PMID- 15809845 TI - Adrenal myelolipoma in a child. AB - A case of adrenal myelolipoma is reported in a 14-year-old girl. Abdominal ultrasound examination revealed a left-sided ovarian cyst 13 cm in diameter and an 8 x 8-cm hyperechoic heterogenous solid mass localized in the right adrenal gland. The ovarian cyst and adrenal mass were removed surgically. Histological examination of the adrenal mass revealed a proliferation of mature adipose tissue with bone marrow-like hematopoietic elements. The ovarian cyst was a simple serous cyst. PMID- 15809846 TI - Asymptomatic gastric duplication in a child: report of a new case and review of the literature. AB - Antral duplication cyst with an antral mucosal lining accounts for 2.2% of all gastric duplications, which represent 4-8% of all alimentary tract duplications. They usually become symptomatic before 2 years of age. We report a 6-year-old child with no previous history of symptoms who was admitted to our hospital because of frequent vomiting and acute abdominal pains for the previous 2 days. Abdominal examination revealed a mass in the epigastric area. PMID- 15809847 TI - Total absence of the small bowel in a premature neonate. AB - We report here an interesting unique case of total loss of small bowel in the absence of associated malrotation or gastroschisis. Total loss of small bowel, acquired in utero in the absence of associated anomalies such as malrotation or gastroschisis, has not been previously reported. Several reports of congenital short bowel exist. However, this uncommon finding is typically associated with malrotation. Babies with this condition often present with functional intestinal obstruction. Several cases in which infarction of gastroschisis and autolysis of the bowel, followed by in utero resolution of the abdominal wall defect, have been reported as a cause of congenital absence of the small bowel. We present here the first report, to our knowledge, of an infant with total absence of the small bowel without gastroschisis or malrotation. PMID- 15809848 TI - Exploratory integration of peanut genetic and physical maps and possible contributions from Arabidopsis. AB - Arachis hypogaea is a widely cultivated crop both as an oilseed and protein source. The genomic analysis of Arachis species hitherto has been limited to the construction of genetic maps; the most comprehensive one contains 370 loci over 2,210 cM in length. However, no attempt has been made to analyze the physical structure of the peanut genome. To investigate the practicality of physical mapping in peanut, we applied a total of 117 oligonucleotide-based probes ("overgos") derived from genetically mapped RFLP probes onto peanut BAC filters containing 182,784 peanut large-insert DNA clones in a multiplex experimental design; 91.5% of the overgos identified at least one BAC clone. In order to gain insights into the potential value of Arabidopsis genome sequence for studies in divergent species with complex genomes such as peanut, we employed 576 Arabidopsis-derived overgos selected on the basis of maximum homology to orthologous sequences in other plant taxa to screen the peanut BAC library. A total of 353 (61.3%) overgos detected at least one peanut BAC clone. This experiment represents the first steps toward the creation of a physical map in peanut and illustrates the potential value of leveraging information from distantly related species such as Arabidopsis for both practical applications such as comparative map-based cloning and shedding light on evolutionary relationships. We also evaluated the possible correlation between functional categories of Arabidopsis overgos and their success rates in hybridization to the peanut BAC library. PMID- 15809849 TI - Coding region single nucleotide polymorphism in the barley low-pI, alpha-amylase gene Amy32b. AB - Barley alpha-amylase variability influences the quality of barley grain in the brewing, feed and food industries. alpha-Amylase proteins are encoded by multigene families in cereals, and this study focused on the barley Amy32b gene. We identified coding region single nucleotide polymorphism (cSNP) and insertion/deletion variation in DNA sequences, which resulted in amino acid substitution and stop codon formation, respectively. The substitution affected the beta1 strand in domain C, whereas the stop codon removed the beta5 strand. Possible effects of these changes on the protein are discussed. A cSNP in the coding region of the Amy32b gene was used as a specific marker to map Amy32b loci on chromosome 7H. PMID- 15809850 TI - SSCP-SNP in pearl millet--a new marker system for comparative genetics. AB - A considerable array of genomic resources are in place in pearl millet, and marker-aided selection is already in use in the public breeding programme at ICRISAT. This paper describes experiments to extend these publicly available resources to a single nucleotide polymorphism (SNP)-based marker system. A new marker system, single-strand conformational polymorphism (SSCP)-SNP, was developed using annotated rice genomic sequences to initially predict the intron exon borders in millet expressed sequence tags (ESTs) and then to design primers that would amplify across the introns. An adequate supply of millet ESTs was available for us to identify 299 homologues of single-copy rice genes in which the intron positions could be precisely predicted. PCR primers were then designed to amplify approximately 500-bp genomic fragments containing introns. Analysis of these fragments on SSCP gels revealed considerable polymorphism. A detailed DNA sequence analysis of variation at four of the SSCP-SNP loci over a panel of eight inbred genotypes showed complex patterns of variation, with about one SNP or indel (insertion-deletion) every 59 bp in the introns, but considerably fewer in the exons. About two-thirds of the variation was derived from SNPs and one-third from indels. Most haplotypes were detected by SSCP. As a marker system, SSCP-SNP has lower development costs than simple sequence repeats (SSRs), because much of the work is in silico, and similar deployment costs and through-put potential. The rates of polymorphism were lower but useable, with a mean PIC of 0.49 relative to 0.72 for SSRs in our eight inbred genotype panel screen. The major advantage of the system is in comparative applications. Syntenic information can be used to target SSCP-SNP markers to specific chromosomal regions or, conversely, SSCP-SNP markers can be used to unravel detailed syntenic relationships in specific parts of the genome. Finally, a preliminary analysis showed that the millet SSCP-SNP primers amplified in other cereals with a success rate of about 50%. There is also considerable potential to promote SSCP-SNP to a COS (conserved orthologous set) marker system for application across species by more specifically designing primers to precisely match the model genome sequence. PMID- 15809851 TI - Stability of QTLs for rice grain dimension and endosperm chalkiness characteristics across eight environments. AB - Rice appearance quality, including traits specifying grain dimension and endosperm chalkiness, represents a major problem in many rice-producing areas of the world. In this study, the genetic basis of six appearance quality traits of milled rice was dissected into quantitative trait loci (QTL) main effects, and the stability of these QTLs was assessed in a population of 66 chromosome segment substitution lines (CSSLs) across eight environments. The CSSLs showed transgressive segregation for many of the traits, and significant correlations were detected among most of the traits. Twenty-two QTLs were identified on eight chromosomes, and numerous QTLs affecting related traits were mapped in the same regions, probably reflecting pleiotropic effects. Nine QTLs, namely qGL-1,qGL-3, qGW-5,qLWR-3, qLWR-5,qPGWC-8, qPGWC-9, qACE-8, and qDEC-8, were consistently detected across the eight environments. The additive main effect and multiplicative interaction (AMMI) analysis showed that genotype (G) x environment (E) interaction was significant for all six traits, with the first three iPCA terms accounting for over 80% of the G x E variance. Both D(I) values and the iPCA1-iPCA2 biplots showed that the CSSLs harboring the nine QTL alleles were more stable than those carrying any of the additional 13 QTL alleles, thereby confirming their environmental stability and pointing to their appropriateness as targets for marker-assisted selection for high-quality rice varieties. PMID- 15809853 TI - [Incidence of transmission of pathogens in intensive care units. Results of the SIR 3 study]. AB - The objective of this study was to determine the incidence of episodes of transmission of nosocomial pathogens and of those pathogens leading to nosocomial infections. Over a period of 18 months all patients from 5 intensive care units (ICUs) who stayed for more than 2 days were included in this study. Surveillance of nosocomial infections was carried out and all isolates of 10 of the most frequent pathogens in ICUs (indicator pathogens) were collected and typed. A total of 28,498 patient days and 431 nosocomial infections were observed (incidence density 15.1 per 1,000 patient days), among them 278 caused by 1 of the selected indicator pathogens. A total of 141 episodes of transmissions were identified, corresponding to an incidence of episodes of transmission of 5.0 per 1,000 patient days and 41 nosocomial infections were transmission-associated, corresponding to 14.5% of all nosocomial infections. The data of this study demonstrate that even in ICUs with average nosocomial infection rates, some nosocomial infections could be avoided. PMID- 15809855 TI - [The future of medical office-based specialists]. PMID- 15809854 TI - [Anesthesia and sleep apnea syndrome]. AB - The perioperative risk for patients with obstructive sleep apnea syndrome and the optimal anaesthesiological management of these patients have not been well elucidated. The prevalence of obstructive sleep apnea with significant symptoms is estimated to be 4% in men and 2% in women. However, in 80-95% of patients this syndrome is not sufficiently diagnosed. Thus identification of patients at risk and a thorough multidisciplinary diagnostic approach are essential for optimal perioperative management. The risk of perioperative complications, like cardiopulmonary compromise, and difficulties in airway management is elevated. The most important aspects of perioperative management include evaluation of intubating conditions, careful search for cardiopulmonary morbidity, permanent control of patient airways, sensible use of anaesthetics, sedatives, and narcotics, and strict monitoring of vital signs. If ambulatory nasal continuous positive airway pressure (CPAP) therapy has been established preoperatively, this should be continued in the perioperative period. Postoperative monitoring should be performed in an intensive care or intermediate care unit. Controlled clinical studies on the best perioperative management of patients with obstructive sleep apnea are urgently required. PMID- 15809856 TI - [Urinary retention with ruptured fornix caused by a maggot. An autoerotic accident]. AB - Foreign bodies in the urinary tract represent a possible cause for genitourinary disease. A huge variety of objects are inserted into the urinary tract for therapeutic, autoerotic, or psychiatric reasons and the symptoms for foreign bodies can be rather unspecific. Dysuria, hematuria, and urinary tract infection can suggest the presence of intravesical foreign bodies. Symptomatic urinary retention is more seldom and the causal foreign bodies are mostly iatrogenically applied. We present a unique case of maggots self-inserted into the urinary tract for autoerotic reasons causing symptomatic urinary retention with fornix rupture. PMID- 15809860 TI - Endovascular occlusion of direct carotid cavernous fistula with detachable balloons: usefulness of 3D angiography. AB - The objective of this study is to show rotational 3D angiography findings and their usefulness in the occlusion of carotid cavernous fistulas (CCFs) using detachable balloons. Five patients with direct CCF were retrospectively reviewed for details of interventional procedures and 2D and 3D angiography findings. Pretherapeutic 2D and 3D angiograms (n=2) were compared to evaluate the size of the fistula and the relative size of the cavernous sinus with respect to the fistula. Postinflation-predetachment (n=3) and postdetachment (n=4) 2D and 3D angiograms were compared in each stage to evaluate the relative location of the balloon to the internal carotid artery (ICA), presence of a pseudoaneurysm, and relative size of the balloon to the fistula. Pretherapeutic 2D and 3D angiograms were equally effective in showing the fistula and relative size of the cavernous sinus. But, 3D angiography with "cut images" at arbitrary viewing angles clearly visualized the 3D relations between the ICA, fistula, and cavernous sinus. Both postinflation-predetachment 2D and 3D images in two patients equally showed a contrast-filled pseudoaneurysm outside the ICA and intraluminal location of the balloon. However, only the 3D images showed no difference in size of the balloon compared with the fistula, which was relevant to traction-induced instability in the remaining one patient. Both postdetachment 2D and 3D angiograms were equal in terms of showing ICA compromise (60%) in one patient and an extraluminal balloon location with complete fistular occlusion in two patients. In the remainder, a small pseudoaneurysm was identified only on 3D images. Three-dimensional angiography is a useful imaging tool for capturing the complex perifistular anatomy in the pretherapeutic stage, and for providing detailed information about the degree of balloon inflation and its location, the presence of a pseudoaneurysm, and the expected traction-induced instability in the predetachment stage. Postinflation-predetachment 3D angiography may therefore offer a chance to correct an erroneous manipulation that would otherwise lead to an incomplete procedural outcome and disastrous balloon migration. PMID- 15809861 TI - Responsive modes of Medicago sativa proline dehydrogenase genes during salt stress and recovery dictate free proline accumulation. AB - Free proline accumulation is an innate response of many plants to osmotic stress. To characterize transcriptional regulation of the key proline cycle enzymes in alfalfa (Medicago sativa), two proline dehydrogenase (MsPDH) genes and a partial sequence of Delta (1) -pyrroline-5-carboxylate dehydrogenase (MsP5CDH) gene were identified and cloned. The two MsPDH genes share a high nucleotide sequence homology and a similar exon/intron structure. Estimation of transcript levels during salt stress and recovery revealed that proline accumulation during stress was linearly correlated with a strong decline in MsPDH transcript levels, while Delta (1) -pyrroline-5-carboxylate synthetase (MsP5CS) and MsP5CDH steady-state transcript levels remained essentially unchanged. MsPDH transcript levels dramatically decreased in a fast, salt concentration-dependent manner. The extent of salt-induced proline accumulation also correlated with salt concentrations. Salt-induced repression of MsPDH1 promoter linked to the GUS reporter gene confirmed that the decline in MsPDH transcript levels was due to less transcription initiation. Contrary to the salt-dependent repression, a rapid induction of MsPDH transcription occurred at a very early stage of the recovery process, independently of earlier salt treatments. Hence our results suggest the existence of two different regulatory modes of MsPDH expression; the repressing mode that quantifies salt concentration in an as yet unknown mechanism and the "rehydration"-enhancing mode that responds to stress relief in a maximal induction of MsPDH transcription. As yet the components of salt sensing as well as those that might interact with MsPDH promoter to reduce transcription are still unknown. PMID- 15809862 TI - Three dimensional confocal and electron microscopy imaging define the dynamics and mechanisms of diploidisation at early stages of barley microspore-derived embryogenesis. AB - In order to determine the timing and mechanisms of the spontaneous diploidisation throughout microspore-derived embryogenesis in barley, we have estimated the ploidy level of individual nuclei within young pro-embryos, from the first androgenetic division up to multinuclear structures still surrounded by the exine. Our methodological approach was based on the measure of the intensity of fluorescence after 4,6-Diamidino-2-phenylindole dihydrochloride staining, nuclear size and number of nucleoli in the confocal microscope. This method avoids the overlapping of the fluorescence signal in multinuclear pro-embryos, which cannot be studied using cytophotometer methods based on other types of fluorescence microscopes. The identification of haploid and diploid nuclei enabled us to determine the timing of diploidisation at early stages throughout androgenetic development. We found that diploidisation is an ongoing process that can start after the first embryogenic division and continues in multinuclear pro-embryos. Reconstruction of 3D-images of entire pro-embryos and the observation of cross and longitudinal sections across stacks of optical sections, together with correlative light and electron microscopy, provided evidences of nuclear fusion as the main mechanism of diploidisation. PMID- 15809863 TI - Comparative analysis of the two-step reaction catalyzed by prokaryotic and eukaryotic phytochelatin synthase by an ion-pair liquid chromatography assay. AB - Genes encoding phytochelatin (PC) synthase have been found in higher plants, fission yeast and worm. Recently, kinetic and mutagenic analyses of recombinant PC synthase have been revealing the molecular mechanisms underlying PC synthesis, however, a conclusive model has not been established. To clarify the mechanism of PC synthase found in eukaryotes, we have compared the two-step reactions catalyzed by the prokaryotic Nostoc PC synthase (NsPCS) and the eukaryotic Arabidopsis PC synthase (AtPCS1). Comparative analysis shows that in the first step of PC synthesis corresponding to the cleavage of gamma-glutamylcysteine (gamma-EC) from glutathione (GSH), free GSH or PCs acts as a donor molecule to supply a gamma-EC unit for elongation of the PC chain, and heavy metal ions are required to carry out the cleavage. Furthermore, functional analyses of various mutants of NsPCS and AtPCS1, selected by comparing the sequences of NsPCS and AtPCS1, indicate that the N-terminal region (residues 1-221) in AtPCS1 is the catalytic domain, and in this region, the Cys(56) residue is associated with the PC synthesis reaction. These results enable us to propose an advanced model of PC synthesis, describing substrate specificity, heavy metal requirement, and the active site in the enzyme. PMID- 15809864 TI - Auxin-cytokinin and auxin-gibberellin interactions during morphogenesis of the compound leaves of pea (Pisum sativum). AB - A number of mutations that alter the form of the compound leaf in pea (Pisum sativum) has proven useful in elucidating the role that auxin might play in pea leaf development. The goals of this study were to determine if auxin application can rescue any of the pea leaf mutants and if gibberellic acid (GA) plays a role in leaf morphogenesis in pea. A tissue culture system was used to determine the effects of various auxins, GA or a GA biosynethesis inhibitor (paclobutrazol) on leaf development. The GA mutant, nana1 (na1) was analyzed. The uni-tac mutant was rescued by auxin and GA and rescue involved both a conversion of the terminal leaflet into a tendril and an addition of a pair of lateral tendrils. This rescue required the presence of cytokinin. The auxins tested varied in their effectiveness, although methyl-IAA worked best. The terminal tendrils of wildtype plantlets grown on paclobutrazol were converted into leaflets, stubs or were aborted. The number of lateral pinna pairs produced was reduced and leaf initiation was impaired. These abnormalities resembled those caused by auxin transport inhibitors and phenocopy the uni mutants. The na1 mutant shared some morphological features with the uni mutants; including, flowering late and producing leaves with fewer lateral pinna pairs. These results show that both auxin and GA play similar and significant roles in pea leaf development. Pea leaf morphogenesis might involve auxin regulation of GA biosynthesis and GA regulation of Uni expression. PMID- 15809865 TI - Interaction between photorespiration and respiration in transgenic potato plants with antisense reduction in glycine decarboxylase. AB - Potato (Solanum tuberosum L. cv. Desiree) plants with an antisense reduction in the P-protein of the glycine decarboxylase complex (GDC) were used to study the interaction between respiration and photorespiration. Mitochondria isolated from transgenic plants had a decreased capacity for glycine oxidation and glycine accumulated in the leaves. Malate consumption increased in leaves of GDC deficient plants and the capacity for malate and NADH oxidation increased in isolated mitochondria. A lower level of alternative oxidase protein and decreased partitioning of electrons to the alternative pathway was found in these plants. The adenylate status was altered in protoplasts from transgenic plants, most notably the chloroplastic ATP/ADP ratio increased. The lower capacity for photorespiration in leaves of GDC deficient plants was compensated for by increased respiratory decarboxylations in the light. This is interpreted as a decreased light suppression of the tricarboxylic acid cycle in GDC deficient plants in comparison to wild-type plants. The results support the view that respiratory decarboxylations in the light are restricted at the level of the pyruvate dehydrogenase complex and/or isocitrate dehydrogenase and that this effect is likely to be mediated by mitochondrial photorespiratory products. PMID- 15809866 TI - Reorganized actin filaments anchor chloroplasts along the anticlinal walls of Vallisneria epidermal cells under high-intensity blue light. AB - In epidermal cells of the aquatic angiosperm Vallisneria gigantea Graebner, high intensity blue light (BL) induces the avoidance response of chloroplasts. We examined simultaneous BL-induced changes in the configuration of actin filaments in the cytoplasmic layers that face the outer periclinal wall (P side) and the anticlinal wall (A side). The results clearly showed that dynamic reorganization of the actin cytoskeleton occurs on both sides. Upon BL irradiation, thick, long bundles of actin filaments appeared, concomitant with the directed migration of chloroplasts from the P side to the A side. After 15-20 min of BL irradiation, fine actin bundles on only the A side appeared to associate with chloroplasts that had migrated from the P side. To examine the role of the fine actin bundles, we evaluated the anchorage of chloroplasts by centrifuging living cells. Upon BL irradiation, the resistance of chloroplasts on both the P and A sides to the centrifugal force decreased remarkably. After 20 min of BL irradiation, the resistance of chloroplasts on the A side increased again, but chloroplasts on the P side could still be displaced. The BL-induced recovery of resistance of chloroplasts on the A side was sensitive to photosynthesis inhibitors but insensitive to an inhibitor of flavoproteins. The photosynthesis inhibitors also prevented the fine actin bundles from appearing on the A side under BL irradiation. These results strongly suggest that the BL-induced avoidance response of chloroplasts includes photosynthesis-dependent and actin-dependent anchorage of chloroplasts on the A side of epidermal cells. PMID- 15809868 TI - Genome-wide mRNA profiling in glucose starved Bacillus subtilis cells. AB - In this study global changes in gene expression were monitored in Bacillus subtilis cells entering stationary growth phase owing to starvation for glucose. Gene expression was analysed in growing and starving cells at different time points by full-genome mRNA profiling using DNA macroarrays. During the transition to stationary phase we observed extensive reprogramming of gene expression, with approximately 1,000 genes being strongly repressed and approximately 900 strongly up-regulated in a time-dependent manner. The genes involved in the response to glucose starvation can be assigned to two main classes: (i) general stress/starvation genes which respond to various stress or starvation stimuli, and (ii) genes that respond specifically to starvation for glucose. The first class includes members of the sigma(B)-dependent general stress regulon, as well as 90 vegetative genes, which are strongly down regulated in the course of the stringent response. Among the genes in the second class, we observed a decrease in the expression of genes encoding proteins required for glucose uptake, glycolysis and the tricarboxylic acid cycle. Conversely, many carbohydrate utilisation systems that depend on phosphotransferase systems (PTS) or ABC transporters were activated. The expression of genes required for utilisation or generation of acetate indicates that acetate constitutes an important energy source for B. subtilis during periods of glucose starvation. Finally, genome wide mRNA profiling data can be used to predict new metabolic pathways in B. subtilis. Thus, our data suggest that glucose-starved cells are able to degrade branched chain fatty acids to pyruvate and succinate via propionyl-CoA using the methylcitrate pathway. This pathway appears to link lipid degradation to gluconeogenesis in glucose-starved cells. PMID- 15809869 TI - Phosphorus source alters host plant response to ectomycorrhizal diversity. AB - We examined the influence of phosphorus source and availability on host plant (Pinus rigida) response to ectomycorrhizal diversity under contrasting P conditions. An ectomycorrhizal richness gradient was established with equimolar P supplied as either inorganic phosphate or organic inositol hexaphosphate. We measured growth and N and P uptake of individual P. rigida seedlings inoculated with one, two, or four species of ectomycorrhizal fungi simultaneously and without mycorrhizas in axenic culture. Whereas colonization of P. rigida by individual species of ectomycorrhizal fungi decreased with increasing fungal richness, colonization of all species combined increased. Plant biomass and N content increased across the ectomycorrhizal richness gradient in the organic but not the inorganic P treatment. Plants grown under organic P conditions had higher N concentration than those grown under inorganic P conditions, but there was no effect of richness. Phosphorus content of plants grown in the organic P treatment increased with increasing ectomycorrhizal richness, but there was no response in the inorganic P treatment. Phosphorus concentration was higher in plants grown at the four-species richness level in the organic P treatment, but there was no effect of diversity under inorganic P conditions. Overall, few ectomycorrhizal composition effects were found on plant growth or nutrient status. Phosphatase activities of individual ectomycorrhizal fungi differed under organic P conditions, but there was no difference in total root system phosphatase expression between the inorganic or organic P treatments or across richness levels. Our results provide evidence that plant response to ectomycorrhizal diversity is dependent on the source and availability of P. PMID- 15809870 TI - Distribution of dominant arbuscular mycorrhizal fungi among five plant species in undisturbed vegetation of a coastal grassland. AB - Most plant species in mixed grassland vegetation are colonized by arbuscular mycorrhizal (AM) fungi. Previous studies have reported differences in host preferences among AM fungi, although the fungi are known to lack host specificity. In the present study, the distribution of phylogenetic groups of AM fungi belonging to a clade of Glomus species was studied in five plant species from a coastal grassland in Denmark. The occurrence of the fungi was determined by PCR analyses of fungal large subunit ribosomal DNA sequences amplified from root fragments using a specific primer set. The results showed that the dominant Glomus species were able to colonize all the studied plant species, supporting the view that the AM fungi represent a large underground interconnecting mycelial network. PMID- 15809871 TI - Intra- and interobserver variability during in vitro validation of two novel methods for intra-abdominal pressure monitoring. AB - OBJECTIVE: Intra-abdominal pressure (IAP) measurement techniques vary in automaticity and reproducibility. This study evaluated the intra- and interobserver variability of two new IAP measurement techniques. METHODS: A one half open, 30-l container was used with two IAP catheters (Foley Manometer, balloon-tipped catheter) contained in a 100-ml infusion bag. To simulate intra abdominal hypertension the container was filled with water using 5-cm increments (0-25 cmH2O). Pressure was determined by observers using the Foley Manometer and simultaneously recorded using an IAP monitor. Observers were blinded to the reference levels. RESULTS: Fifteen observers conducted three pressure readings at each of the six pressure levels with the Foley technique, giving 270 readings. These were paired with the automated monitor readings and the height of the water column. The intra- and interobserver coefficients of variation were low for both methods. The Spearman correlation coefficient was higher than 0.9 for all paired measurements and Bland-Altman analysis comparing the reference H2O column to both measurement techniques showed a very good agreement at all pressure intervals (bias 0.1+/-0.6 cmH2O) and a consistent, low underestimation of the reference water column pressure by both techniques. CONCLUSIONS: Both the Foley Manometer and the IAP monitor are reliable and reproducible methods to measure IAP in this in vitro model. The coefficient of variation for each technique is low and decreases with increasing IAP, the monitor giving more reproducible results than the Foley Manometer. PMID- 15809872 TI - Percutaneous treatment of haemophilic digital pseudo tumours. AB - In young patients with haemophilia, blood cysts in the finger bones may grow rapidly and imitate a malignant tumour (pseudo tumour). The condition must initially be treated with administration of factor VIII-IX. If this does not control the growth, surgical treatment is indicated. We have advised a percutaneous treatment with evacuation of the blood and injection of the cavity with fibrin seal. PMID- 15809873 TI - Comparison between mobile-bearing and fixed-bearing knees in bilateral total knee replacements. AB - The purpose of this study was to compare mid-term results of mobile-bearing and fixed-bearing in bilateral total knee arthroplasty (TKA). Twenty-two patients underwent bilateral TKA with a mobile-bearing prosthesis (Rotaglide, Corin, UK) on one side and a fixed-bearing prosthesis (NexGen-CR, Zimmer, USA) on the other. There were 21 female patients, and in 18 patients, the diagnosis was rheumatoid arthritis. The average age was 59.6 (35-78) years. In all procedures, the posterior cruciate ligament was retained and patella re-surfaced. The average follow-up in the mobile-bearing group was 98 (79-107) months and 96 (79-107) months in the fixed-bearing group. At the final follow-up, the knee score was 91.8 points and 91.1 points, respectively, and the function score 65.5 points. The range of motion was similar in the two groups (1.1-106.9 degrees; 0.4-106.9 degrees). Five patients favoured the fixed-bearing prosthesis, but 16 found no difference. In patients with bilateral TKA, there was no difference in the short term result between mobile-bearing and fixed-bearing prostheses. PMID- 15809874 TI - Structural allograft and primary press-fit cup for severe acetabular deficiency. A minimum 6-year follow-up study. AB - Between October 1992 and December 1996, 23 patients with pelvic bone stock deficiency involving major columns underwent revision surgery with a cementless press-fit cup and a structural bone graft. Twenty cases were followed up for a minimum of 6 (average 7.6, range 6-11) years. Three cups were revised: one for aseptic loosening, one for septic loosening, and one for recurrent dislocation. At latest follow-up, the average Merle d'Aubigne hip score improved from 10.9 to 16.2; four hips were rated excellent, seven very good, three good, two fair, and one poor. All cups were stable; the grafts were integrated and anatomy was restored. The Kaplan-Meier cumulative probability of not having revision for loosening at 11 years, predicted a survival rate of 84.4%. We are confident that these results are satisfactory for a very demanding procedure. PMID- 15809875 TI - Differential gene expression in filamentous cells of Ustilago maydis. AB - When fungi interact with plants as pathogens or as symbionts, there are often changes in fungal cell morphology and nuclear state. This study establishes the use of cDNA microarrays to detect gene expression changes in Ustilago maydis cells that differ in structure and nuclear content. Categorizing differentially expressed genes on the basis of function indicated that U. maydis cell types vary most in the expression of genes related to metabolism. We also observed that more genes are up-regulated in the filamentous dikaryon than in the filamentous diploid, relative to non-pathogenic budding cells. Our comparison of pathogenic development indicated that the dikaryon is more virulent than the diploid. Other identified expression patterns suggest a cell-specific difference in nutrient acquisition, cell metabolism and signal transduction. The relevance of gene expression change to cell type biology is discussed. PMID- 15809876 TI - Saccharomyces cerevisiaeTSC11/AVO3 participates in regulating cell integrity and functionally interacts with components of the Tor2 complex. AB - Saccharomyces cerevisiae TSC11/AVO3 is an essential gene encoding one component of TORC2, a multi-protein complex of yeast Tor2p that also contains Lst8p, Avo1p, and Avo2p. Despite the proven physical association among TORC2 components, little is known about the functional linkage or cellular pathways these proteins act in. Here, we present genetic data linking the function of TSC11 to the regulation of cell integrity. Mutants carrying temperature-sensitive (ts) alleles in different regions of TSC11 displayed cell wall defects, evidenced by characteristic osmotic stabilizer-remediable cell lysis, susceptibility to trypan blue staining, and sensitivity to cell wall-digesting enzymes. Dosage suppression analysis identified different groups of genes in rescuing phenotypes of different tsc11(ts) mutants. AVO1 suppressed one class of mutants, whereas active PKC1, AVO2, and SLM1 partially rescued another. Our findings demonstrate functional connections among TORC2 components and we speculate that Tsc11p exerts its function via a Pkc1p-independent mechanism mediated through Avo1p, and a Pkc1p dependent mechanism mediated through Avo2p and Slm1p. PMID- 15809877 TI - Phase I study of the multidrug resistance inhibitor zosuquidar administered in combination with vinorelbine in patients with advanced solid tumours. AB - BACKGROUND: Zosuquidar (LY335979) is an oral P-glycoprotein modulator. This phase I study was designed to determine the maximum tolerated dose (MTD) of zosuquidar in combination with vinorelbine. The effects of zosuquidar on vinorelbine pharmacokinetics were also examined. DESIGN: Patients with advanced solid tumours were treated with escalating doses of zosuquidar administered every 8-12 h on days 7-9 and 14-16 during cycle 1 then days 0-2, 7-9, and 14-16 from cycle 2 onwards, with vinorelbine 22.5-30 mg/m2 IV on days 1, 8 and 15 every 28 days. RESULTS: Of 21 patients registered, 19 were treated at four dose levels (zosuquidar 100-300 mg/m2). Two patients had prolonged and febrile neutropenia at the second dose level resulting in a reduction of the dose of vinorelbine in subsequent dose levels. There was another patient with dose-limiting febrile neutropenia at dose level four which resulted in the expansion of the dose level three. Eight patients had stable disease and no objective responses were seen. Vinorelbine pharmacokinetic studies showed reduced clearance when given with zosuquidar. CONCLUSIONS: The MTD was zosuquidar 300 mg/m2 orally every 12 h for 3 days weekly for 3 weeks with vinorelbine 22.5 mg/m2 IV weekly for 3 weeks every 28 days. Zosuquidar may inhibit vinorelbine clearance to a modest degree. PMID- 15809878 TI - Phase I clinical and pharmacokinetic trial of docetaxel and irinotecan administered on a weekly schedule. AB - BACKGROUND: Docetaxel and irinotecan are synergistic agents with a broad spectrum of activity but overlapping myelosuppression. The study was designed to maintain dose intensity while limiting myelosuppression. The objectives of this study were to determine the maximal tolerated dose (MTD) of the combination of docetaxel and irinotecan administered weekly for four consecutive weeks every 42 days, to describe toxicities of this regimen, and to perform a pharmacokinetic analysis to evaluate changes in drug disposition as a function of dose as well as repeated dosing. METHODS: Adult patients with advanced solid tumors were treated with docetaxel followed by irinotecan. Doses of 30/50, 35/50, 35/66, 30/57, 30/65, 30/80 mg/m(2), respectively, were studied. Pharmacokinetics of docetaxel, irinotecan and SN-38 in plasma were determined on days 1 and 22 by a high performance liquid chromatography (HPLC) assay. RESULTS: A total of 35 patients were treated. The MTD was docetaxel 30 mg/m(2) plus irinotecan 65 mg/m(2). Diarrhea was the dose-limiting toxicity; myelosuppression and other non hematological toxicities were uncommon and mild. There were no significant differences in pharmacokinetic parameters between day 1 and day 22 (n=20). Five objective responses (breast, stomach and unknown primary) were observed among 30 evaluable patients. In addition, eight patients achieved stable disease. CONCLUSIONS: The combination of weekly docetaxel and irinotecan is a well tolerated regimen and should be explored in phase II trials. This schedule maintains dose intensity and has limited myelosuppression. PMID- 15809879 TI - [Tracheobronchial ruptures: classification and management]. AB - BACKGROUND: Tracheobronchial ruptures are rare surgical emergencies with significant mortality. We present management and outcome of such ruptures treated at the University of Leipzig in Germany and propose a novel therapeutic algorithm a new classification system for stratifying treatment of patients with tracheobronchial ruptures. METHODS: We retrospectively studied 24 patients 19 to 88 years old who were treated in our institution for tracheobronchial injury. RESULTS: Eighty-seven percent of the injuries were caused iatrogenically. Fifty four percent were type I injuries (isolated tracheal lesions), 38% type II (involvement of carina or main stem bronchi), and 8% type III (distal lesions of lobar or segmental bronchi). Seventy-five percent of the patients were operated via right-sided dorsolateral thoracotomy. In four (22%), insufficiency of the tracheal closure occurred, with mediastinitis possibly being a significant risk factor for this event (P<0.001). In surgically treated patients, rupture-related and overall mortality were 5.5% and 28%, respectively, whereas in medically treated patients, mortality was 33%. CONCLUSION: The proposed classification of tracheobronchial injuries enables stratifying the treatment of patients with tracheobronchial ruptures. Type I lesions can be surgically closed either by a right-sided thoracotomy or transcervical-transtracheal approach. In contrast, surgical management of type II and III injuries always requires thoracotomy. PMID- 15809880 TI - Prognostic value of cyclin D1 overexpression in correlation with pRb and p53 status in non-small cell lung cancer (NSCLC). AB - PURPOSE: The aim of this study was to assess the impact of cyclin D1 overexpression (considered separately or jointly with previously assessed p53 and pRb statuses) on survival in a group of 111 surgically treated non-small cell lung cancer patients (NSCLC). METHODS: Cyclin D1 accumulation was assessed immunohistochemically, with the use of monoclonal antibody (DCS-6, DakoCytomation) and the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique. RESULTS: Overexpression of cyclin D1 was found in 55 samples (49%), whereas the altered phenotypes cyclin D1+/p53+ or cyclin D1+/pRb- were found in 23 (22%) and 9 samples (9%), respectively. Statistical analysis was performed for different cut-off values and the only significant differences were found if samples with some expression of each protein were considered positive. There was no relationship between cyclin D1 overexpression and major clinicopathological factors, including p53 expression; however, there was a direct correlation between cyclin D1 and pRb protein expression (p=0.007). Cyclin D1 accumulation did not influence patients' survival. Of all possible cyclin D1/p53, cyclin D1/pRb and cyclin D1/p53/pRb phenotypes, patients with cyclin D1-/p53+ phenotype had shortened overall survival compared to other patients (p=0.027, HR=1.8). In the multivariate analysis, the only variable associated with shortened overall and disease-free survival was the stage of disease (p<0.001). CONCLUSIONS: These results suggest the lack of prognostic value of cyclin D1 overexpression in NSCLC patients. PMID- 15809881 TI - The fusion allele of the FUT2 (secretor type alpha(1,2)-fucosyltransferase) gene at a high frequency and a new se385 allele in a Korean population. AB - The fusion gene (se(fus)), a nonfunctional allele of the FUT2 [secretor type alpha(1,2)-fucosyltransferase] gene, was found in Japanese populations with high frequencies (4.8-7.9%). In a study on a Korean population, se(fus) was found at a very low frequency (0.6%), but it has not yet been revealed in any other ethnic population. The aim of the present study was to investigate FUT2 gene polymorphisms in a Korean population and to evaluate their implications in secretor expression in saliva. We investigated the frequency of the FUT2 alleles in a Korean population via polymerase chain reaction with confronting two-pair primers (PCR-CTPP) and restriction fragment length polymorphism (PCR-RFLP). Blood samples were collected from 348 random donors in Bundang Koreans. From a total of 696 alleles examined, the frequency of the se(fus) allele in the Korean population was 10.8%. In addition, the new se385 allele was found in about 7.2% of the subjects, an unusually frequent occurrence compared to any other population investigated so far. The null alleles of the FUT2 gene are another example of rare alleles occurring with unexpectedly high frequencies in distinct geographic regions or populations. PMID- 15809882 TI - Evaluation of potential risk factors for early infectious complications after autologous peripheral blood stem cell transplantation in patients with lymphoproliferative diseases. AB - A number of risk factors for the occurrence of neutropaenic fever after haematopoietic stem cell transplantation (HSCT) have been proposed. We were interested in whether these factors remain valid for several early infection related outcomes when applied to a homogeneous group of patients in uni- and multivariate analyses. Therefore, we analysed 144 consecutive patients with lymphoproliferative disorders receiving autologous peripheral blood HSCT. Variables tested as potential risk factors for the occurrence of fever, documented infection (DI), microbiologically documented infection (MDI) or failure of first-line antimicrobial therapy were sex, conditioning regimen, prolonged neutropaenia, low number of CD34+ cells transplanted, purging, lack of selective gut decontamination, higher age and increased body mass index. In uni- and multivariate analyses, conditioning including total body irradiation was the only risk factor for the occurrence of fever, and neutropaenia >or=10 days was the only factor associated with failure of first-line antimicrobial therapy. None of the variables tested was associated with an increased risk for DI or MDI. This analysis suggests that a number of previously proposed risk factors actually are of minor clinical relevance for early infections in the majority of patients receiving autologous HSCT. PMID- 15809883 TI - Blood mononuclear cells and platelets have abnormal fatty acid composition in homozygous sickle cell disease. AB - Leukocyte adhesion to vascular endothelium contributes to vaso-occlusion and widespread organ damage in sickle cell disease (SCD). Previously, we found high expression of the adhesion molecules alphaMbeta2 integrin and L-selectin in HbSS individuals with severe disease. Since membrane n-6 and n-3 polyunsaturated fatty acids modulate cell adhesion, inflammation, aggregation and vascular tone, we investigated the fatty acid composition of mononuclear cells (MNC) and platelets of HbSS patients in steady state (n=28) and racially matched, healthy HbAA controls with similar age and sex distribution living in the same environment (n=13). MNC phospholipids of the patients had lower levels of docosahexaenoic acid (DHA, p<0.01) and increased arachidonic acid (AA, p<0.005) relative to HbAA controls. Similarly, platelets from HbSS patients had less eicosapentaenoic acid (EPA, p<0.05) and more AA (p<0.05) in choline phosphoglycerides (CPG), with reduced DHA (p<0.05) in ethanolamine phosphoglycerides. Platelet CPG had lower DHA levels in SCD patients with complications compared to those without (p<0.05). Reduced cell content of EPA and DHA relative to AA favours the production of aggregatory and proinflammatory eicosanoids that activate leukocytes and platelets. This facilitates inflammation, leukocyte adhesion, platelet aggregation and vaso-occlusion in SCD. PMID- 15809884 TI - Cyclosporin A in myelodysplastic syndrome: a preliminary report. AB - Therapeutic approaches are not well established in patients with myelodysplastic syndrome (MDS). We evaluated response to cyclosporin A (CyA) in 19 cases with MDS who were enrolled for the study [13 refractory anemia (RA), 5 refractory anemia with excess of blasts (RAEB), and 1 refractory anemia with ringed sideroblasts (RARS)]. Bone marrow was normocellular in ten, hypercellular in five, and hypocellular in four cases. Fifteen patients were transfusion dependent and the rest were not transfusion dependent but with a hemoglobin range of 6.4-8.8 g% with a mean of 7.4 g%. CyA was given at a dose of 3-5 mg/kg per day. A major response was observed in seven patients with RA, which was sustained on follow up. Four cases of RA showed minor response and two cases of RA did not respond to CyA therapy. A minor response was also seen in one RAEB and one RARS case, while one RAEB case that initially showed a major response relapsed on therapy. The first effect of therapy was evident after a mean period of 2.5 months. A rise in platelets and leukocyte count was seen in three and two cases, respectively. One case developed renal failure on therapy and later died of septicemia. Response to CyA was independent of bone marrow cellularity. CyA could be an effective mode of therapy in patients with MDS especially those having RA. PMID- 15809885 TI - Deferiprone as an oral iron chelator in sickle cell disease. AB - Iron overload is not uncommon in sickle cell disease (SCD) and requires regular chelation therapy in several instances. The present study evaluates the effect of deferiprone in 15 adult patients with SCD (ten beta(s)/beta(0)thalassemia and five beta(s)/beta(s)) and iron overload. Deferiprone was given at a dose of 75 mg/kg daily for 12 months. The evaluation considered pre- and post-treatment values of serum ferritin, urinary iron excretion, and T2 values of liver and heart obtained by magnetic resonance imaging (MRI). Eleven patients had a liver biopsy prior to starting therapy to evaluate iron concentration (LIC). Twelve patients completed the study with satisfactory compliance. In ten of them (83.3%) the serum ferritin levels decreased significantly at the end of the trial; in eight patients (66.6%) the reduction of serum ferritin was accompanied by a significant increase of their liver T2 values. All patients had a significant increase of urinary iron excretion in response to the drug. Ferritin levels and liver T2 values correlated with liver iron concentration; on the contrary, ferritin levels and liver T2 values failed to show any correlation with heart T2 values. Heart T2 values did not also show any correlation with left ventricular ejection fraction. Deferiprone was well tolerated and did not cause any significant adverse effects. These results suggest that deferiprone may effectively decrease the iron deposition in patients with SCD; moreover, T2 MRI proves to be a reliable and rapid, noninvasive method for assessing the liver iron load in patients with SCD. PMID- 15809886 TI - Mercury and trace element fractionation in Almaden soils by application of different sequential extraction procedures. AB - A comparative evaluation of the mercury distribution in a soil sample from Almaden (Spain) has been performed by applying three different sequential extraction procedures, namely, modified BCR (three steps in sequence), Di Giulio Ryan (four steps in sequence), and a specific SEP developed at CIEMAT (six steps in sequence). There were important differences in the mercury extraction results obtained by the three procedures according to the reagents applied and the sequence of their application. These findings highlight the difficulty of setting a universal SEP to obtain information on metal fractions of different mobility for any soil sample, as well as the requirement for knowledge about the mineralogical and chemical characteristics of the samples. The specific six-step CIEMAT sequential extraction procedure was applied to a soil profile (Ap, Ah, Bt1, and Bt2 horizons). The distribution of mercury and major, minor, and trace elements in the different fractions were determined. The results indicate that mercury is mainly released with 6 M HCl. The strong association of mercury with crystalline iron oxyhydroxides, present in all the horizons of the profile, and/or the solubility of some mercury compounds in such acid can explain this fact. Minor mercury is found in the fraction assigned to oxidizable matter and in the final insoluble residue (cinnabar). PMID- 15809887 TI - Furofuran lignans from a callus culture of Cichorium intybus. AB - Three new and one known furofuran lignans--syringaresinol derivatives--along with the known phenylpropanoids cichoriin and syringin were isolated from a callus tissue of Cichorium intybus. The compounds were characterised by spectral methods. This is the first report on the presence of furofuran lignans in Cichorium species. PMID- 15809888 TI - Abiotic metal stress enhances diosgenin yield in Dioscorea bulbifera L. cultures. AB - Lower concentrations of CuSO(4) (25-75 microM) in the MS medium supplemented with 0.1 mg l(-1) IAA+5.0 mg l(-1) Kn+500 mg l(-1) CH+10 mg l(-1) Cyst hyd enhanced the growth of regenerants of Dioscorea bulbifera L. CuSO(4) (75 microM) induced an appreciable diosgenin yield in the regenerants compared to those obtained on media without Cu. The presence of Cu thus seems to stimulate diosgenin production. The regenerants also differentiated bulbils on lower concentrations of Cu. At CuSO(4) (100 microM), however, cultures showed poor growth as well as a low diosgenin yield. Increased proline and protein contents were recorded in cultures grown on Cu-enriched media. PMID- 15809889 TI - Can E-selectin be a reliable marker of inflammation in lumbar disc disease? AB - The cause of sciatica and low back pain associating with lumbar disc herniation has not been clearly identified until now. Inflammation has been shown to occur via immunohistochemical and biochemical methods in herniated disc tissues. The important prognostic role of E-selectin has recently been substantiated by other studies in early rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). The important role of adhesion molecules in the initiation and progression of the inflammatory response is well known for infectious diseases and autoimmune disorders. In our study, we aimed to show the role of E-selectin as an inflammatory marker and the correlation of inflammation with straight-leg raise (SLR) test findings and subtype of disc herniation. We found that the cases with positive SLR test had higher rates of immunostaining with E-selectin. This led us to think that E-selectin might play an important role in the activity status of the disease, meaning patients with more limited movement capacity might benefit from E-selectin antagonist therapy. Among the many studies performed to identify the relationship between the inflammation markers and activity of lumbar disc herniation, this is the first investigation held with E-selectin. PMID- 15809890 TI - The cervical spine and its relation to anterior plate-screw fixation: a quantitative study. AB - Multilevel discectomy and inter-vertebral body fusion combined with anterior plate-screw fixation is the common procedure in cervical spine surgery. The correct placement of the screws is an important factor for the outcome of these operations. Yet no systematic approach has been undertaken to optimize the geometry of the fixation-plates regarding the position of the screw-perforations. In this study MRI scans of 50 consecutive patients were analyzed regarding the height of each segment (C3-C7), the anterior-posterior diameter of the vertebral body and the distance between the vertebral arteries. Based on this data we developed "Standard Spine Models". Using these models we designed two plates each for single and two-level surgery, and three plates each for three- and four-level surgery. These ten plates do fit the cervical spines of all 50 patients examined in this study. With these plates the screw-perforations could be positioned efficiently over the bodies of the concerned vertebrae. This should facilitate the selection of a plate and the positioning of the screws. Thus the surgeons might save time and the screws might be positioned more exactly and entirely in the vertebral bodies, ensuring a secure fixation. PMID- 15809893 TI - Amplified HIV transmission and new approaches to HIV prevention. PMID- 15809894 TI - The Effect of early treatment on the latent reservoir of HIV-1. PMID- 15809895 TI - Pharmacogenomics and HIV therapeutics. PMID- 15809896 TI - Airborne severe acute respiratory syndrome coronavirus and its implications. PMID- 15809897 TI - Rates of HIV-1 transmission per coital act, by stage of HIV-1 infection, in Rakai, Uganda. AB - BACKGROUND: We estimated rates of human immunodeficiency virus (HIV)-1 transmission per coital act in HIV-discordant couples by stage of infection in the index partner. METHODS: We retrospectively identified 235 monogamous, HIV discordant couples in a Ugandan population-based cohort. HIV transmission within pairs was confirmed by sequence analysis. Rates of transmission per coital act were estimated by the index partner's stage of infection (recent seroconversion or prevalent or late-stage infection). The adjusted rate ratio of transmission per coital act was estimated by multivariate Poisson regression. RESULTS: The average rate of HIV transmission was 0.0082/coital act (95% confidence interval [CI], 0.0039-0.0150) within approximately 2.5 months after seroconversion of the index partner; 0.0015/coital act within 6-15 months after seroconversion of the index partner (95% CI, 0.0002-0.0055); 0.0007/coital act (95% CI, 0.0005-0.0010) among HIV-prevalent index partners; and 0.0028/coital act (95% CI, 0.0015-0.0041) 6-25 months before the death of the index partner. In adjusted models, early- and late-stage infection, higher HIV load, genital ulcer disease, and younger age of the index partner were significantly associated with higher rates of transmission. CONCLUSIONS: The rate of HIV transmission per coital act was highest during early-stage infection. This has implications for HIV prevention and for projecting the effects of antiretroviral treatment on HIV transmission. PMID- 15809898 TI - Effect of treatment, during primary infection, on establishment and clearance of cellular reservoirs of HIV-1. AB - Patients in whom virologic suppression is achieved with highly active antiretroviral therapy (HAART) retain long-lived cellular reservoirs of human immunodeficiency virus type 1 (HIV-1); this retention is an obstacle to sustained control of infection. To assess the impact that initiating treatment during primary HIV-1 infection has on this cell population, we analyzed the decay kinetics of HIV-1 DNA and of infectivity associated with cells activated ex vivo in 27 patients who initiated therapy before or <6 months after seroconversion and in whom viremia was suppressed to <50 copies/mL. The clearance rates of cellular reservoirs could not be distinguished by these techniques (median half-life, 20 weeks) during the first year of HAART. The clearance of HIV-1 DNA slowed significantly during the subsequent 3 years of treatment (median half-life, 70 weeks), consistent with heterogeneous cellular reservoirs being present. Total cell-associated infectivity (CAI) after 1 year of treatment was undetectable (<0.07 infectious units/million cells [IUPM]) in most patients initiating treatment during primary infection either before (9/9) or <6 months after (6/8) seroconversion. In contrast, all 17 control patients who initiated HAART during chronic infection retained detectable CAI after 3-6 years of treatment (median reservoir size, 1.1 IUPM; P<.0005). These results suggest that treatment <6 months after seroconversion may facilitate long-term control of cellular reservoirs that maintain HIV-1 infection during treatment. PMID- 15809899 TI - Modeling the influence of APOC3, APOE, and TNF polymorphisms on the risk of antiretroviral therapy-associated lipid disorders. AB - BACKGROUND: Single-nucleotide polymorphisms in genes involved in lipoprotein and adipocyte metabolism may explain why dyslipidemia and lipoatrophy occur in some but not all antiretroviral therapy (ART)-treated individuals. METHODS: We evaluated the contribution of APOC3 -482C-->T, -455T-->C, and 3238C-->G; epsilon 2 and epsilon 4 alleles of APOE; and TNF -238G-->A to dyslipidemia and lipoatrophy by longitudinally modeling >2600 lipid determinations and 2328 lipoatrophy assessments in 329 ART-treated patients during a median follow-up period of 3.4 years. RESULTS: In human immunodeficiency virus (HIV)-infected individuals, the effects of variant alleles of APOE on plasma cholesterol and triglyceride levels and of APOC3 on plasma triglyceride levels were comparable to those reported in the general population. However, when treated with ritonavir, individuals with unfavorable genotypes of APOC3 and [corrected] APOE were at risk of extreme hypertriglyceridemia. They had median plasma triglyceride levels of 7.33 mmol/L, compared with 3.08 mmol/L in the absence of ART. The net effect of the APOE*APOC3*ritonavir interaction was an increase in plasma triglyceride levels of 2.23 mmol/L. No association between TNF -238G-->A and lipoatrophy was observed. CONCLUSIONS: Variant alleles of APOE and APOC3 contribute to an unfavorable lipid profile in patients with HIV. Interactions between genotypes and ART can lead to severe hyperlipidemia. Genetic analysis may identify patients at high risk for severe ritonavir-associated hypertriglyceridemia. PMID- 15809900 TI - Cross-reactivity of anti-HIV-1 T cell immune responses among the major HIV-1 clades in HIV-1-positive individuals from 4 continents. AB - BACKGROUND: The genetic diversity of human immunodeficiency virus type 1 (HIV-1) raises the question of whether vaccines that include a component to elicit antiviral T cell immunity based on a single viral genetic clade could provide cellular immune protection against divergent HIV-1 clades. Therefore, we quantified the cross-clade reactivity, among unvaccinated individuals, of anti HIV-1 T cell responses to the infecting HIV-1 clade relative to other major circulating clades. METHODS: Cellular immune responses to HIV-1 clades A, B, and C were compared by standardized interferon- gamma enzyme-linked immunospot assays among 250 unvaccinated individuals, infected with diverse HIV-1 clades, from Brazil, Malawi, South Africa, Thailand, and the United States. Cross-clade reactivity was evaluated by use of the ratio of responses to heterologous versus homologous (infecting) clades of HIV-1. RESULTS: Cellular immune responses were predominantly focused on viral Gag and Nef proteins. Cross-clade reactivity of cellular immune responses to HIV-1 clade A, B, and C proteins was substantial for Nef proteins (ratio, 0.97 [95% confidence interval, 0.89-1.05]) and lower for Gag proteins (ratio, 0.67 [95% confidence interval, 0.62-0.73]). The difference in cross-clade reactivity to Nef and Gag proteins was significant (P<.0001). CONCLUSIONS: Cross-clade reactivity of cellular immune responses can be substantial but varies by viral protein. PMID- 15809901 TI - Response to hepatitis B vaccine in HIV-1-positive subjects who test positive for isolated antibody to hepatitis B core antigen: implications for hepatitis B vaccine strategies. AB - BACKGROUND: Whether human immunodeficiency virus type 1 (HIV-1)-positive subjects who test positive for isolated antibody to hepatitis B core antigen (anti-HBc) should be vaccinated with hepatitis B vaccine is not certain. Development of an anamnestic response after vaccination would suggest previous hepatitis B virus (HBV) infection, in which case vaccination is not necessary. METHODS: Sixty-nine HIV-1-positive subjects who tested negative for hepatitis B surface antigen (HBsAg) and antibody to HBsAg (anti-HBs) received vaccination with standard hepatitis B vaccine. Twenty-nine subjects (42%) tested positive for anti-HBc, and 40 (58%) tested negative for anti-HBc. An anamnestic response was defined as an anti-HBs titer of >or=10 IU/L within 4 weeks of the first vaccination. RESULTS: The overall anamnestic response rate was 16% and was not significantly different between subjects who tested positive for anti-HBc (24%) and those who tested negative for anti-HBc (10%) before vaccination (P=.18). Approximately 50% of subjects who tested positive for anti-HBc also tested positive for antibody to hepatitis Be antigen (anti-HBe). The anamnestic response rate was higher in subjects who tested positive for both anti-HBc and anti-HBe (43%) than in subjects who tested positive for anti-HBc but negative for anti-HBe (7%) (P=.035). After a complete series of vaccinations, HIV-1/hepatitis C virus (HCV) coinfected subjects were less likely to achieve high anti-HBs titers than were subjects infected with HIV-1 alone. CONCLUSIONS: After hepatitis B vaccination, the anamnestic response rate in HIV-1-positive subjects who tested positive for isolated anti-HBc but negative for anti-HBe was low and was comparable to that in subjects who tested negative for anti-HBc. This finding suggests that testing for anti-HBc alone may not be a reliable assessment of protection from HBV infection. HIV-1/HCV coinfection may be associated with impaired responses to hepatitis B vaccine, and evaluation of strategies to improve immunogenicity of the vaccine in such individuals is warranted. PMID- 15809902 TI - Compromised B cell responses to influenza vaccination in HIV-infected individuals. AB - BACKGROUND: Yearly influenza vaccination, although recommended for human immunodeficiency virus (HIV)-infected individuals, has not received thorough evaluation in the era of antiretroviral therapy. We assessed the impact of HIV disease on B cell responses to influenza vaccination. METHODS: Sixty-four HIV infected and 17 HIV-negative individuals received the 2003-2004 trivalent inactivated influenza vaccine. Frequencies of influenza-specific antibody secreting cells (ASCs) were measured by enzyme-linked immunospot (ELISPOT) assay, and antibody responses were measured by hemagglutination-inhibition (HI) assay. Memory responses to influenza were measured by ELISPOT assay after polyclonal activation of B cells in vitro. RESULTS: Prevaccination HI titers were significantly higher in HIV-negative than in HIV-infected individuals. Peak HI titers and influenza-specific ASC frequencies were directly correlated with CD4+ T cell counts in HIV-infected individuals. Influenza-specific memory B cell responses were significantly lower in HIV-infected than in HIV-negative individuals and were directly correlated with CD4+ T cell counts. CONCLUSIONS: HIV infection is associated with a weak antibody response to influenza vaccination that is compounded by a poor memory B cell response. CD4+ T cell count is a critical determinant of responsiveness to influenza vaccination, and the contribution of plasma HIV RNA level is suggestive and warrants further investigation. PMID- 15809903 TI - Dendritic and natural killer cell subsets associated with stable or declining CD4+ cell counts in treated HIV-1-infected children. AB - BACKGROUND: Natural killer (NK) cells and plasmacytoid and myeloid dendritic cells (DCs) are depleted, and their function impaired, in advanced adult human immunodeficiency virus (HIV)-1 infection. Studies in perinatally infected children are lacking. METHODS: Percentages of NK cells and plasmacytoid and myeloid DCs were evaluated by flow cytometry. Forty children with perinatal HIV-1 infection were compared with 11 age-matched, uninfected children. Plasmacytoid and myeloid DC function was evaluated by activation-induced cytokine secretion. RESULTS: Virally suppressed children had normal levels of circulating plasmacytoid and myeloid DCs and total NK cells but had sustained depletion of a mature (CD3-/161+/56+/16+) NK cell subset and decreased interferon- alpha secretion by plasmacytoid DCs. Despite similar viral loads, percentages of myeloid and plasmacytoid DCs and mature NK cells were significantly lower in viremic children with a history of decreasing CD4+ cell percentages, compared with children with stable CD4+ cell counts. CONCLUSIONS: Children achieve partial reconstitution of myeloid and plasmacytoid DCs and NK cells during viral suppression; irrespective of viral load, a clinical history of decreasing CD4+ cell percentage is associated with greater depletion of these subsets. We hypothesize that the evaluation of selected innate-immunity effector cells may serve as a marker of CD4+ cell loss in pediatric HIV-1 infection. PMID- 15809904 TI - Viral dynamics of primary HIV-1 infection in Senegal, West Africa. AB - BACKGROUND: Few studies have addressed primary human immunodeficiency virus (HIV) type 1 infection in sub-Saharan Africa, where the epidemic is of a predominantly heterosexual character and is caused by different subtypes. The present study examines the dynamics of viral replication in subjects infected with various HIV 1 subtypes. METHODS: Seven hundred fifty-two HIV-negative Senegalese women at high risk for infection were monitored every 3 months for acute/early HIV infection; 26 infections were identified (23 HIV-1 and 3 HIV-2), with an HIV-1 incidence rate of 3.23 cases/person-years observation. Multiple viral-load measurements were taken for all seroconverters. RESULTS: The mean+/-standard deviation viral load for all subjects during the early stage of infection was 4.13+/-0.66 log10 copies/mL, with an overall decrease of 0.22 log10 copies/mL after the early stage; the viral set point was reached after 12 months of infection. Most subjects had relatively low viral loads during the early stage of infection. HIV-1 CRF02_AG-infected women had a significantly higher mean viral load during the early stage of infection (mean +/- SD, 4.45+/-0.60 log(10) copies/mL) than did non-HIV-1 CRF02_AG-infected women (mean+/-SD, 3.78+/-0.46 log(10) copies/mL) (P=.008). None of the subjects reported symptoms consistent with primary HIV-1 infection. CONCLUSION: Our findings in Senegalese women differ from what have been described for primary HIV-1 infection. Further investigations of primary infections with non-B subtypes are warranted, to better characterize their differences with primary infections with subtype B. PMID- 15809905 TI - Infection with HIV-1 induces a decrease in mtDNA. AB - Cross-sectional studies have suggested that infection with human immunodeficiency virus (HIV) type 1 could reduce the mitochondrial DNA (mtDNA) content of blood cells. We investigated mtDNA content in peripheral blood mononuclear cells (PBMCs) obtained from 36 antiretroviral therapy-naive documented HIV-1 seroconverters, before and after seroconversion. mtDNA content statistically significantly decreased 1 year after seroconversion and showed a nonsignificant decrease during the subsequent 4 years. These findings confirm that infection with HIV-1 may, itself, reduce mtDNA content, at least within PBMCs. This could have implications for the subsequent development of mitochondrial toxicities associated with the use of nucleoside analogue reverse-transcriptase inhibitors. PMID- 15809906 TI - Detection of airborne severe acute respiratory syndrome (SARS) coronavirus and environmental contamination in SARS outbreak units. AB - Severe acute respiratory syndrome (SARS) is characterized by a risk of nosocomial transmission; however, the risk of airborne transmission of SARS is unknown. During the Toronto outbreaks of SARS, we investigated environmental contamination in SARS units, by employing novel air sampling and conventional surface swabbing. Two polymerase chain reaction (PCR)-positive air samples were obtained from a room occupied by a patient with SARS, indicating the presence of the virus in the air of the room. In addition, several PCR-positive swab samples were recovered from frequently touched surfaces in rooms occupied by patients with SARS (a bed table and a television remote control) and in a nurses' station used by staff (a medication refrigerator door). These data provide the first experimental confirmation of viral aerosol generation by a patient with SARS, indicating the possibility of airborne droplet transmission, which emphasizes the need for adequate respiratory protection, as well as for strict surface hygiene practices. PMID- 15809907 TI - Molecular epidemiology of severe acute respiratory syndrome-associated coronavirus infections in Taiwan. AB - BACKGROUND: In 2003, Taiwan experienced a series of outbreaks of severe acute respiratory syndrome (SARS) and 1 laboratory-contamination accident. Here we describe a new phylogenetic analytical method to study the sources and dissemination paths of SARS-associated coronavirus (SARS-CoV) infections in Taiwan. METHODS: A phylogenetic analytical tool for combining nucleotide sequences from 6 variable regions of a SARS-CoV genome was developed by use of 20 published SARS-CoV sequences; and this method was validated by use of 80 published SARS-CoV sequences. Subsequently, this new tool was applied to provide a better understanding of the entire complement of Taiwanese SARS-CoV isolates, including 20 previously published and 19 identified in this study. The epidemiological data were integrated with the results from the phylogenetic tree and from the nucleotide-signature pattern. RESULTS: The topologies of phylogenetic trees generated by the new and the conventional strategies were similar, with the former having better robustness than the latter, especially in comparison with the maximum-likelihood trees: the new strategy revealed that during 2003 there were 5 waves of epidemic SARS-CoV infection, which belonged to 3 phylogenetic clusters in Taiwan. CONCLUSIONS: The new strategy is more efficient than its conventional counterparts. The outbreaks of SARS in Taiwan originated from multiple sources. PMID- 15809908 TI - A prospective study of sexual transmission of human T lymphotropic virus (HTLV)-I and HTLV-II. AB - BACKGROUND: Cross-sectional studies support sexual transmission of human T lymphotropic virus (HTLV)-I/II; however, prospective incidence data, particularly for HTLV-II, are limited. METHODS: A cohort of 85 HTLV-positive (30 with HTLV-I and 55 with HTLV-II) blood donors and their stable (>or=6 months) heterosexual sex partners were followed biannually over the course of a 10-year period. RESULTS: Four of 85 initially seronegative sex partners of HTLV-I and -II carriers seroconverted, for an incidence rate (IR) of 0.6 transmissions/100 person-years (py) (95% confidence interval [CI], 0.2-1.6). This includes 2 HTLV-I transmissions/219 py (IR, 0.9 transmissions/100 py [95% CI, 0.1-3.3]) and 2 HTLV II transmissions/411 py (IR, 0.5 transmissions/100 py [95% CI, 0.06-1.8]), with no significant difference by HTLV type. There were 2 male-to-female (IR, 1.2 transmissions/100 py [95% CI, 0.1-4.3]) and 2 female-to-male (IR, 0.4 transmissions/100 py [95% CI, 0.05-1.6) transmissions. HTLV-I or -II proviral load was 2 log10 lower in newly infected partners than in index positive partners who transmitted HTLV (P=.007). CONCLUSIONS: The incidence of sexual transmission of HTLV-II may be similar to that of HTLV-I, and female-to-male transmission may play a more important role than previously thought. HTLV-I and -II proviral load may be lower in sexually acquired infection, because of a small infectious dose. PMID- 15809909 TI - Effect of heptitis C virus core protein on cellular gene expression: specific inhibition of cyclooxygenase 2. AB - Hepatitis C virus (HCV) core protein plays a significant role in the alteration of cellular gene expression. We expressed HCV core protein using a tetracycline inducible expression system in HeLa cell lines. Profiles of gene expression in cells expressing the HCV core protein were compared with those in control cells by use of microarray analysis. Cells expressing the HCV core protein showed 86 down-regulated and 41 up-regulated genes, compared with control cells. One gene affected was cyclooxygenase 2 (COX-2). Levels of both COX-2 RNA and the Cox-2 protein were significantly inhibited after the expression of HCV core protein in HeLa cells. Similar results were obtained in hepatoma cells and in a functional assay that measured the production of the Cox-2 protein in response to a mitogenic stimulus. The inhibition of the Cox-2 protein could serve as a means of muting the cellular inflammatory response during HCV infection. Correlation of these findings with analysis of clinical specimens from chronically infected patients should lend further significance to the down-regulation of the inflammatory response via Cox-2. PMID- 15809910 TI - Antidiarrheal effects of L-histidine-supplemented rice-based oral rehydration solution in the treatment of male adults with severe cholera in Bangladesh: a double-blind, randomized trial. AB - BACKGROUND: Because of the antisecretory potential of L-histidine in the intestinal tract, its antidiarrheal effects were determined in cholera. METHODS: In a double-blind trial of 126 adult male patients with cholera, L-histidine (2.5 g/L) was mixed with a rice-based oral rehydration solution (ORS) and administered to 62 patients; 64 patients received the same ORS without L-histidine. All patients received ciprofloxacin at a dosage of 500 mg every 12 h for 72 h. Fluid output (of stool, urine, and vomit) and intake (of ORS, water, and intravenous fluid) were determined every 8 h for 72 h. RESULTS: Administration of ORS with L histidine significantly (P<.05) reduced the frequency of stool output during 32 64 h after initiation of ORS treatment, compared with that in patients given ORS without L-histidine ([all data are means+/-SD] 32-48 h, 11.5+/-6.9 mL/kg vs. 18.8+/-16.0 mL/kg; 40-48 h, 6.7+/-4.4 mL/kg vs. 11.5+/-9.7 mL/kg; and 56-64 h, 6.3+/-5.8 mL/kg vs. 7.8+/-4.1 mL/kg). An overall reduction of 22% in the volume of stool was observed in patients given ORS without L-histidine. The amount of required unscheduled intravenous fluid was lower in patients given ORS with L histidine, compared with that in patients given ORS without L-histidine (0-24 h, 82.5+/-44.4 mL/kg vs. 158.6+/-72.2 mL/kg [P<.01]; and 24-48 h, 41.6+/-40.4 mL/kg vs. 52.5+/-22.1 mL/kg [P>.05]). Administration of ORS with L-histidine also significantly reduced (P<.05) the intake of ORS and the duration of illness. No adverse effects were observed in these patients. CONCLUSIONS: L-histidine reduces the weight of stool and the frequency of stool output in cholera and could be a useful and safe adjunct treatment that will increase the success rate of ORS and antibiotic therapy in cholera. PMID- 15809911 TI - Neutralization of endotoxin by a phospholipid emulsion in healthy volunteers. AB - BACKGROUND: An approach to endotoxin (lipopolysaccharide [LPS]) blockade makes use of the ability of lipoproteins, via surface phospholipids, to bind and neutralize LPS. The aim of the present study was to determine whether the intravenous administration of a protein-free, phospholipid-rich emulsion is an effective method for neutralizing the effects of LPS in healthy persons. METHODS: This was a double-blind, placebo-controlled study in 20 volunteers. Volunteers received Escherichia coli endotoxin (2 ng/kg) intravenously 2 h into a 6-h infusion of either emulsion (210 mg/kg) or placebo (Intralipid diluted 1 : 64). RESULTS: The volunteers who received emulsion had a lower mean clinical score (P<.01), temperature (P<.05), pulse rate (P<.05), neutrophil count (P<.05), tumor necrosis factor- alpha level (P<.05), and interleukin-6 level (P<.05) than did the volunteers who received placebo. Response was related to serum phospholipid level. The greatest effects were observed in the volunteers achieving phospholipid levels of approximately 500 mg/dL or higher. CONCLUSION: Phospholipid emulsion attenuates the clinical and laboratory effects associated with the administration of LPS in humans, suggesting a novel approach to the treatment of endotoxemia. PMID- 15809912 TI - Burden of community-onset Escherichia coli bacteremia in seniors. AB - BACKGROUND: Although Escherichia coli is a well-recognized cause of urinary tract infection in seniors, little is known about the burden of invasive E. coli infection in this population. METHODS: We conducted a population-based cohort study of 46,238 noninstitutionalized Group Health Cooperative members>or=65 years of age to ascertain incidences of community-onset E. coli bacteremia and, for comparison, pneumococcal bacteremia, and we then performed a case-control study to identify risk factors for community-onset E. coli bacteremia. RESULTS: The overall rate of community-onset E. coli bacteremia in the study cohort was 150 cases/100,000 person-years, which was approximately 3 times higher than the rate of pneumococcal bacteremia. In the case-control study, urinary catheterization and urinary incontinence were the only factors associated with an increased risk of E. coli bacteremia in men (62 cases), whereas cancer, renal failure, congestive heart failure, coronary artery disease, and urinary incontinence were associated with an increased risk of E. coli bacteremia in women (119 cases). CONCLUSIONS: E. coli appears to be the leading cause of community-onset bacteremia in seniors, and, on the basis of these rates, we estimate that 53,476 cases occur in noninstitutionalized seniors each year in the United States. Community-onset E. coli bacteremia in seniors is, therefore, an infection of public health importance. PMID- 15809913 TI - An outbreak of Pontiac fever with respiratory distress among workers performing high-pressure cleaning at a sugar-beet processing plant. AB - BACKGROUND: In August 2000, the Minnesota Department of Health was notified of and investigated an outbreak of febrile respiratory illness among workers at a sugar-beet processing plant. METHODS: A case was defined as fever and respiratory symptoms occurring in a worker at the sugar-beet plant on or after 31 July 2000. Case patients were interviewed, medical and work records were reviewed, and clinical samples were obtained. The plant was inspected, and environmental samples were collected. RESULTS: Fourteen of 15 case patients performed high pressure water cleaning in the confined space of an evaporator vessel. Symptoms included fever and chills (100%), chest tightness (93%), cough (80%), and shortness of breath (73%). In case patients, median temperature was 39.4 degrees C, median oxygen saturation was 93%, and median white blood cell count was 12x10(3) cells/ mu L. Four (29%) of 14 case patients showed evidence of Legionella pneumophila exposure, according to serologic testing. Water sources contained up to 10(5) cfu/mL of L. pneumophila and 22,200 endotoxin units/mL. CONCLUSIONS: Outbreak features were consistent with Pontiac fever. Respiratory symptoms, which are atypical for Pontiac fever, could be attributed to a high exposure dose of L. pneumophila from confined-space aerosolization or to endotoxin exposure. This outbreak demonstrates the potential occupational hazards for those performing high-pressure cleaning in confined spaces. PMID- 15809914 TI - The role played by tumor necrosis factor during localized and systemic infection with Streptococcus pneumoniae. AB - Tumor necrosis factor (TNF) has been proposed as a major mediator of host resistance in murine models of Streptococcus pneumoniae infection; in humans, anti-TNF therapies have been implicated in increased susceptibility to pneumococcal infection. Here, we use nonlethal (serotype 6B) and lethal (serotype 3) S. pneumoniae, neutralizing monoclonal antibodies to TNF, and TNF gene deficient mice to reexamine the role played by TNF in antistreptococcal responses. After nonlethal challenge, primary resistance and all examined parameters of the cellular inflammatory response occurred independently of TNF activity. After lethal challenge, TNF deficiency resulted in more-rapid death but did not affect lung inflammation. However, the livers of the TNF gene-deficient mice, but not of the control mice, exhibited extensive signs of systemic disease. TNF, therefore, is dispensable for a complete cellular pulmonary inflammatory response to S. pneumoniae infection but enhances survival from disseminated lethal infection, at least in part by delaying systemic organ damage. PMID- 15809915 TI - Impaired systemic production of prostaglandin E2 in children with cerebral malaria. AB - Prostaglandins (PGs) are important mediators of macrophage activity, vascular permeability, fever, erythropoiesis, and proinflammatory responses to infection. Our recent studies have shown that plasma levels of bicyclo-PGE2 (a stable end product of PGE2 metabolism) and leukocyte cyclooxygenase (COX)-2 gene expression are suppressed in children with malarial anemia. Since the role of PGs as immunomodulators of human cerebral malaria (CM) has not been examined, we investigated urinary levels of bicyclo-PGE2/creatinine in children with varying clinical outcomes of CM. Among parasitemic children, those with asymptomatic parasitemia had the highest levels of bicyclo-PGE2/creatinine, whereas those with CM had significantly lower levels of bicyclo-PGE2. Systemic levels of bicyclo PGE2/creatinine were not significantly associated with parasitemia, hemoglobin levels, or levels of the PG-regulatory cytokine tumor necrosis factor- alpha but were positively correlated with levels of interleukin-10. The results presented here show that impaired systemic production of PGE2 is associated with adverse outcomes of CM, whereas elevated levels of PGE2 in asymptomatic parasitemia suggest a potential role for PGs in protective immunity. PMID- 15809916 TI - Fragmented population structure of plasmodium falciparum in a region of declining endemicity. AB - BACKGROUND: The population genetic structure of Plasmodium falciparum differs between endemic regions, but the characteristics of a population recently fragmented by effective malaria control have been unknown. METHODS: Genotypic analysis of 10 microsatellite loci widely separated in the parasite genome was conducted on 288 P. falciparum isolates from 8 foci in Malaysian Borneo, a region in which malaria incidence has been progressively reduced. RESULTS: Within all P. falciparum foci, moderate levels of allelic diversity were found, but levels of multilocus linkage disequilibrium were extremely variable. The population with the highest proportion of mixed-clone infections also had the highest allelic diversity and nonsignificant linkage disequilibrium. In contrast, several populations showed evidence of clonal expansion, and one offshore island population had exceptionally high levels of linkage disequilibrium. Genetic differentiation between many populations was very high and strongly associated with the geographical distance between them. CONCLUSIONS: High levels of differentiation and contrasting population structure among P. falciparum populations in Malaysian Borneo indicate that they are genetically independent. This supports the feasibility of individually eradicating the remaining P. falciparum foci. PMID- 15809917 TI - Ciprofloxacin resistance does not affect duration of domestically acquired campylobacteriosis. PMID- 15809919 TI - Safety of stavudine during pregnancy. PMID- 15809920 TI - Hepatitis C virus genotype and the natural history of HIV-1 disease: potential role of GB virus C in the hemophiliac population. PMID- 15809923 TI - Amphotericin B lipid complex in the treatment of invasive fungal infections: results of the Collaborative Exchange of Antifungal Research (CLEAR), an industry supported patient registry. PMID- 15809924 TI - Treatment of Candida infections with amphotericin B lipid complex. AB - The efficacy and renal safety of amphotericin B lipid complex (ABLC) were assessed in >900 patients with candidiasis. Overall, a favorable clinical response (cured or improved) was observed in 61% of patients infected with Candida species only, in 62% of patients infected with C. albicans, and in 61% of patients infected with a non-albicans Candida species. Clinical responses were similar in patients infected with invasive C. albicans and non-albicans Candida species (63% and 62%, respectively). Similarly, response rates of 60% and 59% were observed in patients infected with noninvasive C. albicans and non-albicans Candida species, respectively. Compared with patients who received lower doses of ABLC, patients who required higher doses of ABLC because of more-virulent infections did not demonstrate significant renal impairment, as assessed by end of-therapy changes in serum creatinine level from baseline (median, 0.1 mg/dL; range, -3.9 to 2.4 mg/dL), incidence of serum creatinine doubling (16%), and need for new dialysis (7%). These data indicate the safety and efficacy of ABLC in treating candidiasis. PMID- 15809925 TI - Amphotericin B lipid complex in the management of invasive aspergillosis in immunocompromised patients. AB - The efficacy and renal safety of amphotericin B lipid complex (ABLC) were assessed in 398 patients with invasive aspergillosis. The most common underlying conditions were hematopoietic stem-cell transplantation (101/398 [25%]), hematologic malignancy (101/398 [25%]), and solid-organ transplantation (109/398 [27%]). The most common reason for administration of ABLC was lack of response to prior antifungal therapy. Overall, 65% of patients had a favorable clinical response: 44% were cured or improved, and 21% were stabilized. Clinical responses were similar for patients who received ABLC as either first-line or second-line therapy. One hundred forty-four patients whose results could be evaluated received ABLC concurrently with or after therapy with itraconazole. No antagonism was observed when therapy with ABLC followed therapy with itraconazole. Patients infected with Aspergillus terreus, an innately polyene-resistant species, experienced a 37% response rate. Changes in serum creatinine levels were not clinically significant in most patients; however, dialysis was initiated in 7 patients, of whom 6 had prior antifungal therapy or preexisting renal disease. Analysis of this large database demonstrated the efficacy and safety of ABLC in the treatment of invasive aspergillosis. PMID- 15809926 TI - Treatment of non-Aspergillus moulds in immunocompromised patients, with amphotericin B lipid complex. AB - An assessment was made of the efficacy and renal safety of amphotericin B lipid complex (ABLC) in the treatment of patients with invasive fungal infections caused by moulds other than Aspergillus species, on the basis of a retrospective analysis of data from the Collaborative Exchange of Antifungal Research (CLEAR) database. Data from CLEAR for 64 patients with zygomycosis were published previously. The database was further queried and yielded results for 28 patients with fusariosis and 84 patients infected with other non-Aspergillus moulds. Of 26 patients with fusariosis whose results could be evaluated, 46% (n = 12) were cured or improved, and an additional 12% (n = 3) were stable. Of 79 patients infected with other non-Aspergillus moulds whose results could be evaluated, 61% (n = 48) were cured or improved, and an additional 15% (n = 12) were stable. In an area with little guidance for therapy, the CLEAR data indicate that ABLC can be an effective broad-spectrum treatment choice for several invasive and refractory non-Aspergillus mould infections. PMID- 15809927 TI - Successful use of amphotericin B lipid complex in the treatment of cryptococcosis. AB - The efficacy and renal safety of amphotericin B lipid complex (ABLC) injection were assessed in 106 patients with cryptococcal infection. Eighty-three patients (78%) had a central nervous system (CNS) infection. Of these patients, 20 initiated azole therapy concomitantly with ABLC therapy, and 7 had received prior azole therapy, which continued during administration of ABLC. Clinical response (cured or improved) was achieved in 67 (66%) of 101 patients whose results could be evaluated. Response rates were 65% (51/78) for patients with a CNS infection and 70% (16/23) for patients without a CNS infection. The response rate for patients with HIV infection was 58% (30/52). Response rates were 56% (19/34) for patients who were refractory to prior antifungal therapy, 65% (11/17) for patients who were intolerant of prior antifungal therapy, 60% (3/5) for patients with underlying renal disease who received prior antifungal therapy, 76% (25/33) for patients with underlying renal disease who did not receive prior antifungal therapy, and 73% (8/11) for patients with no renal disease who did not receive prior antifungal therapy. A mean serum creatinine level decrease of 0.02 mg/dL occurred. ABLC was an effective treatment for cryptococcal infection in immunocompromised patients. PMID- 15809928 TI - Study of renal safety in amphotericin B lipid complex-treated patients. AB - To investigate the renal safety of amphotericin B lipid complex (ABLC), records from 3514 ABLC-treated patients with fungal infections were reviewed. The median change in predicted creatinine clearance (CCr) from baseline to the end of therapy was -3 mL/min (range, -119 to 118 mL/min); doubling of serum creatinine (S-Cr) level occurred in 13% of patients, and new dialysis was needed for 3% of patients. Patients with underlying renal disease who had received prior antifungal therapy demonstrated a median CCr of 0.5 mL/min (range, -107 to 52 mL/min). Despite increased risk for renal impairment in allogeneic hematopoietic stem-cell transplant recipients, only 17% of patients demonstrated end-of-therapy doubling of S-Cr levels, and the median change in CCr was -10 mL/min (range, -107 to 108 mL/min). In ABLC-treated patients, concomitant treatment with potentially nephrotoxic agents and a baseline S-Cr level of <2 mg/dL were factors predisposing for the development of nephrotoxicity. These data provide evidence that ABLC may be used safely to treat patients who are at increased risk for renal impairment. PMID- 15809931 TI - Is partial breast irradiation a step forward or backward? AB - Partial-breast irradiation (PBI) may prove to be a significant advance in the treatment of selected patients with early-stage breast cancer. However, because PBI treats a much smaller volume of breast tissue than does conventional whole breast irradiation and the biologic equivalent dose delivered to the tumor may be considerably lower, it is possible that this treatment approach may increase the risk of recurrence. There may also be an increased risk of toxicity using PBI techniques that have significant dose inhomogeneity. Despite these uncertainties, many patients in the United States are currently treated with PBI outside of a clinical study. Showing that PBI is safe and effective requires having results from large clinical trials. If such trials are supported and completed, the benefits and limitations of PBI are likely to be elucidated, and this treatment technique, properly refined, may well prove to be an advance in breast cancer care. If instead PBI is adopted ad hoc within the radiation oncology community without proper testing, it is likely that poor outcomes will be underreported and many patients will receive suboptimal therapy. This would be a big step backward in the role of radiation therapy in breast cancer treatment. PMID- 15809929 TI - A novel linkage to generalized vitiligo on 4q13-q21 identified in a genomewide linkage analysis of Chinese families. AB - Generalized vitiligo is a common, autoimmune, familial-clustering depigmentary disorder of the skin and hair that results from selective destruction of melanocytes. Generalized vitiligo is likely a heterogeneous disease, with five susceptibility loci reported so far--on chromosomes 1p31, 6p21, 7q, 8p, and 17p13 -in white populations. To investigate vitiligo susceptibility loci in the Chinese population, we performed a genomewide linkage analysis in 57 multiplex Chinese families, each with at least two affected siblings, and we identified interesting linkage evidence on 1p36, 4q13-q21, 6p21-p22, 6q24-q25, 14q12-q13, and 22q12. Subsequently, to extract more linkage information, we investigated our initial genomewide linkage findings in a follow-up analysis of 49 new families and additional markers. Our initial genomewide linkage analysis and our subsequent follow-up analysis have identified a novel linkage to vitiligo on 4q13-q21, with highly significant linkage evidence (a nonparametic LOD score of 4.62 [P=.000003] and a heterogeneity LOD score of 4.01, under a recessive inheritance model), suggesting that 4q13-q21 likely harbors a major susceptibility locus for vitiligo in the Chinese population. We observed a minimal overlap between the linkage results of our current genomewide analysis in the Chinese population and the results of previous analyses in white populations, and we thus hypothesize that, as a polygenic disorder, vitiligo may be associated with great genetic heterogeneity and a substantial difference in its genetic basis between ethnic populations. PMID- 15809932 TI - Intraoperative electrons. AB - Intraoperative radiotherapy (IORT) has been used for many years for treating patients with various locally-advanced malignancies, usually combined with external-beam radiation therapy (EBRT). Long-term results confirm that IORT improves local control, which is generally associated with increased survival. Recently, electron-beam IORT has been used as the sole treatment for patients with earlier-stage cancers, especially for breast tumors, with extremely promising results. Most of this work has been done at the European Institute of Oncology in Milan. We report the rationale and techniques of the use of electron intraoperative treatment (ELIOT) and the results of our different clinical studies. In our opinion, ELIOT may be an excellent alternative to EBRT for the treatment of patients with early-stage breast cancer. However, intensive long term follow-up is needed to fully evaluate local control and possible side effects. PMID- 15809933 TI - TARGeted Intraoperative radiotherapy (TARGIT): an innovative approach to partial breast irradiation. AB - A revolution is challenging the dogma that local treatment for all patients with breast cancer treated with breast conservation therapy must include postoperative radiotherapy delivered to the whole breast. Such prolonged postoperative radiotherapy is a burden to patients and hospitals and forces many women to chose mastectomy instead. Furthermore, for patients receiving chemotherapy, the start of conventional radiotherapy may be delayed so long as to increase the risk of local relapse. These problems might be eliminated if effective radiotherapy could be given as a single treatment intraoperatively, immediately after the surgery. Local recurrence after breast-conserving surgery usually occurs in the portion of the breast in the immediate proximity of the tumor, even when radiotherapy is omitted. Therefore, it should usually be possible to restrict radiotherapy to only the area adjacent to the tumor in selected women. Based on this premise, we have devised a new technique of partial breast irradiation, with the intention of completing all local treatment in a single session. In this article, we elaborate on the rationale and on the different methods of delivering intraoperative radiotherapy. If this approach is validated in ongoing randomized trials, it could save time, money, and breasts. PMID- 15809934 TI - External-beam partial-breast irradiation. AB - Although most studies treating patients with partial-breast irradiation have used brachytherapy, giving such treatment with external-beam techniques has many potential advantages. However, there is only limited published experience using this approach. These include a randomized trial of partial-breast and whole breast irradiation performed at the Christie Hospital in Manchester, England, and pilot studies (using much more rigorous selection criteria and sophisticated treatment planning) from groups at the University of Southern California, New York University (using prone positioning of patients), and the William Beaumont Hospital (using the supine position). A multi-institutional pilot trial based on the latter technique has been completed, which was designed to test the feasibility of using this approach in the cooperative oncology group setting. The unprecedented rapidity with which the study completed its target accrual indicates the degree of interest in this approach. This review focuses on the rationale and the reported studies of external-beam partial-breast radiation and identifies some specific issues and remaining problems associated with this approach. PMID- 15809935 TI - The MammoSite balloon brachytherapy catheter for accelerated partial breast irradiation. AB - Whole-breast irradiation after lumpectomy for early-stage breast cancer appears to prevent relapse only at the site of initial involvement, and therefore it is likely that radiation can safely be limited to tissues immediately surrounding the excision cavity. Accelerated partial-breast irradiation (APBI) with the MammoSite catheter (Proxima Therapeutics, Alpharetta, GA) has been proposed as a means through which APBI can be offered to a wide range of patients. Although sufficient follow-up is not yet available regarding local control, survival, late toxicity, and long-term cosmetic results, early results are promising. This article reviews the current literature reflecting treatment, dose optimization, outcomes, and toxicity of APBI with the MammoSite brachytherapy catheter. PMID- 15809936 TI - Breast brachytherapy: North American experience. AB - Accelerated partial breast irradiation (APBI) has been investigated for over a decade as a potential alternative adjuvant treatment approach after lumpectomy for women with early-stage breast cancer. The rationale for APBI is based on pathologic data regarding the spread of cancer within the breast and study of the patterns of in-breast recurrence after breast-conserving therapy performed with or without whole-breast irradiation. This report reviews the North American experience using interstitial brachytherapy for APBI. Studies achieving low failure rates have universally been distinguished from those with high failure rates by requiring documented microscopically negative surgical margins, using a target definition consisting of the lumpectomy cavity plus a 1- to 2-cm margin, and having a rigorous quality assurance program to assure target coverage. We conclude that APBI brachytherapy programs that include all of these components have great potential to overcome many of the barriers that have prevented women from pursuing standard breast-conserving therapy. PMID- 15809937 TI - Brachytherapy for partial breast irradiation: the European experience. AB - There has been great interest in Europe in treating selected patients with early stage breast cancer with accelerated partial breast irradiation (APBI) using interstitial brachytherapy. We review the results of these studies. Five early studies had local recurrence rates of 6% to 37%, reflecting suboptimal patient selection, target definition, and quality assurance procedures. Five more recent studies performed using much more stringent approaches have had recurrence rates of 0% to 7% with 84% to 92% of patients having excellent or good cosmetic results. The 3-year results of a single-institution phase III trial suggest that partial-breast brachytherapy performed in this manner yields local control that is very similar to that of whole-breast irradiation. Recently, a phase III multicenter protocol comparing these 2 modalities has been activated in Europe by the Breast Cancer Working Group of the Groupe Europeen de Curietherapie-European Society for Therapeutic Radiology and Oncology. As data from this and other trials mature, they will address and refine issues of patient selection, target volume definition, total dose, and fractionation and hopefully support the implementation of APBI into routine clinical practice. PMID- 15809938 TI - Lessons of studies of breast-conserving therapy with and without whole-breast irradiation for patient selection for partial-breast irradiation. AB - Partial-breast irradiation is a promising approach for treating patients with early-stage invasive breast cancer. There are still few studies of partial-breast irradiation that address critical issues, such as criteria for patient selection. However, an enormous body of experience has been accumulated regarding the results in patients treated with "conventional" breast-conserving therapy using whole-breast irradiation and for patients treated with conservative surgery without radiotherapy of any kind. This article will therefore review portions of this existing experience that may aid the development of partial-breast irradiation and give perspective on the overall role that partial-breast irradiation may ultimately play in our therapeutic armamentarium. PMID- 15809941 TI - Prophylaxis for thromboembolic disease: recommendations from the American College of Chest Physicians--are they appropriate for orthopaedic surgery? PMID- 15809942 TI - Primary exeter total hip arthroplasty in patients with small femurs: a minimal of 10 years follow-up. AB - One hundred and twelve hips in 101 Chinese patients received primary cemented Exeter total hip arthroplasty (Howmedica, Hong Kong). The implant had a collarless, double-tapered and polished stem. Seventy-five hips in 67 patients were available for assessment. The average age at operation was 63.7 years. The commonest diagnosis was avascular necrosis. The average follow-up was 12.8 years (range, 10 to 16.5 years). The Harris score improved from a preoperative average of 39.8 to 82.3 at the last follow-up. Stem subsidence within the cement mantle occurred in 9.3% of the cases and none of these stems had other evidence of loosening. Eight hips (six stems and two cups) in seven patients were revised because of loosening at five to 14 years postoperatively. The survival rate was 93.3% and 86% at 10 and 15 years respectively. Oversized stems and incomplete cement mantle might account for the loosening. Stems of a smaller size have been made available to this population and hopefully this will improve the results further. PMID- 15809944 TI - Arthroplasty in the octogenarian: quantifying the risks. AB - The objective of this study was to determine the risk of complications after primary total hip and knee arthroplasties in octogenarians. Using administrative data, we compared the rate of complications for octogenarians vs patients aged 65 to 79 years who underwent total hip or knee arthroplasty in Ontario between 1993 and 1997. Octogenarians were 3.4 times more likely to die, 2.7 times more likely to sustain a myocardial infarction, and 3.5 times more likely to develop pneumonia. Postoperative confusion and urinary tract infections were also significantly more common in octogenarians. Octogenarians are at higher risk for death and serious complications after primary joint arthroplasty. However, the overall event rate remains low and total joint arthroplasty surgery should continue to be offered to these individuals provided that the complication rates are acceptable to the patients and their families. PMID- 15809943 TI - Leg axis after computer-navigated total knee arthroplasty: a prospective randomized trial comparing computer-navigated and manual implantation. AB - To compare the alignment after computer-navigated total knee arthroplasty, 52 patients were randomly allocated to 2 groups. Twenty-seven patients received a total knee arthroplasty with the aid of a kinematic computer-navigation system, and 25 patients received a total knee arthroplasty with the conventional method. Both groups were well balanced concerning demographic data and preoperative scores. At 3-month follow-up, the mechanical alignment of the leg reached the desired straight axis in more cases with the computer-navigated implantation. This difference was statistically significant. The femoral and tibial mechanical anteroposterior axis and the femoral and tibial sagittal tilt (slope) measured on sagittal x-rays were not significantly improved in this patient group. PMID- 15809945 TI - Extended trochanteric osteotomy for 2-stage revision of infected total hip arthroplasty. AB - We evaluate the rate of osteotomy healing, implant stability, and eradication of infection when an extended trochanteric osteotomy, with interval placement of an antibiotic-impregnated cement spacer and delayed osteotomy fixation, is used to treat the chronically infected total hip arthroplasty. Thirteen cases were followed for a minimum of 2 years. All patients had complete healing of the extended trochanteric osteotomy within 6 months. At an average follow-up of 39 months, recurrent infection occurred in 3 (23%) patients. Femoral component subsidence of 5 mm occurred in 2 patients, both of which had recurrent infection. Extended trochanteric osteotomy with interval placement of an articulating antibiotic-impregnated cement spacer and delayed osteotomy fixation permits reliable healing of the osteotomy. PMID- 15809946 TI - Revision total knee arthroplasty with modular components inserted with metaphyseal cement and stems without cement. AB - The clinical and radiographic outcomes of 50 consecutive revision total knee arthroplasties in 47 patients, placed with metaphyseal cemented femoral and tibial components with press-fit cementless stems, were reviewed at 36-month average follow-up. Revision was performed for aseptic loosening (11/50), infection (17/50), periprosthetic fracture (8/50), component failure (6/50), instability (6/50), and malalignment (2/50). The press-fit cementless stems were 80 to 160 mm in length and tightly contacted the endosteum of the metadiaphyseal areas. Four (9%) knees were re-revised for infection, zero for aseptic loosening. The average modified Hospital for Special Surgery knee score improved from 49 to 87. One patient (2%) reported thigh pain, and 1 reported leg pain. Metaphyseal cemented revision total knee components with press-fit cementless femoral and tibial stems were not associated with significant thigh and leg pain. PMID- 15809947 TI - Precoated femoral component with proximal and distal centralizers: results at 5 to 12 years. AB - One study, confounded by the use of crystalline polyethylene and ceramic heads, reported a high rate of early failure of a precoated femoral component with proximal and distal centralizers. The present study reports the prospective clinical and radiographic results, and 10-year survival data of 166 consecutive hybrid total hip arthroplasties using this femoral component. An A or B cement grade was obtained in 93% of hips. At a mean follow-up time of 7 years (range, 5 12 years), there were only 4 (2.4%) femoral failures. Ten-year survival of this component was 95% (confidence interval, 94%-99%). The rate of radiographic failure and revision of this component implanted with conventional polyethylene and cobalt chrome heads is similar to that reported with other "modern" femoral components. PMID- 15809948 TI - Cryo/Cuff compared to epidural anesthesia after knee unicompartmental arthroplasty: a prospective, randomized and controlled study of 60 patients with a 6-week follow-up. AB - The aim of this prospective, randomized, and controlled study was to evaluate the efficacy of cold compressive dressings (Cryo/Cuff) and epidural anesthesia (EDA) in the postoperative management of primary unicondylar knee arthroplasty. Sixty patients (61 knees) were randomized into 3 groups. No significant difference between groups was detected with respect to subjective pain, bleeding, swelling, range of motion, and function. The consumption of morphine was, however, significantly higher in the control group the first 24 hours than both the EDA group (P < .001) and the Cryo group (P = .028). There was no significant difference in morphine consumption between the 2 treatment groups. Based on the results of this study, Cryo/Cuff seems to be a rational, effective, risk-free, and well-tolerated alternative to EDA to reduce pain and morphine after unicondylar knee arthroplasty. PMID- 15809949 TI - Postoperative management after total hip and knee arthroplasty. AB - Despite major advances in the field of total joint arthroplasty, a standardized postoperative management protocol currently does not exist following total hip arthroplasty (THA) and total knee arthroplasty (TKA). A survey was mailed to the active members of the American Association of Hip and Knee Surgeons to investigate issues such as postoperative rehabilitation and activity restriction. The information derived from this survey provides the total joint surgeon with a compilation and consensus of responses that can serve as the foundation for a standardized postoperative protocol for THA and TKA surgery. PMID- 15809950 TI - Effect of tourniquet pressure on perioperative blood loss associated with cementless total knee arthroplasty: a prospective, randomized study. AB - Perioperative blood loss is an issue of concern in cementless total knee arthroplasty (TKA). We randomly assigned 57 patients (60 knees) who underwent cementless TKA treatment to 2 groups with tourniquet pressure (TP) of either 350 mm Hg (30 knees) or systolic blood pressure (SBP) plus 100 mm Hg (30 knees) to assess the effect of TP on blood loss. The mean TP in the SBP plus 100 mm Hg group was 238 +/- 13 mm Hg. There was no significant difference between groups. Red blood cell count, hemoglobin levels, and hematocrit recovered to preoperative levels by 3 months after surgery in both groups. In conclusion, we recommend using a TP of 100 mm Hg above SBP during TKA, rather than using the conventional TP of 350 mm Hg. PMID- 15809951 TI - Failure of free vascularized fibular graft for osteonecrosis of the femoral head: a histopathologic study of 6 cases. AB - The rationale for the use of free vascularized fibular graft for osteonecrosis of the hip is based on the assumption that the graft will provide support and blood supply to the osteonecrotic head. We report the histopathologic features of 6 consecutive osteonecrotic heads with failed vascularized fibular grafts. Three grafts were partially viable, and 3 were necrotic. The medullary canal was completely necrotic in 4 grafts. A thin layer of new, viable periosteal bone and bone remodeling was present at the surface of 4 grafts. New bone applied to the surface of the graft, especially in the 3 necrotic ones, suggests that the new bone originated from the host bed and not from the graft. Therefore, the risks and benefits of the vascular anastomosis, which in these cases provided no viability to the graft, should be re-evaluated. These findings were observed in failed cases and cannot be extrapolated to successful ones. PMID- 15809952 TI - Cemented hip arthroplasty with a novel cerclage cable technique for unstable intertrochanteric hip fractures. AB - Thirty-nine consecutive patients with unstable three and four part intertrochanteric hip fractures were treated with cemented bipolar hip arthroplasty. A standard length primary femoral component was used with a novel technique of cerclage fixation of the trochanteric bone fragments allowing retention of the femoral calcar. At one year minimum follow up, there was no loosening or subsidence of the femoral components. All trochanters healed. One dislocation and one deep infection occurred. Unstable three and four part hip fractures can be treated with a standard femoral stem and cerclage cabling of the trochanters. The technique allows safe early weight bearing on the injured hip and had a relatively low rate of complications in our series. PMID- 15809953 TI - Total knee arthroplasty for salvage of failed internal fixation or nonunion of the distal femur. AB - Seventeen patients with a mean age of 66 years had total knee arthroplasty (TKA) performed to salvage failed internal fixation or nonunion of the distal femur. Two patients died at less than 2 years, both of whom were unrevised. The remaining 15 were followed for a mean of 5 years. Three arthroplasties failed, 1 by infection and 2 by mechanical failure. The 5-year survivorship free of revision for aseptic failure was 91% (72%-100%). There were 5 intraoperative complications (29%) and 5 postoperative complications (29%). The mean Knee Society pain score improved from 2 (range, 0-10) to 89 (range, 63-97), and the mean functional score improved from 2 (range, 0-25) to 45 (range, 10-90). TKA provided reliable pain relief and functional improvement for the great majority of patients. Functional scores and prosthesis survivorship, however, were inferior to those reported for primary TKA. The surgeries were difficult, and intraoperative and postoperative complications were common. PMID- 15809954 TI - The long-term wear of retrieved McKee-Farrar metal-on-metal total hip prostheses. AB - Twenty-four cobalt-chrome alloy McKee-Farrar matching acetabular and femoral components were retrieved at revision total hip arthroplasty. The average time in situ was 16 years. Wear and loss of sphericity was very low. Polishing wear (type 1), fine abrasive (type 2), multidirectional dull abrasive (type 3), and unidirectional dull abrasive wear (type 4) of the articulating surfaces were identified. The mean percent area of femoral heads occupied by types 2, 3, and 4 wear was 18%, 5%, and 2%, respectively. There was no association between the type and distribution of wear and the time in situ. Impingement damage was evident on 9 implant pairs. The extent and types of wear described in this paper will be useful when analyzing the patterns of surface damage of newer designs of metal-on metal articulations. PMID- 15809955 TI - The nonoperative management of periprosthetic fractures associated with the Birmingham hip resurfacing procedure. AB - We reviewed the outcome of 7 patients sustaining undisplaced femoral neck periprosthetic fractures associated with the Birmingham hip resurfacing procedure. All fractures presented within 4 months of initial surgery. All fractures were treated conservatively with a period of nonweight bearing. The 7 patients were assessed clinically and radiologically at a minimum of 16 months postfracture. All fractures had fully united. There was no evidence of impaired function or abnormal radiological findings after this conservative treatment strategy in 6 patients. One patient had marked femoral neck narrowing but no functional impairment. PMID- 15809956 TI - Ten to twelve-year results with the Zweymuller cementless total hip prosthesis. AB - Between January 1987 and December 1990, 221 Zweymuller cementless total hip arthroplasties were performed in 211 patients with idiopathic osteoarthritis. A total of 136 patients (142 prostheses) were evaluated at a mean follow-up of 134 months (SD 9.5). The study group consisted of 78 Hochgezogen and 64 Stepless stem prostheses, all with a threaded titanium cup and ceramic head. No clinical and radiological differences were found between the 2 stem prostheses. Seven cups had been revised because of aseptic loosening; 17 cups showed radiolucent lines, osteolysis, or migration. Mean linear polyethylene wear of 105 (74%) cups was 0.46 mm (SD 0.27), with an annual wear of 0.04 mm (SD 0.02). Wear did not correlate with pain, cup migration, radiolucent lines, or osteolysis. Cumulative survival was 96%. Zweymuller cementless total hip arthroplasty showed good midterm results. PMID- 15809957 TI - Radiographic evaluation of a monoblock acetabular component: a multicenter study with 2- to 5-year results. AB - Serial radiographs of a porous tantalum monoblock acetabular cup design were evaluated for cup stability and signs of successful osteointegration. Of 574 primary consecutive total hip replacements in 542 patients performed by 9 surgeons at 7 hospitals, 414 cases were available for minimum 2-year follow-up. Follow-up averaged 33 months and ranged from 24 to 58 months. Postoperative radiographs revealed acetabular gaps in 100 zones in 80 (19%) hips: 29 in zone I, 67 in zone II, and 4 in zone III. At last follow-up, 84 (84%) of the zones with gaps completely filled in, and all 4- and 5-mm gaps filled in. There was no progression of any postoperative gap, no evidence of continuous periacetabular interface radiolucencies, no evidence of lysis, and no revisions for loosening. Although these short-term results are encouraging, further follow-up will be required to assess whether the monoblock design and the low modulus of elasticity of porous tantalum will reduce the incidence of periacetabular stress shielding and occurrence of osteolysis. PMID- 15809958 TI - Third-body wear testing of a highly cross-linked acetabular liner: the effect of large femoral head size in the presence of particulate poly(methyl-methacrylate) debris. AB - The hip simulator wear performance of an electron beam cross-linked and subsequently melted ultrahigh molecular weight polyethylene against femoral heads of 28-, 38-, and 46-mm diameter in the presence of poly(methyl-methacrylate) particulate debris was contrasted with that of conventional polyethylene against a 46-mm diameter head. Over 5 million cycles of testing, the average wear rate of the conventional polyethylene liners was 29.3 +/- 3.0 mg per million cycles. All highly cross-linked components exhibited marked reduction in wear, with the highest wear measuring 0.74 +/- 0.85 mg per million cycles. This study, using a clinically relevant third-body material, showed the electron beam cross-linked material to be far more resistant to this third-body wear than conventional ultrahigh molecular weight polyethylene, even when very large diameter femoral heads were used. PMID- 15809959 TI - Stress risers between two ipsilateral intramedullary stems: a finite-element and biomechanical analysis. AB - Periprosthetic fracture after total joint replacement predominantly occurs at the stem tip. In this study, the effects of gap size, stem stability and cortical thickness between two press-fit, ipsilateral intramedullary stems on the tensile stresses created in the femur were investigated using finite-element models. The findings were confirmed with strain-gauge tests using a composite Sawbone femur. Gap size did not affect the level of stress on the femur. Cortical thickness had an important effect on stress distribution: peak stresses increased as bone cortical thickness decreased. Irrespective of gap size, the tips of loose stems acted as stress risers particularly with thinner cortices; the tips of well-fixed stems, however, did not. PMID- 15809960 TI - Stress fracture of the femoral neck as a complication of total knee arthroplasty. AB - Stress fracture of the hip is a rare complication of total knee arthroplasty that occurs most often in patients in whom a significant deformity of the knee has been corrected, particularly those with poor mobility before surgery. We report 4 cases of ipsilateral fracture of the femoral neck after total knee arthroplasty. PMID- 15809961 TI - Total hip arthroplasty in severe congenital factor VII deficiency: successful use of recombinant activated factor VII for hemostasis. AB - Congenital factor VII (FVII) deficiency is a rare hemorrhagic disorder, and, in affected patients, surgery is likely to cause excessive bleeding. In this article, we report the first case of total hip arthroplasty in a patient with severe congenital FVII deficiency in which recombinant activated factor VII (rFVIIa, NovoSeven) was successfully used to manage bleeds. The patient was a 68 year-old woman with severe congenital FVII deficiency, who suffered from a hemorrhagic arthrosis of her left hip joint. We administered rFVIIa as intermittent bolus infusions both perioperatively and postoperatively; dosing was based on the results of a preoperative rFVIIa challenge test. During surgery and the 3-day postoperative treatment period, we observed normal hemostasis with no excessive bleeding or complications. PMID- 15809962 TI - Bilateral prosthetic knee infection by Campylobacter fetus. AB - We present the first documented case of a bilateral prosthetic knee joint infection with Campylobacter fetus. Our patient's risk factors included age, the presence of prosthetic joints, and potential exposure through his contact with farm animals. It is important to be aware of the possibility of C fetus joint infections in high-risk patients who present with pain after total joint arthroplasty. PMID- 15809964 TI - The stability of the cemented tibial component of total knee arthroplasty: posterior cruciate-retaining versus posterior-stabilized design. PMID- 15809967 TI - Meperidine-induced seizure after revision hip arthroplasty. PMID- 15809968 TI - Isolation and structure elucidation of ishwarol B. AB - Ishwarol B was isolated from the bark oil of Cedrelopsis grevei and its structure elucidated by 1D and 2D NMR spectroscopy. PMID- 15809969 TI - Characterization of monofluorinated polycyclic aromatic compounds by 1H, 13C and 19F NMR spectroscopy. AB - Monofluorinated polycyclic aromatic hydrocarbons (F-PAHs) have attracted much attention in analytical, environmental, toxicological and mechanistic studies because of their physico-chemical properties, which are closely similar to those of the parent PAHs. Because of this, full NMR characterization has become of interest. Complete 1H, 13C and 19F NMR chemical shifts, and also 1J(H,C), (n)J(C,F), (n)J(H,F) and (n)J(H,H) coupling constants, have been assigned for the F-PAHs 1-fluoronaphthalene, 2-fluorofluorene, 5-fluoroacenaphthylene, 2 fluorophenanthrene, 3-fluorophenanthrene, 3-fluorofluoranthene, 1-fluoropyrene, 1 fluorochrysene, 2-fluorochrysene, 3-fluorochrysene and 9 fluorobenzo[k]fluoranthene. To allow comparison with the corresponding parent PAHs, the 1H and 13C chemical shifts of acenaphthylene, phenanthrene, fluoranthene, pyrene and benzo[k]fluoranthene were determined. Chemical shift increments and the effects on the coupling constants from the fluorine substitution are discussed. PMID- 15809970 TI - Analysis of aberrant somatic hypermutation (SHM) in non-Hodgkin's lymphomas of patients with chronic HCV infection. AB - Hepatitis C virus (HCV) and aberrant somatic hypermutation (SHM) have each been suggested to contribute to the development of B-cell non-Hodgkin's lymphoma (NHL). The incidence of PIM-1, PAX-5, RhoH/TTF, and c-MYC mutations in tumour biopsy specimens from 32 HCV-infected B-cell NHL patients was analysed to determine whether the extent of aberrant SHM among these patients differed from that previously reported for HCV-negative B-cell NHL patients. Mutation of PIM-1, PAX-5, RhoH/TTF, and c-MYC was detected in 4 (13%), 5 (16%), 4 (13%), and 4 (13%) of 32 samples, respectively. In HCV-positive B-cell NHL patients, the frequency of aberrant SHM was lower than that already found in HCV-negative B-cell NHL patients. This indicates that, unlike B-cell lymphomas from HCV-negative patients, aberrant SHM may not contribute significantly to malignant transformation in HCV-associated B-cell lymphomas. PMID- 15809971 TI - Expression profiling of murine intestinal adenomas reveals early deregulation of multiple matrix metalloproteinase (Mmp) genes. AB - Initiation of intestinal tumours occurs as a consequence of aberrant Wnt signalling. This causes altered expression of a number of genes which provides the mechanistic basis of neoplastic change in normal epithelium. In order to identify these genes, expression profiles of normal intestinal mucosa and intestinal adenomas from multiple intestinal neoplasia (Min) mice were compared. A total of 116 genes were found to show significant changes in expression in adenomas compared with normal mucosa. Functional classification of these genes clearly identified the biological processes of cellular adhesion and matrix remodelling to be profoundly affected. Notably, three members of the matrix metalloproteinase (Mmp) gene family (Mmp10, Mmp13, and Mmp14) were consistently up-regulated in tumour tissue. To extend these data, expression of 17 Mmp genes was defined using quantitative reverse transcriptase-polymerase chain reaction (Q RT-PCR). Several Mmp genes were profoundly up-regulated and every tumour showed overexpression of at least four Mmp genes. These results underscore the probable importance of interactions between the intestinal epithelium and stroma in early tumour development. Furthermore, the inferred role of Mmps at the adenomatous stage of tumourigenesis suggests that this may represent the optimal therapeutic window for the use of Mmp antagonists as anti-cancer agents. PMID- 15809975 TI - Non-empirical calculations of NMR indirect carbon-carbon coupling constants. Part 12--aliphatic and alicyclic oximes. AB - One-bond carbon-carbon coupling constants were calculated in a series of nine aliphatic and alicyclic oximes at the SOPPA (second-order polarization propagator approach) level in good agreement with the available experimental data, and several unknown couplings were predicted with high reliability. The experimental difference between J(C,C) of the corresponding carbon-carbon bonds in cis and trans orientations to the nitrogen lone pair is very well reproduced at the SOPPA level, and this provides an additional tool in the configurational assignment at the C=N bond in oximes and related systems. PMID- 15809976 TI - Abnormal expression of period 1 (PER1) in endometrial carcinoma. AB - The development of endometrial carcinoma (EC) is a multiple-step process, which includes inactivation of tumour suppressor genes, activation of oncogenes, and disturbance of cancer-related genes. Recent studies have shown that the circadian cycle may influence cancer development and prognosis. In this study, the expression of a circadian gene, PER1, was examined in 35 ECs and paired non tumour tissues by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Expression levels of PER1 were significantly decreased in EC, and mutational analysis of the coding regions, together with methylation analysis of cytosine-phosphate guanosine (CpG) sites in the promoter area, was performed to investigate the possible mechanisms. The analyses detected four single nucleotide polymorphisms in both tumour and non tumour tissues, which had no relationship with the expression of PER1. In the promoter area of the PER1 gene, the CpG sites were methylated in 31.4% of ECs, but in 11.4% of paired non-tumour tissues (p < 0.05). These results suggest that the down-regulation of PER1 expression in EC was partly due to inactivation of the PER1 gene by DNA methylation of the promoter and partly due to other factors. Analysis of the relationships between the expression of PER1, P53, c-MYC, cyclin A, cyclin B, and cyclin D1 showed no definite relationship. These results suggest that down-regulation of the PER1 gene disrupts the circadian rhythm, which may favour the survival of endometrial cancer cells. PMID- 15809977 TI - Differential expression and response to anti-TNFalpha treatment of infiltrating versus resident tissue macrophage subsets in autoimmune arthritis. AB - Synovial macrophages play a pivotal role in the pathogenesis of chronic autoimmune arthritis by contributing to local inflammation and tissue damage and are therefore a primary target for therapeutic intervention. The aim of the present study was to investigate in more detail the relative contribution of different synovial macrophage subsets with potentially different inflammatory or anti-inflammatory functions by analysing the two most frequent forms of human autoimmune arthritis, spondyloarthropathy (SpA) and rheumatoid arthritis (RA). Both infiltrating macrophages from peripheral blood expressing myeloid-related proteins (MRP) 8 and 14, and resident tissue macrophages expressing CD163 were abundant in inflamed synovium. Whereas the global number of synovial macrophages was similar in both diseases, infiltrating macrophages were increased in the RA lining layer in contrast with resident tissue macrophages, which were more frequently observed in SpA. Soluble MRP8/MRP14 complexes, which were secreted locally in the joint during the infiltration process, were increased in the serum of arthritis patients and, in contrast with soluble CD163 shed from resident tissue macrophages, correlated well with global inflammatory parameters. Treatment in vivo with anti-TNFalpha had a rapid and pronounced effect on the infiltration of MRP-positive macrophages into tissues, as evidenced by histopathological analysis and serum MRP8/MRP14 levels. Taken together, these data support an important role for infiltrating versus resident tissue macrophages in human autoimmune synovitis and indicate that macrophage products such as soluble MRP8/MRP14 complexes are valuable biomarkers for the experimental and clinical monitoring of specific disease mechanisms in vivo. PMID- 15809978 TI - Total assignments of 1H and 13C NMR spectra of isocatalpanol and a derivative of tecomaquinone. AB - Isocatalpanol and tecomaquinone I were obtained from roots of Lippia sidoides, a medicinal plant from northeast Brazil. Reduction of tecomaquinone I with NaBH4 yielded a new derivative. Structural elucidation was done on the basis of spectral data, mainly by high-field NMR and electron ionization mass spectrometry. PMID- 15809979 TI - Comparison of the phytoestrogen trans-resveratrol (3,4',5-trihydroxystilbene) structures from x-ray diffraction and solution NMR. AB - The NMR-derived solution structure of trans-3,4',5-trihydroxystilbene (resveratrol) was compared with two recent literature crystal x-ray structures, resveratrol in complex with human transthyretin (TTR-RES) from 1DVS.pdb and resveratrol bound to chalcone synthase (CHS-RES) from 1CGZ.pdb. 1H and 13C NMR spectra of resveratrol were acquired in DMSO-d6. Assignments were obtained from an analysis of DQF-COSY, TOCSY, DEPT, HMQC/HSQC, HMBC and INADEQUATE NMR spectra. Past 1H and 13C NMR literature assignments are corrected. The dihedral angle 2-1 1'-2' provides an indication of the relative spatial orientation of the two phenolic rings. Values of 1.62, - 54.10 and 12.6 +/- 1.1 degrees were found for the 1DVS.pdb, 1CGZ.pdb and NMR resveratrol structures, respectively. The 1DVS.pdb resveratrol structure is 'flat' with the two phenolic rings along the same plane. The 1CGZ.pdb structure has these two rings almost orthogonal to each other, and the NMR structure has these two rings much closer to being along the same plane. The angles 1-alpha--alpha' and 1'-alpha'--alpha are along the same trace and of similar magnitude for the 1CGZ.pdb and NMR resveratrol structures. For the 1DVS.pdb resveratrol structure, these angles are about 7-10 degrees greater, with alpha and alpha' being 180 degrees out-of-phase from the other two structures. The alpha rings did not overlap, with the NMR result representing a 'median model' of the two x-ray structures. PMID- 15809980 TI - Structure elucidation and complete NMR spectral assignments of a new taxane glycoside from the needles of Taxus cuspidata. AB - A new taxane glycoside was isolated from a methanol extract of the needles of Taxus cuspidata. The structure was established as 2alpha,9alpha,10beta triacetoxytaxa-4(20),11-dien-13-one-5alpha-O-beta-D-glucopyranoside (1) on the basis of 1D- and 2D NMR and high-resolution fast atom bombardment mass spectral analyses. PMID- 15809981 TI - Comparison of 17beta-estradiol structures from x-ray diffraction and solution NMR. AB - The NMR-derived structure of estrogen (17beta-estradiol, E2), the drug of choice for postmenopausal women, was compared with a recent literature crystal x-ray structure of Fab-bound E2. 1H and 13C NMR spectra of E2 were acquired in DMSO-d6. Assignments were obtained from an analysis of DQF-COSY, TOCSY, HETCOR, HMQC and HMBC 2D NMR spectra. The 1H and 13C NMR assignments are the first reported for E2 in DMSO-d6. Two solution structures, S1 and S2, were obtained with molecular modeling using NOE constraints. S1 overlaps with the crystal structure for all rings. S2 shows prominent differences in the C-ring (C9--C11--C12--C13) segment, which deviates from a chair conformation, and excellent overlap in the A-, B- and D-rings of E2. The C-ring in S2 adopts a boat conformation as opposed to a chair conformation in the x-ray and S1 structures. The S2 structure is about 6 degrees more twisted than the bound x-ray and S1 models. The S1, S2 and x-ray structures had ring bowing values of 10.1 +/- 0.3, 11 +/- 1 and 10.37 degrees , respectively. Of the 100 solution conformers generated, 83 had S1 conformation and 17 had S2 conformation, with average internal energies of 112 +/- 2 and 141 +/- 2 kcal mol(-1), respectively. The 100 S1- and S2- derived conformers showed a r.m.s.d. of 0.72 A for all atoms. The x-ray, S1 and S2 C18--O17 distances were 2.93, 2.92 +/- 0.01 and 2.93 +/- 0.01 A, respectively, and the O3--O17 distances were 11.06, 11.18 +/- 0.12, and 10.89 +/- 0.05 A, respectively. PMID- 15809982 TI - Characterization of two minor saponins from Cordia piauhiensis by 1H and 13C NMR spectroscopy. AB - A careful NMR analysis with full assignment of the 1H and 13C spectral data for two minor saponins isolated from stems of Cordia piauhiensis is reported. These saponins were isolated by high-performance liquid chromatography and characterized as 3beta-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D glucopyranosyl]pomolic acid 28-O-[beta-D-glucopyranosyl-(1 --> 6)-beta-D glucopyranosyl] ester (1) and 3beta-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D glucopyranosyl]oleanolic acid 28-O-[beta-D-xylopyranosyl-(1 --> 2)-beta-D glucopyranosyl-(1 --> 6)-beta-D-glucopyranosyl] ester (2). Their structures were established using a combination of 1D and 2D (1H, 1H-COSY, TOCSY, NOESY, gs-HMQC and gs-HMBC) NMR techniques, electrospray ionization mass spectrometry and chemical evidence. PMID- 15809983 TI - 1H and 13C NMR data to aid the identification and quantification of residual solvents by NMR spectroscopy. AB - We present reference data and a javascript web page which allow the rapid identification and quantification of residual solvents by NMR. The data encompass all of the ICH-prescribed solvents and were obtained for a number of NMR solvents. We also present an example of its application. PMID- 15809984 TI - Atom-type assignment in molecules and clusters by perturbation theory-A complement to X-ray structure analysis. AB - An approach to distinguish elements with similar atomic numbers in molecules and clusters is presented and applied to experimentally synthesized and structurally characterized mixed-metallic compounds. By first treating a homogenized reference system constructed from the original compound and applying first-order perturbation theory it is possible to find the most stable distribution of the atom types to the atomic sites in a very efficient way. This work is focused on the appropriate choice of homogenized reference systems and on applications treating experimentally synthesized compounds. With these examples is shown that the method is a helpful complement to X-ray crystal structure analysis. PMID- 15809985 TI - Synthesis and study on 2,4,6,8-tetraaryl- 3,7-diazabicyclo[3.3.1]nonanes and their derivatives. AB - 1-Carbethoxy-2,4,6,8-tetraaryl-3,7-diazabicyclo[3.3.1] nonan-9-ones (1, 2) were synthesized and their 1H and 13C NMR data are reported. Chemical shifts and spectral assignments for 2,4,6,8-tetrakis(4-chlorophenyl)-3,7 diazabicyclo[3.3.1]nonan-9-one (3), 2,4,6,8-tetraphenyl-3-thia-7 azabicyclo[3.3.1]nonan-9-one (4) and 2,4,6,8-tetraaryl-3,7 diazabicyclo[3.3.1]nonanes (5-7) are also included. PMID- 15809986 TI - The structure of acrolein in a liquid crystal phase. AB - The (1)H NMR spectrum of a sample of acrolein dissolved in the nematic liquid crystal phase I52 has been analysed to yield 18 dipolar couplings between all the magnetic nuclei in the molecule; moreover, the (13)C and (13)C{(1)H} NMR spectra of a sample of acrolein in CDCl(3) were recorded and analysed to determine the indirect J(ij) couplings. The data were used to obtain the relative positions of the carbon and hydrogen atoms, assuming that these are independent of the conformations generated by rotation around the C--C bond through an angle phi, and to obtain a probability distribution P(phi). It has been found that in the liquid phase, the distribution is a maximum at the trans form whereas the abundance of the cis form is significantly smaller compared with that found by microwave spectroscopy or high level quantum mechanical calculations. Such calculations produced also a suitable force field needed to develop suitable strategies for vibrational correction procedure in the case of flexible molecules. PMID- 15809987 TI - Rhodium-catalyzed nondecarbonylative addition reaction of ClCOCOOC2H5 to alkynes. AB - Addition of ethoxalyl chloride (ClCOCOOEt) to terminal alkynes at 60 degrees C in the presence of a rhodium(I)-phosphine complex catalyst chosen from a wide range affords 4-chloro-2-oxo-3-alkenoates regio- and stereoselectively. Functional groups such as chloro, cyano, alkoxy, siloxy, and hydroxy are tolerated. The oxidative addition of ethoxalyl chloride to [RhCl(CO)(PR(3))(2)] proceeds readily at 60 degrees C or room temperature and gives [RhCl(2)(COCOOEt)(CO)(PR(3))(2)] (PR(3) = PPh(2)Me, PPhMe(2), PMe(3)) complexes in high yields. The structure of [RhCl(2)(COCOOEt)(CO)(PPh(2)Me)(2)] was confirmed by X-ray crystallography. Thermolysis of these ethoxalyl complexes has revealed that those ligated by more electron-donating phosphines are fairly stable against decarbonylation and reductive elimination. [RhCl(2)(COCOOEt)(CO)(PPh(2)Me)(2)] reacts with 1-octyne at 60 degrees C to form ethyl 4-chloro-2-oxo-3-decenoate. The catalysis is therefore proposed to proceed by oxidative addition of ethoxalyl chloride, insertion of an alkyne into the Cl--Rh bond of the resulting intermediate, and reductive elimination of alkenyl-COCOOEt. PMID- 15809988 TI - Intervalence charge transfer (IVCT) in ruthenium dinuclear and trinuclear assemblies containing the bridging ligand HAT {1,4,5,8,9,12-hexaazatriphenylene}. AB - The IVCT characteristics of the mixed-valence forms of the dinuclear [{Ru(bpy)(2)}(2)(mu-hat)](n+) and the trinuclear [{Ru(bpy)(2)}(3)(mu-hat)](n+) species {HAT = 1,4,5,8,9,12-hexaazatriphenylene; bpy = 2,2'-bipyridine} show a marked dependence on the nuclearity, and in the trinuclear case on the extent of oxidation. Small differences are also found between the diastereoisomers of the dinuclear complex {meso (DeltaLambda) and rac (DeltaDelta/LambdaLambda)}, and between the homochiral (Delta(3)/Lambda(3)) and heterochiral (Delta(2)Lambda/Lambda(2)Delta) diastereoisomers of the trinuclear case. The strong metal-metal interactions result in unusual spectroscopic and electrochemical properties of the singly-oxidised (+7) and doubly-oxidised (+8) trinuclear mixed-valence species. A qualitative localised bonding description based on the geometrical properties of the dpi(Ru(II/III)) orbitals is invoked to explain the IVCT behaviour in the di- and trinuclear systems. PMID- 15809989 TI - Discovery of powerful uranyl ligands from efficient synthesis and screening. AB - New tripodal gem-(bis-phosphonates) uranophiles were discovered by a screening method that allowed for the selection of ligands with strong uranyl-binding properties in a convenient microtiter-plate format. The method is based on competitive uranium binding by using Sulfochlorophenol S as chromogenic chelate. This dye compound was found to present high uranyl complexation properties and allowed to highlight ligands presenting association constants for UO(2+)(2) up to 10(18) at pH 7.4 and 10(20) at pH 9. A collection of 40 known ligands including polycarboxylate, hydroxamate, catecholate, hydroxypyridonate and hydroxyquinoline derivatives was tested. Also screened was a combinatorial library prepared from seven amine scaffolds and eight acrylates bearing diverse chelating moieties. Among these 96 tested candidates, a tripod derivative bearing gem-bis phosphonates moieties was found to present the highest complexation properties over a wide range of pH and was further studied. PMID- 15809990 TI - Early diagnosis remains the most reliable way to cure children with melanoma. PMID- 15809991 TI - Anatomical alterations of the subthalamic nucleus in relation to age: a postmortem study. AB - The subthalamic nucleus (STN) is currently the preferred target for chronic electrical high-frequency stimulation in Parkinson's disease. Anatomical determination of the exact position of the STN in the individual patient, using magnetic resonance imaging, remains cumbersome, whereas calculation of the target using a stereotactic atlas bypasses patient interindividual variations in the exact delineation of the STN. The aim of this study was to demonstrate variations in shape and position of the STN during life. In this anatomopathological study, a method was applied to localize the STN in reference to the anterior commissure posterior commissure line (AC-PC line) in 12 postmortem brains of patients who died of non-neurological diseases. Their age varied from 29 to 84 years. Centers and borders of the STN were macroscopically measured in three spatial orthogonal planes in relation to the AC-PC line, and verified by light microscopy. The AC-PC distance remains almost constant during life (24.4 mm; SD 3.58). With increasing age, the center of the STN tends to move 3.9 mm cranially, 2.6 mm laterally, and 0.2 mm anteriorly. This last result also differs from the position mentioned in the stereotactic brain atlases. The form of the STN also changes. During life, the STN becomes wider in the mediolateral direction and smaller in the superior inferior and anterior-posterior direction. The shape and spatial position of the STN also change during life. These changes should be taken into account during target determination in deep brain stimulation procedures in Parkinson's disease. PMID- 15809992 TI - A Markov model approach shows a large variation in the length of S phase in MCF-7 breast cancer cells. AB - BACKGROUND: The potential doubling time of a tumor has been suggested to be a measurement of tumor aggressiveness; therefore, it is of interest to find reliable methods to estimate this time. Because of variability in length of the various cell cycle phases, stochastic modeling of the cell cycle might be a suitable approach. METHODS: The relative movement curve and the DNA synthesis time were estimated by using local polynomial regression methods. Further, the rate of nucleotide incorporation was estimated by using a Markov pure birth process with one absorbing state to model the progression of the DNA distribution through S phase. RESULTS: An estimate of the DNA synthesis time, with confidence intervals, was obtained from the relative movement curve. The Markov approach provided an estimate of the distribution of the time to complete S phase given the initial distribution. Using the Markov approach we also made an estimate of the mean number of active replicons during S phase. CONCLUSIONS: A Markov pure birth process has shown to be useful to model the progression of cells through S phase and to increase knowledge about the variability in the length of S phase and a large variation is shown. PMID- 15809993 TI - Split hand malformation, hypospadias, microphthalmia, distinctive face and short stature in two brothers suggest a new syndrome. AB - Split hand/foot malformation (SHFM) is a genetically heterogeneous limb malformation that may be isolated or associated with other malformations. More than 50 recognizable entities with SHFM have been described and at least 5 mapped genetic loci have been implicated. Two brothers with intrauterine growth retardation, short stature, distinctive face, microphthalmia, genital anomalies, and SHFM are described. Molecular analyses of TP63, HOXA13, and HOXD13 genes were normal. We propose this pattern to be a newly recognized SHFM syndrome. PMID- 15809994 TI - Novel movement disorder of the lower lip: is it epilepsia partialis continua? Clues from a secondary case. AB - A 28-year-old woman developed an acute-onset novel movement disorder of the lower lip mimicking focal dystonia. Investigations showed it to be a presentation of epilepsia partialis continua occurring in association with agenesis of the corpus callosum. It responded favorably to anti-epileptic drug therapy. Recently, Kleopa and Kyriakides reported on 4 patients who developed sudden-onset movement disorder characterized by a tonic sustained, lateral and outward protrusion of half of the lower lip. They failed to find any causative factors, despite extensive investigation. Treatment with anticholinergics, clonazepam, and botulinum toxin injection failed to improve the movement disorder. I present an additional case of similar focal movement disorder occurring in the presence of agenesis of the corpus callosum. A scalp electroencephalogram revealed focal epileptic activity, and the movement disorder responded favorably to treatment with antiepileptic drugs. PMID- 15809995 TI - Rimantadine in Parkinson's disease patients experiencing peripheral adverse effects from amantadine: report of a case series. AB - We report our experience with 7 consecutive patients with Parkinson's disease (PD) who received rimantadine (the alpha-methyl derivative of amantadine) in substitution of amantadine due to peripheral side effects (lower limb edema, livedo reticularis). Mean age was 67.3 +/- 5.9 years, the mean disease duration was 13 +/- 6.3 years, and mean Hoehn and Yahr stage was 2.2 +/- 0.4. A total of 3 patients experienced marked improvement of edema, and 1 patient experienced marked improvement of livedo reticularis. Only 1 of the 7 patients reported significant loss of motor benefit when amantadine was replaced with rimantadine. Our results demonstrate that rimantadine may be considered as an alternative to amantadine in patients experiencing amantadine-induced peripheral side effects. PMID- 15809996 TI - An unusual reciprocal translocation detected by subtelomeric FISH: interstitial and not terminal. AB - An 11-month-old boy with a pattern of dysmorphic signs, an atrial septal defect, right inguinal hernia, bilateral undescended testes, bilateral urinary reflux, right renal dysplasia, and developmental delay had an abnormal chromosome 11 with additional material of unknown origin attached to the long arm in his karyotype. The paternal karyotype was normal 46,XY, while the mother's karyotype was 46,XX,t(2;11)(q35;q24.2). Thus, a reciprocal terminal exchange was assumed resulting in duplication of distal 2q material and a small subterminal 11q deletion. However, application of subtelomeric fluorescence in situ hybridization (FISH) probes indicated that the translocation was not a terminal reciprocal exchange, but was interstitial at least for one of the chromosomes, which would be highly unusual since most interstitial translocations are non-reciprocal. Based on the results of FISH and microsatellite marker examinations, the designation of the breakpoints and thus of the deleted and duplicated segments had to be revised. The findings have implications for karyotype-phenotype correlation. PMID- 15809997 TI - Attention-deficit/hyperactivity disorder (ADHD) and variable clinical expression of Aarskog-Scott syndrome due to a novel FGD1 gene mutation (R408Q). AB - Mutations of the FGD1 gene are responsible for a significant proportion of patients with Aarskog-Scott syndrome (AAS), an X-linked disorder characterized by short stature, brachydactyly, urogenital abnormalities, and a typical dysmorphic facial appearance. Although mental retardation does not occur significantly in AAS, this condition has been described associated with various degrees of mental impairment and/or behavioral disorders in some patients. In particular, attention deficit hyperactivity disorder (ADHD) is reported as a common characteristic of AAS. However, AAS/ADHD reported patients have been only clinically described, and diagnosis never has been confirmed on molecular basis. We present here a unique case of a 16-years-old patient presenting with ADHD, lower intelligence quotient, and dysmorphic features. Although the clinical features were not completely typical of AAS, genetic analysis demonstrated a novel FGD1 missense mutation (R408Q). The case we report confirms the highly variable expressivity of AAS and first documents that the FGD1 gene may play a role in ADHD susceptibility. We suggest that FGD1 analysis may be adequate in ADHD patients who exhibit dysmorphic features suggestive of AAS, also in the absence of the full phenotypical spectrum. PMID- 15809998 TI - Familial recurrence rates and genetic models of multiple sclerosis. AB - Susceptibility to multiple sclerosis (MS) is determined by both inherited and non inherited factors. The importance of genetic factors is demonstrated by the increased risk of disease in relatives of MS patients. Our objective was to determine the implications of the observed familial recurrence risks for the genetic basis of MS. We developed a computer program which calculates recurrence risks for monozygotic (MZ) twins, siblings, and second degree relatives, and used it to calculate recurrence risks for a wide variety of genetic models. We investigated models with different numbers of genes, different patterns of interaction between the genes, and dominant or recessive inheritance. The models that best reproduced the observed values had multiple loci with strongly synergistic interaction and autosomal dominant (AD) inheritance. At least six loci were required, and we found no upper limit on the number of loci. Models with genetic heterogeneity, where only a fraction of the risk loci are required for disease, are possible. In models with large numbers of loci the "abnormal" alleles conferring risk of disease are the most common allele. We conclude that a variety of genetic models with multiple genes, dominant inheritance, and synergistic interaction between risk genes are consistent with the observed familial recurrence rates in MS. PMID- 15809999 TI - Familial variable expression of dilated cardiomyopathy in Alstrom syndrome: a report of four sibs. AB - Alstrom syndrome is an autosomal recessive disorder comprised of progressive vision loss (nystagmus, photophobia, and pigmentary retinopathy), progressive sensorineural hearing loss, morbid obesity, male hypogonadism, insulin resistant diabetes, renal failure, and dilated cardiomyopathy. We report on four sibs with Alstrom syndrome with intra-familial variability in onset, severity, and spectrum of manifestations; the most serious manifestation being dilated cardiomyopathy. This report emphasizes the difficulty of recognizing this constellation of symptoms as Alstrom syndrome at an early age, the seriousness of cardiac involvement, and the intra-familial variability of phenotypic expression. PMID- 15810000 TI - Physician exposure to and attitudes toward advertisements for genetic tests for inherited cancer susceptibility. AB - Commercial marketing materials may serve as a source of information for physicians about genetic testing for inherited cancer susceptibility (GTICS) in addition to medical guidelines, continuing education, and journal articles. The primary purposes of this study were to: (1) determine the percentage of physicians who received advertisements for GTICS early in the diffusion of commercial GTICS (1999-2000); (2) assess associated characteristics; and (3) measure the perceived importance of commercial advertisements and promotions in physicians' decisions to recommend testing to patients. A nationally representative, stratified random sample of 1,251 physicians from the American Medical Association (AMA) Physician Masterfile completed a 15-20 min mixed mode questionnaire that assessed specialty, previous use of genetic tests, practice characteristics, age, and receipt of advertising materials (response rate = 71%). Overall, 27.4% (n = 426) had received advertisements. In multivariate analysis, factors associated with receipt of advertisements included: specialties in obstetrics/gynecology, oncology, or gastroenterology; past GTICS use, and age 50+. One of four felt that advertisements would be important in their decision to recommend GTICS. Study results indicate that physicians, particularly in oncology, obstetrics/gynecology, and gastroenterology, began receiving GTICS advertisements commensurate with the early diffusion of commercially available tests into clinical practice. At that time, one-quarter of the physicians considered advertisements to play an important role in their clinical decision making, suggesting attention to other sources of information and additional factors. PMID- 15810001 TI - A novel MGP mutation in a consanguineous family: review of the clinical and molecular characteristics of Keutel syndrome. AB - Keutel syndrome (KS) [OMIM 245150] is a rare autosomal recessive condition, characterized by abnormal cartilage calcification. Mutations in the matrix Gla protein gene (MGP) have been previously reported in three unrelated KS families. MGP is an extracellular matrix protein that acts as a calcification inhibitor by repressing bone morphogenetic protein 2 (BMP2). Loss-of-function mutations of MGP result in abnormal calcification of the soft tissues, a cardinal feature of KS. We report the fourth MGP mutation (IVS2 + 1G > A) in a consanguineous Arab family, which results in the loss of the consensus donor splice site at the exon 2-intron 2 junction. In addition to the typical manifestations, we observed abnormalities in the white matter of the brain, optic nerve atrophy, and mid dermal elastolysis in the affected individuals of this family. This report broadens the clinical phenotype observed in patients with KS. The effect of the IVS2 + 1G > A mutation is consistent with the previously reported loss-of function mutations of MGP. PMID- 15810002 TI - Phenotypes with GATA4 or NKX2.5 mutations in familial atrial septal defect. AB - Recently, GATA4 and NKX2.5 were reported as the disease genes of atrial septal defect (ASD) but the relationship between the locations of their mutations and phenotypes is not clear. We analyzed GATA4 and NKX2.5 mutations in 16 familial ASD cases, including four probands with atrioventricular conduction disturbance (AV block) and two with pulmonary stenosis (PS), by PCR and direct sequencing, and examined their phenotypes clinically. Five mutations, including two GATA4 and three NKX2.5 mutations, were identified in 31.3% of the probands with ASD, and three of them were novel. The two GATA4 mutations in the probands without AV block were S52F and E359Xfs (c.1075delG) that was reported previously, and three NKX2.5 mutations in the probands with AV block were A88Xfs (c.262delG), R190C, and T178M. Additionally, we observed some remarkable phenotypes, i.e., dextrocardia with E359Xfs (c.1075delG) and cribriform type ASD with R190C, both of which are expected to be clues for further investigations. Furthermore, progressive, most severe AV block was closely related with a missense mutation in a homeodomain or with a nonsense/frame-shift mutation of NKX2.5 for which classification has not been clearly proposed. This pinpoints essential sites of NKX2.5 in the development of the conduction system. PMID- 15810003 TI - Molecular characterization of a 14q deletion in a boy with features of Holt-Oram syndrome. AB - Holt-Oram syndrome, the major "heart-hand" syndrome is defined by the association of radial defects or triphalangeal thumbs and septal heart defects. The transmission is autosomal dominant and the causative gene has been shown to be TBX5, located on 12q24.1, which encodes a transcription factor. Genetic heterogeneity has been suggested by several reports. We identified a 14(q23.3 approximately 24.2q31.1) deletion in a boy presenting severe bilateral asymmetrical radial aplasia, congenital heart defects, and developmental delay. This deletion, whose size could be estimated to be 9.6-13.7 Mb, was shown to be inherited via his mother's interchromosomal insertion. This is the second report of a chromosome 14 interstitial deletion associated with clinical features of Holt-Oram syndrome. These observations suggest the existence of a new "heart hand" locus on chromosome 14q. PMID- 15810004 TI - Subtelomere deletions and translocations are frequently familial. AB - In recent years, strategies have been developed to investigate the possible role of chromosomal subtelomere regions in genetic disorders. The present study was to determine the incidence of familial subtelomeric abnormalities among individuals with developmental delay, idiopathic mental retardation, or non-specific congenital abnormalities. A review was conducted for patients and their relatives on whom subtelomeric DNA fluorescence in situ hybridization (telo-FISH) studies were performed. Patients were identified through a search of the Mayo Genetics System (MGS) database. Of 2,170 consecutive telo-FISH index case studies completed in our laboratory between January 2002 and December 2003, 121 or 5.6% had abnormalities of the subtelomere region. The present report includes 18 other abnormal index cases seen prior to 2002 to yield a total of 139 abnormal index cases. This represents 71 index patients with deletions, 53 index patients with derivative chromosomes, and 15 index patients with balanced rearrangements. A familial abnormality was identified in 29 (51.8%) of 56 families in whom parents and/or sibs were available for testing. Among 28 patients with deletions, 9 (32%) had an inherited deletion, whereas 19 (68%) were de novo. Family members of 20 index patients with derivative chromosomes were tested. Of these, 13 (65%) patients inherited the abnormality from a parent (12 from a parent who had a balanced translocation and 1 from a parent with the same abnormality), while 7 (35%) apparently arose de novo. Seven (88%) of 8 with balanced translocations inherited the translocation from one parent. The most common familial abnormalities involved 8pter deletion or rearrangement. The incidence of familial subtelomeric abnormalities is significantly high making parental telo-FISH studies an essential part of the investigation of patients with subtelomeric chromosome abnormalities. PMID- 15810005 TI - New method for estimation of adult skeletal age at death from the morphology of the auricular surface of the ilium. AB - A new method for estimating skeletal age at death from the morphology of the auricular surface of the ilium is presented. It uses a multiple regression analysis with dummy variables, and is based on the examination of 700 modern Japanese skeletal remains with age records. The observer using this method needs only to check for the presence or absence of nine (for a male) or seven (for a female) features on the auricular surface and to select the parameter estimates of each feature, calculated by multiple regression analysis with dummy variables. The observer can obtain an estimated age from the sum of parameter estimates. It is shown that a fine granular texture of the auricular surface is typical of younger individuals, whereas a heavily porous texture is characteristic of older individuals, and that both of these features are very useful for estimating age. Our method is shown here to be more accurate than other methods, especially in the older age ranges. Since the auricular surface allows more expedient observations than other parts of the skeleton, this new method can be expected to improve the overall accuracy of estimating skeletal age at death. PMID- 15810006 TI - Examination of increased annual range of a Tana mangabey (Cercocebus galeritus) group. AB - A group of Tana mangabeys (Cercocebus galeritus) has greatly expanded its annual range over three decades: from 17.25 hectares (ha) in 1974 (Homewood [1976] Ph.D. dissertation, University College London) and 18.75 ha in 1988-1989 (Kinnaird [1990a] Ph.D. dissertation, University of Florida) to 46.75 ha in 2000-2001. Utilizing 5-min mapping samples, phenological samples, vegetation data, and statistical and descriptive comparisons with previous data, three hypotheses concerning this range increase were postulated. The hypothesis with the strongest support is an increase in intragroup feeding competition, which was measured by a decrease in fruit availability per hectare and an increase in group size. Fruit biomass estimates of four species individually and of eight species combined showed significant decreases between earlier findings (Kinnaird [1990] Ph.D. dissertation, University of Florida) and this study. Group size decreased from 36 in 1974 to 17 in 1988-1989, and is now 50 individuals. There is no support for the hypotheses that the group is no longer constrained in home range size by neighboring groups or by the extent of forest cover. The changes in group and home range size are also discussed in the context of historical forest loss in the lower Tana River. The ability of the Tana mangabey to increase its home range, especially by traveling through nonforest habitat, is an important aspect of its ecological flexibility in a fragmented and threatened habitat. PMID- 15810007 TI - Statin drugs and congenital anomalies. PMID- 15810008 TI - Craniofacial defects of blastogenesis: duplication of pituitary with cleft palate and orophgaryngeal tumors. AB - Duplications of organs and/or tissues are rare in morphogenesis and have frequently been attributed to incomplete twinning. To further elucidate the phenotypes associated with organ duplications, we present three infants with duplication of the pituitary gland (DPG). A review of previously reported cases with DPG showed that the commonest additional findings were hypothalamic enlargement, a broad or duplicated sella, cleft palate, hypertelorism, oropharyngeal tumors, agenesis or hypoplasia of the corpus callosum, and abnormalities of vertebrae. DPG and additional malformations constitute a distinct and recognizable pattern of anomalies, which may constitute a polytopic field defect due to splitting of the notochord. However, the precise inductive mechanism resulting in DPG remains unknown. PMID- 15810009 TI - External forces on the limbs of jumping lemurs at takeoff and landing. AB - Ground reaction forces were recorded for jumps of three individuals each of Lemur catta and Eulemur fulvus. Animals jumped back and forth between a ground-mounted force plate and a 0.5-m elevated platform, covering horizontal distances of 0.5-2 m. In total, 190 takeoffs and 263 landings were collected. Animals typically jumped from a run up and into a run out, during which they gained or into which they carried horizontal impulse. Correspondingly, vertical impulses dominated takeoffs and landings. Peak forces were moderate in magnitude and not much higher than forces reported for quadrupedal gaits. This is in contrast to the forces for standing jumps of specialized leapers that considerably exceed forces associated with quadrupedal gaits. Force magnitudes for the lemur jumps are more comparable to peak forces reported for other quadrupeds performing running jumps. Takeoffs are characterized by higher hindlimb than forelimb peak forces and impulses. L. catta typically landed with the hindlimbs making first contact, and the hindlimb forces and impulses were higher than the forelimb forces and impulses at landing. E. fulvus typically landed with the forelimbs striking first and also bearing the higher forces. This pattern does not fully conform to the paradigm of primate limb force distribution, with higher hindlimb than forelimb forces. However, the absolute highest forces in E. fulvus also occur at the hindlimbs, during acceleration for takeoff. PMID- 15810010 TI - Midline raphe, sternal cleft, and other midline abnormalities: a new dominant syndrome? AB - Sternal fusion defects are malformations that often present as an isolated finding. An association with multiple malformations has been reported, in particular with midline raphe and craniofacial hemangiomas and as part of the pentalogy of Cantrell. Most syndromic cases were sporadic with a few families reported with recurrence in sibs. We describe a mother and two daughters with midline raphe and sternal defects. Affected members also had double central incisors, congenital heart defect, neck webbing, bicornuate uterus and minor anomalies including long face with hypotelorism. None of the three affected relatives had hemangiomas. The manifestations of these patients do not fit any previously described condition, and we propose they represent a new syndrome. This family is also important as it points to a possible genetic cause for at least some cases of this disorder of the ventral midline. PMID- 15810011 TI - Enhanced susceptibility of S-100B transgenic mice to neuroinflammation and neuronal dysfunction induced by intracerebroventricular infusion of human beta amyloid. AB - S-100B is an astrocyte-derived protein that is increased in focal areas of the brain most severely affected by neuropathological changes in Alzheimer's disease (AD). Cell-based and clinical studies have implicated S-100B in progression of a pathologic, glial-mediated pro-inflammatory state in the CNS. However, the relationship between S-100B levels and susceptibility to AD-relevant neuroinflammation and neuronal dysfunction in vivo has not been determined. To test the hypothesis that overexpression of S-100B increases vulnerability to beta amyloid (Abeta)-induced damage, we used S-100B-overexpressing transgenic (Tg) and S-100B knockout (KO) mice in a mouse model that involves intracerebroventricular infusion of human oligomeric Abeta1-42. This model mimics many features of AD, including robust neuroinflammation, Abeta plaques, synaptic damage and neuronal loss in the hippocampus. S-100B Tg, KO, and wild-type (WT) mice were infused with Abeta for 28 days, sacrificed at 60 days, and hippocampal endpoints analyzed. We found that Tg mice showed increased vulnerability to Abeta-induced neuropathology relative to either WT or KO mice. Specifically, Tg mice exhibited enhanced glial activation and neuroinflammation, increased nitrotyrosine staining (a marker of glial-induced neuronal damage), and more pronounced loss of synaptic markers. Interestingly, Tg mice showed no significant differences in Abeta plaque burden compared with WT or KO mice, suggesting that, as in the human situation, the severity of neuronal dysfunction did not correlate with amyloid deposition. Our data are consistent with a model in which S-100B overexpression in AD enhances glial activation and leads to an augmented neuroinflammatory process that increases the severity of neuropathologic sequelae. PMID- 15810012 TI - Occupation, exposure to polycyclic aromatic hydrocarbons and laryngeal cancer risk. AB - Primary risk factors for laryngeal cancer are smoking and alcohol. The relevance of occupational exposures in the etiology of laryngeal cancer is not yet clarified. Some studies have suggested various occupational agents as additional causal risk factors. A population-based case-control study 1:3 frequency matched by age and gender on laryngeal cancer was carried out in southwest Germany with 257 cases (236 males and 21 females between the ages of 37-80, histologically confirmed and diagnosed between January 5, 1998 and December 31, 2000) and 769 population controls (702 males, 67 females). Occupational exposures and other risk factors were obtained with face-to-face interviews using a detailed standardized questionnaire. The complete individual work history was assessed. A detailed assessment of work conditions was obtained by job-specific questionnaires for selected jobs known to be associated with exposure to potential carcinogens. A specific substance list was used as second method for exposure assessment. Blood samples were taken from all individuals for genotype analysis. A strong effect of polycyclic aromatic hydrocarbons exposure on laryngeal cancer risk after adjustment for smoking and alcohol (odds ratio [OR] = 5.2, 95% confidence interval [CI] = 1.6-17.1) was observed for concordant exposure classified with both methods, and a clear dose-response (p < 0.01 for linear trend) for exposure duration. Our findings are supported by risks associated with occupational groups in which this exposure is a priori considered likely. A differential effect by glutathione-S-transferases-M1 genotype was found, however, small numbers do not allow firm conclusions on effect modification. Our study contributes to classifying polycyclic aromatic hydrocarbons as a risk factor for laryngeal cancer. PMID- 15810013 TI - Amino acid recognition by Venus flytrap domains is encoded in an 8-residue motif. AB - A motif foramino acid recognition by proteins or domains of the periplasmic binding protein-like I superfamily has been identified. An initial pattern of 5 residues was based on a multiple sequence alignment of selected proteins of that fold family and on common structural features observed in the crystal structure of some members of the family [leucine isoleucine valine binding protein (LIVBP), leucine binding protein (LBP), and metabotropic glutamate receptor type 1 (mGlu1R) amino terminal domain)]. This pattern was used against the PIR-NREF sequence database and further refined to retrieve all sequences of proteins that belong to the family and eliminate those that do not belong to it. A motif of 8 residues was finally selected to build up the general signature. A total of 232 sequences were retrieved. They were found to belong to only three families of proteins: bacterial periplasmic binding proteins (PBP, 71 sequences), family 3 (or C) of G-protein coupled receptor (GPCR) (146 sequences), and plant putative ionotropic glutamate receptors (iGluR, 15 sequences). PBPs are known to adopt a bilobate structure also named Venus flytrap domain, or LIVBP domain in the present case. Family 3/C GPCRs are also known to hold such a domain. However, for plant iGluRs, it was previously detected by classical similarity searches but not specifically described. Thus plant iGluRs carry two Venus flytrap domains, one that binds glutamate and an additional one that would be a modulatory LIVBP domain. In some cases, the modulator binding to that domain would be an amino acid. PMID- 15810014 TI - Enzymatic incorporation of orthogonally reactive prenylazide groups into peptides using geranylazide diphosphate via protein farnesyltransferase: implications for selective protein labeling. AB - Protein farnesyltransferase (PFTase) catalyzes the attachment of a geranyl azide moiety to a peptide substrate, N-dansyl-Gly-Cys-Val-Ile-Ala-OH. The resulting azide-containing peptide was derivatized with a triphenylphosphine-based reagent to generate an O-alkyl imidate-linked product, rather than the amide-linked material expected via a Staudinger reaction. Since the CAAX box recognition motif (where the internal A residues are aliphatic amino acids) modified by PFTase can be incorporated into the C-terminus of virtually any polypeptide, this two-step procedure provides a general method for incorporating a diverse range of chemical modifications specifically near the C-terminus of proteins. PMID- 15810015 TI - Detection and quantification of circulating tumor cells in mouse models of human breast cancer using immunomagnetic enrichment and multiparameter flow cytometry. AB - BACKGROUND: Circulating tumor cells (CTCs) in the peripheral blood of breast cancer patients may be an important indicator of metastatic disease and poor prognosis. However, the use of experimental models is required to fully elucidate the functional consequences of CTCs. The purpose of this study was to optimize the sensitivity of multiparameter flow cytometry for detection of human tumor cells in mouse models of breast cancer. METHODS: MDA-MB-468 human breast cancer cells were serially diluted in whole mouse blood. Samples were lysed and incubated with a fluorescein isothiocyanate-conjugated anti-human leukocytic antigen antibody and a phycoerythrin-conjugated anti-mouse pan-leukocyte CD45 antibody. Samples were then immunomagnetically depleted of CD45-positive leukocytes, fixed, permeabilized, and stained with propidium iodide before flow cytometric analysis. RESULTS: Human breast cancer cells could be differentiated from mouse leukocytes based on increased light scatter, cell surface marker expression, and aneuploid DNA content. The method was found to have a lower sensitivity limit of 10(-5) and was effective for detecting human breast cancer cells in vivo in the circulation of experimental mice carrying primary human mammary tumors. CONCLUSIONS: This technique has the potential to be a valuable and sensitive tool for investigating the biological relevance of CTCs in experimental mouse models of breast cancer. PMID- 15810016 TI - Directed motility of phagosomes in Tetrahymena thermophila requires actin and Myo1p, a novel unconventional myosin. AB - The phagosome cycle was investigated in Tetrahymena thermophila, which had internalized fluorescent latex beads. Confocal microscopy of cells from a GFP actin strain revealed actin filaments that extended 3-5 mum from the periphery of fluorescent phagosomes. In GFP-actin cells and in wild-type cells, motility of fluorescent phagosomes was directed from the oral cavity to the posterior end of the cell. Although 60% of fluorescent phagosomes in the MYO1-knockout strain were motile, movement of phagosomes was not directed toward the posterior end of the cell and was random. Forty percent of fluorescent phagosomes in knockout cells were non-motile in contrast to only 20% non-motile phagosomes in wild-type cells. The increased incidence of non-motile phagosomes in the knockout strain could reflect absence of Myo1p as a motor. Another myosin or other molecular motors could power random movement of phagosomes in the MYO1-knockout strain. In latrunculin-treated GFP-actin cells, movement of fluorescent phagosomes was random. Average velocity of random movement of fluorescent phagosomes in the knockout strain and in latrunculin-treated cells was statistically the same as the average velocity (2.0 +/- 1.9 microm/min) of phagosomes in GFP-actin cells. These findings are an indication that dynamic actin and Myo1p are required for directed motility of phagosomes. PMID- 15810017 TI - Percutaneous treatment of superior vena cava obstruction following transvenous device implantation. AB - The aim of this study is to assess the feasibility and safety of percutaneous treatment of superior vena cava (SVC) obstruction following transvenous device implantation. SVC obstruction is an uncommon but serious complication that can occur following permanent pacemaker or cardioverter defibrillator implantation utilizing transvenous endocardial leads. The treatment has traditionally been surgical but with the advent of stents, percutaneous approach is becoming popular. We report on the prevalence of SVC obstruction and the safety of its percutaneous catheter-based treatment. This is a retrospective study of SVC obstruction following device implantation in our institution from January 1993 through November 2003. A total of 1,850 permanent pacemaker and 1,200 implantable cardioverter defibrillator initial implants were performed during that period. Three patients developed SVC obstruction following implant (prevalence, 1/1,000 implant). Two patients were males and the mean age at implant was 57 +/- 13 years. Laser lead extraction and SVC angioplasty with or without stenting were performed in all patients. In two of them, this was followed by reimplantation of new systems. There were no procedural complications or mortality. The patients remain free of SVC obstruction symptoms 24 +/- 19 months after treatment. SVC obstruction prevalence after device implantation is low. Percutaneous treatment of SVC obstruction can be safely performed and appears to be effective in maintaining medium-term patency. PMID- 15810018 TI - Simultaneous kissing drug-eluting stent technique for percutaneous treatment of bifurcation lesions in large-size vessels. AB - Treatment of bifurcation lesions is associated with high procedural complications and restenosis rate due to plaque shift, suboptimal angiographic results, difficulty in crossing the stent struts, and incomplete coverage of the side branch ostium. The simultaneous kissing stent (SKS) technique involves two stents, one in main vessel (MV) and one in the side branch (SB) with overlapping stents in the MV proximally, extending proximally the carina of bifurcation. We analyzed our first 200 consecutive patients (202 lesions) who underwent SKS technique for true bifurcation lesions using sirolimus eluting stents, with a minimum follow-up of 6 months. Procedural success was 100% for MV and 99% for SB using SKS technique, with clinical success rate of 97%. In-hospital and 30-day major adverse cardiac events were 3% and 5%, respectively, with a procedure time of 36 +/- 14 min. At mean follow-up of 9 +/- 2 months, the incidence of target lesion revascularization was 4% in the entire group. Therefore, SKS technique using sirolimus-eluting stents may become an effective treatment strategy for large-size bifurcation lesions. However, in order to establish its superiority, SKS technique needs to be compared in a randomized manner with conventional stent techniques. PMID- 15810019 TI - Time to task failure differs with load type when old adults perform a submaximal fatiguing contraction. AB - Young adults exhibit a longer time to task failure when performing a submaximal isometric contraction by pushing against a force transducer (force task) than when supporting an equivalent inertial load (position task). The purpose of this study was to compare the time to failure for old adults when they performed a force task and a position task with the elbow flexor muscles. Eighteen old adults (72 +/- 4 years) performed the force and position tasks at 20% maximal voluntary contraction (MVC) force until task failure. The time to task failure was briefer for the position task (10.6 +/- 6.1 min) than the force task (22.8 +/- 9.1 min, P < 0.05). The rate of increase in electromyographic (EMG) bursting activity, ratings of perceived exertion, mean arterial pressure, heart rate, and fluctuations in motor output during the fatiguing contraction were greater for the position task. However, the increase in averaged EMG for the elbow flexor muscles was greater at termination of the force task. The difference in time to failure for the two tasks was due to a higher level of central neural activity during the position task and was similar to that observed for young adults. These findings indicate that the type of load supported influences the mechanisms and time to task failure for sustained contractions in old adults, and have implications for the design of tasks for rehabilitation and for tasks that minimize fatigue. PMID- 15810020 TI - Association of two mutations in the CHEK2 gene with breast cancer. AB - The 1100delC mutation of the cell cycle checkpoint kinase 2 (CHEK2) gene confers an increased risk for breast cancer, but the clinical impact of other CHEK2 gene variants remains unclear. We determined the frequency of two functionally relevant CHEK2 gene mutations, I157T and IVS2+1G > A, in two large series of breast cancer cases and controls from two independent populations. Our first series consisted of a hospital-based cohort of 996 German breast cancer cases and 486 population controls, and the second series consisted of 424 breast cancer patients and 307 population controls from the Republic of Belarus. The missense substitution I157T was identified in 22/996 cases (2.2%) vs. 3/486 controls (0.6%; OR = 3.6, 95% CI 1.1-12.2, p = 0.044) in the German population and in 24/424 cases (5.7%) vs. 4/307 controls (1.3%; OR = 4.5, 95% CI 1.6-13.2, p = 0.005) in the Byelorussian cohorts. The splicing mutation IVS2+1G > A was infrequent in both populations, being observed in 3/996 German and 4/424 Byelorussian patients (0.3% and 0.9%, respectively) and in 1/486 German controls (0.2%; adjusted OR = 4.0, 95% CI 0.5-30.8, p = 0.273). Heterozygous CHEK2 mutation carriers tended to be diagnosed at an earlier age in both populations, but these differences did not reach statistical significance. Family history of breast cancer did not differ between carriers and noncarriers. Our data indicate that the I157T allele, and possibly the IVS2+1G > A allele, of the CHEK2 gene contribute to inherited breast cancer susceptibility. PMID- 15810021 TI - Polymorphisms in the androgen receptor and the prostate-specific antigen genes and prostate cancer risk. AB - BACKGROUND: Prostate cancer (PCa) is an androgen-dependent disease. Polymorphic CAG and GGC microsatellites in the androgen receptor (AR) can alter transactivation of androgen-responsive genes in in vitro studies. Potentially, this may influence PCa risk. METHODS: Germline DNA samples and survey data were collected from 591 newly diagnosed PCa cases and 538 population-based controls of similar age (40-64 years), from King County, WA. Odds ratios (ORs) and 95% confidence limits were estimated using logistic regression models. RESULTS: No association was detected between PCa and having <22 versus > or =22 CAG repeats (OR = 1.1; 95% CI 0.9, 1.4) or < or =16 GGC versus >16 GGC repeats (OR = 1.0; 95% CI 0.9, 1.4). These findings were unchanged after controlling for body mass index or family history of PCa. No clear relation was detected between APS -158 G/A genotype and risk of PCa or serum prostate-specific antigen (PSA) levels. These findings did not differ by stage or grade of PCa. CONCLUSIONS: We found no evidence that risk of PCa is associated with the AR CAG, GGC, or PSA-158 AREI genetic polymorphisms in middle-aged Caucasian men. PMID- 15810036 TI - Prenatal ultrasonography and neonatal imaging of complete cleft sternum: a case report. AB - The prenatal diagnosis of a complete cleft sternum was established in a fetus of a twin pregnancy at 22 weeks' gestation. We present the prenatal imaging and correlation with postnatal magnetic resonance imaging and high-resolution ultrasonography. Thinned and depressed midline anterior chest wall transmitting the cardiac pulsation was the clue to the diagnosis, and the defective sternum could be identified on close sonographic observation. Successful surgical correction was undertaken at 2 months of age. There were no major associated abnormalities such as ectopia cordis or midline abdominal wall defects. PMID- 15810022 TI - Retrospective evaluation of the dose of Dysport and BOTOX in the management of cervical dystonia and blepharospasm: the REAL DOSE study. AB - The purpose of this study is to evaluate the real-world dose utilization of Dysport and BOTOX for cervical dystonia and blepharospasm. Six investigational sites (five countries) were identified. Investigators abstracted utilization data for patients who received Dysport before switching to BOTOX or BOTOX before switching to Dysport. Patients were identified during scheduled clinic visits and selected if they met study criteria, which included treatment for at least 2 consecutive years (at least 1 year with Dysport or BOTOX, then switched and maintained on BOTOX or Dysport for at least another year). A total of 114 patients were included in the assessment. Ratios of mean dose for Dysport to BOTOX ranged from a low of 2:1 to a high of 11:1. Thirty-one percent of patients fell into the Dysport-to-BOTOX ratio grouping of 5:1 to less than 6:1; 30% of patients had a mean ratio of Dysport to BOTOX of 4:1 to less than 5:1; and only 21% of all patients evaluated fell into the Dysport-to-BOTOX ratio grouping of 3:1 to less than 4:1. Results are consistent with United Kingdom labeling for botulinum toxins stating that units of different serotype A toxins are not interchangeable and simple dose-conversion factors are not applicable. PMID- 15810043 TI - Association of Surgeons of Great Britain and Ireland Annual Meeting. Glasgow, United Kingdom, 13-15 April 2005. Abstracts. PMID- 15810045 TI - Risk-adjusted predictive models of mortality after index arterial operations using a minimal data set. AB - BACKGROUND: Reducing the data required for a national vascular database (NVD) without compromising the statistical basis of comparative audit is an important goal. This work attempted to model outcomes (mortality and morbidity) from a small and simple subset of the NVD data items, specifically urea, sodium, potassium, haemoglobin, white cell count, age and mode of admission. METHODS: Logistic regression models of risk of adverse outcome were built from the 2001 submission to the NVD using all records that contained the complete data required by the models. These models were applied prospectively against the equivalent data from the 2002 submission to the NVD. RESULTS: As had previously been found using the P-POSSUM (Portsmouth POSSUM) approach, although elective abdominal aortic aneurysm (AAA) repair and infrainguinal bypass (IIB) operations could be described by the same model, separate models were required for carotid endarterectomy (CEA) and emergency AAA repair. For CEA there were insufficient adverse events recorded to allow prospective testing of the models. The overall mean predicted risk of death in 530 patients undergoing elective AAA repair or IIB operations was 5.6 per cent, predicting 30 deaths. There were 28 reported deaths (chi(2) = 2.75, 4 d.f., P = 0.600; no evidence of lack of fit). Similarly, accurate predictions were obtained across a range of predicted risks as well as for patients undergoing repair of ruptured AAA and for morbidity. CONCLUSION: A 'data economic' model for risk stratification of national data is feasible. The ability to use a minimal data set may facilitate the process of comparative audit within the NVD. PMID- 15810046 TI - Randomized clinical trial of routine preoperative duplex imaging before varicose vein surgery. AB - BACKGROUND: Duplex imaging is used increasingly for preoperative evaluation of varicose veins, but its value in terms of the long-term results of surgery is not clear. METHODS: Patients with primary varicose veins were randomized to operation with or without preoperative duplex imaging. Reoperation rates, clinical and duplex findings were compared at 2 months and 2 years after surgery. RESULTS: Two hundred and ninety-three patients (343 legs) had varicose vein surgery after duplex imaging (group 1; 166 legs) or no imaging (group 2; 177 legs). In 44 legs (26.5 per cent), duplex examination suggested a different surgical procedure than had been considered on clinical grounds; the procedure was changed accordingly for 29 legs. At 2 months, incompetence was detected at the saphenofemoral or saphenopopliteal junction (or both) in 14 legs (8.8 per cent) in group 1 and in 44 legs (26.5 per cent) in group 2 (P < 0.001). At 2 years, two legs (1.4 per cent) had undergone or were awaiting reoperation in group 1, and 14 legs (9.5 per cent) in group 2 (P = 0.002). In the remainder, major incompetence was found in 19 legs (15.0 per cent) in group 1 and in 53 (41.1 per cent) in group 2 (P < 0.001). CONCLUSION: Routine preoperative duplex examination led to an improvement in results 2 years after surgery for patients with primary varicose veins. PMID- 15810047 TI - Prediction of non-sentinel lymph node status in breast cancer with a micrometastatic sentinel node. AB - BACKGROUND: Axillary lymph node dissection (ALND) may not be necessary in women with breast cancer who have micrometastasis in a sentinel node (SN), owing to the low risk of non-SN (NSN) involvement. The aim of this study was to identify a subgroup of women with a micrometastatic SN and a negligible risk of positive NSNs in whom ALND may be avoided. METHODS: Some 237 of 241 women with a macrometastatic SN and 122 of 138 with a micrometastatic SN underwent completion ALND and were compared with respect to NSN involvement. The 122 patients with SN micrometastasis were further analysed to determine factors that could predict the risk of positive NSNs. RESULTS: A total of 121 (51.1 per cent) of 237 women with SN macrometastasis had positive NSNs compared with 22 (18.0 per cent) of 122 with SN micrometastasis (P < 0.001). Multivariate analysis showed that size of SN micrometastasis (odds ratio 3.49 (95 per cent confidence interval (c.i.) 1.32 to 9.23); P = 0.012) and presence of lymphovascular invasion (odds ratio 0.23 (95 per cent c.i. 0.05 to 1.00); P = 0.050) were significantly associated with positive NSNs. SN micrometastasis less than 0.5 mm in diameter combined with absence of lymphovascular invasion was associated with an 8.5 per cent risk of NSN involvement. CONCLUSION: Size of micrometastasis and presence of lymphovascular invasion were significantly related to the risk of finding additional positive axillary lymph nodes when the SN contained only micrometastasis. PMID- 15810048 TI - Assessing the technical skills of surgical trainees. AB - BACKGROUND: The aims were to determine whether tests of technical skill on simple simulations can predict competence in the operating theatre and whether objective assessment in the operating theatre by direct observation and video recording is feasible and reliable. METHODS: Thirty-three general surgical trainees undertook five simple skill simulations (knotting, skin incision and suturing, tissue dissection, vessel ligation and small bowel anastomosis). The operative competence of each trainee was then assessed during two or three saphenofemoral disconnections (SFDs) by a single surgeon. Video recordings of the operations were also assessed by two surgeons. RESULTS: The inter-rater reliability between direct observation and blinded videotape assessment was high (alpha = 0.96 (95 per cent confidence interval 0.92 to 0.98)). Backward stepwise regression analysis revealed that the best predictors of operative competence were the number of SFDs performed previously plus the simulation scores for dissection and ligation, the key components of SFD (64 per cent of variance explained; P = 0.001). CONCLUSION: Deconstruction of operations into their component parts enables trainees to practise on simple simulations representing each component, and be assessed as competent, before undertaking the actual operation. Assessment of surgical competence by direct observation and video recording is feasible and reliable; such assessments could be used for both formative and summative assessment. PMID- 15810049 TI - Randomized clinical trial of laparoscopic Roux-en-Y gastric bypass versus laparoscopic vertical banded gastroplasty for obesity. AB - BACKGROUND: Laparoscopic techniques have been developed for performing Roux-en-Y gastric bypass (LRYGBP) and vertical banded gastroplasty (LVBG) in patients with morbid obesity. It is not certain, however, which is the better technique in non superobese patients (body mass index less than 50 kg/m(2)). METHODS: Eighty-three patients (LRYGBP 37, LVBG 46) were assessed in a randomized clinical trial. Perioperative complications were recorded together with preoperative and postoperative respiratory function and mobilization rate. Patients were monitored for 2 years after operation with regard to weight change and the need for remedial surgery. RESULTS: There were no conversions to open surgery. The mean operating time was longer for LRYGBP than LVBG (138 versus 105 min). Five early reoperations were performed after LRYGBP (three for haemorrhage, one for ileus and one suspected leak) and one after LVBG (suspected leak). There were no differences in postoperative respiratory function or mobilization. Weight reduction was greater after LRYGBP (excess weight loss 78.3 versus 62.9 per cent 1 year after surgery, P = 0.009; 84.4 versus 59.8 per cent at 2 years, P < 0.001). Remedial surgical intervention was required in eight patients after LVBG (conversion to Roux-en-Ygastric bypass) and none after LRYGBP. CONCLUSION: LRYGBP and LVBG were comparable in terms of operative safety and postoperative recovery, but weight reduction was better after LRYGBP. PMID- 15810050 TI - Effect of supervised surgical training on outcomes after resection of colorectal cancer. AB - BACKGROUND: The process of training surgeons in technique for resection of colorectal cancer should not compromise patient care or outcomes. The aim of this study was to compare morbidity, mortality and survival rates after resection performed by trainees with those for a consultant surgeon. METHODS: Outcomes for 150 patients operated on by a single colorectal surgeon at a private hospital were compared with those of 344 patients admitted under the same surgeon and operated on by closely supervised trainee surgeons in a public teaching hospital between 1995 and 2002. RESULTS: Co-morbidity was significantly more common in patients operated on by trainees; their American Society of Anesthesiologists grades were higher and tumours were more advanced. Of 16 postoperative complications evaluated, only respiratory and cardiac problems were significantly more common in patients operated on by trainees. There was no difference in operative mortality, local recurrence or 2-year survival rate after adjustment for age and tumour stage. CONCLUSION: Outcomes after resection for colorectal cancer did not differ between the consultant and trainees in the context of a closely supervised training programme. PMID- 15810051 TI - Amyloid deposits in transthyretin-derived amyloidosis: cleaved transthyretin is associated with distinct amyloid morphology. AB - The pathological fibrillar deposits found in the heart and other organs of patients with senile systemic amyloidosis (SSA) and Swedish familial amyloidotic polyneuropathy (FAP) contain wild-type (wt) and a mutant form of transthyretin (TTR), respectively. Previously, it was reported that these two forms of amyloid have different molecular features and it was thus postulated that the mechanism responsible for TTR fibrillogenesis in SSA and FAP may differ. To document further the nature of the amyloid in these entities, detailed morphological, histochemical, immunological, and structural analyses of specimens obtained from 14 individuals with SSA and 11 Swedish FAP patients have been performed. Two distinct patterns of amyloid deposition (designated A and B) were evident. In pattern A, found in all SSA and five of 11 FAP cases, the amyloid had a homogeneous but patchy distribution within the sub-endocardium, sub-epicardium, and myocardium; exhibited weak congophilia and green birefringence; and was composed of tightly packed, short, unorientated fibrils. This material contained mainly approximately 79-residue C-terminal fragments of the amyloidogenic precursor protein. In pattern B, seen in the six other FAP patients, the amyloid appeared as thin streaks throughout the cardiac tissue; often surrounded individual muscle cells; was strongly congophilic and birefringent; had long fibrils arranged in parallel bundles, often penetrating into myocytes; and was composed of virtually intact TTR molecules. These findings provide substantive evidence for the morphological and structural heterogeneity of TTR fibrils and suggest that the two types of deposition may reflect fundamental differences in the pathogenesis of the TTR-associated amyloidoses. PMID- 15810055 TI - Critical appraisal of femorofemoral crossover grafts. AB - BACKGROUND: The aim of this study was to determine how often femorofemoral crossover grafting for critical ischaemia or intermittent claudication gives an ideal result. An ideal result is an uncomplicated operation with primary wound healing, relief of ischaemic symptoms without recurrence and no need for further intervention. METHODS: All patients undergoing primary femorofemoral crossover grafting between January 1988 and December 2003 were studied. RESULTS: Some 144 operations were analysed; 51 patients had critical ischaemia and 93 claudication. There was one postoperative death (0.7 per cent). Complications occurred within 30 days in 32 patients (22.2 per cent), including graft occlusion in three (2.1 per cent); six patients (4.2 per cent) required early reoperation. Primary patency for patients with critical ischaemia was 88, 82 and 74 per cent at 1, 3 and 5 years respectively. Respective figures for those who presented with claudication were 93, 92 and 90 per cent (P = 0.034). Late symptoms included graft occlusion (20 patients), disease progression (25), ongoing ulceration (six), graft infection (nine), false aneurysm formation (two) and late donor-site stenosis (two). CONCLUSION: When obtaining informed consent, simply describing patency and limb salvage rates does not provide an accurate picture of the outcome of femorofemoral grafting. PMID- 15810056 TI - Medium-term results of vertical reduction rectoplasty and sigmoid colectomy for idiopathic megarectum. AB - BACKGROUND: Vertical reduction rectoplasty (VRR) was devised specifically to address the physiological abnormalities present in the rectum of patients with idiopathic megarectum (IMR). This study evaluated the medium-term clinical and physiological results of VRR. METHODS: VRR and sigmoid colectomy was performed in ten patients with IMR and constipation (six women). Patients were evaluated before and a median of 60 (range 28-74) months after surgery by assessment of symptoms using scoring systems and anorectal physiological measurements. Independent, detailed postoperative evaluation of rectal diameter, compliance, and sensory and evacuatory function was performed. RESULTS: There were no deaths or late complications. Symptoms recurred necessitating permanent ileostomy formation in two patients. Median (range) constipation scores improved from 22 (18-27) before to 10 (0-24) after surgery (P = 0.016). Median (range) bowel frequency increased from 1.5 (0.2-7) to 7 (0.5-21) per week (P = 0.016). Rectal diameter, compliance and sensory function were normal in seven of eight patients after surgery. Evacuatory function and colonic transit were each normalized in two of eight patients after VRR. CONCLUSION: VRR corrected rectal diameter, compliance and sensory function in most patients, and clinical benefit was sustained in the medium term. The procedure was associated with a low morbidity, and no mortality and should be considered in the surgical management of IMR. PMID- 15810059 TI - Dynamics of Z-band based proteins in developing skeletal muscle cells. AB - During myofibril formation, Z-bodies, small complexes of alpha-actinin and associated proteins, grow in size, fuse and align to produce Z-bands. To determine if there were changes in protein dynamics during the assembly process, Fluorescence Recovery after Photobleaching was used to measure the exchange of Z body and Z-band proteins with cytoplasmic pools in cultures of quail myotubes. Myotubes were transfected with plasmids encoding Yellow, Green, or Cyan Fluorescent Protein linked to the Z-band proteins: actin, alpha-actinin, cypher, FATZ, myotilin, and telethonin. Each Z-band protein showed a characteristic recovery rate and mobility. All except telethonin were localized in both Z-bodies and Z-bands. Proteins that were present both early in development in Z-bodies and later in Z-bands had faster exchange rates in Z-bodies. These results suggest that during myofibrillogenesis, molecular interactions develop between the Z-band proteins that decrease their mobility and increase the stability of the Z-bands. A truncated construct of alpha-actinin, which localized in Z-bands in myotubes and exhibited a very low rate of exchange, led to disruption of myofibrils, suggesting the importance of dynamic, intact alpha-actinin molecules for the formation and maintenance of Z-bands. Our experiments reveal the Z-band to be a much more dynamic structure than its appearance in electron micrographs of cross striated muscle cells might suggest. PMID- 15810067 TI - Portal vein embolization before major hepatectomy. AB - To discuss the rationale, techniques and the unsolved issues regarding preoperative portal vein embolization (PVE) before major hepatectomy. After a systematic search of Pubmed, we reviewed and retrieved literature related to PVE. Preoperative PVE is an approach that is gaining increasing acceptance in the preoperative treatment of selected patients prior to major hepatic resection. Induction of selective hypertrophy of the nondiseased portion of the liver with PVE in patients with either primary or secondary hepatobiliary, malignancy with small estimated future liver remnants (FLR) may result in fewer complications and shorter hospital stays following resection. Additionally, PVE performed in patients initially considered unsuitable for resection due to lack of sufficient remaining normal parenchyma may add to the pool of candidates for surgical treatment. The results suggest that PVE is recommendable in treating the cirrhotic patients before major liver resection. PMID- 15810068 TI - Loss of heterozygosity in multistage carcinogenesis of esophageal carcinoma at high-incidence area in Henan Province, China. AB - AIM: Microsatellites are the repeated DNA sequences scattered widely within the genomes and closely linked with many important genes. This study was designed to characterize the changes of microsatellite DNA loss of heterozygosity (LOH) in esophageal carcinogenesis. METHODS: Allelic deletions in 32 cases of matched precancerous, cancerous and normal tissues were examined by syringe microdissection under an anatomic microscope and microsatellite polymorphism analysis using 15 polymorphic markers on chromosomes 3p, 5q, 6p, 9p, 13q, 17p, 17q and 18q. RESULTS: Microsatellite DNA LOH was observed in precancerous and cancerous tissues, except D9S1752. The rate of LOH increased remarkably with the lesions progressed from basal cell hyperplasia (BCH) to squamous cell carcinoma (SCC) (P<0.05). Three markers, D9S171, D13S260 and TP53, showed the highest incidence of LOH (>60%). LOH loci were different in precancerous and cancerous tissues. LOH in D3S1234 and TP53 was the common event in different lesions from the same patients. CONCLUSION: Microsatellite DNA LOH occurs in early stage of human esophageal carcinogenesis, even in BCH. With the lesion progressed, gene instability increases, the accumulation of this change may be one of the important mechanisms driving precancerous lesions to cancer. PMID- 15810069 TI - Clinical analysis of the risk factors for recurrence of HCC and its relationship with HBV. AB - AIM: To comprehend the risk factors of recurrence of hepatocellular carcinoma (HCC) and its relationship with the infection patterns of hepatitis B virus (HBV). METHODS: All materials of 270 cases of postoperative HCC were statistically analyzed by SPSS software. Recurrence and metastasis were classified into early (< or =2 years) and late phase (>2 years). Risk factors for recurrence and metastasis after surgery in each group were analyzed. RESULTS: Out of 270 cases of HCC, 162 cases were followed up in which recurrence and metastasis occurred in 136 cases. There were a lot of risk factors related to recurrence and metastasis of HCC; risk factors contributing to early phase recurrence were serum AFP level, vascular invasion, incisal margin and operative transfusion, gross tumor classification and number of intrahepatic node to late phase recurrence. The HBV infective rate of recurrent HCC was 94.1%, in which "HBsAg, HBeAb, HbcAb" positive pattern reached 45.6%. The proportion of HBV infection in solitary large hepatocellular carcinoma (SLHCC) evidently decreased compared to nodular hepatocellular carcinoma (NHCC) (P<0.05). CONCLUSION: The early and late recurrence and metastasis after hepatectomy of HCC were associated with different risk factors. The early recurrence may be mediated by vascular invasion and remnant lesion, the late recurrence by tumor's clinical pathology propert, as multicentric carcinogenesis or intrahepatic carcinoma de novo. HBV replication takes a great role in this process. From this study, we found that SLHCC has more satisfactory neoplasm biological behavior than NHCC. PMID- 15810070 TI - Hepatic resection for hepatocellular carcinoma in end-stage renal disease patients: two decades of experience at Chang Gung Memorial Hospital. AB - AIM: Hepatocellular carcinoma (HCC) is a common disease in Taiwan. The prevalence of viral hepatitis infection and the subsequent development of HCC are well known to be higher in patients with end-stage renal disease (ESRD) requiring hemodialysis (HD) or peritoneal dialysis (PD) than among the general population. However, information on hepatic resection for ESRD-HCC patients is limited. METHODS: The clinical features of 26 ESRD-HCC patients who underwent hepatic resection from 1982 to 2001 were retrospectively reviewed. Meanwhile, the clinicopathological features and the outcome of 1 198 HCC patients without ESRD undergoing hepatic resection were used for comparison. RESULTS: Of 1 224 surgically resected HCC patients, 26 (4.2%) were ESRD-HCC. Univariate analysis revealed more associated disease, more physical signs of anemia and postoperative complications, lower hemoglobin, platelet, alpha-fetoprotein, elevated blood urea nitrogen (BUN) and creatinine levels, smaller tumors, lower HBsAg positivity, higher HCV positivity, and longer hospital stays in the ESRD-HCC group compared with the HCC group. Furthermore, multivariate stepwise logistic regression analysis revealed that elevated BUN and creatinine levels were the only two independently significant factors in the patients in the ESRD-HCC group. Overall and disease-free survival rates were similar between the ESRD-HCC and HCC groups. CONCLUSION: Elevated BUN and creatinine were the only two main independent factors differentiating ESRD-HCC from HCC patients. ESRD should not be a contraindication of hepatic resection in HCC patients; however, careful operative techniques and perioperative care are crucial to achieving lower morbidity and mortality. Comparable overall survival and disease-free survival can be achieved in selected ESRD-HCC patients undergoing hepatic resection when compared with conventional HCC patients. PMID- 15810071 TI - Expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90, GRP78, GRP94) in hepatitis B virus-related hepatocellular carcinomas and dysplastic nodules. AB - AIM: Expression of heat shock proteins (HSPs) is frequently up-regulated in hepatocellular carcinoma (HCC), which evolves from dysplastic nodule (DN) and early HCC to advanced HCC. However, little is known about the differential expression of HSPs in multistep hepatocarcinogenesis. It was the purpose of this study to monitor the expression of HSPs in multistep hepatocarcinogenesis and to evaluate their prognostic significance in hepatitis B virus (HBV)-related HCC. METHODS: Thirty-eight HCC and 19 DN samples were obtained from 52 hepatitis B surface antigen-positive Korean patients. Immunohistochemical and dot immunoblot analyses of HSP27, HSP60, HSP70, HSP90, glucose regulated protein (GRP)78, and GRP94 were performed and their expression at different stages of HCC development was statistically analyzed. RESULTS: Expression of HSP27, HSP70, HSP90, GRP78, and GRP94 increased along with the stepwise progression of hepatocarcinogenesis. Strong correlation was found only in GRP78 (Spearman's r = 0.802). There was a positive correlation between the expressions of GRP78, GRP94, HSP90, or HSP70 and prognostic factors of HCC. Specifically, the expression of GRP78, GRP94, or HSP90 was associated significantly with vascular invasion and intrahepatic metastasis. CONCLUSION: The expressions of HSPs are commonly up-regulated in HBV-related HCCs and GRP78 might play an important role in the stepwise progression of HBV-related hepatocarcinogenesis. GRP78, GRP94, and HSP90 may be important prognostic markers of HBV-related HCC, strongly suggesting vascular invasion and intrahepatic metastasis. PMID- 15810072 TI - Controlled and reversible induction of differentiation and activation of adult human hepatocytes by a biphasic culture technique. AB - AIM: Clinical application of human hepatocytes (HC) is hampered by the progressive loss of growth and differentiation in vitro. The object of the study was to evaluate the effect of a biphasic culture technique on expression and activation of growth factor receptors and differentiation of human adult HC. METHODS: Isolated HC were sequentially cultured in a hormone enriched differentiation medium (DM) containing nicotinamide, insulin, transferrin, selenium, and dexame-thasone or activation medium (AM) containing hepatocyte growth factor (HGF), epidermal growth factor (EGF), and granulocyte-macrophage colony-stimulating factor (GM-CSF). Expression, distribution and activation of the HC receptors (MET and EGFR) and the pattern of characteristic cytokeratin (CK) filaments were measured by fluorometry, confocal microscopy and Western blotting. RESULTS: In the biphasic culture system, HC underwent repeated cycles of activation (characterized by expression and activation of growth factor receptors) and re-differentiation (illustrated by distribution of typical filaments CK-18 but low or absent expression of CK-19). In AM increased expression of MET and EGFR was associated with receptor translocation into the cytoplasm and induction of atypical CK-19. In DM low expression of MET and EGFR was localized on the cell membrane and CK-19 was reduced. Receptor phosphorylation required embedding of HC in collagen type I gel. CONCLUSION: Control and reversible modulation of growth factor receptor activation of mature human HC can be accomplished in vitro, when defined signals from the extracellular matrix and sequential growth stimuli are provided. The biphasic technique helps overcome de-differentiation, which occurs during continuous stimulation by means of growth factors. PMID- 15810073 TI - Identification of the immunogenic domains in HBsAg preS1 region using overlapping preS1 fragment fusion proteins. AB - AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitope-based hepatitis B vaccine design. METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA. RESULTS: The results in mice showed that the immu-nogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity. More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing. CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic dom-ains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34 59) fragments into epitope-based HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans. PMID- 15810074 TI - Effects of emodin and baicalein on rats with severe acute pancreatitis. AB - AIM: To investigate the therapeutic effects of emodin in combination with baicalein on severe acute pancreatitis (SAP) rats and to explore the mechanism of SAP. METHODS: A total of 112 SAP rats induced by retrograde injection of 5% sodium taurocholate into the biliary-pancreatic duct, randomly assigned to a untreated group and three treated groups emodin group, combined emodin and baicalein group, and sandostatin group. Meanwhile, another 28 other rats were selected as sham operation (SO) group. There were 28 rats in each group, 8 rats were in 3 and 6 h groups respectively, and 12 rats in 12 h group. At each time points, survival rates, ascites volumes, pathological lesion scores of pancreas tissues, serum amylase, tumor necrosis factor-alpha and IL-6 levels were determined as the indexes of therapeutic effects. RESULTS: The survival rate at 12 h was significantly higher in three treated groups than in untreated group. The ascites volume at 12 h was remarkably less in combined and sandostatin groups than in emodin group, but there was no difference between combined group and sandostatin group (P>0.05). Serum amylase levels at all time-points were significantly lower in three treated groups than in untreated group. However, they had no difference among treated groups (P>0.05). Serum TNF-alpha were lower in three treated groups than in untreated group at all time points. Among the three treated groups, at 6 h, the TNF-alpha levels of combination and sandostatin groups were lower than those of emodin group. These was no difference between combined and sandostantin. Serum IL-6 concentration at 3 h were lower in combined and sandostatin groups than in untreated group, but at 6 and 12 h they were lower in all treated groups than in untreated group and the combined and sandostatin groups and in emodin group, no difference was found between combined and sandostatin groups at all time-points (P>0.05). The pathological scores of pancreas at all time points were significantly lower in three treated groups than in the untreated group, and at 6, 12 h, the scores of combined and sandostatin groups were lower than in emodin group. There was no difference between combined and sandostatin groups (P>0.05). CONCLUSION: Combination of emodin with baicalein has significant therapeutic effects on SAP rats. PMID- 15810075 TI - Chromic-P32 phosphate treatment of implanted pancreatic carcinoma: mechanism involved. AB - AIM: To study the effects of chromic-P32 phosphate (32P colloids) interstitial administration in Pc-3 implanted pancreatic carcinoma, and investigate its anticancer mechanism. METHODS: Ninety-eight tumor bearing nude mice were killed at different time points after the injection of 32P colloids to the tumor core with observed radioactivity. The light microscopy, transmission electron microscopy (TEM) and immuno-histochemistry and flow cytometry were used to study the rates of tumor cell necrosis, proliferating cell nuclear antigen index, the micro vessel density (MVD). The changes of the biological response to the lymphatic transported 32P colloids in the inguinal lymph node (ILN) were dynamically observed, and the percentage of tumor cell apoptosis, and Apo2.7, caspase-3, Bcl-2, Bax-related gene expression were observed too. RESULTS: The half-life of effective medication is 13 d after injection of 32P colloids to the tumor stroma, in 1-6 groups, the tumor cell necrosis rates were 20%, 45%, 65%, 70%, 95% and 4%, respectively (F = 4.14-105.36, P<0.01). MVD were 38.5+/-4.0, 28.0+/-2.9, 17.0+/-2.9, 8.8+/-1.5, 5.7+/-2.3 and 65.0+/-5.2 (t = 11.9-26.1, P<0.01), respectively. Under TEM fairly differentiated Pc-3 cells were found. Thirty days after medication, tumors were shrunk and dried with scabs detached, and those in control group increased in size prominently with plenty of hypodermic blood vessels. In all animals the ILN were enlarged but in medicated animals they appeared later and smaller than those in control group. The extent of irradiative injury in ILN was positively correlated to the dosage of medication. Typical tumor cell apoptosis could be found under TEM in animals with intra-tumoral injection of low dosed 32P colloids. The peak of apoptosis occurred in 2.96 MBq group and 24 h after irradiation. In the course of irradiation induced apoptosis, the value of Bcl-2/Bax was down regulated; Apo2.7 and caspase 3 protein expression were prominently increased dose dependently. CONCLUSION: 32P colloids intra-tumor injection having prominent anticancer effectiveness may reveal the ability of promoting cell differentiation. The low dose 32P colloids may induce human pancreatic carcinoma Pc-3 implanted tumor cell apoptosis; Apo2.7, caspase-3, Bcl-2 and Bax protein participated in regulating the process of irradiation induced cell apoptosis. PMID- 15810076 TI - Expression profiling suggests a regulatory role of gallbladder in lipid homeostasis. AB - AIM: To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis. METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized to a cDNA array representing 17,000 cDNA clusters. Genes with intensities > or =2 and variation <0.33 between two samples were considered as positive signals with subtraction of background chosen from an area where no cDNA was spotted. The average gray level of two gallbladders was adopted to analyze its bioinformatics. Identified target genes were confirmed by touch-down polymerase chain reaction and sequencing. RESULTS: A total of 11 047 genes expressed in normal gallbladder, which was more than that predicted by another author, and the first 10 genes highly expressed (high gray level in hybridization image), e.g. ARPC5 (2 225.88+/-90.46), LOC55972 (2 220.32+/-446.51) and SLC20A2 (1 865.21+/-98.02), were related to the function of smooth muscle contraction and material transport. Meanwhile, 149 lipid-related genes were expressed in the gallbladder, 89 of which were first identified (with gray level in hybridization image), e.g. FASN (11.42+/-2.62), APOD (92.61+/-8.90) and CYP21A2 (246.11+/-42.36), and they were involved in each step of lipid metabolism pathway. In addition, 19 of those 149 genes were gallstone candidate susceptibility genes (with gray level in hybridization image), e.g. HMGCR (10.98+/-0.31), NPC1 (34.88+/-12.12) and NR1H4 (16.8+/-0.65), which were previously thought to be expressed in the liver and/or intestine tissue only. CONCLUSION: Gallbladder expresses 11 047 genes and takes part in lipid homeostasis. PMID- 15810077 TI - Beta-catenin up-regulates the expression of cyclinD1, c-myc and MMP-7 in human pancreatic cancer: relationships with carcinogenesis and metastasis. AB - AIM: To investigate whether abnormal expression of beta-catenin in conjunction with overexpression of cyclinD1, c-myc and matrix metalloproteinase-7 (MMP-7) correlated with the carcinogenesis, metastasis and prognosis of pancreatic cancer, and to analyze the relationship of beta-catenin expression with cyclinD1, c-myc and MMP-7 expression. METHODS: Using immunohistochemistry, we examined the expression of beta-catenin, cyclinD1, c-myc and MMP-7 in 47 pancreatic adenocarcinoma tissues, 12 pancreatic intraepithelial neoplasia (PanIN) and 10 normal pancreases, respectively. Proliferation cell nuclear antigen was also tested as the index of proliferative activity of pancreatic cancer cells. RESULTS: In 10 cases of normal pancreatic tissues, epithelial cells showed equally strong membranous expression of beta-catenin protein at the cell-cell boundaries, but the expression of cyclinD1, c-myc and MMP-7 was negative. The expression of beta-catenin, cyclinD1, c-myc and MMP-7 in PanIN and pancreatic adenocarcinoma tissues had no significant difference [6/12 and 32/47 (68.1%), 6/12 and 35/47 (74.5%), 5/12 and 33/47 (70.2%), 7/12 and 30/47 (63.8%), respectively]. The abnormal expression of beta-catenin was significantly correlated to metastasis and one-year survival rate of pancreatic cancer, but had no relation with size, differentiation and cell proliferation. The expression of cyclinD1 was correlated with cell proliferation and extent of differentiation, but not with size, metastasis and one-year survival rate of the pancreatic cancer. The expression of c-myc was not correlated with size, extent of differentiation, metastasis and 1-year survival rate, but closely with cell proliferation of pancreatic cancer. The overexpression of MMP-7 was significantly associated with metastasis and 1-year survival rate of pancreatic cancer, but not with size, extent of differentiation and cell proliferation. There was a highly significant positive association between abnormal expression of beta-catenin and overexpression of cyclinD1, c-myc and MMP-7 not only in PanIN (r = 1.000, 0.845, 0.845), but also in pancreatic cancer (r = 0.437, 0.452, 0.435). CONCLUSION: The abnormal expression of beta-catenin plays a key role in the carcinogenesis and progression of human pancreatic carcinoma by up-regulating the expression of cyclinD1, c-myc and MMP-7, resulting in the degradation of extracellular matrix and uncontrolled cell proliferation and differentiation. beta-catenin abnormal expression and MMP-7 overexpression may be considered as two useful markers for determining metastasis and prognosis of human pancreatic cancer. PMID- 15810078 TI - Effects of total glucosides of peony on immunological hepatic fibrosis in rats. AB - AIM: To study the effects of total glucosides of peony (TGP) on immunological hepatic fibrosis induced by human albumin in rats. METHODS: Sixty adult male Sprague-Dawley rats were randomly divided into: Normal group, model group, TGP (60 and 120 mg/kg) treatment groups and colchicines (0.1 mg/kg) treatment group. On the day before the rats were killed, those in TGP or colchicine groups received TGP or colchicine as above from the first day of tail vein injection of human albumin. The rats in normal and model groups were only administered with the same volume of vehicle. At the end of the 16th wk, rats in each group were killed. Blood and tissue specimens were taken. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), content of malondialdehyde (MDA), activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px), were measured by biochemical methods. Serum procollagen type III (PC III) and laminin (LN) were determined by radioimmunoassay. Liver collagen level was determined by measuring hydroxyproline content in fresh liver samples. Hepatic tissue sections were stained with hematoxylin-eosin and examined under a light microscope. RESULTS: Histological results showed that TGP improved the human albumin-induced alterations in the liver structure, alleviated lobular necrosis and significantly lowered collagen content. The antifibrotic effect of TGP was also confirmed by decreased serum content of LN and PCIII in TGP-treated group. Moreover, the treatment with TGP effectively reduced the hydroxyproline content in liver homogenates. However, the level of ALT and AST increased in fibrotic rat but had no significance compared with normal control, whereas the ratio of A/G decreased without significance. TGP had no effect on level of ALT, AST and the ratio of A/G. Furthermore, TGP treatment significantly blocked the increase in MDA and NO, associated with a partial elevation in liver total antioxidant capacity including SOD and GSH-px. CONCLUSION: TGP has beneficial effects on hepatic fibrosis in rats by inhibition of collagen synthesis and decreasing oxidative stress. PMID- 15810079 TI - Effects of STI571 and p27 gene clone on proliferation and apoptosis of K562 cells. AB - AIM: To investigate the combined effect of STI571 and p27 gene clone on the regulation of proliferation, cell cycle and apoptosis of K562 cell line. METHODS: p27 gene was obtained by RT-PCR, and its sequence was approved to be correct. Then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line. p27 pcDNA3.1-K562 cell clone was screened by G418 after transfection, p27 protein was identified by Western blot. MTT was used to detect the survival rate of the cell. Flow cytometry was used to detect cell cycle and apoptosis index. RESULTS: The expression of p27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. A strong inhibition of cell proliferation was observed in p27-pcDNA3.1 K562 cells as compared with that of the control (pcDNA3.1-K562 cells). The cells at G0/G1 phase were significantly increased, and cells at S phase were greatly declined. The apoptosis index was increased greatly after p27-pcDNA3.1-K562 cells were treated with STI571, and survival rate of the cell was markedly declined (0.35-0.58, P<0.05-0.048 vs STI571-K562 cell, 0.35-0.72, P<0.01-0.001 vs p27-K562 cell). CONCLUSION: p27 and STI571 have a synergistic action on inhibition of proliferation and induction of apoptosis on K562 cells. PMID- 15810080 TI - Hepatic preconditioning of doxorubicin in stop-flow chemotherapy: NF kappaB/IkappaB-alpha pathway and expression of HSP72. AB - AIM: To provide hepatic protection through administration of doxorubicin before stop-flow chemotherapy (SFC) and to investigate the expression of heat shock protein 72 (HSP72) and role of nuclear factor kappa B (NF-kappaB) in this effect. METHODS: The hepatic preconditioning of doxorubicin was established in a porcine model by injection of doxorubicin (1 mg/kg) before SFC. The experimental animals were randomized into two groups: groups receiving doxorubicin (DOX) and normal saline (NS). Serial serum and tissue samples were taken from both groups to evaluate the protection of doxorubicin. Western blot and immuno-precipitation were applied to detect the expression of HSP72, NF-kappaB p65 protein, inhibitor kappaB-alpha (IkappaB-alpha) and phosphorylated IkappaB-alpha as well. The expression of tumor necrosis factor alpha (TNF-alpha) was estimated by semiquantitative RT-PCR. And the extent of the hepatic injury was estimated with the level of serum aminotransferases. RESULTS: An abundance production of HSP72 in porcine liver was observed after 24 h of intravenous administration of doxorubicin, but without any change in the expression of NF-kappaB p65 subunit in cytoplasm. NF-kappaB p65 subunit accumulated in nuclei at the end of SFC and reached its highest level at 30 min after the restoration of the abdominal circulation and decreased gradually during the 6 h after SFC in NS group, while there was little change in DOX group. There was also a slight decrease of IkappaB alpha at 30 min after the restoration of the abdominal circulation in NS group accompanying with the appearance of phosphorylated IkappaB-alpha. The expression of TNF-alpha was significantly higher in NS group than that in DOX group (average 1.40+/-0.17 vs 0.62+/-0.22, P<0.01) at serial time points after SFC. Serum ALT and AST levels of NS group were higher after 24 h than those of DOX group (93.2+/ 7.8 IU/L vs 53.3+/-13.9 IU/L, 217.0+/-29.4 IU/L vs 155.0+/-15.6 IU/L for ALT and AST respectively, P<0.05) and after 48 h than those of DOX group (66.6+/-18.1 IU/L vs 43.3+/-16.7 IU/L, 174.4+/-21.3 IU/L vs 125.7+/-10.5 IU/L for ALT and AST respectively, P<0.05). CONCLUSION: Doxorubicin renders the liver to be tolerant to the hepatic influence in SFC in a porcine model through the NF-kappaB/IkappaB alpha pathway with the expression of HSP72. PMID- 15810081 TI - Value of CT in the diagnosis and management of gallstone ileus. AB - AIM: To retrospectively establish the diagnostic criteria of gallstone ileus on CT, and to prospectively apply these criteria to determine the diagnostic accuracy of CT to confirm or exclude gallstone ileus in patients who presented with acute small bowel obstruction (SBO). Another purpose was to ascertain whether the size of ectopic gallstones would affect treatment strategy. METHODS: Fourteen CT scans in cases of proved gallstone ileus were evaluated retrospectively by two radiologists for the presence or absence of previously reported CT findings to establish the diagnostic criteria. These criteria were applied in a prospective contrast enhanced CT study of 165 patients with acute SBO, which included those 14 cases of gallstone ileus. The hard copy images of 165 CT studies were reviewed by a different group of two radiologists but without previous knowledge of the patient's final diagnosis. All CT data were further analyzed to determine the diagnostic accuracy of gallstone ileus when using CT in prospective evaluation of acute SBO. The size of ectopic gallstone on CT was correlated with the clinical course. RESULTS: The diagnostic criteria of gallstone ileus on CT were established retrospectively, which included: (1) SBO; (2) ectopic gallstone; either rim-calcified or total-calcified; (3) abnormal gall bladder with complete air collection, presence of air-fluid level, or fluid accumulation with irregular wall. Prospectively, CT confirmed the diagnosis in 13 cases of gallstone ileus with these three criteria. Only one false negative case could be identified. The remaining 151 patients are true negative cases and no false positive case could be disclosed. The overall sensitivity, specificity and accuracy of CT in diagnosing gallstone ileus were 93%, 100%; and 99%, respectively. Surgical exploration was performed in 13 patients of gallstone ileus with ectopic stones sized larger than 3 cm. One patient recovered uneventfully following conservative treatment with an ectopic stone sized 2 cm in the long axis. CONCLUSION: Contrast enhanced CT imaging offered crucial evidence not only for the diagnosis of gallstone ileus but also for decision making in management strategy. PMID- 15810082 TI - Clinical significance of hepatic derangement in severe acute respiratory syndrome. AB - AIM: Elevation of alanine aminotransferase (ALT) level is commonly seen among patients suffering from severe acute respiratory syndrome (SARS). We report the progression and clinical significance of liver derangement in a large cohort of SARS patient. METHODS: Serial assay of serum ALT was followed in patients who fulfilled the WHO criteria of SARS. Those with elevated ALT were compared with those with normal liver functions for clinical outcome. Serology for hepatitis B virus (HBV) infection was checked. Adverse outcomes were defined as oxygen desaturation, need of intensive care unit (ICU) and mechanical ventilation and death. RESULTS: Two hundred and ninety-four patients were included in this study. Seventy (24%) patients had elevated serum ALT on admission and 204 (69%) patients had elevated ALT during the subsequent course of illness. Using peak ALT >5XULN as a cut-off and after adjusting for potential confounding factors, the odds ratio of peak ALT >5X ULN for oxygen desaturation was 3.24 (95%CI 1.23-8.59, P = 0.018), ICU care was 3.70 (95%CI 1.38-9.89, P = 0.009), mechanical ventilation was 6.64 (95%CI 2.22-19.81, P = 0.001) and death was 7.34 (95%CI 2.28-24.89, P = 0.001). Ninety-three percent of the survived patients had ALT levels normalized or were on the improving trend during follow-up. Chronic hepatitis B was not associated with worse clinical outcomes. CONCLUSION: Reactive hepatitis is a common complication of SARS-coronavirus infection. Those patients with severe hepatitis had worse clinical outcome. PMID- 15810083 TI - Systemic immune responses to oral administration of recombinant attenuated Salmonella typhimurium expressing Helicobacter pylori urease in mice. AB - AIM: To evaluate whether attenuated Salmonella typhimurium producing Helicobacter pylori (H pylori) urease subunit B (UreB) could induce systemic immune responses against H pylori infection. METHODS: Attenuated S. typhimurium SL3261 was used as a live carrier of plasmid pTC01-UreB, which encodes recombinant H pylori UreB protein. Balb/c mice were given oral immunization with two doses of SL3261/pTC01 UreB at a 3-wk interval. Twelve weeks after oral immunization of mice, serum IgG antibodies were evaluated by ELISA assay. Gamma interferon (IFN-gamma) and interleukin 10 (IL-10) in the supernatant of spleen cell culture were also assessed by ELISA. RESULTS: After oral immunization of mice, serum specific IgG antibodies against UreB in vaccine group were much higher than that in PBS and native Salmonella SL3261 control groups (A450, 0.373+/-0.100 vs 0.053+/-0.022, 0.142+/-0.039, respectively, P<0.01). Moreover, IFN-gamma in vaccine group was on average 167.53+/-29.93 pg/mL, which showed a significant increase vs that of PBS control group (35.68+/-3.55 pg/mL, P<0.01). There was also a tremendous increase of IL-10 in vaccine group compared to PBS and SL3261 control groups (275.13+/ 27.65 pg/mL vs 56.00+/-7.15 pg/mL, 68.02+/-15.03 pg/mL, respectively, P<0.01). In addition, no obvious side effects in mice and no change in gastric inflammation were observed. CONCLUSION: The multiple oral immunizations with the attenuated S. typhimurium expressing H pylori UreB could induce significant systemic immune responses, suggesting it may be used as oral vaccine against H pylori infection. PMID- 15810084 TI - Effect of hepatitis C virus nonstructural protein NS3 on proliferation and MAPK phosphorylation of normal hepatocyte line. AB - AIM: To study the effect of hepatitis C virus nonstructural region 3 (HCV NS3) protein on proliferation and transformation of normal human liver cell line. METHODS: QSG7701 cells were transfected with pRcHCNS3-5', pRcHCNS3-3' and pRcCMV using lipofectamine transfecting technique and selected with G418 method. Expression of HCV NS3 protein was determined by immunohistochemistry. Biologic characteristics of transfected cells were evaluated by population doubling time and soft agar assays. Activation of MAPK was analyzed using Western blot with phosphospecific monoclonal antibody against dually phosphorylated MAPK. RESULTS: QSG7701 cells transfected with pRcHCNS3-5' showed strong intracellular expression of HCVNS3 protein, and the positive signal was localized in cytoplasm. The expressing strength of HCVNS3 protein in pRcHCNS3-3'-transfected cells was weaker than that in pRcHCNS3-5'-transfected cells. The population doubling time in the transfected cells with pRcHCNS3-5' (12 h) was much shorter than those with pRcHCNS3-3', pRcCMV and normal cells (24, 26, 28 h, respectively) (P<0.01). The transfected cells with pRcHCNS3-5' showed much more anchorage independent colonies than that in those with pRcHCNS3-3' and pRcCMV (P<0.01). The cloning efficiencies of transfected cells with pRcHCNS3-5', pRcHCNS3-3', pRcCMV and controls were 33%, 1.33%, 1.46%, 1.11% respectively. The level of phosphorylated MAPK in the cells with pRcHCNS3-5' was much higher than that in those with pRcHCNS3-3'and pRcCMV and normal cells (P<0.01). CONCLUSION: The results suggest that (1) QSG7701 cells are a better human liver cell line for investigating the pathogenesis of HCV NS3 protein. (2) 5' region of the HCV genome segment encoding HCV NS3 is involved in cell growth and cell phenotype. (3) HCV NS3 N-terminal peptide may up-regulate the activation of MAPK, but not affect the expression of MAPK. PMID- 15810085 TI - Clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer. AB - AIM: To study the clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer. METHODS: To investigate the expression of p53 and mdm2 in pancreatic cancer by immunohistochemistry, and the relationships between the p53 and mdm2 protein expression and clinicopathological parameters in pancreatic cancer. RESULTS: The positive expression of p53 protein was found in 40 of 59 patients (67.8%) and that of mdm2 protein in 17 of 59 patients (28.8%). No obvious relationships were found between p53 as well as mdm2 expression and sex, tumor site, TNM staging and histological differentiation. p53 expression was increased in patients younger than 65 years old, while mdm2 had no relationship with age. The survival time of the patients with the positive expression of p53 and mdm2 proteins was obviously shorter than the other groups. CONCLUSION: Both p53 and mdm2 presented relatively high expression in human pancreatic cancer. The overexpression of p53 and mdm2 might reflect the malignant proliferation of pancreatic cancer and their co-expression might be helpful to evaluate the prognosis of the patients with pancreatic cancer. PMID- 15810086 TI - Plasma von Willebrand factor level as a prognostic indicator of patients with metastatic colorectal carcinoma. AB - AIM: To evaluate the correlations of plasma von Willebrand factor (vWF) level with the distant metastasis and prognosis of patients with colorectal cancer. METHODS: A total of 86 patients with histologically confirmed metastatic colorectal cancers receiving treatment at Taipei Veterans General Hospital were enrolled. All patients had measurable metastatic lesions and life expectancies of more than 3 mo. Plasma vWF levels were measured by immuno-turbidimetric assay and compared with results from 40 non-metastatic colorectal cancer patients and 22 healthy controls. Patients with metastatic colorectal cancer were divided into two groups according to serum vWF levels and the differences between these two groups were analyzed using chi2 test. Data on age, gender, performance status, location of primary tumor, extent of metastasis, site of metastases, histological differentiation, serum CEA and plasma vWF levels were analyzed to determine association with survival. Survival curves were constructed by Kaplan-Meier product limit method and the data was analyzed using log-rank test on a microcomputer. Multivariate analysis using the Cox's proportional hazards regression model was then performed to determine the independent prognostic indicators among all of the possible variables. RESULTS: Colorectal cancer patients were identified as having significantly higher plasma vWF concentrations than healthy controls (P<0.05). Moreover, higher vWF plasma levels were associated with advanced tumor stage (P<0.05) and the presence of multiple metastases (P = 0.014). Patients with lower vWF plasma levels (< or =160%) survived significantly longer than those with a higher plasma vWF level (log-rank test, P = 0.0043). By multivariate analysis, plasma vWF levels (P<0.001), the extent of metastasis (P = 0.012), and the performance status (P = 0.014) were identified as independent prognostic factors. CONCLUSION: Our data indicates that high plasma vWF concentrations correlate with advanced diseases and significantly poor prognosis of patients with metastatic colorectal carcinoma. It may serve as a potential biological marker of disease progression in these patients. PMID- 15810087 TI - Outcome of gall bladder polypoidal lesions detected by transabdominal ultrasound scanning: a nine year experience. AB - AIM: To determine the outcome of polypoidal lesions within the gall bladder (PLG) diagnosed by trans-abdominal scanning. METHODS: A nine-year (1993-2002) retrospective case-note review of all patients who underwent ultrasound scanning after referral to a single Upper GI Surgeon at a District General Hospital was conducted. Patients who were diagnosed with a PLG were included in our study. A database was constructed and patient details, investigations including ultrasound scan (USS) findings, treatment and histology and final diagnosis were recorded. RESULTS: Twenty-three (out of 651) patients were diagnosed pre-operatively by USS to have a polyp-like gall bladder lesion (PLG). Post cholecystectomy histological examination revealed 12 gallstones, 7 cholesterol polyps, 3 adenocarcinomas within polyps and 1 normal gall bladder. The specificity of USS in the diagnosis of PLG was 92.3%. All the true polyps were malignant. Overall USS had 66.66% sensitivity and 100% specificity in the pre-operative suspicion of malignancy. Using size greater than 10 mm as measured on USS as a cut-off, we find 100% sensitivity and 86.95% specificity with a positive predictive value of 50% in the diagnosis of malignancy in PLG. CONCLUSION: A large number of PLG are in fact calculi within diseased gall bladder. In cases of gall bladder polyps more then 10 mm in size on USS further imaging (cross-sectional and/or EUS) is indicated prior to surgery. This will help in the optimal management of patients and avoid histological surprises. PMID- 15810088 TI - Risk factors for the recurrence of hepatocellular carcinoma after radiofrequency ablation of hepatocellular carcinoma in patients with hepatitis C. AB - AIM: To analyze the risk factors of hepatocellular carcinoma (HCC) recurrence after radiofrequency ablation (RFA) treatment with HCV-associated hepatitis. METHODS: Twenty-six patients with HCV-associated HCC who were followed-up for more than 12 mo were selected for this study. Risk factors for distant intrahepatic recurrences of HCC were evaluated for patients in whom complete coagulation was achieved without recurrence in the same subsegment as the primary nodule. Twelve clinical and tumoral factors were examined: Age, gender, nodule diameter, number of primary HCC nodule, Child-Pugh classification, serum platelet, serum albumin, serum AST, post RFA AST, serum ALT, post RFA ALT, post RFA treatment. RESULTS: Distant recurrences of HCC in remnant liver after RFA were observed in 14 cases and in the number of primary HCC nodules (P = 0.047), and the serum platelets (P = 0.030), the clear difference came out by the recurrence group and the non-recurrence group. The cumulative recurrence rates after 1 and 2 years were 30.8% and 86.8%, respectively for primary multinodular HCC, and 15.4% and 29.5% respectively, for primary uninodular HCC. In addition the 1-year recurrence rates for patients with serum albumin more than 3.4 g/dL and less than 3.4 g/dL were 23.1% for both, but the 2-years recurrence rates were 89.0% and 23.1%, respectively. The number of primary HCC nodules (relative risk, 6.970; P = 0.016) were found to be a statistically significant predictor for poor distant intrahepatic recurrence by univariate analysis. CONCLUSION: Patients who have multiple HCC nodules, low serum platelets and low serum albumin accompanied by HCV infection, should be carefully followed because of the high incidence of new HCC lesions in the remnant liver, even if coagulation RFA is complete. PMID- 15810089 TI - Relationships of tumor inflammatory infiltration and necrosis with microsatellite instability in colorectal cancers. AB - AIM: The relationships between microsatellite instability (MSI) and survival in colorectal cancer patients are not consistent. The favorable survival of patient with MSI has been suggested to be related to pronounced inflammatory infiltration; however, the reason for non-association of MSI with survival is unclear. Our aims were to investigate the associations of inflammatory infiltration and tumor necrosis (TN) with microsatellite status and clinicopathological factors in colorectal cancer patients in whom MSI was not related to survival. METHODS: Three hundred and one colorectal adenocarcinomas were evaluated for inflammatory infiltration and 300 for TN under light microscope. RESULTS: Low infiltration at invasive margin (chi2 = 3.94, P = 0.047) and in whole tumor stroma (chi2 = 3.89, P = 0.049) was associated with MSI, but TN was not (chi2=0.10, P = 0.75). Low infiltration was related to advanced stage (chi2 = 8.67, P = 0.03), poorer differentiation (chi2 = 8.84, P = 0.03), DNA non diploid (chi2 = 10.04, P = 0.002), higher S-phase fraction (chi2 = 11.30, P = 0.004), positive p53 expression (chi2 = 7.94, P = 0.01), and worse survival (P = 0.03 for both univariate and multivariate analyses). Abundant TN was related to advanced stage (chi2 = 17.74, P = 0.001) and worse survival (P = 0.02 for univariate, and P = 0.05 for multivariate analysis). CONCLUSION: The result that high inflammatory infiltration was not related to MSI might help explain the non association of MSI with survival in colorectal cancer patients. PMID- 15810090 TI - Changes of ECM and CAM gene expression profile in the cirrhotic liver after HCV infection: analysis by cDNA expression array. AB - AIM: We aimed to observe the expression of extracellular matrix (ECM) and cellular adhesion molecules (CAM) in cirrhotic liver tissues after hepatitis C virus (HCV) infection. METHODS: Twelve patients with post HCV inflammatory liver cirrhosis were selected to evaluate their liver function and other virological, pathological parameters. Then three specimens of cirrhotic patients whose health assessment results and laboratory data were similar and three normal liver specimens explanted from liver grafts prepared for liver transplantation were chosen for investigating gene expression of ECM and CAM using cDNA expression array. RESULTS: The cDNA array assay revealed 36.7% (36/96) of genes with changes, in which 26.3% (26/96) was up-regulated and 10.1% (10/96) was down regulated. Integrin (ITGA), collagen (COL), ADAMTS were identified as the characteristic changes of ECM and CAM gene expression levels. ITGA were demonstrated beta1 and beta2 sub-section changed in liver cirrhosis. CONCLUSION: ECM and CAM play an important role in the progression of liver cirrhosis after HCV infection. The capital mechanism is related to the inflammatory cells infiltration, the activation and transformation of ECM producing cells and the imbalance between production and elimination of ECM. PMID- 15810091 TI - Clinicopathological significance of heparanase and basic fibroblast growth factor expression in human esophageal cancer. AB - AIM: Human heparanase is an endo-D-glucuronidase that degrades heparan sulfate/heparin and has been implicated in a variety of biological processes. The objective was to investigate the expression of heparanase (Hps) and basic fibroblast growth factor (bFGF) and their relationship to neoangiogenesis and metastasis of human esophageal carcinoma. METHODS: Seventy-nine patients who had undergone esophageal resection for esophageal carcinoma without preoperative treatment were included in the present study. Immunohistochemistry was used to study the expression of Hps, bFGF and microvessel density (MVD) in 79 cases of esophageal carcinoma. bFGF and Hps were quantitatively detected with immunohistochemistry in 79 cases of human esophageal carcinoma and 19 cases of adjacent normal human esophageal carcinoma. Cd34 was used to explore the MVD as a marker of endothelial cells. RESULTS: Hps and bFGF expression in tumor tissue, being remarkably higher than that in normal esophageal tissue, were significantly correlated with clinicopathological features (depth of invasion, lymph-node metastasis and TNM stage) and MVD. CONCLUSION: The results of this study suggest that the coexpression of Hps and bFGF plays a key role in angiogenesis, invasion and metastasis of esophageal carcinoma. Hps and bFGF may serve as a predictor of progression in esophageal carcinoma. The expression of heparanase in esophageal carcinoma enhances growth, invasion, and angiogenesis of the tumor, and bFGF seems to be a potent antigenic factor for esophageal carcinoma. PMID- 15810092 TI - Relationship between retinopathy and cirrhosis. AB - AIM: To evaluate ophthalmic disorders with special attention to retinopathy in cirrhotic patients. Vitamin A deficiency-related ophthalmopathy, xerophthalmia, and color blindness may be documented in cirrhosis due to various etiologies. Retinopathy is an obscure feature of cirrhosis. METHODS: Thirty-two cirrhotic patients, who were followed up by Clinics of Gastroenterology, Izmir Ataturk Teaching and Research Hospital, were enrolled to the study. Associated systemic diseases such as diabetes mellitus and hypertension were excluded. Thirty-two healthy volunteers took part as the control subjects. All participants had ophthalmologic examination in the same hospital. RESULTS: Five (15.6%) of the cirrhotic subjects had soft exudate in the retina. None of the control subjects had retinopathy (P<0.05). Intraocular pressure (IOP) measured for both eyes were also significantly lower in the cirrhotics (P<0.05 vs P = 0.01). There were no statistically significant differences between the two groups in terms of other ophthalmic pathologies. The ophthalmic findings did not show up any differences according to the etiology of cirrhosis. CONCLUSION: Soft exudates may develop in cirrhotic patients probably due to loss of synthetic function of liver and hemodynamic effects of portal hypertension. Retinopathy must be sought in cirrhosis because of its severe morbidity. PMID- 15810093 TI - Unusual clinical course of metachronous melanomas of the upper digestive system. AB - Melanoma of the gastrointestinal tract is a rare, highly malignant neoplasm of poor prognosis. This is description of an unusual case of surgically treated patient with two metachronous malignant melanomas of the stomach and the esophagus. The former lesion was located in the cardia and effectively treated with R0 total gastrectomy. The latter was recognized after 67 mo and appeared as irregular, flat, pigmented areas located in the mid esophagus. Subtotal esophagectomy via right-sided thoracotomy, laparotomy and left-sided cervicotomy was performed, but neoplastic cells were found in distal margin (R1). Fourteen months after esophagectomy multiple lung metastases were detected. Patient died 2 mo later. PMID- 15810094 TI - Etiological role of brucellosis in autoimmune hepatitis. AB - To show that brucellosis may trigger autoimmune hepatitis (AIH), in addition to nonspecific liver involvement and toxic hepatitis, due to a class effect of tetracycline family used for treatment. We present a female patient admitted to our hospital due to partially improved fatigue and elevated liver enzymes following doxycycline and streptomycin usage for brucellosis. Brucellosis is endemic in our country, Turkey. It may involve any organ in the body. Liver is frequently involved. Doxycycline used for treatment occasionally may lead to hepatotoxicity. AIH is a necroinflammatory disease of the liver. Certain drugs (e.g. minocycline), toxins, and viruses (hepatitis B, hepatitis C, EBV, etc.) can trigger AIH. Only one case of AIH probably caused by doxycycline and brucellosis was reported. We discuss the relationship between brucellosis, AIH, and hepatotoxicity of doxycycline. Brucellosis may trigger AIH. PMID- 15810095 TI - Extraskeletal myxoid chondrosarcoma metastatic to the pancreas: a case report. AB - Extraskeletal myxoid chondrosarcoma (EMC) is a low-grade sarcoma characterized by developing metastases and local recurrence in high rate. It is mainly deep seated in the proximal extremities. The most common metastatic sites are the lungs, soft tissues, lymph nodes, bones and the brain. To our knowledge, no case of clearly defined EMC has been reported to date developing a metastasis in the pancreas. We describe a case of a man suffering from EMC who developed a single pancreatic metastasis 20 years after the initial diagnosis. A 49-year-old man was submitted to surgical excision of an EMC, in left thigh, 20 years ago. Fourteen years after the initial diagnosis a local recurrence in left thigh occurred. Multiple lesions of metastatic origin, in both lungs, were excised via thoracotomies until the time being. In 2003, as a part of a periodically performed imaging control, an abdominal CT scan was performed revealing a solid lesion in the pancreas. Distal pancreatectomy was performed. The histopathology of the excised specimen proved to be the one of metastatic lesion of EMC. The above-mentioned case of EMC is, as far as we know, the first one described developing a certain pancreatic metastasis. PMID- 15810096 TI - Extrahepatic portal vein aneurysm: two case reports of surgical intervention. AB - We report two cases of extrahepatic portal vein aneurysm, and both of them underwent surgical intervention. The first case had a mild pain in right upper quadrant of the abdomen; the second had no obvious symptoms. Physical examination revealed nothing abnormal. Both of them were diagnosed by magnetic resonance imaging angiography (MRA). One of the aneurysms was located at the main portal vein, the other, at the confluence of the superior mesenteric vein and the splenic vein, and these two places are exactly the most common locations of the extrahepatic portal vein aneurysm reported in the literature (30.7% each site). The first case underwent aneurysmorrhaphy and the second case, aneurysm resection with splenectomy. Both of them recovered soon after the operation, and the symptom of the first case was greatly alleviated. During the follow-up of half a year, no complication and adverse effect of surgical intervention was found and the color Doppler ultrasonography revealed no recurrence of the aneurysmal dilation. We suggest that surgical intervention can alleviate the symptom of the extrahepatic portal vein aneurysm and prevent its complications effectively and safely for low risk patients. PMID- 15810097 TI - Extensor carpi ulnaris subluxation or dislocation? Ultrasound measurement of tendon excursion and normal values. AB - Dislocation or subluxation of the extensor carpi ulnaris (ECU) tendon is one of the differential diagnoses of ulnar-sided wrist pain. No data exists on the degree of subluxation occurring in normal wrists. Retrospective review of surgical cases revealed two patients with dorsal and four with volar instability. Magnetic resonance imaging and ultrasound were not always predictive of the direction of instability. A standardised ultrasound technique was used to evaluate ECU subluxation in 20 asymptomatic wrists. In normal subjects, as a percentage of the width of the osseous groove, the volar edge of extensor carpi ulnaris may displace by up to 40% beyond the volar lip of the osseous groove with wrist flexion and the dorsal edge by 33% beyond the dorsal lip with wrist extension. The effect of gender on normal range and the diagnostic use of the standardised ultrasound technique need further evaluation. PMID- 15810098 TI - Kienbock disease treated by excisional arthroplasty with a palmaris longus tendon ball: a comparative study of cases with or without bone core. AB - We hypothesised that using a palmaris longus tendon ball (PLTB) with bone core (w bc) after excisional arthroplasty for Kienbock disease would maintain post operative carpal height compared to a PLTB without bone core (w/o bc). Seventeen hands of 16 consecutive patients with Kienbock disease at Lichtman stage IIIA or IIIB were treated by replacement of the lunate with a PLTB w bc or w/o bc. We evaluated the clinical and radiological outcomes at one, three and 12 months after surgery. According to Dornan and Lichtman criteria respectively, there were no significant differences between the two groups. In the w bc group, the post operative values of the carpal height ratio (CHR) were maintained at the same level as pre-operative values for one year, while the post-operative CHR values in the w/o bc group were significantly lower than those in the w bc group. Our results indicate that in Kienbock disease, arthroplasty using a PLTB w bc can maintain CHR at one year after surgery compared to arthroplasty using a PLTB w/o bc. PMID- 15810099 TI - The influence of motor axon misdirection on muscle contraction force in early nerve repair in a rat sciatic nerve model. AB - We examined the influence of misdirection of regenerating motor axons toward the distal sensory Schwann tubes on the muscle contraction force in early nerve repair using a rat sciatic nerve model. At 0, 1, 2, 4 and 8 weeks after severing the tibial, peroneal and sural nerves, the proximal stump of the tibial nerve was anastomosed with the distal stumps of both the peroneal and sural nerves using tubulisation (n=10 in each of five groups). We intentionally used the distal stump of the sural nerve (a sensory nerve) to induce regeneration in motor axons from the proximal tibial nerve stump toward the distal sensory nerve stump. Twenty-four weeks after nerve repair, isometric contraction force and wet weight of the anterior tibial muscle were measured, and the numbers of regenerated myelinated axons (motor and sensory) in the distal sural and peroneal nerves were counted. The rates of sural nerve regeneration were significantly higher at weeks 0 and 1 than at the later repair times. However, muscle contraction force and muscle wet weight did not differ significantly between groups in which nerves were repaired within four weeks of severance. These results indicate that peripheral nerve repair within four weeks of severance does not influence the muscle contraction force of single muscle despite the misdirection of motor axons toward the distal sensory Schwann tubes. PMID- 15810101 TI - Results of endoscopic management of primary versus recurrent carpal tunnel syndrome in long-term haemodialysis patients. AB - In long-term haemodialysis patients, carpal tunnel syndrome (CTS) frequently occurs as a result of amyloid deposition, originating from beta-2 microglobulin, to the flexor retinaculum, paratenons and tendons themselves, which leads to an increase in carpal canal pressure and compression of the median nerve. Surgical procedures can rectify the condition, but continuing maintenance haemodialysis sometimes causes recurrence. We endoscopically operated 1848 hands primarily, 104 recurrent post-endoscopic procedure hands and 130 recurrent post-open procedure hands using the Universal Subcutaneous Endoscope (USE) system, then analysed clinical symptoms and electrophysiological recovery for more than six months post operatively. The patients were satisfied with the clinical results. Optimal electrophysiological improvements were reported. There were no statistical differences between three groups, except in recovery of touch sensation, which was better in the post-endoscopic group than in the post-open group. There were no complications in this series. Our minimally invasive endoscopic procedure, using the USE system, is safe and effective for primary and recurrent CTS in haemodialysis patients. PMID- 15810100 TI - Arthroscopic resection of wrist ganglia. AB - The arthroscopic resection of synovial cysts of the wrist is a simple technique which is comfortable for the patient. We report on a series of 96 patients with dorsal synovial cysts (75 women, 21 men). All patients had undergone preliminary treatment which had been unsuccessful. We operated on 32 patients with a volar cyst (27 women, five men). All the patients were operated on as outpatients under local regional anaesthesia. For the dorsal cysts, after having precisely located the cyst, it is then resected after having inserted a shaver directly through the wall of the cyst starting with the capsule. For the volar cysts the arthroscope was inserted through a 3-4 portal and the shaver was inserted through a 1-2 radiocarpal portal. In all cases, there was no immobilisation and a range of motion was started the same day. For the dorsal cysts, our average follow-up was 34 months (range 12-46 months). There were no complications. We had four recurrences. For the palmar cysts, our average follow-up was 26 months (range 12 39 months). There have been no recurrences to date. PMID- 15810102 TI - Outcome of surgery in patients with painful dorsal wrist ganglia and arthroscopic confirmed ligament injury: a five-year follow-up. AB - We report the outcome of a five-year follow-up after wrist arthroscopy and excision of painful dorsal wrist ganglia. The findings at the time of surgery have previously been published. Patients responded to a validated postal questionnaire regarding ganglion recurrence, wrist pain and function. None of the responding patients had recurrence of the ganglia since surgery but only one patient had remained pain free with normal function following surgery. Three of the remaining patients reported moderate to severe problems with work and four reported minimal work problems. Our findings suggest patients with arthroscopic confirmed ligament injuries leading to joint instability or localised osteoarthritis may develop functional disability but less severe injuries are unlikely to cause persistent problems in the short- to medium-term. Surgical excision of the ganglion can give lasting satisfactory cosmetic outcome despite persisting underlying ligament pathology. PMID- 15810103 TI - Radiographic morphology of the ulnar head. AB - Posteroanterior radiographs of 102 normal wrists were studied to determine the morphological characteristics of the ulnar head associated with the value of ulnar variance. Ulnar variance, ulnar styloid length, ulnar head diameter, ulnar seat inclination, and the distance of ulnar head peak to the distal radioulnar joint were measured together with the third metacarpal length as a reference of the size of the wrist. Moderate correlation was confirmed not only between ulnar variance and ulnar seat inclination but also between ulnar variance and the distance of the ulnar head peak. Furthermore, we found a statistically significant correlation between ulnar variance and ulnar head diameter. The results showed that whole ulnar head configurations are affected by ulnar variance although there are considerable variations. PMID- 15810104 TI - Dorsal plating for comminuted intra-articular fractures of the distal end of the radius. AB - Forty-one dorsally displaced intra-articular fractures of the distal radius were treated by open reduction and internal fixation with the dorsal Symmetry plates. The average age at the time of the injury was 49 years. An average follow-up period was 15 months. The final radial length averaged 11 mm, radial inclination 23 degrees, volar tilt 7 degrees, ulnar variance 1.5 mm, and articular incongruity 0.3 mm. According to the Gartland and Werly scales, 36 fractures were excellent and five were good. However, ulnar variance increased more than 3 mm during follow-up in eight patients, and volar tilt increased more than 5 degrees during follow-up in ten patients. Use of dorsal Symmetry plate is effective for unstable comminuted intra-articular distal radius fractures, but severely comminuted fractures may possibly undergo re-displacement post-operatively. PMID- 15810105 TI - Superficial branch of the radial nerve piercing the brachioradialis tendon to become subcutaneous: an anatomical variation with clinical relevance. AB - We dissected 20 preserved Caucasian cadaveric upper limbs looking at the relation of the superficial branch of the radial nerve (SBRN) to the brachioradialis tendon. SBRN emerged from deep to superficial position by piercing the brachioradialis tendon near its dorsal border in four limbs. The resulting dorsal tendinous band compressed the nerve and prevented longitudinal gliding movement during ulnar flexion. This is likely to increase the risk of chronic compression neuropathy (Wartenberg's syndrome). In two of these four limbs, there was a communication between the SBRN and lateral cutaneous nerve of the forearm. No such communication was found in the remaining 16 forearms. This communication could contribute to the minimal area of sensory loss observed in Wartenberg's syndrome. We recommend that this anatomical anomaly is looked for and if present dealt with during surgical treatment of Wartenberg's syndrome, as it is likely to predispose to chronic compression neuropathy. PMID- 15810106 TI - Differences in radiographic findings between scaphoid fracture patterns. AB - Scaphoid fracture patterns can be divided into volar and dorsal type using three dimensional computed tomography. Ninety-nine patients underwent this examination plus radiography to determine how often fracture pattern was identifiable by radiography including proximal fragment ratio (PFR) measurement. Oblique radiographs obtained at 45 degrees of pronation demonstrated a volar displacement in 28 of 37 volar type fractures in three-dimensional computed tomography (76%), while as oblique view with 45 degrees of supination could identify the dorsal type in ten of 18 (56%). PFR showed a significant difference, with means of 0.59 for volar type and 0.43 for dorsal type. Using oblique views in the two rotational directions together with the PFR, the two fracture patterns could be differentiated in 35 of 43 fractures (81%). No differences were evident between the two fracture patterns with respect to prevalence of dorsal intercalated segment instability deformity. PMID- 15810107 TI - Radiocarpal arthrodesis for osteoarthritis following fractures of the distal radius. AB - To compare the radioscapholunate (RSL) arthrodesis and radiolunate (RL) arthrodesis as a treatment for radiocarpal osteoarthritis following fractures of the distal radius, nine patients, 23 to 70 years old (average 41) at the time of surgery, were assessed two to 33 years after surgery. The periods between injury and surgery ranged from four months to 30 years. RSL arthrodesis was performed in three cases and RL arthrodesis in six. Post-operative wrist pain disappeared in six and was decreased in the other three. In the RSL group, the total arc of wrist flexion and extension was reduced from 50 degrees pre-operatively to 35 degrees post-operatively. In the RL group, it was increased from 72 degrees to 76 degrees after surgery. Grip strength improved in most patients, from 7 to 18 kg in the RSL group, and from 16 to 27 kg in the RL group. On roentogenogram, three patients showed arthritic changes in the adjacent joints, but there were no symptoms in two of the three patients. We concluded that partial radiocarpal arthrodesis (preferably RL arthrodesis) is a reliable procedure for radiocarpal osteoarthritis following fractures of the distal radius. PMID- 15810109 TI - Traumatic extrusion and replacement of autoclaved diaphyseal segment of radius: a case report. AB - A 17-year-old patient had a traumatic extrusion of 10 cm distal radial segment. The segment of bone was replaced after cleaning and autoclaving. At four years follow-up, there was complete incorporation with almost full functional recovery. We report this case for its rarity and successful result. PMID- 15810108 TI - Flap reconstruction following high pressure injection injuries of the hand: a report of three cases. AB - High pressure injection injuries are well known to cause significant injury to the hand, with high amputation rates and poor functional outcome. Surgical treatment consists of early aggressive debridement followed by secondary closure. Flap reconstruction is a reconstruction option which can increase the chance of digit salvage, as well as give an acceptable functional and cosmetic result. We review three cases of flap reconstruction following high pressure injection injuries, and discuss their role in the treatment of these injuries. PMID- 15810110 TI - Suicidal mercury injection into the upper limb: a case study. AB - We report a rare case of self-injection of mercury into the subcutaneous tissue of the upper limb. A multi-disciplinary management approach was adopted including cooperation between toxicologists, orthopaedic surgeons, radiologists and environment safety personnel. Surgical removal of mercury under radiological screening and systemic intoxication treated by chelating agents, namely dimercaprol and succimer. Serial serum and urine mercury levels showed an initial rise despite surgical removal and returned to normal after a prolonged period of time. Safety precautions were taken during surgery to avoid inadvertent intoxication of staff. Contamination of the operation theatre was monitored by the amount of mercury vapour released into the air. All personnels involved in the management of the patient did not show any evidence of mercury intoxication. PMID- 15810111 TI - Vascular malformation of hand: cavernous lymphangioma. AB - A case of congenital cavernous lymphangioma of the hand presenting in a newborn is presented. The mass on the dorsum of the hand was removed with the overlying skin and underlying paratenon at three years of age. Re-surfacing was performed with a free flap transfer. There were no post-operative complications during one year of follow-up. PMID- 15810112 TI - "Life" above the neck: brain tumour presenting as carpal tunnel syndrome. AB - Carpal tunnel syndrome is a common condition with many aetiologies. We present a case report of a glioblastoma which presented as carpal tunnel syndrome, and highlights that the most proximal site for pathology is not the spinal cord. PMID- 15810113 TI - Endoscopic carpal tunnel release for recurrent carpal tunnel syndrome after previous open release. AB - Recurrent carpal tunnel syndrome from various causes has been shown to occur in up to 19% of patients. Endoscopic carpal tunnel release has been used to decompress the median nerve in carpal tunnel syndrome for many years. However, endoscopic release for recurrent carpal tunnel syndrome after previous surgical release has not been reported. Nine hands in six patients had recurrent carpal tunnel syndrome five to 20 years after previous open carpal tunnel release. All the cases were successfully treated with endoscopic release. PMID- 15810114 TI - Synovial chondromatosis of the distal radioulnar joint. AB - Synovial chondromatosis is a rare lesion in the wrist and it is extremely uncommon in the distal radioulnar joint. This case presented with wrist pain, swelling and locking secondary to synovial chondromatosis of distal radioulnar joint. PMID- 15810115 TI - Dorsal pentagonal island flap: a technique of web reconstruction for syndactyly that facilitates direct closure. AB - Though many techniques have been described for the correction of syndactyly, current methods result in skin deficiency that requires skin grafting, especially at the finger bases. We discuss the technical aspects and document our experience with the dorsal pentagonal metacarpal island flap for reconstruction of the web commissure in a series of 12 patients (22 webs) with syndactyly of varying complexity. In appropriately selected cases, this technique can improve reconstruction of the web commissure, facilitating direct closure, minimising the need for skin grafts and offers the potential for continued growth. This reduces operative time significantly and simplifies post-operative wound care. In this series, all our cases healed primarily in two to three weeks with minimal donor site morbidity. There were no post-operative complications, although the prominent dorsal scar remains a concern. At an average follow-up of 33.7 months, no cases of contracture or web creep after correction were noted. PMID- 15810116 TI - Irreducible palmar dislocation of the proximal interphalangeal joint of a finger evaluated by magnetic resonance imaging: a case report. AB - The irreducible palmar dislocation of the proximal interphalangeal joint (PIPJ) of a finger is rare and central slip rupture with the interposition of the lateral band in the PIPJ is extremely rare. In our present case, magnetic resonance imaging (MRI) enabled a very effective and detailed evaluation of soft tissue damage. PMID- 15810118 TI - High blood pressure cases soar. PMID- 15810117 TI - Late psychosocial effects of congenital hand anomaly. AB - Lifetime-standing psychosocial effects of congenital hand anomalies are inevitable in patients who have not received a comprehensive treatment with appropriate timing and approach. Herein, two adult cases of untreated thumb polydactyly are presented. Both of them had hands with striking appearance and late consequent psychosocial problems. PMID- 15810119 TI - Arsenic, mercury and lead taint some herbal products. PMID- 15810120 TI - Ask the doctors. I have been diagnosed with a poor ejection fraction from my left ventricle. What does that mean, and what medications would be most beneficial? PMID- 15810121 TI - Ask the doctors. Five years after I had a bypass operation, I developed chest pains, and my doctor implanted a stent that wasn't coated with any drug. After two months, the chest pain returned, and I was given radiation treatment. Now after a year, I still feel fine. Is radiation a long-term preventive treatment? PMID- 15810122 TI - Contribution of metabolic factors to alanine aminotransferase activity in persons with other causes of liver disease. AB - BACKGROUND & AIMS: Nonalcoholic fatty liver disease has been defined by the presence of hepatic steatosis in the absence of other chronic liver diseases. We sought to determine whether obesity, insulin resistance, and the metabolic syndrome, which are the main risk factors for nonalcoholic fatty liver disease, are associated with similar elevations in serum alanine aminotransferase activity in persons with and those without other causes of chronic liver disease. METHODS: Adult participants of the third National Health and Nutrition Examination Survey were divided into those with causes of chronic liver disease (n = 1037), defined as viral hepatitis, excessive alcohol consumption, or increased transferrin-iron saturation, and those without (n = 8004). RESULTS: Among persons with other causes of chronic liver disease, obesity (adjusted odds ratio, 4.9; 95% confidence interval, 2.5-9.4), insulin resistance (adjusted odds ratio, 6.8; 95% confidence interval, 3.0-15.5, comparing the highest and the lowest quartile), and the metabolic syndrome (adjusted odds ratio, 3.3; 95% confidence interval, 1.4-8.0) were all strongly associated with increased alanine aminotransferase activity (>43 IU/L). Among persons without other causes of chronic liver disease, statistically similar associations were identified. CONCLUSIONS: Obesity, insulin resistance, and the metabolic syndrome are strong predictors of increased alanine aminotransferase activity in the US population, both in persons with and in persons without other causes of chronic livers disease. We hypothesize that metabolic fatty liver disease related to these conditions is the cause of the increased alanine aminotransferase activity and may be underrecognized in persons with other causes of chronic liver disease. PMID- 15810123 TI - Bibliography. Current world literature. Cancer biology. PMID- 15810124 TI - The learning curve measured by operating times for laparoscopic and open gastric bypass: roles of surgeon's experience, institutional experience, body mass index and fellowship training. AB - BACKGROUND: Surgeons must overcome a substantial learning curve before mastering laparoscopic Roux-en-Y gastric bypass (LRYGBP). This learning curve can be defined in terms of mortality, morbidity or length of surgery. The aim of this study was to compare the learning curves in terms of surgical time for the first 3 surgeons performing LRYGBP in our hospital with the length of surgery for open gastric bypass (CONTROLS). METHODS: We compared 494 primary LRYGBPs performed by 3 surgeons (393 by 1st SURGEON, 57 by 2nd SURGEON and 44 by 3rd SURGEON) to 159 open vertical banded gastroplasty-Roux-en-Y gastric bypasses (CONTROLS). Data for LRYGBP patients were prospectively obtained. Factors that significantly affected the length of surgery were identified by univariate and multivariate linear regression analysis. RESULTS: LRYGBP and CONTROL patients were similar in age, height, weight and BMI, although more CONTROLS were male. Median time for the 1st SURGEON performing LRYGBP dropped for each subsequent 100 operations: 1st 100 - 190 min, 2nd 100 - 135 min, 3rd 100 - 110 min and 4th 100 - 100 min. Median time for 2nd SURGEON performing LRYGBP was 120 min, 3rd SURGEON 173 min and CONTROLS 64 min. Length of surgery significantly correlated with surgical experience in terms of numbers of operations and BMI of patient. Times for 2nd SURGEON, a fellowship trained laparoscopic surgeon, started significantly faster than 1st SURGEON's, but did not significantly improve with experience. 3rd SURGEON's initial times were similar to 1st SURGEON's, but his times improved more rapidly with experience. Times for CONTROLS were significantly faster than all laparoscopic groups and did not correlate with operation number or patient BMI. CONCLUSIONS: The length of surgery for LRYGBPs continued to shorten beyond 400 operations for the first surgeon performing LRYGBP in our hospital. Previous fellowship training in LRYGBP shortened surgical times during initial clinical experience as an attending for the second surgeon. The learning curve was truncated because of the already established LRYGBP program. PMID- 15810125 TI - Perspectives. MedPAC seeks level payment playing field for competition, P4P. PMID- 15810126 TI - Southern Africa: reports document legal responses to HIV/AIDS. AB - A series of reports have been issued on the legal response to HIV/AIDS in eight member states of the Southern African Development Community (SADC). The states are Botswana, Malawi, Mozambique, Namibia, South Africa, Swaziland, Zambia, and Zimbabwe. PMID- 15810127 TI - NGO Code of Practice released. AB - Renewing Our Voice: Code of Good Practice for NGOs Responding to HIV/AIDS was launched on World AIDS Day 2004, with the endorsement of over 150 NGOs from around the world. Eleven NGOs collaborated on the project. PMID- 15810129 TI - Nigeria addresses HIV/AIDS in the workplace. AB - In 2004 Nigeria undertook several important steps to address issues related to HIV/AIDS and the workplace. PMID- 15810128 TI - Thai Drug Users Network receives human rights award. AB - The 2004 International Award for Action on HIV/AIDS and Human Rights has gone to the Thai Drug Users' Network (TDN). PMID- 15810130 TI - New report on the impact of HIV/AIDS on the world of work. AB - A new report presents global estimates of the impact of HIV/AIDS on the labour force, on men and women of working age, on children, and on the economy in 50 countries, and traces its possible future impact in projections to 2015. PMID- 15810131 TI - Prison needle exchange programs work. AB - On 27 October 2004 the Canadian HIV/AIDS Legal Network and the Ontario Medical Association (OMA) released reports recommending that needle exchange programs (NEPs) be implemented in prisons, based on evidence that such programs promote and protect the health of both prisoners and prison staff. At the same time, the Scottish Prison Service announced that it was looking at providing such programs, and an Iranian physician reported on a pilot project in his country. PMID- 15810132 TI - Is the world finally waking up to HIV/AIDS in prisons? A report from the XV International AIDS Conference. AB - Issues related to HIV/AIDS in prisons have traditionally received little attention at the International AIDS Conference, despite the fact that levels of HIV prevalence within prison populations tend to be much higher than in the general population. However, the conference in Bangkok may represent a turning point. This article highlights some of the relevant findings presented at the conference. A longer version of this article, including background information about HIV/AIDS in prisons worldwide, has been published in the September 2004 issue of Infectious Diseases in Corrections Report. PMID- 15810133 TI - Canada: study provides further evidence of risk of hepatitis C and HIV transmission in prisons. AB - In 2002, as part of a larger injection drug use social network study undertaken in Winnipeg, a study was undertaken at Stony Mountain Institution to obtain data on the social network interactions in the prison and the potential role these interactions have on facilitating or inhibiting the spread of bloodborne pathogens among injection drug users. The study, although small in size and largely exploratory, provides further evidence of the risk of HCV and HIV transmission in prisons and its potential public health implications. PMID- 15810134 TI - Canadian court orders remand centre to provide prescribed medication and permit consultation with outside physician. AB - In two applications brought by Geary, a provincial prisoner, the Alberta Court of Queen's Bench ordered the Edmonton Remand Centre to provide Geary with prescribed medication and to permit him to consult an outside physician. In each application, Geary was asking the court on an urgent basis to order interim measures until such time as the court could hear his full application for release from custody. This decision has important implications for prisoners living with HIV/AIDS, some of whom report receiving medical treatment that does not meet professionally accepted standards. It is also significant that, in finding for the prisoner, the court refers to the Charter, the United Nations Standard Minimum Rules for the Treatment of Prisoners, and American constitutional case law. PMID- 15810135 TI - Canadian court denies prisoner access to anti-anxiety medication. AB - In a judgment released on 22 September 2004, the Federal Court refused to order a federal correctional institution to provide a prisoner with the anti-anxiety medication clonazepam, one among a class of medications known as benzodiazepines. Benzodiazepines are available for prescription by Correctional Service Canada (CSC) physicians and psychiatrists. However, the CSC formulary provides for a discontinuation schedule for inmates who come from the provincial penitentiary system where the continued use of benzodiazepines is not indicated. PMID- 15810136 TI - Prisoner denied access to medical marijuana. AB - In a case that has recently come to the attention of the editors, the Federal Court refused to order Health Canada to provide a federal prisoner with medical marijuana, even though he possessed legal authorization to possess marijuana. PMID- 15810137 TI - US: study links incarceration and HIV rates in black communities. PMID- 15810138 TI - Canada: prison networking and research & advocacy groups founded. PMID- 15810139 TI - Court orders Immigration and Refugee Board to take into account evidence of HIV related discrimination. AB - On 31 August 2004, the Federal Court granted an HIV-positive woman's application for judicial review of a decision by the Immigration and Refugee Board (IRB). The issue before the court was whether the Board erred by ignoring documentary evidence before it with respect to the treatment of people living with HIV/AIDS in Nigeria. The court ordered that a newly constituted panel redetermine the claim in light of evidence of the discrimination faced by HIV-positive people in Nigeria. PMID- 15810140 TI - [Questionnaire study of general practitioners regarding experience and attitude to opioid substitution therapy]. AB - General practitioners (GPs) in their surgeries and substitution treatment centres are the major providers of opioid maintenance treatment in a number of European countries. Although in the Czech Republic any GP has been allowed to prescribe buprenorphine (Subutex) since 2001, the opioid substitution treatment provided by primary care professionals has not been the subject of research to date. OBJECTIVE: To collect and analyze data on GPs' experience gained with opioid maintenance treatment in the Czech Reupblic, their attitudes and needs. METHOD: A structured questionnaire was distributed via the Bulletin of the Association of General Practitioners and district Association representatives. The validity of study results may be affected by a low respondence rate (10%) with 398 questionnaires only returned by mail. RESULTS: Twenty-eight (7%) GPs reported to have gained experience with buprenorphine which was most frequently prescribed in the regions with the highest prevalence of heroin users, i.e. in Prague and the Usti nad Labem region (27% and 12%, respectively). Other regions, including wes- tern and southern Bohemia with relatively high prevalence of heroin users, showed lower buprenorphine prescription rates (0-6%). Most buprenorphine prescribers (78%) rated their experience as positive or highly positive. Availability and effectiveness were seen as the main pros of the substitution treatment. One third of the GPs who have not prescribed opioid maintenance treatment yet are considering doing so in the future. Greater awareness of drug abuse issues and availability of methodical guidance and consulting in opioid substitution treatment are going to become the most relevant factors in the future. Possible reportability of data on opioid maintenance treatment to a central registry does not seem to be a major obstacle to implementing the substitution treatment in the GPs' surgeries. Decision makers should take advantage of the GPs' potential to promote the opioid maintenance treatment in the Czech Republic. PMID- 15810141 TI - Abstracts of the Vth International Conference on Cancer-Induced Bone Disease. Davos, Switzerland, 20-24 March 2005. PMID- 15810142 TI - [Abstracts of the VII Magyar Neuropsychopharmacology Congress. Tihany, Hungary, September 30-October 2, 2004]. PMID- 15810143 TI - Abstracts of the European College of Neuropsychopharmacology Workshop on Neuropsychopharmacology for Young Scientists in Europe. Nice, France, March 3-6, 2005. PMID- 15810145 TI - [The 78th annual meeting of the Japanese Society for Bacteriology. Tokyo, Japan. April 4-6, 2005. Abstracts]. PMID- 15810146 TI - Abstracts of the 2nd SERI-ARVO Meeting on Research in Vision and Ophthalmology. 16-20 February 2005, Singapore. PMID- 15810144 TI - Transcript AA454543 is a novel prognostic marker for hepatocellular carcinoma after curative partial hepatectomy. AB - BACKGROUND: We have previously reported on the cDNA microarray gene expression profiles of hepatocellular carcinomas (HCCs). Among the genes that show prognostic significance and are overexpressed in tumor compared with adjacent nontumorous liver, transcript AA454543 may have potential for practical use. Our aim is to validate the prognostic significance of transcript AA454543 by alternative research methods and in a separate group of HCC patients. METHODS AND RESULTS: The data of transcript AA454543 derived from microarray analysis of 48 patients having curative partial hepatectomy (group 1) were verified by quantitative reverse transcription polymerase chain reaction (r = 0.618, P < .001). A separate sample set of HCCs obtained from 53 patients (group 2) was examined and the association of AA454543 expression level with overall survival was again validated (P = .027). By Cox regression analysis, transcript AA454543 [hazard ratio (HR) = 3.0, P = .017] and pathologic tumor node metastasis (pTNM) stage (HR = 3.3, P = .010) were independent prognostic factors for overall survival. The accuracy of prediction for 3-year overall survival for transcript AA454543 (74.2%, P = .001) and pTNM stage (76.4%, P = .001) was comparable as measured by the area under the receiver operating characteristic curve. CONCLUSION: Transcript AA454543 is potentially useful molecular prognostic marker for overall survival after curative partial hepatectomy for HCC. PMID- 15810147 TI - Abstracts of the American Society of Andrology 30th Annual Meeting. March 30 April 5, 2005, Seattle, Washington, USA. PMID- 15810148 TI - [Abstracts of the 46th Congress of the German Society of Pneumology. 16-19 March 2005. Berlin, Germany]. PMID- 15810149 TI - [Pathology 2004. 15-19 November 2004, Paris, France. Proceedings and abstracts]. PMID- 15810150 TI - [The 45th annual meeting of the Japanese Respiratory Society. Chiba City, Japan. April 14-16, 2005. Program and Abstracts]. PMID- 15810151 TI - Abstracts of the 78th Annual Meeting of the Japanese Pharmacological Society. March 22-24, 2005, Yokohama, Japan. PMID- 15810152 TI - [First Congress of the Spanish Federation of Societies of Nutrition, Food and Diet. Madrid, Spain, 9-11 March 2005]. PMID- 15810153 TI - Abstracts of the 2005 AAHPERD (American Alliance for Health, Physical Education, Recreation and Dance) National Convention. Chicago, Illinois, USA, April 12-16, 2005. PMID- 15810154 TI - Retraction of "The prognostic and predictive value of immunohistochemically detected HER-2/neu overexpression in 361 patients with ovarian cancer: a multicenter study" [Gynecol Oncol 2004;95:89-94]. PMID- 15810155 TI - Loss of cell-surface heparan sulfate expression in both cervical intraepithelial neoplasm and invasive cervical cancer. AB - OBJECTIVE: Syndecan-1 binds to various extracellular matrix components via its heparan sulfate glycosaminoglycans (HS-GAG) and most of its biological functions are considered to be associated with this process. The aims of this study are to investigate its expression in cervical neoplasms. METHODS: We investigated the expression of both the syndecan-1 core protein and cell-surface HS-GAG by immunohistochemistry in 53 cervical intraepithelial neoplasm (CIN), 19 microinvasive, 143 invasive cervical cancers, and 29 metastatic lymph node samples, and analyzed correlations with various clinicopathological features. RESULTS: The progression of CIN to early invasive cancer was found to associate with reduced levels of both syndecan-1 and HS-GAG expression. In squamous cell carcinomas, HS-GAG expression was significantly lower in cases with lymph vascular space invasion. Additionally, the overall survival rates for patients exhibiting low HS-GAG expression was significantly lower than patients exhibiting high HS-GAG expression (P = 0.019). Low HS-GAG expression in positive nodes was determined to be a disease-free and overall survival prognostic factor (P = 0.028 and P = 0.018, respectively). CONCLUSION: The loss of syndecan-1 and HS-GAG expression is an early event in cervical carcinogenesis. The loss of HS-GAG expression particularly in positive nodes can serve as an indicator of aggressive disease potential and poor prognosis in patients with invasive cervical cancer. PMID- 15810156 TI - Biological effects of indigenous medicinal plants Curcuma longa and Alpinia galanga. AB - The ethanolic extracts of Curcuma longa and Alpinia galanga exhibited excellent (100%) phytotoxic activity against Lemma minor. These extracts were also found to possess good antifungal activities against Trichophyton longifusus (65% and 60%, respectively), while in the brine shrimp lethality bioassay were found to be toxic with LD50 of 33 and 109 mincrog/ml, respectively. These extracts were found quite inert in antibacterial bioassay, while the extract from C. longa, tested for insecticidal activity, was also found to be devoid of any activity. PMID- 15810158 TI - Proceedings of the International Conference BIOCOMP 2002: Topics in Biomathematics and Related Computational Problems at the Beginning of the Third Millennium. PMID- 15810157 TI - [Radiological case study. Lymphoblasts]. PMID- 15810159 TI - Haemorheological alterations in Behcet's disease are not related to a tendency for venous thrombosis. AB - INTRODUCTION: Behcet's disease is associated with an increased risk of thrombosis, although the prothrombotic mechanisms are unclear. Alterations in blood rheology, particularly increased erythrocyte aggregation, might play a role in the development of such thrombotic events. MATERIALS AND METHODS: We measured plasma lipids, fibrinogen, haematocrit, erythrocite aggregation, erythrocyte deformability, blood viscosity, plasma viscosity and erythrocyte indexes in patients with a nonactive disease at sampling, and in a well-matched control group. The patient group comprised 42 patients with BD (21 male, 21 female aged 43+/-12 years) and the control group comprised 46 healthy volunteers (23 male, 23 female aged 45+/-13 years). Twelve of the 42 patients with BD had a previous documented history of deep vein thrombosis at least 6 months before entering the study, and the other 30 did not. RESULTS: Compared with controls, patients showed statistically higher fibrinogen concentrations (P=0.001), plasma viscosity (P=0.002), blood viscosity (P=0.006) and erythrocyte aggregation, both at stasis (P=0.001) and at a low shear rate (P=0.002): the other rheological parameters were not statistically significant. No differences were observed in the rheological parameters when patients with and without previous thrombotic episodes were compared. CONCLUSIONS: Although patients with BD show a moderate hyperviscosity syndrome, possibly related to chronic inflammation, this does not seem to play a role in the development of thrombotic events. PMID- 15810160 TI - Professor Halina Krzanowska. (1926-2004). PMID- 15810161 TI - Acute generalized exanthematous pustulosis (AGEP) induced by terbinafine with involuntary positive reintroduction. PMID- 15810162 TI - Ultrasound for central venous cannulation: economic evaluation of cost effectiveness. PMID- 15810163 TI - Ultrasound for central venous cannulation: economic evaluation of cost effectiveness. PMID- 15810164 TI - [Proceedings from the X Conference of Warsaw-Lodz Branch of the Polish Society of Nephrology]. PMID- 15810165 TI - [Proceedings of the III National Conference on the Prevention of Cardiovascular Disease. Rome, Italy, 15-16 April 2004]. PMID- 15810167 TI - [Proceedings of the National Congress of the Survival of the Kidney and the Transplanted Patient. Udine, Italy, 4 October 2002]. PMID- 15810166 TI - [Efforts in the individualization of the adjuvant systemic therapy in breast cancer--ASCO, 2004]. PMID- 15810168 TI - [Report from the 10th Congress of the German Radio-oncologists]. PMID- 15810169 TI - [Methodologic recommendations of the Oncology and Radiotherapy College. Protocol for oncology care. Diagnostic algorithms in the course of patient follow-up]. PMID- 15810170 TI - Computerized physician order entry systems: the right prescription? AB - Policymakers increasingly urge the use of information technology to improve the quality and efficiency of health care. In particular, computerized physician order entry (CPOE) is emphasized for its ability to reduce prescribing errors inherent in paper-based systems. This Issue Brief summarizes research that sounds a cautionary note about the potential for computerized systems to facilitate medication errors, as well as reduce them. PMID- 15810171 TI - Structure of hospital networks in California affects pricing. PMID- 15810172 TI - Inflammation-induced hyperexcitability of nociceptive gastrointestinal DRG neurones: the role of voltage-gated ion channels. AB - Gastrointestinal (GI) inflammation modulates the intrinsic properties of nociceptive dorsal root ganglia neurones, which innervate the GI tract and these changes are important in the genesis of abdominal pain and visceral hyperalgesia neurones exhibit hyperexcitability characterized by a decreased threshold for activation and increased firing rate, and changes in voltages-gated Na(+) and K(+) channels play a major role in this plasticity. This review highlights emerging evidence that specific subsets of channels and signalling pathways are involved and their potential to provide novel selective therapeutics targets for the treatment of abdominal pain. PMID- 15810174 TI - Effect of acute acoustic stress on anorectal function sensation in healthy human. AB - Little is known about the effects of acute acoustic stress on anorectal function. To determine the effects of acute acoustic stress on anorectal function and sensation in healthy volunteers. Ten healthy volunteers (7 M, 3 F, mean age 34 +/ 3 years) underwent anorectal manometry, testing of rectal compliance and sensation using a barostat with and without acute noise stress on separate days. Rectal perception was assessed using an ascending method of limits protocol and a 5-point Likert scale. Arousal and anxiety status were evaluated using a visual analogue scale. Acoustic stress significantly increased anxiety score (P < 0.05). Rectal compliance was significantly decreased with acoustic stress compared with control P (P < 0.000001). In addition, less intraballoon volume was needed to induce the sensation of severe urgency with acoustic stress (P < 0.05). Acoustic stress had no effect on hemodynamic parameters, anal sphincter pressure, threshold for first sensation, sensation of stool, or pain. Acute acoustic stimulation increased anxiety scores, decreased rectal compliance, and enhanced perception of severe urgency to balloon distention but did not affect anal sphincter pressure in healthy volunteers. These results may offer insight into the pathogenesis of stress-in-induced diarrhoea and faecal urgency. PMID- 15810175 TI - Involvement of NO in gastric emptying of semi-solid meal in conscious pigs. AB - The influence of non-selective nitric oxide synthase (NOS) inhibition on gastric emptying of a semi-solid meal was studied in conscious pigs. Antroduodenal motility and fundic compliance were also assessed to evaluate the mechanisms at the origin of potential alteration in gastric emptying pattern. N(G)-nitro-L arginine methyl ester (L-NAME; 20 mg kg(-1) i.v.) delayed gastric emptying (half emptying time of 128.98 +/- 16.86 min vs 73.74 +/- 7.73 min after saline, P < 0.05, n = 6) as a result of decreased proximal gastric emptying. No changes were observed for distal gastric emptying as a result of unchanged antral motility. Similarly, no changes were noted on duodenal motor patterns either in the fasted or in the fed state. L-NAME decreased fundic compliance in fasted state (49 +/- 11 mL mmHg(-1) vs 118 +/- 15 mL mmHg(-1) after saline, P < 0.05, n = 6). As this phenomenon is expected to increase emptying rate, the gastroparesis induced by NOS inhibition is thus likely to originate from distal resistive forces. It is concluded that NO positively modulates gastric emptying. PMID- 15810176 TI - Did I make the grade? Ethical issues in psychosocial screening of children for adoptive placement. AB - This article suggests that psychological evaluations as a "screen for normalcy" with a view to adoption are a breach of the child's right to psychological integrity and privacy under international human rights law. A foreseeable outcome of such psychological screens, especially for the older foster child who has experienced multiple placements, is an unreliable mental health diagnosis. Normal, albeit maladaptive, potentially modifiable coping strategies arising in the context of family disruption come after the psychological screen to be labeled as an indicator of mental disorder. This, in turn, may inappropriately interfere with the child's adoption prospects. It is suggested that psychological screens for normalcy of preadoptive children represent a misuse both of psychology and psychiatry for they are motivated more by the needs and interests of social institutions involved in the adoption process rather than those of the child. PMID- 15810177 TI - Federal office on protecting human subjects issues new decision charts. PMID- 15810178 TI - Protection of children and risks with medical devices. PMID- 15810179 TI - Angry father accuses researcher of violating his family's privacy. PMID- 15810180 TI - Grant proposal reviewer says he would have scored proposal lower due to conflict of interest. PMID- 15810181 TI - New human subject protection regulations are being delayed. PMID- 15810182 TI - Protecting against financial conflict of interest at the NIH. PMID- 15810183 TI - IRB opts for "user friendly" letter rather than an informed consent form. PMID- 15810184 TI - Researcher said his colleague should have revealed possible conflicts of interest. PMID- 15810185 TI - In defense of an error--intellectual corruption in contemporary science. PMID- 15810186 TI - Should all patients who attempt suicide be treated? A response to Savulescu. PMID- 15810187 TI - The patient and the public good. PMID- 15810188 TI - Human ethics review and social sciences: several emerging ethical issues. AB - The extension of ethical review to research in the social sciences and humanities has raised a number of new issues for Institutional Ethics Committees. The emergent issues discussed here are: the relationship between subject and researcher, the use of data to which employment gives access, the impact of questionnaires, the impact of sampling techniques, the impact of observation, when the subject is an organisation, and conflicting rights to research. These issues underscore the importance of IECs maintaining a flexible and watchful oversight of research and the impossibility of a simple clutch of rules to guide ethical research in this domain. PMID- 15810189 TI - The Nancy Crick case. PMID- 15810190 TI - Nancy Crick, assistance to die and palliative care. PMID- 15810191 TI - Some legal reflections on the death of Nancy Crick. PMID- 15810192 TI - Fatherless families: how important is genetic relatedness? AB - How should families be constructed? Does it matter if we choose to ignore 'blood ties' and raise children without their genetic parents? The debate over a recent court ruling allowing single and lesbian women access to assisted reproductive technologies (ART's) illustrates two possible answers to this question. Many of those opposed to the ruling argue that the traditional biological family is the natural family unit and the ideal family form, which should be preserved. Amongst those in favour it is argued that families are not defined by nature but culturally constructed, that the traditional biological family is just one possible family form and that the way families are arranged is a matter of personal preference. This paper argues that questions about the importance of preserving the biological family rest largely on the value we attribute to genetic relatedness and not on whether the family is a natural or cultural construct. I argue against the claim that genetic relatedness and biological families are so important to children's welfare that lesbian and single women should be denied access to ART's. PMID- 15810193 TI - Is the Australian HREC system sustainable? AB - In Australia, Human Research Ethics Committees (HRECs) have a vital role to play- as the primary institutional mechanism for ethical review of research--in protecting research participants, and promoting ethical research. Their ability to act effectively in this role is currently threatened by the limited support they receive and their burgeoning workloads. In this discussion paper, I trace some of the factors contributing to what I describe as a resource crisis in human research ethics. I suggest a review of the working of HRECs to canvas a range of alternatives which might serve to redress this crisis, so as to ensure the continued effectiveness of HRECs in protecting participants and promoting ethical research. PMID- 15810194 TI - The expectation gap, risk management and the Australian HREC system. AB - The Australian HREC system is experiencing increasing workloads and greater public scrutiny. Dr Dodds asks whether the system is sustainable and aims to encourage a constructive critical debate about the system. This article suggests there is a gap between the demands on the system and expectations of researchers, regulators and the community. The evolution of the HREC system reached a significant milestone of the publication of the National Statement on Ethical Conduct in Research Involving Humans. This article argues that there will be increases in HREC commercialisation of research and clinical trials. The Andrews Committee has recommended another independent review of the system. The system is also under review by the joint ALRC/AHEC inquiry into the protection of human genetic information. PMID- 15810195 TI - Improving Australia's ethical review processes--slow and steady wins the race. AB - In this response, the Australian Health Ethics Committee (AHEC) indicates that it shares, and has strategies in place to address, the majority of the concerns identified by Susan Dodds. AHEC believes it is too early to assess the full impact of the National Statement on Ethical Conduct in Research Involving Humans (1999) or to call for a major review of the ethics committee process. While some Human Research Ethics Committees (HRECs) are overstretched, the system is not on the verge of collapse. PMID- 15810196 TI - Is the Australian HREC system unsustainable? AB - The Australian HREC system is now highly centralised and subject to national guidelines in an attempt to assure consistency of decision-making and of ethical standards. The penalty has been greatly increased paperwork and reporting requirements which many committees feel burdensome. In addition, many Committees may be under-resourced. However, I argue that though it is timely to reassess the current directions it is an exaggeration to claim that the system is in danger of collapse. PMID- 15810197 TI - Is the Australian HREC system sustainable--a rural perspective. AB - The scenario put forth by Dodds accurately reflects the current situation of the Australian HREC system and is difficult to take issue with. Those involved in providing input to HRECs continually confront the issues identified by the author. The increasing responsibility, the need for greater documentation, the increase in the range and complexity of applications, plus the increased ongoing surveillance of projects is placing an increased workload on HRECs. There are also some additional issues of relevance to HRECs operating in a rural environment. PMID- 15810198 TI - Response to Susan Dodds: is the Australian HREC system sustainable? PMID- 15810199 TI - Research ethics review and the bureaucracy. AB - This paper suggests that the increasing bureaucracy of ethics review by committee is more about fulfilling institutional requirements than it is about ethics. It is suggested that ethics committees should not be instruments of bureaucratic regulation and control. They should be freed to play a critical role within the institution, to support and develop ethical research and researchers, and given time to discuss and explore the difficult ethical issues where they arise. To burden research ethics committees with trivial bureaucratic tasks is to miss an opportunity for healthy exchanges of views about ethics and to distort the nature and meaning of ethics. PMID- 15810200 TI - Is it time to abandon institutional research ethics committees? AB - Research on human beings has significantly increased in ethical and scientific complexity. Ethics review is at a fork in the road. Either we significantly increase the resources we provide to support institutional research ethics committees. Or we abandon the institutional base of human research ethics review and move to model of expert suprainstitutional ethics committees. PMID- 15810201 TI - Susan Dodds' reply. PMID- 15810202 TI - The ethics of pharmacogenomics. AB - Of the future technologies arising from the Human Genome Project, pharmacogenomics will probably be the first to have a widespread impact on the everyday practice of medicine. This technology offers great benefits but also presents some difficult ethical challenges. This paper explains what pharmacogenomics is and examines three of the issues that it raises: orphan populations, the use of ethnicity in drug trials, and potential obstacles to informed consent for genetic testing. PMID- 15810203 TI - Public health--virtue ethics versus communitarianism: a response to Wendy Rogers. PMID- 15810204 TI - Open and closed committees. PMID- 15810205 TI - Doing retrospective child sexual abuse research safely and ethically with women: is it possible? Two perspectives. PMID- 15810207 TI - [The study of resonant multi-photon ionization mechanism in the vapour of sodium]. AB - By two-photon resonant three-photon ionization and two-photon hybrid resonant three-photon ionization in Na2-Na the different ion current strength with different temperature was measured. The result was discussed. PMID- 15810206 TI - Refusal clause seen as threat to reproductive health, gag on information. PMID- 15810208 TI - [Luminescence properties and energy transfer between Ce3+ and Tb3+ ions in KA1F4: Ce, Tb]. AB - Matrix KA1F4 was prepared through reaction of mixtures of solution K2CO3 upon certain solution of Al(OH)3 plus HF(40%),dehydrating at 100 degrees C and annealing for 48hr at 450 degrees C. A series of KA1F4 : Ce,Tb phosphors were synthesized by a solid reaction method. Their excitation and emission spectra have been investigated. It was observed that Ce3+ can effectively sensitize Tb3+ in KA1F4, so as to enhance the emission of Tb3+ significantly. KA1F4 :Ce,Tb might be a new high efficient green emitting phosphor. PMID- 15810209 TI - [Medium effects on derivative reaction between fluorescamine and primary arlyamine]. AB - The effects of pH, various organic solvents, surfactants, beta-CD and PVA on derivative reaction and fluorescence properties of its product between fluorescamine and p-amino phenyl sulfonic acid have been studied. The results show that the derivative reaction needs the organic solvent, many kind of organic solvents are available, but its amount should not be too large. The rate of derivative reaction was slowed and fluorescence intensity of product was decresed with increase in amount of organic solvent and precision is not good because of the fluorescence intensity is changing with delay time. Only tritonX-100 enhanced the fluorescence of product in all of the surfactants tested. Span20, Tween80 and Brij35 which same as tritonX-100 are non-ionic surfactant, and PVA, beta-CD do not exhibit sensitization. Ionic Surfactant CTAB, SDS quench the fluorescence. The sensitization of tritonX-100 is dependent on the amount of organic solvent. A competitive effect between tritonX-100 micellae and organic solvent is exhibition. In this paper, the derivative reaction between fluorescamine and p chloroaniline is compared and studied too. PMID- 15810210 TI - [Synthesis and spectroscopic properties of rare earth and p-anisic acid with 1, 10-phenanthroline]. AB - The title complex with the formula EuL3 x phen x H2O (phen=1, 10-phenathroline, HL=p-anisic acid) were synthesized and characterized by elemental analysis, molar conductivity, IR, UV, NMR etc. PMID- 15810211 TI - [Fluorescence lifetimes of squamous carcinoma, adenocarcinoma and normal human lung tissues in vitro by time-resolved fluorescence]. AB - Using pulsed dye laser 632.8nm, lifetimes of adenocarcinoma, squamous carcinoma and normal human lung tissues in vitro have been measured by time-resolved fluorescence as 5.1, 5.6 and 6.3 ns respectively. Time-resolved fluorescence shows that the decay time of tumor tissues is faster than that of normal tissues. The results may offer new diagnostic method to determine cancer. PMID- 15810212 TI - [The potential energy functions of hindered rotation for HF in rare gas lattices]. AB - The potential energy function of the hindered rotation of a linear molecule in a face-centered-cubic lattice is determined and that of HF in the rare gas lattices is also determined using the spectroscopy data. PMID- 15810213 TI - [The influence of KCl on SERS of fuchsin basic in silver colloids]. AB - Surface enhanced Raman scattering (SERS) spectra of fuchsin basic (FB) in silver colloids were measured. The influence of KCl on the SERS of FB was studied systematically with different excitation wavelengths (476.5, 488.0, 514.5, 632.8nm). The dependence of the relative SERS intensity of the 1366cm(-1) and 242cm(-1) bands of FB on KCl mole value at different excitation wavelengths were given. The SERS spectra of FB at 514.5nm and 632.8nm wavelength excitations were very different, and the relative intensity of 242cm(-1) to 1366cm(-1) SERS spectra was changed. PMID- 15810214 TI - [FT-IR characterization of extracted organic phase containing D2EHPA-lanthanide complexes and effects of adding DMHPA]. AB - FT-IR spectroscopy was used to characterize the extracted organic phase of di (2 ethylhexyl) phosphoric acid-n-heptane containing lanthanide ions Ln3+ (Ln: Y, Pr, Nd, Eu ). It was found that the coordination interaction of heavy lanthanide ions with D2EHPA is stronger than that of light lanthanide ions, and saponification percentage of D2EHPA affects both the absorption frequency and the intensity of P=O bond in the extracted organic phase. Furthermore, the addition of di (1 methylheptyl) phosphoric acid to the D2EHPA extraction system leads to the variation of both peak position and intensity of P=O bond. PMID- 15810215 TI - [Study on the photo-properties of 2-(2-substituted phenyl propenyl)-5-substituted benzoxazoles]. AB - The UV spectra and fluorescence emission spectra of the twenty title compounds were examined, and their substitution effect was discussed. The solvation effects of sixteen solvents on two compounds (compound 4 and compound 10) and the photo reaction of one compound (compound 9) were also studied. PMID- 15810216 TI - [IR study on the thermal decomposition of trinuclear and tetranuclear iron benzoate complexes]. AB - The title complexes shown as [Fe3O(C6H5COO)6(H2O)]Cl x 1.5H2O and [Fe4O2 (C6H5COO)7(bipy)2]ClO4 x 3.5H2O were prepared. Their thermal decomposition process was studied qualitatively and quantitatively by means of thermogravimetric analysis and infrared spectroscopic measurement of samples after heating at different temperatures. From the experimental results, the possible thermal decomposition process of the two compounds can be deduced. Additionally, according to the fact that the nu(as) (COO) of the two compounds decreases while the nu(s) (COO) remains unchanged as the heating temperature increases. It can be drawn that the carboxylate coordinative form possible change from bridged to chelate one. PMID- 15810217 TI - [Investigation of the carboxymethyl hydroxypropyl guar gum cross-linked with metal ions]. AB - In this paper the infrared-spectral changes and thermal gravimetric analysis of the gels formed with carboxymethyl hydroxypropyl guar gum (CMHPG) cross-linked with metal ions (Fe3+, Cr3+, Ti4+) have been studied. The results indicated that the coordinated active sites are mainly supplied with hydroxypropyl, neigbouring cis-OH of main chain and carboxymethyl groups of CMHPG. And the results of spectral analysis showed the strength orders and coordinated mode of the guar gum cross-linked with metal ions. PMID- 15810218 TI - [Study on evaluating quality of analgini powder pharmaceuticals by near infrared reflectance spectroscopy]. AB - The possibility of using near-infrared reflectance spectroscopy for the evaluating quality of analgini powder pharmaceuticals was investigated. We used the multivariate statistical classification technique to identify true, poor and false phamaceuticals of analgini from their first derivative spectra successfully. The results are satisfactory. PMID- 15810219 TI - [Spectroscopic investigation of the properties of axial coordination of tailed metalloporphyrins]. AB - The properties of axial coordination of nitrogenous ligands to four tailed metalloporphyrins, O-PyTPPFe ( III ) Cl, O-ImTPPMn ( III ) Cl, O-PyTPPMn ( III ) Cl and O-ImTPPFe ( III ) Cl, has been investigated by UV-Visible spectral techniques. The effects of terminal groups, solvents and metal center were examined and discussed. PMID- 15810220 TI - [Study on the Raman spectra of ZYA system]. AB - The Raman spectra of ZYA system [60wt% ZrO2 (2.25mol% Y2O3) -40wt% alpha-Al2O3] were measured before and after its pressing. The results proved the toughening mechanism of phase transformation from tetragonal ZrO2 to monoclinic. PMID- 15810221 TI - [Determination of Ni, Al, Mn and Si in copper based alloy by high current and microsecond pulsed glow discharge optical emission spectrometry]. AB - The determination of Ni, Al, Mn and Si in copper based alloy by high current and microsecond pulsed (HCMSP) glow discharge optical emission spectrometry (GD-OES) has been reported. With the laboratory built experiment set-up, the factors effecting analytical performance have been studied. The results of sample analysis are in good agreements with the recommendation values, and the stansard deviation for six determinations were 2.66% for Ni, 2.17% for Mn, 6.02% for Si, 1.00% for Al respectively. PMID- 15810222 TI - [Multi-elemental analysis by preconcentration on EDTA-cellulose microcolumn coupled to ICP-AES]. AB - EDTA-cellulose was prepared by chloroacetic acid covalently bound to filter flakes. Fe, Cu, Zn, V, Mn, Co, Ni, Al, Cd and Pb were preconcentrated on microcolumn filled with EDTA-cellulose and analyzed with ICP-AES after being eluated with dilute HNO3. At given pH value, alkali and alkaline earth metal ions can be quantitatively separated as matrix elements, and did not interfere eluate determine. High concentration factor was obtained. The usefulness of the method is shown by analyses of Standard Reference Materials. PMID- 15810223 TI - [Determination of Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES]. AB - A method of simultaneous and direct determination for Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES is reported. The spectral interferences and effect of acidity have been investigated. Working conditions were optimized. The method has been applied to the analysis of silver brazing filler metals with RSD of 4-7% and recovery of 94-105%. This method was accurate, simple and rapid. PMID- 15810224 TI - [Determination of calcium, magnesium and potassium in nurtured cell by AAS with quick-pulsed nebulization technique and NaOH base digestion]. AB - The sample is digested with 6% NaOH solution and an amount of 50 microl is used for protein content analysis by the method of Comassie Brilliant Blue G250, the residual is diluted with equal 0.4% Lathanurm-EDTA solution. Its Calcium magensium and potassium content are determined by AAS. With quick-pulsed nebulization technique. When a self-made micro-sampling device is used, 20microl of sample volume is needed and it is only the 1/10 approximately 1/20 of the sample volume required for conventional determination. Sensitivity, precision and rate of recovery agree well with those using regular wet ashing method. PMID- 15810225 TI - [Study on the direct determination of Pb and Cd in milk powder by GFAAS]. AB - The paper reports the direct determination of Pb and Cd in milk powder by GFAAS. The mothed does not need any pre-treatment to the samples and fully use some function of pre-treatment which the Graphite Furnace have. The result indicates that the method is simple, rapid and accurate, and the false result from any pre treatment to the samples can be reduced. The recover rate of Pb is 100.1%, CV =2.7 ; and Cd is 98.6%, CV = 3.1 %. This method is also applied to the rapid determination of Pb and Cd in soluble samples with satisfactory results. PMID- 15810226 TI - [Extraction-flame atom absorption determination of trace copper, zinc, cadmium and iron in bones]. AB - Trace content of Cu, Zn, Cd and Fe in bones is determined by flame atomic absorption spectrometry after preceding preconcentration using the extraction system O-phenanthroline-sodium perchlorate/1, 2-dichloroethane. Also, the factors of influence were studied. The method was simple, accurate and reliable, The detection limits were Cu: 0.038microg/ml, Zn: 0.0042microg/ml, Cd: 0.0019microg/ml, Fe: 0.02microg/ml. The recovery rate was between 92.8 and 105%. PMID- 15810227 TI - [Sequential determination of trace calcium, sodium, iron and zinc in import alumina by FAAS]. AB - This article introduces the sequential determination of the four elements by air acetylene FAAS. The method is based on using H3BO3-Li2CO3 as mixed melting agent, the alumina samples filled in platinum crucibes were melted at 1000 degrees C then extracted by HCl, and added the release agent, the protective agent and the eliminator of ionization, respectively, the four oxides contents were determined sequentially. The standard samples have been analysed many times by this method, the experimental results show that the determination errors are less than the tolerance. The values of test samples detected by present method are in accordance with those of by GB6609-86. This approach has been used for 100 and more lots of import alumina determination. It is not only accurate and reliable, but also easy and rapid. It is worth popularizing. PMID- 15810228 TI - [Determination of lead in cosmetic by graphite furnace atomic absorption spectrometry]. AB - A method is described for the determination of Pb in cosmetic by graphite furnace AAS. The samples were directly determined after most of them were solved. The dynamic range,detection limit, relative standard deviation and recovery are 0.020ng-1.6ng, 0.010ng, 0. 8-6.7%, 97-104% respectrively. PMID- 15810229 TI - [Determination of composition of electronic material by fundamental parameter method of XRF]. AB - The fundamental parameters (FP) method of XRF in determination of the composition of electronic ceramic materials is studied. The fundamental parameters needed in calculation are developed by math calculation and the program is developed in C language. The method can also be used in determination of the composition of alloy. Some samples are analysed by this method and the relative derivation is less than 3%. PMID- 15810230 TI - [Measurement and analysis of tumor tissue autofluorescence spectra]. AB - Measurement results of laser induced autofluorescence spectra of tumor tissue from little rat are given in this paper it is compared with normal tissue of the same body After analyzing the origin of tumor tissue autofluorescence and the difference with normal tissue spectra, it is concluded that autofluorescence spectra of tissue may reflect the features of tumor tissue and be utilized to diagnose the tumor. PMID- 15810231 TI - [Fluorescence spectra study on 2, 3, 7, 8-tetrachloro-n-ethyl-phenothiazine]. AB - In this paper we gave been discussed the variation law of fluorescence intensity of 2, 3, 7, 8-tetracloro-N-ethyl-phenothazine under different conditions. The experimental results show the best emit light system of fluorescence. It has theoretical significance and practical value on raising seneitivity of organic photoconductor. PMID- 15810232 TI - [Study of conditions on determination of benzo (a) pyrene by synchronous fluorimetry]. AB - The conditions of title method has been studied. In this paper coefficient correction was used to eliminate the interference of benzo(k)fluoranthene. The method has an advantage of covenient and rapid. The results of real samples are satisfactory. PMID- 15810233 TI - [Influence of extraction on fluorescence of the subrenite in the peat]. AB - Comparative study on the subrenite in the peat and extracted remain has been made. The study reveals that extraction has considerable influence on the fluorescence of the subrenite. Before extracted, the subrenite in the peat gives out violently fresh yellowish green to green fluorescent light ,and the lambda(max), ranges from 470 to 520nm and the Q650/500 from 0.10 to 0.65, but after extracted, the subrenite in the extrated remain gives out weakly dark yellow to orange-yellow fluorescent light and the lambda(max) ranges from 605 to 660nm and the Q650/500 from 1.85 to 4.09. The extraction has made the fluorescent intensity of the subrenite weak. PMID- 15810234 TI - [Extended Kalman filtering trained neural networks and multicomponent analysis of amino acids]. AB - The principle of neural networks was briefly described. Backpropagation (BP) is a widely used algorithm to train NN with slow convergence and local optima. In order to overcome the above weakness of BP, a novel learning algorithm, extanded Kalman filtering (EKF, EF), was proposed with high performance. EFNN was used for simultaneous multi-component analysis with good results and no separation. PMID- 15810235 TI - [A laplace transform method for analysis of fluorescence dynamics]. AB - A Laplace transform formula is developed for analysis of a multi-exponential fluorescence dynamic process. A dynamic spectrum function is introduced, calculation of which directly yields parameters of the corresponding dynamic process. The formula is applied to analyze fluorescence dynamics of beta subunit from R-phycoerythrin. It is demonstrated that electronic excited-states of the biomolecular system exhibit two exponential decay processes, the faster of which relates to excitation equilibrium among chromophores, and the slower to radiative relaxation of the chromophores. PMID- 15810236 TI - [The Raman scatting of carbon nanotubes]. AB - Raman scattering measurements of the cathode deposite produced by carbon arc discharge method and graphite was repotred. The results indicate that the inner core of the deposite containing large quantities of carbon nanotubes undergoing crystallization process. Carbon nanotube is one kind of micro-crystal of quasi graphite. PMID- 15810237 TI - [Simple formulae for the calculation of Li-like and Be-like ionic energies of ground states]. AB - Using the two-parameter variational method, the simple formulae for the calculation of Li-like and Be-like ionic energies of ground states has been derived, which show convenience in calculation and accuracy. PMID- 15810238 TI - [SERS and electrochemistry studies on the inhibition of thiourea and phenyl thiourea on iron]. AB - An iron electrode was obtained by depositing a thin layer of iron on active silver electrode with overpotential deposition method. The E - I curve and SERS spectra of thiourea (TU) and phenyl-thiourea (PTU) adsorbed on the iron electrode surface were studied by means of electrochemistry and SERS spectroscopy. The results showed that both TU and PTU can be adsorbed on the iron electrode surface and the inhibition of PTU is better than TU in 0.1mol/L KCl solution. The adsorption mode of TU and PTU is similar in the interaction of S atom and iron surface. The cause of better inhibition effect of PTU lies in that the PTU can be more easily adsorbed on iron electrode because of more negative charge in its S atom and benzene ring in PTU is arranged away from the iron surface to form a rather compact adsorption layer due to steric hindrance. PMID- 15810240 TI - [Investigation of fluorescence spectra of alum floc]. AB - The sharp fluorescence peak of alum floc was observed at 340nm. The peak height at 340nm is in proportion to alum concentration under same degree of neutralization. There is similar regular change of this peak height as change of neutralization degree under different alum concentration. PMID- 15810239 TI - [Study on synthesis and molecular spectra of the N-(2-hydroxy) naphthylmethyleneamino iron(III), cobalt(II) and copper(II) complexes]. AB - The title complexes were synthesized via reaction of iron nitrate, cobalt acetate and copper acetate with the didentate O,N-donor Schiff bases in methanol solution. The electronic and infrared spectra of these complexes were measured and the major IR bands were assigned. PMID- 15810241 TI - [Study on the effect of cyclic adenosine monophosphate on human erythrocyte membrane by FTIR and computer aided analysis]. AB - The effect of cAMP on erythrocyte membrane proteins was studied by FITR, deconvolution and curvefitting. It was found that the change of cAMP's concentration affects the secondary structure of membrane proteins. The regulation of cAMP has a best concentration region, during which cAMP has the strongest effect. PMID- 15810242 TI - [Determination of alkali-earth metal by microwave plasma torch atomic emission spectrometry]. AB - Determination of alkali-earth metal by microwave plasma torch atomic emission spectrometry (MPT-AES) was investigated. Operating conditions the effects of some coexistent elements were examined. Detection limits for Be, Mg, Ca, Sr and Ba are 0.47, 1.4, 0.15, 0.20 and 2.6ng/mL, respectively. Method has been applied to analyse some actual samples with satisfactory results. PMID- 15810243 TI - [Enhancement effect of spectroscopic carriers on emission intensities in laser source plasma]. AB - Enhancement effect of easily volatile elements I (boiling point = 184.35 degrees C) and S (boiling point = 444.67 degrees C) on spectral line intensities of various trace elements in laser source plasma were investigated by experiments. In order to discuss mechanism of enhancement effect, some of the plasma parameters (Texc,ne) were measured. PMID- 15810244 TI - [Spectroscopic diagnostic of atmospheric pressure argon microwave induced plasma]. AB - Atmospheric argon microwave-induced plasma (Ar-MIP) sustained by a surfatron was obtained. Emission spectra and characteristics of argon metastable (1Sj) were studied with an optical multichannel analyer (OMA-3, 1460) under different experimental condition. Excitation mechanism of the plasma was discussed. PMID- 15810245 TI - [Determination of trace phosphor in aluminum alloy by AES using background correction with spark source]. AB - Using background correction with spark source by AES, fixed background correction, successfully corrected the dynamic background of phosphor, results obtained for determination of phosphor in aluminum alloy are satisfactory. PMID- 15810246 TI - [Determination of molybdenum in natural waters by direct current plasma atomic emission spectrometry with ion-exchange preconcentration]. AB - A new method was developed for the determination of trace molybdenum in natural waters by direct current plasma atomic emission spectrometry after separating and preconcentrating with D301 anion exchange resin. Mo adsorbed on the resin column can be recovered quantitatively by using 20mL of 8mol/L HNO3 at a flow rate of 0.6mL/min with elution rate of (81.6 +/- 1.78)%. The behaviour of Mo on the resin has been discussed. Simulative water and natural water samples have been determined. It was found that the interference element Ca could be separated effectively from Mo; an enrichment factor of 50 was obtained for Mo in 1L water sample; the determination limit of Mo was improved to 0.5 microg/L and the recovery of 90-107% was obtained. PMID- 15810247 TI - [Determination of impurities in trimethylgallium by ICP-AES]. AB - In this article, it was reported that ICP-AES was used to analyze impurities in trimethylgallium. Gallium matrix could not produce background and spectral overlapping interferences. PMID- 15810248 TI - [Direct simultaneous determination of o, m, p-nitronethylbenzene in mixture using spectrophotometry combined with Kalman filter]. AB - The basic principle of Kalman filter and its modified method, including forgetting factor and enhancing dimensions of vector, for the error correction of measurement model in spectrophotometric determination of multicomponents were introduced. Combining forgetting factor with enhancing dimensions of vector, the recursive effect of Kalman filter method was improved. The result of modified Kalman filter was compared with other three chemometric methods, including target transformation factor, iterative target transformation factor and artificial neural network. This method can apply to direct simultaneous determination of o, m, p-nitronethylbenzene in mixture without any preliminary chemical separation. PMID- 15810250 TI - [Characteristic mass and atomization efficiency of graphite furnace atomic absorption spectrometry]. AB - Measurements of peak height and peak area characteristic masses and atomization efficiency for 7 elements (Ag, Bi, Cd, Cr, Ga, Ge and In) have been studied. The m0(i)(exp) values are compared with theoretical data, m0(i)(cal), to assess the effect of temperature on the correction factor of theorization for characteristic mass value (alpha = m0(i)(cal) m0(i)(exp)), the peak height and the peak area atomization efficiencies (betap and betai). The peak area atomization efficiency (betai) was equal to the correction factor of theorization for characteristic mass value (alpha) times tauD/tauR value. The tauR was value experimentally measured of atomic residence time and the tauD value calculated from rD = l2/8D. The betai value of elements always are smaller than the alpha values because the tauD/tauR values is less than one for most elements in the operating conditions. Past of the analyte vapours is readsorbed by the heated tube wall at high temperature and reevaporation of the adsorbed analyte followed by diffusion account for the larger atomic residence time and it appeared that the tauR value was larger than the tauD in graphite furnace. PMID- 15810249 TI - [Research of the effect of ascorbic acid prereduced Pd in the determination of micro quantity of Mn with GFAAS]. AB - In this article, method of determination of micro quantity of Mn with GFAAS and Pd have been researched, results of different additional methods of Pd were compared the combining energy of Pd in three heating steps were determined by X ray Photoelectron Spectroscopy. The point was given that as a catalyst reduced Pd changed courses of Mn atomization, and when pre-reduced Pd was used because of its much larger surface area, method had much better RSD and had much stronger signal than other methods in those cases Pd was used too. PMID- 15810251 TI - [Simultaneous determination of cobalt and indium by atomic absorption spectrometry]. AB - Present paper describes method which uses solution of simultaneous equations to compensate for spectral "overlap" interference of In 252.137nm with Co 252.136nm. Experimental results show that solution of simultaneous equations is possible method for resolving spectral interference in atomic absorption spectrometry. This method has been applied to determination of Co and In in the samples. Satisfactory results are obtained. PMID- 15810252 TI - [Determianation of trace copper, iron and lead in purified AgNO3 with liquid membrane enrichment]. AB - In this paper a new method of preconcentrating microimpurity Cu, Fe and Pb in high pure AgNO3 using emulsion liquid membrane technique was developed for FAAS determination. The enrichment multiple was 25, recoveries were over 95%. Matrix silver does not interfere enrichment and determination. PMID- 15810253 TI - [Determination of Cu, Pb, Sn, Fe and Zn in the ancient Chinese bronzes by X-ray fluorescence filter paper method]. AB - The determination of Cu, Pb, Sn, Fe and Zn in the ancient chinese bronzes has been performed using X-ray fluorescence filter paper method. The result is satisfactory. The samples are dissolved in the HCl-H2O2 system, ard then the solution is dropped on the filter paper. It is convenient for detection by XRF. In addition this method is efficient to reduce the matrix effect of sample. The correlative coefficients of calibration curves between element concentration and X-ray intensity are over 0.99. The rate of recovery is 97-104%, and the relative standard deviation for each element is less than 2.00%. The copper's result measured by this method agrees well with that of chemical method. This method is simple, rapid and accurate. It is also applied to determine the elements in the other ancient objects. PMID- 15810254 TI - [Application of wavelet transform to the isolation of EXAFS oscilations from experiment data]. AB - It is of significant importance to isolate lambda(k) form the total absorption cofficient data micro(E) for the analysis of EXAFS spectrum. Generally, least squares procedure of polynomial spline or B spline was used for removing the smooth sbsorption background, which has been proved to be effective. But it suffers in that the trial of knot points is time-consuming and it is poor in reproducibility. In this paper, a new chemometrics method--wavelet transform analysis was used to separate the lambda(k) and background absorption from micro(E) and it is proved that the technique is a very good method for the analysis of EXAFS spectrum. PMID- 15810255 TI - [Analysis of coating on painted pottery of new stone age of Xi-shan ruin and relation with sintering temperature]. AB - X-ray diffractoermeter and X-ray fluorescence methods have been used to analyse coating on painted pottery of new stone age of Xi-shan ruins. According to experimental results, sintering temperature of ancient pottery is inferred. PMID- 15810256 TI - [The method for studying kinetics of formation reaction of complex in dimeric system]. AB - The method for studying kinetics of formation reaction of complex in aggregation system was reported in this paper. The concentration of more than one species was calculated by improved method for calculating dimeration constants. Kinetics of reaction between Cu (II) and tri-sulfophthalocyanine (S3Pc) was studied in this way. It shows that the rate of reaction between Cu(II) and S3Pc can be expressed as follows: v = K[Cu][S3Pc] PMID- 15810257 TI - [The aggregation and kinetics of decomposition reaction of Ga sulfophthalocyanine]. AB - The aggregation and kinetics of decomposition reaction of Ga-sulfophthalocyanine has been studied by electronic spectroscopy. The effects of pH on the aggregation are discussed. The experimental results show that the rate of decomposition reaction of Ga-sulfophthalocyanine can be expressed as follow: -dcM/dt = 12.279cDcOH-. Where cD and cOH- concentration of GA-sulfophthalocyanine and OH- respectively. PMID- 15810258 TI - [Study on determination of composition of complex by multiwavelength R factor method]. AB - The pure spectrum of the complex is obtained by R factor method to eliminate the absorbance of equilibrium concentration of residual reagent. The reagent concentration is determined by multiple linear regression method. As initial concentrations of metal and reagent are changed, and the stable constant is not changed, complex composition and its constant are acquired by means of trial method, in which the input parameters, m and n, are selected. Then the K value is calculated and the minimum value of the total derivation in K is found. By this method, the composition of complex in systems such as Pb-XO, Al-CAS and Zn-PAR are all to be 1:2. PMID- 15810259 TI - [Spectrophotometric studies on and applications of the reaction of copper with 2 hydroxy-3-carboxy-5-sulfobenzenediazoaminoazobenzene]. AB - A highly sensitive color reaction of copper(II) with 2-hydroxy-3-carboxy-5 sulfobenzenediazoaminoazobenzene (HCSDAA) in the presence of Triton N-101 was studied in detail. In borax buffer medium at pH 10.4-11.5, the red complex formed has a maximum absorption at 530 nm and its apparent molar absorptivity is 1.31 x 10(5)L x mol(-1) x cm(-1) and sandell sensitivity is 4.85 x 10(-4) microg x cm( 2). The molar ratio of copper(II) to HCSDAA in the complex is found to be 1:2. Beer's law is obeyed in the concentraction range of 0-0.24 microg/mL for copper. The proposed method has been applied to the spectrophotometric determination of microamounts of copper in ore and peanut with the relative error and the relative standard deviation (n = 6) of less than 3.7% and 1.6%, respectively. PMID- 15810260 TI - [Study of spectrometer calibration experiment research]. AB - Spectrometer calibration is very important for quantitative research of imaging spectral remote sensing. After spectral calibtation and radiometric calibration, it was known that the average shift of wavelength of SE590 channels was 8nm, and the linear response was depended on the change of energy recieved by SE590, and spectral response resolution was unreasonable beyond 950nm range. At last, the calibration results were proved by the field experiment measurement data. PMID- 15810261 TI - [Application of separation technique of chelation-adsortion in separation and enrich of microelements]. AB - This paper reported a new technique of the separation and enrich of microelements in spectra analysis. PMID- 15810262 TI - Basic data on arthritis knee, hip, and sacroiliac joints in adults ages 25-74 years. PMID- 15810263 TI - [Three-dimensional volumetric display--a new application of frequency up conversion]. AB - This paper discusses the present research situation and the application prospect of 3-D volumetric display technology. The possible usability and science significance of frequency up-conversion in 3-D volumetric display field are also demonstrated systematically. In ZBLAN: Pr, Yb glasses, we primarily demonstrated and achieved a new application of frequency up-conversion three dimensional volumetric display, using the two-frequency up-conversion of the rare earth ions. The efficiency of luminescence of Pr was enhanced much by the energy transfer between Pr and Yb ions. PMID- 15810264 TI - [XPS and MS study of phosphorous heterocycle compounds]. AB - 7 new phosphorous heterocycle compounds have been studied by XPS and MS. From XPS and MS data we have discussed the existence form and manner of the compounds isomer, and verified that the isomer only exists in gaseous state and liquid state. PMID- 15810265 TI - [Study of soft X-ray radiation in 8-20 nm region from a laser plasma source]. AB - A repetitive-laser-produced-plasma (LPP) soft X-ray source with a pulse repetition rate up to 10 Hz was developed. Spectra ranging from 8 to 20 nm were obtained from plasmas generated respectively by a 100 mJ (lambda = 0.53 microm) and 800 mJ (lambda = 1.06 microm) Nd:YAG laser focused on the following different target materials: C, Mg, Al, Ti, Fe, Cu, Zn, Sn, W, Pb and Bi. The dependence of spectral distribution on the target materials and laser parameters, such as pulsed laser energy, laser focus power density etc. are analyzed. The intensities of the soft X-ray radiation with different atomic numbers are characterized. PMID- 15810266 TI - [Preliminary study on radiant spectra of the leaves of paddy and upland rice]. AB - The characteristics of the radiant spectra (i.e. absorption, reflectance and transmission) of leaves of paddy and upland rice were studied. The experiment proves that the characteristics of the radiant spectra of the seedling leaves of paddy and upland rice is basically the same in the range of 200-800nm, and their chlorophyll content of the leaves is also quite near (the average values for paddy and upland rice are 4.25mg/g and 4.28mg/g, respectively). However, the absorption of the flag leaves of upland rice is higher and the transmission is lower compared to paddy rice in the heading-flowering period, while the reflectance is almost the same except for 550nm. The chlorophyll content in the leaves of upland rice is approximate twice as that of paddy rice (the average values of paddy and upland rice are 0.86mg/g and 1.86mg/g, respectively). PMID- 15810267 TI - [The calculation of transitions 4s(2)4p-4s4p(2), 4s(2)4p-4s(2)5s, 4s(2)4p 4s(2)4d, 4s4p(2)-4p(3) and 4p(3)-4s(2)5s energy levels, wavelengths and oscillator strengthes in Ga-like ions from Z X-Rh XV]. AB - In this paper, the 4s(2)4p, 4s(2)4d, 4s4p(2), 4p(3) and 4s(2)5s configuration's energy level structures, and configuration interactions in Ga-Like from Ga I-Xe XXIV ions are analysed in theory. The law of isoelectronic series is given. By using our fitted formula, the energies of Tc XIII ion 4s(2)4p, 4s4p(2) and 4s(2)5s configurations are predicted by interpolation, the energies of Tc IIIX-Rh XV ions 4p(3) and 4s(2)4d configurations are predicted by extrapolation. Wavelengths and oscillator strengthes (HXR theoretical calculation) of transitions 4s(2)4p-4s(2)4d, 4s(2)4p-4s4p(2), 4s(2)4p-4s(2)5s, 4p(3)-4s(2)5s and 4s(2)4p-4p(3) are given. The deviations of predictive energy levels from ob served values are less than 100cm(-1). PMID- 15810268 TI - [The vibration-rotational analysis of the high resolution ir spectrum of the CH3D molecule]. AB - High-resolution Fourier transform spectra of the band of the CH3D molecule near 6428cm(-1) have been recorded on a BRUKER IFS 120HR Fourier-transform spectrometer with multipass cell with a length of 20m, an unapodized resolution of 0.004cm(-1) was achieved. Vibration-rotation analysis in Local-mode can provide accurate enough initial values to predict the band origins. The vibration rotation transitions of CH3D molecule were assigned by using the ground state combination differences and least-squares fitted to yield ten vibration-rotation constants for the band of CH3D molecule near 6428cm(-1). Vibration assignment in Normal-mode and Local-mode expressed that the band of CH3D molecule near 6428cm( 1) is 3V2 vibration states of CH3D molecule. PMID- 15810269 TI - [Study on the NIR-Raman spectra and NIR-SERS spectra of amsacrine]. AB - The near infrared surface-enhanced Raman spectra (NIR-SERS) of amsacrine,an anticancer agent, on silver collids were recored with high quality. The NIR-SERS enhancement factor is up to 10(4) magnitude. In comparison with the near infrared Raman (NIR-Raman) spectra of solid amsacrine, more information of vibrational structures of amsacrine molecules at low concentration and almost in its practical application envi ronment, can be given by the NIR-SERS. The enhancement mechanism is associated with the adsorption of acridyl ring and sulfonyl on silver surface. PMID- 15810270 TI - [FTIR-PAS of rare earth coordinated compounds of Schiff base derived from vanillin and p-toluidine]. AB - The Fourier transform infrared photoacoustic spectra (FTIR-PAS) of Schiff base derived from vanillin (3-methoxy-4-hydroxybenzaldehyde) and p-toluidine and its rare earth coordinated compounds in the 3800-200cm(-1) were measured and inverstigated. The coordinated compounds have two classes: [LnL2Cl x 3H2O]Cl2(Ln:La,Ce,Pr,Nd,Sm,Eu,Gd,Dy;L:Schiff base ligand) and [LnL2Cl2 x 2H2O] Cl x H2O(Ln:Ho,Er,Tm,Yb,Lu). In the coordinated compounds there is intermolecular hydrogen bond derived from phenolic hydroxy group. The bands at 400 and 300cm(-1) are respectively assigned to bonds M-O (water) and M-N. PMID- 15810271 TI - [Studies on infrared spectra of Schiff base ligands and Mn(III) complexes with methylene bridge]. AB - The FTIR spectra (3800-150cm(-1)) of Schiff base ligands and Mn(III) complexes with methylene bridge were measured and investigated. The major bands have been empirically assigned in comparison with a large number of previous results. The characteristic absorption bands of intra-molecular hydrogen bond of ligands were found at ca. 3438cm(-1). The vC=N of ligands and complexes were found at ca. 1630cm(-1). The characteristic absorption bands of the phenyl ring of ligands and complexes were found at ca. 1630cm(-1). The characteristic absorptin bands of the phenyl ring of ligands and complexes were found at ac. 1598, 1469 and 755cm(-1). The vibrations coupling between the M-N stretching modes and the phenyl ring deformation were found at ca. 369cm(-1). The vibrations coupling between the M-N deformation modes and the phenyl ring deformation modes were found at ca. 247cm( 1). The vibrations coupling between the M-O stretching modes and the phenyl ring deformation were found at ca. 329cm(-1). The vibrations coupling between the M-N deformation modes and the phenyl ring deformation modes were found at ca. 275cm( 1). The vMn-Cl of the complexes were found at ca. 303cm(-1). The vMn-N of the complexes shift to higher wavenumbers and the vMn-Cl shift to lower wavenumbers than of the phenylporphyrin Mn(III) complexes. PMID- 15810272 TI - [Characteristic of vibration spectra for hydration state of herring sperm DNA fibers]. AB - By different temperature treatment, the herring sperm DNA fibers at various relative humidity have been analyzed using Raman scattering spectroscopic method for estimation of bound water content in fiber according to the typical peak intensity ratio between deoxyribosyl C-H stretching vibration (at 2964cm(-1)) and bound water O-H stretching vibration (at 3204cm(-1)). We found the bound water content of the fibers to be 9-14 water molecules per nucleotide pair (i.e. base pair) at variance with the calculated value of calf thymus DNA fiber by Tao's Raman test formula, but in similarity to the determined value by IR spectroscopy, X-ray diffraction, BET gravimetric analysis and differential scanning calorimetry analysis. Thus, herring sperm DNA fibers should have the same hydration state as both calf thymus DNA and salmon sperm DNA, that has been confirmed with the new formula: 2.5926 x I2964/I3204. PMID- 15810273 TI - [FTIR study on structure of pearl and nacre of mollusk shell]. AB - Pearl and nacre of mollusk shell (Hyriopsis cumingii) were studied with high resolution Fourier transformation infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The results of X-ray diffraction indicate that the samples are still aragonite after thermal treatment at 160 degrees C for 5hr, however, the significant variations of peak position and band width of FTIR spectra reveal the interaction between organic matrix and inorganic crystal in biomineralized materials. The variations of spectra were studied by a curve-fitting algorithm and the band related to the interaction is revealed. PMID- 15810274 TI - [The influences of TBP on the partially saponified DMHPA-heptane systems and the extracted organic phases containing rare earth]. AB - The aggregation states of partially saponified [di-(1-methylheptyl) phosphoric acid] (DMHPA)-tributyl phosphate (TBP) heptane (C7H16) and the extracted organic phases containing rare earth were studied. The result demonstrates that TBP can widen the range of the microemulsion formation in DMHPA-C7H16-NaOH-H2O systems. It also influence the states of the extracted organic phases of DMHPA-RE (III) and the behavior of water clearly. The gel could be formed in DMHPA-TBP-RE (III) systems under proper conditions. FT-IR spectra analysis shows that both DMHPA and TBP may coordinate with RE3+ and the hydrogen bonds of TBP with DMHPA and H2O contribute to the three dimensional network structure of the gels. PMID- 15810275 TI - [Application of surface-enhanced Raman spectra to the analysis of Chinese Ephedra soup medicines]. AB - A new method was developed to analyse the spectra of ephedrine in Chinese ephedra soup medicines, using surface-enhanced technique to combine thin layer chromatographic (TLC) technique with surface-enhanced Raman spectroscopy (SERS). The study indicates that the main vibrant characteristic spec tral band of the ephedrine molecules can be obtained by TLC in the samples of about 8 microg, and expounds char acteristics of the sample molecules and the silica gel. Therefore, it is clarified that the main chemical composi tion of Chinese medicines can be carried as finger-print type appraisal by combining TLC and SERS. PMID- 15810276 TI - [Measurements of Ar(I) excitation temperatures and electron number densities in an ICP with and without the presence of Freon 12--the development of ICP technology for hazardous waste management]. AB - For the management of refractory hazardous wastes, an innovative technology emerging recently is the application of ultra high temperature plasma. The preliminary study on destruction of Freon 12 by ICP (1-2kW) under a joint program between Xiamen Univ. and Hong Kong Baptist Univ. showed that the ICP technology indeed holds a great potential for hazardous wastes management. The destruction efficiency is more than 99.9999%. With and without the presence of Freon 12, Ar(I) excitation temperatures were measured by Boltzmann plot method and electron number densities the by H (beta) line broadening method. It was found that above the load coil, the excitation temperatures and electron number densities decrease with increasing the amount of Freon 12 presented in the central channel of the ICP, but inside the load coil region, the excitation temperatures are affected little by Freon 12. The conclusion of thermal pinch could be expected from this phenomenon. PMID- 15810277 TI - [Recent development of studies on the interferences and their correction methods in ICP-AES(I)]. AB - The recent development of the research of the spectral interferences and non spectral interferences, and the application of correction methods for interferences in ICP-AES were reviewed. PMID- 15810278 TI - [An effective method of mutual interference coefficients for correcting spectral interferences in routine ICP-AES for multielement analysis]. AB - An effective method was used to correct spectral interferences in routine ICP-AES for multielement analysis. In this method, all the components in an analysed object were considered to be related to each other, a strict mathematical model was applied and the mutual interference coefficients obtained from the usual artificial samples were replaced by those from natural standard samples which had the some matrix as the samples to be analysed. These mutual interference coefficients were used to correct the analysis data of GSD samples, and the results were mostly more precise than those corrected by the coefficients obtained from artificial samples. The correcting speed of this method is quick enough to satisfy the requirement of real-time correcting of analysis. PMID- 15810279 TI - [Spectral interference correction for ICP-AES by factor analysis and a comparison with other methods]. AB - Factor analysis method was used to correct spectral interference in inductively coupled plasma atomic emission spectrometry (ICP-AES) in the present work. The spectral overlap and structural background interferences were investigated and discussed with two real examples. Results show that the factor analysis is comparable to partial least squares (PLS) and the Kalman filtering in correcting the interferences. PMID- 15810280 TI - [Application of simple OMA system for rapid spectrochemical quantitative analysis to steel sample composition]. AB - In this paper, the experimental method and characteristic of a simple optical multichannel analyzer (OMA) system used for rapid spectrochemical quantitative analysis was introduced. Mangangese and chromium contents in different steel samples were measured. The reliability of experimental results was discussed according to the mathematical statistics theory. Analytical results were compared with those of chemical and ICP analysis. PMID- 15810281 TI - [Determination of net analyte emission intensity and signal-background ratio of Mg, Cr, Cd and Mn in Ar-air mixed coolant gas ICP]. AB - The dependence of net analyte emission intensity and signal-background ratio on coolant gas composition and observation heights for 8 lines of four elements (Mg, Cr, Cd and Mn) in Ar-air mixed coolant gas ICP was measured under the similar condition as that in the low power Ar-ICP with a commercial sequential plasma spectrometer. Further, the detection limits under compromise conditions were estimated. The results show that ion lines and neutral atom lines with high excitation energy have maximum intensity and are more intense than those in the Ar-ICP, when 5-10% air is added to the coolant flow. The intensity of neutral atom lines with low excitation energy drops gradually as air is added to coolant flow. The signal-background ratio of analyte lines depends upon excitation energy and wavelength. PMID- 15810282 TI - [Determination of trace Pb, Cu and Cd in environmental sample by slotted-tube atom-trapping atomic absorption spectrometry]. AB - The optimal measuring conditions of slotted-tube atom-trapping atomic absorption spectrometry of Pb, Cu and Cd were examined in detail. The results show that the characteristic concentrations measured after trapping for 1min under definite conditions were 1.82 x 10(-3), 1.26 x 10(-3) and 6.7 x 10(-5) mg/L for Pb, Cu and Cd, respectively. Thus slotted tube atom-trapping AAS yields increase in sensitivety for Pb, Cu and Cd by 110, 39 and 150 fold, respectively, in comparison with the conventional FAAS. Trace elements Pb, Cu and Cd in earth surface water, tea and hair were determined by slotted tube atom-trapping. Satisfactory results were obtained. PMID- 15810283 TI - [Method for determination research of trace germanium in the serum of cancer patients by graphite furnace atomic absorption spectrometry]. AB - Using a tungstate-coated graphite tube, trace germanium in serum samples was determined by Zeeman graphite AAS with NH4NO3 as a matrix modifier. The sensitivity of Ge can be significantly improved. The matrix interference has been removed. The relative standard deviation is less than 3.61%. The method is simple, rapid and accurate. PMID- 15810284 TI - [GFAAS determination of trace Cr(III) and Cr(VI) after its separation by coprecipitation-floatation with aluminium hydroxide]. AB - A method for determination of trace Cr(III) in natural water and drinking water by GFAAS after separation by coprecipitation-floatation with Al(OH)3 has been studied. Cr(VI) was determined also after being reduced by FeSO4. Conditions of Cr(III) floatation such as pH, the amount of floatation agent and coprecipitator Aluminium added, the flow rate of nitrogen gas, the floatation time etc. were discussed. The influence of coexisting elements was investigated. The detection limit is 0.027 microg/L (n = 15). For Cr(III) concentration level of 0.5 microg/L, the RSD is 3.4% and the recovery of Cr(III) is 96.0-104.0%. PMID- 15810285 TI - [Research on the determination method of trace amount of Pb and Cd in animal bone]. AB - A method for the determination of trace amount of Pb and Cd in animal bone using GFAAS has been established. The samples were determined directly after being diluted by TMAH solution. The method is handy and fast. The recovery is 96.95 103.15%. The RSD is less than 4.81%. PMID- 15810286 TI - [Study on determination of poisonous heavy metal element lead by graphite furnace atomic absorption spectrometry]. AB - In this paper, the determination of trace lead in foods by optical temperature control GFAAS was studied. The method was certified by the analysis of the national standard reference materials. PMID- 15810287 TI - [A new approach to correction method for matrix effect in X-ray fluorescence spectroscopy analysis]. AB - With the help of fundamental parameter program developed by the authors, the matrix effect of multicomponent complex sample and the composition of each element's X-ray fluorescence intensity are calculated. A new mathematical model for correcting multicomponent matrix effect is deduced theoretically. This model provides a new method to calculate influence coefficients alpha and beta. The correction model and the influence coefficients introduced by this paper have clear and definite physical meaning, which give satisfactory result when utilized in analyzing stainless steel sample. The paper also indicates that the effect of tertiary fluorescence can not be neglected in some cases. From the result of comparison experiments, we draw a conclusion that the tertiary fluorescence effect must be considered while analyzing Cr in a steel sample with high concentration of Ni. PMID- 15810288 TI - [Fluorescence emission spectra of petroleumsulfonate and OP-10 in multi components aqueous solutions]. AB - The determination of surfactant(s) in the alkaline/surfactant/polymer flooding solutions was an uneasy job. For this purpose, the method of fluorescence emission by phenoxy-containing surfactants in dilute solutions is suggested. The fluorescence emission intensity changes linearly with in the extent of low surfactant concentrations below the critical micelle concentration (CMC) and is not influenced by the presence of alkali, polymer PHPAM and salts. Under certain conditions the fluorescence emission by petroleum sulfonate and OP-10 in a multi components aqueous solution can be detected independently. This method is high selective, sensitive (the lowest detectable concentration 10(-7) mol/L) and microanalytical (the amount of test solution 2-3 microL). PMID- 15810289 TI - [Study on the determination of tung oil adulterated in vegetable oils by first derivative spectrophotometry]. AB - A method of first derivative spectrophotometry for qualitative and quantitative analysis of tung oil adulterated in vegetable oils, including peanut oil, bean oil, rape seed oil, tea seed oil, palm oil and mixed vegetable oil, was established. The spectrum of tung oil features three valleys at 291.3, 278.3 and 266.4nm, and three peaks at 284.1, 271.5 and 260.7nm. At 291.3nm, the coefficient (deltaE(1%)1cm/deltalambda) was -1.03 x 10(3). When the concentration of tung oil adulterated in vegetable oils was downed to 0.1%, the above specialities still remain and the changes in wavelengths were not more than 0.7nm. The detection limit of the concentration of tung oil adulterated in vegetable oils was lower than 0.1%. PMID- 15810290 TI - [Studies on the liquid-liquid extraction behaviour of Co(II) and Ni(II) without organic solvents]. AB - In PEG-Eriochrome cyanine R-(NH4)2SO4 system the liquid-liquid extraction behaviour of Co(II) and Ni(II) without organic solvents was investigated. We found that Ni(II) was almost extracted by PEG phase while Co(II) was not extracted in the water solutions of NaAc-HAc (pH=5) and (NH4)2SO4. Thus Co(II) was quantitatively separated from mixed solutions of Ni(II) and Co(II) ions. PMID- 15810291 TI - [Study of photoreflectance in Cd1-xMnxTe/Cd1-yMnyTe superlattices of II - VI diluted magnetic semiconductors]. AB - The observation of photoreflectance spectra of Cd1-xMnxTe/Cd1-yMnyTe superlattices was carried out at temperature T = 80K. The samples were grown by molecular beam epitaxy (MBE) technique. The heavy-and light-hole excitonic transition structures 11H, 22H, 33H and 11L were observed and the theoretical calculations, including the strain effects, were performed. By comparison, both agree well. PMID- 15810292 TI - [A multiexponential model in cavity ring-down absorption spectroscopy]. AB - A multiexponential model of cavity ring-down absorption spectroscopy is established for taking into account the laser linewidth effects. It is demonstrated that by fitting the logarithmic ring-down function to a truncated polynomial, absorption coefficent can be extracted from coefficients of the first as well as higher order terms of the fitted polynomial. The new fit model results in higher accuracy and broader dynamic range than the previous single exponential model. PMID- 15810293 TI - [Measurement for optical nonlinearity and optical bistability in II - VI semiconductor by optical waveguide method]. AB - The untransparent substrates on the epitaxial layer need not be etched out from chemistry etching to prepare the "window" through light. The optical absorption, optical nonlinearities and optical bistabilities of the semiconductor thin films with the nanostructure grown on the untransparent substrates can be directly measured by the end face coupling method of optical waveguide. Meanwhile, the several experiment examples of II - VI single crystal thin films and superlattices grown on GaAs substrate are demonstrated in this paper. PMID- 15810294 TI - [Anisotropic absorption spectra in YAlO3: (Nd3+ and Lu3+) single crystal]. AB - Absorption spectra of YAlO3: (Nd3+, Lu3+) single crystals for the three crystallographic axes were investigated at room temperature. The absorption band intensities distinctly revealed the anisotropy of YAlO3: (Nd3+, Lu3+). The Judd Ofelt theory was modified and applied to the treatment of the anisotropic absorption spectra. The absorption cross sections and the electric dipole oscillator strengths of various bands were calculated for three crystallographic axes of YAlO3: (Nd3+, Lu3+) crystals. Three sets of phenomenological parameters omegat(alpha) were determined. The validity of the modified Judd-Ofelt approach is discussed in detail. Moreover, we explained the anisotropy of the optical absorption of YAlO3: (Nd3+, Lu3+). PMID- 15810295 TI - [XPS study on the interaction of layers and pillars of layered double hydroxides pillared with heteropolyanion]. AB - Layered double hydroxides (LDHs) pillared with heteropolyanions were studied using XPS spectra. The shift of the binding energy of Mg2P, Al2P, Zn2P3/2, Cu2P3/2, Co2P3/2, N1s, Si2P, W4f7/2 and Mn2P3/2 as well as the mechanism of the interaction of layers and pillars were discussed. PMID- 15810296 TI - [The constant-in time DC component in the time-resolved FTIR spectroscopy]. AB - The direct current component in the interferogram of conventional FTIR spectroscopy is expressed by integral(infinity)0 B(sigma)dsigma. However, in time resolved FTIR spectroscopy, the direct current component Idc(tau) also depends upon tau, the time which IR signal appears. Idc(tau) is then represented by integral(infinity)0 B(sigma, tau)dsigma, called as constant-in-time DC component by us. Idc(tau) is proportional to the signal intensity B(sigma). Idc(tau) can not be filtered by any band pass filter. After Fourier transforming, the Idc(tau) converts to a strong peak at 0cm(-1) in the IR spectrum. We call it as constant in-time zero frequency signal Btau(0). Either Idc or Btau(0) can be used to monitor a variety of experimental conditions, such as the variation of gas pressure, laser intensity, the carbon deposition on the optics, etc. They can also be applied to measure the instrumental response time and the band pass frequency. PMID- 15810297 TI - [Studies on the oxidation reaction of octanol-2 with nitric acid by infrared spectroscopy]. AB - In this paper, the reaction process of oxidation of octanol-2 with nitric acid has been studied by IR spectroscopy. It is found that the main components of non sapoifiable matter are different in different oxidation degrees. The relation between oxidation products and the amount of nitric acid are investigated,the reaction mechanism has also been studied. Experimental results show that the oxidation process of octanol-2 is as follows: first, octanol-2 is oxidated to octanone-2, or to nitrate, nitrite and nitrile copmpounds, then these compounds are reoxidated to caproic acid in the meantime some by-products, such as valeric, enanthic acids are also found in oxidated products. PMID- 15810298 TI - [FTIR microscope study on the change of membrane protein and lipids of erythrocyte damaged by free radicals]. AB - The FTIR Microscope and Computer Aided Analysis were used to study not only the change of secondary structure of erythrocyte membrane protein attacked by the free radicals, but also the effect of oxygen centered free radicals on the membrane lipids in this paper. The results indicate that the content of alpha helix in the secondary structure of protein clearly changed because of the attack of the free radicals. Damaged by free radicals in 30 minutes, the protein structure could not be restored after 3 hours. The number of the P=O (vp=0), C=O and C=C in the lipids of the membrane was changed. These changes were caused by the peroxidation of the oxygen centered free radicals. PMID- 15810299 TI - [Study on the structures of poly (urea) urethane by FTIR and 1H NMR methods]. AB - Using the FTIR and 1H NMR methods, the structures, relative ratio and inventory ratio of segmented poly (urea) urethanes which are synthesized with different extenders (1,4-butandiol, ethylene diamine and their mixtures)have been studied successfully and the ratationship between the branch ratio and inventory ratio has been determined. PMID- 15810300 TI - [Spectroscopic characterization of a new terbium complex with red fluorescence]. AB - Usually, the emission spectra of terbium complexes exhibit 490 nm, 545 nm bands (green color) under the excitation of UV light, which is ascribed to the two 5D4 7F6, 5D4 - 7F5 transition. In this report, pink red fluorescence under the excitation of 300-370nm UV light is observed for a new 1, 10-phenanthroline-2 naphthoate complex of terbium. The fluorescence, FT-IR, ICP and X-ray fluorescence analysis were performed. The results showed that under the perturbation of the ligands field, the probability of 5D4 - 7F3 transition of terbium ion was greatly enhanced, while the probability of 5D4 - 7F5 transition was greatly decreased in the complex. Therefore, pink-red fluorescence was observed for the title complex. PMID- 15810301 TI - [Investigation on the luminescence properties of Er ions in Er-doped sol-gel silica glasses]. AB - Er-doped sol-gel silica glasses which have room temperature photoluminescence at 1.54 microm and doped concentration of 10(20)/cm(3) have been prepared. The examination results Indicated: (1) PL spectrum has largest intensity at the 0.5W% doped concentration; (2) the intensity of PL decreased 74%, when the temperature raised form 4K to 300K; (3) The Er ion and O ions formed complexes with coordinated numbers of 8 or 9. PMID- 15810302 TI - [Comparative study on the hydration of phosphate group in H2O/NaDEHP/n-heptane and H2O-TBP systems by FT-IR]. AB - The hydrations of phosphate group in both tributyl phosphate (TBP)-H2O system and water/NaDEHP/n-heptane reverse micellar system have been studied using Fourier transform infrared spectroscopy (FT-IR). Water content (W0 = [H2O]/[TBP]) in TBP solutions was measured quantitatively using FT-IR. In water/NaDEHP/n-heptane reverse micellar system, the stretching frequencies of phosphate group shifted from 1229 to 1200cm(-1) when water content increased from 1 to 25. For TBP-H2O system, P=O stretching band varied from 1281.33 to 1263cm(-1) with the increasing water content (from 0.33 to 1.55). P=O stretching bands in TBP-H2O system were also found to split into two peaks located around 1283 and 1260cm(-1) after the bands were resolved by curve-fitting. All these variations are attributed to the hydration of phosphate in the two systems. PMID- 15810303 TI - [FT-IR study on the solid-state reaction of bilirubin on BaF2]. AB - Bilirubinate complexes are the main component of pigment gallstones and the recent researches suggested that bilirubin free radical plays a role in some biological processes. In this paper the solid-state reaction of bilirubin on BaF2 was examined by FT-IR. The solid-state reaction was produced by grinding the crystalline bilirubin and single crystals BaF2 together in an agate mortar. From the results the free radical mechanism is suggested in the solid-state reaction of bilirubin on BaF2. The replacement of H on NH of pyrrole by Ba2+ was prior to that of H on COOH. The structure of the resultant complex from the solid-state reaction is different compared with that from the liquid-state reaction. PMID- 15810304 TI - [Analysis of the pollutant in purified diethyl malonate by on-line LG-GC/FTIR method]. AB - In some cases, the use of conventional GC/FTIR technique is limited due to its lower detectability to micro-components in organic mixtues in comparison to GC/MS. In this paper, an integration of LG/GC using retention gap technique with partially concurrent solvent evaporation, as a efficient chromatographic system for sample preconcentration and preseparation, was coupled into FTIR system in order to overcome the detectability problems of GC/FTIR analysis. The applicability of the on-line LG-GC/FTIR method is demonstrated by analysis of the isomers of divinylbenzene in purified diethyl malonate under conditions of hexane/dichloromethane (85:15) used as mobile phase in LC, solvent evaporation temperature of 75 degrees C and inlet pressure of 0.20MPa etc. PMID- 15810305 TI - [Correction of spectral interferences in inductively coupled plasma atomic emission spectrometry with a modified chemometric method based on matrix projection]. AB - A modified chemometric method based on matrix projection was developed to correct the spectral interferences in inductively coupled plasma atomic emission spectrometry (ICP-AES). The accuracy of the correction was significantly improved by reevaluating the residuals. The excellent analytical results for some rare earth elements proved the method fairly feasible. PMID- 15810306 TI - [Influences of Freon-12 (CF2CCl2) on the electron number densities in an inductively coupled plasma (ICP) measured with Halpha line--the applications of ICP in the disposal of hazardous waste]. AB - The inductively coupled plasma was applied to decompose Freon 12 (CF2CCl2). The efficiency of the decomposition was indirectly evaluated by comparing the electron number densities in the central channel of the ICP with the addition of different amounts of Freon-12. Halpha line (656.3nm) was used to measure the electron number density due to Hbeta line (486.1nm) is seriously interfered with the addition of Freon-12 under the selected operating conditions. The "thermal pinch" effect was found with the addition of Freon-12, which is beneficial to the complete decomposition of the compound. PMID- 15810307 TI - [Studies on the general properties of a novel microwave plasma enhanced glow discharge source]. AB - This paper introduced a novel microwave plasma enhanced glow descharge source, the structure design and operation were decribed, and the mutual effects of the main characters, such as pressure, current, voltage, microwave power and sputtering rates were also investigated in details. PMID- 15810308 TI - [Survey on recent progresses in glow discharge mass spectrometry]. AB - Glow discharge mass spectrometry as a sensitive method for direct solid sample analysis has gained rapid development in recent years. In this paper, the progresses of glow discharge mass spectrometry in the designment of new types of discharge lamp and the achievement in the diagnostic study of the atomization and ionization in the discharge process have been reviewed. Also the analytical characteristics of this method in practical application have been described. PMID- 15810309 TI - [GFAAS determination of trace manganese in human hair with graphite probe furnace]. AB - In this paper, direct determination of trace manganese is reported with graphite probe furnace atomic absorption spectrometry. It is shown that graphite probe furnace system has advantages in terms of accuracy, sensitivity, speed, simplicity. The proposed method has been applied to the determination of trace manganese in human hair with satisfactory results. PMID- 15810310 TI - [Spectral determination of the contents of elements in bean and bean-product]. AB - Determination of the contents of elements in Bean and Bean product using Z-8000 atomic absorption spectrometry is described. This results obtained indicate that high amounts of Mn, Fe and Mg are embodied in Bean and Bean-products. The Beans are good food for longevity and intelligence development. PMID- 15810311 TI - [Determination of bismuth in little sample by FAAS with home-made hydride generator]. AB - In order to determine Bi in little sample a simple hydride generator was designed, which was made easily and operated conveniently. The generator consumed sample of 0.5-1.0mL with the content of Bi of 10-100ng/mL, the precision, senstivity and detection limit were shown to be satisfactory. With this apparatus the Bi in little sample was determined by FAAS and the regression equation c = 30.17 + 190.6A, relation coefficient r = 0.9934, residual standard deviation of A for fixed c,SA.C = 2.737, standard error of regression SB = 4.001 and t-test result, where alpha = 0.001, showed that the equation was significant in statistics. The detection limit in this method was 3.69ng/mL. PMID- 15810312 TI - [Indirect determination of sulphur in plant tissues by flame atomic absorption spectrometry]. AB - The sulphur in plant tissues was transformed into SO4(2-) by digestion and it reacted on BaCrO4 to produce CrO4(2-) by exchange reaction, then the sulphur quantity in the plant was indirectly determined by flame atomic absorption spectrometry (FAAS). Various conditions affecting the quantative determination of SO4(2-) with this method were studied and the interference of PO4(3-), which existed in the digestion solution, could be overcome by addition of sufficient calcium. The results obtained by this method are consistent with those obtained by ICP-AES. PMID- 15810313 TI - [Determination of cadmium in tea by GFAAS]. AB - A method for the determination of cadmium with platform graphite furnace atomic absorption spectrometry was studied with PdC12 and Mg(NO3)2 as matrix modifier. The method is simple and accurate, and can eliminate the matrix effecte. It can be applied to directly determine cadmium in the tea sample. PMID- 15810314 TI - [The study and application of software (LLLZ) connecting the data processing programme with PW1404 X-ray fluorescence spectrometry (II)]. AB - The software LLLZ was applied to the analysis of geological powder sample. The original data could be used directly and quickly, and the analytical method could be chosen fast with this software, which was easily developed and used. PMID- 15810315 TI - [Synthesis and spectroscopic properties of neodymium and 3-methoxybenzoic acid with 1,10-phenantyroline]. AB - The complexes with the formula NdL3 x phen x H2O (phen = 1,10-phenathroline, HL = 3-methoxybenzoic acid) were synthesized and characterized by elemental analysis, molar conductivity, IR, UV-Vis and 1HNMR. PMID- 15810316 TI - [Spectrofluorometric determination of corticosterone in plasma and tissue]. AB - Corticosterone is one of the adrenal hormones. This paper reported the determination of corticosterone in plasma and tissue of small mammals with spectrofluorometry. The linear relationship between the fluorescence intensity and the concentration of corticosterone is over the range of 0.01-0.24 microg/mL (lambdaex = 470nm, lambdaem = 525nm). The average rate of standard recovery is 96.0% and RSD (n = 10) is 4.2%. This method is simple, rapid and sensitive. PMID- 15810317 TI - [The study of the colour reaction of new reagent N-nonyl-N'-(sodium p aminobenzenesulfonate) thiourea (NPT) on Pd(II) and its application]. AB - In this work, the colour reaction of Pd(II) on water-soluble spectrophotometric reagent N-Nonyl-N'-(Sodium p-Aminobenzenesulfonate) thiourea (NPT) which is newly synthesized has been studied. The result indicates that Pd(II) reacts on NPT form a yellow water-soluble complex with the proportion of 1:4 in HAc-NaAc buffer solution at pH5.8-6.6. The presence of CTMAB makes a synergistic sensitizing effect on the reaction,the complex exhibits a maximum absorption wavelength at 297.6nm, apparent molar absorptivity epsilon97.6 = 1.27 x 10(5) L x mol(-1) x cm( 1). Beer's law is obeyed when concentration of Pd is within the range of 2-11.2 microg/25 mL, 50 kinds of ions do not influence the determination. It shows NPT is one of the best reagents on its sensitivity and selectivity in the determination of Pd(II). This method obtains satisfying result when using in the determination of Pd in anode mud and catalyst. PMID- 15810318 TI - [Enrichment with polyamide and dual-wavelength spectrophotometric determination of gold and silver]. AB - The absorbent performance of polyamide resin has been studied. Au(III) and Ag(I) easily react with [SCN]- to form complex anions in dilute nitric acid medium, and be adsorbed on the polyamide resin to enrich, and be separated from many other base metals. When Na2SO3 and NH3 x H2O is used as eluating agent, Au and Ag can be recovered quantitatively. Therein can be retreated and reused easily. In addition, we established a dual-wavelength spectrophotometric method which has been applied to the determination of Au(III) and Ag(I) in standard samples with relative standard deviation of less than 5.1%. PMID- 15810319 TI - [Simultaneous determination of germanium and molybdenum in mixtures by complexation with 5'-nitrosalicyfluorone and dual-wavelength standard addition spectrophotometry]. AB - In the presence of hexadecyltrimethylammonium bromide and in a medium of 0.3 0.9mol/L sulfuric acid, germanium (IV) and molybdenum (VI) react with 2,6,7 trihydroxy-9 (2'-hydroxy-5'-nitro) phenylfluorone-3, i.e., 5' nitrosalicyfluorone, to develop red complexes, respectively. The spectra of the two complexes overlap seriously each other, and the additivity of their absorbances is good in the wavelength range of 490-548nm. So a new method for simultaneous spectrophotometric determination of germanium and molybdenum by dual wavelegth standard addition method has been investigated in detail. The analytical results of five standard mixtures are more accuracy than those obtained by isobestic-point dual-wavelength method. The method has been applied to the analysis of traditional chinese medicine and mineral water with satisfactory results. PMID- 15810320 TI - [Spectrofluorimetric study of trifluoroacetylacetone (TFA)-gadolinium(III)-CTMAB system and application]. AB - The determination of microamount gadolinium(III) by fluorescence method was studied based on complex formation Gd(III)-CTMAB-TFA at pH7.2. Beer's law is obeyed in the range of 0. 005-0.04 microg/mL Gd(III). The detection limit is 0.003 microg/mL. The method is used to determine microamount gadolinium of rare earth mine with satisfactory result, and compared with that of ICP-AES method. PMID- 15810321 TI - [Spectral studies of vitamin K3 in aqueous solution by pulse radiolysis and laser photolysis]. AB - The absorption spectra of VK3 in aqueous solution were studied by pulse radiolysis and laser photolysis technique, and the formation mechanism of intermediates of transient species was suggested, which gave the examples for studying the mechanism of blood-clotting of VK3 and provided the theoretic evidence for further understanding the reasons of VK3 as a radiosensitizer. PMID- 15810322 TI - [Study of the circular dichroism for tribasic complex of amine derivative of beta cyclodextrindien-Cu2+ -benzoic acid]. AB - The circular dichroism spectra of beta-cyclodextrindien, beta-cyclodextrindien Cu2+ complex, beta-cyclodextrindien-benzoic acid complex, tribasic complex of beta-cyclodextrindien-Cu2+ -benzoic acid were determined by using JASCO J-20C automatic recording spectropolarimeter. The attribution of the bands obtained were made according to the order of orbit energy of Cu2+ complex and benzoic acid, and the coordination structure of tribasic complex of amine derivative of beta-cyclodextrindien-Cu2+ -benzoic acid were decided acording to the structure character of Cu2+ complex and KAJART sector rule of inclusion complex of beta cyclodextrindien with aromatic compounds. PMID- 15810323 TI - [Spectroscopic characteristics of Ce3+ doped Ca3Al2M3O12 (M=Si, Ge) garnets]. AB - The spectral properties of Ce3+ doped Ca3Al2M3O12 (M = Si, Ge) garnets have been reported under excitation of UV light. The emission spectra of Ce3+ ion in garnets consist of a broad band assingning to the 5d-4f transition of Ce3+ ion. The emission intensity Ce3+ ion in Ga3Al2Ce3O12 shows very weak, but in Ga3Al2SiO12 shows strong. The emission intensity of Ce3+ ion is greatly enhanced by adding Gd3+ or flux into Ca3Al2Si3O12: Ce garnet. At 295 and 77K, the fluorescence decay of Ce3+ ions in Ca3Al2Si3O12 garnet have been measured and the fluorescence lifetimes of Ce3+ ions are 46 and 49ns, respectively. PMID- 15810324 TI - [Spectral study of podophyllotoxin and its derivatives]. AB - In this paper, we have studied all kinds of static absorption spectra and transient absorption spectra of podophyllotoxin and its derivatives. We have located the site of absorption spectra and their forming mechanism with spectrofluorometry, Ultraviolet, pulse radiolysis and laser photolysis, and suggested the connection of their structrue with their anti-tumour and toxicity. PMID- 15810325 TI - [Analysis of synchronous and derivative synchronous fluorescence spectra of phenol and O-dihydroxybenzene]. AB - In this paper, the analyses of the synchronous flourescence spectra of phenol and O-dihydroxybenzene by quasi-square linear dual combination of multiwavelength data dual-peak multiplication balance, multiwavelength linear regression, first and second order derivative technique are proposed. Five new methods for simultaneous determination of these compounds are constructed. The accuracies and recoveries of the proposed analytical methods are found to be satisfactory. PMID- 15810326 TI - [Cetyltrimethylammonium bromide fluorescence enhancement of aspirin]. AB - The fluorescence intensities of aspirin with cetyltrimethylammonium bromide enhanced by micellar solutions have been examined. It is found that fluorescence enhancement depend on the concentration of CTAB and pH of solution. PMID- 15810327 TI - [Spectral study of the conformational change of yeast alcohol dehydrogenase induced by alcohol]. AB - The conformational change of Yeast Alcohol Dehydrogenase (YADH) at the different concentration of ethanol, n-propanol and ethylene glycol was studied by means of ultraviolet spectrum, fluorescence spectrum and circular dichroism spectrum. The results showed that the ultraviolet absorbance at 220nm and 280nm as well as the relative fluorescence intensity at 336nm of YADH increased with increasing alcohol concentration. The negative peakes at 208nm and 220nm of YADH in circular dichroism spectrum with the solvent of ethanol, ethylene glycol were obviously intensified, but the 220nm peak of YADH was increased in the presence of n propanol while the 208nm peak was decreased and red-shifted in position as to completely lost. According to the data above, it indicates that the conformation of YADH was changed with losing activity at the various concentration of alcohol. PMID- 15810328 TI - [The interior relation between FDP and SHG]. AB - The fluorescence-disappearing phenomenon (FDP) was used due to the nonlinear interaction between the laser and the matter, a universal phenomenon. It was often neglected during the research of surface-harmonic generation (SHG). Many experiments showed that there was a interior relation between FDP and SHG. In this paper, the mechanism of surface-harmonic generation has been discussed in detail, according to the fluorescence-disappear viewpoint and the fluorescence disappear model (FDM) proposed also. The new mechanism was summarized by following three points: (a) The multiphoton absorption generated visible fluorescence spot, (b) Laser-heating effect and other effects promoted the conversion from spontaneous emission into quasi-stimulated emission, (c) Being accompanied by the fluorescence disappear, the part of laser energy which was initially used to cause fluorescence was now converted into developing the harmonic signal. PMID- 15810329 TI - [Room temperature phosphorimetric determination of practolol with dansyl chloride labeling]. AB - Dansyl Chloride (DNS-Cl) was used as a phosphorescent labeling agent for the determination of practolol. The derivative was formed by refluxing practolol ethyl acetate solution with the solid DNS-Cl in the presence of potassium carbonate anhydride. Room-temperature phosphorescent (RTP) properties of the derivative were studied by a micelle-stabilized room-temperature phosphorescence (MS-RTP) method. Phosphorescence lifetimes of practolol-DNS and the DNS-Cl were found to be 0.791 and 0.408ms, respectively. By applying the derivative MS-RTP technique, practolol could be quantitatively determined without previous separation process. Detection limit of practolol with the proposed method was 0.19 microg/mL. PMID- 15810330 TI - [Analysis of air pollutants with a combination of GC/FTIR and GC/MS]. AB - In this paper, the air pollutants were analyzed with a combination of Gas chromatography/Fourier transform infrared spectroscopy (GC/FTIR) and Gas chromatography/Mass spectroscopy (GC/MS). The organic mixture was well separated after a careful adjustment of GC and IR parameters in GC/FTIR. In the identification of unknown components, MS is a main method, while IR is also useful to confirm the results. PMID- 15810331 TI - [The characteristic infrared spectra of alpha,alpha'-dioxoketene cyclic N,N acetals]. AB - The characteristic infrared spectra of alpha,alpha'-dioxoketene cyclic N,N acetals have been studied and the changing rules with the change of chemical structure have been discussed. PMID- 15810332 TI - [The ultraviolet spectra of alpha,alpha'-dioxoketene cyclic S,S-acetals(N,N acetals)]. AB - In this paper, the ultraviolet spectra of alpha,alpha'-dioxoketene cyclic S,S acetals (N,N-acetals) have been studied and their changing characters with the change of chemical structure have been indicated. PMID- 15810333 TI - [Infrared spectroscopy application in soil organic matter]. AB - As an important method to study the constitution and properties of macromolecular organic compounds, the infrared spectroscopy has been more and more widely taken in the researches of soil organic matters (SOM). Especially,the application of FTIR and the combined uses of FTIR with chromatogram etc. have made the researches of SOM get a great progress in many aspects. In this paper, the infrared spectroscopy applications were reviewed in SOM. It includes the following contents: the methods to study SOM by IR, studies on the constitution of soil humic substances (SHS), extraction of SOM and classification of SHS, decomposition, transformation and humification of organic matters, the differences of SOM in different situations, the interactions of SHS with metais, clay minerals and other organics in soil. PMID- 15810334 TI - [Some physical-chemical properties of the colloid of bentonite in Xinjiang Tuoksun county]. AB - The X-ray diffraction pattern, infrared spectrum, differential thermal curve, dewatering curve, cation exchange capacity and specific surface of the colloid of bentonite in China (Xinjiang Tuoksun) are presented and are compared with that of bentonites in the united states (Wyoming), New Zealand (Quarget) and French. The results indicate that particles in China are finer, crystals are better, purities are higher and capabilities of absorbing water are greater. So that it has better technology characteristic in metals. PMID- 15810335 TI - [Determination of trace rare earth impurities in the high-purity lanthanum oxide by chemical spectrographic analysis]. AB - The determination of trace rare earth impurities in the high-purity Lanthanum oxide by chemical spectrographic analysis is reported in this paped. The matrix, La2O3, was separated by the extraction-eluating resin P507. The remained rare earth impurities were absorpted by carbon powder. The rare earth impurities absorpted were determinated in carrier KBH4 by spectrographic analysis. The trace rare earth impurities in 99.9999% La2O3 can be determinated by this method. PMID- 15810336 TI - [Determination of Sn/In in the ITO film by emission spectral method]. AB - In the film sample analysis, the consstence within standard sample and analytical sample is discussed by emission spectral method. Using spark light source and moving electrode, the content of Sn/In in the ITO film is obtained by repeatedly exposing four samples. The accuracy and precision are satisfying. PMID- 15810337 TI - [Determination of non-rare earth impurities including Cu, Fe, Ca and Ni in samarium oxide by ICP-AES]. AB - The direct determination of non-rare earth trace impurities including Cu, Fe, Ca and Ni in high purity Samarium oxide by means of ICP-AES is presented in this paper. By using orthogonality experimentation, we studied main factors of influential determination and selected compromise condition of simultaneous determination of Cu, Fe, Ca and Ni. Influence of matrix concentration was discussed. We used the method of matrix match and subtraction background. For the sample solution containing 5mg/mL of Sm2O3 the determination limits were 0. 0003% for Cu, 0.003% for Ni, 0.001% for Ca and Fe, respectively. The relative standard deviation was in the range of 2.6-9.2%. The recovery ratio of these elements was in the range of 90-110%. The method is simple and rapid. It was fit to productive control analysis and determination of production. PMID- 15810338 TI - [Determination of beryllium in geological samples with slurry sampling and probe atomization in graphite furnace atomic absorption spectrometry]. AB - A method of loading a slurry prepared from a solid sample for analysis of beryllium in geological samples using graphite furnace atomic absorption spectrometry was approached by combining slurry sampling with probe atomization. The factors that influence the performances of beryllium including ashing and atomization temperature, atomization surfaces (probe and tube), the materials of the tube and probe, and the use of matrix modifiers were investigated. The results show that the determination of beryllium in geological samples could be performed by direct slurry sampling with the use of calcium as the matrix modifier for a calibration established with aqueous standards. The relative standard deviations (RSD) of the method were 3.1-5.2%. The analytical results of the geological reference samples were consistent with the certified values. The activation energy during beryllium atomization was calculated using the Arrhenius graph method which was in good with the bonding energy of Be-O. PMID- 15810339 TI - [Respective determination of inorganic germanium and germanium-132 in foods]. AB - Inorganic germanium and carboxyethyl germanium sesquioxide (germanium-132) in health drinks were respectively determined by hydride generation-atomic fluorescence spectrometry (HG-AFS). The conditions of respective determination of inorganic germanium and germanium-132 in natural foods were preliminarily discussed. PMID- 15810340 TI - [Determination of trace Al in serum of cancer patients by graphite furnace atomic absorption spectrometry using K2Cr2O7 and surfactant as matrix modifier with molybdenate coated tubes]. AB - Different kinds of graphite tubes and of matrix modifiers have been tested for optimization of operation condition for determination of Al in serum by GFAAS. The effects of different interference ions have been eliminated by adding K2Cr2O7 and Triton X-100 as matrix modifier. The method is suitable for determination of serum of cancer patients. The characteristic mass is 21.15pg/0.0044 A x S. The recovery rate is 104.92. The relative standard deviation is less than 8.17%. The result obtained is satisfactory. PMID- 15810341 TI - [Determination of trace copper and lead in highly pure samaria by extraction graphite furnace atomic absorption spectrometry]. AB - In this paper, we have studied the meassuring condition and the extracting condition for the determination of trace copper and lead in highly pure samaria by extraction-GFAAS. Trace copper and lead in highly pure samaria were determined. The relative standard deviations are less than 6%, the recovery is 98 101% for copper and 93-100% for lead. The method is simple, accurate and practical. PMID- 15810342 TI - [A rapid Zeeman graphite furnace atomic absorption spectrometic method for the determination of trace lead and cadium in drinking water]. AB - The determination of trace lead and cadium in drinking water by rapid GFAAS has been described in this paper, which is based on eliminating ashing step, shortening drying time and without chemical matrix modifier. The fast speed and simplicity makes it possible for routine analysis. PMID- 15810343 TI - [Determination of zinc, copper, iron and manganese contents in hair for MPA patients and healthy men]. AB - In this paper the contents of zinc, copper, iron and manganese in hair for MPA patients and 38 hair-healthy men have been determined by FAAS. The results show that the hair zinc and manganese levels in MPA patients are significantly lower than that in hair-healthy men. The hair copper level in MPA patients is significantly higher than that in hair-healthy men. There are statistic difference (P < 0.05) in hair zinc, copper and menganese levels among two groups. There is no statistic difference (P > 0.05) in the iron level among the two gopups. This paper provides useful material for the research of the cause and treatment of the MPA. PMID- 15810344 TI - [Determination of aluminium in healthy person's hair by FAAS and FAES]. AB - In this paper, two methods of N2O/C2H2 FAAS and FAES are applied to determine aluminium in healthman hair. The requirement of determination, interferences of inorganic ions and organic substances and the results of determination are contrasted respectively. It is shown experimently that the interference ratio of substances, accuracy, sensitivity and recovery ratio of FAES are superior to FAAS at 396.1 nm wavelength in acidity medium. The RSD (n = 9) of FAES and FAAS are 2.2 and 2.7% respectively and detect limit is 0.07 microg/mL for FAES and 0.25 microg/mL for FAAS. The recovery ratio of adding 20.0 microg aluminium to 1.0 g hair samples are 96.5-105.0% and 93.5-106.5% respectively. PMID- 15810345 TI - [Chemical shifts of CrKbeta peaks in XRF spectra by Si(Li) spectrometry]. AB - The energies of CrKbeta X-Rays in metel Cr, Cr2Os and K2CrO4, which are excited respectively by gamma-ray 59.6keV from radioactive source 241Am, have been measured by Si(Li) spectrometry. It is shown in the experiment that the peak of CrKbeta X-ray in K2CrO4 shifts 2.12 +/- 0.27eV compared to that in metel Cr. PMID- 15810346 TI - [Chemical state analysis of chromium by the conventional X-ray fluorescence spectrometer]. AB - X-Ray Kbeta emission spectra of chromium and some of their compounds were studied by conventional XRF spectrometer Rigaku 3080 E1 and PEAKFIT spectrum analyzing programs. Some qualitative and quantitative relations were obtained between the true spectrum profile and its parameters (peak position, peak top intensity, FWHM and asymmetry indices) and chemical state by data processing technique. And making use of these relationship, the chemical state distribution were analyzed of chemical chromate treatment film, stainless steel. Satisfactory results were obtained. PMID- 15810347 TI - [An application of genetic algorithms to the analysis of EXAFS spectrum]. AB - Extended X-ray absorption fine structure (EXAFS) has been extensively used in the structural study in recent years. But the current methods of analysis EXAFS spectrum is difficult and tedious,demanding either very similar standard samples or the initial parameters near the solution. In this paper, a new technique Genetic Algorithms (GAs), was applied to the analyzing of the EXAFS spectrum,and two spectra of CU samples were investigated by the method. It was found that reasonable results can be obtained by the method only with limiting the initial parameters to a broad range. Comparing the results with those obtained by EXCURVE88, the fitting is superior, and the parameters are reasonable. Therefore, it is proved that the GAs may be a high performance method for the analyzing of the EXAFS spectra. PMID- 15810348 TI - [Determination of Ni and Pd in white karat gold jewellery by the EDXRF extrapolate-regression method]. AB - The EDXRF extrapolate-regression method described in this paper combines regression method with the fundamental formula of fluorescence intensity. The contents of Ni and Pd in white karat gold jewellery were calculated theoretically according to the spectrum of the sample. The content of gold was deternined without standards. The precision was 0.1% and the deviation was 0.3% compared with AA. PMID- 15810349 TI - [Measuring trace germanium in vegetables with the spectrofluorometric method]. AB - A spectrofluorometric method to measure trace germanium in vegetables was established. Results showed that the detection limit of the method was 0.569 ng/g, the accuracy of determination on the 1 microg Ge content was +/- 3.59%. The content of Ge in vegetables could be calculated from the standard adding method without extraction when using mixed acid digestion. PMID- 15810350 TI - [The choice of optimum instrumental parameters of a new type interference selective amplitude modulation spectrometer (SISAM) with monograting]. AB - A specific calculation of a family of spread functions of the new type SISAM with only one grating, which is equal in effect to two grating, is made as a function of the dimension of light source and the modulation amplitude and their optimum values are determined. PMID- 15810351 TI - [New application of atomic absorption spectrophotometer in testing technology]. AB - A simple and satisfactory method is developed for the new application of AAS. We can just rely on atomic absorption spectrophotometer without flame on place of UV Visible spectrophotometer. The operation is very simple, and it can raise the efficiency of AAS. This experiment offers a possibility to find new applications of atomic absorption spectrophotometer. PMID- 15810353 TI - [Studies of magnetic circular dichroism and absorption spectra in alpha-NisO4 x 6H2O crystal]. AB - Alpha-NiSO4 x 6H2O crystals were studied by magnetic circular dichroism (MCD) and absorption spectra measurements in the range lambda = 185-3300nm. The results show that there are three aborption bands, of which the peaks are related to the d-d transitions from the ground state to the excited states. The weak peaks of a progression of the three members were observed between the principal structures lambda = 654nm and lambda = 713nm due to spin-orbit coupling interacion. The weak peaks of a progression of four members were also observed around the new absorption peak at lambda = 585nm in MCD spectra. PMID- 15810352 TI - [Spectral study on the self-polymerizing reaction of N-diisopropyl phosphoryl amino acids]. AB - By using polarization sensitive CARS method, five kinds of N-diisopropyl phosphoryl-alpha-amino acids are investigated. They polymerize themselves to form peptide at 55 degrees C for 40 hours. The vibration mode spectra prior and after the reaction are recorded. With the help of theoretical calculation, the configuration of these peptides are inferred to be trans form and the specificities vibration modes are assigned. PMID- 15810354 TI - [Photoluminescence and UV-Vis absorption spectra characteristics of metal complexes of 8-hydroxyquinoline]. AB - 8-hydroxyquinoline complexes of Na(I), Ca(II), Mg(II), Zn(II), Cu(II), Mn(II), Al(III), In(III) and La(III) were synthesized. Photoluminescence (PL) and UV-Vis absorption spectra characteristics of these complexes and 8-hydroxyquinoline are reported. Except complexes of Cu and Mn, most of these complexes reveal photoluminescence. With the polarization force of metal cations increasing, the PL peak shows red-shift and two broad absorption peak in UV-Vis (300-400nm) absorption spectra distinctly appears apart. PMID- 15810355 TI - [Determination of trace rare earth in fossil of dinosaurian egg by laser thermal lens spectrometry]. AB - Determination of trace rare earth in fossil of dinosaurian egg by LTLS (Laser thermal lens spectrometry) is presented in this paper. The trace rare earth in fossil of dinosaurian egg was separated and concentrated with ion exchange method. The measure conditions and effect factors are discussedian. The LTLS was applied to the determination of trace rare earth in fossil of dinosaurian egg with satisfactory results. It provides the reference data for the studies on the fossil of dinosaurian egg and an analytical method for the determination of rare earth in geochemical sample. PMID- 15810356 TI - [X-ray photoelectron spectroscopy (XPS) and mass spectrum (MS) study of organic tin compounds II]. AB - Seven new synthetical (Z)-1-[2-(Triarylstannyl) vinyl]-1-cycloheptanols organic tin compounds were studied by X-ray photoelectron spectroscopy and mass spectrum method. The influence of different replacement radical in compounds on the d orbit of tin atom was discussed. The coordinate substance had an effect on the chemical displacement of Ols binding energy. The O-->Sn bond was affected by the different replacement radical. The results of X-ray photoelectron spectroscopy and mass spectrum show that Cl, Br and I had the important action. PMID- 15810357 TI - [Study on the arylaminothiourea compounds by IR spectroscopy]. AB - In this paper, eleven new thiourea compounds were prepared and studied by IR spectroscopy. The fundamental vibrations have been assigned. The band near 1540cm(-1) was identified as thiourea I band, which mainly originated from the N H deformation vibration coupling with a little C-N stretching vibration. Thiourea I band near 1280cm(-1) mainly originated from C-N stretching vibration. C=S stretching vibration frequency was at near 110cm(-1) and it was identified as thiourea III band. PMID- 15810358 TI - [Study on syntheses and IR spectra of novel chelate resins containing hydroxyalkylamine group]. AB - Two novel chelate resins containing hydroxy-alkylamine group were prepared. The IR spectra of the above mentioned resins and their middle products were measured, and the major IR bands were assigned by such chemical methods as hydrolysis, getling salts, determination of hydroxyl value, etc., the structure of the resins and their middle products were proved. PMID- 15810359 TI - [Study on the interaction of metal ion in active center of superoxide dismutase with cobalt chloride by visible spectroscopy]. AB - The direct interaction of metal ion in active center of copper-zinc superoxide dismutase (Cu2Zn2SOD) with inorganic cobalt chloride (CoCl2) in buffer has been first studied by visible spectroscopy. The effects of increasing amounts of CoCl2, changing pH,different acid salt and equilibrium time on the interaction have been discussed. It was found that the Co(II) of CoCl2 makes an exchanging interaction with Zn(II) in Cu2Zn2SOD and thus the partial Cu2Co2SOD is formed. PMID- 15810360 TI - [Resonance Raman spectra of transition metal Mn(III), Fe(III), Rh(III), Pd(II) porphyrin complexes]. AB - The resonance Raman spectra of Mn(III), Rh(III), Fe(III), Pd(II) protoporphyrin IX-dimethylester (PP) and tetra-phenylporphyrin (TPP) complexes were reported. The difference of their structure-sensitive band frequency was a result of the interaction of different metal ions' outer d electrons with the electrons of the porphyrin ring. As well the effects of different porphyrin rings and their substituts on the structure-sensitive bands were discussed in this paper. PMID- 15810361 TI - [The study of interactions of [CpFe(Co)2]2 with surface of alumina and Titania supports by Fourier transform diffuse reflection and photoacoustic infrared spectroscopy]. AB - The diffuse reflection and photoacoustic infrared spectra of the species derived from dicarbonyl (eta5-cyclopentadienyl)iron (0) {[CpFe(CO)2]2 (Cp = eta5-C5H5)} which are supported on the surfaces of acidic,neutral and basic alumina (Al2O3) and Titania (TiO2) have been measured. The structures, stabilities and concentrations of these supported organometallic species depend markedly on the acid-base properties of the alumina surfaces. There is mainly species of [CpFe(CO)2Fe-H-Fe(CO)2Cp]+ on the surface of [CpFe (CO)2]2/Al2O3 (acidic) (A). The species of CpFe(CO)2(-O-) and [CpFe(CO)]4 are observed on the surface of [CpFe(CO)2]2/Al2O3(neutral)(B). On the suface of [CpFe(CO)2]2/Al2O3 (basic) (C), the mainly species is [CpFe(CO)2]4 with a few CpFe(CO)2(-O-) species. The concentration of species is highest on the surface of A and lowest on the, surface of C. The diffuse reflection and photoacoustic (middle and near )infrared spectra of [CpFe(CO)2]2/TiO2(T) show that the [CpFe(CO)2]2 absorbed on the surface of TiO2 is stable in the air at the room temperature. There is no a species derived from [CpFe(CO)2]2 to be observed. The diffuse reflection near infrared spectra of [CpFe(CO)2]2(D) and [CpFe(CO)2]2/TiO2 (T) are measured. Most of them are assigned to overtone and combination of carbonyl vibrations and several peaks are assigned to triple overtone and triple combination. The diffuse reflection infrared spectroscopy is the good method to study the interaction of organometallic compounds with metal oxide powder. PMID- 15810362 TI - [Surface-enhanced Raman spectra studies on four complexes derived from D glucosamine and alpha-glycine and their effect on DNA]. AB - Raman scattering of four complexes derived from d-glucosamine and alpha-glycine and their effects on DNA were studied with surface-enhanced Raman spectra (SERS). It was found that their SERS had shown little difference for their similar structural. Their interaction abilities on DNA were poor according to the change of their SERS spectrun after adding DNA to their water solution except for Co(III)GluG, which has strong effect on DNA. NiGluG's adsorptive ability on Ag sol is the strongest among other three complexes. PMID- 15810363 TI - [Study on the properties and microstructure of the gel in the extracted organic phase containing DMHPA-RE(III)]. AB - The content of Na+, RE3+ and the swelling characteristic of dried gel containing DMHPA[di-(1-methylheptyl)phosphoric acid]-RE(III) have been studied by FTIR and ICP/AES spectroscopy methods. The result demonstrates that (1) the mechanism of microemulsion extracting lanthanide ions is complicated and it is difficult to be explained with simple chemical equations, (2) The dried gel can swell, but the aggregation states and microstructure change as the used solvents and its content vary, (3) The far infrared spectroscopy shows that there are ring structures in the gel and differences of the ability in coordination with DMHPA between light and heavy lanthanide ions. PMID- 15810364 TI - [Study on FTIR spectra of HDEHP and DMHPA in different states]. AB - FTIR spectra of pure di (2-ethylhexyl) phosphoric acid (HDEHP), di (1 methylheptyl) phosphoric acid (DMHPA) and their sodium salts, microemulsions, extracted organic phases containing rare earth were studied. It was seen that the microstructures of the two extractants changed with their different alkyl structures and aggregation states. PMID- 15810365 TI - [Enhanced emission spectra of Yb, Eu, Y and Sm HCLS with high current microsecond pulsed supply]. AB - The emission spectra of Yb, Eu, Y, Sm hollow cathode lamps have been investigated with a high current microsecond pulsed (HCMP) supply developed in our laboratory. Both atomic and ionic emission lines of interest of these rare earth elements are enhanced by tens to tens of thousand times in HCMP mode compared to DC mode. Effects of parameters of the supply on emission intensities are surveyed. HCMP HCLs may be used as an economic and convenient excitation source for atomic/ionic fluorescence spectrometric analysis. PMID- 15810366 TI - [Artificial neural network applied for spectral overlap interference correction in ICP-AES]. AB - A back-propagation artificial neural network (BP-ANN) has been applied to correcting spectral overlap interference in inductively coupled plasma atomic emission spectrometry (ICP-AES). Some network parameters including the range of input values and training sequence for training patterns presented to the network were discussed using simulated Ce 413.380nm and Pr 413.380nm line profiles. Results show that the noise in simulated mixture spectra will slow down the network convergence and has more influence on network prediction. PMID- 15810367 TI - [Quantitative determination of the nickel, tin, copper, scandium, zirconium and hafnium in silicate sample by laser microspectrochemical analysis]. AB - Quantitative determination of the nickel, tin, copper, scandium, zirconium and hafnium in silicate sample were carried out by laser microspectrochemical analysis, results show that the method has good precision and accuracy. The stability of the laser output and the selection of auxiliary spark excitation condition in experimental process were discussed. PMID- 15810368 TI - [The determination of high content tin in powder samples by line width method]. AB - The quantitative determination of 60% Sn in tantalum-tin mineral samples was carried out by line width method. When the concentrations of internal standard element Pb in all three standard series was 1%, 2%, and 3% respectively,the influence of Pb to the analytical results was discussed. The experimental results show that the different contents of the internal standard are different for both the accuracy and precision of spectral analysis. PMID- 15810369 TI - [Simultaneous determination of 16 elements in decoction solution of Chinese herbal medicine by ICP-AES]. AB - A new method has been developed for simultaneous derermination of 16 elements in decoction solution of Chinese herbal medicine by ICP-AES after pretreatrment the solution with HNO3-H2O2. This method has low and stable blank value. The detection limits of 0.1-5ng/ml for most elements and recoveries of 85-103% were obtained. This method has been applied to the determination of 16 elements in decoction solution of chinese herbal medicine with satisfactory reswls. PMID- 15810370 TI - [Rapid spectrochemical qualitative analysis of simple OMA system]. AB - Simple optical multichannel analyzer (OMA) system which is formed by ordinary one dimension CCD camera, spectrograph and microcomputer was introduced in this paper. Spectrochemical analysis feasible of this OMA system was studied. The applied software programed, by us can realize wavelength calibration, spectral line identification and give out automatically the results of qualitative and semidefinite quantity analysis. PMID- 15810371 TI - [Determination of Pb in flying coal-ash by GTAAS]. AB - Biochemical was chosen as the stability and improvement reagent to make insoluable solid into slurry, which could be sampled directly to determine Ph in flying coal-ash, one of the environmental samples. The optimal conditions of Hitichi GTAAS 180-80 were decided by experiments, such as linear range 7.0 66.7ng/mL atomization temperature 2000 degrees C and amount of biochemical 0.2 1.2 per thousand. The effects of acid, stirring and particle concentration were also addressed. PMID- 15810372 TI - [Hydride generation-AFS determination of trace arsenic, antimony, selenium and mercury in Chinese herbal medicine in Qinghaixizang plateau]. AB - In this paper, a hydride generation non-dispersive atomic fluorescence spectrometry method has been developed for the determination of trace arsenic, antimony, selenium and mercury in Chinese herbal medicine in Qinghai-Xizang plateau. Under the optimum condition, the quantities of fourteen interference elements are below certain non-interferent limits. The detection limits were as follow (g/mL): As 1.3, Sb 0.39, Se 0.24 and Hg 0.44. The relative standard deviation of 9 determination was As 2.4-4.2%, Sb 2.5-5.6%, Se 3.1-5.0% and Hg 3.8 5.9%. The recovery of four elements was 94.7-105.4%. This method is simple, rapid, sensitive and accurate, and has been successfully tested with As, Sb, Se and Hg in more than 20 kinds of Chinese herbal medicine in Qinghai-Xizang plateau. PMID- 15810373 TI - [Determination of chromium, manganese and nickel in six biological agents]. AB - The determination of chromium, manganese and nickel in biological agents has obtained by flame atomic absorption spectrometry. The results show that three trace elements of body necessary are the most rich in ant factor injection liquid. The chromium content is slightly lower in the brain cell activator injection liquid, the nickel content is slightly lower in the shift factor injection liquid and the manganese is the lowest in the marrow factor injection liquid. In this paper, we have studied three trace element contents in six biological agents. It provides, useful data for clinical treatment to some trace element-lacking diseases. PMID- 15810374 TI - [The study and application of software (LLLZ) connecting the data processing programme with PW1404 X-ray fluorescence spectrometry(I)]. AB - The structure, flow chart, hardware disposition and main function of the software LLLZ, which connecting the data processing programme with the PW1404 X-ray fluorescence spectrometer, was briefly introduced. PMID- 15810375 TI - [Simultaneous spectrophotometric determination of three-component mixture by two partial least squares methods]. AB - Two partial least squares methods, classical partial least squares (PLS) and partial least squares based on a kernel algorithm (KPLS), were studied for simultaneous determination of a three-component mixture. Three programs called SPGRAFA, SPGRPLS and SPGRKPLS were designed to perform the calculations. Eight error functions were calculated for deducing the number of factors. Because the size of the kernal matrix was much smaller than the original data matrix, the KPLS applied to calculating the matrix with many wavelengths and fewer number of samples. Experimental results showed both methods to be successful even there was overlap of spectra and agreed well. PMID- 15810376 TI - [Determination of trace iron and cobalt in natoral water by ammonium thiocyanate resin phase-spectrophotometry]. AB - At the presence of Tween-80 in acid medium, Fe(III) Co(III) and SCN- are changed into coloured complexations and quantitatively absorbed on the resin. The apparent molar absorptivities of resin phase are epsilon Fe = 2.4 x 10(5)L x mol( 1) x cm(-1) at 490nm and epsilon Co = 2.1 x 10(5)L x mol(-1) x cm(-1) at 630nm by spectrophotometry. Beer's law is obeyed by Fe(III) and Co (III) in the range of 0 8microg/25mL and 0-15microg/25mL. method has been applied to natural water. Relative standard deviations are 1.7% and 5.5%, respectively. PMID- 15810377 TI - [Study on cleavage mechanisms of dimer (t-Bu)2NO in several solvent phases]. AB - In this paper, we studied the cleavage products of dimer t-BuNO in aqueous and organic solutions using method of 1HNMR and UV-Vis, analyzed the reactive kinetics, put forward that dimer (t-Bu)2NO homolytically cleavaged in nonpolar organic solution, and that dimer (t-Bu)2NO homolytically and heterolyticall cleavaged in a competitive manner in polar aqueous solution. PMID- 15810378 TI - [Strain effects in Zn1-xMn(x)Se superlattices of II -VI diluted magnetic semiconductors]. AB - We report the strain effects of Zn1-xMn(x)Se superlattices of II -VI diluted magnetic semiconductors by the photoluminescence measurements with compostions x = 0.2, 0.3, 0.4 in the temperature range T = 11K to 300K. The results show that the exciton energy tends to a red shift with the increasing of x due to the strain effects. The distribution of strain in wells and barriers presents a considerable change when their thickness ratio is altered. Photoluminescence spectra peaks as a function of temperature are mainly dominated by the band gap of ZnSe. The temperature coefficient of exciton energy is obtained. PMID- 15810379 TI - [Study on the interaction between CP and DNA]. AB - In this paper, interaction between CP (Cis-ddp or ciplatin) and DNA (deoxyribonucleicacid) was studied using the fluorescence probe as well as the absorption spectra of DAN. PMID- 15810380 TI - [Study on the TICT property of phenothiazine and its derivatives]. AB - The radiationless quantum yields ( phi(nr) ) of phenothiazine (PTH). N-phenyl phenothiazine (PHZ). N-(2-pyridine) phenothiazine (PYZ) and model compound p-N, N dimethylaminobenzonitrile (DMABN)in benzene were measured. The effects of solvents on phi(nr) of DMABN,PHZ,PYZ and PTH were investigated respectively. It was found that phi(nr) of PYZ and DMABN were dependent on the solvent polar parameter (E(T)(30)) . From the results of radiationless quantum yields and combining the state and dynamic fluorescence spectra, it is proved that PYZ as well as DMABN appears TICT property. PMID- 15810381 TI - [The synthesize of mono-( 6-S-DNS )- beta-CD and its molecular recognition ability]. AB - Mono- ( (6-S-DNS) - beta -CD is synthesized and characterized. The interaction between the luminescent derivative of beta-cyclodextrin and guest (cyclohexanol, cyclohexene, Bromocyclohexane, and p-phthalic acid, m-phthalic acid 3, 5 Dinitrobenzoic acid, p-hydroxybenzoic acid, p-animobenzoic acid)has been slutied. The binding constants and sensitivety factors are obtained based on modified Hildbrand-Benesi equation. The results show that the guest-Host inclusion complex with 1:1 stoichiometry is formed and that the binding constants and sensitivity factors for molecular recognition is directly related to the molecular structure and hydrophobic character of guest. PMID- 15810383 TI - [Fluorescence studies on the mixed solid complexes of europium-terbium-1, 10 phenanthroline-phthalate systems]. AB - The 1, 10-phenanthroline-phthalate complexes of europium and terbium have been synthesized. Two series of mixed complexes, mechanical ground complexes (series 1) and chemical synthesized with mixed EuCl3 and TbCl3 solutions (series 2), have been also prepared. Fluorescence studies on all of the samples were performed. The results of their fluorescence spectra excited by 365nm UV-light showed that there are interactions between europium ions and terbium ions in these two kinds of mixed complexes. Their fluorescence intensities strongly changed especially for chemical synthesized mixed complexes, in which the fluorescence intensities of europium ions were greatly sensitized by terbium ions and those of terbium ions were strongly quenched by europium ions. But the peak positions of their emission bands did not shift obviously. The structure of these complexes have been characterized by FT-IR spectra. The results suggested that no free COOH groups exist and all of the nitrogen atoms of 1, 10-phenanthroline molecules coordinate to rare earth ions in all of these complexes. PMID- 15810382 TI - [The medium effects on fluorescence of coumarin-1 complexes]. AB - The fluorescence spectra of 4-methyl-7-diethylaminocoumarin (coumarin-1) in cyclohexane-alcohol, 1, 4-dioxane-water binary mixed solvents and cyclodextrin, surfactant aqueous solutions were studied in detail. The reasons that fluorescence spectrum of coumarin-1 is very sensitive to environmental polar change were discussed based on solvent-solute molecule hydrogen bond and its effects on formation of nonradiative twisted intramolecular charge transfer (TICT) state for the fluorescence probe. In the meanwhile, it' s inclusion constant with CD and bonding constant with surfactan micella were determined. PMID- 15810384 TI - [Study of the photoluminescence spectra of laser dye impregnated in porous silicon]. AB - We studied the photoluminescence (PL) of the compound films, which were made by implanting some laser dyes into the porous silicon films, and compared it with the PL of the laser dyes in other states. We also measured the PL of the compound films with different substrates. It was found that the spectra symmetry of the implanted laser dyes is better than that of the dyes in other states. The experimental results suggest that the PL of the compound films mainly comes from the monomers of the implanted laser dyes. PMID- 15810385 TI - [Surface photovoltaic spectroscopic investigation of Bacteriorhodopsin (BR)]. AB - The surface photovoltaic spectra (SPS) of Bacteriorhodopsin-based films were systematically measured. The results show that the BR films are characterized by the specific photovoltaic effect. The SPS response of the BR films depends on the fabrication and humidity of the film. The intermediate M412 and O640 of BR are observed in the SPS of HEPES-containing BR film, indicating that HEPES has very important function to stabilize intermediate M412. The observations are explained in terms of the photocycle of BR and the mechanism of photovoltage generation. PMID- 15810386 TI - [Fluorometric determination of trace Shachongdan with calcein-Pd2+]. AB - A fluorometric determination of trace Shachongdan (SCD) [(CH3)2NHCH (CH2S2O3- )CH2S2O3Na x H2O] is described. The method is based on the calcein display fluorescence in pH6-7 phosphate buffer solution, it react with Pd2+ to the fluorescence disappeared. The complex of SCD-Pd2+ is more stability than the complex of calcein-Pd2+, when SCD react with calcein -PdZ+, the calcein can be freed and its fluorescence appears once again. The fluorescence excitation and emission wavelenghts are 494 and 514nm. The linear range is 2 x 10(-7) mol/L-3 x 10(-6)mol/L and the detection limit is 6 x 10(-8)mol/L. PMID- 15810387 TI - [Study on fluorescence method of pefloxacin]. AB - Pefloxacin is a new kind of antibiotic. In this paper, a sensitive fluorescence method has been developed for the analysis of Pefloxacin. It has stronger fluorescence excitation. The fluorescence of Pefloxacin was measured at excitation wavelength 282nm and emission wavelength 440nm. The method is highly sensitive with lower limit of detection, calibration curves were linear between 1 x 10(-9) - 1 x 10(-6)mol/L, with a regression coefficient of 0.9999. At the same time, recoveries of pefloxacin in Human serum Albumin and Urine are 95% and 105% respectively. PMID- 15810388 TI - [Molfig, a software for displaying the structure and vibration of molecule]. AB - A Molfig software has been developed for displaying the structure and vibrational mode of molecule in our lab. Various functions and a friendly interface are equipped in the software. The testing results showed that the software may enhance our understanding of the relationship between the vibrational behavior and the structure of molecules. PMID- 15810389 TI - [Spectroscopic study of Langmuir-Blodgett films of octa-4-(hexadecoxy) lanthanium bisphthalocyanine]. AB - The surface pressure/molecular area isotherms ( pi approximately A ) of lanthanium bisphthalocyanine were investigated, showing an edge-on molecular orientation of the macrocycle rings on the water subphase. Langmuir-Blodgett films of this compound were prepared and characterized using IR, UV-vis absorption spectroscopy. PMID- 15810390 TI - [IR and UV-vis spectra of nitrogen-doped fullerene]. AB - Nitrogen-doped fullerene derivative's IR and UV-vis spectra are reported. Some of the C60 IR silent modes are activated due to nitrogen atom reducing C60 molecular symmetry. UV-vis spectrum indicates nitrogen is donor. PMID- 15810391 TI - [Study on the orientation of PET three-dimensional crimp hollow fiber by IR-ATR]. AB - The variations of structure and orientation of poly (ethylene terephthalate) (PET) three-dimensional crimp hollow fiber were investigated by infrared attenuated total reflection (IR-ATR) method. It was found that the carbonyl group of this PET hollow fiber orientates vertically to the fiber surface. PMID- 15810392 TI - [The infrared spectra of alkyl hydrogen styrylphosphonates and their complexes with metals]. AB - The infrared spectra in the range of 4000 approximately 650cm(-1) for alkyl hydrogen styrylphosphonates (HA (SP)), 2-ethylhexyl hydrogen 2 ethylhexylphosphonate and di-2-ethylhexyl hydrogen phosphate and their complexes with metals were recorded. The tested HA (SP) included n-octyl, 2-ethylhexyl, 1 methylheptyl and sec-alkyl (C11-13) hydrogen styrylphos- phonates. The metals studied were Fe ( III ), Cr ( III ), In ( III ), Nd ( III ), Cu ( II ), Co ( II ), Ni ( II ), Ca( II ) and Mg( II ). In the spectra of these organophosphorus compounds adsorption at approximately 1450cm(-1) is present only in HA (SP). The Spectra of HA (SP) show the broad absorption of hydrogen bonded OH at approximately 2200 and 2600cm(-1) ( vOH ). For each of the organophosphorus monoacids upon complexation with the metals excepting Ca and Mg, the vOH disappears and the v(P=O) shifts to a lower frequency , wthereas in the cases of Ca and Mg the OH, P=O and P--O-- absorption bands all become broader. PMID- 15810393 TI - [Structural characterization of copper-phthalocyanine thin solid films by FTIR spectroscopy]. AB - The structure of tris- (2, 4-di-t-amylpheoxy) - (8-quinolinoxy) copper phthalocyanine (CuPc) thin solid films has been characterized by Fourier transform infrared (FTIR) transmission, polarized transmission and reflection absorption (RA) spectroscopy. The following conclusions can be obtained from the above measurements: (1)in LB films, the hydrocarbon chains of CuPc are in hexagonal or pseudohexagonal subcell packing, the CH2 asymmetric vibrational vector is oriented with respect to the substrate surface and the RA spectroscopy can distinguish the two CH2 streching modes of benzene cycle; (2)in sublimed films, the molecules of CuPc are out of order. PMID- 15810394 TI - [Fluorescence and FT-IR spectroscopic studies on the mixed solid complexes of europium and terbium phthalate]. AB - In this investigation, the fluorescence and FT-IR spectroscopic studies on synthesized Eu-Tb phthalate complexes and mixed solid complexes of europium and terbium phthalates were perfomed. The experimental results of the emission spectra excited by 365nm UV-light showed that the emission peak positions of Eu ( III ) [or Tb ( III )] wer shifted slightly due to the interaction between Tb ( III )and Eu ( III ) ions. However, the fluorescence intensities of one rare eath ion Eu ( III ) [or Tb (III )] were strongly influenced by the other. FT-IR spectra indicated that the coordination structures of carboxyl groups, both in mechanically mixed and synthesized solid complexes, were also changed. These results suggested that there are interactions between Eu ( III ) and Tb ( III ) ions in their mixed complexes. PMID- 15810395 TI - [FT-IR study of Ce3+-doped calcium phthalate phosphor]. AB - FT-IR of the CaC8H4O4 : Ce phosphor prepared by the solution reaction method were determined. Bands of their spectra were assigned. Some structure information about these compounds were given. PMID- 15810396 TI - [Study on atomic emission spectrum and spectral interferences of rare earth elements with a high resolution inductively coupled plasma atomic emission spectrometer.--II. Spectral intererences of Samarium matrix]. AB - The rare earth elements play a very important role in today's manufacture and life. For their complex spectra it is difficult to determine the trace elements in rare earth element matrix. This work investigated the spectral interferences of samarium matrix on other rare earth elements with a high resolution inductively coupled plasma atomic emission spectrometer. A lots of spectrum information and spectral interferences were obtained, which are quite useful in the selection of analytical lines that free of spectral interferences and the elimination of the concomitant's interferences. PMID- 15810397 TI - [Progress in determination of rare earths by atomic/ionic fluorescence spectrometry]. AB - The progress of determination of rare earths by atomic/ionic fluorescence spectrometry is reviewed. The experimental systems and the analytical characteristics of the technique are briefly discussed, and its applications are illustrated in detail. The research trends are predicted as well. The systems with appropriate combinations of excitation sources and atomization/ionization cells are well proved to increase analytical sensitivity and minimize interferences. For the excitation soures, hollow cathode lamp has been proved to be economic and practical, although pulsed dye laser has been intensively investigated. The inductively oupled plasma is a preferable atomization/ionization cell. The atomic/ionic fluorescene spectrometry is one of effective techniques for the determination of rare earths mainly due to its low spectral interference and wide linear dynamic range. PMID- 15810398 TI - [Recent advances in absolute analysis by graphite furnace atomic absorption spectrometry]. AB - This paper summarizes recent advances in absolute analysis by graphite furnace atomic absorption spectrometry, including the developments of absolute analysis in atomic absorption spectrometry, the theoretical calculation of characteristic mass, the study of the stability of experimental characteristic mass and the applications of standardless analysis in various samples. PMID- 15810399 TI - [Determination of the trace silver by FAAS after separation and preconcentration with cotton cellulose xanthate]. AB - Silver was absorbed by cotton cellulose xanthate in solution at pH 6, and was eluated quantitatirely from CCX with 3% HNO3 solution and determination by FAAS. The method is simple and rapid and has been apllied to the determination of silver in rocks and water. PMID- 15810400 TI - [X-ray fluorescence spectrometrlc determination of nine heavy rare earth elements in the enriched mixture of thulium oxide, ytterbium oxide and lutecium oxide by asming thin film technique]. AB - A method for the determination of nine heavy rare earth elements in the enriched mixture of thulium oxide ytterbium oxide and lutecium oxide by XRF method has been studied in this poper. The samples were prepared by ashing thin film. This method is characterized as it is low cost, economic and simple in standards preparation, and standards can be easily preserved. The accuracy and precision of the results were satisfactory for the demands of prodnction. PMID- 15810401 TI - [Determination of caffeine and sodium benzoate contents in caffeine and sodium benzoate injection by P-matrix method]. AB - The contents of caffeine and sodium benzoate injection may be determined by P matrix method without separation of its components. BSAIC language is used for programming. The average recovery of caffeine is 100.0% and that of sodium benzoate is 99.95%. The variation coefficient of caffeine is 0.40% and that of sodium benzoate is 0.65%. On the basis of recovery rate and data observed it is concluded that this method is accurate, reliable, simple and rapid as compared with the standard pharmacopoeia regulations. PMID- 15810402 TI - [The alignment of the optical system for 216 coude focus echelle spectrometer]. AB - This paper gives a brief introduction about the structure of the echelle spectrometer which was installed at coude focus of the chinese 2. 16 meter astronomical telescope. According to the design requirment of this echelle spectrometer, the main points and steps of alignment of optical system are analysed. Authors work out a practical alignment scheme in which the fewest auxiliary tools are used. PMID- 15810403 TI - [A spectroscopic method using mixed states]. AB - We introduce a method by use of atomic mixed states for the study of atomic spectroscopy. PMID- 15810404 TI - [Study on the quantum yields of triplet state formation]. AB - The quantum yields of triplet state formation ( phiT ) of acridine, benzophenone. phenothiazine and phenanthrene were measured with anthracene as calibrating compound by the means of pulsed laser photoacoustic calorimetry. It was found that phiT of these compounds were not in accordance with the literature results except acridine due to slow response of PZT transducer. But the better results could be obtained by using deconvolution method to eliminate the effect of respose of the transducer. PMID- 15810405 TI - [Energy pooling collisions between excited Na and K atoms]. AB - Two different atomic species, sodium and potassium, are resonantly excited in a ceel by a CW dye laser and a potassium lamp, respectively. The density of the excited Na (3P) atoms in the cell is measured directly using the absorption equivalent-width technique on the lines of the diffuse series. To isolate the heteronuclear contribution in the fluorescence spectrum, a modulation detection technique is adopted. The energy-pooling collisions between Na (3P) and K (4P) are observed and cross sections are obtained. PMID- 15810406 TI - [Investigation on absorption edge and Ep, Eg of tetragonal crystal system SBN]. AB - In this paper, the station of the absorption edge of Sr0.63 Ba0.37 Nb2O6 single Crystal is measured with experimental studies. The absorption coefficient is calculated with transimitting curve. The absorption edge below 3.5eV is attributable to indirect transition by studing the alpha1/2-hv curve. The phonon energy Ep and the energy gap Eg are ascertained. PMID- 15810407 TI - [Spectra of FLMR of 14N 16OX 2pi3/2R(1. 5) v =0-->1 and 15N 16OX 2pi3/2Q(1. 5) v = 0-->1 transition]. AB - The apparatus of Faraday effect LMR used for the detection of 14N 16O and 15N 16O is described in this paper. We gave and analyzed the spectra of FLMR of 14N 16OX2pi3/2 (1. 5) v =0-->1 transition and 15N 16OX2pi3/2Q(1.5) v =0-->1 transition. The dependence of FLMR signal on the sample molecular concentration has been stadied. PMID- 15810408 TI - [The potential energy function of the internal rotation for H2S2 molecule]. AB - Using the perturbation theory without wave function, the analytical formula of the internal Rotation energies has been derived, and the potential energy function of internal rotation for H2S2 molecule has been determined using the published data for the torsional frequencies. PMID- 15810409 TI - [Potential energy function of internal rotation for the molecules of (CX3)2Y type]. AB - Studing on the molecules having M -tops of N -fold symmetries, decoupling the equation of motion for rotation and using the perturbation theory without wave function, the formula of the energies of internal rotation has been derived and the potential energy functions of internal rotation for (CH3)2O and (CF3)2CO have been determined. PMID- 15810410 TI - [Ending diatomic molecular terms]. AB - A method is discussed to find diatomic molecular terms of equivalent and non equivalent configuration. PMID- 15810411 TI - [The sawtooth oscillation phenomenon of visible spectral signal in HT-6M Tokamak]. AB - The sawtooth oscillation phenomenon of visible spectral signal in HT-6M Tokamak is presented. The influences of electron temperature, electron density and atomic ground density on the spectral signal discussed. This phenomenon results mainly from the change of electron temperature at the edge. PMID- 15810412 TI - [The characteristic infrared spectra of alpha,alpha'- dioxoketene cyclic dithioacetals]. AB - In this paper, based on the results of previous paper[1], the other characteristic infrared spectra of alpha,alpha'- dioxoketene cyclic dithioacetals have been studied and their changing rules with the change of chemical structure have been indicated. PMID- 15810413 TI - [Study on self-activation of phosphorylated amino acids to form peptides by FTIR]. AB - The self-activation of six phosphorylated amino acids to form peptides was studied in i-propyl alcohol by FTIR. The results indicated that the phosphorylated amino acids could be self-activated to form peptides in alcohol and the FTIR method in the study on the self-activation of phosphorylated amino acids to form peptides can provide useful information for life process. PMID- 15810414 TI - [TR-FTIRS study of crystal phase transformation in polyvinylidene fluoride]. AB - The study of polyvinylidene fluoride (PVDF) by time-resolved Fourier transform infrared spectroscopy (TR-FTIRS) was reported. The instantaneous change and process of both alpha-form and beta-form during transformation were studied under the pulse electric field excitation. Transformation of alpha-form to beta-form was indicated by the increasing of the peak at 508cm(-1), the decreasing of the peak at 528cm(-1) and a shift of those peaks. The transient change time was about 5ms. The decreasing of the peak at 528cm(-1) was nonlinear. A hysteresis-like loop was observed, which was related with the biaxially orientated property of PVDF and was in good agreement with its polarizated hysteresis loop. PMID- 15810415 TI - [Nondectructive analysis of liquid samples with near-infrared spectroscopy]. AB - This paper studied the near-infrared spectra of liquid samples. The plots of weights at each wavelength showed the spectral characteristics of pure materials. The samples could be classified by using plots of the scores of their first principal components. The principal component regression of samples was made. The obtained results were identical with the reference analytical values. The advantages of near-infrared spectroscopy, combined with principal component analysis, lie in its nondectructivity for samples and suitability for on-line measurements. PMID- 15810416 TI - [Curvefit, an overlapping bands resolving software for IR spectra]. AB - A Curvefit software has been developed for resolving the overlapping band of the vibartional spectra in our lab . Various functions and a friendly interface are equipped in the software. The testing results showed that both the speed of the calculation and the reliability of the final results are satisfactory. PMID- 15810417 TI - [Far FT-IR spectra of gallium hexafluoride ions]. AB - Far FT-IR spectra of Rb3GaF6, alpha-Cs3GaF6 and high temperature form Cs3GaF6 and their XDS have been studied in this paper. The results indicated that Rb3GaF6 and high temperature form Cs3GaF6 are tetragonal, a = 0.632,c = 0. 886nm and a = 0.687,c = 0. 987nm respectively. alpha-Cs3GaF6 is orthorhombic, a = 1. 062,b = 0. 996,c = 0. 561nm PMID- 15810418 TI - [A resonance Raman spectrometer with UV-visible continuously tunable excitation lines]. AB - A set of resonance Raman spectrometer with excitation lines continuously tunable in 220-970nm, which is a first set in China, has been assembled. The factors influenced on the signal noise ratio (SNR) of spectra were analyzed in detail. The methods to improve the SNR were suggested and the parameters of the spectrometer were optimized. A satisfied Raman spectra can be obtained. PMID- 15810419 TI - [Application of UV spectroscopy in structural studies of metal centre of metal protein]. AB - The principle of UV spectroscopy applied in structural studies of metal centre of complexes is introduced in this paper. Several examples prove that such spectroscopy is an effective technical method in structural studies of metal centre of metal-protein. PMID- 15810420 TI - [Sinultaneous determination of rare earth in rare earth ore band tea-leaves by ICP-AES]. AB - A method for simultaneous determination of rare earth elements in tea-leaves by ICP-AES is described. The results and their standard deviations obtained from the analysis of tea-leaves are summarized. The recovery rate of 16 rare earth elements is 88.7-106.9%. The RSD is <8%. PMID- 15810421 TI - [Determination of calcium in brains of rat by ICP-AES]. AB - This paper reports the determination of calcium in brains of rat by ICP-AES. The recovery rate is 97%, the RSD is 1.16%, and the results are satisfactory. The interference effects of phosphor on the calcium under various operating conditions in ICP-AES were studied. PMID- 15810422 TI - [Determination of six trace elements in steels by ICP-AES with the changed internal standard]. AB - This paper studies the direct determination of P, Si, Mn, Cu, Cr and Ni in steels with the changed internal standard. The standard curves are standardized with FULL STANDARDIZATION, and the results of the sample are calibrated with TYPE STANDARD in time. The signal intensity influenced by RF power, observation height and sample gas folw is discussed. Results obtained for the determination of every element in steels are satisfactory. The RSD is less than 0.5% and the recovery is about 96-103%. The detection limit is about 0. 002-0.07%. PMID- 15810423 TI - [Constant temperature atomization in electrothermal atomic absorption spectrometry]. AB - The recent progrees of experimental techniquee and research of fundamental procees including kinetic and thermodynamic methods for atomization under isothermal conditions in GFAAS are reviewed in this paper, meanwhile, the limitations in currently available approaches are also diecussed. It covers 45 papera. PMID- 15810424 TI - [Determination of cadmium, cobalt, nickel in grain with Graphite Furnace Atomic Absorption Spectrometry]. AB - In this paper, studied the suitable operating conditions for determination of Cd, Co, Ni in grain by Graphite Furnace Atomic Absorption Spectrometry. This method has beer used to determine the three elements in grain and the results are satisfactory. PMID- 15810425 TI - [The change of trace elements content in hair during the period of gestation]. AB - Study on trace elements content change in hair during the period of gestation for 52 lying-in women was performed. The quantitative results for 7 elements have been obtained and the change tendencies for other 7 elements without standard value have been observed on the basis of a self comparison method for each individual. PMID- 15810426 TI - [Development of a SIRI-TOFMS and the application research progress]. AB - The only one set of Sputter Initiated Resonance Ionization-Time of Flight Mass Spectrometer (SIRI-TOFMS) in China was designed and developed. A liquid gallium ion source with micron level diameter of beam, frame-style piezoeletric ceramic sub-micron sample manipulator, a secondary electron imaging system and a computer controlled precise targeting system as well as a real colour big screen image displaying system were added so as to improve its microanalysis capability. By means of this apparatus, trace analysis of Au in minerals and Cu in standard steel samples were carried out. The detection limit of them is 40ng/ and microg/g level, respectively. Aside from the above detections, other preliminary researches ere also performed. PMID- 15810427 TI - [Study of interaction between cytochrome C and cystine using synchronours fluorescence spectroscopy]. AB - The interaction of cytochrome C with cystine as promoter was studied by synchronous fluorescence spectroscopy. After cystine was added into cytochrome C solution, the synchronous flurorescence spectra of tyrosine and tryptophan residues in cytochrome C were monitered with time. The results indicated that the lysine residues in cytochrome C molecules combine with cystine slowly. It induced the small conformational change of cytochrome C molecules. PMID- 15810428 TI - [Study on time-resolved fluorimetry of samarium]. AB - In this paper, a simple and highly sensitive laser fluorimetric system is described, and the value of time discrimination for suppressing unwanted signals and decreasing the minimum detectable concentration is demonstrated. It is confirmed that time discrimination is very useful for ultratrale determination of fluorescent complex. This technique is successful with the combined use of a pulsed laser and a pulse-gate signal detection system for samarium complex. The linear range and the detectin limit of the method are 0. 01-10microg/ml and 3.3ng/ml, respectively. The RSD for Sm3+ (0. 5microg/ml) ( n =12) is 3.7%. The method was used to detemine samarium in rare earth mixtures. The recovery was in the range of 90.0-110.0%, and the mean recovery was 101.4% ( n = 6 ) PMID- 15810429 TI - [Influences of terraazaindene on absorption spec tra and fluorescence spectra of sensitizing dyes]. AB - This study is about the influences of tetraazaindene (TAI) on absorption spectra and the fluorescence spectra of three kinds of sensitizing dyes. The absorption of all peaks of the dyes enhance with increasing TAI, among which the absorption of J-aggregate has even marked enhancement. At certain value of TAI, the absorption of monomolecule of dye enhances much more rapidly as well as the absorption of H-aggegte decreasing because of H-aggregate dissociating into monomolecules. The results obtained also show that the fluorescence emissions of the dyes enhance with increasing TAI. The mechnisms of the observed phenomena are discussed in detail. PMID- 15810430 TI - [Determination of Keganmin tablet with multi-wavelength K-factor spectrophotometry]. AB - The contents of phenacetin, aminopyrine and caffeine in Keganmin tablet were determined simultaneonsly with multiwavelength K -factor spectrophotometry. The result is accurate and the method is rapid. The average recoveries and relative standard deviations of the analysis for phenacetin, aminopyrine and caffeine were 99.91%, 0.41%; 100.1%, 0.29%; 100.1%, 0.56%, respectively. PMID- 15810431 TI - Automated metabolic reconstruction for Methanococcus jannaschii. AB - We present the computational prediction and synthesis of the metabolic pathways in Methanococcus jannaschii from its genomic sequence using the PathoLogic software. Metabolic reconstruction is based on a reference knowledge base of metabolic pathways and is performed with minimal manual intervention. We predict the existence of 609 metabolic reactions that are assembled in 113 metabolic pathways and an additional 17 super-pathways consisting of one or more component pathways. These assignments represent significantly improved enzyme and pathway predictions compared with previous metabolic reconstructions, and some key metabolic reactions, previously missing, have been identified. Our results, in the form of enzymatic assignments and metabolic pathway predictions, form a database (MJCyc) that is accessible over the World Wide Web for further dissemination among members of the scientific community. PMID- 15810432 TI - Genomic comparison of archaeal conjugative plasmids from Sulfolobus. AB - All of the known self-transmissable plasmids of the Archaea have been found in the genus Sulfolobus. To gain more insight into archaeal conjugative processes, four newly isolated self-transmissable plasmids, pKEF9, pHVE14, pARN3 and pARN4, were sequenced and subjected to a comparative sequence analysis with two earlier sequenced plasmids, pNOB8 and pING1. The analyses revealed three conserved and functionally distinct sections in the genomes. Section A is considered to encode the main components of the conjugative apparatus, where two genes show low but significant sequence similarity to sections of genes encoding bacterial conjugative proteins. A putative origin of replication is located in section B, which is highly conserved in sequence and contains several perfect and imperfect direct and inverted repeats. Further downstream, in section C, an operon encoding six to nine smaller proteins is implicated in the initiation and regulation of replication. Each plasmid carries an integrase gene of the type that does not partition on integration, and there is strong evidence for their integration into host chromosomes, where they may facilitate intercellular exchange of chromosomal genes. Two plasmids contain hexameric short regularly spaced repeats (SRSR), which have been implicated in plasmid maintenance, and each plasmid carries multiple recombination motifs, concentrated in the variable regions, which likely provide sites for genomic rearrangements. PMID- 15810433 TI - In vitro selection of an archaeal RNase P RNA mimics natural variation. AB - Archaeal and bacterial RNase P RNAs are similar in sequence and secondary structure, but in the absence of protein, the archaeal RNAs are much less active and require extreme ionic conditions for activity. To assess how readily the activity of the archaeal RNA alone could be improved by small changes in sequence, in vitro selection was used to generate variants of a Methanobacterium formicicum RNase P RNA: Bacillus subtilus pre-tRNA(Asp) self-cleaving conjugate RNA. Functional variants were generated with a spectrum of mutations that were predominately consistent with natural variation in this RNA. Variants generated from the selection had cleavage rates comparable to that of wild type; variants with improved cleavage rates or lower ionic requirements were not obtained. This suggests that the RNase P RNAs of Bacteria and Archaea are globally optimized and the basis for the large biochemical differences between these two types of RNase P RNA is distributed in the molecule. PMID- 15810434 TI - Interactions between RNase P protein subunits in archaea. AB - A yeast two-hybrid system was used to identify protein-protein interactions between the ribonuclease P (RNase P) protein subunits Mth11p, Mth687p, Mth688p and Mth1618p from the archaeon Methanothermobacter thermoautotrophicus. Clear interactions between Mth688p and Mth687p, and between Mth1618p and Mth11p, were confirmed by HIS3 and LacZ reporter expression. Weaker interactions of Mth687p and Mth688p with Mth 11p, and Mth11p with itself, are also suggested. These interactions resemble, and confirm, those previously seen among the homologs of these proteins in the more complex yeast RNase P holoenzyme. PMID- 15810435 TI - Transaldolase of Methanocaldococcus jannaschii. AB - The Methanocaldococcus jannaschii genome contains putative genes for all four nonoxidative pentose phosphate pathway enzymes. Open reading frame (ORF) MJ0960 is a member of the mipB/talC family of 'transaldolase-like' genes, so named because of their similarity to the well-characterized transaldolase B gene family. However, recently, it has been reported that both the mipB and the talC genes from Escherichia coli encode novel enzymes with fructose-6-phosphate aldolase activity, not transaldolase activity (Schurmann and Sprenger 2001). The same study reports that other members of the mipB/talC family appear to encode transaldolases. To confirm the function of MJ0960 and to clarify the presence of a nonoxidative pentose phosphate pathway in M. jannaschii, we have cloned ORF MJ0960 from M. jannaschii genomic DNA and purified the recombinant protein. MJ0960 encodes a transaldolase and displays no fructose-6-phosphate aldolase activity. It etained full activity for 4 h at 80 degrees C, and for 3 weeks at 25 degrees C. Methanocaldococcus jannaschii transaldolase has a maximal velocity (Vmax) of 1.0 +/- 0.2 micromol min(-1) mg(-1) at 25 degrees C, whereas Vmax = 12.0 +/- 0.5 micromol min(-1) mg(-1) at 50 degrees C. Apparent Michaelis constants at 50 degrees C were Km = 0.65 +/- 0.09 mM for fructose-6-phosphate and Km = 27.8 +/- 4.3 microM for erythrose-4-phosphate. When ribose-5-phosphate replaced erythrose-4-phosphate as an aldose acceptor, Vmax decreased twofold, whereas the Km was 150-fold higher. The molecular mass of the active enzyme is 271 +/- 27 kDa as estimated by gel filtration, whereas the predicted monomer size is 23.96 kDa, suggesting that the native form of the protein is probably a decamer. A readily available source of thermophilic pentose phosphate pathway enzymes including transaldolase may have direct application in enzymatic biohydrogen production. PMID- 15810436 TI - Description of Thermococcus kodakaraensis sp. nov., a well studied hyperthermophilic archaeon previously reported as Pyrococcus sp. KOD1. AB - A hyperthermophilic archaeal strain, KOD1, isolated from a solfatara on Kodakara Island, Japan, has previously been reported as Pyrococcus sp. KOD1. However, a detailed phylogenetic tree, made possible by the recent accumulation of 16S rRNA sequences of various species in the order Thermococcales, indicated that strain KOD1 is a member of the genus Thermococcus. We performed DNA-DNA hybridization tests against species that displayed high similarity in terms of 16S ribosomal DNA sequences, including Thermococcus peptonophilus and Thermococcus stetteri. Hybridization results and differences in growth characteristics and substrate utilization differentiated strain KOD1 from T. peptonophilus and T. stetteri at the species level. Our results indicate that strain KOD1 represents a new species of Thermococcus, which we designate as Thermococcus kodakaraensis KOD1 sp. nov. PMID- 15810437 TI - The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly. AB - The signal recognition particle (SRP) RNA helix 6 of archaea and eukaryotes is essential for the binding of protein SRP19 and the assembly of a functional complex. The conserved adenosine at the third position of the tetraloop of helix 6 (A149) is crucial for the binding of protein SRP19 in the mammalian SRP. Here we investigated the significance of the equivalent adenosine residue at position 159 (A159) of Archaeoglobus fulgidus SRP RNA. The A159 of A. fulgidus and A149 of human SRP RNA were changed to C, G or U, and fragments containing helix 6 or helices 6 and 8 were synthesized by run-off transcription with T7 RNA polymerase. The ability of recombinant A. fulgidus and human SRP19 to form ribonucleoprotein complexes was measured in vitro. The simultaneous presence of A149 and helix 8 is required for the high-affinity binding of SRP19 to the human SRP RNA. In contrast, A. fulgidus SRP19 binds to the SRP RNA fragments with high affinity irrespective of the nature of the nucleotide, demonstrating that A159 does not directly participate in protein binding. Instead, as indicated by the resistance of the wild-type A. fulgidus RNA towards digestion by RNase A, this residue allows the formation of a tightly folded RNA molecule. The high affinity between A.fulgidus SRP 19 and RNA molecules that contain both helices 6 and 8 suggests that A159 is likely to initiate archaeal SRP assembly by forming a conserved tertiary RNA-RNA interaction. PMID- 15810438 TI - A proposal to rename the hyperthermophile Pyrococcus woesei as Pyrococcus furiosus subsp. woesei. AB - Pyrococcus species are hyperthermophilic members of the order Thermococcales, with optimal growth temperatures approaching 100 degrees C. All species grow heterotrophically and produce H2 or, in the presence of elemental sulfur (S(o)), H2S. Pyrococcus woesei and P. furiosus were isolated from marine sediments at the same Vulcano Island beach site and share many morphological and physiological characteristics. We report here that the rDNA operons of these strains have identical sequences, including their intergenic spacer regions and part of the 23S rRNA. Both species grow rapidly and produce H2 in the presence of 0.1% maltose and 10-100 microM sodium tungstate in S(o)-free medium. However, P. woesei shows more extensive autolysis than P. furiosus in the stationary phase. Pyrococcus furiosus and P. woesei share three closely related families of insertion sequences (ISs). A Southern blot performed with IS probes showed extensive colinearity between the genomes of P. woesei and P. furiosus. Cloning and sequencing of ISs that were in different contexts in P. woesei and P. furiosus revealed that the napA gene in P. woesei is disrupted by a type III IS element, whereas in P. furiosus, this gene is intact. A type I IS element, closely linked to the napA gene, was observed in the same context in both P. furiosus and P. woesei genomes. Our results suggest that the IS elements are implicated in genomic rearrangements and reshuffling in these closely related strains. We propose to rename P. woesei a subspecies of P. furiosus based on their identical rDNA operon sequences, many common IS elements that are shared genomic markers, and the observation that all P. woesei nucleotide sequences deposited in GenBank to date are > 99% identical to P. furiosus sequences. PMID- 15810439 TI - Hydrolase and glycosynthase activity of endo-1,3-beta-glucanase from the thermophile Pyrococcus furiosus. AB - Pyrococcus furiosus laminarinase (LamA, PF0076) is an endo-glycosidase that hydrolyzes beta-1,3-glucooligosaccharides, but not beta-1,4-gluco oligosaccharides. We studied the specificity of LamA towards small saccharides by using 4-methylumbelliferyl beta-glucosides with different linkages. Besides endo activity, wild-type LamA has some exo-activity, and catalyzes the hydrolysis of mixed-linked oligosaccharides (Glcbeta4Glcbeta3Glcbeta-MU (Glc = glucosyl, MU = 4 methylumbelliferyl)) with both beta-1,4 and beta-1,3 specificities. The LamA mutant E170A had severely reduced hydrolytic activity, which is consistent with Glu170 being the catalytic nucleophile. The E170A mutant was active as a glycosynthase, catalyzing the condensation of alpha-laminaribiosyl fluoride to different acceptors. The best condensation yields were found at pH 6.5 and 50 degrees C, but did not exceed 30%. Depending on the acceptor, the synthase generated either a beta-1,3 or a beta-1,4 linkage. PMID- 15810440 TI - [Multiresidue analysis of organochlorine pesticides and the application to Chinese herbal medicine]. AB - [A review of study on multiresidue analyses of organochlorine pesticides(OCPs) was presented here, which included sample extraction, clean-up and determination. The multiresidue analytical methods applied to Chinese herbal medicine was also discussed. PMID- 15810441 TI - [Advances on biological activities of coumarins]. AB - This paper reviewed the advances on effective constituents and biological activities of coumarins in recent ten years. Coumarins are a group of important natural compounds, and have been found to have multi-biological activities such as anti-HIV, anti-tumor, anti-hypertension, anti-arrhythmia, anti-osteoporosis, assuaging pain, preventing asthma and antisepsis. Therefore, further investigation should emphasize on improving techniques for extraction and separation, searching the effective precursory compound, and synthesizing and screening out courmarin derivatives with high activity and low toxicity. PMID- 15810442 TI - [Progress on the research of prevention and treatment of female climacteric syndrome by traditional Chinese medicine]. AB - Perimenopause syndrome, referred also to climacteric syndrome, results from the changing of relationship among hypothalamus, pituitary and ovary during women's aging process. Those changes take place first in overy, then in hypothalmus and pituitary, which are reflected as the functional changes in endocrinological and central nervous system, accompanied with a series of psychological symptoms. 90% of women with perimenopause syndrome show clinical symptoms. The clinical prevention and treatment of female climacteric syndrome by traditional Chinese medicine: traditional Chinese medicine attributes the various symptoms of female climacteric syndrome to a variety of syndromes of Chinese medicine, e. g. gradual consumption of kidney-Qi, emptiness of the Ren and Chong channels, gradual exhaustion of Tienqui, breakdown of Yin-Yang equilibrium in the body, disorders of Zang-Fu, Qi and blood. The treatment based on syndrome differentiation, modification of prescription according to the symptoms, special prescription and simple recipe and acupuncture were employed in treating Yin-deficiency of the kidney, Yang-deficiency of the kidney, Yin and Yang-deficiency of the kidney, breakdown of the normal physiological coordination between the heart and the kidney, deficiency of liver-Yin and kidney-Yin, insufficiency of both the spleen and the kidney, deficiency of Qi and blood in the heart and spleen, stagnation of the liver-Qi and deficiency of the spleen, stagnation of the liver-Qi, phlegm stagnancy and the upward invasion of heat-phlegm. The traditional Chinese medicinal had a satisfactory global regulating effect on the nervous system and immune- regulating network with multifunction and unique advantage, e.g. its regulating effect on FSH, LH, E2, 5-HT, 5-HIAA, free radical and adtonomic nervous system. Experimental studies have also been conducted, e. g. There were also systematic studies of the simple Chinese drugs' pharmacodynamics and drugs forms of prepared drugs. In the health care of female climacteric syndrome the traditional Chinese medicine has also its own features and advantages. PMID- 15810443 TI - [Effects of seeding and sowing methods on the yields of root tubers of Curcuma longa]. AB - OBJECTIVE: To provide a scientific basis for standardizing the cultivation method for Curcuma longa. METHOD: Plant heights and seeding numbers were sampled periodically, the plot yields were counted a tharvested. RESULT: The effects of seeding method on yields, plant height and number were significant. The effects of different sowing ways on the yields were very little. CONCLUSION: The hole seeding method using mother tuber or blastostyle bearing three to four knots was shown to be the best may, worthy to be widely applied in production. PMID- 15810444 TI - [Studies on strain characteristics of Pseudostellaria heterophylla varieties]. AB - OBJECTIVE: To characterize the different varieties of Pseudostellaria heterophylla during cultivation. METHOD: Using systematic selection in the main productive areas, the techniques of random design, all varieties were observed for 3 years. RESULT: The biological and 425 productive characteristics of P. heterophylla var. macrophylla, P. heterophylla var. Foliolum, and P. heterophylla var. anvense were significantly different (P < 0.01). There were also differences in ecological adaptability, plant characteristics, pollen granule, chromosomes, and isoenzyme of the three cultivars. CONCLUSION: The strain types of P. heterophylla was denominated for the first time. The characteristics and productivity index system of P. heterophylla varieties were determined. PMID- 15810445 TI - [Studies on HPLC fingerprint of the hydrophilic constituents of Valeriana medicinal plants]. AB - OBJECTIVE: To establish the fingerprint of the hydrophilic constituents of Valeriana medicinal plants. METHOD: The HPLC-UV assay was used to establish the fingerprint of the hydrophilic constituents of Valeriana medicinal plants. RESULT: The HPLC fingerprint profiles of the hydrophilic constituents of Valeriana medicinal plants contains 12 common peaks. The relative retention time and the ranges of relative area of the common peaks were determined. CONCLUSION: The fingerprint profile can be used for the identification and quality control of Valeriana medicinal plants. PMID- 15810446 TI - [The study on the treatment of Oviducts Ranae protein with enzyme hydrolysis]. AB - The paper introduced a process to enhance the yield of Oviducts Ranae protein using in vitro enzyme hydrolysis. The treatment process included two steps: (1) a 3 - 4 h of hydrolysis of a 0.025 g x g(-1) concentration of substance, at pH 7 and 60 degrees C, using 4% of papain; and (2) followed with a 6 - 8 h hydrolysis, at pH 2 - 2.5 and 60 degrees C, using 3% of pepsin. This treatment process significantly improved the lyophilized Oviducts Ranae in solubility and fluidity, which is convenient for the relative pharmaceutical preparations. PMID- 15810447 TI - [Study on the encapsulation technique of high purity gamma-linolenic acid, part 1 -Single factor, and two-factor tests for encapsulation process]. AB - OBJECTIVE: To study the effects of all factors during the process of urea encapsulation of gamma-linolenic acid on the purity and yield. METHOD: To observe the material proportions, time, temperature and purity using single-factor and two-factor tests. RESULT: Single-factor test showed that the optimal ratio of all materials (oil, urea and 95% ethanol) was 1:3:8. A 30% purity after single encapsulation process was obtained, at the best temperature range was - 15 degrees C-20 degrees C, for 24 hours. Two-factor test showed that the optimal ratio of oil, urea and ethanol was 1:3:8, where the concentration of ethanol was 90%-95%. The purity reached 90% or higher, with three-time encapsulation process. CONCLUSION: An optimized process was identified where material ratios, encapsulation time, temperature, and ethanol concentrations were determined using single-factor and two-factor tests. PMID- 15810448 TI - [The preparation of the polylacticacid nanoparticles of cucurbitacin and their drug loading]. AB - This paper introduced an experimental study of the preparation of polylacticacid (PLA) nanoparticles of cucurbitacin (CuC) using a precipitation method. The residual acetone, ratio of CuC PLA precipitates, and the relationships between the ratios of two precipitates and drug incorporation rates were measured. It appeared that the nanoparticles with 60% of PLA incorporated with 5.5% of CuC were formed when acetone was injected into the aqueous phase. As the acetone gradually evaporated, drug incorporation/encapsulation continued, with most of CuC (about 70%) formed new crystalline cores and suspended in the form of microcrystals in the medium, resulting a suspension containing both nanoparticles and microcrystals. We also concluded that this system may not necessarily be suitable for all lipophilic drugs to be prepared to PLA nanoparticles with good incorporation rate. The drug incorporation depended on the interactions among drug, PLA, and organic solvents, in addition to the solubility of the drug. PMID- 15810449 TI - [Study on RP-HPLC fingerprint of compound hongcao injection]. AB - OBJECTIVE: To establish the fingerprint detecting standard of Compound Hongcao Injection by reversed phase high performance liquid chromatography. METHOD: The chromatographic conditions were as follows: an Inersil-ODS-3 column was used; the mobile phases (acetontrile and 0.1% phosphoric acid) with gradient elution; the flow rate 1.0 mL x min(-1) and the UV absorbance detection at 300 nm. RESULT: Under the same selected chromatographic conditions, the HPLC fingerprints of raw drugs and their intermediate, finished products were obtained with good separation and correlation according to the technical requirements of fingerprint on Injection of Chinese traditional medicine. CONCLUSION: 17 common peaks as well as their retention times and peak area ratios on HPLC fingerprint can be used as the important parameters of the quality control for Compound Hongcao Injection. PMID- 15810450 TI - [Isolation and indentification of chemical compounds from Drynaria fortunei]. AB - OBJECTIVE: To study the chemical components of Drynaria fortunei. METHOD: The herbal material was extracted with 75% ethanol. The chemical components were isolated by silica gel and sephadex LH-20 column chromatography. The structures were identified on the basis of chemical and spectroscopic data. RESULT: Four compounds were isolated, three of which were identified as (-)-epiafzelechin-3-O beta-D-allopyranoside, (-)-epiafzelechin and beta-sitosterol. CONCLUSION: (-) epiafzelechin-3-O-beta-D-allopyranoside, (-)-epiafzelechin were obtained from D. fortunei for the first time. PMID- 15810451 TI - [Studies on chemical constituents in herb of Polygonum orientale]. AB - OBJECTIVE: To study the chemical constituents of the essential substance from Polygonum oriental. METHOD: Compounds were isolated with silica gel and polyamide chromatography and their structures were determined by spectral analysis and chemical evidence. RESULT: Six compounds were obtained and identified as myricitrin, luteolin, gallic acid, catechin, protocatechuic acid, p hydroxycinnamic acid. CONCLUSION: Six compounds were isolated from P. oriental for the first time. PMID- 15810452 TI - [Quantitative analysis of the nucleosides in Cordyceps sinensis with capillary zone electrophoresis]. AB - OBJECTIVE: To establish a quantitative analysis method for analyzing the nucleosides in Cordyceps sinensis with capillary electrophoresis, and compare the difference between natural and the cultured C. mycelia. METHOD: Capillary zone electrophoresis method was employed to quantitate the adenosine, uridine, guanosine and inosine in C. sinensis, with 0.25 mg x L(-1) boric acid-sodium hydroxide buffer, pH 9.5. The working voltage was 20 kV, the temperature was 25 degrees C, and the detection wavelength was 260 nm. RESULT: With the capillary zone electrophoresis method, the average recovery of the above 4 nucleosides was 98.9%, 95.1%, 97.8% and 98.8% respectively, with the RSD 0.4%, 1.7%, 1.3% and 5.0%. There was no adenosine in natural C. sinensis and no inosine in the cultured C. mycelia detected. CONCLUSION: This method can be used to determine the adenosine, uridine, guanosine and inosine in C. sinensis. The nucleosides in C. sinensis produced from Qinghai province and cultured C. mycelia are obviously different. PMID- 15810453 TI - [The effect of kanglaite injection(KLT) on the proliferation and telomerase activity of rat mesangial cells]. AB - OBJECTIVE: To observe the effect of Kanglaite injection(KLT) on the proliferation and telomerase activity of mesangial cells in rats. METHOD: MTT, telomere repeat amplification protocal (TRAP), ELISA, PAGE and silver-stain were applied to detect the growth rate and telomerase activity of MC after stimulation of KLT and IL-1. RESULT: The growth rate of MC was enhanced by IL-1 stimulation, which was accompanied with a redection of the activity of telomerase. Adversely, the growth rate of MC was reduced by KLT, which was accompanied with an enhancement of activity of telomerase. Moreover, the growth rate of MC and the activity of telomerase were both inhibited by the combinative use of IL-1 and KLT without any influence from the sequence of their administration. CONCLUSION: KLT could inhibit proliferation and telomerase activity of MC with or without pre stimulation with IL-1. KLT might be useful to prevent and treat glomerular nephritis related to MC proliferation. PMID- 15810454 TI - [Role of NF-kappaB in the induction of delayed preconditioning in rat heart by pretreatment with Sini decoction]. AB - OBJECTIVE: The present study was designed to investigate whether Sini decoction (SND) containing several chinese medicinal herbs, could induce delayed preconditioning-like effect in rat heart and the possible mechanism. METHOD: In anesthetized open-chest Sprague Dawley rats, left anterior descending coronary artery (LAD) was occluded for 1 h and reperfused vided into four groups: Sham ( n = 8) group in which the surgical procedure was identical to other groups,but the LAD ligature was not ligated; I/R group (n = 8) in which the LAD of rat hearts was subjected to the index occlusion; IPC/SWOP group (n = 8) in which the LAD was occluded for three cycles of 5-minute episodes of preconditioning ischemia followed by 5-minute episodes of reperfusion 24 hours prior to the index occlusion; SND group (n = 8) in which rats were pretreated with Sini decoction (5 g x kg(-1) d(-1)) for three days, the last treatment was pretreated 24 h before the index occlusion. Myocardial infarct size, the activity of superoxide dismutase( SOD) and the content of malondialdehyde (MDA) in myocardial tissue were measured. The mRNA expression of MnSOD and Cu-ZnSOD were detected by RT-PCR. The expression of P65 subunit of NF-kappaB was determined by immunohistochemistry assay. RESULT: As compared with I/R group, myocardial infarct size and the amount of MDA in myocardial tissue were significantly decreased, the activity of SOD and the mRNA expression of MnSOD were increased, the translocation of NF-kappaB was detected and furthermore the expression of NF-kappaB was significantly elevated in SND group as well as in IPC/SWOP group. The mRNA expression of Cu-ZnSOD was lowered in I/R, SND and IPC/SWOP groups, and there was no significant difference among these groups. CONCLUSION: The results suggest that Sini decoction can induce delayed preconditioning-like effect in rat heart and the activation of NF kappaB may play an important role in it. PMID- 15810455 TI - [Protection of puerarin on the cerebral injury in the rats with acute local ischemia]. AB - OBJECTIVE: To study the protection of puerarin on the cerebral injury in the rats with acute local ischemia. METHOD: Rat was evaluated model of acute local cerebral ischemia was made by ligating middle cerebral artery. The cerebral damage toxylin and eosin((HE). RESULT: The number of died neurons were significantly less in puerarin-treated rats than in the rats with cerebral ischemia (P < 0. 05). Similarly, the cerebral edema were significantly attenuated in the puerarin-treated rats as compared with cerebrally ischemic rats. CONCLUSION: Puerarin can prevent the neuron from damage induced by acute cerebral ischemia. PMID- 15810456 TI - [Effects of Naomaitai capsule on learning and memory and content of Ach in rat model produced by cerebral ischemia-reperfusion]. AB - OBJECTIVE: To observe the effects of Naomaitai capsule on the learning and memory abilities and the content of acetylcholine(Ach) in the brain tissue of the model rat. METHOD: One vascular dementia(VD) model was produced by occlusion of bilateral common carotid arteries in rats with following steps: ischemia 20 minutes-reperfusion 10 minutes-ischemia 20 minutes. The learning and memory abilities were tested by water maze. The other VD model was formed by 4-vessel occlusion-reperfusion injury in rats. The time of right reflect recovery and the content of Ach in the brain tissue were determined. RESULT: Naomaitai capsule can significantly improve the learning and memory abilities of the dementia rats( P < 0.05, P < 0.01), increase the content of Ach (P < 0.05, P < 0.01) and reduce the time of right reflect recovery(P < 0.01). CONCLUSION: Naomaitai capsule can improve the learning and memory abilities by increasing the content of Ach of the VD rats. PMID- 15810457 TI - [Study on pharmacokinetics of emodin in Rhizoma Polygontum Cuspidatum and its compound]. AB - OBJECTIVE: To study the difference in the pharmacokinetics of emodin in Zhiganning capsules and Rhizoma Polygontum Cuspidatum by nonaqueous RP-HPLC. METHOD: The rats were orally administered with the extraction of Rhizoma Polygontum Cuspidatum and Zhiganning capsules. After hydrolysis and extraction, the content of emodin in the plasma is determined by Nonaqueous RP-HPLC. RESULT: The concentration-time profiles of emodin fit two-compartment model. The pharmacokinetics parameters including, t1/2alpha, AUC(0-infinity), CL(s) and C(max) of emodin in the group of Rhizoma Polygontum Cuspidatum were significantly different from these in the group of its compounds. CONCLUSION: There is a significant difference in pharmacokinetics of emodin between zhiganning capsules and the extraction of Rhizoma Polygontum Cuspidatum. PMID- 15810458 TI - [Protective effect of ligusticum chuanxiong phthalides on focai cerebral ischemia in rats and its related mechanism of action]. AB - OBJECTIVE: To study the protective effect of ligusticum chuanxiong phthalides on cerebral ischemia in rats and its related mechanism of action. METHOD: Middle cerebral artery occlusion (MCAO) model, thrombosis formation, platelet aggregation and hemorrheological parameters were measured to evaluate the protective effect of ligusticum chuanxiong phthalides. RESULT: Ligusticum chuanxiong phthalides could markedly decrease the infarct size and behavior deficits score, inhibit the thrombus formation and platelet aggregation, ameliorate hemorrheological parameters with a dose-dependent manner in rats. CONCLUSION: Ligusticum chuanxiong phthalides has protective effects on focal cerebral ischemia in rats, and its mechanism may be relevant to its inhibition of platelet-dependent thrombosis and amelioration of hemorrheological parameters. PMID- 15810459 TI - Fundamental steps in experimental design for animal studies. AB - Animal studies continue to have a vital role in science development. The aim of this review is to provide to new investigators an overview of the important steps involved in experimental designs and also to suggest some practical information that is commonly associated with this process. Investigators should adhere to the ethical procedure and follow strictly the scientific method. Both the aims and well-formulated hypothesis are essential and practical. In this regard a profound literature search and the aid of an experienced statistician is encouraged. The need of randomization, blinding, and attempting to minimize variation is discussed and recommended. The choosing of good control groups and the employment of pilot studies are useful. Finally, the formulation of new questions to be further responded is expected. PMID- 15810460 TI - Effect of splenectomy and ligature of the left gastric vein on portal hypertensive colopathy in carriers of surgical hepatosplenic schistosomiasis mansoni. AB - PURPOSE: Esophageal variceal sclerotherapy and band ligation seem not to affect the endoscopic findings of Portal Hypertensive Colopathy (PHC) of cirrhotic patients. The aim was to assess the effect of splenectomy and ligature of the left gastric vein on the PHC in carriers of hepatosplenic schistosomiasis mansoni who underwent this surgery when they were between 9 and 18 year-old. METHODS: Fourteen patients, mean age of 19.1 +/- 3.1 years, were included in the postoperative group (GI). The follow-up was from 1 to 9 years. The preoperative group (GII) consisted of nine patients, mean age of 14.0 +/- 3.1 years. Full length colonoscopy was carried out in all patients. Search was made for PHC lesions. RESULTS: Telangiectasy (GI 100% vs GII 100%), increased vascularisation (GI 57.1% vs GII 100%), focal and diffuse hyperemia (GI 14.3% vs GII 66.7%), angiodysplasia (GI 7.1% vs GII 33.3%), and rectal varix (GI 0% vs GII 55.6%) were the most frequent findings. It was observed that the patients of this series tended to exhibit fewer hemodynamic manifestations of the PHC after treatment (postoperative versus preoperative - chi2 = 8.155 - p = 0.004). CONCLUSION: Splenectomy and ligature of the left gastric vein tend to reduce the abnormal vascular findings of PHC in carriers of hepatosplenic schistosomiasis mansoni. PMID- 15810461 TI - Effects of sodium hyaluronate and carboxymethylcellulose membrane on collagen and fibroblast formation in bowel suture healing: experimental study in rats. AB - PURPOSE: To analyze the effects of sodium hyaluronate and carboxymethylcellulose membrane on collagen and fibroblast formation in bowel suture healing in rats. METHODS: 48 male Wistar rats, weighing 250 to 343g, were randomized into two groups: group I--bowel suture without applying a biologically absorbable membrane and group II--bowel suture with application of an absorbable membrane. The two groups were divided into subgroups of 3, 14 and 30 days of observation, with 8 rats in each subgroup. All were sacrificed after the end of the observation period. RESULTS: No morbidity or mortality was observed during the experiment. The amounts of collagen in group I were 23.4%, 72.1% and 67.6% and in group II were 22.5%, 52.5% and 51.6%, for the subgroups of 3, 14 and 30 days, respectively. Comparison between groups showed that the 14-day (p = 0.0013) and 30-day (p = 0.0587) subgroups had significant variance, with larger collagen zones in animals in which the membrane was not applied. However, with regard to fibroblasts, group I had 2%, 13% and 8% and group II had 2%, 10% and 8%, for the 3-day (p = 1.0), 14-day (p = 0.3184) and 30-day (p = 0.5995) subgroups, respectively, showing no significant variance. CONCLUSION: The use of the biologically absorbable membrane cause a decrease in collagen formation, while not altering the number of fibroblasts, in bowel suture healing in rats, without increased morbidity and mortality. PMID- 15810462 TI - Investigating the relationship between hemodynamic and metabolic parameters in portal triade occlusion following hemorrhagic shock. AB - PURPOSE: To determine whether metabolic and hemodynamic parameters are "mathematically" coupled in rats submitted to portal triad occlusion following controlled hemorrhagic shock state. METHODS: Pearson's coefficient (r-value) analysis was performed. Differences considered significant at p < 0.05 and -0.50 > r > 0.50. RESULTS: It was observed that there is a direct proportional relationship to HCO3- with pCO2 (r = 0.66), base deficit (r = 0.87) and inverse with serum lactate (r = -0.54). pCO2 was directly associated with MAP (r = 0.51), and inversely with pH (r = -0.64). Hematocrit was directly associated with HR (r = 0.72) and CI (r = 0.76), and serum lactated was inversely associated with base deficit (r = -0.61). CONCLUSION: In rats submitted to Pringle's maneuver during 15 minutes following hemorrhagic shock state, there is a mathematical coupling with a very good correlation between several hemodynamic and metabolic variables. PMID- 15810463 TI - [Treatment of complex duodenal lesions: comparison between simple suture and suture with pyloric exclusion and gastrojejunostomy in dogs]. AB - PURPOSE: The goal of this study is to compare the results of two different procedures to complex duodenal lesion (grade III), in an animal model. METHODS: Twenty-four mongrel dogs, weighting 10 to 15 kg, were distributed in 4 groups of 6 animals each. All animals were submitted to a complex duodenal lesion (grade III), with a 50% loss of its circumference. All animals were treated with a longitudinal repair, resulting a significant narrowing of the duodenal lumen. Groups A and C animals were submitted solely to repair while groups B and D, also underwent pyloric exclusion and gastrojejunostomy as a protection method. Groups A and B animals were sacrificed at day 7 post op, while groups C and D were sacrificed at day 14. The following parameters were studied: weight-loss, degree of duodenal stenosis, operative site, vomiting, anastomotic leak, intra-abdominal abscess formation and death. RESULTS: The results obtained with simple duodenal repair were superior to pyloric exclusion and gastrojejunostomy in that the animals lost less weight and vomited less. It was also a simpler and less traumatic procedure. There were no differences in duodenal stenosis, leak, intra abdominal abscess incidence or death between the two treatment groups. CONCLUSION: Duodenal suture associated to pyloric exclusion and gastrojejunostomy resulted more weight-loss and more vomiting incidence; the healing process of the duodenal suture line were similar between the two treatments; both treatments resulted similar increase in the duodenal stenosis degree; there were no cases of suture dehiscence, anastomotic leak, intra-abdominal abscess or death related to the kind of treatment; all animals submitted to pyloric occlusion had it intact at the time of necropsis. PMID- 15810464 TI - [The algimetry evaluation by thermic and pressoric nociceptive stimulus in dogs pre treated with methotrimeprazine, midazolam and ketamine with or without butorphanol or buprenorphine]. AB - PURPOSE: This study aimed at quantifies the pain in dogs under dissociative anesthesia, across thermal and pressoric stimulus and quantify the reasonable period between two different opioids analgesics. METHODS: In this study, 30 dogs were used and, divided into three groups of 10 animals each, in which the animals of GI received methotrimeprazine and midazolam put on the same syringe with ketamine. The animals of GII received the same treatment of GI but associated with butorphanol and finally the animals of GIII received the same treatment of GI but associated with buprenorphine. The routine parametric evaluations has been proceeded, although using the thermo algimetry measured in degrees C with the average of 52 degrees C and the pressoric algimetry in Kg. RESULTS: In the thermo algimetry, there has been significant difference in GI at the moments M0, M1, M4 and M5; in GII it was found at M0, M1, M5 and M6 and in GIII it was observed the significant at M0 and M1. It has also been shown in pressoric algimetry significant difference in GI at the moments M0, M2 and M3. Among GII it has observed significant difference at all moments and it has found at M0, M9 in GIII. Thus, it has observed significant differences between all groups; for such the M2 of GII smaller than the others; and M4, M5 of GIII bigger than GI and GII. In the assessment of all periods it was observed significant latent period bigger in GI, however, with reasonable period and short recovery in GII and GIII. In the order hand, the postural tonus recovery it was longer in GIII, followed by GII and finally GI. CONCLUSION: The used method for the measurement of algic stimulus was efficient, noticing a reasonable analgesic period of 3 hours for butorphanol and 6 hours for buprenorphine. PMID- 15810465 TI - [Comparative study among five different treatments on the clinical and laboratory changes in rats with alloxan-induced diabetes]. AB - PURPOSE: The long-term effects of five different treatments of diabetes were evaluated in alloxan-induced diabetic rats. METHODS: Seven experimental groups, with 50 rats each (GN--normal control; GD--untreated diabetic control; GI, GA, GIA--treated groups with insulin, acarbose, and insulin plus acarbose, respectively; GTIL, GTPD--treated groups with islet of Langerhans and pancreas transplantation) were studied. Clinical (body weight, water intake, food intake and urine output) and laboratory (blood and urinary glucose, and plasma insulin) parameters were analyzed at the beginning of the study, and after 1, 3, 6, 9 and 12 months of follow-up. RESULTS: Mortality was observed in all groups, except GN, during 12 months (GD = 50%; GI = 20%; GA = 26%; GIA = 18%; GTIL = 4%; GTPD = 20%). Rats from the GD, GI, and GIA groups died due to metabolic or hydrossaline disbalance, and/or pneumonia, diarrhoea, and cachexy. All deaths observed in GTIL and GTPD groups were in decorrence of technical failure at the immediate postoperative, until 72h. Animals from the GI, GA and GIA had significative improving of the clinical and laboratory parameters (p < 0,05) observed in diabetic rats, being the efficacy of theses treatments equal. However, rats from the GTIL and GTPD groups had better control of these parameters than GI, GA, and GIA groups. Transplanted rats had complete restoration, at the normal levels, of all analyzed variables (p < 0.01). CONCLUSIONS: Conventional treatments with insulin, acarbose, and insulin plus acarbose improved the severe diabetic state of the alloxan-diabetic rats, but pancreas and islet transplantation have a better performance for treatment of diabetes. PMID- 15810466 TI - [Ileocecal valve reconstruction in dogs]. AB - PURPOSE: The importance of keeping the ileocecal valve in the intestinal ressections has been reported by several authors. When preserved, the ileocecal valve was related to a longer survival and prevention of the short bowel syndrome, due to its ability to block the colonic content reflux into the ileum and to avoid the rapid empting of the ileal content into the cecum. It was assessed a tecnique of ileocecal valve reconstitution, based on vesicoureteral anti-reflux tecniques. METHODS: Fourteen beagles were operated. Seven underwent ileocecal valve reconstitution following the tecnique proposed and in the other seven a simple end-to-end anastomosis was performed. To assess the new valve, it was done the clinical follow up, the microbiologic analysis and the manometric study. RESULTS: Clinically, during 45 days of follow up, there was no difference between the dogs with and without ileocecal reconstitution. In the aerobic bacteria analysis, the predominant bacterium was Escherichia coli. Quantitatively, the cultures grew in an irregular way, so that it was not able to compare the bacterial growth between the groups with or without ileocecal valve. The new valve had a colo-ileal reflux pressure similar to that of the physiological valve (P > 0.05). However, when compared to the non valve group, the reflux pressures of the physiological valve and new valve were significantly higher, with P < 0.05 and P < 0.001, respectively. CONCLUSION: In this study, the reconstituted ileocecal valve served as a barrier to the colo-ileal reflux just as the physiological valve does. PMID- 15810467 TI - [Model of inflammatory angiogenesis in rabbit cornea induced by punctual alkaline cauterization]. AB - PURPOSE: To establish a model of angiogenesis in rabbit cornea induced by punctual alkaline cauterization. METHODS: Six rabbits were submitted to punctual cauterization in right cornea. It was used a circular piece (3 mm) of filter paper, that was previously soaked in 1 M NaOH and placed 1 mm from the superior limbus for 2 minutes. The animals were evaluated at 3, 6, 9, 12, 15, 18 and 21 days after cauterization. Quantification of angiogenesis was performed according to 4 methods: radial vessel length (V), angiogenic index (I), area of the triangle (T) and area of the circular band sector (S), which provide an approximation of the area of neovascularization. RESULTS: The analysis of the quantitative data of angiogenesis showed that neovascular response progressed in a biphasic manner: rising between days 0 and 12 (proliferation), stable between days 12 and 21 (maturation). This pattern was observed on the values of the vascular growth mean rate. Between days 0 and 12, it was 16 times greater than that verified between days 12 and 21. It was found a statiscally significant positive linear correlation among the 4 methods of measurement of angiogenesis. CONCLUSION: The punctual alkaline cauterization of the rabbit cornea induces and sustains corneal neovascularization, so that, it can be used as model of angiogenesis. PMID- 15810468 TI - [One layer end to side choledochusduodenal anastomosis with evertion of the choledochus wall in swines]. AB - PURPOSE: This article aims to study the one layer end to side choledochusduodenal anastomosis creating a "neopapila" which, because of the mucosa recovering it, would prevent stenosis. METHODS: Thirty male pigs were distributed within 3 equal groups = Group I (control)--conventional choledochusduodenal anastomosis. Group II (experiment)--one layer end to side anastomosis with evertion of the choledochus wall (neopapila). Group III (sham)--isolation and mobilization of the biliar tract with no choledocotomy. We got the biliar flow and pressure (pre and postoperative) monitorized on the 15 PO day the animals went trough an endoscopy and on the 30 PO day they were operated and the tissues were analysed under macro and microscopic view. RESULTS: Regarding to biliar flow and manometry there were differences between all groups but the III. Animals in groups I and II showed intestinal metaplasia and diminished elastic fibers in the choledochus. All animals showed choledoco dilation. CONCLUSION: The proposed technique allowed the same results as the standard technique. PMID- 15810469 TI - [Peritoneal-fascia-muscle autologous graft in the inguinal canal of rats]. AB - PURPOSE: To study the fragment of the autologous tissue, built by a peritoneum fascia-muscle, implanted on the level of the wall of inguinal canal and the spermatic cord of rats aiming to verify the histological behavior of the survive and the tissue incorporation. METHODS: Thirty-six rats were distributed into four groups. The rats of the group A, B, C and D were submitted to the median laparotomy to obtain the species to be implanted. On the group A, was implanted a tissue compounded by peritoneum and transversal fascia on the wall of the inguinal canal. On the B, the tissue implanted was added a part of rectum muscle of the abdomen and implanted on the wall of the inguinal canal. On the C, the tissue grafted was similar to the groupA, now grafted into the spermatic cord. On the D, the tissue was similar to the group B, now grafted into the spermatic cord. RESULTS: After 21 days of implant, these tissues presented the following histological alteration: the group A and C, fibroadipous tissue with dilated vessel, Focus of fibrosis' and limphoplasmocitary infiltrated and the groups B and D, fibroadipous tissue with dilated vessels, skeletal muscle and focus of fibrosis. The fibrosis displayed on the groups B and D was more intensive. On the group B, we almost always find the skeletal muscle, while on the group D this tissue was presented in six animals, almost replaced by fibrosis in two and not present in one. CONCLUSION: All the implanted grafts displayed survive and local incorporation, what could be useful on the correction of defects on the inguinal canal, on situations when the autologous tissue was indicated. PMID- 15810471 TI - [Prevention of median incisional hernia in rats by ultrasonic therapy]. AB - PURPOSE: To investigate if ultrasonic therapy is effective in avoidance of incisional hernia (IH), once the ideal technique for repair of IH is still a non solved problem, in despite of prostheses usage. METHODS: IH was created by linea alba opening over 20 male Wistar rats, distributed in two groups: one group received the ultrasound, over the wound, during 14 days (5 animals) and the other for 28 days (5 animals), each one with the same number of animals as controls. The ultrasound was applied using a small probe, 1:5 pulse mode, 3.0 MHz frequency and 0.5W/cm2 intensity, for 5 minutes daily, from the first postoperative day. The width of the linea alba scar or the hernia ring was measured at the end of every period. The IH was defined when these openings were over 2mm wide. Some specimens from hernia sac or linea alba scar were obtained for vessels, fibroblasts, and colagen fibers density. Standard deviation and medians were obtained by variation analysis and Tukey-Kramer multi-comparison test. RESULTS: The ultrasound group showed a linea alba scar in 60%. The other 40% presented the hernia ring smaller than the non treated group (p < 0.0001). The ultrasound increased density of blood vessels (p = 0.0031), fibroblasts (p < 0.0001), and collagen fibers (p = 0.0015). There was no difference between the two treated groups. CONCLUSION: The ultrasound avoided the IH to appear or reduced its width. Fourteen days of treatment were sufficient for this purpose. PMID- 15810470 TI - [Copaiba oil effect on aminotransferases of rats with hepatic ischemia and reperfusion with and without ischemic preconditioning]. AB - PURPOSE: To study the copaiba oil effect in rats' aminotrasnferases submitted to hepatic ischemic and reperfusion with and without preconditioning. METHODS: 24 male and adults rats (Rattus norvegicus albinus, Wistar) were distributed into six groups: Standard Group (SG); Copaiba Group (CG), Ischemic-reperfusion Group (IRG), Ischemic-reperfusion + Copaiba Group (IRCG), Ischemic Preconditioning Group (IPCG) and Ischemic Preconditioning + Copaiba Group (IPCCG). The animals of CG, IRCG and IPCCG received copaiba oil, 0.63 ml/kg/day by gavage, during 7 days. It was realized total hepatic ischemic by 30 minutes and, in animals subbimited to ischemic preconditioning, it was realized 10 minutes of ischemia, following by 5 minutes of reperfusion and other ischemia by 30 minutes. All animals were subbmited to 24 hours of reperfusion. To realize surgery procedure, animals were anesthezied through ethilic eter breathing. On first post-operating day, blood was collected to dose aminotransferases. RESULTS: It was not observe alterations in AST level in animals which received copaiba oil. There was no statistic difference between IRCG and IRG ALT levels, however, this enzyme was increased in IPCCG when compared with IPCG. CONCLUSION: Copaiba oil did not change AST levels in groups studied. Analysing ALT, this oil did not change its values in animals were realized just hepatic ischemic-reperfusion, however, it had levels increase in IPCCG when compared to IPCG Between IRCG and IPCCG, ALT levels were not statistical different. PMID- 15810472 TI - [Anal cytology as a screening method for early detection of anal cancer: are hydrophilic cotton smears really unsatisfactory?]. AB - PURPOSE: To test the ability of analpaps performed with cotton swab smears to produce satisfactory cytologic readings as compared to traditional sampling materials. METHODS: 318 consecutive patients were examined at the Ambulatorio Araujo Lima of Hospital Universitario Getulio Vargas in the Anal Cancer Prevention Week and were sampled for the performance of analpap. 180 of these patients were randomly sorted to be sampled by either one of the following materials: cytologic brush, dacron swab or cotton swab. The ability of cotton in producing satisfactory anal cytologic readings as compared to dacron and cytologic brush was analised. RESULTS: There was no statistical difference among the materials employed in the ability to produce satisfactory citological readings in analpaps. (p > 0.05) CONCLUSION: Our results showed that cotton swabs were as efficient as dacron swabs or cytologic brushes in the ability to produce satisfactory analpap readings. PMID- 15810473 TI - Plaque removal efficacy and safety of the next generation of manual toothbrush with angled bristle technology: results from three comparative clinical studies. AB - PURPOSE: To investigate the safety and efficacy of a novel angled-bristled toothbrush in comparison with three established brushes. METHODS: The Oral-B CrossAction Vitalizer toothbrush was compared with two manual toothbrushes, the Oral-B CrossAction and Oral-B Advantage, and the battery-operated Crest SpinBrush Pro brush in three independent single-use, examiner-blind, crossover studies. In each study, over 50 healthy subjects from a normal population brushed with their randomly assigned toothbrush for 1 minute without instruction. Subjects returned after a 1-week washout period and brushed with the alternate toothbrush. At each visit, oral hard and soft tissues and plaque were examined before and after brushing. Plaque was evaluated using the Rustogi Modified Navy Plaque Index. RESULTS: Each tested toothbrush significantly (P=0.0001) reduced plaque levels after a single brushing. However, in all three studies, the CrossAction Vitalizer was significantly (P=0.0001) more effective than the comparator brushes in plaque removal from the whole mouth, the gingival margin and approximal surfaces. All toothbrushes were found to be safe with no evidence of oral hard or soft tissue trauma. PMID- 15810474 TI - Subsurface microhardness of enamel and dentin after different external bleaching procedures. AB - PURPOSE: To evaluate the influence of various bleaching systems on subsurface microhardness of enamel and dentin. METHODS: 60 bovine crowns were distributed among seven groups (A: Opalescence Xtra Boost, B: Opalescence Quick, C: Rapid White, D: Whitestrips, E: Opalescence 10%, F: Opalescence PF 15%). The crowns were sectioned and baseline hardness (Knoop) of enamel and dentin was assessed on the sectioned surface at various distances from the enamel surface. The sectioned surface was covered with wax and the enamel was treated for 10 days with the bleaching agents A-F according to manufacturers' instructions. Bleaching with C-F was conducted each day (C: twice per day for 10 minutes, D: twice per day for 30 minutes, E: 8 hours, F: 4 hours), systems A-B were applied on first and fifth days (A: twice for 10 minutes, B: 1 hour). Finally, hardness was re-assessed. Data were statistically analyzed to compare baseline hardness and final hardness in the respective groups (P< 0.05). RESULTS: Analysis showed that in Group C, hardness was significantly reduced in both enamel and dentin. In the remaining groups, significant reduction of hardness was observed up to the following depths [microm] in enamel; A: 250, B: 700, D: 300, E: 150, F: 150. In these groups no significant hardness changes were recorded in subsurface dentin. PMID- 15810475 TI - Peroxide release into saliva from five different home bleaching systems in vivo. AB - PURPOSE: This study determined hydrogen peroxide release into the oral cavity during use of different home bleaching products and compared them with accepted safe levels. METHODS: Determination of peroxide in saliva was performed with peroxidase, phenol and 4-aminoantipyrin in a photometric method. Upper jaw incisors were bleached with individual trays charged with 350 mg Opalescence 10%, Opalescence 15% (OP) and Vivastyle (V). Additionally, Whitestrips (WS) designed for upper or lower jaw were used. All systems were adopted by five subjects for 30 minutes on different days. Whole saliva was collected at 2-minute intervals during the first 10 minutes of bleaching and every 5 minutes thereafter. RESULTS: Highest release of peroxide was found for all products in the saliva sample collected initially after application of the bleaching agent. Total amount of peroxide released into saliva during 30-minute bleaching period was 0.78+/-0.45 for Opalescence 10% and 1.52+/-0.44 mg for Opalescence 15%. Significantly more peroxide was released from Vivastyle (2.67+/-1.03 mg) and from Whitestrips (upper: 3.25+/-5,65, lower: 2.09+/-0.34 mg). A significantly smaller fraction of the charged peroxides was released into saliva from individual trays than from Whitestrips during the 30-minute use time. From the peroxide loaded in the trays or strips the following fractions were released during the application period: Opalescence 10% (6.4+/-3.7%), Opalescence 15% (8+/-2.4%), Vivastyle (18.6+/ 8.5%), upper Whitestrips (30.4+/-4.9), lower Whitestrips (27.4+/-4.4%). In terms of amount/kg body weight the bleaching systems led to a single exposure of 0.013 0.056 mg/kg which is distinctly less than the maximum safe daily dose of 0.26 mg/kg/day if calculated for a small person (58 kg/128 lbs). PMID- 15810476 TI - Longevity of bonds made by composite and polyacid-modified resins to dentin using a dual-cured adhesive system. AB - PURPOSE: To evaluate the microtensile bond strengths (MTBS) of a dual-cured dentin adhesive bonded to dentin using a light-cured resin-based composite (composite) or a polyacid-modified resin-based composite (compomer), and to investigate the effect of water aging on the bond strengths of these systems. METHODS: Flat dentin surfaces prepared from extracted human third molars were bonded with Prime&Bond NT Dual-Cured version (PBNT). Resin build-up crowns were constructed incrementally with two different materials: a composite (Z250) or a compomer (Dyract AP). When Dyract AP was used as the restorative material, half of the specimens were etched with 36% phosphoric acid, and the other half of the specimens were not etched. After storage in water for 24 hours or for 6 months, the teeth were sectioned to obtain bonded 1.3 mm2 beams containing the bonded interface. Each beam was tested in tension in an Instron machine at 0.5 mm/minute. Results were analyzed by multiple ANOVA and Student-Newman-Keuls multiple comparison tests. RESULTS: The group PBNT/Dyract AP bonded to acid etched dentin showed the highest bond strength, in the 24-hour evaluation. Bond strength values decreased significantly after aging. The groups treated with PBNT in light-cured, dual-cured and self-cured modes performed equally well in the 24 hour evaluation. After 6 months of water aging, bond-strengths of the light-cured groups remained stable. By contrast, decreases in the bond strengths of the self cured groups were observed. PMID- 15810477 TI - Evaluation of resin composite materials. Part I: in vitro investigations. AB - PURPOSE: To evaluate different resin-based composites using a variety of in vitro investigation methods to predict their clinical behavior. METHODS: Materials selected for this study were Heliomolar radiopaque (microfilled), Tetric Ceram, Pertac II (minifilled hybrids), Ariston pHc (ion releasing hybrid), and Solitaire I (hybrid with porous fillers). The evaluated in vitro criteria were three-body wear according to the ACTA method, microtensile bond strengths to enamel and dentin, flexural strength (four-point bending strength), flexural fatigue behavior (flexural fatigue limit), and calcium ion release (for Ariston pHc). RESULTS: Concerning wear resistance, Ariston pHc (20.5 microm after 200,000 cycles) was inferior to the other materials (13.7-15.9 microm). Microtensile bond strengths to dentin were similar for Heliomolar (32.0 MPa), Tetric Ceram (30.4 MPa; both bonded with Syntac Classic), and Pertac II (30.8 MPa; bonded with EBS Multi) being above Solitaire I (22.5 MPa; bonded with Solidbond) being above Ariston pHc (13.2 MPa; bonded with Ariston Liner). Enamel bond strengths for Heliomolar (40.0 MPa), Tetric Ceram (36.5 MPa), and Pertac II (38.9 MPa) were significantly higher than for Solitaire I (26.6 MPa) which was above Ariston pHc (7.2 MPa). Heliomolar, Tetric Ceram, and Pertac II revealed higher micro-TBS to enamel than to dentin, Ariston showed the contrary, and Solitare exhibited no difference between enamel and dentin micro-TBS. Solitaire I exhibited a lower initial flexural strength than the other materials, the computed fatigue strength of the material even dropped to the level of glass ionomer cements (17.9 MPa). Long-term calcium release data for Ariston exhibited a continuously high calcium release becoming lower at the end of the observation beyond 21 months. PMID- 15810478 TI - Pit and fissure sealing. Bonding of bulk-cured, low-filled, light-curing resins to bacteria-contaminated uncut enamel in high c-factor cavities. AB - PURPOSE: To examine the ultrastructure of bonding to occlusal enamel fissures using phosphoric acid etching in combination with a pit and fissure sealant (Clinpro Sealant), a total-etch single-bottle adhesive (One-Step) followed by the fissure sealant, and two single-step self-etch adhesives (Adper Prompt and Xeno III) followed by the fissure sealant. METHODS: Sections of bonded enamel fissures retrieved from specimens bonded in vivo were polished, and rinsed with phosphoric acid to bring surfaces into relief. They were examined, first under non dehydrated conditions with field emission-environmental SEM, to evaluate the marginal integrity of the bonds. The same specimens were then re-examined under dehydrated conditions with conventional SEM to capture more detailed ultrastructural features. Extent of resin penetration into etched enamel was further evaluated by TEM examination of sections taken from undemineralized unstained bonded specimens. These sections were further examined after HCl demineralization and staining. RESULTS: All occlusal fissure walls examined were lined with remnant aprismatic enamel. Full penetration of resins into the bottom of the fissures, and gap-free interfaces were not observed in any of the specimens, irrespective of whether total-etch or self-etch adhesives were used with the sealant. Phosphoric acid did not penetrate well into the fissures and although hybridization of the etched aprismatic enamel was observed with or without the use of the total-etch adhesive, etching was inconsistent and gaps were frequently observed. Entrapment of bacteria within the fissure walls was frequently seen. The more aggressive self-etch adhesive Adper Prompt created etching in aprismatic enamel that approached that of phosphoric acid etching. The moderately aggressive self-etch adhesive Xeno III produced 1 mm thick hybrid layers in the aprismatic enamel fissure wall. PMID- 15810479 TI - Microtensile bond strength of total-etch and self-etch adhesives to the enamel walls of Class V cavities. AB - PURPOSE: To determine the difference in microtensile bond strengths (MTBS) of resin-based composite bonded to the enamel of cavity preparations using a conventional or a self-etching adhesive. METHODS: Buccal and lingual Class V cavities were prepared in each of 12 caries-free human molars. Six teeth were randomly selected to be restored with Spectrum TPH resin-based composite and Prime & Bond NT adhesive. The remaining six teeth were restored with the same composite and Prompt L-Pop self-etching adhesive. Enamel/composite sticks were cut from each specimen and subjected to microtensile bond testing. Each debonded specimen was evaluated by SEM to determine the mode of failure. RESULTS: The mean MTBS for Prime & Bond NT specimens was 15.2+/-13.1 MPa. The mean MTBS for Prompt L-Pop self-etching adhesive samples was 18.2+/-14.3 MPa. ANOVA demonstrated no statistically significant difference between the mean MTBS associated with the two adhesives (P> 0.05). The results of SEM analysis revealed that the bonding failures occurred primarily at the interface between the adhesive and the enamel, suggesting a primarily adhesive failure. PMID- 15810480 TI - Effect of application mode on bonding performance of self-etching adhesives. AB - PURPOSE: To investigate effects of static vs. dynamic application of self-etching adhesives on bond strengths to enamel and dentin and marginal adaptation in dentin cavities. METHODS: Products: AC Bond experimental (ACB), AQ Bond (AQB), Fluorobond (FLB), and One-Up Bond F (OUB). Shear bond strengths (SBS) to enamel and dentin were determined on human molars. Primer applications: 1. undisturbed, 2. brush agitation, 3. ultrasonic agitation (Cavitron, lowest setting), or 4. ultrasonic agitation (5 seconds). A 3.5 x 2 mm mold was mounted for application of the resin-based composite Charisma. Six specimens for each condition were stored for 24 hours and loaded until failure. STATISTICS: ANOVA and Scheffe test (P= 0.05). Marginal adaptation was determined in cylindrical dentin cavities (4 x 1.5 mm) 15 minutes after curing of Charisma. Eight specimens/adhesive were produced following application modes 1, 2, and 4. Kruskal-Wallis and Mann-Whitney showed a P= 0.05. RESULTS: Application modes showed no differences in shear bond strengths (SBS) on enamel and dentin by adhesive. SBSs on enamel were not different from those on dentin. Average SBSs for ACB, AQB, FLB, and OUB were 21.0, 21.9, 20.8, and 13.7 MPa. Ranking of adhesives for SBS on enamel and dentin was: AQB, ACB > ACB, FLB > OUB. Application modes had no influence on marginal adaptation. From the 24 cavities each, all ACB, 21 AQB, 13 FLB, and 1 OUB were gap-free. Mann-Whitney ranking: ACB, AQB, FLB > OUB. PMID- 15810481 TI - Four-year clinical evaluation of a self-etching primer and resin-based restorative material. AB - PURPOSE: To evaluate the performance of a universal resin-based restorative material (Beautifil) in combination with a self-etching primer (Fluorobond) for posterior restorations. METHODS: 61 restorations, 26 Class I and 35 Class II, were placed by two clinicians in 31 patients under rubber dam isolation. Two other calibrated clinicians evaluated the restorations using USPHS/Ryge criteria observing the following characteristics: color match, marginal adaptation, anatomy, roughness, marginal staining, interfacial staining, contact, secondary caries, and luster. Chi-Square and Fisher's Exact Test were used for statistical analysis (alpha=0.05). RESULTS: All the restorations were assessed at baseline and alpha scores predominated for all criteria. All restorations were examined at the 1-year recall examination. 58 restorations were examined at the 2-year recall (23 Class I and 35 Class II), 47 (16 Class I and 31 Class II) at the 3-year recall. The present 4-year report covers 39 restorations (12 Class I and 27 Class II). Marginal adaptation, staining, anatomy, and color matching constitute the majority of the recorded changes. Chi-Square and Fisher's Exact Test of the frequency of clinical rating of each characteristic showed that there were no statistically significant differences at each recall with respect to the baseline observations (P> 0.05), except for occlusal marginal stain. Statistical significance was detected for the occlusal marginal stain at 1-year (P= 0.06), 2 year (P= 0.01) and 4-year (P= 0.002) when compared to baseline data. PMID- 15810482 TI - Accuracy of assessing infrabony defects using a special digital filter for periodontal bone loss. AB - PURPOSE: To assess the accuracy of linear measurements of bone loss within infrabony defects on digitized radiographs after digital filtering particularly designed to image periodontal bone loss. METHODS: Before surgery, 50 standardized radiographs of 50 infrabony defects were obtained. Intrasurgically the distances from the cementoenamel-junction (CEJ) to the alveolar crest (AC), from the CEJ to the bottom of the bony defect (BD), and the depth of the infrabony component (INFRA) were assessed. All radiographs were digitized and the distances CEJ-AC, CEJ-BD, and INFRA were measured on digitized but unchanged radiographs and after use of the filter. RESULTS: There was a small overestimation of CEJ-AC (0.35-0.68 mm) and underestimation of CEJ-BD (0.84-1.12 mm) and INFRA (0.66-0.76 mm). Accuracy of measurement of the distance CEJ-AC was influenced by vertical angulation difference (P= 0.047) and the intrasurgical CEJ-AC (P= 0.003). Accuracy of CEJ-BD was influenced by intrasurgically assessed bone loss (P= 0.012). Accuracy of INFRA was influenced by individual patient, intrasurgically assessed INFRA (P< 0.001), and filter (P= 0.018). PMID- 15810483 TI - Radiodensity evaluation of seven root post systems. AB - PURPOSE: To compare through digital radiographic analysis the radiodensity levels of seven prefabricated post systems inserted in root canals of bovine extracted incisors. METHODS: 21 bovine teeth were cut 15 mm from the apical limit. The canals were prepared according to the manufacturer's instructions using specific bur of each post system analyzed: three metallic posts: Flexi-Flange, Radix Anker, and FKG post; one zirconium ceramic post: Cosmopost; one carbon fiber post: C-Post; one carbon fiber post system covered with quartz fiber: AEsthetic Post; and one glass fiber reinforced post: FibreKor Post. Root tooth digital analyses were made with Digora system with and without the insertion of post. Their variations were obtained subtracting radiodensity of the post from radiodensity of the root canal, in cervical, medium, and apical thirds of the root canal. Two-way ANOVA and Tukey test (P< 0.05) were used to analyze the data. RESULTS: This study showed that the radiopacity of post systems is dependent on their composition. Statistical differences were found among radiodensity levels in apical third, medium and cervical thirds for all post systems inside the root canals. Statistical differences were also found among the post systems: ceramic posts showed the highest radiodensity level, followed by metallic posts, carbon fiber posts, glass fiber posts, and the carbon fiber post covered with quartz fiber. These results indicate that biomechanical post systems have low radiodensity levels. PMID- 15810484 TI - Scanning electron microscopic evaluation of fiber post-resin core units built up with different resin composites. AB - PURPOSE: To microscopically evaluate the structural characteristics of post-and core units made of a fiber post and of different types of resin-based composites used as core materials. METHODS: 55 endodontically treated human maxillary incisors were prepared for receiving a fiber post (AEsthetic Post Plus). One-Step and C&B resin cement were used for luting the post. Samples were randomly divided into 11 groups. Eleven different composite materials were selected to build-up the abutment. The materials on trial were: Group 1: Z100; Group 2: Light-Core; Group 3: Lumiglass; Group 4: Gradia Direct; Group 5: Build-It!; Group 6: Tetric Ceram; Group 7: Biscore self-curing; Group 8: Unifil Flow; Group 9: Definite Flow; Group 10: Tetric Flow; Group 11: AEliteFlo. All the post-and-core specimens were cut perpendicularly to their long axis and processed for SEM observations. The presence of voids/bubbles within the resin abutment and of defects at the interface between the post surface and the core material was scored. These aspects were quantified with reference to a predefined index. The differences among the scores were tested for statistical significance (P< 0.05). RESULTS: Cores built up with flowable composites showed the highest integrity and the best adaptation onto the post. PMID- 15810485 TI - Evaluation of the interface between one-bottle adhesive systems and dentin by Goldner's trichrome. AB - PURPOSE: To evaluate the hybrid layers in interfaces between dentin and six contemporary one-bottle adhesive systems using a nondestructive differential staining micro-technique. METHODS: The adhesive systems used in this study were divided to three groups based on hydrophilic/hydrophobic component ratios (i.e. ability to dissolve in water), from highest to lowest as follows: Group 1 (Dentastic UNO.DUO, PermaQuick PQ1) > Group 2 (One-Step, Primer&Bond NT) > Group 3 (Optibond Solo, Single Bond). The occlusal third of the crown was removed from 36 extracted, unerupted human 3rd molars. Smear layers were created by abrading the dentin with 600 grit SiC under water. The exposed dentin was treated with one of the adhesive systems per manufacturer's instructions. After 24 hours in water, 3-5 microm thin sections of the adhesive/dentin (adhesive/dentin ) interface were cut with a microtome and stained with Goldner's trichrome. Stained thin sections from each prepared tooth were imaged with light microscopy. RESULTS: The thickness and color difference of adhesive/dentin interfaces among these one bottle adhesive systems were clearly visualized. The width of the hybrid layers varied, ranged from 4.1-9.2 microm for six adhesive systems. The color differences in the stained sections are reflected to the extent and degree to which the adhesive envelops the exposed collagen. Among these six bonding systems, resin encapsulation of collagen varied from highest to lowest as follows: Group 1 (UNO, PQ1) > Group 2 (OS, PBNT) > Group 3 (OP, SB). The differences in collagen encapsulation are dependent on the adhesive composition, or the ability to tolerate water during the infiltration of the wet demineralized matrix. PMID- 15810486 TI - William Crowley, MD. PMID- 15810488 TI - Implementation of NIH on-line archive of research publications revised to reflect publishers' objections. PMID- 15810487 TI - Reaction mixed to sweeping NIH ethics regulation. PMID- 15810489 TI - Federal/Private partners launch resource for diabetic kidney disease gene studies. PMID- 15810490 TI - NIH releases Action Plan for liver disease research. PMID- 15810491 TI - Advancing excellence in health care: getting to effectiveness. PMID- 15810492 TI - Creating a new structure for research on health care effectiveness. AB - Effectiveness research (a term we use in preference to the more confining and difficult health services or outcomes research) evaluates the clinical setting and the health care system on which it depends. It uses a variety of health care assessment techniques and the practical clinical trial to inform clinical practice, quality interventions, and health policy decisions. Effectiveness research had not had sufficient public or private funding to produce the information needed to facilitate evidence-based health care improvement. However, recent trends, such as the likelihood for continued substantial increases in health care costs and concern regarding the quality and safety of the US health care system, are among the important arguments for increasing its funding and capacity. We propose a new entity, a public-private consortium to expand and offer new capability and resources in this area. The consortium would consist of all relevant public and private entities. It would be organized into an executive committee, which would identify research priorities and panels to design requests for proposals. Competitive peer-reviewed proposals, transparency and balance of forces in choice of topics, conduct of research, and interpretation of results would be important features. Metrics for success would be use of the data derived from consortium projects in medical decision making and benefit design. The consortium would provide balance and potential mediation of conflicting or competing interests in which all stakeholders will be present to establish the rules. Broad representation of all interests would serve to avoid the economic, policy, and political issues that have bedeviled past efforts. Models for the consortium include the Health Effectiveness Institute, the Centers for Education and Research on Therapeutics, and the Transportation Research Board. PMID- 15810493 TI - Launching a new fellowship for medical students: the first years of the Doris Duke Clinical Research Fellowship Program. AB - As part of its commitment to increasing the pipeline of physicians pursuing careers in clinical research, the Doris Duke Charitable Foundation launched the Doris Duke Clinical Research Fellowship (CRF) Program for medical students in 2000. The program, which is based at 10 US medical schools, provides medical students from any US medical school with the opportunity to spend 1 year obtaining both didactic and "hands-on" mentored clinical research experience. This article describes the program and summarizes the early outcomes collected during the first 3.5 years of the program. Interest in the program among medical students has been robust and widespread, with 35% of CRF program fellows matriculated at non-CRF schools. Exit surveys of the first three classes of fellows totaling 174 fellows indicated that (1) 97% of the fellows felt that participating in the fellowship was a good decision; (2) commitment to a career in clinical research was increased among the 44% of fellows reporting that they were unsure about pursuing a clinical research career when they began their fellowship; (3) there was no difference in satisfaction level between the fellows who remained at the medical schools in which they were matriculated and those who completed their fellowship at a medical school in which they were not matriculated; and (4) the majority of fellows responded that the didactic component of their fellowship was useful. PMID- 15810494 TI - Readaptation from simulated microgravity as a stimulus for improved orthostatic tolerance: role of the renal, cardioendocrine, and cardiovascular systems. AB - BACKGROUND: Microgravity and simulated microgravity (SM) lead to important changes in orthostatic tolerance (OT), the autonomic nervous system (ANS), and the volume-regulating systems. After one is exposed to microgravity or SM, a period of readaptation to gravity is known to take place, but it is not certain if orthostatic function returns to baseline within the initial recovery and what mechanisms are involved. We hypothesized that after a period of recovery, OT, ANS, and volume-regulating systems would return to pre-SM levels. METHODS: To test this hypothesis, 24 healthy men were placed on a constant diet for 3 to 5 days, after which a tilt-stand test (pre-TST) was performed. The TST was repeated after 14 to 16 days of head-down tilt bed rest (HDTB) (post-TST) and a 3-day period of recovery (rec-TST), at which times measurements of renal, cardioendocrine, and cardiovascular systems were conducted. RESULTS: Presyncope occurred in 46% of subjects pre-TST, in 72% post-TST, and in 23% during rec-TST. OT was significantly better during the recovery period than at baseline (p = .03). There was a significant decrease in urinary sodium and potassium excretion, along with a decrease in plasma renin activity and serum and urine aldosterone compared with baseline. Serum norepinephrine and sympathetic responsiveness remained below baseline values. CONCLUSION: In summary, OT improved compared with baseline after a period of readaptation. Retention of electrolytes (sodium, potassium) could be involved. These findings indicate that recovery after SM is not simply a gradual return to baseline values but is instead a dynamic process reflecting interaction of multiple regulatory systems. PMID- 15810495 TI - Exclusive CLTC research: Americans prefer faith-based facilities. PMID- 15810496 TI - Aging in place challenges assistance in IL. PMID- 15810497 TI - Is that what your residents ordered? PMID- 15810498 TI - [Results of the work of sanitary-epidemiologic service in Poland during last 85 years and perspectives for the future]. AB - Just after the First World War, when Poland received independence, activity of sanitary-epidemiological service was included as the most important in the country. In this time epidemiological situation of infectious diseases was unsuccessful, especially concerning typhus exantematicus, typhoid fever, dysentery, diphtheria, tuberculosis. National Central Epidemiological Institute and since 1923 National Institute of Hygiene together with filias in some voivodships up to 1954 were appointed as sanitary epidemiological service especially to control the infectious diseases. In 1954 National Sanitary Inspection was appointed as official institution of central administration in the country to the hygienic and antiepidemiological problems and National Institute of Hygiene stated to work as scientific unit only. National Sanitary Inspection was organised as the net of sanitary-epidemiological stations in voivodeships, districts and as frontiers stations. PMID- 15810499 TI - [Atypical course of chicken-pox complicated by hepatitis and endocarditis]. AB - A case of chicken-pox complicated by hepatitis and endocarditis in 21 years old man was described. Three weeks before admission to the Department of Infectious Diseases the patient stayed at the Neurological Department and was diagnosed as encephalitis. The spots on the skin and a very high level of aminotransferases were noticed in 19th day of hospitalization. The blood cultures were positive for Staphylococcus aureus MSSA. Bacterial endocarditis was diagnosed on the base of echocardiography. The patient was treated with antibiotics six weeks. He recovered completely. PMID- 15810500 TI - [Cardiac manifestations of Lyme borreliosis]. AB - Lyme borreliosis is increasing in Poland in both incidence and recognition with 655 cases reported in 1997 and 3574 in 2003. Approximately 4% of patients will develop cardiac manifestations--the least well documented complication of Lyme disease. Cardiac involvement usually occurs within weeks to months of the infecting tick bite and includes varying degrees of atrioventricular block as the commonest manifestation and tachyarrhythmias, myopericarditis, mild cardiac muscle dysfunction. There has been evidence that long standing dilated cardiomyopathy may be associated with chronic Borrelia burgdorferi infection. Patients with atrioventricular block have good prognosis. Most cases resolve within 1 to 2 weeks. Temporary, but almost never permanent, cardiac pacing may be required for some patients. Cardiac manifestations of Lyme disease are treatable with antibiotics. Lyme carditis should be taken into consideration in patients with acute as well as chronic heart diseases. PMID- 15810501 TI - [Spontaneous bacterial peritonitis in patients with decompensated liver cirrhosis based on bacteriological and biochemical results]. AB - Spontaneous bacterial peritonitis (SBP) is frequent and insidious complication of liver cirrhosis regardless of its aetiology. The aim of the study was to assess the usefulness of biochemical markers in the blood and ascitic fluid, including the concentration of proinflammatory cytokines in the diagnosis of SBP in the patients with decompensated liver cirrhosis. The material and methods: 117 examinations in 88 patients were performed as following: ascitic fluid and blood cultures, throat and anus smears, biochemical examinations in the serum and ascitic fluid including concentration of procalcitonin, TNF-alpha, IL-6 and neopterin. THE RESULTS: 25% of patients have died during the hospitalisation, the positive blood cultures were found in 7 cases (5.9%), whereas the positive ascitic culture in 17 (14.5%) cases. The procalcitonin level were increased in 56.6% of the cases, the remaining levels of cytokines were increased considerably in one case with SBP. Biochemical examinations in the blood, performed on the admission, revealed: hyperbilirubineamia, increased level of ALT, AST, GGTP, alkaline phosphatase, creatinine, WBC and gamma-protein and decreased level of haemoglobin and albumin. CONCLUSIONS: We have obtained Gram positive bacteria in 57% of cases in the ascitic fluid and Gram negative in the 43%. THE RESULTS of the cytokines concentration seam do not have significant importance in the SBP diagnosis. However our study was performed on the small amount of the cases and requires further investigations including the control group. PMID- 15810502 TI - [Microbiological analysis of blood culture obtained from children hospitalized at the Gastroenterology, Oncology and Daily Chemotherapy Wards of the Children's Memorial Health Institute in the years 1999-2002]. AB - The aim of the study was to examine correlations between the use of different kinds of intravascular catheters and the type of microorganisms isolated from blood cultures. Blood samples obtained from gastroenterology, oncology and daily chemotherapy wards were examined. The samples were taken from catheter and peripheral blood in situations where blood infection was suspected. In positive blood samples Gram-positive bacteria, especially methicillin-resistant coagulase negative staphylococci, were dominant. Enterobacteriaceae (Klebsiella sp., E. coli, Enterobacter sp.) were the most often isolated among Gram-negative bacteria. The share of Staphylococcus aureus, Gram-negative bacilli and fungi was greater in the case of samples taken from catheters. The domination of CNS is frequently connected with catheter colonisation or contamination of samples. Intravascular catheters predispose to Staphylococcus aureus, Gram-negative rods and fungal infections. PMID- 15810503 TI - [Coinfection with HIV and HCV--epidemiologic, diagnostic, clinical and therapeutic implications]. AB - Because of shared transmission pathways, coinfection with HIV and HCV is common. Besides evidence is accumulating that HIV infection may be a cofactor for heterosexual and perinatal transmission of HCV infection. The increasing survival of patients on HAART has emphasized the importance of treating liver disease because co-infected patients are now often dying of their liver disease. There is a common opinion that HIV infection causes rapid and more severe course of HCV related liver disease. The impact of HCV infection on HIV disease progression is controversial. The treatment of patients co-infected with HIV and HCV is difficult. Therapeutic options depend on immunological status of patients and severity of liver disease. In addition hepatotoxicity and drug interactions should be considered. PMID- 15810505 TI - [Increase in the incidence of pertussis and quality of whole-cell pertussis component of the DTP vaccine produced in Poland. Part II. Consistency of control and production]. AB - This study aimed to identify and to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine lots produced within 1972 2001 due to the changes having occurred in production and potency testing procedures. Obtained results showed high consistency of the production and control tests confirming relevant quality of country-produced DTP vaccine. PMID- 15810504 TI - [Increase in the incidence of pertussis and quality of whole-cell pertussis component of the DTP vaccine produced in Poland. Part I. Potency of DTP vaccine]. AB - This study aimed to identify and to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine lots produced within 1972 2001 due to the changes having occurred in production and potency testing procedures. The study confirms that higher potency values were obtained for vaccine lots produced since 80-ties, e.g. after changes of: references lots (1975), vaccine strains (1978) and source of animals used in Kendrick tests (1979). Additionally, the comparisons performed revealed a down trend in potency levels within 1992-1999 correlating to the lowering of the number of IOU/dose. PMID- 15810506 TI - [Epidemiology of rotaviral infections among children in Poland]. AB - In Poland there is no obligation of reporting and registration of the rotaviral infections in children. Therefore, data concerning number and incidence of rotaviral infections is unknown. Most of works devoted to rotaviral infections describes clinical course. In many works, Polish authors carry out the analysis of per cent participation of rotaviral infections in global number of children hospitalized due to diarrhea in Poland. The rotaviral part of diarrhea is 20%-80% of all diarrheas in Polish children. Mostly, rotaviral diarrheas take 40% to more than 50% of all diarrheas in children. Rotavirus is also accountable for rising of nosocomial infections. Summarizing, there is conclusion, that is necessary to implement the registration of rotaviral infections in Poland. PMID- 15810507 TI - [Bioactive effectiveness of selected disinfective agents on Gram-negative bacilli isolated from hospital environment]. AB - In our study the susceptibility (MIC) of chosen 21 strains of Gram-negative bacilli isolated in hospitals to disinfectant agents (glucoprotamine, sodium dichloroisocyanurate, potassium persulfate), the effectiveness of these disinfectants against selected bacteria and their effectiveness to biofilm forming bacteria was determined. It was found that glucoprotamine showed the highest activity to Gram-negative bacteria. Obtained MIC values for glucoprotamine (except 1 strain of S. marcescens) were 16-64 times lower that MICs for sodium dichloroisocyanurate and 4-32 times lower that MICs for potassium persulfate. The effectiveness of disinfectants containing potassium persulfate or sodium dichloroisocyanurate was 100% tested by carrier method. Glucoprotamine was ineffective against 2 out of 9 strains (18%): E. cloacae and S. marcescens. It was found that disinfectants were more effective against Gram-negative bacteria in carrier methods than for biofilm forming bacteria. 86% of bacteria growing 5 days on a catheter were resistant to working solution of disinfectant containing glucoprotamine (5200 mg/L) or potassium persulfate (4300 mg/L); 66.6% of tested bacteria were resistant to working solution of sodium dichloroisocyanurate (1795.2 mg/L). In our study the highest effectiveness to biofilm forming bacteria showed disinfectant with sodium dichloroisocyanurate, the lowest--with glucoprotamine. PMID- 15810508 TI - [The role of inflammation in the pathogenesis of atherosclerosis]. AB - Inflammation plays an important role in the initiation and progression of coronary artery disease and the precipitation of acute coronary events. However, the inflammatory triggers are still poorly understood. Histologically, unstable atherosclerotic plaque contains activated macrophages and T lymphocytes, adhesion molecules, chemokines and cytokines, matrix-degrading enzymes and prothrombotic factors. Circulating inflammatory markers such as C-reactive protein (CRP), fibrinogen, and interleukins (especially IL-6) are increased in high-risk groups of patients and predict future risk. PMID- 15810509 TI - [The role of infection in the pathogenesis of atherosclerosis]. AB - Experimental models and human studies have supported a role of infection in the initiation of atherosclerosis. There are many known microorganisms who can play an important role in atherosclerosis, but especially two of them--Chlamydia pneumoniae and Cytomegalovirus are suspected to stimulate the process of atheromatosis. Until antibiotics or vaccines are useful in artery diseases prevention, therapies with proven vascular anti-inflammatory effects (diet, exercise, smoking cessation, aspirin, statins) should be optimized. PMID- 15810510 TI - [Type-2 diabetes mellitus as a cause of disability]. AB - Type 2 diabetes is a metabolic disease associated with a wide range of comorbidities and complications, including retinopathy and loss of vision, nephropathy and end-stage renal disease, peripheral neuropathy, cardiovascular diseases, lower extremity amputations due to diabetic foot. All this long-term medical complications shorten life expectancy of the affected people and cause physical decline, which leads to disability. The importance of the problem results also from the prevalence of the disease. Diabetes is expected to increase worldwide from 135 million to 300 million people between 1995 and 2025. In Poland it is estimated that 2 million inhabitants have diabetes and half of these population is unaware of the condition. The aim of these review of the literature is to show all positive associations between diabetes and its complications leading to limitations and disability. The prevalence of the disability in the population of the people with diabetes is the sum of all diabetic complications and the process of aging. The diabetes epidemic affects the entire age range and the greatest absolute increase is occurring among the elderly. These informations suggest that the number of people at risk for physical limitations and disability, caused by diabetes will increase dramatically. If these process was not stopped, it would erode health status and quality of life of the great proportion of the society. PMID- 15810511 TI - [Effect of smoking during pregnancy on maternal condition and birth outcome- overview of epidemiologic studies]. AB - The association between maternal smoking and retarded fetal grow was first time described in 1957, and now is well know. In addition smoking during pregnancy increases risk of spontaneous abortion, placenta previa, abruptio placenta, preterm premature rupture of membranes, stillbirth, preterm delivery and congenital malformations. The risk for most of these conditions has been found to increase with the number of cigarettes smoked and on the other hand women who stopped smoking during pregnancy are at the lower risk for most of those pathologies. PMID- 15810512 TI - [Incidence of congenital malformations and neonatal mortality in children born at the Regional Hospital in Brzeziny in years 1990-2001]. AB - The aim of the study was to estimate the incidence of congenital malformations and neonatal mortality rate in children born in our hospital from 1990 to 2001, taking into account the effects of low birth weight and premature delivery. We investigated medical documentation of 5496 children born during the analyzed period. Presence of major and minor malformations and survival through the neonatal period were determined for each subject. Among newborns with low birth weight 8.3% were born with congenital malformations, among those born preterm- 12.1%. The major malformations were 3 times more often in children with low birth weight and 4 times more often in children born prematurely. In our cohort the mortality of 0.2% was observed. Congenital defects, followed by the complications of preterm delivery, were the predominant causes of natal death. Deaths due to congenital defects occurred twice as often as the deaths caused by prematurity complications. The congenital malformations mortality in newborns with congenital malformations reached 4.8%. The majority of congenital defects deaths were caused by central nervous system defects, chromosomal aberrations and multiple congenital malformations. PMID- 15810513 TI - [Retrospective overview of medically treated non-fatal injuries in pupils aged 11 15 years in Poland (1994-2002)]. AB - The aim of the study was to analyse: annual occurrence of injuries in pupils aged 11, 13 and 15 years in Poland, common location of injury occurrence and type of activity, as well as the type of body harm. The results of the survey were compared with similar studies in 1994 and 1998 and series of trends were drawn. The survey was carried out in Poland within the Health Behaviour in School-aged Children. A WHO Collaborative Cross-national Study (HBSC). The representative sample (N = 6383) was selected by two-stage, cluster drawing with stratification by place of residence. As the survey instrument the international questionnaire was used, translated and adapted for national purposes. Selected determinants of injuries were examined by crude odds (OR) with 95 per cent confidence interval (CI) calculated from contingency tables with the use of EPI INFO 6.04d software. Results of the study indicated the linear increase of non-fatal injuries among 11, 13, 15 years old pupils in 1994-2002 (from 17.6% to 28.4%) and even more evident increase of repeated injuries in the same time (from 2.1% to 8.6%, respectively). School and home were the most common location of injuries occurrence, while training for sport was the most common type of activity. Boys, living in urban areas were at highest risk of injuries in total, and by selected injuries outcomes. An increase of awareness that injuries are preventable and implementation of well know in the world prevention strategies are needed to stop disadvantageous trends in non-fatal medically treated injuries among school-aged children. PMID- 15810514 TI - [Childhood immunization rate in families of various socioeconomic status in the town of Bytom]. AB - The project of the study was to find out if the childhood immunization rate depends on the social economical status (SES) of the families. The analyzed data were obtained in a population based study performed in 2002 in the town Bytom. Data set included to the health questionnaire provided by families of 13998 children aged from 6-17 years (68% of eligible population. Almost all children (98.5%) attended obligatory vaccination in early childhood. Non-immunization rate (NIR) was associated with self reported SES ("poor": 2.7% NIR, "good": 0.7% NIR; p < 0.0001). Other correlates included family income (p < 0.0001), maternal education (p < 0.0001), and family size (p < 0.0001). The multivariate logistic analysis confirms the effect of SES variables on the childhood immunization rate. PMID- 15810515 TI - [Epidemiologic trends of the intake of psychoactive agents by basic service soldiers]. AB - The aim of the investigations described in this study is to present epidemiologic tendencies of drug addiction in the army and to define its causes and conditionings. The investigations were carried out in the years 1996-2003 among 3561 basic service soldiers and 134 members of the cadre, with the use of the same questionnaire, which allowed to compare the results from respective. The carried out analysis of epidemiologic trends in basic service soldiers taking psychoactive agents proved the downward tendency of drug addiction in the army. This decrease has been occurring since 2000. The proportion of soldiers taking psychoactive agents decreased twice in 2003 in comparison with 1996 and four times in comparison with 2000. In the last three years there occurred a decrease of percentage of basic service soldiers taking psychoactive agents and there decidedly increased the number of prophylactic actions. PMID- 15810516 TI - [Features of the Arab health service as exemplified in Lebanon]. AB - The Arab health service profile on Lebanon example, one of the Middle East countries is presented in this article. Nowadays the world becomes a global village, a free flow of people appears, who travel professionally as well as touristically. In situations of health and life hazards, in cases of diseases and injuries caused by traumas arisen in foreign countries it seems to be important to get to know problems concerning a health service condition in the region of the world being a place of work or rest. For a long time region of the Middle East have been situated in the center of interests of the world public opinion. On the one hand it is an attractive place because of tourist reasons, on the other hand take place an escalation of military conflicts in individual Arab and Muslim countries. Because of this situation permanent information on the subject of health care status in the countries of temporary stay where occurs unstable political and military situation it seems to be a necessity. PMID- 15810517 TI - [General principles of database storage operations with emphasis on OLAP reports]. AB - In article general principles and features of data warehouse were presented in particular of OLAP reports. The data warehouse was built using Oracle tools. The repository was filled with death records from Central Office of Statistics. Various features adequate for epidemiological analyses have been discussed and illustrated like pivoting and rotating dimension, drilling on hierarchical data, reduction of dimensions. The possibility of specific for epidemiology indicators creation was shown. The need of implementation of data warehouses and OLAP reports in Polish healthcare was discussed. In comparison with traditional manner of analysis and presentation epidemiological facts OLAP reports give new perspectives. PMID- 15810518 TI - [Results of dynamic cytogenetic study of children and teenagers living in areas, radioactive by contaminated after the Chernobyl accident]. AB - Results of multiyear cytogenetic study of children and teenagers living in areas, radioactive by contaminated after Chernobyl accident, were adduced. Mean density of 137Cs contamination in two compared living areas were 111 and 200 kBq/m2 and mean external doses accumulated for 1986-2001 were 6.7 and 11.4 mGy correspondingly. Averaged thyroid doses receives by subjects of all age groups in the second area were approximately 1.5 times higher than in the first area; in the youngest group (0-1 year) the doses were 114.3 and 174.3 mGy. During 17 years cytogenetic investigation approximately from 30% to 60% of examined persons were observed the increased level of chromosome aberrations in lymphocytes of peripheral blood. Average frequency of unstable aberrations (acentrics, dicentrics and centric rings) constituted about 0.4 per 100 cells (0.22 per 100 cells in controls) during all period of observation. Level of marker aberrations (dicentrics and centric rings) was increased almost all times of study and varied within 0.04-0.19 per 100 cell (0.03 in control group). The parallel investigation of frequency of stable aberrations by FISH method showed up their level about 3 times exceeding observed dicentrics level. Comparably higher indexes of cytogenetic disturbances were revealed in group exposed in utero during period of accident. PMID- 15810519 TI - [The use of computer system in the introduction of countermeasures in agriculture after the Chernobyl accident]. AB - In the development of strategies for countermeasures aimed at the rehabilitation of contaminated areas, essential is to identify criteria for assessing their effectiveness and factors influencing the priority of the criteria chosen. A method is suggested for assessing effectiveness of countermeasures in agriculture based on a multicriterium analysis of radiologic, economic, regulatory and socio psychological indicators characterizing the use of countermeasures. Rating of countermeasures strategies is presented depending on financing of works on the rehabilitation of the Chernobyl affected areas. PMID- 15810520 TI - [The role of regulatory networks of the cell response to damages in radiation effects]. AB - In view of modern knowledge and concepts about components, function and mechanisms of response of cell molecular structures to damaging effects, response which is generating specialized modules of reactions, it is shown that main components of the mechanism of maintenance of genome constancy at ionizing radiation exposure are checkpoints of cell cycle, DNA repair and apoptosis. They operate under the control of a genetic system at participation of Tp53 gene, corresponding protein and of regulatory networks formed by cascades of mitogen activated protein kinases (MAPK). At ionizing radiation exposure the MAPK special modules participate in formation of radiation effect: ERK 1/2 (extracellular signal-regulated kinase 1 and 2), JNK/SAPK (c-Jun N-terminal kinase/stress activated protein kinase) and p38 MAPK. Executing physiological functions of maintenance of normal life activity of cells, they do not lose this capacity after exposure to ionizing radiation, participating in formation of radiation effect in a wide range of doses, and are inactivated only by exposure to very high doses. It is concluded that in light of the modern data the main problem is not a problem of mechanisms of biological effect of ionizing radiation but a problem of biological mechanisms of radiation exposure. PMID- 15810521 TI - [The role of initial state of adrenal cortex in external irradiation effects upon its structure, function and androgen-receptor interactions]. AB - We studied morphological-biochemical time-course reactions of adrenal cortex and hormonal function of testicles after combined or separate actions of external radiation exposure (1.0 Gy at 7.8 x 10(-4) Gy/sec) and prolonged (12-, 32 days) subcutaneous corticosterone injections (10 mg/kg/animal) in 2-3-month old male rats. Both variants with exogenous hormone depressed endogenous glucocorticoid production and evolved acute structure-functional adrenal gland cortex deficiency. Radiocensitivity of the gland at the experimental conditions was increased. Simultaneously the aggravation of adrenal cortex functional activity accompanied with hypogonadism and depressed reproductive potential of male rats, accessed by means of androgen status and androgenspecific (transporting and receptional) proteins characteristics as sensitive molecular markers of an animal possible fecundity. PMID- 15810522 TI - [Proteolytic processes in lung tissue of rat posterity after whole-body chronic irradiation with low doses]. AB - Intensification of proteolytic processes accompanied by inhibited antiprotease activity in lung tissue of first generation posterity of rats irradiated with a total dose of 0.25 Gy (0.01 Gy per day) before gestation was shown. The obtained data satisfied to imbalance in functioning of regulation systems, tension of adaptation mechanisms with possible rapider reduction of adaptation resistance of broncho-lung system of irradiated parents' posterity. Vector changes in correlation of indexes in the proteinase-antiproteinase system were analogous in all experimental series (irradiated male x irradiated female, irradiated male x intact female, intact male x irradiated female) with maximal expression in the group obtained from parents both irradiated before gestation. It could be possible to propose a presence of non-stable genome in posterity of irradiated parents, that satisfied to role of genetic aspects in development of the late functional disorders. PMID- 15810523 TI - [Genotoxic and radioprotective properties of 2,5-diphenyloxazole and its derivatives on mice DNA structure in vivo]. AB - The effects of 2,5-diphenyloxazole ad its derivatives suggested as nontraditional radioprotectors on mice spleen DNA structure were studied. The effects of these compounds were studied on intraperitoneal injection with and without subsequent X ray exposure of mice to a dose of 12 cGy. The formation of double-strand breaks and DNA conformation change (by adsorption on NC filters) were recorded. A genotoxic effect of 2,5-diphenyloxazole and its derivatives, a nonlinearity of their dose-response relationships and different effects depending on the substance concentration were found. Some of the compounds exhibited radioprotective properties in certain concentration. PMID- 15810524 TI - [Modification of cellular radiosensitivity by NO-synthase inhibitors]. AB - We recently reported that the treatment of V-79 and HeLa cells with nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) significantly reduced the level of the radiation-induced unstable chromosome aberrations. The stereoisomer D-NAME had no effect. We suggest that the radioprotective effect of L-NAME resulted from the action on the generation reactive radicals due to the inhibition of the NOS-activity. We tested this suggestion on the NO-resistant (ECV-304) and NO-sensitive (HeLa) cells, which were treated with L-NAME or aminoguanidine or D-NAME or cysteamine before gamma-irradiation. There are no significantly differences in radiosensitivity between these cells estimated after exposure by gamma-rays with different doses. However, the radioprotective effect of the NOS-inhibitors manifested only for HeLa. D-NAME had no radioprotective effect neither HeLa nor ECV-304. In contrast NOS-inhibitors, cysteamine treatment EVC-304 reduced the radiation-induced level chromosome aberrations almost twofold. The different mechanisms of the modification of cellular radiosensitivity are discussed. PMID- 15810526 TI - [Estimation of the influence of physical and biological factors on the development of the hematopoietic type of radiation sickness in dogs and two types of monkeys]. AB - In this investigation, the analysis of radiobiological experiments on 532 dogs and two types of monkeys (101 animals), irradiated totally in the 1.0 to 6.0 Gy dose range at different irradiation facilities, has been carried out. LD50 values at X-ray and gamma-neutron exposure were close to each other (2.35 and 2.83 Gy, respectively) while at gamma-radiation exposure LD(50/45) increased to 3.09 Gy. Comparison of LD(50/45) values for different kinds of animals allowed us to draw a conclusion of approximately equal radiosensitivities of dogs and Macaca fascicularis monkeys (LD(50/30-45) - 3.09 Gy and 3.17 Gy, respectively); Macaca rhesus monkeys revealed higher radioresistance (LD(50/30-45) - 5.03Gy). Analysis of the influence of several biological factors has not displayed any significant differences in the values of LD(50/45) and average lifespan of male and female dogs. Higher radiosensitivity of dogs with body weight less than 12 kg and lower radiosensitivity of dogs in summer time compared to other seasons have been shown. Dogs at the age of 2 to 3 years appeared to be more radioresistant than animals of the other age. PMID- 15810525 TI - [Radioprotective and antistressful properties of nitric oxide production modulators]. AB - The radioprotective and antistressful activities of L-arginine and the "Pronumol" preparation, in which L-arginine is contained in the complex of proteins with nucleic acids, were studied. In mice repeated peroral intake of L-arginine and "Pronumol" partially prevented radiation-induced and stress-induced lipid peroxidation and DNA degradation in thymus, increased hemopoietic stem cell survival, and prevented an increase in chromosome aberration frequency in bone marrow cells of irradiated mice. When repeatedly administered per os before irradiation, "Pronumol" increased survival of intestinal stem cells in irradiated mice and prevented thymus cell devastation induced by radiation and stress. PMID- 15810527 TI - [Quantitative regularities and peculiarities of the development of the cerebral form of radiation sickness at fractionated irradiation]. AB - Several peculiarities in manifestations of cerebral form of radiation sickness have been revealed at a fractionated double irradiation with equal and unequal doses per fraction and different intervals between the fractions. A reliable increase in average lifespan of rats irradiated with (100 + 100 Gy) equal doses at 10 and 60 min intervals between two fractions compared to the single radiation exposure to 200 Gy has been obtained. Lifespan of rats irradiated with a total dose greater than 200 Gy in most cases of double exposures with 10 min interval was reliably less than that for animals after a single exposure. The influence of the first dose on the reduction of animal average lifespan increased with fraction dose increasing from 150 to 300 Gy and was most pronounced at the total exposure dose of 400 Gy. Reaction of rats on the repeated irradiation was significantly weakened in comparison with the reaction on the first exposure. At a study of capacitation the interval of 30 min appeared to be more favorable compared to 10 min interval. Importance of a dose value in the first fraction has been demonstrated: the higher this value the worse the capacity of the rats 3 hours after the repeated exposure. PMID- 15810528 TI - [Long-term morphofunctional changes in the skin of Chernobyl clean-up workers]. AB - Musculocutaneous samples taken from 19 Chernobyl clean-up workers were morphofunctionally examined to trace the changes, which had happened in them under the lasting exposure to low doses of ionizing radiation. The analysis showed that the greatest changes had taken place in the epidermis in the form of thickening of corneal and cellular layers and inflammatory infiltration of lymphocytes provided with the productive panvasculitis in the majority of arterioles. It is supposed that after the long low-doce irradiation two forms of reactions developed in the skin: a defense reaction in the form of proliferative hyperkeratosis and an immunopathological reaction. The latter is a consequence of activation in the cellular layer of epidermis combined with the appearance of effector component of immune response, which stimulates the interaction of epidermal T-lymphocytes with the endothelial cells of derma vessels and systemic involving of blood vessels of microcircular blood stream. PMID- 15810529 TI - [Evaluation of cytophysiological state of testis in forest voles living in the radioactively contaminated areas]. AB - We made a cytophysiological analys of testis in Clethrionomus rutilus and Apodemus sylvaticus inhabiting a contaminated area of the East Ural Radioactive Trace (EURT). The study showed that in the norm these species differed in the total number of cells of spermatogenous epithelium, due to interspecific difference. In the sampling plot a concentration of 90Sr in Clethrionomus rutilus was 2 times higher than that in Apodemus sylvaticus. Maximum destructive changes in endocrine and exocrine (seminiferous tubules) section of testis and increased proliferation activity of spermatogenous epithelium were observed in Clethrionomus rutilus.Increased proliferation activity was found as a compensatory-protective reaction which promote the maintenance of the germ cell number. Such changes were not observed in Apodemus sylvaticus. PMID- 15810530 TI - [Current radiation status of the ecosystem of the Lake Bolshoi Igish situated along the axial part of the East-Urals radioactive trace]. AB - Data on the current radioecological status of lake Bolshoi Igish in the Middle Ural are presented in the paper. The lake is situated on the central axis of the East-Urals radioactive trace which was formed as a result of the 1957 nuclear accident at the Mayak Production Association. The distribution of 90Sr and 137Cs in the major components of the water basin is shown. Seasonal dynamics of the specific activity of radionuclides in water and in bottom deposits has been investigated. The current community of the higher water plant species has been studied. The paper discusses the concentration of 90Sr and 137Cs in the macrophyts of lake Bolshoi Igish depending on their species. PMID- 15810531 TI - [Prognosis of accumulation of 137Cs and 90Sr in the herbage of the main types of the Belarus Polessje meadows using agrochemical soil properties]. AB - On the basis of long-term stationary experience it was established that the minimum accumulation quantities for 137Cs and 90Sr in the herbage of the dry, lowland and flood-plain types of the Belarus Polessje meadows contaminated with Chemobyl radionuclides are determined when the optimum of basic agrochemical soil properties is achieved with application of the scientifically reasonable protective measures. For remote prognosis of radionuclide contents in natural and cultural meadow herbage the use of transfer factors (TFa, (Bq/kg)(kBq/m2)) based on the complex agrochemical parameters--agrochemical cultivation soil index (Icd) and basic saturation degree (V, %), which take into account some soil characteristics simultaneously, is a streamlined approach. This paper provides the equations of linear and multiple regressions, which can be used to calculate the transfer factors for 137Cs and 90Sr uptake and the herbage contamination degree for the main types of meadows of the region, that will allow reducing the volume of forage production (hay, green bulk), which is not adequate to established permissible levels: "Republican allowable levels of the contents of 137Cs and 90Sr in agricultural raw material and forages". PMID- 15810532 TI - [Analysis of the effects of low-intensity intermittent and continuous electromagnetic fields on behavioral acts in the mice]. AB - The fact of modulating influence of low-intensity electromagnetic field on the behavioral acts in mice was established. The dependence of mice behavior on the modulating frequency, electromagnetic field parameters (intermittent or continuous) and on the time of adaplive reaction was found. PMID- 15810534 TI - [The 27th General Assembly of the International Union of Radioecology]. PMID- 15810535 TI - [International Congress of Radioecologists]. PMID- 15810536 TI - [International Scientific Workshop "Radioecology of Chernobyl Zone"]. PMID- 15810538 TI - [International Conference "Mobile Communication and Health: Medical, Biological and Social Problems"]. PMID- 15810539 TI - [Medicus homo est. Monitoring medical error. Collegium Medicum Uniwersytetu Jagielloskiego]. PMID- 15810540 TI - Monitoring medical error. PMID- 15810541 TI - [Medical error and patients rights]. PMID- 15810542 TI - [Quality monitoring in health care]. PMID- 15810543 TI - [Medical error. Experience of the professional responsibility advocate]. PMID- 15810544 TI - [Medical error and experience of the patients rights advocate]. PMID- 15810545 TI - [Dog-days, cold periods, grape-harvests (France, 15-19th centuries)]. AB - Dog-days, cold periods, grape-harvests (France, 15-19th centuries). The climate history can be based on several kinds of data. In the present paper, French records of grape-harvest dates in Burgundy from 1370 to 1890 were used for evaluate the climates at these various period of time. These results reveal that temperatures as high as those reached in the 1990s have occurred several times in Burgundy since 1370. Correlations between temperatures and historic data are presented. PMID- 15810546 TI - A chimera encoding the fusion of an acetylcholine-binding protein to an ion channel is stabilized in a state close to the desensitized form of ligand-gated ion channels. AB - To understand the mechanism of allosteric coupling between the ligand-binding domain and the ion channel of the Cys-loop ligand-gated ion channels (LGICs), we fused the soluble acetylcholine-binding protein (AChBP), which lacks an ion channel, to either the cationic serotonin type-3A ion channel (5HT(3A)) or the anionic glycine ion channel. Both linear chimeras expressed in HEK-293 cells display high affinity for the nicotinic agonist epibatidine (K(D) = 0.2-0.5 nM), but are not targeted to the cell surface. Only after substituting a ring of three loops located at the putative membrane side of the AChBP three-dimensional structure by the homologous residues of 5HT(3A), the resulting chimera AChBP(ring)/5HT(3A) (i) still displayed on intact cells an apparent high affinity for epibatidine, yet with a fourfold decrease (K(D) = 2.1 nM), (ii) displayed a high proportion of low affinity sites (11 +/- 7 microM) for the resting state stabilizing competitive antagonist alpha-bungarotoxin and (iii) was successfully targeted to the cell surface, as seen by immunofluorescence labelling. The AChBP(ring)/5HT(3A) chimera forms a pentameric structure, as revealed by sucrose gradient sedimentation. However, no whole-cell patch-clamp currents were detectable. Interestingly, binding assays with membrane fragments prepared from cells expressing AChBP(ring)/5HT(3A) showed a decrease in the apparent affinity for the agonists nicotine and epibatidine (5-fold), concomitant with an increase in the proportion of high-affinity sites (48 +/- 1 nM) for alpha-bungarotoxin. These results indicate that fusion of AChBP to an ion channel forms a pentameric receptor exposed to the cell surface and able to convert between discrete allosteric states, but stabilized in a high affinity state for epibatidine that likely corresponds to a desensitized form of LGICs. These artificial chimeras might offer a useful system to investigate signal transduction in LGICs. PMID- 15810547 TI - Feeding preferences of the Apollo butterfly (Parnassius apollo ssp. frankenbergeri) larvae inhabiting the Pieniny Mts (southern Poland). AB - Parnassius apollo (Lepidoptera, Papilionidae) is considered to be typical stenophagous species. Its European forms fall generally into 'telephiophagous' or 'albophagous' trophic groups. According to some authors, 'telephiophagous' P. apollo ssp. frankenbergeri SLABY, inhabiting the Pieniny Mts, has a rather broad spectrum of food-plants. We aimed at defining its feeding preferences for successful breeding in a semi-natural colony on more than one commercially available Sedum species. Larval development (L5) and performance were assessed in three experimental groups fed on different plant species selected in a preliminary test. Apollo larvae appeared to be quite specific in the plant choice and developed poorly in the absence of Sedum telephium. Possible reasons of this are discussed. PMID- 15810548 TI - Destructive and non-destructive microanalysis of biocarbonates applied to anomalous otoliths of archaeological and modern sciaenids (Teleostei) from Peru and Chile. AB - Anomalous otoliths were discovered among modern and archaeological (8th millennium BP) sciaenids. The two species concerned, Cilus gilberti and Sciaena deliciosa, are common on the Peruvian-Chilean coast and do not seem to be affected by this morphological anomaly that maintained in their populations for thousands of years. The carbonates of the anomalous forms, determined by X-ray diffraction, are different from that of the normal otoliths, i.e. calcite and vaterite instead of aragonite. A method of non-destructive analysis by cathodoluminescence is tested and assumptions on the origin of the anomaly and its possible implications on environmental studies are advanced. PMID- 15810549 TI - [In vivo impedance of the aerial organs of some mono- and dicotyledonous plants]. AB - In vivo impedance of the aerial organs of some mono- and dicotyledonous plants. We measure in this paper the electrical capacitance and resistance of aerial organs of some plants, grown in the laboratory (sunflower, pumpkin, and maize, of different ages) or bought at the market (sorrel), in a range of 34 frequencies, from 40 Hz to 100 kHz. The electrospectrometric curves of the leaves aged 14 days of the dicotyledoneous studied are close to each other. PMID- 15810550 TI - Influence of temperature and nutritional requirements for mycelial growth of Eutypa lata, a vineyard pathogenic fungus. AB - Eutypa dieback (dying arm disease, eutypiosis) is a very devastating disease in many grape-producing areas around the world. This vascular disease is induced by the ascomycete Eutypa lata Pers. Fr. Tul & C. Tul. invading the trunk by pruning wounds. The environmental factors and the nutritional requirements regulating fungus development are yet poorly known. This work shows that the isolated strain of E. lata was able to grow in a large temperature range (2-30 degrees C). However, a higher temperature (35 degrees C) presented inhibitory effects on mycelial growth. E. lata was able to use various osidic molecules (C5, C6, C12, C18, C24, and starch); showing thus a large adaptation to the carbon source supplied. As nitrogen source, it used salts and numerous natural amino acids. A significant result was obtained with cysteine presenting obvious antifungal properties. This effect can further be used with the aim of setting up a curative treatment of the disease. PMID- 15810551 TI - Production of NA by endothelial NO-synthase: an in vitro versus in vivo study. AB - Endothelial-derived relaxing factor (EDRF) is secreted by different endothelia in vivo. It is synthesised by endothelial NO-synthase (eNOS). Despite numerous works, its identity is not fully understood. Here the production of NA, a nitroso arginine, which was shown to be synthesised by brain NO-synthase (bNOS), was studied in eNOS preparations. NA was quantified by reductive differential pulse voltammetry (RDPV) during its irreversible electrochemical transformation to N hydroxy-arginine (NHA). Using microelectrodes, NA and nitrite were simultaneously measured in pure recombinant eNOS giving similar enzyme activity. NA was detected at the surface of human endothelial cells (HUVEC) and disappeared when D-arginine was introduced in the culture medium. NA production by endothelium tissue was studied in rat corpus cavernosum using voltammetric microelectrodes. NA concentration at the endothelium surface was linked to vasodilatation measured by laser Doppler induced by acetylcholine injection. LNMA ic injection induced NA disappearance. These preliminary new experiments suggested that NA could be the endogenous nitroso-compound presented early as EDRF. PMID- 15810552 TI - [Comparative evaluation of the toxicity induced by mussels (Mytilus galloprovincialis) from two sites of the Moroccan Atlantic coast in mice]. AB - Atlantic coast in mice. Preliminary studies showed that seawater contains heavy metals from domestic, agricultural and industrial wastes. Marine bivalves concentrate these pollutants by filtration and serve as vectors in human exposure. The objective of this study was to determine the concentration of heavy metals; cadmium (Cd); chromium (Cr), and lead (Pb) in mussels (Mytilus galloprovincialis) collected in two coastal sites; Jorf Lasfar (JL) (neighbouring a phosphate processing platform) and Oualidia (OL) (a vegetable growing area) located at 120 and 190 km south of Casablanca, respectively. Another objective was to test and compare the toxicity of these mussels on mice. The results indicated the presence of heavy metals (Cd, Cr, and Pb) in mussels at different concentrations, depending on the collection period. Higher concentrations were obtained at JL than at OL: for example, Cd concentrations were 80 +/- 15 to 199 +/- 28 versus 23 +/- 5 microg/g mussel dry weight, respectively. Cramming with mussel powder did not increase Cd, Cr, or Pb concentration in either liver or kidneys of treated mice. The relative kidney weights were reduced. Increased glucose urea was observed in animals' urine. Treatment with mussels from OL induced significant reduction (20%) in mice body weight, together with an increase in creatinuria. These results indicate that mussels collected from OL are more harmful than those obtained from JL are. All these mussels should not be recommended for human consumption. PMID- 15810553 TI - Unexpected response of the seagrass Posidonia oceanica to a warm-water episode in the north western Mediterranean Sea. AB - The response of Posidonia oceanica (Linnaeus) Delile to the warm-water episode of summer 1999 was studied by means of the technique of lepidochronology. Study sites include three sites affected by the mass mortality event of benthic invertebrates and one not affected. The results showed a significant decline in some parameters (number of leaves and/or rhizome growth) for the three sites affected by the mass mortality event for the year following the warm-water episode (1999-2000). A similar decline was not observed for the unaffected site. The fact that high temperatures could have a negative impact on deep Posidonia oceanica near its cold limit of distribution is an unexpected result. PMID- 15810554 TI - [Defensive reactions of gastropod molluscs. Humoral reactions]. AB - Humoral protective reactions and their mechanisms in gastropod mollusks are considered based on the results of various investigations. It is important to note that lectines in molluscs, as well as in other invertebrates, are functional analogues of immunoglobulins in vertebrates. PMID- 15810555 TI - [Parasite fauna in sticklebacks (Gasterosteidae) from bodies of water of the Kola region]. AB - The results of a parasitological study of the two species of sticklebacks (Pungitius pungitius and Gasterosteus aculeatus) inhabiting the Kola region are presented. The 42 and 14 species of parasite were found in the nine-spined and three-spined stickleback, respectively. The paucity of the parasite fauna in the three-spined stickleback is observed. Distribution of parasites in Kola waterbodies and ecological peculiarities in system "parasite-host" are presented. PMID- 15810556 TI - [Parasite fauna in salmonidean fishes from the Paanaiarvi-Olanga lake-river system]. AB - The data on parasite fauna in the brown trout Salmo trutta, white fish Coregonus lavaretus, and grayling Thymall thymallus from the Paanajarvi-Olanga lake-river system (White Sea basin) are presented. As like-river systems in this area remain in natural conditions, without any anthropogenic pressure, the parasite fauna in salmonoideans from these water bodies had not been changed and has an archetypical state. The parasite fauna core is established in fishes aged 3+. Parasite species forming the core of the parasite fauna infected the brown trout, grayling and whitefish in different degree. PMID- 15810557 TI - [A preliminary study of parasite diversity in the anadromous Kamchatkan mikizha Parasalmo mykiss mykiss (Osteichthyes: Salmoniformes: Salmonidae)]. AB - The study of parasites associated with the anadromous mikizha from several rivers of the Western Kamchatka has revealed 44 parasite species. The majority of parasites (59%) belong to marine and estuarine-marine species. Lecithophyllum bothryophorum, Echinorhynchus leidyi and plerocercoids of the family Litobothriidae were found for the first time in anadromous fish entering into Asian continental watersheds. It has been revealed that only a few specimens of the anadromous mikizha feed in fresh waters. Most of feeding fish consume a minor amount of food, and this feeding is occasional. Young forms of intestinal parasites of the freshwater and estuarine-freshwater groups (Neoechinorhynchus spp., Crepidostomum spp., Cucullanus truttae, Eubothrium salvelinf) come into anadromous fishes mainly before their entering in fresh waters. PMID- 15810558 TI - [Distribution of trematodes of the family Prosthogonimidae in river and lake ecological systems in the south of the Western Siberia]. AB - The results of long-term investigations (1994-2003) of an infection rate of trematodes of the family Prosthogonimidae in the first intermediate hosts (snails of the family Bithyniidae) and in the final hosts (birds) from a basin of the Chany Lake (Western Siberia) are discussed. A total of 1824 specimens of Bithynia tentaculata (L., 1758) and 7166 specimens of Opisthorchophorus troscheli (Paasch, 1842) have been collected from eight rivers and two lakes in the south of the Western Siberia. Data of incomplete helminthological dissection of 225 young water-fowl of 18 species were analyzed. Birds were obtained in July, August and September, 1996-2003 from the Chany lake basin. Maritas of the trematode family Prosthogonimidae were revealed in bursa Fabricii (26.12%) of seven bird species of two orders (Anseriformes and Gruiformes): Grey Lag-Goose Anser anser (L.) (Infection rate-28.6% +/- 12.1), Mallard Anas platyrhynchos L. (27.5% +/- 6.2), Shoveler--A. clypeata L. (22.2% +/- 13.9), Pochard Aythya ferina (L.) (51.7% +/- 9.3), Coot Fulica atra L. (28.6% +/- 5.6), Tufted Duck A. fuligula (L.) (1 case), Wigeon A. penelope L. (1 case). Average intensity of invasion is 5.8 marita specimens per infected bird. The maximal number of parasites (37 maritas), and also the highest average intensity of invasion are recorded for the Coot. The 342 maritas of the family Prosthogonimidae belong to three species: Prosthogonimus ovatus (Rud., 1803) (49.7%), P. cuneatus (Rud., 1809) (32.8%) and Schistogonimus rarus (Braun, 1901) (14.6%); these trematode species have been recorded in 35, 12 and 8 bird species, respectively. Grey Lag-Goose, Shoveler, Pochard and Tufted Duck are new host records for P. cuneatus in for Western Siberia. Parthenitae of trematodes (Prosthogonimidae) were revealed from eight populations of bithyniid snails. The infection rate of the trematodes (parthenitae and cercariae) in population of bithyniid snails from lake ecological system varied 6.55-54.4%, and in river ecological systems in was 0.6-4.2 %. The infection rate of trematodes (parthenitae and cercariae) of the first intermediate hosts was 2.64% that is significantly lower than that of the final hosts from the Chany Lake basin. Both O. troscheli and B. tentaculata were noted as the first intermediate hosts of prosthogonimid trematodes of three species: P. ovatus, P. cuneatus and S. rarus. All three species of trematode were found in three water reservoirs: Ob River (B. tentaculata), Kargat River (O. troscheli) and Chany Lake (O. troscheli). Thus it is found out that 39 bird species can be the final hosts of prosthogonimid trematodes. About a quarter of nestlings is infected with these trematodes. The infection rate of parthenitae in snails from the lake ecosystems is significantly higher than in the river ecosystems. PMID- 15810559 TI - [Seasonal dynamics of diurnal production and rhythmic of diurnal production of the cercaria Diplostomum volvens Nordmann, 1832 in molluscs of the genus Lymnaea from lakes of the Karasuk water system]. AB - Infection rate of the cercariae Diplostomum volvens in molluscs of the genus Lymnaea in natural conditions (Western Siberia) was examined in 1990-1994. Experimental studies have shown that light and water temperatire are main factors regulating rhythmic of diurnal production of cercariae by molluscs. PMID- 15810560 TI - [Population of the mosquitoes Culex pipiens molestus (Diptera: Culicidae) in dwellings of Lensk city]. AB - Culex pipiens molestus has been distributed in Siberia and the Russian Far East since 80s of XX century. In early 90s, C. p. molestus reached South Yakutia (Nerungri city), and nowadays it occurs in South-West Yakutia (Lensk city). The number of breeding places with larvae and pupae of mosquitoes was not numerous in Lensk; among 56 examined houses, the places with larvae and pupae mosquitoes were registered in seven houses, while the adult mosquitoes occurred in the majority of the houses. The occurrence of larvae varied from 12.5 to 33.3%, occurrence of adults-from 12.5 to 100%. The coefficient of variation of larvae in the samples was equal to 0.92, and adults had the following coefficients: 0.84 in the houses without basement; 0.92 in the houses with dry basements; 1.05 in the houses with pools in basements. This data are referred to the casual type, it means that the distribution of mosquitoes in space is dispersed. Imago spread actively in stone house of Lensk that provides mosquitoes with new places of breeding, and supports the stability of population in new areas. PMID- 15810561 TI - [A description of a new species Gyrodactylus moldovicus sp. n. (Monogenea: Gyrodactylidae) from the European mudminnow Umbra krameri Walbaum, 1792 from the lower Dnester basin]. AB - Gyrodactylus moldovicus sp. n. found on gills, body and in nasal cavities of the European mudminnow (Umbra krameri) differs from G. slovacicus Ergens, 1963 also living on the this host by bigger size of the body, anchors and marginal hooks; from G. cylindriformes Mueller et Van Cleave, 1932 living on the American mudminnow Umbra limi--by bigger size of the body; from G. limi Wood et Mizelle, 1957 also from U. limi--by the form of ventral and dorsal bars and form of marginal hooks. It differs from other freshwater gyrodactylids by special type of marginal hooks which have a hook-like end of the blade. Gyrodactylus moldovicus, G. slovacicus and G. limi have marginal hooks of quite different morphological types. By the morphology of anchors, ventral and sometimes dorsal bars and also morphology of cirrus, G. moldovicus is most related to three species from Cyprininae: G. stankovichi Ergens, 1970, G. longoacuminatus Zitnan, 1964 f. typica and G. shulmani Ling, 1962. PMID- 15810562 TI - Stress perceptions in community clinic: a pilot survey of patients and physicians. AB - This pilot survey of 103 patients and 17 physicians in an urban family medicine clinic gathered information pertinent to the design of a feasible stress intervention for patients bothered by stress, but who did not have a psychiatric diagnosis. Among patients, 45% reported being excessively bothered by stress in the preceding month, with the chief stressors being job (70% reporting), financial worries (58%) and family concerns (50%). Patients reported a variety of problems perceived to be related to stress, such as headaches, insomnia, eating control, and gastrointestinal symptoms. Although about 80% reported using positive coping methods (e.g., talking, exercising, and relaxing), 42% reported using alcohol, and 10% used non-prescribed drugs to cope with stress. Only 37% of patients had sought help for stress from their physician. The wide variety of responses from the physicians reflected a lack of standardized approaches, inadequate training, and a reluctance to engage patients about their stress problems. About 42% of the physicians reported routinely asking patients about stress, and 77% felt that dealing with patient stress was a significant burden on their practice of medicine. Overall, the findings indicate that opportunities are being missed for helping patients to deal with stress constructively, and that a standardized stress self-management program might be one solution. PMID- 15810563 TI - An evaluation of independent consumer assistance centers on problem resolution and user satisfaction: the consumer perspective. AB - Individuals who wish to receive independent assistance to resolve access to care health problems have limited options. The Health Consumer Alliance (HCA) is an independent, coordinated effort of nine legal services organizations that provide free assistance to low-income health consumers in 10 California counties. The need for the HCA stems from the vast number of health consumers with unanswered questions and unresolved problems relating to access to care issues, among both insured and uninsured populations. However, little is known about the effectiveness of independent consumer assistance centers. This paper examines the effectiveness of a network of independent consumer assistance programs in resolving consumer problems and consumers' level of satisfaction with services received. As the project evaluators, we conducted telephone surveys with 1,291 users of the HCA to assess if this independent program resolved consumer problems, and to measure the level of satisfaction among HCA users. Specifically, we asked questions about the HCA's influence on problem resolution, consumer satisfaction, health insurance status and use of preventive care services. From 1997 to 2001, more than 46,000 consumers contacted the seven health consumer centers (HCCs). According to our sample of respondents, results show that the HCCs are an important resource for low-income Californians trying to access health care. After contacting the HCCs, 62 percent of the participants report that their problems were resolved. In addition, 87 percent of the participants said the HCCs were helpful and 95 percent said they would be likely to contact the HCC again if necessary. PMID- 15810564 TI - Physical activity, cardiorespiratory fitness, and their relationship to cardiovascular risk factors in African Americans and non-African Americans with above-optimal blood pressure. AB - This report describes cross-sectional associations among physical activity, cardiorespiratory fitness, dietary habits, and cardiovascular disease (CVD) risk factors in a large sample (n = 810) of African Americans (n = 279) and non African Americans (n = 531) with above-optimal blood pressure. Participants in PREMIER, a clinical trial for blood pressure control through lifestyle approaches, underwent baseline assessments to determine physical activity level, cardiorespiratory fitness category, dietary intake, and CVD risk factors. Mean levels of body mass index (BMI), total cholesterol, LDL cholesterol, HDL cholesterol, daily percent calories from fat and saturated fat, daily servings of fruits and vegetables, and daily fiber intake were examined across three physical activity levels and two fitness categories. Hypertension status was also assessed. Data were stratified by sex and ethnicity. For all participants, those in the low fitness category had higher BMI levels. Total cholesterol was lower in African American women in the high fitness category. Mean values of more than five daily servings of fruits and vegetables were reported by non-African American women and African American men in the high activity category. Higher intake of dietary fiber was found for non-African American women at the high activity level, with a similar trend observed for African American women. Future work examining these associations prospectively should include sufficient minority representation to enhance generalizability to all population groups and determine the beneficial effects from increased physical activity and improved cardiorespiratory fitness. PMID- 15810565 TI - Psychosocial problems among patients in neighborhood health centers: perspectives from health care providers. AB - Little is known about how health care professionals perceive and understand the psychosocial problems of individuals receiving services in neighborhood health centers (NHCs). We conducted interviews with health care professionals in NHCs in New York City. The respondents identified seven problems, including a lack of financial resources, unsafe housing, and emotional distress/depression as affecting large portions of their patient populations. Respondents reported that they are presently meeting many of the psychosocial needs of their clients, but they were pessimistic about their ability to continue to do so due to a lack of funding streams to support their provision of comprehensive health care that includes psychosocial services. The findings suggest that while NHCs may be "holding their own" in providing quality services to their clients, this will be harder to sustain in the future if the numbers of the uninsured served continues to increase, and the revenues generated continue to decrease. PMID- 15810566 TI - Cigarette advertising in magazines for Latinas, White women, and men, 1998--2002: a preliminary investigation. AB - Cigarette ads in popular magazines play a role in smoking and in brand preferences among women and men, but few studies have analyzed ads directed at women vs men, and no study has examined ads directed at women of different ethnic groups. Hence, we examined cigarette ads in popular magazines for White women, Latinas, and men 1998 through 2002 for the first time. Significant differences in the number of cigarette ads by magazine audience were found, along with significant differences in the type and brands of cigarettes advertised to each group. These preliminary findings suggest that the tobacco industry may target women in a manner that differs from its targeting of men, and may target Latinas in a manner that it does not target White women. Results are discussed in terms of the need for further research on tobacco ads directed at women. PMID- 15810567 TI - [Health promotion and prevention in a transition period]. PMID- 15810568 TI - [Overweight and obesity. Social consequences, self-esteem and quality of life, with focus on children and teenagers]. PMID- 15810569 TI - [Peer-to-peer communication as a drug prevention method--seen from a researcher's perspective]. PMID- 15810570 TI - [Discussing menopause in family practice: risk prevention or health promotion?]. PMID- 15810571 TI - [Attitudes and beliefs influencing whether older Danes accept participation in intervention programs for the prevention of falls]. PMID- 15810573 TI - [Transportation and health. Odense - The National Cycling City of Denmark, 1999 2002 ]. PMID- 15810572 TI - [The effect of a health-promoting poster on the choice of whether to take the stairs or the escalator in two S-train stations in Copenhagen]. PMID- 15810574 TI - [The story of the reversal of opinion about hormone replacement therapy and cardiovascular disease]. PMID- 15810575 TI - [Cardiovascular diseases. From epidemiology to prevention]. PMID- 15810576 TI - [Resuscitation with automated external defibrillators outside hospital]. PMID- 15810577 TI - [Prediction of individual residual survival times using statistical models]. PMID- 15810578 TI - [Preventive medicine in Danish hospitals]. PMID- 15810579 TI - [Use of propranolol for migraine prophylaxis]. PMID- 15810581 TI - [Endovascular management of a pseudoaneurysm of the gastroduodenal artery]. PMID- 15810580 TI - [Atkins and other low-carbohydrate diets: hoax or an effective tool for weight loss?--secondary publication]. PMID- 15810582 TI - [Arterial hypertension in autonomous insufficiency]. PMID- 15810583 TI - [Establishment of plantlet regeneration system of medicinal plant Aconitum coreanum]. AB - OBJECTIVE: To set up the rapid propagation and plantlet regeneration system of medicinal plant Aconitum coreanum (Levi.) Rpaics. METHOD: Adventitious buds and regenerated plantlet were obtained by shoot tip culture through different media and combination of plant growth regulators. RESULT: MS medium was the optimal medium for differentiation and proliferation of shoot tip. MS medium with 6-BA 2 to approximately 5 mg/L was found to be most suitable for the induction of bud. MS with 6-BA 2 mg/L + GA3 1 to approximately 5 mg/L or adenine 1 to approximately 5 mg/L was best combination for the growth and proliferation of adventitious buds, and MS with IBA 0.5 to approximately 1 mg/L for the induction of root. CONCLUSION: Tip culture may be used as a useful method to get regenerated plantlet of Aconitum coreanum under certain conditions. PMID- 15810584 TI - [Quality evaluation of Alisma orientalis from different habitats]. AB - OBJECTIVE: To study the seed quality of Alisma orientalis (Sam.) Juzep. METHOD: Collecting seeds from different habitats and studying the thousand-grain weight, moisture content, purity and germination percentage. RESULT: The seeds reaped in summer friom Pengshan, Sichuan province are better than those from other habitats. Its thousand-grain weight is 0.4682 g, moisture content is 12.48%, purity of seeds is 89.18% and germination percentage is above 95%. CONCLUSION: The cause of qualitative differences in seeds of Alisma orientalis from different habitats were analysed in this paper. PMID- 15810585 TI - [Determination of psoralen and isopsoralen in Psoralea corylifolia L. from different habitats]. AB - OBJECTIVE: To study the psoralen and isopsoralen content in Psoralea corylifolia from different habitats. METHOD: C18 column was used. Mobile phase was methanol water (45:55). The detection wavelength was at 245nm and the flow rate was 1.0 ml/min. RESULT: The linearity was obtained over the range of 0.448 to approximately 0.448 microg (r = 0.9999) for psoralen, 0.356 to approximately 0.356 microg = 0.997) for isopsoralen. CONCLUSION: The contents of psoralen and isopsoralen in Psoralea corylifolia from different habitats have no obvious difference. PMID- 15810586 TI - [Content determination of loganic acid and gentiopicroside in Tebitan herbal medicines Gentiana macrophylla and G. straminea by HPLC]. AB - A quantitative method to determine loganic acid and gentiopicroside in Gentiana macrophylla Pall. and Gentiana straminea Maxim. by RP-HPLC was established in this paper. The two compounds were base-isolated on the coloumn of Zorbax SB-C18 (250 mm x 4.6 mm i.d., 5 microm) which eluted with MeOH and H2O (0.04% H3PO4). The methol ratio increased from 18% to 28% in 15 min with detective wave at 254 nm and coloumn temperature at 30 degree C, flow rate at 1 ml/min. A wide range of linearity and the good linear relationships were achieved on above condition. The contents of the two compounds in different parts of the two plants were also determined and compared in this paper. PMID- 15810587 TI - [Study on the microwave extraction and chemical constituents of the essential oil from Amomum tsao-ko in Jinping, Yunnan province]. AB - Essential oil from Amomum tsao-ko collected in Jinping, Yunnan province was obtained by microwave extraction, common solvent extraction and vapor distillation, respectively. Chemical constituents were analyzed by GC-MS and their relative contents were determined by area-normalized method. PMID- 15810588 TI - [Study on chemical constituents of the essential oil from Myristica fragrans Houtt. by supercritical fluid extraction and steam distillation]. AB - Essential oils were extracted from Myristica fragrans Houtt. by supercritical fluid extraction (SFE) and steam distillation (SD). Their components were analyzed by gas chromatography-mass spectrometry and compared. 48 compounds were identified for the essential oil extracted by supercritical carbon dioxide, and its main components have been found to be myristic acid, myristicin, terpinen-4 ol, alpha-pinene and safrole. 38 compounds were identified for the essential oil obtained by SD, and its main components have been found to be beta-pinene, terpinen-4-ol, alpha-pinene, gamma-terpinene and beta-phellandrene. PMID- 15810589 TI - [Determination of coumarins content in Radix Angelicae dahuricae by HPLC and UV]. AB - OBJECTIVE: To determine the coumarins content in Radix Angelicae Dahuricae (Baizhi). METHODS: The total content of coumarins was determined by UV. The content of imperatorin, isoimperatorin and oxypeucedanin hydrate in different samples was carriedut by HPLC. RESULTS: The processing method and the medicinal position have great effect on the coumarin content. CONCLUSION: The content of oxypeucedanin hydrate in Baizhi was determined. This method can be used for quality control of Baizhi. PMID- 15810590 TI - [Purification of chymopapain and its effect on nucleus pulposus in vitro]. AB - OBJECTIVE: To establish a simple and effective procedure for purification of chymopapain and study about its effect on nucleus pulposus in vitro. METHODS: Chymopapain was purified by ion exchange chromatograph and tested its effect on nucleus pulposus in vitro. RESULTS: A simple and effective procedure for purification of chymopapain was established and the chymopapain did degrade nucleus pulposus. CONCLUSION: The ion exchange chromatograph was a simple and effective procedure for purification of chymopapain. It is necessary to study its effect on nucleus pulposus in vivo. PMID- 15810591 TI - [Studies on chromatography fingerprint of huangqi by HPLC]. AB - OBJECTIVE: To study on chromatography fingerprint(CFP) of Huangqi herbs. METHOD: HPLC was used on a Hypersil ODS column with water-acetonitriles gradient elution. CFP was referenced to formononetin, calycosin and then the contents of astragaloside IV, formononetin and calycosin was determinted. RESULT: 26 common peaks were pointed out, their similarity of 12 samples were more than 90%. CONCLUSION: The handling of samples were simple. The study method on chromatography fingerprint of Huangqi were reliable. Good repetition and their similarity accorded with the demand on "the technology guide to experiment studies on chromatography fingerprint of injection of traditional Chinese medicine (TCM)", which was issued by the nation pharmacopoeial commission. The chromatography fingerprint and determination of index composition could be used to comprehensively control the quality of Huangqi. PMID- 15810592 TI - [The effect of matrine on the expression of P170 LRP and TOPO II of obtained multi-drug resistance of mouse S180's tumour cell]. AB - OBJECTIVE: To observe the effect of matrine on P170 expressed by multi-drug resistance gene, lung resistant protein (LRP), and boposomeras II. METHODS: By the method of less dosage of chemotherapy PFC, the mouse model of multi-drug resistance of S180 tumour cell was set up. At the same time, matrine was administered to the mouse model for 4 weeks, and then P170 LRP, TOPO II were observed by flow cytometry. RESULTS: Martine can obviously reduce the express of P170, LRP and the activity of TOPO II in multi-drug resistance tumour cell induced by chemotherapy. CONCLUSION: Matrine can intervene the occurrence of the multi-drug resitance of tumor cells induced by chemotherapy. PMID- 15810593 TI - [Purification of small peptide from Agkistrodon blomhoffii Bioe and its inhibition on platelet aggregation]. AB - This is the first report to purify a new peptide SV-PP-1 from Agkistrodon blomhoffii Bioe. SV-PP-1 has molecular weight of 1234.616 Da and can inhibit on platelet aggregation. SV-PP-1 significantly inhibited the ADP-induced platelet aggregation. It has been found up to now that SV-PP-1 is the smallest peptide inhibiting on platelet aggregation from snake venom. PMID- 15810594 TI - [Anti-tumor effects of ampelopsin on human lung cancer GLC-82 implanted in nude mice]. AB - OBJECTIVE: To investigate the effects of ampelopsin on the growth of human lung cancer cell line GLC-82 in nude mice. METHODS: GLC-82 cells were inoculated into the armpit of nude mice to establish lung cancer model. Then the BALB/C nude mice 6 with xenograft tumor were randomized into 6 groups. Ampelopsin was administered at 3 dosages by intraperitoneal injection daily. The anti-tumor effect of ampelopsin was evaluated on tumor volume, relative tumor volume, tumor weight, relative tumor proliferative rale, and tumor growth curve. RESULTS: At the 250 mg/kg dose of ampelopsin, the growth inhibition of the transplant tumor was 35.5% (P < 0.01) and 37.1% (P < 0.01), and the relative tumor proliferative rate was 55.24% (P < 0.01) and 57.71% (P < 0.05), respectively in two independent experiments by compared with the normal saline control. CONCLUSION: Ampelopsin appears to be a potent anti-tumor agent that was firstly discovered in the transplant human lung cancer cell line GLC-82 in nude mice in vivo. PMID- 15810595 TI - [Effect of fractions isolated from Naja naja atra venom on antioxidation in animals]. AB - OBJECTIVE: To study the antioxidation of fraction F and H isolated from Naja naja atra venom on homogenate and RBC autoxidation. And to explore the effect of two fractions on the activities of antioxidation enzymes in mice. METHOD: Samples of tissues homogenates and RBC suspension were pretreated with fraction F/H, and then their generation of malondialdehyde(MDA) was examined. The effects of fraction F/H on O2-* produced by pyrogallol autoxidation and *OH produced by Cu2+ -VitC were checked. After being administrated i.p. with fraction F/H for 15 days, superoxide dismutase and hydroperoxidase in mice were measured for their activities. RESULT: Fraction F/H inhibited RBC autohemolysis and the generation of MDA decreased. The production of lipid peroxides in normal brain, liver and heart homogenate of rat and the elevation of lipid peroxides induced by cysteine and FeSO4 in homogenate were inhibited by addition of fraction F/H. The O2-* produced by autoxidation of pyrogallol and the *OH produced by Cu2+ -VitC system could be scavenged by fraction F/H (the potency of scavenging oxygen free radical is that fraction H > fraction F). Fraction F/H all increased the activities of several antioxidation enzymes in mice to different extents. CONCLUSION: Fraction F/H have the effect of antilipid peroxidation,may scavenge active oxygen free radical and increase the activity of SOD. PMID- 15810597 TI - [Effects of ethanol extracts from Triptergium wilfordii hook and Stephania tetrandra S. Moore on lymphocytes and cyclooxygenase]. AB - OBJECTIVE: To explore the effects of extracts from Triptergium wilfordii Hook and Stephania tetrandra S. Moore on lymphocytes and cyclooxygenase. METHODS: Change in proliferation of mice thymic lymphocytes and spleen lymphocytes, contents of 6 keto-PGF1alpha in bovine arterial endothelial cells, PGE2 in murine macrophages were determined. RESULTS: Proliferation of mice thymic lymphocytes and spleen lymphocytes were inhibited by alcoholic extracts from Triptergium wilfordii Hook, Stephania tertrandra S. Moore. COX-1 and COX-2 were inhibited to different extent by ethanol extracts from Triptergium wilfordii Hook. COX-1 was inhibited by ethanol extracts from Stephania tetrandra S. Moore. CONCLUSIONS: Part of anti rheumatic mechanism of the two herbs might probably be mediating through inhibition of lymphocyte proliferation, COX-1 and COX-2. PMID- 15810596 TI - [The effect of curcumin on bladder cancer cell line EJ in vitro]. AB - OBJECTIVE: To observe the effect of curcumin on bladder cancer cell line EJ in vitro. METHODS: Cell morphology, MTT, flow cytometer, immunocytochemical method for detecting NF-KB, Cyclin D1 were used to observe the effect of 5,10,20 mg/L curcumin on bladder cancer cell line EJ in vitro. RESULTS: All concentrations curcumin resulted in the growth suppression significantly [Suppression ratio > or = (27.5 + 3.1)%, P < 0.05]. Above 10 mg/L concentrations curcumin induced apoptosis [Apoptosis ratio > or = (14.6 +/- 1.8)%, P < 0.05] and down-regulated of the expression of NF-kappaB [Expression ratio < or = (35.8 +/- 4.2)%, P < 0.05], Cyclin D1 [Expression ratio < or = (29.7 +/- 3.2)%, P < 0.05]. The cell phase arrest induced by curcumin was G1 phase arrest mainly with significant decrease of S phase. CONCLUSIONS: Curcumin can suppress the growth, induce apoptosis of bladder cancer EJ cell in vitro. Its mechanism is related with down regulations of the expressions of NF-kappaB and Cyclin D1. Curcumin has great potential for the treatment of bladder cancer. PMID- 15810598 TI - [Effect of weiluo tongmai capsule on hemorrheology in rats]. AB - OBJECTIVE: To observe the effects of Weiluo Tongmai Capsule on hemorrheology. METHODS: Several items of hemorrheology were assayed, including specific viscosity of whole blood and plasma, erytlrocyte sedimentation rate, erythrocyte hematocrit, thrombosis in vitro in normal and blood stagnation rats. RESULTS: Weiluo Tongmai Capsule significantly lowered specific viscosity of whole blood, markedly inhibited dry weight and wet weight of thrombus in vitro, and tended to inhibit thrombus length and erythrocyte sedimentation rate in blood stagnation rats induced by adrenaline and ice water. CONCLUSION: Weiluo Tongmai Capsule had the effects on improving items of hemorrheology and activation blood. PMID- 15810599 TI - [Study on Enrichment process of total flavones from Semen cuscutae with macroporous resin]. AB - OBJECTIVE: To study the optimal conditions and parameters for the enrichment process of total flavones in Semen cuscutae with macroporous resin. METHODS: With the enrichment degree of total flavones in Semen cuscutae as an index, the optimal conditions of the enrichment process were investigated. RESULTS: The aqueous extract from Semen cuscutae was taken into the macroporous resin column, then the resin was washed by distilled water to get rid of impurity and 30% and 70% ethannol to elute total flavones. The above conditions were optimal. CONCLUSIONS: The process is feasible to enrich effective ingredient flavones from Semen cuscutae. PMID- 15810600 TI - [Microwave-assisted extraction technology of skin-lightning agent in Rhizoma Typhonii]. AB - The microwave-assisted extraction (MAE) technology of skin-lightning agent in Rhizoma Typhonii was studied and the relative inhibitory activity of the extract against tyrosinase was used as evaluating criterion. It was concluded that the optimal condition was two doses of microwave radiation (450 W) for 7 min with a mass ratio of water to material of 10: 1. The dry extract of Rhizoma Typhonii with 20.9% yield and 47.1% relative inhibitory activity against tyrosinase was obtained. Compared with direct-heating extraction, MAE can shorten extracting duration, and improve relative inhibitory activity of the extract against tyrosinase. PMID- 15810601 TI - [Study on preparation of beta-cyclodextrin inclusion volatile oil from Rhizoma Curcumae]. AB - OBJECTIVE: To study optimum inclusion process conditions for volatile oil from Rhizoma Curcumae. METHOD: The study was carried out with orthogonal design. The process conditions were studied by determining the utilazision ratio of volatile oil from Rhizoma Curcumae, the oil-bearing rate and extract ratio of inclusion compound. RESULTS: The optimum preparation conditions for inclusion were established as: volatile oil:beta-CD was 1:8, the inclusion temperature was at 80 degrees C and inclusion time for 3 h, ultilizasion rate of volatile oil was 86%. CONCLUSION: The method can be used for mass production. PMID- 15810602 TI - [Study on extraction process of the volatile oil for biyuan dropping pills]. AB - OBJECTIVE: To determine the optimal extracting process of the volatile oil for Biyuan Dropping Pills. METHODS: Conditions for the extraction were studied by orthogonal design guided by the increasement of the volatile oil. RESULTS: The distilling time is the main factor,and the fineness of the herbal material is the second. The optimal process is as follows: crush the herbal material into coarse powder, macerate 1 hour with 9 times water and distill 4 hours. CONCLUSION: The optimized process is feasible and stable. PMID- 15810603 TI - [Clinical study of zhuyutongmai capsule on treating senile cerebral infarction and effect on function of vascular endothelium]. AB - OBJECTIVE: To evaluate the clinical study of Zhuyutongmai (ZYTM) Capsule in treating senile cerebral infarction and effect on function of vascular endothelium. METHODS: Sixty-four aged patients with cerebral infarction were randomly divided into the treated group treated with ZYTM and the control group treaded with Naoluotong. The clinical effect was observed. The hemorrheologic and hemodynamic parameters, the blood levels of endothelin (ET) and nitric oxide (NO) were determined before and after treatment. RESULTS: After treatment of ZYTM, the hemorrheologic and hemodynamic parameters were all significantly improved (P < 0.01 or P < 0.05), the therapeutic effect of ZYTM was better than that of Naoluotong (P < 0.05), compared with the control group after treatment, the level of ET in the treated group were lower (P < 0.01), and level of NO were higher (P < 0.01). CONCLUSION: ZYTM has good effect in treating senile cerebral infarction. PMID- 15810604 TI - [Structural, functional and genetic aspects of peroxisome biogenesis]. AB - Intracellular organelles, peroxisomes, occur in cells of most eukaryotic species. Human severe congenital disorders are associated with defective assembly and functioning of peroxisomes, which partly explains the attention of researchers paid to peroxisome biogenesis. It has been shown that peroxisomes are involved in the realization of eukaryotic developmental programs (in particular, neuroblast differentiation and postembryonic development). Cytobiochemical and electron microscopic studies of mutations involving peroxisomes showed that the primary role in peroxisome biogenesis is played by synthesis of proteins (peroxins) and their transport and incorporation into peroxisome membranes. More than 30 peroxin encoding genes have been examined. These genes are synthesized on free polysomes and transported into peroxisomes by means of specific signaling peptides, PTS1, PTS2, and PTS3. The import of matrix proteins depends on at least two shuttle receptor proteins, Pex5p and Pex7p. Some proteins regulating peroxisome proliferation in cells have been identified. PMID- 15810605 TI - [Identification of the gene whose mutation leads to the morphological changes in the hypocotyl of Arabidopsis thaliana]. AB - The results of genetic and molecular genetic analysis of line 176 of Arabidopsis thaliana with reduced hypocotyls obtained from a previously obtained collection of insertion mutants, are presented. The examined mutation proved to be recessive and based on a single insertion of the T-DNA vector pLD3 into the A. thaliana genome. Computer-aided analysis of the DNA region adjacent to the left border of the insertion revealed a putative site of T-DNA insertion, the 2.5-kb At2g09920 gene located in the long arm of chromosome 2, near the centromere. PMID- 15810607 TI - [Association between defects of karyogamy and translation termination in yeast Saccharomyces cerevisiae]. AB - Mutants capable of a high frequency of cytoduction (Hfc+) were obtained in a haploid strain of Saccharomyces cerevisiae, suggesting impaired cytogamy. Nine of the 68 Hfc+ mutants showed the antisuppressor effect with respect to mutations of the SUP35 and SUP45 genes, which code for translation termination factors, or to the [PSI+] factor, which is the prion form of Sup35. Cosegregation of the characters "higher frequency of cytoduction" and "antisuppression" was demonstrated for three Hfc+ mutants. One (HFC12-2) of the Hfc+ mutations exerted a dominant antisuppressor effect with respect to [PSI+] and had no effect on [PSI+] maintenance. On the strength of the results, an interaction was assumed for translation termination components and cytoskeleton proteins, which play a role in karyogamy in yeasts. PMID- 15810606 TI - [Comparative study of the expression of the native and modified cry3a genes of Bacillus thuringiensis in prokaryotic and eukaryotic cells]. AB - The cry3a gene of Bacillus thuriengiensis was cloned. Based on sequence analysis of this gene, a modified gene, cry3aM, was constructed, which has the optimal codon composition for effective expression in eukaryotic cells. Hybrid genes cry3a-licBM2 and cry3aM-licBM2 were constructed, in which the sequences of the native and modified genes are fusedfused with the reorter gene for thermostable lichenase in the reading frame. We have shown that the expression levels of hybrid genes cry3a-licBM2 and cry3aM-licBM2 in Escherichia coli are comparable, being 5% of those for reporter gene licBM2. In cells of a lower eukaryote Saccharomyces cerevisiae, the expression of hybrid gene cry3aM-licBM2? Which contains the modified gene, considerably exceeded the level of expression of cry3a-licBM2 containing the native gene. The presence of lichenase in the composition of hybrid proteins was shown to facilitate selection and analysis of the expression level of hybrid proteins in transgenic organisms. PMID- 15810608 TI - [Divergence of the polytene chromosome banding sequences as a reflection of evolutionary rearrangements of the genome linear structure]. AB - Banding sequences of five chromosomal arms (A, C, D, E, and F), accounting for about 70% of the total genome size in 63 Chironomus species, were used as phylogenetic markers to analyze divergence patterns of the linear genome structure during the evolution. The number of chromosomal breakpoints between the pairs of banding sequences compared served as a measure of divergence. It was demonstrated that the greater the divergence between the species compared, the higher the number of chromosomal breakpoints and the smaller the size of the conserved chromosomal regions. A banding sequences comparison in sibling species demonstrated a lower number of chromosomal breakpoints; the breakpoint number was maximum in a comparison of the banding sequences in the subgenera Chironomus and Camptochironomus. The use of the number of chromosomal breakpoints as a divergence measure provided establishment of phylogenetic relationships between 63 Chironomus species and discrimination of sibling species groups and cytocomplexes on a phylogenetic tree. PMID- 15810609 TI - [Overexpression of a novel gene toothrin in Drosophila]. AB - A newly found locus of the Drosophila melanogaster genome, named toothrin (tth) has been used to study the role of the conserved domain 2/3 of genes from the d4 family. In contrast to the 2/3 domain of all vertebrates studied (including humans), which is always accompanied by the d4 domain, the tth gene contains the sequence encoding the 2/3 domain but lacks that encoding the d4 domain. The tth gene overexpression has been studied using the two-component system UAS-GAL4. It has been demonstrated that the tth overexpression at the third-instar larval (prepupal) stage decreases survival rate, simultaneously causing a substantial deceleration of development in Drosophila. It is known that the change of developmental stages in Drosophila is controlled by the rates of the expression of ecdysteroid and juvenile hormones (JHs). It is supposed that the overexpression of the tth gene causes either a shift in the ecdysterone-to-JH ratio (through a decreased JH decay rate or a delayed initiation of ecdysone synthesis) or a deceleration of the release of ecdysterones synthesized. PMID- 15810610 TI - [Allozyme variation in sexual and apomictic Taraxacum and Pilosella (Asteraceae) populations]. AB - Allozyme spectra of peroxidase, esterase, superoxid dismutase, tyrosinase, alcohol dehydrogenase, lactate dehydrogenase, and acid phosphatase were examined in populations of sexual (Taraxacum serotinum and Pilosella echioides) and apomictic (T. officinalis and P. officinarum) plant species. The heterozygosity in these populations (0.455-0.620) proved to be considerably higher than the average level characteristic of plant populations (0.058-0.185). The populations examined did not differ in the mean phenotype number mu, i.e., they exhibited the same diversity (3.213-3.380). The proportion of rare phenotypes h also did not differ between the sexual and apomictic species of the same genus, whereas this parameter in the Pilosella populations (0.150-0.174) was significantly higher than in the Taraxacum ones (0.093-0.114). The populations were characterized by numerous isozyme spectra (more than 11 per populations) and displayed multiple allelism (the mean allele frequency was 3.63-4.38 per locus). They exhibited a high percentage of rare (occurring at a frequency lower than 5%) spectra (35 80%). This indicates that agamic complexes, to which these populations belong, may have a more complicated genetic structure of both apomictic and sexual populations than the species that do not belong to agamic complexes. PMID- 15810611 TI - [Population-genetic variation in Scots pine Pinus sylvestris L. from the main forest regions of Ukraine]. AB - Using electrophoretic analysis of 22 isozyme loci controlling ten enzyme systems, we studied intrapopulation and interpopulation variation of Scots pine Pinus sylvestris L. in the main forest regions of Ukraine. In 15 of the populations examined, 76.5% of genes were polymorphic, and an average plant was shown to be heterozygous at 23.4% of the genes. The lowest and highest values of major polymorphism parameters were characteristic of respectively the relic populations of Ukrainian Carpathians and the populations from the steppe and forest-steppe zones. Nei's genetic distances between the populations varied from 0.006 to 0.031 (on average 0.016). Cluster analysis failed to show clear trends in the population distribution relative to their geographical position. PMID- 15810612 TI - [Association of DNA polymorphisms of the growth hormone and prolactin genes with milk productivity in Yaroslavl and black-and-white cattle]. AB - Polymorphisms of the prolactin (bPRL) and growth hormone (bGH) genes were studied comparatively in the Russian and German Black-and-White and Yaroslavl cattle breeds. Two polymorphisms were studied for each gene. In the case of the bPRL gene, the polymorphism of the 5'-untranslated region was examined by microsatellite analysis and the RsaI polymorphism of exon 3, by RFLP analysis. In the case of the bGH gene, the MspI polymorphism of intron III and the AluI polymorphism of exon 5 were assessed by RFLP analysis. Differences in allele and genotype frequencies were observed both between and within breeds. The heterozygosity at the RsaI marker was low (9.4%) in the Russian Black-and-White breed; that at the microsatellite of the bPRL gene was low (3.2-24%) in all breeds examined. Homozygotes BB at the bPRL gene, which had not been reported earlier for European cattle breeds, were detected in the German Black-and-White and Yaroslavl breeds (at frequencies 0.16 and 0.13, respectively). The frequency of allele MspI(-) of the bGH gene in the Yaroslavl breed was extremely low (0.02), comparable only with that of the Holstein cattle (0.02). The heterozygosity at the AluI polymorphism was higher than at the MspI polymorphism of the bGH gene and reached 55% in the Yaroslavl breed. Genotype BB of the RsaI polymorphism of the bPRL gene tended to show a negative association with the fat content in milk. The genotypes of the AluI polymorphism of the bGH gene were associated with the fat content in milk in the Yaroslavl (F = 4.56, P = 0.013) and German Black-and-White (F = 4.1, P = 0.014) breeds: the highest fat content in milk was observed in the subsample of cows with heterozygous genotype VL. PMID- 15810613 TI - [Modeling effect of homo- or heterozygosity on animal growth intensity]. AB - A model has been developed that describes the dependence of a quantitative selective trait on the animal homo- and heterozygosity for the genes that control the biochemical reaction rate (isoenzyme systems). The model includes any cases of gene control of a quantitative trait and is applicable to real, genetically heterogeneous populations. The results of testing the model show that it can be used for identifying the genes involved in additive polygenic determination of quantitative commercially valuable traits. PMID- 15810614 TI - [Population analysis of electrophoretic variation in blood serum albumins of European (Acipencer ruthensis L.) and Siberian (A. ruthensis marsiglii Brandt) sterlet]. AB - We studied blood serum albumins in European (Acipencer ruthensis L.) and Siberian (A. ruthensis marsiglii Brandt) sterlet using disk electrophoresis in polyacrylamide gel. The albumins were shown to be controlled by three codominant alleles of a single locus (ALB*a, b, c). In European sterlet, all three theoretically possible genotypes were described, one of which (ALB*c/c) occurred extremely rarely (one individual). Siberian sterlet was found to be monomorphic for albumins: all fish examined had the ALB*a/a genotype. There was no correlation between albumin patterns and fish fatness. In a number of samples from the Volga River basin, spatial and temporal differentiation was found and analyzed. The results suggest that construction of hydroelectric plants may provoke massive and prolonged starlet migrations. PMID- 15810615 TI - [Genetic and demographic characteristics of rural populations of Altai republic: sex-age composition, surname and tribal structure]. AB - The sex, age, tribal, and surname compositions of the populations of three villages of Altai Republic, Kulada (Ongudaisk raion), Beshpeltir (Chemal raion), and Kurmach-Baigol (Turochak raion) have been studied. Altaian populations are characterized by a high proportion of persons under 20 years of age (35.3-46.1%); however, there is a tendency towards a narrow base of the sex-age pyramid. The sex ratios in the total populations and in individual age groups are unfavorable. The rural populations studied differ in the spectrum and pattern of surname accumulation. The Kurmach-Baigol population (which consists of Northern Altaians) considerably differs from the Beshpeltir and Kulada populations (which are mostly Southern Altaian) with respect to the calculated parameters characterizing the population structure (random isonymy, migration index, the parameter of tribe diversity, entropy, and the redundancy of surname distribution). Isonymy coefficients of relationship between individual populations have been calculated from the data on tribes (surnames). These coefficients for pairs of populations are the following: for the Beshpeltir and Kulada populations, 0.3035938 (0.0000443 and 0.0000378 for the Altaian and total populations, respectively); for the Beshpeltir and Kurmach-Baigol populations, 0.0026788 (0.0000172 and 0.0000121 for the Altaian and total populations, respectively); and for the Kulada and Kurmach-Baigol populations, 0.0054811 (no common surnames have been found). PMID- 15810616 TI - [Genetic and demographic characteristics of rural populations of Altai republic: dynamics of the marriage structure]. AB - The dynamics of population marriage structure in the period from 1951 to 1997 has been studied in three villages of Altai Republic: Kulada (Ongudaisk raion), Beshpeltir (Chemal raion), and Kurmach-Baigol (Turochak raion). These populations have been found to differ from one another in the intensity and direction of migration, as well a its temporal pattern with respect to the birthplaces and ethnicity of persons contracting marriages. Periods of active incorporation of non-Altaian (mostly Slavic) ethnic components into the gene pools of the Beshpeltir and Kurmach-Baigol populations have been detected. The geographic distributions of the birthplaces of men and women contracting marriages are different. Therefore, migration has different effects on the genetic diversity of the Y-chromosomal and mitochondrial-DNA pools. No isonymic marriages have been found in the Altaian populations studied; however, an increase in the random component of inbreeding has been observed. PMID- 15810617 TI - [Kohonen network study of the results of RAPD and ISSR analyses of genomic polymorphism in the genus Capsicum L]. AB - The results of studies based on multilocus molecular analyses, including random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and amplified fragment length polymorphism (AFLP) analyses, are usually presented in the form of images (electrophoregrams, photographs, etc.). The interpretation of this information is complicated, labor-consuming, and subjective. Artificial neural networks (ANNs), which are ideal "image processors," may be useful when solving such tasks. The possibility of using ANNs for the treatment of the results of RAPD and ISSR analyses has been studied. The RAPD and ISSR spectra have been studied in fragments of DNA of plants from the genus Capsicum L. (peppers). The results of clustering the accessions studied by means of the unweighted pair-group method with arithmetic averages (UPGMA), which is often used for phylogenetic constructions based on RAPD and ISSR data, serve as expert estimates. Fundamentally new methods of genetic polymorphism estimation using ANN technologies, namely, self-organizing feature maps (SOFMs) have been developed. The results show that the clusters obtained with the use of UPGMA and SOFM coincide by more than 90%; taking into account that ANNs can deal with high noise levels and incomplete or contradictory data, the approach proposed may prove to be efficient. PMID- 15810618 TI - Variation in survival of Iris pumila L. juvenile plants grown in two nutrient regimes. AB - I analyzed variability of early life survival on 881 Iris pumila juvenile plants in their prereproductive period. Seedlings were produced by applying half-sib mating scheme on clones originating from two Deliblato Sand populations that were accommodated to common garden conditions. Nineteen clones served as pollen donors (sires), and the other 90 clones served as pollen recipients (dames). Seedlings were grown in two nutrient levels (full strength and 1/10 of full-strength Hoagland solution). While I failed to detect significant mean nutrient level effect as well as significant between population differentiation for prereproductive survival, a statistically significant additive genetic variability for this ultimate prereproductive measure of fitness was detected. PMID- 15810619 TI - How to make a Barrett esophagus: pathophysiology of columnar metaplasia of the esophagus. AB - Barrett esophagus is defined as a specialized intestinal replacing the squamous epithelium of the esophageal mucosa in response to gastroesophageal reflux. Barrett metaplasia is a healing process that develops to protect the esophagus from further damage. Although mechanisms by which Barrett metaplasia evolves toward dysplasia and adenocarcinoma have been extensively studied, the process by which squamous epithelium is replaced by specialized intestinal metaplasia is poorly understood. Barrett esophagus develops when defense mechanisms in the esophageal mucosa (luminal secretion of mucus, bicarbonate, growth factors, etc.) are overwhelmed by an ongoing cycle of mucosal injury and repair. Hydrogen ion, pepsin, trypsin, and bile acids are considered harmful agents that synergistically invade the esophageal mucosa. Areas of destroyed squamous epithelium are then progressively reepithelized by a columnar epithelium that may originate from multipotent stem cells located within the basal layer of the normal esophageal mucosa or in the ducts of submucosal glands. PMID- 15810620 TI - Expression of Cdx2 in early GRCL of Barrett's esophagus induced in rats by duodenal reflux. AB - The intestine-specific caudal-related homeobox transcription factor Cdx2 is widely accepted to play a key role in intestinal development and differentiation in mammals. We studied the role of Cdx2 in the development of Barrett's esophagus (BE). In previous studies, we have shown a sequence of morphological changes of squamous epithelium leading to BE, found a peculiar metaplastic change common to other parts of gut, and proposed the concept of a "gut regenerative cell lineage" (GRCL). The GRCL is characterized by pyloric-foveolar metaplasia with goblet cell metaplasia, which occurs in the regenerative process in response to chronic inflammation. BE very likely develops through the GRCL, and we studied the expression of Cdx2 in various lesions of rat esophageal mucosa induced by duodenal reflux, using reverse transciptase-polymerase chain reaction and immunohistochemistry against Cdx2. We found that Cdx2 was expressed not only in specialized columnar epithelium (SCE) of BE, but also in several pyloric gland and foveolar metaplastic cells which developed in the basal layer of the squamous epithelium at an earlier stage of SCE development. These findings indicate that Cdx2 plays a crucial role in directing intestinal-type differentiation of the GRCL. PMID- 15810621 TI - Helicobacter pylori infection in patients with erosive esophagitis is associated with rapid heartburn relief and lack of relapse after treatment with pantoprazole. AB - Previous studies have shown an increased effect of proton pump inhibitors on intragastric pH in Helicobacter pylori (HP)-infected patients suffering from gastroesophageal reflux disease (GERD). We evaluated the effect of HP infection on healing and symptom relief in GERD patients with erosive esophagitis treated with pantoprazole. Two hundred twenty-seven patients with endoscopically proven reflux esophagitis were treated for 8 weeks with pantoprazole, 40 mg daily. Patients achieving endoscopic healing at that time were treated for 16 weeks more with pantoprazole, 20 mg daily. Healing and symptom relief rates for HP infection were compared. We found complete relief of heartburn in 72.3% of the HP-positive versus 58.8% of the HP-negative group (P < 0.05). Overall prevalence of heartburn at 8 weeks was higher in the HP-negative group (40.3 vs. 25.8%; P < 0.05), with no significant differences in endoscopic healing (overall 80.4%). At 24 weeks of treatment, the symptomatic relapse rate was higher in the HP-negative group (25.9 vs. 10.2%; P < 0.020). At 8 weeks, patients with erosive esophagitis and HP infection exhibited a significantly better response to pantoprazole through complete heartburn relief, with no differences in endoscopic healing rates between the groups. After 24 weeks, the relapse rate was significantly higher in the HP-negative group. PMID- 15810622 TI - Triple-a syndrome: a rare etiology of adult achalasia. PMID- 15810623 TI - Twice-daily standard dose of omeprazole achieves the necessary level of acid inhibition for Helicobacter pylori eradication. A randomized controlled trial using standard and double doses of omeprazole in triple therapy. AB - Antibiotic resistances and level of acid inhibition may affect the outcome of eradicating regimens for H. pylori. To evaluate the impact of different degrees of acid inhibition on the efficacy of triple treatment, we treated 323 patients with H. pylori infection with clarithromycin and tinidazole plus omeprazole, either 20 mg bid or 40 mg bid. Gastric biopsies and antimicrobial susceptibility testing were performed. Eradication was evaluated by means of breath test. Eradication rates were (intention to treat and per protocol) 83.3 and 84.3% in patients receiving 40 mg omeprazole and 81.9 and 84.1% in those receiving 80 mg omeprazole. Culture was successful in 218 patients (68.7%). Resistance to clarithromycin and metronidazole were found in 13.7 and 20.6%, respectively. Eighteen further patients (8.2%) presented double resistance. Resistance was comparable across the two groups. In resistant patients the eradication rate was significantly lower (66.6% [95% CI, 56-76%], vs 86% [95% CI, 78-91%]; P = 0.001). Antibiotic resistance (OR, 2.73; 95% CI, 1.4-5.3) and smoking (OR, 2.68; 95% CI, 1.4-5.2) were independent predictors of eradication failure. Omeprazole, 20 mg bid, achieves the optimal acid inhibition in H. pylori eradication. Increasing antisecretory activity does not significantly enhance cure rates. PMID- 15810624 TI - Comparison of the clinical feasibility of three rapid urease tests in the diagnosis of Helicobacter pylori infection. AB - Rapid urease tests (RUTs) are a fast, accurate, and inexpensive method to diagnose H. pylori infection in the endoscopy suite. Of these, the CLO test is both common and widely used. The aim of our study was to evaluate the accuracy and positive reaction times of two new rapid urease tests (ProntoDry and HpONE) in comparison with the CLO test. Fifty-one patients (26 men, 25 women; mean age, 52.4 years) were included in this study, and all underwent esophagogastroduodenoscopy (EGD). None of the patients had received any prior H. pylori eradication therapy. H. pylori infection status was evaluated by histology, culture, 13C-UBT, and RUT. H. pylori infection was considered to be positive if the culture was positive or if two of the other three tests (histology, RUT, and 13C-UBT) were positive. If culture was negative and only one of the other three tests was positive, or if all four tests were negative, the result was interpreted as negative. Of these 51 patients, 2 were excluded and 29 (59.1%) were infected with H. pylori. The sensitivities, specificities, positive predictive values, and negative predictive values of the three RUTs were not significantly different. The mean positive reaction times of the three RUTs (CLO test, ProntoDry, and HpONE) were 67.8+/-12.0, 16.5+/-2.2, and 17.8+/-2.1 min, respectively. ProntoDry (P < 0.001) and HpONE (P < 0.001) had significantly faster reaction times than the CLO test, but there was no significant difference between ProntoDry and HpONE. Different media of RUTs may influence the rapidity of a positive reaction time. Both ProntoDry and HpONE were superior to the CLO test in terms of accuracy, reaction time, and cost-effectiveness. PMID- 15810625 TI - Evaluation of the stool antigen test for Helicobacter pylori in children and adolescents. AB - The stool antigen test for Helicobacter pylori is a noninvasive immunoassay to diagnose active infection with Helicobacter pylori in human fecal samples. Its performance in children and teenagers has been tested in some developed countries, showing a sensitivity and specificity above 90%, however, its accuracy in developing countries and in children under 6 years is not well established. To determine the accuracy of the test for diagnosing Helicobacter pylori infection in children and teenagers, we evaluated 133 patients (4 months to 17 years old). The gold standard was a positive culture or positive histology and rapid urease test. The test was done according to the manufacturer's instructions. However, modifications were introduced for better reproducibility. Samples were analyzed twice and results are expressed as optical density (OD) determined spectrophotometrically at 450 nm. HpSA was considered positive at OD > or =0.160 and negative at OD <0.140. One hundred twenty-seven of the 133 (95.5%) patients were included. There were no infected infants. The test showed a 94.6% sensitivity (95% CI: 90.6-98.5) and a 96.5% specificity (95% CI: 93.3-99.7). At ages 2 to 6 years the specificity was 96.4% (95% CI: 85.1-99.2) and the sensitivity was 80.0% (95% CI: 64.8-89.7), at ages 6 to 10 years the sensitivity was 100.0% and the specificity 95.7%, and above 10 years the sensitivity and specificity were 100.0%. We conclude that the test is efficient in adolescents and children, however there is a need for further studies with a greater number of patients for evaluation of its accuracy in infants. PMID- 15810626 TI - Endoscopic evaluation of gastrointestinal lesions noted incidentally on PET scanning. PMID- 15810627 TI - Degradation of polyurethane gastrostomy devices: what is the role of fungal colonization? AB - The aim of this study was to evaluate polyurethane percutaneous endoscopic gastrostomy (PEG) tube degradation and the role played by fungi. The inner surfaces of 20 used polyurethane tubes were brushed, and the brushing end was incubated for 7 days in Saburaud broth and cultured if fungal growth occurred. Three tubes used for 12 (sample 12w), 17 (sample 17w), and 96 (sample 96w) weeks and two new tubes were cut to produce several 4-cm-long equal halves. Six samples from the new tubes were considered control samples (Co sample), seven were incubated in Saburaud broth (Co sample + Sa.), and seven in the broth supplemented with Candida albicans (Co sample + Sa. + Ca). All samples underwent morphological examination by electron microscopy and differential scanning calorimetry measurements (DSC). All tubes had fungal colonization. DSC showed deterioration in all tubes including the new ones; adding Candida albicans had no additional effects. Morphological examination by electron microscopy showed a regular pattern in the Co sample, and thick biofilm, holes, and crevices in samples 12w, 17w, and 96w. The more the tubes had been used, the more severe were the changes. The Co sample + Sa and the Co sample + Sa + Ca showed no changes in the inner surface, but cryogenically fractured surfaces had holes and crevices. Yeasts constantly colonize PEG tubes and are likely to contribute to polyurethane deterioration. The impairment of new PEG tubes incubated in Saburaud broth suggests that other factors also play a role in polyurethane deterioration. PMID- 15810628 TI - Gastric outlet obstruction caused by gallstones: case report and review of the literature. PMID- 15810629 TI - Biomarkers in various types of atrophic gastritis and their diagnostic usefulness. AB - Atrophic gastritis has been shown to involve either the oxyntic gland area, resulting in hypergastrinemia and hypopepsinogenemia I, the antral gland area, causing hypogastrinemia without change in serum pepsinogen I (diffuse antral gastritis; DAG), or the entire gastric mucosa (multifocal atrophic gastritis; MAG), resulting in both hypogastrinemia and hypopepsinogenemia I; and rare atrophic gastritis limited to the oxyntic gland area, with antibodies against oxyntic cells and/or intrinsic factor (autoimmune metaplastic atrophic gastritis; AMAG). This study was performed on 126 patients with various forms of gastritis and on 126 age- and gender-matched controls, who were subjected to endoscopy with biopsy, H. pylori testing (13C-UBT, serology), assays for serum gastrin and pepsinogen I, and testing for basal and pentagastrin-induced gastric acid secretion. The following groups of patients were examined: group I (N = 22), with AMAG; group II (N = 53), with DAG; group III (N = 51), with MAG; and group IV (N = 126), age- and gender-matched controls without gastritis. The following changes were found. In group I very high serum gastrin and very low pepsinogen I were observed, and all patients were achlorhydric and H. pylori negative. In group II, with low serum gastrin and normal pepsinogenemia and gastric chlorhydria, all patients were H. pylori positive. In group III, with lower serum gastrin and lower pepsinogen I levels and reduced chlorhydria, all patients were also H. pylori positive. And all group IV controls, with normal serum gastrin and pepsinogen I and normal gastric acid secretion without antral or fundic gastritis, were H. pylori negative. We conclude that measurements of serum gastrin and pepsinogen I and gastric acid secretion as well as testing for H. pylori infection may be useful in noninvasive diagnosis of various types of atrophic gastritis and in identification of patients with premalignant gastritis and a high risk of gastric cancerogenesis. PMID- 15810630 TI - Association between 6-n-propylthiouracil (PROP) bitterness and colonic neoplasms. AB - Inadequate vegetable intake appears to increase colon cancer risk. Since genetic variation in taste influences vegetable preference, we tested associations between bitterness of 6-n-propylthiouracil (PROP), a measure of taste genetics, and number of colonic polyps, a measure of colon cancer risk, in 251 men who underwent screening lower endoscopy. Patients used the general Labeled Magnitude Scale to rate bitterness of 1.6 mg PROP delivered via filter paper. A subset of 86 patients reported weekly vegetable intakes, excluding salad or potatoes. PROP bitterness correlated significantly with polyp number, an effect separate from age-associated increases in polyp number. The PROP-polyp relationship was strongest in men over 66 years, and older men with polyps were most likely to be overweight or obese. In the subset reporting vegetable intake, men who tasted PROP as more bitter consumed fewer vegetables. These preliminary findings suggest that taste genetics may influence colon cancer risk, possibly through intake of vegetables. PMID- 15810631 TI - Butyrate-induced differentiation of colon cancer cells is PKC and JNK dependent. AB - Butyric acid, a short-chain fatty acid physiologically present in human large gut, is derived from bacterial fermentation of complex carbohydrates. It has been shown to reduce the growth and motility of colon cancer cell lines and to induce cell differentiation and apoptosis. Apoptosis is considered a result of normal colonocyte terminal differentiation in vivo. The aim of this study was to characterize the cellular mechanisms regulating differentiation of colon cancer cells stimulated with sodium butyrate (NaB). The two human colon cancer cell lines Caco-2 and HT-29 were treated with NaB at physiologically relevant concentrations. Alkaline phosphatase (ALP) activity, a marker of colonocyte differentiation, was increased 48 hr after treatment with 1 mM NaB. Higher doses of NaB (5 and 10 mM) induced apoptosis of the cells and failed to stimulate the colonocyte differentiation. Therefore, we assumed that butyrate augments cell differentiation and induces apoptosis, acting via various intracellular mechanisms, and butyrate-mediated programmed cell death cannot be considered a consequence of colonocyte terminal differentiation. The effect of NaB on ALP activity was significantly attenuated in the presence of inhibitors of protein kinase C and JNK. Inhibition of MEK-ERK signal transduction pathways augmented the impact of butyrate on colonocyte differentiation. These results suggest that butyrate could influence the colonocyte differentiation via modulation of the activity of cellular protein kinases and signal transduction. PMID- 15810632 TI - Prospective study of the association of serum triglyceride and glucose with colorectal cancer. AB - To determine if serum triglyceride and glucose levels are associated with colorectal cancer, a prospective study among 7619 Japanese-American men was conducted. From 1968 to 1998, 376 colon and 124 rectal cancer incident cases were diagnosed. A strong positive association of alcohol intake and pack-years of cigarette smoking with colorectal cancer was observed. Body mass index and heart rate were also positively related to colon, but not to rectal cancer. In contrast, serum triglyceride did not predict the development of either colon or rectal cancer. There was a modest association of serum glucose in the highest quartile group with rectal cancer (relative risk = 1.33; 95% confidence interval, 0.79-2.26), but it was not statistically significant. This study did not find a strong positive association of serum triglyceride or glucose with colorectal cancer, but additional studies including other metabolic consequences associated with increased serum triglyceride and glucose may clarify the relationship. PMID- 15810633 TI - Ganglioneuromatous polyposis associated with juvenile polyps and a tubular adenoma. PMID- 15810634 TI - Multifocal gastric carcinoid tumor in a patient with pernicious anemia receiving lansoprazole. PMID- 15810635 TI - A nuclear receptor ligand down-regulates cytosolic phospholipase A2 expression to reduce bile acid-induced cyclooxygenase 2 activity in cholangiocytes: implications of anticarcinogenic action of farnesoid X receptor agonists. AB - Bile acids are considered to be involved in the development of biliary tract carcinoma, although the underlying mechanisms are yet to be established. The aims of this study were (1) to investigate the carcinogenic role of bile acids in the biliary system based on the arachidonate-prostanoid pathway and (2) to clarify the therapeutic role of a farnesoid X receptor (FXR) ligand that modifies bile acid metabolism. Immortalized mouse cholangiocytes were incubated with glycochenodeoxycholate (GCDC), taurocholate, taurochenodeoxycholate, taurodeoxycholate, and tauroursodeoxycholate. GCDC induced cyclooxygenase 2 (COX 2) expression (Western blotting, 1.7-fold; RT-PCR, 2.3-fold) and prostaglandin (PG) production (PGE2, 6.3-fold; PGF2alpha, 8.5-fold), whereas cytosolic phospholipase A2 (cPLA2) expression and activity were reduced. In contrast, no marked changes were induced by the other bile acids. When the same experiment was performed in the presence of a synthetic FXR ligand (GW4064), cPLA2 expression and activity were reduced, although COX-2 expression was unchanged. GW4064 also suppressed PG generation by 40%. In conclusion, the present findings suggest a carcinogenic potential of GCDC. A synthetic FXR ligand (GW4064) inhibited the induction of COX-2 activity (detected as PG production) by GCDC, suggesting its anticarcinogenic potential. This effect seemed to be due to down-regulation of cPLA2. FXR ligands may have therapeutic potential against biliary carcinogenesis, but a delivery system for these agents is still to be developed. PMID- 15810636 TI - Associated factors in modulating aflatoxin B1-albumin adduct level in three Chinese populations. AB - To elucidate the potential factors modulating exposure to aflatoxin B1 (AFB1) in three Chinese populations, an epidemiologic study was conducted in Fusui County and Nanning City of Guangxi Province and Chengdu City of Sichuan Province. The incidence rates of hepatocelluar carcinoma (HCC) for males in these three regions were 92-97 per 100,000, 32-47 per 100,000, and 21 per 100,000, respectively. Eighty-nine residents from Fusui, 196 residents from Nanning, and 118 residents from Chengdu were screened for AFB1-albumin adduct (AAA) levels and hepatitis virus (HBV, HCV, HDV, HEV, and HGV) infections, as well as liver biochemistry (alanine aminotransferase [ALT], aspartate aminotransferase [AST], alkaline phosphatase [ALP], y-glutamyl transpeptidase [GGT], 5'-nucleotidase, globulin [GLO], direct bilirubin, indirect bilirubin, and bile acid levels). At least one marker of hepatitis virus (HV) infection was present in 47.2% (42/89) of subjects from Fusui, while in Nanning and Chengdu the values were 15.8% (31/196) and 22.0% (26/118), respectively. In contrast to females, a higher level of AAA was observed in males; the difference was statistically significant in both the Nanning (P = 0.023) and the Chengdu (P = 0.026) subjects. In the Chengdu group, there was a significantly higher level of AAA in cases with HV infection (P = 0.041). There was a close association between AAA level and BMI in the adults without HV infection (r = 0.148, P = 0.044). Also, AAA was closely associated with DBIL and GGT in non-HV-infected minors (P < 0.05), closely associated with ALB, GLO, and GGT in HV-infected minors (P < 0.05), and closely associated with IBIL, GLO, TBA, and AST in non-HV-infected adults (P < 0.01). The co-effect of HV infection and AFB1 exposure may be responsible for the high risk of HCC in the Fusui region, whereas age, gender, BMI, and HV infection may modify individual aflatoxin levels. The relationship between AAA level and liver biochemistry indicates injury induced by aflatoxin to both hepatic parenchyma and biliary tract. But the associations vary with age and HV infection status. PMID- 15810638 TI - Fulminant liver failure associated with the use of black cohosh. PMID- 15810637 TI - Clinical management of hepatic malignancies: ferucarbotran-enhanced magnetic resonance imaging versus contrast-enhanced spiral computed tomography. AB - This study compares ferucarbotran-enhanced magnetic resonance (MR) imaging with spiral computed tomography (CT) in the clinical management of patients with suspected hepatic malignancy. One hundred fifty-nine patients with suspected liver malignancy were included in a prospective, multicenter, phase IIIB clinical trial. Patients were examined with baseline plus contrast-enhanced spiral CT and with baseline plus ferucarbotran-enhanced MR imaging. For each patient, four maps of liver lesions were constructed by a trusted blind party on the basis of (a) baseline plus contrast-enhanced spiral CT, (b) baseline MR imaging, (c) baseline plus ferucarbotran-enhanced MR imaging, and (d) final diagnosis. Three blinded experts evaluated the treatment options for each map. The concordance of the treatment choice based on imaging with the treatment choice based on final diagnosis was verified. The final therapeutic decision was radical therapy for 85 patients, palliative therapy for 49 patients, and no treatment for 25 patients who had a final diagnosis of benign lesion. Proper treatment allocation was achieved in 121 (76%) of 159 patients when the evaluation was based on liver maps obtained from spiral CT, in 97 (61%) of 159 patients when the evaluation was based on liver maps obtained from baseline MR imaging, and in 136 (85%) of 159 patients when the evaluation was based on liver maps obtained from baseline plus ferucarbotran-enhanced MR imaging. MR imaging with ferucabotran was significantly superior to spiral CT for correct treatment allocation (P = 0.0167). Ferucarbotran-enhanced MR imaging is superior to spiral CT in clinical management of patients with suspected hepatic malignancy. PMID- 15810639 TI - Management of Budd-Chiari syndrome. AB - Thrombotic occlusion of the hepatic veins leads to liver dysfunction and liver failure requiring liver transplantation in advanced cases. The cause for the occlusion of the hepatic veins is not completely understood. However, several underlying conditions such as polycytemia, factor V Leiden mutation, and protein C and S deficiency are found in these patients. We here report our single-center experience with 18 consecutive patients with Budd-Chiari Syndrome (BCS) who were treated at our institution between August 1992 and June 2003. Twelve patients underwent liver transplantation, three patients received stents into the hepatic veins or vena cava, another patient was treated with TIPSS (transjugular intrahepatic postosystemic stent shunt), and one patient underwent surgical mesocaval shunting. Three patients, among those the patient with TIPSS, were put on anticoagulant therapy and are scheduled for liver transplantation. We outline the indication for an approach tailored to the stage of the disease and the adaption of the procedures with the deterioration of clinical conditions. Surgical aspects and postoperative management with a focus on liver transplantation are outlined. We conclude from our observations that the management of BCS requires an approach that exhausts conservative approaches until clinical conditions deteriorate with respect to portal hypertension or liver function. Conservative management, i.e., interventional and supportive medical therapy, has been used up to 8 years in our series, until the time for liver transplantation is reached. Liver transplantation for BCS had more complications than transplantation for other liver diseases in our series. Therefore, we propose to keep liver function stable using interventional techniques to maintain venous outflow. If venous outflow cannot be interventionally restored and liver function deteriorates or cirrhosis develops during this time course, liver transplantation is the therapy of choice. PMID- 15810640 TI - Impact of immunosuppressive treatment on liver fibrosis in autoimmune hepatitis. AB - The impact of treatment on progression of fibrosis in autoimmune hepatitis (AIH) is unknown. We assessed the changes in liver fibrosis before and after treatment among these patients. Nineteen AIH patients who had paired liver biopsies were studied. Of these, seven had been treated with 6 months of cyclosporine A and the rest with 6 months of prednisolone for induction of remission. Thereafter all had been maintained on azathioprine. Biopsy specimens before and after treatment were reviewed by one pathologist and scored by the Ishak method. Mean fibrosis stages before and after treatment were compared. Also, factors predicting significant fibrosis (stage > or =3) and cirrhosis (stage > or =5) at presentation were assessed. Mean interval between biopsies was 3.38 years. Mean fibrosis stage decreased from 4.53 to 2.16 following treatment (P < 0.001). Mean decrement in inflammatory grade was 8 scores (range, 4-10) in patients in whom fibrosis improved, and 2 scores (range, 0-4) in patients in whom fibrosis did not decrease after treatment (P < 0.001). ALT-to-platelet ratio was the best predictor of significant fibrosis and also cirrhosis. Fibrosis commonly improves after immunosuppressive treatment in AIH. ALT-to-platelet ratio can predict accurately the presence of significant fibrosis and cirrhosis in AIH. PMID- 15810641 TI - Cholangiohepatoma presenting with recurrent cholangitis. PMID- 15810642 TI - Uncoupling protein 2 involved in protection of glucagon-like peptide 2 in small intestine with ischemia-reperfusion injury in mice. AB - Glucagon-like peptide 2 (GLP-2) is an intestinal epithelium-specific growth factor. However, its protective effects and related mechanism on the small intestine injured by ischemia-reperfusion (I/R) in mice remain unclear. This study aimed to reveal the effects of GLP-2 and its functional relationship with uncoupling protein 2 (UCP2) on the small intestine after I/R injury in mice. Male Balb/c mice were given GLP-2 (250 microg/kg/day, ip) for 3 days and underwent 30 min of superior mesenteric artery occlusion followed by 1 hr of reperfusion on day 4. Histological damage, bacterial translocation, diamine oxidase, and malondialdehyde level were assessed, and UCP2 expression was measured by immunohistochemistry and Western blot. GLP-2 attenuated the intestinal histological damage caused by I/R and increased the villous height by 28% and the crypt depth by 10%, respectively. Compared to the I/R group, diamine oxidase activity was increased, the incidence of bacterial translocation and malondialdehyde level were decreased, and UCP2 expression was increased in GLP-2 treated mice. GLP-2 protected the small intestine from I/R injury and increased UCP2 expression. These results suggested that effects of GLP-2 should be related to the upregulation of mitochondrial UCP2, which antagonized reactive oxygen species production. PMID- 15810643 TI - Multiple myeloma presenting as mesenteric venous thrombosis and intestinal infarction. PMID- 15810644 TI - Inhibition of Clostridium difficile toxin A-induced colitis in rats by APAZA. AB - A new compound, APAZA, consisting of a molecule of 5-aminosalicylic acid linked to one molecule of 4-aminophenylacetic acid by an azo bond, was testedfor its ability to inhibit acute colitis in rats caused by Clostridium difficile toxin A. When administered chronically for 5 days in drinking water, APAZA significantly inhibited toxin A-induced myeloperoxidase activity, luminal fluid accumulation, and structural damage to the colon at doses of from 1 to 100 mg/kg x day. For comparison, sulfasalazine was administered in identical doses and was found to significantly inhibit toxin A-induced colitis only at the dose of 100 mg/kg x day. When 4-aminophenylacetic acid alone was administered chronically in drinking water, it also inhibited toxin A-induced colonic inflammation at a dose of 100 mg/kg x day. In order to determine if 4-aminophenylacetic acid has a direct anti inflammatory effect on the colon rather than a systemic effect, 4 aminophenylacetic acid was administered acutely to surgically prepared isolated colonic segments by intraluminal injection in anesthetized rats 30 min before toxin A was injected. 4-Aminophenylacetic acid strongly and significantly inhibited toxin A-induced colitis in this experiment at doses as low as 10 microg/segment. It is concluded that APAZA is a potent inhibitor of toxin A induced colonic inflammation in rats and that its constituent, 4 aminophenylacetic acid, is responsible for this increased protection against colitis compared to the 5-aminosalicylic acid component of sulfasalazine. PMID- 15810645 TI - Circulating cell-derived microparticles in Crohn's disease. AB - Procoagulant membrane microparticles can be released from activated or apoptotic cells in response to various environmental stimuli. The aim of this study was to investigate the presence of microparticles in Crohn's disease and to assess their variations after infliximab therapy. We compared the levels of circulating microparticles in 38 patients with Crohn's disease, 16 patients with ulcerative colitis, 7 patients with infectious colitis, and 17 control subjects. The evolution of microparticle levels was assessed after infliximab therapy in 13 patients with Crohn's disease. Circulating microparticle levels were elevated in patients with Crohn's disease (9.31+/-0.66 nmol/L phosphatidylserine equivalent [PS Eq]) or infectious colitis (10.71+/-0.92 nmol/L PS Eq) compared to patients with ulcerative colitis (5.75+/-0.59 nmol/L PS Eq) and control subjects (4.06+/ 0.37 nmol/L PS Eq) (P = 0.001). Infliximab induced a significant diminution of the amounts of circulating microparticles, from 10.33+/-1.20 to 6.45+/-0.90 nmol/L PS Eq (P = 0.002). Generation of circulating microparticles occurs in Crohn's disease; infliximab induces significant diminution. Release of microparticles could be linked to the type of inflammatory response underlying Crohn's disease. PMID- 15810646 TI - Efficacy of mesalazine in the treatment of symptomatic diverticular disease. AB - We aimed to improve symptoms by means of mesalazine in symptomatic colonic diverticular disease patients. One hundred seventy outpatients (98 M, 72 F; age, 67.1 years; range, 39-84 years) were assigned to four different schedules: rifaximin, 200 mg bid (Group R1: 39 pts), rifaximin, 400 mg bid (Group R2: 43 pts), mesalazine, 400 mg bid (Group M1: 40 pts), and mesalazine, 800 mg bid (Group M2: 48 pts), for 10 days per month. At baseline and after 3 months we recorded 11 clinical variables (upper/lower abdominal pain/discomfort, bloating, tenesmus, diarrhea, abdominal tenderness, fever, general illness, nausea, emesis, dysuria), scored from 0 = no symptoms to 3 = severe. The global symptomatic score was the sum of all symptom scores. After 3 months in all schedules but Group R1, 3 of the 11 symptoms improved (P < 0.03); the global score decreased in all groups but Group R1 (P < 0.0001). Mesalazine-treated patients had the lowest global score at 3 months (P < 0.001). Mesalazine is as effective as rifaximin (higher dosage schedule) for diminishing some symptoms, but it appears to be better than rifaximin for improving the global score in those patients. PMID- 15810647 TI - Case-finding for adult celiac disease in patients with reduced bone mineral density. AB - The aim of this study was to assess the value of case-finding for unrecognized adult celiac disease (CD) in patients with reduced bone mineral density (BMD), verified by dual-energy X-ray absorptiometry (DXA). Patients attending for a DXA scan were investigated for CD using immunoglobulins, IgG/IgA antigliadin antibodies (AGA), and endomysial antibodies (EMA). All patients with a positive IgA AGA, EMA, or only IgG AGA in the presence of IgA deficiency had a small bowel biopsy. There were 12 cases of CD (12/978), a prevalence of 1.2% for the whole cohort. The prevalence of CD was 0.7% (2/304) for those with a normal BMD, 1.2% (5/431) for patients with osteopenia, and 2.1% (5/243) for patients with osteoporosis. Direct questioning revealed that all patients with unrecognized CD had subtle gastrointestinal symptoms or a history of anemia. Excluding patients without these symptoms would give a prevalence of 3.9% for osteoporosis (5/127) and 2.6% for osteopenia (5/191 ). This study suggests that there is no value of unselected case-finding for CD in patients with a reduced BMD. However, a targeted case-finding approach may be more valid and cost-effective with the initial selection of patients who should be investigated for CD based on questioning about gastrointestinal symptoms or anemia. PMID- 15810649 TI - High-resolution magnifying upper endoscopy in a patient with patchy celiac disease. PMID- 15810648 TI - Cytokine production by intestinal intraepithelial lymphocyte subsets in celiac disease. AB - One of the earliest signs of mucosal immune activation in celiac disease (CD) is an increase in the intraepithelial lymphocyte (IEL) count in the small intestinal epithelium. Though most of those IELs express T cell receptor (TcR)-alphabeta chains, CD is characterized by an increase in TcR-gammadelta+ IELs and by the loss of CD3- IELs. There is currently little evidence that these changes in IEL subset distribution are of relevance in the pathogenesis of CD. We aimed to determine the pattern of cytokine production by IEL subsets isolated from duodenal biopsy specimens from control subjects and CD patients at different stages of the disease. We quantified the capacity of IEL subsets to produce IFN gamma, TNF-alpha, IL-2, IL-4, and IL-10 by intracellular staining by flow cytometry. All IEL subsets studied displayed a type I cytokine profile in both CD and control subjects, with TcR-alphabeta+ IELs being the main IFN-y producers. Untreated CD exhibited a trend toward a superior accumulation of IFN-gamma per cell but a reduced proportion of INF-gamma+ cells in vitro in association with a significantly increased apoptotic rate of IELs. IL-4 was almost undetectable in all cases and IL-10 showed a tendency to increase in treated and "silent" celiac patients. IEL subsets have a similar Th1 profile in controls and CD patients, and the superior in vitro apoptosis of IELs from CD patients may reflect their superior in vivo activation. The induction of IL-10-dependent regulatory Tr1 responses may be of potential clinical significance in this disease and merits further investigation. PMID- 15810650 TI - Is there a relation between irritable Bowel syndrome and urinary stone disease? AB - Our aim was to investigate the role of renal colic, a clinical condition characterized by excruciating pain, in the etiopathogenesis of irritable bowel syndrome (IBS). Two groups of patients were enrolled in the study. Group I consisted of 59 patients (33 male and 26 female) with a median age of 41.9 (18 to 58) years. The patients in group I were admitted to our clinic with urinary stone disease and with a medical history of acute renal colic. Group II consisted of 55 patients (25 male and 30 female) with a median age of 40.1 (18 to 56) years, complaining of urologic abnormalities other than stone disease. IBS was diagnosed using Rome criteria. Metabolic analysis for stone disease was performed on patients in group I. The incidence of five metabolic abnormalities--low urine volume, hypercalciuria, hyperoxaluria, hyperuricosuria and hypocitraturia--in patients with and without irritable bowel disease was investigated. IBS was found in 16 of the 59 patients (27.1%) in group I and in 6 of the 55 patients (10.9%) in group II. The difference was statistically significant (P < 0.05). Relative risk of developing IBS was 2.48 times higher in patients with urinary stone disease than in those without stone disease. There was no statistically significant difference in the metabolic analysis of patients with and without IBS in group I. IBS causes great suffering. Urinary stone disease should be considered as an etiological factor during management of IBS patients. In the presence of gastrointestinal symptoms, a patient with a medical history of acute renal colic might be referred to a gastroenterologist. PMID- 15810651 TI - Optimising plasticity: environmental and training associated factors in transplant-mediated brain repair. AB - With progressively ageing populations, degeneration of nerve cells of the brain, due to accident or disease, represents one of the major problems for health and welfare in the developed world. The molecular environment in the adult brain promotes stability limiting its ability to regenerate or to repair itself following injury. Cell transplantation aims to repair the nervous system by introducing new cells that can replace the function of the compromised or lost cells. Alternatives to primary embryonic tissue are actively being sought but this is at present the only source that has been shown reliably to survive grafting into the adult brain and spinal cord, connect with the host nervous system, and influence behaviour. Based on animal studies, several clinical trials have now shown that embryonic tissue grafts can partially alleviate symptoms in Parkinson's disease, and related strategies are under evaluation for Huntington's disease, spinal cord injury, stroke and other CNS disorders. The adult brain is at its most plastic in the period following injury, offering a window of opportunity for therapeutic intervention. Enriched environment, behavioural experience and grafting can each separately influence neuronal plasticity and recovery of function after brain damage, but the extent to which these factors interact is at present unknown. To improve the outcome following brain damage, transplantation must make use of the endogenous potential for plasticity of both the host and the graft and optimise the external circumstances associated with graft-mediated recovery. Our understanding of mechanisms of brain plasticity subsequent to brain damage needs to be associated with what we know about enhancing intrinsic recovery processes in order to improve neurobiological and surgical strategies for repair at the clinical level. With the proof of principle beginning to emerge from clinical trials, a rich area for innovative research with profound therapeutic application, even broader than the specific context of transplantation, is now opening for investigation. PMID- 15810652 TI - Function of cGMP-dependent protein kinases in the nervous system. AB - The second messenger cyclic guanosine-3',5'-monophosphate (cGMP) mediates many effects of nitric oxide in the nervous system. cGMP may act through various intracellular receptors, among them a family of serine/threonine kinases, the cGMP-dependent protein kinases (cGKs). Hitherto, three mammalian cGKs have been identified: cGKIalpha, cGKIbeta and cGKII. Discrete functions of cGKI and cGKII are determined by their distinct expression patterns and targeting to specific substrates. This review provides an overview about the expression and functions of cGKs in the nervous system. Main emphasis is put on the discussion of phenotypes observed in cGK-deficient mouse models that lack cGKI and/or cGKII globally or selectively in brain regions of interest. Recent data demonstrate important functions of cGKI in (1) the development and sensitization of nociceptive neurons, and (2) synaptic plasticity and learning. There is also evidence suggesting that cGKII in the suprachiasmatic nucleus of the hypothalamus is involved in the regulation of circadian rhythmicity. Thus, cGKs serve key functions in the transduction of cGMP signals into cellular responses in distinct regions of the nervous system.- PMID- 15810653 TI - Chronic stress, depression and antidepressants: effects on gene transcription in the hippocampus. AB - Depressive disorders are among the most frequent forms of mental illness. Both genetic and environmental factors, such as stress, are involved in the etiology of depression. Therefore, chronic stress paradigms in laboratory animals constitute an important tool for research in this field. The molecular bases of chronic stress/depression are largely unknown, although a large amount of information has been accumulated during recent years. Dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis as well as structural and physiological alterations in the hippocampus and neocortex are known to occur. Modifications in the expression level of some genes, such as brain-derived neurotrophic factor, cAMP-response-element binding protein, serotonin receptors and HPA axis components were consistently associated in a number of experimental models. However, recent results suggest that several synaptic proteins, transcription factors and proteins involved in neuronal growth/differentiation, are also modified in their expression in experimental models of chronic stress. In general, these alterations can be reversed by treatment with antidepressants. Thus, a complex pattern of gene expression leading to stress/depression is starting to emerge. We summarize here recent findings on the alterations of gene expression in the hippocampus of chronically stressed and antidepressant treated animals. PMID- 15810654 TI - Targeting the JNK pathway as a therapeutic protective strategy for nervous system diseases. AB - The c-Jun N-terminal kinases (JNKs) are members of the family of mitogen activated protein kinases (MAPKs). While the functions of the JNKs under physiological conditions are diverse and not completely understood, there is increasing evidence that JNKs are potent effectors of apoptosis in both the brain and the mammalian inner ear following a variety of injuries. The activation of the inducible transcription factor c-Jun by N-terminal phosphorylation is a central event in JNK-mediated neural and inner ear hair cell death. A cell permeable peptide designed specifically to inhibit JNK signaling has proven successful in in vivo models of both neuronal degeneration following cerebral ischemia and auditory hair cell degeneration following exposure to either acoustic trauma or a toxic level of an aminoglycoside antibiotic. Here we discuss the evidence supporting the application of JNK inhibitors to prevent cellular degeneration in several central nervous system (CNS) and peripheral nervous system (PNS) diseases with an emphasis on traumatic ischemic damage to the CNS and acquired deafness in the PNS receptors. PMID- 15810655 TI - The role of neural synchronization in the emergence of cognition across the wake sleep cycle. AB - Searching for the neural code underlying consciousness and cognition is one of the most important activities in contemporary neuroscience. Research with neuronal oscillations at the level of single-neuron, local cell assemblies, and network system have provided invaluable insights into different mechanisms of synaptic interactions involved in the emergence of cognitive acts. A cognitive neuroscience of conscious experience is gradually emerging from behavioral and neuroimaging studies, which can be successfully complemented with the quantitative EEG findings discussed here. This review is an attempt to highlight the value of state-dependent changes in human neurophysiology for a better understanding of the neurobiological substrate underlying those aspects of cognition drastically affected by sleep states. Recent advances related to synchronization mechanisms potentially involved in brain integration processes are discussed, emphasizing the value of scalp and intracranial EEG recordings at determining local and large-scale dynamics in the human brain. Evidence supporting the critical role of state-dependent synchrony in brain integration comes mainly from studies on the theta and gamma oscillations across the wake sleep continuum, as revealed by human intracranial recordings. This review blends results from different levels of analysis with the firm conviction that state dependent brain dynamics at different levels of neural integration can provide a deeper understanding of neurobiological correlates of consciousness and sleep functions. PMID- 15810656 TI - [Morphogenes of Hydra sp]. PMID- 15810657 TI - [Evolution of mechanisms of Calcium signaling: the role of Calcium ions in signal transduction in prokaryotes]. PMID- 15810658 TI - [Antioxidants activity in intestines of larvae Galleria mellonella L. (Lepidoptera, Pyralidae) during Bacillus thuringiensis ssp Galleriae infection]. PMID- 15810659 TI - [Humoral reactions of starfish Asterias rubens innate immunity]. PMID- 15810660 TI - [Comparative characteristics of absorbtion spectra of hemoglobins of different species of fish]. PMID- 15810661 TI - [Na,K-atpase activity in different brain regions in cerebral ischemia and influence of quinacrine and indomethacin administration]. PMID- 15810662 TI - [Topography of digestive enzymes in small intestine and activity of hydrolases in liver and kidneys of adult pigs]. PMID- 15810663 TI - [Chemical mechanisms of regulation of muscle contraction in Lymnaea stagnalis L]. PMID- 15810664 TI - [Fucoidans influence on developing embryos of sea urchin Strongylocentrotus intermedius]. PMID- 15810665 TI - [Morphofunctional characteristics oh lamprey Lethenteron japonicum heart]. PMID- 15810666 TI - [Dove Columba livia thermoregulation during stress induced by food deprivation]. PMID- 15810667 TI - [The role of sympathetic and parasympathetic mechanisms in formation of heart secondary rhythms during rat ontogenesis]. PMID- 15810669 TI - [Effects of sleep deprivation on rats with congenital susceptibility to audiogenic seizures]. PMID- 15810668 TI - [Nociceptive sensitivity to longterm exposure to an irritant in formaline test of males and females of rat in postnatal development]. PMID- 15810670 TI - [Comparative morphological analysis of spleen of perch Perca Fluviatilis from lakes with different water pH]. PMID- 15810671 TI - [Histochemical analysis of influence of weak electromagnetic field on structures of rat mesencephalon]. PMID- 15810672 TI - [A concept of relation between physical and biological time in animals]. AB - A definition is proposed for biological ("internal") time tau(t) for a growing organism whose weight variation obeys the law w(t): tau(t) = 1/c(w) (t) = w(t)/w'(t), where t is physical ("external") time, w'(t) is weight increase rate, and c(w) (t) = w'(t)/w(t) is specific growth rate. Properties of functions tau(t) and w(tau) were studied for those cases when growth curves w(t) were described by Bertalanffy's or logistic equations. PMID- 15810673 TI - [Molecular mechanisms of macroevolution]. AB - The present paper is devoted to the evolutionary role of genetic modules shuffing. The mechanisms capable to produce new molecular functions and significant complications of ontogenesis are reviewed. Two-step model of macroevolution is proposed. This model comprises: (1) Arising of a new combination of genetic modules. This step does not result in formation of a new taxon but makes necessary ground for that process. (2) Precise structure completing of the new combination of modules and corresponding genome optimization by use of various mechanisms including point mutations. This step concerns many genes and finally leads to formation of a new taxon. It is shown that arising of new combinations of genetic modules might work out as molecular basis for progressive evolution, while alternative structural completing of the same combination might result in adaptive radiation. PMID- 15810674 TI - [Continuity and discontinuity of the geomerida: the bionomic and biotic aspects]. AB - The view of the spatial structure of the geomerida (Earth's life cover) as a continuum that prevails in modern phytocoenology is mostly determined by a physiognomic (landscape-bionomic) discrimination of vegetation components. In this connection, geography of life forms appears as subject of the landscapebionomic biogeography. In zoocoenology there is a tendency of synthesis of alternative concepts based on the assumption that there are no absolute continuum and absolute discontinuum in the organic nature. The problem of continuum and discontinuum of living cover being problem of scale aries from fractal structure of geomerida. This problem arises from fractal nature of the spatial structure of geomerida. The continuum mainly belongs to regularities of topological order. At regional and subregional scale the continuum of biochores is rather rare. The objective evidences of relative discontinuity of the living cover are determined by significant alterations of species diversity at the regional, subregional and even topological scale Alternatively to conventionally discriminated units in physionomically continuous vegetation, the same biotic complexes, represented as operational units of biogeographical and biocenological zoning, are distinguished repeatedly and independently by different researchers. An area occupied by certain flora (fauna, biota) could be considered as elementary unit of biotic diversity (elementary biotic complex). PMID- 15810675 TI - [Seasonal succession of lotic epiphyton: effects of macrophyte cover and grazing by chironomid larvae]. AB - The epiphyton is a community of periphyton on the aquatic plants. The seasonal dynamics of biomass, production and community structure of epiphytic algae on Potamogeton perfoliatus was studied at the Moscow River reach with low flow velocity from May to October 2000. The relative importance of physical-chemical environmental variables and herbivorous pressure in structuring the epiphyton community was accessed using multivariate redundancy analysis. Algal composition was mainly determined by the macrophyte cover, suggestion the overriding importance of the underwater light availability in seasonal development of epiphyton. There was a significant positive relationship between biomass of attached diatom Cocconeis placentula and abundance of chironomid larvae (r = 0.57; p < 0.01). This indicates that increase in herbivorous pressure caused the shift in epiphyton community structure. However, the intensity of herbivorous impact on epiphyton was ultimately determined by light availability in macrophyte stands. At low irradiance level and consequent slow growth of algae the upper layer of epiphyton was seriously destroyed by chironomids that resulted in dominance of C. placentula. As irradiance level and epiphyton productivity increased chironomids were unable to prevent biomass accumulation of the upper layer thus inhibiting the growth of C. placentula. The high concentrations of nutrients and relatively stable discharge determined the crucial role of light regime in the regulation of trophic interactions between epiphyton and chironomid larvae. PMID- 15810676 TI - [Change in the system of wing venation of Drosophila melanogaster under heat shock and selection]. AB - A change in the system of wing venation of Drosophila melanogaster appeared in response to heat shock and positive selection pressure directed to restoring the normal formation of wing radial vein, L2, that had been violated earlier by the recessive mutation of radius incompletes. Positive selection was effective, L2 having been formed correctly and completely to 35 generation. Besides, (+) selection was accompanied by appearance of a small fragment of an additional vein at the wing tip. Selection directed to increase of size of this fragment resulted in the sufficient changes in the system of wing venation as a whole. It is suggested that, during evolution, transformation of wing venation of Drosophila was effected by the change of the way of prepattern realization, whereas the cells of wing plate continued to allow the formation of veins practically over a whole area. PMID- 15810677 TI - [Retransmission of echo-like signals: methods and results of studies on nocturnal moths (Insecta)]. AB - Studies of capability to echolocate in nocturnal moths using the method of echo signal retransmission are described in detail. During retransmission the insect was presented not with the echo from some real object but with electronically generated echo-like signal which appears following the certain delay after insect's own acoustic signal. In that way the artificial echo from non-existent obstacle is produced. In the current study the delay was set to 0.5 ms that corresponds to a distance of about 8 cm from obstacle. At the same time there was no any single movement of any object near the flying insect therefore the activities of sensory systems other than auditory did not cause systematical influence on results obtained. Noctuid moths (Noctuidae, Lepidoptera) are acoustically active insects. They are capable of producing ultrasonic clicks during flight. The most typical response of a moth to the retransmission of echo like stimuli is an activation of own clicks emission that is often accompanied with rise in click amplitude. Using the activation of emission as a criterion of moth's response to the echo-like stimulation we measured echolocational thresholds in three species: Amphipyra pyramidea (36 dB SPL), Enargia paleacea (31 dB SPL) and Blepharita satura (26 dB SPL). The ability to echolocate was also demonstrated in 20 species of subfamilies Catocalinae, Amphipyrinae, Cuculliinae, Hadeninae, Noctuinae, Heliothinae. PMID- 15810678 TI - [On methodological aspects of ecological experiments (comments on M. V. Kozlov publication)]. AB - The meaning of conception of pseudoreplication in ecological experiments is discussed. The replications are found to be pseudo if factors causing random variation act on such replications as on one unit. For example, preudoreplications could appear if a single flipped coin would be split up. The absence of random spatial distribution of experimental and control fields could be considered as a mistake because the experimental results could interfere with spatial heterogeneity. However, there are no pseudoreplications in this case, because the variations of ecological systems are produced by factors that are different in different places and each organism responds to these factors individually. The pseudoreplication should be created by multiple determination of a response of an individual organism. Also recommendations for applications of randomized block design and analysis of covariance for field trials in plant communities are proposed. Randomized blocks unite the experimental and control fields occupying the identical place in the series of spatial heterogeneity. The influence of such heterogeneity on random variation of determined parameters could be excluded by this approach. The heterogeneity considered only at the scale of used blocks. As far as control and experimental fields belongs to a single set, the series of heterogeneities, that comprises the range of their parameters, is identical. The random sorting of the large amount of experimental and control samples consisting of the same number of replications will produces replications in the same place of the series of general heterogeneity of investigated set. The replications occupying the same places of the sorted series form "virtual block". The population of such blocks includes the majority of random variation and allows one to reveal even very weak experimental effects by variance analysis. PMID- 15810679 TI - [Tendencies in auditory physiology]. AB - Main tendencies in studying of human and animals auditory system with psychoacoustical and electrophysiologycal methods are considered. Concerning psychoacoustical studies some basic data are presented as well as contemporary tendencies in hearing physiology in analysis of the intensity, frequency, temporal characteristics of the sound signals and data related to such phenomena as masking and adaptation. Data concerning directional hearing are presented in detail as a basis of auditory virtual reality. In electrophysiological studies of the auditory system detailed analysis of mapping in auditory centers and mechanisms concerning localization of unmoved and moving auditory stimuli was performed. Special attempt was paid to consider the reflection of different types of auditory signals in human evoked potentials. PMID- 15810680 TI - [Evolutionary theories of asymmetrization of organisms, brain and body]. AB - Growth of a dispersion of elements of unitary systems (US) inevitably transfrorms them into the binary connected differentiations (BCD). So, at a level of genes, from bisexuals have arisen females and males, and at a level of hormones (mentality, behavior), from symmetric--asymmetric: functions, organs, right handers and left-handers. All BCD are isomorphic. They consist of subsystems preservation (conservative) and change (operative) (SP, SC). SP is more important, than SC, therefore the dispersion of their elements is less, than elements SC. This base difference. It transforms a monomodal population in bimodal, direct "ecology, (environment, E US), in consequetn (E SC SP), and synchronous evolution in asynchronous (at first SC, later SP). Then for evolution "pay" only SC. Means, asynchronous evolution is more economical, than synchronous. In it adaptive sense of any BCD. All theories of biology are theories of US. They treat subsystems not as phased but as forms, therefore may not explain BCD. The general idea of asynchronous evolution--control centre of any function arises in the left hemisphere and in the right hemisphere gets only after approbation. Hence, creates asymmetry different phases of the same function in the left and right hemisphere, but not different functions. It has allowed to reject a lot of erroneous representations and to create adaptive, internally consistent, evolutionary theories of three-demensional asymmetry of organisms, a brain, a cis-trans asymmetry of pair organs and dextrality-sinistrality, having unique explanatory and predictive potential. PMID- 15810681 TI - [Neuroendocrine effect of sex hormones]. AB - The paper provides a generalization of data and the results of own experiments on influence ovarian steroids on the hypothalamus and other brain areas related to reproduction. Ovarian hormones have widespread effects throughout the brain: on catecholaminergic neurons and serotonergic pathways and the basal forebrain cholinergic system, as well as the hipocampus, spinal cord, nigrostriatal and mesolimbic system, in addition to glial cells and blood-brain barrier. The widespread influences of these various neuronal systems ovarian steroids have measurable effects on mood and affect as well as on cognition, with implications for dementia. There are developmentally programmed sex differenced in hippocampal structure that may help to explain differences in the strategies which male and female rats use to solve spatial navigation problems. The multiple sites and mechanisms of estrogen action in brain underlie a variety of importants effects on cognitive and other brain functions--coordination of movement, pain, affective state, as well as possible protection in Alzheimer's disease. Estrogen withdrawal after natural or surgical menopause can lead to a host of changes in brain function and behavior. PMID- 15810682 TI - [Status epilepticus--the new mechanisms and lines of inhibition (the lithium pilocarpine model)]. AB - This review focuses on the modeling of status epilepticus in animal brain and modern data on the mechanisms of epileptical seizures initiation using the pilocarpine binding with the muscarinic cholinoreceptors (litium pilocarpine model). The character of epileptics seizures in developing brain and adult brain of rats were investigated. The lines of modulation and inhibition epileptics statues by sacricine and intranasal application of neuropeptide thyroliberin in ultra-low doses are demonstrated. The role of the short-term changes (signal regulated kinase signaling cascade, Kv 4.2 potassium channels, hippocampal and cortical spike-wave discharges) and the long-term changes (loss of selective type of interneurons, excitatory circuits by mossy fiber sprouting) that promotion the epileptic state and recurrent seizures in limbic structure are discussed. PMID- 15810684 TI - [Altered states of consciousness]. AB - The review of modern ideas concerning the altered states of consciousness is presented in this article. Various methods of entry into the altered states of consciousness are looked over. It is shown that the altered states of consciousness are insufficiently known, but important aspects of human being existence. The role of investigation of the altered states of consciousness for the creation of integrative scientific conception base is discussed. PMID- 15810683 TI - [Physiological problems of biological evolution. "Errors" in the development of physiological functions]. AB - During the evolution the physiological functions grow more perfect and complex. However, sometimes a developing function can show some evidence of regress and in certain cases can become the cause of dangerous diseases. The autoimmune diseases are among them. We conventionally call such cases the "error" of the evolution. In this work a negative role of leukocytes in the brain microcirculation is considered. With the help of experimental studies it was shown that leukocytes owing to their large volume (by a factor of 2-2.5 greater than the volume of an erythrocyte) slow down the capillary blood flow. As the result of the increasing tendency to adhesion, under the influence of hypoxia they make leukocyte conglomerates which cause the occlusion of venules and smallest veins of the brain. This is the reason for the known effect "no reflow" and results in an organism death. PMID- 15810685 TI - North Pacific Acoustic Laboratory. AB - A series of long-range acoustic propagation experiments have been conducted in the North Pacific Ocean during the last 15 years using various combinations of low-frequency, wide-bandwidth transmitters and horizontal and vertical line array receivers, including a 2-dimensional array with a maximum vertical aperture of 1400 m and a horizontal aperture of 3600 m. These measurements were undertaken to further our understanding of the physics of low-frequency, broadband propagation and the effects of environmental variability on signal stability and coherence. In this volume some of the results are presented. In the present paper the central issues these experiments have addressed are briefly summarized. PMID- 15810696 TI - Statistics and vertical directionality of low-frequency ambient noise at the North Pacific Acoustic Laboratory site. AB - We examine statistical and directional properties of the ambient noise in the 10 100 Hz frequency band from the NPAL array. Marginal probability densities are estimated as well as mean square levels, skewness and kurtoses in third octave bands. The kurotoses are markedly different from Gaussian except when only distant shipping is present. Extremal levels reached approximately 150 dB re 1 micro Pa, suggesting levels 60dB greater than the mean ambient were common in the NPAL data sets. Generally, these were passing ships. We select four examples: i) quiescent noise, ii) nearby shipping, iii) whale vocalizations and iv) a micro earthquake for the vertical directional properties of the NPAL noise since they are representative of the phenomena encountered. We find there is modest broadband coherence for most of these cases in their occupancy band across the NPAL aperture. Narrowband coherence analysis from VLA to VLA was not successful due to ambiguities. Examples of localizing sources based upon this coherence are included. kw diagrams allow us to use data above the vertical aliasing frequency. Ducted propagation for both the quiescent and micro earthquake (T phase) are identified and the arrival angles of nearby shipping and whale vocalizations. MFP localizations were modestly successful for nearby sources, but long range ones could not be identified, most likely because of signal mismatch in the MFP replica. PMID- 15810697 TI - Assessing responses of humpback whales to North Pacific Acoustic Laboratory (NPAL) transmissions: results of 2001--2003 aerial surveys north of Kauai. AB - Eight aerial surveys were flown north of the Hawaiian island of Kauai during 2001 when the North Pacific Acoustic Laboratory (NPAL) source was not transmitting, and during 2002 and 2003 when it was. All surveys were performed during the period of peak residency of humpback whales (Feb-Mar). During 2002 and 2003, surveys commenced immediately upon cessation of a 24-h cycle of transmissions. Numbers and distribution of whales observed within 40 km of the NPAL source during 2001 (source off) were compared with those observed during 2002 and 2003 (source on). A total of 75 sightings was noted during 2001, as compared with 81 and 55 during 2002 and 2003, respectively. Differences in sighting rates (sightings/km) across years were not statistically significant. Assessment of distributional changes relied upon comparisons of three measures: (a) location depths; (b) distance from the NPAL source; and (c) distance offshore. None of the distributional comparisons revealed statistically significant differences across years. Several possible interpretations are examined: (a) whales have habituated to the NPAL signal; (b) insufficient statistical power exists in the present design to detect any effects; and (c) the effects are short-lived and become undetectable shortly after the cessation of transmissions. PMID- 15810698 TI - [At the beginning of an anniversary volume]. PMID- 15810699 TI - [Thoma Ionescu]. PMID- 15810700 TI - Liver transplantation in eastern countries. PMID- 15810701 TI - [Orthotopic liver transplantation in adult patients with cadaveric grafts. Experience of the Fundeni Center of General Surgery and Liver Transplantation]. AB - We analyze the experience of the Center of General Surgery and Liver Transplantation from the Fundeni Clinical Institute (Bucharest, Romania) regarding orthotopic liver transplantation (OLT) in adult recipients, with whole liver grafts from cadaveric donors, between April 2000 (when the first successful LT was performed in Romania) and December 2004. This series includes 37 OLTs in adult recipients (16 women and 21 men, aged between 29-57 years--average 46 years). Other two LT with whole liver cadaveric grafts and two reduced-size LT were performed in children; also, in the same period, due to the acute organ shortage, other methods of LT were performed in 28 patients (21 living donor LT, 6 split LT and one "do mino" LT), that were not included in the present series. The indications for OLT were HBV cirrhosis--10, HBV+HDV cirrhosis--4, HCV cirrhosis--11, HBV+HCV cirrhosis--2, biliary cirrhosis--5, Wilson disease--2, alcoholic cirrhosis--1, non-alcoholic liver disease--1, autoimmune cirrhosis--1. With three exceptions, in which the classical transplantation technique was used, the liver was grafted following the technique described by Belghiti. Local postoperative complications occurred in 15 patients (41%) and general complications in 17 (46%); late complications were registered in 18 patients (49%) and recurrence of the initial disease in 6 patients (16%). Intrao- and postoperative mortality was 8% (3/37). There were two patients (5%) who died because of immunosuppressive drug neurotoxicity at more than 30 days following LT. Four patients (11%) died lately because of PTLD, liver venoocclusive disease, recurrent autoimmune hepatitis and liver venoocclusive disease, myocardial infarction, respectively. Thirty-four patients survived the postoperative period (92%); according to Kaplan-Meier analysis, actuarial patient-survival rate at month 31 was 75%. PMID- 15810702 TI - [Primary retroperitoneal tumors--diagnostic and therapeutic difficulties]. AB - This paper emphasizes the diagnosis and therapeutic difficulties in primary retroperitoneal tumors. There were analyzed 68 primary retroperitoneal tumors (1992-2002): 16 (23.5%) benign tumors, 39 (57.35%) malignant tumors and 13 tumors with unknown histological structure. The preoperative diagnosis was clinically suggested and confirmed by ultrasound and CT examination; the operability was always established by laparotomy. All cases were operated on: complete resection of tumor was possible in 39 cases (57.35%); partial resection in 11 cases (16.17%) and 18 (26.47%) cases were inoperable. There were 5 major intraoperative vascular lesions: 1 inferior vena cava lesion, 1 superior mesenteric vein lesion, 1 left common iliac vein lesion and 2 lumbar artery lesions. Postoperative mortality was represented by 2 cases. Postoperative complication was represented by 3 postoperative hemorrhages, 2 severe pulmonary infections, 1 postoperative evisceration, 1 postoperative acute pancreatitis and 1 acute myocardial infarction. In conclusion the primary retroperitoneal tumors represent a challenge for all surgeons, especially due to surgical approach difficulties, because of problems in tumors intraoperative exploration and resection and because of difficulties in hemostasis. PMID- 15810703 TI - [Minimally invasive approach in gallstone disease complicated with acute pancreatitis]. AB - We have evaluated the minimally invasive approaches (laparoscopy, endoscopy) in the management of the gallstone disease complicated with acute pancreatitis. Hypothesis. Emergency ERCP in gallstone pancreatitis (GSP) associated with jaundice or angiocholitis has a beneficial effect on pancreatic inflammation, eventually favoring laparoscopic cholecystectomy. We have compared 18 cases of emergency ERCP for GSP associated with jaundice or angiocholitis (Group 1) with 72 cases of GSP where ERCP was indicated on a selective basis (Group 2). Laparoscopic cholecystectomy was done after the acute bout of pancreatitis subsided. The minimally invasive approaches were completed in 89% cases in Group1 and in 90% cases in Group 2, respectively. Pancreatic morbidity was better in Group 1 (0% vs. 15.2%), but without statistical significance. General morbidity was significantly lower (0% vs. 23.6%) as well as the mean hospital stay (13 +/- 5.5 days vs. 17 +/- 10.4 days). Only the lower general morbidity and the shorter hospital stay assert the hypothesis that emergency ERCP in GSP associated with jaundice or angiocholitis has a beneficial effect on pancreatic inflammation, eventually favoring laparoscopic cholecystectomy. PMID- 15810705 TI - [Learning curve and his consequences in laparoscopic antireflux surgery]. AB - The laparoscopic fundoplication became the gold standard of the laparoscopic antireflux surgery (LARS). Our aim is to indicate the evolution of the learning curve as well as its consequences on the patient's outcome. We studied the gastro esophageal reflux (GER) cases treated laparoscopically in Coltea University Hospital throughout 6 years. We gathered a group of 40 patients with an average age of 54, 57 years and a sex ratio F:M = 1.67. The patients had either a simple GER disease, small and medium hiatal hernias (21 cases) or giant hiatal hernias (GHH--19 cases). Two equal groups resulted: group 1 consisted of the first 20 patients operated from 1999 to 2002, group 2 consisted of the rest of the patients. Operating time, hospital time, complication rate and postoperatory endoscopy were compared. The average of the operating time was calculated. For giant hiatal hernias, a separate average was also taken into account. The total operating time for GER, small and medium hiatal hernias was 115 min in group 1 and 80 min in group 2 meanwhile for GHH it was 143 min vs. 130 min. The average operatory time was 129 min vs. 105 min. All these differences were statistically significant but there were no differences concerning complication rate and post operatory endoscopy. Although the learning of the laparoscopic fundoplication requires practice, the learning curve does not have influence on the patients' outcome. PMID- 15810706 TI - [Laparoscopic cholecystectomy]. PMID- 15810704 TI - [Laparoscopic radical cystectomy with enteric urinary diversion. Alternative to open cystectomy?]. AB - Radical cystectomy is the standard therapy for invasive bladder cancer, with best oncological results compared to any other therapeutic alternative. Even if laparoscopic radical cystectomy (LRC) is a well established surgical procedure, performing the urinary diversion completely intracorporeal, is still a challenge due to technical difficulties and associated complications. The aim of our study is to present the first series of LRC with ureterosigmoidostomy. Since May 2004 were performed 7 LRC (5 males and 2 females) (Gr. A). All cases were T2N0 clinical stage. These patients were compared with a retrospective group of 50 patients who underwent open procedure (Gr. B) in term of: operative time, blood loss, analgesic requirements, and hospital stay. LRC has a longer operative time but with statistically significant lower blood loss, less analgesia, and shorter hospital stay. The resection margins of the surgical specimens were tumor free at pathologic examination. Long-term follow-up is pending. On short-term, the results of laparoscopic radical cystectomy are encouraging. When significant experience in laparoscopic surgery is lacking, surgeons should exercise caution with completely intracorporeal urinary diversion. PMID- 15810707 TI - [Spontaneous esophageal rupture in three patients]. AB - Boerhaave's syndrome represents the most severe perforation of the gastrointestinal tract. The typical clinical presentation is by a sudden onset of pain after a vomiting effort on the background of an alimentary and alcoholic abuse. It usually associates the cervicomediastinal emphysema, altogether forming the Mackler triad. The atypical presentation and the rarity of this entity usually lead to the delay in diagnosis in 50% of the cases. The diagnosis requires native and hydrosoluble contrast radiological examination which may reveal, altogether with the esomedistinal or esopleural fistula, pneumomediastinum, cervical subcutaneous emphysema, pleural effusion or hydropneumothorax. The early diagnosis and surgical treatment in the first 24 hours after the perforation offer the best chances for survival. We present and analyze the cases of 3 patients with spontaneous esophageal rupture with their different evolution and the diagnostic and treatment steps in each case. PMID- 15810708 TI - [Choledochal cyst as a complex malformation of the intra and extrahepatic bile ducts]. AB - The authors present the case of a 64 years woman with a choledochal cyst along with a complex malformation of both intra and extrahepatic bile ducts. The patient was admitted with a diagnosis of acute and underestimated cholecystitis, which was in fact a real acute severe suppurated angiocolitis located at the cystic cavity level along with severe hepato-renal failure. As long as the choledochal cyst evolves as a stand-alone entity, it can frequently be associated with other malformations of the bile ducts--such as choledochal duct stenosis or abnormal connection of the common hepatic duct with pancreatic ducts. The main dilatation was engulfing the whole common bile duct with a fusiform aspect, and the cranial end is opening into a long hepatic duct from which were emerging several biliary segments branches, for both hepatic lobes. This aspect of a multistage convergence with four branches which is opening into a long hepatic bile duct and then in the choledochal cyst, represents an unusual malformation which does not respect the rules described by former published authors. PMID- 15810709 TI - [Stromal tumor of Meckel's diverticulum--a rare source of gastrointestinal bleeding and a real diagnostic problem]. AB - Although Meckel's diverticulum is the most common congenital abnormality of the gastrointestinal tract, complications in adults are rare, especially in the elderly. Lower gastrointestinal bleeding as a result of Meckel's diverticulum with ectopic gastric mucosa is unusual among the elderly. The case we report involved a 69 year old man with gastrointestinal hemorrhage found to be due a stromal tumor developed on Meckel's diverticulum. We considered our case interesting because of its rarity and of preoperative diagnosis difficulty. PMID- 15810710 TI - A rare indication for liver resection. AB - We present the rare case of a young female with a right upper abdomen tumoral mass and suffering abdominal discomfort. A combination of ultrasonography, computed tomography, magnetic resonance imaging and laparotomy was utilized to conclude a diagnosis of Riedel's lobe. Laparotomy and a resection of Riedel's lobe were selected as the correct therapeutic solutions. PMID- 15810711 TI - [Reconstruction of cervical esophagus by using laparoscopically assisted jejunal loop harvesting and microsurgical transfer]. AB - For segmental cervical esophageal reconstruction, the free transfer of jejunal flap tends to become a standard procedure. Graft harvesting by laparotomy presumes increased morbidity at donor area level. There was described by now in the literature laparoscopic techniques for harvesting of jejunal graft, which have reduced the complications following laparotomy. We wish to present here an experimental model of laparoscopically assisted harvesting of jejunal flap used for reconstruction of cervical esophagus by free transfer. The harvesting of jejunal segment was achieved in 12 dogs; 4 cases subsequently underwent microsurgical free transfer. There are presented original contributions of authors. The harvesting average time was 115 minutes; the meantime graft ischaemia was 48 minutes in 4 cases with microsurgical transfer. Postoperative survival was 100%. Graft viability was achieved in 75% at 12 hours and 50% at 24 hours. No major operative accidents were recorded. We present a standardized experimental model, which could be an usefull guide in human therapeutics. PMID- 15810712 TI - [Complementary examinations]. PMID- 15810713 TI - [S.R.C activity--Bucharest chapter]. PMID- 15810714 TI - [Urinary incontinence: current status]. PMID- 15810715 TI - Pelvic floor disorders in women: an overview. AB - Urinary incontinence affects 10% to 58% of community-dwelling women and up to 50% of nursing home residents. Prevalence of incontinence appears gradually to increase during young adult life, has a broad peak around middle age, and then a steady increase in the elderly. It has been shown to affect a person's social, clinical, and psychological well-being. The majority of women with incontinence do not seek medical help. The estimated annual direct cost of urinary incontinence alone in women in the United States (in U.S. dollars) is 12.4 billion. The proportion of the United States population over age 75 was 22% in the year 1999 and is expected to grow substantially during the coming decades. Pelvic floor disorders such as urinary incontinence, fecal incontinence, and pelvic organ prolapse affect older women disproportionately. Because of this relationship, it is estimated that the growth in demand for services to care for female pelvic floor disorders will increase at twice the rate of growth of the population over the next 30 years. Approximately 10% of women have surgery for pelvic organ prolapse or urinary incontinence in their lifetime and nearly 30% of these operations are for recurrent disease. This fact suggests that failure rates after surgery for pelvic floor disorders are high, in spite of the introduction of many new surgical diagnostic techniques, tools and operations. Numerous techniques have been developed to evaluate the type and extent of urinary and fecal incontinence and prolapse. A number of treatment options exist, ranging from behavioral to medical and surgical approaches. This document will discuss some of the general issues related to pelvic floor disorders and suggest areas for future research. PMID- 15810716 TI - [Urinary incontinence and other pelvic floor damages: ethilogy and prevention strategies]. AB - Urinary incontinence, as well as additional pelvic floor damage, such as third and fourth degree muscular lacerations, as well as fecal incontinence, genital prolapse or dyspareunia, result from obstetric trauma, and are generally linked to the first delivery. The purpose of this study is to analyze, from a physiotherapeutic point of view, and therefore from the perspective of muscular physiology and biomechanics, why this damage occurs, while studying the birth process and the way it is currently performed in most hospitals in our country. Analysis of the birth process and, in short, of the different types of positions used for the first and second stage of labor, as well as of the care provided for women in the puerperium, leads us to propose a global prevention strategy to be carried out in three stages: --Ante-natal prevention: specific preparation of the pelvic floor and abdominal musculature during pregnancy, using massage techniques and manual stretching of the perineum. In addition, the pregnant woman learns these positions and methods of pushing, which makes the first and second stage of labour easier. An osteopathic treatment of the pelvis joints is performed in order to facilitate their mobility or to liberate blockades, if they exist. - Prevention during labour: During this stage, physiology is respected and manual, position-based and breathing techniques are implemented in order to enhance the protection of the baby and of the pelvic floor. --Postpartum prevention: The action is focused on the pelvic floor, through diaphragmatic and abdominal exercises or postures and, if necessary, osteopathic treatment in the early puerperium, in order to facilitate the correct involution of all soft tissues and the pelvic joints involved in labor. Early specific physiotherapeutic treatment will be proposed for women with functional pathology six weeks after delivery. PMID- 15810717 TI - [Female incontinence: urodynamic evaluation]. AB - OBJECTIVE: To evaluate the functional studies in women with stress urinary incontinence. METHODS: Emphasis is placed on comprehensive understanding of stress urinary incontinence in women as well as Urodynamic techniques and findings which apply to women with this condition. RESULTS: Although prospective randomized controlled trials to evaluate the clinical efficacy, cost effectiveness and effect on quality of life of a pre-operative Urodynamic assessment are necessary, to date only Urodynamic Investigation (Non-invasive uroflowmetry, filling cystometrogram, valsalva leak point pressure, pressure-flow studies and urethral profile) provide enough information for treatment decisions and prognosis in cases of female urinary incontinence. PMID- 15810719 TI - [Surgery for urogenital prolapse]. AB - Urogenital prolapse can have a significant impact on quality of life. As the population continues to age, the prevalence of urogenital prolapse is increasing, and the lifetime risk of requiring surgery for urogenital prolapse or incontinence is now approximately 11%. The majority of women presenting with symptomatic prolapse suffer from multiple defects of pelvic support and require comprehensive repair to relieve symptoms. An understanding of normal pelvic support structures provides the basis for the anatomic approach to repair. Many appropriate options exist for surgical correction of urogenital prolapse. Procedures to reestablish apical support include culdoplasty techniques, uterosacral ligament suspension, sacrospinous suspension and colpopexy. Repair of the anterior compartment can be achieved with colporrhaphy and paravaginal repair. Posterior compartment defects are repaired with colporrhaphy, site specific rectovaginal repair and perineorrhaphy. Most often, surgical correction of urogenital prolapse can be performed vaginally, which avoids the risks associated with laparotomy. Laparoscopic approaches for apical support and paravaginal repair may reduce the risks associated with laparotomy, but long-term follow-up data are not yet available with these techniques. The use of graft reinforcement for anterior and posterior repairs may offer improved success rates, particularly in patients with recurrent prolapse. However, further outcome studies are needed and the risks associated with the use of mesh must be considered. PMID- 15810718 TI - [Urinary incontinence: anticholinergic treatment]. AB - The bladder and urethra are a functional unit controlled and coordinated by the central and peripheral nervous systems. Voluntary or involuntary contraction of the Detrusor muscle depends mainly on muscarinic receptor stimulation by Acetylcholine, which is released from parasympathetic terminals. This receptors are the antimuscarinic (anticholinergic) drug target. Anticholinergic drugs are the first line of treatment for overactive bladder. They have been widely investigated and have shown superior efficacy to placebo in numerous studies. Their effects on bladder function result in an increase of the capacity at which involuntary detrusor contractions are produced, a decrease in its amplitude and an increase in bladder capacity. The purpose of this Literature Review is to provide an up-to-date overview of the current situation of this form of treatment (Oxybutinin, Long Acting Tolterodine, Trospium Chloride) and the main developments in this area. Some of these drugs are already commercially available in Spain (Long Acting Tolterodine), some may appear soon (Extended Release Oxybutinin, Transdermal Oxybutinin), and others are available for specific cases (Intravesical Oxybutinin). Other drugs, which are theoretically more uroselective (Darifenacin or Solifenacin), are not yet freely available on the open market, while yet others are already in use in other countries (Propioverine). Finally I would like to mention the increasing use of Botulinum Toxin detrusor injection for bladder hyperactivity that fails to respond to conservative treatments. PMID- 15810720 TI - [Urinary incontinence surgery: TVT procedure]. AB - The T.V.T. procedure has become a technique for Urinary incontinence and its different presentations. Many trials have reported the efficacy of this technique for different situations, as well as its results and complications. We review the literature and analyze the results and complications, comparing this technique with other such procedures. PMID- 15810721 TI - [Female urinary stress incontinence: transobsturator technique]. AB - The wide use of retropubic TVT has been associated with various complications. To avoid these, alternative procedures have been developed and continence rates obtained with these new routes have been quite similar to those after classic TVT. In the transobsturator technique (TOT) described by Delorme and colleagues in 2001, the tape is inserted through the obturator foramens from outside to inside and is positioned without tension under the urethra. Another surgical technique allows the passage of a tape from inside to outside. The aim of this paper is to describe a new, simple surgical technique for the treatment of female urinary stress incontinence and to evaluate its feasibility. PMID- 15810722 TI - [The triple perineal operation with prosthesis. First 100 cases results]. AB - The authors describe the technique of Triple Operation for Prolapses with Prostheses for the surgical cure of genital prolapse using a recto-vaginal interposition prosthesis for cystocele, rectocele and suspension of the vaginal dome by a posterior retro- and trans-levator muscle strip. The preliminary results over a continuous series of 100 patients operated on between June 2001 and June 2003 showed perioperative complications (7 bladder injuries) and postoperative complications (4 vaginal erosions related to the prosthesis including one that was secondarily infected, and a hematoma with secondary abscess of the pararectal fossa which required the ablation of the material). As far as anatomy was concerned, the results were excellent for the correction of hysterocele, apical prolapse and rectocele. On the functional level, dyspareunia was noted. Six of the ten preoperative mechanical dyschesias were considerably improved. PMID- 15810724 TI - [Life years, memory, and hope]. PMID- 15810723 TI - [Neurostimulation and neuromodulation in urinary incontinence]. AB - Electric stimulation in Urology has undergone considerable development since the 1970s. The implantation methods for urinary system stimulation are used for bladder emptying in neurogenic bladders (Neurostimulation) or the modulation of the electrical transmission of the voiding cycle caused by chronic detrusor sphincter dysfunction (Neuromodulation). Both techniques have in common the direct stimulation of the sacral roots, responsible for the urinary reflex arc. The first of them works via external stimulation over an implanted antenna, and the second via permanent stimulation through an implanted pacemaker. We review in this article the indications, technique and results of both therapies, specially focused on the Spanish experience. PMID- 15810725 TI - [Journal of Applied Biochemistry and Microbiology is 40 years old]. PMID- 15810726 TI - [Chitosan-based polyelectrolyte complexes: a review]. AB - The review focuses on the formation of polyelectrolyte chitosan complexes with biologically active compounds and prospects of use thereof. The possibility of obtaining low-molecular-weight, water-soluble batches of chitosan, which differ in their degree of acetylation, is discussed, with emphasis on their use for binding nucleic acids into complexes. PMID- 15810727 TI - [Inhibition of soybean urease by polycarbonyl compounds]. AB - Competitive inhibition of soybean urease was studied at 36 degrees C in aqueous solution (pH 4.95) in the presence of polycarbonyl compounds (PCCs): oxalyldihydrazide (ODH), its polydisulfide (poly(DSODH)), three cyclic beta triketones (CTKs), and seven cyclic PCC species of differing structure. The inhibition constants of ureolysis (Ki) varied in the range 8.5-3800 microM depending on the structure of organic chelators for the nickel atom in urease. It was shown that pH variation within the range from 3.85 to 7.40 exerted a strong effect on the values of Ki] of three CTKs and hydroxyurea, which was used as a reference: pH dependences of lgK(i) were linear in all cases and displayed a break at pH 6.0-6.5. The most effective inhibitor of ureolysis was poly(DSODH), which contained approximately 28 carbonyl groups in the polymer molecule. The role of such factors as the number of carbonyl groups per PCC molecule, mutual arrangement, and reaction medium pH in the efficiency of the process of urease inhibition is discussed. PMID- 15810728 TI - [Effect of flavonoids of different structure on peroxidation of neutral lipids of animal origin]. AB - The effect of flavonoids of different structure (baikalein, baikalin, quercetin, dihydroquercetin, genistein, and daidzein) on the process of formation of lipid hydroperoxides during thermoinduced autooxidation of neutral lipids of animal origin. The minimum inhibitory concentration of isoflavones was found to be equal to 10(-3) M. Effective inhibitory concentration of other flavonoids (except baikalin) was equal to 10(-4) M. Baikalin was an effective promoter of lipid peroxidation. The antioxidant activity of the flavonoids tested was calculated taking ionol as a reference. PMID- 15810729 TI - [Effect of carbohydrates and polyols on amide content and protein fragmentation in a lactoglobulin preparation]. AB - Certain carbohydrates and polyols are used at various stages of the production of immunobiological preparations as stabilizers of biological activity, particularly in the production of lactoglobulin (against opportunistic pathogens) using membrane ultrafiltration. This study concerns the effect of these substances on changes in the amide content in proteins of this lactoglobulin. Lactoglobulin was incubated in near-physiological (0.9% NaCl, pH 5.5) 10% solutions of glucose, fructose, and sorbitol at 4 and 35 degrees C for 7, 14, and 28 days. A lactoglobulin solution in 0.9% NaCl, pH 5.5, was used as the control. All substances studied suppressed the reduction of the amide group content of asparagine and, in contrast, increased the rate of amide group removal from glutamine residues in proteins of lactoglobulin preparations. PMID- 15810730 TI - Hymeniacidon perleve associated bioactive bacterium pseudomonas sp. NJ6-3-1. AB - Among the marine bacteria isolated from cytotoxic sponge Hymeniacidon perleve, one strain NJ6-3-1 classified as Pseudomonas sp. showed both cytotoxic and antimicrobial activities. Fatty acid analysis indicated that the bacterial strain consists mainly of C16:1, C16:0, C18:1, C18:0, C15:0, C14:0. One unusual 9,10 cyclopropane-C17:0 fatty acid, and C26:0 also constitute as major component as well as the existence of squalene, the precursor of triterpenoids. The major metabolites in the culture broth were identified as alkaloids, including diketopiperazines and indole compounds, namely 3,6-diisopropylpiperazin-2,5 dione, 3-benzyl-3-isopropylpiperazin-2,5-dione, 3,6-bis-(2-methylpropyl) piperazin-2,5-dione, indole-3-carboxaldehyde, indole-3-caroxylic acid methyl ester, indole-3-ethanol, and quinazoline-2,4-dione. PMID- 15810731 TI - [Isolation and characterization of the thermostable DNA polymerase of the hyperthermophilic archaeum Thermococcus litoralis Sh1AM]. AB - Overall, 30 strains of hyperthermophilic archaea, representing seven species of the genera Thermococcus, Desulfurococcus, Thermoproteus, and Acidilobus, were tested for the presence of thermostable DNA polymerases. Thermostabilities of the polymerases varied distinctly among the strains within one species. Polymerases of five strains retained 60-100% activity upon incubation of the preparations at 95 degrees C for 120 min. A new DNA polymerase was isolated from the strain Thermococcus litoralis Sh1AM, possessing the enzyme with the most promising properties, and characterized. Molecular weight of the enzyme is 90-100 kDa. The purified DNA polymerase preserved 50% of the initial activity upon incubation at 95 degrees C for 120 min. The polymerase isolated displayed an associated 3'-5' exonuclease activity. The error rate when extending DNA strand was at least twofold lower compared with Taq polymerase. The main physicochemical and enzymatic properties of the new polymerase are similar to the known DNA polymerases of family B. PMID- 15810732 TI - [Bioconversion of phenyllactate to phenylalanine by bacteria of the genus Pseudomonas]. AB - The nature of phenylalanine auxotrophy in Pseudomonas sp. strain M9 was studied. A metabolic defect was found after the aromatic amino acid pathway branches from chorismate, so that the latter could not be converted to phenylpiruvate. It was shown that enzymatic activity of the strain could convert phenyllactate to phenylalanine. The reaction mechanisms were studied, and approaches to increasing the rate of the process and phenylalanine yield were proposed. PMID- 15810733 TI - [Sensitivity of various escherichia coli strains to 2,4,6-trinitrotoluene]. AB - The sensitivity of Escherichia coli strains K-12 and 055 to 2,4,6-trinitrotoluene (TNT) was found to correlate with the structural and functional properties of the outer lipoprotein membrane. The protective ability of the membrane of strain 055 is much lower than that of K-12. This is the cause of the greater sensitivity of 055 to the toxic action of TNT. High TNT concentrations (100-200 mg/l) suppressed the growth of 055, whereas K-12 grew at all TNT concentrations studied. Both strains adapted to high TNT concentrations by converting it by either nitroreduction or denitritation depending on concentration. The denitritation system of strain 055 started TNT degradation earlier than that of K-12. PMID- 15810734 TI - [Use of claydite-immobilized oil-oxidizing microbial cells for purification of water from oil]. AB - Oil-oxidizing bacteria were isolated from oil-polluted soil and water samples and identified as Acinetobacter calcoaceticus K-4, Nocardia vaceinii K-8, Rhodococcus erythropolis EK-1, and Mycobacterium sp. K-2. It was found that immobilization of the bacteria on an expanded clay aggregate accelerated their growth and consumption of hydrocarbon substrates. It was also found that water polluted with 100 mg/l oil could be purified with Rhodococcus erythropolis EK-1 and Nocardia vaceinii K-8 cells immobilized in this way. The dependence of the degree of water purification on its flow rate, aeration, and availability of nitrogen and phosphorus sources was determined. The efficiency of water purification from oil by immobilized Rhodococcus erythropolis EK-1 cells at high flow rates (of up to 0.68 l/h), low aeration (of 0.1 l/l per min) and an intermittent supply of 0.01% diammonium phosphate reached 99.5-99.8%. PMID- 15810735 TI - [Evaluation of substrate specificity of biosensor models based on strains degrading polycyclic aromatic compounds]. AB - Models of microbial biosensors based on 11 strains of degrading surface-active substances (SASs) and polycyclic aromatic hydrocarbons (PAHs) were studied. Substrate specificity, sensitivity, and stability of biosensor models were comparatively evaluated. PMID- 15810736 TI - [Formation of structured communities by natural and transgenic naphthalene consuming bacteria]. AB - This study concerns the formation of structured communities by pure cultures and binary associations of Pseudomonasfluorescens transgenic strains and natural heterotrophic bacterial species in naphthalene-containing media with various osmotic pressures. It was shown that cells of P. fluorescens strain 5RL, harboring a recombinant construct in the chromosome, were more resistant to the combined action of the stress factors under study than P. fluorescens 82/pUTK21, harboring a recombinant construct within a plasmid. Natural P. fluorescens 1 strains, particularly Vibrio sp. 14, were more viable at high osmotic pressures and naphthalene concentrations. Experiments with the combined introduction of transgenic and natural bacterial strains at high osmotic pressures demonstrated the stable coexistence of bacterial associations in biofilms, independent of naphthalene concentration. Strains considered for introduction into the environment for bioremediation should be assessed with regard to their susceptibility to the combined effect of anthropogenic and natural stress factors. The design of bacterial associations for the same purpose should take into account the effect of factors important for their survival in polluted areas. PMID- 15810737 TI - [Fermentation of a high-protein plant biomass by introduction of lactic acid bacteria]. AB - Lactic acid bacteria displaying increased ability to produce lactic acid, medium proteolytic activity, and tolerance to osmotic stress were isolated under selective conditions from phyllosphere and rhizosphere of registered and raised cultivars of legumes. Lactic fermentation of poorly ensilable leguminous plants (red clover and Caucasian goat's rue) was performed by introduction of rifampin resistant homofermenting representatives of the genus Lactobacillus (selected according to a set of technologically important characteristics). The results demonstrate that introduction of active local strains of lactobacteria, as well as the collection strain Lactobacillus plantarum BS 933, enhances activation of ensiling and increases the quality of fodder, as assessed according to the standard criteria (a decrease in pH of the medium, the ratio of lactic acid to fatty acid homologues, and the composition of silage microflora). PMID- 15810738 TI - [Trigliceropeptides from pseudomonades: new agents for biological control of phytopathogenic fungi]. AB - Substances of peptide nature have been isolated from low-molecular-weight fraction of extracellular metabolites of Pseudomonas chlororaphis (strains IB 51 and IB 6) and Pseudomonas putida (strain IB 17). These substances possess antifungal activity against a broad range of phytopathogens but lack the properties of siderophores. It has been shown by HPLC, IR, and 13C NMR spectroscopy that isolated metabolites are glycerol tripeptides with molecular weights in the range 2.8-3.0 kDa. PMID- 15810739 TI - [Production of mycophenolic acid by fungi of the genus Penicillium link]. AB - Out of 36 strains of fungi of the genus Penicillium, some of which were isolated from ancient permafrost soils, 14 strains synthesized mycophenolic acid (MPA). Maximal (over 500 mg/l) accumulation of MPA in culture liquid was observed in P. brevicompactum strains (VKM F-457, VKM F-477, and VKM F-1150). This was the first study to detect MPA in representatives of the species P. rugulosum; in three strains of this species (VKM FW-665, VKM FW-717, and VKM FW-733), the level of MPA accumulation exceeded 300 mg/l. The time course of the synthesis of MPA by the P. rugulosum strain VKM FW-733 was studied. It was shown that the synthesis of this metabolite was dramatically intensified at the stationary growth phase (ten days). PMID- 15810740 TI - [Proteolytic enzymes as markers of productivity and heterosis of silkworm]. AB - A positive correlation between the activity level of cysteine proteinases in developing eggs of common silkworm moth (Bombyx mori L.), on the one hand, and a set of commercial characteristics, on the other, was found. This allows the determination of cysteine proteinase activities (pH optima of 3.0, 3.6, and 8.6) to be recommended as a biochemical test for an early prediction of potential productivity of silkworm breeds. A positive correlation between the activity level of acid cysteine proteinases in eggs of parental breeds and a set of commercial characteristics of their hybrids was detected, indicating a principal possibility of predicting the degree of heterosis. PMID- 15810741 TI - [Effects of natural and artificial defoliations on the content and composition of extractive substances in birch leaves]. AB - Qualitative and quantitative compositions of extracts of birch (Betula pendula Roth.) leaves after natural and artificial defoliations were studied. Composition of the fraction of total lipids was determined. Overall, 11 fatty acids were identified. Differences between the fatty acid compositions of total lipids in the trees subjected to defoliation, consisting in the increase in quantities of short-chain saturated fatty acids and trienoic acids, were detected. Nine individual compounds--six flavones, two flavanones, and one flavanonol--were isolated from the flavonoid fraction by column chromatography. It was found that the total content of extractive substances in birch leaves as well as amounts of free sterols, triterpene compounds, and flavones decreased 1 year after an artificial defoliation and 1 month after depredation of 75% of birch stands by gypsy moth. On the contrary, the contents of flavanones and flavanonol increased. The assay method proposed may be used for studying the compositions of plant extracts. PMID- 15810742 TI - [Formation of phenol compounds in various cultivars of wheat (Triticum aestivum L.)]. AB - The formation of soluble phenol compounds, including flavonols, was studied in winter (Erythrospermum, Lutescens 230, and R 47-28) and spring cultivars (Lada) of wheat (Triticum aestivum L.). The contents of soluble phenol compounds and flavonols were 1.8-2.6 and 0.5-1.3 mg/kg fresh weight, respectively. These results illustrate the similarity of phenol metabolism in leaves of winter and spring wheat cultivars. The exception was the cultivar R 47-28 that accumulated the maximum amount of phenol compounds (e.g., flavonols). In this cultivar the ratio of flavonols reached 50% of total soluble phenol content. In other cultivars, this parameter did not exceed 25-35%. The data indicate that the cultivar R 47-28 differs from other wheat cultivars in the metabolism of phenol compounds. The observed differences are probably related to genetic modifications of the cultivar R 47-28 during selection. PMID- 15810743 TI - [Role of L-phenylalanine ammonia lyase in the induced resistance and susceptibility of potato plants]. AB - Biogenic elicitors (chitosan and its complex with salicylic acid) and an immunosuppressor (laminarin) were shown to increase the activity of L phenylalanine ammonia lyase (EC 4.3.1.5) and protein synthesis in potato tubers. Laminarin did not decrease L-phenylalanine ammonia lyase activity. It is unlikely that the activity of this enzyme may serve as a criterion for the induced resistance. PMID- 15810744 TI - [Protein markers of traits of plants in breeding spring wheat for grain productivity and quality]. AB - A comparative study of the composition of gliadin proteins contained in grains of different-quality wheat cultivars Rollo and Drott and four forms of hybrids F9 and F10, obtained by crossing these cultivars, has been performed. The traits of grain productivity of ears, quality of flour, dough, and gluten were analyzed. Groups of genetically linked gliadin components, controlled by chromosomes 1 and 6 of homoeologous groups, have been identified as protein markers of selectively valuable traits of plants. PMID- 15810745 TI - User and carer participation in research in palliative care. PMID- 15810746 TI - Breaking bad news: experiences, views and difficulties of pre-registration house officers. AB - OBJECTIVES: To obtain information regarding the involvement of pre-registration house officers (PRHOs) in the discussions on bad news, and the competency and difficulties they perceive in clinical practice. DESIGN: Structured telephone interviews. PARTICIPANTS: 104 PRHOs. MAIN OUTCOME MEASURES: Information about frequency and quality of involvement of PRHOs in discussions on bad news with patients and relatives, perceived competency and difficulties related to this task as well as ethical views concerning the disclosure of bad news. RESULTS: 82 PRHOs (78.9%) had initiated the breaking of bad news to a patient at least once, whilst patients themselves had initiated discussions of bad news by asking the doctors questions (92.3%). Almost all (96.2%), indicated that they had broken bad news to relatives of a patient. The majority of the junior doctors participating in our study felt fairly or very confident (90.4%) to break bad news. 'Often' quoted difficulties for over a fifth of the sample included 'Thinking I was not the appropriate person to discuss the bad news', 'Having all the relevant information available', 'Dealing with emotions of patient/relative', 'Lack of privacy' and 'Patients/relatives do not speak English'. Although 99 PRHOs (95.2%) believed that patients should be informed about a serious life threatening illness, 30.8% of the participants stated that doctors need to judge whether or not to tell a patient bad news. Factors most frequently selected by the PRHOs from a given list of possible factors contributing to a gap between theory and practice included problems with the organization of clinics (73.1%), insufficient postgraduate training (63.5%) and lack of staff (54.8%). CONCLUSIONS: The results indicate that PRHOs are frequently involved in the breaking of bad news. Whilst no claims can be made for their,actual performance in practice, their perceptions of competency would indicate that the extensive and compulsory undergraduate teaching they had received on this subject has served to prepare them for this difficult task. Organizational and structural aspects need to be taken into account as factors assisting or undermining doctors in their efforts to put into practice ethically sound and skilled communication when disclosing bad news. PMID- 15810747 TI - Effects of spiritual care training for palliative care professionals. AB - Little is known about the effects of spiritual care training for professionals in palliative medicine. We therefore investigated prospectively the effects of such training over a six-month period. All 63 participants of the three and a half-day training were asked to fill out three questionnaires: before and after the training, as well as six months later. The questionnaires included demographic data, numeric rating scales about general attitudes towards the work in palliative care, the Self-Transcendence Scale (STS), the spiritual subscale of the Functional Assessment of Chronic Illness Therapy (FACIT-Sp) and the Idler Index of Religiosity (IIR). Forty-eight participants (76%) completed all three questionnaires (91% women, median age 49 years; 51% nurses, 16% hospice volunteers, 14% physicians). Significant and sustained improvements were found in self-perceived compassion for the dying (after the training: P=0.002; 6 months later: P=0.025), compassion for oneself (P <0.001; P =0.013), attitude towards one's family (P =0.001; P =0.031), satisfaction with work (P < 0.001; P = 0.039), reduction in work-related stress (P < 0.001; P = 0.033), and attitude towards colleagues (P = 0.039; P = 0.040), as well as in the FACIT-Sp (P < 0.001; P = 0.040). Our results suggest that the spiritual care training had a positive influence on the spiritual well-being and the attitudes of the participating palliative care professionals which was preserved over a six-month period. PMID- 15810748 TI - Social class and access to specialist palliative care services. AB - OBJECTIVE: To determine any social class differences in place of death of cancer patients in South Bristol; to explore the experience of carers; and to identify inequalities in access to palliative care. DESIGN: Two-part study: (1) A cross sectional survey of all 960 cancer deaths in South Bristol between September 1999 and December 2002. (2) A qualitative in-depth interview study of 18 carers of patients who died of cancer during the same period in South Bristol. Fourteen of those who died were from social class IIIM (manual), IV and V (i.e. lower social classes). MAIN OUTCOMES: Place of death of patients according to social class and geographical distance from the hospice. Carers' accounts of the way in which illness and death were conducted, and their response to the management of death and dying. RESULTS: The cross sectional survey showed that patients from social class V were less likely to die in the hospice. This finding was independent of geographical proximity. In the qualitative study, no class specific beliefs about death and dying were identified. Attitudes to the way dying should be conducted were common across the classes. Families expected to be present and centre stage at the time of death and for it to be conducted in a dignified and personal manner. Health care staff in all settings supported them in this aim. No one in this study died without a family member present. Some elderly carers were less open than younger carers in the way they talked about death and did not wish to be present at the death. Some carers from social class IV and V were less active in seeking information or asking for hospice admission than carers from other classes. Unrealistic expectations about the availability of hospice beds were common to all carers. Anxiety was common among carers. It was reduced by the provision of reliable and consistent healthcare support, by information provided in a timely and sensitive way, and by open and shared decision making between carer and patient. Most important in reducing anxiety was the support of a second carer who lived locally and was reliable. Carers from social classes IIIM (manual), IV and V were more likely to have this kind of support than carers from social classes I, II and IIIN (non-manual). CONCLUSION: Although cancer patients from social class V were less likely than others to die in the hospice, social inequality in access to or utilisation of healthcare in terminal illness was not prominent in carers' accounts. When it did arise, it was associated with passivity in seeking information and support on the part of some carers from social classes IV and V. Carers from social classes IIIM-V received more regular and reliable support from their families than those from social classes I-IIIN. PMID- 15810749 TI - General practitioners (GPs) and palliative care: perceived tasks and barriers in daily practice. AB - BACKGROUND: General practitioners (GPs) play a crucial part in palliative care. The quality of care can be improved by investigating and addressing barriers perceived by GPs in daily practice. The aim of this study was to investigate GPs' task perception and barriers involved in palliative care. METHODS: Qualitative focus group study. We gathered together a group of GPs representing a broad range of experience in palliative care. Content analysis was performed to derive a comprehensive view of tasks and barriers in daily palliative care. RESULTS: GPs described their palliative care tasks as satisfactory and varied, but burdensome. Palliative care tasks included somatic and psychosocial care. Opinions differed with respect to whether the coordination of care belonged to the primary GP tasks. Barriers were classified according to three levels: (1) personal: barriers related to knowledge, skills, emotions; (2) relational: barriers concerning communication and collaboration; (3) organizational: barriers related to the organization of care and compartmentalization in healthcare. CONCLUSIONS: This study revealed a complex web of tasks and barriers. It may be possible to trace back a problem (lack of knowledge, for example) on the personal level to an isolated knowledge gap, but the problem may well have originated from communication or compartmentalization problems. To maintain GPs' feeling of being at ease with palliative care requires helping them acquire the appropriate balance between technical and organizational interventions and a compassionate orientation to their terminally ill patients. PMID- 15810750 TI - Attitudes of Danish doctors and nurses to palliative and terminal care. AB - BACKGROUND: The WHO definitions of palliative care have been adopted in Denmark and implemented in The National Guidelines from 1999, but service developments have been very slow and not according to the recommendations. Attitudes to palliative care of Danish doctors and nurses may in part account for this. OBJECTIVE: To assess the attitudes to issues related to palliative care of doctors and nurses in a Danish county hospital and the related primary care services. DESIGN: Cross-sectional survey using a mailed, self-administered questionnaire answered anonymously. PARTICIPANTS: Nurses and doctors employed in a county hospital in Denmark, homecare nurses and general practitioners from the related primary care services. OUTCOME MEASURES: The responses from the groups were compared by chi2 statistics (where ordinal variables with chi2 for trend). Data were analysed using SPSS 10.0. RESULTS: 347 responded, response rate 76%. Eighty-one per cent of all respondents were currently caring for terminally ill patient(s), 94% had done so within the last six months. Hospital doctors see more terminally ill patients than GPs (P = 0.002). Comparison of doctors (both hospital and GPs) with nurses showed that nurses were more likely to definitely agree that palliative/terminal care was a rewarding part of their work (61% 'definitely agree' versus 30%), and they were less likely to prefer to leave care of these patients to others (4% 'definitely/probably agree' versus 9%). Nurses reflected more on existential matters (80% 'definitely/probably agree' versus 63%) and were more likely to agree that dealing with a dying patient made them aware of their own feelings regarding death (97% 'definitely/probably agree' versus 80%). Only 7% of all respondents reported 'being an active member of a religious community'. Ninety-two per cent of all respondents agreed that doctors play a key role in reducing the suffering of dying patients, but 59% of nurses versus 9% of doctors 'definitely/probably agree' that 'it is primarily the task of nurses to deal with patients reactions to death'. There were significant differences between hospital doctors and GPs, with the former less likely to agree that palliative and terminal illness is rewarding, more likely to leave care of dying patients to others, and more likely to 'probably' or 'definitely agree' that it is more satisfying to work with patients who will improve. Home care nurses reflected more on existential matters than their hospital colleagues, and were more likely to 'definitely agree' that palliative/terminal care is rewarding. Differences between groups seemed to be due to profession (doctor versus nurse) and setting (hospital versus community) rather than age or gender. CONCLUSION: These findings suggest that in Denmark nurses demonstrate more positive attitudes to the care of palliative/terminally ill patients than doctors, and that attitudes amongst doctors and nurses working in the community are more positive than those of the colleagues in hospitals. There is currently little education in the principles and practice of palliative care in Denmark. These findings will inform the development of appropriate palliative care education for doctors and nurses working both in the hospital and in the community in Denmark. They also raise the possibility that part of the inertia in the development of palliative care in Denmark is related to the lack of education and, in particular, to the need of support for doctors and nurses providing terminal care so they are enabled to be more reflective on the care they currently provide. There is evidence that education in palliative care can change health professionals' attitudes to palliative and terminal care, and this now needs to be investigated in Denmark. PMID- 15810751 TI - Low dose diamorphine reduces breathlessness without causing a fall in oxygen saturation in elderly patients with end-stage idiopathic pulmonary fibrosis. AB - There is very little evidence regarding the safety and efficacy of opioids for the control of dyspnoea in the terminal stages of idiopathic pulmonary fibrosis (IPF). We conducted an open case series study of 11 elderly opioid-naive patients referred for management of severe breathlessness before and after their first injection of 2.5 mg diamorphine subcutaneously. Subjective breathlessness, measured by a 100 mm visual analogue scale, fell by a mean of 47 mm in the first 15 min (P < 0.0001) and the mean heart rate fell by 12/min (P = 0.007). There were small non-significant falls in the mean respiratory rate (2/min), systolic blood pressure (6 mmHg) and oxygen saturation (1%). These changes were maintained at 30 min. Follow up treatment with oral morphine remained effective in reducing the symptom of breathlessness and no patient showed signs of respiratory depression. Low dose opioids are effective and safe in the palliative management of IPF in frail elderly patients. PMID- 15810752 TI - Stability of morphine sulphate and diamorphine hydrochloride in intrasite gel. AB - Several studies have reported that opioids applied topically to painful ulcers produce an analgesic effect. It is unknown whether these opioids (usually mixed with hydrogels) are stable and, if so, for how long. We investigated the stability of morphine sulphate and diamorphine hydrochloride, each mixed with intrasite gel at a concentration of 1.25 mg/mL. Samples were prepared in the laboratory and then stored in plastic containers in the dark, at room temperature, in conditions of normal day/night at room temperature, and at 4 degrees C. Aliquots were collected from each container over a 28-day period and analysed using HPLC. No known degradation products were measured in the morphine intrasite gel mixture stored for up to 28 days, irrespective of the temperature and whether or not samples were exposed to light, suggesting that morphine remains stable. Diamorphine, breaks down to morphine and no other degradation products are measurable. PMID- 15810753 TI - Palliative care in children in Wales: a study of provision and need. AB - AIM: To establish incidence and prevalence of children needing palliative care in Wales. PATIENTS AND METHODS: Children were identified in three ways: (1) from paediatricians using the Welsh Paediatric Surveillance Unit (WPSU); (2) referrals to the specialist palliative medicine service based in Cardiff; and (3) children under the care of the two principal children's hospices serving Wales. All children referred or reported between January 2001 and December 2002 were included. RESULTS: A total of 226 children were identified. Fifty (22%) were identified by paediatricians, 58 (26%) were referred to the specialist paediatric palliative medicine service, 158 (70%) had been under the care of a children's hospice, and 34 (15%) were identified by more than one source. This study identified approximately 3.75 per 10000 children. This is about half the prevalence figures quoted in the ACT/ RCPCH document in 1997. CONCLUSIONS: The study may underestimate prevalence. Children needing palliative medicine are still under-recognized in Wales. The overlap between children's hospice care and specialist paediatric palliative medicine is relatively small. PMID- 15810754 TI - Interpersonal processes in palliative care: an attachment perspective on the patient-clinician relationship. AB - The progression of advanced disease often brings enormous adaptational challenges related to a loss of the capacity to be self-sufficient and to a heightened need for support from others. The increasing need to rely on others at the end of life may reactivate relational problems and may trigger threatening feelings of vulnerability and dependency. Clinicians may find it difficult to identify and respond appropriately to the specific relational needs of individuals receiving end-of-life care. Attachment theory is a developmentally-based approach to understanding the formation and maintenance of relationships relevant to felt security. Although attachment theory has been widely applied to parent-child relationships and to relationships of individuals with chronic medical conditions to their medical caregivers, it has not previously been applied to the understanding of clinician-patient relationships in the palliative care setting. This paper provides an overview of this theory and demonstrates its application and relevance to palliative care. PMID- 15810755 TI - Physician-assisted death: attitudes and practices of community pharmacists in East Flanders, Belgium. AB - This study investigates attitudes and practices of community pharmacists with respect to physician-assisted death. Between 15 February and 15 April 2002, we sent anonymous mail questionnaires to 660 community pharmacists in the eastern province of Flanders, Belgium. The response rate was 54% (n = 359). Most of the pharmacists who responded felt that patients have the right to end their own life (73%), and that under certain conditions physicians may assist the patient in dying (euthanasia: 84%; physician-assisted suicide: 61%). Under the prevailing restrictive legislation, a quarter of the pharmacists were willing to dispense lethal drugs for euthanasia versus 86% if it were legalized, but only after being well informed by the physician. The respondents-favour guidelines for pharmacists drafted by their own professional organizations (95%), and enforced by legislation (90%) to ensure careful end-of-life practice. Over the last two years, 7.3% of the responding pharmacists have received a medical prescription for lethal drugs and 6.4% have actually dispensed them. So we can conclude that community pharmacists in East Flanders were not adverse to physician-assisted death, but their cooperation in dispensing lethal drugs was conditional on clinical information about the specific case and on protection by laws and professional guidelines. PMID- 15810756 TI - The Cochrane pain, palliative and supportive care group: current reviews and work in progress. PMID- 15810757 TI - The association of marital status and hospice use in the USA. AB - BACKGROUND: Married individuals are more likely to use hospice than unmarried individuals. We examine this association and how it is influenced by gender. METHODS: Medicare beneficiaries dying of cancer were studied. RESULTS: Currently married (OR 1.36 95% C.I. 1.28-1.45) or ever married (OR 1.23 95% C.I. 1.16-1.31) subjects were more likely to use hospice than never married subjects. A significant interaction between marital status and gender (P < .001) was observed. CONCLUSIONS: Subjects likely to enroll in hospice are subjects likely to have greater supportive relationships. PMID- 15810758 TI - Tamoxifen for retroperitoneal fibrosis. AB - Retroperitoneal fibrosis is characterised by diffuse inflammatory proliferation of fibroblasts and the deposition of collagen fibrils throughout the retroperitoneum, especially in perivascular sites. This results in blockage of the ureters and other tubular retroperitoneal structures such as the vasculature and lymphatic channels. Left untreated, it typically follows a progressive and fatal course. The condition responds favourably to tamoxifen and several case reports describe an improvement in renal function on this therapy. This case report describes a subjective and objective functional improvement of lymphoedema secondary to retroperitoneal fibrosis with the use of tamoxifen. PMID- 15810759 TI - NSAIDs: gastroprotection or selective COX-2 inhibitors? PMID- 15810760 TI - Autopsy in palliative medicine. PMID- 15810761 TI - Caution with nalbuphine in patients on long-term opioids. PMID- 15810763 TI - Psychogenic delirium? PMID- 15810762 TI - Prescribing palliative oxygen: a clinician survey of expected benefit and patterns of use. PMID- 15810764 TI - Patient choice regarding place of death. PMID- 15810765 TI - A prospective study on changes of cognitions, interpersonal influences, and physical activity in Taiwanese youth. AB - The decline of physical activity during adolescence is not well understood. This prospective study followed 969 Taiwanese students from eighth to ninth grade and examined their physical activity-related cognitions, interpersonal influences, and behavior of physical activity across the grade transition. Gender differences were found in the effects of the grade transition on physical activity and its related cognitions. The correlation coefficients showed moderate tracking for physical activity, cognitions, and interpersonal influences for both genders. The results also indicated that cognitions are more stable than behavior. The present study provides evidence of the decline in physical activity in Taiwanese youth across the grade transition as well as gender differences in the variables cognitions and interpersonal influences. PMID- 15810766 TI - Evaluating the sustainability of SPARK physical education: a case study of translating research into practice. AB - Dissemination and sustainability of evidence-based physical education programs (PE) has been studied rarely. The sustainability of a health-related PE program (SPARK) was independently evaluated in 111 elementary schools in 7 states. Surveys were mailed to schools that had received SPARK curriculum books, training and follow-up (response rate = 47%). Up to 80% of schools that adopted SPARK PE reported sustained use up to 4 years later. Schools using SPARK had more frequent PE classes. Sustained use was related to support from the principal, not previously having a standard PE program, having adequate equipment, and teachers being physically active. Program sustainability was similar in advantaged and disadvantaged schools. Evidence-based PE programs can be sustained up to 4 years. PMID- 15810767 TI - Immediate and delayed bilateral transfer of throwing accuracy in male and female children. AB - In the present study, an attempt was made to examine the nature and persistence of bilateral transfer of a throwing skill for a large sample of male and female children. One hundred sixty children ages 6, 8, 10, and 12 years were randomly assigned to either an experimental or control group with an equal number of boys and girls in each group. The experiment lasted 2 days and consisted of a pretest, a practice phase, an immediate transfer test, and a delayed transfer test. On the pretest, each participant performed 10 trials of a novel one-hand throwing task. Following the pretest, participants in the experimental group practiced the skill with the hand opposite the one used during the pretest until they had successfully reached a designated criterion for their age. Participants in the control group performed a balancing activity. Following the practice phase, all participants performed immediate (10 min later) and delayed (24 hr later) transfer tests under the same conditions as the pretest. The results revealed no group differences on the pretest but significantly higher throwing accuracy for the experimental group than the control group on both transfer tests. In addition, boys' throwing accuracy was significantly superior to the girls. It was concluded that bilateral transfer of throwing accuracy can be both a temporary and relatively persistent phenomenon for children and the superior throwing accuracy for boys is consistent with similar gender differences in throwing distance and throwing velocity (Thomas & French, 1985). PMID- 15810768 TI - Learning to detect error in movement timing using physical and observational practice. AB - Three experiments assessed the possibility that a physical practice participant's ability to render appropriate movement timing estimates may be hindered compared to those who merely observed. Results from these experiments revealed that observers and physical practice participants executed and estimated the overall durations of movement sequences similarly and more accurately than those who were not privy to any previous practice. This was true for a case in which (a) the execution demands for the physical practice participant were relatively high when multiple movement sequences were practiced with a consistent relative time structure but different overall durations (Experiment 1) and (b) the execution demands were relatively modest when only a single sequential motor task was learned (Experiment 2). Moreover, this general set of findings remained true for individuals who had previous experience with physical or observational practice, even when timing estimations were made during tests with no execution demands (Experiment 3). Thus, executing a movement sequence does not appear to interfere with the development of a learner's subjective evaluation of overall timing performance. Specifically, these data provided evidence that recognizing error in movement timing can be accomplished via observation, and, more generally, they add to the growing evidence supporting the claim that observational practice is a legitimate method facilitating the acquisition of sequential movement behaviors. PMID- 15810769 TI - Self-controlled feedback is effective if it is based on the learner's performance. AB - The study follows up on the contention that self-controlled feedback schedules benefit learning because they are more tailored to the performers' needs than externally controlled feedback schedules (Chiviacowsky & Wulf, 2002). Under this assumption, one would expect learning advantages for individuals who decide whether they want to receive feedback after a trial rather than before a trial. Participants practiced a sequential timing task, and all could decide the trials on which they received feedback. One group ("self-after") decided after every trial whether they wanted to receive feedback for that trial while another group ("self-before") made that decision before each trial The self-after group showed learning benefits on a delayed transfer test (novel absolute timing requirements) with regard to overall timing and relative-timing accuracy. Thus, self-controlled feedback was more effective when the learner could make a decision about receiving feedback after the trial. This seems to support the view that self controlled feedback benefits learning, because learners can make a decision about feedback based on their performance on a given trial. PMID- 15810770 TI - The effects of a single reminder trial on retention of a motor skill. AB - Two experiments investigated the effects of a single reminder trial on immediate and delayed retention. Experiment 1 determined if beneficial effects of a reminder trial were a function of task order immediate retention performance benefited only when the reminder trial was practiced in the first block of trials. Experiment 2 added a 24-hr delayed retention test to examine the long term benefits of a reminder trial. Retention performance was enhanced over both delay intervals. The long-term effect extended previous research (Shea & Titzer, 1993) that documented effects after 10 min. The use of a single reminder trial established that intertask comparisons between multiple reminder trials were not a precondition for the reminder trial effect as postulated by Shea and Titzer. PMID- 15810771 TI - Part and whole practice: chunking and online control in the acquisition of a serial motor task. AB - A four-component aiming movement was used to examine the relative effectiveness of part and whole practice. Following a pretest, participants were assigned to one of three practice groups. Participants in a "Whole" group practiced the four components together as a unit. A "No Overlap" group practiced the first two and last two components of the task, alternating every fifth trial. An "Overlap" group practiced the transition between the second and third components on every trial by alternating practice of the first three and last three components every five trials. Participants in all groups improved significantly from pretest to immediate posttest and maintained their performance over a 24-hr delay. Contrary to the "chunking hypothesis," participants in the No Overlap group improved as much as those in the other two groups. Kinematic data indicated that participants in all three groups learned to use response-produced feedback earlier in the individual movement trajectories. Moreover, participants appeared to acquire a general ability to make transitions between movement components rather than specific transitions. The results suggest that segmented or segmented "overlap" practice regimes may benefit learning movement sequences of short duration. PMID- 15810772 TI - Identity tensions in lesbian intercollegiate coaches. AB - Using social identity perspective, we investigated the experiences of 13 lesbian college coaches. Through semistructured interviews, the coaches revealed the daily identity tensions they experienced. There was constant negotiation between their social identities of "coach" and "lesbian." The social context of intercollegiate women's athletics created a complex web that juxtaposed these identities against commanding social norms. The challenges these coaches faced ranged from doing what they believed was best for the well being of their athletes (e.g., fighting homonegativism) versus what was best for their professional well being (e.g., remaining silent). These coaches did not passively accept their fate; while they struggled with identity negotiation, they also found ways to counter the heterosexist atmosphere and create positive social change. PMID- 15810773 TI - Listening to the voices: the experiences of African American female student athletes. AB - "Women of color ... have historically been silenced in society and sport" (Smith, 1992, p. 228). This study examined the sport participation patterns of 12 African American female collegiate student athletes using qualitative methods. Data were collected at a large midwestern university during the 1998-99 academic year. An emergent theme was the effect of silencing by the media, athletic administrators, coaches, and other student athletes on the experiences of African American female student athletes. The findings are presented in the following order: the theoretical framework for the study, an introduction to silencing, an overview of the research analyses, a description of the research setting, and a presentation of the data surrounding the theme of silencing as told through the participants' voices. Following these sections is a discussion and suggestions for future research. PMID- 15810774 TI - Validity and reliability of the 3-day physical activity recall in Singaporean adolescents. PMID- 15810775 TI - Self-controlled observational practice enhances learning. PMID- 15810776 TI - Aviation industry provides incentives to report errors. PMID- 15810777 TI - New tool seeks to measure and improve patient safety. PMID- 15810778 TI - Adverse event reporting: making the system work. PMID- 15810779 TI - The ongoing challenge: catching and keeping quality talent. PMID- 15810780 TI - Part II: Interior troubleshooting. PMID- 15810781 TI - NFPA 99-an overview and discussion of recent changes. PMID- 15810782 TI - Pulse oximeters. PMID- 15810783 TI - An introduction to RFID. PMID- 15810784 TI - Secrets to a successful JCAHO survey. PMID- 15810785 TI - Benefits of equipment swapping within health systems. PMID- 15810786 TI - Flight plan to launch a business. PMID- 15810787 TI - Safety and the gas supply: focus on anesthesia systems. PMID- 15810788 TI - Combination products and the role of medical device firms. PMID- 15810789 TI - Taking a proactive role in adverse event investigations. PMID- 15810790 TI - Enteral feeding monitor/manager. AB - A simple and inherently safe Enteral Feeding Monitor/Manager is described for use with standard, single-lumen jejunal tubes. No feedings or GI fluids are removed permanently unless they accumulate adjacent to the catheter. Only such momentary excess is removed promptly, before local distention can develop. The full bore automatically alternates between 30 seconds on suction and 30 seconds for return of the aspirate. Each minute, the removed air bubbles are vented, the residual jejunal volume is measured, and the degassed fluid is returned by gravity. PMID- 15810791 TI - Power-line interference cancellation in ECG signals. AB - The efficiency of notchfilters and a subtraction procedure for power-line interference cancellation in electrocardiogram (ECG) signals is assessed. In contrast with the subtraction procedure, widely used digital notch filters unacceptably affect QRS complexes. The procedure eliminates interferences of variable amplitude and frequency. The frequency modulations are overcome by adaptive synchronized sampling. Initially, this is accomplished by current hardware power-line frequency measurement. Because this approach is impossible in battery-supplied and some computer-aidd devices, a software measurement of the power-line interference period is developed. PMID- 15810792 TI - Human factors: growing awareness and acceptance. PMID- 15810793 TI - Promote yourself and your profession. PMID- 15810794 TI - Biochemical indicators of nutritional status and dietary intake in Costa Rican Cabecar Indian adolescents. AB - The purpose of this study was to determine the blood levels of selected nutritional status indicators and the dietary intake of Costa Rican Cabecar Indians aged 10 to 16 years. The results showed that 65% of the adolescents had an adequate body mass index (BMI) for their age, and 32% had a BMI < 5th percentile. Likewise, the study revealed a high prevalence of anemia (57%), deficient serum folate levels (54%), deficient vitamin B12 levels (31%), and subclinical vitamin A deficiency (10%). Additionally, the youngsters had elevated prevalences of high triglyceride levels (77%), borderline high-density lipoprotein (HDL) cholesterol levels (46%), homocysteine levels > 10 micromol/L (29%), and homozygous mutation of methylenetetrahydrofolate reductase (MTHFR) (49%). The diet was poor, being high in saturated fat and low in polyunsaturated fat, fiber, and several micronutrients. The dietary intakes of more than 55% of the adolescents did not meet 50% of the estimated average requirements (EAR) for zinc, vitamin A, vitamin C, vitamin B12, vitamin B2, and folate. Furthermore, a high prevalence of parasitosis was found (68%). Our results show an adolescent Cabecar population with a mosaic of nutritional deficiencies and cardiovascular risk factors. PMID- 15810795 TI - Impact of a homestead gardening program on household food security and empowerment of women in Bangladesh. AB - This paper assesses the additional benefits of a homestead gardening program designed to control vitamin A deficiency in Bangladesh. In February and March 2002, data were collected on the food security and social status of women from 2,160 households of active and former participants in the gardening program and from control groups in order to assess the impact and sustainability of the program. The proportions of active and former-participant households that gardened year-round were fivefold and threefold, respectively, higher than that of the control group (78% and 50% vs. 15%). In a three-month period, the households of active participants produced a median of 135 kg and consumed a median of 85 kg of vegetables, while the control households produced a median of 46 kg and consumed a median of 38 kg (p <.001). About 64% of the active participant households generated a median garden income of 347 taka (US$1 = 51 taka), which was spent mainly on food, and 25% of the control households generated 200 taka in the same period (p < .001). The garden production and income levels of formerly participating households three years after withdrawal of program support were much higher than those of the control households, illustrating the sustainability of the program and its ability to increase household food security. Significantly more women in active- and former participant households than in control households perceived that they had increased their economic contribution to their households since the time the program was launched in their subdistricts (> 85% vs. 52%). Similar results were found for the level of influence gained by women on household decision-making. These results highlight the multiple benefits that homestead gardening programs can bring and demonstrate that these benefits should be considered when selecting nutritional and development approaches targeting poor households. PMID- 15810796 TI - Maintaining high vitamin A supplementation coverage in children: lessons from Niger. AB - In 1997, the reduction of child mortality became a policy priority for the Government of Niger because Niger's child mortality rate was the highest in the world. The Ministry of Public Health, Helen Keller International (HKI), and UNICEF spearheaded a coalition-building process linking vitamin A deficiency (VAD) control to national child survival goals. An evidence-based advocacy strategy was developed around the child survival benefits of adequate and sustained VAD control with one unambiguous message: "VAD control can avert over 25,000 child deaths per year." As a result, in 1997 Niger became one of the first countries in Africa to effectively integrate vitamin A supplementation into National Immunization Days (NIDs) for polio eradication. The challenge was then to provide children with a second annual dose of vitamin A. This led in 1999 to the first ever National Micronutrient Days (NMDs) in Africa. NMDs are mobilization campaigns in which caregivers are actively encouraged to take their children for the delivery of vitamin A supplements. Since 1999, the combination of NIDs and NMDs has ensured that over 80% of children 6 to 59 months of age receive two vitamin A doses annually. The success of NIDs/NMDs has relied on five pillars: leadership and ownership by the Ministry of Public Health; district level planning and implementation; effective training and flexible delivery mechanisms; effective social information, communication, and mobilization; and responsiveness and flexibility of Ministry of Public Health and development partners. This successful approach has been widely disseminated, notably through the West African Nutrition Focal Points Network. PMID- 15810797 TI - Milk fortified with iron or iron supplementation to improve nutritional status of pregnant women: an intervention trial from rural Vietnam. AB - Anemia is still the major nutritional problem among pregnant women in Southeast Asia. The objective of this study was to measure hemoglobin status and reduction of underweight in a group of pregnant women who received iron-fortified or nonfortified milk, and another group who received iron supplements (tablets) or placebo. The 44 women in the iron-fortified milk group received 15 mg of iron per day per 400 ml of milk, and 41 women received placebo. The 40 women in the iron supplement group received 60 mg of iron per day, and 43 women received nonfortified milk. During this intervention trial, all women were supervised from the 14th to the 18th week of gestation until delivery. Blood was sampled at 0, 5, 10, and 16 weeks of intervention. After the 16th week of intervention, the changes in hemoglobin (deltaHb) concentrations in both treatment groups (the iron fortified milk and the iron tablet groups) were not significantly different (deltaHb: -0.5+/-0.9 and -0.3+/-0.9 g/L, respectively), but the changes were significantly greater in the nonfortified milk and placebo groups (deltaHb: 1.2+/-0.9 and -1.1+/-0.8 g/L, respectively; p < .01). The change in transferrin saturation (deltaTS) in the iron-fortified milk group (deltaTS: 3.4+/-12.9%) was greater than that in the placebo and nonfortified milk groups (deltaTS: -10.1+/ 9.8% and -11.6+/-10.7 %, respectively) (p < .01). The weight gain of the subjects during intervention did not differ significantly in the fortified and nonfortified milk groups (delta weight: 5.0+/-2.0 and 5.8+/-2.1 kg, respectively), but was higher than in the iron tablet group (delta weight: 4.6+/ 3.1 kg; p < .05) and the placebo group (delta weight: 3.8+/-2.5 kg; p < .001). Iron supplementation and fortification were seen to be effective in promoting weight gain in pregnant Vietnamese women. For women who are underweight, the administration of iron-fortified milk has additional benefits to those of supplementation, most likely due to additional energy and nutrient inputs. PMID- 15810798 TI - Cultural and environmental barriers to adequate iron intake among northern Kenyan schoolchildren. AB - The purpose of this study was to examine the context of iron deficiency and feeding patterns of iron-rich foods among northern Kenyan school-aged children. A nutrition survey was conducted among 300 subjects in two Rendille communities, Korr and Karare. The objectives were to determine the prevalence of iron deficiency as it relates to parasitic infection, dietary intake, and sociodemographic factors, as well as cultural food proscriptions influencing child feeding. Sociodemographic and qualitative data on food beliefs and child feeding practices were obtained from the primary caretaker of each subject. From pediatric subjects, 24-hour dietary recall data were obtained with the help of the primary caretaker, and capillary blood from a fingerstick was used to detect iron deficiency based on measures of hemoglobin, the zinc protoporphyrin-to-heme ratio, C-reactive protein, and transferrin receptor. With an overall prevalence of 31.2%, iron deficiency was found to be associated with dietary iron intakes constrained by diverse economic, cultural, and environmental factors among Rendille children. In Karare, where children's iron intake approached recommended levels, iron deficiency was found to be attributable to low bioavailability of iron (only 4.3% of total iron intake), rather than low dietary intake per se. By contrast, in Korr the average daily iron intake was estimated at only 65% of recommended allowances, indicating that iron deficiency was the outcome not merely of low bioavailability, but rather of overall inadequate iron intake. Sociodemographic analysis showed a significant interaction between sex and economic status, revealing that girls in economically sufficient households were 2.4 times as likely to have iron deficiency as boys. This difference in risk parallels culturally defined gender-based proscriptions for child feeding: girls are believed to benefit from "soft foods," including rice, maize porridge, and tea, whereas boys benefitfrom "hard foods," including meat, blood, and beans. Consequently, in households economically able to purchase iron-rich foods, these foods are being preferentially fed to boys. Economic development may result in improved iron status for boys, but it will be unlikely to benefit girls in the absence of a dietary modification intervention. A modification of culturally acceptable "soft foods" to include iron-rich foods may provide a sustainable approach to controlling and preventing iron deficiency in this population. PMID- 15810800 TI - Recent trends in malnutrition in developing regions: vitamin A deficiency, anemia, iodine deficiency, and child underweight. PMID- 15810799 TI - Prevalence of aflatoxins in blood and urine of Egyptian infants with protein energy malnutrition. AB - The aim of the present work was to study the presence of aflatoxins in blood and urine of infants with protein-energy malnutrition (PEM). The study was conducted on 60 infants, 30 with kwashiorkor and 30 with marasmus, with 10 age-matched healthy infants studied as a control group. Complete blood count, liver function tests, and determination of the level of aflatoxins (B1, B2, G1, G2, M1, M2, G2a, B3, GM1, P, and aflatoxicol R0) in blood and urine were carried out in all studied infants. Serum aflatoxins were detected in more infants with kwashiorkor (80%) than in those with marasmus (46.7%). The mean serum levels of total aflatoxins, AFB1, AFG1, and AFB2a, were significantly higher in infants with kwashiorkor (p <.001). Aflatoxin B1 (AFB1) was the most commonly detected type. The prevalence of aflatoxin excretion in the urine of infants with kwashiorkor was 80%, a higher value than that in infants with marasmus (46.7%). The mean urinary concentration of total aflatoxins followed the same pattern of distribution (p < .052). There were no significant differences between groups in the mean urinary concentrations of AFB1, AFG1, AFB2a, AFM1, and AFG2a. Aflatoxins were not detected in any of the serum or urine samples of the control group. Aflatoxins are highly prevalent in this study population and show a high degree of correlation with severe PEM. PMID- 15810801 TI - Recent trends in malnutrition in developing regions: vitamin A deficiency, anemia, iodine deficiency, and child underweight. PMID- 15810802 TI - Human energy requirements: report of a joint FAO/ WHO/UNU Expert Consultation. PMID- 15810803 TI - Present and future rosacea therapy. AB - Despite its prevalence, rosacea has not received the same attention of researchers as other dermatologic disorders. Nevertheless, new pharmacologic and nonpharmacologic therapies for the condition continue to be developed. The future of rosacea treatment will probably involve a combination of drugs and devices. Certain core therapies (i.e., topical metronidazole, topical azelaic acid, oral tetracyclines, and topical sulfur/sodium sulfacetamide) are validated by the greatest amount of high-order clinical evidence and will undoubtedly remain first line therapeutic choices. However, more research is necessary to validate the efficacy and safety of newer pharmacologic agents and light-based therapy. Because rosacea is a chronic condition, pharmacologic maintenance therapy is necessary to maintain remission. PMID- 15810804 TI - Evidence-based dermatology. AB - Evidence-based dermatology (EBD) is the application of the principles of evidence based medicine to the diagnosis and treatment of skin disorders. EBD does not discount the individual dermatologist's clinical judgment. In fact, EBD is based on the interaction of external evidence, the physician's clinical experience, and the patient's experience. Randomized controlled trials constitute one of the highest levels of evidence and are the gold standard for validating a therapeutic intervention. For the treatment of rosacea, oral tetracycline, topical metronidazole, topical azelaic acid, and topical sulfur/sodium sulfacetamide have been validated by more than one randomized controlled trial. PMID- 15810805 TI - The rigor of trials evaluating Rosacea treatments. AB - The Cochrane Collaboration is an international nonprofit organization that conducts systematic reviews of healthcare interventions. The organization has recently reviewed all studies meeting designated criteria on interventions for rosacea. To be included in the review, trials had to be randomized controlled trials (RCTs) that met the methodological criteria of the reviewers and that were conducted in an adult patient population with moderate to severe rosacea. The electronic databases searched included The Cochrane Skin Group Specialised Trials Register, The Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, BIOSIS, and Science Citation Index. The reviewers tried to obtain details of unpublished and ongoing RCTs through correspondence with authors and pharmaceutical companies. After evaluating the included studies, the reviewers concluded there is evidence that topical metronidazole in 1% cream and 0.75% gel formulations and azelaic acid in 20% cream formulation are effective and safe. Furthermore, there is some evidence that oral metronidazole and tetracycline are effective. The reviewers also made suggestions about future rosacea research. PMID- 15810806 TI - Adjunctive skin care in the management of rosacea: cleansers, moisturizers, and photoprotectants. AB - Certain skin characteristics, such as altered vascular reactivity, appear to be common among patients with rosacea. This may partly explain the observation that these patients appear to have increased sensitivity to certain components of commonly used topical agents. Accordingly, patients with rosacea should be educated regarding which general skin care products to use and to avoid. This review summarizes information regarding 3 classes of these products--cleansers, moisturizers, and photoprotectants--with emphasis on barrier function and skin irritation. PMID- 15810807 TI - Lasers and light sources for rosacea. AB - Pharmacologic agents remain the mainstay for initial and maintenance treatment of rosacea. However, monochromatic (i.e., laser) and polychromatic light-based therapies are increasingly being used for the treatment of certain signs of rosacea. Despite the increased use of lasers and other light-based therapies, few well-controlled studies have been conducted on their use for the treatment of rosacea. The studies that do exist suggest that these modalities have value in treating erythematotelangiectatic rosacea, including persistent erythema and phymatous rosacea. Light-based therapies should be strongly considered in cases of serious erythema, flushing, and telangiectasia because these signs are not optimally addressed by pharmacologic interventions. PMID- 15810808 TI - The pharmacologic therapy of rosacea: a paradigm shift in progress. AB - A number of topical and systemic pharmacologic therapies, some of which remain investigational, have been used to treat rosacea. The pathophysiology of rosacea appears to be inflammatory, and most of the interventions modulate the inflammatory process in some way. Topical agents include various formulations of sodium sulfacetamide and sulfur, metronidazole, azelaic acid, and benzoyl peroxide/clindamycin. Oral agents include antibiotics in conventional and subantimicrobial doses. A paradigm shift in progress in the management of rosacea encompasses the use of these and other agents either alone or, increasingly, in different combinations, based on the subtype of rosacea. PMID- 15810809 TI - Cancer vaccines 2004 opening address: the molecular and cellular basis of cancer immunosurveillance and immunoediting. PMID- 15810810 TI - Organic field-effect transistors from solution-deposited functionalized acenes with mobilities as high as 1 cm2/V x s. AB - We present the device parameters for organic field-effect transistors fabricated from solution-deposited films of functionalized pentacene and anthradithiophenes. These materials are easily prepared in one or two steps from commercially available starting materials and are purified by simple recrystallization. For a solution-deposited film of functionalized pentacene, hole mobility of 0.17 cm2/V.s was measured. The functionalized anthradithiophenes showed behavior strongly dependent on the substituents, with hole mobilities as high as 1.0 cm2/V.s. PMID- 15810811 TI - Demonstration of the elusive concentration-gradient paramagnetic force. AB - Using classical electrochemistry, it is demonstrated that the concentration gradient paramagnetic force, FnablaC, is a body force proportional to |B|2 acting parallel to the concentration gradient of electrogenerated radicals. FnablaC can balance gravity, holding volumes of solution wherein mass transfer continues to take place by diffusion. In contrast to usual levitation forces, FnablaC does not depend on field gradients and may be present even in homogeneous magnetic fields. Understanding the properties of FnablaC is relevant to magnetic confinement and levitation and is speculated even to propulsion with objects having permanent susceptibility gradients. PMID- 15810812 TI - Silica-coated nanocomposites of magnetic nanoparticles and quantum dots. AB - Quantum dots (QDs) and magnetic nanoparticles (MPs) are of interest for biological imaging, drug targeting, and bioconjugation because of their unique optoelectronic and magnetic properties, respectively. To provide for water solubility and biocompatibility, QDs and MPs were encapsulated within a silica shell using a reverse microemulsion synthesis. The resulting SiO2/MP-QD nanocomposite particles present a unique combination of magnetic and optical properties. Their nonporous silica shell allows them to be surface modified for bioconjugation in various biomedical applications. PMID- 15810813 TI - IR study of cross-strand coupling in a beta-hairpin peptide using isotopic labels. AB - Model beta-hairpin peptides can be used to develop understanding of fundamental elements of beta-sheet secondary structure formation and stability. We have studied two 13C-labeled variants of a beta-hairpin peptide modified from a design originally proposed by Gellman: Arg-Tyr-Val-Glu-Val-Aib-Gly-Lys-Lys-Ile-Leu-Gln. (In this peptide, the two italicized residues form a beta-turn, while 13C-labels are on the amide C=O of Val3, Lys8 in HBG-L and Val3, Ile10 in HBG-S.) Both these peptides are labeled on opposite strands of the hairpin, but differ in the labeling pattern. One (HBG-L) forms a large (14-atom) H-bonded ring of labeled C=Os, while the other (HBG-S) forms a small (10-atom) H-bonded ring. These impact the amide I infrared spectra, with HBG-L having a 13C frequency and intensity higher than that of HBG-S, in good agreement with our spectral simulations based on quantum mechanically derived force fields. The thermal behavior of both peptides yields a broad thermal transition and lacks an isosbestic point. The 13C band for HBG-L has the largest intensity change with temperature, distinct from the 12C change and the HBG-S 13C change. PMID- 15810814 TI - Detection of intermediates in cobalt-catalyzed hydroformylation using para hydrogen-induced polarization. AB - para-Hydrogen-induced polarization methods are shown to enable the in situ detection of linear and branched monophosphine-containing intermediates during hydroformylation when Co(eta3-C3H5)(CO)2(PCy3) is the catalyst precursor. The NMR signal characteristics of the alkyl arms of these species provide direct evidence for the rapid interconversion of linear and branched cobalt alkyls prior to the CO insertion step. The observation of additional para-hydrogen-enhanced signals for the corresponding linear and branched aldehydes enables the reactions selectivity to be rapidly monitored as a function of H2 and CO pressure or reaction temperature. PMID- 15810815 TI - Direct C-arylation of free (NH)-indoles and pyrroles catalyzed by Ar-Rh(III) complexes assembled in situ. AB - Ar-Rh(III) pivalate complexes assembled in situ from the reaction of [RhCl(coe)2]2 (coe = cis-cyclooctene), [p-(CF3)C6H4]3P, and CsOPiv effectively catalyzed the direct C-arylation of free (NH)-indoles and (NH)-pyrroles in good yields and with high regioselectivity. The reaction displayed excellent functional group compatibility and low moisture sensitivity. Kinetics studies support a mechanism involving phosphine displacement by indole in complex 2 (resting state of the catalyst), followed by a rate-limiting C-H bond metalation. PMID- 15810816 TI - Selective formation of MSi16 (M = Sc, Ti, and V). AB - Metal-encapsulated silicon cage clusters are a new class of clusters and are opening up new avenues for silicon-based nanoscale materials. We present experimental evidence for a highly stable cluster corresponding to M@Si16 (M = Sc, Ti, and V). Mass spectrometry and anion photoelectron spectroscopy show that the cluster features an electronically closed TiSi16 neutral core which undergoes a change in the number of valence electrons involving (i) substitution of neighboring metals with Sc and V, or (ii) addition of a halogen atom to the TiSi16 anion, and that VSi16F is predicted to form an ionically bound superatom complex. PMID- 15810817 TI - Protein biosensors based on biofunctionalized conical gold nanotubes. AB - There is increasing interest in the concept of using nanopores as the sensing elements in biosensors. The nanopore most often used is the alpha-hemolysin protein channel, and the sensor consists of a single channel embedded within a lipid bilayer membrane. An ionic current is passed through the channel, and analyte species are detected as transient blocks in this current associated with translocation of the analyte through the channel-stochastic sensing. While this is an extremely promising sensing paradigm, it would be advantageous to eliminate the very fragile lipid bilayer membrane and perhaps to replace the biological nanopore with an abiotic equivalent. We describe here a new family of protein biosensors that are based on conically shaped gold nanotubes embedded within a mechanical and chemically robust polymeric membrane. While these sensors also function by passing an ion current through the nanotube, the sensing paradigm is different from the previous devices in that a transient change in the current is not observed. Instead, the protein analyte binds to a biochemical molecular recognition agent at the mouth of the conical nanotube, resulting in complete blockage of the ion current. Three different molecular-recognition agents, and correspondingly three different protein analytes, were investigated: (i) biotin/streptavidin, (ii) protein-G/immunoglobulin, and (iii) an antibody to the protein ricin with ricin as the analyte. PMID- 15810818 TI - Pairwise coupling in an Arg-Phe-Met triplet stabilizes alpha-helical peptide via shared rotamer preferences. AB - The hydrophobic Arg-Phe and Phe-Met side chain interactions stabilize the alpha helix by -0.29 and -0.59 kcal/mol, respectively, when placed i, i + 4 in an alanine-based peptide. When both interactions are present simultaneously, however, they stabilize the helix by an additional -0.75 kcal/mol, nearly as much as the sum of its parts. We attribute this coupling to a shared rotamer preference, as the central Phe is t in both bonds. The energetic cost of restricting the Phe residue into a t conformation is only paid once in the triplet, rather than twice when the interactions are separate. Coupling is thus demonstrated to have large effects on protein stability. PMID- 15810819 TI - Convergent synthesis of a fully lipidated glycosylphosphatidylinositol anchor of Plasmodium falciparum. AB - A highly convergent strategy for the synthesis of fully lipidated GPI anchors of malarial origin is reported. This strategy utilized three orthogonal protecting groups, which can be chemoselectively deprotected and functionalized in the late stage of the synthesis. Rapid access to the target GPIs in a highly efficient manner in sufficient quantities for the biological studies has been achieved. PMID- 15810820 TI - Vesicle formed by amphiphilc cucurbit[6]uril: versatile, noncovalent modification of the vesicle surface, and multivalent binding of sugar-decorated vesicles to lectin. AB - We report a novel vesicle formed by an amphiphilic CB[6] derivative, the surface of which can be easily modified via host-guest interactions by taking advantage of molecular cavities, readily accessible at the vesicle surface, and their strong affinity toward polyamines. Amphiphilic CB[6] derivative 1 synthesized by reaction between (allyloxy)12CB[6] and 2-[2-(2-methoxyethoxy)ethoxy]ethanethiol affords a vesicle that has been characterized by TEM, light scattering, and fluorescent dye entrapment experiments. Treatment of vesicle 1 with FITC (fluorescein isothiocyanate)-spermine conjugate ligand 2, in which spermine serves as a binding motif to CB[6] and FITC as a fluorescent tag, produced a surface-modified vesicle, which can be easily visualized by a confocal microscope. This result provides us with a new noncovalent, modular approach to the modification of vesicle surfaces. By treating the vesicle derived from the amphiphilic CB[6] with a tag-attached polyamine, we can easily decorate the surface of the vesicle with the tag. Sugar-decorated vesicles were prepared by this noncovalent method, and their interactions with concanavalin A (ConA) were studied. The binding constant of the vesicle decorated with mannose-spermidine conjugate 3 to ConA was measured to be approximately 3 x 104 M-1, which is almost 3 orders of magnitude higher than that of free ligand 3 to ConA (K = approximately 50 M-1). On the other hand, the binding constant of the vesicle coated with galactose-spermidine conjugate 4 to ConA was too small to be measured. These results illustrate the specific and multivalent interactions between the mannose-decorated vesicle and ConA. The ability for facile surface modification suggests many practical applications, including its use in targeted drug delivery and immunization. PMID- 15810821 TI - Structural and magnetic evidence concerning spin crossover in formamidinate compounds with ru2 5+ cores. AB - The magnetic and structural properties of two Ru2(DArF)4Cl compounds, where DArF is the anion of a diaryl formamidine, are presented here. The compounds with Ar = p-anisyl and m-anisyl both show temperature dependence of chiT (chi = molar magnetic susceptibility), but for different reasons. For the para compound, there is a Boltzmann distribution between a pi*3 ground state and a delta*pi*2 upper state, and this is confirmed by a temperature dependence of the Ru-Ru bond length: 2.4471(5) A at 23 K and 2.3968(5) A at 300 K. For the meta compound, a delta*pi*2 configuration persists over the range of 23-300 K as shown by an invariant Ru-Ru bond length, but the chiT drops with decreasing temperature owing to zero-field splitting of a 4B2u ground state. PMID- 15810822 TI - Room-temperature ferroelectricity and gigantic dielectric susceptibility on a supramolecular architecture of phenazine and deuterated chloranilic acid. AB - Ferroelectricity and large dielectric constant at room temperature have been demonstrated for cocrystals of phenazine and anilic acids constructing supramolecular assemblies. Deuteration of the anilic acids gives rise to an increase by more than 50 K in the transition temperature, which exceeds room temperature for the deuterated chloranilic acid. At room temperature in air, the crystals show a clear polarization hysteresis with a small coercive field. Application of hydrostatic pressure and halogen substitution in the anilic acids bring about the structural effect on the transition temperature in a parallel way, whereas the deuterium substitution does this in a distinct way. The observed large deuteration effect cannot be elucidated by the geometric change of the hydrogen bond, which has been considered as the possible mechanism of ferroelectricity in the conventional hydrogen-bonded with double-well potential. PMID- 15810823 TI - Mixed-metal triangular trinuclear complexes: dimers of gold-silver mixed-metal complexes from gold(I) carbeniates and silver(I) 3,5-diphenylpyrazolates. AB - Dimers of trinuclear mixed gold-silver compounds are obtained by the reaction of a gold(I) carbeniate, [Au(mu-C(OEt)=NC6H4-p-CH3)]3, with a silver(I) pyrazolate, [Ag(mu-3,5-Ph2pz)]3. The crystalline products are the mixed-metal species Au(carb)Ag2(mu-3,5-Ph2pz)2 and Au2(carb)2Ag(mu-3,5-Ph2pz)CCH2Cl2. PMID- 15810824 TI - Decomposition of ammonia and hydrogen on Ir surfaces: structure sensitivity and nanometer-scale size effects. AB - The adsorption and decomposition of ammonia and hydrogen have been studied on surfaces of clean planar Ir(210) and clean nanoscale-faceted Ir(210), which are prepared from the same crystal in situ. We find evidence for structure sensitivity in recombination and desorption of H2 and in thermal decomposition of NH3 on clean planar Ir(210) versus clean faceted Ir(210). Moreover, the decomposition kinetics of NH3 on faceted Ir(210) exhibit size effects on the nanometer scale, which is the first observation of size effects in surface chemistry on an unsupported monometallic catalyst with controlled and well defined structure and size. PMID- 15810825 TI - Chiral attachment of styrene mediated by surface dimers on Ge100. AB - We have investigated the chiral adsorption configurations of styrene on Ge(100) using scanning tunneling microscopy at 300 K. The chemisorbed styrene on a single Ge dimer reduces the symmetry of the molecule, which produces a chiral center, and leads to the (S) or (R) chiral on-top configuration. We have found that the dimeric adsorption of styrene induced by the Ge surface dimer structure forms the enantiomeric and diastereomeric paired end-bridge configurations. We determine the absolute chirality of adsorbed styrene on Ge(100) and demonstrate a novel method for the achiral molecule to produce dimeric enantiomers and diastereomers attached to the semiconductor surface. PMID- 15810826 TI - Dual memory of enantiomeric helices in a polyacetylene induced by a single enantiomer. AB - We report the dual memory of both the enantiomeric right- and left-handed helical conformations induced in a polyacetylene based on the temperature-stimulated helicity inversion of the polymer. The polyacetylene folds into a one-handed helix induced by noncovalent bonding interactions with a single enantiomeric amine. The induced helix underwent a reversible inversion of the helicity by temperature. The diastereomeric right- and left-handed helices obtained at different temperatures could be further memorized when the optically active amine was replaced by an achiral diamine, generating right- and left-handed helices of the mirror images of each other. Consequently, both enantiomeric helices can be produced with a high efficiency from dynamically diastereomeric helical polyacetylenes induced by a single enantiomer. PMID- 15810827 TI - Enhanced para-xylene selectivity in the toluene alkylation reaction at ultralow contact time. AB - Dramatic improvements in the para-xylene selectivity of the toluene alkylation reaction can be effected by operating the catalytic reaction at ultralow contact time. Unexpectedly, the rate of alkylation is sustained, while unwanted side reactions are suppressed. By demonstrating that contact time directly influences the fate of para-xylene, which is known to form and diffuse preferentially within the zeolite catalyst, we conclude that external mass transfer is a key parameter in controlling selectivity. Even non-optimized catalysts can be made to achieve near-perfect selectivity, without sacrificing conversion. PMID- 15810828 TI - Intermolecular and intramolecular, platinum-catalyzed, acceptorless dehydrogenative coupling of hydrosilanes with aryl and aliphatic methyl C-H bonds. AB - Intermolecular acceptorless dehydrogenative coupling of silanes with arene C-H bonds and intramolecular coupling of silanes with aryl and alkyl C-H bonds occur in good yield in the presence of 5 mol % of TpMe2PtMe2H (TpMe2 = hydridotris(3,5 dimethylpyrazolyl)borate) and related platinum(IV) complexes. The intermolecular reactions of arenes occurred with both trialkyl and dialkylaryl silanes. Intramolecular reactions of dialkylsilylalkylarenes occurred at aryl C-H bonds, and reactions of tributylsilane or dibutylphenylsilane occurred intramolecularly at the aliphatic, primary C-H bond. The reactions of arenes occurred preferentially at the least sterically hindered C-H bonds and preferentially with more electron-poor arenes. Crossover experiments and the lack of reactivity of the arylsilanes with H2 imply that the dehydrogenative silylation of arenes can be irreversible, even in a closed reaction vessel. PMID- 15810829 TI - UV-Vis microspectroscopy: probing the initial stages of supported metal oxide catalyst preparation. AB - A UV-vis microspectroscopy methodology for monitoring the speciation and macrodistribution of catalyst-precursor species inside catalyst-support bodies at the initial stages of catalyst preparation has been developed. The setup is based upon optical-fiber technology and allows spatially resolved analysis of bisected catalyst bodies. The potential of this tool is demonstrated by two pore-volume impregnation studies involving Ni2+ d-d transition bands and Cr6+ charge-transfer bands. PMID- 15810831 TI - A tandem carbanion addition/carbon-carbon bond cleavage yields alkynyl ketones. AB - Carbanion addition to vinylogous acid triflates triggers carbon-carbon bond cleavage to form alkynyl ketones under mild conditions. Mechanistic factors affecting the cleavage event and its relationship to complementary fragmentations are discussed. A range of tethered keto-alkynes are obtained by a unified approach. PMID- 15810830 TI - Nucleobase participation in ribozyme catalysis. AB - We constructed a modified form of the VS ribozyme containing an imidazole ring in place of adenine at position 756. The novel ribozyme is active in both cleavage and ligation reactions. The reaction is efficient, although relatively slow. The results are consistent with a role for nucleobase catalysis in the catalytic mechanism of this ribozyme. PMID- 15810832 TI - A nickel-catalyzed route to pyridines. AB - A mild and general route for preparing pyridines from nitriles and diynes is described. Ni/imidazolyidene complexes were used to mediate cyclization alkynes and both aryl and alkyl nitriles at ambient temperature. In addition, the efficacy of this protocol allows for the preparation of a fused seven-membered pyridone and for intermolecular cyclizations. When an asymmetrical diyne was employed, cyclization afforded a single pyridine regioisomer. PMID- 15810833 TI - Direct observation of a 14-electron ruthenacyclobutane relevant to olefin metathesis. AB - The 14-electron ruthenium phosphonium alkylidene complex [(IH2Mes)Cl2Ru=CH(PCy3)][B(C6F5)4], 1b, a highly active olefin metathesis catalyst, reacts with stoichiometric quantities of ethylene at -50 degrees C in CD2Cl2 to generate the ruthenacyclobutane complex [(IH2Mes)Cl2RuCH2CH2CH2], 2, and [CH2=CH(PCy3)][B(C6F5)4] in quantitative yield by NMR spectroscopy. 1H and 13C NMR spectroscopies on 2 and 2-13C3 are consistent with a symmetrical C2v structure, providing the first experimental information concerning this crucial intermediate in ruthenium-mediated olefin metathesis. At -50 degrees C, exchange with free ethylene takes place on the chemical time scale. Complex 2 decomposes in solution upon warming to room temperature, generating propene and unknown ruthenium product(s). PMID- 15810834 TI - Fast, comprehensive two-dimensional HPLC separation of tryptic peptides based on high-temperature HPLC. AB - Two-dimensional HPLC (2D-LC) has recently received considerable attention, and is being used as an alternative to 2D gel electrophoresis in proteomics research. The greatest impediment to the widespread use of 2D-LC is the long analysis time ranging up to days per analysis, making the technique impractical for many jobs. Here we focus on improving the speed of gradient separations since these are typically used as the second dimension in peptide separations by 2D-LC. Specifically we describe high-temperature, ultrafast HPLC conditions, along with the instrument modifications needed to reduce the analysis time of each complete second-dimension gradient separation to tens of seconds. Most importantly, this system is capable of generating a high peak capacity (1350) characteristic of comprehensive 2D-LC in a relatively shorter analysis time (20 min) with a sampling rate sufficient to minimize information loss with simpler instrumentation than currently used; this is equivalent to one unit of peak capacity per second. PMID- 15810835 TI - Control of nanoparticle location in block copolymers. AB - A simple procedure is described to incorporate gold nanoparticles and control their location within symmetric poly(styrene-b-2 vinyl pyridine) (PS-PVP) diblock copolymers. Gold nanoparticles coated with thiol-terminated PS and/or PVP homopolymer chains (Mn approximately 1300 and 1500 g/mol, respectively) are incorporated into alternating lamellar layers of PS and PVP (total Mn approximately 196 500 g/mol). The location of the particles is controlled by varying the composition of ligands on the particle surfaces. In particular, gold particles coated with 100% PS or PVP reside near the center of the respective polymer domains, while particles coated with a mixture of both homopolymers reside at the interfaces between the two blocks. PMID- 15810836 TI - Metal-templated diyne cyclodimerization and cyclotrimerization. AB - Reaction of [Pt(CH3)2(COD)] (COD = 1,5-cyclooctadiene) with Ph2PCCCCPPh2 led to a mixture of [{Pt(CH3)2}2(mu-Ph2PC4PPh2)2] (1) and [{Pt(CH3)2}3(mu-Ph2PC4PPh2)3] (2). Reaction of [PtCl2(COD)] with Ph2PCCCCPPh2 led to a mixture of the thermally unstable compounds [{PtCl2}2(mu-Ph2PC4PPh2)2] (3) and [{PtCl2}3(mu-Ph2PC4PPh2)3] (4) which transform into [{PtMe2}2{mu-C8(PPh2)4}] (5) and [{PtMe2}3{mu3 C12(PPh2)6}] (6) containing 8-membered diene-diyne and 12-membered triene-triyne rings, respectively. Compound 2 can be converted to [{PtMe2}3{C12(PPh2)6}] (7) by heating with CuCl at 80 degrees C, while 1 can be heated without significant cycloaddition. PMID- 15810837 TI - Highly active, regioselective, and enantioselective hydroformylation with Rh catalysts ligated by Bis-3,4-diazaphospholanes. AB - Azines made by the reaction of hydrazine with ortho-formylbenzoic acid react with 1,2-diphosphinobenzene and either succinyl chloride or phthaloyl chloride in ca. 30% yield to give rac-bis-3,4-diazaphospholanes bearing benzoic acid groups in the 2 and 5 positions. Condensation of the benzoic acid functionalities with enantiomerically pure amines affords diastereomeric benzoamides which can be separated by flash chromatography. Application of the resolved bis-3,4 diazaphosholanes to Rh-catalyzed enantioselective hydroformylation of styrene, allyl cyanide, and vinyl acetate under mild pressures (20-500 psig of CO/H2) and temperatures (40-120 degrees C) reveals high activities and selectivities for all three substrates. At 60 degrees C and 500 psig syn gas, the best ligand provides outstanding regio- and enantioselectivities (styrene, 89% ee, b:l = 30:1; allyl cyanide, 87% ee, b:l = 4.8:1; vinyl acetate, 95% ee, b:l = 40:1) while achieving turnover frequencies of ca. 3000 h-1. PMID- 15810838 TI - Localized surface plasmon resonance sensing of lipid-membrane-mediated biorecognition events. AB - Supported phospholipid bilayers (SPBs) have emerged as important model systems for studies of the natural cell membrane and its components, which are essential for the integrity and function of cells in all living organisms, and also constitute common targets for therapeutic drugs and in disease diagnosis. However, the preferential occurrence of spontaneous SPB formation on silicon based substrates, but not on bare noble-metal surfaces, has so far excluded the use of the localized surface plasmon resonance (LSPR) sensing principle for studies of lipid-membrane-mediated biorecognition reactions. This is because the LSPR phenomenon is associated with, and strongly confined to, the interfacial region of nanometric noble-metal particles. This problem has been overcome in this study by a self-assembly process utilizing localized rupture of phospholipid vesicles on silicon dioxide in the bottom of nanometric holes in a thin gold film. The hole-induced localization of the LSPR field to the voids of the holes is demonstrated to provide an extension of the LSPR sensing concept to studies of reactions confined exclusively to SPB-patches supported on SiO2. In particular, we emphasize the possibility of performing label-free studies of lipid-membrane mediated reaction kinetics, including the compatibility of the assay with array based reading (approximately 7 x 7 microm2) and detection of signals originating from bound protein in the zeptomole regime. PMID- 15810839 TI - Role of protonation and of axial ligands in the reductive dechlorination of alkyl chlorides by vitamin B12 complexes. Reductive cleavage of chloroacetonitrile by Co(I) cobalamins and cobinamides. AB - Cobalt(I) cobalamin and cobinamide are efficient catalysts of the hydrogenolysis of aliphatic chloro compounds. Taking chloroacetonitrile as example, the first requirement for high catalytic efficiency is fulfilled by the high reactivity of the Co(I) complex toward the substrate, leading to the alkylcobalt(III) complex. This is further reduced into the alkylcobalt(II) complex. However, the fact that these two reactions are fast is not enough to ensure an efficient catalysis: in DMF catalysis is very poor, while it is high in water. The experiments carried out in DMF with addition of an acid show that a crucial step in the catalytic process is the proton transfer decomposition of the alkylcobalt(II) complex, leading to the product, and closing the catalytic loop by regeneration of the cobalt(I) complex. Another important feature of these catalytic reactions is the role played by axial ligands present in the solution, particularly those that are produced by the catalytic reaction itself, namely, chloride ions and the counteranion of the added acid. The stronger these ligands, the more negative the potential required for the reduction of the alkylcobalt(III) complex. This amounts to a self-moderation effect: the more efficient catalysis, the slower its second step, i.e., the conversion of the alkylcobalt(III) complex into the alkylcobalt(II). PMID- 15810840 TI - Deoxythreosyl phosphonate nucleosides as selective anti-HIV agents. AB - Out of a series of eight new phosphonate nucleosides with an l-threose and an l-2 deoxythreose sugar moiety, two new compounds were identified (PMDTA and PMDTT) that showed potent anti-HIV-1 (HIV-2) activity [EC50 = 2.53 microM (PMDTA) and 6.59 microM (PMDTT)], while no cytoxicity was observed at the highest concentration tested [CC50 > 316 microM (PMDTA) and > 343 microM (PMDTT)]. The kinetics of incorporation of PMDTA into DNA (using the diphosphate of PMDTA as substrate and HIV-1 reverse transcriptase as catalyst) was similar to the kinetics observed for dATP, while the diphosphate of PMDTA was a very poor substrate for DNA polymerase alpha. The incorporated PMDTA fits very well in the active site pocket of HIV-1 reverse transcriptase. PMID- 15810841 TI - An 15N NMR spin relaxation dispersion study of the folding of a pair of engineered mutants of apocytochrome b562. AB - 15N relaxation dispersion NMR spectroscopy has been used to study exchange dynamics in a pair of mutants of Rd-apocyt b562, a redesigned four-helix-bundle protein. An analysis of the relaxation data over a range of temperatures establishes that exchange in both proteins is best modeled as two-state and that it derives from the folding/unfolding transition. These results are in accord with predictions based on the reaction coordinate for the folding of the protein determined from native-state hydrogen exchange data [Chu, R.; Pei, W.; Takei, J.; Bai, Y. Biochemistry 2002, 41, 7998-8003]. The kinetics and thermodynamics of the folding transition have been characterized in detail. Although only a narrow range of temperatures could be examined, it is clear that the folding rate temperature profile is distinctly non-Arrhenius for both mutants, with the folding barrier for at least one of them entropic. PMID- 15810842 TI - Highly enantioselective phase-transfer-catalyzed alkylation of protected alpha amino acid amides toward practical asymmetric synthesis of vicinal diamines, alpha-amino ketones, and alpha-amino alcohols. AB - Highly enantioselective alkylation of protected glycine diphenylmethyl (Dpm) amide 1 and Weinreb amide 10 has been realized under phase-transfer conditions by the successful utilization of designer chiral quaternary ammonium salts of type 4 as catalyst. Particularly, remarkable reactivity of the chiral ammonium enolate derived from 1b and 4c allowed the reaction with less reactive simple secondary alkyl halides with high efficiency and enantioselectivity. An additional unique feature of this chiral ammonium enolate is its ability to recognize the chirality of beta-branched primary alkyl halides, which provides impressive levels of kinetic resolution and double stereodifferentiation during the alkylation, allowing for two alpha- and gamma-stereocenters to be controlled. Combined with the subsequent reduction using LiAlH4 in cyclopentyl methyl ether (CPME), this system offers a facile access to structurally diverse optically active vicinal diamines. Furthermore, the optically active alpha-amino acid Weinreb amide 11 can be efficiently converted to the corresponding amino ketone by a simple treatment with Grignard reagents. In addition, reduction and alkylation of the optically active alpha-amino ketone into both syn and anti alpha-amino alcohols with almost complete relative and absolute stereochemical control have been achieved. With (S,S)- and (R,R)-4 in hand, the present approach renders both enantiomers of alpha-amino amides including Weinreb amides readily available with enormous structural variation and also establishes a general and practical route to vicinal diamines, alpha-amino ketones, and alpha-amino alcohols with the desired stereochemistry. PMID- 15810843 TI - Di- and trivalent organogermanium reactive intermediates. Kinetics and mechanisms of some reactions of diphenylgermylene and tetraphenyldigermene in solution. AB - The reactivity of diphenylgermylene (Ph2Ge) with several classes of germylene scavengers has been studied in hexane solution at 23 degrees C by laser flash photolysis of 3,4-dimethyl-1,1-diphenyl-1-germacyclopent-3-ene (1a), a clean and highly efficient precursor to the germylene and its dimer, tetraphenyldigermene (2a). The reactions studied include M-H insertion reactions with Group 14 hydrides (M = Si, Ge, Sn), halogen atom abstractions from bromo- and chlorocarbons, Lewis acid-base complexation with 1 degrees, 2 degrees, and 3 degrees aliphatic amines, and reaction with an aliphatic alkene, alkyne and diene, and oxygen. Absolute rate constants for (irreversible) scavenging of the germylene could be obtained by direct measurement of the germylene decay kinetics for all but the least efficient scavengers (triethylsilane, oxygen, chloroform, and 1-bromopentane), for which estimates of the rate constants were obtained by Stern-Volmer analysis of the reduction in digermene yield as a function of scavenger concentration. Distinctly different kinetic behavior is observed for scavenging of Ph2Ge by isoprene, 4,4-dimethyl-1-pentene, and triethylamine; in these cases, the results suggest that reaction is rapid (k(Q) = 3-6 x 10(9) M( 1)s(-1)) but reversible (K(eq) = 2500 - ca. 20,000 M(-1)) over the range of scavenger concentrations studied. The reactions with the C-C unsaturated compounds proceed via the intermediacy of long-lived transient species absorbing at <290 nm, which are tentatively assigned to the corresponding three-membered germanocycles on the basis of their UV spectra and lifetimes. Upper limits for the absolute rate constants for reaction of tetraphenyldigermene (2a) toward many of these reagents are also reported. PMID- 15810844 TI - Dendrimer-encapsulated Pd nanoparticles as aqueous, room-temperature catalysts for the Stille reaction. AB - We report that dendrimer-encapsulated Pd nanoparticles having a diameter of approximately 1.7 nm are effective and general catalysts for coupling aryl halides to organostannanes (the Stille reaction) under mild conditions. The significant results of this study are that the Stille reaction is catalyzed by dendrimer-encapsulated Pd nanoparticles in very good yield, in aqueous solution at 23 degrees C, and using only 0.100 atom % of Pd as catalyst. PMID- 15810845 TI - New insights into the interactions of serum proteins with bis(maltolato)oxovanadium(IV): transport and biotransformation of insulin enhancing vanadium pharmaceuticals. AB - Significant new insights into the interactions of the potent insulin-enhancing compound bis(maltolato)oxovanadium(IV) (BMOV) with the serum proteins, apo transferrin and albumin, are presented. Identical reaction products are observed by electron paramagnetic resonance (EPR) with either BMOV or vanadyl sulfate (VOSO4) in solutions of human serum apo-transferrin. Further detailed study rules out the presence of a ternary ligand-vanadyl-transferrin complex proposed previously. By contrast, differences in reaction products are observed for the interactions of BMOV and VOSO4 with human serum albumin (HSA), wherein adduct formation between albumin and BMOV is detected. In BMOV-albumin solutions, vanadyl ions are bound in a unique manner not observed in comparable solutions of VOSO4 and albumin. Presentation of chelated vanadyl ions precludes binding at the numerous nonspecific sites and produces a unique EPR spectrum which is assigned to a BMOV-HSA adduct. The adduct species cannot be produced, however, from a solution of VOSO4 and HSA titrated with maltol. Addition of maltol to a VOSO4-HSA solution instead results in formation of a different end product which has been assigned as a ternary complex, VO(ma)(HSA). Furthermore, analysis of solution equilibria using a model system of BMOV with 1-methylimidazole (formation constant log K1 = 4.5(1), by difference electronic absorption spectroscopy) lends support to an adduct binding mode (VO(ma)2-HSA) proposed herein for BMOV and HSA. This detailed report of an in vitro reactivity difference between VOSO4 and BMOV may have bearing on the form of active vanadium metabolites delivered to target tissues. Albumin binding of vanadium chelates is seen to have a potentially dramatic effect on pharmacokinetics, transport, and efficacy of these antidiabetic chelates. PMID- 15810846 TI - Poisoning of heterogeneous, late transition metal dehydrocoupling catalysts by boranes and other group 13 hydrides. AB - Borane reagents are widely used as reductants for the generation of colloidal metals. When treated with a variety of heterogeneous catalysts such as colloidal Rh, Rh/Al2O3, and Rh(0) black, BH3.THF (THF = tetrahydrofuran) was found to generate H2 gas with the concomitant formation of a passivating boron layer on the surface of the Rh metal, thereby acting as a poison and rendering the catalyst inactive toward the dehydrocoupling of Me2NH.BH3. Analogous poisoning effects were also detected for (i) colloidal Rh treated with other species containing B-H bonds such as [HB-NH]3, or Ga-H bonds such as those present in GaH3.OEt2, (ii) colloidal Rh that was generated from Rh(I) and Rh(III) salts using borane or borohydrides as reductants, and (iii) for other metals such as Ru and Pd. In contrast, analogous poisoning effects were not detected for the catalytic hydrogenation of cyclohexene using Rh/Al2O3 or the Pd-catalyzed Suzuki cross-coupling of PhB(OH)2 and PhI. These results suggest that although this poisoning behavior is not a universal phenomenon, the observation that such boron layers are formed and surface passivation may exist needs to be carefully considered when borane reagents are used for the generation of metal colloids for catalytic or materials science applications. PMID- 15810847 TI - Effect of SOCl2 treatment on electrical and mechanical properties of single-wall carbon nanotube networks. AB - Chemical modification by SOCl2 of an entangled network of purified single-wall carbon nanotubes, also known as 'bucky paper', is reported to profoundly change the electrical and mechanical properties of this system. Four-probe measurements indicate a conductivity increase by up to a factor of 5 at room temperature and an even more pronounced increase at lower temperatures. This chemical modification also improves the mechanical properties of SWNT networks. Whereas the pristine sample shows an overall semiconducting character, the modified material behaves as a metal. The effect of SOCl2 is studied in terms of chemical doping of the nanotube network. We identified the microscopic origin of these changes using SEM, XPS, NEXAFS, EDX, and Raman spectroscopy measurements and ab initio calculations. We interpret the SOCl2-induced conductivity increase by p type doping of the pristine material. This conclusion is reached by electronic structure calculations, which indicate a Fermi level shift into the valence band, and is consistent with the temperature dependence of the thermopower. PMID- 15810848 TI - Reactions of vinyl acetate and vinyl trifluoroacetate with cationic diimine Pd(II) and Ni(II) alkyl complexes: identification of problems connected with copolymerizations of these monomers with ethylene. AB - Vinyl acetate (VA) and vinyl trifluoroacetate (VA(f)) react with [(NwedgeN)Pd(Me)(L)][X] (M = Pd, Ni, (NwedgeN) = N,N'-1,2-acenaphthylenediylidene bis(2,6-dimethyl aniline), Ar(f) = 3,5-trifluoromethyl phenyl, L = Ar(f)CN, Et2O; X = B(Ar(f))4-, SbF6-) to form pi-adducts 3 and 5 at -40 degrees C. Binding affinities relative to ethylene have been determined. Migratory insertion occurs in a 2,1 fashion (DeltaG++ = 19.4 kcal/mol, 0 degrees C for VA, and 17.4 kcal/mol, -40 degrees C for VA(f)) to yield five-membered chelate complexes [(NwedgeN)Pd(kappa2-CH(Et)(OC(O)-CH3))]+, 4, and [(NwedgeN)Pd(kappa2 CH(Et)(OC(O)CF3))]+, 6. When VA is added to [(NwedgeN)Ni(CH3)]+, an equilibrium mixture of an eta2 olefin complex, 8c, and a kappa-oxygen complex, 8o, results. Insertion occurs from the eta2 olefin complex, 8c (DeltaG++ = 15.5 kcal/mol, -51 degrees C), in both a 2,1 and a 1,2 fashion to generate a mixture of five- and six-membered chelates, 9(2,1) and 9(1,2). VA(f) inserts into the Ni-CH3 bond (-80 degrees C) to form a five-membered chelate with no detectable intermediate. Thermolysis of the Pd chelates results in beta-acetate elimination from 4 (DeltaG++ = 25.5 kcal/mol, 60 degrees C) and beta-trifluoroacetate elimination from 6 (DeltaG = 20.5 kcal/mol, 10 degrees C). The five-membered Ni chelate, 9(2,1), is quite stable at room temperature, but the six-membered chelate, 9(1,2), undergoes beta-elimination at -34 degrees C. Treatment of the OAc(f) containing Pd chelate 6 with ethylene results in complete opening to the pi complex [(NwedgeN)Pd(kappa2-CH(Et)(OAc(f)))(CH2CH2)]+ (OAc(f) = OC(O)CF3), 18, while reaction of the OAc containing Pd chelate 4 with ethylene establishes an equilibrium between 4 and the open form 16, strongly favoring the closed chelate 4 (DeltaH = -4.1 kcal/mol, DeltaS = -23 eu, K = 0.009 M(-1) at 25 degrees C). The open chelates undergo migratory insertion at much slower rates relative to those of the simple (NwedgeN)Pd(CH3)(CH2CH2)+ analogue. These quantitative studies provide an explanation for the behavior of VA and VA(f) in attempted copolymerizations with ethylene. PMID- 15810849 TI - Rigid MIIL2Gd2III (M = Fe, Ru) complexes of a terpyridine-based heteroditopic chelate: a class of candidates for MRI contrast agents. AB - Rigid chelates of high-molecular weight, [M(tpy-DTTA)2]6- (M = Fe, Ru), are obtained upon self-assembly around one M(II) ion of two terpyridine-based molecules substituted in the 4'-position with the polyaminocarboxylate diethylenetriamine-N,N,N'',N''-tetraacetate, tpy-DTTA4-. The protonation constants of tpy-DTTA4- (log K1 = 8.65(4), log K2 = 7.63(4), log K3 = 5.25(6), log K4 = 3.30(7)) and [Fe(tpy-DTTA)2]6- (log K1 = 8.40(4), log K2 = 7.26(4)) have been determined by potentiometry, 1H NMR and UV-vis titrations. The thermodynamic stability constant log K(GdL) of [Fe(tpy-DTTA)2Gd2(H2O)4] measured at 25 degrees C by potentiometry is 10.87. This relatively low value is due to the direct linkage of the polyaminocarboxylate part to the electron-withdrawing terpyridine. UV-vis absorbance spectra of [M(tpy-DTTA)2Gd2(H2O)4] and 1H NMR spectra of [M(tpy DTTA)2Eu2(H2O)4] revealed similar solution behavior of the Fe and Ru complexes. An I(d) water-exchange mechanism (DeltaV++ = +6.8 +/- 1 cm3 mol(-1)) with a rate constant of k(ex)298 = (5.1 +/- 0.3) x 10(6) s(-1) has been found for [Fe(tpy DTTA)2Gd2(H2O)4] by 17O NMR. A slow rotational correlation time (tau(RO) = 410 +/ 10 ps) and the presence of two water molecules (q = 2) in the coordination inner sphere of each Gd(III) ion have also been determined for this complex. A remarkably high relaxivity has been observed for both [M(tpy-DTTA)2Gd2(H2O)4] complexes (at 20 MHz and 37 degrees C, r(1) = 15.7 mM(-1) s(-1) for the Fe complex, and r(1) = 15.6 mM(-1) s(-1) for the Ru complex). PMID- 15810850 TI - Dye sensitization of the anatase (101) crystal surface by a series of dicarboxylated thiacyanine dyes. AB - Dye sensitization of the single crystal anatase (101) surface was studied using a structurally similar series of dicarboxylated thiacyanine dyes that bind to the oxide surface through their carboxylate groups. An ultraviolet (UV) light treatment of the anatase (101) surfaces, immediately prior to dye adsorption, improved both the reproducibility of dye coverage and the incident photon-to current efficiencies (IPCE) for sensitization. The UV treatment does not pit or roughen the anatase surface and results in high IPCEs of more than 1% in some cases and absorbed photon current efficiencies (APCE) from 5 to 100%. The photocurrent spectra showed features associated with surface-bound dye monomers and H-dimers that could be followed as a function of the dye surface coverage. Models for the surface structures of the adsorbed dye layers that are consistent with the measurements are presented, along with a discussion of adsorption isotherms. PMID- 15810851 TI - Luminescent ruthenium(II)- and rhenium(I)-diimine wires bind nitric oxide synthase. AB - Ru(II)- and Re(I)-diimine wires bind to the oxygenase domain of inducible nitric oxide synthase (iNOSoxy). In the ruthenium wires, [Ru(L)2L']2+, L' is a perfluorinated biphenyl bridge connecting 4,4'-dimethylbipyridine to a bulky hydrophobic group (adamantane, 1), a heme ligand (imidazole, 2), or F (3). 2 binds in the active site of the murine iNOSoxy truncation mutants Delta65 and Delta114, as demonstrated by a shift in the heme Soret from 422 to 426 nm. 1 and 3 also bind Delta65 and Delta114, as evidenced by biphasic luminescence decay kinetics. However, the heme absorption spectrum is not altered in the presence of 1 or 3, and Ru-wire binding is not affected by the presence of tetrahydrobiopterin or arginine. These data suggest that 1 and 3 may instead bind to the distal side of the enzyme at the hydrophobic surface patch thought to interact with the NOS reductase module. Complexes with properties similar to those of the Ru-diimine wires may provide an effective means of NOS inhibition by preventing electron transfer from the reductase module to the oxygenase domain. Rhenium-diimine wires, [Re(CO)3L1L1']+, where L1 is 4,7-dimethylphenanthroline and L1' is a perfluorinated biphenyl bridge connecting a rhenium-ligated imidazole to a distal imidazole (F8bp-im) (4) or F (F9bp) (5), also form complexes with Delta114. Binding of 4 shifts the Delta114 heme Soret to 426 nm, demonstrating that the terminal imidazole ligates the heme iron. Steady-state luminescence measurements establish that the 4:Delta114 dissociation constant is 100 +/- 80 nM. Re-wire 5 binds Delta114 with a K(d) of 5 +/- 2 microM, causing partial displacement of water from the heme iron. Our finding that both 4 and 5 bind in the NOS active site suggests novel designs for NOS inhibitors. Importantly, we have demonstrated the power of time-resolved FET measurements in the characterization of small molecule:protein interactions that otherwise would be difficult to observe. PMID- 15810852 TI - Theoretical study on the inhibition of ribonucleotide reductase by 2'-mercapto-2' deoxyribonucleoside-5'-diphosphates. AB - Ribonucleotide reductase (RNR) is responsible for the reduction of ribonucleotides into the correspondent 2'-deoxyribonucleotides in the only physiological process that yields the monomers of DNA. The enzyme has thus become an attractive target for chemotherapies that fight proliferation-based diseases, specifically cancer and infections by some viruses and parasites. 2'-Mercapto-2' deoxyribonucleoside-5'-diphosphates (SHdNDP) are mechanism-based inhibitors of RNR and therefore potential chemotherapeutic agents for those indications. Previous experimental studies established the in vitro and in vivo activity of SHdNDP. In the in vitro studies, it was observed that the activity was dependent on the oxidative status of the medium, with the inactivation of RNR only occurring when molecular oxygen was available. To better understand the mechanism involved in RNR inactivation by SHdNDP, we performed theoretical calculations on the possible reactions between the inhibitors and the RNR active site. As a result, we propose the possible mechanistic pathways for the chemical events that occur in the absence and in the presence of O2. They correspond to a refinement and a complement of those proposed in the literature. PMID- 15810853 TI - Anisotropic local motions and location of amide protons in proteins. AB - A new method has been developed to obtain dynamic and structural information about peptide planes in proteins by a combination of measurements of weak short range cross-correlation rates R(H(N)N/NC') that are due to concerted fluctuations of the H(N)-N and N-C' dipole-dipole interactions and stronger long-range cross correlation rates R(C'H(N)/H(N)N) and R(NH(N)/H(N)C(alpha)). The rates were interpreted using the axially symmetric Gaussian axial fluctuation model (GAF). The oscillation amplitudes as well as the positions of H(N) atoms with respect to peptide planes in ubiquitin were determined. Most N-H(N) bonds were found not to lie exactly along the bisector of the N-C' and N-C(alpha) bonds but to be slightly tilted toward the carbon-terminal side of the peptide. PMID- 15810854 TI - Potentiometric, electronic structural, and ground- and excited-state optical properties of conjugated bis[(porphinato)zinc(II)] compounds featuring proquinoidal spacer units. AB - We report the synthesis, optical, electrochemical, electronic structural, and transient optical properties of conjugated (porphinato)zinc(II)-spacer (porphinato)zinc(II) (PZn-Sp-PZn) complexes that possess intervening conjugated Sp structures having varying degrees of proquinoidal character. These supermolecular PZn-Sp-PZn compounds feature Sp moieties {(4,7 diethynylbenzo[c][1,2,5]thiadiazole (E-BTD-E), 6,13-diethynylpentacene (E-PC-E), 4,9-diethynyl-6,7-dimethyl[1,2,5]thiadiazolo[3,4-g]quinoxaline (E-TDQ-E), and 4,8 diethynylbenzo[1,2-c:4,5-c']bis([1,2,5]thiadiazole) (E-BBTD-E)} that regulate frontier orbital energy levels and progressively increase the extent of the quinoidal resonance contribution to the ground and electronically excited states, augmenting the magnitude of electronic communication between terminal (5,-10,20 di(aryl)porphinato)zinc(II) units, relative to that evinced for a bis[(5,5', 10,20-di(aryl)porphinato)zinc(II)]butadiyne benchmark (PZnE-EPZn). Electronic absorption spectra show significant red-shifts of the respective PZn-Sp-PZn x polarized Q state (S0 --> S1) transition manifold maxima (240-4810 cm(-1)) relative to that observed for PZnE-EPZn. Likewise, the potentiometrically determined PZn-Sp-PZn HOMO-LUMO gaps (E1/2(0/+) - E1/2(-/0)) display correspondingly diminished energy separations that range from 1.88 to 1.11 eV relative to that determined for PZnE-EPZn (2.01 eV). Electronic structure calculations provide insight into the origin of the observed PZn-Sp-PZn electronic and optical properties. Pump-probe transient spectral data for these PZn-Sp-PZn supermolecules demonstrate that the S1 --> S(n) transition manifolds of these species span an unusually broad spectral domain of the NIR. Notably, the absorption maxima of these S1 --> S(n) manifolds can be tuned over a 1000-1600 nm spectral region, giving rise to intense excited-state transitions approximately 4000 cm(-1) lower in energy than that observed for the analogous excited-state absorption maximum of the PZnE-EPZn benchmark; these data highlight the unusually large quinoidal resonance contribution to the low-lying electronically excited singlet states of these PZn-Sp-PZn species. The fact that the length scales of the PZn-Sp-PZn species (approximately 25 angstrom) are small with respect to those of classic conducting polymers, yet possess NIR S1 --> S(n) manifold absorptions lower in energy, underscore the unusual electrooptic properties of these conjugated structures. PMID- 15810855 TI - A diameter-selective attack of metallic carbon nanotubes by nitronium ions. AB - We have found a method for a diameter-selective removal of metallic single-walled carbon nanotubes (m-SWCNTs) from semiconducting (s-) ones by stirring or sonicating SWCNT powder in tetramethylene sulfone (TMS)/chloroform solution with nitronium hexafluoroantimonate (NO2SbF6: NHFA) and nitronium tetrafluoroborate (NO2BF4: NTFB). Positively charged nitronium ions (NO2+) were intercalated into nanotube bundles, where the intercalation was promoted also by the counterions. Nitronium ions selectively attacked the sidewall of the m-SWCNTs due to the abundant presence of electron density at the Fermi level, thus yielding stronger binding energy as compared to the counterpart s-SWCNTs. The s-SWCNTs were left on the filter after filtration, whereas the m-SWCNTs were disintegrated and drained away as amorphous carbons. The effectiveness of removing m-SWCNTs was confirmed by the resonant Raman spectra and absorption spectra. PMID- 15810856 TI - Reactions of d0 group 4 amides with dioxygen. Preparation of unusual oxo aminoxy complexes and theoretical studies of their formation. AB - Reactions of d0 amides M(NMe2)4 (M = Zr, 1; Hf, 2) with O2 have been found to yield unusual trinuclear oxo aminoxide complexes M3(NMe2)6(mu-NMe2)3(mu3-O)(mu3 ONMe2) (M = Zr, 3; Hf, 4) in high yields. Tetramethylhydrazine Me2N-NMe2 was also observed in the reaction mixtures. Crystal structures of 3 and 4 have been determined. Density functional theory calculations have been performed to explore the mechanistic pathways in the reactions of model complexes Zr(NR2)4 (R = H, 5; Me, 1) and [Zr(NR2)4]2 (R = H, 5a; Me, 1a) with triplet O2. Monomeric and dimeric reaction pathways in the formation of the Zr complex 3 are proposed. PMID- 15810857 TI - Synthesis and reactivity of a (mu-1,1-hydroperoxo)(mu-hydroxo)dicopper(II) complex: ligand hydroxylation by a bridging hydroperoxo ligand. AB - A new tetradentate tripodal ligand (L3) containing sterically bulky imidazolyl groups was synthesized, where L3 is tris(1-methyl-2-phenyl-4 imidazolylmethyl)amine. Reaction of a bis(mu-hydroxo)dicopper(II) complex, [Cu2(L3)2(OH)2]2+ (1), with H2O2 in acetonitrile at -40 degrees C generated a (mu 1,1-hydroperoxo)dicopper(II) complex [Cu2(L3)2(OOH)(OH)]2+ (2), which was characterized by various physicochemical measurements including X-ray crystallography. The crystal structure of 2 revealed that the complex cation has a Cu2(mu-1,1-OOH)(mu-OH) core and each copper has a square pyramidal structure having an N3O2 donor set with a weak ligation of a tertiary amine nitrogen in the apex. Consequently, one pendant arm of L3 in 2 is free from coordination, which produces a hydrophobic cavity around the Cu2(mu-1,1-OOH)(mu-OH) core. The hydrophobic cavity is preserved by hydrogen bondings between the hydroperoxide and the imidazole nitrogen of an uncoordinated pendant arm in one side and the hydroxide and the imidazole nitrogen of an uncoordinated pendant arm in the other side. The hydrophobic cavity significantly suppresses the H/D and 16O/18O exchange reactions in 2 compared to that in 1 and stabilizes the Cu2(mu-1,1 OOH)(mu-OH) core against decomposition. Decomposition of 2 in acetonitrile at 0 degrees C proceeded mainly via disproportionation of the hydroperoxo ligand and reduction of 2 to [Cu(L3)]+ by hydroperoxo ligand. In contrast, decomposition of a solid sample of 2 at 60 degrees C gave a complex having a hydroxylated ligand [Cu2(L3)(L3-OH)(OH)2]2+ (2-(L3-OH)) as a main product, where L3-OH is an oxidized ligand in which one of the methylene groups of the pendant arms is hydroxylated. ESI-TOF/MS measurement showed that complex 2-(L3-OH) is stable in acetonitrile at -40 degrees C, whereas warming 2-(L3-OH) at room temperature resulted in the N dealkylation from L3-OH to give an N-dealkylated ligand, bis(1-methyl-2-phenyl-4 imidazolylmethyl)amine (L2) in approximately 80% yield based on 2, and 1-methyl-2 phenyl-4-formylimidazole (Phim-CHO). Isotope labeling experiments confirmed that the oxygen atom in both L3-OH and Phim-CHO come from OOH. This aliphatic hydroxylation performed by 2 is in marked contrast to the arene hydroxylation reported for some (mu-1,1-hydroperoxo)dicopper(II) complexes with a xylyl linker. PMID- 15810858 TI - The final catalytic step of cytochrome p450 aromatase: a density functional theory study. AB - B3LYP density functional theory calculations are used to unravel the mysterious third step of aromatase catalysis. The feasibility of mechanisms in which the reduced ferrous dioxygen intermediate mediates androgen aromatization is explored and determined to be unlikely. However, proton-assisted homolysis of the peroxo hemiacetal intermediate to produce P450 compound I and the C19 gem-diol likely proceeds with a low energetic barrier. Mechanisms for the aromatization and deformylation sequence which are initiated by 1beta-hydrogen atom abstraction by P450 compound I are considered. 1beta-Hydrogen atom abstraction from substrates in the presence of the 2,3-enol encounters strikingly low barriers (5.3-7.8 kcal/mol), whereas barriers for this same process rise to 17.0-27.1 kcal/mol in the keto tautomer. Transition states for 1beta-hydrogen atom abstraction from enolized substrates in the presence of the 19-gem-diol decayed directly to the experimentally observed products. If the C19 aldehyde remains unhydrated, aromatization occurs with concomitant decarbonylation and therefore does not support dehydration of the C19 aldehyde prior to the final catalytic step. On the doublet surface, the transition state connects to a potentially labile 1(10) dehydrogenated product, which may undergo rapid aromatization, as well as formic acid. Ab initio molecular dynamics confirmed that the 1beta-hydrogen atom abstraction and deformylation or decarbonylation occur in a nonsynchronous, coordinated manner. These calculations support a dehydrogenase behavior of aromatase in the final catalytic step, which can be summarized by 1beta-hydrogen atom abstraction followed by gem-diol deprotonation. PMID- 15810859 TI - Anhydrous and water-assisted proton mobility in phosphotungstic acid. AB - Nonlocal gradient-corrected density functional theoretical calculations were used to determine the energetics associated with proton migration in phosphotungstic acid. The activation energy for anhydrous proton hopping between two oxygen atoms on the exterior of the molecular Keggin unit was calculated to be 103.3 kJ mol( 1). The quantum-tunneling effect on the rate of proton movement was determined using semiclassical transition-state theory and was found to be a major contributor to the overall rate of proton movement at temperatures below approximately 350 K. The adsorption of water on an acidic proton decreases the activation barrier for hopping to 11.2 kJ mol(-1) by facilitating proton transfer along hydrogen bonds. The overall rate constant for proton hopping was determined as a function of temperature and water partial pressure. Small amounts of water greatly enhance the overall rate of proton movement. PMID- 15810860 TI - Temperature dependence of quantum effects in liquid water. AB - Quantum and classical simulations are carried out on model water systems over a wide range of temperatures, from 100 to -35 degrees C. A detailed examination of the equilibrium and dynamical properties of liquid water is presented, together with a discussion of the interplay between quantum mechanical tunneling and dynamics. The study shows that quantum effects are essential for a description of the dynamical behavior of liquid water, particularly in the low-temperature (supercooled) region. The similarities and differences between quantum effects and the effects associated with increasing the temperature are explicitly characterized. PMID- 15810861 TI - Oxygen electroreduction through a superoxide intermediate on bi-modified Au surfaces. AB - The mechanism of the electroreduction of oxygen on bare and Bi-submonolayer modified Au(111) surfaces is examined using surface enhanced Raman scattering (SERS) measurements along with detailed density functional theory (DFT) calculations. The spectroscopy reveals the presence of superoxide-level species at potentials where oxygen is reduced. These species are not present in solutions absent either oxygen or Bi at these potentials. The spectroscopy also reveals the presence of Bi-OH species which are associated with peroxide reduction. Detailed calculations show oxygen associates much more strongly with Bi in the (2 x 2) configuration on Au(111) relative to the bare Au surface. Additionally, the O-O bond is elongated following O2 association, which follows as a consequence of Bi O bond formation and partial oxidation of the Bi adatom. These results show for the first time that the four-electron electroreduction of oxygen electroreduction occurs via a series pathway on the Bi-modified surface in acid solution. PMID- 15810862 TI - Glutathione-protected gold clusters revisited: bridging the gap between gold(I) thiolate complexes and thiolate-protected gold nanocrystals. AB - Small gold clusters (approximately 1 nm) protected by molecules of a tripeptide, glutathione (GSH), were prepared by reductive decomposition of Au(I)-SG polymers at a low temperature and separated into a number of fractions by polyacrylamide gel electrophoresis (PAGE). Chemical compositions of the fractionated clusters determined previously by electrospray ionization (ESI) mass spectrometry (Negishi, Y. et al. J.Am. Chem. Soc. 2004, 126, 6518) were reassessed by taking advantage of freshly prepared samples, higher mass resolution, and more accurate mass calibration; the nine smallest components are reassigned to Au10(SG)10, Au15(SG)13, Au18(SG)14, Au22(SG)16, Au22(SG)17, Au25(SG)18, Au29(SG)20, Au33(SG)22, and Au39(SG)24. These assignments were further confirmed by measuring the mass spectra of the isolated Au:S(h-G) clusters, where h-GSH is a homoglutathione. It is proposed that a series of the isolated Au:SG clusters corresponds to kinetically trapped intermediates of the growing Au cores. The relative abundance of the isolated clusters was correlated well with the thermodynamic stabilities against unimolecular decomposition. The electronic structures of the isolated Au:SG clusters were probed by X-ray photoelectron spectroscopy (XPS) and optical spectroscopy. The Au(4f) XPS spectra illustrate substantial electron donation from the gold cores to the GS ligands in the Au:SG clusters. The optical absorption and photoluminescence spectra indicate that the electronic structures of the Au:SG clusters are well quantized; embryos of the sp band of the bulk gold evolve remarkably depending on the number of the gold atoms and GS ligands. The comparison of these spectral data with those of sodium Au(I) thiomalate and 1.8 nm Au:SG nanocrystals (NCs) reveals that the subnanometer sized Au clusters thiolated constitute a distinct class of binary system which lies between the Au(I)-thiolate complexes and thiolate-protected Au NCs. PMID- 15810864 TI - Effects of omega-3 fatty acids on organ transplantation. PMID- 15810865 TI - Trends in the management of type 2 diabetes: an emerging role for insulin. AB - OBJECTIVE: This review is intended to explore the pathophysiology of type 2 diabetes, examine the role of insulin as a means of achieving glycemic control in people with type 2 diabetes, and provide a practical approach for insulin use in type 2 diabetes in the managed care setting. DATA SOURCES: This manuscript is based on the results of a MEDLINE literature search and presentations by the authors at a symposium titled, "Emerging Changes in Diabetes Management," that took place on October, 14, 2004, at the Academy of Managed Care Pharmacy's 2004 Educational Conference in Baltimore, Maryland. CONCLUSIONS: Despite advances in oral treatment, type 2 diabetes remains a substantial source of microvascular and macrovascular complications that cause unacceptable levels of morbidity, mortality, and cost. Accumulating clinical evidence suggests that insulin treatment, both basal and prandial, can advance the treatment of type 2 diabetes and reduce the risks for serious sequelae by providing consistent and optimal glycemic control. By more closely mimicking the actions of endogenous insulin, in terms of onset and duration of action, insulin analogues offer clear advantages over their regular insulin counterparts. PMID- 15810866 TI - Empirical and conceptual problems with longitudinal trait-state models: introducing a trait-state-occasion model. AB - The latent trait-state-error model (TSE) and the latent state-trait model with autoregression (LST-AR) represent creative structural equation methods for examining the longitudinal structure of psychological constructs. Application of these models has been somewhat limited by empirical or conceptual problems. In the present study, Monte Carlo analysis revealed that TSE models tend to generate improper solutions when N is too small, when waves are too few, and when occasion factor stability is either too large or too small. Mathematical analysis of the LST-AR model revealed its limitation to constructs that become more highly auto correlated over time. The trait-state-occasion model has fewer empirical problems than does the TSE model and is more broadly applicable than is the LST-AR model. PMID- 15810867 TI - Investigating population heterogeneity with factor mixture models. AB - Sources of population heterogeneity may or may not be observed. If the sources of heterogeneity are observed (e.g., gender), the sample can be split into groups and the data analyzed with methods for multiple groups. If the sources of population heterogeneity are unobserved, the data can be analyzed with latent class models. Factor mixture models are a combination of latent class and common factor models and can be used to explore unobserved population heterogeneity. Observed sources of heterogeneity can be included as covariates. The different ways to incorporate covariates correspond to different conceptual interpretations. These are discussed in detail. Characteristics of factor mixture modeling are described in comparison to other methods designed for data stemming from heterogeneous populations. A step-by-step analysis of a subset of data from the Longitudinal Survey of American Youth illustrates how factor mixture models can be applied in an exploratory fashion to data collected at a single time point. PMID- 15810868 TI - Meta-analytic structural equation modeling: a two-stage approach. AB - To synthesize studies that use structural equation modeling (SEM), researchers usually use Pearson correlations (univariate r), Fisher z scores (univariate z), or generalized least squares (GLS) to combine the correlation matrices. The pooled correlation matrix is then analyzed by the use of SEM. Questionable inferences may occur for these ad hoc procedures. A 2-stage structural equation modeling (TSSEM) method is proposed to incorporate meta-analytic techniques and SEM into a unified framework. Simulation results reveal that the univariate-r, univariate-z, and TSSEM methods perform well in testing the homogeneity of correlation matrices and estimating the pooled correlation matrix. When fitting SEM, only TSSEM works well. The GLS method performed poorly in small to medium samples. PMID- 15810869 TI - A latent markov model for the analysis of longitudinal data collected in continuous time: states, durations, and transitions. AB - Markov models provide a general framework for analyzing and interpreting time dependencies in psychological applications. Recent work extended Markov models to the case of latent states because frequently psychological states are not directly observable and subject to measurement error. This article presents a further generalization of latent Markov models to allow for the analysis of rating data that are collected at arbitrary points in time. This extension offers new ways of investigating change processes by focusing explicitly on the durations that are spent in latent states. In an experience sampling application the author shows that such duration analyses can provide valuable insights about chronometric features of emotions. PMID- 15810870 TI - Using data augmentation to obtain standard errors and conduct hypothesis tests in latent class and latent transition analysis. AB - Latent class analysis (LCA) provides a means of identifying a mixture of subgroups in a population measured by multiple categorical indicators. Latent transition analysis (LTA) is a type of LCA that facilitates addressing research questions concerning stage-sequential change over time in longitudinal data. Both approaches have been used with increasing frequency in the social sciences. The objective of this article is to illustrate data augmentation (DA), a Markov chain Monte Carlo procedure that can be used to obtain parameter estimates and standard errors for LCA and LTA models. By use of DA it is possible to construct hypothesis tests concerning not only standard model parameters but also combinations of parameters, affording tremendous flexibility. DA is demonstrated with an example involving tests of ethnic differences, gender differences, and an Ethnicity x Gender interaction in the development of adolescent problem behavior. PMID- 15810871 TI - Global, local, and graphical person-fit analysis using person-response functions. AB - Person-fit statistics test whether the likelihood of a respondent's complete vector of item scores on a test is low given the hypothesized item response theory model. This binary information may be insufficient for diagnosing the cause of a misfitting item-score vector. The authors propose a comprehensive methodology for person-fit analysis in the context of nonparametric item response theory. The methodology (a) includes H. Van der Flier's (1982) global person-fit statistic U3 to make the binary decision about fit or misfit of a person's item score vector, (b) uses kernel smoothing (J. O. Ramsay, 1991) to estimate the person-response function for the misfitting item-score vectors, and (c) evaluates unexpected trends in the person-response function using a new local person-fit statistic (W. H. M. Emons, 2003). An empirical data example shows how to use the methodology for practical person-fit analysis. PMID- 15810872 TI - The role of item extremity in the emergence of keying-related factors: an exploration with the life orientation test. AB - Keying-related factors in psychological scales are variously interpreted substantively or as products of violations of the assumptions underlying item keying. The present study investigated whether the extremity of the wording of items may contribute to the emergence of item-keying factors in a commonly used psychological scale. Respondents (N = 277) completed the Life Orientation Test (M. F. Scheier & C. S. Carver, 1985) in either its original or modified, more moderately worded form. Results indicate that the interaction of item extremity and item keying significantly affected subscale means and, more important, that the more moderately worded scale was substantially more unidimensional. Results are explained partially through the association of lesser and greater extremity with the tendency for some respondents to agree or disagree with items irrespective of keying direction. These results, although demonstrated in only 1 scale, have potential relevance to any scale comprising positive and negative items. PMID- 15810874 TI - Exploiting novel cell cycle targets in the development of anticancer agents. AB - In this review we provide a brief background on the cell cycle and then focus on two novel and emerging areas of cell cycle research that may prove to have significant relevance to the development of novel anticancer agents. In particular, we review the emerging evidence to suggest that histone deacetylase inhibitors may possess cancer cell-specific cytotoxicity due to their ability to target a novel G2/M checkpoint. We also review the recent literature supporting the proposition that inhibition of E2F activity in epithelial cancer cells may prove to be a useful differentiation therapy that operates via cell cycle dependent and cell cycle-independent mechanisms. PMID- 15810875 TI - NO news is not necessarily good news in cancer. AB - The diatomic radical nitric oxide has been the focus of numerous studies involved with every facet of cancer. It has been implicated in carcinogenesis, progression, invasion, metastasis, angiogenesis, escape from immune surveillance, and modulation of therapeutic response. In recent years, an increasing number of studies have suggested the possible involvement of nitric oxide in multiple cancer types, including melanoma. It is perhaps not surprising that conflicting viewpoints have arisen as to whether nitric oxide is beneficial or deleterious in cancer. However, it has become clear that nitric oxide possesses modulatory properties in a number of signal transduction pathways that depend on concentration and context. Our laboratory has shown that tumor expression of inducible nitric oxide synthase in melanoma patients results in poor survival. Furthermore, we demonstrated that the removal of endogenous nitric oxide in melanoma cell lines led to increased sensitivity to cisplatin-induced apoptosis in a p53-dependent manner. Others have shown anti-apoptotic properties of NO in melanoma cells. However, several studies also suggest that NO can inhibit metastasis and diminish resistance. Despite the apparently conflicting observations, it is evident that NO is involved in melanoma pathology. The purpose of this review is to summarize the current literature relating to the role of NO in cancer with particular emphasis on its relevance to therapeutic resistance in melanoma. Recent evidence suggests the involvement of an intricate and complex interplay between reactive nitrogen species and reactive oxygen species. The importance of nitric oxide and its balance with other oxidative agents in the regulation of cancer cell response to therapies will be discussed. This balance may serve as an important focal point in determining patient response to therapy. The ability to control this balance could significantly influence outcome. PMID- 15810876 TI - Induction of apoptosis by curcumin and its implications for cancer therapy. AB - Curcumin (diferuloyl methane), the yellow pigment in turmeric (Curcuma longa), is a potent chemopreventive agent that inhibits proliferation of cancer cells by arresting them at various phases of the cell cycle depending upon the cell type. Curcumin-induced apoptosis mainly involves the mitochondria-mediated pathway in various cancer cells of different tissues of origin. In some cell types like thymocytes, curcumin induces apoptosis-like changes whereas in many other normal and primary cells curcumin is either inactive or inhibits proliferation, but does not appear to induce apoptosis. These together with reports that curcumin protects cells against apoptosis induced by other agents, underscore the need for further understanding of the multiple mechanisms of cell death unleashed by curcumin. Tumor cells often evade apoptosis by expressing several antiapoptotic proteins, down-regulation and mutation of proapoptotic genes and alterations in signaling pathways that give them survival advantage and thereby allow them to resist therapy-induced apoptosis. Many researchers including ourselves, have demonstrated the involvement of several pro and antiapoptotic molecules in curcumin-induced apoptosis, and ways to sensitize chemoresistant cancer cells to curcumin treatment. This review describes the mechanisms of curcumin-induced apoptosis currently known, and suggests several potential strategies that include down-regulation of antiapoptotic proteins by antisense oligonucleotides, use of proapoptotic peptides and combination therapy, and other novel approaches against chemoresistant tumors. Several factors including pharmacological safety, scope for improvement of structure and function of curcumin and its ability to attack multiple targets are in favor of curcumin being developed as a drug for prevention and therapy of various cancers. PMID- 15810877 TI - Treatment modalities for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, accounting for approximately 5 to 10% of all cancers. It is estimated to cause approximately 1 million deaths annually. Currently, no adjuvant or palliative treatment modalities have been conclusively shown to prolong survival in HCC. Despite the high mortality and frequency of this cancer, surgical resection is an option for only a small proportion of patients, less than 18%. Liver cirrhosis is the most common cause of HCC and necessitates the preservation as much liver as possible, resulting in local ablation, intra-arterial and systemic treatments being major therapeutic modalities. Through better understanding of the molecular basis of hepatocarcinogenesis, new preventative and treatment modalities have recently emerged. This article reviews the current treatment options and new therapeutic advances for HCC including antiangiogenesis therapy, targeted therapy and antisense gene targeting. Future clinical trials and research will help to evaluate and improve both systemic and targeted molecular therapies for this complex disease. PMID- 15810878 TI - The radionuclide molecular imaging and therapy of neuroendocrine tumors. AB - Neuroendocrine tumors (NETs) represent a large group of neoplasms deriving from pluripotent stem cells or from differentiated neuroendocrine cells that are characterized by the expression of different peptides and biogenic amines. These rare tumors tend to grow slowly and are notoriously difficult to localize, at least in the early stages. Diagnostics involve blood, urine and biochemical examination as well as imaging modalities. Imaging is achieved by a variety of techniques such as radiological morphological imaging methods, for example, sonography, computerized tomography (CT)/magnetic resonance imaging (MRI), angiography and finally, nuclear functional imaging methods such as metaiodobenzylguanidine (MIBG), somatostatin receptor scintigraphy (SRS), vasoactive intestinal peptide receptor scintigraphy (VIPRS) and positron emission tomography (PET) using (18)F labeled deoxyglucose (FDG) and fluorinated dihydroxyphenylalanine ((18)F-DOPA) as a radioisotopic marker. (131)I-labeled MIBG is a well-established radiopharmaceutical for localization and therapy of phechromocytoma and paraganglioma. The majority of neuroendocrine tumors possess a high density of somatostatin receptors. This observation provided the basis for the development of various radiolabeled somatostatin peptide analogs as imaging agents and therapeutics in nuclear medicine. FDG-PET is now performed in a wide variety of tumors and indications, including diagnosis, staging, re-staging and evaluation of the response to treatment. (18)F-DOPA-PET may be useful if (18)F FDG-PET scan result is negative. (99m)Tc-pentavalent dimercaptosuccinic acid ((99m)Tc-DMSA-V) or (99m)Tc sestamibi ((99m)Tc-MIBI) or (99m)Tc-tetrofosmin is used only for diagnosis of certain NETs such as medullary thyroid cancer. The expiences with other nuclear medicinie imaging and therapy modalities such as cholecystokinin (CCK)-B/gastrin-receptors, bombesin/gastrin-releasing peptide receptor scintigraphy are still limited, and further clinical studies are needed. The studies using vascular endothelial growth factor (VEGF) for tumor angiogenesis imaging, annexin-V for imaging apoptosis and agents for hypoxia imaging are still in an early stage and the clinical role for these agents needs to be defined. In conclusion, no single imaging technique identifies all the metastatic sites of NETs. The best results may be obtained with a combination of functional imaging such as PET or/and SRS and morphologic imaging with CT and/or MR imaging. Many molecular imaging and therapy modalities fur NETs are recently under investigation or being developed, the usefulness of these modalities, however, has to be evaluated by well-designed and multicentre studies. PMID- 15810879 TI - Englitazone administration to late pregnant rats produces delayed body growth and insulin resistance in their fetuses and neonates. AB - The level of maternal circulating triacylglycerols during late pregnancy has been correlated with the mass of newborns. PPARgamma (peroxisome-proliferator activated receptor gamma) ligands, such as TZDs (thiazolidinediones), have been shown to reduce triacylglycerolaemia and have also been implicated in the inhibition of tissue growth and the promotion of cell differentiation. Therefore TZDs might control cell proliferation during late fetal development and, by extension, body mass of pups. To investigate the response to EZ (englitazone), a TZD, on perinatal development, 0 or 50 mg of englitazone/kg of body mass was given as an oral dose to pregnant rats daily from day 16 of gestation until either day 20 for the study of their fetuses, or until day 21 of gestation for the study of neonates. EZ decreased maternal triacylglycerol levels at day 20 of gestation and neonatal mass, but not fetal mass. Fetuses and neonates from EZ treated mothers exhibited high levels of insulin and were found to be hyperglycaemic. The apparent insulin-resistant state in neonates from EZ-treated pregnant rats was corroborated, since they showed higher plasma NEFA [non esterified ('free') fatty acid] levels, ketonaemia and liver LPL (lipoprotein lipase) activity and lower plasma IGF-I (type 1 insulin-like growth factor) levels, in comparison with those from control mothers. Moreover, at the molecular level, an increase in Akt phosphorylation was found in the liver of neonates from EZ-treated mothers, which confirms that the insulin pathway was negatively affected. Thus the response of fetuses and neonates to maternal antidiabetic drug treatment is the opposite of what would be expected, and can be justified by the scarce amount of adipose tissue impeding a normal response to PPARgamma ligands and by hyperinsulinaemia as being responsible for a major insulin-resistant condition. PMID- 15810880 TI - Cloning of ABCA17, a novel rodent sperm-specific ABC (ATP-binding cassette) transporter that regulates intracellular lipid metabolism. AB - The A subclass of the ABC (ATP-binding cassette) transporter superfamily has a structural feature that distinguishes it from other ABC transporters, and is proposed to be involved in the transmembrane transport of endogenous lipids. Here we have cloned mouse and rat full-length cDNAs of ABCA17, a novel ABC transporter belonging to the A subclass. Mouse and rat ABCA17 proteins comprise 1733 and 1773 amino acid residues respectively, having 87.3% amino acid identity; mouse ABCA17 has amino acid identities of 55.3% and 36.7% with mouse ABCA3 and sea urchin ABCA respectively. RNA blot and quantitative real-time PCR analyses showed that ABCA17 mRNA is expressed exclusively in the testis. Examination of testis by in situ hybridization showed that ABCA17 mRNA is expressed in germ cells, mainly spermatocytes, in the seminiferous tubule. Immunoblot analysis using a specific antibody showed that ABCA17 is a protein of 200 kDa, and immunohistochemical analysis demonstrated that the protein is detected in the anterior head of sperm and elongated spermatids. ABCA17 was localized in the endoplasmic reticulum in transiently transfected HEK293 cells. Metabolic labelling analysis showed that intracellular esterified lipids, including cholesteryl esters, fatty acid esters and triacylglycerols, were significantly decreased in HEK293 cells stably expressing ABCA17 compared with untransfected cells. These results suggest that ABCA17 may play a role in regulating lipid composition in sperm. PMID- 15810881 TI - Progress in epidermolysis bullosa: the phenotypic spectrum of plectin mutations. AB - Plectin, a large multidomain adhesive protein with versatile binding functions, is expressed in a number of tissues and cell types. In the skin, plectin is a critical component of hemidesmosomes, interacting with keratin intermediate filaments and beta4 integrin. Mutations in the plectin gene (PLEC1) result in fragility of skin, demonstrating blister formation at the level of hemidesmosomes. These blistering disorders belong to the spectrum of epidermolysis bullosa (EB) phenotypes, and three distinct variants because of plectin mutations have been identified. First, EB with muscular dystrophy, an autosomal recessive syndrome, is frequently caused by premature termination codon causing mutations leading to the absence of plectin both in the skin and in the muscle. Second, a heterozygous missense mutation (R2110W) in PLEC1 has been documented in patients with EB simplex of the Ogna type, a rare autosomal dominant disorder. Finally, recent studies have disclosed plectin mutations in patients with EB with pyloric atresia, an autosomal recessive syndrome, frequently with lethal consequences. Collectively, these observations attest to the phenotypic spectrum of plectin mutations, and provide the basis for accurate genetic counselling with prognostic implications, as well as for prenatal diagnosis in families at the risk of recurrence of the disease. PMID- 15810883 TI - Early activation of cutaneous vessels and epithelial cells is characteristic of acute systemic onset juvenile idiopathic arthritis. AB - In biopsies of 16 patients (mean: 5.2 years) with acute systemic onset juvenile idiopathic arthritis (SOJIA), we analysed the initial cellular events during the characteristic cutaneous rash for composition of the infiltrate and for expression of activation markers on epithelial and endothelial cells. Despite the fleeting nature of the rash, there was a characteristic infiltration of neutrophils and monocytes, accompanied by a marked expression of endothelial adhesion receptors. In addition, we found a general activation of the cutaneous epithelium reflected by the expression of the pro-inflammatory S100-proteins - myeloid-related protein 8 (MRP8) and MRP14. In responders to therapy, follow-up biopsies showed a complete normalization of these inflammatory parameters, whereas non-responders presented with continuous signs of activation. In conjunction with the high level of epithelial activation, we detected an infiltrate of leucocytes within epithelium of sweat gland ducts during active SOJIA. Such a pattern has not been described for other inflammatory skin diseases nor did we find it in biopsies from nine patients with acute urticaria. It was accompanied by exclusive expression of MRP8, but not MRP14 by the secretory cells of sweat glands. Because MRP8 and MRP14, released by epithelial cells, exhibit pro-inflammatory effects on endothelial cells and leucocytes, the particular expression pattern of MRP8 and MRP14 in SOJIA is likely to represent a decisive early constitutive component in this inflammatory disease. Their differential expression further points to distinct roles of the individual molecules in inflammatory processes. PMID- 15810882 TI - CD1d and CD1d-restricted iNKT-cells play a pivotal role in contact hypersensitivity. AB - CD1d-restricted T-cells are activated by glycolipids presented by the major histocompatibility complex class-Ib molecule CD1d, found on the surface of antigen-presenting cells (APC). This interaction between APC, most notably dendritic cells (DC), and CD1d-restricted T-cells is an important regulatory step in the initiation of adaptive immune responses. It is well known that DC play a crucial role in the induction of contact hypersensitivity (CHS), a frequently studied form of in vivo T-cell-mediated immunity. In this study, we show that CD1d-restricted T-cells are also necessary for CHS, because both wild-type mice treated systemically or topically with CD1d glycolipid antagonists and CD1d restricted T-cell-null mice have markedly diminished CHS responses. Thus, pharmacologic antagonists of CD1d can be used as effective inhibitors of CHS, a prototype for a variety of delayed-type tissue hypersensitivity responses. PMID- 15810884 TI - Disruption of lipid rafts causes apoptotic cell death in HaCaT keratinocytes. AB - Lipid rafts are cholesterol-enriched microdomains in plasma membranes. The functional activity of many membrane proteins, including death and growth factor receptors, depends on their insertion in lipid rafts. We have previously demonstrated the presence of lipid rafts in keratinocytes and shown that lipid rafts are involved in the control of keratinocyte proliferation and metabolic activity. In this work, we investigated the effect of lipid-raft disruption on HaCaT keratinocyte survival. Lipid rafts could be disrupted or rearranged with cholesterol-targeting detergents: methyl-beta-cyclodextrin and filipin III. Moreover, cholesterol oxidation by a specific oxidase or blocking of cholesterol synthesis by mevastatin had a similar effect on lipid rafts. All cholesterol modifying substances caused cell death in a concentration-dependent manner. More detailed studies on the effects of cyclodextrin revealed apoptotic cell death at concentrations >or=0.5% (w/v). The molecular mechanism of apoptosis precipitated by raft disruption remains unknown but does not seem to be dependent of either membrane permeabilization or cell-cycle arrest imposed by cholesterol-modifying compounds. PMID- 15810885 TI - Limitations of CD95 ligand-transduced killer dendritic cells to prevent graft rejections. AB - As an attempt to experimentally induce antigen (Ag)-specific immunosuppression, we have previously created CD95 ligand (CD95L)-transduced dendritic cells (DC), which delivered apoptotic, but not activation, signals to CD4+ T cells in vitro in an Ag-dependent manner. We have also demonstrated that CD95L-transduced DC (termed killer DC) injected into syngeneic animals suppressed delayed-type hypersensitivity responses to an administered Ag. Based on these findings, we tested whether the injection of killer DC derived from A/J mice (H-2a) into allogeneic BALB/c recipients (H-2d) could prolong the survival of A/J-derived skin grafts by depleting A/J-reactive effector T cells. This attempt has not been successful. In this study, we elucidate the reasons for this failure, especially in terms of in vitro effects of killer DC on in vivo primed alloreactive T cells. We show that killer DC (i) failed to induce the proliferation of naive alloreactive T cells in a CD95/CD95L-dependent fashion, (ii) inhibited the proliferation of in vivo primed alloreactive T cells, (iii) killed relatively small fractions (up to 30%) of these T cells in vitro in a CD95/CD95L-dependent fashion and (iv) significantly, but incompletely, inhibited the generation of cytotoxic T-lymphocyte activities against A/J determinants. Thus, killer DC have significant, but modest, capacities to suppress in vitro alloimmune responses, which may not be sufficient to prolong the survival of alloskin grafts in a stringent allograft model. This study suggests that the current format of killer DC technology requires more modifications for its clinical application to prevent graft rejection. PMID- 15810886 TI - Identification and expression profiling of a human C-type lectin, structurally homologous to mouse dectin-2. AB - A number of C-type lectins on antigen-presenting cells play an important role in regulating innate immunity. Previously, we identified the mouse C-type lectins (dectin-1, and dectin-2) and human DECTIN-1. To identify human DECTIN-2, we employed degenerative polymerase chain reaction-based cDNA cloning using RNA from human Langerhans cell (LC)-like dendritic cells (DCs). This process yielded a cDNA encoding a C-type lectin with 66.5% amino acid sequence homology to mouse dectin-2, the same gene reported by Kanazawa et al. (J Invest Dermatol 2004: 122: 1522-1524) using the disparate approach of analyzing coding sequences in chromosome 12. Similar to their findings, we found gene expression in lung, spleen, and lymph node. Among resting leukocytes, it was expressed at highest levels by CD14+ monocytes, at lower levels by CD19+ B cells, and not at all by CD4+ T cells. Activation of CD19+ B cells with pokeweed mitogen down-regulated gene expression, whereas expression in CD4+ T cells was induced by Con A. Among our novel findings are an alternatively spliced transcript lacking exon 2, expression in bone marrow and tonsil, expression in CD8+ T cells that is abrogated following activation with phytohemagglutinin, restricted expression to CD1a+ LC within epidermis, and preferential expression by plasmacytoid (rather than myeloid) DC. Finally, we found that treatment with interleukin-4 (IL-4), IL 10, or UVB down regulated gene expression in CD14+ monocytes, whereas granulocyte macrophage colony-stimulating factor, transforming growth factor-beta1, or tumor necrosis factor-alpha treatment up-regulated it. Our findings may form the basis for understanding the function of human DECTIN-2 in innate immunity. PMID- 15810887 TI - Keratinocyte-specific modulation of type VII collagen gene expression by pro inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta). AB - Previous studies have shown that pro-inflammatory cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta up-regulate type VII collagen gene (COL7A1) expression in cultured dermal fibroblasts. The present study was designed to investigate the effects of TNF-alpha and IL-1beta on COL7A1 expression in epidermal keratinocytes. We demonstrated that both TNF-alpha and IL 1beta reduced COL7A1 expression in epidermal keratinocytes in an additive manner, whereas they increased COL7A1 expression in dermal fibroblasts. Thus, regulation of COL7A1 by pro-inflammatory cytokines is cell type specific. In particular, the inhibitory effects of TNF-alpha and IL-1beta occurred, at least in part, at the transcriptional level. Finally, we demonstrated that TNF-alpha and IL-1beta enhanced the TGF-beta-mediated up-regulation of COL7A1 expression in HaCaT keratinocytes, suggesting that the combination of TGF-beta and TNF-alpha or IL 1beta induces a signaling pathway that is completely different from that induced by either pro-inflammatory cytokine alone. PMID- 15810888 TI - Mast cell accumulation and cytokine expression in the tight skin mouse model of scleroderma. AB - The tight skin (Tsk) mouse develops many pathological changes seen in human scleroderma, such as increased collagen content and mast cell density. Although associations between mast cell expansion and skin fibrosis have been reported, the mechanisms underlying mast cell accumulation remain unclear. In this study, we have measured the density of skin mast cells in Tsk mice and their normal littermates (pa/pa) of 4-36 weeks of age, and in the skin heterografted between Tsk and pa/pa mice. Cytokines related to mast cell differentiation, proliferation and migration were examined by using RNase protection assays. Skin mast cell density in Tsk mice was significantly increased from 12 weeks of age, compared to that in pa/pa mice. The expression of transforming growth factor-beta1 (TGF beta1), and to a lesser extent, stem cell factor (SCF) and interleukin-15 (IL-15) mRNA was higher in Tsk mice, compared to that in control mice. Mast cell density was unchanged in Tsk skin grafted onto pa/pa hosts, but dramatically increased in pa/pa skin grafted onto Tsk hosts. This latter mast cell hyperplasia was associated with the increases in mRNA levels of TGF-beta1, SCF and IL-15, whereas little change in cytokine levels was seen in heterografted Tsk skin. These results suggest that locally produced cytokines in Tsk skin influence mast cell accumulation in this animal model of human scleroderma. PMID- 15810889 TI - Inhibition of monocyte-derived dendritic cell differentiation and interleukin-12 production by complement iC3b via a mitogen-activated protein kinase signalling pathway. AB - We have previously demonstrated that iC3b is deposited at the dermal-epidermal junction of the skin following ultraviolet (UV) exposure and that it plays a role in UV-induced immunosuppression and antigenic tolerance. In vitro, iC3b differentially regulates monocyte production of interleukin-10 (IL-10) and IL-12. Additionally, iC3b arrests monocytic cell differentiation into CD1c-expressing dendritic cell (DC) precursors. The present study addresses mitogen-activated protein kinase (MAPK) signalling following the cross-linking of CR3 by its ligand iC3b with regard to monocyte differentiation and cytokine regulation. Sheep erythrocytes were coated with IgM alone (EA) or iC3b (EAiC3b) to allow for CR3 cross-linking onto monocytes. EAiC3b increased the phosphorylation (p) of extracellular signal-regulated kinase (ERK) MAPK in fresh human monocyte, particularly in monocyte-derived DC (MDDC) that were differentiated by means of GM-CSF (1000 U/ml) and IL-4 (200 U/ml) for 2 days before iC3b exposure for an additional 24 h (P=0.034, n=3). CD1a expression, induced by GM-CSF and IL-4, was inhibited by iC3b (EAiC3b vs. EA, P=0.012, n=4). Conversely, the inhibition of ERK by the specific inhibitor (PD98059), but not the p-38 inhibitor SB203580, restored CD1a expression (P=0.011, n=4) in iC3b-stimulated MDDC. Concordantly, the inhibition of ERK during iC3b exposure fully reversed the inhibition of IL 12p70 induction in MDDC by 95% (P<0.01, n=4) and decreased IL-10 production. Taken together, our data demonstrate that iC3b interferes with MDDC differentiation and IL-12 and IL-10 production is mediated via an ERK MAPK dependent mechanism. Thus, ERK MAPK inhibition may represent a therapeutic strategy for preventing monocytic precursor diversion away from DC differentiation when monocytes enter injured tissues in which iC3b is generated, such as UV-exposed skin. PMID- 15810890 TI - Apparent fragility of African hair is unrelated to the cystine-rich protein distribution: a cytochemical electron microscopic study. AB - A feature of black African hair is an apparent increased fragility of the hair shaft compared to other ethnic groups (as measured by the tensile force needed to break the hair fibre). This has certain similarities to that reported for trichorrhexis nodosa (weathering secondary to physical damage) and trichothiodystrophy [a genetic disorder associated with reduced cystine (sulphur) rich proteins and increased fragility]. In the present study, the distribution of the cystine-rich proteins in the hair of black Africans was compared to that of Caucasian and Asian volunteers, plus patients with trichorrhexis nodosa and trichothiodystrophy, using transmission electron microscopy and specific silver stains. It was found that the silver staining pattern of the hair shafts of black Africans was similar to that observed for Caucasians, Asians and also patients with trichorrhexis nodosa. The cuticular cells exhibited an electron dense A layer and exocuticle, and in the cortex the microfibrils forming the macrofibres were outlined by electron-dense material. This contrasts with the abnormal distribution of the cystine-rich proteins seen in trichothiodystrophy. This study is the first formal comparison of the cystine-rich proteins in the various racial groups and shows that there is no abnormality in their distribution in black African hair shafts compared to the other ethnic groups. Therefore, the excessive structural damage observed in the African hair shafts is consistent with physical trauma (resulting from grooming) rather than an inherent weakness due to any structural abnormality. PMID- 15810891 TI - Nucleofection: a new, highly efficient transfection method for primary human keratinocytes*. AB - Transfection is an essential tool for numerous in vitro applications including studies of gene expression, promoter analysis, and intracellular signaling pathways and also for therapeutic strategies such as tissue engineering and gene therapy. However, transfection of primary cells including keratinocytes with common methods such as calcium phosphate, DEAE-dextran, liposome-mediated transfer, electroporation or viral vectors is problematic because of low transfection efficiency and the induction of terminal differentiation. Here we analyzed the use of nucleofection, a new, electroporation-based transfection method that enables the DNA to enter directly the nucleus, for the transfection of keratinocytes. Several different conditions were tested and optimized, resulting in a final transfection efficiency of 56% in primary human epidermal keratinocytes. This efficiency is superior to all non-viral transfection methods reported so far. The number of non-viable keratinocytes after nucleofection was low, varying between 14 and 16%. In contrast to other transfection protocols, nucleofection did not induce terminal differentiation in the transfected keratinocytes. In addition, nucleofection is a fast method, because the results can be analyzed within 7 h. In summary, nucleofection is a fast, easy and highly effective alternative for the transfection of primary human keratinocytes, which offers new opportunities for various research applications. PMID- 15810892 TI - New strategies in insulin treatment: analogues and noninvasive routes of administration. AB - Abstract Recent years have seen the development of alternatives to human insulin for the treatment of diabetes. Both rapid-acting and long-acting analogues are available. Alternative routes of insulin administration are emerging. The present review briefly summarizes the present knowledge on insulin analogues and alternative administration routes. PMID- 15810893 TI - Treatment of behavioural symptoms and dementia in Parkinson's disease. AB - Behavioural symptoms such as anxiety, depression and psychosis are common in Parkinson's disease (PD), and dementia occurs in about 90% of the patients. These symptoms can be more disabling than the motor dysfunction and they negatively impact quality of life, increase caregiver distress and are more frequently associated with nursing home placement. Depression can be treated with counselling and pharmacotherapy. Tricyclic antidepressants or selective serotonin reuptake inhibitors are widely used, but there is still need for controlled clinical trials. Management of psychosis in PD is complex and includes elimination of identifiable risk factors, reduction of polypharmacy and administration of atypical neuroleptics, which can alleviate psychotic symptoms without worsening motor functions. Clozapine is the best documented atypical neuroleptic shown to be effective against psychosis in PD patients. Cholinesterase inhibitors may prove additional benefit in psychotic PD patients. Recent evidence from small double-blind and open-label trials suggests that cholinesterase inhibitors may be effective in the treatment of dementia associated with PD. PMID- 15810894 TI - A search for presynaptic beta3-adrenoceptors in the rat. AB - Presynaptically localized adrenoceptors occur on a variety of neurones. In particular, alpha2-adrenoceptors, occurring on neurones of the peripheral and central nervous system, inhibit the release of the respective transmitters whereas beta2-adrenoceptors on some types of postganglionic sympathetic neurones facilitate noradrenaline release. Since only little information is available whether there are also presynaptic beta3-adrenoceptors, we examined the effect of beta3-adrenoceptor agonists on noradrenaline release from the resistance vessels and the hippocampus of the rat and on serotonin and acetylcholine release from the rat hippocampus. In rat hippocampal slices preincubated with (H noradrenaline, 3H-serotonin and 3H-choline and superfused in the presence of an inhibitor of the neuronal transporter of the respective neurone, the beta3 adrenoceptor agonist CL 316243 did not affect the electrically evoked tritium overflow. The latter was, however, inhibited by at least 50% by agonists of the respective autoreceptors. CL 316243 and another three beta3-adrenoceptor agonists (BRL 37344, ZD 2079 and CGP 12177) failed to affect the electrically evoked tritium overflow also in slices preincubated with 3H-noradrenaline and superfused in the presence of the alpha2-adrenoceptor antagonist rauwolscine whereas prostaglandin E2 caused a marked inhibition. In pithed and vagotomized rats, the increase in diastolic blood pressure induced by electrical stimulation of the sympathetic outflow was also not affected by CL 316243 but markedly inhibited by the cannabinoid receptor agonist WIN 55212-2. CL 316243 and WIN 55212-2 were devoid of an effect on the rise in diastolic blood pressure induced by exogenous noradrenaline. In conclusion, our data suggest that the noradrenergic neurones innervating the resistance vessels of the rat and the noradrenergic, serotoninergic and cholinergic neurones of the rat hippocampus are not endowed with presynaptic beta3-adrenoceptors. PMID- 15810895 TI - Taurine treatment protects against chronic nicotine-induced oxidative changes. AB - Experiments have shown that chronic nicotine administration caused oxidative damage in various organs by increasing lipid peroxidation products and decreasing the activity of endogenous antioxidants. The aim of this study was to investigate the effects of taurine treatment on nicotine-induced oxidative changes in rat thoracic aorta and heart and to explore the possible mechanisms of action. Male Wistar albino rats (200-250 g) were injected with nicotine hydrogen bitartrate (0.6 mg/kg; i.p.) or saline for 21 days. Taurine was administered (50 mg/kg; i.p.) alone or along with nicotine injections. After decapitation, the thoracic aorta and heart tissues were excised. The aorta was used for in vitro contractility studies or stored along with the heart samples for the measurement of malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and collagen content. Tissue samples were also examined histologically. Serum samples were stored for the measurement of MDA, GSH and lactate dehydrogenase (LDH) activity. Chronic nicotine treatment impaired both the contraction and relaxation responses of the aortic rings to phenylephrine and acetylcholine, respectively. It increased lipid peroxidation, MPO levels and tissue collagen content of both aorta and heart samples. Taurine supplementation to nicotine-treated animals reversed the contractile dysfunction and restored the endogenous GSH levels and decreased high lipid peroxidation and MPO activities in both tissues. These data suggest that taurine supplementation effectively attenuates the oxidative damage because of chronic nicotine administration possibly by its antioxidant effects. PMID- 15810896 TI - Acute cardiodepressant effects induced by bolus intravenous administration of amiodarone in rabbits. AB - Amiodarone is a potent anti-arrhythmic with a large pharmacological spectrum that shares the mechanisms of action of all classes of anti-arrhythmic drugs. Originally used in the treatment of supraventricular arrhythmias, it has also been used to treat ventricular tachyarrhythmias. The recent inclusion of amiodarone in the Advanced Cardiac Life Support protocols warrants the characterization of the hemodynamic profile resulting from the rapid venous administration of the drug. Thus, the main purpose of the present study was to investigate the acute hemodynamic profile resulting from the bolus i.v. injection of amiodarone, compared with bolus i.v. administration of lidocaine. We investigated the acute hemodynamic effects of amiodarone and lidocaine, in an experimental model of open-chest pentobarbital-anesthetized rabbits (n = 24). Amiodarone (5 mg/kg) induced immediate reductions in mean arterial pressure (MAP) of 32 +/- 5% (P < 0.001), accompanied by reductions in cardiac contractility and relaxation, as assessed by left ventricular (LV) +dP/dt(max) and -dP/dt(max) (40 +/- 4 and 36 +/- 4% respectively) (P < 0.001), heart rate (HR) 10 +/- 1% (P < 0.05), cardiac output (CO) 24 +/- 5% (P < 0.001) and systemic vascular resistance (SVR) 19 +/- 3.5% (P < 0.05). Lidocaine (3 mg/kg) induced reductions in: MAP of 18 +/- 7% (P < 0.001), LV +dP/dt(max) and -dP/dt(max) (40 +/- 5 and 22 +/- 7% respectively) (P < 0.001), HR 7 +/- 1% (P < 0.01) and CO of 23 +/- 6% (P < 0.001). SVR increased by 9 +/- 1.5% (P > 0.05). It is concluded that rapid i.v. administration of both amiodarone and lidocaine induces significant cardiovascular depression mainly characterized by immediate reductions in cardiac contractility. PMID- 15810897 TI - Gentamicin-induced nephrotoxicity and protective effect of caffeic acid phenethyl ester in rats. AB - The objective of this study was to investigate the beneficial effects of caffeic acid phenethyl ester (CAPE) on gentamicin (GM)-induced nephrotoxicity in Wistar rats. Twenty-one adult Wistar rats were divided into three groups as follows: control group, GM and GM + CAPE group. Control group rats were injected with 5% ethanol, GM group rats were treated with 100 mg/kg GM and GM + CAPE group were pretreated with 10 mumol/kg CAPE for 2 days, then exposed to GM at the same dose. Drug injections were applied for 12 days. Twenty-four hours after the last injection, rats were killed and kidneys were quickly removed. Tissue malondialdehyde (MDA) measurements and microscopic examination of kidneys were performed. In the GM group, significant increases in MDA levels were observed (P < 0.05). These changes were found to be normalized in the GM + CAPE group. Exposure to GM caused necrosis of tubular epithelial cells. Necrosis of tubules were found to be prevented by CAPE pretreatment. In conclusion, CAPE exerted an improvement on GM-induced nephrotoxicity, possibly, at least in part through inhibition of the production of oxygen free radicals that cause lipid peroxidation. PMID- 15810898 TI - Paradoxal effect of salbutamol in an in vitro model of bronchoprotection. AB - Salbutamol-induced hyperresponsiveness to acetylcholine was investigated in human and guinea-pig isolated airways and cultured human airway smooth muscle cells. Salbutamol (10(-7)-10(-5) m) inhibited contractions induced by low concentrations of acetylcholine (10(-8)-10(-7) m) but potentiated contractions induced by higher concentrations of acetylcholine (10(-5)-10(-3) m). Pretreatment with the calcium channel antagonist nicardipine suppressed salbutamol-induced hyperresponse. Stimulation of cultured human airway smooth muscle cells with salbutamol (10(-6) m) amplified intracellular calcium concentration rise induced by acetylcholine (10(-5) m). Propranolol (10(-7) m), a beta1- and beta2-adrenoceptor antagonist, and ICI 118551 (10(-7)-10(-6) m), a beta2-adrenoceptor antagonist, suppressed the inhibitory effect of salbutamol but did not inhibit the hyperresponse on high concentrations of acetylcholine. In contrast, higher concentration of propranolol (10(-6) m) inhibited salbutamol-induced hyperreactivity. Effects of salbutamol were not affected by atenolol, a beta1-adrenoceptor blocker. Salbutamol-induced hyperresponsiveness is mediated through a mechanism involving calcium channel activation. PMID- 15810899 TI - Efficacy and renal tolerability of ibuprofen vs. indomethacin in preterm infants with patent ductus arteriosus. AB - Indomethacin is commonly used for the treatment of patent ductus arteriosus (PDA) but has renal failure as a main side-effect. Ibuprofen seems to be efficient in closing the ductus with less side-effects, but few studies are available in literature as regards its use in preterm infants. This study is a retrospective analysis of clinical data in order to compare the efficacy and the renal tolerability of ibuprofen and indomethacin administered to preterm infants with gestational age (GA) < or = 30 weeks for the treatment of PDA. From our data, ibuprofen results pharmacologically as efficient as indomethacin and could be an alternative in prematures. About renal tolerability, our data confirm that non steroidal anti-inflammatory drugs treatment could affect at least transiently renal function. Moreover, indomethacin could be more nephrotoxic compared with ibuprofen, as creatinine concentrations normalize more slowly in this group, although the mean difference between the two drugs was not significant as our population sample was small. Further studies are needed to assess whether ibuprofen is really less nephrotoxic than indomethacin, in particular by examining carefully the correlation between GA and ibuprofen administration. PMID- 15810900 TI - Abstracts of the 9th Annual meeting of the Societe Francaise de Pharmacologie, 26th Pharmacovigilance meeting and 11th CHU CIC meeting. Bordeaux 26-28 April 2005. PMID- 15810901 TI - The clinical syndrome of Alzheimer's disease: aspects particularly relevant to clinical trials. AB - This paper describes the natural history of the clinical syndrome of Alzheimer's disease (AD) including the cognitive deficit, the neuropsychiatric symptoms, impact on daily functioning, risk factors, medical complications and impact on the use of health-care resources. The clinical presentation of the disease varies greatly from the prodrome through end stage; instruments used to quantify the severity of each aspect of the disease have been developed and are described along with their use in clinical drug trials. Drug treatments for AD are usually developed by first showing a positive effect on the cognitive deficit, with later studies investigating drug effects on other clinical aspects of the disease. PMID- 15810902 TI - Genetic and molecular aspects of Alzheimer's disease shed light on new mechanisms of transcriptional regulation. AB - Rapid advances made in biological research aimed at understanding the molecular basis of the pathogenesis of Alzheimer's disease have led to the characterization of a novel catalytic activity termed gamma-secretase. First described for its beta-amyloid-producing function, gamma-secretase is now actively studied for its role in a novel signal transduction paradigm, which implicates cell-surface receptor proteolysis and direct surface-to-nucleus signal transduction. gamma Secretase targets numerous type I protein receptors involved in diverse functions ranging from normal development to neurodegeneration. In this Review we discuss how the study of the genetic and molecular aspects of Alzheimer's disease has revealed a dual role of gamma-secretase in transcriptional regulation and in the pathogenesis of familial Alzheimer's disease. PMID- 15810903 TI - Invertebrate models of Alzheimer's disease. AB - The intensely studied model organisms Caenorhabditis elegans and Drosophila melanogaster have been employed to study a number of neurodegenerative diseases, including Alzheimer's disease (AD). Although worms and flies are phylogenetically distant from humans, results of both classic genetic analyses and transgenic manipulation of these invertebrates suggest they are valid models for at least some aspects of AD. This review describes the rationale for AD-relevant studies in worms and flies and discusses both what has been learned from these studies and what may be discovered in the future. PMID- 15810904 TI - A case for a non-transgenic animal model of Alzheimer's disease. AB - Alzheimer's disease (AD) is associated with an early impairment in memory and is the major cause of dementia in the elderly. beta-Amyloid (Abeta) is believed to be a primary factor in the pathogenic pathway leading to dementia. Mounting evidence suggests that this syndrome begins with subtle alterations in synaptic efficacy prior to extensive neuronal degeneration and that the synaptic dysfunction could be caused by diffusible oligomeric assemblies of Abeta. This paper reviews the findings from behavioral analysis, electrophysiology, neuropathology and nootropic drug screening studies involving exogenous administration of Abeta in normal rodent brains. This non-transgenic model of amyloid pathology in vivo is presented as a complementary alternative model to transgenic mice to study the cellular and molecular pathways induced by amyloid, which in turn may be a causal factor in the disruption of cognition. The data reviewed here confirm that the diffusible form of Abeta rapidly induces synaptic dysfunction and a secondary process involving cellular cascades induced by the fibrillar form of amyloid. The time-course of alteration in memory processes implicates at least two different mechanisms that may be targeted with selective therapies aimed at improving memory in some AD patients. PMID- 15810905 TI - Behavioral phenotypes of amyloid-based genetically modified mouse models of Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common neurodegenerative affliction of the elderly, presenting with progressive memory loss and dementia and terminating with death. There have been significant advances in understanding the biology and subsequent diagnosis of AD; however, the furious pace of research has not yet translated into a disease-modifying treatment. While scientific inquiry in AD is largely centered on identifying biological players and pathological mechanisms, the day-to-day realities of AD patients and their caregivers revolve around their steady and heartbreaking cognitive decline. In the past decade, AD research has been fundamentally transformed by the development of genetically modified animal models of amyloid-driven neurodegeneration. These important in vivo models not only replicate some of the hallmark pathology of the disease, such as plaque-like amyloid accumulations and astrocytic inflammation, but also some of the cognitive impairments relevant to AD. In this article, we will provide a detailed review of the behavioral and cognitive deficits present in several transgenic mouse models of AD and discuss their functional changes in response to experimental treatments. PMID- 15810906 TI - Working memory impairment in a transgenic amyloid precursor protein TgCRND8 mouse model of Alzheimer's disease. AB - The most profound deficits observed in Alzheimer's disease (AD) are in domains of episodic and working memory systems. Transgenic (Tg) mice expressing mutated human amyloid precursor protein (APP) genes offer a model to study the effect of AD pathology on cognition. We reported previously that APP TgCRND8 mice showed deficits in a reference and working memory evaluated in a Morris water-maze test. In this study, we evaluated the working memory of TgCRND8 mice comparing two training paradigms in a six-arm radial water maze. In the first paradigm, the exploration of the maze was constrained, forcing the mice to use a spatial mapping strategy. In the second paradigm, mice were unconstrained in their exploration of the maze. TgCRND8 mice proved to be significantly impaired in spatial working memory in both paradigms as compared with their non-transgenic littermates. The analysis of data revealed that forcing mice to use a spatial strategy during training caused only a moderate improvement in the performance of all mice. However, unconstrained exploration of the maze not only resulted in a fast learning in control mice, but also facilitated the development of a chaining strategy in spatially impaired TgCRND8 mice. In conclusion, TgCRND8 mice showed impairment in spatial working memory but retained a plasticity to choose alternative search strategies. PMID- 15810907 TI - Prophylactic treatment effects on inhibitor risk: experience in one centre. AB - Nowadays, the elective treatment for children with haemophilia is prophylaxis. There is a common consensus that this modality of therapeutic approach is not associated with a higher risk of inhibitor development. We analysed the inhibitor incidence in 50 haemophiliac children and its relationship with mutations, type of clotting factor used and treatment modality. There was a significant correlation between receiving on-demand treatment and an increased incidence of inhibitors, independently of mutations or factor used. We advise putting haemophiliac children under prophylactic treatment as soon as possible, especially if they have mutations associated with high risk of inhibitor development, as prophylaxis is negatively associated with the development of inhibitors. PMID- 15810908 TI - B-domain deleted recombinant factor VIII preparations are bioequivalent to a monoclonal antibody purified plasma-derived factor VIII concentrate: a randomized, three-way crossover study. AB - BACKGROUND: Deletion of the B-domain of recombinant blood coagulation factor VIII (BDDrFVIII) increases the manufacturing yield of the product but does not impair in vitro or in vivo functionality. BDDrFVIII (ReFacto) has been developed with the additional benefit of being formulated without human albumin. OBJECTIVE: The primary objective of this three-way crossover-design study was to compare the pharmacokinetic (PK) parameters of two BDDrFVIII formulations (one reconstituted with 5 mL of sterile water, the other reconstituted with 4 mL sodium chloride 0.9% USP) with those of a plasma-derived, full-length FVIII preparation (Hemofil M) in patients with haemophilia A to determine bioequivalence. METHODS: A series of blood samples were collected over a period of 48 h after i.v. administration of each of the FVIII preparations. Plasma FVIII activity was determined using a validated chromogenic substrate assay. Plasma FVIII activity vs. time curves was characterized for a standard set of PK parameter estimates. Two parameter estimates, the maximum plasma concentration (Cmax) and the area under plasma concentration vs. time curves (AUCs), were used to evaluate bioequivalence. The two preparations were considered bioequivalent if the 90% confidence intervals for the ratio of geometric means for Cmax and AUCs fell within the bioequivalence window of 80% to 125%. RESULTS/CONCLUSION: Results show that each BDDrFVIII formulation is bioequivalent to Hemofil M and the two formulations of BDDrFVIII are bioequivalent to each other. PMID- 15810909 TI - Changing pattern of care of boys with haemophilia in western European centres. AB - Haemophilia management is not uniform among countries, even within western Europe, that have close economic, social and cultural relationship. The European Paediatric Network PedNet aims to share experiences in the field of the care of boys with haemophilia. In 1998, a PedNet survey has shown significant disparities in 20 centres from 16 countries, particularly as regards the implementation of prophylaxis regimen. This survey has been updated in 2003 to describe the current status of haemophilia management in 22 centres and the changing pattern of care of boys with severe haemophilia in western Europe. Regular, continuous long-term prophylaxis is provided in all PedNet centres, more than 50% and 80-100% of boys being treated this way in 20/22 and 15/22 centres respectively. Twenty of the 22 centres (91%) recommend continuous prophylaxis (primary or secondary A) for a new patient. The use of recombinant factor VIII concentrates was already widespread in 1998 and a further expansion of recombinant products has been observed over the last 5 years. Recombinant FVIII is now used exclusively in nine centres and for more than 80% of boys with haemophilia A in nine other centres. The use of recombinant and plasma derived FIX is more balanced: among 18 centres where boys with haemophilia B are treated, 14 use recombinant FIX, and nine administer it to a majority of patients. Other modifications of practice have been stressed in this survey, such as more targeted use of central venous devices in the youngest boys and more extensive characterisation of genetic mutations. PMID- 15810910 TI - Dose effect and efficacy of rFVIIa in the treatment of haemophilia patients with inhibitors: analysis from the Hemophilia and Thrombosis Research Society Registry. AB - Recombinant activated factor VII (rFVIIa), licensed in 1999 for treatment of haemophilia patients with inhibitors (HI), represents an important advance in the therapeutic armamentarium. Standard bolus dosing ranges from 90 to 120 mcg kg(-1) every 2-3 h until arrest of bleeding. As licensure, clinical use of rFVIIa has increased and broadened. Clinicians now use a wide dose range, 90-300 mcg kg(-1). High-dose regimens may optimize thrombin generation or burst, and may allow for prolonged dose interval. The Hemophilia and Thrombosis Research Society (HTRS) maintains a registry database to study haemophilia treatment and related disorders, particularly treatment of acute bleeding in HI, acquired haemophilia, FVII deficiency and von Willebrand's disease (VWD). To assess the effect of rFVIIa dose on efficacy and safety in the treatment of acute bleeding in HI, data from the HTRS database from January 2000 through June 2002 were analysed. Bleeding episodes were grouped by bolus rFVIIa dose range: <100, 100-150, 150-200 and >200 mcg kg(-1). Investigator-reported efficacy for the first 72 h of treatment was evaluated. Thirty-eight congenital HI patients were treated for 555 bleeding episodes. Patient age range was 1-55 years (median: 14). Bleeding episodes were spontaneous (45%), caused by trauma (38%), or because of surgery, dental, diagnostic, or medical procedures (17%); bleeding occurred in joint, muscle, and intra/extracranial sites. Treatment location included: 80% at home, 12% at other facilities (treatment centres, ER, inpatient and OR), and 8% at both home/other facilities. Median total dose given over 72 h was 360 mcg kg(-1) (range: 40-4281, mean: 537). Bleeding stopped in 87% of the episodes. Bleeding cessation rate was 84% for the three lower dose groups, and 97% for the highest dose group (P < 0.001). Five patients experienced nine adverse events (AEs). AE rates were <1% for <100, 5% for 100-150, 0% for 150-200, <1% for >200 mcg kg(-1) dose group. Decreased therapeutic response accounted for eight of the nine AEs. These data, which represent the most comprehensive report of rFVIIa use since the USA licensure, demonstrate that bleeding episodes in HI patients can be treated safely and effectively at home and that doses up to 346 mcg kg(-1) appear to be well-tolerated. Additionally, rFVIIa doses >200 mcg kg(-1) appear to significantly increase efficacy (97% in the high-dose group, compared with 84% in the lower dose groups). Optimal dosing remains to be determined; specifically, what the lowest effective dose is and whether a single high-dose bolus eliminates the need for repeated dosing. Recombinant FVIIa appears to have a wide safety margin that may allow dose escalation to address these questions. PMID- 15810911 TI - Commentary: magnetic resonance imaging as an evaluative tool in haemophilic arthropathy, on behalf of the Medical Advisory Board, World Federation of Haemophilia and the International Prophylaxis Study Group. PMID- 15810912 TI - Compatible scales for progressive and additive MRI assessments of haemophilic arthropathy. AB - The international MRI expert subgroup of the International Prophylaxis Study Group (IPSG) has developed a consensus for magnetic resonance imaging (MRI) scales for assessment of haemophilic arthropathy. A MRI scoring scheme including a 10 step progressive scale and a 20 step additive scale with identical definitions of mutual steps is presented. Using the progressive scale, effusion/haemarthrosis can correspond to progressive scores of 1, 2, or 3, and synovial hypertrophy and/or haemosiderin deposition to 4, 5, or 6. The progressive score can be 7 or 8 if there are subchondral cysts and/or surface erosions, and it is 9 or 10 if there is loss of cartilage. Using the additive scale, synovial hypertrophy contributes 1-3 points to the additive score and haemosiderin deposition contributes 1 point. For osteochondral changes, 16 statements are evaluated as to whether they are true or false, and each true statement contributes 1 point to the additive score. The use of these two compatible scales for progressive and additive MRI assessments can facilitate international comparison of data and enhance the accumulation of experience on MRI scoring of haemophilic arthropathy. PMID- 15810913 TI - MRI scores of ankle joints in children with haemophilia--comparison with clinical data. AB - Fifty-six ankle joints in 38 haemophilic boys were investigated by magnetic resonance imaging (MRI) and the findings were classified according to both the Denver- and the European scoring schemes. The different MRI scores were compared with each other and with clinical data on number of joint bleeds and the orthopaedic joint score. MRI changes that were more advanced than a small effusion were found in 33 (59%) of the ankles and osteochondral changes were observed in 20 (36%). The total number of bleeds in individual ankles ranged from 0 to 80 (mean: 11). The orthopaedic joint score was 0 for 49 ankles and ranged from one to four for seven ankles. There was a significant and strong correlation between the assessment results obtained with the two MRI scoring methods (correlation coefficients ranged from 0.80 to 0.95, P < 0.001), and both types of MRI scores were weakly but significantly correlated with the clinical data on the number of joint bleeds and the orthopaedic joint score (correlation coefficients: 0.32-0.39, P < 0.01 or P < 0.05). MRI is useful for evaluating early joint changes in patients with haemophilia. The European scoring method differentiates the arthropathic changes further than the Denver scale does, but the two different scoring methods have similar correlation to the number of joint bleeds and the orthopaedic joint score. PMID- 15810914 TI - An exploration of health-related quality of life in adults with haemophilia--a qualitative perspective. AB - Musculoskeletal dysfunction is a common feature of haemophilia and along with other manifestations of this condition, there is a general perception that health related quality of life (QoL) will be affected. Previous research using standardized questionnaires has demonstrated that QoL is lower in haemophilia groups compared with normal populations. However, disability studies and interviews with disabled people suggest that many disabled people experience positive life changes as a result of their illness and an affirmative model of disability has been proposed. A qualitative study involving focus groups and interviews was undertaken to explore these issues in a group of 19 severely affected adults with haemophilia. The focus groups and interviews were tape recorded and fully transcribed and the results subjected to thematic analysis. This paper focuses specifically on key issues that impacted on perceptions of QoL. The findings suggest that the participants' perceptions of their QoL were very positive. Possible reasons for this are proposed including the benefits of factor replacement, participants' recollections of their lifestyle before factor replacement, that having haemophilia was integral to the 'self' and finding a niche where they could be successful. A more positive affirmation of haemophilia may need to be considered when developing QoL measures. PMID- 15810915 TI - Mutation analysis in 51 patients with haemophilia A: report of 10 novel mutations and correlations between genotype and clinical phenotype. AB - We report the results of genetic analysis on a series of 51 patients attending this Haemophilia Comprehensive Care Centre. The most common cause of severe haemophilia A--the factor VIII intron 22 inversion was detected in eight families and the factor VIII intron 1 inversion in three families. Mutation analysis was carried out on the remaining patients by nucleotide sequencing of genomic DNA after screening with conformation-sensitive gel electrophoresis (CSGE) or denaturing high-performance liquid chromatography (dHPLC). A total of 27 different FVIII non-inversion mutations were detected. Severe haemophilia was associated with 12 null mutations (six nonsense, six frameshift) and four missense mutations. A further 11 different missense mutations were associated with moderate or mild disease. To our knowledge, six null mutations [1950del 4(tttg), 3270-75insA, 4416del 10, 6735-38delA, W1029X, Y1792X] and four missense mutations (E1682K, M1947V, P2048L, P2143L) have not been previously published. Each novel missense mutation occurred at a highly conserved residue, no other candidate mutation was detected on screening the entire coding region of the FVIII gene and they were not detected in a screen of individuals without haemophilia A. The genotype-phenotype correlations of the FVIII mutations detected will be discussed. PMID- 15810916 TI - Utility of a (GT) dinucleotide repeat in intron 1 of the factor 8 gene for haemophilia A carrier diagnosis. AB - We report the usefulness of a dinucleotide GT repeat in intron 1 of the factor 8 gene in carrier diagnosis of haemophilia A (HA). We analyzed 47 women from HA families in which the mutation was not identified in the index case and in which the common intragenic polymorphic markers were uninformative. The intron 1 GT repeat was useful to identify the X chromosome with the mutation in 19 of them. The analysis of this easily detectable marker in conjunction with other current markers may facilitate X chromosome identification in a large proportion of HA families. PMID- 15810917 TI - A framework for genetic service provision for haemophilia and other inherited bleeding disorders. AB - This framework document offers guidance to patients, doctors, nurses, laboratory scientists, funders and hospitals on the provision of clinical and laboratory genetic services for haemophilia. With recent advances in molecular laboratory techniques it is now possible to give the vast majority of individual patients and family members very reliable genetic information. To enable these genetic data to be used for both the optimal treatment of patients with inherited bleeding disorders and for appropriate reproductive decisions in carriers, there needs to be a clear and robust framework for systematically acquiring the necessary clinical, personal, family and laboratory information upon which decisions can be made. This document provides guidance on the range and standards of clinical and laboratory genetic services which should be offered to patients and their families. Included are arrangements for genetic counselling and testing (including consent and confidentially issues), management of early pregnancy, standards for laboratory genetic services, as well as advice on data storage, security and retrieval. PMID- 15810918 TI - Acquired factor VIII inhibitor presenting as macular haemorrhage. AB - We report a rare case of idiopathic acquired factor VIII inhibitor in an 80-year old Chinese man presented as sudden onset of monocular blindness because of macular haemorrhage. This was complicated by painful glaucoma that did not respond to medical treatment. The patient died of cerebral haemorrhage shortly afterwards. Most cases of reported intraocular bleeding in acquired haemophiliacs are iatrogenic because of intraocular operations in undiagnosed cases, and spontaneous intraocular haemorrhage has never been reported. The literature reports of intraocular bleeding in hereditary and acquired haemophilia cases are summarized. PMID- 15810919 TI - The use of activated recombinant coagulation factor VII during haemarthroses and synovectomy in a patient with congenital severe factor V deficiency. AB - Factor V deficiency is a rare hereditary bleeding disorder. Currently, FV concentrates are not available, and the treatment of spontaneous bleeding or bleeding associated with invasive procedures is transfusion of fresh frozen plasma (FFP). However, FFP transfusion can lead to the development of inhibitor to FV, and is associated with several potential transfusion reactions including allergic reactions. We report a patient with congenital severe FV deficiency with repeated haemarthroses of a shoulder joint, and progressively severe allergic reactions to FFP transfusions. In addition, the patient also developed acute pulmonary oedema. Activated recombinant coagulation factor VII (rFVIIa) was used as an alternative haemostatic agent to FFP. We describe the use of rFVIIa in this patient during haemarthroses, synovectomy, and physiotherapy. PMID- 15810920 TI - The use of recombinant factor VIIa for bleeding in paediatric practice. AB - Summary. Recombinant factor VIIa is licensed for use in patients with haemophilia and inhibitors. More recently it has been used as an unlicensed product in adults for the treatment of life threatening bleeding. However, its use in paediatric practice has been limited. We describe a series of patients from one paediatric centre, where recombinant factor VIIa has been used when conventional treatment has failed to achieve haemostasis. PMID- 15810921 TI - Orthopaedic manifestations of the hip in haemophilia--impaction bone grafting used in revision total hip arthroplasty in a haemophilic patient. AB - The condition of haemophilia presents many challenges to the orthopaedic surgeon. Two such challenges include the haemophilic pseudotumour, a rare complication occurring in 1-2% of haemophiliacs, and the phenomenon of aseptic loosening of arthroplasty implants. This report presents a case involving the separate management of a pseudotumour and revision hip arthroplasty in the same hip using impaction bone grafting. PMID- 15810922 TI - A case of an asymptomatic idiopathic inhibitor to coagulation factor V. PMID- 15810923 TI - Acquired haemophilia in a patient with gram-negative urosepsis and bladder cancer. AB - We here report a patient who developed a high titer antibody to factor VIII (FVIII) during gram-negative urosepsis caused by enterobacter cloacae after complete resection of rectal cancer. The patient initially presented with a life threatening spontaneous hematothorax and multiple large haematomas. Coagulation studies revealed a severe FVIII deficiency <1% with a high FVIII antibody titer of 64 BU. The bleeding responded rapidly to infusions of recombinant factor VIIa. After achievement of a partial remission (FVIII activity 28%) by combined immunosuppressive therapy (prednisone, cyclophosphamide, plasmapheresis and immunoadsorption), subsequently, two relapses occurred following steroid tapering. Resumption of prednisone and cyclophosphamide treatment combined with immunoadsorption induced a second and third remission, respectively. After resection of a papillary carcinoma of the bladder 6 months later and continuous immunosuppressive therapy with cyclophosphamide, FVIII levels remained stable within normal ranges. This clinical course suggests that the cause of inhibitor formation against FVIII resulting in severe acquired haemophilia was multifactorial and was initiated by the gram-negative urosepsis and probably by the underlying malignancies. PMID- 15810924 TI - Management of neonates with inherited bleeding disorders--a survey of current UK practice. PMID- 15810925 TI - Pathological fracture of a true tumour mimicking a haemophilic pseudotumor. PMID- 15810926 TI - Synoviorthesis in haemophilic synovitis: which is the best radioactive material to use? PMID- 15810927 TI - A little knowledge is a dangerous thing ... PMID- 15810928 TI - The value and impact of information provided through library services for patient care: a systematic review. AB - OBJECTIVE: An updated systematic review was carried out of research studies looking at the value and impact of library services on health outcomes for patients and time saved by health professionals. METHODS: A comprehensive systematic search was undertaken of the published literature to September 2003 in ERIC, LISA, MEDLINE, PREMEDLINE, EMBASE, the Cochrane Controlled Trials Register and Google. Some handsearching was carried out, reference lists were scanned and experts in the field were contacted. Twenty-eight research studies of professionally led libraries for health-care staff, including clinical librarian projects, met the inclusion criterion of at least one health or 'time saved' outcome. Papers were critically appraised using internationally accepted criteria. Data were extracted and results were summarised using a narrative format as the studies were heterogeneous and precluded a statistical analysis. RESULTS: There is evidence of impact from both traditional and clinical librarian services. The higher quality studies of traditional services measured impacts of 37-97% on general patient care, 10-31% on diagnosis, 20-51% on choice of tests, 27-45% on choice of therapy and 10-19% on reduced length of stay. Four studies of clinical librarian projects suggested that professionals saved time as a result of clinical librarian input, and two of these studies showed evidence of cost effectiveness. However, the clinical librarian studies were generally smaller, with poorer quality standards. CONCLUSIONS: Research studies suggest that professionally led library services have an impact on health outcomes for patients and may lead to time savings for health-care professionals. The available studies vary greatly in quality but the better quality studies also suggest positive impacts. Good practice can be gathered from these studies to guide the development of a pragmatic survey for library services that includes the direct effects for patients among the outcome measures. PMID- 15810929 TI - A survey of UK clinical librarianship: February 2004. AB - AIM: This article will describe a survey carried out in February 2004, the aim of which was to summarize the form and content of clinical librarian (CL) and other similar outreach information services to UK health professionals in the acute (secondary or tertiary) sector. OBJECTIVES: (i) To survey the activities and views of UK information professionals offering information services involving the librarians' presence in the clinical setting, (ii) to develop a tool to explore critical aspects of this form of information work, (iii) to create a contacts database for UK CLs, to be made available on the Internet. METHODOLOGY: All known information specialists/librarians offering CL or similar services were surveyed. The semi-structured questionnaire was piloted. Respondents were asked to consider their activity over a period of 4 weeks. RESULTS: Twenty-six people responded to the invitation to take part and met the inclusion criteria. CONCLUSIONS: A summary of a 'typical' clinical librarian revealed by this survey is given, with a major conclusion that there is a very mixed picture of activity. Opinion on how far CLs should go in fully appraising search results is uncertain. The survey suggests reasons for this and the developments that may influence change are discussed. Recommendations for future research and development are offered. PMID- 15810930 TI - The information needs of perioperative staff: a preparatory study for a proposed specialist library for theatres (NeLH). AB - AIMS/OBJECTIVES: To undertake an information-needs analysis of operating theatre staff in order to inform the development of a specialist library of the National Electronic Library of Health (NeLH). SETTING: Three hospitals in an NHS Trust in a rural county in the north of England. METHODS: In-depth interviews, during which staff were questioned in regard to their job roles and information needs, information-seeking behaviour, access to computers, use and perceptions of the NeLH, and what they would like from a specialist library. RESULTS: Information needs were found to concern direct care, professional and academic development, medical devices knowledge and administration. Currency and speed of delivery were the key information requirements. Poor communication appeared to be a major problem for many staff and inhibited efficient information exchange. When using the Web, both clinical and non-clinical staff searched in a similar fashion to health consumers generally, despite their need for highly specialist information. Most respondents reacted warmly to the idea of the proposed National Electronic Library for Theatre Staff (NeLTS) and made several practical suggestions for suitable material. However, access was problematic for nurses and might prove to be an obstacle to take-up of the Specialist Library. CONCLUSIONS: Communication difficulties, the fragmentation of data in the field and the consequent difficulties of finding information point to the need for an NeLTS. However, for this library to embrace and succeed with all the various groups that constitute the operating theatre staff, issues of access and digital literacy need to be addressed. PMID- 15810931 TI - Influence of strategic direction for NHS Scotland knowledge services on indexing policy for the NHS Scotland e-library. AB - OBJECTIVES: Indexing policy for the NHS Scotland e-Library needs to maximize future inter-operability with other significant health- and social-care-related resources. The strategic drive towards integration and partnership working means that the indexing system has to be widely acceptable to the full range of disciplines within the integrated health-care family. METHODS: Indexes identified by various means and then shortlisted using predefined criteria. RESULTS: Three subject indexes have been chosen--Medical Subject Headings (MeSH), CareData and the Government Category List (GCL), plus mapping between natural language and MeSH terms. This decision was a reasonable compromise between the strategy-driven aim of seamless access for all 'partners in care', and practical constraints of time/manpower. Other authority files (e.g. geographical area, language) are also standards based, and customised to reflect the information needs of an increasingly integrated health-care system. CONCLUSIONS: No single index could provide the scope required to meet the widening range of NHS information need. The influence of high-level strategic aims and objectives have extended their reach to influence indexing policy for the e-Library. Our indexing policy will continue to evolve and contribute to a knowledge management infrastructure capable of supporting current and future NHS Scotland information needs and strategy. Layperson terminology was identified as a gap; additional measures to address this gap are highlighted. PMID- 15810932 TI - SCISTER act: delivering training in information skills for social-care professionals. AB - PURPOSE: To develop and evaluate a programme of information skills training for social-care practitioners and health-care librarians. SETTING: Two one-day training courses run separately for 13 social-care practitioners and 10 health care librarians in the Trent geographic region within Northern England. METHODOLOGY: Qualitative and quantitative feedback collected through questionnaires supplemented by participant observation. RESULTS: While generally feedback was favourable, the courses were overly ambitious. More training sessions of shorter duration would offer participants opportunities to practise and use the skills they have acquired. Social-care practitioners responded more positively to skills associated with appraising qualitative research than with randomised controlled trials. Librarians appreciated the opportunity to learn about unfamiliar social-care resources. Both practitioners and health-care librarians reported that they acquired skills to support evidence-based social care as a result of the training intervention. DISCUSSION/CONCLUSION: Involvement of a multi-disciplinary team and support from a social-care information provider maximized the impact of this experimental training intervention. PMID- 15810933 TI - Evaluating the impact of Information Skills Training within primary care. PMID- 15810934 TI - Challenges to electronic journal access and funding in health sciences libraries. PMID- 15810935 TI - Sole searching--wasting our time? PMID- 15810936 TI - e-Portfolios: an introduction. PMID- 15810939 TI - Role of Helicobacter pylori in gastric carcinogenesis: the origin of gastric cancers and heterotopic proliferative glands in Mongolian gerbils. AB - Helicobacter pylori infection is well accepted to be a very important factor for the development of gastric carcinogenesis in the human stomach. In Mongolian gerbils treated with chemical carcinogens, H. pylori infection enhances glandular stomach carcinogenesis, and eradication of infection and results in curtailment of enhancing effects, particularly at early stages of associated inflammation. A high-salt diet exacerbates the effects of H. pylori infection on gastric carcinogenesis, and these two factors act synergistically to promote the development of gastric cancers in this animal model. However, the bacterium exerts the greater effects. Early acquisition significantly increases gastric chemical carcinogenesis in Mongolian gerbils, as compared to later infection. While heterotopic proliferative glands, hyperplastic and dilated glands localized beneath the muscularis mucosae, frequently develop with H. pylori infection alone in this animal model, they obviously regress on eradication, suggesting a relation to severe gastritis, rather than a malignant character. Furthermore, endocrine cells, positive for chromogranin A, are observed in the heterotopic proliferative glands, in contrast to cancerous lesions which lack endocrine elements. In conclusion, H. pylori is not an initiator, but rather a strong promoter of gastric carcinogenesis, whose eradication, together with reduction in salt intake, might effectively prevent gastric cancer development. PMID- 15810940 TI - Geographic pathology of Helicobacter pylori gastritis. AB - BACKGROUND AND AIM: Helicobacter pylori is etiologically associated with gastritis and gastric cancer. There are significant geographical differences between the clinical manifestation of H. pylori infections. The aim of this study was to compare gastric mucosal histology in relation to age among H. pylori infected patients from different geographical areas using the same grading system. The prevalence of atrophy and intestinal metaplasia were also compared with the respective gastric cancer incidence in the different countries. METHODS: A total of 1906 patients infected with H. pylori from seven countries were evaluated. Entry criteria included H. pylori positive cases with antral and corpus biopsies between the ages of 18 and 75 years. The minimum number of cases required from a country was 100. Hematoxylin-eosin stained biopsies from antrum and corpus were scored semiquantitatively using the parameters suggested by the Sydney Classification System. Statistical evaluation was performed using Kruskal Wallis test and Spearman's rank correlation test. RESULTS: The severity of gastric atrophy varied among the different groups with the highest scores being present in Japan. The lowest scores were found in four European countries and in Thailand. The scores for intestinal metaplasia were low in general except for Xi an, Japan, and Shanghai. For all the countries, the presence of atrophy in the antrum correlated well (r = 0.891) with the incidence of gastric cancer. CONCLUSION: Using a standardized grading system in a large study of H. pylori related geographic pathology, we found major differences in the overall prevalence and severity of H. pylori gastritis in relation to age. These differences mirrored the respective incidences of gastric cancer in those geographical areas. PMID- 15810941 TI - Visualization of Helicobacter species within the murine cecal mucosa using specific fluorescence in situ hybridization. AB - BACKGROUND: Members of the genus Helicobacter have been associated with colitis development in a number of immunodeficient animal models. While it is known that these organisms can initiate colitis development, the location and spatial distribution of these bacteria within the intestinal tract is currently unknown. In this study, we developed and optimized fluorescence in situ hybridization (FISH) probes specifically for Helicobacter species. MATERIALS AND METHODS: Based on 16S-RNA gene alignments, two probes specific for the entire family Helicobacteraceae and two probes specific for Helicobacter ganmani and Helicobacter hepaticus were designed. Evaluation of these probes was determined using ATCC reference strains and cecum samples from ten IL-10 knockout mice. The presence of Helicobacter species was determined using FISH and verified using PCR DGGE and microscopic examination of silver stained sections. RESULTS: Analysis of the ATCC reference strains revealed that the probes HEL274/HEL717 were specific for the family Helicobacteraceae, while HEP642 was specific for H. hepaticus and GAN1237 for H. ganmani. Using these probes, a pattern of spatial localization of the two different Helicobacter species was observed in the cecum tissues of IL-10 knockout mice. This consistently showed that H. ganmani was localized to the lower regions and H. hepaticus to the mid-upper regions of the crypts. CONCLUSION: We have developed FISH probes specific for the family Helicobacteraceae as well as two individual Helicobacter species. This study will allow the future use of the FISH to better understand host-pathogen interactions in vitro. PMID- 15810942 TI - Characteristics of a clinical isolate of urease-negative Helicobacter pylori and its ability to induce gastric ulcers in Mongolian gerbils. AB - BACKGROUND: We clinically obtained urease-negative mutant strains of Helicobacter pylori. The goal of this study was to investigate the ability of the urease negative strain to colonize and subsequently damage the gastric mucosa in Mongolian gerbils. In addition, the genes encoding the urease production in the test strain were analyzed, and other genes encoding the virulence factors, cytotoxin-associated protein and vacuolating-cytotoxin were evaluated. MATERIALS AND METHODS: The character of urease-negative isolates of H. pylori was defined. The identification of H. pylori was confirmed by polymerase chain reaction (PCR). The H. pylori isolate was transfected into Mongolian gerbils as previously described, which were followed up to 42 weeks, and the changes in their gastric mucosa were examined histologically. RESULTS AND CONCLUSION: Fifteen Mongolian gerbils orally infected with 10(7) colony forming units of urease-negative H. pylori were killed at 4, 12, 24, 36 and 42 weeks (n = 3) after infection. Culture medium without urease-negative H. pylori was given to the Mongolian gerbils as control. H. pylori continued to exist in the subject's stomach and gastric ulceration was observed and compared with the control. Clinically obtained urease negative H. pylori continued to exist for at least 42 weeks in the subject's stomach and it induced gastric ulcers. These data demonstrated that the urease in H. pylori was not a necessary factor in the formation of gastric ulcers in the Mongolian gerbil model. PMID- 15810943 TI - Acidic conditions enhance bactericidal effects of sodium bisulfite on Helicobacter pylori. AB - BACKGROUND: The Brucella broth medium, which is often used for the cultivation of microaerobic bacteria including Helicobacter pylori. It contains sodium bisulfite to decrease oxygen content in the medium. The growth of H. pylori, however, is inhibited by sodium bisulfite. In this study, the effect of sodium bisulfite was compared with several antioxidants and quantified under acidic conditions, mimicking the gastric environment. METHODS: Growth of H. pylori in the presence of several antioxidants was evaluated at OD655 nm. Effect of sodium bisulfite on H. pylori under acidic conditions was evaluated by measuring colony forming units (cfu). RESULTS: Under neutral conditions, sodium bisulfite was a more potent suppressor of H. pylori. Resveratrol, a polyphenol found in wine, exhibited the most potent inhibitory activity. To quantify the effect of sodium bisulfite on H. pylori under acidic conditions, the bacteria were grown at 37 degrees C for 30 minutes in 0.15 mol/l HCl/KCl (pH 2.0) with or without urea and sodium bisulfite. Sodium bisulfite (0.5 mmol/l) did not affect the viability at neutral pH 7.0, however, it killed H. pylori under acidic conditions, even if urea, the key substance enabling H. pylori to survive under acidic conditions, was present. The bacteria, which had been incubated under acidic conditions in the presence of urea, could survive a subsequent 30 minute-incubation at pH 2.0 without urea. Presence of sodium bisulfite, however, in the subsequent 30 minute-incubation, killed the bacteria. CONCLUSIONS: The bactericidal effect of sodium bisulfite on H. pylori was greater under acidic conditions and independent of urease activity. PMID- 15810944 TI - Effect of the carbonic anhydrase inhibitor, acetazolamide, on Helicobacter pylori infection in vivo: a pilot study. AB - BACKGROUND: Carbonic anhydrase inhibitors have been successfully used to treat peptic ulcers. Although carbonic anhydrase restriction does not inhibit Helicobacter pylori in vitro, recent studies suggest that carbonic anhydrase inhibition reduces the ability of H. pylori to survive in an acid environment as present in the stomach. METHODS: In a pilot study, we examined the effect of acetazolamide 500 mg as twice a day for 4 days in volunteers with active H. pylori infection. Effectiveness was judged by changes in the results of the urea breath test. RESULTS: Eight H. pylori infected volunteers completed the test. No urea breath test reverted to negative and there was a trend for the urea breath test value to increase [e.g. delta over baseline (DOB) mean +/- SE increased from 50.9 +/- 13 at baseline to 64.9 +/- 13 at day 5] during treatment with acetazolamide. CONCLUSION: The potential effect of carbonic anhydrase inhibitors on acid secretion may prevent effect on H. pylori in vivo and/or the sites of infection at the surface of the stomach may have a pH higher for any postulated acid-dependent effect to have an effect clinically. PMID- 15810945 TI - Efficacy of cranberry juice on Helicobacter pylori infection: a double-blind, randomized placebo-controlled trial. AB - BACKGROUND: Helicobacter pylori infection is a major cause of peptic ulcer disease and gastric cancer. This study postulated that cranberry juice would be effective in the suppression of H. pylori in an endemically infected population at high risk for gastric cancer. MATERIALS AND METHODS: A prospective, randomized, double-blind, placebo-controlled trial was conducted in Linqu County of Shandong Province, China, where 189 adults aged 48.9 +/- 11.2 years (mean +/- SD) with H. pylori infection were randomly divided into two groups: cranberry juice (n = 97) and placebo (n = 92). Participants were assigned to orally receive two 250-ml juice boxes of cranberry juice or matching placebo beverage daily for 90 days. The degree of H. pylori infection was determined using the 13C-urea breath test before randomization at 35 and 90 days of intervention to assess the efficacy of cranberry juice in alleviating infection. RESULTS: A total of 189 subjects with positive 13C-urea breath test results prior to randomization completed the study. At day 35 of intervention, 14 of the 97 (14.43%) from the the cranberry juice treatment group and 5 of the 92 (5.44%) of the placebo recipients had negative 13C-urea breath test results. After 90 days, the study concluded that 14 of the 97 subjects in the cranberry juice treatment group versus 5 of the 92 in the placebo group yielded negative test results. Eleven individuals from the cranberry juice treatment group and only two from the placebo group were negative at 35 and 90 days of experiment. These results are significant (p < .05). CONCLUSIONS: Regular consumption of cranberry juice can suppress H. pylori infection in endemically afflicted populations. PMID- 15810946 TI - Lewis epitopes on outer membrane vesicles of relevance to Helicobacter pylori pathogenesis. AB - BACKGROUND: Helicobacter pylori extrudes protein- and lipopolysaccharide-enriched outer membrane vesicles from its cell surface which have been postulated to act to deliver virulence factors to the host. Lewis antigen expression by lipopolysaccharide of H. pylori cells has been implicated in a number of pathogenic roles. The aim of this study was to further characterize the expression of lipopolysaccharide on the surface of these outer membrane vesicles and, in particular, expression of Lewis antigens and their association with antibody production in the host. MATERIALS AND METHODS: H. pylori strains were examined for outer membrane vesicle production using transmission electron microscopy and Lewis antigen expression probed using immunoelectron microscopy. Sera from patients were analyzed for cross-reacting anti-Lewis antibodies and, subsequently, absorbed using outer membrane vesicle preparations to remove the cross-reacting antibodies. RESULTS: The formation of outer membrane vesicles by H. pylori was observed in both in vitro and in vivo samples. Furthermore, vesicles were produced following culture in either liquid or solid medium by all strains examined. Moreover, we observed the presence of Lewis epitopes on outer membrane vesicles using immunoelectron microscopy and immunoblotting. Circulating anti-Lewis antibodies were found in the sera of gastric cancer patients but not in the sera of H. pylori-negative control subjects. Absorption of patient sera with outer membrane vesicles decreased the levels of anti-Lewis autoantibodies. CONCLUSIONS: Our results demonstrate the ability of H. pylori to generate outer membrane vesicles bearing serologically recognizable Lewis antigens on lipopolysaccharide molecules which may contribute to the chronic immune stimulation of the host. The ability of these vesicles to absorb anti-Lewis autoantibodies indicates that they may, in part, play a role in putative autoimmune aspects of H. pylori pathogenesis. PMID- 15810947 TI - Controversies of the cardiac mucosa and Barrett's oesophagus. AB - Confusion regarding the diagnosis of Barrett's oesophagus exists because of a false dogma that cardiac mucosa is normally present in the gastro-oesophageal junctional region. Recent data indicate that the only normal epithelia in the oesophagus and proximal stomach are squamous epithelium and gastric oxyntic mucosa. When this fact is recognized, it becomes easy to develop precise histological definitions for the normal state (presence of only squamous and oxyntic mucosa), metaplastic oesophageal columnar epithelium (cardiac mucosa with and without intestinal metaplasia, and oxynto-cardiac mucosa), the gastro oesophageal junction (the proximal limit of gastric oxyntic mucosa), the oesophagus (that part of the foregut lined by squamous and metaplastic columnar epithelium), reflux disease (the presence of metaplastic columnar epithelium), and Barrett's oesophagus (cardiac mucosa with intestinal metaplasia). It is also possible to assess accurately the severity of reflux which is directly proportional to the amount of metaplastic columnar epithelium, and the risk of adenocarcinoma which is related to the amount of dysplasia in intestinal metaplastic epithelium present within the columnar lined segment of the oesophagus. Histopathological precision cannot be matched by any other modality and can convert the confusion that exists regarding diagnosis of Barrett's oesophagus to complete lucidity in a manner that is simple, accurate, and reproducible. PMID- 15810948 TI - Severe gastritis in the Peruvian Andes. AB - OBJECTIVE: To compare the Helicobacter pylori-associated pathology in gastric biopsies taken from patients living at sea level with those taken from patients living at high altitude. METHODS AND RESULTS: We included 38 patients from a hospital in the Andean city of La Oroya, Peru, located at 3700 m in altitude, and 40 control patients taken from Comas Clinic located in the city of Lima at sea level. Fibrepanendoscopy and multiple biopsies were performed in all the patients followed by histopathological examination. In the antrum, patients from the Andean town had a higher prevalence of glandular lymphoid adherence lesions, active germinal centres, moderate to severe chronic atrophic gastritis, intestinal metaplasia and moderate to severe total deep gland loss, than did patients from the coastal town. Furthermore, the severity of the histological lesions seen in the gastric body and cardia was significantly greater in the high altitude patients than in those from sea level. CONCLUSION: This study suggests that the severity of H. pylori-associated gastric lesions seen on histopathological examination is greater in patients living at high altitude, the cause of which is most probably multifactorial but nonetheless principally altitude related. PMID- 15810949 TI - High tissue expression of tumour-associated trypsin inhibitor (TATI) associates with a more favourable prognosis in gastric cancer. AB - AIMS: The tumour-associated trypsin inhibitor (TATI) is a 6-kDa protease inhibitor with potential inhibitory effects on tissue degradation. In serum, increased levels have been associated with adverse prognosis in different forms of cancer. We assessed the tumour tissue expression and prognostic value of TATI in a surgically treated, single-institution series of patients with gastric cancer. METHODS AND RESULTS: Using a monoclonal anti-TATI antibody, immunohistochemistry was performed on formalin-fixed paraffin-embedded tumour specimens from 336 patients. TATI expression was observed in 265 (79%) of the tumours. There was a significant association between high TATI expression and low stage (P = 0.007), superficial tumours (P = 0.005), and absence of nodal (P = 0.015) and of distant metastases (P = 0.022). In univariate analysis, patients with high TATI expression had a significantly more favourable 5-year cumulative survival compared with patients with negative to moderate immunostaining (43% and 28%, respectively, P = 0.006). On multivariate survival analysis stratified for estimated cure of surgery, stage (P < 0.0001) and age (P = 0.022) at the time of surgery were independent prognostic factors. CONCLUSIONS: High TATI expression in tumour tissue was detected more frequently in patients with early-stage gastric cancer and seems to correlate with a favourable outcome. PMID- 15810950 TI - Multidrug resistance-associated protein (MRP1) gene is strongly expressed in gastric carcinomas. Analysis by immunohistochemistry and real-time quantitative RT-PCR. AB - AIMS: To assess MRP1 protein and MRP1 mRNA levels in gastric carcinomas and in non-neoplastic mucosa remote from the tumours. MRP1 gene expression may play a role in the complex pattern of chemoresistance present in gastric carcinomas. METHODS AND RESULTS: A total of 57 carcinomas and respective gastric tissues were included for immunohistochemical assessment with the anti-MRP1 monoclonal antibodies MRPr1 and QCRL-1. Of these, 35 tumour and gastric mucosa tissues were also assessed by real-time quantitative reverse transcriptase-polymerase chain reaction. Medium or high MRP1 protein expression was detected in 89% and 77% of carcinomas and in 96% and 93% of non-neoplastic gastric mucosa by MRPr1and QCRL 1, respectively. No difference in MRP1 mRNA levels was detected between carcinomas and non-neoplastic gastric mucosa tissues in 77% of the patients. A significant correlation was found between MRP1 mRNA level and protein expression detected in carcinomas related to non-neoplastic gastric mucosa, although they were non-concordant in 29% of the patients. CONCLUSIONS: MRP1 gene is usually expressed in most gastric carcinomas and does not differ substantially from that observed in non-neoplastic gastric mucosa remote from the tumour. However, a decrease in MRP1 gene expression is found in some carcinomas. For accurate assessment of changes in MRP1 expression between tumour and normal tissues both protein and mRNA detection are necessary. PMID- 15810951 TI - Expression of D2-40 in lymphatic endothelium of normal tissues and in vascular tumours. AB - AIMS: To evaluate the expression of D2-40 in normal lymphatic endothelium and vascular tumours or tumour-like lesions of the skin and soft tissue. D2-40 is a novel monoclonal antibody to a Mr 40 000 O-linked sialoglycoprotein that reacts with a fixation-resistant epitope in lymphatic endothelium. METHODS AND RESULTS: Formalin-fixed paraffin-embedded sections from 30 normal tissue samples, including skin, soft tissue, stomach, and colon, and 84 vascular tumours or vascular tumour-like lesions were immunostained with monoclonal antibodies to D2 40 and CD31. Normal lymphatic endothelial cells in all normal tissues expressed D2-40. Its positive staining delineated flattened channels or open spaces lined by a single layer of endothelial cells whose lumena were sometimes filled with lymphocytes. Ten of 10 cases of lymphangioma, nine of 10 Kaposi's sarcomas (KSs), one of five spindle cell haemangiomas, one of one reactive angioenodotheliomatosis, one of one vascular transformation of lymph node sinuses, three of three Dabska tumours, one of 10 epithelioid haemangioendotheliomas (HEs) and seven of 15 angiosarcomas were positive for D2 40. Positively staining angiosarcomas were characterized by epithelioid or papillary endothelial cells. Twenty-two non-spindle cell haemangiomas, one retiform HE and one Kaposiform HE, and five glomus tumours were negative for D2 40. In comparison, CD31 was expressed in five of 10 lymphangiomas, nine of 10 KSs, 27 of 27 haemangiomas, three of three Dabska tumours, 10 of 10 epithelioid HEs, 15 of 15 angiosarcomas and one of one each of retiform HE, Kaposiform HE, reactive angioendotheliomatosis, and vascular transformation of node sinuses. Five glomus tumours were negative for CD31. CONCLUSIONS: The monoclonal antibody D2-40 is a highly sensitive and specific marker of lymphatic endothelium in normal tissue and a subset of vascular lesions, including KS, Dabska tumour, and lymphangioma. The findings support the concept that these tumours show at least partial lymphatic endothelial differentiation. Subsets of angiosarcomas and HEs show both vascular and lymphatic endothelial differentiation. D2-40 can be used in a panel of markers to classify vascular tumours. There is no requirement for epitope retrieval. This novel monoclonal antibody also has the potential for increasing the accuracy of detection of lymphatic invasion in primary tumours and could be widely applied for this purpose in surgical pathology. PMID- 15810952 TI - Supratentorial primitive neuroectodermal tumours of the brain: multidirectional differentiation does not influence prognosis. A clinicopathological report of 18 patients. AB - AIMS: To examine the clinical and pathological characteristics of supratentorial primitive neuroectodermal tumours (PNETs) in a retrospective series of 18 patients, according to the strict definition of the World Health Organization classification of tumours that excludes other types of malignant embryonal tumours of the brain. METHODS AND RESULTS: Eleven children and seven adults with supratentorial PNETs were diagnosed between 1993 and 2002 and their medical records were reviewed. An immunohistochemical study was performed on formalin fixed paraffin-embedded tissue of 18 primary tumours and five recurrences with antibodies for neuronal (neuron specific enolase, synaptophysin, neurofilament, chromogranin A), epithelial [epithelial membrane antigen (EMA), cytokeratin], glial [glial fibrillary acidic protein (GFAP)], muscle (desmin, h-caldesmon, alpha-smooth muscle actin, myogenin) differentiation and with two anti-CD99 antibodies. All tumours showed at least one neuronal marker except chromogranin A; a variable number of cells were GFAP+ or EMA+ in 18/23 tumours. Six primary tumours and one recurrence were positive for cytokeratin and/or one muscle antigen except myogenin. CD99 was observed in 33% of the cases. The mean duration of overall survival was 20 months. The estimated overall survival rates were 61% at 1 year, 29% at 2 years, and 18% at 3 years. Two factors of poor prognosis were identified by univariate analysis: a positive cerebrospinal fluid cytology at diagnosis and the absence of complete resection. No distinct immunophenotype was statistically related to survival. CONCLUSIONS: A multidirectional differentiation is a frequent event in supratentorial PNETs but has no apparent influence on the outcome of this aggressive neoplasm. PMID- 15810953 TI - Collagen and elastic system in the remodelling process of major types of idiopathic interstitial pneumonias (IIP). AB - AIMS: Structural remodelling in acute and chronic idiopathic interstitial pneumonia (IIP) has been extensively investigated, but little attention has been directed to the elastic tissue in these situations. The aim of this study was to determine whether elastic deposition accompanies collagen deposition in the four major histological patterns of IIP: diffuse alveolar damage (DAD), organizing pneumonia (OP), non-specific interstitial pneumonia (NSIP) and usual interstitial pneumonia (UIP). METHODS AND RESULTS: We measured, by image analysis, the content of fibres of the collagenous and elastic systems of the alveolar septum in histological slides of open lung biopsies, using the picrosirius-polarization method and Weigert's resorcin-fuchsin stain, respectively. Five groups were studied: 10 cases of DAD; nine cases of OP; nine cases of NSIP; and 10 cases of UIP. Four normal lungs were used for comparison. The content of collagen fibres was significantly higher in UIP when compared to DAD, NSIP, OP and normal lung. The content of elastic fibres was increased in comparison with normal lungs but this was not significantly different among the histological patterns. CONCLUSION: Acute and chronic IIP cause a similar increase in the collagen and elastic contents of the lungs, representing a process of 'fibroelastosis' rather than an exclusive process of fibrosis. A profibrogenic mechanism is responsible for the unparallelled collagen augmentation observed in UIP subjects, the nature of which is yet to be determined. PMID- 15810954 TI - Assessing microvessels and angiogenesis in human breast cancer, using VE cadherin. AB - AIMS: Vascular endothelial (VE)-cadherin, also known as cadherin-5 and CD144, is an adhesion molecule uniquely expressed in endothelial cells. We hypothesized that VE-cadherin may be a useful marker for assessing microvessels and angiogenesis in human breast cancer and sought to determine whether a correlation exists between levels of VE-cadherin, angiogenic markers factor VIII and platelet endothelial cell adhesion molecule (PECAM)-1 and patient outcome in breast cancer. METHODS AND RESULTS: Frozen sections from breast cancer primary tumours (tumour n = 114, background n = 30) were immunostained with VE-cadherin, factor VIII and PECAM-1 antibodies and microvessel number was assessed. RNA was reverse transcribed and analysed by quantitative polymerase chain reaction (Q-PCR). VE cadherin immunostaining showed a significant difference in microvessel number in tumour compared with background. There was no significant difference in the number of microvessels stained with PECAM-1 or factor VIII; there was increased staining of other structures within the sample and higher general background staining. Q-PCR revealed elevated levels of VE-cadherin and PECAM-1 in tumour samples compared with background tissue and in patients with a poor prognosis, as determined by the Nottingham Prognostic Index. There was no difference in levels with factor VIII. Both VE-cadherin and PECAM-1 had significantly reduced expression in lobular compared with ductal carcinomas: there was no difference with factor VIII. CONCLUSION: Higher levels of angiogenic marker molecules in breast cancer may have an association with poor prognosis in patients. Moreover, VE-cadherin appears to be a preferable marker for such analysis. PMID- 15810955 TI - Real-time reverse transcription-PCR and fluorescence in-situ hybridization are complementary to understand the mechanisms involved in HER-2/neu overexpression in human breast carcinomas. AB - AIMS: To evaluate the HER-2/neu status at the mRNA and DNA level of breast carcinomas and to compare it with HER-2/neu receptor overexpression by immunohistochemistry (IHC). METHODS AND RESULTS: In 32 invasive breast carcinomas, frozen tissue was available for real-time detection of HER-2/neu mRNA levels by reverse transcription-polymerase chain reaction (RT-PCR). Corresponding paraffin sections were examined by IHC and fluorescence in-situ hybridization (FISH). Thereby, different IHC and FISH procedures were compared. Using microwave epitope retrieval, all 32 cases scored 3+ on IHC, whereas only 28 out of 32 cases scored IHC 3+ using water bath epitope retrieval. All of these 28 cases showed increased levels of HER-2/neu mRNA. Dual-colour FISH analysis showed corresponding gene amplification in all 28 cases, with two cases showing a peculiar amplification pattern. In the remaining four cases, scoring IHC 2+ using water bath epitope retrieval, mRNA levels were not elevated. Three cases did not have gene amplification and one case showed low-level HER-2/neu gene amplification. All four carcinomas showed chromosome 17 polysomy. CONCLUSIONS: Real-time RT-PCR is accurate in selecting breast carcinoma cases scoring 3+ by IHC with high-level gene amplification. Results obtained by dual-colour FISH suggest that mechanisms leading to HER-2/neu receptor overexpression may be different between carcinomas scoring 2+ and 3+ on IHC, with polysomy 17 found in the former and gene amplification in the latter. PMID- 15810956 TI - Terminal deoxynucleotidyl transferase-positive cells in trephine biopsies following bone marrow or peripheral stem cell transplantation reflect vigorous B cell generation. AB - AIMS: Bone marrow is the major site of B-cell generation in humans. While in early childhood a high number of B-cell precursors is found in the bone marrow, only very few such cells are usually detectable in adult bone marrow. To assess the number of immature B cells present after haematopoietic cell transplantation the number of terminal deoxynucleotidyl transferase (TdT)-positive cells in regenerating bone marrow of adult patients was analysed. METHODS AND RESULTS: Bone marrow biopsy specimens were analysed from patients after allogeneic bone marrow transplantation (BMT; n = 14) or stem cell transplantation (SCT; n = 25) and autologous BMT (n = 9). Specimens from 11 untransplanted adult patients and 11 infants were also studied, as negative and positive controls, respectively. Immunohistochemistry was performed on paraffin-embedded bone marrow biopsy sections using TdT as a marker of lymphoid progenitors. Immunoreactivity for CD79a, CD20 and CD10 was used to confirm their B-cell origin. Using computer assisted automated image analysis we quantitatively assessed the TdT+ cells present. We found a significant increase in the numbers of B-cell precursors in the bone marrow after allogeneic and autologous BMT/SCT compared with adult controls (P = 0.022). To analyse this in detail, we followed some patients after allogeneic BMT/SCT for up to 1445 days, when a marked B-cell increase was still detectable. However, the median number of TdT+ B cells after BMT/SCT was significantly lower than the number of equivalent B cells in infantile bone marrow biopsy specimens (P < 0.001). CONCLUSIONS: Bone marrow of adult patients after BMT/SCT is capable of initiating vigorous precursor B-cell generation, which is not seen in untransplanted adults. However, the increase of immature B cells was variable in our study. Only in two young adult patients did it reach the magnitude of B-cell generation seen in infantile bone marrow where immunocompetent B cells are produced normally. A marked increase in number of immature B cells post-transplant may mimic B-cell acute lymphoblastic leukaemia (B-ALL). This is a potential problem in patients transplanted for B-ALL itself. Since reactive and neoplastic B-cell precursors share the same immunophenotype in paraffin-embedded tissue, additional tools, particularly molecular techniques, may have to be employed to establish the correct diagnosis. PMID- 15810957 TI - Morphometric analysis of primary graft non-function in liver transplantation. AB - AIMS: Primary graft non-function (PNF) is a life-threatening condition that is thought to be the consequence of microcirculation injury. The aim of the present study was to assess, with a computerized morphometric model, the morphological changes at reperfusion in liver biopsy specimens from patients who developed PNF after liver transplantation. METHODS AND RESULTS: Biopsy specimens were obtained at maximum ischaemia and at the end of reperfusion. Morphology included many stereological parameters, such as volumes of all parenchymal components, surface density, size distribution and mean diameter of hepatocytes. Other variables examined were intensive care unit stay, degree of steatosis, serum liver function tests and ischaemic time. In the postoperative period, the PNF group showed elevated serum levels of alanine transferase, decreased daily rate of bile production and prothrombin activity. Blood lactates were significantly higher in the PNF group than in a control group. When comparing groups, the volumetric parameters related to hepatocytes and sinusoids and the surface densities of the hepatic cells showed an inverse relationship. At the end of reperfusion, in PNF group the volume fraction of hepatocyte cytoplasm was decreased; in contrast, the volume fraction of sinusoidal lumen was markedly increased. The cell profiles showed the same inverse trend: the surface density of the parenchymal border of hepatocytes was decreased in PNF when compared with the control group, while the surface density of the vascular border was increased. In the PNF group, the surface density of the sinusoidal bed was directly correlated with alanine transferase, daily rate of bile production, prothrombin activity and cold ischaemic time. CONCLUSIONS: The alterations in hepatic architecture, as demonstrated by morphometric analysis in liver transplant recipients that developed PNF, provide additional information that may represent useful viability markers of the graft to complement conventional histological analysis. PMID- 15810958 TI - Isolated intramucosal goblet cells in subacute ischaemic enteritis: mimicry of signet ring cell carcinoma. PMID- 15810960 TI - Angioimmunoblastic T-cell lymphoma with hyperplastic germinal centres and a high content of EBV-infected large B-cells carrying IgH chain gene monoclonal rearrangement. PMID- 15810961 TI - Detection of t(14;18) translocation in a case of intravascular large B-cell lymphoma: a germinal centre cell origin in a subset of these lymphomas? PMID- 15810962 TI - WT-1 in ovarian and endometrioid serous carcinoma: a meta-analysis. PMID- 15810963 TI - Metastasizing myxopapillary ependymoma. PMID- 15810964 TI - Pathological changes related to Imatinib treatment in a patient with a metastatic gastrointestinal stromal tumour. PMID- 15810965 TI - An unusual case of clear cell sarcoma arising in the jejunum highlights the diagnostic value of molecular genetic techniques in establishing a correct diagnosis. PMID- 15810966 TI - Malacoplakia of the gallbladder. PMID- 15810967 TI - High grade prostatic intraepithelial neoplasia involving small ducts and acini. PMID- 15810969 TI - Behaviour of bone marrow osteoblast-like cells on mineral trioxide aggregate: morphology and expression of type I collagen and bone-related protein mRNAs. AB - AIM: To investigate the in vitro behaviour of rat bone marrow cells (RBM) on mineral trioxide aggregate (MTA) (ProRoot, MTA Root Canal Repair Material; Dentsply Tulsa, Tulsa, OK, USA) compared with intermediate restorative materials (IRM) (Dentsply Caulk, Milford, DE, USA). METHODOLOGY: RBM were obtained from rat femur and were primary cultured and then subcultured. Cells were then seeded on three dishes of each material, and cultured for 3 days, after which they were evaluated morphologically using scanning (SEM) and transmission (TEM) electron microscopy. Furthermore, the calcium released from hydrated material, the cell proliferation ratio and alkaline phosphatase (ALP) activity were analysed, and the expression of type I collagen and bone-related protein mRNAs were evaluated. The data were averaged and analysed via one-way analysis of variance (anova) and were then compared by the Scheffe's test. RESULTS: SEM showed that RBM attached to MTA and had a flattened appearance without nuclear protrusions and microspikes. TEM showed that the cells attached in the same manner as the control group, but gaps larger than 2 microm were frequently seen. The calcium released from hydrated MTA was about 130 ppm after 3 days of immersion in saline. The ALP activity was similar to the control group. Cell proliferation and expression of type I collagen mRNA was significantly lower, while the expression of osteopontin mRNA was significantly higher than the control group at the third day of culture. In IRM groups, a few rounded cells were observed on the material but no living cells were seen. CONCLUSIONS: MTA is a material of low toxicity which does not inhibit cell growth, but does suppress the differentiation of osteoblast-like cells. PMID- 15810970 TI - Effect of bleaching agents on bonding to pulp chamber dentine. AB - AIM: To determine the effect of intracoronal bleaching agents on adhesion of bonding agents to pulp chamber dentine. METHODOLOGY: Forty extracted human maxillary anterior teeth were randomly divided into four groups of 10 teeth each. Bleaching agents were sealed in pulp chambers for 7 days, as in clinical use. Group 1 (control): distilled water, group 2: 35% hydrogen peroxide, group 3: sodium perborate mixed with water, and group 4: sodium perborate mixed with 35% hydrogen peroxide. Teeth were stored in saline at 37 degrees C for 7 days. After the bleaching agent was removed, teeth were leached in water for a further 7 days prior to bonding. The crown was cut vertically from mesial to distal and the labial pulp chamber dentine was prepared for bonding with Clearfil SE-Bond and filled with resin composite (Clearfil AP-X). The bonded specimens were kept moist at 37 degrees C for 24 h. Microtensile bond strengths were determined using a universal testing machine. Additional teeth were prepared using the same bleaching procedures to investigate the scanning electron microscopic appearance of the dentine surface. RESULTS: Mean values (+/-SD) of microtensile bond strength for the experimental groups were: group 1: 5.29 +/- 2.21 MPa, group 2: 5.99 +/- 1.51 MPa, group 3: 9.17 +/- 1.65 MPa and group 4: 3.99 +/- 1.31 MPa. Dentine treated with sodium perborate in water (group 3) had significantly higher mean bond strength when compared with the other three groups (P < 0.05, Tukey's test). Mean bond strength was lowest when dentine was treated with sodium perborate plus hydrogen peroxide (group 4). CONCLUSIONS: In terms of subsequent bond strength during restoration, sodium perborate mixed with distilled water appears to be the best intracoronal bleaching agent. PMID- 15810971 TI - Intracanal temperature rise evaluation during the usage of the System B: replication of intracanal anatomy. AB - AIM: To evaluate and determine intracanal temperature rises at 2 and 4 mm from the working length (WL) necessary to obtain proper replication of intracanal anatomy with gutta-percha (GP) using the System B heat source during vertical condensation. METHODOLOGY: A split-tooth model was prepared and artificial shallow depressions were cut in the buccal canal wall 2 and 4 mm from the WL. At the same level on the palatal wall holes were drilled to adapt two thermocouples. The canal was filled using GP in a vertical condensation technique by placing the System B plugger at 2 and 4 mm from the WL in groups A and B, respectively. Two control groups in which no GP was used were carried out placing the plugger 2 and 4 mm from the WL (groups A.c and B.c, respectively) and activating the heat source. Recording of temperature rise was carried out during the filling procedure for groups A and B and during activation for control groups A.c and B.c; the highest temperatures were recorded. After each filling was completed, 3 min were allowed for the GP to cool and the model divided to reveal the filling. Images of the GP were taken with a CCD camera to evaluate the presence of replication of artificial round depressions. In control groups, the temperature was recorded for 20 s after a 3 s activation of the heat source. The rise in temperature was compared between the groups individually at each level (2 or 4 mm) and statistically analysed using one-way anova and Fisher PLSD tests at 5% of significance level (P < 0.05). RESULTS: Mean temperature rises of 14 +/- 3 and 12 +/- 2 degrees C at 2 and 4 mm from the WL, respectively, were observed in group A fillings, and 4 +/- 1 and 6 +/- 1 degrees C at 2 and 4 mm, respectively, in group B fillings. Recordings at 2 mm showed significantly (P < 0.05) higher temperature rises with group A.c when compared with groups B and B.c. Replication of intracanal anatomy with GP was always found in group A fillings at both levels but only 4 mm from the WL in group B fillings. CONCLUSIONS: Positioning the plugger close to WL and a temperature rise of 6 degrees C were necessary to obtain replication of intracanal anatomy. A mean temperature rise of 4 degrees C at 2 mm from WL (group B) resulted in no replication of intracanal anatomy. Further studies simulating clinical conditions are necessary. PMID- 15810972 TI - Effect of liquid and paste-type lubricants on torque values during simulated rotary root canal instrumentation. AB - AIM: To evaluate the effects of lubrication on torque generated during rotary preparation of simulated root canals in dentine using ProFile and ProTaper Nickel Titanium instruments. METHODOLOGY: Thirty-two 3 mm thick dentine discs were used for this study. Four pilot holes were drilled perpendicularly through each disc. These simulated root canals were filled with water, aqueous 15% ethylenediamine tetraacetic acid (EDTA) or a paste-type lubricant (Glyde). Empty canals served as controls. Peak torque values and apically directed forces of rotary instruments were measured using a specially designed testing platform. Full torsional loads over time were calculated by integration of continuous torque records. Data were compared using analyses of variance with the alpha-type error set at 0.05. RESULTS: Maximum torque values using ProFile instruments were significantly (P < 0.005) reduced by the EDTA solution, whilst no reduction was observed using Glyde. With ProTaper instruments, no lubricant effects on maximum torque values were observed (P = 0.372). Full torsional loads over time were significantly reduced for both ProFile and ProTaper preparation (P < 0.005) by the use of lubrication. The two fluids performed significantly better than paste-based Glyde. All lubricants reduced force when applied to ProTaper by mean reductions ranging from 12 to 54%, whilst Glyde lead to increased force when applied to ProFile. CONCLUSIONS: Under the conditions of this laboratory study, lubrication appeared to be advantageous. Aqueous solutions generally performed better than the paste-type product under investigation, which showed untoward effects when used in conjunction with a rotary instrument with a U-shaped cross-section (ProFile). PMID- 15810973 TI - Ex vivo fracture resistance of direct resin composite complete crowns with and without posts on maxillary premolars. AB - AIM: To investigate ex vivo the fracture resistance and failure mode of direct resin composite complete crowns with and without various root canal posts made on maxillary premolars. METHODOLOGY: The clinical crowns of 40 human extracted single-rooted maxillary premolars were sectioned at the cemento-enamel junction. The canals were prepared with Gates Glidden drills up to size 4. Thirty samples were provided with standardized post spaces in the palatal canal and all roots were embedded in acrylic. Minimal standardized preparations in the canal entrances were made. Groups of 10 samples were treated with (i) prefabricated metal posts, (ii) prefabricated glass fibre posts, (iii) custom-made glass fibre posts, and (iv) no posts (control). Posts were cemented with resin cement and resin composite complete crowns were made. All specimens were thermocycled (6000x, 5-55 degrees C). Static load until fracture was applied using a universal loading device (crosshead speed 5 mm min(-1)) at a loading angle of 30 degrees . Failure modes were categorized as favourable and unfavourable failures. RESULTS: No significant difference was observed between the mean failure loads (group 1: 1386 N, group 2: 1276 N, group 3: 1281 N, and group 4: 1717 N, P > 0.05), nor between frequencies of failure modes (P > 0.05). All failures were fractures of the resin composite crown in combination with tooth material (cohesive failures). CONCLUSIONS: Within the limits of this laboratory investigation it is concluded that severely damaged and root filled maxillary premolars, restored with direct resin composite complete crowns without posts have similar fracture resistances and failure modes compared to those with various posts, which suggest that posts are not necessarily required. PMID- 15810974 TI - Prevalence of apical periodontitis and the quality of endodontic treatment in an adult Belarusian population. AB - AIM: To estimate the prevalence of teeth with apical periodontitis (AP) and technically failed root fillings in an adult Belarusian population. METHODOLOGY: Panoramic radiographs of all 1423 patients over 15 years of age not seeking emergency dental care, and attending the Dental School of the Belarusian Medical University for the first time during the period from 1 January to 31 December 2001 were examined. The quality of root fillings was scored according to criteria of length proposed by De Moor et al. [International Endodontic Journal 33 (2000) 113] and the periapical status of all teeth (except third molars) was categorized on the basis of presence or absence of radiographic signs of AP. The data were analysed using the chi-square test and odds ratio. RESULTS: Radiographs indicated that 8632 teeth (22% in the maxilla; 21% in the mandible) were missing leaving a total of 31,212 teeth to be assessed. Twenty per cent of the teeth had some filling material in the root canal(s). AP was found in 1141 subjects (80%) and 12% of the teeth. AP was more frequently associated with molar teeth (23%) than premolar (14%), canine (4%) and incisor teeth (6%). AP was diagnosed in 45% of root filled teeth, the remaining cases with AP had not been root filled. Statistical analysis showed that the probability of radiological detection of AP in root filled teeth was 25-fold higher than when the root canals had not been filled (chi2 = 8636.04, P < or = 0.001, odds ratio with 95% confidence intervals: 23.01 < 25.17 < 27.45). Periapical radiolucencies with adequately filled root canals occurred significantly less often than with teeth in which the root canal was filled more than 2 mm from radiographic apex or when filling material was extruded through the apex. CONCLUSION: The prevalence of AP in all age groups in Belarus was higher than in other populations. The probability of AP increased significantly after root canal treatment and was closely correlated with the quality of the root filling. PMID- 15810975 TI - A 24-year follow-up of root filled teeth and periapical health amongst middle aged and elderly women in Goteborg, Sweden. AB - AIM: To describe the endodontic status amongst middle-aged and elderly women longitudinally and cross-sectionally over 24 years. METHODOLOGY: A random sample of 1462 women 38, 46, 50, 54 and 60 years old, living in Goteborg, Sweden, were sampled in 1968 for medical and dental examinations with a participation rate of 90.1%. The same women were re-examined in 1980 and 1992 together with new 38- and 50-year-old women. The dental examination consisted of questionnaires, clinical and panoramic radiological survey (OPG). The number of teeth, number of root filled teeth (RF) and number of teeth with periapical radiolucencies (PA) were registered. The RF and PA ratios were calculated. Cross-sectional data were analysed by means of anova and longitudinal data by a general linear model for repeated measures. Sample prevalences were compared and statistical inferences were made with the chi-squared test. In all analysis, the confidence interval (CI) regarded mean difference between groups (95% CI). RESULTS: The RF and PA ratio decreased over time as well as the frequency of edentulous subjects. Cross sectional analysis revealed a minor increase in frequency of RF and PA and loss of teeth with age. Longitudinally, loss of teeth was evident in all cohorts. In addition, there was a trend of lower number of teeth with PA, and the RF ratio increased with age. CONCLUSIONS: The prevalence of periapical disease did not increase with age, probably as a result of root canal treatment and extractions. Data showed that the prevalence of RF teeth and teeth with PA decreased for comparable age cohorts during the 24-year follow-up. PMID- 15810976 TI - Garre's osteomyelitis managed by root canal treatment of a mandibular second molar: incorporation of computed tomography with 3D reconstruction in the diagnosis and monitoring of the disease. AB - AIM: To report the healing of Garre's osteomyelitis involving a mandibular second molar following root canal treatment. SUMMARY: To describe the endodontic management of Garre's osteomyelitis involving a mandibular second molar. In this case, computed tomography (CT) was used to evaluate the status of the periapical lesion. Five years after root canal treatment, the tooth and supporting tissues appeared healthy both clinically and radiographically and were functioning well. KEY LEARNING POINTS: Garre's osteomyelitis of an adult can be managed by root canal treatment. Computed tomography could be used for diagnosis and treatment planning in endodontics. PMID- 15810977 TI - Calculus-like deposit on the apical external root surface of teeth with post treatment apical periodontitis: report of two cases. AB - AIM: To report two cases in which calculus-like material was found on external root surfaces of (i) an extracted root and (ii) an apicected part of a root, both of which were removed due to post-treatment refractory apical periodontitis. SUMMARY: In each case, there was a fistulous tract, which did not heal after conventional root canal treatment. The first case did not heal even after apical surgery, and subsequent tooth extraction revealed calculus-like material on a root surface of complex anatomy. The second case showed radiographic signs of healing after apicectomy. Histology of the apical biopsy revealed a calculus-like material on the external surface of the root apex. It is suggested that the presence of calculus on the root surfaces of teeth with periapical lesions may contribute towards the aetiology of failure. KEY LEARNING POINTS: Biofilm on the external root surface has been implicated in the failure of apical periodontitis to heal, despite adequate root canal treatment. Calculus-like material was found, in two cases, on the root surface of teeth with post-treatment apical periodontitis, where the only communication externally was a sinus tract. PMID- 15810978 TI - Embryonic reversions and lineage infidelities in tumour cells: genome-based models and role of genetic instability. AB - Reversions to "embryonic precursor"-type cells and infidelities of tumour cell lineage (including metaplasias) have been recognized as aspects of various tumour types since the 19th century. Since then, evidence of these phenomena has been obtained from numerous clinical, biochemical, immunological and molecular biological studies. In particular, microarray studies have suggested that "aberrant" expressions of relevant genes are common. An unexplained aspect of the results of these studies is that, in many tumour types, the embryonic reversion or lineage infidelity only occurs in a proportion of cases. As a parallel development during the molecular biological investigation of tumours over the last several decades, genetic instability has been found much more marked, at least in some preparations of tumour cells, than that identified by means of previous karyotypic investigations of tumours. This study reviews examples of embryonic reversion and lineage infidelity phenomena, which have derived from the various lines of investigation of cancer over the last 150 or so years. Four categories of circumstances of the occurrence of embryonic reversions or lineage infidelities have been identified - (i) as part of the defining phenotype of the tumour, and hence being presumably integral to the tumour type, (ii) present ab initio in only some cases of the tumour type, and presumably being regularly associated with, but incidental to, the essential features of the tumour type, (iii) occurring later in the course of the disease and thus being possibly a manifestation of in vivo genetic instability and "tumour progression" and (iv) arising probably by genetic instability, during the processes, especially cell culture, associated with ex vivo investigations. Genomic models are described which might account for the origin of these phenomena in each of these circumstances. PMID- 15810979 TI - Upregulation of MMP-9/TIMP-1 enzymatic system in eosinophilic meningitis caused by Angiostrongylus cantonensis. AB - Proteolysis depends on the balance between the proteases and their inhibitors. Matrix metalloproteinase-9 (MMP-9) and its specific inhibitors, tissue inhibitors of metalloproteinases (TIMP), contribute to eosinophilic inflammatory reaction in the subarachnoid space of the Angiostrongylus cantonensis-infected mice. The expression of MMP-9 in cerebrospinal fluid (CSF) was significantly increased in mice with eosinophilic meningitis, compared to that in uninfected ones. However, the TIMP-1 levels were unchanged and remained at basal levels at all time points, even in uninfected mice. Elevated MMP-9 mRNA expression coincided with protein levels and proteolytic activity, as demonstrated by means of positive immunoreactivity and gelatin zymography. CSF protein contents correlated significantly with MMP-9 intensity and CSF eosinophilia. In addition, immunohistochemistry demonstrated MMP-9 and TIMP-1 localization in eosinophils and macrophages. When the specific MMP inhibitor, GM6001, was added, MMP-9 enzyme activity was reduced by 45.4%. The percentage of eosinophil increased significantly upon the establishment of infection, but subsided upon inhibition. These results show that MMP-9/TIMP-1 imbalance in angiostrongyliasis may be associated with eosinophilic meningitis. PMID- 15810980 TI - A transmission electron microscopic study of microglia/macrophages in the hippocampal cortex and neocortex following chronic exposure to valproate. AB - In chronic administration of sodium valproate to rats, significant disorders of structural integrity of the hippocampal gyrus and the neocortex of the temporal lobe, observed in the last two stages of the experiment (after 9 and 12 months), coexisted with increased number of microglial cells and, especially after 12 months, with intense phagocytic activity within these cells. At the ultrastructural level, phagocyte microglial cells were hypertrophied with several broadened processes. Their cytoplasm contained rich lysosomal apparatus, numerous lipofuscin-like structures, lipid droplets and multilaminated bodies. The nuclei of these cells were characteristic oval or round and sometimes triangle in shape with dense and highly clumped heterochromatin, distinctly accumulated under nuclear envelope, and sparse euchromatin. Microglia/macrophages were frequently present in a close vicinity of changed neuronal somata and also close to the altered elements of the neuropil pyramidal layer of the cortex. Microglial response may, together with abnormalities in neurones, astroglia and blood-brain barrier, play a significant role in the development of experimental valproate encephalopathy. PMID- 15810981 TI - Increased thyroidal fat and goitrous hypothyroidism induced by interferon-gamma. AB - Summary Hashimoto's thyroiditis is associated with a diffuse lymphocytic infiltration of the stroma and a production of several cytokines, such as interferon-gamma (IFN-gamma). We previously reported that transgenic mice expressing IFN-gamma under the control of the thyroglobulin promoter develop primary hypothyroidism. In order to determine the long-term changes induced by IFN-gamma in the thyroid gland, we analysed cross-sectionally 202 mice (96 transgenic mice and 106 controls) of 0-650 days of age. Multiple linear regression analysis showed that, after adjusting for age and sex, thyr-IFN-gamma transgenic mice were 14% (3 g) smaller (P < 0.0001) and had a 5- to 6-fold bigger thyroid (P < 0.0001) than wild-type littermates. Transgenic thyroids showed striking histopathological changes in follicles, thyrocytes and stroma. Follicles were enlarged, irregular and were lined by thickened, granular and oxyphilic thyrocytes. The stroma contained a moderate and diffuse mononuclear infiltrate- mainly composed of macrophages--and, interestingly, a clear increase in the content of fat. These findings indicate that, in addition to hypothyroidism, chronic exposure of the thyroid to IFN-gamma leads also to macrophage infiltration and subsequent adipocyte expansion, suggesting a link between inflammation and fat accumulation. PMID- 15810982 TI - Effects of neutrophil depletion in the local pathological alterations and muscle regeneration in mice injected with Bothrops jararaca snake venom. AB - In order to study the role of neutrophils in the acute local pathological alterations induced by Bothrops jararaca snake venom, and in the process of skeletal muscle regeneration that follows, an experimental model was developed in mice pretreated with either an anti-mouse granulocyte rat monoclonal immunoglobulin G, which induces a profound neutropenia, or an isotype-matched control antibody. B. jararaca venom induced prominent haemorrhage and oedema, but only a moderate myonecrosis. No significant differences were observed in the extent of local haemorrhage, oedema and myonecrosis between neutropenic and control mice, suggesting that neutrophils do not play a determinant role in the acute pathological alterations induced by B. jararaca venom in this experimental model. Moreover, no differences were observed in skeletal muscle regeneration between these two experimental groups. In both the cases, limited areas of myonecrosis were associated with a drastic damage to the microvasculature and a scarce inflammatory infiltrate, with the consequent lack of removal of necrotic debris during the first week, resulting in a poor regenerative response at this time interval. Subsequently, a similar regenerative process occurred in both groups, and by 30 days, necrotic areas were substituted by groups of small regenerating muscle fibres. It is suggested that the drastic effect exerted by B. jararaca venom in the microvasculature precludes an effective access of inflammatory cells to necrotic areas, thereby compromising an effective removal of necrotic debris; this explains the poor regenerative response observed during the first week and the fact that there were no differences between neutropenic and control mice. As neutropenia in this model lasted only 7 days, the successful regenerative process observed at 30 days is associated with revascularization of necrotic regions and with a successful removal by phagocytes of necrotic debris in both groups. PMID- 15810983 TI - Enhancement of the repair of meniscal wounds in the red-white zone (middle third) by the injection of bone marrow cells in canine animal model. AB - Bone marrow stem cells (BMSCs) can differentiate into several cells that participate in the healing of meniscal wounds. To test this hypothesis, we examined the effects of injected BMSCs on the healing of meniscal wounds. Autologous BMSCs from eight adult dogs were injected into meniscal wounds (knee joints). After 12 weeks, the healing process was clinically and immunomorphologically evaluated using: (i) histochemical stains (haematoxylin and eosin, Masson trichrome and periodic acid-Schiff) and (ii) immunoperoxidase staining methods (CD3, CD79a, CD68, CD31 and alpha smooth-muscle actin for T, B lymphocytes, macrophages, endothelial cells and smooth-muscle lineage). Complete (six vs. three), partial (one vs. one) and no healing (one vs. four animals) of the meniscal wounds were observed in the injected and noninjected menisci. As compared with the noninjected menisci, examination of the tissues from the injected ones revealed: (i) marked angiogenesis (microvessel density: 3.22 +/- 0.66 vs. 6.50 +/- 2.10); (ii) chondrogenesis; (iii) prominent immune cell infiltrate (4.07 +/- 0.78 vs. 9.56 +/- 1.69, 8.33 +/- 0.77 vs. 3.67 +/- 1.00 and 4.38 +/- 0.62 vs. 11.1 +/- 1.43 for the total numbers of immune cells, lymphocytes and macrophages, respectively); and (iv) proliferation of the fibroblasts with marked deposition of collagen fibres (2.0 +/- 0.84 vs. 2.66 +/- 0.48). These values were statistically significantly higher for the injected menisci as compared with the noninjected ones (P >/= 0.05). Autologous BMSCs can improve meniscal wound healing. Whether this improvement occurs through BMSC differentiation into cells operational in the repair process, the release of certain mediator or other unknown mechanisms mandates further investigations. PMID- 15810984 TI - Differences in the effects of age on intestinal proliferation, crypt fission and apoptosis on the small intestine and the colon of the rat. AB - The increase in gastrointestinal epithelial tissue mass and the development of the gut can occur through three main mechanisms, namely elevated cell production from the intestinal crypts, by raised crypt number, which occurs through the process of crypt fission or by altered apoptosis. The small bowel and the colon have various rates of these, which were studied in rats of various ages. Wistar rats were fed ad libitum, and were killed at 3, 4, 6, 9, 12, 18, 26 and 48 weeks of age. Tissue was later stained and microdissected and the number of native mitoses and apoptotic figures per crypt and the percentage of crypts in fission were determined. There was an almost linear increase in body weight from 3 to 9 weeks, followed by a more gradual rise until 18 weeks. The weight of the stomach and the small intestine reached maximum values at 9 weeks, whereas the caecum and the colon approached this at 12 weeks. Mitotic activity per crypt in the small intestine increased from 3.8 +/- 0.1 at 3 weeks to 7.8 +/- 0.4 mitoses per crypt (P < 0.001) at 9 weeks and then decreased slightly; crypt fission increased from 4.6% +/- 0.8 at 3 weeks to 8.4 +/- 0.9% at 6 weeks and then decreased gradually reaching a value of 1.5 +/- 0.4% at 48 weeks. Apoptosis also peaked at 6 weeks and was then very low. In the colon, the proliferation decreased from 4.2 +/- 0.2 mitoses per crypt in the young (3 weeks) rat and reached a plateau by 9 weeks (2.5 +/- 0.1 mitoses per crypt, P < 0.001). Crypt fission also declined rapidly in the first 9 weeks (from 67.6 +/- 4.2 to 23.1 +/- 4.6%, P < 0.01) and then continued to decline, although at a lower rate. The crypt fission index at 48 weeks was 9.8 +/- 1.0. Apoptosis in the colon persisted throughout the duration of the study, 0.19 +/- 0.06 apoptotic bodies per crypt were seen at week 48. The development of the small intestine is more dependent on cell proliferation, whereas in the colon crypt fission is far more predominant, with the colon having fission indices approximately six times greater than those of the small intestine. Proliferative activity in the colon was approximately half that of the small intestine. PMID- 15810985 TI - Soluble bio-markers in vascular disease: much more than gauges of disease? AB - 1. In recent years demonstration of a direct association between slightly elevated serum levels of soluble proteins including the acute phase response proteins, selectins and intercellular adhesion molecules and the risk of developing vascular disease have been widely reported. These studies may provide the clinician with an insight into disease diagnosis, prognosis and disease activity. 2. The simplest interpretation of this data is that soluble proteins are just sensitive markers of inflammation. However, they may in fact be modulating inflammation directly through interaction with circulating cells. 3. Recent work has shown that these soluble proteins do indeed remain active and can bind to functional ligands expressed by circulating leucocytes. The current review focuses on the soluble proteins C-reactive protein and soluble P-selectin and describes previous studies characterizing their interaction with immune cells to modulate the pathogenesis of vascular disease. 4. The current review focuses on the soluble proteins C-reactive protein and soluble P-selectin and describes previous studies characterizing their interaction with immune cells to modulate the pathogenesis of vascular disease. PMID- 15810986 TI - Differential regulation of angiotensin II receptors during renal injury and compensatory hypertrophy in the rat. AB - 1. The renin-angiotensin system may be involved in the compensatory adaptations occurring after the reduction of renal mass and during the consecutive changes leading to chronic renal failure. We therefore investigated the regulation of angiotensin II receptors in two models of renal hypertrophy in the rat: hypertrophy following uninephrectomy (UNx) or subtotal nephrectomy (STNx). The level of angiotensin type 1 (AT1A-R and AT1B-R) and type 2 (AT2-R) receptor mRNA was quantified by competitive reverse transcription-polymerase chain reaction (RT PCR) in specific renal zones and the intrarenal distribution of angiotensin II receptors was analysed by immunohistochemistry. 2. In the UNx rats, AT1-R mRNA expression was not modified in the cortex or in the inner stripe of the outer medulla of the residual kidney at any time after the surgery (1, 4 and 12 weeks). In contrast, AT1-R mRNA expression was significantly reduced in these zones in STNx rats (-33% and -40%, respectively). This downregulation was organ-specific, as AT1-R mRNA levels were not modified in the liver. The proportions of AT1-R subtype (AT1A and AT1B) mRNA were unchanged by UNx or STNx. Very low levels of AT2-R mRNA were found in the cortex of all groups. Immunostaining revealed a similar localization of AT1-R in mesangial cells, proximal tubule, basolateral membrane of thick ascending limb, in both models of hypertrophy. AT1-R labelling was also detected in the apical membrane of intercalated cells of cortical collecting ducts. 3. This differential mRNA expression of angiotensin II receptors during compensatory hypertrophy and renal injury suggests that the development of renal hypertrophy is independent of AT1-R and AT2-R gene expression levels. PMID- 15810987 TI - Cardiovascular adaptations in rats submitted to a resistance-training model. AB - 1. The present study sought to evaluate cardiovascular adaptations, such as blood pressure (BP), heart rate (HR) and cardiac hypertrophy, to resistance training (RT) in a rat model. 2. The training protocol consisted of four sets of 10-12 repetitions of the squat exercise performed at 65-75% of one repetition maximum (1RM) over 4 weeks. Animals were randomly divided into three groups: control (n = 8, CO), electrically stimulated (n = 8, ES) and trained (n = 8, TR; also electrically stimulated). Blood pressure and HR were measured by a direct method in conscious rats after the training period. 3. All groups began with similar 1RM and 1RM/bodyweight (BW) ratio, however, at the end of the protocol only the TR group was different from the beginning (56% and 50%, respectively; both P < 0.01). The CO and ES groups had similar values for cardiac chambers weight/BW ratio, HR and diastolic, systolic and mean BP. Left ventricular hypertrophy (LVH) determined by the left ventricle (LV) weight/BW ratio was increased in the TR group (12%) when compared to CO (P < 0.01) or ES groups (P < 0.01). No changes were found in the weights of the atrium or right ventricle. Diastolic (14%) and mean BP (13%) were lower in the TR group (P < 0.05), whereas systolic BP and HR remained unchanged. 4. Collectively these results demonstrate that the rat RT model used is associated with significant development of cardiac hypertrophy and lowering of resting BP. These cardiovascular adaptations seem to a result of the training exercise and not influenced by stress since circulating catecholamine levels and adrenal gland weights remained unchanged in all groups. PMID- 15810988 TI - Renal denervation causes chronic hypotension in rats: role of beta1-adrenoceptor activity. AB - 1. Renal denervation (RDNX) chronically lowers mean arterial pressure (MAP) in normal rats but mechanisms leading to this hypotensive response remain unknown. 2. We hypothesized that this sustained decrease in arterial pressure was because of a loss of beta1-adrenoceptor mediated renin secretion. Male Sprague-Dawley rats were assigned to sham (SHAM; n = 9), unilateral (UniRDNX; n = 9), or bilateral (RDNX; n = 10) renal denervation groups and instrumented for telemetric MAP measurements, plasma renin concentration (PRC) measurements and intravenous infusion. Twenty-four h MAP, heart rate, sodium and water balances were recorded 5 days before, 3 days during and 3 days after 1-adrenoceptor blockade with atenolol. 3. The 5-day control MAP was significantly lower in RDNX (97 +/- 1 mmHg) compared to SHAM (105 +/- 2 mmHg) and UniRDNX (102 +/- 2 mmHg) rats. No significant differences in basal PRC were observed between RDNX (2.2 +/- 0.3 ngAng1/mL per h), UniRDNX (2.6 +/- 0.4 ng/Ang1/mL per h) and SHAM (2.6 +/- 0.4 ngAng1/mL per h) rats. By day 1 of atenolol, PRC was significantly lower in UniRDNX rats (1.8 +/- 0.2 ngAg1/mL per h) compared to control values, but was unchanged during atenolol infusion in the other groups. By day 3 of atenolol, MAP was significantly decreased in all groups, but the absolute levels of MAP remained statistically different between RDNX (87 +/- 1 mmHg) and SHAM (91 +/- 1 mmHg) groups. 4. We conclude that the arterial pressure lowering effect of RDNX is not solely dependent on the loss of neural control of renin release. PMID- 15810989 TI - Alpha1-receptor or adenosine A1-receptor dependent pathway alone is not sufficient but summation of these pathways is required to achieve an ischaemic preconditioning effect in rabbits. AB - 1. It is reported that alpha1-receptors and adenosine A1-receptors are involved in the ischaemic preconditioning (PC) effect on infarct size (IS). However, it is still unclear to what extent alpha1-receptors and adenosine A1-receptors contribute to the mechanism of PC. Therefore, we investigated the extent of the contribution of alpha1-receptors and adenosine A1 receptors to the PC effect on IS and examined the relationship between these receptors and protein kinase C. 2. Infarct size was measured in rabbits subjected to 30 min ischaemia and 48 h reperfusion. Tyramine (Tyr) was intravenously administered before 30 min ischaemia in the absence or presence of bunazosin (BN, alpha1-receptor blocker) and staurosporine (ST), a protein kinase C inhibitor, respectively. R(-)N6-(2 phenylisapropyl)-adenosine (PIA), a selective adenosine A1 agonist, was intravenously administered before 30 min ischaemia in the absence or presence of 8-p-sulphophenyltheophylline (8SPT), an adenosine blocker and ST, respectively. In the PC groups, BN, BN + PIA, 8SPT, 8SPT + Tyr or placebo saline was injected before or during PC. 3. Both Tyr and PIA reduced the IS, which was blocked by BN and 8SPT, respectively. The IS-reducing effect of Tyr or PIA was blocked by ST. The IS-reducing effect of PC was completely blocked by BN and 8SPT, respectively. The blocking effect of BN on the IS-reducing effect of PC was abolished by adding PIA during PC ischaemia. The blocking effect of 8SPT on the IS-reducing effect of PC was abolished by adding Tyr before PC ischaemia. 4. These data suggest that an alpha1-receptor dependent pathway exists and an adenosine A1-receptor dependent pathway, stimulation of both of which activates protein kinase C, then reduces the IS. However, exclusive stimulation of a single alpha1-receptor dependent pathway or a single adenosine A1-receptor dependent pathway alone is not sufficient but the summation of these pathways is required to achieve a PC effect on IS in rabbits. PMID- 15810990 TI - Withdrawal of intravenous glyceryl trinitrate: absence of rebound phenomena with transition to oral isosorbide dinitrate. AB - 1. Glyceryl trinitrate (GTN) is frequently infused intravenously as a component of the management of acute coronary syndromes (ACS). Abrupt cessation of GTN infusion after periods of more than 24 h administration often induces rebound vasoconstriction reflecting 'pseudotolerance'; this is also the basis of the 'zero hour phenomenon' during chronic nitrate therapy. The efficacy of oral nitrate regimens to prevent vasoconstriction following cessation of intravenous GTN has not been previously examined. Therefore, we investigated the effects of transition from intravenous GTN to oral isosorbide dinitrate (ISDN) on a parameter of apparent arterial stiffness in patients with ACS. 2. The effects of GTN infusion at 5 microg/min on augmentation index (AIx) were quantified in patients (n = 10) with stable angina pectoris in order to establish the magnitude of effect on apparent arterial stiffness. 3. This infusion rate of GTN reduced AIx from 23 +/- 10% (SD) to 3 +/- 14% (SD) (P < 0.01). The effect of transition from GTN infusion of greater than 24 h duration to ISDN (10 mg tds) were examined in patients (n = 16) with ACS (unstable angina/non-Q-wave myocardial infarction). No patient developed recurrent angina during the 24 h following cessation of GTN infusion. The level of AIx was 8 +/- 4% (SD) prior to GTN cessation and fell to 5 +/- 6% (SD) on ISDN (P = 0.05). 4. Thus, in patients treated for ACS, transition from intravenous GTN to low dose oral ISDN is associated with an incremental vasodilatation and no evidence of 'rebound' ischaemia. PMID- 15810991 TI - Effect of zinc sulphate and zinc methionine on growth, plasma growth hormone concentration, growth hormone receptor and insulin-like growth factor-I gene expression in mice. AB - 1. The current experiment was conducted to investigate the effect of zinc sulphate (ZnSO4) and zinc methionine (Zn-Met) on growth and their effect on plasma growth hormone (GH) concentration, growth hormone receptor (GHR) and insulin-like growth factor I (IGF-I) mRNA expression in mice. 2. Ninety male KunMing (KM) mice were randomly divided into three treatments. The control group was fed on a basal diet containing 11.67 mg/kg of zinc. The ZnSO4 group and Zn Met group were fed on the diets supplemented with ZnSO4 or Zn-Met at 30 mg/kg (containing zinc of 40.05 and 40.75 mg/kg, respectively). The mice were offered the test diets for 10 days. Weight gains and food intake were measured at the end of the experiment, zinc contents in liver and serum were determined using atomic absorption spectrophotometry; GH was determined by radioimmunoassay, the levels of GHR and IGF-I mRNA were determined with reverse transcript polymerase chain reaction. 3. Both ZnSO4 and Zn-Met enhanced weight gain and food intake in the mice, Zn-Met improved the growth and food intake more effectively than ZnSO4 did (P < 0.05). The both forms of zinc had no effect on GH and the level of GHR mRNA expression (P > 0.05) and they up-regulated the expression of IGF-I mRNA (P < 0.05). As compared to ZnSO4, Zn-Met enhanced the level of IGF-I mRNA significantly (P < 0.05). 4. Both ZnSO4 and Zn-Met had no effect on plasma GH and the expression of GHR mRNA, but they enhanced the expression of IGF-I mRNA. Zinc methionine enhanced the weight gain and up-regulated IGF-I mRNA expression more effectively than ZnSO4. PMID- 15810992 TI - Resistance to excessive bodyweight gain in risperidone-injected rats. AB - 1. The present study was carried out to explain the resistance of rats injected subcutaneously with risperidone, the atypical antipsychotic drug, for 21 consecutive days at 0.1 mg/kg per day (a dose equivalent to the one used for patients) to result in an excessive bodyweight despite the increase in diet uptake in rats against risperidone-induced decrease in body temperature. 2. Rectal temperature measurements were made in 8-week-old male Sprague-Dawley rats maintained under standard laboratory conditions using a 12 h daylight cycle. A s.c. injection of risperidone (0.05 mg/kg) produced hypothermia in rats, which was observed during the daily injection for 21 consecutive days. 3. Sera, white and brown adipose tissues, skeletal muscle and liver were extracted from 8-week old male Sprague-Dawley rats injected subcutaneously with risperidone (0.01 or 0.1 mg/kg per day) or a vehicle for 21 consecutive days. Serum levels of lipids, ketones and thyroid hormone were measured. The mRNA expression levels in these tissues and organs of the genes encoding the substances involved in heat production and/or lipid metabolism were investigated by using quantitative real time polymerase chain reaction amplification. 4. Serum nonesterified fatty acid levels in risperidone 0.1 mg/kg per day s.c. injected rats were significantly lower than those in vehicle-injected ones. Serum beta-hydroxybutyrate levels in risperidone-injected rats tended to decrease compared with those in vehicle injected ones. The serum level of neither triiodothyronine nor thyroxine was affected by risperidone s.c. injection at the doses examined, although their values were within normal limits. 5. Risperidone injection (0.1 mg/kg per day) for 21 consecutive days upregulated mRNA expressions in white adipose tissue of uncoupling protein 3 which dissipates energy as heat; peroxisome proliferator activated receptor (PPAR) gamma coactivator 1alpha which activates mitochondrial biogenesis to expand the oxidative machinery; and PPARalpha which is necessary for the fat-depletion of adipocytes for thermogenesis. The mRNA of lipogenic enzymes (acetyl-CoA carboxylase alpha, fatty-acid synthase and glycerol-3 phosphate acyltransferase), hormone sensitive lipase and beta1-adrenoceptor were also enhanced in white adipose tissue by the injection of 0.1 mg/kg per day risperidone. 6. These findings suggest that the materials for heat generation in white adipose tissue would be readily supplied, which in turn would reduce a storage of lipids in white adipose tissue resulting in the lower rate of bodyweight gain of rats. PMID- 15810994 TI - Ganglion blockade does not prevent cortisol-induced hypertension in man. AB - 1. The aim of the present study was to assess the effect of ganglion blockade on blood pressure in cortisol treated human subjects. 2. Four healthy male subjects were treated with cortisol 80 mg/day for a 5-day period. Ganglion blockade was achieved by intravenous trimethaphan. 3. Ganglion blockade did not significantly alter blood pressure in the pretreatment phase or on the last day of cortisol treatment. 4. Taken together with our previous observations that sympathetic activity is unaltered or reduced by cortisol, these results suggest that cortisol induced hypertension in humans is not a result of overactivity of the autonomic nervous system. PMID- 15810993 TI - Anaphylactic hepatic venoconstriction is attenuated by nitric oxide released via shear stress-dependent and -independent mechanisms in Guinea pig. AB - 1. The role of shear stress in nitric oxide (NO)-mediated attenuation of anaphylactic venoconstriction was studied using an isolated ovalbumin-sensitized guinea pig liver. 2. Guinea pigs were actively sensitized by a subcutaneous injection of 1 mg ovalbumin. Two weeks after sensitization, the livers were perfused with diluted blood under constant flow or constant perfusion pressure. The constant flow could result in increased shear stress during constriction, while the constant perfusion pressure could prevent changes in shear stress. Using the double occlusion technique to estimate the hepatic sinusoidal pressure, pre- and postsinusoidal constriction was evaluated. Hepatic anaphylaxis was induced by an injection of ovalbumin (4 microg) into the perfusate, the volume of which was 40 mL. 3. Under either constant flow or pressure, anaphylaxis caused venoconstriction of predominantly presinusoids over postsinusoids, although anaphylactic venoconstriction under constant pressure was significantly greater than that under constant flow. When shear stress was held constant by maintaining constant perfusion pressure, a NO synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME, 100 micromol/L), potentiated similarly both pre- and postsinusoidal constriction induced by anaphylaxis. This suggests that hepatic anaphylaxis shear stress-independently generates NO, resulting in dilatation of both pre- and postsinusoidal vessels in a similar magnitude. In contrast, when shear stress was allowed to rise under constant flow, anaphylactic presinusoidal constriction was preferentially potentiated by L-NAME. 4. Hepatic anaphylaxis can increase NO production in a shear stress-independent manner and dilates similarly both pre- and postsinusoids, while NO produced in a shear stress-dependent manner attenuates predominantly venoconstriction of the presinusoids where shear stress is preferentially increased. PMID- 15810995 TI - Optical imaging of respiratory neuron activity from the dorsal view of the lower brainstem. AB - 1. We visualized respiratory-related neuron network activity in the dorsal part of the pons and medulla of an in vitro preparation from newborn rats by optical recordings using a voltage-sensitive dye. We measured optical signals from several seconds before to several seconds after the inspiratory phase using the inspiratory motor nerve discharge as the trigger signal and we averaged the optical signals of 20-50 respiratory cycles to obtain an optical image correlating specifically to inspiratory activity. 2. Four areas that were excited or inhibited corresponding to the respiratory cycles were detected. (i) The most rostral activity was in the rostral and lateral parts of the pons, with activity mainly in the inspiratory phase, corresponding to the pontine-respiratory group. (ii) In the midpontine level, inspiratory activity followed by long-lasting hyperpolarization appeared in the midlateral parts. This part was presumed to reflect activity in the locus coeruleus. The hyperpolarization became almost negligible after treatment with the alpha-adrenergic antagonist, phentolamine. (iii) In the dorsal medulla, the predominantly inspiratory activity was detected at the rostral level of the area postrema. This part was considered to reflect activity mainly of the hypoglossal nucleus. (iv) At a similar level, we also detected weak and disperse inspiratory activity extending more laterally and caudally than that of the hypoglossal nucleus activity. This might reflect activity of the dorsal respiratory group. 3. In conclusion, the present optical recording study revealed that the dorsal part of the lower brainstem in the in vitro preparation is noticeably active as well as the ventral part shown in the previous study. This method is very useful for analysis of pharmacological properties, as well as the spatio-temporal pattern of respiratory-related network activity in the brainstem. PMID- 15810996 TI - Active role for the vasculature in the delivery of insulin to skeletal muscle. AB - 1. In the 80+ years since insulin's discovery, an enormous amount of literature has accumulated relating to its actions on body fat, glucose and protein metabolism. In particular, skeletal muscle has been extensively studied because of its major role as a site of insulin-mediated glucose disposal. Liver and adipose tissue are two other extensively studied sites of insulin action. Much less investigation has been directed towards delineating insulin's actions on cells other than myocytes, adipocytes and hepatocytes. 2. Over the past 5-10 years it has become increasingly evident that insulin exerts important actions on vascular cells. Here, we review evidence that insulin's action within muscle may be very much regulated by its ability to transit the vasculature to access the interstitial fluid (and hence the myocyte insulin receptor). Surprisingly little is known regarding the regulation of vascular events that first bring insulin to the capillary endothelium within muscle, whence presumably it transits from the vascular to the interstitial space. Recent studies suggest that insulin can increase blood flow and also influence the distribution of blood flow within skeletal muscle, potentially therefore regulating its own delivery to the capillary endothelium. Beyond insulin's ability to access the vascular lumen within skeletal muscle microvasculature lies the issue of its passing the endothelial barrier. Even less is known about the processes involved in insulin's actual transit across the endothelium. Available data do not clearly indicate whether this is a saturable, receptor-mediated process or a passive-diffusion pathway. Also, whether insulin in any manner regulates its own transit across the endothelium or its clearance via the lymphatic system is entirely unknown. 3. The aim of the present review is to identify areas where knowledge is deficient and highlight hypotheses which may lead to a better understanding of the coordinated relationship between insulin's vascular actions within muscle and its metabolic actions in that tissue. Even so, there is now sufficient evidence to indicate that insulin's vascular action within skeletal muscle is a major regulatory locus for its insulin mediated glucose disposal. PMID- 15810997 TI - Use of GLUT-4 null mice to study skeletal muscle glucose uptake. AB - 1. The present review focuses on the effects of varying levels of GLUT-4, the insulin-sensitive glucose transporter, on insulin sensitivity and whole body glucose homeostasis. 2. Three mouse models are discussed including myosin light chain (MLC)-GLUT-4 mice which overexpress GLUT-4 specifically in skeletal muscle, GLUT-4 null mice which express no GLUT-4 and the MLC-GLUT-4 null mice which express GLUT-4 only in skeletal muscle. Overexpressing GLUT-4 specifically in the skeletal muscle results in increased insulin sensitivity in the MLC-GLUT-4 mice. In contrast, the GLUT-4 null mice exhibit insulin intolerance accompanied by abnormalities in glucose and lipid metabolism. Restoring GLUT-4 expression in skeletal muscle in the MLC-GLUT-4 null mice results in normal glucose metabolism but continued abnormal lipid metabolism. 3. The results of experiments using these mouse models demonstrates that modifying the expression of GLUT-4 profoundly affects whole body insulin action and consequently glucose and lipid metabolism. PMID- 15810998 TI - Glucose phosphorylation as a barrier to muscle glucose uptake. AB - 1. Glucose phosphorylation is the first irreversible step of the muscle glucose uptake pathway and is catalysed by a hexokinase isozyme. 2. While glucose transport is the primary barrier to muscle glucose uptake during basal conditions, glucose phosphorylation becomes an important barrier to muscle glucose uptake during stimulated conditions such as hyperinsulinaemia or exercise. 3. High fat feeding markedly impairs insulin- and exercise-stimulated muscle glucose uptake. As hexokinase II overexpression corrects this dietary induced deficit during exercise, glucose phosphorylation is a site of impairment following high fat feeding. 4. Exercise is an important tool for diagnosing deficits in glucose phosphorylation. PMID- 15810999 TI - Interaction of physiological mechanisms in control of muscle glucose uptake. AB - 1. Control of glucose uptake is distributed between three steps. These are the rate that glucose is delivered to cells, the rate of transport into cells, and the rate that glucose is phosphorylated within these same cells. The functional limitations to each one of these individual steps has been difficult to assess because they are so closely coupled to each other. Studies have been performed in recent years using complex isotopic techniques or transgenic mouse models to shed new light on the role that each step plays in overall control of muscle glucose uptake. 2. Membrane glucose transport is a major barrier and glucose delivery and glucose phosphorylation are minor barriers to muscle glucose uptake in the fasted, sedentary state. GLUT-4 is translocated to the muscle membrane during exercise and insulin-stimulation. The result of this is that it can become so permeable to glucose that it is only a minor barrier to glucose uptake. 3. In addition to increasing glucose transport, exercise and insulin-stimulation also increase muscle blood flow and capillary recruitment. This effectively increases muscle glucose delivery and by doing so, works to enhance muscle glucose uptake. 4. There is a growing body of data that suggests that insulin resistance to muscle glucose uptake can be because of impairments in any one or more of the three steps that comprise the process. PMID- 15811000 TI - Comments on journal guidelines for reporting statistics. PMID- 15811001 TI - The molecular basis of variation in human color vision. AB - Common variation in red-green color vision exists among both normal and color deficient subjects. Differences at amino acids involved in tuning the spectra of the red and green cone pigments account for the majority of this variation. One source of variation is the very common Ser180Ala polymorphism that accounts for two spectrally different red pigments and that plays an important role in variation in normal color vision as well as in determining the severity of defective color vision. This polymorphism most likely resulted from gene conversion by the green-pigment gene. Another common source of variation is the existence of several types of red/green pigment chimeras with different spectral properties. The red and green-pigment genes are arranged in a head-to-tail tandem array on the X-chromosome with one red-pigment gene followed by one or more green pigment genes. The high homology between these genes has predisposed the locus to relatively common unequal recombination events that give rise to red/green hybrid genes and to deletion of the green-pigment genes. Such events constitute the most common cause of red-green color vision defects. Only the first two pigment genes of the red/green array are expressed in the retina and therefore contribute to the color vision phenotype. The severity of red-green color vision defects is inversely proportional to the difference between the wavelengths of maximal absorption of the photopigments encoded by the first two genes of the array. Women who are heterozygous for red and green pigment genes that encode three spectrally distinct photopigments have the potential for enhanced color vision. PMID- 15811002 TI - Is osseous dysplasia a primary feature of neurofibromatosis 1 (NF1)? AB - Characteristic skeletal lesions are a cardinal feature of the autosomal dominant condition, neurofibromatosis 1 (NF1). The most frequently involved skeletal sites are the sphenoid wing, vertebrae, and tibia. Osseous lesions may range in severity in NF1 but are often progressive. They may lead to serious clinical consequences and be resistant to treatment. The skeletal lesions of NF1 are usually considered to be 'dysplasias', i.e. primary defects of bone, although there is no direct evidence supporting this interpretation. Moreover, it is difficult to understand why a generalized dysplasia of bone would produce focal lesions that show such a striking predisposition to only a few bones. We review the clinical and pathological features of NF1 skeletal lesions and propose that they result from an abnormal response of NF1 halpoinsufficient bone to abnormal mechanical forces rather than from a primary osseous dysplasia. PMID- 15811003 TI - Looping out links Rett syndrome with loss of imprinting error. PMID- 15811004 TI - TPH2: an answer for unipolar depression? PMID- 15811005 TI - Osteoarthritis? Try asporin. PMID- 15811006 TI - Calibration of 6q subtelomere deletions to define genotype/phenotype correlations. AB - Testing for subtelomere abnormalities in patients with idiopathic mental retardation has become a useful diagnostic tool. However, limited data exist regarding genotype/phenotype correlations for specific subtelomere imbalances. We have ascertained five patients with 6q subtelomere deletions either as a result of an isolated deletion or as a result of an unbalanced translocation, and developed a molecular ruler assay utilizing BAC or PAC clones and determined the size of the deleted regions to range from <0.5 to 8 Mb. To establish genotype/phenotype correlations for distal 6q, we compared the clinical features of these patients to previously reported cases of 6q subtelomere and cytogenetically visible deletions and found that they shared multiple abnormalities, suggesting that the causative genes may lie in the region of the smallest 6q subtelomeric deletion, approximately 400 kb from the telomere. However, multiple unique features were present only in patients with cytogenetically visible 6q deletions, indicative that genes involved in the development of these features may lie more proximally on 6q. These initial studies demonstrate the ability to develop genotype/phenotype correlations for subtelomere rearrangements, which will aid in the diagnosis and prognosis of these patients and may help narrow the search for relevant developmental genes. PMID- 15811007 TI - Reproductive decision making before and after predictive testing for Huntington's disease: an Australian perspective. AB - A retrospective study examined both pre- and post-result reproductive decision making for 281 people at risk for Huntington's disease aged 18-45 years who had undergone predictive testing in one centre in Australia between 1990 and 2002. Forty-eight per cent of subjects had one or more pre-result pregnancies, and of these, three had prenatal linkage testing. One high-risk (50%) pregnancy was terminated. Four couples chose an alternative reproductive option. Following testing, data were available for 231 subjects, and no significant difference was found between mutation carriers and non-carriers in the occurrence of post-result pregnancies. This contrasts with the finding of a recent European study, although the outcome of the present study may have been influenced by loss of follow-up data for 50 subjects. Five carriers (17%) had a total of six prenatal tests. Four showed a carrier result and these pregnancies were terminated. Two carriers utilized an alternative reproductive option (donor insemination and pre implantation genetic diagnosis). The results of this study confirm previous findings of a low uptake of prenatal testing and alternative reproductive options by people at risk for Huntington's disease undergoing predictive testing. PMID- 15811008 TI - Evidence for, and a spectrum of, neurological involvement in carriers of the fragile X pre-mutation: FXTAS and beyond. AB - A neurodegenerative disorder, fragile X-associated tremor/ataxia syndrome (FXTAS), occurs in some older men carrying a small CGG repeat expansion (pre mutation) in the FMR1 gene. We surveyed a sample of older pre-mutation males to estimate the prevalence and spectrum of neurological involvement. Twelve pre mutation males aged 50-82 years and 11 age-matched normal controls ascertained in an unbiased manner were included in a neurological assessment that also used standard scales for tremor (Clinical Rating Scale for Tremor), ataxia (International Cooperative Ataxia Rating Scale, ICARS) and parkinsonian signs (Unified Parkinson's Disease Rating Scale). Axial FLAIR images of the brain, and neuropsychological and molecular tests were also conducted in pre-mutation carriers. The neurological disorder meeting all the criteria for diagnosis of 'definite' to 'possible' FXTAS occurred in five of 12 pre-mutation carriers (41.7%), and this prevalence was significantly higher compared with normal controls (0%). The ataxia (ICARS) score and the sum of all three tremor/ataxia scores were significantly higher in pre-mutation carriers than in controls, and mRNA was elevated in all but one carrier, but did not correlate with the degree of neurological involvement. In conclusion, the findings provide further evidence that the pre-mutation allele of FMR1 is a significant cause of late-onset neurodegeneration, presenting with a broad spectrum of clinical manifestations. PMID- 15811009 TI - X-linked adrenoleukodystrophy in Spain. Identification of 26 novel mutations in the ABCD1 gene in 80 patients. Improvement of genetic counseling in 162 relative females. AB - In this study, we analyzed the ABCD1 gene in 80 X-linked adrenoleukodystrophy (X ALD) patients from 62 unrelated families. We identified 53 different mutations, of which 26 are novel and two are non-pathogenic sequence variants (L516L and 3'UTR, 2246C/G) that have been previously described. The Spanish population had significant allelic heterogeneity, in which most of the mutations were exclusive to a single family 47/53 (88.7%). Only six mutations (Y174S, G277R, FsE471, R518Q, P543L, and R554H) were found in more than one family. Mutations G277R, P543L, and R554H were the most frequent, each of them being found in three patients (5%). Intra-familiar phenotype variability was observed in most of the families, but in one, with the novel mutation R120P, only the adult mild phenotype was present (five hemizygous family members). We detected 80 heterozygous women by mutation analysis, but only 78 of them showed increased very-long-chain fatty acid levels. In conclusion, this study extends the spectrum of mutations in X-ALD and facilitates the identification of heterozygous females. Our results are also consistent with previous studies reporting the difficulty of predicting genotype-phenotype correlation. PMID- 15811010 TI - HJV gene mutations in European patients with juvenile hemochromatosis. AB - A large variety of mutations within the genes encoding hepcidin (HAMP) and hemojuvelin (HJV) have been identified in patients with the severe iron overload disorder juvenile hemochromatosis (JH). The aim of the present study was to evaluate the molecular background of JH in patients from central parts of Europe. Sequence analyses of HAMP and HJV were performed in seven JH patients from six families from Germany, Slovakia, and Croatia. For detection of the G320V mutation in HJV, a rapid polymerase chain reaction-based assay was developed. No mutations were found within the HAMP gene. Six of seven (86%) JH patients carried at least one copy of the G320V mutation within the HJV gene. Four of these patients were homozygous for the G320V mutation. In addition, two novel HJV mutations were identified (C119F and S328fsX337). Taken together, the present study demonstrates that molecular analysis of the HJV gene is a powerful tool for an early and reliable diagnosis of JH. As in affected patients from Greece, the G320V mutation seems to be widely distributed among JH patients from central parts of Europe. Therefore, detection of the G320V mutation could identify the majority of JH cases from these regions non-invasively. PMID- 15811011 TI - Molecular analysis of non-syndromic preaxial polydactyly: preaxial polydactyly type-IV and preaxial polydactyly type-I. AB - Human GLI3 gene mutations have been identified in several phenotypes of digital abnormality such as Greig cephalopolysyndactyly syndrome, Pallister-Hall syndrome, preaxial polydactyly type-IV (PPD-IV) and postaxial polydactyly. However, the different phenotypes resulting from GLI3 mutations have not yet been properly defined. We have experienced two types of digital abnormality without other complicating developmental defects; a family with foot PPD-IV with syndactyly of the third and fourth fingers, and four sporadic cases with biphalangeal thumb polydactyly (PPD-I). The genes responsible for syndactyly of the third and fourth fingers (syndactyly type-I) and PPD-I have not yet been identified; we therefore examined the involvement of the GLI3 gene in these subtypes of digital abnormality. We found a non-sense mutation in the GLI3 gene in the family with foot PPD-IV accompanied with hand syndactyly of the third and fourth fingers, but no mutations were detected in the GLI3 gene in the four other cases with PPD-I alone. Thus, the phenotype of foot PPD-IV accompanied with hand syndactyly of the third and fourth fingers may result from a GLI3 mutation, whereas the PPD-I phenotype alone is not caused by GLI3 gene defect. These results will help to define the phenotypic spectrum of GLI3 morphopathies, which have been recently proposed. PMID- 15811012 TI - Premature ovarian failure associated with a small terminal Xq deletion: narrowing the POF1 region down to Xq27.2/Xq27.3-qter. PMID- 15811013 TI - Pendred's syndrome and non-syndromic DFNB4 deafness associated with the homozygous T410M mutation in the SLC26A4 gene in siblings. PMID- 15811014 TI - X-linked myotubular myopathy: mutation R69C identified in a family with multiple neonatal deaths. PMID- 15811015 TI - Identification and molecular characterization of 18 novel mutations in the ATP7B gene from Indian Wilson disease patients: genotype. PMID- 15811016 TI - Skewed X-inactivation in a family with mental retardation and PQBP1 gene mutation. PMID- 15811017 TI - Mental retardation, keratoconus, febrile seizures and sinoatrial block: a previously undescribed autosomal recessive disorder. PMID- 15811019 TI - Introduction: the modern gladiatorial arena: Gram-positive pathogens versus new therapies. PMID- 15811020 TI - Biological counterstrike: antibiotic resistance mechanisms of Gram-positive cocci. AB - The development of antibiotic resistance by bacteria is an evolutionary inevitability, a convincing demonstration of their ability to adapt to adverse environmental conditions. Since the emergence of penicillinase-producing Staphylococcus aureus in the 1940s, staphylococci, enterococci and streptococci have proved themselves adept at developing or acquiring mechanisms that confer resistance to all clinically available antibacterial classes. The increasing problems of methicillin-resistant S. aureus and coagulase-negative staphylococci (MRSA and MRCoNS), glycopeptide-resistant enterococci and penicillin-resistant pneumococci in the 1980s, and recognition of glycopeptide-intermediate S. aureus in the 1990s and, most recently, of fully vancomycin-resistant isolates of S. aureus have emphasised our need for new anti-Gram-positive agents. Antibiotic resistance is one of the major public health concerns for the beginning of the 21st century. The pharmaceutical industry has responded with the development of oxazolidinones, lipopeptides, injectable streptogramins, ketolides, glycylcyclines, second-generation glycopeptides and novel fluoroquinolones. However, clinical use of these novel agents will cause new selective pressures and will continue to drive the development of resistance. This review describes the various antibiotic resistance mechanisms identified in isolates of staphylococci, enterococci and streptococci, including mechanisms of resistance to recently introduced anti-Gram-positive agents. PMID- 15811021 TI - Current and emerging serious Gram-positive infections. AB - Serious infections caused by Gram-positive pathogens are increasingly difficult to treat because of pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE) and penicillin-resistant Streptococcus pneumoniae. The more recent emergence of vancomycin-intermediate and -resistant MRSA (VISA and VRSA) has further compromised treatment options. Reports of resistance to, and clinical failures with newer antimicrobial agents such as linezolid and quinupristin/dalfopristin are also emerging. Consequently, there is a clinical need for new antimicrobial agents that have suitable pharmacokinetic properties and safety profiles, with activity against these Gram positive pathogens. PMID- 15811022 TI - Efficacy of current agents used in the treatment of Gram-positive infections and the consequences of resistance. AB - The proportion of pathogens causing hospital-onset infections that are resistant to antimicrobial agents continues to increase worldwide. Inadequate antimicrobial therapy is an important factor in the emergence of resistance and is associated with increased mortality. In the USA in 2000, the National Nosocomial Infections Surveillance system reported that >50% of Staphylococcus aureus isolates collected from intensive care units were resistant to methicillin (MRSA). The emergence of community-acquired MRSA is a new concern. MRSA are associated with adverse clinical outcomes and increased hospital costs. The increasing prevalence of MRSA contributes to the use of glycopeptides; however, isolates with intermediate and full resistance to vancomycin and teicoplanin are now being reported. Newer agents, such as the oxazolidinone linezolid, are effective in the treatment of serious Gram-positive infections; however, linezolid-resistant isolates of Enterococcus faecium, Enterococcus faecalis and S. aureus have been reported. Therefore, there is an unmet clinical need for new agents with activity against Gram-positive pathogens. Daptomycin, a lipopeptide with a novel mode of action, was recently approved for the treatment of skin and soft tissue infections in the USA. The two case studies presented herein detail experience with the use of daptomycin in the USA. PMID- 15811023 TI - An epidemic of occupational contact dermatitis from an acrylic glue. AB - Dermatological examinations were performed in 81 workers involved in the manufacture of electric coils for television displays, who had worked for 4 years in contact with a glue containing isobornyl acrylate, acrylic acid, N,N dimethyleneacrylamide, phosphine oxide, bis(2,6-dimethoxybenzoyl) (2,4,4 trimethylpentyl)- and beta-carboxyethyl acrylate. The glue was cured by 350-500 nm ultraviolet and visible radiations. Acrylate-specific dermal lesions were detected in 21 (25.9%) people. Occupational irritant contact dermatitis was diagnosed in 12 (15%) of the workers and occupational allergic contact dermatitis in 9 (11.2%). 12 people reacted to acrylates. Cross-reactions with methacrylates were not observed. The highest number of positive tests was obtained with triethyleneglycol diacrylate (10 people) and diethyleneglycol diacrylate (9), followed by 1,6-hexanediol diacrylate (5), 1,4-butanediol diacrylate (4), beta carboxyethyl acrylate (3), tripropyleneglycol diacrylate (2) and pentaerythritol triacrylate (2). No cases of allergy to isobornyl acrylate, N,N methylenebisacrylamide or phosphine oxide were noted. PMID- 15811024 TI - Airborne irritant contact dermatitis due to synthetic fibres from an air conditioning filter. AB - We describe 8 cases of occupational airborne irritant contact dermatitis in intensive care unit (ICU) employees caused by synthetic (polypropylene and polyethylene) fibres from an air-conditioning filter. Not until a workplace investigation was conducted, was it possible to clarify the unusual sequence of events. High filter pressure in the intensive care air-conditioning system, maintained to establish an outward airflow and prevent microorganisms from entering the ward, probably caused fibres from the filter to become airborne. Upon contact with air-exposed skin, fibres subsequently provoked skin irritation. Test periods in the ICU with varying filter pressures, in an attempt to improve environmental conditions, led to even higher filter pressure levels and more complaints. The sometimes-very-low humidity might have contributed to development of skin irritation. The fact that most patients recovered quickly after treatment with emollients and changing the filters made it most likely that the airborne dermatitis was of an irritant nature. PMID- 15811025 TI - Metal allergy in Singapore. AB - This is a clinical epidemiologic study to determine the frequency of metal allergy among patch-tested patients in the years 2001-2003. The results are compared with those of previous studies. All patients diagnosed as having allergic contact dermatitis in the National Skin Centre, Singapore, from January 2001 to December 2003 were studied retrospectively. The frequency of positive patch tests to the following metals were nickel 19.9%, chromate 5.6%, cobalt 8.2% and gold 8.3%. The frequency of nickel allergy has been steadily rising over the last 20 years. The most common sources of nickel allergy are costume jewelry, belt buckles, wrist watches and spectacle frames. After declining from 1984 to 1990, chromate and cobalt allergies have also been steadily increasing subsequently. The most common sources of chromate allergy were cement, leather and metal objects. Most positive patch tests to cobalt are regarded as co sensitization due to primary nickel or chromate allergies. There has been a steep increase in positive patch tests to gold from 2001 to 2003, which is difficult to explain because the relevance and sources of such positive patch tests can rarely be determined with certainty. There has been an overall rise in the frequency of metal allergy in the last 20 years. PMID- 15811026 TI - 7 cases of hydrofluoric acid burn in which calcium gluconate was effective for relief of severe pain. AB - We report 7 cases of chemical burns due to hydrofluoric acid (HF). The patients suffered from severe pain. However, the pain was relieved after treatment with calcium gluconate. 6 out of the 7 cases were men. At the accidental exposures, all the patients had been engaged in washing or cleaning work and received burns on their hands and/or fingers. In one case, the forearm was also involved. During such work, all the patients had used rubber gloves, but the gloves had pinholes. For the treatments, 4% calcium gluconate jelly was applied in 5 cases and 4 of 7 were subcutaneously injected with 8.5% calcium gluconate. The involved nails were removed in 5 cases. It is concluded that physicians should provide calcium gluconate jelly and subcutaneous injections to treat an HF burn and should not hesitate to remove the involved nails. To prevent chemical burn due to HF, education and reeducation of workers regarding the hazard of this chemical are necessary. PMID- 15811027 TI - Contact allergy to methyldibromo glutaronitrile--data from a 'front line' network. AB - The preservative methyldibromo glutaronitrile (MDBGN) has caused an epidemic of contact allergy in Europe. However, most data concerning contact allergy comes from hospital departments of dermatology. As a part of the primary health care sector, Danish dermatologists in private practice provide the front line of care for patients with skin diseases. Data from this source may therefore better reflect trends in the general population than material from hospital departments of dermatology. In this study, the frequency of MDBGN allergy and the characteristics of patients seen by dermatologists in private practice were studied. In 1 year, 2146 patients were patch tested by the participating dermatologists from 4 clinics in various parts of Denmark. 5% (110) had positive patch tests to MDBGN, with no difference between the sexes. After adjustment was made for other background variables, multivariate analysis using logistic regression showed a significant association between hand eczema and MDBGN allergy (OR 2.5, P < 0.001). In 53 cases (52.4%), the patch test reaction to MDBGN was judged to be of current relevance. Creams and lotions accounted for 31% of the identified causative products and liquid soaps for 23%. It is concluded that contact allergy to MDBGN is frequent among patients seen by dermatologists in private practice. This is consistent with results from hospital departments of dermatology in Europe and indicates a general trend and ongoing epidemic in the general population. The significant relationship between hand eczema and MDBGN allergy is of concern, and the finding that wash-off products, especially liquid soaps, play a significant role in MDBGN allergy calls for a revision of the safety assessment of MDBGN in wash-off products. PMID- 15811028 TI - Pustular psoriasis and the Kobner phenomenon caused by allergic contact dermatitis from zinc pyrithione-containing shampoo. AB - Zinc pyrithione is a shampoo ingredient that has been shown to be safe and effective for dandruff and scalp psoriasis. It is thought to decrease the cell turnover rate in hyperproliferative dermatoses such as psoriasis, and also has fungistatic and antimicrobial activity, although its exact mode of action is unknown. In psoriasis, external factors, such as trauma, infection and drugs, may provoke aggravated manifestations of psoriatic skin lesions. Rarely, irritant or allergic mechanisms are likely causes of psoriatic flare and Kobnerization. A patient had had stable psoriasis for 25 years and no any other skin disease. Within 20 days, she developed an aggravated scaly erythematous patch on the scalp, where a shampoo had been applied, and simultaneously developed pustular psoriasis on both forearms. Patch testing showed a relevant sensitization to zinc pyrithione, and we observed symptomatic aggravation by provocation testing with zinc pyrithione shampoo. We report a rare case of psoriasis aggravated by the induction of allergic contact dermatitis from zinc pyrithione after using antidandruff shampoo. PMID- 15811029 TI - 'Black henna' tattoos: an occult source of natural rubber latex allergy? AB - Temporary 'black henna' tattoos are an increasingly popular body decoration with a growing incidence of associated adverse events. We report the case of a 14 year old girl presenting with an acute allergic contact dermatitis to hair dyes. Some years previously she had a 'black henna' tattoo with prior application of a tacky transfer to the skin to outline the design followed some days later by an acute localised blistering reaction. Patch testing to an extended British Contact Dermatitis Society standard series showed relevant positive reactions to paraphenlyenediamine (PPD) and thiuram mix, as well as to several of the azo disperse dyes. We went on to perform prick testing to natural rubber latex (NRL), showing a positive reaction to the commercial HEP 100 concentration. We propose that through the initial acute reaction caused by PPD in the 'black henna' our patient was not only sensitized to PPD, but also to thiuram and natural rubber latex which may have been present in the tacky transfer applied prior to 'tattooing.' We would add that temporary 'black henna' tattoos may have more important consequences than previously thought on patients' future health by sensitization to NRL. PMID- 15811030 TI - Prognosis of contact dermatitis in epoxy resin workers. AB - Between January 1993 and February 2002, 40 of 1354 (3%) workers who attended a tertiary referral occupational dermatology clinic were diagnosed with allergic contact dermatitis (ACD) from epoxy resin systems (ER). 20 of these patients were followed up at least 2 years post-diagnosis to establish prognosis. Characteristics collected at diagnosis, including age, atopic status and severity, were compared between the follow-up and not followed-up groups. No significant differences were observed. A clinician contacted patients and administered a telephone questionnaire based on variables identified from the literature, which were considered of importance for prognosis. All patients reported improvement of their skin condition since diagnosis. 12 of the 20 {60%[95% confidence interval (CI) 56-94%]} patients had applied for workers' compensation; all of these claims were successful. 16 had ceased working with ER. Of these, 9 [56% (95% CI 29-80%)] reported complete healing and 7 [44% (95% CI 19 70%)] reported ongoing dermatitis. Although no conclusions could be drawn because of the small sample size, factors that may be associated with a poor prognosis were age, atopy, duration of symptoms and severity at diagnosis. The prognosis of ACD from ER is not always favourable, even if a worker ceases exposure. PMID- 15811031 TI - Allergic contact dermatitis in Iran. AB - The frequency of sensitization to contact allergens varies in different countries because of both genetic and, more importantly, allergen exposure variations. The objective is to determine the frequency of sensitization to contact allergens in Iranian patients with dermatitis. 250 patients with a clinical diagnosis of contact dermatitis and/or atopic dermatitis were evaluated with a 28-allergen screening series recommended by the German Contact Dermatitis Research Group from September 2002 to April 2004 in Tehran, Iran. The patches were applied on the back of the patients, removed after 24 hr and the readings were taken on 24 hr, 48 hr and 72 hr after application. 126 patients (50.4%) showed at least 1 positive reaction, and 23 patients (9.2%) had more than 2 positive reactions. 189 (84.4%) of 224 positive reactions had past and/or present clinical relevance. The 5 most common allergens were nickel sulfate 70 (28.0%), cobalt chloride 32 (12.8%), para-tertiarybutyl phenol formaldehyde resin 20 (8.0%), potassium dichromate 13 (5.2%) and colophony 13 (5.2%). Contact allergy to nickel sulfate was significantly more common in female patients and in patients under 40 years of age (P < 0.05). Nickel sulfate is the most common contact allergen in Iran, mostly affecting women and younger patients probably because of more exposure. PMID- 15811032 TI - Severe contact allergy to footwear in a young child. PMID- 15811033 TI - Contact allergy to cinnamal in a patient with oral lichen planus. PMID- 15811034 TI - A case of type IV hypersensitivity to topiramate and carbamazepine. PMID- 15811035 TI - Contact allergy to topical corticosteroids in children with atopic dermatitis. PMID- 15811036 TI - Allergic contact dermatitis from triphenyl phosphite. PMID- 15811037 TI - Erythema multiforme due to methotrexate reproduced with patch test. PMID- 15811038 TI - Garlic-fingered chefs. PMID- 15811039 TI - Occupational allergic contact dermatitis to sodium lauroyl sarcosinate in the liquid soap. PMID- 15811040 TI - Provocative use test of 1 euro coin in nickel-sensitized subjects. PMID- 15811041 TI - Contact urticaria and respiratory symptoms induced by indirect contact with goat dander: a case report on vet's wife exposure. PMID- 15811042 TI - Contact dermatitis by dipivefrine. PMID- 15811043 TI - Oxidative stress in mouse skin following application of contact allergenic 5 chloro-2-methyl-4-isothiazolin-3-one and oxazolone. PMID- 15811044 TI - Diagnostic quality of the patch test preparation monoethanolamine 2% pet. PMID- 15811045 TI - Occupational airborne contact dermatitis from sporadic exposure to tetrazepam during machine maintenance. PMID- 15811046 TI - Eyelid dermatitis with positive patch test to coconut diethanolamide. PMID- 15811047 TI - Effect of air-polishing devices on the gingiva: histologic study in the canine. AB - OBJECTIVES: The aim of this study was to evaluate histologically the immediate effect of two types of air-polishing devices (APDs) on the gingiva. MATERIAL AND METHODS: The buccal gingiva in four mongrel dogs was exposed to 5, 10 and 20 s of instrumentation, applying a hand-piece type of APD (HP-APD) PROPHYflex and a stand-alone type of APD (SA-APD) Jetpolisher in a split-mouth design. Immediately after treatment, the exposed gingiva was excised by sharp dissection and fixed in 10% buffered formalin. For each specimen, 15 sections, 20 microm apart, corresponding to the central part of the treated and control untreated gingiva were cut and stained with haematoxylin and eosin. Stained sections were examined histomorphometrically for keratin width and epithelial cell layer (ECL) (prickle and basal cell layers) width. Extent of erosion was expressed as loss of keratin and ECL compared with control. RESULTS: Microscopic examination presented changes in keratin and ECL, including keratin detachment and disruption of the normal ECL architecture. The erosive changes in the gingiva caused by both APD, positively correlated with instrumentation time (p<0.001) and type of instrument (p=0.008). Keratin loss was significantly higher for SA APD than for the HP APD in each time interval (p=0.019). Following exposure for 5 s, both APD caused a 25% loss of the ECL. Exposure for 10 and 20 s revealed a significantly greater ECL loss caused by the SA-APD than the HP-APD (p=0.018). Exposure for 20 s was the only time interval that presented the area of total epithelium erosion with SA-APD causing significantly more (p=0.002) areas of total epithelial erosion than the HP-APD. CONCLUSION: Gingiva exposure to air-polishing slurry delivered by APD caused localized trauma because of epithelial erosive changes with severity, positively correlated with instrumentation time and design principles of the applied APD. The clinician should be aware of the potential insult of the gingiva when applying the APD and careful precautions should be taken. PMID- 15811048 TI - In vivo antimicrobial effectiveness of an essential oil-containing mouth rinse 12 h after a single use and 14 days' use. AB - OBJECTIVES: Two studies were conducted to determine the antimicrobial effect of rinsing with an essential oil-containing mouth rinse 12 h after a single rinse and 12 h after 2 weeks of twice daily rinsing, during the daytime and overnight. MATERIALS AND METHODS: These studies utilized a randomized, double-blind, controlled crossover design. Following baseline sampling of bacteria from supragingival plaque and the dorsum of the tongue, subjects began twice-daily rinsing with either an essential oil mouth rinse containing 0.09% zinc chloride (Tartar Control Listerine Antiseptic) or a negative control rinse. Bacterial sampling was repeated 12 h after the first rinse, and again 12 h after the final rinse 14 days later. The sampling schedule was adjusted according to whether the study was investigating daytime or overnight activity. Samples were plated on Schaedlers medium (total anaerobes), Schaedlers Nalidixic/Vancomycin medium (Gram negative anaerobes), and OOPS medium (volatile sulphur compound (VSC)-producing organisms). Inter-group log10 transformed colony-forming units/ml counts from samples of supragingival plaque and tongue swabs on each of the three media were compared by analysis of covariance. RESULTS: The mean bacterial counts in subjects using the essential oil mouth rinse were significantly lower (p< or =0.005) than mean counts in subjects using the control rinse in all the comparisons, i.e., tongue and supragingival plaque samples on each of three media at two sampling periods in the daytime and overnight study, respectively. Mean bacterial count percent reductions for plaque samples ranged from 56.3 to 95.3; percent reductions for tongue samples ranged from 61.1 to 96.1. There was a trend to higher reductions after 14 days' rinsing than after the initial rinse. CONCLUSION: Rinsing with the essential oil mouth rinse can have long-lasting effects in reducing anaerobic bacteria overall as well as Gram-negative anaerobes and VSC-producing bacteria. The significant reductions in numbers of these bacteria produced by the essential oil mouth rinse, both in plaque and on the dorsum of the tongue, can play a key role in explaining the essential oil mouth rinse's effectiveness in reducing supragingival plaque and gingivitis as well as its effectiveness in controlling intrinsic oral malodor over prolonged periods. PMID- 15811049 TI - Efficacy of Listerine Antiseptic in reducing viral contamination of saliva. AB - AIM: The anti-viral efficacy of oral antimicrobial rinses has not been adequately studied in terms of potential clinical significance. As a follow-up to an in vitro study on the effect of oral antiseptics on Herpes simplex virus, Type 1, this study was undertaken to evaluate the in vivo effect of an essential oil containing oral antiseptic on the reduction of viral titer in saliva during active viral infection. METHOD: Patients were recruited and evaluated in a single visit protocol at the onset of a perioral outbreak, consistent historically and clinically with recurrent Herpes labialis. Direct immunofluorescence of cytological smears of the lesions/oral fluids was used to confirm Herpes simplex virus types I or II. Patients were randomly assigned to one of two treatment groups: (1) active ingredient and (2) sterile water control. The viral lesion was evaluated as to clinical stage according to standard protocol. Salivary fluid samples were taken: (1) at baseline; (2) immediately following a 30 s rinse; (3) 30 min. after the 30 s rinse; and (4) on the repeat trial, also at 60 min. after the 30 s rinse. All samples were evaluated for viral titer and results compared. RESULTS: In Trial 1, the sample population consisted of 19 males and 21 females with an average age of 29.2 and in Trial 2, 21 males, 19 females with an average age of 28. In both Trials 1 and 2, recoverable infectious virions were reduced to zero after a 30 s experimental rinse; whereas, the control rinse resulted in a non-significant (p>0.05) reduction. The experimental group also demonstrated a continued significant (p<0.05) reduction 30 min. post rinse when compared with baseline while the control group returned to baseline levels. In Trial 2, the 60 min. post rinse follow-up demonstrated a 1-2 log residual reduction from baseline in the experimental group; however, this was not significant. CONCLUSIONS: There is clinical efficacy in utilizing an oral rinse with the antimicrobial agent Listerine Antiseptic in reducing the presence of viral contamination in oral fluids for at least 30 min. after oral rinse. The risk of viral cross contamination generated from these oral fluids in person to person contact or during dental treatment may be reduced. PMID- 15811050 TI - Soluble beta-1,3/1,6-glucan from yeast inhibits experimental periodontal disease in Wistar rats. AB - OBJECTIVE: We have investigated whether a purified immunomodulatory water soluble beta-1,3/1,6-glucan isolated from the cell wall of Bakers yeast, Saccharomyces cerevisiae, would influence the progression of ligature-induced periodontal disease, and to modulate accompanying cytokine and hypothalamic-pituitary-adrenal (HPA) axis responses to a lipopolysaccharide (LPS) challenge. MATERIAL AND METHODS: beta-1,3/1,6-glucan (10 mg/kg/day) was given in the drinking water to Wistar rats during the entire experiment, starting 14 days before disease induction, while control rats were given tap water only. Periodontal disease was assessed when the ligatures had been in place for 35 days. RESULTS: Orally administered soluble beta-1,3/1,6-glucan significantly reduced periodontal bone loss as measured on digital X-rays (p=0,026). Glucan-treated rats also showed a significantly enhanced plasma level of the HPA axis-driven hormone corticosterone (p=0.047), and of the cytokine transforming growth factor-1beta (p=0.032), as well as a tendency to enhanced IL-10 (p=0.106), induced by intra-peritoneally administered LPS. CONCLUSION: Soluble beta-1,3/1,6-glucan administered by the oral route diminishes ligature-induced periodontal bone loss in this model. This effect may be attributable to the well documented ability of beta-1,3/1,6-glucan to stimulate macrophage phagocytosis and to skew the T helper (Th)1/Th2 balance towards Th1 and T regulatory responses. The HPA axis may play a significant role in beta-1,3/1,6-glucan induced immune modulation. PMID- 15811051 TI - Areca nut extracts modulated expression of alkaline phosphatase and receptor activator of nuclear factor kappaB ligand in osteoblasts. AB - OBJECTIVES: Areca chewers have a higher prevalence of periodontal diseases than non-chewers. This study was to determine the possible effects of ripe areca nut extracts (rANE) on viability and gene expression of alkaline phosphatase (ALP), receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in human osteoblasts. METHODS: The effects of rANE on cell viability of osteoblast-like MG63 cells were determined using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) that measures metabolic activity. Gene expression of ALP, RANKL and OPG was examined using reverse transcription polymerase chain reaction. ALP activity and RANKL protein were further examined using substrate assay and confocal laser scanning microscopy, respectively. RESULTS: Relative viability was reduced to approximately 50% when 25 microg/ml of rANE was used. The expression of OPG mRNA in MG63 cells was not altered by rANE. However, decreased levels of mRNA and enzyme activity of ALP were observed. Moreover, the expressions of mRNA and protein of RANKL were stimulated by rANE in a dose-dependent manner. CONCLUSIONS: The rANE affected morphology and viability of osteoblasts. We also present novel evidence demonstrating that areca nut may compromise the periodontal health of areca chewers by suppression of ALP gene expression and elevation of RANKL gene expression in osteoblasts. PMID- 15811052 TI - Efficacy of the Cybersonic in comparison with the Braun 3D Excel and a manual toothbrush. AB - OBJECTIVES: To evaluate the efficacy of two power toothbrushes (Cybersonic and Braun 3D Excel) and one manual brush (Elmex super 29). MATERIAL AND METHODS: After professional toothcleaning 120 subjects were randomly assigned to three groups. Four weeks later, at baseline, the Quigley-Hein plaque index (QHI), the modified approximal plaque index (API), and the papillary bleeding index (PBI) were recorded. Thereafter the subjects used the assigned toothbrushes for 8 weeks. Improvements of the indices after 4 and 8 weeks were calculated as medians. Kruskal-Wallis- and Mann-Whitney U-test served for statistical analysis. RESULTS: All indices showed statistically significant reductions for both power toothbrushes which were superior to the manual brush (4 weeks: Cybersonic: API, 0.21; PBI, 0.25; QHI, 0.23; Braun: API, 0.20; PBI, 0.39; QHI, 0.22; Elmex: API, 0.04; PBI, 0.02; QHI, 0.07; 8 weeks: Cybersonic: API, 0.28; PBI, 0.36; QHI, 0.41; Braun: API, 0.36; PBI, 0.61; QHI, 0.35; Elmex: API, 0.03; PBI, 0.10; QHI, 0.08; p<0.001). The Braun was superior to the Cybersonic with respect to API after 8 weeks (p<0.05) and PBI after 4 and 8 weeks (p<0.01). CONCLUSION: Cybersonic and Braun 3D Excel may be more efficacious than a manual toothbrush in removing plaque and reducing gingivitis. PMID- 15811053 TI - Relationship between attitudes toward oral health at initial office visit and compliance with supportive periodontal treatment. AB - OBJECTIVES: The objectives of the present study include investigation of the relationship between attitudes and desires with respect to oral health at initial office visit and compliance with supportive periodontal treatment (SPT) and identification of prognostic factors with respect to low-compliance with SPT. MATERIALS AND METHODS: Four hundred thirty-one patients were evaluated. Subjects completed a questionnaire concerning attitude and desire with respect to oral health and subjective symptoms prior to periodontal treatment. Survival probabilities of SPT were estimated by the Kaplan-Meier method and compared between answers for each item of the questionnaire via the Cox-Mantel test. Finally, a multivariate Cox proportional hazards regression model was constructed, which included age and gender. RESULTS: Greater than 95% of participants desired toothbrushing proficiency and lifelong retention of teeth at the initial office visit; however, the overall survival probabilities of SPT were only 52.7% after about 5 years. Patients exhibiting unfavourable attitudes toward oral health at the initial office visit, in comparison with those displaying favourable attitudes, exhibited greater tendency to abandon SPT. A Cox regression model revealed that lack of brushing on the gingival margin, non-use of an inter dental brush or dental floss, non-use of fluoride toothpaste and frequent consumption of sugar-containing drinks were significant independent prognostic factors for low-compliance with SPT (p<0.05; Hazard ratios=2.27, 2.00, 2.56 and 2.06, respectively). CONCLUSIONS: Desire for satisfactory oral health is not related consistently to continuation of SPT. Unfavourable attitudes toward oral health were correlated to low-compliance with SPT. Clinicians may wish to establish methods for improvement of patient compliance employing behavioural approaches applicable to the attitudes of potential low-compliance individuals. PMID- 15811054 TI - The potential role of interleukin-17 in the immunopathology of periodontal disease. AB - BACKGROUND: Interleukin-17 (IL-17) is exclusively produced by activated T cells, and this cytokine can induce inflammatory responses, support immune responses (Th1), and stimulate osteoclastic bone resorption in combination with receptor activator of NF-kappaB (RANK) and RANK ligand (RANKL). These biological functions are relevant to the aetiopathogenesis of periodontitis, and thus we sought to investigate whether IL-17 is produced in periodontal lesions and to assess the relationship of gene expression between IL-17 and other cytokines, and to determine the effect of IL-17 on IL-6 production in human gingival fibroblasts (HGF). MATERIALS AND METHODS: IL-17 was detected and measured in periodontal tissues obtained as biopsy samples during periodontal surgery and in the cell free culture supernatants cultured ex vivo, by using Western immunoblotting and enzyme-linked immunosorbent assay, respectively. IL-17 and other cytokine gene expression were investigated by the reverse transcription-polymerase chain reaction (RT-PCR) method. The contribution of IL-17 to IL-6 production by HGF was studied. RESULTS: IL-17 protein was moderately detected in periodontal tissues. In contrast, IL-17 mRNA was expressed only in nine of 23 periodontitis tissue samples by RT-PCR. The IL-17 mRNA-positive samples simultaneously expressed mRNAs encoding interferon (IFN)-gamma, IL-2, RANK, and RANKL, but not IL-4. IL-10 (Th2 cytokine) was detected more frequently in the samples than IFN-gamma and IL-2 (Th1 cytokine). Recombinant human IL-17 induced IL-6 production from HGF in a dose- and time-dependent fashion. CONCLUSIONS: These results indicate that IL-17 is produced in periodontal lesions, which may be involved in Th1 modulation and enhance inflammatory reactions via gingival fibroblast-derived mediators in periodontal disease. Thus, IL-17, together with other cytokines, has a potential role in the aetiopathogenesis of periodontal disease. PMID- 15811055 TI - Interest in periodontology and preferences for treatment of localized gingival recessions. AB - OBJECTIVES: The purpose of this study was to evaluate the views, knowledge and preferences of a large sample of practising dentists in periodontics, focusing specifically on the treatment of gingival recessions, and to compare the findings with the current evidence available in the dental literature. METHODS: We conducted a cross-sectional postal survey of 3780 dentists, representing the majority of all dentists working in Switzerland. The questionnaire consisted of 17 questions, most of them giving the possibility of multiple choices of answers. The demographic profile, interests and satisfaction in periodontics were associated with the choice of treatment options offered for the management of six clinical situations. RESULTS: One thousand two hundred and one dentists sent back the questionnaire within three months and were thus included in the analysis. In general, the interest and the satisfaction in periodontics were moderate to high (6-7 on an analogue scale from 1 to 10). Specialists in periodontics indicated a significantly higher interest and satisfaction in periodontics than the general dentists (p<0.001), and practitioners working in urban areas indicated a slightly higher interest (p=0.027) and satisfaction (0.047) than their colleagues established in a rural setting. The predominant indication of root coverage procedures was aesthetics (90.7%). The region in which dentists worked was the only significant predictor for choosing "no treatment" of buccal recessions: dentists from the German-speaking part were significantly less inclined to surgically treat gingival recessions than their colleagues from the French or Italian part. For those who opted for therapy, a free tissue graft was generally the favourite option, followed by a connective tissue graft and a coronally advanced flap. Throughout, only a small fraction of the dentists considered using a guided tissue regeneration procedure. The relative odds for not extracting teeth with severe periodontal disease were higher if the dentist was a specialist than a generalist. Satisfaction in practicing periodontics also positively strengthened the inclination towards keeping severely compromised teeth. CONCLUSIONS: Aesthetic concerns were the predominant indication for root coverage procedures. Further research should therefore include aesthetic aspects as primary clinical outcome variables. Specific training of dentists and their satisfaction in periodontics influenced treatment decisions. Specialists involved in continuing education should inform practicing dentists more efficiently on the potential and usefulness of periodontal therapy for saving and maintaining periodontally compromised teeth. PMID- 15811056 TI - Levels of interleukin-17 in gingival crevicular fluid and in supernatants of cellular cultures of gingival tissue from patients with chronic periodontitis. AB - BACKGROUND AND AIMS: Interleukin-17 (IL-17) is a T-cell-derived cytokine that may play an important role in the initiation or maintenance of the pro-inflammatory response and has recently been found to stimulate osteoclastic resorption. The purpose of the present study was to determine the presence of IL-17 in gingival crevicular fluid (GCF) samples and in the culture supernatants of gingival cells from patients with chronic periodontitis. METHOD: GCF samples were collected during 30 s from two sites in 16 patients from periodontally affected sites (probing depth > or =5 mm, attachment loss > or =3 mm). The comparison with healthy controls was carried out by collecting GCF samples from eight healthy volunteers. GCF was collected using a paper strip and ELISA was performed to determine the total amount of IL-17. Supernatant cellular cultures of gingival cells were obtained from periodontal biopsies taken from 12 periodontitis patients and from eight healthy control subjects during the surgical removal of wisdom teeth. Spontaneous and phytohaemagglutinin (PHA)-stimulated levels of IL 17 were determined by ELISA. RESULTS: The total amount of cytokine IL-17 was significantly higher in the periodontitis group than the control group (45.9 versus 35.6 pg, p=0.005). Significantly higher GCF volume and amount of total proteins were obtained from periodontitis patients as compared with control subjects (0.98 versus 0.36 microl, p=0.0005; 0.12 versus 0.05 microg, p=0.0005, respectively). A higher concentration of IL-17 was detected in culture supernatants from periodontitis patients compared with healthy subjects, either without stimulation (36.28+/-8.39 versus 28.81+/-1.50 microg/ml, p=0.011) or with PHA stimulation (52.12+/-14.56 versus 39.00+/-4.90 microg/ml, p=0.012). Treatment with PHA induced a significant increase in the production of IL-17 in healthy subjects and periodontitis patients (p=0.001 and 0.003). CONCLUSIONS: The total amount of cytokine IL-17 in GCF samples and in the culture supernatants of gingival cells are significantly increased in periodontal disease. PMID- 15811057 TI - A 0.05% cetyl pyridinium chloride/0.05% chlorhexidine mouth rinse during maintenance phase after initial periodontal therapy. AB - BACKGROUND: Chlorhexidine (CHX) mouth rinse/spray can still be considered the gold standard in the chemical prevention of plaque formation and development of gingivitis. The product unfortunately has some side effects, such as extrinsic tooth staining, poor taste, taste disturbance, sensitivity changes in tongue, pain and irritation because of the alcohol content. These side effects led to the search of new formulations. METHODS: In this double-blind, randomized, long-term, parallel study, 48 moderate periodontitis patients rinsed for 6 months (starting immediately after a "one-stage, full-mouth" disinfection) with one of the following products: CHX 0.2%+alcohol (Corsodyl), CHX 0.05%+ cetyl pyridinium chloride (CPC) 0.05% and no alcohol (Perio-Aid Maintenance, a new formulation), or the placebo of the latter. After 1, 3 and 6 months a series of clinical and microbiological parameters were recorded for the supra- and subgingival area as well as for saliva. RESULTS: Although there was a significant treatment impact (mechanical debridement) in all groups, both CHX solutions further decreased both plaque and gingivitis indices (p<0.001 and p<0.05, respectively), when compared with placebo. This was also reflected by additional reductions in the number of CFU/ml of aerobic and especially anaerobic species and by a suppression of Streptococcus mutans (versus an overgrowth for the placebo), in all niches. Differences between both CHX solutions were never encountered. The subjective ratings were slightly in favour of the new CHX-CPC formulation when compared with the other CHX-alcohol formulation, especially for taste of the product (p<0.05), but less impressive for the staining of teeth and tongue. CONCLUSIONS: The results of this study demonstrated the potential of a new CHX 0.05%+CPC 0.05% non alcoholic formulation as an effective antiplaque agent for long-term use with reduced subjective side effects. PMID- 15811058 TI - Periodontal dressing (Vocopac) influences outcomes in a two-step treatment procedure. AB - OBJECTIVES: It is not clear if periodontal dressing influences the long-term results in a non-surgical treatment procedure. MATERIAL AND METHODS: The periodontal parameters (pre-baseline) of 36 patients with aggressive periodontitis were obtained before the patients were treated initially (1st step) by a dental hygienist, who completely removed the supra- and subgingival concrements. Baseline parameters were raised 3 weeks after the 1st step, before the 2nd therapy step was conducted. It consisted of a non-surgical procedure, which comprised a closed full-mouth manual root curettage (root planing), immediate systemic application of metronidazole, and the placement of a periodontal dressing (Vocopac, Voco). The patients were randomized to two test groups having their periodontal packs removed after 3-4 days (group 1, n=12) and 7-8 days (group 2, n=12), respectively and a control group (n=12) without periodontal dressing. Clinical parameters were raised again after 6 and 24 months. RESULTS: Six and 24 months later, changes in probing pocket depth (PPD) and probing attachment level (PAL) were observed in all three groups compared with baseline, but the difference was significant in group 2 only. In addition, group 2 showed a greater reduction in mean PPD and also a significantly greater gain of attachment in comparison with the controls. CONCLUSION: Wound dressing has a positive effect on clinical long-term results using a two-step non-surgical procedure. Moreover, removing the dressing after 7-8 days leads to clearly better results than removing it earlier. PMID- 15811059 TI - Scaling and root planing, systemic metronidazole and professional plaque removal in the treatment of chronic periodontitis in a Brazilian population II- microbiological results. AB - OBJECTIVE: The current investigation evaluated changes in levels and proportions of 39 bacterial species in subgingival plaque samples after scaling and root planing (SRP) alone or in combination with systemic metronidazole and/or professional cleaning in subjects with chronic periodontitis. METHODS: Forty-four adult subjects (mean age 45+/-6 years) with periodontitis were randomly assigned in four treatment groups, a control (C, n=10) that received SRP and placebo and three test groups treated as follows: T1 (n=12): SRP and metronidazole (M, 400 mg tid) for 10 days; T2 (n=12): SRP, weekly professional supragingival plaque removal for 3 months (PC) and placebo; and T3 (n=10): SRP, M and PC. Subgingival plaque samples were taken from seven sites per subject at baseline and 90 days post-therapy. Counts of 39 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Wilcoxon signed ranks test and among groups using ancova. RESULTS: The mean counts of the majority of the species were reduced post-therapy in the 4 treatment groups. Counts (x 10(5)+/-SEM) of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola were significantly reduced in groups T2 and T3. Levels of beneficial species, such as some Actinomyces species, Veillonella parvula, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii were minimally affected in levels when the combined therapy was applied (T3). Mean proportions of red complex species decreased from 18.4% at baseline to 3% at 90 days post-therapy in group T3 (p<0.01), from 25.8% to 2.3% in group T2 (p<0.01), from 17.7% to 5.6% in group T1 (p<0.05) and from 19.4% to 8.8% in group C (NS). Proportions of the suspected periodontal pathogens from the orange complex were also markedly reduced in groups T2 and T3. CONCLUSIONS: All treatments reduced counts and proportions of red complex species. Adjunctive therapy appeared to have a greater effect and also affected members of the orange complex. PMID- 15811060 TI - Periodontal infection as a possible severity factor for rheumatoid arthritis. AB - OBJECTIVE: Clinical effects of periodontal treatment on biochemical and clinical markers of disease severity in rheumatoid arthritis (RA) patients with periodontal disease were evaluated. METHODS: Forty-two patients were assigned to two groups, G1 (n=16) and G2 (n=26). G1 patients were submitted to oral hygiene instruction and professional tooth cleaning and G2 patients additionally had full mouth scaling and root planing (SRP). Clinical periodontal measurements were obtained at baseline and 3 months after periodontal treatment. A Health Assessment Questionnaire (HAQ) was used to evaluate their performance on daily living. Rheumatoid factor (RF), erythrocyte sedimentation rate (ESR) and drug therapy were assessed. RESULTS: Both groups presented a full-mouth improvement in all periodontal clinical parameters (p<0.05), with the exception of clinical attachment level (CAL) and probing pocket depth (PPD) >6 mm for G1. G2 showed greater mean reductions on PPD >4 mm than G1 (p<0.001). HAQ analyses showed a reduction on the degree of disability of G2, but not statistically significant. ESR was significantly reduced for G2 after SRP although RF did not show statistical reductions. CONCLUSION: The data suggest that periodontal treatment with SRP might have an effect on the ESR reduction. PMID- 15811061 TI - Plasminogen activator system in smokers and non-smokers with and without periodontal disease. AB - BACKGROUND: The present study assessed levels of plasminogen activator (PA) system proteins in gingival crevicular fluid (GCF) and serum of chronic gingivitis, chronic periodontitis patients and periodontally healthy subjects and evaluated how smoking influenced these levels. METHODS: Twenty chronic gingivitis; 20 chronic periodontitis patients and 20 periodontally healthy volunteers were consecutively recruited according to the inclusion criteria so that exactly half of the subjects in each category were smokers. GCF samples from four sites together with serum samples were obtained from each subject. GCF levels of tissue type PA (t-PA), urokinase type PA (u-PA), PA inhibitor-1 (PAI-1) and PA inhibitor-2 (PAI-2) and serum concentrations of cotinine, u-PA and PAI-1 were analysed by enzyme-linked immunosorbent assay. RESULTS: The only statistically significant difference between smokers and non-smokers was a lower GCF PAI-2 concentrations in healthy smokers compared with healthy non-smokers (p<0.01). Gingivitis and periodontitis patients had higher GCF concentrations of PAI-2 than healthy subjects (p<0.002 and p<0.02 respectively). The ratio of u PA:PAI-1 and t-PA:PAI-1 were significantly higher in GCF of smokers with periodontitis compared with "healthy" smokers, whereas the ratio of t-PA:PAI-2 was significantly lower in smokers with periodontal disease (p<0.05). CONCLUSIONS: GCF levels of the PA system proteins are increased in chronic gingivitis and periodontitis compared with healthy gingiva. Smoking had only subtle effects on the GCF PA system proteins with the exception of PAI-2, and the balance of activators and inhibitors. These findings suggest one mechanism whereby smoking may exert detrimental effects on the periodontal tissues. PMID- 15811062 TI - A new ultrasonic device in maintenance therapy: perception of pain and clinical efficacy. AB - BACKGROUND: A recently introduced piezo-driven ultrasonic device (Vector) generates longitudinal oscillations. As a result, the instrument tip moves parallel to the tooth surface. By avoiding vertical oscillations, maintenance treatment with the Vector device should be less painful than treatment with conventional systems. We investigated whether patients perceive treatment with the Vector device as less painful than with a conventional ultrasonic device, and whether the clinical efficacy of the Vector device is comparable with that of the conventional ultrasonic device in maintenance patients. MATERIAL AND METHODS: Thirty-eight maintenance patients with moderate to advanced periodontal disease took part in this prospective, randomized controlled clinical study. Each patients had to have at least two teeth with probing depths of >4 mm. They were treated either with Dentsply (n=22) at a reduced power setting or with the Vector device (n=16). The observation period was 6 months. Probing pocket depth, attachment level, and bleeding upon probing were assessed at six sites on each treated tooth by a blinded investigator Patient were asked to report perceived pain during instrumentation with a visual analog scale immediately after treatment, in the evening of the treatment day, and in the evenings 1 and 2 days after treatment. RESULTS: Bleeding on probing, probing depth, and attachment level improved in both instrumentation groups from baseline to month 6; however, there was no difference between the two instrumentation modalities. The patients perceived treatment with neither instrument as unpleasant, and their perception of pain intensity both during instrumentation and on the following days did not differ. CONCLUSION: In maintenance therapy, clinical efficacy of the vector device is comparable with that of conventional ultrasonic device. It makes no difference whether the ultrasonic device at a reduced power setting or the Vector device is used, since patients perceive both instruments as causing very little pain. PMID- 15811063 TI - Gingivo-mucosal and cutaneous reactions to amalgam fillings. AB - BACKGROUND: A number of reports exist on the side effects of materials used to restore teeth. Most of the cases involve local allergy reactions, but also skin lesions are described. Few cases are described where both local effects on the mucosa and skin lesions distant to the oral cavity are caused by amalgam. RESULT: The case presented indicates that the release of mercury from amalgam fillings is able to induce hypersensitivity reactions resulting in soft-tissue changes in the gingiva, buccal mucosa, tongue and on the skin of the back of the hands. PMID- 15811065 TI - Clinical comparison of the "typical oscillating/rotating powered toothbrush" and the "conventional manual toothbrush". PMID- 15811066 TI - The impact of mitochondrial genetics on male infertility. AB - Summary Human mitochondrial DNA (mtDNA) encodes 13 of the polypeptides associated with the process of oxidative phosphorylation (OXPHOS), the cells most important ATP generating pathway. Until recently, the effects of mtDNA rearrangements on male fertility have been largely ignored. However, it is becoming increasingly evident that both point mutations and large-scale deletions may have an impact on sperm motility and morphology. We discuss the implications of these rearrangements in the context of the clinical setting. We further discuss the possible consequences resulting from the transmission of sperm mtDNA deletions to the offspring. The role of nucleo-cytoplasmic interaction is investigated in the context of nuclear transcription and replication factors that regulate mtDNA transcription and replication. PMID- 15811067 TI - A neglected gland: a review of Cowper's gland. AB - Cowper's glands are pea sized glands present inferior to the prostate gland in the male reproductive system. They produce thick clear mucus prior to ejaculation that drains into the spongy urethra. Though it is well established that the function of the Cowper's gland secretions is to neutralize traces of acidic urine in the urethra, knowledge regarding the various lesions and associated complications of this gland is scarce. This review provides a comprehensive report on the development, function and various lesions associated with Cowper's gland. PMID- 15811068 TI - The physiological basis of human sexual arousal: neuroendocrine sexual asymmetry. AB - Normal sexual arousal and response suppose an integrated process involving both physiological and psychological processes. However, the current understanding of sexual arousal does not provide a coherent model that accounts for the integration of multiple physiological systems that subsequently generate a coordinated sexual response at both the spinal peripheral and cerebral central levels. Herein we suggest a model that involves both sympathetic and parasympathetic activation during sexual arousal via the two classes of gonadal hormones, androgens and oestrogens. We discuss the manner in which gonadal hormones may activate such a system, transforming pre-pubertal (non-erotic) genital stimulation to post-pubertal erogenization of stimulation and subsequent sexual arousal. Finally, we indicate that the different balance of androgens and oestrogens in men and women may generate asymmetric effects on each of the components of the autonomic nervous system, thereby explaining some of the differences in patterns of sexual arousal and the responses cycle across the sexes. PMID- 15811069 TI - Penile veins play a pivotal role in erection: the haemodynamic evidence. AB - Although penile venous surgery has almost been abandoned and the venous factor eliminated as a contributing factor to erectile dysfunction, new concepts of erection-related veins has recently been described and reported in literature. We sought to conduct a haemodynamic study on human cadavers in order to elucidate to what extent penile veins act in erection, and to explore the possible role of erection-related veins as an important contributor to impotence. From November 2002 to December 2003, seven fresh human cadavers of men who had no sexual activity for at least 6 months prior to death, and in whom the penis was intact were used for this study. Infusion cavernosometry was carried out with an induction flow of 150 mL/min before and after the erection-related veins were removed. A rigid erection was attained in all subjects, lasting significantly longer (p = 0.043) after removal of erection-related veins. Similarly, there were significant differences in the maintenance flow (p = 0.043), T(max) (p = 0.043), V(max) (p = 0.043), and pressure loss (p = 0.043). In cadaveric penises, a rigid erection could be maintained in spite of the fact that the low flow rate of 21 mL/min is much lower than the average arterial perfusion rate observed in cases of arterial insufficiency. We therefore concluded that penile veins may play a significant role in attaining sufficient erection, and further research is required to study this possible clinical implication. PMID- 15811070 TI - Improvement of semen quality by nocturnal scrotal cooling in oligozoospermic men with a history of testicular maldescent. AB - For infertile men with a history of testicular maldescent only few therapeutic options exist beside assisted reproduction. The aim of our study was to evaluate the influence of nocturnal scrotal cooling on semen quality in such patients presenting with oligozoospermia. Twenty infertile men with a history of testicular maldescent and oligozoospermia were included for nocturnal scrotal cooling over 12 weeks for every night. To increase nocturnal periscrotal air circulation we used a membrane pump connected via plastic tubes to receptacles placed in both groins. Semen analysis was performed at the beginning of the cooling period and at weeks 4, 8 and 12. Another 20 infertile patients with a history of testicular maldescent and oligozoospermia were followed without specific treatment and served as a retrospectively built control group. Scrotal cooling at night by means of a perigenital air stream resulted in a scrotal temperature drop by 0.8 degrees C (median). A significant increase in sperm concentration and total sperm count was achieved by nocturnal cooling after 8 weeks (p < 0.01; p < 0.05; respectively) and 12 weeks (p < 0.01; p < 0.01; respectively). The improvement of sperm motility and sperm morphology was statistically insignificant. The present study suggests nocturnal scrotal cooling as a therapeutic option to improve semen quality. In a further controlled prospective study the influence on pregnancy rates should be evaluated. PMID- 15811071 TI - Ontogenetic profile and thyroid hormone regulation of type-1 and type-8 glucose transporters in rat Sertoli cells. AB - The glucose transporters (GLUTs) gene encode glycoproteins responsible for facilitating transfer of glucose across plasma membrane. In testis, different members of this family are present. In particular the main GLUT mRNA expression within the adult testis is the type 8, while type 1 is more expressed in prepubertal testis. Thyroid hormone, which receptors and function have been characterized in the testis, plays a crucial role in the cellular energetic metabolism. In fact, in the immature Sertoli cells, GLUT1 is up regulated by l triiodothyronine (T(3)). The aim of this paper is to investigate the expression profile of GLUT1 and GLUT8 in the testis during development and in adulthood and analyse the role of T(3) on their expression. To analyse the expression of GLUT8 and GLUT1 we performed Northern blot and RT-PCR experiments in the whole testis and in Sertoli cells from rats of different ages. Treatments in vivo and in vitro with T(3) were used to study the effect of thyroid hormones on GLUT1 and GLUT8 expression. The activity of the rat GLUT1 promoter and its regulation by T(3) was studied with transient transfections in gonadal and non-gonadal cell lines and in primary Sertoli cell cultures. GLUT8 is expressed at a low level in the prepubertal testis and Sertoli cells and does not appear to be under T(3) control. GLUT1 is the predominant form in immature Sertoli cells. The effect of T(3) on its mRNA accumulation was quantified and confirmed by RT-PCR (control: 0.65 +/- 0.17; T(3): 1.23 +/- 0.04, arbitrary units, p < 0.05). However, transfection experiments showed that T(3) does not directly regulate GLUT1 promoter in any cell line tested. This is confirmed by the evidence that, upon extensive analysis, the rat GLUT1 promoter and the first intron sequence do not shows any thyroid responsive elements. Our data demonstrate that GLUT1 and GLUT8 are both expressed in prepubertal testis, but only GLUT1 is regulated by T(3). In addition, we found that the effect of T(3) cannot be attributed to its action on GLUT1 promoter. PMID- 15811072 TI - A new and simple method to evaluate early membrane changes in frozen-thawed boar spermatozoa. AB - Detection of early changes in the sperm plasma membrane during cryopreservation is of utmost importance when designing freezing protocols and has previously been studied in the pig species using annexin-V detection of phosphatidylserine translocation. In the present study we designed a new assay to detect these changes in boar spermatozoa, based on the slight increase of sperm membrane permeability occurring during the early stages of cryoinjury, using the combination of three fluorescent probes, SNARF-1, YO-PRO-1 and ethidium homodimer. Four ejaculates from five different boars were frozen-thawed and flow cytometrically (FC) evaluated as paired samples. One of the samples was assayed using the annexin-V/propidium iodide staining and the other sample was evaluated using the new triple staining. Using this combination of probes, four sperm subpopulations were easily detected: viable, with stable membranes (SNARF-1 positive cells), and three with compromised membranes, one of YO-PRO-1+/Eth- cells, one ethidium homodimer+ spermatozoa and, finally spermatozoa stained both with YO-PRO-1 and ethidium homodimer (YO-PRO-1+/Eth+). The latter three categories corresponded to dead spermatozoa, but with different degree of membrane damage, being YO-PRO+/Eth- an earlier stage of membrane destabilization, (manifested by an increase in membrane permeability, while still maintaining membrane integrity) than YO-PRO+/Eth+. A method agreement analysis between both methods was performed revealing good agreement, although the percentage of live cells was 9.44% larger for the triple stain than the annexin-V assay. The new assay stained all sperm sub-populations present in the sample, making it especially suitable for both fluorescence microscopy and flow cytometry, facilitating the exclusion of debris and egg-yolk particles when using FC. PMID- 15811073 TI - Mutation C677T in the methylenetetrahydrofolate reductase gene is associated with male infertility in an Indian population. AB - A mutation (C677T) in the gene, MTHFR, is known to increase susceptibility to various multifactorial disorders. In order to assess this single nucleotide polymorphism (SNP) as risk factor for idiopathic male infertility, a case-control study was done on an Indian population. DNA from 151 cases of non-obstruction, idiopathic oligo-/azoospermia and 200 fertile males (controls) was polymerase chain reaction amplified using site-specific primers, and analysed for the mutation following HinfI-digestion. Our results show a significantly increased frequency of CT heterozygotes among infertile patients (p value <0.04). More importantly, while there were no T homozygotes in the control population, six of 151 infertile cases were T homozygous. Considering that T allele occurs in very low frequency in the control population, 677T is clearly a risk factor for infertility in the Indian population. We contend that the same could also be true for African and Southeast Asian populations where the frequency of 677T is very low. The lack of similar association in western populations could be because of the overall dietary enrichment of folates, which could nullify or minimize the effect of this polymorphism. PMID- 15811074 TI - Characterization of a critical region in the hormone binding domain of sperm progesterone receptor. AB - Our previous studies have demonstrated that the ligand binding characteristics of the non-genomic sperm progesterone receptor (sPR) are different from those of the conventional progesterone receptor (PR). Unlike PR, sPR does not bind efficiently to progesterone antagonists i.e. mifepristone (RU38486), onapristone (ZK98.299) etc. The present study was undertaken to determine the molecular composition of a region that plays a critical role in the interaction of the hormone-binding domain (HBD) of sPR with progesterone and its antagonists. Detection, cloning and sequencing of the HBD region of sPR revealed its complete sequence homology to the corresponding region in the conventional PR. No nucleotide substitution/mutation/deletion, which could account for the differential antihormone binding, was observed in the HBD of sPR. The critical codon (nucleotide 3216-3218) in all three clones for the HBD of sPR was found to encode for glycine. This ruled out the possibility of steric hindrance because of the placement of amino acids with side chains at a critical position in the HBD, which may interfere in binding of sPR with antiprogestins. It is likely that the post-transcriptional modifications contribute to the differential binding characteristics of sPR. This warrants future investigations to focus more on the characterization of the mature sPR protein. PMID- 15811075 TI - Willan's itch and other causes of pruritus in the elderly. AB - Itch in the elderly presents a diagnostic and therapeutic challenge. A thorough history, review of systems, and physical examination are critical to determining its cause. Examination of the skin may be misleading. There are frequently only secondary lesions, eczematous changes, lichenification, and excoriation, which may be misdiagnosed as a primary dermatitis. Xerosis may be the cause, but it is sometimes merely coincidental. If primary lesions are present, a skin biopsy can enable a diagnosis to be made. Systemic causes of itch, such as cholestasis, uremia, hyperthyroidism, medications, or lymphoma, must be considered. If the cause remains elusive, idiopathic itching of the elderly or so-called "senile pruritus" may be considered. However, we propose to discard the term "senile pruritus", which can be offensive and frightening. We propose to replace it with "Willan's itch". Robert Willan (1757-1812) is honored as one of the founders of modern dermatology thanks to his book, On Cutaneous Diseases, and its morphological approach to skin disease. He was probably the first to give a good clinical description of itching in the elderly. The diagnosis of Willan's itch should be reserved for generalized pruritus in the absence of xerosis or other recognizable cause. The pathophysiology of this form of pruritus is poorly understood, but it is likely that age-related changes of the skin, cutaneous nerves, and other parts of the nervous system play a role. Anecdotal and limited data suggest that gabapentin, cutaneous field stimulation, serotonin antagonists, and ultraviolet B phototherapy may attenuate itch in some of these patients. PMID- 15811076 TI - Clinical, histopathological and virological findings in patients with focal epithelial hyperplasia from Colombia. AB - BACKGROUND: Focal epithelial hyperplasia (FEH) is a rare oral HPV-related disease, highly prevalent in certain ethnic communities. A previous study found 7.5% prevalence among school children from the Indian community Embera-Chami in Colombia. OBJECTIVE: To determine the prevalence and Human Papillomavirus (HPVs) type in children with FEH. MATERIALS AND METHODS: One hundred and thirty-eight students were screened. All FEH samples were HPV-tested by two PCR-based systems. RESULTS: We identified 18 FEH cases. beta-globin amplification was obtained in 15 cases and nine were HPV-55-positive by the HPV-PCR-hybridization method. Nine cases (50%) were HPV-13-positive by the GP5+/GP6+-based method. Twelve cases (80%) were HPV-positive by one or the other method. Forty-four percent and 88% of interviewed parents reported family histories of FEH and toothbrush sharing, respectively. CONCLUSIONS: Human Papillomavirus-DNA was identified in the majority of FEH cases and HPV-13 was the only genotype involved. Frequent familiar infection suggests interfamiliar transmission, genetic predisposition or infection susceptibility among family members. PMID- 15811077 TI - Localization of haptoglobin in normal human skin and some skin diseases. AB - BACKGROUND: Haptoglobin (Hp) is an acute-phase reactant, known to be produced mainly in the liver. Haptoglobin can also be detected in the cytoplasm of normal epidermal Langerhans cells (LCs), and can prevent their functional maturation. The synthesis of Hp in skin cells has not been well studied. METHODS: We examined Hp expression at mRNA and protein levels by in situ hybridization and immunohistochemistry, respectively, in normal human skin and in the skin of patients with psoriasis, lichen planus, erythroderma, seborrheic keratosis, verruca vulgaris, basal cell carcinoma, systemic lupus erythematosus, pemphigus and bullous pemphigoid. RESULTS: (1) Haptoglobin mRNA was expressed in the epidermal keratinocytes (KCs), the epithelial cells of hair follicles, sebaceous glands and eccrine glands in normal skin and all dermatoses investigated. (2) Whereas compared with normal skin, the Hp mRNA in KCs of patients with psoriasis, lichen planus, erythroderma, seborrhoea keratosis and verruca vulgaris was significantly intensified, it was weaker in patients with systemic lupus erythematosus, pemphigus and bullous pemphigoid. (3) Haptoglobin protein only stained positively in some KCs of patients with psoriasis, lichen planus and erythroderma. (4) Although some but not all epidermal LCs were positively stained with anti-Hp antibody in normal skin and in skin samples from all patients, the ratios of Hp-positive LCs/total LCs were significantly higher in those diseases with intensified Hp mRNA in KCs. CONCLUSIONS: Skin is another extrahepatic organ where Hp can be synthesized by KCs. The expression of Hp mRNA in KCs and the Hp protein in both LCs and KCs appears to be correlated with the amount of inflammation, which might indicate that skin itself is involved in down regulating the local inflammatory reaction by KC-synthesized Hp. PMID- 15811078 TI - Cardiovascular effects of corticosteroid pulse therapy: a prospective controlled study on pemphigus patients. PMID- 15811079 TI - Complications of the smallpox vaccine. PMID- 15811080 TI - The profile of fogo selvagem (endemic pemphigus foliaceus) at the University Hospital of Brasilia-Brazil. Epidemiological and clinical considerations. AB - BACKGROUND: Endemic pemphigus foliaceus (EPF) or fogo selvagem (FS) is an endemic autoimmune disease, characterized by flaccid bullae induced by IgG(4) subclass antibodies. The authors demonstrate the epidemiological and clinical status of patients who have been followed at the University Hospital of Brasilia (HUB) for more than 15 years. METHODS: One hundred and ninety-six patients with FS took part in the project. In setting up this study a historical descriptive cohort of patients was put together. In order to collect data, the authors used a questionnaire where the patient indicated the sex, age at the onset, occupation, origin, clinical status, including scalp compromise, evolution, cofactors influencing clinical worsening and the treatment compliance. In order to minimize loss to follow up, the authors used the statistical method of incidence density (patients/years). RESULTS: The disease occurred in 58.4% of the young patients in the 11-30-year age bracket, and 52% came from urban areas. These patients included students and teachers. Localized disease predominated as compared with the generalized presentation of this condition. Fifty-nine percent of the patients evolved to the recurrent form. Those patients with the evolutional form, in remission for more than 1 year (94%), had been followed for more than 5 years. Even patients with the less active forms of the disease had scalp lesions. Thus scalp lesions are not an indicator of bad prognosis. CONCLUSIONS: In the present study, the disease affected patients from a higher sociocultural class than previously described. Furthermore, in contrast to other reports, a substantial number of the patients lived in urban areas, although often spending some time in a rural setting for leisure or professional activities. This study suggests that the longer the follow up, the higher the likelihood that the disease would progress to a more controlled clinical presentation. Scalp lesions were not related to adverse prognosis. Sun, heat and infections act as triggering factors for the immunological imbalance, worsening the disease. PMID- 15811081 TI - Lupus vulgaris: report of two cases. AB - Although there has been a steady decline in the incidence of tuberculosis in recent years, it persists in some regions, and where AIDS is especially prevalent, the number of new cases has been increasing. Thus, cutaneous tuberculosis has re-emerged in areas with a high incidence of HIV infection and multidrug-resistant pulmonary tuberculosis. Lupus vulgaris has been and remains the most common form of cutaneous tuberculosis. Cutaneous manifestations of disseminated tuberculosis are unusual, being seen in less than 0.5% of cases. Scrofuloderma, tuberculosis verrucosa cutis and lupus vulgaris comprise most cutaneous tuberculosis cases. Bacillus Calmette-Guerin (BCG) is derived from an attenuated strain of Mycobacterium bovis and is employed beneficially as a relatively safe vaccination in Poland and other countries in which the prevalence of tuberculosis is high. However, BCG vaccination may produce complications, including disseminated BCG and lupus vulgaris, the latter seen in one of our two patients in whom lupus vulgaris at the inoculation site followed a second vaccination with BCG 12 years after the initial one. A similar phenomenon has been described after immunotherapy with BCG vaccination. Re-infection (secondary) inoculation cutaneous tuberculosis may also occur as a result of BCG vaccination, producing either lupus vulgaris or tuberculosis verrucosa cutis, probably depending upon the patient's degree of cell-mediated immunity. However, most lupus vulgaris cases are not associated with vaccination with BCG, as occurred in our first patient. For those who do develop lupus vulgaris, it can be persistent for a long period, in some cases for many decades. In the second patient we describe a lengthy duration and cutaneous reactivation at distant sites after more than 40 years. PMID- 15811082 TI - Nosocomial nasal myiasis owing to Cochliomyia hominivorax: a case in French Guiana. AB - Abstract The larval stages of the fly Cochliomyia hominivorax are responsible for myiasis, which primarily affects wounds. We report the case of a bed-ridden patient with dementia who developed right nasal myiasis during his stay at Cayenne Hospital. Progression was favorable, but the nasal pyramid was partially destroyed. In zones where this fly is endemic, particular attention should be given to hospitalized patients with wounds and consciousness problems. PMID- 15811083 TI - Dermatology in the military: an East Timor study. PMID- 15811084 TI - Localized pemphigoid on the soles of both feet. AB - An 80-year-old woman was referred to our hospital with a spontaneously appearing bullous dermatosis limited to the soles of both feet, causing an intense pruritus. She also suffered from Parkinson's disease and depression, and had been treated with levodopa, benserazide, and mirtazapine for several years. On clinical examination, we found several tense and hemorrhagic bullae, with a diameter of up to 3 cm, at both plantar sites and multiple, confluent, dyshidrotic vesicles ( Figs 1 and 2). The rest of the skin, including the mucous membranes and palms, was normal. The first clinical diagnosis was podopompholyx, but histopathologic findings and direct immunofluorescence revealed a diagnosis of bullous pemphigoid, showing subepidermal blisters (Fig. 3) and linear deposits of C3 and immunoglobulin G (IgG). Indirect immunofluorescence was positive, the IgG autoantibodies bound to the epidermal site of salt-split skin, and circulating antibodies against bullous pemphigoid antigens 1 and 2 were found. Because of this typical clinical picture, a diagnosis of dyshidrotic pemphigoid, a localized form of bullous pemphigoid, was made. Under systemic treatment with prednisolone, 40 mg/day, the skin healed completely within 2 weeks. Descriptions of dyshidrotic pemphigoid limited to the soles of both feet are very rare, and the clinical findings might easily lead to a misdiagnosis of podopompholyx. PMID- 15811085 TI - Familial cutaneous collagenoma: report of an affected family. PMID- 15811086 TI - Lichen sclerosus mistaken for child sexual abuse. PMID- 15811087 TI - Frontal fibrosing alopecia developing after hair transplantation for androgenetic alopecia. PMID- 15811088 TI - Hypertrophic and keloidal scars: an approach to polytherapy. AB - BACKGROUND: Cryotherapy and intralesional corticosteroids have been used separately or in combination as a treatment for hypertrophic and keloidal scars. The addition of silicone gel to the treatment schedule might be of interest. METHODS: Twenty patients with hypertrophic and keloidal scars received two 15-s cycles (in total 30 s) of cryotherapy treatments at every session (once monthly for 12 months) with intralesional injections of trimcinolone acetonide 0.1% (10 40 mg/mL, once monthly for 3 months). Topical application of silicone gel was added three times daily (for 12 months). The control group included 10 patients who received treatment with silicone sheeting. RESULTS: After 1 year there was improvement in all the parameters studied, especially in terms of symptoms, cosmetic appearance and associated signs (P < 0.0001) compared to baseline and compared to the control group. CONCLUSION: Our study suggests that polytherapy may be an effective tool in the therapy of hypertrophic and keloidal scars. PMID- 15811090 TI - Sudden onset of Multiple nevi after administration of 6-mercaptopurine in an adult with Crohn's disease: a case report. AB - The correlation between combined chemotherapy, including 6-mercaptopurine, and development of excessive number of melanocytic nevi in childhood has been previously reported and studied. As of yet, no such relation has been reported in adults. We recently observed a 27-year-old female patient who developed numerous benign melanocytic nevi during a 20-month period while taking 6-mercaptopurine for Crohn's disease. The appearance of many nevi was worrisome to the patient from a cosmetic and medical perspective (given the higher risk of melanoma associated with large numbers of nevi). PMID- 15811089 TI - Pilot trial: Pioglitazone versus placebo in patients with plaque psoriasis (the P6). AB - BACKGROUND: Disordered differentiation and hyperproliferation of keratinocytes with inflammation are the hallmarks of psoriasis. Ligand activation of peroxisome proliferator receptor-gamma (a class of nuclear receptors) by thiazolidinediones can normalize the histologic features of psoriasis. METHOD: In a 10-week, double blind, randomized, placebo-controlled, parallel-group study, 70 patients with moderate to severe psoriasis received one of the following treatments: pioglitazone 15 mg, pioglitazone 30 mg or placebo. Efficacy was evaluated by observing the change in the psoriasis area and severity index (PASI) after 10 weeks of treatment. RESULTS: There was a dose-dependent improvement in psoriasis. Median PASI scores at the end of 10 weeks were significantly reduced in the pioglitazone treatment groups as compared to the placebo-treated group. The psoriasis lesions cleared in more than 40% of patients treated with pioglitazone as compared to 12.5% of those with placebo. The percentage reduction in mean PASI scores was 21.6%, 41.1% and 47.5% in the placebo, pioglitazone 15 mg, and 30 mg groups, respectively. No serious adverse events were detected. CONCLUSION: This is the first report from a controlled trial demonstrating that pioglitazone could be considered as an efficacious and safe agent for the treatment of plaque psoriasis. The optimum dose and duration of pioglitazone therapy remain to be determined. PMID- 15811091 TI - Evaluation of time-dependent response to psoralen plus UVA (PUVA) treatment with topical 8-methoxypsoralen (8-MOP) gel in palmoplantar dermatoses. AB - BACKGROUND: Topical psoralen plus UVA (PUVA) is an effective treatment for localized forms of eczema, psoriasis, and palmoplantar pustulosis, which avoids some of the undesirable side-effects of systemic psoralens. Aims In this study, the efficacy of topical PUVA treatment with 8-methoxypsoralen (8-MOP) gel was compared with placebo plus UVA in chronic recurrent palmoplantar dermatoses. METHODS: Twenty-two patients with palmoplantar disease (11 with psoriasis vulgaris, six with eczema, and five with pustulosis) were enrolled in the study. The study design was a left-right comparison: one hand or foot was treated with 8 MOP 0.01% gel plus UVA, whilst the contralateral hand or foot received placebo and UVA for 6 weeks. Twenty minutes after application of the gel, both sides were exposed to UVA. The treatment regimen was three times a week, and the UVA dose was increased weekly by 20%. RESULTS: A comparison of the pre- and post-treatment scores with regard to the severity of the clinical picture and the infiltration of plaques showed a significant decrease (from 7.5 +/- 2.0 to 2.5 +/- 2.1 and from 2.0 +/- 0.7 to 0.3 +/- 0.5, respectively) in the sites treated with 8-MOP gel compared with placebo after 6 weeks. CONCLUSION: The results of the study indicate that at least 18 courses of local PUVA within 6 weeks, with a cumulative dose of 87 J/cm(2), are required to induce a significant decrease in the disease severity and an improvement in the infiltration of plaques due to 8-MOP gel at a concentration of 0.01% when treating chronic recurrent palmoplantar dermatoses. PMID- 15811092 TI - The mole theory: primary function of melanocytes and melanin may be antimicrobial defense and immunomodulation (not solar protection). PMID- 15811093 TI - Successful treatment of chronic actinic dermatitis with topical pimecrolimus. PMID- 15811095 TI - Changing geography of cutaneous leishmaniasis in the Indian subcontinent and in neighboring Arab states. PMID- 15811096 TI - Case of Mondor's disease of the abdomen (body-building induced). PMID- 15811097 TI - Pseudomonas fluorescens cutaneous abscess and recurrent bacteremia following a dog bite. PMID- 15811098 TI - An outbreak of urticaria among soldiers deploying for combat. PMID- 15811099 TI - Fixed drug eruption and fixed drug-like eruption. PMID- 15811100 TI - HLA-DRB1*0101 is associated with foliaceous pemphigus in Mexicans. PMID- 15811101 TI - Building research by community. PMID- 15811102 TI - Evaluating research for use in practice: what criteria do specialist nurses use? AB - AIM: This paper reports a study identifying the criteria chosen by nurses to evaluate whether to use research in practice. This work is part of a larger project on the process of research use by nurses. BACKGROUND: Prescriptive models of research utilization direct practitioners to evaluate a range of criteria to assess the fit of the research to current practice. Criteria relating to the research, task and nursing context are specified, but it is not known whether nurses use these criteria in practice. METHOD: Three different groups of nurse specialists from a number of health care providers in the North West of England were studied. The specialists were involved in a series of meetings at a local university to construct evidence-based policy recommendations for practice. The discussion was analysed to identify the evaluation criteria they used to assess what should be done in the practical setting. FINDINGS: Three sets of evaluative criteria were identified relating to the research, the task and the fit of the task with the nursing context, and included relevance and quality of the research, effectiveness, practicality, effort and the impact of the task on client and staff, feasibility and fit with the status quo, and the availability of nursing control and feedback from practice. Criteria were domain-, task- and context-specific. CONCLUSIONS: Three frames of reference for decisions relating to the use of research are identified: the debate between efficiency and effectiveness in health care, the difficulties of risk management and the responsibility for risk, and research use as a strategy for control in contested arenas. These frames of reference reflect how nurses are judged in a political and organizational context. Attempts to influence research uptake need to take into account how research is perceived and evaluated by practitioners and how it serves their purposes. PMID- 15811103 TI - Nurse-led models of chemotherapy care: mixed economy or nurse-doctor substitution? AB - AIM: This paper reports a study exploring the perspectives of people affected by cancer (service users) and health care professionals' about current medical consultant-led services and the acceptability of a proposed nurse-led ambulatory chemotherapy service. BACKGROUND: A number of studies have evaluated a nurse-led model of cancer care delivery but little work has been undertaken in chemotherapy settings. Furthermore, many of these studies give little information on how the perspective of users was incorporated in the design and evaluation of these services. METHODS: Service users (n = 26) and health care professionals (n = 22) were recruited across the South East of England. A qualitative study was undertaken... Using a semi-structured interview schedule, participants were asked to give their perceptions of current chemotherapy services and the potential of a nurse-led service. A thematic analysis of data was undertaken. FINDINGS: This paper focuses on the theme of the current and future context of a chemotherapy service. Three sub-themes were identified: contextualizing roles, defining therapeutic outcomes, and demonstrating effectiveness. All interviewees saw this role as different but complementary to the role of medical staff. There were mixed opinions from service users and professionals on the acceptability of nurse led chemotherapy provision. In defining potential outcomes of nurse-led care, service users described benefits in terms of service and economic outcomes. Professionals saw additional benefits in terms of patient-based outcomes. Professionals and service users expressed the need for appropriate education of nurses for this role and rigorous evaluation of any new service before widespread implementation. CONCLUSIONS: Understanding the perspective of users is imperative when re-designing cancer nursing services. Implementation of nurse-led models in chemotherapy services should be preceded by staff education and followed by systematic evaluation. PMID- 15811104 TI - Newly diagnosed childhood diabetes: a psychosocial transition for parents? AB - AIM: This paper reports a study to gain a new theoretical understanding of parental grief responses and the process of adaptation to a diagnosis of childhood diabetes. BACKGROUND: A diagnosis of childhood (type 1) diabetes is an anxious and distressing event for the whole family. Little is known about the experience of parents of newly diagnosed children as they cope with and adapt to their new situation. Parkes' Theory of Psychosocial Transition proposes that life change events, or 'psychosocial transitions', require people to undertake a major revision of their assumptions about the world. The relevance of this theory to adjusting to a diagnosis of childhood diabetes has not been explored. METHOD: Forty audio taped in-depth interviews were undertaken with 38 parents of 20 newly diagnosed children. The data were subsequently examined using the framework of the Theory of Psychosocial Transition. FINDINGS: Before diagnosis, most parents associated their child's symptoms with normal childhood illnesses. The unexpectedness and speed of the diagnosis left all parents ill-prepared to deal with the situation. Their world suddenly changed, leaving them insecure and uncertain about the future. Diabetes intruded emotionally and practically upon all of their lives. Parents successfully adjusted and adapted their lives and rebuilt a new model of the world to accommodate their child's diabetes. However, this dynamic process has no guaranteed endpoint for parents. CONCLUSIONS: A diagnosis of childhood diabetes leads to a psychosocial transition for parents. The concept of transition provides a logical explanation of parents' responses to loss, and allows increased understanding of the grieving and adaptation processes experienced by parents of children diagnosed with a chronic condition such as diabetes. This knowledge should help health care professionals to assist parents in the period of transition. PMID- 15811105 TI - Body care experienced by people hospitalized with severe respiratory disease. AB - AIM: The paper gives a theoretical account of experiences of assisted personal body care (APBC) in hospitalized patients with severe chronic obstructive pulmonary disease (COPD). BACKGROUND: Body care has been identified as a central but underestimated area of nursing. Hospitalized patients with severe COPD suffer from breathlessness on exertion and are dependent on help with personal body care. Studies have described patient strategies for managing breathlessness and preferences regarding nursing care during hospitalization. Yet the problems that patients can experience because of their inability to manage personal body care by themselves have not previously been explored. This study explored patients' experiences of being assisted with personal body care. METHODS: A grounded theory methodology was used with a convenience sample of 12 patients. Data were gathered from participant observation of sessions of APBC, in-depth interviews after the observed sessions and measurement of breathlessness perceived by patients before and after the sessions. FINDINGS: The patients perceived body care as a significant daily activity that needed to be carried out in order to preserve their integrity. Dependency and breathlessness, however, impeded the performance of body care activities and patients were struggling for self-preservation. They managed APBC by using a threefold strategy of not letting go, coping with dependency, and minimizing the risk of escalating breathlessness. Two dilemmas were identified as being inherent to the strategy. CONCLUSION: Increased knowledge of the complexities involved in providing assistance might improve nurses' ability to facilitate patients in managing APBC. Dependency is a central issue to address in order to support patients' efforts to preserve integrity and resolve dilemmas inherent to the strategy they use. PMID- 15811106 TI - Initiation and continuation of breastfeeding: theory of planned behaviour. AB - AIMS: This paper reports a study investigating new mothers' subjective norms in relation to both breast- and bottle-feeding. The influence of norms on women's infant feeding decisions and the relative influence of social referents at varying degrees of social distance were assessed. BACKGROUND: Increasing breastfeeding initiation and continuation rates is a key challenge for health educators. The perceived influence of other people's views (subjective norms), including the views of women's partners and health care professionals, is an important predictor of infant feeding behaviour. METHODS: Semi-structured questionnaires were administered to 203 new mothers in central and northern Scotland and followed-up by postal questionnaire at 6 weeks. Infant feeding intentions, feeding behaviour at birth and follow-up, behavioural beliefs and subjective norms for both breastfeeding and bottle-feeding were assessed. The data were collected in 1998-1999. RESULTS: Subjective norms were important determinants of initiation and continuation of breastfeeding for breast- and bottlefeeders. Breastfeeders rated close social referents as more in favour of bottle-feeding and more against breastfeeding at follow-up, whereas bottlefeeders' ratings did not change. Partner's and nurses'/midwives' views were an important influence at baseline and follow-up. Breastfeeding 'continuers' perceived their partners as more pro-breastfeeding at 6 weeks. Discontinuers perceived more overall social pressure to bottle-feed. However, sampling limitations may have led to over-representation of the views of breastfeeders at baseline and follow-up. CONCLUSIONS: Nurses and midwives have a crucial role in communicating positive views on breastfeeding to new mothers at different time points. Future interventions to promote breastfeeding could adopt a broad social approach, encouraging positive norms for existing and potential mothers and fathers, families and people in general. PMID- 15811107 TI - Individualized care, quality of life and satisfaction with nursing care. AB - AIMS: This paper reports a study of the maintenance of individualized care from surgical patients' point of view and examines associations between individualized care, patient satisfaction with nursing care, and health-related quality of life. Rationale. There is preliminary evidence that individualized care is effective from patients' point of view, and is associated with improved patient outcomes, such as patient satisfaction and quality of life. However, individualized care has mainly been studied from the vantage-point of nurses' experiences. METHODS: In this cross-sectional, descriptive, correlational study the data were collected with surgical adult patients (n = 279, response rate 93%) in surgical wards in Finland using self-administered questionnaires including the Individualized Care Scales, Patient-Satisfaction Scale, and Finnish versions of the Nottingham Health Profile and EuroQol 5D. Associations between individualized care, satisfaction with care and health-related quality of life were examined. Cronbach's alpha values and item analysis were used to evaluate the psychometric properties of the instruments, especially the Individualized Care Scales. RESULTS: The more often patients felt they received support for individuality through specific nursing interventions, the higher the individuality of care received. Secondly, the more individualized patients regarded their care, the higher the level of reported patient satisfaction with nursing care. However, the correlation between individualized care and health-related quality of life was fairly low, albeit statistically significant. CONCLUSION: Individualized care may produce positive outcomes, such as patient satisfaction. Further research is needed to explore individualized care in relation to health-related quality of life. PMID- 15811108 TI - Unpacking the concept of patient satisfaction: a feminist analysis. AB - AIM: The aim of this paper is to present a feminist critique of the concept of patient satisfaction. BACKGROUND: Fiscal restraint, health care restructuring, shifting demographics, biomedical technological advances, and a significant shortage of health care professionals are stretching health care systems across North America to the breaking point. A simultaneous focus on consumerism and health service accountability is placing additional pressure on the system. The concept of patient satisfaction, with roots in the consumer movement of the 1960s, has both practical and political relevance in the current health care system and is commonly used to guide research related to consumer experiences of health care. Because the quality of health care encounters may lead to treatment seeking delays, patient satisfaction research may be an effective vehicle for addressing this public health issue. However, there is wide agreement that patient satisfaction is an under-theorized concept. Using current conceptualizations of patient satisfaction, we end up all too often producing a checklist approach to 'achieving' patient satisfaction, rather than developing an understanding of the larger issues underlying individual experiences of health care. We focus on the symptoms rather than the problems. DISCUSSION: Without further theoretical refinement, the results of research into patient satisfaction are of limited use. To push forward theoretical development we might apply a variety of theoretical lenses to the analysis of both the concept and the results of patient satisfaction research. Feminism, in particular, offers a perspective that may provoke further refinement of patient satisfaction as a concept. CONCLUSIONS: Without a deeper understanding of the values and beliefs (or the worldview) that informs our approaches to researching patient satisfaction, researchers will be reacting to the most obvious indicators and failing to address the underlying issues related to individual experiences of health care. PMID- 15811109 TI - Giving people a voice: reflections on conducting interviews with participants experiencing communication impairment. AB - AIM: The aim of this paper is to describe the various strategies employed to mitigate communication impairments and to argue that illness experiences may be captured through methods other than audio-tape recordings of spoken words. BACKGROUND: This paper explores issues surrounding the collection of interview data from participants experiencing communication impairments. It derives from a qualitative interview study, which enquired into experiences of living with a percutaneous endoscopic gastrostomy. This is an artificial route created directly into the stomach for long-term delivery of nutrition to people who are unable to take nutrition via the normal route. However, in many cases the physical impairments that led to their requirement for this form of feeding also made speech, and hence interviewing, very difficult. DISCUSSION: Capturing illness experiences from participants with impaired communication requires a reconsideration of both practical strategies and broader methodological issues. In addition to the adaptation of data generation strategies a re-examination of the complexities inherent in the meaning of 'patient experience' is required. This paper explores the utility of verbal and other interactional input from carers to explore further dimensions of this experience. CONCLUSION: Qualitative research that seeks to capture the meanings of illness from the patient's perspective needs to be adapted to embrace different modes of communicating the illness experience. PMID- 15811110 TI - Pressure ulcers: guideline development and economic modelling. AB - AIM: This paper presents the development process for clinical guidelines on the use of pressure-relieving devices (beds, mattresses and overlays), with emphasis on incorporating economic evidence. BACKGROUND: Previous UK guidelines on the use of pressure-relieving devices to prevent pressure ulcers have not considered whether any recommendations made are cost effective. The routine inclusion of cost effectiveness evidence in guidelines is a recent policy development, and there has been little research into its potential role in the guideline process. METHODS: We systematically reviewed the literature to assess both the clinical and cost effectiveness of pressure-relieving devices for prevention of pressure ulcers. Where there was sufficient evidence on the comparative clinical effectiveness between alternative devices, economic modelling was undertaken to assess comparative cost effectiveness. A guideline development group (comprising both clinicians and patient representatives) reviewed all the available evidence to formulate clinical practice guidelines and recommendations for further research. RESULTS: Studies showed that caring for people vulnerable to developing pressure ulcers on high-specification foam mattresses compared with standard hospital mattresses significantly reduced their risk of developing a pressure ulcer. Cost effectiveness modelling indicated that, because of savings accruing through treating fewer pressure ulcers, high-specification foam mattresses are likely to cost less overall and are more effective. The resulting clinical practice guideline was uncompromising on the use of high-specification foam mattresses as the minimum provision in patients vulnerable to pressure ulcers. Significant weaknesses were identified in both the quality and availability of evidence for most of the other pressure relieving devices considered. CONCLUSIONS: Cost effectiveness assessment was an integral part of the guideline development process. It clarified the shortcomings of some of the clinical effectiveness evidence and helped in formulating pragmatic clinical practice recommendations. PMID- 15811111 TI - The impact of international placements on nurses' personal and professional lives: literature review. AB - AIM: This paper presents a critical review of research literature on the impact of international placements on the lives and practice of nurses. BACKGROUND: Health care institutions are progressively more aware of the need to respond to diverse patient populations and cultivate leaders to enrich the nursing profession, both locally and globally. One response has been to establish international exchange programmes for nursing students to give them experience of different cultures and health care systems. METHODS: A search of the literature from 1980 to 2003 using electronic databases was undertaken using the databases CINAHL, ERIC, British Nursing Index, Web of Science, the BIDS Social Science Citation Index and Medline. The keywords used were 'international exchange experience', 'international studies', 'international education', 'international placement(s)', 'exchange programme(s)', combined with 'nurses/nursing', combined with 'evaluation', 'practice', 'education' and/or 'policy'. The papers retrieved used both qualitative and quantitative approaches and were scrutinized for recurring themes. FINDINGS: Nurses reported significant changes in their personal development, perspectives on nursing practice and critical appraisal of health care systems. They also indicated an increased appreciation and sensitivity towards cultural issues and cross-cultural care. Moreover, differences in placement programmes, such as duration, preparation and debriefing were found to have affected the reported overall international placement experience. However, the primary effects of international placements were identified as personal development and transcultural adaptation. CONCLUSION: Students should be exposed to a variety of nursing experiences within the host country. This would give them a broad spectrum for comparisons between cultures, nursing practice and health care delivery in those cultures. Therefore, educational institutions are strongly encouraged to provide opportunities for students to participate in nursing care and education in another country. PMID- 15811112 TI - Stress in carers of individuals with dementia and Community Mental Health Teams: an uncontrolled evaluation study. AB - AIM: The aim of this was to evaluate the effectiveness of interventions provided by a Community Mental Health Team (CMHT) in reducing stress in carers of individuals with dementia. BACKGROUND: The CMHT had been created to working specifically with older people with mental health problems and their carers. Following initial multidisciplinary assessment a range of interventions were provided to both clients and carers according to assessed need. There is an established need for mental health services to focus on the need of carers and the study attempts to see if the interventions provided were useful in reducing carer stress. METHOD: The study used a time series design over a 2-year period on all referrals to the CMHT. All carers of individuals with dementia or clearly identified memory problems were invited to participate and a total of 26 carers consented and participated in all stages of data collection. Data were collected on initial assessment, as well as 3 and 6 months following the initial assessment using the Caregiver Strain Index (CSI). A questionnaire was also administered which collected basic demographic information and details of symptoms demonstrated by the carer's relatives. RESULTS: On initial assessment the mean CSI score for the overall sample was 9.23. The mean CSI reading at 3 months (6.63) and 6 months period (4.12) demonstrated statistically highly significant reductions in carer stress (P = 0.000). A linear stepwise regression analysis of the impact of the different interventions on reductions in the CSI scores showed a statistically significant relationship between respite care and reduction in carer stress (B = 1.705, t = 2.586, P = 0.017). CONCLUSION: The results add support to the role of multidisciplinary community based services for individuals with dementia, offering a range of interventions to both clients and their carers, in reducing carer stress. The authors also argue for the routine use of the CSI in such teams as means of monitoring the well-being of carers as well as evaluating the effectiveness of service delivery. PMID- 15811115 TI - Nucleolar organizer region staining patterns in paraffin-embedded tissue cells from human skin cancers. AB - BACKGROUND: Increased number of nucleoli (nucleolar organizer regions, NORs) with abnormal shapes and sizes, including small dots, has been used as prognostic tools to evaluate tumor proliferation levels and troublesome borderline lesions. In this study, NOR patterns of skin cancers were performed in the search of a valuable prognostic method to complement other histological procedures. METHODS: Paraffin-embedded tumor tissue was obtained from basal and squamous cell carcinomas, cutaneous malignant melanoma, premalignant lesions, and Skmel-28 human melanoma cells. Slices were dewaxed and AgNOR stained. The patterns were scored and submitted for statistical analyses. RESULTS: All types of cancer cells showed variable numbers of abnormally shaped nucleoli and dot-like structures. Only tumor cells presented four or more nucleoli, with or without dots, while 85% of the normal cells had one single NOR without dots. Most data were statistically significant when compared to normal cells. As a whole, squamous cell carcinoma and malignant melanoma tumor cells had less NOR alterations than basal cell carcinoma (BCC) tumor types. CONCLUSIONS: Changes in the number and shape of nucleoli present in malignant cells could be attributed to increased levels on rDNA transcription on cancer cells, besides abnormal remodeling of chromatin, which could disrupt proper nucleoli association. Increased genetic alterations on malignant basal cells could contribute to impair invasive and migration abilities of BCC tumors. PMID- 15811116 TI - Chondrodermatitis nodularis helicis as a marker of internal disease [corrected] associated with microvascular injury. AB - Chondrodermatitis nodularis helicis (CNH) is held to be an idiopathic degenerative process involving the upper dermis of the auricular rim. Chondrodermatitis typically occurs in elderly men where associations with underlying trauma and sun exposure have been postulated as potential inciting triggers. Its association as a marker of systemic disease is not well established. We describe 24 patients with CNH, in whom there were also significant underlying diseases largely associated with vascular injury including those of immune-based etiology and/or conditions which have been previously linked with granuloma annulare, another necrobiotic process of collagen. These patients with concomitant systemic disease were characteristically younger compared to the classic demographics described for CNH. In some cases, chondrodermatitis may represent an ischemic necrobiotic disorder of collagen, potentially defining an important sign of underlying systemic disease in younger patients. PMID- 15811117 TI - Detection of the BRAF V600E mutation in melanocytic lesions using the ligase detection reaction. AB - BACKGROUND: BRAF is a member of the RAF kinase family, and it promotes signaling through the RAS-MAP kinase signal transduction cascade. Research has shown that a majority of melanomas and nevi exhibit an activating V600E (T1799A) mutation in BRAF exon 15. Additional studies of BRAF have demonstrated that the T1799A mutation is absent in uveal melanomas and Spitz nevi. METHODS: The ligase detection reaction (LDR) is a sensitive technique that can identify specific DNA mutations occurring at very low frequency within heterogeneous clinical samples, and it has previously been used to analyze mutations in paraffin-embedded tissue specimens. RESULTS: The LDR can readily detect mutations in DNA extracted from non-microdissected paraffin-embedded sections of common nevi, dysplastic nevi, and melanomas. In addition, this method demonstrated the absence of the V600E (T1799A) mutation within Spitz nevi. CONCLUSION: The LDR is a highly sensitive and specific test for detecting mutations in formalin-fixed tissue. The LDR can be easily adapted to detect any mutation associated with a cutaneous disorder. The absence of the BRAF V600E mutation in Spitz's nevi may serve as a molecular signature to distinguish these lesions from common nevi, dysplastic nevi, and some types of malignant melanoma. PMID- 15811118 TI - Peroxisome proliferator-activated receptors in squamous cell carcinoma and its precursors. AB - BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) mediate several functions that are of interest in carcinogenesis. Although PPARalpha, PPARbeta, and PPARgamma are expressed in multiple human, their expression has not been investigated in non-melanoma skin cancer. METHODS: We performed a retrospective paired immunohistochemical analysis of normal skin, actinic keratosis (AK), and squamous cell carcinoma (SCC) among 35 individuals. Specimens were considered PPAR immunoreactive when 1% or more of the tumor cells showed clear evidence of immunostaining. Cyclooxygenase-2 (COX-2) expression, the fraction of proliferating endothelial cells, and microvessel density were also evaluated in these samples. RESULTS: PPARalpha immunoreactivity was significantly less likely to occur in SCC and AK than in normal skin of each individual. In contrast to PPARalpha, PPARbeta appeared to be upregulated in (pre)malignant skin lesions. For each individual, the likelihood that normal skin, AK, or SCC was immunoreactive against PPARgamma was comparable. COX-2 immunopositivity was significantly associated with PPARbeta and PPARgamma immunoreactivity. No statistical differences were noted for the angiogenesis parameters and PPARalpha, PPARbeta, or PPARgamma expression, except that the microvessel density was significantly higher among PPARbeta-immunoreactive SCCs compared to that among immunonegative SCCs. CONCLUSION: Although further research is warranted, these results suggest that PPAR ligands such as fibrates and thiazolidinediones may have chemoprophylactic properties in skin carcinogenesis. PMID- 15811119 TI - CD10 is expressed in cutaneous clear cell lesions of different histogenesis. AB - BACKGROUND: CD10, the Common Acute Lymphoblastic Leukemia Antigen, is a neutral endopeptidase commonly used as a marker of early B-cell differentiation in the classification of lymphomas. Neoplasms of other histogenesis may express CD10, including renal cell carcinoma. Renal cell carcinoma metastatic to the skin (MRCC) can simulate other more common clear cell lesions in which expression of CD10 has not been described. METHODS: Fifty-two cutaneous clear cell lesions including xanthomas (CX), xanthelasmas (XA), xanthogranulomas (XG), balloon cell nevi (BCN), nodular/clear cell hidradenomas (CCH), and MRCC were examined by immunohistochemistry for the expression of CD10, noting frequency and pattern of labeling. RESULTS: CD10 was expressed in 32/35 of the xanthomatous lesions (CX, XA, and XG), 3/3 MRCC, but only 2/8 BCN and 2/6 CCH. BCN and CCH expressed CD10 in fewer than 10% of the clear cells, whereas all MRCC and most xanthomatous lesions had labeling in greater than 10% (p < 0.001). Xanthomatous lesions exhibited a predominantly membranous pattern of labeling compared to the cytoplasmic pattern of MRCC (p < 0.025). CONCLUSIONS: Cutaneous clear cell lesions of different histogenesis express CD10, limiting its use as a specific diagnostic marker for MRCC. Among other clear cell lesions, however, BCN and CCH have a lower frequency of labeling than does MRCC, and xanthomatous lesions show a membranous pattern compared to the cytoplasmic pattern of MRCC, BCN, and CCH. This latter observation may be indicative of altered protein function or trafficking. PMID- 15811120 TI - Immunoglobulin M predominance in cutaneous lupus erythematosus. AB - BACKGROUND: Direct immunofluorescence (DIF) is a valuable tool in the diagnosis of cutaneous lupus erythematosus (LE). Our goal was to characterize the most frequent immune reactants in the skin biopsies of cutaneous LE and identify the most common immunofluorescence staining patterns. METHODS: DIF results of immunoglobulin G (IgG), IgA, IgM, C3, and fibrinogen from 199 patients between 1989 and 1998 were retrospectively analyzed. Confirmatory clinical and serological diagnosis of LE subtype was available for 95 patients. Intensity of staining was ranked from 0 to 4+ but only included as significant if >/=2+. Laboratory values were gathered and analyzed for all patients who had distinct granular immune deposition for Ig and/or complement. RESULTS: The most commonly detected individual Ig was IgM in 149 (75%) specimens. IgM and C3 combination was the most common pair expressed with 98 (49%) specimens. The most common triplet was IgM, C3, and fibrinogen in 63 (32%) specimens. The most common quadruplet was the combination of IgG, IgM, C3, and fibrinogen in 42 (21%) specimens. All the five immunoreactants were detected in only 25 (13%) specimens. Systemic LE patients had a higher percent of abnormal laboratory values compared to discoid LE (DLE) and subacute LE (SCLE) patients (p = 0.02). Fibrinogen staining was found to be significantly higher in DLE patients and lowest in SCLE patients (p = 0.05). CONCLUSIONS: This study demonstrates a marked predominance of IgM +/- C3 in cutaneous LE. When used in conjunction with other data, DIF is an extremely powerful tool in the routine evaluation of the LE patient. Our report emphasizes the importance of IgM expression in the diagnosis of LE by DIF and how positive staining with multiple conjugates can raise its sensitivity. PMID- 15811121 TI - Comparison of benign keratoses using p53, bcl-1, and bcl-2. AB - While cell-cycle markers have been used to differentiate benign vs. malignant lesions and to classify malignant lesions, benign keratoses have not been well studied using such markers. We hypothesized that inflammation or irritation of benign keratoses may be related to a shift in the cell cycle. We compared the immunohistochemical staining patterns of 10 seborrheic keratoses (SKs), 10 inflamed seborrheic keratoses (iSKs), and 10 inverted follicular keratoses (IFKs) using antibodies to p53, bcl-1, and bcl-2. Staining with antibodies to p53 was slightly increased in IFKs compared with iSKs or non-inflamed seborrheic keratoses. Bcl-1 staining was similar in all lesions. A population of bcl-2 positive dendritic cells was seen within the epidermal portion of IFKs. Keratinocyte bcl-2 staining was significantly higher in SKs compared with the other two keratoses. Bcl-2 may be increased in SKs as an anti-apoptotic mechanism. PMID- 15811122 TI - Fatal subcutaneous panniculitis-like T-cell lymphoma with interface change and dermal mucin, a dead ringer for lupus erythematosus. AB - We report a 48-year-old man who presented with ulcerated plaques and nodules of the lower extremities. Skin biopsies revealed a dense lymphocytic infiltrate involving the dermis and the subcutis in a lobular and septal pattern. No overt cytological atypia was present. Notably, several features resembling lupus erythematosus were present, including vacuolar interface change and abundant dermal mucin deposition. The patient developed pulmonary nodules, and a lung biopsy showed a perivascular and interstitial lymphoid infiltrate without overt atypia. The cutaneous and pulmonary lymphoid infiltrates showed similar immunohistochemical profiles: CD3(+) CD4(-) CD8(+/-) CD56(+). Monoclonal rearrangements of the T-cell receptor gamma gene with similar migration patterns were identified from both locations. The patient developed fatal hemophagocytic syndrome, involving liver, spleen, lymph nodes, and bone marrow. This case is one amongst the rare reports of subcutaneous panniculitis-like T-cell lymphoma with systemic involvement. PMID- 15811123 TI - Subcutaneous panniculitic T-cell lymphoma in a cardiac allograft recipient. AB - BACKGROUND: Post-transplant lymphoproliferative disorder (PTLD) is the third leading cause of death in heart transplant patients beyond the immediate peri operative period (Ouseph R, Denny DM, Erbeck KM. J Am Soc Echocardiogr 1998; 11: 758; Armitage JM, Kormos RL, Stuart RS, et al. J Heart Lung Transplant 1991; 10: 877; Swinnen LJ, Mullen M, Carr TJ, et al. Blood 1995; 86: 3333; Ying AJ, Myerowitz D, Marsh WL. Ann Thorac Surg 1997; 64: 1822). The majority of PTLD cases are of B-cell origin whereas T-cell neoplasms have been reported as rare, aggressive, and late complications of solid-organ transplantation (Fatio R, Sutsch G, Mayer K, et al. Transplant Proc 1998; 30: 1118). CASE REPORT: A 50-year old cardiac allograft heart transplant patient presented with subcutaneous nodules involving his trunk and extremities. RESULTS: Light microscopy revealed features characteristic of subcutaneous panniculitic-like T-cell lymphoma. Immunohistochemical analysis showed expression for CD45RO, TIA-1, and focal CD3 positivity by tumor cells. Flow cytometry performed on a subsequent subcutaneous nodule demonstrated an abnormal T-cell population with expression of CD3, CD8, CD56, and T-cell receptor alpha-beta, and no expression of CD4. T-cell gene rearrangement studies revealed a clonal population of cells with a bi-allelic gene rearrangement. CONCLUSION: We report a case of an unusual subtype of PTLD in a cardiac allograft recipient. PMID- 15811124 TI - Erythroderma with lichenoid granulomatous features induced by erythropoietin. AB - The increasing use of new drugs in cancer therapy, especially growth factors, hormones, and chemotherapies resulted in several reports of unusual skin eruptions. We studied a patient with erythroderma who had received erythropoietin because of myeloma with tumor anemia. The histological features were characterized by a lichenoid, focally granulomatous infiltrate with predominance of histiocytes. It is important for dermatopathologists to recognize this interesting pattern induced by erythropoietin. PMID- 15811125 TI - Myxoinflammatory fibroblastic sarcoma of the neck. AB - BACKGROUND: Myxoinflammatory fibroblastic sarcoma (MIFS), also named inflammatory myxohyaline tumor of distal extremities with virocyte or Reed-Sternberg cells, is a rare tumor typically presenting as a painless mass in the extremities. PATIENTS: We present an unusual case of MIFS presenting as a subcutaneous neck mass. This is the first reported case of MIFS presenting in the neck. RESULTS: Therefore, this lesion must be considered in the differential diagnosis for painless subcutaneous masses presenting not only in the distal extremities, but also in the neck. CONCLUSION: MIFS has only recently been recognized. The differential diagnosis for MIFS is broad, and it can often be mistaken for several different inflammatory and neoplastic processes, which may require different treatment. PMID- 15811126 TI - Two cases of angiomyxolipoma (vascular myxolipoma) of subcutaneous tissue. AB - Angiomyxolipoma (vascular myxolipoma) is a recently described rare variant of lipoma, four cases of which have been reported to date. Microscopically, the lesion consists of adipose tissue without lipoblasts, extensive myxoid areas, and numerous blood vessels. The main differential diagnosis of this lesion is myxoid liposarcoma, and other adipocytic lesions such as myxolipoma, myxoid spindle cell lipoma should be included. We report two cases of angiomyxolipoma located in the subcutaneous tissue of the forearm and the wrist. PMID- 15811127 TI - How far can we trust cell cycle biomarkers for the identification of high-risk melanoma patients? PMID- 15811128 TI - Angiomatoid cellular blue nevus: a variant of blue nevus with an angioma-like appearance. PMID- 15811129 TI - Selective growth and expansion of human corneal epithelial basal stem cells in a three-dimensional-organ culture. AB - We report on a three dimensional (3D)-organotypic culture in vitro for selective growth and expansion of human corneal epithelial stem cells. Limbal corneal explants were cultured on porous collagen sponges submerged in Epilife medium containing 10% fetal bovine serum. The fragments were analyzed by immunohistochemistry for the expression and distribution of a spectrum of corneal epithelium markers: p63, CK-19, CK-3, Ki-67, pan-cytokeratins and vimentin. Early in culture the epithelium began to exfoliate losing its differentiated high-zone layers into the medium, maintaining only basal and few parabasal cells (mostly both p63 and CK-19 positive), which had remained attached to the specimen. After 14 days a new epithelium was formed displaying an increasing prominence of basal and suprabasal cells that, sliding onto the whole explant, showed the tendency to underlay stromal tissue and infiltrate into the underlaying sponge. After 21 days, sponge and fragments were incubated with trypsin-EDTA and dispersed epithelial cells were pipetted on a feeder monolayer of mitomycin-c-treated murine NIH.3T3 fibroblasts. Colonies of undifferentiated epithelial cells (p63, CK-19 and Ki-67 positive, CK-3 negative) were obtained: their cells, if seeded onto a collagen matrix containing embedded primary human corneal fibroblasts as feeder, provided the basic building blocks for reconstructing in vitro a 3D multilayered corneal epithelium. PMID- 15811130 TI - Isolation of two human fibroblastic cell populations with multiple but distinct potential of mesenchymal differentiation by ceiling culture of mature fat cells from subcutaneous adipose tissue. AB - Adipose tissue is a source of adult multipotent stem cells that can differentiate along mesenchymal lineage. When mature fat cells obtained from human subcutaneous adipose tissue were maintained with attachment to the ceiling surface of culture flasks filled with medium, two fibroblastic cell populations appeared at the ceiling and the bottom surface. Both populations were positive to CD13, CD90, and CD105, moderately positive to CD9, CD166, and CD54, negative to CD31. CD34, CD66b, CD106, and CD117, exhibited potential of unlimited proliferation, and differentiated along mesenchymal lineage to produce adipocytes, osteoblasts, and chondrocytes. The population that appeared at the ceiling surface showed higher potential of adipogenic differentiation. These observations showed that the cells tightly attached to mature fat cells can generate two fibroblastic cell populations with multiple but distinct potential of differentiation. Since enough number of both populations for clinical transplantation can be easily obtained by maintaining fat cells from a small amount of subcutaneous adipose tissue, this method has an advantage in preparing autologous cells for patients needing repair of damaged tissues by reconstructive therapy. PMID- 15811131 TI - Epithelial-mesenchymal interactions allow for epidermal cells to display an in vivo-like phenotype in vitro. AB - We here report that preservation of the basic epithelial-mesenchymal interactions allows for highly complex ex vivo function of epidermal cells. The approach taken is based on the preparation of organ fragments that preserve the basic epithelial/mesenchymal interactions but also ensure appropriate diffusion of nutrients and gases to all cells. Human and mice keratinocytes in such organ fragments, remain viable, proliferate and express epidermal-specific gene products when cultured in serum-free medium without added growth factors, for several weeks in vitro. When implanted into syngeneic animals they remain viable, become vascularized and continue to function and transcribe tissue-specific gene products for several months. Such fragments allow primary cells ex vivo to preserve most of the functional attributes of the in vivo system. Clearly, the effect of the extracellular matrix is critical in this system in order for the cells to proliferate and differentiate ex vivo. We are not aware of any other system which allows for localized expression of epidermal-specific genes ex vivo for significant periods in culture in defined serum-free medium. PMID- 15811132 TI - Tumor-host interactions contribute to the elevated expression level of alpha1 antichymotrypsin in metastatic breast tumor xenografts. AB - We investigated alpha1-antichymotrypsin (ACT) gene expression in xenograft tumors generated by two isogenic human breast cancer cell lines derived from the same parent, MDA-MB-435, which display opposite metastatic behaviors. Microarray and real-time PCR experiments showed an overexpression of this serine protease inhibitor in the metastatic tumors (M-4A4T) and its derived metastases (M4-Mets) compared with the weakly metastatic tumors (NM-2C5T), and its release into the blood was confirmed by western-blotting. However, functional assays in vivo using genetically engineered tumor cells demonstrated that ACT up-regulation was not, by itself, responsible for the metastatic phenotype. We also made observations that ACT gene regulation was sensitive to tumor-host interactions: inoculation of these lines into the mouse mammary gland greatly increased ACT production and accentuated the intrinsic difference observed when they are cultured in vitro. Sensitivity of tumor cells to their environment was further analyzed by in vitro experiments, which demonstrated that a purified ECM environment and soluble components from normal host mammary cells were both able to significantly promote ACT expression. In addition, we took advantage of the xenogeneic nature of the model to measure ACT expression by the host cells (mouse) and the tumor cells (human) within the neoplasm using species-specific primers in real-time PCR experiments. It was found that the presence of tumor cells, irrespective of their metastatic capabilities, induced local ACT production by host cells at the primary and secondary tumor sites. Thus, this work indicates that there is a specific association of ACT overexpression with the metastatic phenotype in our breast cancer metastasis model. Moreover, because of the xenogeneic nature of our system, we were able to provide evidence of tumor-host reciprocal regulation of ACT production. PMID- 15811133 TI - Assembly and breakdown of Cajal bodies in accessory nuclei of Hymenoptera. AB - In some species of insects, oocytes have vesicular organelles, termed accessory nuclei (ANs). The ANs form by budding off from the nuclear envelope of the oocyte and are filled with translucent matrix containing dense inclusions. One type of these inclusions contains coilin and small nuclear ribonucleoproteins (snRNPs) and is homologous to Cajal bodies. We describe the early events in the morphogenesis of Cajal bodies in the ANs (ANCBs) of the common wasp, Vespula germanica, and show that they contain survival of motor neurons (SMN) protein. We present evidence that in the wasp, ANCBs form by the gradual accumulation of aggregates composed of SMN and small nuclear RNAs. We also show that ANCBs break down and disperse within the ANs as the ANs, which initially surround the oocyte nucleus, localize to the oocyte cortex. The components of dispersed ANCBs are retained within ANs until the end of oogenesis, which suggests that their function may be required at the onset of embryonic development. Because the morphology and behavior of ANs and their Cajal body-like inclusions are conserved in two other hymenopteran species, these features might be characteristic of all hymenopterans. PMID- 15811135 TI - Obesity in 2005 and in DOM. PMID- 15811134 TI - Gli1 is not required for Pdgfralpha expression during mouse embryonic development. AB - Pdgfra is expressed in the mesenchyme of multiple organs during embryonic development and Pdgfralpha is involved in cell proliferation, differentiation, migration, and apoptosis in many tissues. A fine-tuned regulation of gene transcription is required to achieve these effects. To investigate if the Shh signaling pathway is involved in the tightly regulated Pdgfra expression during embryogenesis, we systematically compared Gli1 and Pdgfralpha mRNA expression patterns in vivo from mouse embryonic day 9.5 to 14.5. We found that an initial partly overlapping expression of Gli1 and Pdgfralpha in the mesenchyme of foregut and somites was changed to different expression patterns when the mesenchyme differentiated into specialized structures such as intestinal villi and chondrocytes. Gli1 and Pdgfra were also expressed differently in the developing lung, heart, central nervous system, skin, tooth, and eye. Importantly, neither Pdgfralpha mRNA patterns nor levels were altered in Ihh mutant embryos although Gli1 and Ptc mRNA levels were dramatically reduced. Our results demonstrate that Gli1 is not required to induce Pdgfra expression during embryonic bone development, and are consistent with previous findings that Pdgfralpha and Hh pathways serve different functions in, e.g., bone, gut, and lung development. However, we cannot exclude the possibility that Glis can have more complex regulatory effects on Pdgfra gene activity, nor can we exclude such effects in pathological conditions. PMID- 15811136 TI - Letrozole normalizes serum testosterone in severely obese men with hypogonadotropic hypogonadism. AB - BACKGROUND: Morbid obesity is associated with increased estradiol production as a result of aromatase-dependent conversion of testosterone to estradiol. The elevated serum estradiol levels may inhibit pituitary LH secretion to such extent that hypogonadotropic hypogonadism can result. Normalization of the disturbed estradiol-testosterone balance may be beneficial to reverse the adverse effects of hypogonadism. AIM: To examine whether aromatase inhibition with Letrozole can normalize serum testosterone levels in severely obese men with hypogonadotropic hypogonadism. PATIENTS AND METHODS: Ten severely obese men, mean age 48.2 +/- 2.3 (s.e.) years and body mass index 42.1 +/- 2.6 kg/m(2), were treated with Letrozole for 6 weeks in doses ranging from 7.5 to 17.5 mg per week. RESULTS: Six weeks of treatment decreased serum estradiol from 120 +/- 20 to 70 +/- 9 pmol/l (p = 0.006). None of the subjects developed an estradiol level of less than 40 pmol/l. LH increased from 4.5 +/- 0.8 to 14.8 +/- 2.3 U/l (p < 0.001). Total testosterone rose from 7.5 +/- 1.0 to 23.8 +/- 3.0 nmol/l (p < 0.001) without a concomitant change in sex hormone-binding globulin level. Those treated with Letrozole 17.5 mg per week had an excessive LH response. CONCLUSION: Short-term Letrozole treatment normalized serum testosterone levels in all obese men. The clinical significance of this intervention remains to be established in controlled, long-term studies. PMID- 15811137 TI - Metabolic risk factors in formerly obese women--effects of a pronounced weight loss by gastric band operation compared with weight loss by diet alone. AB - AIM: The aim of the present study was to evaluate differences in the metabolic risk profile in formerly obese women, in whom a significant weight loss was obtained by Swedish adjustable gastric band (SAGB) operation or by diet alone. METHODS: A total of 40 patients (24 after SAGB and 16 after diet) participated in the study. Clinical data, including body mass index (BMI), waist-to-hip ratio, body fat content and blood pressure values, as well as laboratory results [fasting glucose, insulin, homeostasis model assessment (HOMA) index, leptin, lipid values and markers of subclinical inflammation] were evaluated before a planned abdominoplastic operation. RESULTS: Patients in the SABG group had lost a significantly greater amount of weight (52.7 +/- 10.0 kg) compared with the diet group (20.0 +/- 11.5 kg; p < 0.001), and the percent excess weight loss was 69.1 +/- 11.4 in the SAGB group and 54.5 +/- 17.7 (p < 0.040) in the diet group. Before the abdominoplastic operation neither the mean BMI nor the percentage of fat mass revealed a significant difference between the groups. Fasting insulin (6.1 +/- 3.0 microU/ml) and the HOMA index (1.4 +/- 0.7) as a measure of insulin resistance were significantly lower in the SAGB than in the diet group (fasting insulin: 8.2 +/- 3.8 microU/ml; p < 0.048; HOMA index: 2.0 +/- 1.0; p < 0.031). Swedish adjustable gastric band patients showed significantly lower plasma leptin levels (9.4 +/- 10.8 ng/ml) than the dietary-treated patients (13.9 +/- 9.6 ng/ml; p < 0.014), while tumour necrosis factor-alpha serum levels were increased in the SAGB group (17.6 +/- 7.3 pg/ml) compared with the diet group (11.9 +/- 0.49 pg/l; p < 0.048). CONCLUSIONS: The extensive weight loss in formerly obese women after SAGB operation was paralleled by a favourable metabolic profile indicating a higher degree of insulin sensitivity than in women after a successful, but less pronounced weight loss by diet alone. PMID- 15811138 TI - The effect of diets high in protein or carbohydrate on inflammatory markers in overweight subjects. AB - AIM: Increased levels of inflammatory markers have been associated with weight gain and cardiovascular disease. The aim of the present study was to investigate the effect of diets high in either carbohydrate or protein on the inflammatory markers C-reactive protein (CRP), haptoglobin and transferrin in plasma after weight loss. METHODS: Fifty overweight subjects [age: 18-56 years, body mass index (BMI): 27-34] were randomly assigned to an ad libitum, fat-reduced diet (30% of energy, E%), either high in protein and low in carbohydrate (25 E and 45 E% respectively) or high in carbohydrate and low in protein (58 E and 12 E% respectively), during 6 months of strictly controlled dietary intervention with dietary counselling. RESULTS: An average reduction of 25% in CRP and an average increase of 20% in haptoglobin and transferrin were seen in both groups, however, these changes were not significant. In cross-sectional analyses after the intervention CRP was associated with fat mass (r = 0.323, p = 0.03), and the changes in CRP were associated with various indices of body fatness (Deltabody weight r = 0.346, p = 0.02). Changes in body fatness were positively associated with Deltatransferrin (r = 0.344, p = 0.02) and nearly significantly associated with Deltahaptoglobin (r = 0.271, p = 0.07) after 6 months. Multiple regression analysis showed no associations between dietary protein and carbohydrate content and serum CRP, haptoglobin or transferrin concentrations, and this remained unaltered after adjustment for weight change. CONCLUSION: Dietary carbohydrate/protein ratio has no effect on inflammatory markers, but the study confirmed that body fatness is positively associated with levels of serum CRP. PMID- 15811139 TI - Abdominal obesity predicts declining insulin sensitivity in non-obese normoglycaemics: the Insulin Resistance Atherosclerosis Study (IRAS). AB - AIM: Cross-sectional studies have demonstrated a relationship between obesity and insulin sensitivity (S(I)); however, there is a lack of evidence from longitudinal studies. METHODS: The Insulin Resistance Atherosclerosis Study (IRAS) estimated S(I) (x10(-4)/min.microU/ml) directly using a frequently sampled intravenous glucose tolerance test with minimal model analysis in 504 normoglycaemic subjects. Partial correlation coefficients (r) were calculated to compare the relationship of change in S(I) from baseline to 5 years later (DeltaS(I)) with baseline waist circumference (waist) as a measure of abdominal obesity and body mass index (BMI) as a measure of overall obesity. Mean DeltaS(I) was -1.06 (SD = 1.85). RESULTS: Higher baseline waist (r = -0.16; p = 0.0005), but not BMI (r = -0.005; p = 0.91), was associated with (-) DeltaS(I) in models including sex, ethnicity, clinical centre and baseline S(I), BMI, waist, age and physical activity. The waist-DeltaS(I) relationship differed across the levels of baseline BMI, being significant only in normal weight (r = -0.21) and overweight subjects (r = -0.16), but not in obese subjects. DeltaS(I) was correlated with a 5-year change in either obesity measure (Deltawaist: r = -0.22 and DeltaBMI: r = 0.20; p = 0.0001). CONCLUSIONS: Among non-diabetics, waist circumference was a strong predictor of declining S(I) among lean subjects, a modest predictor among overweight subjects, but was not predictive among obese individuals. Waist circumference should be considered, in addition to BMI, when identifying individuals at high risk of diabetes or the insulin resistance syndrome. PMID- 15811140 TI - Diazoxide-mediated insulin suppression in obese men: a dose-response study. AB - BACKGROUND: It has been suggested that diazoxide (DZX)-mediated insulin suppression may be useful to promote weight loss in obese subjects. AIM: To assess the DZX-dose range that is safe to use in obese hyperinsulinaemic men. METHODS: Assessment of DZX efficacy and safety was based on plasma glucose and insulin responses to a standardized 500-kcal breakfast, taken on the sixth day of treatment. Basic information regarding the potential efficacy of DZX treatment was first evaluated in an open-label study in five non-obese men. Subsequently, a double-blind, randomized, placebo-controlled study was performed in 12 obese but otherwise healthy men, comparing placebo treatment with DZX in doses of 50, 75 and 100 mg three times daily for 6 days. RESULTS: In non-obese subjects, DZX 50 mg decreased peak insulin levels by +/-28% and raised peak glucose concentration from 7.1 +/- 0.6 to 7.8 +/- 0.6 mmol/l (p < 0.05). DZX 100 mg reduced peak insulin levels by 45% and caused a rise in peak glucose levels from 7.1 +/- 0.6 to 9.0 +/- 0.9 mmol/l (p < 0.05). In obese men, the 50 and 75 mg doses had no significant effects on glucose or insulin levels. DZX 100 mg reduced the peak insulin levels and insulin area under the curve by +/-20% (p < 0.05) but did not affect fasting or postprandial glucose levels. The relatively limited insulin suppressive effects in obese subjects were attributed to the low plasma DZX levels that were achieved in this group. For comparable doses, plasma DZX levels were about 30% lower in obese than in non-obese men. Plasma DZX levels were highly dependent on dose (p < 0.001) and body weight (p < 0.001). Ninety-two percent of the total variability in DZX levels was explained by these two parameters. CONCLUSION: DZX-mediated insulin suppression is dose dependent in normal and in obese men. However, the efficacy of DZX is much less in obese than in non-obese subjects. This is attributed to weight-dependent differences in distribution volume that lead to markedly lower plasma DZX levels in obese subjects. Weight-adjusted doses will be needed to achieve biologically effective plasma DZX levels. Extrapolation of the data suggests that effective insulin suppression in obese men will at least require a daily dose of 3.2-4.2 mg/kg. PMID- 15811141 TI - Effects of sibutramine on abdominal fat mass, insulin resistance and blood pressure in obese hypertensive patients. AB - OBJECTIVE: The objective of this study is to assess the effects of sibutramine on body weight, body fat distribution, insulin resistance, plasma leptin, lipid profile and blood pressure profiles in hypertensive obese patients. METHODS: Eighty-six central obese hypertensive patients (BMI = 39 +/- 5 kg/m(2), 84% of women, 48 +/- 8.5 years old) were placed on a hypocaloric diet and placebo therapy for 4 weeks. They were then randomized to receive sibutramine (10 mg) or placebo for 24 weeks. Both, before therapy and at the end of the study, the waist and hip circumferences were measured and the waist/hip ratio (WHR) was calculated; abdominal ultrasonography was performed in order to estimate the amount of subcutaneous fat (SF) and visceral fat (VF), and the visceral/subcutaneous ratio. Beyond HOMA-r, another insulin resistance index (IRIp) was calculated by means of the formula: peak of blood glucose after oral glucose load x plasma insulin level/10(4). Fasting plasma leptin and lipid levels were also determined. RESULTS: Sibutramine induced greater weight reduction than placebo (6.7 vs. 2.5%, p < 0.001). Reductions in WHR (0.97 +/- 0.08 vs. 0.94 +/- 0.07, p < 0.01), IRIp (0.11 +/- 0.07 vs. 0.09 +/- 0.06 mmol mu/l(2)) and VF (6.4 +/- 2.4-6.0 +/- 2.4 cm, p < 0.01) were observed only with sibutramine. Plasma leptin decreased with placebo (24 +/- 15 vs. 18 +/- 10 UI/l, p < 0.01), but not with sibutramine (18.8 +/- 8.4 vs. 18.2 +/- 13.2 UI/l). No clinically significant change in lipid profile was observed in both groups. Moreover, office and 24-h blood pressure values did not change during placebo or sibutramine therapy, whereas a significant increase in office heart rate, from 78.3 +/- 7.3-82 +/- 7.9 b.p.m., p = 0.02, was observed with sibutramine. CONCLUSIONS: Sibutramine therapy induced greater body weight loss than placebo in hypertensive obese patients. This was associated with WHR reduction, decreases in VF and insulin resistance. The maintenance of leptin levels during sibutramine therapy may be important to avoid weight recovery, although this finding must be confirmed by other prospective studies. PMID- 15811142 TI - Effect of orlistat on cardiovascular disease risk in obese adults. AB - AIM: The aim of this study is to compare the effect of orlistat vs. placebo on the predicted 10-year cardiovascular disease (CVD) risk in obese people with one or more cardiovascular risk factors treated for 12 months, in conjunction with a fat-reduced, but otherwise ad libitum, diet. METHODS: A double-blind, randomized, placebo-controlled, parallel study was performed in conjunction with a fat reduced diet and physical activity advice for 1 year. Participants (n = 339) from eight centres in Australia and New Zealand were randomized to either orlistat (120 mg) three times daily (n = 104 women, 66 men; mean +/- s.d. age = 52.0 +/- 7.5 years, body mass index (BMI) = 37.6 +/- 5.1 kg/m(2)) or placebo three times daily (n = 89 women, 80 men; age = 52.5 +/- 7.4 years, BMI = 38.0 +/- 4.9 kg/m(2)). The primary efficacy criterion was the 10-year risk of developing CVD calculated from the Framingham equation. Secondary efficacy criteria were body weight, waist circumference, blood pressure and serum concentrations of triglycerides, cholesterol (total, LDL and HDL), glucose, insulin and glycated haemoglobin and quality of life. RESULTS: There was no difference in the change in 10-year CVD risk between orlistat and placebo groups over 1 year. The orlistat group, however, had significant favourable changes in many of the individual CVD risk factors (total cholesterol, LDL-cholesterol, glucose, glycated haemoglobin, insulin, body weight and waist circumference) and one of the domains of quality of life measured by means of the SF-36 questionnaire (vitality), compared to the placebo group. Significant reductions in medication use for hypertension and diabetes were observed in the orlistat group, compared to those in placebo, but there were no significant differences in medication use for blood lipids. CONCLUSIONS: Orlistat may have reduced CVD risk, as judged by the favourable changes in individual risk factors and reductions in medication use, but the method used in order to measure absolute CVD risk in this study (Framingham CVD equation) was not sensitive enough to detect the changes in this relatively low risk group (approximately 10% of risk of a CVD event over 10 years). PMID- 15811143 TI - Clinical implications of N epsilon-(carboxymethyl)lysine, advanced glycation end product, in children and adolescents with type 1 diabetes. AB - AIM: The aim of this study was to investigate the relationship between serum levels of the glycoxylation product N(epsilon)-(carboxymethyl)lysine (CML) and development of chronic diabetic complications and degree of diabetic control in children and adolescents with type 1 diabetes. METHODS: The serum levels of CML were measured in 87 patients with uncomplicated type 1 diabetes mellitus (12.7 +/ 4.6 years of age) and in seven patients with background retinopathy, microalbuminuria or neuropathy (18.2 +/- 5.2 years of age) and compared with those in 64 normal control subjects (12.6 +/- 5.2 years of age). The mean durations of diabetes in uncomplicated and complicated patients were 5.0 +/- 3.4 years (0.1-14 years), and 8.6 +/- 5.0 years (3.1-18 years), respectively. The serum levels of CML were measured by enzyme-linked immunosorbent assay using a monoclonal anti-CML antibody (6D12). RESULTS: The serum levels of CML were significantly higher in the patient group than those in the control group; 0.85 +/- 0.37 (0.37-1.93) U/ml vs. 0.56 +/- 0.23 (0.15-1.05) U/ml (p < 0.001) and significantly higher in the patient group with chronic complications than those in patient group without chronic complications; 1.06 +/- 0.39 (0.72-1.78) U/ml vs. 0.83 +/- 0.36 (0.37-1.93) U/ml (p < 0.05). Weak, but statistically significant relationship between CML levels and haemoglobin A(1c) levels at the measurement of CML was observed (r = 0.29, p < 0.05). CONCLUSIONS: Our data are suggesting that higher serum levels of CML are involved in the development of chronic diabetic complications, and serum levels of CML reflect the degree of diabetic control for a long duration in type 1 diabetic children and adolescents. PMID- 15811144 TI - Comparison of air displacement plethysmograph and bioelectrical impedance for assessing body composition changes during weight loss in Japanese women. AB - OBJECTIVE: To investigate the difference between air displacement plethysmograph (ADP) and bioelectrical impedance (BI) for assessing body composition changes during weight loss in Japanese women. DESIGN: Cross-sectional and longitudinal investigation study. SUBJECTS: In a first cross-sectional study, we used data of 7380 Japanese women aged 14-83 years [body mass index (BMI): 22.6 +/- 3.7 kg/m(2)] at baseline. Among the participants, we used data of 54 women (BMI: 25.9 +/- 3.9 kg/m(2)) for a 5-month follow-up study. MEASUREMENTS: Body fat percentage was evaluated by ADP and BI, and body fat (kg) and lean body mass (kg) were calculated. Anthropometric parameters, such as height, weight, BMI, waist circumference, hip circumference and waist hip ratio, were also measured. In a second longitudinal study, they were also investigated before and after the study. RESULTS: In a first cross-sectional study, body fat percentage was 31.2 +/ 7.0% in ADP and 25.7 +/- 6.8% in BI. A significant relationship was observed between ADP and BI (r = 0.761, p < 0.01). In a second longitudinal study, body weight was significantly reduced. There were significant method (ADP vs. BI) and time (pre and post) effects but no interactions in assessing body fat percentage. There was a significant relationship between ADP and BI in body fat percentage, body fat and lean body mass. Delta body fat percentage (r = 0.386, p < 0.01) and delta body fat (r = 0.556, p < 0.01) measured by ADP were weakly correlated with those measured by BI. However, delta lean body mass measured by ADP was not correlated with that measured by BI. CONCLUSION: There was a discrepancy between ADP and BI for assessing body composition changes in Japanese women. ADP is preferentially recommended for evaluating body composition. PMID- 15811145 TI - Effects of cerivastatin on forearm vascular responses, blood pressure responsiveness and ambulatory blood pressure in type 2 diabetic men. AB - OBJECTIVE: The objective of the study was to investigate the effects of cerivastatin therapy on forearm endothelial dependent acetylcholine (ACH) and independent (nitroprusside) vasodilator responses, blood pressure (BP) responses to intravenous infusions of angiotensin II (AII) and noradrenaline (NA) and on 24 h ambulatory BP recordings in type 2 diabetic men. DESIGN: Eleven type 2 diabetic men aged 59 +/- 9 years with total cholesterol levels of 5.0 +/- 1.26 mmol/l, triglycerides of 2.23 mmol/l and high-density lipoprotein cholesterol levels of 1.24 mmol/l completed a double-blind, randomized, crossover trial comparing 8 weeks of cerivastatin therapy (800 microg of nocte) with placebo. Forearm vascular resistance (FVR) responses to intrabrachial-arterial infusions of ACH (3 24 microg/min), nitroprusside (2-16 microg/min), the nitric oxide(NO) synthase inhibitor l-nitro-mono-methyl arginine (l-nmma) (8 micromol/min), ACH during l NMMA infusion and BP responses to intravenous infusions of AII (12.5-50 ng/min) and NA (20-400 ng/min) were measured at the end of each treatment period. Twenty four-hour ambulatory BP recordings were also performed. RESULTS: FVR responses to ACH during l-NMMA infusion were significantly (p = 0.026) greater during cerivastatin than during placebo therapy. In contrast, FVR responses to ACH in the absence of NO synthase inhibition did not differ significantly between cerivastatin and placebo therapies (p = 0.81). FVR increased by 31.4 +/- 57.3% in response to l-NMMA infusion during cerivastatin therapy compared with 6.1 +/- 41.2% during placebo therapy (p = 0.20). FVR responses to nitroprusside did not differ between cerivastatin and placebo therapies (p = 0.28), nor did BP responses to AII (systolic BP, p = 0.99; diastolic BP, p = 0.98) or NA (systolic BP, p = 0.21; diastolic BP, p = 0.48). Mean 24-h BP was similar during cerivastatin (123 +/- 10 or 70 +/- 7 mmHg) and placebo therapies (129 +/- 11 or 74 +/- 7 mmHg) (systolic BP, p = 0.26; diastolic BP, p = 0.41). CONCLUSION: Cerivastatin increases FVR responses to ACH in type 2 diabetic men with mild dyslipidaemia but only following NO synthase inhibition. This may indicate an improvement in endothelium-derived hyperpolarizing factor-mediated responses. PMID- 15811146 TI - Predictors of glycaemic control in indigent patients presenting with diabetic ketoacidosis. AB - AIM: To derive predictors of good glycaemic control in patients presenting with diabetic ketoacidosis (DKA) followed prospectively in a specialized clinic. METHODS: One hundred and sixty-one adult patients were admitted during a 31-month period and followed for at least 12 months. After 1 year, the patients were classified into three groups: good control (GC) (HbA1c < or = 7%), intermediate control (IC) (HbA1c 7-9%) and poor control (PC) (HbA1c > 9%). Characteristics of patients in the three groups were compared both at baseline and during follow-up. RESULTS: At 12 months, 36% of the patients were classified as GC, 27% as IC and 37% as PC. GC patients had higher fasting serum C-peptide levels 0.7 +/- 0.54 compared to 0.38 +/- 0.29 and 0.16 +/- 0.21 nmol/l, respectively, for the IC and PC patients (p < 0.0001). A higher proportion GC patient had a C-peptide level greater than 0.33 nmol/l than that for IC and PC patients (86, 61 and 19%, respectively; p < 0.0001). Exogenous insulin was safely discontinued in 50, 30 and 3% of patients, respectively, in the GC, IC and PC groups (p < 0.0001). Compliance with life-style interventions was higher in the GC than that in IC and PC patients (87, 41 and 5%, respectively; p < 0.0001). In the logistic regression analysis, predictors of good glycaemic control were having baseline fasting serum C-peptide value > or =0.33 mmol/l, OR: 3.01 (95% CI 1.07-8.55, p = 0.03) and compliance with life-style interventions OR 12.66 (95% CI 3.73-51.57, p = 0.0001). CONCLUSION: Among adult patients with DKA, significant predictors of good glycaemic control are preserved beta-cell function and compliance with life style modifications. PMID- 15811147 TI - Low-fat, high-carbohydrate (low-glycaemic index) diet induces weight loss and preserves lean body mass in obese healthy subjects: results of a 24-week study. AB - BACKGROUND: The traditional treatment for obesity which is based on a reduced caloric diet has only been partially successful. Contributing factors are not only a poor long-term dietary adherence but also a significant loss of lean body mass and subsequent reduction in energy expenditure. Both low-fat, high carbohydrate diets and diets using low-glycaemic index (GI) foods are capable of inducing modest weight loss without specific caloric restriction. The purpose of this study was to investigate the feasibility and medium-term effect of a low-fat diet with high (low GI) carbohydrates on weight loss, body composition changes and dietary compliance. METHODS: Obese patients were recruited from two obesity outpatient clinics. Subjects were given advise by a dietician, then they attended biweekly for 1-hour group meetings. Bodyweight and body composition were measured at baseline and after 24 weeks. RESULTS: One hundred and nine (91%) patients completed the study; after 24 weeks the average weight loss was 8.9 kg (98.6 vs. 89.7 kg; p < or = 0.0001). There was a significant 15% decrease in fat mass (42.5 vs. 36.4 kg; p < or = 0.0001) and a decrease in lean body mass of 5% (56.1 vs. 53.3 kg; p < or = 0.0001). DISCUSSION: In this 6-month study, a low-fat, low-GI diet led to a significant reduction of fat mass; adherence to the diet was very good. Our results suggest that such a diet is feasible and should be evaluated in randomized controlled trials. PMID- 15811148 TI - Lipoic acid prevents collagen abnormalities in tail tendon of high-fructose-fed rats. AB - BACKGROUND: Non-enzymatic glycation of proteins, leading to chemical modification and cross-linking are of importance in the pathology of diabetic complications. We studied the effect of alpha-lipoic acid (LA) on the glycation and cross linking of collagen from tail tendon of high-fructose-fed rats. METHODS: The rats were divided into four groups of six each. Two groups of rats were fed with a high-fructose diet (60/100 g diet) and administered with either LA (35 mg/kg body weight, intraperitoneally) (FRU + LA) or saline (FRU) for 45 days. The other two groups were fed control diet containing starch (60/100 g diet) and administered with either saline (CON) or LA (CON + LA). The rats were maintained for 45 days and then sacrificed. Collagen was isolated from tail tendon and the extent of glycation, fluorescence, aldehyde and peroxidation levels were measured. The tail tendons were separated, and shrinkage temperature was also measured. Acid-soluble collagen was extracted from tail tendon and subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis. RESULTS: Fructose administration caused accumulation of collagen in tail tendon. Glycation, collagen-linked fluorescence, aldehyde and peroxide contents were elevated. The gel pattern of acid-soluble collagen from the fructose-fed rat showed elevated beta-component. These changes were alleviated by the simultaneous administration of LA. CONCLUSION: Administration of LA has a positive influence on tail tendon collagen glycation and other variables in high-fructose-fed rats. PMID- 15811150 TI - Patients treated by dental students in outreach: the first year of a pilot project. AB - This paper describes the patients treated by 4th year undergraduate students during the first year of a pilot outreach course to teach Restorative Dentistry in community clinics in 2001-02. Data were collected from 908 summaries of patient treatment completed by the students, and from 139 patient questionnaires. Some 75% of patients were aged between 16 and 64, 58% were female, and 16% had dental phobia or anxiety. Most lived locally to the clinic and 41% made their initial contact as an emergency or drop-in. Some 37% made only a single visit (including children treated as emergencies) but 22% made six or more visits. Did not attend (DNA) was a problem and 18% of patients DNA to complete their treatment. Students undertook the full range of restorative procedures, with the emphasis on direct restorations, preventive treatment and advice, scaling, extractions and emergency treatment. Patients' main reasons for attending the clinic were lay recommendation, the need for treatment, convenience, free treatment, or the lack of access to a dentist. Some 30% said they did not have or did not know of an alternative source of dental care, and half had not seen a dentist for at least 2 years. The study demonstrates that despite difficulties related to attendance, a suitable patient base can be established offering students the opportunity to provide comprehensive care for adults in a primary care setting. PMID- 15811151 TI - Computer-mediated instructional video: a randomised controlled trial comparing a sequential and a segmented instructional video in surgical hand wash. AB - BACKGROUND: Video-based instructions for clinical procedures have been used frequently during the preceding decades. AIM: To investigate in a randomised controlled trial the learning effectiveness of fragmented videos vs. the complete sequential video and to analyse the attitudes of the user towards video as a learning aid. MATERIALS AND METHODS: An instructional video on surgical hand wash was produced. The video was available in two different forms in two separate web pages: one as a sequential video and one fragmented into eight short clips. Twenty-eight dental students in the second semester were randomised into an experimental (n = 15) and a control group (n = 13). The experimental group used the fragmented form of the video and the control group watched the complete one. The use of the videos was logged and the students were video taped whilst undertaking a test hand wash. The videos were analysed systematically and blindly by two independent clinicians. The students also performed a written test concerning learning outcome from the videos as well as they answered an attitude questionnaire. RESULTS: The students in the experimental group watched the video significantly longer than the control group. There were no significant differences between the groups with regard to the ratings and scores when performing the hand wash. The experimental group had significantly better results in the written test compared with those of the control group. There was no significant difference between the groups with regard to attitudes towards the use of video for learning, as measured by the Visual Analogue Scales. Most students in both groups expressed satisfaction with the use of video for learning. CONCLUSION: The students demonstrated positive attitudes and acceptable learning outcome from viewing CAL videos as a part of their pre-clinical training. Videos that are part of computer-based learning settings would ideally be presented to the students both as a segmented and as a whole video to give the students the option to choose the form of video which suits the individual student's learning style. PMID- 15811152 TI - Continuing professional development--global perspectives: synopsis of a workshop held during the International Association of Dental Research meeting in Gothenburg, Sweden, 2003. Part 1: access, funding and participation patterns. AB - There appears to have been little previous research interest in continuing professional development (CPD) of dentists and the oral health team. This paper presents data and information on the following aspects of CPD in 17 countries in Asia, Australasia, Europe and North America: availability of different types of CPD, its providers, data on uptake of CPD courses and activities, and funding of CPD. The results indicate that lectures and hands-on skills courses were held in all 17 countries but the use of the Internet to deliver CPD was by no means universal. CPD was funded from a variety of sources including universities, governments and commercial companies. However, the only universal source of funding for CPD was dentists themselves. Data on participation were available from only three countries. Research issues based on these results will be listed in a second paper. PMID- 15811153 TI - Continuing professional development--global perspectives: synopsis of a workshop held during the International Association of Dental Research meeting in Gothenburg, Sweden, 2003. Part 2: regulatory and accreditation systems and evidence for improving the performance of the dental team. AB - This paper is the second in a series of two that report on continuing professional development (CPD). Details of the informants and the methodologies used were reported in the first paper. This paper reports the data and information presented on the topics of regulatory and accreditation systems for CPD and evidence that CPD improves the performance of the oral health team. By June 2003, participation in CPD was mandatory in most of the states of the USA, all Canadian Provinces, the UK and Latvia and was likely to become mandatory in a number of other countries in the near future. A variety of accreditation systems were reported including collecting CPD points, which in some countries were weighted depending on the type of CPD activity, and re-certification examinations. Very few studies for the effectiveness of dental CPD were identified. However, in general it was concluded that there is little evidence for the effectiveness of CPD for the oral health team. The main recommendation from this study is that a systematic review of the effectiveness of CPD in improving the performance of the oral health team and patient based outcomes be undertaken. A range of other research questions was also identified including: how can CPD be best matched to clinicians' needs rather than demands? PMID- 15811154 TI - Geriatric dentistry education in European dental schools. AB - As the numbers of elderly adults continue to grow within European populations, the need for dental students to be trained in the management of geriatric patients becomes increasingly important. Many dental schools have developed training programmes in geriatric dentistry in response to the changing oral health needs of older adults. The purpose of this on-line survey was to identify the current status of geriatric dentistry education in European dental schools. A questionnaire relating to the teaching of geriatric dentistry was posted on the Internet, and 194 dental schools in 34 European countries were invited to participate. Data from completed questionnaires were submitted to the investigators via email from 82 schools in 27 countries (42% response rate). Thirty-six percent of schools offered a specific geriatric dentistry course that included didactic teaching or seminar groups, 21% taught geriatric dentistry by means of organised presentations in the curriculum, and 36% taught the subject by occasional lectures. 7% of schools did not teach geriatric dentistry at all. A clinical component to the geriatric dentistry curriculum was reported by 61% of schools and 18% reported operating a specific geriatric dentistry clinic within the school. Of those providing clinical geriatric dentistry training, it was provided within the school in 45% of cases, with a further 29% of schools providing training both within the school and at a remote location. Seven percent of schools operated a mobile dental clinic for treating geriatric patients. Twenty-eight percent of schools had a geriatric programme director or a chairman of a geriatric section and 39% indicated that they plan to extend the teaching of geriatric dentistry in the future. Geriatric dental education has clearly established itself in the curricula of European dental schools although the format of teaching the subject varies widely. It is of concern that geriatric dentistry was not taught at all in 7% of schools. No data are available concerning whether or not geriatric dentistry is taught in the 58% non-responding schools. PMID- 15811155 TI - Stress, burnout and health in the clinical period of dental education. AB - The study examined the extent of stress, burnout and health problems experienced by fourth and fifth year dental students from the three universities of Dresden, Freiburg and Bern. The objectives of the study were to: (i) identify frequent sources of stress and to report the prevalence rates of burnout and health problems in dental students, (ii) determine the rate of students suffering from severe burnout symptoms and (iii) identify stress factors related to the burnout symptoms of emotional exhaustion and depersonalization. A total of 161 dental students from Dresden, Freiburg and Bern participated in the study. They completed the Psychosocial Stress Inventory, the Maslach Burnout Inventory and the Health Survey Questionnaire. Frequent sources of stress were limitation of leisure time, examination anxiety and the transition stress that was related to the adaptation to the demands of the clinical phase of dental education. Few differences existed between the students of the fourth and the fifth study year. Study-related stress was lowest in Bern and considerably higher in Dresden. Differences of mean levels of burnout symptoms were found only for the burnout dimension of emotional exhaustion. Students from Dresden and Freiburg were more emotionally exhausted than students from Bern, students from Dresden also reported more health problems than students from Bern or Freiburg. Ten per cent of the dental students suffered from severe emotional exhaustion, 17% complained about a severe lack of accomplishment and 28% reported severe depersonalization symptoms. Forty-four per cent of the variance of emotional exhaustion was explained by study-related factors such as lack of leisure time, examination anxiety and transition stress. The only predictor of depersonalization was a lack of social integration, accounting for 3% of the variance. A lack of social integration may be an indicator of low social competence which may cause difficulties in dealing with patients adequately and therefore result in depersonalization. The results indicate a need to identify the group of students who may have insufficient social skills for dealing adequately with the patients, and to train them accordingly. PMID- 15811160 TI - Drug-drug interactions and adverse drug reactions: the bollards and flashing lights syndrome. PMID- 15811161 TI - Amantadine for traumatic brain injury: does it improve cognition and reduce agitation? AB - OBJECTIVE: To review the available literature pertaining to amantadine as therapy for improving cognition and reducing agitation following a non-penetrating traumatic brain injury (TBI). DATA SOURCES: Clinical literature was accessed through MEDLINE (from 1966 to February 2004) and bibliographic searches. Key search terms included 'amantadine', 'traumatic brain injury', 'cognition', and 'agitation'. DATA SYNTHESIS: Amantadine is primarily used for treatment and prophylaxis of influenza A. Its ability to improve mentation and motor function in patients with head injury remains questionable. An evaluation of five clinical trials, two case reports, and one case series is conducted focusing on the use of amantadine following TBI. Patients in clinical trials were assessed using a variety of neuropsychological tools aimed at, among other things, assessing cognition and agitation. Although individual patient results varied, the majority of patients studied showed improved neuropsychological test scores. Similar observations are noted in the case reports and case series. Improvement in cognition and reduced agitation seems to occur with post-injury amantadine therapy. CONCLUSIONS: Amantadine is a reasonable option for improving cognition and reducing agitation following a TBI but confirmatory evidence of the efficacy the drug is necessary. PMID- 15811162 TI - Comparison of Prilosec OTC (omeprazole magnesium 20.6 mg) to placebo for 14 days in the treatment of frequent heartburn. AB - PURPOSE: Consumer surveys have identified an over-the-counter (OTC) medication that provides complete and long-lasting relief of frequent heartburn as an unmet consumer need. The purpose of the two identical studies reported in this paper was to evaluate the safety and effectiveness of 10.3 and 20.6 mg omeprazole magnesium, referred to as Ome-Mg 10 and Ome-Mg 20, respectively (equivalent to 10 and 20 mg omeprazole) for the treatment of frequent heartburn administered as a novel 14-day OTC regimen. SUBJECTS AND METHODS: Subjects with frequent heartburn (heartburn two or more days per week) took Ome-Mg 10, Ome-Mg 20, or placebo for 14 consecutive mornings. Statistical analyses compared percentage of subjects with no heartburn 24 h after the first dose, after the last dose (day 14), and percentage of days that subjects were heartburn-free. Nocturnal heartburn and heartburn rated no more than mild were also assessed. RESULTS: Twenty-four hours following the first dose, nearly 50% of subjects receiving Ome-Mg 20 reported no heartburn, and more than 80% receiving Ome-Mg 20 had no more than mild heartburn. Both doses were significantly more effective than placebo on days 1 and 14 for percentage of subjects heartburn-free for 24 h (P < or = 0.003), and across all 14 days for percentage of heartburn-free days (P < 0.001). Ome-Mg 20 was significantly more effective than placebo in preventing nocturnal heartburn across all 14 days (P < 0.001). Ome-Mg was well tolerated. CONCLUSION: These trials demonstrated the safety and effectiveness of a novel 14-day regimen of Ome Mg 20 in completely preventing heartburn for 24 h establishing it as an excellent self-care treatment for frequent heartburn and supporting the approval of Prilosec OTC. PMID- 15811163 TI - Long-term tolerability of tramadol LP, a new once-daily formulation, in patients with osteoarthritis or low back pain. AB - INTRODUCTION: Tramadol hydrochloride is a centrally acting analgesic, which possesses opioid agonist properties and activates monoaminergic spinal inhibition of pain. An oral, once a day, sustained release formulation of tramadol is thought to be advantageous compared with immediate release preparations as it prevents plasma peaks associated with increased side-effects of the drug. It may also improve compliance. The purpose of the study was to assess the long-term safety of a new sustained-release formulation of tramadol (tramadol LP) in patients with knee or hip osteoarthritis and in patients with refractory low back pain. STUDY DESIGN: The design was a phase III, open, multicentre, international, tolerability study with tramadol LP at a dose titrated by the patient between 100 and 400 mg once daily, according to the intensity of pain. The treatment was administered for a continuous period of 4 weeks followed by an intermittent intake of 5 months in 204 patients. The safety criteria for evaluation were recording of adverse events, laboratory tests, electrocardiogram, radiography, global tolerability assessed by the patient and the investigators. RESULTS: Long term use of tramadol LP was reasonably well tolerated. Most of the reported adverse events were expected and occurred within the first month of treatment. Roughly half of the patients (49%) reported adverse events, of which 66% were related to treatment. Gastrointestinal events (nausea and vomiting) were the most frequent. Serious adverse events were reported in 6.4% of patients, from which only two cases were related to treatment. There was no sign of tolerance development and the percentage of patients presenting withdrawal symptoms after the end of treatment was low (6%). CONCLUSION: Long-term treatment with tramadol LP once daily is generally safe in patients with osteoarthritis or refractory low back pain. PMID- 15811164 TI - Dose-finding study of ibuprofen in patent ductus arteriosus using the continual reassessment method. AB - OBJECTIVE: Intravenous ibuprofen (IBU) has been found to be as effective as indomethacin for the treatment of patent ductus arteriosus (PDA) in preterm infants and has been associated with fewer adverse effects in comparative phase III studies. The dose regimen used (10-5-5 mg/kg/day) was based on limited pharmacokinetic data and no phase II study was available to determine the optimal dose of IBU for this indication. The present study was designed to determine the minimum effective dose regimen (MEDR) of IBU (one course) required to close ductus arteriosus in preterm infants. METHOD: A double-blind dose-finding study was conducted using the continual reassessment method, a Bayesian sequential design. Two distinct target closure rates were initially chosen according to postmenstrual age (PMA) at birth: 80% in infants with a PMA of 27-29 weeks, and 50% in infants with a PMA < 27 weeks. Forty neonates (20 in each PMA group) with PDA were treated between days 3 and 5 of life. Four different dose regimens were tested: loading doses of 5, 10, 15 or 20 mg/kg, followed by two doses (1/2 loading dose) at 24-h intervals. Efficacy was evaluated by echocardiography 24 h after the third infusion. RESULTS: In infants with a PMA of 27-29 weeks, the estimated MEDR was 10-5-5 mg/kg with a final estimated probability of success of 77% (95% credibility interval: 56-92%). The 15-7.5-7.5 mg/kg dose regimen had a better estimated probability of success (88%, 95% credibility interval: 68-97%), but resulted in more minor renal adverse effects. In contrast, in infants with a PMA < 27 weeks, the estimated MEDR was 20-10-10 mg/kg with an estimated probability of success of 54.8% (95% credibility interval: 22-84%), whereas the conventional dose regimen resulted in a low estimated probability of success (30.6%, 95% credibility interval: 13-56%). In these infants, compared with those with a PMA of 27-29 weeks, minor renal adverse effects were more frequent from the 10-5-5 mg/kg/day dose regimen and did not appear to be clearly dose related. CONCLUSION: This study confirms that the currently recommended dose regimen (10-5 5 mg/kg) of IBU is associated with a high closure rate (80%) and few adverse effects in premature infants with a PMA of 27-29 weeks. The failure rate was much higher below 27 weeks. A higher dose regimen (20-10-10 mg/kg) might achieve a higher closure rate. However, tolerability and safety of this dose regimen should be assessed in a larger population before considering the use of these doses for ductus arteriosus closure. PMID- 15811165 TI - Comparison of mirtazapine and fluoxetine in the treatment of major depressive disorder: a double-blind, randomized trial. AB - BACKGROUND: Depression is an international public health problem. The aim of this study was to compare the efficacy and tolerability of mirtazapine and fluoxetine treatment in a sample population consisting of Iranian patients suffering major depressive disorder. METHODS: Thirty-six inpatients and outpatients with a diagnosis of major depressive disorder (Diagnostic and Statistical Manual of Mental Disorders-IV) and a score > or = 18 on the 17-item Hamilton Rating Scale for Depression (HAM-D-17) were randomly assigned to 6 weeks of treatment with mirtazapine (30 mg/day) or fluoxetine (20 mg/day). Efficacy was assessed by HAM-D 17. Information about adverse events was obtained by questioning of participants and/or their examination. Assessments were performed at weeks 0, 1, 2, 3, 4 and 6. RESULTS: Sixteen of mirtazapine-treated patients and fifteen of fluoxetine treated patients completed the 6-week study period. Both treatment groups were well matched at baseline with respect to demographic and disease characteristics. Both drugs showed a significant improvement over the 6 weeks of treatment (P < 0.001). There was no statistically significant difference between the mean +/- SEM HAM-D scores of two groups at weeks 1, 2, 3, 4, and at the end point. There were no significant differences between two groups in terms of response to treatment (> or = 50% decrease from baseline in HAM-D-17 total score) and remission (HAM-D-17 score of < or = 7). None of the differences in reported adverse events was statistically significant. CONCLUSION: In this study, mirtazapine and fluoxetine were equally effective and well tolerated after 6 weeks of treatment in patients with major depressive disorder. PMID- 15811166 TI - Comparison of the effect of aminophylline and low PEEP vs. high PEEP on EGF concentration in critically ill patients with ALI/ARDS. AB - BACKGROUND: Acute respiratory distress syndrome (ARDS) remains a serious, often fatal, condition, despite progress in modern critical care treatment. Cytokines play important roles in the pathogenesis of the syndrome, although their roles in the evaluation and outcome have not been clearly elucidated yet. OBJECTIVES: We tested whether serum concentration of epidermal growth factor (EGF), as one of the important inflammatory mediators, changes with time and administration of mechanical ventilation and aminophylline. PATIENTS AND METHODS: Thirty patients [mean (SD): age = 56.6 (17.4) years] with ARDS were enrolled. After diagnosis based on inclusion and exclusion criteria, the patients were intubated and mechanically ventilated. Two hours after ventilation with definite positive end expiratory pressure (PEEP), aminophylline with a specific dose was started. Serum samples were obtained at five time points of 0, 2, 2.5, 4 and 8 h post-starting PEEP. RESULTS: Serum EGF concentration decreased after mechanical ventilation with PEEP (P < 0.05). The serum EGF concentrations 8 h after intervention was statistically lower in the low PEEP group than in the high PEEP group. The Acute Physiology and Chronic Health Evaluation (APACHE) Pi score and PaO2/FiO2 improved significantly after 8 h (P < 0.05). CONCLUSION: Beneficial effects of mechanical ventilation and aminophylline on APACHE Pi score and PaO2/FiO2 influence serum EGF levels. These findings may have relevance to the development of multisystem organ failure. PMID- 15811167 TI - Adoption of celecoxib and rofecoxib: a nationwide database study. AB - BACKGROUND AND OBJECTIVE: Cyclooxygenase 2-selective non-steroidal anti inflammatory drugs (NSAIDs, coxibs) are recommended primarily for patients at high risk of gastrointestinal bleeding, most of them being elderly. Our objective was to describe and analyse patient- and physician-related factors affecting the adoption of celecoxib and rofecoxib 2 years after their launch in Finland. METHODS: Retrospective analysis of the nationwide Prescription Register. Physicians who had issued at least 200 reimbursed prescriptions in 2002 (n = 12 033, 80% of working-age Finnish physicians) were involved in the analysis. RESULTS AND DISCUSSION: Excluding patients with rheumatoid arthritis (RA), almost one-fifth (18%) of NSAIDs prescriptions were for coxibs. In patients with RA the share was 25%. The share of coxib prescriptions of all NSAIDs increased with age of the patient. Over one half (58%) of coxib prescriptions were issued for patients under 65 years of age. Specialists in physical and rehabilitation medicine were the fastest adopters of coxibs: one-third of their NSAID prescriptions in 2002 were for coxibs. Primary care physicians were the most conservative both in adopting and favouring coxibs. CONCLUSIONS: Coxibs have gained the status of standard prescription NSAIDs within a few years. Their use should be restricted to patients who could benefit most from the use. Routine prescribing of expensive new drugs increases the drug bill without additional health gain. PMID- 15811168 TI - Grapefruit juice-nifedipine interaction: possible involvement of several mechanisms. AB - OBJECTIVE: To develop a model based on mean residence time for better understanding the effect of grapefruit juice on the metabolism of nifedipine (NIF). MATERIAL AND METHODS: Sixteen healthy volunteers from an urban population were included. For each trial, the subjects drank water, fresh grapefruit juice or bottled grapefruit juice. Thirty minutes later, the subjects took a 10 mg capsule of NIF, orally. Plasma concentration of NIF was measured and the kinetic parameters were calculated with a non-compartmental model. RESULTS: Grapefruit juice increased the bioavailability of NIF, but did not significantly reduce the drug's metabolism as shown by the approximately constant metabolite to parent drug AUC ratio (P = 0.948). There was no significant increase in the amount of non-metabolized drug absorbed during first-pass: 0.12 and 0.16 (P = 0.470) without and with grapefruit juices respectively. There was an increase in the relative bioavailability (P = 0.039) and the apparent volume of distribution (Vdm) (P = 0.025) of dehydronifedipine with grapefruit co-administration. A second peak was also observed in the NIF plasma-concentration profile when the drug is co-administered with grapefruit juice. Therefore, the most likely explanation for the double peak phenomenon is a delay in gastric emptying (+32 min with grapefruit juice) caused by the pH of grapefruit juice. CONCLUSION: This study shows that grapefruit juice interferes with the metabolism of NIF by inhibiting NIF metabolism and slowing down the rate of gastric emptying. This study also confirms that the metabolic inhibition is not a first pass effect, but is a secondary oxidative step. PMID- 15811169 TI - Population pharmacokinetic investigation of phenobarbital by mixed effect modelling using routine clinical pharmacokinetic data in Japanese neonates and infants. AB - The population pharmacokinetics of phenobarbital was evaluated using 69 serum concentration measurements obtained from the routine phenobarbital monitoring of 35 neonates and infants. The data were analysed using the nonlinear mixed effects model. A one-compartment open pharmacokinetic model with first-order elimination was used. Covariates screened were current bodyweight (TBW), gestational age, postnatal age (PNA), postconceptional age and gender. The final pharmacokinetic parameters were CL/F (mL/h) = 3.41.TBW (kg) + 1.64. PNA (weeks), Vd/F(L) = 1.09.TBW.(kg) [corrected] and F = 0.406 for oral administration and F = 1 for suppository. Application of the findings in this study to patient care may permit selection of an appropriate initial maintenance dosage to achieve target phenobarbital concentrations, thus enabling the clinician to achieve the desired therapeutic effect in neonates and infants. PMID- 15811170 TI - The impact of socioeconomic and demographic factors on the utilization of smoking cessation medications in patients hospitalized with cardiovascular disease in Nova Scotia, Canada. AB - OBJECTIVE: To determine whether any demographic or socioeconomic factors affect the use of smoking cessation medications in patients hospitalized with heart disease. METHOD: Data were obtained from the Improving Cardiovascular Outcomes in Nova Scotia (ICONS) Canada database, which includes a registry of all hospitalized patients with a diagnosis of ischaemic heart disease, congestive heart failure, or atrial fibrillation since October 1997. Patients agreeing to provide follow-up were sent an enrollment survey to determine demographic and socioeconomic factors including household income, educational background and private drug insurance plans. RESULTS: Between 15 October 1997 and 31 December 2000, 5442 patients who were current smokers and 270 patients using a smoking cessation medication were admitted to hospital registered in the ICONS database. An enrollment survey was completed by 1071 current smokers and 77 patients using a smoking cessation agent. CONCLUSION: Higher education level, presence of private drug insurance plans, and less difficulty paying for basic needs were associated with higher use of smoking cessation medications. PMID- 15811171 TI - High prevalence of orthostatic hypotension and its correlation with potentially causative medications among elderly veterans. AB - BACKGROUND: Orthostatic hypotension (OH) is defined as a reduction of systolic blood pressure of at least 20 mmHg, or diastolic blood pressure of at least 10 mmHg from a sitting to a standing position. It is a common physical finding among older adults and associated with significant morbidity and mortality. Use of medications that have the potential to induce OH, particularly concomitant use of several of such medications, is a major factor for the development of OH. OBJECTIVES: To describe the prevalence of symptomatic and asymptomatic OH in veterans aged 75 years and older attending a geriatric clinic, and to assess the association between OH and the number of potentially causative medications used. METHODS: Charts of all patients who attended a VA geriatric clinic (Michael E. DeBakey VA Medical Center) during the period of 1 June 2002 to 1 June 2003 were reviewed retrospectively for (i) the use of potentially causative medications, i.e. medications that were reported to cause OH in at least 1% of the general population and that were available in the VA formulary, (ii) the presence or absence of OH, and (iii) the presence or absence of symptomatic OH. Patients with primary autonomic dysfunction, Parkinson's disease, and patients who were unable to stand, or who had no assessment for both sitting and standing blood pressure for other reasons were excluded. RESULTS: A total of 505 individual patients attended the clinic during the study period, and 342 patients fit the inclusion criteria. About 189 of these patients (55%) had OH. Among patients with OH, 61 patients (33%) were symptomatic, including 52 patients who had falls. The prevalence of OH in patients receiving zero, one, two, and three or more potentially causative medications was 35, 58, 60 and 65% respectively. Receiving hydrochlorothiazide was associated with the highest prevalence of OH (65%), followed by receiving lisinopril (60%), trazodone (58%), furosemide (56%) and terazosin (54%). CONCLUSION: The prevalence of OH is very high in older veterans and significantly related to the number of concurrent causative medications used. Providers should be educated to reduce the amount of potentially causative medications in the elderly and better assess patients in which use of such medications is necessary to avoid symptomatic OH. PMID- 15811172 TI - Differences in the single-oral-dose pharmacokinetics and urinary excretion of paracetamol and its conjugates between Hong Kong Chinese and Caucasian subjects. AB - BACKGROUND AND OBJECTIVES: The present study was conducted to determine if ethnic differences exist for single oral dose pharmacokinetics of paracetamol and its conjugates between Hong Kong Chinese and Caucasian subjects. METHODS: Twenty healthy Chinese (n = 11) and Caucasian (n = 9) subjects, aged 21-44 years, 11 male and nine female, were given oral paracetamol syrup 20 mg/kg, following an overnight fast. Paracetamol and its metabolites (glucuronide, sulphate, cysteine and mercapturic acid conjugates) were measured in serial plasma samples (0.25, 0.5, 0.75, 1.0, 1.5, 2, 3,...,12, 24 h) and urine collections (0-24 h) by high performance liquid chromatography. RESULTS: In Chinese subjects, the (mean range) peak plasma concentration of paracetamol was 23.8 mug/mL (17.9-32.3) and time to attain this peak 0.66 h (0.5-0.75). This was lower (P < 0.015) at 18.7 microg/mL (14.4-22.9) and achieved later (P < 0.033) at 1.06 h (0.5-2.0) in Caucasians. In Chinese subjects, plasma levels of glucuronide were lower, sulphate higher and cysteine conjugates significantly lower than in Caucasians (P < 0.05). Chinese subjects excreted 6% more sulphate and 5% less glucuronide. They also excreted significantly less mercapturic acid conjugates (P < 0.001). DISCUSSION AND CONCLUSION: Chinese subjects show more rapid absorption of paracetamol, a tendency to produce less glucuronide but more sulphate conjugates and reduced production of cysteine and mercapturic acid conjugates. The latter may help to protect against hepatotoxicity following paracetamol overdose. PMID- 15811173 TI - A study of the in vitro interaction between lidocaine and premedications using human liver microsomes. AB - OBJECTIVE: To investigate potential interactions between lidocaine (lignocaine) metabolism and premedication drugs, i.e. psychotropic and antianxiety agents (diazepam, midazolam), hypnotics (pentobarbital, thiamylal), depolarizing neuromuscular blocking agents (vecuronium, pancuronium and suxamethonium), an antihypertensive agent (clonidine) and an H2-receptor blocking agent (cimetidine) using human liver microsomes in vitro. METHODS: The interaction effects between lidocaine and premedication were examined using human liver microsomal preparations and monitored for enzyme activity. The lidocaine and its main metabolite (monoethylglycinexylide) were measured by HPLC/UV. RESULTS: Lidocaine metabolism was non-competitively inhibited by midazolam (Ki = 77.6 microM). Thiamylal was a competitive inhibitor of lidocaine metabolism (Ki = 885 microM). Cimethidine, pancuronium and vecuronium weakly inhibited lidocaine metabolism in a concentration-depend manner over the therapeutic range in human liver microsomes. On the contrary, suxamethonium, pentobarbital and clonidine did not inhibit lidocaine metabolism over the therapeutic range in human liver microsomes. CONCLUSION: These results show that the interactions between lidocaine and midazolam and thiamylal are of potential toxicological and clinical significance. PMID- 15811174 TI - Clearance rates of cerivastatin metabolites in a patient with cerivastatin induced rhabdomyolysis. AB - We report on a patient who developed acute rhabdomyolysis after taking cerivastatin. A 74-year-old hypercholestrerolaemic woman taking cerivastatin (0.15 mg/day) for 22 days complained of general muscle weakness and muscle pain. Her serum creatinine phosphokinase level was 19,190 IU/L. Serum myoglobin was over 3000 ng/mL. Serum concentration of cerivastatin at 6 h after taking the last dose (0.15 mg) was 8062.5 ng/L, which was almost 5.7 times higher than that of normal persons. The serum concentration of cerivastatin showed that the half-life of cerivastatin in this patient was 22.4 h, compared with 2.4 h for normal controls. Cerivastatin is catabolized by cytochrome P450, 3A4 and 2C8 to M-1, and by 2C8 to M-23. The ratio of M-23 to M-1 in her serum was much lower than that of control persons (0.64 vs. 2.08). She had previously taken simvastatin which is metabolized by CYP3A4, without any sign and symptoms of rhabdomyolysis. These results suggest that the slowed clearance of cerivastatin in this patient might have been compounded by cytochrome P450, 2C8 dysfunction. PMID- 15811176 TI - Assessing sensitivity to change: choosing the appropriate change coefficient. AB - The past 20-years have seen the development and evaluation of many health status measures. Unlike the high standards demanded of those who conduct and report clinical intervention trials, the methodological rigor for studies examining the sensitivity to change of health status measures are less demanding. It is likely that the absence of a criterion standard for change in health status contributes to this shortcoming. To increase confidence in the results of these types of studies investigators have often calculated multiple change coefficients for the same patient sample. The purpose of this report is to identify the conflict that arises when multiple change coefficients are applied to the same patient sample. Three families of change coefficients based on different assumptions concerning the sample composition are identified: (1) the sample is homogeneous with respect to change; (2) subgroups of patients who truly change by different amounts exist; (3) individual patients, many of whom truly change by different amounts exist. We present several analyses which illustrate a major conceptual conflict: the signal (a measure's true ability to detect change) for some of these coefficients appears in the noise term (measurement error) of the others. We speculate that this dilemma occurs as a result of insufficient preparatory work such as pilot studies to establish the likely change characteristic of the patient population of interest. Uncertainty in the choice of change coefficient could be overcome by conducting pilot studies to ascertain the likely change characteristic of the population of interest. Once the population's change characteristic is identified, the choice of change coefficient should be clear. PMID- 15811177 TI - Inactivation of MAP kinase signalling in Myc transformed cells and rescue by LiCl inhibition of GSK3. AB - c-Myc oncogene is an important regulator of cell cycle and apoptosis, and its dysregulated expression is associated with many malignancies. Myc is instrumental in directly or indirectly regulating the progression through the G1 phase and G1/S transition, and transformation by Myc results in perturbed cell cycle. Also contributory to the control of G1 is the Ras effector pathway Raf/MEK/ERK MAP kinase. Together with GSK3, ERK plays an important role in the critical hierarchical phosphorylation of S62/T58 controlling Myc protein levels. Therefore, our main aim was to examine the levels of MAPK in Myc transformed cells in light of the roles of ERK in cell cycle and control of Myc protein levels. We found that active forms of ERK were barely detectable in v-Myc (MC29) transformed cells. Furthermore, we could only detect reduced levels of activated ERK in c-Myc transformed cells compared to the non-transformed primary chick embryo fibroblast cells. The addition of LiCl inhibited GSK3 and successfully restored the levels of ERK in v-Myc and c-Myc transformed cells to those found in non-transformed cells. In addition, LiCl stabilised Myc protein in the non transformed and c-Myc transformed cells but not in v-Myc transformed cells. These results can provide an important insight into the role of MAPK in the mechanism of Myc induced transformation and carcinogenesis. PMID- 15811178 TI - Mid-term Body Mass Index increase among obese and non-obese individuals in middle life and deprivation status: a cohort study. AB - BACKGROUND: In the UK, obesity is associated with a clear socioeconomic gradient, with individuals of lower socioeconomic status being more likely to be obese. Several previous studies, using individual measures of soecioeconomic status, have shown a more rapid increase in Body Mass Index (BMI) over time among adults of lower socioeconomic status. We conducted a study to further examine whether ecologically defined deprivation status influences within-individual BMI change during middle life, as the answer to this question can help determine optimal preventive strategies both for obesity per se, and its' associated socioeconomic disparities. METHODS: Anonymised records of participants to the Stockport population-based cardiovascular disease risk factor screening programme were analysed. Individuals aged 35-55 who had a first screening episode between 1989 and 1993, and a subsequent screening episode were included in the study. Deprivation status was defined using quintiles of the Townsend score. Mean annual BMI change by deprivation group was calculated using linear regression. Subsequently, deprivation group was included in the model as an ordinal variable, to test for trend. The modelling was repeated separately for individuals who were obese (BMI < 30) and non-obese at the time of first screening. In supplementary analysis, regression models were also adjusted for baseline BMI. RESULTS: Of 21,976 women and 19,158 men initially screened, final analysis included just over half of all individuals [11,158 (50.8%) women and 9,831 (51.3%) men], due to the combined effect of loss to follow-up and incomplete BMI ascertainment. In both sexes BMI increased by 0.19 kg/m2 annually (95% Confidence Intervals 0.15-0.24 for women and 0.16-0.23 for men). All deprivation groups had similar mean annual change, and there was no evidence of a significant deprivation trend (p = 0.801, women and 0.892, men). Restricting the analysis to individuals who were non-obese at baseline did not alter the results in relation to the lack of a deprivation effect. When restricting the analysis to individuals who were obese at baseline however, the findings were suggestive of an association of BMI increase with higher deprivation group, which was further supported by a significant association when adjusting for baseline BMI. CONCLUSION: In the study setting, the BMI of non-obese individuals aged 35-55 was increasing over time independently of deprivation status; among obese individuals a positive association with higher deprivation was found. The findings support that socioeconomic differences in mean BMI and obesity status are principally attained prior to 35 years of age. Efforts to tackle inequalities in mean BMI and obesity status should principally concentrate in earlier life periods, although there may still be scope for focusing inequality reduction efforts on obese individuals even in middle life. PMID- 15811179 TI - You might as well smoke; the misleading and harmful public message about smokeless tobacco. AB - BACKGROUND: Compared to smoking cigarettes, use of Western smokeless tobacco (ST) products is associated with a very small risk of life-threatening disease (with estimates in the range of a few percent of the risk from smoking, or even less). This means that smokers can realize substantial health benefits by switching to ST, an obvious substitute. But consumers and policy makers have little chance of learning that ST is much less dangerous than smoking because popular information provided by experts and advocates overstates the health risks from ST relative to cigarettes. METHODS: To examine the extent of this overstatement in one medium, we conducted a systematic review of websites containing information about ST and health risks. We examined the content of 316 relevant websites identified by a Google search. RESULTS: We found that when any substantive information about the risk from ST is given, the risk is almost universally conflated with the risk from cigarettes. Accurate comparative risk information was quite rare, provided by only a handful of websites, all appearing low in our search results (i.e., of low popularity and thus unlikely to be found by someone searching for information). About 1/3 of the websites, including various authoritative entities, explicitly claimed that ST is as bad as or worse than cigarettes. Most of the other sites made statements that imply the risks are comparable. CONCLUSION: Through these websites, and presumably other information provided by the same government, advocacy, and educational organizations, ST users are told, in effect, that they might as well switch to smoking if they like it a bit more. Smokers and policy makers are told there is no potential for harm reduction. These messages are clearly false and likely harmful, representing violations of ethical standards. PMID- 15811180 TI - Is post-trabeculectomy hypotony a risk factor for subsequent failure? A case control study. AB - BACKGROUND: Ocular hypotony results in an increased break down of the blood aqueous barrier and an increase in inflammatory mediator release. We postulate that this release may lead to an increased risk of trabeculectomy failure through increased bleb scarring. This study was designed to try to address the question if hypotony within one month of trabeculectomy for Primary Open Angle Glaucoma (POAG), is a risk factor for future failure of the filter. METHODS: We performed a retrospective, case notes review, of patients who underwent trabeculectomy for POAG between Jan 1995 and Jan 1996 at our hospital. We identified those with postoperative hypotony within 1 month of surgery. Hypotony was defined as an intraocular pressure (IOP) < 8 mmHg or an IOP of less than 10 mmHg with choroidal detachment or a shallow anterior chamber. We compared the survival times of the surgery in this group with a control group (who did not suffer hypotony as described above), over a 5 year period. Failure of trabeculectomy was defined as IOP > 21 mmHg, or commencement of topical antihypertensives or repeat surgery. RESULTS: 97 cases matched our inclusion criteria, of these 38 (39%) experienced hypotony within 1 month of surgery. We compared the survival times in those patients who developed hypotony with those who did not using the log-rank test. This data provided evidence of a difference (P = 0.0492) with patients in the hypotony group failing more rapidly than the control group. CONCLUSION: Early post-trabeculectomy hypotony (within 1 month) is associated with reduced survival time of blebs. PMID- 15811181 TI - An exceptional Albanian family with seven children presenting with dysmorphic features and mental retardation: maternal phenylketonuria. AB - BACKGROUND: Phenylketonuria is an inborn error of amino acid metabolism which can cause severe damage to the patient or, in the case of maternal phenylketonuria, to the foetus. The maternal phenylketonuria syndrome is caused by high blood phenylalanine concentrations during pregnancy and presents with serious foetal anomalies, especially congenital heart disease, microcephaly and mental retardation. CASE PRESENTATION: We report on an affected Albanian woman and her seven children. The mother is affected by phenylketonuria and is a compound heterozygote for two pathogenetic mutations, L48S and P281L. The diagnosis was only made in the context of her children, all of whom have at least one severe organic malformation. The first child, 17 years old, has a double-chambered right ventricle, vertebral malformations and epilepsy. She is also mentally retarded, microcephalic, exhibits facial dysmorphies and small stature. The second child, a girl 15 years of age, has severe mental retardation with microcephaly, small stature and various dysmorphic features. The next sibling, a boy, died of tetralogy of Fallot at the age of three months. He also had multiple vertebral and rib malformations. The subsequent girl, now eleven years old, has mental retardation, microcephaly and epilepsy along with facial dysmorphy, partial deafness and short stature. The eight-year-old child is slightly mentally retarded and microcephalic. A five-year-old boy was a premature, dystrophic baby and exhibits mental retardation, dysmorphic facial features, brachydactyly and clinodactyly of the fifth finger on both hands. Following a miscarriage, our index case, the youngest child at two years of age, is microcephalic and mentally retarded and shows minor facial anomalies. All children exhibit features of phenylalanine embryopathy caused by maternal phenylketonuria because the mother had not been diagnosed earlier and, therefore, never received any diet. CONCLUSION: This is the largest family suffering from maternal phenylketonuria reported in the literature. Maternal phenylketonuria remains a challenge, especially in woman from countries without a neonatal screening program. Therefore, it is mandatory to be alert for the possibility of maternal phenylketonuria syndrome in case of a child with the clinical features described here to prevent foetal damage in subsequent siblings. PMID- 15811182 TI - Rapid tests and urine sampling techniques for the diagnosis of urinary tract infection (UTI) in children under five years: a systematic review. AB - BACKGROUND: Urinary tract infection (UTI) is one of the most common sources of infection in children under five. Prompt diagnosis and treatment is important to reduce the risk of renal scarring. Rapid, cost-effective, methods of UTI diagnosis are required as an alternative to culture. METHODS: We conducted a systematic review to determine the diagnostic accuracy of rapid tests for detecting UTI in children under five years of age. RESULTS: The evidence supports the use of dipstick positive for both leukocyte esterase and nitrite (pooled LR+ = 28.2, 95% CI: 17.3, 46.0) or microscopy positive for both pyuria and bacteriuria (pooled LR+ = 37.0, 95% CI: 11.0, 125.9) to rule in UTI. Similarly dipstick negative for both LE and nitrite (Pooled LR- = 0.20, 95% CI: 0.16, 0.26) or microscopy negative for both pyuria and bacteriuria (Pooled LR- = 0.11, 95% CI: 0.05, 0.23) can be used to rule out UTI. A test for glucose showed promise in potty-trained children. However, all studies were over 30 years old. Further evaluation of this test may be useful. CONCLUSION: Dipstick negative for both LE and nitrite or microscopic analysis negative for both pyuria and bacteriuria of a clean voided urine, bag, or nappy/pad specimen may reasonably be used to rule out UTI. These patients can then reasonably be excluded from further investigation, without the need for confirmatory culture. Similarly, combinations of positive tests could be used to rule in UTI, and trigger further investigation. PMID- 15811183 TI - Angiotensinogen M235T gene variants and its association with essential hypertension and plasma renin activity in Malaysian subjects: a case control study. AB - BACKGROUND: Essential hypertension is a major public health concern worldwide where its prevalence accounts for various cerebrovascular diseases. A common molecular variant of angiotensinogen (AGT), the precursor of potent vasoactive hormone angiotensin II, has been incriminated as a marker for genetic predisposition to essential hypertension in some ethnics. This case-control study was designed not only to determine the association of the AGT M235T gene variants with essential hypertension, but also its relationship to Plasma Renin Activity (PRA) in subjects attending the Health Clinic, Kuala Lumpur, Malaysia. METHODS: The study involved 188 subjects, 101 hypertensives and 87 normotensives. Consents were obtained from all the participated subjects. M235T gene variants were investigated using allele specific polymerase chain reaction and PRA was determined by radioimmunoassay. Hypertensinogenic factors such as dietary habits, physical activity, smoking and drinking habits were assessed using a pre-tested questionnaire. RESULTS: The genotype and allele distribution of the M235T variant differed significantly in hypertensives and normotensives (chi2 = 23.184, P < 0.001 and chi2 = 21.482, P < 0.001, respectively). The odds ratio for hypertension was 1.36 (95% confidence interval 1.03-1.80) for subjects with homozygous mutated allele TT of the M235T variant compared with other genotypes or 1.98 (95% confidence interval 1.46-2.67) for those carrying T allele compared to those carrying M allele. Plasma Renin Activity is also significantly higher in hypertensive subjects (PRA = 3.8 +/- 2.5 ngAI/ml/hr for hypertensives, PRA = 2.6 +/- 1.3 ngAI/ml/hr for normotensives, P < 0.001), but was not significantly different between groups of genotypes (P = 0.118). CONCLUSION: The M235T variant of the AGT is significantly associated with essential hypertension whereas the genotype TT or allele T is a possible genetic marker or risk factor for hypertension in Malaysian subjects. PMID- 15811184 TI - Effect of a chemical manufacturing plant on community cancer rates. AB - BACKGROUND: We conducted a retrospective study to determine if potential past exposure to dioxin had resulted in increased incidence of cancer in people living near a former manufacturing plant in New South Wales, Australia. During operation, from 1928 to 1970, by-products of the manufacturing process, including dioxin and other chemical waste, were dumped into wetlands and mangroves, discharged into a nearby bay and used to reclaim land along the foreshore, leaving a legacy of significant dioxin contamination. METHODS: We selected 20 Census Collector Districts within 1.5 kilometres of the former manufacturing plant as the study area. We obtained data on all cases of cancer and deaths from cancer in New South Wales from 1972 to 2001. We also compared rates for some cancer types that have been associated with dioxin exposure. Based on a person's residential address at time of cancer diagnosis, or at time of death due to cancer, various geo-coding software and processes were used to determine which collector district the case or death should be attributed to. Age and sex specific population data were used to calculate standardised incidence ratios and standardised mortality ratios, to compare the study area to two comparison areas, using indirect standardisation. RESULTS: During the 30-year study period 1,106 cases of cancer and 524 deaths due to cancer were identified in the study area. This corresponds to an age-sex standardised rate of 3.2 cases per 1,000 person years exposed and 1.6 deaths per 1,000 person-years exposed. The study area had a lower rate of cancer and deaths from cancer than the comparison areas. The case incidence and mortality due to lung and bronchus carcinomas and haematopoietic cancers did not differ significantly from the comparison areas for the study period. There was no obvious geographical trend in ratios when comparing individual collector districts to New South Wales according to distance from the potential source of dioxin exposure. CONCLUSION: This investigation found no evidence that dioxin contamination from this site resulted in increased cancer rates in the potentially exposed population living around the former manufacturing plant. PMID- 15811185 TI - Genotyping DNA pools on microarrays: tackling the QTL problem of large samples and large numbers of SNPs. AB - BACKGROUND: Quantitative trait locus (QTL) theory predicts that genetic influence on complex traits involves multiple genes of small effect size. To detect QTL associations of small effect size, large samples and systematic screens of thousands of DNA markers are required. An efficient solution is to genotype case and control DNA pools using SNP microarrays. We demonstrate that this is practical using DNA pools of 100 individuals. RESULTS: Using standard microarray protocols for the Affymetrix GeneChip Mapping 10 K Array Xba 131, we show that relative allele signal (RAS) values provide a quantitative index of allele frequencies in pooled DNA that correlate 0.986 with allele frequencies for 104 SNPs that were genotyped individually for 100 individuals. The sensitivity of the assay was demonstrated empirically in a spiking experiment in which 15% and 20% of one individual's DNA was added to a DNA pool. CONCLUSION: We conclude that this approach, which we call SNP-MaP (SNP microarrays and pooling), is rapid, cost effective and promises to be a valuable initial screening method in the hunt for QTLs. PMID- 15811186 TI - A 2-year prospective study of patient-relevant outcomes in patients operated on for knee osteoarthritis with tibial osteotomy. AB - BACKGROUND: Tibial osteotomy is a treatment for younger and/or physically active patients suffering from uni-compartmental knee osteoarthritis. The open wedge osteotomy by the hemicallotasis technique includes the use of external fixation. The use of external fixation has several advantages, as early mobilization and the opportunity for optimal correction. However, the hemicallotasis technique has also been described as a cumbersome procedure for the patient. The aim of this study was to prospectively evaluate patient-relevant outcomes during the first 2 post-operative years. Especially the treatment period, during which external fixation was used, was closely monitored. METHODS: In an uncontrolled study, fifty-eight consecutive patients, 30 men and 28 women (mean age 54 years) were operated on by the hemicallotasis technique were evaluated with the patient relevant outcome measure Knee injury and Osteoarthritis Outcome Score (KOOS) preoperatively, during the treatment with external fixation, one week after removal of the external fixation, at 6 months, and at one and two years postoperatively. RESULTS: At the 2-year postoperative follow-up, all subscales of the KOOS were improved (p < 0.001), mostly in pain (41-80 on a 0-100 worst to best scale) and knee-related quality of life (21-61 on a 0-100 worst to best scale), compared to the preoperative status. Significant improvements in pain and other symptoms, function of daily life and quality of life were seen already during the treatment period (mean 98 +/- 18 days) with the external fixation. More demanding functions such as kneeling, squatting, jumping and running, were improved first after extraction of the external fixation device and the pins. CONCLUSION: Tibial osteotomy by the hemicallotasis technique yields large improvement in self-rated pain, function and quality of life, which persists over two years. Surprisingly, large improvements occurred already during the immediate post-operative period when the external fixation was still used. PMID- 15811187 TI - Breast conserving surgery versus mastectomy: cancer practice by general surgeons in Iran. AB - BACKGROUND: There appear to be geographical differences in decisions to perform mastectomy or breast conserving surgery for early-stage breast cancer. This study was carried out to evaluate general surgeons' preferences in breast cancer surgery and to assess the factors predicting cancer practice in Iran. METHODS: A structured questionnaire was mailed to 235 general surgeons chosen from the address list of the Iranian Medical Council. The questionnaire elicited information about the general surgeons' characteristics and about their work experience, posts they have held, number of breast cancer operations performed per year, preferences for mastectomy or breast conserving surgery, and the reasons for these preferences. RESULTS: In all, 83 surgeons returned the completed questionnaire. The results indicated that only 19% of the surgeons routinely performed breast conserving surgery (BCS) and this was significantly associated with their breast cancer case load (P < 0.01). There were no associations between BCS practice and the other variables studied. The most frequent reasons for not performing BCS were uncertainty about conservative therapy results (46%), uncertainty about the quality of available radiotherapy services (32%), and the probability of patients' non-compliance in radiotherapy (32%). CONCLUSION: The findings indicate that Iranian surgeons do not routinely perform BCS as the first and the best treatment modality. Further research is recommended to evaluate patients' outcomes after BCS treatment in Iran, with regard to available radiotherapy facilities and cultural factors (patients' compliance). PMID- 15811188 TI - Effects of glaucoma drugs on ocular hemodynamics in normal tension glaucoma: a randomized trial comparing bimatoprost and latanoprost with dorzolamide [ISRCTN18873428]. AB - BACKGROUND: Reduced choroidal perfusion is hypothesized to play a role in the pathogenesis of normal tension glaucoma. Thus the impact of antiglaucomatous eye drops on ocular perfusion has been the focus of recent research and the subject of intensive investigations. The present study investigates whether topically applied latanoprost or bimatoprost influence ocular perfusion in patients with normal tension glaucoma and compares these effects with that changes detected after the treatment with dorzolamide. METHODS: Ocular hemodynamics were assessed by color Doppler imaging (CDI) shortly before and after a one-month treatment with either latanoprost, bimatoprost or dorzolamide. Primary end-points of the study were peak systolic and end-diastolic blood flow velocities in the short posterior ciliary artery (SPCA) under the new therapy. Intraocular pressure (IOP) and additional perfusion parameters in the SPCA and other retrobulbar vessels were tracked as observational parameters. n = 42 patients with normal tension glaucoma were enrolled in the study. RESULTS: Systolic and diastolic blood flow velocities in the SPCA showed no significant alteration after the treatment with latanoprost or bimatoprost. Dorzolamide lead to increase of peak systolic velocity. IOP was reduced by all three agents in a range reported in the literature. CONCLUSION: Topically applied latanoprost and bimatoprost act in a hemodynamically neutral manner and have the capability to lower IOP even in patients with normal tension glaucoma and low initial IOP level. Dorzolamide accelerates blood flow in systole. None of the tested compounds has a negative impact on hemodynamics in the short posterior ciliary arteries. PMID- 15811189 TI - Attitudes to and management of fertility among primary health care physicians in Turkey: an epidemiological study. AB - BACKGROUND: The subject of infertility has taken its place in the health sector at the top level. Since primary health care services are insufficient, most people, especially women, keep on suffering from it all over the world, namely in underdeveloped or developing countries. The aim of this study was to determine primary care physicians' opinions about the approach to infertility cases and their place within primary health care services (PHCSs). METHODS: The study was conducted between October 2003 and April 2004. The study group comprised 748 physicians working in PHCSs. They were asked to fill in a questionnaire with questions pertaining to infertility support, laboratory and treatment algorithms, as well as the demographic characteristics. The data was evaluated using the chi square test, percentage rates and a logistic regression model. RESULTS: The multivariate analyses showed that having a previous interest in infertility and having worked for a postgraduate period of between 5-9 years and > or = 10 years were the variables that most positively influenced them in their approach to cases of infertility (p < 0.05, each one). Just 28.7% of the physicians indicated that they believed cases of infertility could be evaluated at the primary care level. The most frequently proposed reason for indicating 'difficulty in practice' (n = 533) was inadequate provision of equipment in PHCSs (55.7%). The physicians reported that they were able to perform most of the supportive treatments and proposals (between 64.6%-87.7%). The most requested laboratory investigations were the instruction of patients in taking basal body temperatures and semen analysis (89.7% and 88.7%, respectively). The most preferential course of treatment was that of sexually transmitted diseases (95.5%). CONCLUSION: It is clear that not enough importance is attached to the provision of care to infertile couples within PHCSs. This leads us to conclude that an integration of infertility services in primary care would be appropriate after strengthening the PHCSs. PMID- 15811190 TI - Parental exercise is associated with Australian children's extracurricular sports participation and cardiorespiratory fitness: A cross-sectional study. AB - BACKGROUND: The relationship between parental physical activity and children's physical activity and cardiorespiratory fitness has not been well studied in the Australian context. Given the increasing focus on physical activity and childhood obesity, it is important to understand correlates of children's physical activity. This study aimed to investigate whether parental exercise was associated with children's extracurricular sports participation and cardiorespiratory fitness. METHODS: The data were drawn from a nationally representative sample (n = 8,484) of 7-15 year old Australian schoolchildren, surveyed as part of the Australian Schools Health and Fitness Survey in 1985. A subset of 5,929 children aged 9-15 years reported their participation in extracurricular sports and their parents' exercise. Cardiorespiratory fitness was measured using the 1.6 km (1-mile) run/walk and in addition for children aged 9, 12 or 15 years, using a physical work capacity test (PWC170). RESULTS: While the magnitude of the differences were small, parental exercise was positively associated with children's extracurricular sports participation (p < 0.001), 1.6 km run/walk time (p < 0.001) and, in girls only, PWC170 (p = 0.013). In most instances, when only one parent was active, the sex of that parent was not an independent predictor of the child's extracurricular sports participation and cardiorespiratory fitness. CONCLUSION: Parental exercise may influence their children's participation in extracurricular sports and their cardiorespiratory fitness levels. Understanding the correlates of children's extracurricular sport participation is important for the targeting of health promotion and public health interventions, and may influence children's future health status. PMID- 15811192 TI - Australian public health policy in 2003 - 2004. AB - In Australia, compared with other developed countries the many and varied programs which comprise public health have continued to be funded poorly and unsystematically, particularly given the amount of publicly voiced political support.In 2003, the major public health policy developments in communicable disease control were in the fields of SARS, and vaccine funding, whilst the TGA was focused on the Pan Pharmaceutical crisis. Programs directed to health maintenance and healthy ageing were approved. The tertiary education sector was involved in the development of programs for training the public health workforce and new professional qualifications and competencies. The Abelson Report received support from overseas experts, providing a potential platform for calls to improve national funding for future Australian preventive programs; however, inconsistencies continued across all jurisdictions in their approaches to tackling national health priorities. Despite 2004 being an election year, public health policy was not visible, with the bulk of the public health funding available in the 2004/05 federal budget allocated to managing such emerging risks as avian flu. We conclude by suggesting several implications for the future. PMID- 15811191 TI - Anti-obesity effects of chikusetsusaponins isolated from Panax japonicus rhizomes. AB - BACKGROUND: The rhizomes of Panax japonicus are used as a folk medicine for treatment of life-style related diseases such as arteriosclerosis, hyperlipidemia, hypertension and non-insulin-dependent diabetes mellitus as a substitute for ginseng roots in China and Japan. Obesity is closely associated with life-style-related diseases. This study was performed to clarify whether chikusetsusaponins prevent obesity induced in mice by a high-fat diet for 9 weeks. METHODS: We performed two in vivo experiments. In one, female ICR mice were fed a high-fat diet with or without 1 or 3% chikusetsusaponins isolated from P. japonicus rhizomes for 9 weeks. In the other, lipid emulsion with or without chikusetsusaponins was administered orally to male Wistar rats, and then the plasma triacylglycerol level was measured 0.5 to 5 h after the orally administered lipid emulsion. For in vitro experiments, the inhibitory effects of total chikusetsusaponins and various purified chikusetsusaponins on pancreatic lipase activity were determined by measuring the rate of release of oleic acid from triolein in an assay system using triolein emulsified with lecithin. RESULTS: Total chikusetsusaponins prevented the increases in body weight and parametrial adipose tissue weight induced by a high-fat diet. Furthermore, consumption of a high-fat diet containing 1 or 3% total chikusetsusaponins significantly increased the fecal content and triacylglycerol level at day 3 compared with the high-fat diet groups. Total chikusetsusaponins inhibited the elevation of the plasma triacylglycerol level 2 h after the oral administration of the lipid emulsion. Total chikusetsusaponins, chikusetsusaponin III, 28 deglucosyl-chikusetsusaponin IV and 28-deglucosyl-chikusetsusaponin V inhibited the pancreatic lipase activity. CONCLUSION: The anti-obesity effects of chikusetsusaponins isolated from P. japonicus rhizomes in mice fed a high-fat diet may be partly mediated through delaying the intestinal absorption of dietary fat by inhibiting pancreatic lipase activity. The present study clearly indicated that the saponin fractions of P. japonicus rhizomes had a significant anti obesity action and supports the traditional usage as a substitute drug for ginseng roots. PMID- 15811193 TI - Introduction: the role of the thiazolidinediones in the cardiovascular risk management of type 2 diabetes. AB - SCOPE: This article serves as an introductory overview to this supplement which covers type 2 diabetes as an atherosclerotic disease, the evidence base for treatment of the various vascular risk factors, and provides a detailed appraisal of the potential for thiazolidinediones to play a major role in overall diabetes management, not just for glycaemia, but also from the point of view of cardio vascular disease. CONCLUSION: We are clearly entering into an extremely interesting time in the management of type 2 diabetes. The thiazolidinediones have the potential to target a fundamental defect in type 2 diabetes as well as to improve CV risk in this extremely high risk group of patients. Time will tell whether the obvious potential of these agents will result in dramatically improved clinical outcomes. PMID- 15811194 TI - Inflammatory, atherothrombotic aspects of type 2 diabetes. AB - BACKGROUND: There is a tight association between diabetes and cardiovascular disease - both occur more commonly together than independently. The development of vascular disease is dependent upon complex interactions between a number of metabolic pathways involving both fluid phase proteins and cellular components. Inflammation and coagulation are two intimately linked processes that are co regulated. The characteristic cluster of risk factors - hypertension, hyperinsulinaemia, hyperglycaemia and lipid abnormalities, which are associated with insulin resistance - have been expanded to include inflammation and thrombotic risk. Studies in patients with diabetes indicate a higher prevalence of both inflammatory cells and thrombosis in coronary plaques in comparison to non-diabetic subjects and measures of C-reactive protein predict the development of both diabetes and cardio vascular disease in prospective studies. SCOPE: This review (based on MEDLINE searches, 1990 to 2005) looks at the inflammatory, atherothrombotic aspects of type 2 diabetes that may be involved in the accelerated development of vascular disease in the population with diabetes. PMID- 15811195 TI - PPARgamma and atherosclerosis. AB - BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)gamma regulates a number of cellular processes that affect glucose homeostasis, endothelial function and vessel wall inflammation, as well as protecting against cardiovascular complications that occur in diabetes. Thiazolidinediones are PPARgamma agonists that are in clinical use for the treatment of type 2 diabetes. Accumulating evidence indicates that thiazolidinediones may exert cardioprotective effects at each stage of atherogenesis. SCOPE: This paper reviews preclinical and clinical evidence (identified from a search of MEDLINE databases) supporting a beneficial cardiovascular effect of thiazolidinediones and discusses the implications of these data for the optimal use of thiazolidinediones in clinical practice. FINDINGS: In vitro animal model and clinical studies indicate that thiazolidinediones correct endothelial dys function, suppress chronic inflammatory processes, reduce fatty streak formation, delay plaque evolution and vessel wall thickening and enhance plaque stabilization and regression. CONCLUSION: Thus, thiazolidinediones show potential as potent anti-inflammatory, antithrombotic agents that could both improve glucose levels and the long-term cardio vascular risk related to atherosclerosis in patients with type 2 diabetes. PMID- 15811196 TI - Diabetes and cardiovascular risk markers. AB - BACKGROUND: People with type 2 diabetes generally carry an array of risk factors for cardiovascular disease (CVD), including hyperglycaemia, dyslipidaemia, alterations in inflammatory mediators and coagulation/thrombolytic parameters, as well as other 'non-traditional' risk factors, many of which may be closely associated with insulin resistance. Consequently, rates of CVD mortality and morbidity are particularly high in this population. Targeting hyperglycaemia alone does not reduce the excess risk in diabetes, highlighting the need for aggressive treatment of other risk factors. SCOPE: This is a review of cardiovascular risk markers in diabetes, based on MEDLINE and EMBASE literature searches (1994-2004). FINDINGS: Although, the current use of statin therapy is effective at reducing low-density lipoprotein (LDL)-cholesterol, residual risk remains from other independent lipid and non-lipid factors. The peroxisome proliferator-activated receptor-gamma(PPARgamma) appears to be intimately involved in regulating risk markers at multiple levels. Ligands that activate PPARgamma, which include the thiazolidinedione (TZD) insulin-sensitizing agents used to manage type 2 diabetes, display a number of potential anti-atherogenic properties, including effects on high-density lipoprotein (HDL) cholesterol and triglycerides, as well as other beneficial non-lipid effects, such as regulating levels of mediators involved in inflammation and endothelial dysfunction. Data from several sources suggest that simple strategies combining TZDs and statins could have complementary effects on CVD risk factors profiles in diabetes, alongside the ability to control glycaemia. CONCLUSION: It is hoped that studies currently underway will provide insights into the value of such treatment approaches in terms of reducing the excess CVD risk, morbidity and mortality associated with type 2 diabetes. PMID- 15811197 TI - Diabetic dyslipidaemia: insights for optimizing patient management. AB - BACKGROUND: Lipid abnormalities in people with diabetes are likely to play an important role in the development of atherogenesis. These lipid disorders include potentially atherogenic quantitative (increased triglyceride levels and decreased high-density lipoprotein-cholesterol [HDL-C] levels) and qualitative abnormalities of lipoproteins (changes in lipoprotein size, increase in triglyceride content of low-density lipoprotein (LDL) and HDL, glycation of apoproteins and increased susceptibility of LDL to oxidation). Guidelines from the two main diabetes organizations, the International Diabetes Federation and the American Diabetes Association, recommend the aggressive management of diabetic dyslipidaemia to reduce the risk of cardiovascular disease (CVD). Statins are the first choice pharmacological therapy to address diabetic dyslipidaemia due to their effectiveness at lowering LDL-C levels in patients with diabetes. Fibrates (peroxisome proliferator-activated receptor [PPAR]alpha ligands) target another aspect of dyslipidaemia by lower ing triglycerides (to a greater extent than statins) and raising HDL-C levels, especially when baseline levels are low. The PPARgamma agonist, pioglitazone appears to affect lipid metabolism by decreasing plasma triglycerides, increasing HDL-C and decreasing the number of small, dense atherogenic LDL particles. SCOPE: This paper provides a review of the current literature (based on searches of MEDLINE and EMBASE from 1985 to 2005, inclusive) supporting the recommendations for the management of dyslipidaemia among patients with type 2 diabetes, including new strategies involving drug combinations that achieve good glycaemic and lipidaemic control that could potentially reduce the morbidity and mortality associated with type 2 diabetes. PMID- 15811198 TI - Outcome studies in type 2 diabetes. AB - BACKGROUND: Outcome studies are used to measure clinically meaningful primary end points, such as mortality and cardiovascular morbidity. However, few outcome trials have been conducted exclusively in people with diabetes; the majority of conventional diabetes trials use surrogate end points that may or may not translate into clinical benefits. Our current knowledge of the effects of pharmacotherapies on cardiovascular risk in patients with diabetes has been gained from subgroups included in large-scale studies. Several trials with lipid modifying, antiplatelet and/or antihypertensive therapy, for example the recent Collaborative AtoRvastatin Diabetes Study, have included sufficient numbers of patients with diabetes to indicate that effective management can reduce cardio vascular risk in this patient population. The United Kingdom Diabetes Study and the Diabetes Control and Complications Trial provide important, but inconclusive data on the impact of glucose-lowering therapy on the incidence of cardiovascular complications in diabetes. Thiazolidinediones have only become available during the past few years, thus their effects were not assessed in these landmark trials. Ongoing studies in diabetic populations at high risk for further macrovascular events, such as the PROspective pioglitAzone Clinical Trial In macroVascular Events, have been designed to assess the effect of thiazolidinediones on cardiovascular outcome in patients with diabetes and should help to reinforce the importance of broad-based treatment of the multiple metabolic risk factors for cardiovascular disease in people with diabetes. SCOPE: This paper (based upon MEDLINE and EMBASE literature searches in the year range 1990-2005) reviews what we have learned from outcome studies up to the end of 2004 and looks at what we hope to learn from ongoing studies. PMID- 15811199 TI - The effect of bosentan on matrix metalloproteinase-9 levels in patients with systemic sclerosis-induced pulmonary hypertension. AB - OBJECTIVE: Systemic sclerosis (SSc) is an autoimmune disease that can potentially involve all tissues and organs of the human body. Based on the extent of the disease and organ involvement, different subsets of patients and organ involvement, different subsets of patients have been identified and several classifications proposed have been identified and several classifications proposed aiming to better stratify affected patients. The occurrence aiming to better stratify affected patients. The occurrence of pulmonary arterial hypertension (PAH), characterized of pulmonary arterial hypertension (PAH), characterized by altered tissue remodelling of the entire vessel wall, is the most severe complication that influences prognosis and survival. The molecular basis underlying the vascular damage is not yet known, but a family of enzymes named matrix metalloproteinases (MMPs), with a proteolytic activity towards several extra-cellular matrix (ECM) components, is likely to be involved. Recently, a dual inhibitor of endothelin-1, bosentan, has been successfully evaluated in clinical trials in PAH patients. RESEARCH DESIGN AND METHOD: The aim of this study is to investigate the expression of MMP-2 and MMP-9 in the serum of different subsets of SSc patients, and in patients treated with bosentan. Thirty five Caucasian patients with SSc were enrolled in the study, 12 of whom were found to have isolated PAH assessed by Doppler echocardiography. Eight patients fully met the inclusion criteria and were eligible for therapy with bosentan given at the dosage of 62.5 mg twice daily for 4 weeks followed by 125 mg twice daily for 50 weeks(15). The remaining patients (4/12) initiated bosentan therapy for a few weeks and, therefore, were considered at the baseline level only. Serum samples were analysed by gelatine zymography. RESULTS: The results suggest that MMP-9 but not MMP-2 is differently expressed according to the degree of organ involvement. In particular, MMP-9 serum levels are significantly decreased in PAH with respect to other subsets of SSc patients. Moreover, in bosentan-treated patients, after 12 months of therapy MMP-9 significantly (p < 0.05) increased and correlated with an improved clinical outcome, as measured by the '6-minutes walking' test. CONCLUSIONS: This is the first time that MMP-9 serum levels are reported to be down-regulated in PAH patients and up-regulated following bosentan treatment. Whether MMP-9 has a pathogenetic role in the vascular damage observed in PAH patients or it is a marker of bosentan effectiveness is not yet known. However, MMP-9 may be an important molecule that needs further investigation in SSc patients to better define its role. PMID- 15811200 TI - Efficacy of ezetimibe in patients with statin-resistant and statin-intolerant familial hyperlipidaemias. AB - OBJECTIVE: To investigate the efficacy of the cholesterol absorption inhibitor ezetimibe in patients with refractory familial hyperlipidaemia or intolerant to statin therapy. METHODS: This prospective study assessed the safety and efficacy of ezetimibe in 200 patients with refractory familial hyperlipidaemias not achieving a low-density-lipoprotein (LDL) cholesterol < 3 cholesterol < 3 mmol/L (116 mg/dL) including 22% intolerant to all statin therapy, many consuming intolerant to all statin therapy, many consuming sterol-containing products. RESULTS: Ezetimibe monotherapy resulted in 7% and 11% reductions in LDL cholesterol and apolipoprotein B respectively. Ezetimibe-statin combination therapy reduced LDL-cholesterol by an additional 11 +/- 27% and apolipoprotein B by 11 (+79 to -18)%. There was a similar response between various sub-groups but a wide variation within groups with the greatest effect seen in patients groups with the greatest effect seen in patients under-responding to statins. The number of patients achieving the LDL-C target of 3 mmol/L rose from 5.5% to 18%. Non significant effects included a 5 (+78 to -470)% reduction in triglycerides, 8 +/- 36% increment in HDL-cholesterol, 21 (+35 to -82)% reduction in C-reactive protein and a 1 (+20 to -50)% increase in alanine transaminase. No effects were seen on creatinine, creatine kinase, or insulin resistance. Fourteen patients (7%) discontinued ezetimibe: seven due to gastrointestinal side-effects, one patient developed an ezetimibe-induced hypercholesterolaemia (x 1.5), one developed ezetimibe-induced hypertriglyceridaemia (x 7) and five discontinued for other reasons. CONCLUSION: Ezetimibe is a useful addition to statins in patients with familial hyperlipidaemias but shows a highly variable response profile. PMID- 15811201 TI - Do older patients feel able to cope with illness following telephone consultations? A multi-practice survey in the UK. AB - BACKGROUND: Worldwide, patients are offered a telephone consultation as a first point of access to a health care professional. Whilst older patients form a significant proportion of callers, this group has expressed reservation about consultations that preclude a physical examination. OBJECTIVE: This study compares how two groups of patients (< 70 years, > or = 70 years) perceive their ability to cope with illness following a telephone consultation. METHODS: Nine hundred and fifty-six patients from deprived inner city areas receiving telephone consultations with a General Practitioner (GP)/Family doctor or nurse at five practices in South Yorkshire, UK were surveyed within 24 h of the consultation. A 49% response rate was achieved; the mean age of respondents in the respective groups was 78.5 years and 48 years. The questionnaire included the Patient Enablement Instrument (PEI), a self-reported measure of patients' ability to cope with illness measure of patients' ability to cope with illness and supplementary questions on access to GPs in the practice. RESULTS: There was a low response rate from younger patients and males in particular. Ethnic minority groups were hardly represented among the respondents. How ever, scores on the PEI following telephone consultations were no different between older and younger respondents (Mann-Whitney U test, p = 0.56). Regression analysis with PEI scores as the dependent variable and fitting age and differences in responses to the questionnaire as explanatory variables yielded an R(2) of 0.02 suggesting that 'age' per se was a poor predictor of 'enablement'. CONCLUSIONS: These data support an inclusive policy in the provision of telephone access to health care professionals in primary care. We conclude that older patients are not disadvantaged by consulting a primary care professional by telephone. PMID- 15811202 TI - Efficacy, safety and tolerability of duloxetine 60 mg once daily in major depression. AB - BACKGROUND: Major depressive disorders (MDD) present a significant public health problem, in terms of burden for individual sufferers, their families and society as a whole. Recently, dualacting antidepressants, which block both serotonin (5 HT) and noradrenaline (NA) reuptake, have been developed with the hope of improving depression treatment outcomes. Duloxetine is a dual reuptake inhibitor of 5-HT and NA that has recently been licensed in the USA for the treatment of MDD. OBJECTIVE: This paper summarises efficacy and tolerability data for duloxetine with particular reference to the dose recommended for clinical use -- 60 mg once daily. Papers relating to duloxetine 60 mg once daily were identified through Medline searches and the publication databases at Eli Lilly/Boehringer Ingelheim. FINDINGS: Randomised, placebo-controlled studies have demonstrated the efficacy of duloxetine 60 mg once daily for the treatment of depression in the short and long term. Thus, duloxetine 60 mg once daily was superior to placebo in reducing once daily was superior to placebo in reducing MDD symptoms according to the primary efficacy MDD symptoms according to the primary efficacy measure -- the 17-item Hamilton Depression Rating Scale (HAMD(17)). Significantly greater improvements in subfactors of HAMD(17) and quality of life measures were also seen. In addition, duloxetine has been shown significantly to reduce the general aches and pains that frequently accompany MDD. A recent placebo-controlled study demonstrated that duloxetine improved cognition and depression measures in depressed elderly patients. Duloxetine appears to have an acceptable tolerance. The most frequently observed adverse events with duloxetine were nausea, dry mouth and somnolence. Importantly, duloxetine did not appear to have a clinically significant effect on blood pressure. CONCLUSION: In summary, duloxetine 60 mg once daily is effective for the treatment of core depressive symptoms, as well as general aches and pains associated with depression. PMID- 15811203 TI - CARDS on the table: should everybody with type 2 diabetes take a statin? AB - BACKGROUND: Vascular complications are a major cause of morbidity and mortality in patients with type 2 diabetes mellitus (T2DM). The recently published Collaborative Atorvastatin Diabetes Study (CARDS) showed that atorvastatin (10 mg, once daily vs. placebo) markedly reduces vascular events in this high-risk population. The participants (n = 2838) were fairly typical T2DM = 2838) patients without cardiovascular disease and with at least one other risk factor: hypertension, retinopathy, albuminuria or current smoking. In the treatment group, coronary events were reduced by 36% (p = 0.001) and stroke by 48% (p = 0.001). The trial was terminated two years early on ethical grounds. The number needed to treat (NNT) was 27 for four years to prevent one event. However, the benefit may have been greater since a proportion of the placebo group received statin therapy. The benefit from statin treatment was independent of sex, age, baseline lipid levels, was independent of sex, age, baseline lipid levels, systolic blood pressure, retinopathy, albuminuria, systolic blood pressure, retinopathy, albuminuria, smoking or HbA smoking or HbA(1c). The frequency of adverse events did not differ between the groups. These findings support those of other statin trials. SCOPE: CARDS does not comment on renal function. However, other trials suggest that statins preserve renal function in those with and without DM. We discuss the CARDS study in this context in this brief overview paper. CONCLUSIONS: the evidence shows that we need to control glucose to prevent microvascular complications, to lower cholesterol to prevent macrovascular disease and to lower blood pressure to prevent both. It may be that the benefit of statins extends beyond a threshold low-density lipoprotein cholesterol level in patients with T2DM. More trials are needed in this field. PMID- 15811204 TI - The effect of granisetron on in vitro metabolism of doxorubicin, irinotecan and etoposide. AB - OBJECTIVE: Doxorubicin, irinotecan and etoposide are all associated with the debilitating side-effects of nausea and vomiting, thereby necessitating concomitant antiemetic therapy. However, this may increase the potential for drug drug interactions by inhibition or induction of the cytochrome P450 enzymes. A study was undertaken to investigate the effects of the 5-HT(3) -receptor antagonist granisetron on the metabolism of doxorubicin, irinotecan and etoposide in vitro in human liver microsomal preparations. RESEARCH DESIGN AND METHODS: Doxorubicin, 20 microM, irinotecan, 10 microM, and etoposide, 50 microM, were incubated in the presence of granisetron, 0 nM, 20 nM, 200 nM and 2000 nM, in human liver microsomal preparations (250 microg). The levels of unchanged doxorubicin, irinotecan and etoposide in the incubation mixture were determined by high-performance liquid chromatography. Positive controls were ketoconazole, 20 microM, a potent inhibitor of CYP3A metabolism, for irinotecan and etoposide and quercitrin, 2 mM, a potent inhibitor of aldo-keto reductase, for doxorubicin. RESULTS: In the absence of granisetron, unchanged doxorubicin, irinotecan and etoposide levels decreased by 34.2 +/- 5.5%, 21.3 +/- 2.9% and 13.4 +/- 1.6% of control, respectively. Ketoconazole prevented the breakdown of both irinotecan and etoposide, while quercitrin inhibited the breakdown of doxorubicin. Granisetron had no effect on the rate of reduction of doxorubicin, irinotecan or etoposide. CONCLUSIONS: The results from this study suggest that granisetron neither inhibits nor induces the enzymes involved in the metabolism of doxorubicin, irinotecan or etoposide. Thus, granisetron can be used safely to treat nausea and vomiting induced by these agents with minimal risk of drug-drug interactions. PMID- 15811205 TI - Timolol LA: a double-masked, active-controlled, randomized, crossover, comfort, ocular safety, and systemic bioavailability study in healthy volunteers. AB - PURPOSE: A new formulation of timolol with sorbic acid, timolol-LA (TLA) (Istaloldagger), has been developed which increases its ocular bioavailability. In the present study, we desired to evaluate the ocular comfort and systemic bioavailability of TLA in healthy volunteers in comparison to standard timolol maleate ophthalmic solution (TIM). METHODS: This study was a randomized, double masked, active-controlled, crossover evaluation of 0.5% TLA and 0.5% TIM, bid, in 12 normal healthy volunteers. Visits were at Days 0, 1, 2, 4 and 8 in each period, and there was a minimum 7 day interperiod washout. RESULTS: At all three post-dosing evaluation times (Day 1: Peak, Day 8: Trough, and Day 8: Peak), the 95% confidence interval for the difference between TLA and TIM was not more than 0.37 ng/mL. After administration of TLA, there was a greater incidence of burning/stinging and tearing, but not foreign body sensation, relative to TIM. In general, most symptoms were mild in intensity, and no subject discontinued treatment due to ocular discomfort. Both treatments decreased IOP to a similar level. CONCLUSION: TLA was relatively comfortable, with a safety profile consistent with further clinical development, and, with bid dosing (exaggerated [2X] that anticipated for clinical use), had a systemic bioavailability similar to that of TIM 0.5%, bid. The incidence of burning and stinging was higher with TLA than with TIM, although reports were mild in severity and did not result in any patient discontin uations. Although the results are of interest, further evaluation in a larger trial may be warranted. PMID- 15811206 TI - Clinical and economic issues associated with switching between triptans in clinical practice. AB - Seven oral triptans are now generally available for the acute treatment of migraine, and physicians may sometimes feel under pressure to switch patients from one triptan to another for nonclinical reasons. This commentary article provides advice on what information should be taken into account by the physician before they consider switching one triptan for another. We review recommendations on switching triptans from international guidelines for migraine management and relate these to data from a recently published study on the economic implications of switching triptans in the UK. Controlled clinical studies reveal that most of the oral triptans have broadly similar efficacy profiles. Switching triptans can therefore only be recommended if the patient experiences problems such as lack of efficacy or intolerable side effects following repeated use of the initial triptan. The retrospective database study revealed that most patients who had their triptan switched were subsequently switched again during a 15 month review period, most usually back to their original triptan. Overall, switching a patient's triptan led to increased costs (analysed as costs of medication and the GP consultation) to the healthcare provider. These data indicate that patients should only be switched from one triptan for another for clinical reasons and not for perceived economic reasons, i.e. cost of the medication. PMID- 15811207 TI - Pain-free rates with zolmitriptan 2.5 mg ODT in the acute treatment of migraine: results of a large double-blind placebo- controlled trial. AB - CONTEXT: Zolmitriptan 2.5 mg orally disintegrating tablets (ODT) allow patients to take the medication without fluids, which is convenient and avoids the risk of fluid-induced exacerbation of nausea/vomiting. OBJECTIVE: To evaluate the efficacy and tolerability of zolmitriptan 2.5 mg ODT taken as soon as possible after onset of a migraine. DESIGN: Multicenter, double-blind, parallel-group, placebo-controlled two-attack trial. SETTING: Outpatient headache clinics in the US. PATIENTS: 608 patients were randomized; 566 patients treated at least 1 migraine and were included in the tolerability assessment (565 patients were included in the intent-to-treat population). INTERVENTION: Patients were randomized to either zolmitriptan 2.5 mg ODT or placebo. Patients treated up to 2 migraine attacks as soon as possible after the start of their migraine pain. MAIN OUTCOME MEASURE: Pain-free rates at 2 h. RESULTS: Zolmitriptan 2.5 mg ODT (n = 281) demonstrated a significant pain-free rate vs. placebo (n = 284) at 2 h (40% vs. 20%, p < 0.001), 1.5 h (25% vs. 15%, p < 0.001), and 1 h (13% vs. 8%, p = 0.004). Sustained pain-free rate was significantly higher than placebo (31% vs. 15%; p < 0.001). Return to normal activities favored zolmitriptan 2.5 mg ODT at 1 h (p = 0.004), 1.5 h (p < 0.001), and 2 h (p < 0.001). Adverse events associated with zolmitriptan 2.5 mg ODT were those commonly reported with the use of triptans. CONCLUSIONS: Zolmitriptan 2.5 mg ODT, taken as early as possible after onset of a migraine attack, is effective in the treatment of migraine, producing a significantly higher pain-free rate than placebo 2 h post-dose, and also at the earlier time points of 1 h and 1.5 h post-dose. PMID- 15811208 TI - Ibandronate produces significant, similar antifracture efficacy in North American and European women: new clinical findings from BONE. AB - OBJECTIVES: BONE (oral iBandronate Osteoporosis vertebral fracture trial in North America and Europe) determined whether less frequent dosing of ibandronate (dose free interval > 2 months) provided similar antifracture efficacy to daily dosing. As osteoporosis medications must be effective across different populations, an additional objective of BONE was to investigate and report the effect of oral ibandronate in North American and European women, as described here. PATIENTS AND METHODS: BONE was a randomized, double-blind, placebo-controlled, fractureprevention study in 2946 postmenopausal women (age 55 years-80 years; > or = 5 years since menopause) with osteoporosis (low lumbar spine bone mineral density and one to four prevalent vertebral fractures [T4-L4]). Participants received daily calcium (500 mg) and vitamin D (400 IU) plus either placebo, oral daily ibandronate (2.5 mg) or oral intermittent ibandronate (20 mg every other day for 12 doses every 3 months). The efficacy and tolerability of ibandronate were assessed independently in both North American and European populations. RESULTS: Consistent, significant efficacy was observed in the North American (new vertebral fracture risk reduction: 60% and 54% with daily and intermittent ibandronate, respectively) and European patient populations (50% and 48%, respectively). Both ibandronate regimens also significantly reduced the incidence of new, worsening, and acute clinical, vertebral fractures. Daily and intermittent ibandronate significantly increased bone density at the spine in both North American (5.4% and 4.4% vs. baseline with daily and intermittent ibandronate, respectively) and European (7.1% and 6.3% vs. baseline, respectively) populations. Significant increases were also observed for total hip bone density (2.6% and 3.7% vs. baseline for daily, and 2.5% and 3.1% for intermittent; North American and European populations, respectively). Comparable, significant decreases in biochemical markers of bone turnover (reductions in urinary excretion of C-telopeptide levels of 53.5% and 67.1% vs. baseline for daily, and 50.0% and 53.8% for intermittent; North American and European populations, respectively) were also observed in both populations (p < 0.004 for all cited measurements in each ibandronate group vs. placebo). Oral ibandronate was well tolerated in both North American and European patients, with a safety profile similar to placebo. CONCLUSIONS: Oral ibandronate, administered daily or intermittently, effectively reduced vertebral fracture risk in North American and European women with postmenopausal osteoporosis. These results demonstrate the efficacy of ibandronate administered with extended dose-free intervals, regardless of patients' geographical origin. Research investigating other less frequent ibandronate regimens, such as once-monthly oral administration, is underway. PMID- 15811209 TI - Use of arsenic trioxide in haematological malignancies: insight into the clinical development of a novel agent. AB - BACKGROUND: Arsenic trioxide delivers high rates of complete clinical remission in patients with relapsed/refractory acute promyelocytic leukaemia (APL), and is associated with high rates of molecular remission as indicated by PCR negativity for the PML-RARalpha gene. OBJECTIVE: Mitochondria are considered to be the primary intracellular target of arsenic trioxide, and preclinical and mechanistic studies suggest that this agent may have broad applicability in haematological and other malignancies. Investigations of this agent are ongoing in a range of haematological malignancies, and studies in newly diagnosed APL, acute myeloid leukaemia (AML), myelodysplastic syndromes (MDS), multiple myeloma (MM) and chronic myelogenous leukaemia (CML) are reviewed here using published articles and presentations at international congresses to June 2004. Medline was used to source published preclinical and clinical data, and abstract databases and publications from relevant major international haematology/oncology congresses were searched to source updates of preclinical and clinical trial data. FINDINGS: Accumulating data indicate that arsenic trioxide may be a useful addition to the therapeutic regimens that have been so successful in treating newly diagnosed APL, and investigations are ongoing to incorporate this agent into the first-line APL treatment paradigm. Preliminary data from clinical studies indicate that arsenic trioxide has clinical activity as a single agent in MDS and MM, and combination therapies are being investigated. In MM, the combination regimens under study incorporate ascorbic acid, which can enhance the efficacy of arsenic trioxide by reducing intracellular glutathione concentrations. In CML, arsenic trioxide is being investigated in combination with imatinib mesylate in patients who have failed initial imatinib treatment. In AML, although results with single agent arsenic trioxide were not encouraging, treatment using arsenic trioxide in combination with ascorbic acid is a proposed strategy in elderly patients not able to withstand intensive chemotherapy. CONCLUSION: This versatile agent has a predictable and manageable safety profile and avoids many of the severe toxicities associated with conventional chemotherapies. Ongoing clinical studies will help to define the role of arsenic trioxide in the treatment of haematological malignancies. PMID- 15811210 TI - How treatment priorities influence triptan preferences in clinical practice: perspectives of migraine sufferers, neurologists, and primary care physicians. AB - BACKGROUND: In treating migraine sufferers, physicians can choose from among seven triptans with different attributes. OBJECTIVE: To develop a system for selecting an oral triptan based on treatment priorities of migraine sufferers, neurologists, and primary care physicians (PCPs) in the United States, and evidence-based performance of triptans in clinical trials. METHODS: The TRIPSTAR project combines data on the treatment preferences of migraineurs and physicians with results from a meta-analysis of individual triptans, which evaluated their effectiveness on various clinical endpoints. Telephone interviews with migraine sufferers, neurol ogists, and PCPs were conducted to elicit individual views on the relative importance of a prespecified set of acute treatment outcomes. Four hundred and fifteen migraine sufferers, both triptan-experienced and triptan naive, were interviewed. Also, 200 board-certified neurologists and 200 PCPs provided information on migraine patients from their clinical practice. A multiattribute decision model for selecting an oral triptan was constructed using attribute importance weights collected at telephone interview and the meta analysis data, which were drawn from 53 clinical trials of 6 oral triptans. RESULTS: Efficacy attributes were rated significantly more important than tolerability or consistency in selecting an oral triptan, according to migraine sufferers and physicians. Freedom from cardiovascular adverse events was the most important tolerability attribute, according to migraine sufferers and physicians alike. Pain free at 1 h was the most important lower-level efficacy attribute for migraine sufferers, while sustained pain free was most important for physicians. When weighted treatment attributes were combined with meta-analysis data in a multi-attribute decision model, almotriptan 12.5 mg, eletriptan 80 mg, and rizatriptan 10 mg were significantly closer to the hypothetical ideal triptan than was suma triptan 100 mg. Triptans selected by the model were generally closer to the patient-specific ideal triptan than were the triptans prescribed by physicians. CONCLUSIONS: Almotriptan, eletriptan, and rizatriptan were the three triptans closest to the ideal, from the perspectives of migraine sufferers, PCPs, and neurologists alike. The TRIPSTAR model may be a potentially useful decision support tool to help physicians select the triptan most likely to produce a successful outcome in migraine sufferers. PMID- 15811211 TI - A randomized, investigator- masked clinical trial comparing the efficacy and safety of gatifloxacin 0.3% administered BID versus QID for the treatment BID versus QID for the treatment of acute bacterial conjunctivitis of acute bacterial conjunctivitis. AB - PURPOSE: To compare the efficacy and safety of To compare the efficacy and safety of gatifloxacin ophthalmic solution 0.3% (Zymar) administered BID versus QID in patients with acute bacterial conjunctivitis. METHODS: In a randomized, investigator-masked clinical trial, patients diagnosed with bacterial conjunctivitis (based on signs and symptoms) received gatifloxacin either BID or QID for 5 days. Visits were scheduled at day 0, day 3, and day 5. Conjunctival cultures were taken at each visit. The clinical cure rate at day 5 was determined for the entire patient population (primary endpoint). Additionally, clinical cure at day 5 was evaluated for a population of patients defined a priori (per protocol) as being culture positive at baseline and with no substantial protocol deviations. Safety was determined through recording of adverse events. Minimal inhibitory concentrations (MIC) and susceptibility of isolates to gatifloxacin were determined using a broth dilution method. RESULTS: Patient characteristics in both the BID and QID groups (N = 104) were similar in terms NN of baseline demographics and disposition. The clinical cure rate on day 5 in the entire, intentto-treat (ITT) population was 86.5% (45/52) in the gatifloxacin BID group and 71.2% (37/52) in the gatifloxacin QID group (95% CI: [-0.03, 30.80]; p = 0.096). In both treatment groups, 5/52 patients (9.6%) reported adverse events. The most common adverse event was conjunctivitis. No serious adverse events were reported. In the a priori-defined per-protocol (PP) population, the clinical cure rate on day 5 was 95.5% (21/22) in the gatifloxacin BID group and 85.7% (18/21) in the gatifloxacin QID group (95% CI: [-7.57, 21.05]; p = 0.294). At baseline, 96.1% (98/102) of the isolates were susceptible to gatifloxacin. The overall MIC(90) (mean +/- standard error of the mean) was 0.5 +/- 1.3 microg/mL. CONCLUSION: In this study, gatifloxacin 0.3% administered BID was as effective and as safe as gatifloxacin 0.3% administered QID for 5 days for the treatment of bacterial conjunctivitis. PMID- 15811212 TI - Safety, effectiveness, and duration of effect of BOTOX after switching from Dysport for blepharospasm, cervical dystonia, and hemifacial spasm dystonia, and hemifacial spasm. AB - BACKGROUND: Local injection of botulinum toxin type A is first-line treatment of blepharospasm, cervical dystonia, and hemifacial spasm; however, there is uncertainty about the optimal dose of toxin for each indication as well as dose conversion ratios that should be used when switching products in clinical practice. OBJECTIVE: The goal of this study was to compare the safety, effectiveness, and duration of clinical effect of BOTOX and Dysport and Dysportdagger after drug switching (Dysport to BOTOX) among patients with movement disorders. METHODS: A total of 48 patients diagnosed with blepharospasm (n = 27), cervical dystonia (n = 12), or hemifacial spasm (n = 9) were evaluated during a single-arm, crossover-design study in which each patient was his/her own historical control using a 5:1 or 4:1 conversion ratio of Dysport to BOTOX units. Patients were assessed (using Jankovic, Visual Analog Scale, general pain scale, Toronto Western Spasmodic Torticollis Rating Scale [TWSTRS] scores) after the last injection of Dysport and the first injection of BOTOX. Moreover, each patient kept a diary during treatment to track onset and duration of therapeutic effect. RESULTS: Although BOTOX and Dysport were both clinically effective, this effect was more significant with BOTOX compared to Dysport based on TWSTRS for cervical dystonia (p = 0.012), and Jankovic scores for blepharospasm (p = 0.006). Longer duration of effect also was noted with BOTOX than with Dysport (blepharospasm, 62.2 days vs 47.4 days (p = 0.001); cervical dystonia, 64.3 days vs 44.6 days (p = 0.014); hemifacial spasm, 65.1 days vs 41.8 days (p < 0.014), respectively). Of the 48 patients, 19 experienced at least one adverse drug reaction (ADR) during Dysport treatment, with the most commonly reported ADRs being ptosis for blepharospasm and hemifacial spasm and neck weakness for cervical dystonia. No patient reported an ADR during BOTOX treatment. CONCLUSIONS: Results suggest therapeutic effectiveness is enhanced with BOTOX compared to Dysport at a dosing ratio between 5:1 and 4:1 (Dysport:BOTOX). Safety and duration of therapeutic effect also are enhanced with BOTOX. Further research is needed. PMID- 15811213 TI - Predictors of bleeding risk and long-term mortality in patients with acute coronary syndromes. AB - OBJECTIVE: The aim of this review was to collate the published evidence on independent predictors of bleeding and late mortality in acute coronary syndrome (ACS) patients, to compare the two sets of risk factors, and to investigate whether bleeding is reported as a predictor of late mortality. RESEARCH DESIGN AND METHODS: Computerized searches, covering the period from 1999 to July 2004, were performed on MEDLINE, EMBASE, and the Cochrane Library database. Studies were eligible for inclusion in the review if they related to patients with ACS and included an assessment of risk factors for bleeding, mortality, or both. Studies that did not meet these criteria were excluded. MAIN OUTCOME MEASURES: A total of 937 studies were retrieved, of which 912 were excluded from the review because they did not meet the defined criteria. RESULTS: The available evidence suggests that any impact of bleeding on mortality is confined to the short term. In studies that showed a significant association between bleeding and risk of mortality, association between bleeding and risk of mortality, this was always related to in-hospital or 30-day mortality. By contrast, studies of long-term mortality consistently showed that bleeding was not an independent predictor. Furthermore, follow-up studies showed that adverse outcomes in hospital and within the first month were not related to 1-year mortality. This may reflect the beneficial impact of anticoagulant therapy on subsequent cardiovascular risk in patients with ACS, which outweighs any shortterm detrimental effect of bleeding. CONCLUSIONS: The available evidence suggests that any impact of bleeding on mortality in ACS patients appears to be confined to the short term, and long-term outcomes do not reflect the impact of in-hospital bleeding. PMID- 15811214 TI - Cost implications of post-surgical morbidity following blood transfusion in cancer patients undergoing elective colorectal resection: an evaluation in the US hospital setting. AB - OBJECTIVE: To estimate the cost implications of blood transfusions and related surgical site infections (SSIs) in cancer patients undergoing elective colorectal resection in the hospital setting in the United States (US). STUDY DESIGN: A modelling study was performed from the perspective of the hospital sector, based on published clinical outcomes from a study in Taiwan involving 2809 cancer patients who underwent elective colorectal resection using laparotomy and American treatment patterns. METHODS: Data on resource use were retrieved from published literature and from two American hospital centres specialising in colorectal cancer management. Decision analytical modelling was used to estimate the treatment costs and consequences of managing patients undergoing elective colorectal resection with and without blood transfusions. RESULTS: The expected treatment costs of managing patients who required and did not require a blood transfusion were estimated to be US dollars 19,869 (95% CI: 15 797; 23 150) and US dollars 14,586 (95% CI: 14 263; 14 886) per patient respectively. Expected treatment costs for those patients transfused with 1-3 units and > 3 units of blood were estimated to be US dollars 17,449 and US dollars 22,588 per patient respectively. CONCLUSION: This is one of the first studies to specifically address the cost implications of postsurgical morbidity following colorectal resection in cancer patients. The cost of managing cancer patients undergoing elective colorectal resection who require a blood transfusion is expected to be 36% more than that of non-transfused patients, largely resulting from the development of SSIs. PMID- 15811215 TI - A review of the metabolic effects of sibutramine. AB - BACKGROUND: Obesity is associated with an increased incidence of diabetes, hypertension, dyslipidaemia and coronary artery disease. Current management strategies of obesity include lifestyle management strategies of obesity include lifestyle interventions and pharmaco therapy. Sibutramine is a drug with established efficacy in weight reduction and maintenance of weight loss. It reduces food intake and attenuates the fall in reduces food intake and attenuates the fall in metabolic rate associated with weight loss. OBJECTIVE: To review the metabolic effects associated with sibutramine use. METHODS: Relevant articles were identified through a Medline search (up to December 2004). RESULTS: Weight loss with sibutramine treatment is associated with improved insulin sensitivity and a fall in glycosylated haemoglobin levels in type 2 diabetic patients. In most trials sibutramine exerted favourable effects on lipids, especially exerted favourable effects on lipids, especially on high density lipoprotein (HDL) cholesterol and triglycerides, as well as on the total:HDL cholesterol ratio. Sibutramine also lowers serum uric acid concentrations. Furthermore, this drug seems to favourably influence adipocytokines; it reduces serum leptin and resistin levels and increases adiponectin levels. Sibutramine also exerts a beneficial effect on hyper androgenaemia in obese women with polycystic ovary syndrome. Preliminary findings also suggest that weight loss following treatment with sibutramine is useful in patients with non-alcoholic fatty liver disease (NAFLD). CONCLUSION: Weight loss following sibutramine administration is associated with several favourable metabolic effects. PMID- 15811216 TI - Artificial life: more than just building and studying computational systems. PMID- 15811218 TI - The development of embodied cognition: six lessons from babies. AB - The embodiment hypothesis is the idea that intelligence emerges in the interaction of an agent with an environment and as a result of sensorimotor activity. We offer six lessons for developing embodied intelligent agents suggested by research in developmental psychology. We argue that starting as a baby grounded in a physical, social, and linguistic world is crucial to the development of the flexible and inventive intelligence that characterizes humankind. PMID- 15811219 TI - Learning from and about others: towards using imitation to bootstrap the social understanding of others by robots. AB - We want to build robots capable of rich social interactions with humans, including natural communication and cooperation. This work explores how imitation as a social learning and teaching process may be applied to building socially intelligent robots, and summarizes our progress toward building a robot capable of learning how to imitate facial expressions from simple imitative games played with a human, using biologically inspired mechanisms. It is possible for the robot to bootstrap from this imitative ability to infer the affective reaction of the human with whom it interacts and then use this affective assessment to guide its subsequent behavior. Our approach is heavily influenced by the ways human infants learn to communicate with their caregivers and come to understand the actions and expressive behavior of others in intentional and motivational terms. Specifically, our approach is guided by the hypothesis that imitative interactions between infant and caregiver, starting with facial mimicry, are a significant stepping-stone to developing appropriate social behavior, to predicting others' actions, and ultimately to understanding people as social beings. PMID- 15811220 TI - Brain-based devices for the study of nervous systems and the development of intelligent machines. AB - The simultaneous study of brain function at all levels of organization is difficult to undertake with current experimental tools. Present day electrophysiology only allows the recording of at most hundreds of neurons while an animal is performing a behavioral task. Because of this limitation and the sheer complexity of the nervous system, computational modeling has become essential in developing theories of brain function. Accordingly, our group has constructed a series of brain-based devices (BBDs), that is, physical devices with simulated nervous systems that guide behavior, to serve as a heuristic for testing theories of brain function. Unlike animal models, BBDs permit analysis of activity at all levels of the nervous system as the device behaves in its environment. Although the principal focus of developing BBDs has been to test theories of brain function, this type of modeling may also provide a basis for robotic design and practical applications. PMID- 15811221 TI - Evolutionary robotics: a new scientific tool for studying cognition. AB - We survey developments in artificial neural networks, in behavior-based robotics, and in evolutionary algorithms that set the stage for evolutionary robotics (ER) in the 1990s. We examine the motivations for using ER as a scientific tool for studying minimal models of cognition, with the advantage of being capable of generating integrated sensorimotor systems with minimal (or controllable) prejudices. These systems must act as a whole in close coupling with their environments, which is an essential aspect of real cognition that is often either bypassed or modeled poorly in other disciplines. We demonstrate with three example studies: homeostasis under visual inversion, the origins of learning, and the ontogenetic acquisition of entrainment. PMID- 15811222 TI - New robotics: design principles for intelligent systems. AB - New robotics is an approach to robotics that, in contrast to traditional robotics, employs ideas and principles from biology. While in the traditional approach there are generally accepted methods (e. g., from control theory), designing agents in the new robotics approach is still largely considered an art. In recent years, we have been developing a set of heuristics, or design principles, that on the one hand capture theoretical insights about intelligent (adaptive) behavior, and on the other provide guidance in actually designing and building systems. In this article we provide an overview of all the principles but focus on the principles of ecological balance, which concerns the relation between environment, morphology, materials, and control, and sensory-motor coordination, which concerns self-generated sensory stimulation as the agent interacts with the environment and which is a key to the development of high level intelligence. As we argue, artificial evolution together with morphogenesis is not only "nice to have" but is in fact a necessary tool for designing embodied agents. PMID- 15811223 TI - From wheels to wings with evolutionary spiking circuits. AB - We give an overview of the EPFL indoor flying project, whose goal is to evolve neural controllers for autonomous, adaptive, indoor micro-flyers. Indoor flight is still a challenge because it requires miniaturization, energy efficiency, and control of nonlinear flight dynamics. This ongoing project consists of developing a flying, vision-based micro-robot, a bio-inspired controller composed of adaptive spiking neurons directly mapped into digital microcontrollers, and a method to evolve such a neural controller without human intervention. This article describes the motivation and methodology used to reach our goal as well as the results of a number of preliminary experiments on vision-based wheeled and flying robots. PMID- 15811224 TI - Flexible couplings: diffusing neuromodulators and adaptive robotics. AB - Recent years have seen the discovery of freely diffusing gaseous neurotransmitters, such as nitric oxide (NO), in biological nervous systems. A type of artificial neural network (ANN) inspired by such gaseous signaling, the GasNet, has previously been shown to be more evolvable than traditional ANNs when used as an artificial nervous system in an evolutionary robotics setting, where evolvability means consistent speed to very good solutions--here, appropriate sensorimotor behavior-generating systems. We present two new versions of the GasNet, which take further inspiration from the properties of neuronal gaseous signaling. The plexus model is inspired by the extraordinary NO-producing cortical plexus structure of neural fibers and the properties of the diffusing NO signal it generates. The receptor model is inspired by the mediating action of eurotransmitter receptors. Both models are shown to significantly further improve evolvability. We describe a series of analyses suggesting that the reasons for the increase in evolvability are related to the flexible loose coupling of distinct signaling mechanisms, one "chemical" and one "electrical." PMID- 15811225 TI - Agency in natural and artificial systems. AB - We analyze the conditions for agency in natural and artificial systems. In the case of basic (natural) autonomous systems, self-construction and activity in the environment are two aspects of the same organization, the distinction between which is entirely conceptual: their sensorimotor activities are metabolic, realized according to the same principles and through the same material transformations as those typical of internal processes (such as energy transduction). The two aspects begin to be distinguishable in a particular evolutionary trend, related to the size increase of some groups of organisms whose adaptive abilities depend on motility. Here a specialized system develops, which, in the sensorimotor aspect, is decoupled from the metabolic basis, although it remains dependent on it in the self-constructive aspect. This decoupling reveals a complexification of the organization. In the last section of the article this approach to natural agency is used to analyze artificial systems by posing two problems: whether it is possible to artificially build an organization similar to the natural, and whether this notion of agency can be grounded on different organizing principles. PMID- 15811226 TI - Transient phenomena in learning and evolution: genetic assimilation and genetic redistribution. AB - Deacon has recently proposed that complexes of genes can be integrated into functional groups as a result of environmental changes that mask and unmask selection pressures. For example, many animals endogenously synthesize ascorbic acid (vitamin C), but anthropoid primates have only a nonfunctional version of the crucial gene for this pathway. It is hypothesized that the loss of functionality occurred in the evolutionary past when a diet rich in vitamin C masked the effect of the gene, and its loss effectively trapped the animals in a fruit-eating lifestyle. As a result, the complex of abilities that support this lifestyle were evolutionarily bound together, forming a multilocus complex. In this study we use evolutionary computation simulations to explore the thesis that masking and unmasking can transfer dependence from one set of genes to many sets, and thereby integrate the whole complex of genes. We used a framework based on Hinton and Nowlan's 1987 simulation of the Baldwin effect. Additional gene complexes and an environmental parameter were added to their basic model, and the fitness function extended. The simulation clearly demonstrates that the genetic redistribution effect can occur in silico, showing an initial advantage of endogenously synthesized vitamin C, followed by transfer of the fitness contribution to the complex of genes that together allow the acquisition of vitamin C from the environment. As is well known in the modeling community, the Baldwin effect only occurs in simulations when the population of agents is ''poised on the brink'' of discovering the genetically specified solution. Similarly, the redistribution effect occurs in simulations under specific initial conditions: too little vitamin C in the environment, and its synthesis it is never fully masked; too much vitamin C, and the abilities required to acquire it are not tightly integrated. The Baldwin effect has been hypothesized as a potential mechanism for developing language-specific adaptations like innate universal grammar and other highly modular capacities. We conclude with a discussion of the relevance of genetic assimilation and genetic redistribution to the evolution of language and other cognitive adaptations. PMID- 15811227 TI - Material representations: from the genetic code to the evolution of cellular automata. AB - We present a new definition of the concept of representation for cognitive science that is based on a study of the origin of structures that are used to store memory in evolving systems. This study consists of novel computer experiments in the evolution of cellular automata to perform nontrivial tasks as well as evidence from biology concerning genetic memory. Our key observation is that representations require inert structures to encode information used to construct appropriate dynamic configurations for the evolving system. We propose criteria to decide if a given structure is a representation by unpacking the idea of inert structures that can be used as memory for arbitrary dynamic configurations. Using a genetic algorithm, we evolved cellular automata rules that can perform nontrivial tasks related to the density task (or majority classification problem) commonly used in the literature. We present the particle catalogs of the new rules following the computational mechanics framework. We discuss if the evolved cellular automata particles may be seen as representations according to our criteria. We show that while they capture some of the essential characteristics of representations, they lack an essential one. Our goal is to show that artificial life can be used to shed new light on the computation-versus dynamics debate in cognitive science, and indeed function as a constructive bridge between the two camps. Our definitions of representation and cellular automata experiments are proposed as a complementary approach, with both dynamics and informational modes of explanation. PMID- 15811228 TI - Friends reunited? Evolutionary robotics and representational explanation. AB - Robotics as practiced within the artificial life community is no longer the bitter enemy of representational explanation in the way that it sometimes seemed to be in the heady, revolutionary days of the 1990s. This rapprochement is, however, fragile, because the field of evolutionary robotics continues to pose two important challenges to the idea that real-time intelligent action must or should be explained by appeal to inner representations. The first of these challenges, the threat from nontrivial causal spread, occurs when extra-neural factors account for the kind of adaptive richness and flexibility normally associated with representation-based control. The second, the threat from continuous reciprocal causation, occurs when the causal contributions made by the systemic components collectively responsible for behavior generation are massively context-sensitive and variable over time. I argue that while the threat from nontrivial causal spread can be resisted, the threat from continuous reciprocal causation provides a stern test for our representational intuitions. PMID- 15811229 TI - Beyond the flesh: some lessons from a mole cricket. AB - What do linguistic symbols do for minds like ours, and how (if at all) can basic embodied, dynamical, and situated approaches do justice to high-level human thought and reason? These two questions are best addressed together, since our answers to the first may inform the second. The key move in scaling up simple embodied cognitive science is, I argue, to take very seriously the potent role of human-built structures in transforming the spaces of human learning and reason. In particular, in this article I look at a range of cases involving what I dub surrogate situations. Here, we actively create restricted artificial environments that allow us to deploy basic perception-action-reason routines in the absence of their proper objects. Examples include the use of real-world models, diagrams, and other concrete external symbols to support dense looping interactions with a variety of stable external structures that stand in for the absent states of affairs. Language itself, I finally suggest, is the most potent and fundamental form of such surrogacy. Words are both cheap stand-ins for gross behavioral outcomes, and the concrete objects that structure new spaces for basic forms of learning and reason. A good hard look at surrogate situatedness thus turns the standard skeptical challenge on its head. But it raises important questions concerning what really matters about these new approaches, and it helps focus what I see as the major challenge for the future: how, in detail, to conceptualize the role of symbols (both internal and external) in dynamical cognitive processes. PMID- 15811230 TI - Beyond robotics: a new proactive research initiative from the EU IST program. AB - A research initiative on embodied artificial intelligence has been launched in 2004 by the Future and Emerging Technologies (FET) arm of the Information Society Technologies (IST) programme of the European Union. The initiative is called "Beyond Robotics" to emphasize that the research projects funded are required to set ambitious objectives and to aim at breakthroughs going well beyond the state of the art. Four projects worth 20 M E of EC funding were selected for 2004-2008 to address the objectives of the initiative. PMID- 15811232 TI - Cortical representations of personally familiar objects and places: functional organization of the human posterior cingulate cortex. AB - The recognition of both personally familiar objects and places involves nonspatial memory retrieval processes, but only personally familiar places are represented as space. Although the posterior cingulate cortex (PCC) is considered to process both types of such memories, its functional organization is poorly understood. In this event-related fMRI study, normal subjects judged familiar/unfamiliar pictures in four categories: familiar places (FP), familiar objects (FO), unfamiliar places (UP), and unfamiliar objects (UO), thus constituting a two-factorial design. A significant main effect of stimuli with greater activation in the place (FP and UP) than object (FO and UO) trials was observed bilaterally in several medial temporo-occipito-parietal regions, including the caudal PCC (cPCC) and parahippocampal gyrus. The reverse comparison revealed greater activation in the lateral inferior occipito-temporal junctions and intraparietal sulci bilaterally. A significant main effect of familiarity with greater activation in the familiar (FP and FO) than unfamiliar (UP and UO) trials was observed in the mid-dorsal PCC (mPCC), retrosplenial cortex, posterior precuneus, and the left intraparietal sulcus. Activation specific to the FP trials (as assessed by the interaction) was observed in the right posterodorsal PCC (pPCC) only. Together with data from previous functional imaging studies, the results suggest a functional segregation of human PCC with differential involvement of pPCC in spatial representations of personally familiar places and of the mPCC and retrosplenial cortex in episodic retrieval of personally familiar places and objects. Activation of the left intraparietal sulcus may reflect retrieval of memories related to object manipulation. PMID- 15811233 TI - Repetition-priming modulates category-related effects on event-related potentials: further evidence for multiple cortical semantic systems. AB - In the present study, the significance of category-related brain activations as an index of semantic memory structure was assessed within a repetition-priming paradigm during a lexical decision task. The interpretation of category-related effects has been debated since previous studies observed category-related brain activity mainly in tasks requiring explicit semantic categorization. Furthermore, categories were frequently associated with behavioral performance differences, which could have influenced the pattern of brain activation. Event-related potentials (ERPs) to words denoting objects from artifactual (e.g., tools) and natural categories (e.g., animals) were recorded while subjects were presented with words and pseudowords as distracters, which were repeatedly presented. Category-related ERP differences emerged in the time window of the N400, an electrophysiological index of semantic processing, over occipito-parietal and fronto-central regions as well as in the following window of a late positive potential. Repetition priming modulated these category-related ERP effects whereas behavioral repetition priming (faster reactions to repeated words) was comparable for both categories. Differences in ERP repetition effects were specifically due to diminished category-related activity at repeated presentation. The present results show that category-related brain activation is not confined to tasks requiring explicit semantic categorization. Most importantly, the study demonstrates that category-related brain activation can be specifically modulated by repetition priming in the absence of corresponding behavioral performance differences. These findings therefore substantiate the significance of category-related brain activations as reflections of semantic memory structure and support the notion of multiple cortical semantic systems. PMID- 15811234 TI - Mirror neurons responding to observation of actions made with tools in monkey ventral premotor cortex. AB - In the present study, we describe a new type of visuomotor neurons, named tool responding mirror neurons, which are found in the lateral sector of monkey ventral premotor area F5. Tool-responding mirror neurons discharge when the monkey observes actions performed by an experimenter with a tool (a stick or a pair of pliers). This response is stronger than that obtained when the monkey observes a similar action made with a biological effector (the hand or the mouth). These neurons respond also when the monkey executes actions with both the hand and the mouth. The visual and the motor responses of each neuron are congruent in that they share the same general goal, that is, taking possession of an object and modifying its state. It is hypothesized that after a relatively long visual exposure to tool actions, a visual association between the hand and the tool is created, so that the tool becomes as a kind of prolongation of the hand. We propose that tool-responding mirror neurons enable the observing monkey to extend action-understanding capacity to actions that do not strictly correspond to its motor representations. Our findings support the notion that the motor cortex plays a crucial role in understanding action goals. PMID- 15811235 TI - Pronominal reference in sentences about persons or things: an electrophysiological approach. AB - German pronouns (er(MALE/masculine), sie(FEMALE/feminine)) that refer to a person are determined by the biological gender (MALE/FEMALE) and/or syntactic gender (masculine/feminine) of the person. Pronouns (er(masculine), sie(feminine)) that refer to a thing are determined by the syntactic gender of this thing (Garten [garden]masculine, Tasche [hand-bag]feminine). The study aimed to investigate whether semantic integration, syntactic integration, or both are involved in establishing co-reference between pronoun and subject/antecedent in sentences. Here we focused on two event-related potential components: the SPS/P600, related to syntactic violation and reanalysis, and the N400 component, related to semantic integration problems. In one condition, a person was introduced as antecedent and later referred to by a pronoun, which either agreed in biological/syntactic gender or not (biological/syntactic gender violation). In a second condition, a thing was introduced as antecedent and the corresponding pronoun either agreed in syntactic gender or not (syntactic gender violation). Results at critical pronouns showed a P600 effect for incongruent compared with congruent pronouns in both conditions with a centro-parietal maximum. This effect was larger for the person compared to the thing condition. We interpreted this finding as reflecting a syntactic integration process that can be influenced by conceptual/semantic and syntactic information of the antecedent type. Furthermore, at the word following the pronoun, we observed an N400 for the thing but not for the person condition. We suggest, supported by the results of a control experiment, that this effect reflects continuous integration processes for things, whereas for persons the integration seems to be finished at pronoun position. PMID- 15811236 TI - Deferred imitation of action sequences in developmental amnesia. AB - The aims of this study were to investigate whether patients with developmental amnesia (DA) associated with bilateral hippocampal volume reduction show an impairment in incidental nonverbal recall of action sequences, and whether the severity of this memory impairment is influenced by the sequence structure (causal vs. arbitrary). Like adult-onset cases of amnesia (McDonough, Mandler, McKee, & Squire, 1995), patients with DA did not differ significantly from their age-, sex-, and IQ-matched controls in spontaneous production of the sequences prior to modeling but recalled fewer target actions and action pairs than the control group after a 24-hour delay, independent of sequence structure. Unlike the patients with adult-onset amnesia, however, the patients with DA showed some memory for both types of sequences after a 24-hour delay. This difference in severity of memory impairment might reflect differences in extent of pathology and/or age at injury. PMID- 15811237 TI - Dissociations of face and object recognition in developmental prosopagnosia. AB - Neuropsychological studies with patients suffering from prosopagnosia have provided the main evidence for the hypothesis that the recognition of faces and objects rely on distinct mechanisms. Yet doubts remain, and it has been argued that no case demonstrating an unequivocal dissociation between face and object recognition exists due in part to the lack of appropriate response time measurements (Gauthier et al., 1999). We tested seven developmental prosopagnosics to measure their accuracy and reaction times with multiple tests of face recognition and compared this with a larger battery of object recognition tests. For our systematic comparison, we used an old/new recognition memory paradigm involving memory tests for cars, tools, guns, horses, natural scenes, and houses in addition to two separate tests for faces. Developmental prosopagnosic subjects performed very poorly with the face memory tests as expected. Four of the seven prosopagnosics showed a very strong dissociation between the face and object tests. Systematic comparison of reaction time measurements for all tests indicates that the dissociations cannot be accounted for by differences in reaction times. Contrary to an account based on speed accuracy tradeoffs, prosopagnosics were systematically faster in nonface tests than in face tests. Thus, our findings demonstrate that face and nonface recognition can dissociate over a wide range of testing conditions. This is further support for the hypothesis that face and nonface recognition relies on separate mechanisms and that developmental prosopagnosia constitutes a disorder separate from developmental agnosia. PMID- 15811238 TI - A dissociation between the representation of tool-use skills and hand dominance: insights from left- and right-handed callosotomy patients. AB - The overwhelming majority of evidence indicates that the left cerebral hemisphere of right-handed humans is dominant both for manual control and the representation of acquired skills, including tool use. It is, however, unclear whether these functions involve common or dissociable mechanisms. Here we demonstrate that the disconnected left hemispheres of both right- and left-handed split-brain patients are specialized for representing acquired tool-use skills. When required to pantomime actions associated with familiar tools (Experiment 2), both patients show a right-hand (left hemisphere) advantage in response to tool names, pictures, and actual objects. Accuracy decreases as stimuli become increasingly symbolic when using the left hand (right hemisphere). Tested in isolation with lateralized pictures (Experiment 3), each patient's left hemisphere demonstrates a significant advantage over the right hemisphere for pantomiming tool-use actions with the contralateral hand. The fact that this asymmetry occurs even in a left-handed patient suggests that the left hemisphere specialization for representing praxis skills can be dissociated from mechanisms involved in hand dominance located in the right hemisphere. This effect is not attributable to differences at the conceptual level, as the left and right hemispheres are equally and highly competent at associating tools with observed pantomimes (Experiment 4). PMID- 15811239 TI - Listening to action-related sentences activates fronto-parietal motor circuits. AB - Observing actions made by others activates the cortical circuits responsible for the planning and execution of those same actions. This observation-execution matching system (mirror-neuron system) is thought to play an important role in the understanding of actions made by others. In an fMRI experiment, we tested whether this system also becomes active during the processing of action-related sentences. Participants listened to sentences describing actions performed with the mouth, the hand, or the leg. Abstract sentences of comparable syntactic structure were used as control stimuli. The results showed that listening to action-related sentences activates a left fronto-parieto-temporal network that includes the pars opercularis of the inferior frontal gyrus (Broca's area), those sectors of the premotor cortex where the actions described are motorically coded, as well as the inferior parietal lobule, the intraparietal sulcus, and the posterior middle temporal gyrus. These data provide the first direct evidence that listening to sentences that describe actions engages the visuomotor circuits which subserve action execution and observation. PMID- 15811240 TI - Transmodal sensorimotor networks during action observation in professional pianists. AB - Audiovisual perception and imitation are essential for musical learning and skill acquisition. We compared professional pianists to musically naive controls with fMRI while observing piano playing finger-hand movements and serial finger-thumb opposition movements both with and without synchronous piano sound. Pianists showed stronger activations within a fronto-parieto-temporal network while observing piano playing compared to controls and contrasted to perception of serial finger-thumb opposition movements. Observation of silent piano playing additionally recruited auditory areas in pianists. Perception of piano sounds coupled with serial finger-thumb opposition movements evoked increased activation within the sensorimotor network. This indicates specialization of multimodal auditory-sensorimotor systems within a fronto-parieto-temporal network by professional musical training. Musical ''language,'' which is acquired by observation and imitation, seems to be tightly coupled to this network in accord with an observation-execution system linking visual and auditory perception to motor performance. PMID- 15811241 TI - Sequential memory: a putative neural and synaptic dynamical mechanism. AB - A key issue in the neurophysiology of cognition is the problem of sequential learning. Sequential learning refers to the ability to encode and represent the temporal order of discrete elements occurring in a sequence. We show that the short-term memory for a sequence of items can be implemented in an autoassociation neural network. Each item is one of the attractor states of the network. The autoassociation network is implemented at the level of integrate-and fire neurons so that the contributions of different biophysical mechanisms to sequence learning can be investigated. It is shown that if it is a property of the synapses or neurons that support each attractor state that they adapt, then every time the network is made quiescent (e.g., by inhibition), then the attractor state that emerges next is the next item in the sequence. We show with numerical simulations implementations of the mechanisms using (1) a sodium inactivation-based spike-frequency-adaptation mechanism, (2) a Ca(2+)-activated K+ current, and (3) short-term synaptic depression, with sequences of up to three items. The network does not need repeated training on a particular sequence and will repeat the items in the order that they were last presented. The time between the items in a sequence is not fixed, allowing the items to be read out as required over a period of up to many seconds. The network thus uses adaptation rather than associative synaptic modification to recall the order of the items in a recently presented sequence. PMID- 15811242 TI - Brain activation during face perception: evidence of a developmental change. AB - Behavioral studies suggest that children under age 10 process faces using a piecemeal strategy based on individual distinctive facial features, whereas older children use a configural strategy based on the spatial relations among the face's features. The purpose of this study was to determine whether activation of the fusiform gyrus, which is involved in face processing in adults, is greater during face processing in older children (12-14 years) than in younger children (8-10 years). Functional MRI scans were obtained while children viewed faces and houses. A developmental change was observed: Older children, but not younger children, showed significantly more activation in bilateral fusiform gyri for faces than for houses. Activation in the fusiform gyrus correlated significantly with age and with a behavioral measure of configural face processing. Regions believed to be involved in processing basic facial features were activated in both younger and older children. Some evidence was also observed for greater activation for houses versus faces for the older children than for the younger children, suggesting that processing of these two stimulus types becomes more differentiated as children age. The current results provide biological insight into changes in visual processing of faces that occur with normal development. PMID- 15811243 TI - Preattentive binding of auditory and visual stimulus features. AB - We investigated the role of attention in feature binding in the auditory and the visual modality. One auditory and one visual experiment used the mismatch negativity (MMN and vMMN, respectively) event-related potential to index the memory representations created from stimulus sequences, which were either task relevant and, therefore, attended or task-irrelevant and ignored. In the latter case, the primary task was a continuous demanding within-modality task. The test sequences were composed of two frequently occurring stimuli, which differed from each other in two stimulus features (standard stimuli) and two infrequently occurring stimuli (deviants), which combined one feature from one standard stimulus with the other feature of the other standard stimulus. Deviant stimuli elicited MMN responses of similar parameters across the different attentional conditions. These results suggest that the memory representations involved in the MMN deviance detection response encoded the frequently occurring feature combinations whether or not the test sequences were attended. A possible alternative to the memory-based interpretation of the visual results, the elicitation of the McCollough color-contingent aftereffect, was ruled out by the results of our third experiment. The current results are compared with those supporting the attentive feature integration theory. We conclude that (1) with comparable stimulus paradigms, similar results have been obtained in the two modalities, (2) there exist preattentive processes of feature binding, however, (3) conjoining features within rich arrays of objects under time pressure and/or longterm retention of the feature-conjoined memory representations may require attentive processes. PMID- 15811244 TI - Revisiting previously searched locations in visual neglect: role of right parietal and frontal lesions in misjudging old locations as new. AB - Right-hemisphere patients with left neglect often demonstrate abnormal visual search, re-examining stimuli to the right while ignoring those to the left. But re-fixations alone do not reveal if patients misjudge whether they have searched a location before. Here, we not only tracked the eye movements of 16 neglect patients during search, but also asked them to click a response button only when they judged they were fixating a target for the very first time. ''Re-clicking'' on previously found targets would indicate that patients erroneously respond to these as new discoveries. Lesions were mapped with high-resolution MRI. Neglect patients with damage involving the right intraparietal sulcus or right inferior frontal lobe ''re-clicked'' on previously found targets on the right at a pathological rate, whereas those with medial occipito-temporal lesions did not. For the intraparietal sulcus patients, the probability of erroneous re-clicks on an old target increased with time since first discovering it; whereas for frontal patients it was independent of search time, suggesting different underlying mechanisms in these two types of patient. Re-click deficits correlated with degree of leftward neglect, mainly due to both being severe in intraparietal cases. These results demonstrate that misjudging previously searched locations for new ones can contribute to pathological search in neglect, with potentially different mechanisms being involved in intraparietal versus inferior frontal patients. When combined with a spatial bias to the right, such deficits might explain why many neglect patients often re-examine rightward locations, at the expense of items to their left. PMID- 15811245 TI - Separation of the systems for color and spatial manipulation in working memory revealed by a dual-task procedure. AB - The manipulation of different kinds of content is fundamental to working memory. It has been suggested that the mere maintenance of color and spatial information occurs in parallel, but little is known about whether this holds true for manipulation as well. Using a dual-task delayed-response paradigm that required the manipulation of color and angles, this study finds that the two functions do not interfere. Conversely, interference did occur when both components of a dual task tapped into the spatial system. Thus, color and spatial information are manipulated in parallel. A concurrent phonological task did not interfere with either maintenance or manipulation, whereas a task requiring central executive processes interfered with manipulation only. We speculate that the ventral-dorsal dissociation of visual processing is conserved for manipulation processes and that manipulation differs from maintenance in the extent to which is relies on central executive resources. PMID- 15811248 TI - Clinical management guidelines: status and issues. PMID- 15811249 TI - Pharmacogenetic study of statin therapy and cholesterol reduction. PMID- 15811250 TI - Cholesteryl ester transfer protein TaqIB polymorphism in the cholesterol and recurrent events study. PMID- 15811251 TI - Complex phenotypes and complex genetics: an introduction to genetic studies of complex traits. AB - There is currently intense interest in the genetic factors contributing to many diseases with cardiovascular complications. Diseases like atherosclerosis, diabetes, and hypertension are referred to as complex traits because multiple genes contribute to the phenotype either individually or through interactions with each other or the environment. Enabled and energized by the striking successes over the past 20 years in identifying genes that are responsible for single gene traits, many geneticists have turned to the investigation of methods that will allow for the dissection of complex traits. There have already been some successes, so there is no reason to consider the problem as inherently intractable. However, it is important to reflect on what conditions are necessary for the identification of genes that operate in complex traits. A recurring theme in this research area has been difficulty in repeating and validating research findings, and this most often can be attributed to limitations in study design. It is also important to consider that any particular research strategy can only hope to describe a portion of factors that contribute to variation in the population; therefore, the genetic approach cannot be a panacea. New efficient technologies for genotyping and public databases describing the fine structure of genetic correlations in the genome should aid many aspects of the gene discovery process. PMID- 15811252 TI - Coronary heart disease and polymorphisms in genes affecting lipid metabolism and inflammation. AB - Several biologic systems contribute to the pathophysiology of atherosclerosis and its complications, and within each of these systems many genes have been explored to establish the possible implication of their variability in coronary heart disease (CHD) risk. This report is focused on recent results pertaining to lipid and inflammatory genes, their variability, and their relationship with intermediate phenotypes and CHD. For both systems, there is no evidence at the present time that testing genetic polymorphisms might be of any benefit to the patient, for the diagnosis or prognosis of CHD, or for tailoring drug prescription. Understanding the genetics of complex traits like CHD will require a system approach that allows a modeling of the interaction among genes as well as between genetic and nongenetic sources of variation. PMID- 15811253 TI - Genetic lipoprotein disorders and coronary atherosclerosis. AB - Familial lipoprotein disorders are seen frequently in subjects with premature coronary artery disease. The genetic basis for several dyslipoproteinemias has been elucidated in the past two decades. The majority of lipoprotein disorders result from a combination of polygenic predisposition and poor lifestyle habits, including physical inactivity, increased visceral adipose tissue, increased caloric intake, and cigarette smoking. Monogenic disorders are seen in approximately 5% of premature coronary artery disease cases, and this prevalence is higher in populations with the Founder effect. Unraveling the genetic basis of many lipoprotein disorders has allowed fundamental discoveries in molecular cellular physiology and has paved the way for novel therapeutic approaches. PMID- 15811254 TI - Pharmacogenetics of response to statins: where do we stand? AB - Cardiovascular disease is one of the leading causes of death, especially in developed countries. Blood cholesterol lowering by way of statin therapy is a common risk-lowering therapy. The risk reduction for coronary artery disease for patients using statins is 27%. These reductions, however, are average effects for all patients included in the trials. There is notable interindividual variation in response to statins, and the origins of this variation are poorly understood. Pharmacogenetics seeks to determine the role of genetic factors in variation of drug response. In patients with primary hypercholesterolemia, 23 studies have examined the effects of genetic polymorphisms at 20 different loci on the lipid response to statin treatment, and 18 studies examined genetic polymorphisms involved in the benefits of statin therapy in the prevention of cardiovascular disease. Even though many studies have been performed, few results have been replicated. It is our contention that larger sample sizes and consideration of multiple genes are needed in the field of pharmacogenetics of statin response. PMID- 15811255 TI - The PEACE study: limiting the role of angiotensin-converting enzyme inhibitors in stable coronary artery disease. PMID- 15811256 TI - Nitrosylation of thiols in vascular homeostasis and disease. AB - Post-translational modification of thiols to form a nitrosothiol (S nitrosylation) is gaining attention as a mechanism by which nitric oxide can exert some of its effects in the cardiovascular system, as well as in other systems. It has been proposed that this modification would have a particularly important role in cell signaling. However, its study has been hampered by many technical difficulties. In this paper, we summarize current achievements in the field that may help to answer the questions that have been posed about the functional role of this modification. Some of these achievements have arisen from methodologic improvements, such as proteomic studies or approaches to identify subcellular localization and tissue distribution of the modification. New functional consequences of the modification of individual proteins are also summarized. PMID- 15811257 TI - The interaction of coronary tone and cardiac fibrosis. AB - Regulation of coronary vascular tone is critical for proper perfusion and function of the myocardium. Many disease processes result in compromised regulation of coronary vascular tone and impaired myocardial perfusion. A common result of coronary vascular dysfunction is the development of areas of replacement fibrosis within the myocardium and surrounding the vasculature. Both intravascular processes, such as coronary atherosclerosis and endothelial dysfunction, and extravascular processes, including compromised myocardial metabolism, hormone excesses, and altered local signaling, may result in coronary vascular dysregulation. Coronary occlusion events, in turn, lead to myocardial damage and the activation of inflammatory cells and fibroblasts. The role of fibroblasts in regulating myocardial fibrosis and the contribution of myofibroblasts, cells that have limited contractile potential while retaining many of the extracellular matrix regulating processes of the fibroblast, may also contribute to the development of myocardial disease. In this review we examine the recent literature on myocardial fibrosis and myofibroblast activity, highlighting the effects of several classes of cardiovascular agents on the remodeling process. PMID- 15811258 TI - Critical role of microenvironmental factors in angiogenesis. AB - Therapeutic angiogenesis, which entails the induction of new blood vessels by the delivery of angiogenic growth factors, is a highly attractive approach to the treatment of ischemic diseases. However, it is becoming increasingly clear that this is not easily achieved, as the effects of angiogenic growth factors can differ markedly depending on the timing of their expression, on the shape of the concentration gradients they form in vivo, and the inter-actions between endothelial cells and pericytes they induce. In fact, the same dose of vascular endothelial growth factor can induce stable, nonleaky, pericyte-covered normal capillaries or aberrant vascular structures that develop into hemangiomas. This difference in outcome can be due solely to the spatial characteristics of the delivery method. If delivery allows a homogeneous spatial distribution of VEGF in the microenvironment around each producing cell, angiogenesis can be therapeutic, whereas if the total dose is the average of diverse spatial levels, aberrant angiogenesis cannot be avoided. To achieve therapeutic angiogenesis, a means of regulating the microenvironmental levels of angiogenic factors will be critical to the generation of effective new treatment strategies. PMID- 15811259 TI - Advances in the genetic basis of coronary artery disease. AB - Exciting advances have been made recently in genetic studies of coronary artery disease (CAD), myocardial infarction (MI), and ischemic stroke. One disease causing gene for CAD and MI has been identified as MEF2A, which is located on chromosome 15q26.3 and encodes a transcriptional factor with a high level of expression in coronary endothelium. Approximately 1% to 2% of CAD patients may carry an MEF2A mutation. Four new susceptibility genes have been identified using genome-wide association studies or genome-wide linkage studies: LTA (encoding cytokine lymphotoxin-alpha) on 6p21.3 for MI; LGALS2 (encoding galectin-2, an LTA interacting protein) on 22q12-q13 for MI; ALOX5AP (encoding 5-lipoxygenase activating protein involved in synthesizing potent pro-inflammatory leukotrienes) on 13q12-13 for MI and stroke; and PDE4D (encoding phosphodiesterase 4D) on 5q12 for ischemic stroke. These studies identify a new mechanism, the myocyte enhancer factor 2 (MEF2) signaling pathway of vascular endothelium, for the pathogenesis of CAD, and also confirm the role of inflammation in the disease process. PMID- 15811261 TI - [Acute liver failure in a hospital with a liver transplantation program]. AB - INTRODUCTION: Acute hepatic failure (AHF) is an uncommon entity but with high mortality. Liver transplantation has improved prognosis but is an aggressive treatment with high risk. Currently, there are no accurate criteria to differentiate between irreversible AHF and the possibility of hepatic regeneration. The aim of this study is to review the clinical characteristics indicating transplantation and the outcome of AHF in patients at our institution. PATIENTS AND METHOD: Descriptive and retrospective study of cases of AHF at our institution in the last 3 years. RESULTS: Eleven patients were studied. Mortality was lower in transplant recipients (23%; 2 out of 8) than in non-transplanted patients (67%; 2 out 3). Hepatic regeneration occurred in one patient. Overall mortality was 36% with a perioperative mortality of 13%. CONCLUSION: The indication of hepatic transplantation in patients with a poor prognosis, early stage acute liver failure, and a low grade of encephalopathy, and prior to the development of multiorgan failure could improve the results of transplantation and reduce perioperative mortality. PMID- 15811260 TI - Oxidative stress and atherosclerosis. AB - Understanding of the pathophysiology of atherosclerosis can provide new strategies for the prevention and treatment of patients with this common disease. Clinical, epidemiologic, and basic molecular science studies have identified oxidative stress as a factor contributing to the development and progression of atherosclerosis. Oxidative stress also participates in the pathogenesis of endothelial dysfunction and hypertension, two important factors in many patients with atherosclerosis. Further, it contributes to mechanisms of disease progression such as lipid oxidation and vascular remodeling. This article reviews the role of reactive oxygen species and oxidative stress in atherosclerosis. PMID- 15811262 TI - [Gastrointestinal inflammatory fibroid polyps. Clinical characteristics and follow-up in a series of 26 patients]. AB - INTRODUCTION: Inflammatory fibroid polyp (IFP) is a protuberant lesion, located near the muscularis mucosae and composed of a proliferation of fusiform cells and conjunctive fibers surrounding capillaries and a variable inflammatory infiltrate. It is believed to be a poorly controlled inflammatory repair response. Our aim was to study the clinical, pathological and follow-up characteristics of a series of patients with IFP. PATIENTS AND METHOD: We studied 26 IFPs from 25 patients (16 women and 9 men) registered between 1985 and 2001 in a specific register of 3 centers in the city of Gerona (Spain). The variables analyzed were age, sex and clinical presentation, IFP localization and size, mucosal characteristics and associated disease, as well as follow-up information. Routine statistical analyses were performed. RESULTS: IFPs were antral in 16 patients, ileal in 7, jejunal in 2 and colonic in the remaining patient. Size determined whether they were symptomatic (35 +/- 13.6 mm) or asymptomatic (8.4 +/ 6.3 mm). Gastric polyps were significantly smaller than intestinal polyps. Symptomatic polyps (5 out of 16 gastric polyps and 9 out of 10 intestinal polyps) predominated in women and occurred at a significantly lower age than asymptomatic polyps (59.2 versus 74.1 years). Most gastric IFPs were associated with chronic atrophic gastritis while only one ileal polyp was associated with Meckels diverticulum. The mean length of follow-up was 60.6 months and, except in one patient who underwent incomplete resection, no recurrences of IFP were observed. CONCLUSION: IFP is a heterogeneous entity, depending on age at presentation, sex, size and location in the digestive tract. IFP does not recur after resection. The association of gastric IFP and chronic atrophic gastritis could suggest a modulatory effect of the mucosa on IFP growth. PMID- 15811263 TI - [Uncommon etiology of gastrointestinal bleeding: duodenal metastases from renal cell carcinoma]. AB - Because of its unpredictable behavior, renal cell carcinoma is one of the most controversial neoplasms. On the one hand, patients frequently show metastases at diagnosis because of its slight manifestations, while on the other, the neoplasm can remain stable after nephrectomy and can then metastasize many years later. When this happens, the metastases usually involve more than 2 organs. The most frequent sites of metastases are the lung and lymph nodes, followed by the bones and liver, while duodenal involvement is rare. Indeed, intestinal metastases are found in only 2% of autopsies and of these, renal cell carcinoma metastases account for 7.1%. We present a case of a solitary late recurrence presenting as upper gastrointestinal bleeding 19 years after nephrectomy for clear cell renal carcinoma. PMID- 15811264 TI - [Duodenal perforation due to an endoscopic biliary prosthesis]. AB - Endoscopic biliary drainage through endoscopic retrograde cholangiopancreatography (ERCP) is a widely accepted therapeutic option in malignant biliary obstructions. However, the procedure is not free of complications. Perforation is one possible complication although it is much less frequent (less than 1%) than pancreatitis (5.4%) or hemorrhage (2%). We present 2 cases of duodenal perforation after placement of a biliary prosthesis through ERCP. Both patients had extensive hilar cholangiocarcinoma. Onset of symptoms of perforation occurred a few hours after placement of the prosthesis and the diagnosis was confirmed by computed tomography and laparotomy. We believe that the mechanism through which perforation occurred was proximal adhesion of the prosthesis to the tumor. This increased the intensity of distal trauma produced by the intraduodenal segment, preventing adaptation of the prosthesis to intestinal peristalsis. A good preventive measure would consist of correctly adjusting the length of the prosthesis in relation to the proximal end of the biliary stenosis. PMID- 15811265 TI - [Small bowel diaphragm disease associated with NSAIDs with histology of neuromuscular and vascular hamartoma. Case report and literature review]. AB - Diaphragm disease (DD) induced by non-steroidal anti-inflammatory drugs (NSAIDs) and neuromuscular and vascular hamartoma (NMVH) are rare entities that can be difficult to recognize clinically, radiologically and pathologically. Both may cause small bowel obstruction and both reveal annular constrictions macroscopically. However, the 2 entities differ in that the constrictions are purely fibrous in DD whereas they have a hamartomatous histopathology in NMVH. We describe the case of a 76-year-old woman with a history of prolonged NSAID use who developed clinical features and gross pathology consistent with DD and histopathology of NMVH. PMID- 15811266 TI - [Acalculous cholecystitis: an uncommon form of presentation of Q fever]. AB - Acute acalculous cholecystitis is a very rare clinical presentation of Q fever. We report the case of a 38-year-old man who presented with fever associated with elevation of liver enzyme levels and thickening of the gallbladder wall on abdominal ultrasonography and who was initially diagnosed with acute acalculous cholecystitis. Due to the persistence of fever and transaminase elevation despite antibiotic treatment, a liver biopsy was performed. Characteristic "doughnut" epithelioid granulomas were observed, suggesting a diagnosis of granulomatous hepatitis caused by Q fever, which was confirmed by serological methods. Treatment with doxycycline was commenced and the patient subsequently showed rapid clinical improvement, with disappearance of fever and normalization of liver enzyme levels. We review 8 cases of acute cholecystitis associated with Q fever published in the literature and stress the importance of liver biopsy in the etiological diagnosis of patients with prolonged fever and abnormal liver function tests. PMID- 15811267 TI - [Measurement of portal pressure: a clinical parameter or research tool?]. PMID- 15811268 TI - [Joint disease in inflammatory bowel disease]. AB - The joint disorders taxonomically included in the group of seronegative spondyloarthropathies under the generic name of enteropathic arthropathy represent the most frequent extra-intestinal manifestation of inflammatory bowel disease (IBD), affecting 33% of patients. Their frequency is similar to that of ulcerative colitis and Crohn's disease. Enteropathic arthropathy consists of two main joint alterations, peripheral and axial arthritis, as well as a variable group of other peri-articular disorders. Type 1, or pauciarticular, peripheral arthritis generally coincides with IBD exacerbations, while type 2, or polyarticular, peripheral arthritis follows an independent course from IBD. Axial involvement precedes and follows an independent course from IBD and can behave as ankylosing spondylitis or asymptomatic sacroiliitis. The treatment of these rheumatologic disorders is based on the application of general measures and the use of nonsteroidal anti-inflammatory agents; intraarticular corticosteroid administration may eventually become necessary. Sulphasalazine and/or infliximab, which are indicated when the previously mentioned measures fail, can be used to treat both the articular and intestinal diseases simultaneously. PMID- 15811269 TI - [Immunological tolerance and liver transplantation]. AB - The induction of tolerance to allografts has traditionally been one of the basic aims of transplantation research. Multiple data obtained in experimental models indicate that the outcome of transplantation (rejection versus acceptance/tolerance) depends on the balance between allo-reactive cytopathic lymphocytes and immunoregulatory lymphocytes. Thus, most tolerance-inducing treatments aim to reduce the number of allo-aggressive lymphocytes and, at the same time, to increase the population of regulatory lymphocytes, which ensure graft viability once drug therapy has been withdrawn. Liver allografts are singular in that they are accepted without the need for treatment in most experimental models. Likewise, in humans, liver grafts also show a lower susceptibility to rejection than any other organ and immunosuppressive treatment can be completely eliminated in approximately 25% of recipients. Many mechanisms have been proposed to explain the tolerogenic properties of the liver. Notable among these are the effects derived from the large number of passing leukocytes present in the liver and its peculiar anatomy that maximizes contact among blood lymphocytes and liver cells with tolerogenic potential. Although there are many cases of tolerance in human allograft recipients, therapeutic strategies that would allow predictable tolerance induction and without a high risk of adverse affects are still lacking. Therefore, most studies in humans have traditionally aimed to minimize doses of immunosuppressive drugs rather than eliminate them. However, recent results in preclinical models and pilot studies indicate that therapeutic protocols for tolerance induction may become available in the not too distant future. PMID- 15811270 TI - [Metformin-induced cholestatic hepatitis]. PMID- 15811271 TI - [Colonic preparation with polyethylene glycol, hypophosphatemia and acute confusional syndrome]. PMID- 15811272 TI - [Liver transplantation in patients with HIV infection]. PMID- 15811273 TI - [Rapunzel syndrome]. PMID- 15811274 TI - [Diagnostic imaging in liver pathology]. PMID- 15811275 TI - [Macroscopic liver pathology in patients with dissociated cholestasis]. AB - OBJECTIVE: To establish the prevalence of liver focal pathology in patients with increase of alkaline phosphatase and gamma-glutamil transpeptidase and normal bilirubin (dissociated cholestasis), and to analyze the related risk factors for such pathology. METHODS: All laboratory studies of patients admitted to an Internal Medicine Department were reviewed prospectively throughout a period of 9 months. For the purpose of detecting focal liver pathology imaging liver studies (echography and/or CT) were carried out in those in which biochemical analyses showed dissociated cholestasis. RESULTS: A dissociated cholestasis pattern was found in 81 patients. In 13 of them (16%) focal liver pathology was demonstrated. The majority of the lesions (10 of 13) were local or metastatic malignant neoplasms. Sex, alcohol consumption, presence of diabetes mellitus, tumor or hepatobiliar disease previously known, or abnormalities in liver physical examination were not risk factors. No liver pathology was found in patients with an alkaline phosphatase level higher than double of gamma-glutamil transpeptidase level (sensitivity: 100%; negative predictive value: 100%). Diagnosis of a non hepatic malignant neoplasm at discharge was associated to a risk 12 times bigger for the presence of liver lesions (p < 0.01). CONCLUSIONS: It is uncommon to find focal liver pathology in patients with dissociated cholestasis. It is more common to discover focal liver pathology in patients with non-hepatic tumors and less probable when phosphatase alcaline: gamma-glutamil transpeptidase ratio is higher than two. PMID- 15811276 TI - [Carbohydrate metabolism derangements in patients with morbid obesity: weight loss impact]. AB - OBJECTIVE: Examination of carbohydrate metabolism derangements in patients with morbid obesity and the influence of weight reduction through nutritional intervention. Assessment of gender influence on carbohydrate metabolism derangements and on response to diet. PATIENTS AND METHOD: 59 patients with morbid obesity were studied with anthropometric assessment and with carbohydrate metabolism assessment through an oral glucose overload and measurement of insulin resistance, before and after a dietary intervention alternating diets of very low caloric content with conventional low-calorie diets. RESULTS: A very significant global weight reduction in both sexes was observed (95% confidence interval [CI], 26.9 to 39.8 kg in males, and 13.5 to 20.3 kg in women). A significant improvement in basal blood glucose was found in both sexes (95% CI: males, 0.44 to 46.5 mg/dl; women, 9.5 to 32.8 mg/dl), and in 120 minute oral glucose tolerance test (OGTT) blood glucose in women (95% CI: 3.5 to 52.9 mg/dl). Both males and women showed significant improvement in basal insulinemia (95% CI: males, 1.6 to 14.2 microU/ml; women, 3.9 to 8.5 microU/ml) and in sex hormone binding globulins (SHBG) (95% CI: males, -30.1 to -13.3 nmol/l; women, -54 to 19.4 nmol/l). Both males and women showed an improvement of sensitivity to insulin measured with HOMA (HOMAIR) (95% CI: males, 1.46 to 3.59; women, 0.67 to 2.24). CONCLUSION: In patients with morbid obesity carbohydrate metabolism derangements are common. Weight reduction led to a significant reduction of these derangements. PMID- 15811277 TI - [Nosocomial bacteremia: epidemiology and current antimicrobial drugs resistances]. AB - Nosocomial bacteremia is a major cause of morbidity and mortality. A change is being observed in the distribution of its causative agents, as well as an increase in the resistances to some antimicrobial drugs. We have analyzed 600 significant bacteremia episodes during the year 2002, studying the resistances to antimicrobial drugs and some epidemiological aspects. Bacteremias caused by gram positive were 57%, while the ones caused by gram-negative were 46.1%. The three microorganisms more frequently isolated were Staphylococcus epidermidis, Escherichia coli and Staphylococcus aureus. In S. aureus low resistance to oxacillin (10.3%) was observed. Streptococcus pneumoniae with lower sensitivity to penicillin was observed in 49.9%. Resistance of gram-positive microorganisms to vancomycin/teicoplanin was not observed. E. coli and Klebsiella pneumoniae showed low prevalence of expanded spectrum ss-lactamase strains. There are differences in epidemiology of bacteremias at regional level, and continuous local programs for monitoring resistances to antimicrobial drugs are needed. PMID- 15811278 TI - [Paradoxical costal shift throughout inspiration (Hoover's sign) in patients admitted because of dyspnea]. AB - OBJECTIVE: To study the frequency and diagnostic usefulness of Hoover's sign (paradoxical costal shift throughout inspiration) in patients admitted because of dyspnea. PATIENTS AND METHODS: 268 patients admitted because of dyspnea in an Internal Medicine Department were included in the study. Physical examination was carried out on the first day of admission to establish the presence of Hoover's sign. RESULTS: Hoover's sign was present in 62 patients of 82 with a diagnosis of chronic obstructive pulmonary disease (COPD) (sensitivity: 76%), in 3 patients of 101 (3%) with a diagnosis of congestive heart failure, in 3 patients of 23 (13%) with a diagnosis asthma, and in 6 patients of 62 (10%) with other diagnoses. Specificity of Hoover's sign for EPOC diagnosis was 94%. CONCLUSIONS: Hoover's sign is a frequent finding in patients admitted because of EPOC and is found only rarely in patients without obstructive pulmonary disease. This sign contributes useful information for the evaluation of patients admitted because of dyspnea. PMID- 15811279 TI - [Depressive syndromes in the immigrant population]. AB - Migrations are currently one of the most important sociocultural and political phenomena. As a rule, immigrants are in good health, although the immigration is by itself a mental health risk factor. This population shows common specific problems as adaptation processes, depressive syndromes, or other psychiatric problems. "Ulises' syndrome", depression, and dysthymia are the most common mental health conditions among unlawful non-european community immigrants cared in specific facilities. Similarly to the spanish indigenous population, anxiety disorders and readjustment disorders are the most common diagnoses among legal immigrants cared by Mental Health Services (MHS). Given the impact of sociocultural aspects in the development and clinical manifestations of mental health problems, it is necessary to know the demands of the immigrant population and to adjust current facilities for their care. PMID- 15811280 TI - [Clinical usefulness of blood pressure autoevaluation]. AB - Without a doubt blood pressure autoevaluation facilitates hypertension diagnosis, control and treatment. This also helps research, makes possible costs reduction and is an easy, safe, and useful method. Its use should be favored in the new model of patient-based clinical practice. PMID- 15811281 TI - [Therapeutic application of botulogenic toxin]. AB - Botulogenic toxin infiltration for the treatment of idiopathic local hyperhidrosis is a safe and well tolerated procedure, with minimum side effects and with an effectiveness demonstrated throughout the years, especially in axillary hyperhidrosis; in the majority of patients with this condition is an effective alternative to surgery. In other locations, the results are more modest and the clinical experience more limited, which forces us to advise the surgery as alternative with an incidence far from desirable. PMID- 15811282 TI - [Applications of monoclonal antibodies and biotechnology products in the treatment of chronic inflammatory diseases]. AB - In recent years we have noticed the arrival of biological drugs for the treatment of rheumatoid arthritis (RA), Crohn's disease (CD), psoriasis, and other chronic inflammatory diseases. Those drugs are produced with biotechnology methods and are defined as biologicals because of they work on the immune system. Different cellular groups and inflammation mediators participate in the inflammatory process, all of them susceptible of a therapeutic approach; they are so-called biological targets. Inhibition of TNF and interleukina 1 (IL-1) has proven effective for the control of inflammation in diseases as RA or CD. At present we have two types of inhibitors of TNF, specific monoclonal antibodies (infliximab, adalimumab) and cellular receptors (etanercept) and an IL-1 inhibitor (anakinra). The use of TNF inhibitors has given rise to a substantial change in the treatment of RA and CD because of its effectiveness. Together with this beneficial effect, an increase of infections (some of them severe) has occurred, especially tuberculosis. Other side effects that can be considered infrequent include demyelinization, heart failure, blood dyscrasias and lymphomas, which means that a thorough knowledge of these drugs is necessary for their use. Other potential biological drugs still in investigational phase are mentioned. PMID- 15811283 TI - [An 83-year-old women with chronic anemia, jaundice, and splenomegaly]. PMID- 15811284 TI - [Mediastinal mass in a young patient]. PMID- 15811285 TI - [Recurrent strokes and skin macules]. PMID- 15811286 TI - [Sister Mary Joseph's nodule as non-Hodgkins lymphoma clinical presentation]. PMID- 15811287 TI - [Mycobacterium marinum nodular lymphangitis]. PMID- 15811288 TI - [Citalopram in the etiology of inappropriate antidiuretic hormone secretion syndrome]. PMID- 15811289 TI - [Metallic fumes fever. With regard to a case related to manganese]. PMID- 15811291 TI - [Malaria: towards a new treatment!]. PMID- 15811292 TI - [Human Rhesus-associated glycoprotein and transmembrane transport of NH3]. PMID- 15811293 TI - [The unrestrained expansion of beta-arrestin functions spread to development]. PMID- 15811294 TI - [Fatal tuberculosis despite adaptive immune response in the absence of MyD88]. PMID- 15811295 TI - [Doping in the peloton of antituberculosis drugs]. PMID- 15811296 TI - [Lysophosphatidic acid: a new link between blood platelets and bone metastasis]. PMID- 15811297 TI - [Activating a beta-catenin signal in the liver is oncogenic]. PMID- 15811298 TI - [Pax4, islet plasticity and diabetes]. PMID- 15811300 TI - [How to localize epigenetics in the landscape of biological research?]. AB - Today, epigenetics is a very fashionable field of research. Modification of DNA by methylation, and of chromatin by histone modification or substitution represents a major fraction of the studies; but this special issue shows that epigenetic studies are very diverse, and not limited to the study of chromatin. What is common behind these different uses of the word epigenetics? A brief historical survey shows that epigenetics was invented twice, with different meanings: in the 1940s, by Conrad Waddington, as the study of the relations between the genotype and the phenotype; in the 1960s, as the global mechanisms of gene regulation involved in differentiation and development; what is common is that an approach distinct from genetics was in both cases considered as necessary because genetic models were incapable to address these problems. A good way to appreciate the relations between genetics and epigenetics is to realize that the main aim of organisms is to reproduce, and to consider the way organisms perform this task. Genetics is the precise means organisms have invented to reproduce the structure of their macromolecular components; the genome is also used to control the level and place of this reproduction. All the other means organisms have used to reproduce were more or less the result of tinkering, and constitute the field of epigenetics, with its diversity and richness. PMID- 15811301 TI - [Epigenetics as an aspect of post-genomics]. AB - A cell transmits to its progeny the activity level of many of its genes, not just their sequence. Just like the sequence may vary through a mutation, the gene activity level may change through an "epimutation" (an epigenetic modification) which is heritable and does not entail any concomitant genetic alteration. An epimutation can have important phenotypic consequences, that eventually survive to the loss of the environmental conditions that triggered it. For instance, epimutations are responsible for the divergence between a neuron and an epithelial cell that both come from the same egg and contain the same genome complement. This phenotypic difference is much larger than the one between the neurons from two animal species with dissimilar genotypes, thereby underlining the importance of epimutations. Tradition opposes the genetic and epigenetic visions, the latter being often adequated to the DNA methylation phenomenon. However, epimutations display a rich spectrum of modes that can all fit in a unique reference system based on correlated chemical, spatial and temporal scales. This reference system allows the integration of purely genetic mutations at one of its ends, thus paving the way to a new, gradual vision that encompasses the genome and the epigenome. At the other end can be found two types of epimutations that are both wide-ranging in space and rapid in producing phenotypic alterations. Firstly, long-range rearrangements of the three dimensional structure of the chromosome may influence gene expression in an heritable fashion. Such rearrangements seem to result from the collective dynamics of DNA-related activities, particularly transcription. Lastly, heritable regulatory states, e.g. a differentiated state that results from tipping a regulatory "toggle switch", involve components that are distributed throughout the nucleus or the cytoplasm, and possibly all the way to cell confines. PMID- 15811302 TI - [Epigenetics: Paramecium as a model system]. AB - Since the middle of the last century, Paramecium has appeared as an intriguing genetic model, displaying a variety of heritable characters which do not follow the Mendel laws but are cytoplasmically inherited. The analysis of the hereditary mechanisms at play in this eukaryotic unicellular organism has provided new insight into epigenetics mechanisms. Interestingly, the revealing phenomena concern two pecularities of Paramecium, its highly elaborate surface structure (with thousands of ciliary basal bodies as cytoskeleton organizers), and its nuclear dualism (coexistence of a diploid "germline" micronucleus and a highly polyploid somatic macronucleus devoted to transcription, which contains a rearranged version of the germline genome). Analysis of variant cortical organization has led to the concept of structural inheritance, implying that assembly of new organelles and supramolecular protein complexes is guided by pre existing organization. Analysis of other cytoplasmically inherited characters revealed that the developing macronucleus is epigenetically programmed by the maternal macronucleus through RNA-mediated, homology-dependent effects, suggesting the transcriptome should be recognized as a third actor in cellular inheritance, along with the "structurome" and the genome. PMID- 15811303 TI - [Variations on the topic of the "histone code"]. AB - Histones are the fundamental structural proteins intimately associated with eukaryotic DNA to form a highly ordered and condensed nucleoproteic complex termed chromatin. They are the targets of various posttranslational modifications including acetylation, methylation, phosphorylation and ubiquitination that modulate the structure/function of chromatin. The combinatorial nature of histone modifications is hypothesized to define a "histone code" that considerably extends the information potential of the genetic code, giving rise to epigenetic information. Moreover, most core histones consist of several nonallelic variants that can mark specific loci and could play an important role in establishment and maintenance of epigenetic memory. Here we will briefly present our current knowledge about histone posttranslational modifications and their implications in the regulation of epigenetic information. We will next describe core histone variants, insisting on their mode of incorporation into chromatin to discuss their epigenetic function and inheritance. PMID- 15811304 TI - [Epigenetics and development: genomic imprinting]. AB - Genomic imprinting leads to parent-of-origin-specific monoallelic expression of about 60 known genes in the mammalian genome. It was discovered 20 years ago and the aim of this review is to summarize its main characteristics. The nature of the imprint, still unknown, is characterized by differential chromatin structure and DNA methylation. The imprint is reset at each generation during gametogenesis, which can be observed by demethylation in the PGCs, then gamete specific remethylation. The imprinted genes are usually located in clusters and regulated by cis sequences such as imprinting centres, trans factors such as the insulator protein CTCF and/or large non coding antisense RNAs. Genetic and epigenetic abnormalities of the imprinted clusters can lead to human diseases such as Prader-Willi, Angelman or Beckwith-Wiedemann syndromes. PMID- 15811306 TI - [Epigenetics and cancer]. AB - Epigenetics is defined as "the study of mitotically and/or meiotically heritable changes in gene expression that cannot be explained by changes in the DNA sequence". Setting up the epigenetic program is crucial for correct development and its stable inheritance throughout its lifespan is essential for the maintenance of the tissue- and cell-specific functions of the organism. For many years, the genetic causes of cancer have hold centre stage. However, the recent wealth of information about the molecular mechanisms which, by modulating the chromatin structure, can regulate gene expression has high-lighted the predominant role of epigenetic modifications in the initiation and progression of numerous pathologies, including cancer. The nucleosome is the major target of these epigenetic regulation mechanisms. They include a series of tightly interconnected steps which starting with the setting ("writing") of the epigenetic mark till its "reading" and interpretation will result in long-term gene regulation. The major epigenetic changes associated with tumorigenesis are aberrant DNA methylation of CpG islands located in the promoter region of tumor suppressor gene, global genomic hypomethylation and covalent modifications of histone N-terminal tails which are protruding out from the nucleosome core. In sharp contrast with genetic modifications, epigenetic modifications are highly dynamic and reversible. The characterization of specific inhibitors directed against some key epigenetic players has opened a new and promising therapeutic avenue, the epigenetic therapy, since some inhibitors are already used in clinical trials. PMID- 15811305 TI - [Nutritional epigenomics of metabolic syndrome]. AB - The importance of epigenetic alterations has been acknowledged in cancer for about two decades by an increasing number of molecular oncologists who contributed to deciphering the epigenetic codes and machinery and opened the road for a new generation of drugs now in clinical trials. However, the relevance of epigenetics to common diseases such as metabolic syndrome and cardiovascular disease was less conspicuous. This review focuses on converging data supporting the hypothesis that, in addition to "thrifty genotype" inheritance, individuals with metabolic syndrome (MetS)--combining disturbances in glucose and insulin metabolism, excess of predominantly abdominally distributed weight, mild dyslipidemia and hypertension, with the subsequent development of obesity, type 2 diabetes mellitus (T2D) and cardiovascular disease (CVD)--have suffered improper "epigenetic programming" during their fetal/postnatal development due to maternal inadequate nutrition and metabolic disturbances and also during their lifetime. Moreover, as seen for obesity and T2D, MetS tends to appear earlier in childhood, to be more severe from generation to generation and to affect more pregnant women. Thus, in addition to maternal effects, MetS patients may display "transgenerational effects" via the incomplete erasure of epigenetic marks endured by their parents and grandparents. We highlight the susceptibility of epigenetic mechanisms controlling gene expression to environmental influences due to their inherent malleability, emphasizing the participation of transposable elements and the potential role of imprinted genes during critical time windows in epigenetic programming, from the very beginning of development throughout life. Increasing our understanding on epigenetic patterns significance and small molecules (nutrients, drugs) that reverse epigenetic (in)activation should provide us with the means to "unlock" silenced (enhanced) genes, and to "convert" the obsolete human thrifty genotype into a "squandering" phenotype. PMID- 15811307 TI - [Reprogramming and epigenesis]. AB - The fact that the nucleus of a differentiated somatic cell can be reprogrammed in order to sustain embryonic development is now well established. Experiments of somatic cell nuclear transfer (cloning) have proved that a foreign nucleus introduced into an enucleated oocyte can give rise to physiologically normal offsprings, with a normal lifespan. Such evidence of genome expression plasticity is also observed experimentally with heterokaryons, created by the fusion or the nuclear transfer between two somatic cells, where differentiated nuclei are able to express genes characteristic of the host cell. However, the epigenetic mechanisms that permit nuclear plasticity remain poorly understood. In this paper we present the main evidences showing important modifications of the large scale organisation of chromosomal domains and of the DNA methylation pattern upon nuclear transfer and during the first cleavages. These modifications of epigenetic marks, brought by an intimate contact between the chromatin and the recipient oocyte cytoplasmic factors, appear essential for further development. They are established over the first cell cycles of development. The onset of embryonic genome activation and the first cellular differentiation events that occur over the implantation period are two additional check-points of reprogramming that appear to be also highly dependent on epigenetic alterations. Beyond those stages, defective placental functions might be directly responsible for the fetal and postnatal physiopathologies frequently observed in cloned animals. No direct link between preimplantation reprogramming defaults, placental dysfunctions and low development to term has been established yet. The epigenetics studies which are now used to characterise loci specific and probably genotype dependent alterations in cloned animals of different species will provide invaluable help to define the role of epigenesis in the achievement of a developmental program. PMID- 15811308 TI - [Epigenetic control, development and natural genetic variation in plants]. AB - Plant life strategies differ radically from those of most animals. Plants are not motile, and can only face stress by developing appropriate physiological responses. In addition, many developmental decisions take place during post embryonic life in plants, whereas vertebrate and invertebrate development is nearly complete by the time of birth. For instance, while the germ line is typically set aside early during embryogenesis in animals, plants produce gametes from stem cell populations that were previously used for the vegetative growth of shoots. Nevertheless, plants and animals have similar nuclear organization, chromatin constitution and gene content, which raises the question as to whether or not fundamental differences in the use of genetic information underlie their distinct life strategies. More specifically, we would like to know if chromatin and the epigenetically defined, heritable cell fates that it can confer play comparable roles in plants and animals. Here we review our current knowledge on chromatin-mediated epigenetic processes in plants. Based on available evidence, we argue that epigenetic regulation of gene expression plays a relatively minor role in plants compared to mammals. Conversely, plants appear to be more prone than other multicellular organisms to the induction of chromatin-based, epigenetically modified gene activity states that can be transmitted over many generations. These so-called "epimutations" may therefore represent a significant proportion of the natural genetic variation seen in plants. In humans, epimutations are frequently observed in cancers, and given their metastable nature, they could also play an important role in familial disorders that do not demonstrate clear Mendelian inheritance. PMID- 15811309 TI - [Brain synaptogenesis and epigenesis]. AB - Synaptic plasticity, or epigenesis, is present and varies throughout the whole life of the cerebral cortex. The adult synapse is formed of large and variable proteins assemblies acting as molecular switches leading to many distinct functional states. In the flow of activity circulating through the synaptic circuits, these multiple synaptic states transitions are modulated by the levels and sequences of activations of the pre- and post-synaptic domains. The efficiency of synaptic transmission is also modulated by competition and/or cooperativity with neighbouring synapses, and by many neuromodulations. Some transitions eventually lead to synaptogenesis. In the adult cerebral cortex, synaptogenesis remains a local event; axonal and dendritic arbors are not reshaped. On the contrary, during pre- and post-natal synaptogenesis, the same molecular mechanisms lead to a significant reorganization of the axonal and dendritic arbors. Early in the development, synapses are generated and differentiate under the control of robust mechanisms governed by genes. Then, during the critical periods, extending from the end of gestation to the end of puberty, the refinement of the synaptic architecture becomes experience expectant. This "epigenetic opening" of synaptogenesis to environment is maximal in the human brain. It is the source of the exceptional cognitive adaptability of our species, and possibly one of its major fragility. Epigenetic manipulations of these critical periods are undertaken, allowing restoration of synaptic plasticity also in the adult brain. PMID- 15811310 TI - [Dopamine D3 receptors as a novel target for improving the treatment of schizophrenia]. AB - Schizophrenia is a complex and serious disorder which affects some 0.5-1.0% of the population. The disease generally begins in adolescence. This early onset, together with the progressive and often irreversible nature of schizophrenia, account for its high social cost. Positive symptoms, such as hallucinations, are generally well-controlled by antipsychotics, whereas cognitive and deficit symptoms are poorly-treated. All antipsychotic agents, irrespective of their overall receptor-binding profiles, interact with dopaminergic mechanisms that are known to be perturbed in schizophrenic patients. Dopamine exerts its actions via five classes of receptor, offering a broad palette of targets for the conception of novel antipsychotic agents. The present article focuses on the relevance of dopamine D3 receptors to the aetiology and treatment of schizophrenia. Experimental studies suggest that, as compared to other drugs, antipsychotic agents which preferentially block D3 receptors may possess therapeutic advantages, notably in the control of cognitive symptoms. The first clinical studies for the evaluation of this hypothesis have recently got underway. PMID- 15811311 TI - [Small treatise on art of warfare... from Shigella]. PMID- 15811312 TI - Hailey-Hailey disease as an orthodisease of PMR1 deficiency in Saccharomyces cerevisiae. AB - The term orthodisease has recently been introduced to define human disorders in which the pathogenic gene has orthologs in model organism genomes. Here, we describe Hailey-Hailey disease (HHD), a blistering skin disorder caused by haploinsufficiency of ATP2C1 as an orthodisease from a Saccharomyces cerevisiae perspective. ATP2C1 encodes the human secretory pathway Ca(2+)/Mn(2+) ATPase hSPCA1 and is orthologous to the PMR1 gene in S. cerevisiae. hSPCA1 fully complements PMR1 deficiency in yeast and pmr1DeltaS. cerevisiae has proved to be a valuable tool to screen ATP2C1 mutations and address potential pathogenic/pharmacologic mechanisms in HHD. Consequently, this human skin disorder is an ideal example of an orthodisease. PMID- 15811313 TI - Simulating redox coupled proton transfer in cytochrome c oxidase: looking for the proton bottleneck. AB - Gaining a detailed understanding of the molecular nature of the redox coupled proton transfer in cytochrome c oxidase (COX) is one of the challenges of modern biophysics. The present work addresses this by integrating approaches for simulations of proton transport (PTR) and electron transfer (ET). The resulting method converts the electrostatic energies of different charge configurations and reorganization energies to free-energy profiles for different PTR and ET pathways. This approach provides for the first time a tool to study the actual activation barriers and kinetics of different feasible PTR processes in the cycle of COX. Using this tool, we explore the PTR through the bottleneck water molecules. It is found that a stepwise PTR along this commonly assumed path leads to far too high barriers and is, thus, inconsistent with the observed kinetics. Furthermore, the simulated free-energy profile does not provide a simple gating mechanism. Fortunately, we obtain reasonable kinetics when we consider a PTR that involves a concerted transfer of protons to and from E286. Finally, semi qualitative considerations of the forward and backward barriers point toward open questions about the actual gating process and offer a feasible pumping mechanism. Although further studies are clearly needed, we believe that our approach offers a general and effective tool for correlating the structure of COX with its function. PMID- 15811314 TI - Inflamm-ageing and lifelong antigenic load as major determinants of ageing rate and longevity. AB - Immunosenescence is the consequence of the continuous attrition caused by chronic antigenic stress. The most important characteristics of immunosenescence (accumulation of memory and effector T cells, reduction of naive T cells, shrinkage of T cell repertoire, reduction of the immunological space) are compatible with this assumption. Immunosenescence can be taken as proof that the beneficial effects of the immune system, devoted to the neutralization of harmful agents early in life, become detrimental late in life, in a period not foreseen by evolution. This perspective could explain the mechanisms of the ageing process as well as the pathogenesis of age-related diseases. PMID- 15811315 TI - Thermolabile phenotype of carnitine palmitoyltransferase II variations as a predisposing factor for influenza-associated encephalopathy. AB - To assess the etiology of influenza-associated encephalopathy (IAE), a surveillance effort was conducted during 2000-2003 in South-West Japan. All fatal and handicapped patients except one (4/34 patients) exhibited a disorder of mitochondrial beta-oxidation evoked by the inactivated carnitine palmitoyltransferase II (CPT II) with transiently elevated serum acylcarnitine ratios (C(16:0) + C(18:1))/C(2) > 0.09 during high-grade fever. Analyses of genotypes and allele compositions of CPT II revealed a thermolabile phenotype of compound heterozygotes for [1055T > G/F352C] and [1102G > A/V368I], which shows a higher frequency in IAE patients than healthy volunteers (P < 0.025). The thermolabile phenotype of CPT II variations may be a principal genetic background of IAE in Japanese. PMID- 15811316 TI - Functional role of AMP-activated protein kinase in the heart during exercise. AB - AMP-activated protein kinase (AMPK) plays a critical role in maintaining energy homeostasis and cardiac function during ischemia in the heart. However, the functional role of AMPK in the heart during exercise is unknown. We examined whether acute exercise increases AMPK activity in mouse hearts and determined the significance of these increases by studying transgenic (TG) mice expressing a cardiac-specific dominant-negative (inactivating) AMPKalpha2 subunit. Exercise increased cardiac AMPKalpha2 activity in the wild type mice but not in TG. We found that inactivation of AMPK did not result in abnormal ATP and glycogen consumption during exercise, cardiac function assessed by heart rhythm telemetry and stress echocardiography, or in maximal exercise capacity. PMID- 15811317 TI - Xestospongin B, a competitive inhibitor of IP3-mediated Ca2+ signalling in cultured rat myotubes, isolated myonuclei, and neuroblastoma (NG108-15) cells. AB - Xestospongin B, a macrocyclic bis-1-oxaquinolizidine alkaloid extracted from the marine sponge Xestospongia exigua, was highly purified and tested for its ability to block inositol 1,4,5-trisphosphate (IP(3))-induced Ca(2+) release. In a concentration-dependent manner xestospongin B displaced [(3)H]IP(3) from both rat cerebellar membranes and rat skeletal myotube homogenates with an EC(50) of 44.6 +/- 1.1 microM and 27.4 +/- 1.1 microM, respectively. Xestospongin B, depending on the dose, suppressed bradykinin-induced Ca(2+) signals in neuroblastoma (NG108 15) cells, and also selectively blocked the slow intracellular Ca(2+) signal induced by membrane depolarization with high external K(+) (47 mM) in rat skeletal myotubes. This slow Ca(2+) signal is unrelated to muscle contraction, and involves IP(3) receptors. In highly purified isolated nuclei from rat skeletal myotubes, Xestospongin B reduced, or suppressed IP(3)-induced Ca(2+) oscillations with an EC(50) = 18.9 +/- 1.35 microM. In rat myotubes exposed to a Ca(2+)-free medium, Xestospongin B neither depleted sarcoplasmic reticulum Ca(2+) stores, nor modified thapsigargin action and did not affect capacitative Ca(2+) entry after thapsigargin-induced depletion of Ca(2+) stores. Ca(2+)-ATPase activity measured in skeletal myotube homogenates remained unaffected by Xestospongin B. It is concluded that xestospongin B is an effective cell permeant, competitive inhibitor of IP(3) receptors in cultured rat myotubes, isolated myonuclei, and neuroblastoma (NG108-15) cells. PMID- 15811318 TI - Development of highly stable galectins: truncation of the linker peptide confers protease-resistance on tandem-repeat type galectins. AB - Galectin-9 and galectin-8, members of beta-galactoside-binding animal lectin family, are promising agents for the treatment of immune-related and neoplastic diseases. The proteins consist of two carbohydrate recognition domains joined by a linker peptide, which is highly susceptible to proteolysis. To increase protease resistance, we prepared mutant proteins by serial truncation of the linker peptide. As a result, mutant forms lacking the entire linker peptide were found to be highly stable against proteolysis and retained their biological activities. These mutant proteins might be useful tools for analyzing the biological functions and evaluating the therapeutic potential of galectin-9 and galectin-8. PMID- 15811319 TI - Targeting protein homodimerization: a novel drug discovery system. AB - To identify a novel class of antibiotics, we have developed a high-throughput genetic system for targeting the homodimerization (HD system) of histidine kinase (HK), which is essential for a bacterial signal transduction system (two component system, TCS). By using the HD system, we screened a chemical library and identified a compound, I-8-15 (1-dodecyl-2-isopropylimidazole), that specifically inhibited the dimerization of HK encoded by the YycG gene of Staphylococcus aureus and induced concomitant bacterial cell death. I-8-15 also showed antibacterial activity against MRSA (methicillin-resistant S. aureus) and VRE (vancomycin-resistant Enterococcus faecalis) with MICs at 25 and 50 microg/ml, respectively. PMID- 15811320 TI - The low energy emitting states of the Lhca4 subunit of higher plant photosystem I. AB - The selectively red excited emission spectrum, at room temperature, of the in vitro reconstituted Lhca4, has a pronounced non-equilibrium distribution, leading to enhanced emission from the directly excited low-energy pigments. Two different emitting forms (or states), with maximal emission at 713 and 735nm (F713 and F735) and unusual spectral properties, have been identified. Both high-energy states are populated when selective excitation is into the F735 state and the fluorescence anisotropy spectrum attains the value of 0.3 in the wavelength region where both emission states are present. This indicates that the two states are on the same Lhca4 complex and have transition dipoles with similar orientation. PMID- 15811321 TI - Control of the Ralstonia solanacearum Type III secretion system (Hrp) genes by the global virulence regulator PhcA. AB - Expression of several virulence factors in the plant pathogen bacterium Ralstonia solanacearum is controlled by a complex regulatory network, at the center of which is PhcA. We provide genetic evidence that PhcA also represses the expression of hrp genes that code for the Type III protein secretion system, a major pathogenicity determinant in this bacterium. The repression of hrp genes in complete medium is relieved in a phcA mutant and two distinct signals, a quorum sensing signal and complex nitrogen sources, appear to trigger this PhcA dependent repression. This control of hrp gene expression by PhcA is realized at the level of the HrpG regulatory protein. PMID- 15811322 TI - The esterase from Alicyclobacillus acidocaldarius as a reporter enzyme and affinity tag for protein biosynthesis. AB - Esterase from thermophilic bacteria Alicyclobacillus acidocaldarius can be produced up to 200 microg/ml by coupled in vitro transcription/translation system derived from Escherichia coli. The synthesized thermostable enzyme can be determined by photometrical and fluorescent assays at least up to 10(-8) M concentration or by activity staining in the polyacrylamide gels. Enhanced green fluorescence protein-esterase fusion protein was bound to a matrix with immobilized esterase inhibitor and purified by affinity chromatography. Thus, the esterase is suited as a reporter enzyme to monitor the expression of polypeptides coupled to its N-terminus and simultaneously, as a cleavable tag for polypeptide purification. PMID- 15811323 TI - A novel hypertonicity-induced cation channel in primary cultures of human hepatocytes. AB - In whole-cell recordings on primary cultures of human hepatocytes, we observe the hypertonic activation of a novel type of cation channel with a permeability ratio for Na(+):Li(+):K(+):Cs(+):NMDG(+) of 1:1.2:1.3:1.2:0.6. With a P(Ca)/P(Na) of 0.7 the channel is also clearly permeable to Ca(++). Most likely, the channel is Cl(-) impermeable but its activity critically depends on the extracellular Cl(-) concentration (with the half maximal effect at 88 mmol/l). With a 64% inhibition by amiloride and a complete block by flufenamate and Gd(3+) (at 100 micromol/l each), the channel may represent a molecular link between the amiloride-sensitive and insensitive channels reported so far. PMID- 15811324 TI - N-terminal glycine-specific protein conjugation catalyzed by microbial transglutaminase. AB - Here, we report the N-terminal glycine (Gly) residue of a target protein can be a candidate primary amine for site-specific protein conjugation catalyzed by microbial transglutaminase (MTG) from Streptomyces mobaraensis. Gly5-enhanced green fluorescent protein (EGFP) (EGFP with five additional Gly residues at its N terminus) was cross-linked with Myc-dihydrofolate reductase (DHFR) (DHFR with the myc epitope sequence at its N-terminus) to yield DHFR-EGFP heterodimers. The reactivities of additional peptidyl linkers were investigated and the results obtained suggested that at least three additional Gly residues at the N-terminus were required to yield the EGFP-DHFR heterodimeric form. Site-directed mutagenesis analysis revealed marked preference of MTG for amino acids adjacent to the N-terminal Gly residue involved in the protein conjugation. In addition, peptide-protein conjugation was demonstrated by MTG-catalyzed N-terminal Gly specific modification of a target protein with the myc epitope peptide. PMID- 15811325 TI - BLG-e1 - a novel regulatory element in the distal region of the beta lactoglobulin gene promoter. AB - beta-Lactoglobulin (BLG) is a major ruminant milk protein. A regulatory element, termed BLG-e1, was defined in the distal region of the ovine BLG gene promoter. This 299-bp element lacks the established cis-regulatory sequences that affect milk-protein gene expression. Nevertheless, it alters the binding of downstream BLG sequences to histone H4 and the sensitivity of the histone-DNA complexes to trichostatin A treatment. In mammary cells cultured under favorable lactogenic conditions, BLG-e1 acts as a potent, position-independent silencer of BLG/luciferase expression, and similarly affects the promoter activity of the mouse whey acidic protein gene. Intragenic sequences upstream of BLG exon 2 reverse the silencing effect of BLG-e1 in vitro and in transgenic mice. PMID- 15811326 TI - Direct interaction and cooperative role of tumor suppressor p16 with band 3 (AE1). AB - By using the C-terminal 112-residue of band 3 to screen the K562 cDNA library, we find that the p16 interacts with band 3, which was confirmed both in yeast and in mammalian cells. Functional experiments show that p16 facilitates the movement of band 3 to plasma membrane with increased anion transport activity in 293t cells. Moreover, expression of endogenous p16 in 293t cells was increased at 24 and 36 h after transfection with band 3. Our findings provide a novel regulation pathway for both band 3 and p16. PMID- 15811327 TI - Acetylcholine from vagal stimulation protects cardiomyocytes against ischemia and hypoxia involving additive non-hypoxic induction of HIF-1alpha. AB - Electrical stimulation of the vagal efferent nerve improves the survival of myocardial infarcted rats. However, the mechanism for this beneficial effect is unclear. We investigated the effect of acetylcholine (ACh) on hypoxia-inducible factor (HIF)-1alpha using rat cardiomyocytes under normoxia and hypoxia. ACh posttranslationally regulated HIF-1alpha and increased its protein level under normoxia. ACh increased Akt phosphorylation, and wortmannin or atropine blocked this effect. Hypoxia-induced caspase-3 activation and mitochondrial membrane potential collapse were prevented by ACh. Dominant-negative HIF-1alpha inhibited the cell protective effect of ACh. In acute myocardial ischemia, vagal nerve stimulation increased HIF-1alpha expression and reduced the infarct size. These results suggest that ACh and vagal stimulation protect cardiomyocytes through the PI3K/Akt/HIF-1alpha pathway. PMID- 15811328 TI - Expression of tau reduces secretion of Abeta without altering the amyloid precursor protein content in CHOsw cells. AB - Insoluble deposits of tau and amyloid precursor protein (APP) peptides Abeta characterize Alzheimer's disease. We studied the role of tau in the metabolism of APP in cells stably expressing APP Swedish mutation (CHOsw). Transient expression of tau in CHOsw cells caused morphological changes, bundling of microtubules and perinuclear aggregation of Golgi-derived vesicles. It also reduced the secretion of Abeta(1-40) and Abeta(1-42) without altering the APP steady state levels. This was accompanied by a reduction in the gamma-secretase and an increase in the insulin degrading enzyme activities. Our results suggest that tau may play an inhibitory role in the amyloidogenic activity of APP. PMID- 15811329 TI - The cyanobacterium Gloeobacter violaceus PCC 7421 uses bacterial-type phytoene desaturase in carotenoid biosynthesis. AB - Carotenoid composition and its biosynthetic pathway in the cyanobacterium Gloeobacter violaceus PCC 7421 were investigated. beta-Carotene and (2S,2'S) oscillol 2,2'-di(alpha-L-fucoside), and echinenone were major and minor carotenoids, respectively. We identified two unique genes for carotenoid biosynthesis using in vivo functional complementation experiments. In Gloeobacter, a bacterial-type phytoene desaturase (CrtI), rather than plant-type desaturases (CrtP and CrtQ), produced lycopene. This is the first demonstration of an oxygenic photosynthetic organism utilizing bacterial-type phytoene desaturase. We also revealed that echinenone synthesis is catalyzed by CrtW rather than CrtO. These findings indicated that Gloeobacter retains ancestral properties of carotenoid biosynthesis. PMID- 15811330 TI - Synthetic peptides derived from SARS coronavirus S protein with diagnostic and therapeutic potential. AB - The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is an important viral structural protein. Based on bioinformatics analysis, 10 antigenic peptides derived from the S protein sequence were selected and synthesized. The antigenicity and immunoreactivity of all the peptides were tested in vivo and in vitro. Four peptides (P6, P8, P9 and P10) which contain B cell epitopes of the S protein were identified, and P8 peptide was confirmed in vivo to have a potential in serological diagnosis. By using a syncytia formation model, we tested the neutralization ability of all 10 peptides and their corresponding antibodies. It is interesting to find that P8 and P9 peptides inhibited syncytia formation, suggesting that the P8 and P9 spanning regions may provide a good target for anti-SARS-CoV drug design. Our data suggest that we have identified peptides derived from the S protein of SARS-CoV, which are useful for SARS treatment and diagnosis. PMID- 15811331 TI - Psychophysiological stress-regulated gene expression in mice. AB - Eight genes showed significant changes in expression in mice under psychophysiological stress provided by cage-restraint and water-immersion. The transcription level of most of these genes was affected in all the tissues analyzed, and some of them were responsive genes in several different stress systems. Peculiarly, the expression level of one gene, cdc2-like kinase 1 (CLK1), was reduced only in the brain, while the balance of partially- and alternatively spliced CLK1 mRNA species changed in all the tissues including the brain. These results suggest that some stress-response mechanisms, including transcriptional and post-transcriptional events, are coordinated in the whole body in mice under psychophysiological stress. PMID- 15811332 TI - Evaluation of nuclear transfer and transcription of plasmid DNA condensed with protamine by microinjection: the use of a nuclear transfer score. AB - In the present study, the nuclear delivery of a green fluorescence protein (GFP) encoding pDNA condensed by protamine was investigated in terms of trans-gene expression after cytoplasmic (E(cyt)) and nuclear (E(nuc)) microinjection. To compare the nuclear transfer process, a novel parameter; the nuclear transfer (NT) score was introduced. The E(cyt) value for protamine/pDNA particles increased in a charge ratio-dependent manner. The calculated NT score showed that this increase results from an enhancement in nuclear transfer efficiency, which was also quantitatively confirmed by a recently developed confocal image-assisted three-dimensionally integrated quantification (CIDIQ) method. Moreover, E(nuc) for protamine/pDNA particles was significantly higher than that for poly-L lysine/pDNA particles, suggesting that pDNA, when condensed with protamine, is more accessible to intra-nuclear transcription. Collectively, protamine is an excellent DNA condenser, with bi-functional advantages: improvement in nuclear delivery and efficient intra-nuclear transcription. PMID- 15811333 TI - Sensitization of stefin B-deficient thymocytes towards staurosporin-induced apoptosis is independent of cysteine cathepsins. AB - Stefin B (cystatin B) is an inhibitor of lysosomal cysteine cathepsins and does not inhibit cathepsin D, E (aspartic) or cathepsin G (serine) proteinases. In this study, we have investigated apoptosis triggered by camptothecin, staurosporin (STS), and anti-CD95 monoclonal antibody in the thymocytes from the stefin B-deficient mice and wild-type mice. We have observed increased sensibility to STS-induced apoptosis in the thymocytes of stefin B-deficient mice. Pretreatment of cells with pan-caspase inhibitor z-Val-Ala-Asp(OMe) fluoromethylketone completely inhibited phosphatidylserine externalization and caspase activation, while treatment with inhibitor of calpains- and papain-like cathepsins (2S,3S)-trans-epoxysuccinyl-leucylamido-3-methyl-butane ethyl ester did not prevent caspase activation nor phosphatidylserine exposure. We conclude that sensitization to apoptosis induced by STS in thymocytes of stefin B deficient and wild-type mice is not dependent on cathepsin inhibition by stefin B. PMID- 15811334 TI - Efficient suppression of the amber codon in E. coli in vitro translation system. AB - An mRNA encoding the esterase from Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA(Ser(CUA)) was monitored by determination of the full-length and active esterase. It was shown that commonly used increase of suppressor tRNA concentration inhibits protein production and therefore limits suppression. In situ deactivation of release factor by specific antibodies leads to efficient suppression already at low suppressor tRNA concentration and allows an in vitro synthesis of fully active enzyme in high yield undistinguishable from wild-type protein. PMID- 15811335 TI - Cardiolipin synthase of Arabidopsis thaliana. AB - Functional expression studies in microorganisms showed that the Arabidopsis thaliana gene At4g04870 represents the cardiolipin synthase (CLS) gene encoding a hydrophobic preprotein of 38 kDa with a cleavable signal peptide for the import into mitochondria. CLS of Arabidopsis over-expressed in Escherichia coli has an alkaline pH optimum, a strict requirement for divalent cations and a distinctly lower K(m) for cytidinediphosphate-diacylglycerol than for phosphatidylglycerol. It displayed a preference for both its substrates esterified with unsaturated acyl groups. Solubilization and purification experiments revealed that the protein requires a defined phospholipid environment, particularly the presence of cardiolipin, to acquire its catalytically active conformation. PMID- 15811336 TI - Nav1.6 channels generate resurgent sodium currents in spinal sensory neurons. AB - The Na(v)1.6 voltage-gated sodium channel has been implicated in the generation of resurgent currents in cerebellar Purkinje neurons. Our data show that resurgent sodium currents are produced by some large diameter dorsal root ganglion (DRG) neurons from wild-type mice, but not from Na(v)1.6-null mice; small DRG neurons do not produce resurgent currents. Many, but not all, DRG neurons transfected with Na(v)1.6 produce resurgent currents. These results demonstrate for the first time the intrinsic ability of Na(v)1.6 to produce a resurgent current, and also show that cell background is critical in permitting the generation of these currents. PMID- 15811337 TI - Drosophila molting neurohormone bursicon is a heterodimer and the natural agonist of the orphan receptor DLGR2. AB - Bursicon is a neurohumoral agent responsible for tanning and hardening of the cuticle and expansion of the wings during the final phase of insect metamorphosis. Although the hormonal activity was described more than 40 years ago, the molecular nature of bursicon has remained elusive. We identify here Drosophila bioactive bursicon as a heterodimer made of two cystine knot polypeptides. This conclusion was reached in part from the unexpected observation that in the genome of the honey bee, the orthologs of the two Drosophila proteins are predicted to be fused in a single open reading frame. The heterodimeric Drosophila protein displays bursicon bioactivity in freshly enclosed neck-ligated flies and is the natural agonist of the orphan G protein-coupled receptor DLGR2. PMID- 15811338 TI - Aftiphilin is a component of the clathrin machinery in neurons. AB - Aftiphilin was identified through a database search for proteins containing binding motifs for the gamma-ear domain of clathrin adaptor protein 1 (AP-1). Here, we demonstrate that aftiphilin is expressed predominantly in brain where it is enriched on clathrin-coated vesicles. In addition to eight gamma-ear-binding motifs, aftiphilin contains two WXXF-acidic motifs that mediate binding to the alpha-ear of clathrin adaptor protein 2 (AP-2) and three FXXFXXF/L motifs that mediate binding to the alpha- and beta2-ear. We demonstrate that aftiphilin uses these motifs for interactions with AP-1 and AP-2 and that it immunoprecipitates these APs but not AP-3 or AP-4 from brain extracts. Aftiphilin demonstrates a brefeldin A sensitive localization to the trans-Golgi network in hippocampal neurons where it co-localizes with AP-1. Aftiphilin is also found at synapses where it co-localizes with synaptophysin and AP-2. Our data suggest a role for aftiphilin in clathrin-mediated trafficking in neurons. PMID- 15811339 TI - Assembly of hereditary amyloid beta-protein variants in the presence of favorable gangliosides. AB - The mechanisms underlying regional amyloid beta-protein (Abeta) deposition in brain remain unclear. Here we show that assembly of hereditary variant Dutch- and Italian-type Abetas, and Flemish-type Abeta was accelerated by GM3 ganglioside, and GD3 ganglioside, respectively. Notably, cerebrovascular smooth muscle cells, which compose the cerebral vessel wall at which the Dutch- and Italian-type Abetas deposit, exclusively express GM3 whereas GD3 is upregulated in the co culture of endothelial cells and astrocytes, which forms the cerebrovascular basement membrane, the site of Flemish-type Abeta deposition. Our results suggest that regional Abeta deposition is induced by the local gangliosides in the brain. PMID- 15811340 TI - Identification of the reactive cysteine residues in oligopeptidase B from Trypanosoma brucei. AB - Oligopeptidase B (OpdB) from Trypanosoma brucei is a candidate therapeutic target in African trypanosomiasis. OpdB is an atypical serine peptidase, since activity is inhibited by thiol-blocking reagents and enhanced by reducing agents. We have identified C256 as the reactive cysteine residue that mediates OpdB inhibition by N-ethylmaleimide and iodoacetic acid. Modeling studies suggest that C256 adducts occlude the P(1) substrate-binding site, preventing substrate binding. We further demonstrate that C559 and C597 are responsible for the thiol-enhancement of OpdB activity. These studies may facilitate the development of specific OpdB inhibitors with therapeutic potential, by exploiting these unique properties of this enzyme. PMID- 15811341 TI - Confinement and crowding effects on tryptophan synthase alpha2beta2 complex. AB - Biological molecules experience in vivo a highly crowded environment. The investigation of the functional properties of the tryptophan synthase alpha(2)beta(2) complex either entrapped in wet nanoporous silica gels or in the presence of the crowding agents dextran 70 and ficoll 70 indicates that the rates of the conformational transitions associated to catalysis and regulation are reduced, and an open and less catalytically active conformation is stabilized. PMID- 15811342 TI - Histone demethylation catalysed by LSD1 is a flavin-dependent oxidative process. AB - Lysine-specific histone demethylase 1 (LSD1) is a very recently discovered enzyme which specifically removes methyl groups from Lys4 of histone 3. We have addressed the functional properties of the protein demonstrating that histone demethylation involves the flavin-catalysed oxidation of the methylated lysine. The nature of the substrate that acts as the electron acceptor required to complete the catalytic cycle was investigated. LSD1 converts oxygen to hydrogen peroxide although this reactivity is not as pronounced as that of other flavin dependent oxidases. Our findings raise the possibility that in vivo LSD1 might not necessarily function as an oxidase, but it might use alternative electron acceptors. PMID- 15811343 TI - Biochemical characterization of Neurospora crassa glycogenin (GNN), the self glucosylating initiator of glycogen synthesis. AB - Glycogenin acts in the initiation step of glycogen biosynthesis by catalyzing a self-glucosylation reaction. In a previous work [de Paula et al., Arch. Biochem. Biophys. 435 (2005) 112-124], we described the isolation of the cDNA gnn, which encodes the protein glycogenin (GNN) in Neurospora crassa. This work presents a set of biochemical and functional studies confirming the GNN role in glycogen biosynthesis. Kinetic experiments showed a very low GNN K(m) (4.41 microM) for the substrate UDP-glucose. Recombinant GNN was produced in Escherichia coli and analysis by mass spectroscopy identified a peptide containing an oligosaccharide chain attached to Tyr196 residue. Site-directed mutagenesis and functional complementation of a Saccharomyces cerevisiae mutant strain confirmed the participation of this residue in the GNN self-glucosylation and indicated the Tyr198 residue as an additional, although less active, glucosylation site. The physical interaction between GNN and glycogen synthase (GSN) was analyzed by the two-hybrid assay. While the entire GSN was required for full interaction, the C terminus in GNN was more important. Furthermore, mutation in the GNN glucosylation sites did not impair the interaction with GSN. PMID- 15811344 TI - Analysis of the domain properties of the novel cytochrome P450 RhF. AB - The properties of the heme, flavin mononucleotide (FMN) and FeS domains of P450 RhF, from Rhodococcus sp. NCIMB 9784, expressed separately and in combination are analysed. The nucleotide preference, imidazole binding and reduction potentials of the heme and FMN domains are unaltered by their separation. The intact enzyme is monomeric and the flavin is confirmed to be FMN. The two one-electron reduction potentials of the FMN are -240 and -270 mV. The spectroscopic and thermodynamic properties of the FeS domain, masked in the intact enzyme, are revealed for the first time, confirming it as a 2Fe-2S ferredoxin with a reduction potential of -214 mV. PMID- 15811345 TI - Induction of uteroglobin-related protein 2 (Ugrp2) expression by EGF and TGFalpha. AB - Uteroglobin-related protein 2 (UGRP2) is thought to play a role in inflammation and/or epithelial cell differentiation in the lung. Induction of Ugrp2 mRNA expression by epidermal growth factor (EGF) and transforming growth factor alpha was examined using mouse transformed lung Clara cell-derived mtCC cells. The EGF induced increase of Ugrp2 occurred at the transcriptional level that required the EGF receptor and the activation of the ERK-MAPK and phosphoinositide-3 kinase pathways. PMID- 15811346 TI - Polarised distribution of the RNA-binding protein Staufen in differentiated intestinal epithelial cells. AB - mRNA localisation, as a mechanism for directing localised protein synthesis, plays a vital role in the functioning of certain cells, such as neurons and oocytes. Potentially this mechanism may also occur in polarised intestinal epithelial cells. Here we show that Staufen1(55), a protein involved in mRNA localisation and transport, is asymmetrically distributed in differentiated Caco 2 intestinal epithelial cells and partly co-localised with calnexin, a marker of the endoplasmic reticulum. The localisation to the apical region of the cell indicates that Staufen may be involved in localisation of transcripts to this domain. PMID- 15811347 TI - Complementation of two mutant p53: implications for loss of heterozygosity in cancer. AB - Remarkably, a cancer cell rarely possesses two mutant p53 proteins. Instead, mutation of one allele is usually associated with loss of the second p53 allele. Why do not two mutant p53 co-exist? We hypothesize that two different p53 may complement each other, when expressed at equal levels. By titrating trans deficient and DNA-binding-deficient p53 in cells with mutant p53 and by co transfecting distinct mutant p53 in p53-null cells, we demonstrated activation of p53-dependent transcription. We suggest that, due to complementation of two mutant p53, cancer cells need to delete the second p53 allele rather than mutate it. PMID- 15811348 TI - Requirement of the transmembrane semaphorin Sema4C for myogenic differentiation. AB - Semaphorins constitute a large family of signaling proteins that contribute to axonal guidance. Here we demonstrate that the transmembrane semaphorin Sema4C is up-regulated both in the early stage of differentiation of C2C12 mouse skeletal myoblasts into myotubes and during injury-induced muscle regeneration in vivo. Depletion of Sema4C in C2C12 cells resulted in marked attenuation of myotube formation. A fusion protein containing the extracellular Sema domain and a peptide corresponding to the intracellular COOH-terminal region of Sema4C each inhibited the differentiation of C2C12 cells. These findings indicate that Sema4C mediated interaction among myoblasts plays an important role in terminal myogenic differentiation. PMID- 15811349 TI - PICK-1: a scaffold protein that interacts with Nectins and JAMs at cell junctions. AB - Nectin adhesion molecules are involved in the early steps of cell junction formation. Later during the polarisation process, Nectins are components of epithelial adherens junctions where they are indirectly associated with the E cadherin/Catenins complex via the adaptator AF-6. To have a better understanding of Nectin-based cell junctions, we looked for some new Nectins' partners. We demonstrate that the scaffold molecule PICK-1, involved in the clustering of junctional receptors in synaptic junctions, interacts directly with Nectins in a PSD-95/Dlg/ZO-1 domain-dependent manner and is localised at adherens junctions in epithelial cells. Finally, we observed that protein interacting with C-kinase-1 (PICK-1) also interacts directly with the junctional adhesion molecules, and we suggest that PICK-1 could be involved in the regulation of both adherens and tight junctions in epithelial cells. PMID- 15811350 TI - Pi binding by the F1-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane. AB - Pi binding by the F(1)-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane (E. coli F(1)) was examined by two methods: the centrifuge column procedure [Penefsky, H.S. (1977) J. Biol. Chem. 252, 2891-2899] and the Paulus pressure dialysis cell [Paulus, H. (1969) Anal. Biochem. 32, 91-100]. The latter is an equilibrium dialysis-type procedure. Pi binding by beef heart F(1) could be determined by either procedure. However, direct binding of Pi to E. coli F(1) could be determined adequately only in the Paulus cell which indicated more than two binding sites per mol of enzyme with a K(d) in the range of 0.1 mM. It is concluded that previous failure to observe Pi binding to E. coli F(1) with the centrifuge column procedure is due to a rapid rate of dissociation of Pi from the E. coli enzyme which results in loss of Pi during transit of the enzyme-Pi complex through the column. PMID- 15811351 TI - Testosterone, reproductive stage, and territorial behavior of male and female European stonechats Saxicola torquata. AB - We investigated territorial behavior and circulating testosterone (T) levels in a multiple-brooded population of the European stonechat, a socially monogamous passerine bird with biparental care. Between arrival at and departure from the breeding territories, we (1) quantified behavior of both sexes in response to a simulated territorial intrusion (STI) of a male conspecific and (2) measured plasma T concentrations in males and females. Male response scores to a STI and male T concentrations varied with phase, but there was no temporal association between plasma T levels and the intensity of territorial behavior. During both two sexual and two parental phases, at least half of the tested males showed aggressive responses. About 20% of the tested males responded with courtship prior to laying of the first clutch, but none of the males courted during later phases. Age had a positive overall effect on male plasma T. Females also reacted to the STI of a male, but their responses did not vary with breeding phase. Female plasma T varied with phase, being elevated during production of the first but not of the second clutch. As with males, female responses to the STI were not correlated with T levels. Responses of pair partners were positively correlated with each other. We conclude that modulation of male territorial aggression with breeding phase is not regulated by changes of plasma T concentrations. In light of other studies showing reduced male aggression by pharmacological inhibition of cellular actions of T, we propose that T is permissive for male territorial aggression, but does not mediate short-term changes associated with breeding phase. The function of the high female plasma T concentrations during formation of first clutches could be related to the production of eggs with high concentrations of androgens. PMID- 15811352 TI - Effects of testosterone metabolites on copulation, medial preoptic dopamine, and NOS-immunoreactivity in castrated male rats. AB - The medial preoptic area (MPOA) is an important integrative site for male sexual behavior. Dopamine (DA) is released in the MPOA of male rats shortly before and during copulation. In a previous study, we identified 17beta-estradiol (E(2)) as the metabolite of testosterone (T) that maintains MPOA basal extracellular DA levels. However, the presence of dihydrotestosterone (DHT), an androgenic metabolite of T, is required for the female-induced increase in MPOA DA observed during copulation. Recently, we reported that assays of MPOA tissue DA content showed that castrates actually had more stored DA than did gonadally intact males. Therefore, the reduction in extracellular levels in castrates was not due to decreased availability of DA; most likely it was due to decreased release. Furthermore, T upregulates neuronal nitric oxide synthase (nNOS) in the MPOA. NO has been implicated in the regulation of DA release in the MPOA. It is not known, however, which metabolite(s) of T regulate(s) tissue stores of DA and/or nNOS in the MPOA of male rats. The present experiments were designed to test the following: (1) whether E(2), DHT, or the combination of the two influences MPOA DA tissue levels, an indication of stored DA, in male rat castrates; and (2) whether E(2), DHT, or the combination of the two influences NOS-ir in the MPOA of castrated male rats. The results indicate that E(2) up-regulates nNOS-ir in the MPOA and maintains tissue content of DA at levels similar to those in T-treated rats. DHT did not influence nNOS-ir, while attenuating the effect of castration on tissue DA content. PMID- 15811353 TI - Differential effects of 11-ketotestosterone on dimorphic traits in a teleost with alternative male reproductive morphs. AB - Species with dimorphic males typically have one morph (type I) associated with territoriality and courtship, and another morph (type II) associated with cuckoldry. In teleost fishes, type I males generally have higher levels of the androgen 11-ketotestosterone (KT) than type II males. Is KT causal to phenotypic differences between morphs? We investigated this question in the midshipman fish (Porichthys notatus) in which type I males have detectable levels of KT during the breeding season, whereas levels in type II males are usually undetectable. Type I midshipman will either cuckold or hold territories and court females, whereas type II males are only known to cuckold. Acoustic courtship by type I's is supported by the sonic motor nucleus, which innervates a sound-producing sonic muscle. Type I males have larger sonic motor nuclei and larger sonic muscles than type II males, consistent with the more dynamic vocal repertoire of type I's. Here, we tested whether intraperitoneal KT implants in adult type II males would induce type I male-like traits in brain, sonic muscle, and behavior. Type II's treated with KT did not differ from blank-implanted type II's in sonic motor nucleus volume. Sonic muscle mass increased in KT-implanted type II's, but did not reach the relative mass naturally observed in type I's. While neither territoriality nor courtship were induced, cuckoldry behavior intensified in KT implanted type II's. Thus, for some but not all characters, KT exaggerated the expression of already existing type II male traits rather than inducing the type I male-like traits of territoriality, courtship, and an expansive vocal motor system. PMID- 15811354 TI - Stress-induced reductions of sensory reactivity in female rats depend on ovarian hormones and the application of a painful stressor. AB - The current experiments occurred in the context of disputes in the literature concerning whether inescapable stress causes differential changes in sensory reactivity, which could lead to differences in many learning procedures. We tested rats for differences in sensitivity and responsivity to acoustic stimuli through the use of the acoustic startle response (ASR) 2 h after stressor exposure and ambulatory activity 24 h later in the open field. Stressed females showed reduced responsivity to acoustic stimuli with no apparent shift in stimulus sensitivity. Males did not show differences in either reactivity index following stressor exposure. Reduced responsivity did not occur if females had been OVX (OVX alone did not effect stimulus responsivity or sensitivity). All groups that experienced tailshock stress also had reduced open field activity 24 h later. Restraint for 2 h did not reduce stimulus responsivity in the ASR or open field activity in female rats. Acute reductions in ASRs after a painful stressor appear to be a feature specific to females, with an apparent role of ovarian hormones as a modulator of the effect. Possible hormone and/or immunological mechanisms of these sex-specific effects are discussed. Understanding the mechanisms of this stressor-induced reduction in sensory reactivity could advance our knowledge of how individual differences in ovarian hormone levels influence the physical and psychological processes by which females acutely respond and later recover from traumatic events. PMID- 15811355 TI - Oxytocinergic activity is linked to lower blood pressure and vascular resistance during stress in postmenopausal women on estrogen replacement. AB - Estrogen administration results in increased release of the oxytocin (OT) prohormone reflected by increases in oxytocin intermediate peptide (OT Int) in both animal models and humans, and sequential treatment of ovariectomized rats with estrogen/progesterone then progesterone withdrawal leads to increased hypothalamic OT mRNA. Blood pressure (BP) reductions have been related to increased exogenous and endogenous OT in rats and to higher endogenous OT activity in premenopausal women, but not previously in postmenopausal women. Thus, we used plasma obtained at rest and during a speech stressor from 54 postmenopausal women who participated in a 6-month randomized trial of oral conjugated estrogens vs. placebo to examine effects of estrogen replacement therapy (ERT) on plasma OT and OT Int levels and their relationships to changes in BP during the trial. ERT alone and with progesterone (but not placebo) led to significant increases in plasma levels of OT Int, but no change in plasma OT levels. Women showing greater increases in OT Int during treatment showed greater decreases in BP and total vascular resistance during a series of behavioral stressors compared to women with moderate or no increases in OT Int, even after controlling for effects related to treatment condition or to changes in plasma estradiol. The findings suggest that enhanced oxytocinergic activity may contribute to BP decreases associated with ERT in more responsive postmenopausal women. PMID- 15811356 TI - Social stress decreases marking behavior independently of testosterone in Mongolian gerbils. AB - This study examined the endogenous androgen regulation of the marking behavior in Mongolian gerbils (Meriones unguiculatus). In the first experiment, developmental changes of fecal testosterone levels, ventral gland growth, and the marking frequency of male gerbils were investigated. From 9 weeks of age, marking frequency increased with increases in fecal testosterone levels and ventral gland size. The ventral gland size and marking frequency were significantly correlated to the fecal testosterone level. In the second experiment, we hypothesized that reduction in the marking frequency of subordinate males after social confrontations was controlled by a decrease in the circulating testosterone level, and we followed changes in marking frequency, endocrine status, and ventral gland size after social confrontations in which two adult male gerbils established their social ranks by fighting. As expected, marking frequency and ventral gland size were significantly related to social rank, that is, marking frequency was higher among dominant gerbils and lower among subordinates. In addition, fecal corticosterone levels among subordinates were higher than those of dominant animals. However, neither the fecal and plasma testosterone levels, nor testis size, differed between dominant and subordinate gerbils. These results revealed that endogenous androgen played a role in regulating marking behavior and ventral gland size during the developmental stage and that the reductions in marking frequency and ventral gland size occurring in subordinate males after social confrontations were not directly regulated by androgen changes. PMID- 15811357 TI - Sex differences in the long-term effect of preweanling isolation stress on memory retention. AB - Social experiences during development can powerfully modulate later neuroendocrine and behavioral system. In the present study, male and female rat pups experienced daily bouts of social isolation for 6 h per day or control conditions during the third postnatal week. Performance on a 12-arm radial maze with 8 arms consistently baited with food reward was examined in adulthood. During the social isolation, both male and female pups exhibited a significant increase in plasma corticosterone levels. When tested on the radial arm maze as adults, the performance of female rats that had experienced social isolation during development was not affected; however, male rats in the isolation condition initially exhibited impairments in working memory but not reference memory. Despite achieving comparable asymptotic levels of performance on the maze, male rats that experienced social isolation during the third week demonstrated disruption in working memory retention when radial arm maze trials were interrupted after the fourth arm choice. Thus, while male rats that experience social isolation during the third week of life eventually perform comparably to controls on the standard radial arm maze task, their ability to retain information over a delay remains impaired. These findings highlight an important sex difference in the long-term effects of stress during this period of late preweanling development. PMID- 15811358 TI - The hormonal response of female European Stonechats to a territorial intrusion: the role of the male partner. AB - In many bird species, the female participates in defending a pair's breeding territory, however, the endocrine control mechanism of female aggressive behavior is largely unknown. The general statement that androgens are involved in the regulation of aggressive behavior is based on studies conducted only in males. Here, we tested whether paired female stonechats show a hormonal response to a simulated male territorial intruder. Since in males of territorial bird species androgen levels usually increase following a male-male encounter, we measured androgen-levels before and after a simulated male intrusion. In addition, we measured estradiol, the main gonadal hormone in females, and corticosterone, a stress hormone. The results show that a male intruder does not affect any of the measured hormones in females. In a second experiment, we also tested whether the endocrine state of the male partner affects the hormonal response of females to a male intruder by comparing the hormonal response of females paired with pharmacologically castrated males and females paired with control males. Females paired with pharmacologically castrated males had lower corticosterone levels both before and after the intrusion than females paired with control males. Additionally, in both groups, female corticosterone levels were increased following a male intrusion. We suggest that the differences found between females paired with pharmacologically castrated males and females paired with control males are due to differences in intra-pair interactions. PMID- 15811359 TI - Gonadal hormones modulate the display of submissive behavior in socially defeated female Syrian hamsters. AB - There are striking differences in the behavioral response to social defeat between male and female Syrian hamsters. Whereas males exhibit a prolonged behavioral response to defeat (i.e., conditioned defeat), many females remain aggressive or show only a transient submissive response following defeat. The current study tested the hypothesis that sex steroids underlie this differential behavioral responsivity to social defeat. Female hamsters were ovariectomized and implanted with Silastic capsules containing estradiol (E(2)), testosterone (T), progesterone (P), dihydrotestosterone (DHT), or a blank capsule (no hormone replacement). After a 3-week recovery period, each subject was placed inside the home cage of a larger, more aggressive female for four 5-min defeat trials. The following day, each animal was tested for conditioned defeat by testing it in its own home cage in the presence of a smaller, non-aggressive intruder. Submissive, aggressive, social, and nonsocial behaviors were subsequently scored. Hamsters receiving E(2) or T displayed significantly lower levels of submissive behavior than did animals receiving P, DHT, or no hormone replacement. There were no significant differences in aggressive behavior among groups. These data suggest that gonadal hormones can influence submissive behavior in female hamsters. Collectively, these results suggest that the sex differences observed in conditioned defeat may, in part, be explained by sex differences in circulating gonadal hormones. PMID- 15811360 TI - Testosterone and autumn territorial behavior in male red grouse Lagopus lagopus scoticus. AB - In many bird species, males exhibit territorial aggression outside the breeding season, when testosterone concentrations are low and may not regulate territorial behaviors. The hormonal regulation of aggression at this time of year has only been studied in passerine birds. Here, we investigated the role of testosterone in the regulation of aggression in a non-passerine bird, the red grouse Lagopus lagopus scoticus. Male red grouse are aggressive in early spring when breeding starts, in autumn when they establish territories, and sporadically through much of the winter. We first describe seasonal variations in plasma testosterone concentrations and in the size of males' sexual ornaments, their red combs, which relates to aggressiveness. Testosterone concentrations and comb size were correlated. Both increased in autumn to a peak in October, and then increased again in spring, to a greater peak in early April. Secondly, we experimentally investigated the effects of testosterone, and of an anti-androgen (flutamide) used in combination with an aromatase inhibitor (ATD), on autumn territorial behavior. Males were treated with either empty implants, as controls (C-males), testosterone implants (T-males), or with flutamide and ATD implants (FA-males). One month after implanting, both T- and FA-males had higher concentrations of testosterone than C-males. Comb size, aggressive call rate, and response to playbacks of territorial call all significantly increased in T-males. However, the increase in testosterone in FA-males did not increase comb size or aggressive behavior. In the following spring, after the content of implants was used, FA males had significantly lower testosterone than C-males, and had a reduced seasonal increase in comb size. The results suggest that testosterone plays a significant role in regulating red grouse aggressive behavior in autumn. However, the observation that flutamide and ATD treatment did not reduce territorial behavior, suggests that estradiol may also be involved in the regulation of non breeding aggression. PMID- 15811361 TI - Negative effects of elevated testosterone on female fecundity in zebra finches. AB - Although factors influencing androgen deposition in the avian egg and its effects on nestling fitness are recently receiving considerable attention, little is known about the potential costs of high testosterone levels in the females. Our study aimed at determining the effect of injections of testosterone (T) in female zebra finches (Taeniopygia guttata), on clutch size, egg mass, yolk mass, and yolk androgen content. Females were given a single bolus injection of T in a range of doses after laying the first egg. Results show that administration of T negatively affected clutch size; the strength of this effect increased with increasing doses of T. Females injected with the highest testosterone dose showed suppressed oviposition of the third and the fourth eggs. Interestingly, testosterone administration made females produce eggs with relatively large yolks, suggesting that T may mediate the trade-off between number and size of eggs. Testosterone injection resulted in elevated levels of androgen in the eggs, in contrast to control clutches, which showed a decreasing pattern of androgen concentration along the laying sequence. We conclude that high androgen investment in eggs may be limited by physiological requirements of the ovulatory process. PMID- 15811362 TI - Effects of elevated egg corticosterone levels on behavior, growth, and immunity of yellow-legged gull (Larus michahellis) chicks. AB - Eggs of vertebrates contain steroid hormones of maternal origin that may influence offspring performance. Recently, it has been shown that glucocorticoids, which are the main hormones mediating the stress response in vertebrates, are transmitted from the mother to the egg in birds. In addition, mothers with experimentally elevated corticosterone levels lay eggs with larger concentrations of the hormone, which produce slow growing offspring with high activity of the hypothalamo-adrenal axis under acute stress. However, the effects and function of transfer of maternal corticosterone to the eggs are largely unknown. In the present study, we injected corticosterone in freshly laid eggs of yellow-legged gulls (Larus michahellis), thus increasing the concentration of the hormone within its natural range of variation, and analyzed the effect of manipulation on behavioral, morphological, and immune traits of the offspring in the wild. Eggs injected with corticosterone had similar hatching success to controls, but hatched later. Mass loss during incubation was greater for corticosterone-treated eggs, except for the last laid ones. Corticosterone injection reduced rate and loudness of late embryonic vocalizations and the intensity of chick begging display. Tonic immobility response, reflecting innate fearfulness, was unaffected by hormone treatment. Elevated egg corticosterone concentrations depressed T-cell-mediated immunity but had no detectable effects on humoral immune response to a novel antigen, viability at day 10, or growth. Present results suggest that egg corticosterone can affect the behavior and immunity of offspring in birds and disclose a mechanism mediating early maternal effects whereby stress experienced by females may negatively translate to offspring phenotypic quality. PMID- 15811363 TI - The adrenocortical response of tufted puffin chicks to nutritional deficits. AB - In several seabirds, nutritional state of a nest-bound chick is negatively correlated with the activity of its hypothalamus-pituitary-adrenal (HPA) axis. Increased corticosterone (cort) secretion has been shown to facilitate changes in behavior that allow hungry chicks to obtain more food from parents. However, if parents are not willing/able to buffer their young from temporary food shortages, increased cort secretion could be detrimental to undernourished chicks. In a system where parents are insensitive to chick demands, low benefits and high costs of activation of the HPA-axis in hungry chicks should lead to a disassociation of the nutritional state of the young and the activity of its HPA axis. We tested this novel hypothesis for the tufted puffin (Fratercula cirrhata), a seabird with intermittent provisioning of a nest-bound semi precocial chick. We examined the HPA-axis activity of captive chicks exposed to the following: (1) a short-term (24 h) food deprivation; and (2) an array of prolonged (3 weeks) restrictions in feeding regimens. We found that in response to a short-term food deprivation chicks decreased baseline levels of cort and thyroid hormones. In response to prolonged restrictions, food-limited chicks exhibited signs of nutritional deficit: they had lower body mass, endogenous lipid reserves, and thyroid hormone titers compared to chicks fed ad libitum. However, baseline and maximum acute stress-induced levels of cort were also lower in food-restricted chicks compared to those of chicks fed ad libitum. These results support a major prediction of the study hypothesis that puffin chicks suppress HPA-axis activity in response to short- and long-term nutritional deficits. This physiological adaptation may allow a chick to extend its development in the nest, while eluding detrimental effects of chronic cort elevation. PMID- 15811364 TI - Pair-housing of male and female rats during chronic stress exposure results in gender-specific behavioral responses. AB - Social support has a positive influence on the course of a depression and social housing of rats could provide an animal model for studying the neurobiological mechanisms of social support. Male and female rats were subjected to chronic footshock stress for 3 weeks and pair-housing of rats was used to mimic social support. Rats were isolated or housed with a partner of the opposite sex. A plastic tube was placed in each cage and subsequently used as a 'safe' area in an open field test. Time spent in the tube was used as a measurement of anxiety levels. Chronic stress increased adrenal weights in all groups, except for isolated females who showed adrenal hypertrophy in control conditions. In isolated males, chronic stress resulted in an increase in the time the animals spent in the tube. While stress did not affect this parameter in socially housed males, males with a stressed partner showed a similar response as isolated stressed males. Even though adrenal weights showed that isolated females were more affected by stress, after chronic stress exposure, they spent less time in the tube than socially housed females. Socially housed stressed females spent less time in the 'safe' tube compared to control counterparts, indicating that stress has a gender-specific behavioral effect. IN CONCLUSION: pair-housing had a stress-reducing effect on behavior in males. Isolation of females was stressful by itself. Pair housing of females was not able to prevent stress-induced behavioral changes completely, but appeared to reduce the effects of chronic stress. PMID- 15811365 TI - The parallel superpleated beta-structure as a model for amyloid fibrils of human amylin. AB - Human amylin is a 37 amino acid residue peptide hormone whose fibrillogenesis has been correlated with type 2 diabetes. These fibrils are rope-like bundles of several 5nm diameter protofilaments. Here, we propose, as a model for the protofilament, a variant of the parallel superpleated beta-structure previously derived for amyloid filaments of the yeast prion Ure2p. In the amylin model, individual polypeptides from residues 9 to 37 have a planar S-shaped fold with three beta-strands. These serpentines are stacked in register, with a 0.47 nm axial rise and a small rotational twist per step, generating an array of three parallel beta-sheets in cross-beta conformation. The interior, the two "bays" sandwiched between adjacent sheets, are occupied by non-polar and by polar/uncharged residues that are predicted to form H-bonded ladders, similar to those found in beta-helical proteins. The N-terminal peptide containing a disulfide bond occupies an extraneous peripheral position in the protofilament. The left-handed twist of the beta-sheets is shown to underlie left-handed coiling of amylin protofilaments in fibrils. The model is consistent with current biophysical, biochemical and genetic data and, in particular, affords a plausible explanation for why rodent amylin does not form fibrils. PMID- 15811366 TI - Solution structure of the major DNA-binding domain of Arabidopsis thaliana ethylene-insensitive3-like3. AB - Ethylene-insensitive3 (EIN3) and EIN3-like (EIL) proteins are essential transcription factors in the ethylene signaling of higher plants. The EIN3/EIL proteins bind to the promoter regions of the downstream genes and regulate their expression. The location of the DNA-binding domain (DBD) in the primary structure was unclear, since the proteins show no sequence similarity to other known DBDs. Here, we identify the major DBD of an EIN3/EIL protein, Arabidopsis thaliana EIL3, containing a key mutational site for DNA binding and signaling (ein3-3 site), and determine its solution structure by NMR spectroscopy. The structure consists of five alpha-helices, possessing a novel fold dissimilar to known DBD structures. By a chemical-shift perturbation analysis, a region including the ein3-3 site is suggested to be involved in DNA binding. PMID- 15811368 TI - RNA binding and phosphorylation determine the intracellular distribution of nuclear factors 90 and 110. AB - Members of the nuclear factor 90 (NF90) family of human double-stranded RNA (dsRNA) binding proteins are phosphorylated and translocate into the cytoplasm with the onset of mitosis. We investigated the mechanism of translocation for NF90 and NF110, its larger splice variant. During interphase, NF90 is predominantly nuclear, NF110 is exclusively nuclear, and both are bound to RNA. About half of the NF90 is tethered in the nucleus by RNA bound to the protein's dsRNA-binding motifs. The nuclear localization of NF110 is also dependent on RNA binding but is independent of these motifs, and is governed by contacts made to the protein's unique C terminus. During mitosis, about half of the cytoplasmic NF90 becomes dissociated from RNA, but phosphorylation does not impair the binding affinity of either NF90 or NF110 for dsRNA. We conclude that NF90 and NF110 engage RNA differentially and translocate from the nucleus to the cytoplasm in mitosis because phosphorylation disturbs their interactions with other nuclear proteins. PMID- 15811367 TI - Solution structure and backbone dynamics of the KH-QUA2 region of the Xenopus STAR/GSG quaking protein. AB - The Quaking protein belongs to the family of STAR/GSG domain RNA-binding proteins and is involved in multiple cell signalling and developmental processes in vertebrates, including the formation of myelin. Heteronuclear NMR methods were used to determine the solution structure of a 134 residue fragment spanning the KH and QUA2 homology regions of the Quaking protein from Xenopus laevis (pXqua) in the absence of RNA. The protein is shown to adopt an extended type I KH domain fold that is connected to a structured alpha-helix in the C-terminal QUA2 region by means of a highly flexible linker. A comparison with the solution structure of the related protein splicing factor 1 (SF1) indicates that most aspects of the RNA-binding interface are conserved in pXqua, although the "variable loop" region that follows the second beta-strand possesses two additional alpha-helices. The structure of pXqua provides an appropriate template for building models of important homologues, such as GLD-1 and Sam68. Measurements of the (15)N relaxation parameters of pXqua confirm that the polypeptide backbone of the QUA2 region is more dynamic than that of the KH portion, and that the C-terminal helix is partially structured in the absence of RNA. By comparison with a random coil reference state, the nascent structure in the QUA2 region is estimated to contribute 15.5kJmol(-1) to the change in conformational free energy that occurs on forming a complex with RNA. Since STAR/GSG proteins may regulate alternative splicing by competing with SF1 in the nucleus for specific branch-point sequences that signal intronic RNA, the formation of secondary structure in the QUA2 region in the unbound state of pXqua has important functional consequences. PMID- 15811369 TI - Crystal structure of the RNA 2'-phosphotransferase from Aeropyrum pernix K1. AB - In the final step of tRNA splicing, the 2'-phosphotransferase catalyzes the transfer of the extra 2'-phosphate from the precursor-ligated tRNA to NAD. We have determined the crystal structure of the 2'-phosphotransferase protein from Aeropyrum pernix K1 at 2.8 Angstroms resolution. The structure of the 2' phosphotransferase contains two globular domains (N and C-domains), which form a cleft in the center. The N-domain has the winged helix motif, a subfamily of the helix-turn-helix family, which is shared by many DNA-binding proteins. The C domain of the 2'-phosphotransferase superimposes well on the NAD-binding fold of bacterial (diphtheria) toxins, which catalyze the transfer of ADP ribose from NAD to target proteins, indicating that the mode of NAD binding by the 2' phosphotransferase could be similar to that of the bacterial toxins. The conserved basic residues are assembled at the periphery of the cleft and could participate in the enzyme contact with the sugar-phosphate backbones of tRNA. The modes by which the two functional domains recognize the two different substrates are clarified by the present crystal structure of the 2'-phosphotransferase. PMID- 15811370 TI - Thermodynamic and kinetic basis for the relaxed DNA sequence specificity of "promiscuous" mutant EcoRI endonucleases. AB - Promiscuous mutant EcoRI endonucleases produce lethal to sublethal effects because they cleave Escherichia coli DNA despite the presence of the EcoRI methylase. Three promiscuous mutant forms, Ala138Thr, Glu192Lys and His114Tyr, have been characterized with respect to their binding affinities and first-order cleavage rate constants towards the three classes of DNA sites: specific, miscognate (EcoRI*) and non-specific. We have made the unanticipated and counterintuitive observations that the mutant restriction endonucleases that exhibit relaxed specificity in vivo nevertheless bind more tightly than the wild type enzyme to the specific recognition sequence in vitro, and show even greater preference for binding to the cognate GAATTC site over miscognate sites. Binding preference for EcoRI* over non-specific DNA is also improved. The first-order cleavage rate constants of the mutant enzymes are normal for the cognate site GAATTC, but are greater than those of the wild-type enzyme at EcoRI* sites. Thus, the mutant enzymes use two mechanisms to partially bypass the multiple fail-safe mechanisms that protect against cleavage of genomic DNA in cells carrying the wild-type EcoRI restriction-modification system: (a) binding to EcoRI* sites is more probable than for wild-type enzyme because non-specific DNA is less effective as a competitive inhibitor; (b) the combination of increased affinity and elevated cleavage rate constants at EcoRI* sites makes double-strand cleavage of these sites a more probable outcome than it is for the wild-type enzyme. Semi quantitative estimates of rates of EcoRI* site cleavage in vivo, predicted using the binding and cleavage constants measured in vitro, are in accord with the observed lethal phenotypes associated with the three mutations. PMID- 15811371 TI - Formation and maintenance of tubular membrane projections require mechanical force, but their elongation and shortening do not require additional force. AB - Living cells develop their own characteristic shapes depending on their physiological functions, and their morphologies are based on the mechanical characteristics of the cytoskeleton and of membranes. To investigate the role of lipid membranes in morphogenesis, we constructed a simple system that can manipulate liposomes and measure the forces required to transform their shapes. Two polystyrene beads (1 microm in diameter) were encapsulated in giant liposomes and were manipulated using double-beam laser tweezers. Without any specific interaction between the lipid membrane and beads, mechanical forces could be applied to the liposome membrane from the inside. Spherical liposomes transformed into a lemon shape with increasing tension, and tubular membrane projections were subsequently generated in the tips at either end. This process is similar to the liposomal transformation caused by elongation of encapsulated cytoskeletons. In the elongation stage of lemon-shaped liposomes, the force required for the transformation became larger as the end-to-end length increased. Just before the tubular membrane was generated, the force reached the maximum strength (approximately 11 pN). However, immediately after the tubular membrane developed, the force suddenly decreased and was maintained at a constant strength (approximately 4 pN) that was independent of further tube elongation or shortening, even though there was no excess membrane reservoir as occurs in living cells. When the tube length was shortened to approximately 2 microm, the liposome reversed to a lemon shape and the force temporarily increased (to approximately 7 pN). These results indicate that the simple application of mechanical force is sufficient to form a protrusion in a membrane, that a critical force and length is needed to form and to maintain the protrusion, and suggest that the lipid bilayer itself has the ability to buffer the membrane tension. PMID- 15811372 TI - The native energy landscape for interleukin-1beta. Modulation of the population ensemble through native-state topology. AB - A minimalist Go-model, with no energetic frustration in the native conformation, has been shown to describe accurately the folding pathway of the beta-trefoil protein, interleukin-1beta (IL-1beta). While it appears that these models successfully model transition states and intermediates between the unfolded and native ensembles, it is unclear how accurately they capture smaller, yet biologically relevant, structural changes within the native ensemble after energetic perturbation. Here, we address the following questions. Can a simple Go model of interleukin-1beta, based on native topology, describe changes in structural properties of the native ensemble as the protein stability is changed? Or is it necessary to include a more explicit representation of atoms, electrostatic, hydrogen bonding, and van der Waals forces to describe these changes? The native ensemble of IL-1beta was characterized using a variety of experimental probes under native (0 M NaCl, guanidine hydrochloride (Gdn-HCl)), moderately destabilized (0 M NaCl, 0.8 M Gdn-HCl), and in moderate salt concentration (0.8 M NaCl, 0 M Gdn-HCl). Heteronuclear (1)H-(15)N nuclear Overhauser effect spectroscopy (NOESY) and heteronuclear single quantum correlation (HSQC) NMR spectra confirmed that the beta-trefoil global fold was largely intact under these three conditions. However, 25 of the 153 residues throughout the chain did demonstrate (13)C and (1)H-(15)N chemical shifts when perturbed with 0.8 M NaCl or Gdn-HCl. Despite large differences in protection factors from solvent hydrogen-deuterium exchange for all residues between stable (0 M Gdn-HCl) and destabilized (0.8 M Gdn-HCl) IL-1beta, no difference in steady state (15)N-(1)H NOE enhancements were measured. Thus, the chemical shifts correlate with a global but limited increase in residue flexibility in the presence of Gdn-HCl. Minimalist simulations highlight the regions of greatest position shift between native and 0.8 M Gdn-HCl, which were determined experimentally. This correlation demonstrates that structural changes within the native ensemble of IL-1beta are, at least partially, governed by the principle of minimal energetic frustration. PMID- 15811373 TI - Atomic resolution structures of CTX-M beta-lactamases: extended spectrum activities from increased mobility and decreased stability. AB - Extended spectrum beta-lactamases (ESBLs) confer bacterial resistance to third generation cephalosporins, such as cefotaxime and ceftazidime, increasing hospital mortality rates. Whereas these antibiotics are almost impervious to classic beta-lactamases, such as TEM-1, ESBLs have one to four orders greater activity against them. The origins of this activity have been widely studied for the TEM and SHV-type ESBLs, but have received less attention for the CTX-M beta lactamases, an emerging family that is now the dominant ESBL in several regions. To understand how CTX-M beta-lactamases achieve their remarkable activity, biophysical and structural studies were undertaken. Using reversible, two-state thermal denaturation, it was found that as these enzymes evolve a broader substrate range, they sacrifice stability. Thus, the mutant enzyme CTX-M-16 is eightfold more active against ceftazidime than the pseudo-wild-type CTX-M-14 but is 1.9 kcal/mol less stable. This is consistent with a "stability-activity tradeoff," similar to that observed in the evolution of other resistance enzymes. To investigate the structural basis of enzyme activity and stability, the structures of four CTX-M enzymes were determined by X-ray crystallography. The structures of CTX-M-14, CTX-M-27, CTX-M-9 and CTX-M-16 were determined to 1.10 Angstroms, 1.20 Angstroms, 0.98 Angstroms and 1.74 Angstroms resolution, respectively. The enzyme active sites resemble those of the narrow-spectrum TEM-1 and SHV-1, and not the enlarged sites typical of ESBL mutants such as TEM-52 and TEM-64. Instead, point substitutions leading to specific interactions may be responsible for the improved activity against ceftazidime and cefotaxime, consistent with observations first made for the related Toho-1 enzyme. The broadened substrate range of CTX-M-16 may result from coupled defects in the enzyme's B3 strand, which lines the active site. Substitutions Val231-->Ala and Asp240-->Gly, which convert CTX-M-14 into CTX-M-16, occur at either end of this strand. These defects appear to increase the mobility of B3 based on anisotropic B-factor analyses at ultrahigh resolution, consistent with stability loss and activity gain. The unusually high resolution of these structures that makes such analyses possible also makes them good templates for inhibitor discovery. PMID- 15811374 TI - Crystal structure of the mosquito-larvicidal toxin Cry4Ba and its biological implications. AB - Cry4Ba, isolated from Bacillus thuringiensis subsp. israelensis, is specifically toxic to the larvae of Aedes and Anopheles mosquitoes. The structure of activated Cry4Ba toxin has been determined by multiple isomorphous replacement with anomalous scattering and refined to R(cryst) = 20.5% and R(free)= 21.8% at 1.75 Angstroms resolution. It resembles previously reported Cry toxin structures but shows the following distinctions. In domain I the helix bundle contains only the long and amphipathic helices alpha3-alpha7. The N-terminal helices alpha1 alpha2b, absent due to proteolysis during crystallisation, appear inessential to toxicity. In domain II the beta-sheet prism presents short apical loops without the beta-ribbon extension of inner strands, thus placing the receptor combining sites close to the sheets. In domain III the beta-sandwich contains a helical extension from the C-terminal strand beta23, which interacts with a beta-hairpin excursion from the edge of the outer sheet. The structure provides a rational explanation of recent mutagenesis and biophysical data on this toxin. Furthermore, added to earlier structures from the Cry toxin family, Cry4Ba completes a minimal structural database covering the Coleoptera, Lepidoptera, Diptera and Lepidoptera/Diptera specificity classes. A multiple structure alignment found that the Diptera-specific Cry4Ba is structurally more closely similar to the Lepidoptera-specific Cry1Aa than the Coleoptera-specific Cry3Aa, but most distantly related to Lepidoptera/Diptera-specific Cry2Aa. The structures are most divergent in domain II, supporting the suggestion that this domain has a major role in specificity determination. They are most similar in the alpha3 alpha7 major fragment of domain I, which contains the alpha4-alpha5 hairpin crucial to pore formation. The collective knowledge of Cry toxin structure and mutagenesis data will lead to a more critical understanding of the structural basis for receptor binding and pore formation, as well as allowing the scope of diversity to be better appreciated. PMID- 15811375 TI - Nuclear magnetic resonance characterization of the refolding intermediate of beta2-microglobulin trapped by non-native prolyl peptide bond. AB - beta(2)-Microglobulin (beta2-m), a light chain of the major histocompatibility complex type I, is also found as a major component of amyloid fibrils formed in dialysis-related amyloidosis. Denaturation of beta2-m is considered to initiate the formation of fibrils. To clarify the mechanism of fibril formation, it is important to characterize the intermediate conformational states at the atomic level. Here, we investigated the refolding of beta2-m from the acid-unfolded state by heteronuclear magnetic resonance and circular dichroism spectroscopies. At low temperature, beta2-m refolded slowly, accumulating a rate-limiting intermediate with non-native chemical shift dispersions for several residues, but with compactness and secondary structures similar to those of the native protein. beta2-m has a cis proline residue at Pro32, located on the turn connecting the betaB and betaC strands. The slow refolding phase disappeared upon mutation of Pro32 to Val, indicating that Pro32 is responsible for the accumulation of the intermediate. The distribution of the perturbed residues in the intermediate suggests that the non-native prolyl peptide bond of Pro32 affects large areas of the molecule. A cis proline residue is common to various immunoglobulin domains involved in amyloidosis, implying that a non-native prolyl peptide bond that might occur under physiological conditions is related to the amyloidogenicity of these immunoglobulin domains. PMID- 15811376 TI - Structural basis for APPTPPPLPP peptide recognition by the FBP11WW1 domain. AB - WW domains are small protein-protein interaction modules that recognize proline rich stretches in proteins. The class II tandem WW domains of the formin binding protein 11 (FBP11) recognize specifically proteins containing PPLPp motifs as present in the formins that are involved in limb and kidney development, and in the methyl-CpG-binding protein 2 (MeCP2), associated with the Rett syndrome. The interaction involves the specific recognition of a leucine side-chain. Here, we report on the novel structure of the complex formed by the FPB11WW1 domain and the formin fragment APPTPPPLPP revealing the specificity determinants of class II WW domains. PMID- 15811377 TI - Dynamic mechanism for the serpin loop insertion as revealed by quantitative kinetics. AB - The serpin conformational change by insertion of the reactive center loop into beta-sheet A plays a central role in multiple physiological consequences such as serine proteinase inhibition, latency and serpinopathic polymerization. To study the dynamic mechanism for the loop insertion, a novel kinetic method was established utilizing the ovalbumin mutant R339T/A352R; the loop insertion progressed after the cleavage of P1-P1' (Arg352-Ser353) by trypsin was quenched at pH 8 and 0.5 degrees C, and different conformers were quantified by separation using ion-exchange HPLC. The apparent first-order rate constant k(app) determined for various R339T/A352R derivatives differing in conformational stability was greatly increased by lowering the pH. The pH-dependence of k(app) indicated that the protonation of side-chain(s) with a pK(a) value of around 4.6 is a pre requisite for the loop insertion. The theoretical rate constant k for the protonated form calculated from k(app) was highly variable, depending on the ovalbumin derivative; structural modifications that give increased mobility to helix F and the sheet-A half (s3A/s2A/s1A) resulted in a striking increase in the loop insertion rate constant k. The k values were determined at different temperatures for all the ovalbumin derivatives, and DeltaH(double dagger) and DeltaS(double dagger) values for the loop insertion reaction were determined according to the transition theory. The formation of the transition state was highly endothermic with minor entropy gain, requiring a DeltaG(double dagger) larger than 18 kcal/mol, which can offset the hydrogen-bond cleavages between s3A and s5A. These results are consistent with the transition state with an opened sheet A and altered orientation of helix F. PMID- 15811378 TI - Electrostatic interactions contribute to reduced heat capacity change of unfolding in a thermophilic ribosomal protein l30e. AB - The origin of reduced heat capacity change of unfolding (DeltaC(p)) commonly observed in thermophilic proteins is controversial. The established theory that DeltaC(p) is correlated with change of solvent-accessible surface area cannot account for the large differences in DeltaC(p) observed for thermophilic and mesophilic homologous proteins, which are very similar in structures. We have determined the protein stability curves, which describe the temperature dependency of the free energy change of unfolding, for a thermophilic ribosomal protein L30e from Thermococcus celer, and its mesophilic homologue from yeast. Values of DeltaC(p), obtained by fitting the free energy change of unfolding to the Gibbs-Helmholtz equation, were 5.3 kJ mol(-1) K(-1) and 10.5 kJ mol(-1) K(-1) for T.celer and yeast L30e, respectively. We have created six charge-to-neutral mutants of T.celer L30e. Removal of charges at Glu6, Lys9, and Arg92 decreased the melting temperatures of T.celer L30e by approximately 3-9 degrees C, and the differences in melting temperatures were smaller with increasing concentration of salt. These results suggest that these mutations destabilize T.celer L30e by disrupting favorable electrostatic interactions. To determine whether electrostatic interactions contribute to the reduced DeltaC(p) of the thermophilic protein, we have determined DeltaC(p) for wild-type and mutant T.celer L30e by Gibbs-Helmholtz and by van't Hoff analyses. A concomitant increase in DeltaC(p) was observed for those charge-to-neutral mutants that destabilize T.celer L30e by removing favorable electrostatic interactions. The crystal structures of K9A, E90A, and R92A, were determined, and no structural change was observed. Taken together, our results support the conclusion that electrostatic interactions contribute to the reduced DeltaC(p) of T.celer L30e. PMID- 15811379 TI - Crystal structure of cockroach allergen Bla g 2, an unusual zinc binding aspartic protease with a novel mode of self-inhibition. AB - The crystal structure of Bla g 2 was solved in order to investigate the structural basis for the allergenic properties of this unusual protein. This is the first structure of an aspartic protease in which conserved glycine residues, in two canonical DTG triads, are substituted by different amino acid residues. Another unprecedented feature revealed by the structure is the single phenylalanine residue insertion on the tip of the flap, with the side-chain occupying the S1 binding pocket. This and other important amino acid substitutions in the active site region of Bla g 2 modify the interactions in the vicinity of the catalytic aspartate residues, increasing the distance between them to approximately 4A and establishing unique direct contacts between the flap and the catalytic residues. We attribute the absence of substantial catalytic activity in Bla g 2 to these unusual features of the active site. Five disulfide bridges and a Zn-binding site confer stability to the protein, which may contribute to sensitization at lower levels of exposure than other allergens. PMID- 15811380 TI - Architecture of the ribosome-channel complex derived from native membranes. AB - The mammalian Sec61 complex forms a protein translocation channel whose function depends upon its interaction with the ribosome and with membrane proteins of the endoplasmic reticulum (ER). To study these interactions, we determined structures of "native" ribosome-channel complexes derived from ER membranes. We find that the ribosome is linked to the channel by seven connections, but the junction may still provide a path for domains of nascent membrane proteins to move into the cytoplasm. In addition, the native channel is significantly larger than a channel formed by the Sec61 complex, due to the presence of a second membrane protein. We identified this component as TRAP, the translocon-associated protein complex. TRAP interacts with Sec61 through its transmembrane domain and has a prominent lumenal domain. The presence of TRAP in the native channel indicates that it may play a general role in translocation. Crystal structures of two Sec61 homologues were used to model the channel. This analysis indicates that there are four Sec61 complexes and two TRAP molecules in each native channel. Thus, we suggest that a single Sec61 complex may form a conduit for translocating polypeptides, while three copies of Sec61 play a structural role or recruit accessory factors such as TRAP. PMID- 15811381 TI - Conversion of the tetrameric restriction endonuclease Bse634I into a dimer: oligomeric structure-stability-function correlations. AB - The Bse634I restriction endonuclease is a tetramer and belongs to the type IIF subtype of restriction enzymes. It requires two recognition sites for its optimal activity and cleaves plasmid DNA with two sites much faster than a single-site DNA. We show that disruption of the tetramerisation interface of Bse634I by site directed mutagenesis converts the tetrameric enzyme into a dimer. Dimeric W228A mutant cleaves plasmid DNA containing one or two sites with the same efficiency as the tetramer cleaves the two-site plasmid. Hence, the catalytic activity of the Bse634I tetramer on a single-site DNA is down-regulated due to the cross talking interactions between the individual dimers. The autoinhibition within the Bse634I tetramer is relieved by bridging two DNA copies into the synaptic complex that promotes fast and concerted cleavage at both sites. Cleavage analysis of the oligonucleotide attached to the solid support revealed that Bse634I is able to form catalytically competent synaptic complexes by bridging two molecules of the cognate DNA, cognate DNA-miscognate DNA and cognate DNA-product DNA. Taken together, our data demonstrate that a single W228A mutation converts a tetrameric type IIF restriction enzyme Bse634I into the orthodox dimeric type IIP restriction endonuclease. However, the stability of the dimer towards chemical denaturants, thermal inactivation and proteolytic degradation are compromised. PMID- 15811382 TI - Asymmetric binding between SecA and SecB two symmetric proteins: implications for function in export. AB - SecB, a small tetrameric chaperone in Escherichia coli, facilitates export of precursor polypeptides from the cytoplasm to the periplasmic space. During this process, SecB displays two modes of binding. As a chaperone, it binds promiscuously to precursors to maintain them in a non-native conformation. SecB also demonstrates specific recognition of, and binding to, SecA. SecB with the precursor tightly bound enters an export-active complex with SecA and must pass the ligand to SecA at the translocon in the membrane. Here we use variants of SecA and SecB to further probe these interactions. We show that, unexpectedly, the binding between the two symmetric molecules is asymmetric and that the C terminal alpha-helices of SecB bind in the interfacial region of the SecA dimer. We suggest that disruption of this interface by SecB facilitates conformational changes of SecA that are crucial to the transfer of the precursor from SecB to SecA. PMID- 15811383 TI - Caveolin-1 interacts directly with dynamin-2. AB - Caveolin is the principal component of caveolae in vivo. In addition to a structural role, it is believed to play a scaffolding function to organize and inactivate signaling molecules that are concentrated on the cytoplasmic surface of caveolar membranes. The large GTPase dynamin has been shown to mediate the scission of caveolae from the plasma membrane, although it is unclear if dynamin interacts directly with caveolin or via accessory proteins. Therefore, the goal of this study was to test whether dynamin associates with caveolae via a direct binding to the caveolin 1 (Cav1) protein. Immunoelectron microscopy of lung endothelium or a cultured hepatocyte cell line stained with antibodies for Dyn2 and Cav1 shows that these proteins co-localize to caveolae. To further define this interaction biochemically, in vitro experiments were performed using glutathione-S-transferase (GST)-Dyn2 and GST-Cav1 fusion proteins, which demonstrated a direct interaction between these proteins. This interaction appears to be mediated by the proline-arginine-rich domain (PRD) of Dyn2, as a GST-PRD fragment binds Cav1 while GST-Dyn2DeltaPRD does not. Further, in vitro binding studies using two Dyn2 spliced forms and Cav1 peptides immobilized on paper identify specific domains of Cav1 that bind Dyn2. Interestingly, these Cav1 binding domains differ markedly between two spliced variant forms of Dyn2. In support of these distinctive physical interactions, we find that the different Dyn2 forms, when expressed as GTPase-defective mutants, exert markedly different inhibitory effects on caveolae internalization, as assayed by cholera toxin uptake. These studies provide the first evidence for a direct interaction between dynamin and the caveolin coat, and demonstrate a selectivity of one Dyn2 form toward the caveolae-mediated endocytosis. PMID- 15811384 TI - Genetic analysis of learning and memory deficits in senescence-accelerated mouse (SAM). AB - Genetic analysis of learning and memory deficits (LMD) in senescence-accelerated mouse P8 (SAMP8) was performed by cross-mating SAMP8 and Japanese Fancy Mouse 1 (JF1). The incidence of LMD in the F2 generation showed a 3:1 segregation ratio of mice with LMD to normal mice, and the incidence of LMD in the backcross generation of the F1 to JF1 parental strain was in agreement with a 1:1 ratio of mice with LMD to normal mice. Estimation of the number of genes involved in the development of LMD using Wright's formula showed that at least two to four genes are involved. These results suggest that the inheritance of LMD is polygenically controlled and that there may be a single major gene, but this locus is not sex linked. Moreover, hormonal influence on the development of LMD in SAMP8 females is of a genotype-dependent manner. PMID- 15811385 TI - Social instability in female rats: the relationship between stress-related and anxiety-like consequences. AB - It is generally believed that anxiety and depression develop in response to stressful events that chronically increase glucocorticoid production (which in turn affects various neurotransmitter systems). In contrast to depression, however, the relationship between chronic stress and anxiety is less clear, as anxiety patients often show normal glucocorticoid levels and respond normally to dexamethasone challenge. Here we report on the interaction between symptoms of chronic stress and anxiety in female rats by making use of the social instability model of anxiety. Subjects were exposed to alternating 24 h periods of isolation and moderate crowding for 14 days. Symptoms of chronic stress and anxiety-like behaviour in the social interaction test were evaluated on the 15th day, i.e. 1 day after the last crowding phase. Social instability resulted in decreased weight gain, and chronically elevated plasma glucocorticoid levels only in rats that showed high aggressiveness during the crowding phases. In contrast, anxiety like behaviour was increased irrespective to crowding-related aggressiveness. Thus, the development of chronic stress symptoms and anxiety-like behaviour dissociated: the former was bound to crowding-induced aggressiveness (i.e. to a higher stress load), whereas the latter occurred also when such aggressiveness was low, and symptoms of chronic stress did not develop. This finding is consistent with human data, and suggests that stressful events lead to anxiety in both cases: when stressors do or do not lead to chronic stress responses. Studying the distinctive features of anxieties associated or not with chronic endocrine stress responses would enhance our understanding of this disorder. PMID- 15811386 TI - Potential energetic implications of emesis in the house musk shrew (Suncus murinus). AB - During the course of studies investigating novel anti-emetic therapies we serendipitously observed a previously unreported behaviour related to emesis in the house musk shrew. This behaviour consisted of spontaneous ingestion of vomit in about half of the animals (males and females) in which emesis was induced by either nicotine (4 mg kg-1 sc.) or horizontal motion (1 Hz, 4 cm, 10 min). Analysis of vomit samples and gastric contents revealed that in a "typical" individual the gastric contents would be voided by as few as 3 vomits. Energetic calculations of the metabolisable energy of food, gastric contents, vomit and field metabolic rate (FMR) predict that a male weighing 60 g would lose 17.3% of its hourly energy requirement for FMR if it vomited once. A 40 g female, however, would experience an hourly energy loss of approximately 22.8%. The possible energetic consequences and resulting ecological implications of this unusual behaviour are discussed. PMID- 15811387 TI - Assessment of recovery in the hemiparkinson rat: drug-induced rotation is inadequate. AB - Recovery from apomorphine-induced rotational behavior was compared to sensorimotor and motor function in hemiparkinsonian rats receiving intrastriatal grafts of astrocytes expressing recombinant tyrosine hydroxylase (TH) or control beta-galactosidase (beta-gal). Rats received unilateral intranigral infusions of 6-hydroxydopamine (6-OHDA). Animals with large lesions, as determined by apomorphine-induced rotation, received grafts of astrocytes into the denervated striatum. Behavioral recovery was assessed on days 14-16 post-transplantation using apomorphine-induced rotation, somatosensory neglect, and reaching for pellets using the Montoya staircase method. Rats that received transplants of TH transfected astrocytes showed a 34% decrease in rotational behavior, but no consistent recovery of somatosensory neglect or skilled reaching. Post-mortem histological analyses revealed survival of grafted astrocytes in host striatum and expression of TH at 17 days post-transplantation. We suggest that TH expressing astrocytes may reverse post-synaptic dopamine (DA) receptor supersensitivity; however, sensorimotor and motor abilities are not restored due to a failure by TH-expressing astrocytes to reestablish dopaminergic circuitry. The present results demonstrate the need to utilize a variety of sensory and motor behavioral tests that cohesively provide greater interpretability than a single behavioral measure used in isolation, such as drug-induced rotational behavior, to assess the efficacy of experimental gene therapies. PMID- 15811388 TI - Effects of ethanol intake and ethanol withdrawal on free-running circadian activity rhythms in rats. AB - Chronic alcohol intake and alcohol withdrawal are associated with dramatic disruptions of daily (circadian) biological rhythms in both human alcoholics and experimental animals. The extent to which these observations are due to pharmacological effects on the underlying circadian pacemaker is not known, however, since no human studies and very few animal studies have been conducted under free-running conditions. In the present study, free-running circadian activity (wheel-running) rhythms of rats were monitored before, during and after exposure to either 10% or 20% ethanol solution as the only drinking fluid. Across individuals, both lengthening and shortening of free-running period were observed during ethanol intake, and treatment termination led to either a return to baseline or to an exacerbation of the original ethanol effect. These variable effects appeared to be related to both ethanol concentration and to individual differences in baseline period, such that relatively short free-running period during baseline was associated with greater period-lengthening during ethanol exposure. These bidirectional affects of ethanol on free-running period are generally similar to effects seen previously with other psychoactive drugs, including antidepressants. The results of this study indicate that ethanol influences the circadian pacemaker, and that the chronobiological disruptions seen in human alcoholics may be due, in part, to alterations in circadian pacemaker regulation. PMID- 15811389 TI - Characterization of anxiety and habituation profile following global ischemia in rats. AB - The purpose of the current study was to document the behavioral profile of ischemic rats in novel tasks including the elevated plus maze (EPM), the Vogel/conflict model of anxiety and novelty-induced feeding suppression paradigm as well as to further characterize using behavioral monitoring, the response of ischemic animals in existing paradigms such as the open field. Our findings revealed that ischemic animals spent significantly more time and made more entries in the open arm of the EPM as compared to sham animals, two behaviors indicative of decreased anxiety level. This anxiolytic effect appeared restricted to exploratory models of anxiety, as no differences in punished licking rate were observed between groups in the Vogel/conflict test. In the open field, behavioral monitoring revealed transient ischemia-induced hyperactivity, limited to the initial 15 min of a 30 min testing period. Increased activity in ischemic animals was primarily characterized by increased exploration and sniffing behavior with no significant alterations in rearing and grooming frequencies. Finally, using feeding behavior, our findings revealed a comparable rate of habituation to a novel environment in ischemic and sham rats. Taken together, these results suggest that ischemia-induced hyperactivity may involve a disinhibition to explore unfamiliar and/or mildly anxiogenic environments. However, the basis of such hyperactivity and the presence of habituation deficit following ischemia require further study and/or validation. PMID- 15811390 TI - Re-feeding and the restoration of odor attractivity, odor preference, and sexual receptivity in food-deprived female meadow voles. AB - Food-deprived meadow voles were used to test predictions of two hypotheses associated with the recovery of sexual behaviors following re-feeding. Specifically, we tested between the body weight set point and metabolic fuels hypotheses. To do so, we determined whether the body weight of previously food deprived female voles had to return to pre-food deprivation levels before they would recover their sexual behaviors. The body weight set point hypothesis predicts that food-deprived females that were re-fed will recover their sexual behavior after they return to their original body weight. In contrast, the metabolic fuels hypothesis predicts that food-deprived females that were re-fed will recover their sexual behavior before they return to their original body weight. That is, when the females are in positive energy balance. To distinguish between these two hypotheses, female voles were food deprived for 24 h, which is sufficient to inhibit all three components of sexual behavior. The food-deprived females were then supplied ad libitum food for 0 h, 24 h, 48 h, 72 h, or 96 h and weighed. Females were then tested for their sexual behaviors (odor attractivity, odor preference or proceptivity, sexual receptivity). Re-feeding for 48 h was sufficient to restore odor attractivity, 72 h was sufficient to restore odor preferences for opposite sex conspecific odors, and 96 h was sufficient to restore sexual receptivity to those similar to that of females that were not food deprived. The time-points that the behaviors were recovered were prior to voles recovering their initial body weight. Thus, the data support the metabolic fuels hypothesis. PMID- 15811391 TI - Brain and body temperature homeostasis during sodium pentobarbital anesthesia with and without body warming in rats. AB - High-speed, multi-site thermorecording offers the ability to follow the dynamics of heat production and flow in an organism. This approach was used to study brain body temperature homeostasis during the development of general anesthesia induced by sodium pentobarbital (50 mg/kg, ip) in rats. Animals were chronically implanted with thermocouple probes in two brain areas, the abdominal cavity, and subcutaneously, and temperatures were measured during anesthesia both with and without (control) body warming. In control conditions, temperature in all sites rapidly and strongly decreased (from 36-37 degrees C to 32-33 degrees C, or 3.5 4.5 degrees C below baselines). Relative to body core, brain hypothermia was greater (by 0.3-0.4 degrees C) and skin hypothermia was less (by approximately 0.7 degrees C). If the body was kept warm with a heating pad, brain hypothermia was three-fold weaker ( approximately 1.2 degrees C), but the brain-body difference was significantly augmented (-0.6 degrees C). These results suggest that pentobarbital-induced inhibition of brain metabolic activity is a major factor behind brain hypothermia and global body hypothermia during general anesthesia. These data also indicate that body warming is unable to fully compensate for anesthesia-induced brain hypothermia and enhances the negative brain-body temperature differentials typical of anesthesia. Since temperature strongly affects various underlying parameters of neuronal activity, these findings are important for electrophysiological studies performed in anesthetized animal preparations. PMID- 15811392 TI - Effects of chronic mild stress on sexual behavior, locomotor activity and consumption of sucrose and saccharine solutions. AB - Many symptoms of human depressive disorders are also observed in animals after exposure to unpredictable stressors. The chronic mild stress (CMS) paradigm was developed in order to better model the human situation by using chronic mild stressors over a longer period. It is claimed that the model induces anhedonia in the animals, a core symptom of depression in humans. Despite the fact that the CMS model has a high degree of face validity, there are a number of laboratories in which the establishment of the model is less reliably observed. We have examined behavior (sexual activity and open field activity) together with hedonic measures (sucrose and saccharine intake) after exposure to CMS. CMS decreased male sexual activity (e.g. reduced capability to ejaculate) and increased activity in an open field test. The hedonic measures showed diverging results after CMS in our laboratory. Sucrose consumption was reduced, while saccharine consumption did not show a comparable change. It is concluded that CMS induces comparable alterations to some depression-like symptoms in humans. Saccharine consumption is not a reliable indicator of the hedonic responsiveness to CMS. PMID- 15811393 TI - Differential effects of removing the glucose or saccharin components of a glucose saccharin mixture in a successive negative contrast paradigm. AB - When rats experience an unexpected decrease in reward value, e.g., from 32% sucrose to 4% sucrose, consummatory behavior abruptly decreases to a level below control subjects that only experience the lesser reward, a phenomenon known as Successive Negative Contrast (SNC). In food deprived rats experiencing downshifts in sucrose concentration, SNC dissipates in 3-4 days, as consummatory behavior in shifted rats recovers to the level of unshifted controls. In Experiment 1 food deprived rats that were given 5 min daily access to a 2% glucose-0.15% saccharin mixture, and subsequently shifted to 2% glucose alone, displayed a dramatic SNC effect relative to rats that only received 2% glucose. This SNC effect was primarily manifested as a decrease in the number of consummatory bursts initiated. Interestingly, intake failed to recover to control levels during eight daily postshift sessions. However, in Experiment 2 subjects that were shifted from the same glucose-saccharin mixture to 0.15% saccharin alone failed to show SNC rather, intake fell to the level of control animals which only received 0.15% saccharin. The data from Experiment 1, in conjunction with previous studies utilizing non-deprived rats, quinine adulteration, or shifts from sucrose to saccharin, show that reductions in taste value can produce contrast effects, but suggest that a threshold caloric value is necessary for recovery. The data from Experiment 2 may suggest that saccharin and glucose do not contribute equally to the enhanced palatability of the mixture. PMID- 15811394 TI - Effects of sex and estrous cycle on modulation of the acoustic startle response in mice. AB - Potential sex differences in amplitude, habituation, prepulse inhibition (PPI) and prepulse facilitation (PPF) of the acoustic startle response (ASR) were investigated using male and female mice from the two different inbred mouse strains C57BL/6J (C57) and C3H. Furthermore, the effects of the estrous cycle were tested. The estrous cycle appeared to have no effect on ASR amplitude, habituation, PPF and PPI, the latter being in contrast to results in rats and humans. While sex had no effect on PPI or PPF, males exhibited higher startle amplitudes than females, irrespective of strain, which we discuss to be due to increased male anxiety. In addition, long-term habituation was stronger in C57 males and short-term habituation was weaker in C3H males with respect to females. These results provide evidence for influence of the reproductive hormones on startle reactivity and startle habituation; we therefore conclude that future studies involving genetic influences on behavior using inbred strains are only complete if both sexes are included. PMID- 15811395 TI - Demand-feeding rhythms and feeding-entrainment of locomotor activity rhythms in tench (Tinca tinca). AB - Tench (Tinca tinca) has been described as a strictly nocturnal species whose locomotor activity rhythms, albeit strongly synchronised by light, have an endogenous nature. Aside from light, a number of other environmental factors, such as mealtime, can act as circadian system synchronisers in fish; however, there is a scarcity of information on tench feeding rhythms. This study describes daily self-feeding rhythms in tench, and analyses the role of feeding time on synchronisation of locomotor activity rhythms. Tench were able to operate string sensor-activated self-feeders, and they displayed a strictly nocturnal behavior, both under indoor and outdoor conditions. Locomotor activity remained strictly nocturnal irrespective of whether tench were fed only during the scotophase (D feeding) or the photophase (L-feeding). However, no statistically significant differences were detected between both groups in terms of food intake or growth performance. Furthermore, unlike L-feeding, D-feeding elicited a clear anticipatory activity (FAA). When tench were given the possibility of feeding at both times of the day, they showed a clear preference for D-feeding. Finally, in fish exposed to constant darkness (DD), feeding time acted as a true zeitgeber and FAA was observed. When animals were fasted under DD conditions, locomotor activity free-run and 6 out of 12 individuals yielded significant results in the periodogram analysis. Under DD, fish resynchronised when regular food was resumed, with some tench displaying FAA. The obtained results indicated the existence of a feeding-entrainable oscillator (FEO) in tench. PMID- 15811396 TI - Abnormal patterns of maternal behavior in a genetic animal model of depression. AB - The Flinders Sensitive Line (FSL) model is considered a genetic animal model of depression. Among other characteristics, FSL rats express stress-induced anhedonia and an abnormal dopaminergic system. Our hypothesis was that FSL rats would show abnormal maternal behaviors, especially reduced motivation to reach and care for pups and reduced licking and non-nutritive contact, based on their anhedonic characteristics. Mother-infant interactions were assessed by time limited observations in FSL and Sprague-Dawley (SD) controls. In study 1, differences were found in consummatory behaviors: FSL dams compared to SD dams showed less licking and significant decrease in non-nutritive contact from the first to the third postpartum weeks. In addition, shorter duration of nursing postures was seen in FSL compared to SD dams in the first week postpartum, and this difference was significantly increased by the third week postpartum. In study 2, after exposure to acute swim stress, differences emerged in appetitive behaviors: latencies to reach and care for pups were longer in FSL dams compared to controls, suggesting a stress-induced motivational deficit in FSL dams. Possible explanations, especially regarding the FSL dams' reward system are discussed. PMID- 15811397 TI - Arachidonic acid improves aged rats' spatial cognition. AB - To examine the effects of arachidonic acid (AA) on age-related cognitive deficits, F-344 rats were administered with an AA-supplemented powder diet from 79 weeks of age (OA group). For comparison, we also used an age-matched control group of animals (OC group) that were fed with a non AA-supplemented powder diet. When the subjects reached 87 weeks old, they were trained for Morris water maze place and cue tasks. Escape latencies of the OA group on the place task were significantly shorter than those of the OC group in the latter half of training. The probe test showed that OA rats remembered the trained platform position significantly better than OC rats. In the cue task training, the OC group was significantly slower than the OA group at the beginning of training, but their performance soon matched with that of the OA group. Fatty acids in the hippocampi were measured after the behavioral testing. There was no difference in AA composition in hippocampal phospholipids between the OA and OC groups. However, regression analysis conducted on AA composition and place task performance showed a significant correlation between these two parameters. The present study suggests that AA administration to aged animals can alleviate age-related deficits in spatial cognition. PMID- 15811398 TI - High doses of vasopressin delay the onset of extinction and strengthen acquisition of LiCl-induced conditioned taste avoidance. AB - When low doses of vasopressin are given 50 min after pairing sucrose consumption with a high dose of LiCl, extinction of the LiCl-induced conditioned taste avoidance is accelerated. These low doses of vasopressin do not themselves induce conditioned taste avoidance when paired with sucrose consumption. Predicated on previous studies administering two avoidance-inducing agents after sucrose consumption, studies were designed to determine whether high doses of vasopressin capable of inducing conditioned taste avoidance would (1) delay rather than accelerate extinction of a conditioned taste avoidance induced by a high dose of LiCl and (2) strengthen acquisition of a conditioned taste avoidance induced by a low dose of LiCl. The results of three studies showed that doses of 9 and 18 microg/kg of vasopressin induced a conditioned taste avoidance when injected 50 min after sucrose consumption, delayed the onset of extinction when injected 50 min after pairing sucrose consumption with a high dose of LiCl, and strengthened acquisition of a conditioned taste avoidance when injected 50 min after pairing sucrose consumption with a low dose of LiCl. Taken together, these data suggest that the delay in onset of extinction is due to a strengthening of acquisition. It has been suggested that vasopressin is a mnemonic neuropeptide that delays extinction of learned tasks. However, for conditioned taste avoidance, the evidence for the effects of low doses of vasopressin on extinction do not support this hypothesis and the evidence for high doses of vasopressin can be accounted for by the avoidance-inducing properties of vasopressin. PMID- 15811399 TI - Acute effects of light and darkness on sleep in the pigeon (Columba livia). AB - In addition to entraining circadian rhythms, light has acute effects on sleep and wakefulness in mammals. To determine whether light and darkness have similar effects in birds, the only non-mammalian group that displays sleep patterns comparable to mammals, we examined the effects of lighting changes on sleep and wakefulness in the pigeon. We quantified sleep behavior (i.e., bilateral or unilateral eye closure) in pigeons maintained under a 12:12 LD cycle, and immediately following a change from a 12:12 to a 3:3 LD cycle. During both LD cycles, sleep was most prevalent during dark periods. During the 3:3 LD cycle, darkness had the greatest sleep promoting effect during the hours corresponding to the subjective night of the preceding 12:12 LD cycle, whereas light suppressed sleep across circadian phases. As previously suggested, the light-induced decrease in sleep in the subjective night might be partly mediated by the suppression of melatonin by light. Although the sleep promoting effect of darkness was modulated by the circadian rhythm, sleep in darkness occurred during all circadian phases, suggesting that darkness per se may play a direct role in inducing sleep. In addition to the effects of lighting on behavioral state, we observed an overall bias toward more right eye closure under all lighting conditions, possibly reflecting a response to the novel testing environment. PMID- 15811400 TI - Unmasking of alpha1 adrenoceptor induced hypnogenic response from medial preoptic area. AB - Earlier studies had indicated the possibility of the involvement of the alpha(1) adrenergic receptors on the medial preoptic (mPOA) neurons in hypnogenesis. Microinjection of alpha1 agonist methoxamine (0.05 microg) into the mPOA of the rats, maintained at an ambient temperature (Tamb) of 24 degrees C, did not produce any significant change in sleep-wakefulness (S-W), except for an arousal of short duration, which coincided with the steep fall in body temperature (Tb). It was probably not possible to elicit sleep, because of hypothermia simultaneously produced by this drug. To test this hypothesis, experiments were conducted in an environment where the effects of hypothermia could be partially compensated for. When the rats were allowed to select their Tamb, they moved over to a compartment having a higher Tamb of 30 degrees C, on administration of methoxamine into the mPOA. Simultaneously, there was induction of sleep, which lasted for 60 min. Thus, it was possible to demonstrate the hypnogenic action of methoxamine, which was masked by hypothermic action of the drug. The findings indicate that the alpha1 adrenergic receptors in the mPOA are involved in the induction of sleep. PMID- 15811401 TI - Fish macronutrient selection through post-ingestive signals: effect of selective macronutrient deprivation. AB - Recent reports describe teleosts as being able to regulate energy intake by selecting from pure macronutrient sources, although the regulatory mechanisms involved in this selection remain unknown. The aim of the present work was to determine the effect of selective macronutrient deprivation on energy regulation and macronutrient selection, using for this purpose carbohydrate (CH), protein (P), and fat (F) packaged separately into gelatin capsules, a method that prevents the diet chemosensory properties at oropharyngeal level from interfering with macronutrient selection. Twenty-four individually housed sea bass (Dicentrarchus labrax) were subjected to two experiments: (a) two-macronutrient deprivation, and (b) one-macronutrient deprivation. In two-macronutrient deprivation, fish were fed sequentially with P, CH, or F, and in one macronutrient deprivation, they were fed sequentially with two separately packaged macronutrients (P and CH, CH and F, or P and F). There was a rapid reduction of macronutrient intake in two-macronutrient deprivation, reaching 80% inhibition after 4, 5, and 7 days of P, CH, and F intake, respectively. In one macronutrient deprivation, the energy intake was significantly reduced during selective F deprivation, but not with P or CH deprivation. Although the fish were being fed with only two macronutrients, the relative proportions of these macronutrients in each selective deprivation phase were the same as the baseline. These results show that in deprivation studies fish need at least F plus one other macronutrient to regulate their energy intake, and that their macronutrient selection is stable even when one is absent. In summary, fish seem to regulate energy and macronutrient selection through post-ingestive mechanisms probably involving chemosensory detection in the gut, and/or post-absorptive mechanisms. PMID- 15811402 TI - Deafferentation affects short-term but not long-term control of food intake. AB - Deafferentation affects short-term but not long-term control of food intake (PHYSIOL BEHAV XX(X) 000-000, 2005). Rats were treated neonatally with capsaicin (CAP) to investigate the involvement of vagal afferents in food intake control and body weight regulation. In the first set of experiments, rats were offered increasing concentrations of sucrose (10-15-20-40%) in short-term feeding tests of 1 h. At the end, 10% was offered again to see whether CAP rats modified their intake after repeated exposure to different concentrations of sucrose solution. Results demonstrated that CAP animals overconsume persistently compared to vehicle (VEH) controls. This overconsumption is most pronounced and variable at 10% trials. Hypertonic 40% sucrose solution resulted in a small but significant drop in intake in CAP rats. Overall, if the concentration of sucrose solution is more than 10%, sucrose ingestion of CAP and VEH rats does not depend on the concentration of sucrose solution and remains relatively constant during all trials. In another experiment, rats were exposed to a high-fat condensed milk suspension (CMS) for 5 days. CAP rats initially overconsumed from this CMS compared to VEH. This was accompanied by a decreased intake in chow. However, over the 5 day period CAP animals adjusted their CMS and chow intake to control levels. During both experiments there were no differences in body weight gain between CAP and VEH. Together, these results suggest that capsaicin-sensitive vagal C-fibers are involved in the control of volume ingestion and short-term food intake control but are not required for long-term control of energy intake. PMID- 15811403 TI - Fluidized zero valent iron bed reactor for nitrate removal. AB - A fluidized zero valent iron (ZVI) reactor is examined for nitrate reduction. Using the system, the pH of solution can be maintained at optimal conditions for rapid nitrate reduction. For hydraulic retention times of 15 min, the nitrate reduction efficiency increases with increasing ZVI dosage. At ZVI loadings of 33 gl-1, results indicate that the nitrate removal efficiency increases from less than 13% for systems without pH control to more than 92% for systems operated at pH of 4.0. By maintaining pH at 4.0, we are able to decrease the hydraulic retention time to 3 min and still achieve more than 87% nitrate reduction. The recovery of total nitrogen added as nitrate, ammonium, and nitrite was less than 50% for the system operated at pH4.0, and was close to 100% for a system without pH control. The possibility of nitrate and ammonium adsorption onto iron corrosion products was ruled out by studying the behavior of their adsorption onto freshly hydrous ferric oxide at variable pH. Results indicate the probable formation of nitrogen gas species during reaction in pH4.0. PMID- 15811404 TI - The evaluation of electrical energy per order (E(Eo)) for photooxidative decolorization of four textile dye solutions by the kinetic model. AB - Photooxidative decolorization of four textile dyestuffs, C.I. Acid Orange 7 (AO7), C.I. Acid Orange 8 (AO8), C.I. Acid Orange 52 (AO52) and C.I. Acid Blue 74 (AB74), by UV/H2O2 was investigated in a laboratory scale photoreactor equipped with a 15 W low pressure mercury vapour lamp. The decolorization of the dyes was found to follow pseudo-first-order kinetics, and hence the figure-of-merit electrical energy per order (E(Eo)) is appropriate for estimating the electrical energy efficiency. The E(Eo) values were found to depend on the concentration of H2O2, concentration and basic structure of the dye. This study shows that these textile dyes can be treated easily and effectively with the UV/H2O2 process with E(Eo) values between 0.4 and 5 kW h m-3 order-1, depending on the initial concentrations of dyes and H2O2. The kinetic model, based on the initial rates of degradation, provided good prediction of the E(Eo) values for a variety of conditions. PMID- 15811405 TI - Effect of design parameters in horizontal flow constructed wetland on the behaviour of volatile fatty acids and volatile alkylsulfides. AB - A pilot-scale horizontal flow constructed wetland (HFCW) system planted with common reed (Phragmites sp.) was constructed to study how hydraulic loading rate (HLR), aspect ratio, water depth, and granular medium affect to the fate of several organic matter degradation intermediates namely, acetic acid (HAc), isovaleric acid (Isoval), and dimethylsulfide (DMS). ANOVA statistical analysis performed on the data set of 8 months of operation shows that the HLR and the water depth are two major factors that control the performance of HFCWs for the target analytes. A clear difference in the HFCW effluent concentrations was obtained according to water depth. Effluents of the shallow water depth contained lower DMS (1.05-1.44 microg l-1), HAc (7.91-10.9 mg l-1), and Isoval (0.11-0.15 mg l-1) concentrations than the deeper beds (DMS: 1.68-2.40 microg l-1; HAc: 9.29 14.4 mg l-1, and Isoval: 0.20-0.31 mg l-1). Such differences could be accounted to the different formation and consumption rates of the organic matter degradation intermediates, which is related with the redox potentials (E). Indeed, it could lead to different biochemical reactions of organic matter degradation according with the E value. HLR has a statistically significant influence on the effluent HAc, Isoval, and DMS concentrations. Seasonal variability of effluent HAc concentration shows that it is independent on the HAc loading. While the loading showed no seasonal pattern, the removal efficiency was clearly higher in cold months, which suggests a predominant internal production of HAc in HFCWs in the warm season from the accumulated organic particulate matter. Similar results were also found when Isoval and DMS were considered. PMID- 15811406 TI - Heavy metals removal from electroplating wastewater by aminopropyl-Si MCM-41. AB - The potential of removing nickel and copper from industrial electroplating wastewaters by using mesoporous materials with MCM-41 type structure functionalised with different ratios of aminopropyl groups, namely Na50, Na25 and Na5, were evaluated. The synthesised solids sorbents obtained were characterised by X-ray diffraction, elemental chemical analysis and IR spectroscopy. In preliminary experiments, studies were carried out to determine the optimal experimental conditions for the retention of heavy ions. Effects of concentration, optimal pH, interference with humic substances and other metals were studied for Na5, which showed the best capacity of absorption determined by the corresponding isotherm. This material has a greater selectivity against sodium, indicating that ionic strength does not affect the extraction. Results of an application of this material to remove nickel and copper in synthetic and real industrial wastewater samples from an electrochemical industry area are shown with successful results. The lowest level of nickel and copper were observed when Na5 was used. This observation suggests that reactive aminopropyl-Si MCM-41 and similar materials may be a promising and provide for alternative environmental technologies in the future. PMID- 15811407 TI - Enhanced photocatalytic degradation of VOCs using Ln3+-TiO2 catalysts for indoor air purification. AB - Two types of lanthanide ion-doped titanium dioxide (Ln3+-TiO2) catalysts including La3+-TiO2 and Nd3+-TiO2 were prepared by a sol-gel method. The effects of the lanthanide ion doping on the crystal structure, surface area, adsorption properties, pore size distribution, and surface chemical state of the catalysts were investigated by means of XRD, BET, and XPS. As results, the crystal size decreased significantly, while the specific surface area, t-plot total surface area, micropore volume, and the total pore volume increased owing to the lanthanide ion doping. The nitrogen adsorption-desorption isotherms of the catalysts showed that the N2 adsorption ability of the Ln3+-TiO2 catalysts was better than the TiO2 catalyst. Among them, the 0.7% Ln3+-TiO2 catalysts demonstrated the highest adsorption ability. The photocatalytic activity of the catalysts was investigated in the experiments of the photocatalytic degradation of benzene, toluene, ethylbenzene and o-xylene (BTEX) in a gaseous phase. The photocatalytic efficiency of the TiO2 catalysts with the lanthanide ion doping was remarkably enhanced by BTEX removal. The 1.2% Ln3+-TiO2 catalysts achieved the highest photocatalytic activity. The enhanced photodegradation of BTEX is possibly due to the improved adsorption ability and the enhanced electron-hole pairs separation due to the presence of Ti3+ on the surface of Ln3+-TiO2 catalysts and the electron transfer between the conduction band/defect level and lanthanide crystal field state. PMID- 15811408 TI - Sequential extraction of heavy metals during composting of sewage sludge. AB - The major limitation of soil application of sewage sludge compost is the total heavy metal contents and their bioavailability to the soil-plant system. This study was conducted to determine the heavy metal speciation and the influence of changing the physico-chemical properties of the medium in the course of composting on the concentrations, bioavailability or chemical forms of Cu, Zn, Pb and Ni in sewage sludge. Principal physical and chemical properties and FTIR spectroscopical characterization of sludge compost during treatment show the stability and maturity of end product. The total metal contents in the final compost were much lower than the limit values of composts to be used as good soil fertilizer. Furthermore, it was observed by using a sequential extraction procedure in sludge compost at different steps of treatment, that a large proportion of the heavy metals were associated to the residual fraction (70-80%) and more resistant fractions to extraction X-NaOH, X-EDTA, X-HNO3 (12-29%). Less than 2% of metals bound to bioavailable fractions X-(KNO3+H2O). Heavy metal distribution and bioavailability show some changes during composting depending on the metal itself and the physico-chemical properties of the medium. Bioavailable fractions of all elements tend to decrease except Ni-H2O. Zn and mainly Cu present more affinity to organic and carbonate fractions. In contrast, Pb is usually preferentially bound to sulfide forms X-HNO3. Nickel shows a significant decrease of organic form. Significant degrees of correlation were found between heavy metal fractions and changes of some selected variables (e.g. pH, ash, organic matter, humic substance) during the course of composting. Mobile fractions of metals are poorly predictable from the total content. The R2 value was significantly increased by the inclusion of other variables such as the amount of organic matter (OM) and pH. PMID- 15811409 TI - Simultaneous absorption of NO and SO2 into hexamminecobalt(II)/iodide solution. AB - An innovative catalyst system has been developed to simultaneously remove NO and SO2 from combustion flue gas. Such catalyst system may be introduced to the scrubbing solution using ammonia solution to accomplish sequential absorption and catalytic oxidation of both NO and SO2 in the same reactor. When the catalyst system is utilized for removing NO and SO2 from the flue gas, Co(NH3)(6)2+ ions act as the catalyst and I- as the co-catalyst. Dissolved oxygen, in equilibrium with the residual oxygen in the flue gas, is the oxidant. The overall removal process is further enhanced by UV irradiation at 365 nm. More than 95% of NO is removed at a feed concentration of 250-900 ppm, and nearly 100% of SO2 is removed at a feed concentration of 800-2500 ppm. The sulfur dioxide co-existing in the flue gas is beneficial to NO absorption into hexamminecobalt(II)/iodide solution. NO and SO2 can be converted to ammonium sulfate and ammonium nitrate that can be used as fertilizer materials. The process described here demonstrates the feasibility of removing SO2 and NO simultaneously only by retrofitting the existing wet ammonia flue-gas-desulfurization (FGD) scrubbers. PMID- 15811410 TI - Landfill leachate management in Istanbul: applications and alternatives. AB - Treatment alternatives for Istanbul, Komurcuoda Landfill (KL) leachate that is currently transported to the nearest central wastewater treatment plant were comprehensively investigated with laboratory scale experiments. As flow rate of leachate increases parallel to increment in landfilled solid waste, an individual treatment will be needed to reduce the transportation cost and pollution load on central treatment. However, if the leachate is separately treated and discharged to a brook, in that case more stringent discharge standards will be valid and therefore advanced processes in addition to conventional ones should be included. In laboratory scale experiments, the young landfill leachate having BOD5/COD ratio above 0.6 was successfully treated with efficiencies above 90% in upflow anaerobic reactors if pH is kept below free ammonia inhibition level. Subsequently, nitrification of anaerobically treated leachate was performed with rates of about 8.5 mg NH4+-Ng-1 VSS h-1 and efficiencies above 99% were provided with automated pH regulation by using sodium bicarbonate. Furthermore, denitrification rates as high as 8.1 mg NOx-N g-1VSS h-1 was obtained when carbon source was externally supplied. In addition to nitrification and denitrification, air stripping and struvite precipitation were also applied to remove ammonia in leachate and in average 94% and 98% efficiencies were achieved, respectively. Finally, in average 85% of biologically inert COD was successfully removed by using either ozone or Fenton's oxidation. PMID- 15811411 TI - Biosorption of water-soluble dyes on magnetically modified Saccharomyces cerevisiae subsp. uvarum cells. AB - Brewer's yeast (bottom yeast, Saccharomyces cerevisiae subsp. uvarum) cells were magnetically modified using water based magnetic fluid stabilized with perchloric acid. Magnetically modified yeast cells efficiently adsorbed various water soluble dyes. The dyes adsorption can be described by the Langmuir adsorption model. The maximum adsorption capacity of the magnetic cells differed substantially for individual dyes; the highest value was found for aniline blue (approx. 220 mg per g of dried magnetic adsorbent). PMID- 15811412 TI - Effects of feed solutions on refuse hydrolysis and landfill leachate characteristics. AB - Tap water, aerobically pre-treated leachate, and anaerobically pre-treated leachate, were each fed into the top of a series of three simulated landfills columns, filled with municipal solid waste collected in Shanghai, China. Changes in leachate, including pH, total organic carbon (TOC), and volatile fatty acids (VFAs), and the produced biogas were monitored over time. The tap-water-fed columns had a low hydrolysis rate that yielded an acidic environment (pH 4.8-5.4) in the leachate that inhibited methanogenesis reaction in the refuse. When aerobically pre-treated leachate was fed into the columns, the hydrolysis rate of total organic carbon fluctuated between 200 and 400 mg d-1 and methanogenesis in the refuse column was only partly activated. The hydrolysis rate of refuse fed with anaerobically pre-treated leachate was the highest among the three solutions. The high alkaline levels of the anaerobically pre-treated leachate and its methanogenic bacteria led to an early activation of methanogenesis in the refuse columns. The VFAs contributed approximately 40-60% of TOC in tap-water-fed columns, 60-80% of TOC in the columns fed aerobically pre-treated leachate, and up to 70-90% of TOC in columns fed with anaerobically pre-treated leachate. The feed solution had considerably affected leachate characteristics, and then the build-up of the methanogenesis in the refuse column and the composition of fermentation products in the leachate. The success of a bioreactor landfill depends on whether the recycled leachate could yield a favorable methanogenic environment in the top refuse layer, or whether an appropriate pre-treatment is adopted to modify the leachate characteristics. PMID- 15811413 TI - Evaluation of bioremediation effectiveness on crude oil-contaminated sand. AB - A treatability study was conducted using sea sand spiked with 3% or 6% (w/w) of Arabian light crude oil to determine the most effective bioremediation strategies for different levels of contamination. The sea sand used in the study was composed of gravel (0.1%), sand (89.0%), and silt and clay (10.9%). The water content of the sea sand was adjusted to 12.6% (w/w) for the study. Different combinations of the following treatments were applied to the sand in biometer flasks: the concentration of oil (3% or 6%), the concentration of a mixture of three oil-degrading microorganisms (Corynebacterium sp. IC-10, Sphingomonas sp. KH3-2 and Yarrowia sp. 180, 1x10(6) or 1x10(8) cells g-1 sand), the concentration of the surfactant Tween 80 (1 or 10 times the critical micelle concentration), and the addition of SRIF in a C:N:P ratio of 100:10:3. Three biometer flasks per combination of experimental conditions were incubated, and the performance of each treatment was examined by monitoring CO2 evolution, microbial activity, and oil degradation rate. The results suggest that the addition of inorganic nutrients accelerated the rate of CO2 evolution by a factor of 10. The application of oil-degrading microorganisms in a concentration greater than that of the indigenous population clearly increased biodegradation efficiency. The application of surfactant slightly enhanced the oil degradation rate in the contaminated sand treated with the higher concentration of oil-degrading microorganisms. The initial CO2 evolution rate was shown to efficiently evaluate the treatability test by providing significant data within a short period, which is critical for the rapid determination of the appropriate bioremediation approach. The measurements of microbial activity and crude oil degradation also confirmed the validity of the CO2 evolution rate as an appropriate criterion. PMID- 15811414 TI - The separation of catalyst after photocatalytic reactions conducted in the presence of TiO2/FeCl3/UV. AB - One of the problems connected with wastewater treatment by the photocatalytic method in the presence of TiO2 suspension is necessity of the later catalyst separation. The method proposed by us for this purpose, consists in the usage of coagulation of TiO2 suspension with the aid of FeCl3, particularly in the cases when addition of the salt is used in order to intensification of the photocatalytic process. The effects of the TiO2 separation were studied from the mixtures, after photocatalytic Acid Orange 7 degradation, with dosage of FeCl3 at different stages of the process. The coagulation was carried out at different pH values as well as different FeCl3 concentrations. It was stated that nearly 100% of separation and simultaneously, nearly 100% of decolouration of the examined Acid Orange 7 solutions after their illumination in the presence of TiO2/FeCl3 and coagulation which was produced only by increasing of pH of the obtained mixtures, were nearly possible. PMID- 15811415 TI - Experimental study on the removal of PAHs using in-duct activated carbon injection. AB - This paper presents the incineration tests of municipal solid waste (MSW) in a fluidized bed and the adsorption of activated carbon (AC) on polycyclic aromatic hydrocarbons (PAHs). An extraction and high performance liquid chromatography (HPLC) technique was used to analyze the concentrations of the 16 US EPA specified PAHs contained in raw MSW, flue gas, fly ash, and bottom ash. The aim of this work was to decide the influence of AC on the distribution of PAHs during the incineration of MSW. Experimental researches show that there were a few PAHs in MSW and bottom ash. With the increase of AC feeding rate, the concentrations of three- to six-ring PAHs in fly ash increased, and the concentration of two ring PAH decreased. The total-PAHs in flue gas were dominated by three-, and four ring PAHs, but a few two-, five-ring PAHs and no six-ring PAHs were found. PAHs could be removed effectively from flue gas by using in-duct AC injection and the removal efficiencies of PAHs were about 76-91%. In addition, the total toxic equivalent (TEQ) concentrations of PAH in raw MSW, bottom ash, fly ash, and flue gas were 1.24 mg TEQ kg-1, 0.25 mg TEQ kg-1, 6.89-9.67 mg TEQ kg-1, and 0.36-1.50 microg TEQ Nm-3, respectively. PMID- 15811416 TI - Effect of alkalinity on the performance of a simulated landfill bioreactor digesting organic solid wastes. AB - This study investigated the effects of alkalinity on the anaerobic treatment of the organic solid wastes collected from the kitchen of Engineering Faculty in Dokuz Eylul University, Izmir, Turkey and the leachate characteristics treated in three simulated landfill anaerobic bioreactors. All of the reactors were operated with leachate recirculation. One reactor was operated without alkalinity addition. The second reactor was operated by the addition of 3 g l-1 d-1 of NaHCO3 alkalinity to the leachate and the third reactor was operated by the addition of 6 g l-1 d-1 NaHCO3 alkalinity to the leachate. After 65 d of anaerobic incubation, it was observed that the chemical oxygen demand (COD), volatile fatty acids (VFA) concentrations, and biochemical oxygen demand to chemical oxygen demand (BOD5/COD) ratios in the leachate samples produced from the alkalinity added reactors were lower than the control reactor while the pH values were higher than the control reactor. The COD values were measured as 18900, 3800 and 2900 mg l-1 while the VFA concentrations were 6900, 1400 and 1290 mg l-1, respectively, in the leachate samples of the control, and reactors containing 3 g l-1 NaHCO3 and 6 g l-1 NaHCO3 after 65 d of anaerobic incubation. The total nitrogen (TN), total phosphorus (TP) and ammonium nitrogen (NH4-N) concentrations in organic solid waste (OSW) significantly reduced in the reactor containing 6 g l-1 NaHCO3 by d 65. The values of pH were 6.54, 7.19 and 7.31, after 65 d of anaerobic incubation, respectively, in the aforementioned reactors results in neutral environmental conditions in alkalinity added reactors. Methane percentage of the control, reactors containing 3 g l-1 NaHCO3 and 6 g l-1 NaHCO3 were 37%, 64% and 65%, respectively, after 65 d of incubation. BOD5/COD ratios of 0.27 and 0.25 were achieved in the 3 and 6 g l-1 NaHCO3 containing reactors, indicating a better OSW stabilization. Alkalinity addition reduced the waste quantity, the organic content of the solid waste and the biodegradation time. PMID- 15811417 TI - Sorption of basic and acid dyes from aqueous solutions onto oxihumolite. AB - Naturally occurring kind of weathered and oxidised young brown coal called oxihumolite was used for an adsorptive removal of basic (Methylene Blue, Malachite Green) as well as acid (Egacid Orange, Midlon Black) dyes from waters. It was shown that both kinds of dyes can be sorbed onto oxihumolite. The maximum sorption capacities determined from the parameters of Langmuir isotherms ranged from 0.070 mmol g-1 (for Midlon Black) to 0.278 mmol g-1 (for Malachite Green) and did not differ significantly for basic and acid dyes. The dye sorption (except of Midlon Black) increased in the presence of inorganic salt. Non-ionic surfactants, and surfactants bearing the same charge as the dye exhibited only a minor effect on the dye sorption, whereas oppositely charged surfactants enhanced the dye sorption to a certain extent. The pH value of the aqueous phase exhibited rather pronounced effect on the sorption of acid dyes causing a suppression of the sorption with increasing pH. The sorption of basic dyes, on the other hand, remained almost unchanged in the examined pH range. Oxihumolite is recommended for the treatment of acid wastewaters because of its limited stability in alkaline aqueous solutions. PMID- 15811418 TI - Catalytic reduction of N2O over Ag-Pd/Al2O3 bimetallic catalysts. AB - A study of the catalytic conversion of N2O to N2 over a bimetallic Ag-Pd catalyst is described in this article. Several Ag-Pd catalytic systems were prepared supported on Al2O3 with different ratios and their catalytic activity for the direct decomposition of N2O and their reduction with CO was measured. Based on the experimental results, it was observed that Ag-Pd bimetallic catalyst (5-0.5%) was the most active for both nitrous oxide reduction and direct decomposition. This high activity seems to be connected with a synergistic effect between Ag and Pd. PMID- 15811419 TI - Decolorization of dyes and textile wastewater by potassium permanganate. AB - Decolorization of 10 types of dye solutions by potassium permanganate was studied. Effects of reaction conditions on the decolorization efficiency were examined in batch experiments. The pH value had a significant effect on the decolorization efficiency. When pH value <1.5, the decolorization efficiency was very high. When pH value >4.0, the dye solutions were almost not decolorized. Concentration of potassium permanganate and temperature also showed significant effects on the decolorization efficiency. The decolorization rate of dye solutions by potassium permanganate was rapid, and most of dye solutions can be decolorized effectively. The results of total organic carbon indicated that dye solutions were degraded incompletely by potassium permanganate. The results of treatment of textile wastewater by potassium permanganate indicated that the oxidation with potassium permanganate might be used as a pre-treatment process before biological treatment. PMID- 15811420 TI - Nano-crystal glass-ceramics obtained by crystallization of vitrified red mud. AB - Glass has been obtained by melting red mud from Shandong Province in China with different additives. Suitable thermal treatments were employed to convert the obtained glass into nano-crystal glass-ceramics. X-ray diffraction (XRD) patterns showed that the main crystalline phase in both the glass-ceramics is wollastonite (CaSiO3). These crystals are homogeneously dispersed within the parent glass, with an average crystal size of less than 100 nm. The size of nano-crystals varies when different thermal processes were used. Physical and mechanical properties, such as density, thermal expansion coefficient, hardness, and bending strength, of the two glasses have been examined and the corresponding microstructures are discussed. These results demonstrate that both glass-ceramics have potential for a wide range of construction application. PMID- 15811421 TI - Wet air oxidation of nitrobenzene enhanced by phenol. AB - Simultaneous nitrobenzene and phenol wet air oxidation was investigated in a stainless autoclave at temperature range of 180-220 degrees C and 1.0 MPa oxygen partial pressure. Compared with the single oxidation of nitrobenzene under the same conditions, the presence of phenol in the reaction media greatly improved the removal efficiency of nitrobenzene. The effect of temperature on the reaction was studied. Phenol was considered as a type of initiator in the nitrobenzene oxidation. PMID- 15811422 TI - Asymmetry and the origins of ageing. PMID- 15811423 TI - Age- and sex-dependent alterations in protein kinase C (PKC) and extracellular regulated kinase 1/2 (ERK1/2) in rat myocardium. AB - Cardiovascular morbidity and mortality increase significantly with advancing age, with proportionally higher rates occurring in aged women when compared to aged men. The signaling alterations responsible for age-related reductions in ischemic stress reserves, particularly in aged women, are poorly understood. Accordingly, we sought to determine whether alterations in the cellular location and formation of specific protein kinase C (PKC)-extracellular regulated 1/2 (ERK1/2) signaling modules (SMS) might provide insight into known age- and sex-related differences in cardiovascular disease outcomes. Cytosolic (Cyto), mitochondrial (Mito) and nuclear (Nuc) fractions were isolated from left ventricles of male (M) and female (F) adult (6 mo), castrated or aged (23 mo) F344 rats by centrifugation. Western blotting was used to assess PKC (alpha, delta, epsilon), p-ERK1/2 and p Bad(Ser112) levels, and immunoprecipitation to assess PKC-ERK1/2 SMS. Cyto PKCalpha levels increased with age (p<0.0001), whereas increases in cyto-PKCalpha ERK1/2 SMS were only observed in aged F (60%; p<0.01). Mito-PKCdelta and Mito PKCdelta-ERK1/2 SMS increased in M and F with age (p<0.0001); however increases in Cyto-PKCdelta were only observed in aged M (80% p<0.0001). It is important to note that Nuc- and Mito-PKCdelta-ERK1/2 SMS were 3.5- and 4.8-fold greater in males versus females, respectively (p<0001). Increases in Mito-PKCepsilon-ERK1/2 SMS (216%) were also specific to aged M (p<0.0001), however, Mito-p-Bad(Ser112) levels were decreased with age in both M and F. Differences in sex hormone status could not fully account for observed age-related differences in PKC. Collectively, our results provide novel evidence for age and sex-related differences in the magnitude and distribution of cardiac PKC-ERK1/2 SMS consistent with previously described pathological and protective phenotypes, respectively. PMID- 15811424 TI - Increased caveolin-1, a cause for the declined adipogenic potential of senescent human mesenchymal stem cells. AB - Mesenchymal stem cell (MSC) has drawn much attention in the aspect of tissue renewal and wound healing because of its multipotency. We initially observed that bone marrow-derived human MSCs (hMSCs) divided poorly and took flat and enlarged morphology after expanded in culture over a certain number of cell passage, which resembled characteristic features of senescent cells, well-studied in human diploid fibroblasts (HDFs). More interestingly, adipogenic differentiation potential of hMSCs sharply declined as they approached the end of their proliferative life span. In this study, altered hMSCs were verified to be senescent by their senescence-associated beta-galactosidase (SA-beta-gal) activity and the increased expression of cell cycle regulating proteins (p16(INK4a), p21(Waf1) and p53). Similar as in HDFs, basal phosphorylation level of ERK was also significantly increased in senescent hMSCs, implying altered signal paths commonly shared by the senescent cells. Insulin, a major component of adipogenesis inducing medium, did not phosphorylate ERK 1/2 more in senescent hMSCs after its addition whereas it did in young cells. In senescent hMSCs, we also found a significant increase of caveolin-1 expression, previously reported as a cause for the attenuated response to growth factors in senescent HDFs. When we overexpressed caveolin-1 in young hMSC, not only insulin signaling but also adipogenic differentiation was significantly suppressed with down-regulated PPARgamma2. These data indicate that loss of adipogenic differentiation potential in senescent hMSC is mediated by the over-expression of caveolin-1. PMID- 15811425 TI - Alteration of the TGF-beta/SMAD pathway in intrinsically and UV-induced skin aging. AB - In an effort to characterize transforming growth factor (TGF-beta) signaling and to determine its association with the aging and photoaging processes, we directly compared the expressions of TGF-beta/SMAD in intrinsically aged and photoaged human skin in vivo. By using an RNase protection assay and by immunohistochemistry, we found that the expression levels of TbetaRII mRNA and protein in the epidermis of the forearm (sun-exposed) of the elderly were significantly lower than that of the upper-inner arm (sun-protected) skin of the same individual. In the epidermis, the expressions of Smad7 mRNA in both the intrinsically aged and photoaged skin of the elderly were higher than in the sun protected skin of the young, and this was elevated in the photoaged epidermis. Decreased pSmad2 immunoreactivity was observed in the epidermis of photoaged forearm skin versus matched intrinsically aged skin. This decrease was also found in the epidermis of upper-inner arm skin of the elderly versus the young. These results suggest that the UV-induced down-regulation of TbetaRII and the concerted over-expression of Smad7 may trigger the inhibition of the TGF-beta-induced phosphorylation of Smad2. PMID- 15811426 TI - Acute stress response in calorie-restricted rats to lipopolysaccharide-induced inflammation. AB - Calorie restriction (CR) reduces morbidity and mortality in a wide range of organisms, possibly through the stress response machinery. We analyzed the acute phase response of CR rats to lipopolysaccharide (LPS)-induced inflammatory challenge. Six-month-old male F344 rats, fed ad libitum (AL) or a 30% calorie restricted diet from 6 weeks of age, received an intravenous LPS injection and were then sacrificed between 0 and 8 h. CR attenuated liver injury without reduction in the plasma concentrations of proinflammatory cytokines or nitric oxide (NO). Western blotting analysis of liver tissue demonstrated that CR did not affect the degradation of cytoplasmic I-kappaB and subsequent nuclear translocation of NF-kappaB, a key transcription factor after inflammatory challenge. We also analyzed the liver gene expression profiles at 0, 1 and 4 h with DNA arrays and cluster analysis. Compared with the AL group, CR upregulated the expression of several genes for inflammatory mediators or their related molecules at 0 h, but not at 1 or 4 h. CR downregulated genes for energy or xenobiotic metabolism and stress response proteins at 0 h. At 1 h, the relatively downregulated genes by CR were those for proteases and the ubiquitin-proteasome pathway. The present results suggest that CR attenuates liver injury without suppression of the proinflammatory response, and that the protective effect emerges from constitutively, rather than inductively, expressed gene products. PMID- 15811427 TI - Dinstinct ROS and biochemical profiles in cells undergoing DNA damage-induced senescence and apoptosis. AB - Cellular senescence and apoptosis are both caused by DNA damage stresses, and their severity appears to decide between the two cellular outcomes. In recent studies, it is suggested that these two states may be closely linked and be switched by certain molecular determinants such as p21WAF1 and caspase (Abdelhadi, 2003). However, it is unknown how the pathways to senescence and apoptosis are determined. In addition, although DNA damage stresses frequently accompany cellular accumulation of reactive oxygen species (ROS), how ROS are involved in the decision between the two pathways is unknown. In the present study, MCF-7 cells were induced to senescence or apoptosis by the treatment of varying doses of adriamycin. And, through a series of time course studies, ROS generation profiles and changes in the status of the proteins involved in growth regulation and apoptosis were determined. Significant levels of ROS were produced in senescing cells but not in apoptotic cells. Therefore, senescence is associated with ROS accumulation, but apoptosis is caused independently of ROS. In addition, cells in these two states exhibited quite distinct time course profiles of the proteins, p53, p21WAF1, and E2F1. PMID- 15811428 TI - Inactivation of p38 kinase delays the onset of senescence in rabbit articular chondrocytes. AB - Replicative senescence limits cellular proliferation in vivo and in vitro. Recently, other groups and we reported that p38 kinase plays a key role on the onset of senescence. In this study, we demonstrated that replicative senescence can be delayed in rabbit chondrocytes in vitro by that p38 kinase inactivation. We found that the activity of p38 kinase is elevated in senescent chondrocytes as compared to pre-senescent counterparts. To examine the role of p38 kinase on the onset of senescence, we inactivated the kinase pharmacologically or genetically using either a chemical inhibitor, SB203580, or dominant negative mutant forms of MKK6 and p38 (MKK6A and p38dn, respectively). We show that the inactivation of p38 kinase leads to the stimulation of proliferation, the extension of life span, and a delay in the onset of senescence, thus implying that p38 kinase limits the life span of rabbit articular chondrocytes in vitro. PMID- 15811429 TI - Chronological and physiological ageing in a polar and a temperate mud clam. AB - We investigated chronological and physiological ageing of two mud clams with regard to the "rate of living theory" (Pearl, 1928) and the "free radical theory of ageing" (Harman, 1956). The Antarctic Laternula elliptica (Pholadomyoida) and the temperate Mya arenaria (Myoida) represent the same ecotype (benthic infaunal filter feeders), but differ in maximum life span, 36 and 13 years, respectively. L. elliptica has a two-fold lower standard metabolic rate than M. arenaria, but its life long energy turnover at maximal age is three times higher. When comparing the two species within the lifetime window of M. arenaria, antioxidant capacities (glutathione, catalase) are higher and tissue oxidation (ratio of oxidised to reduced glutathione, lipofuscin accumulation) is lower in the polar L. elliptica than in the temperate mud clam. Tissue redox state in L. elliptica remained stable throughout all ages, whereas it increased dramatically in aged M. arenaria. Our results indicate that metabolic rates and maintenance of tissue redox state are major factors determining maximum lifespan in the investigated mud clams. PMID- 15811430 TI - Mitochondrial ageing of a polar and a temperate mud clam. AB - We investigated mitochondrial ageing in a temperate (Mya arenaria) and an Antarctic (Laternula elliptica) mud clam, with similar lifestyle (benthic filter feeders) but different maximum life spans (MLSP), 13 and 36 years, respectively. The short-lived temperate M. arenaria showed a more pronounced decrease in mitochondrial function (respiration, respiratory control ratio, proton leak, membrane potential) with age than the long-lived Antarctic L. elliptica. H2O2 generation rates at habitat temperature were far higher in the short-lived M. arenaria compared to L. elliptica. Reactive oxygen species (ROS) production as proportion of the mitochondrial oxygen consumption rate (%H2O2/O2) increased significantly with age in M. arenaria, whereas in L. elliptica the proportion remained unchanged. Lower rates of mitochondrial H2O2 generation were presumably due to mild uncoupling as L. elliptica mitochondria showed higher proton leak compared to M. arenaria mitochondria. The results are discussed in to the light of the "Free Radical-Rate of Living theory", (Pearl, R., 1928. The Rate of Living. Alfred Knopf, New York; Harman, D., 1956. Aging: a theory based on free radical and radiation biology. J. Gerontol. 11, 298-300) and the "Uncoupling to Survive" hypothesis (Brand, M.D., 2000. Uncoupling to survive? The role of mitochondrial inefficiency in ageing. Exp. Gerontol. 35, 811-820). PMID- 15811431 TI - Taking musculoskeletal aging out of the bench: do we finally understand frailty? PMID- 15811432 TI - Gene variants for osteoporosis and their pleiotropic effects in aging. AB - The prevalence of osteoporosis is raising worldwide as improving conditions of living and treatment of other common diseases continuously increases life expectancy. Thus, osteoporosis affects most women above 80 years of age and, at the age of 50, the lifetime risk of suffering an osteoporosis-related fracture approaches 50% in women and 20% in men. Numerous genetic, hormonal, nutritional and life-style factors contribute to the acquisition and maintenance of bone mass. Among them, genetic variations explain as much as 70% of the variance for bone mineral density (BMD) in the population. Dozens of quantitative trait loci (QTLs) for BMD have been identified by genome screening and linkage approaches in humans and mice, and more than 100 candidate gene polymorphisms tested for association with BMD and/or fracture. Sequence variants in the vitamin D receptor (VDR), collagen 1 alpha 1 chain (Col1A1), estrogen receptor alpha (ESR1), interleukin-6 (IL-6) and LDL receptor-related protein 5 (LRP5) genes were all found to be significantly associated with differences in BMD and/or fracture risk in multiple replication studies. Moreover, some genes, such as VDR and IL-6, were shown to interact with non-genetic factors, i.e. calcium intake and estrogens, to modulate BMD. Since these gene variants have also been associated with other complex disorders, including cancer and coronary heart disease, they may represent common genetic susceptibility factors exerting pleiotropic effects during the aging process. PMID- 15811433 TI - Transcriptional regulation of chondrocyte maturation: potential involvement of transcription factors in OA pathogenesis. AB - The principle function of articular cartilage is to provide a low friction load bearing surface that facilitates free movement of joints. Maintenance of this surface depends on the maturational arrest of chondrocytes before terminal hypertrophic differentiation occurs [Exp. Cell Res. 216 (1995) 191; Osteoarthritis Cartilage 7 (1999) 389; J. Cell Biol. 139 (1997) 541; J. Cell Biol. 145 (1999) 783]. In contrast to endochondral ossification which involves a programmed process of chondrocyte maturation culminating in terminal hypertrophy and mineralization [Nat. Genet. 9 (1995) 15], articular chondrocytes (ACs) are constrained from completing the maturational program as evidenced by a lack of type X collagen (colX) and alkaline phosphatase expression [Arthritis Res. 3 (2001) 107; Biochem. J. 362 (2002) 473]. Also, ACs are not responsive to factors that impact the maturational process, including bone morphogenetic protein-2 (BMP 2), a potent stimulator of chondrocyte maturation [J. Orthop. Res. 14 (1996) 937]. Factors that constrain AC maturation are only relieved under unique circumstances such as in osteoarthritis (OA), where proliferation and an increase in the expression of hypertrophic hallmarks indicates that the cells have differentiated into a mature phenotype [Calcif. Tissue Int. 63 (2000) 230]. OA may thus involve the functional loss of mechanisms that arrest articular cartilage differentiation. Responsiveness to various growth or systemic factors translates into activation or repression of specific genes through transcriptional mediators. Understanding the downstream mechanisms involved in this process is of paramount importance. Thus, unraveling the molecular interplay between various factors that regulate chondrocyte maturation during OA occurrence and progression is the main focus of ongoing efforts. PMID- 15811434 TI - Role of endocrine-immune dysregulation in osteoporosis, sarcopenia, frailty and fracture risk. AB - Osteoporosis, a key predictor of hip fractures can be treated using a variety of safe and effective interventions. Nevertheless, optimally effective strategies for the prevention of hip fractures must also incorporate efforts to address a broad range of other potentially reversible factors. Hyperthyroidism, anticonvulsants, caffeine and smoking may decrease bone mass and increase fracture risk at any age. In older individuals it is important to also consider additional risk factors, including long-acting benzodiazepines, poor vision and sarcopenia. The presence of sarcopenia, an age-related decline in muscle bulk and quality enhances the risk of frailty and possibly also hip fracture, particularly if associated with diminished functional mobility, lower quadriceps strength and poor balance or body sway. In this review we examine evidence which indicates the presence of endocrine-immune dysregulation in both osteoporosis and sarcopenia. Post-menopausal declines in serum estrogen and androgen levels contribute to increases in local bone levels of cytoclastic cytokines, followed by increased osteoclastogenesis and bone loss. Similarly, the presence of decreased gonadal hormones and IGF-1, combined with unusually high peripheral levels of cytokines, inflammatory mediators and coagulation markers all enhance the risk of sarcopenia and frailty. We propose that a translational research approach which emphasizes common pathophysiologic mechanisms in osteoporosis and sarcopenia could accelerate the speed of discovery of effective strategies for both frailty and hip fracture prevention. PMID- 15811435 TI - Vitamin D in the aging musculoskeletal system: an authentic strength preserving hormone. AB - Until recently, vitamin D was only considered as one of the calciotrophic hormones without major significance in other metabolic processes in the body. Several recent findings have demonstrated that vitamin D plays also a role as a factor for cell differentiation, function and survival. Two organs, muscle and bone, are significantly affected by the presence, or absence, of vitamin D. In bone, vitamin D stimulates bone turnover while protecting osteoblasts of dying by apoptosis whereas in muscle vitamin D maintains the function of type II fibers preserving muscle strength and preventing falls. Furthermore, two major changes associated to aging: osteoporosis and sarcopenia, have been also linked to the development of frailty in elderly patients. In both cases vitamin D plays an important role since the low levels of this vitamin seen in senior people may be associated to a deficit in bone formation and muscle function. In this review, the interaction between vitamin D and the musculoskeletal components of frailty are considered from the basic mechanisms to the potential therapeutic approach. We expect that these new considerations about the importance of vitamin D in the elderly will stimulate an innovative approach to the problem of falls and fractures which constitutes a significant burden to public health budgets worldwide. PMID- 15811436 TI - Mouse models of senile osteoporosis. AB - Little is known about the pathophysiology of normal human and mouse senescence. On the other hand, the pathology of age-related disorders, such as senile osteoporosis, has been investigated. In vivo studies on the pathology of osteoporosis have been conducted primarily in rodents. Although mouse models of senile osteoporosis display some discrepancies relative to their human counterparts with regard to symptoms and pathology, these experimental models are useful and powerful tools for basic and preclinical studies. Here, we review existing mouse models of senile osteoporosis, including those exhibiting premature aging phenotypes, and discuss their pathogenesis, particularly with regard to age-related changes in stem cells. PMID- 15811437 TI - One step forward in the early detection of acute renal failure. PMID- 15811438 TI - Can DOTS control multidrug-resistant tuberculosis? PMID- 15811439 TI - Ethnicity in sexually transmitted infections and sexual behaviour research. PMID- 15811440 TI - Healing and peace making in the middle east: challenges for doctors. PMID- 15811441 TI - Managing asthma during pregnancy. PMID- 15811442 TI - "What, me worry?" Businesses and AIDS at Davos. PMID- 15811443 TI - Firms fill antiretroviral gap in South Africa. PMID- 15811444 TI - Lester Firkins. PMID- 15811445 TI - Epidemiology. PMID- 15811447 TI - ATAC trial update. PMID- 15811448 TI - ATAC trial update. PMID- 15811449 TI - Mohs' micrographic surgery for basal-cell carcinoma of the face. PMID- 15811450 TI - Mohs' micrographic surgery for basal-cell carcinoma of the face. PMID- 15811453 TI - Management of shoulder dystocia. PMID- 15811454 TI - LRRK2 mutations and Parkinsonism. PMID- 15811455 TI - LRRK2 mutations and Parkinsonism. PMID- 15811456 TI - Neutrophil gelatinase-associated lipocalin (NGAL) as a biomarker for acute renal injury after cardiac surgery. AB - BACKGROUND: The scarcity of early biomarkers for acute renal failure has hindered our ability to launch preventive and therapeutic measures for this disorder in a timely manner. We tested the hypothesis that neutrophil gelatinase-associated lipocalin (NGAL) is an early biomarker for ischaemic renal injury after cardiopulmonary bypass. METHODS: We studied 71 children undergoing cardiopulmonary bypass. Serial urine and blood samples were analysed by western blots and ELISA for NGAL expression. The primary outcome measure was acute renal injury, defined as a 50% increase in serum creatinine from baseline. FINDINGS: 20 children (28%) developed acute renal injury, but diagnosis with serum creatinine was only possible 1-3 days after cardiopulmonary bypass. By contrast, urine concentrations of NGAL rose from a mean of 1.6 microg/L (SE 0.3) at baseline to 147 microg/L (23) 2 h after cardiopulmonary bypass, and the amount in serum increased from a mean of 3.2 microg/L (SE 0.5) at baseline to 61 microg/L (10) 2 h after the procedure. Univariate analysis showed a significant correlation between acute renal injury and the following: urine and serum concentrations of NGAL at 2 h, and cardiopulmonary bypass time. By multivariate analysis, the amount of NGAL in urine at 2 h after cardiopulmonary bypass was the most powerful independent predictor of acute renal injury. For concentration in urine of NGAL at 2 h, the area under the receiver-operating characteristic curve was 0.998, sensitivity was 1.00, and specificity was 0.98 for a cutoff value of 50 microg/L. INTERPRETATION: Concentrations in urine and serum of NGAL represent sensitive, specific, and highly predictive early biomarkers for acute renal injury after cardiac surgery. PMID- 15811457 TI - Does DOTS work in populations with drug-resistant tuberculosis? AB - BACKGROUND: Directly observed therapy (DOTS) is the main strategy for prevention and control of tuberculosis worldwide. However, its effect on tuberculosis transmission in populations with moderate rates of drug-resistant disease is not known. METHODS: This population-based prospective study in southern Mexico between March, 1995, and February, 2000, was based on passive case finding and detection of acid-fast bacilli in sputum samples to diagnose pulmonary tuberculosis. We also used cultures, drug-susceptibility testing, bacterial genotyping, and monitoring of treatment outcomes. FINDINGS: We enrolled 436 patients; the HIV seroprevalence rate was 2%. We used three indicators to monitor continuing tuberculosis transmission: the incidence rate of pulmonary tuberculosis, which decreased by 54.4% between 1995 and 2000, from 42.1 to 19.2 per 10(5) population (p=0.00048); the percentage of clustered pulmonary tuberculosis cases, which decreased by 62.6% from 22% to 8% (p=0.02); and the rate of primary drug resistance, which decreased by 84.0% from 9.4 to 1.5 per 10(5) population (p=0.004). Rates of multidrug-resistant (MDR) tuberculosis also decreased (p<0.0001). The case-fatality ratio was 12% for MDR tuberculosis (five of 41), 7% for strains resistant to at least one drug after exclusion of MDR (four of 55), and 3% for pansusceptible strains (nine of 272). There were 13 treatment failures (11%) in 1995 and one (2%) in 2000 (p=0.012). INTERPRETATION: Even in settings with moderate rates of MDR tuberculosis, DOTS can rapidly reduce the transmission and incidence of both drug-susceptible and drug-resistant tuberculosis. However, further interventions, such as drug-susceptibility testing and standardised or individualised treatment regimens, are needed to reduce mortality rates for MDR tuberculosis. PMID- 15811458 TI - Ethnic variations in sexual behaviour in Great Britain and risk of sexually transmitted infections: a probability survey. AB - BACKGROUND: Ethnic variations in the rate of diagnosed sexually transmitted infections (STIs) have been reported in many developed countries. We used data from the second British National Survey of Sexual Attitudes and Lifestyles (Natsal 2000) to investigate the frequency of high-risk sexual behaviours and adverse sexual health outcomes in five ethnic groups in Great Britain. METHODS: We did a stratified probability sample survey of 11161 men and women aged 16-44 years, resident in Great Britain, using computer-assisted interviews. Additional sampling enabled us to do more detailed analyses for 949 black Caribbean, black African, Indian, and Pakistani respondents. We used logistic regression to assess reporting of STI diagnoses in the past 5 years, after controlling for demographic and behavioural variables. FINDINGS: We noted striking variations in number of sexual partnerships by ethnic group and between men and women. Reported numbers of sexual partnerships in a lifetime were highest in black Caribbean (median 9 [IQR 4-20]) and black African (9 [3-20]) men, and in white (5 [2-9]) and black Caribbean (4 [2-7]) women. Indian and Pakistani men and women reported fewer sexual partnerships, later first intercourse, and substantially lower prevalence of diagnosed STIs than did other groups. We recorded a significant association between ethnic origin and reported STIs in the past 5 years with increased risk in sexually active black Caribbean (OR 2.74 [95% CI 1.22-6.15]) and black African (2.95 [1.45-5.99]) men compared with white men, and black Caribbean (2.41 [1.35 4.28]) women compared with white women. Odds ratios changed little after controlling for age, number of sexual partnerships, homosexual and overseas partnerships, and condom use at last sexual intercourse. INTERPRETATION: Individual sexual behaviour is a key determinant of STI transmission risk, but alone does not explain the varying risk across ethnic groups. Our findings suggest a need for targeted and culturally competent prevention interventions. PMID- 15811460 TI - Autonomic peripheral neuropathy. AB - The autonomic neuropathies are a group of disorders in which the small, lightly myelinated and unmyelinated autonomic nerve fibres are selectively targeted. Autonomic features, which involve the cardiovascular, gastrointestinal, urogenital, sudomotor, and pupillomotor systems, occur in varying combination in these disorders. Diabetes is the most common cause of autonomic neuropathy in more developed countries. Autonomic neuropathies can also occur as a result of amyloid deposition, after acute infection, as part of a paraneoplastic syndrome, and after exposure to neurotoxins including therapeutic drugs. Certain antibodies (eg, anti-Hu and those directed against neuronal nicotinic acetylcholine receptor) are associated with autonomic signs and symptoms. There are several familial autonomic neuropathies with autosomal dominant, autosomal recessive, or X-linked patterns of inheritance. Autonomic dysfunction can occur in association with specific infections. The availability of sensitive and reproducible measures of autonomic function has improved physicians' ability to diagnose these disorders. PMID- 15811459 TI - Re-emergence of early pandemic Staphylococcus aureus as a community-acquired meticillin-resistant clone. AB - During the 1950s, the notorious penicillin-resistant clone of Staphylococcus aureus known as phage type 80/81 emerged and caused serious hospital-acquired and community-acquired infections worldwide. This clone was largely eliminated in the 1960s, concurrent with the widespread use of penicillinase-resistant beta lactams. We investigated whether early 80/81 isolates had the genes for Panton Valentine leucocidin, a toxin associated with virulence in healthy young people. Multilocus sequence analysis suggested that descendants of 80/81 have acquired meticillin resistance, are re-emerging as a community-acquired meticillin resistant S aureus (MRSA) clone, and represent a sister lineage to pandemic hospital-acquired MRSA. PMID- 15811461 TI - Post-mortem MRI as an adjunct to fetal or neonatal autopsy. AB - CONTEXT: Understanding what was wrong with a dead fetus or why a newborn child died can help the parents to grieve, while any findings can be used to inform the parents if there is a risk to future pregnancies and the level of risk. This information is mainly obtained from autopsies, but the number of parents agreeing to autopsy has dropped, which has prompted the search for adjuncts to autopsy. STARTING POINT: The UK's Chief Medical Officer outlined the rationale for exploring alternatives to autopsy in 2000 and 2001. After wide consultation, MRI was deemed to offer the most realistic chance of making a contribution. Results so far are promising, but of insufficient depth and quality to make firm recommendations about providing a service in the future. In 2004, the UK Department of Health therefore tendered for this research to be done in fetal and neonatal (and adult) cases, and this work is due to start in 2005. WHERE NEXT? High-quality anatomical information about the brain and spine of fetuses and neonates can be easily obtained with standard MRI scanners. Most developmental and acquired abnormalities can be seen on post-mortem MRI. More work needs to be done on organs not in the central nervous system, and heart malformations, in particular, might present diagnostic problems. A possible outcome could be whole body MRI plus targeted biopsies of abnormalities taken under image guidance as an adjunct to formal autopsy. PMID- 15811462 TI - Promoting Arab and Israeli cooperation: peacebuilding through health initiatives. AB - This article describes a positive experience in building Arab and Israeli cooperation through health initiatives. Over the past 10 years Israeli, Jordanian, and Palestinian health professionals have worked together through the Canada International Scientific Exchange Program (CISEPO). In the initial project, nearly 17,000 Arab and Israeli newborn babies were tested for early detection of hearing loss, an important health issue for the region. The network has grown to address additional needs, including mother-child health, nutrition, infectious diseases, and youth health. Our guiding model emphasises two goals: project-specific outcomes in health improvement, and broader effects on cross border cooperation. Lessons learned from this experience and the model provide direction for ways that health professionals can contribute to peacebuilding. PMID- 15811463 TI - Treatment of sick children during low-intensity conflict. PMID- 15811464 TI - Nutcracker. PMID- 15811465 TI - Structural investigations of the neutral polysaccharide of Linum usitatissimum L. seeds mucilage. AB - The heterogeneity of the purified water-soluble neutral fraction coming from the mucilage extract of the yellow flaxseed was investigated. After fractionation by size-exclusion chromatography, the analyse of the neutral monosaccharides composition showed a mixture of three major families of polymers. They were all identified as arabinoxylans with a constant A/X ratio of 0.24, but varying in their galactose and fucose residues in the side chains. Furthermore, the molecular weight (M(w)) analysis acquired by multi-angle laser light scattered, revealed the association of two high M(w) polymers [5.7x10(6) (11.1%) and 9.3x10(5) (42.4%) g mol(-1)] with a smaller one [3.2x10(5) g mol(-1) (45.3%)]. PMID- 15811466 TI - Bacterial synthesis of polyhydroxyalkanoates containing aromatic and aliphatic monomers by Pseudomonas putida CA-3. AB - Pseudomonas putida CA-3 has the ability to accumulate to high levels unique polyhydroxyalkanoate (PHA) heteropolymers composed of aromatic and aliphatic monomers. The majority of monomers are aromatic making up 98% of the polymer. (R) 3-hydroxyphenylvalerate and (R)-3-hydroxyphenylhexanoate are the most abundant monomers found in polymers accumulated from phenylalkanoic acids with an uneven and even number of carbons on the acyl side chain respectively. PHAs accumulated from phenylvaleric and phenylhexanoic acid were partially crystalline while all other PHAs were amorphous. Significant differences in the yield and PHA content of the cells occurred when different phenylalkanoic acids were supplied as growth substrates. Increasing the initial concentration of the growth substrate increased both the PHA content of the cells and the overall yield (g PHA/g carbon supplied) of PHA accumulated by P. putida CA-3 cells. The highest PHA content (% cell dry wt.) from an aromatic carbon source was 59% when 15mM phenylvaleric acid was supplied as the sole source of carbon and energy. This corresponded to a maximum PHA yield of 0.42 g PHA/g carbon supplied. In and attempt to increase the level of PHA accumulated from related growth substrates acrylic acid was added to the growth medium. However, the addition of various concentrations of acrylic acid to the growth medium had either no effect or decreased the PHA content of the cell accumulated from phenylalkanoic acids by P. putida CA-3. PMID- 15811467 TI - On the properties of agar gel containing ionic and non-ionic surfactants. AB - Rheological and thermal properties of agar sol and gel in presence of various cationic, anionic and non-ionic surfactants are reported. The agar used was from the red seaweed Gelidiella acerosa. The gel strength, viscosity, rigidity (G'), gelling temperature and melting temperature were observed to decrease in presence of non-ionic surfactants whereas these were enhanced in presence of ionic surfactants. TGA studies showed that 1.5% agar gels containing non-ionic surfactants lose water at a lower temperature than the control agar gel whereas gels containing ionic surfactants hold on to water more tenaciously. DSC studies, on the other hand, show that the gel to sol transition occurs at lower temperatures in presence of non-ionic surfactants and at higher temperature in presence of ionic surfactants when compared with the control gel. The non-ionic surfactants, Triton X-100 and Brij 35, enabled relatively concentrated agar extractive to be filtered readily, as a result of which water usage in the process could be reduced by 50%. The surfactant was subsequently removed through freeze-thaw operations to restore the gelling capacity of the agar. The finding that 0.3-0.4% (w/v) sodium lauryl sulfate (SLS) lowers the sol-gel transition temperature from 41 to 36 degrees C without adversely affecting gel strength is another useful outcome of the study that may enable better formulations of bacteriological agar to be prepared. PMID- 15811468 TI - Thermal denaturation of UV-irradiated wet rat tail tendon collagen. AB - The thermal helix-coil transition of UV irradiated collagen in rat tail tendon has been investigated by differential scanning calorimetry. During UVB irradiation the tendons were immersed in water to keep the collagen fibers in a fully hydrated condition at all times. UV irradiation induced changes in collagen which caused both stabilization and destabilization of the triple helix in fibers. The helix-coil transition for non-irradiated collagen occurred near 64 degrees C, for irradiated 1 and 3 h at 66 and 67 degrees C, respectively. After irradiating for longer times (20-66 h) the helix-coil transition peak occurred at much lower temperatures. The peak was very broad and suggested that collagen was reduced by UV to different polypeptides of different molecular weight and different lower thermal stabilities. It was caused by the disruption of a network of hydrogen-bonded water molecules surrounding the collagen macromolecule. PMID- 15811469 TI - Flow behavior of regenerated wool-keratin proteins in different mediums. AB - Keratin is abundantly present in nature and the major component of hair, wool, feather, nail and horns. Dissolution of keratin is often required when non textile applications are demanded. However, the low solubility of keratin in water is the major problem. It becomes unstable and precipitated when stored for a long time. Therefore, it is necessary to find a good solvent that provides high stability and easy processibility. In this research, we used formic acid and dimethylformamide (DMF) to dissolve regenerated keratin protein films. It is shown that formic acid is a good solvent for regenerated keratin proteins for the purpose of storage. Transparent and stable regenerated keratin solution is obtained in formic acid. PMID- 15811470 TI - Solubility and rheological behavior of silk fibroin (Bombyx mori) in N-methyl morpholine N-oxide. AB - As an important direct solvent for cellulose, N-methyl morpholine N-oxide (NMMO) is environmentally friendly, and potentially very economical. Silk fibroin (SF) (Bombyx mori) can also be dissolved directly in NMMO.H2O. However, it is unexpectedly difficult to obtain a silk fibroin solution with a concentration higher than 10wt.% in this way, and extensive degradation of silk fibroin occurs if the dissolution temperature is higher than 110 degrees C. On the other hand, it is found that regenerated silk fibroin (RSF) film is much easier to dissolve in NMMO.H2O than ordinary SF. The RSF in NMMO.H2O can be easily concentrated to a range from 10 to 25wt.%. The structural differences between the degummed silk fiber and the RSF film lead to this different solubility in NMMO.H2O. The rheological behavior of concentrated RSF/NMMO.H2O solutions were also investigated. Regenerated silk fiber was spun from this type of solution, and its strength can reach up to 3.07 cN/dtex. PMID- 15811471 TI - Immobilization of tyrosinase in polysiloxane/polypyrrole copolymer matrices. AB - Immobilization of tyrosinase in conducting copolymer matrices of pyrrole functionalized polydimethylsiloxane/polypyrrole (PDMS/PPy) was achieved by electrochemical polymerization. The polysiloxane/polypyrrole/tyrosinase electrode was constructed by the entrapment of enzyme in conducting matrices during electrochemical copolymerization. Maximum reaction rate (V(max)) and Michaelis Menten constant (K(m)) were investigated for immobilized enzyme. Enzyme electrodes were prepared in two different electrolyte/solvent systems. The effect of supporting electrolytes, p-toluene sulfonic acid and sodium dodecyl sulfate on the enzyme activity and film morphology were determined. Temperature and pH optimization, operational stability and shelf-life of enzyme electrodes were also examined. Phenolic contents of green and black tea were determined by using enzyme electrodes. PMID- 15811472 TI - Packing-based difference of structural features between thermophilic and mesophilic proteins. AB - Twenty pairs of thermophilic and mesophilic proteins were compared in terms of residue packing distribution to obtain structural features related to protein thermostability. Based on residue packing concept, structural features of residues such as residue packing distribution, inner/outer position, secondary structure and water solvation were investigated. The statistical tests revealed that higher frequency in well-packed state of residues, lower frequency in exposed state and higher frequency in well-packed state of inner positioned residues, and higher frequency in well-packed state of 3/10 helix residues could be general structural features thermophilic proteins have. PMID- 15811473 TI - DNA structural transitions induced by divalent metal ions in aqueous solutions. AB - Using methods of IR spectroscopy, light scattering, gel-electrophoresis DNA structural transitions are studied under the action of Cu2+, Zn2+, Mn2+, Ca2+ and Mg2+ ions in aqueous solution. Cu2+, Zn2+, Mn2+ and Ca2+ ions bind both to DNA phosphate groups and bases while Mg2+ ions-only to phosphate groups of DNA. Upon interaction with divalent metal ions studied (except for Mg2+ ions) DNA undergoes structural transition into a compact form. DNA compaction is characterized by a drastic decrease in the volume occupied by DNA molecules with reversible formation of DNA dense particles of well-defined finite size and ordered morphology. The DNA secondary structure in condensed particles corresponds to the B-form family. The mechanism of DNA compaction under Mt2+ ion action is not dominated by electrostatics. The effectiveness of the divalent metal ions studied to induce DNA compaction correlates with the affinity of these ions for DNA nucleic bases: Cu2+>>Zn2+>Mn2+>Ca2+>>Mg2+. Mt2+ ion interaction with DNA bases (or Mt2+ chelation with a base and an oxygen of a phosphate group) may be responsible for DNA compaction. Mt2+ ion interaction with DNA bases can destabilize DNA causing bends and reducing its persistent length that will facilitate DNA compaction. PMID- 15811474 TI - Preparation and characterization of DNA hydrogel bead as selective adsorbent of dioxins. AB - Salmon milt DNA hydrogel beads were synthesized by an inverse suspension polymerization of acrylamide in the continuous phase of cyclohexane. These DNA hydrogel beads in water medium are stable, more than 82% (w/w) of the DNA can be retained in the hydrogel after a sufficient soaking in water. Comparing with normal adsorbents such as activated carbon and alumina, this DNA matrix showed a selective adsorptivity for the dioxin derivatives with planar structure such as dibenzo-p-dioxin (DD), dibenzofuran (DF) and biphenyl (BP). Rinsing with hexane can regenerate the DNA beads after adsorption by the dioxin derivatives, even the adsorption-regeneration process repeated four times, no significant decrease in the dioxin removal capacity was observed. PMID- 15811475 TI - Mg2+ and Ni2+ ion effect on stability and structure of triple poly I.poly A.poly I helix. AB - The effects of Mg2+ and Ni2+ ions on the absorption spectra of IMP, single stranded poly I and three-stranded A2I in solutions with 0.1 M Na+ (pH 7) have been studied. In contrast to Mg2+ ions, the Ni2+ ions affect the absorption spectra of these polynucleotides and IMP. The concentration dependences of the intensity at the extrema in the differential UV spectra suggest that in the region of high Ni2+ concentrations ionic complexes with poly I and A2I are formed, which are characterized by the association constants K'''I = 2000 M(-1) and K'''A2I = 550 M(-1), respectively. The shape of the DUV spectra prompts the conclusion that these complexes are formed due to the inner-sphere interaction of Ni2+ ions with N7 of poly I and A2I presumably due to the outer-sphere Ni2+-O6 interaction. The formation of the complexes leads to destruction of A2I triplexes. The dependences of the melting temperature (T(m)) of A2I on Mg2+ and Ni2+ concentrations have been measured. The thermal stability is observed to increase at the ionic contents up to 0.01 M Mg2+ and only to 2x10(-4) M Ni2+. At higher contents of Ni2+ ions, T(m) lowers and the cooperativity of A2I melting decreases continuously. In all the cases the melting process is the A2I-->A+I+I (3-->1) transition. According to the "ligand" theory, these effects are generated by the energy-advantageous Ni2+ binding to single-stranded poly I (K'''A2I < K'''I) and by the greater number of binding sites which appears during the 3-->1 transition and is entropy-advantageous. PMID- 15811476 TI - Importance of main-chain hydrophobic free energy to the stability of thermophilic proteins. AB - Living organisms are found in the most unexpected places, including deep-sea vents at 100 degrees C and several hundred bars pressure, in hot springs. Needless to say, the proteins found in thermophilic species are much more stable than their mesophilic counterparts. There are no obvious reasons to say that one would be more stable than others. Even examination of the amino acids and comparison of structural features of thermophiles with mesophilies cannot bring satisfactory explanation for the thermal stability of such proteins. In order to bring out the hidden information behind the thermal stabilization of such proteins in terms of energy factors and their combinations, analysis were made on good resolution structures of thermophilic and their mesophilic homologous from 23 different families. From the structural coordinates, free energy contributions due to hydrophobic, electrostatic, hydrogen bonding, disulfide bonding and van der Waals interactions are computed. In this analysis, a vast majority of thermophilic proteins adopt slightly lower free energy contribution in each energy terms than its mesophilic counterparts. The major observation noted from this study is the lower hydrophobic free energy contribution due to carbon atoms and main-chain nitrogen atoms in all the thermophilic proteins. The possible combination of different free energy terms shows majority of the thermophilic proteins have lower free energy strategy than their mesophilic homologous. The derived results show that the hydrophobic free energy due to carbon and nitrogen atoms and such combinations of free energy components play a vital role in the thermostablisation of such proteins. PMID- 15811477 TI - Central nervous system safety of anticholinergic drugs for the treatment of overactive bladder in the elderly. AB - BACKGROUND: Overactive bladder (OAB) is characterized by urgency and increased frequency of micturition, with or without urinary urge incontinence. Anticholinergic agents are important in the treatment of OAB. However, concerns have emerged about their central nervous system (CNS) safety and the associated risk of cognitive impairment. OBJECTIVE: This article describes the CNS adverse effects of anticholinergic drugs used for the treatment of OAB, with particular emphasis on their use in the elderly. Its objective is to help physicians make optimal choices when selecting anticholinergic treatment for OAB. METHODS: : Relevant data from the literature were identified primarily through a MEDLINE search of articles published through December 2003. The search terms included overactive bladder, central nervous system, anticholinergic, and antimuscarinic. This was not intended to be a systematic review, and articles were chosen for inclusion based on their pertinence to the focus on treatment of OAB in the elderly. RESULTS: Several anticholinergic drugs are available for the treatment of OAB, including oxybutymn, tolterodine, trospium chloride, and propiverine (not available in the United States). Among the agents reviewed, penetration of the blood-brain barrier (as predicted by lipophilicity, polarity, and molecular size and structure) is highest for oxybutymn, lower for tolterodine, and lowest for trospium chloride; limited data are available for propiverine. The total anticholinergic drug burden may also be important in determining the potential for CNS adverse effects. The spectrum of anticholinergic CNS adverse effects ranges from drowsiness to hallucinations, severe cognitive impairment, and even coma. The immediate-release (IR) and extended-release (ER) formulations of oxybutynin have been associated with cognitive impairment. In the only published clinical trial that was identified, no significant differences in CNS adverse effects were observed between the IR and ER formulations of tolterodine. There were few clinical data on the use of propiverine in patients with OAB. Trospium chloride has shown favorable CNS tolerability in postmarketing surveillance studies. CONCLUSION: When considering treatment choices for patients with OAB, particularly the elderly, the potential CNS adverse effects of each anticholinergic agent must be weighed against the severity of OAB symptoms. PMID- 15811478 TI - Topiramate monotherapy in epilepsy and migraine prevention. AB - OBJECTIVES: The purposes of this review were to assess the efficacy of topiramate as monotherapy for epilepsy and migraine prevention, describe how it should be used, and give clinical advice on how to manage the practical aspects of dosing, titration, and possible adverse events in these 2 indications. METHODS: We searched the PubMed and BIOSIS databases using the key words topiramate, epilepsy, and migraine from the year 1987 onward, and subsequently focused the search on larger controlled trial studies of topiramate as monotherapy. RESULTS: Studies have evaluated the use of topiramate as monotherapy in the treatment of partial-onset and generalized seizures and in the prevention of migraine. In a randomized study, 75% of epilepsy patients treated with 400 mg/d topiramate remained seizure free at 1 year. Patients in the same study treated with a lower dose of topiramate (50 mg/d) also experienced notable seizure reductions, with 59% of patients free of seizures at 1 year. A comparison trial of topiramate (100 or 200 mg/d), valproate, and carbamazepine found that topiramate was associated with a similar time to first posttreatment seizure as the other 2 agents (P = NS). Trials of topiramate monotherapy in migraine prevention found that 100 mg/d was associated with a > or =50% reduction in monthly migraine frequency in 49% to 54% of patients. The migraine prevention trials typically used a starting dose of 25 mg/d, with weekly increases of 25 mg and an initial monotherapy target dose of 100 mg/d. The most common adverse events associated with topiramate are paresthesia, weight loss, and other centrally mediated symptoms, many of which may be ameliorated by proper titration and dosing and by good communication between physician and patient. CONCLUSIONS: Data from controlled trials suggest that 100 mg/d topiramate as monotherapy is effective in the treatment of partial onset and generalized seizures and in the prevention of migraine. PMID- 15811479 TI - Manidipine versus enalapril monotherapy in patients with hypertension and type 2 diabetes mellitus: a multicenter, randomized, double-blind, 24-week study. AB - BACKGROUND: Blood pressure reduction is associated with a reduced risk for cardiovascular events and death, particularly in patients with both hypertension and type 2 diabetes mellitus. OBJECTIVE: The aim of this study was to compare the antihypertensive efficacy, tolerability, and effect on metabolic risk factors of manidipine, a new dihydropyridine calcium channel antagonist, and enalapril, a widely used angiotensin-converting enzyme inhibitor, in patients with mild to moderate essential hypertension and type 2 diabetes. METHODS: This multicenter, double-blind trial compared manidipine and enalapril in patients with type 2 diabetes and hypertension (diastolic blood pressure [DBP] 90-104 mm Hg, systolic blood pressure [SBP] < or =190 mm Hg). Following a 3-week, single-blind placebo run-in period, eligible patients were randomized to receive either manidipine 10 mg or enalapril 10 mg once daily for 24 weeks. The dose was doubled after 3 weeks in patients who had not responded to treatment (DBP > or =90 mm Hg). The primary efficacy end point was change in DBP from baseline to the end of the study. Secondary outcomes were the responder rate (DBP <90 mm Hg and/or a DBP reduction of > or =10 mm Hg) at the end of the study. Other secondary measures were changes from baseline to the end of the study in heart rate and in the following measures obtained by ambulatory blood pressure monitoring (ABPM): 24-hour, daytime, and nighttime mean DBP and SBP, and the trough:peak ratio. Blood glucose, glycosylated hemoglobin (HbA1c), total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, uric acid, and creatinine were measured at the end of the placebo run-in period and the end of treatment. The study had 80% power to detect a between-treatment difference in mean sitting DBP of >3 mm Hg. RESULTS: One hundred twenty-four patients were enrolled in the study. After the placebo run-in period, 13 patients were excluded from the study: 4 for DBP values outside the specified limits, 7 at their request, and 2 for adverse events. Thus, 111 patients met the eligibility criteria and were randomized to treatment (53 manidipine, 58 enalapril). The population consisted of 61 men and 50 women with a mean (SD) age of 62 (11) years and a body mass index of 28.2 (2.4) kg/m2. Among patients who completed the study, drug doses were doubled in 67.6% (25/37) of patients in the manidipine group and 60.0% (24/40) of patients in the enalapril group (P = NS). Similar reductions in blood pressure were observed in both groups, from a mean (SD) of 164 (12)/97.5 (5) mm Hg at baseline to 141 (12)/84.5 (6) mm Hg at the end of the study in the manidipine group (P < 0.01), and from 159 (12)/98 (4) mm Hg to 139 (12)/86 (8) mm Hg in the enalapril group (P < 0.01). The proportion of responders was 66.7% (32/48) in the manidipine group and 60.0% (30/50) in the enalapril group; the difference between groups was not significant. Twenty-four-hour ABPM revealed significant (P < 0.01) and similar reductions in blood pressure in both groups, with a trough:peak ratio of approximately -50%. Neither drug affected heart rate. Among the statistically significant changes in metabolic parameters, significant reductions in HbA(1c) (from 6.7% [1.4%] to 6.2% [1.1%]) and blood glucose concentrations (from 152 [44] to 143 [44] mg/dL) were observed only in the manidipine group (P < 0.05). The incidence of adverse events was similar between groups. CONCLUSIONS: In the present study, manidipine was as metabolically neutral and as effective as enalapril in reducing blood pressure in hypertensive patients with type 2 diabetes, providing a sustained 24-hour antihypertensive effect. PMID- 15811481 TI - Double-blind, randomized, parallel, placebo-controlled study of ibuprofen effects on thromboxane B2 concentrations in aspirin-treated healthy adult volunteers. AB - BACKGROUND: Patients taking aspirin for cardioprotection may occasionally take over-the-counter (OTC) ibuprofen for pain relief, which might interfere with the antiplatelet effects of aspirin. OBJECTIVE: The present study was undertaken to determine whether ibuprofen, taken according to OTC label directions, would affect inhibition of thromboxane B2 (TXB2), a surrogate for platelet inhibition. METHODS: This was a prospective, multiple-dose,single-center, double-blind, randomized, parallel, placebo-controlled study. Eligible subjects received chewable, immediate-release aspirin 81 mg QD for 8 days, and were then randomized to receive either ibuprofen 400 mg TID or placebo TID, in addition to aspirin, for 10 days. RESULTS: Fifty-one subjects were randomized; 47(24 placebo, 23 ibuprofen) completed the study. No subjects withdrew prematurely. Subjects were predominantly white (49%) or black (38%), and 53% were male. The mean (SD) age was 38.4 (9.8) years and mean (SD) body weight was 173.2 (26.7) pounds. On days 1, 3, 7, and 10 of the study period, mean TXB2 inhibitions were 99.24%, 98.88%, 97.75%, and 98.17% for ibuprofen and 98.82%, 98.93%, 98.75%, and 98.83% for placebo. Although a statistically significant reduction of TXBZ inhibition was seen in the ibuprofen group at days 7 and 10 (P = 0.003 and P = 0.023, respectively), TXBZ inhibition was >90% on all days tested in all subjects. Aspirin, ibuprofen, and placebo were all well tolerated. There were 3 adverse events (1 mild and 2 moderate) during the aspirin run-in period and 8 (2 mild and 6 moderate) during the randomized study period. CONCLUSIONS: No clinically meaningful loss of cardioprotection was found, as reflected by TXB2 inhibition in healthy volunteers who received OTC doses of ibuprofen. When using this regimen of OTC ibuprofen with immediate-release, low-dose aspirin, concerns about the loss of cardioprotective antiplatelet effects of aspirin are not supported by this study. PMID- 15811480 TI - Long-term safety and, tolerability profiles and lipid-modifying efficacy of ezetimibe coadministered with ongoing simvastatin treatment: a multicenter, randomized, double-blind, placebo-controlled, 48-week extension study. AB - BACKGROUND: Ezetimibe (EZE) is a cholesterol-lowering drug that inhibits absorption of dietary and biliary cholesterol across the intestinal wall without affecting absorption of bile acids, fatty acids, fat-soluble vitamins, or triglycerides. It has a complementary mechanism of action to the statins, which inhibit cholesterol synthesis in the liver. Coadministration of EZE and statins provides inhibition of 2 sources of cholesterol, leading to greater reductions in low-density lipoprotein cholesterol (LDL-C) than with either agent alone. OBJECTIVES: This study evaluated the long-term safety and tolerability profiles and lipid-modifying efficacy of treatment with EZE 10 mg/d plus simvastatin (SIMVA) 10, 20, 40, or 80 mg/d for 48 weeks in patients with primary hypercholesterolemia. METHODS: This was an extension of a multicenter, double blind, placebo (PBO)-controlled base study in which hypercholesterolemic patients were randomized to receive EZE 10 mg/d or PBO in addition to their current statin for 8 weeks. Patients who successfully completed the base study could enter the extension study if they were willing to switch from their current statin to an approximately equipotent dose of SIMVA for the 54-week study period. After a 6 week open-label SIMVA run-in phase, patients were rerandomized to receive EZE 10 mg/d or PBO in a 4:1 ratio, respectively, for 48 weeks. At each clinic visit, beginning at week 12, the dose of SIMVA was titrated upward until patients reached their National Cholesterol Education Program Adult Treatment Panel II LDL C goal or the maximum SIMVA dose of 80 mg/d. Safety/tolerability and lipid efficacy parameters were assessed at 12-week intervals. RESULTS: Of 433 patients entering the extension study, 355 were randomized to receive EZE and 78 were randomized to receive PBO. Baseline demographic characteristics and lipid levels were similar between treatment groups. Overall, coadministration of EZE + SIMVA was well tolerated. There were no clinically meaningful differences between the EZE and PBO groups with regard to the incidence of treatment-related adverse events (AEs) (19% vs 17%, respectively), discontinuations due to AEs (7% vs 10%), serious AEs (12% vs 17%), consecutive elevations in liver function tests > or =3 times the upper limit of normal (ULN) (0.3% vs 0%), or elevations in creatine kinase > or =10 times the ULN (both, 0%). As in the base study, LDL-C levels were significantly lower with the addition of EZE to SIMVA compared with the addition of PBO (-24% vs 3%; P < 0.001). CONCLUSION: In these patients with primary hypercholesterolemia, EZE 10 mg/d added to ongoing SIMVA treatment for 48 weeks had a favorable safety and tolerability profile and was more efficacious than SIMVA monotherapy. PMID- 15811482 TI - Vasopressor administration during liver transplant surgery and its effect on endotracheal reintubation rate in the postoperative period: a prospective, randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: End-stage liver disease (ESLD) is associated with a low systemic vascular resistance due to peripheral vasodilatation. This phenomenon is aggravated by general anesthesia (GA) administered during liver transplantation, resulting in precipitous decreases in blood pressure. The excessive amounts (>3 mL/1 mL blood loss) of IV fluid administered to maintain hemodynamic stability during surgery promotes a fluid shift in the lung, which may lead to hypoxia in the immediate postoperative period. This pathophysiologic state may necessitate endotracheal reintubation and mechanical ventilation of the lungs, thus exposing the patient to a risk for morbidities related to laryngoscopy and endotracheal intubation, including deleterious cardiovascular responses to laryngoscopy, endotracheal damage due to laryngoscopic instrumentation, alteration in pulmonary mechanics secondary to controlled mechanical ventilation of the lungs, and delayed recovery associated with the sedation needed to perform these maneuvers. OBJECTIVE: The aim of this study was to determine whether the use of a vasopressor to antagonize the vasodilatory effect of GA would reduce the amount of IV fluids administered during liver transplantation, and whether the subsequent amelioration of fluid shift in the postoperative period would reduce the need for ventilatory support and endotracheal reintubation. METHODS: This prospective, randomized, double-blind, placebo-controlled study was conducted at the University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, New Jersey. Patients aged > or =18 years scheduled to undergo orthotopic liver transplantation for ESLD were enrolled. The effect of use of an adjuvant vasopressor, together with controlled fluid administration (ie, the volume of IV fluid needed to maintain hemodynamic parameters at > or =80% of preoperative levels) (vasopressor group), was compared with that of fluid administration only (placebo group). We determined various postoperative outcome measures, primarily the amount of fluid administered and the need for endotracheal reintubation. RESULTS: Sixty-five patients were enrolled (44 men, 21 women; vasopressor, 33 patients; placebo, 32 patients). Sex distribution showed 19 men and 14 women in the vasopressor group and 25 men and 7 women in the placebo group (both, P < 0.05). The 2 treatment groups were statistically similar with regard to the rest of the baseline demographic and clinical characteristics and duration of surgery. The vasopressor group had a significantly lower prevalence of endotracheal reintubation compared with the placebo group (RR, 1:6; P < 0.05). The other postoperative parameters were statistically similar between the 2 groups. CONCLUSION: In this study of adults undergoing orthotopic liver transplantation for ESLD, use of an adjuvant vasopressor, together with controlled fluid administration, to maintain a stable hemodynamic status during GA reduced the need for endotracheal reintubation and its associated morbidities in the postoperative period compared with placebo. PMID- 15811483 TI - Impact of an adherence clinic on behavioral outcomes and virologic response in treatment of HIV infection: a prospective, randomized, controlled pilot study. AB - OBJECTIVE: The aim of this randomized, controlled pilot study was to examine the impact of a pharmacist operated adherence clinic on adherence to highly active antiretroviral therapy (HAART) and viral suppression in patients with HIV over 28 weeks. METHODS: Consecutive eligible patients initiating HAART at an indigent care clinic were randomized to an adherence clinic or to standard care (information provided by physician or nurse practitioner) for education and monitoring. Group assignment was stratified before randomization according to regimen complexity and potential tolerability. Adherence (electronic monitoring and patient self-report) and viral load (reverse-transcription polymerase chain reaction) were assessed at weeks 4, 16, and 28. RESULTS: Thirty-three randomized patients (adherence clinic, n = 16; standard care, n = 17) comprised the intent to-treat population. The groups were well-matched for demographics and antiretroviral regimen. The median age was 38.0 years in both groups. Most patients were male (85%), had previously used HAART (78%), and had an AIDS diagnosis (79%). Mean (SD) adherence at weeks 4, 16, and 28 was 86% (27%), 77% (28%), and 74% (31%) in the adherence clinic group versus 73% (32%), 56% (39%), and 51% (41%) in the standard care group (week-16 difference, 21% [90% CI, 1% 42%]; week-28 difference, 23% [90% CI, 1%-44%]). Sixty-nine percent of patients in the adherence clinic group took their medication on schedule versus 42% in the standard care group (P = 0.025); mean decline in adherence from weeks 4 to 28 was 12% in the adherence clinic group (P = 0.15) versus 22% in the standard care group (P = 0.002). HIV-1 RNA levels were <400 copies/mL at weeks 4, 16, and 28 in 63%, 100%, and 94% of the adherence clinic group and 29% (P = NS), 71% (P = 0.04), and 65% (P = NS) of the standard care group. CONCLUSIONS: In this preliminary trial, an adherence clinic model improved adherence to HAART and virologic response over 28 weeks in the patients studied. PMID- 15811484 TI - A randomized, open-label, crossover study examining the effect of injection site on bioavailability of exenatide (synthetic exendin-4). AB - BACKGROUND: Exenatide (synthetic exendin-4;AC2993) is a 39-amino acid peptide in the new class of antidiabetic agents known as incretin mimetics. In clinical trials, exenatide exhibited glucoregulatory effects (glucose-dependent stimulation of insulin secretion, suppression of inappropriately elevated glucagon secretion, slowing of gastric emptying) in patients with type 2 diabetes mellitus (DM). OBJECTIVE: The goal of this study was to determine the relative bioavailability of exenatide injected subcutaneously into the abdomen, arm, or thigh. METHODS: Patients with type 2 DM were randomized in an open-label, crossover study to assess relative bioavailability of exenatide (10 microg) injected into the arm and thigh versus injection into the abdomen. Serial plasma exenatide concentrations were measured for 10 hours after injection. A sample size of >24 patients provided approximately 80% power to ensure that 90% CIs were within the 80% to 125% interval for the ratios (geometric least squares [LS] means) of AUC(0-infinity). RESULTS: Twenty-eight patients were randomized into the study (mean age, 56 [8] years; glycosylated hemoglobin, 8.0 [1.7]%; body mass index, 33 [5] kg/m2; all values given as mean [SD]). AUC(0-infinity) values (geometric LS mean SE for SC injections into the abdomen arm and thigh were 63,935 (6608), 59,573 (6157), and 62,148 (6424) pg./mL, respectively. The AUC (geometric LS mean ratio for relative bioavailability) for arm versus abdomen was 0.93 (geometric 90% CI, 0.82-1.05); for thigh versus abdomen it was 0.97 (geometric 90% CI, 0.86-1.10). Consistent with the observed data, intrasubject variability of AUC(0-infinity) was low among the 3 treatments (coefficient of variation, 26%). C(max) values (geometric LS mean [SE]) were 220 (24) pg/mL, abdomen; 218 (23) pg/mL, arm; and 193 (21) pg/mL, thigh. The C(max) (geometric LS mean ratio) for arm versus abdomen was 0.99 (geometric 90% CI, 0.85-1.15), and for thigh versus abdomen it was 0.88 (geometric 90% CI, 0.75-1.02). The most common treatment-emergent adverse events were mild to moderate nausea (36%), headache (25%), vomiting (21%), and dizziness (18%). Three patients received an inadvertent 10-fold overdose and were withdrawn from the study immediately. All experienced severe nausea and vomiting, and 1 patient experienced severe hypoglycemia requiring aid. All recovered without mishap and were excluded from statistical and tolerability results. There were no adverse events related to the injection or the injection site. CONCLUSION: In this study of patients with type 2DM, SC administration of exenatide into the abdomen, arm, or thigh resulted in comparable bioavailability. PMID- 15811485 TI - Topical retinoids in inflammatory acne: a retrospective, investigator-blinded, vehicle-controlled, photographic assessment. AB - BACKGROUND: Despite published data showing the efficacy of topical retinoids in treating inflammatory acne, in clinical practice topical retinoids tend to be used most commonly for noninflammatory acne. OBJECTIVE: The goal of this study was to assess the efficacy of topical retinoids as monotherapy in inflammatory acne. METHODS: This retrospective, investigator-blinded,vehicle-controlled, photographic assessment study was conducted by 5 investigators. The investigators rated pretreatment and posttreatment photographs of patients who had participated in 12- or 15-week, double-blind comparisons of tazarotene 0.1% gel, adapalene 0.1% gel, tretinoin 0.1% microsponge, tretinoin 0.025% gel, and tazarotene 0.1% cream (vehicle). Acne severity was rated on a 7-point scale. A posttreatment increase or decrease of 1 grade was considered clinically meaningful; > or =2 grades was considered an even clearer measure of clinically significant improvement. Investigators also rated global response to treatment on a 7-point scale. A posttreatment increase of > or =2 grades was considered a clinically relevant improvement. RESULTS: Each of the 5 investigators rated photographs of 577 patients ( approximately 52% women, approximately 48% men; mean age, 18-20 years), for a total of 2885 evaluations (in addition to daily evaluations of the 20 control patients). The treatment groups consisted of tazarotene (252 patients, 1260 evaluations), adapalene (178 patients, 890 evaluations), tretinoin microsponge (47 patients, 235 evaluations), tretinoin gel (39 patients, 195 evaluations), and vehicle (61 patients, 305 evaluations). Inflammatory acne was improved with all 4 retinoids compared with vehicle. In 1905 evaluations in which pretreatment acne severity was grade > or =3 (mild to moderate), the incidences of clinically significant improvements in the tazarotene, adapalene, and tretinoin microsponge groups were 24%, 17%, and 21%, respectively (all, P < or = 0.001 vs vehicle [7%]). The difference in prevalence of clinically significant improvement was statistically similar between the tretinoin gel and vehicle groups. The incidences of clinically relevant improvement in global response to tazarotene, adapalene, tretinoin microsponge, and tretinoin gel were 36%, 34%, 31%, and 28%, respectively (P < or = 0.001, < or =0.001, < or =0.001, and < or =0.01, respectively, vs vehicle [17%]). CONCLUSIONS: : The results of this study suggest that topical retinoid monotherapy can achieve clinically significant improvements in inflammatory acne. PMID- 15811486 TI - Equipotent doses of transdermal fentanyl and transdermal buprenorphine in patients with cancer and noncancer pain: results of a retrospective cohort study. AB - BACKGROUND: The equipotency ratio of transdermal (TD) fentanyl to oral morphine has been established as 1:100; for buprenorphine TD, a ratio of 1:75 has been proposed, although this ratio has not been confirmed in clinical studies. Growing evidence from clinical practice, in which much lower doses of buprenorphine are used, suggests that this conversion ratio may be too high. OBJECTIVE: The aim of this study was to compare calculated equipotent oral morphine doses of fentanyl TD with equipotent oral morphine doses of buprenorphine TD prescribed in clinical practice. METHODS: This retrospective study identified patients with cancer and noncancer pain who had received > or =1 prescription for fentanyl TD or buprenorphine TD (the all-patients groups) from the German IMS Disease Analyzer mediplus database, which contains all relevant data concerning drug prescriptions from 400 practices in Germany. Also identified were subgroups of the all-patients groups who had received long-term treatment with fentanyl TD or buprenorphine TD and were considered to have similar pain intensity, as they had previously received similar analgesic medication (the identical-cohort groups). Mean prescribed daily doses for the all-patients and identical-cohort groups were calculated based on the distribution of prescribed patch strengths. Because patients could have applied >1 patch, mean prescribed daily doses were also calculated based on an assumption of double application when appropriate. Equipotent oral morphine doses were estimated using equipotency ratios of 1:100 for fentanyl TD and 1:75 for buprenorphine TD. RESULTS: The all-patients groups consisted of 2198 patients with noncancer pain and 2544 patients with cancer pain; the identical-cohort groups consisted of 380 patients with noncancer pain and 496 patients with cancer pain (529 women, 347 men; mean age, 74 years [range, 25-101 years]). Equipotent doses of oral morphine were significantly lower in patients receiving buprenorphine TD compared with those receiving fentanyl TD (P < 0.001). In cancer patients, the equipotent oral morphine doses of fentanyl TD and buprenorphine TD were 130.9 to 138.9 mg and 85.2 to 88.8 mg, respectively; in noncancer patients, the corresponding values were 117.0 to 118.3 mg and 80.2 to 80.9 mg. Based on these results, an equipotency ratio of 1:110 to 1:115 for buprenorphine TD would appear to be more appropriate than the proposed ratio of 1:75. CONCLUSIONS: The fact that this retrospective analysis conducted in identical cohorts showed lower calculated equipotent oral morphine doses in the buprenorphine TD groups compared with the fentanyl TD groups calls into question the proposed 1:75 ratio for conversion of buprenorphine TD to equipotent oral morphine doses. Based on the findings of the present study, an equipotency ratio of 1:110 to 1:115 may be more appropriate. However, confirmative data from prospective randomized clinical trials are needed. PMID- 15811487 TI - Assessment of the impact of a patient clinical trials handbook among pharmacy students. AB - BACKGROUND: Patient education in the basic concepts of clinical trials is necessary to promote understanding of the informed consent process and enhance patients' decision-making. It has been suggested that patients' knowledge and attitudes are improved by being given general written information about clinical trials. OBJECTIVE: This pilot study was conducted to determine the effect of a patient education handbook on the knowledge, attitudes, and motivations of pharmacy students regarding clinical trials. METHODS: A patient clinical trials handbook was developed at a 7th-grade reading level for the Department of Veterans Affairs Cooperative Studies Program and tested in PharmD students. Students were randomized to the experimental group (received handbook) or the control group (no handbook). They were given 15 to 20 minutes to read the handbook, after which they were asked to respond to a questionnaire adapted from previous studies. The questionnaire included 25 true/false questions testing participants' knowledge of clinical trials, 5 questions on attitudes toward clinical trials scored on a 5-point Likert scale, and 6 questions concerning their motivation toward participation in hypothetical clinical trial scenarios scored on a 5-point Likert scale. The experimental group was also asked to rate the informativeness, helpfulness, and clarity of the handbook on a 5-point Likert scale. RESULTS: There were 40 students in the experimental group and 50 in the control group. Knowledge scores were significantly higher in the experimental group compared with the control group (mean [SD] percentage of correct answers, 88.7% [8.0%] vs 82.6% [9.0%], respectively; P < 0.001). Positive attitudes toward clinical trials were also increased in the experimental group compared with the control group; specifically, participants expressed significantly greater clarity of understanding of clinical trials (mean score, 1.4 [0.5] vs 0.8 [0.6]; P < 0.001) and relief associated with knowing about clinical trials (mean score, 0.8 [0.8] vs 0.4 [0.7]; P = 0.017). There were no between-group differences in students' motivation to participate in the hypothetical clinical trial scenarios. A high proportion of students (95%) found the handbook informative, helpful, and understandable. CONCLUSIONS: The patient clinical trials handbook increased knowledge and positive attitudes regarding clinical trials among pharmacy students participating in this study. PMID- 15811488 TI - Treatment of hypoglycemia using combined glucocorticoid and recombinant human growth hormone in a patient with a metastatic non-islet cell tumor hypoglycemia. AB - BACKGROUND: Non-islet cell tumor hypoglycemia(NICTH) is a rare cause of recurrent hypoglycemia. It has been associated with the tumoral overproduction of high molecular-weight insulin-like growth factor (IGF)-2 ("big IGF-2"). Big IGF-2 suppresses growth hormone (GH) biosynthesis and impairs the storage of IGFs by suppressing the formation of the GH-dependent ternary complexes containing IGF, IGF binding protein 3 (IGFBP-3), and acid-labile subunit (ALS). Thus, big IGF-2 exerts hypoglycemic activity. The only effective treatment of NICTH is surgery. However, in inoperable patients with NICTH, treatment of hypoglycemia may require high doses of glucocorticoid (30-60 mg/d [0.5-1.0 mg/kg x d]) or recombinant human GH (rhGH) (2.6-12.0 mg/d [0.043-0.20 mg/kg x d]). OBJECTIVE: We hypothesized that the association of low doses of glucocorticoid and rhGH could be an effective therapy for hypoglycemia in inoperable patients with NICTH. METHODS: A 3-phase treatment regimen was conducted in an inoperable 67-year-old woman with NICTH. Decreasing dosages of prednisone (from 30 to 10 mg/d [from 0.50 to 0.15 mg/kg x d]), followed by decreasing doses of rhGH (from 2.6 to 1.3 mg/d [from 0.043 to 0.016 mg/kg x d]), and then a combination of the lowest doses of each, were tested. Glucose, insulin, and IGF monitoring were performed at each of the 3 treatment phases. RESULTS: Fasting plasma glucose (FPG) level was normalized and the IGF-1 concentration was increased with high-dose prednisone monotherapy (30 mg/d [0.50 mg/kg x d]) or rhGH (2.6 mg/d [0.043 mg/kg x d]). Prednisone monotherapy partially suppressed big IGF-2 secretion, and rhGH monotherapy acted on IGFBP-3 and ALS concentrations. FPG level was normalized with combined low-dose prednisone and rhGH, and this combination was more effective than high-dose monotherapy with either drug in reestablishing the IGF system. No adverse effects (AEs) were found. CONCLUSIONS: In this patient with inoperable NICTH, the combination of low doses of prednisone and rhGH was a successful long-term therapy for hypoglycemia, with no AEs. This therapy could be proposed for use in patients with inoperable NICTH. PMID- 15811490 TI - Sound public health policy: Focusing on saving the most lives possible. PMID- 15811489 TI - How to ensure clinical trial sensitivity in psychotropic drug development: a case study. PMID- 15811491 TI - Facility-level variation in antibiotic prescriptions for veterans with upper respiratory infections. AB - BACKGROUND: Programs have targeted individual patient and physician behaviors to reduce the use of antibiotics for upper respiratory infections (URIs), but such efforts have had limited success to date. OBJECTIVE: The aim of this study was to measure the extent of variation in antibiotic prescribing patterns at the hospital-facility level to determine whether organizational factors may be associated with patterns of antibiotic prescribing. METHODS: This was a cross sectional study using linked pharmacy and encounter data to measure hospital level variation in patterns of antibiotic prescribing at US Department of Veterans Affairs (VA) medical centers between October 1, 2000, and September 30, 2001. The main outcome measure was the proportion of visits for URIs or acute bronchitis with an antibiotic dispensed within 1 day before to 3 days after the encounter, restricted to primary-care and emergency/urgent care clinics at VA medical centers with > or =100 annual visits for URIs. RESULTS: A median of 523 visits for URIs occurred across 108 medical centers. The median proportion of visits with an antibiotic dispensed was 52% (range, 14%-88%). Hospitals in the South had increased odds of prescribing antibiotics for veterans with URIs compared with hospitals in the Northeast (odds ratio, 1.8 [95% CI, 1.2-2.5]). Among facilities with <200,000 visits per year, an increase in the percentage of unscheduled outpatient visits increased the odds of prescribing antibiotics for veterans with URIs (odds ratio per 10% increase, 1.3 [95% CI, 1.1-1.5]). CONCLUSIONS: Our results suggest variation in antibiotic prescribing for URIs at the hospital-facility level within the VA health care system. Organizational factors, such as time pressure, may be important targets for future interventions designed to reduce inappropriate antibiotic use in ambulatory care settings. PMID- 15811492 TI - Assessment of compliance with antipsychotic treatment and resource utilization in a Medicaid population. AB - BACKGROUND: Partial compliance with mental health medications has been associated with an increased risk of clinical worsening, relapse, and repeat hospitalization. OBJECTIVE: The purpose of this study was to evaluate the effect of partial compliance of patients (diagnosed as having schizophrenia or bipolar disorder) with prescribed oral atypical and conventional antipsychotic agents and the corresponding impact on resource utilization. METHODS: Patients receiving antipsychotic agents ina large Southeastern Medicaid program were grouped according to their level of compliance. Compliance was measured by the continuous, multiple interval medications available methodology. Patients were deemed partially compliant if compliance was <80%, compliant if compliance was 80% to 125%, and overly compliant if compliance was >125%. Medical costs were modeled as a function of compliance while controlling for background covariates. Logistic regression was used to model the probability of specific resource utilization. RESULTS: A total of 7864 patients were included in this analysis. After controlling for background covariates, partially compliant patients were 49.0% (95% CI, 29.2%-71.7%) more likely than compliant patients to have an inpatient hospitalization and incurred 54.5% (P < 0.001) higher inpatient charges. Partially compliant patients were also 64% (P < 0.01) more likely than compliant patients to switch or augment therapy. CONCLUSION: Partial compliance was associated with an increased risk of hospitalization and switching or augmentation of therapy when compared with being compliant. PMID- 15811494 TI - Maternal adversity, glucocorticoids and programming of neuroendocrine function and behaviour. AB - The fetus may be exposed to increased endogenous glucocorticoid or synthetic glucocorticoid in late gestation. Approximately 7% of pregnant women in Europe and North America are treated with synthetic glucocorticoid to promote lung maturation in fetuses at risk of preterm delivery. Very little is known about the mechanisms by which synthetic glucocorticoid or prenatal stress influence neurodevelopment in the human, or whether specific time windows of increased sensitivity exist. Glucocorticoids are essential for many aspects of normal brain development, but exposure of the fetal brain to excess glucocorticoid can have life-long effects on neuroendocrine function and behaviour. Both endogenous glucocorticoid and synthetic glucocorticoid exposure have a number of rapid effects in the fetal brain, including modification of neurotransmitter systems and transcriptional machinery. Such fetal exposure permanently alters hypothalamo pituitary-adrenal (HPA) function in prepubertal, postpubertal and aging offspring, in a sex-dependent manner. Prenatal glucocorticoid manipulation also leads to modification of behaviour, brain and organ morphology, as well as altered regulation of other endocrine systems. Permanent changes in endocrine function will impact on health, since elevated cumulative exposure to endogenous glucocorticoid is linked to the premature onset of pathologies associated with aging. PMID- 15811495 TI - Developmental consequences of antenatal dexamethasone treatment in nonhuman primates. AB - Research assessing fetal exposure to dexamethasone and betamethasone in animals has raised concerns about the potential for adverse side effects following antenatal treatments, not withstanding the beneficial and desired improvement in lung function. Some of the inhibitory effects on physical growth and the long term alterations in endocrine, immune and neural physiology may reflect species differences in the fetal sensitivity of rodents and monkeys to corticosteroids or perhaps could be attributed to the higher drug doses often used in animal studies. However, since steroidal drugs can be administered for extended periods in clinical practice, and also are occasionally given in the range found to cause significant effects on the brain and immune responses of infant monkeys, the simian studies have important cautionary implications for obstetrical and pediatric practice. PMID- 15811496 TI - Antenatal maternal anxiety and stress and the neurobehavioural development of the fetus and child: links and possible mechanisms. A review. AB - A direct link between antenatal maternal mood and fetal behaviour, as observed by ultrasound from 27 to 28 weeks of gestation onwards, is well established. Moreover, 14 independent prospective studies have shown a link between antenatal maternal anxiety/stress and cognitive, behavioural, and emotional problems in the child. This link generally persisted after controlling for post-natal maternal mood and other relevant confounders in the pre- and post-natal periods. Although some inconsistencies remain, the results in general support a fetal programming hypothesis. Several gestational ages have been reported to be vulnerable to the long-term effects of antenatal anxiety/stress and different mechanisms are likely to operate at different stages. Possible underlying mechanisms are just starting to be explored. Cortisol appears to cross the placenta and thus may affect the fetus and disturb ongoing developmental processes. The development of the HPA axis, limbic system, and the prefrontal cortex are likely to be affected by antenatal maternal stress and anxiety. The magnitude of the long-term effects of antenatal maternal anxiety/stress on the child is substantial. Programs to reduce maternal stress in pregnancy are therefore warranted. PMID- 15811497 TI - High antenatal maternal anxiety is related to impulsivity during performance on cognitive tasks in 14- and 15-year-olds. AB - This study prospectively investigated the influence of antenatal maternal anxiety, measured with the State Trait Anxiety Inventory at 12-22, 23-31 and 32 40 postmenstrual weeks of pregnancy, on cognitive functioning in 57 adolescents (mean age 15 years). ANCOVAs showed effects of State anxiety at 12-22 weeks, after controlling for influences of State anxiety in later pregnancy and postnatal maternal Trait anxiety. Adolescents of high anxious pregnant women reacted impulsively in the Encoding task; they responded faster but made more errors than adolescents of low anxious women. They also scored lower on two administered WISC-R subtests. In the Stop task no differences in inhibiting ongoing responses were found between adolescents of high and low anxious pregnant women. We suspect that high maternal anxiety in the first half of pregnancy may negatively affect brain development of the fetus, reflected by impulsivity and lower WISC-R scores at 14-15 years. PMID- 15811498 TI - Stress, genes and the mechanism of programming the brain for later life. AB - Adverse conditions during early life are a risk factor for stress-related diseases such as depression and post-traumatic stress disorder (PTSD). How this long-term effect of early adversity occurs is not known, although evidence accumulates that the action of stress hormones is an important determinant. In rodents after a variety of experiences, even minor ones, during postnatal life permanent changes in emotional and neuroendocrine reactivity have been observed. Also stressful events occurring prenatally and even the pre-implantation hormonal conditions can have permanent consequences. Here we will focus on evidence obtained from (i) the blastocyst implantation during conditions of ovarian hyperstimulation, which is commonly used in the generation of transgenic mice; (ii) the stress system activity in the newborn under various conditions of maternal care; (iii) the long-term consequences of maternal separation procedures. The results clearly demonstrate that early experiences trigger immediate changes in the stress system that may permanently alter brain and behaviour. PMID- 15811499 TI - The effects of prenatal social stress on behaviour: mechanisms and function. AB - In this review, we summarise experimental animal studies on the effects of the social environment during pregnancy on the offsprings' behaviour, brain, and neuroendocrine function. We show that in non-human mammals the stability of the social environment in which the pregnant females live is critical for the offsprings' social and sexual behaviour later in life as well as for reproductive functioning, endocrine state and androgen and estrogen receptor distribution in specific parts of the brain. Based on these findings, we discuss different neuroendocrine mechanisms mediating the influence of the social environment during pregnancy on the offsprings' behaviour. We conclude that maternal steroids play a decisive role in shaping foetal brain development. However, a pituitary adrenocortical pathway need not always be involved. At least in some cases an involvement of the sympathetic adrenomedullary system seems to be possible. Concerning function, we favour the hypothesis that the behavioural effects of prenatal social stress are not necessarily the 'pathological' consequences of adverse social conditions (non-adaptive explanation). Contrarily, mothers could be adjusting their offspring to the environment in which they live during pregnancy in an efficient way (adaptive explanation). PMID- 15811500 TI - Physiological stress reactivity in human pregnancy--a review. AB - Prenatal maternal stress has been found to have long-lasting effects on the behavioral and physiological development of the offspring. These programming effects on the fetus would be physiologically mediated through heightened and/or abnormal activity of the maternal sympathetic-adrenal-medullary system (SAM) and especially of her hypothalamic-pituitary-adrenocortical axis (HPA-axis). The abnormalities in maternal physiology could be present in her basal functioning, but also in her physiological reactivity to stressors, which constitutes the topic of this paper. This article reviews studies that have used laboratory challenges to study physiological stress reactivity in pregnant women. It concentrates on stress tests designed to produce pain or discomfort, or cognitive and psychological stress, and that assess changes in blood pressure, heart rate and/or cortisol as reactivity measures. The general conclusion is that physiological stress reactivity appears to be dampened during pregnancy. Nonetheless, the physiological responses to laboratory challenges are clearly present and display enough inter-individual variability to enable the study of links between responsivity patterns, psychosocial variables, fetal behavior, pregnancy outcome and offspring development. This paper also looks into the methodological limitations present in the reviewed studies. Options for sound design of stress test protocols are discussed and recommendations for future studies are presented. These methodological points are general and can therefore also be of use for researchers studying human stress reactivity in other populations and ages. PMID- 15811501 TI - Do pregnancy and childbirth adversities predict infant crying and colic? Findings and recommendations. AB - Bouts of unexplained crying in 1- to 3-month-old infants are a common problem for parents and health services. One proposed explanation has linked the crying to preceding adversities, such as maternal stress and cigarette smoking during pregnancy and complications during childbirth. In the first part of this review, we argue that studies of these links have methodological shortcomings, and make recommendations about the safeguards needed to overcome these shortcomings. In part two, we present a study that assesses the relations between adversity indices and validated measures of crying in two separate cohorts of infants. Four indices of childbirth adversity predicted infant crying separately and cumulatively in cohort 1, but not in cohort 2. We conclude that there is a need for further research that includes replication and other safeguards. Infant crying is a highly emotional issue for many parents. Before researchers add to their burden by claiming that maternal prenatal anxiety, cigarette smoking, or labour medication, contribute to their baby's crying, we need to be sure of our grounds. PMID- 15811502 TI - Prenatal programming of behavior: a twin-study perspective. AB - This paper reviews three designs that compare co-twins in tests of prenatal programming. The first design uses co-twin comparisons to attempt within-family replications of between-family associations, reported in non-twins, between reduced fetal growth and risk for later disease or behavior disorder. These replication efforts are mixed, perhaps due to small samples of weight-discordant twins, perhaps confounded by differences in the meaning of fetal growth in twins. The second design compares monozygotic co-twins from pairs differing in their placentation, with some evidence, again constrained by sample limitations that twins sharing a single placenta are more alike in dimensions of cognitive ability and personality. A third design employs a dermatoglyphic index of bilateral asymmetry as a marker of reduced buffering from environmental noise, with positive results from MZ twins that await necessary replication in longitudinal data from non-twins. Twin comparisons, especially the comparisons of monozygotic co-twins, offer an informative and incisive approach for research on prenatal programming in humans. PMID- 15811503 TI - Maternal hormones as a tool to adjust offspring phenotype in avian species. AB - Avian eggs contain substantial amounts of maternal hormones and so provide an excellent model to study hormone-mediated maternal effects. We review this new and rapidly evolving field, taking an ecological and evolutionary approach and focusing on effects and function of maternal androgens in offspring development. Manipulation of yolk levels of androgens within the physiological range indicates that maternal androgens affect behaviour, growth, morphology, immune function and survival of the offspring, in some cases even long after fledging. Descriptive and experimental studies show systematic variation in maternal androgen deposition both within and among clutches, as well as in relation to the sex of the embryo. We discuss the potential adaptive value of maternal androgen transfer at all these three levels. We conclude that maternal androgen deposition in avian eggs provides a flexible mechanism of non-genetic inheritance, by which the mother can favour some offspring over others, and adjust their developmental trajectories to prevailing environmental conditions, producing different phenotypes. However, the literature is less consistent than often assumed and at all three levels, the functional explanations need further experimental testing. The field would greatly benefit from an analysis of the underlying physiological mechanisms. PMID- 15811504 TI - Prenatal sex hormone effects on child and adult sex-typed behavior: methods and findings. AB - There is now good evidence that human sex-typed behavior is influenced by sex hormones that are present during prenatal development, confirming studies in other mammalian species. Most of the evidence comes from clinical populations, in which prenatal hormone exposure is atypical for a person's sex, but there is increasing evidence from the normal population for the importance of prenatal hormones. In this paper, we briefly review the evidence, focusing attention on the methods used to study behavioral effects of prenatal hormones. We discuss the promises and pitfalls of various types of studies, including those using clinical populations (concentrating on those most commonly studied, congenital adrenal hyperplasia, androgen insensitivity syndrome, ablatio penis, and cloacal exstrophy), direct measures of hormones in the general population (assayed through umbilical cord blood, amniotic fluid, and maternal serum during pregnancy), and indirect measures of hormones in the general population (inferred from intrauterine position and biomarkers such as otoacoustic emissions, finger length ratios, and dermatoglyphic asymmetries). We conclude with suggestions for interpreting and conducting studies of the behavioral effects of prenatal hormones. PMID- 15811506 TI - Ligand specificity of H-NOX domains: from sGC to bacterial NO sensors. AB - Soluble guanylate cyclase (sGC) is a nitric oxide (NO) sensing hemoprotein that has been found in eukaryotes from Drosophila to humans. Prokaryotic proteins with significant homology to the heme domain of sGC have recently been identified through genomic analysis. This family of heme proteins has been named the H-NOX domain, for Heme-Nitric oxide/OXygen binding domain. The key observation from initial studies in this family is that some members, those proteins from most eukaryotes and facultative aerobic prokaryotes, bind NO in a five-coordinate heme complex, but do not bind oxygen (O(2)), the same ligand binding characteristics as sGC. H-NOX family members from obligate aerobic prokaryotes bind O(2) and NO in six-coordinate complexes, similar to the globins and other O(2)-sensing heme proteins. The molecular factors that contribute to these differences in ligand specificity, within a family of sequence related proteins, are the subject of this review. PMID- 15811507 TI - Invertebrate hemoglobins and nitric oxide: how heme pocket structure controls reactivity. AB - Hemoglobins (Hbs), generally defined as 5 or 6 coordinate heme proteins whose primary function is oxygen transport, are now recognized to occur in virtually all phyla of living organisms. Historically, study of their function focused on oxygen as a reversibly bound ligand of the ferrous form of the protein. Other diatomic ligands like carbon monoxide and nitric oxide were considered "non physiological" but useful probes of structure-function relationships in Hbs. This investigatory landscape changed dramatically in the 1980s when nitric oxide was discovered to activate a heme protein, cyclic guanylate cyclase. Later, its activation was likened to Perutz' description of Hb's allosteric properties being triggered by a ligand-dependent "out-of-plane/into-plane" movement of the heme iron. In 1996, a functional role for nitric oxide in human and mammalian Hbs was demonstrated and since that time, the interest in NO as a physiologically relevant Hb ligand has greatly increased. Concomitantly, non-oxygen binding properties of Hbs have challenged the view that Hbs arose for their oxygen storage and transport properties. In this focused review we discuss some invertebrate Hbs' functionally significant reactions with nitric oxide and how strategic positioning of a few residues in the heme pocket plays an large role in the interplay of diatomic ligands to ferrous and ferric heme iron in these proteins. PMID- 15811508 TI - Assessments of the chemistry and vasodilatory activity of nitrite with hemoglobin under physiologically relevant conditions. AB - Hypoxic vasodilation involves detection of the oxygen content of blood by a sensor, which rapidly transduces this signal into vasodilatory bioactivity. Current perspectives on the molecular mechanism of this function hold that hemoglobin (Hb) operates as both oxygen sensor and a condition-responsive NO reactor that regulates the dispensing of bioactivity through release of the NO group from the beta-cys93 S-nitroso derivative of Hb, SNO-Hb. A common path to the formation of SNO-Hb involves oxidative transfer of the NO-group from heme to thiol. We have previously reported that the reaction of nitrite with deoxy-Hb, which furnishes heme-Fe(II)NO, represents one attractive route for the formation of SNO-Hb. Recent literature, however, posits that the nitrite-reductase reaction of Hb might produce physiological vasodilatory effects through NO that evades trapping on heme-Fe(II) and may be stored before release as Fe(III)NO. In this article, we briefly review current perspectives in NO biology on the nitrite reductase reaction of Hb. We report in vitro spectroscopic (UV/Vis, EPR) studies that are difficult to reconcile with suggestions that this reaction either generates a heme-Fe(III)NO reservoir or significantly liberates NO. We further show in bioassay experiments that combinations of nitrite and deoxy-Hb--under conditions that suppress SNO-Hb formation--exhibit no direct vasodilatory activity. These results help underscore the differences between physiological, RBC-regulated, hypoxic vasodilation versus pharmacological effects of exogenous nitrite. PMID- 15811509 TI - Calcium/calmodulin-dependent nitric oxide synthase activity in the CNS of Aplysia californica: biochemical characterization and link to cGMP pathways. AB - We characterized enzymatic activity of nitric oxide synthase (NOS) in the central nervous system of Aplysia californica, a popular experimental model in cellular and system neuroscience, and provided biochemical evidence for NO-cGMP signaling in molluscs. Aplysia NOS (ApNOS) activity, determined as citrulline formation, revealed its calcium-/calmodulin-(Ca/CaM) and NADPH dependence and it was inhibited by 50% with 5mM of W7 hydrochloride (a potent Ca/CaM-dependent phosphodiesterase inhibitor). A representative set of inhibitors for mammalian NOS isoforms also suppressed NOS activity in Aplysia. Specifically, the ApNOS was inhibited by 65-92% with 500 microM of L-NAME (a competitive NOS inhibitor) whereas d-NAME at the same concentration had no effect. S-Ethylisothiourea hydrobromide (5mM), a selective inhibitor of all NOS isoforms, suppressed ApNOS by 85%, l-N6-(1-iminoethyl)lysine dihydrochloride (L-NIL, 5mM), an iNOS inhibitor, by 78% and L-thiocitrulline (5mM) (an inhibitor of nNOS and iNOS) by greater than 95%. Polyclonal antibodies raised against rat nNOS hybridized with a putative purified ApNOS (160 kDa protein) from partially purified central nervous system homogenates in Western blot studies. Consistent with other studies, the activity of soluble guanylyl cyclase was stimulated as a result of NO interaction with its heme prosthetic group. The basal levels of cGMP were estimated by radioimmunoassay to be 44.47 fmol/microg of protein. Incubation of Aplysia CNS with the NO donors DEA/NONOate (diethylammonium (Z)-1-(N,N-diethylamino) diazen-1 ium-1,2-diolate - 1mM) or S-nitroso-N-acetylpenicillamine (1mM) and simultaneous phosphodiesterase inhibition with 3-isobutyl-1-methylxanthine (1mM) prior to the assay showed a 26-80 fold increase in basal cGMP levels. Addition of ODQ (1H [1,2,4]oxadiazolo[4,3-a] quinoxaline-1-one - 1mM), a selective inhibitor of soluble guanylyl cyclase, completely abolished this effect. This confirms that NO may indeed function as a messenger in the molluscan CNS, and that cGMP acts as one of its effectors. PMID- 15811510 TI - Direct single cell determination of nitric oxide synthase related metabolites in identified nitrergic neurons. AB - The biochemical characterization of individual nitrergic (NO releasing) neurons is a non-trivial task both in vertebrate and invertebrate preparations. In spite of numerous efforts, there are limited data related to intracellular concentrations of essential metabolites involved in NO synthesis and degradation. This situation creates controversies in both identification of nitrergic neurons and the selection of reliable reporters of NOS activity in heterogeneous cell populations. We take advantage of identified neurons from the pulmonate mollusc Lymnaea stagnalis to perform direct single cell microanalysis of intracellular concentrations of the major nitric oxide synthase (NOS) related metabolites such as arginine, citrulline, argininosuccinate, NO(2)(-),and NO(3)(-). Capillary electrophoresis protocols have been developed to quantitate levels of these metabolites in single identified neurons from the buccal, cerebral, and pedal ganglia using laser-induced fluorescence and conductivity detection. The limits of detection (LODs) for arginine (Arg) and citrulline (Cit) are 84 amol (11nM) and 110 amol (15 nM), respectively, and LODs for NO(2)(-)and NO(3)(-) are <200 amol (<10nM) each. We report that intracellular concentrations of NOS related metabolites are in the millimolar range and less than 1% of a single cell is required for microchemical analysis. From four cell types tested, only the esophageal motoneuron B2 contains active NOS, and they also contain surprisingly high nitrite levels (up to 5mM) compared to other neurons tested (peptidergic B4, dopaminergic RPeD1, and serotonergic CGC). These B2 neurons also exhibit an Arg/Cit ratio susceptible to the selective NOS inhibitor l-iminoethyl-N-ornithine whereas others neurons do not even though they all may contain NOS transcripts. On the contrary, we found that absolute concentrations of other NOS related metabolites including nitrates are not reliable markers of NOS activity and demonstrate the need for multiple assays for NOS activity. PMID- 15811511 TI - Thiolate coordination to Fe(II)-porphyrin NO centers. AB - The interaction of the Fe(II)-porphyrin NO model complex [Fe(TPP)(NO)] (1, TPP=tetraphenylporphyrin) with thiophenolate ligands and tetrahydrothiophene is explored both computationally and experimentally. Complex 1 is reacted with substituted thiophenolates and the obtained six-coordinate adducts of type [Fe(TPP)(SR)(NO)](-) are investigated in solution using electron paramagnetic resonance (EPR) spectroscopy. From the obtained g values and (14)N hyperfine pattern of the NO ligand it is concluded that the interaction of the thiophenolates with the Fe(II) center is weak in comparison to the corresponding 1-methylimidazole adduct. The strength of the Fe-S bond is increased when alkylthiolates are used as evidenced by comparison with the published EPR spectra of ferrous NO adducts in cytochromes P450 and P450nor, which have an axial cysteinate ligand. These results are further evaluated by density functional (DFT) calculations. The six-coordinate model complex [Fe(P)(SMe)(NO)](-) (1-SMe; P=porphine ligand used for the calculations) has an interesting electronic structure where NO acts as a medium strong sigma donor and pi acceptor ligand. Compared to the N-donor adducts with 1-methylimidazole (1-MeIm), etc., donation from the pi(h)( *) orbital of NO to Fe(II) is reduced due to the stronger trans effect of the alkylthiolate ligand. This is reflected by the predicted longer Fe NO bond length and smaller Fe-NO force constant for 1-SMe compared to the 1-MeIm adduct. Therefore, the Fe(II)-porphyrin NO adducts with trans alkylthiolate coordination have to be described as Fe(II)-NO(radical) systems. The N-O stretching frequency of these complexes is predicted below 1600cm(-1) in agreement with the available experimental data. In addition, 1-SMe has a unique spin density distribution where Fe has a negative spin density of -0.26 from the calculations. The implications of this unusual electronic structure for the reactivity of the Fe(II)-NO alkylthiolate adducts as they occur in cytochrome P450nor are discussed. PMID- 15811512 TI - A theoretical study on the binding of O(2), NO and CO to heme proteins. AB - The hybrid density functional B3LYP is used to describe the bonding of the diatomic molecules O(2), NO and CO to ferrous heme. Three different models are used, a five-coordinated porphyrin in benzene, the myoglobin active site including the distal histidine and the binuclear center in cytochrome oxidase. The geometric and electronic structures are well described by the B3LYP functional, while experimental binding energies are more difficult to reproduce. It is found that the Cu(B) center in cytochrome oxidase has a similar effect on the binding of the diatomics as the distal histidine in myoglobin. PMID- 15811513 TI - Electronic structure of cis-Mo(P)(NO)2, where P is a porphyrin: an organometallic perspective of metalloporphyrin-NO complexes. AB - This study presents a first MO analysis of the stereochemistry of cis Mo(P)(NO)(2), where the Mo(NO)(2) unit eclipses a pair of opposite Mo-N bonds and also adopts a remarkable horseshoe-like conformation. In addition, we have uncovered a number of analogies--in terms of commonalities of metal-ligand orbital interactions--between the dinitrosylmetalloporphyrins, Fe(P)(NO)(2) and Mo(P)(NO)(2), and the two dialkylmetalloporphyrins, Ru(P)(CH(3))(2), and Zr(P)(CH(3))(2). PMID- 15811514 TI - Insights into heme-based O2 sensing from structure-function relationships in the FixL proteins. AB - FixL proteins are bacterial heme-containing signal transduction proteins responsible for sensing the O(2) concentration in the organism's environment. In Sinorhizobium meliloti FixL is a protein histidine kinase that, together with its response regulator FixJ, constitute an oxygen-sensitive switch for regulation of the organism's nitrogen fixation and microaerobic respiration genes. The O(2) sensitivity of the switch is such that it transitions during the process of symbiosis in alfalfa roots. Bradyrhizobium japonicum FixL similarly regulates microaerobic and anaerobic respiration genes during symbiosis in soybean roots. FixLs responds to low oxygen concentrations with increased autophosphorylation activity of their kinase domains. The phosphorylated FixL provides a phosphoryl group to FixJ within a FixLJ complex. The phosphorylated FixJs are transcriptionally active toward their target genes. The FixL kinase domain is inhibited when the heme in FixL is oxygenated. Kinetic and thermodynamic studies of ligand binding to both ferrous and ferric FixLs have shown a generally low affinity for ligands relative to myoglobins. These relatively low ligand affinities are attributable almost completely to diminished rates of ligand binding. The heme and its environment in liganded and unliganded FixLs have been characterized by UV-visible spectroscopy, resonance Raman spectroscopy, EXAFS, and X-ray crystallography. These studies have revealed that in the purified proteins, the heme is converted from a six-coordinate low spin state to a five coordinate high spin state upon O(2) release. Comparisons of spectroscopic and structural characteristics of deoxyFixL with oxyFixL, met-FixL-CN, FixL-CO, and FixL-NO complexes indicate that distal affects in the heme pocket are, at least in part, responsible for communicating the ligation state of the heme to the kinase domain. The mechanisms by which ligand binding events are communicated from the heme to the kinase domain involves propagation and/or amplification of the ligation-coupled conformational transitions of the heme and its immediate protein environment. More recently, time-resolved experiments examining the nonequilibrium, ligand-coupled dynamics initiated by O(2), CO, and NO photolysis from the corresponding FixL complexes have begun to shed light on the landscape of the switching coordinate. Current thinking and understanding of the mechanism for signal transduction in the FixLJ systems are discussed in the context of these physical investigations. PMID- 15811515 TI - CD4(+)CD25+ T regulatory cells in murine pregnancy. AB - Mammalian pregnancy is thought to be a state of immunological tolerance and immunological pregnancy complications may result from incomplete allo-tolerance. We reported recently a higher frequency of Th1 cytokine-producing T cells specific against paternal antigens in abortion-prone mice compared to normal pregnant mice. Since Th2 cells were shown to be not essential for normal pregnancy; alloreactive Th1 cells must be differently regulated. In this context, T regulatory cells (Treg) were proposed to play an essential role. Normal pregnant mice show an expansion of CD4(+)CD25+ and IL-10+ Treg cells at the periphery compared to non-pregnant animals. Further, we reported significantly lower frequencies of Treg in abortion-prone mice. Interestingly, CD4(+)CD25+ Treg cells from normal pregnant mice were able to prevent fetal rejection. Accordingly, down-regulated levels of Treg were also reported during human miscarriage. The putative mechanisms involved in Treg-induced tolerance in mice and humans are discussed in this review. PMID- 15811516 TI - CD4(+)CD25high regulatory T cells in human pregnancy. AB - In both rodent and human systems, there is an emerging consensus that immunoregulatory activity specific for donor alloantigens is enriched in the CD4(+)CD25+ T cell population. The absence of CD4(+)CD25+ regulatory T (Treg) cells induces severe immunodeficiency with autoimmune disease, dermatitis and fatal infections in humans and mice. CD4(+)CD25+ Treg cells play a critical role in peripheral tolerance, transplantation tolerance and maternal tolerance to the fetus. Although both human and mouse CD4(+)CD25+ Treg have potent regulatory properties, surface phenotypes of human CD4(+)CD25+ Treg cells are not exactly the same as those of mouse CD4(+)CD25+ Treg cells. Murine CD4(+)CD25+ T cells are homogenous and exhibit regulatory function. On the other hand, CD4(+)CD25high T cells are the only cells which exhibit regulatory function in humans. Humans CD4(+)CD25low cells have no ability for immunosuppression. CD4(+)CD25high T cells inhibit the immunostimulation of conventional T cells through cell-to-cell contact or immunosuppressive cytokines such as interleukin 10 and transforming growth factor-beta. As another mechanism of immunosuppression, CTLA-4 on CD4(+)CD25+ regulatory T cells up-regulate indoleamine 2,3-dioxygenase (IDO) expression in dendritic cells which play important roles for immunosuppression. Here, we review the differences between humans and mouse Treg cells and the role of CD4(+)CD25+Treg during pregnancy. PMID- 15811517 TI - Fas ligand expression by maternal decidual cells is negatively correlated with the abundance of leukocytes present at the maternal-fetal interface. AB - The abundance of leukocytes at the maternal-fetal interface could influence the fate and invasion of extravillous trophoblasts. However, the mechanism(s) involved in determining the number of leukocytes present at the maternal-fetal interface as well as the nature of the interactions between invading fetal trophoblasts, maternal leukocytes and decidual cells are not well understood. In the present studies, we examined Fas ligand (FasL)/Fas expression at the maternal fetal interface in human placental tissues of early pregnancy by immunohistochemistry. The types of cells and their localization were also characterized by specific cell markers (cytokeratin, vimentin and CD45 for trophoblast, decidual cells and leukocytes, respectively). The cells undergoing apoptosis and specific apoptotic trophoblasts were detected by TUNEL assay and M30 cytoDEATH immunostaining, respectively. Using single or double immunostaining, we found that FasL expression in decidual cells was negatively correlated with the number of Fas-expressing leukocytes in the same region. Furthermore, the density of leukocytes had an inverse relationship with the number of interstitial trophoblasts present in the same area. We observed also that extravillous trophoblasts are viable despite expressing Fas and being in close proximity to decidual cell-derived FasL. These data support our hypothesis that maternal decidual cell-derived FasL may be involved in preventing the recruitment of Fas-bearing leukocytes at the maternal-fetal interface through apoptosis induction by Fas/FasL interaction, thereby promoting trophoblast invasion. PMID- 15811518 TI - Blockade of CD80 and CD86 at the time of implantation inhibits maternal rejection to the allogeneic fetus in abortion-prone matings. AB - CD28/CTLA-4 interactions with their specific B7-ligands (CD80 and CD86) play a decisive role in antigenic and allogenic responses. Recently, experimental transplant studies demonstrated that donor-specific tolerance was achieved by blocking these interactions. However, the role of blockade of CD28/B7 costimulatory pathway in the maintenance of materno-fetal tolerance has received little attention. In the present study, abortion-prone CBA/J females mated with DBA/2 males were administered with anti-CD80 and anti-CD86 monoclonal antibodies (mAbs) on day 4 of gestation (time of murine implantation). We demonstrated that the combined use of anti-CD80 and anti-CD86 mAbs induced maternal tolerance to the fetus in the abortion-prone CBA/J mice, and displayed expansion of the maternal CD4(+)CD25+ regulatory T cell population and up-regulated expression of CTLA-4, suggesting an active mechanism of regulatory T cells in suppressing maternal rejection to the fetus. In addition, the anti-CD80/86 mAbs treatment enhanced Th2 and reduced Th1 cytokine production in mice, implying that the development of Th2 cells might contribute to maternal tolerance to her fetus. Together, these findings indicated that blocking CD80 and CD86 enhanced maternal tolerance to her fetus in mice by increasing regulatory T cell function and skewing toward a Th2 response. Our data might provide an enhanced understanding of the maternal-fetal immune relationship and be helpful in clinical trials for immunotherapy of recurrent spontaneous abortion. PMID- 15811519 TI - Macrophage density in the pregnant rat uterine cervix is modulated by mast cell degranulation. AB - The uterine cervix at term undergoes histomorphological changes that resemble an inflammatory process. The aim of this study was to better characterize these changes, describing the temporal and spatial pattern of macrophages and mast cells (MC) distribution in the uterine cervix and assessing whether both cells exert a coordinated action on angiogenesis. Macrophages and MC were identified by immunohistochemistry in cervical tissue from cycling, pregnant and postpartum rats. In order to inhibit MC degranulation, pregnant rats were injected with disodium cromoglycate. The expression of vascular endothelial growth factor (VEGF) by macrophages was also evaluated. Results showed that macrophage density increased towards parturition and declined at postpartum, whereas MC density showed an inverse pattern. Interestingly, disodium cromoglycate-treated rats showed an increased number of macrophages. VEGF expression in macrophages was detected neither in control nor in treated animals; however, a coordinated action between MC and macrophages on angiogenesis could not be excluded. The present study provides a detailed mapping of macrophage and MC densities and distribution in the rat uterine cervix. Moreover, an association between macrophages and MC along pregnancy is shown, and evidence that macrophage density in the rat cervix is modulated by MC degranulation is presented. PMID- 15811520 TI - Constitutive or induced elevated levels of L-carnitine correlate with the cytokine and cellular profile of endometriosis. AB - During the past decade, accumulated evidence indicates an association between endometriosis and an alteration of humoral and cell-mediated immunity. While the role of L-carnitine in the regulation of energy metabolism is well established, it is only recently that L-carnitine has been recognized to modify the immune response in mice after in vitro or in vivo treatment. The present study has examined whether administration of L-carnitine to young female mice alters the percentage of immune cells in peritoneal exudates and the uterus as well as the levels of IFN-gamma, TNF-alpha, IL-2, IL-4, IL-6, VEGF, GM-CSF and IGF-I in blood serum, peritoneal fluid and supernatants of uterine cultured cells as tested by immunofluorescence or ELISA techniques, respectively, leading to a pathological disorder resembling human endometriosis. The results showed that, except from infertility, L-carnitine treatment resulted in a significant increase of macrophages and to a lesser degree an increase of T-cells, while elevated levels of IFN-gamma and TNF-alpha were detected in both serum and peritoneal fluid compared to controls. Although levels of L-carnitine measured in mouse serum samples using a radioisotopic method showed an increase as compared to controls, levels of acyl-L-carnitine measured in the murine peritoneal fluid samples showed a decrease similar to that measured in peritoneal fluid samples from patients with endometriosis in stage IV of the disease. These results indicate that L carnitine administration to female mice alters the cellular and growth factor profile in the uterus and peritoneum towards a phenotypical pathology similar to that of clinical endometriosis. PMID- 15811521 TI - Cytokine gene polymorphisms and susceptibility to recurrent pregnancy loss in Iranian women. AB - Recurrent pregnancy loss (RPL) is defined as three or more sequential abortions before the 20th week of gestation. There is increasing evidence to support an immunological mechanism for the occurrence of RPL. Defective production of T helper type 2 (Th2) and/or higher production of T helper type 1 (Th1) cytokines have been reported in RPL. As cytokine gene polymorphisms may be associated with different rates of cytokine production, the aim of the present study was to investigate the bi-allelic polymorphisms in TNF-alpha -308 G-->A, TNF-beta +252 G ->A, IFN-gamma +874 A-->T genes as Th1 or pro-inflammatory factors as well as IL 4 -590 C-->T, IL-10 -592 C-->A, -819 C-->T, -1082 A-->G genes as Th2 cytokines in women with RPL compared with healthy women. A total of 139 women with RPL and 143 control women with at least two successful pregnancies were included in the study. The allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) or PCR-RFLP (restriction fragment length polymorphism) methods were used for genotyping. Results indicated a significant association between the presence of CC genotype of IL-10 -592 C-->A polymorphism and the occurrence of RPL in Iranian women (63% in women with RPL and 46% in controls; OR=0.51, 95% CI: 0.3-0.85; p<0.01). There was no significant association with other positions. It may be concluded that IL-10 polymorphism at position -592 could be a genetic factor for RPL. PMID- 15811522 TI - An oligonucleotide microarray for transcriptome analysis of Schistosoma mansoni and its application/use to investigate gender-associated gene expression. AB - Global profiling transcriptomes of parasitic helminths offers the potential to simultaneously identify co-ordinately expressed genes, novel genetic programs and uniquely utilized metabolic pathways, which together provide an extensive and new resource for vaccine and drug discovery. We have exploited this post-genomic approach to fabricate the first oligonucleotide DNA microarray for gene expression analysis of the parasitic trematode Schistosoma mansoni. A total of 17,329 S. mansoni DNA sequences were used to design a microarray consisting of 7335 parasite elements or approximately 50% of this parasite's transcriptome. Here, we describe the design of this new microarray resource and its evaluation by extending studies into gender-associated gene expression in adult schistosomes. We demonstrate a high degree of reproducibility in detecting transcriptional differences among biologically replicated experiments and the ability of the microarray to distinguish between the expression of closely related gene family members. Importantly, for issues related to sexual dimorphism, labour division, gamete production and drug target discovery, 197 transcripts demonstrated a gender-biased pattern of gene expression in the adult schistosome, greatly extending the number of sex-associated genes. These data demonstrate the power of this new resource to facilitate a greater understanding into the biological complexities of schistosome development and maturation useful for identifying novel intervention strategies. PMID- 15811523 TI - Characterization of two T. gondii CK1 isoforms. AB - Previous affinity chromatography experiments have described the unexpected binding of an isoform of casein kinase I (CK1) from Leishmania mexicana, Trypanosoma cruzi, Plasmodium falciparum and Toxoplasma gondii to an immobilized cyclin-dependent kinase (CDK) inhibitor (purvalanol B). In order to further evaluate CK1 as a potential anti-parasitic target, two T. gondii CK1 genes were cloned by PCR using primers derived from a putative CK1 gene fragment identified from a T. gondii EST database. The genes are predicted to encode a smaller polypeptide of 38 kDa (TgCK1alpha) and larger 49 kDa isoform bearing a C-terminal extension (TgCK1beta). Enzymatically active recombinant FLAG-epitope tagged TgCK1alpha and TgCK1beta enzymes were immuno-precipitated from transiently transfected T. gondii parasites. While TgCK1alpha expression was found to be cytosolic, TgCK1beta was expressed predominantly at the plasma membrane. Deletion mapping showed that the C-terminal domain of TgCK1beta confers this membrane association. Recombinant TgCK1alpha and TgCK1beta isoforms were also expressed in E. coli and biochemically characterized. A 38kDa native CK1 activity was partially purified from T. gondii tachyzoites by ion-exchange and hydrophobic interaction chromatography with biochemical and serological properties closely resembling those of recombinant TgCK1alpha. In contrast, we were not able to identify a native CK1 activity corresponding to the larger TgCK1beta 49 kDa isoform in tachyzoite lysates. Purvalanol B and the related compound aminopurvalanol A selectively inhibit TgCK1alpha, confirming the existence of potentially exploitable structural differences between host and parasite CK1 enzymes. Since the more cell-permeable aminopurvalanol also inhibits parasite growth, these results provide further impetus to investigate inhibitors of CK1 as anti-parasitic agents. PMID- 15811524 TI - Plasmodium falciparum calcineurin and its association with heat shock protein 90: mechanisms for the antimalarial activity of cyclosporin A and synergism with geldanamycin. AB - Geldanamycin (GA), an antibiotic of the ansamycin family and an inhibitor of heat shock protein 90 (Hsp90), was previously shown to inhibit the malarial parasite, Plasmodium falciparum. Here we report that cyclosporin A (CsA), an inhibitor of parasitic cyclophilin (Cyp) and protein phosphatase 2B (calcineurin, CN), acted synergistically with GA to inhibit the erythrocytic growth of the parasite. Parasitic calcineurin associated with Hsp90 in vivo, and GA inhibited the association, but CsA had no effect. In a number of CsA-resistant (CsA(R)) P. falciparum clones mutations were detected in functionally significant amino acid residues of the catalytic and regulatory subunits of calcineurin (CnA and CnB, respectively) and in two out of three parasitic cyclophilins, namely Cyp19A and Cyp19B. No mutation was detected in the third cyclophilin, Cyp24. Further analysis of the mutant CnA revealed that its protein phosphatase activity was highly CsA-resistant in vitro. Similarly, one of the mutant Cyp19A proteins was purified and found to be unable to inhibit parasitic CN in the presence of CsA. Together, these results underscore the importance of the proper assembly and function of CN in plasmodial biology and suggest that the inhibition of CN can be a potential mechanism behind the CsA-sensitivity of the malaria parasite. PMID- 15811525 TI - LANCL1, an erythrocyte protein recruited to the Maurer's clefts during Plasmodium falciparum development. AB - As the malarial parasite Plasmodium falciparum develops inside the erythrocyte, parasite-derived membrane structures, referred to as Maurer's clefts, play an important role in parasite development by delivering parasite proteins to the host cell surface, and participating in the assembly of the cytoadherence complex, essential for the pathogenesis of cerebral malaria. PfSBP1 is an integral membrane protein of the clefts, interacting with an erythrocyte cytosolic protein, identified here as the human Lantibiotic synthetase component C-like protein LANCL1. LANCL1 is specifically recruited to the surface of Maurer's clefts in P. falciparum mature blood stages. We propose that the interaction between PfSBP1 and LANCL1 is central for late steps of the parasite development to prevent premature rupture of the red blood cell membrane. PMID- 15811526 TI - ATPase activity of purified plasma membranes and digestive vacuoles from Plasmodium falciparum. AB - The ATPase activity of the human malaria parasite, Plasmodium falciparum was investigated using two experimental systems, (i) digestive vacuoles, and (ii) purified plasma membranes isolated from a chloroquine-sensitive and a chloroquine resistant strain. No correlation between the level of ATPase activity and chloroquine sensitivity could be detected. In both systems, the ATPase activity of the chloroquine-resistant and -sensitive strain was decreased in the presence of the P-glycoprotein inhibitor vanadate. Susceptibility to inhibition by vanadate together with the lack of effect of ouabain implies a P-type ATPase activity in the plasma membrane. Furthermore, the inhibition of Fac8 ATPase activity by oligomycin both in the digestive vacuoles and the plasma membranes would be consistent with higher levels of Pgh1 in Fac8. Our data are consistent with the presence of a V-type H+-ATPase in the parasite food vacuole. Bafilomycin A1 and N-ethylmaleimide decreased the vacuolar ATPase activity in both chloroquine-resistant and -sensitive strains. Interestingly, a 30% decrease was observed between the ATPase activity of plasma membranes isolated from Fac8 and D10 in the presence of bafilomycin A1, suggesting the presence of a V-type ATPase in D10 plasma membrane that is underexpressed or altered in the plasma membrane of the chloroquine-resistant Fac8. The chemosensitisers tested had no effect on the ATPase activity of chloroquine-resistant P. falciparum in both systems suggesting that their activity is not mediated through an ATP-dependent mechanism. No effect was observed on the vacuolar ATPase activity in the presence of the antimalarials tested indicating that an ATP-dependent transport has not been activated. PMID- 15811527 TI - Novobiocin induces apoptosis-like cell death in topoisomerase II over-expressing arsenite resistant Leishmania donovani. AB - Leishmaniasis affects millions of people worldwide every year. Lack of effective vaccination, co-infection with other dreaded diseases like AIDS and generation of drug resistant strains demand immediate attention into this neglected area of research. The sodium m-arsenite (NaAsO2) resistant Leishmania donovani used in this study is resistant to 20 microM NaAsO2, which shows a 13-fold increase in resistance compared with wild type. Here we report that the arsenite resistant strain of L. donovani promastigotes shows cross-resistance to novobiocin, a catalytic inhibitor of topoisomerase II, with IC50 value of 320 microg ml-1 as compared with 242 microg ml-1 for wild type L. donovani. Leishmanicidal action of novobiocin induces dose- and time-dependent increase in cell death. Treatment with IC50 of novobiocin caused morphological and biochemical changes which lead to induction of cell death exhibiting characteristic features of metazoan apoptosis. Phosphatidylserine externalization, cytochrome C release to cytoplasm, activation of caspases, oligonucleosomal DNA fragmentation and in situ labelling of condensed and fragmented nuclei in both wild type and arsenite resistant L. donovani promastigotes strongly suggest the apoptosis-like mode of cell death. Cross-resistance to novobiocin in arsenite resistant strain has been correlated to over-expression of topoisomerase II and substantiated by differential inhibition of enzyme activity in wild type and arsenite resistant L. donovani. PMID- 15811528 TI - Cysteine-protease activity elicited by Ca2+ stimulus in Plasmodium. AB - Bloodstage malaria parasites require proteolytic activity for key processes as invasion, hemoglobin degradation and merozoite escape from red blood cells (RBCs). We investigated by confocal microscopy the presence of cysteine-protease activity elicited by calcium stimulus in Plasmodium chabaudi and Plasmodium falciparum in free trophozoites or for the later parasite within RBC using fluorescence resonance energy transfer (FRET) peptides. Peptide probes access, to either free or intraerythrocytic parasites, was also tested by selecting a range of fluorescent peptides (653-3146 Da molecular mass) labeled with Abz or FITC. In the present work we show that Ca2+ stimulus elicited by treatment with either melatonin, thapsigargin, ionomicin or nigericin, promotes an increase of substrate hydrolysis, which was blocked by the specific cysteine-protease inhibitor E-64 and the intracellular Ca2+ chelator, BAPTA. When parasites were treated with cytoplasmic Ca2+ releasing compounds, a cysteine-protease was labeled in the parasite cytoplasm by the fluorescent specific irreversible inhibitor, Ethyl-Eps-Leu-Tyr-Cap-Lys(Abz)-NH2, where Ethyl-Eps is Ethyl-(2S,3S) oxirane-2,3-dicarboxylate. In summary, we demonstrate that P. chabaudi and P. falciparum have a cytoplasmic dependent cysteine-protease activity elicited by Ca2+. PMID- 15811529 TI - An aspartyl protease inhibitor of Ostertagia ostertagi: molecular cloning, analysis of stage and tissue specific expression and vaccine trial. AB - Protease inhibitors are thought to protect intestinal parasitic nematodes from their hostile proteolytic environment. In a previous study, screening of Ostertagia ostertagi cDNA libraries with local antibody probes of the abomasal lymph nodes and mucus revealed a (28 kDa) aspartyl protease inhibitor (API), which was exclusively recognised by antibodies from immune calves. Here we report the molecular characterization of Oo-API (sequence analysis, developmental expression and localization) and a vaccine trial in cattle with the native and recombinant baculo-expressed antigen. The full-length open reading frame of api encodes a protein of 28 kDa. The sequence showed 82% significant homology to an Aspin homologue from Trichostrongylus colubriformis (AA034715). The cDNA encoding the full-length sequence was cloned in a bacterial pET expression vector and the pVec 35 baculovirus vector. Polyclonal rabbit serum against the Escherichia coli expressed protein was used to develop Western Blots of extracts and ES and to localize the antigen on L3, L4 and adult worm sections. The protein was expressed in all life stages, which was confirmed by real-time polymerase chain reaction (RT-PCR), and was mainly localized in the cuticle of L3, the intestinal cells of L4, and the gut and sphincter of adult worms. Polyclonal serum was also used to affinity purify the native protein. Vaccination of calves with native Oo-API and baculovirus-expressed Oo-rbAPI in combination with QuilA resulted in no protection against Ostertagia challenge infections. PMID- 15811530 TI - A bioinformatic analysis of the RAB genes of Trypanosoma brucei. AB - RAB proteins are small GTPases with vital roles in eukaryotic intracellular transport; orthologous RABs appear to fulfil similar functions in diverse organisms. Trypanosoma brucei spp., the causative organisms of Old World trypanosomiasis of humans and domestic animals, have extremely effective endocytic and exocytic mechanisms that are likely to be involved in maintenance of infection, making study of these systems of importance. Taking advantage of the essential completion of the T. brucei genome, we have re-examined the T. brucei RABs (TbRABs) so far described and identified a total of 16. BLAST searches and phylogenetic analysis show that nine of the TbRABs can confidently be assigned as orthologues or homologues of known RAB proteins from higher eukaryotes, and four more with reasonable probability. The core endocytic pathway is probably similar in complexity to yeast, whilst the early exocytic pathway appears to be more complex than in yeast. Two of the TbRAB family (RAB23 and 28) with clear mammalian orthologues appear to be unusual, and may be involved in nuclear processes and are described in more detail in an accompanying paper. Three TbRABs appear, however, to have no close homologues and may fulfil specialised functions in this organism. The availability of a complete set of TbRABs--which includes orthologues of the RABs responsible for control of the core of the endomembrane system (i.e. RAB1, 2, 4-7 and 11)--provides a first overview of the trafficking complexity that is present within a kinetoplastid parasite. Based on these homologies we suggest a systematic nomenclature for the TbRABs to reflect their functional homologies. This information is of importance both from the perspective of understanding the evolution and diversity of eukaryotic trafficking, but also in providing a framework by which to understand protein processing, trafficking, endocytosis and other related processes in these parasites. PMID- 15811531 TI - Differentiation of Leishmania donovani in host-free system: analysis of signal perception and response. AB - Leishmania donovani are the causative agents of kala-azar in humans. They undergo a developmental program following changes in the environment, resulting in the reversible transformation between the extracellular promastigote form in the sand fly vector and the obligatory intracellular amastigote form in phagolysosomes of macrophages. A host-free differentiation system for L. donovani was used to investigate the initial process of promastigote to amastigote differentiation. Within an hour after exposing promastigotes to differentiation signal (concomitant exposure to 37 degrees C and pH 5.5), they expressed the amastigote specific A2 protein family. At 5 h they started to transform to amastigote-shaped cells, a process that was completed 7 h later. This morphological transformation occurred synchronously, while cells arrested at G1. By sequential exposure to elevated temperature (for 24 h) and then acidic pH, we found that heat was responsible for the growth arrest and acidic pH to its release and subsequent route to differentiation into amastigotes. Lastly, ethanol and Azetidine 2 carboxylic acid (a synthetic proline analog) that induced heat shock response in promastigotes were capable of replacing heat in the differentiation signal. PMID- 15811532 TI - Wolbachia endosymbiont levels in severe and mild strains of Onchocerca volvulus. AB - Epidemiological, clinical and genetic data have all suggested that the filarial parasite Onchocerca volvulus, the causative agent of onchocerciasis (or river blindness) exists as two strains in West Africa. The severe strain induces severe ocular disease in a large proportion of the infected population, while the mild strain induces little ocular disease. Although DNA probes based upon a non-coding repeat sequence family can distinguish the two strains, the underlying basis for this difference in pathogenicity is not understood. Recently, several studies have implicated products produced by the Wolbachia endosymbiotic bacterium of O. volvulus in the pathogenesis of onchocerciasis. This suggested the hypothesis that differences in the Wolbachia endosymbiont population might be responsible for the pathogenic differences noted in the two strains. To test this hypothesis, quantitative PCR assays were used to measure the amount of Wolbachia DNA per nuclear genome in a collection of well characterized samples of mild and severe strain O. volvulus. The median ratio of Wolbachia DNA to nuclear DNA was significantly greater in severe strain parasites than in mild strain parasites. These data support the hypothesis that the pathogenic differences seen in severe and mild strain O. volvulus may be a function of their relative Wolbachia burden and provide additional support to the hypothesis that Wolbachia products may play a central role in the pathogenesis of ocular onchocerciasis. PMID- 15811533 TI - Targeted disruption of maebl in Plasmodium falciparum. PMID- 15811534 TI - Expression and comparative functional characterisation of recombinant acetylcholinesterase from three species of Schistosoma. PMID- 15811535 TI - Trypanosoma cruzi invades synaptotagmin VII-deficient cells by a PI-3 kinase independent pathway. PMID- 15811536 TI - CJ-15,801, a fungal natural product, inhibits the intraerythrocytic stage of Plasmodium falciparum in vitro via an effect on pantothenic acid utilisation. PMID- 15811537 TI - Litigation and the value of tobacco companies. AB - This study determines how the recent wave of litigation affected returns and systematic risk in the tobacco industry. We test for changes in stock prices coinciding with litigation announcements using a difference-in-difference event study methodology. Unfavorable information concerning litigation reduced tobacco returns. We find a decline in systematic risk as traditionally measured by the covariance between industry returns and returns to a diversified (efficient) portfolio. This decline may imply a substantial decline in the cost of equity capital of at least 2.2% after the period of litigation, which we attribute to common cost shocks experienced by the companies participating in the Master Settlement Agreement combined with the oligopolistic structure of the industry. PMID- 15811538 TI - A theory of health investment under competing mortality risks. AB - In this paper, we present a theory of health investment when there are multiple causes of death. We analyze the optimal investment rules and the comparative statics. In particular, we examine the conditions under which a cause-specific intervention has a spillover effect. By spillover effect, we mean a price reduction in one cause-specific health investment would increase all other investments. We also study the strength of the spillover effect, which is measured in terms of cross price elasticities. We find that, while a cause specific intervention might not be wasteful, the spillover effect would not be large either. PMID- 15811539 TI - Generalized modeling approaches to risk adjustment of skewed outcomes data. AB - There are two broad classes of models used to address the econometric problems caused by skewness in data commonly encountered in health care applications: (1) transformation to deal with skewness (e.g., ordinary least square (OLS) on ln(y)); and (2) alternative weighting approaches based on exponential conditional models (ECM) and generalized linear model (GLM) approaches. In this paper, we encompass these two classes of models using the three parameter generalized Gamma (GGM) distribution, which includes several of the standard alternatives as special cases-OLS with a normal error, OLS for the log-normal, the standard Gamma and exponential with a log link, and the Weibull. Using simulation methods, we find the tests of identifying distributions to be robust. The GGM also provides a potentially more robust alternative estimator to the standard alternatives. An example using inpatient expenditures is also analyzed. PMID- 15811540 TI - Is prenatal care really ineffective? Or, is the 'devil' in the distribution? AB - Prenatal care should improve infant health, yet research frequently finds only weak effects. If there are two kinds of pregnancies, 'complicated' and 'normal' ones, then combining these pregnancies may lead prenatal care to appear ineffective. Data from the National Maternal and Infant Health Survey (NMIHS) offers compelling evidence. The standard 2SLS approach yields obviously bimodal residuals and frequently insignificant prenatal care coefficients. In contrast, estimating birth weights with a finite mixture model yields estimates revealing that prenatal care has a substantial effect on 'normal' pregnancies. Our Monte Carlo experiment confirms that ignoring even a small proportion of 'complicated' pregnancies can lead prenatal care to appear unimportant. PMID- 15811541 TI - Sensitivity to scale in contingent valuation: the importance of the budget constraint. AB - The possible insensitivity of willingness-to-pay (WTP) values to changes in scale continues to cause concern within the contingent valuation (CV) literature. Although several reasons for this phenomenon have been suggested, empirical evidence both supports and refutes insensitivity. This paper presents data that suggest that insensitivity may be significantly determined by the individual's budget constraint: the higher the proportion of income the expressed WTP represents, the greater the insensitivity of that WTP to changes in the scale of the good, irrespective of changes in underlying marginal utility. The methodological implication for CV studies in health care is outlined. PMID- 15811542 TI - International differences in the impact of doctors on health: a multilevel analysis of OECD countries. AB - This paper aims to measure whether there are country variations in the efficiency of the physician workforce in reducing various measures of mortality across 21 OECD countries over 3 decades. It utilises a multilevel modelling approach both to measure country variations in physician efficiency and to explore the determinants of these variations. The results suggested that physician numbers are an important determinant of mortality across OECD countries, and cross country heterogeneity in the effect of physician availability on health is significant. We also found that availability of advanced medical technology is an important factor intervening in this relationship. PMID- 15811543 TI - The economics of vocation or 'why is a badly paid nurse a good nurse'? AB - Given the longstanding shortage of nurses in many jurisdictions, why could not nursing wages be raised to attract more people into the profession? We tell a story in which the status of nursing as a 'vocation' implies that increasing wages reduces the average quality of applicants attracted. The underlying mechanism accords with the notion that increasing wages might attract the 'wrong sort' of people into the profession and highlights an (in)efficiency wage mechanism, particular to vocations, which makes wages sticky up wards. The analysis has implications for job design in vocation-based sectors such as nursing and teaching. PMID- 15811544 TI - Graduated driver licensing and teen traffic fatalities. AB - Over the last 8 years, nearly every state has introduced graduated driver licensing (GDL) for teens. These new licensing procedures require teen drivers to advance through distinct stages where they are subject to a variety of restrictions (e.g., adult supervision, daytime driving, passenger limits). In this study, we present evidence on whether these restrictions have been effective in reducing traffic fatalities among teens. These evaluations are based on state by-year panel data from 1992 to 2002. We assess the reliability of our basic inferences in several ways including an examination of contemporaneous data for older cohorts who were not directly affected by these policies. Our results indicate that GDL regulations reduced traffic fatalities among 15-17-year-olds by at least 5.6%. We also find that the life-saving benefits of these regulations were plausibly related to their restrictiveness. And we find no evidence that these benefits were attenuated by an increase in fatality risks during the full licensure period available to older teens. PMID- 15811545 TI - Benefits transfer of willingness to pay estimates and functions for health-risk reductions: a cross-country study. AB - This paper provides a first application of the techniques of benefits transfer to the health economics literature. These techniques seek to transfer the value of some good from one 'survey' context to a new 'policy' context so avoiding the need for new valuation surveys each time a new policy question arises. Two approaches to benefits transfer are assessed: the simple transfer of mean values and the transferral of value functions. We develop a new methodology for the latter approach in which value functions are iteratively built up from theoretical principles with transfer errors being tested each time a new variable is added. Through a novel application of advanced statistical tests we show that this approach outperforms the transferral of statistically driven Best-fit functions. The case study presented focuses upon the transfer of contingent valuation (CV) willingness to pay (WTP) estimates and associated value functions for reducing the health risks associated with solar ultraviolet (UV) exposure. Common format studies are conducted in four countries with transfers between all of these being undertaken. By calculating errors in predicted versus actual values across countries we show that, when transferring between similar contexts, simple mean-value transfers outperform more complex value function transfers (with the magnitude of the former errors being encouragingly small). However, this result is reversed when transfers are undertaken across dissimilar contexts where value functions partially adjust for these differences. In summary these findings provide support and guidance for future applications. PMID- 15811546 TI - Youth alcohol use and risky sexual behavior: evidence from underage drunk driving laws. AB - Recent research calls into question previous methods for estimating the relationship between alcohol use and risky sexual behavior among youths [Rashad, I., Kaestner, R., 2004. Teenage sex, drugs and alcohol use: problems identifying the cause of risky behaviors. Journal of Health Economics 23, 493-503]. This paper provides new evidence on this question by using reductions in heavy alcohol use among underage males induced by state adoption of very strict age-targeted "Zero Tolerance" drunk driving laws. I estimate reduced form models of the effects of Zero Tolerance laws on state gonorrhea rates by age group and race over the period 1981-2000, controlling for state and year fixed effects and state specific time trends. I find that adoption of a Zero Tolerance law was associated with a significant reduction in gonorrhea rates among 15-19-year-old white males, with no effect for slightly older males age 20-24 whose drinking behavior was unaffected by the tougher policies. I find mixed effects for white females and no significant effects for blacks. While not conclusive, these results suggest an important role for alcohol use in risky sexual behavior among young men. PMID- 15811547 TI - Comparison of immunoblotted delta opioid receptor proteins expressed in the adult rat brain and their regulation by growth hormone. AB - It has previously been suggested that exogenous growth hormone (GH) affect quality of life and higher brain functions through the endogenous opioid system. Recently, we showed that GH down-regulate 72 and 48 kDa delta opioid receptor (DOR) proteins in the adult rat cerebral cortex and cerebellum. In the present study, we found that an antiserum raised against the N-terminus of the DOR also recognizes a 36 kDa protein, not recognized by a C-terminus-directed antiserum. We aimed to investigate the identity of the 72, 48 and 36 kDa proteins and to further study the effects of GH on their expression in different brain regions. The expression was studied in hypophysectomized (Hx) and untreated normal female rats. One subgroup of Hx rats received GH as a daily subcutaneous injection for 19 days. Our data show that treatment with GH in Hx rats normalized the expression of the 72 kDa protein in the cerebral cortex, whereas no significant effect were observed for the 48 or 36 kDa proteins. However, GH significantly reduced the ratio between the 72 and 36 kDa proteins in different brain regions of Hx rats. Our data suggest that GH reduces the levels of a 72 kDa DOR that likely represents a dimeric form of a 36 kDa DOR post-translationally truncated at the C-terminus, and that altered receptor dimerization may be involved in GH induced effects in the central nervous system. PMID- 15811548 TI - Developmental regulation of activated ERK expression in the spinal cord and dorsal root ganglion of the chick embryo. AB - Mitogen-activated protein kinases (MAPKs) are involved in the intracellular pathways that respond to various extracellular signals. Extracellular signal regulated kinase (ERK) is a member of MAPKs and has various functions in neural development. However, the in vivo distribution of the activated form of ERK (p ERK) in the developing nervous system is not well understood. Here, we investigated the expression of p-ERK in the spinal cord and dorsal root ganglion (DRG) of chick embryos. In the spinal cord, p-ERK-positive cells appeared in the ventral ventricular zone on embryonic day 4 (E4). From E6 onward, they appeared in the gray matter and in the white matter, suggesting migration from the ventricular zone. A double labeling method revealed that these p-ERK-positive cells included oligodendrocyte precursors. In the dorsal horn, p-ERK-positive small cells appeared on E6. Subsequently, the positive cells in the dorsal horn increased transiently in number and then decreased markedly by E10. Motoneurons also expressed p-ERK transiently on E7. In the DRG, weak p-ERK immunoreaction appeared in the ventrolateral region on E5. From E6, the immunoreactivity became stronger and by E9 intense p-ERK-positive cells were observed throughout the DRG. These data provide a neuroanatomical framework to begin to examine the in vivo role of ERK in neural development. PMID- 15811549 TI - Neurochemistry within ventrolateral medulla and cardiovascular effects during static exercise following eNOS antagonism. AB - Nitric oxide synthase (NOS), necessary for the production of nitric oxide from l arginine, exists in three isoforms: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). We have previously demonstrated that blockade of nNOS within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulated cardiovascular responses to static exercise [Ishide, T., Nauli, S.M., Maher, T.J., Ally, A., 2003. Cardiovascular responses and neurotransmitter changes following blockade of nNOS within the ventrolateral medulla during static muscle contraction. Brain Res. 977, 80-89]. In this study, we have examined the effects of bilaterally microdialyzing a specific eNOS antagonist into the RVLM and CVLM on cardiovascular responses and glutamatergic/GABAergic neurotransmission during the exercise pressor reflex in anesthetized rats. Bilateral microdialysis of a selective eNOS antagonist, l-N(5)-(1 iminoethyl)ornithine (l-NIO; 10.0 microM) into the RVLM potentiated cardiovascular responses and increased extracellular fluid glutamate levels during a static muscle contraction. At the same time, levels of GABA within the RVLM were decreased. The cardiovascular responses and neurochemical changes to muscle contraction recovered after discontinuation of the drug. In contrast, bilateral application of the eNOS antagonist into the CVLM attenuated cardiovascular responses and glutamate concentrations during a static muscle contraction, but augmented levels of GABA. These results demonstrate that eNOS within the ventrolateral medulla plays an important role in modulating glutamate/GABAergic neurotransmission, that in turn regulates the exercise pressor reflex. The present study provides further evidence of simultaneous sympathoexcitatory and sympathoinhibitory effects of nitric oxide within the RVLM and CVLM involved in the neural control of circulation during static exercise. PMID- 15811550 TI - Unilateral lesions of mesostriatal dopaminergic pathway alters the withdrawal response of the rat hindpaw to mechanical stimulation. AB - To investigate the role mesostriatal dopamine system plays in pain processing, we examined the withdrawal response of rat hindpaws to mechanical stimulus at 1, 4, and 12 weeks after unilateral 6-hydroxydopamine (6-OHDA) lesions of the mesostriatal pathway. In all of the 6-OHDA rats examined, almost no tyrosine hydroxylase (TH) immunoreactivity was detected in the substantia nigra, ventral tegmental area, and striatum ipsilateral to 6-OHDA lesions. Alteration in the withdrawal response in this model animal was evaluated by comparing the latency of withdrawal reflex following the mechanical stimulus to the hindpaw. The latency of withdrawal response in the 6-OHDA rats was significantly reduced in the side ipsilateral to 6-OHDA lesions at all times observed, whereas that was not changed through the period observed in the contralateral side, indicating that dopamine depletion in the mesostriatal system has the influence on withdrawal response to the mechanical stimulus. These results show that the unilateral dopamine depletion causes hypersensitivity to the mechanical stimulus in the ipsilateral side, suggesting that, at least in part, dopamine in the mesostriatal system may be involved in sensory processing including pain sensation induced by mechanical stimulation. PMID- 15811551 TI - Activation of the precuneus is related to reduced reaction time in serial reaction time tasks. AB - Multiple brain areas are activated during serial reaction time (RT) tasks (SRTTs), but the part of the brain that facilitates reductions in RT remains unclear. The present study attempted to determine the brain region contributing most to improved RTs during explicit SRTTs. Subjects comprised 18 healthy volunteers who were instructed to press one of four buttons corresponding to visual stimuli as quickly as possible and with minimal errors during functional MRI. Stimuli were presented either in random order (control condition) or in a repeated six-item sequence (learning condition). Conventional analysis contrasting learning and control conditions revealed activation in the prefrontal parietal area, which shifted to motor area. Subjects with high RT reduction showed more prominent activation in the precuneus than subjects with low RT reduction. Intra-subject correlation analysis revealed that time course of precuneus activation was unrelated to time-course of RT reduction. However, inter subject correlation analysis revealed that RT changes correlate only with precuneus activation, meaning that subjects showing more prominent RT reduction revealed more prominent activation of the precuneus, which is known to play critical roles in controlling finger movements with reference to buffered memory. PMID- 15811552 TI - Expression profiling with progression of dystrophic change in dysferlin-deficient mice (SJL). AB - The SJL mouse is a model for human dysferlinopathy (limb-girdle muscular dystrophy type 2B and Miyoshi myopathy). We used cDNA microarrays to compare the expression profiles of 10,012 genes in control and SJL quadriceps femoris muscles in order to find genes involved in the degeneration and regeneration process and in dysferlin's functional network. Many genes involved in the process of muscle regeneration are observed to be up-regulated in SJL mice, including cardiac ankyrin repeated protein (CARP), Neuraminidase 2, interleukin-6, insulin-like growth factor-2 and osteopontin. We found the upregulation of S100 calcium binding proteins, neural precursor cell expressed, developmentally down-regulated gene 4-like (NEDD4L) with C2 domain, and intracellular protein traffic associated proteins (Rab6 and Rab2). These proteins have the potential to interact with dysferlin. We must reveal some other molecules which may work with dysferlin in order to clarify the pathological network of dysferlinopathy. This process may lead to future improvements in the therapy for human dysferlinopathy. PMID- 15811553 TI - Hypo-and hyperthyroidism affect the ATP, ADP and AMP hydrolysis in rat hippocampal and cortical slices. AB - The presence of severe neurological symptoms in thyroid diseases has highlighted the importance of thyroid hormones in the normal functioning of the mature brain. Since, ATP is an important excitatory neurotransmitter and adenosine acts as a neuromodulatory structure inhibiting neurotransmitters release in the central nervous system (CNS), the ectonucleotidase cascade that hydrolyzes ATP to adenosine, is also involved in the control of brain functions. Thus, we investigated the influence of hyper-and hypothyroidism on the ATP, ADP and AMP hydrolysis in hippocampal and cortical slices from adult rats. Hyperthyroidism was induced by daily injections of l-thyroxine (T4) 25 microg/100 g body weight, for 14 days. Hypothyroidism was induced by thyroidectomy and methimazole (0.05%) added to their drinking water for 14 days. Hypothyroid rats were hormonally replaced by daily injections of T4 (5 microg/100 g body weight, i.p.) for 5 days. Hyperthyroidism significantly inhibited the ATP, ADP and AMP hydrolysis in hippocampal slices. In brain cortical slices, hyperthyroidism inhibited the AMP hydrolysis. In contrast, hypothyroidism increased the ATP, ADP and AMP hydrolysis in both hippocampal and cortical slices and these effects were reverted by T4 replacement. Furthermore, hypothyroidism increased the expression of NTPDase1 and 5'-nucleotidase, whereas hyperthyroidism decreased the expression of 5' nucleotidase in hippocampus of adult rats. These findings demonstrate that thyroid disorders may influence the enzymes involved in the complete degradation of ATP to adenosine and possibly affects the responses mediated by adenine nucleotides in the CNS of adult rats. PMID- 15811554 TI - Protective effect of antioxidants on brain oxidative damage caused by proline administration. AB - We have previously demonstrated that acute and chronic hyperprolinemia induce oxidative stress in cerebral cortex of rats. In the present study, we investigated the action of Vitamins E and C on the oxidative damage elicited by acute and chronic administration of proline (Pro) in rat cerebral cortex. Results showed that treatment with Vitamins E and C prevented the alterations caused by acute and chronic administration of proline on chemiluminescence, total radical trapping antioxidant potential (TRAP) and on the activities of catalase and glutathione peroxidase. If these effects also occur in the human condition, it is possible that antioxidant administration might serve as a potential adjuvant therapy to avoid the progression of the neuropsychiatric dysfunction observed in hyperprolinemic patients. PMID- 15811555 TI - Functional expression of ABCG2 transporter in human neural stem/progenitor cells. AB - We have studied the expression, localization, and function of the ABCG2 transporter, a universal stem cell marker, at the protein level in human cultured neural stem/progenitor cells (hNSPCs) using immunoblotting, immunofluorescence, and ATPase assays. Human NSPCs were isolated from human fetal brain and propagated in vitro as neurospheres. Both the cells in neurospheres and single cells dissociated from neurospheres showed high levels of ABCG2, and about 63% of the cells in neurospheres were ABCG2-positive, similar to the proportion of nestin-positive cells, and in most cases the ABCG2 and nestin staining co localized in the same cells. Both the three-dimensional structure of single hNSPCs stained with anti-ABCG2 antibodies and an examination using a biochemical marker for the plasma membrane indicated that ABCG2 was localized to the plasma membrane of hNSPCs. The ABCG2 expressed in hNSPCs had prazosin-sensitive ATP hydrolysis activity, and the ABCG2 level was sharply down-regulated during hNSPC differentiation. All these results suggested that ABCG2, was functionally expressed in hNSPCs. ABCG2 might play a significant role in maintaining human neural stem cells in an undifferentiated state and in protecting hNSPCs from xenobiotics or other toxic substances in vivo. PMID- 15811556 TI - Region-specific modulation of electrically induced synchronous oscillations in the rat hippocampus and cerebral cortex. AB - Strong tetanization induces synchronous membrane potential oscillations (seizure like afterdischarge) in mature pyramidal cells of the hippocampal CA1 region. To investigate whether local networks in other brain regions can generate such an afterdischarge independently, we studied the inducibility of afterdischarge in individual 'isolated slices' of the rat hippocampal CA1 and CA3 regions, dentate gyrus (DG), entorhinal cortex (EC), and temporal cortex (TC) using intracellular and extracellular recordings. The strong tetanization constantly induced afterdischarges in the CA1 and CA3 pyramidal cells as well as in the EC and TC superficial principal cells. However, parameters of the afterdischarges, such as the frequency and duration of afterdischarges, varied among the regions. A mixture of N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists or a GABA(A) receptor antagonist completely blocked the afterdischarges. Local GABA application during the afterdischarge elicited depolarization, rather than hyperpolarization. Moreover, reversal potentials of the afterdischarge were around -40 mV. In contrast, the tetanization resulted in occasional afterdischarge-like activities in DG slices, which were blocked by the non-NMDA or GABA(A) receptor antagonist. These findings suggest that the afterdischarges mediated through the excitatory GABAergic and glutamatergic transmissions might be common to, but be modulated differently by individual local networks in the hippocampus and cortex. PMID- 15811557 TI - Origins of lateral hypothalamic afferents associated with N-methyl-d-aspartic acid-elicited eating studied using reverse microdialysis of NMDA and Fluorogold. AB - Afferent projections to the tuberal lateral hypothalamus (tLH), where excitatory amino acid application is most effective in eliciting feeding, and to the anterior, posterior and medial regions of the hypothalamus were studied using reverse microdialysis of N-methyl-D-aspartic acid (NMDA) and Fluorogold (FG). NMDA at 660 microM delivered for 10 min was effective in stimulating food intake only when administered into the tLH, causing a mean intake of 9.3 g compared to less than 1 g in any other site. Subsequent administration of FG through the dialysis probe retrogradely in labeled neurons in brain structures associated with the feeding response including the frontal cortex, amygdala, nucleus accumbens (NA), preoptic areas, substantia nigra, ventral tegmental area (VTA), parabrachial nucleus, and the nucleus of the solitary tract (NST). Labeling after anterior and posterior LH infusion of FG was similar to that seen after tLH delivery with some apparent differences, whereas FG administration into the medial hypothalamus produced a distinctly different pattern of labeling compared to the other groups. Some of the observed labeling appeared to be almost exclusively associated with the tLH where NMDA elicits feeding. In particular, amygdala, preoptic area and shell of the accumbens labeling was noticeably denser in tLH eaters than in all other groups. These findings are consistent with the role of LH glutamate and NMDA receptors in the regulation of food intake and identify afferents to the region which possibly mediate endogenous LH glutamate's effects on feeding. PMID- 15811558 TI - Glutamic acid decarboxylase isoforms are differentially distributed in the septal region of the rat. AB - The septal region of the brain consists of a heterogeneous population of GABAergic neurons that play an important role in the generation of hippocampal theta rhythms. While GABAergic neurons employ two isoforms of the enzyme glutamic acid decarboxylase (GAD) for the synthesis of GABA, distribution of GAD isoforms has not been investigated in the septum. Immunohistochemical techniques were used to investigate the expression of GAD enzymes in medial and lateral septum. GAD65 and GAD67 immunohistochemistry revealed dense fibers and punctuated immunoreactivity in septal regions. While few GAD65-positive neuronal somas were detected in medial septum, a significantly higher number of immunoreactive neurons were detected in lateral septum. GAD65- and GAD67-positive neurons in the lateral septum exhibit higher complexity of dendritic arborizations than in the medial septum where staining was mainly restricted to the soma. Presumptive axon terminals (puncta) showed abundant immunoreactivity predominantly for GAD65 isoforms in all septal regions. This suggests that septal GABAergic neurons differentially express GAD enzymes thereby potentially reflecting functional differences. Differences found between medial and lateral septal GABAergic neuronal populations are in agreement with the concept that medial and lateral septum are brain structures with highly different connectivity and function despite anatomical proximity. PMID- 15811559 TI - The estrogen receptor (ER) alpha, but not ER beta, gene is expressed in hypothalamic growth hormone-releasing hormone neurons of the adult female rat. AB - Growth hormone (GH) synthesis and release from pituitary somatotropes is controlled by the opposing actions of the hypothalamic neuropeptides, GH releasing hormone (GHRH) in the arcuate nucleus (ARC), and somatostatin in the periventricular nucleus (PeV) and ARC. There is a striking sex difference in the pattern of GH secretion in rats. We have previously demonstrated in male rats that 70% of GHRH neurons in the ARC contain the estrogen receptor alpha (ER alpha) gene, whereas less than 5% of somatostatin neurons in the ARC and PeV expressed the ER alpha or ER beta gene. In addition, it has been reported that the PeV somatostatin neurons of neither sex possess ER immunoreactivity. However, there is no available data about colocalization of ERs and GHRH and/or somatostatin in the ARC of female rats. In this study, we used in situ hybridization in the adult female rat brain to determine whether GHRH neurons and/or somatostatin neurons in the ARC coexpress the ER alpha or ER beta gene. In the ARC, ER alpha mRNA was seen in the ventrolateral region where GHRH mRNA signals were also observed, and in the dorsomedial region where somatostatin mRNA signals were also observed. From studies using adjacent sections through these areas, the distribution of these cells appeared to overlap in part with that of cells containing ER alpha mRNA. On the other hand, few positive cells for ER beta mRNA were observed in the ARC. The double-label in situ hybridization studies showed that in the ARC, 73.4% of GHRH neurons contain ER alpha mRNA, whereas less than 5% of somatostatin neurons express the ER alpha gene. These results indicated that the majority of the GHRH neurons in ARC have ER alpha, but not ER beta, and few somatostatin neurons in ARC have ER alpha or ER beta in either adult female or male rats, suggesting that colocalization with ERs in GHRH and/or somatostatin neurons is not an important determinant of the gender specific pattern of GH secretion. PMID- 15811560 TI - Effect of Sanguis draxonis (a Chinese traditional herb) on the formation of insulin resistance in rats. AB - Sanguis draxonis (SD) is a Chinese traditional herb that is prescribed for the handling of diabetic disorders. In this study, the effects of an oral administration of SD at dosages of 100, 300, and 500 mg kg(-1) once a day, respectively, on the formation of insulin resistance were investigated in vivo in two models of insulin-resistant rats, HFD rats (high-fat diet-induced insulin resistant rats) and IILI rats (induced by the intraperitoneal injections of long acting insulin at dosage of 0.5 U kg(-1) three times daily). The insulin resistance was indicated using the loss of tolbutamide-induced hypoglycemic activity. After the oral administration of SD (300 and 500 mg kg(-1) once a day for 7 days) to HFD rats, both plasma glucose and insulin concentration were decreased significantly, while the hypoglycemic activity of tolbutamide (10 mg kg(-1), i.p.) was significantly enhanced as compared with that of the vehicle treatment (0.9% saline solution used as vehicle to disperse SD, w/v). Moreover, the formation of insulin resistance in IILI rats had been improved significantly with SD treatment (100, 300, 500 mg kg(-1) once a day for 14 days), but the influence of SD treatment on both plasma glucose and insulin concentration was not observed. For STZ-induced diabetic rats, the action of SD (300 and 500 mg kg( 1) once a day for 14 days) showed more effective on an increase of response to the exogenous short-acting porcine insulin than that of the metformin administrated orally at dosage of 320 mg kg(-1) three times daily. The present studies suggest that an oral administration of SD can increase insulin sensitivity and improve the development of insulin resistance in rats. PMID- 15811561 TI - Pressure response to carotid occlusion in diabetic rats: effect of insulin therapy. AB - Bilateral carotid occlusion (BCO) in conscious rats has been used as a maneuver to increase the sympathetic drive, producing a hypertensive response characterized by two components: an initial peak, and a maintained response of lower intensity. Acute (10-15 days) or chronic (6-13 weeks) diabetes was induced in Wistar rats with streptozotocin (STZ, 50 mg/kg, i.v.) while time-control rats received vehicle. Insulin (9 IU/kg, s.c.) was applied daily to other diabetic groups. Blood glucose was monitored three days after the administration of STZ and immediately before the experiment. Blood glucose was elevated in diabetic rats, but normal in time-control or diabetic rats treated with insulin. Basal mean arterial pressure (MAP) was reduced in diabetic as compared to time-control rats. The initial peak of the hypertensive response to BCO was blunted in either acute or chronic diabetic rats, whereas the maintained response was unaffected. Treatment of diabetic rats with insulin prevented the decrease in basal MAP and the attenuation of the initial peak caused by BCO. The maintained response was similar to that of time-control or non-treated rats. These findings suggest an abnormality of the carotid afference of the baroreflex caused by chronic hyperglycemia, which was prevented by treatment with insulin. PMID- 15811562 TI - Lifestyle modification improves risk factors in type 2 diabetes relatives. AB - AIMS: To investigate the short-term (16 weeks) effect of lifestyle intervention on insulin sensitivity, anthropometric and metabolic variables in non-diabetic first-degree relatives of type 2 diabetic patients (FDR). METHODS: Seventy-seven (49 male, 28 female) FDR were allocated to one of three groups, diet (D-group; n = 25), diet and exercise (DE-group; n = 30) or control group (C-group; n = 22). Lifestyle counselling was based on current nutrition recommendations, including increased intake of fatty fish and low glycaemic index foods. Group counselling was given on two occasions with follow-up through telephone interviews every 10 days. Assessments included insulin sensitivity index (Si), anthropometry, lipid parameters, circulating leptin and adiponectin levels. RESULTS: The D-group reduced total cholesterol (-0.31 mmol/l, P = 0.024), LDL cholesterol (-0.22 mmol/l, P = 0.021) and apolipoprotein B (-9.5 mg/dl, P = 0.009) levels, whereas the DE-group decreased body weight (-2.1%, P = 0.030) and waist circumference ( 3.0 cm, P < 0.001) versus controls. A 13% reduction in fasting insulin was observed in the DE-group, but no significant improvement in Si in D-group or DE group was observed. A subgroup, adherent to diet and who increased exercise, significantly improved Si and lipid profile. CONCLUSIONS: The improved metabolic risk profile in FDR suggests that lifestyle changes can be effective in individuals at high risk to develop type 2 diabetes and cardiovascular disease. PMID- 15811563 TI - Short-term oral ingestion of Ginkgo biloba extract (EGb 761) reduces malondialdehyde levels in washed platelets of type 2 diabetic subjects. AB - We have recently reported that ingestion of Ginkgo biloba extract (EGb 761) (a) significantly reduced collagen-induced platelet aggregation and thromboxane B2 (TXB2) production in both non-diabetic individuals as well as those with type 2 diabetes mellitus (T2DM), (b) significantly reduced platelet malondialdehyde (MDA), an index of lipid peroxidation, in non-diabetic subjects. In the present study we report that ingestion of EGb 761 (120 mg daily for 3 months), significantly decreased platelet MDA-thiobarbituric acid reacting substances (TBARS) (41 +/- 9 pmol/10(7) platelets versus 30 +/- 11 pmol/10(7) platelets) (p < 0.005) in T2DM subjects with normal cholesterol levels (total cholesterol, 164 +/- 22 mg/dl; age, 54 +/- 9 years; BMI, 35.0 +/- 8.8 kg/m2, n = 12). In T2DM subjects with high cholesterol (total cholesterol, 218 +/- 15 mg/dl; age, 52 +/- 5 years; BMI, 36.2 +/- 6.6 kg/m2, n = 7), EGb 761 ingestion reduced the platelet TBARS from 29 +/- 9 to 22 +/- 9 pmol/10(7) platelets (p < 0.04). Because ingestion of EGb 761 did not alter platelet counts it is concluded that EGb 761, probably due to the flavonoid fraction, reduced the TBARS by inhibiting cyclooxygenase (COX)-1-mediated arachidonic acid oxygenation or by reducing the arachidonic acid pool. This is likely to lead to a reduction of platelet hyperactivity, a significant contributor to the development of cardiovascular disease in T2DM patients. Because of other reported beneficial properties of EGb 761, such as stimulation of pancreatic beta-cell function in T2DM subjects with pancreatic exhaustion, it appears that T2DM subjects might benefit from ingesting EGb 761 as a dietary supplement. PMID- 15811564 TI - The haplotypes of the IRS-2 gene affect insulin sensitivity in Japanese patients with type 2 diabetes. AB - A commonly occurring nucleotide polymorphism of the insulin-receptor substrate 2 (IRS-2) gene at amino acid 1057 from Glycine to Asparaginic acid (G1057D) was recently shown to be a determinant of insulin sensitivity in both glucose tolerant individuals and those with type 2 diabetes. With respect to the latter, the IRS-2 D1057 allele increase the risk of insulin resistance among obese individuals. After we reconstructed haplotypes from the G1057D variant and the 769C/T replacement that was newly identified, we investigated the possibility that the IRS-2 gene affects insulin sensitivity in Japanese glucose-tolerant subjects (n = 260) and type 2 diabetic patients (n = 123). We did not find that the D1057 allele and haplotype pairs were associated with the risk of diabetes. However, type 2 diabetic patients, particularly obese patients, carrying the D1057 allele and the CA haplotype were associated with insulin resistance. Furthermore, we suggested that the TG and CG haplotypes might have a protective role against insulin resistance. This observation raises the possibility that both the IRS-2 D1057 allele and the CA haplotype are useful genetic markers for identifying obese individuals who are particularly susceptible to insulin resistance. PMID- 15811565 TI - Clinical characteristics at onset of Type 1 diabetes in children diagnosed between 1977 and 2001 in the south-east region of Sweden. AB - To survey clinical characteristics at diagnosis for children diagnosed with Type 1 diabetes during 25 years in the south-east part of Sweden we included all 1903 children < 16 years of age and who had been diagnosed between 1977 and 2001 in the south-east region of Sweden. A nurse or doctor in the diabetes team obtained information from medical records. Over the 25 years the mean duration of symptoms prior to diagnosis was 17.8 +/- 26.4 days and the mean glucose level at diagnosis was 23.6 +/- 9.7 mmol/l. Three percent of the children (n = 50) had a pH value < or = 7.1. The youngest children (0-5 years) had shorter duration of symptoms, lower blood-glucose levels and less often had ketonuria than the oldest children (11-15 years) but more often suffered from infections prior to diagnosis. The proportion of children diagnosed in the group 0-5 years of age increased over the study-period, apart from the last 5 years, while children with pH value < or = 7.3 decreased significantly as did the proportion of children with ketonuria or infection. The clinical characteristics at diagnosis of diabetes are heterogeneous, especially in the oldest age group. Some characteristics varied with time. PMID- 15811566 TI - Frequency and perceived burden of diabetes self-management activities in employees with insulin-treated diabetes: relationships with health outcomes. AB - We explored the relationship between frequency and perceived burden of different self-management activities and HbA1c%, symptoms of diabetes, fatigue, depression, and quality of life in 292 employees between 30 and 60 years of age with insulin treated diabetes. Participants completed questionnaires that assess self management and health-related variables. t-Tests were performed for type 1 (DM1) and type 2 diabetes (DM2) separately to compare the mean health scores of individuals who frequently or infrequently perform self-management activities and who do or do not perceive this as a burden. Participants frequently perform their self-management activities, particularly injection of insulin (96.1%), following dietary guidelines (70.8%) and eating regularly (65.6%). Dietary self-management is most often seen as a burden (70.4%), while injecting insulin is seen as least burdensome (12.8%). The perceived burden of self-management is more strongly related to health than the frequency of self-management. Frequency of self management especially relates to HbA1c% in DM1. People with DM2 who frequently follow the dietary guidelines have more positive health outcomes. Participants who perceive dietary self-management and injecting insulin as a burden have more negative health outcomes. Because different relationships were found between frequency and perceived burden of self-management and health indicators, both aspects should be assessed and considered separately when evaluating self management and examining patient's health. PMID- 15811567 TI - Metabolic improvement after intervention focusing on personal understanding in type 2 diabetes. AB - The aim of this study was to evaluate, whether an educational intervention, focusing on patients' personal understanding of their illness, was more effective than care given according to national guidelines for diabetes care. METHODS: An intervention group (n = 44), with type 2 diabetes was compared with a control group (n = 60), with HbA1c as the primary outcome. The intervention included ten group sessions addressing themes related to the patients' personal understanding of their illness. The diabetes nurses involved were educated in theories about illness/wellness experiences and participated in group sessions where various caring strategies related to the patients' individual needs and understanding were reflected upon. RESULTS: At 1-year follow-up the intervention group showed lower HbA1c levels (mean difference 0.94%; P < 0.001), lower triglycerides (mean difference 0.52 mmol/l; P = 0.002) and higher high-density lipoprotein (mean difference 0.15 mmol/l; P = 0.029) and treatment satisfaction than did the control group. The differences remained when adjusting for age, gender, body mass index or changed treatment during the intervention period. Within the intervention group, BMI and treatment satisfaction were also improved. CONCLUSION: The intervention, which focused on patients' personal understanding of illness, was found to be effective in terms of metabolic control and treatment satisfaction. PMID- 15811568 TI - Central pontine and extrapontine myelinolysis associated with type 2 diabetic patient with hypokalemia. AB - Central pontine myelinolysis (CPM) is a demyelinating disease of the pons often associated with the demyelination of extrapontine areas of the central nervous system. Although the etiology and pathogenesis are unclear, CPM is usually associated with hyponatremia or its rapid correction, and chronic alcoholism is also a common underlying condition. We observed a 43-year-old man with diabetes mellitus who developed central pontine and extrapontine myelinolysis with no apparent evidence of hyponatremia, serum hyperosmolality or associated rapid correction, or history of alcohol abuse. On admission, the patient was lethargic with dysarthria, dysphagia, and mild tetraparesis and his face and lower extremities were severely edematous. Laboratory examination showed normoglycemia and normonatremia, although hypokalemia, elevated HbA1c, and nephrotic syndrome were also present. Magnetic resonance imaging (MRI) revealed abnormal signal intensity in the pons, the deep layers of the cerebral cortex, and the adjacent white matter consistent with central pontine and extrapontine myelinolysis. Generalized edema was reduced by the use of diuretics and extracorporeal ultrafiltration without significant changes of serum sodium or osmolality. His consciousness level and paresis gradually improved within a few weeks. Our patient is a rare case of CPM associated with diabetes without apparent evidence of sodium or glucose imbalances. PMID- 15811569 TI - Quantitative effect of glycaemic improvement on the components of diabetic dyslipidaemia: a longitudinal study. AB - In order to assess the effect of glycaemic improvement on lipoprotein concentrations, we studied 73 type 2 diabetic subjects before (HbA1c 10.1 (6.2 16)%) and after (HbA1c 6.6 (3.8-8.0)%) glycaemic improvement. Total triglyceride and cholesterol (c), LDLc, HDLc, non-HDLc and apolipoproteins AI (apoAI) and B (apoB) were measured. Bivariate correlations and step-wise, multivariate analysis were performed to find predictors of change in the different components of diabetic dyslipidaemia. Changes in HDLc (r = -0.358, P = 0.001), apoAI (r = 0.355, P = 0.003), apoAI/apoB ratio (r = -0.333, P = 0.005), weight (r = -0.245, P = 0.046) and BMI (r = -0.253, P = 0.039) correlated with that of HbA1c, but, in multivariate analysis, only changes in HDLc, apoAI and apoAI/apoB ratio were predicted by the decrease in HbA1c. For the median observed change in HbA1c (-3.3 percentage-points), the estimated changes were +0.14 mmol/l, +0.12 g/l and +0.20 for HDLc, apoAI and apoAI/apoB ratio, respectively, accounting for 81, 92 and 80% of the observed changes. In conclusion, for the component of diabetic dyslipidaemia for which less therapeutic tools are available, glycaemic improvement is most effective. PMID- 15811570 TI - A toxicologic and dermatologic assessment of cinnamyl alcohol, cinnamaldehyde and cinnamic acid when used as fragrance ingredients. PMID- 15811571 TI - Fragrance material review on cinnamyl alcohol. AB - A toxicologic and dermatologic review of cinnamyl alcohol when used as a fragrance ingredient is presented. PMID- 15811572 TI - Fragrance material review on cinnamaldehyde. AB - A toxicologic and dermatologic review of cinnamaldehyde when used as a fragrance ingredient is presented. PMID- 15811573 TI - Fragrance material review on cinnamic acid. AB - A toxicologic and dermatologic review of cinnamic acid when used as a fragrance ingredient is presented. PMID- 15811574 TI - Dietary soy protein and isoflavones: no effect on the reproductive tract and minimal positive effect on bone resorption in the intact female Fischer 344 rat. AB - Our previous study evaluating 3 months of feeding soy protein or isoflavones (IF) to intact adult Sprague-Dawley (SD) rats showed no change in bone and the vagina but occasional extensive squamous metaplasia of the uterine glandular epithelium was observed. The current study was designed to characterize further these effects of soy protein or IF on the uterus using the Fischer 344 (F344) rat, a known high responder strain to estrogenic stimuli. Three-month-old intact F344 rats were divided into five groups and fed diets containing either casein, low or high amount of soy protein or casein with low or high amount of IF extract. Body weight, urinary deoxypyridinoline (Dpyr), bone mineral density (BMD) of the femur and lumbar, uterine wet weight and histology of the reproductive tract were evaluated. No significant difference was seen in bone parameters between control and treatment groups except for a lower Dpyr in the high soy and a higher lumbar BMD in the low soy groups. No alteration was seen in the reproductive tract of all treatment groups. Contrary to our hypothesis, the present results suggest that the uterus of the F344 strain is less sensitive to dietary soy protein and IF than that of the SD strain. PMID- 15811575 TI - The acrylamide intake via some common baby food for children in Sweden during their first year of life--an improved method for analysis of acrylamide. AB - The acrylamide levels in breast milk and the main categories of Swedish baby food products, i.e. breast milk substitute (infant formula), gruel, porridge and canned baby food, have been analysed. Furthermore, the acrylamide intake from these products by children up to one year of age has been estimated. Other kind of foods e.g. biscuits, are not included. Because of the expected low concentrations of acrylamide, a new sample extraction method for detection by liquid chromatography, tandem mass spectrometry, was developed and validated. The lower limit of quantification was 0.5 microg kg(-1) for liquid samples and 2 microg kg(-1) for other samples. The average levels found for gruel, porridge and canned baby food, all ready to eat, were 1.4, 26, and 7.8 microg/kg respectively. We found great variations in the acrylamide levels between and in different food categories, <0.5-64 microg/kg. For all breast milk samples except one the acrylamide level was below the limit of quantification (0.5 microg/kg). In three out of eight analysed samples of breast milk substitute, the acrylamide content was verified and possible to quantify. Assuming an acrylamide level of 0.25 microg/kg in breast milk, the mean acrylamide intake during the first six months for children who were exclusively breast-fed was estimated to be 0.04 microg/kg b.w./day. The mean acrylamide intake from breast milk and commercially made baby food during the whole first year varies due to the length of breast-feeding and the choice of baby food. The intake level range was estimated to be 0.04-1.2 microg/kg b.w/day. The mean intake between seven and twelve months of age was estimated to be about 0.5 microg/kg b.w./day. PMID- 15811576 TI - Plant-originated glycoprotein, G-120, inhibits the growth of MCF-7 cells and induces their apoptosis. AB - We characterized the biological function of G-120, glycoprotein isolated from the ethanol extract of the herb, Ulmus davidiana Nakai (UDN). G-120 has anti-tumor activity and significantly inhibited proliferation of MCF-7 cells, as measured by the thymidine uptake assay. In addition, MTT and trypan blue exclusion experiments showed that the G-120-mediated inhibition of DNA synthesis may be due to a cytostatic, rather than a cytotoxic effect. Further studies of DNA analysis and propidium iodide staining revealed that G-120 induces apoptosis in MCF-7 cells. Interestingly, G-120 (100 microg/ml) completely suppressed the binding of NF-kappaB to DNA and increased the cytosolic level of IkappaBalpha which prevented nuclear translocation of NF-kappaB. In addition, G-120 increased the expression of c-Jun, Fra-1, and Fra-2, but did not affect the expression of c Fos. Collectively, it is believed that G-120 exerts an important role in the induction of apoptosis, suppression of NF-kappaB activation, and induction of c Jun/Fra-1 or c-Jun/Fra-2 dimerization in MCF-7 cells. Consequently, G-120 could be considered as an anti-cancer agent, although further detailed experiments should be performed. PMID- 15811577 TI - Myriocin prevents fumonisin B1-induced sphingoid base accumulation in mice liver without ameliorating hepatotoxicity. AB - Fumonisin B(1) (FB(1)), a mycotoxin produced by Fusarium verticillioides present on corn and corn-based products, causes species- and organ-specific diseases. The hepatotoxic effects of FB(1) in mice have been closely correlated with the accumulation of free sphinganine, a marker for ceramide synthase inhibition, and reduced biosynthesis of more complex sphingolipids. It has been shown that FB(1) modulates expression of many cell signaling factors. In the current study we used myriocin, a specific inhibitor of serine palmitoyltransferase, to investigate the role of free sphinganine accumulation in FB(1)-induced hepatotoxicity and increased expression of selected signaling genes in BALB/c mice. The mice were pretreated daily with intraperitoneal injection of 1.0 mg/kg myriocin 30 min before subcutaneous injections of 2.25 mg/kg of FB(1) for 3 days. Results showed that myriocin alone was not hepatotoxic and the combination of myriocin plus FB(1) completely prevented the FB(1)-induced elevation of hepatic free sphinganine and prevented the FB(1)-induced induction of selected cell signaling genes, suggesting that accumulation of free sphinganine and/or its metabolites contribute to the FB(1)-modulation of the cell signaling factors. However, the combination of myriocin and FB(1) did not prevent FB(1)-increased concentration of plasma alanine aminotransferase and only slightly attenuated aspartate aminotransferase; it did not affect the FB(1)-induced hepatocyte apoptosis or increased cell proliferation. A longer combined treatment of myriocin and FB(1) was highly toxic. The hepatotoxic effects in mice seen in this study are most likely due to a combination of factors including accumulation of free sphinganine, depletion of more complex sphingolipids and sphingomyelin, or other unknown mechanisms. PMID- 15811578 TI - Aflatoxin occurrence in foodstuff supplied to dairy cattle and aflatoxin M1 in raw milk in the North of Parana state. AB - Moulds occur in a great variety of foods, including the concentrated and roughage destined for animal feeding, and can produce mycotoxins under certain conditions. The ingestion of mycotoxin AFB(1) by dairy cattle leads to the biotransformation of that substance, which is eliminated via milk as AFM(1), and also causes damage to human health. The present study aimed to analyze the presence of aflatoxins in foodstuff destined for dairy cattle and in the milk produced by these animals. The contamination of the foodstuff by aflatoxins happened mainly in the feeds and the silages did not present contamination by aflatoxins. Out of the 42 samples of milk analyzed, 10 (24%) were contaminated by AFM(1), and 3 (7%) were above the 0.5 microg/l limit. PMID- 15811580 TI - Introduction: neurobiological impact of environmental estrogens. AB - This paper provides an introduction to a special issue dedicated to the action of environmental estrogens on neural circuits and behavior. The problem of endocrine disrupting chemicals (EDCs), i.e. chemicals that have the capacity to interfere with the endocrine system, has gained increasing attention as it has become clear that these environmental contaminants may be active in humans, as well as in wildlife and domestic animal species. The majority of the early investigations were aimed at the discovery of the toxicological effects of the EDCs, but biomedical observations were among some of the first indications that estrogenic compounds may exert deleterious effects, even some time after exposure. The data derived from women exposed prenatally to diethylstilbesterol provided powerful evidence for long-term effects and endocrine disruption associated with selected compounds. The examination of wild animal populations exposed to industrial chemicals showed that the chemical exposure, though nonlethal, left the individual impaired or even incapable of reproducing. Among the multiple targets of the action of EDCs, several researches performed in recent years have investigated subtle modifications of the animal behaviors (reproductive, aggressive) that are likely to be related to alterations of specific neural pathways. We have, therefore, focused here on the behavioral studies as one of the more powerful tools to investigate EDCs effects on specific neural circuits. PMID- 15811581 TI - Phytoestrogens: hormonal action and brain plasticity. AB - Because of their protective effects in age-related diseases and hormone-dependent cancers, the use of phytoestrogens (isoflavones) as 'natural' remedies has gained prominence. Isoflavones are estrogen mimics that bind estrogen receptors and act like natural selective estrogen receptors modulators. However, limited data exists regarding the influence of soy-derived dietary isoflavones in brain. This brief review will address these topics and examine the influence of dietary isoflavones on sexually dimorphic hypothalamic nuclei. We have observed that altering the isoflavone content within diet significantly affects both the sexually dimorphic nucleus of the preoptic area (a structure that is larger in males than in females) and the anteroventral periventricular nucleus (a structure that is larger in females than in males). Specifically, when animals were switched from phytoestrogen-rich to a phytoestrogen-free diet the volume of the sexually dimorphic nucleus of the preoptic area was decreased in males (no alterations were detected in females). Conversely, when the anteroventral periventricular nucleus was examined, volume changes were recorded in males and females opposite to the patterns observed for the sexually dimorphic nucleus of the preoptic area. Given the practical limitations of examining the effects of dietary phytoestrogens in the human brain, it is important to establish comparative data sets to elucidate phytoestrogen's hormone action and potentially its beneficial brain health effects. PMID- 15811582 TI - Assessing the consequences of the pesticide methoxychlor: neuroendocrine and behavioral measures as indicators of biological impact of an estrogenic environmental chemical. AB - Japanese quail provide an advantageous avian model for assessing long-term biological consequences of endocrine disrupting chemicals (EDCs). These studies examined route of exposure and vulnerability to biological impact of EDCs over the life cycle in a precocial avian model, the Japanese quail. Embryonic exposure occurs with maternal deposition and methoxychlor (MXC) accumulated with maternal exposure. Egg injections of MXC or estradiol at selected stages of development impacted hypothalamic neuroendocrine systems in hatchlings and affected sexual maturation, with evidence for long-term effects on neurotransmitters and male behavior. Two-generation dietary studies were conducted to examine transgenerational effects of EDCs. Adult quail (P1) were exposed to dietary MXC (0, 0.5 and 5 ppm), with continued exposure in their offspring (F1), and control diet for all F2 chicks. Toxicological end points, including fertility, hatching success, and 14-day viability were unaffected. F1 and F2 male offspring from MXC treated pairs MXC had impaired mating behavior and altered plasma hormones. These studies confirm neuroendocrine and behavioral measures as reliable indices of exposure to an estrogenic EDC. Moreover, maternal deposition remains a primary route of EDC exposure, with potential deleterious consequences for field birds, especially precocial species that appear to be particularly sensitive to embryonic EDC exposure. PMID- 15811583 TI - Effects of endocrine modulators on sexual differentiation and reproductive function in male Japanese quail. AB - A number of environmental contaminants have been shown to interfere with the endocrine system. Many of these compounds bind to estrogen receptors, thereby potentially disrupting estrogen-regulated functions. In this paper, we review some background data on avian sexual differentiation and present some of the results from our studies on effects of estrogenic chemicals administered during sexual differentiation in the Japanese quail. Initially, our goal was to elucidate whether a decreased male sexual behavior in quail is a suitable endpoint for studying long-term effects of exposure to estrogenic compounds during sexual differentiation in ovo. We thereafter tested some environmental pollutants, suspected to act via mimicking estrogens, using the test system developed. Results from our studies on the synthetic estrogens ethinylestradiol and diethylstilbestrol, as well as the environmental pollutants bisphenol A, tetrabromobisphenol A, and o,p'-DDT are reviewed in this paper. We conclude that the Japanese quail is well suited as an animal model for studying various long term effects after embryonic exposure to estrogenic compounds. Depressed sexual behavior proved to be the most sensitive of the variables studied in males and we find this endpoint appropriate for studying effects of endocrine modulating chemicals in the adult quail following embryonic exposure. PMID- 15811584 TI - Effects of dietary phytoestrogens on core body temperature during the estrous cycle and pregnancy. AB - Phytoestrogens have received increased investigative attention due to their potential protective effects in connection to age-related diseases and hormone dependent cancers. Phytoestrogens appear to be an effective treatment during perimenopause where symptoms, such as hot flashes are reduced. However, little is known about the influence of phytoestrogens on core body temperature during various hormonal conditions. This study examined the effects of dietary phytoestrogens on core body temperature during estrous cycles or pregnancy by feeding Long-Evans rats either a diet rich in phytoestrogens (Phyto-600) versus a diet relatively low in phytoestrogens (Phyto-free). Independent of treatments, body temperature was highest at proestrus and declined during estrus and diestrus. Moreover, the consumption of the Phyto-600 diet moderately decreased body temperature during proestrus, estrus and diestrus versus Phyto-free-fed animals. During pregnancy, independent of treatments, core body temperature decreased as a function of increasing gestational length. Phyto-600-fed rats displayed significantly decreased body temperatures (by approximately 0.5 degrees C) from gestation days 6 to 19, compared to Phyto-free values. The results from this study indicate that consumption of dietary phytoestrogens alters the neuroendocrine mechanism of core body temperature regulation that may help explain, in part, the beneficial effects of phytoestrogens for hot flashes. PMID- 15811585 TI - Organizational effects of diethylstilbestrol on brain vasotocin and sexual behavior in male quail. AB - In Japanese quail, we previously described a sexual dimorphism of the parvocellular vasotocin system of the limbic region that, as the reproductive behavior, is steroid-sensitive and is organized during embryonic life by the exposure to estradiol. We verified in this study whether diethylstilbestrol, a chemical xenoestrogen, has analogous organizational effects on the vasotocin system of limbic regions and on copulatory behavior of male Japanese quail. We injected in the yolk sac of 3 day-old quail embryos diethylstilbestrol or estradiol benzoate (a treatment which suppresses male copulatory behavior in adulthood and reduces vasotocin innervation), or sesame oil (control). No further hormonal manipulations were performed after hatching. Sexual behavior was recorded in males at the age of 6 weeks. Estradiol- and diethylstilbestrol treated males exhibited a total suppression of copulatory behavior. After behavioral tests, all males were sacrificed and brain sections processed for vasotocin immunocytochemistry. Significant decrease in the density of vasotocin immunoreactivity was detected in the medial preoptic nucleus, in the bed nucleus of stria terminalis, and in the lateral septum of diethylstilbestrol-treated males. The magnocellular vasotocin neurons were, in contrast, not affected. In conclusion, the present data demonstrate that embryonic treatment with diethylstilbestrol induces a full sex reversal of behavioral phenotype as well as a significant decrease of vasotocin expression in the preoptic-limbic region in male Japanese quail. Therefore, the parvocellular vasotocin system could represent an optimal model to investigate the effects of pollutants on neural circuits controlling reproductive functions. PMID- 15811586 TI - D-amphetamine-related reinforcing effects are reduced in mice exposed prenatally to estrogenic endocrine disruptors. AB - Estrogenic endocrine disruptors are hormonally active compounds that can bind to estradiol receptors. Central dopamine pathways have been reported to be affected by early developmental exposure to estrogenic endocrine disruptors. In the present study, pregnant female CD-1 mice were allowed to drink spontaneously either oil or environmentally relevant low doses of two estrogenic compounds, methoxychlor (20 microg/kg) or bisphenol-A (10 microg/kg) during gestation days 11-18. Their adult offspring were assessed for conditioned place preference produced by D-amphetamine (0, 1 or 2 mg/kg). Interestingly, prenatal treatment effects were sex-dependent and no changes in conditioned place preference emerged for the male offspring. Conversely, a clear-cut profile of D-amphetamine-induced conditioned place preference was only shown by oil-exposed females, whereas exposure to bisphenol-A or methoxychlor resulted in little or no place conditioning. Locomotor effects of acute d-amphetamine were not affected by prenatal exposure to bisphenol-A or methoxychlor. As a whole, prenatal exposure to estrogenic endocrine disruptors affected some steps in the organization of the brain dopaminergic systems in the female offspring, thus leading to long-term alterations in neurobehavioral function. These data confirm that exposure to weak environmental estrogens in the period of brain sexual differentiation can influence adult behavior. PMID- 15811587 TI - Activation of brain estrogen receptors in mice lactating from mothers exposed to DDT. AB - The insecticide dichlorodiphenyltrichloroethane (DDT) interferes with physiological endocrine processes modulating estrogens receptor activity. Most of the data describing the DDT mechanism of action have been collected in vitro or in reproductive tissues in vivo. Here we use a new transgenic mouse model to investigate the DDT effects on estrogens receptor activation in vivo in non reproductive tissues. In particular, we demonstrate that DDT is able to activate estrogen receptors in the brain and the liver of adult mice after acute administration, and it is active in lactating mice when accumulated in the mother's milk. Furthermore, we demonstrate that the acute administration of DDT activates estrogen receptors with a different kinetics with respect to 17beta estradiol. Experiments with a breast cancer cell line engineered to express luciferase under the transcriptional control of activated estrogen receptors reveal that the microsomal metabolization of DDT is required for its full activity on estrogen receptors. Taken together these data lead to hypothesize that the delayed DDT time course on estrogen receptor activation in vivo might be due to a necessary step of metabolism of the compound. PMID- 15811588 TI - Chronic exposure to low doses bisphenol A interferes with pair-bonding and exploration in female Mongolian gerbils. AB - Estrogenic endocrine disruptors, synthetic or naturally occurring substances found in the environment, can interfere with the vertebrate endocrine system and, mimicking estrogens, interact with the neuroendocrine substrates of behavior. Since species vary in their sensitivity to steroids, it is of great interest to widen the range of species included in the researches on neurobehavioral effects of estrogenic endocrine disruptors. We examined socio-sexual and exploratory behavior of Mongolian gerbil females (Meriones unguiculatus), a monogamous rodent, in response to chronic exposure to the estrogenic endocrine disruptor bisphenol A. Paired females were daily administered with one of the following treatments: bisphenol A (2 or 20 microg/kg body weight/day); 17alpha-ethynil estradiol (0.04 microg/kg body weight/day 17alphaE); oil (vehicle). Females were treated for 3 weeks after pairing. Starting on day of pairing, social interactions within pairs were daily recorded. Three weeks after pairing, females were individually tested in a free exploratory paradigm. Bisphenol A and 17alphaE affected male-female social interactions by increasing social investigation. Bisphenol A reduced several exploratory parameters, indicating a decreased exploratory propensity of females. These results highlight the sensitivity of adult female gerbils to bisphenol A during the hormonally sensitive period of pair formation, also considering that the bisphenol A doses tested are well below the suggested human tolerable daily intake. PMID- 15811589 TI - Bisphenol-A exposure during pregnancy and lactation affects maternal behavior in rats. AB - In mammals, endogenous estrogens are crucial for sexual differentiation during the perinatal period, and the modulation in adulthood of many neuroendocrine and behavioral functions involved in reproduction. In rats, the estrogenic environment during pregnancy and lactation affects directly maternal behavior. This experiment was aimed to test whether the exposure to the estrogenic compound bisphenol-A (BPA; 0.040 mg/kg/die, orally) of adult female rats, from mating to weaning of the pups, could alter maternal behavior. An appropriate methodology was applied to reveal differences in the behavior of dams directed to male and female pups, testing the dams on postnatal days 3-4 and 8-9. Results show different maternal behavioral patterns towards male and female pups of control mothers, with more ano-genital licking to males than to females. Exposure of mothers to BPA modified their behavior, reducing specific components of maternal behavior, both active and passive, irrespective of the sex of pups and the period of observation. This experiment shows that maternal behavior is affected by a prolonged exposure to a low dose of BPA during pregnancy and lactation, thus suggesting an effect on neural circuits in adulthood. PMID- 15811590 TI - Early exposure to a low dose of bisphenol A affects socio-sexual behavior of juvenile female rats. AB - Play behavior is affected by alteration of the hormonal environment during development. In fact, congenital adrenal hyperplasia or early administration of diethylstilbestrol are able to modify female play behavior in mammals. In this research, play behavior of female rats was used to explore the effects of perinatal exposure to low, environmentally relevant dose of bisphenol A (BPA), a xenoestrogen widely diffused in the environment. We used 18 females born to mothers exposed to 40 microg/kg/day BPA during pregnancy and lactation, and 18 control females. The subjects were observed in a heterosexual social situation from 35 to 55 days of age. Six main behaviors were identified by principal component analysis (PCA): exploration, defensive behavior to males, play behavior with males, play behavior with females, low-intensity mating behavior, social grooming. Early administration of BPA was responsible for a significant increase of exploration (including social investigation) (p<0.001), as well as a decrease of play with males (p<0.02) and social grooming (p<0.01) at 45 days of age, indicating a general decrease of playful interactions. In general our results suggest that BPA does not induce a clear masculinization of female behavior, but is able however to defeminize some aspects of female behavior. This result is compatible with the estrogenic properties of BPA, and suggests caution in the use of a chemical that, in the range of human exposure, is able to influence the development of the brain during a critical period, resulting in long-term effects on behavior. PMID- 15811591 TI - Effects of the xenoestrogen bisphenol A in diencephalic regions of the teleost fish Coris julis occur preferentially via distinct somatostatin receptor subtypes. AB - The xenoestrogen bisphenol A, a contaminant used in the manufacturing of polymers for many consumer products, has been shown to mimic estrogenic actions. This xenoestrogen regulates secretion and expression of pituitary lactotrophs plus morphological and structural features of estrogen target tissues in rodents. Recently, ecological hazards produced by bisphenol A have drawn interests towards the effects of this environmental chemical on neurobiological functions of aquatic vertebrates of which little is known. In this study, the effects of bisphenol A on the distribution of the biologically more active somatostatin receptor subtypes in diencephalic regions of the teleost fish Coris julis were assessed using nonpeptide agonists (L-779, 976 and L-817, 818) that are highly selective for subtype(2) and subtype(5), respectively. Bisphenol A proved to be responsible for highly significant increased binding levels of subtype(2) in hypothalamic areas, while markedly decreased levels of subtype(5) were found in these diencephalic areas, as well as in the medial preglomerular nucleus. The extensive distribution of somatostatin receptor subtype(2) and subtype(5) in the teleost diencephalic areas suggests that, like in mammals, this receptor system may not only be involved in enhanced hypophysiotropic neurohormonal functions but might also promote neuroplasticity events. PMID- 15811592 TI - Neuronal inclusions in degenerative disorders Do they represent static features or a key to understand the dynamics of the disease? AB - This brief paper analyzes a few degenerative diseases expressing as movement disorders and featuring at sub-cellular level the presence of neuronal inclusions in selective brain regions. We will first draw a short draft of representative neurological diseases featuring inclusion bodies by describing the type of inclusions occurring in various disorders and analyzing both common features and distinctive aspects. As a further step, we move from the bed to the bench side discussing recent developments obtained from experimental models of these disorders which shed new light into the cause and progression of neuronal inclusions, thus helping to understand the pathophysiology of neuronal degeneration underlying movement disorders. In line with this, we will focus on recent studies which led to reproduce neuronal inclusions in vivo and in vitro by manipulating selective cellular structures/enzymatic pathways. In this way, we will try to encompass the dynamics of inclusion formation based on their fine ultrastructure and the analysis of the molecular components as well as their subcellular compartmentalization trying to relate the dynamics of inclusion formation and the pathophysiology of the disease process. An emphasis will be made on the ubiquitin proteasome system and Parkinson's disease where the analysis of neuronal inclusions enlightened potential therapeutic strategies to occlude the progression of this neuronal degeneration featured by movement disorders. PMID- 15811593 TI - The effect of Ondansetron on memory in schizophrenic patients. AB - It has been well established that patients with schizophrenia have impaired cognitive function on neuropsychological tasks related to memory. Previous studies also suggest serotonin's central role in memory. This double-blind crossover study aimed to explore the effect of Ondansetron, a selective serotonin 3 receptor (5-HT(3)) antagonist, on a variety of memory tasks in schizophrenic patients. Clozapine-treated schizophrenic patients in remission (N=21) were randomly treated with Ondansetron or placebo and then evaluated at three consecutive points. These evaluations included clinical measures (including Positive and Negative Syndrome Scale for Schizophrenia, Clinical Global Impression and Extrapyramidal Symptoms Rating Scale) and neuropsychological measures (including Digit Span, Paired Association, Rey-Osterich Complex Figure Test, Digit Symbol and the Rivermead Behavioral Memory Tests). Ondansetron, when compared with placebo, did not affect the above clinical measures and most of the neuropsychological tests. Short-term administration of Ondansetron, however, was associated with significantly improved visuo-spatial memory as measured by the Rey-Osterich Complex Figure Test. These preliminary results suggest Ondansetron's possible role in enhancement of memory function in schizophrenia. PMID- 15811594 TI - Rizatriptan does not change cerebral blood flow velocity during migraine attacks. AB - Rizatriptan represents a major advance in the treatment of migraine attack: inhibition of peripheral trigeminal nerve and constriction of intracranial extracerebral blood vessels have been proposed as its main antimigraine mechanisms of action. Although many studies may suggest that rizatriptan causes highly selective vasoconstriction within intracranial extracerebral vessels (i.e., meningeal arteries), no literature data are available to date on possible cerebral hemodynamic changes in humans after treatment with rizatriptan. The aim of this study was to evaluate the effect of rizatriptan on cerebral blood flow velocity performing transcranial Doppler during spontaneous attacks of migraine without aura. Fourteen patients suffering from migraine without aura were monitored to evaluate mean flow velocity changes on both middle cerebral arteries during migraine attack 30 min before and 120 min after oral administration of rizatriptan 10mg. Monitoring was repeated for 30 min during the pain-free period. All patients turned out to be drug responders and no significant mean flow velocity changes were observed between the pain-free period and pre-treatment phase; besides no significant difference in mean flow velocity value have been detected between the periods after the drug administration during the attack versus both pre-treatment period and pain-free phase. These findings indicate that the antimigraine action of rizatriptan is not associated with clear intracranial cerebral hemodynamic changes and may support its cerebrovascular safety. PMID- 15811595 TI - Grammatical gender in the brain: evidence from an fMRI study on Italian. AB - The grammatical gender of a word is a lexical-syntactic property determining agreement among different sentence parts. Recent fMRI investigations identified the areas involved in the retrieval of grammatical gender near the left Broca's area providing further evidence to confirm the preeminent syntactic role of this area. However, these studies employed categorical designs based on the controversial methodology of the cognitive subtraction of neural activations related to different tasks. In the present study we identified the neural substrates of grammatical gender assignment using an fMRI parametric study. Participants decided the grammatical gender of visually presented Italian words whose gender-to-ending regularity varied. The results showed activation in left and right fronto-temporal areas suggesting an interplay of both hemispheres in the processing of grammatical gender. PMID- 15811596 TI - Slow cortical potential shifts preceding sensorimotor interactions. AB - It is well known that synchronization of cortical neurons is modulated ("gating") by the chronological interaction between somatosensory and sensorimotor events. This study tested the hypothesis that the anticipatory processes for this interaction increase the synchronization of cortical neurons as revealed by negative event-related potentials (contingent negative variation, CNV). High resolution electroencephalographic data (128 electrodes) were recorded in 14 subjects. In the "sensorimotor interaction" condition, the subjects were waiting for a galvanic somatosensory stimulation at the left hand concomitant with a Go or NoGo stimulus (50% of Go trials triggering right hand movements). In the control condition, the Go/NoGo stimulus followed the somatosensory stimulation of 1.5s. The electroencephalographic data were spatially enhanced by surface Laplacian estimation. In the control condition, the CNV was observed only in the foreperiod between the somatosensory stimulation and Go/NoGo task (i.e. no CNV before the somatosensory stimuli). It was spatially localized in the primary sensorimotor area contralateral to the possible motor response. In the "sensorimotor interaction" condition, the CNV preceded the concomitant somatosensory stimulation and Go/NoGo task and was distributed to the frontocentral midline other than the contralateral sensorimotor area. These results suggest that the anticipatory processes for sensorimotor interactions increase the synchronization of cortical neurons in the frontocentral midline, possibly due to mechanisms sub-serving top-down attentional processes. PMID- 15811597 TI - Stereological analysis of microvascular parameters in a double transgenic model of Alzheimer's disease. AB - Morphological alterations in microvasculature occur as a common finding in the brains of non-demented aged persons and patients with Alzheimer's disease. Quantifying the extent of this vascular pathology, however, has been complicated by systematic error (bias) associated with the applications of assumption- and model-based morphometric techniques to human and animal tissues. The current study used novel assumption- and model-free stereological approaches to quantify capillary parameters in the corpus callosum of a double amyloid precursor protein/presenilin-1 transgenic murine model of Alzheimer's disease. The results revealed significant reductions in the total number of capillary segments in white matter of transgenic mice compared to non-transgenic littermates, with no differences in total capillary length. These findings support the view that the expression of mutant human genes for beta-amyloid peptides alters the normal architecture of cerebral capillary vessels in the white matter of mouse brain, which may model microvasculature changes reported in Alzheimer's disease. PMID- 15811598 TI - Ejaculation induces long-lasting behavioural changes in male rats in the forced swimming test: evidence for an increased sensitivity to the antidepressant desipramine. AB - Copulation to exhaustion induces a sexual inhibitory state featured by a decreased motivation. Since diminished motivation is a key symptom of depression, we analysed if sexually exhausted animals showed increased levels of depressive like behaviour in the forced swimming test (FST). Besides, sexual activity has been reported to have reinforcing properties. Thus, we analysed whether different levels of sexual activity modified the development of the behavioural despair in the FST. Finally, the effect of a sub-threshold dose of desipramine (DMI, 2.5mg/kg) was evaluated in animals with different sexual conditions. Male adult rats were divided into: (a) naive rats and (b) animals executing one intromission (1-INTR) or (c) one ejaculation (1-EJ) and (d) sexually satiated rats, classified as sexually responsive (R) and non-responsive (NR). No differences were found in immobility behaviour between sexually exhausted and naive rats. In the pre-test sessions of the FST males attaining ejaculation (1-EJ, R and NR) had lower levels of immobility or showed a tendency towards such a diminution, while animals with sexual activity not involving ejaculation did not. Data suggest a "protective" effect of ejaculation against the development of depressive-like behaviour. Finally, the sub-effective dose of DMI produced an antidepressant-like action in all animals that ejaculated. In conclusion, sexual experience involving ejaculation modifies the behavioural expression of rats in the FST and their sensitivity to antidepressant drugs like DMI. Both responses reflect brain plastic changes induced by ejaculation. PMID- 15811599 TI - Microarray analysis of rat brain gene expression after chronic administration of sodium valproate. AB - Valproic acid has been used to treat mania and bipolar disorder, but its mechanism of action is not agreed on. We used rat genome U34A Affymetrix oligonucleotide microarrays, containing 8799 known probesets, to determine the effect of 30-day daily intraperitoneal administration of valproate (200mg/kg) on rat brain gene expression. We found 87 down-regulated genes and 34 up-regulated genes of at least a 1.4-fold change in valproate-treated compared to control rats. The experiments were done on five independent samples for each group, each in duplicate. The genes affected are known to be involved in a variety of pathways, including synaptic transmission, ion channels and transport, G-protein signaling, lipid, glucose and amino-acid metabolism, transcriptional and translational regulation, phosphoinositol cycle, protein kinases and phosphatases, and apoptosis. Our results suggest that the therapeutic effect of valproate may involve the modulation of multiple signaling pathways. PMID- 15811601 TI - Quantitative spatial analysis of the distribution of NADPH-diaphorase-positive neurons in the developing and mature rat retina. AB - NADPH-diaphorase (NADPH-d) histochemistry labels a subpopulation of nitric oxide synthesizing amacrine cells in the inner nuclear layer of the rat retina. We have studied their morphology and distribution in postnatal and adult rats in whole mounted retinae. NAPDH-d-positive neurons are detected as early as postnatal day (P)5, especially in the peripheral retina; intense labeling of somata and long lengths of dendrites is obtained between P10 and P18, after which only the somata exhibit NADPH-d activity. The density and number of these cells increase progressively from P7 to P14, with a significantly higher density in the central retina as compared to the periphery. The sociology of these cells was analyzed quantitatively studying the Voronoi domains: a polygon area can be drawn that delineates the territory of the map that is closer to the cell than to any other cell of the map. In addition, we calculated the conformity ratio of Cook, i.e., the mean nearest neighbor distance/standard deviation of all the nearest neighbor distances, in order to reveal whether or not these cells are regularly distributed through the retina. We find that the distribution of the NADPH-d positive cells tends to be regular throughout the retina: the local coefficient of variation (obtained by comparing the size of each Voronoi polygon area to those of its neighbors) tends to regularity at P14 and remains unaltered through maturity. Therefore, as other cell types, NADPH-d-positive amacrine cells are almost regularly distributed from the time of eye opening and nitric oxide may play a role in the development of retinal circuitry and in the regulation of retinal blood flow. PMID- 15811600 TI - Strain differences of dopamine receptor levels and dopamine related behaviors in rats. AB - Here we have investigated whether differences in levels of dopamine D1-like, D2 like receptors, dopamine D3 receptors, and dopamine transporter could be related to behaviors such as immobility response and locomotion between Wistar rats and Sprague-Dawley rats. The levels of the dopamine receptors and transporter were measured by autoradiographic study at the level of basal ganglia and the limbic subregion. The behavioral study was done by open-field and immobility response tests. The Wistar rats exhibited a higher level of D1 receptor binding in the basal ganglia subregions than Sprague-Dawley rats. The Wistar rats have higher levels of dopamine D2 receptor binding and dopamine transporter binding in the dorsolateral part of the caudate-putamen. In addition, the dopamine transporter binding were also higher in the Wistar rats than in Sprague-Dawley rats in the ventral part of the caudate-putamen and nucleus accumbens core. However, there were no differences in the level of D3 receptor binding in the limbic or basal ganglia subregions between these two strains. In Wistar rats, the duration of the immobility responses was longer and with less locomotor activity after these immobility responses compared with Sprague-Dawley rats. These data suggest that the differences in dopamine receptors in these two rat strains may in part relate to the behavioral differences reported in these two strains. PMID- 15811602 TI - The protective role of DL-alpha-lipoic acid in biogenic amines catabolism triggered by Abeta amyloid vaccination in mice. AB - The major pathological consequence of Alzheimer disease (AD) is accumulation of beta-amyloid (Abeta) peptide fibrillar plaque in the brain and subsequent inflammatory reaction associated with the surrounding cells due to the presence of these aggregates. Inflammation is the major complication associated with Abeta peptide vaccination. Abeta peptide activated T-helper cells are shown to enhance the existing-inflammatory conditions in the brain and other organs of AD patients. Hence systematic studies on potential approaches that will prevent inflammation during the vaccination are highly desired. DL-alpha-lipoic acid (LA), an antioxidant with known function as cofactor in mitochondrial dehydrogenase reactions, will be a good candidate to annul the oxidative damage due to vaccination triggered inflammation. For the first time, levels of principal neurotransmitters and their major metabolites in hippocampus and neocortex regions of brain are quantified to find out the level of inflammation. We have used high performance liquid chromatography with electro chemical detection (HPLC-EC) for monitoring neurotransmitter levels. We have shown a significant (p<0.05) reduction of 5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE) in the systemic inflammation induced (SI), vaccinated (VA) and inflammation induced vaccinated (IV) mice. Nevertheless their metabolites such as 5-hydroxyindole acetic acid (5-HIAA) and homovanillic acid (HVA) are significantly (p<0.05) increased when compared with control. Interestingly, antioxidant LA treated mice with systemic inflammation (IL), vaccinated (VL) and inflammation induced vaccinated (IVL) mice exhibited enhanced level of 5-HT, DA and NE and the concentration of 5-HIAA and HVA gradually returned to normal. These results suggest a possible new way for monitoring and modifying the inflammation and thereby preventing Abeta vaccination mediated tissue damage. PMID- 15811603 TI - Effects of effort and EMG levels on short-latency stretch reflex modulation after varying background muscle contractions. AB - It is known that the short-latency stretch reflex (SLSR) is modulated by the background muscle activity when it is elicited at matched torque levels. This study was designed to examine the effects of muscle contraction types before a stretch perturbation on SLSR in the human soleus muscle (SOL) when SLSR was elicited at the same levels of effort and at matched electromyographic (EMG) activity levels. A mechanical stretch perturbation was applied to the calf muscles when the ankle joint reached a ninety degree tibio-tarsal joint angle after the muscles performed an isometric (pre-ISO), shortening (pre-SHO) and lengthening contraction (pre-LEN). Subjects were seated on an ankle ergometer chair and developed 0%, 10%, 20%, 30%, 40%, 50%, 60% and 70% ankle joint torque (AJT) of maximum voluntary isometric plantar flexion contraction at 80 degrees in pre-SHO, at 90 degrees in pre-ISO and at 100 degrees in pre-LEN. After that, isometric or dynamic contractions started, and the subjects were asked to maintain effort levels as, needed, to maintain the target torque levels until the end of the stretch. They relaxed their muscles fully after the stretch. This chain of processes was consecutively repeated 10 times. EMG signals obtained from SOL were averaged after they were high-pass filtered and full-wave rectified. Some major findings resulted: (1) there were no differences in SLSR area in the active muscle between pre-ISO and pre-SHO, whereas its waveform was steeper in pre-ISO than in pre-SHO. (2) SLSR p-to-p amplitude and waveform were larger and steeper in the active muscle than in the relaxed one in all conditions, whereas they were independent of the effort levels once the muscle was activated. This led to steady SLSR modulation in response to the background muscle contraction in the active muscle regardless of whether the SLSR was elicited at matched AJT or EMG activity levels. These findings suggest that SLSR is closely related to the muscle spindle sensitivity influenced by the following factors: (1) the background muscle contraction type, and (2) gamma motoneuron activity set by CNS based on the effort level. PMID- 15811604 TI - Estimate of M-wave changes in human biceps brachii during continuous stimulation. AB - The purpose of the present study was to validate the capability of new fatigue indexes (in the time and frequency domain) applied to experimental recordings and thus, to test some assumptions made in previous simulations. The indexes were applied to M-waves detected non-invasively from human m.biceps brachii during repetitive slightly above threshold stimulations. It was found that distance between the motor point and middle of the end-plate region could be relatively large. Under identical conditions (signals detected by monopolar electrodes and high-pass filtered at 1 Hz), the relative changes of the indexes obtained in electrophysiological experiments and simulations were similar. Changes of the intracellular action potential profile during fatigue used in the simulations were consequently supposed to be close to the actual ones for the muscle analyzed. When the high-pass cut-off frequency was higher than 1 Hz, the sensitivity of the index in the time domain was higher, while that in the frequency domain was lower. If the normalizing spectral moment was of higher order, the sensitivity of the spectral index could be even 150-times greater than that of the fatigue indexes traditionally used. Thus, the spectral index promises high capability to assess fatigue during functional electrical stimulation. PMID- 15811605 TI - Co-contraction during static and dynamic knee extensions in ACL deficient subjects. AB - Co-contraction of the muscles is proposed in the literature as one of the strategies that anterior cruciate ligament deficient (ACLD) subjects can use to compensate the loss of ACL function. This study examined the response of ACLD and control subjects to different shear forces in isometric and slow-dynamic knee extensions. Twelve chronic ACLD and 10 control subjects performed submaximal positioning and slow-dynamic knee extensions (between 45 degrees and 5 degrees of knee flexion) with two external flexion moments both applied at two distances on the lower leg. The shear force was controlled by changing the moment arm without changing the moment. Electromyographic data were collected from knee flexor and extensor muscles. In the analysis of variance, no significant effect of subject group was found in positioning or slow-dynamic tasks across all muscles. The effect of knee angle was significantly different between the subject groups for biceps femoris in positioning and for rectus femoris in slow-dynamic tasks, but these effects were very small and will not have a great impact on the resulting shear forces. There was no interaction between moment arm and subject group. Therefore, the hypothesis that ACLD subjects increase co-contraction in situations with an increased shear load in positioning and slow-dynamic knee extensions could not be confirmed. PMID- 15811606 TI - Prediction of handgrip forces using surface EMG of forearm muscles. AB - Evaluation of handgrip forces constitutes an essential component of ergonomic evaluation (e.g. of hand tools), but is difficult to perform at the workplace. The present study describes a series of experiments on 8 healthy male subjects to determine the validity of linear regression models using the surface electromyography (EMG) of up to 6 forearm muscles to predict handgrip forces. For isometric gripping tasks, normalized EMG to grip force calibrations using a series of dynamic force bursts up to 300 N resulted in a valid prediction of grip forces based on the EMG of 6 forearm muscles. Absolute differences between observed and predicted grip force were small (between 27 and 41 N) which shows that the proposed method might be used for the ergonomic evaluation of the use of hand tools. The EMG - handgrip force model appeared to be minimally affected by grip width, i.e. a model for 67 mm grip width was able to validly predict grip forces for 59 and 75 mm grip widths. Furthermore, it was shown that of the 6 forearm muscles studied at least 3 have to be assessed to arrive at a sufficient level of validity, while it seems to be irrelevant which 3 of those 6 forearm muscles are assessed. PMID- 15811607 TI - Force, work and power output of lower limb muscles during human maximal-effort countermovement jumping. AB - The purpose of this study was to simulate human maximal-effort countermovement jumping with a three-dimensional neuromusculoskeletal model. The specific aim was to investigate muscle force, work and power output of major lower limb muscles during the motion. A neuromusculoskeletal model that has nine rigid body segments, 20 degrees of freedom, 32 Hill-type lower limb muscles was developed. The neural activation input signal was represented by a series of step functions with step duration of 0.05 s. The excitation-contraction dynamics of the contractile element, the tissues around the joints to limit the joint range of motion, as well as the foot-ground interaction were implemented. A simulation was started from a standing posture. Optimal pattern of the activation input signal was searched through numerical optimization with a goal of maximizing the height reached by the mass center of body after jumping up. As a result, feasible kinematics, ground reaction force profile and muscle excitation profile were generated. It was found that monoarticular muscles had major contributions of mechanical work and power output, whereas biarticular muscles had minor contributions. Hip adductors, abductors and external rotator muscles were vigorously activated, although their mechanical work and power output was minor because of their limited length change during the motion. Joint flexor muscles such as m. iliopsoas, m. biceps femoris short head and m. tibialis anterior were activated in the beginning of the motion with an effect of facilitating the generation of a countermovement. PMID- 15811608 TI - Mononeuropathies of the radial nerve: clinical and neurographic findings in 91 consecutive cases. AB - Retrospective features of 91 consecutive cases (68 men, 23 women; mean age 44.4 years) of radial mononeuropathy diagnosed over the last 8 years in two electromyography (EMG) services are reported to define the clinical and electrophysiological findings of radial neuropathies in relation to traumatic and non-traumatic causes and site of injury. The occurrence of radial neuropathy was 0.65 x 100 first electromyographic examinations. The most frequent site of damage was the main trunk at the spiral groove of the humerus (36%); the most frequent cause was nerve trauma (70%) due to fracture (36%). In neuropathies of the main trunk and posterior interosseous (PI) nerve, "complete nerve injury" was observed in 36% of cases, conduction motor block in 33% and motor conduction velocity slowing in 46%. At least one of these findings was present in 51%, whereas motor neurography was normal in 13% of cases. Sensory action potential (SAP) anomalies were observed in 51% of cases. In neuropathy of the superficial radial nerve, no SAP was detected in 30% of cases; in all others except one, SAP was reduced in amplitude. Non-traumatic neuropathies showed severer conduction block and less severe anomalies of SAP than traumatic neuropathies. No differences were found between men and women. EMG is essential for confirming the site of injury and neurographic study may be helpful for diagnosis, providing information about lesion type and severity. PMID- 15811609 TI - Quantification of the dynamic properties of EMG patterns during gait. AB - A technique for analyzing and comparing the dynamic properties of electromyographic (EMG) patterns collected during gait is presented. A gait metric is computed, consisting of both magnitude (amplitude) and phase (timing) components. For the magnitude component, the processed EMG pattern is compared to a normative EMG pattern obtained under similar walking conditions, where the metric is incremented if the muscle is firing during expected active regions or is silent during expected inactive regions. The magnitude metric is penalized when the EMG is silent during phases of expected activity or when the EMG is active in regions of expected inactivity. The phase component of the metric computes the percentage of the gait cycle when the muscle is firing appropriately, that is, active in expected active regions and silent in expected inactive regions. The magnitude and phase components of the metric are normalized and combined to yield the EMG pattern that demonstrates the closest characteristics compared to normative gait data collected under similar walking conditions. Using experimental data, the proposed gait metric was tested and accurately reflects the observed changes in the EMG patterns. Clinical uses for the gait metric are discussed in relation to gait therapies, such as determining optimal gait training conditions in individuals following stroke and spinal cord injury. PMID- 15811610 TI - Quantification of level of effort at the plantarflexors and hip extensors and flexor muscles in healthy subjects walking at different cadences. AB - The plantarflexor, hip extensor and hip flexor muscle groups contribute by their concentric action to generate most of the energy during level gait in healthy subjects. The goal of the present study was to determine, during the main energy generation phases, the relative demand of these three groups in 14 healthy subjects walking at four cadences (self-selected, 60, 80 and 120 steps/min). The muscular utilization ratio (MUR), that compares the net joint moment obtained during gait to the maximal potential moment (MPM) at each percentage of the gait cycle, was used to estimate the mechanical relative demand. The MPM values were obtained by regression equations developed from torque data measured with a Biodex dynamometric system. The results showed that the peak MURs increased with gait cadence. The peak values were not significantly different between sides for all cadences despite mean absolute lateral differences ranging from 7% to 10%. The mean peak MURs of both sides ranged from 51.3% to 62.6%, from 20.7% to 49.9% and from 14.9% to 42.5%, for the plantarflexors, hip flexors and hip extensors, respectively. Highly significant associations were found between the MURs and net moments (numerator of the MUR ratio), with Pearson coefficients (r) superior to 0.80 for all muscles groups. The association between the MURs and the maximal potential moments (denominator of MUR ratio) was lower (0.010.05). CONCLUSION: The application of submaximal contractions of larger than 50% MVC is suitable to calibrate a simple EMG to force model for knee extension and flexion. This means that in clinical practice, the EMG to force model can be applied by patients who cannot exert maximal force. PMID- 15811614 TI - Fertility preservation: an emerging discipline in the care of young patients with cancer. PMID- 15811615 TI - Population-based cancer registries: the invisible key to cancer control. PMID- 15811616 TI - Fertility preservation for young patients with cancer: who is at risk and what can be offered? AB - Estimates suggest that by 2010, one in 715 people in the UK will have survived cancer during childhood. With increasing numbers of children cured, attention has focused on their quality of life. We discuss the causes of impaired fertility after cancer treatment in young people, and outline which patients are at risk and how their gonadal function should be assessed. With the report of a livebirth after orthotopic transplantation of cryopreserved ovarian tissue and the continued development of intracytoplasmic sperm injection for men with poor sperm quality, we assess established and experimental strategies to protect or restore fertility, and discuss the ethical and legal issues that arise. PMID- 15811617 TI - Cardiac tumours: diagnosis and management. AB - Primary cardiac tumours are rare, with an autopsy incidence ranging from 0.001% to 0.030%. Three-quarters of these tumours are benign and nearly half of the benign tumours are myxomas. Metastases to the heart are far more common than primary cardiac tumours. Primary cardiac tumours present with one or more of the symptoms of the classic triad of: cardiac symptoms and signs resulting from intracardiac obstruction; signs of systemic embolisation; and systemic or constitutional symptoms. They are diagnosed by use of transthoracic and transoesophageal echocardiograms, MRI, and CT scan. Whereas surgery is indicated in patients with benign tumours, systemic chemotherapy is indicated in those who have widespread or unresectable malignant disease, and chemotherapy and radiotherapy are usually combined in treatment of patients with primary cardiac lymphomas. The prognosis after surgery is usually excellent in the case of benign tumours but is unfortunately still limited in localised malignant diseases. Patients with sarcomas live for a mean of 3 months to 1 year, and those with lymphomas live up to 5 years if treated, but usually die within 1 month if untreated. PMID- 15811618 TI - Docetaxel for treatment of solid tumours: a systematic review of clinical data. AB - Docetaxel is a semisynthetic taxane, a class of anticancer agents that bind to beta tubulin, thereby stabilising microtubules and inducing cell-cycle arrest and apoptosis. Docetaxel was first approved for the treatment of anthracycline refractory metastatic breast cancer in the mid-1990s. Since then, several randomised trials have reported improved time-to-progression, overall survival, or both in metastatic breast cancer treated with single-agent docetaxel or docetaxel-based combination regimens. Data from two adjuvant trials have shown a survival benefit with the addition of docetaxel to standard anthracycline-based regimens in patients with high-risk early breast cancer. In four randomised studies, docetaxel improved survival in locally advanced or metastatic non-small cell lung cancer. Moreover, two trials have shown that docetaxel combined with estramustine or corticosteroids improves survival in metastatic androgen independent prostate cancer. Here, we review major randomised phase III trials with docetaxel in the treatment of solid malignant disease. PMID- 15811619 TI - Lessons learned from randomised clinical trials in adult low grade glioma. AB - "Lesson" is a Middle English word that has been defined as "a passage from sacred writings read in a service of worship" as well as "something learned by study or experience". The term is quite appropriate in assessment of what has been learned from randomised trials in adult low-grade gliomas, since the treatment of these tumours has traditionally been guided as much by belief as by fact. Therefore, when assessing these trials we can apply the principles of hermeneutics. Thus, the first meaning of "lesson" given here can be described as literal, whereas the second may be seen as figurative. Since hermeneutics may also refer to an in depth analysis of a particular text, the investigators will present their interpretation of data from randomised trials. The goal is to show that the lessons learned are not necessarily literal or dogmatic but can be much more allegorical in nature. PMID- 15811620 TI - Does the vagus nerve inform the brain about preclinical tumours and modulate them? AB - The inflammatory microenvironment is thought to play a pivotal part in tumorigenesis. But, can the brain be informed about peripheral preclinical cancer cells? Can it modulate tumour development? One of the key routes for information to reach the brain from visceral regions is through the vagus nerve. Yet, patients with ulcers who have had a vagotomy have been shown to die from cancer more frequently than do those who have not had this procedure, and surgical and chemical vagotomy attenuates tumour-induced anorexia and leads to enhanced tumour progression. We therefore postulate that the vagus nerve participates in informing the brain about tumorigenesis by transmiting information to the brain about tumour-associated proinflammatory cytokines. Furthermore, activation of the vagus could slow tumorigenesis by suppression of peripheral proinflammatory cytokines. PMID- 15811621 TI - Acquired resistance to imatinib in gastrointestinal stromal tumours caused by multiple KIT mutations. PMID- 15811622 TI - Tumour response to paclitaxel in an adult with relapsed nephroblastoma. PMID- 15811623 TI - Association of tissue-factor upregulation in squamous-cell carcinoma of the lung with increased tissue factor in circulating blood. PMID- 15811624 TI - Cloning and expression of hypoxia-inducible factor 1alpha from the hibernating ground squirrel, Spermophilus tridecemlineatus. AB - Mammalian hibernation is associated with apnoic breathing patterns and a hypoxia hypothermia connection has been suggested as part of the mechanism by which body temperature is reduced as animals enter torpor. Hence, we hypothesized that changes in the expression of the hypoxia inducible factor (HIF-1) may potentially be involved in regulating hibernation-responsive gene targets. The expression of the alpha subunit of HIF-1 was quantified at both gene and protein levels in four organs of the thirteen-lined ground squirrel, Spermophilus tridecemlineatus. Reverse transcription-PCR showed no change in hif-1alpha transcript levels in the liver, lung, skeletal muscle or brown adipose tissue of euthermic versus hibernating animals but HIF-1alpha protein levels were elevated by 60-70% in the two organs responsive for thermogenesis (brown adipose and skeletal muscle). Furthermore, assessment of DNA binding by HIF-1 in nuclear extracts from brown adipose revealed 6-fold higher levels in hibernator tissue than in euthermic controls suggesting increased expression of HIF-1 responsive genes during hibernation. The complete nucleotide sequence of hif-1alpha from ground squirrels, the first hif-1alpha sequence amplified from a hibernating mammal, was obtained using PCR amplification and 3' and 5' RACE. Amino acid sequence analysis revealed 90-95% identity with the HIF-1alpha protein from other mammals. Several unique amino acid sequence substitutions were identified that may affect protein conformation and could possibly function to counteract low temperature effects on HIF-1alpha conformation at near 0 degrees C body temperatures during torpor. PMID- 15811625 TI - Fission yeast mating-type switching: programmed damage and repair. AB - Mating-type switching in fission yeast follows similar rules as in budding yeast, but the underlying mechanisms are entirely different. Whilst the initiating double-strand cut in Saccharomyces cerevisiae requires recombinational repair for survival, the initial damage in Schizosaccharomyces pombe only affects a single strand, which can be sealed by gap repair in situ, whether or not it serves as an imprint for subsequent switching of mating type from an appropriate donor cassette. Recent papers have linked the transient stalling of a replication fork to the generation of a site-specific nick. This discontinuity then remains protected for a full cell cycle, until it interferes with replication in the next S-phase. It, thereby, represents a valuable model system to study the molecular safeguards to protect a replication fork at a predetermined hindrance to leading strand extension. The versatility of this experimental system has increased further yet by the recent development of a conditional setup, where imprinting and switching can be repressed or derepressed in response to external stimuli. PMID- 15811626 TI - DDB2, the xeroderma pigmentosum group E gene product, is directly ubiquitylated by Cullin 4A-based ubiquitin ligase complex. AB - Xeroderma pigmentosum (XP) is a genetic disease characterized by hypersensitivity to UV irradiation and high incidence of skin cancer caused by inherited defects in DNA repair. Mutational malfunction of damaged-DNA binding protein 2 (DDB2) causes the XP complementation group E (XP-E). DDB2 together with DDB1 comprises a heterodimer called DDB complex, which is involved in damaged-DNA binding and nucleotide excision repair. Interestingly, by screening for a cellular protein(s) that interacts with Cullin 4A (Cul4A), a key component of the ubiquitin ligase complex, we identified DDB1. Immunoprecipitation confirmed that Cul4A interacts with DDB1 and also associates with DDB2. To date, it has been reported that DDB2 is rapidly degraded after UV irradiation and that overproduction of Cul4A stimulates the ubiquitylation of DDB2 in the cells. However, as biochemical analysis using pure Cul4A-containing E3 is missing, it is still unknown whether the Cul4A complex directly ubiquitylates DDB2 or not. We thus purified the Cul4A containing E3 complex to near homogeneity and attempted to ubiquitylate DDB2 in vitro. The ubiquitylation of DDB2 was reconstituted using this pure E3 complex, indicating that DDB-Cul4A E3 complex in itself can ubiquitylate DDB2 directly. We also showed that an amino acid substitution, K244E, in DDB2 derived from a XP-E patient did not affect its ubiquitylation. PMID- 15811627 TI - New mammalian cellular systems to study mutations introduced at the break site by non-homologous end-joining. AB - The non-homologous end-joining (NHEJ) pathway is a mechanism to repair DNA double strand breaks, which can introduce mutations at repair sites. We constructed new cellular systems to specifically analyze sequence modifications occurring at the repair site. In particular, we looked for the presence of telomeric repeats at the repair junctions, since our previous work indicated that telomeric sequences could be inserted at break sites in germ-line cells during primate evolution. To induce specific DNA breaks, we used the I-SceI system of Saccharomyces cerevisiae or digestion with restriction enzymes. We isolated human and hamster cell lines containing the I-SceI target site integrated in a single chromosomal locus and we exposed the cells to a continuous expression of the I-SceI endonuclease gene. Additionally, we isolated human cell lines that expressed constitutively the I SceI endonuclease and we introduced the target site on an episomal plasmid stably transfected into the cells. These strategies allowed us to recover repair junctions in which the I-SceI target site was modified at high frequency (100% in hamster cells and about 70% in human cells). Finally, we analyzed junctions produced on an episomal plasmid linearized by restriction enzymes. In all the systems studied, sequence analysis of individual repair junctions showed that deletions were the most frequent modifications, being present in more than 80% of the junctions. On the episomal plasmids, the average deletion length was greater than at intrachromosomal sites. Insertions of nucleotides or deletions associated with insertions were rare events. Junction organization suggested different mechanisms of formation. To check for the insertion of telomeric sequences, we screened plasmid libraries representing about 3.5 x 10(5) junctions with a telomeric repeat probe. No positive clones were detected, suggesting that the addition of telomeric sequences during double strand break repair in somatic cells in culture is either a very rare event or does not occur at all. PMID- 15811628 TI - Artemis deficiency confers a DNA double-strand break repair defect and Artemis phosphorylation status is altered by DNA damage and cell cycle progression. AB - Mutations in the Artemis gene are causative in a subset of human severe combined immunodeficiencies (SCIDs) and Artemis-deficient cells exhibit radiation sensitivity and defective V(D)J recombination, implicating Artemis function in non-homologous end joining (NHEJ). Here we show that Artemis-deficient cells from Athabascan-speaking Native American SCID patients (SCIDA) display significantly elevated sensitivity to ionizing radiation (IR) but only a very subtle defect in DNA double-strand (DSB) break repair in contrast to the severe DSB repair defect of NHEJ-deficient cells. Primary human SCIDA fibroblasts accumulate and exhibit persistent arrest at both the G1/S and G2/M boundaries in response to IR, consistent with the presence of persistent DNA damage. Artemis protein is phosphorylated in a PI3-like kinase-dependent manner after either IR or a number of other DNA damaging treatments including etoposide, but SCIDA cells are not hypersensitive to treatment with etoposide. Inhibitor studies with various DNA damaging agents establish multiple phosphorylation states and suggest multiple kinases function in Artemis phosphorylation. We observe that Artemis phosphorylation occurs rapidly after irradiation like that of histone H2AX. However, unlike H2AX, Artemis de-phosphorylation is uncoupled from overall DNA repair and correlates instead with cell cycle progression to or through mitosis. Our results implicate a direct and non-redundant function of Artemis in the repair of a small subset of DNA double-strand breaks, possibly those with hairpin termini, which may account for the pronounced radiation sensitivity observed in Artemis-deficient cells. PMID- 15811629 TI - The UV-damaged DNA binding protein mediates efficient targeting of the nucleotide excision repair complex to UV-induced photo lesions. AB - Previous studies point to the XPC-hHR23B complex as the principal initiator of global genome nucleotide excision repair (NER) pathway, responsible for the repair of UV-induced cyclobutane pyrimidine dimers (CPD) and 6-4 photoproducts (6 4PP) in human cells. However, the UV-damaged DNA binding protein (UV-DDB) has also been proposed as a damage recognition factor involved in repair of UV photoproducts, especially CPD. Here, we show in human XP-E cells (UV-DDB deficient) that the incision complex formation at UV-induced lesions was severely diminished in locally damaged nuclear spots. Repair kinetics of CPD and 6-4PP in locally and globally UV-irradiated normal human and XP-E cells demonstrate that UV-DDB can mediate efficient targeting of XPC-hHR23B and other NER factors to 6 4PP. The data is consistent with a mechanism in which UV-DDB forms a stable complex when bound to a 6-4PP, allowing subsequent repair proteins--starting with XPC-hHR23B--to accumulate, and verify the lesion, resulting in efficient 6-4PP repair. These findings suggest that (i) UV-DDB accelerates repair of 6-4PP, and at later time points also CPD, (ii) the fraction of 6-4PP that can be bound by UV DDB is limited due to its low cellular quantity and fast UV dependent degradation, and (iii) in the absence of UV-DDB a slow XPC-hHR23B dependent pathway is capable to repair 6-4PP, and to some extent also CPD. PMID- 15811630 TI - The overexpression of specialized DNA polymerases in cancer. AB - Specialized DNA polymerases are required to bypass DNA damage lesions that would otherwise cause replication arrest and cell death. When operating on non canonical templates, such as undamaged DNA or on non-cognate lesions, these polymerases exhibit considerably reduced fidelity, resulting in the generation of mutations. Ectopic overexpression of these polymerases can also lead to an increased mutation rate and an enhanced capability of DNA repair, suggesting that they could potentially act as oncogenes if they were overexpressed in cancers. Here, we examine expression patterns of DNA polymerases in matched normal and tumor samples from a diverse range of tissues. As well as investigating the specialized polymerases beta, lambda, iota and kappa, we also investigate the expression of the replicative polymerases alpha, delta and epsilon. The data presented provide evidence for the overexpression of specialized polymerases in tumors, with more than 45% of the 68 tumor samples studied demonstrating greater than two-fold enhanced expression of at least one specialized polymerase. Of particular note, DNA polymerase beta (pol beta) was found to be overexpressed at both the mRNA and protein level in approximately one third of all tumor types studied, with overexpression being particularly frequent in uterus, ovary, prostate and stomach samples. Pols lambda, and iota were also found to be overexpressed to a significant extent in a range of tumor types, albeit less frequently than pol beta. In contrast, pol kappa was rarely found to be overexpressed in tumors but was found to be commonly underexpressed in many samples. Downregulation of pol beta expression by siRNA resulted in an increased sensitivity to the chemotherapeutic agent cisplatin, suggesting a role for this polymerase in providing tolerance to cisplatin-induced damage. These observations suggest that specialised DNA polymerases, and particularly pol beta, could be considered both as caretaker genes altered during tumorigenesis, and as potential drug targets to sensitise tumors to chemotherapy. PMID- 15811631 TI - Interstrand crosslink-induced homologous recombination carries an increased risk of deletions and insertions. AB - Homology directed repair (HDR) defends cells against the toxic effects of two ended double strand breaks (DSBs) and one-ended DSBs that arise when replication progression is inhibited, for example by encounter with DNA lesions such as interstrand crosslinks (ICLs). HDR can occur via various mechanisms, some of which are associated with an increased risk of concurrent sequence rearrangements that can lead to deletions, insertions, translocations and loss of heterozygosity. Here, we compared the risk of HDR-associated sequence rearrangements that occur spontaneously versus in response to exposure to an agent that induces ICLs. We describe the creation of two fluorescence-based direct repeat recombination substrates that have been targeted to the ROSA26 locus of embryonic stem cells, and that detect the major pathways of homologous recombination events, e.g., gene conversions with or without crossing over, repair of broken replication forks, and single strand annealing (SSA). SSA can be distinguished from other pathways by application of a matched pair of site specifically integrated substrates, one of which allows detection of SSA, and one that does not. We show that SSA is responsible for a significant proportion of spontaneous homologous recombination events at these substrates, suggesting that two-ended DSBs are a common spontaneous recombinogenic lesion. Interestingly, exposure to mitomycin C (an agent that induces ICLs) increases the proportion of HDR events associated with deletions and insertions. Given that many chemotherapeutics induce ICLs, these results have important implications in terms of the risk of chemotherapy-induced deleterious sequence rearrangements that could potentially contribute to secondary tumors. PMID- 15811632 TI - Suppression of gross chromosomal rearrangements by the multiple functions of the Mre11-Rad50-Xrs2 complex in Saccharomyces cerevisiae. AB - The Mre11-Rad50-Xrs2 complex in Saccharomyces cerevisiae has roles in the intra-S checkpoint, homologous recombination, non-homologous end joining, meiotic recombination, telomere maintenance and the suppression of gross chromosomal rearrangements (GCRs). The discovery of mutations in the genes encoding the human homologues of two MRX subunits that underlie the chromosome fragility syndromes, Ataxia telangiectasia-like disorder and Nijmegen breakage syndrome suggest that the MRX complex also functions in suppression of GCRs in human cells. Previously, we demonstrated that the deletion mutations in each of the MRX genes increased the rate of GCRs up to 1000-fold compared to wild-type rates. However, it has not been clear which molecular function of the MRX complex is important for suppression of GCRs. Here, we present evidence that at least three different activities of the MRX complex are important for suppression of GCRs. These include the nuclease activity of Mre11, an activity related to MRX complex formation and another activity that has a close link with the telomere maintenance function of the MRX complex. An activity related to MRX complex formation is especially important for the suppression of translocation type of GCRs. However, the non-homologous end joining function of MRX complex does not appear to participate in the suppression of GCRs. PMID- 15811633 TI - Error-prone DNA repair and translesion synthesis: focus on the replication fork. AB - Dean Rupp and Paul Howard-Flanders showed that, following exposure to ultraviolet light, bacteria deficient in nucleotide excision repair synthesised DNA with minimal delay and in pieces roughly the size of the distances between pyrimidine dimmers. The discontinuities or gaps between these pieces were subsequently sealed. This led directly to the hypothesis of translesion synthesis. PMID- 15811634 TI - Mending human genes: a job for a lifetime. AB - I have described a number of milestones along a career of nearly 40 years in DNA repair. Most important was the discovery that the human disease xeroderma pigmentosum represented mutations in various components of nucleotide excision repair. This ushered in a new field of research involving numerous investigators and which continues to expand and amaze. PMID- 15811635 TI - Modulation of cytokine production by Porphyromonas gingivalis in a macrophage and epithelial cell co-culture model. AB - Epithelial cells and macrophages play a major role in the host response to Porphyromonas gingivalis, a major etiologic agent of chronic periodontitis. Secretion of high levels of cytokines by these cells is believed to contribute to periodontal tissue destruction. To investigate the interactions between P. gingivalis and these two major cell types, we characterized the production of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and regulated on activation normal T cell expressed and secreted (RANTES) by an in vitro co-culture model composed of epithelial-like transformed cells (HeLa cell line) and macrophage-like cells (phorbol myristic acid-differentiated U937 cell line) following a challenge with different strains of P. gingivalis. P. gingivalis cells stimulated the secretion of pro-inflammatory cytokines (IL-1beta and IL-6) and chemokines (IL-8 and RANTES) in the co-culture model. Responses to P. gingivalis infection were influenced by the macrophage/epithelial cell ratios of the cultures. In addition, the level of secretion of these inflammatory mediators was dependent on the bacterial strain and the multiplicity of infection (MOI) used. The use of a gingipain-deficient mutant of P. gingivalis or the addition of a cysteine protease inhibitor suggested that the level of cytokines secreted by the co culture model was underestimated due to an extensive proteolytic degradation. This study showed that P. gingivalis can modulate the levels of inflammatory mediators, which may contribute to the progression of periodontitis. PMID- 15811636 TI - TGF-beta1 as a marker of delayed fracture healing. AB - Mitogens of the TGF-beta superfamily have been shown to be crucial local and systemic regulatory molecules involved in fracture healing. However, there exists only little information about systemic regulation of bone regeneration by growth factors and no reports comparing serum levels of bone growth factors between normal and failed fracture healing have been published so far. We hypothesized that quality of fracture healing might be reflected by systemic alterations of key regulatory growth factors involved in bone formation and remodeling. Therefore, the aim of this study was to evaluate possible differences in serum levels of BMP-2, BMP-4, and TGF-beta1 in patients with normal and delayed fracture healing. 103 patients with diaphyseal fractures of long bones were recruited prospectively. Peripheral blood samples were collected over a period of 6 months following a standardized time schedule. At the end of the individual investigation period, growth factor serum levels were measured using commercially available enzyme immunoassays. For the elimination of disturbing influences, patients in both groups were matched by gender, age, fracture type, and localization as well as applied technique of osteosynthesis. During a study period of 1 year, 10 patients with an atrophic type of delayed union could be retrieved and matched to 10 patients with normal fracture healing. The diagnosis of delayed union was assumed in case of failed consolidation 4 months after trauma. We found an increase of TGF-beta1 serum levels up to 2 weeks after fracture in both groups with a following return to the reference value within 6 weeks after trauma. However, decline of serum concentration occurred earlier in patients with delayed fracture healing. At 4 weeks after trauma, serum levels of TGF-beta1 were significantly lower in patients of the delayed union group. Serum levels of BMP-2 and BMP-4 were below detection level in all patients, respectively. These findings support the critical role of TGF-beta1 in fracture healing. Events during the consolidation phase seem to be dependent on sufficient availability of TGF-beta1. PMID- 15811638 TI - Unexpected problem of the traditional approach to the addition of aluminium hydroxide to vaccine preparations. PMID- 15811637 TI - Muscle atrophy and bone loss after 90 days' bed rest and the effects of flywheel resistive exercise and pamidronate: results from the LTBR study. AB - Muscle atrophy and bone loss pose substantial problems for long-term space flight and in clinical immobilization. We therefore tested the efficacy of flywheel resistive exercise and pamidronate to counteract such losses. Twenty five young healthy males underwent strict bed rest with -6 degrees head-down tilt for 90 days. Subjects were randomized into an exercise group that practiced resistive exercise with a 'flywheel' (FW) device every 2-3 days, a pamidronate group (Pam) that received 60 mg pamidronate i.v. 14 days prior to bed rest and a control group (Ctrl) that received none of these countermeasures. During the study, Ca(++) and protein intake were controlled. Peripheral quantitative computed tomography (pQCT) was used to assess bone mineral content (BMC) and muscle cross sectional area (mCSA) of calf and forearm. Measurements were taken twice during baseline data collection, after 28 and after 89 days bed rest, and after 14 days recovery. On the same days, urinary Pyridinoline excretion and serum levels of alkaline phosphatase, Ca(++) and PTH were measured. Pre-study exercise habits were assessed through the Freiburg questionnaire. Losses in calf mCSA were significantly reduced in FW (Ctrl: -25.6% +/- 2.5% Pam: -25.6% +/- 3.7%, FW: 17.3% +/- 2.7%), but not in the forearm mCSA (Ctrl: -6.4% +/- 4.33%, Pam: -7.7% +/- 4.1%, FW: -7.6% +/- 3.3%). Both diaphyseal and epiphyseal BMC losses of the tibia were mitigated in Pam and FW as compared to Ctrl, although this was significant only at the diaphysis. Inter-individual variability was significantly greater for changes in BMC than in mCSA, and correlation of BMC losses was poor among different locations of the tibia. A significant positive correlation was found between change in tibia epiphyseal BMC and serum cortisol levels. These findings suggest that both countermeasures are only partly effective to preserve BMC (FW and Pam) and mCSA (FW) of the lower leg during bed rest. The partial efficacy of flywheel exercise as well as the bones' response to unloading per se underlines the importance of mechanical stimuli. The huge variability of BMC changes, however, suggests that other factors affect changes in whole-bone strength following acute mechanical disuse. PMID- 15811639 TI - Comparison of antigen-specific cellular immune responses between individual PBMCs and splenocytes. PMID- 15811640 TI - Recombination and flavivirus vaccines: a commentary. PMID- 15811641 TI - Species-specificity of a murine immunocontraceptive utilising murine cytomegalovirus as a gene delivery vector. AB - Cytomegaloviruses are species-specific DNA viruses. Recombinant murine cytomegaloviruse (MCMV) expressing the mouse egg-coat protein zona pellucida 3 (mZP3) has been shown to sterilise female mice by breaking self-tolerance and inducing an immune response against the host ZP3. This virus has the potential to be used for mouse population control, however the effect of this recombinant immunocontraceptive virus in non-host species must be determined. Recombinant MCMV-mZP3, based on both laboratory and wild strains of virus, induced long-lived antibody responses against structural viral proteins and mZP3 when inoculated into laboratory rats, although no viral DNA or replicating virus was identified. The anti-mZP3 antibodies were specific for mouse ZP3, did not cross-react with rat ZP3, and had no effect on the fertility of the rats. PMID- 15811642 TI - The real cost of rabies post-exposure treatments. AB - The total costs to all payers, i.e., a societal perspective, of four rabies post exposure regimens were evaluated in two dog bite centres and four local health centres in India. Results showed that the Thai Red Cross intra-dermal regimen (TRC-ID), which uses only one-fifth of the IM dose of purified vero cell vaccine (PVRV) was at most 20% more expensive than use of Purified Chick Embryo Cell (PCEC) vaccine at one-tenth of the IM dose: this cost difference needs to be balanced with the small margin of safety of low potency doses. In local health centres where the staffs are not specially trained in rabies vaccination, the Zagreb intra-muscular regimen is an economical option. PMID- 15811643 TI - Induction of a Th2 immune response by co-administration of recombinant adenovirus vectors encoding amyloid beta-protein and GM-CSF. AB - Lines of experimental evidence indicate that induction of humoral immune responses in transgenic mouse models of Alzheimer disease (AD) by repeated injection of synthetic amyloid beta-protein (Abeta) is effective in prevention and clearance of deposits of Abeta aggregates in the brain of the mice. We have tested a non-injection modality whereby replication-defective adenovirus vectors encoding Abeta or the 99-amino acid carboxyl terminal fragment of Abeta precursor were intranasally administered to mice to elicit immune responses against Abeta. When mice were immunized only with the adenovirus vectors, immune responses against Abeta were negligible. By co-immunization with an adenovirus vector encoding granulocyte-macrophage colony stimulating factor (GM-CSF), the adenovirus vector encoding Abeta effectively elicited an immune response against Abeta. Immunoglobulin isotyping demonstrated a predominant IgG1 and IgG2b response, suggesting a Th2 anti-inflammatory type. Thus, adjuvantation is essential for induction of an immune response against Abeta by adenovirus mediated nasal vaccination. PMID- 15811644 TI - Fasciola hepatica procathepsin L3 protein expressed by a baculovirus recombinant can partly protect rats against fasciolosis. AB - Fasciola hepatica juveniles express immunodominant cathepsin L proteins, which are mainly found in their immature, procathepsin form. A gene encoding such a procathepsin L (FheCL3) was expressed by a baculovirus recombinant and by Saccharomyces cerevisiae. The glycosylated FheCL3 proteins obtained by both systems were used in a vaccination/challenge experiment in rats. Both antigens evoked similar antibody responses, but only the baculovirus expressed FheCL3 caused a significant protection against the number of liver flukes (52% protection, P=0.01), whereas the S. cerevisiae expressed FheCL3 did not. In a second experiment in rats, deglycosylated versions of both antigens were used, but this did not improve their efficacies. PMID- 15811646 TI - Interleukin-2 potentiates foot-and-mouth disease vaccinal immune responses in mice. AB - The present study describes the role of recombinant human interleukin-2 (rh IL-2) as immunomodulatory molecule in foot-and-mouth disease (FMD) vaccinal immune response in a murine model. The humoral immune response was evaluated by examining the antibody titre against FMD virus type O, A(22) and Asia 1 in serum samples obtained from different groups of mice inoculated with PBS, FMD vaccine alone; vaccine along with rh IL-2 on 0, 7, 14, 21, and 30 days post vaccination (DPV) by indirect double antibody Sandwich ELISA. The cellular immune response was also examined on different DPV by an MTT based lymphoproliferation assay in splenic mononuclear cells (SMNC) obtained from different groups. IL-2 was able to enhance the specific immune response against FMD virus type O, A(22) and Asia 1 as evident by significantly higher ELISA antibody titres (P<0.05) in serum obtained from mice receiving IL-2 along with vaccine as compared to mice immunized with vaccine alone. Similarly, the same group of mice showed significantly higher lymphoproliferative responses in SMNC against mitogen PHA and FMD virus types O, A(22) and Asia 1 on all DPVs as compared to the group inoculated with vaccine alone. PMID- 15811645 TI - Type I IFN is a powerful mucosal adjuvant for a selective intranasal vaccination against influenza virus in mice and affects antigen capture at mucosal level. AB - In view of the increasing interest in mucosal vaccination, we investigated whether type I IFN could act as adjuvant of an intranasally administered influenza vaccine. A single intranasal administration of IFNalphabeta-adjuvanted vaccine in anesthetized C3H/HeN mice was capable of protecting the totality of animals against virus challenge, while vaccine alone was only partially effective. To mimic intranasal vaccine administration in man and to limit vaccine delivery strictly to nasal mucosa, we used a second method of vaccination based on vaccine fractionation in six doses and intranasal instillation in non anesthetized mice. By using this vaccination schedule, IFNalphabeta-adjuvanted vaccine also prevented mice from disease development and induced an efficient long lasting immune response. Further experiments showed that IFNalphabeta increased the percentage of antigen-associated phagocytes in the nasal mucus layer, thus suggesting a new possible mechanism of action for type I IFN as an adjuvant. PMID- 15811647 TI - Changing attitudes towards polio vaccination: a randomized trial of an evidence based presentation versus a presentation from a polio survivor. AB - We compared the impact of epidemiological evidence and anecdotal evidence on changing vaccination attitudes amongst alternative medical students. Ninety-seven students were randomized to either an evidence-based lecture on the benefits of the polio vaccine on population health or a presentation from a visibly affected victim of polio. We compared change in responses to a survey measuring vaccination attitudes between the two groups. The follow-up rate was 73%. There was no statistically significant difference between the two groups in change in response to any of the survey questions. In a post hoc analysis we found that 25% of students were less likely to recommend the vaccine after being provided with evidence supporting vaccination. These findings suggest that confronting deeply held beliefs regarding vaccination may paradoxically strengthen these belief systems. PMID- 15811648 TI - Screening the whole genome of a pathogen in vivo for individual protective antigens. AB - We report the results of a general protocol that was used to screen the whole genome of Chlamydophila abortus, type strain B577 (formerly Chlamydia psittaci strain B577), in a mouse pneumonia model. Genetic immunization was used to functionally test the genes of C. abortus as vaccines in a mouse challenge system. Nine gene fragments were isolated that conferred protection, with five protecting as effectively as the live-vaccine positive control. Bioinformatics approaches were unable to reconstruct isolation of these antigens. These results suggest that pathogen genomes can be functionally screened for vaccine candidate antigens in a mouse model to reveal new classes of vaccine candidate antigens that may have therapeutic efficacy across host species, disease manifestations, and delivery platforms. PMID- 15811649 TI - Production of rabies neutralizing antibody in hen's eggs using a part of the G protein expressed in Escherichia coli. AB - In an attempt to produce anti-rabies immunoglobulin affordable for people living in developing countries, we have immunized layer chickens with a part of the G protein of rabies virus expressed in Escherichia coli. Immunoglobulin (IgY) was purified from the yolks of eggs layed by immunized hens. It was revealed in vitro that the antibody specifically bound to virions as well as cells infected with rabies virus. Moreover, the antibody apparently neutralized rabies virus infectivity. Inoculation of the antibody into mice infected with rabies virus reduced the mortality caused by the virus, suggesting that IgY directed to the part of the G protein expressed in E. coli could serve as a possible alternative to currently available anti-rabies human or equine immunoglobulins. PMID- 15811650 TI - Virus-like particles exhibit potential as a pan-filovirus vaccine for both Ebola and Marburg viral infections. AB - A safe and effective pan-filovirus vaccine is highly desirable since the filoviruses Ebola virus (EBOV) and Marburg virus (MARV) cause highly lethal disease typified by unimpeded viral replication and severe hemorrhagic fever. Previously, we showed that expression of the homologous glycoprotein (GP) and matrix protein VP40 from a single filovirus, either EBOV or MARV, resulted in formation of wild-type virus-like particles (VLPs) in mammalian cells. When used as a vaccine, the wild-type VLPs protected from homologous filovirus challenge. The aim of this work was to generate a multi-agent vaccine that would simultaneously protect against multiple and diverse members of the Filoviridae family. Our initial approach was to construct hybrid VLPs containing heterologous viral proteins, of EBOV and MARV, and test the efficacy of the hybrid VLPs in a guinea pig model. Our data indicate that vaccination with GP was required and sufficient to protect against a homologous filovirus challenge, as heterologous wild-type VLPs or hybrid VLPs that did not contain the homologous GP failed to protect. Alternately, we vaccinated guinea pigs with a mixture of wild-type Ebola and Marburg VLPs. Vaccination with a single dose of the multivalent VLP vaccine elicited strong immune responses to both viruses and protected animals against EBOV and MARV challenge. This work provides a critical foundation towards the development of a pan-filovirus vaccine that is safe and effective for use in primates and humans. PMID- 15811651 TI - Role of the N-linked glycans of the prM and E envelope proteins in tick-borne encephalitis virus particle secretion. AB - The tick-borne encephalitis (TBE) virus has two membrane glycoproteins (prM and E), which each has one N-linked glycan. Constructs that express prM and E proteins of TBE virus have been shown to produce virus-like particles (VLPs), which have surface properties that are similar to those of infectious viruses. To reveal the function of glycosylation of the TBE virus prM and E proteins in the secretion of VLPs, we expressed glycosylation-mutated prM and E proteins and compared the secretion levels and biological properties of the VLPs. In the prM protein glycosylation-deficient mutant, the level of secreted E protein was reduced to 60% of the wild-type level. On the other hand, in the E or prM-E protein glycosylation-deficient mutant, the level of secreted E protein was reduced to 10% of the wild-type level. Furthermore, the mutant which was glycosylated at positions 66 and 154 in protein E, the level of secreted E protein was four-fold higher than that of the wild-type. However, in the mutant which was glycosylated at position 66 only, E protein secretion was reduced to only 10% of the wild-type level. These data suggest that the glycan associated with the N-linked glycosylation site at position 154 in protein E plays an important role in VLP secretion. PMID- 15811652 TI - Associations of daycare and school entry vaccination requirements with varicella immunization rates. AB - School and daycare entry requirements have been credited with increasing immunization rates among school-age children, but no prior study has assessed the nationwide effects of entry requirements while controlling for individual, family, and household characteristics. The 2002 National Immunization Survey (NIS) is a nationally representative annual survey that includes provider record verified immunization dates for 20,546 children aged 19-35 months without prior history of varicella. In weighted bivariate and multivariate logistic regression models, we examined the association of state entry mandate implementation with children's up-to-date (UTD) status for varicella vaccine, adjusted for sociodemographic characteristics of children, mothers, and household income and for children's UTD status for other recommended vaccines. In this national sample representative of 5.6 million children, 83.2% (95% CI: 82.3%-84.1%) were UTD for varicella vaccine. Between 1997 and 2002 inclusive, 33 states and the District of Columbia had implemented school and/or daycare entry immunization mandates for varicella. In bivariate analyses, 84.9% (83.9%-85.9%) of children in states with varicella entry mandates were UTD, compared to 76.8% (75.3%-78.4%) of children in states without such mandates. In multivariate analyses controlling for child and family characteristics, children living in states with varicella entry mandates remained significantly more likely to be UTD for varicella than children in states without mandates. These findings indicate that immunization entry requirements are associated with higher immunization rates among preschool-age children, and suggest that the effects of entry requirements are independent of other individual and household factors associated with childhood immunization. PMID- 15811653 TI - Immune responses to recombinants of the South African vaccine strain of lumpy skin disease virus generated by using thymidine kinase gene insertion. AB - The South African vaccine strain of lumpy skin disease virus (type SA-Neethling) is currently being developed as a vector for recombinant vaccines of economically important livestock diseases throughout Africa. In this study, the feasibility of using the viral thymidine kinase gene as the site of insertion was investigated and recombinant viruses were evaluated in animal trials. Two separate recombinants were generated and selected for homogeneity expressing either the structural glycoprotein gene of bovine ephemeral fever virus (BEFV) or the two structural glycoprotein genes of Rift Valley fever virus (RVFV). Both recombinants incorporate the enhanced green fluorescent protein (EGFP) as a visual marker and the Escherichia coli guanine phosphoribosyl transferase (gpt) gene for dominant positive selection. The LSDV-RVFV recombinant construct (rLSDV RVFV) protected mice against virulent RVFV challenge. In a small-scale BEFV challenge cattle trial the rLSDV-BEFV construct failed to fully protect the cattle against virulent challenge, although both a humoral and cellular BEFV specific immune response was elicited. PMID- 15811654 TI - Models of Phase 1 vaccine trials: optimization of trial design to minimize risks of multiple serious adverse events. AB - A mathematical model of Phase 1 vaccine trial design was used to investigate strategies for minimizing the number of serious adverse events (SAEs) that could be encountered in the first Phase 1 trials of new vaccine formulations. For a relatively standard dose escalation trial with three dose groups each with 10 subjects, an optimal balanced between risk of more than one serious adverse event and trial design is achieved by splitting each dose group into two subgroups of three and seven. Based on the modeling, for a two vaccination, dose-escalating Phase 1 trial, a design where all subjects receive the first vaccination before any subject receives a second vaccination generally carries a lower risk of multiple serious adverse events than other designs. PMID- 15811655 TI - A human phase 1 vaccine clinical trial of the Plasmodium falciparum malaria vaccine candidate apical membrane antigen 1 in Montanide ISA720 adjuvant. AB - A dose escalating, placebo-controlled phase 1 trial was conducted to test the safety and immunogenicity of a vaccine containing recombinant Plasmodium falciparum apical membrane antigen 1 (AMA1) formulated in Montanide ISA720. Three groups of volunteers were vaccinated intramuscularly with 5 microg, 20 microg or 80 microg of AMA1, respectively, in 0.5 mL of formulation at 0, 3 and 6 months. Anti-AMA1 antibody levels and T cell stimulation indices were measured before and after each vaccination. No vaccine-related serious adverse events were recorded. Most subjects generated a mild to moderate, transient local reaction after the first vaccination. Three subjects developed a local reaction approximately 10 days following vaccination. Six of the 29 subjects seroconverted. Only one of these developed a high antibody titre. However, the interpretation of this trial was compromised by a loss of potency of the formulated vaccine during the course of the study. PMID- 15811656 TI - Subunit vaccine based on the p67 major surface protein of Theileria parva sporozoites reduces severity of infection derived from field tick challenge. AB - Two recombinant vaccines against Theileriaparva, based on a near full-length version of the sporozoite surface antigen p67 (p67(635)), or an 80 amino acid C terminal section (p67C), were evaluated by exposure of immunized cattle to natural tick challenge in two sites at the Kenya Coast and one in Central Kenya. Vaccination reduced severe ECF by 47% at the coast and by 52% in central Kenya from an average incidence of 0.53+/-0.07 (S.E.) in 50 non-immunised controls to an average of 0.27+/-0.05 in 83 immunised animals. The reduction in severe East Coast fever was similar to that observed in laboratory experiments with p67(635) and p67C. The p67 coding sequence from thirteen T. parva field isolates including seven from vaccinated cattle that were not protected, was 100% identical to the gene on which the recombinant vaccine is based, suggesting a predominantly homologous p67 antigenic challenge. The same parasite isolates were however genetically heterogeneous at several loci other than p67. PMID- 15811657 TI - Effect of formaldehyde on Cu(II) removal from synthetic complexed solutions by solvent extraction. AB - The effect of formaldehyde (HCHO) on the extraction of Cu(II) from an equimolar EDTA (ethylenediaminetetraacetic acid, H4L) solution with Aliquat 336 in kerosene (a quaternary amine) was examined. Experiments were carried out at different initial concentrations of Cu(II) (1.57-5 mol/m3), solution pH (3.0-8.0), HCHO concentrations (0-3 vol.%), Aliquat 336 concentrations (80-400 mol/m3), and temperatures (15-35 degrees C). It was shown that the distribution ratios (D) of Cu(II), which exists in the form of complexed anions CuL2-, increased with increasing equilibrium pH (pHeq), but reached a plateau at pHeq>4 for the system without HCHO and at pHeq>4.5 for the system with 1 vol.% HCHO. The D values increased with increasing HCHO concentration, likely due to the reduction of Cu(II) to Cu(I) by HCHO in solution. A semi-empirical two-parameter model was proposed to describe the extraction equilibrium, in which the non-ideality in organic phases was considered. The thermodynamic parameters were also evaluated and discussed. PMID- 15811658 TI - Control of VOCs emissions by condenser pre-treatment in a semiconductor fab. AB - The performance of a modified design of local condensers to pre-treat a variety of volatile organic compounds (VOCs) emitted from the stripping process of a semiconductor fab was tested in this study. The reaction temperature of the condensers was controlled at around 10 degrees C, it is relatively higher than the traditional condenser reaction temperature. Both VOCs and water vapors were condensed and formed liquid films. This resulted in an enhancement of the VOCs removals, especially for VOCs of high boiling points or solubility. This can help to prevent the follow up zeolite concentrator from damage. The performance of the integrated system of condenser/zeolite concentrator could, therefore, remain highly efficient for a longer operation time. Its annualized cost would also be lower than installing the zeolite concentrator only. PMID- 15811659 TI - Treating chemical mechanical polishing (CMP) wastewater by electro-coagulation flotation process with surfactant. AB - The effect of surfactants on the treatment of chemical mechanical polishing (CMP) wastewater by electro-coagulation-flotation (ECF) process was studied. Two surfactants, cetyltrimethylammonium bromide (CTAB) and sodium dodecylsulfate (SDS) were employed in this study to compare the effect of cationic (CTAB) and anodic (SDS) surfactants on ECF. The cationic surfactant can enhance the removal of the turbidity, but anodic surfactant cannot. It can be explained by the hetero coagulation theory. Moreover, the addition of CTAB in CMP wastewater can reduce the sludge volume and the flotation/sedimentation time in ECF process. The residual turbidity and dissolved silicon dropped with the increase of charge loading. No CTAB pollution problem exists after the ECF process. PMID- 15811660 TI - Prediction of the self-accelerating decomposition temperature (SADT) for liquid organic peroxides from differential scanning calorimetry (DSC) measurements. AB - We present a prediction (estimation, calculation, screening) method for the estimation of the self-accelerating decomposition temperature (SADT) for liquid organic peroxides from differential scanning calorimetry (DSC) measurements based on the concepts of thermal explosion theory originally introduced by Semonov which are adopted to our problem assuming nth-order reaction kinetics. For the peroxides under investigation, we demonstrate good agreement with the experimental SADT. This method can be used as a quick and easy applicable method for the estimation of the critical temperatures. PMID- 15811661 TI - Energetic co-ordination compounds: synthesis, characterization and thermolysis studies on bis-(5-nitro-2H-tetrazolato-N2)tetraammine cobalt(III) perchlorate (BNCP) and its new transition metal (Ni/Cu/Zn) perchlorate analogues. AB - Bis-(5-nitro-2H-tetrazolato-N2)tetraammine[cobalt(III)/nickel(III)] perchlorates (BNCP/BNNP) and mono-(5-nitro-H-tetrazolato-N)triammine [copper(II)/zinc(II)] perchlorates (MNCuP/MNZnP) have been synthesized during this work. The synthesis was carried out by addition of carbonato tetraammine metal [Co/Ni/Cu/Zn] nitrate [CTCN/CTNN/CTCuN/CTZnN] to the aqueous solution of sodium salt of 5 nitrotetrazole followed by reaction with perchloric acid. The precursors were synthesized by the reaction of aqueous solution of their respective nitrates with ammonium carbonate at 70 degrees C. The complexes and their precursors were characterized by determining metal and perchlorate content as well as infrared (IR), electron spectra for chemical analysis (ESCA) and X-ray diffraction (XRD) techniques. The TG profiles indicated that BNCP, BNNP and MNCuP are thermally stable up to the temperature of 260-278 degrees C unlike MNZnP (150 degrees C). Sudden exothermic decomposition was observed in case of bis-(5-nitro-2H tetrazolato-N2)tetraammine cobalt(III) perchlorate, bis-(5-nitro-2H-tetrazolato N2)tetraammine nickel(III) perchlorate and mono-(5-nitro-H-tetrazolato N)triammine zinc(II) perchlorate resulting in the severe damage of the sample cup. Sensitivity data indicated that the Co/Ni/Cu complexes are more friction sensitive (3-4.8 kg) than mono-(5-nitro-H-tetrazolato-N)triammine zinc(II) perchlorate (14 kg). The impact sensitivity results of the complexes corresponded to h50% of 30-36 cm. PMID- 15811662 TI - Solubility and stability of barium arsenate and barium hydrogen arsenate at 25 degrees C. AB - The inconsistency among current thermodynamic data of Ba3(AsO4)2(c) and BaHAsO4.H2O(c) led the authors to obtain independent solubility data of barium arsenate by both precipitation and dissolution experiments. Low and neutral pH (3.63-7.43) favored the formation of BaHAsO4.H2O(c). Both BaHAsO4.H2Oc and Ba3(AsO4)2(c) formed at the neutral pH conditions (7.47, 7.66), whereas Ba3(AsO4)2(c) was the only solid phase precipitated at high pH (13.03, 13.10). The Ba3(AsO4)2(c) precipitate acquired at 50 degrees C appeared as small leafy crystal, while the Ba3(AsO4)2(c) solid precipitated at 25 degrees C comprised granular aggregate with some smaller crystal clusters. XRD and SEM analyses of Ba3(AsO4)2(c) and BaHAsO4.H2O(c) indicated that the solids were indistinguishable before and after the dissolution experiments. In the present work, the solubility products (Ksp) for Ba3(AsO4)2(c) and BaHAsO4.H2O(c) were determined to be 10( 23.53)(10(-23.01) to 10(-24.00)) and 10(-5.60)(10(-5.23) to 10(-5.89)), respectively. DeltaGf degrees for Ba3(AsO4)2(c) and BaHAsO4.H2O(c) were calculated to be -3113.40 and -1544.47 kJ/mol, respectively. There was no difference between the solubility products of the leafy and the granular Ba3(AsO4)2(c) solids. PMID- 15811663 TI - The influence of the solvent on the epoxidation of cis,trans,trans-1,5,9 cyclododecatriene to trans-1,2-epoxy-cis,trans-5,9-cyclododecadiene by catalytic system tert-butyl hydroperoxide/Mo(CO)6. AB - The epoxidation of cis,trans,trans-1,5,9-cyclododecatriene to trans-1,2-epoxy- cis,trans-5,9-cyclododecadiene with the use of commercial solution of tert-butyl hydroperoxide in various organic solvents: tert-butyl peroxide, isooctane, decane, and nonane was investigated. The process was investigated in different experimental conditions. The best results were achieved with the application of tert-butyl hydroperoxide in isooctane. A significant influence of the nature of the solvent on the results of trans-1,2-epoxy-cis,trans-5,9-cyclododecadiene synthesis was found. PMID- 15811664 TI - Experimental determination of the minimum onset temperature of runaway reaction from a radioactive salt disposal in asphalt. AB - In order to clarify the reason for the most hazardous explosion in the history of the Japanese nuclear power development by a radioactive salt disposal in asphalt, an adiabatic process was developed using a Dewar vessel to minimize the temperature difference between the reactants and the surroundings. By this means, the heat evolution from a reaction which is readily lost can be detected at a lower temperature imitating the accidental condition. A series of ambient temperature-tracking Dewar experiments on asphalt salt mixtures were conducted under different initial ambient temperatures, such as 230, 210, 190, and 170 degrees C, respectively. As a result, it was observed that from 190 degrees C the sample's temperature rose until a runaway reaction occurred. The minimum onset temperature for the runaway reaction of the asphalt salt mixture was determined to be 190 degrees C, which is close to the initial temperature of approximately 180 degrees C, the same temperature as the real accident. This implies that at near this operational temperature, initial faint chemical reactions may occur and lead to further rapid reactions if heat is accumulated at this stage. PMID- 15811665 TI - Flammability limits and explosion characteristics of toluene-nitrous oxide mixtures. AB - Flammability limits and explosion characteristics of toluene-nitrous oxide mixtures are experimentally determined in an 8l spherical vessel, and are compared with corresponding values of toluene-air mixtures. The experiments, performed at atmospheric pressure and at an initial temperature of 70 degrees C, show that the flammable range of toluene in nitrous oxide (0.25-22.5 mol%) is about three times as wide as the corresponding range of toluene in air (1.3-7.1 mol%). Maximum values of the explosion pressure ratio and the deflagration index, K(G), are clearly higher when nitrous oxide is applied as an oxidizer. This can be attributed to the increased flame temperature and burning velocity of toluene nitrous oxide flames. Moreover, extremely high values of K(G) for near stoichiometric mixtures in combination with strong acoustic oscillations in the pressure signals of these mixtures indicate the existence of a flame accelerating mechanism. These phenomena are enhanced when an initial pressure of 6 bara is applied. Finally, when evaluating the lower flammability limit, it was found that pure nitrous oxide decomposes at pressures above 4.5 bara when applying an ignition energy of about 10 J. PMID- 15811666 TI - Endocrine disrupter--estradiol--in Chesapeake Bay tributaries. AB - Exogenous chemicals that interfere with natural hormonal functions are considered endocrine disrupting chemicals (EDCs). Estradiol (17beta-estradiol or E2) is the most potent of all xenoestrogens. Induction of vitellogenin (VTG) production in male fish occurs at E2 concentrations as low as 1 ng l-1. E2 reaches aquatic systems mainly through sewage and animal waste disposal. Surface water samples from ponds, rivers (Wicomico, Manokin and Pocomoke), sewage treatment plants (STPs), and coastal bays (Assawoman, Monie, Chincoteague, and Tangier Sound Chesapeake Bay) on the Eastern Shore of Maryland were analyzed for E2 using enzyme linked immuno-sorbent assay (ELISA). E2 concentrations in river waters varied between 1.9 and 6.0 ng l-1. Highest E2 concentrations in river waters were observed immediately downstream of STPs. E2 concentrations in all the coastal bays tested were 2.3-3.2 ng l-1. PMID- 15811667 TI - Combined use of photocatalyst and adsorbent for the removal of inorganic arsenic(III) and organoarsenic compounds from aqueous media. AB - A novel method for the removal of inorganic arsenic(III) (As(III)), monomethylarsonate (MMA), and dimethylarsinate (DMA) from aqueous media, was proposed and investigated. This method involves the combined use of TiO2 photocatalyst and an adsorbent, which has a high ability of As(V) adsorption, under photo-irradiation. When an aqueous solution of As(III) was stirred and irradiated by sunlight or xenon lamp in the presence of TiO2 suspension, the oxidation of As(III) into As(V) was effectively attained. By use of the same photocatalytic reaction, MMA and DMA were also degraded into As(V), while the total organic carbon (TOC) in the aqueous phase was decreased. When an aqueous solution of As(III) was stirred with a mixed suspension of TiO2 and an adsorbent for As(V) (activated alumina) under sunlight irradiation, the arsenic removal reached 89% after 24 h. By use of the same photocatalyst-adsorbent system, 98% of MMA and 97% of DMA were removed. The mechanism of the removal of arsenic species by the photocatalyst-adsorbent system was discussed. PMID- 15811668 TI - Removal of aniline from aqueous solution by PVC-CDAE ligand-exchanger. AB - The adsorption of aniline from aqueous solutions onto cobalt(II)-poly(vinyl chloride)-carboxylated diaminoethane (PVC-CDAE) resin has been studied using a mini-column apparatus at 25+/-0.1 degrees C. First of all, experimental data obtained from the breakthrough curves were tested by using the Scatchard plot analysis, to have a preliminary prediction about the types of interaction of the resin with aniline. Our aim was to determine the model which best describes the experimental data. The aspect of the Scatchard plot indicated that the aniline adsorption did not follow the Langmuir model and the presence of two types of binding sites for aniline on the resin. However, the dynamics of aniline uptake were represented by the Freundlich model reasonably well. The kinetics of aniline adsorption from aqueous solution on the cobalt(II)-PVC-CDAE have also been tested using continuous column runs and rate-controlling step of the process was determined. In this study, homogeneous diffusion model was adapted to a column system to describe the change in the aniline concentration at the column exit beginning from breakthrough point as a function of time. Kinetic studies revealed that the rate-controlling step of the aniline adsorption was predominately film diffusion controlled rather particle diffusion. PMID- 15811669 TI - Solid waste generation from oil and gas industries in United Arab Emirates. AB - Solid wastes generated from oil and gas industrial activities are very diverse in their characteristics, large in their amounts and many of which are hazardous in nature. Thus, quantifying and characterizing the generated amounts in association with their types, classes, sources, industrial activities, and their chemical and biological characteristics is an obvious mandate when evaluating the possible management practices. This paper discusses the types, amounts, generation units, and the factors related to solid waste generation from a major oil and gas field in the United Arab Emirates (Asab Field). The generated amounts are calculated based on a 1-year data collection survey and using a database software specially developed and customized for the current study. The average annual amount of total solid waste generated in the studied field is estimated at 4061 t. Such amount is found equivalent to 650 kg/capita, 0.37 kg/barrel oil, and 1.6 kg/m3 of extracted gas. The average annual amount of hazardous solid waste is estimated at 55 t and most of which (73%) is found to be generated from gas extraction-related activities. The majority of other industrial non-hazardous solid waste is generated from oil production-related activities (41%), The present analysis does also provide the estimated generation amounts per waste type and class, amounts of combustible, recyclable, and compostable wastes, and the amounts dumped in uncontrolled way as well as disposed into special hazardous landfill facilities. The results should help the decision makers in evaluating the best alternatives available to manage the solid wastes generated from the oil and gas industries. PMID- 15811670 TI - Evaluating the applicability of regulatory leaching tests for assessing the hazards of Pb-contaminated soils. AB - Soil contamination is a major environmental problem due to the ecological threat it poses. In this work, electron probe microanalysis (EPMA), X-ray diffraction (XRD), and leaching studies were employed to explain the different leaching behaviors of non-stabilized and stabilized soils. The applicability of the leaching fluids used in the toxicity characteristic leaching procedure (TCLP) and Australian Standards, AS 4439.1-1997 for assessing the hazards of contaminated soils was investigated as was the leaching of lead from soil stabilized by cement and buffered phosphate techniques. The results showed Pb speciation in the soil highly influenced metal leaching. The synthetic leaching fluids were unable to provide a reliable estimation of Pb concentration in the municipal landfill leachate (ML) due to the absence of organic ligands capable of forming stable complexes with the lead. Water provided the closest representation of lead leaching from the non-stabilized and phosphate stabilized soils while sodium tetraborate buffer was found to be suitable for cement-stabilized soil in a non putrescible landfill leachate system. A comparison of stabilization methods revealed that the buffered phosphate technique was more suitable for stabilizing the lead in the soil relative to cement stabilization. PMID- 15811671 TI - Solvent extraction applied to the recovery of heavy metals from galvanic sludge. AB - In this study, a hydrometallurgical treatment involving the solvent extraction and recovery of some heavy metals from a sulphuric acid leach solution of galvanic sludge, using di-(2-ethylhexyl)-phosphoric acid (D2EHPA) and bis-(2,4,4 trimethylpentyl)-phosphinic acid (Cyanex 272), both diluted in kerosene, has been investigated. The preliminary tests revealed the necessity to remove other metal species than zinc and nickel, contained in the leach solution, and therefore, processes to cement copper and precipitate chromium were then applied to finally obtain a Zn and Ni pregnant solution prior to solvent extraction. For the experimental conditions studied, Cyanex 272 showed a good recovery of Zn after the stripping stage using H2SO4, but D2EHPA effectively promoted a higher Zn extraction than Cyanex 272 did. The dependence of the solvent extraction method on variables such as pH, contact time and concentration of extractant, as well as the effect of different concentrations of sulphuric acid on stripping, are discussed. The discussion also includes the previous conditions developed to separate the main interfering metallic species from the leach solution in order to improve the extraction and recovery of zinc by solvent extraction. The final objective has been to achieve a solution as pure as possible to recover nickel sulphate. PMID- 15811672 TI - Zeta potential of soils with surfactants and its relevance to electrokinetic remediation. AB - There are numerous studies on the application of electrokinetic decontamination technique to remediate heavy metal contaminated fine-grained soils. In recent studies, surfactants have been used to increase the efficiency of contaminant removal. However, there is limited data available on how physicochemical parameters such as zeta potential (zeta) of soils changes in the presence of surfactants. Understanding the zeta potential variations of soils with surfactant addition is important because it controls the direction and magnitude of electro osmotic permeability, which plays important role on the efficiency of electrokinetic remediation. In this study, zeta potentials of kaolinite, montmorillonite and quartz powder with Li+, Ca+2, Cu+2, Pb+2 and Al+3 in the presence of anionic, cationic and non-ionic surfactants were determined. The results indicate that anionic surfactants produce negative zeta potentials. The other surfactants produce both positive and negative zeta potentials depending on soil type and ion present in the system. The results also indicate that the zeta potential of kaolinite and quartz powder with surfactants showed similar trends; however, the absolute magnitude of the zeta potential of quartz powder is higher than that of kaolinite. The zeta potential of montmorillonite commonly shows a different trend from those of kaolinite and quartz powder. Based on the test results, it is recommended that zeta potential of soils be determined before the electrokinetic decontamination in order to maximize the efficiency of the technique. PMID- 15811673 TI - Biodegradation of total organic carbons (TOC) in Jordanian petroleum sludge. AB - Biodegradation is cost-effective, environmentally friendly treatment for oily contaminated sites by the use of microorganisms. In this study, laboratory experiments were conducted to establish the performance of bacterial isolates in degradation of organic compounds contained in oily sludge from the Jordanian Oil Refinery plant. As a result of the laboratory screening, three natural bacterial consortia capable of degrading total organic carbons (TOC) were prepared from isolates enriched from the oil sludge. Experiments were conducted in Erlenmeyer flasks under aerobic conditions, with TOC removal percentage varied from 0.3 to 28% depending on consortia type and concentration. Consortia 7B and 13B exhibited the highest TOC removal percentage of 28 and 22%, respectively, before nutrient addition. TOC removal rate was enhanced after addition of nutrients to incubated flasks. The highest TOC reduction (43%) was estimated after addition of combination of nitrogen, phosphorus and sulphur to consortia 7B. A significant variation (P<0.005) was observed between the effect of consortia type and concentration on TOC% reduction. No significant variation was observed between incubation at 10 and 18 days in TOC% reduction. This is the first report concerning biological treatment of TOC by bacteria isolated from the oil refinery plants, where it lays the ground for full integrated studies recommended for the degradation of organic compounds that assist in solving sludge problems. PMID- 15811674 TI - Preparation of TiO2-SiO2 mixed gel spheres for strontium adsorption. AB - A simple external gelation process, taking full advantage of the gelation features of titanium and silica, was developed to prepare TiO2-SiO2 mixed gel spheres suitable for strontium adsorption. The source solutions used for the process were prepared from different mixtures of 1M TiCl4 and 1M Na2SiO3 solutions and converted into droplets in a gelation column. The suitable spheres for strontium adsorption were obtained using a hexone (methyl isobutyl ketone) solution as the drop formation medium and ammonia as the gelling agent. The mixed oxide gels were identified and characterized by DTA/TGA, FTIR and XRD analysis. The parameters affecting the strontium adsorption, such as weight ratio of TiO2, pH, temperature, shaking time and selectivity towards competing ions were investigated. Sorption data have been interpreted in terms of Freundlich, Langmuir and Dubinin-Radushkevich equations. Thermodynamic parameters for the sorption system have been determined at four different temperatures. The value of DeltaH degrees =39.553 kJ/mol and DeltaG degrees =-16.687 kJ/mol at 296 K prove that the sorption of strontium on mixed oxide gel is an endothermic and a spontaneous process. PMID- 15811675 TI - In situ ozonation of anthracene in unsaturated porous media. AB - Soil column experiments were conducted to investigate the effect of ozonation duration, contaminant content, particle size, moisture content, OH radical scavenger and soil organic matter on the removal of anthracene by in situ ozonation. In the whole study, the gas flow rate was 100 mL/min and concentration of gaseous ozone was 40 mg/L. The removal efficiency increased with the elapsed time, but the removal rate decreased in the range of 0-90 min. As anthracene content in sand decreased from 50 to 10 mg/kg, the removal efficiency increased from 42.1% to 62.0%, and ozone passed through soil column more rapidly. However, the ozone effectiveness reduced when anthracene content dropped. Small particle size provides a large interfacial area, which led to the high removal efficiency and long ozone breakthrough time in the column. The profile of residual anthracene in soil column varied more greatly at smaller particle size. The removal efficiency reduced when the moisture content rose from 0% to 9.1%. The ozone breakthrough time also decreased with the increasing moisture content. The presence of sodium bicarbonate or humic acid reduced the removal efficiency to some extent. GC-MS was employed in this study to determine 9,10-anthraquinone as the main ozonation product. PMID- 15811676 TI - Laboratory batch experiments of the combined effects of ultrasound and air stripping in removing CCl4 and 1,1,1-TCA from water. AB - Ultrasonic and air-stripping techniques for removal of carbon tetrachloride (CCl4) and 1,1,1-trichloroethane (1,1,1-TCA) from water were studied in batch experiments. Ultrasound (US) is effective for destroying organic compounds in aqueous solutions whereas air stripping (AS) efficiently transfers volatile compounds from the liquid to the gas phase. In simultaneous US and AS experiments, synergistic effects were observed and attributed to the effect of US on the mass transfer process. Using a photographic method, ultrasonic break up of gas bubbles and changes in gas holdup ratios were examined. In the two different gas-sparging systems studied, ultrasonic waves did not break up gas bubbles. In contrast, bubbles from the smaller porous size diffuser were coalesced due to sonication. In addition, both photographic and gas holdup experiments demonstrated that ultrasonic irradiation increased the gas holdup ratio. The enhancement observed in the removal of the compounds appeared to be due to this greater ultrasonic gas holdup ratio. PMID- 15811677 TI - Effects of calcium competition on lead sorption by palm kernel fibre. AB - The kinetics of sorption of a mono-solute of lead ions and of a bi-solute of lead and calcium ions onto palm kernel fibre was investigated in a batch system. The experimental data were analysed based on an intraparticle diffusion equation and a pseudo-second-order mechanism, in both the mono- and bi-solute sorption systems, in order to predict the rate constant of sorption, the equilibrium capacity, and the initial sorption rate. The results indicate that the sorption mechanism is described by a pseudo-second-order equation. Intraparticle diffusion was significant in the lower-concentration systems. In addition, a modified intraparticle diffusion equation was applied to the sorption systems. PMID- 15811678 TI - Sorption of an anionic dye by uncalcined and calcined layered double hydroxides: a case study. AB - Layered double hydroxides (LDHs) with a Mg/Al molar ratio of 2:1 were synthesized by using a co-precipitation method and their calcined products (CLDHs) were obtained by heating Mg/Al-LDHs at 500 degrees C. Sorption of a weak acid dye, Brilliant Blue R (BBR), by LDHs and CLDHs indicated equilibrium time required for BBR sorption by CLDHs was less than 12h, regardless of initial concentration of BBR, whereas BBR sorption by LDHs was longer than 20 h. Sorption capacity of CLDHs was much larger than that of LDHs. Therefore, CLDHs could be used to remove anionic dyes of relatively high concentrations while LDHs may only be used to remove anionic dyes of low concentrations. Isotherms for BBR sorption by CLDHs and LDHs were well described using the Freundlich and Langmuir equations, respectively. When the initial pH of BBR solutions was lower than 8.0, the final pH of the solution after sorption was enhanced and stabilized at 10.6-10.8. The effect of initial pH (<8.0) on BBR removal was negligible, which would be environmentally important for precipitation/co-precipitation of co-existing metal cations. The effects of both Cl- and SO4(2-) on BBR sorption by CLDHs were minimal; but the presence of CO(3)2- markedly reduced BBR removal. Thermal regeneration for re-use of LDHs and CLDHs after BBR sorption was feasible only within the first two cycles, after which the regenerated materials suffered from a large loss in their sorption capacities. PMID- 15811679 TI - Structural and ionization effects on the adsorption behaviors of some anilinic compounds from aqueous solution onto high-area carbon-cloth. AB - The adsorption of anilinic compounds; aniline, p-toluidine, 1-napthylamine and sodium salt of diphenylamine-4-sulfonic acid from solutions in H2O, in 1 M H2SO4 or in 0.1 M NaOH onto activated carbon-cloth was studied by in situ UV spectroscopy. A specially designed adsorption cell was used for this purpose. The adsorption processes were found to follow first-order kinetics and the rate constants were determined. The pH at the point of zero charge of the carbon-cloth surface was measured as 7.4. The highest rates and extents of adsorption were observed from H2O solutions for aniline, p-toluidine and 1-napthylamine and from solution in 1 M H2SO4 for the sodium salt of diphenylamine-4-sulfonic acid. The adsorption behaviors of these four anilinic compounds in the three solutions (in H2O, in 0.1 M NaOH and in 1 M H2SO4) were explained in terms of electrostatic and dispersion interactions between carbon-cloth surface and the anilinic species. The adsorption isotherm data for the anilinic compounds were derived at 30 degrees C and treated according to Langmuir and Freundlich models. The Freundlich model was found to represent the experimental isotherm data better than Langmuir model. PMID- 15811680 TI - Aqueous solubility and alkaline hydrolysis of the novel high explosive hexanitrohexaazaisowurtzitane (CL-20). AB - The recently developed polycyclic nitramine CL-20 is considered as a possible replacement for the monocyclic nitramines RDX and HMX. The present study reports aqueous solubility data for CL-20, as well as the kinetic parameters for its alkaline hydrolysis with sodium hydroxide below and above its solubility limits. Aqueous solubility of CL-20 was measured in the temperature range of 4-69 degrees C and the data were fitted to a generalized solubility model. Alkaline hydrolysis experiments were conducted at 15, 20, 30 and 40 degrees C, with hydroxide concentrations ranging from 0.25 to 300 mM. Like RDX and HMX, alkaline hydrolysis of CL-20 follows second-order kinetics. CL-20 alkaline hydrolysis was found to proceed at a significantly faster rate than RDX. The temperature dependency of the second-order rate constants was evaluated using the Arrhenius model. The activation energy for CL-20 was found to be within close range of the activation energies reported for RDX and HMX. PMID- 15811681 TI - Study of Cu(II) biosorption by dried activated sludge: effect of physico-chemical environment and kinetics study. AB - Biosorption is a recent technology used to remove heavy metal ions from aqueous solutions. The biosorption of copper ions from aqueous solution by dried activated sludge was investigated in batch systems. Effect of solution pH, initial metal concentration and particle size range were determined. The suitable pH and temperature for studied conditions were determined as 4.0 and 20 degrees C, respectively. The theoretical max biosorption capacity of activated sludge was 294 mg g-1 at 20 degrees C for <0.063 mm particle size. The equilibrium data fitted very well to both Langmuir and Freundlich isotherm models. The pseudo first and second-order kinetic models were used to describe the kinetic data. The experimental data fitted to second-order kinetic model. The particle size and initial metal concentration were effected the biosorption capacity of dried activated sludge. An increase in the initial metal concentration increases of biosorption capacity, which also increases with decreasing particle size. Dried activated sludge has different functional groups according to the FT-IR results. PMID- 15811682 TI - Electrolytic recovery of lead in used lime leachate from municipal waste incinerator. AB - The purification of municipal incinerator gases generates huge quantities of both fly ashes and used lime, containing high amount of heavy metals (Pb, Cd, Zn and Hg). The used lime is mainly contaminated by Pb and often considered as hazardous waste. This paper is related to a laboratory study on the performance of electrochemical techniques used to recover Pb from leachate produced during used lime decontamination. Two types of electrolytic units (monopolar and bipolar electrode cells) using different electrode materials were studied. Effectiveness was measured in terms of energy consumed, weight of residue sludge produced and reduction in Pb concentration. Results showed that the best performances for Pb removal (in terms of effectiveness and cost) were obtained by initially adjusting the pH of UL leachate to pH 7.5-8.5 with sulphuric acid followed by electrochemical treatment using bipolar electrode (mild steel) system operated at current intensity of 1.5A through only 5 min. The yield of Pb removal was 98.8+/ 1.3% and an amount of 4.9+/-1.3 kg trt-1 of metallic sludge was produced. The optimal conditions determined for Pb recovery involved a total cost of CAN$ 1.66 t-1 of dry residue treated, including acid consumption, energy consumption and metallic sludge disposal. PMID- 15811683 TI - The facile flow-injection spectrophotometric detection of gold(III) in water and pharmaceutical samples using 3,5-dimethoxy-4-hydroxy-2-aminoacetophenone isonicotinoyl hydrazone (3,5-DMHAAINH). AB - A simple, sensitive and rapid flow-injection spectrophotometric method was developed for the determination of trace amounts of Au(III) in aqueous dimethylformamide (DMF). The method is based on formation of Au(III)-(3,5 DMHAAINH)3 complex. The optimum conditions for the chromogenic reaction of Au(III) with 3,5-DMHAAINH is studied and the colored (reddish brown) complex is selectively monitored at lambda(max) 490 nm at pH 6.0. The reaction and flow conditions of the full experimental design were optimized. The detection limit (2 s) of 0.1 microg l-1 Au(III) was obtained at a sampling rate of 15 samples h-1. Beer's law is obeyed over the range of 0.30-4.00 microg ml-1. The molar absorptivity and Sandell's sensitivity were 3.450x10(4) M and 0.0050 microg ml-1, respectively. Job's method of continuous variation and stability constants corresponding to these maxima was determined and found to be 9.3x10(15) (1:3, M:R) (M, metal; R, reagent). The detailed study of various interferences confirmed the high selectivity of the developed method. The method was successfully applied for the determination of trace amount of Au(III) in water and pharmaceutical samples. The results obtained were in agreement with the reported methods at the 95% confidence level. PMID- 15811684 TI - Comparative photocatalytic study of two selected pesticide derivatives, indole-3 acetic acid and indole-3-butyric acid in aqueous suspensions of titanium dioxide. AB - Heterogeneous photocatalysed degradation of two selected pesticide derivatives such as indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) has been investigated in aqueous suspensions of titanium dioxide by monitoring the change in substrate concentration employing UV spectroscopic analysis technique and depletion in total organic carbon (TOC) content as a function of irradiation time. The degradation kinetics was studied under different conditions such as pH, types of TiO2) substrate and catalyst concentration, and in the presence of electron acceptor such as hydrogen peroxide (H2O2) besides molecular oxygen. The degradation rates were found to be strongly influenced by all the above parameters. The photocatalyst Degussa P25 showed comparatively highest photocatalytics. The pesticide derivative, indole-3-acetic acid was found to degrade slightly faster than indole-3-butyric acid. PMID- 15811685 TI - Characterization of precursors to trihalomethanes formation in Bangkok source water. AB - Resin adsorption techniques using three types of resin (DAX-8, AG-MP-50, and WA 10) were employed to characterize the raw water (RW) from the major 3 million m3/day (793 million gal/day) drinking water treatment plant in Bangkok, Thailand. The dissolved organic carbon (DOC) mass distribution sequences of the six organic fractions in raw water, from high to low, were hydrophilic neutral (HPIN), hydrophobic acid (HPOA), hydrophilic acid (HPIA), hydrophobic neutral (HPON), hydrophilic base (HPIB), and hydrophobic base (HPOB). HPIN and HPOA were the two main precursors for trihalomethanes formation (THMFP) in this water source following chlorination. The chlorination of HPON and HPIN fractions only led to the formation of mostly chloroform, while other organic fractions formed both chloroform and bromodichloromethane. The linear dependency between each organic fraction concentration and THMFP indicated that the reactions of each organic fraction with chlorine were first-order. PMID- 15811686 TI - Removal of organic compounds by alginate gel beads with entrapped activated carbon. AB - The adsorption of alginate gel (AG) beads and AG with activated carbon entrapped (AG-AC) beads prepared using different types of metal ions were investigated by measuring the removal of several organic compounds with different charges and size. AG-AC beads prepared in a CaCl2 solution adsorbed strongly positively charged compounds as well as electrically neutral and low molecular weight compounds such as p-chlorophenol. However, a high molecular weight humic acid was not adsorbed by AG-AC. The AG-AC selectively adsorbed p-chlorophenol from a humic acid solution. The adsorption capacity obtained from the adsorption isotherm of AC entrapped in AG was compared with that of AC. The AG-AC beads prepared in a solution of FeCl3 were able to specifically adsorb negatively charged gallic acid. Thus, entrapping AC into AG resulted in the selective adsorption. PMID- 15811687 TI - Adsorption and thermochemical data of divalent cations onto silica gel surface modified with humic acid at solid/liquid interface. AB - Humic acid immobilized onto silica gel surface was studied by the calorimetric titration of divalent cations in aqueous solution. The adsorption isotherms were obtained by the batchwise method and were fitted to a modified Langmuir equation. The maximum number of moles per gram of the material gave: 10.42+/-0.75, 13.16+/ 0.58, 7.87+/-0.58 for copper, nickel and zinc, respectively. Gibbs free energies were negative for all systems and the adsorption interactions calorimetrically followed presented endothermic enthalpic values: 6.24+/-0.47, 6.75+/-0.74, 6.97+/ 0.58 kJ mol-1 for the same sequence of divalent cations. All liquid/solid interface adsorptions were entropically driven. PMID- 15811688 TI - Removal of copper from industrial sludge by traditional and microwave acid extraction. AB - This work elucidates the removal of copper from industrial sludge by traditional and microwave acid extraction. The effects of acid concentration, extraction time, sludge particle size and solid/liquid (S/L) ratio on copper removal efficiency were evaluated. Leaching with more concentrated acid yielded greater copper content from the industrial sludge. The experimental findings reveal that the most economical traditional extraction conditions were the use of 1N sulfuric or nitric acid for 60 min at an S/L ratio of 1/20; however, at an S/L ratio of 1/6, the extraction time needed to achieve the same copper removal efficiency was increased to 36 h. Increasing the microwave power and reducing the S/L ratio increased the copper extraction efficiency and the effect in the larger S/L ratio system was more significant. A comparison of the results of microwave-assisted (microwave only) and microwave-enhanced (microwave with addition of active carbon) acid extraction demonstrated that under both conditions, S/L ratio=1/6 and 1/20; adding active carbon shortened the extraction time required to achieve 80% copper extraction efficiency from 20 to 10 min. These experimental results indicate that the most important factors that most strongly affected microwave acid extraction were the addition of a microwave absorber, the microwave power input and the S/L ratio. The sludge particle size did not significantly affect the copper extraction. The results reveal that sulfuric acid was an effective extractant and that the copper fraction in the extracted sludge shifted from being mostly bound to the Fe-Mn oxides and organic matter, to being mostly bound to organic matter and remaining as a residue during acid extraction. PMID- 15811689 TI - Photodegradation of tetra- and hexachlorodibenzo-p-dioxins. AB - This study examined the direct photolysis and photocatalytic processes for 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and 1,2,3,6,7,8 hexachlorodibenzo-p-dioxin (1,2,3,6,7,8-HxCDD). The photocatalytic procedure was performed on the TiO2 film under irradiation with 365 nm UV and the compounds were immobilized on TiO2/solid phase. No 2,3,7,8-substituted PCDD/Fs products were detected in photocatalytic process under the experimental conditions. The reaction rate constants were 0.3256 h-1 for 2,3,7,8-TCDD (2000 ng) in UV/TiO2 reaction, 0.2474 h-1 for 1,2,3,6,7,8-HxCDD (2000 ng) in UV/TiO2 reaction and 0.0666 h-1 for 1,2,3,6,7,8-HxCDD (50 ng) under direct UV irradiation. For 1,2,3,6,7,8-HxCDD (50 ng) in a UV/TiO2 reaction, the degradation is too fast to determine the reaction rate. The photocatalytic process was faster than direct photolysis for the same chlorinated PCDDs, and the rate decreased with increasing PCDDs quantity. The photocatalytic rate of the PCDDs decreased with increasing chlorination extent. The confirmed intermediates of 1,2,3,6,7,8-HxCDD in direct photolysis, 1,2,3,7,8-PeCDD and 2,3,7,8-TCDD both were formed by the loss of a longitudinal chlorine nearest the oxygen atom. The quantity of 1,2,3,7,8-PeCDD and toxic equivalency quantity (TEQ) declined after 10 h of UV irradiation. The proposed dechlorination pathway of 1,2,3,6,7,8-HxCDD was via 1,2,3,7,8-PeCDD to 2,3,7,8-TCDD. Formation of trace concentrations of 1,2,3,4,7,8-hexachlorodibenzo p-dioxin (1,2,3,4,7,8-HxCDD) and 1,2,3,7,8,9-hexachlorodibenzo-p-dioxin (1,2,3,7,8,9-HxCDD) from 1,2,3,6,7,8-HxCDD appears to be a minor side reaction. PMID- 15811690 TI - Characterization of alpha-cordierite glass-ceramics from fly ash. AB - Batches of alpha-cordierite glass-ceramics, designated as GC-I and GC-II, containing 68 and 64 wt.% fly ash, respectively, were crystallized in the temperature range of 1125-1320 degrees C. The XRD (X-ray powder diffractometer) of the glass-ceramics show that alpha-cordierite became the dominant phase in GC I and GC-II at 1200 degrees C. GC-I and GC-II, whose solid parts contain 74 and 78 vol.% alpha-cordierite and whose compressive strengths are 35 and 50 MPa, respectively, have the respective linear thermal expansion coefficients of 1.51x10(-6) and 1.43x10(-6)/ degrees C. The fly ash alpha-cordierite glass ceramics can be employed as kiln furniture, honeycomb substrates for catalysts, and heat exchangers. PMID- 15811691 TI - Prediction of simultaneous adsorption of Cu(II) and Pb(II) onto activated carbon by conventional Langmuir type equations. AB - Removal of Cu(II) and Pb(II) by adsorption onto activated carbon was examined in single- and binary-component aqueous solutions representative of contaminated solutions containing heavy metals. Reversibility of adsorption of the heavy metals on the activated carbon was evaluated by desorption experiments. The number of the maximum adsorption sites and adsorption equilibrium constants of Cu(II) and Pb(II) were estimated by the results of single-component systems assuming the Langmuir adsorption model. The adsorption sites per gram of activated carbon resulted in similar values for Cu(II) and Pb(II) from the isotherms. The adsorption constant for Pb(II) was nearly 1.8 times greater than that of Cu(II). Rate constants of adsorption and desorption were also estimated from the kinetic analysis. Using the single set of common parameters obtained from the single-component systems, the experimental results for a binary component system were quantitatively predicted. Competitive adsorption of Cu(II) and Pb(II) on the same adsorption sites was confirmed by both experimental and predicted results of adsorption in the binary mixture. PMID- 15811692 TI - Reply to comment on "Nitrate removal from aqueous solution by adsorption onto various materials", by Y.S. Ho. PMID- 15811693 TI - Acoustic reflexes to Schroeder-phase harmonic complexes in normal-hearing and hearing-impaired individuals. AB - Harmonic complexes generated with positive or negative Schroeder-phases may result in differences in cochlear excitation, even though their long-term spectra and amplitudes are equal. As a measure of possible differences in cochlear excitation resulting from these harmonic complexes, thresholds and growth of the acoustic reflex were assessed in normal-hearing and hearing-impaired subjects. Harmonic complexes with fundamental frequencies of 50, 100, and 200 Hz were constructed with positive and negative-Schroeder phases. In normal-hearing subjects, acoustic reflex thresholds for the 50- and 100-Hz fundamental waveforms were typically lower for negative Schroeder-phase complexes than for positive Schroeder phase stimuli. At the highest fundamental frequency of 200 Hz, there were no significant threshold differences due to phase. Hearing-impaired subjects showed a similar pattern for thresholds between the two phase selections, but with smaller differences than those observed in normal-hearing subjects. At levels above reflex threshold, the magnitude of the acoustic reflex was greater for the negative-phase than the positive-phase stimuli for the lowest fundamental frequency, but no significant differences were observed at fundamental frequencies of 100 and 200 Hz. These results are consistent with generally greater cochlear excitation in response to negative than to positive Schroeder phase stimuli when the fundamental frequency is sufficiently low. Increased excitation may reflect a synchronization of response across a wide band of frequencies in the cochlea when the rate of frequency sweep within periods of these harmonic complexes is appropriately matched to timing characteristics of the traveling wave. PMID- 15811694 TI - Vibration induced hearing loss in guinea pig cochlea: expression of TNF-alpha and VEGF. AB - Transcranial vibration was applied for seven animals at a frequency of 250 Hz for 15 min, and five animals were used as normal controls to investigate cellular and molecular mechanism linked to vibration-induced hearing loss in animal model. Compound action potential (CAP) thresholds were measured by round window niche electrode. The expression of tumour necrosis factor alpha (TNF-alpha) and its receptors (TNF R1, TNF R2), vascular endothelium growth factor (VEGF) and its receptors (VEGF R1, VEGF R2) were analysed by immunohistochemistry. Transcranial vibration caused expression of TNF-alpha, TNF R1 and TNF R2 in the cochlea and the expression of TNF R2 was stronger than that of TNF R1. Vibration also induced VEGF and VEGF R2 expression in the cochlea. The average immediate hearing loss was 62 dB and after three days still 48 dB. It is concluded that transcranial vibration as during temporal bone drilling produces cochlear shear stress that is connected with up-regulation of TNF-alpha and its receptors. Also VEGF and VEGF R2 are up-regulated. These responses may be linked to both the damage and repair process of the cochlea. PMID- 15811695 TI - Activation of caspase-3 is associated with oxidative stress in the hydropic guinea pig cochlea. AB - The aim of this study was to investigate the involvement of oxidative stress and apoptosis in an animal model of Meniere's disease. Endolymphatic hydrops (ELH) is generally accepted as the decisive histological characteristic of Meniere's disease. Closure of the endolymphatic duct (Kimura's method) was used to induce endolymphatic hydrops in guinea pigs. Sham-operated animals served as controls. After 4 weeks the animals operated showed a significant elevation of the hearing thresholds as measured by audiometric brainstem responses (ABR) pre- and postoperatively. Immediately after the second ABR measurement, the animals were sacrificed for further immunohistological examinations of the inner ear with specific antibodies to active caspase-3 (cas-3) as a marker for apoptosis and antibodies to 8-isoprostane (8-iso) and nitrotyrosine (NT) as indicators of oxidative stress. Compared with the sham-operated controls, hydropic cochleae showed strong immunostaining for both oxidative stress markers in spiral ganglion cells, in the blood-vessels and fibrocytes of the lateral wall, as well as in supporting cells of the organ of Corti. Activation of cas-3 in spiral ganglion cells and the lateral wall was found exclusively in hydropic cochleae. Our findings suggest that oxidative stress is involved in the development of endolymphatic hydrops and may lead to cellular damage which induces apoptosis by activation of cas-3. Apoptotic cell death might contribute to the sensorineural hearing loss found in later stages of Meniere's disease. PMID- 15811696 TI - Concurrent measurement of the detectability of tone bursts and their effect on the excitability of the human blink reflex using a probe-signal method. AB - The probe-signal method has shown that auditory signals that are either presented more often in a series of trials or that are immediately preceded by cues of the same frequency on a single trial are detected more readily than signals of other frequencies. The frequency range in which detection is favored defines an attentional band, which is thought to result from an effective attenuation of deviant frequencies in the cochlea, possibly by activation of the olivocochlear bundle. In a 2IFC procedure in which the first observation interval was preceded by a 1300-Hz cue, subjects detected cued probe tones (at 1300 Hz) but not uncued probe tones (at 1000 Hz or 1600 Hz) at better than chance levels. Concurrent elicitation of a blink reflex by presentation of an air puff in the first observation interval on a random half of the trials showed that cued probes, but not uncued probes, inhibited the size of the blink reflex. These data show that uncued probes do not enter into the low-level sensory processing in the brainstem which is responsible for reflex modification. This finding is consistent with the view that stimuli whose frequency falls outside an attentional band are excluded at the auditory periphery. PMID- 15811697 TI - Spatial distribution of electrically induced high frequency vibration on basilar membrane. AB - We reported that the electrically evoked basilar membrane (BM) vibration at frequencies above the best frequency (BF) showed a lowest BM velocity magnitude, called a "dip", in the velocity-frequency spectra, indicating a cancellation. In the present study, we measured the high frequency BM motion as functions of the longitudinal and radial locations. Measurements were taken at three longitudinal locations in the first turn and the hook region: 14.9, 15.8 and 16.8 mm from the apex, corresponding to the BFs of 17, 21.3 and 28.0 kHz calculated from Greenwood [J. Acoust. Soc. Am. 87, 2592], and at different radial locations across the width of the BM. It was found that the dip frequency (DF) varied with the longitudinal and radial locations. In the longitudinal direction, the average value of the DF was 49.6, 55.6 and 72.8 kHz, respectively. Thus, the longitudinal distribution of the high frequency BM vibration was correlated with the BF. In the radial direction, there was consistent variation of the response spectrum such that the dip was mainly evident in the pectinate zone of the BM. These results imply that the high frequency BM motion is related to mechanical properties of the cochlear partition, including the outer hair cells (OHCs) themselves. Data also indicate different vibration modes across the width of the organ of Corti. PMID- 15811698 TI - Auditory evoked magnetic fields in relation to bandwidth variations of bandpass noise. AB - Auditory evoked magnetic fields in relation to the bandwidth of bandpass noise were examined by magnetoencephalography (MEG). Pure tone and bandpass noises with center frequencies of 500, 1000 or 2000 Hz were used as the auditory signals. All source signals had the sound pressure level set at 74 dB. The stimulus duration was 0.5 s, with rise and fall ramps of 10 ms. Eight volunteers with normal hearing took part in the study. Auditory evoked fields were recorded using a neuromagnetometer in a magnetically-shielded room. The results showed that the peak amplitude of N1m, which was found above the left and right temporal lobes around 100 ms after the stimulus onset, decreased with increasing bandwidth of the bandpass noise. The latency and estimated equivalent current dipole (ECD) locations of N1m did not show any systematic variation as a function of the bandwidth for any of the center frequencies. PMID- 15811699 TI - Loudness growth in cochlear implants: effect of stimulation rate and electrode configuration. AB - In cochlear implant speech processor design, acoustic amplitudes are mapped to electric currents with the intention of preserving loudness relationships across electrodes. Many parameters may affect the growth of loudness with electrical stimulation. The present study measured the effects of stimulation rate and electrode configuration on loudness growth in six Nucleus-22 cochlear implant users. Loudness balance functions were measured for stimuli that differed in terms of stimulation rate, electrode configuration and electrode location; a 2 alternative, forced-choice adaptive procedure (double-staircase) was used. First, subjects adaptively adjusted the amplitude of a 100-pulse-per-second (pps) pulse train to match the loudness of a 1000-pps standard pulse train. For a range of reference stimulation levels, the loudness of the 100-pps stimulus was matched to that of the 1000-pps standard stimulus; loudness balancing was performed for three electrode pairs [(20,22), (1,3), (1,22)]. The results showed that the loudness balance functions between the 100- and 1000-pps stimulation rates were highly subject-dependent. Some subjects' loudness balance functions were logarithmic, while others' were nearly linear. Loudness balance functions were also measured across electrode locations [(20,22) vs. (1,3)] for two stimulation rates (100, 1000 pps). Results showed that the loudness balance functions between the apical and basal electrode pairs highly depended on the stimulation rate. For all subjects, at the 1000-pps rate, the loudness balance functions between the two electrode locations were nearly linear; however, at the 100-pps rate, the loudness balance function was highly nonlinear in two out of six subjects. These results suggest that, for some cochlear implant patients, low-frequency stimulation may be processed differently at different electrode locations; for these patients, acoustic-to-electric amplitude mapping may need to be sensitive to this place-dependent processing when relatively low stimulation rates are used. PMID- 15811700 TI - A physiological place-frequency map of the cochlea in the CBA/J mouse. AB - Genetically manipulated mice have gained a prominent role in in vivo research on development and function of the auditory system. A prerequisite for the interpretation of normal and abnormal structural and functional features of the inner ear is the exact knowledge of the cochlear place-frequency map. Using a stereotaxic approach to the projection site of the auditory nerve fibers in the cochlear nucleus, we succeeded in labelling physiologically characterized auditory nerve afferents and determined their peripheral innervation site in the cochlea. From the neuronal characteristic frequency (CF) and the innervation site in the organ of Corti a place-frequency map was established for characteristic frequencies between 7.2 and 61.8 kHz, corresponding to locations between 90% and 10% basilar membrane length (base = 0%, apex = 100%, mean length measured under the inner hair cells 5.13 mm). The relation between normalized distance from the base (d) and frequency (kHz) can be described by a simple logarithmic function: d(%) = 156.5-82.5 x log(f), with a slope of 1.25 mm/octave of frequency. The present map, recorded under physiological conditions, differs from earlier maps determined with different methods. The simple logarithmic place-frequency relation found in the mouse indicates that mice are acoustic generalists rather than specialists. PMID- 15811701 TI - Development of the human fetal cochlear nerve: a morphometric study. AB - Ontogenesis of the human peripheral auditory pathway is relatively less explored. While the distal part of the auditory perception apparatus (i.e. the cochlea) received attention, studies on the neural element carrying information to the brainstem (i.e. the cochlear nerve) are scarce. In the present study, axonal differentiation, maturation and myelination of the distal end of the human cochlear nerve (CN) were assessed using light and electron microscopy. Seven human fetuses of 12, 15, 18, 20, 22, 28 and 38 weeks' gestation (WG) were analyzed. Light microscopy revealed nerve fascicles as early as 12 WG, initially arranged loosely but later compacted by 18 WG. Myelinated fibers were clearly detected at 28 WG. Ultrastructurally, at 12 WG developing Schwann cells were present between the thin unmyelinated axons. At 15 WG, the fascicular arrangement was distinct with blood vessels in the perineurium. The maximum number of axons was found at 20 WG, which subsequently reduced to reach the adult level at 22 WG. The myelinated axons in the CN were first observed on the left side at 20 WG, following which the number and proportion of myelinated axons increased until term, incorporating both small and large axons. The right CN lagged behind in maturation. Axon size also increased with age. Thus, the maturation of the human CN commences during the mid-gestation period and produces exuberant axons that are eventually pruned at a time when axons start to myelinate. During this developmental period the human CN maintains maturational asymmetry, the functional consequences of which remain to be elucidated. PMID- 15811702 TI - The influence of genetic variation in oxidative stress genes on human noise susceptibility. AB - Noise induced hearing loss (NIHL) is a complex disease caused by an interaction between genetic and environmental factors. Damage in the cochlea as a result of noise exposure appears to be mediated by reactive oxygen species (ROS). To investigate whether genetic variation in the human protective antioxidant system is associated with high or low susceptibility to NIHL, genetic polymorphisms derived from genes involved in the oxidative stress response were analysed in the 10% most susceptible and 10% most resistant extremes of 1200 Swedish noise exposed workers. The genetic polymorphisms included 2 deletion polymorphisms for the GSTM1 and GSTT1 gene, and 14 SNPs derived from the CAT, SOD, GPX, GSR and GSTP1 genes. No significant differences were found between susceptible and resistant groups, providing no support for a major role of genetic variation of antioxidant enzymes in the susceptibility to NIHL. PMID- 15811703 TI - Tonic mechanosensitivity of outer hair cells after loss of tip links. AB - Tip links - the extracellular connectors between the distal ends of adjacent stereocilia - are essential for the fast mechanical gating of hair-cell transducer channels. Transduction in the absence of tip links was investigated for outer hair cells of the adult guinea-pig cochlea by patch-clamp recordings of the whole-cell current during mechanical stimulation of the hair bundle. Loss of tip links induced by application of BAPTA led to permanently opened transducer channels, as evidenced by a constant inward current, loss of response to sinusoidal mechanical deflection of the hair bundle and block by the open-channel blocker dihydrostreptomycin (100 microM). Step deflection of the hair bundle (200 500 nm) in the inhibitory direction exponentially reduced this current to a constant value with time constant, tau(on), of the order of seconds. The current returned exponentially to the pre-stimulus level with time-constant, tau(off), also of the order of seconds. tau(on) was dependent on the inter-stimulus interval, Deltat, such that reducing this interval below about 40 s resulted in an exponentially faster response. tau(off) was independent of Deltat. Application of the calcium ionophore, ionomycin (10 microM), showed that tau(on) became independent of Deltat after saturating elevation of the intracellular Ca(2+) concentration. Flash-photolytic release of intracellular caged calcium (25-microM NP-EGTA/AM) showed that tau(on) is dependent on intracellular Ca(2+) concentration. These experiments imply an intracellular, calcium-dependent gating mechanism for hair-cell transducer channels. PMID- 15811704 TI - Acute effect of nicotine on auditory gating in smokers and non-smokers. AB - This paper investigates the role of cholinergic mechanisms in auditory gating by assessing the acute effects of nicotine, an acetylcholinomimetic drug, on behavioral and electrophysiological measures of consonant-vowel (CV) discrimination in quiet and in broadband noise (BBN). In a single-blind procedure, categorical boundaries and mismatch negativity (MMN) in two conditions (quiet, BBN) were obtained from 10 non-smokers and 4 smokers with normal hearing under two drug conditions (nicotine, placebo). After the nicotine sessions, plasma tests revealed a subject's nicotine concentration and subjects reported any symptoms. Larger MMN areas and steeper slopes at the boundary were interpreted as reflecting better electrophysiological and behavioral CV discrimination, respectively. Results indicate that, in non-smokers, the effects of nicotine on electrophysiological CV discrimination in quiet increase with an increase in severity of symptoms. Specifically, asymptomatic non-smokers (N = 5) demonstrate little improvement (and sometimes decrements) in performance while symptomatic non-smokers (N = 5) exhibit nicotine-enhanced discrimination, as do smokers. In noise, all subjects demonstrate nicotine-enhanced behavioral and electrophysiological discrimination. Additionally, in noise, smokers exhibit a larger number of measurable categorical boundaries as well as larger MMN areas than non-smokers in both placebo and nicotine sessions. Results are consistent with the hypothesis that nicotinic cholinergic mechanisms play a role in the gating of auditory stimuli. PMID- 15811705 TI - Divergent response properties of layer-V neurons in rat primary auditory cortex. AB - Layer-V pyramidal cells comprise a major output of primary auditory cortex (A1). At least two cell types displaying different morphology, projections and in vitro physiology have been previously identified in layer-V. The focus of the present study was to characterize extracellular receptive field properties of layer-V neurons to determine whether a similar breakdown of responses can be found in vivo. Recordings from 105 layer-V neurons revealed two predominant receptive field types. Thirty-two percent displayed strong excitatory V/U-shaped receptive field maps and spiking patterns with shorter stimulus-driven interspike intervals (ISIs), reminiscent of the bursting cells discussed in the in vitro literature. V/U-shaped maps remained relatively unchanged across the three sequential repetitions of the map run on each neuron. Neurons with V/U-shaped maps were also easily depolarized with extracellular current pulse stimulation. In contrast, 47% of the neurons displayed Complex receptive field maps characterized by weak and/or inconsistent excitatory regions and were difficult to depolarize with current pulses. These findings suggest that V/U-shaped receptive fields could correspond to previously described intrinsic bursting (IB) cells with corticotectal projections, and that neurons with Complex receptive fields might represent the regular spiking (RS) cells with their greater inhibitory input and corticocortical/corticostriatal projection pattern. PMID- 15811706 TI - Effects of acoustic noise on the auditory nerve compound action potentials evoked by electric pulse trains. AB - This study investigated the effects of acoustic noise on the auditory nerve compound action potentials in response to electric pulse trains. Subjects were adult guinea pigs, implanted with a minimally invasive electrode to preserve acoustic sensitivity. Electrically evoked compound action potentials (ECAP) were recorded from the auditory nerve trunk in response to electric pulse trains both during and after the presentation of acoustic white noise. Simultaneously presented acoustic noise produced a decrease in ECAP amplitude. The effect of the acoustic masker on the electric probe was greatest at the onset of the acoustic stimulus and it was followed by a partial recovery of the ECAP amplitude. Following cessation of the acoustic noise, ECAP amplitude recovered over a period of approximately 100-200 ms. The effects of the acoustic noise were more prominent at lower electric pulse rates (interpulse intervals of 3 ms and higher). At higher pulse rates, the ECAP adaptation to the electric pulse train alone was larger and the acoustic noise, when presented, produced little additional effect. The observed effects of noise on ECAP were the greatest at high electric stimulus levels and, for a particular electric stimulus level, at high acoustic noise levels. PMID- 15811707 TI - Dexamethasone inhibits tumor necrosis factor-alpha-induced cytokine secretion from spiral ligament fibrocytes. AB - To investigate the effect of proinflammatory cytokines on spiral ligament (SL) fibrocytes and regulation of cytokines by dexamethasone (Dex), in vitro studies were performed in murine secondary cell cultures. Cultured SL fibrocytes were stimulated with tumor necrosis factor-alpha (TNF-alpha), and the secretion of various mediators was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcribed-polymerase chain reaction (RT-PCR). After stimulation with TNF-alpha, levels of keratinocyte-derived cytokine (KC), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), interleukin-6 (IL 6) and soluble intercellular adhesion molecule-1 (sICAM-1) were elevated in the culture supernatant, and their corresponding messenger RNAs were detected in the cultured fibrocytes. When the cultures were incubated with both TNF-alpha and Dex, the levels of KC, MCP-1, MIP-2 and IL-6 were significantly lower than those in cultures treated with TNF-alpha alone. The data suggest that Dex suppresses the inflammatory response in SL fibrocytes. Given that SL fibrocytes play a role in cochlear fluid and ion homeostasis, glucocorticoids may suppress the cochlear malfunction caused by SL inflammation. PMID- 15811709 TI - Current-level discrimination using bipolar and monopolar electrode configurations in cochlear implants. AB - This study examined current-level discrimination ability in listeners with cochlear implants using bipolar and monopolar electrode configurations. Current level discrimination ability was measured as a function of electrode configuration (monopolar and bipolar), stimulation site (8 and 16) and level (5%, 15%, 25%, 50% and 80% of the dynamic range). Weber fractions usually decreased with increasing level. Differences in Weber fractions between monopolar and bipolar configurations were observed for listeners with short durations of deafness (<5 years). For these listeners, in the bipolar condition at the more apical site 16, Weber fractions remained constant with increasing level, and the Weber fractions at low levels were smaller than in other conditions. We suggest that nerve density was better and the nerve-to-site-of-action-potential distance was smaller in these cases such that more fibers could be recruited with a unit increase in current level, leading to better current-level sensitivity. PMID- 15811708 TI - Characterization of a new allele of Ames waltzer generated by ENU mutagenesis. AB - Mutation in the protocadherin 15 (Pcdh15) gene causes hair cell dysfunction and is associated with abnormal stereocilia development. We have characterized the first allele (Pcdh15(av-nmf19)) of Ames waltzer (av) obtained by N-ethyl-N nitrosourea (ENU) mutagenesis. Pcdh15(av-nmf19) was generated in the Neuroscience Mutagenesis Facility (NMF) at The Jackson Lab (Bar Habor, USA). Pcdh15(av-nmf19) mutants display circling and abnormal swimming behavior along with lack of auditory-evoked brainstem response at the highest intensities tested. Mutation analysis shows base substitution (A--> G) in the consensus splice donor sequence linked to exon 14 resulting in the skipping of exon 14 and the splicing of exon 13-15. This results in the introduction of a stop codon in the coding sequence of exon 15 due to shift in the reading frame. The effect of nmf19 mutation is expected to be severe since the expressed Pcdh15 protein is predicted to truncate in the 5th cadherin domain. Abnormalities of cochlear hair cell stereocilia are apparent in Pcdh15(av-nmf19) mutants near the time of birth and by about P15 (15 days after birth) there is evidence of sensory cell degeneration. Disorganization of outer hair cell stereocilia is observed as early as P2. Inner hair cell stereocilia are also affected, but less severely than those of the outer hair cells. These results are consistent with characteristics of the mutation in the Pcdh15(av-nmf19) allele and they support our previous finding that Protocadherin 15 plays an important role in hair-bundle morphogenesis. PMID- 15811710 TI - Morphology of the endolymphatic sac in the guinea pig after an acute endolymphatic hydrops. AB - The role of the endolymphatic sac (ES) in endolymph volume homeostasis is speculative. The present study investigates changes of the ES's epithelia and luminal filling after induction of an acute endolymphatic hydrops. After microinjection of 1.1 mul artificial endolymph into scala media of the cochlea, guinea pigs were terminated immediately (n = 6) or after different time intervals ; 1/2 h (n = 3), 1 h (n = 4) and 2 h (n = 4). Inner ear specimens were processed for light and/or transmission electron microscopy. The non-injected contralateral ear served as a histological control. Correct injection was confirmed by detection of microspheres in the endolymphatic compartment after the same microinjection procedure. In all specimens, ribosome rich cells and intraluminal macrophages appeared to be actively involved in degradation of homogeneous substance (HS) by secreting lytic enzymes and digestion, respectively. Amazingly, in our study no ES differences were found between injected and non-injected ears and no distinct changes were observed in guinea pigs terminated after different time intervals. The ES's luminal HS was always present and often to a large extent. This is in contrast with [Hear. Res. 138, 81] dramatic changes were observed. Endolymph volume homeostasis is a complex mechanism, in which the role of HS remains obscure. PMID- 15811711 TI - Psychophysical evidence for adaptation of central auditory processors for interaural differences in time and level. AB - Human listeners were studied for their ability to lateralize single target tones of each of two frequencies relative to midline clicks. They did so before and after exposure to adaptor tones of the same frequencies. The adaptor tones were strongly lateralized, and in opposite directions for each frequency, by either an interaural time difference (ITD, Experiment 1) or interaural level difference (ILD, Experiment 2). Following adaptation, psychometric functions for ITD (Exp. 1) and ILD (Exp. 2) were obtained for target tones for the two frequencies separately. These were found to be shifted in the direction of the fatigued side. In the case of ILD, this was in the absence of a shift in monaural sensitivity sufficient to account for the effect. For both ITD and ILD studies, shifts in perceived laterality were induced in opposite directions at two frequencies concurrently. This effect was induced with only seconds of intermittent exposure to the adaptor tones. The fact that it could be induced at two frequencies in opposite directions at the same time, suggests (a), that these data constitute new psychophysical evidence for the frequency specificity of ITD and ILD coding in the human brain, and (b), that the effect was not due to the introduction of some response bias at the decision level of perceptual judgement. The data are interpreted in terms of a two- or three-channel opponent process model. PMID- 15811712 TI - Dietary vitamin C supplementation reduces noise-induced hearing loss in guinea pigs. AB - Vitamin C (ascorbate) is a water-soluble, low molecular weight antioxidant that works in conjunction with glutathione and other cellular antioxidants, and is effective against a variety of reactive oxygen species, including superoxide and hydroxyl radicals that have been implicated in the etiology of noise-induced hearing loss (NIHL). Whereas most animals can manufacture their own vitamin C, humans and a few other mammals such as guinea pigs lack the terminal enzyme for vitamin C synthesis and must obtain it from dietary sources. To determine if susceptibility to NIHL could be influenced by manipulating dietary levels of vitamin C, albino guinea pigs were raised for 35 days on a diet with normal, supplemented or deficient levels of ascorbate, then exposed to 4 kHz octave band noise at 114 dB SPL for 6 h to induce permanent threshold shifts (PTS) of the scalp-recorded auditory brainstem response. Animals that received the highest levels of dietary ascorbate developed significantly less PTS for click stimuli and 4, 8, 12, and 16 kHz tones than animals on normal and deficient diets. Outer hair cell loss was minimal in all groups after noise exposure, but permanent damage to stereocilia were observed in noise-exposed ears. The results support the hypothesis that dietary factors influence individual susceptibility to hearing loss, and suggest that high levels of vitamin C may be beneficial in reducing susceptibility to NIHL. PMID- 15811713 TI - Effect of cochlear integrity on cochlear nucleus neuron glucose metabolism in aged adult broiler chickens. AB - Abrupt removal of excitatory input is devastating to post-synaptic neurons in normally functioning sensory systems. In both mammalian and avian auditory systems, abrupt temporary or permanent experimental deafferentation stimulates a cascade of changes in central auditory structures that can result in neuron death. Effects of naturally occurring progressive deafferentation on central auditory structure and function have not been fully described. Extensive naturally occurring cochlear damage is found in some aged chickens, despite their regenerative capacity, providing the opportunity to examine the effects of this type of deafferentation on the avian cochlear nucleus (nucleus magnocellularis, NM). Previous evaluation of NM oxidative metabolism using cytochrome oxidase histochemistry revealed that naturally occurring cochlear damage results in down regulated metabolism in corresponding regions of NM. It is unknown how progressive hair cell damage and loss affects NM glucose uptake. Here, NM glucose metabolism is assessed using 2-deoxyglucose uptake as a marker for metabolic activity in the presence of normal, mildly damaged, severely damaged, and totally damaged cochlear hair cells. Results indicate that while severe and total cochlear damage significantly decrease NM oxidative metabolism, only total damage results in significantly decreased NM glucose metabolism. Results are discussed in the context of functional reorganization and trophic support. PMID- 15811714 TI - The role of GABAergic inhibition in shaping the response size and duration selectivity of bat inferior collicular neurons to sound pulses in rapid sequences. AB - Natural sounds, such as vocal communication sounds of many animal species typically occur as sequential sound pulses. Therefore, the response size of auditory neurons to a sound pulse would be inevitably affected when the sound pulse is preceded and succeeded by another sound pulse (i.e., forward and backward masking). The present study presents data to show that increasing strength of GABAergic inhibition relative to excitation contributes to decreasing response size and sharpening of duration selectivity of bat inferior collicular (IC) neurons to sound pulses in rapid sequences. The response size in number of impulses and duration selectivity of IC neurons were studied with a pulse train containing 9 sound pulses. A family of duration tuning curves was plotted for IC neurons using the number of impulses discharged to each presented sound pulse against pulse duration. Our data show that the response size of IC neurons progressively decreased and duration selectivity increased when determined with sequentially presented sound pulses. This variation in the response size and duration selectivity of IC neurons with sequentially presented sound pulses was abolished or reduced during bicuculline and GABA application. Bicuculline application increased the response size and broadened the duration tuning curve of IC neurons while GABA application produced opposite results. Possible mechanisms underlying increasing strength of GABAergic inhibition with sequentially presented sound pulses are presented. Biological significance of these findings in relation to acoustic signal processing is also discussed. PMID- 15811715 TI - Attribute capture in the precedence effect for long-duration noise sounds. AB - Listeners perceptually fuse the direct wave from a sound source with its reflections off nearby surfaces into a single sound image, located at or near the sound source (the precedence effect). This study investigated how a brief gap presented in the middle of either a direct wave or simulated reflection is incorporated into the fused image. For short (<9.5 ms) delays between the direct (leading) and reflected (lagging) waves, no sound was perceived from the direction of the lagging wave. For delays between 10 and 15 ms, both sounds were perceived, but the gap was heard only on the leading side. When the gap was only in the correlated lagging sound at short delays, it also was perceived as occurring on the leading side. Moreover, gap detection thresholds were the same for gaps in the leading and lagging sounds, suggesting that the perception of the gap was not suppressed, but rather incorporated into the leading sound. Finally, scalp event-related potentials were not associated with the precedence effect until the gap occurred. This suggests that cortical mechanisms are engaged to maintain fusion when attributes in direct or reflected waves change. PMID- 15811716 TI - Time-intensity trading in bilateral congenital aural atresia patients. AB - In an effort to examine the rules by which information of bilaterally applied bone-conducted signals arising from interaural time differences (ITD) and interaural intensity differences (IID) is combined, data were measured for continuous 500 Hz narrow band noise at 65-70 dB HL in 11 patients with bilateral congenital aural atresia. Time-intensity trading functions were obtained by shifting the sound image towards one side using ITD, and shifting back to a centered sound image by varying the IID in the same ear (auditory midline task). ITD values were varied from -600 to +600 micros at 200 micros steps, where negative values indicate delays to the right ear. The results indicate that time intensity trading is present in patients with bilateral aural atresia. The gross response properties of time-intensity trading in response to bone-conducted signals were comparable in patients with bilateral aural atresia and normal hearing subjects, though there was a larger inter-subject variability and higher discrimination thresholds across IIDs in the atresia group. These results suggest that the mature auditory brainstem has a potential to employ binaural cues later in life, although to a restricted degree. A binaural fitting of a bone-conducted hearing aid might optimize binaural hearing and improve sound lateralization, and we recommend now systematically bilateral fitting in aural atresia patients. PMID- 15811717 TI - A novel missense mutation in the Connexin 26 gene associated with autosomal recessive sensorineural deafness. AB - Mutations in the Connexin 26 (Cx26) gene (GJB2) are a common cause of hereditary hearing impairment. We report the identification of a novel point mutation in the Cx26 gene, Leu205Pro(L205P), linked to familial, autosomal recessive sensorineural hearing loss. This missense mutation, causing amino acid leucine at position 205 to be substituted by proline, is located in the highly conserved sequence of the fourth transmembrane domain (TM4) of Cx26. Hearing loss with this mutation occurred in a Georgian Jewish family, was congenital, moderate to profound and nonprogressive. We have shown that the new mutation L205P in Cx26 is strongly associated with congenital NSHL. Multiple-sample screening for this mutation can be easily performed with a mismatch PCR that creates a restriction site. PMID- 15811719 TI - Duration of pupillary constriction in response to a photographic flash. PMID- 15811721 TI - Improvement in cognitive impairment after cataract surgery in elderly patients. PMID- 15811722 TI - Assessment of Pascal dynamic contour tonometer in monitoring IOP after LASIK. PMID- 15811724 TI - Effect of 1-piece and 3-piece AcrySof IOLs on the development of PCO after cataract surgery. PMID- 15811725 TI - Lens-iris diaphragm retropulsion syndrome during phacoaspiration in pediatric cataracts. PMID- 15811726 TI - Consultation section refractive surgical problem. PMID- 15811732 TI - Closed foldable capsular rings. AB - A new foldable capsular tension and bending ring system with a sharp-edged design is described. The closed foldable capsular rings (CFCR) consist of 8 hydrophobic and 8 hydrophilic ring segments. The CFCRs have a 9.2 mm minimum overall diameter. The CFCRs were inserted using various cartridge systems or a two-folded technique with a forceps. The CFCRs were implanted after phacoemulsification through a small (1.6 to 3.2 mm) incision in 104 human adult eyes without intraoperative complications. No postoperative complications such as capsule folds or inward bending were observed over a 6-month follow-up. Posterior capsule opacification was minimal or absent in all eyes. PMID- 15811733 TI - Combined pars plana vitrectomy and phacoemulsification to restore visual acuity in patients with chronic uveitis. AB - PURPOSE: To report the outcomes of combined phacoemulsification and pars plana vitrectomy (PPV) to restore visual acuity in patients with cataract and posterior segment involvement secondary to chronic uveitis. SETTING: Ocular Immunology and Uveitis Service, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, USA. METHODS: This study comprised 34 patients (20 women, 14 men; 36 eyes) with posterior segment involvement secondary to chronic uveitis who had combined phacoemulsification and PPV from 1998 to 2002. The main outcome measures were visual acuity, intraocular pressure, and cystoid macular edema. RESULTS: The mean patient age was 45 years +/- 16.09 (SD). The mean duration of uveitis before surgery was 56 +/- 44.17 months. In 24 eyes (66.7%), an intraocular lens (IOL) was implanted during surgery; 12 eyes (33.3%) were left aphakic. Five eyes (13.8%) received an intraocular steroid injection intraoperatively. Visual acuity improved in 26 eyes (72.2%), deteriorated in 5 (13.9%), and was unchanged in 5 (13.9%). The main reason for decreased visual acuity was refractory macular edema. During the follow-up, 2 IOLs were explanted secondary to lens intolerance. One IOL was repositioned because of iris capture by the haptics, and 1 dislocated inferiorly, causing monocular diplopia. The mean follow-up was 23.4 +/- 16.7 months. CONCLUSIONS: Results indicate that combined phacoemulsification and PPV is a feasible technique for the removal of cataract and pathologic vitreous in eyes with chronic uveitis. Although the exact role of vitrectomy in patients with uveitis remains to be determined, the combined surgery successfully restored useful vision in most cases. PMID- 15811734 TI - Long-term intraocular pressure control after clear corneal phacoemulsification in glaucoma patients. AB - PURPOSE: To evaluate long-term IOP control after sutureless clear corneal phacoemulsification in eyes with preoperatively controlled glaucoma. SETTING: Institutional study. METHODS: The charts of 345 patients who had uneventful sutureless clear corneal phacoemulsification with acrylic foldable lens (IOL) implantation were retrospectively reviewed. Included were 58 patients with medically controlled open-angle glaucoma and 287 normal controls. Follow-up was 1 to 2 years. Outcome measures were postoperative IOP and number of glaucoma medications. RESULTS: Postoperatively, there was an insignificant decrease in IOP in the glaucoma group; the mean decrease was 1.5 mm Hg +/- 4.4 (SD) at 12 months and 1.9 +/- 4.9 mm Hg at 24 months. The mean number of medications decreased significantly at 12 months (0.53 +/- 0.86) and at 24 months (0.38 +/- 0.9) (P=.04). The control group also had a significant decrease in IOP, with a mean decrease of 0.72 +/- 3.7 mm Hg at 12 months (P=.01) and 1.33 +/- 3.2 mm Hg at 24 months (P<.0001). The decrease in IOP was more pronounced in eyes with a higher preoperative IOP in both the glaucoma and control groups. CONCLUSIONS: These findings suggest that sutureless clear corneal phacoemulsification with foldable acrylic IOL implantation is a relatively simple and efficient surgical option in patients with cataract and well-controlled glaucoma. The approach combines long term IOP control with fewer medications and leads to rapid visual rehabilitation. PMID- 15811735 TI - Medical control of intraocular pressure after phacoemulsification. AB - PURPOSE: To compare the effectiveness of oral acetazolamide, topical brinzolamide 1%, and no ocular hypotensive medication after phacoemulsification. SETTING: Adnan Menderes University Department of Ophthalmology, Aydin, Turkey. METHODS: This prospective randomized double-blind study comprised 60 eyes of 52 patients having phacoemulsification under topical anesthesia. There were no intraoperative complications. Eyes were randomized to receive oral acetazolamide 500 mg 1 hour preoperatively followed by 250 mg acetazolamide every 6 hours, 1 drop of brinzolamide 1% every 12 hours starting immediately after speculum removal, or no ocular hypotensive medication. Intraocular pressure (IOP) was measured using a Perkins tonometer preoperatively and 4 to 6 hours and 18 to 24 hours postoperatively. RESULTS: The preoperative IOP was not significantly different between the 3 groups. Four to 6 hours postoperatively, the acetazolamide group (P=.002) and brinzolamide group (P=.001) had significantly lower IOP than the control group. The same trend was observed at 18 to 24 hours in the brinzolamide group (P=.001) but not the acetazolamide group (P=.018). The IOP levels were not significantly different between the acetazolamide group and brinzolamide group at any postoperative time point. No eye receiving medication and 2 eyes (10%) in the control group had an IOP of 30 mm Hg or higher 4 to 6 hours postoperatively. Compared with preoperatively, an IOP increase of more than 5 mm Hg was seen at 4 to 6 hours in 3 eyes (15%), 2 eyes (10%), and 14 eyes (70%) in the acetazolamide, brinzolamide, and control group, respectively. CONCLUSION: Brinzolamide was as effective as acetazolamide in preventing IOP elevation 4 to 6 hours after phacoemulsification and more effective than acetazolamide at 18 to 24 hours. PMID- 15811736 TI - Off-axis refraction and aberrations following conventional laser in situ keratomileusis. AB - PURPOSE: To investigate off-axis refraction and aberrations following conventional laser in situ keratomileusis (LASIK) for myopia and hypermetropia. SETTING: School of Optometry, Queensland University of Technology, Australia. METHODS: Using an autorefractor, off-axis refractions were analyzed along the horizontal visual field between 35 degrees nasally and 35 degrees temporally in 1 eye each of 15 emmetropic subjects (-0.50 to +0.50 diopters [D]), 6 myopic subjects (-2.25 to -6.50 D), 6 hyperopic subjects (+1.50 to +3.00 D), 6 myopic LASIK patients (presurgical refraction -2.75 to -9.00 D), and 6 hyperopic LASIK patients (presurgical refraction +0.75 to +2.00 D). Wavefront sensing measured off-axis higher-order aberrations in 2 myopic LASIK patients. RESULTS: In myopic LASIK, the mean spherical components of refraction M became highly myopic away from the center of the visual field; in emmetropic and untreated myopic eyes, there were relatively small myopic shifts and hyperopic shifts, respectively. Off axis 90-degree to 180-degree astigmatisms J180 in myopic LASIK subjects were greater than in untreated subjects. In hyperopic LASIK, there were mainly hyperopic shifts in M, opposite the direction in emmetropic and untreated hyperopic subjects. Off-axis J180 was less than in emmetropic and untreated hyperopic subjects. Some hyperopic LASIK patients had greater off-axis 45-degree to 135-degree astigmatisms J45 than patients in the other groups. In 2 myopic LASIK patients, Zernike root-mean-square 4th-order aberrations were higher than in the near-emmetropia group because of higher levels of positive spherical aberration. CONCLUSIONS: Off-axis aberrations can be dramatically affected by conventional myopic and hyperopic LASIK. In myopic LASIK, the increased off-axis refractive errors may have adverse effects on peripheral visual tasks that are dependent on off-axis refractive errors. The relatively low off-axis refractive errors in hyperopic LASIK patients may improve peripheral visual tasks. PMID- 15811737 TI - Posterior assisted levitation: long-term follow-up data. AB - PURPOSE: To report the long-term outcome of posterior assisted levitation (PAL) in cases of a sinking dropped nucleus. SETTING: Department of Ophthalmology, Soroka University Medical Center, Beer-Sheva, Israel. METHODS: The medical records of all the patients having cataract surgery at Soroka University Medical Center from January 1999 to December 2002 were retrospectively reviewed. The PAL technique was used in 7 eyes, 3 of which had mini-nuc cataract surgery and 4 of which had phacoemulsification. Posterior assisted levitation was done because of an intraoperative posterior capsule tear with vitreous loss and a sinking dropped nucleus. A spatula was inserted via the pars plana, and the nucleus was lifted to and removed from the anterior chamber. RESULTS: The follow-up ranged from 16 to 28 months. The postoperative visual acuity was 20/40 or better in all but 2 eyes, which had 20/50 acuity because of age-related macular degeneration and diabetic macular edema, respectively. No postoperative complications were observed. CONCLUSIONS: The PAL technique was successfully performed during cataract surgery in 7 eyes with a posterior capsule tear, vitreous loss, and a sinking dropped nucleus. There were no complications over the long-term follow-up. PMID- 15811738 TI - Results of late flap removal after complicated laser in situ keratomileusis. AB - PURPOSE: To evaluate the effect of flap removal on complications after laser in situ keratomileusis (LASIK). SETTING: Three university-based referral centers and 1 private practice. METHODS: This retrospective interventional case series comprised 6 eyes of 6 patients at 4 centers. Flap removal occurred 2 to 41 weeks after the LASIK procedure. The corneal flaps were excised by 2 methods: In 2 eyes, the flap was lifted and excised manually. In 4 eyes, the thin flap was removed by excimer ablation using phototherapeutic keratectomy and/or photorefractive keratectomy. Postoperative measurements included uncorrected visual acuity, best spectacle-corrected visual acuity (BSCVA), manifest refraction, slitlamp evaluation, and computerized videokeratography. All patients had an 8-month or longer convalescence to assess visual recovery. RESULTS: After the initial flap complication, the BSCVA decreased in all 6 eyes (mean loss 3.0 lines +/- 1.5 [SD]). After flap removal, it improved in all eyes (mean gain 2.2 +/- 1.2 lines). All patients reported a reduction in or elimination of visual symptoms. Despite the improvements, a minor loss of BSCVA (mean -0.8 lines [range 0 to 2 lines]) remained in 4 patients. CONCLUSION: In carefully selected patients, flap removal is a viable surgical option to improve visual function. PMID- 15811739 TI - Traumatic cataract and other adverse events with the implantable contact lens. AB - PURPOSE: To evaluate the surgical outcome and adverse events associated with implantation of the implantable contact lens (ICL) for the correction of high myopia or high hyperopia. SETTING: Helsinki University Eye Hospital, Helsinki, Finland. METHODS: In this nonrandomized prospective clinical trial, the ICL V4 was implanted in 26 eyes of 13 patients with normal enrollment criteria (younger than 45 years, a clinically clear crystalline lens) (Group 1) and the V2, V3, or V4 were implanted in 38 eyes of 22 patients with heterogeneous criteria (older than 45 years or opacities in the crystalline lens) (Group 2). The main outcome measures were lens opacity classification system (LOCS II) assessment and transparency analysis of the lens. RESULTS: In Group 1, microdensitometric assessment showed no decrease in crystalline lens transparency and no statistically significant increase in crystalline lens density at any follow-up examination (P>.05). The incidence of anterior subcapsular (AS) opacities was 7.7%, and no eye developed clinically significant cataract during the mean follow up of 13.2 months +/- 12.3 (SD). No myopic eye lost a line of best corrected visual acuity (BCVA); 50.0% of hyperopic eyes lost 1 line of BCVA. Sixty-eight percent of myopic eyes and 75.0% of hyperopic eyes were within +/-1.0 diopter of the predicted refraction. In Group 2, the incidence of AS opacities was 47.4% and 10 eyes (26.3%) developed clinically significant cataract during the mean follow up of 30.9 +/- 18.0 months. The visual and refractive results were similar to those in Group 1 at 1 month, but due to progressive AS opacities in the crystalline lens, 23.4% of eyes lost BCVA lines during the follow-up. Intraoperative complications were few and insignificant. CONCLUSIONS: Implantation of the ICL for correction of high myopia or high hyperopia appears to be a safe procedure with good visual and refractive results immediately. There was no decrease in the transparency or increase in the density of the crystalline lens with the latest generation of the ICL device in the normal group, indicating enhanced safety. Progressive lens opacification in the heterogeneous group shows there are risk factors for cataract formation; ie, type V3 ICL, preexisting lens opacities, and older patient age. Anterior subcapsular opacities are most likely the result of trauma. PMID- 15811740 TI - Topography-based intraocular lens power selection. AB - PURPOSE: To provide mathematical tools for selecting intraocular lens (IOL) power for normal eyes and for "odd" eyes, particularly after corneal refractive surgery. SETTING: Universitats-Augenklinik, Mainz, Germany. METHODS: First, IOL power is selected based on the radii and numerical eccentricity of the cornea, extracted from corneal topography in a consistent numerical model of the cornea. To fine-tune the result, the visual impression is simulated by blurred Landolt rings superimposed on the retinal receptor grid. The calculation uses numerical ray tracing of the whole pseudophakic eye comprising all monochromatic errors. The error contributions of the influencing parameters, such as anterior and posterior corneal shape and corneal thickness, are quantified in detail. The method is verified in IOL power selection for normal eyes and for eyes after corneal refractive surgery. RESULTS: The main difference between normal corneas and corneas after refractive surgery results from different asphericities. Normal corneas are prolate, with typical numerical eccentricities of 0.5, whereas corneas after laser surgery for myopia are oblate. This causes the main difference (hyperopic shift up to 2.0 diopters) in IOL power selection. Shifts in the posterior corneal radius and corneal thickness are of minor importance. CONCLUSION: Intraocular power selection after corneal refractive surgery should be based on all the information corneal topography provides. PMID- 15811741 TI - Correlation between the measurement of posterior capsule opacification severity and visual function testing. AB - PURPOSE: To develop software to measure the severity of posterior capsule opacification (PCO) using analysis of retroillumination images and to correlate the results with clinical evaluation of PCO severity and visual function. SETTING: Department of Physics, King's College, and Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: A technique for calculating PCO severity was developed based on calculating the variance of intensity by transforming retroillumination images to a similar mean intensity. The computer-derived severity was compared to grading of clinical severity by 3 independent observers using a library of 100 retroillumination images ranging from clear posterior capsules to very severe PCO. The computer results were also compared with the following other current methods of measuring PCO: Evaluation of Posterior Capsule Opacification (EPCO), POCOman, and Automated Quantification of After-Cataract. A further 35 images were used to compare the results of computer derived severity with the results of visual function analysis (high-contrast acuity, 100%; low-contrast acuity, 9%) using the Early Treatment Diabetic Retinopathy Study chart, contrast sensitivity testing using the Pelli-Robson chart, and glare assessment using the van den Berg straylight meter. RESULTS: The severity scores showed a good correlation with clinical severity scores for the library of images (r=0.86) and with severity scores using POCOman and EPCO (r=0.85 and r=0.81, respectively). The correlations with visual function tests were also good, with low-contrast visual acuity (9%) showing the best correlation (r=0.87). CONCLUSION: Variance in intensity of PCO was successfully used to calculate the severity of PCO. PMID- 15811742 TI - Association between intensity of posterior capsule opacification and visual acuity. AB - PURPOSE: To investigate the correlation between the change in visual acuity and the difference in objective posterior capsule opacification (PCO) scores before and after neodymium:YAG (Nd:YAG) laser capsulotomy. SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: Forty pseudophakic eyes of 35 patients with PCO of varying intensity were examined before and after Nd:YAG laser capsulotomy. Visual acuity was determined using the Early Treatment Diabetic Retinopathy Study (ETDRS) chart at 4 m and the Holladay reading chart at 40 cm. The pupil diameter under reading conditions was measured each time. Digital retroillumination images of the posterior capsule were taken, and the corresponding area inside the pupil was evaluated using the Automated Quantification of After-Cataract (AQUA) automated PCO analysis program. The change in visual acuity and difference between PCO scores before and after Nd:YAG laser capsulotomy were calculated for all eyes. RESULTS: The mean AQUA score (scale 0 to 10) was 3.56 before and 0.13 after Nd:YAG laser capsulotomy. The mean ETDRS visual acuity score (logMAR scale) was 0.28 and -0.07, respectively. The correlation coefficient between the differences in PCO score and distance visual acuity was 0.61 and near visual acuity, 0.62. CONCLUSIONS: The objective PCO score obtained by an automated image-analysis program correlates well with the PCO-induced decrease in visual acuity when the central area (inside the pupillary aperture) of the posterior lens capsule was evaluated. Objective PCO assessment by automated image-analysis systems is, therefore, a valuable and clinically relevant method for clinical studies of the development and prevention of PCO. PMID- 15811743 TI - Comparison of the potential acuity meter and pinhole tests in predicting postoperative visual acuity after cataract surgery. AB - PURPOSE: To compare the accuracy of potential acuity meter (PAM) and pinhole (PH) tests in predicting visual acuity after cataract surgery. SETTING: Department of Ophthalmology and Visual Sciences, University of the Philippines, Philippine General Hospital, Manila, and Asian Eye Institute, Makati, Philippines. METHODS: This prospective study comprised 64 eyes with mild to moderate cataract that had uneventful phacoemulsification. The PAM and PH tests were performed to predict postoperative visual acuity. Best corrected visual acuity (BCVA) 4 weeks after surgery was compared with the predicted visual acuity. The number of lines of inaccuracy was calculated by subtracting the BCVA from the predicted visual acuity. The variables analyzed were type of predictive test and preoperative BCVA. The eyes were divided according to preoperative BCVA as follows: Group 1, 20/20 to 20/50; Group 2, 20/60 to 20/100; Group 3, 20/200 or worse. RESULTS: The PH predicted visual acuity was correct in 5% of eyes and the PAM predicted acuity, in 17%. The PH predicted acuity was accurate within 1, 2, and 3 lines of BCVA in 23%, 40%, and 54% of eyes, respectively, and the PAM predicted acuity, in 64%, 81%, and 92% of eyes, respectively. The mean number of lines of inaccuracy was significantly less with the PH than with the PAM (3.47 lines +/- 2.42 [SD] and 1.60 +/- 1.55 lines, respectively) (P=.0005). The mean lines of inaccuracy in Group 1 were 2.49 +/- 1.52 for the PH and 1.14 +/- 0.99 for the PAM (P=.027); in Group 2, 3.17 +/- 1.99 PH and 1.65 +/- 1.80 PAM (P=.642); and in Group 3, 6.58 +/ 3.03 PH and 2.67 +/- 2.10 PAM (P=.240). CONCLUSIONS: The PAM was more accurate than the PH in predicting visual acuity after cataract surgery. The accuracy of both tests decreased in patients with poorer preoperative visual acuity. PMID- 15811744 TI - Pupil response to tropicamide following laser in situ keratomileusis. AB - PURPOSE: To investigate the effect of corneal thinning after laser in situ keratomileusis (LASIK) on the corneal penetration of topical eye medication. SETTING: Department of Ophthalmology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA. METHODS: Laser in situ keratomileusis surgery was performed in 19 eyes of 10 patients enrolled in this prospective study. Measurements were made before surgery and 3 months postoperatively. After instillation of tropicamide 1%, the change in pupil size over time was measured with a Colvard pupillometer. Central corneal thickness (CCT) was measured with ultrasonic pachymetry before and 3 months after LASIK. The corneal epithelial condition was also examined by fluorescein dye staining. RESULTS: The mean CCT decreased significantly from 564 microm +/- 33 (SD) before LASIK surgery to 514 +/- 48 microm 3 months postoperatively (P<.0001). Pupil diameter 10, 15, and 20 minutes after tropicamide 1% instillation was significantly larger 3 months after surgery than preoperatively (P=.0083, P=.0043, and P=.0144, respectively). The mean time to reach a pupil diameter of 6.0 mm decreased significantly from 14.4 +/- 4.3 minutes in preoperative eyes to 11.5 +/- 2.3 minutes in postoperative eyes (P=.0281). Mild punctate corneal epithelial staining (fewer than 5 spots) were observed in 4 eyes at the 3-month postoperative examination. CONCLUSIONS: Pupil dilation after tropicamide 1% instillation was significantly faster after LASIK surgery. Corneal thinning that resulted from LASIK enhanced corneal penetration of tropicamide 1%. PMID- 15811745 TI - Evaluating the eye's rotational stability during standard photography: effect on determining the axial orientation of toric intraocular lenses. AB - PURPOSE: To evaluate the rotational stability of the eye during standard photography and determine its effect on the measurement of toric intraocular lens (IOL) orientation. SETTING: Department of Ophthalmology, University Erlangen Nuremberg, Erlangen, Germany. METHODS: The rotational stability of the eye was evaluated using standard photographs taken with a telecentric fundus camera (Zeiss). Two sets of fundus images were taken at least 6 months apart in 400 eyes of 200 patients. The axial position of the eye was determined using 2 characteristic markers of the fundus. The angle between the 2 images (autorotation angle) was measured in each eye. RESULTS: The mean absolute autorotation was 2.3 degrees +/- 1.7 (SD) (range 0 to 11.5 degrees). Nine percent of eyes did not rotate. The rotation was less than 3 degrees in 55% of eyes and was 3 degrees or greater in 36% of eyes. Eyes of patients younger than 50 years rotated less than eyes in older patients (mean 2.2 +/- 1.5 degrees and 2.5 +/- 1.8 degrees, respectively) (P=.04). A visual acuity of 20/20 or better (P=.02) and a refractive cylinder of less than 1.75 diopters (P=.01) were correlated with smaller amounts of autorotation. Potential causes of artificial eye rotation induced by the photographic technique included camera adaptation (3-degree intrinsic error), slide mounting (<1 degree), slide projection (<0.5 degree), marking of characteristic fundus details (<1 degree), and head inclination. CONCLUSIONS: Cyclorotation of the eye during standard photography may lead to overestimation or underestimation of the presumed spontaneous rotation of an implanted toric IOL. Results show that 11.5 degrees of toric IOL rotation would lead to residual astigmatism that is 40% of the initial astigmatic power and 3 degrees, 10% of the initial power. Digital imaging may reduce the intrinsic errors of standard photography. PMID- 15811746 TI - Intraocular lens calculations after refractive surgery. AB - PURPOSE: To evaluate the effect of refractive surgery on intraocular lens (IOL) power calculation, compare methods of IOL power calculation after refractive surgery, evaluate the effect of pre-refractive surgery refractive error on IOL deviation, review the literature on determining IOL power after refractive surgery, and introduce a formula for IOL calculation for use after refractive surgery for myopia. SETTING: Laser & Corneal Surgery Associates and Center for Ocular Tear Film Disorders, New York, New York, USA. METHODS: This retrospective noncomparative case series comprised 21 patients who had uneventful cataract extraction and IOL implantation after previous uneventful myopic refractive surgery. Six methods of IOL calculation were used: clinical history (IOL(HisK)), clinical history at the spectacle plane (IOL(HisKs)), vertex (IOL(vertex)), back calculated (IOL(BC)), calculation based on average keratometry (IOL(avgK)), and calculation based on flattest keratometry (IOL(flatK)). Each method result was compared to an "exact" IOL (IOL(exact)) that would have resulted in emmetropia and then compared to the pre-refractive surgery manifest refraction using linear regression. The paired t test was used to determine statistical significance. RESULTS: The IOL(HisKs) was the most accurate method for IOL calculations, with a mean deviation from emmetropia of -0.56 diopter +/-1.59 (D), followed by the IOL(BC) (+1.06 +/- 1.51 D), IOL(vertex) (+1.51 +/- 1.95 D), IOL(flatK) (-1.72 +/- 2.19 D), IOL(HisK) (-1.76 +/- 1.76 D), and IOL(avgK) (-2.32 +/- 2.36 D). There was no statistical difference between IOL(HisKs) and IOL(exact) in myopic eyes. The power of IOL(flatK) would be inaccurate by -(0.47x+0.85), where x is the pre refractive surgery myopic SE (SEQ(m)). Thus, without adjusting IOL(flatK), most patients would be left hyperopic. However, when IOL(flatK) is adjusted with this formula, it would not be statistically different from IOL(exact). CONCLUSIONS: For IOL power selection in previously myopic patients, a predictive formula to calculate IOL power based only on the pre-refractive surgery SEQ(m) and current flattest keratometry readings was not statistically different from IOL(exact). The IOL(HisKs), which was also not statistically different from IOL(exact), requires pre-refractive surgery keratometry readings that are often not available to the cataract surgeon. PMID- 15811747 TI - Evaluation of apparent ectasia of the posterior surface of the cornea after keratorefractive surgery. AB - PURPOSE: To calculate the apparent posterior corneal changes after keratorefractive surgery and reevaluate corneal ectasia displayed by Orbscan (Orbtek). SETTING: Department of Ophthalmology, Nara Medical University, Nara, Japan. METHODS: Postoperative:preoperative magnification ratio of the posterior surface of the cornea was calculated in a theoretical eye model. RESULTS: Assuming the preoperative corneal thickness is 600.00 microm, the preoperative refractive power of the anterior corneal surface is 48.0 diopters (D), the refractive power of the cornea is 1.376, the ablation diameter is 6.0 mm, the postoperative corneal thickness is 480.00 microm, the postoperative refractive power of the anterior corneal surface is 38.0 D, and the posterior surface of the cornea does not change postoperatively, the apparent image of the posterior surface of the cornea becomes 0.778% smaller postoperatively. If the posterior radius of curvature of the cornea is 6.2 mm, it becomes smaller by 48.24 microm. If this change directly affects the difference map, the posterior surface of the cornea moves forward by 48.24 microm. CONCLUSION: The results correspond to the amount of ectasia in previous reports. This artifact may explain the apparent ectasia detected by Orbscan. PMID- 15811748 TI - Optical performance of 3 intraocular lens designs in the presence of decentration. AB - PURPOSE: To study the theoretical optical performance of 3 intraocular lens (IOL) designs in the presence of IOL decentration. SETTING: Optics Center, Bausch & Lomb, Rochester, New York, USA. METHODS: A ray-tracing program was used to evaluate the effect of IOL decentration on the optical performance of 3 silicone IOLs (LI61U, Bausch & Lomb; Tecnis Z9000, Advanced Medical Optics; and a new aberration-free IOL [SofPort AO, Bausch & Lomb]) in an experimental model eye. The study was done using pupil diameters of 3.0 mm, 4.0 mm, and 5.0 mm and IOL decentrations of 0 mm, 0.25 mm, 0.50 mm, 0.75 mm, and 1.00 mm. The modulation transfer functions were computed and plotted. A Monte Carlo simulation analysis with 1000 trials with IOL decentration randomly varying for each pupil size was performed. RESULTS: Decentration of LI61U and Tecnis Z9000 IOLs led to asymmetrical higher-order aberrations that adversely affected the optical performance of the model eye; performance was not affected with the aberration free IOL because it lacks inherent spherical aberration. Optical performance with the aberration-free IOL was better than with the LI61U IOL as the former has less spherical aberration and did not introduce other aberrations when decentered. Performance with the aberration-free IOL was better than with the Tecnis Z9000 IOL for 3.0 mm, 4.0 mm, and 5.0 mm pupils when decentration exceeded 0.15 mm, 0.30 mm, and 0.38 mm, respectively. Performance with the LI61U IOL was better than with the Tecnis Z9000 IOL for 3.0 mm, 4.0 mm, and 5.0 mm pupils when decentration exceeded 0.3 mm, 0.5 mm, and 0.5 mm, respectively. Monte Carlo simulations showed the expected postoperative results of the LI61U IOL and aberration-free IOL would be repeatable and predictable, whereas the outcomes with the Tecnis Z9000 IOL would vary widely. CONCLUSIONS: The optical performance of the model eye was not affected by decentration of an aspheric IOL designed to have no inherent spherical aberration. With decentration, the performance with the new IOL was better than with a conventional spherical IOL and an aspheric IOL designed to offset the spherical aberration of an average cornea. PMID- 15811749 TI - Dioptric changes in eyes with reversed intraocular lenses. AB - PURPOSE: To theoretically calculate refractive changes in eyes with reversed intraocular lenses (IOLs). SETTING: Nara Medical University, Nara, Japan. METHODS: With the ray-tracing method, the refractive changes in a theoretical eye model with a reversed IOL in the capsular bag were calculated. Data for 3 models, anterior biconvex (MA60AC, Alcon), posterior biconvex (MA60BM, Alcon), and convex plano (UV2565T, Menicon), were provided by the manufacturer and used in the analysis. RESULTS: Using the assumption that the axial length varied from 22 to 27 mm, the anterior-biconvex IOLs elicited small refractive changes when reversed. The posterior-biconvex IOLs elicited a significant myopic shift (up to 0.90 diopter [D]) when reversed. The convex-plano IOLs elicited a significant hyperopic shift (up to 2.01 D) when reversed. CONCLUSIONS: Anterior-biconvex IOLs with similar anterior and posterior radii of curvature will not cause large diopter surprises when reversed. High-power posterior-convex IOLs will cause significant myopic diopter surprises when reversed. Both these changes shift toward myopic if the postoperative anterior chamber depth (ACD) becomes shallow. Medium- to high-power convex-plano IOLs will cause a large hyperopic shift when reversed. This hyperopic shift decreases if the postoperative ACD becomes shallower. PMID- 15811750 TI - Experimental observation of intraocular pressure changes during microkeratome suctioning in laser in situ keratomileusis. AB - PURPOSE: To observe changes in intraocular pressure (IOP), the manner in which the eye is suctioned, and the effect of the number of suction ports. SETTING: Department of Ophthalmology, Tokyo Dental College, Suidobashi Hospital, Tokyo, Japan. METHODS: Suction rings with single or dual ports were made of transparent acrylic acid resin to facilitate observation with a digital video camera. The IOP and the duration of the IOP increase (time to reach 90% of the maximum IOP) were measured in 6 porcine eyes with an intra-vein pressure-sensor catheter in the vitreous cavity. The IOP changes were recorded using a personal computer. RESULTS: Suctioning with single- and dual-port suction rings was similar. The IOP increases with single- and dual-port suction rings were similar (99.1 mm Hg +/- 6.1 [SD] and 99.0 +/- 6.5 mm Hg, respectively) (P=.987). The duration of the IOP increase was also similar (4.21 +/- 0.24 seconds and 4.12 +/- 0.33 seconds, respectively) (P=.190). CONCLUSIONS: The number of ports did not affect suctioning and changes in IOP. This technique is useful in developing the ideal shape and setting of the suction ring. PMID- 15811751 TI - Upregulation of alphavbeta6 integrin, a potent TGF-beta1 activator, and posterior capsule opacification. AB - PURPOSE: To identify the predominant activation pathway of transforming growth factor (TGF)-beta1 in the lens capsule, studying the spatial and temporal expression pattern of alphavbeta6 and thrombospondin-1. Other PCO-related proteins were also studied. SETTING: Departments of Ophthalmology and Optometrics and Clinical Pathology, Medical School, University of Vienna, Vienna, Austria. METHODS: The lens capsules of 12 human donor eyes were cultivated in a protein free medium for up to 28 days (cultivated lens capsules [CLCs]) after lens extraction. Ten intact lenses (ILs) served as the control group and were also cultured. During the culture period, cell dynamics were observed by phase contrast microscopy. Proteins were detected by double immunofluorescence on frozen sections. RESULTS: In ILs, alphavbeta6 was absent but 91.6% of the CLCs showed extensive staining. Remnant lens epithelial cells (LECs) expressed alphavbeta6 immediately after lens extraction. The alphavbeta6 was detected throughout the culture period in all regions of the capsule. Thrombospondin-1 was absent in ILs and CLCs, suggesting that this protein is not significant in TGF beta1 activation in the lens. Transforming growth factor-beta1 was abundantly expressed in all ILs and CLCs, slightly decreasing during intensive LEC proliferation and migration. The TGF-beta receptor II (RII) was expressed equally in all specimens, decreasing with culture time. Nonresident extracellular matrix proteins and alpha-smooth muscle actin were partially detected in CLCs but not in ILs. Latent TGF-beta binding protein 1 and collagen III were absent in all specimens. All cells found in the cultures expressed vimentin and alphaB crystallin (LEC markers). CONCLUSION: Alphavbeta6 is the main activator of TGF beta1 in the lens capsule and represents a new target for PCO prevention. PMID- 15811752 TI - Biocompatibility of poloxamer hydrogel as an injectable intraocular lens: a pilot study. AB - PURPOSE: To induce irreversible gelation of poloxamer, a thermosensitive polymer hydrogel, by using a photoinitiator and ultraviolet (UV) irradiation and to verify the biocompatibility and use of poloxamer as an injectable intraocular lens (IOL) material. SETTING: Department of Ophthalmology, Seoul National University College of Medicine, Seoul Artificial Eye Center, Clinical Research Institute, Seoul National University Hospital, Seoul, Korea. METHODS: In 10 rabbits, endocapsular phacoemulsification was performed and a poloxamer photoinitiator mixture was injected into the capsular bag through a small capsulorhexis site. In 1 eye, the capsulorhexis site was closed with a small plug and the entire eye was irradiated with UV light for 5 minutes. Postoperatively, poloxamer transparency and effect on the conjunctiva, cornea, iris, vitreous, and retina were observed. RESULTS: A mixture comprising 25% poloxamer and 0.01% photoinitiator produced a poloxamer that remained transparent in the lens capsule for up to 6 months. No inflammatory response or toxicity was observed in the conjunctiva, cornea, iris, vitreous, or retina. CONCLUSION: Poloxamer is a potentially suitable material for an injectable IOL. Further study is needed. PMID- 15811753 TI - Cellular death mediated by nuclear factor kappa B (NF-kappaB) translocation in cultured human lens epithelial cells after ultraviolet-B irradiation. AB - PURPOSE: To determine the role of nuclear factor kappa B (NF-kappaB) in the death of lens epithelial cells (LECs) after ultraviolet (UV) irradiation. SETTING: Department of Ophthalmology, Ilsan Paik Hospital, Inje University, Korea. METHODS: Cultures of simian virus 40 transfected human LECs (HLE B-3 cells) were were irradiated with a UVB source (312 nm) located 10 cm from the bottom of the slides for 1, 2, 3, or 4 minutes. Cytotoxicity was evaluated using a 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide method. Translocation of NF-kappaB was examined by immunocytochemistry using anti-NF-kappaB p65 antibody and electrophoretic mobility shift assay (EMSA). Sulfasalazine, a specific NF kappaB inhibitor, was used to confirm the role of NF-kappaB by pretreating samples for 30 minutes before UV irradiation, after which cytotoxicity and NF kappaB translocation were evaluated. RESULTS: When HLE B-3 cells were irradiated with UVB, translocation of NF-kappaB was observed with immunocytochemistry. These translocations peaked during EMSA 6 hours after UV irradiation. In HLE B-3 cells pretreated with sulfasalazine, the translocation of NF-kappaB was blocked. Cellular death after UV irradiation was also markedly reduced by sulfasalazine pretreatment. Ultraviolet irradiation can translocate NF-kappaB, and sulfasalazine is a useful blocking agent in this pathway. In this experimental model, sulfasalazine prevented cellular death after UV irradiation. CONCLUSION: The findings suggest that NF-kappaB plays an important role in cellular death after UV irradiation. PMID- 15811754 TI - Severe anaphylactic reaction after intracameral antibiotic administration during cataract surgery. AB - We report a severe anaphylactic reaction that occurred about 5 minutes after 1.0 mg of cefuroxime was injected into the anterior chamber after routine phacoemulsification and intraocular lens implantation. The patient was known to be allergic to penicillin. Immediate action was taken, and the patient recovered well. The presence of staff trained in resuscitation was essential in this case and raises questions about the trend to perform routine topical anesthesia cataract surgery without an anesthesiologist in the operating room. PMID- 15811755 TI - Recurrent Enterococcus faecalis endophthalmitis after phacoemulsification. AB - We present the first reported case of late recurrence of postoperative Enterococcus faecalis endophthalmitis after cataract surgery in a 45-year-old diabetic man. Culture-positive E faecalis endophthalmitis was diagnosed 2 days after uneventful phacoemulsification. Early vitrectomy with intravitreal and subconjunctival vancomycin and amikacin and topical vancomycin resulted in apparently complete clinical resolution after 4 months and a best corrected visual acuity of 20/25. Recurrent endophthalmitis with hypopyon occurred 7 months postoperatively and resolved with intravitreal vancomycin and topical prednisolone acetate 1%. However, the patient had a similar relapse at 9 months that resulted in deterioration of visual acuity to no light perception despite a repeat vitrectomy, intraocular lens explantation, capsular bag removal, and intravitreal antibiotics. The late recurrences could have been the result of persistent sequestration of the organism in the capsular bag. PMID- 15811756 TI - Retained ophthalmic viscosurgical device material in the capsular bag 6 months after phacoemulsification. AB - We present a 63-year-old man who developed low-grade endophthalmitis 6 months after phacoemulsification and implantation of a foldable posterior chamber intraocular lens (IOL). The capsular bag was distended, but shallowing of the anterior chamber was not observed. Chemical analysis of the clear fluid aspirated from the capsular bag behind the IOL revealed the presence of an ophthalmic viscosurgical device. PMID- 15811757 TI - Late postoperative capsular block syndrome: entrapment of liquefied after cataract by capsular bend. AB - We report a case of capsular bend-related entrapment of liquefied after-cataract that resulted in late postoperative capsular block syndrome in a 56-year-old man. Slitlamp examination showed a capsular bend formation at the square edge of the optic. Superiorly, leakage in the capsular bend resulted in fluid extending into Soemmering's ring, although communication with the anterior chamber was limited by the capsular bend. Fluid was seen between the lens and posterior capsule. PMID- 15811758 TI - Late-onset repetitive traumatic flap folds and partial dehiscence of flap edge after laser in situ keratomileusis. AB - A 25-year-old woman had traumatic flap folds and partial dehiscence of the flap edge in the right eye 5 and 30 months after laser in situ keratomileusis. The period from injury to treatment was 4 hours and 9 days, respectively. With the first injury, the flap was lifted and stretched with moistened sponges to clear the folds. With the second injury, the folds were hard so the flap was lifted and sutured to stretch the folds. The patient recovered visual acuity within 2 weeks. PMID- 15811759 TI - Surface and Orbscan II slit-scanning elevation topography in circumscribed posterior keratoconus. AB - We used an Orbscan II topography system (Bausch & Lomb) to study anterior and posterior surface abnormalities, keratometry, and topographic pachymetry in a patient with circumscribed posterior keratoconus. This system clearly showed a marked localized paracentral annular elevation in the posterior corneal surface that corresponded to an abrupt decrease in thickness and a slightly localized anterior surface bulge in the anterior float. PMID- 15811760 TI - Shark fin: a new sign of thermal injury. AB - Three cases are described in which a curvilinear lucency in the corneal stroma signified a surgical thermal injury. The "shark fin" appearance represents focal shrinkage of tissue within the incisional tunnel. PMID- 15811761 TI - Nutritional status of Khasi schoolgirls in Meghalaya. AB - OBJECTIVE: The nutritional status of 222 Khasi girls within age groups 4 to 6 y, 7 to 9 y, and 10 to 12 y was studied. METHODS: Personal interviews using questionnaires and 24-h dietary recall were used. Weights and heights were recorded, body mass index was calculated, and children were classified as normal or malnourished using Z scores and growth charts from the Centers for Disease Control and Prevention. Based on weights and heights, children were classified as normal or undernourished according to classifications by Gomez, Waterlow, and the National Center for Health Statistics. RESULTS: Heights of girls ages 7 to 9 y and 10 to 12 y and weights of all girls were significantly (P < 0.05) lower than the reference values for height (126.4 and 142.7 m) and weight (19, 26.9, and 31.5 kg), respectively. The lowest Z scores (-1.5 to -1.0) and percentiles (10th) for body mass index were seen in 12-y-old girls. Most girls ages 7 to 9 y had grade I malnutrition. Girls ages 10 to 12 y had the highest incidence of moderate malnutrition. Severe stunting was observed in all groups. Average energy consumption was significantly (P < 0.05) lower than the recommended dietary allowance in all the age groups. Consumption of protein by children ages 7 to 9 y and 10 to 12 y was also significantly lower than the recommended dietary allowance. Consumption of calcium, iron, and carotene in children 10 to 12 y old was significantly (P < 0.05) lower than the recommended dietary allowance. CONCLUSIONS: Sociologic community factors are required to facilitate implementation of a nutritional package and availability of key nutrients to ensure growth in children. PMID- 15811762 TI - An anti-stray stylet for subclavian catheterization by the infraclavicular approach. AB - OBJECTIVE: Misplacement of subclavian vein catheters has been reported. We assessed the rate of misplacement of a subclavian vein catheter with and without a novel stylet. METHODS: A prospective, randomized, controlled clinical trial was conducted between September 2001 and June 2003 in a university hospital. Two hundred sixteen adult patients were enrolled to receive subclavian vein catheterization under non-emergency conditions. Patients were randomly assigned to undergo right subclavian vein catheterization with the stylet (n = 109, stylet group) or without the stylet (n = 107, control group). The rate of subclavian vein catheter misplacement was determined and risk factors for failure and complications were analyzed. RESULTS: There was no incidence of catheter misplacement in the stylet group, but this occurred in 12 patients in the control group in whom the catheter was misplaced into the ipsilateral internal jugular vein (0% versus 11.2%, P = 0.0003). In multivariate analyses, use of the stylet (odds ratio = 0.062, 95% confidence interval = 0.008 to 0.495, P = 0.009) and a close to average body mass index (odds ratio = 0.73, 95% confidence interval = 0.54 to 0.98, P = 0.038) were associated with low risks of complications and failure, respectively. CONCLUSIONS: This stylet is useful for decreasing the incidence of misplacement of subclavian vein catheters. Body mass index is predictive of failed vein puncture. PMID- 15811763 TI - Sodium depletion and hemoconcentration: overlooked complications in patients with anorexia nervosa? AB - OBJECTIVE: This study evaluated sodium and fluid balance disturbances in anorexia nervosa. METHODS: Serum urea, creatinine, urea nitrogen/creatinine ratio, protein concentration, osmolality, electrolytes, hemoglobin, and hematocrit were evaluated on admission, after intravenous fluid supplementation, and at discharge in 14 patients who had anorexia nervosa and were admitted to a medical unit for severe malnutrition and/or medical complications. Diet history and physical signs of salt and water depletion were also evaluated on admission. The diagnosis of plasma volume depletion was considered confirmed if serum urea nitrogen/creatinine ratio was higher than 25. RESULTS: Mean body mass indexes were 12.9 +/- 2.4 kg/m2 on admission and 14.2 +/- 2.0 kg/m2 at discharge. Mean duration of hospitalization was 24.2 +/- 11.9 d. Plasma volume depletion was found on admission in nine patients (64%), and hyponatremia was associated with hypovolemia (hypovolemic hyponatremia) in three of them. Most of the commonly used indexes of hemoconcentration (hematocrit, hemoglobin, serum urea, and creatinine) were normal on admission but decreased significantly after intravenous fluid supplementation and were maintained at lower levels at discharge (P < 0.001, P < 0.0004, P < 0.009, and P < 0.001, respectively, compared with admission values). Hemoconcentration masked an underlying anemia (hemoglobin < 12 g/dL) in six patients. Only two of the nine patients with plasma volume depletion had a history of diuretic abuse, and the other seven patients reported long-term sodium restriction as a method to control body weight. CONCLUSIONS: Plasma volume depletion and hypovolemic hyponatremia are common in the most severely malnourished patients with anorexia nervosa. Clinicians should be aware that, because of malnutrition, the common indexes of hemoconcentration may be within normal range and that hemoconcentration may mask anemia. Further studies are needed to assess the role of neglected behaviors such as sodium and fluid restriction. PMID- 15811764 TI - Influence of heredity on dietary restraint, disinhibition, and perceived hunger in humans. AB - OBJECTIVE: Dietary restraint, disinhibition, and perceived hunger have been shown to affect food intake and body weight and are thought to be risk factors for eating disorders, but little is known about their origins. We investigated the influence of heredity, shared (familial) environment, and individual environment on dietary restraint disinhibition, perceived hunger and their relation to body size and food intake. METHODS: Scores on the Three Factor Eating Questionnaire and the Restraint Scale in addition to height, weight, body mass index, and 7-d diary reported nutrient intakes were obtained from 39 identical, 60 fraternal same-sex, and 50 fraternal opposite-sex adult twin pairs who were living independently. Linear structural modeling was applied to investigate the nature and degree of genetic and environmental influences. RESULTS: Analysis showed significant genetic and individual environmental, but not shared (familial) environmental, influences on cognitive restraint, perceived hunger, and Restraint Scale scores, with genes accounting for 44%, 24%, and 58% of the variance, respectively. In contrast, disinhibition was found to be significantly influenced by the shared (familial) environment, accounting for 40% of the variance. Further analysis showed that cognitive restraint and perceived hunger heritabilities could not be accounted for by significant heritabilities of body weight, height, or body mass index. In contrast, the heritability of Restraint Scale scores was found to be related to body size. Cognitive restraint was negatively correlated with nutrient intake, and differences in cognitive restraint were found to be related to differences in the body sizes of identical twin pairs. CONCLUSIONS: Dietary restraint appears to be another component in a package of genetically determined physiologic, sociocultural, and psychological processes that regulate energy balance, whereas dietary disinhibition may be the intermediary between upbringing and the development of overweight and/or eating disorders. PMID- 15811765 TI - Association between gestational night blindness and serum retinol in mother/newborn pairs in the city of Rio de Janeiro, Brazil. AB - OBJECTIVE: Gestational night blindness (XN) is associated with increased risk of reproductive morbidity and mortality. This study investigated the prevalence of gestational XN among postpartum women treated in a public maternity hospital in the city of Rio de Janeiro, Brazil and evaluated its association with maternal and neonatal (cord blood) serum retinol concentrations. METHODS: XN was evaluated retrospectively, using an interview according to guidelines of the World Health Organization, in 222 postpartum women (< or = 6 h after delivery) after singleton births who had low obstetric risk. Serum retinol concentrations were measured according to the modified Bessey method, with a cutoff point lower than 1.05 micromol/L for inadequate serum retinol concentration. RESULTS: Prevalence of gestational XN was 18%, and inadequate maternal and cord blood serum retinol concentrations were found in 24.4% and 45.5% of samples, respectively. The results associated gestational XN with inadequate maternal serum retinol concentration (P = 0.000), and an association was observed between maternal and neonatal serum retinol concentrations (P = 0.000). A poor association was observed between maternal XN and serum levels of retinol in newborn children (P = 0.06). CONCLUSIONS: The results suggest that prevalence of gestational XN and inadequate serum retinol concentration among postpartum women and newborns is a concern, calling attention to the need for studies in other parts of Brazil. In addition, the risk of inadequate serum retinol in newborns was significantly higher among infants of postpartum women with serum retinol levels below 1.05 micromol/L. Gestational XN was associated with inadequate levels of maternal serum retinol, and the results suggest a poor relation between maternal XN and vitamin A nutritional status of newborns. PMID- 15811766 TI - Does dietary recall adequately assess sodium, potassium, and calcium intake in hypertensive patients? AB - OBJECTIVE: A diet low in sodium, high in potassium, and high in calcium is recommended to lower blood pressure. However, compliance with this diet is poor, probably because of dietary intake underestimation. Therefore, we compared electrolyte intake as estimated from dietary recall with a 24-h urinary excretion. METHODS: Thirty-six patients (26 men and 10 women) with a mean age of 46 +/- 8 y participated in the study. All participants had essential hypertension and were on no drug therapy (n = 20) or non-diuretic monotherapy (n = 16). Patients were instructed to consume a low-sodium (50 mmol/d), high-potassium (supplementation with 30 to 60 mmol/d), and high-calcium (1000 mg/d) diet. Compliance with the diet was assessed at baseline and then 1, 2, and 3 mo after starting the diet. Sodium, potassium, and calcium intakes were carefully estimated from patients' dietary recall and 24-h urinary collection. RESULTS: Estimated sodium intake significantly correlated with 24-h urinary excretion (R = 0.43 P < 0.001). However, estimated sodium intake was lower than urinary sodium excretion by 34% at baseline and by 47% after 3 mo of dieting (P < 0.05). Estimated potassium intake correlated with 24-h urinary excretion. Estimated calcium intake significantly increased from 933 +/- 83 mg/d to 1029 +/- 171 mg/d (P < 0.05). Calcium intake derived from patients' recall far exceeded and only slightly correlated with 24-h urinary excretion (R = 0.23, P < 0.01). CONCLUSIONS: Patients tend to underestimate their sodium intake by 30% to 50%; therefore, urinary sodium excretion is more accurate to assess sodium intake. Thus, 24-h urinary sodium excretion should be used in clinical practice and in clinical trials, especially when dietary non-compliance is suspected. PMID- 15811767 TI - Lipid composition in human breast milk from Granada (Spain): changes during lactation. AB - OBJECTIVE: To determine possible differences of composition in the course of lactation, phospholipid (PL) classes (phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin) and fatty acid composition of PL and triacylglycerol (TGs) fractions of milk fat were analyzed in 66 samples from mothers from Granada (Spain) who gave birth to full-term infants. Analyses included colostrum, transitional milk, and mature milk. METHODS: After milk fat extraction, PLs and TGs were separated by thin layer chromatography and fatty acids of each fraction were converted into their methyl esters, which were analyzed by gas chromatography. PL classes were determined by high-performance liquid chromatography using an evaporative light scattering detector. RESULTS: Mature human milk showed a lower content (P = 0.020) of PLs than did the other milks. Percentage of sphingomyelin was constant for all stages of lactation, whereas the percentage of phosphatidylcholine in mature milk was significantly lower (P < 0.05) than in colostrum and transitional milk. TGs in mature human milk contained lower percentages (P < 0.001) of arachidonic acid, docosahexaenoic acid, and nervonic acid. Docosahexaenoic acid and nervonic acid also showed a significant decrease (P < 0.001) in total PLs from colostrum and mature milk. CONCLUSIONS: The composition of PL classes and fatty acids in PLs and TGs in milk of mothers in Granada (Southern Europe) is different from that in milk from mothers in other parts of the world. In addition, the ratio of long-chain polyunsaturated fatty acids delivered in the form of PLs to long-chain polyunsaturated fatty acids delivered in the form of triacylglycerols diminishes as lactation proceeds. PMID- 15811768 TI - Early infant diet and risk of type 1 diabetes mellitus in Belgrade children. AB - OBJECTIVES: This study investigated whether an infant diet is associated with the development of type 1 diabetes. METHODS: A case-control study was conducted in Belgrade from 1994 to 1997. A total of 105 patients with recent onset diabetes (< or = 16 y old) were compared with 210 controls chosen among children with skin disease (first control group). Cases and controls were individually matched by age (+/-1 y), sex, and place of residence. Eighty-six children with diabetes were also compared with their siblings (second control group). RESULTS: According to univariate logistic regression analysis, when cases were compared with the first control group, the risk of type 1 diabetes was greater for children who were breast fed less than 4 mo (odds ratio = 2.09, 95% confidence interval = 1.30 to 3.36) and who received cow's milk at younger than 5 mo (odds ratio = 3.39, 95% confidence interval = 2.04 to 5.66). According to univariate analysis, when cases were compared with their relatives, only early introduction of supplementary milk was associated with a higher risk for diabetes (odds ratio = 5.75, 95% confidence interval = 2.91 to 11.36). After adjusting for different confounding variables, infant diet was not independently associated with diabetes. CONCLUSIONS: The results obtained do not support the hypothesis that infant diet is related to the occurrence of type 1 diabetes. PMID- 15811769 TI - Usefulness of granular BCAA after hepatectomy for liver cancer complicated with liver cirrhosis. AB - OBJECTIVE: Nutritional disturbances such as ascites and hypoalbuminemia frequently arise after hepatectomy for liver cancer with liver cirrhosis. We examined the possibility of maintaining a favorable state of nutrition by outpatient administration of branched-chain amino acid (BCAA) granules. METHODS: Forty-three patients who had gross liver cirrhosis complicated by liver cancer and underwent surgery up to May 2002 were given BCAA granules (n = 21, BCAA group) or no granules (n = 22, control group). RESULTS: 1) Background details such as age, sex, surgical technique, blood loss, and duration of surgery showed no significant differences. 2) Among objective findings, improvement of ascites and edema tended to occur sooner in the BCAA group, but without a significant difference. 3) Although serum albumin recovered its preoperative value 9 mo after surgery in the control group, only 6 mo was required for recovery in the BCAA group. Total protein showed similar changes, but neither group showed any difference in changes of aspartate aminotransferase, alanine transferase, or platelets. 4) One year postoperatively, the change from the preoperative indocyanine green retention rate at 15 min after intravenous administration tended to be worse in the control group, but not significantly so. 5) In the BCAA group, hyaluronic acid and type IV collagen 7S improved significantly sooner than in the control group. CONCLUSIONS: BCAA supplementation after hepatectomy promotes rapid improvement in protein metabolism and inhibits progression to liver cirrhosis. Administration of BCAA after hepatectomy is considered beneficial to a patient's nutritional state. PMID- 15811770 TI - State of malnutrition in Cuban hospitals. AB - OBJECTIVE: We assessed the current state of undernutrition as observed in 1905 patients hospitalized in 12 Cuban health care institutions, as part of a Latin American, multinational survey similar in design and goals. METHODS: We surveyed 1905 randomly selected patients from 12 Cuban hospitals in a two-phase study. Patients' clinical charts were audited in phase 1, the Subjective Global Assessment was used to assess patients' nutritional status in phase 2. The study was locally conducted by a properly trained team. RESULTS: The frequency of undernutrition in Cuban hospitals was 41.2% (95% confidence interval = 38.9 to 43.4), and 11.1% of patients were considered severely undernourished. Statistically significant (P < 0.05) univariate relations were identified between undernutrition and patient's age and sex. Nutritional status was a dependent of the patient's instruction level (P < 0.05). Patients' nutritional status was statistically associated with the presence of cancer and infection. Undernutrition was highly prevalent among cancer patients, no matter the stage of medical or surgical treatment. Undernutrition became extremely frequent after surgical treatment in non-cancer patients. High nutritional risks hospital services/specialties were identified: geriatrics (56.3%), critical care (54.8%), nephrology (54.3%), internal medicine (48.6%), gastroenterology (46.5%), and cardiovascular surgery (44.8%). Malnutrition rates increased progressively with prolonged length of stay. CONCLUSION: A high malnutrition rate was observed among participating hospitals. The design and inception of policies that foster intervention programs focusing on early identification of hospital malnutrition and its timely management is suggested to decrease its deleterious effects on outcomes of health care in the participating hospitals. PMID- 15811771 TI - Evaluation of Mini-Nutritional Assessment for Japanese frail elderly. AB - OBJECTIVE: We evaluated the Mini-Nutritional Assessment (MNA) test and the short form MNA as screening tools for malnutrition in the Japanese elderly population. METHODS: A cross-sectional study of 226 elderly Japanese patients (78.6 +/- 0.5 y of age, mean +/- standard deviation; 67 men and 159 women) in various settings was carried out. Nutritional assessment included MNA, anthropometric measurements, and biochemical markers. RESULTS: According to the original cutoff point of the full MNA, 19.9% of those assessed were malnourished, 58.0% were at risk of malnutrition, and 22.1% were well nourished. Significant correlations were found between full MNA scores and age (r = -0.14), body mass index (r = 0.59), serum albumin (r = 0.60), total cholesterol (r = 0.36), midarm circumference (r = 0.50), and triceps skinfold (r = 0.37). The sensitivity and specificity of the full MNA score (< 17) for hypoalbuminemia were 0.810 and 0.860, respectively. With a cutoff point lower than 18, sensitivity and specificity hypoalbuminemia were 0.857 and 0.815, respectively. Using a short form MNA score 12 and higher as normal, its sensitivity and specificity for predicting undernutrition were 0.859 and 0.840, respectively. CONCLUSIONS: The full and short forms of the MNA were useful tools to identify elderly Japanese patients with malnutrition or risk of malnutrition. However, the full MNA cutoff point for malnutrition should be modulated for this population. PMID- 15811772 TI - Protective effect of dietary azuki bean (Vigna angularis) seed coats against renal interstitial fibrosis of rats induced by cisplatin. AB - OBJECTIVE: We investigated the effects of azuki bean (Vigna angularis) seed coats (ABSCs), which mainly contain proanthocyanidins and dietary fibers, on the infiltration of macrophages and the progression of renal interstitial fibrosis induced by cisplatin (CDDP). METHODS: Male rats were divided into two groups: controls received intraperitoneal injections of saline and the other rats were injected intraperitoneally with 3 mg of CDDP per kilogram once a week for 5 wk. The CDDP-injected animals received one of four diets: 1) control diet (commercial diet only), 2) 0.5% red ABSC (RABSC) diet, 3) 2.0% RABSC diet, and 4) 2.0% white ABSC (WABSC) diet. The saline-injected animals were given the commercial diet. Five weeks after the final CDDP injection, macrophage kinetics and interstitial fibrotic areas were examined. RESULTS: The content of polyphenols in the RABSC (76.3 g/kg of plant material) was higher than that in the WABSC (18.1 g/kg). Proanthocyanidins were detected in the RABSC (20.4 g/kg) but not in the WABSC. Histologically, the fibrotic areas consisting of fibroblastic cells and mononuclear cells developed around the dilated or atrophic tubules in the corticomedullary junction in CDDP-treated rat kidney, whereas the extent and magnitude of damage were less in the WABSC- and RABSC-treated rats. In immunohistochemical analysis, ED1-positive macrophages in CDDP-treated rats showed a significant increase in number compared with the control. The number of macrophages in CDDP plus WABSC or RABSC groups was significantly smaller than that in CDDP-treated rats. In addition, the number of macrophages in the RABSC group was significantly smaller than in the WABSC group, indicating that ABSC, especially RABSC, prevented macrophages from infiltrating into areas of interstitial fibrosis. CONCLUSIONS: These results suggest that ABSC, especially RABSC, suppress the increase of infiltrating macrophages in the damaged kidney and may lead to amelioration of interstitial fibrosis. Based on the composition of ABSC, molecules such as proanthocyanidins and/or dietary fibers may be associated with the amelioration of renal damage. PMID- 15811773 TI - Effects of conjugated linoleic acid on liver composition and fatty acid oxidation are isomer-dependent in hamster. AB - OBJECTIVE: The present work was designed to study the effects of the two main isomers of conjugated linoleic acid (CLA), cis-9,trans-11 and trans-10,cis-12, on liver composition and hepatic fatty acid oxidation in hamsters. METHODS: Animals were divided into three groups that were fed atherogenic diets supplemented with 0.5% linoleic acid, cis-9,trans-11 CLA, or trans-10,cis-12 CLA for 6 wk. Liver lipids, protein, water and DNA contents, and histologic structure were analyzed. Hepatic carnitine palmitoyltransferase-I and acyl coenzyme A oxidase activities were assessed. Triacylglycerol concentration, and aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and alkaline phosphatase activities were evaluated in serum. CLA isomer contents were analyzed by gas chromatography in hepatic triacylglycerols. Peroxisome proliferator-activated receptor-alpha mRNA was determined by reverse transcriptase polymerase chain reaction. RESULTS: Trans-10,cis-12 CLA led to significantly greater weight, lower levels of triacylglycerol, cholesterol, and phospholipid, and larger total cell number in liver. Carnitine palmitoyltransferase-I and acyl coenzyme A oxidase activities were significantly increased by this isomer. No changes were induced by cis-9,trans-11 CLA. Trans-10,cis-12 CLA was recovered in significantly lower proportions than cis-9,trans-11 in liver triacylglycerols. Histopathologic analysis showed no abnormalities. No significant differences in serum aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and alkaline phosphatase activities or in hepatic mRNA peroxisome proliferator activated receptor-alpha expression were found among the three experimental groups. CONCLUSIONS: These results suggest that the addition of 0.5% of these CLA isomers to the diet do not induce toxic effects in liver after 6 wk of feeding. Intake of trans-10,cis-12 isomer but not of cis-9,trans-11 CLA increases liver fatty acid oxidation. This effect leads to decreased hepatic and serum triacylglycerols. PMID- 15811774 TI - Recovery from iron deficiency in rats by the intake of recombinant yeast producing human H-ferritin. AB - OBJECTIVE: This study examined whether iron accumulated in ferritin-producing recombinant microbes is bioavailable to rats with iron deficiency. METHODS: Rats induced with iron deficiency were treated with iron preparations of ferrous ammonium sulfate, horse spleen ferritin, control yeast, and ferritin-producing recombinant yeast for 14 d. The bioavailability of iron was examined by measuring hemoglobin concentration, hematocrit value, and tissue iron stores. Differences between dietary groups were determined by one-way analysis of variance, and P < 0.05 was considered statistically significant. RESULTS: Based on hemoglobin concentration and hematocrit value, iron in ferrous ammonium sulfate, horse spleen ferritin, and ferritin-producing yeast were bioavailable to rats and cured iron deficiency. The efficacy of ferritin and ferritin-producing yeast was also confirmed in establishing tissue iron stores after induction of iron deficiency. CONCLUSIONS: The iron sources of ferritin and ferritin-producing yeast were as effective in recovery from iron deficiency as the iron compounds of ferric citrate and ferrous ammonium sulfate. The results suggest that the iron stored in the ferritin of recombinant yeast is bioavailable, and that recombinant yeast may contribute widely as a source of iron to resolve the global problem of iron deficiency. PMID- 15811775 TI - Ethyl alpha-D-glucoside was absorbed in small intestine and excreted in urine as intact form. AB - OBJECTIVE: Ethyl alpha-D-glucoside (alpha-EG) is a peculiar component in sake. We investigated how alpha-EG was absorbed, hydrolyzed, and excreted in urine when it was ingested orally by rats. METHODS: Hydrolyzing activity for alpha-EG was determined by incubating it with crude enzyme solutions prepared from several rat organs, and absorption activity for alpha-EG was determined by incubating rat small intestinal everted sac in sodium or potassium Krebs-Ringer buffer that contained alpha-EG. alpha-EG solution was fed to rats, and urine volume and plasma alpha-EG, glucose and insulin and urinary alpha-EG were determined. RESULTS: alpha-EG was hydrolyzed by crude enzyme solutions prepared from rat small intestinal mucosa and kidney, and these hydrolyzing activities were lower than those for maltose. alpha-EG absorbed into everted rat intestinal sacs in potassium Krebs-Ringer buffer reduced almost completely compared with that in sodium Krebs-Ringer buffer. When alpha-EG was ingested orally by rats, it was absorbed into the bloodstream and more than 60% was excreted in urine, and urine volume increased. CONCLUSIONS: In rats, alpha-EG was absorbed in small intestine and excreted intact in urine without affecting blood glucose and insulin and thus was a diuretic, insulin-independent, and low-nutritive glucoside that could be safely applicable to food. PMID- 15811776 TI - Differential effects of organosulfur compounds from garlic oil on nitric oxide and prostaglandin E2 in stimulated macrophages. AB - OBJECTIVE: We investigated the inhibition of nitric oxide (NO) and prostaglandin E2 (PGE2) production by the garlic oil derivatives, diallyl sulfide (DAS), diallyl disulfide (DADS), and allyl methyl sulfide (AMS), in lipopolysaccharide (LPS)-activated RAW 264.7 cells. METHODS: Cells were treated with LPS (330 ng/mL) and various concentrations of DAS, DADS, and AMS. NO and PGE2 released into the medium and expressions of inducible NO synthase and cyclooxygenase-2 protein were measured. RESULTS: All three compounds suppressed stimulated NO production, among which AMS exhibited the least inhibition. Western blot analysis showed that DAS and DADS, but not AMS, inhibited the corresponding inducible NO synthase expression. An in vitro study showed that all three compounds possess NO clearance activity, and that DADS and AMS were more effective than DAS. On the contrary, only DAS inhibited activated PGE2 production and cyclooxygenase-2 protein expression. CONCLUSIONS: The garlic derivatives, DAS, DADS, and AMS, differentially regulated the production of NO and PGE2 in stimulated macrophages. DAS decreased stimulated NO and PGE2 production by inhibiting inducible NO synthase and cyclooxygenase-2 expressions, and its enzyme inhibiting and NO clearance activity may also partly contribute to the suppression of NO. DADS inhibited activated NO production by decreasing inducible NO synthase expression and by directly clearing NO, whereas AMS suppressed NO mainly through its direct NO clearance activity. Further, neither DADS nor AMS showed any inhibitory effect on stimulated PGE2 production. PMID- 15811777 TI - Effect of starvation on hepatic acyl-CoA synthetase, carnitine palmitoyltransferase-I, and acetyl-CoA carboxylase mRNA levels in rats. AB - OBJECTIVES: This study investigated the effect of starvation on mRNA levels of hepatic acyl coenzyme A synthetase (ACS), carnitine palmitoyltransferase-I (CPT I), and acetyl coenzyme A carboxylase (ACC) and on serum concentrations of leptin, insulin, and glucose in male Sprague-Dawley rats. METHODS: Rats were fed an AIN-76 diet for 5 wk and then assigned to a normal group (NG) and a starvation group (SG). The SG was starved for 48 h and the NG was fasted for 12 h before being killed. Serum and hepatic lipids and serum levels of leptin, insulin, and glucose were determined. Expressions of ACS, CPT-1, and ACC mRNA were assessed in liver. RESULTS: Serum concentrations of triacylglycerol and high-density lipoprotein cholesterol in the SG were lower than those in the NG. Serum concentrations of low-density lipoprotein cholesterol in the SG were significantly higher than in the NG. Hepatic concentrations of total lipid in the SG were significantly higher than those in the NG, and triacylglycerol concentrations in the SG were significantly lower than those in the NG. Serum concentrations of leptin and glucose in the SG were significantly lower than those in the NG. The ratio of abdominal fat to total body weight in the SG was lower than that in the NG. Hepatic ACS and CPT-I mRNA levels in the SG were significantly higher than those in the NG, but hepatic ACC mRNA levels were lower in the SG than in the NG. CONCLUSIONS: We demonstrated that starvation increases hepatic levels of ACS and CPT-I and decreases transcription levels of ACC, implicating increases in fatty acid oxidation. This research demonstrates a coordinated regulation of ACS, CPT-I, and ACC mRNA levels and serves to enhance our understanding of the molecular mechanisms underlying fatty acid metabolism during starvation. PMID- 15811778 TI - Differential regulation of hepatic gene expression by starvation versus refeeding following a high-sucrose or high-fat diet. AB - OBJECTIVES: The objective of this work was to determine the effects of starvation versus refeeding following a high-sucrose diet (HS) or high-fat diet (HF) on fatty acid metabolism in mice. METHODS: The mice were fed an AIN-76 control diet (CD), a modified HS, or an HF. The three dietary groups were subdivided into three groups each: those fed experimental diets for 12 wk, mice starved for 48 h after 12 wk on an experimental diet, and those with the same starvation treatment but with 72 h of refeeding after starvation, respectively. RESULTS: Serum total cholesterol levels of CD and HF groups decreased and then increased under starvation and refeeding states, respectively. Refeeding HS and HF increased serum levels of low-density lipoprotein (LDL) cholesterol compared with refeeding of the CD group. Starvation significantly increased hepatic levels of total cholesterol in the HS and HF groups compared with the CD group. Hepatic acyl coenzyme A (CoA) synthetase (ACS) levels in the CD and HS groups but not the HF group increased and then decreased under starved and refed states, respectively; an opposite regulation was observed in the HF group. Levels of hepatic acetyl-CoA carboxylase (ACC) in the HS and HF groups were significantly increased by refeeding. Hepatic levels of carnitine palmitoyltransferase-I mRNA were significantly enhanced by starvation and refeeding in the HS group but decreased in CD and then increased in the HF group. CONCLUSIONS: Changes in dietary energy nutrients, fasting, and refeeding affect hepatic ACS, CPT-I, and ACC mRNA expression, and these results will serve to enhance our understanding of the molecular mechanisms underlying regulation of fatty acid metabolism. PMID- 15811779 TI - Information extraction and summarization from medical documents. PMID- 15811780 TI - Automatic processing of multilingual medical terminology: applications to thesaurus enrichment and cross-language information retrieval. AB - OBJECTIVES: We present in this article experiments on multi-language information extraction and access in the medical domain. For such applications, multilingual terminology plays a crucial role when working on specialized languages and specific domains. MATERIAL AND METHODS: We propose firstly a method for enriching multilingual thesauri which extracts new terms from parallel corpora, and secondly, a new approach for bilingual lexicon extraction from comparable corpora, which uses a bilingual thesaurus as a pivot. We illustrate their use in multi-language information retrieval (English/German) in the medical domains. RESULTS: Our experiments show that these automatically extracted bilingual lexicons are accurate enough (85% precision for term extraction) for semi automatically enriching mono- or bi-lingual thesauri such as the universal medical language system, and that their use in cross-language information retrieval significantly improves the retrieval performance (from 22 to 40% average precision) and clearly outperforms existing bilingual lexicon resources (both general lexicons and specialized ones). CONCLUSION: We show in this paper first that bilingual lexicon extraction from parallel corpora in the medical domain could lead to accurate, specialized lexicons, which can be used to help enrich existing thesauri and second that bilingual lexicons extracted from comparable corpora outperform general bilingual resources for cross-language information retrieval. PMID- 15811781 TI - Bio-medical entity extraction using support vector machines. AB - OBJECTIVE: Support vector machines (SVMs) have achieved state-of-the-art performance in several classification tasks. In this article we apply them to the identification and semantic annotation of scientific and technical terminology in the domain of molecular biology. This illustrates the extensibility of the traditional named entity task to special domains with large-scale terminologies such as those in medicine and related disciplines. METHODS AND MATERIALS: The foundation for the model is a sample of text annotated by a domain expert according to an ontology of concepts, properties and relations. The model then learns to annotate unseen terms in new texts and contexts. The results can be used for a variety of intelligent language processing applications. We illustrate SVMs capabilities using a sample of 100 journal abstracts texts taken from the {human, blood cell, transcription factor} domain of MEDLINE. RESULTS: Approximately 3400 terms are annotated and the model performs at about 74% F score on cross-validation tests. A detailed analysis based on empirical evidence shows the contribution of various feature sets to performance. CONCLUSION: Our experiments indicate a relationship between feature window size and the amount of training data and that a combination of surface words, orthographic features and head noun features achieve the best performance among the feature sets tested. PMID- 15811782 TI - Comparative experiments on learning information extractors for proteins and their interactions. AB - OBJECTIVE: Automatically extracting information from biomedical text holds the promise of easily consolidating large amounts of biological knowledge in computer accessible form. This strategy is particularly attractive for extracting data relevant to genes of the human genome from the 11 million abstracts in Medline. However, extraction efforts have been frustrated by the lack of conventions for describing human genes and proteins. We have developed and evaluated a variety of learned information extraction systems for identifying human protein names in Medline abstracts and subsequently extracting information on interactions between the proteins. METHODS AND MATERIAL: We used a variety of machine learning methods to automatically develop information extraction systems for extracting information on gene/protein name, function and interactions from Medline abstracts. We present cross-validated results on identifying human proteins and their interactions by training and testing on a set of approximately 1000 manually-annotated Medline abstracts that discuss human genes/proteins. RESULTS: We demonstrate that machine learning approaches using support vector machines and maximum entropy are able to identify human proteins with higher accuracy than several previous approaches. We also demonstrate that various rule induction methods are able to identify protein interactions with higher precision than manually-developed rules. CONCLUSION: Our results show that it is promising to use machine learning to automatically build systems for extracting information from biomedical text. The results also give a broad picture of the relative strengths of a wide variety of methods when tested on a reasonably large human annotated corpus. PMID- 15811783 TI - Summarization from medical documents: a survey. AB - OBJECTIVE: The aim of this paper is to survey the recent work in medical documents summarization. BACKGROUND: During the last decade, documents summarization got increasing attention by the AI research community. More recently it also attracted the interest of the medical research community as well, due to the enormous growth of information that is available to the physicians and researchers in medicine, through the large and growing number of published journals, conference proceedings, medical sites and portals on the World Wide Web, electronic medical records, etc. METHODOLOGY: This survey gives first a general background on documents summarization, presenting the factors that summarization depends upon, discussing evaluation issues and describing briefly the various types of summarization techniques. It then examines the characteristics of the medical domain through the different types of medical documents. Finally, it presents and discusses the summarization techniques used so far in the medical domain, referring to the corresponding systems and their characteristics. DISCUSSION AND CONCLUSIONS: The paper discusses thoroughly the promising paths for future research in medical documents summarization. It mainly focuses on the issue of scaling to large collections of documents in various languages and from different media, on personalization issues, on portability to new sub-domains, and on the integration of summarization technology in practical applications. PMID- 15811784 TI - Customization in a unified framework for summarizing medical literature. AB - OBJECTIVE: We present the summarization system in the PErsonalized Retrieval and Summarization of Images, Video and Language (PERSIVAL) medical digital library. Although we discuss the context of our summarization research within the PERSIVAL platform, the primary focus of this article is on strategies to define and generate customized summaries. METHODS AND MATERIAL: Our summarizer employs a unified user model to create a tailored summary of relevant documents for either a physician or lay person. The approach takes advantage of regularities in medical literature text structure and content to fulfill identified user needs. RESULTS: The resulting summaries combine both machine-generated text and extracted text that comes from multiple input documents. Customization includes both group-based modeling for two classes of users, physician and lay person, and individually driven models based on a patient record. CONCLUSIONS: Our research shows that customization is feasible in a medical digital library. PMID- 15811785 TI - A reliable probabilistic sleep stager based on a single EEG signal. AB - OBJECTIVE: We developed a probabilistic continuous sleep stager based on Hidden Markov models using only a single EEG signal. It offers the advantage of being objective by not relying on human scorers, having much finer temporal resolution (1s instead of 30s), and being based on solid probabilistic principles rather than a predefined set of rules (Rechtschaffen & Kales) METHODS AND MATERIAL: Sixty-eight whole night sleep recordings from two different sleep labs are analysed using Gaussian observation Hidden Markov models. RESULTS: Our unsupervised approach detects the cornerstones of human sleep (wakefulness, deep and REM sleep) with around 80% accuracy based on data from a single EEG channel. There are some difficulties in generalizing results across sleep labs. CONCLUSION: Using data from a single electrode is sufficient for reliable continuous sleep staging. Sleep recordings from different sleep labs are not directly comparable. Training of separate models for the sleep labs is necessary. PMID- 15811786 TI - A method for detection of Alzheimer's disease using ICA-enhanced EEG measurements. AB - OBJECTIVE: Many researchers have studied automatic EEG classification and recently a lot of work has been done on artefact-removal from EEG data using independent component analyses (ICA). However, demonstrating that a ICA-processed multichannel EEG measurement becomes more interpretable compared to the raw data (as is usually done in work on ICA-processing of EEG data) does not yet prove that detection of (incipient) anomalies is also better possible after ICA processing. The objective of this study is to show that ICA-preprocessing is useful when constructing a detection system for Alzheimer's disease. METHODS AND MATERIAL: The paper describes a method for detection of EEG patterns indicative of Alzheimer's disease using automatic pattern recognition techniques. Our method incorporates an artefact removal stage based on ICA prior to automatic classification. The method is evaluated on measurements of a length of 8s from two groups of patients, where one group is in an initial stage of the disease (28 patients), whereas the other group is in a more progressed stage (15 patients). Both setups include a control group that should be classified as normal (10 and 21, respectively). RESULTS: Our final classification results for the group with severe Alzheimer's disease are comparable to the best results from literature. We show that ICA-based reduction of artefacts improves classification results for patients in an initial stage. CONCLUSION: We conclude that a more robust detection of Alzheimer's disease related EEG patterns may be obtained by employing ICA as ICA based pre-processing of EEG data can improve classification results for patients in an initial stage of Alzheimer's disease. PMID- 15811787 TI - Applying instance-based techniques to prediction of final outcome in acute stroke. AB - OBJECTIVE: Acute cerebral stroke is a frequent cause of death and the major cause of adult neurological disability in the western world. Thrombolysis is the only established treatment of ischemic stroke; however, its use carries a substantial risk of symptomatic intracerebral hemorrhage. A clinical tool to guide the use of thrombolysis would be very valuable. One of the major goals of such a tool would be the identification of potentially salvageable tissue. This requires an accurate prediction of the extent of infarction if untreated. In this study, we investigate the applicability of highly flexible instance-based (IB) methods for such predictions. METHODS AND MATERIALS: Based on information obtained from magnetic resonance imaging of 14 patients with acute stroke, we explored three different implementations of the IB method: k-NN, Gaussian weighted, and constant radius search classification. Receiver operating characteristics analysis, in particular area under the curve (AUC), was used as performance measure. RESULTS: We found no significant difference (P = 0.48) in performance for the optimal k-NN (k = 164, AUC = 0.814 +/- 0.001) and Gaussian weight (sigma = 0.17, AUC = 0.813 +/- 0.001) implementations, while they were both significantly better (P < 1 x 10(-6) for both) than the constant radius implementation (R = 0.28, AUC = 0.809 +/- 0.001). Qualitative analyses of the distribution of instances in the feature space indicated that non-infarcted instances tends to cluster together while infarcted instances are more dispersed, and that there may not exist a stringent boundary separating infarcted from non-infarcted instances. CONCLUSIONS: This study shows that IB methods can be used, and may be advantageous, for predicting final infarct in patients with acute stroke, but further work must be done to make them clinically applicable. PMID- 15811788 TI - An arrhythmia classification system based on the RR-interval signal. AB - OBJECTIVE: This paper proposes a knowledge-based method for arrhythmic beat classification and arrhythmic episode detection and classification using only the RR-interval signal extracted from ECG recordings. METHODOLOGY: A three RR interval sliding window is used in arrhythmic beat classification algorithm. Classification is performed for four categories of beats: normal, premature ventricular contractions, ventricular flutter/fibrillation and 2 degrees heart block. The beat classification is used as input of a knowledge-based deterministic automaton to achieve arrhythmic episode detection and classification. Six rhythm types are classified: ventricular bigeminy, ventricular trigeminy, ventricular couplet, ventricular tachycardia, ventricular flutter/fibrillation and 2 degrees heart block. RESULTS: The method is evaluated by using the MIT-BIH arrhythmia database. The achieved scores indicate high performance: 98% accuracy for arrhythmic beat classification and 94% accuracy for arrhythmic episode detection and classification. CONCLUSION: The proposed method is advantageous because it uses only the RR-interval signal for arrhythmia beat and episode classification and the results compare well with more complex methods. PMID- 15811789 TI - A classifier based on the artificial neural network approach for cardiologic auscultation in pediatrics. AB - OBJECTIVE: This research work was aimed at developing a reliable screening device for diagnosis of heart murmurs in pediatrics. This is a significant problem in pediatric cardiology because of the high rate of incidence of heart murmurs in this population (reportedly 77-95%), of which only a small fraction arises from congenital heart disease. The screening devices currently available (e.g. chest X ray, electrocardiogram, etc.) suffer from poor sensitivity and specificity in detecting congenital heart disease. Thus, patients with heart murmurs today are frequently assessed by consultation as well with advanced imaging techniques. The most prominent among these is echocardiography. However, echocardiography is expensive and is usually only available in healthcare centers in major cities. Thus, for patients being evaluated with a heart murmur, developing a more accurate screening device is vital to efforts in reducing health care costs. METHODS AND MATERIAL: The data set was collected from incoming pediatrics at the cardiology clinic of The Children's Hospital (Denver, Colorado), on whom echocardiography had been performed to identify congenital heart disease. Recordings of approximately 10-15s duration were made at 44,100Hz and the average record length was approximately 60,000 points. The best three cycles with respect to signal quality sounds were extracted from the original recording. The resulting data comprised 241 examples, of which 88 were examples of innocent murmurs and 153 were examples of pathological murmurs. The selected phonocardiograms were subject to the digital signal processing (DSP) technique of fast Fourier transform (FFT) to extract the energy spectrum in frequency domain. The spectral range was 0-300Hz at a resolution of 1Hz. The processed signals were used to develop statistical classifiers and a classifier based on our in-house artificial neural network (ANN) software. For the latter, we also tried enhancements to the basic ANN scheme. These included a method for setting the decision-threshold and a scheme for consensus-based decision by a committee of experts. RESULTS: Of the different classifiers tested, the ANN-based classifier performed the best. With this classifier, we were able to achieve classification accuracy of 83% sensitivity and 90% specificity in discriminating between innocent and pathological heart murmurs. For the problem of discrimination between innocent murmurs and murmurs of the ventricular septal defect (VSD), the accuracy was higher, with sensitivity of 90% and specificity of 93%. CONCLUSIONS: An ANN-based approach for detection and identification of congenital heart disease in pediatrics from heart murmurs can result in an accurate screening device. Considering that only a simple feature set was used for classification, the results are very encouraging and point out the need for further development using improved feature set with more potent diagnostic variables. PMID- 15811790 TI - Applying spatial distribution analysis techniques to classification of 3D medical images. AB - OBJECTIVE: The objective of this paper is to classify 3D medical images by analyzing spatial distributions to model and characterize the arrangement of the regions of interest (ROIs) in 3D space. METHODS AND MATERIAL: Two methods are proposed for facilitating such classification. The first method uses measures of similarity, such as the Mahalanobis distance and the Kullback-Leibler (KL) divergence, to compute the difference between spatial probability distributions of ROIs in an image of a new subject and each of the considered classes represented by historical data (e.g., normal versus disease class). A new subject is predicted to belong to the class corresponding to the most similar dataset. The second method employs the maximum likelihood (ML) principle to predict the class that most likely produced the dataset of the new subject. RESULTS: The proposed methods have been experimentally evaluated on three datasets: synthetic data (mixtures of Gaussian distributions), realistic lesion-deficit data (generated by a simulator conforming to a clinical study), and functional MRI activation data obtained from a study designed to explore neuroanatomical correlates of semantic processing in Alzheimer's disease (AD). CONCLUSION: Performed experiments demonstrated that the approaches based on the KL divergence and the ML method provide superior accuracy compared to the Mahalanobis distance. The later technique could still be a method of choice when the distributions differ significantly, since it is faster and less complex. The obtained classification accuracy with errors smaller than 1% supports that useful diagnosis assistance could be achieved assuming sufficiently informative historic data and sufficient information on the new subject. PMID- 15811791 TI - Bioinorganic chemistry Coordinate or be coordinated: dynamic relationships between metal ions and biological macromolecules. PMID- 15811792 TI - Probing metal-protein interactions using a de novo design approach. AB - De novo design of metalloproteins provides a valuable tool for understanding the structural constraints and functional attributes of natural biological systems using first principles. This review focuses on recent research aimed primarily at probing the subtle interactions between metals and proteins in designed systems. Considerable attention has focussed on redefining novel design methods used in mimicking natural hemeproteins, mononuclear and dinuclear metallopeptides and functional biological electron-transfer proteins. The present results indicate that the field of metalloprotein design is contributing significantly to the understanding of metals in biology. PMID- 15811793 TI - Metal ions and RNA folding: a highly charged topic with a dynamic future. AB - Metal ions are required to stabilize RNA tertiary structure and to begin the folding process. How different metal ions enable RNAs to fold depends on the electrostatic potential of the RNA and correlated fluctuations in the positions of the ions themselves. Theoretical models, fluorescence spectroscopy, small angle scattering and structural biology reveal that metal ions alter the RNA dynamics and folding transition states. Specifically coordinated divalent metal ions mediate conformational rearrangements within ribozyme active sites. PMID- 15811794 TI - Recent developments in dynamic electrochemical studies of adsorbed enzymes and their active sites. AB - This review outlines recent developments in electrochemical investigations of proteins adsorbed on electrodes. The important point about 'protein film voltammetry' is that it addresses the rates of reactions that occur in enzymes - catalysis, inhibition, electron flow - as a function of potential; in other words, it introduces the 'potential dimension' into enzyme kinetics. Some surprisingly subtle, yet significant observations are made, including demonstration of a special role for Mo(V) in the catalytic cycle of Mo enzymes, quantitation of the catalytic bias in multi-centred enzymes such as mitochondrial Complex I, insight into mechanisms of proton transfer in enzymes, and properties of proteins that are covalently attached directly to a gold surface. PMID- 15811795 TI - Design and engineering of metalloproteins containing unnatural amino acids or non native metal-containing cofactors. AB - An emerging branch of metalloprotein design and engineering is on the horizon, where unnatural amino acids or non-native metal-containing cofactors are employed in the design and engineering process. These endeavors have been shown to be quite effective in elucidating the precise roles of key residues in protein structures and functions, in providing guiding principles on protein design, in fine-tuning the protein properties to an unprecedented level, and in expanding the repertoire of protein functionalities, and thus its range of applications. PMID- 15811796 TI - Mapping nucleic acid structure by hydroxyl radical cleavage. AB - Hydroxyl radical footprinting is a widely used method for following the folding of RNA molecules in solution. This method has the unique ability to provide experimental information on the solvent accessibility of each nucleotide in an RNA molecule, so that the folding of all domains of the RNA species can be followed simultaneously at single-nucleotide resolution. In recent work, hydroxyl radical footprinting has been used, often in combination with other global measures of structure, to work out detailed folding pathways and three dimensional structures for increasingly large and complicated RNA molecules. These include synthetic ribozymes, and group I and group II ribozymes, from yeast, the Azoarcus cyanobacterium and Tetrahymena thermophila. Advances have been made in methods for analysis of hydroxyl radical data, so that the large datasets that result from kinetic folding experiments can be analyzed in a semi automated and quantitative manner. PMID- 15811797 TI - Metal complex-DNA interactions: from transcription inhibition to photoactivated cleavage. AB - Metal ions and complexes, because of their cationic character, three-dimensional structural profiles, and propensity for performing hydrolysis, redox, or photoreactions, have a natural aptitude for interacting with DNA. Indeed, the need for cellular regulation of DNA led to the evolution of metallonucleases to catalyze and repair DNA strand breaks. Moreover, inorganic constructs such as cisplatin and bimetallic rhodium acetate exert antitumor activity by inner-sphere coordination to DNA. Because binding and cleavage of DNA is at the heart of cellular transcription and translation, it is an obvious target for therapeutic intervention and the development of diagnostic structural probes. To this end, new metal complexes have been designed that utilize or create open coordination positions for DNA binding and hydrolysis, generate reactive oxygen-containing species or other radicals for DNA oxidation, or perform direct redox reactions with DNA. The recent emerging themes are the development of bifunctional architectures containing multiple metal-binding or reactive sites, specialized ligand implementation, or incorporation of site-specific targeting substructures. This review describes their employment in novel reaction strategies that do not require bimolecular cofactors and as site-specific probes or cleavage agents. PMID- 15811798 TI - A role for iron-sulfur clusters in DNA repair. AB - The presence of 4Fe-4S clusters in enzymes involved in DNA repair has posed the question of the role of these intricate cofactors in damaged DNA recognition and repair. It is particularly intriguing that base excision repair glycosylases that remove a wide variety of damaged bases, and also have vastly different sequences and structures, have been found to contain this cofactor. The accumulating biochemical and structural evidence indicates that the region supported by the cluster is intimately involved in DNA binding, and that such binding interactions impact catalysis of base removal. Recent evidence has also established that binding of the glycosylases to DNA facilitates oxidation of the [4Fe-4S](2+) cluster to the [4Fe-4S](3+) form. Notably, the measured redox potentials for a variety of 4Fe-4S cluster-containing glycosylases are remarkably similar. Based on this DNA-mediated redox behavior, it has been suggested that this property may be used to enhance the activity of these enzymes by facilitating damaged DNA location. PMID- 15811799 TI - Dioxygen activation by copper, heme and non-heme iron enzymes: comparison of electronic structures and reactivities. AB - Enzymes containing heme, non-heme iron and copper active sites play important roles in the activation of dioxygen for substrate oxidation. One key reaction step is CH bond cleavage through H-atom abstraction. On the basis of the ligand environment and the redox properties of the metal, these enzymes employ different methods of dioxygen activation. Heme enzymes are able to stabilize the very reactive iron(IV)-oxo porphyrin-radical intermediate. This is generally not accessible for non-heme iron systems, which can instead use low-spin ferric hydroperoxo and iron(IV)-oxo species as reactive oxidants. Copper enzymes employ still a different strategy and achieve H-atom abstraction potentially through a superoxo intermediate. This review compares and contrasts the electronic structures and reactivities of these various oxygen intermediates. PMID- 15811800 TI - Biocatalysis and biotransformation New technologies, enzymes and challenges. PMID- 15811801 TI - Recent progress towards the application of hyperthermophiles and their enzymes. AB - The discovery of extremophiles has drastically changed our understanding towards the diversity of life itself and the conditions under which it can be sustained. Extremophiles have evolved to withstand and multiply under the extremes of temperature, pressure, pH and salinity. Hyperthermophiles are the group that have adapted to high temperature; many have been found to grow at temperatures above the boiling point of water. This review focuses on recent advances in application based research on hyperthermophiles and their thermostable enzymes. PMID- 15811802 TI - Whole organism biocatalysis. AB - The use of biocatalysts for the industrial synthesis of chemicals has been attracting much attention as an environment-friendly synthetic method. Microbial cells play a leading role in 'chemo-enzymatic synthesis' because of their great diversity. Several microbes with unique catalytic abilities have been found through intensive screening and put to practical use. Besides, advanced molecular biological techniques are powerful tools for developing more satisfactory biocatalysts. PMID- 15811803 TI - New enzymes for biotransformations. AB - Several novel bioprocesses that have little or no counterpart in traditional methodology have recently been reported. The stereoselective and enantioselective hydrolysis of sec-alkyl sulfate esters by alkyl sulfatases proceeds with inversion of configuration and furnishes a homochiral product mixture. Haloalcohol dehalogenases were shown to accept various non-natural nucleophiles, such as azide, cyanide and nitrite for the asymmetric opening of epoxides giving rise to the corresponding azido-, cyano-, and nitro-alcohols as non-natural products. Asymmetric carbon-carbon bond formation via the acyloin- and benzoin reaction was successfully catalyzed in water by novel lyases, such as benzoylformate decarboxylase and benzaldehyde lyase. New methods for the production of chiral nonracemic alpha-L-amino acids and amines were recently reported. Enantioselective stereoinversion of racemic alpha-aryl- and alpha aryloxycarboxylic acids via epimerase-catalyzed inversion led to a single stereoisomeric product from the racemate. PMID- 15811804 TI - Advances in biocatalytic synthesis of pharmaceutical intermediates. AB - Biocatalytic synthesis of enantiomerically pure compounds for pharmaceutical intermediates is gaining momentum. This is the result of advances in genomics, screening and evolution technologies leading to the increased availability of new and robust biocatalysts suited for industrial-scale application, and is stimulated by an increased demand for catalysts that are able to address the increased complexity of active pharmaceutical ingredients. The vast majority of biotransformation reactions for the manufacturing of optically active pharmaceutical intermediates are still based on enantioselective ketone reductions and enantiospecific hydrolyses. This review aims to point at alternative reaction types and integrated multi-enzymatic steps that are emerging in large-scale applications. PMID- 15811805 TI - Enhancing catalytic promiscuity for biocatalysis. AB - Catalytic promiscuity - the ability of a single active site to catalyse more than one chemical transformation - has a natural role in evolution and occasionally in biosynthesis of secondary metabolites. Catalytic promiscuity is more widespread than often recognized. Recent success in adding and enhancing such catalytic activities by protein engineering suggests new potential applications in enzyme catalyzed organic synthesis. PMID- 15811806 TI - Predicting enzyme function from protein sequence. AB - There are two main reasons to try to predict an enzyme's function from its sequence. The first is to identify the components and thus the functional capabilities of an organism, the second is to create enzymes with specific properties. Genomics, expression analysis, proteomics and metabonomics are largely directed towards understanding how information flows from DNA sequence to protein functions within an organism. This review focuses on information flow in the opposite direction: the applicability of what is being learned from natural enzymes to improve methods for catalyst design. PMID- 15811807 TI - High-throughput screens and selections of enzyme-encoding genes. AB - The availability of vast gene repertoires from both natural sources (genomic and cDNA libraries) and artificial sources (gene libraries) demands the development and application of novel technologies that enable the screening or selection of large libraries for a variety of enzymatic activities. We describe recent developments in the selection of enzyme-coding genes for directed evolution and functional genomics. We focus on HTS approaches that enable selection from large libraries (>10(6) gene variants) with relatively humble means (i.e. non-robotic systems), and on in vitro compartmentalization in particular. PMID- 15811808 TI - Immobilised enzymes: science or art? AB - Enzyme immobilisation is experiencing an important transition. Combinatorial approaches are increasingly applied in the design of robust immobilised enzymes by rational combination of fundamental immobilisation techniques (i.e. non covalent adsorption, covalent binding, entrapment and encapsulation) or with other relevant technologies. The objective is to solve specific problems that cannot be solved by one of these basic immobilisation techniques. PMID- 15811809 TI - Reinventing the wheels. PMID- 15811811 TI - Overlooked in Fallon? PMID- 15811813 TI - "Arsenic in food": opinion parading as science. PMID- 15811815 TI - "Sex and ceruloplasmin modulate the response to copper . . .". PMID- 15811816 TI - Iraq's toxic shipwrecks. PMID- 15811817 TI - MIT toxicogenomics research program. PMID- 15811818 TI - Heavy traffic ahead: car culture accelerates. PMID- 15811819 TI - Driving up the cost of clean air. PMID- 15811820 TI - Here comes hypercar! PMID- 15811822 TI - Pulmonary effects of indoor- and outdoor-generated particles in children with asthma. AB - Most particulate matter (PM) health effects studies use outdoor (ambient) PM as a surrogate for personal exposure. However, people spend most of their time indoors exposed to a combination of indoor-generated particles and ambient particles that have infiltrated. Thus, it is important to investigate the differential health effects of indoor- and ambient-generated particles. We combined our recently adapted recursive model and a predictive model for estimating infiltration efficiency to separate personal exposure (E) to PM2.5 (PM with aerodynamic diameter < or = 2.5 microm) into its indoor-generated (Eig) and ambient-generated (Eag) components for 19 children with asthma. We then compared Eig and Eag to changes in exhaled nitric oxide (eNO), a marker of airway inflammation. Based on the recursive model with a sample size of eight children, Eag was marginally associated with increases in eNO [5.6 ppb per 10-microg/m3 increase in PM2.5; 95% confidence interval (CI), -0.6 to 11.9; p = 0.08]. Eig was not associated with eNO (-0.19 ppb change per 10 microg/m3). Our predictive model allowed us to estimate Eag and Eig for all 19 children. For those combined estimates, only Eag was significantly associated with an increase in eNO (Eag: 5.0 ppb per 10 microg/m3 increase in PM2.5; 95% CI, 0.3 to 9.7; p = 0.04; Eig: 3.3 ppb per 10 microg/m3 increase in PM2.5; 95% CI, -1.1 to 7.7; p = 0.15). Effects were seen only in children who were not using corticosteroid therapy. We conclude that the ambient-generated component of PM2.5 exposure is consistently associated with increases in eNO and the indoor-generated component is less strongly associated with eNO. PMID- 15811823 TI - Alkyl phenols and diethylhexyl phthalate in tissues of sheep grazing pastures fertilized with sewage sludge or inorganic fertilizer. AB - We studied selected tissues from ewes and their lambs that were grazing pastures fertilized with either sewage sludge (treated) or inorganic fertilizer (control) and determined concentrations of alkylphenols and phthalates in these tissues. Mean tissue concentrations of alkylphenols were relatively low (< 10-400 microg/kg) in all animals and tissues. Phthalates were detected in tissues of both control and treated animals at relatively high concentrations (> 20,000 microg/kg in many tissue samples). The use of sludge as a fertilizer was not associated with consistently increased concentrations of either alkylphenols or phthalates in the tissues of animals grazing treated pastures relative to levels in control animal tissues. Concentrations of the two classes of chemicals differed but were of a similar order of magnitude in liver and muscle as well as in fat. Concentrations of each class of compound were broadly similar in tissues derived from ewes and lambs. Although there were significant differences (p < 0.01 or p < 0.001) between years (cohorts) in mean tissue concentrations of both nonylphenol (NP) and phthalate in each of the tissues from both ewes and lambs, the differences were not attributable to either the age (6 months or 5 years) of the animal or the duration of exposure to treatments. Octylphenol concentrations were generally undetectable. There was no consistent cumulative outcome of prolonged exposure on the tissue concentrations of either class of pollutant in any ewe tissue. Mean tissue concentrations of phthalate were higher (p < 0.001) in the liver and kidney fat of male compared with female lambs. We suggest that the addition of sewage sludge to pasture is unlikely to cause large increases in tissue concentrations of NP and phthalates in sheep and other animals with broadly similar diets and digestive systems (i.e., domestic ruminants) grazing such pasture. PMID- 15811824 TI - Concern about environmental pollution: how much difference do race and ethnicity make? A New Jersey case study. AB - A survey conducted among 1,513 residents of New Jersey during March-May 2004 showed that non-Hispanic black, non-Hispanic white, and English-speaking Hispanic Americans were significantly more concerned about environmental pollution problems than were Asian Americans and Spanish-language Hispanic Americans. For example, an average of > 40% of the first three groups was very concerned about New Jersey's environmental problems, compared with 15% of the last two populations. There were also racial/ethnic differences among these groups in their desire for government action to protect the environment and in their personal support of the environmental movement. Regression analyses suggest that the 1970s and 1980s model of core support for environmental protection from white, female, young, educated, and politically liberal people has largely, but not completely, continued among non-Hispanic white, non-Hispanic black, and English-language Hispanic populations. But these demographic pointers do not hold for Asian and Spanish-language Hispanic Americans, except indicating more support among the more formally educated. The last two groups are the two fastest-growing subpopulations in the United States, and although acculturation may slowly increase their concern about environmental pollution, it is more prudent for proponents of environmental protection not to wait and instead to try to better understand the environmental perceptions of these groups. PMID- 15811825 TI - Ambient air pollution and pregnancy outcomes: a review of the literature. AB - Over the last decade or so, a large number of studies have investigated the possible adverse effects of ambient air pollution on birth outcomes. We reviewed these studies, which were identified by a systematic search of the main scientific databases. Virtually all reviewed studies were population based, with information on exposure to air pollution derived from routine monitoring sources. Overall, there is evidence implicating air pollution in adverse effects on different birth outcomes, but the strength of the evidence differs between outcomes. The evidence is sufficient to infer a causal relationship between particulate air pollution and respiratory deaths in the postneonatal period. For air pollution and birth weight the evidence suggests causality, but further studies are needed to confirm an effect and its size and to clarify the most vulnerable period of pregnancy and the role of different pollutants. For preterm births and intrauterine growth retardation (IUGR) the evidence as yet is insufficient to infer causality, but the available evidence justifies further studies. Molecular epidemiologic studies suggest possible biologic mechanisms for the effect on birth weight, premature birth, and IUGR and support the view that the relation between pollution and these birth outcomes is genuine. For birth defects, the evidence base so far is insufficient to draw conclusions. In terms of exposure to specific pollutants, particulates seem the most important for infant deaths, and the effect on IUGR seems linked to polycyclic aromatic hydrocarbons, but the existing evidence does not allow precise identification of the different pollutants or the timing of exposure that can result in adverse pregnancy outcomes. PMID- 15811826 TI - Chemical mixtures: considering the evolution of toxicology and chemical assessment. AB - The assessment of chemical mixtures is a complex topic for toxicologists, regulators, and the public. In this article the linkage between the science of toxicology and the needs of governmental regulatory agencies in the United States is explored through an overview of environmental regulations enacted over the past century and a brief history of modern toxicology. One of the goals of this overview is to encourage both regulators and scientists to consider the benefits and limitations of this science-regulatory relationship as they tackle existing issues such as chemical mixtures. It is clear that a) over the past 100 years chemical regulation and toxicologic research, have in large part, shared a common emphasis on characterization and regulation of individual chemicals. But chemical mixtures have been, and continue to be, evaluated at hazardous waste sites around the United States. For this reason the current U.S. Environmental Protection Agency guidelines for chemical mixtures assessment are also reviewed. These guidelines highlight the current practice of mixtures assessment, which relies primarily on the existing single-chemical database. It is also clear that b) the science and assessment of chemical mixtures are moving forward through the combined efforts of regulatory agencies and scientists from a broad range of disciplines, including toxicology. Because toxicology is at this exciting crossroads, particular attention should be paid to the forces (e.g., public demands, regulatory needs, funding, academic interests) that both promote and limit the growth of this expanding discipline. PMID- 15811827 TI - Urinary concentrations of bisphenol A and 4-nonylphenol in a human reference population. AB - Bisphenol A (BPA) is used to manufacture polycarbonate plastic and epoxy resins, which are used in baby bottles, as protective coatings on food containers, and for composites and sealants in dentistry. 4-Nonylphenol (NP) is used to make nonylphenol ethoxylates, nonionic surfactants applied as emulsifying, wetting, dispersing, or stabilizing agents in industrial, agricultural, and domestic consumer products. The potential for human exposure to BPA and NP is high because of their widespread use. We measured BPA and NP in archived urine samples from a reference population of 394 adults in the United States using isotope-dilution gas chromatography/mass spectrometry. The concentration ranges of BPA and NP were similar to those observed in other human populations. BPA was detected in 95% of the samples examined at concentrations > or = 0.1 microg/L urine; the geometric mean and median concentrations were 1.33 microg/L (1.36 microg/g creatinine) and 1.28 microg/L (1.32 microg/g creatinine), respectively; the 95th percentile concentration was 5.18 microg/L (7.95 microg/g creatinine). NP was detected in 51% of the samples examined > or = 0.1 microg/L. The median and 95th percentile concentrations were < 0.1 microg/L and 1.57 microg/L (1.39 microg/g creatinine), respectively. The frequent detection of BPA suggests widespread exposure to this compound in residents of the United States. The lower frequency of detection of NP than of BPA could be explained by a lower exposure of humans to NP, by different pharmacokinetic factors (i.e., absorption, distribution, metabolism, elimination), by the fact that 4-n-nonylphenol--the measured NP isomer- represents a small percentage of the NP used in commercial mixtures, or a combination of all of the above. Additional research is needed to determine the best urinary biomarker(s) to assess exposure to NP. Despite the sample population's nonrepresentativeness of the U.S. population (although sample weights were used to improve the extent to which the results represent the U.S. population) and relatively small size, this study provides the first reference range of human internal dose levels of BPA and NP in a demographically diverse human population. PMID- 15811828 TI - Effects of ammonium perchlorate on thyroid function in developing fathead minnows, Pimephales promelas. AB - Perchlorate is a known environmental contaminant, largely due to widespread military use as a propellant. Perchlorate acts pharmacologically as a competitive inhibitor of thyroidal iodide uptake in mammals, but the impacts of perchlorate contamination in aquatic ecosystems and, in particular, the effects on fish are unclear. Our studies aimed to investigate the effects of concentrations of ammonium perchlorate that can occur in the environment (1, 10, and 100 mg/L) on the development of fathead minnows, Pimephales promelas. For these studies, exposures started with embryos of < 24-hr postfertilization and were terminated after 28 days. Serial sectioning of thyroid follicles showed thyroid hyperplasia with increased follicular epithelial cell height and reduced colloid in all groups of fish that had been exposed to perchlorate for 28 days, compared with control fish. Whole-body thyroxine (T4) content (a measure of total circulating T4 in fish exposed to 100 mg/L perchlorate was elevated compared with the T4 content of control fish, but 3,5,3-triiodothyronine (T3) content was not significantly affected in any exposure group. Despite the apparent regulation of T3, after 28 days of exposure to ammonium perchlorate, fish exposed to the two higher levels (10 and 100 mg/L) were developmentally retarded, with a lack of scales and poor pigmentation, and significantly lower wet weight and standard length than were control fish. Our study indicates that environmental levels of ammonium perchlorate affect thyroid function in fish and that in the early life stages these effects may be associated with developmental retardation. PMID- 15811829 TI - The precautionary principle and risk perception: experimental studies in the EMF area. AB - Possible adverse health effects due to electromagnetic fields (EMFs) from cellular phones and base stations present a major public health issue across Europe. Because scientists cannot exclude that EMFs may cause health problems, the application of the precautionary principle is debated heavily. By considering precautionary measures, political decision makers hope to cope with public fears about EMFs. We present results from two experimental studies that indicate that precautionary measures may trigger concerns, amplify EMF-related risk perceptions, and lower trust in public health protection. Such impacts, questioning common expectations, should be considered in decisions about precautionary measures. PMID- 15811830 TI - The World Trade Center residents' respiratory health study: new-onset respiratory symptoms and pulmonary function. AB - The destruction of the World Trade Center (WTC) on 11 September 2001 in New York City resulted in the massive release of pulverized dust and combustion products. The dust and smoke settled in the surrounding area, which encompassed a large residential community. We hypothesized that previously normal residents in the community surrounding the former WTC would have an increased incidence of persistent respiratory symptoms and abnormalities in screening spirometry. A hybrid cross-sectional and retrospective cohort study using a symptom-based questionnaire and onsite screening spirometry in residents in an exposed area and in a control area was performed 12 +/- 4 months after the collapse. Surveys were analyzed from 2,812 residents. New-onset respiratory symptoms were described by 55.8% of residents in the exposed area, compared with 20.1% in the control area after the event. Persistent new-onset symptoms were identified in 26.4 versus 7.5% of residents in the exposed area versus control area, respectively. No differences in screening spirometry between the groups were detected. A small pilot study suggested the possibility of an increase in bronchial hyperresponsiveness in exposed participants with persistent symptoms. The data demonstrate an increased rate of new-onset and persistent respiratory health effects in residents near the former WTC compared with a control population. PMID- 15811831 TI - Effect of environmental tobacco smoke on levels of urinary hormone markers. AB - Our recent study showed a dose-response relationship between environmental tobacco smoke (ETS) and the risk of early pregnancy loss. Smoking is known to affect female reproductive hormones. We explored whether ETS affects reproductive hormone profiles as characterized by urinary pregnanediol-3-glucuronide (PdG) and estrone conjugate (E1C) levels. We prospectively studied 371 healthy newly married nonsmoking women in China who intended to conceive and had stopped contraception. Daily records of vaginal bleeding, active and passive cigarette smoking, and daily first-morning urine specimens were collected for up to 1 year or until a clinical pregnancy was achieved. We determined the day of ovulation for each menstrual cycle. The effects of ETS exposure on daily urinary PdG and E1C levels in a +/-10 day window around the day of ovulation were analyzed for conception and nonconception cycles, respectively. Our analysis included 344 nonconception cycles and 329 conception cycles. In nonconception cycles, cycles with ETS exposure had significantly lower urinary E1C levels (beta = -0.43, SE = 0.08, p < 0.001 in log scale) compared with the cycles without ETS exposure. There was no significant difference in urinary PdG levels in cycles having ETS exposure (beta = -0.07, SE = 0.15, p = 0.637 in log scale) compared with no ETS exposure. Among conception cycles, there were no significant differences in E1C and PdG levels between ETS exposure and nonexposure. In conclusion, ETS exposure was associated with significantly lower urinary E1C levels among nonconception cycles, suggesting that the adverse reproductive effect of ETS may act partly through its antiestrogen effects. PMID- 15811832 TI - Chlorination disinfection by-products and pancreatic cancer risk. AB - Chlorination disinfection by-products (CDBPs) are produced during the treatment of water with chlorine to remove bacterial contamination. CDBPs have been associated with an increased risk of bladder cancer. There is also some evidence that they may increase the risk of pancreatic cancer. We report results from a population-based case-control study of 486 incident cases of pancreatic cancer and 3,596 age- and sex-matched controls. Exposure to chlorination by-products was estimated by linking lifetime residential histories to two different databases containing information on CDBP levels in municipal water supplies. Logistic regression analysis found no evidence of increased pancreatic cancer risk at higher CDBP concentrations (all odds ratios < 1.3). Null findings were also obtained assuming a latency period for pancreatic cancer induction of 3, 8, or 13 years. PMID- 15811833 TI - Evidence of interaction between polychlorinated biphenyls and phthalates in relation to human sperm motility. AB - Previously, we reported evidence of inverse associations between exposure to some polychlorinated biphenyls (PCBs) and some phthalate monoesters in relation to semen parameters, specifically sperm motility. Because humans are exposed to both phthalates and PCBs and because experimental studies suggest that PCBs may interact with glucuronidative enzymes that are responsible for phthalate metabolism, we explored the potential interaction between phthalates and PCBs in relation to human semen quality. We studied 303 men who were partners in subfertile couples seeking infertility diagnosis from the andrology laboratory at Massachusetts General Hospital. Semen parameters were dichotomized based on World Health Organization reference values, and phthalate and PCB levels were dichotomized at their respective medians. After adjusting for age and abstinence time, for below reference sperm motility there was a greater than additive interaction between monobenzyl phthalate and PCB-153 [relative excess risk due to interaction (RERI) = 1.40; 95% confidence interval (CI), 0.41-3.22], sum of PCBs (RERI = 1.24; 95% CI, 0.15-2.94), and cytochrome P450 (CYP450)-inducing PCBs (RERI = 1.30; 95% CI, 0.21-3.06). For below-reference sperm motility, there was also a greater than additive interaction between monobutyl phthalate (MBP) and PCB-153 (RERI = 1.42; 95% CI, 0.09-3.76) and CYP450-inducing PCBs (RERI = 1.87; 95% CI, 0.56-4.52) and a suggestive interaction between MBP and sum of PCBs (RERI = 1.35; 95% CI, -0.11 to 3.48). In conclusion, because there are important risk assessment and public health implications of interactions between these two ubiquitous classes of compounds, further studies need to be conducted to confirm these results and identify potential mechanisms of interactions. PMID- 15811834 TI - Xenoestrogens at picomolar to nanomolar concentrations trigger membrane estrogen receptor-alpha-mediated Ca2+ fluxes and prolactin release in GH3/B6 pituitary tumor cells. AB - Xenoestrogens (XEs) are widespread in our environment and are known to have deleterious effects in animal (and perhaps human) populations. Acting as inappropriate estrogens, XEs are thought to interfere with endogenous estrogens such as estradiol (E2) to disrupt normal estrogenic signaling. We investigated the effects of E2 versus several XEs representing organochlorine pesticides (dieldrin, endosulfan, o',p'-dichlorodiphenylethylene), plastics manufacturing by products/detergents (nonylphenol, bisphenol A), a phytoestrogen (coumestrol), and a synthetic estrogen (diethylstilbestrol) on the pituitary tumor cell subline GH3/B6/F10, previously selected for expression of high levels of membrane estrogen receptor-alpha. Picomolar to nanomolar concentrations of both E2 and XEs caused intracellular Ca2+ changes within 30 sec of administration. Each XE produced a unique temporal pattern of Ca2+ elevation. Removing Ca2+ from the extracellular solution abolished both spontaneous and XE-induced intracellular Ca2+ changes, as did 10 microM nifedipine. This suggests that XEs mediate their actions via voltage-dependent L-type Ca2+ channels in the plasma membrane. None of the Ca2+ fluxes came from intracellular Ca2+ stores. E2 and each XE also caused unique time- and concentration-dependent patterns of prolactin (PRL) secretion that were largely complete within 3 min of administration. PRL secretion was also blocked by nifedipine, demonstrating a correlation between Ca2+ influx and PRL secretion. These data indicate that at very low concentrations, XEs mediate membrane-initiated intracellular CCa2+ increases resulting in PRL secretion via a mechanism similar to that for E2, but with distinct patterns and potencies that could explain their abilities to disrupt endocrine functions. PMID- 15811835 TI - Repolarization changes induced by air pollution in ischemic heart disease patients. AB - Epidemiologic studies report associations between particulate air pollution and cardiovascular morbidity and mortality, but the underlying pathophysiologic mechanisms are still unclear. We tested the hypothesis that patients with preexisting coronary heart disease experience changes in the repolarization parameters in association with rising concentrations of air pollution. A prospective panel study was conducted in Erfurt, East Germany, with 12 repeated electrocardiogram (ECG) recordings in 56 males with ischemic heart disease. Hourly particulate and gaseous air pollution and meteorologic data were acquired. The following ECG parameters reflecting myocardial substrate and vulnerability were measured: QT duration, T-wave amplitude, T-wave complexity, and variability of T-wave complexity. Fixed effect regression analysis was used adjusting for subject, trend, weekday, and meteorology. The analysis showed a significant increase in QT duration in response to exposure to organic carbon; a significant decrease in T-wave amplitude with exposure to ultrafine, accumulation mode, and PM2.5 particles (particles < 2.5 microm in aerodynamic diameter); and a corresponding significant increase of T-wave complexity in association with PM2.5 particles for the 24 hr before ECG recordings. Variability of T-wave complexity showed a significant increase with organic and elemental carbon in the same time interval. This study provides evidence suggesting an immediate effect of air pollution on repolarization duration, morphology, and variability representing myocardial substrate and vulnerability, key factors in the mechanisms of cardiac death. PMID- 15811836 TI - The induction of vascular endothelial growth factor by ultrafine carbon black contributes to the increase of alveolar-capillary permeability. AB - Ultrafine carbon black (ufCB) can cause proinflammatory response and increase alveolar-capillary permeability. However, the mechanism underlying the increased permeability is not well characterized. Vascular endothelial growth factor (VEGF) is originally recognized as a vascular permeability factor. Oxidative stress generated by hydrogen peroxide (H2O2) stimulates VEGF gene expression. The purpose of this study was to explore the role of VEGF in ufCB-induced alveolar capillary permeability. Intratracheal instillation of 200 microg ufCB in mice caused a significant and sustained increase of total proteins in bronchoalveolar lavage (BAL) fluid, with the maximal increase at 21 hr postinstillation. The influx of neutrophils did not significantly increase until 16 hr. It reached the highest level at 21 hr and returned to the basal level by 42 hr. Tumor necrosis factor-alpha was significantly elevated only at 4 hr. ufCB induced significant increases of VEGF in BAL fluid throughout the study period, with the peak increase at 16 hr. The nonsecreted isoform VEGF188 was not altered after 16 hr of exposure to ufCB. Moreover, there was a strong correlation between VEGF and total proteins in BAL fluid (R2 = 0.7352, p < 0.01). In vivo study supported the role of reactive oxygen species (ROSs) in ufCB-induced VEGF release and protein leakage. The involvement of ROSs was strengthened by the fact that interventions with N-acetylcysteine prevented ufCB-induced generation of ROSs and VEGF in vitro. Our study for the first time demonstrates that ufCB induces the production of VEGF, which is associated with the increase of alveolar-capillary permeability. The induction of VEGF by ufCB acts through an ROS-dependent pathway. PMID- 15811837 TI - Domoic acid transfer to milk: evaluation of a potential route of neonatal exposure. AB - Domoic acid (DA), produced by the diatom genus Pseudo-nitzschia, is a glutamate analog and a neurotoxin in humans. During diatom blooms, DA can contaminate filter-feeding organisms, such as shellfish, and can be transferred by ingestion to higher trophic levels. Several intoxication events involving both humans and various marine mammals have been attributed to DA. Affected organisms show neurological symptoms such as seizures, ataxia, headweaving, and stereotypic scratching, as well as prolonged deficits in memory and learning. Neonatal animals have been shown to be substantially more sensitive to DA than adults. However, it has not been demonstrated whether DA can be transferred to nursing young from DA-exposed mothers. This study demonstrates transfer of DA from spiked milk (0.3 and 1.0 mg/kg) to the plasma of nursing neonatal rats and an overall longer DA retention in milk than in plasma after 8 hr in exposed dams. DA was detectable in milk up to 24 hr after exposure (1.0 mg/kg) of the mothers, although the amount of DA transferred to milk after exposure was not sufficient to cause acute symptoms in neonates. PMID- 15811838 TI - Coupling aggressive mass removal with microbial reductive dechlorination for remediation of DNAPL source zones: a review and assessment. AB - The infiltration of dense non-aqueous-phase liquids (DNAPLs) into the saturated subsurface typically produces a highly contaminated zone that serves as a long term source of dissolved-phase groundwater contamination. Applications of aggressive physical-chemical technologies to such source zones may remove > 90% of the contaminant mass under favorable conditions. The remaining contaminant mass, however, can create a rebounding of aqueous-phase concentrations within the treated zone. Stimulation of microbial reductive dechlorination within the source zone after aggressive mass removal has recently been proposed as a promising staged-treatment remediation technology for transforming the remaining contaminant mass. This article reviews available laboratory and field evidence that supports the development of a treatment strategy that combines aggressive source-zone removal technologies with subsequent promotion of sustained microbial reductive dechlorination. Physical-chemical source-zone treatment technologies compatible with posttreatment stimulation of microbial activity are identified, and studies examining the requirements and controls (i.e., limits) of reductive dechlorination of chlorinated ethenes are investigated. Illustrative calculations are presented to explore the potential effects of source-zone management alternatives. Results suggest that, for the favorable conditions assumed in these calculations (i.e., statistical homogeneity of aquifer properties, known source zone DNAPL distribution, and successful bioenhancement in the source zone), source longevity may be reduced by as much as an order of magnitude when physical chemical source-zone treatment is coupled with reductive dechlorination. PMID- 15811839 TI - Impact of occupational exposure on lead levels in women. AB - In 1994, 207 women participated in a study designed to examine the effects of occupational exposure and various lifestyle factors on bone and blood lead levels. In vivo measurements of Pb concentrations in tibia were performed by X ray fluorescence. All 108 former smelter employees and 99 referents provided blood samples and answered a questionnaire on lifestyle characteristics and the relevant medical history. Lead concentrations in tibia and blood were significantly higher in the exposed group. The difference in mean bone Pb concentrations of the two groups is markedly greater than the difference in the mean blood Pb concentrations, supporting the view that bone Pb measurements are a more reliable determinant of Pb body burden. Chronic exposure did not result in any statistically significant differences in adverse pregnancy outcomes. A significantly lower age at the onset of menopause in occupationally exposed women may suggest that Pb causes adverse changes in the pattern of estrus and menses. The exposed women had lower bone Pb concentrations than those found in most studies on predominantly male workers. Blood Pb concentrations remain increased in women long after the cessation of occupational exposure, reflecting the importance of the endogenous exposure. The endogenous exposure relation found for postmenopausal exposed women is consistent with data on male smelter workers, whereas the relation found for premenopausal women is significantly lower. This suggests that sex plays an important role in the metabolism of lead, and current models of exposure extrapolated from male data may be inappropriate for use on women. PMID- 15811840 TI - Respiratory morbidity in office workers in a water-damaged building. AB - We conducted a study on building-related respiratory disease and associated social impact in an office building with water incursions in the northeastern United States. An initial questionnaire had 67% participation (888/1,327). Compared with the U.S. adult population, prevalence ratios were 2.2-2.5 for wheezing, lifetime asthma, and current asthma, 3.3 for adult-onset asthma, and 3.4 for symptoms improving away from work (p < 0.05). Two-thirds (66/103) of the adult-onset asthma arose after occupancy, with an incidence rate of 1.9/1,000 person-years before building occupancy and 14.5/1,000 person-years after building occupancy. We conducted a second survey on 140 respiratory cases, 63 subjects with fewer symptoms, and 44 comparison subjects. Health-related quality of life decreased with increasing severity of respiratory symptoms and in those with work related symptoms. Symptom status was not associated with job satisfaction or how often jobs required hard work. Respiratory health problems accounted for one third of sick leave, and respiratory cases with work-related symptoms had more respiratory sick days than those without work-related symptoms (9.4 vs. 2.4 days/year; p < 0.01). Abnormal lung function and/or breathing medication use was found in 67% of respiratory cases, in 38% of participants with fewer symptoms, and in 11% of the comparison group (p < 0.01), with similar results in never smokers. Postoccupancy-onset asthma was associated with less atopy than preoccupancy-onset asthma. Occupancy of the water-damaged building was associated with onset and exacerbation of respiratory conditions, confirmed by objective medical tests. The morbidity and lost work time burdened both employees and employers. PMID- 15811841 TI - Intoxication from an accidentally ingested lead shot retained in the gastrointestinal tract. AB - A 45-year-old woman was referred to the Department of Occupational and Environmental Health in January 2002 because of increased blood lead concentrations of unknown origin. She suffered from malaise, fatigue, and diffuse gastrointestinal symptoms. She had a blood lead level of 550 microg/L (normal range < 40 microg/L). The patient had not been occupationally exposed to lead, and no potential lead sources, such as food products or lead-glazed pottery, could be identified. Her food habits were normal, but she did consume game occasionally. Clinical examination, including standard neurologic examination, was normal. No anemia was present. Laboratory tests showed an increased excretion of lead in the urine, but there were no signs of microproteinuria. An abdominal X ray in October 2002 revealed a 6-mm rounded metal object in the colon ascendens. Before the object could be further localized, the patient contracted winter vomiting disease (gastroenteritis) and the metal object was spontaneously released from the colon during a diarrhea attack. The object was a lead shot pellet, possibly but not normally used in Sweden for hunting wild boar or roe deer. Blood lead levels slowly decreased. Nine months later the patient's blood lead levels were almost normal (approximately 70 microg/L) and her symptoms had almost completely disappeared. In this case, a rare source of lead exposure was found. In investigations of blood lead elevations of unknown origin, we recommend abdominal X ray in parallel with repeated blood lead determinations. PMID- 15811842 TI - Assessing exposure to organophosphorus pesticides by biomonitoring in epidemiologic studies of birth outcomes. AB - For epidemiologic studies that evaluate the relation between potential exposures to environmental chemicals and adverse outcomes, accurate assessments of exposures and health outcomes are needed. Three prospective cohort studies recently evaluated the relation between exposure, as assessed by biomonitoring, of pregnant women to organophosphorus pesticides and several birth outcomes. Here these three studies are compared in terms of the exposure scenarios and exposure assessments. The primary focus is on the exposure assessments, all of which employ biomonitoring but use different approaches, which may contribute to the different findings. These approaches and how they may contribute to different relations between exposure and birth outcomes are examined. PMID- 15811843 TI - Variation in organophosphate pesticide metabolites in urine of children living in agricultural communities. AB - Children of migrant farmworkers are at increased risk of exposure to organophosphate pesticides because of "carry-home" transport processes and residential location. Although this at-risk status is generally recognized, few available reports describe the extent of this exposure among agricultural communities. We quantified dialkyl phosphate (DAP) levels in serial samples of urine from 176 children, 2-6 years of age, in three Oregon communities hosting differing agricultural industries: pears, cherries, and fruit berries. Up to three spot samples of urine were collected from children at the beginning, mid point, and end of their parents' work seasons. The median levels of dimethylthiophosphate (DMTP), the most commonly detected metabolite, was significantly higher in urine samples from children in each of the three agricultural communities (17.5, 19.0, and 41.0 ng/mL) relative to a reference group of children who lived in an urban community and whose parents did not work in agriculture (6.5 ng/mL; Kruskal-Wallis, p < 0.001). After controlling for age, sex, and weight, the median level of DMTP in children in the pear community was 1.92 times higher than the level in children of the berry community [95% confidence interval (CI), 1.14-3.23] and 1.75 times higher than the level in children of the cherry community (95% CI, 0.95-3.23). We observed increasing levels of DMTP across the work season only within the berry community. Levels decreased in the cherry community and remained constant in the pear community. Substantial temporal variation within the children followed demonstrates the need for multiple urine samples to most accurately characterize longer term and/or cumulative exposure. The observed variability in urinary DAP levels, between communities and over time, could be attributed to the types and amounts of organophosphate pesticides used, the timing of applications and degradation of residues in the environment, work operations and hygiene practices, the proximity of housing to orchards and fields, or the movement of these working families. Additional studies of variation in pesticide exposure across agricultural regions are needed. PMID- 15811844 TI - Age-related differences in susceptibility to carcinogenesis. II. Approaches for application and uncertainty analyses for individual genetically acting carcinogens. AB - In an earlier report we developed a quantitative likelihood-based analysis of the differences in sensitivity of rodents to mutagenic carcinogens across three life stages (fetal, birth to weaning, and weaning to 60 days) relative to exposures in adult life. Here we draw implications for assessing human risks for full lifetime exposures, taking into account three types of uncertainties in making projections from the rodent data: uncertainty in the central estimates of the life-stage specific sensitivity factors estimated earlier, uncertainty from chemical-to chemical differences in life-stage-specific sensitivities for carcinogenesis, and uncertainty in the mapping of rodent life stages to human ages/exposure periods. Among the uncertainties analyzed, the mapping of rodent life stages to human ages/exposure periods is most important quantitatively (a range of several-fold in estimates of the duration of the human equivalent of the highest sensitivity "birth to weaning" period in rodents). The combined effects of these uncertainties are estimated with Monte Carlo analyses. Overall, the estimated population arithmetic mean risk from lifetime exposures at a constant milligrams per kilogram body weight level to a generic mutagenic carcinogen is about 2.8 fold larger than expected from adult-only exposure with 5-95% confidence limits of 1.5- to 6-fold. The mean estimates for the 0- to 2-year and 2- to 15-year periods are about 35-55% larger than the 10- and 3-fold sensitivity factor adjustments recently proposed by the U.S. Environmental Protection Agency. The present results are based on data for only nine chemicals, including five mutagens. Risk inferences will be altered as data become available for other chemicals. PMID- 15811845 TI - Results of a phase II trial of concurrent chemoradiation in the treatment of locally advanced carcinoma of uterine cervix: an experience from India. AB - OBJECTIVES: This prospective study was carried out to evaluate the results of chemo radiation (CRT) in the treatment of locally advanced carcinoma cervix and to compare with patients treated with radiotherapy (RT) alone. METHODS: Between 1996 and 2001, 57 patients of carcinoma cervix (bulky > 4 cm clinically, stage IIB/III) were treated with CRT. External radiation 46 Gy/23 fractions was followed by brachytherapy 35 Gy to Point A. Cisplatin was given in a dose of 50mg /week concurrently with external radiation for 5 cycles. A non-randomized comparison of CRT was done with the same subset of patients treated with RT alone (895 pts) during the same period. RESULTS: Overall survival (OS), disease free survival (DFS) and pelvic control with CRT at 5 years (45.6 %, 43.9 % and 57.9 %) were similar to RT alone arm (47.4 %, 45.5 % and 61.3 %). Compared to RT alone arm, (11.6 %) severe acute toxicities (grade 3 > or =, RTOG criteria) were significantly higher in CRT arm (31.6 %) (p < 0.001). More late rectal toxicities were encountered in CRT arm (22.8%) compared to RT alone (12.5 %) (p = 0.01), but the incidence of severe late rectal toxicities (> or = grade 3) were similar; 1.8 % in CRT versus 1.1 % in RT arm. CONCLUSIONS: With CRT, there were no significant differences in the outcomes and the incidence of severe late toxicities compared to RT alone arm. As the patients in Indian subcontinent usually present in late stages with poor general condition, CRT should be used judiciously with careful attention given to patient selection with escalation of doses of cisplatin to get the optimal benefit. PMID- 15811846 TI - [Regulation of telomeres length: making the telomeres accessible?]. AB - Under a normal state, the extremities of chromosomes, telomeres, are protected against undesired fusion events. Alterations of the telomere structure are associated with genetic instability, while erosion of the telomeric repeats, occurring at each cell division, provides a mechanism controlling the long-term proliferation of somatic cells. Although the structure and composition of the human telomerase enzyme are now well characterized, the protein partners regulating the stability and conformation of its DNA substrate, the telomeric end, are much less known. A functional link has been recently evidenced between proteins that bind the double-stranded telomere repeats and those recruited at the 3' G-rich telomeric overhang. This review presents an update on these telomeric factors controlling telomere lengthening and discuss the actual models proposed for its regulation. PMID- 15811847 TI - [Respiratory gated radiotherapy: the 4D radiotherapy]. AB - Respiration-gated radiotherapy offers a significant potential for improvement in the irradiation of tumor sites affected by respiratory motion such as lung, breast and liver tumors. An increased conformality of irradiation fields leading to decreased complications rates of organs at risk (lung, heart...) is expected. Respiratory gating is in line with the need for improved precision required by radiotherapy techniques such as 3D conformal radiotherapy or intensity modulated radiotherapy. Reduction of respiratory motion can be achieved by using either breath hold techniques or respiration synchronized gating techniques. Breath hold techniques can be achieved with active, in which airflow of the patient is temporarily blocked by a valve, or passive techniques, in which the patient voluntarily breath-hold. Synchronized gating techniques use external devices (CCD camera for the GEMS/Varian system tested at Curie Institute) to predict the phase of the respiration cycle while the patient breaths freely. A new strategy is currently developed: the 4D Respiration correlated CT. It consists of retrospectively reconstruct CT slices at different phases of the breathing cycle allowing to measure residual movements and to choose the optimal patient's breathing phase where tumor movements are lower. These techniques presently investigated in several medical centers worldwide. The first results are very promising. PMID- 15811848 TI - Drosophila lysyl oxidases Dmloxl-1 and Dmloxl-2 are differentially expressed and the active DmLOXL-1 influences gene expression and development. AB - Mammalian lysyl oxidase (LOX) is essential for the catalysis of lysyl-derived cross-links in fibrillar collagens and elastin in the extracellular matrix and has also been implicated in cell motility, differentiation, and tumor cell invasion. The active LOX has been shown to translocate to the nuclei of smooth muscle cells and regulate chromatin structure and transcription. It is difficult to interpret the role of the LOX protein as it is co-expressed with other members of the LOX amine oxidase family in most mammalian cells. To investigate the function of the LOX proteins, we have characterized the Drosophila lysyl oxidases Dmloxl-1 and Dmloxl-2. We present the gene, domain structure, and expression pattern of Dmloxl-1 and Dmloxl-2 during development. In early development, only Dmloxl-1 was expressed, which allowed functional studies. We have expressed Dmloxl-1 in S2 cells and determined that it is a catalytically active enzyme, inhibited by beta-amino-proprionitrile (BAPN), a specific LOX inhibitor. We localized DmLOXL-1 in the nuclei in embryos and in adult salivary gland cells in the nuclei, cytoplasm, and cell surface, using immunostaining and a DmLOXL-1 antibody. To address the biological function of Dmloxl-1, we raised larvae under BAPN inhibitory conditions and over-expressed Dmloxl-1 in transgenic Drosophila. DmLOXL-1 inhibition resulted in developmental delay and a shift in sex ratio; over-expression in the w(m4) variegating strain increased drosopterin production, demonstrating euchromatinization. Our previous data on the transcriptional down regulation of seven ribosomal genes and the glue gene under inhibitory conditions and the current results collectively support a nuclear role for Dmloxl-1 in euchromatinization and gene regulation. PMID- 15811849 TI - An amino-terminal motif functions as a second nuclear export sequence in BRCA1. AB - Mutations in the breast cancer susceptibility gene 1 (BRCA1) account for a substantial percentage of familial breast and ovarian cancers. Although BRCA1 is thought to function within the nucleus, it has also been located in the cytoplasm. In addition, BRCA1 accumulates in the nucleus of cells treated with leptomycin B, an inhibitor of chromosome region maintenance 1-mediated nuclear export, indicative of its active nuclear export via this pathway. The nuclear export signal in BRCA1 has been described as consisting of amino acid residues 81 99. However, a number of other tumor suppressors have multiple nuclear export sequences, and we sought to determine whether BRCA1 did also. Here, we report that BRCA1 contains a second nuclear export sequence that comprises amino acid residues 22-30. By use of the human immunodeficiency virus-1 Rev complementation assay, this sequence was shown to confer export capability to an export-defective Rev fusion protein. The level of export activity was comparable with that of residues 81-99 comprising the previously reported nuclear export sequence in BRCA1. Mutation of leucine 28 to an alanine reduced nuclear export by approximately 75%. In MCF-7 cells stably transfected with a BRCA1 cDNA containing mutations in this novel sequence or the previously reported export sequence, BRCA1 accumulated in the nucleus. These data imply that BRCA1 contains at least two leucine-dependent nuclear export sequences. PMID- 15811850 TI - Substrate specificity of tyrosyl-DNA phosphodiesterase I (Tdp1). AB - Tyrosyl-DNA phosphodiesterase I (Tdp1) hydrolyzes 3'-phosphotyrosyl bonds to generate 3'-phosphate DNA and tyrosine in vitro. Tdp1 is involved in the repair of DNA lesions created by topoisomerase I, although the in vivo substrate is not known. Here we study the kinetic and binding properties of human Tdp1 (hTdp1) to identify appropriate 3'-phosphotyrosyl DNA substrates. Genetic studies argue that Tdp1 is involved in double and single strand break repair pathways; however, x ray crystal structures suggest that Tdp1 can only bind single strand DNA. Separate kinetic and binding experiments show that hTdp1 has a preference for single-stranded and blunt-ended duplex substrates over nicked and tailed duplex substrate conformations. Based on these results, we present a new model to explain Tdp1/DNA binding properties. These results suggest that Tdp1 only acts upon double strand breaks in vivo, and the roles of Tdp1 in yeast and mammalian cells are discussed. PMID- 15811851 TI - Comparative functional study of the viral telomerase RNA based on natural mutations. AB - Telomerase activity is present in most malignant tumors and provides a mechanism for the unlimited potential for division of neoplastic cells. We previously characterized the first identified viral telomerase RNA (vTR) encoded by the Marek's disease virus (MDV) (Fragnet, L., Blasco, M. A., Klapper, W., and Rasschaert, D. (2003) J. Virol. 77, 5985-5996). This avian herpesvirus induces T lymphomas. We demonstrated that the vTR subunit of the oncogenic MDV-RB1B strain is functional and would be more efficient than its chicken counterpart, cTR, which is 88% homologous. We take advantage of the functionality of those natural mutant TRs to investigate the involvement of the mutations of vTR on its efficiency in a heterologous murine cell system and in a homologous in vitro system using the recombinant chicken telomerase reverse transcriptase. The P2 helix of the pseudoknot seems to be more stable in vTR than in cTR, and this may account for the higher activity of vTR than cTR. Moreover, the five adenines just upstream from the P3 helix of vTR may also play an important role in its efficiency. We also established that the substitution of a single nucleotide at the 3'-extremity of the H-box of the vaccine MDV-Rispens strain vTR resulted in a lack of its accumulation within the cell, especially in the nucleus, correlated with a decrease in telomerase activity. PMID- 15811852 TI - Coactivators and corepressors of NF-kappaB in IkappaB alpha gene promoter. AB - In this study, we investigated recruitment of coactivators (SRC-1, SRC-2, and SRC 3) and corepressors (HDAC1, HDAC2, HDAC3, SMRT, and NCoR) to the IkappaB alpha gene promoter after NF-kappaB activation by tumor necrosis factor-alpha. Our data from chromatin immunoprecipitation assay suggest that coactivators and corepressors are simultaneously recruited to the promoter, and their binding to the promoter DNA is oscillated in HEK293 cells. SRC-1, SRC-2, and SRC-3 all enhanced IkappaB alpha transcription. However, the interaction of each coactivator with the promoter exhibited different patterns. After tumor necrosis factor-alpha treatment, SRC-1 signal was increased gradually, but SRC-2 signal was reduced immediately, suggesting replacement of SRC-2 by SRC-1. SRC-3 signal was increased at 30 min, reduced at 60 min, and then increased again at 120 min, suggesting an oscillation of SRC-3. The corepressors were recruited to the promoter together with the coactivators. The binding pattern suggests that the corepressor proteins formed two types of corepressor complexes, SMRT-HDAC1 and NCoR-HDAC3. The two complexes exhibited a switch at 30 and 60 min. The functions of cofactors were confirmed by gene overexpression and RNA interference-mediated gene knockdown. These data suggest that gene transactivation by the transcription factor NF-kappaB is subject to the regulation of a dynamic balance between the coactivators and corepressors. This model may represent a mechanism for integration of extracellular signals into a precise control of gene transcription. PMID- 15811853 TI - Smad7 abrogates transforming growth factor-beta1-mediated growth inhibition in COLO-357 cells through functional inactivation of the retinoblastoma protein. AB - Smad7 is overexpressed in 50% of human pancreatic cancers. COLO-357 pancreatic cancer cells engineered to overexpress Smad7 are resistant to the actions of transforming growth factor-beta1 (TGF-beta1) with respect to growth inhibition and cisplatin-induced apoptosis but not with respect to modulation of gene expression. To delineate the mechanisms underlying these divergent consequences of Smad7 overexpression, we studied the effects of Smad7 on TGF-beta1-dependent signaling pathways and cell cycle regulating proteins. TGF-beta1 induced the phosphorylation of MAPK, p38 MAPK, and AKT2 irrespective of the levels of Smad7, and inhibitors of these pathways did not alter TGF-beta1 actions on cell growth. By contrast, Smad7 overexpression interfered with TGF-beta1-mediated attenuation of cyclin A and B levels, inhibition of cdc2 dephosphorylation and CDK2 inactivation, up-regulation of p27, and the maintenance of the retinoblastoma protein (RB) in a hypophosphorylated state. Smad7 also suppressed TGF-beta1 mediated inhibition of E2F activity but did not alter TGF-beta1-mediated phosphorylation of Smad2, the nuclear translocation of Smad2/3/4, or DNA binding of the Smad2/3/4 complex. Although Smad7 did not associate with the type I TGF beta receptor (TbetaRI), SB-431542, an inhibitor of the kinase activity of this receptor, blocked TGF-beta1-mediated effects on Smad-2 phosphorylation. These findings point toward a novel paradigm whereby Smad7 acts to functionally inactivate RB and de-repress E2F without blocking the activation of TbetaRI and the nuclear translocation of Smad2/3, thereby allowing for TGF-beta1 to exert effects in a cancer cell that is resistant to TGF-beta1-mediated growth inhibition. PMID- 15811854 TI - Further characterization of human DNA polymerase delta interacting protein 38. AB - Polymerase delta interacting protein 38 (PDIP38) was identified as a human DNA polymerase (pol) delta interacting protein through a direct interaction with p50, the small subunit of human pol delta. PDIP38 was also found to interact with proliferating cell nuclear antigen, which suggested that it might play a role in vivo in the processes of DNA replication and DNA repair in the nucleus. In order to characterize further this novel protein, we have examined its subcellular localization by the use of immunochemical and cellular fractionation techniques. These studies show that PDIP38 is a novel mitochondrial protein and is localized mainly to the mitochondria. PDIP38 was shown to possess a functional mitochondrial targeting sequence that is located within the first 35 N-terminal amino acid residues. The mature PDIP38 protein is about 50 amino acid residues smaller than the full-length precursor PDIP38 protein, consistent with it being processed by cleavage of the mitochondrial targeting sequence during entry into the mitochondria. His-tagged mature PDIP38 inhibited pol delta activity in vitro and interacted with human papillomavirus 16 E7 oncoprotein, suggesting that PDIP38 might play a role in the pol delta-mediated viral DNA replication. Although the localization of PDIP38 to the mitochondria suggests that it serves functions within the mitochondria, we cannot eliminate the possibility that it may be involved in pol delta-mediated DNA replication or DNA repair under certain conditions such as viral infection. PMID- 15811855 TI - Cytokines induce nitric oxide-mediated mtDNA damage and apoptosis in oligodendrocytes. Protective role of targeting 8-oxoguanine glycosylase to mitochondria. AB - Nitric oxide (NO) that is produced by inducible NO synthase (iNOS) in glial cells is thought to contribute significantly to the pathogenesis of multiple sclerosis. Oligodendrocytes can be stimulated to express iNOS by inflammatory cytokines, which are known to accumulate in the multiple sclerotic brain. The potentially pathological levels of NO produced under these circumstances can target a wide spectrum of intracellular components. We hypothesized that one of the critical targets for damage that leads to disease is mtDNA. In this study, we found that cytokines, in particular a combination of tumor necrosis factor-alpha (50 ng/ml) and IFNgamma (25 ng/ml), cause elevated NO production in primary cultures of rat oligodendrocytes. Western blot analysis revealed a strong enhancement of iNOS expression 48 h after cytokine treatment. Within the same time period, NO mediated mtDNA damage was shown by Southern blot analysis and by ligation mediated PCR. Targeting the DNA repair enzyme human 8-oxoguanine DNA glycosylase (hOGG1) to the mitochondria of oligodendrocytes had a protective effect against this cytokine-mediated mtDNA damage. Moreover, it was shown that mitochondrial transport sequence hOGG1-transfected oligodendrocytes had fewer apoptotic cells compared with cells containing vector only following treatment with the cytokines. Subsequent experiments revealed that targeting hOGG1 to mitochondria reduces the activation of caspase-9, showing that this recombinant protein works to reduce apoptosis that is occurring through a mitochondria-based pathway. PMID- 15811856 TI - Dynamic transcriptional regulatory complexes, including E2F4, p107, p130, and Sp1, control fibroblast growth factor receptor 1 gene expression during myogenesis. AB - Developmentally controlled transcriptional regulation of myogenic cell proliferation and differentiation via expression of the fibroblast growth factor receptor 1 (FGFR1) gene is positively regulated by Sp1 and negatively regulated by E2F4-based transcriptional complexes. We report that p107 and p130 formed transcriptional complexes with E2F4 on the FGFR1 promoter and repressed FGFR1 gene transcription in myogenic cells. However, in Drosophila melanogaster SL2 cells, only p107 was able to repress Sp1-mediated transactivation of the FGFR1 promoter. Gel shift assays using transfected myoblast nuclear extracts showed that ectopic p107 reduced Sp1 occupancy of the proximal Sp binding site of the FGFR1 promoter, and coimmunoprecipitation studies indicated that Sp1 interacts with p107 but not with p130. Gel shift assays also demonstrated that Sp1 interacted with p107 in E2F4-p107 transcriptional complexes in myoblasts. The nature of the repressor transcriptional complex was altered in differentiated muscle fibers by the relative loss of the E2F4-p107-Sp1 transcription complex and replacement by the repressor E2F4-p130 complex. These findings demonstrate that activation and repression of FGFR1 gene transcription is governed by interplay between Sp1, p107, p130, and E2F4 in distinct transcriptional complexes during skeletal muscle development. PMID- 15811857 TI - Collagen phagocytosis by fibroblasts is regulated by decorin. AB - Decorin is a small, leucine-rich proteoglycan that binds to collagen and regulates fibrillogenesis. We hypothesized that decorin binding to collagen inhibits phagocytosis of collagen fibrils. To determine the effects of decorin on collagen degradation, we analyzed phagocytosis of collagen and collagen/decorin coated fluorescent beads by Rat-2 and gingival fibroblasts. Collagen beads bound to gingival cells by alpha2beta1 integrins. Binding and internalization of decorin/collagen-coated beads decreased dose-dependently with increasing decorin concentration (p < 0.001). Inhibition of binding was sustained over 5 h (p < 0.001) and was attributed to interactions between decorin and collagen and not to decorin-collagen receptor interactions. Both the non-glycosylated decorin core protein and the thermally denatured decorin significantly inhibited collagen bead binding (approximately 50 and 89%, respectively; p < 0.05). Mimetic peptides corresponding to leucine-rich repeats 1-3, encompassed by a collagen-binding approximately 11-kDa cyanogen bromide fragment of decorin and leucine-rich repeats 4 and 5, previously shown to bind to collagen, were tested for their ability to inhibit collagen bead binding. Although the synthetic peptide 3 alone exhibited saturable binding to collagen, neither peptides 3 nor 1 and 2 markedly inhibited phagocytosis. Leucine-rich repeat 3 bound to a triple helical peptide containing the alpha2 integrin-binding site of collagen. When collagen beads were co-incubated with peptides 3 and 4, inhibition of collagen phagocytosis (55%) was equivalent to intact native/recombinant core protein. Thus a novel collagen binding domain in decorin acts cooperatively with leucine-rich repeat 4 to mask the alpha2beta1 integrin-binding site on collagen, an important sequence for the phagocytosis of collagen fibrils. PMID- 15811858 TI - Analysis of the interaction between the Saccharomyces cerevisiae MSH2-MSH6 and MLH1-PMS1 complexes with DNA using a reversible DNA end-blocking system. AB - The Lac repressor-operator interaction was used as a reversible DNA end-blocking system in conjunction with an IAsys biosensor instrument (Thermo Affinity Sensors), which detects total internal reflectance and allows monitoring of binding and dissociation in real time, in order to develop a system for studying the ability of mismatch repair proteins to move along the DNA. The MSH2-MSH6 complex bound to a mispaired base was found to be converted by ATP binding to a form that showed rapid sliding along the DNA and dissociation via the DNA ends and also showed slow, direct dissociation from the DNA. In contrast, the MSH2 MSH6 complex bound to a base pair containing DNA only showed direct dissociation from the DNA. The MLH1-PMS1 complex formed both mispair-dependent and mispair independent ternary complexes with the MSH2-MSH6 complex on DNA. The mispair independent ternary complexes were formed most efficiently on DNA molecules with free ends under conditions where ATP hydrolysis did not occur, and only exhibited direct dissociation from the DNA. The mispair-dependent ternary complexes were formed in the highest yield on DNA molecules with blocked ends, required ATP and magnesium for formation, and showed both dissociation via the DNA ends and direct dissociation from the DNA. PMID- 15811859 TI - Caenorhabditis elegans geminin homologue participates in cell cycle regulation and germ line development. AB - Cdt1 is an essential component for the assembly of a pre-replicative complex. Cdt1 activity is inhibited by geminin, which also participates in neural development and embryonic differentiation in many eukaryotes. Although Cdt1 homologues have been identified in organisms ranging from yeast to human, geminin homologues had not been described for Caenorhabditis elegans and fungi. Here, we identify the C. elegans geminin, GMN-1. Biochemical analysis reveals that GMN-1 associates with C. elegans CDT-1, the Hox protein NOB-1, and the Six protein CEH 32. GMN-1 inhibits not only the interaction between mouse Cdt1 and Mcm6 but also licensing activity in Xenopus egg extracts. RNA interference-mediated reduction of GMN-1 is associated with enlarged germ nuclei with aberrant nucleolar morphology, severely impaired gametogenesis, and chromosome bridging in intestinal cells. We conclude that the Cdt1-geminin system is conserved throughout metazoans and that geminin has evolved in these taxa to regulate proliferation and differentiation by directly interacting with Cdt1 and homeobox proteins. PMID- 15811860 TI - It's the premiums, stupid: projections of the uninsured through 2013. AB - Increases in the cost of health care from 1979 to 1999 accounted for the decline in health insurance coverage that occurred during that time period, as our earlier work demonstrated. Here we examine whether the model we presented adequately accounts for the observed changes in health insurance coverage from 1999 through 2002, and we show that the model accurately predicted the increase in uninsured people during that time period. Using the model and projections for national health spending, we project that the number of nonelderly uninsured Americans will grow from forty-five million in 2003 to fifty-six million by 2013. PMID- 15811861 TI - Cardiology patient page. Peripheral arterial disease. PMID- 15811862 TI - Images in cardiovascular medicine. Torsade de pointes in a patient with Wolff Parkinson-White syndrome. PMID- 15811863 TI - Letter regarding article by Okoshi et al, "neuregulins regulate cardiac parasympathetic activity: muscarinic modulation of {beta}-adrenergic activity in myocytes from mice with neuregulin-1 gene deletion". PMID- 15811864 TI - Letter regarding article by Hlatky et al, "medical costs and quality of life 10 to 12 years after randomization to angioplasty or bypass surgery for multivessel coronary artery disease". PMID- 15811865 TI - Childhood infection and endothelial dysfunction: a potential link in atherosclerosis? PMID- 15811866 TI - A new "sunshine" in the vasculature? PMID- 15811867 TI - Phosphodiesterase-5 inhibition with sildenafil attenuates cardiomyocyte apoptosis and left ventricular dysfunction in a chronic model of doxorubicin cardiotoxicity. AB - BACKGROUND: Sildenafil, a phosphodiesterase-5 inhibitor, induces cardioprotection against ischemia/reperfusion injury via opening of mitochondrial K(ATP) channels. It is unclear whether sildenafil would provide similar protection from doxorubicin-induced cardiotoxicity. METHODS AND RESULTS: Male ICR mice were randomized to 1 of 4 treatments: saline, sildenafil, doxorubicin (5 mg/kg IP), and sildenafil (0.7 mg/kg IP) plus doxorubicin (n=6 per group). Apoptosis was assessed with the use of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and in situ oligo ligation methods. Desmin distribution was determined via immunofluorescence. Bcl-2 expression was analyzed by Western blot. Left ventricular function was assessed by measuring developed pressure and rate pressure product in Langendorff mode. ECG changes indicative of doxorubicin cardiotoxicity were also measured. For in vitro studies, adult ventricular cardiomyocytes were exposed to doxorubicin (1 micromol/L), sildenafil (1 micromol/L) with or without N(G)-nitro-L-arginine methyl ester (L-NAME) (100 micromol/L), or 5-hydroxydecanoate (100 micromol/L) 1 hour before doxorubicin and incubated for 18 hours. Doxorubicin-treated mice demonstrated increased apoptosis and desmin disruption, which was attenuated in the sildenafil+doxorubicin group. Bcl-2 was decreased in the doxorubicin group but was maintained at basal levels in the sildenafil+doxorubicin group. Left ventricular developed pressure and rate pressure product were significantly depressed in the doxorubicin group but were attenuated in the sildenafil+doxorubicin group. ST interval was significantly increased in the doxorubicin group over 8 weeks. In the sildenafil+doxorubicin group, ST interval remained unchanged from baseline. Doxorubicin caused a significant increase in apoptosis, caspase-3 activation, and disruption of mitochondrial membrane potential in vitro. In contrast, sildenafil significantly protected against doxorubicin cardiotoxicity; however, this protection was abolished by both L-NAME and 5-hydroxydecanoate. CONCLUSIONS: Prophylactic treatment with sildenafil prevented apoptosis and left ventricular dysfunction in a chronic model of doxorubicin-induced cardiomyopathy. PMID- 15811868 TI - Gender differences in outcomes after primary angioplasty versus primary stenting with and without abciximab for acute myocardial infarction: results of the Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trial. AB - BACKGROUND: Women with acute myocardial infarction (AMI) undergoing primary angioplasty have higher rates of morbidity and mortality than do men. Whether contemporary interventional treatment strategies have improved outcomes for women compared with men is unknown. METHODS AND RESULTS: In the CADILLAC trial, 2082 patients (27% women) with AMI within 12 hours of symptom onset were randomized to balloon angioplasty (PTCA; n=518), PTCA+abciximab (n=528), stenting (n=512), and stenting+abciximab (n=524). As compared with men, women had a lower body surface area; had a greater prevalence of diabetes, hypertension, and hyperlipidemia; experienced significant delays to treatment; and had better baseline and final TIMI grade 3 flows. Unadjusted 1-year event rates were higher for women, including death (7.6% versus 3.0%, P<0.001), ischemic target-vessel revascularization (TVR; 16.7% versus 12.1%, P=0.006), and major adverse cardiac events (MACE; 23.9% versus 15.3%, P<0.001). Female gender was an independent predictor of MACE and bleeding complications, although comorbid risk factors and body surface area but not gender predicted 1-year death. For women, primary stenting resulted in a reduction in 1-year MACE from 28.1% to 19.1% (P=0.01) and in ischemic TVR from 20.4% to 10.8% (P=0.002) compared with PTCA. The addition of abciximab to primary stenting significantly reduced the 30-day ischemic TVR without increasing bleeding or stroke rates. CONCLUSIONS: The higher mortality rate in women compared with men after interventional treatment for AMI may be explained by differences in body size and clinical risk factors, although female gender remains an important independent determinant of overall adverse outcomes. For women in the CADILLAC trial, the addition of abciximab reduced 30-day TVR without increasing bleeding risk, and primary stenting reduced 1-year TVR and MACE rates compared with PTCA. PMID- 15811869 TI - The US Food and Drug Administration Cardiorenal Advisory Panel and the drug approval process. PMID- 15811870 TI - American College of Cardiology and American Heart Association methodology for the selection and creation of performance measures for quantifying the quality of cardiovascular care. AB - The ability to quantify the quality of cardiovascular care critically depends on the translation of recommendations for high-quality care into the measurement of that care. As payers and regulatory agencies increasingly seek to quantify healthcare quality, the implications of the measurement process on practicing physicians are likely to grow. This statement describes the methodology by which the American College of Cardiology and the American Heart Association approach creating performance measures and devising techniques for quantifying those aspects of care that directly reflect the quality of cardiovascular care. Methods for defining target populations, identifying dimensions of care, synthesizing the literature, and operationalizing the process of selecting measures are proposed. It is hoped that new sets of measures will be created through the implementation of this approach, and consequently, through the use of such measurement sets in the context of quality improvement efforts, the quality of cardiovascular care will improve. PMID- 15811874 TI - Gonadotropin regulation of glutamate cysteine ligase catalytic and modifier subunit expression in rat ovary is subunit and follicle stage specific. AB - We have observed that levels of the antioxidant glutathione (GSH) and protein levels of the catalytic and modifier subunits of the rate-limiting enzyme in GSH synthesis, GCLc and GCLm, increase in immature rat ovaries after treatment with gonadotropin. The goals of the present studies were to delineate the time course and intraovarian localization of changes in GSH and GCL after pregnant mare's serum gonadotropin (PMSG) and after an ovulatory gonadotropin stimulus. Twenty four hours after PMSG, there was a shift from predominantly granulosa cell expression of gclm mRNA, and to a lesser extent gclc, to predominantly theca cell expression. GCLc immunostaining increased in granulosa and theca cells and in interstitial cells. Next, prepubertal female rats were primed with PMSG, followed 48 h later by 10 IU of hCG. GCLm protein and mRNA levels increased dramatically from 0 to 4 h after hCG and then declined rapidly. There was minimal change in GCLc. The increase in gclm mRNA expression was localized mainly to granulosa and theca cells of preovulatory follicles. To verify that GCL responds similarly to an endogenous preovulatory gonadotropin surge, we quantified ovarian GCL mRNA levels during the periovulatory period in adult rats. gclm mRNA levels increased after the gonadotropin surge on proestrus and then declined rapidly. Finally, we assessed the effects of gonadotropin on ovarian GCL enzymatic activity. GCL enzymatic activity increased significantly at 48 h after PMSG injection and did not increase further after hCG. These results demonstrate that gonadotropins regulate follicular GCL expression in a follicle stage-dependent manner and in a GCL subunit-dependent manner. PMID- 15811875 TI - Thapsigargin-sensitive cationic current leads to membrane depolarization, calcium entry, and insulin secretion in rat pancreatic beta-cells. AB - Glucose-induced insulin secretion by pancreatic beta-cells depends on membrane depolarization and [Ca2+]i increase. We correlated voltage- and current-clamp recordings, [Ca2+]i measurements, and insulin reverse hemolytic plaque assay to analyze the activity of a thapsigargin-sensitive cationic channel that can be important for membrane depolarization in single rat pancreatic beta-cells. We demonstrate the presence of a thapsigargin-sensitive cationic current, which is mainly carried by Na+. Moreover, in basal glucose concentration (5.6 mM), thapsigargin depolarizes the plasma membrane, producing electrical activity and increasing [Ca2+]i. The latter is prevented by nifedipine, indicating that Ca2+ enters the cell through L-type Ca2+ channels, which are activated by membrane depolarization. Thapsigargin also increased insulin secretion by increasing the percentage of cells secreting insulin and amplifying hormone secretion by individual beta-cells. Nifedipine blocked the increase completely in 5.6 mM glucose and partially in 15.6 mM glucose. We conclude that thapsigargin potentiates a cationic current that depolarizes the cell membrane. This, in turn, increases Ca2+ entry through L-type Ca2+ channels promoting insulin secretion. PMID- 15811876 TI - Secretory dynamics of leptin in adolescent girls with anorexia nervosa and healthy adolescents. AB - Leptin, an adipocytokine that suppresses appetite and may regulate neuroendocrine pathways, is low in undernourished states like anorexia nervosa (AN). Although leptin exhibits pulsatility, secretory characteristics have not been well described in adolescents and in AN, and the contribution of hypoleptinemia to increased growth hormone (GH) and cortisol in AN has not been explored. We hypothesized that hypoleptinemia in AN reflects decreased basal and pulsatile secretion and may predict increased GH and cortisol levels. Sampling for leptin, GH, cortisol, and ghrelin was performed every 30 min (from 2000 to 0800) in 23 AN and 21 controls 12-18 yr old, and data were analyzed using Cluster and deconvolution methods. Estradiol, thyroid hormones, and body composition were measured. AN girls had lower pulsatile and total leptin secretion than controls (P < 0.0001) subsequent to decreased burst mass (P < 0.0001) and basal secretion (P = 0.02). Nutritional markers predicted leptin characteristics. In a regression model including BMI, body fat, and ghrelin, leptin independently predicted GH burst interval and frequency. Valley leptin contributed to 56% of the variability in GH burst interval, and basal leptin and fasting ghrelin contributed to 42% of variability in burst frequency. Pulsatile leptin independently predicted urine free cortisol/creatinine (15% of variability). Valley leptin predicted cortisol half-life (22% of variability). Leptin predicted estradiol and thyroid hormone levels. In conclusion, hypoleptinemia in AN is subsequent to decreased basal and pulsatile secretion and nutritionally regulated. Leptin predicts GH and cortisol parameters and with ghrelin predicts GH burst frequency. Low leptin and high ghrelin may be dual stimuli for high GH concentrations in undernutrition. PMID- 15811877 TI - Physiological activation of hypoxia inducible factor-1 in human skeletal muscle. AB - The human hypoxia inducible factor 1 (HIF-1) system is activated under various pathological conditions, yet less is known about its physiological regulation in healthy human tissue. We have studied the effect of exercise on the activation of HIF-1 in human skeletal muscle. Employing a model where oxygen consumption increases and oxygen tension can be manipulated, nine healthy male subjects performed 45 min of one-legged knee-extension exercise. Biopsies were taken before, directly after, and 30, 120, and 360 min after exercise. Exercise led to elevated HIF-1alpha protein levels and a more prevalent nuclear staining of HIF 1alpha. Interestingly, a concurrent decrease in von Hippel-Lindau tumor suppressor protein (VHL) levels was detected in some subjects. Moreover, exercise induced an increase in the DNA binding activity of HIF-1alpha. Characterization of gene expression by real-time PCR demonstrated that the HIF-1 target genes VEGF and EPO were activated. VEGF mRNA was further increased when blood flow to the exercising leg was restricted. In conclusion, these data clearly demonstrate that physical activity induces the HIF-1-mediated signaling pathway in human skeletal muscle, providing the first evidence that human HIF-1alpha can be activated during physiologically relevant conditions. PMID- 15811878 TI - p53 is a suppressor of inflammatory response in mice. AB - Chronic inflammation is known to promote cancer, suggesting that negative regulation of inflammation is likely to be tumor suppressive. We found that p53 is a general inhibitor of inflammation that acts as an antagonist of nuclear factor kappaB (NFkappaB). We first observed striking similarities in global gene expression profiles in human prostate cancer cells LNCaP transduced with p53 inhibitory genetic element or treated with TNF, suggesting that p53 inhibits transcription of TNF-inducible genes that are largely regulated by NFkappaB. Consistently, ectopically expressed p53 acts as an inhibitor of transcription of NFkappaB-dependent promoters. Furthermore, suppression of inflammatory response by p53 was observed in vivo in mice by comparing wild-type and p53 null animals at molecular (inhibition of transcription of genes encoding cytokines and chemokines, reducing accumulation of reactive oxygen species and protein oxidation products), cellular (activation of macrophages and neutrophil clearance) and organismal (high levels of metabolic markers of inflammation in tissues of p53-deficient mice and their hypersensitivity to LPS) levels. These observations indicate that p53, acting through suppression of NFkappaB, plays the role of a general "buffer" of innate immune response in vivo that is well consistent with its tumor suppressor function and frequent constitutive activation of NFkappaB in tumors. PMID- 15811879 TI - Arginase and polyamine synthesis are key factors in the regulation of experimental leishmaniasis in vivo. AB - Arginase 1, an enzyme induced by Th2 cytokines, is a hallmark of alternatively activated macrophages and is responsible for the hydrolysis of L-arginine into ornithine, the building block for the production of polyamines. Upregulation of arginase 1 has been observed in a variety of diseases, but the mechanisms by which arginase contributes to pathology are not well understood. We reveal here a unique role for arginase 1 in the pathogenesis of nonhealing leishmaniasis, a prototype Th2 disease, and demonstrate that the activity of this enzyme promotes pathology and uncontrolled growth of Leishmania parasites in vivo. Inhibition of arginase activity during the course of infection has a clear therapeutic effect, as evidenced by markedly reduced pathology and efficient control of parasite replication. Despite the clear amelioration of the disease, this treatment does not alter the Th2 response. To address the underlying mechanisms, the arginase induced L-arginine catabolism was investigated and the results demonstrate that arginase regulates parasite growth directly by affecting the polyamine synthesis in macrophages. PMID- 15811880 TI - Limitations of "pragmatic" trials. PMID- 15811882 TI - Scimitar syndrome (congenital pulmonary venolobar syndrome). PMID- 15811883 TI - Contemporary management of acute coronary syndrome. AB - This review focuses on the modern management of the non-ST elevation acute coronary syndromes (unstable angina and non-ST elevation myocardial infarction). Patients with these syndromes are at varying degrees of risk of (re)infarction and death. This risk can be reliably predicted by clinical, electrocardiographic, and biochemical markers. Aspirin, clopidogrel, heparin (unfractionated or low molecular weight), and anti-ischaemic drugs should be offered to all patients, irrespective of the predicted level of risk. Patients at high risk should also receive a glycoprotein IIb/IIIa receptor inhibitor and should undergo early coronary arteriography with a view to percutaneous or surgical revascularisation. Lower risk patients should undergo non-invasive testing. When inducible myocardial ischaemia is exhibited coronary arteriography should follow. When non invasive testing is negative, a conservative management strategy is safe. PMID- 15811881 TI - Acute poisoning: understanding 90% of cases in a nutshell. AB - The acutely poisoned patient remains a common problem facing doctors working in acute medicine in the United Kingdom and worldwide. This review examines the initial management of the acutely poisoned patient. Aspects of general management are reviewed including immediate interventions, investigations, gastrointestinal decontamination techniques, use of antidotes, methods to increase poison elimination, and psychological assessment. More common and serious poisonings caused by paracetamol, salicylates, opioids, tricyclic antidepressants, selective serotonin reuptake inhibitors, benzodiazepines, non-steroidal anti-inflammatory drugs, and cocaine are discussed in detail. Specific aspects of common paediatric poisonings are reviewed. PMID- 15811884 TI - Non-steroidal anti-inflammatory drugs and colorectal cancer prevention. AB - Colorectal cancer is the second leading cause of cancer deaths in the United States. Currently, the most effective strategy available for colon cancer prevention is endoscopic screening, with polypectomy or surgical resection for advanced lesions. This intervention carries with it many concerns regarding cost, patient acceptance, and the growing burden of surveillance colonoscopies for patients with polyps. Further improvements in the understanding of the multistep model of colorectal carcinogenesis will probably lead to the development of other primary and secondary prevention strategies. Data obtained from animal and epidemiological studies and most recently from randomised, placebo controlled trials, suggest that non-steroidal anti-inflammatory drugs may prove effective chemopreventive agents in different groups of people, from patients with familial adenomatous polyposis to those with sporadic adenomas. PMID- 15811885 TI - Substance misuse in the older population. AB - Substance misuse among the older population is largely overlooked and underreported. Many factors contribute to this, not least the fact that presentation may be atypical and hence easily missed by the medical practitioner. There may be many clues to its existence, provided the physician remains alert to these. Despite this it is quite comforting to know that once identified, the evidence to date suggests that older people may respond at least as well as younger people to treatment. PMID- 15811886 TI - Autonomic dysreflexia: a medical emergency. AB - Autonomic dysreflexia is an important clinical diagnosis that requires prompt treatment to avoid devastating complications. The condition may present itself to all members of medical and surgical specialties, who may not be accustomed to treating it. It is the clinician's responsibility to have a basic understanding of the pathophysiology of the condition and the simple steps required to treat it. PMID- 15811887 TI - Angiogenesis: a curse or cure? AB - Angiogenesis, the growth of new blood vessels is essential during fetal development, female reproductive cycle, and tissue repair. In contrast, uncontrolled angiogenesis promotes the neoplastic disease and retinopathies, while inadequate angiogenesis can lead to coronary artery disease. A balance between pro-angiogenic and antiangiogenic growth factors and cytokines tightly controls angiogenesis. Considerable progress has been made in identifying these molecular components to develop angiogenesis based treatments. One of the most specific and critical regulators of angiogenesis is vascular endothelial growth factor (VEGF), which regulates endothelial proliferation, permeability, and survival. Several VEGF based treatments including anti-VEGF and anti-VEGF receptor antibodies/agents are in clinical trials along with several other antiangiogenic treatments. While bevacizumab (anti-VEGF antibody) has been approved for clinical use in colorectal cancer, the side effects of antiangiogenic treatment still remain a challenge. The pros and cons of angiogenesis based treatment are discussed. PMID- 15811888 TI - Role of surgery in the management of the adult patient with coarctation of the aorta. AB - Adult patients with coarctation of the aorta have a range of clinical presentations. These include the presence of additional cardiovascular anomalies (predominantly aortic valve abnormality) and presentation with complications after coarctation repair in childhood (such as recurrent coarctation or aneurysm formation). Developments in endovascular technology over the past decade may potentially reduce the morbidity from open surgical repair. However, some cases are unsuitable for endovascular repair and open surgical techniques continue to play a part in the management of these patients. The number of adult patients with coarctation is very small and a trial comparing surgical and endovascular repair techniques would be difficult to arrange. A multidisciplinary team (consisting of cardiothoracic surgeon, interventional radiologist, cardiologist, and anaesthetist) is best equipped to manage the unique and complex problems that affect these patients. The current surgical options and factors governing the choice of approach are reviewed. PMID- 15811889 TI - Darwin's illness revealed. AB - After returning from the Beagle in 1836, Charles Darwin suffered for over 40 years from long bouts of vomiting, gut pain, headaches, severe tiredness, skin problems, and depression. Twenty doctors failed to treat him. Many books and papers have explained Darwin's mystery illness as organic or psychosomatic, including arsenic poisoning, Chagas' disease, multiple allergy, hypochondria, or bereavement syndrome. None stand up to full scrutiny. His medical history shows he had an organic problem, exacerbated by depression. Here we show that all Darwin's symptoms match systemic lactose intolerance. Vomiting and gut problems showed up two to three hours after a meal, the time it takes for lactose to reach the large intestine. His family history shows a major inherited component, as with genetically predisposed hypolactasia. Darwin only got better when, by chance, he stopped taking milk and cream. Darwin's illness highlights something else he missed--the importance of lactose in mammalian and human evolution. PMID- 15811890 TI - Prospective study of acute gastrointestinal bleeding attributable to anti inflammatory drug ingestion in the Yorkshire region of the United Kingdom. AB - OBJECTIVE: To assess the general use of all non-steroidal anti-inflammatory drugs (NSAID) and their relation to upper gastrointestinal bleeding in view of National Institute for Clinical Excellence guidelines published in July 2001 in the UK. METHODS: Cross sectional study on all patients who were referred for endoscopy for suspected upper gastrointestinal bleeding in six hospitals in Yorkshire region of the UK. RESULTS: One hundred and sixty three patients presented for endoscopy for suspected upper gastrointestinal bleeding, 43 patients were taking at least one ulcerogenic drug, and 120 were not. The mean age difference between these two groups was eight years (p<0.01). The absolute difference between the proportion of patients with peptic ulcer disease/erosion (PUD) in NSAID with/without aspirin group and no ulcerogenic drug group was 31% (p = 0.02). The difference between the proportion of PUD in cyclo-oxygenase 2 with/without aspirin group and no ulcerogenic drug group was 30% (p = 0.1). The overall 30 days mortality rate was 14.1%. CONCLUSIONS: Elderly patients are being inappropriately prescribed conventional NSAIDs. NSAIDs with or without aspirin use are still associated with a significant risk of upper gastrointestinal bleeding in the era of cyclo-oxygenase 2 selective agents. Substitution with cyclo-oxygenase 2 selective NSAIDs is not without risk of upper gastrointestinal bleeding. PMID- 15811891 TI - Four gland parathyroidectomy without reimplantation in patients with chronic renal failure. AB - BACKGROUND: The optimal surgical management of patients in end stage chronic renal failure with secondary hyperparathyroidism is controversial. One approach advocated is four gland parathyroidectomy without reimplantation. The aim of this study was to review the medium term results of this procedure. METHODS: Fifty four consecutive patients with end stage chronic renal failure and secondary hyperparathyroidism who had a four gland parathyroidectomy without reimplantation were studied. The procedure was performed by a single surgeon with a median (range) follow up of 29 (0-70) months. RESULTS: Most patients (76%) developed postoperative hypocalcaemia but this was easily treated and doses of long term drugs necessary to prevent this were low. Pre-operative bone symptoms, hypercalcaemia, hyperphosphataemia, and an increased alkaline phosphatase were improved or resolved in most patients. Thirteen (24%) patients had an undetectable postoperative parathyroid hormone (PTH), (6 of 12 (50%) with a functioning renal transplant and 7 of 42 (17%) who required dialysis, p = 0.02). Median (range) postoperative PTH values in these groups were 0.1 (0.1-31) compared with 1.0 (0.1-24) pmol/l (p = 0.085) respectively. The remaining 41 of 54 (76%) patients had residual PTH secretion and postoperative hyperparathyroidism was identified in eight (15%) patients with only two requiring neck re-exploration. CONCLUSION: Four gland parathyroidectomy without reimplantation produced good medium term biochemical and clinical results. Most patients had minor residual PTH secretion that may contribute to this and mitigate concerns regarding adynamic bone disease. Endogenous PTH secretion is only completely lost in a few patients but occurs more often in those with a functioning renal transplant. Bone densitometry is required to investigate the long term impact of this procedure. PMID- 15811892 TI - Peritoneovenous shunting is an effective treatment for intractable ascites. AB - AIM AND METHODS: A retrospective review was carried out of children undergoing peritoneovenous shunting for intractable ascites. RESULTS: 11 children, aged 3 months to 12 years (median 31 months) underwent peritoneovenous shunting over the past 17 years. The duration of ascites ranged from one month to 2.5 years (median two months). The primary pathology consisted of previous surgery in eight (three neuroblastoma, one renal carcinoma, one hepatoblastoma, one adrenal teratoma, one renal artery stenosis, and one diaphragmatic hernia), and cytomegalovirus hepatitis, lymphatic hypoplasia, and lymphohistiocytosis in one patient each. All patients had failed to respond to previous treatment including peritoneal drainage in six patients, diuretics in five, and parenteral nutrition in five. There were no intraoperative problems. Postoperative complications included pulmonary oedema in three patients, shunt occlusion in three, infection in two, and wound leakage in one. Ascites resolved after shunting in 10 patients. Five shunts were removed one to three years after insertion without recurrence of ascites. Three others are free of ascites with shunts in place for less than one year postoperatively. Three children died from their underlying disease: two after resolution of ascites (neuroblastoma) and one in whom the ascites failed to resolve (lymphohisticytosis). CONCLUSIONS: Peritoneovenous shunting is an effective treatment for symptomatic intractable ascites in children (10 of 11 successful cases in this series). Elective removal of the shunt after one year is recommended. PMID- 15811893 TI - Correlations of work, leisure, and sports physical activities and health status with socioeconomic factors: a national study in Israel. AB - OBJECTIVE: To evaluate levels of physical activities at work, leisure, and sports and to correlate them with socioeconomic and health factors. METHODS: Validated questionnaire administered to a random sample of 406 adults. Items covered demographic data, health status, smoking, and duration, frequency, intensity of physical activities. Indices of physical activity at work, leisure, and sports were analysed. RESULTS: Adults (both sexes) with poor self perceived health status and less than 13 years of education, regardless of their body mass index, perform no or few physical activities during their leisure time. CONCLUSIONS: The correlations of physical activity with socioeconomic and health factors differ significantly for work, leisure, and sports. Physicians should differentiate physical activities by type and intensity during anamneses. PMID- 15811894 TI - Retinal vein thrombosis associated with a herbal phytoestrogen preparation in a susceptible patient. AB - A 55 year old woman, known to have systemic lupus erythematosus and protein S deficiency, presented with sudden onset of visual loss in the left eye. Three days previously, she took a herbal phytoestrogen preparation for relief of her menopausal symptoms. Ophthalmological assessment showed a left inferior hemiretinal vein thrombosis. The temporal relation between the onset of visual symptoms and ingestion of this herbal preparation suggested that in this patient with a prior non-manifest prothrombotic tendency, there may be an association between the development of retinal vein thrombosis and herbal phytoestrogen use. PMID- 15811895 TI - Recurrent acute heart failure caused by sliding hiatus hernia. AB - The case is reported of a 75 year old woman who presented with recurrent nocturnal episodes of acute pulmonary oedema. The cause was uncertain as she had normal cardiothoracic ratio on chest radiography and normal left ventricular systolic and diastolic function by transthoracic echocardiogram. Another transthoracic echocardiogram was repeated when she was recumbent for an hour and had a full stomach. It showed a striking finding of severe left atrial compression by an external structure. Computed tomography of the thorax showed an intrathoracic mass behind the left atrium causing external compression of the left atrium suggestive of a sliding hiatus hernia. Cardiac catheterisation confirmed the diagnosis by showing a pronounced rise of pulmonary capillary wedge pressure in the recumbent position compared with the sitting up position. PMID- 15811896 TI - Acute liver failure: a message found under the skin. AB - Acute liver failure is a rare syndrome with rapid progression and high mortality. It is characterised by the onset of coma and coagulopathy usually within six weeks but can occur up to six months after the onset of illness. Viral hepatitis, idiosyncratic drug induced liver injury, and acetaminophen ingestion are common causes. This report describes the case of a 35 year old man who presented with acute liver failure shortly after binge drinking. Repeated history taking disclosed a gluteal disulfiram implant that the patient had received to treat his alcohol dependence. The patient recovered with maximum supportive care after surgical removal but without liver transplantation. This case illustrates that only meticulous history taking will disclose the sometimes bewildering causes of acute liver failure. PMID- 15811897 TI - Hyperpigmented rugosity of skin associated with adenocarcinoma of the caecum. PMID- 15811898 TI - High-dose insulin therapy for calcium-channel blocker overdose. AB - OBJECTIVE: To evaluate the evidence for using high-dose insulin therapy with supplemental dextrose and potassium in calcium-channel blocker (CCB) overdose. DATA SOURCES: Evidence of efficacy for high-dose insulin therapy with supplemental dextrose and potassium was sought by performing a search of MEDLINE and Toxline between 1966 and July 2004 using combinations of the terms calcium channel blocker, overdose, poisoning, antidote, and insulin. Abstracts from the North American Congress of Clinical Toxicology for the years 1996-2003 were also reviewed. STUDY SELECTION AND DATA EXTRACTION: Identified articles, including animal studies, case reports, and case series, were evaluated for this review. No clinical trials were available. DATA SYNTHESIS: Animal models of CCB overdose demonstrate that high-dose insulin with supplemental dextrose and potassium was a more effective therapy than calcium, glucagon, or catecholamines. High-dose insulin appears to enhance cardiac carbohydrate metabolism and has direct inotropic effects. Published clinical experience is limited to 13 case reports where insulin was used after other therapies were failing; 12 of these patients survived. High-dose insulin therapy was beneficial for CCB-induced hypotension, hyperglycemia, and metabolic acidosis. Bradycardia and heart block resolved in some patients, but persisted in others. CONCLUSIONS: Based on animal data and limited human experience, as well as the inadequacies of available alternatives for patients with significant poisoning, high-dose insulin therapy warrants further study and judicious use in patients with life-threatening CCB poisoning. PMID- 15811899 TI - Initializing concordance in frail elderly patients via a medicines organizer. AB - BACKGROUND: Concordance, which involves prescribing with rather than for patients, should result in less drug wastage (non-use), but is proving hard to put into practice. One possible way of easing elderly people and prescribers toward concordance is to use a medicines organizer (MO). OBJECTIVE: To document medication wastage, using a pharmacy-prepared reusable MO, and explore, using a qualitative approach, use of this information on communication of individuals' drug regimens. METHODS: Sixty-two sheltered housing residents, aged > or =60 years, participated in an exploratory controlled, matched study. The intervention group received medication in the MO, and the control group continued with standard packaging. Community pharmacists recorded details of wasted drugs returned during the 3-month study and, for the intervention group, 6, 9, and 12 months after the study. Medicines management data were collected from participants. Qualitative interviews were conducted with a sample of general practitioners (GPs) and community pharmacists to explore views on the role of the MO. RESULTS: Intervention group wastage was reduced from 18.1% (prestudy) to 1% at 12 months. No data on wastage were collected for the control group after the prestudy assessment. Sixty-one percent more prescription changes, including significant decreases in the number of prescribed drugs and dosages, were reported for the intervention group. GPs and pharmacists reported improvements in communication concerning medication-related dialog. CONCLUSIONS: This small exploratory study has shown that a pharmacy-prepared reusable MO provided visual, objective insights into medication wastage. This resulted in improved communication of drug needs and reduced wastage-the foundation for concordance. PMID- 15811900 TI - Impact of stress ulcer prophylaxis algorithm study. AB - BACKGROUND: In the intensive care unit at Royal Victoria Hospital, we noted that drugs prescribed for stress ulcer prophylaxis were not always indicated or optimal. Accordingly, we implemented an algorithm for stress ulcer prophylaxis to guide the medical team in their decisions. The agents selected for the algorithm were intravenous famotidine and omeprazole suspension or tablets, depending on the available administration route. OBJECTIVE: To evaluate the impact of a treatment algorithm on the appropriateness of prescriptions for stress ulcer prophylaxis. METHODS: A quasi-experimental-type evaluative study was conducted based on a pre-/post-intervention design without a concurrent control group. A total of 555 complete admissions met the selection criteria; 303 patients formed the pre-intervention group, and 252 made up the post-intervention group (exposed to the treatment algorithm). RESULTS: After implementation of the algorithm, the proportion of inappropriate prophylaxis was decreased (95.7% vs 88.2%; p = 0.033). The number of days of inappropriate prophylaxis was also reduced significantly (p = 0.013), as was the cost per patient (p = 0.003) for all admissions. However, no difference was observed when the subgroup of patients who received prophylaxis alone was studied (p = 0.098 and p = 0.918). The presence of bleeding was similar in both groups. CONCLUSIONS: Introduction by pharmacists of a treatment algorithm for stress ulcer prophylaxis in intensive care units allows a reduction of inappropriate prescriptions and thus a reduction in the cost of drugs. The use of omeprazole suspension seems to be an alternative to intravenous histamine2-inhibitors; however, a large-scale study is necessary to confirm the efficacy and safety of proton-pump inhibitors administered by an enteral tube. PMID- 15811902 TI - Comment: insulin strategies for type 2 diabetes mellitus. PMID- 15811903 TI - Clinical experience with spironolactone in pediatrics. AB - BACKGROUND: In 2003, the Food and Drug Administration placed spironolactone on its list of drugs needing pediatric studies. OBJECTIVE: To describe the use of spironolactone in a large group of children and evaluate its safety, focusing on its effects on potassium. METHODS: A prospective observational study was conducted. Patient demographic information was collected, as well as dosing regimens, use of other medications, and potassium concentrations. Patients were grouped by diagnosis. Comparisons were made with unpaired t-tests. RESULTS: One hundred consecutive patients were evaluated. The average age was 20.8 months and weight was 9.5 kg. Sixty-two patients had heart disease (HD), 29 had chronic lung disease (CLD), and 9 had other conditions. The initial dose was 1.8 +/- 0.7 mg/kg/day. Patients with CLD received a higher dose than those with HD (2 +/- 0.8 vs 1.7 +/- 0.5 mg/kg/day; p = 0.04). Sixty-six patients received furosemide and 37 received thiazides (12 received both). The average potassium concentration after initiation was 4.3 +/- 0.8 mEq/L, with higher values in patients with CLD versus HD (4.7 +/- 0.7 vs 4.2 +/- 0.7 mEq/L; p = 0.007). Twenty-six patients required potassium supplementation, including 16 with CLD and 8 with HD; no other adverse effects were noted. Average length of treatment was 16 days, with a length of stay of 38 days. Of the 92 patients surviving to discharge, 66 continued on spironolactone. CONCLUSIONS: This sample demonstrates that spironolactone is a common component of diuretic regimens in pediatric patients. The only adverse effects were alterations in potassium. While hyperkalemia was more common initially, hypokalemia was more frequent with long-term use. Potassium concentrations should be carefully monitored, particularly in children receiving multiple diuretics. Additional research is needed to define the pharmacokinetics and optimal dosing interval of spironolactone, as well as determine its long-term effects on potassium. PMID- 15811904 TI - What is new with the beta2-agonists: issues in the management of asthma. AB - OBJECTIVE: To review the more recent literature addressing the issue of whether beta2-agonists can worsen asthma and/or increase the risk of severe exacerbations and death from asthma. DATA SOURCES: PubMed was searched (2001-December 2004), along with the Food and Drug Administration and Cochrane Library Web sites. In addition, the bibliographies of recent reviews of the subject were assessed. STUDY SELECTION AND DATA EXTRACTION: Randomized clinical trials, retrospective and prospective cohort studies, and meta-analyses published in the past 3 years were reviewed. Studies assessing the potential for beta2-agonists to worsen outcomes in asthma as well as long-term studies assessing asthma outcomes that included an arm with regular administration of short- or long-acting inhaled beta2-agonists (LABAs) were selected. Worsening asthma was defined as a decline in lung function, an increase in bronchial hyperresponsiveness, exacerbations, or death. Studies older than 3 years selected from the bibliographies of the primary articles that addressed background perspective were also included where appropriate. DATA SYNTHESIS: The studies fell into 3 primary categories with some overlap: those assessing toxicity of the S-enantiomer of albuterol, those evaluating the risk of specific genotypes regarding worsening asthma, and those assessing asthma outcomes with LABA therapy. CONCLUSIONS: The current data on the use of beta2-agonists continue to support the national and international guidelines for the treatment of asthma. That is, short-acting inhaled beta2 agonists should only be used as needed for symptoms and prevention of exercise induced bronchospasm, and LABAs should only be used regularly as adjunctive therapy with inhaled corticosteroids in patients whose asthma is not controlled with low to medium doses of the inhaled corticosteroid. PMID- 15811905 TI - Dyspnea during thalidomide treatment for advanced ovarian cancer. AB - OBJECTIVE: To detail the dyspnea encountered in women receiving thalidomide as therapy for advanced ovarian cancer. CASE SUMMARIES: Eight of 18 (44%) patients with recurrent ovarian cancer developed dyspnea while receiving thalidomide 200 mg daily as part of a prospective Phase II study. Dyspnea was evaluated with pulse oximetry, chest X-ray and, if indicated, spiral computed tomography scan. Four patients had abnormal chest X-ray findings (1 pleural effusion, 1 pneumonia, 2 mild congestive heart failure), and one of these patients also had a pulmonary embolus. The other 4 patients had no objective test findings to explain their dyspnea. Five patients had resolution of symptoms when thalidomide was discontinued and, when the drug was resumed at a 50% dose reduction, experienced no further shortness of breath. DISCUSSION: While dyspnea in association with thalidomide has not previously been reported as a common adverse event, it was a frequent complaint of patients receiving this drug as part of a Phase II study. Comorbid conditions causing dyspnea were evaluated since they are common in this patient population; however, half of our patients had no objective evidence of such conditions. The Naranjo probability scale indicated a probable relationship between dyspnea and thalidomide therapy in the patients with no objective evidence of comorbidity. We advocate discontinuation of thalidomide until symptoms have resolved, at which time reintroduction of thalidomide at a reduced dose may be considered. CONCLUSIONS: Patients receiving thalidomide may develop dyspnea as an adverse effect of the drug. In selected patients, thalidomide may be safely reintroduced once symptoms resolve. PMID- 15811906 TI - Angioedema associated with aspirin and rofecoxib. AB - OBJECTIVE: To report the probable association of angioedema with aspirin therapy and the selective cyclooxygenase-2 (COX-2) inhibitor rofecoxib. CASE SUMMARY: A 44-year-old white woman, previously tolerant to aspirin and other nonsteroidal antiinflammatory drugs (NSAIDs), developed angioedema of the lips after ingesting two 325-mg aspirin tablets during one day. The reaction occurred 3 hours after taking the second aspirin and resolved within 3 hours. Two weeks later, the patient took a 25-mg rofecoxib tablet for a sore throat, and she developed angioedema 5(1/2) hours later. Although the woman took 50 mg of diphenhydramine, the swelling did not subside. She repeated the diphenhydramine dose in the evening and, by noon the next day, 26(1/2) hours after the angioedema began, it was resolved. The patient's internist prescribed an epinephrine auto-injector and advised her to consult an allergist. With skin testing and oral rechallenge with aspirin, but not rofecoxib, the allergist determined the cause of the reactions to be aspirin-induced angioedema and selective COX-2 inhibitor intolerance. The Naranjo probability scale indicated that aspirin was a highly probable cause and rofecoxib was a probable cause of this patient's angioedema. DISCUSSION: Aspirin induced angioedema and NSAID intolerance have been well documented. There are reports of both tolerance and intolerance to selective COX-2 inhibitors in patients with documented allergy-like reactions to aspirin and NSAIDs. CONCLUSIONS: Patients with aspirin and NSAID intolerance may develop intolerance to COX-2 inhibitors, especially with repeated exposure. PMID- 15811907 TI - Inhaled corticosteroids and rate of exacerbations in patients with COPD. PMID- 15811908 TI - Pheochromocytoma unmasked by amisulpride and tiapride. AB - OBJECTIVE: To describe the unmasking of pheochromocytoma in a patient treated with amisulpride and tiapride. CASE SUMMARY: A 42-year-old white man developed acute hypertension with severe headache and vomiting 2 hours after the first doses of amisulpride 100 mg and tiapride 100 mg. Both drugs were immediately discontinued, and the patient recovered after subsequent nicardipine and verapamil treatment. Abdominal ultrasound showed an adrenal mass, and postoperative histologic examination confirmed the diagnosis of pheochromocytoma. DISCUSSION: Drug-induced symptoms of pheochromocytoma are often associated with the use of substituted benzamide drugs, but the underlying mechanism is unknown. In our case, use of the Naranjo probability scale indicated a possible relationship between the hypertensive crisis and amisulpride and tiapride therapy. CONCLUSIONS: As of March 24, 2005, this is the first reported case of amisulpride- and tiapride-induced hypertensive crisis in a patient with pheochromocytoma. Physicians and other healthcare professionals should be aware of this potential adverse effect of tiapride and amisulpride. PMID- 15811909 TI - Intrathecal colistin and sterilization of resistant Pseudomonas aeruginosa shunt infection. AB - OBJECTIVE: To report 2 cases of multidrug-resistant (MDR) Pseudomonas aeruginosa meningitis and ventriculo-peritoneal shunt (VPS) infection successfully sterilized with intrathecal colistin 10 mg/day after development of nephrotoxicity associated with intravenous administration. CASE SUMMARIES: Case 1. A 69-year-old African American woman with a history of subarachnoid hemorrhage and hydrocephalus requiring VPS placement was admitted with VPS infection and meningitis. Cerebrospinal fluid (CSF) cultures revealed MDR P. aeruginosa susceptible only to colistin. Intravenous colistin was initiated but rapidly discontinued due to development of renal dysfunction. Intravenous colistin was the probable cause of the adverse effect. Intrathecal colistin was initiated via an externalized VPS, with subsequent improvement in white blood cell counts in the CSF. Follow-up CSF cultures remained sterile and renal function returned to baseline. Case 2. A 69-year-old white woman with a history of subarachnoid hemorrhage, hydrocephalus, and VPS was transferred from an extended-care facility for management of a VPS infection. CSF cultures revealed MDR P. aeruginosa susceptible only to colistin. Intravenous colistin was initiated but subsequently discontinued due to worsening renal function that, as with the first case, probably correlated with colistin administration and persisted despite dose adjustment. Therapy was changed to intrathecal administration, with subsequent normalization of her CSF white blood cell counts and sterilization of cultures. DISCUSSION: The limited availability of antibiotics for treatment of highly resistant or MDR gram-negative organisms has prompted clinicians to reconsider the use of older drugs. Prior reports have suggested that intravenous colistin is a potential alternative for treating highly resistant gram-negative central nervous system infections, specifically Acinetobacter, but its use is limited by nephrotoxicity. Our experience suggests that intrathecal colistin is a potentially curative intervention for the treatment of severe MDR P. aeruginosa meningitis and VPS infections in patients in whom intravenous colistin is not an option. CONCLUSIONS: Intrathecal use of colistin is a potentially safe, effective, and viable treatment option for MDR P. aeruginosa central nervous system infections when intravenous administration is not feasible. PMID- 15811910 TI - High-dose clopidogrel loading in percutaneous coronary intervention. AB - OBJECTIVE: To review the use of a 600-mg clopidogrel loading dose in patients undergoing percutaneous coronary intervention (PCI). DATA SOURCES: Human clinical trials and platelet studies available through PubMed (1966-March 2005), bibliographies of pertinent articles, and citations supplied by the drug manufacturer were accessed. DATA SYNTHESIS: The administration of a 600-mg loading dose of clopidogrel can decrease the time required for maximum platelet inhibition to 2 hours compared with > or =6 hours achieved with 300 mg. This higher loading dose has been investigated in multiple platelet studies and one observational report. Several randomized controlled trials have used a 600-mg loading dose; however, these studies were not designed to evaluate the efficacy and safety of this loading regimen. To date, only one randomized trial has compared the 600-mg loading dose with a 300-mg loading dose. CONCLUSIONS: When compared with a conventional loading regimen of 300 mg in lower-risk patients, pretreatment with clopidogrel 600 mg was shown to be more effective in reducing periprocedural events and demonstrated similar safety. Studies are needed to clarify the use of a 600-mg loading dose in higher-risk patients, with concomitant glycoprotein IIb/IIIa receptor antagonism, or when administration is delayed until immediately before or after PCI. PMID- 15811911 TI - Meeting report: the future and limits of systems biology. AB - Systems biology has enjoyed explosive growth in both the number of people participating in this area of research and the number of publications on the topic. And yet, the paradigms that underlie the field have not seen a similar expansiveness. Instead, most of these paradigms have been carried over from other fields like engineering, physics, and mathematics. As a result, a small set of concepts dominate the field. The traditional biologist is seen by many as outmoded and tolerated only as a source of data. In this view, the biologist's ideas may even be considered conceptually and theoretically irrelevant. In this Perspective, we take a critical look at some of the paradigms of systems biology and question whether the biologist's ideas, methods, and theories have really become outmoded. We see the future of systems biology as a tight coupling of in vivo and in vitro methods for bioengineering with in silico multicellular modeling and simulation. PMID- 15811912 TI - The N- and C-terminal RNA recognition motifs of splicing factor Prp24 have distinct functions in U6 RNA binding. AB - Prp24 is an essential yeast U6 snRNP protein with four RNA recognition motifs (RRMs) that facilitates the association of U4 and U6 snRNPs during spliceosome assembly. Genetic interactions led to the proposal that RRMs 2 and 3 of Prp24 bind U6 RNA, while RRMs 1 and 4 bind U4 RNA. However, the function of each RRM has yet to be established through biochemical means. We compared the binding of recombinant full-length Prp24 and truncated forms lacking RRM 1 or RRM 4 with U6 RNA. Contrary to expectations, we found that the N-terminal segment containing RRM 1 is important for high-affinity binding to U6 RNA and for discrimination between wild-type U6 RNA and U6 with point mutations in the 3' intramolecular stem-loop. In contrast, deletion of RRM 4 and the C terminus did not significantly alter the affinity for U6 RNA, but resulted in the formation of higher order Prp24.U6 complexes. Truncation and internal deletion of U6 RNA mapped three Prp24-binding sites, with the central site providing most of the affinity for Prp24. A newly identified temperature-sensitive lethal point mutation in RRM 1 is exacerbated by mutations in the U6 RNA telestem, as is a mutation in RRM 2, but not one in RRM 3. We propose that RRMs 1 and 2 of yeast Prp24 bind the same central site in U6 RNA that is bound by the two RRMs of human Prp24, and that RRMs 3 and 4 bind lower affinity flanking sites, thereby restricting the stoichiometry of Prp24 binding. PMID- 15811913 TI - tRNA m7G methyltransferase Trm8p/Trm82p: evidence linking activity to a growth phenotype and implicating Trm82p in maintaining levels of active Trm8p. AB - We show that Saccharomyces cerevisiae strains lacking Trm8p/Trm82p tRNA m7G methyltransferase are temperature-sensitive in synthetic media containing glycerol. Bacterial TRM8 orthologs complement the growth defect of trm8-Delta, trm82-Delta, and trm8-Delta trm82-Delta double mutants, suggesting that bacteria employ a single subunit for Trm8p/Trm82p function. The growth phenotype of trm8 mutants correlates with lack of tRNA m7G methyltransferase activity in vitro and in vivo, based on analysis of 10 mutant alleles of trm8 and bacterial orthologs, and suggests that m7G modification is the cellular function important for growth. Initial examination of the roles of the yeast subunits shows that Trm8p has most of the functions required to effect m7G modification, and that a major role of Trm82p is to maintain cellular levels of Trm8p. Trm8p efficiently cross-links to pre-tRNAPhe in vitro in the presence or absence of Trm82p, in addition to its known residual tRNA m7G modification activity and its SAM-binding domain. Surprisingly, the levels of Trm8p, but not its mRNA, are severely reduced in a trm82-Delta strain. Although Trm8p can be produced in the absence of Trm82p by deliberate overproduction, the resulting protein is inactive, suggesting that a second role of Trm82p is to stabilize Trm8p in an active conformation. PMID- 15811914 TI - Molecular dynamics simulations and free energy calculations of base flipping in dsRNA. AB - The family of adenosine deaminases acting on RNA (ADARs) targets adenosines in RNA that is mainly double stranded. Some substrates are promiscuously deaminated whereas others, such as the mammalian glutamate receptor B (gluR-B) pre-mRNA, are more selectively deaminated. Many DNA/RNA-base modification enzymes use a base flipping mechanism to be able to reach their target base and it is believed that ADARs function in a similar way. In this study we used molecular dynamics (MD) simulations to describe two sites on the gluR-B pre-mRNA, the selectively targeted R/G site and the nontargeted 46 site, in an attempt to explain the substrate specificity. We used regular MD and also a forced base flipping method with umbrella sampling to calculate the free energy of base opening. Spontaneous opening of the mismatched adenosine was observed for the R/G site but not for the 46 site. PMID- 15811915 TI - Structural implications of novel diversity in eucaryal RNase P RNA. AB - Previous eucaryotic RNase P RNA secondary structural models have been based on limited diversity, representing only two of the approximately 30 phylogenetic kingdoms of the domain Eucarya. To elucidate a more generally applicable structure, we used biochemical, bioinformatic, and molecular approaches to obtain RNase P RNA sequences from diverse organisms including representatives of six additional kingdoms of eucaryotes. Novel sequences were from acanthamoeba (Acathamoeba castellanii, Balamuthia mandrillaris, Filamoeba nolandi), animals (Caenorhabditis elegans, Drosophila melanogaster), alveolates (Theileria annulata, Babesia bovis), conosids (Dictyostelium discoideum, Physarum polycephalum), trichomonads (Trichomonas vaginalis), microsporidia (Encephalitozoon cuniculi), and diplomonads (Giardia intestinalis). An improved alignment of eucaryal RNase P RNA sequences was assembled and used for statistical and comparative structural analysis. The analysis identifies a conserved core structure of eucaryal RNase P RNA that has been maintained throughout evolution and indicates that covariation in size occurs between some structural elements of the RNA. Eucaryal RNase P RNA contains regions of highly variable length and structure reminiscent of expansion segments found in rRNA. The eucaryal RNA has been remodeled through evolution as a simplified version of the structure found in bacterial and archaeal RNase P RNAs. PMID- 15811916 TI - In vivo aggregation properties of the nuclear poly(A)-binding protein PABPN1. AB - A broad range of degenerative diseases is associated with intracellular inclusions formed by toxic, aggregation-prone mutant proteins. Intranuclear inclusions constitute a pathological hallmark of oculopharyngeal muscular dystrophy (OPMD), a dominantly inherited disease caused by (GCG) repeat expansions in the gene that encodes for nuclear poly(A) binding protein (PABPN1). The mutation results in an extended polyalanine stretch that has been proposed to induce protein aggregation and formation of intranuclear inclusions. Here we show that normal PABPN1 is inherently aggregation-prone when exogenously expressed in either HeLa or myogenic C2 cells. Similar deposits of insoluble PABPN1 are formed by variant forms of the protein containing either a polyalanine expansion or a complete deletion of the polyalanine tract, indicating that the mutation responsible for OPMD is not essential for formation of PABPN1 inclusions. In contrast, interfering with any of the protein domains required for stimulation of poly(A) polymerase prevents the formation of inclusions. Most surprisingly, photobleaching experiments reveal that both normal and expanded PABPN1 molecules are not irreversibly sequestered into aggregates, but rather move rapidly in and out of the inclusions. These findings have important implications for the interpretation of OPMD model systems based on exogenous expression of PABPN1. PMID- 15811917 TI - Nonbridging phosphate oxygens in 16S rRNA important for 30S subunit assembly and association with the 50S ribosomal subunit. AB - Ribosomes are composed of RNA and protein molecules that associate together to form a supramolecular machine responsible for protein biosynthesis. Detailed information about the structure of the ribosome has come from the recent X-ray crystal structures of the ribosome and the ribosomal subunits. However, the molecular interactions between the rRNAs and the r-proteins that occur during the intermediate steps of ribosome assembly are poorly understood. Here we describe a modification-interference approach to identify nonbridging phosphate oxygens within 16S rRNA that are important for the in vitro assembly of the Escherichia coli 30S small ribosomal subunit and for its association with the 50S large ribosomal subunit. The 30S small subunit was reconstituted from phosphorothioate substituted 16S rRNA and small subunit proteins. Active 30S subunits were selected by their ability to bind to the 50S large subunit and form 70S ribosomes. Analysis of the selected population shows that phosphate oxygens at specific positions in the 16S rRNA are important for either subunit assembly or for binding to the 50S subunit. The X-ray crystallographic structures of the 30S subunit suggest that some of these phosphate oxygens participate in r-protein binding, coordination of metal ions, or for the formation of intersubunit bridges in the mature 30S subunit. Interestingly, however, several of the phosphate oxygens identified in this study do not participate in any interaction in the mature 30S subunit, suggesting that they play a role in the early steps of the 30S subunit assembly. PMID- 15811918 TI - UTP-dependent turnover of Trypanosoma brucei mitochondrial mRNA requires UTP polymerization and involves the RET1 TUTase. AB - Trypanosoma brucei mitochondria possess a unique RNA decay pathway in which rapid degradation of polyadenylated mRNAs is dependent on the addition of UTP, as measured by in organello pulse chase assays. To determine the mechanism by which UTP stimulates the degradation of polyadenylated RNAs, we performed in organello pulse chase assays under different conditions. Treatment of mitochondria with proteinase K revealed that UTP does not act through a receptor on the surface of the mitochondria. To determine if the UTP-stimulated RNA decay pathway is triggered by the mitochondrial energy state or ATP:UTP ratio, increasing ATP was added to a constant amount of UTP during the chase period of the assay. Results indicate that rapid turnover is responsive to UTP and not the ATP:UTP ratio. Experiments using UTP analogs demonstrate that UTP polymerization into RNAs is necessary for UTP-dependent degradation. Furthermore, experiments performed with RNAi cells indicate that the RET1 terminal uridylyl transferase (TUTase) is required for UTP-dependent decay of polyadenylated RNAs. Overall, these results show that degradation of polyadenylated RNAs in T. brucei mitochondria can occur through a unique mechanism that requires the polymerization of UTP into RNAs, presumably by the RET1 TUTase. PMID- 15811919 TI - Transcription and RNA-processing in fission yeast mitochondria. AB - We systematically examined transcription and RNA-processing in mitochondria of the petite-negative fission yeast Schizosaccharomyces pombe. Two presumptive transcription initiation sites at opposite positions on the circular-mapping mtDNA were confirmed by in vitro capping of primary transcripts with guanylyl transferase. The major promoter (Pma) is located adjacent to the 5'-end of the rnl gene, and a second, minor promoter (Pmi) upstream from cox3. The primary 5' termini of the mature rnl and cox3 transcripts remain unmodified. A third predicted accessory transcription initiation site is within the group IIA1 intron of the cob gene (cobI1). The consensus promoter motif of S. pombe closely resembles the nonanucleotide promoter motifs of various yeast mtDNAs. We further characterized all mRNAs and the two ribosomal RNAs by Northern hybridization, and precisely mapped their 5'- and 3'-ends. The mRNAs have leader sequences with a length of 38 up to 220 nt and, in most instances, are created by removal of tRNAs from large precursor RNAs. Like cox2 and rnl, cox1 and cox3 are not separated by tRNA genes; instead, transcription initiation from the promoters upstream from rnl and cox3 compensates for the lack of tRNA-mediated 5'-processing. The 3' termini of mRNAs and of SSU rRNA are processed at distinct, C-rich motifs that are located at a variable distance (1-15 nt) downstream from mRNA and SSU-rRNA coding regions. The accuracy of RNA-processing at these sites is sequence dependent. Similar 3'-RNA-processing motifs are present in species of the genus Schizosaccharomyces, but not in budding yeasts that have functionally analogous A+T-rich dodecamer processing signals. PMID- 15811920 TI - Transfer-messenger RNA unfolds as it transits the ribosome. AB - In bacteria, translation of mRNAs lacking stop codons produces truncated polypeptides and traps ribosomes in unproductive complexes. Potentially harmful truncated proteins are tagged with short peptides encoded by the mRNA-like domain of tmRNA and targeted for digestion by housekeeping proteases. We show that altered Escherichia coli transfer-messenger RNAs (tmRNAs) produce in vivo fusion proteins with peptide tags that extend far beyond the conventional termination signal of the wild-type tmRNA. Regions of tmRNA capable of serving as templates for protein synthesis include helix 5, as well as pseudoknots 2, 3, and 4. The removal of all six in-frame stop codons negatively affects tmRNA processing, thereby preventing translation of the 3' portion of the tRNA-like domain. These findings provide evidence that trans-translation can be accompanied by the unfolding of significant portions of the tmRNA molecule. Many of these conformational changes are likely to be required during trans-translation to maintain the ribosomal subunits in close proximity to the tmRNA for monitoring its transit. PMID- 15811921 TI - The contributions of dsRNA structure to Dicer specificity and efficiency. AB - Dicer processes long double-stranded RNA (dsRNA) and pre-microRNAs to generate the functional intermediates (short interfering RNAs and microRNAs) of the RNA interference pathway. Here we identify features of RNA structure that affect Dicer specificity and efficiency. The data presented show that various attributes of the 3' end structure, including overhang length and sequence composition, play a primary role in determining the position of Dicer cleavage in both dsRNA and unimolecular, short hairpin RNA (shRNA). We also demonstrate that siRNA end structure affects overall silencing functionality. Awareness of these new features of Dicer cleavage specificity as it is related to siRNA functionality provides a more detailed understanding of the RNAi mechanism and can shape the development of hairpins with enhanced functionality. PMID- 15811922 TI - 6S RNA is a widespread regulator of eubacterial RNA polymerase that resembles an open promoter. AB - 6S RNA is an abundant noncoding RNA in Escherichia coli that binds to sigma70 RNA polymerase holoenzyme to globally regulate gene expression in response to the shift from exponential growth to stationary phase. We have computationally identified >100 new 6S RNA homologs in diverse eubacterial lineages. Two abundant Bacillus subtilis RNAs of unknown function (BsrA and BsrB) and cyanobacterial 6Sa RNAs are now recognized as 6S homologs. Structural probing of E. coli 6S RNA and a B. subtilis homolog supports a common secondary structure derived from comparative sequence analysis. The conserved features of 6S RNA suggest that it binds RNA polymerase by mimicking the structure of DNA template in an open promoter complex. Interestingly, the two B. subtilis 6S RNAs are discoordinately expressed during growth, and many proteobacterial 6S RNAs could be cotranscribed with downstream homologs of the E. coli ygfA gene encoding a putative methenyltetrahydrofolate synthetase. The prevalence and robust expression of 6S RNAs emphasize their critical role in bacterial adaptation. PMID- 15811923 TI - Co-evolution of tRNA 3' trailer sequences with 3' processing enzymes in bacteria. AB - Maturation of the tRNA 3' terminus is a complicated process in bacteria. Usually, it is initiated by an endonucleolytic cleavage carried out by RNase E and Z in different bacteria. In Escherichia coli, RNase E cleaves AU-rich sequences downstream of tRNA, producing processing intermediates with a few extra residues at the 3' end; these are then removed by exoribonuclease trimming to generate the mature 3' end. Here we show that essentially all E. coli tRNA precursors contain a potential RNase E cleavage site, the AU-rich sequence element (AUE), in the 3' trailer. This suggests that RNase E cleavage and exonucleolytic trimming is a general pathway for tRNA maturation in this organism. Remarkably, the AUE immediately downstream of each tRNA is selectively conserved in bacteria having RNase E and tRNA-specific exoribonucleases, suggesting that this pathway for tRNA processing is also commonly used in these bacteria. Two types of RNase E-like proteins are identified in actinobacteria and the alpha-subdivision of proteobacteria. The tRNA 3' proximal AUE is conserved in bacteria with only one type of E-like protein. Selective conservation of the AUE is usually not observed in bacteria without RNase E. These results demonstrate a novel example of co evolution of RNA sequences with processing activities. PMID- 15811924 TI - 17alpha-hydroxylase/17,20-lyase deficiency caused by a novel homozygous mutation (Y27Stop) in the cytochrome CYP17 gene. AB - CONTEXT: 17alpha-Hydroxylase/17,20-lyase deficiency, a rare autosomal recessive form of congenital adrenal hyperplasia, is caused by mutations in the cytochrome P450c17 (CYP17) gene. We report on a case of complete 17alpha-hydroxylase/17,20 lyase deficiency due to a novel homozygous mutation of CYP17. DESIGN: A 20-yr-old female Turkish patient (46,XX) presented with primary amenorrhea, sexual infantilism, and easy fatigability. RESULTS: The patient's steroid metabolism showed increased levels of mineralocorticoid precursors and low or undetectable plasma concentrations of 17alpha-hydroxycorticoids, androgens, and estrogens before and after ACTH stimulation. The gas chromatography-mass spectrometry urinary steroid profile was dominated by metabolites of corticosterone and its precursors, while cortisol and C(19)-steroid metabolites were lacking. ACTH, FSH, and LH levels were elevated. These hormonal findings were consistent with a combined and total 17alpha-hydroxylase/17,20-lyase deficiency. A therapy with hydrocortisone and a cyclic estrogen/gestagen substitution was initiated. CONCLUSION: The CYP17 gene analysis revealed homozygosity of the mutation Y27Stop (TAC-->TAA) in exon 1, a mutation that has not been previously described. This novel mutation leads to a stop codon causing a total loss of 17alpha hydroxlyase/17,20-lyase activity, as reflected biochemically by the detected concentrations of the steroid metabolites. PMID- 15811925 TI - Spontaneous normalization of thyrotropin concentrations in patients with subclinical hypothyroidism. AB - CONTEXT: Patients with subclinical hypothyroidism may revert to normal TSH values. OBJECTIVE: The objective of this study was to examine the time course of the normalization of TSH levels in subclinical hypothyroidism. DESIGN: This was a prospective, observational study with no intervention, with a duration of follow up of 12-72 months. SETTING: Outpatients visited an endocrinology clinic of a general hospital. PATIENTS: Forty patients (32 women, mean age 62.8 +/- 8.2 yr) with spontaneous subclinical hypothyroidism (TSH > 5 mU/liter and normal free T(4)) participated in the study. Each patient normalized their TSH values without T(4) therapy throughout the follow-up. MEASUREMENTS: TSH and free T(4) levels were evaluated every 6 months. RESULTS: Normalization occurred at a median time of 18 months (range, 6-60 months). Fifteen patients normalized their TSH levels during the first year of follow-up and 27 during the first 2 yr. Ten patients normalized their TSH values at the fourth or fifth year. Only four patients reverted to TSH values less than 2 mU/liter. Final TSH levels achieved by the patients were significantly correlated with the time elapsed until normalizing these levels (r = 0.367; P = 0.020). CONCLUSIONS: There is no clear pattern of TSH normalization, although most patients normalize their TSH values early in the follow-up. The final TSH reached seems to be related to the time of normalization. PMID- 15811926 TI - Joint mechanisms of impaired growth-hormone pulse renewal in aging men. AB - CONTEXT: Aging reduces the size (mass) of GH secretory bursts and thereby reduces total GH secretion. Experimental data indicate that high-amplitude GH pulses are evoked by reversible cycles of GH-induced negative feedback. Whether aging impairs autofeedback is unknown. OBJECTIVE: The objective of this study is to assess whether age attenuates and IGF-I potentiates negative feedback by a near physiological pulse of GH. DESIGN/SETTING/SUBJECTS: In a university setting, 17 healthy men ages 19-71 yr each underwent four randomly ordered infusion studies on separate mornings fasting. INTERVENTION: Intravenous injection of a pulse of: 1) saline or 2) recombinant human (rh) GH to impose controlled negative feedback, followed in 2 h by a bolus of 3) saline or (iv) the ghrelin analog GHRP-2 to overcome feedback inhibition. OUTCOME MEASURES: The impact of age and IGF-I concentrations on GH autofeedback was assessed by regression analysis. RESULTS: Percentage feedback inhibition correlated negatively with: 1) age after consecutive rh GH/saline infusion (R(2) = 0.42, P = 0.005) at any IGF-I concentration; and 2) total IGF-I concentrations after rh GH/GHRP-2 infusion (R(2) = 0.40, P = 0.009) at any age. In contrast, sex-steroid concentrations and body mass index were unrelated to degree of autoinhibition. CONCLUSIONS: Increased age in healthy men predicts impaired GH autofeedback, which may contribute to attenuated renewal of high-amplitude GH pulses. Conversely, higher IGF-I concentrations in young men forecast accentuated GH autoinhibition, which may drive prominent GH pulses. PMID- 15811927 TI - Hyperinsulinemic hypoglycemia in Beckwith-Wiedemann syndrome due to defects in the function of pancreatic beta-cell adenosine triphosphate-sensitive potassium channels. AB - BACKGROUND: Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth syndrome that is clinically and genetically heterogeneous. Hyperinsulinemic hypoglycemia occurs in about 50% of children with BWS and, in the majority of infants, it resolves spontaneously. However, in a small group of patients the hypoglycemia can be persistent and may require pancreatectomy. The mechanism of persistent hyperinsulinemic hypoglycemia in this group of patients is unclear. PATIENTS AND METHODS: Using patch-clamp techniques on pancreatic tissue obtained at the time of surgery, we investigated the electrophysiological properties of ATP-sensitive K(+) (K(ATP)) channels in pancreatic beta-cells in a patient with BWS and severe medically-unresponsive hyperinsulinemic hypoglycemia. RESULTS: Persistent hyperinsulinism was found to be caused by abnormalities in K(ATP) channels of the pancreatic beta-cell. Immunofluorescence studies using a SUR1 antibody revealed perinuclear pattern of staining in the BWS cells, suggesting a trafficking defect of the SUR1 protein. No mutations were found in the genes ABCC8 and KCNJ11 encoding for the two subunits, SUR1 and KIR6.2, respectively, of the K(ATP) channel. Genetic analysis of this patients BWS showed evidence of mosaic paternal isodisomy. CONCLUSIONS: In this novel case of BWS with mosaic paternal uniparental disomy for 11p15, persistent hyperinsulinism was due to abnormalities in K(ATP) channels of the pancreatic beta-cell. The mechanism/s by which mosaic paternal uniparental disomy for 11p15 causes a trafficking defect in the SUR1 protein of the K(ATP) channel remains to be elucidated. PMID- 15811928 TI - Obesity and type 2 diabetes do not alter splanchnic cortisol production in humans. AB - CONTEXT: Cortisol is a potent regulator of carbohydrate, fat, and protein metabolism. OBJECTIVE: The objective of the study was to determine whether obesity alone or in combination with type 2 diabetes increases splanchnic and/or leg cortisol production. DESIGN: Splanchnic and leg cortisol production were measured using the hepatic and leg catheterization technique combined with infusion of D4-cortisol. SETTING: The study was conducted in a General Clinical Research Center. PARTICIPANTS: Nine lean nondiabetic, 10 obese nondiabetic, and 11 obese diabetic subjects were studied. INTERVENTIONS: Diabetic volunteers were withdrawn from their glucose-lowering medications before study. MAIN OUTCOME MEASURES: Rates of total body, splanchnic and leg cortisol, and D3-cortisol production were measured. RESULTS: Rates of splanchnic cortisol production equaled or exceeded those occurring in extrasplanchnic tissues (e.g. the adrenals) in all three groups. However, because concurrent splanchnic cortisol uptake also occurred, net splanchnic cortisol release was minimal. Splanchnic cortisol production and splanchnic D3-cortisol production (an index of splanchnic 11beta-hydroxysteroid dehydrogenase type 1 activity) did not differ among the three groups. In addition, splanchnic cortisol production did not correlate with either visceral fat or endogenous glucose production. On the other hand, splanchnic cortisol uptake was greater in the obese diabetic than lean nondiabetic subjects (25 +/- 2.9 vs. 15.3 +/- 2.5 microg/min; P < 0.05). Splanchnic, but not leg, D3-cortisol production was correlated with total body D3 cortisol production (r = 0.70; P < 0.001). CONCLUSIONS: Although large amounts of cortisol are produced within the splanchnic bed, implying high intrahepatic glucocorticoid concentrations, rates do not differ in lean and obese nondiabetic humans and are not influenced by the presence of type 2 diabetes mellitus. On the other hand, obesity but not diabetes increases splanchnic cortisol uptake. PMID- 15811929 TI - Long-term follow-up of 106 multiple sclerosis patients undergoing interferon-beta 1a or 1b therapy: predictive factors of thyroid disease development and duration. AB - BACKGROUND: Conflicting data have been reported on the association between interferon (IFN)-beta therapy of multiple sclerosis (MS) patients and thyroid disease development. AIMS: The goals of this study are as follows: to assess the actual occurrence of thyroid dysfunction and autoimmunity during long-term IFN beta therapy; to establish the possible presence of predictive factors for thyroid dysfunction development and duration; and to suggest an effective follow up protocol for patients receiving long-term IFN-beta therapy. STUDY PROTOCOL: A total of 106 MS patients (76 women) underwent IFN-beta 1a or 1b therapy for up to 84 months (median, 42 months). Thyroid function and autoimmunity were assessed at baseline and every 3-6 months throughout the treatment course. RESULTS: Baseline thyroid autoimmunity was detected in 8.5% of patients and hypothyroidism in 2.8%. Thyroid dysfunction (80% hypothyroidism, 92% subclinical, 56% transient) developed in 24% (68% with autoimmunity) of patients and autoimmunity in 22.7% (45.5% with dysfunction), without significant differences between the two cytokines; 68% of dysfunctions occurred within the first year. Autoimmunity emerged as the only predictive factor for dysfunction development (relative risk, 8.9), whereas sustained disease was significantly associated with male gender (P < 0.003). CONCLUSIONS: Both incident thyroid autoimmunity and dysfunction frequently occur in MS patients during IFN-beta therapy, particularly within the first year of treatment. Thyroid dysfunction is generally subclinical and transient in over than half of cases; preexisting or incident autoimmunity emerged as the only significant predictive factor for thyroid dysfunction development. Thyroid function and autoimmunity assessment is mandatory within the first year of IFN-beta therapy; thereafter, serum TSH measurement only in patients with thyroid disease could be sufficient. PMID- 15811930 TI - Combined inhibition of types I and II 5 alpha-reductase selectively augments the basal (nonpulsatile) mode of testosterone secretion in young men. AB - CONTEXT: Testosterone (Te) is metabolized in the hypothalamus and pituitary gland, where untransformed steroid and activated products participate in feedback regulation of GnRH and LH secretion. Genetic inactivation of 5 alpha-reductase type I remains undescribed clinically, whereas deficiency of the type II isoenzyme elevates both LH and Te concentrations. OBJECTIVE: The aim of this study was to test the combined feedback contribution of 5 alpha-reduced steroids. SETTING/DESIGN/INTERVENTION: In a university setting, nine young men received placebo and a dual (type I/type II) 5 alpha-reductase inhibitor, dutasteride. METHODS/OUTCOMES: LH and Te dynamics were assessed by: 1) 10-min blood sampling for 26 h; 2) GnRH stimulation (100 ng/kg iv); 3) discrete peak detection; 4) deconvolution analysis; 5) cosinor analyses of 24-h rhythmicity; and 6) pattern regularity. RESULTS: Compared with placebo, dutasteride lowered 5 alpha-dihydro Te concentrations by 80% (P = 0.009), but did not alter any measure of LH dynamics. Conversely, dutasteride augmented: 1) total, bioavailable and free Te concentrations (0.002 < P < 0.032) without changing estradiol or SHBG concentrations; 2) nadir Te concentrations (P = 0.025); and 3) basal (P = 0.013) and thereby total (basal plus pulsatile) (P = 0.003) Te secretion. CONCLUSION: Combined antagonism of types I and II 5 alpha-reductase preferentially drives nonpulsatile Te secretion in healthy men. The concomitant stability of LH outflow could indicate that intragonadal 5 alpha-reduced androgens repress basal Leydig cell steroidogenesis. PMID- 15811931 TI - The impact on clinical practice of routine screening for macroprolactin. AB - BACKGROUND: Macroprolactin has reduced bioactivity in vivo and accumulates in the sera of some subjects, resulting in pseudo-hyperprolactinemia and consequent misdiagnosis. METHODS: We have audited our experience of routine screening for macroprolactin using polyethylene glycol (PEG) precipitation over a 5-yr period in a single center. RESULTS: Application of a reference range for monomeric prolactin (the residual prolactin present in macroprolactin-depleted serum) for normal individuals revealed that 453 of 2089 hyperprolactinemic samples (22%) identified by Delfia immunoassay were explained entirely by macroprolactin. The percentage of hyperprolactinemic samples explained by macroprolactinemia was similar across all levels of total prolactin (18, 21, 19, and 17% of samples from 700-1000, 1000-2000, 2000-3000, and greater than 3000 mU/liter, respectively). Application of an absolute prolactin threshold after polyethylene glycol treatment of sera, rather than the traditional method, i.e. less than 40% recovery, minimizes the opportunity for misclassification of patients in whom macroprolactin accounted for more than 60% of prolactin and the residual bioactive prolactin was present in excess. Macroprolactinemic patients could not be differentiated from true hyperprolactinemic patients on the basis of clinical features alone. Although oligomenorrhea/amenorrhea and galactorrhea were more common in patients with true hyperprolactinemia (P < 0.05), they were also frequently present in macroprolactinemic patients. Plasma levels of estradiol and LH and the LH/FSH ratio were significantly greater in macroprolactinemic compared with true hyperprolactinemic subjects (P < 0.05). Reduced use of imaging and dopamine agonist treatment resulted in a net cost savings, offsetting the additional cost associated with the introduction of screening. CONCLUSION: Routine screening of all hyperprolactinemic sera for macroprolactin is recommended. PMID- 15811932 TI - A potential rearrangement between CYP19 and TRPM7 genes on chromosome 15q21.2 as a cause of aromatase excess syndrome. AB - CONTEXT: Aromatase excess syndrome (AES) is a rare hereditary autosomal dominant disorder characterized by increased extraglandular aromatization of steroids and presented with heterosexual precocity in males and isosexual precocity in females. OBJECTIVE: The objective was to study the molecular basis of AES in a kindred with 16 affected subjects, both males and females. PATIENTS: The propositus, currently a 17-year-old boy, presented with breast enlargement in the first year of life, which persisted thereafter. Investigations at the age of 7.5 yr revealed growth acceleration (height sd score, 2.8), puberty staging Tanner P1B3, testicular volume 6 ml, and bone age 13 yr. The hormonal data were compatible with increased conversion of androgens to estrogens, which was independent of gonadotropin secretion. In the affected adults, there were short stature (height sd score ranged from -3.7 to -2), gynecomastia in males, and macromastia in females. DESIGN: Linkage analysis was performed using a polymorphic tetranucleotide (TTTA) repeat marker at nucleotide position 682 of CYP gene, as well as two additional STS markers, D15S123 (CA)n and D15S209 (CA)n, located within genetic distance of less than 5 cM from CYP19 gene. Using RNA extracted from the breast tissue of the propositus, a 5'-rapid amplification of cDNA ends (RACE) was performed with gene-specific primers corresponding to exon 2 of CYP19 gene. RESULTS: Linkage analysis with (TTTA)n, D15S123 (CA)n, and D15S209 (CA)n markers produced LOD scores 0.85, 1.5, and 1.17, respectively. 5'-RACE revealed a novel untranslated exon 1 composed by exon 1 of TRPM7 gene (Transient Receptor Potential Cation Channel, Subfamily M, member 7), which has ubiquitous expression. CONCLUSIONS: 5'-RACE finding points to a potential rearrangement between CYP19 and TRPM7 genes on chromosome 15q21.2 as a cause of AES. PMID- 15811934 TI - Primary adrenal insufficiency in children: twenty years experience at the Sainte Justine Hospital, Montreal. AB - Primary adrenal insufficiency (PAI) in the pediatric population (0-18 yr) is most commonly attributed to congenital adrenal hyperplasia (CAH), which occurs in about 1 in 15,000 births, followed by Addison's disease, with an assumed autoimmune etiology. However, molecular advances have increased the number of possible diagnoses. The objective of this study was to determine the incidence and etiologies of PAI in our pediatric population. All patients with a diagnosis of PAI followed by the Endocrinology Service at our institution between September 1981 and September 2001 were studied. One hundred three patients (48 boys) were identified, primarily by the Endocrinology Clinic case registry. CAH was the most frequent etiology (71.8%). However, non-CAH etiologies accounted for 28.2%, of which 55% were nonautoimmune in etiology. Importantly, the CAH sex ratio was 1:1, despite the absence of biochemical screening for this condition in Quebec newborns. Patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dysplasia (APECED) developed adrenal insufficiency 4 yr earlier than those with non-autoimmune disease. Finally, we review the rare etiologies of PAI and propose an algorithm to aid in targeted genetic testing. PMID- 15811933 TI - Autosomal dominant neurohypophyseal diabetes insipidus with linkage to chromosome 20p13 but without mutations in the AVP-NPII gene. AB - CONTEXT: Autosomal dominant neurohypophyseal diabetes insipidus (ADNDI) has been known as a rare disorder transmitted as an autosomal dominant trait, characterized by polyuria and polydipsia, and caused by deficient neurosecretion of arginine vasopressin precursor (AVP-NPII). We reported an ADNDI family with linkage to chromosome 20p13 but without mutations in the AVP-NPII gene. OBJECTIVE: The objective of this study was to identify the corresponding locus responsible for ADNDI in a family without AVP-NP II gene mutations. SUBJECTS AND METHODS: Two families with ADNDI were diagnosed by water deprivation test. The AVP-NPII gene was amplified by PCR and sequenced. A genomewide scan was performed in one family using 400 microsatellite markers covering 22 autosomes. RESULTS: A 3-bp deletion (1827-1829delAGG) of AVP-NPII gene was identified in the affected individuals in one family. Although no mutations could be detected in the coding, the promoter, and intronic regions of AVP-NPII gene in the other family, a maximum LOD score of 1.202999 (theta = 0.00) was obtained at marker D20S889 by genomewide scan, and a 7-cM interval on chromosome 20p13 was defined by fine mapping with markers D20S199-D20S849. Furthermore, the intragenic region that regulates AVP-NPII and oxytocin expression as an enhancer element and the UBCE7IP5 gene that participates in prohormone degradation were sequenced. No alterations could be detected either. CONCLUSION: The corresponding locus responsible for ADNDI is possibly heterogeneous regarding the slightly different clinical features in these two families. PMID- 15811935 TI - Up-regulation of gap junctional intercellular communication and connexin43 expression by retinoic acid in human endometrial stromal cells. AB - CONTEXT: Gap junctions, made up of connexins (Cxs), play fundamental roles in coordinating a number of cellular processes through their ability to directly regulate cell-cell communication. Cx43 is the most widely expressed Cx in the endometrium and is known to be important in a variety of physiological and pathological processes in this tissue. OBJECTIVE: In this study, we investigated the ability of the retinoid, all-trans-retinoic acid (RA), to regulate Cx43 expression in human endometrial stromal cells. DESIGN: Primary endometrial stromal cells obtained from patients undegoing surgery for infertility workup were treated in vitro with RA and control compounds for different time periods, up to 48 h. Cx43 mRNA and protein levels, protein phosphorylation, and gap junctional intercellular communication (GJIC) were analyzed. RESULTS: Treatment of the cells with RA showed a dose-dependent increase in Cx43 expression at both the mRNA and protein levels. In addition, RA induced a relative decrease in the phosphorylated species of Cx43 while causing a corresponding increase in the nonphosphorylated form. Concomitant with these changes, RA-treated cells demonstrated up to a 250% enhancement of GJIC as assessed by dye transfer experiments. Augmentation of GJIC and alterations of Cx43 expression were observed over the same range of RA concentrations. Treatment of cells with the protein kinase C activator 12-O-tetradecanoylphorbol-13-acetate increased the phosphorylated species of Cx43 and correspondingly inhibited GJIC. CONCLUSIONS: Phosphorylation of Cx43 is inversely related to GJIC in endometrial stromal cells. Retinoids increase GJIC in endomentrial stromal cells through upregulation of Cx43 expression while inducing a decrease in the phosphorylated species of the protein. The data suggest a novel mechanism by which retinoids can influence endometrial cell biology. PMID- 15811936 TI - Histone modifications: combinatorial complexity or cumulative simplicity? PMID- 15811937 TI - CcbP, a calcium-binding protein from Anabaena sp. PCC 7120, provides evidence that calcium ions regulate heterocyst differentiation. AB - Although it is known that calcium is a very important messenger involved in many eukaryotic cellular processes, much less is known about calcium's role in bacteria. CcbP, a Ca(2+)-binding protein, was isolated from the heterocystous cyanobacterium Anabaena sp. PCC 7120, and the ccbP gene was cloned and inactivated. In the absence of combined nitrogen, inactivation of ccbP resulted in multiple contiguous heterocysts, whereas overexpression of ccbP suppressed heterocyst formation. Calmodulin, which is not present in Anabaena species, could also suppress heterocyst formation in both Anabaena sp. PCC 7120 and Anabaena variabilis. HetR induction upon nitrogen step-down was slow in the strain overexpressing ccbP. The Ca(2+) reporter protein obelin was used to show that mature heterocysts had a high intracellular free Ca(2+)concentration {[Ca(2+)](i)}, and immunoblotting showed that CcbP was absent from heterocysts. A regular pattern of cells with higher [Ca(2+)](i) was established during heterocyst differentiation before the appearance of proheterocysts. A rapid increase of [Ca(2+)](i) could be detected 4 h after the removal of combined nitrogen, and this increase was suppressed by excessive CcbP. These results suggest that Ca(2+) ions play very important roles in hetR induction and heterocyst differentiation. PMID- 15811938 TI - Biography of Veerabhadran Ramanathan. PMID- 15811939 TI - Big brains do matter in new environments. PMID- 15811940 TI - Threshold effects in gene regulation: when some is not enough. PMID- 15811942 TI - From the representation theory of vertex operator algebras to modular tensor categories in conformal field theory. PMID- 15811941 TI - RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model. AB - Huntington's disease (HD) is a fatal, dominant neurogenetic disorder. HD results from polyglutamine repeat expansion (CAG codon, Q) in exon 1 of HD, conferring a toxic gain of function on the protein huntingtin (htt). Currently, no preventative treatment exists for HD. RNA interference (RNAi) has emerged as a potential therapeutic tool for treating dominant diseases by directly reducing disease gene expression. Here, we show that RNAi directed against mutant human htt reduced htt mRNA and protein expression in cell culture and in HD mouse brain. Importantly, htt gene silencing improved behavioral and neuropathological abnormalities associated with HD. Our data provide support for the further development of RNAi for HD therapy. PMID- 15811943 TI - Biography of George W. Flynn. PMID- 15811944 TI - Directed evolution of specific receptor-ligand pairs for use in the creation of gene switches. AB - Despite their versatility and power in controlling gene regulation in nature, nuclear hormone receptors (NHRs) have largely eluded utility in heterologous gene regulation applications such as gene therapy and metabolic engineering. The main reason for this void is the pleiotropic interference of the receptor-ligand combination with regulatory networks in the host organism. In recent years, numerous strategies have been developed to engineer ligand-receptor pairs that do not cross-interact with host regulatory pathways. However, these strategies have either met with limited success or cannot be readily extended to other ligand receptor pairs. Here, we present a simple, effective, and readily generalizable strategy for reengineering NHRs to respond specifically to a selected synthetic ligand. The method involves generation of genetic diversity by stepwise individual site saturation mutagenesis of a fixed set of ligand-contacting residues and random point mutagenesis, followed by phenotypic screening based on a yeast two-hybrid system. As a test case, this method was used to alter the specificity of the NHR human estrogen receptor alpha in favor of the synthetic ligand 4,4'-dihydroxybenzil, relative to the natural ligand 17beta-estradiol, by >10(7)-fold. The resulting ligand-receptor pair is highly sensitive to the synthetic ligand in human endometrial cancer cells and is essentially fully orthogonal to the wild-type receptor-natural ligand pair. This method should provide a powerful, broadly applicable tool for engineering receptors/enzymes with improved or novel ligand/substrate specificity. PMID- 15811945 TI - Impact of distal mutations on the network of coupled motions correlated to hydride transfer in dihydrofolate reductase. AB - A comprehensive analysis of the network of coupled motions correlated to hydride transfer in dihydrofolate reductase is presented. Hybrid quantum/classical molecular dynamics simulations are combined with a rank correlation analysis method to extract thermally averaged properties that vary along the collective reaction coordinate according to a prescribed target model. Coupled motions correlated to hydride transfer are identified throughout the enzyme. Calculations for wild-type dihydrofolate reductase and a triple mutant, along with the associated single and double mutants, indicate that each enzyme system samples a unique distribution of coupled motions correlated to hydride transfer. These coupled motions provide an explanation for the experimentally measured nonadditivity effects in the hydride transfer rates for these mutants. This analysis illustrates that mutations distal to the active site can introduce nonlocal structural perturbations and significantly impact the catalytic rate by altering the conformational motions of the entire enzyme and the probability of sampling conformations conducive to the catalyzed reaction. PMID- 15811946 TI - Identification of the control region for tissue-specific imprinting of the stimulatory G protein alpha-subunit. AB - Gnas is a complex gene with multiple imprinted promoters. The upstream Nesp and Nespas/Gnasxl promoters are paternally and maternally methylated, respectively. The downstream promoter for the stimulatory G protein alpha-subunit (G(s)alpha) is unmethylated, although in some tissues (e.g., renal proximal tubules), G(s)alpha is poorly expressed from the paternal allele. Just upstream of the G(s)alpha promoter is a primary imprint mark (1A region) where maternal-specific methylation is established during oogenesis. Pseudohypoparathyroidism type 1B, a disorder of renal parathyroid hormone resistance, is associated with loss of 1A methylation. Analysis of embryos of Dnmt3L(-/-) mothers (which cannot methylate maternal imprint marks) showed that Nesp, Nespas/Gnasxl, and 1A imprinting depend on one or more maternal primary imprint marks. We generated mice with deletion of the 1A differentially methylated region. These mice had normal Nesp-Nespas/Gnasxl imprinting, indicating that the Gnas locus contains two independent imprinting domains (Nespas-Nespas/Gnasxl and 1A-G(s)alpha) controlled by distinct maternal primary imprint marks. Paternal, but not maternal, 1A deletion resulted in G(s)alpha overexpression in proximal tubules and evidence for increased parathyroid hormone sensitivity but had no effect on G(s)alpha expression in other tissues where G(s)alpha is normally not imprinted. The 1A region is a maternal imprint mark that contains one or more methylation-sensitive cis-acting elements that suppress G(s)alpha expression from the paternal allele in a tissue specific manner. PMID- 15811947 TI - Cleavage furrow formation and ingression during animal cytokinesis: a microtubule legacy. AB - Cytokinesis ensures the proper partitioning of the nuclear and cytoplasmic contents into independent daughter cells at the end of cell division. Although the metazoan mitotic spindle has been implicated in the placement and advancement of the cleavage furrow, the molecules responsible for these processes have remained elusive. Recent studies have provided insights into the role of different microtubule structures and associated proteins in cleavage furrow positioning and ingression together with the signalling events that regulate the dynamics of the equatorial cell cortex during cytokinesis. We try to unify these findings into a general model of cytokinesis in which both astral and central spindle microtubules have the ability to induce furrowing. We further propose that the evolutionarily conserved centralspindlin complex serves as a master controller of cell cleavage in Drosophila by promoting both furrow formation and ingression. The same mechanism might be conserved in other organisms. PMID- 15811948 TI - Multi-lineage potential of fetal cells in maternal tissue: a legacy in reverse. AB - Fetal cells circulate in pregnant women and persist in blood and tissue for decades post-partum. The mother thus becomes chimeric. Factors that may influence such fetal cell microchimerism include histocompatibility, fetal or placental abnormalities, or a reproductive history that includes miscarriage or elective termination. Fetal cell microchimerism is associated with some maternal autoimmune diseases, such as systemic sclerosis. Moreover, a novel population of fetal cells, the pregnancy-associated progenitor cells (PAPCs), appears to differentiate in diseased or injured maternal tissue. The cellular origin of these cells is at present unknown but could be a hematopoietic stem cell, a mesenchymal stem cell, or a novel cell type. Pregnancy therefore results in the acquisition of cells with stem-cell-like properties that may influence maternal health post-partum. Rather than triggering disease, these cells may instead combat it. PMID- 15811949 TI - Secretory pathway Ca(2+)-ATPase (SPCA1) Ca(2)+ pumps, not SERCAs, regulate complex [Ca(2+)](i) signals in human spermatozoa. AB - The sarcoplasmic-endoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitors thapsigargin (0.1-1 microM) and cyclopiazonic acid (10 microM), failed to affect resting [Ca(2+)] in human spermatozoa. Slow progesterone-induced [Ca(2+ i)](i) oscillations in human spermatozoa, which involve cyclic emptying-refilling of an intracellular Ca(2+) store were also insensitive to these inhibitors. Non selective doses of thapsigargin (5-30 microM, 50-300 times the saturating dose for SERCA inhibition), caused elevation of resting [Ca(2+)](i) and partial, dose dependent disruption of oscillations. A 10-40 microM concentration of bis(2 hydroxy-3-tert-butyl-5-methyl-phenyl)methane (bis-phenol), which inhibits both thapsigargin-sensitive and -insensitive microsomal Ca(2+) ATPases, caused elevation of resting [Ca(2+)](i) and inhibition of [Ca(2+)](i) oscillations at doses consistent with inhibition of thapsigargin-resistant, microsomal ATPase and liberation of stored Ca(2+). Low doses of bis-phenol had marked effects on [Ca(2+)](i) oscillation kinetics. Application of the drug to cells previously stimulated with progesterone had effects very similar to those observed when it was applied to unstimulated cells, suggesting that the sustained Ca(2+) influx induced by progesterone is not mediated via mobilisation of Ca(2+) stores. Western blotting for human sperm proteins showed expression of secretory pathway Ca(2+) ATPase (SPCA1). Immunolocalisation studies revealed expression of SPCA1 in all cells in an area behind the nucleus, extending into the midpiece. Staining for SERCA, carried out in parallel, detected no expression with either technique. We conclude that: (1) intracellular Ca(2+) store(s) and store-dependent [Ca(2+)](i) oscillations in human spermatozoa rely primarily on a thapsigargin/cyclopiazonic acid-insensitive Ca(2+) pump, which is not a SERCA as characterised in somatic cells; (2) effects of high-dose thapsigargin on spermatozoa primarily reflect non-specific actions on non-SERCAs and; (3) secretory pathway Ca(2+) ATPases contribute at least part of this non-SERCA Ca(2+) pump activity. PMID- 15811950 TI - DCC association with lipid rafts is required for netrin-1-mediated axon guidance. AB - During development, axons migrate long distances in responses to attractive or repulsive signals that are detected by their growth cones. One of these signals is mediated by netrin-1, a diffusible laminin-related molecule that both attracts and repels growth cones via interaction with its receptor DCC (deleted in colorectal cancer). Here we show that DCC in both commissural neurons and immortalized cells, is partially associated with cholesterol- and sphingolipid enriched membrane domains named lipid rafts. This localization of DCC in lipid rafts is mediated by the palmitoylation within its transmembrane region. Moreover, this raft localization of DCC is required for netrin-1-induced DCC dependent ERK activation, and netrin-1-mediated axon outgrowth requires lipid raft integrity. Thus, the presence of axon guidance-related receptors in lipid rafts appears to be a crucial pre-requisite for growth cone response to chemo attractive or repulsive cues. PMID- 15811951 TI - Ocular surface epithelia contain ABCG2-dependent side population cells exhibiting features associated with stem cells. AB - When cell populations are incubated with the DNA-binding dye Hoechst 33342 and subjected to flow cytometry analysis for Hoechst 33342 emissions, active efflux of the dye by the ABCG2/BCRP1 transporter causes certain cells to appear as a segregated cohort, known as a side population (SP). Stem cells from several tissues have been shown to possess the SP phenotype. As the lack of specific surface markers has hindered the isolation and subsequent biochemical characterization of epithelial stem cells this study sought to determine the existence of SP cells and expression of ABCG2 in the epithelia of the ocular surface and evaluate whether such SP cells had features associated with epithelial stem cells. Human and rabbit limbal-corneal and conjunctival epithelial cells were incubated with Hoechst 33342, and analyzed and sorted by flow cytometry. Sorted cells were subjected to several tests to determine whether the isolated SP cells displayed features consistent with the stem cell phenotype. Side populations amounting to <1% of total cells, which were sensitive to the ABCG2-inhibitor fumitremorgin C, were found in the conjunctival and limbal epithelia, but were absent from the stem cell-free corneal epithelium. Immunohistochemistry was used to establish the spatial expression pattern of ABCG2. The antigen was detected in clusters of conjunctival and limbal epithelia basal cells but was not present in the corneal epithelium. SP cells were characterized by extremely low light side scattering and contained a high percentage of cells that: showed slow cycling prior to tissue collection; exhibited an initial delay in proliferation after culturing; and displayed clonogenic capacity and resistance to phorbol-induced differentiation; all features that are consistent with a stem cell phenotype. PMID- 15811952 TI - The tumor suppressor TSLC1/NECL-2 triggers NK-cell and CD8+ T-cell responses through the cell-surface receptor CRTAM. AB - The tumor suppressor in lung cancer-1 (TSLC1) gene is frequently silenced in human lung carcinomas, and its expression suppresses tumorigenesis in nude mice. TSLC1 encodes a cell-surface protein called Necl-2 that belongs to the Nectin and Nectin-like (Necl) family of molecules. Necl-2 mediates epithelial cell junctions by homotypic contacts and/or heterotypic interactions with other Nectins and Necls. Thus, it inhibits tumorigenesis by ensuring that epithelial cells grow in organized layers. Here, we demonstrate that natural killer (NK) cells and CD8+ T cells recognize Necl-2 through a receptor known as class I-restricted T-cell associated molecule (CRTAM), which is expressed only on activated cells. CRTAM Necl-2 interactions promote cytotoxicity of NK cells and interferon gamma (IFN gamma) secretion of CD8+ T cells in vitro as well as NK cell-mediated rejection of tumors expressing Necl-2 in vivo. These results provide evidence for an additional mechanism of tumor suppression mediated by TSLC1 that involves cytotoxic lymphocytes. Furthermore, they reveal Necl-2 as one of the molecular targets that allows the immunosurveillance network to distinguish tumor cells from normal cells. PMID- 15811953 TI - Low-molecular-weight heparins for thromboprophylaxis and treatment of venous thromboembolism in pregnancy: a systematic review of safety and efficacy. AB - To assess the safety and efficacy of low-molecular-weight heparins (LMWHs) for thromboprophylaxis and treatment of venous thromboembolism (VTE) in pregnancy, a systematic review of studies to the end of 2003 was undertaken. Data on VTE recurrence and side effects were extracted and cumulative incidences of VTE and adverse effects calculated. Of 81 reports identified, 64 reporting 2777 pregnancies were included. In 15 studies (174 patients) the indication for LMWH was treatment of acute VTE, and in 61 studies (2603 pregnancies) it was thromboprophylaxis or adverse pregnancy outcome. There were no maternal deaths. VTE and arterial thrombosis (associated with anti-phospholipid syndrome) were reported in 0.86% (95% confidence interval [CI], 0.55%-1.28%) and 0.50% (95% CI, 0.28%-0.84%) of pregnancies, respectively. Significant bleeding, generally associated with primary obstetric causes, occurred in 1.98% (95% CI, 1.50% 2.57%), allergic skin reactions in 1.80% (95% CI, 1.34%-2.37%), heparin-induced thrombocytopenia in 0%, thrombocytopenia (unrelated to LMWH) in 0.11% (95% CI, 0.02%-0.32%), and osteoporotic fracture in 0.04% (95% CI, < 0.01%-0.20%) of pregnancies. Overall, live births were reported in 94.7% of pregnancies, including 85.4% in those receiving LMWH for recurrent pregnancy loss. LMWH is both safe and effective to prevent or treat VTE in pregnancy. PMID- 15811954 TI - Zygotic nucleosome assembly protein-like 1 has a specific, non-cell autonomous role in hematopoiesis. AB - Nucleosome assembly proteins (NAPs) bind core histones, facilitate chromatin remodeling, and can act as transcriptional coactivators. We previously described the isolation of a Xenopus NAP1-like (xNAP1L) cDNA, which encodes a member of this protein family. Its zygotic expression is restricted to neural cells, the outer cells of the ventral blood island (VBIs), and the ectoderm overlying the blood precursors. Here, we report that depletion of zygotic xNAP1L in embryos produces no obvious morphologic phenotype, but ablates alpha-globin mRNA expression in the VBIs. Transcript levels of the hematopoietic precursor genes SCL and Xaml (Runx-1) are also reduced in the VBIs. SCL expression can be rescued by injection of xNAP1L mRNA into the ectoderm, showing that the effect of xNAP1L can be non-cell autonomous. Fli1 and Hex, genes expressed in hemangioblasts but subsequently endothelial markers, were unaffected, suggesting that xNAP1L is required for the hematopoietic lineage specifically. Our data are consistent with a requirement for xNAP1L upstream of SCL, and injection of SCL mRNA into xNAP1L depleted embryos rescues alpha-globin expression. Thus, xNAP1L, which belongs to a family of proteins previously believed to have general roles, has a specific function in hematopoiesis. PMID- 15811955 TI - Familial risk for non-Hodgkin lymphoma and other lymphoproliferative malignancies by histopathologic subtype: the Swedish Family-Cancer Database. AB - Non-Hodgkin lymphoma (NHL) consists of a heterogeneous group of tumors. Population-based data on the familial risk for specific histopathologic subtypes have not been established. Such data are useful for clinical counseling and for searching tumor subtypes sharing common genetic pathways. We used the Swedish Family-Cancer Database to calculate standardized incidence ratios (SIRs) for histopathology-specific subtypes of NHL in 4455 offspring with NHL whose parents or siblings were affected with different types of lymphoproliferative malignancies. A familial history of NHL significantly increased the risk for NHL (SIRparent = 1.8; SIRsibling = 1.9) and for diffuse large B-cell lymphoma (SIRparent = 2.3), follicular lymphoma (SIRsibling = 2.3), and B-cell lymphoma not otherwise specified (NOS) (SIRsibling = 3.4). For a parental history of histopathology-specific concordant cancer, the risks were significantly increased for diffuse large B-cell lymphoma (SIR = 11.8), follicular NHL (SIR = 6.1), plasma cell myeloma (SIR = 2.5), and chronic lymphocytic leukemia (SIR = 5.9). Familial clusters for NHL seemed stronger in females and in siblings. Our study provides the first quantification of the familial risks for NHL by histopathology. The present findings give evidence for a strong familial association of NHL, with little differences in the magnitude of risks for various histopathologic subtypes. The patterns of risks in parents and siblings support the hypothesis of an autosomal-dominant component for diffuse large B-cell NHL and a recessive one for follicular NHL. PMID- 15811956 TI - ICAM-1 regulates neutrophil adhesion and transcellular migration of TNF-alpha activated vascular endothelium under flow. AB - In vivo, leukocyte transendothelial migration (TEM) occurs at endothelial cell junctions (paracellular) and nonjunctional (transcellular) locations, whereas in vitro models report that TEM is mostly paracellular. The mechanisms that control the route of leukocyte TEM remain unknown. Here we tested the hypothesis that elevated intercellular adhesion molecule-1 (ICAM-1) expression regulates the location of polymorphonuclear leukocyte (PMN) TEM. We used an in vitro flow model of tumor necrosis factor-alpha (TNF-alpha)-activated human umbilical vein endothelium cells (HUVECs) or an HUVEC cell line transfected with ICAM-1GFP (green fluorescent protein) and live-cell fluorescence microscopy to quantify the location of PMN adhesion and TEM. We observed robust transcellular TEM with TNF alpha-activated HUVECs and ICAM-1GFP immortalized HUVECS (iHUVECs). In contrast, primary CD3+ T lymphocytes exclusively used a paracellular route. Endothelial ICAM-1 was identified as essential for both paracellular and transcellular PMN transmigration, and interfering with ICAM-1 cytoplasmic tail function preferentially reduced transcellular TEM. We also found that ICAM-1 surface density and distribution as well as endothelial cell shape contributed to transcellular TEM. In summary, ICAM-1 promotes junctional and nonjunctional TEM across inflamed vascular endothelium via distinct cytoplasmic tail associations. PMID- 15811957 TI - Molecular mechanism and functional implications of thrombin-mediated tyrosine phosphorylation of PKCdelta in platelets. AB - Thrombin has been known to cause tyrosine phosphorylation of protein kinase C delta (PKCdelta) in platelets, but the molecular mechanisms and function of this tyrosine phosphorylation is not known. In this study, we investigated the signaling pathways used by protease-activated receptors (PARs) to cause tyrosine phosphorylation of PKCdelta and the role of this event in platelet function. PKCdelta was tyrosine phosphorylated by either PAR1 or PAR4 in a concentration- and time-dependent manner in human platelets. In particular, the tyrosine 311 residue was phosphorylated downstream of PAR receptors. Also the tyrosine phosphorylation of PKCdelta did not occur in Galpha(q)-deficient mouse platelets and was inhibited in the presence of a phospholipase C (PLC) inhibitor U73122 and calcium chelator BAPTA (5,5'-dimethyl-bis(o-aminophenoxy)ethane-N, N, N ', N ' tetraacetic acid), suggesting a role for Galpha(q) pathways and calcium in this event. Both PAR1 and PAR4 caused a time-dependent activation of Src (pp60c-src) tyrosine kinase and Src tyrosine kinase inhibitors completely blocked the tyrosine phosphorylation of PKCdelta. Inhibition of tyrosine phosphorylation or the kinase activity of PKCdelta dramatically blocked PAR-mediated thromboxane A2 generation. We conclude that thrombin causes tyrosine phosphorylation of PKCdelta in a calcium- and Src-family kinase-dependent manner in platelets, with functional implications in thromboxane A2 generation. PMID- 15811958 TI - Indole-3-carbinol suppresses NF-kappaB and IkappaBalpha kinase activation, causing inhibition of expression of NF-kappaB-regulated antiapoptotic and metastatic gene products and enhancement of apoptosis in myeloid and leukemia cells. AB - Indole-3-carbinol, found in Brassica species vegetables (such as cabbage, cauliflower, and brussels spouts), exhibits antitumor effects through poorly defined mechanisms. Because several genes that regulate apoptosis, proliferation, and metastasis are regulated by nuclear factor-kappaB (NF-kappaB), we postulated that indole-3-carbinol must mediate its activity through NF-kappaB modulation. We demonstrated that indole-3-carbinol suppressed constitutive NF-kappaB activation and activation induced by tumor necrosis factor (TNF), interleukin-1beta (IL 1beta), phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS), and cigarette smoke; the suppression was not cell type specific, because activation was inhibited in myeloid, leukemia, and epithelial cells. This activation correlated with the sequential suppression of the IkappaBalpha kinase, IkappaBalpha phosphorylation, IkappaBalpha ubiquitination, IkappaBalpha degradation, p65 phosphorylation, p65 nuclear translocation, p65 acetylation, and NF-kappaB-dependent reporter gene expression. The NF-kappaB-regulated gene products cyclin D1, cyclooxygenase-2 (COX-2), matrix metalloproteinase-9 (MMP-9), survivin, inhibitor-of-apoptosis protein-1 (IAP1), IAP2, X chromosome-linked IAP (XIAP), Bcl-2, Bfl-1/A1, TNF receptor-associated factor-1 (TRAF1), and Fas associated death domain protein-like interleukin-1beta-converting enzyme inhibitory protein (FLIP) were all down-regulated by indole-3-carbinol. This down regulation led to the potentiation of apoptosis induced by cytokines and chemotherapeutic agents. Indole-3-carbinol suppressed constitutive NF-kappaB activation in mononuclear cells derived from bone marrow of acute myelogenous leukemia patients, and this correlated with inhibition of cell growth. Overall, our results indicated that indole-3-carbinol inhibits NF-kappaB and NF-kappaB regulated gene expression and that this mechanism may provide the molecular basis for its ability to suppress tumorigenesis. PMID- 15811959 TI - A phase-1 trial of bexarotene and denileukin diftitox in patients with relapsed or refractory cutaneous T-cell lymphoma. AB - Denileukin diftitox, a genetically engineered fusion protein combining the enzymatically active domains of diphtheria toxin and the full-length sequence for interleukin-2 (IL-2), efficiently targets lymphoma cells expressing the high affinity IL-2 receptor (IL-2R) consisting of the alpha/p55/CD25, beta/p75/CD122, and gamma/p64/CD132 chains. In vitro studies demonstrated that the retinoid X receptor (RXR) retinoid, bexarotene, at biologically relevant concentrations of 10(-6) M to 10(-8) M, upregulated both the p55 and p75 subunits of the IL-2R and enhanced 5- to 10-fold the susceptibility of T-cell leukemia cells to denileukin diftitox. To determine whether this biomodulatory effect could be recapitulated in vivo, we treated 14 patients with relapsed or refractory cutaneous T-cell lymphoma with escalating doses of bexarotene (75 mg/day-300 mg/day) and denileukin diftitox (18 mcg/kg per day x 3 days every 21 days) in a phase 1 trial. Overall response was 67% (4 complete responses, 4 partial responses). Modulation of IL-2R expression was observed at or above a bexarotene dose of 150 mg/day. Four patients experienced grade 2 or 3 leukopenia, and 2 had grade 4 lymphopenia. Our results demonstrate that the combination of denileukin diftitox and bexarotene is well tolerated and that even low doses (150 mg/day) of bexarotene are capable of in vivo upregulation of CD25 expression on circulating leukemia cells. PMID- 15811960 TI - Accelerated telomere shortening in glycosylphosphatidylinositol (GPI)-negative compared with GPI-positive granulocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) detected by proaerolysin flow-FISH. AB - Telomere length has been linked to disease stage and degree of (pan-)cytopenia in patients with bone marrow failure syndromes. The aim of the current study was to analyze the impact of replicative stress on telomere length in residual glycosylphosphatidylinositol-positive (GPI+) versus GPI- hematopoiesis in patients with paroxysmal nocturnal hemoglobinuria (PNH). Peripheral blood granulocytes from 16 patients and 22 healthy individuals were analyzed. For this purpose, we developed proaerolysin flow-FISH, a novel methodology that combines proaerolysin staining (for GPI expression) with flow-FISH (for telomere length measurement). We found significantly shortened telomeres in GPI- granulocytes (mean +/- SE: 6.26 +/- 0.27 telomere fluorescence units [TFU]), both compared with their GPI+ counterparts (6.88 +/- 0.38 TFU; P = .03) as well as with age matched healthy individuals (7.73 +/- 0.23 TFU; P < .001). Our findings are in support of a selective growth advantage model of PNH assuming that damage to the GPI+ hematopoietic stem-cell (HSC) compartment leads to compensatory hyperproliferation of residual GPI- HSCs. PMID- 15811962 TI - Haploinsufficiency of GATA-2 perturbs adult hematopoietic stem-cell homeostasis. AB - The zinc finger transcription factor GATA-2 plays a fundamental role in generating hematopoietic stem-cells in mammalian development. Less well defined is whether GATA-2 participates in adult stem-cell regulation, an issue we addressed using GATA-2 heterozygote mice that express reduced levels of GATA-2 in hematopoietic cells. While GATA-2+/- mice demonstrated decreases in some colony forming progenitors, the most prominent changes were observed within the stem cell compartment. Heterozygote bone marrow had a lower abundance of Lin(-)c kit(+)Sca-1(+)CD34- cells and performed poorly in competitive transplantation and quantitative week-5 cobblestone area-forming cell (CAFC) assays. Furthermore, a stem-cell-enriched population from GATA1+/- marrow was more quiescent and exhibited a greater frequency of apoptotic cells associated with decreased expression of the anti-apoptotic gene Bcl-xL. Yet the self-renewal potential of the +/- stem-cell compartment, as judged by serial transplantations, was unchanged. These data indicate compromised primitive cell proliferation and survival in the setting of a lower GATA-2 gene dose without a change in the differentiation or self-renewal capacity of the stem-cells that remain. Thus, GATA-2 dose regulates adult stem-cell homeostasis by affecting select aspects of stem cell function. PMID- 15811961 TI - Vav proteins are required for B-lymphocyte responses to LPS. AB - B lymphocytes respond to bacterial lipopolysaccharide (LPS) through Toll-like receptor 4 (TLR4) and CD180 (previously called RP105). We show here that the responses of B lymphocytes to LPS require the function of the Vav family of guanine nucleotide exchange factors. Vav1-mutant mice generate defective humoral immunoglobulin G (IgG) responses following administration of low doses of LPS but respond normally to higher doses, while mice lacking both Vav1 and Vav2 manifest defective responses even after a high dose of LPS. Vav1/2-mutant B cells fail to divide extensively in vitro in response to LPS or CD180, while deficiency of Vav1 alone impairs CD180-but not LPS-driven proliferation. Likewise, activation of Akt (a PI3K [phosphatidylinositol 3-kinase] target) and phosphorylation of IkappaBalpha in response to CD180 or LPS required Vav1 and Vav2, while Vav1 deficiency led to defective responses to CD180. In addition, activation of ERK (extracellular signal regulated kinase) required Vav1 and Vav2 in response to CD180 but was Vav1 and vav2 independent in response to LPS. Induction of CD86 and CD25 by anti-CD180 also required Vav function, as did the induction of the anti apoptotic protein Bcl-xL (B-cell leukemia XL). These data provide evidence for the function for the Vav proteins in regulating the responses of B cells to LPS. PMID- 15811964 TI - The cover. The Sick Child. PMID- 15811965 TI - FDA panel: keep COX-2 drugs on market: black box for COX-2 labels, caution urged for all NSAIDs. PMID- 15811966 TI - Pharmaceutical industry funding for residencies sparks controversy. PMID- 15811967 TI - Happy 50th birthday, Salk vaccine. PMID- 15811971 TI - Cardiovascular outcomes and antihypertensive drug treatment in older women. PMID- 15811972 TI - Cardiovascular outcomes and antihypertensive drug treatment in older women. PMID- 15811973 TI - Comparison of diets for weight loss and heart disease risk reduction. PMID- 15811974 TI - Comparison of diets for weight loss and heart disease risk reduction. PMID- 15811975 TI - Resurrecting treatment histories of dead patients. PMID- 15811976 TI - Resurrecting treatment histories of dead patients. PMID- 15811977 TI - Postponement of cancer death and life insurance. PMID- 15811978 TI - Readability of notice of privacy forms used by major health care institutions. PMID- 15811979 TI - Outcomes in hypertensive black and nonblack patients treated with chlorthalidone, amlodipine, and lisinopril. AB - CONTEXT: Few cardiovascular outcome data are available for blacks with hypertension treated with angiotensin-converting enzyme (ACE) inhibitors or calcium channel blockers (CCBs). OBJECTIVE: To determine whether an ACE inhibitor or CCB is superior to a thiazide-type diuretic in reducing cardiovascular disease (CVD) incidence in racial subgroups. DESIGN, SETTING, AND PARTICIPANTS: Prespecified subgroup analysis of ALLHAT, a randomized, double-blind, active controlled, clinical outcome trial conducted between February 1994 and March 2002 in 33,357 hypertensive US and Canadian patients aged 55 years or older (35% black) with at least 1 other cardiovascular risk factor. INTERVENTIONS: Antihypertensive regimens initiated with a CCB (amlodipine) or an ACE inhibitor (lisinopril) vs a thiazide-type diuretic (chlorthalidone). Other medications were added to achieve goal blood pressures (BPs) less than 140/90 mm Hg. MAIN OUTCOME MEASURES: The primary outcome was combined fatal coronary heart disease (CHD) or nonfatal myocardial infarction (MI), analyzed by intention-to-treat. Secondary outcomes included all-cause mortality, stroke, combined CVD (CHD death, nonfatal MI, stroke, angina, coronary revascularization, heart failure [HF], or peripheral vascular disease), and end-stage renal disease. RESULTS: No significant difference was found between treatment groups for the primary CHD outcome in either racial subgroup. For amlodipine vs chlorthalidone only, HF was the only prespecified clinical outcome that differed significantly (overall: relative risk [RR], 1.37; 95% confidence interval [CI], 1.24-1.51; blacks: RR, 1.46; 95% CI, 1.24-1.73; nonblacks: RR, 1.32; 95% CI, 1.17-1.49; P<.001 for each comparison) with no difference in treatment effects by race (P = .38 for interaction). For lisinopril vs chlorthalidone, results differed by race for systolic BP (greater decrease in blacks with chlorthalidone), stroke, and combined CVD outcomes (P<.001, P = .01, and P = .04, respectively, for interactions). In blacks and nonblacks, respectively, the RRs for stroke were 1.40 (95% CI, 1.17-1.68) and 1.00 (95% CI, 0.85-1.17) and for combined CVD were 1.19 (95% CI, 1.09-1.30) and 1.06 (95% CI, 1.00-1.13). For HF, the RRs were 1.30 (95% CI, 1.10-1.54) and 1.13 (95% CI, 1.00-1.28), with no significant interaction by race. Time-dependent BP adjustment did not significantly alter differences in outcome for lisinopril vs chlorthalidone in blacks. CONCLUSIONS: In blacks and nonblack subgroups, rates were not lower in the amlodipine or lisinopril groups than in the chlorthalidone group for either the primary CHD or any other prespecified clinical outcome, and diuretic-based treatment resulted in the lowest risk of heart failure. While the improved outcomes with chlorthalidone were more pronounced for some outcomes in blacks than in nonblacks, thiazide-type diuretics remain the drugs of choice for initial therapy of hypertension in both black and nonblack hypertensive patients. PMID- 15811980 TI - N-terminal pro-brain natriuretic peptide, C-reactive protein, and urinary albumin levels as predictors of mortality and cardiovascular events in older adults. AB - CONTEXT: B-type natriuretic peptides have been shown to predict cardiovascular disease in apparently healthy individuals but their predictive ability for mortality and future cardiovascular events compared with C-reactive protein (CRP) and urinary albumin/creatinine ratio is unknown. OBJECTIVE: To assess the prognostic value of the N-amino terminal fragment of the prohormone brain natriuretic peptide (NT-proBNP) vs CRP and urinary albumin/creatinine ratio in an older adult population. DESIGN, SETTING, AND PARTICIPANTS: A population-based prospective study of 764 participants aged 50 to 89 years from a community in Copenhagen, Denmark, in which 658 participants provided blood and urinary samples and were examined between September 1, 1998, and January 24, 2000. Of these participants, 626 without heart or renal failure were enrolled. A subgroup of 537 had no history of cardiovascular disease at baseline. During 5 years of follow-up (to December 31, 2003), 94 participants died and 65 developed a first major cardiovascular event. MAIN OUTCOME MEASURES: Risk of mortality and first major cardiovascular event by baseline levels of NT-proBNP, CRP, and urinary albumin/creatinine ratio levels. RESULTS: After adjustment for the cardiovascular risk factors of age, sex, smoking, diabetes mellitus, hypertension or ischemic heart disease, total cholesterol, and serum creatinine, the hazard ratio (HR) of mortality for values above the 80th percentile of NT-proBNP was 1.96 (95% confidence interval [CI], 1.21-3.19); for CRP, 1.46 (95% CI, 0.89-2.24); and for urinary albumin/creatinine ratio, 1.88 (95% CI, 1.18-2.98). Additional adjustment for left ventricular systolic dysfunction did not markedly attenuate the predictive value of NT-proBNP (HR, 1.82; 95% CI, 1.11-2.98). The absolute unadjusted increase in mortality risk for participants with values above the 80th percentile vs equal to or below the 80th percentile was 24.5% for NT-proBNP, 7.8% for CRP, and 19.5% for urinary albumin/creatinine ratio. The NT-proBNP levels were associated with first major cardiovascular events (nonfatal myocardial infarction, fatal coronary heart disease, unstable angina, heart failure, stroke, and transient ischemic attack) with an adjusted HR of 3.24 (95% CI, 1.80-5.79) vs 1.02 (95% CI, 0.56-1.85) for CRP and 2.32 (95% CI, 1.33-4.05) for urinary albumin/creatinine ratio when comparing participants with values above the 80th percentile with those with values equal to or below the 80th percentile. CONCLUSIONS: Measurements of NT-proBNP provide prognostic information of mortality and first major cardiovascular events beyond traditional risk factors. NT-proBNP was a stronger risk biomarker for cardiovascular disease and death than CRP was in nonhospitalized individuals aged 50 to 89 years. PMID- 15811981 TI - Efficacy and tolerability of long-acting injectable naltrexone for alcohol dependence: a randomized controlled trial. AB - CONTEXT: Alcohol dependence is a common disorder associated with significant morbidity and mortality. Naltrexone, an opioid antagonist, has been shown to be effective for treatment of alcohol dependence. However, adherence to daily oral pharmacotherapy can be problematic, and clinical acceptance and utility of oral naltrexone have been limited. OBJECTIVE: To determine efficacy and tolerability of a long-acting intramuscular formulation of naltrexone for treatment of alcohol dependent patients. DESIGN, SETTING, AND PARTICIPANTS: A 6-month, randomized, double-blind, placebo-controlled trial conducted between February 2002 and September 2003 at 24 US public hospitals, private and Veterans Administration clinics, and tertiary care medical centers. Of the 899 individuals screened, 627 who were diagnosed as being actively drinking alcohol-dependent adults were randomized to receive treatment and 624 received at least 1 injection. INTERVENTION: An intramuscular injection of 380 mg of long-acting naltrexone (n = 205) or 190 mg of long-acting naltrexone (n = 210) or a matching volume of placebo (n = 209) each administered monthly and combined with 12 sessions of low intensity psychosocial intervention. MAIN OUTCOME MEASURE: The event rate of heavy drinking days in the intent-to-treat population. RESULTS: Compared with placebo, 380 mg of long-acting naltrexone resulted in a 25% decrease in the event rate of heavy drinking days (P = .02) [corrected] and 190 mg of naltrexone resulted in a 17% decrease (P = .07). Sex and pretreatment abstinence each showed significant interaction with the medication group on treatment outcome, with men and those with lead-in abstinence both exhibiting greater treatment effects. Discontinuation due to adverse events occurred in 14.1% in the 380-mg and 6.7% in the 190-mg group and 6.7% in the placebo group. Overall, rate and time to treatment discontinuation were similar among treatment groups. CONCLUSIONS: Long acting naltrexone was well tolerated and resulted in reductions in heavy drinking among treatment-seeking alcohol-dependent patients during 6 months of therapy. These data indicate that long-acting naltrexone can be of benefit in the treatment of alcohol dependence. PMID- 15811982 TI - Effects of exercise and stress management training on markers of cardiovascular risk in patients with ischemic heart disease: a randomized controlled trial. AB - CONTEXT: Observational studies have shown that psychosocial factors are associated with increased risk for cardiovascular morbidity and mortality, but the effects of behavioral interventions on psychosocial and medical end points remain uncertain. OBJECTIVE: To determine the effect of 2 behavioral programs, aerobic exercise training and stress management training, with routine medical care on psychosocial functioning and markers of cardiovascular risk. DESIGN, SETTING, AND PATIENTS: Randomized controlled trial of 134 patients (92 male and 42 female; aged 40-84 years) with stable ischemic heart disease (IHD) and exercise-induced myocardial ischemia. Conducted from January 1999 to February 2003. INTERVENTIONS: Routine medical care (usual care); usual care plus supervised aerobic exercise training for 35 minutes 3 times per week for 16 weeks; usual care plus weekly 1.5-hour stress management training for 16 weeks. MAIN OUTCOME MEASURES: Self-reported measures of general distress (General Health Questionnaire [GHQ]) and depression (Beck Depression Inventory [BDI]); left ventricular ejection fraction (LVEF) and wall motion abnormalities (WMA); flow mediated dilation; and cardiac autonomic control (heart rate variability during deep breathing and baroreflex sensitivity). RESULTS: Patients in the exercise and stress management groups had lower mean (SE) BDI scores (exercise: 8.2 [0.6]; stress management: 8.2 [0.6]) vs usual care (10.1 [0.6]; P = .02); reduced distress by GHQ scores (exercise: 56.3 [0.9]; stress management: 56.8 [0.9]) vs usual care (53.6 [0.9]; P = .02); and smaller reductions in LVEF during mental stress testing (exercise: -0.54% [0.44%]; stress management: -0.34% [0.45%]) vs usual care (-1.69% [0.46%]; P = .03). Exercise and stress management were associated with lower mean (SE) WMA rating scores (exercise: 0.20 [0.07]; stress management: 0.10 [0.07]) in a subset of patients with significant stress-induced WMA at baseline vs usual care (0.36 [0.07]; P = .02). Patients in the exercise and stress management groups had greater mean (SE) improvements in flow-mediated dilation (exercise: mean [SD], 5.6% [0.45%]; stress management: 5.2% [0.47%]) vs usual care patients (4.1% [0.48%]; P = .03). In a subgroup, those receiving stress management showed improved mean (SE) baroreflex sensitivity (8.2 [0.8] ms/mm Hg) vs usual care (5.1 [0.9] ms/mm Hg; P = .02) and significant increases in heart rate variability (193.7 [19.6] ms) vs usual care (132.1 [21.5] ms; P = .04). CONCLUSION: For patients with stable IHD, exercise and stress management training reduced emotional distress and improved markers of cardiovascular risk more than usual medical care alone. PMID- 15811983 TI - Evaluating iatrogenic risk of youth suicide screening programs: a randomized controlled trial. AB - CONTEXT: Universal screening for mental health problems and suicide risk is at the forefront of the national agenda for youth suicide prevention, yet no study has directly addressed the potential harm of suicide screening. OBJECTIVE: To examine whether asking about suicidal ideation or behavior during a screening program creates distress or increases suicidal ideation among high school students generally or among high-risk students reporting depressive symptoms, substance use problems, or suicide attempts. DESIGN, SETTING, AND PARTICIPANTS: A randomized controlled study conducted within the context of a 2-day screening strategy. Participants were 2342 students in 6 high schools in New York State in 2002-2004. Classes were randomized to an experimental group (n = 1172), which received the first survey with suicide questions, or to a control group (n = 1170), which did not receive suicide questions. MAIN OUTCOME MEASURES: Distress measured at the end of the first survey and at the beginning of the second survey 2 days after the first measured on the Profile of Mood States adolescent version (POMS-A) instrument. Suicidal ideation assessed in the second survey. RESULTS: Experimental and control groups did not differ on distress levels immediately after the first survey (mean [SD] POMS-A score, 5.5 [9.7] in the experimental group and 5.1 [10.0] in the control group; P = .66) or 2 days later (mean [SD] POMS-A score, 4.3 [9.0] in the experimental group and 3.9 [9.4] in the control group; P = .41), nor did rates of depressive feelings differ (13.3% and 11.0%, respectively; P = .19). Students exposed to suicide questions were no more likely to report suicidal ideation after the survey than unexposed students (4.7% and 3.9%, respectively; P = .49). High-risk students (defined as those with depression symptoms, substance use problems, or any previous suicide attempt) in the experimental group were neither more suicidal nor distressed than high-risk youth in the control group; on the contrary, depressed students and previous suicide attempters in the experimental group appeared less distressed (P = .01) and suicidal (P = .02), respectively, than high-risk control students. CONCLUSIONS: No evidence of iatrogenic effects of suicide screening emerged. Screening in high schools is a safe component of youth suicide prevention efforts. PMID- 15811984 TI - Association between compensation status and outcome after surgery: a meta analysis. AB - CONTEXT: Compensation, whether through workers' compensation or through litigation, has been associated with poor outcome after surgery; however, this association has not been examined by meta-analysis. OBJECTIVE: To investigate the association between compensation status and outcome after surgery. DATA SOURCES: We searched MEDLINE (1966-2003), EMBASE (1980-2003), CINAHL, the Cochrane Controlled Trials Register, and reference lists of retrieved articles and textbooks, and we contacted experts in the field. STUDY SELECTION: The review included any trial of surgical intervention in which compensation status was reported and results were compared according to that status. No restrictions were placed on study design, language, or publication date. Studies were selected by 2 unblinded independent reviewers. DATA EXTRACTION: Two reviewers independently extracted data on study type, study quality, surgical procedure, outcome, country of origin, length and completeness of follow-up, and compensation type. DATA SYNTHESIS: Two hundred eleven studies satisfied the inclusion criteria. Of these, 175 stated that the presence of compensation (workers' compensation with or without litigation) was associated with a worse outcome, 35 found no difference or did not describe a difference, and 1 described a benefit associated with compensation. A meta-analysis of 129 studies with available data (n = 20,498 patients) revealed the summary odds ratio for an unsatisfactory outcome in compensated patients to be 3.79 (95% confidence interval, 3.28-4.37 by random effects model). Grouping studies by country, procedure, length of follow-up, completeness of follow-up, study type, and type of compensation showed the association to be consistent for all subgroups. CONCLUSIONS: Compensation status is associated with poor outcome after surgery. This effect is significant, clinically important, and consistent. Because data were obtained from observational studies and were not homogeneous, the summary effect should be interpreted with caution. Compensation status should be considered a potential confounder in all studies of surgical intervention. Determination of the mechanism for this association requires further study. PMID- 15811985 TI - The clinical sequelae of intravascular hemolysis and extracellular plasma hemoglobin: a novel mechanism of human disease. AB - CONTEXT: The efficient sequestration of hemoglobin by the red blood cell membrane and the presence of multiple hemoglobin clearance mechanisms suggest a critical need to prevent the buildup of this molecule in the plasma. A growing list of clinical manifestations attributed to hemoglobin release in a variety of acquired and iatrogenic hemolytic disorders suggests that hemolysis and hemoglobinemia should be considered as a novel mechanism of human disease. EVIDENCE ACQUISITION: Pertinent scientific literature databases and references were searched through October 2004 using terms that encompassed various aspects of hemolysis, hemoglobin preparations, clinical symptoms associated with plasma hemoglobin, nitric oxide in hemolysis, anemia, pulmonary hypertension, paroxysmal nocturnal hemoglobinuria, and sickle-cell disease. EVIDENCE SYNTHESIS: Hemoglobin is released into the plasma from the erythrocyte during intravascular hemolysis in hereditary, acquired, and iatrogenic hemolytic conditions. When the capacity of protective hemoglobin-scavenging mechanisms has been saturated, levels of cell free hemoglobin increase in the plasma, resulting in the consumption of nitric oxide and clinical sequelae. Nitric oxide plays a major role in vascular homeostasis and has been shown to be a critical regulator of basal and stress mediated smooth muscle relaxation and vasomotor tone, endothelial adhesion molecule expression, and platelet activation and aggregation. Thus, clinical consequences of excessive cell-free plasma hemoglobin levels during intravascular hemolysis or the administration of hemoglobin preparations include dystonias involving the gastrointestinal, cardiovascular, pulmonary, and urogenital systems, as well as clotting disorders. Many of the clinical sequelae of intravascular hemolysis in a prototypic hemolytic disease, paroxysmal nocturnal hemoglobinuria, are readily explained by hemoglobin-mediated nitric oxide scavenging. CONCLUSION: A growing body of evidence supports the existence of a novel mechanism of human disease, namely, hemolysis-associated smooth muscle dystonia, vasculopathy, and endothelial dysfunction. PMID- 15811986 TI - Diuretics are color blind. PMID- 15811987 TI - Cardiovascular risk stratification in older patients: role of brain natriuretic peptide, C-reactive protein, and urinary albumin levels. PMID- 15811988 TI - JAMA patient page. Alcohol abuse and alcoholism. PMID- 15811989 TI - Biomarkers for in situ detection of anaerobic ammonium-oxidizing (anammox) bacteria. PMID- 15811990 TI - Endophytic colonization of Vitis vinifera L. by plant growth-promoting bacterium Burkholderia sp. strain PsJN. AB - Patterns of colonization of Vitis vinifera L. cv. Chardonnay plantlets by a plant growth-promoting bacterium, Burkholderia sp. strain PsJN, were studied under gnotobiotic conditions. Wild-type strain PsJN and genetically engineered derivatives of this strain tagged with gfp (PsJN::gfp2x) or gusA (PsJN::gusA11) genes were used to enumerate and visualize tissue colonization. The rhizospheres of 4- to 5-week-old plantlets with five developed leaves were inoculated with bacterial suspensions. Epiphytic and endophytic colonization patterns were then monitored by dilution plating assays and microscopic observation of organ sections. Bacteria were chronologically detected first on root surfaces, then in root internal tissues, and finally in the fifth internode and the tissues of the fifth leaf. Analysis of the PsJN colonization patterns showed that this strain colonizes grapevine root surfaces, as well as cell walls and the whole surface of some rhizodermal cells. Cells were also abundant at lateral root emergence sites and root tips. Furthermore, cell wall-degrading endoglucanase and endopolygalacturonase secreted by PsJN explained how the bacterium gains entry into root internal tissues. Host defense reactions were observed in the exodermis and in several cortical cell layers. Bacteria were not observed on stem and leaf surfaces but were found in xylem vessels of the fifth internode and the fifth leaf of plantlets. Moreover, bacteria were more abundant in the fifth leaf than in the fifth internode and were found in substomatal chambers. Thus, it seems that Burkholderia sp. strain PsJN induces a local host defense reaction and systemically spreads to aerial parts through the transpiration stream. PMID- 15811991 TI - Dual symbiosis in a Bathymodiolus sp. mussel from a methane seep on the Gabon continental margin (Southeast Atlantic): 16S rRNA phylogeny and distribution of the symbionts in gills. AB - Deep-sea mussels of the genus Bathymodiolus (Bivalvia: Mytilidae) harbor symbiotic bacteria in their gills and are among the dominant invertebrate species at cold seeps and hydrothermal vents. An undescribed Bathymodiolus species was collected at a depth of 3,150 m in a newly discovered cold seep area on the southeast Atlantic margin, close to the Zaire channel. Transmission electron microscopy, comparative 16S rRNA analysis, and fluorescence in situ hybridization indicated that this Bathymodiolus sp. lives in a dual symbiosis with sulfide- and methane-oxidizing bacteria. A distinct distribution pattern of the symbiotic bacteria in the gill epithelium was observed, with the thiotrophic symbiont dominating the apical region and the methanotrophic symbiont more abundant in the basal region of the bacteriocytes. No variations in this distribution pattern or in the relative abundances of the two symbionts were observed in mussels collected from three different mussel beds with methane concentrations ranging from 0.7 to 33.7 microM. The 16S rRNA sequence of the methanotrophic symbiont is most closely related to those of known methanotrophic symbionts from other bathymodiolid mussels. Surprisingly, the thiotrophic Bathymodiolus sp. 16S rRNA sequence does not fall into the monophyletic group of sequences from thiotrophic symbionts of all other Bathymodiolus hosts. While these mussel species all come from vents, this study describes the first thiotrophic sequence from a seep mussel and shows that it is most closely related (99% sequence identity) to an environmental clone sequence obtained from a hydrothermal plume near Japan. PMID- 15811992 TI - Putative polyketide synthase and laccase genes for biosynthesis of aurofusarin in Gibberella zeae. AB - Mycelia of Gibberella zeae (anamorph, Fusarium graminearum), an important pathogen of cereal crops, are yellow to tan with white to carmine red margins. We isolated genes encoding the following two proteins that are required for aurofusarin biosynthesis from G. zeae: a type I polyketide synthase (PKS) and a putative laccase. Screening of insertional mutants of G. zeae, which were generated by using a restriction enzyme-mediated integration procedure, resulted in the isolation of mutant S4B3076, which is a pigment mutant. In a sexual cross of the mutant with a strain with normal pigmentation, the pigment mutation was linked to the inserted vector. The vector insertion site in S4B3076 was a HindIII site 38 bp upstream from an open reading frame (ORF) on contig 1.116 in the F. graminearum genome database. The ORF, designated Gip1 (for Gibberella zeae pigment mutation 1), encodes a putative laccase. A 30-kb region surrounding the insertion site and Gip1 contains 10 additional ORFs, including a putative ORF identified as PKS12 whose product exhibits about 40% amino acid identity to the products of type I fungal PKS genes, which are involved in pigment biosynthesis. Targeted gene deletion and complementation analyses confirmed that both Gip1 and PKS12 are required for aurofusarin production in G. zeae. This information is the first information concerning the biosynthesis of these pigments by G. zeae and could help in studies of their toxicity in domesticated animals. PMID- 15811993 TI - Incorporation of glucose under anoxic conditions by bacterioplankton from coastal North Sea surface waters. AB - It has been hypothesized that the potential for anaerobic metabolism might be a common feature of bacteria in coastal marine waters (L. Riemann and F. Azam, Appl. Environ. Microbiol. 68: 5554-5562, 2002). Therefore, we investigated whether different phylogenetic groups of heterotrophic picoplankton from the coastal North Sea were able to take up a simple carbon source under anoxic conditions. Oxic and anoxic incubations (4 h) or enrichments (24 h) of seawater with radiolabeled glucose were performed in July and August 2003. Bacteria with incorporated substrate were identified by using a novel protocol in which we combined fluorescence in situ hybridization and microautoradiography of cells on membrane filters. Incorporation of glucose under oxic and anoxic conditions was found in alpha-Proteobacteria, gamma-Proteobacteria, and the Cytophaga Flavobacterium cluster of the Bacteroidetes at both times, but not in marine Euryarchaeota. In July, the majority of cells belonging to the alpha proteobacterial Roseobacter clade showed tracer incorporation both in oxic incubations and in oxic and anoxic enrichments. In August, only a minority of the Roseobacter cells, but most bacteria affiliated with Vibrio spp., were able to incorporate the tracer under either condition. A preference for glucose uptake under anoxic conditions was observed for bacteria related to Alteromonas and the Pseudoalteromonas-Colwellia group. These genera are commonly considered to be strictly aerobic, but facultatively fermentative strains have been described. Our findings suggest that the ability to incorporate substrates anaerobically is widespread in pelagic marine bacteria belonging to different phylogenetic groups. Such bacteria may be abundant in fully aerated coastal marine surface waters. PMID- 15811994 TI - Proteomic profiling of recombinant Escherichia coli in high-cell-density fermentations for improved production of an antibody fragment biopharmaceutical. AB - By using two-dimensional polyacrylamide gel electrophoresis, a proteomic analysis over time was conducted with high-cell-density, industrial, phosphate-limited Escherichia coli fermentations at the 10-liter scale. During production, a recombinant, humanized antibody fragment was secreted and assembled in a soluble form in the periplasm. E. coli protein changes associated with culture conditions were distinguished from protein changes associated with heterologous protein expression. Protein spots were monitored quantitatively and qualitatively. Differentially expressed proteins were quantitatively assessed by using a t-test method with a 1% false discovery rate as a significance criterion. As determined by this criterion, 81 protein spots changed significantly between 14 and 72 h (final time) of the control fermentations (vector only). Qualitative (on-off) comparisons indicated that 20 more protein spots were present only at 14 or 72 h in the control fermentations. These changes reflected physiological responses to the culture conditions. In control and production fermentations at 72 h, 25 protein spots were significantly differentially expressed. In addition, 19 protein spots were present only in control or production fermentations at this time. The quantitative and qualitative changes were attributable to overexpression of recombinant protein. The physiological changes observed during the fermentations included the up-regulation of phosphate starvation proteins and the down-regulation of ribosomal proteins and nucleotide biosynthesis proteins. Synthesis of the stress protein phage shock protein A (PspA) was strongly correlated with synthesis of a recombinant product. This suggested that manipulation of PspA levels might improve the soluble recombinant protein yield in the periplasm for this bioprocess. Indeed, controlled coexpression of PspA during production led to a moderate, but statistically significant, improvement in the yield. PMID- 15811995 TI - Competitive interactions in mixed-species biofilms containing the marine bacterium Pseudoalteromonas tunicata. AB - Pseudoalteromonas tunicata is a biofilm-forming marine bacterium that is often found in association with the surface of eukaryotic organisms. It produces a range of extracellular inhibitory compounds, including an antibacterial protein (AlpP) thought to be beneficial for P. tunicata during competition for space and nutrients on surfaces. As part of our studies on the interactions between P. tunicata and the epiphytic bacterial community on the marine plant Ulva lactuca, we investigated the hypothesis that P. tunicata is a superior competitor compared with other bacteria isolated from the plant. A number of U. lactuca bacterial isolates were (i) identified by 16S rRNA gene sequencing, (ii) characterized for the production of or sensitivity to extracellular antibacterial proteins, and (iii) labeled with a fluorescent color tag (either the red fluorescent protein DsRed or green fluorescent protein). We then grew single- and mixed-species bacterial biofilms containing P. tunicata in glass flow cell reactors. In pure culture, all the marine isolates formed biofilms containing microcolony structures within 72 h. However, in mixed-species biofilms, P. tunicata removed the competing strain unless its competitor was relatively insensitive to AlpP (Pseudoalteromonas gracilis) or produced strong inhibitory activity against P. tunicata (Roseobacter gallaeciensis). Moreover, biofilm studies conducted with an AlpP- mutant of P. tunicata indicated that the mutant was less competitive when it was introduced into preestablished biofilms, suggesting that AlpP has a role during competitive biofilm formation. When single-species biofilms were allowed to form microcolonies before the introduction of a competitor, these microcolonies coexisted with P. tunicata for extended periods of time before they were removed. Two marine bacteria (R. gallaeciensis and P. tunicata) were superior competitors in this study. Our data suggest that this dominance can be attributed to the ability of these organisms to rapidly form microcolonies and their ability to produce extracellular antibacterial compounds. PMID- 15811996 TI - Biocatalytic production of perillyl alcohol from limonene by using a novel Mycobacterium sp. cytochrome P450 alkane hydroxylase expressed in Pseudomonas putida. AB - A number of oxygenated monoterpenes present at low concentrations in plant oils have anticarcinogenic properties. One of the most promising compounds in this respect is (-)-perillyl alcohol. Since this natural product is present only at low levels in a few plant oils, an alternative, synthetic source is desirable. Screening of 1,800 bacterial strains showed that many alkane degraders were able to specifically hydroxylate l-limonene in the 7 position to produce enantiopure ( )-perillyl alcohol. The oxygenase responsible for this was purified from the best performing wild-type strain, Mycobacterium sp. strain HXN-1500. By using N terminal sequence information, a 6.2-kb ApaI fragment was cloned, which encoded a cytochrome P450, a ferredoxin, and a ferredoxin reductase. The three genes were successfully coexpressed in Pseudomonas putida by using the broad-host-range vector pCom8, and the recombinant converted limonene to perillyl alcohol with a specific activity of 3 U/g (dry weight) of cells. The construct was subsequently used in a 2-liter bioreactor to produce perillyl alcohol on a scale of several grams. PMID- 15811997 TI - Optimization of cholesterol removal by probiotics in the presence of prebiotics by using a response surface method. AB - Lactobacillus casei ASCC 292 was grown in the presence of six prebiotics, namely, sorbitol, mannitol, maltodextrin, high-amylose maize, fructooligosaccharide (FOS), and inulin, in order to determine the combination of probiotic and prebiotics that would remove the highest level of cholesterol. A first-order model showed that the combination of L. casei ASCC 292, FOS, and maltodextrin was the most efficient for the removal of cholesterol, and the optimum experimental region was developed by using the steepest ascent. This led to the middle points of probiotic (1.70% [wt/vol]), FOS (4.80% [wt/vol]), and maltodextrin (6.80% [wt/vol]) for the development of a central composite design for optimization. Perturbation plot, response surface, and coefficient estimates showed that all three factors had significant quadratic effects on cholesterol removal, with FOS showing the most conspicuous quadratic change. A second-order polynomial regression model estimated that the optimum condition of the factors for cholesterol removal by L. casei ASCC 292 is 1.71% (wt/vol) probiotic, 4.95% (wt/vol) FOS, and 6.62% (wt/vol) maltodextrin. Validation experiments showed that the predicted optimum conditions were more efficient than the high and low levels of the factors and the center points. A response surface method proved reliable for developing the model, optimizing factors, and analyzing interaction effects. Analyses of growth, substrate utilization, growth yield, mean doubling time, and short-chain fatty acid (SCFA) production by the use of quadratic models indicated that cholesterol removal was growth associated. The concentration of L. casei ASCC 292 had the most significant quadratic effect on all responses studied, except for substrate utilization and SCFA production, which were significantly (P < 0.05) influenced by the interactions between the probiotic and both prebiotics, indicating that they were closely associated with the uptake of prebiotics. PMID- 15811998 TI - Web-type evolution of rhodococcus gene clusters associated with utilization of naphthalene. AB - Clusters of genes which include determinants for the catalytic subunits of naphthalene dioxygenase (narAa and narAb) were analyzed in naphthalene-degrading Rhodococcus strains. We demonstrated (i) that in the region analyzed homologous gene clusters are separated from each other by nonhomologous DNA, (ii) that there are various degrees of homology between related genes, and (iii) that nar genes are located on plasmids in strains NCIMB12038 and P400 and on a chromosome in P200. These observations suggest that genetic exchange and reshuffling of genetic modules, as well as vertical descent of the genetic information, were the main routes in the evolution of naphthalene degradation in Rhodococcus. These conclusions were supported by studies of transcription patterns in the region analyzed. It was found that the nar region is not organized into a single operon but there are several transcription units which differ in the strains investigated. The narA and narB genes were found to be transcribed as a single unit in all strains analyzed, and their transcription was induced by naphthalene. The putative aldolase gene (narC) was found on the same transcript only in strains P200 and P400. In NCIMB12038 transcription of two more gene clusters was induced by growth on naphthalene. Transcription start sites for narA and narB were found to be different in all of the strains studied. Putative regulatory genes (narR1 and narR2) were transcribed as a single mRNA in naphthalene-induced cells. At the same time, a number of the genes known to be essential for naphthalene catabolism in gram-negative bacteria were not found in the region analyzed. PMID- 15811999 TI - Characterization of Cry34/Cry35 binary insecticidal proteins from diverse Bacillus thuringiensis strain collections. AB - Bacillus thuringiensis crystal proteins of the Cry34 and Cry35 classes function as binary toxins showing activity on the western corn rootworm, Diabrotica virgifera virgifera LeConte. We surveyed 6,499 B. thuringiensis isolates by hybridization for sequences related to cry35A genes, identifying 78 strains. Proteins of the appropriate molecular mass (ca. 44 kDa) for Cry35 were observed in 42 of the strains. Full-length, or nearly full-length, sequences of 34 cry34 genes and 16 cry35 genes were also obtained from cloning, PCR analysis, and DNA sequencing. These included representatives of all known Cry34A, Cry34B, Cry35A, and Cry35B classes, as well as a novel Cry34A/Cry35A-like pair. Bioassay analysis indicated that cry35-hybridizing strains not producing a ca. 14-kDa protein, indicative of Cry34, were not active on corn rootworms, and that the previously identified Cry34A/Cry35A pairs were more active than the Cry34B/Cry35B pairs. The cry35-hybridizing B. thuringiensis strains were found in locales and materials typical for other B. thuringiensis strains. Comparison of the sequences with the geographic origins of the strains showed that identical, or nearly identical, sequences were found in strains from both Australasia and the Americas. Sequence similarity searches revealed that Cry34 proteins are similar to predicted proteins in Photorhabdus luminescens and Dictyostelium discoidium, and that Cry35Ab1 contains a segment similar to beta-trefoil domains that may be a binding motif. The binary Cry34/Cry35 B. thuringiensis crystal proteins thus appear closely related to each other, are environmentally ubiquitous, and share sequence similarities consistent with activity through membrane disruption in target organisms. PMID- 15812000 TI - Lignin-derived compounds as efficient laccase mediators for decolorization of different types of recalcitrant dyes. AB - Ten phenols were selected as natural laccase mediators after screening 44 different compounds with a recalcitrant dye (Reactive Black 5) as a substrate. Their performances were evaluated at different mediator/dye ratios and incubation times (up to 6 h) by the use of Pycnoporus cinnabarinus and Trametes villosa laccases and were compared with those of eight known synthetic mediators (including -NOH- compounds). Among the six types of dyes assayed, only Reactive Blue 38 (phthalocyanine) was resistant to laccase-mediator treatment under the conditions used. Acid Blue 74 (indigoid dye), Reactive Blue 19 (anthraquinoid dye), and Aniline Blue (triarylmethane-type dye) were partially decolorized by the laccases alone, although decolorization was much more efficient and rapid with mediators, whereas Reactive Black 5 (diazo dye) and Azure B (heterocyclic dye) could be decolorized only in the presence of mediators. The efficiency of each natural mediator depended on the type of dye to be treated but, with the only exception being Azure B (< 50% decolorization), nearly complete decolorization (80 to 100%) was attained in all cases. Similar rates were attained with the best synthetic mediators, but the reactions were significantly slower. Phenolic aldehydes, ketones, acids, and esters related to the three lignin units were among the best mediators, including p-coumaric acid, vanillin, acetovanillone, methyl vanillate, and above all, syringaldehyde and acetosyringone. The last two compounds are especially promising as ecofriendly (and potentially cheap) mediators for industrial applications since they provided the highest decolorization rates in only 5 to 30 min, depending on the type of dye to be treated. PMID- 15812001 TI - Heat inactivation of Mycobacterium avium subsp. paratuberculosis in milk. AB - The effectiveness of pasteurization and the concentration of Mycobacterium avium subsp. paratuberculosis in raw milk have been identified in quantitative risk analysis as the most critical factors influencing the potential presence of viable Mycobacterium paratuberculosis in dairy products. A quantitative assessment of the lethality of pasteurization was undertaken using an industrial pasteurizer designed for research purposes with a validated Reynolds number of 62,112 and flow rates of 3,000 liters/h. M. paratuberculosis was artificially added to raw whole milk, which was then homogenized, pasteurized, and cultured, using a sensitive technique capable of detecting one organism per 10 ml of milk. Twenty batches of milk containing 10(3) to 10(4) organisms/ml were processed with combinations of three temperatures of 72, 75, and 78 degrees C and three time intervals of 15, 20, and 25 s. Thirty 50-ml milk samples from each processed batch were cultured, and the logarithmic reduction in M. paratuberculosis organisms was determined. In 17 of the 20 runs, no viable M. paratuberculosis organisms were detected, which represented > 6-log10 reductions during pasteurization. These experiments were conducted with very heavily artificially contaminated milk to facilitate the measurement of the logarithmic reduction. In three of the 20 runs of milk, pasteurized at 72 degrees C for 15 s, 75 degrees C for 25 s, and 78 degrees C for 15 s, a few viable organisms (0.002 to 0.004 CFU/ml) were detected. Pasteurization at all temperatures and holding times was found to be very effective in killing M. paratuberculosis, resulting in a reduction of > 6 log10 in 85% of runs and > 4 log10 in 14% of runs. PMID- 15812002 TI - Direct microbial reduction and subsequent preservation of uranium in natural near surface sediment. AB - The fate of uranium in natural systems is of great environmental importance. X ray absorption near-edge spectroscopy (XANES) revealed that U(VI) was reduced to U(IV) in shallow freshwater sediment at an open pit in an inactive uranium mine. Geochemical characterization of the sediment showed that nitrate, Fe(III), and sulfate had also been reduced in the sediment. Observations of the sediment particles and microbial cells by scanning and transmission electron microscopy, coupled with elemental analysis by energy dispersive spectroscopy, revealed that uranium was concentrated at microbial cell surfaces. U(IV) was not associated with framboidal pyrite or nanometer-scale iron sulfides, which are presumed to be of microbial origin. Uranium concentrations were not detected in association with algal cells. Phylogenetic analyses of microbial populations in the sediment by the use of 16S rRNA and dissimilatory sulfite reductase gene sequences detected organisms belonging to the families Geobacteraceae and Desulfovibrionaceae. Cultivated members of these lineages reduce U(VI) and precipitate iron sulfides. The association of uranium with cells, but not with sulfide surfaces, suggests that U(VI) is reduced by the enzymatic activities of microorganisms. Uranium was highly enriched (760 ppm) in a subsurface black layer in unsaturated sediment sampled from a pit which was exposed to seasonal fluctuations in the pond level. XANES analysis showed that the majority of uranium in this layer was U(IV), indicating that uranium is preserved in its reduced form after burial. PMID- 15812003 TI - Agrobacterium tumefaciens-mediated transformation of Aspergillus fumigatus: an efficient tool for insertional mutagenesis and targeted gene disruption. AB - Agrobacterium tumefaciens was used to transform Aspergillus fumigatus by either random or site-directed integration of transforming DNA (T-DNA). Random mutagenesis via Agrobacterium tumefaciens-mediated transformation (ATMT) was accomplished with T-DNA containing a hygromycin resistance cassette. Cocultivation of A. fumigatus conidia and Agrobacterium (1:10 ratio) for 48 h at 24 degrees C resulted in high frequencies of transformation (> 100 transformants/10(7) conidia). The majority of transformants harbored a randomly integrated single copy of T-DNA and were mitotically stable. We chose alb1, a polyketide synthase gene, as the target gene for homologous integration because of the clear phenotype difference between the white colonies of Deltaalb1 mutant strains and the bluish-green colonies of wild-type strains. ATMT with a T-DNA containing alb1 disruption construct resulted in 66% albino transformants. Southern analysis revealed that 19 of the 20 randomly chosen albino transformants (95%) were disrupted by homologous recombination. These results suggest that ATMT is an efficient tool for transformation, random insertional mutagenesis, and gene disruption in A. fumigatus. PMID- 15812004 TI - Azospirillum brasilense produces the auxin-like phenylacetic acid by using the key enzyme for indole-3-acetic acid biosynthesis. AB - An antimicrobial compound was isolated from Azospirillum brasilense culture extracts by high-performance liquid chromatography and further identified by gas chromatography-mass spectrometry as the auxin-like molecule, phenylacetic acid (PAA). PAA synthesis was found to be mediated by the indole-3-pyruvate decarboxylase, previously identified as a key enzyme in indole-3-acetic acid (IAA) production in A. brasilense. In minimal growth medium, PAA biosynthesis by A. brasilense was only observed in the presence of phenylalanine (or precursors thereof). This observation suggests deamination of phenylalanine, decarboxylation of phenylpyruvate, and subsequent oxidation of phenylacetaldehyde as the most likely pathway for PAA synthesis. Expression analysis revealed that transcription of the ipdC gene is upregulated by PAA, as was previously described for IAA and synthetic auxins, indicating a positive feedback regulation. The synthesis of PAA by A. brasilense is discussed in relation to previously reported biocontrol properties of A. brasilense. PMID- 15812005 TI - Roles of four chitinases (chia, chib, chic, and chid) in the chitin degradation system of marine bacterium Alteromonas sp. strain O-7. AB - Alteromonas sp. strain O-7 secretes four chitinases (ChiA, ChiB, ChiC, and ChiD) in the presence of chitin. To elucidate why the strain produces multiple chitinases, we studied the expression levels of the four genes and proteins, their enzymatic properties, and their synergistic effects on chitin degradation. Among the four chitinases, ChiA was produced in the largest quantities, followed by ChiD, and the production of ChiB and ChiC changed at lower levels than those of ChiA and ChiD. The expression of the chiA, chiB, chiC, and chiD genes was investigated at the transcriptional level. The RNA transcript of chiA was most strongly induced in the presence of chitin, the expression of chiD followed, and the RNA transcripts of chiB and chiC changed at low levels. The hydrolyzing activities of the four chitinases against various substrates were examined. ChiA was the most active enzyme against powdered chitin, whereas ChiC was the most active against soluble chitin among the four chitinases. ChiD had activities closer to those of ChiA than to those of ChiB and ChiC. ChiB showed no distinctive feature against the chitinous substrates tested. When powdered chitin was treated with the proper combination of four chitinases, an approximately 2.0 fold increase in the hydrolytic activity was observed. These results, together with the results described above, indicate that ChiA plays a central role in chitin degradation for this strain. PMID- 15812006 TI - Anaerobic ammonia-oxidizing bacteria and related activity in Baltimore inner harbor sediment. AB - The discovery of bacteria capable of anaerobic ammonia oxidation (anammox) has generated interest in understanding the activity, diversity, and distribution of these bacteria in the environment. In this study anammox activity in sediment samples obtained from the Inner Harbor of Baltimore, Md., was detected by (15)N tracer assays. Anammox-specific oligonucleotide primer sets were used to screen a Planctomycetales-specific 16S rRNA gene library generated from sediment DNA preparations, and four new anammox bacterial sequences were identified. Three of these sequences form a cohesive new branch of the anammox group, and the fourth sequence branches separately from this group. Denaturing gradient gel electrophoresis analysis of sediment incubated with anammox-specific media confirmed the presence of the four anammox-related 16S rRNA gene sequences. Evidence for the presence of anammox bacteria in Inner Harbor sediment was also obtained by using an anammox-specific probe in fluorescence in situ hybridization studies. To our knowledge, this is the first report of anammox activity and related bacterial 16S rRNA gene sequences from the Chesapeake Bay basin area, and the results suggest that this pathway plays an important role in the nitrogen cycle of this estuarine environment. Furthermore, the presence of these bacteria and their activity in sediment strengthen the contention that anammox-related Plactomycetales are globally distributed. PMID- 15812007 TI - High-temperature-induced transposition of insertion elements in burkholderia multivorans ATCC 17616. AB - An efficient and quantitative method to analyze the transposition of various insertion sequence (IS) elements in Burkholderia multivorans ATCC 17616 was devised. pGEN500, a plasmid carrying a Bacillus subtilis-derived sacB gene, was introduced into ATCC 17616 cells, and 25% of their sucrose-resistant derivatives were found to carry various IS elements on pGEN500. A PCR-based experimental protocol, in which a mixture of several specific primer pairs was used, revealed that pGEN500 captured, in addition to five previously reported IS elements (IS401, IS402, IS406, IS407, and IS408), three novel IS elements, ISBmu1, ISBmu2, and ISBmu3. The global transposition frequency of these IS elements was enhanced more than sevenfold under a high-temperature condition (42 degrees C) but not under oxidative stress or starvation conditions. To our knowledge, this is the first report demonstrating the elevated transposition activities of several IS elements at a high temperature. The efficient experimental protocol developed in this study will be useful in quantitatively and simultaneously investigating various IS elements, as well as in capturing novel functional mobile elements from a wide variety of bacteria. PMID- 15812008 TI - Rapid engineering of the geldanamycin biosynthesis pathway by Red/ET recombination and gene complementation. AB - Genetic manipulation of antibiotic producers, such as Streptomyces species, is a rational approach to improve the properties of biologically active molecules. However, this can be a slow and sometimes problematic process. Red/ET recombination in an Escherichia coli host has permitted rapid and more versatile engineering of geldanamycin biosynthetic genes in a complementation plasmid, which can then be readily transferred into the Streptomyces host from which the corresponding wild type gene(s) has been removed. With this rapid Red/ET recombination and gene complementation approach, efficient gene disruptions and gene replacements in the geldanamycin biosynthetic gene cluster have been successfully achieved. As an example, we describe here the creation of a ketoreductase 6 null mutation in an E. coli high-copy-number plasmid carrying gdmA2A3 from Streptomyces hygroscopicus NRRL3602 and the subsequent complementation of a gdmA2A3 deletion host with this plasmid to generate a novel geldanamycin analog. PMID- 15812009 TI - Potential of the polyvalent anti-Staphylococcus bacteriophage K for control of antibiotic-resistant staphylococci from hospitals. AB - The increasing prevalence of antibiotic-resistant staphylococci has prompted the need for antibacterial controls other than antibiotics. In this study, a lytic bacteriophage (phage K) was assessed in vitro for its ability to inhibit emerging drug-resistant Staphylococcus aureus strains from hospitals and other species of Staphylococcus isolated from bovine infections. In in vitro inhibitory assays, phage K lysed a range of clinically isolated methicillin-resistant S. aureus (MRSA) strains, S. aureus with heterogeneous vancomycin resistance and vancomycin resistance, and teicoplanin-resistant strains. In these assays, 14 of the MRSA strains were initially only weakly sensitive to this phage. However, propagation of phage K on these less-sensitive strains resulted in all 14 being sensitive to the modified phages. The results enforce the principle that, while certain target bacteria may be relatively insensitive to lytic phage, this can be overcome by obtaining modified phage variants from passage of the phage through the insensitive strains. Model in situ hand wash studies using a phage-enriched wash solution resulted in a 100-fold reduction in staphylococcal numbers on human skin by comparison with numbers remaining after washing in phage-free solution. Infusion of the phage into a nonimmunogenic bismuth-based cream resulted in strong anti-Staphylococcus activity from the cream on plates and in broth. PMID- 15812010 TI - Exposure to cadmium elevates expression of genes in the OxyR and OhrR regulons and induces cross-resistance to peroxide killing treatment in Xanthomonas campestris. AB - Cadmium is an important heavy metal pollutant. For this study, we investigated the effects of cadmium exposure on the oxidative stress responses of Xanthomonas campestris, a soil and plant pathogenic bacterium. The exposure of X. campestris to low concentrations of cadmium induces cross-protection against subsequent killing treatments with either H2O2 or the organic hydroperoxide tert-butyl hydroperoxide (tBOOH), but not against the superoxide generator menadione. The cadmium-induced resistance to peroxides is due to the metal's ability to induce increased levels of peroxide stress protective enzymes such as alkyl hydroperoxide reductase (AhpC), monofunctional catalase (KatA), and organic hydroperoxide resistance protein (Ohr). Cadmium-induced resistance to H2O2 is dependent on functional OxyR, a peroxide-sensing transcription regulator. Cadmium induced resistance to tBOOH shows a more complex regulatory pattern. The inactivation of the two major sensor-regulators of organic hydroperoxide, OxyR and OhrR, only partially inhibited cadmium-induced protection against tBOOH, suggesting that these genes do have some role in the process. However, other, as yet unknown mechanisms are involved in inducible organic hydroperoxide protection. Furthermore, we show that the cadmium-induced peroxide stress response is mediated by the metal's ability to predominately cause an increase in intracellular concentrations of organic hydroperoxide and, in part, H2O2. Analyses of various mutants of peroxide-metabolizing enzymes suggested that this increase in organic hydroperoxide levels is, at least in part, responsible for cadmium toxicity in Xanthomonas. PMID- 15812011 TI - Biotinylation facilitates the uptake of large peptides by Escherichia coli and other gram-negative bacteria. AB - Gram-negative bacteria such as Escherichia coli can normally only take up small peptides less than 650 Da, or five to six amino acids, in size. We have found that biotinylated peptides up to 31 amino acids in length can be taken up by E. coli and that uptake is dependent on the biotin transporter. Uptake could be competitively inhibited by free biotin or avidin and blocked by the protonophore carbonyl m-chlorophenylhydrazone and was abolished in E. coli mutants that lacked the biotin transporter. Biotinylated peptides could be used to supplement the growth of a biotin auxotroph, and the transported peptides were shown to be localized to the cytoplasm in cell fractionation experiments. The uptake of biotinylated peptides was also demonstrated for two other gram-negative bacteria, Salmonella enterica serovar Typhimurium and Pseudomonas aeruginosa. This finding may make it possible to create new peptide antibiotics that can be used against gram-negative pathogens. Researchers have used various moieties to cause the illicit transport of compounds in bacteria, and this study demonstrates the illicit transport of the largest known compound to date. PMID- 15812012 TI - Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method. AB - The objective of this study was to develop and optimize a protocol for the rapid detection of Escherichia coli O157:H7 in aqueous samples by a combined immunomagnetic bead-immunoliposome (IMB/IL) fluorescence assay. The protocol consisted of the filtration or centrifugation of 30- to 100-ml samples followed by incubation of the filter membranes or pellet with anti-E. coli O157:H7 immunomagnetic beads in growth medium specific for E. coli O157:H7. The resulting E. coli O157:H7-immunomagnetic bead complexes were isolated by magnetic separation, washed, and incubated with sulforhodamine B-containing immunoliposomes specific for E. coli O157:H7; the final immunomagnetic bead-E. coli O157:H7-immunoliposome complexes were again isolated by magnetic separation, washed, and lysed with a n-octyl-beta-d-glucopyranoside to release sulforhodamine B. The final protocol took less than 8 h to complete and had a detection limit of less than 1 CFU of E. coli O157:H7 per ml in various aqueous matrices, including apple juice and cider. To validate the protocol at an independent facility, 100 ml samples of groundwater with and without E. coli O157:H7 (15 CFU) were analyzed by a public health laboratory using the optimized protocol and a standard microbiological method. While the IMB/IL fluorescence assay was able to identify E. coli O157:H7-containing samples with 100% accuracy, the standard microbiological method was unable to distinguish E. coli O157:H7-spiked samples from negative controls without further extensive workup. These results demonstrate the feasibility of using immunomagnetic beads in combination with sulforhodamine B-encapsulating immunoliposomes for the rapid detection of E. coli O157:H7 in aqueous samples. PMID- 15812013 TI - Nutrient effects on biocontrol of Penicillium roqueforti by Pichia anomala J121 during airtight storage of wheat. AB - The biocontrol yeast Pichia anomala inhibits the growth of a variety of mold species. We examined the mechanism underlying the inhibition of the grain spoilage mold Penicillium roqueforti by the biocontrol yeast P. anomala J121 during airtight storage. The biocontrol effect in a model grain silo with moist wheat (water activity of 0.96) was enhanced when complex medium, maltose, or glucose was added. Supplementation with additional nitrogen or vitamin sources did not affect the biocontrol activity of the yeast. The addition of complex medium or glucose did not significantly influence the yeast cell numbers in the silos, whether in the presence or absence of P. roqueforti. Mold growth was not influenced by the addition of nutrients, if cultivated without yeast. The products of glucose metabolism, mainly ethanol and ethyl acetate, increased after glucose addition to P. anomala-inoculated treatments. Our results suggest that neither competition for nutrients nor production of a glucose-repressible cell wall lytic enzyme is the main mode of action of biocontrol by P. anomala in this grain system. Instead, the mold-inhibiting effect probably is due to the antifungal action of metabolites, most likely a combination of ethyl acetate and ethanol, derived from glycolysis. The discovery that sugar amendments enhance the biocontrol effect of P. anomala suggests novel ways of formulating biocontrol yeasts. PMID- 15812014 TI - Rapid and sensitive detection of noroviruses by using TaqMan-based one-step reverse transcription-PCR assays and application to naturally contaminated shellfish samples. AB - Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples. PMID- 15812015 TI - Prevalence and genetic diversity of Campylobacter spp. in environmental water samples from a 100-square-kilometer predominantly dairy farming area. AB - Water samples were taken systematically from a 100-km2 area of mainly dairy farmland in northwestern England and examined for Campylobacter spp. Pulsed-field gel electrophoresis-restriction fragment length polymorphism (PFGE-RFLP) and flaA strain typing of Campylobacter jejuni and Campylobacter coli isolates were done. Data on the water source and the adjacent environment were recorded and examined as explanatory variables. Campylobacter spp. were isolated from 40.5% (n = 119) of the water samples tested. C. jejuni was isolated from 14.3%, C. coli was isolated from 18.5%, and Campylobacter lari was isolated from 4.2% of the samples. Campylobacter hyointestinalis was not isolated from any water source. The difference in prevalence between water types (trough, running, and standing) was significant (P = 0.001). C. jejuni was the species most commonly isolated from trough-water and running-water sources, while C. coli was the most frequently isolated from standing water (P < 0.001). No association was found between the presence of Escherichia coli and that of Campylobacter spp. The final multivariable logistic regression model for Campylobacter spp. included the following variables: water source, soil type, aspect, and amount of cattle fecal material in the environment (fecal pat count). Strain typing demonstrated a diverse population of C. jejuni and the presence of a common C. coli flaA type that was widely distributed throughout the area. Most of the isolates within the common flaA type were discriminated by PFGE-RFLP. These findings suggest a possible role for environmental water in the epidemiology of Campylobacter spp. in a farming environment. PMID- 15812016 TI - Folate cross-feeding supports symbiotic homoacetogenic spirochetes. AB - Treponema primitia, an H2-consuming CO2-reducing homoacetogenic spirochete in termite hindguts, requires an exogenous source of folate for growth. Tetrahydrofolate (THF) acts as a C1 carrier in CO2-reductive acetogenesis, a microbially mediated process important to the carbon and energy requirements of termites. To examine the hypothesis that other termite gut microbes probably supply some form of folate to T. primitia in situ, we used a bioassay to screen for and isolate folate-secreting bacteria from hindguts of Zootermopsis angusticollis, which is the host of T. primitia. Based on morphology, physiology, and 16S rRNA gene sequences, the major folate secretors were identified as strains of Lactococcus lactis and Serratia grimesii. During growth, these isolates secreted 5-formyl-THF at levels up to 146 ng/ml, and their cell-free culture fluids satisfied the folate requirement of T. primitia strains in vitro. Analysis of Z. angusticollis hindgut fluid revealed that 5-formyl-THF was the only detectable folate compound and occurred at an in situ concentration (1.3 mug/ml) which was more than sufficient to support the growth of T. primitia. These results imply that cross-feeding of 5-formyl-THF by other community members is important for growth of symbiotic hindgut spirochetes and thus termite nutrition and survival. PMID- 15812017 TI - Cytotoxicity potential and genotypic characterization of Escherichia coli isolates from environmental and food sources. AB - The presence of Escherichia coli isolates in the environment is a potential source of contamination of food and water supplies. Moreover, these isolates may harbor virulence genes that can be a source of new forms of pathogenic strains. Here, using multiplex PCR, we examined the presence of virulence gene markers (stx1, stx2, eaeA, hlyA) in 1,698 environmental isolates of E. coli and 81 isolates from food and clinical sources. The PCR analysis showed that approximately 5% (79 of 1,698) of the total environmental isolates and 96% (79 of 81) of the food and clinical isolates were positive for at least one of the genes. Of the food and clinical isolates, 84% (68 of 81 isolates) were positive for all four genes. Of the subset of environmental isolates chosen for further analysis, 16% (13 of 79 isolates) were positive for stx2 and 84% (66 of 79 isolates) were positive for eaeA; 16 of the latter strains were also positive for hlyA. The pathogenic potentials of 174 isolates (81 isolates from food and clinical sources and 93 isolates from environmental sources) were tested by using a cytotoxicity assay based on lactate dehydrogenase release from Vero cells. In general, 97% (79 of 81) of the food and clinical isolates and 41% (39 of 93) of the environmental isolates exhibited positive cytotoxicity. High cytotoxicity values correlated to the presence of stx genes. The majority of hly-positive but stx-negative environmental isolates also exhibited a certain degree of cytotoxicity. Isolates were also tested for sorbitol utilization and were genotyped by ribotyping and by repetitive extragenic palindromic PCR (REP-PCR) as potential means of quickly identifying virulent strains from the environment, but none of these methods could be used to distinguish cytotoxic environmental isolates. Only 31% of the isolates were negative for sorbitol fermentation, and none of the isolates had common ribotypes or REP-PCR fingerprints. This study suggests that overall higher cytotoxicity values correlated with the production of stx genes, and the majority of hly-positive but stx-negative environmental isolates also exhibited a certain degree of cytotoxicity. This study demonstrated that there is widespread distribution of potentially virulent E. coli strains in the environment that may be a cause of concern for human health. PMID- 15812018 TI - Genes involved in SkfA killing factor production protect a Bacillus subtilis lipase against proteolysis. AB - Small lipases of Bacillus species, such as LipA from Bacillus subtilis, have a high potential for industrial applications. Recent studies showed that deletion of six AT-rich islands from the B. subtilis genome results in reduced amounts of extracellular LipA. Here we demonstrate that the reduced LipA levels are due to the absence of four genes, skfABCD, located in the prophage 1 region. Intact skfABCD genes are required not only for LipA production at wild-type levels by B. subtilis 168 but also under conditions of LipA overproduction. Notably, SkfA has bactericidal activity and, probably, requires the SkfB to SkfD proteins for its production. The present results show that LipA is more prone to proteolytic degradation in the absence of SkfA and that high-level LipA production can be improved significantly by employing multiple protease-deficient B. subtilis strains. In conclusion, our findings imply that SkfA protects LipA, directly or indirectly, against proteolytic degradation. Conceivably, SkfA could act as a modulator in LipA folding or as a protease inhibitor. PMID- 15812019 TI - Cloning, expression, and site-directed mutagenesis of the propene monooxygenase genes from Mycobacterium sp. strain M156. AB - Propene monooxygenase has been cloned from Mycobacterium sp. strain M156, based on hybridization with the amoABCD genes of Rhodococcus corallinus B276. Sequencing indicated that the mycobacterial enzyme is a member of the binuclear nonheme iron monooxygenase family and, in gene order and sequence, is most similar to that from R. corallinus B-276. Attempts were made to express the pmoABCD operon in Escherichia coli and Mycobacterium smegmatis mc(2)155. In the former, there appeared to be a problem resolving overlapping reading frames between pmoA and -B and between pmoC and -D, while in the latter, problems were encountered with plasmid instability when the pmoABCD genes were placed under the control of the hsp60 heat shock promoter in the pNBV1 vector. Fortuitously, constructs with the opposite orientation were constitutively expressed at a level sufficient to allow preliminary mutational analysis. Two PMO active-site residues (A94 and V188) were targeted by site-directed mutagenesis to alter their stereoselectivity. The results suggest that changing the volume occupied by the side chain at V188 leads to a systematic alteration in the stereoselectivity of styrene oxidation, presumably by producing different orientations for substrate binding during catalysis. Changing the volume occupied by the side chain at A94 produced a nonsystematic change in stereoselectivity, which may be attributable to the role of this residue in expansion of the binding site during substrate binding. Neither set of mutations changed the enzyme's specificity for epoxidation. PMID- 15812020 TI - Formation of trans fatty acids is not involved in growth-linked membrane adaptation of Pseudomonas putida. AB - Fatty acid compositions in growing and resting cells of several strains of Pseudomonas putida (P8, NCTC 10936, and KT 2440) were studied, with a focus on alterations of the saturation degree, cis-trans isomerization, and cyclopropane formation. The fatty acid compositions of the strains were very similar under comparable growth conditions, but surprisingly, and contrary to earlier reports, trans fatty acids were not found in either exponentially growing cells or stationary-phase cells. During the transition from growth to the starvation state, cyclopropane fatty acids were preferentially formed, an increase in the saturation degree of fatty acids was observed, and larger amounts of hydroxy fatty acids were detected. A lowered saturation degree and concomitant higher membrane fluidity seemed to be optimal for substrate uptake and growth. The incubation of cells under nongrowth conditions rapidly led to the formation of trans fatty acids. We show that harvesting and sample preparation for analysis could provoke the enzyme-catalyzed formation of trans fatty acids. Freeze-thawing of resting cells and increased temperatures accelerated the formation of trans fatty acids. We demonstrate that cis-trans isomerization only occurred in cells that were subjected to an abrupt disturbance without having the possibility of adapting to the changed conditions by the de novo synthesis of fatty acids. The cis-trans isomerization reaction was in competition with the cis-to-cyclopropane fatty acid conversion. The potential for the formation of trans fatty acids depended on the cyclopropane content that was already present. PMID- 15812021 TI - Biphasic behavior of anammox regulated by nitrite and nitrate in an estuarine sediment. AB - The production of N2 gas via anammox was investigated in sediment slurries at in situ NO2- concentrations in the presence and absence of NO3-. With single enrichment above 10 microM 14NO2- or 14NO3- and 15NH4+, anammox activity was always linear (P < 0.05), in agreement with previous findings. In contrast, anammox exhibited a range of activity below 10 microM NO2- or NO3-, including an elevated response at lower concentrations. With 100 microM NO3-, no significant transient accumulation of NO2- could be measured, and the starting concentration of NO2- could therefore be regulated. With dual enrichment (1 to 20 microM NO2- plus 100 microM NO3-), there was a pronounced nonlinear response in anammox activity. Maximal activity occurred between 2 and 5 microM NO2-, but the amplitude of this peak varied across the study (November 2003 to June 2004). Anammox accounted for as much as 82% of the NO2- added at 1 microM in November 2003 but only for 15% in May 2004 and for 26 and 5% of the NO2- added at 5 microM for these two months, respectively. Decreasing the concentration of NO3- but holding NO2- at 5 microM decreased the significance of anammox as a sink for NO2 . The behavior of anammox was explored by use of a simple anammox-denitrification model, and the concept of a biphasic system for anammox in estuarine sediments is proposed. Overall, anammox is likely to be regulated by the availability of NO3- and NO2- and the relative size or activity of the anammox population. PMID- 15812022 TI - Diversity and phylogenetic affiliations of morphologically conspicuous large filamentous bacteria occurring in the pelagic zones of a broad spectrum of freshwater habitats. AB - Filamentous bacteria with a conspicuous morphology were found in the majority of the bacterioplankton samples from a variety of freshwater habitats that were studied. These heterotrophic filaments typically account for < 1 to 11% of the total number of bacteria. The biovolume of this morphotype can exceed 40% of the biovolume for all bacteria. Surprisingly, we found hardly any data on these morphologically conspicuous filaments in the literature. Mixed cultures containing these filamentous bacteria were established by cultivation and isolation experiments with samples from different freshwater lakes. Nearly full length 16S rRNA gene sequences were obtained from several mixed cultures and environmental samples from habitats in Europe, Africa, China, Australia, and New Zealand. Phylogenetic analysis of the sequences showed that three groups form a single monophyletic cluster, the SOL cluster, in the family Saprospiraceae. We developed a set of six nested probes for fluorescence in situ hybridization. Of the six probes, one probe was specific for Haliscomenobacter hydrossis, three probes were specific for the three subclusters (each probe was specific for one subcluster), one probe was specific for the entire SOL cluster, and another probe targeted almost the entire Saprospiraceae family. Specific hybridization of environmental samples and enrichments showed that the members of the three subclusters exhibited the same filamentous morphology. So far, using the subcluster-specific probes, we have not been able to detect any bacteria with a differing morphology. We conclude that the SOL cluster bacteria are an integral part of bacterioplankton in many freshwater habitats. They potentially account for a large fraction of the total bacterial biomass but have been underrepresented in molecular diversity studies so far. PMID- 15812023 TI - Elimination of botulinum neurotoxin (BoNT) type B from drinking water by small scale (personal-use) water purification devices and detection of BoNT in water samples. AB - Seven small-scale drinking water purification devices were evaluated for their capacity to eliminate botulinum neurotoxin (BoNT) type B from drinking water. Influent water inoculated with toxic Clostridium botulinum cultures and effluent purified water samples were tested for the presence of BoNT by using a standard mouse bioassay and two commercial rapid enzyme immunoassays (EIAs). The water purification devices based on filtration through ceramic or membrane filters with a pore size of 0.2 to 0.4 microm or irradiation from a low-pressure UV-lamp (254 nm) failed to remove BoNT from raw water (reduction of < 0.1 log10 units). A single device based on reverse osmosis was capable of removing the BoNT to a level below the detection limit of the mouse bioassay (reduction of > 2.3 log10 units). The rapid EIAs intended for the detection of BoNT from various types of samples failed to detect BoNT from aqueous samples containing an estimated concentration of BoNT of 396,000 ng/liter. PMID- 15812024 TI - Molecular detection of Mycobacterium bovis and Mycobacterium bovis BCG (Pasteur) in soil. AB - PCR primers specific for the Mycobacterium tuberculosis complex were used to detect the presence of Mycobacterium bovis BCG (Pasteur) in soil microcosms and Mycobacterium bovis in environmental samples taken from a farm in Ireland with a history of bovine tuberculosis. M. bovis genes were detected in soil at 4 and 21 months after possible contamination. Gene levels were found in the range of 1 x 10(3) to 3.6 x 10(3) gene copies g of soil(-1), depending on the sampling area. Areas around badger setts had the highest levels of detectable genes and were shown to have the highest levels of gene persistence. M. bovis-specific 16S rRNA sequences were detected, providing evidence of the presence of viable cells in Irish soils. Studies of DNA turnover in soil microcosms proved that dead cells of M. bovis BCG did not persist beyond 10 days. Further microcosm experiments revealed that M. bovis BCG survival was optimal at 37 degrees C with moist soil ( 20 kPa; 30% [vol/wt]). This study provides clear evidence that M. bovis can persist in the farm environment outside of its hosts and that climatic factors influence survival rates. PMID- 15812025 TI - Identification and molecular epidemiology of Campylobacter coli isolates from human gastroenteritis, food, and animal sources by amplified fragment length polymorphism analysis and Penner serotyping. AB - Campylobacter coli is an infrequently studied but important food-borne pathogen with a wide natural distribution. We investigated its molecular epidemiology by use of amplified fragment length polymorphism (AFLP)-based genotyping and Penner serotyping. Serotype reference strains and 177 Danish isolates of diverse origin identified by routine phenotyping as C. coli were examined. Molecular tools identified some 12% of field isolates as Campylobacter jejuni, emphasizing the need for improved identification methods in routine laboratories. Cluster analysis of AFLP profiles of 174 confirmed C. coli isolates revealed a difference in the distribution of isolates from pig and poultry (chicken, duck, turkey, and ostrich) species and indicated the various poultry species, but not pigs, to be likely sources of human C. coli infection. A poor correlation was observed between serotyping and AFLP profiling, suggesting that the former method has limited value in epidemiological studies of this species. PMID- 15812026 TI - Sec-mediated secretion of bacteriocin enterocin P by Lactococcus lactis. AB - Most lactic acid bacterium bacteriocins utilize specific leader peptides and dedicated machineries for secretion. In contrast, the enterococcal bacteriocin enterocin P (EntP) contains a typical signal peptide that directs its secretion when heterologously expressed in Lactococcus lactis. Signal peptide mutations and the SecA inhibitor azide blocked secretion. These observations demonstrate that EntP is secreted by the Sec translocase. PMID- 15812027 TI - Efficient production of L-Lactic acid by metabolically engineered Saccharomyces cerevisiae with a genome-integrated L-lactate dehydrogenase gene. AB - We developed a metabolically engineered yeast which produces lactic acid efficiently. In this recombinant strain, the coding region for pyruvate decarboxylase 1 (PDC1) on chromosome XII is substituted for that of the l-lactate dehydrogenase gene (LDH) through homologous recombination. The expression of mRNA for the genome-integrated LDH is regulated under the control of the native PDC1 promoter, while PDC1 is completely disrupted. Using this method, we constructed a diploid yeast transformant, with each haploid genome having a single insertion of bovine LDH. Yeast cells expressing LDH were observed to convert glucose to both lactate (55.6 g/liter) and ethanol (16.9 g/liter), with up to 62.2% of the glucose being transformed into lactic acid under neutralizing conditions. This transgenic strain, which expresses bovine LDH under the control of the PDC1 promoter, also showed high lactic acid production (50.2 g/liter) under nonneutralizing conditions. The differences in lactic acid production were compared among four different recombinants expressing a heterologous LDH gene (i.e., either the bovine LDH gene or the Bifidobacterium longum LDH gene): two transgenic strains with 2microm plasmid-based vectors and two genome-integrated strains. PMID- 15812028 TI - Production of novel rapamycin analogs by precursor-directed biosynthesis. AB - The natural product rapamycin, produced during fermentation by Streptomyces hygroscopicus, is known for its potent antifungal, immunosuppressive, and anticancer activities. During rapamycin biosynthesis, the amino acid l-pipecolate is incorporated into the rapamycin molecule. We investigated the use of precursor directed biosynthesis to create new rapamycin analogs by substitution of unusual l-pipecolate analogs in place of the normal amino acid. Our results suggest that the l-pipecolate analog (+/-)-nipecotic acid inhibits the biosynthesis of l pipecolate, thereby limiting the availability of this molecule for rapamycin biosynthesis. We used (+/-)-nipecotic acid in our precursor-directed biosynthesis studies to reduce l-pipecolate availability and thereby enhance the incorporation of other pipecolate analogs into the rapamycin molecule. We describe here the use of this method for production of two new sulfur-containing rapamycin analogs, 20 thiarapamycin and 15-deoxo-19-sulfoxylrapamycin, and report measurement of their binding to FKBP12. PMID- 15812029 TI - Culture-dependent and -independent methods to investigate the microbial ecology of Italian fermented sausages. AB - In this study, the microbial ecology of three naturally fermented sausages produced in northeast Italy was studied by culture-dependent and -independent methods. By plating analysis, the predominance of lactic acid bacteria populations was pointed out, as well as the importance of coagulase-negative cocci. Also in the case of one fermentation, the fecal enterocci reached significant counts, highlighting their contribution to the particular transformation process. Yeast counts were higher than the detection limit (> 100 CFU/g) in only one fermented sausage. Analysis of the denaturing gradient gel electrophoresis (DGGE) patterns and sequencing of the bands allowed profiling of the microbial populations present in the sausages during fermentation. The bacterial ecology was mainly characterized by the stable presence of Lactobacillus curvatus and Lactobacillus sakei, but Lactobacillus paracasei was also repeatedly detected. An important piece of evidence was the presence of Lactococcus garvieae, which clearly contributed in two fermentations. Several species of Staphylococcus were also detected. Regarding other bacterial groups, Bacillus sp., Ruminococcus sp., and Macrococcus caseolyticus were also identified at the beginning of the transformations. In addition, yeast species belonging to Debaryomyces hansenii, several Candida species, and Willopsis saturnus were observed in the DGGE gels. Finally, cluster analysis of the bacterial and yeast DGGE profiles highlighted the uniqueness of the fermentation processes studied. PMID- 15812030 TI - Expression of abrB310 and SinR, and effects of decreased abrB310 expression on the transition from acidogenesis to solventogenesis, in Clostridium acetobutylicum ATCC 824. AB - The transcription factors sinR and abrB are involved in the control of sporulation initiation in Bacillus subtilis. We identified a single homologue to sinR and three highly similar homologues to abrB, designated abrB310, abrB1941, and abrB3647, in Clostridium acetobutylicum ATCC 824. Using reporter vectors, we showed that the promoters of abrB1941 and abrB3647 were not active under the growth conditions tested. The abrB310 promoter was strongly active throughout growth and exhibited a transient elevation of expression at the onset of solventogenesis. Primer extension assays showed that two transcripts of abrB310 and a single, extremely weak transcript for sinR are expressed. Potential -35 and -10 consensus motifs are readily identifiable surrounding the transcription start sites of abrB310 and sinR, with a single putative 0A box present within the promoter of abrB310. In strains of C. acetobutylicum transformed with plasmids to elevate sinR expression or decrease sinR expression, no significant differences in growth or in acid or solvent production were observed compared to the control strains. In C. acetobutylicum strain 824(pAS310), which expressed an antisense RNA construct targeted against abrB310, the acids acetate and butyrate accumulated to approximately twice the normal concentration. This accumulation corresponded to a delay and decrease in acetone and butanol production. It was also found that sporulation in strain 824(pAS310) was delayed but that the morphology of sporulating cells and spores was normal. Based upon these observations, we propose that abrB310 may act as a regulator at the transition between acidogenic and solventogenic growth. PMID- 15812031 TI - Effects of artificial defoliation of pines on the structure and physiology of the soil fungal community of a mixed pine-spruce forest. AB - Loss of photosynthetic area can affect soil microbial communities by altering the availability of fixed carbon. We used denaturing gradient gel electrophoresis (DGGE) and Biolog filamentous-fungus plates to determine the effects of artificial defoliation of pines in a mixed pine-spruce forest on the composition of the fungal community in a forest soil. As measured by DGGE, two fungal species were affected significantly by the defoliation of pines (P < 0.001); the frequency of members of the ectomycorrhizal fungus genus Cenococcum decreased significantly, while the frequency of organisms of an unidentified soil fungus increased. The decrease in the amount of Cenococcum organisms may have occurred because of the formation of extensive hyphal networks by species of this genus, which require more of the carbon fixed by their host, or because this fungus is dependent upon quantitative differences in spruce root exudates. The defoliation of pines did not affect the overall composition of the soil fungal community or fungal-species richness (number of species per core). Biolog filamentous-fungus plate assays indicated a significant increase (P < 0.001) in the number of carbon substrates utilized by the soil fungi and the rate at which these substrates were used, which could indicate an increase in fungal-species richness. Thus, either small changes in the soil fungal community give rise to significant increases in physiological capabilities or PCR bias limits the reliability of the DGGE results. These data indicate that combined genetic and physiological assessments of the soil fungal community are needed to accurately assess the effect of disturbance on indigenous microbial systems. PMID- 15812032 TI - Diversity of transcripts of nitrite reductase genes (nirK and nirS) in rhizospheres of grain legumes. AB - Transcription of the nirK and nirS genes coding for dissimilatory bacterial nitrite reductases was analyzed by reverse transcription PCR (RT-PCR) of mRNA isolated from rhizosphere samples of three economically important grain legumes at maturity: Vicia faba, Lupinus albus, and Pisum sativum. The nirK gene and transcripts could be detected in all the rhizosphere samples. In contrast, nirS could not be detected. Sampling variations were analyzed by comparing denaturing gradient gel electrophoresis profiles derived from nirK RT-PCR products. High similarity was observed between the replicates, and so one representative product per legume was cloned. Clones with the correct insert size were screened by restriction fragment length polymorphism by using the restriction enzyme MspI. The clones could be distributed into 12 different patterns. Patterns 1, 3, 4, 5, and 7 were common in clone libraries of the three rhizosphere types under study. Patterns 2, 9, 10, and 11 were absent from Pisum rhizospheres, while patterns 6, 8, and 12 were absent from the Vicia library. Pattern 1, which was the most dominant in the Vicia and Lupinus libraries, constituted about 25% of all clones. The Lupinus library had clones representing all 12 patterns, indicating it to be the most diverse among the three. Clones representative of each pattern were sequenced. All patterns grouped together forming a distinct cluster, which was divergent from previously described nirK sequences in the database. The study revealed a hitherto unknown diversity of denitrifiers in legume rhizospheres. A plant-dependent rhizosphere effect on the transcripts of a gene was evident. PMID- 15812033 TI - Xanthomonas axonopodis pv. phaseoli var. fuscans is aggregated in stable biofilm population sizes in the phyllosphere of field-grown beans. AB - The occurrence of "Xanthomonas axonopodis pv. phaseoli var. fuscans" (proposed name) populations as biofilms on bean leaves was investigated during three field experiments on plots established with naturally contaminated bean seeds. Behavior of aggregated versus solitary populations was determined by quantification of culturable cells in different fractions of the epiphytic population separated by particle size. X. axonopodis pv. phaseoli var. fuscans population dynamic studies confirmed an asymptomatic and epiphytic colonization of the bean phyllosphere. For all years of experiment and cultivars tested, biofilms and solitary components of the populations were always detected. Biofilm population sizes remained stable throughout the growing season (around 10(5) CFU/g of fresh weight) while solitary population sizes were more abundant and varied with climate. According to enterobacterial repetitive intergenic consensus fingerprinting, aggregated bacterial isolates were not different from solitary isolates. In controlled conditions, application of a hydric stress resulted in a decrease of the solitary populations on the leaf surface while the biofilm fraction remained stable. Suppression of the hydric stress allowed solitary bacterial populations to increase again. Aggregation in biofilms on leaf surfaces provides protection to the bacterial cells against hydric stress. PMID- 15812035 TI - Improved assessment of denitrifying, N2-fixing, and total-community bacteria by terminal restriction fragment length polymorphism analysis using multiple restriction enzymes. AB - A database of terminal restriction fragments (tRFs) of the 16S rRNA gene was set up utilizing 13 restriction enzymes and 17,327 GenBank sequences. A computer program, termed TReFID, was developed to allow identification of any of these 17,327 sequences by means of polygons generated from the specific tRFs of each bacterium. The TReFID program complements and exceeds in its data content the Web based phylogenetic assignment tool recently described by A. D. Kent, D. J. Smith, B. J. Benson, and E. W. Triplett (Appl. Environ. Microb. 69:6768-6766, 2003). The method to identify bacteria is different, as is the region of the 16S rRNA gene employed in the present program. For the present communication the software of the tRF profiles has also been extended to allow screening for genes coding for N2 fixation (nifH) and denitrification (nosZ) in any bacterium or environmental sample. A number of controls were performed to test the reliability of the TReFID program. Furthermore, the TReFID program has been shown to permit the analysis of the bacterial population structure of bacteria by means of their 16S rRNA, nifH, and nosZ gene content in an environmental habitat, as exemplified for a sample from a forest soil. The use of the TReFID program reveals that noncultured denitrifying and dinitrogen-fixing bacteria might play a more dominant role in soils than believed hitherto. PMID- 15812034 TI - Diversity of oxygenase genes from methane- and ammonia-oxidizing bacteria in the Eastern Snake River Plain aquifer. AB - PCR amplification, restriction fragment length polymorphism, and phylogenetic analysis of oxygenase genes were used for the characterization of in situ methane and ammonia-oxidizing bacteria from free-living and attached communities in the Eastern Snake River Plain aquifer. The following three methane monooxygenase (MMO) PCR primer sets were used: A189-A682, which amplifies an internal region of both the pmoA gene of the MMO particulate form and the amoA gene of ammonia monooxygenase; A189-mb661, which specifically targets the pmoA gene; and mmoXA mmoXB, which amplifies the mmoX gene of the MMO soluble form (sMMO). Whole-genome amplification (WGA) was used to amplify metagenomic DNA from each community to assess its applicability for generating unbiased metagenomic template DNA. The majority of sequences in each archive were related to oxygenases of type II-like methanotrophs of the genus Methylocystis. A small subset of type I sequences found only in free-living communities possessed oxygenase genes that grouped nearest to Methylobacter and Methylomonas spp. Sequences similar to that of the amoA gene associated with ammonia-oxidizing bacteria (AOB) most closely matched a sequence from the uncultured bacterium BS870 but showed no substantial alignment to known cultured AOB. Based on these functional gene analyses, bacteria related to the type II methanotroph Methylocystis sp. were found to dominate both free living and attached communities. Metagenomic DNA amplified by WGA showed characteristics similar to those of unamplified samples. Overall, numerous sMMO like gene sequences that have been previously associated with high rates of trichloroethylene cometabolism were observed in both free-living and attached communities in this basaltic aquifer. PMID- 15812036 TI - Genetic diversity of benzoyl coenzyme A reductase genes detected in denitrifying isolates and estuarine sediment communities. AB - Benzoyl coenzyme A (benzoyl-CoA) reductase is a central enzyme in the anaerobic degradation of organic carbon, which utilizes a common intermediate (benzoyl-CoA) in the metabolism of many aromatic compounds. The diversity of benzoyl-CoA reductase genes in denitrifying bacterial isolates capable of degrading aromatic compounds and in river and estuarine sediment samples from the Arthur Kill in New Jersey and the Chesapeake Bay in Maryland was investigated. Degenerate primers were developed from the known benzoyl-CoA reductase genes from Thauera aromatica, Rhodopseudomonas palustris, and Azoarcus evansii. PCR amplification detected benzoyl-CoA reductase genes in the denitrifying isolates belonging to alpha-, beta-, or gamma-Proteobacteria as well as in the sediment samples. Phylogenetic analysis, sequence similarity comparison, and conserved indel determination grouped the new sequences into either the bcr type (found in T. aromatica and R. palustris) or the bzd type (found in A. evansii). All the Thauera strains and the isolates from the genera Acidovorax, Bradyrhizobium, Paracoccus, Ensifer, and Pseudomonas had bcr-type benzoyl-CoA reductases with amino acid sequence similarities of more than 97%. The genes detected from Azarocus strains were assigned to the bzd type. A total of 50 environmental clones were detected from denitrifying consortium and sediment samples, and 28 clones were assigned to either the bcr or the bzd type of benzoyl-CoA reductase genes. Thus, we could determine the genetic capabilities for anaerobic degradation of aromatic compounds in sediment communities of the Chesapeake Bay and the Arthur Kill on the basis of the detection of two types of benzoyl-CoA reductase genes. The detected genes have future applications as genetic markers to monitor aromatic compound degradation in natural and engineered ecosystems. PMID- 15812037 TI - Accumulation of polyhydroxyalkanoate from styrene and phenylacetic acid by Pseudomonas putida CA-3. AB - Pseudomonas putida CA-3 is capable of converting the aromatic hydrocarbon styrene, its metabolite phenylacetic acid, and glucose into polyhydroxyalkanoate (PHA) when a limiting concentration of nitrogen (as sodium ammonium phosphate) is supplied to the growth medium. PHA accumulation occurs to a low level when the nitrogen concentration drops below 26.8 mg/liter and increases rapidly once the nitrogen is no longer detectable in the growth medium. The depletion of nitrogen and the onset of PHA accumulation coincided with a decrease in the rate of substrate utilization and biochemical activity of whole cells grown on styrene, phenylacetic acid, and glucose. However, the efficiency of carbon conversion to PHA dramatically increased once the nitrogen concentration dropped below 26.8 mg/liter in the growth medium. When supplied with 67 mg of nitrogen/liter, the carbon-to-nitrogen (C:N) ratios that result in a maximum yield of PHA (grams of PHA per gram of carbon) for styrene, phenylacetic acid, and glucose are 28:1, 21:1, and 18:1, respectively. In cells grown on styrene and phenylacetic acid, decreasing the carbon-to-nitrogen ratio below 28:1 and 21:1, respectively, by increasing the nitrogen concentration and using a fixed carbon concentration leads to lower levels of PHA per cell and lower levels of PHA per batch of cells. Increasing the carbon-to-nitrogen ratio above 28:1 and 21:1 for cells grown on styrene and phenylacetic acid, respectively, by decreasing the nitrogen concentration and using a fixed carbon concentration increases the level of PHA per cell but results in a lower level of PHA per batch of cells. Increasing the carbon and nitrogen concentrations but maintaining the carbon-to-nitrogen ratio of 28:1 and 21:1 for cells grown on styrene and phenylacetic acid, respectively, results in an increase in the total PHA per batch of cells. The maximum yields for PHA from styrene, phenylacetic acid, and glucose are 0.11, 0.17, and 0.22 g of PHA per g of carbon, respectively. PMID- 15812038 TI - Vertical distribution of the free-living amoeba population in soil under desert shrubs in the Negev desert, Israel. AB - A field study was designed to examine the effect of desert shrubs on the dynamics of free-living amoebae in arid soil. Soil samples from 0- to 50-cm depths were collected at 10-cm intervals in each of the four seasons. The vertical distributions of the four main morphological types of amoebae, grouped according to their mobility, and of small flagellate populations were measured under the canopies of Hammada scoparia and Atriplex halimus, shrubs belonging to the chloride-absorbing xerohalophytes. The result obtained from the field study demonstrated that the total number of protozoa was significantly higher during the wet seasons (winter and spring) than during the dry seasons. The protozoan population was more diverse under the canopy of H. scoparia during the wet seasons, reaching 8,000 individuals per 1 g of dry soil, whereas during the dry seasons, the populations were higher under the canopy of A. halimus, with a mean of 250 individuals. The protozoan population in the deeper layers (40 to 50 cm) was found to be as active as that in the upper layers, demonstrating that, in the desert, soil columns below 20 cm are fertile and worth studying. The type 1 amoebae (e.g., Acanthamoeba and Filamoeba spp.) were the most abundant throughout the study period, and their numbers were significantly higher than those of the other amoeba types. PMID- 15812039 TI - Characterization of a mobile clpL gene from Lactobacillus rhamnosus. AB - Two genes encoding ClpL ATPase proteins were identified in a probiotic Lactobacillus rhamnosus strain, E-97800. Sequence analyses revealed that the genes, designated clpL1 and clpL2, share 80% identity. The clpL2 gene showed the highest degree of identity (98.5%) to a clpL gene from Lactobacillus plantarum WCFSI, while it was not detected in three other L. rhamnosus strains studied. According to Northern analyses, the expression of clpL1 and the clpL2 were induced during heat shock by > 20- and 3-fold, respectively. The functional promoter regions were determined by primer extension analyses, and the clpL1 promoter was found to be overlapped by an inverted repeat structure identical to the conserved CIRCE element, indicating that clpL1 belongs to the HrcA regulon in L. rhamnosus. No consensus binding sites for HrcA or CtsR could be identified in the clpL2 promoter region. Interestingly, the clpL2 gene was found to be surrounded by truncated transposase genes and flanked by inverted repeat structures nearly identical to the terminal repeats of the ISLpl1 from L. plantarum HN38. Furthermore, clpL2 was shown to be mobilized during prolonged cultivation at elevated temperature. The presence of a gene almost identical to clpL2 in L. plantarum and its absence in other L. rhamnosus strains suggest that the L. rhamnosus E-97800 has acquired the clpL2 gene via horizontal transfer. No change in the stress tolerance of the ClpL2-deficient derivative of E-97800 compared to the parental strain was observed. PMID- 15812040 TI - Molecular assays for targeting human and bovine enteric viruses in coastal waters and their application for library-independent source tracking. AB - Rapid population growth and urban development along waterways and coastal areas have led to decreasing water quality. To examine the effects of upstream anthropogenic activities on microbiological water quality, methods for source specific testing are required. In this study, molecular assays targeting human enteroviruses (HEV), bovine enteroviruses (BEV), and human adenoviruses (HAdV) were developed and used to identify major sources of fecal contamination in the lower Altamaha River, Georgia. Two-liter grab samples were collected monthly from five tidally influenced stations between July and December 2002. Samples were analyzed by reverse transcription- and nested-PCR. PCR results were confirmed by dot blot hybridization. Eleven and 17 of the 30 surface water samples tested positive for HAdV and HEV, respectively. Two-thirds of the samples tested positive for either HEV or HAdV, and the viruses occurred simultaneously in 26% of samples. BEV were detected in 11 of 30 surface water samples. Binary logistic regression analysis showed that the presence of both human and bovine enteric viruses was not significantly related to either fecal coliform or total coliform levels. The presence of these viruses was directly related to dissolved oxygen and streamflow but inversely related to water temperature, rainfall in the 30 days preceding sampling, and chlorophyll-a concentrations. The stringent host specificity of enteric viruses makes them good library-independent indicators for identification of water pollution sources. Viral pathogen detection by PCR is a highly sensitive and easy-to-use tool for rapid assessment of water quality and fecal contamination when public health risk characterization is not necessary. PMID- 15812041 TI - Spatial distribution and transcriptional activity of an uncultured clade of planktonic diazotrophic gamma-proteobacteria in the Arabian sea. AB - The spatial distribution of an uncultured clade of marine diazotrophic gamma proteobacteria in the Arabian Sea was investigated by the development of a specific primer pair to amplify an internal fragment of nifH by PCR. These organisms were most readily detected in highly oligotrophic surface waters but could also be found in deeper waters below the nutricline. nifH transcripts originating from this clade were detected in oligotrophic surface waters and, in addition, in the deeper and the more productive near-coastal waters. The nifH sequences most closely related to the unidentified marine bacterial group are from environmental clones amplified from the Atlantic and Pacific Oceans. These findings suggest that these gamma-proteobacteria are widespread and likely to be an important component of the heterotrophic diazotrophic microbial community of the tropical and subtropical oceans. PMID- 15812042 TI - Explorative multifactor approach for investigating global survival mechanisms of Campylobacter jejuni under environmental conditions. AB - Explorative approaches such as DNA microarray experiments are becoming increasingly important in microbial research. Despite these major technical advancements, approaches to study multifactor experiments are still lacking. We have addressed this problem by using rotation testing and a novel multivariate analysis of variance (MANOVA) approach (50-50 MANOVA) to investigate interacting experimental factors in a complex experimental design. Furthermore, a new rotation testing based method was introduced to calculate false-discovery rates for each response. This novel analytical concept was used to investigate global survival mechanisms in the environment of the major food-borne pathogen C. jejuni. We simulated nongrowth environmental conditions by investigating combinations of the factors temperature (5 and 25 degrees C) and oxygen tension (anaerobic, microaerobic, and aerobic). Data were generated with DNA microarrays for information about gene expression patterns and Fourier transform infrared (FT IR) spectroscopy to study global macromolecular changes in the cell. Microarray analyses showed that most genes were either unchanged or down regulated compared to the reference (day 0) for the conditions tested and that the 25 degrees C anaerobic condition gave the most distinct expression pattern with the fewest genes expressed. The few up-regulated genes were generally stress related and/or related to the cell envelope. We found, using FT-IR spectroscopy, that the amount of polysaccharides and oligosaccharides increased under the nongrowth survival conditions. Potential mechanisms for survival could be to down regulate most functions to save energy and to produce polysaccharides and oligosaccharides for protection against harsh environments. Basic knowledge about the survival mechanisms is of fundamental importance in preventing transmission of this bacterium through the food chain. PMID- 15812043 TI - Analysis of fluorescent protein expression in transformants of Rickettsia monacensis, an obligate intracellular tick symbiont. AB - We developed and applied transposon-based transformation vectors for molecular manipulation and analysis of spotted fever group rickettsiae, which are obligate intracellular bacteria that infect ticks and, in some cases, mammals. Using the Epicentre EZ::TN transposon system, we designed transposons for simultaneous expression of a reporter gene and a chloramphenicol acetyltransferase (CAT) resistance marker. Transposomes (transposon-transposase complexes) were electroporated into Rickettsia monacensis, a rickettsial symbiont isolated from the tick Ixodes ricinus. Each transposon contained an expression cassette consisting of the rickettsial ompA promoter and a green fluorescent protein (GFP) reporter gene (GFPuv) or the ompB promoter and a red fluorescent protein reporter gene (DsRed2), followed by the ompA transcription terminator and a second ompA promoter CAT gene cassette. Selection with chloramphenicol gave rise to rickettsial populations with chromosomally integrated single-copy transposons as determined by PCR, Southern blotting, and sequence analysis. Reverse transcription-PCR and Northern blots demonstrated transcription of all three genes. GFPuv transformant rickettsiae exhibited strong fluorescence in individual cells, but DsRed2 transformants did not. Western blots confirmed expression of GFPuv in R. monacensis and in Escherichia coli, but DsRed2 was expressed only in E. coli. The DsRed2 gene, but not the GFPuv gene, contains many GC-rich amino acid codons that are rare in the preferred codon suite of rickettsiae, possibly explaining the failure to express DsRed2 protein in R. monacensis. We demonstrated that our vectors provide a means to study rickettsia-host cell interactions by visualizing GFPuv-fluorescent R. monacensis associated with actin tails in tick host cells. PMID- 15812044 TI - Lipid biomarkers and carbon isotope signatures of a microbial (Beggiatoa) mat associated with gas hydrates in the gulf of Mexico. AB - White and orange mats are ubiquitous on surface sediments associated with gas hydrates and cold seeps in the Gulf of Mexico. The goal of this study was to determine the predominant pathways for carbon cycling within an orange mat in Green Canyon (GC) block GC 234 in the Gulf of Mexico. Our approach incorporated laser-scanning confocal microscopy, lipid biomarkers, stable carbon isotopes, and 16S rRNA gene sequencing. Confocal microscopy showed the predominance of filamentous microorganisms (4 to 5 mum in diameter) in the mat sample, which are characteristic of Beggiatoa. The phospholipid fatty acids extracted from the mat sample were dominated by 16:1omega7c/t (67%), 18:1omega7c (17%), and 16:0 (8%), which are consistent with lipid profiles of known sulfur-oxidizing bacteria, including Beggiatoa. These results are supported by the 16S rRNA gene analysis of the mat material, which yielded sequences that are all related to the vacuolated sulfur-oxidizing bacteria, including Beggiatoa, Thioploca, and Thiomargarita. The delta13C value of total biomass was -28.6 per thousand; those of individual fatty acids were -29.4 to -33.7 per thousand. These values suggested heterotrophic growth of Beggiatoa on organic substrates that may have delta13C values characteristic of crude oil or on their by-products from microbial degradation. This study demonstrated that integrating lipid biomarkers, stable isotopes, and molecular DNA could enhance our understanding of the metabolic functions of Beggiatoa mats in sulfide-rich marine sediments associated with gas hydrates in the Gulf of Mexico and other locations. PMID- 15812045 TI - Nonbioluminescent strains of Photobacterium phosphoreum produce the cell-to-cell communication signal N-(3-Hydroxyoctanoyl)homoserine lactone. AB - Bioluminescence is a common phenotype in marine bacteria, such as Vibrio and Photobacterium species, and can be quorum regulated by N-acylated homoserine lactones (AHLs). We extracted a molecule that induced a bacterial AHL monitor (Agrobacterium tumefaciens NT1 [pZLR4]) from packed cod fillets, which spoil due to growth of Photobacterium phosphoreum. Interestingly, AHLs were produced by 13 nonbioluminescent strains of P. phosphoreum isolated from the product. Of 177 strains of P. phosphoreum (including 18 isolates from this study), none of 74 bioluminescent strains elicited a reaction in the AHL monitor, whereas 48 of 103 nonbioluminescent strains did produce AHLs. AHLs were also detected in Aeromonas spp., but not in Shewanella strains. Thin-layer chromatographic profiles of cod extracts and P. phosphoreum culture supernatants identified a molecule similar in relative mobility (Rf value) and shape to N-(3-hydroxyoctanoyl)homoserine lactone, and the presence of this molecule in culture supernatants from a nonbioluminescent strain of P. phosphoreum was confirmed by high-performance liquid chromatography-positive electrospray high-resolution mass spectrometry. Bioluminescence (in a non-AHL-producing strain of P. phosphoreum) was strongly up regulated during growth, whereas AHL production in a nonbioluminescent strain of P. phosphoreum appeared constitutive. AHLs apparently did not influence bioluminescence, as the addition of neither synthetic AHLs nor supernatants delayed or reduced this phenotype in luminescent strains of P. phosphoreum. The phenotypes of nonbioluminescent P. phosphoreum strains regulated by AHLs remains to be elucidated. PMID- 15812046 TI - Rehydration of the lichen Ramalina lacera results in production of reactive oxygen species and nitric oxide and a decrease in antioxidants. AB - Lichens are slow-growing associations of fungi and unicellular green algae or cyanobacteria. They are poikilohydric organisms whose lifestyle in many cases consists of alternating periods of desiccation, with low metabolic activity, and hydration, which induces increase in their metabolism. Lichens have apparently adapted to such extreme transitions between desiccation and rehydration, but the mechanisms that govern these adaptations are still poorly understood. In this study, the effect of rehydration on the production of reactive oxygen species and nitric oxide as well as low-molecular-weight antioxidants was investigated with the lichen Ramalina lacera. Rehydration of R. lacera resulted in the initiation of and a rapid increase in photosynthetic activity. Recovery of photosynthesis was accompanied by bursts of intracellular production of reactive oxygen species and nitric oxide. Laser-scanning confocal microscopy using dichlorofluorescein fluorescence revealed that formation of reactive oxygen species following rehydration was associated with both symbiotic partners of the lichen. The rate and extent of reactive oxygen species production were similar in the light and in the dark, suggesting a minor contribution of photosynthesis. Diaminofluorescein fluorescence, indicating nitric oxide formation, was detected only in fungal hyphae. Activities associated with rehydration did not have a deleterious effect on membrane integrity as assessed by measurement of electrolyte leakage, but water-soluble low-molecular-weight antioxidants decreased significantly. PMID- 15812048 TI - New tool for metabolic pathway engineering in Escherichia coli: one-step method to modulate expression of chromosomal genes. AB - A simple and highly efficient method was developed to produce a library of Escherichia coli clones that express a particular chromosomal gene at a wide range of expression levels. The basic strategy was to replace all or part of the upstream region of a coding sequence containing the elements involved in its expression (promoter, operator, gene coding for a regulator, ribosome binding site, and start codon) with a PCR-generated library of expression cassettes. PMID- 15812047 TI - Mycobacterium avium subsp. paratuberculosis in the catchment area and water of the River Taff in South Wales, United Kingdom, and its potential relationship to clustering of Crohn's disease cases in the city of Cardiff. AB - In South Wales, United Kingdom, a populated coastal region lies beneath hill pastures grazed by livestock in which Mycobacterium avium subsp. paratuberculosis is endemic. The Taff is a spate river running off the hills and through the principal city of Cardiff. We sampled Taff water above Cardiff twice weekly from November 2001 to November 2002. M. avium subsp. paratuberculosis was detected by IS900 PCR and culture. Thirty-one of 96 daily samples (32.3%) were IS900 PCR positive, and 12 grew M. avium subsp. paratuberculosis bovine strains. Amplicon sequences from colonies were identical to the sequence with GenBank accession no. X16293, whereas 16 of 19 sequences from river water DNA extracts had a single nucleotide polymorphism at position 214. This is consistent with a different strain of M. avium subsp. paratuberculosis in the river, which is unculturable by the methods we used. Parallel studies showed that M. avium subsp. paratuberculosis remained culturable in lake water microcosms for 632 days and persisted to 841 days. Of four reservoirs controlling the catchment area of the Taff, M. avium subsp. paratuberculosis was present in surface sediments from three and in sediment cores from two, consistent with deposition over at least 50 years. Previous epidemiological research in Cardiff demonstrated a highly significant increase of Crohn's disease in 11 districts. These bordered the river except for a gap on the windward side. A topographical relief map shows that this gap is directly opposite a valley open to the prevailing southwesterly winds. This would influence the distribution of aerosols carrying M. avium subsp. paratuberculosis from the river. PMID- 15812049 TI - Novel major bacterial candidate division within a municipal anaerobic sludge digester. AB - In a previous study, we analyzed the molecular diversity of Planctomycetales by PCR amplification and sequencing of 16S rRNA clone libraries generated from a municipal wastewater plant, using planctomycete-specific and universal primer sets (R. Chouari, D. Le Paslier, P. Daegelen, P. Ginestet, J. Weissenbach, and A. Sghir, Appl. Environ. Microbiol. 69:7354-7363, 2003). Only a small fraction (4%) of the 16S rRNA gene sequences of the digester clone library corresponded to the Planctomycetales division. Importantly, 85.9% of the digester clone sequences are grouped into two different clusters named WWE1 (81.4% of the sequences) and WWE2 (4.5%) and are distantly affiliated with unidentified bacterial sequences retrieved from a methanogenic reactor community and from a termite gut, respectively. In phylogenetic analysis using 16S rRNA gene sequence representatives of the main phylogenetic bacterial divisions, the two clusters are monophyletic, branch apart from each other, and are distantly related to Planctomycetales and other bacterial divisions. A novel candidate division is proposed for WWE1, while the WWE2 cluster strongly affiliates with the recently proposed Lentisphearae phylum. We designed and validated a 16S rRNA probe targeting WWE1 16S rRNA sequences by both fluorescent in situ hybridization (FISH) and dot blot hybridization (DBH). Results of FISH analysis show that WWE1 representative microorganisms are rods or filamentous shaped, while DBH shows that WWE1 accounts for 12% of the total bacterial rRNA within the anaerobic digester. The remaining 16S rRNA gene sequences are affiliated with Verrucomicrobia or recently described candidate divisions with no known pure culture representatives, such as OD1, BRC1, or NBL-UPA2, making up less than 3.5% of the clone library, respectively. This inventory expands the known diversity of the latter bacterial division-level lineages. PMID- 15812050 TI - Differential sunlight sensitivity of picophytoplankton from surface Mediterranean Coastal Waters. AB - We tested the sensitivity of coastal picophytoplankton exposed to natural sunlight in short-term experiments. Cell abundance and cell-specific chlorophyll fluorescence were significantly reduced in Prochlorococcus spp. but not in Synechococcus, whereas picoeukaryotes had an intermediate response. These results are the first direct evidence of a differential sensitivity to sunlight of these ubiquitous marine members of unicellular phytoplankton. PMID- 15812051 TI - Detection of Salmonella strains and Escherichia coli O157:H7 in feces of small ruminants and their isolation with various media. AB - Salmonella strains and Escherichia coli O157:H7 were detected in 17 and 5 small ruminants in Virginia, respectively, of 287 tested. Background microflora interfered with the fecal analysis. The combination of Salmonella enzyme immunoassay (EIA) detection and xylose-lysine-deoxycholate agar isolation was satisfactory. Modifying enrichment to a 1:100 dilution enabled effective E. coli O157:H7 detection by EIA and isolation by sorbitol-MacConkey agar with cefixime tellurite. PMID- 15812053 TI - Anaerobic, nitrate-dependent oxidation of U(IV) oxide minerals by the chemolithoautotrophic bacterium Thiobacillus denitrificans. AB - Under anaerobic conditions and at circumneutral pH, cells of the widely distributed, obligate chemolithoautotrophic bacterium Thiobacillus denitrificans oxidatively dissolved synthetic and biogenic U(IV) oxides (uraninite) in nitrate dependent fashion: U(IV) oxidation required the presence of nitrate and was strongly correlated with nitrate consumption. This is the first report of anaerobic U(IV) oxidation by an autotrophic bacterium. PMID- 15812052 TI - Comparative analysis of bacterial diversity in freshwater sediment of a shallow eutrophic lake by molecular and improved cultivation-based techniques. AB - Comparative analysis of bacterial diversity in freshwater sediment collected from a shallow eutrophic lake was performed by using 16S rRNA gene clone library and improved cultivation-based techniques. Our study demonstrated that the use of gellan gum as a gelling reagent instead of agar was more effective at increasing culturability, cultivating a diverse array of novel microbes, and reducing the gaps of the results between molecular and cultivation-based analyses. PMID- 15812054 TI - Viscoelasticity of Staphylococcus aureus biofilms in response to fluid shear allows resistance to detachment and facilitates rolling migration. AB - Staphylococcus aureus is a leading cause of catheter-related bloodstream infections and endocarditis. Both involve (i) biofilm formation, (ii) exposure to fluid shear, and (iii) high rates of dissemination. We found that viscoelasticity allowed S. aureus biofilms to resist detachment due to increased fluid shear by deformation, while remaining attached to a surface. Further, we report that S. aureus microcolonies moved downstream by rolling along the lumen walls of a glass flow cell, driven by the flow of the overlying fluid. The rolling appeared to be controlled by viscoelastic tethers. This tethered rolling may be important for the surface colonization of medical devices by nonmotile bacteria. PMID- 15812055 TI - Characterization of the export of bulk poly(A)+ mRNA in Saccharomyces cerevisiae during the wine-making process. AB - Ethanol stress affects the nuclear export of mRNA similarly to heat shock in Saccharomyces cerevisiae. However, we have little information about mRNA transport in actual alcoholic fermentation. Here we characterized the transport of mRNA during wine making and found that bulk poly(A)+ mRNA accumulated in the nucleus as fermentation progressed. PMID- 15812057 TI - Evidence for involvement of an electron shuttle in electricity generation by Geothrix fermentans. AB - In experiments performed using graphite electrodes poised by a potentiostat (+200 mV versus Ag/AgCl) or in a microbial fuel cell (with oxygen as the electron acceptor), the Fe(III)-reducing organism Geothrix fermentans conserved energy to support growth by coupling the complete oxidation of acetate to reduction of a graphite electrode. Other organic compounds, such as lactate, malate, propionate, and succinate as well as components of peptone and yeast extract, were utilized for electricity production. However, electrical characteristics and the results of shuttling assays indicated that unlike previously described electrode-reducing microorganisms, G. fermentans produced a compound that promoted electrode reduction. This is the first report of complete oxidation of organic compounds linked to electrode reduction by an isolate outside of the Proteobacteria. PMID- 15812056 TI - Degradation of beta-Hexachlorocyclohexane by Haloalkane Dehalogenase LinB from Sphingomonas paucimobilis UT26. AB - Beta-Hexachlorocyclohexane (beta-HCH) is the most recalcitrant among the alpha-, beta-, gamma-, and delta-isomers of HCH and causes serious environmental pollution problems. We demonstrate here that the haloalkane dehalogenase LinB, reported earlier to mediate the second step in the degradation of gamma-HCH in Sphingomonas paucimobilis UT26, metabolizes beta-HCH to produce 2,3,4,5,6 pentachlorocyclohexanol. PMID- 15812058 TI - Quantitative detection of Listeria monocytogenes in biofilms by real-time PCR. AB - A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 x 10(2) CFU/cm2. PMID- 15812059 TI - Pathways for methanogenesis and diversity of methanogenic archaea in three boreal peatland ecosystems. AB - The main objectives of this study were to uncover the pathways used for methanogenesis in three different boreal peatland ecosystems and to describe the methanogenic populations involved. The mesotrophic fen had the lowest proportion of CH4 produced from H2-CO2. The oligotrophic fen was the most hydrogenotrophic, followed by the ombrotrophic bog. Each site was characterized by a specific group of methanogenic sequences belonging to Methanosaeta spp. (mesotrophic fen), rice cluster-I (oligotrophic fen), and fen cluster (ombrotrophic bog). PMID- 15812060 TI - Opposite enantioselectivities of two phenotypically and genotypically similar strains of Pseudomonas frederiksbergensis in bacterial whole-cell sulfoxidation. AB - Soil samples were screened to select microorganisms with the capability to oxidize organic sulfides into the corresponding sulfoxides with differential enantioselectivities. Several bacterial strains that preferentially produced the S-configured sulfoxide enantiomer were isolated. Surprisingly, one bacterial strain, genotypically and phenotypically characterized as Pseudomonas frederiksbergensis, selectively gave the R enantiomer. The finding that two apparently identical organisms displayed opposite enantioselectivities is novel for non-genetically modified organisms. PMID- 15812061 TI - Presence of norovirus sequences in bottled waters is questionable. PMID- 15812062 TI - Possible errors in the analysis of lactic acid and volatile fatty acids in the gastrointestinal tracts of pigs and chickens. PMID- 15812063 TI - Second biannual report of the Cochrane Haematological Malignancies Group. PMID- 15812064 TI - An apple a day...does it really keep the doctor away? The current state of cancer chemoprevention. PMID- 15812065 TI - Progestins and recurrence in breast cancer survivors. PMID- 15812066 TI - FDA Oncology Committee debates Iressa's status following negative trial results. PMID- 15812067 TI - Recent studies show limited association of fruit and vegetable consumption and cancer risk. PMID- 15812068 TI - An integrated approach: systems biology seeks order in complexity. PMID- 15812069 TI - Stat bite: Incidence of brain and other nervous system cancers, 1975-2001. PMID- 15812070 TI - Large variations seen in treatment of adults with brain cancer. PMID- 15812071 TI - Trial quickly changed management of cervical abnormalities. PMID- 15812073 TI - A randomized trial of antioxidant vitamins to prevent second primary cancers in head and neck cancer patients. AB - BACKGROUND: Although low dietary intakes of antioxidant vitamins and minerals have been associated with higher risks of cancer, results of trials testing antioxidant supplementation for cancer chemoprevention have been equivocal. We assessed whether supplementation with antioxidant vitamins could reduce the incidence of second primary cancers among patients with head and neck cancer. METHODS: We conducted a multicenter, double-blind, placebo-controlled, randomized chemoprevention trial among 540 patients with stage I or II head and neck cancer treated by radiation therapy between October 1, 1994, and June 6, 2000. Supplementation with alpha-tocopherol (400 IU/day) and beta-carotene (30 mg/day) or placebo began on the first day of radiation therapy and continued for 3 years after the end of radiation therapy. In the course of the trial, beta-carotene supplementation was discontinued after 156 patients had enrolled because of ethical concerns. The remaining patients received alpha-tocopherol or placebo only. Survival was evaluated by Kaplan-Meier analysis. Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). All statistical tests were two-sided. RESULTS: After a median follow-up of 52 months, second primary cancers and recurrences of the first tumor were diagnosed in 113 and 119 participants, respectively. The effect of supplementation on the incidence of second primary cancers varied over time. Compared with patients receiving placebo, patients receiving alpha-tocopherol supplements had a higher rate of second primary cancers during the supplementation period (HR = 2.88, 95% CI = 1.56 to 5.31) but a lower rate after supplementation was discontinued (HR = 0.41, 95% CI = 0.16 to 1.03). Similarly, the rate of having a recurrence or second primary cancer was higher during (HR = 1.86, 95% CI = 1.27 to 2.72) but lower after (HR = 0.71, 95% CI = 0.33 to 1.53) supplementation with alpha-tocopherol. The proportion of participants free of second primary cancer overall after 8 years of follow-up was similar in both arms. CONCLUSIONS: alpha-Tocopherol supplementation produced unexpected adverse effects on the occurrence of second primary cancers and on cancer-free survival. PMID- 15812074 TI - Recombinant human erythropoietin and overall survival in cancer patients: results of a comprehensive meta-analysis. AB - BACKGROUND: Anemia associated with cancer and cancer therapy is an important clinical and economic factor in the treatment of malignant diseases. METHODS: We conducted a systematic literature review to assess the efficacy of erythropoietin to prevent or treat anemia in cancer patients with regard to red blood cell transfusions, hematologic response, adverse events, and overall survival. We searched the Cochrane Library, Medline, EMBASE, and other databases for relevant articles published from January 1985 to December 2001. We included all randomized controlled trials that compared the use of recombinant human erythropoietin (plus transfusion, if needed) with no erythropoietin treatment (plus transfusion, if needed). Relative risks (RRs) and 95% confidence intervals (CIs) were calculated under a fixed-effects model. Clinical and statistical heterogeneity were examined with sensitivity analyses and meta-regression. Statistical tests for effect estimates were two-sided. RESULTS: We identified 27 trials involving 3287 adult patients. Patients treated with erythropoietin had a lower relative risk of having a blood transfusion than untreated patients (RR = 0.67, 95% CI = 0.62 to 0.73). Erythropoietin-treated patients with baseline hemoglobin levels lower than 10 g/dL were more likely to have a hematologic response than untreated patients (RR = 3.60, 95% CI = 3.07 to 4.23). The relative risk for thromboembolic complications after erythropoietin treatment was not statistically significantly increased (RR = 1.58, 95% CI = 0.94 to 2.66) compared with that of untreated patients. There is suggestive but inconclusive evidence that erythropoietin may improve overall survival (adjusted data: hazard ratio [HR] = 0.81, 95% CI = 0.67 to 0.99; unadjusted data: HR = 0.84, 95% CI = 0.69 to 1.02). CONCLUSIONS: Erythropoietin treatment may reduce the risk for blood transfusions and improve hematologic response in cancer patients. However, our favorable survival outcome is in contrast to two large (N = 351 and 939) recently published randomized controlled trials in which erythropoietin-treated patients had statistically significantly worse survival than untreated patients. Possible reasons for the disparity with our results include differences in study population and design, higher target hemoglobin levels and higher risk of thromboembolic complications, and concerns that erythropoietin may stimulate tumor growth. PMID- 15812075 TI - Randomized study of maintenance vinorelbine in responders with advanced non-small cell lung cancer. AB - BACKGROUND: Prolongation of chemotherapy duration, usually referred to as maintenance chemotherapy, has been considered as an approach to improve survival of patients with advanced non-small-cell lung cancer (NSCLC). If the maintenance regimen differs from the induction regimen, patients will receive not only higher total doses of chemotherapy but also earlier delivery of non-cross-resistant agents. We conducted a randomized trial to compare maintenance vinorelbine therapy with observation in previously untreated patients who responded to induction treatment with mitomycin-ifosfamide-cisplatin (MIC). METHODS: Patients with stage IIIB NSCLC were treated with two monthly MIC cycles followed by radiotherapy; those with "wet" stage IIIB (pleural or pericardial involvement), with stage IIIB with supraclavicular node involvement, or stage IV (i.e., metastatic) NSCLC were treated with four monthly MIC cycles. Patients who responded to induction treatment were randomly assigned to receive intravenous vinorelbine at a dose of 25 mg x m(-2) x wk(-1) for 6 months or no further treatment. Survival comparisons used the log-rank test and the Cox regression adjusted for stage. All statistical tests were two-sided. RESULTS: A total of 573 patients were registered, of whom 227 responded to induction treatment and 181 were randomly assigned (91 to maintenance vinorelbine and 90 to observation) between January 1994 and March 2000. One- and 2-year survival rates were 42.2% and 20.1% in the vinorelbine arm and 50.6% and 20.2% in the observation arm, respectively (log-rank P = .48). The hazard ratio of survival after adjustment on stage, in the vinorelbine arm relative to the observation arm, was 1.08 (95% confidence interval = 0.79 to 1.47; P = .65). There was also no difference between arms in progression-free survival (log-rank P = .32). CONCLUSION: Maintenance vinorelbine did not improve survival of patients with advanced NSCLC who responded to induction MIC treatment. Nevertheless, other agents, including docetaxel and targeted agents, should be evaluated as maintenance agents before the concept is abandoned. PMID- 15812076 TI - Effects of light and food schedules on liver and tumor molecular clocks in mice. AB - BACKGROUND: Disrupted circadian coordination accelerates malignant growth, but the molecular mechanism is unclear. METHODS: Healthy or Glasgow osteosarcoma bearing mice (n = 162) were synchronized with light and darkness over 2-3 weeks, submitted to an 8-hour advance onset of light every 2 days (chronic jet lag) to disrupt circadian coordination, or submitted to chronic jet lag and meal timing to prevent molecular clock alteration. The expression of molecular clock genes and of the cell cycle genes c-Myc and p53 in liver and tumor was determined with quantitative reverse transcription-polymerase chain reaction at six circadian times over a 24-hour period of light and darkness and analyzed with analysis of variance and cosinor. Tumor weight was measured daily over the course of the experiment. All statistical tests were two-sided. RESULTS: In synchronized mice, mean mRNA levels of clock genes Rev-erbalpha, Per2, and Bmal1 varied by 206-, four-, and 26-fold, respectively, over the 24 hours in healthy mouse liver; by 36 , 35-, and 32-fold in the livers of tumor-bearing mice; and by 9.4-, 5.5-, and sixfold in tumor tissue (P = .046 to <.001). In mice subjected to chronic jet lag, the periodic changes were dampened and the clock gene rhythms were temporally shifted in liver and ablated in tumor, and tumor growth was accelerated. Meal timing reversed the chronic jet lag-induced alterations in Rev erbalpha and Per2 expression in liver and of all three clock genes in tumor and slowed tumor growth. Tumor growth differed as a function of light and feeding schedules (P = .04). No obvious rhythm was detected for p53 or c-Myc in liver or tumor tissues of synchronized mice. In healthy mice subjected to chronic jet lag, the mean level of p53 expression was cut in half (P = .002), and a 12-fold circadian variation in c-Myc mRNA level (P = .03) was induced in the liver of healthy mice, whereas complex expression patterns were found in the liver and tumor of tumor-bearing mice. CONCLUSIONS: Altered light-dark or feeding schedules modified the expression of molecular clock genes and genes involved in carcinogenesis and tumor progression. PMID- 15812077 TI - Dietary patterns and pancreatic cancer risk in men and women. AB - BACKGROUND: Diabetes appears to be associated with the development of pancreatic cancer. Three large prospective cohort studies observed a statistically significant relation between obesity and pancreatic cancer risk. Dietary patterns have been associated with fasting insulin levels and risk of diabetes. To determine whether dietary patterns are associated with pancreatic cancer risk, we analyzed data from two large prospective cohort studies. METHODS: We combined data for men and women to obtain a total of 366 cases of incident pancreatic cancer from a total of 124,672 eligible participants. Dietary data were obtained from food frequency questionnaires in 1986 for men and in 1984 for women. We identified two major dietary patterns, prudent and western, by factor analysis. The prudent pattern was characterized by high fruit and vegetable intake; the western pattern was characterized by high meat and high fat intakes. Multivariable relative risks (RRs) were adjusted for potential confounders, including smoking and body mass index. RESULTS: In the pooled analysis of men and women, no associations were observed between the prudent pattern (RR = 1.32, 95% confidence interval [CI] = 0.66 to 2.63, for highest versus lowest quintile) or the western pattern (RR = 0.91, 95% CI = 0.57 to 1.47, for highest versus lowest quintile) and the risk of pancreatic cancer. Stratifying by body mass index or physical activity did not change the associations. CONCLUSION: Dietary patterns were not associated with the risk of pancreatic cancer in two large cohort studies of men and women. PMID- 15812078 TI - Sequence variants in Toll-like receptor gene cluster (TLR6-TLR1-TLR10) and prostate cancer risk. AB - BACKGROUND: Chronic inflammation plays an important role in several human cancers and may be involved in the etiology of prostate cancer. Toll-like receptors (TLRs) are important in the innate immune response to pathogens and in cross-talk between innate immunity and adaptive immunity. Our previous finding of an association of TLR4 gene sequence variants and prostate cancer risk provides evidence for a role of TLRs in prostate cancer. In this study, we investigated whether sequence variants in the TLR6-TLR1-TLR10 gene cluster, residing within a 54-kb region on 4p14, were associated with prostate cancer risk. METHODS: We selected 32 single-nucleotide polymorphisms (SNPs) covering these three genes and genotyped these SNPs in 96 control subjects from the Cancer Prostate in Sweden (CAPS) population-based prostate cancer case-control study. Five distinct haplotype blocks were inferred at this region, and we identified 17 haplotype tagging SNPs (htSNPs) that could uniquely describe >95% of the haplotypes. These 17 htSNPs were then genotyped in the entire CAPS study population (1383 case subjects and 780 control subjects). Odds ratios of prostate cancer for the carriers of a variant allele versus those with the wild-type allele were estimated using unconditional logistic regression. RESULTS: The allele frequencies of 11 of the 17 SNPs were statistically significantly different between case and control subjects (P = .04-.001), with odds ratios for variant allele carriers (homozygous or heterozygous) compared with wild-type allele carriers ranging from 1.20 (95% confidence interval [CI] = 1.00 to 1.43) to 1.38 (95% CI = 1.12 to 1.70). Phylogenetic tree analyses of common haplotypes identified a clade of two evolutionarily related haplotypes that are statistically significantly associated with prostate cancer risk. These two haplotypes contain all the risk alleles of these 11 associated SNPs. CONCLUSION: The observed multiple associated SNPs at the TLR6-TLR1-TLR10 gene cluster were dependent and suggest the presence of a founder prostate cancer risk variant on this haplotype background. The TLR6-TLR1-TLR10 gene cluster may play a role in prostate cancer risk, although further functional studies are needed to pinpoint the disease-associated variants in this gene cluster. PMID- 15812079 TI - Menopausal hormone therapy after breast cancer: the Stockholm randomized trial. AB - In 1997 two independent randomized clinical trials, Hormonal Replacement Therapy After Breast Cancer--Is It Safe? (HABITS; 434 patients) and the Stockholm trial (378 patients), were initiated in Sweden to compare menopausal hormone therapy with no menopausal hormone therapy after diagnosis of early-stage breast cancer. Much of the design of both studies was similar; however, a goal of the Stockholm protocol, not shared with the HABITS trial, was to minimize the use of progestogen combined with estrogen. The HABITS trial was prematurely stopped in December 2003, because, at a median follow-up of 2.1 years, the risk for recurrence of breast cancer among patients receiving menopausal hormone therapy was statistically significantly higher (relative hazard [RH] = 3.3, 95% confidence interval [CI] = 1.5 to 7.4) than among those receiving no treatment. In the Stockholm trial, however, at a median follow-up of 4.1 years, the risk of breast cancer recurrence was not associated with menopausal hormone therapy (RH = 0.82, 95% CI = 0.35 to 1.9). Statistically significant heterogeneity in the rate of recurrence was observed (P = .02; two-sided likelihood-ratio test) between the two studies, indicating that chance may not be the only explanation. Doses of estrogen and progestogen and treatment regimens for menopausal hormone therapy may be associated with the recurrence of breast cancer. PMID- 15812080 TI - Overall survival after concurrent cisplatin-radiotherapy compared with radiotherapy alone in locoregionally advanced nasopharyngeal carcinoma. AB - This phase III randomized study compared concurrent cisplatin-radiotherapy (CRT) versus radiotherapy (RT) alone in patients with locoregionally advanced nasopharyngeal carcinoma. A total of 350 patients were randomly assigned to receive external RT alone or concurrently with cisplatin at a dosage of 40 mg/m(2) weekly. The primary endpoint was overall survival, and the median follow up was 5.5 years. The 5-year overall survival was 58.6% (95% confidence interval [CI] = 50.9% to 66.2%) for the RT arm and 70.3% (95% CI = 63.4% to 77.3%) for the CRT arm. In Cox regression analysis adjusted for T stage, age, and overall stage, the difference in overall survival was statistically significantly in favor of concurrent CRT (P = .049, hazard ratio [HR] = 0.71 [95% CI = 0.5 to 1.0]). Subgroup analysis demonstrated that there was no difference between overall survival in the arms for T1/T2 stage (P = .74, HR = 0.93 [95% CI = 0.59 to 1.4]), whereas there was a difference between the arms for T3/T4 stage (P = .013, HR = 0.51 [95% CI = 0.3 to 0.88]), favoring the CRT arm. The regimen of weekly concurrent CRT is a promising standard treatment strategy for locoregionally advanced nasopharyngeal carcinoma patients. PMID- 15812081 TI - Re: Mesenchymal stem cells: potential precursors for tumor stroma and targeted delivery vehicles for anticancer agents. PMID- 15812082 TI - Re: Continuing outcomes relevant to Evista: breast cancer incidence in postmenopausal osteoporotic women in a randomized trial of raloxifene. PMID- 15812083 TI - Re: Familial clustering of Hodgkin lymphoma and multiple sclerosis. PMID- 15812084 TI - The irreversible damage caused by surreptitious prescribing. PMID- 15812085 TI - Use of prescription psychiatric drugs and religious service attendance. PMID- 15812086 TI - Psychiatric rehabilitation in a Chinese psychiatric hospital. PMID- 15812087 TI - Access to psychiatrists in the public sector and in managed health plans. PMID- 15812088 TI - Implementing a multisource outcome assessment protocol in a state psychiatric hospital: a case study. PMID- 15812089 TI - Bringing grandma home. PMID- 15812091 TI - Using the GAF as a national mental health outcome measure in the Department of Veterans Affairs. AB - OBJECTIVE: Data from the Veterans Health Administration (VHA) were used to evaluate the strengths and weaknesses of the Global Assessment of Functioning (GAF), a single-item mental health status measure, as an outcome measure for large mental health care systems. METHODS: The sample consisted of VHA mental health patients who had at least two GAF scores 45 days apart in 2002 (N=283,754). First, to evaluate the discriminant validity of the GAF change measures, the authors examined the association of these measures with sociodemographic and clinical characteristics. Facility-level risk-adjusted measures of GAF change were then created in three different clinical samples at more than 130 VHA medical centers, adjusting for patients' sociodemographic characteristics and diagnoses. The internal consistency of the scale created by using these items and their consistency across medical centers over time was evaluated. RESULTS: The analysis supported the discriminant validity of the GAF derived measures. As expected, veterans who had a diagnosis of schizophrenia or Alzheimer's disease or who had service-connected disability ratings above 50 percent had lower baseline GAF scores and showed less improvement. The overall GAF performance measure had a high level of internal consistency (a standardized alpha of .85) and was highly consistent across facilities over time. CONCLUSIONS: The results of this study provide preliminary empirical support for cautious use of a GAF-derived scale in monitoring changes in average facility-level outcomes over time. However, because of the potential for gaming of the measures and uncontrolled variation in the scale's administration across facilities, the scale should not be used to compare outcomes across facilities. PMID- 15812092 TI - Continuity of care and clinical outcomes in a national health system. AB - OBJECTIVE: Continuity of care is widely viewed as a key quality indicator for outpatient mental health care. However, few studies have been conducted of the relationship between continuity of care and client outcomes. This study examined the relationship between measures of three aspects of continuity of care (regularity of care, continuity of treatment across organizational boundaries, and intensity of treatment) and the Global Assessment of Functioning (GAF), a single-item mental health status measure, in a national health care system. METHODS: Three analytic samples were derived from a nationwide Department of Veterans Affairs administrative data set: patients with at least one inpatient GAF rating and a later outpatient GAF rating (N=8,334) and two groups who had at least two outpatient GAF ratings, one group that was beginning a new episode of treatment (N=49,946) and a second group in ongoing treatment (N=123,371). Hierarchical linear modeling was used to control for potential site-level autocorrelation and to adjust for differences in diagnostic status, sociodemographic characteristics, baseline GAF score, and the length of time between GAF ratings. RESULTS: Several positive and significant relationships were found for discharged inpatients and new outpatients. However, only a few of these relationships could be confidently said to be clinically meaningful. Specifically, among discharged inpatients, for every additional month in which an outpatient visit occurred over a six-month period, there was a .69 increase in the GAF change score for a total increase of 4.1 points. Among new outpatients the equivalent values were smaller, at .3 and 1.8. In contrast with the findings for discharged inpatients and new outpatients, high intensity of care was negatively associated with GAF change scores for continuing outpatients. CONCLUSIONS: In contrast with several earlier studies, this study showed positive and statistically significant associations between several continuity-of-care measures and client outcomes. These relationships were observed only in transitional treatment situations, that is, after hospital discharge and at the beginning an episode of outpatient care, situations in which continuity of care may be especially important. However, although positive and statistically significant, the magnitude of these effects may not have been clinically meaningful. PMID- 15812093 TI - Reliability of global assessment of functioning ratings made by clinical psychiatric staff. AB - OBJECTIVE: In the Swedish psychiatric care system, systematic follow-up of clinical work with patients is becoming a part of regular service, and a number of care providers are using the Global Assessment of Functioning (GAF) to measure outcomes. This study investigated the reliability of the GAF and analyzed certain factors that affect measurement errors when the scale is used by regular psychiatric staff. METHODS: Eighty-one raters from various psychiatric outpatient clinics rated eight case vignettes. Interrater reliability was assessed by using intraclass correlation coefficients (ICCs), and factors associated with reliability were analyzed by using raters' unique residual values. RESULTS: The results showed that staff who are responsible for assessing first-time patients at outpatient psychiatric clinics and making diagnoses are using the GAF with satisfactory reliability (ICC(1,1)=.81). The factors associated with reliability were raters' subjective attitude toward the GAF and motivation to use the scale and other measurement instruments in psychiatry. CONCLUSIONS: GAF ratings made by an individual rater can be used to measure changes and outcomes at the group level. However, the measurement error is too large for assessment of change for an individual patient, in which case it might be necessary to use several raters. If raters are positively inclined to use rating instruments, measurement errors are minimized and reliability is maximized. PMID- 15812094 TI - Relationship between the Global Assessment of Functioning and other DSM axes in routine clinical work. AB - OBJECTIVES: The aim of this study was to investigate the validity of the Global Assessment of Functioning (GAF) when routinely used in clinical work by focusing on the relations between the GAF, other axes in the DSM system, and some demographic variables conceptually derived on theoretical and clinical grounds. METHODS: A clinical database containing data for 5,538 patients assessed by 181 raters as a part of routine practice in psychiatric outpatient settings in Sweden was used. A hierarchical linear regression model and a variance component model were used to analyze the data. Regression models were also used to determine how the relation between the GAF and axis I depends on the selection of diagnostic groups in the sample. RESULTS: Seventeen percent of the systematic variance in GAF scores was explained by diagnostic differences as defined on DSM-IV axis I, and 5.1 percent was explained by psychosocial and environmental problems as measured on DSM-IV axis IV. Unexpectedly, the site of the investigation explained another 3.6 percent of the variance. CONCLUSIONS: The GAF can be used as a comprehensive measure of global mental health in routine clinical work for assessment and for outcome management. PMID- 15812095 TI - Selection of outcome assessment instruments for inpatients with severe and persistent mental illness. AB - The purpose of this article is to describe a procedure to assist in selecting outcome measures for inpatients treated at a state psychiatric hospital. The procedure combines evidence-based criteria from the literature, instruments shown to be sensitive to change in clinical trials, and the perspectives of a multidisciplinary team of researchers, administrators, providers, and patient advocates. Recent efficacy and effectiveness studies were used to identify recurrently used outcome instruments. A computerized search of more than 30 bibliographic databases, such as PsycINFO, MEDLINE, Social SciSearch, and ERIC, was conducted for articles published between 1990 and 2002. Comparisons of the most frequently used instruments were made on seven criteria proposed as best practice indicators, including sensitivity to change and robust psychometrics. The sample produced 110 measures. Rater-completed instruments were represented more often than patient-completed ones. However, considerable variability across both methods was found on the criteria. The limited resources associated with publicly funded inpatient facilities led to a recommendation to select at least one rater-completed and one patient-completed instrument. PMID- 15812096 TI - Determinants of indicated versus actual level of care in psychiatric emergency services. AB - OBJECTIVE: This study was undertaken to improve understanding of the admission decision process by distinguishing between the clinically indicated level of care and actual level-of-care decisions in emergency psychiatry. METHODS: Clinicians in emergency psychiatric services in Rotterdam, The Netherlands, prospectively rated 720 patients by using the Severity of Psychiatric Illness Scale and collected information on demographic, clinical, and contextual parameters. The clinically indicated level of care and actual level-of-care decisions were studied independently, by using multivariate logistic regression analyses. The decision-making process was divided into three consecutive steps: evaluation of clinically indicated inpatient or outpatient level of care (step 1), voluntary or involuntary admission (step 2), and actual admission of patients for whom voluntary admission was indicated (step 3). RESULTS: Each step was determined by separate factors. Specifically, clinically indicated admission (step 1) was associated with family or friends' desire for admission (odds ratio [OR]=3.7), previous admissions (OR=2.9), symptom severity (OR=2.7), and personality disorder (OR=.4). Involuntary admission (step 2) was associated with lack of motivation (OR=5.7), symptom severity (OR=3.7), time of referral (OR=3.5) and danger to self or others (OR=2.7). Actual voluntary admission (step 3) was associated mainly with bed availability (OR=8.7). The overall percentage of correctly predicted cases was 82 percent for all steps in the decision process. CONCLUSIONS: This study showed that each step in the admission decision process is determined by a unique set of variables and provided evidence that contextual factors influence decision making. Guidelines for voluntary admission and civil commitment need to be based on the results of studies that distinguish between the clinical needs of patients and contextual factors. PMID- 15812097 TI - Use of general medical services among Medicaid patients with severe and persistent mental illness. AB - OBJECTIVE: The aim of this study was to examine patterns of use of general medical services among persons with a severe and persistent mental illness enrolled in Medicaid from 1996 to 1998. METHODS: A total of 669 persons with a severe and persistent mental illness were identified by using statewide clinical criteria. A three-year database of Medicaid claims was developed to examine service use. The main outcome measures were use of outpatient services for a general medical problem, use of dental and vision services, and use of screening tests for women. Service use was examined by primary psychiatric diagnosis (schizophrenic, affective, paranoid, and anxiety disorders), and analyses controlled for the presence of a chronic medical condition, age, race, and sex. RESULTS: This study found high levels of service use for outpatient services but very low levels for primary and preventive services. Although 78 percent of persons with a schizophrenic disorder had an office-based visit during the three year period, all persons with an anxiety disorder had such a visit. Sixty-nine percent of persons with a schizophrenic disorder had at least one emergency department visit, whereas 83 percent of those with an anxiety disorder had such a visit. Dental and vision visits and the use of mammograms and pap tests followed the same pattern; persons with a schizophrenic disorder had fewer visits and had less overall use than the other diagnostic groups. The use patterns across the four groups were significantly different in outpatient service use, dental and vision service use, and screening tests for women. Compared with persons with a schizophrenic disorder, those with an anxiety disorder were more likely to have had an office-based visit and to have received vision services, those with a paranoid disorder were more likely to have used dental services or received a mammogram, and those with an affective disorder were more likely to have had a pap test. CONCLUSION: Although this group of Medicaid patients with severe and persistent mental illness had access to providers, they received an unacceptably low level of preventive care. Use of health services for general medical problems differed somewhat by primary psychiatric illness. PMID- 15812098 TI - Effects on processes and costs of care associated with the addition of an internist to an inpatient psychiatry team. AB - OBJECTIVE: This study examined the effects of collaboration between an internist and psychiatrists on the processes and cost of care among psychiatric inpatients. METHODS: A randomized controlled study was performed on the psychiatric inpatient units of an academic medical center. All inpatients who were admitted to the units between March 2001 and January 2002 were asked to participate in the study. Patients in the intervention group met with an internist who participated in their care by communicating with the patients' primary care physicians, assessing needs, updating appropriate health maintenance services, managing chronic and acute medical problems, and attending hospital work rounds. Patients in the usual care group received usual care. The processes of care were measured by examining 17 categories that involved assessment of needs, coordination of care, and completion of health maintenance services. Resource use was measured by examining hospital charges and length of stay. RESULTS: A total of 139 adults participated in the study: 55 were in the intervention group and 84 were in the usual-care group. Patients in the intervention group showed significant improvement in 12 of the 17 categories of the processes of care. Overall, no differences were found in hospital charges or length of stay. CONCLUSIONS: Adding an internist to an inpatient psychiatric team is an effective way of improving care for this traditionally underserved population without increasing cost. PMID- 15812099 TI - Depression and comorbid pain as predictors of disability, employment, insurance status, and health care costs. AB - OBJECTIVE: Individuals with depression commonly experience pain with unclear pathology. This study examined depression and comorbid pain and associated outcomes over six years in a nationally representative cohort of older Americans. METHODS: The Health and Retirement Study began in 1992 and follows 9,825 individuals between the ages of 50 and 61 years. The study reported here used data beginning in 1994 to contrast individuals with depression and those with depression plus comorbid pain. Regression models adjusted for differences in sociodemographic characteristics and health status. RESULTS: Baseline (1994) data were available for 8,280 participants. At baseline, 65.2 percent reported that they did not have pain or depression, 8.1 percent had depression alone, 15.5 percent had mild or moderate pain alone, 2 percent had severe pain alone, 6.6 percent had depression plus mild or moderate pain, and 2.6 percent had depression plus severe pain. Compared with the group with no pain or depression, all the groups with depression, pain, or both had greater decrements in outcomes. Groups tended to have greater decrements in outcomes as levels of pain increased. Overall, no statistically significant differences were found between the groups with depression plus pain and their corresponding groups with pain alone. Two to six years after baseline, compared with participants with depression alone, those with severe pain or depression plus severe pain were more likely to experience new functional limitations and to have higher total health care expenditures. Compared with participants with depression alone, participants with depression plus severe pain were also more likely to lose employment and private health insurance. CONCLUSIONS: Relative to depression alone, depression plus pain and pain alone (particularly severe pain) were associated with significant functional limitations and economic burdens. PMID- 15812100 TI - Health service use among persons with comorbid bipolar and substance use disorders. AB - OBJECTIVE: This study tested the hypothesis that patients with comorbid bipolar and substance use disorders use health services to a greater extent than patients with either bipolar or substance use disorder alone. METHODS: A retrospective chart review was conducted among patients who used health services at the Ralph H. Johnson Department of Veterans Affairs medical center in Charleston, South Carolina, and had bipolar disorder alone, substance use disorder alone, and comorbid bipolar and substance use disorders. Patients with a psychiatric admission between 1999 and 2003 were included in the study. Information was collected on the use of health services one year before and including the index admission. RESULTS: The records of 106 eligible patients were examined for this study: 18 had bipolar disorder alone, 39 had substance use disorder alone, and 49 had both bipolar and substance use disorders. Compared with the other two groups, the group with comorbid bipolar and substance use disorders was significantly more likely to be suicidal. Compared with the group with bipolar disorder alone, the group with comorbid disorders had significantly fewer outpatient psychiatric visits and tended to have shorter psychiatric hospitalizations. Among patients with an alcohol use disorder, those who also had bipolar disorder were significantly less likely than those with an alcohol use disorder alone to have had an alcohol-related seizure. Patients with comorbid bipolar and substance use disorders were significantly less likely than those with substance use disorder alone to be referred for intensive substance abuse treatment, even though both groups were equally likely to enter and complete treatment when they were referred. CONCLUSIONS: Despite significant functional impairment among patients with comorbid bipolar and substance use disorders, they had significantly fewer psychiatric outpatient visits than those with bipolar disorder alone and were referred for intensive substance abuse treatment significantly less often than those with substance use disorder alone. PMID- 15812101 TI - Surreptitious prescribing in psychiatric practice. PMID- 15812102 TI - A comparison of the Aberrant Behavior Checklist and the GAF among adults with mental retardation and mental illness. AB - Psychiatric assessment among individuals with a diagnosis of both mental retardation and mental illness presents a clinical challenge. This retrospective study compared two rating scales--the Aberrant Behavior Checklist (ABC) and the Global Assessment of Functioning (GAF)--to determine the scales' utility in a partial hospital setting. Although ABC and GAF ratings were weakly correlated, the ABC revealed symptom patterns consistent with recognizable features of psychiatric syndromes and differential improvement in symptoms within and between diagnostic subgroups. The ABC provided a more useful measure of treatment response than the GAF in this patient population. PMID- 15812103 TI - A pharmacy-based coaching program to improve adherence to antidepressant treatment among primary care patients. AB - The effects on adherence and depressive symptoms of a community pharmacy-based coaching program, including a take-home videotape, were evaluated in a randomized controlled trial in the Netherlands. A total of 147 depressed primary care patients who had a new antidepressant prescription were included in the study. Adherence was measured with an electronic pill container and was also derived from pharmacy medication records; the latter method was associated with an overestimation of adherence of only 5 percent. Intention-to-treat analyses showed no intervention effect on adherence (73 percent compared with 76 percent), whereas analyses of patients who received the intervention (per protocol) showed improved adherence (73 percent compared with 90 percent). Neither analysis showed effects on depressive symptoms. PMID- 15812104 TI - ECT then and now. PMID- 15812105 TI - Weight loss and anorexia with quetiapine. PMID- 15812106 TI - Use of coercive measures in a psychiatric intensive care unit in Slovenia. PMID- 15812107 TI - Visual memory and visual perception recruit common neural substrates. AB - This human neuroimaging review aims to determine the degree to which visual memory evokes activity in neural regions that have been associated with visual perception. A visual perception framework is proposed to identify cortical regions associated with modality-specific processing (i.e., visual, auditory, motor, or olfactory), visual domain-specific processing (i.e., "what" versus "where," or face versus visual context), and visual feature-specific processing (i.e., color, motion, or spatial location). Independent assessments of visual item memory studies and visual working memory studies revealed activity in the appropriate cortical regions associated with each of the three levels of visual perception processing. These results provide compelling evidence that visual memory and visual perception are associated with common neural substrates. Furthermore, as with visual perception, they support the view that visual memory is a constructive process, in which features or components from disparate cortical regions bind together to form a coherent whole. PMID- 15812108 TI - Galanin and cognition. AB - Since the neuropeptide galanin's discovery in 1983, information has accumulated that implicates it in a wide range of functions, including pain sensation, stress responses, appetite regulation, and learning and memory. This article reviews the evidence for specific functions of galanin in cognitive processes. Consistencies as well as gaps in the literature are organized around basic questions of methodology and theory. This review shows that although regularities are evident in the observed behavioral effects of galanin across several methods for measuring learning and memory, generalization from these findings is tempered with concerns about confounds and a restricted range of testing conditions. Furthermore, it is revealed that many noncognitive behavioral constructs that are relevant for assessing potential roles for galanin in cognition have not been thoroughly examined. The review concludes by laying out how future theory and experimental work can overcome these concerns and confidently define the nature of the association of galanin with particular cognitive constructs. PMID- 15812109 TI - The role of attentional bias in substance abuse. AB - There has been much recent interest in the idea that drug users show biased attention toward drug-related events. Because drug stimuli produce conditioned responses that may motivate drug taking, biased attention toward these cues may play an important role in drug use and relapse following treatment. The performance of drug users on the Stroop task and visual dot-probe task has been interpreted as demonstrating attentional bias toward drug cues specific to an individual's drug use history. However, studies often fail to include necessary control groups or comparison stimuli, thereby making it difficult to definitively conclude that reported results reflect a specific attentional response to personally relevant drug events. Although promising, these initial studies need to be followed up with better controlled demonstrations of attentional bias and with studies linking bias levels to other measures of drug taking. PMID- 15812110 TI - Measuring impulsivity and modeling its association with cigarette smoking. AB - Two types of behavioral measure are primarily used to examine impulsivity in humans and animals: Go/No-go tasks to assess inhibition and relative preference tasks to assess delay aversion. Several examples of each type of task are described so that common cognitive processes and variables affecting performance can be identified. Data suggest that smokers are more impulsive on each of these impulsivity measures than nonsmokers. Several models can be proposed to account for this group difference: (a) the differences predate and, possibly, are causally related to the initiation of cigarette smoking; (b) higher levels of impulsivity are associated with continued smoking, either through an association with heightened positive subjective effects of nicotine or heightened negative effects of nicotine abstinence (withdrawal); (c) nicotine causes neuroadaptations that result in elevated impulsivity in smokers. Studies relating to each of these models are reviewed, and it is concluded that all three models may contribute to the observed higher levels of impulsivity in smokers. However, pertinent studies are limited and additional systematic research is needed. PMID- 15812111 TI - Outcome of parapneumonic empyema. AB - OBJECTIVE: Empyema thoracis is known to have variable age group affection, causative agents and controversy regarding primary mode of management. To look into current demography, bacteriology and treatment outcome. METHODS: Prospective study made on admitted cases of parapneumonic empyema from July 2001 to June 2003. All cases were treated with chest tube drainage, parenteral antibiotics or thoracotomy in multiloculated or non-improving cases. RESULTS: 0.8% (C.I. 0.6 1.0) of total pediatric admission had empyema, who were more likely to be females (P< 0.05), under-weight (P< 0.05) compared to children admitted for other reasons. Staphylococcus aureus is still the commonest isolate (13.2%). All cases received antibiotics prior to hospitalisation. Majority of cases (90.5%) could be successfully managed with antibiotics and chest tube drainage alone. 9.4% cases needed thoracotomy. 5.8% cases needed salvage thoracotomy following non improvement with chest tube drainage. Fever remission time and duration of hospital stay were comparable in both groups. Thoracotomy cases required antibiotics for shorter period (P=0.04). Two cases died due to reasons other than mode of management. Radiological and lung function recovery was excellent in most of the cases. CONCLUSION: Chest tube drainage is a safe, efficacious primary method of empyema management. PMID- 15812112 TI - Acute scrotum -- etiology and management. AB - OBJECTIVE: Acute scrotal pain is a common urological emergency. Urgent exploration is the standard means of management, since no investigation can confidently exclude torsion of testis from the differential diagnosis. METHODS: A review of all boys presenting with acute scrotal pain who underwent emergency scrotal exploration between January 1983 and March 2003 was performed. RESULTS: 195 boys were included in this study. They were divided into 3 groups: group 1-73 with epididymo-orchitis, group 2-63 with torsion of testicular appendages and group 3- 57 with spermatic cord torsion. The patients in group 2 were older than group 1, also patients in group 1 were older than group 3. During neonatal period the most common pathology was spermatic cord torsion, whereas in prepubertal period torsion of appendages was more common. In all boys, mean duration of pain at presentation was 2.11 days. Epididymo-orchitis was diagnosed in 37% of patients, with torsion of the appendages being the next most common entity. Testicular torsion was diagnosed in 29% of patients. In the group with testicular torsion salvage rate of testis was 37% because of late admission. The perioperative morbidity and mortality was not seen in any of the groups. CONCLUSION: We believe that any boy with acute scrotal pain and any suspicion of testicular torsion in physical examination must be applied routine surgical exploration. PMID- 15812113 TI - Relationship between serum bilirubin and coagulation test results in 1-month-old infants. AB - OBJECTIVE: Although the connection between cholestasis and conjugated hyperbilirubinemia is well known, mild hepatic dysfunction or cholestasis may also be associated with unconjugated hyperbilirubinemia in some infants with prolonged jaundice. The aim of this study was to investigate the relationship between serum bilirubin levels and alanine aminotransferase levels, aspartate aminotransferase levels, prothrombin time, activated partial thromboplastin time, and international normalization ratio findings in a group of infants. METHODS: The study included 77 healthy, term, breast-fed infants with jaundice and 56 age matched, healthy, term, non-jaundiced controls. The 133 babies were divided into three subgroups according to their total bilirubin levels [group I (controls) < 50 micromol/L, group II = 50-100 micromol/L, and group III > 100 micromol/L, and the findings for the noted parameters were compared]. RESULTS: The mean conjugated bilirubin level was significantly higher, and the mean activated partial thromboplastin time significantly longer in group III than in group I. A significant positive correlation was found between bilirubin levels and PT and APTT results. CONCLUSION: Clinical vitamin K deficiency appeared unlikely to develop in this group of infants with prolonged unconjugated hyperbilirubinemia. However, a significant positive correlation between bilirubin levels and PT and APTT suggest that a higher bilirubin load to the liver may cause some degree of vitamin K deficiency due to mild cholestasis. The importance of this finding, and the possible benefits of vitamin K supplementation in 1-month-old breast-fed infants with bilirubin levels higher than 100 micromol/L require further investigation. PMID- 15812114 TI - Breastfeeding and pacifier use in Brazil. AB - OBJECTIVE: To determine the relationship between pacifier use and the duration of exclusive breast-feeding in the first six months of age, among poor children with unfavourable birth weight, from an underdeveloped region in Brazil. METHOD: Prospective cohort study with infants followed from birth to 6 months of age. Healthy children born with unfavourable birth weight (< 3,000), being exclusively breastfed, were selected from 8 maternity hospitals in the city of Fortaleza (Brazil) between November 1996 and April 1997. Two main outcome measures were used: (i) time to stop exclusive breast-feeding at the 1st and (ii) at the 6th month of life. Main exposures were pacifier use at 1st and 6th month of age. Data were collected at maternity hospitals and during home interviews, using structured questionnaires, by trained data collectors unaware of the study aims, and analyzed using survival analysis and the Cox Proportional Hazard Model. RESULTS: 500 children were enrolled and 13% were lost to follow up at the 1st month. Most of the families had a monthly income less than five times the minimum wage. One third of the mothers were adolescents, one fifth were working outside the home by the 6th month and most attended prenatal care visits. Approximately 60% of the children were using pacifiers by the 1st month. The average number of days for exclusive breast-feeding for pacifier use by the 6th month was 125.3 compared to 87.0 among non-users (p=0.0001). Children using pacifiers were 1,9 more likely to have stopped exclusive breastfeeding by the 6th month compared to non-users, even after controlling potential confounders. CONCLUSION: Pacifier use was associated with the early termination of breast-feeding in Brazil, among poor children with unfavourable birth weight, living in an underdeveloped area. As a possible marker of early weaning, pacifier use can help health workers identify those mothers in need of extended counselling to reinforce breast-feeding practices. PMID- 15812115 TI - Tuberculin reactivity in tuberculous meningitis. AB - OBJECTIVE: To study tuberculin reactivity in childhood tuberculous meningitis both in clinical and histopathological (HP) context. METHODOLOGY: Children with tuberculous meningitis (TBM) were given tuberculin test by Mantoux technique, which was read at the end of 72 hours after the placement of skin test. Histopathological examination of the punch biopsy specimen of the tuberculin test site was performed and histopathological grading of the tuberculin reaction was compared with clinical reaction and clinical parameters. RESULTS: Of the 50 children studied, 68% of them were malnourished and 42% had BCG scar. Tuberculin test was positive in 22 (44%) cases. Spearman analysis showed negative correlation between stage of TBM and the size of tuberculin reaction. BCG status did not affect the size of tuberculin reaction. Histopathological grade of the tuberculin reaction was found to be directly proportional to the size of the tuberculin reaction and it was not affected by the stage of TBM. CONCLUSION: Tuberculin positivity is low in TBM irrespective of the nutritional status. At least some degree of inflammatory reaction can be seen at the site of tuberculin administration. In tuberculin negative cases, varying grades of cellular response in the absence of clinical induration can be seen in histopathology. PMID- 15812116 TI - Reactions of Indian adolescents to the 9/11 terrorist attacks. AB - OBJECTIVE: There is information about the impact of disasters and trauma on children, but little is known about the effects of terrorism particularly in India. (i) To assess the knowledge of the 9/11 terrorist attacks to the school going adolescents of India who were miles away from the actual incident. (ii) To compare the reactions to this event among the boys and girls. METHODS: The study used a survey design with a self-report questionnaire administered to 406 students in 6 schools of standards 9-12. The questionnaire was administered within 3 weeks of occurrence of this event. The mean age of the subjects was 16.34 years (SD= 1.22; range= 13-20) and 44.1% were boys. RESULTS: All the students were aware about this event. Awareness that the twin towers were hit was in 81.06 but only 51.94% knew that Pentagon was also hit. All the children knew who the prime suspect was although only 12.62% were aware about the country to which he belonged. The source of knowledge of the events was the television in 74.7% of the adolescents and 17.95% of them viewed foreign news channels additionally to the Indian channels to gather details about the event. Newspaper, radio and internet were the sources of information in 44.17%, 3.4% and 3.5% children respectively. Of the adolescents who gathered information from the television, 84.7% agreed that there had been an increase in their TV viewing time since the event and it was more than one hour per day in 47.5% of them. None of the students supported the terrorist attacks. The number of students with negative stressors was significantly more than the ones who were unaffected (p 0.0001). The girls were significantly more affected than the boys and while the former expressed anger the latter were more fearful and sad. (p 0.05) The idea of USA going for war against Afghanistan was supported by 69.4% and one third of them believed that such an event might adversely affect India. The adolescents who had witnessed the events on television were more fearful and shocked than the ones who read about the event in the print media (p 0.05). CONCLUSION: This study emphasizes the adverse reactions in the minds of adolescents in India to terrorist events even though they did not directly witness the events of September 11,2001. The role of media exposure in causing stress is also revealed. Pediatricians should be aware of the adverse effects of terrorism in the minds of the children and should be able to identify and help those who are in need. PMID- 15812117 TI - Spina bifida occulta in functional enuresis. AB - OBJECTIVE: To study incidence of spina bifida occulta in nocturnal enuresis cases and to compare outcome of enuresis with spina bifida occulta and enuresis without spina bifida occulta. METHODS: Patients with enuresis divided into two groups based on X-ray lumbosacral spine. Outcome of these patients were compared on behavioral therapy. RESULTS: Spina bifida occulta was detected in 18 out of total 48 patients. Levels of spina bifida were L5 vertebrae in 3,L5-S1 in 5,S1 in 8 and S1-S2 in 2 patients. Outcome was evaluated in 42 patients who were followed up for more than 6 months. There was no significant difference between both the groups. CONCLUSION: Spina bifida occulta is a common finding in enuresis. Outcome of patients with spina bifida occulta is not different than the patients without spina bifida occulta. PMID- 15812118 TI - Autism -- experiences in a tertiary care hospital. AB - Pervasive developmental disorders (PDD) or Autistic Spectrum Disorders (ASD) include Autistic Disorder (commonest), Asperger's syndrome, Childhood Disintegrative Disorders, Rett's syndrome and PDD-NOS (not otherwise specified). OBJECTIVE: Autism is an important cause of social disability and reported more often from the developed world than from the developing countries. The present study was aimed to establish the diagnosis of autism amongst children with derangements of language, communication and behavior; ascertain and treat the co morbidities; identify underlying cause and create a sensitivity and awareness among various health care professionals. METHODS: Sixty-two of the seventy-five referred patients fulfilled the DSM-IV (Diagnostic and Statistical Manual of Mental Disorder) criteria for autism. Evaluation included a detailed history, clinical examination, IQ assessment, Connor's scoring for hyperactivity and Fragile-X screening. Management of co-morbidities was done. A follow up of these patients was done. Parents' assessment of the child was also done. A registry for autistic children was established at the Department of Pediatrics with other major institutions of Delhi. RESULTS: The male:female ratio was 8:1 and missed diagnosis was common. Professional awareness is merited. Behavioral modification by early intervention and stimulation improved the core symptoms of autism. Important co-morbidities included mental retardation (95%), hyperactivity (53%) and seizures (10%) cases. Control of co-morbidities in these children facilitated child's periodic assessment and implementation of intervention programmes. In the registry initiated 62 patients were enrolled at AIIMS and 6 were identified from other hospitals. CONCLUSION: Autism does occur in Indian children too. Diagnosis is often missed. Capacity building among health professionals by a more structured teaching of developmental disabilities in the medical curriculum is required. The need to attend to co-morbidities and associated symptoms was clear. The initiation of the registry and beginning of networking was important. PMID- 15812119 TI - Comparative efficacy and safety evaluation of cefaclor vs amoxycillin + clavulanate in children with Acute Otitis Media (AOM). AB - Acute Otitis Media (AOM) is the most frequent respiratory tract infection of infancy and childhood that is treated with antimicrobial agents. The most common causative pathogens include Streptococcus pneumoniae, Hemophilus influenzae and Moxarella catarrhalis, and therefore antibacterial management should target against these isolates. Cefaclor, a congener of cephalexin monohydrate, is a semisynthetic cephalosporin antibiotic. It is an orally active cephalosporin which has demonstrated activity against a wide range of organisms in vitro. Present study is designed as a multicentric prospective trial to study and compare the efficacy and safety of cefaclor versus amoxicillin + clav in children with acute otitis media. One hundred and sixty seven patients were evaluated for efficacy endpoints in the cefaclor arm comprised of 104 males and 63 females with a mean age of 5.74+/-2.80 years and 185 patients in the amoxy-clav group comprised of 118 males and 67 females with a mean age of 4.93+/-2.92 years. Both cefaclor and amoxy-clav caused a significant improvement in all the signs and symptoms after a 10-day treatment period. However, between-the-group comparisons showed that the reduction in most of the symptoms was significantly more in cefaclor arm as compared to amoxicillin-clav arm. The clinical success (clinical cure + improvement) at the end of therapy was significantly more in cefaclor arm: 98% with cefaclor versus 85% with amoxicillin + clav, p<0.05 Table 3. Failure cases were prescribed other antibiotics according to the culture sensitivity reports, as rescue medication. Bacterial eradication rates were largely consistent with clinical responses. Bacteriological eradication was seen in 95% of patients in cefaclor group and 78% of patients in amoxicillin + clav group. In conclusion, cefaclor is a well tolerated and effective antibacterial option for acute otitis media in children and it is superior to the combination of amoxicillin + clav in efficacy and tolerability in acute AOM. Moreover, its expanded spectrum of activity, ability to achieve adequate concentrations in tissues, suitability for twice-daily dosing, and proven tolerability suggest that it is a good alternative to agents traditionally used in acute otitis media. PMID- 15812120 TI - Essential fatty acids, DHA and human brain. AB - Essential fatty acids cannot be synthesized in the body but they are required for maintenance of optimal health. There are two classes of polyunsaturated fatty acids (PUFAs)--omega-6 and omega-3. The parent omega-6 fatty acid, linoleic acid (LA) is desaturated in the body to form arachidonic acid while parent omega-3 fatty acid alpha-linolenic acid (ALA) is desaturated by microsomal enzyme system through a series of metabolic steps to form eicosapentaenoic acid (EPA) and decosahexaenoic acid (DHA). But there is a limited metabolic capability during early life to metabolize PUFAs to more active long-chain fatty acids. There is a critical role of EFAs and their metabolic products for maintenance of structural and functional integrity of central nervous system and retina. Most of the brain growth is completed by 5-6 years of age. At birth brain weight is 70% of an adult, 15% brain growth occurs during infancy and remaining brain growth is completed during preschool years. DHA is the predominant structural fatty acid in the central nervous system and retina and its availability is crucial for brain development. It is recommended that the pregnant and nursing woman should take at least 2.6 g of omega-3 fatty acids and 100-300 mg of DHA daily to look after the needs of her fetus and suckling infant. The follow-up studies have shown that infants of mothers supplemented with EFAs and DHA had higher mental processing scores, psychomotor development, eye-hand coordination and stereo acuity at 4 years of age. Intake of EFAs and DHA during preschool years may also have a beneficial role in the prevention of attention deficit hyperactivity disorder (ADHD) and enhancing learning capability and academic performance. PMID- 15812121 TI - Sodium valproate -- induced skeletal myopathy. AB - The authors report a case of skeletal myopathy in a four-year-old boy on long term sodium valproate therapy for secondary epilepsy due to neurocysticercosis. He presented with clinical features of limb girdle weakness. EMG revealed features of myopathy. Carnitine deficiency due to sodium valproate was suspected and plasma carnitine levels were found to be low. Sodium valproate was withdrawn. L-carnitine supplementation resulted in marked clinical recovery as well as rise in plasma carnitine levels. PMID- 15812122 TI - Scimitar syndrome. AB - Scimitar syndrome, a rare anomaly of partial pulmonary venous drainage into the hepatic portion of the inferior vena cava, detected in a ten-day-old neonate who presented clinically with respiratory distress and diagnosed on ultrasound of the thorax is being reported here. PMID- 15812123 TI - Scimitar syndrome. AB - Scimitar syndrome is a relatively uncommon constellation of cardio-pulmonary anomalies, its typical feature being partial anomalous pulmonary venous connection. It can present in the neonatal period as well as later in life. We present the case of a girl diagnosed in the newborn period, along with a brief review of literature. PMID- 15812124 TI - Open indirect cardiac massage in neonate. AB - Open, direct cardiac massage (OCM), as a technique of resuscitation has not been very inspiring due to its various limitations. We report a case of resuscitation where open 'indirect' cardiac massage was used to resuscitate a neonate successfully. A 24 hr old neonate with multiple gastrointestinal anomalies presented for emergency laparotomy. Intraoperatively she suffered a cardiac arrest. External chest compressions (ECC) failed to produce detectable circulation and surgeons were asked to initiate direct cardiac compressions. They could palpate and successfully compress the heart without the need to split the diaphragm. This 'infra-diaphragmatic' approach resulted in adequate circulation and subsequent uncomplicated recovery. PMID- 15812125 TI - Rapidly progressive IgA nephropathy. AB - A 14-year-old boy presented with macroscopic hematuria and a rapid deterioration in renal function. Percutaneous renal biopsy demonstrated severe crescentic IgA nephropathy (IgAN) with extensive (88%) glomerular crescent formation. After started intravenous administration of high-dose pulse methylprednisolone, severe nausea and general malaise accompanied by a rapid increase in Blood Urea Nitrogen (BUN) and serum creatinine levels appeared, however, the renal function ameliorated rapidly and fully recovered by following oral administration of corticosteroid. The clinical presentation of our case seems to be very remarkable compared to previously reported cases of rapidly progressive IgAN. PMID- 15812126 TI - Pantothenate kinase associated neurodegeneration (Hallervorden-Spatz syndrome). AB - Hallervorden-Spatz syndrome is a rare autosomal recessive hereditary condition characterized by early onset of progressive movement alteration that include dystonia, rigidity and choreoathetosis usually associated with pyramidal signs and mental deterioration. We report two sisters where diagnosis was missed till MRI showed classic imaging findings. Mutation analysis in one, revealed homozygous mutations in the PANK 2 gene. The need for clinical recognition of this entity and differentiation of this form from other static and progressive neurological illnesses is emphasized. PMID- 15812127 TI - Obstructive uropathy as cause of rickets: often a forgotten entity. PMID- 15812129 TI - Traumatic neuritis and AFP surveillance. PMID- 15812130 TI - Sodium stibogluconate and polymorphic ventricular tachycardia. AB - Numerous antimicrobials including pentavalent antimonials are implicated in causing prolong QT-interval and ventricular tachycardia. Torsades de pointes is rarely documented with use of sodium stibogluconate. Here is described a 12-yr old girl with visceral leishmaniasis, who developed syncopal attacks, prolong QT interval, polymorphic ventricular tachycardia and torsades de pointes after completing a course of Stibogluconate (20 mg/kg/day for 30 days). Prolong lidocaine infusion and cardioversion were life saving. PMID- 15812131 TI - An extradural and subdural hematoma in a neonate. AB - Traumatic brain injury following birth is common in newborn but significant intracranial haematoma following birth injury is not that usual. Even busy pediatric trauma center have about only 1 to 3 % of admission that require neurosurgical care. Extradural haematoma (EDH) associated with intracerebral and subdural haematoma (SDH) is even more rare in newborn. If this is not detected and treated in time, the outcome may be fatal. A case of EDH with subdural and intracerebral haematoma in a 3 days old neonate is presented. Etiology and problems in diagnosis and management are discussed. PMID- 15812132 TI - Jeune thoracic dystrophy with right sided diaphragmatic hernia. AB - Jeune thoracic dystrophy is a rare autosomal recessive chondrodysplasia, first described by Jeune et al in 1955. Early death is usually the consequence of asphyxia with or without pneumonia. Here is reported a newborn with Jeune thoracic dystrophy and a right-sided diaphragmatic hernia. PMID- 15812133 TI - Fetomaternal alloimmune thrombocytopenia presenting as intracerebral bleeding in utero. AB - Feto-maternal alloimmune thrombocytopenia is a serious fetal disorder resulting from platelet antigen incompatibility between the mother and the fetus. Intracranial bleeding is the most serious complication of alloimmune thrombocytopenia and can result in severe disability and death in utero. The authors report a case of intracerebral hemorrhage in utero resulting from alloimmune thrombocytopenia. PMID- 15812134 TI - [Neural mechanisms of recognition of facial expression in the amygdala]. PMID- 15812135 TI - [Behavioral approaches for analyzing emotionality in mutant mice]. PMID- 15812136 TI - [Emotional stress and functional development of the rat brain]. PMID- 15812137 TI - [Neural basis of emotion and feeling from a neuropsychological perspective]. PMID- 15812138 TI - [Development of neuroimaging research on human emotion]. PMID- 15812139 TI - [Methods for evaluating an analgesic effect on pain in awake behaving monkeys]. PMID- 15812140 TI - [Actin polymerization kinetics in living cells disclosed by single-molecule speckle microscopy]. PMID- 15812141 TI - Chicken RFamide-related peptide (GnIH) and two distinct receptor subtypes: identification, molecular characterization, and evolutionary considerations. AB - RFamide-related peptides (RFRPs) regulate the release of various pituitary hormones in vertebrates. It is completely unknown how the functions of RFRPs vary among animal classes and whether vertebrate RFRPs are orthologous to each other and belong to the same peptide family. This report concerns identification of avian RFRP (gonadotropin-inhibitory hormone, GnIH) from the chicken. Chromosome wide synteny conservation demonstrated the orthologous relationships among vertebrate RFRPs. The consensus motif for RFRP was modified to Pro-Xaa-Arg-Phe NH2. We also describe the first identification of two distinct types of receptors for non-mammalian RFRP (RFRPR and NPFFR) from the chicken. Amino acid comparison revealed substantial differences in both termini of receptors among classes of vertebrates. The 5'-flanking regions of chicken RFRPR and NPFFR suggested their expressions in the pituitary gland, and this was confirmed by the RT-PCR analysis. Localizations of both chicken RFRP and its receptors were distinct from those of mammals. These results indicated that avian RFRP, unlike the mammalian one, directly acts on the pituitary gland via receptors to regulate gonadotropin release. It was also suggested that functional differences, especially between avian and mammalian RFRPs, are substantially due to divergences in the structures and expression sites of their receptors. PMID- 15812142 TI - Usage of putative chicken U6 promoters for vector-based RNA interference. AB - Gene silencing with short interfering RNA (siRNA) expression vectors is a powerful method for the analysis of gene functions. For the expression of siRNA in mammalian cells, mammalian U6 small nuclear RNA (snRNA) promoters are widely used. However, the mammalian U6 promoter might not function well in other species. In this study, we cloned four putative chicken U6 promoters by PCR and analyzed their functions. First, we screened the chicken genomic database using the human U6 snRNA gene and identified four candidate sequences. The sequences contained some control elements in their promoter regions, but as we could not rule out that they were pseudogenes, we amplified these sequences and used them as promoters for short hairpin RNA (shRNA) expression. Using the firefly luciferase (Luc) gene as a target, transient expression assays were performed with chicken ovary-derived cells. All four putative chicken U6 promoters exhibited suppressive activity toward Luc, and so could act as a promoter for expression of the snRNA gene in the chicken genome. The promoter activity was not as strong as that of a commercially available siRNA expression vector. This probably reflects artificial sequences between the promoters and synthetic DNA encoding shRNA. PMID- 15812143 TI - Effects of maturation conditions on spindle morphology in porcine MII oocytes. AB - Incomplete cytoplasmic maturation of in vitro matured (IVM) oocytes has been known to cause microtubule and microfilament alterations, which may result in abnormal pronuclear formation and failed embryonic development. We examined the influences of maturation conditions on meiotic spindle morphology at metaphase of meiosis II (MII) in porcine oocytes. Porcine oocytes were matured under various conditions, i.e., in vitro or in vivo, with different amounts of cumulus cells, with or without hormonal supplements, and with various exposure durations to the hormones, to examine the effects on spindle morphology in MII oocytes by immunofluorescence under confocal laser microscopy. Interpolar spindle length (microm) and spindle area (microm2) were compared among these maturation conditions. The spindle length was significantly shorter in IVM oocytes compared to those matured in vivo. Oocytes collected from cumulus oocyte complexes (COCs), which were poor in cumulus cells, showed smaller spindle areas than those from cumulus-rich COCs. The spindle length and area were both significantly reduced in oocytes grown without hormonal supplements. When oocytes were grown with hormonal supplements for either 6 or 22 hours for the first half of culture, there was no difference in the spindle morphology between these oocytes. These results suggested that maturation conditions significantly influence morphogenesis of MII spindles in porcine oocytes. Oocytes matured in poor conditions were more likely to have a shorter spindle length (long axis) and smaller spindle areas. PMID- 15812144 TI - Cloning of the elk TSH beta-subunit cDNA and seasonal expression of the pituitary glycoprotein hormone genes. AB - We report the elk (Cervus elaphus) thyroid stimulating hormone (TSH) beta-subunit cDNA cloning, nucleotide and deduced amino acid sequences. The TSH beta-subunit cDNA was obtained by RT-PCR of polyadenylated pituitary RNA. The deduced elk TSH beta-subunit peptide chain shares between 93 to 99% sequence similarities with the reported TSH beta-subunit of a sub-set of related species. The TSH beta subunit gene is expressed in the elk pituitary gland as a mature transcript of approximately 600 bases, which corresponds to the size of the mRNA expressed in the sheep pituitary gland. Seasonal expression of the pituitary gonadotropin genes was investigated by Northern blot analyses. Samples of elk pituitary glands collected during the breeding season showed elevated steady state levels of common alpha-subunit and FSH and LH beta-subunit gene expression, consistent with the seasonal reproductive cycling of this species. Samples collected before the breeding season demonstrated decreased expression of the gonadotropin genes. TSH, which is not directly tied to reproduction, had similar levels of expression, regardless of the animal's reproductive status. PMID- 15812145 TI - Apoptotic response to albumin overload: proximal vs. distal/collecting tubule cells. AB - End-stage renal disease due to proteinuric states has a great impact on the quality of life by necessitating renal replacement therapy. Understanding the pathophysiologic consequences of proteinuria is crucial in order to develop treatment strategies to halt the progression. We have previously reported that cultured porcine proximal tubule cells respond to albumin overload by undergoing apoptosis. In this study, we investigated the differential apoptotic response to albumin in HKC-8 (proximal tubule) and MDCK (collecting/distal tubule) cells under high concentrations of albumin simulating the nephrotic milieu. Our results are consistent with marked cytotoxicity and apoptosis within 24 h of albumin incubation in HKC-8 cells that was closely related to the fatty acid content of the albumin. In contrast, in MDCK cells, albumin stimulated cell turnover by stimulating proliferation and late onset apoptosis regardless of the fatty acid content. Another important result of this study is the direct demonstration of albumin uptake by MDCK cells mediated by endocytosis via clathrin-coated pits. A comparison of albumin uptake between proximal and distal/collecting tubule cells revealed faster uptake in proximal tubule cells within 15 min but almost 100% albumin uptake of both cell types in 1 h. In summary, our findings demonstrate that both proximal and distal nephron segments are affected in proteinuric states, but the degrees of susceptibility to albumin and associated lipid moieties are distinct in the different nephron segments. PMID- 15812146 TI - Modification of collagen matrices for enhancing angiogenesis. AB - The vascularization of engineered tissues in many cases does not keep up with the ingrowth of cells. Nutrient and oxygen supply are not sufficient, which ultimately leads to the death of the invading cells. The enhancement of the angiogenic capabilities of engineered tissues therefore represents a major challenge in the field of tissue engineering. The immobilization of angiogenic growth factors may be useful for enhancing angiogenesis. The most potent angiogenic growth factor specific to endothelial cells, vascular endothelial growth factor (VEGF), occurs in several splice variants. The variant with 165 amino acids both has a high angiogenic activity and a high affinity for heparin. We therefore incorporated heparin molecules into collagen matrices by covalently cross-linking them to amino functions on the collagen. Physical binding of VEGF to the heparin may then prevent a rapid clearance from the implant, while the release rate may become coupled to the degradation of the collagen matrix. The modified matrices were characterized by determination of the extent of the heparin immobilization, the in vitro degradation rate by collagenase. For testing the angiogenic properties, non-modified and heparinized collagen specimens were- either loaded with VEGF or non-loaded--subcutaneously implanted on the back of rats. Specimens were explanted after varying periods of implantation, the dry weights and the hemoglobin contents, as well as immunostained histological sections were evaluated: heparinized collagen matrices loaded with VEGF are vascularized to a substantially higher extent as compared to non-modified matrices. PMID- 15812147 TI - Geometric and elastic properties of in vivo human Achilles tendon in young adults. AB - The purpose of this study was to clarify whether the major determinant of the extendibility of the Achilles tendon in young adults was the geometric properties of the tendon. The subjects were 38 healthy young adults (26 male, 12 female; 26 +/- 5 years). The subjects developed maximum voluntary isometric plantar flexion (MVIP) torque while the displacement of the distal myotendinous junction of the medial gastrocnemius and ankle joint rotation was determined using a B-mode ultrasonograph and a goniometer, respectively. The tendon force (F) was calculated from MVIP torque and the moment arm of Achilles tendon. The elongation of the Achilles tendon (delta X) was obtained from the tendon displacements and ankle joint rotation. Achilles tendon stiffness (k) was calculated by dividing F by delta X. The specific stiffness of the Achilles tendon (k(s)) was obtained from k normalized to the Achilles tendon length at rest. The cross-sectional area of the Achilles tendon (CSA) was measured at 5, 10, and 15% of the lower leg length proximal to the insertion of the Achilles tendon using a B-mode ultrasonography. The results showed that more distal portion of the Achilles tendon had a larger CSA, and that there was a strong correlation between the average and minimum Achilles tendon CSA. delta X was 9.9 +/- 2.5 mm. k and k(s) were 330 +/- 77 N/mm and 63 +/- 20 kN, respectively. No significant correlation was seen between CSA and k(s) (r = 0.15, p > 0.05). It was suggested that a stiffer Achilles tendon did not necessarily have a thicker shape, which might indicate that the major determinant of the extendibility of the Achilles tendon was not its geometric properties in young adults. PMID- 15812148 TI - Localization and characterization of specific cornification proteins in avian epidermis. AB - Little is known about proteins involved in the formation of the stratum corneum in the avian apteric epidermis. The present immunocytochemical, autoradiographic and electrophoretic study shows that antibodies against characteristic proteins of mammalian cornification (alpha-keratins, loricrin, sciellin, filaggrin, transglutaminase) recognize avian epidermal proteins. This suggests the presence of avian protein with epitopes common to related mammalian proteins. These proteins may also be involved in the formation of the cornified core and cell envelope of mature avian corneocytes. The immunoblotting study suggests that protein bands, cross-reactive for antibodies against loricrin (45, 52-57 kDa), sciellin (54, 84 kDa), filaggrin (32, 38, 45-48 kDa), and transglutaminase (40, 50, 58 kDa), are present in the avian epidermis. Immunocytochemistry shows that immunoreactivity for the above proteins is localized in the transitional and lowermost corneous layer of apteric epidermis. Their epitopes are rapidly masked/altered in cornifying cells and are no longer detectable in mature corneocytes. In scaled epidermis a thick layer made of beta-keratins of 14-18, 20 22, and 33 kDa is formed. Only in feathered epidermis (not in scale epidermis), an antifeather chicken beta-keratin antibody recognized a protein band at 8-12 kDa. This small beta-keratin is probably suitable for the formation of long, axial filaments in elongated barb, barbule and calamus cells. Conversely, the larger beta-keratins in scales are irregularly deposited forming flat plates. Tritiated histidine coupled to autoradiography show an absence of both keratohyalin and histidine-rich proteins in adult feathered and scaled epidermis. Most of the labeling appears in proteins within the range of beta- and alpha keratins. These data on apteric epidermis support the hypothesis of an evolution of the apteric and interfollicular epidermis from the expansion of hinge regions of protoavian archosaurians. PMID- 15812149 TI - Anterograde tracer study on the nucleus geniculatus dorsalis and its internal synaptic structure in chick brain. AB - In the present study the terminals of retinal fibres and those of internal layer cells in ventral geniculate nucleus of chicks were labelled with the anterograde tracer biotinylated dextran amine. The tracer showed the connections from the internal cell layers of ventral geniculate nucleus to the medial part of the dorsal lateral geniculate nucleus. The labelled retinal terminals were located exactly in the lateral part of nucleus. The labelled terminals in the two parts of the nucleus were analysed with the electron microscope and showed a different synaptic organisation in the two parts of the dorsal lateral geniculate nucleus. In the lateral part, two kinds of synaptic glomeruli were found mostly in the vicinity of large dendrites, which are proximal dendrites of projection neurons. One type is a simple glomerulus containing a large dendrite, a large optic terminal and a large and/or series of asymmetrical synapses surrounded by glial processes. The other type is a complex synaptic unit with several pre- and postsynaptic components, among them synapses of GABA-positive axon terminals and/or dendraxons. No glomeruli were found in the medial part of the nucleus. In the medial part of the lateral geniculate nucleus, the terminals of internal layer cell axons established asymmetrical synapses with dendrites. Often, a large terminals and large dendritic profiles established serial asymmetrical synapses. GABA-positive myelinated fibres entered and ramified in both parts of the dorsal lateral geniculate nucleus, and GABA-positive terminals were seen to form synapses on the same dendrite near to the asymmetrical contacts. To our knowledge, this is the first report of the connection from ventral geniculate internal layer cells to the dorsal lateral geniculate nucleus in the chick. PMID- 15812150 TI - The tenth vascular component in a rare form of the cerebral arterial circle of fetuses. AB - The ancient knowledge of the cerebral arterial circle (CAC) angioarchitecture could be supplemented by either a new or a forgotten old fact about it. Therefore, 7 cases of a rare form of the CAC of fetuses were described, where the so-called intermediate communicating artery, as an excess vessel into the posterior part, represented either its new vascular component or the developmental segment of this part. The paper aims at revealing the fetal history and morphological features of this 'underdeveloped' posterior part of the CAC during the 2nd and 3rd gestational trimesters, which could be of anatomical and clinical meaning in similar prenatal and/or postnatal cases. PMID- 15812151 TI - Gastric cancer: who is at risk? AB - Gastric cancer is a multifactorial disease. Helicobacter pylori infection, host genetic factors and dietetic factors play an important role in the development of gastric cancer. Individuals with a positive family history of gastric cancer and/or pro-inflammatory polymorphisms of the interleukin-1 and tumor necrosis factor A genes infected by H. pylori virulent strains (cagA-, vacA s1-, vacA m1- and babA2-positive) have the highest risk of gastric cancer development. Diets rich in salted and smoked food and poor in fresh fruit and vegetables favor gastric carcinogenesis. Genetic combined with bacterial and host genotyping may allow for the identification of patients at high risk of gastric cancer who can benefit from preventive eradication therapy. PMID- 15812152 TI - Pathogenesis of pre-neoplastic lesions of the stomach: targets for prevention. AB - Gastric atrophy and intestinal metaplasia are generally considered to be precancerous lesions of the stomach. Chronic Helicobacter pylori infection is one the most important factors in the development of these pre-malignant gastric lesions. In addition to bacterial factors, polymorphisms in the cytokine genes of the host that modulate inflammatory responses are found to have a synergistic effect in the development of gastric cancer as well as pre-neoplastic lesions. Recently, inappropriate activation of the intestine-specific transcription factor like the homeobox gene complex CDX1 and CDX2 are found to be an important contributing factor in the induction of intestinal metaplasia in the stomach. Aberrant expression of cyclooxygenase-2 and epigenetic changes are also frequently detected in pre-neoplastic gastric lesions. One of the most important questions relating to these pre-neoplastic gastric lesions is that whether H. pylori eradication could reverse these changes. However, most controlled studies showed no or just modest improvement in intestinal metaplasia after H. pylori eradication. Further studies should evaluate the role of other chemopreventive agents, particularly cyclooxygenase-2 inhibitor, on regression of pre-neoplastic lesions. PMID- 15812153 TI - Prevention of gastric cancer by Helicobacter pylori eradication. AB - The evidence supporting the important role of Helicobacter pylori causing gastric cancer is getting stronger. The mechanisms by which H. pylori can influence the progression to severe changes in the gastric mucosa are under investigation. An increased gastric epithelial cell proliferation has been observed in individuals infected with H. pylori. This lifelong increased cell turnover is deemed to be a major risk factor for increased mutational changes and may lead to the development of gastric cancer. Successful eradication of H. pylori infection induces the healing of the gastritis and a significant decrease in gastric epithelial cell proliferation. Nevertheless, it is right now unknown at which time the point of no return, meaning at which time an eradication therapy leads to a benefit for the individual to prevent gastric cancer, has been reached. Therefore the major question that arises is to whom an eradication therapy should be offered to prevent gastric cancer. A general elimination of the infection might be worthwhile, but seems to be unrealistic now because of the high prevalence of the infection and the missing of a vaccine. This review reflects possible mechanisms of gastric cancer development induced by chronic H. pylori infection and recent investigational trials for prevention of gastric cancer by H. pylori eradication therapy will be discussed. PMID- 15812154 TI - Chemoprevention of gastric cancer: role of COX-2 inhibitors and other agents. AB - Despite the decrease in incidence, gastric cancer remains the second leading cause of cancer-related death worldwide. Prevention is likely to be the most effective means of not only reducing the incidence but also mortality from this disease. The term 'chemoprevention' has been referred to the prevention of cancer using specific agents to suppress or reverse the carcinogenic process. In recent years, attention has been focused on the anticancer properties of non-steroidal anti-inflammatory drugs (NSAIDs), Helicobacter pylori eradication therapy and diet life-style. In vitro and in vivo studies show that widespread and long-term use of NSAIDs may be adopted in the healthy population for gastric chemoprevention. Albeit, enthusiasm has been thwarted by the potential toxic effects, i.e., risk of peptic ulcer disease. The new NSAIDs, selective cyclooxygenase-2 inhibitors, causing less injury to the mucosa of the upper gastrointestinal tract may be a valid alternative. However, fundamental questions such as safety, efficacy, mechanisms of actions, and optimal treatment regimens need to be defined. H. pylori triggers gastric carcinogenesis, however, cost effect analyses suggest that only a subgroup of H. pylori-infected subjects present beneficial changes following eradication therapy. Diet plays an important role in the pathogenesis of gastric cancer either increasing the risks of or protecting against, cancer development. Thus, a reasonable suggestion for the general population is a natural chemoprevention based on life-style 'eat to live, not live to eat'. PMID- 15812155 TI - New diagnostic approaches for early detection of gastric cancer. AB - Detection of gastric cancer in early stages is vitally important for ascertaining better prognosis and quality of life for the patients. Therefore, every endoscopist should be trained to master enough diagnostic skills to identify early gastric cancer that often shows minimal alteration from the surrounding mucosa, easily evading detection. For the first step, it is essential that endoscopists understand the normal gastric mucosa as well as the mucosal changes caused by chronic H. pylori infection, a high-risk condition for the development of gastric cancer. Once a suspicious lesion is identified, use of a dye-spraying method may be useful to clarify structural alteration caused by neoplastic changes and facilitate the biopsy sampling. Development of zoom (magnifying) endoscopy enabling 80x magnification with a one-touch switch from conventional endoscopic observation helps to identify the detailed surface structure as well as the vascular architecture of the mucosa without tissue biopsy. Combined with chromoendoscopy, this powerful endoscopic method can be used to identify small cancer foci or delineate the margin of early gastric cancer that can be treatable by mucosal dissection. Other new modalities using a variety of optical devices have been developed but the real value of their utility still remains to be proven in the actual clinical settings. PMID- 15812156 TI - Novel endoscopic imaging techniques toward in vivo observation of living cancer cells in the gastrointestinal tract. AB - Recent advances in endoscopic imaging technology have enabled the visualization of cellular-level microstructures of early-stage cancer and its precursors. Chromoendoscopy, magnifying endoscopy, endoscopic optical coherent tomography (EOCT) spectroscopy, and various combinations of these technologies, are all important for the recognition of small and unclear lesions. In order to observe cancer cells in vivo two types of ultra-high magnifying endoscope--'laser scanning confocal laser-scanning endoscopy series' and 'contact endoscopy series' -that have a maximum of >1,000 times magnifying power have been developed. The use of these endoscopes has allowed the generation of high quality images of both living cancer cells and normal cells in the gastrointestinal tract. In particular, clear images of cells and their nuclei, equivalent to the high quality that is possible with conventional cytology, have been produced. These novel imaging technologies may make in vivo histological diagnosis by virtual histology possible. PMID- 15812157 TI - Update on surgery of gastric cancer: new procedures versus standard technique. AB - D2 lymphadenectomy has been the mainstay of treatment for every stage of gastric cancer including early gastric cancer in Japan. However, the use of conventional D2 nodal dissection is being challenged. There was a recent improvement in techniques for preoperative diagnosis and perioperative diagnosis. Less extensive surgeries to maintain patients' quality of life have been introduced as standard treatment for some forms of early gastric cancer in the Gastric Cancer Treatment Guidelines 2001 (The Japanese Gastric Cancer Association). Superextended dissection (more than D2) for non-early gastric cancer is set at investigational treatment. Japanese surgeons are now aiming at wide variations of surgical treatment according to the stage of disease based on new procedures. Further evaluations are proceeding to prove superior to standard techniques. PMID- 15812158 TI - (Neo)adjuvant strategies of advanced gastric carcinoma: time for a change? AB - Despite surgical R0 resections, patients with gastric cancer stage UICC II-III have a high risk of recurrence and metachronic metastases. Preliminary evidence exists that adjuvant chemotherapy or neoadjuvant chemo(radio)therapy protocols may improve the prognosis of these patients undergoing surgery of gastric cancer with curative intention. As for palliative regimens, 5-fluorouracil and cisplatin are integral components of such (neo)adjuvant strategies. Upcoming cytostatic agents, i.e. irinotecan, docetaxel, oxaliplatin, and oral fluoropyridines are currently under investigation in new multimodality treatment regimens and may further increase R0 resection rates and may prolong disease-free and overall survival in the treatment of advanced localized gastric cancer. PMID- 15812159 TI - Endoscopic palliation and nutritional support in advanced gastric cancer. AB - Most of the patients with advanced gastric cancer have incurable disease at presentation and require palliative treatment to reduce symptoms as vomiting, nausea and inability to eat. Treatment options are palliative surgery and endoscopic techniques. Insertion of self-expanding metal stents is nowadays a well-established method of treating biliary and esophageal strictures and is also effective in gastric tumors. The indication and application technique are described in this review. In addition, enteral nutrition is indicated if the gastrointestinal tract functions but swallowing or mastication is compromised by disease or if it is needed to pass an obstructed area, especially in gastric tumor patients. This article reviews the enteral nutrition techniques and their clinical value for patients with advanced gastric cancer. PMID- 15812160 TI - Recent developments in chemotherapy of advanced gastric cancer. AB - Approximately 1 million individuals develop gastric cancer every year and the mortality of gastric cancer is only second to lung cancer. The poor prognosis is caused by late diagnosis of most cancers in advanced stages and the limited therapeutic options in these stages. Apart from the elucidation of underlying molecular and genetic changes in the development and progression of gastric cancers, the development of new treatment strategies is critical for the improvement of the treatment and prognosis of these patients. In this review we have summarized and critically assessed recent studies dealing with the chemotherapy of advanced gastric cancer. While the efficacy of most treatment regimens is only limited, new developments may indicate that treatment with chemotherapy may confer some benefit in the future. PMID- 15812161 TI - Treatment of peritoneal carcinomatosis in gastric cancers. AB - Peritoneal carcinomatosis may occur after curative surgery of any gastrointestinal carcinoma, but it is however the most frequent form of evolution after curative resection of gastric carcinoma and is present at the time of surgery in many cases. This locoregional extension of cancer has a poor prognosis, with a great mortality and a poor quality of life. It is sometimes considered of such a poor prognosis that patients do not go through any resection or palliative procedure. Techniques of radiotherapy, chemotherapy and intraperitoneal chemotherapy have been used with moderate clinical efficacy. Since the 1990s, intraoperative hyperthermic peritoneal chemotherapy combined with comprehensive cytoreductive surgery has been proposed to improve prognosis of patients with peritoneal carcinomatosis from gastric origin as well as carcinomatosis from colorectal origin or pseudomyxoma peritonei. PMID- 15812162 TI - Gene therapy and virotherapy of gastric cancer: preclinical results and clinical developments. AB - Despite advances in current treatment modalities, the clinical outcome of gastric cancer remains dismal. New treatment modalities are urgently required to improve the prognosis of patients with gastric cancer. Cancer gene therapy and virotherapy comprise a potential category of new therapeutics and will be discussed in this review. To date, various gene therapy strategies have been developed, but first clinical trials reported only limited therapeutic efficacy as a result of limited gene transfer efficiency. Consequently, targeted viral vectors for enhanced delivery of transgenes to tumor cells and replicative viral systems designed to replicate selectively in malignant tissue were developed. Replication-selective oncolytic viral vectors have the advantage over non replicative systems to cause pronounced bystander effect via self-perpetuating infection of adjacent cells after cytolysis of primary targeted cells. So far, clinical studies on virotherapy showed encouraging results; especially promising are combinations of virotherapy with current modes of treatment like chemo- and radiotherapy, or insertion of therapeutic genes in the viral genome such as combination with enzyme-prodrug therapy. Further research aiming to enhance anti tumor efficacy and to improve selectivity of infection and replication, will eventually lead to full realization of the therapeutic potential of (replicating) viral vector systems for gastric cancer. PMID- 15812163 TI - Recent advances in molecular diagnosis and therapy of gastric cancer. AB - Gastric cancer is the fourth most common malignancy and the second most frequent cause of cancer-related death in the world. It is often diagnosed in advanced stages when treatment options are limited, leading to a poor prognosis. During the past 15 years, much has been learnt about the molecular mechanisms of gastric carcinogenesis and the development of metastases. This coincided with the development of new techniques for functional genomics, including both transcriptomics and proteomics, which significantly improve the ability to explore new molecular alterations involved in carcinogenesis and tumor progression. An improved understanding of the molecular pathology and pathogenesis of gastric cancer may lead to a more rapid development of molecular diagnostic and patient tailored therapeutic targets. PMID- 15812164 TI - Helicobacter pylori seropositivity and atherosclerosis risk factors. AB - Certain viral and bacterial infections may contribute to the initiation and progression of atherosclerosis. The aim of this study is to determine whether Helicobacter pylori (HP) seropositivity contributes to conventional atherosclerosis risk factors in the development of an early sign of atherosclerosis: intima-media thickness (IMT) of the carotid artery. Eighty-four patients who had at least two conventional atherosclerosis risk factors and a control group of 50 patients having no risk factors for atherosclerosis were enrolled in the study. None of the patients had ever received HP eradication treatment. HP IgG antibodies were determined by enzyme-linked immunosorbent assay. Carotid artery IMT was measured 1 cm before the carotid bifurcation. Seventy-five percent of the study group was HP seropositive. HP seropositive (n=64) and seronegative (n=21) groups were identical in terms of sex distribution, smoking pattern, mean age, hemoglobin, leukocyte, platelet, C reactive protein, erythrocyte sedimentation rate, glucose, cholesterol, triglyceride, low-density lipoprotein, high-density lipoprotein, systolic blood pressure and diastolic blood pressure levels. There was no significant difference between the mean carotid IMT of HP seropositive (0.8+/-0.3 mm) and negative (0.8+/-0.3 mm) patients in the study group. Similar to the study group, there was no statistically significant difference between mean carotid IMT of HP seropositive (0.56+/-0.19 mm) and negative patients (0.67+/-0.13 mm) in the control group (p=0.2). Future studies concerning virulent strains are needed to determine the probable role of HP in atherosclerosis. PMID- 15812165 TI - Down-regulation of secretory leukocyte protease inhibitor expression in gastric mucosa is a general phenomenon in Helicobacter pylori-related gastroduodenal diseases. AB - BACKGROUND: Secretory leukocyte protease inhibitor (SLPI) represents a multifunctional protein of the gastric mucosa exerting anti-microbial and anti inflammatory effects. Recently, a local down-regulation of antral SLPI expression in Helicobacter pylori (Hp)-infected healthy volunteers was demonstrated. AIM: To analyze mucosal SLPI expression in patients with various gastroduodenal disorders. METHODS: The prospective study included 90 patients with following gastroduodenal disorders diagnosed: gastric cancer (GC, n=22), duodenal ulcer (DU, n=17), Hp-positive dyspeptic patients (NUD, n=31) and Hp-negative NUD (n=20). During esophagogastroduodenoscopy, biopsies were taken each from antrum, corpus and tumor. SLPI expression was analyzed by quantitative RT-PCR and ELISA. RESULTS: Antral SLPI levels were reduced in all Hp-infected patients (NUD, DU, GC) by about 75% (1,494-1,826 pg/50 microg protein) compared to Hp-negative NUD (6,563 pg/50 microg protein, p<0.001, ANOVA). Tumor tissue had twofold higher SLPI levels than surrounding tumor-free gastric mucosa (3,900 vs. 1,826 pg/50 microg protein, p=0.013), but revealed reduced SLPI levels compared to Hp negative NUD patients (p=0.067). No differences were found between SLPI expression of intestinal and diffuse GC. SLPI transcript levels were unchanged throughout all groups and locations implying that transcriptional regulation of SLPI is not involved. CONCLUSION: Local down-regulation of SLPI in antral mucosa is a general phenomenon of Hp-related diseases. PMID- 15812166 TI - Subtotal esophageal resection in motility disorders of the esophagus. AB - BACKGROUND: Esophagectomy for motility disorders is performed infrequently. It is indicated after failed medical therapy, pneumatic dilation, non-resecting surgical and redo procedures. Patient selection in this group is challenging and the operative risk has to be weighted carefully against the poor quality of life with persistent or recurrent dysphagia. PATIENTS AND METHODS: Between September 1985 and April 2004, subtotal esophageal resections for advanced esophageal motility disorders of the esophagus not responding to previous therapy were carried out in 8 patients (6 females, 2 males). The median age of these patients was 59.5 (43-78) years. Six patients had a megaesophagus secondary to achalasia; 1 patient had a non-specific esophageal motility disorder with a stenosis of the distal esophagus, and a further patient displayed a recurrent huge epiphrenic diverticulum, which occurred in the context of a collagen disease. A transhiatal esophageal resection was performed in 6, a transthoracic procedure in 2 patients. RESULTS: Outcome assessment was done after a follow-up of 43.5 (3-92) months in median. The resection and reconstruction of the esophagus in advanced and decompensated esophageal motility disorders led to a marked functional improvement with disappearance of dysphagia. Despite previous therapeutic failures, alimentation could be restored in all patients. CONCLUSION: Favourable long-term results with significant improvement of symptoms can be achieved by esophageal resection even if endoscopic therapy or non-resecting surgical measures are unsuccessful. Transhiatal esophagectomy with gastric pull-up should be the preferred procedure and can be performed with low morbidity. PMID- 15812167 TI - Lack of serum oligoclonal antibody responses to Chlamydophila pneumoniae in multiple sclerosis. AB - Chlamydophila pneumoniae has been proposed as an aetiological agent in MS via a mechanism involving molecular mimicry. We undertook to investigate whether the presence of CSF oligoclonal IgG OCB or oligoclonal bands correlated with serum IgG raised against C. pneumoniae. Paired serum and CSF of 19 MS patients and 27 control patients with other neurological diseases were studied by IEF and Western blotting. Only 1 of 19 MS patients had serum antibodies against C. pneumoniae compared with 2 of the 26 control patients. This was not significant, leading us to conclude that this study does not support the theory of an association between C. pneumoniae and MS. PMID- 15812168 TI - Computer-assisted phenotype characterization for genetic research in psychiatry. AB - Psychiatric disorders differ from other complex phenotypes in their lack of objectively assessable biological markers that contribute to the establishment of a research diagnosis for genetic studies. To nevertheless allow for the delineation of genetically meaningful diagnostic entities for psychiatric genetic research, comprehensive phenotype characterization procedures are required. It is widely agreed that these should include the standardized assessment of life-time clinical symptomatology, sociodemographic, and environmental factors. Data should be based on several sources, i.e. diagnostic interviews with probands and their relatives as well as a thorough review of medical records, and final assignment of diagnosis should follow robust algorithms (i.e. best-estimate procedures, consensus diagnosis). Here, we outline a practical implementation of such a phenotype characterization strategy, including patient recruitment, study enrolment procedures, comprehensive diagnostic assessment, and data management. We argue that successful psychiatric phenotype characterization requires flexible tools. For this purpose, we have developed a computer-assisted phenotype characterization inventory, built around the backbone of a relational database. It allows for the straightforward assessment of symptoms, automated error checks and diagnostic assignment, easily manageable data storage and handling, and flexible data transfer between various research centers even across language barriers, while at the same time keeping up with the highest standards for the protection of sensitive patient data. PMID- 15812169 TI - Defining the phenotype in human genetic studies: forward genetics and reverse phenotyping. AB - The definition of phenotypes for genetic study is a challenging endeavor. Just as we apply strict quality standards to genotype data, we should expect that phenotypes meet consistently high standards of reproducibility and validity. The methods for achieving accurate phenotype assignment in the research setting--the 'research diagnosis'--are different from the methods used in clinical diagnosis in the patient care setting. We evaluate some of the main challenges of phenotype definition in human genetics, and begin to outline a set of standards to which phenotypes used in genetics studies may aspire with the goal of increasing the quality and reproducibility of linkage and association studies. Revisiting the traditional phenotype definitions through a focus on familial components and heritable endophenotypes is a time-honored approach. Reverse phenotyping, where phenotypes are refined based on genetic marker data, may be a promising new approach. The stakes are high, since the success of gene mapping in genetically complex disorders hinges on the ability to delineate the target phenotype with accuracy and precision. PMID- 15812170 TI - Quantifying the percent increase in minimum sample size for SNP genotyping errors in genetic model-based association studies. AB - Kang et al. [Genet Epidemiol 2004;26:132-141] addressed the question of which genotype misclassification errors are most costly, in terms of minimum percentage increase in sample size necessary (%MSSN) to maintain constant asymptotic power and significance level, when performing case/control studies of genetic association in a genetic model-free setting. They answered the question for single nucleotide polymorphisms (SNPs) using the 2 x 3 chi2 test of independence. We address the same question here for a genetic model-based framework. The genetic model parameters considered are: disease model (dominant, recessive), genotypic relative risk, SNP (marker) and disease allele frequency, and linkage disequilibrium. %MSSN coefficients of each of the six possible error rates are determined by expanding the non-centrality parameter of the asymptotic distribution of the 2 x 3 chi2 test under a specified alternative hypothesis to approximate %MSSN using a linear Taylor series in the error rates. In this work we assume errors misclassifying one homozygote as another homozygote are 0, since these errors are thought to rarely occur in practice. Our findings are that there are settings of the genetic model parameters that lead to large total %MSSN for both dominant and recessive models. As SNP minor allele approaches 0, total %MSSN increases without bound, independent of other genetic model parameters. In general, %MSSN is a complex function of the genetic model parameters. Use of SNPs with small minor allele frequency requires careful attention to frequency of genotyping errors to insure that power specifications are met. Software to perform these calculations for study design is available, and an example of its use to study a disease is given. PMID- 15812171 TI - Case-control association studies in mixed populations: correcting using genomic control. AB - OBJECTIVE: Case-control association studies in mixed populations can result in spurious disease-marker associations if subpopulation disease prevalence and marker frequencies both differ. Genomic control (GC) uses neutral loci to correct for spurious association (due to population stratification), but how well this works remains undetermined. METHODS: We simulated and mixed populations with different disease and marker frequencies but without marker-disease association. We generated case-control datasets, calculated the chi2 for disease association with each marker, and applied two GC procedures, dividing by the mean chi2 or median-chi2/0.456. RESULTS: Corrections became conservative (false positive rate [FPR] <5%) with increasing subpopulation prevalence and marker differences. The mean correction resulted in FPRs close to 5% at average subpopulation allele frequency differences <0.26, but inclusion of just a few markers with large frequency differences resulted in conservative FPRs. FPRs from the median correction were mostly conservative but became anticonservative when a few markers with large frequency differences were included. CONCLUSION: GC can both lead to a notable loss of power to detect a true association (conservative) in many circumstances or may fail to eliminate the spurious associations (anticonservative). The mean correction factor is useful in certain situations to correct population stratification, but it is difficult to know when those situations exist. PMID- 15812172 TI - Incorporating individual error rate into association test of unmatched case control design. AB - OBJECTIVES: Genotyping error commonly occurs and could reduce the power and bias statistical inference in genetics studies. In addition to genotypes, some automated biotechnologies also provide quality measurement of each individual genotype. We studied the relationship between the quality measurement and genotyping error rate. Furthermore, we propose two association tests incorporating the genotyping quality information with the goal to improve statistical power and inference. METHODS: 50 pairs of DNA sample duplicates were typed for 232 SNPs by BeadArray technology. We used scatter plot, smoothing function and generalized additive models to investigate the relationship between genotype quality score (q) and inconsistency rate (i) among duplicates. We constructed two association tests: (1) weighted contingency table test (WCT) and (2) likelihood ratio test (LRT) to incorporate individual genotype error rate (epsilon(i)), in unmatched case-control setting. RESULTS: In the 50 duplicates, we found q and i were in strong negative association, suggesting the genotypes with low quality score were more likely to be mistyped. The WCT improved the statistical power and partially corrects the bias in point estimation. The LRT offered moderate power gain, but was able to correct the bias in odds ratio estimation. The two new methods also performed favorably in some scenarios when epsilon(i) was mis-specified. CONCLUSIONS: With increasing number of genetic studies and application of automated genotyping technology, there is a growing need to adequately account for individual genotype error rate in statistical analysis. Our study represents an initial step to address this need and points out a promising direction for further research. PMID- 15812173 TI - Comparative informativeness for linkage of multiple SNPs and single microsatellites. AB - Single nucleotide polymorphisms (SNPs), or biallelic markers, are popular in genetic linkage studies due to their abundance in the genome, stability, and ease of scoring. We determined the 'information ratio' (IR) of closely spaced SNPs in simulated nuclear families and affected sib pairs (ASPs). (The IR is the ratio of actual average maximum lod score to the maximum lod score attainable if the marker were fully informative.) The nuclear families included parental information, whereas the ASPs did not. We analyzed these SNPs in two ways: (1) using multipoint analysis, and (2) treating the SNPs as 'composite markers' (i.e., haplotypes, as assigned by GENEHUNTER). (3) We also calculated the IR of a single microsatellite marker with multiple alleles and compared with the IR from the SNPs. For each set of input conditions, we simulated 1000 nuclear families, of 2, 3, 4, or 5 children each, as well as 1000 ASPs. We generated SNP marker data for strings of k = 1, 2, 3, 5, 7, and 10 SNP loci, with no recombination (theta = 0) and no linkage disequilibrium among the SNPs. The MAF (minor allele frequency) was either 0.5 or 0.25, and allele frequencies were the same for all k loci in any analysis. We also generated marker data for one single-locus microsatellite marker, with m = 3, 4, 5, 6, 7, and 9 equally frequent alleles. In all simulations, the disease was fully penetrant dominant, and there was no recombination or linkage disequilibrium among markers or between marker and disease. When multipoint analysis was used, we found that 5-7 closely spaced SNPs were usually enough to yield an IR of approximately 100%, for nuclear families of any size. However, for the ASPs, even 7-10 SNPs yielded an IR of only 70-80%. A microsatellite with 9 equally frequent alleles yielded about the same IR (86-88%) as a string of 4-5 SNPs, in nuclear families. SNPs analyzed as 'composite markers' analyses performed worse, due to the inherent ambiguity of SNP haplotyping. PMID- 15812174 TI - Issues in association analysis: error control in case-control association studies for disease gene discovery. AB - Several sources of errors are discussed. While genotyping errors have little effect on power in case-control association studies, they tend to strongly increase false positive results in TDT type tests unless occurrence of errors is allowed for in the analysis (e.g., TDTae test). Disregarding non-genetic risk factors is shown to lead to a form of hidden heterogeneity, which can strongly reduce power. Stratification of data into more homogeneous subgroups is advocated as a simple solution to allowing for non-genetic risk factors such as socio economic status and food preferences. PMID- 15812175 TI - Comparative study of multipoint methods for genotype error detection. AB - Several programs are currently available for the detection of genotyping error that may or may not be Mendelianly inconsistent. However, no systematic study exists that evaluates their performance under varying pedigree structures and sizes, marker spacing, and allele frequencies. Our simulation study compares four multipoint methods: Merlin, Mendel4, SimWalk2, and Sibmed. We look at empirical thresholds, power, and false-positive rates on 7 small pedigree structures that included sibships with and without genotyped parents, and a three-generation pedigree, using 11 microsatellite markers with 3 different map spacings. Simulated data includes 5,000 replicates of each pedigree structure and marker map, with random genotyping errors in about 4% of the middle marker's genotypes. We found that the default thresholds used by these programs provide low power (47 72%). Power is improved more by adding genotyped siblings than by using more closely spaced markers. Some mistyping methods are sensitive to the frequencies of the observed alleles. Siblings of mistyped individuals have elevated false positive rates, as do markers close to the mistyped marker. We conclude that thresholds should be decided based on the pedigree and marker data and that greater focus should be placed on modeling genotyping error when computing likelihoods, rather than on detecting and eliminating genotyping errors. PMID- 15812176 TI - Optimal selection of SNP markers for disease association studies. AB - Genetic association studies with population samples hold the promise of uncovering the susceptibility genes underlying the heritability of complex or common disease. Most association studies rely on the use of surrogate markers, single-nucleotide polymorphism (SNP) being the most suitable due to their abundance and ease of scoring. SNP marker selection is aimed to increase the chances that at least one typed SNP would be in linkage disequilibrium (LD) with the disease causative variant, while at the same time controlling the cost of the study in terms of the number of markers genotyped and samples. Empirical studies reporting block-like segments in the genome with high LD and low haplotype diversity have motivated a marker selection strategy whereby subsets of SNPs that 'tag' the common haplotypes of a region are picked for genotyping, avoiding typing redundant SNPs. Based on these initial observations, a plethora of 'tagging' algorithms for selecting minimum informative subsets of SNPs has recently appeared in the literature. These differ mostly in two major aspects: the quality or correlation measure used to define tagging and the algorithm used for the minimization of the final number of tagging SNPs. In this review we describe the available tagging algorithms utilizing a 3-step unifying framework, point out their methodological and conceptual differences, and make an assessment of their assumptions, performance, and scalability. PMID- 15812177 TI - Transactivation properties of wild-type and mutant androgen receptors in transiently transfected primary human fibroblasts. AB - BACKGROUND: Stromal cells play key roles during androgen-mediated male sexual differentiation. Our objective was to establish a transient transfection method for primary human fibroblasts enabling functional characterization of wild-type (wt) and mutant androgen receptor (AR) plasmid constructs, corresponding to partial and complete androgen insensitivity syndrome (PAIS/CAIS). METHODS: An AR negative fibroblast strain (ARD842) was established from the gonads of a CAIS patient. Wt-AR or either mutants L712F (PAIS), R774C or V866M (CAIS) were transfected using a polyamine-based procedure. Alternatively, two AR-positive male foreskin fibroblast strains were investigated. Androgen-induced activation of two co-transfected reporter plasmids ((ARE)(2)TATA-, MMTV-luciferase) was measured. RESULTS: All three fibroblast strains showed a ligand-dependent rise of luciferase activity after transfection of wt-AR. Mutant plasmids were assessed in AR-negative ARD842 cells. While L712F showed high partial activity, R774C and V866M were nearly inactive. The intrinsic AR of normal foreskin fibroblasts revealed no measurable ligand-inducible reporter gene activity. CONCLUSIONS: Polyamine-based transfection of AR plasmids into cultured fibroblasts provides a promising tool for analysis of AR transactivation, thereby considering a stromal cellular background. This is supported by the mutant ARs which showed the expected levels of impaired transactivation with respect to the corresponding AIS phenotypes. The role of the intrinsic AR in normal male human foreskin fibroblasts needs further exploration. PMID- 15812179 TI - Removal of hepatitis C virus by G-1 beads in sera from patients with chronic hepatitis C. AB - Recently, a new method of extracorporeal granulocyte depletion apheresis has been developed to treat inflammatory systemic diseases using an Adacolumn (Japan Immunoresearch Laboratories, Takasaki, Japan) that is filled with acetate cellulose beads (G-1 beads) to adsorb the granulocytes. We examined whether hepatitis C virus (HCV) is adsorbed after incubation of the Adacolumn with the sera from patients with HCV-RNA-positive chronic hepatitis C. PATIENTS AND METHODS: A total of 10 patients with chronic hepatitis C, whose levels of HCV RNA were greater than 800 kIU/ml were examined. The serum was incubated with 500 G-1 beads in a syringe at 37 degrees C for 1 h. After removal of the serum, the beads were washed with RNase-free water. The G-1 beads were removed from the syringe after centrifugation. RNA was extracted from 200 microl of the wash waste and from 10, 50, 100 and 200 beads, respectively, using TRIZol regent. Detection of HCV RNA was performed using the nested PCR method. RESULTS: HCV RNA was detected from as few as 10 G-1 beads. HCV RNA was not detected from waste fluid collected after the last wash from any of the patients. Further, HCV RNA was detected in the initial waste fluid after the 37 degrees C incubation with serum in all of the patients. Since HCV RNA was detected on the G-1 beads, but not from the last washing solution in the current examination, these results suggest that the G-1 beads adsorbed HCV RNA. CONCLUSIONS: Our in vitro study confirmed that G-1 beads adsorbed HCV; therefore, apheresis using a column filled with G-1 beads may reduce the HCV RNA load in the blood of patients with chronic hepatitis C. PMID- 15812178 TI - Alteration in gene expression profile by full-length hepatitis B virus genome. AB - Persistent expression of hepatitis B virus (HBV) proteins is thought to be involved in virus-related hepatocarcinogenesis. Here, we compared the gene expression profile of cells persistently expressing the full-length HBV with that of negative control cells to comprehensively investigate virus-mediated changes in the gene expression of the host cells. RNA samples from both virus-expressing and negative control cells were used for the DNA array assay. DNA array assay and subsequent corroboration assays revealed that expression of 14 of 1,176 genes (1.2%) was altered in response to virus expression. The upregulated genes included CD44, high mobility group protein-I, thymosin beta-10 and 27-kD heat shock protein, while the downregulated genes included NM23-H1, all of which are thought to be associated with the development or progression of carcinoma in the liver or other organs. Furthermore, virus expression resulted in the decrease of two apoptosis-inducing molecules, caspase-3 and BAX, which may also contribute to carcinogenesis through prolonged survival of the host cell. Thus, expression of the virus genome caused carcinogenesis-related changes in host cell gene expression. HBV expression may change the host cell to a malignant phenotype through alterations in the expression levels of a set of genes. PMID- 15812180 TI - Molecular characterization of Jurkat cells persistently infected with vaccinia virus mutant vp811. AB - Previous reports from this laboratory demonstrated that persistent infections could be established in Jurkat cells with wild-type vaccinia virus (Jvac). The infected cells elicit increased expression of IL-2, IL-2Ralpha and IL-6 but not of IL-1beta nor of interferon-gamma. The persistently infected cells are also capable to transactivate the LTR gene of HIV-1 virus (HIV-1LTR). In addition, NFkappabeta and NFAT are found activated in Jvac. To find out whether the gene(s) responsible for establishment, maintenance and molecular characteristics of persistently infected cells are located in the terminal ends of the molecule, Jurkat cells were infected with a deletion mutant Vp811 that lacks 32.7 and 14.9 kb at the left and right terminal, respectively. The results indicate that the deletion mutant is capable of establishing persistent infections. The persistently infected cells show a similar pattern of expression to that observed in Jvac cells. It is concluded that the viral gene(s) involved in persistence and molecular cellular changes is (are) present in the mutant. PMID- 15812181 TI - Characterization of nonpolio enteroviruses recovered from patients with aseptic meningitis in Korea. AB - OBJECTIVES: We attempted to characterize nonpolio enteroviruses recovered from Korean patients with aseptic meningitis. METHODS: We performed RT-PCR on the 5' nontranslated region using clinical specimens. Infectious clinical isolates were amplified by infecting Vero cells with RT-PCR-positive clinical specimens. We then investigated the direct effect in primary neuronal cells or cardiomyocytes following virus infection. RESULTS: Total 12 clinical isolates were subtypically analyzed by both RT-PCR/sequencing comparison of the VP-1 region and neutralization assay. 43-2, 43-2S, 57 and 58 were found to be coxsackievirus B1 (CVB1), 312 to be CVB5, 14-2S and 327 to be echovirus 6, 165 to be echovirus 9, 337 to be echovirus 11, and 270 to be echovirus 30. All the clinical isolates tested showed profound cytotoxicity to various degrees in the primary neuronal cells within 24 h postinfection at 10 MOI. By contrast, a significant cytopathic effect was observed in the primary cardiomyocytes at 3-5 days postinfection at 50 MOI. CONCLUSIONS: The present study suggests that the clinical isolates recovered from Korean patients belonged to different CVB or echovirus serotypes and that these viruses showed diversities in their virulence in primary neuronal cells and cardiomyocytes. PMID- 15812182 TI - Analysis of the 5' end structure of HCV subgenomic RNA replicated in a Huh7 cell line. AB - OBJECTIVE: Recently, HCV subgenomic RNA that replicates in vitro in a certain cell line have been elucidated. Since the 5' end of the genome of positive strand RNA viruses is often modified with a cap structure or a covalently linked protein, we have assessed structural feature of the HCV genome obtained from Huh7 cells in which HCV subgenomic RNA has been shown to efficiently self-replicate. METHODS: HCV subgenomic RNA was obtained from the Huh7 and was analyzed for its 5' end. RESULTS: Phosphorylation of the genomic RNA by polynucleotide kinase was observed only after treatment with phosphatase. The labeling efficiency of the genome with polynucleotide kinase was not enhanced by treatment with pyrophosphatase. CONCLUSION: It is suggested that the 5' end of HCV genomic RNA obtained from HCV replicon cells is not modified except phosphorylation. Furthermore, analysis of the 5' end of the HCV RNA obtained from the HCV subgenome self-replicating cells revealed the presence of two types of subgenomic RNA that contained either guanylate or adenylate at the 5' end. This result indicates that the 5' end of the subgenome in Huh7 cells is redundant and there is no significant evolutionary advantage between the two genomes. PMID- 15812183 TI - Infectious source factors affecting the severity of sexually transmitted acute hepatitis due to hepatitis B virus genotype C. AB - OBJECTIVE: The aim of this study was to identify clinical features and virological aspects of infectious sources that are related to the severity of sexually transmitted acute hepatitis B virus (HBV) infection in patients, especially in cases of genotype C. METHODS: Nineteen patients with acute HBV infection, 10 classified with severe acute hepatitis (SH) (prothrombin time; PT <40%) and 9 with typical acute hepatitis (AH) (PT >40%), and their infectious sources (all were sexual partners) were studied. Infectious source factors were analyzed in relation to the severity of hepatitis in the patients' partners. RESULTS: The nucleotide homology of HBV-DNA between each pair was >/=98.9%. Sixteen were infected with HBV genotype C. Among the 16 infectious sources, age, numbers with elevated alanine aminotransferase (ALT, 7/9 vs. 1/7), anti-HBe positivity (8/9 vs. 1/7) and core promoter mutations at nt 1762 (7/9 vs. 1/7), nt 1764 (8/9 vs. 1/7) and precore mutation at nt 1896 (8/9 vs. 1/7) were significantly higher in the sources of SH than in those of AH. CONCLUSION: Higher age, elevated ALT, anti-HBe positivity and core promoter/precore mutations were possible risk factors for an infectious source of the severe form of sexually transmitted acute hepatitis due to HBV genotype C. PMID- 15812184 TI - Early dynamics of hepatitis C virus in the circulation of chimpanzees with experimental infection. AB - Two chimpanzees were inoculated with hepatitis C virus (HCV) and followed on a daily basis for 12 days. HCV RNA became detectable in their sera on day 5 by polymerase chain reaction with the detection limit of 10(2) copies/ml. Based on an exponential growth observed until 8 or 9 days after inoculation in their sera, the doubling time of HCV in the circulation was estimated at 6.3-8.6 h and log time (time required to grow 10-fold) at 31.3- 42.9 h. The exact doubling time of HCV determined in them would help plan an efficient strategy for screening out blood donors in the window period of infection between the exposure and the development of antibody to HCV in serum. PMID- 15812185 TI - SARS-CoV infection was from at least two origins in the Taiwan area. AB - OBJECTIVE: Severe acute respiratory syndrome (SARS) is caused by a new coronavirus. Genomic sequence analysis will provide the molecular epidemiology and help to develop vaccines. METHODS: We developed a rapid method to amplify and sequence the whole SARS-CoV genome from clinical specimens. The technique employed one-step multiplex RT-PCR to amplify the whole SARS-CoV genome, and then nested PCR was performed to amplify a 2-kb region separately. The PCR products were sequenced. RESULTS: We sequenced the genomes of SARS-CoV from 3 clinical specimens obtained in Taiwan. The sequences were similar to those reported by other groups, except that 17 single nucleotide variations and two 2-nucleotide deletions, and a 1-nucleotide deletion were found. All the variations in the clinical specimens did not alter the amino acid sequence. Of these 17 sequenced variants, two loci (positions 26203 and 27812) were segregated together as a specific genotype - T:T or C:C. Phylogenetic analysis showed two major clusters of SARS patients in Taiwan. CONCLUSION: We developed a very economical and rapid method to sequence the whole genome of SARS-CoV, which can avoid cultural influence. From our results, SARS patients in Taiwan may be infected from two different origins. PMID- 15812186 TI - Hepatitis B virus genotypes in Korea: an endemic area of hepatitis B virus infection. AB - OBJECTIVES: It has been reported that distribution of hepatitis B virus (HBV) genotypes shows geographic difference and are associated with clinical outcomes of HBV infection, including response to antiviral therapy and progression of chronic liver diseases. In this study, we analyzed the distribution of HBV genotypes according to the various clinical outcomes of chronic HBV infection in Korea, which is one of the most endemic areas of HBV infection. METHODS: A total of 200 patients with chronic HBV infection were enrolled. Clinical diagnoses of the 200 patients with chronic liver diseases were as follows: hepatitis B e antigen (HBeAg)-positive healthy carrier (defined as HBeAg(+), anti-HBe(-), HBV DNA(+) by hybridization, normal transaminase; n = 40); inactive HBsAg carrier (n = 40); chronic hepatitis B (n = 40); liver cirrhosis (n = 40); hepatocellular carcinoma (n = 40). HBV genotypes were determined by nested polymerase chain reaction using genotype-specific primers. RESULTS: All patients except 2 (inactive HBsAg carriers) were positive for nested PCR and they have genotype C regardless of clinical outcomes. CONCLUSIONS: HBV genotype was genotype C regardless of various clinical outcomes of chronic HBV infection in Korea. Considering that HBV genotypes have clinical relevance, distribution of HBV genotype in each area should be monitored when management for chronic HBV infection is planned. PMID- 15812187 TI - Significance of serum ribavirin concentration in combination therapy of interferon and ribavirin for chronic hepatitis C. AB - OBJECTIVE: The purpose of this clinical retrospective cohort study was to determine the most suitable ribavirin concentration on combination therapy of interferon (IFN)-ribavirin. METHODS: Entry criteria were serum HCV-RNA level >/=100 KIU/ml, HCV-genotype 1b, chronic hepatitis, and initial combination treatment of IFN-alpha-2b (6 million units daily for 2 weeks and then 3 times weekly for 6 weeks) and ribavirin (600-800 mg/day) for 8 weeks without stopping or decreasing the dosage of IFN and/or ribavirin. Sixty-eight consecutive patients who satisfied the above criteria were given maintenance therapy for another 16 weeks. RESULTS: A sustained virological response (SVR) rate of 25.0% (17/68) was seen in all subjects. The SVR rate was 44.0% (11/25) in the high ribavirin group with a serum ribavirin concentration of >/=3,000 ng/ml at 8 weeks after initiation of combination therapy. SVR was significantly dependent at a serum ribavirin level of >/=3,000 ng/ml (p = 0.005). The incidence of discontinuations and dose modifications for combination therapy in patients having a serum ribavirin concentration of >/=3,500 ng/ml 8 weeks after initiation of therapy was 57.1% (4/7). This value was statistically higher than that in patents with <3,500 ng/ml (p = 0.033). CONCLUSION: Our results showed that the most suitable serum ribavirin concentrations are from 3,000 to 3,500 ng/ml 8 weeks after initiation of combination therapy. PMID- 15812188 TI - Langerhans cells stimulated by mechanical stress are susceptible to measles virus infection. AB - OBJECTIVE: Measles virus (MV) first infects the human respiratory tract, but the initial target cells are unknown. We examined whether MV infects Langerhans cell like dendritic cells (LCs) generated from CD14(+) monocytes in the presence of GM CSF, IL-4, and TGF-beta1. METHODS: Cultured LCs were established as described recently [Biochem Biophys Res Commun 2003;306:674-679]. The expression of immunological markers was detected by FACScan. Infection with MV was assessed by syncytia formation, viral-specific fluorescence, and Western blotting. RESULTS: MV did not infect and replicate the freshly established, unstimulated LCs expressing CD1a, E-cadherin and Langerin but not CD83. Also, CD150, a receptor for MV was not expressed on the surface of the LCs. However, LCs stimulated by mechanical stress such as washing and centrifugation became susceptible to MV infection. CONCLUSION: A subset of mechanically stimulated LCs but not unstimulated immature ones became susceptible to MV. The actual role of Langerhans cells in local immunity seems to be to suppress unfavorable reactions initiated by virus intrusion. PMID- 15812189 TI - Distribution of viral RNA in mouse tissues during acute phase of coxsackievirus B5 infection. AB - OBJECTIVE: To investigate histopathological changes and distribution of coxsackievirus B5 (CVB5) RNA in mouse heart, liver, and pancreas during the acute phase of infection. METHODS: C3H/HeJ male mice, aged 3-4 weeks, were inoculated intraperitoneally with 5 x 10(5) plaque-forming units of CVB5 and sacrificed at 1, 2, 3, 4, 7 and 10 days postinfection (p.i.). Inflammation of the heart, liver, and pancreatic tissue sections was evaluated by hematoxylin and eosin staining, and virus was detected using antibody to viral coat protein VP1. A quantitative real-time RT-PCR method, using primers and probe targeted to the highly conserved sequences in the 5'-untranslated region of the virus, was used to evaluate the kinetics of CVB5 RNA during the development of myocarditis or pancreatitis. RESULTS: Marginal inflammatory changes were observed in the heart tissues although viral RNA was constantly present between 1 and 10 days p.i., peaking at 4 days p.i. The pancreatic tissues displayed massive lymphocyte infiltration and loss of acinar cells at day 4 p.i. and viral RNA was detected between 1 and 10 days p.i., peaking at 2-3 days p.i. In the liver, viral RNA was detected between 1 and 7 days. No mortality was observed. CONCLUSIONS: CVB5 induced acute pancreatitis without subsequent development of myocarditis. Clearance of CVB5 RNA from the pancreas and heart was slower than clearance from the liver. Our real time RT-PCR method, which is more sensitive than conventional plaque assay, may provide valuable insight into viral RNA kinetics during CVB5 infection. PMID- 15812190 TI - Suckling mice were used to detect infectious dengue-2 viruses by intracerebral injection of the full-length RNA transcript. AB - OBJECTIVE: In this study we established a mouse brain injection system to detect infectious dengue-2 virus produced from the full-length RNA transcripts. METHODS: In vitro transcription was used to synthesize full-length dengue-2 virus RNA from the plasmid pRS424FLDEN2NGC, which was intracerebrally injected into the 6-day old suckling mice (ICR strains). Engineered dengue-2 viruses were detected in the brain sections using immunohistochemistry staining. RT-PCR followed by restriction endonuclease BstEII digestion was used to confirm the mosquito C6/36 cells cocultured with the mouse brain extract. RESULTS: The mice inoculated with the full-length dengue-2 viral RNA transcript showed paralysis symptoms and died between day 10 and 13 postinjection. The dengue-2 virus-specific antigens (E, Core and NS1) were detected in all the brain and part of the liver sections of the paralyzed mice by immunohistochemistry staining, indicating the existence of dengue-2 virus in these tissues of the suckling mice. The viruses detected in the brains of suckling mice were indeed infectious, which was further confirmed by coculturing mosquito C6/36 cells with the brain extract of the injected mice. CONCLUSIONS: We developed an in vivo approach to detect and produce engineered dengue viruses with infectivity from the full-length plasmid cDNA. This suckling mice system will also aid in screening the infectious viruses that are created by site-directed mutagenesis and is useful for the studies of dengue virus gene function and pathogenesis in the host. PMID- 15812191 TI - Porcine endogenous retroviruses PERV-A and PERV-B infect neither mouse cells in vitro nor SCID mice in vivo. AB - OBJECTIVE: Porcine endogenous retroviruses (PERVs) pose a risk for xenotransplantations using pig materials as they are present in the genome of all pigs and are able to infect human cells in vitro. Until recently, transmission of PERVs in vivo was only described in severe combined immunodeficient (SCID) and nude mice inoculated with PERV-producing cells. However, in this series of experiments microchimerism could not be excluded. To overcome this problem, the risk of PERV infection was addressed in a similar way but using cell-free inoculation of mouse cells in vitro and SCID mice in vivo. METHODS: Mouse cell lines and primary cells were incubated in vitro with PERV-A, with a recombinant PERV-A/C and with PERV-B. Provirus integration was assessed by PCR. Reverse transcriptase activity was measured in the cell supernatants. SCID mice were inoculated in vivo with cell-free virus at high titers. RESULTS: None of the mouse cell lines and primary cells could be infected by PERV and no provirus integration was observed in different organs of the inoculated SCID mice. CONCLUSION: The data indicate that PERV-A, PERV-A/C and PERV-B could not infect different mouse cells. These data correlate with the recent finding that mouse cells lack a functional receptor for PERV-A. Although the receptor for PERV-B is still unknown, these data suggest that previously reported PERV transmissions to SCID and nude mice in vivo might be due to microchimerism or pseudotyping with murine viruses and indicate that normal mice are an inappropriate model for the study of PERV infection and pathogenesis. PMID- 15812192 TI - Virological and biochemical relapse after discontinuation of lamivudine monotherapy for chronic hepatitis B in Japan: comparison with breakthrough hepatitis during long-term treatment. AB - OBJECTIVE: Comparison of virological and biochemical relapse in patients with chronic hepatitis B, based on continuation or discontinuation of lamivudine monotherapy. METHODS: In Japanese genotype C-dominant hepatitis B patients, 25 patients who stopped treatment at normal levels of alanine transferase (ALT) were retrospectively compared with 75 patients who continued treatment. Both groups were matched for age, sex, and observation period after start of treatment. We investigated the relapse rates, and evaluated predictive factors for relapse and efficacy of retreatment of the discontinuous group. RESULTS: Virological and biochemical relapse occurred significantly earlier in the discontinuous than continuous group, and the peak levels and ratios of peak to pretreatment levels of serum bilirubin and ALT after relapse were not significantly different between the two groups. Multivariate analysis identified three independent factors at discontinuation of treatment associated with early biochemical relapse: HBeAg positivity, presence of liver cirrhosis, detection of basic core promoter mutant. Normalization of ALT levels with retreatment occurred in 62.5% of patients, but 2 HBeAg-positive patients retreated after the emergence of YMDD motif mutant developed severe relapse with hyperbilirubinemia. CONCLUSION: Our results in Japanese patients with genotype C-dominant hepatitis B suggest that discontinuation of lamivudine monotherapy, and retreatment after the emergence of YMDD mutant should be given attention. PMID- 15812193 TI - Replication-competent chimeric hepatitis C virus subgenomic replicons. AB - OBJECTIVE: To utilize chimeric hepatitis C virus (HCV) replicons to select adaptive mutation(s) that allow replication of a genotype 1a replicon. METHODS: We used a genetic approach to gradually apply selective pressure by generating chimeric replicons through sequential replacement of nonstructural genes of a 1b replicon with genotype 1a sequences. RESULTS: A chimeric replicon containing a genotype 1a NS5A protein did not replicate in a transient assay, but could be used to establish stable cell lines using G418 selection. The cell lines contained a K1846T mutation in NS4B which functioned as an adaptive mutation that now allowed the chimera to replicate at levels similar to wild-type replicons. Similarly, replication of a 1a NS5A5B chimera was only observed after establishment of stable cell lines, even in the presence of the K1846T mutation. Sequence analysis of this cell line revealed an additional adaptive mutation of M1496L in NS3. Lastly, by including the K1846T mutation in a replicon that was entirely genotype 1a sequence, stable 1a cell lines could be established. CONCLUSION: These studies identify an NS4B adaptive mutation, K1846T, which allows establishment of a replication-competent 1a replicon and demonstrate the utility of this chimeric approach for establishing replicons for various HCV genotypes. PMID- 15812194 TI - Hepatitis B and C virus variants in long-term immunosuppressed renal transplant patients in Latvia. AB - The incidence of genome variants of hepatitis B and hepatitis C viruses among 38 long-term (2-15 years) immunosuppressed patients after renal transplantation and 10 patients undergoing dialysis was investigated. Twelve patients had only HBV infection, 9 had only HCV infection and 14 were co-infected. Regions corresponding to the HBV X/EnII/BCP, preC/C, preS/S and to the HCV core were sequenced for molecular characterization of the HBV and HCV genomes. Fifty-seven percent of HBV DNA isolates belonged to genotype D and 42% to genotype A, whereas 77% of HCV RNA isolates belonged to genotype 1b and only 17% to genotype 3a. One sample (6%) was of genotype 2c. Detailed analysis of the above-mentioned HBV genome regions revealed the presence of nucleotide point mutations, which, in some cases, resulted in amino acid substitutions. The clinical significance of such mutations is discussed. PMID- 15812195 TI - Analysis of human cytomegalovirus UL144 variability in low-passage clinical isolates in Japan. AB - To explore a possible role for viral genes as determinants of virulence, portions of the UL144 tumor necrosis factor-like receptor gene and the UL55 envelope glycoprotein B gene from 42 patients with congenital human cytomegalovirus (HCMV) infection or other diseases were sequenced. Of the 42 patients, 16 (38%) had UL144 group 1 [group 1A, 15 of 16 (94%); group 1B, 1 of 16 (6%); group 1C, 0 of 16 (0%)], 5 patients (12%) had UL144 group 2, and 21 patients (50%) had UL144 group 3. Although group 1C was not found in Japan strains (0%), it was found in USA strains (22%). Other HCMV polymorphisms should be further evaluated for their potential relevance to neonatal infection, and acquired immunodeficiency syndrome associated HCMV diseases. PMID- 15812196 TI - Induction of metallothionein in proximal tubular cells by zinc and its potential as an endogenous antioxidant. AB - BACKGROUND: This study was undertaken to gain further insights into the expression of metallothionein (MT) in kidney, to define the necessary dosage of a metal (zinc) to achieve induction of MT and to evaluate the antioxidative potential of MT in comparison to other more common antioxidative therapeutics, like N-acetyl-L-cysteine (NAC), and endogenous molecules, like glutathione. METHODS: MT was measured in renal specimens from cadaver kidneys from patients with chronic diseases (n = 76) and controls (n = 21) by immunohistochemistry. In addition, induction experiments were performed in cell cultures of proximal tubular cells (LCC-PK1) and MT measured on the RNA and protein level (immunohistochemistry, Western and dot blotting). Antioxidative potential of MT was compared to NAC and glutathione. RESULTS: MT was restricted to tubular cells with no differences between controls and patients. Zn caused a dose-dependent increase of MT on the RNA as well as on the protein level (RNA (ratio MT/histone 3.3): control 0.34 +/- 0.12; Zn 17 microM 0.65 +/- 0.26; Zn 35 microM 1.25 +/- 0.43 (p < 0.05), Zn 52 microM 1.35 +/- 0.46 (p < 0.05), and protein: 5.8-fold increase from 47 +/- 13 mg/g total protein (n = 6) to 272 +/- 140 mg/g total protein (n = 6)). The antioxidative effect of MT was equal to NAC and glutathione. CONCLUSIONS: Induction of renal MT by zinc is easily achievable and might be an interesting therapeutic and preventive tool against oxidative stress. PMID- 15812197 TI - Introduction. PMID- 15812198 TI - Current issues in anticoagulation. AB - Anticoagulation therapy with unfractionated heparin, low-molecular-weight heparins and oral vitamin K antagonists is currently the mainstay of treatment and prevention of thromboembolic disorders (such as deep vein thrombosis, pulmonary embolism and stroke prevention in patients with atrial fibrillation). Although these therapies have proven benefits, they also have important limitations that result in their underuse in routine clinical practice. Consequently, many patients identified by guidelines as requiring anticoagulant therapy receive no or inadequate treatment. Heparins require parenteral administration and pose the risk of heparin-induced thrombocytopenia. Vitamin K antagonists have a narrow separation of antithrombotic and haemorrhagic effects and numerous food and drug-drug interactions, and require frequent coagulation monitoring and dose adjustment to ensure effective antithrombotic protection while minimizing the risk of bleeding complications. In response to these limitations, several new anticoagulants have recently been developed, including selective factor Xa inhibitors such as fondaparinux and ximelagatran, the first oral agent in the new class of direct thrombin inhibitors and the first new oral anticoagulant for almost 60 years. Ximelagatran possesses many of the properties of an ideal agent for anticoagulation therapy. With its oral formulation, consistent and predictable pharmacological profile and no coagulation monitoring, ximelagatran has the potential to increase the use and duration of anticoagulation treatment in thromboembolic disorders and to reduce the burden associated with long-term management. PMID- 15812199 TI - Ximelagatran in orthopaedic surgery. AB - Ximelagatran represents the first new oral anticoagulant since the introduction of warfarin almost 60 years ago, and has been evaluated for the treatment and prevention of a range of venous and arterial thromboembolic disorders. The MElagatran THRomboprophylaxis in Orthopaedic surgery (METHRO) and EXpanded PRophylaxis Evaluation Surgery Study (EXPRESS) studies have investigated the efficacy and safety of subcutaneous (s.c.) melagatran followed by oral ximelagatran in preventing venous thromboembolism (VTE) in patients undergoing total hip replacement or total knee replacement. In METHRO II, immediate pre operative-initiated s.c. melagatran followed by post-operative ximelagatran dose dependently reduced VTE, with the highest dose (melagatran 3 mg/ximelagatran 24 mg twice daily) associated with a significantly reduced incidence of VTE compared with the low-molecular-weight heparin (LMWH) dalteparin (15.1 vs. 28.2%; p < 0.0001). In METHRO III, the efficacy of s.c. melagatran 3 mg/ximelagatran 24 mg twice daily initiated post-operatively (4-12 h after surgery) was comparable to that of the LMWH enoxaparin initiated 12 h before surgery (total VTE incidence, 31.0 and 27.3%, respectively). Rates of severe bleeding were also comparable between treatments (melagatran/ximelagatran = 1.4%; enoxaparin = 1.7%). Treatment with melagatran/ximelagatran was significantly more effective when initiated earlier (4-8 h) rather than later (8-12 h) after surgery (total VTE incidence, 27.5 vs. 35.4%; p = 0.0034). Based on the results of METHRO II and III, the EXPRESS study evaluated the efficacy and bleeding profile of s.c. melagatran 2 mg immediately before surgery, followed by s.c. melagatran 3 mg on the evening of the day of surgery and then ximelagatran 24 mg twice daily. This regimen was significantly more effective than enoxaparin (total VTE incidence, 20.3 vs. 26.6%; p < 0.0004). Excessive bleeding (as judged by the investigator) was more frequent with melagatran/ximelagatran, but rates of fatal bleeding, critical-site bleeding and bleeding requiring re-operation did not differ between the groups. Taken together, the METHRO and EXPRESS studies demonstrate that melagatran/ximelagatran has comparable or superior efficacy to LMWHs in the prevention of VTE in orthopaedic surgery patients, and that the timing and dose of melagatran is important in optimizing the balance of efficacy and bleeding risk. PMID- 15812200 TI - The role of ximelagatran in the treatment of venous thromboembolism. AB - Clinical-based evidence demonstrates that long-term oral anticoagulant therapy with the vitamin K antagonists is highly effective for the secondary prevention of venous thromboembolism (VTE). However, owing to fear of bleeding complications and the inconvenience of coagulation monitoring, many patients do not receive the required duration of treatment. This can lead to a high incidence of recurrent VTE events and has prompted the evaluation of alternative treatment strategies and the development of new anticoagulants for VTE management. For patient groups in which it is particularly difficult to maintain the target intensity of anticoagulation, low-molecular-weight heparin (LMWH) has been found to significantly reduce the risk of recurrent VTE without increasing bleeding risk. The parenteral administration of LMWH, however, is a drawback for long-term use in the outpatient setting. Long-term warfarin use at a lower intensity (international normalized ratio [INR] 1.5-2.0) has also been assessed as a possible strategy to reduce bleeding complications and the need for monitoring, but results were disappointing when compared with conventional-intensity warfarin (INR 2.0-3.0). New therapies in development that may potentially offer a more favourable benefit-risk profile and greater consistency and predictability of response include the synthetic pentasaccharides, fondaparinux and idraparinux. These par enterally administered indirect factor Xa inhibitors have a predictable pharmacokinetic profile, allowing use without coagulation monitoring. Fondaparinux to date has only been evaluated in the initial treatment (5-7 days) of symptomatic deep vein thrombosis. In contrast, idraparinux, with its longer half-life (80 h) allowing once-weekly parenteral dosing, has the potential for long-term treatment and is currently being assessed in phase III trials for the secondary prevention of VTE. Currently, the most promising new therapeutic option is the first of the oral direct thrombin inhibitors, ximelagatran. The THRombin Inhibitor in VEnous thromboembolism (THRIVE) clinical trial programme has demonstrated that this agent is as effective as standard therapy for the acute treatment (THRIVE Treatment) and secondary prevention (THRIVE lll) of VTE events and is well tolerated when used for 6 months or over extended periods up to 1.5 years. Furthermore, with oral administration, fixed dosing and no requirement for anticoagulation monitoring, ximelagatran has the potential to facilitate optimal use and duration of VTE treatment by overcoming the limitations of current agents. PMID- 15812201 TI - Preventing stroke in atrial fibrillation: the SPORTIF programme. AB - Atrial fibrillation (AF) is the most common cardiac risk factor for stroke. Oral anticoagulants such as the vitamin K antagonist warfarin have been proven effective in reducing the risk of stroke in AF. Warfarin, however, has many disadvantages including the need for coagulation monitoring, a narrow therapeutic index, inter-/intra-patient variability and food-drug interactions. As a result, warfarin is underused in clinical practice and a viable alternative is needed. Ximelagatran, the first oral direct thrombin inhibitor, is given as a fixed dose, does not have a narrow therapeutic index, has low potential for drug interactions, has no significant food interactions and does not require coagulation monitoring. Ximelagatran has been evaluated in the Stroke Prevention using an ORal direct Thrombin Inhibitor in atrial Fibrillation (SPORTIF) trial programme, the largest clinical trials of antithrombotic therapy for stroke prevention in AF to date. The phase III trials, SPORTIF III and V, compared ximelagatran (36 mg twice daily) with well-controlled warfarin (international normalized ratio 2.0-3.0) in a combined population of more than 7,000 moderate- to high-risk AF patients. Data from SPORTIF III show an absolute reduction in stroke and systemic embolic events with ximelagatran compared with warfarin at 21 months (1.6 vs. 2.3% per year, respectively; p = 0.10). Preliminary data from SPORTIF V appear to further support non-inferiority between the two agents. On treatment analysis of the rate of major bleeding events shows an absolute, nonsignificant reduction in the event rate per year with ximelagatran versus warfarin in both studies. The results of SPORTIF III and V demonstrate that a fixed oral dose of ximelagatran, without coagulation monitoring, is comparable to dose-adjusted warfarin in preventing stroke and other thromboembolic complications among moderate- to high-risk AF patients and has a lower rate of both major and minor bleeding. With its positive benefit-risk ratio, ximelagatran may increase the population of eligible patients for anticoagulation with AF and maximize the potential of anticoagulation in the prevention of stroke. PMID- 15812202 TI - Cellular and molecular gateways to urolithiasis: a new insight. AB - Urolithiasis is a relevant clinical problem in everyday practice with a subsequent burden for the health system. Urolithiasis is classically explained as the derangement in the process of biomineralization involving the equilibrium between promoters and inhibitors of crystallization: a deficit of one or several inhibitors or an excess of one or several promoters plays a pivotal role in the stone formation. The revolutionary introduction of the molecular biology in medicine has given a new insight in urolithiasis too. Genetic factors have also been postulated to play an important role. A review of the current knowledge on urolithiasis based upon a molecular and genetic approach is reported. PMID- 15812203 TI - Intraindividual variation in total and percent free prostate-specific antigen levels in prostate cancer suspects. AB - OBJECTIVES: To investigate intraindividual total and percent free serum prostate specific antigen (PSA) in prostate cancer suspects and to understand the clinical implications. PATIENTS AND METHODS: Total and percent free PSA were measured using Tandem-R or chemiluminescent enzyme-linked immunoassay for a median of three times in 126 men. Prostate biopsies were performed in all patients; benign prostatic hyperplasia was diagnosed in 81 patients and prostate cancer in 45 patients. RESULTS: The overall mean coefficients of variation for total and percent free PSA were 16.10 +/- 11.94% and 15.45 +/- 15.91%, respectively. A significant correlation (p = 0.0056) was observed between the two variations. The variations in total and percent free PSA were related to none of such stratifications as baseline total PSA level, histology, age, or measurement interval, but for measurement interval on that for total PSA. CONCLUSION: Intraindividual variation in serum PSA should be considered in decision-making about performing prostate biopsies. Also, care should be taken in interpreting repeated percent free PSA measurements in order to enhance the specificity of total PSA, because it had a similar variation to total PSA variation. PMID- 15812204 TI - Optimization of prostatic biopsy: a prospective randomized trial comparing the sextant biopsy with a 10-core biopsy. Impact of prostatic region of sampling. AB - OBJECTIVE: New prostatic biopsy protocols suggest to increase the core numbers to enhance detection. Additional cores are usually sampled from the lateral part of the p-zone. We direct the sextant biopsy to the most lateral part of the p-zone, therefore we investigated if there is a gain by adding 4 median biopsy cores. MATERIAL AND METHODS: The prospective randomized trial (n = 200) compared our modified sextant biopsy to a 10-core strategy with 2 additional median cores on both sides. Directed biopsies to suspicious areas were allowed in both groups. Morbidity was assessed by a self-administered questionnaire. RESULTS: PC detection was 32% for 6 cores and 40% for 10 cores. Four patients were detected only by median biopsies. Using the binomial distribution table the gain of 4% is statistically significant. There was no statistical difference in morbidity, but a trend towards a higher rate of side effects in the 10-core group. CONCLUSIONS: The gain in prostate cancer detection rate by additional median biopsies is low, but statistically significant. There is no difference in morbidity and patient acceptance is high, therefore we favor the 10-core biopsy in our patients. PMID- 15812205 TI - Local anesthesia reduces pain associated with transrectal prostatic biopsy. A prospective randomized study. AB - INTRODUCTION: To test the hypothesis that periprostatic block could completely relief prostatic biopsy-associated pain. MATERIALS AND METHODS: Patients scheduled for transrectal ultrasound guided prostate biopsy were randomized (1:1:1 ratio) to no analgesia (group A), endorectal enema of 1% lidocaine gel (group B) or transrectal periprostatic block (group C). All patients underwent 10 core TRUS-guided biopsy. After the procedure, a ten visual analogue pain score (VAS) from 0 = no discomfort to 10 = severe pain was administered to the biopsied patients and a global estimation of pain associated with the procedure was obtained. The study design included interim analysis of pain score after the first 60 patients were enrolled. Kruskal-Wallis test for unpaired data was used for statistical analysis. Data are presented as mean, median (range). RESULTS: Sixty patients were enrolled between May 2003 and December 2003 and all patients were evaluable. Mean and median age was 68.5 and 69 (range 53-82) years, respectively. Mean and median PSA was 86.8 and 9 ng/ml (range 0.58-4.111), respectively. No major side effects were observed. Patients in group A scored at VAS a median 4, mean 5.5 +/- 2.3 (range 3-10). Patients in group B scored a median 4, mean 5.5 +/- 2.7 (range 3-10) (p = 0.237). Patients receiving periprostatic injections of carbocaine (group C) scored a median 0, mean 0.5 +/- 0.8 (range 0-2). The level of pain reported by this group of patients was significantly different from those reported by patients who performed prostatic biopsy without anesthesia or with intrarectal anesthetic jelly (p = 0.00001). In the periprostatic block group 65% of patients referred no pain after the procedure (VAS = 0) while all patients in the other groups experience some degree of pain. CONCLUSION: The use of bilateral periprostatic block is a very effective and useful technique, well tolerated by the patient, which almost completely abolishes the pain and discomfort associated with the prostatic biopsy procedure. PMID- 15812206 TI - 11C-choline positron emission tomography in prostate cancer: primary staging and recurrent site staging. AB - OBJECTIVES: To evaluate the usefulness of 11C-choline positron emission tomography (PET) for primary staging and re-staging of prostate cancer. PATIENTS AND METHODS: 11C-choline PET, a total of 22 scans, was performed on 13 patients with histologically proven prostate cancer in primary staging (n = 6) and recurrent site staging; following radical prostatectomy (n = 5) and following radiation therapy (n = 3). In 1 patient, 11C-choline PET was performed in both primary staging and re-staging. Also, 3 patients histologically proven to have no malignant prostate were included. RESULTS: Because urinary 11C-choline activity was low, it did not interfere with the visualization of pelvic structures. 11C choline PET visualized normal prostate with a mean SUV of 2.99 (range 2.27-3.68) and primary prostate cancer as a hot spot in 5/6 scans with a mean SUV of 4.21 (range 2.99-6.2). In re-staging, 11C-choline PET was true positive in 9/16 scans and true negative in 2/16 scans. 5/16 scans in 2 patients were false negative with negative conventional imaging. CONCLUSIONS: In primary staging, 11C-choline PET may not be of use because of no reliable differential 11C-choline uptake of BPH and prostate cancer. On the other hand, 11C-choline PET may be of value in recurrent site staging and monitoring for the prostate cancer. PMID- 15812207 TI - Surgical outcome of radical retropubic prostatectomy is not adversely affected by preexisting three-piece inflatable penile implant. AB - OBJECTIVES: We report on our surgical findings and outcome of radical retropubic prostatectomy (RRP) in patients who have had three-piece inflatable penile implants. METHODS: We looked at the retrospective data of 6 patients who have undergone RRP for cancer of the prostate after a prior insertion of a three-piece inflatable penile implant. We looked at the data involving operating time, blood loss, infection of the penile implant, damage to the penile implant, surgical margins of the prostate, damage to other pelvic structures, length of hospital stay, time interval between penile implant and RRP, technique of placing of the reservoir during the penile implant surgery. RESULTS: The RRP procedures all went well, with no injury to the preexisting penile implants. There was also no compromise in the surgery for the cancer treatment, or injury to other pelvic organs. CONCLUSIONS: With increasing treatment of men with erectile dysfunction and also with increasing early diagnosis of cancer of the prostate, we are going to be seeing increasing numbers of patients who have had inflatable penile implants who subsequently need RRP for treatment of cancer of the prostate. We feel the surgery can be accomplished without any complication to the implant or compromise to the cancer surgery or injury to other pelvic organs. PMID- 15812208 TI - Preliminary results of the effect of extracorporeal magnetic stimulation on urinary incontinence after radical prostatectomy: a pilot study. AB - INTRODUCTION: Radical prostatectomy is a common procedure for the treatment of clinically localized prostate cancer. However, urinary incontinence is a significant potential source of morbidity following surgery. Extracorporeal magnetic stimulation (ExMS) is a new technology used for pelvic muscle strengthening in the treatment of stress urinary incontinence. We investigated the clinical effects of ExMS on urinary incontinence after retropubic radical prostatectomy. PATIENTS AND METHODS: Ten patients who had suffered from urinary incontinence for more than 12 months following radical prostatectomy were enrolled in this study. The Neocontrol system was used. Treatment sessions were for 20 min, twice a week for 2 months. The frequency of the pulse field was 10 Hz for 10 min, followed by a second treatment at 50 Hz for 10 min. Objective and subjective measures included voiding diaries, 1-hour pad weight testing, and a quality of life survey at 1, 2, 3, and 6 months after starting the treatment. Urodynamic studies were performed before and after treatment. RESULTS: Three patients became dry (30%), 3 patients improved (30%), and 4 patients showed stationary symptoms (40%). In the 1-hour pad weight testing, the mean pad weight decreased from 25 to 10.3 g, and the quality of life scores had improved from 70.5 to 84.9 2 months after treatment. The frequency of leak episodes per day was reduced from 5.0 times before to 1.9 times after treatment. In the urodynamic study, mean maximum cystometric capacity and Valsalva leak point pressure increased from 197 +/- 53.2 to 309 +/- 85.3 ml and from 67.3 +/- 22.6 to 97.1 +/- 22.7 cm H2O after treatment, respectively (p < 0.05). 3 of 6 patients who showed improvement returned to the baseline values within 12 months after treatment and requested maintenance ExMS therapy. No side effects were observed. CONCLUSIONS: ExMS therapy offered a new option for urinary incontinence treatment after radical prostatectomy. Further studies are required to determine how long the benefits of treatment last and whether maintenance therapy is necessary. PMID- 15812209 TI - Experience with primary urethral carcinoma from the blackfoot disease-endemic area of South Taiwan: increased frequency of bulbomembranous adenocarcinoma? AB - OBJECTIVES: To describe and compare primary urethral carcinomas in South Taiwan with those in the USA and to explore the influence of chronic arsenic exposure. METHODS: From 1988 to 2001, there were 21 pathologically proven primary urethral carcinomas diagnosed and treated at our hospital (14 males, 7 females). Seven of 14 male patients were chronically exposed to arsenic in drinking water for an average of 23 years. We compared our cases to three studies in the USA (80 males, 179 females), and analyzed the influence of chronic arsenic exposure by onset age, histology, staging, and outcome. RESULTS: Male patients with localized tumors had better survival compared to those with advanced tumors (p = 0.0045 in males, p = 0.07 in females). In comparison to the three studies in the USA, there was an unusual higher frequency of bulbomembranous adenocarcinoma at our center (43 vs. 18%, 2 and 0%, respectively, p < 0.0001), particularly among those with chronic arsenic exposure (73 vs. 14%, p = 0.031). CONCLUSIONS: In South Taiwan, there was a high frequency of bulbomembranous urethral adenocarcinoma, which might be associated with chronic arsenic exposure. Although the implications of such an observation are minimal owing to its rarity, it is worth exploring. PMID- 15812210 TI - Diagnosing the location of carcinoma in situ (CIS) of the urinary bladder using pirarubicin hydrochloride. AB - INTRODUCTION: No established technique for locating solitary carcinoma in situ (CIS) of the urinary bladder or CIS accompanying bladder cancer has been determined. Here we investigated whether the location of CIS of the urinary bladder can be macroscopically ascertained by instilling pirarubicin hydrochloride (THP) into the urinary bladder. PATIENTS AND METHODS: We dissolved 50 mg of THP in 50 ml of distilled water, and instilled the resulting solution into the urinary bladder. After 5 min, the urinary bladder is examined using a cystoscope. The study group consisted of 30 subjects (23 men and 7 women). RESULTS: THP uptake was seen in 19 flat (nontumorous) areas of the bladder mucosa in 13 patients. Of these, 11 lesions in 6 patients were confirmed to be CIS. THP uptake was also seen in flat malignant lesions such as bladder cancer invasion into the prostatic urethra, and in benign lesions such as chronic cystitis and urothelial hyperplasia. CONCLUSIONS: The present method can be useful to find easily and macroscopically the location of flat malignant lesions such as CIS. PMID- 15812211 TI - Clinical experience of laparoscopic-assisted radical cystectomy with continent ileal reservoir. AB - INTRODUCTION: Laparoscopic-assisted radical cystectomy with continent ileal reservoir reconstruction is a challenging procedure. We report our experience learned from a hand-assisted laparoscopic technique in our transition to a pure laparoscopic approach. MATERIALS AND METHODS: Eighteen consecutive patients underwent laparoscopic-assisted radical cystectomy with continent ileal reservoir. Hand-assisted laparoscopy was used for the first 11 patients and pure laparoscopy was performed for the subsequent 7 patients in radical cystectomy. Continent ileal reservoir reconstruction was performed extracorporeally via the hand port wound or trocar extension wound. Surgical outcome was analyzed in a retrospective review of the medical records. RESULTS: In the hand-assisted and pure laparoscopic groups, the mean operative time for cystectomy was 2.5 and 2.3 h, for continent ileal reservoir reconstruction it was 4.0 and 3.7 h, and for bilateral lymph node dissection it was 0.8 and 0.6 h, respectively. Mean intraoperative blood loss was 286 ml for the hand-assisted and 179 ml for the pure laparoscopic group. There were no major intraoperative complications nor need for conversion in any of the procedures. Bowel movement was regained in a mean of 3 days and the mean length of postoperative hospital stay was 7 days. CONCLUSIONS: The hand-assisted laparoscopic experience of radical cystectomy learned from the initial 11 patients effectively helped us in the transition to a pure laparoscopic approach. A comparable surgical outcome was found in both groups of patients. PMID- 15812212 TI - Genitourinary cancer: psychological assessment and gender differences. AB - INTRODUCTION: The study of the psychological impact in patients diagnosed with cancer is an important topic. Cancer diagnosis can induce feelings of anxiety, depression, fear of recurrence and produce treatment side effects. Men and women tend to differ in their way of dealing with emotional distress and physical illness. The aim of this study is to investigate and evaluate the prevalence and detection of psychological disease in patients with genitourinary tumors and outline the eventual gender differences. MATERIALS AND METHODS: State-Trait Anxiety Inventory, Self-Rating Depression Scale and Illness Behavior Questionnaire were administered to 60 patients with genitourinary neoplasm. RESULTS: Most patients show symptoms of depression and anxiety during the hospitalization. We have observed a significant gender differences in the psycho emotional assessment. CONCLUSION: Our study confirms the importance to make available for the patients a proper and continuous support in pre- and postsurgery period to prevent the generation of psychological disorders. PMID- 15812213 TI - Effect of urinary leakage on sexual function during sexual intercourse. AB - AIM: This study was planned to determine the effect of urinary leakage on sexual function during sexual intercourse. METHODS: The study group included 32 incontinent women who had urinary leakage during sexual intercourse and the control group consisted of 60 women with no such problem. The Sexual History Form of Schover and Jensen was completed in face-to-face interviews in a private room. RESULTS: When compared with the women without any problem, the women with urinary incontinence were 4.7 times less satisfied with their sexual life and their partners had ejaculation without full erection 3.1 times more. In order to cope with the problem of urinary incontinence during coitus the women themselves adopted several methods. Trying to keep their partners unaware of the problem and deferring the intercourse were among the most frequently adopted methods. 43.7% of the study group indicated that this problem affected their sex life. CONCLUSIONS: The results of this study support the view that urinary leakage during coitus affects women's sex life adversely. PMID- 15812214 TI - Use of alpha1-blockers in female functional bladder neck obstruction. AB - INTRODUCTION: Bladder outflow obstruction may cause obstructive or irritative symptoms. The diagnosis of female functional bladder neck obstruction requires a pressure/flow study and electromyography performed by videourodynamics. The treatment includes self-catheterization or bladder neck incision. We administered tamsulosin, an alpha1A/alpha1D-selective adrenergic antagonist, in women with functional bladder neck obstruction to evaluate its potential therapeutic effects. PATIENTS AND METHODS: A group of 18 women affected by functional bladder neck obstruction was selected. The diagnosis was made by means of a pressure/flow study combined with electromyography and a fluoroscopic test. The diagnostic criteria were: high detrusor pressure with reduced maximum flow, silent electromyography activity, and bladder neck nonfunnelling during the fluoroscopic test. Tamsulosin 0.4 mg once daily was administered for at least 30 days. Patients with a postvoid residual urine volume > or = 100 ml performed intermittent self-catheterization. Patients with a postvoid residual urine volume < 100 ml performed self-catheterization every 7 days. After 30 days of therapy, all patients underwent a new pressure/flow study and a micturition fluoroscopic test. RESULTS: 10 (56%) out of 18 treated patients showed a statistically significant improvement in symptoms, maximum flow, and postvoid residual urine volume (p < 0.01). CONCLUSION: The use of alpha1-blockers may be an initial treatment option for female functional bladder neck obstruction, as this therapeutic option proved to be effective in more than 50% of our patients suffering from this voiding dysfunction. PMID- 15812215 TI - Importance of magnesium in absorption and excretion of oxalate. AB - INTRODUCTION: Magnesium treatment for calcium oxalate urolithiasis is discussed controversially. The aim of this study was to investigate the influence of magnesium supplementation on the oxalate absorption. MATERIALS AND METHODS: The [13C2]oxalate absorption test was always performed three times in 6 healthy volunteers under standardized conditions, with one 10-mmol magnesium supplement together with the labeled oxalate and with two 10-mmol magnesium supplements given in 12-hour intervals. RESULTS: The mean intestinal oxalate absorption under standard conditions was 8.6 +/- 2.83%. The oxalate absorption with one 10-mmol magnesium supplement was 5.2 +/- 1.40% and with two supplements 5.5 +/- 1.62%. Both decreases were statistically significant relative to the standard test, however, not significantly different from each other. CONCLUSIONS: The results show that magnesium administration decreases the oxalate absorption, when magnesium is taken together with oxalate. However, magnesium administration does not decrease the oxalate absorption, when magnesium and oxalate intake differ by 12 h. PMID- 15812216 TI - Glucose transporter-1 expression in urothelial papilloma of the bladder. AB - INTRODUCTION: Urothelial papilloma should be distinguished from low-grade urothelial carcinoma. We report our experience with glucose transporter-1 (Glut 1) expression in urothelial papilloma. MATERIALS AND METHODS: From January 1987 to December 2002, 32 patients with papilloma were diagnosed. Glut-1 protein expression was studied in 32 cases of papilloma and 30 cases of carcinoma. Clinical information was obtained by chart review. RESULTS: Mean age was 54.7 years, and the solitary lesion was 78.1%. No case of bladder papilloma expressed Glut-1 protein even in cases of recurrent papilloma. However, 67% of urothelial carcinoma specimens were stained for Glut-1 protein. Two patients developed recurrent papilloma (6.2%), 1 of whom (3.1%) developed an urothelial carcinoma 4 years after diagnosis of papilloma. CONCLUSIONS: The study showed that papilloma does not express the Glut-1 protein in contrast to urothelial carcinoma. This distinctive expression pattern of Glut-1 will possibly contribute to differentiate urothelial papilloma from low-grade urothelial carcinoma. PMID- 15812217 TI - Short-term effects of arsenic sulfur in deficits of contractile and relaxant responses on urinary bladder: pharmacological and structural changes. AB - INTRODUCTION: Our goal was to investigate the effects of arsenic sulfur (AsS) on the detrusor smooth muscle reactivity. MATERIAL AND METHODS: AsS (100 ppm microg/g) in drinking water was administered for 2 weeks to two groups of female Wistar rats. The contractile responses of urinary bladders to electrical field stimulation, carbachol, ATP and KCl, and the relaxant responses to ATP, adenosine and isoproterenol were examined. Urinary bladder strips were collected for light microscopic examination. RESULTS: Our results demonstrate that oral inorganic AsS exposure induced enhanced contractile and reduced relaxant responses in rats. We hypothesize that AsS is involved in deficiencies of isolated urinary bladder in rats. CONCLUSION: These functional and morphological changes may be important as an intermediate link in urinary bladder oncogenesis induced by AsS. PMID- 15812218 TI - Micturition Syncope due to paraprostatic pheochromocytoma. AB - Paraprostatic pheochromocytoma is a very rare localization of extra-adrenal pheochromocytoma. The clinical and anatomical features of a 41-year-old man are described. PMID- 15812219 TI - Necrotic pancreatic graft causing acute urinary retention. AB - Urologic complications are frequently encountered after pancreas transplantation with a graft duodenocystostomy. Urinary infections, hematuria, and irritative voiding symptoms are some of the most frequent complications with a bladder drained pancreas allograft. While voiding dysfunction is common in these patients due to underlying neuropathy, to our knowledge, we report the first case of obstructive uropathy in a female due to retained necrotic graft material. PMID- 15812220 TI - EBER expression of pure urinary bladder lymphoepithelioma-like carcinoma in two unique Asian patients. AB - We describe the 2 unique Asian cases of pure urinary bladder lymphoepithelioma like carcinoma (LELCA). One patient had metastatic pelvic lymphoadenopathy and the other with superficial tumor. The EBER in situ hybridization both showed negative results despite racial or geographical factors might influence the peculiar susceptibility of individuals of Asian ancestry to Epstein-Barr virus associated lymphoepithelioma-like carcinoma. The 2 patients were treated pertinently with satisfactory outcomes and intravesical BCG instillation is performed to prevent superficial LELCA recurrence rationally. PMID- 15812221 TI - Pure squamous cell carcinoma after intravesical BCG treatment. A case report. AB - Besides the commonly seen side effects of intravesical BCG instillations, very rare complications have been reported including new tumor formation other than transitional cell carcinoma of the bladder. Here we report the first case of pure squamous cell carcinoma formation in a BCG-treated patient with pure transitional cell carcinoma. PMID- 15812222 TI - 'Successful' chemo- and radiotherapy prior to radical cystectomy does not necessarily correlate with clinical course in small cell carcinoma of the bladder. AB - We report 2 cases in which chemo- and radiotherapy induced pathological remission prior to radical cystectomy in small cell carcinoma of the bladder. However, one of these cases suffered a local recurrence 8 months after surgery. PMID- 15812223 TI - Good manufacturing practice production of adenoviral vectors for clinical trials. AB - The increasing importance of recombinant adenoviral vectors for gene therapy, cancer therapy, and the development of prophylactic and therapeutic vaccines has led to worldwide efforts toward scalable process development suitable for commercial manufacturing of replication-deficient adenoviral vectors. This review focuses on the manufacturing of adenovirus for clinical trials in the context of good manufacturing practice conditions and regulations. PMID- 15812224 TI - Protective immunity against alpha-cobratoxin following a single administration of a genetic vaccine encoding a non-toxic cobratoxin variant. AB - Venomous snakebites result in almost 125,000 deaths per year worldwide. We present a new paradigm for the development of vaccines to protect against snakebite, using knowledge of the structure and action of specific toxins combined with a gene-based strategy to deliver a toxin gene modified to render it non-toxic while maintaining its three-dimensional structure and hence its ability to function as an immunogen. As a model for this approach, we developed a genetic vaccine to protect against alpha-cobratoxin (CTX), a potent, post-synaptic neurotoxin that is the major toxic component of the venom of Naja kaouthia, the monocellate cobra. To develop the vaccine, substitutions in the CTX cDNA were introduced at two residues critical for binding to the nicotinic acetylcholine receptor (Asp27 to Arg, Arg33 to Gly). The mutated CTX expression cassette was delivered in the context of a replication deficient adenovirus vector (AdmCTX). To assess whether expression of the mutated CTX in vivo leads to the development of protective immunity, BALB/c mice were challenged by IV administration of 2 microg of alpha-cobratoxin protein 21 or 63 days after administration of AdmCTX or Ad- Null (as a control; both, 10(9) particle units). Animals receiving AdmCTX but no alpha-cobratoxin challenge suffered no ill effects, but > or =80% of naive animals or those receiving the AdNull control vector died within 10 min from the alpha-cobratoxin challenge. In contrast, 100% of animals receiving a single dose of AdmCTX 21 or 63 days prior to alpha-cobratoxin challenge survived. The data demonstrates that an adenovirus-based vaccine can be developed to protect against lethal challenge with a potent snake venom. The effectiveness of this approach might serve as a basis to consider the development of a global public health program to protect those at risk for death by snakebite. PMID- 15812225 TI - Modified infusion procedures affect recombinant adeno-associated virus vector type 2 transduction in the liver. AB - Recombinant adeno-associated virus (rAAV) vectors have therapeutic potential for the treatment of several types of liver diseases including hepato-deficiency disorders. Most of the preclinical and clinical applications involve the use of adeno-associated vector serotype 2 (AAV-2). However, when this vector is delivered at high doses into the portal vein or hepatic artery, a relatively small number of hepatocytes are stably transduced. We elected to determine if the route of vector administration and altering the vascular delivery route within the liver influenced the relative level of transduction. First, we delivered an AAV vector expressing the human factor IX gene from a liver-specific promoter into the hepatic artery, portal vein, or general circulation of rats. Transgene expression was equal with hepatic artery and portal vein infusion, which was higher than vector administered via peripheral venous infusion. Next, we determined how localized perfusion or changing the vector dwell time affected AAV transduction in vivo. To do this, we infused an AAV vector lacking a functional expression and quantified transduction by quantifying the number of double stranded vector DNA genomes. By increasing vector dwell time in the liver to 5 min, vector transduction was enhanced approximately 4- to 5- fold. To establish if gene transduction could be restricted to a specific anatomic location in the liver, we delivered vector into specific liver lobes by clamping the venous inflow to the middle and left liver lobes (noninfused lobes) and infusing vector into the right two liver lobes through the hepatic artery followed by vector circulation between the two right lobes and general circulation for 5 min. With this selective infusion, 40 to 120 times higher vector genome was observed in the perfused lobes than the nonperfused lobes. All the procedures described in this study were performed without detectable liver injury or toxicity. In all, the present study clearly demonstrated that hepatic arterial infusion of rAAV is effective for liver-directed gene therapy and that other parameters related to blood flow can be adjusted to further optimize gene transfer. PMID- 15812226 TI - Low-dose adenoviral immunotherapy of rat hepatocellular carcinoma using single chain interleukin-12. AB - Generation of antitumor immunity by adenoviral gene transfer of interleukin-12 (IL-12) is a very promising concept in cancer gene therapy. Systemically, IL-12 has provoked toxic side effects at therapeutically relevant doses. Native IL-12 lacks effectiveness in clinical trials even when expressed intratumorally from adenoviral vectors. Our strategy was to increase the therapeutic efficacy of IL 12 by expressing a fusion protein of its two subunits (scIL-12) in an adenoviral vector and to evaluate the effects after intratumoral administration. In a rat model of hepatocellular carcinoma, this vector revealed antitumor effects even at a low dosage of 4.6 x 10(5) i.u. in a dose-dependent manner. Long-term antitumor effects were determined at 2.3 x 10(6) and 2.3 x 10(7) i.u. per animal, resulting in 82% and 90% surviving animals, respectively. Magnetic resonance imaging (MRI) enabled individual tumor size follow-up and revealed the scIL-12 effects on large tumors. Treating one hepatic lesion also led to tumor elimination in a second non treated hepatic lesion. Animals rechallenged with tumor cells remained tumor free. Compared to studies applying native IL-12, our data show that the fusion of IL-12 subunits provides approximately 1000-fold higher biological activity. As a consequence of the observed gain in activity, scIL12 promises a substantially improved antitumor efficacy and safety profile of intratumoral adenoviral IL-12 immunotherapy, supporting its clinical use. PMID- 15812227 TI - Adenovirus-mediated transfer and overexpression of heme oxygenase 1 cDNA in lungs attenuates elastase-induced pulmonary emphysema in mice. AB - Heme oxygenase 1 (HO-1) is an inducible enzyme that catalyzes heme to generate bilirubin, ferritin, and carbon monoxide. Because enhanced expression of HO-1 provides an anti-inflammatory effect and confers cytoprotection, we examined whether HO-1 overexpression induced by inoculation of mice with an adenovirus encoding HO-1 (Ad.HO-1) in the lung would prevent pulmonary emphysema induced by porcine pancreatic elastase (PPE). Pretreatment with Ad.HO-1, which upregulated production of HO-1 in the lung, attenuated the PPE-induced increase of neutrophils in bronchoalveolar lavage fluid (BALF) and enlargement of alveoli. It also reduced PPE-induced elevated levels of tumor necrosis factor alpha, interleukin (IL)-6, and keratinocyte-derived chemokine, and increased the level of anti-inflammatory cytokine IL-10 in BALF. These results suggest that Ad.HO-1 induced HO-1 overexpression suppressed PPE-induced emphysema by attenuating neutrophilic inflammation via modulating cytokine and chemokine profiles in mouse lungs. PMID- 15812228 TI - Enhancement of interleukin-12 gene-based tumor immunotherapy by the reduced secretion of p40 subunit and the combination with farnesyltransferase inhibitor. AB - Interleukin-12 (IL-12) gene was shown to produce both IL-12 and p40 subunit. The excess production of the p40 subunit as a natural antagonist of IL-12 is a major obstacle of IL-12 gene-based cancer therapy. We previously reported that IL 12N220L gene, which selectively reduces the secretion of the p40 subunit, induces long-lasting stronger type 1 helper T cells (T(H)1) and cytotoxic T lymphocyte (CTL) immunity in hepatitis C virus (HCV) E2 DNA vaccination model and higher protection from challenge with tumor cells expressing E2 than IL-12 in a prophylactic setting. Here, we demonstrated that intratumoral injection of IL 12N220L-expressing adenovirus showed better tumor growth inhibition and higher survival rate than that of IL-12 or granulocyte macrophage-colony stimulating factor (GM-CSF)-expressing adenovirus in a therapeutic setting. In particular, the mice cured by IL-12N220L treatment were protected against intravenous rechallenge of the same tumor cells better than those by IL-12 treatment. In addition, the enhanced antitumor activity of IL-12N220L was confirmed in B16F10 lung metastasis model, which correlated with the frequency of tumor-specific interferon (IFN)-gamma-secreting cells. When tested in CT26/NP tumor that expresses influenza nucleoprotein (NP) as a tumor antigen, IL-12N220L induced stronger NP-specific T(H)1 and CTL responses than IL-12, particularly at a later time point, indicating the generating long-term tumor-specific memory T-cell responses. Moreover, the potent antitumor effects of IL-12N220L were further augmented by combination with chemotherapy using farnesyltransferase inhibitor (FTI), LB42908. Taken together, our results suggest that IL-12N220L is superior to IL-12 in cancer immunotherapy, which can be further enhanced by combination with chemotherapy. PMID- 15812229 TI - Antibiotic-mediated chemoprotection enhances adaptation of E. coli PNP for herpes simplex virus-based glioma therapy. AB - The E. coli PNP suicide gene sensitizes solid tumors to nucleoside prodrugs, such as 6-methylpurine-2'-deoxyriboside (MeP-dR). In this study using lentiviral, MuLv, and HSV-based gene transfer, we quantified thresholds for inhibition of tumor growth and bystander killing by E. coli PNP and tested the role of intestinal flora in this process. Regressions of human glioma tumors following retroviral transduction exhibited dose dependence on both the level of PNP expression and the dose of MeP-dR administered, including strong tumor inhibition when 90-99% bystander cells comprised the tumor mass. A replication competent, non-neurovirulent herpes simplex virus (HSV) deficient in both copies of the gamma-1 34.5 gene was next engineered to express E. coli PNP under the egr-1 promoter (HSV-PNP). HSV-PNP injected intratumorally (17 million pfu/0.05 ml) in nude mice bearing 300 mg human glioma flank tumors produced a delay in tumor growth (approximately 24 days delay to one doubling). MeP-dR treatment after antibiotic therapy (to eliminate enteric flora encoding PNP enzymes) resulted in antitumor enhancement, with arrest of tumor growth (delay to doubling >50 days). Bystander killing of the magnitude described here has been difficult to accomplish with other suicide genes, such as HSV-tk or cytosine deaminase. The results establish a model for applying E. coli PNP to HSV treatment of glioma. PMID- 15812230 TI - Nonreplicating recombinant vaccinia virus expressing CD40 ligand enhances APC capacity to stimulate specific CD4+ and CD8+ T cell responses. AB - Recombinant poxviruses expressing immunomodulatory molecules together with specific antigens represent powerful vaccines for cancer immunotherapy. Recently, we and others have demonstrated, in vitro and in vivo, that coexpression of CD80 and CD86 costimulatory molecules enhances the immunogenic capacity of a recombinant vaccinia virus (rVV) encoding different tumor-associated antigens. To further investigate the capacity of these vectors to provide ligands for different costimulatory pathways relevant in the generation of T cell responses, we constructed a recombinant virus (rVV) expressing CD40 ligand or CD154 (CD154rVV). Upon binding the CD40 receptor expressed on antigen presenting cells (APC), this molecule, physiologically expressed on activated CD4+ T cells, increases their antigen presentation and immunostimulatory capacities. Therefore, we evaluated the effects of CD154rVV infection on APC activation and its consequences on T cell stimulation. CD154rVV infection of autologous fibroblasts, monocytes, or iDC promoted the expression of a number of cytokines, including GM CSF, TNF-alpha, and IL-15 in iDC. Most importantly, IL-12 p40 gene expression and protein secretion were induced by CD154rVV but not by wild-type VV (WT VV) in either CD14+ cells or iDC, and these effects could be blocked by anti-CD40 monoclonal antibodies. Furthermore, phenotypic characterization of CD154rVV infected iDC revealed enhanced expression of CD83 and CD86 surface markers as compared with wild-type vaccinia virus infection. As expected, VV infection triggered cytokines gene expression in cultures including APC and T cells from VV immune donors. However, cytokine genes typically expressed by T cell receptor triggered T cells such as those encoding IL-2 and IFN-gamma, or T cell proliferation, were detectable to a significantly higher extent in CD154rVV infected cultures, as compared with WT VV. Activation of specific CD8+ T cells was then investigated using MART-1/Melan-A(27-35) epitope as the model of tumor associated antigen (TAA). In the presence of CD154rVV activated APCs, significantly higher numbers of specific cytotoxic CD8+ T cells were detected, as compared with cultures performed in the presence of WT VV or in the absence of virus. Taken together, these data indicate that functional CD154 expression from rVV infected cells promotes APC activation, thereby enhancing antigen-specific T cell generation. Such a recombinant vector might help bypass the requirement for activated helper cells during CTL priming, thus qualifying as a potentially relevant vector in the generation of CD8+ T cell responses in cancer immunotherapy. PMID- 15812231 TI - Liver-targeted gene therapy by SV40-based vectors using the hydrodynamic injection method. AB - Efficient reconstitution of defective genes in hepatocytes could be used to treat various liver and systemic diseases through gene therapy. To explore the potential of SV40-based vectors in liver gene therapy, we constructed SV/luc, an SV40 T-antigen replacement transduction vector, that was propagated on COS and COT cells, which supply the SV40 T-antigen in trans. For liver targeting, BALB/C mice were injected via the tail vein with SV/luc stocks containing 3 x 10(6) to 10(8) transducing units in a volume of 1-2 ml. Luciferase activity was monitored with a light-detection cooled charged-coupled device (CCCD) camera, which enables continuous in vivo measurement of luc expression. The SV40 vector proved to be efficient in gene delivery to the liver, leading to long-term (> or =107 days) transgene expression in hepatocytes. Optimal results were obtained with 3 x 10(6) to 3 x 10(7) transducing units. The hydrodynamic vector delivery method caused transient liver inflammatory changes, with full recovery within days. Low levels of SV40-neutralizing antibodies were detected in the sera of treated mice; however, there was no indication of vector or transgene-specific cellular immune responses. Vectors packaged in vitro, using recombinant capsid proteins and plasmid DNA, were also effective in liver transduction. These results suggest that SV40 vectors may be useful for liver gene therapy. PMID- 15812232 TI - Adrenomedullin gene delivery alleviates hypertension and its secondary injuries of cardiovascular system. AB - Adrenomedullin (AM) is a hypotensive peptide that functions as an important regulator in the cardiovascular and renal systems. The current study explored the potential therapeutic effects of delivering the human AM cDNA via a novel double stranded adeno-associated virus vector (dsAAV) on hypertension and related complications in spontaneously hypertensive rats (SHR). A single dose of dsAAV-AM vector administered by tail vein injection into adult SHR resulted in significant reduction of systolic blood pressure at 2 weeks after gene delivery. This effect was observed through the entire duration of the experiment period (up to 16 weeks). Administration of dsAAV-AM also resulted in a decrease in total urine microalbumin content. Left ventricle and cardiomyocyte hypertrophy, fibrosis in the heart, glomerular sclerosis, and tubular injuries in the kidney were significantly reduced. Moreover, deterioration of hemodynamic variables was prevented in treated rats, as compared with the control groups. We conclude that AAV-mediated AM delivery can render a longterm and stable reduction of hypertension and protect against renal injury and cardiac remodeling in the spontaneously hypertensive rat model. Further preclinical studies are warranted for the development of a gene therapy strategy for human hypertension. PMID- 15812233 TI - Widespread dispersion of adeno-associated virus serotype 1 and adeno-associated virus serotype 6 vectors in the rat central nervous system and in human glioblastoma multiforme xenografts. AB - The transduction patterns of recombinant adeno-associated virus serotype 1 (AAV1) and serotype 6 (AAV6) vectors were assessed in human glioblastoma multiforme (GBM) cell lines, in human GBM biopsy spheroids, and in tumor xenografts growing in nude rat brains. All the cell lines tested (A172, D37, GaMg, HF66, and U373Mg) were found to be permissive to both AAV1 and AAV6 vectors, and thus displayed a transduction pattern similar to AAV2 vectors. For every cell line tested, the transduction efficiency displayed by AAV2 vectors was better than by isogenic and isopromoter AAV1 vectors. Transduction efficiency was dependent on the viral particle number used, suggesting that the receptors for these vectors are widely distributed in GBM tissues. Interestingly, AAV1, AAV2, and AAV6 vectors were able to infect and transduce the same cells when added simultaneously to monolayer cultures. Infection of human GBM biopsy spheroids with AAV1 and AAV6 vectors resulted in transgene expression both at the surface layers and in the core of the spheroids. Following injection of AAV1 and AAV6 vectors into human GBM biopsy xenografts growing in nude rat brains, reporter gene expression was seen both in the periphery as well as in the central regions of the tumors. When injected into the normal rat brain, both AAV1 and AAV6 vectors were found to transduce several central nervous system (CNS) regions. The presented results suggest a potential therapeutic role for AAV1 and AAV6 vectors in gene therapy for GBM and also for other CNS malignancies. PMID- 15812234 TI - Quantification of residual host cell DNA in adenoviral vectors produced on PER.C6 cells. AB - Recombinant adenoviral vectors for gene therapy and vaccination are routinely prepared on cultures of immortalized cells, allowing the production of vector batches of high titer and consistent quality. Quantification of residual DNA from the producing cell line is part of the purity tests for clinical lots. Stringent guidelines stipulate the maximum acceptable level of DNA per dose of vector, and this quantification is therefore a crucial piece of information for researchers and manufacturers alike. In this paper we describe an optimized assay based on real-time polymerase chain reaction (PCR) for the quantification of residual PER.C6 DNA in recombinant adenoviral vectors. In order to reduce the risk of introducing contaminations and to increase the throughput, the assay was designed to require minimum sample handling. Furthermore, DNA extraction from the samples is not necessary, thereby eliminating the need to account for possible sample losses. We also report the results of the assay qualification, demonstrating that the assay is accurate, precise, and sensitive. Finally, we applied the assay successfully to determine the level of host cell DNA in an adenovirus vector produced on PER.C6 cells throughout a standard purification process. Because of its specifications, we anticipate that the assay can have broad applicability to biologics other than adenoviral vectors produced on PER.C6 cells. PMID- 15812237 TI - Synergy between deacetylase inhibitors and IL-1beta in activation of the serum amyloid A2 gene promoter. AB - Butyrate (NaBu) regulates intestinal inflammatory gene expression in part through inhibition of deacetylase activity, but the exact mechanisms involved remain to be determined. In this study, we showed by Northern blot a synergistic induction of the acute phase protein gene SAA2 with a combination of deacetylase inhibitors (Trichostatin A or NaBu) and IL-1beta in the colon carcinoma cell line Caco-2. While the NF-kappa B DNA-binding site was essential for SAA2 regulation by IL 1beta and deacetylase inhibitors, the C/EBP DNA-binding site modulated SAA2 expression levels, as assessed by transient transfection assays and mutagenesis studies. NaBu was sufficient to induce SAA2 expression after transient treatment with IL-1beta and, conversely, IL-1beta induced SAA2 after transient treatment with NaBu. These data suggest that pretreatment with either NaBu or IL-1beta predisposes the SAA2 promoter to further stimulation. Indeed, both NaBu and IL 1beta led to increased recruitment of NF-kappa B p65, C/EBPbeta, and C/EBP delta, and decreased NF-kappa B p50 and C/EBP alpha DNA-binding to the proximal SAA2 promoter, as assessed by chromatin immunoprecipitation assays. Interestingly, while IL-1beta, in contrast to NaBu, induced histone H4 acetylation, addition of IL-1beta and NaBu increased histone H4 acetylation and both C/EBPbeta and NF kappa B p65 DNA-binding. Therefore, these results suggest that NaBu and IL- 1beta mediate SAA2 synergistic induction by establishing and maintaining similar and complementary chromatin modifications and transcription factor recruitment as well. In addition to global effects, NaBu specifically regulate gene expression, as exemplified by SAA2. PMID- 15812238 TI - Genotype-dependent expression of endothelial nitric oxide synthase (eNOS) and its regulatory proteins in cultured endothelial cells. AB - DNA polymorphisms in endothelial nitric oxide synthase (eNOS) gene have been shown to be associated with constitutive eNOS expression and coronary artery disease (CAD). In the present study we explored the hypothesis whether genotype dependent effects can be maintained in vitro during replication, or the effect is conditional on in vivo biological environments. Human umbilical vein endothelial cells (HUVEC) were collected and cultured from 89 normal deliveries of Mexican Americans. The cells were treated with or without cigarette smoking extracts (CSE) and genotypes of eNOS polymorphisms were determined by PCR. We measured the levels of eNOS by ELISA and its binding proteins including heat-shock protein 90 (Hsp-90) and caveolin-1 by Western blotting. The rare C allele for the promoter T786C polymorphism (0.2), and the rare 4 x 27-bp repeat allele in the intron 4 (0.30) were different from those reported in other populations. Yet, the rare T allele in the exon 7 (G894T polymorphism) was similar as others. After four passages in vitro, both the intron 4 and promoter polymorphisms maintained significant effects on eNOS mRNA levels in HUVECs (P < 0.05). However, the effects on eNOS protein and enzyme activity were less consistent. Although primary smokers had significantly lower eNOS protein levels (P < 0.05), the in vitro CSE treatment on cultured HUVECs only resulted in a significant reduction in NO levels as measured by the stable metabolites of nitrite/nitrate (P < 0.001). Neither Hsp-90 nor caveolin-1--important eNOS regulators--appears to mediate the genotypesmoking effects on eNOS expression although HUVECs did produce more Hsp-90 when exposed to CSE. Our study demonstrates that endothelial cells maintain genotype-dependent expression even after the deprivation of in vivo environment. However, the cigarette smoking-genotype interaction may require such in vivo conditions to be manifested. PMID- 15812239 TI - Spatial and temporal changes in the subcellular localization of the nuclear protein-tyrosine kinase, c-Fes. AB - Tyrosine phosphorylation has emerged as a mechanism to control cellular events in the nucleus. The c-Fes protein-tyrosine kinase is an important regulator of cell growth and differentiation in several cell types, and is found in the nucleus of hematopoietic cells. In this study, we showed nuclear localization of c-Fes in both hematopoietic (K562, TF-1, HEL, U937, and HL-60) and nonhematopoietic cell lines (293T, CaOv3, TfxH, MG-63, HeLa, DU-145) by immunofluorescence and confocal microscopy. c-Fes showed striking changes in subcellular localization at specific stages of mitosis. In interphase cells, the intranuclear distribution of c-Fes was diffuse with occasional bright foci. Some c-Fes was present in the cytosol after breakdown of the nuclear membrane, in prometaphase. At prometaphase and metaphase c-Fes was also associated with the chromosomes, in a punctate pattern that partially overlapped with the centromere. Further comparison with proteins that are known components of the kinetochore suggested that some c-Fes protein was located at the centromeric alpha-satellite DNA, between the kinetochores. At anaphase and telophase, c-Fes was entirely cytoplasmic and no protein was found associated with the chromosomes. The timing of c-Fes' appearance at the centromere coincides with the period of kinetochore assembly. These data suggest that c-Fes is recruited to the kinetochore during mitosis. PMID- 15812240 TI - Effect of RNA interference on Gal alpha 1,3 Gal expression in PIEC cells. AB - Xenotransplantation from pigs to human beings is viewed as a potential solution for the acute organ shortage. However, consequent xenorejection induced by Gal alpha 1,3 Gal (a Gal, Gal antigen) prevents xenotransplantation from clinical application. Thus, the most attracting attempt to prevent xenorejection is the elimination of Gal. Our study suggested that compared with the human alpha 1,2 fucosyltransferase (FT) gene and the porcine antisense alpha 1,3 galactosyltransferase gene, sequence-specific siRNA targeting Gal was capable of suppressing Gal expression markedly, and therefore, significantly inhibiting xenoreactivity and the complement activation with human serum in PIEC cells. We also demonstrated the concordant inhibitory effect of siRNA and the human FT gene on Gal and corresponding functions, which implied a practical significance of combined transgenic strategy. The successful application of vector-based dsRNA-GT may extend the list of available modalities in the abrogation of xenorejection in xenotransplantation. PMID- 15812241 TI - Human genome screen to identify the genetic basis of the anti-inflammatory effects of Boswellia in microvascular endothelial cells. AB - Inflammatory disorders represent a substantial health problem. Medicinal plants belonging to the Burseraceae family, including Boswellia, are especially known for their anti-inflammatory properties. The gum resin of Boswellia serrata contains boswellic acids, which inhibit leukotriene biosynthesis. A series of chronic inflammatory diseases are perpetuated by leukotrienes. Although Boswellia extract has proven to be anti-inflammatory in clinical trials, the underlying mechanisms remain to be characterized. TNF alpha represents one of the most widely recognized mediators of inflammation. One mechanism by which TNFalpha causes inflammation is by potently inducing the expression of adhesion molecules such as VCAM-1. We sought to test the genetic basis of the antiinflammatory effects of BE (standardized Boswellia extract, 5-Loxin) in a system of TNF alpha induced gene expression in human microvascular endothelial cells. We conducted the first whole genome screen for TNF alpha- inducible genes in human microvascular cells (HMEC). Acutely, TNF alpha induced 522 genes and downregulated 141 genes in nine out of nine pairwise comparisons. Of the 522 genes induced by TNF alpha in HMEC, 113 genes were clearly sensitive to BE treatment. Such genes directly related to inflammation, cell adhesion, and proteolysis. The robust BE-sensitive candidate genes were then subjected to further processing for the identification of BE-sensitive signaling pathways. The use of resources such as GenMAPP, KEGG, and gene ontology led to the recognition of the primary BE-sensitive TNF alpha-inducible pathways. BE prevented the TNF alpha-induced expression of matrix metalloproteinases. BE also prevented the inducible expression of mediators of apoptosis. Most strikingly, however, TNF alpha-inducible expression of VCAM-1 and ICAM-1 were observed to be sensitive to BE. Realtime PCR studies showed that while TNF alpha potently induced VCAM-1 gene expression, BE completely prevented it. This result confirmed our microarray findings and built a compelling case for the anti-inflammatory property of BE. In an in vivo model of carrageenan-induced rat paw inflammation, we observed a significant antiinflammatory property of BE consistent with our in vitro findings. These findings warrant further research aimed at identifying the signaling mechanisms by which BE exerts its anti-inflammatory effects. PMID- 15812242 TI - DNA shuffling and screening strategies for improving vaccine efficacy. AB - The efficacy of vaccines can be improved by increasing their immunogenicity, broadening their crossprotective range, as well as by developing immunomodulators that can be coadministered with the vaccine antigen. One technology that can be applied to each of these aspects of vaccine development is MolecularBreeding directed molecular evolution. Essentially, this technology is used to evolve genes in vitro through an iterative process consisting of recombinant generation followed by selection of the desired recombinants. We have used DNA shuffling and screening strategies to develop and improve vaccine candidates against several infectious pathogens including Plasmodium falciparum (a common cause of severe and fatal human malaria), dengue virus, encephalitic alphaviruses such as Venezuelan, western and eastern equine encephalitis viruses (VEEV, WEEV, and EEEV, respectively), human immunodeficiency virus-1 (HIV-1), and hepatitis B virus (HBV). By recombining antigen-encoding genes from different serovar isolates, new chimeras are selected for crossreactivity; these vaccine candidates are expected to provide broader crossprotection than vaccines based on a single serovar. Furthermore, the vaccine candidates can be selected for improved immunogenicity, which would also improve their efficacy. In addition to vaccine candidates, we have applied the technology to evolve several immunomodulators that when coadministered with vaccines can improve vaccine efficacy by fine tuning the T cell response. Thus, DNA shuffling and screening technology is a promising strategy to facilitate vaccine efficacy. PMID- 15812243 TI - Frequent 14-3-3 sigma promoter methylation in benign and malignant prostate lesions. AB - 14-3-3Sigma is a putative tumor suppressor gene involved in cell cycle regulation and apoptosis following DNA damage. 14-3-3Sigma loss of expression has been reported is several human cancers, including prostate adenocarcinoma and precursor lesions, and promoter hypermethylation has been proposed as the mechanism underlying gene silencing. Here, we investigate the frequency and extent of 14-3-3sigma promoter methylation in benign and cancerous prostate tissues. We examined tumor tissue from 121 patients with prostate carcinoma (PCa), 39 paired high-grade prostatic intraepithelial neoplasias (HGPIN), 29 patients with benign prostate hyperplasia (BPH), as well as four prostate cancer cell lines using quantitative methylation-specific PCR (QMSP). The percentage of methylated alleles (PMA) was calculated and correlated with clinical and pathological parameters. RT-PCR was performed in the cell lines to assess 14-3 3sigma mRNA expression. PCa, HGPIN, BPH, and cancer cell lines showed ubiquitous 14-3-3sigma promoter methylation. However, the PMA of HGPIN was significantly lower than that of PCa or BPH (P < 0.0001), while PCa and BPH did not significantly differ. The PMA did not correlate with any clinicopathological parameter. All prostate cancer cell lines expressed 14-3-3sigmamRNA. 14-3-3Sigma promoter methylation is a frequent event in prostate tissues and cancer cell lines. Furthermore, there is a progressive accumulation of neoplastic cells with 14-3-3sigma methylated alleles from HGPIN to PCa, suggesting a role for this epigenetic event in prostate carcinogenesis. However, other mechanisms besides promoter methylation might be required for effective 14-3-3sigma downregulation. PMID- 15812244 TI - Effects of intraperitoneal administration of IFN-alpha for one, four, and fourteen days on amino acid levels in various rat brain regions. AB - Interferon-alpha (IFN-alpha) therapy is strongly associated with certain adverse effects, but the pathophysiologic mechanism is unclear. The present study was designed to investigate the influence of peripherally administered IFN- alpha on amino acid levels in the brain. IFN-alpha was administered intraperitoneally (i.p.) once daily to rats, and their brains were extracted 24 h after the last injection. The levels of glutamate, glycine, taurine, gamma-aminobutyric acid (GABA), and arginine in homogenized samples of the frontal cortex, striatum, hippocampus, amygdala, thalamus, hypothalamus, cerebellum, and brainstem were determined. One day of IFN-alpha treatment induced no significant changes in any of these amino acids. After 4 days of injections, glutamate, glycine, taurine, and gamma-aminobutyric acid levels were significantly higher than those in the control frontal cortex, striatum, hippocampus, amygdala, and thalamus. However, most of these amino acids returned to approximately basal levels, or even lower, with 14-day treatment. Our results suggest that daily peripheral administration of IFN-alpha affects the metabolism of amino acids in the brain. Further studies are necessary to determine if these effects of IFN-alpha on cerebral amino acids are involved in the pathophysiology of IFN-alpha-induced depression. PMID- 15812245 TI - Resistance of paramyxoviridae to type I interferon-induced Bos taurus Mx1 dynamin. AB - Typical targets of type I interferon (IFN)-induced antiviral Mx proteins known to date have been shown to share a common profile: single-stranded negative-sense RNA viruses. Among them, human MxA is known to interfere with the replication of measles, human, and bovine parainfluenza-3 viruses (BoPi3V), that is, three members of the Paramyxoviridae family. Recently, bovine Mx1 protein (BoMx1) was included in the group of Mx proteins with authenticated antiviral potential, as it dramatically represses the replication of vesicular stomatitis virus (VSV). As replication in bovine cells of Pi3, respiratory syncytial (RS), and Sendai (Se) viruses, all members of the same family, is known to be reduced on IFN-alpha incorporation into the culture medium, it was hypothesized that the BoMx1 pathway possibly was involved, its antiviral spectrum thus probably extending to Paramyxoviridae. In this study, probing of BoMx1-inhibiting effects was carried out by infecting a transgenic Vero cell line that allows tightly regulated conditional expression of BoMx1 after doxycycline treatment with a wide array of Paramyxoviridae. Expressing and nonexpressing cells displayed similar viability, cytopathic effects (CPEs), and amounts of infectious virus yields, whatever the infecting virus or the multiplicity of infection (moi) imposed. It is, therefore, concluded that BoMx1 does not interfere with Paramyxoviridae. PMID- 15812246 TI - Effects of IFN-alpha on the inflammatory response of swine leukocytes to bacterial endotoxin. AB - Because low-dose interferon-alpha (IFN-alpha) treatment had proved effective in several models of chronic inflammation and autoimmune disease, a possible role of IFN-alpha in modulating the response of swine leukocytes to bacterial endotoxin was investigated in this study. Exposure of swine peripheral blood mononuclear cells (PBMC) to low concentrations of human IFN-alpha caused a strong, dose dependent decrease in CD14 expression, the lowest level being observed at 5 U/ml IFN-alpha. This result was confirmed if PBMC were later exposed to purified lipopolysaccharide (LPS). A 10-fold lower IFN-alpha concentration (0.5 U/ml) caused the largest reduction of tumor necrosis factor-alpha (TNF-alpha) accumulation in the medium of pulmonary alveolar macrophages (PAM), stimulated with bacterial LPS. At 0.5 U/ml, the expression of the TNF-alpha gene in PAM was also strongly reduced, as opposed to cells pretreated with 50 U/ml IFN-alpha. In contrast, expression of the interleukin-1beta (IL- 1beta) gene was stimulated and that of the IL-6 gene was not significantly affected at both IFN-alpha concentrations. Results point to an important role of IFN-alpha in control of the inflammatory response to bacterial endotoxin in pigs. PMID- 15812247 TI - Cholera toxin potentiates influences of IFN-gamma through activation of NF-kappaB and release of tumor necrosis factor-alpha. AB - Cholera toxin (Ctx) is a potent adjuvant in the mucosal immune system. Previous studies have indicated that Ctx induces intestinal interferon-gamma (IFN-gamma) production and that adjuvant properties require activation of the IFN-gamma receptor (IFNGR). Thus, we hypothesized that Ctx potentiates IFN-gamma responses in intestinal epithelia. Initial studies suggested that Ctx enhances IFN-gamma mediated barrier disruption in cultured intestinal epithelia. This response was attributable to liberation of a soluble mediator into conditioned supernatants, subsequently identified as tumor necrosis factor-alpha (TNF-alpha). Extensions of these findings revealed that the Ctx A subunit induces transcriptional activation of proinflammatory genes in addition to TNF-alpha (interleukin-8 [IL- 8], intracellular adhesion molecule-1 [ICAM-1], and IL-6) and that such transactivation is mediated by the transcriptional regulator NF-kappaB. We conclude that Ctx elicits a proinflammatory phenotype in intestinal epithelia and that potentiation of IFN-gamma-mediated barrier disruption by TNF-alpha may contribute to the overall adjuvant properties of Ctx. PMID- 15812248 TI - Thrombopoietin in the cerebrospinal fluid of patients with aseptic and bacterial meningitis. AB - Despite the recent evidence of the localization of thrombopoietin (TPO) and its receptor in the central nervous system (CNS), TPO protein concentrations in the cerebrospinal fluid (CSF) remained to be clarified. We previously reported that serum TPO is increased in children with meningitis. To determine changes in TPO concentrations in the CSF by meningitis and to explore the relationship between serum and CSF TPO concentrations, we measured TPO concentrations in 110 CSF samples and 33 serum/CSF pairs from 11 bacterial meningitis, 49 aseptic meningitis, and 50 nonmeningitis children. In only 12% (13 of 110) of CSF samples (0 bacterial meningitis, 8 aseptic meningitis, and 5 controls), TPO concentrations could be determined (24.1 +/- 29.0 pg/ml). CSF TPO concentrations did not significantly differ among the three groups and did not correlate with age. TPO concentrations in all serum samples were detectable, and mean concentrations in bacterial meningitis (510.6 +/- 237.0 pg/ml) were significantly higher than those in aseptic meningitis (136.6 +/- 71.6, p < 0.01) and controls (181.3 +/- 88.3, p < 0.01). These findings suggest that TPO is not produced in the CNS of patients with meningitis and that TPO did not cross the blood-brain barrier even during meningeal infection. PMID- 15812249 TI - Identification of genes differentially expressed in osteoclast-like cells. AB - Homeostasis of the skeletal system is maintained by a balance between bone formation and resorption. The receptor activator of NF-kappaB ligand (RANKL) induces the differentiation of bone-resorbing cells, osteoclasts. To identify genes regulated during osteoclast differentiation, we constructed a subtraction cDNA library using a mouse RAW264 macrophage cell line that differentiates into osteoclast-like multinucleated cells after treatment with RANKL. Northern blot analysis showed that RANKL treatment upregulated expression of 17 genes. Among these were the genes for five H(+)-ATPase subunits, two chemokines, and the osteoclast marker cathepsin K. In addition, a mouse homolog of human dendritic cell (DC)-specific transmembrane protein (DCSTAMP), whose function in osteoclastogenesis was recently revealed, was also included in the induced genes. Characterization of these inducible genes will provide an insight into the biology of osteoclasts and the mechanism of bone-related diseases. PMID- 15812251 TI - Sequential follow-up clinic and ambulatory blood pressure evaluation in a low risk population of white-coat hypertensive patients and in normotensives. AB - BACKGROUND: In subjects with white-coat hypertension (WCH) it is unclear how ambulatory blood pressure (ABPM) progresses over time and whether they exhibit an increased cardiovascular risk. METHODS: We prospectively evaluated the transition of clinic and ABPM values in 39 clinic and ABPM normotensive subjects (NT) (clinic BP<140/90 mmHg and awake BP<130/85 mmHg, ages 43.4+/-2.6 y) and in 79 untreated subjects (47.2+/-2.4 y) with WCH (clinic BP>140/90, awake ABP<130/85 mmHg) with no other major cardiovascular risk factors. Ambulatory blood pressure was evaluated at baseline and on at least two further occasions during follow-up. RESULTS: At baseline all subjects were untreated and groups did not differ on values of metabolic parameters, BMI, left ventricular mass index, and ABPM values. Subjects were revaluated for ABPM half way through and at the end of follow-up, 35+/-3 and 86+/-4 months in NT and 49+/-4 and 90+/-4 months in WCH. Thirty-six WCH were on antihypertensive treatment (AH) after baseline until the end of follow-up (WCH-tr), whereas 43 WCH (WCH-untr) were free from AH throughout the study. In a similar way all groups showed a significant (p<0.01) progressive increase in 24-h ABPM systolic blood pressure (SBP)/diastolic blood pressure (DBP) from baseline throughout the follow-up in NT (+4.9/2.1+/-0.8/0.9 mmHg), average annual increase of 0.72/0.37 mmHg/y, in WCH-tr (+ 5.0/1.2+/-1.1/1.5 mmHg), average annual increase of 0.66/0.31 mmHg/y and in WCH-untr (+5.4/3.2+/ 0.9/1.1 mmHg), average annual increase of 0.74/0.39 mmHg/y. During the follow-up office SBP/DBP (mmHg) significantly rose in NT (+5.7/3.9) but was reduced in WCH tr (-7.8/5.2) and in WCH-untr (-4.7/1.1). Development of ambulatory hypertension (daytime BP >130 and/or >85 mmHg) occurred in 15.4% (6/39) of NT, in 22.7% (8/36) of WCH-tr and in 26.1% (11/43) of WCH-untr (NS). First cardiovascular events recorded were three in subjects with WCH and none in NT. CONCLUSIONS: After 7.4 years of follow-up, both the progressive increase in ABPM and the rate of transition to ambulatory hypertension in subjects with WCH (either treated or untreated), who were selected under strict criteria were similar to that of normotensive subjects. Also there was no evidence that WCH exhibited a clear higher risk of development cardiovascular events. PMID- 15812252 TI - The natural history of white-coat hypertension in the long term. PMID- 15812253 TI - Inaccuracy of wrist-cuff oscillometric blood pressure devices: an arm position artefact? AB - BACKGROUND: Despite the increasing popularity of wrist-cuff blood pressure (BP) devices, their accuracy has not been established and international guidelines do not support their use. Because arm position influences BP measurement, it is possible that conflicting reports on wrist-cuff device accuracy reflects diverse arm positions. METHOD: This study compared BP measured by two oscillometric devices, the upper arm-cuff OMRON HEM 705 CP and the OMRON R6 oscillometric wrist cuff device. In the former BP was measured with the arm in two supported positions, dependent on a table (manufacturer's instructions) and horizontal (mid sternum), while the latter followed the manufacturer's instructions. RESULTS: In contrast to the dependent arm where BP was significantly higher (P<0.05), the horizontal arm position with the arm-cuff produced a mean systolic and diastolic BP comparable to the wrist-cuff device where the wrist was at heart level being respectively, 137+/-29/80+/-16 and 134+/-27/77+/-16 mmHg. A close relationship over a wide BP range was also confirmed by least squares, least product linear regression and Bland-Altman analysis. CONCLUSION: This study supports the use of wrist-cuff monitors for self/home use and underlines the need for a more precise definition for arm position when using all BP devices -- mercury and oscillometric. PMID- 15812254 TI - Arm circumference in adults in Poland as an important factor influencing the accuracy of blood pressure readings. AB - AIMS: The aim of this study was to assess the distribution of right arm circumference (RAC) in adults in Poland and to establish how often other than standard cuffs should be used in everyday practice. We also wanted to examine whether large cuffs are available in the market in Poland and whether the cuffs comply with WHO and JNC VII recommendations. MATERIAL AND METHODS: In 2002, we conducted a cross-sectional representative survey NATPOL PLUS to examine prevalence and control of cardiovascular risk factors in Poland. The survey included a questionnaire interview, blood pressure (BP) and anthropometric measurements, as well as laboratory tests. The examined representative sample of adults in Poland consisted of 3051 men and women aged 18-94 years. The measurements of RAC were performed in all respondents. Arterial hypertension (AH) diagnosis was based on three separate visits (BP > or =140/90 mmHg or treatment). Subjects with body mass index (BMI) > or =25 kg/m were included to overweight/obese group. RESULTS: The prevalence of AH was 29% and of overweight or obesity, 52%. In 24% of all adults RAC was > or = 32 cm, and in 6% was < 24 cm. In hypertensives, the respective figures were 36 and 3%; and in overweight/obese subjects 40 and 0.6%. Mean RAC was 29.2+/-3.8 cm (males 30.2+/ 3.5 cm; females 28.3+/-3.9 cm, P<0.001). Mean RAC was significantly greater (P<0.01) in patients with AH (30.4+/-3.9 cm; males 30.7+/-3.5 cm; females 30.2+/ 3.8 cm) than in normotensive subjects (28.4+/-3.7 cm; males 29.9+/-3.4 cm; females 27.3+/-3.5 cm). However, these differences were not statistically significant when the results were adjusted for age and BMI. Additional analysis of sphygmomanometers available in Poland showed that parameters of standard cuffs fulfilled recommendations of JNC VII and WHO/ISH guidelines. However, the sets with large cuffs were usually much more expensive and only one manufacturer made them available as a special order. CONCLUSIONS: One-third of adults in Poland have an arm circumference, which requires use of a 'non-standard' sized cuff. Moreover, in patients with AH or overweight/obese subjects, wider or narrower cuffs should be used in approximately 40% of subjects. PMID- 15812255 TI - Blood pressure dipping is reproducible in clinical practice. AB - OBJECTIVE: Blood pressure dipping pattern has clinical and prognostic consequences. However, reproducibility of night-time blood pressure fall during 24-h ambulatory blood pressure monitoring is considered limited. This limited reproducibility is possibly a result of inadequate day-night definitions. We retrospectively examined the reproducibility of blood pressure dipping in clinical practice, applying a method that accounts for sleep-awake states and does not rely on arbitrary day-night definitions. We also examined dipping repeatability in subjects with changing blood pressure. METHODS: Of 962 consecutive ambulatory measurements performed in our unit during a 3-year period, 100 patients (age 60+/-15) had a prior session, and were the subjects of this study. Based on patients' report we defined 'awake blood pressure' as the average of pressure recordings while the subject was awake, including night-time arousals, and 'sleep blood pressure' as the average of pressure recordings while the subject was sleeping, including afternoon naps. RESULTS: We found systolic blood pressure dipping not less reproducible than 24-h, awake- and sleep systolic blood pressure, as evaluated by both Pearson correlations (r=0.52 versus 0.5, 0.5, 0.49, respectively, P < or =0.0002 in all), and Bland-Altman repeatability. In a subgroup of 35 subjects (age 63+/-15) with at least 10 mmHg change in systolic blood pressure between the two sessions, systolic blood pressure dipping remained reproducible (r=0.45, P<0.007). CONCLUSIONS: When interpreted in a way that accounts for sleep-awake pattern, sleep-induced systolic blood pressure dipping in clinical practice is a very reproducible feature of ambulatory blood pressure monitoring, in accordance with its vital prognostic implications. PMID- 15812256 TI - 24-hour ambulatory blood-pressure effects of valsartan and hydrochlorothiazide combinations compared with amlodipine in hypertensive patients at increased cardiovascular risk: a VAST sub-study. AB - BACKGROUND: There is a lack of data on the effects of angiotensin-receptor blocker and diuretic combinations on ambulatory blood pressure (ABP) in hypertensive patients with additional cardiovascular risk factors. METHODS: In a randomized, double-blind trial, the effects on 24-h ABP of the combination valsartan 160 mg od and hydrochlorothiazide 25 or 12.5 mg during 24 weeks of therapy were compared with the effects of amlodipine 10 mg monotherapy (group A10) in 474 stage-II hypertensive patients with additional cardiovascular risk factors. After a two-week single-blind placebo run-in period, patients were randomized to receive valsartan 160 mg od or amlodipine 5 mg od. At week 4, HCTZ 12.5 mg (group V160/HCTZ12.5) and 25 mg (group V160/HCTZ25) were added to the valsartan groups and in the A10 patients the amlodipine dose was force-titrated to 10 mg od. RESULTS: All three treatments reduced 24-h BP as well as night-time and daytime BP levels from baseline. Twenty-four hour systolic blood pressure (SBP) was reduced by 15.9+/-1.0 mmHg (least-squares mean change+/-SE), 19.3+/-1.0 mmHg and 16.1+/-1.1 mmHg in the V160/HCTZ12.5, V160/HCTZ25 and A10 groups, respectively and 24-h diastolic blood pressure (DBP) was reduced by 9.3+/-0.6 mmHg, 11.4+/-0.6 mmHg and 9.6+/-0.7 mmHg in the three groups. The differences between the V160/HCTZ25 group and the A10 group were significant (p<0.05) for the changes in 24-h systolic BP as well as for changes in daytime systolic BP and night-time diastolic BP. Control rates defined as ABPM < or =130/80 mmHg were: 48.4%, 60.8% and 50.9% in the V160/HCTZ12.5, V160/25 and A10 groups, respectively. The differences in control rates between the V160/HCTZ25 group and the other two treatment groups were significant at p<0.05. CONCLUSIONS: The fixed dose combination of valsartan 160 mg+HCTZ 25 mg od is an attractive therapeutic option measured on the effects on 24-h ABPM, night-time and daytime BP reduction and control rates in hypertensive patients at additional cardiovascular risk. PMID- 15812257 TI - Ambulatory blood pressure variations relative to sitting or standing position in renal transplant patients. AB - OBJECTIVE: High blood pressure is almost constant in renal transplant patients for whom dysautonomia is frequently described. The main objective of this study was to analyse the variations in blood pressure and heart rate recorded by ambulatory measurement during changes in position in renal transplant patients. METHODS: Thirty-nine non-diabetic renal transplant patients with a renal transplant functioning for more than a year, were selected at random. Blood pressure was measured using the validated monitor Diasys Integra with a position sensor to discriminate between standing and sitting/lying. RESULTS: Systolic blood pressure and heart rate were significantly higher when the patient was standing than when sitting/lying (+2.9 mmHg, P<0.05 and +9 beats/min, P<0.001 respectively) and diastolic blood pressure tends to be higher (+1.7 mmHg, NS) when standing. One minute after standing up, the heart rate rises by about 9 beats/min (P<0.001) while systolic and diastolic blood pressures do not vary significantly. Variations in systolic blood pressure and heart rate on changing position are therefore in the same direction as those recorded in elderly normotensive or hypertensive untreated subjects, but with a lower amplitude. CONCLUSIONS: In most of non-diabetic functional renal transplant patients, there is an absence of an orthostatic decline in blood pressure. Thus, it could be considered that there is no real dysautonomia in this specific population. PMID- 15812258 TI - Validation of the AGILIS ambulatory blood pressure monitor according to the European Society of Hypertension International Protocol for validation of blood pressure measuring devices in adults. AB - BACKGROUND: We evaluated the clinical validity of the AGILIS ambulatory blood pressure monitor according to the European Society of Hypertension International Protocol. METHODS: The blood pressure (BP) was measured at the same arm by two certified observers using two mercury columns and a double stethoscope, then by the volunteer with the automatic device, these measurements were done seven times sequentially using 33 volunteers. For the randomization of the patient, one ABPM measure was compared to the mean of three manual measures. Then, we had three ABPM measures and four manual measures. Each ABPM measure, being compared to two manual measures. Blood pressure readings and recordings were blinded. Position of the subjects was standardized. RESULTS: Phase 1: To pass phase 1, the device had to have at least 25 of the 45 measurements within 5 mmHg, 35 within 10 mmHg or 40 within 15 mmHg of the comparative observer measurements. The AGILIS monitor had 35 measurements lying within 5 mmHg, 42 within 10 mmHg and 45 within 15 mmHg for systolic BP (SBP), 35 within 5 mmHg, 44 within 10 mmHg and 45 within 15 mmHg for diastolic BP (DBP). It passed the criteria for SBP and DBP. Phase 2: To pass phase 2.1, a device must have at least 60 of 99 measurements within 5 mmHg, 75 within 10 mmHg and 90 within 15 mmHg of the comparative observer measurements, and in addition must also have either 65 within 5 mmHg and 80 within 10 mmHg, or 65 within 5 mmHg and 95 within 15 mmHg, or 80 within 10 mmHg and 95 within 15 mmHg. The AGILIS monitor had 78 measurements within 5 mmHg, 91 within 10 mmHg and 96 within 15 mmHg for SBP; and 70 measurements within 5 mmHg, 92 within 10 mmHg and 96 within 15 mmHg for DBP. It passed the criteria for SBP and DBP. For phase 2.2 using the AGILIS monitor, 27 subjects had at least two of the differences within 5 mmHg and none had any differences within 5 mmHg for SBP; 27 subjects had at least two of the differences within 5 mmHg and three subjects had no differences within 5 mmHg for DBP. The results are in concordance with the requested criteria for phase 2.2. CONCLUSION: Thus the AGILIS device fulfils the validation criteria of the international protocol. PMID- 15812259 TI - A survey of validated automated home blood pressure monitors available for the Internet shopper. AB - OBJECTIVE: Self-measurement of blood pressure using an automated home blood pressure monitoring (AHBPM) is increasingly used in hypertension management. Internet commerce increases dramatically each year. This study looked to identify the availability of validity of AHBPM and the correct cuff size to go with the AHBPM. METHODS AND RESULTS: Using the search engine 'Google.com', the author identified 124 consecutive unique sites offering at least one AHBPM. Validated AHBPM were those devices that had published studies showing that they had passed a recognized validation protocol. Each site was evaluated for all forms of sphygmomanometer, number of AHBPM, manual blood pressure devices, all cuff sizes available, additional cost of large adult cuff, number of validated AHBPM offered, and whether the site mentioned device validation. Of the 124 sites, 109 (81%) offered arm AHBPM and 66 (53%) offered one or more (range, 1-11) validated AHBPM. Only six of the 66 (9%) offering a validated AHBPM mentioned that fact; 58 of the 109 (53%) sites offering arm AHBPM offered more than one size of cuff; and 46 of the 58 (80%) charged extra for a large adult cuff (average US dollars 23.75, range, US dollars 4.80-98). CONCLUSIONS: Validated AHBPMs are readily available on the Internet. Currently, these sites do little to aid the consumer in purchasing a validated AHBPM. Large adult cuffs, commonly needed by hypertensive patients, are not always available for purchase. Charging extra for large adult cuffs is a potential hindrance to consumers purchasing the correct cuff size for accurate blood pressure measurement and should be eliminated. PMID- 15812260 TI - Taking blood pressure -- no laughing matter! AB - BACKGROUND: Humour is used commonly to relax subjects when their blood pressure is being measured. However the short-term effect of laughter on blood pressure is not described. METHODS: Sixteen normotensive volunteers had blood pressure measured using a validated automated oscillometric sphygmomanometric device (Omron 705 CP) as control and during three episodes of laughter while viewing a humorous sit-com video. RESULTS: Blood pressure was stable in the control period. There was a significant rise (P<0.01) in systolic blood pressure from 115 mmHg (108-121, mean 95% confidence interval) to 127 mmHg (120-135) during laughing. The increase in diastolic pressure 71 (67-74) to 73 (69-77) mmHg was not significant. CONCLUSION: Laughing has an acute effect on systolic blood pressure. Patients should not be encouraged to laugh when their blood pressure is being measured. PMID- 15812261 TI - Efficacy and safety of direct transition to risperidone long-acting injectable in patients treated with various antipsychotic therapies. AB - The maintained antipsychotic efficacy of risperidone long-acting injectable (RLAI) was investigated in patients with schizophrenia or other psychoses who were transitioned directly from their previous antipsychotic medication. Patients symptomatically stable, but considered to require a treatment change, received 25 mg of RLAI (increased to 37.5 or 50 mg, if necessary) every 2 weeks for 6 months. Assessments included Positive and Negative Syndrome Scale (PANSS), Clinical Global Impression-Severity (CGI-S), Global Assessment of Functioning (GAF), SF-36 Health-Related Quality of Life Questionnaire and Extrapyramidal Symptoms Rating Scale (ESRS). Of 1876 patients enrolled, 74% completed the 6-month study. The most frequent reasons for treatment change were non-compliance (38%), insufficient efficacy (33%) and side-effects (26%). There was a significant reduction from baseline to endpoint in mean total PANSS score and in the scores on all PANSS subscales and symptom factors (P<0.001). CGI-S improved significantly, as did mean GAF score, all factors on the SF-36 and patient satisfaction with treatment. Scores on ESRS showed significant, sustained improvements throughout the study period. Direct initiation of RLAI was effective and well tolerated. RLAI provides an advancement in the treatment options available for a wide range of patients requiring long-term antipsychotic therapy. PMID- 15812262 TI - Prospective, multicentre, randomized, double-blind study of the efficacy of escitalopram versus citalopram in outpatient treatment of major depressive disorder. AB - Pre-clinical studies, active-control clinical trials and meta-analyses indicate that escitalopram (S-citalopram) might be more effective than citalopram, the racemic mixture of S- and R-citalopram. The present study aimed to confirm the superior efficacy of escitalopram over citalopram. A double-blind, randomized clinical trial was performed in which general practitioners and psychiatrists compared fixed doses of escitalopram (20 mg/day) with citalopram (40 mg/day) over 8 weeks in outpatients with major depressive disorder (MDD) [baseline Montgomery Asberg Depression Rating Scale (MADRS) score > or =30]. Primary efficacy parameter was change from baseline to last assessment in the MADRS total score. Out of 138 (aged 44.1+/-10.9 years; initial MADRS score 36.3+/-4.8) and 142 (aged 46.2+/-11.1 years; initial MADRS score 35.7+/-4.4) evaluable patients who were randomized to escitalopram and citalopram, respectively, six and 15 withdrew prematurely (P=0.05). The MADRS score decreased more in the escitalopram than in the citalopram arm (-22.4+/-12.9 versus -20.3+/-12.7; P<0.05). There were more treatment responders with escitalopram (76.1%) than with citalopram (61.3%, P<0.01). Adjusted remitter rates were 56.1% and 43.6%, respectively (P<0.05). Tolerability was similar in both groups. This randomized double-blind trial confirms that escitalopram has a superior effect to citalopram in MDD. PMID- 15812263 TI - Escitalopram and suicidality in adult depression and anxiety. AB - The escitalopram clinical trial database, consisting of all placebo-controlled and relapse prevention trials within major depressive disorder (MDD) and anxiety disorders, was analysed for specific adverse events indicative of suicidal behaviour (fatal suicide, non-fatal self-harm or suicidal thoughts) in relation to treatment. The number of events was low, with no fatal suicides in the first 2 weeks of treatment. There was one fatal suicide during the full treatment period on placebo (incidence 0.1%; rate 0.003), and none on escitalopram. None of these figures were significantly different between escitalopram (n=2277) and placebo (n=1814) patients. There was no indication that escitalopram provokes suicidal behaviour compared to placebo in either MDD or anxiety disorders. Based on efficacy ratings (Montgomery-Asberg Depression Rating Scale, item 10), escitalopram was more efficacious versus placebo in lowering suicidal thoughts from weeks 1 through 8 in the treatment of patients with MDD. PMID- 15812264 TI - The influence of psychotropic drugs on cerebral cell death: female neurovulnerability to antipsychotics. AB - Tissue transglutaminase (tTG) is a marker for apoptosis, and its protein level is known to be increased in post-mortem Alzheimer's and Huntington's disease brains. tTG is increased in the cerebrospinal fluid of patients with Alzheimer's disease. However, the influence of psychotropic medication on acute cell death has not been studied so far in vivo, although some experiments performed in vitro suggest that antipsychotic drugs are neurotoxic. The protein level of tTG was examined in the cerebrospinal fluid obtained from 29 patients under neuroleptic medication in the last 24 h before lumbar puncture (eight patients diagnosed with Alzheimer's disease and 21 patients with other neurological diseases), and compared with those from 55 patients without antipsychotic medication (25 Alzheimer's patients and 30 others). In addition, the influence of several other psychotropic drugs on apoptosis was analysed. A significant influence (P<0.01) of antipsychotic drugs for both the Alzheimer's and the non-Alzheimer's group was found with respect to tTG protein levels in cerebrospinal fluid. By contrast to the male subgroups, the female groups showed a strong influence of neuroleptics on cerebral cell death. Surprisingly, atypical antipsychotics did not differ from typical neuroleptics in neurotoxicity. By contrast, no influence of antidepressants, cholinesterase inhibitors, nootropics, tranquilizers and tramadol on cerebral cell death was found. The results suggest that typical and atypical antipsychotic drugs may induce cerebral cell death, especially in female patients. Subjects with Alzheimer's disease might be even more vulnerable to any antipsychotic. Therefore, subsequent research should aim to identify atypical neuroleptics without neurotoxicity. A limit on the use of first- and second-generation antipsychotics in elderly patients is proposed. Finally, the possible connection between the observed increased cerebral cell death and tardive dyskinesia, the most threatening side-effect in antipsychotic therapy, is discussed. PMID- 15812266 TI - Reduction in hospital stay of chronic schizophrenic patients after long-term clozapine treatment. AB - The present study aimed to elucidate the effectiveness of clozapine treatment in reducing the disabling period of chronically ill schizophrenic patients by investigating their rehospitalization status. Of 232 schizophrenic patients with a history of clozapine use who were recruited from the clinic at Seoul National University Hospital, 117 were selected who had been followed up for more than 1 year with respect to rehospitalization. To obtain information about the period before the clozapine change, a chart review of these 117 patients was conducted. The number and length of hospitalizations of the patients significantly decreased after clozapine treatment compared to the same period before clozapine treatment. The hospital days per year of the patients were also decreased significantly after clozapine introduction. By analysing 38 patients who were followed up for more than 5 years, it was suggested that the decrease in the number and length of hospitalizations was substantially sustained for up to 5 years after clozapine treatment. This study showed that the number and length of hospitalizations are significantly decreased by long-term clozapine treatment and that this effect can positively affect the social outcome of schizophrenic patients. PMID- 15812265 TI - Controlled clinical comparison of paroxetine and fluvoxamine considering the serotonin transporter promoter polymorphism. AB - The present study aimed to compare the effects of two currently used selective serotonin reuptake inhibitors (SSRIs) in Japan taking the individual background in 5-HTT gene-linked polymorphic region (5HTTLPR) genotype into account. Clinical responses to paroxetine and fluvoxamine were evaluated by total and cluster depressive symptoms for 81 Japanese patients who were diagnosed with major depression. Patients with the l allele had a greater percentage reduction on the total score (P=0.059) and somatic anxiety items (P=0.026) of the 21-item Hamilton Depression Rating Scale (HAM-D) score compared to s/s genotype carriers. Paroxetine was significantly more effective than fluvoxamine in the s/s carriers, as evaluated on the percentage reduction in total score (P=0.012) and core (P=0.049) HAM-D after 4 weeks of medication, but not in the l/s carriers. These findings suggest that the genetic test may be useful in investigating the efficacy of the two SSRIs, and that normalization by the 5HTTLPR genotypes may lead to improvement of the precision of comparative analysis. PMID- 15812267 TI - An inter-ethnic comparison study of clozapine dosage, clinical response and plasma levels. AB - The present study investigated clozapine dosage, plasma clozapine and metabolite levels, clinical and side-effect profiles in Asian versus Caucasian patients with chronic schizophrenia who were on stable maintenance treatment. Twenty Asian patients from Singapore and 20 Caucasian patients from Australia were systematically evaluated with the following rating scales: Positive and Negative Syndrome Scale for Schizophrenia, drug attitude scale (DAI-10), drug adverse reaction profile (Liverpool University Neuroleptic Side-effect Rating Scale), extrapyramidal side-effects scales (Abnormal Involuntary Movement Scale, Simpson and Angus Scale). Cigarette and caffeine consumption were recorded and steady state plasma clozapine and metabolites levels were measured. Although Asian patients received a significantly lower mean clozapine dose (176 mg/day) than the Caucasian group (433 mg/day, P<0.001), plasma clozapine levels were similar between the groups. As a result, Asian patients had more than twice the effective clozapine concentration to dose ratio (P<0.001). The findings remained significant even after controlling for gender, body mass index, cigarette, alcohol and caffeine use. Conversely, the plasma metabolites (desmethylclozapine and clozapine N-oxide) to clozapine ratios were higher in the Caucasian patients (P<0.01). Compared to Caucasian patients, Asian patients appeared to have a lower dosage requirement for clinical efficacy. Hence, appropriate dose adjustment should be considered in Asian patients receiving maintenance clozapine therapy in clinical practice. PMID- 15812268 TI - Past use of selective serotonin reuptake inhibitors and the risk of cerebrovascular events in the elderly. AB - Treatment with selective serotonin reuptake inhibitors (SSRIs) has been associated with intracranial bleeding abnormalities. We investigated the relationship between past exposure to SSRIs and occurrence of cerebrovascular accidents in the elderly. From the regional database of hospital admissions of Lombardy, Italy, we extracted all patients aged 65 years or above with cerebrovascular-related outcomes for the year 2002. From the regional database of prescriptions reimbursed by the National Health Service, we extracted all patients aged 65 years or above who received antidepressant prescriptions during 2001. The two databases were linked anonymously using the individual patient code. The analysis showed that the proportion of cerebrovascular events in those exposed to SSRIs was 135/66 335 [0.20%, 95% confidence interval (CI) 0.17-0.24], whereas the proportion of cerebrovascular accidents in those exposed to tricyclic antidepressants (TCAs) was 28/18 620 (0.15%, 95% CI 0.09-0.21). After background group differences were controlled for, exposure to SSRIs did not increase the risk of accidents (adjusted odds ratio 1.31, 95% CI 0.87-1.97). Although the risk of cerebrovascular accidents is a rare but serious event that should carefully be monitored during antidepressant therapy, this study indicated an absence of difference between TCAs and SSRIs. PMID- 15812269 TI - Naturalistic observation on the hepatic enzyme changes in patients treated with either risperidone or olanzapine alone. AB - This retrospective study aimed to compare differences in hepatic enzyme elevation during treatment with either risperidone or olanzapine alone in patients with psychotic disorders. The charts were reviewed for six hundred and sixty-seven (667) inpatients with psychotic disorders who were treated with either risperidone (n=289) or olanzapine (n=145) alone at a university-affiliated hospital between 1998 and 2002. Frequencies of elevation greater than the reference level in any enzyme among aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphotase (ALP) were higher in the olanzapine-treated group (26.9%) than in the risperidone-treated group (14.2%) [odds ratio (OR)=2.225, 95% confidence interval (CI)=1.362-3.638, P=0.002]. Frequencies of elevation greater than the reference level in ALT were higher in the olanzapine-treated group than in the risperidone-treated group (OR=2.182, P=0.004), as were frequencies with two-fold (OR=3.064, P=0.017) and three-fold (OR=2.883, P=0.039) elevation. Recovery time was longer in the olanzapine-treated group than in the risperidone-treated group (P=0.0059), as was latency time (P=0.0044). These results suggest that there are potential differences in antipsychotic-associated hepatic enzyme alterations between risperidone and olanzapine treatment. Controlled, prospective studies should be conducted to identify the risk factors associated with an alteration in hepatic enzymes related to treatment with risperidone and olanzapine. PMID- 15812270 TI - The emergence of tics during escitalopram and sertraline treatment. AB - The selective serotonin reuptake inhibitors (SSRIs) have well-established antidepressant activity and have also demonstrated efficacy in a number of anxiety disorders, including panic disorder. The SSRIs, escitalopram and sertraline, are well-tolerated and movement disorders are rarely associated with their use. We present a case of the emergence of tics associated with escitalopram and sertraline treatment, which might be due to an imbalance in the dopaminergic neurotransmitter system. PMID- 15812271 TI - Gabapentin in the treatment of antipsychotic-induced akathisia in schizophrenia. AB - Antipsychotic-induced akathisia is characterized by subjective and objective motor restlessness, which is observed as a common extrapyramidal side-effect of antipsychotic agents. A patient is described who had antipsychotic-induced akathisia unresponsive to conventional therapy, and who began gabapentin therapy for insomnia. Significant improvement in his akathisia occurred when the gabapentin dose was increased, and his other treatment for akathisia was decreased and discontinued. Gabapentin may be effective by mechanisms similar to its action in restless legs syndrome and Parkinsonism, and/or via the GABA neurotransmitter system. PMID- 15812272 TI - Patient education workshop on CD-ROM: an innovative approach for staff education. AB - This article describes an innovative patient education workshop on CD-ROM for use by staff educators and patient education coordinators. The CD-ROM allows nurses to gain an in-depth understanding of patient education practice. With the interactive, computerized format, nurses can complete the workshop on their computers and at their convenience. The CD-ROM format uses streaming video clips, audio, thought questions, case studies, and interactive practice activities to enhance learning. PMID- 15812273 TI - Preceptor continuity in a nurse internship program: the nurse intern's perception. AB - The purpose of this study was to examine the new graduate's perception of preceptor continuity in regard to clinical performance, role transition, satisfaction, and retention in nursing. Determining how the new graduate perceives the role of a preceptor or multiple preceptors in a new graduate program is instrumental in evaluating and adapting the program to meet the needs of the new nurse graduate. PMID- 15812275 TI - A measure of success: nursing students and test anxiety. AB - Anxiety reactions are common to situations perceived as threatening; however, excessive anxiety may paralyze an individual and interfere with effective functioning. Nursing students and nursing orientees face multiple stressors as they progress through a competitive and oftentimes grueling nursing program or job orientation. This article will focus on the experience of test anxiety in nursing students and new orientees and measures the nurse educator may employ in reducing this anxiety to facilitate learner progression through a nursing program or orientation. PMID- 15812277 TI - Establishment of a hospital-based simulation skills laboratory. AB - Today's healthcare environment requires that nurses be prepared for increasingly complex patient populations. Simultaneously, managers and educators are challenged to provide competency verification programs and continuing education opportunities with fewer resources. Hospital clinical educators share a staff development initiative of launching a unique simulation skills laboratory. The laboratory is designed to ensure nurses can meet the needs of patients in today's healthcare arena. PMID- 15812278 TI - Recruiting licensed staff in mental health. AB - This 4- to 6-month mental health internship program for new graduates vocational nurses (GVNs) involves a 4-month mental health course, a new graduate course, management of assaultive behavior course, and a 1:1 preceptorship in mental health. This successful program has recruited and trained 18 VNs to improve licensed staff/patient ratios in mental health. These new graduates are licensed at the end of 4 months and are working successfully as licensed staff on the psychiatric units. All of these licensed vocational nurses (LVNs) are now completing prerequisites or entering registered nurse (RN) programs to earn an advanced degree. They bring their medical skills, positive attitudes, and therapeutic strategies to contribute to the milieu on the nursing units. They have reduced the facility's licensed staff shortage in mental health. PMID- 15812279 TI - Keeping SARS out: an education program for SARS screeners in one Ontario hospital. AB - On March 14, 2003, the Ontario (Canada) Ministry of Health and Long-Term Care alerted healthcare providers about four cases of atypical pneumonia resulting in two deaths within a single family in Toronto, Ontario. These cases were just the first of many that occurred in Toronto, Ontario, Canada, in the months to come. This article describes one community hospital's response to the severe acute respiratory syndrome (SARS) crisis and the important, although initially overlooked, role of staff development in keeping SARS out of this Ontario hospital. PMID- 15812280 TI - Limited math skills: a prescription for change. AB - New and recent graduates of schools of nursing have limited mathematical skills for medication administration. The purpose of this article is to provide a prescription for change to address the issue of limited math skills for medication administration. The authors recommend three areas of needed change: relationships, practice, and expectations. These three areas require attention from both the hiring agency and the educational institution. PMID- 15812282 TI - Treatment options for developmental dislocation of the hip after walking age. AB - Cases of developmental dislocation of the hip occur after walking age because of late or missed diagnosis and failed conservative or operative treatment. Although a general consensus exists for the treatment in infants up to the age of 18 months, the recommendations for the treatment of developmental dislocation of the hip in older children range from conservative treatment up to 3 years of age to primary surgical procedures from the age of 18 months. A review of the pertinent literature and of the different treatment options was performed and the authors' preferred treatment regime and techniques for the operative treatment of developmental dislocation of the hip after walking age are reported. PMID- 15812283 TI - Arthroscopic findings of the hip in new onset hip pain in adolescents with previous Legg-Calve-Perthes disease. AB - Nine adolescent patients with new onset hip pain and previous Legg-Calve-Perthes disease (LCPD) were evaluated by hip arthroscopy. Eight of the nine patients had previous hip surgery for their LCPD and a pain-free interval averaging over 5 years. Hip arthroscopy was performed in the supine position on a fracture table using a distention distraction technique. Eight of the nine hips had intra articular abnormalities including; four tears of the ligamentum teres, three femoral head osteochondritis dissecans, two labral tears, two hips with significant synovitis and three hips with other femoral head abnormalities. Seven of the eight hips improved following their arthroscopic treatment and the patients returned to their before-pain activities including sports. Three patients have required subsequent surgery. The abnormal mechanics from residual femoral head deformity may increase the risk of developing a variety of painful lesions. Hip arthroscopy is a safe way to evaluate and treat these intra articular abnormalities. PMID- 15812284 TI - Musculoskeletal conditions of acute leukemia and malignant lymphoma in children. AB - We retrospectively reviewed 16 patients who presented to hospitals with orthopaedic complaints. Twelve patients experienced initial symptoms in the extremities and four patients had back pain. The leukocyte count was elevated in one patient, decreased in two patients, and normal in 13 patients. On radiographic examination, osteopenia was observed in 10 patients, osteolytic lesions were observed in five, and pathologic fractures were observed in five. Because the initial presentation of patients with leukemia often involves the musculoskeletal system, orthopaedists need to recognize the symptoms of this disease to avoid misdiagnosis and to expedite the initiation of appropriate potentially lifesaving treatment. PMID- 15812285 TI - Acetabular augmentation induced by extracorporeal shock waves in rabbits. AB - We conducted this animal study to demonstrate whether exposing the acetabulum in immature rabbits to extracorporeal shock waves induces bone formation in the acetabulum. Five thousand shock waves of 100 MPa each were directed, from outside, at the acetabular roof of eight immature rabbits. At each of two time points (4 and 8 weeks) after treatment, the pelvises of four rabbits were removed and evaluated morphologically. Woven bone formation was observed on the lateral margin of the acetabular roof at 4 weeks after treatment, and the breadth of the acetabular roof in the coronal plane was significantly increased. Eight weeks after treatment, the woven bone disappeared; the breadth of the acetabular roof, however, was significantly increased. These findings demonstrated that extracorporeal shock waves induced acetabular augmentation in rabbits. We conclude that extracorporeal shock waves, perhaps, could be applied clinically for the treatment of acetabular dysplasia. PMID- 15812286 TI - Is prematurity important in ultrasonographic hip typing? AB - The aim of this study was to determine whether prematurity is important with respect to hip typing and planning of treatment/follow-up in newborns assessed with the Graf's ultrasonography method. Between January 1998 and 2003, 1592 neonates; preterm (n=432) and term (n=1160), had their hips ultrasonographically evaluated with Graf typing. There were no statistical differences between the two groups with respect to the numbers of hips that required follow-up assessment, or treatment. The results suggest that prematurity is irrelevant and use of chronological age as opposed to corrected age for prematurity will further simplify diagnosis and planning of follow-up and treatment in the Graf system for neonatal hip assessment. PMID- 15812287 TI - Valgus slipped capital femoral epiphysis. AB - Abnormally high stresses involving the capital femoral physis were found in a series of valgus slipped capital femoral epiphysis cases. Using a single leg stance model we studied 10 hips in seven patients in which the epiphysis was lateral and posterior in relation to the femoral neck and calculated that shear stresses were high enough to be associated with failure of the physis. Valgus neck shaft angles and lateral tilt of the physes were seen in all cases. PMID- 15812288 TI - Retrospective review of L3 myelomeningocele in three age groups: should posterolateral iliopsoas transfer still be indicated to stabilize the hip? AB - The data from 58 hips in 29 myelodysplastic children with L3 paralysis has been reviewed retrospectively. Posterolateral transfer of the iliopsoas (Sharrard technique) was performed on all hips to correct muscle imbalance between 1975 and 1992. The average length of follow-up was 21 years. Preoperatively, 58.6% of the hips were either subluxated or dislocated (these were surgically reduced at the same stage). Radiological assessment revealed that 91.4% of the hips were stable at preschool age and 84.5% in adolescence. Of the patients 86% were functional walkers at preschool age and 75.8% in adolescence. Downward transitions in seven patients were related to the presence of scoliosis, spasticity and obesity. All patients who were community walkers in adolescence maintained their function in adult life at the time of the review. We conclude that the procedure has value in obtaining hip stability and walking ability in carefully selected patients with myelomeningocele. PMID- 15812289 TI - Spinal cord sonography in newborns: anatomy and diseases. AB - Sonography of the spinal cord is a relatively recent acquisition that makes it possible to evaluate the content of the vertebral canal and study its pathologies. The aim of this study was to verify the reliability of ultrasound images by comparing them with magnetic resonance ones in healthy controls as well as in patients referred to us between 1991 and 2004. In this period, we studied 436 newborns: 88 without any suspicion of disease as normal controls, and 348 with suspected congenital diseases or in order to screen the children of diabetic mothers, a group that has shown an increased incidence of dysraphism. After explaining normal sonograms, we describe the pathological pictures observed in the 12 pathological cases in our series: conus hypomobility in five cases; lack of visualization of the conus medullaris in one case; and an enlarged ependymal canal in six cases. Four cases presented all three pathological conditions, and seven the association of two pathologies. All of these patients also underwent magnetic resonance imaging (MRI), which confirmed the ultrasound findings in four cases: three cases of enlarged ependymal canal and one of tethered cord hypomobility of the roots with an associated lipoma; the MRI findings were normal in the other seven cases. Sonography was highly specific but not very sensitive, because it is partially conditioned by patient collaboration. Nevertheless, subsequent MRI confirmed 37% of the suspected pathological cases. The ultrasound resolution of both normal and pathological spinal cord structures was particularly clear. The images were similar, easily comparable and often identical to the MRI results, although MRI was certainly more sensitive. The advantages of sonography are its non-invasiveness, low cost, the virtually ubiquitous availability of ultrasound equipment, the simplicity and rapidity of the examination, and its specificity. We believe that the indications for its use are lumbo-sacral skin alterations, neurological disorders caused by congenital malformations, traumas due to childbirth or a lumbar puncture, occult dysraphism, all of the compressive spinal cord neo-formations involving nerves, the dura mater and vertebral bone and joint structures, and the screening of the newborns of diabetic mothers. PMID- 15812290 TI - Morphology and dynamics of the ulnar nerve in the cubital tunnel after percutaneous cross-pinning of supracondylar fractures in children's elbows: an ultrasonographic study. AB - After the 12.6-month (3-30) follow-up period, the morphology and dynamics of the ulnar nerve in the cubital tunnel were examined with ultrasonography on operated (group 1) and opposite (group 2) elbows of 13 children who had percutaneous cross pin fixation for displaced supracondylar humerus fractures. When we compared group 1 and group 2, nerve morphology was similar (P>0.05), but nerve dynamics were found to be decreased or absent for seven (54%) elbows in group 1 (P<0.05). The medial K-wire may damage the ulnar nerve dynamics in the cubital tunnel. PMID- 15812291 TI - The role of the small AO external fixator in supracondylar rotational femoral osteotomies. AB - Torsional problems of the femur have been traditionally treated by a proximal osteotomy with internal fixation. We elected to perform femoral derotational osteotomies distally. Between September 1994 and April 2001, supracondylar osteotomies were performed on 38 femora in 21 children with torsional and angular deformities. The average age was 9 years (range 5-15 years). Twenty-three femora had excessive anteversion and fifteen, retroversion. All osteotomies were maintained by the small AO external fixator. Bony union occurred at an average of 10 weeks. Distal femoral osteotomy is an effective site for correcting rotational and associated angular deformities. The small AO external fixator provides precise adjustability, solid stability, and avoids a second procedure for hardware removal. PMID- 15812292 TI - The ski jump deformity of the distal phalanx: a new entity? AB - A case of a congenital hand anomaly is presented in which the distal phalanx of the little finger was curving dorsally in the sagittal plane. This may be the first reported case. A surgical approach to correct it is described. PMID- 15812293 TI - How closely do surgeons follow gait analysis recommendations and why? AB - Gait laboratory recommendations for surgery were compared with actual surgeries performed in 30 consecutive patients. The agreement between the procedures performed and those recommended by the gait laboratory averaged 93.2+/-13.4% overall and 86.0+/-18.3% excluding patients referred by the gait laboratory physician. For 23 patients (77%), the recommended surgeries exactly matched the surgeries ultimately performed. In the other patients, seven procedures that had not been recommended were performed, and seven procedures that had been recommended were not performed. Explanations for the differences included preoperative clinical or radiographic evaluation, intraoperative assessment, changes in function between gait analysis and surgery, and patient/family request. PMID- 15812294 TI - Gait characteristics of patients with bilateral club feet following posteromedial release procedure. AB - In this study we intended to evaluate the gait characteristics of children who had bilateral operated clubfeet, and to compare the results with gait parameters of children without clubfeet or lower extremity abnormalities. Fifteen children with bilateral plantigrade and asymptomatic clubfeet, operated bilaterally with posteromedial release method, were included in group P. Means+/-SD of age and follow-up time were 8.5+/-2.6 years (range, 4-14 years) and 7.7+/-2.8 years (range, 3-10 years), respectively. Age, sex, height-and weight-matched 24 children without clubfeet or lower extremity abnormalities were included in the control group (group C). Three-dimensional computerized gait analysis was performed for all subjects, and time-distance, kinematic and kinetic gait characteristics of the groups were compared. Time-distance characteristics of the groups were identical. Ankle plantar flexion angles, plantar flexor moment and generated power were significantly decreased in group P (P<0.05). Knee hyperextension during midstance was observed in 17 of 30 (57%) knees in the P group. Kinetic analysis revealed excessive knee valgus and flexion moments, as well as decreased second peak values of vertical ground reaction forces for group P. This study shows that even successfully operated, plantigrade and clinically asymptomatic children with bilateral clubfeet have significant gait deviations involving all lower limb joints. Future studies are needed to investigate the accountable factors of gait deviations and possible long-term musculoskeletal morbidity of operated children with bilateral clubfeet. PMID- 15812295 TI - Recurrence of aneurysmal bone cysts in young children: a multicentre study. AB - Some authors have reported that the clinical and pathologic behaviour of aneurysmal bone cysts (ABCs) is more aggressive in younger patients and that younger patients have more tumour recurrence. The authors carried out a retrospective, multicentred paediatric population-based analysis of 21 patients (14 boys and seven girls), 5 years of age or younger, with primary ABCs. Only patients with a minimum follow-up of 2 years were included. The most common operation was curettage (14 cases). Methylprednisolone acetate injection was used in two cases (failure in the initial diagnosis before biopsy) with negative results. An Ethibloc (Ethnor Laboratories/Ethicon, Norderstedt, Germany) injection was employed in four cases. There were five recurrences. Three lesions recurred once, one lesion recurred three times and one recurred six times. These recurrences occurred in two cases after methylprednisolone acetate injection, after Ethibloc (Ethnor Laboratories/Ethicon) injection (one case) and, after curettage (two cases). ABCs in children, 5 years of age or younger, do not seem to be more aggressive than in older children. Curettage is a surgical procedure that can be used even in young children. Of course, recurrence is always possible but the recurrence rate is not unacceptable. More aggressive operative intervention does not appear to be indicated. PMID- 15812296 TI - Quadricepsplasty in arthrogryposis (amyoplasia): long-term follow-up. AB - Eight patients with arthrogryposis multiplex congenita (amyoplasia type) (11 knees) with knee hyperextension deformity underwent quadricepsplasty and were analyzed during an average follow-up period of 11 years and 2 months. The results were clinically analyzed based on gait pattern, range of movement, and orthotic requirements. Joint congruency was evaluated by radiography according to the Leveuf Pais classification. A satisfactory result was the correction of the deformity, articular congruency, sufficient range of movement, adequate gait pattern and no need for orthosis. A satisfactory outcome occurred in five of the eight patients (eight knees). We considered an unsatisfactory result when any of these conditions occurred. Our experience demonstrated that the quadricepsplasty corrected the hyperextension deformity of the knee joint, improved function, gait pattern, and maintained the muscle power of the quadriceps. PMID- 15812297 TI - Rhabdomyosarcoma presenting as a calf pyomyositis. PMID- 15812298 TI - Good results after treatment with the Frejka pillow for hip dysplasia in newborn infants: a 3-year to 6-year follow-up study. PMID- 15812300 TI - Neuroinflammation in the pathogenesis of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis is a devastating motor neuron disorder. Traditionally regarded as a 'neuron only' disease, recent evidence suggested that other cells contribute critically to the pathogenesis. This review provides a short synopsis of the role neuroinflammation and microglial cells play in the disease and its animal models. A better understanding of neuroinflammation in motor neuron degeneration and amyotrophic lateral sclerosis disease progression promises to improve the rational design of greatly needed therapies. PMID- 15812301 TI - Mutant superoxide dismutase disrupts cytoplasmic dynein in motor neurons. AB - Cytoplasmic dynein and dynactin drive retrograde axonal transport in neurons, and mutations in dynein/dynactin cause motor neuron degeneration. To test whether defects in dynein/dynactin function are involved in the neurodegenerative disease amyotrophic lateral sclerosis, we examined neurotracer transport from muscle to motor neuron in a transgenic mouse model of amyotrophic lateral sclerosis. Significant inhibition was observed, which was temporally correlated with declines in muscle strength. No decrease in dynein/dynactin expression was observed, but immunohistochemistry suggests that dynein associates with aggregates of mutant Cu/Zn superoxide dismutase 1. Expression of mutant Cu/Zn superoxide dismutase 1 in primary motor neurons altered the cellular localization of dynein, suggesting an inhibition of dynein/dynactin function. Thus, inhibition of dynein/dynactin function may have a role in motor neuron degeneration in amyotrophic lateral sclerosis. PMID- 15812302 TI - Sex differences in pregnenolone sulphate in the chick brain after training. AB - Pregnenolone and pregnenolone sulphate are potent memory enhancers when administered to rodents prior to various learning and memory paradigms. Here, we show that training on a passive avoidance task results in the increased concentration of pregnenolone sulphate in the medial striatum, formerly known as lobus parolfactorius of female but not male chicks. In addition, we demonstrated potential for neuronal synthesis of pregnenolone in the day-old chick brain, including in the intermediate medial mesopallium, formerly known as intermediate medial hyperstriatum ventrale and the medial striatum. PMID- 15812303 TI - Structural brain changes in bipolar disorder using deformation field morphometry. AB - The objective of this study was to investigate anatomical brain abnormalities in adult bipolar patients using a deformation field morphometry technique. Our sample consisted of 32 right-handed bipolar I patients (men/women=16/16) and 32 right-handed, age and gender matched healthy controls. Deformation field morphometry analysis was performed on three-dimensional spoiled gradient recalled acquisition images. We found gender-specific structural differences between bipolar patients and healthy individuals. Bipolar men had significantly larger lateral ventricles (especially pronounced in the left hemisphere) and smaller left dorsolateral prefrontal cortex than healthy male controls. Our results are complementary to the findings of functional imaging and post-mortem studies that demonstrate abnormalities in the dorsolateral prefrontal cortex in bipolar patients. PMID- 15812304 TI - Averaged and single-trial brain responses in the assessment of human sound detection. AB - We investigated sound detection in humans with magnetoencephalography and behavioural measurements. Sounds with intensity increasing smoothly over 125-1000 ms elicited a transient response in auditory cortex with a peak latency in the 200-600 ms range. Importantly, peak latency accurately predicted behavioural reaction time and was unaffected by attentional engagement. Peak amplitude was augmented when the study participants attended to the stimuli and when stimulus duration was decreased. For investigating the cause of these amplitude variations in the averaged response we designed a wavelet-based method for analysing single trial responses. We found that attention affects the amplitude of the single trial responses whereas the intensity slope of the stimulus modifies their latency distribution. The transient response reported here holds promise for rapid, objective hearing assessment not requiring a behavioural task. PMID- 15812305 TI - Left-hemispheric brain activity reflects formant transitions in speech sounds. AB - Connected speech is characterized by formant transitions whereby formant frequencies change over time. Here, using magneto-encephalography, we investigated the cortical activity in 10 participants in response to constant formant vowels and diphthongs with formant transitions. All the stimuli elicited prominent auditory N100m responses, but the formant transitions resulted in latency modulations specific to the left hemisphere. Following the elicitation of the N100m, cortical activity shifted some 10 mm towards anterior brain areas. This late activity resembled the N400m, typically obtained with more complex utterances such as words and/or sentences. Thus, the present study demonstrates how magnetoencephalography can be used to investigate the spatiotemporal evolution in cortical activity related to the various stages of the processing of speech. PMID- 15812306 TI - Perceptual categorization of sound spectral envelopes reflected in auditory evoked N1m. AB - Magnetic responses to periodic complex sounds with equivalent acoustic parameters except for two different fundamental frequencies (F0) and 12 different spectral envelopes of vocal, instrumental, and linear shapes were recorded to determine the cortical representation of timbre categorization in humans. Responses at approximately 100 ms (N1m) to vocal and instrumental (nonlinear) sounds were localized significantly anterior to linear sound responses. N1m source strength for nonlinear sounds was significantly larger than that for linear sounds, and this difference was more marked in the left hemisphere than in the right. N1m peak latency only for vocal sounds was not affected by F0. Perceptual categorization was reflected in N1m source strength and location (linear or nonlinear), and in N1m latency (vocal or nonvocal). PMID- 15812307 TI - Electrophysiological evidence for lateralization of preparatory motor processes. AB - Event-related potentials were recorded during a warned choice reaction time task in which the warning signal was informative (it signaled the specific response to be required) or uninformative (it gave no response-specific information). The response signal specified a key press with either the middle or the ring finger of the left or the right hand. The informative warning signal facilitated reaction time in two separate protocols. The contingent negative variation elicited following the informative warning signal was larger than that following the uninformative warning signal, and this effect was larger over the left hemisphere than over the right hemisphere. This finding gives converging evidence that preparatory motor processes are predominantly lateralized to the dominant hemisphere. PMID- 15812308 TI - Medullary N-type and P/Q-type calcium channels contribute to neuropathy-induced allodynia. AB - The present study was designed to determine the contribution of N-type, P/Q-type and L-type calcium channels in the rostral ventromedial medulla to tactile allodynia following peripheral nerve injury. L5/L6 spinal nerve ligation in rats produced tactile allodynia, which was dose-dependently inhibited by intrarostral ventromedial medulla microinjection of the N-type calcium channel antagonist omega-conotoxin MVIIA. Similarly, intrarostral ventromedial medulla microinjection of the P/Q-type calcium channel antagonist omega-agatoxin IVA inhibited spinal nerve ligation-induced tactile allodynia, whereas intrarostral ventromedial medulla microinjection of the L-type calcium channel antagonist nimodipine had no effect. These results demonstrate that N-type and P/Q-type calcium channels in the rostral ventromedial medulla contribute to tactile allodynia following peripheral neuropathy, likely via neurotransmitter-mediated activation of descending facilitatory systems from the rostral ventromedial medulla. PMID- 15812309 TI - Neural mechanisms of attentional modulation of perceptual grouping by collinearity. AB - Psychophysical research showed that detection of an oriented visual target is facilitated when the target is grouped with collinear visual flankers. However, this collinear grouping effect is evident only when the flankers are attended. This study examined neural mechanisms underlying the interaction between attention and grouping by collinearity. Event-related potentials were recorded from study participants who judged whether oriented Gabor patches (i.e. visual elements consisting of a sinusoidal contrast modulation convolved with a Gaussian function) along the cued orientation were collinear or orthogonal. Event-related potentials showed an enhanced negativity over the posterior occipital cortex at 48-72 ms when collinear patches were congruent rather than incongruent with the cued orientation. A negative shift between 260 and 380 ms was observed over the occipital-parietal areas in the congruent rather than incongruent conditions. The long-latency effect, however, was evident only when the collinear patches were allocated along 45 degrees . The event-related potential results suggest that the interaction between attention and collinear grouping may take place as early as in the primary visual cortex and is independent of global orientations of perceptual groups. PMID- 15812310 TI - Analysis of cerebellum proteomics in the hydrocephalic H-Tx rat. AB - The H-Tx rat is a polygenic inherited model of hydrocephalus. In order to identify disease-specific biomarkers associated with congenital hydrocephalus, comparative proteomic analysis was used to screen cerebellum proteins in H-Tx rats at 1 day after birth. Seven proteins showed significant changes in hydrocephalic H-Tx rats compared with Sprague-Dawley and normal H-Tx rats, including HMG-1, CDCrel-1A, mitochondrial ATP synthase, ERp29, NADP+-ICDH, CCT beta and gamma. This indicates that the hydro-cephalus in H-Tx rats may be the result of a panel of proteins. In particular, the presence of HMG-1 and lack of CDCrel-1 in hydrocephalic H-Tx rats suggests that the use of two protein markers will provide further insight with respect to congenital hydrocephalus. PMID- 15812311 TI - Characterization of a cyclooxygenase-2-765G-->C promoter polymorphism in human neural cells. AB - Direct sequencing of the human cyclooxygenase-2 gene promoter revealed a common single nucleotide substitution, cyclooxygenase-2-765G-->C, in 24.5% of the populations analyzed. This change introduced a 20 base pair polypyrimidine/polypurine element and a partial recognition feature for RXRalpha, the 9-cis retinoic acid receptor, into the polymorphic promoter. Cyclooxygenase-2 765G-->C constructs, when transfected into human neural cells, exhibited a 1.4 fold higher level of basal expression, while the proinflammatory factors interleukin-1beta and 9-cis retinoic acid synergistically induced polymorphic promoter activity 2.4-fold over wild type. These results suggest that under specific conditions of cellular stress, a common variation in cyclooxygenase-2 promoter structure may enhance cyclooxygenase-2 transcription, and this may contribute to the proliferation of an inflammatory response in brain cells. PMID- 15812312 TI - Glial cell line-derived neurotrophic factor in bone marrow stromal cells of rat. AB - To investigate the capability of Sprague-Dawley rat bone marrow stromal cells to secrete glial cell line-derived neurotrophic factors (GDNF), we detected expression of GDNF messenger RNA and protein in bone marrow stromal cells of Sprague-Dawley rats by reverse-transcriptase polymerase chain reaction and enzyme linked immunosorbent assay (ELISA), respectively. The GDNF messenger RNA and protein were detected in culture medium and total cell protein when bone marrow stromal cells were cultured for 3 days. The levels of GDNF in culture medium and total cell protein increased gradually after 3, 7 and 10 days of culture. Rat bone marrow stromal cells have the potential to secrete GDNF. Furthermore, the ability of secretion is determined by the surrounding microenvironment and self growth status. PMID- 15812313 TI - Pyruvate protects motor neurons expressing mutant superoxide dismutase 1 against copper toxicity. AB - Mutations in the copper/zinc superoxide dismutase (SOD1) gene are known to be responsible for familial amyotrophic lateral sclerosis. Alteration of metal binding properties of mutant SOD1 has been proposed to play a role in the pathogenesis of amyotrophic lateral sclerosis. We investigated the toxic effects of excess extracellular copper on motor neuronal cells expressing human mutant SOD1 (G93A), and evaluated the neuroprotective effects of energy metabolism intermediates or cofactors. Motoneuron-neuroblastoma hybrid (VSC 4.1) cells expressing mutant SOD1, when treated with copper chloride, showed reduced viability and increased levels of endogenous peroxides. Moreover, this copper induced toxicity was attenuated by a free radical scavenger, a caspase inhibitor, or a calpain inhibitor. Of the energy metabolism intermediates examined, pyruvate significantly reduced the death and production of reactive oxygen species in cells expressing mutant SOD1. Our data suggest that pyruvate could be of therapeutic value in some forms of familial amyotrophic lateral sclerosis. PMID- 15812314 TI - Zinc induces neurofilament phosphorylation independent of p70 S6 kinase in N2a cells. AB - Hyperphosphorylated neurofilaments are a part of neurofibrillary tangles in Alzheimer's disease brains. Zinc has been shown to be increased in the brain areas heavily affected by Alzheimer pathologies. Zinc could induce tau hyperphosphorylation in SH-SY5Y and N2a cells, and tau phosphorylation may be mediated by p70 S6 kinase activation. Many of the tau kinases can also phosphorylate neurofilaments, and in this study we wanted to see whether neurofilament phosphorylation is regulated by p70 S6 kinase in N2a cells. We found that zinc induces rapamycin-dependent p70 S6 kinase phosphorylation at Thr421/Ser424 and Thr389, and rapamycin-independent phosphorylation of neurofilaments at the SMI34 epitope. Although zinc could induce cell proliferation and cell growth, and increased phosphorylation of neurofilaments, only cell growth appeared to be related to p7056kinase activation. PMID- 15812315 TI - Inhibition of glycogen synthase kinase-3 protects cells from intrinsic but not extrinsic oxidative stress. AB - Oxidative stress is linked to neuronal dysfunction and death in many diseases. Glycogen synthase kinase-3 often promotes apoptosis, so this investigation tested whether glycogen synthase kinase-3 is linked to oxidative stress-induced apoptosis. Both intrinsic oxidative stress induced by the mitochondrial inhibitor rotenone and extrinsic oxidative stress induced by exogenously added H2O2 activated Bax, caspase-2, and caspase-3 in human neuroblastoma SH-SY5Y cells. Inhibitors of glycogen synthase kinase-3 blocked rotenone-induced, but not H2O2 induced, activation of both caspases, but not Bax activation. Thus, glycogen synthase kinase-3 is an important component of intrinsic oxidative stress-induced apoptosis that acts downstream of mitochondrial Bax activation, and there are substantial differences in the role of glycogen synthase kinase-3, and lithium's effects, in apoptotic signaling induced by intrinsic and extrinsic oxidative stress. PMID- 15812316 TI - Age-related effects of bromocriptine on sensory gating in rhesus monkeys. AB - Declines in dopamine neurotransmission are a robust characteristic of the process of normal aging. Using neuroimaging, biochemical and cognitive methods, age related reduction of D2 receptor has been noted in a wide range of species. On the other hand, it is well known that dopamine plays a crucial role in the modulation of sensory gating. Here, we examined age-related alterations of D2 receptor in rhesus monkeys, using a sensory gating paradigm. The direct D2 receptor agonist, bromocriptine, was characterized in young adult and aged monkeys. We found bromocriptine disrupted sensory gating in young adult monkeys but not in aged ones. Our results provided new evidence that there is a functional decline of D2 receptor in aged monkeys. PMID- 15812317 TI - Brain correlates of sentence translation in Finnish-Norwegian bilinguals. AB - We measured brain activation with functional magnetic resonance imaging (fMRI) while Finnish-Norwegian bilinguals silently translated sentences from Finnish into Norwegian and decided whether a later presented probe sentence was a correct translation of the original sentence. The control task included silent sentence reading and probe sentence decision within a single language, Finnish. The translation minus control task contrast activated the left inferior frontal gyrus (Brodmann's area 47) and the left basal ganglia. The left inferior frontal activation appears to be related to active semantic retrieval and the basal ganglia activation to a general action control function that works by suppressing competing responses. PMID- 15812318 TI - Sexually dimorphic link between dopamine D2 receptor gene and neuroticism anxiety. AB - Prior theory-driven research probing the association between dopaminergic candidate genes and human personality has focused on the trait of novelty seeking. Here, we examined the association between the dopamine D2 receptor (DRD2) TaqI A polymorphism and two other personality traits, neuroticism-anxiety and agentic extraversion. We found no significant associations for agentic extraversion. However, for men, but not for women, we observed a strong and specific association between low neuroticism-anxiety and the A1+ allele of the DRD2 TaqI A polymorphism across two independent samples and across two alternative personality scales. We conclude that new theoretical models are needed to account for these and other recent reports of associations between neuroticism-anxiety and brain dopamine, which cannot be interpreted within the traditional framework. PMID- 15812319 TI - Left prefrontal repetitive transcranial magnetic stimulation impairs performance in affective go/no-go task. AB - Functional neuroimaging studies have associated affective go/no-go function with lateral prefrontal activation, but they have not established a causal role and have not determined whether one hemisphere is predominantly engaged. In the present study, 11 normal volunteers underwent slow repetitive transcranial magnetic stimulation of the left and right dorsolateral prefrontal cortex, and the occipital cortex prior to performance of a picture-based affective go/no-go task. We found an interfering effect of left prefrontal repetitive transcranial magnetic stimulation compared with both right prefrontal and occipital repetitive transcranial magnetic stimulation. This impairment concerned positive and negative task stimuli to a similar extent, and tended to be greater in shift compared with nonshift blocks. Our findings demonstrate a functionally relevant lateralization of the prefrontal contribution to affective go/no-go tasks. PMID- 15812320 TI - A functional magnetic resonance imaging study of the tradeoff between semantics and phonology in reading aloud. AB - Using functional magnetic resonance imaging, we explored the role of semantics in mediating orthographic-to-phonological processing in reading aloud, focusing on the interaction of imageability with spelling-to-sound consistency for low frequency words. Behaviorally, high-imageable words attenuate the standard latency and accuracy disadvantage for low-frequency inconsistent words relative to their consistent counterparts. Neurobiologically, high-imageable words reduced consistency-related activation in the inferior frontal gyrus but increased posterior activation in the angular and middle temporal gyri, representing a possible neural signature of the tradeoff between semantics and phonology in reading aloud. We discuss implications for neurobiological models of reading in terms of understanding the interplay among areas associated with component processes and suggest that the results constitute an important step toward integrating neurobiological and computational models of reading. PMID- 15812321 TI - Relationship between steady-state and induced gamma activity to motion. AB - When a moving stimulus is presented at a specific temporal frequency, both steady state responses and induced gamma activity may be elicited in the electroencephalogram. The electroencephalogram was recorded when study participants viewed random dot kinematograms under three conditions: coherent motion, incoherent motion and stationary. Dot position was changed at a rate of 9.3 Hz in the coherent and incoherent conditions. Induced power at 40 Hz was increased during coherent motion compared with the other conditions. In contrast, the steady-state response at 9.3 Hz showed a trend for increased power during the incoherent condition. These results suggest that steady-state responses to moving stimuli reflect sensory activation, while the induced gamma activity indexes perceptual processes. PMID- 15812322 TI - Global and local music perception in children with Williams syndrome. AB - Musical processing can be decomposed into the appreciation of global and local elements. This global/local dissociation was investigated with the processing of contour-violated and interval-violated melodies. Performance of a group of 16 children with Williams syndrome and a group of 16 control children were compared in a same-different task. Control participants were more accurate in detecting differences in the contour-violated than in the interval-violated condition while Williams syndrome individuals performed equally well in both conditions. This finding suggests that global precedence may occur at an early perceptual stage in normally developing children. In contrast, no such global precedence is observed in the Williams syndrome population. These data are discussed in the context of atypical cognitive profiles of individuals with Williams syndrome. PMID- 15812323 TI - Sex differences in the preattentive processing of vocal emotional expressions. AB - Emotional tone of voice indicates the significance of a spoken utterance. We asked whether listeners recognize this significance even when attending to something else and whether men and women differ in this regard. To answer these questions, we presented emotionally or neutrally spoken syllables as standards and deviants in a mismatch negativity paradigm. Independent of the listeners' sex, deviants elicited a mismatch negativity in the scalp-recorded event-related potential as an indicator of preattentive acoustic change detection. Only women, however, showed a larger mismatch negativity to emotional than to neutral deviants. Thus, even though both sexes detect change in voice preattentively, only women recruit additional processing resources when the change in voice is one of emotional valence. PMID- 15812324 TI - An event-related potential investigation of the time-course of temporal ventriloquism. AB - Temporal ventriloquism refers to the phenomenon that a sound presented in close temporal proximity of a visual stimulus attracts its perceived temporal occurrence. Here, we investigate the time-course of the neuronal processes underlying temporal ventriloquism, using event-related brain potentials. To measure shifts in perceived temporal visual occurrence, we used a paradigm in which a sound modulates the magnitude of a visual illusion called the flash-lag effect. A sound presented before the flash reduced both the size of the flash-lag effect and the amplitude of visual N1 compared with when the sound lagged the flash. We attribute the modulation of the flash-lag effect to a modulation of facilitation of visual processing. The time-course (190 ms) and localization (occipitoparietal cortex) of this particular auditory-visual interaction confirms the sensory nature of temporal ventriloquism. PMID- 15812325 TI - Sentence processing in 30-month-old children: an event-related potential study. AB - In a previous event-related brain potential study, we provided evidence that preschoolers display different brain electrical patterns to semantic content and syntactic structure processing. In the present study, we aimed to determine the time-course of these event-related potential effects in 30-month-old children, using the same syntactically anomalous, semantically anomalous and control sentences that we used in our previous study. The results show that semantic violations elicit a frontal negativity peaking around 600 ms, whereas the morphosyntactic violations elicit a slow positive shift peaking around 800 ms with a frontocentral distribution. Our findings replicate the event-related potential patterns previously observed in young children and indicate that the neural signatures of sentence processing can be observed at an early point in development. PMID- 15812326 TI - Motion verb sentences activate left posterior middle temporal cortex despite static context. AB - The left posterior middle temporal region, anterior to V5/MT, has been shown to be responsive both to images with implied motion, to simulated motion, and to motion verbs. In this study, we investigated whether sentence context alters the response of the left posterior middle temporal region. 'Fictive motion' sentences are sentences in which an inanimate subject noun, semantically incapable of self movement, is coupled with a motion verb, yielding an apparent semantic contradiction (e.g. 'The path comes into the garden.'). However, this context yields no less activation in the left posterior middle temporal region than sentences in which the motion can be applied to the subject noun. We speculate that the left posterior middle temporal region activity in fictive motion sentences reflects the fact that the hearer applies motion to the depicted scenario by scanning it egocentrically. PMID- 15812327 TI - Lexical priming and semantic integration reflected in the event-related potential of 14-month-olds. AB - This study investigates by means of the event-related brain potential whether mechanisms of lexical priming and semantic integration are already developed in 14-month-olds. While looking at coloured pictures of known objects children were presented with basic-level words that were either congruous or incongruous to the pictures. The event-related potential of 14-month-olds revealed an early negativity in the lateral frontal brain region for congruous words, and a later N400-like negativity for incongruous words. These results indicate that both lexical priming and semantic integration are already present as early as 14 months. PMID- 15812328 TI - Carve out time to think--yes, think. PMID- 15812332 TI - Coming soon: Tissue storage and issuance standards. AB - Nurse managers will be responsible for ensuring compliance with JCAHO's newest set of standards. PMID- 15812333 TI - Better retention through nursing theory. PMID- 15812334 TI - The new manager's guide to surviving (and thriving). AB - Consider the top 10 ways new managers can successfully fulfill their roles. PMID- 15812336 TI - 101 tips to retain a happy, healthy staff. AB - Nurse managers share easy-to-implement ways to improve retention in the unit. PMID- 15812338 TI - Build a framework for conflict assessment. AB - Review a framework for assessing organizational conflict in a case study format that illustrates implementation strategies. PMID- 15812339 TI - Helping hands: CNAs in elder care. AB - One facility's functional model of care for hospitalized elders focuses on improving nutrition, toileting, and promoting mobility. PMID- 15812340 TI - The Magnetic pull. AB - Two hospitals share their specific Forces of Magnetism. PMID- 15812341 TI - Patient satisfaction: get the EDge. AB - Due to increasing competitiveness in healthcare, emergency department personnel should focus on patient satisfaction initiatives. PMID- 15812342 TI - Does your staffing agency have JCAHO's stamp of approval? AB - Travel companies react to JCAHO's new Health Care Staffing Services Certification. PMID- 15812343 TI - Is electromagnetic interference still a risk? AB - Signs banning all cell phone use are still ubiquitous in hospitals, but recent studies show that global bans may not be necessary. PMID- 15812346 TI - Health fairs extend your outreach. PMID- 15812349 TI - Integrin receptors and extracellular matrix components in parotid neoplasms. AB - In the review, the role of integrin receptors as well as of some of their ligands of the extracellular matrix in cases of parotid neoplasms is discussed, in particular, the data concerning the expression of various types of integrin adhesion molecules in benign and malignant tumors of the parotid, the expression of fibronectin, tenascin, and collagen type IV etc. The role of dysregulation of the expression of these proteins in invasion and metastasis of parotid tumors is discussed. PMID- 15812350 TI - Up-date on solitary plasmacytoma and its main differences with multiple myeloma. AB - Solitary plasmacytoma is plasma cell neoplasm. It is a localized bone disease and for this reason it is different from multiple myeloma (systemic plasma cell neoplasm). Sometimes, solitary plasmacytoma precedes a following multiple myeloma. Clinical findings of solitary plasmacytoma are related to the univocal localization on damaged bone, while laboratory findings could be similar to multiple myeloma (i.e. M component, kidney dysfunction, blood calcium alterations, increased beta-2-microglobulin). However, during a solitary plasmacytoma, laboratory findings could not be present contemporaneously such clinical complications (i.e. kidney failure, immunological disorders with a trend toward infectious disease and/or autoimmunity, neurological disorders, haematological disorders, amyloidosis, POEMS syndrome). These raise the reason because solitary plasmacytoma has better prognosis compared to multiple myeloma. PMID- 15812351 TI - Quality of life survey in patients with advanced prostate cancer. AB - Prostate cancer is a major health concern for Western patients who are facing important decisions, alongside with the physicians, concerning their treatment. Additionally, the physician has to take into account the potential side effects of the disease and the treatment itself. Treatment for advanced prostate cancer produces problematic physical and psychosocial side effects. The patient should be asked by the physician for information concerning daily life activities, overall satisfaction voiding ability, and sexual activity in a standardized questionnaire assessing the quality of life (QoL) of the patient. Many such questionnaires and studies have been evaluated or are still in progress. Modern QoL questionnaires are considered to be more than a necessity nowadays for the improvement of the treatment administered from the physician and the performance and functional status of the patient. PMID- 15812352 TI - Selection and characterization of heat-resistant variants of multidrug-resistant human gastric carcinoma cell lines. AB - AIM: To generate heat resistant variants selected from established human gastric carcinoma cell lines exhibiting different types of multidrug resistance (MDR) phenotype, i.e. EPG85-257P, the drug sensitive parental cell-line, EPG85-257RDB, a classical MDR subline and EPG85-257RNOV, which is an atypical multidrug resistant subline. METHODS: Thermoresistance was induced by stepwise increase of the growth temperature from 37.0 to 39.4 degrees C. Thermoresistance was determined by change of population doubling time (PDT) and clonogenic survival after acute hyperthermia at 42, 43, 44 and 45 degrees C. RESULTS: Most of the cells exhibited necrosis at elevated culture temperature. The PDT of the surviving thermoresistant variants were increased two-fold (EPG85-257P-TR) and 1.2-fold (EPG85-257RNOV-TR), respectively. No PDT change was observed with the lowest thermoresistant subline EPG85-257RDB-TR. Dose response curves after acute hyperthermia indicated a stable increase of thermotolerance of the parental cell line and the atypical MDR subline (50-90-fold at 45 degrees C), but not of the classical MDR subline, which was only increased at 43 degrees C (3-4-fold). Acquired thermoresistance did not change after freezing/thawing procedures. CONCLUSION: All cell lines achieved chronically induced thermoresistance. Thermotolerance after acute hyperthermia was present in the drug sensitive parental cell line and the atypical MDR subline, but not in cells exhibiting a classical MDR phenotype. PMID- 15812353 TI - The role of CD150-SH2D1A association in CD150 signaling in Hodgkin's lymphoma cell lines. AB - AIM: To find out what signal transduction pathways are linked to CD150 in Hodgkin's lymphoma (HL) cell lines, how are they regulated, and to examine the expression of different CD150 splice forms and SH2 domain containing protein D1A (SH2D1A) adaptor protein on mRNA level in HD primary tumors and cell lines. METHODS: The expression of CD150 splicing forms and SH2D1A adaptor protein in HD primary lymphoma tissue and cell lines were analyzed by RT-PCR method. To examine CD150-SH2D1A localization in HD cell lines we performed double immunofluorescent staining of these two proteins. Total amount of SH2D1A, Syk, Lyn, SHP-2, SHIP proteins, and activated/phosphorylated ERK1/2 and Akt proteins were detected by Western blot analysis with specific antibodies. RESULTS: We showed the expression of soluble (sCD150) and full length transmembrane (mCD150) splice forms and SH2D1A adaptor protein on mRNA level in 9 cases of classical HD and three HD lines of B cell origin - L428, KM-H2 and L1236. In spite of CD150 and SH2D1A co expression in studied HD cell lines, CD150 co-precipitated and co-localized with SH2D1A only in L1236 cells. CD150 ligation induced extracellular signal-regulated kinases (ERK) dephosphorylation in L1236 cell line, but had no effect on ERK pathway in KM-H2 and L428 cells. CD150 crosslinking induced Akt activation also only in L1236 cells. CONCLUSIONS: HD cells express sCD150 and mCD150 splice forms and SH2D1A. Association of CD150 and SH2D1A depends at least on their localization pattern. CD150 is linked to ERK and Akt pathways in HD cell lines. Our data suggest that CD150-SH2D1A association play decisive role in Akt signaling upon CD150 ligation in HD cell lines. CD150-mediated Akt activation in HD cell lines, similarly to DT40 model system, is SHIP-independent. PMID- 15812354 TI - Transient activation of Ras-dependent signalling at the early stages of Herbimycin A induced erythroid differentiation of human K562 cells. AB - AIM: To study the dynamics of Ras-dependent signalling in the course of Herbimycin A induced erythroid differentiation of human erythroleukemia K562 cells. METHODS: p21Ras functional activity was analyzed by direct measurement of GTP/GDP ratio in anti-p21Ras immunoprecipitates of K562 cells previously incubated with H3(32)PO4. Dynamics of protein tyrosine phosphorylation was studied using Western blotting. Electrophoretic mobility shift assay was used to monitor Erk2 activation. Phosphotyrosine (pY)-containing proteins bound to recombinant glutathione-S-tranferase (GST)-fused form of adaptor protein Grb2 were identified using GST in vitro binding assay. RESULTS: It was shown that the relative quantity of GTP associated with Ras protein in non-induced cells varied from 27% to 37% upon 72 h of cell culturing. Herbimycin A caused 15% increase of GTP/GDP ratio at 3rd h. This index decreased during further investigated periods, although it did not reach control values even at 72nd h. Transient rise of Ras GTP level at 3rd h of incubation in the presence of Herbimycin A correlated with the increase in tyrosine phosphorylation of proteins with apparent molecular weight of 210, 160, 140, 116 and 42 kDa, as well as with the activation of Erk2 and increase of binding of a set of pY-containing proteins with recombinant GST fusion form of Ras activator, adaptor protein Grb2. Dramatic inhibition of interaction between docking protein Shc and GST-Grb2 was observed at late stages of cell induction (48-72 h) while binding of pY-containing proteins during this period did not differ significantly in control and differentiated cells. CONCLUSION: The obtained results suggest that time-dependent changes in Grb2 mediated network of protein-protein interaction events might define implication of Ras-dependent signalling in Herbimycin A-induced erythroid differentiation of K562 cells. PMID- 15812355 TI - Fragility of telomeres after bleomycin and cisplatin combined treatment measured in human leukocytes with the Comet-FISH technique. AB - THE AIM of the present study was the comparative investigation of action of widely applied anticancer preparations: cisplatin (cis-DDP) and bleomycin (BLM) on total DNA and telomeres damage in human blood cells. METHODS: The "Comet-FISH technique" -- single cell gel electrophoresis ("comet assay") in combination with fluorescent in situ hybridization (FISH) was used for this purpose. This newly applied combined approach permits to detect on the same specimen the total DNA damage in individual cells and evaluate specific DNA sequences as well. Telomere - specific -- PNA (peptide nucleic acid) probes were used for the localization of telomeres in the comet's head and their migration to the tail. THE RESULTS obtained indicate that in control variants, due to DNA metabolism and handling, approximately 7% of the DNA and 17% of the telomeres were found in the tail. In cells treated with BLM alone, telomeres leak out with equal probability as total DNA. In turn, the combination of cis-DDP with BLM reduces telomere migration more than the migration of total DNA due to cis-DDP crosslinking effect. Thus, preferentially telomeric action of the cis-DDP can be concluded. CONCLUSION: The Comet-FISH approach permitted us to reveal the induction of DNA breaks with BLM and its modification due to platinum-crosslink formation, using telomeric PNA probes. PMID- 15812357 TI - Accumulation of chlorine e6 derivatives in cells with different level of expression and function activity of multidrug resistance protein P-gp 170. AB - AIM: This work deals with studying the influence of overexpression and function activity of multidrug transporter protein P-gp 170 on the intracellular accumulation of several porphyrin sensitizers, including chlorin e6 (Chl e6), di- (DME) and trimethyl esters (TME) of Chl e6. METHODS: A parental IM9 cell line and two IM9 drug-resistant IM9 sublines were used. With flow cytometry technique the rates of chlorines accumulation in different IM9 cells and values were related to the extent of multidrug resistance (MDR) phenotype. RESULTS: Using P-gp 170 specific antibodies and fluorescent probe JC-1 an increased expression and function activity of P-gp 170 was detected for drug-resistant cells. It was obtained that drug-resistant IM9 cells accumulated chlorines to a lesser extent than the respective wild type, however the differences did not exceed 30%. Verapamil, cyclosporine, known to reverse the MDR phenotype affects equally IM9 Vinc IM9-Tax and Im9 cells. CONCLUSION: Our results demonstrate that the P-gp 170 does not appear to play a role in the intracellular accumulation of chlorines. Since the enhanced activity of P-gp 170 in tumor cells is a factor of their resistance to the action of various antitumor drugs, we conclude that photodynamic therapy could be useful in killing cells exhibiting the MDR. PMID- 15812356 TI - Attempts to influence the drug resistance of tumor cells in experimental system. AB - AIM: Analysis of two approaches to the problem of multidrug resistance management in the course of tumor treatment by working out an experimental model relevant to clinical research. METHODS: Four cell sublines resistant to widely used cytostatics taxotere (Tax) or vincristine (Vcr) were developed by long-term culturing of the parental lymphoblastoid cell line IM-9 with incremental doses of these drugs. Constant presence of cyclosporin A (CsA) as a P-glycoprotein (Pgp) blocker in some cell cultures simulated prophylactic inhibition of this protein activity for prevention of drug resistance development from the very beginning of treatment. Both types of resistant cell sublines (developed in the presence or absence of cyclosporin A) were undergone to the action of the same Pgp blocker and drug inducing agents in short time cultures. Pgp activity by flow cytometry with fluorescence dye retention was checked and cytotoxicity assay by MTT-test was performed for drug resistance. RESULTS: According to the experimental data obtained drug resistance prophylactics policy is problematically effective in prevention of tumor cells resistance to cytostatics. However, the attempts to influence drug resistance by Pgp pharmacologic blockade since it developed are much less benefit in case of the preceding preventive treatment. PMID- 15812358 TI - 2-[(3-carboxy-1-oxoprogy1) amino]-2-deoxy-D-glucose induces apoptosis in Hep G2 cells. AB - AIM: To determine the effect of 2-[(3-carboxy-1-oxoprogy1) amino]-2-deoxy-D glucose (COPADG) on proliferation and apoptosis of human hepatocellular carcinoma cells (Hep G2). METHODS: Hep G2 cells were cultured in RPMI-1640 medium in the presence of various concentrations of COPADG. Cell proliferation was determined by MTT assay. Apoptosis was determined by fluorescence microscopy, transmission electron microscopy, agarose gel electrophoresis of DNA fragmentation, and flow cytometry. RESULTS: At the concentration ranging between 1-30 microM, COPADG potently inhibits growth and induce apoptosis of Hep G2 cells. CONCLUSIONS: COPADG could effectively induce apoptosis in Hep G2 cells. It may be potentially useful as a new agent for treatment of human hepatocellular carcinoma. PMID- 15812359 TI - A novel approach to tumor suppression using microencapsulated engineered J558/TNF alpha cells. AB - AIM: Immunoisolation technology using microencapsulated nonautologous cells is a novel alternative approach to the treatment of cancer. The work was aimed on investigation of the effect of implantation of microencapsulates on tumor growth in vivo. METHODS: In this study, we constructed an engineered tumor cell line J558/TNF-alpha that secreted functional tumor necrosis factor-alpha (TNF-alpha) (2 ng/ml), and went on to encapsulate the engineered cells into microencapsules. RESULTS: Our data showed that the microencapsulates thus produced could release functional TNF-alpha (1.2 ng/ml), which then diffused through the microencapsule membrane into the supernatant, and produced a cytotoxic effect on L929 cells in vitro. Microencapsulated cells were intratumorally (i.t.) implanted into athymic nude mice bearing the human breast cancer MCF-7. The results showed that the i.t. implantation induced extensive tumor cell apoptosis and necrosis leading to significant tumor regression and slower tumor growth than in the control groups that were i.t. injected with microencapsulated J558 or PBS respectively (p < 0.05). CONCLUSION: This study provides further evidence that the microencapsulation of recombinant tumor cells secreting cytokines may be an alternative approach in treatment of cancer. PMID- 15812360 TI - Whole body hyperthermia supplemented with urotropin in the treatment of malignant tumors. AB - AIM: To evaluate the effect of urotropin on the tolerance of an extreme whole body hyperthermia (WBH) regime (44.5-45.0 degrees C for 60 min) in rats, and then its application to pediatric patients with advanced cancer during WBH sessions (42.5-43.0 degrees C). METHODS: In experiments nonbred rats bearing Sarcoma 45 (Sa 45) were subjected to severe WBH with and without urotropin. Tolerance of WBH as well as tumor growth and survival of animals were monitored. Extreme WBH sessions (42.5-43.0 degrees C) + urotropin were used in multimodal treatment of 13 children with advanced, refractory or recurrent malignant tumors (42 procedures in total). RESULTS: Our experiments showed 5-fold increase in rat survival during the first two days following application of extreme WBH due to the additional administration of urotropin. This regime demonstrated a significant inhibitory effect upon the growth of Sa 45. In children with progressive malignant tumors we achieved extreme WBH regimes (up to 43.2 degrees C) using urotropin without additional toxicity and with 69% overall response rate. CONCLUSION: Extreme WBH (42.5-43.0 degrees C) could be used in the management of advanced and refractory cancer. Urotropin may play a favorable role during the procedures of extreme WBH, decreasing thermal damage to the body. Lymphocyte collection by lymphopheresis before WBH session and its reinfusion may be one of the progressive approaches for minimizing lymphocyte apoptosis. PMID- 15812361 TI - Effect of environmental nitric oxides on the antitumor resistance of rats. AB - AIM: To evaluate the influence of exogenous nitric oxides (NOx) on the antitumor resistance of rats, and to compare the activity of enzymes influencing the level of free radicals upon normal conditions and tumor growth. METHODS: The growth kinetics of Guerin carcinoma (GC) was studied. NO inhalation was performed in special chamber for 16 h per day during 1 month before GC transplantation. Proliferative activity of nonstimulated lymphocytes (PANSL), functional activity of peritoneal macrophages (PM) and cytotoxic activity of natural killer cells (NK) were studied in vitro. Proliferative activity of lymphocytes from lymph nodes (BTLR) was studied upon administration of Concanavalin A in vivo. The activity of xanthine oxidoreductase (XOR) and the level of lipid peroxidation (LPO) in tumor tissues were also evaluated. RESULTS: The inhalation of exogenous NOx results in toxic effect on the T-cells of immune system in vivo. Tumor growth was accompanied by activation of NK cells, PM and by decrease of proliferative activity of T-lymphocytes. The influence of NO accelerated the growth of tumor and was accompanied by the decrease of relative weight of thymus, peripheral lymph nodes and spleen 2.9, 2 and 1.5 fold, respectively; 4.9 fold increase of functional activity of PM; 2 and 2.3 fold decrease of PANSL and BTLR, respectively. In tumor tissue was observed 3.5 fold decrease of LPO level but the ratio of XOR isoforms increased by 18 fold mainly due to inactivation of xanthine dehydrogenase (XDH). CONCLUSION: The relation between the NO inhalation, immune status and antitumor resistance has been evaluated. The prolonged action of exogenous NOx negatively influence T-cells of immune system and caused hyperactivation of PM, sharp decrease of XDH activity and LPO level, and accompanied by accelerated tumor growth in vivo. PMID- 15812362 TI - MLL gene alterations in radiation-associated acute myeloid leukemia. AB - AIM: Although acute myelogenous leukemia (AML) arising after radiation exposure is considered to be secondary, little is known about the molecular mechanisms by which the radiation induces the leukemogenic phenotype. The aim of the study was to analyze whether the MLL translocations are as frequent in radiation-associated AML as in spontaneous AML cases. METHODS: Sixty one AML samples obtained at diagnosis were analyzed for the presence of MLL abnormalities using fluorescent in situ hybridization and/or reverse transcription polymerase chain reaction. Of these patients, 27 had experienced radiation exposure due to the Chernobyl accident, 32 were non-irradiated (spontaneous AML), and 2 developed therapy related AML after chemotherapy with topoisomerase II inhibitors. RESULTS: MLL gene translocations were detected in both groups of spontaneous and therapy related AML (1/32 and 1/2 cases respectively). The sole MLL rearrangement found in the group of radiation-associated AML patients was a duplication of the gene. CONCLUSION: Our data preclude the involvement of MLL gene translocations in radiation-induced leukemogenesis, but support the assumption that loss and gain of chromosomal material could be crucial in the leukemogenesis of AML patients with the history of radiation exposure due to the Chernobyl accident. PMID- 15812363 TI - Plasma level of lipid peroxidation, total sulphydryl groups and nitric oxide levels in cancer patients irradiated on different anatomic fields: a case-control study. AB - AIM: The aim of this study was to evaluate the effect of radiotherapy on lipid peroxidation, total sulfhydryl groups (RSH) and nitric oxide (NO radical) levels in plasma of cancer patients treated on different anatomic sites, and to compare the results with control subjects. METHODS: 89 patients with various malignancies and 33 healthy control subjects were recruited for the study. Cancer patients were grouped according to the irradiation portals (head and neck, thoracic, breast, abdominal and pelvic). Two blood samples before and after the completion of the fifth week of radiotherapy were collected from each patient. RESULTS: Plasma levels of malondialdehyde (MDA) and NO radical of cancer patients before irradiation were found higher than those in control subjects. After irradiation, a marked lipid peroxidation increase was demonstrated as well as the significant decrease of plasma RSH level. CONCLUSION: The alterations of the parameters indicate enhanced oxidant stress and point to the necessity of antioxidant prophylaxis upon application of radiotherapy. PMID- 15812364 TI - An in vitro study of cytotoxic effects of gossypol on human epidermoid larynx carcinoma cell line (HEp-2). AB - AIM: The study was aimed on an evaluation of the effect of gossypol at concentration of 25-150 microM on the level of apoptosis in the human epidermoid larynx carcinoma cell line, HEp-2, in vitro. METHODS: To determine the apoptotic effects of gossypol on growth of human epidermoid larynx carcinoma cell lines (HEp-2 cells), morphological evaluation of the cells and DNA fragmentation analysis were used. RESULTS: 6 h incubation of HEp-2 cells with gossypol at concentrations of 50 microM and 75 microM resulted in the increase of apoptosis rate by 26 and 23%, respectively. CONCLUSION: This preliminary study may constitute a base for in vivo studies of anticancer properties of gossypol. PMID- 15812365 TI - Where high-tech medicine was irrelevant. PMID- 15812366 TI - Screening the population for coronary artery disease: is it like screening for cancer? PMID- 15812367 TI - Role of BMIPP imaging for risk stratification in patients with coronary artery disease. PMID- 15812368 TI - Cardiovascular risk assessment in cancer patients undergoing major surgery. PMID- 15812369 TI - Screening for coronary heart disease: cardiology through the oncology looking glass. PMID- 15812370 TI - Pharmacokinetics and safety of binodenoson after intravenous dose escalation in healthy volunteers. AB - BACKGROUND: Binodenoson, a highly selective agonist of the adenosine A 2A receptor, is being developed as a short-acting coronary vasodilator as an adjunct to radiotracers for use in myocardial stress imaging. This study was designed to assess the single-dose pharmacokinetics, safety, and tolerability of intravenous binodenoson. METHODS AND RESULTS: This was a single-center, open-label, nonrandomized, dose-escalation study in 24 healthy volunteers. Each subject received 3 successive intravenous doses of binodenoson (0.1, 0.2, 0.4, 0.6, 1, 2, 3, 4, 5, and 6 microg/kg), each infused over a period of 10 minutes and separated by washout periods of at least 120 minutes. Generally, binodenoson was well tolerated. There were no serious adverse events. However, there was a dose related increase in adverse events (e.g., headache, nausea, vasodilation, chest pain), consistent with the pharmacology of the drug. Binodenoson exhibited linear pharmacokinetics as indicated by a dose-proportional increase in peak concentration (C max ) and area under the concentration-time curve (AUC). Systemic clearance was independent of dose but was correlated with body weight. The mean terminal half-life of binodenoson across all doses was 10 +/- 4 minutes. CONCLUSIONS: Overall, binodenoson was well tolerated and exhibited linear pharmacokinetics when administered intravenously over a 60-fold dose range from 0.1 to 6 microg/kg. PMID- 15812371 TI - Prognostic value of I-123 15-(p-iodophenyl)-3-(R,S) methylpentadecanoic acid myocardial imaging in patients with known or suspected coronary artery disease. AB - BACKGROUND: Although iodine 123 15-(p-iodophenyl)-3-(R , S) methylpentadecanoic acid (BMIPP) can assess abnormal utilization of fatty acid in the diseased myocardium, the prognostic value of BMIPP imaging at rest in patients with known or suspected coronary artery disease (CAD) remains unclear. METHODS AND RESULTS: A total of 270 patients were included by a retrospective search of the existing databases of 4 institutions. In addition to hard events, consisting of cardiac death and nonfatal myocardial infarction, any significant events including death, nonfatal myocardial infarction, coronary revascularization, heart failure, and unstable angina were assessed. During a median follow-up of 3.9 years, 33 patients had significant events, among whom 10 had hard events. Kaplan-Meier survival estimates revealed a hard event-free survival rate of 98% at 3 years in patients with a BMIPP defect score lower than 5 but 93% in those with a defect score of 5 or greater ( P = .03). With regard to significant events, the analysis showed an event-free survival rate of 92% at 3 years in patients with a BMIPP defect score lower than 5 but 80% in those with a defect score of 5 or greater ( P = .0003). CONCLUSIONS: These results indicate that resting BMIPP imaging has prognostic value and may have a role in the risk stratification of patients with known or suspected CAD. PMID- 15812372 TI - Tl-201 washout rate in remote normal regions in patients with prior myocardial infarction and left ventricular remodeling. AB - BACKGROUND: Myocardial characteristics of remote normal regions in patients with myocardial infarction (MI) and left ventricular (LV) remodeling have not been fully elucidated. Thus, we investigated this issue from the viewpoint of myocardial Tl-201 dynamics. METHODS AND RESULTS: In 14 patients with prior anterior MI, 10 with inferior MI, and 14 age-matched patients with atypical chest pain served as controls; exercise stress Tl-201 SPECT and cardiac catheterization were performed. Tl-201 washout rate was calculated for 8 myocardial segments, and LV end-diastolic volume index was obtained as a parameter of LV remodeling. LV end-diastolic volume index was greater in anterior MI patients than in control patients; in contrast, no significant difference was observed between inferior MI patients and control patients. The washout rate in remote normal regions was significantly less in anterior MI patients than in the corresponding segments in control patients (39.8% +/- 8.7% vs 48.4% +/- 4.4%, P < .01). There was no significant difference between inferior MI patients and control patients (43.6% +/- 6.9% vs 47.8% +/- 4.5%). CONCLUSIONS: Reduced Tl-201 washout rates in remote normal regions are found in patients with anterior MI and LV remodeling. Subclinical myocardial ischemia during exercise in remote normal regions exists and may be related to the pathologic condition of such LV walls. PMID- 15812373 TI - Comparative study of quantitative blood pool SPECT imaging with 180 degrees and 360 degrees acquisition orbits on accuracy of cardiac function. AB - BACKGROUND: Quantitative blood pool single photon emission computed tomography (SPECT) (QBS) can measure ejection fraction (EF) and volumes from gated blood pool single photon emission tomography (GBPS) working in fully automatic mode in 3-dimensional space. The effects of 180 degrees and 360 degrees data acquisition in GBPS have not been fully evaluated. This study compares the accuracy of 360 degrees and 180 degrees data acquisition for left ventricular (LV) systolic function in a clinical study and measures LV volume by GBPS compared with ultrasound echocardiography. METHODS AND RESULTS: The study population comprised 9 normal volunteers and 34 patients. GBPS data were acquired by use of 360 degrees rotation and 60 stops per head. All 60 (360 degrees ) and 30 (45 degrees right anterior oblique to 45 degrees left posterior oblique) pieces of projection data that were selected for reconstructing the 180 degrees data were reconstructed and both ventricular functional parameters were automatically obtained by QBS software. The contour of the LV septal wall was concave in 6 patients (14%) when processed at 180 degrees , whereas a concave septum at 360 degrees processing was observed in only 1 patient (2%). The coefficients of correlation between 180 degrees and 360 degrees were 0.467 for the end-diastolic volume (EDV) and 0.648 for the end-systolic volume (ESV). The mean 180 degrees EDV value (152.9 +/- 46.1 mL) was significantly smaller than that of the 360 degrees EDV (191 +/- 70.8 mL) ( P < .001). However, there was no significant difference between the 360 degrees EDV (0.623) and 180 degrees EDV (0.407) as compared by echocardiography ( P = .218). The agreement of the EF between both methods was close ( r = 0.894, P < .0001). The agreement of the right ventricular volumes between the 180 degrees and 360 degrees orbits was close ( r = 0.800 for EDV and 0.706 for ESV). The EF was relatively dispersed between the 180 degrees and 360 degrees methods ( r = 0.642). CONCLUSION: This study showed that SPECT image acquisition by use of both the 180 degrees method and the 360 degrees method considerably underestimated LV volume quantification. In addition, the LV volume with the 180 degrees method was significantly smaller than that with the 360 degrees method. Thus a 360 degrees acquisition orbit may be suitable for more quantitatively accurate results when blood pool imaging is performed with gated SPECT. PMID- 15812374 TI - Value of attenuation correction on ECG-gated SPECT myocardial perfusion imaging related to body mass index. AB - BACKGROUND: Obesity is a growing problem in the United States, and attenuation artifacts are more prevalent in this patient group. This study evaluated the impact of attenuation correction in patients with a high body mass index (BMI). METHODS AND RESULTS: Three readers interpreted gated attenuation-corrected and non-attenuation-corrected rest/stress technetium 99m sestamibi myocardial perfusion imaging results in 116 patients (BMI <30, n = 60; BMI > or =30, n = 56) who had coronary angiography no more than 60 days after imaging. Readers were blinded to all clinical information and as to whether myocardial perfusion imaging was attenuation-corrected or non-attenuation-corrected. Sensitivity, specificity, and accuracy for detection of coronary artery disease of 70% or greater for attenuation-corrected versus non-attenuation-corrected single photon emission computed tomography (SPECT) were 86% versus 89%, 79% versus 50%, and 84% versus 79%, respectively. Sensitivity, specificity, and accuracy for attenuation corrected versus non-attenuation-corrected SPECT for patients with BMI less than 30 were 90% versus 90%, 82% versus 64%, and 88% versus 85%, respectively. For BMI of 30 or greater, the results were 82% versus 87%, 76% versus 41%, and 80% versus 73%, respectively. There was a significant difference in specificity overall ( P = .02) and for the category of BMI of 30 or greater ( P = .03). CONCLUSIONS: This study demonstrates that electrocardiography-gated attenuation-corrected Tc-99m sestamibi SPECT myocardial perfusion imaging improves specificity compared with electrocardiography-gated non-attenuation-corrected SPECT myocardial perfusion imaging, especially in patients with BMI of 30 or greater. PMID- 15812375 TI - QRS score versus ST-segment changes in patients undergoing Tl-201 scintigraphy using dipyridamole infusion. AB - BACKGROUND: ST-segment changes after dipyridamole infusion followed by handgrip isometric stress lack diagnostic value, because of the low sensitivity for the detection of coronary artery disease (CAD). In addition, an abnormal QRS score during exercise had a greater diagnostic ability than ST-segment changes to detect CAD. This study was undertaken to compare QRS score values with ST-segment changes during thallium 201 scintigraphy via dipyridamole infusion. METHODS AND RESULTS: In this study 128 patients (101 men and 27 women), aged 53 to 72 years (mean, 59 +/- 8 years), underwent Tl-201 scintigraphy after dipyridamole infusion and handgrip isometric stress, as well as coronary angiography. QRS score values and ST-segment changes after dipyridamole infusion and handgrip isometric stress were also estimated. CAD was detected in 96 patients (75%), whereas normal coronary arteries were found in 32 (25%). According to scintigraphic data, 48 patients (37%) had no reversible perfusion defects whereas 80 (63%) had at least 1 reversible perfusion defect. Sensitivities for an abnormal QRS score and ST segment deviation were 68% versus 18% ( P < .01) for detection of CAD and 75% versus 19% for detection of myocardial ischemia ( P < .01), respectively. Similar specificities were found ( P = not significant). CONCLUSIONS: An abnormal QRS score significantly improves the low sensitivity of ST-segment changes for the detection of myocardial ischemia and CAD by use of Tl-201 scintigraphy with dipyridamole infusion and handgrip isometric stress. PMID- 15812376 TI - A historical perspective on measurement of ventricular function with scintigraphic techniques: Part II--Ventricular function with gated techniques for blood pool and perfusion imaging. PMID- 15812377 TI - Echocardiographic insights into regional flow-function relationships in coronary artery disease. PMID- 15812378 TI - Mimic ascites in a patient undergoing peritoneal dialysis on raw-data image Tc 99m tetrofosmin gated cardiac SPECT. PMID- 15812379 TI - High-resolution simultaneous imaging of SPECT, PET, and MRI tracers on histologic sections of myocardial infarction. PMID- 15812380 TI - Postischemic stunning in apical hypertrophic cardiomyopathy. PMID- 15812388 TI - What is quality in a transplant program? AB - Abstract Hematopoietic stem cell transplantation (HSCT) as a field of medicine has been subject to rapid development and evolution since its inception. Traditionally, HSCT has been used for therapy of a diverse group of malignancies, bone marrow failure states, and inherited disorders. The rapid evolution of transplantation technology coupled with the diverse outcomes associated with a heterogeneous group of patients has stymied the development of consensus over objective programmatic indicators of quality, especially as they pertain to outcomes. In some regard, the lack of consensus has caused transplant programs to respond to a more consumer-driven paradigm of evaluation. The community of providers of transplantation therapies has responded by establishing standards for accreditation of facilities and uniform presentation of programmatic data. Rapid acceptance of the need for meaningful quality programs to address all aspects of the transplant facility has moved HSCT to the forefront of implementing standards for medical practice. Because definition of optimal outcomes in HSCT is likely to remain elusive, it is imperative that providers involved with HSCT continue to take a leadership role in defining program quality through further research. PMID- 15812389 TI - Predictive utility of the attached segment in the quality control of a cord blood graft. AB - The limited number of progenitor stem cells in umbilical cord blood (UCB) enforces the optimization and strict control of all the procedures involved in its therapeutic use--ie, collection, processing, cryopreservation, thawing, and transportation--to ensure graft potency at transplantation. For this reason, international UCB standards recommend storage of a cell sample attached to the UCB unit as a quantitative and functional control of the unit selected for transplantation. To validate the use of the sample attached to the UCB unit as a quality-control tool for the final product, UCB units (n = 20) stored in liquid nitrogen with the Bioarchive system were analyzed. The UCB units and their attached segments were thawed, and the number and viability of total nucleated cells, mononucleated cells, CD45 + cells, and CD34+ cells were determined, as were colony-forming cell counts. There was no significant difference between UCB units and segments for any of the parameters assessed. Additionally, the linear correlation coefficient (R2) in these paired samples was 0.85 and 0.78 for CD34+ cells and colony-forming cells, respectively. In conclusion, the cell sample in the tube segment physically linked to the transplant UCB bag predicts the total cell content and functionality of the unit and may serve as a source for final quality control of the UCB unit before transplantation. PMID- 15812381 TI - Targeting the vulnerable plaque: the evolving role of nuclear imaging. AB - The majority of acute ischemic events relating to atherosclerosis are caused by plaque rupture and ensuing thrombosis. The risk of plaque rupture is dictated in part by plaque morphology, which in turn is influenced by pathophysiologic mechanisms at the cellular and molecular level. Anatomic imaging modalities such as intravascular ultrasound, high-resolution magnetic resonance imaging, and multislice computed tomography can identify morphologic features of the vulnerable plaque, such as a large lipid core and thin fibrous cap, but give little or no information regarding molecular and cellular mechanisms, such as endothelial function, macrophage activation, lipid transport and metabolism, and cell death. Recent studies suggest that nuclear imaging may be able to provide images of sufficient quality to identify and quantify some of these molecular and cellular pathophysiologic processes. In the future this could allow for the early identification and noninvasive monitoring of vulnerable plaque. PMID- 15812390 TI - Lack of correlation between an assay used to determine early marrow allograft rejection and long-term chimerism after murine allogeneic bone marrow transplantation: effects of marrow dose. AB - The acute rejection of bone marrow (BM) allografts by host effectors can occur within a short period after BM transplantation (BMT) in lethally irradiated mice. Common assays used to ascertain engraftment/resistance involve measuring the growth of granulocyte/monocyte progenitors (colony-forming unit-granulocyte macrophage) in vitro or splenocyte proliferation assessed by radioisotope incorporation in vivo 5 to 8 days after BMT. However, the correlation of the long term outcome of BMT with the kinetics of recovery by using the dose of allogeneic BM cells (BMCs) that leads to early rejection as determined by the in vitro assessment has not been extensively studied. Thus, to investigate whether the early rejection of donor BMCs is an indication of a long-term engraftment failure, C57BL/6 (H2b) mice were lethally irradiated and transplanted with various doses of BALB/c (H2d) BMCs. The short-term engraftment of donor precursors (colony-forming unit-granulocyte-macrophage), the kinetics of hematopoietic cell recovery, the extent of donor chimerism, and the proportion of the recipients with long-term survival were determined. The results show that the kinetics and extent of hematopoietic cell recovery were significantly delayed in mice receiving limiting doses of BMCs that were rejected or severely resisted at day 8 after BMT. However, a proportion of these mice survived up to 98 days after BMT with mixed chimerism or donor chimerism. This study demonstrates that early rejection of BM precursors, as assessed by measurement of myeloid progenitors in the spleen after BMT, does not always correlate with the long-term outcome of the marrow allograft and that significant variability is inherent in the extent of chimerism when threshold amounts of BMCs are used. PMID- 15812391 TI - CD8 T-cell ability to exert immunodomination correlates with T-cell receptor: epitope association rate. AB - When presented alone, H7 a and HY antigens elicit CD8 T-cell responses of similar amplitude, but H7 a totally abrogates the response to HY when both antigens are presented on the same antigen-presenting cell. We found that H7a- and HY-specific T-cell precursors had similar frequencies in nonimmune mice and expressed similar levels of CD5. The H7a -specific CD8 T-cell repertoire harvested at the time of primary response showed highly restricted T-cell receptor (TCR) diversity. Furthermore, T cells specific for H7a and HY expressed equivalent levels of CD8 and TCR and displayed similar tetramer decay rates. The key difference was that anti-H7a T cells exhibited a much more rapid TCR:epitope on-rate than anti-HY T cells. Coupled with evidence that primed CD8 T cells limit the duration of antigen presentation by killing or inactivating antigen-presenting cells, our data support a novel and simple model for immunodomination: the main feature of T cells that exert immunodomination is that, compared with other T cells, they are functionally primed after a shorter duration of antigen presentation. PMID- 15812392 TI - HLA-matched unrelated donor hematopoietic cell transplantation after nonmyeloablative conditioning for patients with chronic myeloid leukemia. AB - We evaluated 10/10 HLA antigen-matched unrelated hematopoietic cell transplantation (HCT) after nonmyeloablative conditioning with fludarabine 3 x 30 mg/m2 and 2 Gy of total body irradiation as treatment for patients with chronic myeloid leukemia who were ineligible for conventional HCT. Data from 21 consecutive patients in first chronic phase (CP1; n = 12), accelerated phase (AP; n = 5), second CP (CP2; n = 3), and blast crisis (n = 1) were analyzed. Stem cell sources were bone marrow (n = 4) or granulocyte colony-stimulating factor mobilized peripheral blood mononuclear cells (G-PBMCs; n = 17). The patient who underwent transplantation in blast crisis died on day 21 (too early to be evaluated for engraftment) from progressive disease. Sustained engraftment was achieved in 5 of 12 patients who underwent transplantation in CP1, 4 of 5 patients who underwent transplantation in AP, and 2 of 3 patients who underwent transplantation in CP2, whereas 9 patients rejected their grafts between 28 and 400 days after HCT. Specifically, 1 of 4 marrow recipients and 10 of 17 G-PBMC recipients achieved sustained engraftment. Graft rejections were nonfatal in all cases and were followed by autologous reconstitution with persistence or recurrence of chronic myeloid leukemia. Seven of 11 patients with sustained engraftment--including all 5 patients in CP1, 2 of 4 patients in AP, and neither of the 2 patients in CP2--were alive in complete cytogenetic remissions 118 to 1205 days (median, 867 days) after HCT. Two of the remaining 4 patients died of nonrelapse causes in complete (n = 1) or major (n = 1) cytogenetic remissions, and 2 died of progressive disease. Further efforts are directed at reducing the risk of graft rejection by exclusive use of G-PBMC and increasing the degree of pretransplantation immunosuppression. PMID- 15812393 TI - Increased gene expression of chemokine receptors is correlated with acute graft versus-host disease after allogeneic stem cell transplantation. AB - Acute graft-versus-host disease (GVHD) is still a major complication after allogeneic stem cell transplantation. It is initiated by infiltrating donor T cells specific against the host antigens. Because T-cell migration is largely controlled by the expression of chemokines and chemokine receptors, we investigated the relation of acute GVHD and chemokine receptor expression in peripheral blood in 50 patients after allogeneic stem cell transplantation. The gene expression of the chemokine receptors CCR1, CCR2, CCR5, and CXCR3 was monitored by using quantitative real-time polymerase chain reaction. Among the 36 patients diagnosed with acute GVHD, 10 developed a second episode of acute GVHD. Therefore, gene-expression levels could be analyzed in 46 occasions of acute GVHD. When all 4 markers were evaluated at the same time, increased gene expression levels of at least 1 of the 4 markers were seen in 44 of 46 episodes of acute GVHD. The median increase of the 4 markers ranged from 3x to 12x in connection with acute GVHD. It is interesting to note that we saw increasing gene expression levels a few days before acute GVHD was diagnosed clinically at 17, 15, 22, and 19 occasions for CCR5, CXCR3, CCR1, and CCR2, respectively. The median number of days before diagnosis ranged from 3 to 5. Although they are not specific for acute GVHD, quantitative monitoring of the gene expression of chemokine receptors may be a valuable molecular method to monitor and diagnose acute GVHD. PMID- 15812394 TI - Comparison of lung function after myeloablative and 2 Gy of total body irradiation-based regimens for hematopoietic stem cell transplantation. AB - Lung function decline is a well-recognized occurrence after myeloablative hematopoietic stem cell transplantation (HCT) that has not been studied after nonmyeloablative conditioning regimens. We examined the lung function of patients before and after 2-Gy total body irradiation-based nonmyeloablative and myeloablative preparative regimens. Before HCT, at day 100, and 1 year after HCT, nonmyeloablative patients had lower 1-second forced expiratory volume (FEV1), forced vital capacity, total lung capacity, residual volume, and carbon monoxide diffusion capacity. However, after transplantation, the risk for experiencing a >20% per year decrease of FEV 1 was significantly lower for nonmyeloablative than myeloablative patients >50 years of age (odds ratio, 0.3; 95% confidence interval, 0.1-0.8; P = .01). Lower pretransplantation FEV 1 was associated with a higher mortality rate for both groups, with the highest mortality risk among patients with a pretransplantation FEV 1 <60% (nonmyeloablative: hazard ratio, 3.9; 95% confidence interval, 1.9-8.0; myeloablative: hazard ratio, 7.2; 95% confidence interval, 2.5-21.2). These results suggest that despite having worse lung function, patients who receive the 2-Gy total body irradiation-based nonmyeloablative regimen will likely experience less pulmonary toxicity than patients who receive a myeloablative regimen, and this may have important clinical implications when deciding on a conditioning regimen for patients >50 years of age with compromised pretransplantation lung function. PMID- 15812395 TI - Phase I and pharmacokinetic study of docetaxel combined with melphalan and carboplatin, with autologous hematopoietic progenitor cell support, in patients with advanced refractory malignancies. AB - The purpose of this study was to define the maximal tolerated dose (MTD), extramedullary toxicities, and pharmacokinetics of docetaxel combined with high dose melphalan and carboplatin with autologous hematopoietic progenitor cell support. Fifty-nine patients with advanced refractory malignancy (32 breast cancer, 10 non-Hodgkin lymphoma, 6 germ cell tumors, 4 Hodgkin disease, 4 ovarian cancer, 2 sarcoma, and 1 unknown primary adenocarcinoma) with a median of 3 prior chemotherapy regimens and a median of 3 organs involved were enrolled. Treatment included docetaxel (150-550 mg/m2 infused over 2 hours on day -6), melphalan (150 165 mg/m2 infused over 15 minutes from day -5 to -3), and carboplatin (1000-1300 mg/m2 as a 72-hour continuous infusion from day -5). Five patients died from direct regimen-related organ toxicity (2 capillary leak syndrome, 2 enterocolitis, and 1 hepatic toxicity), and 1 additional patient died from pulmonary aspergillosis. The docetaxel MTD was defined as 400 mg/m 2 , combined with melphalan (150 mg/m2 ) and carboplatin (1000 mg/m2 ). The MTD cohort was expanded to enroll a total of 26 patients, 1 of whom died from toxic enterocolitis. The remaining 25 patients presented the following extramedullary toxicity profile, which was manageable and largely reversible: stomatitis, myoarthralgias, peripheral neuropathy, gastrointestinal and cutaneous toxicities, and syndrome of inappropriate antidiuretic hormone secretion. Docetaxel exhibited linear pharmacokinetics in the dose range tested (150-550 mg/m2 ). Pharmacodynamic correlations were noted between the docetaxel area under the curve and peripheral neuropathy or stomatitis. The response rate among 38 patients with measurable disease was 95%, with 47% complete responses. At a median follow-up of 26 months (range, 7-72 months), the 3-year event-free survival and overall survival were 26% and 36%, respectively. In conclusion, a 4 fold dose escalation of docetaxel, combined with melphalan and carboplatin, is feasible with autologous hematopoietic progenitor cell support. The notable activity of this regimen in treatment-refractory patients warrants its further evaluation. PMID- 15812396 TI - Efficacy of mycophenolate mofetil in the treatment of chronic graft-versus-host disease. AB - Current treatment of chronic graft-versus-host disease (cGVHD) with prednisone (PSE) alone or with added cyclosporine or tacrolimus still has a very high failure and complication rate, and new treatment approaches are needed for both primary and salvage therapy. Mycophenolate mofetil (MMF) is an immunosuppressive agent currently in use for acute graft-versus-host disease prophylaxis. To determine whether MMF had activity in the treatment of cGVHD, we added MMF to standard cyclosporine, tacrolimus, and/or PSE as salvage/second-line (n = 24) or first-line (n = 10) therapy in 34 patients. Nine (90%) of 10 patients receiving first-line and 18 (75%) of 24 receiving second-line MMF therapy responded. Twelve (35%) patients had a complete remission, 15 (44%) had a partial remission, 5 (15%) had stable disease, and only 2 (6%) had progressive disease. Out of 30 patients receiving PSE, 22 (73%) were able to decrease PSE doses (median decrease of 50%; range, 25%-100%). With a median follow-up of 24 months (range, 6-28 months), 29 (85%) patients are alive. Three patients had to discontinue MMF because of abdominal cramps within 3 months of starting treatment. These data suggest that MMF is an active, well-tolerated agent in the treatment of cGVHD and may have a beneficial effect on the survival of patients with this complication. PMID- 15812397 TI - Reduced-intensity unrelated cord blood transplantation for patients with advanced malignant lymphoma. AB - We report the results of reduced-intensity unrelated cord blood transplantation (RI-UCBT) in patients with advanced malignant lymphoma. Twenty patients (median age, 46.5 years; range, 27-66 years) underwent RI-UCBT with a preparative regimen consisting of fludarabine 125 mg/m2 , melphalan 80 mg/m 2 , and 4 Gy of total body irradiation. The median infused total cell dose was 2.75 x 10(7)/kg (range, 2.3-3.4 x 10(7)/kg). Graft-versus-host disease (GVHD) prophylaxis was composed of cyclosporine or tacrolimus alone. Fifteen patients achieved primary neutrophil engraftment after a median of 20 days. Eight patients developed grade II to IV acute GVHD, and 2 developed chronic GVHD. Of the 16 patients with evaluable disease, 10 achieved a complete response. Primary disease recurred in 1 patient, and transplant-related mortality within 100 days occurred in 8 of 20 patients. The estimated 1-year probability of progression-free survival was 50%. These data suggest that RI-UCBT is a feasible option for patients with refractory lymphoma who lack an HLA-matched donor. PMID- 15812400 TI - ASGE guideline: guidelines for credentialing and granting privileges for capsule endoscopy. PMID- 15812401 TI - Endoscopic treatment of high-grade dysplasia and early stage cancer in Barrett's esophagus. AB - BACKGROUND: The aim of this study was to prospectively evaluate endoscopic resection (ER) combined with photodynamic therapy (PDT) for the treatment of selected patients with early neoplasia in Barrett's esophagus. METHODS: Patients with Barrett's esophagus and neoplastic lesions <2 cm in diameter and no sign of submucosal infiltration, positive lymph nodes, or distant metastasis underwent diagnostic ER (cap technique). Patients with a T1sm tumor in the resection specimen were referred for surgery; those with a T1m or a less invasive tumor underwent additional endoscopic therapy (ER, PDT, and/or argon plasma coagulation [APC]), or they were followed. PDT was performed with 5-aminolevulinic acid and a light dose of 100 J/cm 2 at lambda = 632 nm. RESULTS: Thirty-three patients underwent diagnostic ER. Endoscopic treatment was not performed in 5 patients, who underwent surgery (4 T1sm; 1, patient preference). Five patients were immediately entered into a follow-up protocol, and 23 received additional endoscopic treatment (13 additional ER, 19 PDT, 3 APC). Endoscopic treatment was successful in 26/28 patients; no severe complication was observed. During follow up (median 19 months, range 13-24 months), 5/26 patients had a recurrence of high grade dysplasia: all were successfully re-treated with ER. At the end of follow up, 26/33 originally enrolled patients (79%) and 26/28 endoscopically treated patients (93%) were in local remission. CONCLUSIONS: Endoscopic therapy is safe and effective for selected patients with early stage neoplasia in Barrett's esophagus. PMID- 15812402 TI - Visible endoscopic and histologic changes in the cardia, before and after complete Barrett's esophagus ablation. AB - BACKGROUND: Adverse events associated with the thermal ablation of Barrett's esophagus (BE) include the generation of gastric mucosa buried beneath the neosquamous regrowth, and unrecognized development and growth of adenocarcinomas. No reports exist regarding the endoscopic appearance and histology of the cardia before and after BE ablation. The aim of our study was to assess the relative frequency of the occurrence of visible endoscopic and histologic changes in the cardia, before and after complete BE ablation. METHODS: A subset analysis of patients with uncomplicated BE, BE with dysplasia, or early carcinoma, who had been enrolled into one of 4 ongoing prospective studies of mucosal ablation, was examined. Eighty-two patients were identified who entered a BE ablation study, with 75 of these completing BE mucosal ablation. Cardia biopsy specimens were taken in all patients before ablation and serially after BE ablation. Cardia histology was graded by using the modified Sydney System for gastritis. RESULTS: Before ablation, cardia nodules were noted in 3, cardia intestinal metaplasia (IM) in 7 (8.5%), and none harbored cardia dysplasia. Postablation surveillance ranged from 3 to 75 months (mean 31.1 months [19.5]). Six subjects (8%) developed cardia nodules during surveillance; cardia IM was found in 21(28%), with 17 of these being a new finding (incidence of 25%). Cardia low-grade dysplasia incidence was 1.3% and high-grade dysplasia was 4% after BE ablation. CONCLUSIONS: The pathophysiology of the abnormal cardia histology and the endoscopic lesions (nodules) is unclear, but endoscopic surveillance of not only the neosquamous epithelium but also the cardia should be considered after ablation, especially in those high-grade dysplasia and early adenocarcinoma BE patients. PMID- 15812403 TI - Efficacy of an overtube for reducing the risk of peristomal infection after PEG placement: a prospective, randomized comparison study. AB - BACKGROUND: With the conventional pull method of PEG placement, there is a significant risk of wound infection from contamination of the gastrostomy catheter as it passes through the oral cavity. This study compared the occurrence of peristomal wound infection associated with PEG placement with and without use of an overtube. METHODS: Consecutive patients with dysphagia were randomized to undergo PEG placement with (Group I) or without (Group II) an overtube. For each patient, the peristomal area was evaluated daily for 1 week after PEG placement. The presence of erythema and of exudate were scored on a scale of 0 to 4; induration was scored on a scale of 0 to 3. Criteria for infection were a maximum combined score of 8 or higher, or the presence of microscopic and microbiologic evidence of suppurating exudate. In each group, cefazolin was administered prophylactically (2 g/d intravenously) for 3 days. For patients who had received an antibiotic(s) before PEG placement, the same antibiotic(s) was used. All procedures in both groups were performed by one of two investigators who used the pull method. RESULTS: A total of 76 patients were randomized; 3 were excluded from analysis, because death occurred within 1 week after the procedure. Two of 3 deaths were procedure-related (aspiration pneumonia in Group I, peritonitis in Group II). Data for 37 patients in Group I and 36 in Group II were analyzed. There was no significant difference between the groups with respect to baseline characteristics. The occurrence of peristomal infection within 1 week of PEG was significantly lower in Group I compared with Group II (2 vs. 12; p = 0.0029). The mean daily combined scores in Group I also were significantly lower than those in Group II ( p < 0.0001), and the median maximum parameter scores in Group I were significantly lower than those in Group II (erythema, p = 0.0062; induration, p = 0.0390; exudate, p < 0.0001), although the nominal significance for induration was removed by correction for the multiple testing of data. One patient excluded from Group II died from sepsis because of procedure-induced peritonitis. Among the 73 enrolled patients, there was no procedure-related mortality or clinically important wound infections that required surgical intervention in either group. CONCLUSIONS: Use of an overtube during PEG placement reduces the risk of peristomal wound infection. PMID- 15812404 TI - Efficacy and complications of covered Wallstents in malignant distal biliary obstruction. AB - BACKGROUND: This study evaluated the efficacy and the complications associated with the use of the covered Wallstent in the setting of unresectable malignant biliary obstruction. METHODS: Between March 2001 and January 2003, all patients with distal malignant biliary obstruction that required drainage were treated with a covered Wallstent. Every 2 months, the patients were evaluated clinically and biochemical tests of liver function were obtained. Data were recorded for the following variables: early complications (within 30 days of stent placement), early and late stent occlusion, duration of stent patency, need for subsequent biliary intervention, and patient survival. RESULTS: A total of 88 covered Wallstents were inserted in 80 patients. Stent patency rates at 3, 6, and 12 months were 90%, 82%, and 78%, respectively. Complications included stent migration (5), stent occlusion (12), episodes of cholecystitis (3), and episodes of post-ERCP pancreatitis (5). Biliary intervention was required in 9 patients subsequent to placement of the initial covered Wallstent. CONCLUSIONS: Deployment of a covered Wallstent is safe and relatively easy. It achieves biliary drainage with an acceptable risk to benefit ratio in the majority of patients with distal malignant biliary obstruction. PMID- 15812405 TI - Metal stents for distal biliary malignancy: have we got you covered? PMID- 15812406 TI - Correlation of endoscopic optical coherence tomography with histology in the lower-GI tract. AB - BACKGROUND: Optical coherence tomography (OCT), a noninvasive optical imaging technique, provides high-resolution cross-sectional images of tissue microstructure. We developed a system for real-time endoscopic OCT (EOCT) of the human GI tract. During clinical trials, the structure of mucosa and submucosa, glands, blood vessels, pits, villi, and crypts was observed in a range of GI organs. Although EOCT images are thought to accurately depict actual histologic features, there are few data to support this assumption. Therefore, the present study correlated images acquired with an EOCT imaging system in vitro to corresponding histologic sections. METHODS: EOCT images were obtained of fresh specimens of ileum, colon, and rectum that then were fixed in formalin and were processed for microscopic evaluation by using standard methods. The thickness of mucosa and of submucosa was determined for both EOCT images and histologic slides. RESULTS: The first hyper-reflective layer in the EOCT images was identified as mucosa. A close correlation (R 2=0.84) was observed between EOCT and histology. Furthermore, the submucosa and the muscularis propria could be identified as the next deepest hyporeflective band and a hyper-reflective layer, respectively, in EOCT images. The submucosa was found to be more compressible than mucosa, and its EOCT appearance was dependent on its content of adipose tissue. CONCLUSIONS: EOCT provides images that precisely correlate with the histologic structure of the mucosa and the submucosa of the GI tract. PMID- 15812407 TI - A prospective study of the frequency and the topographical distribution of colon neoplasia in asymptomatic average-risk Chinese adults as determined by colonoscopic screening. AB - BACKGROUND: The goal of this study was to determine the frequency of colorectal neoplasia in an asymptomatic Taiwanese population and the topographic distribution of lesions relative to age and gender. METHODS: Colonoscopy was performed in 1846 consecutive asymptomatic adults undergoing a health evaluation in 2003. Neoplastic lesions were considered advanced if they exceeded 10 mm in size, had a villous component, or contained moderately or severely dysplastic tissue or invasive cancer. Lesions at and proximal to the splenic flexure were considered proximal in location; those distal to the splenic flexure were classified as distal in location. RESULTS: Of 1741 (94.3%) patients (1041 men, 700 women; mean 52.5 years) enrolled, 1708 (98.1%) underwent total colonoscopy. Of these patients, 263 (15.4%) had colorectal neoplasia; 51 (3.0%) had advanced lesions. A total of 331 lesions were detected; 125 (37.8%) were proximal in location. Two thirds of patients with proximal advanced lesions (66.7%, 10/15) had no distal lesion. The proportion of patients with proximal or proximal plus distal lesions increased with age ( p = 0.027). CONCLUSIONS: Colonoscopy is an effective primary screening modality for colorectal neoplasia in asymptomatic Chinese patients. Many lesions would be missed, especially in the elderly, if only sigmoidoscopy was used for initial screening. PMID- 15812408 TI - Assessment of the impact of an educational course on knowledge of appropriate EUS indications. AB - BACKGROUND: The knowledge level of EUS among gastroenterologists likely influences the appropriateness of requested indications for EUS. It remains unknown what the impact is of a short EUS course, involving didactic teaching, on knowledge levels of EUS indications for EUS. The aim of this study was to assess the impact of a 3-day educational course on knowledge levels of attending gastroenterologists regarding the appropriateness of indications for EUS. METHODS: A questionnaire was designed that tested knowledge of indications for EUS in 4 anatomic sites: esophagus, gastroduodenum, hepatopancreatobiliary system, and colorectum. This questionnaire was distributed to all attendees of a 3-day EUS educational course. All attendees completed the survey before and immediately after the course. RESULTS: A total of 24 gastroenterologists completed the pre- and post-course survey. Before the course, respondents scored highest in questions on EUS applications in the gastroduodenum (94%) and the hepatopancreatobiliary system (88%) compared with the esophagus (72%) and the colorectum (74%). Statistically significant improvements in knowledge were recorded in all organ categories: gastroduodenum (100%, p = 0.002 vs. pretest score), hepatopancreatobiliary system (99%, p < 0.0001), esophagus (92%, p < 0.0001), and colorectum (93%, p = 0.0004). The biggest improvement was observed in knowledge levels for the esophagus (20%) and the colorectum (18%). CONCLUSIONS: There was a consistent improvement in the gastroenterologists' knowledge levels of EUS indications among all organ categories after an educational course. Our findings suggest that education enhances gastroenterologists' understanding of EUS. Future studies should seek to assess the impact of these improved knowledge levels on the appropriateness of EUS referral patterns. PMID- 15812409 TI - Fibrin glue, healing of gastric mucosal injury, and expression of growth factors: results from a human in vivo study. AB - BACKGROUND: Fibrin glue is used in the endoscopic therapy of bleeding ulcerations. Accelerated closure of ulcers has been attributed to this treatment; the biologic reason, however, remains unclear. METHODS: Two artificial gastric lesions were induced in healthy, Helicobacter pylori negative volunteers and were treated by injection of either saline solution or fibrin glue. After 72 hours, resulting ulcers were measured and biopsy specimens were taken for immunohistochemistry (to identify proliferating cells and small vessels) and assessment of growth factor messenger RNA (mRNA) expression (platelet derived growth factor, vascular endothelial growth factor, fibroblast growth factor 2 [FGF-2]) by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: After 72 hours, most lesions exposed to fibrin glue were smaller than the corresponding ones treated with saline solution. The ulcer rim was more pronounced; immunohistochemistry revealed more proliferating cells (p < 0.02 compared with saline solution). The number of microvessels also increased, though this difference did not reach statistical significance (p = 0.10). FGF-2 mRNA expression markedly increased (about 7-fold compared with the control [ p < 0.001], and about 5-fold compared with saline solution [ p < 0.015]); whereas, with respect to platelet derived growth factor and vascular endothelial growth factor mRNAs, only small changes occurred. CONCLUSIONS: Fibrin glue positively modulates gastric ulcer healing by causing an increase in the number of proliferating cells in the ulcer margin and also possibly enhances the density of microvessels. These changes are accompanied by an enhanced expression of FGF-2, which is known to exert beneficial effects on ulcer healing. PMID- 15812410 TI - Predictors of outcome in pancreatic duct disruption managed by endoscopic transpapillary stent placement. AB - BACKGROUND: The aim of this study was to identify predictors of outcome for pancreatic duct (PD) disruption managed by endoscopic transpapillary stent insertion. METHODS: An endoscopy database was used to identify patients with PD disruption, defined as extravasation of contrast from the PD during ERCP. Data collected included demographic information, the results of imaging studies, management before endoscopic intervention, and outcomes after stent placement. Stents typically were exchanged at intervals of 6 to 8 weeks. Success was defined as clinical and pancreatographic resolution of the PD disruption. RESULTS: Ninety seven consecutive patients (34 women, 63 men; mean age, 53.7 [12.3] years) with PD disruption seen from 1995 to 2002 were identified. Causes of the disruption were the following: chronic pancreatitis (47), acute pancreatitis (44), operative injury (4), and trauma (2). Transpapillary PD stent insertion was technically successful in 92 (95%) patients; two underwent a combined cystenterostomy. The median duration of stent placement was 58 days (range 4-640 days). The outcome of stent insertion was successful in 52 patients (55%), unsuccessful in 32 (36%), and indeterminate in 8 (9%). On univariate analysis, a partially disrupted PD (p < 0.001), a disruption in the body of the pancreas (p = 0.04), a stent positioned to bridge the disruption (p < 0.001), and a longer duration of stent therapy (p = 0.03) were associated with a successful outcome. On multivariable logistic regression, only a partially disrupted duct and a stent bridging the disruption correlated with a successful outcome. Complications occurred in 6 patients. CONCLUSIONS: Successful resolution of PD disruption by transpapillary stent insertion depends on the type of disruption and the ability to bridge the disrupted duct with a stent. PMID- 15812411 TI - Efficacy of main pancreatic-duct endoscopic drainage in patients with chronic pancreatitis, continuous pain, and dilated duct. AB - BACKGROUND: The aim of endoscopic treatment in patients with chronic pancreatitis is to achieve decompression of the pancreatic duct, because duct obstruction with increased pressure within the duct is one of the leading causes of pain in these patients. The majority of patients suffer from relapsing pain, thus making it difficult to evaluate the efficacy of therapy. The outcome of endoscopic treatment on pain has been evaluated in patients with continuous pain (present for more than 1 month, at least 5 days per week, requiring daily analgesic therapy) and dilated duct. METHODS: Of 343 patients who underwent endotherapy for chronic pancreatitis in a 15-year period, 22 (6.4%)(19 men, mean age 48 years, alcohol abuse 14) had continuous pain and a dilated pancreatic duct. RESULTS: Endotherapy was successful in all patients, with no procedure-related mortality and only mild complications. Pain disappeared, and analgesics could be discontinued in all patients immediately after endotherapy. Six patients are pain free after a mean period of 5.5 years. One pain-free patient died after 3.4 years from myocardial infarction. Five patients were successfully endoscopically re treated for pain relapses. Four patients underwent surgery for frequent pain relapses after a mean period of 2.5 years. Six patients were lost to follow-up. CONCLUSIONS: Endotherapy should be considered as the initial treatment of choice in patients with chronic pancreatitis, dilated duct, and continuous pain. PMID- 15812412 TI - Endoscopic therapy for pain in chronic pancreatitis: is it time for the naysayers to throw in the towel? PMID- 15812413 TI - The pathway to academic success starts during fellowship. PMID- 15812415 TI - Bleeding stromal tumor. PMID- 15812416 TI - Removal of a gastroplasty ring. PMID- 15812417 TI - Gastric stump carcinoma. PMID- 15812418 TI - Biliary fascioliasis. PMID- 15812419 TI - Endoscopic pyloromyotomy for congenital pyloric stenosis. AB - BACKGROUND: The advent of sophisticated endoscopic devices allows for a variety of procedures heretofore performed surgically. This study describes the results of endoscopic pyloromyotomy for congenital hypertrophic pyloric stenosis (CHPS). METHODS: Ten consecutive infants (7 boys, 3 girls; age range 3-7 weeks), with a diagnosis of CHPS, underwent endoscopic pyloromyotomy while under conscious sedation by using an endoscopic electrosurgical needle knife or a sphincterotome. Incisions were made from the antral to the duodenal side of the pylorus. All procedures except one were performed on an outpatient basis. RESULTS: No complication was encountered in any patient. All patients began regular feedings as soon as they recovered from the effects of the sedative medication and were discharged on the same day. At follow-up (range 6 months to 2 years), all patients were doing well. CONCLUSIONS: Surgery for CHPS has undergone little change in recent decades except for the advent of the laparoscopic approach. Surgical pyloromyotomy is considered simple, inexpensive, and safe. Endoscopic pyloromyotomy is equally simple, probably less expensive, and it can be performed as an outpatient procedure. PMID- 15812420 TI - Experimental studies of transgastric gallbladder surgery: cholecystectomy and cholecystogastric anastomosis (videos). AB - BACKGROUND: Transgastric flexible endoscopic surgery might offer advantages over open and laparoscopic surgery. The aim of this study was to develop methods for performing transgastric biliary endosurgery. METHODS: Cholecystectomies and biliary anastomoses were performed in 8 anesthetized pigs (27-30 kg) in nonsurvival studies. Two endoscopes passed perorally were inserted through the stomach wall after needle-knife incision. Endoscope-induced pneumoperitoneum allowed viewing and manipulation of the gallbladder with both endoscopes independently. The cystic duct was dissected, clipped, and transected. Cholecystectomy was performed with one of two methods: either by using two endoscopes, or a single endoscope and a 5-mm-diameter grasping instrument inserted transabdominally. Clips and sutures were used to attach the gallbladder to the stomach wall, and an incision was made to form a cholecystogastrostomy. In survival experiments in 8 pigs, transgastric incisions were closed with endoscopic sutures. RESULTS: The gallbladder was successfully removed in 8 pigs (nonsurvival experiments). The time for the procedure ranged from 2.5 hours to 40 minutes and decreased with experience. At postmortem examination, clips placed on the cystic duct and the artery were secure. An anastomosis was successfully formed between gallbladder and stomach in 3 pigs. In 8 pigs, full-thickness incisions in the stomach wall were closed with two to 4 stitches. All 8 pigs survived (median follow-up, 22 days; range 14-28 days). CONCLUSIONS: Transgastric gallbladder surgery, including cholecystectomy and biliary anastomosis, is feasible. Full-thickness gastric incisions were safely closed in survival studies. The efficacy and the safety of transgastric surgery merits further study. PMID- 15812421 TI - Characterization of secondary GI lesions with anaplastic large-cell (Ki-1) lymphoma: a first report of two cases. PMID- 15812422 TI - A first report of tumor seeding because of EUS-guided FNA of a pancreatic adenocarcinoma. PMID- 15812423 TI - Acute proctosigmoiditis caused by a dexamethasone mouthwash enema. PMID- 15812424 TI - Asymptomatic esophageal perforation after foreign body ingestion. PMID- 15812425 TI - Cerebral air embolism during upper endoscopy: case report and review. PMID- 15812426 TI - Endoscopic incision of a refractory esophageal stricture: novel management with an endoscopic scissors. PMID- 15812428 TI - Bleeding polyp in the mid small intestine identified by capsule endoscopy and treated by double-balloon endoscopy. PMID- 15812427 TI - Neoadjuvant therapy with imatinib mesylate for locally advanced GI stromal tumor. PMID- 15812429 TI - Gastric ulceration and perforation as a complication of a left ventricular assist device. PMID- 15812430 TI - Watermelon colon treated by argon plasma coagulation. PMID- 15812431 TI - Colocutaneous fistula caused by a self-expanding metallic stent. PMID- 15812432 TI - Atypical EUS features of gastric inflammatory fibroid polyps. PMID- 15812433 TI - Pancreas divisum as the etiology of acute pancreatitis in children. PMID- 15812434 TI - Other factors affecting patient tolerance and acceptance of unsedated upper-GI endoscopy. PMID- 15812435 TI - Indications for ERCP in children with recurrent pancreatitis: predictive value of abdominal US and serum amylase levels. PMID- 15812436 TI - Should adolescents be paid to participate in research? PMID- 15812437 TI - The impact of chronic disease. PMID- 15812438 TI - Severe BPD is decreasing. PMID- 15812439 TI - Adenovirus and ophthalmologic examinations. PMID- 15812440 TI - Exploring risks for sexually transmitted infections. PMID- 15812441 TI - What is the best method to identify cardiovascular risk related to obesity? PMID- 15812443 TI - Adenovirus in the neonatal intensive care unit: formidable, forgotten foe. PMID- 15812444 TI - Dyspepsia in adolescence. PMID- 15812445 TI - Pediatricians beware: the age of ARTs is upon us. PMID- 15812446 TI - Patterns of brain injury in term neonatal encephalopathy. AB - OBJECTIVES: To determine whether the pattern of brain injury in term neonatal encephalopathy is associated with distinct prenatal and perinatal factors and to determine whether the pattern of injury is associated with 30-month neurodevelopmental outcome. STUDY DESIGN: A total of 173 term newborns with neonatal encephalopathy from 2 centers underwent magnetic resonance imaging (MRI) at a median of 6 days of age (range, 1-24 days). Patterns of injury on MRI were defined on the basis of the predominant site of injury: watershed predominant, basal ganglia/thalamus predominant, and normal. RESULTS: The watershed pattern of injury was seen in 78 newborns (45%), the basal ganglia/thalamus pattern was seen in 44 newborns (25%), and normal MRI studies were seen in 51 newborns (30%). Antenatal conditions such as maternal substance use, gestational diabetes, premature rupture of membranes, pre-eclampsia, and intra-uterine growth restriction did not differ across patterns. The basal ganglia/thalamus pattern was associated with more severe neonatal signs, including more intensive resuscitation at birth ( P = .001), more severe encephalopathy ( P = .0001), and more severe seizures ( P = .0001). The basal ganglia/thalamus pattern was associated with the most impaired motor and cognitive outcome at 30 months. CONCLUSION: The patterns of brain injury in term neonatal encephalopathy are associated with different clinical presentations and neurodevelopmental outcomes. Measured prenatal risk factors did not predict the pattern of brain injury. PMID- 15812447 TI - Growth and development of preterm infants fed infant formulas containing docosahexaenoic acid and arachidonic acid. AB - OBJECTIVES: To evaluate safety and benefits of feeding preterm infants formulas containing docosahexaenoic acid (DHA) and arachidonic acid (ARA) until 92 weeks postmenstrual age (PMA), with follow-up to 118 weeks PMA. STUDY DESIGN: This double-blinded study of 361 preterm infants randomized across three formula groups: (1) control, no supplementation; (2) algal-DHA (DHA from algal oil, ARA from fungal oil); and (3) fish-DHA (DHA from fish oil, ARA from fungal oil). Term infants breast-fed > or =4 months (n = 105) were a reference group. Outcomes included growth, tolerance, adverse events, and Bayley development scores. RESULTS: Weight of the algal-DHA group was significantly greater than the control group from 66 to 118 weeks PMA and the fish-DHA group at 118 weeks PMA but did not differ from term infants at 118 weeks PMA. The algal-DHA group was significantly longer than the control group at 48, 79, and 92 weeks PMA and the fish-DHA group at 57, 79, and 92 weeks PMA but did not differ from term infants from 79 to 118 weeks PMA. Supplemented groups had higher Bayley mental and psychomotor development scores at 118 weeks PMA than did the control group. Supplementation did not increase morbidity or adverse events. CONCLUSIONS: Feeding formulas with DHA and ARA from algal and fungal oils resulted in enhanced growth. Both supplemented formulas provided better developmental outcomes than unsupplemented formulas. PMID- 15812448 TI - Trends in severe bronchopulmonary dysplasia rates between 1994 and 2002. AB - OBJECTIVE: To examine temporal trends in the rates of severe bronchopulmonary dysplasia (BPD) between 1994 and 2002. STUDY DESIGN: In a retrospective cohort study, all infants with a gestational age (GA) <33 weeks in a large managed care organization were identified. Annual rates of BPD (defined as an oxygen requirement at 36 weeks corrected GA), severe BPD (defined as respiratory support at 36 weeks corrected GA), and death before 36 weeks corrected GA were examined. RESULTS: Of the 5115 infants in the study cohort, 603 (12%) had BPD, including 246 (4.9%) who had severe BPD. There were 481 (9.5%) deaths before 36 weeks corrected GA. Although the decline in BPD in this period was not significant, the rates of severe BPD declined from 9.7% in 1994 to 3.7% in 2002. Controlling for gestational age, the odds ratio (95% CI) for annual rate of decline in severe BPD was 0.890 (0.841-0.941). Controlling for gestational age, deaths before 36 weeks corrected GA also declined, with the odds ratio (CI) for the annual decline being 0.944 (0.896-0.996). CONCLUSIONS: In this study population, the odds of having of BPD remained constant after controlling for GA. However, the odds of having severe BPD declined on average 11% per year between 1994 and 2002. PMID- 15812449 TI - Racial and etiopathologic dichotomies in insulin hypersecretion and resistance in obese children. AB - OBJECTIVES: To assess insulin dynamics to oral glucose tolerance testing in obese children, denoting individual contributions of insulin hypersecretion versus resistance to racial and etiopathogenetic specificity. STUDY DESIGN: We performed 3-hour oral glucose tolerance testing in 113 nondiabetic obese children (age 13.6 +/- 3.1 years; 41 male, 78 female; 37 black, 41 white; 35 with central nervous system [CNS] insult). The corrected insulin response (CIRgp; measuring beta-cell secretion) and the composite insulin sensitivity index (CISI) were computed and log-transformed, and each was modeled in terms of the other, plus race/etiology, age, sex, body mass index z score, glucose tolerance, pubertal status, and geographic location. RESULTS: A scatterplot of logCIRgp versus logCISI showed that racial and etiopathogenetic groups plotted in different areas. CISI (controlled for CIRgp and other variables) was only 13% lower in blacks than in whites ( P = .32). Conversely, CIRgp (controlled for CISI and other variables) was 49% higher in blacks ( P = .028). CNS insult exhibited a 40% higher CIRgp ( P = .054) and 11% higher CISI ( P = .42) than intact white subjects. CONCLUSIONS: Insulin hypersecretion and resistance are distinct phenomena in childhood obesity. Insulin hypersecretion appears to be the more relevant insulin abnormality both in obese blacks and in CNS insult. PMID- 15812450 TI - A population-based comparison of BMI percentiles and waist-to-height ratio for identifying cardiovascular risk in youth. AB - OBJECTIVE: Determine whether waist-to-height ratio (WHtR) or sex- and age specific percentiles of body mass index (BMI) better identifies cardiovascular risk. STUDY DESIGN: The third National Health and Nutrition Examination Survey (NHANES III) provided measurements on 7657 participants statistically weighted to represent 50.05 million youth 4 to 17 years of age. We estimated the subpopulations corresponding to BMI strata that were normal (< 85th percentile), at risk for overweight (85th to <95th percentile), and overweight (> or =95th percentile). We chose WHtR cutoff points (0.490 and 0.539) so that subpopulation sizes in the three WHtR strata would equal those in the three BMI strata. For 13 cardiovascular risk factors we compared mean levels among youth discordant for their BMI and WHtR strata. RESULTS: 726 participants (representing 3.69 million youth) were identified as having WHtR stratum >BMI stratum. Compared with the 603 participants (representing 3.70 million youth) who were discordant in the opposite direction, weighted analyses showed they had higher mean levels of heart rate, low-density lipoprotein (LDL) cholesterol, fasting triglycerides, and total cholesterol ( P <.015, adjusted for sex, age, and race-ethnicity). Their mean systolic blood pressure was lower, but this difference was eliminated after adjustment for their shorter stature. CONCLUSION: WHtR, a simpler anthropometric index than sex- and age-specific BMI percentiles, better identifies youth with adverse cardiovascular risk factors. PMID- 15812451 TI - Combining unprotected pancreatic enzymes with pH-sensitive enteric-coated microspheres does not improve nutrient digestion in patients with cystic fibrosis. AB - OBJECTIVES: To assess the efficacy of combining unprotected powder enzymes and oral enteric-coated microsphere (ECM) and to ECM alone in treating nutrient maldigestion in patients with cystic fibrosis. STUDY DESIGN: Patients were randomly assigned into 2 consecutive, 2-week phases; ECM alone, and ECM plus unprotected powder enzymes. Fecal fat, energy, and nitrogen output were compared with intake at the end of each phase. Two-tailed, paired t tests were performed to compare outcomes. RESULTS: The mean age of the 14 patients (3 girls) was 5.7 +/- 3.2 years (range, 1.9 to 13.4 years). There was no significant difference in percent malabsorption of fat (15.6% vs 18.2%), energy (13.3% vs 13.4%), or nitrogen (11.8% vs 11.3%) between phases. CONCLUSIONS: The addition of powder enzymes to ECM did not improve nutrient maldigestion compared with ECM alone. PMID- 15812452 TI - Need for quantitative assessment of transglutaminase autoantibodies for celiac disease in screening-identified children. AB - OBJECTIVES: To assess several transglutaminase autoantibody (TGAA) assays in their ability to distinguish celiac disease (CD) in screening-identified children with abnormal intestine biopsy specimens from those with normal biopsy specimens. STUDY DESIGN: Children at risk for CD (n = 54) composed of type 1 diabetics, first-degree relatives of type 1 diabetics or CD, and HLA-DQ2+ individuals followed from birth received intestine biopsy. Sera obtained at the time of biopsy were tested for TGAA, using the radioimmunoassay and 5 other commercially available enzyme-linked immunosorbent assays. RESULTS: False-positive rates ranged from 28% to 80%. The positive predictive value (PPV) of the tests ranged from 63% to 84% (lower than reported for symptomatic children). Setting a higher cutoff for each assay maximized PPV. CONCLUSIONS: There are significant quantitative differences among all TGAA assays that could affect interpretation of a positive test for CD. The overall false-positive rate for all assays was high in this population. Using the assay as a quantitative rather than qualitative tool by increasing the cutoff of positivity to indicate biopsy increases PPV. Multicenter workshops are needed to identify critical differences and to standardize TGAA assays among laboratories. PMID- 15812453 TI - Gastric sensory and motor dysfunction in adolescents with functional dyspepsia. AB - OBJECTIVES: Validated, noninvasive studies were used to compare sensation and motor function of the upper gastrointestinal tract in adolescents with functional dyspepsia (FD) and in control subjects. STUDY DESIGN: Fifteen adolescents with FD and 15 healthy participants underwent standardized symptom assessment, a satiation nutrient drink test, and 13 C-Spirulina platensis breath test for gastric emptying of solids. Adolescents with FD also underwent measurements of fasting and postprandial gastric volume by means of single-photon emission computed tomography, and their results were compared with those from 15 healthy volunteers (age, 18 to 25 years). RESULTS: Compared with control subjects, adolescents with FD had significantly higher postprandial symptoms 30 minutes after reaching maximum satiation with the nutrient drink test and significant delay in the T 1/2 for gastric emptying of solids. Compared with healthy 18- to 25-year-old adults, adolescents had a diminished postprandial gastric volume response. By means of single-photon emission computed tomography, frequent baseline dyspeptic symptoms were associated with prolonged T 1/2 for gastric emptying and higher postprandial aggregate symptom score. A baseline increased severity of dyspepsia symptoms was associated with prolonged T 1/2 for gastric emptying. CONCLUSIONS: Adolescents with FD demonstrate increased postprandial symptoms after challenge, delayed gastric emptying, and a reduced gastric volume response to feeding. PMID- 15812454 TI - Impact of zinc supplementation on mental and psychomotor scores of children aged 12 to 18 months: a randomized, double-blind trial. AB - OBJECTIVE: To evaluate the effect of zinc supplementation on mental and psychomotor scores in children aged 12 to 18 months. STUDY DESIGN: In this double blind, randomized, placebo-controlled trial, children aged 6 to 30 months received daily elemental zinc (10 mg for infants and 20 mg for others) or placebo for 4 months. Bayley Scales of Infant Development II were used for development assessment in the 12- to 18-month subgroup at enrollment and the end of the study. RESULTS: At the end of the study, the adjusted mean mental ( P = .36) and psychomotor ( P = .28) index scores were similar in the intervention and control groups. In a multivariate model, the baseline mental development index score was positively associated with the mother's schooling, the child's height for age, packed cell volumes, hospital birth, and attendance at a day care center, and was negatively associated with the child's age. Breastfeeding, the child's weight for height, and packed cell volumes were positively associated with the baseline psychomotor index score. CONCLUSION: Zinc supplementation did not affect the mental or psychomotor development index scores in a setting in which zinc deficiency is common. PMID- 15812455 TI - Social consequences in adult life of end-stage renal disease in childhood. AB - OBJECTIVE: To describe employment achievement and social independence of adults with childhood end-stage renal disease (ESRD) and to explore determining factors. STUDY DESIGN: Employment, occupational level, living arrangements, social engagements, and subjective health perception were cross-sectionally established between 1998 and 2000 in 144 of all living 187 adult Dutch patients with ESRD with an onset at age 0 to 15 years between 1972 and 1992. Potential clinical determinants were established by means of a review of all medical charts. RESULTS: Compared with age-matched Dutch citizens, patients were more often involuntarily unemployed (19.4% vs 11.1%), had a lower occupational level, more often still lived with their parents, and more often had no partner. A low occupational level was associated with a dialysis duration >8 years (OR, 9.6; 95% CI, 1.9-47.6); living at the parental home was associated with the male sex (OR, 3.4; 95% CI, 1.5-7.8) and with a dialysis duration >8 years (OR, 3.7; 95% CI, 1.3 10.2). CONCLUSION: Prolonged dialysis during childhood may decrease the ability to gain high-skilled professions and social independence. Unemployment is twice as high in adult patients with childhood ESRD than in healthy persons, but more than twice as low as compared with young ESRD patients with an adult onset of the disease, according to previous reports. PMID- 15812456 TI - Impact of sexual networks on risk for gonorrhea and chlamydia among low-income urban African American adolescents. AB - OBJECTIVE: To determine whether African American adolescents, whose recent sex partners reported having another sex partner, are at increased risk for exposure to genital chlamydial infection or gonorrhea. STUDY DESIGN: A household sample of low-income urban African American adolescents 14 to 19 years of age, up to two of their close friends, and their sex partners were interviewed and tested for gonorrhea and chlamydial infection. RESULTS: Thirty-four of 145 adolescents had at least one recent sex partner infected with Neisseria gonorrhoeae and/or Chlamydia trachomatis. The adjusted model showed that adolescents, whose recent sex partners reported having another sex partner, were more likely to have a recent sex partner with gonorrhea and/or chlamydial infection. CONCLUSION: In addition to individual factors, network factors may explain why African American adolescents are at increased risk for exposure to sexually transmitted infections (STIs). Multi-level community-based interventions may need to address network factors along with personal behaviors in order to prevent STIs among low-income urban African American adolescents. PMID- 15812457 TI - Outbreak of adenovirus type 30 in a neonatal intensive care unit. AB - OBJECTIVES: To describe an outbreak of adenovirus, type 30, in a neonatal intensive care unit (NICU). STUDY DESIGN: This was a retrospective, observational study. RESULTS: An outbreak of adenovirus infection occurred in an NICU. It lasted 6 months and involved 21 of 333 (6.3%) infants. The introduction of infection control measures controlled the outbreak; however, premature discontinuation of the measures resulted in a second wave of cases. The virus caused pneumonia in 7 infants, conjunctivitis in 7 infants, pneumonia and conjunctivitis in 1 infant, and upper respiratory tract illness in 1 infant. Infection was asymptomatic in 5 infants. Six infants died. Death was associated with the presence of pneumonia ( P = .0001), administration of steroids ( P = .003), and mechanical ventilation ( P = .02). Investigation into the origin of the outbreak suggested that the virus may have been introduced and spread during ophthalmologic procedures. CONCLUSIONS: Adenovirus type 30 can cause severe disease among premature infants in an NICU. Infants with severe bronchopulmonary dysplasia requiring mechanical ventilation are more likely to have development of adenovirus pneumonia and die. Standard infection control measures are effective in controlling an outbreak. Ophthalmologic procedures continue to be a potential source of adenovirus outbreaks. PMID- 15812458 TI - Adrenal insufficiency in asymptomatic adrenoleukodystrophy patients identified by very long-chain fatty acid screening. AB - OBJECTIVE: Plasma assay for very long-chain fatty acids has made it possible to perform large-scale screening of at-risk individuals to identify asymptomatic patients with X-linked adrenoleukodystrophy (X-ALD). We evaluated the burden of undiagnosed adrenal insufficiency in 49 such patients (age, 4.5 +/- 3.5 years). STUDY DESIGN: Serum adrenocorticotropic hormone (ACTH) and standard-dose ACTH stimulation test were performed at the baseline and followed prospectively until initiation of adrenal replacement therapy (follow-up, 2 +/- 1.7 years). RESULTS: At baseline, 39 (80%) patients had impaired adrenal function, serum ACTH levels were elevated in 34 (69%) patients, and ACTH stimulation test was abnormal in 21(43%) patients. There was a moderate association between Serum ACTH and age at baseline, ( r = 0.32, P = .05). By the end of follow-up, 86% of patients had borderline or overt adrenal insufficiency (age of onset, 4.8 +/- 3.7 years). CONCLUSIONS: We detected a high prevalence of unrecognized adrenocortical insufficiency in asymptomatic boys with X-ALD. It is known to be a frequent cause of morbidity and can be prevented by careful monitoring, early identification of impaired adrenal reserve, and timely initiation of therapy. It manifests early and before onset of neurologic symptoms, suggesting X-ALD as a candidate disorder for neonatal screening. PMID- 15812459 TI - Hashitoxicosis in children: clinical features and natural history. AB - OBJECTIVE: To determine the incidence, natural history, and clinical characteristics of Hashitoxicosis (Htx) in pediatric patients with autoimmune thyroiditis. STUDY DESIGN: Medical records of children diagnosed with Hashimoto thyroiditis between 1993 and 2002 were reviewed. The clinical course of patients presenting with hyperthyroidism was determined. Variables including sex, age, family history, thyroid hormone levels, anti-thyroid antibody titers, 123 I thyroid scan results, and presenting features were investigated as possible predisposing factors for the development of Htx. RESULTS: Out of 69 patients with autoimmune thyroiditis, 8 were diagnosed with Htx. The duration of hyperthyroidism ranged from 31 to 168 days. Three patients became hypothyroid after an average of 46.3 +/- 13.2 days, and 5 patients became euthyroid after an average of 112.8 +/- 59.8 days. Additional findings included an elevated thyroid stimulating immunoglobulin (TSI) titer in 3 of the 8 patients with Htx, and increased uptake on 123 I scan in 2 patients. CONCLUSION: Htx is an uncommon yet important cause of hyperthyroidism in children that has a variable clinical course. The diagnosis may be complicated, as presenting features sometimes exhibit significant overlap with Graves' disease. No factors predisposing to the development of Htx were identified. PMID- 15812461 TI - Midline defects in FG syndrome: does tethered spinal cord contribute to the phenotype? AB - OBJECTIVES: FG syndrome is an X-linked recessive mental retardation syndrome with ano-rectal anomalies, constipation, and occasional urinary incontinence. Because tethered spinal cord syndrome (TCS) has similar symptoms, we evaluated imaging for TCS in patients with FG syndrome. STUDY DESIGN: Patients were recruited from the International FG Syndrome Support Group, and an FG Syndrome Consensus Group reviewed clinical histories, medical records, and photographs of each responding patient. Results of cranial and spinal imaging studies were available for 12 patients. RESULTS: Of 12 boys with FG syndrome, 6 had hypoplasia of the corpus callosum, and 3 of these had TCS (all with constipation and urinary symptoms). The other 9 did not have urinary symptoms. After surgical untethering, bowel and bladder symptoms improved. CONCLUSIONS: Tethered spinal cord syndrome occurred in 25% of patients with FG syndrome associated with hypoplasia of the corpus callosum and causing bowel and bladder incontinence. A high index of suspicion is necessary for early diagnosis, and timely intervention results in significant improvement in symptomatology. PMID- 15812462 TI - Serum citrulline levels correlate with enteral tolerance and bowel length in infants with short bowel syndrome. AB - OBJECTIVE: To determine if serum levels of CIT (a nonprotein amino acid synthesized by the intestine) correlate with total parenteral nutrition (PN) independence in children with short bowel syndrome (SBS). STUDY DESIGN: We prospectively obtained serum amino acid profiles over a 24-month interval from all infants with SBS 3 weeks to 4 years of age. Remaining small intestine length was recorded at surgery, and percent enteral calories tolerated (enteral calories divided by enteral plus parenteral calories x 100) was determined in 24 infants with SBS and 21 age-matched controls (blood drawn for non-gastrointestinal symptoms). RESULTS: Mean CIT for controls was 31 +/- 2 micromol/L. In patients with SBS (n = 24), serum CIT correlated linearly with percent enteral calories (R = 0.85; P <.001) and with bowel length (R = 0.47; P < or =.03). CIT level in patients with SBS weaned off PN was 30 +/- 2 micromol/L; in those subsequently weaned off PN, 20 +/- 2 micromol/L; and in those who would remain PN-dependent, 11 +/- 2 micromol/L ( P < or =.01). Serum CIT > or =19 micromol/L had 94% sensitivity and 67% specificity for being off or coming off total PN. CONCLUSIONS: Serum CIT level >19 micromol/L in children with SBS is associated with development of enteral tolerance and may be a useful predictive test. PMID- 15812463 TI - Creatine kinase MB (CK-MB) in benign paroxysmal vertigo of childhood: a new diagnostic marker. AB - OBJECTIVES: To evaluate the relation between creatine kinase-MB (CK-MB) and benign paroxysmal vertigo in childhood (BPV). STUDY DESIGN: We prospectively evaluated and followed serum CK-MB in 22 children with BPV diagnosed between 1998 and 2003. RESULTS: The average age of debut for BPV was 1.7 years, and follow-up time was 2.8 years. The CK-MB values were elevated in all children. CK-MB values were persistently increased (mean, 6.0 microg/L) during the study period and were not related to duration of BPV, time since last attack, or frequency of attacks. CK-MB became normal in 7 children who recovered during the study period. After the initial increased CK-MB value, CK, aspartate aminotransferase, and cardiac troponin I (in 16 children) were measured as markers of muscular disease. CK was slightly increased in 7 (31.8%) and aspartate aminotransferase in 14 (63.6%) of the children. Cardiac troponin I was normal in all children. CONCLUSIONS: In this study, serum CK-MB levels were associated with BPV. These findings indicate a possible muscular involvement in BPV. Further studies will be needed to determine if CK-MB is useful as a diagnostic test for BPV. PMID- 15812465 TI - Financial compensation to adolescents for participation in biomedical research: adolescent and parent perspectives in seven studies. AB - OBJECTIVE: To examine the impact of financial compensation on pediatric asthma research participation decision-making and determine whether perceptions of fair compensation differed for parents and adolescents, lower and higher income participants, and compensation-informed and uninformed participants in minimal and above minimal risk research. STUDY DESIGN: Adolescents (n = 36) with asthma and their parents reviewed 7 pediatric asthma research protocols, decided whether they would choose to participate, and provided estimates of "fair" compensation for their participation. Chi-square, analysis of variance, and analysis of covariance were used to determine the affects of compensation on participation and whether various respondents differed in the perceptions of fair compensation. RESULTS: Financial compensation did not affect participation decisions. Estimates of fair compensation were lower for adolescents, lower income respondents, and participants who were naive about potential compensation. Fair compensation estimates were higher than actual compensation for minimal risk studies and lower for above minimal risk studies. CONCLUSIONS: Financial compensation may be a minor consideration in pediatric research participation decision-making. Still, differences in how pediatric researchers and their prospective participants judge fair compensation create the potential for undue influence. Pediatric researchers should use caution when determining a reasonable financial compensation for research participation. PMID- 15812467 TI - The overweight adolescent: clinical and ethical issues in intensive treatments for pediatric obesity. PMID- 15812468 TI - Dizygotic twin pregnancy conceived with assisted reproductive technology associated with chromosomal anomaly, imprinting disorder, and monochorionic placentation. AB - Assisted reproductive technology (ART) has now become a cornerstone of treatment for involuntary infertility. Recent studies have raised concern regarding potential associations between ART and chromosomal aneuploidy, imprinting anomalies, and monochorionic placentation in dizygotic (DZ) twins. We report a case of DZ twins conceived by ART in which all three problems coexist. PMID- 15812469 TI - Cholestasis in children with portal vein obstruction. AB - We describe cholestasis as a result of bile duct abnormalities in 8 children with portal vein obstruction. In a clinical, biochemical and radiological investigation of 121 children with cavernous transformation of the portal vein seen between 1986 and 2000, 8 presented with jaundice, pruritus, and/or raised serum aminotransferases and/or gamma glutamyl transpeptidase (gamma GT) activities. Each displayed dilation and narrowing of intra- and/or extrahepatic bile ducts. Surgical decompression of the portal system (portal-systemic or Rex anastomosis) resulted in the regression of the signs of cholestasis in all children. We conclude that children with portal vein obstruction may exhibit clinically significant cholestasis as a result of external compression of the bile duct by the cavernoma. PMID- 15812470 TI - Orbital varices diagnosed as episcleritis in a child with juvenile idiopathic arthritis. PMID- 15812471 TI - Congenital malalignment of the great toenails mimicking onychomycosis. PMID- 15812472 TI - Treatment of adolescents with gynecomastia. PMID- 15812473 TI - Treatment of adolescents with gynecomastia. PMID- 15812474 TI - "Globesity" and units of measurements. PMID- 15812475 TI - New century: same challenges. PMID- 15812476 TI - A modified Trastuzumab antibody for the immunohistochemical detection of HER-2 overexpression in breast cancer. AB - The immunohistochemical determination of HER-2 to identify patients with advanced breast cancer candidates for Trastuzumab treatment proved neither accurate nor fully reliable, possibly because none of the current reagents detects the specific antigenic site target of Trastuzumab. To circumvent this problem, we conjugated the NH2 groups of Trastuzumab with biotin, and the compound obtained, designated BiotHER, was added directly to tissue sections. Biotin-labelling was revealed with horseradish peroxidase-conjugated streptavidin. Specificity and sensitivity of BiotHER immunostaining with respect to HER-2 amplification were tested on 164 breast carcinoma samples. BiotHER staining was detected on the tumour cell membrane of 12% of all specimens and in 49% specimens with gene amplification, while absent in nonamplified tumours. Predictivity of BiotHER status with respect to the clinical outcome was analysed in 54 patients with HER 2 amplified advanced breast cancer treated with Trastuzumab plus chemotherapy. BiotHER staining, detected in 50% of tumours with HER-2 amplification, was an independent predictor of clinical outcome. In fact, BiotHER positivity was independently associated with increased likelihood of tumour response and reduced risk of tumour progression and death. Biotinylated Trastuzumab can thus be used for immunohistochemical detection of HER-2 overexpression in breast cancer, and has the potential to identify patients likely to benefit from Trastuzumab treatment. PMID- 15812477 TI - Cancer following hip and knee arthroplasty: record linkage study. AB - Concerns have been raised that degradation of implants used in hip and knee arthroplasty may lead to an increased risk of some cancers, particularly those of the haematopoietic, lymphatic and urinary systems. We used linked statistical records of hospital admissions and deaths to compare cancer rates in cohorts of people who had undergone hip or knee arthroplasty with a comparison cohort. We did not find an elevated risk for cancer, overall, in either the hip or knee cohort or in both combined (rate ratio for both combined 0.99; 95% confidence intervals 0.95-1.02), or for haematopoietic, lymphatic or urinary system cancers. There was also no elevation in risk of cancer more than 10 years after arthroplasty. Our findings add to the evidence that arthroplasty is safe in respect of cancer risk. PMID- 15812478 TI - A cohort study of antihypertensive treatments and risk of renal cell cancer. AB - We studied 335,682 county residents, of whom 113,298 had been prescribed antihypertensive treatment (AHT), in the period 1989-2002 in North Jutland County, Denmark to examine the relation between different AHTs and the risk of renal cell carcinoma (RCC). An internal comparison was performed among the different classes of AHT users with users of beta blockers as the reference, in order to address potential confounding and bias. The average follow-up was 10 years (range 0-13). Use of any AHT was associated with RCC (relative rate (RR)=1.6, 95% confidence interval (CI) 1.3-1.9) compared with nonusers in the general population. Specific classes of AHTs were nonsignificantly associated with RCC, but compared with users of beta blockers, the numbers observed were close to expectation. Analyses by duration of follow-up and number of prescriptions revealed no clear trends for any antihypertensive agent and after 5 years of follow-up, the RRs for all classes of AHT decreased. The elevated RRs for RCC among users of AHTs compared with the general population are unlikely to be causal, but rather reflect confounding due to failure to control for pre existing hypertension, and protopathic bias, due to the presence of hypertension as an early sign of kidney disease. PMID- 15812479 TI - Meat consumption and K-ras mutations in sporadic colon and rectal cancer in The Netherlands Cohort Study. AB - Case-cohort analyses were performed on meat and fish consumption in relation to K ras mutations in 448 colon and 160 rectal cancers that occurred during 7.3 years of follow-up, excluding the first 2.3 years, and 2948 subcohort members of The Netherlands Cohort Study on diet and cancer. Adjusted incidence rate ratios and 95% confidence intervals were computed for colon and rectal cancer and for K-ras mutation status subgroups. Total fresh meat, most types of fresh meat and fish were not associated with colon or rectal cancer, neither overall nor with K-ras mutation status. However, several weak associations were observed for tumours with a wild-type K-ras, including beef and colon tumours, and an inverse association for pork with colon and rectal tumours; for meat products, an increased association was observed with wild-type K-ras tumours in the colon and possibly with G>A transitions in rectal tumours. PMID- 15812482 TI - Introducing new vaccines into developing countries: obstacles, opportunities and complexities. AB - Infectious diseases are thought to account for nearly 25% of all deaths worldwide, and extract a disproportionate toll in developing countries. Moreover, infectious diseases are now appreciated to be major causes of the poverty and economic underdevelopment that characterize the world's poorest countries. Development and deployment of new vaccines to prevent infectious diseases in developing countries have therefore become high priorities in the global health agenda. PMID- 15812483 TI - The process development challenge for a new vaccine. AB - The challenges of vaccine development are not limited to identification of suitable antigens, adjuvants and delivery methods, but include regulatory, technical and manufacturing hurdles in translating a vaccine candidate to the clinic. Process development is the technological foundation that underlies the manufacture of new vaccines and is central to successful commercialization. PMID- 15812484 TI - Maurice Hilleman. PMID- 15812485 TI - Vaccines in the public eye. AB - Preventive vaccines are widely acknowledged as the best hope for protection against infectious pathogens such as avian flu, HIV and SARS. As a result, they have received much recent attention in the media that has exposed some of the challenges involved in optimally using vaccine technology. PMID- 15812486 TI - T cell vaccines for microbial infections. AB - Vaccination, or the deliberate induction of protective immunity by administering nonpathogenic forms of a microbe or its antigens to induce a memory immune response, is the world's most cost-effective medical procedure for preventing morbidity and mortality caused by infectious disease. Historically, most vaccines have worked by eliciting long-lived plasma cells. These cells produce antibodies that limit disease by neutralizing a toxin or blocking the spread of the infectious agent. For these 'B cell vaccines,' the immunological marker, or correlate, for protection is the titer of protective antibodies. With the discovery of HIV/AIDS, vaccine development has been confronted by an agent that is not easily blocked by antibody. To overcome this, researchers who are developing HIV/AIDS vaccines have turned to the elicitation of cellular immunity, or 'T cell vaccines,' which recognize and kill infected cells. PMID- 15812487 TI - A real shot. PMID- 15812488 TI - Annulling a dangerous liaison: vaccination strategies against AIDS and tuberculosis. AB - Human immunodeficiency virus (HIV) and Mycobacterium tuberculosis annually cause 3 million and 2 million deaths, respectively. Last year, 600,000 individuals, doubly infected with HIV and M. tuberculosis, died. Since World War I, approximately 150 million people have succumbed to these two infections--more total deaths than in all wars in the last 2,000 years. Although the perceived threats of new infections such as SARS, new variant Creutzfeldt-Jakob disease and anthrax are real, these outbreaks have caused less than 1,000 deaths globally, a death toll AIDS and tuberculosis exact every 2 h. In 2003, 40 million people were infected with HIV, 2 billion with M. tuberculosis, and 15 million with both. Last year, 5 million and 50 million were newly infected with HIV or M. tuberculosis, respectively, with 2 million new double infections. Better control measures are urgently needed. PMID- 15812489 TI - Mucosal immunity and vaccines. AB - There is currently great interest in developing mucosal vaccines against a variety of microbial pathogens. Mucosally induced tolerance also seems to be a promising form of immunomodulation for treating certain autoimmune diseases and allergies. Here we review the properties of the mucosal immune system and discuss advances in the development of mucosal vaccines for protection against infections and for treatment of various inflammatory disorders. PMID- 15812490 TI - Vaccines: past, present and future. AB - The vaccines developed over the first two hundred years since Jenner's lifetime have accomplished striking reductions of infection and disease wherever applied. Pasteur's early approaches to vaccine development, attenuation and inactivation, are even now the two poles of vaccine technology. Today, purification of microbial elements, genetic engineering and improved knowledge of immune protection allow direct creation of attenuated mutants, expression of vaccine proteins in live vectors, purification and even synthesis of microbial antigens, and induction of a variety of immune responses through manipulation of DNA, RNA, proteins and polysaccharides. Both noninfectious and infectious diseases are now within the realm of vaccinology. The profusion of new vaccines enables new populations to be targeted for vaccination, and requires the development of routes of administration additional to injection. With all this come new problems in the production, regulation and distribution of vaccines. PMID- 15812491 TI - Can successful vaccines teach us how to induce efficient protective immune responses? AB - Some recently introduced vaccines that have excellent efficacy records have been developed without a clear understanding of their mechanism of protection. In fact, successful vaccines have often emerged out of empirical observations and have only rarely been the result of a rational use of the continuously increasing immunological knowledge available to scientists. However, a posteriori deciphering of the biological bases for the efficacy of successful vaccines should be an essential component of research efforts directed at the development of new vaccines for the most challenging infectious diseases. PMID- 15812492 TI - Targeting the innate immune response with improved vaccine adjuvants. AB - Despite two centuries of vaccine use, only a few adjuvants and delivery systems are licensed for human use. This is partly because traditional vaccines based on attenuated live organisms already have them--their invasiveness provides efficient delivery to antigen-presenting cells and various naturally occurring components of the pathogens stimulate the innate immune system. But consideration of these immune potentiators and delivery systems has become important to the development of new subunit vaccines consisting of isolated antigens. Here we consider rational approaches to the discovery and development of immunostimulatory compounds and vaccine formulations that target innate immune responses. PMID- 15812493 TI - Peptide-based therapeutic vaccines for allergic and autoimmune diseases. AB - Allergic and autoimmune diseases are forms of immune hypersensitivity that increasingly cause chronic ill health. Most current therapies treat symptoms rather than addressing underlying immunological mechanisms. The ability to modify antigen-specific pathogenic responses by therapeutic vaccination offers the prospect of targeted therapy resulting in long-term clinical improvement without nonspecific immune suppression. Examples of specific immune modulation can be found in nature and in established forms of immune desensitization. Understanding and exploiting common mechanisms such as the ability to induce antigen-specific regulatory cells should allow the development of effective therapeutic strategies for both forms of immunopathology. Targeting pathogenic T cells using vaccines consisting of synthetic peptides representing T cell epitopes is one such strategy that is currently being evaluated with encouraging results. Future challenges in the development of therapeutic vaccines include selection of appropriate antigens and peptides, optimization of peptide dose and route of administration and identifying strategies to induce bystander suppression. PMID- 15812494 TI - Days of Molecular Medicine 2005. PMID- 15812495 TI - Therapies come closer with 'cleaner' stem cell lines. PMID- 15812496 TI - Bitter criticism sours new diabetes research plan. PMID- 15812497 TI - Politics roils US infectious disease center. PMID- 15812498 TI - Money, manpower missing from Europe's new agency. PMID- 15812499 TI - Experts inject reality into cervical cancer prevention schemes. PMID- 15812501 TI - Aggressive HIV strain sets off dubious public health measure. PMID- 15812500 TI - Tysabri withdrawal calls entire class into question. PMID- 15812502 TI - Cancer vaccine field gets shot of optimism from positive results. PMID- 15812505 TI - One step at a time. PMID- 15812506 TI - Statins and control of MHC2TA gene transcription. PMID- 15812509 TI - Can the Institute of Medicine review the FDA? PMID- 15812508 TI - Paracrine action accounts for marked protection of ischemic heart by Akt-modified mesenchymal stem cells. PMID- 15812511 TI - Setting a trap for tissue fibrosis. PMID- 15812513 TI - The shear stress of keeping arteries clear. PMID- 15812514 TI - MET orchestrates cancer and blood coagulation. PMID- 15812515 TI - Pain killer without a high. PMID- 15812516 TI - Beyond beta blockers. PMID- 15812517 TI - Nipping cat allergy with fusion proteins. PMID- 15812518 TI - Getting to the guts of NOD2. PMID- 15812519 TI - HIV unbound. PMID- 15812522 TI - Stem cell transplantation for aplastic anemia and myelodysplastic syndrome. AB - SUMMARY: Stem cell transplantation (SCT) from a histocompatible sibling is treatment of choice for severe aplastic anemia. Survival rates have been reported to be as high as 90% for children. Immunosuppressive therapy (IST) is employed in patients who are not candidates for SCT due to donor unavailability. The addition of cyclosporin A to antilymphocyte globulin has improved the response rate to 70 80%, and survival at 5 years among responders is about 90%. In all, 30% of patients treated by IST suffer from relapse, but long-term prognosis does not appear to be affected by this complication. Juvenile myelomonocytic leukemia (JMML) shares both myelodysplastic and myeloproliferative features. Survival (10 year) of patients with JMML without SCT is only 6%. Children with JMML should be transplanted early in the course of their disease. Conditioning regimen composed of three alkylating agents, busulfan, cyclophosphamide and melphalan has been favored by the EWOG-MDS and EBMT-Pediatric WP in the second half of the 1990s. SCT using this conditioning regimen is capable of curing approximately 50% of patients with JMML. More than 70% of patients with refractory cytopenia and more than 50% of children with advanced MDS are cured of by the early performed allogeneic SCT. PMID- 15812523 TI - Hematopoietic transplantation for bone marrow failure syndromes and thalassemia. AB - Several genetic diseases, generally considered as congenital diseases, are characterized by bone marrow failure during early childhood. Hematopoietic stem cell transplantation is the only curative treatment for syndromes involving bone marrow failure and thalassemia. In this slate-of-the-art review, we wish to focus on the results of hematopoietic transplantation in treating some of these diseases, with a special emphasis on congenital bone marrow failure and thalassemia. The results of this procedure have improved over the previous years, mainly when performed by experienced teams. New conditioning regimes based on fludarabine and the use of HLA-identical donors have been related with better survivals. In the previous years, donors other than HLA-identical siblings have been increasingly used in patients not responding to conventional measures, but this approach needs to be evaluated in larger studies. PMID- 15812524 TI - Hematopoietic cell transplantation activity in Europe for inherited metabolic diseases: open issues and future directions. AB - For the past two decades, hematopoietic cell transplantation (HCT) has been used as effective therapy for selected inherited metabolic diseases (IMD). The primary goals of this therapy have been to promote long-term survival with donor-derived engraftment and to optimize quality of life. Careful, multidisciplinary decision making regarding whether to recommend HCT and how to provide optimal peri- and post-HCT care has proven essential to increase the likelihood of a good outcome. Guidelines for HCT and monitoring have recently been provided in this journal. Here we report data on transplant activity for IMD in Europe and briefly discuss future directions. It is imperative that data collection for these procedures becomes as routine as that for patients undergoing HCT for malignancy and that follow-up is performed in a systematic manner. Large clinical trials have never been performed in this transplant field. Fortunately, accreditation procedures and improvements in information technology can now provide a firm foundation for such trials, which are urgently needed. PMID- 15812525 TI - Treatment of refractory autoimmune diseases with autologous stem cell transplantation: focus on juvenile idiopathic arthritis. AB - Autologous stem cell transplantation (ASCT) can be performed in a variety of refractory autoimmune diseases. A retrospective multicenter analysis is presented to evaluate safety and efficacy of ASCT for refractory juvenile idiopathic arthritis. In all, 18 of the 34 patients (53%) with a follow-up of 12 to 60 months achieved a drug-free complete remission. There were three cases (9%) of transplant-related mortality and two cases of disease-related mortality (6%). Infectious complications were seen frequently. We propose adjustments in future protocols to reduce this mortality in this high-risk patient group. PMID- 15812526 TI - Survey on haematopoietic stem cell transplantation for children in Europe. AB - A recent report, prepared in March 2003, regarding the paediatric transplantation activity registered between 1970 and 2002 in the European Bone Marrow Transplantation (EBMT) database showed a decrease in the number of registrations in 2001 and in 2002. In order to validate this observation, the Paediatric Diseases Working Party (PDsWP) secretariat distributed a questionnaire to 395 institutions participating in the EBMT Registry. Each institution was requested to check the number of transplants they reported and to confirm or to correct the figures. As of 15 March 2004, replies had been received from 135 centres reporting a median of 48 transplants per centre over the study period, total 17 891 (58% of the total number). Among them, 55 confirmed their original figures, while 80 corrected the numbers. The overall number of autologous and allogeneic SCTs performed and not reported were 461 and 692, respectively. Most of the teams that corrected their figures stated that their data managers could provide missing data to the EBMT; 260 other teams, each reporting a median of 15 transplants during the study period, total 12 866 (42% of the total number) chose not to reply. A report prepared in March 2004, following the PDsWP survey, showed an increasing number of transplants performed on patients below 18 years of age between 1973 and 2002 and reported to the EBMT Registry (328 autologous and 628 allogeneic) as compared to the 2003 report. This first PDsWP survey, reaching more than 50% of activity in the field, illustrates that the decrease in activity we observed in the 2003 report does not correspond to a decrease in the number of transplants that were actually performed. It demonstrates the compliance of most major paediatric institutions and confirms the important role of cooperation between National Registries and EBMT Registries. PMID- 15812527 TI - Evolving role of myeloablative chemotherapy in the treatment of childhood brain tumours. AB - Primary brain tumours, a heterogeneous group of cancer that constitute the second most common cancer in childhood, were historically treated with neurosurgical resection and radiation therapy. Chemotherapy has proven to be beneficial for some histological types, which has since led to exploration of the role of high dose chemotherapy and haematopoietic stem cell rescue. Patients with high-grade glial tumours, primitive neuroectodermal tumours and high-risk medulloblastoma usually fare poorly. The indicators of bad prognosis are metastatic status, extent of resection and age. Children <3 years at diagnosis carry worse prognosis. Rare cancers such as ependymoblastoma, atypical teratoid rhabdoid tumour and choroid plexus carcinoma have a dismal prognosis regardless of the above-mentioned indicators. The use of myeloablative therapy (MAT) has been investigated to improve the rate of long-term DFS, as well as to reduce and delay in the youngest children the use of the craniospinal irradiation associated with unacceptable late effects. We will overview the literature regarding patients with 'good and uncertain indications' to MAT. Ependymoma and brain stem tumours, for which the available data discourage the use of MAT, are excluded. Finally, we will summarize a single Institution experience (Giannina Gaslini Children's Hospital, Genoa) with MAT in the period 1997-2003. PMID- 15812528 TI - Graft dysfunction and delayed immune reconstitution following haploidentical peripheral blood hematopoietic stem cell transplantation. AB - For many children with life-threatening hematological diseases, hematopoietic stem cell transplantation (HSCT) is the only curative option. In children lacking a matched related or unrelated donor and with the certainty that, left untreated, death will ensue alternative donors must be sought. Haplo-identical peripheral blood stem cell transplantation (PBSCT) from a healthy parent is a feasible alternative. To reduce the risk of fatal graft-versus-host disease (GvHD) as a complication of transplant across major histocompatibility antigens, intense T cell depletion is required. Large numbers of purified, cytokine mobilized peripheral stem cells (the so-called mega-dose concept) are required to compensate for the significantly increased risk of either graft failure or early rejection. In our unit, despite this approach, graft dysfunction has, in a significant group of children, proved problematic and, despite salvage attempts at re-transplantation, usually fatal. In children with hematological malignant disease, our overall relapse-free survival is 41%. However, successful transplant outcome has been associated with considerable delays in immune reconstitution that can be implicated in subsequent viral reactivation. We are investigating new strategies to improve the outcome of haplo-identical PBSCT, which may allow us to offer this form of treatment to more children requiring urgent HSCT. PMID- 15812529 TI - The hope and the reality of reduced intensity transplants in children with malignant diseases. AB - Reduced intensity preparative regimens are increasingly used for conditioning prior to allogeneic stem cell transplantation. As opposed to classical methods of pre-transplant conditioning, patients receive predominantly immunosuppressive therapies that facilitate early engraftment, while cells within the graft itself promote continuation of the engraftment process. Despite early hopes that this form of transplant would be devoid of grade III and IV acute toxicities, there is a substantial amount of short-term morbidity associated with the technique. Although long-term follow-up is not yet available, it is hoped that these regimens will spare young patients many of the late effects (cataracts, growth retardation, endocrine and reproductive problems) that are often associated with classical pre-transplant conditioning regimens. Reliable engraftment and leukemic control have been demonstrated in a large number of both adult and pediatric transplant recipients of these regimens, many of whom were deemed at high risk for standard conditioning because of serious co-morbidities, previous autologous transplantation or multiply relapsed disease. A brief review of the state of the art of this technology as it applies to pediatric transplantation is presented. Preliminary results of a survey of pediatric transplant centers indicate that a variety of protocols are used for a variety of indications. The use of standardized criteria for implementation of reduced intensity preparative regimens, the use of a limited number of regimens, and more extensive data collection will permit the elaboration of prospective comparative studies of this new and exciting modality. PMID- 15812530 TI - Stem cell transplantation for congenital immunodeficiencies using reduced intensity conditioning. AB - The optimal preparation for stem cell transplantation (SCT) in children with congenital immunodeficiencies is currently unknown. In all, 81 children with immunodeficiency underwent 82 SCTs using reduced-intensity conditioning (RIC). The incidence of significant GVHD was low; viral reactivation was prominent with an unexpected increase in EBV reactivation; immune reconstitution was similar between different donor groups and comparable to conventional SCT. Overall, 68/81 (84%) survive with no significant difference between donor types or between severe combined immunodeficiency (SCID) and non-SCID diseases. Findings suggest a significant survival advantage in the unrelated donor setting for RIC compared to conventional SCT. PMID- 15812531 TI - Is there a role for minimal residual disease levels in the treatment of ALL patients who receive allogeneic stem cells? AB - Relapse is the major complication after allogeneic stem cell transplantation (SCT) for acute lymphoblastic leukemia (ALL) in children. Since it has been possible to measure minimal residual disease (MRD) by real-time quantitative polymerase chain reaction, this parameter is used more frequently in the treatment of ALL. In this article, the role of MRD and chimerism in the treatment and monitoring of pediatric transplantation recipients is described. Pre-SCT MRD levels can predict the risk of relapse and can thus be used to adjust treatment. Post-SCT MRD levels and changes in chimerism can predict relapses as well, although not many treatment options are available today, except relying on a graft-versus-leukemia effect mediated by graft-versus-host disease. Finding new treatments will be the challenge for the near future. PMID- 15812532 TI - Immune reconstitution following hematopoietic progenitor cell transplantation: challenges for the future. AB - Successful hematopoietic progenitor cell transplantation requires rapid and complete transfer of the donor hematopoietic and immune systems to the host. Whereas the uncontrolled transfer of a nontolerant donor immune system results in GVHD in many cases, strategies which diminish GVHD also diminish immune reconstitution. Thus, the reliable, rapid and safe transfer of immunity from donor to host remains a major challenge for the field. Advances in the understanding of the biology of immune reconstitution have elucidated that thymic dependent immune reconstitution can restore global immunity, but is especially vulnerable to toxicities associated with transplant. Alternatively, homeostatic peripheral expansion can be exploited for targeted immunity toward pathogens and tumors, but is difficult to manipulate without exacerbating GVHD risk. New translatable strategies are needed to safely augment one or both of these pathways in the setting of allogeneic hematopoietic progenitor cell transplantation. PMID- 15812533 TI - Immunotherapy in allogeneic hematopoietic stem cell transplantation--not just a case for effector cells. AB - The concept that in allogeneic hematopoietic stem cell transplantation (alloHSCT) the immune system plays a prominent role in the control of leukemic disease is supported by the clinical observation that immunological effector mechanisms contribute to the elimination of leukemic blasts. The failure to induce prolonged remission after alloHSCT has led to resurgent interest in complementing concepts of immune modulation to improve the antileukemic reponse. While the general focus has been placed on manipulation of cytotoxic effector cell populations, we will explore the dual role of leukemia cells as both antigen-presenting and target cells and describe various vaccination strategies to facilitate a protective antileukemic immune response in this setting. In addition, we will introduce mesenchymal stem cells (MSC) as another cell population recently recognized for their immunomodulatory properties. The potential benefits and hazards of MSC cotransplantation in alloHSCT with regard to the graft versus leukemia (GvL) and the graft versus host (GvH) response will be discussed. PMID- 15812534 TI - Novel approaches in GVHD therapy. AB - Severe graft-versus-host disease is a lethal complication to allogeneic haemopoietic stem cell transplantation. This short review gives an overview of novel treatment strategies. Psoralen-enhanced UVA irradiation (PUVA), extracorpoal PUVA, antibodies against IL-2 and TNF-alpha, thalidomide, octreotide, and mesenchymal stem cells are briefly discussed. PMID- 15812535 TI - Update on extracorporeal photochemotherapy for graft-versus-host disease treatment. AB - Pediatric experience with extracorporeal photochemotherapy (ECP) for graft-versus host disease (GvHD) has mainly been reported by Italian and French groups. Data concerning 41 children with acute GvHD and 63 children affected by chronic GvHD are available. In 73 and 63% of them, respectively, improvement was observed, with addition of ECP to their immunosuppressive regimen. Treatment with ECP was associated with minimal side effects, even in the smallest of patients. In all responded pediatric patients, both with acute and chronic GvHD, ECP allowed progressive reduction or discontinuation of the concomitant pharmacological immunosuppressive therapy without an increase in GvHD activity. These data show that ECP is a useful therapy for children affected by GvHD resistant to conventional treatment and can be safely used. PMID- 15812536 TI - Adenovirus infection in children after allogeneic stem cell transplantation: diagnosis, treatment and immunity. AB - Human adenoviruses (HAdV) are a frequent cause of potentially fatal infections in patients after allogeneic stem cell transplantation, especially in children. Monitoring of serum/plasma by real-time quantitative PCR is a sensitive tool for the recognition of patients at risk of a potentially fatal infection and for the evaluation of the efficacy of treatment. Data from a retrospective study and from a prospective study demonstrate that recovery of immunity after transplantation is essential for the elimination of HAdV infection. The feasibility of several approaches for the manipulation of immunity in the immunocompromised host to prevent a fatal course of the infection is discussed. PMID- 15812537 TI - Late effects after stem cell transplantation (SCT) in children--growth and hormones. AB - Stem cell transplantation (SCT) has established itself as a very successful therapy in often otherwise unbeatable disorders. In a subset of children and adolescents there are, however, late effects, often as a combination of the underlying disorder, its primary treatment and subsequent SCT. In children and adolescents, disorders of growth and the endocrine system have been observed to occur frequently. The assurance of normal growth, puberty, fertility and thyroid function--including the prevention of secondary malignancies--is of utmost importance for the overall success of treatment and the maintenance of quality of life. This, however, requires a systematic and structured follow-up programme for patients after SCT. Patients and their families need to be made familiar with this concept early and physicians need to understand that such a system must be implemented as part of a comprehensive care. PMID- 15812538 TI - Secondary malignancies and quality of life after stem cell transplantation. AB - Malignant diseases (MD) occurring after stem cell transplantation (SCT) are of particular concern as increasing number of patients survive and remain free of their original disease. The cumulative incidence at 15 years is 10-12%. The B cell proliferative disorders (BCLP) are the most common MD in the first year after SCT; the incidence probability is 1% in allogeneic transplants but is much higher (until 14%) after HLA-identical, T-cell-depleted SCT in which Campath 1G or ATG are given. BCLP develop because of reactivation of the EBV and a depressed cellular immunity. Prediction of risk of BCLP can be made by frequent monitoring of EBV load in patients with risk factors. The most effective therapies are the early administration of anti-CD20 monoclonal antibody and adoptive immunotherapy with in vitro generated EBV-specific cytotoxic T cells. Myelodysplasia and acute myeloid leukemia with very poor prognosis have been described in 4-18% of patients with non-Hodgkin lymphoma and Hodgkin disease, 12-24 months after autologous SCT. The risk of development of solid tumors increases over time and the cumulative incidence among children who underwent an SCT at less than 10 years of age is 6-11% at 15 years. There are few studies evaluating quality of life (QOL) in children and adolescents who had received an SCT. The findings of these studies can be summarized as follows: (a) The majority of long survivors enjoy good QOL and return successfully to school or work. (b) A minority (10-15%) complain of physical problems or present moderate cognitive or psychological dysfunctions. (c) The importance of family, other social support and psychological adjustments is generally recognized. More extensive, longitudinal and comparative studies with other alternative therapies are required. PMID- 15812540 TI - Allogeneic haematopoietic stem cell transplantation in children with acute lymphoblastic leukaemia: the BFM/IBFM/EBMT concepts. AB - Children with high risk or relapsed acute lymphoblastic leukaemia (ALL) can benefit from allogeneic haematopoietic stem cell transplantation (SCT). To reduce transplantation-associated complications, the BFM study group, the IBFM study group and the PD-WP-EBMT initiated a prospective cooperative multicentre trail for paediatric ALL patients with an indication for allogeneic stem cell transplantation. Four-digit high-resolution HLA typing for all nonsibling donors, standardised GvHD prophylaxis and therapy, uniform conditioning regimen and minimum standards for supportive care should reduce not only treatment-related mortality but also ameliorate late effects for young patients. Furthermore, the prospective evaluation aims to assess the role of haematopoietic SCT in comparison to chemotherapy to enable valuable treatment recommendations for further decisions. PMID- 15812539 TI - The polymorphisms -318C>T in the promoter and 49A>G in exon 1 of CTLA4 and the risk of aplastic anemia in a Caucasian population. AB - Aplastic anemia (AA) is a rare disease with a major autoimmune pathogenetic component. CTLA4 is a T-lymphocyte surface molecule involved in the maintenance of immune tolerance. Some polymorphisms associated with a reduced expression of CTLA4, and thus presumably with increased tendency to autoimmunity, have been associated with various autoimmune diseases. In this study, we evaluated the distribution of the low expression polymorphisms -318C > T and 49A > G of CTLA4 in a population of 67 patients with acquired AA and in 100 normal controls. There was no difference in the distribution of the tested polymorphism between patients and controls and, within the patient group, between those who responded to immunosuppression vs those who did not respond. This study indicates that the polymorphisms -318C > T and 49A > G of CTLA4 do not affect the risk of developing AA and do not influence the response to immunosuppression. PMID- 15812543 TI - Microsatellite instability in patients with chronic B-cell lymphocytic leukaemia. AB - The purpose of our study was to evaluate the microsatellite instability (MSI) at selected loci with known involvement in the oncogenesis of chronic B-cell lymphocytic leukaemia (B-CLL). DNA from B cells (tumour cells) and from T cells (normal controls) of 27 samples of 26 patients with previously untreated B-CLL was extracted. Microsatellite instability in six microsatellite markers was tested using GeneScan Analysis Software. The rate of replication errors positive phenotype (RER+) was determined (MSI in more than 30% of examined loci). RER+ was found in four out of 27 patients (14.8%). A larger proportion of patients with stage C B-CLL exhibited RER+ than those with stage A or B (P < 0.05). A higher prevalence of RER+ was demonstrated in a subgroup of patients with additional malignancies (three out of eight patients) in comparison with patients with B-CLL alone (1/19) (P = 0.031). In conclusion, our study demonstrated that MSI might have a more prominent role in pathogenesis of B-CLL than reported to date. This may result from a selection of microsatellite markers adjacent to chromosomal loci, which are involved in B-cell malignancies, and using GeneScan Analysis Software, which is most modern and precise method of microsatellite analysis. PMID- 15812544 TI - Investigation of systemic folate status, impact of alcohol intake and levels of DNA damage in mononuclear cells of breast cancer patients. AB - Folate is required for DNA synthesis, repair and methylation. Low folate status has been implicated in carcinogenesis, possibly as a result of higher rate of genetic damage. The aim of this study is to compare folate status and levels of DNA damage between breast cancer and benign breast disease control patients. Fasting blood samples from 64 histologically confirmed untreated breast cancer patients (mean age 57 years) and 30 benign breast disease control patients (mean age 51 years) were obtained. Red cell folate (RCF) and plasma homocysteine were measured. Mononuclear cells (MNC) were isolated for genetic damage analysis using the basic alkaline comet assay. Results are expressed as tail moment. Data were log transformed as appropriate before analysis for normalisation purposes. The geometric mean (95% confidence interval) of RCF (ng ml(-1)) in breast cancer patients was 339.07 (333.3-404.6) vs 379.5 (335.8-505.2) in control patients (P = 0.24). Corresponding plasma homocysteine concentrations (micromol l(-1)) were 11.9 (10.6-16.4) vs 10.1 (9.3-11.9) (P = 0.073), respectively. The mean tail moment (s.d.) of DNA damage in MNC of breast cancer patients detected by the basic comet assay was 1.4 (0.66) vs -0.17 (0.79) in controls (P < 0.0001, t test), the modified comet assay 'endonuclease III (Endo III)' was 1.7 (0.70) vs 0.86 (0.81) (P < 0.0001, t-test), and the modified comet assay 'formamidopyrimidine glycosylase (FPG)' was 1.6 (0.62) vs 0.99 (0.94) (P < 0.0001, t-test). There was a significant negative correlation between RCF levels and DNA damage detected by modified comet assay 'FPG' (Pearson Correlation Coefficient r2 = -0.26, P = 0.02) and DNA damage was found to be significantly higher in MNC of breast cancer patients compared to benign breast disease control patients. Breast cancer patients tended to have lower RCF levels and higher levels of plasma homocysteine, but these differences were not significant. The study provides preliminary evidence that reduced folate status may be implicated in the aetiology of breast cancer perhaps by increasing the in vivo level of genetic instability. PMID- 15812545 TI - Induction of effective and antigen-specific antitumour immunity by a liposomal ErbB2/HER2 peptide-based vaccination construct. AB - Efficient delivery of tumour-associated antigens to appropriate cellular compartments of antigen-presenting cells is of prime importance for the induction of potent, cell-mediated antitumour immune responses. We have designed novel multivalent liposomal constructs that co-deliver the p63-71 cytotoxic T Lymphocyte epitope derived from human ErbB2 (HER2), and HA307-319, a T-helper (Th) epitope derived from influenza haemagglutinin. Both peptides were conjugated to the surface of liposomes via a Pam3CSS anchor, a synthetic lipopeptide with potent adjuvant activity. In a murine model system, vaccination with these constructs completely protected BALB/c mice from subsequent s.c. challenge with ErbB2-expressing, but not ErbB2-negative, murine renal carcinoma (Renca) cells, indicating the induction of potent, antigen-specific immune responses. I.v. re challenge of tumour-free animals 2 months after the first tumour cell inoculation did not result in the formation of lung tumour nodules, suggesting that long lasting, systemic immunity had been induced. While still protecting the majority of vaccinated mice, a liposomal construct lacking the Th epitope was less effective than the diepitope construct, also correlating with a lower number of CD8+ IFN-gamma+ T-cells identified upon ex vivo peptide restimulation of splenocytes from vaccinated animals. Importantly, in a therapeutic setting treatment with the liposomal vaccines resulted in cures in the majority of tumour bearing mice and delayed tumour growth in the remaining ones. Our results demonstrate that liposomal constructs which combine Tc and Th peptide antigens and lipopeptide adjuvants can induce efficient, antigen-specific antitumour immunity, and represent promising synthetic delivery systems for the design of specific antitumour vaccines. PMID- 15812546 TI - Validation of a new prognostic index score for disseminated nasopharyngeal carcinoma. AB - Patients with metastatic nasopharyngeal carcinoma have variable survival outcomes. We previously designed a scoring system to better prognosticate these patients. Here, we report results on validation of this new prognostic index score in a separate cohort of patients. Clinical features and laboratory parameters were examined in 172 patients with univariate and multivariate analyses and a numerical score was derived for each independent prognostic variable. Significant independent prognostic variables and their scores assigned included poor performance status (score 5), haemoglobin < 12 g dl(-1) (score 4) and disease-free interval (DFI) (DFI < or = 6 months (score 10) or metastases at initial diagnosis (score 1)). Maximum score was 19 and patients stratified into three prognostic groups: good, 0-3; intermediate, 4-8; poor, > or = 9. When applied to a separate cohort of 120 patients, 59 patients were good, 43 intermediate and 18 poor prognosis, with median survivals of 19.6 (95% CI 16.1, 23.1), 14.3 (95% CI 12.3, 16.2) and 7.9 (95% CI 6.6, 9.2) months, respectively. (logrank test: P = 0.003). We have validated a new prognostic score with factors readily available in the clinics. This simple score will prove useful as a method to prognosticate and stratify patients as well as to promote consistent reporting among clinical trials. PMID- 15812547 TI - HPV type concordance in sexual couples determines the effect of condoms on regression of flat penile lesions. AB - We earlier demonstrated, in a randomised clinical trial, that the regression time of flat penile lesions in male sexual partners of women with cervical intraepithelial neoplasia (CIN) was shorter in men who used condoms compared to those who did not. To further evaluate this finding, we examined whether the effect of condom use on the regression of flat penile lesions depends on the presence of human papillomavirus (HPV) type concordance in sexual couples, as determined in cervical and penile scrapes by GP5+/6+ PCR testing. A Cox model with time-dependent covariates showed a beneficial effect of condoms on regression of flat penile lesions in concordant couples (hazard ratio 2.63, 95% CI 1.07-6.48) but not in those who were nonconcordant. When both partners harboured different HPV types, no effect of condoms was found (hazard ratio 0.90, 95% CI 0.27-2.96). Delayed regression of flat penile lesions was associated with either stable lesions or with new penile lesions developing at sites surrounding pre-existing lesions suggesting reinfection of the penile epithelium. We conclude that condom use blocks sexual HPV transmission by preventing reinfection and development of new penile lesions in men who are susceptible to the same type as present in the female partner. PMID- 15812548 TI - The use of denial in an ethnically diverse British cancer population: a cross sectional study. AB - A total of 82 Asian and 117 randomly selected white Caucasian patients at the Leicestershire Cancer Centre were assessed using measures of coping and adaption to cancer. On the Mental Adjustment to Cancer (MAC) scale, Asian patients were more fatalistic (P < 0.0001) and had more significant hopeless/helpless scores (P = 0.007). The two ethnic groups answered the three questions thought to assess denial differently. Caucasians were more likely not to dwell on their illness (73 vs 55.5%, P < 0.0001) and agree with the statement 'I have difficulty believing this is happening to me' (73 vs 60.5%, P < 0.0001). However, Asian patients were more likely to agree with the statement 'I don't really believe I have cancer' (48.2 vs 31.3%, P = 0.019). Within both groups there was an association with denial and anxious preoccupation (P < 0.001). On the Hospital Anxiety and Depression (HAD) scale, there was no difference in anxiety scores between either sexes or between the Asian and Caucasian groups. However, Asian patients were more depressed (P = 0.001). Although denial was significantly related to the presence of both depression (P < 0.0001) and anxiety (P = 0.001) in the entire patient population, there were different predictors of denial in each subgroup. On multiple regression analysis depression was linked with denial in Caucasians, whereas Fighting Spirit (minus helplessness/hopelessness) was linked with denial in Asian patients. There are definite differences in coping styles in British cancer patients according to ethnicity. While significant numbers in both groups employ denial in some form, Caucasian patients appear to adapt to the psychological pressures of cancer more successfully than Asian patients at a particular point in time. Further work is required to elucidate longitudinal relationships between denial and adaption to cancer. PMID- 15812549 TI - Absence of a specific radiation signature in post-Chernobyl thyroid cancers. AB - Thyroid cancers have been the main medical consequence of the Chernobyl accident. On the basis of their pathological features and of the fact that a large proportion of them demonstrate RET-PTC translocations, these cancers are considered as similar to classical sporadic papillary carcinomas, although molecular alterations differ between both tumours. We analysed gene expression in post-Chernobyl cancers, sporadic papillary carcinomas and compared to autonomous adenomas used as controls. Unsupervised clustering of these data did not distinguish between the cancers, but separates both cancers from adenomas. No gene signature separating sporadic from post-Chernobyl PTC (chPTC) could be found using supervised and unsupervised classification methods although such a signature is demonstrated for cancers and adenomas. Furthermore, we demonstrate that pooled RNA from sporadic and chPTC are as strongly correlated as two independent sporadic PTC pools, one from Europe, one from the US involving patients not exposed to Chernobyl radiations. This result relies on cDNA and Affymetrix microarrays. Thus, platform-specific artifacts are controlled for. Our findings suggest the absence of a radiation fingerprint in the chPTC and support the concept that post-Chernobyl cancer data, for which the cancer-causing event and its date are known, are a unique source of information to study naturally occurring papillary carcinomas. PMID- 15812550 TI - Immunosuppressive effects of radiation on human dendritic cells: reduced IL-12 production on activation and impairment of naive T-cell priming. AB - Dendritic cells (DC) are professional antigen-presenting cells (APC) of the immune system, uniquely able to prime naive T-cell responses. They are the focus of a range of novel strategies for the immunotherapy of cancer, a proportion of which include treating DC with ionising radiation to high dose. The effects of radiation on DC have not, however, been fully characterised. We therefore cultured human myeloid DC from CD14+ precursors, and studied the effects of ionising radiation on their phenotype and function. Dendritic cells were remarkably resistant against radiation-induced apoptosis, showed limited changes in surface phenotype, and mostly maintained their endocytic, phagocytic and migratory capacity. However, irradiated DC were less effective in a mixed lymphocyte reaction, and on maturation produced significantly less IL-12 than unirradiated controls, while IL-10 secretion was maintained. Furthermore, peptide pulsed irradiated mature DC were less effective at naive T-cell priming, stimulating fewer effector cells with lower cytotoxicity against antigen-specific targets. Hence irradiation of DC in vitro, and potentially in vivo, has a significant impact on their function, and may shift the balance between T-cell activation and tolerization in DC-mediated immune responses. PMID- 15812551 TI - The development and characterisation of porphyrin isothiocyanate-monoclonal antibody conjugates for photoimmunotherapy. AB - A promising approach to increase the specificity of photosensitizers used in photodynamic therapy has been through conjugation to monoclonal antibodies (MAb) directed against tumour-associated antigens. Many of the conjugations performed to date have relied on the activated ester method, which can lead to impure conjugate preparations and antibody crosslinking. Here, we report the development of photosensitizer-MAb conjugates utilising two porphyrin isothiocyanates. The presence of a single reactive isothiocyanate allowed facile conjugation to MAb FSP 77 and 17.1A directed against internalizing antigens, and MAb 35A7 that binds to a non-internalizing antigen. The photosensitizer-MAb conjugates substituted with 1-3 mol of photosensitizer were characterised in vitro. No appreciable loss of immunoreactivity was observed and binding specificity was comparable to that of the unconjugated MAb. Substitution with photosensitizer had a minimal effect on antibody biodistribution in vivo for the majority of the conjugates, although a decreased serum half-life was observed using a cationic photosensitizer at the higher loading ratios. Tumour-to-normal tissue ratios as high as 33.5 were observed using MAb 35A7 conjugates. The internalizing conjugate showed a higher level of phototoxicity as compared with the non-internalizing reagent, using a cell line engineered to express both target antigens. These data demonstrate the applicability of the isothiocyanate group for the development of high-quality conjugates, and the use of internalizing MAb to significantly increase the photodynamic efficiency of conjugates during photoimmunotherapy. PMID- 15812552 TI - Indomethacin-induced activation of the death receptor-mediated apoptosis pathway circumvents acquired doxorubicin resistance in SCLC cells. AB - Small-cell lung cancers (SCLCs) initially respond to chemotherapy but are often resistant at recurrence. A potentially new method to overcome resistance is to combine classical chemotherapeutic drugs with apoptosis induction via tumour necrosis factor (TNF) death receptor family members such as Fas. The doxorubicin resistant human SCLC cell line GLC4-Adr and its parental doxorubicin-sensitive line GLC4 were used to analyse the potential of the Fas-mediated apoptotic pathway and the mitochondrial apoptotic pathway to modulate doxorubicin resistance in SCLC. Western blotting showed that all proteins necessary for death inducing signalling complex formation and several inhibitors of apoptosis were expressed in both lines. The proapototic proteins Bid and caspase-8, however, were higher expressed in GLC4-Adr. In addition, GLC4-Adr expressed more Fas (3.1x) at the cell membrane. Both lines were resistant to anti-Fas antibody, but plus the protein synthesis inhibitor cycloheximide anti-Fas antibody induced 40% apoptosis in GLC4-Adr. Indomethacin, which targets the mitochondrial apoptotic pathway, induced apoptosis in GLC4-Adr but not in GLC4 cells. Surprisingly, in GLC4-Adr indomethacin induced caspase-8 and caspase-9 activation as well as Bid cleavage, while both caspase-8 and caspase-9 specific inhibitors blocked indomethacin-induced apoptosis. In GLC4-Adr, doxorubicin plus indomethacin resulted in elevated caspase activity and a 2.7-fold enhanced sensitivity to doxorubicin. In contrast, no effect of indomethacin on doxorubicin sensitivity was observed in GLC4. Our findings show that indomethacin increases the cytotoxic activity of doxorubicin in a doxorubicin-resistant SCLC cell line partly via the death receptor apoptosis pathway, independent of Fas. PMID- 15812553 TI - Gemcitabine, cisplatin and methylprednisolone (GEM-P) is an effective salvage regimen in patients with relapsed and refractory lymphoma. AB - There is currently no standard salvage chemotherapy regimen in relapsed and refractory lymphoma. Gemcitabine is a novel nucleoside analogue, which acts synergistically with cisplatin both in vitro and in clinical studies. We evaluated the combination of gemcitabine, cisplatin and methylprednisolone (GEM P) in 41 heavily pretreated patients with relapsed and refractory Hodgkin's and non-Hodgkin's lymphoma. The best-achieved response rate (RR) was 79% (95% CI 64 91), with a complete RR of 21%. In patients with chemo-resistant disease, the RR was 63%. Myelosuppression was the main toxicity, the incidence of Grade 3 or 4 anaemia, neutropenia and thrombocytopenia was 17.1, 61.0 and 53.7% respectively. Only one patient had neutropenic sepsis and none of the patients suffered from haemorrhage. Grade 3 or 4 nonhaematological toxicity was minimal and stem cell mobilisation was not inhibited. GEM-P is an effective salvage regimen and its use prior to autologous stem cell transplant warrants further investigation. PMID- 15812554 TI - Meta-analysis of randomised adjuvant therapy trials for pancreatic cancer. AB - The aim of this study was to investigate the worldwide evidence of the roles of adjuvant chemoradiation and adjuvant chemotherapy on survival in potentially curative resected pancreatic cancer. Five randomised controlled trials of adjuvant treatment in patients with histologically proven pancreatic ductal adenocarcinoma were identified, of which the four most recent trials provided individual patient data (875 patients). This meta-analysis includes previously unpublished follow-up data on 261 patients. The pooled estimate of the hazard ratio (HR) indicated a 25% significant reduction in the risk of death with chemotherapy (H = 0.75, 95% confidence interval (CI): 0.64, 0.90, P values(stratified) (Pstrat) = 0.001) with median survival estimated at 19.0 (95% CI: 16.4, 21.1) months with chemotherapy and 13.5 (95% CI: 12.2, 15.8) without. The 2- and 5-year survival rates were estimated at 38 and 19%, respectively, with chemotherapy and 28 and 12% without. The pooled estimate of the HR indicated no significant difference in the risk of death with chemoradiation (HR = 1.09, 95% CI: 0.89, 1.32, Pstrat = 0.43) with median survivals estimated at 15.8 (95% CI: 13.9, 18.1) months with chemoradiation and 15.2 (95% CI: 13.1, 18.2) without. The 2- and 5-year survival rates were estimated at 30 and 12%, respectively, with chemoradiation and 34 and 17% without. Subgroup analyses estimated that chemoradiation was more effective and chemotherapy less effective in patients with positive resection margins. These results show that chemotherapy is effective adjuvant treatment in pancreatic cancer but not chemoradiation. Further studies with chemoradiation are warranted in patients with positive resection margins, as chemotherapy appeared relatively ineffective in this patient subgroup. PMID- 15812555 TI - Elucidation of the tumoritropic principle of hypericin. AB - Hypericin is a potent agent in the photodynamic therapy of cancers. To better understand its tumoritropic behaviour, we evaluated the major determinants of the accumulation and dispersion of hypericin in subcutaneously growing mouse tumours. A rapid exponential decay in tumour accumulation of hypericin as a function of tumour weight was observed for each of the six tumour models investigated, and a similar relationship was found between tumour blood flow and tumour weight. Moreover, there was a close correlation between the higher hypericin uptake in RIF-1 tumours compared to R1 tumours and tumour vessel permeability. To define the role of lipoproteins in the transport of hypericin through the interstitial space, we performed a visual and quantitative analysis of the colocalization of hypericin and DiOC18-labelled lipoproteins in microscopic fluorescent overlay images. A coupled dynamic behaviour was found early after injection (normalised fluorescence intensity differences were on the whole less than 10%), while a shifted pattern in localisation of hypericin and DiOC18 was seen after 24 h, suggesting that during its migration through the tumour mass, hypericin is released from the lipoprotein complex. In conclusion, we were able to show that the tumour accumulation of hypericin is critically determined by a combination of biological (blood flow, vessel permeability) and physicochemical elements (affinity for interstitial constituents). PMID- 15812556 TI - Somatostatin receptors 2 and 5 are preferentially expressed in proliferating endothelium. AB - Angiogenesis is characterised by activation, migration and proliferation of endothelial cells and is central to the pathology of cancer, cardiovascular disease and chronic inflammation. Somatostatin is an inhibitory polypeptide that acts through five receptors (sst 1, 2, 3, 4, 5). Sst has previously been reported in endothelium, but their role remains obscure. Here, we report the expression of sst in human umbilical vein endothelial cells (HUVECs) in vitro, during proliferation and quiescence. A protocol for culturing proliferating and quiescent HUVECs was established, and verified by analysing cell cycle distribution in propidium-iodide-stained samples using flow cytometry. Sst mRNA was then quantified in nine proliferating and quiescent HUVEC lines using quantitative reverse transcriptase-polymerase chain reaction. Sst 2 and 5 were preferentially expressed in proliferating HUVECs. All samples were negative for sst 4. Sst 1 and 3 expression and cell cycle progression were unrelated. Immunostaining for sst 2 and 5 showed positivity in proliferating but not quiescent cells, confirming sst 2 and 5 protein expression. Inhibition of proliferating cells with somatostatin analogues Octreotide and SOM230, which have sst 5 activity, was found (Octreotide 10(-10)-10(-6) M: 48.5-70.2% inhibition; SOM230 10(-9)-10(-6) M: 44.9-65.4% inhibition) in a dose-dependent manner, suggesting that sst 5 may have functional activity in proliferation. Dynamic changes in sst 2 and 5 expression during the cell cycle and the inhibition of proliferation with specific analogues suggest that these receptors may have a role in angiogenesis. PMID- 15812557 TI - Histopathologic indicators of breast cancer biology: insights from population mammographic screening. AB - Histopathologic features of breast cancer such as tumour size, grade and axillary lymph node (LN) status variably reflect tumour biology and time. Recent evidence suggests that the biological character of breast cancer is established at an early stage and has a major impact on clinical course. The aim of this study was to distinguish the impact of biology on breast cancer histopathology by comparing features of breast cancers diagnosed following population mammographic screening with prevalent vs incident detection and screening interval. Central histopathology review data from 1147 cases of ductal in situ and/or invasive breast cancer were examined. Size, grade and LN status of invasive cancers were positively correlated (P < 0.001). Prevalent invasive cancers were larger (P < 0.001) and more likely to be LN positive (P = 0.02) than incident cases, but grade was not associated with screening episode (P = 0.7). Screening interval for incident cancers was positively associated with invasive cancer size (P = 0.05) and LN status (P = 0.002) but not grade (P = 0.1). Together, these data indicate that biology and time both impact on size and LN status of invasive breast cancer, but grade reflects biology alone. In view of the clinical importance of breast cancer biology, grade as its most direct indicator assumes particular significance. PMID- 15812558 TI - Characterisation of systemic dissemination of nonreplicating adenoviral vectors from tumours in local gene delivery. AB - Systemic virus dissemination is a potential problem during local gene delivery in solid tumours. However, the kinetics and pathways of the dissemination have not been well characterised during the first 24 h after the infusion is started. To this end, we infused adenoviral vectors for luciferase or enhanced green fluorescence protein into three different tumour models in mice. During and/or after the infusion, we determined the amount of adenoviruses in the tumour, blood, and liver, and examined the transgene expression in the liver, lung, blood, and tumour. In addition, we intravenously injected tumour cells expressing luciferase and examined the biodistribution of these cells in the body. We observed transgene expression in the liver and tumour at 24 h after the infusion, but could not detect transgene expression in the blood and lung. The peak concentration of viral vectors in the plasma occurred during the intratumoral infusion. At 10 min after the infusion, few viral vectors remained in the blood and the ratio of copy numbers of adenoviruses between liver and tumour was > 2 in 80% and > or = 10 in 40% of the mice. Most tumour cells injected intravenously accumulated in the lung within the first 24 h. Taken together, these data indicated that systemic virus dissemination occurred mainly during the first 10 min after the intratumoral infusion was started, and that the dissemination was due to infusion-induced convective transport of viral vectors into leaky tumour microvessels. PMID- 15812559 TI - Vascular endothelial growth factor acts as an osteolytic factor in breast cancer metastases to bone. AB - Vascular endothelial growth factor (VEGF) is a proangiogenic cytokine that is expressed highly in many solid tumours often correlating with a poor prognosis. In this study, we investigated the expression of VEGF and its receptors in bone metastases from primary human breast tumours and further characterised its effects on osteoclasts in vitro. Breast cancer metastases to bone were immunohistochemically stained for VEGF, its receptors VEGFR1 and 2 (vascular endothelial growth factor receptor 1 and 2), demonstrating that breast cancer metastases express VEGF strongly and that surrounding osteoclasts express both VEGFR1 and VEGFR2. RAW 264.7 cells (mouse monocyte cell line) and human peripheral blood mononuclear cells (PBMCs) were cultured with VEGF, RANKL and M CSF. VEGF and RANKL together induced differentiation of multinucleated, tartrate resistant acid phophatase (TRAP)-positive cells in similar numbers to M-CSF and RANKL. The PBMCs were also able to significantly stimulate resorption of mineralised matrix after treatment with M-CSF with RANKL and VEGF with RANKL. We have shown that VEGF in the presence of RANKL supports PBMC differentiation into osteoclast-like cells, able to resorb substrate. Vascular endothelial growth factor may therefore play a role in physiological bone resorption and in pathological situations. Consequently, VEGF signalling may be a therapeutic target for osteoclast inhibition in conditions such as tumour osteolysis. PMID- 15812560 TI - Insulin-like growth factor-I receptor activity is essential for Kaposi's sarcoma growth and survival. AB - Kaposi's sarcoma (KS) is a highly vascular tumour and is the most common neoplasm associated with human immunodeficiency virus (HIV-1) infection. Growth factors, in particular vascular endothelial growth factor (VEGF), have been shown to play an important role in its development. The role of insulin-like growth factors (IGFs) in the pathophysiology of different tumours led us to evaluate the role of IGF system in KS. The IGF-I receptors (IGF-IR) were identified by immunohistochemistry in biopsies taken from patients with different AIDS/HIV related KS stages and on KSIMM cells (an established KS-derived cell line). Insulin-like growth factor-I is a growth factor for KSIMM cells with a maximum increase of 3H-thymidine incorporation of 130 +/- 27.6% (P < 0.05) similar to that induced by VEGF and with which it is additive (281 +/- 13%) (P < 0.05). Moreover, specific blockade of the receptor (either by alpha IR3 antibody or by picropodophyllin, a recently described selective IGF-IR tyrosine phosphorylation inhibitor) induced KSIMM apoptosis, suggesting that IGF-IR agonists (IGF-I and II) mediate antiapoptotic signals for these cells. We were able to identify an autocrine loop essential for KSIMM cell survival in which IGF-II is the IGF-IR agonist secreted by the cells. In conclusion, IGF-I pathway inhibition is a promising therapeutical approach for KS tumours. PMID- 15812561 TI - Twin research: exploring female sexuality. PMID- 15812562 TI - Most DC-SIGNR transcripts at mucosal HIV transmission sites are alternatively spliced isoforms. AB - The repeat region of DC-SIGNR (CD209L) is polymorphic on the genomic level, and, in a separate study, we observed a correlation between the DC-SIGNR genotype and HIV-1 susceptibility during sexual contact. However, previous investigations using immunohistochemistry failed to detect membrane-bound DC-SIGNR on cells in the genital and rectal mucosa. We therefore explored the presence of DC-SIGNR in these compartments with a more sensitive limiting dilution RT-PCR, which also allowed for quantification of alternatively spliced mRNA isoforms. DC-SIGN (CD209) and DC-SIGNR mRNA transcript isoforms were found in all 12 vaginal and two rectal biopsies obtained from 14 healthy individuals. For DC-SIGNR, we detected significantly more isoform than full-length transcripts (mean copy numbers/mug RNA: 602 vs 26; P=0.0009). Four mucosal samples lacked full-length DC SIGNR transcripts entirely. Cloning and sequencing of DC-SIGNR mRNA in three additional individuals revealed a diverse repertoire of DC-SIGNR isoforms, many of which encoded for proteins predicted to be soluble and secreted. Indeed, in one vaginal sample, we detected only soluble isoforms. In conjunction with our prior observation that the DC-SIGNR genotype has an effect on HIV-1 transmission in vivo, these findings emphasize that DC-SIGNR, in addition to DC-SIGN, should be considered as a cofactor in sexual HIV-1 transmission. Soluble isoforms, in particular, may modulate the efficiency of viral transmission and dissemination. PMID- 15812563 TI - Proopiomelanocortin gene variants are associated with serum leptin and body fat in a normal female population. AB - A major quantitative trait locus (QTL) determining leptin levels has been linked to the proopiomelanocortin (POMC) region on chromosome 2. Most studies, based on under 350 lean or obese subjects, have shown no association between POMC SNP 8246 C/T and serum leptin, but significant associations have been reported with RsaI 8246 C/T SNP haplotypes. We have investigated association of four POMC SNPs with body composition and serum leptin in 2758 normal Caucasian female subjects (mean age 47.4+/-12.5 years), from the St Thomas' UK Adult Twin Registry (Twins UK): RsaI and 51 G/C in the 5'UTR and 8246 C/T and 7965 C/T in the 3'UTR. Under the recessive model, the 8246 T allele (freq. 0.18) was significantly associated with higher mean BMI (P=0.032) and total fat (P=0.046, both after age adjustment). Significant associations were maintained in sib-TDT with waist (P=0.049), total fat (P=0.037) and emerged with serum leptin (P=0.016). Initial significant associations between RsaI (-) allele (freq. 0.30) and higher waist (P=0.04) or % central fat (P=0.02) were not maintained in sib-TDT. No significant associations were found between body composition or serum leptin and RsaI/8246 C/T haplotype and none with 51 G/C (freq. 0.01) or 7965 C/T (freq. 0.004). There was minimal pairwise LD between the four loci, apart from RsaI and 8246 C/T (D'=-0.78 (P<0.0001)). Associations of BMI, weight and total fat with SNPs in regions flanking the POMC gene in this powerful study suggest that regulation of POMC expression may be influential in determining body weight. PMID- 15812564 TI - Fine mapping of a schizophrenia susceptibility locus at chromosome 6q23: increased evidence for linkage and reduced linkage interval. AB - We previously reported an autosomal scan for schizophrenia susceptibility loci in a systematically recruited sample of Arab Israeli families. The scan detected significant evidence for linkage at chromosome 6q23 with a nonparametric LOD score (NPL) of 4.60 (P=0.000004) and a multipoint parametric LOD score of 4.16. In order to refine this finding we typed 42 additional microsatellite markers on chromosome 6q between D6S1570 (99.01 cM from the pter) and D6S281 (190.14 from the pter) in the same sample (average intermarker distance approximately 1.7 cM). In the 23 cM region between D6S1715 and D6S311, markers were more closely spaced ( approximately 1.1 cM). Multipoint nonparametric and parametric and single point linkage analyses were performed. The peak NPL rose to 4.98 (P=0.00000058) at D6S1626 (136.97 cM), immediately adjacent to D6S292 (NPL 4.98, P=0.00000068), the marker that gave the highest NPL in the original genome scan, under the broad diagnostic category. The putative susceptibility region (NPL-1) was reduced from 12.0 to 4.96 cM. The peak multipoint parametric LOD score was 4.63 at D6S1626 under a dominant genetic model, core diagnostic category and the LOD-1 interval was 2.10 cM. The maximum single point LOD score (3.55, theta=0.01) was also at D6S1626 (dominant model, core diagnostic category). Increased evidence for linkage in the same sample as in the original genome scan and consistent localization of the linkage peak add further support for the presence of a schizophrenia susceptibility locus at chromosome 6q23. Moreover, the markedly reduced linkage interval greatly improves prospects for identifying a schizophrenia susceptibility gene within the implicated region. PMID- 15812565 TI - A new CARD15 mutation in Blau syndrome. AB - The caspase recruitment domain gene CARD15/NOD2, encoding a cellular receptor involved in an NF-kappaB-mediated pathway of innate immunity, was first identified as a major susceptibility gene for Crohn's disease (CD), and more recently, as responsible for Blau syndrome (BS), a rare autosomal-dominant trait characterized by arthritis, uveitis, skin rash and granulomatous inflammation. While CARD15 variants associated with CD are located within or near the C terminal leucine-rich repeat domain and cause decreased NF-kappaB activation, BS mutations affect the central nucleotide-binding NACHT domain and result in increased NF-kappaB activation. In an Italian family with BS, we detected a novel mutation E383K, whose pathogenicity is strongly supported by cosegregation with the disease in the family and absence in controls, and by the evolutionary conservation and structural role of the affected glutamate close to the Walker B motif of the nucleotide-binding site in the NACHT domain. Interestingly, substitutions at corresponding positions in another NACHT family member cause similar autoinflammatory phenotypes. PMID- 15812566 TI - Cortical NAA deficits in HIV infection without dementia: influence of alcoholism comorbidity. AB - Alcoholism comorbidity is highly prevalent in individuals infected with human immunodeficiency virus (HIV). Each condition is known to affect brain structure, function, and metabolism, but the combined effects on the brain have only recently been considered. Single-voxel, proton MR spectroscopy (MRS) has yielded sensitive measures of early brain deterioration in the progression of HIV, but has limited coverage of neocortex, whereas MRS imaging (MRSI) can simultaneously interrogate large regions of cortex. Included were 15 men with HIV+alcoholism, nine men with HIV alone, eight men with alcoholism alone (abstinent for 3-17 months), and 23 controls. The two HIV groups were matched in T-cell count and were not demented; the two alcoholism groups were relatively matched in lifetime alcohol consumption. We used MRSI with a variable-density spiral sequence to quantify major proton metabolites--N-acetylaspartate (NAA), creatine (Cr), and choline (Cho)-in the superior parietal-occipital cortex. Metabolites were expressed in absolute units and as the NAA/Cr ratio. Significant group effects were present for NAA and Cr. Only the HIV+alcoholism group was significantly affected, exhibiting a 0.8 SD deficit in NAA and a 1.0 SD deficit in Cr. The deficits were not related to highly active antiretroviral therapy (HAART) status. Neither HIV infection nor alcoholism independently resulted in parietal-occipital cortical metabolite abnormalities, yet each disease carried a liability that put affected individuals at a heightened risk of neuronal compromise when the diseases were compounded. Further, the use of absolute measures revealed deficits in NAA and Cr that would have gone undetected if these metabolites were expressed as a ratio. PMID- 15812567 TI - Motivational effects of cannabinoids and opioids on food reinforcement depend on simultaneous activation of cannabinoid and opioid systems. AB - Strong functional interactions exist between endogenous cannabinoid and opioid systems. Here, we investigated whether cannabinoid-opioid interactions modulate motivational effects of food reinforcement. In rats responding for food under a progressive-ratio schedule, the maximal effort (break point) expended to obtain 45 mg pellets depended on the level of food deprivation, with free-feeding reducing break points and food-deprivation increasing break points. Delta-9 tetrahydrocannabinol (THC; 0.3-5.6 mg/kg intrapeitoneally (i.p.)) and morphine (1 10 mg/kg i.p.) dose-dependently increased break points for food reinforcement, while the cannabinoid CB1 receptor antagonist rimonabant (SR-141716A; 0.3-3 mg/kg i.p.) and the preferential mu-opioid receptor antagonist naloxone (0.3-3 mg/kg i.p.) dose-dependently decreased break points. THC and morphine only increased break points when food was delivered during testing, suggesting that these treatments directly influenced reinforcing effects of food, rather than increasing behavior in a nonspecific manner. Effects of THC were blocked by rimonabant and effects of morphine were blocked by naloxone, demonstrating that THC's effects depended on cannabinoid CB1 receptor activation and morphine's effects depended on opioid-receptor activation. Furthermore, THC's effects were blocked by naloxone and morphine's effects were blocked by rimonabant, demonstrating that mu-opioid receptors were involved in the effects of THC and cannabinoid CB1 receptors were involved in the effects of morphine on food reinforced behavior. Thus, activation of both endogenous cannabinoid and opioid systems appears to jointly facilitate motivational effects of food measured under progressive-ratio schedules of reinforcement and this facilitatory modulation appears to critically depend on interactions between these two systems. These findings support the proposed therapeutic utility of cannabinoid agonists and antagonists in eating disorders. PMID- 15812568 TI - Comparison of alterations in c-fos and Egr-1 (zif268) expression throughout the rat brain following acute administration of different classes of antidepressant compounds. AB - The majority of immediate-early gene (IEG) studies focus on a few key brain regions associated with the class of psychoactive compound being studied. Recently, using a meta-analysis of the c-fos literature, we demonstrated the utility of c-fos profiling to classify such compounds. The present study examined acute delivery of a range of antidepressant classes; fluoxetine, imipramine, LiCl, and mirtazapine. The dual aims were to study the IEG profiles of these varying classes of antidepressants throughout the rat brain and to compare the utility of c-fos or Egr-1 as IEGs to classify clinically efficacious antidepressants. All antidepressants increased c-fos mRNA in the central amygdala, as previously shown, while c-fos was also increased in the anterior insular cortex and significantly decreased within the septum. Although acute antidepressant administration altered c-fos expression in a number of brain regions, Egr-1 expression was only significantly altered in the central amygdala, suggesting that Egr-1 may not be as useful a marker to investigate acute antidepressant treatment. The fact that these drugs, including the previously unclassified antidepressant mirtazapine, share a number of common loci of activation, which are implicated by human and animal studies in depression, adds further support to the use of IEG mapping to classify psychoactive compounds. PMID- 15812569 TI - Gestational dexamethasone treatment elicits sex-dependent alterations in locomotor activity, reward-based memory and hippocampal cholinergic function in adolescent and adult rats. AB - Glucocorticoids are the consensus treatment for preventing respiratory distress syndrome in preterm infants but there is emerging evidence of subsequent neurobehavioral abnormalities, independent of somatic growth effects. Pregnant rats were given 0.2 mg/kg of dexamethasone, a dose commensurate with clinical use, on gestational days 17-19 and behavioral evaluations were made on the offspring in adolescence and adulthood. The dexamethasone groups had the same body weights as the controls but nevertheless displayed long-term, sex-selective alterations in locomotor and cognitive behaviors. In the figure-8 activity apparatus, dexamethasone treatment ablated the normal sex differences in locomotor activity by reducing values in females to the lower level typical of males; habituation of activity similarly was impaired in females, reducing the profile to match that of control males, while male rats in the dexamethasone group showed a partially feminized pattern of habituation. In the 8-arm radial maze, control rats displayed typical sex differences, with male rats performing more accurately than females. Dexamethasone treatment eliminated this normal dichotomy, delaying learning in males while improving performance in females to the level normally seen in control males. Finally, we assessed hippocampal [3H]hemicholinium-3 binding as a biomarker for cholinergic synaptic activity, and again found loss of sex differences in the dexamethasone group: values in males were increased to the higher levels typical of females. These results indicate that gestational treatment with dexamethasone obtunds the normal sex differences in neurochemistry and behavior that are typically seen in adolescence in adulthood, thus producing sex-selective alterations in activity, learning, and memory. PMID- 15812570 TI - Acute delta9-tetrahydrocannabinol-induced deficits in reversal learning: neural correlates of affective inflexibility. AB - Despite concerns surrounding the possible adverse effects of marijuana on complex cognitive function, the processes contributing to the observed cognitive deficits are unclear, as are the causal relationships between these impairments and marijuana exposure. In particular, marijuana-related deficits in cognitive flexibility may affect the social functioning of the individual and may contribute to continued marijuana use. We therefore examined the ability of rats to perform affective and attentional shifts following acute administration of Delta(9)-tetrahydrocannabinol (THC), the primary psychoactive marijuana constituent. Administration of 1 mg/kg THC produced marked impairments in the ability to reverse previously relevant associations between stimulus features and reward presentation, while the ability to transfer attentional set between dimensional stimulus properties was unaffected. Concurrent in situ hybridization analysis of regional c-fos and ngfi-b expression highlighted areas of the prefrontal cortex and striatum that were recruited in response to both THC administration and task performance. Furthermore, the alterations in mRNA expression in the orbitofrontal cortex and striatum were associated with the ability to perform the reversal discriminations. These findings suggest that marijuana use may produce inelasticity in updating affective associations between stimuli and reinforcement value, and that this effect may arise through dysregulation of orbitofrontal and striatal circuitry. PMID- 15812571 TI - Group II and III metabotropic glutamate receptors suppress excitatory synaptic transmission in the dorsolateral bed nucleus of the stria terminalis. AB - Conditions such as anxiety, drug abuse, and post-traumatic stress disorder are thought to reflect alterations in central nervous system stress and reward circuitry. Recent evidence suggests a key component of this circuitry is the bed nucleus of the stria terminalis (BNST). In particular, regulation of glutamatergic transmission in the BNST plays a critical role in animal performance on anxiety tasks. Metabotropic glutamate receptors (mGluRs) have been implicated in stress and drug addiction and are known to regulate glutamatergic transmission in many brain regions. We have utilized both extracellular field potential and whole-cell patch-clamp recording in an in vitro slice preparation of mouse dorsal anterolateral BNST to determine whether G(i/o)-linked mGluRs modulate excitatory transmission in this region. We find that activation of group II and group III mGluRs in an in vitro slice preparation of the dBNST causes a depression of excitatory transmission. The depression evoked by group II mGluR activation may represent a form of synaptic plasticity as prolonged activation of the receptor produces a long-term depression of glutamatergic transmission. Based on paired-pulse ratio analysis, initiation of depression by group II and group III mGluR subfamilies appears to, at least in part, involve decreased glutamate release. In total, our data suggest a plausible site of action for some of the anxiolytic effects of group II and group III mGluR agonists. PMID- 15812572 TI - Chronic lithium chloride administration to unanesthetized rats attenuates brain dopamine D2-like receptor-initiated signaling via arachidonic acid. AB - We studied the effect of lithium chloride on dopaminergic neurotransmission via D2-like receptors coupled to phospholipase A2 (PLA2). In unanesthetized rats injected i.v. with radiolabeled arachidonic acid (AA, 20:4 n-6), regional PLA2 activation was imaged by measuring regional incorporation coefficients k* of AA (brain radioactivity divided by integrated plasma radioactivity) using quantitative autoradiography, following administration of the D2-like receptor agonist, quinpirole. In rats fed a control diet, quinpirole at 1 mg/kg i.v. increased k* for AA significantly in 17 regions with high densities of D2-like receptors, of 61 regions examined. Increases in k* were found in the prefrontal cortex, frontal cortex, accumbens nucleus, caudate-putamen, substantia nigra, and ventral tegmental area. Quinpirole, 0.25 mg/kg i.v. enhanced k* significantly only in the caudate-putamen. In rats fed LiCl for 6 weeks to produce a therapeutically relevant brain lithium concentration, neither 0.25 mg/kg nor 1 mg/kg quinpirole increased k* significantly in any region. Orofacial movements following quinpirole were modified but not abolished by LiCl feeding. The results suggest that downregulation by lithium of D2-like receptor signaling involving PLA2 and AA may contribute to lithium's therapeutic efficacy in bipolar disorder. PMID- 15812573 TI - Attraction of two lacewing species to volatiles produced by host plants and aphid prey. AB - It is well documented that host-related odors enable many species of parasitoids and predatory insects to locate their prey and prey habitats. This study reports the first characterization of prey and prey host odor reception in two species of lacewings, Chrysoperla carnea (Say) and Chrysopa oculata L. 2-Phenylethanol, one of the volatiles emitted from their prey's host plants (alfalfa and corn) evoked a significant EAG response from antennae of C. carnea. Traps baited with this compound attracted high numbers of adult C. carnea, which were predominantly females. One of the sex pheromone components (1R,4aS,7S,7aR)-nepetalactol of an aphid species, Acyrthosiphon pisum (Harris) attracted only C. oculata adults. Single sensillum recordings showed that the olfactory neurons of C. carnea responded to both 2-phenylethanol and aphid sex pheromone components, but those of C. oculata only responded to the latter. PMID- 15812574 TI - Novel approaches in the therapy of metastatic renal cell carcinoma. AB - Renal cell carcinoma (RCC) is the most lethal of the common urologic malignancies, with approximately 40% of patients eventually dying of cancer progression. Approximately one third of patients present with metastatic disease, and up to 40% treated for localized disease have a recurrence. Recent advances in the understanding of the pathogenesis, behavior, and molecular biology of RCC have paved the way for developments that may enhance early diagnosis, better predict tumor prognosis, and improve survival for RCC patients. The recent discovery of molecular tumor markers is expected to revolutionize the staging of RCC in the future and lead to the development of new therapies based on molecular targeting. Cytokine-based immunotherapy can be considered standard therapy in the treatment of metastatic RCC today. However, new therapies such as tumor vaccines, anti-angiogenesis agents, and small molecule inhibitors are being developed to improve efficacy and treat those patients who are unable to tolerate or are resistant to systemic immunotherapy. The aim of this review is to provide an update on current therapeutic approaches and targeted molecular therapy for metastatic RCC. PMID- 15812575 TI - CSF-1 induces fos gene transcription and activates the transcription factor Elk-1 in mature osteoclasts. AB - Mice with targeted deletion of the fos gene fail to develop mature osteoclasts, reflecting an absolute requirement for the c-Fos proto-oncogene in osteoclast precursors. C-Fos is also expressed in mature osteoclasts; however, the regulation of fos in these cells has not been studied. By using cultured murine osteoclast-like cells (OCLs) we found that treatment with colony-stimulation factor 1 (CSF-1) induced a 3.9-fold increase in c-Fos rnRNA at 30 minutes and a 2.6-fold increase at 60 minutes. With use of mature osteoclasts isolated from transgenic mice expressing the bacterial Lac-Z gene under the control of the murine fos promoter, we were able to directly demonstrate transcriptional activation of fos by CSF-1 in these cells. Transcriptional activation was 2.6 fold greater at 5 minutes and 2.8-fold greater at 15 minutes in CSF-1-treated cells than in vehicle-treated cells. CSF-1 induced nuclear protein binding to the fos serum response element that was significantly attenuated by antibodies to the transcription factor Elk-1 but not by Sap-1a. Treatment of mature osteoclasts with CSF-1 for 2 hours resulted in a significant increase in the levels of nuclear c-Fos protein. These data demonstrate that CSF-1 upregulates c-fos expression in mature osteoclasts at least in part via transcriptional activation of fos. CSF-1 induced binding of Elk-1 to the fos gene serum response element appears to be part of the molecular mechanism by which this occurs. PMID- 15812576 TI - Mitigation of ectopic calcification in osteopontin-deficient mice by exogenous osteopontin. AB - Ectopic calcification is a major cause of bioprosthetic heart valve failure. New therapeutic opportunities are offered by the growing understanding that ectopic calcification is an actively regulated process involving several key gene products. One of these products, osteopontin (OPN), is a glycosylated phosphoprotein previously shown to inhibit apatite crystal formation, induce carbonic anhydrase II, and promote mineral resorption. In this study, OPN deficient mice (OPN-/-) were utilized as an in vivo model to stimulate the ectopic calcification of glutaraldehyde-fixed bovine pericardium (GFBP) tissue and to examine OPN delivery and structure-function relationships with respect to its anti-calcific activity. Significant calcification of GFBP tissue was obtained within 7 days of subcutaneous implantation in OPN-/- mice. Direct rescue of the calcification phenotype was achieved by the administration of exogenous recombinant rat, histidine-fused OPN (rat His-OPN) to the implant site via soluble injection (up to 72% mitigation achieved) or adsorption onto the implant materials (up to 91% mitigation achieved). Effects were specific, since neither fibronectin nor polyhistidine alone could mitigate calcification of GFBP. The maximum anti-calcific effect was achieved only when rat His-OPN was adequately phosphorylated and contained a functional arginine-glycine-aspartate (RGD) cell adhesive domain. Furthermore, CAII levels in host cells surrounding GFBP were greatest when phosphorylated, RGD-containing rat His-OPN was adsorbed. These data suggest that both physical inhibition, mediated by phosphorylation sites in OPN, as well as the induction of CAII and mineral regression, mediated by the RGD domain, contribute to the unique ability of OPN to mitigate ectopic calcification of bioprosthetic valve tissue. PMID- 15812577 TI - Changes in osteoprotegerin/RANKL system, bone mineral density, and bone biochemicals markers in patients with recent spinal cord injury. AB - This study analyzed the temporal and regional variations in bone loss and explored bone cell activities via biochemical markers during an extended follow up in patients with spinal cord injury (SCI). In parallel, the possible role of the osteoprotegerin (OPG)/RANKL system in disuse osteoporosis was investigated. Seven male patients with acute and complete SCI (31.3 +/- 9.5 years) and 12 able bodied (AB) men (26.9 +/- 4.2 years) participated in the study. Measurements were performed 16, 24, 36, 48, and 71 weeks after injury. At week 16, marked calcium homeostasis disturbance and a concomitant increase in bone resorption markers were observed, reflecting an intense bone degradation process. Resorption activity decreased continuously with time. Contrasting with the great rise in the resorption markers, the bone formation markers showed little variation. During the period of investigation, a loss in bone mineral density (BMD) was demonstrated for the total body (-4.3%), pelvis (-15.7%) and lower limbs ( 15.2%), whereas BMD did not change at the lumbar spine, upper limbs, or skull. At all stages, SCI patients had lower serum RANKL levels and higher serum OPG levels than did AB controls, but no significant variation with time was observed for either cytokine. These findings suggest that bone resorption persisted long after SCI and specifically affected BMD at sublesional sites. The marked modification of serum OPG/RANKL levels in SCI patients suggests that this system is affected, in disuse osteoporosis. However, the precise biologic role of the OPG/RANKL system in the bone tissue of SCI patients has yet to be determined. PMID- 15812578 TI - Osteoblast responses one hour after load-induced fluid flow in a three dimensional porous matrix. AB - When bone is loaded, substrate strain is generated by the external force and this strain induces fluid flow that creates fluid shear stress on bone cells. Our current understanding of load-driven gene regulation of osteoblasts is based primarily on in vitro studies on planer two-dimensional tissue culture substrates. However, differences between a flat layer of cells and cells in 3 dimensional (3D) ECM are being recognized for signal transduction. Proliferation and differentiation of osteoblasts are affected by substrate geometry. Here we developed a novel 3D culture system that would mimic physiologically relevant substrate strain as well as strain-induced fluid flow in a 3D porous collagen matrix. The system allowed us to evaluate the responses of osteoblasts in a 3D stress-strain environment similar to a mechanical field to which bone is exposed. Using MC3T3-E1 osteoblasts grown in the 3D collagen matrix with and without hydroxyapatite deposition, we tested the role of strain and the strain-induced fluid flow in the expression of the load-responsive genes such as c-fos, egr1, cox2, osteopontin, and mmp1B involved in transcriptional regulation, osteogenesis, and rearrangement of ECM. Strain-induced fluid flow was visualized with a microspheres approximately 3 microm in diameter in real time, and three viscoelastic parameters were determined. The results obtained by semi quantitative PCR, immunoblot assay, enzymatic activity assays for collagenase and gelatinase, and mechanical characterization of collagen matrices supported the dominant role of strain-induced fluid flow in expression of the selected genes one hour after the mechanical treatment. PMID- 15812579 TI - Long-term morbidity and mortality after a clinically diagnosed vertebral fracture in the elderly--a 12- and 22-year follow-up of 257 patients. AB - The objective of this study was to analyze the long-term morbidity and mortality in patients with a clinically diagnosed vertebral fracture. Seventy men with a mean age of 70 years (range 50-91 years) and 187 women with a mean age of 72 years (range 50-96 years) were radiographically diagnosed as having a vertebral fracture in the thoracic or lumbar spine at the Malmo University Hospital (Sweden) during 1979. At the time of a follow-up examination 12 years later, 56 of the 76 patients who were still alive participated in an investigation that evaluated back pain and subjective health status by a questionnaire. Forty-four of these subjects also participated in a further radiologic examination of the spine. Serving as controls were age- and gender-matched subjects from the Malmo cohort of the European Vertebral Osteoporosis Study (EVOS). A mortality analysis was also conducted, covering 22 years following the baseline fracture. There were more female patients, who, in comparison with the controls, 12 years after the diagnosis, had had back pain during the year preceding the follow-up (72% vs 33%, P < 0.001), had current back pain (42% vs. 19%, P = 0.006), and had a subjectively impaired health status (44% vs. 17%, P < 0.001). The corresponding differences in men reached only a borderline significance, for both back pain during the year preceding the follow-up (60% vs. 28%, P = 0.07) and current back pain (40% vs. 15%, P = 0.09), whereas there was no difference in subjective health status. The incidence of new vertebral fractures in individuals with a clinically diagnosed vertebral fracture during the following 12 years was in men 25 per 1,000 person-years and in women 49 per 1000 person-years. There were more women with a new vertebral fracture at the 12-year follow-up examination who, in comparison with women without a new fracture, had had back pain during the year preceding the follow-up examination (90% vs. 50%, age-adjusted P = 0.02) and had current back pain (65% vs. 21%, age-adjusted P = 0.03). Women with a new vertebral fracture at the 12-year follow-up examination had a higher subsequent mortality rate in the next 10 years [age-adjusted hazard ratio 2.8 (95% CI 1.0 7.9)] as compared with women without. The mortality rate during the 22 years following the diagnosis among the male patients was 111.7 per 1,000 person-years as compared with 73.4 per 1,000 person-years among the male population at risk. The mortality rate among the female patients was 95.1 per 1,000 person-years as compared with 62.0 per 1,000 person-years among the female population at risk. We conclude that a clinically diagnosed thoracic or lumbar vertebral fracture in the elderly can be regarded as a risk factor for subsequent, long-term morbidity, especially in women, and for mortality in both genders. PMID- 15812580 TI - Effects of pleiotrophin (PTN) over-expression on mouse long bone development, fracture healing and bone repair. AB - Pleiotrophin (PTN) was found to have potent effects on regulation of osteoblast recruitment, proliferation and differentiation. The present study examined the long-term effects of targeted PTN over-expression on bone development and repair in a transgenic mouse model. Femurs and tibiae from the PTN transgenic mice and the wild type mice at age 1, 2, 4, 6, 12 and 24 months were collected, and examined by radiography, peripheral quantitative computed tomography (pQCT), histology and mechanical testing. Age-matched PTN and the control mice received a standardized femoral fracture, followed by regular x-rays and sacrificed at day 16 post-fracture for histology examination. A cortical hole was drilled on the tibiae of age-matched PTN and wild type mice, collagen sponge with either saline, 100 ng of rhBMP-2 or rhPTN was implanted in the holes, and animals were sacrificed 10 days later, subject to pQCT and histology examinations. During early stages of bone development, the PTN mice had advanced bone growth in length and maturation, but the difference diminished in later life. The fracture healing was impaired in the PTN mice, and there was delayed callus formation and remodelling. The cortical holes treated with BMP-2 in the PTN mice had significantly less trabecular bone formation. The current study confirmed that the targeted PTN over-expression in mouse bone has moderate enhancing effects on early bone development; but the bones become brittle in later life. Fracture healing was impaired in the adult PTN mice and this may be due to inhibitory effects of PTN over-expression on BMP-2 mediated bone induction. PMID- 15812581 TI - Osteoprotegerin treatment impairs remodeling and apparent material properties of callus tissue without influencing structural fracture strength. AB - The influence of osteoprotegerin (OPG) treatment on callus formation, callus tissue structural strength, apparent material properties, and histology of tibia fractures in rats was investigated after 3 weeks and 8 weeks of healing. OPG was given intravenously (10 mg/kg twice weekly) during the entire observation period, and control animals with fractures received vehicle only. When compared with control fractures after 3 weeks of healing, OPG treatment reduced the number of osteoclasts in the callus tissue (93%, P < 0.001) and hampered resorption of genuine cortical bone in the fracture line; OPG treatment did not influence callus dimensions, callus bone mineral content (BMC), fracture structural strength, or callus tissue apparent material properties. When compared with control fractures after 8 weeks of healing; OPG treatment reduced the number of osteoclasts in callus tissue (92%, P < 0.001), augmented callus dimensions (anteriorposterior diameter: 12%, P = 0.034, mediolateral diameter: 13%, P = 0.013), and increased callus BMC (50%, P = 0.007); OPG treatment hampered deposition of new woven bone at the fracture line of the genuine cortical bone (new woven bone present in all vehicle animals, but only in 13% of the OPG treated animals (P < 0.001)); OPG treatment did not influence structural strength of the fractures, but decreased apparent material properties of the callus tissue (ultimate stress: 51%, P < 0.001; elastic modulus: 42%, P = 0.033). The experiment demonstrates that OPG treatment does not influence the early callus expansion and fracture strength. However, during the subsequent period of remodelling, OPG treatment impairs the normal remodeling and consolidation processes. PMID- 15812582 TI - Screening elderly women for risk of future fractures--participation rates and impact on incidence of falls and fractures. AB - We have assessed the acceptability of a method for screening for risk of future hip fracture in elderly women. After receipt of an initial response to a mailed risk-factor questionnaire sent out to 5,306 women, women were randomly assigned to active or control groups. The active group was invited to participate in a screening visit that comprised a life-style questionnaire and a quantitative ultrasound heel scan. General practitioners (GPs) of women who were found to be in the lowest quartile of broadband ultrasound attenuation and/or who had two or more risk factors for hip fracture were advised to prescribe a calcium and vitamin D supplement. A second mailed questionnaire was sent to both groups 1 to 3 years later. Compared with the control group, the active group had a 56% lower risk of fracture (odds ratio [OR], 0.44; 95% confidence interval [CI], 0.24-0.81 adjusted age, weight, and treatment status). At follow-up, the proportion of fallers in the active group (25.3%) was lower than that in the control group (29.6%) (P = 0.064). The control group was found to have a higher rate of falls at follow-up than the active group (95% CI, 0.02-0.22); no difference was found at baseline (95% CI, -0.08 to +0.14). The screening method used was found to be acceptable to the majority of elderly women in this study. Screening the elderly in this way together with simple advice on treatment appears to reduce the age associated increase in fall rates and the number of subsequent fractures. This form of screening may provide a cost-effective method to reduce falls and fractures in free-living elderly women. However, no such cost-effectiveness analysis has been performed to date. PMID- 15812583 TI - Relationship between surgeon caseload and sphincter preservation in patients with rectal cancer. AB - PURPOSE: The aim of this study was to determine by means of a national database whether higher surgeon caseload correlates with greater utilization of sphincter sparing procedures than of abdominoperineal resections in treatment of patients with rectal cancer. METHODS: Patients with a primary International Classification of Diseases-9 diagnosis code of rectal cancer who underwent a sphincter-sparing procedure or abdominoperineal resection were selected from the 1997 Nationwide Inpatient Sample, a database that represents 20 percent of all U.S. community hospital discharges. Multivariable logistic regression models were used on a 20 percent sample of this database to estimate the risk-adjusted relationship between surgeon caseload volume and the odds of receiving a sphincter-sparing procedure. All models were adjusted for age, gender, race, hospital region, and patient comorbidity. RESULTS: The study population (n = 477) was 70.4 percent white and 57.9 percent male with an average age of 67.6 years. The mean Deyo comorbidity score was 7.0. Patients treated by surgeons in the highest-volume category (> or =10 rectal cancer surgeries per year) compared with those treated by surgeons in the lowest-volume category (1-3 rectal cancer surgeries per year) were significantly more likely to undergo a sphincter-sparing procedure, after adjustment for other covariates (odds ratio = 5.05; 95 percent confidence interval, 2.5-10.22). CONCLUSION: This analysis suggests that rectal cancer patients treated by high-volume surgeons are five times more likely to undergo sphincter-sparing procedures than those treated by low-volume surgeon. This has significant implications for those seeking a sphincter-preserving option for the treatment of their rectal cancer. PMID- 15812584 TI - Long-term results of the anterior Delorme's operation in the management of symptomatic rectocele. AB - PURPOSE: Although the results of surgery for symptomatic rectocele seem satisfactory initially, there is a trend toward deterioration with time. This study was designed to assess the long-term outcome of Anterior Delorme's operation for rectocele. METHODS: Questionnaires were sent to all females who had Anterior Delorme's operation performed in Auckland between 1990 and 2000. The questionnaires included obstructed defecation symptoms and a validated fecal incontinence severity index questionnaire and fecal incontinence quality of life questionnaire. Preoperative and postoperative obstructed defecation symptoms and incontinence score were compared. RESULTS: A total of 150 females (mean age, 56 (range, 30-83) years) who had an Anterior Delorme's operation for a rectocele were identified. One hundred seven patients (71.5 percent; mean age, 56 years) completed the questionnaire. Median follow-up was four (range, 2-11) years. The number of patients with obstructed defecation reduced from 87 preoperatively to 23 postoperatively using Rome II criteria (P < 0.0001). Postoperatively there was a reduction in the number of patients with each of the symptoms of obstructed defecation from 83 to 27 for straining, 87 to 33 for incomplete emptying, 64 to 14 for feeling of blockage, 41 to 10 for digitation (P < 0.0001 for all). The median incontinence score reduced from 20 of 61 preoperatively to 12 of 61 postoperatively (P = 0.0001). CONCLUSIONS: In patients with symptomatic rectocele, Anterior Delorme's operation provides long-term benefit for patients with obstructed defecation and leads to a significant improvement of incontinence scores. PMID- 15812585 TI - Gonyautoxin: new treatment for healing acute and chronic anal fissures. AB - PURPOSE: The mayor symptoms of chronic anal fissure are permanent pain, intense pain during defecation that lasts for hours, blood in the stools, and sphincter cramps. It is subsequent to formation of fibrosis infiltrate that leads to an increased anal tone with poor healing tendency. This vicious circle leads to fissure recurrence and chronicity. This study was designed to show the efficacy of gonyautoxin infiltration in healing patients with anal fissures. METHODS: Gonyautoxin is a paralyzing phytotoxin produced by dinoflagellates. Fifty recruited patients received clinical examination, including proctoscopy and questionnaire to evaluate the symptoms. Anorectal manometries were performed before and after toxin injection. Doses of 100 units of gonyautoxin in a volume of 1 ml were infiltrated into both sides of the anal fissure in the internal anal sphincter. RESULTS: Total remission of acute and chronic anal fissures were achieved within 15 and 28 days respectively. Ninety-eight percent of the patients healed before 28 days with a mean time healing of 17.6 +/- 9 days. Only one relapsed during 14 months of follow-up. Neither fecal incontinence nor other side effects were observed. All patients showed immediate sphincter relaxation. The maximum anal resting pressures recorded after two minutes decreased to 56.2 +/- 12.5 percent of baseline. CONCLUSIONS: Gonyautoxin breaks the vicious circle of pain and spasm that leads to anal fissure. This study proposes gonyautoxin anal sphincter infiltration as safe and effective alternative therapeutic approach to conservative, surgical, and botulinum toxin therapies for anal fissures. PMID- 15812586 TI - Digital rectal examination of sphincter pressures in chronic anal fissure is unreliable. AB - PURPOSE: Chronic anal fissure is said to be associated with internal sphincter hypertonia. However, an unknown proportion of fissures may be associated with normal or even low resting pressures and may subsequently be resistant to pharmacological treatments or at risk from surgical treatments, both of which aim to reduce sphincter hypertonia. This study investigated the ability of surgeons to detect low or normal pressure fissures by digital rectal examination. METHODS: Patients with chronic anal fissure were assessed prospectively. The results of anal manometry performed on these patients were compared with digital rectal assessment of sphincter tone undertaken by a surgeon blinded to the manometry results. RESULTS: Forty consecutive patients (21 male) with chronic anal fissure were studied. Twenty-two (55 percent) had normal maximum resting pressure and a further 3 (8 percent) had low pressures on anal manometry. On clinical assessment, only five (13 percent) patients were evaluated as having no anal hypertonia. Clinical assessment of anal tone correctly identified 14 of 15 patients with high manometric maximum resting pressure (sensitivity, 93 percent), yet detected only 4 of 25 patients with normal or low pressures (specificity, 16 percent). The positive predictive value of clinical assessment of anal tone was 40 percent and the negative predictive value, 80 percent. CONCLUSIONS: The incidence of patients with chronic anal fissure without high manometric maximum resting pressure is higher than previously reported. The ability of surgeons to identify this group clinically was poor. It is reasonable to treat all patients primarily medically, and then selectively investigate by manometry those patients who fail medical therapy before considering lateral sphincterotomy. PMID- 15812587 TI - Lymphatic staging in colorectal cancer: pathologic, molecular, and sentinel node techniques. AB - PURPOSE: Accurate staging in colorectal cancer is important to predict prognosis and identify patients who could benefit from adjuvant therapy. Patients with lymphatic metastasis, Stage III/Dukes C, are generally treated with adjuvant chemotherapy. Still, patients without lymphatic metastasis do have relapse as high as 27 percent in five years in Dukes B2. It is hypothesized that these patients have occult (micro)metastasis in their lymph nodes. If these (micro)metastasis can be identified, these patients might benefit from adjuvant therapy. We reviewed the literature on procedures to improve lymph node staging. METHODS: An extensive literature search was performed in PubMed (www.pubmed.com). Using the reference lists, more articles were found. RESULTS: We found 30 articles about sentinel node in colorectal cancer describing original series. Some groups reported several studies including the same patients. We reported their largest studies. For all other techniques, we only included key articles. CONCLUSIONS: Many techniques to improve staging have been described. The finding of occult (micro)metastasis is of prognostic significance in most studies. The sentinel node technique has been recently described for use in colorectal cancer. Although it seems clear that this technique has prognostic potential, it is not yet been shown in a follow-up study. Furthermore, the finding of occult (micro)metastasis in any technique used has not been shown to be clinically significant. Whether to treat patients with adjuvant therapy if occult (micro)metastasis are found needs to be proven in future studies. PMID- 15812588 TI - Lymph node metastases of prostatic adenocarcinoma in the mesorectum in patients with adenocarcinoma or villous tumor of the rectum with collision phenomenon in a single lymph node: report of five cases. AB - PURPOSE: Lymph node involvement is the most important prognostic factor when staging patients with rectal cancer. Cancer originating from sites other than rectum rarely may metastasize to the mesorectum. We report five patients with metastatic prostatic carcinoma to mesorectal lymph nodes, with the "collision phenomenon" in one lymph node. The diagnosis of prostate cancer was clinically unsuspected in two cases. METHODS: We examined three cases of primary adenocarcinoma and two villous tumors with high-grade dysplasia (patient age range, 52-74 (mean, 63) years) of the middle or lower third of the rectum. All patients underwent low anterior rectal resection with total mesorectal excision and colorectal or coloanal anastomosis. We used a manual technique for lymph node detection after overnight fixation in 10 percent formalin. All lymph nodes identified (range, 15-32; mean, 21 nodes per patient) were examined histologically. RESULTS: Of 106 lymph nodes examined, 20 contained metastases: 9 from rectal adenocarcinoma, 10 from prostatic adenocarcinoma, and 1 with metastatic foci from both tumors. The diagnosis of prostatic carcinoma was readily confirmed by immunostaining for prostatic-specific antigen, and prostatic acid phosphatase. CONCLUSIONS: Mesorectal lymph node dissection provides prognostic information in rectal cancer, but careful examination may reveal other unsuspected pathology. Immunohistochemical staining is an essential tool in distinguishing the origin of a lymph node metastasis, especially when the histology does not look typical for rectal carcinoma. Moreover, these observations highlight the connection that can exist between mesorectal lymph node drainage, and extra mesorectal lymph nodes drainage. PMID- 15812589 TI - Yersinia enterocolitica infection in a patient with hemachromatosis masquerading as proximal colon cancer with liver metastases: report of a case. AB - A patient with genetic hemachromatosis presented with a clinical picture suggesting malignancy and CT evidence of lesions in the right colon and liver. Colonoscopy failed to confirm the suspected diagnosis. Blood and stool cultures were positive for yersinia enterocolittica infection. This case illustrates the need to confirm clinically and radiologically suspected malignancy. It also serves as a model of how localized gastrointestinal pathology can result from the interaction of host genetic factors and specific microbial species. PMID- 15812590 TI - Hyponatremia and seizures after bowel preparation: report of three cases. AB - Oral sodium phosphate and sodium picosulfates/magnesium citrate are commonly used to evacuate the colon and rectum before colonoscopy or colorectal surgery. These substances, however, are known to cause electrolyte abnormalities. Seizures caused by electrolyte abnormalities associated with bowel preparation have only rarely been reported. We report the cases of three patients with no prior history of seizures, who had their first seizure associated with hyponatremia following ingestion of sodium phosphate or sodium picosulfates/magnesium citrate combination. Care must be taken with patients with a low seizure threshold and those with possible chronic sodium depletion, such as patients on thiazide diuretics, who are undertaking bowel preparation with oral sodium phosphate or sodium picosulfates/magnesium citrate combination. PMID- 15812593 TI - Surgical treatment of cerebellopontine angle meningiomas in elderly patients. AB - BACKGROUND: The aim of this study is to assess the morbidity and mortality of meningioma surgery in patients over 70 years of age harbouring a tumour at the cerebellopontine angle as one representative location of the posterior fossa in comparison with a matched group of young patients. METHOD: A retrospective analysis based on clinical charts, surgical records, histological records, imaging studies and follow up records was conducted to select patients over 70 years who underwent surgery for cerebellopontine angle meningiomas. Tumours with comparable size and location were matched with the younger group. FINDINGS: There were 421 meningiomas located in the cerebellopontine angle, 21 patients were older than 70 years (range 70-84). Median Karnofsky-Index at presentation was 80 (50-90), 16 patients had a physical status grading ASA 2 and 5 patients ASA 3. The average length of hospital stay was 22 days (7-99 days). The postoperative median Karnofsky score at time of discharge was 80 (50-90). The most common medical complication was postoperative pneumonia in 4 patients, among them 3 patients had lower cranial nerve disturbances postoperatively. There were 56 younger patients (mean age 52.4 years; range 24.5-69.75 years) with corresponding tumour size and location. Pre-op Karnofsky score was 80 (70-90), 53 patients were graded as ASA 2 and 3 patients as ASA 3. Length of hospital stay was 13.6 days (8 32 days). Post-op Karnofsky score was 80 (50-90). Among 5 patients with postoperative lower cranial nerve disturbances no patient had pneumonia postoperatively. There was no peri-operative mortality in either group. CONCLUSIONS: With modern neurosurgical techniques and neuro-anesthesia elderly patients with CPA meningiomas can be operated on with acceptable low morbidity and good neurological outcome but recovery from surgery lasts longer compared to younger patients. However, postoperative lower cranial nerve deficits in elderly patients may not be well tolerated compared with younger patients. PMID- 15812594 TI - Rac2 expression and mutation in human brain tumors. AB - BACKGROUND: Rac1 and Rac2 are interchangeable in NADPH oxidase activation. Rac1 plays an important role in regulating nuclear signalling and in the activation of transcriptional factors that regulate gene expression and cell growth. Our previous study observed mutation in effector region of Rac1 gene in brain tumors. Little is known about the expression and mutation of Rac2 in human brain tumors. METHOD: We examined the expression of Rac2 by using reverse transcriptase polymerase chain reaction (RT-PCR) and northern blot analysis and the mutation of Rac2 gene by using DNA sequence analysis. FINDINGS: The decreased expression of Rac2 was found in 15 cases (57.7%) including 8 of 10 astrocytomas, 2 of 8 meningiomas, and 5 of 8 pituitary adenomas. Two of 13 cases with decreased expression of Rac2 had gene mutation. Only two of 26 cases had Rac2 overexpression in which no Rac2 gene mutation was found. Four of 8 cases with normal Rac2 expression had Rac2 gene mutation. The site of Rac2 gene mutation had no hot spots and was not concentrated in the effector region. CONCLUSIONS: Our results showed a low frequency of mutation and no hot spots of mutation in Rac2 gene in brain tumors, suggesting a decreased possibility of Rac2 in the brain tumorigenesis. The role of high frequency of decreased Rac2 expression in brain tumors, particularly in malignant astrocytomas, needs further investigations to be elucidated. PMID- 15812595 TI - Microsurgical anatomy of the lateral walls of the pituitary fossa. AB - BACKGROUND: The aim was to evaluate the microanatomy of the lateral wall of the pituitary fossa in cadavers. METHODS: Histological sections of sellar-parasellar specimens from 13 cadaver heads were examined. The thickness of the pituitary capsule and inferior and lateral walls of the pituitary fossa were measured, and the collagenous structure of these layers was evaluated. FINDINGS: The pituitary gland is enveloped by a tough, thin, fibrous capsule. The inferior wall of the pituitary fossa is composed of relatively thick dura (mean thickness in the 13 specimens, 171 microm). Each lateral wall of the fossa has a thin layer of dura (mean thickness in the specimens, 85 microm). The pituitary capsule and the dural layers in the lateral and inferior walls of the fossa were immunopositive for collagen types I and II. Collagen types III, IV and V were detected only in the pituitary capsule. CONCLUSIONS: Weakness of the lateral walls of the pituitary fossa and the degree to which collagen fibres in the pituitary capsule have been biochemically damaged are important factors in infiltration of the cavernous sinus by a pituitary adenoma. PMID- 15812596 TI - Radiofrequency thermal ablation for recurrent meningioma extending extracranially. AB - BACKGROUND: Image-guided and temperature-controlled radiofrequency thermal ablation techniques were applied to reduce tumor volume and relieve the symptoms caused by extracranial extension of recurrent meningioma. METHOD: We treated two patients with recurrent meningioma, an 81-year-old woman presenting with bulging of the temple and a 68-year-old woman presenting with visual disturbance, facial disfigurement, and sensory disturbance. Neuroimaging in both patients, revealed a large tumor extending extracranially and involving the infratemporal fossa. To avoid injury to important anatomical structures either compressed or entrapped by the tumor, the spatial relation between the planned ablation volume and these structures was confirmed by 3-D reconstruction of the ablation target. During the ablation procedure, local temperatures over the tissue being cauterized were continuously monitored to limit the ablation area to that within the planned volume adjusting RF power. FINDING: Radiofrequency ablation produced tumor necrosis as planned without adverse effects and resulted in swift relief of symptoms and signs with shrinkage of the tumor. CONCLUSION: This technique may be an effective alternative for recurrent meningiomas extending extracranially and for which radical surgical procedures are not indicated. PMID- 15812597 TI - Intracranial aneurysms in patients 18 years of age or under, are they different from aneurysms in adult population? AB - BACKGROUND: Intracranial aneurysms are extremely uncommon in the first two decades of life. This study was undertaken to assess the clinicoradiological features and surgical outcome of intracranial aneurysms in patients less than or equal to 18 years of age; and, to highlight the differences between these and intracranial aneurysms seen in adult patients. METHODS AND MATERIAL: Twenty-two patients, 18 years of age or under (male:female ratio=1.75:1; mean age 14.18+/ 3.8 years, age range 5 to 18 years) and 451 adult patients aged older than 18 years (male:female ratio=1:1.05; mean age 48.21+/-12.71 years, age range, 19 to 81 years) were treated for intracranial aneurysms at our center between January 1991 and July 2003. The univariate statistical analysis was used to compare differences between the two groups. FINDINGS: The patients under 18 years constituted 4.6% of the total patient population having intracranial aneurysms. The incidence of associated medical diseases was greater in patients under 18 years than in the adults (9% versus 0.26%, p<0.05). The incidence of seizures was more than double in patients under 18 years (36% versus 17%, p<0.05). The incidence of intracerebral haematoma (ICH; 41% versus 22.5%, p>0.05), intraventricular haemorrhage (IVH; 45% versus 34%, p>0.05), and hydrocephalus (36% versus 25%, p>0.05) were higher in patients under 18 years. In adult patients, anterior communicating artery (AcoA) and in children, ICA bifurcation were the most frequent sites of aneurysm formation respectively (p<0.05). The incidence of giant aneurysms was nearly double in children (13.6% versus 6.5%, p>0.05). The incidence of clinical vasospasm was almost the same in both groups. The overall outcome was favourable in 82% of patients under 18 years and 58.8% in adults. The management mortality in patients under 18 years was 9.1%, while in the adult patients, it was 19%. CONCLUSION: In patients under 18 years of age, there was a definite male predominance; a higher incidence of seizures; and, the ICA bifurcation formed the most frequent site of intracranial aneurysms. In adults, AcoA a was the commonest site. Rebleeding and delayed ischaemic deficits were the major causes of morbidity. Favorable outcome after surgery in young patients was better in comparison to their adult counterparts. PMID- 15812598 TI - Intermittently administered parathyroid hormone 1-34 reverses bone loss and structural impairment in orchiectomized adult rats. AB - Male osteoporosis is emerging as a central theme in bone research. As in females, hypogonadism appears as a principal risk factor in men that leads to bone loss and increased fracture incidence. Intermittently administered parathyroid hormone (PTH) reverses bone loss in sex hormone-deprived women and female animals and increases bone mass in elderly men and normal male animals. This study was carried out to assess whether the PTH anabolic activity is also effective in adult castrated males and to gain insight into the underlying tissue processes. Bilateral orchiectomy (ORX) or sham-ORX was performed in 13-week old rats. Five weeks later, the ORX rats were treated intermittently with human PTH(1-34), 80 microg/kg/day or vehicle for 6 weeks. Femora were evaluated by quantitative micro computed tomography followed by dynamic histomorphometry. The trabecular bone volume density showed 40% and 56% ORX-induced loss in the distal metaphysis at 6 weeks and 12 weeks post-ORX, respectively. PTH(1-34) induced supraphysiologic recovery of this bone loss (155% recovery) consequent to a vast increase in trabecular thickness (174% over sham-ORX controls) and a partial reversal (62%) of the decrease in trabecular number. As compared with the results in 12-week, orchiectomized vehicle-administered rats, the PTH(1-34) treatment induced a significant decrease in osteoclast number (20%) and twofold increase in bone formation rate. While ORX did not affect the femoral diaphysis, PTH(1-34) induced marked cortical thickening via the stimulation of endosteal mineral appositional rate (154% over ORX rats). These data portray PTH(1-34) as a highly potent bone anabolic agent in adult ORX rats, mainly by increasing both the trabecular and cortical thicknesses through its effect on osteoblasts and osteoclasts. The adult ORX rat is useful for investigating the processes involved in bone anabolic activity in castrated osteoporotic males and for the development of bone anabolic agents for treating this condition. PMID- 15812599 TI - Osteoporosis assessment by whole body region vs. site-specific DXA. AB - The ability of regional data from whole body scans to provide an accurate assessment of site-specific BMD, osteoporosis prevalence and fracture risk has not been fully explored. To address these issues, we measured total body (TBBD) and site-specific BMD in an age-stratified population sample of 351 women (21-93 years) and 348 men (22-90 years). We found an excellent correlation between AP lumbar spine and total body lumbar spine subregion BMD (r2=0.92), but weaker ones for total hip compared to pelvis region (r2=0.72) or between total wrist and left arm subregion from the whole body scan (r2=0.83). The error in estimating site specific BMD from total body regions ranged from 4.3% (lumbar spine) to 11.2% (femoral neck) in women and from 4.9 to 11.1%, respectively, in men. Site specific versus regional measurements at the lumbar spine and total hip/pelvis provided comparable overall estimates of osteoporosis prevalence, but disagreed on the status of individuals; measurements at whole body regions underestimated osteoporosis as assessed at the femoral neck or total wrist. All measurements were associated with a history of various fractures [age adjusted odds ratios (OR), 1.3 to 2.1 in women and 1.2 to 1.5 in men] and were generally interchangeable, but femoral neck BMD provided the best estimate of osteoporotic fracture risk in women (OR, 2.9; 95% CI, 1.7-5.0). Although there are strong correlations between BMD from dedicated scans of the hip, spine and distal forearm and corresponding regions on the whole body scan, the measurements provide somewhat different estimates of osteoporosis prevalence and fracture risk. PMID- 15812600 TI - A systematic review of research findings on the efficacy of interpersonal therapy for depressive disorders. AB - OBJECTIVE: Interpersonal psychotherapy (IPT) is a time-limited psychotherapy for major depression. The aim of this study is to summarize findings from controlled trials of the efficacy of IPT in the treatment of depressive spectrum disorders (DSD) using a meta-analytic approach. METHODS: Studies of randomized clinical trials of IPT efficacy were located by searching all available data bases from 1974 to 2002. The searches employed the following MeSH categories: Depression/ Depressive Disorder; Interpersonal therapy; Outcome/Adverse Effects/Efficacy; in the identified studies. The efficacy outcomes were: remission; clinical improvement; the difference in depressive symptoms between the two arms of the trial at endpoint, and no recurrence. Drop out rates were used as an index of treatment acceptability. RESULTS: Thirteen studies fulfilled inclusion criteria and four meta-analyses were performed. IPT was superior in efficacy to placebo in nine studies (Weight Mean Difference (WMD) - 3.57 [-5.9, -1.16]). The combination of IPT and medication did not show an adjunctive effect compared to medication alone for acute treatment (RR 0.78 [0.30, 2.04]), for maintenance treatment (RR 1.01 [0.81, 1.25]), or for prophylactic treatment (RR 0.70 [0.30, 1.65]). IPT was significantly better than CBT (WMD -2.16 [-4.16,-0.15]). CONCLUSION: The efficacy of IPT proved to be superior to placebo, similar to medication and did not increase when combined with medication. Overall, IPT was more efficacious than CBT. Current evidence indicates that IPT is an efficacious psychotherapy for DSD and may be superior to some other manualized psychotherapies. PMID- 15812602 TI - Olfactory impairment in monozygotic twins discordant for schizophrenia. AB - OBJECTIVE: Several studies demonstrated olfactory dysfunction in patients with schizophrenia, some reported deficient olfaction in unaffected relatives of schizophrenics as well. This study differentially assessed olfactory acuity as well as smell identification and smell discrimination in monozygotic twins discordant for schizophrenia and healthy, monozygotic control twins, to determine the genetic basis of different olfactory modalities and their association to schizophrenia. METHOD: The Sniffin'Sticks test,a comprehensive and standardized olfactory test, was employed to assess the olfactory function of 10 monozygotic twin pairs discordant for schizophrenia versus 10 age- and sexmatched healthy,monozygotic twin pairs. RESULTS: Olfaction of affected monozygotic twins was globally impaired. Partial olfactory impairment of their unaffected co-twins may point to a genetic cause of olfactory impairment in schizophrenia. The influence of genetic factors was most evident for olfactory acuity and least evident for smell identification. All olfactory functions declined with duration of illness. Side of stimulus presentation did not influence olfactory performance. CONCLUSIONS: Genetic factors associated with olfactory dysfunction may contribute to schizophrenia. The degree of the genetic influence on olfaction depends on the olfactory domain under examination. PMID- 15812601 TI - Antidepressive effects of traditional and second generation antipsychotics: a review of the clinical data. AB - For a long time,in the context of depressive symptoms in schizophrenia traditional neuroleptics were mostly discussed with respect to possible depressiogenic side effects, although some studies argued that they may also have certain antidepressive effects. However, this was not proven at that time in placebo-controlled studies. Placebo-controlled studies performed in recent years have shown that second generation antipsychotics have antidepressive effects which are significantly stronger than those of the traditional neuroleptics. In addition, it was demonstrated that this antidepressive effect can only partially be explained as being secondary to the improvement of positive and negative symptoms, and is apparently predominantly due to a direct (primary) effect on depressive symptoms. It is of special relevance in this context that the antidepressive effect of second generation antipsychotics was recently demonstrated in depression. The positive results from some studies in bipolar depression are especially impressive and underline the antidepressive potencies of novel antipsychotics beyond the spectrum of schizophrenia. PMID- 15812603 TI - Visual emotional stimuli modulation of auditory sensory gating studied by magnetic P50 suppression. AB - The auditory sensory gating system modulates its sensitivity to incoming stimuli and prevents higher brain functions from sensory overload in the primary auditory cortex. We investigated whether visually evoked emotional stimuli affect auditory sensory gating. Magnetic P50 (P50m) suppression was evaluated by magnetoencephalography in fifteen healthy subjects while they viewed slides varying in emotional valence and arousal. The ratio of strength of dipole moments of the 2nd to the 1st P50m and the anatomical location of their sources were calculated. Negatively valenced slides significantly attenuated P50m suppression, as compared to neutral ones, while the effects of positive slides were insignificant. No effects on latencies or the location of P50m sources were observed. Thus, negative emotional stimuli may modulate sensory gating. PMID- 15812604 TI - Relationship between exploratory eye movements and brain morphology in schizophrenia spectrum patients: voxel-based morphometry of three-dimensional magnetic resonance imaging. AB - The exploratory eye movements of schizophrenia patients and their relatives have been shown to differ from those of patients without schizophrenia and healthy controls. However the mechanism of exploratory eye movement disturbances in schizophrenia patients remains elusive. We investigated the relationship between the exploratory eye movements and brain morphology in 39 schizophrenia spectrum patients. Voxel-based morphometric analysis on three-dimensional magnetic resonance imaging was conducted by means of statistical parametric mapping 99. The decrease in the responsive search score, which is the total number of sections on which the eyes fixed in response to questioning in a comparison task, was significantly correlated with the decreased gray matter in the right frontal eye field (rFEF) including the right supplementary eye field (rSEF), right parietal eye field (rPEF), and right inferior frontal region. These results suggest that disturbance in exploratory eye movement in schizophrenia spectrum patients may be related to neural network dysfunction in FEF, SEF and PEF, which are the eye movement related areas, and in the inferior frontal region that may be related to information organization. PMID- 15812605 TI - Three year course and outcome of mental illness in homeless men: a prospective longitudinal study based on a representative sample. AB - OBJECTIVE: To report on the 3-year course and outcome of mental illness and social aspects in a representative sample of 265 homeless men living on the street or using overnight shelter. METHOD: Expert interviews at baseline and at follow-up included the SCID-I and covered cognitive impairment, somatic complaints, use of medical services and psychosocial areas. Of 247 homeless men still alive, at 3-year follow-up, 185 (74.9 %) were successfully traced and interviewed face to face. RESULTS: There was considerable improvement regarding the housing situation after 3 years. Rates of mental illness decreased from 79 % to 66% over 3 years possibly due to an improved housing situation and increased medical/psychiatric attention and service. The prevalence of mood disorders, substance use disorders and anxiety disorders was significantly lower at 3-year follow-up while psychotic disorders showed a slight increase over time. Thus, in general, more remission than incidence was observed in the 3-year follow-up period. A high rate of use of general medical inpatient services was found. Considering the very high prevalence of mental illness, the use of psychiatric services was very low with some increase over time. Mental health status at 1st assessment did not predict the housing situation at 3 year follow-up. Having had a substance use disorder diagnosis at 1st assessment was a powerful predictor of an unfavorable mental health status at 3 year follow-up. Standardized mortality ratio over 3 years was 4.4. PMID- 15812606 TI - Long-term course and outcome of obsessive-compulsive patients after cognitive behavioral therapy in combination with either fluvoxamine or placebo: a 7-year follow-up of a randomized double-blind trial. AB - Longitudinal studies with very long follow-up periods of patients with obsessive compulsive disorder (OCD) who have received adequate treatment are rare. In the current study, 30 of 37 inpatients (81%) with severe OCD were followed up 6-8 years after treatment with cognitive-behavioral therapy (CBT) in combination with either fluvoxamine or placebo in a randomized design. The significant improvements (with large effectsizes) in obsessive-compulsive symptoms from pre- to post-treatment (41% reduction on the Y-BOCS) remained stable at follow-up (45 %). Responder rates, defined as > or = 35% reduction on the Y-BOCS, were 67% and 60%, respectively. Depressive symptoms decreased significantly not only from pre- to post-treatment but also during follow-up. Re-hospitalization, which occurred in 11 patients (37 %), was associated with more severe depressive symptoms at pre treatment and living without a partner. Full symptom remission at follow-up, defined as both Y-BOCS total score < or = 7 and no longer meeting diagnostic criteria for OCD, was achieved by 8 patients (27 %). Patients without full remission at follow-up had a significantly longer history of OCD, assessed at pretreatment, compared to remitted patients. The shortterm treatment outcome had no predictive value for the long-term course. Throughout the naturalistic follow up, nearly all patients (29 patients) received additional psychotherapy and/or medication. This might indicate that such chronic OCD patients usually need additional therapeutic support after effective inpatient treatment to maintain their improvements over long periods. PMID- 15812607 TI - Polymorphisms in the NMDA subunit 2B are not associated with alcohol dependence and alcohol withdrawal-induced seizures and delirium tremens. AB - OBJECTIVE: Ethanol-inhibited glutamatergic neurotransmission has been shown to mediate pathophysiological mechanisms in the development of alcoholism, including withdrawal symptoms. NMDA-receptor 2B (NR2B) is a subunit that confers a high sensitivity to ethanol-induced inhibition. Previously we had reported a lack of association between the single nucleotide polymorphism (SNP) rs1806201 in the NR2B gene (GRIN2B) and alcoholism. Shortly thereafter, an association between the polymorphism and early-onset alcoholism has been reported. One aim of the present study was to test whether the association between the GRIN2B polymorphism rs1806201 and early-onset alcoholism can be replicated in a larger sample. Moreover, we hypothesized that another genetic variation within GRIN2B (rs1806191) may have an effect in the etiology of alcoholism or withdrawal related traits. METHODS: We extended our original study sample to a size of 377 patients and 464 healthy volunteers and performed a replication study, including the second GRIN2B SNP. Associations between allele, genotype and haplotype frequencies of the two polymorphisms and alcoholism as well as with patients' phenotypes were investigated. RESULTS: No associations were found between any of the two polymorphisms, tested individually or as haplotypes, and alcoholism, respectively withdrawal-related traits. CONCLUSION: Neither the analyzed SNPs nor any of their haplotypes likely modify susceptibility to alcohol dependence or withdrawal-related phenotypes. PMID- 15812608 TI - Drug treatment modalities in psychiatric inpatient practice: a 20-year comparison. AB - OBJECTIVES: The present study investigated whether the increased number of psychotropic agents available in Italy in the last 20 years increased the proportion of inpatients being treated with individual medication classes and the proportion receiving combined treatments with agents of the same class or of different classes. METHODS: This study was conducted in South-Verona, Italy. From the local Psychiatric Case Register (PCR) all patients consecutively admitted to the inpatient unit during the years 1981/1982, 1991/92 and 2001/02 were extracted. Drug use at discharge was derived from clinical records, while service use data were extracted from the PCR. RESULTS: During the six years surveyed 160 patients were admitted in 1981/82, 139 in 1991/92 and 228 in 2001/02. An increasing proportion of subjects receiving antipsychotic, antidepressant and benzodiazepine treatment at discharge was observed. In addition, we found an increasing proportion of patients receiving two or more psychotropic drugs at discharge, which accounted for almost 80% of cases in 2001/02. The number of psychotropic agents prescribed at hospital discharge was positively correlated with the total consumption of psychotropic drugs. A relevant proportion of patients were also dispensed agents for medical conditions, yielding an average number of 3.2 prescriptions in 2001/02. The Lavik score, a summary index of service use, indicated that subjects admitted in 1981/82 were moderate users of psychiatric services, while those admitted in 1991/92 and in 2001/02 were high users of psychiatric services. CONCLUSION: This study documented emerging trends toward polypharmacotherapy and higher total doses. Additional pharmacoepidemiological research is needed to clarify both the beneficial and, potentially, adverse effects associated with these trends in psychiatric pharmacotherapy. PMID- 15812609 TI - Long-term effects of an HIV/STI sexual risk reduction intervention for heterosexual couples. AB - This randomized clinical trial examined the relative efficacy of a relationship based HIV/STI prevention intervention for women and their regular male sexual partners at 12 months post-intervention. A total of 217 couples were randomized to (1) a six-session intervention provided to the woman and her sexual partner together (n=81); (2) the same intervention provided to the woman alone (n=73); or (3) a one-session health information education "control" provided to the woman alone (n=63). Findings suggest the intervention was efficacious in reducing unprotected sex at 12 months post-intervention, compared with the education control group. No significant differences were observed when comparing whether couples received the intervention together or when the woman received it alone. PMID- 15812610 TI - Disclosure of HIV status and psychological well-being among Latino gay and bisexual men. AB - This study examined disclosure of HIV-positive serostatus by 301 Latino gay and bisexual men to members of their social networks and the mental health consequences of such disclosure. The sample was recruited from clinics, hospitals, and community agencies in New York City, Washington, DC, and Boston. Proportions disclosing differed depending on the target, with 85% having disclosed to closest friend, 78% to male main partner, 37% to mother, and 23% to father. Although there were differences depending on the target, disclosure was related to greater quality of social support, greater self-esteem, and lower levels of depression. Moreover, findings indicated that social support mediated the relationship between disclosure of serostatus and both self-esteem and depression. Thus, disclosure resulted in greater social support, which in turn had positive effects on psychological well-being. Findings demonstrate that generally Latino gay men are selective in choosing people to whom they disclose their serostatus and that disclosure tends to be associated with positive outcomes. PMID- 15812611 TI - Drug sharing among heroin networks: implications for HIV and hepatitis B and C prevention. AB - Qualitative and quantitative findings from the baseline survey of a longitudinal, socially-focused blood-borne disease intervention study among 611 heroin IDU in Denver indicate that high risk injection practices-the sharing of contaminated drug solution in particular-often occur as a consequence of how heroin is obtained, the quantity obtained and the setting where it is injected. Contamination occurs if a contaminated syringe is used to liquefy and apportion the shared drug. In our cohort of 304 heroin injecting networks there was at least one member who, when asked to describe their last injection, reported dividing the drug as a liquid (82%), using a reservoir of water that syringes had been rinsed in to mix drugs (67%), using a common cooker (86%)-a proxy for drug sharing-and beating a shared cotton filter (58%). In contrast, only 22% reported syringe sharing. Variables associated with various injection practices included location of the last injection episode, quantity of drug injected, dope sickness, and years injecting. When compared to those who injected in a safe setting, those in an unsafe location had almost three times the odds (OR = 2.9; 95% CI: 1.9, 4.6) of being part of an injection episode where there was cooker sharing; and the smaller the quantity of heroin (< or =1/4 gram v. > 1/4 gram) present at the episode, the greater the odds that cooker sharing occurred (OR = 1.8; 95% CI: 1.2, 2.6). Use of a used, unbleached syringe to prepare shared drugs had twice the odds of occurring in "unsafe" v. safe settings (OR = 2.2; 95% CI: 1.3, 4.0) and in episodes in which a participant was dopesick (OR = 2.1; 95% CI: 1.2, 3.6). In summary, risky injection practices occur within an injection process that is, in part, a response to a structurally imposed risk environment. Lessening the blood-borne disease risks embedded within this process requires interventions designed to mitigate the environmental factors that influence it, including syringe accessibility, law enforcement strategies and the settings where IDU inject drugs. PMID- 15812612 TI - Factors associated with HIV testing, condom use, and sexually transmitted infections among female sex workers in Nha Trang, Vietnam. AB - This study examined predictors of HIV testing, successful condom negotiation with clients, and self-reported sexually transmitted infections (STIs) among Vietnamese female sex workers (FSW). Data were collected by using face-to-face interviews from a community sample of 610 FSW from Nha Trang city during October December, 2000. Having had an HIV test was associated with having spent time in a rehabilitation center. Consistently successful negotiation of condom use occurred most among FSW who had few clients, understood how HIV was not transmitted, and had not reported ever having any symptoms of STIs. Migration to Nha Trang for sex work was a risk factor for an STI diagnosis; successful negotiation of condom use had a protective effect. Our results suggest the need for voluntary HIV counseling and testing, further promotion of condom use among FSW populations, and better use of rehabilitation sites to promote HIV prevention. PMID- 15812613 TI - Risk factors for HIV-AIDS among youth in Cape Town, South Africa. AB - South Africa is in the midst of a devastating HIV-AIDS epidemic and most new HIV infections occur among young adults and adolescents. The current study examined risk behaviors and HIV risk factors among young people living in a Black South African township. Using community-based outreach methods of street intercept and facility-based surveying, 113 men and 115 women age 25 and younger responded to an anonymous survey. Results showed that men (68%) and women (56%) reported HIV related high risk sexual behaviors. Although knowledge about HIV transmission was generally high, there was evidence that misconceptions about AIDS persist, particularly myths related to HIV transmission. For young men, HIV risk factors were associated with fewer years of education, lower levels of AIDS-related knowledge, condom attitudes, and Dagga (marijuana) use. Among young women, HIV risk factors were associated with beliefs that condoms get in the way of sex and rates of unprotected vaginal intercourse. Despite adequate general AIDS knowledge and risk sensitization, South African youth demonstrated high rates of sexual practices that place them at risk for HIV infection. There is an urgent need for behavioral interventions targeted to young South Africans living in the most economically disadvantaged areas. PMID- 15812614 TI - Post-circuit blues: motivations and consequences of crystal meth use among gay men in Miami. AB - Miami, Florida was at the vanguard of the rise of circuit parties and attendant club drug use-especially ecstasy, GHB, and ketamine-in the 1990s. Crystal methamphetamine, a drug of abuse among gay men for some years on the West coast, gradually moved east toward the end of the decade and recently became prevalent in Miami. This paper reports the results of focus group research into the motivations and consequences of crystal use among gay men in this new setting. Loneliness, fears about physical attractiveness due to aging and illness, and desires to lose sexual inhibitions were common motivations for using the drug. Continued use of crystal was often described as the cause of lost friendships, employment and long-term relationships, as well as sexual behaviors that put men at risk for HIV and other sexually transmitted infections. Implications for drug and sexual risk prevention interventions are discussed. PMID- 15812615 TI - Isolating the nexus of substance use, violence and sexual risk for HIV infection among young adults in the United States. AB - Adults aged 18 to 29 are at significant sexual risk for HIV infection. Substance use and violence are known to be associated with sexual risk in certain groups, but few studies have examined these relationships in the general population of young adults. No studies have tested whether the contributions of substance use and violence to sexual risk are independent, and few have looked at whether drug use associations with risk are specific to certain substances. Using structural modeling techniques, we examined data for 3,437 adults aged 23-24, testing for associations between three measures of sexual risk for HIV, various forms of substance use, victimization and partner violence. Alcohol use and victimization predicted high risk sex in independent samples of single and married/cohabiting adults. Marijuana use, problem drug use, and partner violence were inconsistently related to sexual risk across measures and subsamples. HIV-prevention interventions designed for young adults in the general population should target individuals who use alcohol frequently and who are victims of violence, and should address both factors, in addition to sexual risk behavior. PMID- 15812617 TI - Use of electronic monitoring devices to measure antiretroviral adherence: practical considerations. AB - The purpose of this paper is to describe electronic monitoring device (EMD) (e.g., MEMS caps) use among HIV-infected adults enrolled in a randomized clinical trial and to make explicit some of the benefits and caveats of using electronic monitoring device technology. This is a descriptive, exploratory study of EMD use among 128 HIV-infected adults treated with at least three antiretroviral agents. Thirty-six percent of the sample admitted that they did not use the EMD consistently. Forty-one percent of the subjects reported taking out more than one dose at a time and 26% reported opening the EMD but not taking the medication. Special subject-related issues accounted for only a small percentage of all reported problems with EMD use (e.g., transient housing, incarceration, substance abuse relapse and drug treatment). Results of this study suggest that EMDs may underestimate antiretroviral adherence among HIV-infected adults. Recommendations for improving EMD data quality are presented. PMID- 15812616 TI - Patient-clinician relationships and treatment system effects on HIV medication adherence. AB - The study objectives were to determine the impact of the patient-clinician relationship on patient adherence to HIV medication, to identify which aspects of the patient-clinician relationship and the treatment system influenced adherence, and to determine which of these variables remained important when the impact of mental distress and substance abuse were considered. The design was a cross sectional study using a sample of 120 HIV+ clinic patients. The Primary Care Assessment Survey (PCAS) assessed the clinician-patient relationship and the treatment system. The Composite International Diagnostic Inventory-Short Form (CIDI-SF) screened for mental disorders, and the Brief Substance Abuse History Form measured recent and remote substance use. Patient adherence was assessed using five markers including 3 interview-elicited self-reports, 1 medical chart review, and 1 summary score. Logistic regression analyses identified independent predictors of each adherence behavior. PCAS scores contributed to all five models, and their effects persisted when mental distress and substance abuse were considered. Adherence behaviors are explained by a variety of factors and should be assessed using multiple methods. Further study to illuminate the mechanisms of action of the clinician-patient relationship on adherence to HIV medication is warranted. PMID- 15812618 TI - Demographic characteristics, treatment history, drug risk behaviors, and condom use attitudes for U.S. and Russian injection drug users: the need for targeted sexual risk behavior interventions. AB - Two separate databases, one on Russian (n = 444) injection drug users (IDUs), and the other on U.S. IDUs (n = 241), were merged, and responses were compared. Results indicated that Russian IDUs perceived themselves to be at greater risk for HIV/AIDS based upon behaviors over the past 90 days. U.S. IDUs were more likely to be tested for HIV, report a negative HIV result, and know more people with HIV. U.S. IDUs consumed greater amounts of alcohol and marijuana, and made more alcohol/drug treatment attempts than Russian IDUs. Russian IDUs injected more frequently, although were more likely to recently use clean needles. Russian and U.S. IDUs responded significantly differently to eight AIDS knowledge questions. Eight condom attitude questions were asked of each group, and each revealed significantly different responses. Gender differences for Russian IDUs were also observed. There is a clear need to create culturally targeted sexual risk reduction interventions. PMID- 15812619 TI - A framework for examining the sustained acceptability of microbicides. AB - The concept of acceptability involves a complex set of individual, couple and socio-cultural factors. Within a specific focus on microbicides, this article sets forth the relevant behavioral issues demanding attention as scientists and product developers strive to identify microbicides that will be used in a sustained, long-term fashion to prevent HIV/AIDS. The paper re-conceptualizes long-term acceptability within the ARRM framework, reviews the basic issues and research related to microbicide acceptability, discusses couple relationships (especially the nature of intimate relationships), and identifies related gender and power factors. The importance of the cultural context within various international settings pervades all of these issues. Finally, a set of challenges is discussed for those contemplating such research. PMID- 15812620 TI - [Simulation and airway management]. AB - Despite the development of new devices and strategies to manage and secure the difficult airway, morbidity and mortality in anaesthesia due to airway problems such as difficult intubation or unrecognised failed intubation remain high. The problem seems to lie in the transfer of skills and strategies to daily clinical practice. Common methods for airway management training include theoretical instructions and hands-on sessions with manikins, animal models and cadavers. Simulation provides the opportunity to train skills and resolve specific situations embedded in a realistic scenario, facilitate the transfer of cognitive, psychomotor and affective abilities into daily clinical practice and help to improve behaviour in critical situations. This article outlines new training concepts in airway management with the help of simulation and simulators. We describe technical prerequisites and provide information on the implementation of difficult airway scenarios. PMID- 15812621 TI - [The influence of the C-leg knee-shin system from the Otto Bock Company in the care of above-knee amputees. A clinical-biomechanical study to define indications]. AB - The C-Leg microprocessor-controlled knee-shin system for the above-knee amputees is introduced as a dramatic improvement over all other prosthetic knees. This is due to its combination of on-board microprocessor and the hydraulic controls acting both on the swing and stance phase. A more secure, natural and efficient gait is expected. Following the recommendations of Otto Bock the indications for the prescription of the C-leg are: Amputees with mobility level "able to walk outdoors without limitations" (AK3) and "able to walk outdoors without limitations plus engage in high performance activities" (AK4) if they face at least one extra obstacle as listed in the Otto Bock catalogue of indications. In this article it is aimed to critically review the indications for the C-leg. In particular the question is posed, whether a different or sophisticated indication of mobility levels might be suggested. Therefore this study does not concentrate on the 3C-100 C-Leg((R)) component but on the system patient + C-leg. So the testing is done by comparing the C-Leg against the regular knee, which is assumed to be an adequate choice for this patient and to which he is accustomed. So far 25 patients with activity levels AK 2 (5), AK 3 (13) and AK 4 (7) have participated in the study. 23 patients, i.e. all patients except one AK 2 and one AK 3 exhibit functional improvement at least according to one criterion. On the other side, only three patients (2 AK 4), fulfill all criteria of functional improvement, which have been defined for this test. It is concluded, that multi handicapped patients of all activity levels generally experience substantial improvement due to this system. AK 2 patients may show significant functional improvement. As a prerequisite, however, they must not exhibit deficiencies regarding stump movement, muscular status or cognitive abilities. Active patients (AK 3 and AK 4) benefit in the majority of cases. However, some highly active patients of AK 4 complain about interferences between their intended movement and the microprocessor control of knee movements. PMID- 15812622 TI - The concept of "baby lung". AB - BACKGROUND: The "baby lung" concept originated as an offspring of computed tomography examinations which showed in most patients with acute lung injury/acute respiratory distress syndrome that the normally aerated tissue has the dimensions of the lung of a 5- to 6-year-old child (300-500 g aerated tissue). DISCUSSION: The respiratory system compliance is linearly related to the "baby lung" dimensions, suggesting that the acute respiratory distress syndrome lung is not "stiff" but instead small, with nearly normal intrinsic elasticity. Initially we taught that the "baby lung" is a distinct anatomical structure, in the nondependent lung regions. However, the density redistribution in prone position shows that the "baby lung" is a functional and not an anatomical concept. This provides a rational for "gentle lung treatment" and a background to explain concepts such as baro- and volutrauma. CONCLUSIONS: From a physiological perspective the "baby lung" helps to understand ventilator-induced lung injury. In this context, what appears dangerous is not the V(T)/kg ratio but instead the V(T)/"baby lung" ratio. The practical message is straightforward: the smaller the "baby lung," the greater is the potential for unsafe mechanical ventilation. PMID- 15812623 TI - Courts, doctors and end-of-life care. PMID- 15812624 TI - Effects of levosimendan on systemic and regional hemodynamics in septic myocardial depression. AB - OBJECTIVE: Calcium desensitization plays an important part in the pathophysiology of septic myocardial depression. We postulated that levosimendan, a new calcium sensitizer, would be beneficial in sepsis-induced cardiac dysfunction. DESIGN AND SETTING: Prospective, randomized, controlled study in two university hospital intensive care units. PATIENTS AND PARTICIPANTS: Twenty-eight patients with persisting left ventricular dysfunction related to septic shock after 48 h of conventional treatment including dobutamine (5 microg/kg per minute). INTERVENTIONS: After 48 h of conventional treatment patients were randomized to receive a 24-h infusion of either levosimendan (0.2 microg/kg per minute, n=15) or dobutamine (5 microg/kg per minute, n=13). MEASUREMENTS AND RESULTS: Data from right heart catheterization, echocardiography, gastric tonometry, laser-Doppler flowmetry, and lactate concentrations and creatinine clearance were obtained before and after the 24-h drug infusion. Dobutamine did not change systemic or regional hemodynamic variables. By contrast, at the same mean arterial pressure levosimendan decreased pulmonary artery occlusion pressure and increased cardiac index. Levosimendan decreased left ventricular end-diastolic volume and increased left ventricular ejection fraction. Levosimendan increased gastric mucosal flow, creatinine clearance, and urinary output while it decreased lactate concentrations. CONCLUSIONS: These findings show that levosimendan improves systemic hemodynamics and regional perfusion in patients with septic cardiac dysfunction under conditions where administration of 5 microg/kg dobutamine per minute is no longer efficacious. Accordingly, our results suggest that levosimendan can be an alternative to the strategy of increasing the dose of dobutamine under such conditions. PMID- 15812625 TI - Blockade of nuclear factor-kappaB activation prevents hypodynamic shock and gastric hypoperfusion induced by endotoxin in anesthetized dogs. AB - OBJECTIVE: To investigate whether pyrrolidinone derivative (N2733), an inhibitor of nuclear factor (NF)-kappaB activation, improves altered metabolic and hemodynamic changes and organ dysfunctions caused by endotoxic shock. DESIGN AND SETTING: Prospective, randomized, animal study in a laboratory at a university hospital. SUBJECTS: Twenty-three anesthetized male beagle dogs (10-14 kg). INTERVENTIONS: Dogs were mechanically ventilated and monitored with a pulmonary arterial catheter and a gastric tonometer. A central venous catheter was inserted into the femoral vein, and lactated Ringer's solution (10 ml/kg per hour) was administered throughout the study period. Three groups of animals were studied: (a) the lipopolysaccharide (LPS) group (n=8), which received LPS (250 ng/kg per minute for 2 h); (b) the LPS plus N2733 group (n=8), which received N2733 (30 mg/kg intravenously and 10 mg/kg hour for 6 h) after the start of LPS; and (c) the N2733 group (n=7), which received N2733 (30 mg/kg intravenously and 10 mg/kg per hour for 6 h). MEASUREMENTS AND RESULTS: Changes in hemodynamics, blood gas, gastric intramural pH, and renal and hepatic function were measured for 6 h. Coadministration of N2733 increased oxygen delivery index and prevented the LPS induced hypotension, metabolic acidosis, and gastric mucosal acidosis but did not affect renal or hepatic function. CONCLUSIONS: Administration of N2733 increased oxygen delivery index and prevented the LPS-induced hypotension and metabolic and gastric mucosal acidosis in an anesthetized canine endotoxic shock model, suggesting its beneficial effect on local blood flow against tissue hypoxia. These findings suggest that blockade of NF-kappaB activation prevents hypodynamic shock and gastric hypoperfusin in endotoxic shock. PMID- 15812626 TI - Weaning failure from mechanical ventilation due to hypertrophic obstructive cardiomyopathy. AB - OBJECTIVE: To report the importance of hypertrophic obstructive cardiomyopathy as a potential substrate for difficult weaning from mechanical ventilation. DESIGN AND SETTING: Case report in an adult intensive care unit of a tertiary care hospital. PATIENTS: Two patients who developed intractable pulmonary edema with subsequent difficult weaning from mechanical ventilation in the early postoperative period despite medical treatment for heart failure based on pulmonary artery catheter data. INTERVENTIONS: Mechanical ventilation, medical treatment. MEASUREMENTS AND RESULTS: Pulmonary artery catheter data in these two elderly patients under mechanical ventilation were compatible with congestive heart failure, and the initial therapy was diuretics, vasodilators, and inotropes. Despite this reasonable medical treatment no hemodynamic improvement and thereby failure of weaning from mechanical ventilation was observed. Transesophageal echocardiography demonstrated hypertrophic obstructive cardiomyopathy in both cases, and therapy based on negative inotropes was instituted. This treatment change induced hemodynamic improvement and successful weaning from mechanical ventilation. CONCLUSIONS: In mechanically ventilated patients because of congestive heart failure who have difficult weaning, despite appropriate therapy, intensivists should always suspect causes other than the obvious systolic heart failure. In this clinical setting echocardiography is warranted and produces excellent diagnostic yield for clarifying complex cardiac disturbances. PMID- 15812627 TI - Beneficial hemodynamic response of transthoracic cardiac pacing in a 2 kg preterm neonate. AB - OBJECTIVE: To describe the beneficial hemodynamic effect of transthoracic external pacing compared with epicardial pacing in a 2 kg premature neonate with congenital complete heart block, hydrops and complex congenital heart disease. METHODS: Transthoracic epicardial pacing was instituted at a rate of 120 beats/min to treat life-threatening bradycardia (HR 50-60 beats/min), hypotension (BP 45/20 mmHg) and severe lactic acidosis (pH 7.18, lactate 7.5 mmol/l) despite significant catecholamine support. Due to the size of the patient, the external pacing electrodes were placed on the back and front of the patient's chest. To achieve ventricular capture, the electric output had to be increased to 140 mA, resulting in symmetric chest movements. RESULT: Transthoracic pacing resulted in an immediate improvement of the cardiovascular status, with an adequate blood pressure (BP 60/25 mmHg) and normalization of acid-base-balance. After the insertion of an epicardial pacemaker (paced rate at 140 beats/min), a significantly lower blood pressure was achieved (BP 50/25 mmHg), indicating an additive effect of the rhythmic chest movements to the blood pressure. Unfortunately, third-degree burns were detected on the patients back 1 day after pacemaker insertion. CONCLUSION: Transthoracic pacing is a life-saving option in an emergency situation, when maximal pharmacological support fails to maintain adequate cardiac output. It may have some additional beneficial effect on blood pressure generation in these patients. To prevent skin injury, the output used needs to be as low as feasible, and the period of pacing as brief as practicable. PMID- 15812628 TI - Small increases in the urinary excretion of glutathione S-transferase A1 and P1 after cardiac surgery are not associated with clinically relevant renal injury. AB - OBJECTIVE: Cardiac surgery is an important risk factor for the development of acute renal failure. Cytosolic enzymes glutathione S-transferase (GST) A1 and P1 are present selectively in proximal and distal tubular cells, respectively. We determined the extent and site of tubular injury and examined if GST excretion may predict a clinically relevant change in renal function. DESIGN AND SETTING: A prospective, observational study in 84 consecutive patients in the cardiac surgery intensive care unit of the University Medical Centre Nijmegen. MEASUREMENTS AND RESULTS: Urinary GST enzyme excretion was determined 0-4 h and 20-24 h after cardiac surgery by enzyme-linked immunosorbent assay. Data are expressed as median and 5-95% range. Urinary excretion of GSTA1 was increased: 1.25 microg/mmol [0.31-10.20] creatinine at t =0-4 h ( p <0.0001, compared with controls; 0.25 [0.1-0.8]) and returned to normal values at t =20-24 h. Excretion of GSTP1 was 2.11 microg/mmol [0.52-17.82] creatinine ( p <0.0001) at t =0-4 h and remained significantly elevated: 0.84 [0.30-16.86] at t =20-24 h ( p =0.01) compared with controls (0.5 [0.2-1.1]). The ten patients with the highest urinary excretion of GSTA1 or GSTP1 did not demonstrate a different plasma creatinine level on postoperative day 3, compared with the ten patients with the lowest urinary excretion of GSTA1 or GSTP1. CONCLUSION: Uncomplicated cardiac surgery results in a statistically significant increase in the urinary excretion of GSTA1 and GSTP1 as compared with healthy controls, indicating proximal and distal tubular damage. However, this small increase in urinary excretion of GSTs is not associated with clinically relevant renal injury. PMID- 15812630 TI - Cytoplasmic contractions in growing fungal hyphae and their morphogenetic consequences. AB - Video-enhanced light microscopy of the apical and subapical regions of growing hyphae of several fungal species revealed the existence of momentary synchronized motions of subcellular organelles. First discovered in a temperature-sensitive morphological mutant (ramosa-1) of Aspergillus niger, these seemingly spontaneous cytoplasmic contractions were also detected in wild-type hyphae of A. niger, Neurospora crassa, and Trichoderma atroviride. Cytoplasmic contractions in all fungi lasted about 1 s. Although the cytoplasm recovered its motility and appearance, the contraction usually led to drastic changes in Spitzenkorper (apical body) behavior and hyphal morphology, often both. Within 10 s after the contraction, the Spitzenkorper commonly became dislodged from its polar position; sometimes it disassembled into phase-dark and phase-light components; more commonly, it disappeared completely. Whether partial or complete, the dislocation of the Spitzenkorper was always accompanied by a sharp reduction or cessation of growth, and was usually followed by marked morphological changes that included bulbous hyphal tips, bulges in the hyphal profile, and formation of subapical and apical branches. The cytoplasmic contractions are vivid evidence that the most conspicuous cell organelles (membrane-bound) in living hyphae are interconnected via a contractile cytoskeletal network. PMID- 15812631 TI - Overexpression, purification and characterization of SimL, an amide synthetase involved in simocyclinone biosynthesis. AB - Simocyclinone D8 is a potent inhibitor of bacterial gyrase, produced by Streptomyces antibioticus Tu 6040. It contains an aminocoumarin moiety, similar to that of novobiocin, which is linked by an amide bond to a structurally complex acyl moiety, consisting of an aromatic angucycline polyketide nucleus, the deoxysugar olivose and a tetraene dicarboxylic acid. We have now investigated the enzyme SimL, responsible for the formation of the amide bond of simocyclinone. The gene was cloned, expressed in S. lividans T7, and the protein was purified to near homogeneity, and characterized. The 60 kDa protein catalyzed both the ATP dependent activation of the acyl component as well as its transfer to the amino group of the aminocoumarin ring, with no requirement for a 4'-phosphopantetheinyl cofactor. Besides its natural substrate, simocyclinone C4, SimL also accepted a range of cinnamic and benzoic acid derivatives and several other, structurally very diverse acids. These findings make SimL a possible tool for the creation of new aminocoumarin antibiotics. PMID- 15812629 TI - Effects of volumetric vs. pressure-guided fluid therapy on postoperative inflammatory response: a prospective, randomized clinical trial. AB - OBJECTIVE: To compare intrathoracic blood volume (ITBV) guided fluid management and central venous pressure (CVP) guided therapy in ameliorating the progression of early systemic inflammatory response in patients undergoing major surgery. DESIGN: Prospective, randomized clinical trial. PATIENTS: Forty patients undergoing major abdominal surgery were randomized into CVP and ITBV groups. INTERVENTIONS: In the CVP group the target CVP was 8-12 mmHg while in the ITBV group the goal was to keep the ITBV between 850 and 950 ml/m2 during the operation. MEASUREMENTS AND RESULTS: Hemodynamic parameters were determined by single arterial thermodilution. Measurements were repeated every 30 min intraoperatively. Serum procalcitonin (PCT) and C-reactive protein (CRP) was monitored preoperatively, on ICU admission, and then daily for 3 days. Serum TNF alpha levels were measured intraoperatively hourly and then daily for 3 days. There was no significant difference between the two groups regarding hemodynamic parameters at any assessment point. In the overall population changes in the stroke volume index showed a significant correlation with changes in CVP and ITBV. TNF-alpha levels remained in the normal range intraoperatively and during the three postoperative days in both groups. Preoperatively normal PCT and CRP levels increased significantly postoperatively, without significant differences between the groups. CONCLUSIONS: ITBV guided fluid therapy did not alter the magnitude of inflammatory response as monitored by serum PCT, CRP, and TNF-alpha in the early postoperative period. PMID- 15812632 TI - Ehlers-Danlos syndrome type IV and recurrent carotid-cavernous fistula: review of the literature, endovascular approach, technique and difficulties. AB - We report the follow-up of a previously published case (Forlodou et al. Neuroradiology 38:595-597, 1996) of carotido-cavernous fistulas (CCFs) in a patient presenting with type IV Ehlers-Danlos syndrome (EDS 4) that were successfully treated twice by an endovascular approach. Initial treatment with a detachable balloon was in 1994 for a right CCF, and, 8 years later, a left CCF was treated by selective transarterial occlusion of the cavernous sinus with coils. Unfortunately, the patient suffered from a spontaneous post-operative intracranial haemorrhage in the left hemisphere and died. Review of the literature, technical considerations for bilateral CCF and complication are discussed. PMID- 15812633 TI - Fifteen-year follow-up of a patient with beta thalassaemia and extramedullary haematopoietic tissue compressing the spinal cord. AB - A long-term follow-up of a patient with beta thalassaemia with intra- and extraspinal extramedullary haematopoietic tissue compressing the spinal cord is presented. Extramedullary haematopoietic nodules are a rare cause of spinal cord compression and should be included in the differential diagnosis, especially in patients from Mediterranean countries. Treatment with radiation therapy solely failed, giving rise to the need of surgical intervention. Surgical decompression of the spine and the removal of the culprit lesion compressing the spine were performed. Postinterventional radiation therapy was applied to the spine. A relapse had to be treated again by surgical means combined with postinterventional radiation therapy. A complete relief of the symptoms and control of the lesion could be obtained. PMID- 15812634 TI - Pontomedullary disconnection: fetal and neonatal considerations. AB - The cerebellar and pontocerebellar hypoplasias present a unique challenge when detected in the developing fetus. A diverse aetiology and prognosis make counselling of these families difficult. Advances in fetal imaging allow for more accurate diagnosis and counselling, but postnatal MRI is still required. A case is presented in which cerebellar hypoplasia was detected at 20 weeks gestation. Later fetal imaging provided further information, but a diagnosis of pontomedullary disconnection was not made until the postnatal MRI scan. The clinical findings and possible causes of such pontocerebellar abnormalities are discussed. PMID- 15812635 TI - Sunburst periosteal reaction in osteogenic sarcoma. PMID- 15812636 TI - The role of the periplasmic loop residue glutamine 65 for MscL mechanosensitivity. AB - The periplasmic loop of MscL, the mechanosensitive channel of large conductance, acts as a spring resisting the opening of the channel. Recently, a high throughput functional screening of a range of MscL structural mutants indicated that the substitution of residue glutamine (Q) 65 with arginine (R) or leucine (L) leads to a wild-type (WT)-like and a loss-of-function (LOF) phenotype, respectively. We used electron paramagnetic resonance (EPR) spectroscopy, single channel recording and in vivo experiments to investigate further the effect of R and L mutation of Q65 on the gating mechanism of MscL. Structural analysis of Q65R and Q65L was carried out by coupling the site-directed spin labeling (SDSL) with EPR spectroscopy. A SDSL cysteine mutant of the isoleucine 24 residue (I24C SL) in the first transmembrane domain, TM1, of MscL served as a reporter residue in EPR experiments. This was due to its strong spin-spin interaction with the neighboring I24C-SL residues in the MscL channel pentamer. The effects of bilayer incorporation of lysophosphatidylcholine on the MscL mutants were also investigated. Functional analysis was carried out using patch-clamp recordings from these mutants and WT MscL reconstituted into artificial liposomes. Although our data are largely in agreement with the high-throughput mutational analysis of Maurer and Dougherty, this study shows that Q65R and Q65L form functional channels and that these mutations lead to partial gain-of-function (GOF) and LOF mutation, respectively. Overall, our study confirms and advances the notion that the periplasmic loop plays a role in setting the channel mechanosensitivity. PMID- 15812637 TI - Visualisation of the mechanosensitive channel of large conductance in bacteria using confocal microscopy. AB - The mechanosensitive channel of large conductance (MscL) plays an important role in the survival of bacterial cells to hypo-osmotic shock. This channel has been extensively studied and its sequence, structure and electrophysiological characteristics are well known. Here we present a method to visualise MscL in living bacteria using confocal microscopy. By creating a gene fusion between mscl and the gene encoding the green fluorescent protein (GFP) we were able to express the fusion protein MscL-GFP in bacteria. We show that MscL-GFP is present in the cytoplasmic membrane and forms functional channels. These channels have the same characteristics as wild-type MscL, except that they require more pressure to open. This method could prove an interesting, non-invasive, tool to study the localisation and the regulation of expression of MscL in bacteria. PMID- 15812638 TI - Refined NMR structure of alpha-sarcin by 15N-1H residual dipolar couplings. AB - (15)N-(1)H residual dipolar couplings (RDC) have been used as additional restraints to refine the solution structure of the ribotoxin alpha-sarcin. The RDC values were obtained by partial alignment of alpha-sarcin in the binary mixture of n-dodecyl hexa(ethylene glycol)/hexanol. A total of 131 RDCs were measured and 106 were introduced in the final steps of the calculation protocol following the main calculation based on nuclear Overhauser enhancements and torsion angle restraints. A homogeneous family of 81 conformers was obtained. The resulting average pairwise root-mean-square deviation corresponding to the superposition of the 20 best structures is 0.69+/-0.12 A for the backbone and 1.29+/-0.14 A for all heavy atoms. The new structural features derived from the refined structure, compared with the non-refined structure of alpha-sarcin, consist of new hydrogen bonds and a better definition of the backbone conformation. In particular, the loop segment spanning Gly 60 to Lys 70 shows a single conformation, corresponding to the most populated family of conformers observed in the unrefined structure. The information derived from the analysis of the refined structure and the comparison with the homologous protein restrictocin could help in establishing further structure-function relationships concerning alpha-sarcin which can be reasonably extrapolated to other members of the ribotoxin family. PMID- 15812639 TI - Ligand rebinding: self-consistent mean-field theory and numerical simulations applied to surface plasmon resonance studies. AB - Rebinding of dissociated ligands from cell surface proteins can confound quantitative measurements of dissociation rates important for characterizing the affinity of binding interactions. This can be true also for in vitro techniques such as surface plasmon resonance (SPR). We present experimental results using SPR for the interaction of insulin-like growth factor-I (IGF-I) with one of its binding proteins, IGF binding protein-3 (IGFBP-3), and show that the dissociation, even with the addition of soluble heparin in the dissociation phase, does not exhibit the expected exponential decay characteristic of a 1:1 binding reaction. We thus consider the effect of (multiple) rebinding events and, within a self-consistent mean-field approximation, we derive the complete mathematical form for the fraction of bound ligands as a function of time. We show that, except for very low association rate and surface coverage, this function is nonexponential at all times, indicating that multiple rebinding events strongly influence dissociation even at early times. We compare the mean field results with numerical simulations and find good agreement, although deviations are measurable in certain cases. Our analysis of the IGF-I-IGFBP-3 data indicates that rebinding is prominent for this system and that the theoretical predictions fit the experimental data well. Our results provide a means for analyzing SPR biosensor data where rebinding is problematic and a methodology to do so is presented. PMID- 15812640 TI - Alignment of biological microparticles by a polarized laser beam. AB - The optical alignment of biological samples is of great relevance to microspectrometry and to the micromanipulation of single particles. Recently, Bayoudh et al. (J. Mod. Opt. 50:1581-1590, 2003) have shown that isolated, disk shaped chloroplasts can be aligned in a controlled manner using an in-plane polarized Gaussian beam trap, and suggested that this is due to their nonspherical shape. Here we demonstrate that the orientation of various micrometer-sized isolated biological particles, trapped by optical tweezers, can be altered in a controlled way by changing the plane of linear polarization of the tweezers. In addition to chloroplasts, we show that subchloroplast particles of small size and irregular overall shape, aggregated photosynthetic light harvesting protein complexes as well as chromosomes can be oriented with the linearly polarized beam of the tweezers. By using a laser scanning confocal microscope equipped with a differential polarization attachment, we also measured the birefringence of magnetically oriented granal chloroplasts, and found that they exhibit strong birefringence with large local variations, which appears to originate from stacked membranes. The size and sign of the birefringence are such that the resulting anisotropic interaction with the linearly polarized laser beam significantly contributes to the torque orienting the chloroplasts. PMID- 15812641 TI - Sugar uptake and sensitivity to carbon catabolite regulation in Streptomyces peucetius var. caesius. AB - Streptomyces peucetius var. caesius produces a family of secondary metabolites called anthracyclines. Production of these compounds is negatively affected in the presence of glucose, galactose, and lactose, but the greatest effect is observed under conditions of excess glucose. Other carbon sources, such as arabinose or glutamate, show either no effect or stimulate production. Among the carbon sources that negatively affect anthracycline production, glucose is consumed in greater concentrations. We determined glucose and galactose transport in S. peucetius var. caesius and in a mutant of this strain whose anthracycline production is insensitive to carbon catabolite repression (CCR). In the original strain, incorporation of glucose and galactose was stimulated when the microorganism was grown in media containing these sugars, although we also observed basal galactose incorporation. Both the induced and the basal incorporation of galactose were suppressed when the microorganism was grown in the presence of glucose. Furthermore, adding glucose directly during the transport assay also inhibited galactose incorporation. In the mutant strain, we observed a reduction in both glucose (48%) and galactose (81%) incorporation compared to the original. Galactose transport in this mutant showed reduced sensitivity to the negative effect of glucose; however, it was still sensitive to inhibition. The deficient transport of these sugars, as well as CCR sensitivity to glucose in this mutant was corrected when the mutant was transformed with the SCO2127 region of the Streptomyces coelicolor genome. Our results support a role for glucose as the most easily utilized carbon source capable of exerting the greatest repression on anthracycline biosynthesis. In consequence, glucose also prevented the repressive effect of galactose by suppressing its incorporation. This suggests the participation of an integral regulatory system, which is initiated by an increase in incorporation of repressive sugars and their metabolism as a prerequisite for establishing the phenomenon of CCR in S. peucetius var. caesius. PMID- 15812642 TI - Characterization of Sphingomonas aldehyde dehydrogenase catalyzing the conversion of various aromatic aldehydes to their carboxylic acids. AB - An aldehyde dehydrogenase gene, designated phnN, was isolated from a genome library of the 1,4-dimethylnaphthalene-utilizing soil bacterium, Sphingomonas sp. 14DN61. Escherichia coli expressing the phnN gene converted 1,4 dihydroxymethylnaphthalene to 1-hydroxymethyl-4-naphthoic acid. The putative amino acid sequence of the phnN gene product had 31-42% identity with those of NAD(+)-dependent short-chain aliphatic aldehyde dehydrogenases and a secondary alcohol dehydrogenase. The NAD(P)(+)-binding site and two consensus sequences involved in the active site for aldehyde dehydrogenase are conserved among these proteins. The PhnN enzyme purified from recombinant E. coli showed broad substrate specificity towards various aromatic aldehydes, i.e., 1- and 2 naphaldehydes, cinnamaldehyde, vanillin, syringaldehyde, benzaldehyde and benzaldehydes substituted with a hydroxyl, methyl, methoxy, chloro, fluoro, or nitro group were converted to their corresponding carboxylic acids. Interestingly, E. coli expressing phnN was able to biotransform a variety of not only aromatic aldehydes, but also aromatic alcohols to carboxylic acids. PMID- 15812643 TI - Rat cytochrome P450-mediated transformation of dichlorodibenzo-p-dioxins by recombinant white-rot basidiomycete Coriolus hirsutus. AB - Rat cytochrome P450, CYP1A1, has been reported to play an important role in the metabolism of mono-trichlorodibenzo-p-dioxins (M-TriCDDs). To breed lignin (and M TetraCDDs)-degrading basidiomycete Coriolus hirsutus strains producing rat CYP1A1, an expression cassette [C. hirsutus gpd promoter-C. hirsutus gpd 5' portion (224-bp of 1st exon-8th base of 4th exon)-rat cyp1a1 cDNA-Lentinula edodes priA terminator] was constructed and inserted into pUCR1 carrying the C. hirsutus arg1 gene. The resulting recombinant plasmid, MIp5-(cyp1a1 + arg1) was introduced into protoplasts of C. hirsutus monokaryotic strain OJ1078 (Arg(-), Leu(-)), obtaining three good Arg(+) transformants. These transformants [ChTF5 2(CYP1A1), ChTF5-4(CYP1A1), and ChTF5-6(CYP1A1)] were estimated to carry nine, six, and seven copies of the expression cassette on their chromosomes, respectively. Immunoblot analysis revealed that the three transformants produce similar amounts of rat CYP1A1 enzyme. ChTF5-2(CYP1A1), ChTF5-4(CYP1A1), ChTF5 6(CYP1A1) and recipient OJ1078 were cultivated in a liquid medium containing 2,7/2,8(at a ratio of 1:1)-dichlorodibenzo-p-dioxins (2,7/2,8-DCDDs) and the amount of intra- and extracellular 2,7/2,8-DCDDs remaining was measured. The results showed that all three transformants efficiently transform 2,7/2,8-DCDDs through the action of the recombinant rat CYP1A1 enzyme. PMID- 15812644 TI - [ABO blood group expression in corneal allograft failures]. AB - BACKGROUND: ABO blood group antigens are only expressed by the epithelial cells of normal human corneas. Since AB0 blood group antigens are also known to be expressed on stromal and endothelial cells of inflamed corneas, this study aimed to investigate the extend of ABO blood group antigen expression in corneal allograft failures. MATERIAL AND METHODS: Twenty-two failed corneal allografts of 22 patients were examined. In 12 cases the patients had clinically proven corneal allograft rejection. In 10 cases there was no evident history of allograft rejection and the diagnosis graft failure due to chronic endothelial cell loss was made. Immunohistochemical staining of paraffin embedded sections was performed with monoclonal mouse antibodies to human blood group antigen A or B using the streptavidin-biotin-peroxidase complex technique. RESULTS: Blood group antigens A or B were expressed by stromal keratocytes in 5 out of 12 and by endothelial cells in 7 out of 12 corneas with clinically proven immunologic graft rejection. Corneal transplants with chronic endothelial cell loss expressed blood group antigens A and/or B on the endothelial cells in three out of ten cases. CONCLUSION: The results of this study show that ABO blood group antigens can be up-regulated in cases of corneal allograft failure, especially in cases of immune mediated graft rejection. This phenomenon might play a role in corneal allograft rejection. PMID- 15812645 TI - Characterization of gene expression profiles of T cells during anti-tumor response. AB - BACKGROUND AND AIMS: T cells of tumor-bearing mice or cancer patients exhibit an immune dysfunction, enabling the tumor to escape immune surveillance. METHODS: The experiments are based on EL4 thymoma cells that were transfected with costimulatory ligands B7-1, B7-2, or both at the same time. We used oligonucleotide-based DNA chip microarrays to characterize the genomic expression profile of peripheral T cells according to their anti-tumor immune response in vivo. These murine T cells were also characterized by ELISA, FACS analysis, and co-stimulatory assays. RESULTS: Using commonly established methods, such as FACS analysis or the analysis of the cytokine profile by ELISA, it was not possible to determine functional differences in the in vivo activity of T lymphocytes against tumor cells. EL4 tumor cells induced multiple anti-tumor immune responses in vivo depending on their B7 expression. We successfully used microarray analysis to identify genes that were differentially expressed in the dysfunctional T cells, which were unable to reject tumors in vivo. Although Th1 and Th2 cytokine expression was not affected, we observed differential expression of genes involved in the regulation of an innate immune response. CONCLUSION: Our results provide evidence that the anti-tumor response can be identified by the "gene profile" of T cells. Genomic scale analysis offers the opportunity to identify subtle changes in gene expression in T cells reflecting a distinct biological behavior in vivo. PMID- 15812646 TI - The effect of hypophysectomy on pancreatic islet hormone and insulin-like growth factor I content and mRNA expression in rat. AB - The growth arrest after hypophysectomy in rats is mainly due to growth hormone (GH) deficiency because replacement of GH or insulin-like growth factor (IGF) I, the mediator of GH action, leads to resumption of growth despite the lack of other pituitary hormones. Hypophysectomized (hypox) rats have, therefore, often been used to study metabolic consequences of GH deficiency and its effects on tissues concerned with growth. The present study was undertaken to assess the effects of hypophysectomy on the serum and pancreatic levels of the three major islet hormones insulin, glucagon, and somatostatin, as well as on IGF-I. Immunohistochemistry (IHC), in situ hybridization (ISH), radioimmunoassays (RIA), and Northern blot analysis were used to localize and quantify the hormones in the pancreas at the peptide and mRNA levels. IHC showed slightly decreased insulin levels in the beta cells of hypox compared with normal, age-matched rats whereas glucagon in alpha cells and somatostatin in delta cells showed increase. IGF-I, which localized to alpha cells, showed decrease. ISH detected a slightly higher expression of insulin mRNA and markedly stronger signals for glucagon and somatostatin mRNA in the islets of hypox rats. Serum glucose concentrations did not differ between the two groups although serum insulin and C-peptide were lower and serum glucagon was higher in the hypox animals. These changes were accompanied by a more than tenfold drop in serum IGF-I. The pancreatic insulin content per gram of tissue was not significantly different in hypox and normal rats. Pancreatic glucagon and somatostatin per gram of tissue were higher in the hypox animals. The pancreatic IGF-I content of hypox rats was significantly reduced. Northern blot analysis gave a 2.6-, 4.5-, and 2.2-fold increase in pancreatic insulin, glucagon, and somatostatin mRNA levels, respectively, in hypox rats, and a 2.3-fold decrease in IGF-I mRNA levels. Our results show that the fall of serum IGF-I after hypophysectomy is accompanied by a decrease in pancreatic IGF-I peptide and mRNA but by partly discordant changes in the serum concentrations of insulin and glucagon and the islet peptide and/or mRNA content of the three major islet hormones. It appears that GH deficiency resulting in a "low IGF-I state" affects translational efficiency of these hormones as well as their secretory responses. The maintenance of normoglycemia in the presence of reduced insulin and elevated glucagon serum levels, both of which would be expected to raise blood glucose, may result mainly from the enhanced insulin sensitivity, possibly due to GH deficiency and the subsequent decrease in IGF-I production. PMID- 15812647 TI - Human dendritic cells process and present Listeria antigens for in vitro priming of autologous CD4+ T lymphocytes. AB - The role of human dendritic cells (DC) in the immune response toward intracellularly growing Listeria was analyzed under in vitro conditions using several morphological and functional methods. DC incubated with Listeria innocua and L. monocytogenes, respectively, readily phagocytosed the bacteria. Listeria did not impair viability and immunogenic potential of human DC. Listerial antigens were found to be processed within the lysosomal compartment of DC and colocalized with major histocompatibility complex (MHC) class II molecules, as shown by fluorescence and transmission electron microscopy. DC challenged with apathogenic L. innocua were highly effective in priming autologous naive T cells (mainly CD4+) in vitro. The T cells strongly proliferated in the presence of DC incubated with L. innocua, which could be significantly inhibited by anti-MHC II mAb. L. innocua-primed T cells were also successfully stimulated by DC harboring the pathogenic L. monocytogenes, either the wild-type strain EGD or the p60 reduced mutant strain RIII. From our results, we conclude that human DC infected with nonpathogenic intracellular bacteria are able to efficiently prime naive T cells, which are then suitable for recognition of antigens derived from related virulent bacterial species. This in vitro human model provides an interesting tool for basic research in infectious immunology and possibly for a new immunotherapy. PMID- 15812649 TI - Characterization by suppression subtractive hybridization of transcripts that are differentially expressed in leaves of apple scab-resistant and susceptible cultivars of Malus domestica. AB - In order to compare transcription profiles in cultivars of Malus domestica that are differentially sensitive to apple scab (Venturia inaequalis), two cDNA libraries were constructed using the suppression subtractive hybridization (SSH) method. Subtraction hybridization was performed between cDNAs from uninfected young leaves of the resistant cultivar Remo and the susceptible Elstar. In total, 480 EST clones were obtained: 218 (ELSTAR) clones represent transcripts that are preferentially expressed in Elstar, while the other 262 (REMO) are derived from RNAs that are more highly expressed in Remo. The putative functions of about 50% of the cloned sequences could be identified by sequencing and subsequent homology searches in databases or by dot-blot hybridization to known targets. In the resistant cv. Remo the levels of transcripts encoding a number of proteins related to plant defense (such as beta-1,3-glucanase, ribonuclease-like PR10, cysteine protease inhibitor, endochitinase, ferrochelatase, and ADP-ribosylation factor) or detoxification of reactive oxygen species (such as superoxide dismutase) were highly up-regulated relative to the amounts present in cv. Elstar. Most surprising was the large number of clones derived from mRNAs for metallothioneins of type 3 (91 out of 262) found in the REMO population. The corresponding transcripts were only present in small amounts in young uninfected leaves of the cv. Elstar, but were up-regulated in the susceptible cultivar after inoculation with V. inaequalis. These results indicate that constitutively high level expression of PR proteins may protect cv. Remo from infection by different plant pathogens. PMID- 15812648 TI - Guanylin-immunoreactive cells in the female and male rat adenohypophysis and their changes under various physiological and experimental conditions. AB - The peptide guanylin, first isolated from rat small intestine, is involved in the regulation of water-electrolyte transport between the intracellular and extracellular compartments of the epithelia. The main sites of guanylin expression are the intestinal, airway, or exocrine gland ductal epithelia where guanylin acts in a paracrine/luminocrine fashion. Because guanylin also circulates in the blood, sources of this peptide were sought in endocrine glands. Our group has already demonstrated the presence of guanylin-immunoreactive cells in the pars tuberalis of male rat adenohypophysis. In this study, we investigated whether guanylin-immunoreactive cells exist also in the adenohypophysial pars distalis and whether their appearance or distribution correlates with various physiological conditions in female rats or alters after gonadectomy in both sexes. These studies revealed that the rat pars distalis contains two guanylin immunoreactive cell types, gonadotrophic cells, whose number varied notably during the estrous cycle, reached a peak in the proestrous phase, and increased consistently during pregnancy, in lactating animals, and after gonadectomy, and folliculo-stellate cells, a discrete number of which were found only in female rats at the estrous phase. These findings suggest that guanylin is involved in regulating gonadotrophic cell function. They also add important information on the controversially discussed functions of folliculo-stellate cells. PMID- 15812650 TI - Characterisation of the Aspergillus niger dapB gene, which encodes a novel fungal type IV dipeptidyl aminopeptidase. AB - We have cloned the Aspergillus niger dapB gene. Analysis of its nucleotide sequence and the corresponding protein sequence indicates that the gene encodes a type IV dipeptidyl aminopeptidase (DPP IV). Based upon its deduced sequence we predict the presence of a transmembrane domain in the protein. Furthermore, dapB overexpressing transformants display an increase in intracellular DPP IV activity. This is the first reported characterisation of a dipeptidyl aminopeptidase with a transmembrane domain from a filamentous fungus. Using the dapB sequence as a query, we were able to identify 14 DPP IV-encoding genes, and 12 additional DPPIV proteases in public genomic databases. Phylogenetic analysis reveals that in yeasts there are two clades of genes that encode DPP IV proteases with a transmembrane domain. In this study we demonstrate that, as in yeasts, two classes of DPP IV-encoding genes exist in filamentous fungi. However, only one of these codes for DPP IV proteases with a transmembrane domain. The second type present in filamentous fungi encodes extracellular DPP IV proteases. The dapB gene belongs to the first cluster. We propose that DapB plays a role in the proteolytic maturation of enzymes produced by A. niger. PMID- 15812652 TI - Quality of life and a symptom cluster associated with breast cancer treatment related lymphedema. AB - OBJECTIVES: The aim of this study was to compare quality of life and symptoms between breast cancer survivors who have developed and undergone treatment for chronic lymphedema with those who have not developed lymphedema. PATIENTS AND METHODS: The cross-sectional, mixed-methods design included 64 breast cancer survivors with lymphedema and 64 breast cancer survivors without lymphedema. Variables assessed quantitatively included sociodemographic information, medical data, body mass index (BMI), arm extracellular fluid volume, quality of life (QOL), and physical and emotional symptoms. For the qualitative component, individuals with lymphedema responded in writing to the question: During the past week what other difficulties have you experienced because of your lymphedema? RESULTS: Compared with those without lymphedema, breast cancer survivors with lymphedema reported poorer QOL. A symptom cluster that included alteration in limb sensation, loss of confidence in body, decreased physical activity, fatigue, and psychological distress was identified. Perception of limb size influenced the cumulative symptom experience more than objective arm volume. Qualitative data revealed multiple QOL, physical health, and psychological concerns. BMI correlated with multiple outcomes. CONCLUSIONS: Findings suggest that current lymphedema treatments, although beneficial, may not provide complete relief of symptoms associated with lymphedema and complementary interventions are needed. The poorer QOL in breast cancer survivors with lymphedema may relate to the presence of an untreated symptom cluster. PMID- 15812655 TI - Inter-annual variability of NDVI in response to long-term warming and fertilization in wet sedge and tussock tundra. AB - This study explores the relationship between the normalized difference vegetation index (NDVI) and aboveground plant biomass for tussock tundra vegetation and compares it to a previously established NDVI-biomass relationship for wet sedge tundra vegetation. In addition, we explore inter-annual variation in NDVI in both these contrasting vegetation communities. All measurements were taken across long term experimental treatments in wet sedge and tussock tundra communities at the Toolik Lake Long Term Ecological Research (LTER) site, in northern Alaska. Over 15 years (for wet sedge tundra) and 14 years (for tussock tundra), N and P were applied in factorial experiments (N, P and N+P), air temperature was increased using greenhouses with and without N+P fertilizer, and light intensity was reduced by 50% using shade cloth. during the peak growing seasons of 2001, 2002, and 2003, NDVI measurements were made in both the wet sedge and tussock tundra experimental treatment plots, creating a 3-year time series of inter-annual variation in NDVI. We found that: (1) across all tussock experimental tundra treatments, NDVI is correlated with aboveground plant biomass (r2 = 0.59); (2) NDVI-biomass relationships for tussock and wet sedge tundra communities are community specific, and; (3) NDVI values for tussock tundra communities are typically, but not always, greater than for wet sedge tundra communities across all experimental treatments. We suggest that differences between the response of wet sedge and tussock tundra communities in the same experimental treatments result from the contrasting degree of heterogeneity in species and functional types that characterize each of these Arctic tundra vegetation communities. PMID- 15812653 TI - Factors influencing preferences for place of terminal care and of death among cancer patients and their families in Korea. AB - GOALS OF WORK: This study examined cancer patient and family member preferences and the reasons for the preferences-regarding place of terminal care and of death. PATIENTS AND METHODS: We constructed a questionnaire that included demographic, clinical, and support network data for 371 patients who were treated at any of the seven university hospitals or the National Cancer Center in Korea and 281 of their family members. MAIN RESULTS: About half of the interviewed patients and half of the family members expressed a preference for the patient being cared for and dying at home. The preference reflected a wish for patients to live out their lives in privacy and to be with their family when their life ended. Those who expressed a preference to be cared for or to die in a hospital wanted to get medical treatment during the last days of life and to relieve their families of the burden of caring for them. Of the variables examined, support network factors and some sociodemographic factors (sex, family members' age, and place of residence) were strongly predictive of preferences. CONCLUSION: A majority of cancer patients preferred to receive terminal care at home. Cancer patients and family members with strong support groups were more likely to prefer the home as the place of terminal care and dying. Hence, improving support networks might increase the proportion of patients receiving home care and dying at home. PMID- 15812656 TI - Post-continuous whole body exposure of rabbits to 650 MHz electromagnetic fields: effects on liver, spleen, and brain. AB - This study deals with the effects of post (12 or 18 months) whole body continuous (2 years) exposure of rabbits to 650 MHz electromagnetic fields (EMF) that are characteristic of those produced by broadcasting stations, on body weight and body mass, on the morphology of liver, spleen and brain, and on apoptosis rates and glycogen distribution in the liver. Two groups of rabbits were continuously exposed for 2 years to EMF of 650 MHz followed by 12 months (group 1) or 18 months (group 2) of post-exposure; a third group (group 3) was sham exposed. It was shown that the changes in apoptosis rates were conditional during the time of exposure, but not on a specific organ and that the whole body continuous exposure gave rise to modifications whose types and intensities were related to the time of post-exposure (12 or 18 months, respectively), the type of organ, and the individual animal. A number of effects were observed only in group 1, and not in group 2, which suggests some kind of adaptive response or of long-term recovery in the rabbits following continuous exposure to 650 MHz EMF. PMID- 15812657 TI - Is there reliable experimental evidence for different dicentric yields in human lymphocytes produced by mammography X-rays free-in-air and within a phantom? AB - We examined the production of dicentrics in human lymphocytes irradiated with 29 kV X-rays to a depth of 13.5 mm in a PMMA phantom. For these irradiation conditions, which are appropriate for the diagnostic application of mammography X rays, a coefficient alpha of (5.88+/-0.66) x 10(-2) Gy(-1) of the linear quadratic dose-response relationship was determined. This value does not differ significantly from the coefficient alpha of (6.55+/-0.97) x 10(-2) Gy(-1) obtained earlier for a free-in-air set-up using blood of the same donor. The results are interpreted in terms of both the energy distributions of the photon fluence of mammography X-rays free-in-air and those in the PMMA phantom. Based on earlier results of experiments with monochromatic X-rays in the energy range 1.83 40 keV (completed here by an additional measurement at 25 keV), a fit function alpha(E) to the measured alpha coefficients as a function of the energy E of monochromatic X-rays was used to calculate weighted mean values alpha for both the mammography X-ray spectra free-in-air and in the phantom. As a result, weighted mean values of (4.9+/-1.0) x 10(-2) Gy(-1) and (4.5+/-1.0) x 10(-2) Gy( 1) were obtained, respectively. Although the measured alpha coefficients for mammography X-rays appear to be systematically higher than those calculated as weighted mean values alpha, it can be concluded that the modification of the mammography X-ray spectrum to a depth of 13.5 mm in a PMMA phantom compared with the free-in-air spectrum has no significant influence on the dicentric yields in human lymphocytes. PMID- 15812658 TI - Transgenic plants of Vitis vinifera cv. Seyval blanc. AB - Leaf discs of grapevine cv. Seyval blanc originating from in vitro cultures were transformed with Agrobacterium tumefaciens strain LBA 4404 harbouring the vector pGJ42 carrying genes for chitinase and RIP (ribosome-inactivating protein) in an attempt to improve fungal resistance. The gene for neomycin phosphotransferase II (nptII) was used as the selectable marker gene. The explants were cocultivated for 2 days with recombinant Agrobacteria and then submitted to selection on NN69 medium containing 100 mg/l kanamycin. Successful regeneration and conversion of transgenic plantlets were obtained. Stable integration of foreign DNA was confirmed by PCR and Southern blot analyses, and protein expression was detected by Western blot. The regenerated transgenic plants were adapted to the greenhouse and showed no evidence of phenotypical alterations. The foreign genes introduced into the transformed plants did not effect the expected improvement in fungal disease resistance under field conditions for the major pests Uncinula necator and Plasmopara viticola. PMID- 15812659 TI - Effective selection of transgenic papaya plants with the PMI/Man selection system. AB - The selectable marker gene phospho-mannose isomerase (pmi), which encodes the enzyme phospho-mannose isomerase (PMI) to enable selection of transformed cell lines on media containing mannose (Man), was evaluated for genetic transformation of papaya (Carica papaya L.). We found that papaya embryogenic calli have little or no PMI activity and cannot utilize Man as a carbon source; however, when calli were transformed with a pmi gene, the PMI activity was greatly increased and they could utilize Man as efficiently as sucrose. Plants regenerated from selected callus lines also exhibited PMI activity but at a lower specific activity level. Our transformation efficiency with Man selection was higher than that reported using antibiotic selection or with a visual marker. For papaya, the PMI/Man selection system for producing transgenic plants is a highly efficient addition to previously published methods for selection and may facilitate the stacking of multiple transgenes of interest. Additionally, since the PMI/Man selection system does not involve antibiotic or herbicide resistance genes, its use might reduce environmental concerns about the potential flow of those genes into related plant populations. PMID- 15812660 TI - The promoter of Milk vetch dwarf virus component 8 confers effective gene expression in both dicot and monocot plants. AB - The activity of a predicted promoter, PMC8, from Milk vetch dwarf virus was evaluated by comparing it with the cauliflower mosaic virus 35S RNA promoter (P35S) and PNCR, a promoter from Soybean chlorotic mottle virus. When the GUS fusion gene was introduced into tobacco, PMC8 showed a similar expression profile to P35S but with a more intense expression in proliferating tissues. The usefulness of PMC8 was confirmed by driving NPTII for selection of kanamycin resistant tobacco plants with improved transformation efficiency. PMC8 was also effective in transgenic rice plants. Thus, PMC8 is useful as an alternative to P35S in both dicotyledonous and monocotyledonous plants, especially for gene expression in proliferating tissues. PMID- 15812661 TI - Mitotic instability in wheat x Thinopyrum ponticum derivatives revealed by chromosome counting, nuclear DNA content and histone H3 phosphorylation pattern. AB - To evaluate the mitotic stability of Triticum aestivum x Thinopyrum ponticum derivatives (BC(2)F(7) and BC(2)F(5) doubled haploids), chromosome counting by both conventional and immunostaining techniques, and measurement of DNA content were performed. The wheat progenitor line, PF 839197, the wheat recurrent parent CEP 19 and the control Chinese Spring were also investigated. In the hybrid derivatives, chromosome number ranged from 2n=36 to 60, with a predominance of chromosome numbers higher than 2n=42, that was confirmed by determination of nuclear DNA content. Chinese Spring' and PF 839197 were stable, but CEP 19 showed chromosome number variation (20%). Analyses of non-pretreated cells revealed the presence of anaphase bridges, lagging chromatids, chromosome fragments and micronuclei. Immunostaining with an antibody recognizing histone H3 phosphorylated showed dicentric chromatids forming anaphase bridges and pericentromeric phosphorylation at centric chromosome fragments but not at lagging chromatids. The possible causes of the observed mitotic instability are discussed. PMID- 15812662 TI - Cutis marmorata telangiectatica congenita with gangrenous ulceration and hypovolaemic shock. AB - Cutis marmorata telangiectatica congenita (CMTC) is an uncommon sporadic congenital vascular anomaly characterised by persistent cutis marmorata, telangiectasia, and phlebectasia. The disease usually has a benign clinical course and over 50% of cases recover spontaneously without specific treatment. The occurrence of life-threatening complications in CMTC is rare. CONCLUSION: We report a neonate with life-threatening cutis marmorata telangiectatica congenita which was complicated by gangrenous ulceration, bleeding episodes, and hypovolaemic shock. PMID- 15812663 TI - A retrospective study of the demographics of sport and exercise injuries in 1143 children presenting to an Irish emergency department over a 6-month period. AB - The purpose of this study was to provide up-to-date data on the nature of sport related injury (SRI) presenting to a large emergency department in Ireland. Data were collected retrospectively on all children under 17 years of age with a SRI, presenting to the emergency department of a major teaching hospital, over a 6 month period, and entered into a Microsoft Access database. A total of 1143 SRIs were identified which had occurred over a 6-month period, from 53 different sports. There was a high proportion of humerus and back SRIs in females, and a higher proportion of falls in females. Males were more frequently involved in collisions. Children with SRI were not using protective equipment in 94% of cases. Advice regarding rest, ice, compression and elevation (RICE)/general injury advice was given to 25% of patients and regarding injury preventive measures in less than 0.1% of cases. Of children, 28% had previously attended with a SRI. We also observed a lower rate of analgesia prescription to children under age 4, compared to children of an older age, and rarity of topical analgesic prescription. Overall, 10% of SRIs required admission, with 65% of these cases needing orthopaedic intervention. CONCLUSION: The data provided from this study should raise awareness of the different aspects of sport related injuries affecting children, and may help to provide the impetus for suggesting direction and guidance for reducing such events. PMID- 15812664 TI - Acute hypereosinophilia associated with right ventricular thrombosis. PMID- 15812665 TI - Somatosensory evoked potentials in children with severe head trauma. AB - We evaluated the predictive value of somatosensory evoked potentials (SEP) in a series of children with severe traumatic brain injury (TBI). The prospective clinical investigation was performed in a Level I paediatric trauma centre. We included 26 consecutive comatose paediatric patients aged from 1 month to 17 years (median age 11 years) following severe TBI (initial Glasgow Coma Score (GCS) 8 or below). Besides SEP recordings, the intracranial pressure and the results of an initial cranial CT scan were filed. The Glasgow Outcome Scale (GOS) was used to assess outcome at discharge. Thirteen children had normal SEP measurements, three patients had abnormal SEP recordings and a cortical response was bilaterally absent in ten children. Out of 26 children, 10 died whereas two remained in a persistent vegetative state. Only one child suffered from significant neurological deficits (GOS 3) at discharge. Seven patients survived with a GOS of 4 and six children survived without neurological impairment (GOS 5). Normal SEP indicated a favourable outcome in most children but did not rule out the occurrence of death, while absence of SEP was related to unfavourable outcome in all cases. CONCLUSION: Measurement of somatosensory evoked potentials provides valuable data for determining the prognosis at early coma stages. Our data show that an unfavourable outcome can be predicted with higher precision than a favourable outcome. PMID- 15812666 TI - Interferon-gamma production by human cord blood monocyte-derived dendritic cells. AB - Interferon (IFN)-gamma is produced by T cells and natural killer cells and activates monocytes and dendritic cells (DCs). Recently, IFN-gamma has been shown to be produced by mouse DCs following stimulation with interleukin (IL)-12, which is markedly augmented by the addition of IL-18. We here analyzed whether human DCs secrete IFN-gamma in response to IL-12 and/or IL-18. Human immature DCs, generated from cord blood CD14(+) monocytes by treating with granulocyte macrophage colony-stimulating factor and IL-4, were incubated with IL-12 and/or IL-18 and assayed for IFN-gamma production. IL-12, but not IL-18, weakly induced IFN-gamma production, while IL-12 together with IL-18 induced high levels of IFN gamma production. Similar results were obtained with mature DCs, although levels of IFN-gamma production were less than those in immature DCs. Also with mature and immature DCs, IL-12 upregulated the expression of IL-18 receptor alpha (Ralpha), and costimulation with IL-12 and IL-18 upregulated CD40 expression. Anti-IL-18Ralpha antibody abrogated both the IFN-gamma induction and the CD40 upregulation by IL-12 plus IL-18. These findings suggest that IL-12 upregulates IL-18Ralpha expression on human DCs and acts synergistically with IL-18 to induce high levels of IFN-gamma, which subsequently enhances CD40 expression on DCs in an autocrine manner. PMID- 15812667 TI - Phenology of temperate trees in tropical climates. AB - Several North American broad-leaved tree species range from the northern United States at approximately 47 degrees N to moist tropical montane forests in Mexico and Central America at 15-20 degrees N. Along this gradient the average minimum temperatures of the coldest month (T (Jan)), which characterize annual variation in temperature, increase from -10 to 12 degrees C and tree phenology changes from deciduous to leaf-exchanging or evergreen in the southern range with a year-long growing season. Between 30 and 45 degrees N, the time of bud break is highly correlated with T (Jan) and bud break can be reliably predicted for the week in which mean minimum temperature rises to 7 degrees C. Temperature-dependent deciduous phenology-and hence the validity of temperature-driven phenology models terminates in southern North America near 30 degrees N, where T (Jan)>7 degrees C enables growth of tropical trees and cultivation of frost-sensitive citrus fruits. In tropical climates most temperate broad-leaved species exchange old for new leaves within a few weeks in January-February, i.e., their phenology becomes similar to that of tropical leaf-exchanging species. Leaf buds of the southern ecotypes of these temperate species are therefore not winter-dormant and have no chilling requirement. As in many tropical trees, bud break of Celtis, Quercus and Fagus growing in warm climates is induced in early spring by increasing daylength. In tropical climates vegetative phenology is determined mainly by leaf longevity, seasonal variation in water stress and day length. As water stress during the dry season varies widely with soil water storage, climate-driven models cannot predict tree phenology in the tropics and tropical tree phenology does not constitute a useful indicator of global warming. PMID- 15812668 TI - [Age and survival likelihood of polytrauma patients. "Local tailoring" of the DGU prognosis model]. AB - INTRODUCTION: Age is one of five prognostic parameters identified based on data of the trauma registry of the German Association for Trauma Surgery (DGU). We asked ourselves if the suggested prognostic model provides the same predictive power of data from an independent hospital. Furthermore, we investigated whether age itself or age-associated comorbidity causes an unfavorable prognostic effect. METHODS: The investigation was based on data of 103 multiply injured patients (67 male, 36 female, mean age 35,4+/-SD 19,0 years, ISS 36,8+/-10,9). Data were collected prospectively following the guidelines of the trauma registry of the German Association for Trauma Surgery. Based on documented comorbidities, a risk calculation was performed using the ASA classification. Correlation between age and ASA was analyzed using Spearman's method. The prognostic value of the original model in our patient pool with or without ASA classification, possible interactions, and the discriminatory power of the model were estimated using logistic regression. RESULTS: Attributable mortality was 31,7% (95% CI 22,7 41,7%). Age, ISS, GCS and ASA were included into the final logistic model. Odds ratios of the origin model were reproducible nearly identical in our patinet pool (OR: age 1,048; ISS 1,066; GCS 0,822). In spite of the fact that we have found a strong correlation between age and ASA-Classification (rho=0,60, p<0,0001) there was no prognostic value of comorbidity. CONCLUSION: The suggested prognostic model based on multicenter data evaluation can be applied to a single center with only minimal loss of discriminatory power. In this context, age seems to have a prognostic value independent of comorbidity. PMID- 15812669 TI - Uptake of [3H]-gangliosides by an intestinal protozoan, Giardia lamblia. AB - Because of their limited lipid synthesis ability it has been postulated that Giardia lamblia trophozoites depend on lipid remodeling reactions, to generate parasite-specific phospho and glycolipids. We have shown earlier that exogenous bile acids and lipid molecules are taken up by Giardia through active transport and by other mechanisms. Another report suggests that lipoprotein-like receptors may be present in this parasite that are involved in lipid endocytosis. In the present investigation, we asked whether and by what mechanism exogenous gangliosides can be taken up by Giardia. We found that ganglioside uptake was dependent, on time and concentration, but was not temperature- or carrier dependent. Gangliosides encapsulated into liposomes were taken up much more effectively than non-encapsulated gangliosides. These results are consistent with a passive uptake process, facilitated by membrane fusion with lipid carriers, but not dependent on a catalytic carrier mechanism. This observation may have some therapeutic implications. PMID- 15812670 TI - Phylogenetic position of Dracunculus medinensis and some related nematodes inferred from 18S rRNA. PMID- 15812671 TI - Development of Taenia saginata asiatica metacestodes in SCID mice and its infectivity in human and alternative definitive hosts. AB - Development of Taenia saginata asiatica metacestodes in SCID mice, and its infectivity in humans, golden hamsters, and Mongolian gerbils as alternative definitive hosts, were investigated. Cysticerci were recovered from SCID mice that were subcutaneously injected with hatched oncospheres of T. s. asiatica. The morphological changes of metacestodes were observed. The recovered cysticerci were fed to gerbils, hamsters and humans, to check for their infectivity. Tapeworms were recovered from gerbils and hamsters fed with 20 to 45 week-old cysticerci, and proglottids excretions were observed in human volunteers fed with 45 week-old cysticerci. However, no tapeworms were recovered from gerbils fed with 10 week-old cysticerci. Our results suggest that T. s. asiatica oncospheres needed more than 20 weeks to develop to maturity in SCID mice to be infective to both their natural and alternative definitive hosts. PMID- 15812672 TI - Plasmodium (Haemamoeba) cathemerium gene sequences for phylogenetic analysis of malaria parasites. AB - The DNA sequence information on avian malaria parasites of the genus Plasmodium is quite limited. At present, sequences of only 6 out of 34 valid species are available. However, sequence data of avian malaria parasites are particularly important with regard to the resolution of the phylogenetic relationships of the most virulent human malaria agent, Plasmodium falciparum. The question as to whether P. falciparum originates from avian or from mammalian parasites would contribute to our understanding of its biology and would probably facilitate the interpretation of experimental results. To add to the body of molecular data, we sequenced three genes (cytochrome b, 18 SSU rRNA, caseinolytic protease C) of different organellar origin of one of the most widespread avian malaria parasites, Plasmodium (Haemamoeba) cathemerium, which once used to be an important laboratory in vivo model in human malaria research. The analysis of the new P. cathemerium sequences in direct comparison with the rodent parasite P. berghei and the four human malaria parasites by pairwise distance calculation do not suggest a closer relationship of P. cathemerium to P. falciparum than to the other species involved. PMID- 15812673 TI - Helminth parasites in freshwater fish from the Papaloapan river basin, Mexico. AB - A checklist based on previously published records and original data is presented for the helminth parasites reported in 35 fish species from nine families from the Rio Papaloapan basin, east Mexico. The checklist contains 85 taxa from 39 helminth families. Trematodes and nematodes were the most abundant taxonomic groups. The helminth fauna in the fish of the Papaloapan River basin predominantly consists of Neotropical species that are largely autogenic. The introduced species Centrocestus formosanus was the most widely distributed helminth, infecting 16 host species. Ten of the recorded helminth species have only been found in fish from the Papaloapan. This inventory contributes 157 new host records, and reports the presence of 30 helminth species in the Papaloapan for the first time . This inventory shows the richness of helminth parasite species in the fish of the Papaloapan River basin in comparison with the other hydrological basins in Mexico. It also demonstrates that this fauna is typically Neotropical and quite similar to that from the neighboring basins of the Grijalva Usumacinta system and the Yucatan Peninsula. The data also suggest highly effective transmission between environments within the same basin and that the regional parasite fauna is strongly influenced by fish community composition. PMID- 15812674 TI - HERG K+ channel expression-related chemosensitivity in cancer cells and its modulation by erythromycin. AB - PURPOSE: Previous studies have found that the HERG K+ channel is highly expressed in some cancers. In the study reported here, we investigated HERG expression in various cancer cell lines, its correlation with chemosensitivity to vincristine, paclitaxel, and hydroxy-camptothecin, and its biochemical modulation. METHODS: The MTT assay and clonogenic assay were used to detect the cytotoxicity of anticancer drugs in vitro. HERG expression was analyzed by Western blotting or immunocytochemistry. Gene transfection was used to examine the changes in HERG related chemosensitivity. Cell cycle phase distribution was detected by flow cytometry and drug combinations were evaluated by the MTT assay. RESULTS: HERG expression levels differed widely between various human cancer cell lines and HT 29 cells expressing high levels of HERG were more sensitive than A549 cells expressing low levels of HERG to vincristine, paclitaxel, and hydroxy camptothecin. In terms of IC50, the chemosensitivities of herg-transfected A549 cells to vincristine, paclitaxel and hydroxy-camptothecin were significantly increased. However, for cisplatin and 5-fluorouracil, no significant difference between herg-transfected A549 cells and parent A549 cells was detected. Erythromycin, a HERG K+ channel blocker, suppressed the growth of various cancer cells and the potency was correlated with HERG expression levels. Combinations of erythromycin and vincristine, paclitaxel or hydroxy-camptothecin showed synergy in cytotoxicity to HT-29 cells. Erythromycin also enhanced the G2/M arrest induced by vincristine in HT-29 cells. There were synergistic effects between erythromycin and vincristine, paclitaxel, and hydroxy-camptothecin, and chemosensitivity was correlated with HERG expression level. CONCLUSIONS: HERG expression levels and chemosensitivity were positively correlated for vincristine, paclitaxel, and hydroxy-camptothecin. Erythromycin was active as a modulator. These results suggest that HERG may serve as a molecular marker and modulating target for individualized cancer therapy. PMID- 15812675 TI - [Blocking adhesion molecules with natalizumab in multiple sclerosis]. AB - Natalizumab is a humanized, monoclonal antibody, that inhibits adhesion molecules (alpha(4)-integrins) on the surface of immune cells. These adhesion molecules are important for binding of lymphocytes to endothelial cells of blood vessels and infiltration of inflammatory cells into tissues. Natalizumab is currently being tested in large clinical trials for the treatment of multiple sclerosis (MS) and other autoimmune diseases (inflammatory bowel diseases, rheumatoid arthritis). After demonstrating the safety and potential effectiveness of natalizumab in MS therapy during shorter treatment periods (50%, were enrolled in a 16-week, open-label trial of alagebrium 420 mg per day. Assessments included: peak exercise oxygen consumption, aortic distensibility, and left ventricular EF and mass by magnetic resonance imaging, Doppler diastolic filling, and quality of life by the Minnesota Living with Heart Failure questionnaire. One patient discontinued treatment because of a myocardial infarction after 12 days of treatment, and a second died suddenly after 10 weeks of treatment. Thus 21 patients completed the study. Left ventricular mass was 124 +/- 35 g at baseline and decreased to 119 +/ 34 g at follow up ( P = .036). This was accompanied by a decrease in the ratio of Doppler early diastolic flow velocity to Doppler early diastolic mitral annulus velocity (E') from 10.6 +/- 2.7 to 9.4 +/- 1.9 ( P = .076) and an increase in E' from 7.3 +/- 1.2 to 8.4 +/- 1.7 cm/s ( P = .045). The Minnesota Living with Heart Failure total score improved from 41 +/- 21 to 32 +/- 21 ( P = .01). There were no changes in EF (64 +/- 4% at baseline), blood pressure, peak exercise oxygen consumption, and aortic distensibility. CONCLUSION: Sixteen weeks of treatment with the glucose crosslink breaker, alagebrium, resulted in a decrease in left ventricular mass and improvements in left ventricular diastolic filling and quality of life in patient with DHF. PMID- 15812747 TI - Treating diastolic heart failure with AGE crosslink breakers: thinking outside the heart failure box. PMID- 15812748 TI - Demographics, clinical characteristics, and outcomes of patients hospitalized for decompensated heart failure: observations from the IMPACT-HF registry. AB - BACKGROUND: Hospitalizations for decompensated heart failure are frequent. The Initiation Management Pre-discharge Assessment of Carvedilol Heart Failure (IMPACT-HF) registry collected observational data in patients hospitalized for worsening heart failure to characterize an unselected group of patients and to confirm the generalizability of the IMPACT-HF main trial population. METHODS AND RESULTS: The IMPACT-HF registry was conducted concurrently with the IMPACT-HF study, a randomized trial of in-hospital initiation of carvedilol compared with the standard practice of postdischarge beta-blocker initiation. Patients were eligible for registry enrollment if they were hospitalized for heart failure regardless of ejection fraction. There were no exclusions to participation. Patients were followed for 60 days. The IMPACT-HF Registry enrolled 567 patients. The mean age was 71 years, 52% of the patients were men and 82% were Caucasian. At discharge, 71% received an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker, 41% received digoxin, and 62% received beta blockers. The 60-day rate of rehospitalization or death was 31%. CONCLUSION: The IMPACT-HF registry enrolled elderly patients admitted for worsening heart failure primarily resulting from progressive volume overload. The 60-day rate of death or rehospitalization was high despite the use of evidence-based therapies. New treatments for this population are needed to decrease the morbidity and mortality associated with decompensated heart failure. PMID- 15812749 TI - Long-term results of coronary artery bypass grafting in patients with ischemic cardiomyopathy: the impact of renal insufficiency and noncardiac vascular disease. AB - BACKGROUND: We examined outcomes of patients with ischemic cardiomyopathy (ICMP), defined by left ventricular ejection fraction (LVEF) <35%, compared with patients with better-preserved LVEF, undergoing coronary bypass graft surgery (CABG). In addition, we examined the relative impact of a reduced LVEF in comparison with other comorbidities on long-term mortality in these patients. METHODS AND RESULTS: We evaluated 1381 patients (114 with ICMP, 1267 with better-preserved LVEF) who underwent isolated CABG at a tertiary Veterans Administration (VA) hospital between 1990 and 2000 using data from the VA Continuous Improvement in Cardiac Surgery Program and other VA databases. The 5-year survival was 74.0% in patients with ICMP and 84.4% in the group with better-preserved LVEF ( p = .005). LVEF <35% remained a significant predictor of long-term mortality in multivariable models (hazard ratio [HR] 1.55, 95% confidence interval [CI] 1.02 2.35). However, the presence of comorbidities, especially renal insufficiency, peripheral vascular disease, and cerebrovascular disease, had a similar or greater impact on long-term mortality. Renal insufficiency (serum creatinine >1.5 mg/dL) was associated with the highest risk of long-term mortality (HR 2.02, 95% CI 1.46-2.80). The use of a left internal thoracic artery graft reduced the risk of long-term mortality (HR 0.72, 95% CI 0.54-0.98). CONCLUSION: Even though severely depressed LVEF is associated with an increased risk of long-term mortality, the presence of comorbid factors, especially renal dysfunction and noncardiac vascular disease, increase the risk of long-term mortality by a similar or even larger magnitude. These comorbid factors should be given important consideration when evaluating the risks and benefits of CABG. PMID- 15812751 TI - Superoxide dismutase activity in adriamycin-induced cardiotoxicity in humans: a potential novel tool for risk stratification. AB - BACKGROUND: Oxidative stress has been implicated in Adriamycin cardiotoxicity experimentally. We evaluated whether changes in systemic markers of antioxidant reserve occur and are associated with left ventricular (LV) dysfunction during Adriamycin use in humans. METHODS AND RESULTS: We prospectively evaluated oncology patients eligible for Adriamycin chemotherapy. Blood samples for enzymatic (erythrocyte superoxide dismutase activity [SOD-U SOD/mg protein]) and nonenzymatic antioxidants (total radical trapping antioxidant potential [TRAP-U of Trolox/microL plasma]) were collected at baseline (B), intermediate (I), and final (F) cycles. LV ejection fraction (LVEF) was assessed by radionuclide ventriculography. Fifty-one patients (49 +/- 12 years, 90% female) underwent 5.9 +/- 0.9 chemotherapy cycles and received 301 +/- 52 mg/m 2 of Adriamycin. LVEF decreased from 61 +/- 6% (B) to 56 +/- 7% (F) ( P < .001), but only 6 (12%) patients developed significant LV systolic dysfunction (LVEF < 50%). SOD activity increased significantly during treatment (4.5 +/- 1.8 [B], 6.0 +/- 2.1 [I], 5.6 +/- 2.2 [F]; P < .01), whereas TRAP values were unchanged. Baseline SOD activity from patients who developed LV systolic dysfunction was significantly higher than from those who maintained normal LVEF (5.9 +/- 1.8 versus 4.3 +/- 1.7; P < .05). In multivariate analysis, baseline SOD levels remained independently associated with LV dysfunction ( P = .05). CONCLUSION: Erythrocyte SOD activity increases after Adriamycin treatment and high baseline levels predicts Adriamycin-induced cardiotoxicity in humans. PMID- 15812750 TI - Effects of different degrees of sympathetic antagonism on cytokine network in patients with ischemic dilated cardiomyopathy. AB - BACKGROUND: The proinflammatory cytokines have been implicated in the pathogenesis of heart failure. Recent studies have shown that beta-adrenergic blockade can modulate cytokine production. This study investigates the different impact of different degrees of sympathetic antagonism on circulating levels of cytokines in patients with heart failure resulting from ischemic dilated cardiomyopathy (IDC). METHODS AND RESULTS: Thirty-five patients with IDC were randomly assigned to receive metoprolol or carvedilol in an open-label study. Echocardiographic measurements and circulating levels of tumor necrosis (TNF) alpha and interleukin (IL)-1beta and IL-6 were obtained at baseline and after 3 months of treatment. The 2 beta-blockers significantly improved the left ventricular ejection fraction and reduced end-diastolic and end-systolic volume. The magnitude of these changes was greater with carvedilol than with metoprolol (respectively P < .001, P < .05, and P < .05). Both treatments induced a significant decrease in the levels of cytokines (for all P < .01), but the decrease in TNF-alpha and IL-1beta was more consistent in the carvedilol group ( P < .01). CONCLUSION: Our results support the hypothesis that a more complete block of sympathetic activity by carvedilol induces a greater decrease in the circulating levels of proinflammatory cytokines that could explain, at least in part, the better improvement in the left ventricular remodelling and systolic function in patients with IDC. PMID- 15812752 TI - Prolonged repolarization after ventricular assist device support is associated with arrhythmias in humans with congestive heart failure. AB - BACKGROUND: Recent observations indicate that the QTc interval often increases in the early postoperative period (<1 week) after mechanical unloading of severely failing hearts with a left ventricular assist device (LVAD). The present study examined whether early changes in ventricular repolarization after LVAD placement are associated with ventricular arrhythmias. METHODS AND RESULTS: An electrocardiogram was obtained within 4 days before LVAD placement, <12 hours after LVAD placement, and weekly thereafter. Patient records were reviewed for documented ventricular tachycardia (VT) or ventricular fibrillation (VF) for 1 week preoperatively and the first 2 weeks postoperatively. Differences in QTc interval between patients with and without VT were evaluated. Ten of 17 patients enrolled (59%) had VT or VF after LVAD placement. Of these, 4 required therapeutic intervention because of clinical instability or symptoms. The change in the QTc (DeltaQTc) between the preoperative and immediate postoperative period was significantly different among patients with VT/VF compared with patients without VT/VF (+23 ms vs. -68 ms, P < .001). CONCLUSION: The early period after initiation of LVAD support of the failing human heart is associated with a relatively high incidence of significant ventricular arrhythmias after LVAD placement. Beyond the impact of myocardial inflammation and wound healing occurring after all LVAD implants, early postoperative increases in the QTc interval after cardiac unloading appear to predispose to ventricular arrhythmias. PMID- 15812753 TI - A comparison of cardiac resynchronization by sequential biventricular pacing and left ventricular pacing to simultaneous biventricular pacing: rationale and design of the DECREASE-HF clinical trial. AB - BACKGROUND: The first generation of cardiac resynchronization therapy (CRT) devices approved for the treatment of heart failure used simultaneous biventricular (BiV) pacing to achieve ventricular resynchronization. Left ventricular pacing alone and sequential BiV pacing also show promise as alternative ways to deliver CRT, but have not been studied together in a large randomized trial. METHODS: The Device Evaluation of CONTAK RENEWAL 2 and EASYTRAK 2: Assessment of Safety and Effectiveness in Heart Failure (DECREASE-HF) Trial is a randomized, double-blind, 3-arm study of patients in New York Heart Association Class III or IV with an ejection fraction of 35% or less and a QRS duration > or =150 ms. Patients are randomized to receive either left ventricular pacing, simultaneous BiV pacing, or sequential BiV pacing. CONCLUSION: The study uses a novel composite endpoint that combines peak oxygen consumption and left ventricular end systolic dimension, thus combining a measure of symptomatic improvement (peak oxygen consumption) with a physiologic measure of ventricular reverse remodeling (left ventricular end systolic dimension) into a single composite score. Additionally, the safety and effectiveness of the CONTAK RENEWAL 2/4/4HE/EASYTRAK 2 system will be evaluated using: heart failure-related adverse events; system-related complications; left ventricular lead-related complications; detection time of induced ventricular fibrillation; and left ventricular lead performance (pacing threshold, pacing impedance, and R-wave amplitude). PMID- 15812754 TI - Treating heart failure with enhanced external counterpulsation (EECP): design of the Prospective Evaluation of EECP in Heart Failure (PEECH) trial. AB - BACKGROUND: Enhanced external counterpulsation (EECP) treatment can improve exercise tolerance in patients with ischemic heart disease; however, the possible benefits of EECP in patients with stable heart failure (HF) and left ventricular dysfunction (LVD) are unclear. An open pilot study showed significant increases in exercise tolerance in HF patients undergoing EECP. Thus a larger, controlled study of EECP in patients with stable HF (New York Heart Association [NYHA] classes II and III) and LVD was undertaken. METHODS AND RESULTS: The PEECH trial is a controlled, randomized, single-blind, parallel-group, multicenter study of 187 patients with symptomatic but stable HF (NYHA classes II and III) and an LV ejection fraction < or =35% was designed to assess the efficiency of EECP in patients with stable HF. Medical therapy is optimized in all patients based on the recommendations of the Heart Failure Society of America ("Usual Care"), and then randomized between 2 treatment groups; UC or EECP (35 hours over 7 weeks). CONCLUSION: Efficacy measures include standard exercise tolerance tests on a treadmill (modified Naughton protocol), with measurements of peak oxygen uptake and exercise duration time; quality of life questionnaires; NYHA classification; and neurohormonal markers of HF. PMID- 15812758 TI - Relationship of symptomatology with closed chamber infrared thermometry and humidity in dry eyes. AB - PURPOSE: To evaluate the relationship of symptoms of dry eye with closed chamber infrared thermometry and humidity measurements. METHODS: The authors studied 54 patients (108 eyes) with dry eye disorders of different grades of clinical severity (mean age 35.75+/-14.37 years), 31 cases (62 eyes) with normal eyes (mean age 33.68+/-14.42 years), and 10 cases (20 eyes) with epidemic conjunctivitis (mean age 33.68+/-14.42 years). The symptoms and the clinical tests (Schirmer-1 test, fluorescein tear break up time, Lissamine green stain; closed chamber infrared thermometry and humidity) were used to establish the diagnosis of dry eyes. The closed chamber humidity difference was used to classify the groups of dry eye. RESULTS: The authors noted no temperature difference from closed to open eye position in dry eyes as compared to 0.10 +/- 0.00 degrees C difference in normal eyes and epidemic conjunctivitis. Four groups of cases were identified by difference in values of humidity: Group 1 = <0.9 relative humidity (RH)% to 1 RH%; Group 2 = >1 RH% to 1.5 RH%; Group 3 = >1.5 RH% to 2 RH %; and Group 4 = >2 RH%. The symptomatology was measured in the eyes using Oxford scale (0-4) and correlated with the humidity groups. The grading of symptoms with the humidity showed a statistically significant relationship (p<0.0001) in each group. The severity of the symptoms showed an increase in frequency and severity from Group 1 to Group 4, which was statistically significant (p<0.0001). The mean sum total of global symptomatology score was statistically significant (p<0.0001): 3.43 +/- 0.31 in Group 1, 4.65+/-0.42 in Group 2, 8.56 +/- 0.78 in Group 3, and 13.35+/-1.21 in Group 4. However, total symptomatology score in epidemic conjunctivitis did not show a statistically significant value (p=0.20). CONCLUSIONS: The closed chamber humidity and thermometry measurements showed statistical correlation in all four groups of dry eyes to total mean symptomatology score (p<0.001) and showed an increase in value with increasing severity of symptoms. However, all individual symptoms fail to show any conclusive relationship. PMID- 15812757 TI - Balloon dilatation for treatment of congenital nasolacrimal duct obstruction. AB - PURPOSE: To evaluate the safety and effectiveness of balloon dacryocystoplasty in the treatment of congenital nasolacrimal duct obstructions. METHODS: Balloon dacryocystoplasty was attempted in 25 eyes of 21 patients. The procedure was performed successfully in 24 eyes of 20 patients, age range 21-72 months. Nineteen eyes had no previous procedure. The mean age of this group was 43.9 months (range 36-72 months). Five eyes had failed probing of lacrimal system. The mean age of this group was 22.2 months (range 21-24 months). The authors performed balloon dacryocystoplasty under endoscopic guidance. Clinical success was defined as complete remission of epiphora within follow-up period of 7-34 months (mean 25.2 months). RESULTS: The authors performed balloon dacryocystoplasty in 24 eyes. The first procedure was successful in 20 of them and the clinic success rate was 83.3%. The technique was repeated in the one eye that recurred and as it ended successfully, the clinic success rate increased to 87.5%. In 17 of the 19 eyes (89.4%) in which balloon dacryocystoplasty was performed primarily, and in 4 of 5 eyes (80%) in which balloon dacryocystoplasty was performed secondarily after unsuccessful probing, the procedure was clinically successful. There was intermittent epiphora in 3 eyes (15%) and these were considered as recurrence. CONCLUSIONS: This experience shows that balloon dilatation is a safe and effective treatment of congenital nasolacrimal duct obstruction as a primary procedure in children over 36 months of age and as a secondary procedure after failure of lacrimal system probing. As a result, balloon dacryocystoplasty can be an alternative treatment in older children and can be preferred to silicone intubation and dacryocystorhinostomy performed after unsuccessful probing. PMID- 15812759 TI - Corneal thickness and functional damage in patients with ocular hypertension. AB - PURPOSE: To correlate functional damage over time detected by standard automated perimetry (SAP) and frequency doubling technology (FDT) with central corneal thickness (CCT) in patients with ocular hypertension (OHT). METHODS: Seventy eight OHT patients underwent CCT measurements, SAP, and FDT (the latter two also after 12 and 18 months). Patients were divided into three equally sized groups of 26 patients each: thin (< 540 microm), normal (540-580 microm), and thick cornea (> 580 microm). The frequency of abnormal FDT and SAP results was analyzed over time (Pearson chi2 test). RESULTS: Six of 26 patients with thin corneas (23.1%) presented an abnormal FDT test at baseline, compared to 1 of 26 (3.8%) in the normal thickness cornea group and 1 of 26 (3.8%) in the thick cornea group. After 12 months, the abnormal FDT tests were as follows, respectively: 9 of 26 (34.6%), 2 of 26 (7.7%), and 2 of 26 (7.7%). For SAP the abnormal results were as follows, respectively: 8 (30.1%), 5 (19.2%), and 2 (7.7%). After 18 months, the abnormal FDT tests were as follows, respectively: 16 (61.5%), 5 (19.2%), and 5 (19.2%). For SAP, the abnormal results were as follows, respectively: 10 (38.5%), 5 (19.2%), and 2 (7.7%). CONCLUSIONS: OHT patients with thinner corneas have a greater risk of developing functional damage over time. PMID- 15812760 TI - Viscocanalostomy with mitomycin-C: a preliminary study. AB - PURPOSE: To compare the results of viscocanalostomy with and without mitomycin-C (MMC). METHODS: Retrospective results of 15 standard viscocanalostomy (VCO) operations (Group 1) were compared with the prospective results of 15 VCO operations performed with intraoperative adjunctive MMC (Group 2). MMC (0.2 mg/mL) was applied over and under the superficial scleral flap for 3 minutes in Group 2 before the deep flap was prepared. Each patient was followed up for at least 1 year, and results of examinations in the first 12 months were used in the statistical comparison of the two groups. Surgical success was defined as intraocular pressure (IOP) < or = 18 mmHg. RESULTS: Preoperative mean intraocular pressures (IOP) in Group 1 and Group 2 were 35.3+/-11.0 and 39.1+/-8.9, respectively. Mean IOP levels at the 12th month were 14.4+/-2.6 and 11.9+/-4.0, respectively, showing a significant decrease in both groups (p<0.001). Postoperative IOP course appeared to be lower in the MMC group, however, the difference was not statistically significant (p=0.554). Complete success rates without medications were 40% in Group 1 and 67% in Group 2. No significant difference was found between the two groups in terms of early and late postoperative complications, pre- and postoperative number of antiglaucoma medications, and surgical success rates at the end of the study period (p>0.05 for all). A significant difference was verified between the two groups of eyes considering the conjunctival bleb types, as low-lying, localized blebs were the most frequent type in Group 1 and thin-walled, avascular blebs were more predominant in the MMC group (p=0.004). CONCLUSIONS: Intraoperative adjunctive MMC use might improve the long-term results of viscocanalostomy by facilitating subconjunctival filtration and might widen the indication range of the technique. PMID- 15812761 TI - Evaluation of two Humphrey perimetry programs: full threshold and SITA standard testing strategy for learning effect. AB - PURPOSE: To compare learning effect of Swedish interactive threshold algorithm (SITA) standard strategy with full threshold testing. METHODS: Thirty-nine medical students with no experience in visual field testing had full threshold (FT) and SITA standard for either right or left eyes. They were chosen in such a way that 20 (Group I) had FT for right and SITA for left eyes and 19 (Group II) had SITA standard for right and FT for left eyes. It was designed to have both strategies on same person whereby eliminating inter-individual variability. Visual field testing was repeated in the same week of the first test on the same subject with the same strategy that was chosen for that eye. RESULTS: The authors found an improvement in mean deviation (MD) and pattern standard deviation (PSD) of first and second testings correspondingly for FT (MD from -3.04 to -2.55; PSD from -2.60 to -2.29) and SITA standard (MD from -2.86 to -2.20; PSD from 2.25 to 2.10) and changes were statistically significant (p<0.05). To analyze learning effect of visual field testings, we calculated percentage change in MD and PSD for full threshold and SITA standard strategy. The percentage changes in visual field parameters were significantly lower in SITA standard strategy testing for MD (p=0.02) and PSD (p=0.01). CONCLUSIONS: This study shows that a learning effect is present for both strategies and SITA standard may have a reduced learning effect compared to FT. PMID- 15812762 TI - Lens epithelial apoptosis and cell proliferation in human age-related cortical cataract. AB - PURPOSE: To probe the presence of apoptosis in the epithelium of human lenses with age-related cortical cataract as well as to assess cell proliferation, a predicted consequence of apoptotic cell death, in this specific cell population. METHODS: DNA fragmentation was assessed using terminal digoxigenin-labeled dUTP nick end labeling (TUNEL) in capsulotomy specimens obtained from patients who underwent either extracapsular cataract extraction for the removal of adult-onset cortical cataract (n=27) or clear lens extraction for the correction of high myopia (n=25). Cell proliferation was assayed in 23 epithelia of cataractous lenses, and 20 epithelia of non-cataractous lenses with the proliferation marker MIB1, a monoclonal antibody against the nuclear antigen Ki-67 that is detected throughout the cell cycle but is absent in the resting (G0) cell. RESULTS: TUNEL staining was observed in 25 (92.6%) specimens of cataractous lenses, whereas cells undergoing apoptosis were identified in 2 (8%) of the epithelia from non cataractous lenses. Only two MIB1-positive samples were detected, one of which was a capsule obtained during intracapsular cataract extraction. CONCLUSIONS: The epithelium of human lenses with cortical cataract undergoes low rate apoptotic death. This limited epithelial apoptosis is unlikely to result in any significant cell density decrease since epithelial gaps are likely to be replaced by cell proliferation at the germinative zone of the anterior lens capsule. Nevertheless, the accumulation of small-scale epithelial losses during lifetime may induce alterations in lens fiber formation and homeostasis and result in loss of lens transparency. PMID- 15812763 TI - Uncomplicated phacoemulsification and first-day review: the patient's perspective. AB - PURPOSE: To evaluate patients'opinions regarding first-day review following uncomplicated phacoemulsification. METHODS: A prospective questionnaire-based survey was conducted on consecutive patients at the time of listing for day-case phacoemulsification. Data collected included demographic details, proposed anaesthetic technique, first or second eye surgery and postcode. Statistical analysis was undertaken with the unpaired t-test, the chi2-test with Yates' correction for small numbers or Fisher's exact test, and Kendal's rank correlation, as appropriate. RESULTS: 100 patients (59% female; mean age 74.2 (12.6) years) were recruited. 87% preferred to return to hospital for their first day review. This result was not significantly affected by sex, proposed anaesthetic technique, first eye surgery or postcode area. Only patients < 65 years old preferred not to return (p<0.02). CONCLUSIONS: In this population, patients preferred to return for first-day review following uncomplicated phacoemulsification. There may be some variance in the opinions of surgeons and patients regarding this issue, and the patient's preferences should be taken into account to provide optimum quality of care. PMID- 15812764 TI - Review of sterile, postoperative, anterior segment inflammation following cataract extraction and intraocular lens implantation. AB - PURPOSE: To evaluate unexpected, acute intraocular anterior segment inflammation following uneventful cataract extraction by phacoemulsification and foldable posterior chamber intraocular lens (PCIOL) implantation. METHODS: This retrospective study evaluated five cases of postoperative inflammation that occurred after cataract extraction with foldable PCIOL implantation. Medical records were reviewed to analyze the unexplained postoperative inflammation. RESULTS: The five patients who developed inflammatory activity during the postoperative period responded well to corticosteroid treatment. Toxic maculopathy developed in one patient after aggressive antibacterial therapy. Vitrectomy was performed for one patient with prolonged vitreal inflammatory activity. CONCLUSIONS: Noninfectious endophthalmitis developing upon surgery may be caused by a multifactorial process or an interindividual variable response to a common factor as a hypersensitivity reaction. It should be remembered in inflammatory cases after surgery in order to prevent the toxic, irreversible side effects of bacterial endophthalmitis treatment. PMID- 15812765 TI - Effect of prophylactic brimonidine on bleeding complications after cataract surgery. AB - PURPOSE: To evaluate the effect of prophylactic brimonidine on bleeding complications after cataract surgery. METHODS: The authors performed a prospective, double-masked, two-surgeon study of 137 patients (137 eyes) who underwent phacoemulsification and intraocular lens implantation with or without prophylactic brimonidine before cataract surgery. The authors also compared the effect of brimonidine among patients with systemic diseases such as diabetes mellitus (types I and II), hypertension, and anticoagulant or antiplatelet treatment. RESULTS: Subconjunctival hemorrhage was observed in 73.70% of the patients not treated with brimonidine before surgery and in only 23.75% of the patients who were given prophylactic brimonidine (p<0.001, chi2). The grade of hemorrhage was also statistically significant (p<0.001, Mann-Whitney). No statistically significant difference with regard to the presence of hemorrhage in diabetic patients or in the anticoagulant or antiplatelet treatment group was observed. However, a statistically significant difference (p<0.027, chi2) was found between hypertensive patients treated and not treated with prophylactic brimonidine before cataract surgery. CONCLUSIONS: This study suggests that brimonidine administered before cataract surgery may significantly reduce subconjunctival hemorrhage in the general population. It has been shown to be beneficial in hypertensive patients. A strong statistical trend, but not significance has been found in diabetic patients or in patients treated with antiplatelet or anticoagulant drugs, but further studies are needed to reach conclusive results. PMID- 15812766 TI - A short duration transient ischemia induces apoptosis in retinal layers: an experimental study in rabbits. AB - PURPOSE: To investigate retinal cell apoptosis in an experimental transient, short duration ocular ischemia model. METHODS: An experimental ischemia model, which simulates creating temporary high intraocular pressure to control intraocular bleeding during pars plana vitrectomy, was set up. Rabbits were randomly divided into three groups. Group 1 was the control group. In Group 2, intraocular pressure was increased to 97 mmHg for 5 minutes. In Group 3, intraocular pressure was increased to 97 mmHg for 10 minutes. After 24 hours, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling assay was used to detect retinal apoptosis in rabbit eyes. Only nuclear staining in retinal cells was counted. RESULTS: Groups with 5 minutes and 10 minutes of ischemia showed significantly higher amount of ganglion cell layer apoptosis when compared with the control group (p<0.05). Light microscopy and standard hematoxylin-eosin did not show any significant damage in the retina cells. CONCLUSIONS: Apoptotic cell death in the retinal cell layers occurs in temporary ischemia-reperfusion as early as 5 and 10 minutes duration. PMID- 15812767 TI - Comparing measurements of retinal nerve fiber layer thickness obtained on scanning laser polarimetry with fixed and variable corneal compensator. AB - PURPOSE: To compare retinal nerve fiber layer (RNFL) thickness measurements obtained on scanning laser polarimetry (SLP) with commercially available instruments coupled with fixed (FCC) and variable corneal compensator (VCC). METHODS: Forty-two eyes of 42 patients underwent a complete ophthalmologic evaluation and achromatic automated perimetry (24-2 program, SITA standard strategy). Nineteen eyes were healthy (average mean deviation: -0.12 dB +/- 2.26) and 23 glaucomatous (average mean deviation: -4.92 dB +/- 6.49). All patients underwent SLP with both FCC and VCC. Adequate compensation of corneal birefringence on FCC-SLP was checked acquiring macular retardation map (MRM). RNFL thickness was evaluated considering superior and inferior maximum (SM, IM), average thickness and ellipse average (AT, EA), and superior and inferior average (SA, IA). Mean values (+/-SD) for each parameter measured by the two polarimeters were compared and linear regression calculated. The ability of each parameter to discriminate between normal and glaucomatous eyes was evaluated on both polarimeters calculating area under ROC curve. RESULTS: A significant linear correlation for all parameters was noted (r range: 0.65-0.78). VCC produced slightly higher thickness values than FCC, both in normal and glaucomatous eyes. On both polarimeters, area under ROC curve for all parameters discriminated adequately healthy from glaucomatous eyes (range: 0.68-0.81). CONCLUSIONS: In a highly comparable and selected group of normal and glaucomatous eyes, FCC-SLP and VCC-SLP showed considerable concordance in measuring peripapillary RNFL thickness, both for sectorial and global parameters. Proper corneal birefringence compensation provided separation of normal from glaucomatous eyes on both polarimeters. PMID- 15812768 TI - Retinal nerve fiber layer measured by Heidelberg retina tomograph and nerve fiber analyzer. AB - PURPOSE: To compare retinal nerve fiber layer (RNFL) thickness measured by Heidelberg retina tomograph (HRT) and nerve fiber analyzer (GDx). METHODS: Twenty eyes of 20 consecutive healthy subjects were recruited for this study. Each subject had a normal visual field and a normal optic nerve head, which was assessed by slit-lamp biomicroscopy using a 90 degrees lens. Using the HRT and GDx, RNFL measurement was calculated as for software vs 2.01 and vs 1.0.14, respectively. Retinal nerve fiber layer thickness was evaluated for the entire annulus surface every 5 degrees degrees. RNFL was assessed by HRT and GDx. HRT RNFL measurement was calculated at 0 microm from the edge, while GDx RNFL measurement at 1.75 disc diameter as for software. The difference between the highest points and the deepest points was calculated and compared. Furthermore, because of the possibility of different scales in the two systems, the following ratio was calculated: superior/inferior, superior/temporal, superior/nasal, inferior/temporal, and inferior/nasal. RESULTS: When the entire RNFL thickness was considered, a significant (p<0.001) difference was found between the HRT and GDx measurements. A difference of 200 microm was found between the highest and the deepest HRT points while a difference of 40 microm was found between the highest and the deepest GDx points. CONCLUSIONS: HRT and GDx RNFL measurements were statistically different in each sector. However, ratio parameters showed no difference between the obtained values except for superior/temporal ratio and inferior/temporal ratio. PMID- 15812769 TI - Early age-related maculopathy and risk factors of cardiovascular disease in middle-aged Lithuanian urban population. AB - PURPOSE: To assess the prevalence of age-related maculopathy (ARM) in a middle aged urban population and the relationship between ARM and the main risk factors of cardiovascular disease (CVD). METHODS: The survey according to the WHO MONICA study protocol was carried out in Kaunas city, Lithuania, from 2001 to 2002. A total of 1403 persons aged 35 to 64 years were screened (626 men and 777 women: response rate 62.4%). Ophthalmologic investigation was performed for 1337 respondents (594 men and 743 women). Analysis of the relationship between ARM and risk factors of CVD was performed in case-control subdivision matching for sex, age, and education level. RESULTS: Early ARM was diagnosed in 7.4% of men and 5.4% of women. Rate of current smoking in case and control groups did not differ in men but in case group of women it was greater than in control group. Mean systolic and diastolic blood pressure and body mass index (BMI) in male case group and mean fasting blood glucose concentration in female case group were higher than in corresponding control groups. Frequency of diastolic hypertension (diastolic blood pressure > or = 90 mmHg) and obesity (BMI > or = 30 kg/m2) in male case group was higher than in control group. ARM was not associated with cholesterol and triglyceride levels in men and women. CONCLUSIONS: Early ARM in middle-aged Lithuanian urban population was associated with current smoking in women but not in men; it was associated with diastolic blood pressure and BMI in men and with fasting glucose level in women. PMID- 15812770 TI - Idiopathic sclerosing inflammation of the orbit: a case of steroid-responsive disease in a patient with auto-immune hemolytic anemia. AB - PURPOSE: Idiopathic sclerosing orbital inflammation (ISOI) has been categorized by some authors as a unique clinicopathologic entity, separate from the heterogeneous group of disorders known collectively as nonspecific orbital inflammation. Histologic similarity and clinical association with other fibrosclerosing conditions has been shown. The authors present a case of ISOI in conjunction with autoimmune hemolytic anemia. METHODS: A 59-year-old woman with a history of hemolytic anemia had left upper lid swelling, periocular pain, proptosis, and restriction of ocular motility. Magnetic resonance imaging (MRI) showed a homogeneously enhancing lateral orbital mass. Biopsy revealed dense fibrous connective tissue with a paucicellular infiltrate, consistent with ISOI, and treatment with prednisolone 60 mg/day was instituted. RESULTS: At 4-week review, the proptosis had settled and the patient regained full range of extraocular movements. At 14 months, the response was sustained and repeat MRI showed a 70% reduction in size of the mass. CONCLUSIONS: The first known case of ISOI and hemolytic anemia is presented. Despite dense fibrosis histologically, steroid responsiveness can be encountered in cases of ISOI. An early, aggressive approach to management is recommended, and corticosteroids should be considered as a treatment option. PMID- 15812771 TI - Ectropion of the lacrimal point: the shoelace technique. AB - PURPOSE: To describe an alternative technique to repair the ectropion of the lacrimal point, either alone or combined with an ectropion related to outer angle laxity. METHODS/RESULTS: After diamond shape resection of the conjunctiva and the retractors, sutures are placed with each bridle interlacing on the posterior portion of the eyelid below the lacrimal point in a shoelace fashion. CONCLUSIONS: The technique combines treatment of hyperlaxity of several anatomic structures in a single operation and has the advantage of reinforcing the Horner muscle, which is essential for the cure of this type of ectropion. PMID- 15812772 TI - Corneal graft rejection after penetrating keratoplasty for keratoconus in Turner's syndrome. AB - PURPOSE: To report a patient with Turner's syndrome who developed graft rejection after penetrating keratoplasty (PK) for keratoconus and to review the ophthalmic literature on the association between keratoconus and Turner's syndrome. METHODS: A woman with bilateral keratoconus and Turner's syndrome (45,XO) was referred for progressive visual loss in the right eye. Best-corrected visual acuity was 20/400 in the right eye. Slit-lamp examination revealed corneal thinning with ectatic protrusion of the central cornea and Vogt's striae in the right eye. The patient underwent PK in the right eye in January 2001. She developed graft rejection in April 2003 and visual acuity dropped to hand motion. After treatment with topical and systemic steroids and systemic cyclosporine A, visual acuity recovered to 20/80 in July 2003. RESULTS: The authors know of only three other reported patients (six eyes) with keratoconus in Turner's syndrome. Five eyes underwent PK with good visual rehabilitation, but one developed immunologic graft rejection 7 years after surgery. On the whole, considering the current report and the other cases described in the literature, graft rejection occurred in 2 out of 6 eyes (33.3%). The graft survival rate was 80% after 2 years and 40% after 7 years. CONCLUSIONS: The results suggest that grafts for keratoconus in patients with Turner's syndrome might have an increased risk of immunologic rejection. Corneal grafts in Turner's syndrome need to be monitored closely. Early detection of graft rejection and aggressive treatment with topical and systemic steroids and systemic cyclosporine A can save the graft and restore useful vision. PMID- 15812773 TI - Amniotic membrane transplantation associated with conjunctival peritomy in the management of Mooren's ulcer: a case report. AB - PURPOSE: To report the association of conjunctival peritomy with amniotic membrane transplantation (AMT) at the limbus with the exclusion of the central cornea in order to preserve visual function in one case of bilateral Mooren's ulcer. METHODS: A 36-year-old man with bilateral Mooren's ulcer was unresponsive to conventional therapy. Surgical procedure was performed on his right eye, at impending risk of corneal perforation. A 20 x 20 mm piece of amniotic membrane (AM) was prepared by performing a central hole of 7.5 mm diameter with a manual trephine. A 360 degrees conjunctival peritomy was performed and the AM was placed with the epithelium side facing up and the central hole was sutured on the paracentral cornea. RESULTS: Two weeks after surgery, while the right eye showed improvement of signs and symptoms and unchanged best-corrected visual acuity (BCVA), the left eye showed a peripheral corneal perforation with prolapsed iris that required conjunctival flap. At 7 months of follow-up, the right eye showed no ocular inflammation, a reduction of the lipid-like peripheral corneal infiltrates, an increased stromal thickness, and an unchanged BCVA. The progression of corneal thinning in the left eye led the authors to perform AMT (as described) in the left eye as well. Five months after the AMT in the left eye, neither eye shows signs of disease progression, and neither requires further therapy. CONCLUSIONS: Conjunctival peritomy associated with AMT may be an alternative surgical approach in the management of Mooren's ulcers to control the inflammation and the progression of disease. PMID- 15812774 TI - Iris atrophy, serous detachment of the ciliary body, and ocular hypotony in chronic phase of Vogt-Koyanagi-Harada disease. AB - PURPOSE: To describe iris atrophy, serous detachment of the ciliary body, and ocular hypotony in a patient with chronic phase of Vogt-Koyanagi-Harada (VKH) disease. METHODS: Ocular examination and follow-up including digital infrared transillumination imaging of the iris was done in a 52-year-old woman with chronic phase of VKH disease. RESULTS: Infrared transillumination imaging showed extensive atrophy of the iris stroma and occasional pigment clumps both in the pupillary and ciliary zones of the iris, and detachment of the ciliary body in both eyes. Conventional transpupillary transillumination using white light showed only minute patchy atrophy of the pigment epithelium in the pupillary zone. Treatment did not normalize bilateral shallow retinal detachment of the posterior pole, serous detachment of the ciliary body, or severe ocular hypotony. CONCLUSIONS: Severe atrophy of the iris stroma, retinal detachment of the posterior pole, serous detachment of the ciliary body, and ocular hypotony may occur in chronic phase of VKH disease. PMID- 15812775 TI - Bilateral serous macular detachment as a presenting feature of acute lymphoblastic leukemia. AB - PURPOSE: To report a case of bilateral serous maculopathy as an initial sign of acute lymphoblastic leukaemia in children. METHODS/RESULTS: A 13-year-old girl, who presented with symptoms of visual blurring, was found to have a bilateral serous maculopathy. Haematological abnormalities (thrombocytopenia with a mild lymphocytosis) prompted further investigation. A bone marrow aspirate revealed the presence of leukemic blasts and a diagnosis of acute lymphoblastic leukaemia was made. Her maculopathy completely resolved following systemic chemotherapy. CONCLUSIONS: Prompt recognition of disease led to early systemic treatment and restoration of visual function. PMID- 15812776 TI - Acute retinal necrosis presenting as central retinal artery occlusion with cilioretinal sparing. AB - PURPOSE: To report a case of acute retinal necrosis presenting as central retinal artery occlusion with cilioretinal sparing. METHODS: Single interventional case report. The findings of the ophthalmic examination, MRI, blood parameters, biopsy results and clinical course are reported. RESULTS: A forty two year old gentleman reporting sudden loss of sight, ophthalmic examination revealing uveitis, central retinal artery occlusion with cilioretinal sparing and peripheral necrotizing retinitis. CONCLUSIONS: Central retinal artery occlusion can be an early feature of acute retinal necrosis (ARN). PMID- 15812778 TI - Saturation transfer and chemical exchange measurements of the stereochemical drift occurring during the Wittig reaction. AB - The Wittig reaction of butylidenetriphenylphosphorane with benzaldehyde using LiHMDS as base in THF was studied. The stereochemical drift (different ratio obtained in alkenes versus oxaphosphetane intermediates) was followed by low temperature 1D NMR techniques. A retro-Wittig reaction is demonstrated using 13C and 31P saturation transfer experiments and homonuclear DPFGSE-ROE techniques. PMID- 15812777 TI - Macular edema and visual loss after macular pucker surgery with ICG-assisted internal limiting membrane peeling. AB - PURPOSE: To describe the occurrence of massive macular edema and visual loss after indocyanine green-assisted (ICG) macular pucker surgery. METHODS/RESULTS: A 74 years old female presented with a macular pucker and a hypertrophy of the retinal pigment epithelium (RPE) in her left eye. The preoperative visual acuity (VA) was 20/100. Surgery consisted of cataract extraction, lens implantation and standard pars plana vitrectomy with peeling of epiretinal tissue followed by the removal of the internal limiting membrane (ILM) remnants stained using a 0.05% ICG solution. One day after surgery, VA was counting fingers. There was an extensive macular edema and retinal thickening with hyperfluorescence during fluorescein angiography and pronounced autofluorescence using ICG filters. During follow up, the macular edema resolved completely, but VA decreased to 20/800 at six months postoperatively. There was a central scotoma and unstable fixation seen during microperimetry. DISCUSSION: This case report indicates that ICG might come into contact with bare retina if injected following removal of epiretinal membranes. Whether the observed RPE hypertrophy might have contributed to the pathogenesis of the adverse effect described remains hypothetical. PMID- 15812780 TI - Optimal virtual orbitals to relax wave functions built up with transferred extremely localized molecular orbitals. AB - Extremely localized molecular orbitals (ELMOs), namely orbitals strictly localized on molecular fragments, are easily transferable from one molecule to another one. Hence, they provide a natural way to set up the electronic structure of large molecules using a data base of orbitals obtained from model molecules. However, this procedure obviously increases the energy with respect to a traditional MO calculation. To gain accuracy, it is important to introduce a partial electron delocalization. This can be carried out by defining proper optimal virtual orbitals that supply an efficient set for nonorthogonal configurations to be employed in VB-like expansions. PMID- 15812779 TI - New AMBER force field parameters of heme iron for cytochrome P450s determined by quantum chemical calculations of simplified models. AB - The heme protein, cytochrome P450, is an oxidoreductase that plays an important role in drug metabolism. To model P450s using molecular mechanics methods and classical molecular dynamics simulations, force field parameters and atomic charges are required. Because these parameters are generally obtained by quantum chemical methods, an appropriate simplified model for the iron-porphyrin system was needed. In this study, two models with a five-coordinated Fe(III) mimicking the sextet spin state of P450s are proposed, which are optimized by semiempirical and ab initio unrestricted Hartree-Fock methods. The results produced using the simpler of the two models were similar to those of the more complex model; therefore, the more simplified model of P450 can be used without a loss of accuracy. Furthermore, several quantum chemical calculations were carried out on the simpler model to investigate which method was most suitable for iron porphyrin systems. The results calculated by hybrid density functional theory (DFT), with the MIDI basis set for iron, reproduced the three-dimensional structures determined by X-ray diffraction and extended X-ray absorption fine structure experiments. From these results, atomic charges and force-field parameters for molecular mechanics and molecular dynamics calculations were obtained. PMID- 15812782 TI - Searching sequence space: two different approaches to dihydrofolate reductase catalysis. AB - There are numerous examples of proteins that catalyze the same reaction while possessing different structures. This review focuses on two dihydrofolate reductases (DHFRs) that have disparate structures and discusses how the catalytic strategies of these two DHFRs are driven by their respective scaffolds. The two proteins are E. coli chromosomal DHFR (Ec DHFR) and a type II R-plasmid-encoded DHFR, typified by R67 DHFR. The former has been described as a very well evolved enzyme with an efficiency of 0.15, while the latter has been suggested to be a model for a "primitive" enzyme that has not yet been optimized by evolution. This comparison underlines what is important to catalysis in these two enzymes and concurrently highlights fundamental issues in enzyme catalysis. PMID- 15812783 TI - Convenient access both to highly antimalaria-active 10-arylaminoartemisinins, and to 10-alkyl ethers including artemether, arteether, and artelinate. AB - An economical phase-transfer method is used to prepare 10-arylaminoartemisinins from DHA and arylamines, and artemether, arteether, and artelinate from the corresponding alcohols. In vivo sc screens against Plasmodium berghei and P. yoelii in mice reveal that the p-fluorophenylamino derivative 5 g is some 13 and 70 times, respectively, more active than artesunate; this reflects the very high sc activity of 10-alkylaminoartemisinins. However, through the po route, the compounds are less active than the alkylaminoartemisinins, but still approximately equipotent with artesunate. PMID- 15812784 TI - Generation of novel landomycins M and O through targeted gene disruption. AB - Two genes from Streptomyces cyanogenous S136 that encode the reductase LanZ4 and the hydroxylase LanZ5, which are involved in landomycin A biosynthesis, were characterized by targeted gene inactivation. Analyses of the corresponding mutants as well as complementation experiments have allowed us to show that LanZ4 and LanZ5 are responsible for the unique C-11-hydroxylation that occurs during landomycin biosynthesis. Compounds accumulated by the lanZ4/Z5 mutants are the previously described landomycin F and the new landomycins M and O. PMID- 15812785 TI - Potentiometric Detection of Single Nucleotide Polymorphism by Using a Genetic Field-effect transistor. AB - Potentiometric measurement of allele-specific oligonucleotide hybridization based on the principle of detection of charge-density change at the surface of a gate insulator by using of a genetic field-effect transistor has been demonstrated. Since DNA molecules are negatively charged in aqueous solution, a hybridization event at the gate surface leads to a charge-density change in the channel of the FET and can be directly transduced into an electrical signal without any labeling of target DNA molecules. One of the unique features of our method is to utilize DNA binders such as intercalators as charged species for double-stranded DNA after hybridization, since these are ionized and carry positive charges in aqueous solution. Single-base mismatch of the target DNA could be successfully detected both with the wild-type and with the mutant genetic FETs by controlling the hybridization temperatures and introducing Hoechst 33258 as DNA binder. The genetic FET platform is suitable as a simple, accurate, and inexpensive system for SNP typing in clinical diagnostics. PMID- 15812788 TI - Structural chemoproteomics and drug discovery. AB - Our laboratories have developed several technologies to accelerate drug discovery process on the basis of structural chemoproteomics. They include SPS technology for the efficient determination of protein structures, SCP technology for the rapid lead generation and SDF technology for the productive lead optimization. Using these technologies, we could determine many 3D structures of target proteins bound with biologically active chemicals including the structure of phosphodiesterase 5/Viagra complex and obtain highly potent compounds in animal models of obesity, diabetes, cancer and inflammation. In this paper, we will discuss concepts and applications of structural chemoproteomics for drug discovery. PMID- 15812786 TI - Inversion of stereoselectivity by applying mutants of the hydroxynitrile lyase from Manihot esculenta. AB - The influence of Trp128-substituted mutants of the hydroxynitrile lyase from Manihot esculenta (MeHNL) on the stereoselectivity of MeHNL-catalyzed HCN additions to aldehydes with stereogenic centers, which yield the corresponding cyanohydrins, is described. In rac-2-phenylpropionaldehyde (rac-1) reactions, wild-type (wtMeHNL) and all MeHNL Trp128 mutants are highly (S)-selective toward the (R) enantiomer of rac-1; this results exclusively in (2S,3R)-cyanohydrin ((2S,3R)-2) with > or =96 % de. The (S) enantiomer of rac-1, however, only reacts (S)-selectively with wtMeHNL to give (2S,3S)-2 with 80 % de, whereas with Trp128 mutants, (R) selectivity increases with decreasing size of the amino acids exchanged. The MeHNL W128A mutant is exclusively (R)-selective, resulting in (2R,3S)-2 with 86 % de. The reaction behavior of rac-phenylbutyraldehyde (rac-5) is comparable with rac-1, which also inverts the stereoselectivity from (S) to (R) when the enzyme is exchanged from wtMeHNL to the W128A mutant. Stereogenic centers not adjacent to the aldehyde group, as in 7 and 9, do not influence the stereoselectivity of MeHNL catalysis, and (S) selectivity is observed in all cases. Stereoselectivity and inversion of stereoselectivity of MeHNL Trp128 mutant-catalyzed cyanohydrin formation can be explained and rationalized with crystal-structure-based molecular modeling. PMID- 15812789 TI - Theoretical study on the reaction mechanism of the methyl radical with nitrogen oxides. AB - The radical-molecule reaction mechanism of CH3 with NOx (x = 1, 2) has been explored theoretically at the B3LYP/6-311Gd,p and MC-QCISD (single-point) levels of theory. For the singlet potential energy surface (PES) of the CH3 + NO2 reaction, it is found that the carbon to middle nitrogen attack between CH3 and NO2 can form energy-rich adduct a (H3CNO2) with no barrier followed by isomerization to b1 (CH3ONO-trans), which can easily convert to b2 (CH3ONO-cis). Subsequently, starting from b (b1, b2), the most feasible pathway is the direct N O bond cleavage of b (b1, b2) leading to P1 (CH3O + NO) or the 1,3-H-shift and N O bond rupture of b1 to form P2 (CH2O + HNO), both of which may have comparable contribution to the reaction CH3 + NO2. Much less competitively, b2 can take a concerted H-shift and N-O bond cleavage to form product P3 (CH2O + HON). Because the intermediates and transition states involved in the above three channels are all lower than the reactants in energy, the CH3 + NO2 reaction is expected to be rapid, as is consistent with the experimental measurement in quality. For the singlet PES of the CH3 + NO reaction, the major product is found to be P1 (HCN + H2O), whereas the minor products are P2 (HNCO + H2) and P3 (HNC +H2O). The CH3 + NO reaction is predicted to be only of significance at high temperatures because the transition states involved in the most feasible pathways lie almost above the reactants. Compared with the singlet pathways, the triplet pathways may have less contributions to both reactions. The present study may be helpful for further experimental investigation of the title reactions. PMID- 15812790 TI - Effect of the metal-assisted assembling mode on the redox states of hexaazatriphenylene hexacarbonitrile. PMID- 15812791 TI - Uniform, axial-orientation alignment of one-dimensional single-crystal silicon nanostructure arrays. PMID- 15812795 TI - Amino acids--valuable organocatalysts in carbohydrate synthesis. PMID- 15812796 TI - Chiral amplification in the transcription of supramolecular helicity into a polymer backbone. PMID- 15812798 TI - Anaerobic treatment of complex chemical wastewater in a sequencing batch biofilm reactor: process optimization and evaluation of factor interactions using the Taguchi dynamic DOE methodology. AB - The Taguchi robust experimental design (DOE) methodology has been applied on a dynamic anaerobic process treating complex wastewater by an anaerobic sequencing batch biofilm reactor (AnSBBR). For optimizing the process as well as to evaluate the influence of different factors on the process, the uncontrollable (noise) factors have been considered. The Taguchi methodology adopting dynamic approach is the first of its kind for studying anaerobic process evaluation and process optimization. The designed experimental methodology consisted of four phases- planning, conducting, analysis, and validation connected sequence-wise to achieve the overall optimization. In the experimental design, five controllable factors, i.e., organic loading rate (OLR), inlet pH, biodegradability (BOD/COD ratio), temperature, and sulfate concentration, along with the two uncontrollable (noise) factors, volatile fatty acids (VFA) and alkalinity at two levels were considered for optimization of the anae robic system. Thirty-two anaerobic experiments were conducted with a different combination of factors and the results obtained in terms of substrate degradation rates were processed in Qualitek-4 software to study the main effect of individual factors, interaction between the individual factors, and signal-to-noise (S/N) ratio analysis. Attempts were also made to achieve optimum conditions. Studies on the influence of individual factors on process performance revealed the intensive effect of OLR. In multiple factor interaction studies, biodegradability with other factors, such as temperature, pH, and sulfate have shown maximum influence over the process performance. The optimum conditions for the efficient performance of the anaerobic system in treating complex wastewater by considering dynamic (noise) factors obtained are higher organic loading rate of 3.5 Kg COD/m3 day, neutral pH with high biodegradability (BOD/COD ratio of 0.5), along with mesophilic temperature range (40 degrees C), and low sulfate concentration (700 mg/L). The optimization resulted in enhanced anaerobic performance (56.7%) from a substrate degradation rate (SDR) of 1.99 to 3.13 Kg COD/m3 day. Considering the obtained optimum factors, further validation experiments were carried out, which showed enhanced process performance (3.04 Kg COD/m3-day from 1.99 Kg COD/m3 day) accounting for 52.13% improvement with the optimized process conditions. The proposed method facilitated a systematic mathematical approach to understand the complex multi species manifested anaerobic process treating complex chemical wastewater by considering the uncontrollable factors. PMID- 15812799 TI - Stoichiometric limitations in assembly of active recombinant retrovirus. AB - Although recombinant retroviruses are widely used in gene therapy and as gene transfer vehicles for basic biological studies, their titers are very low as compared to other recombinant viral systems, e.g., adenovirus. We investigated the rate-limiting steps in production of LacZ-encoding ecotropic (CRE BAG 2) and amphotropic (Psi-CRIP) retrovirus. We found that ecotropic retrovirus producer cells produced a large number of inactive viral particles because they were severely limited by the amount of mRNA that was packaged into viral capsids. Introduction of the gene for green fluorescence protein (GFP) increased retroviral titers 40-fold, without affecting the viral matrix protein, p30, or the activity of reverse transcriptase. Surprisingly, while transfer of GFP gene increased retrovirus production, beta-gal activity and X-gal titer decreased significantly. Quantitative real-time polymerase chain reaction (PCR) showed that although producer cells synthesized similar amounts of both mRNAs, retroviral supernatants contained significantly lower amount of LacZ mRNA, possibly due to competition between LacZ and GFP mRNAs for encapsidation into virions. In contrast to ecotropic producers, introduction of GFP gene copies into amphotropic producers resulted in a moderate twofold increase in retrovirus production. However, delivery of genes encoding for the viral proteins gp70 and p30 increased virus production by fivefold, suggesting that amphotropic producers may also be limited by synthesis of structural viral proteins. Our data show that in addition to the amount of viral genome or proteins, assembly of viral components into active viral particles may limit production of high titer retroviral preparations. PMID- 15812800 TI - A novel purification strategy for retrovirus gene therapy vectors using heparin affinity chromatography. AB - Membrane separation and chromatographic technologies are regarded as an attractive alternative to conventional academic small-scale ultracentrifugation procedures used for retrovirus purification. However, despite the increasing demands for purified retroviral vector preparations, new chromatography adsorbents with high specificity for the virus have not been reported. Heparin affinity chromatography is presented here as a novel convenient tool for retrovirus purification. The ability of bioactive retroviral particles to specifically bind to heparin ligands immobilized on a chromatographic gel is shown. A purification factor of 63 with a recovery of 61% of functional retroparticles was achieved using this single step. Tentacle heparin affinity supports captured retroviral particles more efficiently than conventional heparin affinity chromatography supports with which a lower recovery was obtained (18%). Intact, infective retroviral particles were recovered by elution with low salt concentrations (350 mM NaCl). Mild conditions for retrovirus elution from chromatographic columns are required to preserve virus infectivity. VSV-G pseudotyped retroviruses have shown to be very sensitive to high ionic strength, losing 50% of their activity and showing membrane damage after a short exposure to 1M NaCl. We also report a complete scaleable downstream processing scheme for the purification of MoMLV-derived vectors that involves sequential microfiltration and ultra/diafiltration steps for virus clarification and concentration respectively, followed by fractionation by heparin affinity chromatography and final polishing by size-exclusion chromatography. Overall, by using this strategy, a 38% yield of infective particles can be achieved with a final purification factor of 2,000. PMID- 15812801 TI - Metabolite profiling for analysis of yeast stress response during very high gravity ethanol fermentations. AB - A laboratory strain and an industrial strain of Saccharomyces cerevisiae were grown at high substrate concentration, so-called very high gravity (VHG) fermentation. Simultaneous saccharification and fermentation (SSF) was applied in a batch process using 280 g/L maltodextrin as carbon source. It was shown that known ethanol and osmotic stress responses such as decreased growth rate, lower viability, higher energy consumption, and intracellular trehalose accumulation occur in VHG SSF for both strains when compared with standard laboratory medium (20 g/L glucose). The laboratory strain was the most affected. GC-MS metabolite profiling was applied for assessing the yeast stress response influence on cellular metabolism. It was found that metabolite profiles originating from different strains and/or fermentation conditions were unique and could be distinguished with the help of multivariate data analysis. Several differences in the metabolic responses to stressing conditions were revealed, particularly the increased energy consumption of stressed cells was also reflected in increased intracellular concentrations of pyruvate and related metabolites. PMID- 15812802 TI - Separation of monoclonal antibody alemtuzumab monomer and dimers using ultrafiltration. AB - This article examines the feasibility of using ultrafiltration to separate the monomer of the monoclonal antibody alemtuzumab (Campath or Campath-1H) from a mixture of dimer and higher-order oligomers (collectively called "dimers" here). Using parameter scanning ultrafiltration, we initially assessed the suitability of the following membranes: 100 kDa and 300 kDa polyethersulfone (PES) membranes, and a 100 kDa polyvinylidene fluoride (PVDF) membrane. A detailed study was then carried out to examine the effects of operating conditions (such as solution pH, ionic strength, stirring speed, and permeate flux) on the separation of the monomer from the dimers using 300 kDa PES and 100 kDa PVDF membranes. Results of the experiments carried out in the carrier phase ultrafiltration (CPUF) mode indicate that the size-based protein-protein separation critically depends on the membrane used as well as the system hydrodynamics. The separation of the monoclonal antibody monomer and dimers using 100 kDa PVDF membranes in the diafiltration mode was also examined. Experimental results demonstrate that under suitable conditions, it is feasible to obtain the alemtuzumab monomer with a purity of more than 93% and a yield of more than 85% (from a mixture of 75% monomer and 25% dimers, which is the typical composition obtained after affinity chromatography). Simulation study indicates that this could be further improved to a purity of more than 96% and a monomer yield of more than 96% by increasing the selectivity of separation or by employing a two-stage diafiltration process. PMID- 15812803 TI - NOx removal from flue gas by an integrated physicochemical absorption and biological denitrification process. AB - An integrated physicochemical and biological technique for NO(x) removal from flue gas, the so-called BioDeNO(x) process, combines the principles of wet absorption of NO in an aqueous Fe(II)EDTA(2-) solution with biological reduction of the sorbed NO in a bioreactor. The biological reduction of NO to di-nitrogen gas (N(2)) takes place under thermophilic conditions (55 degrees C). This study demonstrates the technical feasibility of this BioDeNO(x) concept in a bench scale installation with a continuous flue gas flow of 650 l.h(-1) (70-500 ppm NO; 0.8-3.3% O(2)). Stable NO removal with an efficiency of at least 70% was obtained in case the artificial flue gas contained 300 ppm NO and 1% O(2) when the bioreactor was inoculated with a denitrifying sludge. An increase of the O(2) concentration of only 0.3% resulted in a rapid elevation of the redox potential (ORP) in the bioreactor, accompanied by a drastic decline of the NO removal efficiency. This was not due to a limitation or inhibition of the NO reduction, but to a limited biological iron reduction capacity. The latter leads to a depletion of the NO absorption capacity of the scrubber liquor, and thus to a poor NO removal efficiency. Bio-augmentation of the reactor mixed liquor with an anaerobic granular sludge with a high Fe(III) reduction capacity successfully improved the bioreactor efficiency and enabled to treat a flue gas containing at least 3.3% O(2) and 500 ppm NO with an NO removal efficiency of over 80%. The ORP in the bioreactor was found to be a proper parameter for the control of the ethanol supply, needed as electron donor for the biological regeneration process. The NO removal efficiency as well as the Fe(III)EDTA(-) reduction rate were found to decline at ORP values higher than -140 mV (pH 7.0). For stable BioDeNO(x) operation, the supply of electron donor (ethanol) can be used to control the ORP below that critical value. PMID- 15812805 TI - Cell cycle analysis of Taxus suspension cultures at the single cell level as an indicator of culture heterogeneity. AB - Single cell growth and division was measured via flow cytometry in order to characterize the metabolic variability of Taxus cuspidata suspension cultures, which produce the valuable secondary metabolite Taxol. Good agreement was observed between the cell cycle distribution and biomass accumulation over the batch culture period. Specific growth rates of 0.13 days(-1) by fresh weight and 0.15 days(-1) by dry weight were measured. Elicitation with methyl jasmonate (MJ) significantly decreased both cell cycle progression and biomass accumulation, as the specific growth rate decreased to 0.027 days(-1) by fresh and dry weight. Despite the decrease in biomass accumulation for MJ elicited cultures, sucrose utilization was not significantly different from control cultures. MJ elicitation also increased the accumulation of paclitaxel and other taxanes. The accumulation of upstream taxanes (baccatin III and 10-deactylbaccatin III) increased during exponential growth, reached a maximum around day 12, and then declined throughout the stationary phase. The paclitaxel concentration increased during both exponential growth and stationary phase, reaching a maximum around days 20-25. Throughout the culture period, greater than 70% of the cells were in G(0)/G(1) phase of the cell cycle. Studies using bromodeoxyuridine (BrdU) incorporation showed that approximately 65% of the Taxus cells are noncycling, even during exponential growth. Although the role of these cells is currently unknown, the presence of a large, noncycling subpopulation can have a significant impact on the utilization of plant cell culture technology for the large-scale production of paclitaxel. These results demonstrate that there is a high degree of metabolic heterogeneity in Taxus cuspidata suspension cultures. Understanding this heterogeneity is important for the optimization of plant cell cultures, particularly the reduction of production variability. PMID- 15812806 TI - Renal frame count: a quantitative angiographic assessment of renal perfusion. AB - Renal perfusion may be measured by a variety of noninvasive methods; however, there is no objective angiographic method to assess renal perfusion. We measured the renal frame count (RFC) in 26 patients (50 kidneys) with normal renal function and normal renal angiograms and 9 patients (15 kidneys) with renal artery fibromuscular dysplasia (FMD) and normal renal function. The mean age of the patients with normal renal arteries was 61.5 +/- 9.5 (range, 47-82 years) and the mean age of patients with FMD was 72.5 +/- 9.2 (range, 54-86 years; P = 0.005). There was no correlation between the age and RFC in both the normal renal artery group and the FMD group. The mean RFC for the normal renal arteries was 20.4 +/- 3 (95% CI = 19.5-21.2), which was significantly lower than the FMD group's mean RFC of 26.9 +/- 9.9 (95% CI = 21.4-32.4; P = 0.0001). RFC is an objective angiographic measure to quantify renal perfusion. Compared to normal renal arteries, those with FMD had significantly increased RFC consistent with decreased perfusion. PMID- 15812807 TI - Percutaneous transluminal septal myocardial ablation for hypertrophic obstructive cardiomyopathy: how much alcohol should be injected? PMID- 15812808 TI - Peripheral vascular disease: carotid and vertebral brachytherapy for in-stent restenosis. AB - For years, intra-arterial brachytherapy has been a recognized method for treating recurrent stenosis after percutaneous transluminal angioplasty and stent placement. However, its use in arteries supplying the brain has not been described to date. We report a case treated with intra-arterial brachytherapy for high-grade recurrent stenoses of the high cervical internal carotid artery and the proximal vertebral artery. At 2-year follow-up, the outcome was successful. PMID- 15812809 TI - Congenital superior vena cava obstruction causing anasarca and respiratory failure in a newborn: successful transcatheter therapy. AB - Superior vena cava (SVC) obstruction is a rare entity in the pediatric population. It usually presents in association with either previous cardiac surgery or external compression from a neoplasm. We present the case of an infant born with congenital SVC obstruction and significant bilateral chylothorax and anasarca necessitating mechanical ventilation. Successful placement of an intravascular stent led to resolution of the chylothoraces with rapid clinical improvement. PMID- 15812810 TI - Whom do you trust? Misguided faith in the catheter- or Doppler-derived aortic valve gradient. PMID- 15812811 TI - Impact of different definitions on the interpretation of coronary remodeling determined by intravascular ultrasound. AB - The objective of this study was to compare the categorizations and determinants related to remodeling by the three definitions commonly used. Several morphological and intravascular ultrasound (IVUS) studies have demonstrated the fundamental importance of arterial remodeling in atherosclerosis. However, lack of consensus on how to define remodeling has led to conflicting analyses of factors that influence this process. Analysis of pre-interventional IVUS images of 514 lesions in native coronary arteries was performed. Arterial remodeling was defined as outward by definition 1, when [cross-sectional area (CSA) of the external elastic membrane (EEM) at the lesion site (EEM(lesion))]/[EEM CSA either at the proximal (EEM(prox ref)) or distal (EEM(distal ref)) reference site with the least amount of plaque] was > 1.05, intermediate when this ratio was between 0.95 and 1.05, and inward when < 0.95. Remodeling was defined as outward by definition 2 when EEM(lesion) > both EEM(prox ref) and EEM(distal ref), inward when EEM(lesion) < both EEM(prox ref) and EEM(distal ref), and intermediate when EEM(lesion) was intermediate between EEM(prox ref) and EEM(distal ref). By definition 3, vessel remodeling was defined as outward when EEM(lesion) > (EEM(prox ref) + EEM(distal ref))/2 and intermediate/inward when EEM(lesion) < or = (EEM(prox ref) + EEM(distal ref))/2. The frequency of outward remodeling was significantly higher by definitions 1 and 3 than by definition 2, whereas a higher frequency of inward remodeling was observed in definition 1, resulting in significantly different remodeling distributions between the three definitions (P < 0.0001). By multivariate logistic analysis, the only clinical determinants related to outward remodeling was younger age, and only by definition 3. IVUS determinants varied significantly between the three definitions. The only consistent determinants among the three definitions were smaller lumen CSA at the reference site and larger plaque + media CSA at the lesion site. This study demonstrates the significant impact of different remodeling definitions on the incidence and determinants of remodeling patterns. The marked variability in categorization of remodeling underscores the importance of developing a standard methodology. PMID- 15812812 TI - SOX2 anophthalmia syndrome. AB - Heterozygous, de novo, loss-of-function mutations in SOX2 have been shown to cause bilateral anophthalmia. Here we provide a detailed description of the clinical features associated with SOX2 mutations in the five individuals with reported mutations and four newly identified cases (including the first reported SOX2 missense mutation). The SOX2-associated ocular malformations are variable in type, but most often bilateral and severe. Of the nine patients, six had bilateral anophthalmia and two had anophthalmia with contralateral microphthalmia with sclerocornea. The remaining case had anophthalmia with contralateral microphthalmia, posterior cortical cataract and a dysplastic optic disc, and was the only patient to have measurable visual acuity. The relatively consistent extraocular phenotype observed includes: learning disability, seizures, brain malformation, specific motor abnormalities, male genital tract malformations, mild facial dysmorphism, and postnatal growth failure. Identifying SOX2 mutations from large cohorts of patients with structural eye defects has delineated a new, clinically-recognizable, multisystem disorder and has provided important insight into the developmental pathways critical for morphogenesis of the eye, brain, and male genital tract. PMID- 15812813 TI - Overestimation of aortic valve gradient measured by Doppler echocardiography in patients with aortic stenosis. AB - We describe a series of patients with suspected aortic stenosis who were noted to have a significant aortic valve gradient by Doppler echocardiography but only a minimal gradient during cardiac catheterization. Although the pressure recovery phenomenon partially explained this discrepancy, a difference still persisted, particularly in peak gradients. The overestimation occurred at much lower cardiac catheterization gradients than previously reported. Since many of our patients were noted to be hypertensive, this discrepancy might be more apparent in this patient population. PMID- 15812814 TI - Astrocyte-associated axonal damage in pre-onset stages of experimental autoimmune encephalomyelitis. AB - Recent studies of axon-glia and glia-glia communication have emphasized interactivity and interdependence between central nervous system (CNS) components. Concurrently, data from imaging, biochemical, and morphological studies have changed the view of multiple sclerosis (MS) from a neuroinflammatory condition with primary demyelination to one in which cumulative axonal damage drives progression. We therefore studied axonal damage in the context of inflammation and glial responses, from the pre-clinical to onset stage of murine experimental autoimmune encephalomyelitis (EAE), an established MS model. We report three major findings: (1) the first evidence of axonal injury before significant T-cell entry into the parenchyma, (3) coincidence of the earliest manifestation of axonal damage and astrocytic responses, and (3) an association between accumulation of axonal and astrocytic changes and specific forms of MS. These data demonstrate the relationship between the initiation of axonal injury and early inflammation. Significantly, we show that, in common with a growing number of neurodegenerative conditions, the pathology of murine EAE is characterized by early active contribution from astrocytes. This marks a change in the understanding of the role of astrocytes in MS pathogenesis and has important implications for the development of neuroprotective strategies. PMID- 15812815 TI - Schwann cells express erythropoietin receptor and represent a major target for Epo in peripheral nerve injury. AB - Erythropoietin (Epo) expresses potent neuroprotective activity in the peripheral nervous system; however, the underlying mechanism remains incompletely understood. In this study, we demonstrate that Epo is upregulated in sciatic nerve after chronic constriction injury (CCI) and crush injury in rats, largely due to local Schwann cell production. In uninjured and injured nerves, Schwann cells also express Epo receptor (EpoR), and its expression is increased during Wallerian degeneration. CCI increased the number of Schwann cells at the injury site and the number was further increased by exogenously administered recombinant human Epo (rhEpo). To explore the activity of Epo in Schwann cells, primary cultures were established. These cells expressed cell-surface Epo receptors, with masses of 71 and 62 kDa, as determined by surface protein biotinylation and affinity precipitation. The 71-kDa species was rapidly but transiently tyrosine phosphorylated in response to rhEpo. ERK/MAP kinase was also activated in rhEpo treated Schwann cells; this response was blocked by pharmacologic antagonism of JAK-2. RhEpo promoted Schwann cell proliferation, as determined by BrdU incorporation. Cell proliferation was ERK/MAP kinase-dependent. These results support a model in which Schwann cells are a major target for Epo in injured peripheral nerves, perhaps within the context of an autocrine signaling pathway. EpoR-induced cell signaling and Schwann cell proliferation may protect injured peripheral nerves and promote regeneration. PMID- 15812816 TI - Theodor Lipps and the shift from "sympathy" to "empathy". AB - In the course of extensive philosophical debates on aesthetics in nineteenth century Germany, Robert Vischer introduced the concept of Einfuhlung in relation to art. Theodor Lipps subsequently extended it from art to visual illusions and interpersonal understanding. While Lipps had regarded Einfuhlung as basically similar to the old notion of sympathy, Edward Titchener in America believed it had a different meaning. Hence, he coined the term empathy as its translation. This term came to be increasingly widely adopted, first in psychology and then more generally. But the lack of agreement about the supposed difference between these concepts suggests that Lipps had probably been right. PMID- 15812819 TI - Operationism in psychology: what the debate is about, what the debate should be about. AB - I offer an analysis of operationism in psychology, which is rooted in an historical study of the investigative practices of two of its early proponents (S. S. Stevens and E. C. Tolman). According to this analysis, early psychological operationists emphasized the importance of experimental operations and called for scientists to specify what kinds of operations were to count as empirical indicators for the referents of their concepts. While such specifications were referred to as "definitions," I show that such definitions were not taken to constitute a priori knowledge or be analytically true. Rather, they served the pragmatic function of enabling scientists to do research on a purported phenomenon. I argue that historical and philosophical discussions of problems with operationism have conflated it, both conceptually and historically, with positivism, and I raise the question of what are the "real" issues behind the debate about operationism. PMID- 15812817 TI - Neuregulin-1 enhances survival of human astrocytic glioma cells. AB - Malignant astrocytic gliomas, referred to as astrocytomas, represent the most commonly diagnosed adult primary brain tumor. These tumors are characterized by unrelenting growth that is often resistant to chemotherapy and radiation therapy. Tumor expansion into the healthy surrounding brain tissue produces severe and often fatal consequences. In this study, we examine the potential for the neuregulin-1/erbB receptor signaling cascade to contribute to this process by modulating glioma cell growth. Using antibodies specific for the erbB receptors, we demonstrate the expression patterns for the erbB2, erbB3, and erbB4 receptors in human glioma biopsy samples. We then verify receptor expression in a panel of human glioma cell lines. Next, we investigate the status of the erbB2 and erbB3 receptors in the human glioma cell lines and find that they are constitutively tyrosine-phosphorylated and heterodimerized. Subsequently, we demonstrate that theses same cell lines express membrane bound and released forms of neuregulins, the erbB receptor ligands, suggesting a possible autocrine or paracrine signaling network. Furthermore, we show that exogenous activation of erbB2 and erbB3 receptors in U251 glioma cells by recombinant Nrg-1beta results in enhanced glioma cell growth under conditions of serum-deprivation. This enhancement is due to an increase in cell survival rather than an increase in cell proliferation and is dependent on the activation of erbB2 and phosphatidylinositol-3 kinase (PI3K). Moreover, Nrg-1beta activates an inhibitor of apoptosis, Akt, implying a possible role for this kinase in mediating Nrg-1beta effects in gliomas. This data suggests that glioma cells may use autocrine or paracrine neuregulin-1/erbB receptor signaling to enhance cell survival under conditions where growth would otherwise be limited. PMID- 15812820 TI - The early origins and development of the scatterplot. AB - Of all the graphic forms used today, the scatterplot is arguably the most versatile, polymorphic, and generally useful invention in the history of statistical graphics. Its use by Galton led to the discovery of correlation and regression, and ultimately to much of present multivariate statistics. So, it is perhaps surprising that there is no one widely credited with the invention of this idea. Even more surprising is that there are few contenders for this title, and this question seems not to have been raised before. This article traces some of the developments in the history of this graphical method, the origin of the term scatterplot, the role it has played in the history of science, and some of its modern descendants. We suggest that the origin of this method can be traced to its unique advantage: the possibility to discover regularity in empirical data by smoothing and other graphic annotations to enhance visual perception. PMID- 15812822 TI - Imatinib mesylate lacks activity in small cell lung carcinoma expressing c-kit protein: a phase II clinical trial. AB - BACKGROUND: Imatinib inhibits the c-kit tyrosine kinase, which, accounts for its activity in gastrointestinal stromal tumors. The presence of c-kit protein expression in small cell lung carcinoma (SCLC) tumor specimens, as well as in vitro data supporting the role of c-kit in autocrine and paracrine growth stimulation specifically in SCLC, provided a rationale for studying imatinib in this disease. The authors conducted a Phase II single-institution study of imatinib in patients with recurrent SCLC whose tumor specimens expressed c-kit protein. METHODS: Patients with progressive SCLC after one or two previous chemotherapy regimens consented to have their tumor specimens screened by immunoperoxidase stain (CD117, Dako Corporation, Carpinteria, CA) for c-kit protein expression. If present, individuals were then eligible for treatment with an imatinib dose of 400 mg orally twice daily (total, 800 mg per day). RESULTS: The presence of c-kit protein was assessable in 36 of 39 (92%) tumor samples. Twenty-eight (78%) tumor samples had immunohistochemical staining for c-kit protein. Twelve patients were enrolled in the treatment portion of the current study. No responses were observed, and all patients had disease progression by Week 4. Edema, fatigue, nausea, and electrolyte abnormalities were the primary toxicities. CONCLUSIONS: Imatinib did not have antitumor activity against SCLC, even with c-kit protein present in tumor specimens. The dismal prognosis for these patients with progressive SCLC emphasized the urgent need for continued studies of new therapies in this population. PMID- 15812823 TI - Survivors of childhood cancer and their guardians. AB - BACKGROUND: Survivors of childhood cancer are at increased risk for osteoporosis, cardiovascular disease, and second malignancies-conditions for which modifiable risk factors are recognized and lifestyle interventions have shown benefit. Although some data regarding health behaviors of this population exist, receptivity to health promotion is largely unknown. METHODS: A survey was mailed to 380 survivors (age range, 11-33 years) of childhood leukemia, lymphoma, or central nervous system carcinomas (and guardians of survivors < 18 years old) to elicit data on exercise, dietary intake of calcium, fat, and fruits and vegetables, smoking status, readiness to pursue lifestyle change, quality of life, and interest in various health interventions. RESULTS: Responses from 209 survivors (a 55% response rate) suggested that most did not meet guidelines for fruit and vegetable consumption (79%), calcium intake (68%), or exercise (52%), 42% were overweight/obese, and 84% consumed > 30% of calories from fat. Older (> 18 years) compared with younger (< 18 years) survivors were more likely to smoke (17% vs. 1%), to be obese (21.6% vs. 14.6%), and to have suboptimal calcium intakes (75.6% vs. 57.6%). No differences in lifestyle behaviors were observed between cancer groups. Compared with interventions aimed at weight control, improving self-esteem, or smoking cessation, the highest levels of interest were found consistently for interventions aimed at getting in shape and eating healthy. Survivors preferred mailed interventions to those delivered in-person, by telephone counselors, or via computers. CONCLUSIONS: Survivors of childhood cancer practiced several suboptimal health behaviors. Health promotion interventions aimed at areas of interest and delivered through acceptable channels have the potential to improve long-term health and function of this vulnerable population. PMID- 15812825 TI - The prognostic significance of the percentage of positive/dissected axillary lymph nodes in breast cancer recurrence and survival in patients with one to three positive axillary lymph nodes. AB - BACKGROUND: Adjuvant therapy for women with T1-T2 breast carcinoma and 1-3 positive lymph nodes is controversial due to discrepancies in reported baseline locoregional recurrence (LRR) risks. This inconsistency has been attributed to variations in lymph node staging techniques, which have yielded different numbers of dissected lymph nodes. The current study evaluated the prognostic impact of the percentage of positive/dissected lymph nodes on recurrence and survival in women with one to three positive lymph nodes. METHODS: The study cohort was comprised of 542 women with pathologic T1-T2 breast carcinoma who had 1-3 positive lymph nodes and who had undergone mastectomy and received adjuvant systemic therapy without radiotherapy. Ten-year Kaplan-Meier (KM) LRR, distant recurrence (DR), and overall survival (OS) rates stratified by the number of positive lymph nodes, the number of dissected lymph nodes, and the percentage of positive lymph nodes were examined using different cut-off levels. Multivariate analysis was performed to evaluate the prognostic significance of the percentage of positive lymph nodes in disease recurrence and survival. RESULTS: The median follow-up was 7.5 years. LRR, DR, and OS rates correlated significantly with the number of positive lymph nodes and the percentage of positive lymph nodes, but not with the number of dissected lymph nodes. The cut-off level at which the most significant difference in LRR was observed was 25% positive lymph nodes (the 10 year KM LRR rates were 13.9% and 36.7% in women with < or = 25% and > 25% positive lymph nodes, respectively; P < 0.0001). Higher DR rates and lower OS rates were observed among patients who had > 25% positive lymph nodes compared with patients who had < or = 25% positive lymph nodes (DR: 53.0% vs. 30.3%, respectively; P < 0.0001; OS: 43.4% vs. 62.6%, respectively; P < 0.0001). In the multivariate analysis, the percentage of positive lymph nodes and the histologic grade were significant, independent factors associated with LRR, DR, and OS. CONCLUSIONS: The presence of > 25% positive lymph nodes was an adverse prognostic factor in patients with 1-3 positive nodes and may be used to identify patients at high risks of postmastectomy locoregional and distant recurrence who may benefit with adjuvant radiotherapy and more aggressive systemic therapy regimens. PMID- 15812824 TI - Hepatomegaly in transgenic mice expressing the homeobox gene Cux-1. AB - Cux-1 is a member of a family of homeobox genes structurally related to Drosophila Cut. Mammalian Cut proteins function as transcriptional repressors of genes specifying terminal differentiation in multiple cell lineages. In addition, mammalian Cut proteins serve as cell-cycle-dependent transcriptional factors in proliferating cells, where they function to repress expression of the cyclin kinase inhibitors p21 and p27. Previously we showed that transgenic mice expressing Cux-1 under control of the CMV immediate early gene promoter develop multiorgan hyperplasia. Here we show that mice constitutively expressing Cux-1 exhibit hepatomegaly correlating with an increase in cell proliferation. In addition, the increase in Cux-1 expression in transgenic livers was associated with a decrease in p21, but not p27, expression. Within transgenic livers, Cux-1 was ectopically expressed in a population of small cells, but not in mature hepatocytes, and many of these small cells expressed markers of proliferation. Transgenic livers showed an increase in alpha-smooth muscle actin, indicating activation of hepatic stellate cells, and an increase in cells expressing chromogranin-A, a marker for hepatocyte precursor cells. Morphological analysis of transgenic livers revealed inflammation, hepatocyte swelling, mixed cell foci, and biliary cell hyperplasia. These results suggest that increased expression of Cux-1 may play a role in the activation of hepatic stem cells, possibly through the repression of the cyclin kinase inhibitor p21. PMID- 15812827 TI - Differential release of cardiac enzymes after percutaneous coronary intervention. AB - We hypothesized that using calcium channel blockers (CCBs) that dilate microvasculature during percutaneous coronary intervention (PCI) would result in lower postprocedural creatine phosphokinase (CPK). PCI can be complicated by elevated CPK that has been associated with impaired microvascular perfusion. Nitroglycerin (NTG), the conventional PCI vasodilator, dilates epicardial arteries but does not affect the microvasculature. We hypothesized that using CCBs that dilate the microvasculature would result in lower postprocedural CPK values. Patients (n = 816) without evidence of acute myonecrosis undergoing PCI were divided into two groups based on whether they received intracoronary NTG or CCB during PCI. Postprocedural CPK values were compared using a repeated-measures ANOVA and a random coefficient model. By repeated-measures analysis, the NTG group had CPK values of 88%, 83%, and 89% of the CCB group's CPK values at < 8, 8 14, and > 14 hr after PCI (P = 0.0080, 0.0002, and 0.0244), respectively. In a random coefficient model, the NTG group had CPK values 84%, 84%, and 89% of the CCB group's mean CPK values at 6, 12, and 18 hr after PCI (P = 0.0003, 0.0006, and 0.0403), respectively. Peak CPK values occurred earlier with CCB, although the maximal CPK was similar in both groups. Intracoronary CCB use is associated with an accelerated release of CPK after PCI compared with NTG. This is consistent with more efficient relief of microvascular obstruction with CCB. It suggests that myonecrosis may originate with vascular trauma at the time of PCI and its enzymatic expression is modifiable with different vasodilators. PMID- 15812828 TI - CD13 (aminopeptidase N) can associate with tumor-associated antigen L6 and enhance the motility of human lung cancer cells. AB - Cancer metastasis is a multiple-step process that involves the regulated interaction of diverse cellular proteins. We recently reported that the expression of tumor-associated antigen L6 (TAL6) promoted the invasiveness of lung cancer cells and was inversely correlated with disease-free survival of squamous lung carcinoma patients. We now report that CD13 (aminopeptidase N) can associate with TAL6 and can enhance cancer cell migration. CD13 was shown by coimmunoprecipitation to associate in vitro with TAL6 on several cancer cell lines and to associate in vivo by antibody-mediated copatching immunofluorescence. CD13 was selectively expressed on highly invasive CL1-5 lung cancer cells as compared to poorly invasive CL1-0 lung cancer cells. The role of CD13 aminopeptidase activity in regulating cell motility was investigated with chemical inhibitors, specific antibodies and a catalytically inactive CD13 protein. Inhibition of CD13 aminopeptidase activity by nontoxic concentrations of leuhistin modestly decreased the migration of CL1-5 cells. In contrast, binding of CD13 by specific antibodies significantly reduced both the migration and the invasion of CL1-5 cells. Poorly invasive CL1-0 cells that stably expressed CD13 displayed significantly (p < or = 0.0005) enhanced cell migration (300% of control). Expression of an enzymatically inactive CD13 mutant on CL1-0 cells also significantly (p < or = 0.0005) enhanced cell migration (200% of control). Our results show that TAL6 and CD13 can form a complex on lung cancer cells, that these molecules can modulate cell migration and invasion and that the influence of CD13 on cell motility did not strictly depend on its aminopeptidase activity. PMID- 15812830 TI - Structural characterization of human elastin derived peptides containing the GXXP sequence. AB - The degradation of elastin, the insoluble biopolymer of tropoelastin, can lead to the production of small peptides. These elastin-derived peptides (EDPs) are playing a key role in cellular behavior within the extracellular matrix, showing a great variety of biological effects such as chemotaxis, stimulation of cell proliferation, ion flux modifications, vasorelaxation, and inflammatory enzymes secretion. It has also been demonstrated recently that EDPs containing the GXXPG motif could induce pro-MMP1 and pro-MMP3 upregulation. Elastolysis could then cause collagen degradation and play an important role in the aging process. Many experimental studies have been devoted to EDPs, but their structure/activity relationships are not well elucidated yet. However, the assumption that their active conformation is a type VIII beta-turn on GXXP was highly suggested on the basis of predictive statistical calculations. Investigation of the EDPs three dimensional (3D) structure would provide useful information for drug-design strategies to propose specific inhibitors. The work presented here reports theoretical results obtained from molecular dynamics simulations performed over 128 human EDPs containing the GXXP motif. We show that all the peptides, for which the central residues are not glycines, adopt a canonical (or very close to) type VIII beta-turn structure on the GXXP sequence. Amino acids surrounding this motif are also important for the structural behavior. Any residue located before the GXXP motif (XGXXP) increases the beta-turn stabilization, whereas the residue located after GXXP (GXXPX) has no significant structural effect. Moreover, we show their biological activity can be correlated with their ability to exhibit a type VIII beta-turn conformation. PMID- 15812831 TI - Blockheads, roundheads, pointy heads: intellectual disability and the brain before modern medicine. AB - Recent work on the conceptual history of intellectual disability has pointed to a discontinuity in the seventeenth century, identifying the concept as essentially modern in a more radical sense than mental illness or physical disability. However, Galenist accounts of intellectual impairment were clearly connected (via anatomy) to neurology, which could be taken as prima facie evidence that Galenism shares with modern medicine one of its basic explanatory approaches to intellectual disability. Close textual examination does not bear out this counter claim, at least as far as the conceptual apparatus itself is concerned. However, it does reveal a degree of continuity in the medical mind-set as discourses of monstrosity were transposed from the domain of anatomy to that of post-Cartesian psychology. PMID- 15812832 TI - Mucinous carcinomas of the colon and rectum show higher rates of microsatellite instability and lower rates of chromosomal instability: a study matched for T classification and tumor location. AB - BACKGROUND: The clinicopathologic significance of mucinous carcinomas (Muc) of the colon and rectum has been widely discussed, but there have been few studies on Muc regarding genetic and epigenetic alterations. The current study analyzed genetic and epigenetic alterations of Muc to clarify their differences from well differentiated adenocarcinomas (WD). METHODS: Thirty-nine cases of Muc and 39 cases of WD were investigated. Cases of WD were matched with cases of Muc for T classification and tumor location. Microsatellite instability (MSI) status and loss of heterozygosity (LOH) of four loci (2p, 5q, 17p, 18q) were evaluated. The methylation status of the hMLH1 promoter region in Muc was also examined. RESULTS: "MSI tumors" were defined as those that showed MSI-high, and "chromosomal instability (CIN) tumors" were defined as those that showed LOH but not MSI-high. MSI tumors were significantly more frequent in Muc (30.8%) than in WD (5.1%). CIN tumors were significantly less frequent in Muc (53.8%) than in WD (87.2%). In Muc, MSI tumors were significantly more frequent in the proximal colon (55.6%) than in the distal colon (9.5%). Also, methylation of the hMLH1 promoter region in Muc was significantly more frequent in MSI tumors (83.3%) than in CIN tumors (27.8%) (P = 0.0077). CONCLUSIONS: When matched for T classification and tumor location, Muc shows higher rates of MSI and lower rates of CIN than WD.. Muc shows different characteristics according to tumor location, and methylation of the hMLH1 promoter region strongly correlates with Muc tumors showing MSI. PMID- 15812833 TI - A phase II trial of R115777, an oral farnesyl transferase inhibitor, in patients with advanced urothelial tract transitional cell carcinoma. AB - BACKGROUND: R115777 is a potent farnesyl transferase inhibitor and has significant antitumor effects in vitro and in vivo. METHODS: The objective of the current study was to determine the objective response proportion in patients with metastatic transitional cell carcinoma (TCC) of the urothelial tract who received treatment with R115777 at a dose of 300 mg orally given twice daily for 21 days followed by 7 days of rest for every 4-week cycle. Thirty-four patients with TCC were enrolled in this Phase II study. Patients were allowed to have received a maximum of one prior systemic chemotherapy regimen, not including chemoradiation or neoadjuvant chemotherapy. All patients were required to have an Eastern Cooperative Oncology Group performance status of 0-2 and adequate bone marrow, hepatic, and kidney function. RESULTS: Twice daily administration of oral R115777 was tolerated well. R115777 was absorbed rapidly after oral administration. Grade 3-4 neutropenia (according to the National Cancer Institute Common Toxicity Criteria [version 2.0]) was observed in 5 patients (15%). Grade 3-4 nonhematologic toxicity was rare, consisting of rash and diarrhea in 1 patient each. Two patients (6%) without prior chemotherapy demonstrated partial responses. Thirteen patients (38%) achieved disease stabilization according to World Health Organization criteria that lasted a median of 4 months. No complete responses were observed. CONCLUSIONS: The objective response rate of R115777 was not sufficient to warrant future investigation in TCC as a single agent. Preliminary evidence of the activity of R115777 in 2 chemotherapy-naive patients may warrant further investigation in combination with first-line chemotherapy. PMID- 15812834 TI - Hypofractionated three-dimensional conformal radiation therapy for primary liver carcinoma. AB - BACKGROUND: The purpose of the current study was to evaluate the tolerance and efficacy of hypofractionated three-dimensional conformal radiotherapy (3DCRT) with or without transarterial chemoembolization (TACE) for technically unresectable or medically inoperable primary liver carcinoma (PLC). METHODS: Between April 1999 and August 2003, 128 patients with a clinical diagnosis of PLC received hypofractionated 3DCRT at Cancer Hospital, Guangxi Medical University. Both hypofractionated 3DCRT and TACE were used to treat 48 of these 128 patients. Liver cirrhosis of Child-Pugh Grade A was found in 108 patients, and Grade B was found in 20 patients. The mean gross tumor volume (GTV) was 459 +/- 430 cm3. A mean total irradiation dose of 53.6 +/- 6.6 Gy was delivered at an average fraction of 4.88 +/- 0.47 Gy, 3 times a week using 8-MV photons. RESULTS: The median follow-up time after 3DCRT was 12 months (range, 2-56 mos.). The immediate response rate was 55%. The overall survival rates at 1, 2, and 3 years were 65%, 43%, and 33%, respectively, with a median survival of 20 months (range, 7-31 mos.). Radiation Therapy Oncology Group (RTOG) Grade 2 acute gastrointestinal complications developed in 8 patients, whereas 4 patients developed Grade 3 late gastrointestinal complications. Radiation-induced liver disease (RILD) developed in 19 (15%) patients, of which 12 had Child-Pugh Grade B liver cirrhosis, and 7 had Grade A. GTV and associated liver cirrhosis were identified by Cox regression analysis as independent predictors for survival (P = 0.044 and 0.015). CONCLUSIONS: Hypofractionated 3DCRT is effective in carefully selected patients with PLC. Gastrointestinal complications and RILD were the most distinct complications. PMID- 15812835 TI - System zones in capillary zone electrophoresis: moving boundaries caused by freely migrating hydroxide ions. AB - We present theoretical and experimental data indicating that anionic system zones (SZs), due to free migrating hydroxide anions, can be expected in background electrolytes (BGEs) with a low buffer capacity. In the system containing completely unbuffered BGEs the hydroxide ions derived from the sample start to migrate freely through the capillary tube with the mobility of single hydroxide ions and cause stepwise disturbances in the baseline of the detector trace. Remarkably, this type of SZs do not appear to contribute significantly to the electromigration dispersion (EMD) of the zones of the analytes. PMID- 15812836 TI - Polymethacrylate-type monoliths functionalized with chiral amino phosphonic acid derived strong cation exchange moieties for enantioselective nonaqueous capillary electrochromatography and investigation of the chemical composition of the monolithic polymer. AB - In situ prepared monolithic poly(glycidyl methacrylate-co-ethylene dimethacrylate) (poly(GMA-co-EDMA)) capillary columns were activated to reactive thiol-monoliths and subsequently functionalized with (S)-N-(4-allyloxy-3,5 dichlorobenzoyl)-2-amino-3,3-dimethylbutanephosphonic acid as chiral selector by radical addition to afford enantioselective strong cation exchanger (SCX) capillary columns (100 microm inner diameter (ID)). These monolithic capillaries were devised for the enantioseparation of chiral bases by nonaqueous and aqueous capillary electrochromatography (CEC) and the results obtained for mefloquine and its tert-butylcarbamate as test compounds were compared to those obtained with particulate silica-based analogs (packed columns). Despite abolishment of nonspecific ionic interactions between the cationic solutes and residual silanols that may diminish separation factors of the silica-based chiral SCX particles, the poly(GMA-co-EDMA)-supported SCX monolith did not, as expected, show better enantioselectivities, which was assumed to be due to detrimental nonspecific interactions between the analytes and the lipophilic polymer backbone. In order to minimize these unfavorable contributions, less lipophilic monoliths were developed by copolymerization of different amounts of the hydrophilic monomer 2 hydroxyethyl methacrylate (HEMA) with GMA and EDMA, leading to GMA-co-HEMA-co EDMA-terpolymeric monoliths. By this increase of the hydrophilicity of the monolithic support the enantioselectivity of the resultant SCX stationary phase could be enhanced and reached values comparable to the packed silica-based enantioselective SCX capillaries. Additionally, the mobile phase composition and other variables were examined and it could be shown that the separation factors are considerably affected by diverse parameters such as acetonitrile-methanol ratio and type and concentration of the counterion. Mefloquine enantiomers could be separated with alpha-values up to 1.56 and a maximum plate count of ca. 60,000 m(-1) could be achieved. PMID- 15812837 TI - Thiol redox status evaluation in red blood cells by capillary electrophoresis laser induced fluorescence detection. AB - Thiols and in particular glutathione (GSH) play a central role in human metabolism, including the detoxification of xenobiotics, cell homeostasis, radioprotection, and antioxidant defence. Here, a new method is provided for the measurement of reduced and total forms of thiols in red blood cells. In order to minimize oxidation of reduced thiols, a water erythrocyte lysis (15 min at 4 degrees C) was performed followed by a protein precipitation step with acetonitrile. The supernatant was rapidly derivatized with 5 iodoacetoamidefluorescein that trapped thiol groups, thus minimizing auto oxidation. Derivatized samples were separated in a 57 cm x 75 microm ID capillary by using 5 mmol/L sodium phosphate, 4 mmol/L boric acid as electrolyte solution with 75 mmol/L N-methyl-D-glucamine at pH 11.0. Under these conditions, cysteinylglycine (CysGly), cysteine (Cys), glutathione, and gamma glutamylcysteine (GluCys) were baseline-resolved in approximately 4 min. Precision tests showed a good repeatability of our method both for migration times (coefficient of variation CV < 0.8%) and areas (CV < 3.3%). Furthermore, a good reproducibility of intrassay and interassay tests was obtained (CV < 5% and CV < 8%, respectively). The method was employed to investigate the effect of acidic precipitation on intracellular thiol concentration. Our data suggest that sample acidification causes a modification of the measured redox thiol status due to the development of a pro-oxidant environment; moreover, the thiol redox status of red blood cells was evaluated in 22 healthy volunteers. PMID- 15812838 TI - A polymeric master replication technology for mass fabrication of poly(dimethylsiloxane) microfluidic devices. AB - A protocol of producing multiple polymeric masters from an original glass master mold has been developed, which enables the production of multiple poly(dimethylsiloxane) (PDMS)-based microfluidic devices in a low-cost and efficient manner. Standard wet-etching techniques were used to fabricate an original glass master with negative features, from which more than 50 polymethylmethacrylate (PMMA) positive replica masters were rapidly created using the thermal printing technique. The time to replicate each PMMA master was as short as 20 min. The PMMA replica masters have excellent structural features and could be used to cast PDMS devices for many times. An integration geometry designed for laser-induced fluorescence (LIF) detection, which contains normal deep microfluidic channels and a much deeper optical fiber channel, was successfully transferred into PDMS devices. The positive relief on seven PMMA replica masters is replicated with regard to the negative original glass master, with a depth average variation of 0.89% for 26-microm deep microfluidic channels and 1.16% for the 90 mum deep fiber channel. The imprinted positive relief in PMMA from master-to-master is reproducible with relative standard deviations (RSDs) of 1.06% for the maximum width and 0.46% for depth in terms of the separation channel. The PDMS devices fabricated from the PMMA replica masters were characterized and applied to the separation of a fluorescein isothiocyanate (FITC)-labeled epinephrine sample. PMID- 15812839 TI - Urtica dioica agglutinin: separation, identification, and quantitation of individual isolectins by capillary electrophoresis and capillary electrophoresis mass spectrometry. AB - With benign prostatic hyperplasia (BPH) being a major health problem in ageing men, alternative therapeutic approaches (e.g., with phytopharmaceuticals) are of great interest. Based on pharmacological evidences, one of the most promising options in that respect are the lectins found in Urtica dioica (stinging nettle) roots. In this study the qualitative and quantitative analysis of individual isolectins in U. dioica extracts is described, which is the first report on using capillary electrophoresis (CE) for the analysis of lectins in plant material at all. By utilizing a 200 mM sodium acetate buffer (pH 3.75) a baseline separation and determination of four closely related isolectins was feasible within 20 min in the aqueous plant extracts. The individual compounds were identified based on reference compounds as well as data obtained from CE-mass spectrometry (MS) experiments. After modifying the optimized CE conditions to 100 mM ammonium formate buffer with pH 3.75 and a voltage of 15 kV, the isolectins were clearly assignable in positive electrospray ionization (ESI) mode. The quantitative results obtained by CE (the total lectin content varied from 0 to 0.42% in the samples) were accurate (recovery rates of spiked samples between 92.5 and 96.2%), precise (relative standard deviation < 5%) and in good agreement to those obtained by High-performance liquid chromatography (HPLC). As for peak resolution, assignable compounds and required separation time the newly developed CE method was clearly advantageous over the determination achieved by LC. PMID- 15812840 TI - The molecular end effect and its critical impact on the behavior of charged uncharged polymer conjugates during free-solution electrophoresis. AB - Recently two novel techniques using free-solution electrophoresis to separate charged-uncharged polymer conjugates have proven successful: end-labeled free solution electrophoresis (ELFSE) for DNA sequencing, and free-solution conjugate electrophoresis (FSCE) for molar mass profiling of uncharged polymers. The approach taken to analyze the experimental data was an extension of the theory of Long and co-workers (Long, D., Dobrynin, A. V., Rubinstein, M., Ajdari, A., J. Chem. Phys. 1998, 108, 1234-1244) for the electrophoresis of molecules with varying charge distributions. This theory also predicts that the ends of the polymers play a large role in determining the polymer's overall mobility; however, this aspect of the theory was neglected in previous work. Until now this "end effect" has, to the knowledge of the authors, not been recognized in experimental data. Through a careful investigation of the predicted end effect and a reanalysis of the experimental data, we demonstrate that indeed this effect critically impacts on the behavior of charged-uncharged polymer conjugates during electrophoresis. This work indicates that not only does the end effect need to be taken into account to avoid significant errors in data analysis, but also it provides novel system optimization approaches. PMID- 15812841 TI - A facile and sensitive chemiluminescence detection of amino acids in biological samples after capillary electrophoretic separation. AB - It was found that native amino acids enhanced the chemiluminescence (CL) reaction between luminol and BrO(-) in an alkaline aqueous solution. This has led to the development of a facile and highly sensitive CL detection scheme for the determination of amino acids in biological samples after capillary electrophoretic (CE) separation. The CE-CL conditions were optimized. An electrophoretic buffer of 2.5 x 10(-2) M sodium borate (pH 9.4) containing 1 x 10(-4) M luminol was used. The oxidizer solution of 8 x 10(-4) M NaBrO in 0.1 M sodium carbonate buffer solution (pH 12.5) was introduced post-column. Under the optimal conditions, the detection limits were 1.0 x 10(-7) M for glutamic acid (Glu) and 1.3 x 10(-7) M (S/N = 3) for aspartic acid (Asp). The relative standard deviations (RSDs) of peak area and migration time were in the ranges of 3.8-4.3% and 1.4-1.6%, respectively. The present method was applied to the determination of excitatory amino acids (i.e., Asp and Glu) in rat brain tissue and monkey plasma. The levels of these major excitatory amino acids in monkey plasma were quantified for the first time and found to be 1.17 +/- 0.17 x 10(-5) M (mean +/- SD, n = 6) for Glu and 1.64 +/- 0.19 x 10(-6) M for Asp, which were comparable with the levels in human plasma. PMID- 15812842 TI - Electrochemiluminescence quenching as an indirect method for detection of dopamine and epinephrine with capillary electrophoresis. AB - An electrochemiluminescence (ECL) inhibition method was developed as an indirect detection method for the determination of dopamine and epinephrine separated by capillary electrophoresis (CE). When the concentration of Ru(bpy)(3) (2+) was 50 muM diluted by 50 mM phosphate (pH 8.5) in the cell and 0.5 M tripropylamine (TPA) was added to the running buffer (10 mM phosphate, pH 9.0), an inhibition of ECL of the Ru(bpy)(3) (2+)/TPA system by the analytes was observed. Under the optimized conditions, the relative standard deviations of migration time and negative peak area were less than 1% and 3%, respectively, for 1 microM dopamine or 1 microM epinephrine (n = 10). Linear ranges of 0.1-10 microM for both analytes and the detection limits (signal-to-noise ratio S/N = 3) of 10 nM for dopamine and 30 nM for epinephrine were obtained. PMID- 15812843 TI - Microautosamplers for discrete sample injection and dispensation. AB - Microfluidic systems show considerable potential for use in the continuous reaction and analysis of biosamples for various applications, such as drug screening and chemical synthesis. Typically, microfluidic chips are externally connected with large-scale autosamplers to inject specific volumes of discrete samples in the continuous monitoring and analysis of multiple samples. This paper presents a novel microelectromechanical system (MEMS)-based autosampler capable of performing the discrete injection and dispensation of variable-volume samples. This microdevice can be integrated with other microfluidic devices to facilitate the continuous monitoring and analysis of multiple biosamples. By means of electroosmotic focusing and switching controlled by the direct application of electric sources on specific fluid reservoirs, a precise sample volume can be injected into the specified outlet port. Fluorescence dye images verify the performance of the developed device. An injection-and-washing scheme is developed to prevent cross-contamination during the continuous injection of different samples. This approach renders feasible the injection of several discrete samples using a single microchip. Compared to its large-scale counterparts, the developed microautosampler is compact in size, has low fabrication costs, is straightforward to control, and most importantly, is readily integrated with other microfluidic devices (e.g., microcapillary electrophoresis chips) to form a microfluidic system capable of the continuous monitoring and analysis of bioreactions. The proposed microautosampler could be promising towards realizing the micrototal analysis system (mu-TAS) concept. PMID- 15812844 TI - A note on non-parametric ANCOVA for covariate adjustment in randomized clinical trials. PMID- 15812845 TI - Multiplex reverse transcription-polymerase chain reaction combined with on-chip electrophoresis as a rapid screening tool for candidate gene sets. AB - Combining multiplex reverse transcription-polymerase chain reaction (mRT-PCR) with microfluidic amplicon analysis, we developed an assay for the rapid and reliable semiquantitative expression screening of 11 candidate genes for drug resistance in human malignant melanoma. The functionality of this approach was demonstrated by low interexperimental variations of amplicon quantities after endpoint analysis. When applied to RNA samples derived from drug-sensitive and resistant melanoma cell lines, mRT-PCR delivered results qualitatively concordant with data obtained from Northern blot and array analyses. The screening of additional melanoma cell lines resulted in distinct expression patterns for ten candidate genes. Our approach reveals a rapid and easy-to-handle alternative for candidate gene set evaluation from limited amounts of RNA. PMID- 15812846 TI - Capillary zone electrophoretic studies of interactions of some quaternary isoquinoline alkaloids with DNA constituents and DNA. AB - Capillary zone electrophoresis was applied for the investigation of interactions of some quaternary isoquinoline alkaloids, namely sanguinarine, chelerythrine, berberine, and jatrorrhizine, with DNA constituents and with DNA. None of these alkaloids attach covalently to nucleotides or to the whole DNA under physiological conditions. The interaction with DNA constituents is a noncovalent complexation based on weak intermolecular forces. Electrostatic attraction participates in the interaction but other types of intermolecular forces are involved as well. Cations were identified as the most probable interacting forms of the alkaloids. The interaction with compounds derived from purine was always stronger than those derived from pyrimidine. All alkaloids behaved analogously and similarly to ethidium bromide, the classic DNA intercalator. Stability constants K (in l.mol(-1)) for sanguinarine and chelerythrine in phosphate buffer of pH 7.4 (I(S) = 30 mM) ranged from tens to hundreds. PMID- 15812847 TI - Long- and short-haired Weimaraner dogs represent two populations of one breed. AB - Weimaraners represent an old breed of hunting dogs. Today, two coat types are commonly distinguished, the more common short-hair (SH) and the long-hair (LH) variety, the latter having arisen from the SH Weimaraners. In order to analyze genetic variation in the coat varieties, we genotyped nine single nucleotide polymorphisms (SNPs) of the ABCA4 gene locus as well as six highly variable microsatellites scattered over the canine genome in the SH and LH populations. Three out of nine SNPs showed two alleles, allelic frequencies at two of these polymorphic sites differed significantly between SH and LH Weimaraners. Haplotype diversities for the three informative SNPs revealed higher estimates for the SH (0.515) than for the LH variety (0.364). In addition, two of six microsatellite markers showed significant differences in allelic frequencies between SH and LH Weimaraners. Unexpectedly, genetic diversities for all but one microsatellite were greater in LH than in SH Weimaraners. Similarly, the mean intra-individual genetic distance based on microsatellite markers was more pronounced in the LH population (0.62 for SH vs. 0.65 for LH) suggesting again closer genetic relationships among SH than LH Weimaraners. Taken together, the results of SNP analysis can be interpreted as reflections of early breed development whereas microsatellites mirror rather recent breeding strategies in the Weimaraner populations. PMID- 15812848 TI - An integrated microfluidic device in polyester for electrophoretic analysis of amino acids. AB - A precolumn reaction chamber was integrated into a polyester microfluidic device with a miniaturized detection system. The reaction chamber was designed to be a zigzag channel, 70 microm in width, 8 mm in length, followed by a wider straight channel, 150 microm in width, 2 mm in length. The detection system is composed of an embedded light-emitting diode (LED), an integrated optical fiber, and a photomultiplier tube (PMT). A success in amino acid analysis using the integrated microchemical analysis device proved that the precolumn reaction chamber was compatible with the integrated detection system. Three kinds of amino acids, arginine, glycine, and phenylalanine, mixed and reacted with 7-fluoro-4 nitrobenzo-2-oxa-1,3-diazole (NBD-F) in the precolumn reaction chamber to produce fluorescent products, were separated by micellar eletrokinetic chromatography (MEKC) and detected by LED-excited fluorescence. The detection limits for arginine, glycine, and phenylalanine were 1, 1, and 0.5 mM, respectively, which can be improved by further optimizations of the reaction system and detection system. PMID- 15812849 TI - The zeta potential of cyclo-olefin polymer microchannels and its effects on insulative (electrodeless) dielectrophoresis particle trapping devices. AB - While cyclo-olefin polymer microchannels have the potential to improve both the optical detection sensitivity and the chemical resistance of polymer microanalytical systems, their surface properties are to date not thoroughly characterized. These surface properties dictate, among other things, electrokinetic effects when electric fields are present. Here, we report the measurement of the zeta potential of cyclo-olefin polymers (injection-molded and hot-embossed Zeonor 1060R and 1020R) microchannels as a function of pH, counter ion concentration, storage conditions, and chemical treatment in aqueous solutions both with and without EOF-suppressing additives. In contrast with previous reports, significant surface charge is measured, consistent with titration of charged sites with pK(a) = 4.8. Storage in air, acetonitrile, or aqueous solutions has relatively minor effects. While the source of the surface charge is unclear, chemical functionalization has shown that carboxylic acid groups are not present at the surface, consistent with the chemical structure of Zeonor. EOF-suppressing additives (hydroxypropylmethylcellulose) and conditioning in perchloric acid allow the surface charge to be suppressed. We demonstrate dielectrophoretic particle trapping devices in Zeonor 1060R substrates that show reduced trapping voltage thresholds as compared to previous implementations in glass. PMID- 15812850 TI - Direct fluorescent staining and analysis of proteins on microspheres using CBQCA. AB - BACKGROUND: General methods for accurate determination of microsphere surface protein loading are needed for applications from protein arrays to molecular assembly studies. Current methods include bulk absorption measurements of stained microspheres or use of known fluorescently tagged binding partners, which limit sensitivity and general applicability, respectively. METHODS: Microspheres bearing covalently coupled proteins were stained with 3-(4 carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) using different incubation times and dye concentrations to determine optimal staining conditions. The CBQCA fluorescence of microspheres (measured by flow cytometry) bearing known amounts of protein were used to generate standard curves of CBQCA fluorescence response versus the amount of microsphere surface protein. CBQCA was also used to stain noncovalent protein interactions. RESULTS: Maximal labeling was attained within 1 h with 1 mM CBQCA. Linear fluorescence response occurred between 8 x 10(4) and 1 x 10(6) proteins/microsphere. CBQCA staining did not disturb noncovalent protein interactions. CONCLUSIONS: We have developed methods using CBQCA and flow cytometry to quickly and simply quantify the amount of protein on the surface of a microsphere. Importantly, this approach could be extended to other formats (e.g., chips). Further, because it does not disturb noncovalent protein-protein interactions, it may be possible to use this approach to detect protein interactions without the use of purified prestained probes. PMID- 15812851 TI - Resolution of overlapping capillary electrophoresis peaks by using chemometric analysis: quantification of the components in compound reserpine tablets. AB - The application of multivariate curve resolution with alternating least squares (MCR-ALS) methods to second-order data from capillary electrophoresis with diode array detector (CE-DAD) is reported. Initial qualitative solutions obtained by evolving factor analysis (EFA) and pure-variable detection method can be further optimized by a simultaneous analysis of multiple electrophoresis run data with ALS regression. While unknown samples are analyzed simultaneously against the corresponding standards in different composition ratios, the exact amounts of common components in different CE runs can be determined by the traditional calibration curve method, and quantification can thus be achieved. The above methods are applied to the determination of the components in compound reserpine tablets in overlapping peaks from CE. The quantification results are compared with those of the first derivative of the electropherogram method and artificial neural network (ANN) method. PMID- 15812852 TI - Rapid and quantitative cyclization of multiple peptide loops onto synthetic scaffolds for structural mimicry of protein surfaces. PMID- 15812853 TI - A novel homogenous assay for topoisomerase II action and inhibition. AB - Topoisomerase II is the only enzyme able to cleave and religate double-stranded DNA; this makes it essential for many vital functions during normal cell growth. Increased expression of topoisomerase II is a common occurrence in neoplasia, and different topoisomerase II inhibitors have indeed been proven to be powerful anticancer drugs. For this reason, the topoisomerase II catalytic cycle has attracted strong interest, but only a few techniques contributing to studies in this field have emerged. All of the currently used conventional methods to elucidate the action and inhibition of topoisomerase II require separation steps and are therefore unsatisfactory in terms of sensitivity, speed, and throughput. Here, for the first time, we present an assay that works in homogenous solution. The assay is based on dual-color fluorescence cross-correlation spectroscopy (DC FCCS) and allows monitoring of topoisomerase II action and, especially, detection and discrimination of different topoisomerase II inhibitor classes. The effectiveness of our new assay was confirmed by measuring the effects of a catalytic inhibitor (novobiocin) and a topoisomerase poison (m-AMSA) with bacteriophage T4 topoisomerase as a model system, thus showing the strategy to be easy, fast, and extremely sensitive. Further development of the DC-FCCS-based assay and subsequent application in high-throughput drug screening of new anticancer drugs is proposed and discussed. PMID- 15812854 TI - Functional analysis of the aureothin iterative type I polyketide synthase. AB - The modular-type polyketide synthase (PKS) that is involved in aureothin (aur) biosynthesis represents one of the first examples in which a single PKS module (AurA) is used in an iterative fashion. Here we report on the heterologous expression of an engineered AurAB fusion protein that unequivocally proves the iterative nature of AurA. In addition, point mutations reveal that aur PKS module 4 participates in polyketide biosynthesis despite its aberrant acyltransferase domain. PMID- 15812855 TI - 2'/3'-O-peptidyl adenosine as a general base catalyst of its own external peptidyl transfer: implications for the ribosome catalytic mechanism. PMID- 15812856 TI - Deamination of adenosine by extracts of Penicillium politans NRC-510. AB - Cell-free extracts of nitrate-grown Penicillium politans NRC-510 could catalyze the hydrolytic deamination of adenosine to inosine maximally at pH 6.0 and 45 degrees C. However the same extracts could not catalyze the N-glycosidic bond cleavage of adenosine at pH 4.0, 6.0 and 8.0. Incubation of the extracts at 55 degrees C for 30 minutes caused about 31% loss in activity whereas incubation of the extracts at 60 degrees C for 15 minutes caused a complete loss of enzyme activity. Results indicated the absence of the involvement of sulfhydryl groups in the catalytic site of adenosine deaminase. The enzyme is inhibited by ethylene diamine tetraacetate indicating that adenosine deaminase is a metalloenzyme. MnCl2 and MgCl2 had a remarkable activating effect, whereas HgCl2, CaCl2 and ZnSO4 showed an inhibitory effect on enzyme activity. Dialyzing the extracts for 24 hours significantly increase deaminase activity by about 33%. The apparent K(m) value was calculated for adenosine and found to be 3.63 x 10(-3) M, which indicates high affinity of adenosine deaminase for its substrate adenosine. PMID- 15812857 TI - The oxidative stress response of the yeast Candida intermedia to copper, zinc, and selenium exposure. AB - The yeast Candida intermedia, as a model organism, was used to examine the links between the metal ions exposure, reactive oxygen species generation and oxidative stress response. To estimate intracellular peroxide and superoxide levels, the fluorescence indicators dihydrorhodamine 123 and dihydroethidium were used, respectively. Antioxidant defence systems were investigated by measuring the activity of catalase, glutathione peroxidase and superoxide dismutase and the content of reduced glutathione. Altered superoxide, peroxide, glutathione levels, and the catalase activity were perceived after the treatment with copper. In the samples treated with selenium and zinc the altered peroxide and superoxide levels, and the glutathione peroxidase activity were perceived. The results indicate that the tolerance of the yeast C. intermedia to different metal ions was correlated with the reactive oxygen species generation in the cells and with the efficiency of antioxidative defence systems. PMID- 15812858 TI - Sensitivity of different zygomycetes to the Penicillium chrysogenum antifungal protein (PAF). AB - The effects of the Penicillium. chrysogenum low molecular mass antifungal protein (PAF) on selected fungal species belonging in the Zygomycetes were investigated. A total of 21 fungal isolates from 15 different genera (Absidia, Actinomucor, Cokeromyces, Gilbertella, Micromucor, Mortierella, Mucor, Mycotypha, Rhizomucor, Rhizopus, Saksenaea, Syncephalastrum, Thamnostylum, Umbelopsis and Zygorhynchus) were tested. The inhibitory potentials of PAF at 50 microg ml(-1) on the germination of the sporangiospores and at 6.25-50 microg ml(-1) on the hyphal extension were examined on different culture media. From among the fungi regarded as opportunistic human and/or animal pathogens, PAF exhibited inhibitory effects against Absidia, Mortierella, Rhizomucor and Rhizopus species. PMID- 15812859 TI - Interference of laccase in determination of cellobiose dehydrogenase activity of Pleurotus ostreatus (Florida) using dichlorophenol indophenol as the electron acceptor. AB - Pleurotus ostreatus (Florida), ITCC 3308 produces approximately 9.0 U/ml extracellular cellobiose dehydrogenase (CDH) in cellulose medium after 7 days of growth. However, no activity could be detected if the assay was done with cellobiose as the substrate and 2,6-dichlorophenol indophenol (DPIP) as the electron acceptor in absence of any laccase inhibitor. Kinetic study showed that V(max)/K(m) value was very high for rDPIP (reduced 2,6-dichlrophenol indophenol) oxidation by laccase. Oxygen consumption rate of rDPIP oxidation by the enzyme was found to be highest among all the tested substrates. The present study indicated that rDPIP was a good substrate for laccase. Therefore, caution is needed to measure CDH activity by monitoring DPIP reduction in a system where laccase is likely to be present. PMID- 15812860 TI - Properties of a thermostable extracellular lipase from Bacillus megaterium AKG-1. AB - An extracellular lipase isolated from Bacillus megaterium AKG-1 had an optimum activity at 55 degrees C/pH 7.0. It retained 100% activity at 50 degrees C for 30 min with a half life of 30 min at 70 degrees C. A 20-70% increase in lipase activity was observed in presence of acetone (20% v/v), DMSO (20% v/v) and isopropanol (10% v/v). The enzyme activity was 92, 98 and 107% after 24 h, on treatment with 10% (v/v) acetone, benzene and isopropanol respectively. Deoxycholic acid, sodium deoxycholate, lithocholic acid, rhamnolipid, Brij 52 and cholic acid stimulated the lipase activity by 76, 36, 24, 24, 23.6 and 13%, respectively. Addition of reducing agents like sodium sulphite, sodium metabisulphite and L-cysteine-HCl, at 10 mM concentration stimulated lipase activity by 127, 146 and 150% respectively. The lipase appeared to show enantioselectivity in hydrolyzing racemic 3-acetoxy-beta-lactam as it hydrolyzed only the (+) enantiomer. PMID- 15812861 TI - Delta-aminolevulinic acid biosynthesis in Ustilago maydis. AB - A biosynthetic precursor of tetrapyrroles, delta-aminolevulinic acid (ALA), can be formed via two pathways: enzymatic condensation of glycine and succinyl-CoA by ALA synthase in animal mitochondria and some fungi, and the C5 pathway converting glutamate to ALA in plants, algae, archaea, and most bacteria. The two pathways are distinguishable using specifically radiolabeled compounds. The C1 of glutamate is lost during conversion to succinate in the TCA cycle, and the C2 of glycine is lost during conversion to acetyl-CoA on the way to glutamate. Desalted high-speed supernatants of Ustilago maydis sporidia extracts were assayed using specifically radiolabeled substrates. A significant amount of radiolabel was incorporated into ALA from 2-[14C]glycine. No radiolabel was incorporated into ALA from 1-[14C]glutamate. These results indicate that the basidiomycete yeast, Ustilago maydis, has active ALA synthase. PMID- 15812862 TI - Regulation of pyrimidine biosynthesis in Acidovorax delafieldii. AB - The regulation of de novo pyrimidine biosynthesis in the nutritionally versatile bacterium Acidovorax delafieldii ATCC 17505T was influenced by carbon source and pyrimidine supplementation. Uracil supplementation of succinate-grown A. delafieldii ATCC 17505T cells produced a greater decrease in the de novo pyrimidine biosynthetic pathway enzyme activities than did orotic acid supplementation. The presence of orotic acid or uracil in the medium of the glucose-grown wild type cells generally increased the pyrimidine biosynthetic enzyme activities. After the pyrimidine limitation of an A. delafieldii orotate phosphoribosyltransferase mutant strain, the pyrimidine biosynthetic pathway enzyme activities in the glucose-grown mutant cells were more highly derepressed than in the succinate-grown mutant cells. PMID- 15812863 TI - Role of white rot fungus Funalia trogii in detoxification of textile dyes. AB - Toxic and genotoxic effects of the textile dyes on organisms suggest the need for remediation of dyes before discharging them into the environment. For this reason, the ability of Funalia trogii pellets to detoxify textile dyes was investigated and evaluated. Although, textile dyes are toxic substances for many microorganisms, the pellets were able to decolorize and detoxify the azo dyes used. Astrazon Blue and Red dyes inhibit growth of F. trogii and S. aureus on solid medium in a concentration dependent manner. The toxicity of these dyes on a fungus, F. trogii and a bacterium, S. aureus was significantly decreased after pretreatment with fungal pellets. PMID- 15812864 TI - Malassezin, a novel agonist of the aryl hydrocarbon receptor from the yeast Malassezia furfur, induces apoptosis in primary human melanocytes. AB - Pityriasis versicolor is the most common skin mycosis in humans worldwide. Yeasts of the genus Malassezia, particularly M. furfur, a saprophyte occurring widely on human skin, are generally regarded as the causative agents. Pityriasis versicolor is often accompanied by a long-lasting depigmentation that persists even after successful antimycotic therapy. M. furfur is able to convert tryptophan into a variety of indole alkaloids, some of them showing biological properties that correlate well with certain clinical features of pityriasis versicolor. This suggests a possible role for these compounds in the depigmentation process. We now report that human melanocytes undergo apoptosis when exposed to the crude mixture of tryptophan metabolites from M. furfur. The active compound was identified as malassezin, previously isolated by us from the same source and characterized as an agonist of the aryl hydrocarbon (Ah) receptor. The compound could, therefore, contribute to the marked depigmentation observed during the course of pityriasis versicolor. PMID- 15812865 TI - Investigation of molecular beacon aptamer-based bioassay for platelet-derived growth factor detection. AB - This report describes studies on the use of a molecular-beacon aptamer (MBA) as a synthetic high-affinity DNA probe that exhibits fluorescence resonance energy transfer (FRET) in response to a specific protein biomarker, platelet-derived growth factor (PDGF). As a step toward the application of the MBA in a fluorescence-based assay for biological specimens, we examined the influence of certain physical and chemical parameters of incubation that would affect DNA conformation and DNA-backbone modification, and thus improve nuclease resistance. This bioassay is compatible with pH, temperature, and monovalent cation levels typically encountered in biological samples, and phosphorothioate backbone modified MBA is able to exhibit specific FRET. With minimal sample processing and without assay optimization, the MBA is able to detect as little as 10 ng PDGF per mug of serum proteins from cell-culture media. We also show that different sets of known fluorophore-quencher pairs can be successfully used in the MBA for sensitive detection of the PDGF target. It should, therefore, be possible to develop multiplex bioassays that monitor either quenching or enhancement for the simultaneous detection of several biomarkers by using MBAs created from high affinity DNA ligands for the desired protein targets. Interestingly, we observed that, with a DNA ligand with multiple binding sites for a standard multimeric protein target, the FRET bioassay could be accomplished by using a mixture of two individually labeled DNAs-one carrying the fluorophore and the other with the matching quencher. This observation has significant implications in the future design of more selective DNA-based FRET bioassays that use more than one ligand for the same protein target. PMID- 15812867 TI - Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus. AB - Ethanolic extracts and some fractions from 10 Indian medicinal plants, known for antibacterial activity, were investigated for their ability to inhibit clinical isolates of beta-lactamase producing methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA). Synergistic interaction of plant extracts with certain antibiotics was also evaluated. The MRSA test strains were found to be multi-drug resistant and also exhibited high level of resistance to common beta-lactam antibiotics. These strains produced beta-lactamases, which hydrolyze one or other beta-lactam antibiotics, tested. The extract of the plants from Camellia sinensis (leaves), Delonix regia (flowers), Holarrhena antidysenterica (bark), Lawsonia inermis (leaves), Punica granatum (rind), Terminalia chebula (fruits) and Terminalia belerica (fruits) showed a broad spectrum of antibacterial activity with an inhibition zone size of 11 mm to 27 mm, against all the test bacteria. The extracts from the leaves of Ocimum sanctum showed better activity against the three MRSA strains. On the other hand, extracts from Allium sativum (bulb) and Citrus sinensis (rind) exhibited little or no activity, against MRSA strains. The antibacterial potency of crude extracts was determined in terms of minimum inhibitory concentration (MIC) by the tube dilution method. MIC values, of the plant extracts, ranged from 1.3 to 8.2 mg/ml, against the test bacteria. Further, the extracts from Punica granatum and Delonix regia were fractionated in benzene, acetone and methanol. Antibacterial activity was observed in acetone as well as in the methanol fractions. In vitro synergistic interaction of crude extracts from Camellia sinensis, Lawsonia inermis, Punica granatum, Terminalia chebula and Terminalia belerica was detected with tetracycline. Moreover, the extract from Camellia sinensis also showed synergism with ampicillin.TLC of the above extracts revealed the presence of major phytocompounds, like alkaloids, glycosides, flavonoids, phenols and saponins. TLC-bioautography indicated phenols and flavonoids as major active compounds. PMID- 15812868 TI - Two faces of a biomimetic non-heme HO-Fe(v)=O oxidant: olefin epoxidation versus cis-dihydroxylation. PMID- 15812869 TI - Metal-induced chiral folding of depsipeptide dendrimers. AB - The synthesis and metal complexation of chiral depsipeptide dendrimers 3 and 7 containing an ethylenediaminetetraacetic acid (EDTA) ester-derived core is reported. The EDTA ester cavity of these dendrimers selectively complexes Zn(2+) and Cu(2+) ions leading to diastereoselective folding. To elucidate the coordination motif in the resulting "foldamers" of 3-ZnCl(2), 7-ZnCl(2), 3 CuCl(2), and 7-CuCl(2), the coordination behavior of the tetramethyl ester of EDTA (8) has been investigated as a model case. The corresponding complexes 8 ZnCl(2) and 8-CuCl(2) have been structurally characterized by (1)H NMR spectroscopy and X-ray analysis. The complexes involve the inherently chiral octahedral cis-alpha coordination motif, in which 8 serves as a tetradentate ligand. In the case of the Zn(II) complex 8-ZnCl(2), both Deltacis alpha(S,S,lambda) and Lambdacis-alpha(R,R,lambda) stereoisomers were found in the unit cell. For the Cu(II) complex 8-CuCl(2), only one stereoisomer, namely Deltacis-alpha(S,S,lambda) was found in the crystal under investigation. (1)H NMR spectroscopy has shown that the same coordination motif is diastereoselectively formed in the chiral Zn(2+) dendrimers 3-ZnCl(2) and 7-ZnCl(2). Likewise, the calculated CD spectrum of the Deltacis-alpha(S,S,lambda) stereoisomer of the model complex 8-CuCl(2) shows good agreement with the experimental spectrum of the Cu(II) dendrimers 3-CuCl(2) and 7-CuCl(2), allowing assignment of the absolute configurations of the preferred foldamers as Lambdacis-alpha(R,R,lambda) for 3-CuCl(2) and Deltacis-alpha(S,S,lambda) for 7-CuCl(2). This work represents the first example of metal-complexation-mediated diastereoselective folding of chiral dendrimers with known absolute configuration. PMID- 15812870 TI - Fluorescent probes for sensing processes in polymers. AB - Fluorescence spectroscopy is an important analytical technique that has been widely used in a variety of applications, such as biomedicine, biology, and science of materials, because it presents some properties which makes it unique, that is, extraordinary sensitivity and selectivity, short delay time (<10(-9) s), and it is neither invasive nor destructive, so it can be used for in situ measurements. Generally, intrinsic fluorescence of many materials, like polymers, is unspecific so it is not useful to analyse their properties or to be correlated to changes in their microenvironment. The incorporation of additives with fluorescent groups would be necessary. When the fluorescence emission of these molecules is sensitive to changes of properties, such as polarity, fluidity, order, molecular mobility, pH, or electric potential, they can be used for detecting such changes in their microenvironment, and they are called fluorescent probes. As long as these probes can follow processes of practical interest, they can be employed as sensors, if the information given by the measure of fluorescence adequately reflects the changes in the system. In addition, a sensor must fulfil some other requirements in order to make them of practical use, the most important being that the material support in which the sensor molecule is inserted. This support should permit a rapid detection of the process and should allow easy processing in a variety of forms. Polymers are well-known systems in which estimation of local parameters are possible by means of fluorimetric techniques. It allows the study of dynamic processes of interest, such as polymerization kinetics and mechanisms, thermal transitions, photodegradation, swelling morphology changes, and so forth. PMID- 15812871 TI - Strained, stable 2-aza-1-phosphabicyclo[n.1.0]alkane and -alkene Fe(CO)4 complexes with dynamic phosphinidene behavior. AB - The synthesis of highly strained bicyclic phosphirane and phosphirene iron tetracarbonyl complexes, that is, complexes with 2-aza-1 phosphabicyclo[n.1.0]alkanes and -alkenes (n = 3-5), is explored by using intramolecular cycloaddition of an in situ generated electrophilic phosphinidene complex, [R(iPr)NP=Fe(CO)(4)], to its C=C- and C[triple chemical bond]C containing R substituent. Saturated bicyclic complexes 7 a-c with n = 4-2 are remarkably stable, as illustrated by the X-ray crystal structure for 7 b (n=3), yet all readily undergo retroaddition to react with phenylacetylene. Shuttling of the phosphinidene iron complex between two equivalent C=C groups is demonstrated for a 1-butene-substituted 2-aza-1-phosphabicyclo[3.1.0]hexane by selective (1)H NMR magnetization transfer from the phosphirane protons to the olefinic protons. Even the more strained unsaturated bicycles 17 a,b (n = 4,3) are surprisingly stable as illustrated by the X-ray crystal structure for 17 a (n = 4), but the smaller phosphabicyclo[3.1.0]hex-5-ene (17 c, n = 2) dimerizes to tricyclic 19 with a unique ten-membered heterocyclic ring; an X-ray crystal structure is reported. Like their saturated analogues also the bicyclic phosphirenes readily undergo retroaddition as shown by the reaction of their phosphinidene iron moiety with phenylacetylene. PMID- 15812872 TI - Molecular conformation and packing of peptide beta hairpins in the solid state: structures of two synthetic octapeptides containing 1-aminocycloalkane-1 carboxylic acid residues at the i+2 position of the beta turn. AB - Peptide beta-hairpin formation is facilitated by centrally positioned D-Pro-Xxx segments. The synthetic peptides Boc-Leu-Phe-Val-D-Pro-Ac(6)c-Leu-Phe-Val-OMe (1) and Boc-Leu-Phe-Val-D-Pro-Ac(8)c-Leu-Phe-Val-OMe (2) were synthesized in order to explore the role of bulky 1-aminocycloalkane-1-carboxylic acid residues (Ac(n)c, where n is the number of carbon atoms in the ring), at the i+2 position of the nucleating beta turn in peptide beta hairpins. Peptides 1 and 2 crystallize in the monoclinic space group P2(1) with two molecules in the asymmetric unit. The crystal structures of 1 and 2 provide conformational parameters for four peptide hairpin molecules. In all cases, the central segments adopts a type II' beta-turn conformation, and three of the four possible cross-strand hydrogen bonds are observed. Fraying of the hairpins at the termini is accompanied by the observation of NHpi interaction between the Leu(1)NH group and Phe(7) aromatic group. Cross strand stabilizing interactions between the facing residues Phe(2) and Phe(7) are suggested by the observed orientation of aromatic rings. Anomalous far-UV CD spectra observed in solution suggest that close proximity of the Phe rings is maintained even in isolated molecules. In both peptides 1 and 2, the asymmetric unit consists of approximately orthogonal hairpins, precluding the formation of a planar beta-sheet arrangement in the solid state. Solvent molecules, one dioxane and one water in 1, three water molecules in 2, mediate peptide association. A comparison of molecular conformation and packing motifs in available beta-hairpin structures permits delineation of common features. The crystal structures of beta-hairpin peptides provide a means of visualizing different modes of beta-sheet packing, which may be relevant in developing models for aggregates of polypeptides implicated in disease situations. PMID- 15812873 TI - Diversity oriented synthesis of hispanane-like terpene derivatives from (R)-(+) sclareolide. AB - (R)-(+)-Sclareolide 1 has been used as a starting material to develop a diversity oriented methodology to access hispanane 28 a, and hispanane-like derivatives 27 b-27 e. This methodology is based on the intramolecular Friedel-Crafts acylation of the corresponding 12-desoxylabdanoic-like acids 27, for the construction of the cycloheptane ring which is characteristic of the hispananes. Acids 27 are obtained from alcohols 20, available by addition of the lithium or magnesium reagents to amide 12 (followed by Luche reduction), or to aldehyde 21. This sequence has resulted in the preparation of hispanane framework 27 a. The versatility of this methodology therefore allows a structural diversity oriented synthesis, since it allows the access to a wide variety of hispanane-like derivatives. PMID- 15812874 TI - Heteroditopic ligand accommodating a fused phenanthroline and a schiff base cavity as molecular spacer in the study of electron and energy transfer. AB - The synthesis and characterisation of the heteroditopic ligand N,N'-bis(3,5-di tert-butylsalicylidene)-5,6-(1,10-phenanthroline)diamine (DPSalH(2)) bearing a phenanthroline and a bis(salicylidene)diimine cavity are reported. This versatile ligand combines two of the most widely used ligands in coordination chemistry. Sequential metallation of the phenanthroline end with Ru(II) and the salophenic cavity with Cu(II) is described. Electrochemical behaviour of the supramolecular complexes [Ru(bpy)(2)(DPSalH(2))](2+) and [Ru(bpy)(2)(DPSalCu)](2+) are analysed in connection with UV/Vis and EPR spectroscopy. The data for the one-electron reduced species and the singly oxidised species of the binuclear Ru(II)-Cu(II) complex confirmed the formation of metalloradical complexes. Density functional calculations on the free ligand and the copper-only complex indicate in both cases that the HOMOs and LUMOs are developed on the Schiff base cavity with minor contributions on the bipyridine end. These findings support a bichromophoric character for our ruthenium complexes in the ground state, a necessary condition in the design of supramolecular systems for the study of electron transfer. Photophysical studies indicate fast quenching of the triplet excited state in both complexes, which suggests strong intercomponent excited-state interactions. Evidence is presented that this quenching is due to intramolecular electron transfer, at least in the case of [Ru(bpy)(2)(DPSalH(2))](2+), for which a charge separated state with a remarkable lifetime of about 30 mus was observed. PMID- 15812875 TI - Dichlororuthenium(IV) complex of meso-tetrakis(2,6-dichlorophenyl)porphyrin: active and robust catalyst for highly selective oxidation of arenes, unsaturated steroids, and electron-deficient alkenes by using 2,6-dichloropyridine N-oxide. AB - [Ru(IV)(2,6-Cl2tpp)Cl2], prepared in 90 % yield from the reaction of [Ru(VI)(2,6 Cl2tpp)O2] with Me3SiCl and structurally characterized by X-ray crystallography, is markedly superior to [Ru(IV)(tmp)Cl2], [Ru(IV)(ttp)Cl2], and [Ru(II)(por)(CO)] (por=2,6-Cl2tpp, F20-tpp, F28-tpp) as a catalyst for alkene epoxidation with 2,6 Cl2pyNO (2,6-Cl2tpp=meso-tetrakis(2,6-dichlorophenyl)porphyrinato dianion; tmp=meso-tetramesitylporphyrinato dianion; ttp=meso-tetrakis(p-tolyl)porphyrinato dianion; F20-tpp=meso-tetrakis(pentafluorophenyl)porphyrinato dianion; F28 tpp=2,3,7,8,12,13,17,18-octafluoro-5,10,15,20 tetrakis(pentafluorophenyl)porphyrinato dianion). The "[Ru(IV)(2,6 Cl2tpp)Cl2]+2,6-Cl2pyNO" protocol oxidized, under acid-free conditions, a wide variety of hydrocarbons including 1) cycloalkenes, conjugated enynes, electron deficient alkenes (to afford epoxides), 2) arenes (to afford quinones), and 3) Delta5-unsaturated steroids, Delta4-3-ketosteroids, and estratetraene derivatives (to afford epoxide/ketone derivatives of steroids) in up to 99 % product yield within several hours with up to 100 % substrate conversion and excellent regio- or diastereoselectivity. Catalyst [Ru(IV)(2,6-Cl2tpp)Cl2] is remarkably active and robust toward the above oxidation reactions, and turnover numbers of up to 6.4x10(3), 2.0x10(4), and 1.6x10(4) were obtained for the oxidation of alpha,beta unsaturated ketones, arenes, and Delta5-unsaturated steroids, respectively. PMID- 15812876 TI - Control of helix sense by composition of chiral-achiral copolymers of N propargylbenzamides. AB - N-Propargylbenzamides 1-7 were polymerized with (nbd)Rh(+)[eta(6)-C(6)H(5)B( )(C(6)H(5))(3)] to afford polymers with moderate molecular weights (M(n) = 26,000 51,000) in good yields. The (1)H NMR spectra demonstrated that the polymers have fairly stereoregular structures (81-88 % cis). The optically active polymers, poly(1) and poly(2), were proven by their intense CD signals and large optical rotations to adopt a stable helical conformation with an excess of one-handed screw sense when heated in CHCl(3) or toluene. The sign of Cotton effect could be controlled by varying the content in the copolymers of either chiral bulky 1 and achiral nonbulky 3, or chiral nonbulky 2 and achiral bulky 7. The smaller the pendant group in the copolymerization of achiral monomers with 1, the more easily did the preferential helical sense change with the copolymer composition. However, the copolymers of chiral nonbulky 2 and achiral nonbulky 3 did not change the helical sense, irrespective of the composition. The free energy differences between the plus and minus helical states, as well as the excess free energy of the helix reversal, of those chiral-achiral random copolymers were estimated by applying a modified Ising model. PMID- 15812877 TI - Use of postmenopausal hormone therapy since the Women's Health Initiative findings. AB - PURPOSE: To assess how use of postmenopausal hormone therapy (PHT) has changed since the Women's Health Initiative (WHI) trial was halted early due to an excess risk of stroke and other adverse outcomes. To estimate whether use of alternative drugs to treat menopausal symptoms (e.g., selective serotonin reuptake inhibitors [SSRIs], soy) has increased. METHODS: Women were interviewed in the Slone Survey, a random-digit-dial (RDD) survey of current medication use in a representative national sample. Information was obtained on PHT including dose, route, and reason for use, and on use of alternative drugs to treat menopausal symptoms. There were 3853 women aged >or=50 years, interviewed from 1/2001 to 6/2004. RESULTS: The average weekly prevalence of PHT declined 57%, from 28% in the first half of 2002 to 12% in the first half of 2004. Use declined for conjugated estrogens (CE) and for other estrogens, taken either alone or with progestin. The decrease exceeded 50% in most strata of age, race, education, and region. The proportion of PHT users taking 0.3 mg CE did not change. Comparing prevalence in 2004 with prevalence in 2002, there was no material increase in use of black cohosh (2.0% in 2004) or soy (2.0%) and use of SSRIs was somewhat lower (8.9%). CONCLUSIONS: These population-based usage data demonstrate a large decline in PHT use among women of postmenopausal age. The proportion of CE users taking lower doses has not increased. On a population basis, millions fewer women are using PHT in 2004 than before the WHI results were published, but there has been no appreciable increase in use of alternative therapies for menopausal symptoms over the same period. PMID- 15812878 TI - Triplet-triplet energy transfer controlled by the donor-acceptor distance in rigidly held palladium-containing cofacial bisporphyrins. AB - Eleven new complexes, including mono-, heterobi-, and homobimetallic cofacial bisporphyrins, (Pd)H2DPS, (M)H2DPX, (M)H2DPB, (PdZn)DPS, (PdZn)DPX, (Pt)2DPX, (M)2DPB (M = Pd, Pt), and (Pt)P (DPS4- = 4,6-bis[5-(2,8,13,17-tetraethyl 3,7,12,18-tetramethylporphyrinyl)]dibenzothiophene tetraanion, DPX(4-) = 4,5 bis[5-(2,8,13,17-tetraethyl-3,7,12,18-tetramethylporphyrinyl)]-9,9 dimethylxanthene tetraanion, DPB4- = 1,8-bis[5-(2,8,13,17-tetraethyl-3,7,12,18 tetramethylporphyrinyl)]biphenylene tetraanion, P2- = 5-phenyl-2,8,13,17 tetraethyl-3,7,12,18-tetramethylporphyrin dianion) have been synthesized and characterized. The photophysical properties of the donor (M)P (M=Pd or Pt, P=porphyrin chromophore) and the acceptor (free base H(2)P or (Zn)P) depend on the C(meso)-C(meso) distance and the presence of a heavy atom such as Pd(II) or Pt(II). The data were compared with those for the known compounds (Pd)2DPS, (Pd)2DPX, H4DPS, H4DPX, H4DPB, (Pd)P, (Zn)P, and H(2)P. The rate constants for triplet-triplet energy transfer (k(ET)) were measured for the heterobimetallic (PdZn) and monometallic [(M)H2] derivatives (M=Pd, Pt). The fluorescence lifetimes (Deltatau(F)) of the acceptors decrease as a result of the heavy-atom effect, and vary as follows: (Pd)H2DPS<<(Pd)H2DPX approximately (Pd)H2DPB. The k(ET) values calculated according to the equation k(ET)=(1/tau(emi)-1/tau(emi) (0)), where tau(emi) (0) is the emission lifetime of the homobimetallic bisporphyrins (no ET occurs), are equal to 0, 247+/-57 and 133+/-52 s(-1) for DPS, DPX, and DPB, respectively, in the (Pd)H(2) series. These measurements allowed the range of distance over which the Dexter mechanism for T(1)-T(1) energy transfer ceases to operate to be determined. This distance is somewhere between 4.3 and 6.3 A, in agreement with our recent findings on singlet-singlet energy transfer. During the course of this study, the X-ray crystal structure for (Pd)H2DPX was obtained; triclinic (P1), a = 11.1016(1), b = 14.9868(2), c = 20.6786(3) A, alpha = 102.091(1), beta = 100.587(1), gamma = 101.817(1) degrees , V = 3199.19(7) A(3), Z = 2. PMID- 15812879 TI - [Advances in treatment of chronic hepatitis B]. PMID- 15812880 TI - [Construction and characterization of a cDNA library from human liver tissue of cirrhosis]. AB - OBJECTIVE: To construct a cDNA library from human liver tissue of cirrhosis. METHODS: The total RNA from human liver tissue of cirrhosis was extracted using Trizol method, and the mRNA was purified using mRNA purification kit. SMART technique and CDSIII/3' primer were used for first-strand cDNA synthesis. Long distance PCR was then used to synthesize the double-strand cDNA that was then digested by proteinase K and Sfi I, and was fractionated by CHOMA SPIN-400 column. The cDNA fragments longer than 0.4 kb were collected and ligated to lambdaTripl Ex2 vector. Then lambda-phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries was strictly checked by conventional titer determination. Eleven plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. RESULTS: The titers of unamplifed and amplified libraries were 1.03 x 10(6) pfu/ml and 1.36 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 97.24 % in unamplified library and 99.02 % in amplified library. The lengths of the inserts were 1.02 kb in average (36.36 % 1 approximately equals 2 kb and 63.64 % 0.5 approximately equals 1.0 kb). CONCLUSION: A high quality cDNA library from human liver tissue of cirrhosis was constructed successfully, which can be used for screening and cloning new special genes associated with the occurrence of cirrhosis. PMID- 15812881 TI - [Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi]. AB - OBJECTIVE: To inhibit HBV core antigen gene expression with plasmid-based RNAi. METHODS: The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR. RESULTS: HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR. CONCLUSION: Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application. PMID- 15812882 TI - [Inhibition of HBs-GFP fusion gene expression by RNA interference]. AB - OBJECTIVE: To develop an effective report gene system to test the effect of small interfering RNA (siRNA). METHODS: HBV S gene was fused with enhanced green fluorescent protein (EGFP) gene to form HBs-GFP and the plasmid containing HBs GFP was constructed. A vector expressing small hairpin RNA (shRNA) pAVU6 + 4sh357 was also constructed. Two plasmids were co-transfected into HepG2 cells transiently. The fluorescence of HBs-GFP was detected by fluorescence-activated cell sorting (FACS). The mRNA expression in HepG2 cells was detected by conventional RT-PCR and real-time PCR. RESULTS: siRNA inhibited the expression of HBs-GFP 72 hours post transfection. The fluorescence of HBs-GFP in HepG2 cells treated with pAVU6+4sh357 was reduced by 55.4% compared with that of controls. The HBs-GFP expression in HepG2 cells treated with pAVU6+4sh357 was reduced by 76.3% and 90% as measured with conventional RT-PCR and real-time PCR, respectively. CONCLUSION: This investigation demonstrated siRNA derived from shRNA expression vectors can inhibit the expression of HBs-GFP in HepG2 cells. PMID- 15812883 TI - [Inhibition of binding peptides on replication of duck hepatitis B virus]. AB - OBJECTIVE: To study the inhibitory effect of binding peptides on duck hepatitis B virus (DHBV) replication in duck hepatocytes. METHODS: Specific binding peptides to duck hepatitis B virus polymerase (DHBVP) were screened by phage display technology (PDT), then were sequenced and synthesized. Binding peptides were added into primary culture of duck hepatocytes infected with DHBV in vitro. DHBV DNA in the cytoplasm, cell nucleus and medium supernatant was assayed over time. RESULTS: Seven binding peptides were obtained after 3-round screening by PDT. Duck primary hepatocytes infected by DHBV were treated with above obtained binding peptides. The DHBV-DNA levels in medium supernatant and cytoplasm of duck hepatocytes treated with synthesized peptides (the 3rd and the 6th peptide) were significantly lower than those of control cells (P<0.05). CONCLUSION: Specific binding peptides to DHBVP could inhibit the replication of DHBV. PMID- 15812884 TI - [Combination of lamivudine with thymosin alpha1 in treatment of duck hepatitis B]. AB - OBJECTIVE: To explore the inhibitory effect of combination of lamivudine with thymosin alpha1 (Talpha1) on the replication of duck hepatitis B virus (DHBV). METHODS: Peking ducks of 1 d old were challenged with DHBV-positive serum and used as a duck hepatitis B model. After treated with lamivudine for three months, the ducks were randomly grouped and treated with or without Talpha1 for 8 d. Serum DHBV titrate was observed by semi-quantitative PCR, and inflammation and degeneration of hepatocytes were observed by pathology examination. RESULTS: The serum DHBV titrate was significantly reduced (4483.2+/-5193.4 compared with 9351.8+/-5059.6) after lamivudine treatment, and it was reduced more significantly(1692.2+/-589.2) after combination treatment with Talpha1. Lamivudine reduced the degeneration degree of hepatocytes (3.2+/-0.8 compared with 4.6+/-0.5) and the inflammation degree of liver (6.2+/-3.3 compared with 8.6+/-2.8). The combination treatment with Talpha1 increased liver inflammation degree (9.0+/-5.2). CONCLUSION: Both Talpha1 and lamivudine may reduce the replication of DHBV in Peking ducks and combination treatment may have the better anti-virus effect and enhance immune response in liver. PMID- 15812885 TI - [Association of genetic polymorphisms in glutathione S-transferases M1 with hepatitis beta-related hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the association of genetic polymorphisms in glutathione S-transferases(GST) M1 with hepatitis beta-related hepatocellular carcinoma (HCC). METHODS: Genomic DNA was isolated from peripheral blood of HBsAg carriers, including 91 cases of HCC, 58 liver cirrhosis(LC), 63 chronic hepatitis B(CHB), and 134 normal controls. GSTM1 genotypes were detected by multiplex PCR. RESULTS: The null genotype of GSTM1 was significantly frequent in patients with HCC compared with controls (P<0.05), but there were no significant differences in frequency of GSTM1 null genotype among patients with liver cirrhosis, chronic hepatitis B and normal controls. Subjects carrying null genotypes of GSTM1 had higher risk of developing HCC compared with those carrying positive genotype (OR=1.81.95% CI=1.05 approximately equals 3.12). CONCLUSION: The GSTM1-null genotype may be associated with an increased risk of HCC, but not of CHB and LC. PMID- 15812886 TI - [Preparation of two derivatives from icariin and investigation of their estrogen like effects]. AB - OBJECTIVE: To investigate the estrogen-like activities of icariin (ICA), icaritin (ICT) and desmethylicaritin (DICT) and their structure/activity relationships. METHODS: ICT was hydrolyzed from ICA by cellulase and then DICT was demethylated from ICT in boron tribromide and dichloromethane system. Estrogen-sensitive MCF-7 cells and T47D cells were co-incubated with different concentrations of test compounds for 6 and 9 d respectively, and the cell proliferation was measured by MTT. RESULTS: ICT and DICT both markedly enhanced cell proliferation. Compared with estradiol (10.(-9) mol/L), the proliferative effects of 10.-6 mol/L ICT and DICT on MCF-7 cells were 90.0% and 94.0% (P<0.01), respectively, and those of T47D cells were 65.6% and 50.0%. (P<0.01). But this phenomenon was not observed with ICA. Cell proliferation induced by ICT and DICT was completely antagonized by 10.(-7 )mol/L pure estrogen receptor antagonist, ICI182,780. CONCLUSION: ICT and DICT possess estrogen-like activity of enhancing proliferation in MCF-7 and T47D cells. However, ICA appears to have no estrogenicity on MCF-7 and T47D cell lines in vitro. PMID- 15812887 TI - [Inhibition of acetamide-45 on airway smooth muscle contraction induced by electric field stimulation and methacholine in vitro]. AB - OBJECTIVE: To investigate the effects of the new antiallergic agent N-(pyridin-4 yl)-(indol-3-yl) acetamide-45 (acetamide-45) on electric field stimulation (EFS) and methacholine-induced contraction of airway smooth muscle in vitro. METHODS: Contractions were induced by EFS in isolated trachea and bronchus of rats or by cumulative methacholine concentrations in isolated trachea of guinea pigs. Changes in isometric force of isolated airway smooth muscle were measured by force transducers and recorded on a multi-channel polygraph recorder. RESULTS: Acetamide-45 inhibited the contraction induced by EFS in isolated rat airway. The IC50 was 10.74 (95% CI 8.87-13.00) micromol.L(-1) and 18.83 (95% CI 14.57-24.33) micromol.L(-1) in tracheae and bronchi, respectively. Acetamide-45 also inhibited methacholine-induced contractile response of isolated guinea pig trachea in a concentration-dependent manner. At concentrations of 3, 10, 30 micromol.L(-1) acetamide-45 significantly decreased maximal contractile response of methacholine by 24.6%-43.2% and increased EC50 of methacholine by 3.1-to 21.4-fold. CONCLUSION: Acetamide-45 inhibits EFS-or methacholine-induced contraction of isolated airway smooth muscle, and these effects might be non-specific inhibition on cholinergic receptor. PMID- 15812888 TI - [Establishment of diagnostic model of cerebrospinal protein fingerprint pattern for glioma and its clinical application]. AB - OBJECTIVE: To establish the diagnostic model of cerebrospinal protein profile for gliomas by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and bioinformatics. METHODS: Seventy-five samples of cerebrospinal fluid from patients with gliomas, benign brain tumors and mild brain traumas were collected. A total of 50 samples from gliomas and non-brain tumors were divided into training sets (33 cases including 17 gliomas and 16 non brain-tumors) and testing sets (17 cases including 5 gliomas and 12 non-brain tumors). The cerebrospinal proteins bound to H4 chip were detected by SELDI-TOF MS, the profiles of cerebrospinal protein were gained and then analyzed with artificial neural network algorithm (ANN); and the diagnostic model of cerebrospinal protein profiles for differentiating gliomas from non-brain-tumors was established. Forty-seven of cerebrospinal samples of gliomas and benign brain tumors were divided into training sets (31 cases including 13 gliomas and 18 benign brain tumors) and testing sets (16 cases including 9 gliomas and 7 benign brain tumors), the diagnostic model of cerebrospinal protein profiles for differentiating gliomas from benign brain tumors was established based on the same method. The support vector machine (SVM) algorithm was also used for evaluation, both results were very similar, but the result derived from ANN was more stable than that from SVM. RESULT: The diagnostic model of cerebrospinal protein profiles for differentiating gliomas from non-brain-tumors was established and was challenged with the test set randomly, the sensitivity and specificity were 100% and 91.7%, respectively. The cerebrospinal protein profiling model for differentiating gliomas from benign brain tumors was also developed and was challenged with the test set randomly, the sensitivity and specificity were 88.9%, and 100%, respectively. CONCLUSION: The technology of SELDI-TOF MS which combined with analysis tools of bioinformatics is a novel effective method for screening and identifying tumor biomarkers of gliomas and it may provide a new approach for the clinical diagnosis of glioma. PMID- 15812889 TI - [Construction of recombinant plasmid containing mouse vasoactive intestinal peptide gene and its expression in COS-7 cells]. AB - OBJECTIVE: To construct the eukaryotic expression plasmid containing mouse vasoactive intestinal peptide(VIP) gene with biological activities. METHODS: VIP cDNA including the sequences of signal peptide was cloned from mouse thymus by RT PCR, and then inserted into the mammalian expression vector pcDNA3.1 between Hind III and EcoR I restriction sites. COS-7 cells were transfected with pcDNA3. 1-VIP using liposome, the expression of VIP was identified by Western blot and ELISA. Supernatant of transfected cell culture was added to LPS-stimulated macrophages and the TNF-alpha production in cell medium was observed by ELISA. RESULTS: The cloned VIP cDNA was confirmed by enzyme digestion and DNA sequencing. The expression of VIP was detected in the pcDNA3. 1-VIP transfected COS-7 cells by Western blot and ELISA. The VIP in culture supernatant potently inhibited TNF alpha production by LPS-induced Macrophages in vitro. CONCLUSION: The eukaryotic expression plasmid that expresses biological active murine VIP has been constructed successfully. PMID- 15812890 TI - [Expression of alpha-smooth muscle actin in kidney of diabetic rats]. AB - OBJECTIVE: To investigate the expression of alpha smooth muscle actin (alphaSMA), a marker of myofibroblast in kidney of diabetic rats and its role in pathogenesis of diabetic nephropathy. METHODS: Diabetic disease model was made on 24 Sprague Dauley (SD) rats by an single intraperitoneal injection of streptozotocin (70 mg/kg) and 24 health SD rats served as controls. Six diabetic rats and six control rats were sacrificed at 1, 2, 4, 8 weeks after injection of streptozotocin. At each time-point, the renal morphological changes were examined by histopathology. alpha SMA and collagen-IV (C-IV) expressions were studied by immunohistochemistry (SABC). The content of alpha SMA in renal cortex was detected by enzyme-linked immunosorbent assay(ELISA). RESULTS: At all time points, renal content of alpha SMA increased in diabetic rats. Compared with controls, diabetic rats had a steady increased accumulation of C-IV. Expression of C-IV was positively correlated with that of alpha SMA. CONCLUSION: At early stage of diabetes,increased alpha SMA in kidney suggests the formation of myofibroblast, which may be involved in accumulation of C-IV and contributed to diabetic nephropathy. PMID- 15812891 TI - [Application of differential display-PCR technique in fluconazole-resistance gene expression of Candida]. AB - OBJECTIVE: To investigate the application of differential display-2PCR(DD-PCR) in research on gene expression of Candida. METHODS: Resistance to fluconazole was induced in a Candida albicans isolate 435 from vagina by culturing in YEPD broth with increasing fluconazole concentration in vitro, and the resistant isolate 435 2 (MIC=128 microg/ml ) was obtained after 80 days of incubation. Comparisons between 435 and 435-2 either in fluconazole-containing medium or in drug-free medium were performed with the modified DD-PCR including amplification with long primers, silver staining, reverse dot blot and non-radiographic labeling techniques. RESULTS: Three differential displayed bands were found which showed high homology to alcohol dehydrogenase 1 (ADH1), TOP2 and CDR1, respectively. The up-regulating expression of ADH1 and CDR1 associated with fluconazole resistance was further identified by RT-PCR. CONCLUSION: The up-regulating expression of ADH1 and CDR1 was associated with fluconazole resistance in Candida albicans, ADH1 might be a candidate of novel fluconazole resistant gene. PMID- 15812892 TI - [Changes of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases]. AB - OBJECTIVE: To investigate alterations of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases. METHODS: Twenty patients with coronary heart diseases (CHD) and 20 matched control subjects were included in the study. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After cultured for 7 days, attached cells were cytochemically analyzed. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin-binding by laser scanning confocal microscope with direct fluorescent staining. EPCs proliferation and migration were measured by MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating on fibronectin coated dishes, then adherent cells were counted. RESULTS: The number of EPCs was significantly reduced in patients with CHD compared with that of age-matched control subjects (31.8+/-7.7 compared with 59.5 +/-10.6 EPCs/x 200 field; P<0.05). In addition, the functional activity of EPCs such as proliferation, migration and adhesive capacity was also impaired in patients with CHD. CONCLUSION: EPCs number and functional activity are significantly decreased in patients with CHD. PMID- 15812893 TI - [Preparation and characterization of a directly labeled mouse anti-human CD14 monoclonal antibody ZCH-2F9-FITC]. AB - OBJECTIVE: To prepare fluorescein isothiocyanate (FITC) directly conjugated to monoclonal antibody (McAb) anti-human CD14, ZCH-7-2F9 (2F9-FITC). METHODS: After generation and purification, the purity and the murine immunoglobulin subtype of the antibody were evaluated with SDS-PAGE and multicolor flow cytometry (FCM). 2F9 McAb was directly labeled with FITC through modified Marsshall's method and the positive rate of the 2F9-FITC on different types of leukemic cells were compared with the standard CD14-FITC by FCM. RESULT: A large quantity of purified 2F9 McAb was prepared. The subtype of 2F9 was murine IgG1kappa. 2F9-FITC was successfully manufactured with A295/A280 ratio of 0.44. The positive cell percentages of 2F9-FITC and CD14-FITC on the monocytes were 84.50% and 90.08%, respectively, while those on lymphocytes were only 0.52% and 1.01%. There was no significant difference between the CD14 expressions with 2F9-FITC and CD14-FITC on each type of leukemia (n=23, t=0.922, P=0.367). CONCLUSION: 2F9-FITC has been successfully prepared and it can be applied in diagnosis and differentiation of monoblastic leukemias. PMID- 15812894 TI - [Sex differences in brain stem auditory evoked potentials and P300 examination in patients with mild cognitive impairment and Alzheimer's disease]. AB - OBJECTIVE: To compare the sex differences on brain stem auditory evoked potentials (BAEP) and P300 in the elderly patients with mild cognitive impairment (MCI) and the patients with Alzheimer's disease (AD). METHODS: Thirty-eight elderly patients with MCI, 26 cases of AD and 20 health controls (HC) were examined with BAEP, P300 . Sex difference of the variables was compared inter-and intra-groups. RESULTS: Significant sex differences of BAEP were found in the latency period wave III, V of left side in the MCI group (P <0.01), in the latency period wave IV, V of left side in HC group (P<0.01), and no such differences were found in AD group. The females had longer latency period in P300 measurement than males in MCI group (P <0.01), but there were no significant differences within AD or HC groups. The males showed significant differences in wave I, II in left side between the MCI and AD groups. The males also showed significant difference in wave I, III approximate, equals V in left side, wave IV in right side between AD and HC groups, and so did the females in wave I approximate, equals V in both sides. The males had significant difference in the left wave III, IV and right wave I, IV, V between MCI and HC groups, and so did the females in right side wave I, II, V. In P300, longer latency waves were found in AD group than in MCI group. Both sexes showed significant differences in the latency of Fz, Cz, Pz between AD and HC groups, but no significant differences were found in the latency of Fz, Cz, Pz in the females between the MCI and HC groups. CONCLUSION: Sex differences were found in the examination of both BAEP and P300 in MCI group, but not in AD group. The people of same sex have different expression of BAEP and P300 among MCI, AD and HC groups, suggesting the sex difference should be considered in the differential diagnosis. PMID- 15812896 TI - [Mapping of regulatory domain of T-protein from Escherichia coli]. AB - OBJECTIVE: To map the regulatory domain of Escherichia coli T-protein. METHODS: Fragmentation cloning was employed in cloning of 11 fragments from T-protein. The regulatory activity of each fragment was determined respectively. RESULTS: The regulatory domain of T-protein was located in the C-terminal 270 amino acids, which was the same location as PDH domain. CONCLUSION: T-protein has no independent regulatory domain. PMID- 15812895 TI - [Correlation between absorption rates of beta-adrenoreceptor antagonists in rat small intestine and their molecular structures]. AB - OBJECTIVE: To study the correlation between the absorption rate constants of beta adrenoreceptor antagonists in rat small intestinal segments and their molecular structural parameters. METHODS: The net atomic charges and the molecular volumes of 11 beta-adrenoreceptor antagonists were obtained with the semiempirical self consistent field molecular orbital calculation CNDO/2 method and Mont Carlo method respectively, using the minimum energy conformation obtained from the optimization of the standard molecular geometry with the molecular mechanics MM+ method. The stepwise multiple regression analysis was used to obtain the correlation equations. RESULTS: The absorption rate constants of beta adrenoreceptor antagonists in rat jejunum or ileum were well linearly correlated with the sum of the net charges of all hydrogen atoms and the molecular volumes. The beta-adrenoreceptor antagonist with higher lipophilicity, weaker hydrogen bonding potential,and smaller molecular volume had greater absorption rate constants. CONCLUSION: The absorption rate constants of beta-adrenoreceptor antagonists in rat small intestinal segments are mainly related with their lipophilicity,hydrogen-bonding potential and molecular size. PMID- 15812897 TI - [Screening for phenylketonuria in 726,998 neonates in Zhejiang Province]. AB - OBJECTIVE: To analyze the results of screening for neonatal phenylketonuria (PKU) in Zhejiang Province. METHODS: The screening for neonatal PKU was conducted among 726,998 newborns in Zhejiang Province. Heel prick blood specimens were collected around 72 h after birth with 6 intakes of high protein milk and the specimens were dried on S and S903 filter papers. Phenylalanine (Phe) levels were determined quantitatively with Perkin Elmer Neonatal Fluorometric PKU kits. RESULTS: Among 726,998 newborns, elevated blood Phe levels were found in 152 infants. They were all recalled for serum amino acid analysis and 32 were confirmed to have PKU with 19 males and 13 females. The earliest time of confirmation was 16 d and latest was 105 d with the median of 32 d. CONCLUSION: The data shows that the detection rate of screening for neonatal phenylketonuria in Zhejiang Province was 1/22,718. PMID- 15812898 TI - [Ultrasonic diagnosis of isolated noncompaction of the ventricular myocardium in adults]. AB - OBJECTIVE: To apply echocardiography in diagnosis of isolated noncompaction of ventricular myocardium (INVM) in adults. METHODS: Six patients with INVM underwent echocardiographic examination using HP5500 ultrasound system with the frequency of 2 approximately equals 4 MHz, and the observation was focused on ventricularmyocardium and endocardium at one-third of the apex. RESULTS: All 6 patients showed typical echocardiographic images characterized by an altered structure of the ventricular myocardium with extremely thickened, hypokinetic segments consisting of two layers: a thin, compacted epicardium and an extremely thickened endocardium, resulting in an extremely thickened ventricular wall. CONCLUSION: Echocardiography should be the first choice to be applied in diagnosis of INVM, which is a distinct entity of specific cardiomyopathy in adults. PMID- 15812900 TI - UK: developments on HIV/AIDS, stigma, and discrimination. PMID- 15812899 TI - [Progress in research on apoptosis and mitochondrial permeability transition pore]. PMID- 15812901 TI - Canada: revised guidelines on management of infectious diseases and on bleach distribution issued. PMID- 15812902 TI - Abstracts of the 13th International AEK/AIO Congress of the German Cancer Society. Wurzburg, Germany, March 13-16, 2005. PMID- 15812903 TI - Thyroid carcinoma cells and Fas-mediated apoptosis. PMID- 15812905 TI - Light Stress in Plants: Mechanisms and Interactions. Proceedings of a meeting. Edinburgh, United Kingdom, 29 March-2 April 2004. PMID- 15812904 TI - Thyroid patients target of unfair practices. PMID- 15812906 TI - Proceedings of the 5th Auger Symposium. Melbourne, 13-15 August 2003. PMID- 15812907 TI - Donor pretreatment using the aerosolized prostacyclin analogue iloprost optimizes post-ischemic function of non-heart beating donor lungs. AB - BACKGROUND: Ischemia-reperfusion injury accounts for one-third of early deaths after lung transplantation. To expand the limited donor pool, lung retrieval from non-heart beating donors (NHBD) has been introduced recently. However, because of potentially deleterious effects of warm ischemia on microvascular integrity, use of NHBD lungs is limited by short tolerable time periods before preservation. After intravenous prostanoids are routinely used to ameliorate reperfusion injury, the latest evidence suggests similar efficacy of inhaled prostacyclin. Therefore, the impact of donor pretreatment with the prostacyclin analogue iloprost on postischemic NHBD lung function and preservation quality was evaluated. METHODS: Asystolic pigs (5 per group) were ventilated for 180 minutes of warm ischemia (Group 2). In Group 3, 100 microg iloprost was aerosolized during the final 30 minutes of ventilation with a novel mobile ultrasonic nebulizer. Lungs were then retrogradely preserved with Perfadex and stored for 3 hours. After left lung transplantation and contralateral lung exclusion, hemodynamics, rO2/FiO2, and dynamic compliance were monitored for 6 hours and compared with sham-operated controls (Group 1). Pulmonary edema was determined both stereologically and by wet-to-dry weight ratio (W/D). Statistics comprised analysis of variance with repeated measures and Mann-Whitney test. RESULTS: Flush preservation pressures, dynamic compliance, inspiratory pressures, and W/D were significantly superior in iloprost-treated lungs, and oxygenation and pulmonary hemodynamics were comparable between groups. Stereology revealed a trend toward lower intraalveolar edema formation in iloprost-treated lungs compared with untreated grafts. CONCLUSIONS: Alveolar deposition of Iloprost and NHBD lungs before preservation ameliorates postischemic edema and significantly improves lung compliance. This easily applicable innovation approach, which uses a mobile ultrasonic nebulizer, offers an important strategy for improvement of pulmonary preservation quality and might expand the pool of donor lungs. PMID- 15812908 TI - Lung edema formation during cold perfusion: important differences between rat and porcine lung. AB - BACKGROUND: The aim of this study was to investigate the effect of different perfusion pressures on edema formation during cold flush perfusion with the 2 most commonly used preservation solutions in clinical lung transplantation: Euro Collins and Perfadex solutions. METHODS: Isolated rat and porcine lungs were perfused for 3 minutes at 4 degrees C to 8 degrees C at a pressure of either 10, 15 or 20 mm Hg. Weight gain was recorded continuously. Weight gain per minute was calculated after the first phase of rapid weight gain was completed. RESULTS: In the rat model, perfusion pressure of 10 mm Hg resulted in a macro- and microscopically apparent edema, irrespective of the type of preservation solution. Perfusion pressures of 10, 15 and 20 mm Hg have weight gains of 100%, 150% and 350%, respectively, after 3 minutes of perfusion. The corresponding weight gain per minute was 18%, 31% and 84% of the initial weight. There were no statistically significant differences in weight gain between the different solutions at equal perfusion pressure. In the porcine model the flow was extremely low at 10 mm Hg and no weight gain was registered, whereas the weight gain per minute at 15 and 20 mm Hg was 1.0% and 2.1% of the initial weight. CONCLUSIONS: In porcine lungs, cold perfusion at 20 mm Hg gives minimal edema formulation, whereas in rat lungs the edema formation is deleterious, irrespective of the solution used. PMID- 15812909 TI - Bringing histology to the bedside. PMID- 15812910 TI - Animal models of epidermolysis bullosa--targets for gene therapy. PMID- 15812911 TI - The disposable facility: don't clean it, just replace it. PMID- 15812912 TI - Second trial raises punitive award against sunrise to 30 million dollars. PMID- 15812913 TI - Appeal board overturns decision denying Zambian admission to Canada. AB - The Immigration and Refugee Board, Appeal Division (IRB) has granted the appeal related to an HIV-positive Zambian citizen refused admission to Canada based on "excessive demand" criteria found in the Immigration and Refugee Protection Act (IRPA). The woman had made two applications for admission to Canada. At the time of the woman's initial application, she had not been diagnosed with HIV. The appeal was granted based on humanitarian and compassionate considerations existing at the time of the first application. PMID- 15812914 TI - Severance provisions of old Ontario employment standards legislation infringe Charter equality rights of persons with disabilities. AB - On 19 January 2004, the Divisional Court of the Ontario Superior Court of Justice decided that s 58(5)(c) of the Ontario Employment Standards Act (ESA) infringed the equality rights of people living with disabilities. Section 58 provides an employee "with general entitlement to severance pay upon termination of his or her employment," but subsection (5)(c) "operates to deny severance pay to employees who are terminated in circumstances in which their contract of employment has 'become impossible of performance or frustrated' by illness or injury." PMID- 15812915 TI - Criminal charges against marijuana compassion club members stayed. AB - As reported in a newspaper article, a British Columbia Provincial Court judge has stayed charges of possession of marijuana for the purpose of trafficking. Smith and Budda were charged after a January 2002 police raid on a business operating as a "compassion club." PMID- 15812916 TI - Federal Court permits contaminated-blood litigation to proceed. AB - In ongoing legal proceedings involving people infected with HIV through contaminated blood products, on 3 June 2004 Campbell J of the Federal Court permitted the plaintiffs to refile an amended statement of claim, thus allowing the litigation to proceed. As reported in a previous issue of the Review, the court had struck out their original statement of claim, ruling (among other things) that it disclosed no reasonable cause of action and constituted an abuse of process. PMID- 15812917 TI - India: Court finds government has constitutional obligation to provide ARVs to released prisoner. AB - On 5 May 2004 the High Court of Delhi upheld an HIV-positive person's fundamental right to access treatment and medicines. This is a significant decision, given the number of people in India estimated to be infected with HIV. The decision will have implications for the government's rollout of antiretroviral (ARV) therapy. PMID- 15812918 TI - India: challenge to constitutionality of criminal law banning homosexual acts dismissed. AB - On 1 September 2004 the Delhi High Court dismissed litigation seeking to quash a law making homosexual acts a punishable offence. Section 377 of the Indian Penal Code dates back to the 1860s, during the period of British colonialism. Two NGOs, one of which was an AIDS organization involved in prevention work, challenged the law after being criminally charged. PMID- 15812919 TI - Libya: foreign health-care workers sentenced to death for deliberate infection. AB - On 6 May 2004, five Bulgarian nurses and a Palestinian doctor accused of deliberately infecting 426 children with HIV were sentenced to death by the Benghazi Criminal Court in Libya. The execution is to be carried out by firing squad. The panel of five judges also sentenced a sixth Bulgarian, a doctor, to four years' imprisonment. The doctor originally faced the same charges as the others, but instead was convicted of illegally changing foreign currency. The court did not explain the modification of charges. PMID- 15812920 TI - UK: Court grants asylum to pregnant claimant, citing European Convention. AB - An HIV-positive Ghanaian woman who applied for asylum in the United Kingdom has been granted permission to remain on the grounds that returning her to Ghana would violate the rights of her unborn child pursuant to Article 3 of the European Convention on Human Rights. Article 3 states that "no one shall be subjected to torture or to inhuman or degrading treatment or punishment." PMID- 15812921 TI - Criminal law and HIV transmission or exposure: four new cases. PMID- 15812922 TI - Mexico: discharge of HIV-positive soldiers discriminatory and unconstitutional. PMID- 15812923 TI - Bangkok 2004. Not as simple as ABC: making real progress on women's rights and AIDS. AB - In a world where the subordination and abuse of women and girls is driving the HIV/AIDS epidemic, significant resources must be invested in programs that will offer women real protection against human rights abuses. This was a central theme of Joanne Csete's presentation at "Acting on rights: women, HIV/AIDS and the Barcelona Bill of Rights," a satellite meeting held on 11 July 2004 and organized by Health Canada. This article, which is based on the presentation, discusses the limitations of the ABC approach to HIV prevention; lists some of the contextual factors that feed the subordination of women; and describes some of the violations of women and girls linked to HIV/AIDS. The author calls for the establishment of large-scale, intersectoral programs addressing the structural factors that constrain the behavioural choices of women. PMID- 15812924 TI - Bangkok 2004. Young women living with HIV/AIDS have rights too: a personal testimonial. AB - Many women living with HIV/AIDS continue to be treated as second-class citizens. In this presentation prepared for "Acting on rights: women, HIV/AIDS and the Barcelona Bill of Rights," a satellite meeting held on 11 July 2004, Promise Mthembu poignantly describes the stigma, discrimination, and abuse that she has faced as a young HIV-positive woman in Africa. The presentation discusses the issues of violence, disclosure, power imbalances in relationships, reproductive rights, safer sex, and access to health care. The presentation concludes with a series of specific recommendations for helping women living with HIV/AIDS realize their rights. Because Ms Mthembu was unable to attend the meeting in person, her presentation was read by Kanjoo Mbaindjikua, who told the delegates at the meeting that she shares many of Ms Mthembu's circumstances and experiences. PMID- 15812925 TI - Bangkok 2004. A human rights crisis in need of a human rights response. AB - Although HIV/AIDS is a human rights crisis, in many countries the protection of human rights is still not a vital component of the response. By protecting human rights, it is possible to help prevent HIV and to mitigate the personal and societal impact of the epidemic. In her presentation to a plenary session of the XV International AIDS Conference in Bangkok on 16 July 2004, Irene Khan describes how discrimination and inequality fuel HIV/AIDS, and how gender inequalities and violence render women and girls particularly vulnerable. The presentation concludes by outlining a series of measures that need to be implemented to protect human rights. PMID- 15812926 TI - Bangkok 2004. Sex workers and law reform in South Africa. AB - The Sisonke movement in South Africa aims to galvanize sex workers to fight for equal rights and for improvements in their living and working conditions. This article, based on Jayne Arnott's presentation to a plenary session at the XV International AIDS Conference in Bangkok on 14 July 2004, outlines the legislation that governs the sex trade in South Africa; reviews related legal and policy developments since the end of apartheid in 1994; describes the present environment; and outlines the contribution that sex workers themselves are making to the fight for reform. PMID- 15812927 TI - Bangkok 2004. Youth: too often missing from the response. AB - Responses to HIV/AIDS need to recognize the diversity of youth and need to be inclusive of youth. In his presentation to a plenary session of the XV International AIDS Conference in Bangkok on 14 July 2004, Raoul Fransen explains why it is important to involve youth in policy-making and at all stages of program development and implementation. The presentation emphasizes the need to ensure that young people are able to make choices concerning their sexuality, and are provided with the information and support necessary to enable them to make intelligent choices. PMID- 15812928 TI - Bangkok 2004. Double discrimination: drug users living with HIV/AIDS. AB - Drug users in Russia living with HIV/AIDS are often denied basic medical and social services and are systematically excluded from antiretroviral therapy programs. In this presentation to "HIV Treatment for Drug Users--A Realistic Goal," a satellite meeting of the XIV International AIDS Conference held on 15 July 2004, Dmitry Samoilov describes the discrimination faced by people living with HIV/AIDS in general, and HIV-positive drug users in particular. The presentation includes powerful personal testimonials from people living with HIV/AIDS. The author describes the obstacles to drug users' accessing health care, and concludes with suggestions for actions that should be taken to address the problem. PMID- 15812929 TI - Bangkok 2004. Drug control, human rights, and harm reduction in the age of AIDS. AB - In many countries, HIV prevalence among people who use illicit drugs is high. Yet many governments resist implementing effective HIV prevention measures, and drug users often lack access to care, treatment, and support, including for HIV/AIDS. Growing evidence indicates the dominant prohibitionist approach to illicit drugs is ineffective--and even counterproductive, blocking or undermining measures shown to reduce harms to drug users and to communities affected by open drug scenes. The growing debate over global drug control policy could shift us collectively away from the current, failed prescriptions to a more rational, pragmatic, and health-promoting framework of harm reduction. This article by Richard Elliott is an abridged version of a paper prepared for "Human Rights at the Margins: HIV/AIDS, Prisoners, Drug Users and the Law," a satellite meeting held in Bangkok on 9 July 2004, and organized by the Canadian HIV/AIDS Legal Network and the Lawyers Collective HIV/AIDS Unit (India). The article briefly outlines the impact of these two different policy approaches, examines international law on drug control, discusses how harm reduction reflects a human rights-based approach to drugs, and assesses some strategies for reforming global policy on illicit drugs. PMID- 15812930 TI - Bangkok 2004. In Thailand, drug users have to fight for their rights. AB - Injection drug users have been overlooked in the Thai government's response to HIV/AIDS, but not in its war on drugs. They experience constant police harassment and ineffective services, and face stigma and discrimination in society and in the health-care setting. Paisan Suwannawong, of the Thai Drug Users' Network, was scheduled to speak at the Opening Ceremonies of the XV International AIDS Conference in Bangkok on 11 July 2004. But he was placed last on a very long agenda that included Thai Prime Minister Thaksin Shinawatra, United Nations Secretary-General Kofi Annan, and Miss Universe. By the time he spoke, all the dignitaries and almost all the delegates had left the hall. In this edited version of his address, Paisan provides a personal testimonial about what it is like to be a drug user living with HIV/AIDS in Thailand, outlines the problems with the government's response, and describes other obstacles to delivering treatment and other services to drug users. PMID- 15812931 TI - Bangkok 2004. The spread of harm-reduction programs in Brazil. AB - A harm-reduction law implemented in the state of Sao Paulo in 1998 paved the way for Brazil to implement an extensive network of harm-reduction programs across the country. In his plenary address to the XV International AIDS Conference in Bangkok on 16 July 2004, Luiz Paulo Teixeira outlined the history of harm reduction in Sao Paulo, described how the Brazilian government adopted harm reduction as a national policy, and put forward some recommendations for extending harm-reduction policies and addressing key issues in drug-law reform. Mr Teixeira explained that leadership from both the political sector and civil society is necessary for harm reduction to be successfully implemented. PMID- 15812932 TI - Bangkok 2004. Prisoners' health and human rights in the HIV/AIDS epidemic. AB - Prisoners exist on the margins of society, often without access to HIV prevention, care, treatment, or support. Depriving prisoners of the means to protect themselves from HIV infection, and failing to provide prisoners living with HIV with care, treatment, and support equivalent to that available in the community, offend international human rights norms. This article provides a summary of a draft paper prepared for Human Rights at the Margins: HIV/AIDS, Prisoners, Drug Users and the Law, a satellite meeting held in Bangkok on 9 July 2004, and organized by the Canadian HIV/AIDS Legal Network and the Lawyers Collective HIV/AIDS Unit (India). The full text of the final paper, including references, is available on the Legal Network's website. The article reviews some of the international laws and instruments that protect the rights of prisoners and that set out minimum standards for treatment of prisoners; outlines activities in the prison setting that place prisoners at risk for HIV; describes some of the policies and societal factors that fuel the HIV/AIDS epidemic in prisons; and proposes a series of specific actions that should be taken now to respond to this epidemic. PMID- 15812933 TI - Bangkok 2004. Current issues and concerns in HIV testing: a health and human rights approach. AB - In the rush to scale up HIV testing, partially justified by the fact that treatment is becoming more widely available, the long-held view that testing must be voluntary, and that it must be accompanied by pre- and post-test counselling, is being increasingly questioned. However, as long as stigma, discrimination, and unequal access to care and treatment continue, the individual informed decision to take an HIV test must remain an integral step in medical practice. In this article, based on her presentation at an oral abstract session of the XV International AIDS Conference in Bangkok on 14 July 2004, Sofia Gruskin describes the developments that have led some people to question voluntary testing and counselling (VTC); outlines the factors that need to be considered in analyzing whether a proposed HIV testing strategy is effective in both health and human rights terms; calls for clarity in the use of terms such as "routine testing," "opting in," and "opting out"; and provides a list of considerations that must be addressed for any scaling up of HIV testing to be successful. PMID- 15812934 TI - Bangkok 2004. Sexual minorities, violence and AIDS in Africa. PMID- 15812935 TI - Preventing colon cancer. PMID- 15812936 TI - Are you at risk for a stroke? PMID- 15812937 TI - Walking: big benefits for the brain. PMID- 15812938 TI - A new look at blood pressure medications. PMID- 15812939 TI - I've begun to experience blurry vision. Although I have not had vision problems in the past, I do have arthritis. Could my medication be causing this problem? PMID- 15812940 TI - Depression. A definable disorder. PMID- 15812941 TI - Health tips. Potassium-rich foods. PMID- 15812942 TI - A broken bone may signal more than just a break. PMID- 15812943 TI - Prescribed vitamins. Sometimes a good choice. PMID- 15812944 TI - Dementia. Delaying its onset, slowing its progression. PMID- 15812945 TI - I've been taking vitamin E for several years. It was supposed to help prevent heart disease and cancer. Now I've heard it's not safe. What's going on? PMID- 15812947 TI - The women of women alive. Negatives of negativity. PMID- 15812946 TI - [Urinary incontinence: new pharmacologic therapies]. AB - Several classes of drugs have been investigated for their efficacy in treating overactive bladder syndrome (OAB) and stress urinary incontinence (SUI). Surgery and behavioral therapies are currently the mainstay of treating SUI. However, results are also being made available about a new oral medication, Duloxetine, which appears to be clinically safe and effective for the treatment of SUI. On the other hand, a new muscarinic receptor antagonist, Solifenacin, has been shown in clinical trials to be clinically effective, safe and well tolerated for treating OAB. PMID- 15812948 TI - The bioethics agenda and the Bush second term. PMID- 15812949 TI - Human growth hormone and the measure of man. PMID- 15812950 TI - Acorns and embryos. The embryo question I. PMID- 15812951 TI - The tragedy of equality. The embryo question II. PMID- 15812952 TI - Equality reconsidered--human frailty and human dignity. The embryo question III. PMID- 15812953 TI - The crisis of everyday life. PMID- 15812954 TI - In what sense equal? PMID- 15812955 TI - I've got you under my skin: tracking technology gets personal. PMID- 15812956 TI - Prozac papers. PMID- 15812957 TI - Approved stem cell lines contaminated. PMID- 15812958 TI - Give us the drugs. PMID- 15812959 TI - The unexamined assumptions of intellectual property: adopting an evaluative approach to patenting biotechnological innovation. PMID- 15812960 TI - Conflicts and commitment obligations. PMID- 15812961 TI - Ethics and research assessing the relative roles of genes and the environment. AB - Research on the interaction of genes and the environment is revealing that many human diseases have both genetic and environmental components. Even traditional "environmental" diseases, such as infections, appear to interact with genetic components in the human host. Environmental genetics research will inevitably increase understanding of individual susceptibilities to toxic exposures in the environment and harmful side effects of medications; therefore, it has great promise for improving the prevention and treatment of human diseases. However, realizing the benefits of this research requires careful attention to ethical issues that are particularly relevant in this context. This article reviews some of the most pressing issues related to research design and methods, as well as from the application of research results (e.g., workplace genetic screening and legal toxic torts, personal medical responsibility, and the relationship between genetics and public health measures). PMID- 15812962 TI - Drinking too much, too young: trying to find an answer to the persistent habit of binge drinking among young people vexes the nation's policymakers. PMID- 15812963 TI - States and tribes: a healthy alliance. Some of the health care models states and tribes are developing for Native Americans could be used for other residents. PMID- 15812964 TI - The ethics of human reproductive cloning: when world views collide. AB - Two camps in bioethics with seemingly opposing world views have staked out conflicting positions regarding the ethics of human reproductive cloning. These camps do not appear to share common concepts or ways of reasoning through which to exchange views and come to a meeting of minds about uses of this technology. Yet analysis of their respective approaches to several issues surrounding reproductive cloning, such as where the ethical limits of individual reproductive choice lie, whether the use of this technology would violate human dignity, whether it would create risks to the resulting fetuses and children that would make its use intolerable, and whether it would challenge certain core social values, reveals that they are not wholly opposed to one another. Indeed, it displays that they hold certain beliefs, values, and concerns in common. Moreover, it indicates that the different world views that they each presuppose, while flawed in certain respects, do not collide in every respect, but can be reconciled in significant ways that provide fertile ground for agreement about several issues related to human reproductive cloning. PMID- 15812965 TI - Overfed but undernourished: not will power, but purchasing power, may determine who eats healthy foods. PMID- 15812966 TI - Ethics of genetic modifications of behavior. PMID- 15812967 TI - The inclusion of minority groups in clinical trials: problems of under representation and under reporting of data. AB - OBJECTIVE: To evaluate the representation of minority groups in randomized control trials (RCTs), and the frequency with which this information is reported. STUDY DESIGN: Reviewers collected data on the racial/ethnic composition of study samples from all RCTs published in six leading medical journals in 1999. RESULTS: Of the 280 RCTs, most (204, 71.3%) provided no information on the race/ethnicity of participants. Of the 89 U.S.-based RCTs, 50 (56.1%) reported their minority distribution. Relative to other trials, those funded by the National Institute of Health (NIH) (n = 52) were more likely to report race/ethnicity data (55.8% vs. 23.7%; x2 = 20.9, p <_0.001) and to include nonwhite participants (13.5 % vs. 12.5%; x2=22.7, p<_0.001). CONCLUSION: Minority groups are currently under represented in clinical trials. Information on the race and ethnicity of clinical trial participants is currently underreported in six leading medical journals. Reporting of minority group information was significantly better only in NIH funded trials, which also were more likely to include nonwhite participants. This suggests that mandatory reporting policies may have a positive effect on both reporting and representation. PMID- 15812968 TI - Interval training: more benefit, less fatigue. PMID- 15812969 TI - Engineering genetic injustice. AB - In their jointly written book, From Chance to Choice: Genetics and Justice, Allen Buchanan, Dan Brock, Norman Daniels and Daniel Wikler defend 'the development and deployment of genetic intervention technologies..', including genetic enhancements, against charges that they exacerbate injustice. The present paper examines some of their arguments. The first section shows that the authors confuse real societies with just societies. The second shows that without this confusion, their arguments reveal the enormous justice-impairing potential of deploying genetic enhancements in such societies as the United States. PMID- 15812970 TI - Beneficence, determinism and justice: an engagement with the argument for the genetic selection of intelligence. AB - In 2001, Julian Savulescu wrote an article entitled 'Procreative Beneficence: Why We Should Select the Best Children', in which he argued for the genetic selection of intelligence in children. That article contributes to a debate on whether genetic research on intelligence should be undertaken at all and, if so, should intelligence selection be available to potential parents. As such, the question of intelligence selection relates to wider issues concerning the genetic determinism of behavioural traits, i.e. alcoholism. This article is designed as an engagement in the intelligence selection debate using an analysis of Savulescu's arguments to raise a series of problematic issues in relation to the ethics of parental selection of intelligence. These problematic issues relate to wider assumptions that are made in order to put forward intelligence selection as a viable ethical option. Such assumptions are more generic in character, but still relate to Savulescu's article, concerning issues of genetic determinism, private allocation and inequality, and, finally, individual versus aggregate justice. The conclusion focuses on what the implications are for the question of agency, especially if intelligence selection is allowed. PMID- 15812971 TI - A Thomistic understanding of human death. AB - I investigate Thomas Aquinas's metaphysical account of human death, which is defined in terms of a rational soul separating from its material body. The question at hand concerns what criterion best determines when this separation occurs. Aquinas argues that a body has a rational soul only insofar as it is properly organised to support the soul's vegetative, sensitive, and rational capacities. According the 'higher-brain' concept of death, when a body can no longer provide the biological foundation necessary for the operation of conscious rational thought and volition, a substantial change occurs in which the rational soul departs and the body left behind is a 'humanoid animal' or a mere 'vegetable.' I argue that the separation of soul and body does not occur until the body ceases to function as a unified, integrated organism. A rational soul is not only the seat of a human being's rational capacities; it is also the principle of the body's sensitive and vegetative capacities. Since Aquinas defines a human being as a composite of soul and body, and not with merely the exercise of rational capacities, the determination of death requires incontrovertible evidence that the body has ceased all the operations that correspond to the soul's proper capacities. The evidence of this is the body's loss of its integrative organic unity and the criterion for determining when this loss occurs is the irreversible cessation of whole-brain functioning. PMID- 15812972 TI - The birth of the empirical turn in bioethics. AB - Since its origin, bioethics has attracted the collaboration of few social scientists, and social scientific methods of gathering empirical data have remained unfamiliar to ethicists. Recently, however, the clouded relations between the empirical and normative perspectives on bioethics appear to be changing. Three reasons explain why there was no easy and consistent input of empirical evidence in bioethics. Firstly, interdisciplinary dialogue runs the risk of communication problems and divergent objectives. Secondly, the social sciences were absent partners since the beginning of bioethics. Thirdly, the meta ethical distinction between 'is' and 'ought' created a 'natural' border between the disciplines. Now, bioethics tends to accommodate more empirical research. Three hypotheses explain this emergence. Firstly, dissatisfaction with a foundationalist interpretation of applied ethics created a stimulus to incorporate empirical research in bioethics. Secondly, clinical ethicists became engaged in empirical research due to their strong integration in the medical setting. Thirdly, the rise of the evidence-based paradigm had an influence on the practice of bioethics. However, a problematic relationship cannot simply and easily evolve into a perfect interaction. A new and positive climate for empirical approaches has arisen, but the original difficulties have not disappeared. PMID- 15812973 TI - The determination of the best interests in relation to childhood immunisation. AB - There are many different ethical arguments that might be advanced for and against childhood vaccinations. In this paper I explore one particular argument that focuses on the idea that such vaccinations are justifiable because they are held to be in the best interests of a particular child. Two issues arise from this idea. The first issue is how best interests are to be determined in this case. The second issue is what follows from this to justify potential interventions within the family in relation to such vaccinations. I argue that best interests must be characterised objectively in such situations and that this means that, in at least some cases, parental decision-making about vaccinating their children may be overridden. PMID- 15812974 TI - Double skills, double knowledge. PMID- 15812976 TI - [Der KZ-Arzt Josef Mengele Zur Typologie eines NS-Verbrechers]. PMID- 15812975 TI - Prescription drugs: uses and effects. Anticoagulants. PMID- 15812977 TI - Introduction to the 3Rs (refinement, reduction and replacement). PMID- 15812978 TI - Outsourcing of animal care and biomedical resources. PMID- 15812979 TI - Who needs an "Occ Doc"? The role of an occupational medicine physician in an animal research facility. PMID- 15812980 TI - Novel IACUC outreach effort to facilitate animal protocol submission and review. PMID- 15812981 TI - Planning for dark cycle interruptions. PMID- 15812982 TI - Spotlight on success. Ann Rushforth. PMID- 15812983 TI - The "nation's conscience:" assessing bioethics commissions as public forums. AB - As the fifth national bioethics commission has concluded its work and a sixth is currently underway, it is time to step back and consider appropriate measures of success. This paper argues that standard measures of commissions' influence fail to fully assess their role as public forums. From the perspective of democratic theory, a critical dimension of this role is public engagement: the ability of a commission to address the concerns of the general public, to learn how average citizens resolve moral issues in healthcare, and to monitor public opinion on the topics addressed in the commission. Such a public forum role is supported by the critical literature within bioethics, which has deemed some commissions successful, supported more generally by the history of bioethics as a reform discourse that has brought socially important values into the medical domain, and supported more generally still by the example of the great social issues commissions of the 1960s. PMID- 15812984 TI - Ethics, regulation, and biomedical research. AB - Controversy has surrounded the institutions that facilitate discussion and regulation of American biomedical research for years. Recent challenges to the legitimacy of the President's Council on Bioethics have been focused on stem cell research. These arguments represent an opportunity to reconsider the legislation under which stem cell research is regulated, as well as to consider preexisting bodies like the Recombinant DNA Advisory Committee and National Bioethics Advisory Commission. This paper proposes a Federal Life Sciences Policy Commission, a novel commission with advisory and regulatory powers that would benefit from the positive and negative lessons learned under the legislation that currently shapes the formation and institutional characteristics of advisory bodies in the United States. The Federal Life Sciences Policy Commission would have institutional independence not present in previous advisory bodies, while maintaining the tradition of broad societal representation and thoughtful discourse that has developed in the United States. PMID- 15812985 TI - Valuing risk: the ethical review of clinical trial safety. AB - Despite its mandate on minimizing harms in clinical trials, the Common Rule provides little guidance as to how IRBs should evaluate risk. The Common Rule and derivative commentaries tend to conceptualize risk review as an expert-based endeavor aimed at an objective and universal evaluation of possible harm; they also have tended to locate risk in the research activity itself rather than in the context of research. These views of risk conflict with scholarship showing that risk evaluations are socially determined even among experts, that the context of harms can influence how persons evaluate risks, and that forums that approach risk assessment as a technical endeavor bracket from discussion the numerous values that ground risk judgments. Possible reforms are proposed for clinical trial risk review that would render it more inclusive of the different types of risk encountered and more attuned to the priorities of trial subjects. PMID- 15812986 TI - Ethics education and value prioritization among members of U.S. hospital ethics committees. AB - Calls for ethics education for members of hospital ethics committees presume that the effects and benefits of such education are well-established. This is not the case. A review of the literature reveals that studies consistently have failed to uncover any significant effect of ethics education on the moral reasoning, moral competency, and/or moral development of medical professionals. The present paper discuss this negative result and describes the author's national study of the value priorities of members of hospital ethics committees. This study discovered correlations between moral decision making and factors like age and type of institution where the committee operates. The results of this study also resemble those of previous studies in finding no correlation between ethics education and moral decision making. The author concludes that there is a need for more research on the effects of nonmoral personal, societal, and institutional factors on the moral reasoning of members of hospital ethics committees. Further, in the absence of any firm empirical basis, calls for ethics education for medical professionals and ethics committee members should be rethought. PMID- 15812987 TI - Embryonic stem cell funding: California, here I come? PMID- 15812988 TI - Vulnerability, vulnerable populations, and policy. PMID- 15812989 TI - Research participant information sheets are difficult to read. AB - Written information for potential research participants should be easy to read to enable comprehension and facilitate informed choices. We conducted an analysis of the parent information sheets for 74 research studies involving children. Readability was assessed using the Flesch Reading Ease (FRE) score. The mean FRE was 53.54 (reading age of 15.88 years) and FRE did not differ based on study characteristics. These findings suggest that the reading level of research participant information sheets is above recommended levels. Researchers should routinely compute the reading ease of study information sheets and make greater efforts to improve readability of written documents for potential participants. PMID- 15812990 TI - Making amends for clinical negligence. PMID- 15812991 TI - Medicines for human use (clinical trials) regulations 2004. PMID- 15812992 TI - Donor insemination: the secret experiment. AB - This paper gives an overview of the research that has been done into people created by donor insemination (DI) (note 1), shows how the secretive way DI is carried out makes objective knowledge of their situation impossible to obtain and describes how doctors support this secrecy. It argues that DI is a social experiment whose potential justifications are implicit theories that have either been falsified or are unfalsifiable, and that consequently DI is conducted unscientifically and unethically. In conclusion, it questions the integrity of the industry and the institutions that support it, and considers where we should go from here. PMID- 15812993 TI - Characteristic tissue interaction of the diastema region in mice. AB - Rodents have a toothless diastema between the incisor and the first molar, which may contain rudimentary tooth germs. In the lower diastema region of mice at E13, the rudimentary tooth germs, which developed into the bud stage before its removal by apoptosis, was found. The immunoreactivity to tenascin was observed in the condensed mesenchyme around the normal tooth bud and was detected in only the basement membrane in the diastema bud. This result shows that the relationship between mesenchymal condensation and tooth development. The similar patterns of Msx-1 and Msx-2 expression between the tooth bud and the diastema bud show that the diastema bud may have some other genetic mechanism in the developmental arrest of the rudimentary tooth germs rather than the Msx-1 and Msx-2 expression. Strikingly, the induction of the tooth formation was possible using tissue recombination between the oral epithelium of the diastema bud and the dental mesenchyme of the molar tooth bud, which indicates the potential capability of the diastema in the tooth formation. In conclusion, it is suggested that the condensed mesenchyme may be the key to tooth development. PMID- 15812994 TI - Hungarian nurses' attitudes to euthanasia. AB - This paper reports a questionnaire survey of the attitudes of a representative sample of Hungarian nurses towards euthanasia. PMID- 15812995 TI - From practice to publication. PMID- 15812997 TI - Listening to the public. PMID- 15812996 TI - Global bioethics and European cultures. PMID- 15812998 TI - Point of care. PMID- 15812999 TI - Endoscopically assisted Caldwell-Luc procedure for removal of a foreign body from the maxillary sinus. PMID- 15813000 TI - The cloning logjam: treaty talks break down at the United Nations. PMID- 15813001 TI - The Diane Pretty case and the occasional impotence of justification in ethics. AB - Most discussions in ethics argue that a certain practice or act is morally justified, with any underlying theory taken as supporting a guide to general action by aiding discovery of the objectively and singularly right thing to do. I suggest that this oversimplifies the agent's own experience of the moral dilemma, and I take the recent English case of Diane Pretty's request for assisted suicide as an example. The law refused, despite the obvious sympathy many felt for her. This only appears paradoxical, I suggest, because too much is expected of the concept of justification, and because moral understanding of a particular case is too often reduced to the legalistic search for general justificatory reasons. The starting point should be, I conclude, a full awareness of the phrase "there but for the grace of God go I". PMID- 15813002 TI - Toward establishing a universal basic health norm. PMID- 15813003 TI - An anthropological and historical analysis of the ethos and practice of biomedical professionals in postmodern democracies. PMID- 15813004 TI - Introduction: progress towards meeting the challenges in clinical microbiology and infectious diseases. PMID- 15813005 TI - Can infants catch caries? A review of the current evidence on the infectious nature of dental caries in infants. PMID- 15813006 TI - Dental implant use in New Zealand in 2004. AB - OBJECTIVE: To investigate the characteristics of dental implant providers in New Zealand, to evaluate providers' dental implant treatment strategies, and to determine the nature of barriers to implant usage in New Zealand. DESIGN: A postal survey questionnaire was sent to all registered dentists in New Zealand (N=1590). METHOD: Data were analyzed with the SPSS statistical package. RESULTS: Replies were received from 1005 (63.2 percent) of the surveyed dentists. The majority of dentists (76.2 percent) are interested in dental implants and promote them in their practices. Implant dentistry is performed by 49.5 percent of the respondents, with the prosthodontic aspects of implant dentistry are performed by 79.5 percent of these dentists, while the surgical stage is referred to specialists in 89.0 percent of the cases. The greatest barrier to dental implant treatment is cost. Implant dentistry is most often provided when there is financial support from ACC. CONCLUSIONS: The extent of dental implant use in New Zealand is still relatively limited. Dentists understand the benefits of dental implant therapy, but the perceived expense of treatment prevents optimal utilization. PMID- 15813007 TI - Growth modification--report of an NZAO symposium. AB - The New Zealand Association of Orthodontists has, through its Education and Research Development Group, adopted a style of symposium which will continue to be offered on an occasional basis to members. These workshop meetings aim to provide worthwhile and innovative professional development for members while simultaneously exploring contemporary research on matters of clinical importance. This report describes the proceedings of such a symposium, the theme of which was growth modification, held in Napier in March 2003. Common shortcomings of research published on this topic are discussed, and limited conclusions are offered relating to the treatment of Class II and Class III malocclusions. PMID- 15813008 TI - Changes in the selection of dental students in New Zealand. PMID- 15813009 TI - [Advantages and disadvantages of image guidance system in sinus surgery]. PMID- 15813010 TI - [Protective roles of vitamin E and coenzyme Q10 in the inner ear mitochondrial DNA 4834 bp deletion mutation of rats]. AB - OBJECTIVE: To explore the protective roles of vitamin E and coenzyme Q10 in the inner ear mitochondrial DNA 4834 bp deletion mutation of rats. METHODS: Forty-six Wistar rats were randomly divided into three groups. The rats of group A (18 rats) had admitted adriamycin 1 mg/kg by intraperitoneal injection twice a week and had taken vitamin E 50 mg/kg, coenzyme Q10 10 mg/kg orally everyday for three months. Group B (18 rats) was given the same dose of adriamycin as group A and saline instead of vitamin E and coenzyme Q10. Group C (10 rats) was given Saline only. The inner ear tissue was harvested and mitochondrial DNA was amplified to identify t he 4834 bp deletion mutation by nested-primer polymerase chain reaction (nested-PCR) technique. And the serum glutathione peroxidase (GSH-PX) activity was measured. RESULTS: The incidence of mitochondrial DNA 4834 bp deletion mutation of group A was 23.08% (3/13), group B was 68.75% (11/16), and group C was 0 (0/8). The mutation incidence of group A was significantly lower than group B (Fisher's exact test, 1-sided, P = 0.018) and there was no significant difference between group A and group C (Fisher's exact test, 1-sided, P = 0.215). The activity of serum glutathione peroxidase of group A was marked higher than that of group B (adjusted t' test, 1-sided, t' = 6.474, P < 0.01). And the difference of activity of serum glutathione peroxidase between group A and group C was not significantly (adjusted t' test, 2-sided, t' = 1.920, P > 0.05). CONCLUSION: The results suggested that vitamin E and coenzyme Q10 could improve the ability of free radicals cleaning and protect the mitochondrial DNA from 4834 bp deletion mutation. PMID- 15813011 TI - [Epidermal stem cells in the tympanic membrane]. AB - OBJECTIVE: To investigate the distributions of epithelial stem cells in the tympanic membrane and the growth characteristics of cultured epithelial cells from different region of tympanic membrane, and to establish culture techniques of stem cells in tympanic membrane. METHODS: Four young rats and four adult SD rats were used to observe normal tympanic membrane. The other 28 rats were performed 2 mm size perforations in pars tensa. These animals were sacrificed at different periods after perforation. The tympanic membranes were cut in cryostat sections for immunohistochemistry of cytokeratin 19 and integrin beta1. Thirty tympanic membranes of rats were treated with Mitomycin C to damage the mucosal surface, then divided into two parts: the annulus region and center region of pars tensa, cultured in medium with high amount of epidermal growth factor and low amount of calcium. RESULTS: The immunostaining cells of cytokeratin 19 and integrin beta1 were displayed in both the handle of malleus and annular regions, but there were no staining positive cells in the intermediate region of pars tensa. The positive cells distribution had no significant difference between adult and infancy rats. In the pars flaccid, the positive cells scattered in the basal layer. The positive cells increased after perforation in the annulus and handle of malleus region, but no immunostaining cells were found at the edge of perforation. The redouble time of the culture cells from the annulus region was shorter than the center of pars tensa. The cells adherent within 1 hour formed larger and more colonies, and contained more positive cells. CONCLUSION: The epithelial stem cells in tympanic membrane were located in both the handle of malleus and annular regions, but no stem cells could be found in the intermediate region of pars tensa. The stem cells of tympanic membrane can be simply purified according to the adherent time. PMID- 15813012 TI - [Overexpression of thymosin beta4 in the cochlea of senescence-accelerated mouse]. AB - OBJECTIVE: To develop gene expression profiles of young and old senescence accelerated mouse (SAM) cochlea and identify genes responsible for aging-related hearing loss. METHODS: Gene micro-array slides containing 1101 mouse genes were hybridized to cDNA micro-arrays (Atlas Glass Array Mouse 1.0) that were synthesized using total RNAs from the cochlea of 2 mounts and 12 mounts mouse. Hybridization signals were visualized with cyanine-3 fluorescent reporter molecules, and the fluorescence intensities of the images were analyzed. Real time quantitative reverse transcription PCR (qRT-PCR) was performed to validate the micro-array results. Immunofluorescence was used to identify the located region of the protein encoding by the candidate gene in the cochlea. RESULTS: Expression of a majority of the 1101 genes represented on the micro-array slides was not altered during aging; nonetheless, changes in the expression of 3 genes were detected between young and old mouse cochlea. RT-PCR results confirmed the changes in expression of thymosin beta4 of 3 genes examined. Through the using of immunofluorescence, it was shown that thymosin beta4 was primarily located in the tectorial membrane and the supporting cells of outer hair cell. CONCLUSIONS: Using commercially available slide micro-arrays, the results show that aging of the mouse cochlea is associated with changes in patterns of gene expression. This analysis suggests that thymosin beta4 may play a role in aging-related hearing loss. These studies lay the foundation for future studies defining the genetic basis of aging-related hearing loss. PMID- 15813013 TI - [Expression profile of immune associated genes in nasal polyps]. AB - OBJECTIVE: To investigate the expression profile of immune associated genes in nasal polyps by gene chip technology and to probe into the role of correlative genes in the immune pathogenesis of nasal polyps. METHODS: Microarray analysis was used to find the expressing profile of 491 immune associated genes in nasal polyps. The total RNAs were respectively extracted from four samples of nasal polyps and inferior turbinates, and then were reversely transcribed to cDNAs with incorporation of fluorescent dUTP as the hybridization probes. The mixed probes were then hybridized with two pieces of immune associated gene chip. It was scanned by laser scanner and the acquired image was analyzed by software. RESULTS: Eighty-seven genes were differently expressed in immune associated gene profile of nasal polyps, among which 45 genes were upregulated and 42 genes were down regulated. Fifteen genes were shown differential expression in both chips with 5 upregulated genes and 10 downregulated genes. The differentially expressed genes mostly involved in cytokines and their receptors, chemokines and their receptors, adhesion molecules, leukocytes differential antigens, immune signal transduction molecules, and still included some genes about complements and their receptors, immune transcription regulatory molecules, innate immune molecules and neural immune molecules. CONCLUSIONS: The differently expressed genes in immune associated gene chips will provide clues and theoretical foundation for the pathogenesis of nasal polyps. Furthermore IL-17 may have an important role in the occurrence of nasal polyps, and the role of innate immunity and immune signal transduction molecules in the pathogenesis of nasal polyps need further researches. PMID- 15813014 TI - [Effect of immunotherapy on house dust mite allergen-specific IgG1 and igG4 antibodies in nasal secretion for patients with perennial allergic rhinitis]. AB - OBJECTIVE: To investigate the effect of immunotherapy on allergen-specific IgG1 and IgG4 in nasal secretion for patients with nasal allergy. METHODS: Nasal secretion was collected with suction from normal subjects, patients with allergic rhinitis untreated and treated with immunotherapy. Indirect, non-competitive streptavidin-biotin ELISA was applied to analyze allergen-specific IgG1 and IgG4 in nasal secretion. RESULTS: Patients with nasal allergy both treated and untreated showed a higher concentration of IgG1 and IgG4 (Z = -3.623, -3.061, P < 0.01, respectively), treated group showed higher concentration of IgG1 and IgG4 than untreated group (Z = -2.453, -3.408, P < 0.01, respectively). IgG4 antibody specific to purified allergen Der f I in the treated group was higher than normal subjects (Z = -3.518, P < 0.01), both IgG1 and IgG4 antibodies specific to purified allergen Der f II in treated group were higher than normal subjects (Z = -2.366, -2.936, P < 0.01, respectively) and than untreated group (Z = -2.366, 2.937, P < 0.01, respectively). In the IgG subclasses of IgG1 and IgG4, there was positive correlation among crude mite extract, Der f I and Der f II. There was a positive correlation between IgG1 and IgG4 for all 3 kinds of allergens. There was a positive correlation between IgG1 in the nasal secretion and IgE in the serum and between IgG4 in the nasal secretion and IgE in the serum for crude mite extract. CONCLUSIONS: Patients with nasal allergy showed higher responsiveness to natural allergen exposure and produced higher level of IgG1 and IgG4 antibodies to specific allergens. Immunotherapy increased the concentration of IgG1 and IgG4 antibody in the nasal secretion, which might function as a blocking antibody to specific allergen. PMID- 15813015 TI - [Analysis of related factors of nasosinusitis after radiotherapy for nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the incidence, clinical features and related factors of nasosinusitis after radiotherapy for nasopharyngeal carcinoma. METHODS: Five hundred and thirteen patients with nasopharyngeal carcinoma were included in the study, to observe the clinical manifestation and image changes before and after radiotherapy. The incidence and influencing factors of nasosinusitis after radiotherapy were analyzed. RESULTS: Among 513 patients, before radiotherapy, nasosinusitis was found in 51 patients (9.9%). After radiotherapy, another 401 nasosinusitis was found (401/462). The difference of incidence rate of nasosinusitis before and after radiotherapy was obvious (chi2 = 533.21, P < 0.01). The incidence rate of nasosinusitis in the end of radiotherapy, 3 months, 6 months, 12 months and 18 months after radiotherapy was 10.7% (43/401), 13.7% (55/401), 58.1% (233/401), 12.0% (48/401), 5.5% (22/401) respectively. The incidence rate of nasosinusitis after fractional radiotherapy and continuous radiotherapy was 35.7% (143/401), 64.3% (258/401) respectively. CONCLUSION: The incidence rate of nasosinusitis after radiotherapy is very high. It is influenced by the dose of radiotherapy, but it has no relation with the extension of nasopharyngeal carcinoma. PMID- 15813016 TI - [Diagnosis and management of sulcus vocalis]. AB - OBJECTIVE: To clarify the pathophysiology of sulcus vocalis and to develop a more rational approach to treatment. METHODS: Twenty-nine cases of sulcus vocalis patients were divided into three classification: Type I is a physiologic variant and no dysphonia (11 cases). Types II (sulcus vergeture, 13 cases) and III (sulcus vocalis, 5 cases) are characterized by severe dysphonia and loss of vibratory activity. Eighteen cases of dysphonia were treated by surgery and phonation training. The operations included fat injection into vocal cords (9 cases of types II and 1 cases of types III, including 1 case of types III of second operation), fat implantation into sulcus vergeture after incision (4 cases of types II and 1 cases of types III) and undermining of the mucosa and sulcus vocalis resection (4 cases of types III, including 1 case of second operation). Phonatory function and video laryngostroboscopic data were evaluated before and after surgery and phonation training treatment in 18 patients. The mean follow-up time was 15.3 months. RESULTS: Ten cases of types II had excellent results after fat injection into vocal cords (n = 6) and fat implantation into sulcus vergeture after incision (n = 4). Three cases of types II improved after fat injection into vocal cords. Three cases of type III had excellent results after sulcus vocalis resection. One case of type III had excellent results by Second operation (sulcus vocalis resection) after fat injection into vocal cord. One case of type III improved by Second operation (fat injection into vocal cords) after fat implantation into sulcus vergeture after incision. No postoperative complications were noted. CONCLUSION: Accurate classification of sulcus vocalis is important and then adapt treatment to different types. Fat implantation into sulcus vergeture to type II and sulcus vocalis resection to type III were the best choice methods. PMID- 15813017 TI - [Clinical characteristic and treatment of laryngeal scleroma]. AB - OBJECTIVE: To study the clinical manifestation and treatment of the laryngeal scleroma METHODS: Forty-three patients with laryngeal scleroma, from May, 1981 to December, 2002, with pathological diagnosed were reviewed retrospectively. The clinical features and the treatment methods of these patients were analysed. RESULTS: All 43 cases had hoarseness and 19 of them manifested dyspnea. In the larynx, the main scleromatous lesions were classified as atrophic stage in 2 cases, as granulomatous in 35 cases and as scarring in 6 cases. The preponderance lesions were located in the glottis in 13 cases, in the subglottis in 1 case, in both supraglottis and glottis in 18 cases, in both glottis and subglottis in 8 cases, and in all the three regions of the larynx in 3 cases. Twenty-four patients were treated with antibiotics, seven patients with surgery and two patients with radiotherapy. Eighteen of 24 patients who were treated with antibiotics were cured, and two of the recurrent patients were cured with a further period of antibiotics therapy. Four of the 24 patients with second or third degree of laryngeal obstruction required prophylactic tracheostomy. One patient in granulomatous stage with the second degree of laryngeal obstruction was cured with the combination of surgery and antibiotics therapy. Six patients in the scarring stage with laryngeal stenosis were cured with reconstruction surgery. One patient treated with radiotherapy recurred in the fourth year after treatment, and one patient failed to antibiotics therapy cured with radiotherapy combined with antibiotics therapy. CONCLUSIONS: Scleroma may involve the larynx and cause dysphonia and laryngeal obstruction. Antibiotics therapy is effective in most cases with laryngeal scleroma, and the long-time follow-up after treatment were necessary. Laryngeal reconstructions were necessary for the patients with cicatrical laryngeal stenosis. PMID- 15813018 TI - [Clinical pathology feature and prognostic factors of cervical lymph node metastases in hypopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the risk clinicopathological factors of primary tumor in the prediction of cervical lymph node metastases and the cervical lymph node prognostic factors in hypopharyngeal squamous cell carcinoma. METHODS: A retrospective study was carried out to review the histopathological data from 98 hypopharyngeal squamous cell carcinoma patients. The relationship between histopathological parameters and cervical lymph node metastases were evaluated by means of a univariate chi2 test and multivariate stepwise logistic regression model. And the Cox regression model was used to define possible pathological parameters of neck node affecting survival including N staging, presence of cervical lymph node metastases and extracapsular nodal spread, size and number of positive neck nodes, and levels of positive neck nodes. RESULTS: The overall 5 year survival rate of patients with hypopharyngeal carcinoma was 28.6%. In a univariate and multivariate analysis, it was confirmed that size and growth pattern of primary tumor correlated to cervical lymph node metastases. In a multivariate Cox regression analysis, the most significant prognostic factors of cervical lymph node were the size of positive neck nodes and level involved. CONCLUSIONS: Cervical lymph node metastases were one of the most significant prognostic factors of hypopharyngeal carcinoma. The identification of patients at risk for cervical lymph node metastases and the management of the neck by coping with pathological factors of cervical lymph node affecting survival are very important to improve the treatment and prognosis of hypopharyngeal carcinoma. PMID- 15813019 TI - [Salvage surgery for patients with metastatic squamous cell carcinoma involving carotid artery in head and neck]. AB - OBJECTIVE: To study the surgical approaches for metastatic squamous carcinoma involving carotid artery in head and neck. METHODS: Fifty-seven patients with metastatic squamous carcinoma involving carotid artery were analyzed retrospectively. Forty-eight were undergone peel-off tumor from carotid artery and another 9 with carotid artery resection due to being invaded in the middle layer of the carotid artery wall. None of them were received artery reconstruction. Postoperative radiotherapy was given to those who had radiation with the total dose less than 70 Gy or no radiotherapy, or gross tumor residue during operation. RESULTS: Of all the 57 patients, 53 had been death and only four survived without clinical lesion. One patient died of postoperative multi organ failure, 10 of local recurrence, 19 of regional recurrence, 14 of distance metastases, 7 of carotid artery bleeding eroded by tumor, and the remaining 2 of unknown causes. Median survival was 13 months and 1, 2, 3, 5-year overall survival rate was 44.6%, 16.2%, 12.5%, 8.3% respectively, as estimated by the Kaplan-Meier approach. The recurrent rate of gross negative and positive were 26.7% and 50.0% respectively. 2-year survival rate of those were 22.2% and 0 respectively. Recurrent rate and 2-year overall survival rate in patients with and without postoperative radiotherapy was 46.4% and 21.4%, 73.1% and 15.4% respectively. CONCLUSIONS: It is necessary to perform aggressive salvage operation for patients with metastatic squamous cell carcinoma involving carotid artery. En blok removal of the tumor and the reasonable safe margin should be achieved. Unless the middle layer of the artery wall is involved, tumor peel-off is preferred to carotid resection to reduce the mortality and cerebrovascular complication. Postoperative radiotherapy reduces the recurrent rate and improves overall survival rate potentially. PMID- 15813020 TI - [Diagnosis and surgical management of Hashimoto's disease coexistent with thyroid carcinoma]. AB - OBJECTIVE: To study clinical assessment and management of the Hashimoto's disease coexistent with thyroid carcinoma. METHODS: A retrospective analysis in 182 cases of Hashimoto's disease treated from 1982 to 1998 were undertaken. The preoperation assessment, surgical management and the complication after operation had been studied. RESULTS: Eighteen cases of thyroid carcinoma were found in total of 182 patients. The coexistent rate of the Hashimoto's disease with thyroid carcinoma was 9.9% (18/182). Preoperative diagnostic accuracy was only 33.3% (6/18). The 5-year and 10-year survival rates were 88.9% (16/18) and 81.3% (13/16), respectively. Seven received second operation, 2 of them found cancer in the specimens . The 5 years survival rates of the differentiated thyroid cancer (88.2%) was higher than the undifferentiated thyroid cancer (0), but there were no statistical difference due to the small cases (P = 0.167). Two patients had local recurrence and 3 had metastases. They had salvage surgery, which was curative. Three patients with undifferentiated thyroid cancer died of remote metastases. Six patients suffered hypothyroidism. CONCLUSIONS: The preoperation diagnosis accuracy of Hashimoto's disease coexistent with thyroid carcinoma was low. A detail case history,computed tomography (CT), fine needle aspiration (FNA) can help the disease to be diagnosed. The surgical treatment is effective on patients with Hashimoto's disease coexistent with thyroid carcinoma. PMID- 15813021 TI - [Effects of intragastric administration of thenorphine on morphine-induced behavioral sensitization in mice]. AB - AIM: To investigate the effects of intragastric administration of thenorphine (Then) on behavioral sensitization to morphine (Mor) in mice. METHODS: Locomotor activity was detected after intragastric administration of thenorphine or co administration of thenorphine with Mor in mice. Mice were induced to be behaviorally sensitive to Mor, and were given the combination of Mor and thenorphine to observe the effects of thenorphine on the development, transfer and expression of Mor-induced behavioral sensitization. RESULTS: A single intragastric administration of thenorphine (1.25 - 5.0 mg x kg(-1)) dose dependently inhibited the locomotor activity in mice (P < 0.01) and the effects of thenorphine on locomotor activity developed tolerance after repeated administration. Co-administration of thenorphine effectively inhibited Mor induced hyperactivity (P < 0.05) and the development, transfer, expression of Mor induced behavioral sensitization in mice (P < 0.05 or P < 0.01). CONCLUSION: Thenorphine was shown to suppress the central nervous system and may be effective against the abuse and addiction to opioids. PMID- 15813022 TI - [Mechanism of dracorhodin perchlorate-induced Hela cell apoptosis]. AB - AIM: To study the mechanism of dracorhodin perchlorate-induced Hela cell apoptosis. METHODS: Cell viability was measured by MTT method. Morphological changes were observed by phase contrast microscopy and Hoechst 33258 staining. DNA fragmentation was assayed by agarose gel electrophoresis. Protein expression was detected by Western blot analysis. RESULTS: Dracorhodin perchlorate induced Hela cell apoptosis. The apoptosis was partially reversed by caspase-1, -3, -8, 9 and caspase family inhibitors. Treatment of Hela cells with dracorhodin perchlorate for 12 h increased the protein expression ratio of Bax/Bcl-XL; procaspase-3, -8, ICAD and PARP were cleaved to smaller molecules. CONCLUSION: Dracorhodin perchlorate induced Hela cell death via alteration of Bax/Bcl-XL ratio and activation of caspases. PMID- 15813023 TI - [Effect of MDR1 polymorphic expression on oral disposition of cyclosporine A]. AB - AIM: To determine the relationship between C3435T mutation in exon 26 of the human multidrug resistant 1 gene and cyclosporine (CsA) pharmacokinetic (PK) parameters among healthy Chinese volunteers by nonlinear mixed effect model (NONMEM). METHODS: Twenty healthy subjects were given orally a single dose of 500 mg CsA in microemulsion solution. Blood CsA concentrations were measured with HPLC and the genotype for the C3435T polymorphism of MDR1 gene was determined with the PCR and restriction fragment length polymorphism. The results were further confirmed by sequencing. NONMEM was performed to assess the effect of genotype on CsA PK profile. RESULTS: MDR1 C3435T genotype was identified as the best predictor of CsA systemic exposure. The relative bioavailability of CsA was 40% higher in subjects who carried at least one 3435C allele compared to that of TT type individuals in the study population. CONCLUSION: The MDR1 C3435T genotype offers a potential basis of mechanism to explain inter-subject differences in CsA oral bioavailability. PMID- 15813024 TI - [Effects of capsaicin on IA and IK in cultured trigeminal ganglion neurons of rat]. AB - AIM: To investigate the effect of capsaicin on IA and IK in cultured rat trigeminal ganglion (TG) neurons. METHODS: Whole-cell patch clamp technique was used to record the IA and IK before and after capsaicin perfusion at different concentrations. RESULTS: In capsaicin-sensitive (CS) neurons, capsaicin was shown to selectively inhibit IA in dose-dependent manner, the IC50 was 0.99 micromol x L(-1). Yet capsaicin showed no inhibitory effect on IK, capsaicin (10 micromol x L(-1)) only slightly inhibited IK by 13.2%. In capsaicin-insensitive (CIS) neurons, capsaicin (1 micromol x L(-1)) showed no significant inhibitory effect on IA and IK, capsaicin (10 micromol x L(-1)) only slightly inhibited IA and IK by 16.8% and 15.3%, respectively. Neither 1 micromol x L(-1) nor 10 micromol x L( 1) capsaicin showed effect on the G-V curve of IA and IK. CONCLUSION: Capsaicin was found to selectively inhibit the IA current in CS neurons, and this effect may contribute to hyperalgesia when capsaicin was first used. PMID- 15813025 TI - [Iptakalim enhances astrocytic glutamate uptake activity]. AB - AIM: To investigate the effect of iptakalim (Ipt), a novel ATP-sensitive potassium channel opener (KATPCO), on astrocyte glutamate uptake activity. The mechanism of action involved was also examined. METHODS: Primary cultures of astrocytes were prepared from cerebral cortexes of 1 to 2 days old Sprague-Dawley rats. Cultures were preincubated with iptakalim of various concentrations with the absence or presence of 6-hydroxydopamine (6-OHDA, 50 micromol x L(-1)). Then, the cells were treated with pinacidil (10 micromol x L(-1), K+ channel opener) instead of iptakalim and pretreated with glibenclamide (20 micromol x L(-1), ATP sensitive K+ channel inhibitor) before pinacidil or iptakalim was added into the medium so that the mechanism of action of iptakalim could be analysed. Astrocyte glutamate uptake activity was determined by intracellular concentration of 3H labeled D,L-glutamate, using isotope techniques. RESULTS: Both iptakalim and pinacidil were shown to increase glutamate uptake activity of astrocytes and reverse inhibitory effect induced by 6-OHDA. But the increased and reversed effects of iptakalim or pinacidil on astrocytic glutamate uptake were found to be completely antagonized by glibenclamide. CONCLUSION: These data indicated that iptakalim can enhance astrocytic glutamate uptake activity due to its K+ channel opening properties and lessen glutamate excitotoxicity. Iptakalim may be useful for the therapy of Parkinson's disease. PMID- 15813027 TI - Isolation and identification of a new dicoumarin from the roots of Daphne genkwa. AB - AIM: To investigate the chemical constituents of the roots of Daphne genkwa. METHODS: Silica gel and Sephadex LH-20 column chromatographic techniques were employed for the isolation and purification. The structure was elucidated by a combination of spectroscopic analyses. RESULTS: A new dicoumarin named isodaphnoretin was isolated and the structure was established as 7-hydroxy-6 methoxy-4-[(2-oxo-2H-1-benzopyran-7-yl)-oxyl]-2H-1-benzopyran-2-one. CONCLUSION: Isodaphnoretin is a new compound. PMID- 15813026 TI - [Synthesis and antifungal activity of novel triazole antifungal agents]. AB - AIM: A series of triazole antifungal agents were synthesized to search for novel triazole antifungal agents with more potent activity, less toxicity and broader spectrum. METHODS: Twenty-one 1-(1H-1, 2, 4-triazolyl)-2-(2, 4-diflurophenyl)-3 (4-substituted-1-piperazinyl)-2-propanols were synthesized, on the basis of the three dimensional structure of P450 cytochrome 14alpha-sterol demethylase (CYP51) and their antifungal activities were also evaluated. RESULTS: Results of preliminary biological tests showed that most of title compounds exhibited activity against the eight common pathogenic fungi to some extent and the activities against deep fungi were higher than that against shallow fungi. In general, phenyl and pyridinyl analogues showed higher antifungal activity than that of the phenylacyl analogues. CONCLUSION: Several title compounds showed higher antifungal activities than fluconazole and terbinafine. Compound VIII-1, 4, 5 and IX-3 showed the best antifungal activity with broad antifungal spectrum and were chosen for further study. PMID- 15813028 TI - [Determination of levodropropizine and its pharmacokinetics in human plasma using LC/MS/MS]. AB - AIM: To develop a rapid and sensitive LC/MS/MS method for the analysis of levodropropizine in plasma and study the pharmacokinetics of levodropropizine in healthy Chinese volunteers. METHODS: Levodropropizine and zolmitriptan (internal standard, IS) were extracted from plasma samples and chromatographed on a C18 column and detected using a tandem mass spectrometer with a TurboIon Spray ionization interface. Quantitation was performed using multiple reaction monitoring (MRM) of the transitions of the m/z 237 --> m/z 120 for levodropropizine and m/z 288 --> m/z 58 for the IS. RESULTS: The limit of quantification of the method for levodropropizine was 0.25 microg x L(-1). The assay was linear over the concentration range from 0.25 to 500.0 microg x L(-1) and intra- and inter-day precision over this range were < 11.4% with good accuracy. CONCLUSION: The method is shown to be accurate, and suitable for clinical pharmacokinetic study of levodropropizine. PMID- 15813029 TI - [Comparison between the post-column derivatization with bromine by HPLC and the fluorometric analysis for determination of aflatoxins in medicinal herbs and plant extracts]. AB - AIM: To compare the post-column derivatization technique (IAC-PCD-HPLC) for the determination of aflatoxins B1, B2, G1 and G2 and the rapid procedure with fluorometric analysis (SFB) for the determination of total aflatoxins. METHODS: The method of post-column derivatization with bromine by HPLC consisted of extraction of the sample with MeOH-H2O (70:30) followed by clean-up of the extracts with immunoaffinity columns and finally, HPLC determination with fluorescence detection. Aflatoxins B1 and G1 were determined as their bromine derivatives, produced in an on-line post-column derivatization system. In SFB method, samples were ground and extracted with methanol-water (70:30). A portion of the extract was cleaned up by passage through a immunoaffinity column, One mL of purified extract was derivatized with a bromine reagent, and fluorescence of the solution was immediately quantified with a calibrated fluorometer containing a broad wavelength pulsed xenon light source. RESULTS: In IAC-HPLC method, the overall average recoveries for three different medicinal herbs spiked at levels of 1.3 and 2.6 ng x g(-1) of total aflatoxins ranged from 93% to 97%. The detection limit was 0. 06 microg x kg(-1) for both G2 and B2 and 0.20 microg x kg(-1) for both G1 and B1, based on a signal/noise 3:1 and the precision (within laboratory relative standard deviation) ranged from 0.8% to 1.4%. Each of aflatoxins B1, B2, G1 and G2 in 39 kind medicinal materials were determined by IAC-PCD-HPLC, and the total aflatoxins were determined by SFB. CONCLUSION: The SFB method is not the suitable method for the determination of total aflatoxins in medicinal herbs and plant extracts. PMID- 15813030 TI - [Effect of programmed humidification and temperature on drug stability]. AB - AIM: To simplify the study on the effect of relative humidity and temperature on drug stability. METHODS: The stability of penicillin potassium as a model was studied with programmed humidifying and heating. RESULTS: Results of our programmed humidifying and heating experiments are comparable to those of traditional experiment at constant humidity and temperature. CONCLUSION: Programmed humidifying and heating experiments are applicable to drug stability study. PMID- 15813031 TI - [Enhancing effect of Ulex europaeus agglutinin I modified liposomes on oral insulin absorption in mice]. AB - AIM: To investigate the enhancing effect on insulin absorption through GI. tract in mice by using the Ulex europaeus agglutinin I (UEA1) modified liposomes as the carrier. METHODS: UEA1 modified phosphatidylethanolamine (PE) was prepared by conjugating method of 1-ethyl-3-(3'-dimethylaminopropyl) carbodiimide (EDC), then the modified compound (PE-UEA1) was incorporated into the conventional liposomes of insulin to obtain UEA1 modified liposomes. The agglutination test was performed to examine the UEA1 biological activities after synthesis and modification. When liposomes were applied to healthy mice or diabetic mice at insulin dose of 350 u x kg(-1) orally, the hypoglycemic effect was investigated according to the blood glucose level determination. RESULTS: The blood glucose levels of the healthy mice reduced by UEA1 modified liposomes were (84 +/- 15)% at 4 h, (78 +/- 11)% at 8 h and (90 +/- 12)% at 12 h after oral administration. The conventional liposomes and saline showed no effect. The blood glucose levels of the diabetic mice reduced by UEA1 modified liposomes were (73 +/- 7)% at 4 h, (74 +/- 9)% at 8 h, (86 +/- 9)% at 12 h after oral administration. CONCLUSION: The UEA1 modified liposomes promote the oral absorption of insulin due to the specific-site combination on M cell membrane. PMID- 15813032 TI - Interaction of a novel peptoid enhancer--arginine oligomer with bovine submaxillary mucin. AB - AIM: To determine the thermodynamics of binding reaction of arginine oligomer (R8) to bovine submaxillary mucin (BSM) in order to provide the foundation for understanding the influence of mucin on transport of macromolecules through mucosa mediated by arginine oligomer. METHODS: Ultracentrifugation sedimentation was employed to investigate the interaction of BSM-R8. The mixtures of R8 with variable concentration and constant volume of BSM were placed on a shaker under oscillation at 25 degrees C to achieve equilibriums of binding reaction, and then centrifuged. The fluorescence intensity of the supernatant was measured by spectrofluorometer. The data were described by two types of binding sites model, the binding parameters of BSM-R8 were obtained by Scatchard plots. RESULTS: At the low pH values < or = 4.5 and ionic strength > or = 0.2 mol x L(-1), the BSM R8 interaction was principally electrostatic interaction, the five primary binding sites (n1) predominantly were supplied by sulfate groups, the secondary binding sites apparently depended on pH, in that percent ionization of sialic acid residues (n2) in BSM. At the low ionic strength < or = 0.2 mol x L(-1) and pH 7.0, the BSM-R8 interaction was exceedingly complex, hydrogen bonds, hydrophobic interaction and electrostatic forces were involved in the interaction between R8 and BSM, the binding sites of BSM bound R8 were markedly increased. CONCLUSION: There existed evidence that R8 interacted with BSM. The pH and the ionic strength of the binding solution strongly affected the interaction of BSM with R8. The results suggested that the enhancing efficacy of the arginine oligomer for the transport of macromolecules through different site mucosa in body might be variable. PMID- 15813033 TI - [Influence of different doxorubicin-salt aggregates on the leakage of liposomal doxorubicin in vitro and long circulation in vivo]. AB - AIM: To develop liposomes containing doxorubicin with different salts and to investigate their influence on the stability of liposomal doxorubicin in vitro and in vivo. METHODS: Liposomes were prepared by the film method, treated further by extruded through nuclear membrane. The entrapment efficiency was determined by column chromatography. In vitro drug release experiments were carried out with a dialysis bag (Mw cut-off 12000 - 14000). Reverse-phase HPLC was used to study the pharmacokinetics of liposomal doxorubicin. RESULTS: The particle size of liposomes with glycinate buffer, citrate buffer and ammonium sulfate as the inner water phase were (103 +/- 8), (102 +/- 12) and (97 +/- 8) nm. The zeta potential and the encapsulation ratio were (-21.3 +/- 0.5), (-21.7 +/- 0.4), (-20.9 +/- 0.7) mV and 47.8%, 96.7%, 98.6%, respectively. The leaking rate of doxorubicin from liposomes was related to the pH value of the release medium. The leaking rate increased at lowered pH. Pharmacokinetic study showed that the MRT (mean retention time) of liposomes with glycinate buffer, citrate buffer and ammonium sulfate as the inner water phase were 12.13, 23.31 and 29.79 h, respectively. CONCLUSION: Doxorubicin showed different stability in liposomes with different inner water phases, the weaker the acid in the inner water phase, the stabler the liposome. PMID- 15813034 TI - [Preparation of cyclosporine A pH sensitive nanoparticles and oral pharmacokinetics in rats]. AB - AIM: To study the preparation conditions and its oral pharmacokinetic characteristics of cyclosporine A (CyA) pH sensitive nanoparticles. METHODS: The CyA pH sensitive nanoparticles were prepared by the quasi-emulsion solvent diffusion technique (QESD). Male Sprague-Dawley (SD) rats weighing (250 +/- 20) g were selected and randomly divided into five groups. The bioavailability of CyA from nanoparticles and Neoral microemulsion were assessed at a dose of 15 mg x kg(-1) by gavage. The concentration of CyA in whole blood samples was detected by HPLC to evaluate the relative bioavailability of CyA pH sensitive nanoparticles. RESULTS: The blood concentration profiles of CyA pH sensitive nanoparticles in rats fitted to two compartment models using 3P87 pharmacokinetic calculation program. Compared with the Neoral microemulsion, the relative bioavailability of CyA was 94.8%, 115.2%, 113.6% and 132.5% for CyA-E100, CyA-L100, CyA-L100-55 and CyA-S100 nanoparticles respectively. CONCLUSION: CyA-S100 nanoparticles was shown to significantly improve the oral bioavailability of CyA compared with Neoral microemulsion (P < 0.05). While there were no significant differences between Neoral microemulsion and other CyA pH sensitive nanoparticles. With these results, the potential of pH-sensitive nanoparticles for the oral delivery of CyA was confirmed. Furthermore, this formulation approach can be used to improve the oral bioavailability of other poorly soluble and poorly absorbable drugs. PMID- 15813035 TI - [Recent advances in the study on poly- and oligo-saccharides with hypoglycemic activity]. PMID- 15813036 TI - [History and concepts of the renin-angiotensin system]. PMID- 15813037 TI - [A new paradigm for renin research]. PMID- 15813038 TI - [Gene, biosynthesis, and function of angiotensinogen]. PMID- 15813039 TI - [ACE: gene, biosynthesis and functions]. PMID- 15813040 TI - [Angiotensin receptor subtype and signal transduction]. PMID- 15813041 TI - [Tissue distribution and regulatory mechanism of angiotensin II receptor expression]. PMID- 15813042 TI - [Genetic dissection of the renin-angiotensin system]. PMID- 15813043 TI - [Genetically engineered animals for angiotensin II receptor]. PMID- 15813044 TI - [Measurement and evaluation of renin-angiotensin system]. PMID- 15813045 TI - [Chymase-dependent angiotensin II-formation]. PMID- 15813046 TI - [Role of rennin-angiotensin system in cardiac remodeling]. PMID- 15813047 TI - [Renin-angiotensin system and vascular function and remodeling]. PMID- 15813048 TI - [Regulation of renal functions by renin-angiotensin system]. PMID- 15813049 TI - [Renin-angiotensin system and apoptosis]. PMID- 15813050 TI - [The renin-angiotensin system and insulin resistance]. PMID- 15813051 TI - [Renin-angiotensin system and oxidative stress]. PMID- 15813052 TI - [Renin-angiotensin system and angiogenesis]. PMID- 15813053 TI - [Effects of renin-angiotensin system in the central nervous system]. PMID- 15813054 TI - [Essential hypertension]. PMID- 15813055 TI - [Renovascular hypertension]. PMID- 15813056 TI - [Salt-sensitive hypertension]. PMID- 15813057 TI - [Reninoma]. PMID- 15813058 TI - [Malignant hypertension]. PMID- 15813059 TI - [Renin-angiotensin system in congestive heart failure]. PMID- 15813060 TI - [Renal failure--nephrosclerosis]. PMID- 15813061 TI - [Diabetic nephropathy]. PMID- 15813062 TI - [Atherosclerosis]. PMID- 15813063 TI - [Hypokalemic nephropathy]. PMID- 15813064 TI - [Bartter' s syndrome]. PMID- 15813065 TI - [Renin-angiotensin system and metabolic domino]. PMID- 15813066 TI - [Renin-angiotensin system blockers and organ protection]. PMID- 15813067 TI - [Combination therapy of angiotensin converting enzyme inhibitors and angiotensin II receptor blockers]. PMID- 15813068 TI - [Blockade of renin-angiotensin system and aldosterone breakthrough]. PMID- 15813069 TI - [Discovery and significance of aldosterone]. PMID- 15813070 TI - [Aldosterone biosynthesis and related enzyme genes]. PMID- 15813071 TI - [Mechanism of the regulation of aldosterone secretion]. PMID- 15813072 TI - [Mineralocorticoid receptor and its signal transduction]. PMID- 15813073 TI - [Distribution of mineralocorticoid receptor in human]. PMID- 15813074 TI - [Genetic manipulation animals of aldosterone-related genes]. PMID- 15813075 TI - [Measurement and evaluation of aldosterone]. PMID- 15813076 TI - [Renal action of aldosterone]. PMID- 15813077 TI - [Cardiac aldosterone production]. PMID- 15813078 TI - [Vascular action of aldosterone]. PMID- 15813079 TI - [Role of aldosterone in central nervous system]. PMID- 15813080 TI - [Clinical entity of primary aldosteronism including idiopathic hyperaldosteronism and glucocorticoid remediable aldosteronism]. PMID- 15813081 TI - [Pathology of primary aldosteronism]. PMID- 15813082 TI - [11Beta-hydroxylase deficiency]. PMID- 15813083 TI - [17Alpha-hydroxylase deficiency]. PMID- 15813084 TI - [Deficiency of 11beta-hydroxysteroid dehydrogenase type 1]. PMID- 15813085 TI - [Syndrome of apparent mineralocorticoid excess]. PMID- 15813086 TI - [Gordon's syndrome and Gitelman's syndrome]. PMID- 15813087 TI - [Liddle syndrome]. PMID- 15813088 TI - [Aldosterone and heart failure]. PMID- 15813089 TI - [Left ventricular remodeling post myocardial infarction]. PMID- 15813090 TI - [Randomized Aldactone Evaluation Study (RALES)]. PMID- 15813091 TI - [Eplerenone Post-AMI Heart Failure Efficacy and survival study (EPHESUS)]. PMID- 15813092 TI - [Comparative studies on prevalence of primary aldosteronism and on diagnostic procedures]. PMID- 15813093 TI - [Discovery and significant of kallikrein-kinin system]. PMID- 15813094 TI - [Gene structure, biosynthesis and function of kininogen]. PMID- 15813095 TI - [The gene, synthesis, function of kallikrein]. PMID- 15813096 TI - [Structure and signaling pathways of kinin receptors]. PMID- 15813097 TI - [Distribution and regulation of kallikrein-kinin system]. PMID- 15813098 TI - [Transgenic and knockout models for the kallikrein-kinin system]. PMID- 15813099 TI - [Measurements and pathophysiological assessments for kallikrein-kinin system]. PMID- 15813100 TI - [Significance of kallikrein-kinin system on blood pressure regulation]. PMID- 15813101 TI - [Kallikrein-kinin system and its regulation for water and electrolyte metabolism]. PMID- 15813102 TI - [Role of kallikrein-kinin system in glucose metabolism]. PMID- 15813103 TI - [Eessential hypertension]. PMID- 15813104 TI - [Secondary hypertension]. PMID- 15813105 TI - [Ischemic heart disease]. PMID- 15813106 TI - [Kinin antagonist]. PMID- 15813107 TI - [Multiple interactions between the kallikrein-kinin and the renin angiotensin systems]. PMID- 15813108 TI - [Discovery of vasopressin and its significance]. PMID- 15813109 TI - [Gene, secretion and function of arginine vasopressin]. PMID- 15813110 TI - [Vasopressin receptor and signal transduction]. PMID- 15813111 TI - [Regulation of gene expression and release of vasopressin]. PMID- 15813112 TI - [Transgenic, knock-out and knock-in animal models for vasopressin neuron systems]. PMID- 15813113 TI - [Measuring methods of vasopressin and its evaluation]. PMID- 15813114 TI - [Function of vasopressin in kidney]. PMID- 15813115 TI - [Vascular effects of vasopressin]. PMID- 15813116 TI - [Effects of central arginine vasopressin on the sympathetic nervous system]. PMID- 15813117 TI - [Augmented baroreflex sensitivity by vasopressin]. PMID- 15813118 TI - [Diabetes insipidus]. PMID- 15813119 TI - [Syndrome of inappropriate secretion of ADH (SIADH)]. PMID- 15813120 TI - [Essential hypertension]. PMID- 15813121 TI - [Chronic heart failure]. PMID- 15813122 TI - [Genetic disorders of vasopressin and vasopressin receptor genes]. PMID- 15813123 TI - [Perspectives of vasopressin receptor antagonists]. PMID- 15813124 TI - [The discovery of catecholamine and its clinical significance]. PMID- 15813125 TI - [Pheochromocytoma]. PMID- 15813126 TI - [Abnormality of sympathetic function]. PMID- 15813127 TI - [Roles of catecholamine in essential hypertension]. PMID- 15813128 TI - [The role of catecholamine in the onset and progression of heart failure]. PMID- 15813129 TI - [Cardiac hypertrophy]. PMID- 15813130 TI - [Perspective of sympathetic nervous system inhibitors]. PMID- 15813131 TI - [The role of estrogen in the regulation of blood vessel function]. PMID- 15813132 TI - [Estrogen--hypertension in menopause]. PMID- 15813133 TI - [Myocardial infarction]. PMID- 15813134 TI - [Cerebral infarction]. PMID- 15813135 TI - [Arteriosclerosis and estrogen]. PMID- 15813136 TI - [Hormone replacement therapy and prevention of cardiovascular diseases]. PMID- 15813137 TI - [Selective estrogen receptor modulator (SERM)]. PMID- 15813138 TI - [Crosstalk between angiotensin II receptor subtypes and estrogen]. PMID- 15813139 TI - Search and rescue is search and rescue--or, the basics still apply. PMID- 15813140 TI - The effects of 10 weeks of reforestation work on body composition. AB - OBJECTIVE: To document changes in body composition and body mass in male and female tree planters. METHODS: Height, mass, skin-fold thickness, and limb girths were measured in 17 male and 5 female tree planters before and after 10 weeks of work. RESULTS: Significant decreases were found in body mass (80.6 +/- 10.7 kg vs 76.8 +/- 8.5 kg) and body fat (13.3% +/- 5.5% vs. 10.4% +/- 5.0%) in the men (P < .05). No changes in skin-fold-corrected limb girths were found in the men or women. Initial body mass was significantly (P < .05) correlated with mass loss in men (r2 = .46) and women (r2 = .67). Estimated daily energy consumption was 20680.1 +/- 2204.5 kJ for men and 14516.6 +/- 2077.3 kJ for women, and estimated daily fat consumption was 194.2 +/- 30.1 g for men and 132.3 +/- 35.6 g for women. CONCLUSIONS: Ten weeks of tree planting leads to significant decreases in body mass and body fat in men while maintaining skin-fold-corrected limb girths. PMID- 15813141 TI - Passive prophylaxis with permethrin-treated tents reduces mosquito bites among North American summer campers. AB - OBJECTIVE: Over 2 million adolescents participate in summer-camp experiences, placing themselves at risk for mosquito-borne illness. Insect repellent is recommended but often not used. It is unknown whether permethrin treatment of a location, such as a campsite, provides passive prophylaxis reducing mosquito bites among all persons in the immediate vicinity. METHODS: This randomized, double-blind controlled trial used 0.4% permethrin (2.5% permethrin diluted by 7 parts water) sprayed once onto the external surface of canvas tents at a Boy Scout summer camp. During a 9-week period, subjects (n = 545) completed nightly mosquito-bite and -landing counts for 5 minutes at dusk and recorded insect repellent use within 2 hours of counting. Weekly mosquito sampling with CO2 baited Centers for Disease Control and Prevention traps occurred at study campsites. The primary outcome measure was the number of mosquito bites per 5 minutes. RESULTS: A total of 1614 person nights averaged 5.1 +/- 7.2 (+/- SD) mosquito bites per 5 minutes. The permethrin-treated campsites had decreased mosquito landings and bites (relative risk reduction [RRR] 44%; 95% CI 34% to 55%; P < .001) compared with controls. Insect repellent was used only 32% of nights, decreasing bites by 36% (RRR 36%; 95% CI 25% to 47%; P < .001). Permethrin was superior to insect repellent alone (RRR 20%; 95% CI 4% to 37%; P = .01). CONCLUSIONS: Permethrin treatment of tents is an effective, inexpensive public health measure to reduce mosquito bites. Permethrin is effective among all individuals in a camping setting and was more effective than topical insect repellent alone, which, although recommended, was inconsistently used. PMID- 15813142 TI - Physicians and lay people are unable to apply pressure immobilization properly in a simulated snakebite scenario. AB - OBJECTIVE: To determine whether volunteers (with or without prior medical training) can correctly apply pressure immobilization (PI) in a simulated snakebite scenario after receiving standard instructions describing the technique. METHODS: Twenty emergency medicine physicians (residents and attendings) and 20 lay volunteers without prior formal medical training were given standard printed instructions describing the application of PI for field management of snakebite. They were then supplied with appropriate materials and asked to apply the technique five separate times (twice to another individual [one upper and one lower extremity] and three times to themselves [nondominant upper extremity, dominant upper extremity, and one lower extremity]). Successful application was defined a priori by four criteria previously published in the literature: wrap begins at the bite site, entire extremity is wrapped, splint or sling is applied, and pressures under the dressing are between 40 and 70 mm Hg in upper-extremity application and between 55 and 70 mm Hg in lower-extremity use. Pressures were determined using a specially designed skin interface pressure measuring device placed at the simulated bite site. RESULTS: The technique was correctly applied as judged by the preset criteria in only 13 out of 100 applications by emergency medicine physicians and in only 5 out of 100 applications by lay people. There was no significant difference in success rates between physicians and lay volunteers. Likewise, there was no significant difference in success based on which extremity was being wrapped. More detailed analysis revealed that the major contributor to failure was inability to achieve recommended target pressures. CONCLUSIONS: Volunteers in a simulated snakebite scenario have difficulty applying PI correctly, as defined in the literature. The major source of failure is an inability to achieve recommended pressure levels under the dressing. New methods of instructing people in the proper use of PI or new technologies to guide or automate application are needed if this technique is to be used consistently in an effective manner for field management of bites by venomous snakes not known to cause significant local wound necrosis. PMID- 15813143 TI - Effect of hypoxia on norepinephrine of various tissues in rats. AB - OBJECTIVE: To determine the effects of hypoxia and hypoxic exercise (HE) on the norepinephrine levels of various tissues in rats. METHODS: Male Wistar rats were randomly assigned to 3 groups: an HE group (n = 6), a hypoxic-sedentary (HS) group (n = 6), and a normoxic-sedentary (NS) group (n = 6). The HE rats had access, ad lib, to an exercise wheel for 8 weeks. HE and HS rats were maintained in a normobaric hypoxic chamber with an FIO2 of 16%. Norepinephrine levels were measured and compared in liver, heart, diaphragm, soleus, and gastrocnemius tissues from the 3 groups. RESULTS: Liver norepinephrine levels in the HE and HS groups were significantly lower than the levels in the NS group (P < .05). No significant difference was found in liver norepinephrine levels between the HE and the HS groups. The heart norepinephrine levels in the NS group were significantly lower than the levels in the HE (P < .01) and HS groups (P < .01). In contrast, no significant differences were found in the norepinephrine levels for the diaphragm and soleus muscle among the 3 groups. The norepinephrine levels in the gastrocnemius white muscles were significantly higher in the HS group than in the HE (P < .05) and NS groups (P < .01). P < .01 represents a significant difference at the level of 1%. CONCLUSIONS: This study demonstrated that hypoxia and HE both elicit a decreased sympathetic response in the liver tissue of male Wistar rats but cause an increased response in heart tissue. These results suggest that the sympathetic responses to long-term hypoxia and HE training are different in various rat tissues. PMID- 15813144 TI - Surviving atmospheric spacecraft breakup. AB - Spacecraft travel higher and faster than aircraft, making breakup potentially less survivable. As with aircraft breakup, the dissipation of lethal forces via spacecraft breakup around an organism is likely to greatly increase the odds of survival. By employing a knowledge of space and aviation physiology, comparative physiology, and search-and-rescue techniques, we were able to correctly predict and execute the recovery of live animals following the breakup of the space shuttle Columbia. In this study, we make what is, to our knowledge, the first report of an animal, Caenorhabditis elegans, surviving the atmospheric breakup of the spacecraft that was supporting it and discuss both the lethal events these animals had to escape and the implications for search and rescue following spacecraft breakup. PMID- 15813145 TI - Spectrum of injuries from glacial sports. AB - OBJECTIVE: Glacial sports continue to be a popular form of wilderness activity, but no published studies have commented on this type of sport or focused on the injury patterns of glacial accidents. The objectives of this study were to investigate the types and frequency of injuries associated with a glacial crevasse or ice-field fall. METHODS: The trauma registry data and radiology record system of a pediatric and adult level 1 trauma center were evaluated from January 1997 through August 2003. All admissions with injuries caused by a glacial-crevasse or ice-field fall were reviewed. RESULTS: During the 5 1/2-year period we identified 12 patients, of whom 9 (75%) were male. The average age was 30.2 years with an age range of 9 to 57 years. Six patients fell into glacial crevasses, and the other 6 slid down an ice field of a glacier. Leading diagnoses were head injury (44.6%), thorax injury (20.1%), and lower extremity injury (18.4%). Serious injuries were observed in 66.6% of the patients with a Glasgow Coma Scale of 3 to 8. CONCLUSIONS: A wide spectrum of injuries is associated with glacial accidents. They range from common extremity injuries with fracture or joint instability and peripheral frostbite to potentially life-threatening nonorthopedic trauma, which requires intensive clinical and radiological work-up. Injury prevention strategies should focus on wearing helmets to reduce head injuries and adequate clothing to forestall or prevent hypothermia, for these are the most severe and life-threatening injuries. PMID- 15813146 TI - Change in differential diagnosis and patient management with the use of portable ultrasound in a remote setting. AB - OBJECTIVE: Physicians practicing in remote areas are typically limited in their choice of diagnostic tools. The goal of this study was to determine whether the use of a portable ultrasound (US) device on selected patients in a remote setting would alter physician diagnosis and management. METHODS: This was a prospective observational study of the affects of US on physician decision making deep in the Amazon jungle. A battery-operated Sonosite 180 Plus with 2 interchangeable transducers (4-7-MHz broadband intercavitary transducer and 2-5-MHz broadband abdominal transducer) was used. The patient population consisted of local tribal people. Two of the physicians on the team performed all US examinations. Team physicians requesting US examinations filled out a survey before and after the US examination. Before the US, the referring physician filled out a survey describing the patient's initial complaint, pertinent past medical history and physical findings, and an initial (pre-US) differential diagnosis and planned treatment with expected disposition. After the results of the US were reviewed with the referring physicians, the doctors were asked to fill out the remainder of the survey, allowing comparison of pre- and post-US differential diagnosis, treatment plan, and disposition. RESULTS: A total of 25 US studies were performed during this study (1 trauma US scan, 6 hepatobiliary studies, 5 transabdominal pelvic scans, 7 transvaginal pelvic studies, 3 renal studies, and 3 abdominal aortic scans). The monitor on the US unit experienced a rare failure shortly after being used at 17,000 ft and then 10 times at sea level, and no further US scans could be performed. US scan results dramatically altered the disposition of 7 patients, including 4 patients who avoided a potentially dangerous 2-day evacuation to more definitive medical care. Three patients were found to need rapid referral to the nearest clinic for surgical evaluation. CONCLUSIONS: When used in a remote location, portable US provides a significant benefit that can dramatically alter disposition and treatment. PMID- 15813147 TI - Ataxia at altitude measured on a wobble board. AB - OBJECTIVE: To establish a simple measure of ataxia for use at high altitude. METHODS: Twenty healthy subjects took part in a trek to 5005 m. At 5 different altitudes on the route, they undertook a balance test using a wobble board. The primary objectives were to investigate disturbances of ataxia at altitude and to correlate any observed disturbances with acute mountain sickness (AMS) scores. Secondary outcomes were correlations with cerebral regional oxygenation, pulse oximetry, and age. RESULTS: After a short learning curve, the wobble board test was found to be reproducible. Subjects over 31 years of age were significantly less steady than younger subjects. Subjects suffering acute mountain sickness scored significantly worse on the wobble board test, although scores did not correlate with a specific question on unsteadiness. A positive test defined as equal to or more than 2.5 contacts over 2 minutes gave a predictive value for acute mountain sickness of 66.7% at 4650 m and 100% at 5005 m. Cerebral regional oxygenation in 9 subjects at 5005 m correlated with the wobble board test (r = 0.73; p < .05), whereas pulse oximetry did not. CONCLUSIONS: The wobble board may be a useful adjunct in quantitating ataxia in the field. A positive result may indicate the presence of AMS and may be a useful clinical measure of cerebral hypoxia but should be correlated with other clinical features. PMID- 15813148 TI - Tibetans at extreme altitude. AB - Between 1960 and 2003, 13 Chinese expeditions successfully reached the summit of Chomolungma (Mt Everest or Sagarmatha). Forty-five of the 80 summiteers were Tibetan highlanders. During these and other high-altitude expeditions in Tibet, a series of medical and physiological investigations were carried out on the Tibetan mountaineers. The results suggest that these individuals are better adapted to high altitude and that, at altitude, they have a greater physical capacity than Han (ethnic Chinese) lowland newcomers. They have higher maximal oxygen uptake, greater ventilation, more brisk hypoxic ventilatory responses, larger lung volumes, greater diffusing capacities, and a better quality of sleep. Tibetans also have a lower incidence of acute mountain sickness and less body weight loss. These differences appear to represent genetic adaptations and are obviously significant for humans at extreme altitude. This paper reviews what is known about the physiologic responses of Tibetans at extreme altitudes. PMID- 15813149 TI - Adolescents at altitude. PMID- 15813150 TI - In-hospital death in patients with chronic obstructive pulmonary disease. PMID- 15813151 TI - Sarcoidosis presenting as acute respiratory failure. PMID- 15813152 TI - Peak expiratory flow rate as predictor of inpatient death in patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: Few studies analyze hospital deaths and related factors in patients with acute exacerbation of chronic obstructive pulmonary disease who require hospitalization. METHODS: A cross-sectional study was done with 284 patients who had been admitted consecutively to the Short Stay Medical Unit at the Juan Canalejo Hospital in A Coruna. RESULTS: Eleven patients (3.9%) died. The independent variables for predicting death were the peak expiratory flow (OR, 0.96; 95% CI, 0.94 to 0.98), long-term oxygen therapy (OR, 12.46; 95% CI, 2.1 to 72.4), and body mass index (OR, 0.73; 95% CI, 0.59 to 0.90). A peak expiratory flow < 150 L/min showed the best specificity and positive predictive value with maximum sensitivity for predicting death. The results of the arterial blood gasses and the functional tests did not predict hospital death. CONCLUSIONS: Peak expiratory flow was the most important predictive value for determining the risk of death in patients who required hospitalization for acute exacerbation of chronic obstructive pulmonary disease. Additional studies are required to validate these findings. PMID- 15813153 TI - Enteral nutrition by a forward surgical team in Afghanistan. AB - OBJECTIVES: The modern practice of trauma surgery is a global physiologic approach to caring for the injured patient. Included in that approach is consideration of the traumatized patient's nutritional needs and implementing early enteral feeding. This is routine practice in the United States but logistically impractical when using commercial enteral feeding formulas in the austere environment of a Forward Operating Base in Afghanistan. METHODS: At a Forward Operating Base in southern Afghanistan, injured patients who were not expected to be taking a regular diet by 72 hours after injury are started on early enteral feedings. This is through nasogastric, gastrostomy, or jejunostomy tube, using formulas of pureed food available in the theater from local sources and supplemented with ingredients from US Army Meals-Ready-to-Eat. Preparation, nutritional calculation, and delivery are discussed. RESULTS: The injured soldiers and Afghan nationals tolerated early enteral feedings with no complications encountered related to the feeding portal, infusion, or ingredients. CONCLUSIONS: Early enteral feeding of traumatized patients in the far forward environment of southeastern Afghanistan is practical with the use of ingredients found locally in-theater. PMID- 15813154 TI - Music as therapy. AB - Music is widely used to enhance well-being, reduce stress, and distract patients from unpleasant symptoms. Although there are wide variations in individual preferences, music appears to exert direct physiologic effects through the autonomic nervous system. It also has indirect effects by modifying caregiver behavior. Music effectively reduces anxiety and improves mood for medical and surgical patients, for patients in intensive care units and patients undergoing procedures, and for children as well as adults. Music is a low-cost intervention that often reduces surgical, procedural, acute, and chronic pain. Music also improves the quality of life for patients receiving palliative care, enhancing a sense of comfort and relaxation. Providing music to caregivers may be a cost effective and enjoyable strategy to improve empathy, compassion, and relationship centered care while not increasing errors or interfering with technical aspects of care. PMID- 15813155 TI - Approach to communicating with patients about the use of nutritional supplements in cancer care. AB - In recent years, complementary and alternative medicine has become popular among the general population in the Western world. Cancer patients have joined this global trend, often seeking supplements to conventional oncologic care, usually without their physicians' knowledge. Among the most common forms of complementary and alternative medicine used by cancer patients are natural products such as herbs and megavitamins. The extensive use of nutritional supplements by cancer patients raises multiple questions and challenges for the physician. Since there are limited scientific data on the efficacy and safety of many nutritional supplements, advising patients about when to use them during the course of illness is difficult. This is true for each stage of cancer care: prevention, acute active care (radiation, chemotherapy, surgery), and post-acute care (follow up visits and prevention of recurrence). The authors describe a patient-centered approach to the use of nutritional supplements in cancer care. PMID- 15813156 TI - Pain management in children: developmental considerations and mind-body therapies. AB - One of the most challenging roles of medical providers serving children is to appropriately assess and treat their pain. Pain is one of the most misunderstood, underdiagnosed, and undertreated/ untreated medical problems, particularly in children. New JCAHO regulations regard pain as "the fifth vital sign" and require caregivers to regularly assess and address pain. This review focuses on the clinical assessment of pain, based on a developmental model and addresses common beliefs and myths that affect the management of pain in children. We provide a review of the pain literature that focuses on the integration of mind-body therapies into the management of procedure-related pain, headache, and recurrent abdominal pain in children. PMID- 15813157 TI - Inflammation: nutritional, botanical, and mind-body influences. AB - Chronic inflammation is becoming an important risk factor to identify in regard to inhibiting disease onset and its progression. Nutritional science attempted to improve health by manipulating fats so that we could consume "healthy" nonsaturated fats while simultaneously allowing foods to have a longer shelf life. However, despite our good intentions, trans-fats and partially hydrogenated oils have been found to promote inflammation and adversely affect health. This article reviews how essential fatty acids, the ratio of omega-3 to omega-6 fatty acids, glycemic load, the Mediterranean diet, specific foods and botanicals, and the mind-body relation influence the inflammatory cascade. PMID- 15813158 TI - Use of Echinacea in upper respiratory tract infection. AB - The significant burden of upper respiratory tract infection in adults and children, coupled with a lack of specific treatment options, invites the use of alternative therapies. Echinacea is an herb widely used for the prevention or treatment of upper respiratory tract infection. This review article examines the mechanism of action, dose, and types of Echinacea used for these purposes. The principal mode of action of Echinacea is through immunostimulation. Most Echinacea studies were done in Germany, but their results are difficult to interpret because of variability of experimental parameters. Types of Echinacea commonly used are Echinacea purpurea, E pallida, and E angustifolia. Both the plant's upper parts and roots are used. For oral administration, tablets, extracts, fresh pressed juice, teas, and tinctures have been used. Though studies show a beneficial effect, clear conclusions and recommendations of Echinacea use cannot be made due to a lack of standard product, variability in dose, and variability in outcome measures. Therefore, well-designed studies with consistent standardized measures are required. PMID- 15813159 TI - Integrative therapies for menopause. AB - Menopause is a transitional time for women. This gives practitioners an opportunity to focus on recommending healthy life-style changes. Hormone replacement therapy (HRT) has been the mainstay of therapy for menopausal symptoms. With recent research findings, women and their physicians are seeking alternatives that do not carry the risks associated with HRT. Exercise has been shown to help some women with symptoms of hot flashes, as have relaxation techniques and deep breathing. Dietary changes to incorporate whole foods and soy are thought by some to help with menopausal symptoms, and are recommended because of a positive impact on heart disease and obesity; soy isoflavones may also help with menopausal symptoms. Botanicals such as black cohosh and red clover have been shown in some studies to decrease severity and frequency of hot flashes. We recommend that HRT be prescribed when other measures have failed to adequately control symptoms. Bioidentical hormones are preferred in our practice. PMID- 15813160 TI - Acupuncture: a clinical review. AB - This article summarizes the research base, probable mechanism of actions, and clinical applications of acupuncture. It offers the clinician a deeper understanding of appropriate conditions for which acupuncture may be useful, outlines how to integrate acupuncture into a clinical practice, and describes referral and training issues. PMID- 15813161 TI - Neoadjuvant therapy: an emerging concept in oncology. AB - Neoadjuvant therapy, an adjunctive therapy given before the main therapy, has become an integral part of modem multidisciplinary cancer management. Organized by the primary organ involved by cancer, this review summarizes the outcomes of neoadjuvant therapy for common malignant solid tumors, based on large, randomized, controlled trials. In locally advanced rectal, laryngeal, and breast cancer, neoadjuvant therapy enables organ preservation; however, it does not improve overall survival when compared with definitive treatment followed by adjuvant therapy. In locally advanced bladder and cervical cancer, patients who undergo neoadjuvant therapy before radical surgery appear to have better survival than those receiving definitive therapy alone; however, it is unclear if the neoadjuvant approach will be superior to definitive therapy followed by adjuvant therapy. To date, the survival benefits of neoadjuvant therapy for resectable non small cell lung, esophageal, gastric, and prostate cancer remains under investigation. PMID- 15813162 TI - Extrapulmonary small cell carcinoma. AB - This article reviews the recent literature on extrapulmonary small cell carcinomas. Until now, only four cases have been published in the English literature, two of those in the Southern Medical Journal. Sharing the information on diagnosis and treatment of these cases is important for better understanding and treatment of these patients. The differential diagnosis of pleural masses and the relation to asbestos exposure is briefly discussed. In addition, the available literature on other extrapulmonary localizations of small cell carcinoma is reviewed. The focus is on the available larger series of patients. Small cell carcinoma of the skin is included. This form of small cell carcinoma is often excluded from the larger review series, but can be an important alternative in the diagnostic process of patient evaluation. PMID- 15813163 TI - Substance abuse: medical and slang terminology. AB - Substance abuse is among one of the major problems plaguing our society. It has come to the attention of several healthcare professionals that a communication gap exists between themselves and substance abusers. Most of the time the substance abusers are only familiar with the slang terms of abused substances, a terminology that medical professionals are usually unaware of. This paper is an attempt to close that communication gap, allowing health care professionals to understand the slang terminology that their patients use, thus enabling them to make appropriate treatment decisions. In addition, the article presents some key features (including active ingredient, pharmacological classification, medical use, abuse form, usage method, combinations used, effects sought, long-term possible effects, and detectability in urine) of the most commonly abused substances. PMID- 15813164 TI - Strategies for insulin therapy in type 2 diabetes. AB - Chronic hyperglycemia in type 2 diabetes is responsible for an array of microvascular and macrovascular complications that can lead to significant morbidity and mortality. Several well-conducted large clinical studies have shown that normalizing blood glucose levels can help prevent the onset and slow the progression of complications from diabetes. As many as 25% of patients treated with oral hypoglycemic agents require the addition of insulin therapy to compensate for the progression of beta-cell failure and an inability to maintain glycemic control. Various strategies incorporating the use of insulin early in the course of the disease have been developed to meet this goal, and include the use of basal-bolus insulin regimens as well as bedtime insulin injections. The pharmacokinetic properties of the new insulin analogs (eg, insulin lispro, insulin aspart, insulin glargine) offer significant advantages, such as improved control of nocturnal hypoglycemia with basal insulin glargine, and improved postprandial glucose control, with insulin lispro or insulin aspart. PMID- 15813165 TI - Critical care aspects of alcohol abuse. AB - The authors reviewed MEDLINE and references of major articles in the published literature over the last 30 years regarding the complications of alcohol abuse and discuss the critical care aspects of alcohol abuse. This article discusses the severe medical conditions associated with alcohol abuse that lead to admission to the medical intensive care unit. The clinical manifestations, pathophysiology, diagnostic studies, and management of these conditions are discussed in detail. PMID- 15813166 TI - Pulmonary sarcoidosis presenting with acute respiratory failure. AB - This is a case report of pulmonary sarcoidosis presenting with acute respiratory failure. A 33-year-old black man presented with 4 days of headache and fever. He had subsequent development of respiratory failure on the seventh hospital day. Workup showed high serum angiotensin-converting enzyme and increased pulmonary parenchymal uptake of gallium. Extensive workup for collagen vascular disease and infectious pathology were negative. Transbronchial biopsy specimens of the lung parenchyma showed noncaseating epithelioid granulomas consistent with sarcoidosis. The patient was treated with intravenous methylprednisolone succinate with dramatic clinical improvement. PMID- 15813167 TI - Acupuncture therapy rapidly terminates intractable hiccups complicating acute myocardial infarction. AB - Acupuncture is a well-known alternative therapy in practice worldwide. Its dramatic effect on hiccups has been rarely reported. We describe a 77-year-old male who had hiccups after an acute myocardial infarction. Despite aggressive treatment including breath-holding to interrupt the respiratory rhythm, continuous positive airway pressure, and medication with metoclopramine, prochlorperazine, chlorpromazine, haloperidol, mephenesin, diphenylhydantoin, baclofen, and phenobarbital, the hiccups persisted for 7 days. Eventually, the hiccups were rapidly terminated by acupuncture at acupoint GV14 (Da zhui). To the best of our knowledge, this is the first report of acupuncture's reversing intractable hiccups after an acute myocardial infarction. Acupuncture may be considered for patients with hiccups refractory to conventional therapy. PMID- 15813168 TI - Primary non-hodgkin lymphoma of the larynx. AB - A rare case of primary large B-cell non-Hodgkin lymphoma of the larynx is reported. The patient was an 80-year-old female who presented with pharyngeal discomfort and dysphagia. Radiotherapy was instituted with complete remission of the tumor. Nine months afterward, she presented with aspiration pneumonia and dysphagia. Indirect laryngoscopy disclosed laryngeal and esophageal edema, which was probably related to radiotherapy. No signs of tumor recurrence were observed. Three months later, she was entirely asymptomatic. Although cases of aggressive course and poor response to therapy exist, primary large B-cell non-Hodgkin lymphoma of the larynx is generally considered a relatively benign and radiosensitive tumor. PMID- 15813169 TI - Acute Epstein-Barr virus infection causing bilateral conjunctival hemorrhages. AB - The systemic and ocular manifestations of acute Epstein-Barr virus infection are protean. Conjunctival hemorrhage has been described once. This report describes a young male who had bilateral conjunctival hemorrhages in the setting of acute Epstein-Barr virus infection. PMID- 15813170 TI - Thrombotic thrombocytopenic purpura without schistocytes on the peripheral blood smear. AB - A hallmark of the clinical syndrome of thrombotic thrombocytopenic purpura (TTP) is evidence of microangiopathic hemolytic anemia. The presence of schistocytes on the peripheral blood smear, elevated plasma lactic dehydrogenase, and decreased haptoglobin concentration are used as evidence of microangiopathic hemolytic anemia to make a diagnosis of TTP. This report describes a case of recurrence of TTP in the absence of schistocytes in the peripheral blood smear during the recurrent episode. Although careful attention should be paid to microscopic examination of a blood smear in any patient presenting with acute renal failure and thrombocytopenia, this case emphasizes the need to consider TTP-hemolytic uremic syndrome in the differential diagnosis, even in the absence of peripheral schistocytosis. PMID- 15813171 TI - On omeprazole and iron replacement. PMID- 15813172 TI - March madness-associated deep vein thrombosis. PMID- 15813173 TI - The unseen aesthetics of double-decker televisions: a Southernism. PMID- 15813174 TI - The patient's page. Pain control for children. PMID- 15813175 TI - The patient page. Symptom relief for menopause. PMID- 15813176 TI - Bioactive microbial metabolites. AB - The short history, specific features and future prospects of research of microbial metabolites, including antibiotics and other bioactive metabolites, are summarized. The microbial origin, diversity of producing species, functions and various bioactivities of metabolites, unique features of their chemical structures are discussed, mainly on the basis of statistical data. The possible numbers of metabolites may be discovered in the future, the problems of dereplication of newly isolated compounds as well as the new trends and prospects of the research are also discussed. PMID- 15813177 TI - YM-216391, a novel cytotoxic cyclic peptide from Streptomyces nobilis. I. fermentation, isolation and biological activities. AB - YM-216391, a novel cyclic peptide, was isolated from the cultured mycelium of Streptomyces nobilis JCM 4274. It was purified by solvent extraction, silica gel and ODS flash column chromatographies, followed by preparative HPLC. YM-216391 dose-dependently inhibited the growth of human cervical cancer HeLa S3 cells with an IC50 value of 14nM. YM-216391 also showed potent cytotoxic activity against a human cancer cell line panel. PMID- 15813178 TI - YM-216391, a novel cytotoxic cyclic peptide from Streptomyces nobilis. II. Physico-chemical properties and structure elucidation. AB - YM-216391, a novel cytotoxic cyclic peptide, has been isolated from the cultured mycelium of Streptomyces nobilis JCM 4274. The planar structure of YM-216391 was assigned on the basis of 1D and 2D NMR spectroscopic techniques. The absolute configuration of the amino acid residues in YM-216391 was determined by Marfey's analysis and chiral HPLC analysis of its acid hydrolysate. PMID- 15813179 TI - Total synthesis and biological activities of (+)-sulfamisterin (AB5366) and its analogues. AB - The first total synthesis of (+)-sulfamisterin (AB5366), a naturally occurring alpha-substituted alpha-amino acid derivative possessing a sulfonated hydroxy function, is described. Overman rearrangement of an allylic trichloroacetimidate derived from D-tartrate effectively generated the tetrasubstituted carbon containing a nitrogen substituent. Construction of the amino acid moiety and sulfonation of the hydroxy group, followed by deprotection completed the total synthesis, which fully confirmed the proposed absolute structure of the natural product. The possible stereoisomers of (+)-sulfamisterin and their desulfonated derivatives were also synthesized. Biological assessment of all synthetic compounds revealed that natural (+)-sulfamisterin and its 3-epimer as well as their desulfonated derivatives possessing 2S-configuration strongly inhibit the serine palmitoyl transferase both in vitro and in vivo, whereas compounds with 2R configuration were found to show much weaker inhibitory activity. PMID- 15813180 TI - A gamma-lactone form nafuredin, nafuredin-gamma, also inhibits helminth complex I. AB - Nafuredin, a delta-lactone antibiotic, is a fungal metabolite showing selective helminth NADH-fumarate reductase inhibition, and whose target had been revealed as complex I. We found that nafuredin is easily converted to nafuredin-gamma by weak alkaline treatment. The structure of nafuredin-gamma was elucidated as a gamma-lactone form of nafuredin with keto-enol tautomerism. Nafuredin-gamma shows similar complex I inhibitory activity as nafuredin, and it also possesses anthelmintic activity in vivo. PMID- 15813181 TI - Absolute configuration of kigamicins A, C and D. AB - The stereochemistry of kigamicins A (1), C (2) and D (3) were elucidated by a combination of X-ray crystallographic analysis and degradation studies. The absolute structures of kigamicins thus determined were depicted as shown in Fig. 2. PMID- 15813182 TI - Favolon B, a new triterpenoid isolated from the Chilean Mycena sp. strain 96180. AB - A new biologically active triterpenoid, favolon B (1), was isolated from fermentation broths of Mycena sp. strain 96180. Favolon B showed antifungal activities towards Botrytis cinerea, Mucor miehei, Paecilomyces variotii and Penicillium notatum. No activities were observed against bacteria and yeasts. The structure of favolon B was elucidated by spectroscopic techniques. PMID- 15813183 TI - Fungal phenalenones inhibit HIV-1 integrase. AB - A phenalenone compound, atrovenetinone methyl acetal, was isolated from a culture broth of Penicillium sp. FKI-1463 as an HIV-1 integrase inhibitor, and it showed anti-HIV activity in vitro. HIV-1 integrase inhibition and anti-HIV activity of two other natural phenalenones were also studied. Among the tested compounds, funalenone inhibited HIV-1 integrase with an IC50 value of 10 microM and showed the best selectivity (anti-HIV, IC50=1.7 microM; cytotoxicity, IC50=87 microM). PMID- 15813184 TI - The synthesis of 7-substituted-3-dinitrostyryl cephalosporins and their ability for detecting extended spectrum beta-lactamases (ESBLs). AB - We synthesized 7-substituted-3-(2,4-dinitrostyryl)cephalosporin derivatives which were Nitrocefin analogs, for detecting extended spectrum beta-lactamase (ESBL) specifically. HMRZ-86 which has carboxypropyloxyimino group on 7 aminothiazolacetamide substituent were not hydrolyzed by class A, C and D beta lactamases, but it was hydrolyzed by ESBL and metallo beta-lactamase (class B), then its color changed from yellow to red. The hydrolysis of metallo beta lactamase was inhibited by adding sodium mercapto acetic acid (SMA). Therefore HMRZ-86 is a useful chromogenic agent to detect ESBL specifically. PMID- 15813185 TI - A new antibiotic, antimycin Ag, produced by Streptomyces sp. K01-0031. AB - A new antimycin group antibiotic, antimycin A9, was isolated from a cultured broth of Streptomyces sp. K01-0031 together with antimycins A3a, A3b, A4, and A7, and flazin methyl ester. Antimycin A9 is the first antimycin having an aromatic 8 acyl residue. It showed potent nematocidal and insecticidal activities against Caenorhabditis elegans and Artemia salina, respectively. It inhibited bovine heart NADH oxidase at nanomolar level like other known antimycins. PMID- 15813186 TI - Macrolides with promotive activity of monocyte to macrophage differentiation. PMID- 15813187 TI - [The role of orlistat in the treatment of obesity]. AB - The overweight and obesity represent severe problems for the health management system of developed countries. In the evolution of obesity, beside genetic background, the environmental factors also play important roles. In the daily routine, the majority of obese patients need drug treatment, over the diet and physical activity. Among the available medicines the inhibitors of monoamine re uptake causes dry mouth, tachycardia, sleeplessness and elevated blood pressure, therefore, due to the frequently associated obesity and hypertension many physicians avoid using these compounds. The orlistat as a selective inhibitor of pancreatic and enteral lipase enzymes impedes the absorption of the highest calorie containing nutrients, the fats exerting beneficial effects in the treatment of obesity. The abdominal bloating and diarrhea as side effects of the drug may act as an advantage in many cases, since these happen especially in those cases when the patient neglects the previously suggested low fat diet and therefore the drug induced diarrhea and bloating may mean a feed-back for the patient in respect of the proper diet. Recent studies show many beneficial biochemical changes in obesity related pathological metabolic processes during the administration of orlistat. The authors, in their present work review in short the role of orlistat in the treatment of slimming cure. PMID- 15813188 TI - [Changes in the management of acute pancreatitis as related to its pathogenesis]. AB - Although acute pancreatitis runs a benign self limiting course in 80% of cases, acute necrotizing form of it still remained a severe disease associated with significant morbidity and mortality. Severity assessment thus plays an important role in identifying patients with high risk of local and/or systemic complications. Locally, development of necrosis especially if it becomes infected accounts for high mortality, but systemic inflammatory response syndrome (SIRS) and multiple organ failure (MOF) following necrosis further increases the risk of fatal outcome. Several scoring systems, contrast-enhanced CT scan can help to recognize patients requiring early intensive management. Prophylactic systemic antibiotic treatment and nasojejunal feeding improves prognosis by decreasing the gut derived infection of necrosis. CT guided fine needle aspiration sample must be cultured to detect infection. Conservative therapy should be continued while necrosis remains sterile, but surgical and/or CT guided percutaneous catheter drainage is mandatory when infected necrosis developed. Results of therapeutic influence on the proinflammatory cytokine cascade in acute pancreatitis are still controversial. Enteral feeding seems to be the only proven tool in attenuating acute phase response and improving disease severity. PMID- 15813189 TI - [Prognosis in acute digitalis poisoning]. AB - INTRODUCTION: Acute digitalis overdosage is characterized by high electric instability, its mortality may reach 10-15 percent even nowadays. PURPOSES: To detect the possible risk factors which might predict severe intoxication. METHODS: Data of 50 patients treated at authors' department with acute digoxin poisoning over the past 8 years could be retrospectively evaluated. Cases were classified according to the Poison Severity Score (PSS). The following parameters were taken into consideration: age, sex, diseases influencing the severity of intoxication, dose of the drug, heart frequency, serum potassium and digoxin levels and vomiting. For statistical analysis a Kruskal-Wallis test and a chance quotient calculation was applied. RESULTS: From 50 patients 30 were mild (PSS 1, 2), 20 were severely poisoned, which subgroup included 8 deaths (PSS 4) and 12 patients who recovered (PSS 3). Based on Kruskal-Wallis test significant differences were found in the following items: greater number of primary diseases PSS 4 vs other subgroups (p < 0.05); bradycardia PSS 4 vs PSS 2 (p < 0.05) and PSS 3 vs PSS 2 (p < 0.05); hyperkalaemia PSS 3 vs PSS 2 (p < 0.01); elevated serum digoxin level PSS 3 vs PSS 2 (p < 0.05). The risk of severe poisoning (PSS 3-4) was increased in case of hyperkalaemia, bradycardia, vomiting (p < 0.001), and if the patients' age and if the drug dose exceeded 65 years or 10 mg, respectively (p < 0.05). CONCLUSIONS: The predictive risk factors concerning severe acute digoxin poisoning are profuse vomiting, hyperkalaemia and bradycardia. The predictive risk factors of fatal outcome are age over 65 years associated with primary disease, vomiting and bradycardia. PMID- 15813190 TI - [Postural tachycardia syndrome]. AB - Erect posture could be a source of diverse chronic symptoms usually referred as orthostasis intolerance. Increasing number of patients suffering from orthostatic syndromes is recognized. Characterization and distinction of these orthostatic syndromes are based on the clinical presentation; including the typical history of orthostasis intolerance, and the unique hemodynamic responses on assuming upright posture. The diagnosis of postural tachycardia syndrome (POTS) requires orthostatic heart rate acceleration in excess of 120 beat/min, or an absolute increase of > or = 30 beat/min, in the absence of significant orthostatic hypotension. Subjects with POTS often exhibit more widespread symptoms of autonomic dysregulation. The diagnosis of inappropriate sinus tachycardia (IST) is based on the presence of resting tachycardia defined by pulse rate of > or = 100 beat/min, or a heart rate acceleration to same range in response to minimal stress, or activity. For the illustration of the difficulties in differentiation between these overlapping syndromes the authors present a case with POTS. The unique features of the presented case are the extreme orthostatic tachycardia (170/min), and the unexpected therapeutic effect of sinus node radiofrequency modification. We suggest a relationship between the peak orthostatic heart rate and the therapeutic value of sinus node modification. PMID- 15813191 TI - [Treatment of cardiomyopathies]. PMID- 15813192 TI - Observing and engaging in purposeful actions with objects influences estimates of their size. AB - A ladle was recalled as being taller by participants who observed tedious removal of sand from it with a small teaspoon than by those who observed removal with a larger spoon. A second experiment showed that the number of darts thrown in order to hit a target correlated negatively with memory estimates of the size of the target, a finding replicated in a third experiment with size estimates made while the target was visible. The first two experiments suggest that the way an object is used can influence memory of its size. The third experiment supports the hypothesis that in vivo size estimation of familiar objects may employ a mechanism that derives size from memory and that size memory can be distorted by the way an object was used. PMID- 15813193 TI - Nothing compares 2 views: change blindness can occur despite preserved access to the changed information. AB - Change blindness, the failure to detect visual changes that occur during a disruption, has increasingly been used to infer the nature of internal representations. If every change were detected, detailed representations of the world would have to be stored and accessible. However, because many changes are not detected, visual representations might not be complete, and access to them might be limited. Using change detection to infer the completeness of visual representations requires an understanding of the reasons for change blindness. This article provides empirical support for one such reason: change blindness resulting from the failure to compare retained representations of both the pre- and postchange information. Even when unaware of changes, observers still retained information about both the pre- and postchange objects on the same trial. PMID- 15813194 TI - Transfer of calibration in length perception by dynamic touch. AB - Earlier studies suggested that the calibration of actions is functionally, rather than anatomically, specific; thus, calibration of an action ought to transfer to actions that serve the same goal (Rieser, Pick, Ashmead, & Garing, 1995). In the present study, we investigated whether the calibration of perception also follows a functional organization: If one means of detecting an information variable is recalibrated, are other means of detection recalibrated as well? In two experiments, visual feedback was used to recalibrate perceived length of a rod wielded by the right hand; the recalibration was found to transfer to length perception with the left hand. This implies that calibration in perception is organized functionally rather than anatomically, and supports the general view that calibration applies to functional systems. PMID- 15813195 TI - Location cuing and response time distributions in visual attention. AB - The allocation of visual attention was investigated in two experiments. In Experiment 1 (n = 24), a peripheral cue was presented, and in Experiment 2 (n = 24), a central cue was used. In both experiments, cue validity was 90%, and the task was four-choice target identification. Response time distributions were collected for valid trials over five cue-target stimulus onset asynchronies (SOAs), and ex-Gaussian parameters were extracted. In both experiments, only the mean of the Gaussian component decreased as a function of cue-target SOA, which implied a strict time axis translation of the distributions. The results were consistent with sequential sampling models featuring a variable delay in the onset of information uptake. PMID- 15813196 TI - Task- and location-switching effects on visual attention. AB - In two experiments, we examined the effects of task and location switching on the accuracy of reporting target characters in an attentional blink (AB) paradigm. Single-character streams were presented at a rate of 100 msec per character in Experiment 1, and successive pairs of characters on either side of fixation were presented in Experiment 2. On each trial, two targets appeared that were either white letters or black digits embedded in a stream of black letter distractors, and they were separated by between zero and five items in the stream (lags 1-6). Experiment 1 showed that report of the first target was least accurate if it immediately preceded the second target and if the two targets were either both letters or both digits (task repetition cost). Report of the second target was least accurate if one or two distractors intervened between the two targets (the U-shaped AB lag effect) and if one target was a letter and the other a digit (task switch cost). Experiment 2 added location uncertainty as a factor and showed similar effects as Experiment 1, with one exception. Lag 1 sparing (the preserved accuracy in reporting the second of two targets if the second immediately follows the first) was completely eliminated when the task required attention switching across locations. Two-way additive effects were found between task switching and location switching in the AB paradigm. These results suggests separate loci for their attentional effects. It is likely that the AB deficit is due mainly to central memory limitations, whereas location-switching costs occur at early visual levels. Task-switching costs occur at an intermediate visual level, since the present task switch involved encoding differences without changes in stimulus-response mapping rules (i.e., the task was character identification for both letters and digits). PMID- 15813197 TI - Dynamical trajectories in category learning. AB - Category learning has traditionally been studied by examining how percentage correct changes with experience (i.e., in the form of learning curves). An alternative and more powerful approach is to examine dynamical learning trajectories--that is, to examine how the parameters that describe the current state of the model change with experience. We describe results from a new experimental paradigm in which empirical-learning trajectories are directly observable. In these experiments, participants learned two categories of spatial position, and they were constrained to identify and use a linear decision bound on every trial. The dependent variables of principal interest were the slope and the intercept of the bound used on each trial. Data from two experiments supported the following conclusions. (1) Gradient descent provided a poor description of the empirical trajectories. (2) The magnitude of changes in decision strategy decreased with experience at a rate that was faster than that predicted by gradient descent. (3) Learning curves suffered from substantial identifiability problems. PMID- 15813198 TI - Perception of place-of-articulation information in natural speech by monkeys versus humans. AB - Four monkeys and 6 humans representing five different native languages were compared in the ability to categorize natural CV tokens of /b/ versus /d/ produced by 4 talkers of American English (2 male, 2 female) in four vowel contexts (/i, e, a, u/). A two-choice "left/right" procedure was used in which both percentage correct and response time data were compared between species. Both measures indicated striking context effects for monkeys, in that they performed better for the back vowels /a/ and /u/ than for the front vowels /i/ and /e/. Humans showed no context effects for the percentage correct measure, but their response times showed an enhancement for the /i/ vowel, in contrast with monkeys. Results suggest that monkey perception of place of articulation is more dependent than human perception on the direction of the F2 onset transitions of syllables, since back-vowel F2s differentiate /b/ and /d/ more distinctively. Although monkeys do not provide an accurate model of the adult human in place perception, they may be able to model the preverbal human infant before it learns a more speech-specific strategy of place information extraction. PMID- 15813199 TI - Serial processing in melody identification and the organization of musical semantic memory. AB - Unlike the visual stimuli used in most object identification experiments, melodies are organized temporally rather than spatially. Therefore, they may be particularly sensitive to manipulations of the order in which information is revealed. Two experiments examined whether the initial elements of a melody are differentially important for identification. Initial exposures to impoverished versions of a melody significantly decreased subsequent identification, especially when the early exposures did not include the initial notes of the melody. Analyses of the initial notes indicated that they are differentially important for melody identification because they help the listener detect the overall structure of the melody. Confusion errors tended to be songs that either were drawn from the same genre or shared similar phrasing. These data indicate that conceptual processing influences melody identification, that phrase-level information is used to organize melodies in semantic memory, and that phrase level information is required to effectively search semantic memory. PMID- 15813200 TI - Oculomotor correlates of context-guided learning in visual search. AB - Previous studies have shown that context-facilitated visual search can occur through implicit learning. In the present study, we have explored its oculomotor correlates as a step toward unraveling the mechanisms that underlie such learning. Specifically, we examined a number of oculomotor parameters that might accompany the learning of context-guided search. The results showed that a decrease in the number of saccades occurred along with a fall in search time. Furthermore, we identified an effective search period in which each saccade monotonically brought the fixation closer to the target. Most important, the speed with which eye fixation approached the target did not change as a result of learning. We discuss the general implications of these results for visual search. PMID- 15813201 TI - Some characteristics of auditory spatial attention revealed using rhythmic masking release. AB - The tuning of auditory spatial attention with respect to interaural level and time difference cues (ILDs and ITDs) was explored using a rhythmic masking release (RMR) procedure. Listeners heard tone sequences defining one of two simple target rhythms, interleaved with arhythmic masking tones, presented over headphones. There were two conditions, which differed only in the ILD of the tones defining the target rhythm: For one condition, ILD was 0 dB and the perceived lateral position was central, and for the other, ILD was 4 dB and the perceived lateral position was to the right; target tone ITD was always zero. For the masking tones, ILD was fixed at 0 dB and ITDs were varied, giving rise to a range of lateral positions determined by ITD. The listeners' task was to attend to and identify the target rhythm. The data showed that target rhythm identification accuracy was low, indicating that maskers were effective, when target and masker shared spatial position, but not when they shared only ITD. A clear implication is that at least within the constraints of the RMR paradigm, overall spatial position, and not ITD, is the substrate for auditory spatial attention. PMID- 15813202 TI - Bimodal and trimodal multisensory enhancement: effects of stimulus onset and intensity on reaction time. AB - Manual reaction times to visual, auditory, and tactile stimuli presented simultaneously, or with a delay, were measured to test for multisensory interaction effects in a simple detection task with redundant signals. Responses to trimodal stimulus combinations were faster than those to bimodal combinations, which in turn were faster than reactions to unimodal stimuli. Response enhancement increased with decreasing auditory and tactile stimulus intensity and was a U-shaped function of stimulus onset asynchrony. Distribution inequality tests indicated that the multisensory interaction effects were larger than predicted by separate activation models, including the difference between bimodal and trimodal response facilitation. The results are discussed with respect to previous findings in a focused attention task and are compared with multisensory integration rules observed in bimodal and trimodal superior colliculus neurons in the cat and monkey. PMID- 15813203 TI - The mysteries of the diagonal: gender-related perceptual asymmetries. AB - In this article, we report a perceptual asymmetry for the two diagonals that is related to gender in that females prefer the diagonal spanning from top right to bottom left (/) whereas males prefer the opposite (?). This relationship is observed in a variety of tasks, including aesthetic judgment of paintings, spotting differences between two paintings, and visual search for a tilted line among similarly tilted distractors. This article does not provide an explanation of the relationship between this asymmetry and gender but rules out several potential mediating factors, such as eye dominance, head tilt, handedness, and hemispheric differences. At the same time, the scope of the phenomenon is outlined: The asymmetry is found for both meaningful and meaningless stimuli and in both brief and extended presentations. Moreover, the asymmetry is found to be related to the tilt of the visual elements that require processing, not to their location in the visual field. PMID- 15813204 TI - Rapid serial visual distraction: task-irrelevant items can produce an attentional blink. AB - When two sequential targets (T1 and T2) are presented within about 600 msec, perception of the second target is impaired. This attentional blink (AB) has been studied by means of two paradigms: rapid serial visual presentation (RSVP), in which targets are embedded in a stream of central distractors, and the two-target paradigm, in which targets are presented eccentrically without distractors. We examined the role of distractors in the AB, using a modified two-target paradigm with a central stream of task-irrelevant distractors. In six experiments, the RSVP stream of distractors substantially impaired identification of both T1 and T2, but only when the distractors shared common characteristics with the targets. Without such commonalities, the distractors had no effect on performance. This points to the subjects' attentional control setting as an important factor in the AB deficit and suggests a conceptual link between the AB and a form of nonspatial contingent capture attributable to distractor processing. PMID- 15813205 TI - Cardiologist recruitment becomes strategic, Part I: Feathering the nest. PMID- 15813206 TI - The reason for the cardiovascular test. AB - When diagnosis codes are reported and sequenced on an insurance claim, official guidelines are available to provide instructions for the correct assignment of ICD-9-CM codes. These instructions are intended to ensure that the correct reason for the diagnostic service is reported and that procedures are reimbursed when they are medically necessary. PMID- 15813207 TI - Carotid artery stenting: anatomy of a Medicare coverage decision. AB - The Centers for Medicare and Medicaid Services has made an important reimbursement decision regarding carotid artery stenting for patients at high risk for carotid endarterectomy surgery. This article will identify the carotid coverage questions and process as a case study of the issues that the Centers and stakeholders can face regarding complex new technologies, as well as the questions for carotid artery stenting and other new technologies that the process raises for your hospital. PMID- 15813208 TI - Positioning your catheterization laboratory to survive the push for noninvasive diagnostics. AB - The use of noninvasive imaging modalities such as computerized tomography and magnetic resonance for coronary disease diagnosis will dramatically decrease the volume of diagnostic patients in the cardiac catheterization laboratory. Consideration must be given to what can be done to keep the cardiac catheterization laboratory a productive and cost-effective department once diagnostics are removed from the catheterization laboratory setting. PMID- 15813209 TI - Anti-fertility effects of embelin in female Sprague-Dawley rats may be due to suppression of ovarian function. AB - Effects of embelin on oestrous cycle, plasma levels of progesterone and oestradiol, and in vitro production of oestradiol and progesterone by mixed ovarian cells was studied. Forty adult (4 months old) regularly cycling female Sprague-Dawley rats were divided into four groups of 10 rats each. Groups I and II (controls) were given 1 ml/kg body weight of physiological saline or corn oil (vehicle). Groups III and IV received 10 mg/kg and 20 mg/kg body weight embelin in corn oil, respectively. Emberlin disrupted the oestrous cycles in Groups III and IV animals, and there was a significant depression in plasma oestradiol (p <0.05) and progesterone (p <0.02) at both 10 and 20 mg/kg body weights, respectively. Isolated mixed ovarian cells from embelin treated rats produced significantly less progesterone and estradiol than controls in vitro. It is concluded that embelin probably interferes with reproductive functions in female rats by suppressing ovarian production of sex steroid hormones. PMID- 15813210 TI - Seasonal changes in plasma dehydro-epiandrosterone (DHEA) levels of domestic geese. AB - Changes in plasma DHEA, testosterone (T) and 17-B-oestradiol (E2) levels were examined in domestic geese of both sexes in the fall and winter. The levels of steroid hormones were determined in blood plasma by means of radio-immunoassay (RIA). A so-called second (autumn) cycle was induced in geese via a dark-room preparation and natural keeping conditions. The plasma levels of DHEA showed a minor peak at onset of the autumn breeding and a major one prior to the more intense spring reproduction in both sexes. The seasonal curves of plasma DHEA appeared fairly similar in ganders and layers and without considerable differences between the absolute values. In ganders, plasma DHEA peaks preceded the elevations in T levels in the fall and spring alike. With layers, in turn, the autumn and spring peaks of plasma DHEA appeared after the peaks in E2 levels. With ganders, the concentration of plasma T seemed to predominate between the two androgens throughout the experimental period. With layers, in turn, the concentration of DHEA surpassed the level of plasma E2 at the time of the peak periods and other times during the study, as well. In domestic geese, DHEA is probably involved in the autumn physiological processes and the induction of reproduction during fall and early spring periods, alike. PMID- 15813211 TI - Antinociceptive action of aqueous extract of the leaves of Ixora coccinea. AB - The aim of this study was to examine the antinociceptive potential of leaves of Ixora coccinea (family: Rubeaceae). One of four doses (500, 750, 1000 or 1500 mg/kg, n = 8/dose) of aqueous leaf extract (ALE) or 1 ml of distilled water was orally administered to male rats and antinociceptive activity was ascertained using three models ofnociception (tail flick, hot plate and formalin tests). The results showed that ALE possesses considerable antinociceptive activity (when evaluated in hot plate and formalin test but not in tail flick test). The antinociceptive activity of the ALE had a rapid onset (within 1h) and a fairly long duration of action (up to 5 h) with a peak effect at 3 h. Further, the antinociceptive activity was dose-dependent and was not associated with harmful side-effects or toxicity even following subchronic administration. The antinociceptive action was mediated centrally at the supraspinal level mainly via dopaminergic mechanism. In addition, it is likely that antioxidant activity of the ALE could have played an auxiliary role in inducing antinociception. Dopaminergic and antioxidative activities of ALE could arise, respectively, from its quaternary base alkaloid and flavonoid constituents. PMID- 15813212 TI - Autonomic nerves terminating on microvessels in the pineal organs of various submammalian vertebrates. AB - In earlier works we have found that in the mammalian pineal organ, a part of autonomic nerves--generally thought to mediate light information from the retina- form vasomotor endings on smooth muscle cells of vessels. We supposed that they serve the vascular support for circadian and circannual periodic changes in the metabolic activity of the pineal tissue. In the present work, we investigated whether peripheral nerves present in the photoreceptive pineal organs of submammalians form similar terminals on microvessels. In the cyclostome, fish, amphibian, reptile and bird species investigated, autonomic nerves accompany vessels entering the arachnoidal capsule and interfollicular meningeal septa of the pineal organ. The autonomic nerves do not enter the pineal tissue proper but remain in the perivasal meningeal septa isolated by basal lamina. They are composed of unmyelinated and myelinated fibers and form terminals around arterioles, veins and capillaries. The terminals contain synaptic and granular vesicles. Comparing various vertebrates, more perivasal terminals were found in reptiles and birds than in the cyclostome, fish and amphibian pineal organs. Earlier, autonomic nerves of the pineal organs were predominantly investigated in connection with the innervation of pineal tissue. The perivasal terminals found in various submammalians show that a part of the pineal autonomic fibers are vasomotoric in nature, but the vasosensor function of some fibers cannot be excluded. We suppose that the vasomotor regulation of the pineal microvessels in the photosensory submamalian pineal--like in mammals--may serve the vascular support for circadian and circannual periodic changes in the metabolic activity of the pineal tissue. The higher number of perivasal terminals in reptiles and birds may correspond to the higher metabolic activity of the tissues in more differentiated species. PMID- 15813213 TI - Stereological analysis of thyroid mast cells in rats after exposure to extremely low frequency electromagnetic field and the following "off" field period. AB - Influence of extremely low frequency electromagnetic field (ELF-EMF) on thyroid gland mast cells was investigated on male Mill Hill rats. Animals were exposed to EMF (50 Hz, 50 microT to 500 microT, 10 V/m) from 24 hours after birth, 7 hours/day, 5 days/week for three months when a part of animals (group I) was sacrificed, while the rest of them were subjected to recovery evaluation and sacrificed after one (group II), two (group II) and three (group IV) weeks following the exposure. Stereological analysis on toluidine blue-stained paraffin sections showed increased volume density of degranulated mast cells in all groups and, except in group III, and numerical density as well, implicating the sensitivity of thyroidal mast cells to power frequency EMFs. Since in our previous investigations, morphofunctional alterations of thyroid gland in rats exposed to ELF-EMF were found the contribution of released mast cell mediators to these changes could be presumed. PMID- 15813214 TI - Investigation of chromosome abnormalities and early embryonic mortality in goose lines. AB - Early embryonic mortality and chromosome abnormalities were studied in three goose lines: Grey Landes (line 7), White Polish (line 4) and their synthetic line (line 9). Eggs laid at the beginning, in the middle and at the end of the laying season were set. At candling at 5th day after egg set, all eggs (2847) were examined and those showing no normal embryonic development were opened 2847. Dead embryos were classified phenotypically and karyotyped. The mean ratio of embryonic mortality (EM) among fertile eggs was 9.4%, 5.2%, 7.3% in the lines 4, 7 and 9, respectively. The mean ratio of embryos with chromosomal abnormalities (CA) among the dead embryos was 8.0%, 14.8% and 13.1% in the lines 4, 7 and 9, respectively. Gander effect and layer within gander effect on embryo mortality were significant, indicating genetic factors. Father and mother of the layer effects were also significant, showing family effects. Animals producing dead embryos and embryos with chromosome abnormalities in high proportion were selected. In the selected groups the mean EM was 17.7-22.9%, and the mean CA was 11.7-34.7% among the three lines. The repetition of CA was not observed in the reproductive season of following year, while animals repeated the high EM (repeatability coefficient of 0.54). This shows that some part of EM may be resulted from other genetic factors. Ganders and layers progeny of these selected animals showed also high EM. It was concluded that culling pairs giving high EM value in their embryos could increase the average level of embryo viability and that the study of genetic determinism of that trait should be continued in geese. PMID- 15813215 TI - Transgenic mice, carrying an expressed anti-HIV ribozyme in their genome, show no sign of phenotypic alterations. AB - Transgenic mice are suitable model animals for testing the in vivo functionality of custom-tailored ribozymes. Transgenic experiments can demonstrate whether a ribozyme is able to cleave any RNA transcript of the host animal or not. Most probably, this kind of cleavage activity gives rise to phenotypic alterations in mice. In the present paper we demonstrate that an anti-HIV ribozyme does not cause any detectable phenotypic effect in mice carrying and expressing it. Our transgenic mice developed well and were indistinguishable from their wild type counterparts. PMID- 15813216 TI - Comparison on DNA patterns of different ecotypes of European corn borer (Ostrinia nubilalis Hubner) in Hungary. AB - For a molecular genetic study on Hungarian populations of European corn borer L5 stage larvae were collected from 14 places of three different regions of the country (uni- and bivoltine ecotypes). Additionally, the study included larvae from Egypt, too (multivoltine ecotype). Molecular examinations of European corn borer larvae using the study of mitochondrial cytochrome b (cyt b) revealed that by single strand conformation polimorphism (SSCP) the populations found in Hungary represented the same haplotype. Even the Egyptian sample showed no genetic divergence. Some minor deviatons were found in the case of a sample from Szekkutas, but that did not prove the genetic divergence of the bivoltine ecotype either, since the other samples of South-East Hungary did not display this kind of genetic variation. On the basis of our investigations it can be said that the univoltine and bivoltine generations, have uniform genetic complements. PMID- 15813217 TI - Cold-induced accumulation of protein in the leaves of spring and winter barley cultivars. AB - Electrophoretic pattern and quantitative changes in soluble proteins were determined in the leaves of spring and winter cultivars of barley (Hordeum vulgare L., cv. Makouei and cv. Reyhan, respectively) exposed to 4 degrees C for 14 d. Seedlings were grown in a controlled growth chamber for 2 weeks at a constant air temperature of 20 degrees C and then transferred to constant 4 degrees C for 14 d followed by returning to 20 degrees C (cold treatment), or they were maintained throughout at 20 degrees C during the experimental period of 40 d (control treatment). Plants were sampled every 48 h for leaf fresh weight measurements. Total leaf soluble proteins were extracted and their concentration was either determined by a colorimetric method, or size-fractionated on SDS-PAGE. Low temperature-induced increases in protein amount occurred over the second week of exposure to cold treatment irrespective of cultivar: the winter cultivar was 2 d prior in this response. The protein patterns and their density showed differences between-cultivars and between-temperature treatments. A new cold induced polypeptide was recognized in the leaves of winter barley cultivar on day 22 (8 d at 4 degrees C) compared to the control. This polypeptide was produced earlier over the first 48 h of low temperature in the winter cultivar compared with the spring one, recognizing in the leaves of cold-treated seedling until day 26. This more rapid response to a low temperature by the winter barley cultivar indicates a more sensitive response compared with the spring barley, probably cold-shock protein is a component of this cold-induced response. PMID- 15813218 TI - Reduction of paraquat toxicity in maize leaves by benzyladenine. AB - The protective effect of a cytokinin benzyladenine (BA), against toxicity of paraquat (PQ), a widely used herbicide and a well-known oxidative stress inducer, was investigated in the leaves of maize. Maize leaves have been pretreated with BA at concentrations of 1, 10 and 100 microM and afterwards treated with PQ. At all concentrations tested, BA retarded PQ-induced decreases in chlorophyll, carotenoid and ascorbic acid contents. Pretreatment with 10 and 100 microM of BA significantly increased superoxide dismutase (SOD) activity after 8 h of PQ treatment but there was no significant change in SOD activity in the leaves pretreated with BA at 12 and 24 h. However, peroxidase activity significantly increased in 100 microM of BA pretreated leaves. Results indicate that pretreatment with BA reduce PQ toxicity and BA-treated plants might become more tolerant against oxidative stress. PMID- 15813219 TI - A preliminary study on ultra high frequency electromagnetic fields effect on black locust chlorophylls. AB - Chlorophylls were quantitatively studied in the leaves of black locust (Robinia pseudoacacia L.) seedlings exposed to electromagnetic fields of high frequency. Exposure system was designed and built up to make possible simultaneous exposure of seedling lots (3 months old) to low power density electromagnetic fields corresponding to a frequency of 400 MHz. After three weeks of daily exposures (1, 2, 3 and 8 hours), chlorophyll levels were measured using adequate spectral device. Statistical analysis of experimental results was performed by means of t test to identify significant modifications induced by electromagnetic treatment in exposed samples in comparison to the control. Chlorophyll-a as well as chlorophyll-b level was found to decrease except the exposure time of two hours, where a considerable enhancement was noticed. It was revealed that the ratio of the two main types of chlorophyll was decreasing logarithmically to the increase of daily exposure time. PMID- 15813220 TI - The effects of fungicide benomyl (benlate) on growth and mitosis in onion (Allium cepa L.) root apical meristem. AB - In this study, the effects of benomyl, a systemic fungicide were investigated in the mitotic cell division in onion (Allium cepa) root tip cells during germination. For this aim, different concentrations (1, 2, 5, 10 and 20 mM) of benomyl solutions were used. All the concentrations used caused several abnormalities in mitotic cell divisions and the mitotic frequency in the onion root tip cells decreased as the concentration of benomyl solution increased. Based on our findings, it is reported that benomyl has some negative effects on mitotic divisions in onion root tip cells. PMID- 15813221 TI - Determination of ploidy levels in Ipheion uniflorum (R. C. Graham) Rafin (Liliaceae). AB - In this study, chromosome number and ploidy levels of Ipheion uniflorum cv. "Wisley Blue" (spring starflower) were determined. In meristematic root tip cells, chromosome number was found as 2n = 12 and 4n = 24. The ratios of diploid and tetraploid cells were found as 80.74% and 19.26%, respectively. In differentiated root tissues and mature leaf tissues ploidy levels were analysed by flow cytometry and polysomaty were found in both organs. In differentiated root tissues, ploidy levels were found as 2C, 4C, 8C and 16C DNA. In root tissues percentages of 2C, 4C, 8C and 16C nuclear DNA content were observed as 57.2%, 33.1%, 2.47% and 7.23%, respectively. In mature leaf tissues, ploidy levels were determined 2C, 4C, 8C and 16C DNA. In this tissue the frequency of 4C DNA was found very higher (74.3%) and 2C DNA content was determined as 19.2%. In mature leaf tissue, 8C and 16C nuclear DNA contents were observed as 2.72% and 3.78%, respectively. When nuclear DNA contents in leaves and roots were compared, an apparent difference in 2C and 4C DNA contents was found. PMID- 15813222 TI - Optimization of conditions for the efficient production of mutan in streptococcal cultures and post-culture liquids. AB - The strain Streptococcus sobrinus CCUG 21020 was found to produce water-insoluble and adhesive mutan. The factors influencing both stages of the mutan production, i.e. streptococcal cultures and glucan synthesis in post-culture supernatants were standardized. The application of optimized process parameters for mutan production on a larger scale made it possible to obtain approximately 2.2 g of water-insoluble glucan per 11 of culture supernate--this productivity was higher than the best reported in the literature. It was shown that some of the tested beet sugars might be successfully utilized as substitutes for pure sucrose in the process of mutan synthesis. Nuclear magnetic resonance analyses confirmed that the insoluble biopolymer synthesized by a mixture of crude glucosyltransferases was a mixed-linkage (1-->3), (1-->6)-alpha-D-glucan (the so-called mutan) with a greater proportion of 1,3 to 1,6 linkages. PMID- 15813223 TI - Flocculation of Bacillus thuringiensis var. israelensis suspension and its efficacy against mosquito larvae. AB - Bacillus thuringiensis ssp. israelensis (Bti) is increasingly used as an ecologically friendly anti-mosquito agent. The bacterium cells undergo fermentation in dilute suspensions; before practical use, therefore it is necessary to concentrate the suspensions. Aggregation by polymers is a powerful tool with which to regulate the stability of suspensions. Typically, polymers at low concentrations destabilize and at high concentrations stabilize colloidal systems. Bti suspensions can be flocculated efficiently by either cationic or anionic polyelectrolytes. Cationic polyelectolytes were found to be the most efficient flocculants for bacterial suspensions. It was shown that the degree of toxicity of the flocculated Bti suspensions for biting mosquito larvae was in the same range than in non-flocculated suspension. PMID- 15813225 TI - The testis histology of artificially maturated European eel (Anguilla anguilla L.) at the end of sexual maturation, and spermatozoa ultrastructure in freshwater rearing. Short communication. AB - The artificial induction of sexual maturation of European eel males was carried out by using weekly hCG administrations. Histological pictures showed that the testis tissues developed and regressed naturally and no pathological changes took place under the conditions of artificial rearing in freshwater. According to light and electron microscopic investigations the morphology and motility of the spermatozoa of males kept in freshwater proved to be similar to those in seawater. The authors suppose that freshwater rearing of males is not a barrier factor in the artificial propagation of European eels. PMID- 15813224 TI - Effect of different dietary fat sources on production traits, lipid peroxide status and on the glutathione redox system in African catfish [Clarias gariepinus (Burchell)] fingerlings. Short communication. AB - Lipids are used to provide the energy to cover the metabolic needs and to provide essential fatty acids, which are important for membrane function [12]. Fats may contain high level of long chain polyunsaturated fatty acids, which are prone to peroxidation [8] and will interact with the antioxidant defense system [1]. There is contradiction in the literature about whether the intake of fish oil enhance [7] or deplete [4] tissue antioxidant defenses and the glutathione redox system in different organisms. The aim of the present study was to examine the effects of different dietary oils on parameters of the lipid peroxide state and the glutathione redox system in C. gariepinus fingerlings. PMID- 15813226 TI - Cryopreservation of European eel (Anguilla anguilla) sperm using different extenders and cryoprotectants. Short communication. AB - Experiments were carried out on the sperm cryopreservation of artificially induced eels. The effects of several extenders and two cryoprotectants on the motility of spermatozoa were investigated. The highest post-thaw motility was observed with the combination of Tanaka's extender and DMSO as cryoprotectant. Further dilution after thawing resulted in complete loss of motility in samples frozen in presence of DMSO while sperm frozen with methanol as cryoprotectant retained its motility after further dilution. PMID- 15813227 TI - Making technology work for you. PMID- 15813228 TI - Ethics, rights, and responsibilities (RI). PMID- 15813229 TI - Health care alternatives. New concepts focus on prevention to decrease the cost of health services. PMID- 15813230 TI - The American College Health Association National College Health Assessment (ACHA NCHA), Spring 2003 Reference Group report. AB - Assessing and understanding the health needs and capacities of college students is paramount to creating healthy campus communities. The American College Health Association-National College Health Assessment (ACHA-NCHA) is a survey instrument developed by the American College Health Association (ACHA) in 1998 to assist institutions of higher education in achieving this goal. The ACHA-NCHA contains approximately 300 questions assessing student health status and health problems, risk and protective behaviors, access to health information, impediments to academic performance, and perceived norms across a variety of content areas, including injury prevention; personal safety and violence; alcohol, tobacco, and other drug use; sexual health; weight, nutrition, and exercise; and mental health. Twice a year, ACHA compiles aggregate data from institutions using the ACHA-NCHA to provide a reference group for data comparison. A portion of the data from the Spring 2003 Reference Group is provided in this article for use by professionals, researchers, institutions, departments, and organizations invested in advancing the health of college students. PMID- 15813231 TI - Sexually transmitted disease services at US colleges and universities. AB - The authors' objectives in this study were to describe the proportion of schools providing and the percentage of students with access to HIV and sexually transmitted disease (STD) education, treatment, and prevention services at 2-year and 4-year US colleges and universities. The authors mailed self-administered questionnaires to a stratified random sample (n = 910) of the 2,755 US schools with an enrollment of more than 500 students; 736 (81%) returned the survey. Four hundred seventy-four schools (60%) had a health center, representing 73% of students. Schools with a health center or housing for students were more likely to provide STD education; 52% of the schools made condoms available to students. Sixty percent of schools with health centers could test for both Chlamydia trachomatis and Neisseria gonorrhoeae, but only 67% of these schools screened women for these infections. Although most schools provided some prevention education, access to prevention, testing, and education should be increased at schools where these services are possible but not available. PMID- 15813232 TI - Factors associated with undertreatment of medical student depression. AB - The authors measured factors associated with under-treatment of medical students' depression. They administered a cross-sectional Beck Depression Inventory and sociodemographic questionnaire to students at 1 medical school, defining their outcome measure as the use of counseling services or antidepressant medication. Of an estimated 450 available student participants in the study, 322 (71.6%) completed the questionnaire. Forty-nine students (15.2%) were classified as depressed and 10 (20.4%) reported experiencing suicidal ideation during medical school, but only 13 (26.5%) of the depressed students reported treatment. The researchers observed no difference in treatment by year in school, completion of psychiatric requirement, race, or depression severity. Treatment for depression was significantly associated with older age and personal and family histories of depression. Despite the availability of effective medications and confidential mental health services, medical students with depression are undertreated. The authors' findings support the need for targeted messages to help medical students recognize their depression and refer themselves for appropriate treatment. PMID- 15813233 TI - Men's perceptions and knowledge of human papillomavirus (HPV) infection and cervical cancer. AB - The authors assessed young men's knowledge and perceptions of genital human papillomavirus (HPV) infection to identify factors that predict intention to make positive behavioral changes. Male university students aged 18 to 25 years completed a self-report instrument to assess knowledge and perceptions of genital HPV infection. If diagnosed with HPV, most men (95%) reported that they would use condoms with new partners. The intention to reduce number of sex partners was associated with an understanding that HPV may have severe consequences for women, whereas intention to encourage female sex partners to undergo Pap smear screening was associated with increased general knowledge of HPV infection. The authors concluded that it is important to include men in HPV education and prevention efforts, especially within the context that HPV may lead to cervical cancer in female partners. PMID- 15813234 TI - Development and validation of the College Tobacco Survey. AB - The authors report on the development and assessment of an instrument to measure baseline campus cigarette use and outcomes from prevention programs, including those using a social norms approach combined with environmental policy change. They administered the 37-item College Tobacco Survey (CTS) to a convenience sample of 1,279 college students in freshmen-level classes at a large urban university. Factor analysis of 15 belief items revealed 3 factors: Peer Environment, Personal Effects, and Campus Policy Endorsement. The findings support the survey's reliability and validity. The authors discuss potential uses of the survey in terms of social norms and environmental prevention programs. PMID- 15813235 TI - The impact of the erosion of retiree health benefits on workers and retirees. PMID- 15813236 TI - Rate hikes trending down, but 2005 increases still in double digits. PMID- 15813237 TI - Pay-for-performance study offers benchmarks on feedback, payment trends. PMID- 15813238 TI - Profits roll for fully capitated IPA, but red flags loom. PMID- 15813239 TI - Medicaid program saves big with prior authorization for proton pump inhibitors. PMID- 15813240 TI - Obesity redux: beware health care costs of obese members aging in place. PMID- 15813241 TI - The renoprotective potential of pentoxifylline in chronic kidney disease. AB - Current interventions with proven efficacy, such as glycemic and blood pressure control, dietary protein restriction, and angiotensin II blockade, slow the progression of chronic kidney disease (CKD); however, whether long-term cessation of CKD progression is possible remains unclear. Because of the pathogenetic complexity of this condition, multidrug interventions with the least adverse effects should be investigated as the next step in attempts to stop CKD progression. Pentoxifylline, a non-selective phosphodiesterase inhibitor with indiscernible toxicity, exerts potent inhibitory effects against cell proliferation, inflammation, and extracellular matrix accumulation, all of which play important roles in CKD progression. Pentoxifylline monotherapy markedly reduces proteinuria in patients with membranous nephropathy. Moreover, limited human studies have proven pentoxifylline efficacy in reducing proteinuria in patients with diabetes receiving angiotensin-converting enzyme inhibitors, and in patients with nephrotic syndrome secondary to lupus nephritis despite immunosuppressive therapy. Further clinical trials are necessary to examine whether pentoxifylline can improve renal outcomes in patients receiving interventions of proven efficacy. PMID- 15813242 TI - Clinical findings, treatment and prognosis in patients with severe acute respiratory syndrome (SARS). PMID- 15813243 TI - Prevalence and correlates of depressive symptoms in older immigrants (from Taiwan and mainland China) in Canada. PMID- 15813244 TI - Clinical characteristics, management and prognostic factors in patients with probable severe acute respiratory syndrome (SARS) in a SARS center in Taiwan. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is an emerging viral infectious disease. We report our experience in treating SARS patients. METHODS: From April 27 to May 24, 2003, a total of 36 patients with probable SARS were admitted and treated in a hospital rearranged as a special center for the management of SARS patients. Medical records for the patients were retrospectively reviewed. Univariate and multivariate analyses were performed to determine factors associated with respiratory failure and intubation. RESULTS: Of the 36 patients with probable SARS (median age, 37 years; range, 22-66 years), 9 were male and 27 were female. Thirty-two patients (88.9%) were infected in the hospital setting. All patients presented with fever, and 33 eventually developed lymphopenia during hospitalization. Chest radiography showed no unique pattern, but pleural effusion was not seen. All patients initially received empiric antibacterial therapy against common causative pathogens of atypical pneumonia. Ribavirin was given to all except 1 patient. Twenty-two patients received immunoglobulin therapy, and 32 were given corticosteroids. A total of 20 patients (55.6%) required supplemental oxygen, and 8 (22.2%) were intubated with mechanical ventilatory support. Two of these patients died. A higher body temperature at presentation (median 39.5 vs 38.6 degrees C), and higher peak values of lactate dehydrogenase (410 vs 282 U/L) and C-reactive protein (10.2 vs 2.5 mg/dL), were associated with subsequent respiratory failure. Multivariate analysis showed that peak level of C-reactive protein was the only independent predictor of respiratory failure and intubation (odds ratio for every increment of 1 mg/dL = 1.45; 95% confidence interval = 1.003, 2.097; p = 0.048). CONCLUSION: All patients with probable SARS who were admitted to hospital presented with fever and lymphopenia. While the efficacy of different treatments could not be evaluated from this retrospective study, a higher value of C reactive protein was associated with the development of respiratory failure and subsequent intubation. PMID- 15813245 TI - Prevalence and correlates of depressive symptoms in older Taiwanese immigrants in Canada. AB - BACKGROUND: There is a lack of research regarding depression in older Taiwanese immigrants in North American countries. This study in Canada therefore examined the prevalence of depressive symptoms among older immigrants from Taiwan, and psychosocial factors as predictors of depressive symptoms reported by older Taiwanese immigrants. METHODS: Ninety-eight migrants (aged > or = 55 years) from Taiwan to Canada, who were part of a multi-site study of health and well-being in a total of 2,272 older ethnic Chinese individuals in community dwellings, completed a face-to-face interview and answered questions in an orally administrated questionnaire. Depressive symptoms were measured by a Chinese version of the Geriatric Depression Scale. RESULTS: Of the 98 migrants from Taiwan, 21.5% reported at least a mild level of depression. Predictive factors for depressive symptoms were a negative attitude towards aging, poor general physical health, single marital status, barriers in terms of gaining access to health care services, poor financial status, lower level of identification with Chinese health beliefs, and low income. CONCLUSION: The prevalence of depressive symptoms in older Taiwanese immigrants in Canada was higher than that reported by older adults in the general Canadian population. Thus, implications for the delivery of health care services, and possible strategies to enhance the mental well-being of older Taiwanese immigrants, are discussed. PMID- 15813246 TI - Civilian gunshot wounds to the brain. AB - BACKGROUND: Civilian gunshot wounds to the brain are relatively rare, and a much neglected subject in Taiwan. We present our experience with 16 patients who sustained gunshot wounds to the brain, and then identify factors determining the respective outcomes. METHODS: From 1988-2002, data from 16 patients with civilian gunshot wounds to the brain, who were treated at Taichung Veterans General Hospital, were retrospectively reviewed. Historical information, clinical manifestations, and imaging findings were described. Based on Glasgow Outcome Score (GOS), patients were divided into a poor-outcome group (GOS, 1-2) and satisfactory-outcome group (GOS, 3-5) for further analysis. RESULTS: The overall mortality rate was 31.3% (5 of 16 patients). Thirteen patients underwent surgery, and the surgical mortality rate was 15.4% (2 of 13 patients). Of the 7 patients with a Glasgow Coma Scale (GCS) score of more than 8, all survived with satisfactory outcomes; of the 9 patients with a GCS score of less than 8, 8 had poor outcomes (5 died, and 3 were in a persistent vegetative state); p < 0.005 between the 2 GCS groups. In addition, the rate of satisfactory outcome was significantly higher in 7 patients with limited brain injury, as determined by computed tomography (CT) scan, than in 8 patients with extensive brain injury (86% vs 25%; p < 0.05). CONCLUSION: GCS score on admission, and the extent of brain injury as visualized by CT scan, seem to be the 2 most significant predictors of outcome in cranio-cerebral gunshot wounds. Patients with a GCS score of more than 8, or brain lesions limited to a single lobe of the brain, may benefit from aggressive management. PMID- 15813247 TI - Detection of subarachnoid hemorrhage at acute and subacute/chronic stages: comparison of four magnetic resonance imaging pulse sequences and computed tomography. AB - BACKGROUND: Acute subarachnoid hemorrhage (SAH) has traditionally been diagnosed by computed tomography (CT); however, fluid-attenuated inversion recovery (FLAIR) is a magnetic resonance imaging (MRI) modality currently used to detect acute SAH. CT is insensitive in the detection of subacute or chronic SAH. The purpose of this study was to compare 4 MRI pulse sequences and CT in the detection of SAH in acute and subacute-to-chronic stages. METHODS: From 2001-2003, we collected data for 22 patients (12 men and 10 women, aged 35-80 years) with SAH due to ruptured aneurysm (n = 11), trauma (3), or unknown origin (8). All patients underwent MRI and CT examination, with an interval of less than 12 hours between the 2 procedures. We divided patients into 2 groups according to the time from symptom onset to MRI evaluation: patients with MRI performed < or = 5 days post ictus had acute-stage illness, whereas patients with MRI performed from day 6-30 post-ictus had a subacute-to-chronic condition. MRI (1.5-T) pulse sequences comprised spin-echo T1-weighted, fast spin-echo T2-weighted, FLAIR, and gradient echo (GE) T2*-weighted images. RESULTS: In the acute-stage group, SAH was seen as an area of high signal intensity compared with surrounding cerebrospinal fluid in 36.4% of cases on T1-weighted images, and in 100% on FLAIR images; low signal intensities were seen in 18.2% of cases on T2-weighted images, and in 90.9% on GE T2*-weighted images. High-attenuated SAH was seen on CT in 90.9% of cases. FLAIR (p = 0.008), GE T2*-weighted images (p = 0.012) and CT images (p = 0.012) were all statistically significant indicators of acute SAH. In the subacute/chronic stage group, SAH was detected on T1-weighted images (36.4% of cases), FLAIR (33.3%), T2-weighted images (9.1%), GE T2*-weighted images (100%), and CT (45.5%). GE T2*-weighted images were significantly superior (p = 0.001) to other MRI pulse sequences and CT as indicators of subacute-to-chronic SAH. CONCLUSION: FLAIR and GE T2* MRI pulse sequences, and CT scans, are all statistically significant indicators of acute SAH. GE T2*-weighted images are statistically significant indicators of subacute-to-chronic SAH, whereas other MRI pulse sequences, and CT scans, are not. PMID- 15813248 TI - Arterial misplacement of a femoral central venous catheter complicated with acute arterial occlusion. AB - Femoral vein catheterization is often carried out during resuscitation and in critical care units. It is usually achieved via a blind, external landmark-guided technique, through manual localization of the femoral artery. However, this approach can be challenging in patients with severe shock because of absence or ambiguity of the arterial pulse. We report a case of inadvertent cannulation, with a large-bore catheter, of the right femoral artery, which was mistaken as a venous route for medication and massive transfusion. The large cannula caused direct mechanical obstruction, while intra-arterial medications induced vascular injury and vasospasm. Both factors led to thrombosis and occlusion of the right external iliac artery, thus jeopardizing the distal blood supply, and ultimately resulting in muscle necrosis of the involved limb, and the need for thrombectomy and extensive fasciotomy to salvage the extremity. This case highlights that correct placement of a central venous catheter should be clearly ascertained before the catheter is used for medical treatment, especially when catheterization is performed in shock status. PMID- 15813249 TI - Localized amyloid deposition in the nasopharynx and neck, mimicking nasopharyngeal carcinoma with neck metastasis. AB - Amyloidosis results from the deposition of amyloid proteins in organs and tissues. Clinically, it can be classified into systemic and localized forms. Here, we report a case of localized amyloidosis of the nasopharynx and neck. The initial presentation was a nasopharyngeal mass, and bilateral neck masses, mimicking nasopharyngeal carcinoma with neck metastasis. Computed tomographic scans of the neck revealed asymmetry between the bilateral nasopharyngeal walls, and multiple radio-opaque masses in both sides of the neck. A nasopharyngeal biopsy was performed and confirmed amyloid deposition. Subsequent neck-mass excision biopsies confirmed that the neck masses were also amyloid deposits. Further laboratory examinations revealed no systemic involvement. There was no disease progression after local excision. Localized amyloidosis in the head and neck is rare, but can have various manifestations that may sometimes mimic neoplasms. PMID- 15813250 TI - Intramural gallbladder hematoma mimicking gallbladder neoplasm in a 33-year-old male. AB - We report a case of intramural gallbladder hematoma mimicking gallbladder neoplasm in a 33-year-old male; bleeding was confined to the gallbladder wall without rupture into the lumen. The patient presented with sudden onset of right upper quadrant pain, with no history of abdominal trauma. The abdomen was soft, with mild tenderness over the right upper quadrant, and a positive Murphy's sign. Abdominal sonography, computed tomography, and endoscopic retrograde cholangiopancreatography all revealed the presence of a mass lesion in the gallbladder wall. Thus, a gallbladder tumor was highly suspected. Laparoscopic cholecystectomy was performed, and intramural hematoma was found intraoperatively. The cause of intramural hemorrhage was obscure. PMID- 15813251 TI - Laparoscopic assessment and treatment of non-palpable testis in an 18-year-old male. AB - Laparoscopy, both diagnostic and therapeutic, has been used in the management of pediatric non-palpable testes. We report a case of left non-palpable testis at the unusual age of 18 years. Laparoscopic exploration revealed an intra-abdominal testis lying between the internal inguinal ring and external iliac vessels. The testis was visually in good condition and, in contrast to the accepted procedure of orchiectomy performed in late adolescent cryptorchidism, we decided to preserve this intra-abdominal testis, which was located too far from the scrotum for a primary orchidopexy. The first stage of the Fowler-Stephens procedure was performed laparoscopically. The second-stage orchidopexy was performed successfully through an inguinal approach 3 months later, after sonographic ascertainment of non-decreased testicular volume. Laparoscopy allows thorough exploration for an intra-abdominal testis, with simultaneous therapeutic options. Our experience highlights the feasibility of laparoscopic assessment and treatment of cryptorchidism in adolescents and young adults. PMID- 15813252 TI - Wide-range tunable Fabry-Perot array filter for wavelength-division multiplexing applications. AB - A new design for a Fabry-Perot-based array filter for wavelength-division multiplexing (WDM) applications is proposed. The device is composed of two partially reflective mirrors; the surface of one of the mirrors is stepwise. Not only does the configuration of multiple air-gap cavities into a single unit provide wavelength tunability but the channel numbers can be expanded. A four channel multiplexer has been demonstrated. The results revealed that a compact and cost-effective WDM device should be easily attainable. PMID- 15813253 TI - White-light photorefractive phase mask. AB - A volume photorefractive phase mask has been fabricated with incoherent white light from an incandescent source in LiNbO3:Fe self-defocusing photorefractive crystal for the first time, to our knowledge. It can guide and modulate a probe white light or a laser beam and can be used to transmit an incoherent dark image as the guided modes of the waveguides induced by white-light dark spatial solitons. This also proves the existence of photorefractive nonlinearity of white light. PMID- 15813254 TI - Photolithographic fabrication of a high-precision bar-code pattern on the surface of a sphere. AB - Bar-code patterns were produced upon gold-coated glass spheres 8 mm in diameter with a novel projection photolithography exposure system that included a computer controlled rotation stage-and-shutter arrangement. The patterns extended in an equatorial band about the entire 360 degrees periphery of the sphere. To obtain uniform thickness of the photoresist layer, we dip coated the spheres and removed the excess with an absorption pad. The widths of the bars in the pattern were in the range 21.85 +/- 0.6 microm at the equator, and the relative error in the angular position of the pattern features was less than 0.025 degrees. The patterned spheres are suitable for use in optically interrogated flying spot magnetic heading sensors. PMID- 15813255 TI - Comparative analysis of techniques for measuring the modulation transfer functions of charge-coupled devices based on the generation of laser speckle. AB - Two methods for measuring the modulation transfer function (MTF) of a charge coupled device (CCD) that are based on the generation of laser speckle are analyzed and compared. The method based on a single-slit aperture is a quick method, although the measurements are limited to values of less than the Nyquist frequency of the device. The double-slit method permits the measurement of values of as much as some 1.8 times the Nyquist frequency, although it is a slower method because of the necessity to move the CCD. The difference between the MTF values obtained with the two methods is less than 0.1 in magnitude; the root-mean square error between the two curves is 0.046 (4.6%). PMID- 15813256 TI - Temperature imaging in nonpremixed flames by joint filtered Rayleigh and Raman scattering. AB - Joint fuel Raman and filtered Rayleigh-scattering (FRS) imaging is demonstrated in a laminar methane-air diffusion flame. These experiments are, to our knowledge, the first reported extension of the FRS technique to nonpremixed combustion. This joint imaging approach allows for correction of the FRS images for the large variations in Rayleigh cross section that occur in diffusion flames and for a secondary measurement of fuel mole fraction. The temperature-dependent filtered Rayleigh cross sections are computed with a six-moment kinetic model for calculation of major-species Rayleigh-Brillouin line shapes and a flamelet-based model for physically judicious estimates of gas-phase chemical composition. Shot averaged temperatures, fuel mole fractions, and fuel number densities from steady and vortex-strained diffusion flames stabilized on a Wolfhard-Parker slot burner are presented, and a detailed uncertainty analysis reveals that the FRS-measured temperatures are accurate to within +/- 4.5 to 6% of the local absolute temperature. PMID- 15813257 TI - Measurement of focal length with phase-shifting Talbot interferometry. AB - Phase-shifting interferometry with a Fourier fringe analysis technique is implemented to analyze Talbot interferometric fringes and to evaluate the focal length of a lens. A four-step algorithm is used to obtain the phase map of the lens. The slope of the phase map is related to the focal length, and it is from this relationship that we evaluate the focal length. Experimental results are presented. Our experimental study suggests that phase-shifting Talbot interferometry combined with a Fourier fringe analysis technique can be advantageously used to improve the accuracy of measurement. PMID- 15813258 TI - Two-wavelength phase-shifting interferometry with a superimposed grating displayed on an electrically addressed spatial light modulator. AB - A two-wavelength moire phase-shifting interferometer that uses a superimposed grating has been developed. The optical phase shifts for the two wavelengths are given by digital phase shifts of a superimposed grating displayed on a liquid crystal spatial light modulator. A phase shift of the moire fringe is achieved by equal phase shifts with opposite signs in the two gratings. A moire phase shifting interferometer with no moving parts and no requirement for calibration of the value of the phase shifts was obtained. Our experimental result shows measurements of the profile of a step object with a 2.65-microm synthetic wavelength. PMID- 15813259 TI - Investigation of the influence of spatial coherence of a broad-area laser diode on the interference fringe system of a Mach-Zehnder interferometer for highly spatially resolved velocity measurements. AB - Laser Doppler anemometry is a method for absolute velocity measurements that is based on a Mach-Zehnder interferometer arrangement and usually employs transverse fundamental-mode lasers. We employed inexpensive and powerful broad-area laser diodes and investigated ways in which an interference fringe system is influenced by the spatial coherence properties of a multimode beam. It was demonstrated that, owing to poor spatial coherence of the beam, interference is suppressed in the marginal regions of the intersection volume. Based on these results, a sensor for highly spatially resolved velocity measurements can be built. The inherent astigmatism of the broad-area diode is corrected by an arrangement of two crossed cylindrical lenses. An interference fringe system of length 200 microm and a relative variation in fringe-spacing of only 0.22% were demonstrated with light emitted from a broad-area laser diode with a 100 microm x 1 microm emitter size. Based on this principle a powerful, simple, and robust laser Doppler sensor has been achieved. Highly spatially resolved measurements of a boundary layer flow are presented. PMID- 15813261 TI - Nanoengineered liquid mirrors shaped by thermal fields. AB - We discuss a new type of deformable mirror made from nanoengineered reflective layers deposited onto liquids. The surfaces are shaped by heating with a laser. The response times of the deformed surfaces are slow (> 1 s). Simplicity and low cost appear to be the main advantages of thermally deformable liquid mirrors. PMID- 15813260 TI - In vivo scattering measurement of biological tissue by the use of a pyroelectric polymer transducer. AB - We propose a new method, to our knowledge, for noninvasive scattering measurements of tissues by the use of a pyroelectric polymer transducer, poly(vinylidene fluoride trifluoroethylene) film. In this method, samples are irradiated with nanosecond, low-energy light pulses delivered from an optical fiber, and the pyroelectric signal induced by the diffuse reflectance is measured with a transducer. The signal is then converted into diffuse reflectance by use of a calibration factor. The validity of this method was confirmed by our measuring the scattering coefficient of a white acrylic resin within an accuracy of +/- 15%. We attempted to apply this method to the estimation of scattering coefficients of normal and burned skins in rats in vivo. PMID- 15813262 TI - Controlled frustration of total internal reflection by electrophoresis of pigment particles. AB - A model based on geometrical optics has been developed to describe the photometric observations associated with a novel method to control the reflectance of a surface. In this new reflectance modulation approach, electrophoresis of pigment particles is used to absorb light reflected by total internal reflection (TIR). The pigment particles are sufficiently small that they substantially do not scatter light, but rather they modify the effective refractive index at the reflection interface. An incident light ray interacting with this modified effective index is attenuated in a spectrally selective manner. Although frustrated TIR has been understood and used in various applications for some time, in this case it is used to substantially modify the color of the reflected light, which to our knowledge has not been previously reported. A numerical model of the pigment particle distribution has been developed to describe the observations. PMID- 15813263 TI - Far-field intensity distribution of a beam generated by a resonator with a phase unifying mirror. AB - The far-field intensity distribution (FFID) of a beam generated by a phase unifying mirror resonator was investigated based on scalar diffraction theory. Attention was paid to the parameters, such as obscuration ratio and reflectivity of the phase-unifying mirror, that determine the FFID. All analyses were limited to the TEM00 fundamental mode. PMID- 15813264 TI - Comparison of relative signal-to-noise ratios of different classes of imaging spectrometer. AB - The continued development of new and fundamentally different classes of imaging spectrometer has increased both the scope and the complexity of comparisons of their relative signal-to-noise ratios. Although the throughput and multiplex advantages of Fourier-transform spectrometers were established in the early 1950s, the application of this terminology to imaging spectrometers is often ambiguous and has led to some confusion and debate. For comparisons of signal collection abilities to be useful to a system designer, they must be based on identical requirements and constraints. We present unambiguous definitions of terminology for application to imaging spectrometers and comparisons of signal collection abilities and signal-to-noise-ratios on a basis that is useful to a systems designer and inclusive of six fundamentally different classes (both traditional and novel) of imaging spectrometers. PMID- 15813265 TI - Computer-generated holograms with optimum bandwidths obtained with twisted nematic liquid-crystal displays. AB - We discuss a computer-generated hologram for encoding arbitrary complex modulation based on a commercial twisted-nematic liquid-crystal display. This hologram is implemented with the constrained complex modulation provided by the display in a phase-mostly configuration. The hologram structure and transmittance are determined to obtain on-axis signal reconstruction, maximum bandwidth, optimum efficiency, and high signal-to-noise ratio. We employed the proposed holographic code for the experimental synthesis of first-order Bessel beams. PMID- 15813266 TI - GaAs-based multiple-quantum-well spatial light modulators fabricated by a wafer scale process. AB - The design, fabrication, and characterization of large, two-dimensional multiple quantum-well modulator arrays are presented. Such arrays present a speed advantage compared with competing technologies such as liquid crystals and micromirrors, which are intrinsically limited to the kilohertz range. We discuss the design compromises to reach high-contrast, low-voltage swing optical structures compatible with complementary metal-oxide semiconductor-based integrated circuits and present experimental results. Contrast ratio of 5:1 (limited by the fill factor), variations in uniformity below 1nm, and frame rates in excess of 10kHz are demonstrated. Technology maturity for volume production is also discussed. PMID- 15813267 TI - Unique measurement of the parameters of a twisted-nematic liquid-crystal display. AB - The parameters of a twisted-nematic liquid-crystal display (TN-LCD) are measured with no ambiguity when we measure the intensity transmittance of a system that simply consists of a TN-LCD, two polarizers, a quarter-wave plate, and a monochromatic beam. First we show analytically that the director angle can have a 90 degree ambiguity and the twist angle of the liquid-crystal molecules a 180 degree ambiguity. Then we uniquely measure the parameters by fitting the theoretical predictions to the intensity transmittance measured with and without the quarter-wave plate and by using the quasi local-adiabatic condition. The birefringence of the TN-LCD is measured next as a function of the applied voltage. We design a phase spatial light modulator by using the measured parameters and measure the phase delay of the output beam to excellent agreement with the theoretical prediction. PMID- 15813268 TI - Application of the symplectic finite-difference time-domain method to light scattering by small particles. AB - A three-dimensional fourth-order finite-difference time-domain (FDTD) program with a symplectic integrator scheme has been developed to solve the problem of light scattering by small particles. The symplectic scheme is nondissipative and requires no more storage than the conventional second-order FDTD scheme. The total-field and scattered-field technique is generalized to provide the incident wave source conditions in the symplectic FDTD (SFDTD) scheme. The perfectly matched layer absorbing boundary condition is employed to truncate the computational domain. Numerical examples demonstrate that the fourth-order SFDTD scheme substantially improves the precision of the near-field calculation. The major shortcoming of the fourth-order SFDTD scheme is that it requires more computer CPU time than a conventional second-order FDTD scheme if the same grid size is used. Thus, to make the SFDTD method efficient for practical applications, one needs to parallelize the corresponding computational code. PMID- 15813269 TI - Particle extinction measured at ambient conditions with differential optical absorption spectroscopy. 1. system setup and characterization. AB - We describe an instrument for measuring the particle extinction coefficient at ambient conditions in the spectral range from 270 to 1000 nm. It is based on a differential optical absorption spectroscopy (DOAS) system, which was originally used for measuring trace-gas concentrations of atmospheric absorbers in the ultraviolet-visible wavelength range. One obtains the particle extinction spectrum by measuring the total atmospheric extinction and subtracting trace-gas absorption and Rayleigh scattering. The instrument consists of two nested Newton type telescopes, which are simultaneously used for emitting and detecting light, and two arrays of retroreflectors at the ends of the two light paths. The design of this new instrument solves crucial problems usually encountered in the design of such instruments. The telescope is actively repositioned during the measurement cycle. Particle extinction is simultaneously measured at several wavelengths by the use of two grating spectrometers. Optical turbulence causes lateral movement of the spot of light in the receiver telescope. Monitoring of the return signals with a diode permits correction for this effect. Phase sensitive detection efficiently suppresses background signals from the atmosphere as well as from the instrument itself. The performance of the instrument was tested during a measurement period of 3 months from January to March 2000. The instrument ran without significant interruption during that period. A mean accuracy of 0.032 km(-1) was found for the extinction coefficient for an 11-day period in March. PMID- 15813270 TI - Use of optical scattering to discriminate particle types in coastal waters. AB - The particulate scattering characteristics of coastal waters were examined at nine locations around the United States, including near-shore sites in the Gulf of Mexico and the Atlantic and Pacific oceans. The scattering measurements were used in conjunction with inversion models to estimate particle size distributions and bulk refractive indices of the suspended particles. The relationships between various scattering properties and chlorophyll concentration were also investigated and compared with previous relationships described for case I waters. Although the general patterns of scattering and particle characteristics in coastal waters were fairly consistent, fine-scale variability within the water column was substantial. Combining optical measurements with inversion techniques provided a more informative view of the environment and a better understanding of the nature of particle populations in the coastal ocean. PMID- 15813271 TI - Direct spectral measurements with a Brewer spectroradiometer: absolute calibration and aerosol optical depth retrieval. AB - We present three different methods for the absolute calibration of direct spectral irradiances measured with a Brewer spectroradiometer, which are shown to agree to within +/- 2%. Direct irradiance spectra derived by Brewer and Bentham spectroradiometers agree to within 4 +/- 3%. Good agreement was also found by a comparison of the aerosol optical depth and Angstrom exponent retrieved by the two instruments and a multifilter rotational shadowband radiometer. The spectral aerosol optical depth (300-365 nm) derived from six years of direct irradiance measurements at Thessaloniki shows a distinct seasonal variation, averaging to approximately 0.3 at 340 nm in winter and approximately 0.7 in summer. PMID- 15813272 TI - Tunable waveguides via photo-oxidation of plasma-polymerized organosilicon films. AB - Plasma-enhanced chemical vapor deposition (PECVD) of dimethylsilane and hexamethyldisilane produced thin films with a refractive index of 1.56 +/- 0.01 at 633 nm. A decrease in the refractive index of approximately 3% was observed after irradiation with UV light from an ArF laser operating at 193 nm. Lower intensity UV light from a Hg arc lamp induced a slower and controllable decrease in the refractive index. Top-side prism coupling showed the as-deposited organosilicon films to be multimode at 633 nm and single mode at 1550 nm. A model predicted that 30 s of UV irradiation with the Hg arc lamp would decrease the refractive index of the light-guiding film by approximately 0.01, converting the waveguide into single-mode operation across the spectrum of essential wavelengths for microphotonics. Irradiation followed by further coupling experiments confirmed this tunability. Trimming the refractive index of patternable organosilicon polymeric films presents a method of optimizing the coupling performance of PECVD microphotonic interconnect layers postdeposition. PMID- 15813273 TI - Bend loss effects in diffused, buried waveguides. AB - Bend loss effects can be a significant concern in the design and performance of diffused, buried waveguide devices. Since diffused, buried waveguides typically do not have analytical mode solutions, the bend mode must be expressed as an expansion of straight waveguide modes. For the case of buried ion-exchanged waveguides, the bend loss is affected by bend radius, the duration of the ion exchange and burial processes, as well as the size of the mask opening used to create the waveguides and applied field during burial. The bend loss effects for each of these variables are explored under typical fabrication conditions. PMID- 15813274 TI - Diode-pumped Yb,Er:glass laser passively Q switched with a V3+:YAG crystal. AB - Saturable-absorber Q switching of a diode-pumped erbium-doped glass laser at 1.54 microm with a V:YAG crystal has been demonstrated. An average output power of 30 mW with Q-switching efficiency of 37% and pulse duration of 350 ns have been obtained. Results of the numerical simulation of the laser have been presented. PMID- 15813275 TI - Two-channel self-mixing laser Doppler measurement with carrier-frequency-division multiplexing. AB - We demonstrate real-time two-channel self-mixing laser-Doppler measurement with extreme optical sensitivity using a laser-diode-pumped thin-slice LiNdP4O12 laser. Successful carrier-frequency-division-multiplexed two-channel operations are realized by using one laser, two sets of optical frequency shifters, and a two-channel frequency-modulated-wave demodulation circuit. Simultaneous independent measurements of vibrations of speakers and averaged motions of small Brownian particles in different scattering cells are demonstrated. Self-mixing photon correlation spectroscopy of particle size distributions is also discussed. PMID- 15813276 TI - Compact, passively Q-switched Nd:YAG laser for the MESSENGER mission to Mercury. AB - A compact, passively Q-switched Nd:YAG laser has been developed for the Mercury Laser Altimeter, an instrument on the Mercury Surface, Space Environment, Geochemistry, and Ranging mission to the planet Mercury. The laser achieves 5.4% efficiency with a near-diffraction-limited beam. It passed all space-flight environmental tests at subsystem, instrument, and satellite integration testing and successfully completes a postlaunch aliveness check en route to Mercury. The laser design draws on a heritage of previous laser altimetry missions, specifically the Ice Cloud and Elevation Satellite and the Mars Global Surveyor, but incorporates thermal management features unique to the requirements of an orbit of the planet Mercury. PMID- 15813277 TI - Thermal dephasing in second-harmonic generation of an amplified copper-vapor laser beam in beta barium borate. AB - The conversion efficiency in second-harmonic generation of an amplified beam in a master-oscillator power amplifier copper-vapor laser (CVL) is lower than that of the oscillator beam alone. This lower efficiency is often vaguely attributed to wave-front degradation in the amplifier. We investigate the role of wave-front degradation and thermal dephasing in the second-harmonic generation of a CVL from a beta-barium borate crystal. Choosing two beams with constant intrapulse divergence, one from a generalized diffraction filtered resonator master oscillator alone and other obtained by amplifying oscillator by use of a power amplifier, we show that at low flux levels the decrease in efficiency is due to wave-front degradation. At a fundamental power above the critical power for thermal dephasing, the decrease is due to increased UV absorption and consequent thermal dephasing. Thermal dephasing is higher for the beam with the lower coherence width. PMID- 15813278 TI - Effects of a nonuniform vertical profile of chlorophyll concentration on remote sensing reflectance of the ocean. AB - Numerical simulations of radiative transfer were used to examine the effects of a nonuniform vertical profile of the inherent optical properties of the water column associated with the vertical profile of chlorophyll concentration, Chl(z), on the spectral remote-sensing reflectance, Rrs(gamma), of the ocean. Using the Gaussian function that describes the Chl(z) profile, we simulated a relatively broad range of open-ocean conditions characterized by the presence of a subsurface Chl maximum at depths greater than or equal to 20 m. The simulations for a vertically nonuniform Chl(z) were compared with reference simulations for a homogeneous ocean whose Chl was identical to the surface Chl of inhomogeneous cases. The range of values for the Gaussian parameters that produce significant differences in Rrs(gamma) (> 5%) was determined. For some vertical structures of Chl(z) considered, the magnitude of Rrs(gamma) and the blue-to-green band ratios of Rrs(gamma) differ significantly from the reference values of homogeneous ocean (> 70% in extreme cases of low surface chlorophyll of 0.02 mg m(-3) and shallow pigment maximum at 20 m). The differences are small or negligible when the nonuniform profiles are characterized by a surface Chl greater than 0.4 mg m(-3) or a depth of Chl maximum greater than 45 m (65 m in extremely clear waters with a surface Chl of 0.02 mg m(-3) or less). The comparison of modeling results with the current algorithm for retrieving the global distribution of chlorophyll from satellite imagery of ocean color suggests that strong effects of the subsurface chlorophyll maximum on reflectance at low surface chlorophyll concentrations can lead to a severalfold overestimation in the algorithm-derived surface chlorophyll. Examples of field data from the Sea of Japan and the north polar Atlantic Ocean are used to illustrate various nonuniform pigment profiles and their effect on the blue-to-green ratio of Rrs(gamma). PMID- 15813279 TI - Optical system design and integration of the mercury laser altimeter. AB - The Mercury Laser Altimeter (MLA), developed for the 2004 MESSENGER mission to Mercury, is designed to measure the planet's topography by laser ranging. A description of the MLA optical system and its measured optical performance during instrument-level and spacecraft-level integration and testing are presented. PMID- 15813280 TI - Simple algorithm to determine the near-edge smoke boundaries with scanning lidar. AB - We propose a modified algorithm for the gradient method to determine the near edge smoke plume boundaries using backscatter signals of a scanning lidar. The running derivative of the ratio of the signal standard deviation (STD) to the accumulated sum of the STD is calculated, and the location of the global maximum of this function is found. No empirical criteria are required to determine smoke boundaries; thus the algorithm can be used without a priori selection of threshold values. The modified gradient method is not sensitive to the signal random noise at the far end of the lidar measurement range. Experimental data obtained with the Fire Sciences Laboratory lidar during routine prescribed fires in Montana were used to test the algorithm. Analysis results are presented that demonstrate the robustness of this algorithm. PMID- 15813281 TI - When your patients are itching to see you: not all hives are urticaria. AB - When patients present with itching and the perception that they have hives, what other processes can mimic urticaria? With the exception of urticarial vasculitis, urticaria typically lasts less than 24 to 36 hours at one site. A rash that persists longer should raise the suspicion of another inflammatory process. When the hive-like rash does not respond to antihistamines, a biopsy may reveal an alternative diagnosis. All biopsies should also be submitted for immunofluorescence to exclude atypical presentations of inflammatory bullous disease presenting with urticaria. However, even biopsies can be subject to misinterpretation and if the clinical picture does not support the biopsy, an alternative consultation with a dermatopathologist may be required. The extent of the laboratory and radiologic evaluation should be dictated by the clinician's suspicion of alternative causes for the hive(s) because rarely malignancies may present with urticaria. Common things are indeed common with urticaria and the more urticaria does not appear to be typical, the more often the clinician should consider alternative diagnoses. PMID- 15813282 TI - Approach to therapy in chronic urticaria: when benadryl is not enough. AB - Chronic urticaria is a relatively common disorder in which its management often presents a difficult clinical challenge. Histamine antagonists are the mainstays of therapy, and the second-generation antihistamines offer an attractive combination of efficacy, convenience, and lack of side effects. For patients whose symptoms are not controlled by once daily second-generation antihistamines, adjunct therapy with first generation H1-antihistamines, H2-antihistamines, doxepin, or leukotriene modifiers may be effective. Short-term systemic corticosteroids are useful for acute symptom flares but should not be used chronically. For chronically unremitting disease, immunomodulatory agents such as low-dose oral cyclosporine often are useful. PMID- 15813283 TI - Chronic urticaria in children. AB - Chronic urticaria (CU) in children often is caused by physical triggers. It also can be associated with autoimmune disease, especially thyroid autoimmunity. Functional autoantibodies against FcepsilonRIalpha have been reported in children with CU but have not been studied in a systematic fashion. The autologous serum skin test has been proposed as a surrogate test to define the presence of these autoantibodies, although it identifies the presence of histamine-releasing factor, not necessarily antibody. Second-generation antihistamines usually are preferred for chronic therapy. Leukotriene receptor antagonists may be of benefit in a subset of children with CU. PMID- 15813284 TI - Are attention deficit hyperactivity disorder and chronic fatigue syndrome allergy related? what is fibromyalgia? AB - Despite the progress made in the field of allergy-immunology in recent years, there are a group of diseases that the allergist-immunologist may be called on to manage in which their precise etiologies have not been identified but that appear to be initiated or exacerbated by allergic mechanisms. Attention deficit hyperactivity disorder (ADHD), chronic fatigue syndrome (CFS), and fibromyalgia (FM) fall into this category of disorders. Although the precise etiology of ADHD still remains unknown, the most prevalent theory is that it represents a neurobiologically based developmental disability leading to inadequate production of the neurotransmitter dopamine. In patients with CFS, there appears to be a fundamental dysfunction of the neuroendocrine-immunological system with deficiencies of immunological and neurological function, which, together with chronic viral infection, may lead to a sequence of events responsible for the symptoms of this disorder. FM appears to be a variant of CFS with a predominance of hypothalamic pituitary axis dysfunction. The disorder is characterized by chronic widespread pain and the finding of 11/18 tender points on examination. Now, there is emerging evidence to suggest that adverse reactions to foods or food components also may be associated with behavioral disturbances that may play a role in each of these disorders. An understanding of the interactive responses involved in the neuroendocrine-immunological network is essential for a comprehension of the pathophysiology of ADHD, CFS, and FM and the role of allergies appears to be an important triggering event in each of the disorders. PMID- 15813285 TI - Complying with federal guidelines: Medicare Prescription Drug, Improvement, and Modernization Act of 2003: amendments to Social Security Act. AB - The Medicare Prescription Drug, Improvement, and Modernization Act of 2003 is the most comprehensive health legislation since the initiation of Medicare. It has many other ramifications in addition to establishing a prescription drug payment. This review outlines the prescription drug benefit and attempts to extract the topics important to practicing physicians from the other eleven titles. We will be seeing the effects of this act for years to come. PMID- 15813286 TI - Rhinitis-asthma connection: epidemiologic and pathophysiologic basis. AB - Over the last several years, important research has demonstrated the link between the upper airways disease, rhinitis, and lower airways disease, asthma. In fact, it appears that asthma and rhinitis represent components of a single inflammatory airways disease. With the use of epidemiologic data and pathophysiologic studies, the connection between these inflammatory conditions becomes clear. Numerous population studies show the high rate of asthma in patients with allergic rhinitis compared to the general population. Several surveys suggest the rate of rhinitis in asthma patients to be as high as 100%. Recent studies in humans demonstrate that local allergen exposure in the nose of patients with allergic rhinitis can quickly lead to significant allergic inflammation in the lungs, even without a previous history of asthma or airways bronchial hyperreactivity. It is important for the clinician to understand the connection between these disorders so that all patients with rhinitis are evaluated for lower airways disease, and all patients with asthma be screened for upper airways conditions. PMID- 15813287 TI - Onset of efficacy of montelukast in seasonal allergic rhinitis. AB - Montelukast has shown efficacy for seasonal allergic rhinitis (SAR); however, onset of action for SAR has not been presented. The aim of this study was to determine the onset of action of montelukast, 10 mg, measured in days after starting once-daily therapy, in spring and fall studies of SAR. Data were analyzed from four 2-week, double-blind, placebo-controlled phase III clinical trials. End points included the daytime nasal symptoms score, the nighttime symptoms score, the composite symptoms score (mean of the scores for daytime nasal symptoms and nighttime symptoms), and the daytime eye symptoms score. Using a 0-3 scale (no symptoms to severe symptoms), daytime symptoms were rated by patients once daily each evening; nighttime symptoms were rated once daily each morning. Analyses of data pooled across the four evening dosing studies showed that montelukast, compared with placebo, produced significant improvement (p < or = 0.001) from baseline by day 2 of treatment (after 2 doses) in the daytime nasal symptoms score, nighttime symptoms score, and composite symptoms score. Differences between montelukast and placebo for these end points in mean change from baseline at day 2 were -0.08 (95% CI, -0.12, -0.03), -0.08 (95% CI, -0.13, 0.04), and -0.08 (95% CI, -0.12, -0.04), respectively. These data represented a mean reduction for montelukast of 11-13% in symptom scores from baseline for each end point at day 2. When compared with the full 2-week response, these observed differences for each end point at day 2 (after 2 doses) represented a substantial proportion (over 70%) of the overall treatment benefits seen. Significant improvement (p < or = 0.001) in the daytime eye symptoms score was seen by day 1 (-0.08 [-0.12, -0.03]), after the first dose. In patients treated for SAR, montelukast has a beneficial effect on daytime and nighttime symptoms by the 2nd day of daily therapy. PMID- 15813288 TI - Theophylline inhibits integrin-dependent eosinophil superoxide production. AB - Theophylline has been proposed as a drug that is able to reduce eosinophil activation in asthma. We tested the hypothesis that it can interfere with the integrin-mediated stimulation of eosinophil function. Eosinophils from healthy donors were triggered by monoclonal antibodies to beta1- and beta2-integrins in the presence of different concentrations of theophylline: 4.3 microg/mL (2.4 X 10(-5) M) 13 microg/mL (7.2 X 10(-5) M) 26 microg/mL (1.4 X 10(-4) M), and 43 microg/mL (2.4 X 10(-4) M), respectively. The level of activation was evaluated by assaying O2- generation. A statistically significant inhibition (p < 0.05) of O2- generation was observed with the different concentrations of theophylline when eosinophils were triggered via very late antigen 4 (VLA-4), lymphocyte function antigen 1 and the common beta2-chain. No effect of theophylline on O2- generation was observed in phorbol-myristate-acetate-stimulated eosinophils. These results suggest that theophylline can interfere with the eosinophil activation triggered by ligation of beta1- and beta2-integrins. This effect of theophylline possibly may play a relevant role in the inhibition of eosinophil infiltration and activation at the sites of allergic reactions. PMID- 15813289 TI - Grape anaphylaxis: a study of 11 adult onset cases. AB - Reports of immunoglobulin E (IgE)-mediated allergic reactions to grapes and wine are limited in the literature. Nevertheless, grapes are widely grown and consumed in Mediterranean countries. The object of this prospective study was to present clinical features, in vivo and in vitro allergy testing, and human leukocyte antigen (HLA) serotyping in patients with recurring reactions to grapes and grape products. Eleven unrelated Greek patients, six men and five women (aged 16-44 years; mean, 26.9 years) were enrolled based on a documented history of IgE mediated reactions to grapes, wine, or other grape products. Their evaluation included full history, reaction severity, clinical examination, skin-prick tests with food allergens and molds, serum IgE, specific IgEs to the same allergen battery, and HLA typing. Patients reported 35 grape-induced anaphylaxis episodes ranging from moderate (more than one system involved but not prominent respiratory or cardiovascular symptoms; 45.5%) to severe (serious respiratory obstruction and/or hypotension and loss of consciousness; 54.5%). A causative agent was identified: wine, 10/35 (28.6%); red grapes, 9/35 (25.7%); stuffed vine leaves, 8/35 (22.9%); raisins, 3/35 (8.6%); white grapes, 2/35 (5.7%); wine vinegar, 2/35 (5. 7%); and grape juice, 1/35 (2.9%). Other foods that induced anaphylaxis were apples (54.5%), cherries (18.6%), peaches (18.6%), and bananas (9.3%). Specific IgE values were in accordance with skin-prick tests reactivity. Concerning HLA typing, 9/11 possessed HLA-DR11(5) and -DQ7(3) and the remaining two possessed HLA-DR17(3) and -DQ2 antigens. Grapes, wine and other grape products might cause serious allergic reactions in sensitized individuals. The cosensitization and reaction incidence to other fruit allergens could be a basis for further investigation of panallergens of fruits. HLA class II antigens may contribute in genetic predisposition to these allergic reactions. PMID- 15813292 TI - Highlighting 2004 award-winning initiatives. AB - This issue takes a closer look at how five award-winning healthcare organizations are finding--and continually refining--innovative ways to provide high-quality healthcare. One of those organizations is Robert Wood Johnson University Hospital Hamilton, which recently was named the fourth healthcare winner of the annual Malcolm Baldrige National Quality Award. The Joint Commission on Accreditation of Healthcare Organizations recently selected two facilities in the hospital category--Stamford Hospital and Staten Island University Hospital--as recipients of the eighth annual Codman Award for their work in using outcomes measurement to promote quality care. The Reading Hospital and Medical Center received a Cheers Award from the Institute for Safe Medication Practices for its toolkit promoting patient safety. Sentara Healthcare System, top winner of the American Hospital Association's Quest for Quality Award, has been cited for its efforts to align its quality and safety goals with its organizational goals. PMID- 15813290 TI - Prevalence of skin test reactivity in patients with convincing, vague, and unacceptable histories of penicillin allergy. AB - Penicillin (PCN) allergy has been vastly overdiagnosed, and too many people are incorrectly labeled as allergic to PCN, which affects their health by preventing the use of beta-lactam antibiotics. This investigation explores whether taking a careful history can eliminate the need for some to carry the PCN allergy label. A retrospective study of a focused history and PCN skin testing was done in a consecutive sample of a suburban allergy population of 319 patients who had a positive history of PCN allergy. The patients were divided into three groups based on PCN history: convincing, vague, and unacceptable. The convincing group had patients with impressive histories of PCN allergy likely to be immunoglobulin E-mediated. The vague group had unimpressive but plausible histories of PCN allergy. The unacceptable group were those patients with the PCN-allergic label who were either never exposed to PCN or the PCN reaction was too far-fetched to be believable. Out of 319 patients with a positive PCN allergy history, 135 (42.3%) patients were classified as convincing, 150 (47%) patients were classified as vague, and 34 (10.7%) patients were classified as unacceptable. Positive PCN skin tests were found in 19 of 135 (14.1%) patients in the convincing group, in 10 of 150 (6.7%) patients in the vague group, and 0 of 34 (0%) patients in the unacceptable group. The finding that 6.7% of patients with a vague PCN allergy history had positive skin tests suggests that skin testing is necessary in this group. The PCN-allergic label in the unacceptable group had been unchallenged by 33 primary care physicians and 9 allergists. This study suggests that physician acceptance of unwarranted PCN-allergic labels is not uncommon, and that such labels can be removed without skin testing. PMID- 15813294 TI - Establishing rules of the road for pharmaceutical representatives. PMID- 15813293 TI - Open access: nothing this good can be easy. PMID- 15813295 TI - MOC triggers resignation. PMID- 15813296 TI - MOC implementation unfair. PMID- 15813297 TI - Grace period could make MOC less time-demanding. PMID- 15813298 TI - Days too short for MOC. PMID- 15813299 TI - Enough is enough. PMID- 15813300 TI - Asthma days worth further chronicling. PMID- 15813301 TI - Conflict of interest. PMID- 15813302 TI - EHR vendor rating tool. PMID- 15813303 TI - The value of a coding education. PMID- 15813304 TI - Should you treat yourself, family or friends? PMID- 15813305 TI - What you need to know about the Medicare Prescription Drug Act. PMID- 15813306 TI - Answers to your questions about same-day scheduling. PMID- 15813307 TI - A tool for evaluating patients with knee injury. PMID- 15813308 TI - How to cure a bad case of fax-o-rrhea. PMID- 15813309 TI - How to become a defensive filer. PMID- 15813310 TI - Illegal immigrants with ESRD: The impact on U.S. and Canadian health care facilities. PMID- 15813311 TI - DaVita sees improved quality of life when sending patients home. PMID- 15813312 TI - Linking CKD and heart disease. PMID- 15813314 TI - Heart health and kidney disease. PMID- 15813315 TI - Self control leads to a healthy life on dialysis. PMID- 15813316 TI - Q & A: ask the experts. An interview with Joseph Eustace. PMID- 15813317 TI - Tax saving approaches for your medical practice. PMID- 15813318 TI - Discussing end-of-life care or what do we know? PMID- 15813319 TI - Physician workforce: coming up short. PMID- 15813320 TI - Can physician assistants fill the gaps in the nephrology workforce? PMID- 15813321 TI - For the love of butterflies: an interview with Luz Manso, the 2004 iKidney.com essay winner. PMID- 15813322 TI - Flip a coin. PMID- 15813323 TI - Living organ donations: ethical concerns. PMID- 15813324 TI - Assessment Management of trigeminal nerve injuries in dentistry. PMID- 15813325 TI - Oral pathology quiz # 45. Case number 1. Malignant salivary gland tumor. PMID- 15813326 TI - Be aware of immigration issues when hiring. PMID- 15813327 TI - Negotiation. PMID- 15813328 TI - Mandatory reporting of child abuse and liability for inadequate staffing. PMID- 15813330 TI - Patient's perception of nurse staffing. PMID- 15813331 TI - PHCC alumnus delivers graduation address. PMID- 15813332 TI - Information technology: supporting the practice of nurses. PMID- 15813333 TI - Pioneering the environmental health frontier. PMID- 15813334 TI - [Secondary cardiovascular prevention]. PMID- 15813335 TI - [Cardiac pacing in 2005]. AB - Cardiac pacing is still evolving in 2005. "Classical" indications for cardiac pacing (sinus node dysfunction, atrio-ventricular bloc) represent the majority of indications for pacemaker implantation but technologic advances offer now new hopes for pacing in patients with advanced heart failure, vasovagal syncope or paroxysal AFib. PMID- 15813336 TI - [Resistant hypertension]. AB - Hypertension is a cardiovascular risk factor which needs a good evaluation before treatment. When this latter is decided, the target is to normalize high blood pressure. This requires a complete information of the patient; the latter will also receive individualized non pharmacological advices and also possibly different antihypertensive drugs. When blood pressure does not normalize, one must check the pressure measurement technique, the compliance to treatment and potential pharmacological interferences. Secondary hypertension is only considered if resistance to therapy cannot be found. It should be remembered that obesity and sleep apnea disorders are responsible of many instances of resistant hypertension. PMID- 15813337 TI - [Silent myocardial ischaemia: a deafening silence]. AB - Angina pectoris, whether stable or unstable, is not the most common ischaemic manifestation of coronary heart disease. Silent or asymptomatic ischaemia is the most frequently encountered ischaemic event in coronary heart disease. nevertheless, its prevalence is very variable, depending on the studied population (2-4% in asymptomatic patients, 20-30% after an myocardial infarction, 40-60% in stable angina and 60-80% in unstable angina). In this context, silent ischaemia raises many questions with regards to the rational use of diagnostic tools (stress test, holter electrocardiographic recording and stress imaging tests), their prognostic interpretation and its management. PMID- 15813338 TI - [Anti-platelet treatment in cardiology]. AB - Antiplatelets are today one of the pivotal treatment in cardiovascular disease. Aspirin, used for more than 100 years, continue to be the most used drug in the treatment of athe rosclerosis. However, new antiplatelet molecules such as clopidogrel and antibodies against glycoprotein IIb-IIIa are now part of the daily treatment for the general as well for the interventional cardiologist. This paper is a review of these different compounds in the light of the huge data available in the literature. PMID- 15813339 TI - [Innovations in the treatment of aneurysms of the ascending aorta]. AB - Numerous innovations conceming the difficult problems of ascending aortic surgery appear regularly in the literature. Two of these have been selected for this review because of the impact they have had on the treatment of aneurysms of the ascending aorta. These are the aortic valve sparing operations for root replacement and canulation of the right subclavian artery. the first is a modification of the Bentall-DeBono operation, avoiding the replacement of structurally normal valve leaflets and thus avoiding long term anticoagulation treatment. The safety of the procedure and the long term results are excellent. Subclavian canulation allows better cerebral protection when circulatory arrest is required and has simplified considerably surgery for both aortic dissections, ascending aorta and arch aneurysms. PMID- 15813340 TI - [What do we actually know about out-of-hospital cardiac arrest?]. AB - Each year at least 300,000 people in the United States and 8000 to 10,000 people in Switzerland suffer from out-of-hospital cardiac arrest, mostly due to ventricular fibrillation. Early defibrillation provides definitive treatment for most of cardiac arrest victims. Semi-automatic external defibrillators are easy to handle devices allowing to deliver an early electric shock and can be successfully used by lay people following minimal training. Newer strategies of defibrillation designed to respond faster to out-of-hospital cardiac arrest, including public access defibrillation, as well as improvement of each link of the chain of survival appears as the best strategy for the management of out-of hospital cardiac arrest. PMID- 15813341 TI - [Detection and management of early Pseudomonas aeruginosa infection in patients with cystic fibrosis]. AB - Initial Pseudomonas aeruginosa infection in patients with cystic fibrosis is a key step in pulmonary evolution of the disease. This first infection is most of the time early in life and gives some practical problems to the clinicians. It is important to look for colonisation with Pseudomonas aeruginosa as often as possible, at least every 3 month, by sputum culture when the child is able to or with induced sputum by hypertonic saline nebulisation. Treatment with inhalation of an antibiotic, preferably tobramycin for a 28 days course. Efficacy of the treatment as to be assessed by new culture. PMID- 15813342 TI - [An impossible co-existence]. PMID- 15813343 TI - [The sixth-year optional medical school curriculum]. PMID- 15813344 TI - [Protective effects of pravastatin against atherosclerosis in children with familial hypercholesterolemia]. PMID- 15813345 TI - [Looking back on Francois Mitterrand, the lies and the medical secret (I)]. PMID- 15813346 TI - [New techniques in bone and joint imaging (2)]. PMID- 15813347 TI - [Avian influenza: a risk to humans]. PMID- 15813348 TI - [The Federation Mediation Helvetica: we must not weaken now]. PMID- 15813349 TI - [Dilemmas of contemporary epilepsy pharmacotherapy]. AB - The last decade was characterized by an enormous progress in all fields of epilepsy. The availability of a relatively great number of drugs that can be categorized into standard and new antiepileptic drugs (AEDs) has made individualization of therapy possible, but has challenged the clinician to answer many questions in everyday clinical routine that are not even theoretically satisfactorily solved. Among other questions, it refers to the choice of the most appropriate AED for specific therapeutic situations. Certainly, clinical trials that are harmonized with the principles of evidence-based medicine (EBM) constitute the basis for the evaluation of AEDs. In the forming of value judgments concerning new AEDs, the most influential were clinical trials with these drugs tested as add-on therapy in pharmacoresistant epilepsy. However, many relevant clinical questions have remained unanswered by these studies, primarily because the design of these trials was more appropriate for the licensing of new drugs than for clinical practice. As far as such a complex clinical condition as epilepsy is concerned, it is difficult to define the design for clinical drug trials that might be considered generally acceptable. Nevertheless, randomized controlled studies in newly diagnosed epilepsy, where the main outcome variable is the seizure absence, constitute the main methodological standard. The disproportion between the enormous quantity of easily accessible information about epilepsy therapy and the individual capability to meaningfully process that information is certainly one of the main challenges faced by a clinician. One of the solutions offered lately by some respectable professional organizations might be the implementation of treatment guidelines made on the basis of available studies that satisfy the standards of EBM. It can be assumed that the existing guidelines, and the guidelines that are yet expected, will be very influential. However, there is a prevailing consensus that therapy should be definitely individualized. Based on the contemporary level of knowledge, it means that it should be the result of dynamic interaction between the seizure type and epileptic syndrome, specific features of AEDs, and individual characteristics of the patient. PMID- 15813350 TI - [The effect of antiepileptic drug therapy on cognitive evoked potentials and reaction time]. AB - It is well known that antiepileptic drugs have side effects on cognitive and behavioral functioning of the treated patients. AIM: Our goal was to investigate their effect on cognitive evoked potential peak latencies and patient reaction times to target stimuli for two different, acoustic and visual stimulating paradigms. Therefore, a control group of healthy subjects and three groups of patients treated with antiepileptic drugs (carbamazepine, phenobarbital) were compared. PATIENTS AND METHODS: Group 1 of 19 epileptic patients had undertherapeutic and group 2 of 16 patients had a therapeutic concentration of these drugs. Group 3 of 18 patients were treated with carbamazepine in therapeutic dose for nonepileptic reasons. The N1, P2, N2, P3a and P3b peak latencies as well as reaction time to the same target stimuli were analyzed. RESULTS AND DISCUSSION: Obtained for acoustic and visual stimulating paradigm in the control group showed no significant differences results suggesting it to be sufficient to use simple acoustic paradigm for screening. The hypothesis that antiepileptic drugs influence the parameters of cognitive evoked potentials when used as polytherapy was confirmed. The reaction time measured in the same groups showed statistically significant differences between healthy subjects and drug treated patients. The effect of antiepileptic drugs on peak latencies of cognitive evoked potentials was statistically significant, and an even greater effect was recorded on the reaction time of treated patients. CONCLUSION: The results obtained in the study were in accordance with our hypothesis that this neurophysiological method could be successfully used as a screening method to check the cognitive and motor state of antiepileptic medication. PMID- 15813351 TI - [Epileptic patient compliance with prescribed medical treatment]. AB - Medically treated patients' non-compliance with the prescribed scheme is an exceptionally important issue, faced relatively often in medical treatment of epilepsy. OBJECTIVE, PATIENTS AND METHODS: Different demographic and clinical data, collected upon questionnaire completion from 146 patients suffering from epilepsy, were sorted out and integrated in the present research analyses with the objective to determine the possible correlation of collected data and compliance level recorded in the observed patients. RESULTS AND DISCUSSION: Generally, the subjects took pretty the prescribed therapy regularly: in 62% of subjects the compliance level was recorded as good, in 23% as satisfactory, and in the remaining 15% as unsatisfactory. The following factors were identified as possible predictors of the less precise adherence to the prescribed therapeutic regimen (p<0.01): regular consumption of alcohol, medical treatment of 6 or more years, and three or more daily drug doses. The correlation between the greater number of prescribed anti-epileptic medicines and lower compliance level (p=0.05) was also assessed. The study revealed a tendency towards lower compliance level in patients aged below 30. Surprisingly, the same tendency was observed in the subjects with more thoroughly controlled epileptic condition. In spite of different experiments aimed to improve the regularity in taking the prescribed therapy, the issue of non-compliance still poses a considerable obstacle to the more successful treatment of epileptic patients. PMID- 15813352 TI - [Treatment of West syndrome]. AB - PURPOSE: West syndrome (WS) is one of the catastrophic epileptic syndromes in infancy characterized by a triad of infantile spasms, psychomotor deterioration and hypsarrhythmic EEG pattern. WS is commonly associated with poor long-term outcome, especially in symptomatic cases, with development of other seizure types, impaired cognitive and psychosocial functioning. The aim of our study was to evaluate the efficacy of the control of infantile spasms using synthetic ACTH or vigabatrin in newly diagnosed cases and to correlate it with the underlyning causes, outcome and adverse effects. PATIENTS AND METHODS: The database of children with WS seen at the Neuropediatric Unit and followed at outpatient clinics from January 1, 1994 until December 31, 2003 were reviewed. The diagnosis of WS following the criteria of ILAE was made in 32 patients. RESULTS: Data were collected for 32 children (9 girls and 23 boys). According to the etiology, 5 (15.6%) were cryptogenic, and 1 (3.1%) was idiopathic. In 26 (81.2%) symptomatic cases, hypoxic-ischemic encephalopathy (69.2%) was the most common etiologic factor, followed by central nervous system anomaly including malformation of cortical development (11.5%), and Sturge Weber syndrome (3.8%), and chromosomal translocation with Down syndrome (11.5%). In 65.1% of symptomatic cases birth occurred prematurely. The mean age at spasm onset was 5.8 months, and mean age at diagnosis and treatment 7.2 months. Between 1994 and 1996 synthetic ACTH was used for treatment of WS in 7 patients (1 cryptogenic and 6 symptomatic), spasm control was achieved in 6, hypsarrhythmia disappeared in 5, and vigabatrin was added after synthetic ACTH in 3 patients. In one child synthetic ACTH was stopped because of arterial hypertension. All children had Cushing syndrome. After 1996, vigabatrin was administrated to 5 children with cryptogenic and 20 children with symptomatic WS. In 22/32 spasm control was achieved within 15 days. Synthetic ACTH was added in 3 children with spasms and hypsarrhythmia disappeared in 1 child. There was no recurrence of WS. The mean follow-up in 27 children was 4.6 (0.5 to 9.9 years) whereas 5 were lost from follow-up. Of 6/27 children with cryptogenic WS, 1 had idiopathic WS, 3 had normal psychomotor development and 2 had psychomotor retardation, without epileptic fits and still receiving AED. Of 21/27 children with symptomatic WS 76.2% had severe psychomotor retardation, 42.8% had epilepsy, 23.8% had intractable epileptic fits, and 2 children with Down syndrome were without epilepsy and without AED. Lennox-Gastaut syndrome developed in 14.2% (3/21 children); 1 of them died at the age of 3.5 years from acute gastric bleeding during the administration of synthetic ACTH, and an other child died at the age of 5.5 years from infection and respiratory insufficiency. The mortality rate was 7.4% (2/27 children). DISCUSSION AND CONCLUSION: The cryptogenic etiology is associated with a very low risk of poor outcome in WS. In children with normal development and regular school performance an idiopathic etiology can be presumed. The children with Down syndrome had a relatively benign outcome with regard to seizure control compared with symptomatic infantile spasms in the general population. In symptomatic WS caused by hypoxic-ischemic encephalopathy the outcome was linked with coexistence of other forms of epilepsy and neurologic deficit. The poor prognosis concerning intractable nature of the seizures and serious neurologic deficit is recorded in children with malformation of cortical development and Sturge Weber syndrome. The outcome of these children is determined by the brain damage other than by epilepsy itself. Regarding the treatment with synthetic ACTH or vigabatrin, the control of WS was the same for cryptogenic and symptomatic forms, one drug may be effective if the other drug fails. Synthetic ACTH can have many side effects, even death. The visual field defect is associated with vigabatrin, but can be avoided with careful funduscopic follow-up. Vigabatrin can be suggested as the first drug for WS; if spasms persist after 15 days with a dose of 150 mg/kg, synthetic ACTH should be considered. PMID- 15813353 TI - [Epilepsy--an ongoing challenge]. AB - Epilepsy is a sequel of depolarizing neuronal event with recurrent seizures. Neuronal membrane disorder is the main cause of epileptogenesis. The epileptic process is initiated and progressing due to the membrane inability to maintain balanced changing of the electrochemical gradient, which is physiologically necessary for intracerebral signal transfer. Membrane ion channels are deranged, which may be congenital or acquired. To date, only a few specific genetically caused ion channel aberrations in some less common epileptic syndromes have been identified, however, the main pathophysiologic sequence of events at the cellular level remains obscure. Synaptic dysregulation of epileptic neuronal groups with unbalanced inter-relationship of excitatory and inhibitory complements within the epileptic focus has been only partly elucidated. Therefore, the treatment of epilepsy remains fortuitous to a great extent because idopathic epileptogenesis is stochastically unpredictable. We do not yet know why, how and when the epileptic process begins, and vice versa, why the process may occasionally be completely silent in a continuously overstrained interaction system. What link in the excitation neuronal system is weakest: membrane instability, neurotransmitter incoordination, or something else? Yet, in two thirds of the individuals prone to epilepsy, monotherapy with so-called channel antiepileptics with selective action on membrane ion exchange and/or antiepileptics with neurotransmitter modulation will stop the manifestation of epilepsy but not the longterm proneness to epileptogenesis. In the remaining one third of sufferers the epilepsy is refractory to any therapeutic attempts. Therefore, antiepileptic treatment is a highly complex, individualized procedure in which only the observational predictors have been positively defined to date, i.e. clinical manifestation and electroencephalography specificity. They make the basis for balanced evaluation of the real extent of action of the antiepileptics available. PMID- 15813354 TI - [Epilepsy--a review of its development in the world and in Croatia]. AB - Development of studies in epilepsy is presented, primarily in Croatia and briefly with regard to the world. The first epileptology available to the author were those published in Liecnicki viestnik in 1887 by D. Forenbacher (1896), by J. Tyrman, and in 1900 by F. Gutschy. The names Julius, Lopasic, Jelasic appeared from 1937 to 1943. In 1953, initiated by Lopasic in cooperation with Julius and Glaser, the journal Neuropsihijatrija was founded, dealing with neurology, psychiatry and peripheral fields. Thus began a new era in publishing in the field of epilepsy. The first practice for epilepsy was set up in 1952 by Dogan and Hajnsek, assisted by Lopasic, when the first EEG device (Medcraft) became available through UNRRA. By 1956 it expanded into a Clinic for Epilepsy, led by Hajnsek, soon becoming a meeting place for all those interested in epilepsy in children and adults. After two wars of work, the first locally designed EEG devices, produced by Santic and Kolaj, were supplied to six laboratories in 1961 throughout Croatia. In 1962 a section for EEG and Clinical Neurophysiology was founded at the Medical Association of Croatia. That year the Clinic for Epilepsy became the Centre for Epilepsy. In 1964 Croatian Section for EEG and Clinical Neurophysiology was founded in Ohrid, which became very active, organizing a number of meetings and ten symposia throughout former Yugoslavia. In 1972, the Croatian League for providing help to those suffering from epilepsy was founded, functioning independently, and organizing meetings, symposia and participating in the education of what by then had become a significant number of epileptologists treating children and adults, indirectly participating in the organization of more than seventy EEG laboratories throughout Croatia. During the 1990s, the Croatian League against Epilepsy also gained its independence and, with the Croatian Society for Epilepsy, in 1998 became a member of the ILAE and IBE (International Federation League against Epilepsy and International Bureau for Epilepsy). Both organizations are involved in the global campaign against epilepsy, entitled "Out of the Shadows", organizing meetings on the classification of epilepsy, diagnosis, and use of new anti-epileptics alongside those already existing. By giving lectures in kindergartens, schools, on radio and television, they are providing assistance to those suffering from the disease, helping them deal with their socioeconomic problems, in schooling, employment, marriage, having children, appearing before courts, handling themselves in road traffic, and in everyday life. PMID- 15813355 TI - [Minimally invasive neurosurgical methods in treatment of pharmacoresistant epilepsy]. AB - The first clinical attempts at neuromodulation in the form of applying functional electrostimulations started some thirty years ago. Nowadays, it is obvious that the approach to neuromodulation and functional electrostimulation has changed significantly. Neuromodulation tends to affect the disturbed function either by the modulation of neuronal signals or by abolition of dysfunction, preserving the intact central nervous system. The mechanism of activity is connected through direct afferent projections, neurotransmitter modulation and neuronal network regulation. NeuroCybernetic Prosthesis (NCP; Cyberonics) is a vagal nerve stimulator consisting of a pulse generator, bipolar VNS lead, programming wand with accompanying software for IBM-compatible computer, a tunneling tool and handheld magnets. NCP is placed on the left vagal nerve (middle cervical part). In 1988, Penry JK et al. inserted the first human implant. The Food and Drug Association indicated VNS as add-on therapy for diminishing the number of seizures in the adults and adolescents over 12 years of age with partial seizures, who are resistant to pharmacological therapy. PMID- 15813356 TI - [Epilepsy and pregnancy: antiepileptic drugs effects on pregnancy]. AB - PURPOSE: The aim of our study was to analyse the antiepileptic drugs (AEDs) usage in pregnant women with epilepsy, and to evaluate possible complications of this therapy during pregnancy and delivery as well as fetal effects of there drugs. METHODS: This retrospective study was conducted in the University Hospital Center Rijeka, Croatia, during the ten-year period (Jan 1, 1993-Dec 31, 2002). The study included 151 pregnant women with epilepsy. Data were entered in a structured form for evaluation of patient general data and data on antiepileptic therapy and neonatal outcome according to the use of AEDs. RESULTS: During the study period, 185 children (male 95) were born. Twenty-four miscarriages were recorded in women with the diagnosis of epilepsy in their medical history. In 117 of women, the onset of epilepsy occurred before the age of 20 years. AEDs were used in 111 of patients, in 77% of them as monotherapy. The most frequently used AEDs were carbamazepine (36.2%), valproate (29.3%), phenobarbitone (24.1%). Folic acid was used in 25 pregnancies only, however, during 2002 folic acid was used in 15 pregnancies (yielding 22 in total). Apgar score was equal or higher than 8/8 in 96% of children. No specific teratogenic effects were observed. CONCLUSION: Study results showed standard AEDs to be most commonly used in the study population, entailing no major birth defects, thus the potential risk probably being rather low. In the study population of pregnant women with epilepsy, the use of folic acid improved in 2002 in comparison to the previous period. PMID- 15813357 TI - [The risk of second seizure in children with benign childhood epilepsy with centrotemporal spikes without treatment--a prospective study]. AB - PURPOSE: To determine the rate of second seizure occurrence within and after six months of the first seizure in children with benign childhood epilepsy with centrotemporal spikes (BECTS) who did not undergo treatment after the first seizure. The results of this analysis may help elucidate the dilemma whether or not to treat the child after the first seizure. PATIENTS AND METHODS: Thirty-nine children with BECTS from our department (aged 3-11 years) were analyzed as candidates to be enrolled in a prospective multicenter randomized double-blind placebo controlled study on therapeutic efficacy of sulthiame. Thirty-four of 39 children were not treated after the first seizure. Four children were lost from the study, thus 30 children were included in final analysis. After the first seizure, the parents were instructed to apply diazepam rectal solution in case of second seizure, and were warned to observe the child, particularly during the first sleep and before awaking. RESULTS: Twenty of 30 (66.6%) children experienced second seizure within six months of the first one. Some of these children entered the group treated with sulthiame vs. placebo, and those who did not meet the criteria for sulthiame group were treated with carbamazepine. Ten of 30 (33.4%) children did not experience second seizure within six months of the first one. In only one of them, the second seizure occurred 14 months of the first one. The epileptic status did not appear as second seizure, irrespective of whether or not the children received rectal diazepam at seizure onset. CONCLUSIONS: In children with BECTS, a high incidence of second seizure was recorded within six months of the first seizure, whereas the rate of second seizure after six months of the first one was very low. The probability of the occurrence of epileptic status in children with BECTS could be neglected. These results may be viewed as a small contribution to clarifying the dilemma of whether or not, and when to treat children with BECTS. Because of the high incidence of second seizure, we decided to treat all children with BECTS after the first seizure. PMID- 15813358 TI - [Etiology and treatment of epilepsy in the elderly]. AB - Epilepsy currently affects 1.0%-1.5% of elderly population, and its prevalence and incidence rise as the number of old people increases worldwide. Antiepileptic drugs are the mainstay of treatment. Their phrarmacokinetics differ in old age and polypharmacotherapy often leads to drug interactions. Many of the affected have concomitant cardiovascular, metabolic, cerebrovascular, neurodegenerative or neoplastic diseases. Antiepileptic drugs add to the burden of medication and may produce neurotoxicity or cognitive impairment. PURPOSE: To give an overview of epidemiologic data and pharmacological management of epilepsy in the elderly. It is a common and important clinical problem. The increased awareness of this phenomenon has led to better understanding of the etiology of seizures and complexity of the pharmacokinetics in the elderly. METHOD: Data of 246 patients with epilepsy admitted to our Department during the year 2001 were retrospectively analyzed. RESULTS: Forty-three (17.4%) patients were older than 65. Thirty-two (74.4%) patients, 20 male and 12 female, had a newly diagnosed epilepsy. The etiology of seizures included cerebrovascular diseases in 22 (68.8%), trauma in 4 (12.5%), primary tumors in 3 (9.4%) and metastases in 3 patients (9.4%). The initial antiepileptic drug (AED) was phenytoin in 15 (46.9%), carbamazepine in 14 (43.8%), carbamazepine plus phenytoin in 2 (6.3%), valproic acid in 1 (3.1%), diazepam in 3 (9.4%) and oxazepam in 2 (6.3%) patients. Later, during follow-up, lamotrigine was introduced in 4 (12.5%) patients as a monotherapy and in 3 (9.4%) patients as add-on therapy. Topiramate was introduced in 1 patient as monotherapy and add-on therapy each. Gabapentin was introduced in one patient as add-on therapy. CONCLUSION: Cerebrovascular disease is the most common cause of new-onset seizures in the elderly. Pharmacological treatment is complicated by the age related changes of pharmacokinetics and pharmacodynamics and drug-drug interactions. Phenytoin and sodium valproate are the first choice agents for generalized tonic-clonic seizures, with carbamazepine preferred for partial seizures. The newer AEDs, such as gabapentin and lamotrigine, also warrant some consideration as the first-line agents because of their efficacy and favorable effect profile. The optimal management of epilepsy requires rapid investigation, accurate diagnosis, effective therapy, education and assured support. PMID- 15813359 TI - [Electroencephalography in status epilepticus in sleep (ESES) in various clinical pictures]. AB - Electroencephalographic epileptic status during sleep (ESES--according to Electrical Status Epilepticus in Sleep) is characterized with paroxysmal attacks of 1.5-3.5 (even to 5) Hz/s spikes and waves during NREM. A case is presented of a 7-year-old boy who had normal development until 3 years of age when epileptic attacks started. First to appear were left-sided, hemifacial twitches with orofacio-lingual deficits. Despite treatment with various types and combinations of antiepileptic medications, the attacks persisted and became more frequent. About a year after the onset of the disease, the spectrum of epileptic attacks had expanded (left-sided tonic-clonic, atonic-astatic, myoclonic, atypical absences, and then drop attacks and negative myoclonic seizures became dominant). The boy appeared mentally retarded. During the course of the disease, the diagnoses varied: hemifacial twitches, partial epilepsy, left-sided partial epilepsy, atypical benign epilepsy, Landau-Kleffner syndrome, myoclonic-astatic epilepsy. A year and a half after the onset of the disease EEG (polysomnographic whole-night recording) revealed electrical status epilepticus in sleep (ESES) with spike-wave index > 85%. It appeared that this was continuous spike and wave during slow wave sleep syndrome (CSWS) with characteristic attacks, bifrontal atrophies on brain CT and right frontotemporal EEG focus. Treatment with valproate and ethosuximide combined with vigabatrin and clonazepam seemed to be effective. In the last 20 months ESES is rare and transitory, mental and neuropsychological functions have improved, but he still has several hemifacial twitches daily. The two brain MRI scans were normal. Differential diagnosis can be atypical benign epilepsy (suspected familial diagnosis). Family history of atopy, 'persistent' colds and obstructive bronchitis in the boy and treatment with antiepileptic medications (especially carbamazepine, phenobarbital and phenytoin) could have been provoking or modulating factors of the epileptic disease. So far we have not identified an epileptic syndrome because it seems that multiple clinical pictures overlap. ESES could be diagnosed using polysomnographic whole-night recording with calculated spike-wave index. Follow up of the spike-wave index could be useful in differentiation, but not in precise diagnosis of epileptic syndrome. Clinical course could be modulated by different factors. Continuous following over a long period could be helpful in the classification of epilepsy. PMID- 15813360 TI - [Guidelines for management of epilepsy--commentary on Scottish ("SIGN") guidelines]. AB - The choice of AED (antiepileptic drug), worldwide and in Croatia, is been still based on the physician's subjective decision, personal experience, knowledge and marketing pressure made by big pharmaceutical industries. Therefore, according to some opinions, there is a need of treatment guidelines for epilepsy that would provide relevant information based on scientific evidence on the efficacy, tolerability and safety of AEDs. The guidelines, published by a competent source, should be designed as to allow for easy access to the information on the best practice in specific cases. An extensive background literature review was made to identify such a type of guidelines for the management of epilepsy. The literature review revealed a number of references with the recommendations for treating epilepsy in different groups of patients and from various, specific aspects of epilepsy treatment. However, only one comprehensive set of guidelines for the diagnosis and treatment of epilepsy treatment was found, i.e. the evidence-based guidelines published by the Scottish Intercollegiate Guidelines Network (SIGN). The development of this set of guidelines is quite extensively described in order to illustrate how rigorous and long-lasting the process was, including a great number of health professionals at the national level. Such a type of well designed guidelines facilitates access to highest educational standards for all professionals involved in the primary and secondary care of people with epilepsy. However, it is clear that guidelines can fully replace the standards of clinical practice based on critical evaluation and integration of all clinical data of each individual patient. No guidelines can replace the physician's obligation to keep informed of the novel achievements in the epileptology either. PMID- 15813361 TI - Exudate flavonoids of eight species of Ceanothus (Rhamnaceae). AB - Leaf glands of Ceanothus species excrete a lipophilic material that contains a variety of flavonoids. Most of these are aglycones, but some glycosides were also observed. Seven out of eight species exhibit flavonols, whereas flavones are excreted by only one species. Four species produce flavanones and dihydroflavonols; one excretes a remarkable quantity of flavonol glycosides. The exudate flavonoids thus form different patterns that might be characteristic for different Ceanothus species. PMID- 15813362 TI - Comparison of the volatile composition of Stachys persica Gmel. and Stachys byzantina C. Koch. oils obtained by hydrodistillation and steam distillation. AB - The oils obtained by hydrodistillation and steam distillation of the aerial parts of Stachys persica Gmel. and Stachys byzantina C. Koch grown in Iran were analyzed by GC/MS. The essential oil obtained by hydrodistillation of the aerial parts of S. persica was characterized by a high amount of non-terpenoid components of which methyllinoleate (27.7%), hexadecanoic acid (9.8%) and 6,10,14 trimethyl-2-pentadecanone (9.2%) were the major constituents, whereas the steam distilled oil of the plant contained hexadecanoic acid (27.2%), carvacrol (9.4%) and eugenol (5.2%). Both hydrodistilled and steam distilled essential oils of the aerial parts of S. byzantina were rich in sesquiterpenes such as a-copaene (16.6% and 10.4%), spathulenol (16.1% and 18.5%) and beta-caryophyllene (14.3% and 13.5%), respectively. PMID- 15813363 TI - Antioxidant and immunomodulatory constituents of henna leaves. AB - The immunomodulatory bioassay-guided fractionation of the methanolic extract of henna (Lawsonia inermis L.; syn. Lawsonia alba L.) leaves resulted in the isolation of seven compounds; three have been isolated for the first time from the genus, namely p-coumaric acid, 2-methoxy-3-methyl-1,4-naphthoquinone and apiin, along with the previously isolated compounds: lawsone, apigenin, luteolin, and cosmosiin. Structural elucidation of the isolated compounds was based upon their physical, chemical as well as spectroscopic characters. Their immuomodulatory profile was studied using an in vitro immunoassay, the lymphocyte transformation assay. The ABTS [2,2'-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid)], free radical scavenging assay depicted that all isolated compounds exhibited antioxidant activity comparable to that of ascorbic acid. PMID- 15813364 TI - Antioxidant metabolites from Limonium brasiliense (Boiss.) Kuntze. AB - A free radical scavenging activity guided fractionation of the polar extract from roots of Limonium brasiliense (Plumbaginaceae) led to the isolation of five active compounds including: myricetin 3-O-alpha-rhamnopyranoside (1), (-) epigallocatechin 3-O-gallate (2), (-)-epigallocatechin (3), (+)-gallocatechin (4) and gallic acid (5). These and other chemical constituents are reported for the first time for this species. The characterization of these compounds was achieved by spectroscopic methods (1H NMR, 13C NMR and UV). PMID- 15813365 TI - Antioxidant activities of three dihydrochalcone glucosides from leaves of Lithocarpus pachyphyllus. AB - In vitro antioxidant activities of three sweet dihydrochalcone glucosides from the leaves of Lithocarpus pachyphyllus (Kurz) Rehd. (Fagaceae), trilobatin 2" acetate (1), phloridzin (2) and trilobatin (3), were investigated. The IC50 (50% inhibitory concentration) values for compounds 1-3 of lipid peroxidation in rat liver homogenate were 261, 28, 88 microM, respectively. Compounds 1-3 increased superoxide dismutase (SOD) activity with EC50 (50% effective concentration) values of 575, 167, 128 microM, and glutathione peroxidase (GSH-Px) activity with EC50 values of 717, 347, 129 microM, respectively, and showed only weak DPPH (1,1 diphenyl-2-picrylhydrazyl) radical scavenging activity. PMID- 15813366 TI - Immunomodulating polysaccharide fractions of Menyanthes trifoliata L. AB - Looking for new plant sources of immunomodulating agents polysaccharide-rich fractions (PS) from Menyanthes trifoliata L. (Menyanthaceae) have been isolated. The herb of Menyanthes trifoliata L. was sequentially extracted with water, 0.1 M NaOH, 8% CH3COOH, and 1 M NaOH. After dialysis and resolution on Biogel P-10 four homogenic (B-4, B-5, C-4, D-5) and two nonhomogenic (A-3 and D-4) PS were isolated. About 0.5% of PS over 3500 Da were found in the dry plant material. They were characterized through chemical analysis, NMR and vibrational spectroscopy. Speciation analysis of chosen metal/metaloid elements was performed and an exceptionally high concentration of Se was found in PS of a pure water extract (A-3). The biological tests on the immunomodulating influence with human blood-derived lymphocytes and granulocytes revealed that two fractions, B-4 and B 5, were strong stimulators of immune cells, whereas fractions D-5 and A-3 were found as potent suppressive and anti-inflammatory agents. The applied isolation procedures led to the separation of active compounds into stimulatory and inhibitory fractions. PMID- 15813367 TI - Bioactivity of Lignans from Taxus baccata. AB - Three lignan derivatives, (-)-taxiresinol (1), (-)-3'-demethylisolariciresinol-9' hydroxyisopropylether (2) and (-)-3-demethylisolariciresinol (3), previously isolated from the heart-wood of Taxus baccata L. (Taxaceae) were investigated for cytotoxicity against the Oncology Cell Line Panel (breast, colon, ovary, prostate, lung and a normal adult bovine aortic endothelial cell line) as well as for antimicrobial activities. Besides, a chloroform-soluble portion of the ethanol extract from the heartwood of T. baccata was also tested for antimicrobial activities. Compounds 1 - 3 did not demonstrate much cytotoxic potency according to the reference drug etoposide. With the exception of compound 2, the other two lignans (1 and 3) and the chloroform extract were shown to possess antifungal activity, whereas only the chloroform extract exhibited antibacterial activity against Pseudomonas aeruginosa. PMID- 15813368 TI - A new heptasubstituted (E)-aurone glucoside and other aromatic compounds of Gomphrena agrestis with biological activity. AB - A new aurone 1 and two known substances, aurantiamide acetate (2) and tiliroside (3), were isolated from the ethanolic extract of Gomphrena agrestis. The structural determination of 1 was based on spectroscopic and spectrometric data. The substance was defined as (E)-3'-O-beta-D-glucopyranosyl-4,5,6,4'-tetrahydroxy 7,2'-dimethoxyaurone. Biological activity of the ethanolic crude extract and isolated compounds against bacteria, fungi and Leishmania amazonensis amastigotes was evaluated. This appears to be the first report documenting aurone and aurantiamide compounds in the Amaranthaceae family. In the evaluation of biological activity the ethanolic extract of G. agrestis and compounds 1, 2, and 3 were shown to be active mainly against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa. PMID- 15813369 TI - Umbelliprenin from Ferula persica roots inhibits the red pigment production in Serratia marcescens. AB - The chloroform extract of Ferula persica var. persica roots was found to inhibit red pigment production of Serratia marcescens. A bioguided fractionation study by preparative thin layer chromatography (PTLC) detected a fraction (Rf = 0.71, petroleum ether/EtOAc, 2:1 v/v), which was effective on depigmentation of Serratia marcescens. Using conventional spectroscopy methods, the active fraction was identified as umbelliprenin. Neither the chloroform extract nor the isolated umbelliprenin fraction showed any antibacterial activity against the test strain at a certain concentration. In contrast, they exhibited depigmentation zones on culture plates. PMID- 15813370 TI - Phenolic compounds and flavonoids as plant growth regulators from fruit and leaf of Vitex rotundifolia. AB - Five phenolic compounds, 4-hydroxybenzoic acid methyl ester (1), vanillic acid methyl ester (2), 4-hydroxy benzaldehyde (3), 4-hydroxybenzoic acid (4) and ferulic acid (5), and four flavonoids, 5,5'-dihydroxy-4',6,7-trimethoxyflavanone (6), luteolin (7), vitexicarpin (8) and artemetin (9), were isolated from fruits and leaves of Vitex rotundifolia L. The biological activities of these nine compounds have been examined using a bioassay with lettuce seedlings. PMID- 15813371 TI - Binary mixtures of (N-phosphonomethyl)-glycine with new aminophosphonates. AB - The potential biological activity of binary mixtures of some new organophosphorous compounds, aminoalkane- and aminofluorenephosphonates, with (N phosphonomethyl)-glycine (glyphosate, PMG) was studied. The inhibition of growth of wheat (Triticum aestivum) induced by individual compounds and their equimolar mixtures with PMG was a measure of that activity. The experiments were expected to show if the new compounds exhibited good biological activity to be used for agrochemical applications and if this activity can be improved when they are used in mixtures with glyphosate which is the active component of the well-known herbicide Roundup. The results obtained show that aminofluorenephosphonates inhibited wheat growth when used in micromolar concentrations. Thus, their efficiency can be compared to that of PMG. The efficiency of aminoalkanephosphonates was one order of magnitude weaker. The measure of the efficiency was the effective concentration inhibiting wheat growth by 50% (EC50). The most demanded interaction, i.e., a synergistic was observed for only one of binary mixtures of the compounds studied with PMG. Mostly they showed antagonistic or strong antagonistic interactions. Some of them were of the additive type. Such results exclude the possibility of potential use of all the compounds studied in binary mixtures with phosphonomethylglycine, especially as the mentioned synergistic interaction found was rather weak. The influence of structural features of anminophosphonates on the results obtained is discussed. PMID- 15813372 TI - Cytotoxic activity of pisosterol, a triterpene isolated from Pisolithus tinctorius (Mich.: Pers.) Coker & Couch, 1928. AB - Pisolithus tinctorius (Basidiomycete) is an ectomicorrhizal fungus found in the roots and soil surrounding of many species of eucalyptus and pine trees. The present work verified the cytotoxic potential of pisosterol, a triterpene isolated from P. tinctorius collected in the Northeast region of Brazil, on three different animal cell models: mouse erythrocytes, sea urchin embryos and tumor cells. Pisosterol lacked activity on mouse erythrocytes as well as on the development of sea urchin eggs, but strongly inhibited the growth of all seven tumor cell lines tested, especially the leukemia and melanoma cells (IC50 of 1.55, 1.84 and 1.65 microg/ ml for CEM, HL-60 and B16, respectively). The results found for pisosterol were compared with those of doxorubicin and etoposide. PMID- 15813373 TI - Antiproliferative effect of Angelica archangelica fruits. AB - The aim of this work was to study the antiproliferative effect of a tincture from fruits of Angelica archangelica and the active components using the human pancreas cancer cell line PANC-1 as a model. Significant dose-dependent antiproliferative activity was observed in the tincture with an EC50 value of 28.6 microg/ml. Strong antiproliferative activity resulted from the two most abundant furanocoumarins in the tincture, imperatorin and xanthotoxin. The contribution of terpenes to this activity was insignificant. Imperatorin and xanthotoxin proved to be highly antiproliferative, with EC50 values of 2.7 microg/ml and 3.7 microg/ml, respectively, equivalent to 10 and 17 microM. The results indicate that furanocoumarins account for most of the antiproliferative activity of the tincture. PMID- 15813374 TI - Complement inhibiting properties of dragon's blood from Croton draco. AB - The latex of Croton draco, its extracts and several latex components have been investigated for their influence on both classical (CP) and alternative (AP) activation pathways of the complement system using a hemolytic assay. The best inhibition was found for the classical pathway. The latex, ethyl acetate and ethyl ether extracts exhibited extremely high inhibition on the CP (94, 90 and 77%, respectively) at a concentration of 1 mg/ml. The flavonoid myricitrin, the alkaloid taspine and the cyclopeptides P1 and P2 showed high inhibition on CP (83, 91, 78 and 63%, respectively) at a concentration of 0.9 mM. PMID- 15813375 TI - Analysis of Rhioxma Curcumae Aeruginosae volatiles by solid-phase microextraction with gas chromatography-mass spectrometry. AB - In this paper, a headspace solid-phase microextraction (HS-SPME) method was applied to analyse the volatile compounds in a traditional Chinese medicine (TCM), Rhioxma Curcumae Aeruginosae. SPME parameters such as fibers, extraction temperature, extraction time and desorption time were investigated. Thirty-five volatile compounds were separated and identified. Relative standard deviations (RSDs) were less than 8.4%, showing that the method has a good reproducibility. The volatile constituents were also analyzed by steam distillation (SD) and thirty-seven compounds were identified. The similar results obtained by the two methods showed that SPME is a good alternative for the analysis of volatile constituents in Rhioxma Curcumae Aeruginosae samples and it is a relatively simple, rapid and solvent-free method. PMID- 15813376 TI - Green paramecia as an evolutionary winner of oxidative symbiosis: a hypothesis and supportive data. AB - A single cell of the green paramecia (Paramecium bursaria) harbors several hundreds of endo-symbiotic Chlorella-like algae in its cytoplasm. Removal of algae from the host organism and re-association of ex-symbiotic host paramecia with ex-symbiotic algae can be experimentally demonstrated in the laboratory. However, the mechanism precisely governing the alga-protozoan association is not fully understood, and the origin of symbiosis in the evolutionary view has not been given. Here, we propose the possible biochemical models (models 1 and 2) explaining the co-evolution between Paramecium species and algal symbionts by pointing out that algal photosynthesis in the host paramecia plays a dual role providing the energy source and the risk of oxidative damage to the host. Model 1 lays stress on the correlation between the (re)greening ability of the paramecia and the tolerance to oxidative stress whereas model 2 emphasizes the cause of evolutionary selection leading to the emergence of Paramecium species tolerant against reactive oxygen species. PMID- 15813377 TI - Quantitative changes of secondary metabolites of Matricaria chamomilla by abiotic stress. AB - The responses of young plants of diploid and tetraploid Matricaria chamomilla cultivars to abiotic stress were studied. The course of quantitative changes of main leaf secondary metabolites was evaluated within an interval from 6 h before to 54 h after spraying the leaf rosettes with aqueous CuCl2 solution. The content of herniarin in the treated plants rose approximately 3 times, simultaneously with a decline of its precursor (Z)- and (E)-2-beta-D-glucopyranosyloxy-4 methoxycinnamic acid. The highest amounts of umbelliferone in stressed plants exceeded 9 times and 20 times those observed in control plants of the tetraploid and diploid cultivar, respectively. Due to stress the concentration of ene-yne dicycloether in leaves decreased by more than 40%. The pattern of quantity changes of the examined compounds in tetraploid and diploid plants was similar. PMID- 15813378 TI - The very-long-chain fatty acid synthase is inhibited by chloroacetamides. AB - The first elongation step to form very-long-chain fatty acids (VLCFAs) is catalyzed by the VLCFA-synthase. CoA-activated fatty acids react with malonyl-CoA to condense a C2-unit. As shown with recombinant enzyme this reaction is specifically inhibited by chloroacetamide herbicides. The inhibition is alleviated when the inhibitor (e.g. metazachlor) is incubated together with adequate concentrations of the substrate (e.g. oleoyl-CoA). Malonyl-CoA has no influence. However, once a chloroacetamide has been tightly bound to the synthase after an appropriate time it cannot be displaced anymore by the substrate. In contrast, oleoyl-CoA, is easily removed from the synthase by metazachlor. The irreversible binding of the chloroacetamides and their competition with the substrate explains the very low half-inhibition values of 10(-8) M and below. Chiral chloroacetamides like metolachlor or dimethenamid give identical results. However, only the (S)-enantiomers are active. PMID- 15813379 TI - Rosmarinic acid synthesis in transformed callus culture of Coleus blumei benth. AB - Agrobacteria mediated Coleus blumei tumour tissues were cultured in vitro on MS medium. Sixteen diversified transformed callus cultures were maintained for several years in the absence of plant growth regulators and antibiotics without affecting the growth rate. Rosmarinic acid was detected spectrophotometrically in all tissue lines but in different quantities. The highest rosmarinic acid accumulation detected was 11% of dry tissue mass. The relation between culture growth and rosmarinic acid production was investigated in three callus lines. The lines showed different rosmarinic acid accumulation in relation to their growth rate; it was either parallel or inversely related to the tissue growth. The effects of certain medium constituents on the callus growth and rosmarinic acid accumulation were examined in four tumour cell lines. Addition of 4% or 5% sucrose stimulated rosmarinic acid synthesis and decreased callus growth. Nitrogen reduction to one half or one quarter of initial concentration did not affect rosmarinic acid synthesis and decreased callus growth in three lines, while it increased rosmarinic acid accumulation and callus growth in one line. Addition of 0.1 mg/l Phe stimulated rosmarinic acid production in two lines but had little effect on the rosmarinic acid level in others. Rosmarinic acid production was significantly improved on modified macronutrients, where the Ac2 line produced 16.5 mg of rosmarinic acid per tube (0.2 g of dry wt) after being in culture for 35 days. PMID- 15813380 TI - Cloning and sequencing of hydroxylase genes involved in taxol biosynthesis. AB - Two full-length cDNAs (TCH1 and TCH2) were obtained from a cDNA library of Taxus chinensis mainly by the single specific-primer PCR (SSP-PCR) method. Compared with other reported enzymes from Taxus species, the deduced amino acid sequences of TCH1 and TCH2 exhibit significant homologies to hydroxylases that are involved in taxol biosynthesis. These findings imply that the two new genes are closely related to the biosynthesis of taxol/taxoids. Data Accession No: AF545833 and AY374652. PMID- 15813381 TI - Proteins with spectrin motifs which do not belong to the spectrin-alpha-actinin dystrophin family. AB - Using several consensus sequences for the 106 amino acid residue alpha-spectrin repeat segment as probes we searched animal sequence databases using the BLAST program in order to find proteins revealing limited, but significant similarity to spectrin. Among many spectrins and proteins from the spectrin-alpha-actinin dystrophin family as well as sequences showing a rather high degree of similarity in very short stretches, we found seven homologous animal sequences of low overall similarity to spectrin but showing the presence of one or more spectrin repeat motifs. The homology relationship of these sequences to alpha-spectrin was further analysed using the SEMIHOM program. Depending on the probe, these segments showed the presence of 6 to 26 identical amino acid residues and a variable number of semihomologous residues. Moreover, we found six protein sequences, which contained a sequence fragment sharing the SH3 (sarc homology region 3) domain homology of 42-59% similarity. Our data indicate the occurrence of motifs of significant homology to alpha-spectrin repeat segments among animal proteins, which are not classical members of the spectrin-alpha-actinin dystrophin family. This might indicate that these segments together with the SH3 domain motif are conserved in proteins which possibly at the early stage of evolution were close cognates of spectrin-alpha-actinin-dystrophin progenitors but then evolved separately. PMID- 15813382 TI - Genomic relations among two non-mangrove and nine mangrove species of Indian Rhizophoraceae. AB - Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers were used to study the genomic relationship among 11 members of Indian Rhizophoraceae represented by nine true mangroves and two non mangrove species. The AFLP and RAPD bands were scored and analyzed for genetic similarities and cluster analysis was done which separated the 11 species studied into two main groups, the true mangroves and the non-mangroves. The polymorphism observed for these markers showed a high degree of genetic diversity among the constituent taxa of the family. The phylogenetic relationship inferred from molecular marker systems supported the traditional taxonomic classification of the family Rhizophoraceae based on morphological characters at the levels of tribe, phylogeny and delimitation of genera and species, except the intra-generic classification of the genus Bruguiera and the placement of Rhizophora in the family Rhizophoraceae. PMID- 15813383 TI - New products of defense secretion in south east Asian whip scorpions (Arachnida: Uropygi: Thelyphonida). AB - Secretion products from the opisthosomal defense gland of south east Asian whip scorpions were identified for the first time by gas-chromatography and mass spectrometry. Specimens of the genera Hypoctonus, Typopeltis and Ginosigma were tested. While some ingredients are present in large concentrations, others are possibly only side products and may be synthesized more incidentally. For this reason no important functional role is attributed to them. There are considerable individual differences concerning the concentrations of various ingredients. While the secretion products of most species of the genus Typopeltis--similar to Mastigoproctus--are characterized by acetic and octanoic acid in large concentrations, the secretion product of Hypoctonus siamensis provides octanoic acid only in a very low concentration but it is characterized by hexyl acetate. PMID- 15813384 TI - Exocrine gland secretions of virgin queens of five bumblebee species (Hymenoptera: Apidae, Bombini). AB - Secretions of three different glands (mandibular gland, labial gland, and Dufour's gland) of virgin queens of five bumblebee species (Bombus lucorum, B. lapidarius, B. hypnorum, B. pascuorum, and B. terrestris) were analysed. Around 200 compounds were identified in the secretions. The compositions of the secretions of labial and mandibular glands were species-specific. Dufour's gland of all species produced mainly hydrocarbons, both saturated and unsaturated, the proportions of which differed quantitatively between the species studied. PMID- 15813385 TI - A serine protease from a detergent-soluble extract of Leishmania (Leishmania) amazonensis. AB - Proteases mediate important crucial functions in parasitic diseases, and their characterization contributes to the understanding of host-parasite interaction. A serine protease was purified about 43-fold with a total recovery of 60% from a detergent-soluble extract of promastigotes of Leishmania amazonensis. The purification procedures included aprotinin-agarose affinity chromatography and gel filtration high performance liquid chromatography. The molecular mass of active enzyme was 110 kDa by native gel filtration HPLC and by SDS-PAGE gelatin under non-reducing conditions. Under conditions of reduction using SDS-PAGE gelatin analyses the activity of enzyme was observed in two proteins of 60 and 45 kDa, suggesting that the enzyme may be considered as a dimer. The Leishmania protease was not glycosylated, and its isoelectric point (pI) was around 4.8. The maximal protease activity was at pH 7.0 and 28 degrees C, using a-N-o-tosyl-L arginyl-methyl ester (L-TAME) as substrate. Assays of thermal stability indicated that this enzyme was totally denatured after pre-treatment at 42 degrees C for 12 min and preserved only 20% of its activity after pre-treatment at 37 degrees C for 24 h, in the absence of substrate. Hemoglobin, bovine serum albumin (BSA), ovalbumin and gelatin were hydrolyzed by Leishmania protease. Inhibition studies indicated that the enzyme belonged to a serine protease class because of a significant impediment by serine protease inhibitors such as benzamidine, aprotinin, and antipain. The activity of the present serine protease is negatively modulated by calcium and zinc and positively modulated by manganese ions. This is the first study that reports the purification of a protease from a detergent-soluble extract of Leishmania species. PMID- 15813386 TI - Inhibition of cytochrome P450 mediated enzyme activity by alkylphosphocholines. AB - The inhibitory potency of four alkylphospholipids: rac-1-O-phosphocholine-2 hydroxy-octadecane (rac-2-OH), rac-1-O-phosphocholine-2-O-acetyl-octadecane (rac 2-O-acetyl), rac-1-O-phosphocholine-2-amino-octadecane (rac-2-NH2) and rac-1-O phosphocholine-2-N-acetyloctadecane (rac-2-N-acetyl), on the cytochrome P450 dependent monooxygenase activity has been evaluated. The IC50 values of the alkylphosphocholines with 7-ethoxycoumarin as substrate in liver microsomal fractions of PB-treated rats and with a reconstituted CYP2B1: NADPH-P450 reductase system are in the range of 3.2-5.0 microM and 2.8-3.5 microM, respectively. Lineweaver-Burk plots with the inhibitors in concentrations that were found to cause roughly a 50% inhibition and with 7-ethoxycoumarin as substrate revealed for all four alkylphospholipids a competitive inhibition type. The degree of the competitive inhibition is quantified by the Ki values. With liver microsomal fractions of PB-treated rats, the Ki values of rac-2-OH (Ki = 1.36 microM) and rac-2-O-acetyl (Ki = 1.33 microM) differs slightly from those of rac-2-NH2 (Ki = 2.2 microM) and rac-2-N-acetyl (Ki = 2.2 microM), but with the reconstituted CYP2B1: NADPH-P450-reductase system all Ki values are in the small range of 1.8 - 2.6 microM, indicating that the short substituted group at the 2 position (OH; O-acetyl; NH2; N-acetyl) of the long chain octadecanol part of the phosphodiesters exhibit no essential role on the strong inhibitory potency of these alkylphosphocholines on the 7-ethoxycoumarin-O-deethylase activity. PMID- 15813387 TI - Thymol derivatives from a root culture of Inula helenium. AB - A root culture of Inula helenium L. was established from leaf explants of aseptic seedlings. An ethanol extract from the lyophilised roots was fractionated using different chromatographic techniques (CC, TLC). The main secondary metabolites found in the root culture were two thymol derivatives: 10-isobutyryloxy-8,9-epoxy thymol isobutyrate (1) and 10-isobutyryloxy-6-methoxy-8,9-epoxy-thymol isobutyrate (2). The compounds were identified by spectral methods. Quantification of compound 1 in plant material was done by analytical RP-HPLC. PMID- 15813389 TI - [Notes on diet during the time of Cervantes]. PMID- 15813388 TI - The antiproliferative agents trans-bis(resorcylaldoximato)copper(II) and trans bis(2,3,4-trihydroxybenzaldoximato)copper(II) and cytopathic effects of HIV. AB - trans-Bis(resorcylaldoximato)copper(II) and trans-bis-(2,3,4 trihydroxybenzaldoximato)copper(II) (CuRES2 and CuTRI2, respectively) have been tested for antiviral properties against HIV, using an in vitro assay that measures the ability of the test compounds to prevent the killing of susceptible human cells by HIV. In the case of CuTRI2, T4 lymphocytes (CEM-V and CEM-Z cell lines) were exposed to HIV at a virus to cell ratio approx. 0.05 in microtiter plates. In the case of CuRES2, a human leukemia cell line (MT-2) was used instead. The tetrazolium salt XTT was added to all wells, and the cultures were incubated and analyzed spectrophotometrically to quantitate formazan production and viewed microscopically for detection of viable cells. In spite of their antiproliferative properties, neither agent had any detectable ability to prevent the cytopathic effects of HIV in cultures of the target cells used. Because the test system employed was constructed in such a way as to detect antiviral agents acting at any stage of the virus reproductive cycle, the results obtained strongly suggest that neither studied agent has any value as the direct prevention of the cell destruction caused by HIV is concerned. PMID- 15813390 TI - Conclusions of the II SENPE discussion forum on: hospital malnutrition. PMID- 15813391 TI - [Nutrition in the end-stage patient. The ethical point of view]. PMID- 15813392 TI - [Early enteral nutrition in the critically-ill patient]. AB - Enteral nutrition has demonstrated to be a useful and safe method to nourish critically ill patients admitted to the Intensive Care Unit. Although the time a severely ill patient can stand without nutrition is unknown, accelerated catabolism and fasting may be deleterious in those patients, and the more common recommendation is to start on artificial nutrition when a fasting period longer than seven days is foreseen. At an experimental level, advantages of enteral nutrition over parenteral nutrition are evident since the use of nutritional substrates via the gastrointestinal tract improves the local and systemic immune response and maintains the barrier functions of the gut. Clinical studies have demonstrated that early enteral nutrition administered within the first 48 hours of admission decreases the incidence of nosocomial infections in these patients, but not the mortality, with the exception of special groups of patients, particularly surgical ones. The major inconvenience of enteral nutrition is its digestive intolerance and the transpyloric approach, necessary when there is gastroparesia. Its efficacy is also questioned when the patient has tissue ischemia. For early enteral nutrition to be effective, a treatment strategy must be implemented that includes from simple measures, such as uprising the bed headrest, to more sophisticated ones, such as the transpyloric approach or the use of nutrients with immunomodulatory capabilities. To date, the use of early enteral nutrition is the best method for nutritional support in this kind of patients provided that it is individualized according to each patient clinical status and that is done following an adequate therapeutic strategy. PMID- 15813393 TI - Food, energy and macronutrient intake of postmenopausal women from a menopause program. AB - OBJECTIVE: This study aimed to analyse the food, energy and macronutrient intake of a group of postmenopausal women participating in a health-care-program. METHODS: Subjects included were 38 healthy postmenopausal women aged between 46 and 60 years, recruited from the Menopause Program of the Madrid City Council. Physical activity, some anthropometric data and dietary information was obtained using a modified version of the dietary history method, which contained a 24-hour recall and a food frequency questionnaire covering the preceding month as reference period. Dietary quality indexes, including those of the energy provided by macronutrients, alcohol and fatty acids and PUFA + MUFA/SFA and PUFA/SFA ratios were calculated. RESULTS: This group consumed a diet very similar to the traditional Mediterranean diet. Intake of vegetables (415 +/- 165 g/d), fruits (396 +/- 178 g/d) and fish (131 +/- 69 g/d) was high and a wide variety of these products was consumed. Potato and cereal (157 +/- 76 g/d) intake was low. Dairy products, meat, poultry and eggs were only a modest part of this diet. The fat quality (PUFA + MUFA/SFA = 2.26) was satisfactory. CONCLUSION: The diet of this group of postmenopausal women attending a prevention program closely conforms to current nutritional guidelines. Physical activity, body weight and intake of vegetable foods are adequate and may be very useful to counterbalance the increased risk of several pathologies after menopause. However, consumption of carbohydrate rich foods is lower than recommended. Participation in the menopause health-care-program is useful for weight control and dietary advice. PMID- 15813394 TI - [Nutrients and energy intake assessment in the critically ill patient on enteral nutritional therapy]. AB - INTRODUCTION AND OBJECTIVES: The critically ill patient is especially susceptible to malnutrition due to his/her hypermetabolic state that leads to an increase in the nutritional requirementes, which many times are not compensated with the administered enteral formulas. The assessment of nutritional intake is essential in this kind of patients to know to what level their energetic and nutritional requirements are fulfilled, improving and monitoring in the most individualized possible way to indicated clinical and nutritional therapu. METHODOLOGY: This is a retrospective study in which all patients admitted to the Intensive Care Unit of Virgen de las Nieves Hospital were studied from January to December of 2003, aged more than 18 years, and on enteral nutrition. A total of 90 patients (52 men and 38 women) were studied, 81% of which were older than 50 years, and 57% had hospital stays longer than 8 days, with a 21% mortality rate. Intake was assessed from time of admission and throughout the whole hospitalization period. Energetic requirements were calculated according to the modified Long's formula and micronutrients intakes were compared to existing general recommendations for the Spanish, European and American populations, and to vitaminic requirements in critically ill patients. RESULTS: Percentages of mean energy and nutrients intakes in relation to theoretical calculated requirements for both genders are presented in figure 1. Mean energy intake was 1,326 cal in men and 917 cal in women. With regards to micronutrients intake, the values found for proteins, falts, and carbohydrates were lower than 50% of the requirements for both genders. The percentage of adequacy as referred to requirements for vitamins and minerals intake is shown in figure 2. Reference recommendations used correspond to sufficient intakes to cover the healthy individual requirements, therefore, the values obtained in our study show and adequacy greater than 75%, with the exception of particular elements such as vitamin A and magnesium. However, by taking a look at figure 3, which shows the adequacy of vitamins intake at recommended does for sick patients, the intake is lower than 25% of the requirements in all cases, and these deficiencies significantly interfere with wound healing, the immune, cardiovascular and nervous systems, as well as with metabolism of the remaining macronutrients leading to an unbalanced situation of the antioxidant system, worsening the patient's clinical status. CONCLUSIONS: The present study confirms the need for monitoring individually the nutritional requirements in the critically ill patient and adapting recommendations to his/her metabolic changes, since currently these recommendations are not clearly defined for these situations. It is necessary to provide micronutrients doses closer to the patient's demands, so that the nutritional status and the balance of the antioxidant system may be preserved or improved, making the adopted clinical treatment more effective. PMID- 15813395 TI - Iron metabolism, inflammation and anemia in critically ill patients. A cross sectional study. AB - INTRODUCTION: For critically patients, enteral immunonutrition results in notable reductions in infections and in length of stay in hospital, but not on mortality, raising the question as to whether this relate to the heterogeneous nature of critically ill patients or to the absence of the altered absorption of specific nutrients within the immunonutrient mix (e.g. iron). Immune-associated functional iron deficiency (FID) is not only one of the many causes or anaemia in the critically ill, but also a cause of inappropriate immune response, leading to a longer duration of episodes of systemic inflammatory response syndrome and poor outcome. OBJECTIVE: This prospective cross-sectional study was undertaken to assess the prevalence of FID in critically ill patients during their stay in intensive care (ICU) in order to find the more appropriate population of patients that can benefit from iron therapy. METHOD: Full blood cell counts, including reticulocytes (RETIC), serum iron (SI), transferring levels (TRF) and saturation (satTRF), serum TFR receptor (sTfR), ferritin (FRT) and C-reactive protein (CRP) were measured in venous blood samples from 131 random patients admitted to the ICU for at least 24 h (Length of ICU stay, LIS; min: 1 day; max: 38 days). RESULTS: Anaemia (Hb < 12 g/dL) was present in 76% of the patients (Hb < 10 g/dL in 33%), hypoferremia (SI < 45 microg/dl) in 69%; satTRF < 20% in 53%; FRT < 100 ng/mL in 23%; sTfR > 2.3 mg/dL in 13%; and CRP > 0.5 mg/dL in 88%. Statistically significant correlations (r of Pearson; *p < 0.05, **p < 0.01) were obtained for serum CRP levels and WBC**, Hb*, TRF**, satTRF*, and FRT**. There was also a strong correlation between TRF and FRT (-0.650**), but not between FRT and satTRF or SI. LIS correlated with Hb*, CRP**, TRF*, satTRF* and FRT**. CONCLUSIONS: A large proportion of critically ill patients admitted to the ICU presented the typical functional iron deficiency (FID) of acute inflammation-related anaemia (AIRA). This FID correlates with the inflammatory status and the length of stay at the ICU. However, 21% of the ICU patients with AIRA had an associated real iron deficiency (satTRF < 20; FRT < 100 and sTfR > 2.3). Since oral supplementation of iron seems to be ineffective, all these patients might benefit of iv iron therapy for correction of real or functional iron deficiency, which in turn might help to ameliorate their inflammatory status. PMID- 15813396 TI - [Malnutrition prevalence in patients admitted to a rehabilitation and orthopedic surgery hospital]. AB - OBJECTIVE: Different studies show the scarce attention granted to the nutritional state in historical and clinical practice, what determines the ignorance of the patient's nutritional status to their entrance in the hospital and, therefore, the impossibility to prevent the hospital malnutrition. The objective of our study has been to determine the prevalence of patients' malnutrition entered in a Hospital of Orthopedic surgery and Rehabilitation. METHODS: Observational and analytical study in 250 randomized patients (60% men and 40% women), who were nutritionally evaluated when entering, the hospital, by means of anthropometry (Weigh, height, BMI, skin-fold, corporal circumferences) and biochemical tests (Albumin, Prealbumin and Transferrin). RESULTS: According to the BMI the prevalence of malnutrition was of 8%, the average of caloric malnutrition was of 2.8% (according to anthropometry) and the prevalence of many-sided malnutrition rises to 54.8% (according to biochemical markers). CONCLUSIONS: The high prevalence of fundamental malnutrition (54.8%) demonstrated in this study, it shows the importance of determining the nutritional status when entering the hospital, particularly surgical type' patiens as is the case of most of those who enter the Orthopedic surgery hospitals. PMID- 15813397 TI - [Enteral prostheses for palliative treatment of gastroduodenal obstruction: endoscopic procedure]. AB - Malignant gastroduodenal obstruction may cause significant morbidity. Insertion of enteral prostheses as a palliative treatment is an acceptable option to achieve an appropriate gastric voiding without the morbidity associated to palliative surgery, with a lesser procedure-related mortality and a lower cost, while providing a better quality of life. Its is important to unify and consolidate the endoscopic procedure for the placement of enteral prostheses, avoiding variations that may preclude an adequate analysis of this procedure outcome. PMID- 15813398 TI - [The importance of functional ingredients in pediatric milk formulas and cereals]. AB - Feeding during the first months of life has a decisive influence not only in early developmental phases of children but also in their long-term health. Therefore, inclusion of new functional ingredients to pediatric milk formulas is, nowadays, the matter of a number of studies, incorporating day-by-day newer products and more similar to human milk, the gold standard, which contains them in a natural way. Nowadays, technological advances have made possible to obtain these ingredients from animal or vegetal sources for their addition into pediatric milk formulas and cereals. In the present article, the main functional ingredients, such as polyunsaturated fatty acids, palmitate, nucleotides, pre- and probiotic oligosaccharides, carnitine and taurine are reviewed with regards to their nutritional and functional interest in infant feeding. Their incorporation in pediatric foods has been possible after carrying out animal and clinical studies that provide a solid scientific basis which guaranties their proper bioavailability and their efficacy in the newborn. In the future years, thanks to a better understanding of maternal milk composition and to the performance of further studies, it will be possible to add new ingredients that will allow to obtain pediatric milk formulas more and more similar to maternal milk from a nutritional and functional point of view. PMID- 15813399 TI - [Symbiotics: a new strategy in critically ill patients treatment]. AB - Maintenance of the gut environment is a key factor in determining outcome in the care of critically ill and postoperative patients. It is especially important to maintain both gastrointestinal secretions, full o anti-infectious and anti inflammatory compounds, and the gut flora. Prebiotics, usually polysaccharides, exhibit strong bio-activity and the ingestion of prebiotics has been shown to reduce the rate of infection and restore health in sick and postoperative patients. Probiotics may have at least five functions, all of great importance to the sick patients: the reduction or elimination of potentially pathogenic micro organism of various kinds; the reduction or elimination of various toxins, mutagens, carcinogens, etc.; modulation of the innate and adaptive immune defence mechanisms; the promotion of apoptosis; and the release of numerous nutrient, antioxidant, growth, coagulation and other factors necessary for recovery. A combination of pre and probiotics is referred to as "synbiotics". Our experience of synbotic treatment in critically ill patients is limited, but cutting-edge results from studies of severe acute pancreatitis, chronic hepatitis and liver transplantation offer great hope for the future. This is especially importante as pharmaceutical treatment, including the use of antibiotics, has largely failed, and the medical world is in much need of new treatment paradigms. PMID- 15813401 TI - A parametric feature extraction and classification strategy for brain-computer interfacing. AB - Parametric modeling strategies are explored in conjunction with linear discriminant analysis for use in an electroencephalogram (EEG)-based brain computer interface (BCI). A left/right self-paced typing exercise is analyzed by extending the usual autoregressive (AR) model for EEG feature extraction with an AR with exogenous input (ARX) model for combined filtering and feature extraction. The ensemble averaged Bereitschafts potential (an event related potential preceding the onset of movement) forms the exogenous signal input to the ARX model. Based on trials with six subjects, the ARX case of modeling both the signal and noise was found to be considerably more effective than modeling the noise alone (common in BCI systems) with the AR method yielding a classification accuracy of 52.8+/-4.8% and the ARX method an accuracy of 79.1+/ 3.9 % across subjects. The results suggest a role for ARX-based feature extraction in BCIs based on evoked and event-related potentials. PMID- 15813400 TI - Perceptual limits for a robotic rehabilitation environment using visual feedback distortion. AB - Imperceptible visual distortion, in the form of a disguised progression of performance goals, may be a helpful addition to rehabilitation after stroke and other brain injuries. This paper describes work that has been done to lay the groundwork for testing this hypothesis. We have constructed and validated an experimental environment that provides controllable visual distortion and allows precise force and position measurements. To estimate the amount of visual distortion that should be imperceptible, we measured the limits for force and distance/position perception in our rehabilitation environment for young and elderly unimpaired subjects and for a single traumatic brain injury (TBI) patient. We found the Just Noticeable Difference (JND) for produced force to be 19.7% (0.296 N) and the JND for movement distance/finger position to be 13.0% (3.99 mm) for young subjects (ages 18-35). For elderly subjects (ages 61-80), the JND for force was measured to be 31.0% (0.619 N) and the JND for distance/po'sition was 16.1% (5.01 mm). JNDs of 46.0% (0.920 N) and 45.0% (14.8 mm) were found for the motor-impaired individual. In addition, a subject's rating of effort was found to be profoundly influenced by visual feedback concerning the force magnitude. Even when this feedback was distorted, it accounted for 99% of the variance of the effort rating. These results indicate that substantial visual distortions should be imperceptible to the subject, and that visual feedback can be used to influence the subject's perceived experience in our robotic environment. This means that we should be able to use imperceptible visual distortion to alter a patient's perception of therapeutic exercise in a robotic environment. PMID- 15813402 TI - A simple device to monitor flexion and lateral bending of the lumbar spine. AB - Monitoring compliance with exercise and motivating patients with lower back pain to perform prescribed exercise regimens are considerable tasks. The objective of this study was to develop and test a low-cost device that can be used by a patient at home to both record and provide real-time biofeedback of lumbar position in the midsagittal and frontal planes during exercises. Our device utilizes strain gages on a thin stainless steel beam to measure lumbar flexion extension and an optical mouse sensor attached to the end of the blade to measure lateral bending. In comparison tests with a standard electrogoniometer, our device was shown to be accurate within 3 degrees in both the sagittal and frontal planes in healthy subjects. Furthermore, users were capable of reapplying the device themselves and obtaining measurements that were repeatable within 4 degrees in both planes. The capability of this simple device to accurately measure lumbar spine position in a nonlaboratory setting makes it well suited as a tool for providing feedback on exercise performance to both patients and clinicians. PMID- 15813403 TI - Utilization of ultrasound sensors for anti-collision systems of powered wheelchairs. AB - Anti-collision systems have been developed for use with powered wheelchairs in order to enable people with cognitive or physical impairments to safely operate a powered wheelchair. Anti-collision systems consist of sensors that have the ability to detect objects near the wheelchair and a computer that can stop the chair if a collision is determined to be likely. This investigation considered the suitability of using ultrasound sensors in such a system when encountering objects typically found within a home or a long-term care facility. An ultrasound sensor's ability to detect an object was dependent on the object's size, shape, specularity, reflectivity, and sound absorption characteristics. Ultrasound sensors, by themselves, were found to be unsuitable for anti-collision systems due to an inability to detect objects commonly encountered in the target environment (the home or long-term care facility) without increasing the complexity of the system to such a degree that it would be prohibitive to deploy this technology to the public. PMID- 15813404 TI - Robot-enhanced motor learning: accelerating internal model formation during locomotion by transient dynamic amplification. AB - When adapting to novel dynamic environments the nervous system learns to anticipate the imposed forces by forming an internal model of the environmental dynamics in a process driven by movement error reduction. Here, we tested the hypothesis that motor learning could be accelerated by transiently amplifying the environmental dynamics. A novel dynamic environment was created during treadmill stepping by applying a perpendicular viscous force field to the leg through a robotic device. The environmental dynamics were amplified by an amount determined by a computational learning model fit on a per-subject basis. On average, subjects significantly reduced the time required to predict the applied force field by approximately 26% when the field was transiently amplified. However, this reduction was not as great as that predicted by the model, likely due to nonstationarities in the learning parameters. We conclude that motor learning of a novel dynamic environment can be accelerated by exploiting the error-based learning mechanism of internal model formation, but that nonlinearities in adaptive response may limit the feasible acceleration. These results support an approach to movement training devices that amplify rather than reduce movement errors, and provide a computational framework for both implementing the approach and understanding its limitations. PMID- 15813405 TI - Nonlinear modeling of FES-supported standing-up in paraplegia for selection of feedback sensors. AB - This paper presents analysis of the standing-up manoeuvre in paraplegia considering the body supportive forces as a potential feedback source in functional electrical stimulation (FES)-assisted standing-up. The analysis investigates the significance of arm, feet, and seat reaction signals to the human body center-of-mass (COM) trajectory reconstruction. The standing-up behavior of eight paraplegic subjects was analyzed, measuring the motion kinematics and reaction forces to provide the data for modeling. Two nonlinear empirical modeling methods are implemented--Gaussian process (GP) priors and multilayer perceptron artificial neural networks (ANN)--and their performance in vertical and horizontal COM component reconstruction is compared. As the input, ten sensory configurations that incorporated different number of sensors were evaluated trading off the modeling performance for variables chosen and ease-of use in everyday application. For the purpose of evaluation, the root-mean-square difference was calculated between the model output and the kinematics-based COM trajectory. Results show that the force feedback in COM assessment in FES assisted standing-up is comparable alternative to the kinematics measurement systems. It was demonstrated that the GP provided better modeling performance, at higher computational cost. Moreover, on the basis of averaged results, the use of a sensory system incorporating a six-dimensional handle force sensor and an instrumented foot insole is recommended. The configuration is practical for realization and with the GP model achieves an average accuracy of COM estimation 16+/-1.8 mm in horizontal and 39+/-3.7 mm in vertical direction. Some other configurations analyzed in the study exhibit better modeling accuracy, but are less practical for everyday usage. PMID- 15813406 TI - Gait analysis of people walking on tactile ground surface indicators. AB - Tactile ground surface indicators installed on sidewalks help visually impaired people walk safely. The visually impaired distinguish the indicators by stepping into its convexities and following them. However, these indicators sometimes cause the nonvisually impaired to stumble. This study examines the effects of these indicators by comparing the kinematic and kinetic variables of walking on paths with and without indicators. The results suggest that walking on the indicators causes extra movements of the lower extremities such as increased minimum toe heights during the midswing phase, increased peak hip flex moments, and increased peak hip heights. This study also suggests that a functional leg length discrepancy while walking on the indicators is one of the reasons for the extra movements of the lower extremities. Therefore, we designed a new recessed tactile surface to offset the differences of surface depth while walking on the path containing indicators, and found that the newly designed recessed tactile surface was effective in reducing the extra movements while walking on the path in which indicators were installed. These indicators may help both the visually impaired and elderly people with normal vision to walk safely. PMID- 15813407 TI - Reflex regulation of antagonist muscles for control of joint equilibrium position. AB - A systems model of spinal neuro-musculo-skeletal system (alpha - gamma model) is developed to investigate the plausible roles of spinal proprioceptive feedback in movement control. The model is composed of a joint, a pair of antagonist muscles, length and velocity feedback from muscle spindle, as well as spinal stretch reflex, reciprocal inhibition and recurrent inhibition of Renshaw cells. A descending command modulates the background activation of alpha motoneuron pools in combination with these reflex activities. A static gamma command controls the fusimotor contraction of the spindle. Simulation results reveal that the equilibrium joint angle is linearly correlated to the level of static gamma fusimotor activity of the spindle for a wide range of external loading conditions and reflex gains, suggesting that these spinal reflexes may contribute to regulate the equilibrium position of the joint. Sensitivity analysis further shows that reflex gains and other central commands alter the quasi-linear relation in regular fashions. The reciprocal inhibition gain changes the slope of the linear theta(eq) - gamma curve; and the descending alpha excitation, the stretch reflex gain, and the external load all shift the theta(eq) - gamma curve in parallel. These results imply that reflex gains and descending alpha commands may be coordinated to maintain a unique theta(eq) - gamma curve while providing the flexibility to counteract external loads, to execute a movement, or to regulate additional muscle variables. Dynamic simulation suggests that control of a class of movements can be achieved with a triphasic, alpha pulse and a continuous gamma signal. The model study supports the notion of a dual strategy for controlling trajectories via a feedforward alpha command and for regulating the final equilibrium positions via a feedback gamma command. PMID- 15813408 TI - Closed-loop cortical control of direction using support vector machines. AB - Motor neuroprosthetics research has focused on reproducing natural limb motions by correlating firing rates of cortical neurons to continuous movement parameters. We propose an alternative system where specific spatial-temporal spike patterns, emerging in tasks, allow detection of classes of behavior with the aid of sophisticated nonlinear classification algorithms. Specifically, we attempt to examine ensemble activity from motor cortical neurons, not to reproduce the action this neural activity normally precedes, but rather to predict an output supervisory command to potentially control a vehicle. To demonstrate the principle, this design approach was implemented in a discrete directional task taking a small number of motor cortical signals (8-10 single units) fed into a support vector machine (SVM) to produce the commands Left and Right. In this study, rats were placed in a conditioning chamber performing a binary paddle pressing task mimicking the control of a wheelchair turning left or right. Four animal subjects (male Sprague-Dawley rats) were able to use such a brain-machine interface (BMI) with an average accuracy of 78% on their first day of exposure. Additionally, one animal continued to use the interface for three consecutive days with an average accuracy over 90%. PMID- 15813409 TI - Automated estimation of initial and terminal contact timing using accelerometers; development and validation in transtibial amputees and controls. AB - The aim of this study was to develop and validate an automated accelerometry based system for estimating initial contact (IC) and terminal contact (TC) timing information from walking patterns of healthy control subjects and transtibial amputees that can be used in daily life with minimal interference of researchers. Subjects were instrumented with two uniaxial accelerometers just below the knee while synchronized ground reaction force (GRF) recordings were used as reference measurements. An automated multiphase algorithm was developed to estimate the time of IC and TC in the acceleration signals of five healthy subjects and two transtibial amputees walking at different walking speeds. The accuracy of the detection algorithm in ten control subjects and eight transtibial amputees indicated mean errors ranging between 0.013 and 0.034 s for the TC and IC timing, with 95 % confidence interval of the individual step errors ranging between 0.062 and 0.115 s. Correlation coefficients between the estimated stance phase duration and GRF data were 0.98 and 0.97 for controls and amputees, respectively. We concluded that IC and TC can be accurately and easily measured using this system in both healthy subjects and transtibial amputees walking at different walking speeds. The system can be used in clinical situations or gait labs as well as during daily life. PMID- 15813410 TI - An improved P300-based brain-computer interface. AB - A brain-computer interface (BCI) is a system for direct communication between brain and computer. The BCI developed in this work is based on a BCI described by Farwell and Donchin in 1988, which allows a subject to communicate one of 36 symbols presented on a 6 x 6 matrix. The system exploits the P300 component of event-related brain potentials (ERP) as a medium for communication. The processing methods distinguish this work from Donchin's work. In this work, independent component analysis (ICA) was used to separate the P300 source from the background noise. A matched filter was used together with averaging and threshold techniques for detecting the existence of P300s. The processing method was evaluated offline on data recorded from six healthy subjects. The method achieved a communication rate of 5.45 symbols/min with an accuracy of 92.1% compared to 4.8 symbols/min with an accuracy of 90% in Donchin's work. The online interface was tested with the same six subjects. The average communication rate achieved was 4.5 symbols/min with an accuracy of 79.5 % as apposed to the 4.8 symbols/min with an accuracy of 56 % in Donchin's work. The presented BCI achieves excellent performance compared to other existing BCIs, and allows a reasonable communication rate, while maintaining a low error rate. PMID- 15813411 TI - Investigating the stationarity of paediatric aspiration signals. AB - An aspiration signal is the time-varying anterior-posterior acceleration measured infero-anterior to the thyroid notch when foreign material enters the airway during inspiration. The hypothesis of weak stationarity is tested on aspiration signals by the reverse arrangements test. Results indicate that aspiration signals cannot be uniformly regarded as weakly stationary. Forty-five percent of the examined signals violated the stationarity hypothesis. For these signals, time-varying variance and spectral density structure are identified as major sources of nonstationarity. Stationarity test results generally corroborate qualitative clinical descriptions of aspiration. However, stationarity analysis indicates that aspiration signals are highly heterogenous, a finding which poses significant challenges to the automatic detection of aspirations by accelerometry. PMID- 15813412 TI - Excitability of chronic hemiparetic muscles: determination of chronaxie values and strength-duration curves and its implication in functional electrical stimulation. AB - Central nervous system disorders affect the anatomy and physiology of the lower motoneuron. This fact has an impact on the stimulation parameters, especially on the duration of the stimulating impulses, for functional electrical stimulation in chronic hemiparetic patients. The aim of this study was thus to test the excitability and to determine chronaxie values and strength-duration curves of weak wrist and finger extensor muscles and spastic finger and wrist flexor muscles in the hemiparetic arm. Twelve patients with chronic hemiplegia (>6 months after the onset of the cerebral lesion) participated in the study. A constant current stimulator was used. As to chronaxie values no significant differences were found between the extensor muscles (mean+/-SD: 0.44+/-0.16 ms) and flexor muscles (mean+/-SD: 0.36+/-0.22 ms). A moderate variability was seen for both extensor muscles (0.2-0.8 ms) and flexor muscles (0.1-0.9 ms). These values are well within the normal range determined for innervated muscles. All strength-duration curves were completely normal for each muscle. We conclude that in chronic hemiparetic muscles, impulses of the same duration can be used as in muscles of healthy subjects. PMID- 15813413 TI - Changes in nitric oxide levels in striated muscles of rats following different types of death. AB - BACKGROUND: Nitric oxide is a signal molecule regulating the organism functions in living bodies. The aim of this study was to investigate the NO levels of striated muscles after different types of death in rats. METHODS: Nitric oxide levels in the muscles of masseter, triceps, and quadriceps obtained from right and left sides of 24 Spraque-Dawley rats following death were investigated. The rats were divided into three groups as cervical dislocation (control) group, electric shock group, and drowning group. After applying a light anesthesia, the rats were killed by cervical dislocation, electric shock and drowning. The samples were taken immediately and 120 minutes after death. RESULTS: In all muscle types of all groups, NO concentrations were lower in samples obtained 120 minutes after death than in those obtained immediately after death. NO concentrations were lower in the electric shock and drowning group than in the control group for both times. CONCLUSION: We can conclude that the type of death may affect the occurrence of rigor mortis and NO measurement may give an important clue in evaluation the mode of death. PMID- 15813414 TI - [Gamma-hydroxybutyrate (GHB) misuse or abuse and interpretation of toxicological findings]. AB - Gamma-hydroxybutyrate (GHB) Misuse or Abuse and Interpretation of Toxicological Findings The paper presented is a review of important information on gamma hydroxybutyrate abuse or misuse, its toxicological findings under various circumstances, endogenous concentrations and potentional neoformation, with stressing the difficulties of interpretation which shall be done very carefully. PMID- 15813417 TI - [The Order of Quebec Nurses publicizes its position on the drug policy]. PMID- 15813415 TI - [New legislation on expert witnesses in the Slovak Republic]. AB - Legal framework for expert witnesses in the continental legal system differs from that of common law in many aspects. The position of an expert witness in continental legal system is more formalized and the processual rights and duties of an expert witness differs from rights and duties of witness in general. As of 1st September 2004 the Act No. 417/2002 Coll. on Expert Witnesses, Translators and Interpreters became effective in the Slovak Republic. The Act repealed expert witnesses legislation that originated from 1967 which is still effective in the Czech Republic. The authors present a short overview of the Act. PMID- 15813418 TI - Side effects of anticytokine strategies. AB - Anticytokine strategies probably represent the most important breakthrough in the treatment of inflammatory diseases in the last decade. However, blocking the bioactivity of proinflammatory cytokines, crucial activators of host defence, has proved to be accompanied by an increased susceptibility to infections, especially with Mycobacteria, Salmonellae and fungal pathogens. Multiple mechanisms for these side effects have been proposed, such as inhibition of gamma-interferon production, decreased expression of pattern-recognition receptors, and leucocyte apoptosis. Caution is therefore warranted when these treatments are given to patients with an increased risk for infections. A range of side effects other than infection have been reported. PMID- 15813419 TI - Local and systemic thrombolytic therapy for acute deep venous thrombosis. AB - This article presents a review of the treatment of lower-extremity deep venous thrombosis (DVT) with systemic and catheter-directed thrombolysis (CDT) and percutaneous mechanical thrombectomy (PMT). Standard treatment including anticoagulation therapy and compression stockings may not be entirely adequate, because a significant proportion of patients eventually develop post-thrombotic syndrome (PTS). Thrombolytic agents might offer a potential advantage because they cause faster and more complete clot resolution, which may reduce or prevent residual vein stenosis and valve damage. Thrombolytic therapy results in greater lysis, but also in higher complication rates than does anticoagulation alone. Major bleeding occurs in 11% of patients treated with thrombolytic therapy. The incidence of PTS tends to be lower in patients treated with thrombolytics. However, several methodological flaws limit the conclusions with respect to reduction in PTS. No adequate randomised controlled trials have been performed comparing CDT or PMT with conventional therapy. Given the current data, thrombolytic treatment, CDT or PMT should not be applied except in extraordinary cases. First, the long-term effectiveness in terms of reducing PTS, although possible, remains uncertain. Second, the risks of thrombolytic therapy and PMT are higher. Third, current conventional therapy is relatively inexpensive, convenient and safe. PMID- 15813420 TI - Falls and medications in the elderly. PMID- 15813421 TI - Evaluation of cardiac ischaemia in cardiac asymptomatic newly diagnosed untreated patients with primary hypothyroidism. AB - BACKGROUND: Hypothyroidism is regarded as a risk factor for coronary artery disease. Possible factors involved in this association are hyperlipidaemia and hypertension, both occurring with increased frequency in hypothyroid patients. The aim of our study was to evaluate signs/symptoms of cardiac ischaemia in untreated hypothyroid patients without angina pectoris, since this has never been performed before. METHODS: 51 consecutive cardiac asymptomatic patients (mean age 47, range 22 to 86 years) were studied by dobutamine stress echocardiography and bicycle ergometry. RESULTS: Mean values of body mass index, resting heart rate and blood pressure were 28.5 kg/m2, 68 beats/min and 129/81 mmHg, respectively. Median TSH was 51.9 mU/l, mean FT4 7.3 +/- 2.9 pmol/l (mean +/- SD), TT3 1.6 +/- 0.6 nmol/l and total cholesterol was 5.8 +/- 1.6 mmol/l. None of the patients had symptoms of angina pectoris during dobutamine stress echocardiography or bicycle ergometry and no evidence of myocardial ischaemia was demonstrated. Exercise tolerance, assessed by dividing the maximum achieved workload by the target performance (depending on body height, sex and age), was diminished in 38% of patients, and significantly related to the degree of hypothyroidism. CONCLUSION: No angina pectoris or cardiac ischaemia at exercise or stress was found in cardiac asymptomatic hypothyroid patients. The precise role of hypothyroidism as a risk factor for coronary artery disease should be further elucidated. PMID- 15813422 TI - Shared care with task delegation to nurses for type 2 diabetes: prospective observational study. AB - BACKGROUND: To study the effects of two different structured shared care interventions, tailored to local needs and resources, in an unselected patient population with type 2 diabetes mellitus. METHODS: A three-year prospective observational study of two interventions and standard care. The interventions involved extensive (A) or limited (B) task delegation from general practitioners to hospital-liaised nurses specialised in diabetes and included a diabetes register, structured recall, facilitated generalist-specialist communication, audit and feedback, patient-specific reminders, and emphasised patient education. The target population consisted of 2660 patients with type 2 diabetes treated in the primary care setting. Patients who were terminally ill or who had been diagnosed with dementia were excluded from the study. RESULTS: The participation rates were high (90%) for patients, and none of the 64 GPs discontinued their participation in the study. Longitudinal analyses showed significant improvements in quality indicators for both intervention groups (process parameters and achieved target values on the individual patient level); in standard care, performance remained stable or deteriorated. Both patients and caregivers appeared satisfied with the project. CONCLUSION: This study shows that structured shared care with task delegation to nurses, targeted at a large unselected general practice population, is feasible and can positively affect the quality of care for patients with type 2 diabetes. PMID- 15813423 TI - A patient with dyspnoea, subfebrile temperature and electrocardiographic abnormalities. PMID- 15813424 TI - Tonsillar tuberculosis in a rheumatoid arthritis patient receiving anti-TNFalpha (adalimumab) treatment. AB - This case report describes a 61-year-old rheumatoid arthritis patient with an atypical clinical presentation of a sore throat. Because of rheumatoid arthritis refractory to conventional disease-modifying antirheumatic drug therapy, anti TNFalpha was felt to be indicated, and a screening for tuberculosis was carried out. As the screening for tuberculosis (PPD) was positive, isoniazid was prescribed prophylactically for six months. After eight months of anti-TNFalpha (adalimumab) treatment, he developed tonsillar enlargement and nodular pulmonary lesions. Histopathological and microbial investigations established the diagnosis of tonsillar tuberculosis. PMID- 15813425 TI - Isolated perianal tuberculosis. AB - Perianal tuberculosis, without the presence of any previous or active pulmonary infection, is extremely rare. A case of isolated perianal tuberculosis without gastrointestinal or pulmonary spread will be discussed here with an evaluation of the clinical features. PMID- 15813426 TI - [Icterus prolongatus is a reason for laboratory investigation, also in breastfed neonates]. AB - Three infants, a boy aged 4 months and two girls aged 3 months and 6 weeks, respectively, had jaundice while they were breastfed. Until then, the jaundice had been interpreted as an innocent consequence of the breastfeeding. In the two eldest patients, however, biliary atresia was diagnosed. A hepatoportoenterostomy was performed in the girl when she was 15 weeks old, but both ultimately underwent a liver transplantation with a good clinical outcome. In the youngest patient, the jaundice disappeared spontaneously and retrospectively was indeed probably associated with breastfeeding. Thorough physical examination and biochemical analyses (total and direct bilirubin, gamma-glutamyl-transferase) are important for the identification of neonatal cholestasis syndromes. Laboratory investigation is recommended in any neonate jaundiced after the age of 3 weeks to differentiate pathological neonatal cholestasis from prolonged jaundice related to breastfeeding. PMID- 15813427 TI - [The value of gene expression profiles using micoarrays for the individualisation of adjuvant therapy after surgery for breast cancer]. AB - Gene-expression profiles are a promising development in determining the prognosis of patients with primary breast cancer. They accurately assess the risk on distant recurrence of disease and show if the patient might benefit from adjuvant therapy after surgery thus minimising the risk of distant metastases. Three clinically relevant profiles on prognosis have now been published, two of which come from the Netherlands, and whose results are an improvement on those using traditional clinical parameters i.e. the St. Gallen and the National Institutes of Health criteria. These gene-expression profiles mean that 25-40% of patients need no longer undergo adjuvant systemic therapy (chemotherapy and/or endocrine therapy). Although the risk-stratifying power of these profiles has been established, their power in predicting the response of the patients to adjuvant systemic therapy still awaits scientific proof. PMID- 15813428 TI - [The diagnosis of acute myeloid leukaemia enhanced by using DNA microarrays]. AB - Recently, two studies have shown that the use ofgene-expression profiling using DNA microarrays or DNA chips may improve the classification of acute myeloid leukaemia (AML). In both studies, cluster analyses based on the molecular signatures defined known subgroups as well as novel subgroups of AML. Chromosomal lesions, mutations, and abnormal gene expression with prognostic value determined the clustering. In fact, gene-expression profiling recognized leukaemias with certain chromosomal aberrations that had been missed by routine cytogenetics. Thus, gene-expression profiling allows a comprehensive classification of AML that includes previously-identified genetically-defined as well as novel prognostically-relevant subgroups. One comprehensive DNA chip may in the future replace a variety of cytogenetic, immunological and molecular techniques that are currently used in combination. PMID- 15813429 TI - [Molecular unraveling of disease by means of DNA-microarrays]. AB - Determination of the human genome sequence and the development of microarray technologies allowing the rapid measurement of all genes in the genome have generated new perspectives for our current biomedical research. Gene expression analysis will make a major contribution to our insight into the underlying biology of disease and will lead to improved methods for diagnostics, prognosis and treatment. Microarray studies create the possiblity to subclassify patients with diseases such as rheumatoid arthritis, diffuse large B-cell lymphoma and breast cancer, with both prognostic and therapeutic consequences. The simultaneous quantification of the activity of all genes in tissues or cells from patients by microarray technology, linked to the clinical parameters, creates a large number of data points, which cannot be analysed without the aid of the advanced application of bioinformatics. As a result, genomic research has become, in part, a bioinformatics discipline that will be integrated with clinical medicine. The microarray technology makes it possible to develop personalized medicine, with a more accurate diagnosis and prognosis for every patient and subsequently a tailored treatment strategy. PMID- 15813430 TI - [Measuring the length of newborn infants]. AB - Since 1985, measurement of the length of neonates has been practically abandoned in the Netherlands because it was thought that stretching the legs and knees briefly in order to measure the length immediately after birth could be harmful for the development of the hip joint. However, this fear seems unjustified. Measuring the length of the neonate provides useful information regarding the general condition and has predictive value for the final adult height. If an infant is disproportionately small in comparison with its weight or small for its gestational age with insufficient catch-up growth, this may be an indication of underlying pathology. As a rule, the length of all neonates should be measured immediately after birth and length measurement should be re-introduced as part of standard care. Length measurement can be done with sufficient accuracy after proper instruction. For babies born after incomplete breech presentation, length measurement should be postponed for about a week. PMID- 15813431 TI - [Diagnostic image (232). A neonate with bilious vomiting]. AB - A prematurely born girl presented with abdominal distension, bilious vomiting and a football sign on a plain abdominal X-ray, due to accumulation of free gas following infarction and perforation of the ileum: a complication of indomethacine. PMID- 15813432 TI - [Pneumatosis intestinalis in 9 children with an oncologic disease]. AB - OBJECTIVE: To acquire knowledge regarding the rare condition pneumatosis intestinalis (PI) in children treated for malignant disease. DESIGN: Retrospective. METHOD: In 1998-1999 PI was diagnosed in 9 of the 140 children with malignant disease in the department of Paediatric Oncology of the UMC St Radboud, Nijmegen, the Netherlands. By examination of the records of these 9 children, data were collected on the symptomatology, diagnostics, treatment and prognosis of PI. RESULTS: The 9 children included 7 boys and 2 girls, varying in age from 2 to 12 years. In 7 patients the underlying disease was acute lymphocytic leukaemia and in 2 it was a stage IV neuroblastoma. The presenting symptoms were nonspecific and included: a distended abdomen, abdominal pain, diarrhoea and constipation. In all children, PI was located in the colon. Supplemental blood and microbiological analysis did not reveal any typical abnormalities. 8 children were treated with lactitol because of constipation. A laparotomy was performed in the first patient, while the other 8 were treated with gastric suctioning, parenteral nutrition and antibiotics. All 9 children recovered within a few weeks. CONCLUSION: With supportive care, PI in children with malignant disease is mostly a self-limiting condition. A pneumoperitoneum in PI is no indication for surgery, except in the presence of an acute abdomen. Chemotherapy can be continued. PMID- 15813433 TI - [Eosinophilic oesophagitis in young men with food swallowing complaints]. AB - Three men, aged 20, 24 and 42 years, reported difficulties in passing food through the oesophagus. The diagnosis of eosinophilic oesophagitis was made after endoscopic investigation, laboratory tests and histological tests. In all three patients the symptoms disappeared: respectively spontaneously, during systemic treatment with corticosteroids due to a kidney complaint, and after topical corticosteroid treatment lasting 6 weeks. Eosinophilic oesophagitis occurs in particular in young men. There are complaints about the passage of food through the oesophagus, with frequent food impaction, also without any obvious stenosis. Endoscopic features are subtle and comprise a vulnerable oesophageal mucosa with a ringed appearance or small white spots on the oesophageal mucosa. Histopathology reveals an eosinophilic infection infiltrate in the oesophageal epithelium. Food allergies may play a causal role. With respect to the treatment, favourable results have been described for oral fluticasone, while endoscopic treatment may consist of dilation. PMID- 15813434 TI - [Hoarseness in a recent visitor to the tropics through infection of the larynx by Histoplasma capsulatum]. AB - A 58-year-old man presented with hoarseness and a sore throat for the previous 3 months, which were caused by a laryngeal infection with the fungus Histoplasma capsulatum. He had been infected during a stay in Indonesia. Both his hoarseness as well as the Histoplasma antigen titres in serum responded to therapy with itraconazol. H. capsulatum is found in tropical and subtropical areas worldwide, but infections are most commonly reported from the south of the United States of America. If not asymptomatic, the disease most often presents with pulmonary symptoms. A histoplasmosis presenting as a laryngeal infection is relatively uncommon and can be mistaken for papillomatosis or carcinoma. The diagnosis can be made by microscopy and culture of the fungus; an antigen test is available in the United States. PMID- 15813435 TI - [The 'acute scrotum' in children: the clinical presentation as indicated by a rapid operation]. PMID- 15813436 TI - [When the treatment in children with asthma fails: first other reasons should be sought before prescribing more medication]. PMID- 15813437 TI - [When the treatment in children with asthma fails: first other reasons should be sought before prescribing more medication]. PMID- 15813438 TI - [Intimate partner abuse of women: identification of victims in medical practice]. PMID- 15813439 TI - [Deliberate termination of life in newborns in The Netherlands; review of all 22 reported cases between 1997 and 2004]. PMID- 15813440 TI - TRACS, an electronic record-keeping system for sedation. PMID- 15813441 TI - Sedation records: TRACS electronic database. AB - The volume of data that should be recorded before, during and after a conscious sedation procedure makes accurate and complete manual recording impractical. Unless the data is in a structured electronic format, subsequent analysis is extremely difficult. The TRacking and Analysis of Conscious Sedation electronic database (TRACS) is designed to overcome these problems. The user is guided through the recording procedure, which follows the Guidelines for Conscious Sedation in the Provision of Dental Care. This, together with the 'Audit Trail' of events provided by TRACS could prove invaluable in the event of a legal or technical dispute. Since the information is in an electronic database, immediate analysis can be performed on both individual and groups of patient records. Provision has been made for records from many treatment centres to be consolidated thus enabling serious academic work to be performed on large numbers of records. PMID- 15813442 TI - Regulation of acupuncture. Update June 2004. PMID- 15813443 TI - The Eastman Dental Institute course in conscious sedation. PMID- 15813444 TI - Sedation in children. PMID- 15813445 TI - Conscious sedation in dental practice in the USA. An update on drugs, techniques and monitoring. PMID- 15813446 TI - Clinical audit. How to identify priority topics. PMID- 15813447 TI - ADA summer scientific meeting, July 2004, Oxford. PMID- 15813448 TI - Crystal balls. An opinion article by Dr. Arthur Jack. PMID- 15813449 TI - [Tension gastrothorax as a late consequence of an undiagnosed rupture of the diaphragm]. AB - The authors present a case-review of a 20-year-old male who suffered from a car accident polytruma (the ISS 22, brain contusion, lung contusion, serial ribs fructures, fracture of the Th8, haematoma of the left kidney with haematuria). Surgery was not indicated, his thorax and abdomen were repeatedly examined with x rays, ultrasound and CT scans, during the patient's hospitalization and during the follow-up period. After some time, the stomach expansion in the pleural cavity imitated the clinical and roentgenological findings typical for the tension pneumothorax. Urgent drainage solved the cardiopulmonary failure and cleared up the diagnosis. This atypical case is followed by a short literature overview. PMID- 15813450 TI - [Could the peroperative manometry of the oesophagus be used for prediction of dysphagia following antireflux procedures?]. AB - A possible development of the postoperative persisting dysphagia, which decreases the quality of life of the operated, remains the main drawback of the laparoscopic antireflux surgery. Among several variations of the antireflux surgical procedures, there is none known to completely eliminate this risk. In this study, supported by the IGA MZ CR ND 7142-3 grant, peroperative measurements of the lower oesophageal sphincter (LES) tone with a concurrent mechanical callibration of the cuff using an oesophageal tube were taken. A complete Nissen Rossetti cuff was applied in 39 patients. Peroperative increases in the LES tone following the completion of the antireflux cuffs were monitored. The patients continued to be monitored with the aim to detect the onset of dysphagia. In the patient group with the peroperative LES tone increase exceeding 15 mmHg, significantly higher rates of prolonged dysphagia were recorded. In cases of lower LES increases, the rates of dysphagia were low and good functioning to prevent any pathological gastrooesophageal reflux was maintained. No complication with respect to the peroperative oesophageal manometry was recorded. Duration of the antireflux operation conducted with the peroperative manometry was prolonged by 4 minutes, on average. Based on the assessment of the data, the authors demonstrate that the LES tone increase on its own, does not provide for the antireflux efect of the fundoplication procedures and that the patients cannot benefit from its extremely high values. On the contrary, its high values may indicate possible risks of dysphagia in the postoperative period. PMID- 15813451 TI - [Injuries to the biliary tract during cholecystectomy]. AB - Injuries to the biliary tract during both the laparoscopic or the open cholecystectomic procedures, remain among the most serious iatrogenic injuries with high morbidity and mortality rates. The higher the number of the laparoscopic cholecystectomies, the higher the number of the injuries to the biliary tract. Early peroperative recognition of these injuries is a prerequisite for successful biliary tract reconstructions. Mucosal hepaticojejunoanastomosis according to Roux is the golden standard of the reconstrucion treatment. Stenoses in anastomoses followed by development of cholangitides are considered serious postoperative complications. In these cases, endoscopic and transparietal dilation with plastic stents implantation is the method of choice. Technically exacting reoperations then follow. The authors present a trial group of 11 patients, who were treated in the Surgical Clinic of the Faculty Hospital in Plzen for biliary tract injuries during cholecyctomic procedures (8 were laparoscopic and 3 open), from 01-01-2000 to 01-09-2004. Severe inflammatory changes in the region of the Calot triangle, were the commonest cause of the biliary tract injuries during primary operations. In most cases (N = 8) the injury was diagnosed and managed immediately during the primary procedure. Hepaticojejunoanastomosis according to Roux was the principal procedure used to repair the biliary tract (N = 8). Postoperative morbidity reached 36.4%, 2 elderly patients exited (18.2%) due to septic multiorgan failure on the 15th day and the 7th month after the surgical procedure. Multidisciplinary approach of a team of experienced surgeons, endoscopists and radiologists in the hepatobiliary region is a fundamental prerequisite for long-term successful outcomes of technically exacting reconstructive procedures of the hepatobiliry tract. PMID- 15813452 TI - [Surgical treatment of distally located rectal carcinomas using the rendez-vous method--a laparoscopic resection in combination with T.E.M]. AB - The aim of this work is to introduce a surgical procedure, which would make operations of distally located rectal carcinomas indicated for abdominoperineal extirpation of the rectum possible while maintaining intestinal continence and full functioning of the sphincter system. At the same time, we have aimed at the maximum use of all pros of contemporary miniinvasive surgical techniques. We have labelled our proposed surgical technique the "rendez-vous technique". The trial group includes the original group of 10 patients, who have been operated in our clinic since April 2004, using the rendez-vous technique. We are aware of the fact, that the trial group is small, however we believe that this trial group is the starting group, which will continue to enlarge and that it will bring results to the patients themselves, as well as for a valid prospective study in order to confirm or challenge the proposed method's effectiveness. PMID- 15813453 TI - [Anatomical resection for liver metastases of the carcinoma of the large intestine and the rectum]. AB - Resection of the colorectal carcinoma liver metastases is an effective therapeutical procedure with a five-year survival rate in 20-50% of the operated. Opinions on the most optimal type of the resection procedure remain to be controversial. The aim of this study was to assess all indication criteria for anatomical resections of the colorectal carcinoma liver metastases. SUBJECTS AND METHODOLOGY: This retrospective study analyzed 98 patients with liver resection for colorectal metastases. In 31 patients, extensive anatomical resections of the liver were conducted, segmentotomie were conducted in 20 patients and nonanatomical wedge resections in 47 patients. Extensive anatomical resections were conducted in cases of tumorous foci larger than 2cm and located marginally between individual segments of a single lobe, and in cases of multiple tumorous foci affecting a single liver lobe. The nonanatomical resections were conducted in cases of metastases smaller than 2 cm, localized on the liver surface or in cases of multiple peripheral foci, including cases when both liver lobes were affected. RESULTS: In the group with the wedge-type resections, a histologically positive border was found in 4 cases. Postoperative complications were recorded in 10 (19.6%) patients with the anatomical resection and in 9 (19.1%) with the wedge resection. CONCLUSION: 1. Anatomical resection of the liver for the colorectal carcinoma metastases is indicated in cases of larger foci (over 2 cm), located marginally between segments or in multiple metastases affecting a single liver lobe. 2. Anatomical approach to the resection lowers the rates of histologicaly non-radical resections (R 1). 3. The new surgical technique of the liver transsection lowered, even in cases of anatomical resections, the postoperative morbidity rates. PMID- 15813454 TI - [The cystadenocarcinoma of the appendix]. AB - The authors present a case-review of a patient with a large retroperitoneal tumor. A right-sided hemicolectomy with a removal of a large retroperitoneal cyst extending behind the liver up to the diaphragm on the right side, was conducted. Histology confirmed a mucous cystadenocarcinoma of the appendix. Furthermore, the authors discuss this fairly rare type of the GIT carcinoma. This tumor is low invasive, mucus producing and rarely metastazing. During the tumor progression, the tumorous cells enter the peritoneal cavity and cause, so called, pseudomyxoma peritonei. The treatment principle is to complete a radical surgical removal of the tumor and, furthermore, in case of the pseudomyxoma periotenei, also application of the local peroperative chemotherapy. PMID- 15813455 TI - [Hemipelvectomy for a desmoidal tumor]. AB - A case- review describing a desmoid in a young female. The tumor originated at the tendinuous attachment of the m. rectus abdominis on the pubic bone, which had had two relapses. During the third--so far the latest--procedure it required left sided hemipelvectomy with resection of the rectum and a part of the vagina. The patient was instructed about the serious character of the disease, the requirement for the radical resection and about the requirement for a careful long-term follow-up of the patient after the procedure. PMID- 15813456 TI - [A case of an asynchronic triple tumorous disorder: a rectal adenocarcinoma, a carcinoma of the kidney and a prostatic adenocarcinoma--case report]. AB - The authors describe occurence asynchronic tumor triplicity. In the year 1980 in 56 years old patient had histologically proven rectal adenocarcinoma and consequently was done radical Miles amputation of rectum. In December 1991 in the same patient was histologically proven well differentiated adenocarcinoma of prostate after transurethral resection of prostate. Prostate cancer was threated bilateral orchiectomy (March 1992) and consequently from April to June 1992 was done small-volume irradiation of pelvic by Betatron X-ray, box technique with dosage 70 Gy. In November 1992 there was done radical transperitoneal nephrectomy on right side for renal carcinoma. Histologically was proven moderately differentiated clear cell renal carcinoma. In August 1994 was found suspected pulmonary metastases, wich was proof on CT in the January 1995. Therapy was Imodin, Wobenzym, Vinblastin. Patient was died of generalisation renal cancer in the February 1998. The carcinoma of prostate and rectum wasn't found in the autopsy. The authors emphasize pertinence radical surgical access incuding multiplex malignant tumors and consider to carry out oncology screening in the all of patients with proven malignant tumor. PMID- 15813459 TI - [Oesophageal perforations]. AB - The subject of oesophageal perforations is wide enough to make any comparison of therapeutical methods and their respective results very difficult, if not impossible. Therefore, the authors aim to specify individual perforation types and their therapeutical management. PMID- 15813457 TI - [Reconstruction surgical procedures for iatrogenic injuries to the ureter. Examples of uncommon, however successful approaches to their management]. AB - The authors present several less usual reconstructions of ureters after the primary iatrogenic lesions. The appropriate ureteric capacity to drain the urine after previous injury is quite demanding and hence before the definitive decision is made the wholesome health status of the patient is to be considered and all possible solutions should be weighted. The efficient primary urinary diversion above the lesion is unavoidable. The genuine reconstruction could be deferred by several weeks or months in the view that in some cases the least invasive (endourologic) methods can completely avoid the need of open surgery in difficult adhesions after previous major surgery. PMID- 15813460 TI - [Laparoscopic diagnosis and treatment of organic hyperinsulinism--our first experience]. AB - Laparoscopic pancreatic procedures are still at the stage of evaluation with regard to their indications and the technical variation used. Laparoscopic pancreatic surgery is currently used for staging malignant pancreatic tumours, for the resection of benign pancreatic tumours and for the occasional management of inflammatory disorders of the pancreas. Insulinomas are rare tumours with incidence per year of 0.1-0.4 per 100,000. Ist Surgical clinic, IIIrd Internal clinic and Radiological clinic are cooperating since 1971. On the list of IIIrd Internal clinic are 80 patients with insulinoma. Since 1971 at Ist Surgical Clinic 66 patients have been operated. In our article are diagnostic and therapeutic problems discussed. Laparoscopic pancreatic procedures are still at an evaluation stage regard to their indications and techniques. Authors give report about first experience with successful laparoscopic enucleation of insulinoma in case 83 year old woman (Fig. 3, 4). The operative time was 90 min. Drainage is necessary. CT guided drain in cavity of seroma is shown (Fig. 5). PMID- 15813461 TI - [Laparoscopy in the colorectal carcinoma treatment]. AB - AIM: The authors present their experience and results with laparoscopic surgery in the colorectal carcinoma treatment in the Centre of Miniinvasive Surgery of the Podlesi-Trinec hospital. METHODOLOGY: In this retrospective study, the authors present a group of 148 patients with colorectal carcinomas operated by their team from 1st January, 2002 to 30th June, 2004. RESULTS: The authors completed resection procedures in 133 patients. In 15 patients, with respect to the extent of their disorder, the authors had to apply derivation colostomies, to construct by-pass ileotranversal anastomoses or to complete the procedures with exploration laparoscopies. CONCLUSION: Thank to miniinvasive techniques, laparoscopy has found its place also among the colorectal carcinoma treatment methods. The presented results, in accordance with other studies published in literature worldwide, present this method as a suitable one for a patient and fulfilling all criteria of the onco-surgical radicality. PMID- 15813462 TI - [Urgent management of the complex pelvic fractures]. AB - AIM OF STUDY: The term "complex pelvic fractures" is reserved for pelvic ring fractures associated with soft tissue injuries in the pelvic region and with haemodynamic instability of the patient. These fractures represent only 10% of all pelvic fractures; however, the morality rate exceeds 33%. This study describes the algorithm of the urgent treatment of serious pelvic ring injuries. METHODS: Urgent application of pelvic clip or external pelvic fixator is considered as a part of the reanimation period. If the external pelvic ring fixation is not followed by stabilization of patient's circulation, the pelvis packing using the lower middle incision without opening of the abdominal cavity must be executed within a short time. The absence of the intraabdominal bleeding is essential. If the pelvis packing does not improve patient's haemodynamic conditions, the bleeding should be controlled by angiography and embolization or by direct ligature of the internal iliac artery. In the most severe cases, hemipelvectomy would be carried out as the life-saving procedure. DISCUSSION: Extreme blood loss associated with complex pelvic ring fractures is possible due to enlarged volume of the injured pelvis. This is caused by a gap within the symphysis or in the region of the pubic rami and by a shift in the posterior pelvic segment. In such situation, waiting for the self-packing of pelvis by haematoma can be dangerous. In case of parapelvic compartments disruption, the effect of the self-packing would not come at all. The management of the pelvic ring fractures should be completed by a surgical procedure, because non-surgical treatment is mostly disappointing. The best results can be achieved only by the early operation of the anterior and posterior pelvic segment aimed at the realignment of pelvic ring and proper anatomical relations. CONCLUSIONS: Complex pelvic ring fractures require the aggressive approach. The basic reanimation procedures include application of the pelvic clip. This is the only way how to control extensive haemorrhage associated with the pelvic ring fractures and how to improve patient's haemodynamic conditions. The final surgical treatment of the pelvis fractures should be completed early after the injury. This is within the competence of specialized centres. PMID- 15813463 TI - [Femoral head replacement in intracapsular femoral neck fractures]. AB - Hemiarthroplasty has still its place in the therapeutic algorithm of the treatment of dislocated intracapsular femoral neck fractures. As compared to internal fixation it is accompanied by less complications and allows an immediate postoperative weight bearing. In contrast to total hip arthroplasty, this operation is not so demanding for the patient. However in the long-term perspective it is often associated with erosion of acetabulum. Therefore it is indicated mainly for biologically older patients with less physical activity without osteoarthritis of the hip. In these patients we usually prefer a cemented monoblock hemiarthroplasty. In case of biologically younger patients where it is impossible to use total hip arthroplasty due to general or local condition, modular hemiarthroplasty is indicated with a removable head allowing in case of acetabular erosion an easy conversion to total hip arthroplasty. Hemiarthroplasty must be correctly indicated and technically properly performed. First of all it is necessary to respect the relation between the centre of the prosthetic head and the apex of the greater trochanter, i.e. the head centre should be 1-2 mm below the level of apex of the greater trochanter. Further, it requires a correct anteversion and suture of the articular capsule as a prevention of postoperative dislocation. PMID- 15813464 TI - [General surgery is not dead. Continuation of the discussion initiated by Dr. P. Pafko]. PMID- 15813466 TI - [Role of the imprimatur]. PMID- 15813467 TI - [Report on a study visit in the USA]. PMID- 15813468 TI - [The founding of the Prague Sanitarium in Prague Podoli]. PMID- 15813470 TI - [In vivo effect of endometria's and human cervix's secretions on the spermatozoid]. AB - OBJECTIVE: To demonstrate the capacitance stages acquired by the spermatozoa after the in vivo contact with the endometrium and the human cervical canal. STUDY TYPE: Experimental, prospective, and longitudinal. MATERIALS AND METHODS: Spermatozoa obtained from the cervix and endometrium, plus post-capacitated semen in a 1 and 4 hours process, from couples that have proved fertility, were processed to evaluate motility, morphology, vitality and identification of morphological patterns with chlortetracycline and then inducted to acrosomal reaction with ionophore A23187. RESULTS: A safe, fast and harmless technique to obtain endometrial secretions is described. The spermatozoa capacitated in vitro and in vivo developed specific capacitance stages, applying Perry's patterns with the CTC test. Spermatozoa capacitated in vitro and then recovered in the cervix responded favorably to the induction of the acrosomal reaction with calcium ionophore A23187, differing from the obtained with endometrium wash. CONCLUSIONS: The spermatozoa recovered from the endometrium presented a more advanced capacitance stage than that obtained from the cervix, this is demonstrated with the induction of the acrosomal reaction and the CTC test. These findings suggest that endometrial and cervical tissues promote capacitance, different in each one of them and progressive in their biological route. PMID- 15813469 TI - [Characterization of choriodecidual space as an effector molecule-rich environment that induces rupture of fetal membranes during labor]. AB - OBJECTIVE: To identify a specific microenvironment in direct contact with fetal membranes where effector molecules acumulate, aiming to degrade the components of its extracellular matrix during labor. TYPE OF STUDY: Experimental, analytic, longitudinal and prospective. MATERIALS AND METHODS: Blood samples were collected from maternal, fetal and choriodecidual compartments, and mononuclear cells were isolated. Part of these cells was stained with antibodies to determine leukocyte subpopulations by flow cytometry. The other part was cocultured for 12 h with amniochorion explants. After coculture, MMP-9 was identified on the mononuclear cells by immunofluorescence. IL-1beta and TNF-alpha were determined in the supernatants by ELISA. Three independent experiments were carried out with duplicates and analyzed with Mann-Whitney's U test. RESULTS: Although there were no significant differences in the mononuclear cell subpopulations from the three compartments, MMP-9 production was higher in choriodecidual cells than in those of the maternal and fetal compartments. Furthermore, IL-1beta and TNF-alpha were significantly more abundant in cocultures with choriodecidual cells compared with the other two compartments. CONCLUSIONS: During labor, choriodecidual cell subpopulations are not phenotypically different from those of the maternal or fetal compartments, but they are regarding MMP-9 production, which suggests that the environment surrounding chorioamniotic membranes enhances the synthesis of this enzyme, thus promoting degradation of connective tissue. PMID- 15813471 TI - [Connective tissue degradation in human amniochorion after stimulation with choriodecidual lymphocytes infected with group B streptococci]. AB - OBJECTIVE: To identify whether soluble products from choriodecidual blood cells stimulated with group B streptococci (GBS) induce connective tissue degradation in human amniochorion. MATERIAL AND METHODS: Blood samples from choriodecidual compartment were collected by direct aspiration from placental cotyledons draining blood and represent local circulating cells. Samples were divided into two aliquots: one was stimulated with GBS (1 x 10(6) CFU/mL) and the other was kept free of bacteria as negative control. After overnight incubation, plasmas were separated. Chorioamnion explants were stimulated with 10% plasma for 12h at 37 degrees C in 5% CO2. MMP-9 proteolytic activity was measured in the supernatants by gelatin-zymography and IL-1beta and TNF-alpha were quantified by ELISA. Distribution of the collagenous fibrils in explants was examined by electron microscopy. STATISTICAL ANALYSIS: three independent experiments on duplicate were carried out and the statistical significance of experimental differences between groups was assessed with ANOVA test. RESULTS: MMP-9, IL-1beta and TNF-alpha production was significantly higher in supernatants from explants co-cultured with choriodecidual plasma from blood previously infected with GBS, compared with control plasma. Accompanying extensive changes of connective tissue arrangement confirm induction of extracellular matrix degradation. CONCLUSIONS: Choriodecidual plasma from blood stimulated with GBS is enriched with biochemical signals that enhance the MMP-9, IL-1beta and TNF-alpha production by amniochorion. These findings suggest that local circulating cells are capable to act in response to GBS choriodecidual infection through extracellular matrix degradation and the consequent rupture of membranes. PMID- 15813472 TI - [Perinatal result of an institutional assisted reproduction program]. AB - BACKGROUND: Assisted reproductive technology manipulates masculine gametes, embryos and implantation. It also aids the known or unknown factor of sterility without having the base problem correction as a target. In vitro fertilization and embryo transfer are among these techniques. OBJECTIVE: To describe the overall outcome and the final perinatal offspring after in vitro fertilization cycle in an institutional third level hospital. MATERIALS AND METHODS: IVF cycles were retrospectively analyzed from October 1999 to May 2004. Several variables were described like: age, fertilization rate, implantation and pregnancy rate, fetal status, time of gestation during labor, miscarriage follicle-stimulating hormone rate and take-home baby rate. Patients underwent hypophyseal supression with GnRH analog, using a long luteal phase protocol and stimulated with recombinant FSH. Overall data is expressed as average +/- standard deviation and percentage. RESULTS: 365 cycles were analyzed in 314 patients, average age was of 34 +/- 3.7 years, tubal factor was diagnosed in 63.3%, fertilization rate was of 60.4%, implantation rate of 37.1%, per transfer pregnancy rate of 25.1%, per transfer live born rate of 21.7%, multiple pregnancy rate of 29.3%, miscarriage rate of 28% and ectopic pregnancy rate of 4.8%. In 87.8% of the cases caesarean operation was made; multiple pregnancy offspring weighted more than 1250 g in 70% of them; 70.5% was born after 32 weeks of pregnancy; 90% was born live and with good prognosis. Glucose metabolism alterations were the most frequent maternal complication. CONCLUSION: Although the results obtained were similar to those of assisted reproduction centers, it suggests improving multiple pregnancy rate and abortion rate. PMID- 15813473 TI - [Effect of pelvic floor exercise during pregnancy and puerperium on prevention of urinary stress incontinence]. AB - BACKGROUND: The main factor for the appearance of urinary stress incontinence in almost all women is pregnancy and vaginal childbirth. The pelvic floor exercises have been described for the treatment of urinary stress incontinence. OBJECTIVE: To determine with a randomized controlled trial if the pelvic floor exercises during pregnancy and late puerperium diminish the prevalence of urinary stress incontinence at 28 and 35 gestational week and at 6 weeks after childbirth. MATERIAL AND METHODS: The study was made with nulliparous, pregnant women who realized pelvic floor exercises during pregnancy and after birth. 72 women were studied, 52.7% realized pelvic floor exercises and 47.2% did not. RESULTS: The urinary stress incontinence frequency at the 28 gestational week in the no exercises group was 17.2%, and at the 35 gestational week of 47% and at 6 weeks after childbirth was of 47%, while in the exercises group was 0, 0 and 15%, respectively. Moreover there were statistically significant differences between both groups regarding the presence of urinary stress incontinence, that is, the group that realized exercises presented less incontinence at the 28 and 35 gestational weeks and at 6 weeks after childbirth. CONCLUSIONS: The results of this study concluded that the pelvic floor exercises during pregnancy and after childbirth prevent the urinary stress incontinence at this time. PMID- 15813474 TI - [Value of the morphological characteristics of polar body as a predictive factor of embryo development]. AB - OBJECTIVE: To determine the relation between polar body quality and perivitelline space with fertilization and segmentation rates and embryo quality. STUDY: Prospective and descriptive. MATERIAL AND METHODS: Sixty-one patients underwent the in vitro fertilization and embryo transfer program (IVF-ET). The intracytoplasmatic sperm-injection (ICSI) was applied for insemination. Controlled ovarian hyperstimulation was made with recombinant FSH. Oocytes were evaluated 4-6 hours after retrieval and polar body, perivitelline space, and pellucid zone were assessed too. Fertilization was evaluated 17 hours after ICSI, and segmentation rates were analyzed at 48 and 72 hours. The number of blastomeres and quality of each embryo was assessed prior transference. Statistical analysis was done with chi2 test. RESULTS: We evaluated 883 oocytes, of which 43.8% had a normal polar body, 54.9% had a normal perivitelline space, and 69% a normal pellucid zone; 57% of retrieved oocytes with normal polar body developed an adequate fertilization rate, and oocytes with abnormal polar body had fertilization in less than 20%. Embryo cleavage was adequate in oocytes with normal polar body, in the abnormal oocytes was less than 20%, and 79% of the oocytes with fragmented polar body did not developed in embryo. There were not significant differences in fertilization rates and segmentation when compared with the pellucid zone and perivitelline space. CONCLUSIONS: Oocyte quality has a great influence on fertilization and embryo development. One of the most important variables is the polar body quality, which correlates adequately with fertilization and embryo cleavage. PMID- 15813475 TI - [Estrogen role in the luteal phase support in in vitro fertilization with embryo transfer cycles]. AB - OBJECTIVE: To evaluate the effectiveness of estradiol administration for luteal phase support and to describe the progesterone and estradiol behavior in vitro fertilization-embryo transfer luteal phase. MATERIAL AND METHODS: Patients undergoing in vitro fertilization-embryo transfer with controlled ovarian hyperstimulation and using gonadotropin releasing hormone agonist. They were divided at random into two groups: group 1 would receive progesterone alone, and group 2 would take estrogen and progesterone. Serum concentrations of estradiol and progesterone were measured on days 7 and 14 post-embryo transfer. RESULTS: We examined 52 patients; 24 received progesterone alone and 28 took estrogen and progesterone. Significantly higher estradiol and progesterone concentrations on day 14 were found in pregnant women. It was not on day 7. A significant increment of estrogen was found in the estrogen and progesterone group. Progesterone did not increase significantly. Pregnancy rate was the same in both groups. CONCLUSIONS: For patients undergoing in vitro fertilization-embryo transfer, the addition of estradiol to the progesterone support regimen does not have beneficial effects in terms of pregnancy rate. On day 7 neither progesterone nor estradiol are good predictors of pregnancy. PMID- 15813476 TI - [Sexual frigidity. 1949]. PMID- 15813477 TI - [Acrosomal response of human sperm demonstrated with chlorotetracycline]. PMID- 15813478 TI - Minority faculty: another nursing shortage. PMID- 15813479 TI - Educational preparation of black nurses: a historical perspective. AB - To where minority nursing needs to proceed, the minority nursing community must understand where we have been. This historical perspective traces our roots through every level of nursing education. Parallels are drawn between minority nurse educational evolution and the historical events occurring in the greater society in the United States. PMID- 15813480 TI - Enhancing success in BSN nursing education for minority nurses. AB - Because of the critical need for minority nurses, especially nursing faculty, steps must be taken to recruit, and insure the success of minority students. Strategies for achieving graduation and NCLEX success among minority students attending a BSN program are outlined. Methods for enhancing nurse faculty skills for teaching diverse students are described. PMID- 15813481 TI - African American nursing faculty: where are they? AB - African American nursing faculty members are scarce in today's society. The National League for Nursing (NLN) developed a report of the faculty census survey of Registered Nurses (RN) and graduate programs in 2002. The report indicates that only 6.6% nursing faculty are African American compared to 91.0% Caucasian nursing faculty (NLN, 2003). Previous published literature explores reasons African Americans are not choosing academic careers, reasons for termination of employment, regions in the U.S. where African American faculty are employed, and strategies employed to recruit additional African American nursing faculty. PMID- 15813482 TI - Analysis of Disability Adjusted Life Years (DALYs) among arsenic victims: a cross sectional study on health economics perspective. AB - Arsenic contamination of ground water is a major public health problem in Bangladesh. It is estimated that more than 20 million people are potentially exposed to arsenic poisoning. This cross-sectional study was carried out in Haziganj Upazila under Chandpur district between September to December, 2001 with the objectives to assess the socioeconomic consequences and disease burden in terms of Disability Adjusted Life Years (DALYs). A total of 168 subjects suffering from arsenicosis were studied. Both age and disability weights were considered to calculate DALYs. Agricultural labour and housewives suffered more from the disease. A strong relationship (p<0.005) was found between duration of suffering and occupation of the subjects. Also, there was a strong relationship between age of onset and education of the study subjects (p<0.006). No deaths directly from arsenicosis were reported. It may be noticed that 47% of the patients would be living with disability for more than 51 years. A strong relationship exists (p<0.002) between educational level and Years Lived with Disability (YLDs). A total of 7930 YLDs were lost due to arsenicosis, which accounts for 1908 DALYs. PMID- 15813483 TI - Role of free to total prostate specific antigen ratio in serum in the diagnosis of prostatic enlargement. AB - This study was aimed to demonstrate the significance of free/total Prostate Specific Antigen (PSA) ratio in the diagnosis of prostatic enlargement. The study was carried out in a total number of 100 subjects, of which 58 had benign prostatic hyperplasia (BPH), 42 had carcinoma prostate and 50 were age matched control. The subjects were collected from Dhaka Medical College Hospital (DMCH) and Bangabandhu Sheikh Mujib Medical University (BSMMU). Venous blood (5ml) was drawn from each subject before digital rectal examination or pre-urethral manipulation for estimation of PSA. Final diagnosis was made by histopathological examination, specimen being obtained by transurethral resection of prostate (TURP), open prostatectomy and per rectal tru-cut biopsy with biopsy gun. The study showed a highly significant difference of serum PSA and free/total PSA ratio in differentiating BPH from carcinoma prostate. Free/total PSA ratio was highly significant in differentiating between BPH and carcinoma prostate (p<0.001) and carcinoma prostate and control (p<0.001). In the marginally elevated PSA, free/total PSA ratio reduced biopsy by 81-85% in BPH and carcinoma prostate respectively. The overall specificity, positive predictive value and efficiency of free/total PSA was much higher than that of total PSA in differentiating BPH from carcinoma prostate. PMID- 15813484 TI - Prevalence of type 2 diabetes in urban slums of Dhaka, Bangladesh. AB - This cross-sectional study was conducted to estimate the prevalence of type 2 diabetes along with its risk factors in urban slum population of Dhaka, Bangladesh. A random sample of 1555 slum dwellers of Dhaka city (age > or = 20 years) were included in the study. Capillary blood glucose levels, fasting and 2 h after 75g oral glucose load (for a selected subjects, n = 476), were measured. Height, weight, waist and hip circumferences, blood pressure and some other important socio-demographic information on age, sex, education, income, and occupation status were collected. The overall prevalence of type 2 diabetes was found to be 8.1 percent, and the prevalence for men and women were 7.7 percent and 8.5 percent respectively. Prevalence of diabetes was found to be lower following 2-h glucose values in the selected population compared to the FBG procedure. Age, sex, literacy and waist to hip ratio for men were found as significant risk factors following both fasting blood glucose and 2-h post glucose values adjusted for a number of confounding variables. Poor to moderate agreement was observed between fasting blood glucose and 2-h glucose (kappa 0.41, p < 0.001). The agreement was even poorer between impaired fasting glucose and impaired glucose tolerance. Poor agreement between FBG and 2-h BG may raise concern for the dependability of diagnostic procedures. Higher prevalence of type 2 diabetes in the urban slum may indicate an epidemiological transition due to fast urban migration and possibly urbanization. However, this issue needs further exploration. PMID- 15813485 TI - Interferon-gamma (IFN-gamma) response to different hepatitis B virus antigens in hepatitis B virus infection. AB - The IFN-gamma levels in serum and cultured supernatant of peripheral blood mononuclear cells (PBMCs) were compared after stimulation by HBsAg ad, HBsAg ay and HBcAg among 3 groups of subjects i.e. patients with acute HBV infection, patients with chronic HBV infection and subjects recovered from HBV infection. Uninfected vaccinated group was taken as control. Serum and PBMCs were obtained from 38 individuals between 18-50 years of age. PBMCs were separated from heparinised blood by Ficoll-Hypaque density gradient centrifugation technique and cultured in CO2 incubator after stimulation by HBV surface and core antigens. IFN gamma concentration was measured in serum and culture supernatant of PBMCs by an in-house ELISA technique. The mean serum IFN-gamma levels in acute, chronic, recovered and control groups were 88 pg/ml, 96.6 pg/ml, 155 pg/ml and 205 pg/ml respectively. On stimulation by HBsAg ad, IFN-gamma levels in cultured PBMCs of the above mentioned groups were 282.50 pg/ml, 307.45 pg/ml, 915.62 pg/ml and 511.67 pg/ml respectively, while in the same group on HBsAg ay stimulation, IFN gamma levels were 246.25 pg/ml, 374.70 pg/ml, 1040 pg/ml and 465.83 pg/ml respectively. On stimulation by HBcAg, the IFN-gamma levels were 875 pg/ml, 128.50 pg/ml, 905 pg/ml and 235.33 pg/ml respectively in the acute, chronic, recovered and control groups. When compared with serum, significantly higher levels of IFN-gamma in cultured supernatant of PBMCs were observed after stimulation by HBsAg ad and HBsAg ay subtype in cases of chronic (p<0.05) and recovered groups (p<0.01 and p<0.001 respectively). However, no statistically significant difference of IFN-gamma level was observed between serum and PBMCs amongst the acute and control groups when stimulated by either of the HBsAg subtypes or HBcAg. In the recovered group, IFN-gamma levels produced by PBMCs after stimulation by HBcAg were significantly higher than that of serum (p<0.01). The study concludes that on subsequent exposure, PBMCs of the recovered group produces higher levels of IFN-gamma in response to different hepatitis B antigens. This response perhaps is able to protect individuals who are unable to develop anti-HBs. PMID- 15813486 TI - Holmium laser urethrotomy for urethral stricture. AB - A prospective cross-sectional study was carried-out in the department of urology, Dhaka Medical College Hospital (DMCH) to evaluate the outcome of Laser urethrotomy for the treatment of urethral stricture. For this purpose, 30 male patients aged 15 to 60 years with short segment anterior urethral stricture (>2cm) were treated by HO:YAG Laser. The energy used for this purpose was 0.8 to 1.5 joules by LISA 80 Watt Holmium Laser machine. All patients were catheterized for less than 24 hours and were followed up for 6 to 12 months postoperatively by uroflowmetry and by retrograde with voiding cystourethrogram 3 monthly. The study revealed that out of 30 patients, 27(90%) showed good flow of urine (Qave>16.0 ml/sec) and adequate caliber urethra in retrograde urethrogram (RGU). Only 3(10%) patients showed narrow stream of urine (Qave<8.0 ml/sec) and recurrent stricture in RGU which were managed by optical internal urethrotomy (OIU) and clean intermittent self catheterization (CISC). The study showed satisfactory results in 90% cases with short term follow up. The study concludes that HO:YAG Laser urethrotomy for the treatment of short segment urethral stricture is highly effective. The study further reveals that the method is simple, safe and thus, it can be considered favorably as a new therapeutic option for the treatment of urethral stricture. However, long term follow up is necessary for making a final comment on this issue. PMID- 15813487 TI - On doing two things at once: temporal constraints on actions in language comprehension. AB - In two experiments, we investigated how text comprehension is influenced by the interaction between the properties of actions and the temporal relations specified by adverbs. Participants read short narratives describing a protagonist who performed two actions that involved similar sensorimotor systems (e.g., chopping wood and painting a fence) or different ones (e.g., whistling a melody and painting a fence). The actions were described as simultaneous or successive by means of the temporal adverbs while and after, respectively. Comprehension, both in Spanish and in English, was markedly impaired (longer reading times and lower subjective coherence) for sentences including the adverb while and actions involving the same sensorimotor system. However, when one of the same sensorimotor system actions was described as a mental plan (e.g., chopping wood and thinking of painting a fence), comprehension was equally easy with the adverbs while and after. These results are compatible with a revised version of the indexical hypothesis that specifies how comprehension is guided by syntax and embodied constraints within multiple noninteracting mental spaces. PMID- 15813488 TI - Character profiles and the activation of predictive inferences. AB - Most evidence indicates that with supportive context, predictive inferences do become activated. The present experiments were designed to investigate whether the characteristics of the protagonist can mitigate against activation of a predictive inference, even when the immediately preceding context supports it. Participants read passages containing a detailed description of characteristics of the protagonist. The characteristics were either consistent, inconsistent, or neutral with respect to a subsequent predictive inference. This character information was followed by a context that supported the predictive inference. Experiment 1 demonstrated that predictive inferences were activated, except when they conflicted with the characteristics of the protagonist. Experiment 2 demonstrated that when an inference was activated, it was also instantiated into the representation of the text. These results are discussed in terms of the memory-based view of text processing. PMID- 15813489 TI - Reducing structural-element salience on a source problem produces later success in analogical transfer: what role does source difficulty play? AB - Two experiments in reasoning by analogy were conducted to study the role of inducing source difficulty by reducing the salience of the source's structural elements. Three nonexclusive hypotheses were tested. According to the first, a difficult source problem improves analogical transfer because it increases the probability that the subject will notice the similarity between the source and the target. For example, errors made on both the source and the target can enhance the subject's awareness of the similarity between the two problems. According to the second hypothesis, a source that is difficult to solve is memorized better than an easier source. According to the third, source-problem difficulty affects the degree of abstractness in the representation of the solution elaborated by subjects. Experiment 1 showed that the higher frequency of spontaneous transfer between the source and the target when the source problem was difficult (Gick & McGarry, 1992) could be replicated in a cued-transfer situation. Experiment 2 showed that subjects given a difficult source, one in which the important element was not very salient, were better at categorizing isomorphic problems on the basis of structural features than were subjects given an easy source. The discussion deals with the implications of these results for the hypotheses tested and, more generally, for reasoning by analogy and education in general. PMID- 15813490 TI - Frequency of judgment as a context-like determinant of predictive judgments. AB - Several studies have shown that predictive and causal judgments vary depending on whether the question used to assess the relationship between events is presented after each piece of information or only after all the available information has been observed. This effect could be understood by assuming that in the two cases people perceive that the test question requires that different sets of evidence be taken into account. This hypothesis is tested in the present experiments through contextual manipulations that take place at the time of training and at the time of test. Our results show that people use this contextual information to infer which set of events should be considered when making their subjective assessments. The results are at odds with current theoretical approaches, but it is possible to develop mechanisms that would allow these models to account for the observed evidence. PMID- 15813491 TI - Dilution and confirmation of probability judgments based on nondiagnostic evidence. AB - Previous research has shown that probability judgments based on a mix of diagnostic and nondiagnostic information are less extreme than judgments based on the diagnostic information alone. Results of the present experiments suggest that this dilution effect holds only under a limited set of conditions. When judgments based on a mix of diagnostic and nondiagnostic information are compared with separately elicited judgments based on the diagnostic information alone, the dilution effect is consistently observed. When judgments based on the diagnostic evidence are revised in light of additional, nondiagnostic evidence, by contrast, the dilution effect is eliminated or even reversed (yielding a confirmation effect) depending on the type of nondiagnostic evidence under evaluation. PMID- 15813492 TI - Blocking by word frequency and neighborhood density in visual word recognition: a task-specific response criteria account. AB - Effects of blocking words by frequency class (high vs. low) and neighborhood density (high vs. low) were examined in two experiments using progressive demasking and lexical decision tasks. The aim was to examine the predictions of a task-specific response criteria account of list-blocking effects. Distinct patterns of blocking effects were obtained in the two tasks. In the progressive demasking task, a pure-list disadvantage was obtained to low frequency-high density words, whereas high frequency-low density produced a trend toward a pure list advantage. In lexical decision, high-frequency words showed a pure-list advantage that was strongest in high-density words, whereas low frequency-low density words produced a trend toward a pure-list disadvantage. A simulation study implementing task-specific response criteria within the framework of the multiple read-out model provided an accurate description of the blocking effects obtained in the experiments. It is argued that adjustments of task-specific response criteria determine changes in list-blocking effects across different tasks. PMID- 15813493 TI - On the process of recognizing inverted words: does it rely only on orientation invariant cues? AB - Following a demonstration by Parks (1983) of failure to notice the reflection of a letter of an inverted word, two experiments were conducted to test a hypothesis about the process of recognizing inverted words that is termed here invariant cues only (ICO)-a letter-by-letter identification process based only on orientation-invariant letter features. In Experiment 1, subjects were presented with whole strings-words and nonwords, either upright or inverted-in which either all the letters were normal or one of the letters was reflected, and they were asked to make lexical decisions. In Experiment 2, subjects made a reflection judgment about an upright or inverted letter within a string immediately after they had been presented with the other, nonreflected string letters, again either upright or inverted. The results do not support the ICO hypothesis: Lexical decisions were greatly affected by the reflection of a letter in upright and inverted stimuli alike. Reflection judgments were considerably facilitated by word context in the upright and the inverted modes alike. The results are accommodated better by the notion that recognition of disoriented words requires some correction used to restore orientation-sensitive features. PMID- 15813494 TI - Emotional intensity predicts autobiographical memory experience. AB - College students generated autobiographical memories from distinct emotional categories that varied in valence (positive vs. negative) and intensity (high vs. low). They then rated various perceptual, cognitive, and emotional properties for each memory. The distribution of these emotional memories favored a vector model over a circumplex model. For memories of all specific emotions, intensity accounted for significantly more variance in autobiographical memory characteristics than did valence or age of the memory. In two additional experiments, we examined multiple memories of emotions of high intensity and positive or negative valence and of positive valence and high or low intensity. Intensity was a more consistent predictor of autobiographical memory properties than was valence or the age of the memory in these experiments as well. The general effects of emotion on autobiographical memory properties are due primarily to intensity differences in emotional experience, not to benefits or detriments associated with a specific valence. PMID- 15813495 TI - Construct validity and age sensitivity of prospective memory. AB - We administered four prospective memory tasks to 330 adults between 18 and 89 years of age to investigate the relationship among the measures of performance in the four tasks, as well as the relationship of the prospective memory measures to age, other cognitive abilities, and noncognitive factors. The four prospective memory variables were found to exhibit both convergent and discriminant validity, indicating that prospective memory ability appears to represent a distinct dimension of individual differences. The prospective memory construct was significantly related to other cognitive abilities, such as executive functioning, fluid intelligence, episodic memory, and perceptual speed, but it was only weakly related to self-ratings of (primarily retrospective) memory and to personality traits. Although a substantial proportion of the age-related variance on the prospective memory construct was shared with other cognitive abilities, we also found some evidence of unique, statistically independent, age related influences on prospective memory. PMID- 15813496 TI - Memory for thematically arousing events. AB - Many studies have indicated that emotional arousal improves memory for the center, or gist, of an event but undermines memory for the event's periphery. However, all of these studies have elicited emotion by showing participants some salient visual stimulus intended to arouse them (e.g., the sight of a wound). This stimulus may have served as an attention magnet, and this, not the arousal, may have been the cause of the observed narrowing of memory. In this article, we examine how participants remember events that involve thematically induced arousal, arousal produced by empathy, rather than by a visual emotional stimulus. The data show that emotionality improves memory for all aspects of these events, with no memory narrowing. PMID- 15813497 TI - Selective sex differences in declarative memory. AB - Sex invariance of a six-factor, higher order model of declarative memory (two second-order factors: episodic and semantic memory; and four first-order factors: recall, recognition, fluency, and knowledge) was established for 1,796 participants (35-85 years). Metric invariance of first- and second-order factor loadings across sex was demonstrated. At the second-order level, a female advantage was observed for both episodic and semantic memory. At the first-order level, sex differences in episodic memory were apparent for both recall and recognition, whereas the differences in semantic memory were driven by a female superiority in fluency. Additional tests of sex differences in three age groups (35-50, 55-65, and 70-85 years of age) indicated that the female superiority in declarative memory diminished with advancing age. The factor-specific sex differences are discussed in relation to sex differences in hippocampal function. PMID- 15813498 TI - Depth rotation and mirror-image reflection reduce affective preference as well as recognition memory for pictures of novel objects. AB - In two experiments, the identification of novel 3-D objects was worse for depth rotated and mirror-reflected views, compared with the study view in an implicit affective preference memory task, as well as in an explicit recognition memory task. In Experiment 1, recognition was worse and preference was lower when depth rotated views of an object were paired with an unstudied object relative to trials when the study view of that object was shown. There was a similar trend for mirror-reflected views. In Experiment 2, the study view of an object was both recognized and preferred above chance when it was paired with either depth rotated or mirror-reflected views of that object. These results suggest that view sensitive representations of objects mediate performance in implicit, as well as explicit, memory tasks. The findings do not support the claim that separate episodic and structural description representations underlie performance in implicit and explicit memory tasks, respectively. PMID- 15813499 TI - Effects of repetition and response deadline on associative recognition in young and older adults. AB - The present study examined the joint effects of repetition and response deadline on associative recognition in older adults. Young and older adults studied lists of unrelated word pairs, half presented once (weak pairs) and half presented four times (strong pairs). Test lists contained old (intact) pairs, pairs consisting of old words that had been studied with other partners (rearranged lures), and unstudied pairs (new lures), and participants were asked to respond "old" only to intact pairs. In Experiment 1, participants were tested with both short and long deadlines. In Experiment 2, the tests were unpaced. In both experiments, repetition increased hit rates for young and older adults. Young adults tested with a long deadline showed reduced (Experiment 1) or invariant (Experiment 2) false alarms to rearranged lures when word pairs were studied more often. Young adults tested with a short deadline and older adults tested under all conditions had increased false alarm rates forstrong rearranged pairs. Implications of these results for theories of associative recognition and cognitive aging are explored. PMID- 15813500 TI - The cognitive processing of film and musical soundtracks. AB - Previous research has demonstrated that musical soundtracks can influence the interpretation, emotional impact, and remembering of film information. The intent here was to examine how music is encoded into the cognitive system and subsequently represented relative to its accompanying visual action. In Experiment 1, participants viewed a set of music/film clips that were either congruent or incongruent in their emotional affects. Selective attending was also systematically manipulated by instructing viewers to attend to and remember the music, film, or both in tandem. The results from tune recognition, film recall, and paired discrimination tasks collectively revealed that mood-congruent pairs lead to a joint encoding of music/film information as well as an integrated memory code. Incongruent pairs, on the other hand, result in an independent encoding in which a given dimension, music or film, is only remembered well if it was selectively attended to at the time of encoding. Experiment 2 extended these findings by showing that tunes from mood-congruent pairs are better recognized when cued by their original scenes, while those from incongruent pairs are better remembered in the absence of scene information. These findings both support and extend the "Congruence Associationist Model" (A. J. Cohen, 2001), which addresses those cognitive mechanisms involved in the processing of music/film information. PMID- 15813501 TI - Interpreting the parameters of the diffusion model: an empirical validation. AB - The diffusion model (Ratcliff, 1978) allows for the statistical separation of different components of a speeded binary decision process (decision threshold, bias, information uptake, and motor response). These components are represented by different parameters of the model. Two experiments were conducted to test the interpretational validity of the parameters. Using a color discrimination task, we investigated whether experimental manipulations of specific aspects of the decision process had specific effects on the corresponding parameters in a diffusion model data analysis (see Ratcliff, 2002; Ratcliff & Rouder, 1998; Ratcliff, Thapar, & McKoon, 2001, 2003). In support of the model, we found that (1) decision thresholds were higher when we induced accuracy motivation, (2) drift rates (i.e., information uptake) were lower when stimuli were harder to discriminate, (3) the motor components were increased when a more difficult form of response was required, and (4) the process was biased toward rewarded responses. PMID- 15813502 TI - Differential gene expression in liposarcoma, lipoma, and adipose tissue. AB - Malignant transformation is thought to be associated with changes in the expression of a number of genes, and this alteration in gene expression is felt to be critical to the development of the malignant phenotype. Sarcomas represent a diverse group of tumors derived from cells of mesenchymal origin. Marked heterogeneity exists in the biological behavior of sarcomas, even within histologic subtypes of sarcomas. In an effort to better understand the biology of liposarcomas, gene expression in normal adipose tissue, lipomas, and liposarcomas was examined using the Affymetrix microarray technology. Differences in gene expression were quantified as the fold change in gene expression among the sample sets. Differences in gene expression among normal adipose tissue, lipomas, and liposarcomas were observed. In addition, genes expressed uniquely in liposarcoma among these and 18 other tissue sample sets were identified. Gene sets were devised that allowed the separation of liposarcomas from other samples, and most normal adipose tissue from most lipomas using the Eisen clustering software "Cluster." We conclude that differences in gene expression can be identified among different tumors derived from the adipocyte series. Such differences in gene expression may help differentiate among subtypes of sarcomas, and may also yield clues to the pathophysiology of this heterogeneous group of tumors. PMID- 15813503 TI - Tc-99m labelled anti CD 66 antigranulocyte antibody imaging for assessment of bone marrow reserve. AB - Patients with metastatic disease to the bone marrow are at risk of significant hematologic toxicity, if they undergo myelotoxic chemotherapy. Tc-99m labelled Anti CD 66 monoclonal antibody imaging is a useful, noninvasive approach to the assessment of bone marrow reserve under such circumstances. PMID- 15813504 TI - Gemcitabine and cisplatin in advanced nasopharyngeal carcinoma: a pilot study. AB - A pilot study was performed to evaluate the efficacy and safety of gemcitabine and cisplatin combination in the treatment of patients with metastatic nasopharyngeal carcinoma (NPC). Eligible patients were those with metastatic NPC who had been treated with radiotherapy and cisplatin plus 5-fluorouracil chemotherapy. Cisplatin was given intravenously at the fixed dose of 30 mg/m2 on days 1-3. Gemcitabine was intravenously administered over 30 min infusion with the dose escalated from 800 to 1200 mg/m2 on days 1 and 8. The 3-week schedule defined a cycle of treatment. Fifteen patients were enrolled and assessed for the worst toxicities. For a total of 83 cycles, Grade 3-4 toxicity was 46.7 % for neutropenia, 40.0 % for thrombocytopenia, and 20.0% for anemia. Grade 3 nonhematologic toxicity was 13.3%. Fourteen patients were assessable for response. The overall response rate was 92.9%, with complete response in three patients (21.4%). Median survival was 10.2 months. Seven patients had lived more than one year, and two patients had lived more than 2 years. The recommended dose of gemcitabine was 1000 mg/m2 on days 1 and 8 in each cycle. In conclusion, the present combination is well tolerated and highly active in the treatment of advanced NPC patients. PMID- 15813505 TI - Tumor motion control in the treatment of non small cell lung cancer. AB - Tumor motion due to respiration during radiation therapy for non-small cell lung cancer is a significant problem. This article reports on two techniques used to control tumor motion: respiratory gating and the deep inspiration breath hold technique. This technique was implemented in 40 patients without significant difficulties and there are encouraging clinical outcomes. PMID- 15813506 TI - Response to temozolomide in second-line treatment of recurrent nonsmall cell lung carcinoma: case report. AB - Almost all patients with stage IV nonsmall cell lung cancer (NSCLC) who show an initial response to chemotherapy will eventually relapse. For patients with a good performance status at relapse, second-line chemotherapy is a standard treatment option. A case of recurrent NSCLC described herein profiles a patient previously treated with cisplatin, irinotecan, and radical thoracic radiation for stage IIIb NSCLC. The patient showed a complete response to first-line therapy that lasted for approximately 17 months. When new adenopathy was discovered, therapy with the oral chemotherapeutic agent temozolomide was initiated. After 2 treatment cycles, a near complete response was obtained and she remained free from disease progression during all subsequent treatment cycles with temozolomide. The patient remained free from disease progression for a total of 15 months when she was treated for dehydration and a computed tomography (CT) scan showed new small bilateral pleural effusions and enlarging subcarinal, right hilar, and left infrahilar lymph nodes. The patient died 21 months after her first treatment cycle with temozolomide, most likely because of progressive disease. PMID- 15813507 TI - Benefit of temozolomide compared to procarbazine in treatment of glioblastoma multiforme at first relapse: effect on neurological functioning, performance status, and health related quality of life. AB - Since high-grade malignant gliomas can seldom be treated curatively, the main aim of first line therapy is to improve progression free survival (PFS), to reduce morbidity, and to preserve, if not restore neurological functions and the capacity to perform daily activities. Focusing on a single clinical efficacy parameter in clinical trials may provide a potentially biased result, as for patients the overall result of treatment entails a more complex picture of weighing and balancing gains and losses on different outcome measures. In this paper we address different clinical outcomes measures separately and we illustrate the value of multiple outcome measures using the results of a recent clinical trial comparing temozolomide with procarbazine in the treatment of Glioblastoma Multiforme. Compared with procarbazine, temozolomide not only prolonged PFS, but also maintained neurological functioning and performance status for a longer period of time, and also improved health-related quality of life (HRQL). All these statistically significant outcomes demonstrate a remarkable consistency. In addition, temozolomide showed a trend of extending overall survival over procarbazine. PMID- 15813508 TI - Leptomeningeal metastasis. AB - Leptomeningeal metastasis is a common complication of cancer. Often the diagnosis can be difficult, but early diagnosis and aggressive treatment can prevent irreversible neurologic deficits. Diagnosis is usually established by the demonstration of malignant cells in the cerebrospinal fluid (CSF) or by the presence of enhancing tumor nodules on cranial or spinal MRI. Treatment may require focal radiotherapy to symptomatic sites accompanied by chemotherapy. Intra-CSF chemotherapy requires normal CSF flow dynamics and is typically limited to the use of methotrexate, cytarabine or thiotepa. Intrathecal chemotherapy is rarely efficacious, except for hematopoietic neoplasms and, to a lesser extent, breast cancer. Increasingly, systemic chemotherapy is recognized as efficacious in the treatment of leptomeningeal metastasis in part because it has the ability to penetrate into bulky disease seen on neuroimaging. The optimum choice of therapy depends upon a thorough assessment of the neurologic extent of disease, which should include complete neuraxis imaging. Patients with extensive bulky disease may be best treated with systemic chemotherapy, whereas those with a positive CSF cytology but negative imaging may be treated with intrathecal chemotherapy and spared the systemic toxicity of intravenous drug. Despite vigorous therapy, many patients do poorly and the median survival is only about four months. Nevertheless, some patients, particularly those with leukemia, lymphoma, and breast cancer, respond and a substantial minority will be alive one to two years after diagnosis. PMID- 15813509 TI - Strategies for the prevention and treatment of peritoneal carcinomatosis from gastrointestinal cancer. AB - BACKGROUND: Peritoneal surface malignancy can result from full thickness invasion of gastrointestinal cancer through the bowel wall or from dissemination of cancer cells from the trauma of cancer surgery. In the past, this clinical situation was treated only with palliative intent. METHODS: An aggressive approach to peritoneal surface malignancy involves peritonectomy procedures, perioperative intraperitoneal chemotherapy and knowledgeable patient selection. The clinical assessments necessary for valid clinical judgements include the cancer histopathology (invasive vs. expansive progression), the preoperative abdominal and pelvic CT, the peritoneal cancer index and the completeness of cytoreduction score. Proper patient selection is mandatory for optimizing the results of treatment. RESULTS: In a series of phase II studies, appendiceal tumors with peritoneal seeding became the paradigm for success with an 85% long-term survival in selected patients. Carcinomatosis from colon cancer had an overall 5-year survival of 50% with selected patients. In all malignancies, early aggressive treatment of minimal peritoneal surface dissemination showed the greatest benefit. CONCLUSIONS: Oncologists must accept responsibility for knowledgeable management of peritoneal surface dissemination of cancer because a curative approach has been demonstrated in both phase II studies and phase III studies. All historical controls show 0% long-term survival. Surgical interventions combined with perioperative intraperitoneal chemotherapy in diseases where peritoneal surface spread occurs must be considered a treatment option. PMID- 15813510 TI - Preclinical and clinical experience with fulvestrant (Faslodex) in postmenopausal women with hormone receptor-positive advanced breast cancer. AB - Fulvestrant (Faslodex) is a new type of estrogen receptor (ER) antagonist that binds, blocks, and degrades the ER, leading to reduced expression of the progesterone receptor (PgR). Unlike the selective ER modulator tamoxifen, fulvestrant is devoid of any known agonist activity. Fulvestrant has a steroidal structure that competitively binds to the ER with an affinity much greater than that of tamoxifen. After binding to fulvestrant, degradation of the ER is accelerated, ultimately resulting in a reduction in cellular ER. Immunohistochemical studies have demonstrated that a single intramuscular injection of fulvestrant results in a dose-dependent decrease in ER and PgR indices and in Ki-67 expression. At the approved 250 mg dose, the decrease in receptor positivity with fulvestrant treatment was greater than that with tamoxifen. Phase III clinical trials have demonstrated the clinical benefit of fulvestrant in the endocrine treatment of breast cancer. Among patients who progressed during adjuvant or first-line endocrine therapy, fulvestrant was at least as effective as anastrozole as second-line treatment for the primary endpoints of objective response and time to progression, and was well tolerated. After a median follow-up of 24.5 months, a combined survival analysis from two Phase III studies has shown that fulvestrant may be considered similar to anastrozole for time to death (27.4 months versus 27.7 months, respectively). For the first-line therapy of advanced breast cancer in postmenopausal women, fulvestrant was shown to be active and well tolerated in a trial that compared fulvestrant 250 mg once monthly and tamoxifen 20 mg once daily. Treatment with fulvestrant has also demonstrated clinical efficacy among patients who progressed following treatment with tamoxifen followed by nonsteroidal aromatase inhibitors. The efficacy of fulvestrant in sequential endocrine therapy and in combination with other agents appears promising and active investigations are ongoing to explore the clinical potential of this novel antiestrogen. PMID- 15813511 TI - The epidemiology of cancer pain. AB - We identified 28 epidemiological surveys by applying a sensitive search in Medline and CancerLit databases, supplemented by hand searches. Only two surveys enrolled more than 10,000 patients with cancer. The remaining were hospital or clinic-based surveys of at most a few hundred patients. Fourteen surveys were conducted in the United States. The majority of the remaining studies were conducted in Europe (Finland, France, Germany, UK/Ireland). No single survey identified a prevalence of any type of pain below 14%. The prevalence of pain reported in these surveys varies with the specific type of pain (e.g., breakthrough pain) and/or population studied. Based on these surveys an aggregate statement could not be deduced regarding the correlation between the occurrence of pain and patient factors, disease characteristics, the setting in which care is provided (e.g., primary care or specialized oncology or pain treatment clinics), or specific treatments directed towards the underlying disease and its associated pain. However, these surveys suggest that a significant number of patients with cancer worldwide will, during the course of their disease, experience pain that requires medical and/or other treatment. PMID- 15813512 TI - Complete remission with anti-CD20 therapy for unicentric, non-HIV-associated, hyaline-vascular type, Castleman's disease. PMID- 15813513 TI - Improved interferometric detection of scattered light with a 4f imaging system. AB - We analyze the performance of three imaging systems to detect near-forward scattered light interferometrically by using a Mach-Zehnder geometry. The alignment of each system is demonstrated by measurement of the heterodyne efficiency and correlation of the angular width and field 1/e radius measurements of the sample beam. Measurements of angular-scattering data demonstrate the range of angles over which each system is effective. Of the three systems analyzed, the 4f imaging system is determined to be most effective, because it accurately reproduces both the phase and the amplitude of the scattered field at the detector. PMID- 15813514 TI - In vivo fiber-optic confocal reflectance microscope with an injection-molded plastic miniature objective lens. AB - For in vivo optical diagnostic technologies to be distributed to the developed and developing worlds, optical imaging systems must be constructed of inexpensive components. We present a fiber-optic confocal reflectance microscope with a cost effective injection-molded plastic miniature objective lens for in vivo imaging of human tissues in near real time. The measured lateral resolution is less than 2.2 microm, and the measured axial resolution is 10 microm. Confocal images of ex vivo cervical tissue biopsies and in vivo human lip taken at 15 frames/s demonstrate the microscope's capability of imaging cell morphology and tissue architecture. PMID- 15813515 TI - Fourier-domain holographic optical coherence imaging of tumor spheroids and mouse eye. AB - Fourier-domain holography (FDH) has several advantages over image-domain holography for optical coherence imaging of tissue. Writing the hologram in the Fourier plane significantly reduces background arising from reference light scattered from the photorefractive holographic film. The ability to use FDH is enhanced by the use of a diffuse target, such as scattering tissue, rather than specular targets, because the broader angular distribution from diffuse targets is transformed into a relatively uniform distribution in the Fourier plane. We demonstrate significantly improved performance for Fourier-domain optical coherence imaging on rat osteogenic sarcoma tumor spheroids and mouse eye. The sensitivity is documented at -95 dB. PMID- 15813516 TI - Time-domain optical coherence tomography with digital holographic microscopy. AB - We show that digital holography can be combined easily with optical coherence tomography approach. Varying the reference path length is the means used to acquire a series of holograms at different depths, providing after reconstruction images of slices at different depths in the specimen thanks to the short coherence length of light source. A metallic object, covered by a 150-microm thick onion cell, is imaged with high resolution. Applications in ophthalmology are shown: structures of the anterior eye, the cornea, and the iris, are studied on enucleated porcine eyes. Tomographic images of the iris border close to the pupil were obtained 165 microm underneath the eye surface. PMID- 15813517 TI - Comparative performance analysis of time-frequency distributions for spectroscopic optical coherence tomography. AB - The analysis of spectroscopic optical coherence tomography (SOCT) signals suffers the trade-off between time resolution and frequency resolution. Various joint time-frequency distributions (TFDs) can optimize this trade-off. Synthesized signals were generated and experimentally acquired data were obtained to compare and validate several different TFDs under different SOCT imaging schemes. Specific criteria were designed to quantify the TFD performance. We found that different SOCT imaging schemes require different optimal TFDs. Cohen's class TFDs generate the most compact time-frequency (TF) analysis, while linear TFDs offer the most reliable TF analysis. In both cases, if some prior information is known, model-based TF analysis can improve the performance. PMID- 15813518 TI - Determination of optimal exposure time for imaging of blood flow changes with laser speckle contrast imaging. AB - Laser speckle contrast imaging is becoming an established method for full-field imaging of cerebral blood flow dynamics in animal models. The sensitivity and noise in the measurement of blood flow changes depend on the camera exposure time. The relation among sensitivity, noise, and camera exposure time was investigated experimentally by imaging the speckle contrast changes in the brain after electrical forepaw stimulation in rats. The sensitivity to relative changes in speckle contrast was found to increase at longer exposure times and to reach a plateau for exposure times greater than approximately 2 ms. However, the speckle contrast noise also increases with exposure time and thus the contrast-to-noise ratio was found to peak at an exposure time of approximately 5 ms. Our results suggests that approximately 5 ms is an optimal exposure time for imaging of stimulus-induced changes in cerebral blood flow in rodents. PMID- 15813519 TI - Wavelength dependence of the apparent diameter of retinal blood vessels. AB - Imaging of retinal blood vessels may assist in the diagnosis and monitoring of diseases such as glaucoma, diabetic retinopathy, and hypertension. However, close examination reveals that the contrast and apparent diameter of vessels are dependent on the wavelength of the illuminating light. In this study multispectral images of large arteries and veins within enucleated swine eyes are obtained with a modified fundus camera by use of intravitreal illumination. The diameters of selected vessels are measured as a function of wavelength by cross sectional analysis. A fixed scale with spectrally independent dimension is placed above the retina to isolate the chromatic effects of the imaging system and eye. Significant apparent differences between arterial and venous diameters are found, with larger diameters observed at shorter wavelengths. These differences are due primarily to spectral absorption in the cylindrical blood column. PMID- 15813520 TI - Measurement of particle-size distribution and concentration in heterogeneous turbid media with multispectral diffuse optical tomography. AB - We present a method that is capable of extracting particle-size distribution (PSD) and concentration in heterogeneous turbid media by use of multispectral diffuse optical tomography (MSDOT). After the spectroscopic scattering images of the heterogeneous turbid media are obtained with MSDOT, the morphologic information of particles in the heterogeneities is recovered with an iterative regularized reconstruction algorithm based on Mie scattering theory when a particular form of PSD is assumed (Gaussian distribution is used in this study). The method described is tested and evaluated with both simulated and experimental data. The simulations are intended to test the sensitivity of the overall approach to noise effect. A series of phantom experiments are conducted with our newly developed ten-wavelength MSDOT system. Polystyrene microsphere suspensions contain particles of varying size from 2 to 6 microm as targets are embedded in a scattering background medium in these experiments. To achieve optimized results from experimental data, we developed a data preprocessing method for MSDOT as well as a scheme for calibrating scattering spectra. The results from both simulations and experiments show that the particle mean size and concentration can be reconstructed with acceptable accuracy, whereas the recovery of the standard deviation is sensitive to noise effect and can be as large as 86% from the experimental data. PMID- 15813521 TI - Experimental measurement of the refractive index of biological tissues by total internal reflection. AB - We discuss the refractive-index measurement of biological tissues by total internal reflection. The methodology of the measurement is illuminated comprehensively, and an experimental setup, combined with a data processing program, is developed correspondingly. Refractive indices of typical tissue samples are measured by use of the developed methodology. The agreement of our measurements with the reported results shows the validity of our scheme, which has the potential for being a simple, quick, and low-cost practical means for determining the refractive index of a turbid medium. Moreover, an empirical formula for evaluating the refractive index of Intralipid suspensions with different concentrations is also presented according to experimental measurements. PMID- 15813522 TI - Optical noninvasive monitoring of skin blood pulsations. AB - Time-resolved detection and analysis of skin backscattered optical signals (remission photoplethysmography or PPG) provide rich information on skin blood volume pulsations and can serve for reliable cardiovascular assessment. Single- and multiple-channel PPG concepts are discussed. Simultaneous data flow from several locations on the human body allows us to study heartbeat pulse-wave propagation in real time and to evaluate vascular resistance. Portable single-, dual-, and four-channel PPG monitoring devices with special software have been designed for real-time data acquisition and processing. The prototype devices have been clinically studied, and their potential for monitoring heart arrhythmias, drug-efficiency tests, steady-state cardiovascular assessment, body fitness control, and express diagnostics of the arterial occlusions has been confirmed. PMID- 15813523 TI - Spectrally constrained chromophore and scattering near-infrared tomography provides quantitative and robust reconstruction. AB - A multispectral direct chromophore and scattering reconstruction technique has been implemented for near-infrared frequency-domain tomography in recovering images of total hemoglobin, oxygen saturation, water, and scatter parameters. The method applies the spectral constraint of the chromophores and scattering spectra directly in the reconstruction algorithm, thereby reducing the parameter space of the inversion process. This new method was validated by use of simulated and experimental data, and results show better robustness and stability in the presence of higher levels of noise. The method suppresses artifacts, especially those significant in water and scatter power images, and reduces cross talk between chromophore and scatter parameters. Variation in scattering was followed by this spectral approach successfully in experimental data from 90-mm-diameter cylindrical phantoms, and results show linear variation in scatter amplitude and reduced scattering coefficient (micro(s)'), with total hemoglobin, oxygen saturation, and water remaining constant and quantitatively accurate. Similar experiments were carried out for varying oxygen saturation and total hemoglobin. Accurate quantification was obtained with a mean error of 7.7% for oxygen saturation and 6.2% for total hemoglobin, with minimal cross talk between different parameters. PMID- 15813524 TI - Effects of refractive index on near-infrared tomography of the breast. AB - Near infrared (NIR) optical tomography is an imaging technique in which internal images of optical properties are reconstructed with the boundary measurements of light propagation through the medium. Recent advances in instrumentation and theory have led to the use of this method for the detection and characterization of tumors within the female breast tissue. Most image reconstruction approaches have used the diffusion approximation and have assumed that the refractive index of the breast is constant, with a bulk value of approximately 1.4. We have applied a previously reported modified diffusion approximation, in which the refractive index for different tissues can be modeled. The model was used to generate NIR data from a realistic breast geometry containing a localized anomaly. Using this simulated data, we have reconstructed optical images, both with and without correct knowledge of the refractive-index distribution to show that the modified diffusion approximation can accurately recover the anomaly given a priori knowledge of refractive index. But using a reconstruction algorithm without the use of correct a priori information regarding the refractive-index distribution is shown as recovering the anomaly but with a degraded quality, depending on the degree of refractive index mismatch. The results suggest that provided the refractive index of breast tissue is approximately 1.3-1.4, their exclusion will have minimal effect on the reconstructed images. PMID- 15813525 TI - Computational calibration method for optical tomography. AB - We propose a computational calibration method for optical tomography. The model of the calibration scheme is based on the rotation symmetry of source and detector positions in the measurement setup. The relative amplitude losses and phase shifts at the optic fibers are modeled by complex-valued coupling coefficients. The coupling coefficients can be estimated when optical tomography data from a homogeneous and isotropic object are given. Once these coupling coefficients have been estimated, any data measured with the same measurement setup can be corrected for the relative variation in the data due to source and detector losses. The final calibration of the data for the source and detector losses and the source calibration between the data and the forward model are obtained as part of the initial estimation for reconstruction. The calibration method was tested with simulations and measurements. The results show that the coupling coefficients of the sources and detectors can be estimated with good accuracy. Furthermore, the results show that the method can significantly improve the quality of reconstructed images. PMID- 15813526 TI - Monitoring recovery after laser surgery of the breast with optical tomography: a case study. AB - Results are presented of a study to monitor the changes in the optical properties of breast tissue over a 12-month period after interstitial laser photocoagulation treatment of a fibroadenoma. The study involved generating cross-sectional images of the breast with a multichannel time-resolved imaging system and a nonlinear image reconstruction algorithm. Images of the internal absorbing and scattering properties revealed the expected initial inflammatory response, followed by the development of low-scattering cysts consistent with corresponding ultrasound examinations. Although results indicate that purely qualitative images can potentially provide clinically valuable data, means of enhancing diagnostic information by overcoming present limitations of the approach are discussed. PMID- 15813527 TI - Time-resolved diffuse optical tomographic imaging for the provision of both anatomical and functional information about biological tissue. AB - We present in vivo images of near-infrared (NIR) diffuse optical tomography (DOT) of human lower legs and forearm to validate the dual functions of a time-resolved (TR) NIR DOT in clinical diagnosis, i.e., to provide anatomical and functional information simultaneously. The NIR DOT system is composed of time-correlated single-photon-counting channels, and the image reconstruction algorithm is based on the modified generalized pulsed spectral technique, which effectively incorporates the TR data with reasonable computation time. The reconstructed scattering images of both the lower legs and the forearm revealed their anatomies, in which the bones were clearly distinguished from the muscles. In the absorption images, some of the blood vessels were observable. In the functional imaging, a subject was requested to do handgripping exercise to stimulate physiological changes in the forearm tissue. The images of oxyhemoglobin, deoxyhemoglobin, and total hemoglobin concentration changes in the forearm were obtained from the differential images of the absorption at three wavelengths between the exercise and the rest states, which were reconstructed with a differential imaging scheme. These images showed increases in both blood volume and oxyhemoglobin concentration in the arteries and simultaneously showed hypoxia in the corresponding muscles. All the results have demonstrated the capability of TR NIR DOT by reconstruction of the absolute images of the scattering and the absorption with a high spatial resolution that finally provided both the anatomical and functional information inside bulky biological tissues. PMID- 15813528 TI - Recovery of hemoglobin oxygen saturation and intrinsic fluorescence with a forward-adjoint model. AB - We present two forward-adjoint models for recovering intrinsic fluorescence spectra and hemoglobin oxygen saturation of turbid samples. The first fits measured diffuse reflectance spectra to obtain the absorption and scattering spectra of the medium, and these are then used to correct distortions imposed on the fluorescence spectrum by absorption and scattering. The second fits only the measured fluorescence spectrum to determine simultaneously the amplitudes of absorption and fluorescence basis spectra and scattering parameters. Both methods are validated with Monte Carlo simulations and experimentally in scattering phantoms containing nicotinamide adenine dinucleotide and human erythrocytes. Preliminary measurements from murine tumors in vivo are presented. PMID- 15813529 TI - Fluorescence spectra provide information on the depth of fluorescent lesions in tissue. AB - The fluorescence spectrum measured from a fluorophore in tissue is affected by the absorption and scattering properties of the tissue, as well as by the measurement geometry. We analyze this effect with Monte Carlo simulations and by measurements on phantoms. The spectral changes can be used to estimate the depth of a fluorescent lesion embedded in the tissue by measurement of the fluorescence signal in different wavelength bands. By taking the ratio between the signals at two wavelengths, we show that it is possible to determine the depth of the lesion. Simulations were performed and validated by measurements on a phantom in the wavelength range 815-930 nm. The depth of a fluorescing layer could be determined with 0.6-mm accuracy down to at least a depth of 10 mm. Monte Carlo simulations were also performed for different tissue types of various composition. The results indicate that depth estimation of a lesion should be possible with 2-3-mm accuracy, with no assumptions made about the optical properties, for a wide range of tissues. PMID- 15813530 TI - Detection of cortical activation with time-resolved diffuse optical methods. AB - Simulations based on diffusion theory that use a finite-element method and rely on an magnetic resonance imaging head model suggest that time-resolved diffuse optical techniques could provide information about the depth at which variations in perfusion take place and improve the detection of cortical activation. Experimental investigations were performed with sequentially driven picosecond laser diodes and an eight-channel time-correlated single-photon-counting detection system. The experimental results obtained for activation in the motor cortex, and for the Valsalva maneuver, confirm our assumptions and are in good agreement with the simulated data. PMID- 15813531 TI - Optimal linear inverse solution with multiple priors in diffuse optical tomography. AB - A general framework for incorporating single and multiple priors in diffuse optical tomography is described. We explore the use of this framework for simultaneously utilizing spatial and spectral priors in the context of imaging breast cancer. The utilization of magnetic resonance images of water and lipid content as a statistical spatial prior for the diffuse optical image reconstructions is also discussed. Simulations are performed to demonstrate the significant improvement in image quality afforded by combining spatial and spectral priors. PMID- 15813532 TI - Simulation study of magnetic resonance imaging-guided cortically constrained diffuse optical tomography of human brain function. AB - Diffuse optical imaging can measure brain activity noninvasively in humans through the scalp and skull by measuring the light intensity modulation arising from localized-activity-induced absorption changes within the cortex. Spatial resolution and localization accuracy are currently limited by measurement geometry to approximately 3 cm in the plane parallel to the scalp. Depth resolution is a more significant challenge owing to the limited angle tomography permitted by reflectance-only measurements. We combine previously established concepts for improving image quality and demonstrate, through simulation studies, their application for improving the image quality of adult human brain function. We show in a three-dimensional human head model that localization accuracy is significantly improved by the addition of measurements that provide overlapping samples of brain tissue. However, the reconstructed absorption contrast is significantly underestimated because its depth is underestimated. We show that the absorption contrast amplitude accuracy can be significantly improved by providing a cortical spatial constraint in the image reconstruction to obtain a better depth localization. The cortical constraint makes physiological sense since the brain-activity-induced absorption changes are occurring in the cortex and not in the scalp, skull, and cerebral spinal fluid. This spatial constraint is provided by segmentation of coregistered structural magnetic resonance imaging (MRI). However, the absorption contrast deep within the cortex is reconstructed superficially, resulting in an underestimation of the absorption contrast. The synthesis of techniques described here indicates that multimodality imaging of brain function with diffuse optical imaging and MRI has the potential to provide more quantitative estimates of the total and deoxyhemoglobin response to brain activation, which is currently not provided by either method independently. However, issues of depth resolution within the cortex remain to be resolved. PMID- 15813533 TI - Transferring color between three-dimensional objects. AB - A framework for transferring image-based color between three-dimensional objects by the use of a dichromatic reflection model is proposed. The framework addresses the following issues: (1) accurate recovery of an implicit geometric coefficient, (2) calculation of body color, (3) color transfer between different illuminants, and (4) segmentation of multicolored regions. The experimental results show that high color accuracy and photorealistic effects of the synthesized images can be achieved. The proposed technique has wide applications in image-based design and visualization of three-dimensional objects. PMID- 15813534 TI - Finite-difference time-domain solution of light scattering by an infinite dielectric column immersed in an absorbing medium. AB - The two-dimensional (2-D) finite-difference time-domain (FDTD) method is applied to calculate light scattering and absorption by an arbitrarily shaped infinite column embedded in an absorbing dielectric medium. A uniaxial perfectly matched layer (UPML) absorbing boundary condition is used to truncate the computational domain. The single-scattering properties of the infinite column embedded in the absorbing medium, including scattering phase functions and extinction and absorption efficiencies, are derived by use of an area integration of the internal field. An exact solution for light scattering and absorption by a circular cylinder in an absorbing medium is used to examine the accuracy of the 2 D UPML FDTD code. With use of a cell size of 1/120 incident wavelength in the FDTD calculations, the errors in the extinction and absorption efficiencies and asymmetry factors from the 2-D UPML FDTD are generally smaller than approximately 0.1%. The errors in the scattering phase functions are typically smaller than approximately 4%. With the 2-D UPML FDTD technique, light scattering and absorption by long noncircular columns embedded in absorbing media can be accurately solved. PMID- 15813535 TI - Radiographic human identification using bones of the hand: a validation study. AB - The 1993 Supreme Court case Daubert v. Merrell-Dow Pharmaceuticals, Inc. underscores the importance of validating forensic science techniques. This research examines the validity of using posterior-anterior radiographs of the hand to make positive identifications of unknown human remains. Furthermore, this study was constructed to satisfy the requirements of Daubert's guidelines of scientific validity by establishing a standard methodology for hand radiograph analysis, testing the technique, and noting rates of error. This validation study required twelve participant examiners from the forensic science community, working independently, to attempt to match 10 simulated postmortem radiographs of skeletonized hands to 40 simulated antemortem radiographs of fleshed cadaver hands. The overall accuracy rate of the twelve examiners was 95%, while their collective sensitivity and specificity were 95% and 92%, respectively. However, the accuracy of each examiner was related to the amount of radiological training and experience of the observer. Six Ph.D. forensic anthropologists and four experienced forensic anthropology graduate students correctly identified all the matches. Participant examiners noted bone morphology, trabecular patterns of the proximal and middle phalanges, and distinctive radiopaque and radiolucent features as the anatomical features that aided the identification process. The hand can be an important skeletal element for radiographic positive identification because it contains 27 individual bones for comparative analysis. PMID- 15813536 TI - Prediction of race using characteristics of dental morphology. AB - Historically, forensic anthropology has concentrated on race determination using skeletal morphological variation. Conversely, dental anthropology has been concerned with worldwide patterns of dental morphological variation. This paper represents a synthesis of the goals of forensic anthropology with the methods of dental anthropology. Dentitions of modem African Americans (n = 110) and European Americans (n = 155) were observed using methods based on the Arizona State University dental morphology standards. Of 136 characters observed, eight were found with frequencies that were very different between the groups. Based on these eight characteristics, probability tables were created for determining an individual's social race, using both Bayesian prediction and logistic regression. These tables are applicable for determining the probability that an unknown individual can be categorized as African American or European American. This method was tested on 40 individuals known to belong in one of these two ancestral groups. Correct assignment of race was made in 90% of cases. PMID- 15813537 TI - Symmetrical fracturing of the skull from midline contact gunshot wounds: reconstruction of individual death histories from skeletonized human remains. AB - This paper reports a bilaterally symmetrical cranio-facial fracture pattern that is observed in self-inflicted, midline gunshot wounds. Five cases of self inflicted gunshots wounds are presented as follows: two high-powered rifle cases, two shotgun cases, and one handgun case. In all five cases the remains were either decomposing or skeletonized and submitted to forensic anthropologists. Following identification, the main focus of the anthropological examination was the analysis of perimortem trauma to the skeleton. In each case, the skull was submitted in a highly fragmented state. Nevertheless, by focusing on the pattern of perimortem cranio-facial fractures, the anthropologists contributed key information regarding the circumstances of death. The observed symmetrical cranio facial fracture patterns in the above cases are described in detail and interpreted. The specific location of the linear fractures is discussed, as well as the theoretical rationale behind the location in terms of skeletal architecture, such as buttresses, struts, and sutures. The interpretive framework provided by this paper may prove helpful to others who are faced with similar cases of cranio-facial fracturing. PMID- 15813538 TI - Population variation in femur subtrochanteric shape. AB - Use of proximal femur shape to determine ancestry has appeal, but its validity is problematic because of unaddressed issues associated with skeletal plasticity, within- and between-population variation, sample selection, and interobserver error. In this paper, I inspect within- and between-group variation in proximal femur shape using five groups (American Blacks, American Whites, Hispanics, Native Americans, and Polynesians), and examine the affect of three environmental variables (subsistence strategy, physical terrain, and geographical region). Finally. I consider the validity of using the proximal femur to assess ancestry. The results show that there is significant within-group variation in proximal femur shape. Among Native Americans, both geographical location and subsistence strategy have a significant affect on proximal femur shape. Nevertheless, this study generally verifies the assertion that the proximal femur can be used reliably to distinguish Native Americans from American Blacks and Whites, but its precision may be reduced in some geographical regions. PMID- 15813539 TI - Body height estimation based on dimensions of sacral and coccygeal vertebrae. AB - This study is to evaluate whether it is possible to predict living stature from sacral and coccygeal vertebral dimensions. Individual vertebral body heights, sacral height (SH), and sacrococcygeal height (SCH) were recorded from the magnetic resonance images of 42 adult males. Sum of the heights of five sacral vertebrae (sigmaS), the first four coccygeal vertebrae (sigmaC), and the total height of the sacral and the first four coccygeal vertebrae together (sigmaSC) were also recorded. Linear regression equations for stature estimation were produced using the above mentioned variables. The regression equations were constructed and tested by using jack-knife procedure. Statistical analyses indicated that the combined variables (SH, SCH, sigmaS, sigmaC, sigmaSC) were more accurate predictors of stature than the heights of individual vertebrae. The results of the study pointed out that the equations derived from sacrococcygeal dimensions perform somewhat better than ones based on foot and head variables, but worse than those based on long-bone length. As a conclusion, the dimensions of sacral and coccygeal vertebrae could be used for stature estimation when long bones are not available. PMID- 15813541 TI - A comparison of smokeless powders and mixtures by capillary zone electrophoresis. AB - The analysis of inorganic ions present in smokeless and muzzleloading powders has been performed using capillary zone electrophoresis (CZE). Previous publications have examined inorganic low explosives using CZE, but have not looked at the ion profiles from smokeless powders. In this report, seven commercially available smokeless powders were analyzed as unburned powder and burned residue. The results demonstrate that ionic profiles can be used to characterize smokeless powders. Our analysis also included a smokeless powder/ Pyrodex combination to determine if smokeless powder ions are distinguishable in a mixture; however, the high concentration of ions present in Pyrodex RS prevented its detection. In addition, five different smokeless powder samples as well as Pyrodex RS were collected for analysis subsequent to deflagration in fifteen plastic pipe bombs. The relative ion concentrations between these powders can be used to illustrate the differences between open burning and pipe bomb deflagration. PMID- 15813540 TI - The effect of electron beam irradiation on forensic evidence. 1. Latent print recovery on porous and non-porous surfaces. AB - The recent use of the postal system as a means of delivering anthrax spores via several contaminated envelopes has led to the selective irradiation of mail. These as yet unsolved attacks and the U.S. Postal Service's decision to irradiate certain types of mail has led to some unexpected complications. The high doses of radiation required to destroy biological agents like anthrax are sufficient to induce damage to other materials present in the envelope. There have been reports of damage to many different items that have been subjected to irradiation, including paper, precious gems, plastic, computer discs, and electronics. However, few studies have examined the effect of such treatments on items of forensic interest. In this paper, the authors focused on the impact of the irradiation process on the ability to visualize latent prints. This experiment involved using several donors, substrates (both porous and non-porous), and visualization reagents. The results indicate that the irradiation process can have a detrimental effect on the success of certain visualization reagents. PMID- 15813542 TI - Adsorption saturation and chromatographic distortion effects on passive headspace sampling with activated charcoal in fire debris analysis. AB - Distortion of the chromatographic profile obtained for hydrocarbons that have been sampled by adsorption onto activated charcoal is a well-known phenomenon. The work reported here helps to better define the causes of chromatographic profile distortion and offers a potential method to avoid chromatographic distortion in some cases through a subsampling technique. The recovery of hydrocarbons from an equimolar mixture was investigated to determine the influence of hydrocarbon concentration on the molar ratios of recovered components. In a one-quart container, hydrocarbon volumes as small as 24 microL (liquid) were sufficient to saturate the surface area available for adsorption on a 99.0 mm2 square of activated charcoal, resulting in significant distortions in the molar ratio and the chromatographic profile of the recovered hydrocarbons. Passive headspace sampling of a similarly small volume of unweathered gasoline spiked onto carpet padding resulted in a significant distortion of the chromatographic profile. The chromatographic profile of the recovered hydrocarbons closely resembled 75% weathered gasoline. Heating the container spiked with unweathered gasoline to evenly distribute the components and then removing a subsample of the carpet padding to a second container for passive headspace analysis greatly reduced the amount of distortion in the resulting chromatogram. PMID- 15813543 TI - Simultaneous separation of different types of amphetamine and piperazine designer drugs by capillary electrophoresis with a chiral selector. AB - The recent emergence of a new class of piperazine-type compounds has brought about the need for laboratory screening methods for both seized drugs and toxicological samples. These piperazine compounds, which include 1 benzylpiperazine (BZP) and 1-(3-trifluoromethylphenyl)piperazine (TFMPP), exhibit comparable physiological effects and can be substituted for the classic amphetamine-type drugs. We have optimized a chiral capillary electrophoresis (CE) separation that detects a set of 6 piperazine and 4 chiral amphetamine compounds in under 23 min using a 200 mM phosphate buffer at a pH = 2.8 with 20 mM hydroxypropyl- beta-cyclodextrin (HPbeta3CD). In addition to the above compounds, a series of "clandestine" BZP diHCl samples were also analyzed using this method to assess the ruggedness of the procedure. The novel CE separation was tailored to simultaneously detect these piperzine compounds in addition to amphetamine type drugs. Distinct migration time and UV-spectral data were obtained for all compounds of interest. PMID- 15813544 TI - Liquid chromatography-mass spectrometric and liquid chromatography-tandem mass spectrometric determination of hallucinogenic indoles psilocin and psilocybin in "magic mushroom" samples. AB - Accurate and sensitive analytical methods for psilocin (PC) and psilocybin (PB), tryptamine-type hallucinogens contained in "magic mushrooms," were investigated using liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography tandem mass spectrometry (LC-MS-MS). The chromatographic separation on an ODS column and mass spectral information gave complete discrimination between PC and PB without derivatization. The mass spectrometric detection had a high sensitivity, and the tandem mass spectrometric detection provided more specificity and accuracy, as well as high sensitivity. The detection limits ranged from 1 to 25 pg by LC-MS in the selected ion monitoring mode, and the intra- and inter-day coefficients of variation were estimated to be 4.21-5.93% by LC-MS-MS in the selected reaction monitoring mode. By applying the present LC-MS MS technique to four real samples, the contents of PC and PB were found to vary over a wide range (0.60-1.4 and 0.18-3.8 mg/g dry wt. for PC and PB, respectively) between samples. PMID- 15813545 TI - Forensic significance of bullet lead compositions. AB - The concentrations of seven elements in projectile lead specimens received as evidence were used to assess the frequency of the occurrence of two unrelated samples having indistinguishable compositions. A set of data from 1837 samples was selected for this study from a sampling of 23,054 lead bullets and shot pellets received as evidence in the FBI Laboratory over the period 1989 through 2002. The method used for selection of samples from case submissions ensured that no two samples of the same general type from the same case were included and that no bias was introduced concerning representation of manufacturers or production sources. A total of 1,686,366 pairwise lead sample comparisons were made using the concentrations of the elements Sb, Cu, As, Ag, Bi, Sn, and Cd using a match criterion of two times the sum of the standard deviations of the paired samples. Of the 1837 samples, 1397 samples, or 76%, are distinguishable from every other sample in this study. The total number of indistinguishable sample pairs is 674, for a frequency of 1 out of every 2502 comparisons. The frequency of occurrence of matching samples decreases as the number of measured elements is increased and as the precision of the measurements improves. For bullets in which all seven elements were determined, the match frequency is 1 in 7284. Compositional comparison of bullet lead provides a reliable, highly significant point of evidentiary comparison of potential sources of crime-related bullets. PMID- 15813546 TI - A PCR multiplex and database for forensic DNA identification of dogs. AB - Animal-derived trace evidence is a common finding at crime scenes and may provide an important link between victim(s) and suspect(s). A database of 558 dogs of pure and mixed breeds is described and analyzed with two PCR multiplexes of 17 microsatellites. Summary statistics (number of alleles, expected and observed heterozygosity and power of exclusion) are compared between breeds. Marked population substructure in dog breeds indicates significant inbreeding, and the use of a conservative theta value is recommended in likelihood calculations for determining the significance of a DNA match. Evidence is presented that the informativeness of the canine microsatellites, despite inbreeding, is comparable to the human CODIS loci. Two cases utilizing canine DNA typing, State of Washington v. Kenneth Leuluaialii and George Tuilefano and Crown v. Daniel McGowan, illustrate the potential of canine microsatellite markers for forensic investigations. PMID- 15813547 TI - The importance of thin layer chromatography and UV microspectrophotometry in the analysis of reactive dyes released from wool and cotton fibers. AB - Samples of reactively-dyed wool and cotton were obtained from a range of dye manufacturers, dye distributors and the Forensic Science Service (FSS) Fibre Data Collection. The wool fibers were red in color and had previously been compared using comparison microscopy (CM), visible range microspectrophotometry (VS) and thin layer chromatography (TLC). The cotton fibers were blue and black in color and had not been previously compared. Red, blue and black fibers were chosen because they are often encountered in casework. The usage of reactive dyes to color fibers has increased over the last 10-15 years and these are often seen in casework. Before techniques were available that allowed reactively-dyed fibers to be compared using TLC only CM and microspectrophotometry were routinely carried out. Many laboratories, who had a microspectrophotometer, only had a visible range instrument. It was therefore important to see which techniques provide additional information, that gives greater individuality to fibers, to that obtained from CM. The color was released from the wool and cotton fibres using alkaline hydrolysis and a cellulase enzyme respectively. Many of the red wool samples were differentiated from each other using CM. More differentiation was found using VS and even more when ultraviolet range microspectrophotometry (UV) or TLC was used. Two samples could only be differentiated using TLC because CM, VS and UV failed to separate them. The black cotton samples were predominately differentiated using CM but VS allowed for further differentiation. With the samples used in this project UV and TLC failed to separate the samples further. The blue cotton samples benefited from the use of CM, VS and either UV or TLC to reduce the number of matching pairs. All techniques aided differentiation although with this set TLC and UV proved to be complementary techniques. Results demonstrate that TLC and UV both yield important information over and above that obtained from CM and VS. Although in some parts of the project TLC and UV are complementary if the concentration of the dye in the fiber is not sufficient for TLC or the scientist doesn't wish to 'destroy' the fiber UV would be of more use than TLC. PMID- 15813548 TI - How long after waterproofing a deck can you still isolate an ignitable liquid? AB - Dried, treated wood was sealed with Thompson's WaterSeal "Clear Multi-Surface Waterproofer" and exposed to outdoor, summer conditions. Sections of the sealed wood were then periodically tested and analyzed in accordance with ASTM methods. The WaterSeal contained a medium petroleum distillate (MPD) as a solvent for a wax. The treated lumber contained a background of aldehydes that could mimic a dearomatized MPD if not carefully investigated by the analyst. The MPD of the WaterSeal was detectable 14 days, but not 27 days after application with exposure to relatively hot, dry and sunny weather conditions. The test was repeated with the MPD detectable 16 days, but not 20 days after application with exposure to cooler and wetter weather conditions. The testing demonstrates the need for a thorough and complete investigation by the fire investigator and the submission of comparison samples to the laboratory. PMID- 15813549 TI - Mitochondrial DNA typing screens with control region and coding region SNPs. AB - Mitochondrial DNA (mtDNA) analysis has found an important niche in forensic DNA typing. It is used with highly degraded samples or low-copy number materials such as might be found from shed hair or bones exposed to severe environmental conditions. The primary advantage of mtDNA is that it is present in high copy number within cells and therefore more likely to be recovered from highly degraded specimens. A major disadvantage to traditional forensic mtDNA analysis is that it is time-consuming and labor-intensive to generate and review the 610 nucleotides of sequence information commonly targeted in hypervariable regions I and II (HVI and HVII) of the control region. In addition, common haplotypes exist in HVI/HVII mtDNA sequences that can reduce the ability to differentiate two unrelated samples. In this report we describe the utility of two newly available screening assays for rapid exclusion of non-matching samples. The LINEAR ARRAY mtDNA HVI/HVII Region-Sequencing Typing Kit (Roche Applied Science, Indianapolis, IN) was used to type 666 individuals from U.S. Caucasian, African American, and Hispanic groups. Processing of the LINEAR ARRAY probe panels "mito strips" was automated on a ProfiBlot workstation. Observable variation in 666 individuals is reported and frequencies of the mitotypes within and between populations are presented. Samples exhibiting the most common Caucasian mitotype were subdivided with a multiplexed amplification and detection assay using eleven single nucleotide polymorphisms in the mitochondrial genome. These types of screening assays should enable more rapid evaluation of forensic casework samples such that only samples not excluded would be subjected to further characterization through full HVI/HVII mtDNA sequence analysis. PMID- 15813550 TI - Ear piercing affects earprints: the role of ear piercing in human identification. AB - Previous research conducted into the use of the human ear in the field of forensic identification has focused upon the use of grids and manual methods to measure and catalogue the different anatomical features of the ear. To date, few have considered the importance of the presence of ear piercings and their possible role in human identification. This study aims to highlight the common distribution of piercings of both ears in both genders and to explore the effect of piercings on earprints. The presence of a piercing may, in part, help to explain why partial and not whole earprints are sometimes recovered from a scene of crime (suggesting that the offender's ears may be pierced). The presence of piercings through the tragus and the superior part of the helix are shown to be infrequent and thus may be used to assist the identification of a body, due to its relative rarity with respect to piercings found in other areas of the ear. PMID- 15813551 TI - The applicability of holography in forensic identification: a fusion of the traditional optical technique and digital technique. AB - In this study, the applicability of holography in the 3-dimensional recording of forensic objects such as skulls and mandibulae, and the accuracy of the reconstructed 3-D images, were examined. The virtual holographic image, which records the 3-dimensional data of the original object, is visually observed on the other side of the holographic plate, and reproduces the 3-dimensional shape of the object well. Another type of holographic image, the real image, is focused on a frosted glass screen, and cross-sectional images of the object can be observed. When measuring the distances between anatomical reference points using an image-processing software, the average deviations in the holographic images as compared to the actual objects were less than 0.1 mm. Therefore, holography could be useful as a 3-dimensional recording method of forensic objects. Two superimposition systems using holographic images were examined. In the 2D-3D system, the transparent virtual holographic image of an object is directly superimposed onto the digitized photograph of the same object on the LCD monitor. On the other hand, in the video system, the holographic image captured by the CCD camera is superimposed onto the digitized photographic image using a personal computer. We found that the discrepancy between the outlines of the superimposed holographic and photographic dental images using the video system was smaller than that using the 2D-3D system. Holography seemed to perform comparably to the computer graphic system; however, a fusion with the digital technique would expand the utility of holography in superimposition. PMID- 15813552 TI - Observations on dental structures when placed in contact with acids: experimental studies to aid identification processes. AB - In a "mafia" crime case, a magistrate asked us whether it is possible to destroy a cadaver by immersing it in acids, and would it be possible to identify any residues. The aim of this study was to observe the behavior of teeth exposed to four kinds of acid solutions. The teeth were placed in plastic containers with 25 mL of acid and observed. The experiences showed that teeth are completely dissolved after 14 h of immersion in 37% solution of hydrochloric acid, while at 90h in 96% sulfuric acid, the destruction of the samples is still incomplete. In nitric acid the teeth undergo a complete dissolution in 12 h, and in 17 h in aqua regia (chloroazotic acid-hydrochloric/nitric acid 1:3). It was possible to recognize the characteristic morphological features of dental tissues and structures up until the advanced stages of degradation. PMID- 15813553 TI - Fatal pulmonary thromboembolism and hereditary thrombophilias. AB - Pulmonary thromboembolism (PE) is found commonly in forensic pathology practice, as it typically causes sudden death. It is attributed to a wide variety of predominantly acquired etiologies. Although likely etiologically multifactorial, some common proximate causes include: surgery, pregnancy, injury, inactivity of any cause, cancer, obesity, or serum hyperviscosity. On occasion, no apparent predisposing condition is identified. In these instances, occult hereditary thrombophilias may play a causal role. Deaths referred to the Office of Chief Medical Examiner (OCME) of New York City between December, 2000 and September, 2003 and due to PE were retrospectively reviewed. Molecular analysis (FRET) was performed on selected cases for three common hereditary thrombophilias: mutations in factor V Leiden (FVL), prothrombin G20210A (PT), and methylenetetrahydrofolate reductase (MTHFR). During the study period, 124 of 15,280 deaths were primarily attributable to PE. Of those, 34 were selected for molecular analysis. One or more mutations were detected in 35% of those, five of which were clearly causally related to death. Given the potential benefits to surviving family members, our data indicate that postmortem molecular testing for the common hereditary thrombophilias is warranted in at least selected cases. PMID- 15813554 TI - Unexplained sudden death and the likelihood of drug abuse. AB - The common history of drug abuse in adults with an undetermined cause of death has led us to hypothesize that chronic drug abuse increases the risk of sudden death. To begin evaluating this hypothesis, we conducted a retrospective case control study of 61 decedents whose cause of death remained undetermined following autopsy matched one to one to a control group of pedestrians or passengers killed in motor vehicle collisions. In 21 pairs, the case subject had evidence of drug abuse but the control did not, and in 5 cases the reverse was true. Analysis showed that individuals with an undetermined cause of death are 4.2 times more likely to have evidence of drug abuse than are victims of a motor vehicle collision. PMID- 15813555 TI - Medical malpractice charges in Germany--role of the forensic pathologist in the preliminary criminal proceeding. AB - Medical malpractice charges from 1989 to 2002 were evaluated. A rising number of cases during this period is evident. The charges of practice falling below the standard of care (n = 285) were surveyed to determine who informed the prosecution, which clinical subjects are involved, what kind of charges can be found and whether such allegations can be appropriately assessed by means of a forensic autopsy. Forensic pathologists were found to be useful for ascertainment and interpretation of autopsy findings. If special questions arise, an additional expert opinion should be suggested by the forensic pathologist. There was no relevant shift in the range of subjects involved compared to former studies. The investigated charges might represent only a small fraction of cases of medical practice falling below the standard of care. PMID- 15813556 TI - VIRTOPSY--scientific documentation, reconstruction and animation in forensic: individual and real 3D data based geo-metric approach including optical body/object surface and radiological CT/MRI scanning. AB - Until today, most of the documentation of forensic relevant medical findings is limited to traditional 2D photography, 2D conventional radiographs, sketches and verbal description. There are still some limitations of the classic documentation in forensic science especially if a 3D documentation is necessary. The goal of this paper is to demonstrate new 3D real data based geo-metric technology approaches. This paper present approaches to a 3D geo-metric documentation of injuries on the body surface and internal injuries in the living and deceased cases. Using modern imaging methods such as photogrammetry, optical surface and radiological CT/MRI scanning in combination it could be demonstrated that a real, full 3D data based individual documentation of the body surface and internal structures is possible in a non-invasive and non-destructive manner. Using the data merging/fusing and animation possibilities, it is possible to answer reconstructive questions of the dynamic development of patterned injuries (morphologic imprints) and to evaluate the possibility, that they are matchable or linkable to suspected injury-causing instruments. For the first time, to our knowledge, the method of optical and radiological 3D scanning was used to document the forensic relevant injuries of human body in combination with vehicle damages. By this complementary documentation approach, individual forensic real data based analysis and animation were possible linking body injuries to vehicle deformations or damages. These data allow conclusions to be drawn for automobile accident research, optimization of vehicle safety (pedestrian and passenger) and for further development of crash dummies. Real 3D data based documentation opens a new horizon for scientific reconstruction and animation by bringing added value and a real quality improvement in forensic science. PMID- 15813557 TI - The Richard Cory phenomenon: suicide and wealth in Kansas City, Missouri. AB - This case-control study investigates the relationship between suicide and wealth in Kansas City, Missouri. House and personal property appraisal data on all victims of suicide from 1998 and 2002 and victims from a control population of deaths reported to the Jackson County Medical Examiner during the same time interval were obtained from the Jackson County Government website. The controls were matched to suicide cases by race, gender, year of death, and age at death (+/- 1 year). Data from the 426 members of each group of suicides and controls indicate that suicide victims were: 1) 77% more likely than controls to have lived in houses rather than in apartments or trailers, 2) more likely than controls to have lived in more expensive houses (mean values dollar 70,143 versus dolllar 61,513 respectively, p = 0.04) and 3) more likely to have killed themselves because of factors other than financial strain (8.0% of suicides showed financial strain). PMID- 15813558 TI - Successional behavior and occurrence matrix of carrion-associated arthropods in the urban area of Medellin, Colombia. AB - Forensic entomology is a scientific tool applied to the study of insect or arthropod succession at the scene of a crime or that associated with an accident or natural death. Interpreting this succession provides information to determine minimum and maximum limits of the Postmortem Interval (PMI), that is, the time between death and the discovery of the body. This study was carried out during the rainy season, from 27 October to 12 December 2002 in an urban area of the city of Medellin, Colombia. Three domestic pigs were used as animal models. The results showed that both the aforementioned species as well as the physical characteristics of the carcass determined the indicator species of the postmortem interval in urban areas of the city. In total, 11,937 individuals were collected and identified, belonging to 12 orders, 29 families and 42 genera. Diptera were the most abundant order (90%) represented mainly by Calliphoridae (80%). Coleoptera were scarcely present, representing only 2.8% of the total collected. Based on information obtained of eggs and larvae reared to adult of this successional study, an occurrence matrix was elaborated for determining of the PMI. From the same study area and under the same conditions, a list of arthropods associated with carrion is presented in this manuscript. PMID- 15813559 TI - Postmortem non-invasive virtual autopsy: death by hanging in a car. AB - A body was found behind a car with a noose tied around its neck, the other end of the rope tied to a tree. Apparently the man committed suicide by driving away with the noose tied around his neck and was dragged out of the car through the open hatchback. postmortem multislice-computed tomography (MSCT) and magnetic resonance imaging (MRI) indicated that the cause of death was cerebral hypoxia due to classic strangulation by hanging, and not due to a brainstem lesion because of a hang-man fracture as would be expected in such a dynamic situation. Furthermore, the MRI displayed intramuscular haemorrhage, bleeding into the clavicular insertions of the sternocleidomastoid muscles and subcutaneous neck tissue. We conclude that MSCT and MRI are useful instruments with an increased value compared with 2D radiographs to augment the external findings of bodies when an autopsy is refused. But further postmortem research and comparing validation is needed. PMID- 15813560 TI - Incidental myocardial infarction in Ehlers-Danlos syndrome type IV? AB - Ehlers-Danlos Syndrome Type IV is an illness that often leads to premature death due to arterial rupture or dissection and is characterized by very fragile connective tissue. This report documents the death of a 30-year-old man with Ehlers-Danlos Syndrome Type IV from myocardial rupture and cardiac tamponade following a myocardial infarction. We believe that Ehlers-Danlos Syndrome Type IV contributed to the coronary atherosclerosis and myocardial rupture in this young man and that this disease led indirectly to his death by myocardial infarction, an unusual cause of death in this syndrome. PMID- 15813561 TI - Use of a television crime-drama series to promote legal understanding in mentally ill, incompetent defendants: a pilot study. AB - Videotaped material is used for educational purposes in many areas of medicine. In forensic facilities, programs designed to restore competency to stand trial (CST) in incompetent, mentally ill defendants have utilized videotaped courtroom proceedings as learning tools. This pilot study reviewed the progress of incompetent defendants (N = 15) who participated in a program that utilized videotaped segments of the television crime-drama "Law & Order", among other techniques, to promote CST in individuals deemed unfit to stand trial. The authors hypothesized that participation in at least one cycle of the Competency Restoration Group (CRG)'s curriculum would be associated with improvement in the areas of understanding, reasoning and appreciation. In order to assess whether the group was beneficial to the patient's treatment goal of competency restoration, patients were screened using the MacArthur Competence Assessment Tool-Criminal Adjudication (MacCAT-CA) prior to starting the group and after completing a cycle of the group's curriculum. The Wilcoxon signed ranks test was employed to analyze the results from the pre- and post-group MacCAT-CA testing. The tests yielded significant (p < 0.005) post-test differences in the hypothesized direction for each of the three subsections: Understanding, Reasoning, and Appreciation as well as a significant post test improvement in the total MacCAT-CA scores. These results suggest that a didactic program, using a popular crime drama series, can be effective in facilitating learning in competency restoration programs. Limitations of this study include its lack of a control group and small population. PMID- 15813562 TI - Drug and alcohol use as determinants of New York City homicide trends from 1990 to 1998. AB - In this population-level study, we analyzed how well changes in drug and alcohol use among homicide victims explained declining homicide rates in New York City between 1990 and 1998. Victim demographics, cause of death, and toxicology were obtained for all homicide (N = 12573) and accidental death victims (N = 6351) between 1990 and 1998 from the Office of the Chief Medical Examiner of New York (OCME). The proportion of homicide and accident decedents positive for cocaine fell between 1990 and 1998 (13% and 9% respectively); the proportion of homicide and accident decedents positive for opiates and/or alcohol did not change significantly. Changing patterns of drug and alcohol use by homicide victims were comparable to changing patterns of drug and alcohol use in accident victims, suggesting that changes in drug and alcohol use among homicide victims between 1990 and 1998 cannot solely explain the decline in NYC homicide rates. PMID- 15813563 TI - Population genetics of two STR loci D13S796 and DYS439 in a Chinese Han population. PMID- 15813564 TI - Allele frequencies for three STR loci D2S2952, D3S1744, D12S1294 in Chinese population. PMID- 15813565 TI - Genetic data of five STR loci in a population sample of Santa Catarina, Brazil. PMID- 15813566 TI - Distribution of D7S821, D15S642, and GATA184A08 alleles in a Chinese population sample. PMID- 15813567 TI - Polymorphism study on nine STRs loci of Han ethnic group in Hanzhong region (West of China). PMID- 15813568 TI - Data on eight STR loci in Shan, Akha, Lisu, Lahu, and Hmong populations of Northern Thailand. PMID- 15813569 TI - How to achieve "WOW" customer service. PMID- 15813570 TI - A review of dental suturing for optimal soft-tissue management. AB - Establishing nontension primary wound closure of various soft-tissue flaps is paramount for optimal postsurgical wound healing. Surgical procedures that require clinical flap manipulation such as those used with traditional periodontal therapy, periodontal plastic cosmetic surgery, hard- and soft-tissue regeneration, and the excision of pathologic tissue also require excellence in execution and thorough understanding of the various techniques of surgery, suturing, and the materials currently available to ensure the desired clinical results. This article will discuss the rationale of specific suturing techniques and suture materials to aid the clinician in obtaining optimal wound closure. PMID- 15813571 TI - Maximizing esthetic transformations using a closed flap ErCr:YSGG modality. PMID- 15813572 TI - The use of anxiolytic medications to supplement local anesthesia in the anxious patient. AB - Local anesthetic failures in dental patients can have many causes, including anatomical variations, technique, and anxiety/fear. By understanding the mechanisms responsible for failed local anesthesia, patients can be treated more comfortably. The interaction of anxiety and fear is discussed. Oral sedation dentistry is highlighted as a way to reduce anxiety/fear and the patient's perception of pain. Profound anesthesia can be accomplished more easily in relaxed patients with diminished or eliminated anxiety/fear. PMID- 15813573 TI - Updated CDC infection control guidelines for dental health care settings: 1 year later. PMID- 15813574 TI - Universal paradigms for predictable final impressions. AB - The master blueprint for indirect restorations is the final impression. The challenge for the clinician is to establish a protocol that ensures a predictably excellent final impression. The purpose of this article is to provide a protocol that integrates the many detailed steps of impression making, from patient comfort to dental laboratory communication. Understanding the biology of the dentogingival junction, dental materials and their interactions, and proper technique all contribute to the final results. PMID- 15813575 TI - New technologies: quackery or fraud? AB - A few dentists are responsible for most of the unconventional or unethical dentistry performed on the public. However, the influence of a few can have a major effect on the public's trust of the profession. Suppressing the introduction or continued development of new technologies in the dental profession is not the solution. Rather, new technologies should be integrated into existing established modes of dental practice. Innovations are good for the dental profession and for the public. As prudent dentists, our goal should be to critically assess innovations and demand validation before allowing unconditional widespread use in dental procedures. Regarding unethical clinicians, they probably always will put their interest ahead of the patient's. However, through peer pressure and enhanced education in ethics early in dental education, we can begin to minimize such activity within the profession. PMID- 15813576 TI - Endodontic synchronicity. AB - This article examines a new rotary technique in endodontic instrumentation and stresses the importance of adhering to basic principles. Straight line access and its clinical implication to endodontic success is reviewed. A new system of nickel titanium rotary instruments that uses a reamer design (EndoSequence) is introduced and its associated design features are thoroughly discussed. A step-by step description of the clinical application of this new methodology is presented. PMID- 15813580 TI - Clarification of social support. AB - PURPOSE: To clarify the concept of social support. DESIGN: Template Verification and Expansion Model. METHODS: Meta-synthesis strategies. FINDINGS: Social support is composed of emotional and instrumental support. It is an advocative interpersonal process characterized by reciprocal exchange of information, it is context specific, and it results in improved mental health. Antecedents of emotional and instrumental support include a perceived need plus a social network and climate that are conducive to the exchange of social support. CONCLUSIONS: In keeping with the nonprofessional nature of social support, nurses are advised to encourage patients to use and enhance personal support networks. For the same reason, nurses should reconsider investigating social support as a nursing intervention. Expanded efforts are needed to differentiate social support from concepts such as caring. PMID- 15813581 TI - Action responses of Congolese refugee women. AB - PURPOSE: To describe Congolese refugee women's action responses to difficult living situations. DESIGN: Interpretive qualitative. METHODS: Narrative data were collected from 14 Congolese refugee women using minimally structured interviews and were analyzed for patterns in their responses to their difficult situations. FINDINGS: The narratives about participants' difficult circumstances indicated six action responses: refiguration, advocacy, resistance, resignation, sorrow, and faith. CONCLUSIONS: When encountering difficult life circumstances, refugee participants demonstrated actions with varying influence on their agency. PMID- 15813582 TI - Disclosing genetic test results to family members. AB - PURPOSE: To describe the experiences of disclosing genetic test results to biological family members among people tested for Huntington's disease (HD) or hereditary breast and ovarian cancer (HBOC). DESIGN: Grounded theory methodology. METHODS: Open-ended, tape-recorded interviews were conducted with 29 participants 24 who had received genetic test results and 5 who had decided not to be tested. The participants were from three countries, including 15 U.S. states. Interviews occurred from 2 months to 4 years after receiving test results. Tapes were transcribed and analyzed for conceptual categories to describe the experience of disclosing genetic test results. FINDINGS: Participants described the effects and meaning of disclosing test results to their various family members, and they selectively disclosed results to family members. The timing of disclosure was influenced by the particular disease and the person's perceived need to prepare. Disclosure of genetic test results brought the risk of HD and HBOC to the foreground, not only for the person tested but for family members as well. CONCLUSIONS: This study elucidated the perspective of the "discloser" and the consequences they anticipated and experienced. PMID- 15813583 TI - Nonadherence with diet and fluid restrictions among adults having hemodialysis. AB - PURPOSE: To describe the prevalence of nonadherence with the prescribed diet and fluid restrictions among patients receiving hemodialysis. METHODS: A multicenter cross-sectional design with 916 patients recruited from 18 dialysis centers in Germany and Belgium. The Dialysis Diet and Fluid Nonadherence Questionnaire (DDFQ) was used to measure patients' nonadherence. FINDINGS: The results showed that many patients had difficulty following diet (81.4%) and fluid (74.6%) restrictions. Younger male patients and smokers were at highest risk for non adherence. Higher levels of interdialysis weight gain were associated with nonadherence. CONCLUSIONS: The findings indicate the need to continue to monitor and study hemodialysis patients' adherence behavior longitudinally and to design interventions to enhance adherence. PMID- 15813584 TI - Nurse counseling of patients with an overconsumption of alcohol. AB - PURPOSE: To identify the associations between self-reported alcohol-related clinical practice among hospital nurses and their (a) self-rated qualifications for counseling on alcohol, and (b) attitudes toward care of patients with an overconsumption of alcohol. DESIGN AND METHODS: A survey based on self administered questionnaires was undertaken at a Danish University Hospital in the Copenhagen area. Descriptive statistics, chi-square test, and multiple logistic regression analyses were used. Variables were self-reported alcohol-related counseling, self-rated qualifications for counseling patients with an overconsumption of alcohol, attitudes toward the priority of counseling on alcohol and other lifestyle issues, and attitudes toward caring for patients with an overconsumption of alcohol. FINDINGS: Self-rated qualifications for counseling and attitudes toward care significantly influenced the self-reported clinical care of patients with an overconsumption of alcohol. Perceived self-efficacy was positively associated with frequent counseling and positive attitudes toward prevention. Psychiatric nurses were most active, medical nurses less active, and surgical nurses least active in counseling. CONCLUSIONS: To promote nurse involvement in alcohol prevention, increasing the capacity for intervention via skill-based training on assessment of alcohol problems and intervention techniques is necessary. All nurses, but especially surgical nurses, needed updates on alcohol-related counseling. PMID- 15813585 TI - Translation and validation of the condom self-efficacy scale with Thai adolescents and young adults. AB - PURPOSE: To translate the 14-item Condom Self-Efficacy Scale (CSES) into Thai and to validate the Thai version of the Condom Self-efficacy Scale (CSES-T) among Thai adolescents and young adults. METHODS: The CSES was translated using a back translation technique and validated with a cluster-based sample of 425 participants aged 18 to 22 years from eight randomly selected private vocational schools in Bangkok. Participants completed anonymous self-administered scales. Principal component analysis with varimax rotation was conducted to identify latent factors. RESULTS: Factor analysis indicated three factors: communication, correct use, and consistent use. Items loading on the original CSES also loaded on the same factors of the CSES-T except one item. The Cronbach's alpha coefficients were .85 for the total scale, .70 for consistent use, .79 for correct use, and .80 for communication. CONCLUSIONS: Based on psychometric properties, the CSES-T is a valid and reliable tool. It is culturally appropriate for Thai young adults. Thai researchers and health care providers can use the CSES-T to assess adolescents' and young adults' self-efficacy to use condoms as well as to further develop and evaluate interventions to increase condom use. PMID- 15813586 TI - Reproductive decision-making among HIV-positive couples in Taiwan. AB - PURPOSE: To explore the gender-based power relationships and social and cultural influences on reproductive decision-making processes among HIV-positive couples in Taiwan. METHODS: The study sample consisted of 14 HIV-positive couples in southern Taiwan. Data were analyzed using feminist ethnography to explore reproductive decisions made by these couples within the context of Taiwanese society. FINDINGS: Self-knowledge of HIV status had a limited influence on decisions about childbearing. More important was the Confucian values of filial piety and familial obligation, a powerful norm that still dictates procreation decisions in Taiwan. The process by which the couples made reproductive decisions consisted of four stages: initial reproductive decisions between partners, their search for information, their encounters with medical systems, and their weighing risks and benefits. Male and female partners expressed different concerns, and gender-based power relationships were exercised during the decision-making process. CONCLUSIONS: HIV status was not the sole determinant of reproductive decisions made by HIV-positive couples. Rather, the Confucian value of filial piety drove the couples' reproductive decisions. PMID- 15813587 TI - Mothers' experience of helping young adults with traumatic brain injury. AB - PURPOSE: To describe mothers' experience of helping young adults with traumatic brain injury (TBI). DESIGN: Descriptive. METHODS: A convenience sample of participants from support groups for parents of young adults with TBI met the criteria of engaging in regular interaction or helping their children (aged 20 to 36 years). These young adults had suffered moderate or severe TBI from a motor vehicle collisions, sports-related injuries, or recreation-related injuries more than 6 months earlier. A descriptive phenomenological method was used. Three in depth interviews were done with each mother over a 2-month period. Data were the mothers' perceptions, actions, and intentions pertaining to their experiences of helping the young adults. FINDINGS: Five phenomena that were structures of the experience were discerned, discussed with participants to obtain their feedback, and compared to the relevant literature. The five phenomena of the mothers' experiences were: reconnecting my child's brain, considering my child's safety, making our lives as normal as possible, dealing with our biggest problem, and advocating for my child. CONCLUSIONS: The mothers continued rehabilitation efforts with the young adults, even when only minimal services were available to support their efforts. Mothers needed interventions to enhance their knowledge, and they and the young adults with TBI needed expanded community services. PMID- 15813588 TI - Describing use of nursing interventions for three groups of patients. AB - PURPOSE: To describe the nursing interventions used most frequently during an acute hospital stay for three patient groups. METHODS: Analysis of nursing interventions documented by use of the Nursing Interventions Classification (NIC) contained in an electronic clinical database obtained from 33 general inpatient units in one large health care facility from July 1, 1998, to June 30, 2002. The numbers of hospitalizations for each sample in the analyses were: 1,435 in the heart failure group, 567 in the hip fracture procedures group, and 11,756 in the fall prevention group. FINDINGS: The mean number of interventions done at least once during a single hospitalization ranged from 18 to 22 for the three samples. For the total number of patient hospitalizations for each group, the number of interventions ranged from 94 for the hip fracture procedures sample to 182 for the fall prevention sample. Seven interventions were done twice or more a day in at least 20% of the sample in all three groups. Patterns of interventions during the first 6 days of care differed by intervention and sample, indicating that nursing care was individualized. CONCLUSIONS: The results indicate the types of information that can result from analysis of actual clinical nursing data documented with standardized language (Nursing Interventions Classification) in a nursing information system. The knowledge of nursing interventions used in clinical practice has major implications for staff development and nursing education. The information is also useful in making staffing decisions for different types of patient populations. PMID- 15813589 TI - Organizational attributes valued by hospital, home care, and district nurses in the United States and New Zealand. AB - PURPOSE: To determine whether hospital-based, home care, and district nurses identify a core set of organizational attributes in the nursing work environment that they value as important to the support of professional practice. DESIGN: Survey data, collected in 2002-2003 from 403 home care nurses in the United States (US) and 320 district nurses in New Zealand (NZ), were pooled with an existing data set of 669 hospital-based nurses to conduct this descriptive, nonexperimental study. METHODS: The importance of organizational attributes in the nursing work environment was measured using the Nursing Work Index-Revised (NWI-R). Frequency distributions and analysis of variance were used to analyze the data. FINDINGS: At least 80% of hospital-based, home care, and district nurses either agreed or strongly agreed that 47 of the 49 items comprising the NWI-R represented organizational attributes they considered important to the support of their professional nursing practice. Mean importance scores among home care nurses, however, were significantly lower than were those of the other two groups. CONCLUSIONS: Overall, hospital-based, home care, and district nurses had a high level of agreement regarding the importance of organizational traits to the support of their professional practice. The intensity of the attributes' importance was less among home care nurses. Further research is needed to determine whether this set of organizational traits, measured using the NWI-R, is associated with positive nurse and patient outcomes in home care and district nursing practice, as has been shown in acute care settings. PMID- 15813590 TI - Hospital readmission from home health care before and after prospective payment. AB - PURPOSE: To describe and compare clients who were readmitted to the hospital during an episode of home health care, before and after the inception of the prospective payment system (PPS). DESIGN: A longitudinal mixed design was used to replicate a study conducted 9 years previously (pre-PPS) in the same home care agency in the central part of the United States. METHODS: Seventy-six closed-case medical records from a not-for-profit hospital-affiliated home care agency were retrospectively reviewed and compared to pre-PPS data. The same data collection tool, the Hospital Readmission Inventory, was used for both pre- and post-PPS studies. Nurse administrators at the data collection agency were interviewed concerning comparative results. FINDINGS: Currently readmitted clients were sicker than were those in the previous research report, they were readmitted sooner for a different diagnosis, and they had less continuity of services. CONCLUSIONS: The home health care industry has undergone a dramatic change in payment for services, from fee-for-service to PPS. Of particular concern is the adverse patient outcome of an unplanned hospital readmission. Prior studies have characterized such patients in home health care, but no comparative reports were found in a literature search since the inception of PPS. Findings from this study indicated that an increased emphasis on cost containment and higher-risk clients appear to have changed patterns of care delivery. PMID- 15813591 TI - Emergency department and community health center visits and costs in an uninsured population. AB - PURPOSE: To assess how a Community Health Center (CHC) influenced access to care for uninsured people and to describe the economic effect on the local hospital. DESIGN: A framework on access to health care was used in this community-level, descriptive study. METHODS: Data were collected on emergency department (ED) use before state funding of the CHC, 1988 and through 2001. Information included insurance status, charges, diagnosis, and complexity of services received. This study is a population study, and descriptive statistics were used to analyze the data. FINDINGS: Within 3 years after state funding of a CHC began, uninsured visits to the local hospital ED decreased by almost 40%. After 10 years, uninsured ED visits remained 25% lower than before state funding began, whereas insured visits had almost doubled (98%). The decrease in number of uninsured visits saved the hospital and uninsured patients almost $14 million. CONCLUSIONS: After establishment of the CHC, ED visits by uninsured patients declined. Although a causal link cannot be made between the CHC and ED, the descriptive data provide information about a linkage between the decline of uninsured ED visits and the CHC. PMID- 15813592 TI - An international survey of nurse editors' roles and practices. AB - PURPOSE: To describe the editorial practices of nurse editors, including proprietary arrangements, manuscript processing systems, and editorial review, and to ascertain editors' opinions on effective editorial practices. DESIGN: A descriptive study in which an author-designed 108-question survey was distributed and collected by e-mail. Of 177 international nursing editors identified, 164 e mail addresses could be located, and 137 nurse editors expressed willingness to participate. Ninety journals were represented in the final purposive convenience sample, including 71 published in the US and 19 outside the US. Data were analyzed using descriptive statistics, t test, chi square, and content analysis. FINDINGS: Of the 90 journals, 78 were characterized by their editors as scholarly journals. Fifty-two were official journals of associations. The nurse editors' average age was 53, with over 20 years of experience in nursing and 8 years as editor. Although practices and compensation at U.S. and international journals did not differ significantly, scholarly journals and "other" publications such as magazines and newsletters were managed differently. Blinded peer review was in place at 98% of journals. Editors generally used manuscript reviewers as advisors rather than as voters; only 30% of editors thought their ultimate decision must agree with the majority of reviewers. Nurse editors often needed 3-5 years to feel comfortable in the role. Editors of association journals often reported struggling with the issue of editorial independence versus association control. Editors believed their role was influential in maintaining scholarly excellence and evidence-based practice, but many noted the constant pressure of deadlines and dealing with poor writing from authors as challenges of the work. CONCLUSIONS: The role of nurse editor is not part of the formal preparation of nurse scholars or clinicians; standards for nurse editors are not readily apparent, and nurse editors often work in isolation, lacking professional support. This survey included data by which nurse authors and readers of nursing journals can understand how nurse editors work and make decisions about manuscripts, ultimately influencing knowledge dissemination in nursing journals. Nurse editors can use this information to compare their practices with that of others, and advocate for adequate preparation, compensation, and assistance with production. PMID- 15813593 TI - Psychometric approaches to the identification of LD: IQ and achievement scores are not sufficient. AB - Simulated data were used to demonstrate that groups formed by imposing cut-points based on either discrepancy or low-achievement definitions of learning disabilities (LD) are unstable over time. Similar problems were demonstrated in longitudinal data from the Connecticut Longitudinal Study, where 39% of the children designated as having LD in Grade 3 changed group placement with repeated testing in Grade 5. These results show that the practice of subdividing a normal distribution with arbitrary cut-points leads to instability in group membership. Approaches to the identification of children as having LD based solely on individual test scores not linked to specific behavioral criteria lead to invalid decisions about individual children. Low-achievement definitions are not a viable alternative to IQ-discrepancy definitions in the absence of other criteria, such as the traditional exclusions and response to quality intervention. If we accept the premise of multiple classes of low achievers, then we must develop identification systems that are valid and abandon systems whose only merits are their historical precedence and convenience. PMID- 15813594 TI - Male vulnerability to reading disability is not likely to be a myth: a call for new data. AB - Whether boys are more vulnerable than girls to reading disabilities (RD) is controversial. We review studies that were designed to minimize ascertainment bias in the selection of individuals with RD. These include population-based studies that identified children with RD by objective, unbiased methods and studies that examined the gender ratios among the affected relatives of those diagnosed with RD. We conclude that even when ascertainment biases are minimized, there is still a significant preponderance of boys with RD, although the gender ratio of the affected relatives of those with RD manifests the weakest male bias. Furthermore, we demonstrate that potentially confounding factors such as attentional or neurological problems, race, IQ, and severity of RD cannot account for the observed gender bias. We end with a clarion call to future researchers to (a) consider analyzing gender differences by means of more than one definition of RD, (b) compare gender ratios when boys and girls are ranked against the performance of their own gender as opposed to an average across genders, and (c) report group differences in variability and effect sizes of obtained gender ratios. PMID- 15813595 TI - Learning disabilities in children with very low birthweight: prevalence, neuropsychological correlates, and educational interventions. AB - This study examined achievement, neuropsychological, and intervention outcomes at a mean age of 11 years in children with very low birthweight (VLBW, <1500 g) compared with a term-born control group. To assess the prevalence and correlates of specific learning disabilities (LD), the sample was limited to children without neurosensory disorders who had at least low average IQ. Participants included 31 children with <750 g birthweight, 41 with 750-1499 g birthweight, and 52 controls. The <750 g group obtained lower scores in math, IQ, and perceptual organizational skills than the term-born group. The < 750 g group also had higher rates of past and present LD than the controls. Despite these differences, the groups did not differ significantly in rates of special education or tutorial/remedial assistance. These findings suggest that children with extremely low birthweight without gross physical or intellectual impairments are at higher risk for LD and cognitive deficiencies than their term-born peers and that further efforts are needed to improve identification and treatment of these learning problems. PMID- 15813596 TI - The diversity of attention deficits in ADHD: the prevalence of four cognitive factors in ADHD versus controls. AB - The performance of participants with attention-deficit/hyperactivity disorder (ADHD) relative to control participants was measured on four tasks uniquely assessing the functions of selective attention, executive attention, sustained attention, and orienting of attention. The results showed that deficits in sustained attention were the most pronounced, characterizing most participants with ADHD and deficits in each of the other three functions characterized more than half of these participants. Different participants with ADHD revealed different clusters of attentional deficits. These results call for a revision of leading theories of ADHD that identify the core of the pathology as a sole deficit in executive functions. PMID- 15813597 TI - Studying disability and disability studies: shifting paradigms of LDA synthesis of responses to Reid and Valle. AB - In this article, I discuss the 11 diverse responses to Reid and Valle's work on the discursive practice of learning disabilities (LD), implications for instruction, and parent-school relations. I highlight key ideas from each article and then focus on three common areas of interest shared by most respondents: the unacceptable status quo of schooling practices; desired changes in schooling practices; and knowledge production in the field of special education and its relation to schooling practices. In light of the many issues raised, I urge the need for a reflective turn in the field of LD and for openness toward diversification of thought. PMID- 15813598 TI - Discourses on bad children and bad schools. AB - First, discourse is discussed as concerning the subtler lineaments of knowledge production in notions about difference and difficulty. The unhelpful influences that these notions have had on the development of special education research and practice are discussed. The importance of the scholar-practitioner in undoing some of these understandings is further developed. Second, the point is stressed that politics and political questions are intertwined with empirical questions and that a discourse of objectivity propagates and fosters a belief that the supposed paraphernalia of science (measurement, experiment, prediction) can be employed without cost. Third, the point is made that the individual discourse is rooted in the theoretical context of special education--with unhelpful consequences for the way that difficulty continues to be construed as rooted in individuals, whether those individuals be children or schools. PMID- 15813599 TI - Learning disabilities and the new reductionism: a response to Reid and Valle. AB - The purpose of this article is to respond to Reid and Valle's article "The Discursive Practice of LD: Implications for Instruction and Parent-School Relations" in this issue. Our response to the discursive analysis is organized around two major themes: (a) the issue of balance in the scholarship on learning disabilities (LD) and (b) the centrality of the discourse of general education to the discourse of LD. Following our response to the discursive analysis, we comment on the implications for instruction and parent-school relations drawn by Reid and Valle. Highlighted in this section is the disconnect between the macro level findings from the discursive analysis and the micro-level orientation of their "sociopolitical vision" for instruction and parent-school relations. PMID- 15813600 TI - Childhood precursors of adolescent outcomes in type 1 diabetes mellitus. AB - Contemporary outcome measures of chronic illnesses such as type 1 diabetes mellitus are broader than those clinical outcomes traditionally assessed in therapeutic encounters. A holistic approach emphasises quality of life and emotional well-being as well as the achievement of optimal disease management. This paper reviews current knowledge about growth, metabolic control, diabetes complications, neurocognitive and psychological outcomes as well as health related quality of life in childhood diabetes mellitus. It is suggested that the antecedents of adverse diabetes and psychological outcomes in adolescence lie in the years prior to adolescence. The model of care in childhood diabetes mellitus must be focussed on earlier screening and intervention if adverse outcomes are to be reduced. PMID- 15813601 TI - Normal male childhood and adolescent sexual interactions: implications for sexual orientation of the individual with intersex. AB - Data provided by 24 adult men, 20 heterosexual and four homosexual, concerning parental, religious, geographic and explicit sexual innuendos, comments and childhood experiences are presented and discussed in an attempt to consider some of the multiple factors impacting the development of sexual orientation. All of the study subjects were normally developed males and were presumed to have been exposed to normal male levels of androgens prenatally. Since the experiences and perceptions reported are conditioned by a unique social environment that has been superimposed on a normal male typical prenatal CNS differentiation, the experiences of these men suggest that affirmation of masculinity, and openness in the realm of social and sexual interaction, may enhance the formation of a heterosexual orientation. Conversely, sexually explicit feedback with critical implications occurred commonly among the homosexual men, which they interpreted as implying an insufficient masculinity. Both innate factors and social influences impact sexual orientation; in some instances males appear to have been homosexual from early childhood onward, while in other cases there appears to have been some degree of conditioning and choice in sexual orientation. Regarding the intersexed male, this suggests that social interactions, particularly those provided by parents, have a major influence on the development of sexual orientation in the child, while all persons involved in these children's lives and particularly those who nurture must be prepared for any sexual orientation that develops. PMID- 15813602 TI - 5alpha-reductase 2 gene mutations in three unrelated patients of Greek Cypriot origin: identification of an ancestral founder effect. AB - INTRODUCTION: 5alpha-Steroid reductase deficiency (5alphaSRD) is an autosomal recessive enzymatic deficiency. Mutations in the 5alpha-steroid reductase type 2 gene (SRD5A2) result in male pseudohermaphroditism caused by decreased dihydrotestosterone (DHT) synthesis--a key hormone of virilization of male external genitalia. AIM: To study for the first time patients from the Greek Cypriot population, describe their clinical characteristics, and identify the genetic mutations of the SRD5A2 gene. PATIENTS AND METHODS: Three unrelated patients with 46,XY karyotype born with ambiguous genitalia were examined. Patient 1 was raised as a girl and was diagnosed with partial androgen insensitivity syndrome, based on the clinical picture and incomplete laboratory investigation at the age of 4 years, and underwent gonadectomy. For this patient sequencing analysis of all five exons of the SRD5A2 gene and exons 2 to 8 of the androgen receptor (AR) gene was performed. Patients 2 and 3 were also born with ambiguous genitalia. The hCG test for these two patients was informative of 5alphaSRD, as it showed elevated T/DHT ratio after stimulation. Despite genetic counseling, both families decided to raise their infants as females because of severe under-virilization. Sequencing of the SRD5A2 gene was also completed for both patients. RESULTS: No mutations were found in the AR sequence for patient 1. Patients 1 and 3 were found homozygous for the mutation A/G at splice junction intron 1/exon 2 and patient 2 was found heterozygous for the same A/G substitution and also heterozygous for an additional mutation, Pro181Leu, in exon 3. CONCLUSIONS: The same mutation in the SRD5A2 gene was identified in three unrelated patients, in both homozygous and heterozygous form. This splice mutation was previously reported in Turkish patients. This underlying genetic abnormality may be characteristic for the Eastern Mediterranean region and is likely due to an ancestor effect. PMID- 15813603 TI - Rates of diabetes mellitus-related complications in a contemporary adolescent cohort. AB - BACKGROUND: Ten years after the Diabetes Control and Complications Trial there is a paucity of data as to what are current rates of diabetes-related complications in adolescence. OBJECTIVE: To assess the incidence of diabetes-related complications in a contemporary cohort of adolescents with type 1 diabetes mellitus. DESIGN: Retrospective cross-sectional survey. PATIENTS: Adolescents aged >10 years with type 1 diabetes mellitus for >5 years from the Royal Children's Hospital, Melbourne Diabetes Clinic. RESULTS: 382 patients were studied (191 male). The mean HbA1c for males was 8.72% and for females was 8.80%. The rates of hypothyroidism and hypercholesterolaemia were 1.5% and 22% respectively. Twenty-five patients (8%) had intermittent microalbuminuria and six (2%) had persistent microalbuminuria. Only one patient had macroalbuminuria (0.3%). Only two patients (0.7%) with mild non-were diagnosed proliferative diabetic retinopathy. Coeliac disease was diagnosed in 6% of patients. CONCLUSIONS: In this representative and contemporary cohort of diabetic adolescents the incidence of microvascular diabetes-related complications is quite low. PMID- 15813604 TI - Lipoprotein composition in children and adolescents with type 1 diabetes mellitus. AB - Atherosclerotic cardiovascular diseases are the major causes of morbidity and mortality in patients with diabetes mellitus. Both quantitative and qualitative abnormalities of lipo-proteins are associated with the development of atherogenesis. In this study, the prevalence of dyslipidemia and the relative levels of glycosylated lipoproteins in 20 children and adolescents with type 1 diabetes mellitus were determined. Lipid profile, apolipoproteins A-I and B, Lp(a) and LpA-I in plasma were assayed. LpB and glycosylated HDL and LDL were evaluated by ELISA. Diabetic patients and controls had normal lipid profiles, but the diabetic group showed significantly higher LpA-I and lower LpA-I:A-II concentrations than controls. The diabetic group showed a significantly higher glycosylation level of HDL than controls and did not show a statistical difference for glycosylated LDL. No significant correlation between glycosylated lipoproteins, glycemia or HbA1c was found. In conclusion, these results suggest that type 1 diabetic patients develop important qualitative lipid abnormalities. PMID- 15813605 TI - Knowing more by fewer measurements: about the (In)ability of bioelectric impedance to enhance obesity research in children. AB - The prevalence of obesity is increasing worldwide. The implications for human health can already be observed in children. Consequently, it is desirable to provide good quantitative descriptions of the relationship of body fat and health risks, such as hypertension. Bioelectric impedance analysis has been frequently praised to be useful for assessing body fat. Devices to analyse body composition based on this technique seem to be selling well, while the real gain in information they provide is unclear. Here we show in a cohort of 2,218 schoolchildren that the body mass index (which is more easily and less costly to determine) is a better predictor of hypertension than the data delivered by impedance analysis. Moreover, we demonstrate that the output of a random number generator is competitive with impedance measurement for this purpose. It is explained by simple arguments from physics why the formulas for the computation of body fat from bioelectric impedance obliterate rather than clarify the relationship of obesity and hypertension. As a consequence, we suggest questioning the opinion propagated by others that bioelectric impedance analysis is a useful tool in field studies on body fat in children. Measurements requiring more effort (compared to simpler methods) should be proved to add worthwhile information, otherwise they should be avoided. PMID- 15813606 TI - Relationship between plasma leptin, insulin and tumor necrosis factor alpha in obese children. AB - OBJECTIVES: 1. To evaluate the relationship between plasma leptin and TNFalpha concentrations in obese children and to assess the differences between hyperinsulinemic and normoinsulinemic groups. 2. To evaluate the relationship between plasma leptin and insulin levels in obese children. 3. To investigate the TNFalpha G308A mutation in obese children. METHODS: Body mass index (BMI), fasting plasma glucose and insulin levels, oral glucose tolerance test results, homeostasis model assessment of insulin resistance (HOMA-IR) results, and plasma leptin and TNFalpha concentrations were evaluated in obese children (n = 45) and age- and gender-matched, lean healthy controls (n = 40). RESULTS: In obese children the fasting insulin, HOMA-IR results, plasma leptin and TNFalpha concentrations were significantly higher than in controls (p <0.05). Furthermore, obese females showed higher plasma leptin and insulin resistance compared to obese males. While plasma leptin, TNFalpha levels and HOMA-IR results were similar in the prepubertal and pubertal groups, insulin levels were significantly higher in the pubertal group. Plasma leptin and TNFalpha concentrations were similar in hyperinsulinemic and normoinsulinemic obese children. In control children, plasma leptin concentrations showed a positive correlation with BMI, age, fasting insulin and HOMA-IR results. In obese children, plasma leptin levels did not correlate with BMI, fasting insulin or TNFalpha. CONCLUSION: Plasma leptin concentrations did not show any correlation with TNFalpha levels in obese children. Furthermore, plasma leptin and TNFalpha concentrations were similar in hyperinsulinemic and normoinsulinemic obese children. PMID- 15813607 TI - The effect of letrozole on bone age progression, predicted adult height, and adrenal gland function. AB - A common problem in pediatric endocrinology is limited growth potential resulting from advancing skeletal maturation. We determined the efficacy of letrozole, an aromatase inhibitor, on delaying bone age advancement in adolescent males with limited growth potential. Twenty-four patients met the study inclusion criteria. Six patients treated with androgen were analyzed separately. Low-dose ACTH stimulation tests were performed to ascertain the effect of letrozole on adrenal gland function. In patients not on androgen, bone age progression decelerated from 1.51+/-0.57 (deltabone age/deltachronological age) before treatment to 0.68+/-0.66 on therapy (mean duration 12.4 months; p <0.0005). Predicted adult height standard deviation scores (SDS) increased from -1.41+/-0.54 to -0.64+/ 0.65 on treatment (p <0.0005). Similar results were noted in androgen-treated patients. Approximately one-fourth of patients displayed subnormal responsiveness to ACTH. In summary: 1) letrozole decelerates skeletal maturation, resulting in significant increases in predicted adult height, and 2) letrozole causes mild adrenal suppression. PMID- 15813608 TI - Disturbed adrenal function in adolescents with chronic fatigue syndrome. AB - OBJECTIVE: To investigate adrenal function in children and adolescents with chronic fatigue syndrome (CFS) compared with age-matched controls. METHODS: Case control study of low dose (500 ng/m2) synacthen tests (LDST) in 23 adolescents with CFS and 17 age-matched controls. Serum cortisol concentrations were measured at 5-min intervals from 10 to 45 minutes. Peak serum cortisol concentration, time to peak, rise in cortisol and area under the curve (AUC) were derived. RESULTS: Patients with CFS had significantly lower mean cortisol levels during the LDST (p <0.001), lower peak cortisol (p <0.025), reduced cortisol AUC (p <0.005) and longer time to peak cortisol (p <0.05). Abnormalities were seen in both sexes but were more pronounced in females. Unstimulated adrenal androgen and 17 hydroxyprogesterone concentrations were normal. CONCLUSIONS: Adolescents with CFS have subtle alterations in adrenal function suggesting a reduction in central stimulation of the adrenal glands. The more pronounced effects in females may reflect differential central effects of stress on hypothalamic-pituitary-adrenal axis regulation between the sexes. PMID- 15813609 TI - Leptin and soluble leptin receptor levels in obese children in fasting and satiety states. AB - AIM: To investigate the role of soluble leptin receptors in leptin resistance in obese children. METHODS: Thirty-one obese children (16 boys and 15 girls) with a median age of 12.1 years and 15 age- and sex-matched controls were included in the study. Leptin and soluble leptin receptor levels were measured in fasting and satiety states. RESULTS: Serum leptin levels were significantly higher and soluble leptin receptor levels were significantly lower in obese children compared to controls in fasting and satiety states. In obese children, there was a high and inverse correlation between leptin levels and soluble leptin receptor levels after fasting. Prepubertal obese children had lower leptin and higher soluble leptin, receptor levels compared to pubertal children in both states. CONCLUSION: In this study, being the first to consider both fasting and satiety states, obese children were found to have higher leptin, but lower soluble leptin receptor levels, compared to controls. With these findings, it can be postulated that leptin resistance in obese children originates from a defect of soluble leptin receptor production. PMID- 15813610 TI - Complete androgen insensitivity syndrome caused by the R855H mutation in the androgen receptor gene. AB - Complete androgen insensitivity syndrome (CAIS) is characterized by a completely female phenotype in a 46,XY individual and is caused by mutations in the androgen receptor (AR) gene. A 5 year-old girl presented with bilateral hernia and was noted to have bilateral testes. She had a 46,XY karyotype and was diagnosed with CAIS. To identify the underlying mutation, the exons 2 to 8 of the AR gene were amplified by PCR using sets of known primers and reaction conditions. The results of the mutational analysis for the AR showed the presence of the R855H mutation; her mother was found to be heterozygous and both her 46,XX sister and her aunt had a normal AR gene. This mutation, is the result of a guanine to adenine transition in codon 855 at position 2926 in exon 7 of the AR gene, which causes an alteration of the coding nucleotide triad from CGC to CAC, which subsequently causes the substitution from arginine to histidine in the amino acid sequence of the receptor protein molecule. The same mutation has been reported to cause variable phenotypic expression, which could be explained by the presence of additional co-activating factors modifying the biological activity of the AR. The identification of an AR mutation in a girl with CAIS provides important information, because of the syndrome's genetic heterogeneity. This report emphasizes the fact that genetic determinants outside the coding sequence of the AR can influence the function of the AR protein molecule. Phenotypic expression of the mutation may be used for the construction of maps of functional domains of the AR. PMID- 15813611 TI - Congenital hypothyroidism in association with Caroli's disease and autosomal recessive polycystic kidney disease: patient report. AB - Autosomal recessive polycystic kidney disease (ARPKD) is an important renal disease of childhood. Congenital hypothyroidism has been associated with glomerulocystic kidney disease, but to date no association has been made with ARPKD. To our knowledge this is the first reported case of congenital hypothyroidism in an infant with ARPKD. PMID- 15813612 TI - Autoimmune thyroiditis with associated proteinuria: report of two patients. AB - The association of renal disease and autoimmune thyroid disorders has been reported previously. Renal findings associated with autoimmune thyroiditis present more commonly as proteinuria ranging from mild to nephrotic levels. We report here two adolescent girls with hyperthyroidism associated with transient proteinuria correlated with thyroid hormone levels. They had positive antithyroid peroxidase and antithyroglobulin antibodies. Ultrasonographic and scintigraphic findings of the thyroid gland were consistent with Graves' disease in both. Their renal functions were normal except proteinuria (daily protein excretion of 13.5 mg/m2/h in patient 1 and 11 mg/m2/h in patient 2). When they became euthyroid on antithyroid treatment, proteinuria decreased without associated hematuria and/or hypertension. In conclusion, patients with autoimmune thyroid disease should be assessed for the possibility of proteinuria and the etiological investigation of proteinuria should include evaluation of thyroid functions. PMID- 15813613 TI - A systolic murmur is a common presentation of aortic regurgitation detected by echocardiography. PMID- 15813614 TI - The AbCs of modem management of chronic stable angina pectoris. PMID- 15813615 TI - Aortic stenosis: is it simply a degenerative process or an active atherosclerotic process? AB - Aortic stenosis remains the most common cause of cardiac valve replacement in developed countries. The prevalence of this condition increases with age, and many of the risk factors for coronary artery disease also appear related to the development and progression of aortic stenosis. Recent studies also suggest a relationship between calcium and lipid accumulation in both coronary artery disease and aortic stenosis. Most important, there is growing evidence that aggressive treatment of coronary artery disease risk factors, particularly hyperlipidemia, may influence the progression of aortic stenosis. In this article the current literature is reviewed as it relates to vascular biology, pathogenesis of aortic valve disease, and current and newly emerging management approaches in the care of the patient with aortic stenosis. PMID- 15813616 TI - An update on enhanced external counterpulsation. AB - The development of advanced revascularization techniques has resulted in the growth of a subset of patients with coronary artery disease who are nonrevascularizable and are considered to have refractory angina. Enhanced external counterpulsation (EECP) has been developed for the management of these patients with chronic, refractory disease. Evidence has shown that through improvement of vascular endothelial function and recruitment of collateral vessels, EECP provides many clinical benefits. These patients experience sustained decreases in angina, improvement in exercise time, improved myocardial perfusion, and enhanced quality of life. Furthermore, EECP appears to be safe and effective in the treatment of angina in patients with impaired systolic function and has similar potential in patients with congestive heart failure. PMID- 15813617 TI - Iron and atherosclerosis. AB - Traditional risk factors fail to account for all deaths from coronary artery disease (CAD). Iron can inflict oxidative injury on components of the blood and arterial wall to incite the atherosclerotic process. Medical evidence suggests a role of elevated body iron stores as a risk factor for the development of atherosclerosis. Results of a clinical trial to determine the benefits of reducing body iron stores on cardiovascular mortality are pending. PMID- 15813618 TI - A prospective follow-up of alcohol septal ablation for symptomatic hypertrophic obstructive cardiomyopathy--the Baylor experience (1996-2002). AB - BACKGROUND: There is a paucity of data on the long-term outcome of alcohol septal ablation (ASA) for symptomatic hypertrophic obstructive cardiomyopathy (HOCM). HYPOTHESIS: The study was undertaken to evaluate the longer-term outcome of ASA therapy for symptomatic HOCM. METHODS: In all, 137 patients were enrolled consecutively (1996-1999) and 130 (95%) (74 men, 56 women, aged 51 +/- 17 years) underwent ASA and had serial prospective follow-up for up to 5 years (mean follow up 3.6 +/- 1.4 years). Evaluation included angina (Canadian Cardiovascular Society [CCS] score), dyspnea (New York Heart Association [NYHA] class), duration of exercise on treadmill, and echocardiographic indices. RESULTS: Ethanol (3.5 +/ 1.5 cc), injected into 1.5 +/- 0.6 arteries, induced a mean peak plasma creatine kinase (CK) of 1676 +/- 944 units. Complications of procedures included death 1.5% (2/130), heart block requiring permanent pacemaker 13% (17/130), and coronary dissection 4.4% (6/130). Baseline versus last follow-up visit: NYHA class decreased from 3.0 +/- 0.4 to 1.2 +/- 0.6 (p < 0.01); CCS angina score from 2.0 +/- 0.8 to 0.08 +/- 0.4 (p < 0.01); and duration of exercise increased from 322 +/- 207 to 443 +/- 200 s (p < 0.01). Resting left ventricular outflow tract gradient at baseline versus last follow-up visit showed a decrease from 74 +/- 30 to 4 +/- 13 (p < 0.01), and the dobutamine-provoked gradient of 88 +/- 29 decreased to 21 +/- 21 (p < 0.01) mmHg. All-cause mortality over the duration of follow-up was 7.7% (10) giving an annual rate of 2.1%, and cardiac mortality was 2.3% (3) reflecting an annual rate of 0.6%. CONCLUSIONS: Alcohol septal ablation decreased symptoms and improved exercise performance, indicating that it is an effective procedure for symptomatic HOCM. PMID- 15813619 TI - Role of postoperative use of adrenergic drugs in occurrence of atrial fibrillation after cardiac surgery. AB - BACKGROUND: Use of adrenergic (inotropic and vasopressor) drugs is common after cardiac surgery. HYPOTHESIS: The study was undertaken to evaluate the role of postoperative adrenergic drug use as a predictor of postoperative atrial fibrillation (AF) after cardiac surgery. METHODS: The study population consisted of 199 patients post cardiac surgery. Postoperative adrenergic drug use and the baseline and clinical variables were analyzed as possible predictors of postoperative AF. RESULTS: Of 199 patients, postoperative AF occurred in 59 patients (incidence 30%). The adrenergic drugs were used in 127 (64%) patients. Postoperative AF occurred in 49 of the 127 patients (39%) with and in 10 of the 72 patients (14%) without adrenergic drug use (p < 0.01). By univariate analyses, postoperative adrenergic drug use, age, left ventricular hypertrophy, left atrial size, valve surgery, aortic valve replacement, cross clamp time, bypass time, postoperative ventricular pacing, and hours in intensive care unit were predictors of development of postoperative AF. Atrial pacing was a predictor of freedom from developing AF. By multivariate logistic regression analysis, adrenergic drug use was an independent predictor of postoperative AF (odds ratio [OR] 3.35, 95% confidence interval [CI] 1.38-8.12, p = 0.016). Two other independent predictors were valve surgery (OR 2.88, 95% CI 1.31-6.35, p = 0.002) and age (OR 10.73, 95% CI 10.37-11.10, p = 0.0001). Adrenergic drug use, valve surgery, ventricular pacing, and age were predictors of time duration from surgery to the occurrence of AF. Drugs with predominantly beta1-adrenergic receptor affinity were associated with a higher incidence of postoperative AF (dopamine 44%, dobutamine 41% vs. phenylepherine 20%, p = 0.001). CONCLUSION: Use of adrenergic drugs is an independent predictor of postoperative AF after cardiac surgery. PMID- 15813620 TI - Losartan and end-organ protection--lessons from the RENAAL study. AB - BACKGROUND: The Reduction in ENdpoints with the Angiotensin Antagonist Losartan (RENAAL) study reported that losartan delayed the progression of renal disease in patients with type 2 diabetes and nephropathy. Diabetic or renally impaired patients are at high cardiovascular risk, a risk potentially increased in patients with both conditions. HYPOTHESIS: This post hoc analysis examined whether baseline proteinuria was predictive of cardiovascular outcomes, and whether losartan modifies the risk of cardiovascular outcomes in these patients given its renal-protective effects. METHODS: The RENAAL study compared losartan with placebo (in addition to conventional antihypertensive medications) in type 2 diabetic patients with proteinuria. Morbidity and mortality due to cardiovascular causes were ascertained, and the relationship between baseline proteinuria and cardiovascular outcome was determined. The effect of treatment with losartan was examined using three time-to-event analyses of composite cardiorenal outcomes as described below. RESULTS: Increasing baseline proteinuria was associated with significantly increased risk of myocardial infarction (MI) and all-cause or cardiovascular death, but not stroke. Losartan significantly reduced the risk for the combined endpoint of end-stage renal disease (ESRD), MI, stroke, or death by 21% (p < or = 0.005), irrespective of whether all-cause or cardiovascular death was included in the analysis. In addition, losartan reduced the risk for the composite of ESRD or cardiovascular death by 19.2% (p < 0.05). CONCLUSION: In patients with type 2 diabetes and nephropathy, there is an increased risk of MI and cardiovascular or all-cause mortality. Treatment with losartan is associated with a reduction in proteinuria, a delay in the onset of ESRD, and no increased risk of cardiovascular events in this pre-ESRD population. PMID- 15813621 TI - Multiple coronary fistulae. PMID- 15813622 TI - Exercise-induced left-axis deviation as a marker of proximal LAD stenosis. PMID- 15813623 TI - Platelet function disorder in patients with coronary slow flow. AB - BACKGROUND: Some patients evaluated for chest pain with angiographically normal coronary arteries show coronary slow flow phenomenon (CSFP) on angiography. Slow flow of dye in epicardial coronary arteries is also not an infrequent finding in patients during routine coronary angiography. The precise pathophysiology of CSFP is not known yet. HYPOTHESIS: This study investigates the presence of platelet function disorders in patients with CSFP. METHODS: The patient group included 24 patients with CSFP detected by coronary angiography via the TIMI "frame count" method, and a control group included 23 patients with normal coronary flow. Platelet aggregability induced by use of ristocetin, collagen, and adenosine diphosphate (ADP), was measured from all blood samples in both control and patient groups. RESULTS: The ratio of platelet aggregability increased significantly in patients with CSFP compared with patients with normal coronary flow (ristocetin 57.6 +/- 15 vs. 45.4 +/- 17.1, collagen 62.9 +/- 16.4 vs. 48.9 +/- 25.3, ADP 59.4 +/- 18 vs. 42.4 +/- 15.2, p < 0.05). CONCLUSION: Platelet aggregability is increased in patients with CSFP. PMID- 15813624 TI - Viral load of the human immunodeficiency virus could be an independent risk factor for endothelial dysfunction. AB - BACKGROUND: Recent reports of myocardial infarction in young persons infected with human immunodeficiency virus (HIV) who are receiving protease inhibitor therapy have raised concerns about premature coronary artery disease in this population. However, endothelial dysfunction, hypercoagulability, hypertriglyceridemia, and abnormal coronary artery pathology have been observed in association with HIV infection prior to the availability of protease inhibitor therapy. HYPOTHESIS: The study was undertaken to determine the association between endothelial function, viral load, CD4+ count, and other well-established risk factors for atherosclerosis. METHODS: This prospective, case-controlled study compared viral (HIV) load and the CD4+ T-lymphocyte count and endothelial function in 24 HIV-positive carriers. Brachial artery diameter, HIV viral load, and CD4 count were measured. RESULTS: We found that viral load correlated inversely with endothelial function; the higher the viral load, the worse the endothelial dysfunction (p < 0.005). CONCLUSION: High viral load appears to be associated with endothelial dysfunction in patients with HIV. This preliminary observation supports the infectious theory that viruses may play an important role in the pathogenesis of atherosclerosis. PMID- 15813625 TI - Observational data suggesting there are electrocardiographic abnormalities that strongly suggest the presence of chronic heart failure. AB - Almost nothing is written about the electrocardiographic abnormalities that may suggest the presence of chronic heart failure. Indeed, one often hears that the electrocardiogram does not indicate anything about the function of the ehart. This paper discusses several abnormalities that often predict the presence of chronic heart failure. The information presented is from observations and not from scientific studies. The author urges those who are interested to complete a scientific study based on the premise that such observations may be correct. After all, many scientific studies have been created because of observational data that suggested a certain phenomenon. PMID- 15813626 TI - Albert von Bezold and nervous control of the heart. PMID- 15813627 TI - Girl child today--mother of a nation tomorrow. PMID- 15813628 TI - Comparative study of knowledge, attitude and practices among antenatal care facilities utilizing and non-utilizing women. AB - OBJECTIVE: To compare the knowledge, attitude and practices among antenatal care facilities utilizing and non-utilizing women, aged 15-49 years. METHODS: A Cross sectional survey was conducted between October 2003 and April 2004, including 200 married women in the age range 15-49 years. Knowledge, attitude and practices of women utilizing and not utilizing antenatal care facilities during their previous pregnancy were compared by calculating odds ratios and 95% confidence intervals. P values were obtained by doing chi-square test. RESULTS: Pallor was significantly lower among women utilizing antenatal care (57%) as compared to those who were not (77.6%). (O.R 0.38 95% CI(0.18-0.81) p value 0.02). Tetanus toxoid coverage was higher among women utilizing antenatal care (92%) compared to those who were not (59.2%) (O.R 10.8 95% CI(4.5-26.2). Knowledge about danger signals in pregnancy and realization of the importance of eating a healthy diet during pregnancy was significantly higher among women utilizing antenatal care. CONCLUSION: Lesser prevalence of Anaemia and better tetanus toxoid coverage was seen among women attending antenatal care facilities. Identification of danger signals in pregnancy and recognition of nutritional demands of pregnancy are better understood by women utilizing antenatal care facilities. PMID- 15813629 TI - Diagnostic yield of fast plaque TB test for rapid detection of Mycobacterium in tuberculosis suspects. AB - OBJECTIVE: To compare the diagnostic yield of FAST Plaque TB test with the conventional methods for detection of mycobacterium tuberculosis in sputum of Tuberculosis suspects at Jinnah Postgraduate Medical Center Karachi Pakistan. METHODS: A comparative study of diagnostic yield of FAST Plaque TB test with the culture and ZN staining, conducted from January to June 2004. RESULTS: The study was completed on 48 samples, 31 (64.58%) male and 17 females (35.42%). Half of the cases were sputum positive. Culture positive was in 17 (35.41%) and negative in 28 (58.3%) wereas 3 (6.25%) were contaminated. FAST Plaque TB test was positive in 16 (33.33%) and negative in 32 (66.6%) specimens. Out of 17 culture positive, 2 (11.7%) were negative and in 28 culture negative, 1 (3.57%) specimen was positive for FAST Plaque TB test. Out of 24 smear positive, 11 (45.83%) were negative and in 24 smear negative, 3 (12.5%) were positive, for FAST Plaque TB test. Compared to culture it has sensitivity of 86.23% and specificity of 96.42%, positive predictive value of 93.75% and negative predictive value of 93.1%. CONCLUSION: FAST Plaque TB test is a simple test that can detect viable mycobacterium in 2 days. It has a good sensitivity and specificity. The cost is three times less than the other available tests like PCR. Thus it can be useful in the diagnosis of tuberculosis as an adjunct to sputum microscopy in endemic countries. PMID- 15813630 TI - Spectrum of common childhood skin diseases: a single centre experience. AB - OBJECTIVE: To assess the pattern of childhood skin diseases and to analyze the attitude towards consultation and self-treatment. METHODS: Children with complaints of skin diseases visiting outpatient department of Jinnah Postgraduate Medical Centre and National Institute of Child Health were enrolled in this study during the period of May 2002 to July 2003. A detail performa was filled in, for all the patients. 250 cases were finally analyzed in 15 months duration. The diseases were sub-classified on etiological basis e.g. infectious (bacterial, fungal, viral, parasitic) immune mediated, congenital, allergic and miscellaneous. The children's sex, race, age, duration, history of previous treatment and family and past history for skin or systemic diseases were noted. RESULTS: Of all the patients visiting the outpatient department (OPD), 31% were children with skin diseases. Infectious skin diseases were the commonest (60%). Among the infections, fungal were maximum (20.6%), followed by bacterial diseases (12%). Eczemas constituted 21% of skin diseases and 6.4% children had congenital skin conditions. Previous treatment history was present in 43.6% patients. Antibiotics (topical and/or systemic) were taken by 28.4% of patients; whereas history of steroids (topical and/or systemic) was present in 15.2% of patients. Most children (550) had a normal weight and 13.6% were severely malnourished. CONCLUSION: Skin diseases are fairly common in children especially infectious skin diseases. Eczematous and allergic skin diseases are also frequently encountered in children. Self medication is a common practice. Topical steroids were the most commonly used medications. PMID- 15813631 TI - Knowledge, attitude and practice of parturients regarding Epidural Analgesia for labour in a university hospital in Karachi. AB - OBJECTIVE: To assess the knowledge, sources of knowledge, attitude (fears and misconceptions) regarding Epidural Analgesia (EA) and practices of parturients delivery at Aga Khan University hospital (AKUH). METHODS: A hospital based cross sectional study was conducted at the obstetric unit of AKUH, from November to December 2003. A questionnaire was filled through verbal interviews conducted on 448 parturients fulfilling the inclusion criteria. RESULTS: Seventy six percent of the females were aware of epidural analgesia as a labor pain relieving method. However, only 19% availed EA. About 40% had heard about EA from their obstetricians and 64% from friends or relatives. Twenty even percent believed that EA has detrimental effects on the baby while 9% stated that EA can prolong the duration of labour. CONCLUSION: Majority of the pregnant females delivering at Aga Khan University Hospital were aware of epidural analgesia for labour. However, only a small proportion are availing this service, due to fears and misconceptions. PMID- 15813632 TI - Clinical and echocardiographic characteristics of patients with significant pericardial effusion requiring pericardiocentesis. AB - OBJECTIVE: Clinical and echocardiographic features of significant pericardial effusion (PE) have been reported from the west. Currently there is lack of published data from this part of the world, we reviewed all consecutive cases of significant PE requiring echocardiographic assisted pericardiocentesis to analyze the clinical and echocardiographic features of these patients. METHODS: Forty four consecutive patients who underwent echocardiography assisted pericardiocentesis at the Aga Khan University Hospital (AKUH) between January 1988 and May 2001" re included in this review. RESULTS: Most common presenting symptoms were dyspnea (89%) and fever (36%). Elevated JVP and pulsus paradoxus were documented in 59% and 41% of patients respectively. Sinus tachycardia (75%) and low voltage (34%) were the most common ECG findings. Malignancy (45-51%) and tuberculosis (27%) were among the most frequent causes of PE. One patient died during echocardiography-assisted pericardiocentesis. CONCLUSION: The symptoms and physical findings of haemodynamically significant PE are frequently nonspecific. Transthoracic echocardiography is the gold standard for rapid and confirmatory diagnosis of PE and cardiac tamponade. The most common cause of PE was malignancy followed by tuberculosis. Pericardiocentesis under echocardiographic guidance is a safe and effective treatment for significant PE. PMID- 15813634 TI - Multiple parameters in the diagnosis of ectopic pregnancy. AB - OBJECTIVE: To determine the role of emergency ultrasound examination in the diagnosis and management of ectopic pregnancy. METHODS: A descriptive study done at the Radiology Department, Aga Khan University Hospital, Karachi from January 1999 to December 2000 using observational facts by non-probability convenient sampling method. Seventy one patients who presented with acute symptoms in early pregnancy, were subjected to emergency pelvic ultrasound. Both transabdominal and transvaginal ultrasound were done in 52 patients while in 19 patients only transabdominal ultrasound was performed. Serum Beta hCG levels, of all patients were greater than 1500 iu/ml. RESULTS: Ultrasound correctly identified ectopic pregnancy in 71 patients. Ultrasound diagnosis included live embryo in six patients, adnexal mass with pelvic fluid in 54 patients, extrauterine gestational sac in 4 patients and pelvic fluid without adnexal mass in 7 patients. There was no indeterminate ultrasound scan, resulting in 100% specificity and sensitivity. CONCLUSION: The use of multiple parameters in the diagnosis of ectopic pregnancy resulted in 100% sensitivity and specificity. PMID- 15813633 TI - Cobra snake venom reduces significantly tissue nucleic acid levels in human breast cancer. AB - OBJECTIVE: To look into the feasibility of using cobra snake venom in suppressing breast cancer tissue through inhibition of nucleic acid synthesis. METHODS: Samples of breast cancerous tissue, obtained from Atomic Energy Medical Center, LMCH, Jamshoro, incubated with varying concentrations of venom and without venom (as controls) were assayed for macromolecular (RNA and DNA) levels RESULTS: No. change in the control samples while a prominent and significant fall in nucleic acid contents has been noted in venom treated tissues. Also, maximum effect was observed with 25ug/ml dose. CONCLUSION: Snake venom strongly inhibited the formation of nucleic acids in the breast cancerous tissues. It may bring a fall in cell proliferation, thus there is hope that venom could be used as an anti cancerous drug in the future. PMID- 15813635 TI - Classification of 154 clinical cases of vitamin A deficiency in children (0-15 years) in a tertiary hospital in North West Frontier Province Pakistan. AB - OBJECTIVE: To share our experience of 154 cases of clinical vitamin A deficiency in Pakistani children. METHODS AND RESULTS: The data on age, sex, date of presentation and clinical stage of vitamin A deficiency was recorded. A total of 154 children aged 0-15 years suffering from clinical vitamin A deficiency (xerophthalmia) were recorded. One hundred and twenty three (79.9%) children were 0-6 years with 99 (64.3%) male children. Blinding xerophthalmia (corneal xerosis, corneal ulcers ad keratomalacia) was present in 94 (61%) children. Thirty nine percent children presented in summer, 28% in autumn, 20% in winter and 13% in spring respectively. CONCLUSION: This report indicates that clinical cases of vitamin A deficiency in children do occur in Pakistan. Children under 6 years of age are the most vulnerable age group and there is a seasonal variation in presentation. Both life and sight of these children are at risk. Community based studies may be helpful to identify the magnitude of the problem and possible risk factors at national, provincial and district levels. PMID- 15813636 TI - Biochemical screening for the prediction of preeclampsia. PMID- 15813637 TI - Ovarian primary neuroendocrine carcinoma of non-small cell type: report of an extremely rare neoplasm. AB - Paraffin block sections of a uterus and ovarian mass from a 31 year old female were sent for second opinion to the Pathology Laboratory of Aga Khan University. Histologic examination and immunohistochemical features gave a diagnosis of primary neuroendocrine carcinoma of non-small cell type admixed with benign mucinous cystadenoma. This is a rare tumour with only eight being reported in literature. PMID- 15813638 TI - Systemic lupus erythematosus presenting as hemolytic uremic syndrome: a case report. AB - Associating systemic lupus erythematosus (SLE), with an initial presentation of hemolytic uremic syndrome (HUS) is rare. We report a case of 21-year old Afghani female admitted to our hospital with an initial complaint of high grade fever and diffuse maculopapular rash and swelling of lower limbs. Diagnosis of atypical HUS was established according to the clinical triad of HUS without a veriotoxin producing organism in her stool and the pathological finding compatible to thrombotic microangiopathy. In addition, her symptoms fulfilled the 1982 revised criteria for the classification of SLE. After pulse methylprednisolone, cyclophosphamide and plasmapheresis therapies, her laboratory findings and general condition improved. Unfortunately she was lost to follow up as she decided to return back to Afghanistan. PMID- 15813639 TI - Guillain-Barre syndrome occurring after rabies vaccination. AB - A variety of events are associated with the onset of Guillain-Barre syndrome, including vaccinations and vaccines. These are the swine influenza vaccine, oral poliovirus vaccine and rabies vaccine. Rabies is a uniformly fatal disease. It is preventable if World Health Organization (WHO) guidelines for post exposure treatment (PET) are followed. These include local treatment of wound, passive immunization with rabies immunoglobulins and administration of a efficacious vaccine. Cell culture vaccines are highly immunogenic with fewer side effects, but are costly. For that reason neurotissue vaccines are still widely used in Pakistan, although they are less immunogenic with higher incidence of neuroparalytic complications. We report a case of Guillain-Barre syndrome secondary to sheep brain anti-rabies vaccine in a young boy, who presented with lower limb weakness with total recovery after treatment. PMID- 15813640 TI - Temperature regulation and standardization practices of clinical laboratories in Karachi. AB - OBJECTIVE: To examine the temperature regulation and standardization practices of clinical laboratories in Karachi. METHOD: Forty five clinical laboratories in Karachi were examined for observing the standard protocols for running a lab with particular reference to temperature regulation. A questionnaire to the effect was filled. RESULTS: Among the 45 labs included, the mean complete blood count performed per day was 52+/-47. Only 5 (11%) labs had a temperature reader. Thirty (66.7%) labs had an air-conditioner installed, of which only 24 were found in working condition. Maintenance of instruments was carried out every 67.5+/-.30.6 days. The mean number of haemolyzed samples was 2.3+/-1.7. Control was available in 24(53.3%) labs, which was used daily in only 10 labs. Quantity of blood was the same in all the tubes in only 33(73.3%) labs. CONCLUSION: Commercial laboratories should be properly registered and their quality standardized. PMID- 15813643 TI - Intravenous thrombolysis in acute ischaemic stroke: optimising its use in routine clinical practice. AB - Stroke is a common and important medical problem. Intravenous thrombolysis with alteplase (recombinant tissue plasminogen activator; rtPA) is the only available direct treatment that reduces neurological injury following ischaemic stroke. Strong efficacy data from randomised, controlled trials support the use of intravenous thrombolysis to improve outcomes for patients with acute ischaemic stroke. Numerous studies have provided effectiveness data that demonstrate that intravenous thrombolytic therapy can be given safely outside clinical trial settings. However, effectiveness studies have demonstrated that intravenous thrombolytic therapy is often given despite protocol violations when it is prescribed in routine clinical practice. Protocol violations must be avoided because they are associated with adverse events including higher mortality and increased haemorrhagic complications. Although thrombolytic therapy with alteplase is currently being used in only <10% of patients with acute ischaemic stroke, recent studies demonstrate that quality management efforts can improve both the absolute rate of use as well as the proficiency with which alteplase is administered. Given the complexities inherent in prescribing thrombolysis for patients with acute ischaemic stroke, alteplase should be used by clinicians who are experienced in the diagnosis and management of stroke, working in medical centres that have systems in place to ensure that alteplase is given without protocol violations. PMID- 15813642 TI - Substance P receptor antagonists in psychiatry: rationale for development and therapeutic potential. AB - Increasing evidence suggests that substance P (SP) and its receptor (neurokinin [NK]-1 receptor [NK1R]) might play an important role in the modulation of stress related, affective and/or anxious behaviour. First, SP and NK1R are expressed in brain regions that are involved in stress, fear and affective response (e.g. amygdala, hippocampus, hypothalamus and frontal cortex). Second, the SP content in these areas changes upon application of stressful stimuli. Third, the central administration of SP produces a range of fear-related behaviours. In addition, the SP/NK1R system shows significant spatial overlap with neurotransmitters such as serotonin and noradrenaline (norepinephrine), which are known to be involved in the regulation of stress, mood and anxiety. Therefore, it was hypothesised that blockade of the NK1R might have anxiolytic as well as antidepressant effects. Preclinical studies investigating the effects of genetic or pharmacological NK1R inactivation on animal behaviour in assays relevant to depression and anxiety revealed that the behavioural changes resemble those seen with reference antidepressant or anxiolytic drugs. Furthermore, antagonism or genetic inactivation of the NK1R causes alterations in serotonin and norepinephrine neuronal transmission that are likely to contribute to the antidepressant/anxiolytic activity of NK1R antagonists but that are--at least partially--distinct from those produced by established antidepressant drugs. This underlines the conceivable unique mechanism of action of this new class of compounds. In three independent clinical trials with three different compounds (aprepitant [MK-869], L-759274 and CP-122721), an antidepressant effect of NK1R antagonists could be demonstrated. These results, however, have been challenged by recent failed studies with aprepitant. There are numerous indications from preclinical studies that, in addition to SP and NK1R, other neurokinins and/or neurokinin receptors might also be involved in the modulation of stress-related behaviour and that exclusive blockade of the NK1R might not be sufficient to produce consistent anxiolytic and antidepressant effects. One such candidate is the neurokinin-2 receptor (NK2R), and clinical trials to assess the antidepressant effects of NK2R antagonists are currently underway. Of special interest might also be substances that block more than one receptor type such as NK1/2R antagonists or NK1/2/3R antagonists. These compounds may be more efficacious in antagonising the effects of SP than compounds that only block the NK1R. PMID- 15813644 TI - Psychological symptoms associated with genital herpes virus infections: epidemiology and approaches to management. AB - Genital herpes is a common, chronic, recurrent, viral sexually transmitted infection (STI) occurring worldwide. The first episode may be severe and prolonged, but most recurrences are usually short lived and minor. Although most individuals with this infection have no symptoms, STIs, including genital herpes, often cause psychological and psychosexual morbidity. Unfortunately, the existing data on the psychological symptoms associated with genital herpes have many limitations, including anecdotal reporting, evaluation of small and selected populations, use of varied and sometimes unevaluated questionnaires, and lack of controls. Some individuals with recurrent genital herpes display a range of emotional responses, including depression, anguish, distress, anger, diminution of self-esteem and hostility towards the person believed to be the source of infection. However, it is always important to consider the possibility of pre existing psychopathology. Several retrospective studies have suggested that stress could lead to recurrences of genital herpes infection. However, prospective studies have been unable to demonstrate any relationship between pre existing stress and recurrences. What these studies were able to demonstrate was that stress and recurrences occurred simultaneously, suggesting that perhaps it was the recurrences that were causing stress, rather than the reverse. PMID- 15813645 TI - Social functioning: should it become an endpoint in trials of antidepressants? AB - DSM-IV has recommended use of the Social and Occupational Functioning Scale (SOFAS) as a clinician-rated global assessment scale for measuring social functioning; this scale is analogous to the Clinical Global Impression (CGI) scale traditionally used as a secondary outcome measure in patients with depressive symptoms. However, we believe that health-related quality of life is the most appropriate indicator of social functioning when considering this dimension as an endpoint in clinical trials of antidepressants. As health-related quality of life is a purely subjective measure, patient-rated questionnaires have been found to be most important in this context. In this respect, the Sheehan Disability Scale has been recommended as the most relevant global self-reported assessment of social functioning in trials of antidepressants.A review of questionnaires found that the three most frequently used scales selectively directed at obtaining information about social functioning in trials of antidepressants are the Social Adjustment Scale - Self Report (SAS-SR), the Social Adaptation Self-Evaluation Scale (SASS) and the Short-Form Health Survey (SF-36). However, the number of placebo-controlled trials of antidepressants that have used these scales is still too limited to allow comparisons in terms of responsiveness.Health-related quality of life includes dimensions other than social functioning, e.g. physical health and mental health (including both cognitive and affective problems). The SF-36 includes subscales relating to physical and mental health, which, like the social functioning subscales, are measured in terms of degrees of well being. Another quality-of-life questionnaire, the Quality of Life Enjoyment and Satisfaction Questionnaire (Q LES-Q), covers social, mental and physical problems, in this case measured in terms of degrees of satisfaction. Recently, the Q-LES-Q has been reduced from a comprehensive scale including 60-92 items to a brief version including 15 items. An additional item measures overall life satisfaction. As most of the items in the brief Q-LES-Q include social functioning, the scale can be considered as an alternative to SF-36 or the Sheehan Disability Scale when the focus is on satisfaction with treatment. However, there are insufficient numbers of trials of antidepressants using these questionnaires to allow comparisons. The examples of trials of antidepressants with the SF-36 subscales discussed in this review have mostly involved SSRIs. These trials have demonstrated that although antidepressants improve social functioning compared with placebo over a 6-week treatment period, the endpoint scores are still significantly below the national norms at this point. Only after 12 weeks of therapy are the endpoint scores of the social functioning scales within the limits of the national norms. In relapse prevention trials or in maintenance trials to prevent recurrence of depression, comparisons of social functioning scores with national norms can be important supplementary indicators of the need for treatment. In conclusion, social functioning as part of the health-related concept of the patient-reported quality of-life measure should constitute an endpoint in trials of antidepressants to help clarify the goals of treatment in patients with major depression. In medium- and long-term trials, SF-36 subscales should be used as a supplement to symptom orientated scales. In trials of shorter (6-8 weeks) duration, use of other scales such as the SAS-SR, the Q-LES-Q or the Sheehan Disability Scale should be considered. These scales should be considered as supplementary to each other rather than alternatives; it may be necessary to use more than one of these scales in a trial. PMID- 15813646 TI - HIV-associated neuropathic pain: epidemiology, pathophysiology and management. AB - Peripheral neuropathy is associated with numerous systemic illnesses including HIV infection. Neuropathic pain constitutes approximately 25-50% of all pain clinic visits. Distal symmetrical polyneuropathy (DSP) is the most common form of peripheral neuropathy in individuals with HIV infection. DSP is distinguished from other forms of neuropathy on the basis of history and neurological examination. The pain associated with DSP can be debilitating. Therefore, it is important to diagnose HIV-associated DSP properly and treat the neuropathic pain in order to improve quality of life. We review the clinical manifestations, epidemiology, pathophysiology and management strategies for HIV-associated DSP. PMID- 15813647 TI - Intranasal sumatriptan: in adolescents with migraine. AB - Sumatriptan, a serotonin 5-HT(1B/1D) agonist, constricts cranial blood vessels and inhibits neuroinflammatory processes. A single dose of sumatriptan 10 mg (approved European dosage) was significantly more effective than placebo in achieving headache relief at 1 hour post-dose in a well designed study. Headache relief occurred in significantly more adolescents administered a single dose of intranasal sumatriptan 20 mg (at 1 and 2 hours) and 5 mg (at 2 hours) than placebo (pooled data from two studies). Sustained headache relief (1-24 and 2-24 hours) occurred in significantly more recipients of a single dose of intranasal sumatriptan 20mg and 5mg than placebo (pooled data from two studies). Intranasal sumatriptan was generally well tolerated in adolescent migraineurs (in single episode studies or long term in multiple-episode studies). Taste disturbance occurred more often with intranasal sumatriptan than with placebo [Chart: see text]. PMID- 15813651 TI - Zonisamide: a review of its use in the management of partial seizures in epilepsy. AB - Zonisamide (Zonegran, Excegran) is a new-generation, broad-spectrum antiepileptic drug (AED) currently approved as adjunctive therapy for the treatment of medically refractory partial seizures in adults in the US and as adjunctive therapy or monotherapy in the control of partial and generalised seizures in adults and children in Japan and Korea. Either as adjunctive therapy or monotherapy, zonisamide effectively reduces the frequency of partial seizures, with or without secondary generalisation to tonic-clonic seizures, in adults and children with epilepsy. The drug is generally well tolerated and, additionally, has a favourable pharmacokinetic profile permitting once- or twice-daily administration. Direct head-to-head comparisons with other AEDs would be beneficial in fully defining the place of zonisamide in therapy. In the meantime, adjunctive therapy or monotherapy with zonisamide is a convenient, useful option for the management of partial seizures, including those refractory to other AEDs. PMID- 15813653 TI - Chronic actinic dermatitis in the elderly: recognition and treatment. AB - Chronic actinic dermatitis is a condition involving abnormal cutaneous photosensitivity to UV and, often, visible wavelengths. It is rare, but more common in the elderly than in younger populations, with an estimated prevalence of one in 2000 in the > or = 75-year-old population in Tayside, Dundee, Scotland. It usually presents as a dermatitis that maximally, but not exclusively, affects photo-exposed skin. Investigation to confirm the diagnosis and guide management includes phototesting and patch testing. The mainstay of treatment is education about the condition and advice on sunlight and allergen avoidance. Topical corticosteroids and emollients are always required, at least intermittently, and it is sometimes necessary to resort to systemic immunosuppression, usually with corticosteroids or azathioprine. PMID- 15813652 TI - Cyclo-oxygenase-2 inhibitors: when should they be used in the elderly? AB - Chronic pain in the elderly is frequently a result of arthritic disorders, particularly osteoarthritis. The cyclo-oxygenase (COX)-2 inhibitors are as effective as standard NSAIDs for the relief of pain and for improving function in elderly patients with osteoarthritis and rheumatoid arthritis. COX-2 inhibitors increase the risk of serious gastroduodenal adverse reactions but there is evidence that they carry a lower risk for these adverse effects than standard NSAIDs, except when there is concurrent aspirin use. Since gastroduodenal disorders are the most frequently reported serious adverse effects of NSAIDs and these disorders occur more frequently in the elderly, COX-2 inhibitors offer an alternative to standard NSAIDs in this age group. However, they are not appropriate for many patients with cardiovascular and renal disease. The adverse reaction profile of the COX-2 inhibitors has confirmed the role of the COX-2 enzyme in renal function, salt and water homeostasis and the vascular endothelium. Thus, like standard NSAIDs, COX-2 inhibitors can cause renal failure, hypertension and exacerbation of cardiac failure. Of note is that these disorders are dose related. Thus, there are good reasons to avoid high doses of COX-2 inhibitors in the elderly. Clinical trials indicate that daily doses of rofecoxib 12.5 mg, celecoxib 100-200 mg, valdecoxib 10mg and etoricoxib 60 mg are the minimum effective doses of these agents. Data from the New Zealand Intensive Medicines Monitoring Programme indicate that celecoxib 200 mg/day and rofecoxib 25 mg/day are/were the most commonly prescribed doses and that 6% of patients had taken rofecoxib 50 mg/day for longer than recommended. Recent research indicates that COX-2 inhibitors have a thrombotic potential, especially in high doses and when use is prolonged, and this further limits the extent to which they can be used in the elderly. Important interactions with COX-2 inhibitors in the elderly include those with warfarin, which can result in loss of control of anticoagulation, and those with ACE inhibitors, angiotensin II type 1 receptor antagonists and diuretics, which can result in loss of control of blood pressure and cardiac failure and, in hypovolaemic conditions, renal failure. The clinical significance of an interaction between celecoxib and aspirin to reduce the antiplatelet effect of the latter drug is unknown. Preliminary information from spontaneous reporting systems indicates that there may be differences in the risk of cardiac failure and hypertension between standard NSAIDs and COX-2 inhibitors and between rofecoxib and celecoxib. More formal studies using equivalent doses are needed to test this observation. Use of COX-2 inhibitors may be considered in the elderly to reduce the risk of gastroduodenal complications associated with standard NSAIDs but only when consideration has first been given to use of less toxic medicines as alternatives or supplements, the appropriate dose of the COX-2 inhibitor or standard NSAID, the presence and possible impact of co-morbidities, and the implications of taking COX-2 inhibitors with any concomitant medications. Equally important is regular monitoring of the patient taking a COX-2 inhibitor for efficacy and adverse effects, and ensuring that the patient has a continuing need to keep taking the drug. Close attention also needs to be paid to intercurrent illnesses and new prescriptions that may reduce the safety of the COX-2 inhibitor. A standard NSAID plus a proton pump inhibitor may be equally effective as a COX-2 inhibitor in reducing the risk of gastroduodenal toxicity and if used the same prescribing advice applies. Current knowledge concerning the thrombotic potential of COX-2 inhibitors suggests that this combination, if tolerated, may be preferable to a COX-2 inhibitor, particularly where prolonged use is required. This knowledge also indicates that for patients with or at high risk of ischaemic heart disease or stroke, COX-2 inhibitors are contraindicated. PMID- 15813654 TI - Management of diabetes mellitus medications in the nursing home. AB - Nursing home staff are well aware of the increasing number of residents who experience diabetes mellitus. These residents consume an inordinate amount of resources and often have major disabilities and co-morbidities. Although nonpharmacological therapies, such as consistent carbohydrate intake and increased activity levels, are always indicated in diabetes management, pharmacological therapies are often necessary to prevent the acute complications of diabetes and delay some of the long-term complications. Residents with type 2 diabetes may be managed with oral antidiabetic agents and insulin, whereas residents with type 1 diabetes will always require insulin. Oral antidiabetic agents include insulin secretagogues, which stimulate endogenous insulin secretion and are most effective in leaner persons with type 2 diabetes. Metformin is another oral antidiabetic agent; this decreases inappropriate hepatic glucose release and is most effective in obese residents with high fasting blood glucose levels. The thiazolidinediones, also called glitazones, are insulin sensitisers that enable peripheral tissues to utilise insulin more effectively. The alpha-glucosidase inhibitors delay intestinal absorption of ingested carbohydrates. In addition to oral antidiabetic agents, insulin is frequently used in diabetes management. Insulin is always indicated in type 1 diabetes and is often necessary for residents with type 2 diabetes to optimise glycaemic control. Insulin can be rapid, fast, intermediate or long acting. In addition, basal insulin is now available. These insulins can be combined with each other and, in type 2 diabetes, with oral antidiabetic agents. In order to use pharmacological therapies appropriately, the glycaemic patterns of nursing home residents should be identified, using capillary blood glucose monitoring. Once these patterns have been identified, nonpharmacological therapies can be used, usually in conjunction with the many oral antidiabetic agents and various insulins available, to optimise glycaemic control in each resident. PMID- 15813655 TI - Merkel cell carcinoma of skin: diagnosis and management strategies. AB - Merkel cell carcinoma (MCC) of the skin is a rare form of cutaneous malignancy of neuroendocrine origin with a propensity to affect predominately elderly patients in sun-exposed areas. The tumour has an extremely aggressive growth pattern with the potential to seed into the dermal lymphatics at an early stage as well as spread to nodes and distant sites. Successful outcomes can be achieved with early diagnosis and a multidisciplinary approach to management. Patients with MCC should be carefully staged to exclude distant metastatic disease. Treatment should be approached with both the primary site and the lymph nodes in mind. The primary site should be excised with clear margins, followed by postoperative radiotherapy to the primary site and affected nodes. Addition of chemotherapy to patients perceived to be at high-risk of distant recurrence (e.g. those with involved nodes) has been considered, but evidence supporting this approach is relatively scant. This treatment strategy needs to be approached with caution in the elderly because of the risk of myelosuppression. An aggressive treatment approach is warranted at the first attempt to treat MCC as treatment for recurrent disease is less likely to be successful. PMID- 15813657 TI - Atypical antipsychotics for nursing home patients: a retrospective chart review. AB - BACKGROUND AND OBJECTIVE: To compare the efficacy and safety of risperidone and olanzapine in the treatment of psychotic symptoms and behavioural disturbances in institutionalised patients with dementia and other psychiatric disorders. METHODS: We conducted a retrospective chart review of nursing home patients with psychiatric disorders or dementia in 65 long-term care facilities in New Jersey and Pennsylvania who had received treatment with risperidone or olanzapine. We determined the efficacy of the two antipsychotics for the treatment of psychotic symptoms or behavioural disorders in patients with dementia, the incidence of falls in ambulatory patients, and the incidence of adverse events in the total sample of patients. RESULTS: A total of 289 long-term care patients were included in the analysis. Diagnoses included dementia in 59%, schizophrenia in 20%, bipolar disorder in 8%, schizoaffective disorder in 6% and other diagnoses in 10%. The mean ages were 77 years in patients receiving risperidone and 81 years in patients receiving olanzapine. Risperidone was received by 141 patients and olanzapine by 148 patients. In the 171 patients with dementia, significantly greater improvements in psychotic symptoms and behavioural disturbances were seen in patients receiving risperidone compared with those receiving olanzapine (p < 0.05). In the 222 ambulatory patients, > or = 1 fall was recorded in 19% of patients receiving risperidone and in 38% of patients receiving olanzapine (p = 0.001). The fall rate per month was 0.06 in risperidone recipients and 0.17 in olanzapine recipients (p < 0.001). Adverse events were reported in 6% (9/141) of risperidone-treated patients compared with 34% (42/110) of olanzapine-treated patients (p < 0.001). Adverse events seen only in the olanzapine group were constipation, dry mouth, dysphasia, sedation and dizziness. CONCLUSION: The results of this review indicate that risperidone was more efficacious and better tolerated than olanzapine in the treatment of nursing home patients with psychotic symptoms and behavioural disturbances. PMID- 15813656 TI - Assessing medication adherence in the elderly: which tools to use in clinical practice? AB - Adherence to prescribed medication regimens is difficult for all patients and particularly challenging for the elderly. Medication adherence demands a working relationship between a patient or caregiver and prescriber that values open, honest discussion about medications, i.e. the administration schedule, intended benefits, adverse effects and costs. Although nonadherence to medications may be common among the elderly, fundamental reasons leading to nonadherence vary among patients. Demographic characteristics may help to identify elderly patients who are at risk for nonadherence. Inadequate or marginal health literacy among the elderly is common and warrants assessment. The number of co-morbid conditions and presence of cognitive, vision and/or hearing impairment may predispose the elderly to nonadherence. Similarly, medications themselves may contribute to nonadherence secondary to adverse effects or costs. Especially worrisome is nonadherence to 'less forgiving' drugs that, when missed, may lead to an adverse event (e.g. withdrawal symptoms) or disease exacerbation. Traditional methods for assessing medication adherence are unreliable. Direct questioning at the patient interview may not provide accurate assessments, especially if closed-ended, judgmental questions are posed. Prescription refill records and pill counts often overestimate true adherence rates. However, if elders are asked to describe how they take their medicines (using the Drug Regimen Unassisted Grading Scale or MedTake test tools), adherence problems can be identified in a non-threatening manner. Medication nonadherence should be suspected in elders who experience a decline in functional abilities. Predictors of medication nonadherence include specific disease states, such as cardiovascular diseases and depression. Technological aids to assessing medication adherence are available, but their utility is, thus far, primarily limited to a few research studies. These computerised devices, which assess adherence to oral and inhaled medications, may offer insight into difficult medication management problems. The most practical method of medication adherence assessment for most elderly patients may be through patient or caregiver interview using open-ended, non-threatening and non judgmental questions. PMID- 15813659 TI - Improving survival and reducing toxicity with chemotherapy in advanced non-small cell lung cancer : a realistic goal? AB - The role of chemotherapy in the treatment of advanced non-small cell lung cancer (NSCLC) has been debated over three decades, but it is only relatively recently that chemotherapy has become a standard of care for this disease. In addition to prolonging survival, chemotherapy can palliate distressing symptoms. Concerns that the adverse effects of chemotherapy are likely to outweigh its benefits have largely not been confirmed by quality-of-life data reported among patients with good performance status. Platinum-based chemotherapy regimens in which cisplatin or carboplatin are partnered by a third-generation cytotoxic drug such as gemcitabine, paclitaxel or vinorelbine, have similar activity and efficacy, but differ in adverse effect profiles. Response rates of 30-40% should be expected with median and 1-year survival of 8-10 months and 30-40%, respectively. In the second-line setting chemotherapy with docetaxel has been shown to be significantly superior to best supportive care alone. In a recent trial that compared docetaxel to the novel antifolate, pemetrexed the response rates and survival rates did not differ, but the toxicity profile favored pemetrexed. Overall, these data demonstrate that progress has been made in the use of chemotherapy to improve survival in patients with NSCLC without increasing the incidence of further toxicity. In the past, the potential to survive 1 year was extremely small, whereas now many more patients reach this milestone as well as the 2-year point. However, a plateau has probably been reached with existing cytotoxic drugs and there is a general belief that the next significant advance in the treatment of NSCLC will come from the addition of drugs that target specific molecular pathways in sequence with standard chemotherapy regimens. PMID- 15813658 TI - Cytochrome p450 polymorphisms in geriatric patients: impact on adverse drug reactions--a pilot study. AB - BACKGROUND AND OBJECTIVE: Up to 23% of the population, depending on their ethnic background, has genetically determined differences in the metabolism of drugs by the cytochrome P450 (CYP) enzymes CYP2C9, CYP2C19 and CYP2D6. The aim of this survey was to determine the relationship between genetical polymorphisms in these CYP enzymes and adverse drug reactions (ADRs) in geriatric patients. STUDY DESIGN: In a prospective 6-month cohort study of 243 patients in a geriatric rehabilitation ward, mean age 80.2 +/- 7.7 years, ADRs were identified by intensive monitoring by a pharmacoepidemiological team, consisting of pharmacists and physicians. 125 out of these 243 patients were genotyped cross-sectionally for polymorphisms of CYP2C9, CYP2C19 and CYP2D6 by the TaqMan-polymerase chain reaction. The main outcome measures were the prevalence of genetical polymorphisms and the patients' risk for developing an ADR as related to the genotype. RESULTS: Patients received an average of 14.2 drugs during hospitalisation which led to 251 ADRs in the whole cohort and 149 ADRs in the cross-sectional genotyping study. Genotype frequencies of CYP2C9 enzyme were 25.9% (n = 29) intermediate metabolisers (IMs) and 2.7% (n = 3) poor metabolisers (PMs). For the enzyme CYP2C19, 26.8% (n = 33) IMs and 0.8% (n = 1) PMs were detected. For the enzyme CYP2D6, 24.1% (n = 26) IMs and 3.7% (n = 4) PMs were found in the analysed patient population. In total, 61.6% (n = 77) of genotyped patients experienced mutations in at least one of the three cytochrome enzymes. The ADR rate did not differ significantly between patients with genetic mutations and wild-type genotype patients. Moreover, only eight out of 40 ADRs which were associated with drugs metabolised by CYP2C9, CYP2C19 or CYP2D6 were detected in patients with IM genotype and none in patients with PM genotype. CONCLUSION: In this investigation geriatric patients showed a high rate of ADRs. However, no association between the ADR rate and the patients' genotype could be detected, which most likely was a result of the small number of patient samples analysed. Although prophylactic genotyping would have not prevented ADRs in this pilot study, physicians nevertheless have to be aware of potential genetic mutations in patients with polypharmacy. PMID- 15813660 TI - Selectin antagonists : therapeutic potential in asthma and COPD. AB - Asthma and COPD are chronic inflammatory conditions that affect hundreds of millions of patients worldwide. New therapeutics are desperately needed, especially those that target the underlying causes and prevent disease progression. Although asthma and COPD have distinct etiologies, both are associated with reduced airflow caused by excess infiltration of inflammatory cells into healthy lung tissues. As selectin-mediated adhesion of leukocytes to the vascular endothelium is a key early event in the initiation of the inflammatory response, selectin inhibition is thought to be a good target for therapeutic intervention. Three known selectins are expressed in distinct subsets of cells: P-selectin is presented on the surface of activated platelets and endothelial cells, L-selectin is constitutively expressed on leukocytes, and E selectin synthesis is upregulated in activated endothelial cells. They mediate cell-cell adhesion in the shear flow of the bloodstream via specialized interactions with clusters of oligosaccharides presented on cell surface glycopeptide ligands. The role of selectin-ligand interactions in the inflammatory response has been demonstrated in various animal models, prompting considerable attention from the pharmaceutical industry.Drug discovery efforts have yielded many different classes of selectin inhibitors, including soluble protein ligands, antibodies, oligosaccharides and small molecules. Although many selectin inhibitors have shown activity in preclinical models, clinical progress of selectin-directed therapies has been slow. Early approaches employed carbohydrate-based inhibitors to mimic the natural ligand sialyl Lewis X; however, these compounds proved challenging to develop. Cytel's CY 1503, a complex oligosaccharide, progressed to phase II/III trials for reperfusion injury, but further development was halted when it failed to demonstrate clinical efficacy. Two protein-based selectin inhibitors have reached phase II development. These included Wyeth's recombinant soluble P-selectin ligand, TSI (PSGL-1), which was discontinued after disappointing results in myocardial infarction trials and Protein Design Labs' humanized anti-L-selectin monoclonal antibody, which is currently in development for trauma. Bimosiamose, discovered by Encysive Pharmaceutical and presently being developed by Revotar Biopharmaceuticals, is an 863 g/mol molecular weight dimer with minimal carbohydrate content and is, to date, the leading selectin inhibitor in clinical development. This compound has shown promise in a phase IIa 'proof of concept' trial in patients with asthma, reducing airway recruitment of eosinophils after intravenous administration. Further clinical development of an inhaled formulation is underway. Despite a significant need for new therapeutics, selectin inhibitors have not yet been explored for the treatment of COPD. Bimosiamose represents an important proof of principle, and hopefully continued success will spark renewed interest in selectin-directed therapeutics for respiratory diseases. PMID- 15813661 TI - Severe acute respiratory syndrome: pertinent clinical characteristics and therapy. AB - Severe acute respiratory syndrome (SARS) is a newly emerged infection that is caused by a previously unrecognized virus - a novel coronavirus designated as SARS-associated coronavirus (SARS-CoV). From November 2002 to July 2003 the cumulative number of worldwide cases was >8000, with a mortality rate of close to 10%. The mortality has been higher in older patients and those with co morbidities. SARS has been defined using clinical and epidemiological criteria and cases are considered laboratory-confirmed if SARS coronavirus is isolated, if antibody to SARS coronavirus is detected, or a polymerase chain reaction test by appropriate criteria is positive. At the time of writing (24 May 2004), no specific therapy has been recommended. A variety of treatments have been attempted, but there are no controlled data. Most patients have been treated throughout the illness with broad-spectrum antimicrobials, supplemental oxygen, intravenous fluids, and other supportive measures. Transmission of SARS is facilitated by close contact with patients with symptomatic infection. The majority of cases have been reported among healthcare providers and family members of SARS patients. Since SARS-CoV is contagious, measures for prevention center on avoidance of exposure, and infection control strategies for suspected cases and contacts. This includes standard precautions (hand hygiene), contact precautions (gowns, goggles, gloves) and airborne precautions (negative pressure rooms and high efficiency masks). In light of reports of new cases identified during the winter of 2003-4 in China, it seems possible that SARS will be an important cause of pneumonia in the future, and the screening of outpatients at risk for SARS may become part of the pneumonia evaluation. PMID- 15813662 TI - Management of influenza virus infections with neuraminidase inhibitors: detection, incidence, and implications of drug resistance. AB - Although influenza vaccination remains the primary method for the prevention of influenza, efficacy may be limited by a poor match between the vaccine and circulating strains and the poor response of elderly patients. Hence, there is an important role for antiviral therapy in the management of influenza. While amantadine and rimantadine have been available for the treatment of influenza in some countries for several years, they are only effective against influenza A viruses, they can have neurological and gastrointestinal adverse effects, and resistant virus is rapidly generated. Neuraminidase inhibitors, a new class of drug, are potent and specific inhibitors of all strains of influenza virus, and they have minimal adverse effects. The greatest benefit is seen in those patients presenting <30 hours after development of influenza symptoms, those with severe symptoms or those in high-risk groups. In addition to treatment of the infection, both drugs are effective prophylactically and have been shown to limit spread of infection in close communities, such as families and in nursing homes. No resistant virus strains have been isolated from normal individuals treated with zanamivir. Resistant virus can be isolated from approximately 1% of adults and 5% of paediatric patients with influenza treated with oseltamivir. However, infectivity of mutant viruses is generally compromised. Governments spend millions of dollars on influenza vaccination campaigns; however, once influenza virus is circulating in the community, vaccination cannot limit the spread of disease. A greater promotion of the use of neuraminidase inhibitors for the treatment and prevention of influenza could have a significant impact on limiting its spread. This could result in saving millions of dollars, not only in direct costs associated with medical and hospital care, but also significant savings in indirect costs associated with the loss of productivity at work, school and home environments. For the benefit of all communities, there needs to be a greater awareness of the symptoms of influenza and the efficacy of neuraminidase inhibitors in disease treatment. PMID- 15813663 TI - The disease-modifying effects of twice-weekly oral azithromycin in patients with bronchiectasis. AB - INTRODUCTION: Bronchiectasis is a chronic pulmonary process characterized by recurrent respiratory infections leading to destruction of airways secondary to inflammation. We investigated whether the addition of 6-months' twice-weekly azithromycin to the existing treatment regimen in patients with pulmonary bronchiectasis decreased the number of exacerbations and improved pulmonary function compared with a similar period of time without concurrent azithromycin. METHODS: Thirty patients with high-resolution computed tomography scan-confirmed bronchiectasis were to be recruited. In random order, patients received usual medications for 6 months, and usual medications plus oral azithromycin 500mg twice weekly for 6 months. Patients receiving azithromycin first had a 1-month washout period prior to entering the second phase. Patients recorded weekly peak flow (PF) measurements. Pulmonary function tests (PFTs), 24-hour sputum volume, and needs for intervention with medication or ancillary support were collected at baseline and every 3 months. Exacerbation incidence and sputum volume measurements were compared from baseline to the end of each study phase. RESULTS: Twelve patients were enrolled; 11 were included in the analysis. Owing to randomization, most patients received the azithromycin first, which was fairly well tolerated. PFTs did not change significantly during either study phase and PFs appeared to remain stable during azithromycin therapy and throughout the subsequent control phase. Azithromycin significantly decreased the incidence of exacerbations compared with usual medications (5 vs 16; p = 0.019). Mean 24-hour sputum volume significantly decreased (15% [p = 0.005]) during the active treatment phase, and remained decreased during the control phase (p = 0.028). Subjectively, patients reported increased energy and quality of life while receiving treatment with azithromycin. CONCLUSIONS: The addition of twice-weekly azithromycin significantly decreased the incidence of exacerbation and 24-hour sputum volume and may have stabilized the PFTs and PFs in this 11-patient pilot study. The results of this study justify further investigation of adding azithromycin to the treatment regimens of patients with bronchiectasis for its disease-modifying effects. PMID- 15813664 TI - Comparison of the bronchodilating effects of formoterol and albuterol delivered by hydrofluoroalkane pressurized metered-dose inhaler. AB - OBJECTIVE: To compare the onset of bronchodilation with a new formoterol hydrofluoroalkane (HFA) pressurized metered-dose inhaler (pMDI) with albuterol (salbutamol) HFA pMDI. PATIENTS AND METHODS: Thirty patients with stable mild or moderate asthma (23 using inhaled corticosteroids, mean FEV(1) 82% of predicted, >or=15% reversibility to terbutaline 1mg after 30 minutes) received formoterol HFA (Oxis) 2 x 4.5microg, albuterol HFA (Ventoline) Evohaler) 2 x 100microg, or placebo at three separate visits in this randomized, double-blind, double-dummy, three-way crossover study. FEV(1) was measured before and 3, 10, 20, 30 and 60 minutes after inhalation. Change in FEV(1) at 3 minutes after inhalation was the primary variable. RESULTS: Mean baseline FEV(1) was stable on all study days (range 2.92-2.94L). FEV(1) values at 3 minutes were: formoterol 3.22L (8% increase), albuterol 3.23L (9% increase) and placebo 2.99L (both p < 0.001 vs placebo). Maximum FEV(1) increased similarly with formoterol and albuterol, with no differences observed between the active treatments at any time point. Patients rated treatment effective at 3 minutes in 15 of 30, 19 of 30 and 7 of 30 cases with formoterol, albuterol and placebo, respectively. All treatments were well tolerated. CONCLUSION: In stable, mild, or moderate asthma, formoterol 9microg and albuterol 200microg, both by HFA pMDI, provided equally rapid and effective bronchodilation. PMID- 15813665 TI - Salmeterol/fluticasone propionate versus fluticasone propionate plus montelukast: a cost-effective comparison for asthma. AB - INTRODUCTION AND OBJECTIVE: Asthma, owing to its chronic nature, is associated with a substantial economic burden. Healthcare providers need to compare the cost effectiveness of alternative asthma treatment options to ensure that they obtain the best value for money from the resources they control. The objective of the current study was to compare the cost effectiveness of salmeterol/fluticasone propionate in combination with fluticasone propionate plus montelukast in patients with symptomatic asthma uncontrolled with inhaled corticosteroid (ICS) monotherapy. STUDY DESIGN AND METHODS: Direct healthcare resource data were prospectively collected during a double-blind, randomized, 12-week clinical study of inhaled salmeterol/fluticasone propionate 50/100 microg twice daily (n = 356) and inhaled fluticasone propionate 100 microg twice daily plus oral montelukast 10mg daily (n = 369). Resources were costed in Dutch guilders (NLG) from the perspective of The Netherlands healthcare system using 1999/2000 prices, but have been presented in US dollars and euros. The primary effectiveness measure was the proportion of successfully treated weeks (based on mean morning PEF values). Secondary measures were episode-free days, symptom-free days, and symptom-free nights. RESULTS: Salmeterol/fluticasone propionate was more effective than fluticasone propionate plus montelukast as measured by the proportion of successfully treated weeks mean 63.3% vs 39.0%; median difference 25%; p < 0.001). Salmeterol/fluticasone propionate was also more effective than fluticasone propionate plus montelukast according to the secondary effectiveness measures. The mean total direct daily healthcare costs per patient were 16% higher with fluticasone propionate plus montelukast than with salmeterol/fluticasone propionate mainly due to higher drug costs in the former group (2.25 US dollars vs 1.94; 1.92 euro vs 1.66, respectively; the NLG was fixed against the euro at a rate of 1 euro = NLG2.2 on 31 December 1998; 1 US dollars = NLG1.883, June 2003; 1 US dollars= 0.848 euro, June 2003). Incremental cost-effectiveness analyses showed that salmeterol/fluticasone propionate was dominant over fluticasone propionate plus montelukast and sensitivity analyses showed these results to be robust. CONCLUSION: Salmeterol/fluticasone propionate is a more cost-effective treatment option than fluticasone propionate plus montelukast for patients with symptomatic asthma uncontrolled by ICS. PMID- 15813666 TI - Lucinactant: in neonatal respiratory distress syndrome. AB - Lucinactant, formerly known as KL(4) surfactant, is a novel synthetic lung surfactant containing phospholipids and an engineered peptide, sinapultide, which is designed to mimic the actions of human surfactant protein B. It has been developed for use in the prevention or treatment of respiratory distress syndrome (RDS), a common problem in premature infants, which results from a deficiency or degradation of pulmonary surfactant. Lucinactant is administered intratracheally soon after birth as a replacement surfactant. In the pivotal randomized, double blind, prophylaxis trial in premature infants, the incidence of RDS at 24 hours after birth was significantly lower in lucinactant recipients than in recipients of colfosceril palmitate, a synthetic non-protein-containing surfactant. RDS related mortality at 14 days was significantly lower in lucinactant recipients than in recipients of colfosceril palmitate or beractant, a bovine-derived surfactant. In another randomized, double-blind, prophylaxis trial in premature infants, the rate of survival without bronchopulmonary dysplasia at 28 days of age in lucinactant recipients was not inferior to that in recipients of poractant alfa, a porcine-derived surfactant. Lucinactant was generally well tolerated. Adverse events were transient and related to the administration procedure. There were no differences in the incidences of complications of prematurity between lucinactant and the other surfactants. PMID- 15813670 TI - Spotlight on cefditoren pivoxil in bacterial infections. AB - Cefditoren pivoxil (Spectracef, Meiact) is a third-generation oral cephalosporin with a broad spectrum of activity against pathogens, including both Gram-positive and -negative bacteria, and is stable to hydrolysis by many common beta lactamases. Cefditoren pivoxil is approved for use in the treatment of acute exacerbations of chronic bronchitis (AECB), mild-to-moderate community-acquired pneumonia (CAP), acute maxillary sinusitis, acute pharyngitis/tonsillitis, and uncomplicated skin and skin structure infections (indications may differ between countries). In clinical trials in adults and adolescents, cefditoren pivoxil demonstrated good clinical and bacteriological efficacy in AECB, CAP, acute maxillary sinusitis, acute pharyngitis/tonsillitis, and uncomplicated skin and skin structure infections, and was generally well tolerated. Thus, cefditoren pivoxil is a good option for the treatment of adult and adolescent patients with specific respiratory tract or skin infections, particularly if there is concern about Streptococcus pneumoniae with decreased susceptibility to penicillin, or beta-lactamase-mediated resistance among the common community-acquired pathogens. PMID- 15813671 TI - What is known about autism: genes, brain, and behavior. AB - Autism is a neurodevelopmental disorder of genetic origins, with a heritability of about 90%. Autistic disorder is classed within the broad domain of pervasive developmental disorders (PDD) that also includes Rett syndrome, childhood disintegrative disorder, Asperger syndrome, and PDD not otherwise specified (PDD NOS). Prevalence estimates suggest a rate of 0.1-0.2% for autism and 0.6% for the range of PDD disorders. There is considerable phenotypic heterogeneity within this class of disorders as well as continued debate regarding their clinical boundaries. Autism is the prototypical PDD, and is characterized by impairments in three core domains: social interaction, language development, and patterns of behavior (restricted and stereotyped). Clinical pattern and severity of impairment vary along these dimensions, and the level of cognitive functioning of individuals with autism spans the entire range, from profound mental retardation to superior intellect. There is no single biological or clinical marker for autism, nor is it expected that a single gene is responsible for its expression; as many as 15+ genes may be involved. However, environmental influences are also important, as concordance in monozygotic twins is less than 100% and the phenotypic expression of the disorder varies widely, even within monozygotic twins. Multiple susceptibility factors are being explored using varied methodologies, including genome-wide linkage studies, and family- and case control candidate gene association studies. This paper reviews what is currently known about the genetic and environmental risk factors, neuropathology, and psychopharmacology of autism. Discussion of genetic factors focuses on the findings from linkage and association studies, the results of which have implicated the involvement of nearly every chromosome in the human genome. However, the most consistently replicated linkage findings have been on chromosome 7q, 2q, and 15q. The positive associations from candidate gene studies are largely unreplicated, with the possible exceptions of the GABRB3 and serotonin transporter genes. No single region of the brain or pathophysiological mechanism has yet been identified as being associated with autism. Postmortem findings, animal models, and neuroimaging studies have focused on the cerebellum, frontal cortex, hippocampus, and especially the amygdala. The cerebello-thalamo cortical circuit may also be influential in autism. There is evidence that overall brain size is increased in some individuals with autism. Presently there are no drugs that produce major improvements in the core social or pragmatic language deficits in autism, although several have limited effects on associated behavioral features. The application of new techniques in autism research is being proposed, including the investigation of abnormal regulation of gene expression, proteomics, and the use of MRI and postmortem analysis of the brain. PMID- 15813672 TI - Platelet glycoprotein IIb/IIIa polymorphism and coronary artery disease: implications for clinical practice. AB - Membrane glycoprotein (GP) IIb/IIIa plays a major role in platelet function; indeed it enables stimulated platelets to bind fibrinogen and related adhesive proteins, a process that is considered key in the development of thrombosis. The gene encoding GPIIIa (ITGB3, also known as GP3A) shows a common platelet antigen polymorphism [PL(A1)/PL(A2); expressed by alleles ITGB3*001 and ITGB3*002] that was variably associated with vascular disease. In 1996, the presence of the PL(A2) allele (ITGB3*001) was first reported to increase the risk of coronary heart disease. Shortly after, the interest in this study was increased by the publication of a case report on the death from myocardial infarction of an Olympic athlete who was found to be homozygous for the PL(A2) allele. Overviews of the published studies on the PL(A1)/PL(A2) polymorphism and coronary risk suggest an influence of the PL(A2) allele on the clinical phenotype and the interaction with other environmental factors. In particular, the strongest effect of the ITGB3 PL(A2) allele was expressed on the risk of occlusion after revascularization procedures, mainly after stent implantation, a condition in which platelet activation is more important as compared with other stenotic mechanisms. In the future, the identification of patients who are particularly responsive to GPIIb/IIIa antagonist therapy (e.g. those with the PL(A2) allele) might help to improve the treatment efficacy in this relatively small population. In a mechanism possibly unrelated to its effect on platelet reactivity to aggregating stimuli, the presence of the PL(A2) allele might influence the antiaggregatory effect of platelet inhibitory drugs such as aspirin (acetylsalicylic acid), clopidogrel, and GPIIb/IIIa antagonists. Although interesting, current data does not yet have direct clinical implications for patient risk identification and drug therapy tailoring. Larger studies are necessary to define the role of the PL(A2) allele in more homogeneous groups where platelet GPIIb/IIIa activation might be particularly relevant. PMID- 15813673 TI - Mechanisms of clinical resistance to small molecule tyrosine kinase inhibitors targeting oncogenic tyrosine kinases. AB - A number of highly specific small molecule inhibitors of oncogenic tyrosine kinases have been developed and may potentially improve the treatment of different malignant diseases. However, it became rapidly evident that multiple resistance mechanisms compromise the successful clinical application of these inhibitors, particularly in advanced solid tumors. To develop efficient therapeutic strategies with small molecule inhibitors, one must understand the causes for treatment failure. Three different types of resistance to small molecule inhibitors of oncogenic tyrosine kinases have been observed. The malignant phenotype may be independent of the activity of the target kinase (target-independent resistance). Alternatively, overexpression or mutation of the target kinase can counteract the inhibition of oncogenic tyrosine kinases (target dependent resistance). Finally, alterations of drug transporters or drug metabolizing pathways may block the bioavailability of the tyrosine kinase inhibitors (drug-dependent resistance). This article reviews the current knowledge of clinical resistance to small molecule inhibitors approved for treatment of cancer patients. PMID- 15813674 TI - Pathway proteomics: global and focused approaches. AB - Biological pathways represent the relationships (reactions and interactions) between biological molecules in the context of normal cellular functions and disease mechanisms. Understanding the roles of proteins and signaling pathways expressed within disease, and their link to drug discovery and drug development are central in today's target-driven pharmaceutical processes. This article gives an overview of proteomics strategies, including global expression analysis as well as focused approaches using multidimensional separation by both gel- and liquid-phase techniques linked to mass spectrometry, as applied to two of the pathways involved in inflammatory diseases. In primary human cell studies, our group has annotated and identified thousands of proteins using both electrospray ionization and matrix-assisted laser desorption ionization (MALDI)-sequencing technology. Annotations made from gel images and chromatography fractionation, interfaced to high-end mass spectrometry sequence and structure identity, are cornerstones in cutting-edge protein expression profiling. Regarding phosphorylation mechanisms of kinases, the quantitative stoichiometry can be determined using affinity probe isolations. Another strategy involves micro preparative sample processing, which has been used to analyze single-target phosphoproteins and their relative phospho-stoichiometry. PMID- 15813675 TI - Detection of single nucleotide polymorphisms in the ABCG2 gene in a Dutch population. AB - BACKGROUND: ABCG2 is a drug transporter involved in the protection of tissues by actively transporting toxic substances and xenobiotics out of cells. Cancer cells overexpressing the ABCG2 gene show multidrug resistance to mitoxantrone-, methotrexate-, doxorubicin-, and camptothecin-based anticancer drugs, such as topotecan and SN-38. Large interindividual differences have been shown in oral availability and clearance of drugs that are substrates for ABCG2. Variation in the ABCG2 gene, such as single nucleotide polymorphisms (SNPs), can possibly explain the variability in pharmacokinetics of ABCG2 substrates. AIM: This study was performed to screen for SNPs in the ABCG2 gene to determine the frequencies of currently known and previously unknown SNPs in a Dutch population. METHODS: Blood samples were obtained from 100 healthy volunteers to isolate genomic DNA. PCR amplification was performed, followed by DNA sequencing. The population, of which the ethnicity was 93% Caucasian, consisted of 79 female individuals and 21 males. RESULTS: In total, 19 SNPs were found in the ABCG2 gene, of which 7 were previously unknown. The SNPs G8883A in exon 5 and C44168T in exon 14 cause an amino acid change of R160Q and R575X, respectively. Most of the previously unknown SNPs were found in introns. CONCLUSIONS: The results will be used in future studies to explore the influence of the different SNPs on ABCG2 protein expression, activity, and substrate specificity. In addition, the results can be used to study the effects of genetic polymorphisms in the ABCG2 gene on the pharmacokinetic profile of anticancer drugs. PMID- 15813676 TI - Spotlight on gefitinib in non-small-cell lung cancer. AB - Gefitinib (Iressa), the first commercially available epidermal growth factor receptor-tyrosine kinase (EGFR-TK) inhibitor, is indicated in the management of patients with locally advanced or metastatic non-small-cell lung cancer (NSCLC). However, approved uses differ between countries; in most markets, gefitinib is approved for third-line use only (e.g. the US, Canada and Switzerland), although in some it is approved for both second- and third-line use (e.g. Japan and Australia) and, additionally, in patients considered unsuitable for chemotherapy (e.g. Indonesia and the Philippines). Few third-line treatment options exist for patients with inoperable advanced NSCLC who have failed both docetaxel and platinum-based chemotherapy regimens. Gefitinib represents a significant advance in the treatment of this population; a once-daily oral dosage of 250 mg/day was well tolerated, produced objective tumour responses and disease stabilization, and improved disease-related symptoms and quality of life. It also produced overall survival outcomes that compared favorably with historical outcomes in a similar group of patients treated with three or four different chemotherapy regimens. These findings have been supported by observations from a global compassionate-use program. Ongoing or planned clinical trials are designed to confirm and/or further define the role of the drug in the above and other clinical settings. Preliminary data demonstrate the presence of activating mutations in EGFR-TK among patients whose disease was highly responsive to treatment with gefitinib, although such mutations have not been correlated to all patients who benefit from the drug. Further studies are needed to fully elucidate the clinical implications of EGFR mutations and to identify patients likely to benefit from EGFR-targeted therapy. PMID- 15813677 TI - EGFR pharmacogenomics: the story continues to mutate and evolve. PMID- 15813678 TI - Genes and molecules involved in Aspergillus fumigatus virulence. AB - Aspergillus fumigatus causes a wide range of diseases that include mycotoxicosis, allergic reactions and systemic diseases (invasive aspergillosis) with high mortality rates. Pathogenicity depends on immune status of patients and fungal strain. There is no unique essential virulence factor for development of this fungus in the patient and its virulence appears to be under polygenetic control. The group of molecules and genes associated with the virulence of this fungus includes many cell wall components, such as beta-(1-3)-glucan, galactomannan, galactomannanproteins (Afmp1 and Afmp2), and the chitin synthetases (Chs; chsE and chsG), as well as others. Some genes and molecules have been implicated in evasion from the immune response, such as the rodlets layer (rodA/hyp1 gene) and the conidial melanin-DHN (pksP/alb1 gene). The detoxifying systems for Reactive Oxygen Species (ROS) by catalases (Cat1p and Cat2p) and superoxide dismutases (MnSOD and Cu, ZnSOD), had also been pointed out as essential for virulence. In addition, this fungus produces toxins (14 kDa diffusible substance from conidia, fumigaclavin C, aurasperon C, gliotoxin, helvolic acid, fumagilin, Asp-hemolysin, and ribotoxin Asp fI/mitogilin F/restrictocin), allergens (Asp f1 to Asp f23), and enzymatic proteins as alkaline serin proteases (Alp and Alp2), metalloproteases (Mep), aspartic proteases (Pep and Pep2), dipeptidyl-peptidases (DppIV and DppV), phospholipase C and phospholipase B (Plb1 and Plb2). These toxic substances and enzymes seems to be additive and/or synergistic, decreasing the survival rates of the infected animals due to their direct action on cells or supporting microbial invasion during infection. Adaptation ability to different trophic situations is an essential attribute of most pathogens. To maintain its virulence attributes A. fumigatus requires iron obtaining by hydroxamate type siderophores (ornitin monooxigenase/SidA), phosphorous obtaining (fos1, fos2, and fos3), signal transductional falls that regulate morphogenesis and/or usage of nutrients as nitrogen (rasA, rasB, rhbA), mitogen activated kinases (sakA codified MAP-kinase), AMPc-Pka signal transductional route, as well as others. In addition, they seem to be essential in this field the amino acid biosynthesis (cpcA and homoaconitase/lysF), the activation and expression of some genes at 37 degrees C (Hsp1/Asp f12, cgrA), some molecules and genes that maintain cellular viability (smcA, Prp8, anexins), etc. Conversely, knowledge about relationship between pathogen and immune response of the host has been improved, opening new research possibilities. The involvement of non-professional cells (endothelial, and tracheal and alveolar epithelial cells) and professional cells (natural killer or NK, and dendritic cells) in infection has been also observed. Pathogen Associated Molecular Patterns (PAMP) and Patterns Recognizing Receptors (PRR; as Toll like receptors TLR-2 and TLR-4) could influence inflammatory response and dominant cytokine profile, and consequently Th response to infec tion. Superficial components of fungus and host cell surface receptors driving these phenomena are still unknown, although some molecules already associated with its virulence could also be involved. Sequencing of A. fumigatus genome and study of gene expression during their infective process by using DNA microarray and biochips, promises to improve the knowledge of virulence of this fungus. PMID- 15813679 TI - In vitro activities of voriconazole, fluconazole, itraconazole and amphotericin B against non Candida albicans yeast isolates. AB - Antifungal susceptibility testing was performed on 197 yeast isolates from the BCCM/IHEM biomedical fungi and yeasts collection (Belgian Co-ordinated Collections of Micro-organisms / IPH-Mycology) to study the in vitro activity of voriconazole against fluconazole, itraconazole and amphotericin B. MICs of the four antifungal agents were determined by an adapted NCCLS M27-A microdilution reference method. MIC readings were visually and spectrophotometrically determined. Optical density data were used for calculation of the MIC endpoints. For amphotericin B, the MIC endpoint was defined as the minimal antifungal concentration that exerts 90% inhibition, compared to the control growth. The azoles endpoints were determined at 50% inhibition of growth. The MIC distribution of voriconazole susceptibilities showed that 193 isolates had a MIC < or = 2 microg/ml and 185 a MIC < or = 1 microg/ml. Cross-tabulation of voriconazole, fluconazole, and itraconazole MICs indicated that voriconazole MICs raised with fluconazole and itraconazole MICs. The in vitro data obtained in this study suggest that voriconazole may also be effective treating yeast infection in patients infected with fluconazole or itraconazole resistant isolates. PMID- 15813680 TI - Extracellular enzymatic activity and serotype of Cryptococcus neoformans strains isolated from AIDS patients in Brazil. AB - One hundred and fifty-one Cryptococcus neoformans strains isolated from AIDS patients in Brazil and maintained in the Adolfo Lutz Institute (Sao Paulo, Brazil) were tested for phospholipase, protease and other extracellular enzymatic activities and their serotypes determined. Production of extracellular phospholipase and protease was tested by the agar plate methods. Determination of extracellular enzyme profile of the strains was performed by using the API-ZYM kit system, which can test 19 different enzymes. The serotypes were determined by cell agglutination using the Crypto-check method. Among the 151 strains, 147 were identified as serotype A and four strains were serotype AD. Production of extracellular phospholipase and protease was extensive and observable at early stages of incubation. All of the tested strains were positive for the production of both enzymes. In the API-ZYM tests, more than 90 % of the 151 tested strains were positive for esterase C4 (No. 3), esterase lipase C8 (No. 4), leucine arylamidase (No. 6), phosphatase acid (No. 11), naphtol-AS-BI-phosphohydrolase (No. 12), alpha-glucosidase (No. 16) and beta-glucosidase (No. 17). Differences in enzymatic activities between the Brazilian strains and strains isolated in other countries were observed. The phospholipase, protease and other enzyme activities may play a role in host tissue invasion by C. neoformans. PMID- 15813681 TI - Fungal infection of the feet in soccer players and non-athlete individuals. AB - This study was conducted to evaluate the occurrence of mycoses affecting the feet of soccer players and to compare this results with those in non-athlete individuals of the same age and sex. Initial evaluation consisted of a dermatological examination of the foot in 22 Chinese athletes, 83 Brazilian athletes and 24 Brazilian non-athletes. Scales of plantar skin, interdigital and subungual areas of the foot were collected for mycological examination (direct and culture). Nail clippings were obtained for histopathologic analysis. Tinea pedis was diagnosed more frequently among the non-athlete individuals. None of the Chinese athletes had tinea pedis alone. However, in this group onychomycosis was frequently higher when compared to the other groups. The fungal microbiota comprised Trichophyton rubrum (40%), Trichophyton mentagrophytes (36.4%) and Candida spp (20%). Candida spp was isolated only from Brazilian athletes. Results obtained with KOH wet mounts agreed with the results obtained in culture and with histopathologic examinations (50.5% vs 40.9%). The frequency of tinea pedis among soccer players was lower than the other groups in this study, possibly due to health education and professional feet care. PMID- 15813682 TI - [An ELISA test for the study of the therapeutic evolution of chromoblastomycosis by Cladophialophora carrionii in the endemic area of Falcon State, Venezuela]. AB - The purpose of this research was to evaluate an ELISA indirect method in patients with chromoblastomycosis caused by Cladophialophora carrionii. Samples collected before, during and postreatment with ajoene or itraconazole, and those from apparently healthy people from the endemic area were evaluated with the ELISA test. 94 individuals were studied, 10 with chromoblastomycosis, and 84 apparently healthy subjects. All of them were evaluated by clinical-dermatological examinations. On those with lesions suggestive of chromoblastomycosis, mycological studies were carried out to confirm the disease. This approach was repeated during and at the end of therapy. Five patients with lesions < or = 5 cms were treated with ajoene and five with lesions > 5 cms, received itraconazole. Mycological cure (60%) was similar in both groups of patients and persisted three months after therapy. One hundred and fourteen sera were analyzed by ELISA, 30 from 10 patients with chromoblastomycosis, before, during and postreatment and 84 from apparently healthy people, using a somatic antigen of C. carrionii (AgSPP). All patients with chromoblastomycosis were positive before treatment, two became negative on day 45 of treatment and a total of six patients were negative three months post-treatment. All sera from apparently healthy individuals were negative. The sensitivity and specificity was 100% and 98.9%, respectively. The relationship between clinical-mycological studies and the ELISA assay was 100% before and after treatment. In summary, ELISA could be a valuable tool for the diagnosis and evolution of the therapeutic efficacy in patients with chromomycosis (C. carrionii). The use of an ELISA test is therefore highly recommended to establish remission criteria in chromoblastomycosis caused by C. carrionii. PMID- 15813683 TI - Immunohistochemical characterization of the cellular infiltrate in Jorge Lobo's disease. AB - Few studies have been conducted to evaluate the cellular composition of the granulomatous lesions induced by Lacazia loboi. Thus, the objective of the present study was to characterize the mononuclear cell population present in cutaneous lesions obtained from 15 patients with Jorge Lobo's disease. Histological sections were stained with hematoxylin-eosin and methenamine silver and the following mononuclear cells were identified by immunohistochemistry: T lymphocytes (CD3+), helper T lymphocytes (CD4+), cytotoxic T lymphocytes (CD8+), B lymphocytes (CD20+), plasma cells (CD79+), natural killer cells (CD57+) and histiocytes (CD68+). This study showed that the inflammatory infiltrate mainly consists of histiocytes and multinucleated giant cells, in addition to the presence of a large number of fungal cells. The identified inflammatory cells showed the following frequency: CD68+ histiocytes > CD3+ T lymphocytes > CD4+ T > CD8+ T lymphocytes > CD57+ natural killer cells > CD79+ plasma cells > CD20+ B lymphocytes. Based on the findings of a large number of fungal cells in the infected tissues and the disorganized cell arrangement in the granuloma, we hypothesize that patients with Jorge Lobo's disease present immunoregulatory disturbances, which are likely to be specific and perhaps responsible for the lack of containment of the pathogen. PMID- 15813684 TI - [New fungal contaminants of food products in Argentina]. AB - Five fungal species were isolated and identified in food products: Ascotricha chartarum, Leptosphaerulina argentinensis, Veronaea coprophila, Scolecobasidium constrictum and Coremiella cubispora. A. chartarum was isolated from paper bag containing sugar and the other four from tomato sauce. Except for L. argentinensis, the other four were new reports and the five species were isolated for the first time in Argentina. PMID- 15813685 TI - [Tinea capitis in an adult. Case report]. AB - A tinea capitis and tinea faciei case in an elderly female patient with diabetes mellitus type 2 is presented. Her dermatological illness began on her face and later disseminated to her scalp. She received multiple medications which aggravated her condition. The diagnosis was established by means of direct examination and the isolation of Trichophyton tonsurans in culture. Treatment with 200 mg daily dose of itraconazol for two months resulted in clinical and mycological cure. PMID- 15813686 TI - Isolation, identification and antifungal susceptibility of lemon pathogenic and non pathogenic fungi. AB - Numerous species of filamentous fungi were isolated from lemon on different plantations in the province of Tucuman, Argentina. The techniques suggested by the Subcommittee of Antifungal Susceptibility of the National Committee for Clinical Laboratory Standards, (USA) were adapted. The effect of three different concentrations of the fungicides imazalil, guazatine, SOPP and thiabendazole on the fungi Fusarium oxysporum, Fusarium moniliforme, Aspergillus niger, Aspergillus flavus, Aspergillus clavatus, Geotrichum candidum, Rhizopus sp, Penicillium sp, Penicillium digitatum and Mucor sp were studied. All the tested strains were resistant to thiabendazole. We assayed a mixture of SOPP (5%), guazatine (350 ppm) and imazalil (100 ppm), which showed a synergic effect on Rhizopus sp. Mucor sp was the only fungus resistant to the four fungicides tested as well as to the above mentioned mixture. PMID- 15813687 TI - [Clinical cases in Medical Mycology. Case No. 14]. PMID- 15813688 TI - [Clinical cases in Medical Mycology. Case No. 15]. PMID- 15813689 TI - [Emerging mycotoxins: introduction]. AB - Mycotoxins are a diverse group of secondary metabolites produced by moulds. They have a wide range of toxicological effects both in humans and animals. Nowadays about 200 of these metabolites have been described, but only a few of them may be considered as important from the agricultural point of view. In this publication, some aspects such as mycotoxin producing species, analytical methods, natural occurrence, preventive and detoxification techniques and regulations of these kind of mycotoxins will be developed by different authors involved in these research issues. PMID- 15813690 TI - [Mycotoxin producing fungi]. AB - Mycotoxins are relatively small molecules characterized by a diversity of chemical structure and a diversity of biological activity. They are often genotypically specific for a group of species, but the same compound can also be formed by fungi belonging to different genera. Most of the mycotoxins known have been recognized as metabolic products of Aspergillus, Penicillium and Fusarium species. This review will be focused on aflatoxins, ochratoxins and fumonisins because of their hazard to animal and human health. The production of these mycotoxins have been usually associated with a small number of species but some recent studies have reported the production of these mycotoxins by some other species. These results show that mycotoxin production is broader than is normally thought, so the possibility can not be ruled out that new species may be a new source of unexpected mycotoxins in their natural substrates. PMID- 15813691 TI - [Emerging mycotoxin control. Current limits and regulations]. AB - Emerging mycotoxins are of a great importance due to their high occurrence in foods and feeds and their implication in pathologies. There is a need to control them both in pre- and post-harvest. However, the increasing drawbacks of chemical decontamination methods, makes the use of preventive methods more attractive. These are based on avoiding growth of mycotoxigenic fungi and mycotoxin accumulation in these substrates (by using reduced aw levels, and modifying other storage conditions) and the early detection of these mycotoxins by using simple and fast analysis techniques. At the same time, it is important to know the effect that industrial processing has on mycotoxin concentration in foods and feeds, because it may reduce the toxin content. Finally, maximum limits of mycotoxin concentration should be established for both foods and feeds. PMID- 15813692 TI - [Thougths at the doors of Third Millennium]. PMID- 15813693 TI - [Morphology of severe mycoses. Role of the pathologist in the diagnosis]. AB - Mycoses are diseases caused by fungi. To be regarded as pathogenic, a fungus has to be able to complete a number of steps in order to initiate infection. Although culture remains the gold standard to diagnose the causative agent, presumptive identification of these fungi in histologic slides is of great value because invasion is only demonstrated in tissue sections. When culture is not available, histopathologic examination may be the sole source of information about the nature of the infection. To make a diagnosis, the pathologist needs to be familiar with the morphologic appearance of the different types of fungi, and has also to be aware of the limitations in their identification. The purpose of this work is to review the current problems on this subject, specially referring to the most frequent severe mycoses of our environment. PMID- 15813694 TI - Current knowledge on pathogenesis and immunodiagnosis of paracoccidioidomycosis. AB - Host-parasite relationship and immunodiagnostic testing in paracoccidioidomycosis have been extensively investigated in recent years. We review the major advances in the understanding of pathogenesis of the disease with emphasis on the sequential steps in granuloma formation and the envolvement of immunological mechanisms in host defenses against the parasite. In addition, the several immunodiagnostic tests used for diagnosis and in the follow-up of patients are commented upon and critically analysed. PMID- 15813695 TI - Comparison of DNA-based typing methods to assess genetic diversity and relatedness among Candida albicans clinical isolates. AB - Three serial isolates of Candida albicans were obtained from each of five HIV infected patients with recurrent oropharyngeal candidiasis from the same geographical area. Isolates from one patient remained susceptible to fluconazole whereas serial isolates from the other four patients showed decreasing susceptibilities to the drug. Strain identity was investigated by pulse-field gel electrophoretic (PFGE) separation of chromosomes, restriction fragment length polymorphism (RFLP) of chromosomal DNA, Southern blot analysis with the moderately repetitive probe Ca3 of the materials present in the RFLP gels after transfer to nylon membranes, and random amplification of polymorphic DNA (RAPD). All techniques were able to group isolates obtained from the same patient. Techniques resulting in more complex banding profiles exhibited increased discriminatory power allowing detection of strain variants. Methods resulting in less complex banding patterns, especially Southern hybridization of SfiI digested chromosomal DNA with the moderately repetitive probe Ca3, were more helpful to determine isogenicity among isolates obtained from the same patient. The combination of results from methods with high discriminatory power (to maximize detection of strain variants) and methods resulting in less complex banding patterns (to allow determination of isogenic isolates) should facilitate the delineation of the epidemiology of C. albicans infection. PMID- 15813696 TI - Effects of probiotic bacteria on humoral immunity to Candida albicans in immunodeficient bg/bg-nu/nu and bg/bg-nu/+ mice. AB - Germfree beige-nude ( bg/bg-nu/nu) and beige-heterozygous ( bg/bg-nu/+) mice were colonized with a pure culture of Candida albicans or with a probiotic bacterium (Lactobacillus acidophilus, Lactobacillus reuteri, Lactobacillus casei, or Bifidobacterium infantis). Probiotic-colonized mice were subsequently challenged orally with C. albicans. The effect of prior colonization with probiotic bacteria on the antibody responses of the immunodeficient mice to alimentary tract colonization with C. albicans was compared to the antibody responses of the gnotobiotic mice colonized only with C. albicans. This study demonstrated that, although the probiotic bacteria did not induce a vigorous antibody response to their own antigens, they altered the antibody responses of mice to C. albicans. In T cell competent bg/bg-nu/+mice, B. infantis enhanced and focused IgG1, IgG2A, and IgA responses to C. albicans antigens. Some of the probiotic bacteria also enhanced the IgG1 and IgG2A antibody responses of bg/bg-nu/nu mice to C. albicans antigens. This study not only shows the value of gnotobiotic animal models in demonstrating that probiotic bacteria can affect the capacity of mice to form antibodies to C. albicans, but it also points out their usefulness in comparing the capacity of different probiotic bacteria to produce beneficial health effects in mice. PMID- 15813697 TI - In vitro susceptibilities of Candida and Aspergillus species to Melaleuca alternafolia (tea tree) oil. AB - Candida species are an important cause of opportunistic infection in the oral cavity of immunocompromised patients, especially HIV infected patients. Melaleuca oil obtained commercially was investigated since it is known to have broad antifungal properties. The in-vitro susceptibilities of Aspergillus and susceptible and resistant Candida species were performed utilizing serial dilutions in microtiter plates with Sabouraud dextrose agar and the commercial preparation of Melaleuca. As a comparator, in vitro susceptibilities to amphotericin B and fluconazole were also determined using the broth microdilution technique. The results demonstrate that Melaleuca inhibited the Candida species. However, the growth of Aspergillus was not inhibited at the concentrations tested. Thus, preparations containing Melaleuca alternafolia may be a useful alternative for superficial candidal infections. In fact, it may be a useful alternative regimen for advanced HIV-positive patients with oropharyngeal candidiasis refractory to fluconazole. However, controlled clinical studies to evaluate its efficacy are still needed. PMID- 15813698 TI - Extracellular ABTS-oxidizing activity of autochthonous fungal strains from Argentina in solid medium. AB - The screening for extracellular oxidases and peroxidases from autochthonous filamentous fungi isolated from different substrates is an important step towards the detection of extracellular fungal oxidative systems. Thirty-one autochthonous fungal strains from Argentina, belonging to different ecophysiological and taxonomic groups, were plate-screened for their ability to produce extracellular oxidoreductases. Modified Kirk solid medium containing the chromogen 2,2-azino bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) was used to determine the presence of this extracellular activity. The fungi tested were grouped according to the colour intensity of the modified Kirk medium in: a) species without extracellular ABTS-oxidizing activity; b) species with low extracellular ABTS oxidizing activity; c) species with moderate extracellular ABTS-oxidizing activity; d) species with high extracellular ABTS-oxidizing activity. The assay revealed extracellular ABTS-oxidizing activity in 90% of the strains tested. All species of Basidiomycetes used exhibited ABTS-oxidizing activity, except Laeticorticium roseum. Aspergillus terreus and Epicoccum purpurascens (Deuteromycetes) did not show extracellular oxidative activity on ABTS. Agrocybe aegerita, Amauroderma boleticeum, Cladosporium cladosporioides, Coriolopsis rigida, Grammothele subargentea, Graphium putredinis, Hexagona hydnoides, Hexagona papyraceae, Loweporus lividus, Peniophora albobadia, Phellinus everhartii, Phellinus gilvus; Phellinus linteus; Pleurotus laciniatocrenatus, Pycnoporus sanguineus, Rigidoporus ulmarius, Steccherinum rawakense, Talaromyces helicus, Trametes elegans, Trametes pavonia, Trametes villosa and Trichaptum sector are reported here for the first time as species capable of producing ABTS oxidizing extracellular oxidorreductases. PMID- 15813699 TI - A human corneal equivalent constructed from SV40-immortalised corneal cell lines. AB - Within the last decade, extensive research in the field of tissue and organ engineering has focused on the development of in vitro models of the cornea. The use of organotypic, three-dimensional corneal equivalents has several advantages over simple monolayer cultures. The aim of this study was to develop a corneal equivalent model composed of the same cell types as in the natural human tissue, but by using immortalised cell lines to ensure reproducibility and to minimise product variation. We report our success in the establishment of an SV40 immortalised human corneal keratocyte cell line (designated HCK). A collagen matrix, built up with these cells, displayed the morphological characteristics of the human stromal tissue and served as a biomatrix for the immortalised human corneal epithelial and endothelial cells. Histological cross-sections of the whole-cornea equivalents resemble human corneas in tissue structure. This organotypic in vitro model may serve as a research tool for the ophthalmic science community, as well as a model system for testing for eye irritancy and drug efficacy. PMID- 15813700 TI - Subcellular localization and functional expression of the glycerol uptake protein 1 (GUP1) of Saccharomyces cerevisiae tagged with green fluorescent protein. AB - GFP (green fluorescent protein) from Aequorea victoria was used as an in vivo reporter protein when fused to the N- and C-termini of the glycerol uptake protein 1 (Gup1p) of Saccharomyces cerevisiae. The subcellular localization and functional expression of biologically active Gup1-GFP chimaeras was monitored by confocal laser scanning and electron microscopy, thus supplying the first study of GUP1 dynamics in live yeast cells. The Gup1p tagged with GFP is a functional glycerol transporter localized at the plasma membrane and endoplasmic reticulum levels of induced cells. The factors involved in proper localization and turnover of Gup1p were revealed by expression of the Gup1p-GFP fusion protein in a set of strains bearing mutations in specific steps of the secretory and endocytic pathways. The chimaerical protein was targeted to the plasma membrane through a Sec6-dependent process; on treatment with glucose, it was endocytosed through END3 and targeted for degradation in the vacuole. Gup1p belongs to the list of yeast proteins rapidly down-regulated by changing the carbon source in the culture medium, in agreement with the concept that post-translational modifications triggered by glucose affect proteins of peripheral functions. The immunoelectron microscopy assays of cells expressing either Gup1-GFP or GFP-Gup1 fusions suggested the Gup1p membrane topology: the N-terminus lies in the periplasmic space, whereas its C-terminal tail has an intracellular location. An extra cytosolic location of the N-terminal tail is not generally predicted or determined in yeast membrane transporters. PMID- 15813701 TI - Radiation-induced cell death: importance of lysosomal destabilization. AB - The mechanisms involved in radiation-induced cellular injury and death remain incompletely understood. In addition to the direct formation of highly reactive hydroxyl radicals (HO*) by radiolysis of water, oxidative stress events in the cytoplasm due to formation of H2O2 may also be important. Since the major pool of low-mass redox-active intracellular iron seems to reside within lysosomes, arising from the continuous intralysosomal autophagocytotic degradation of ferruginous materials, formation of H2O2 inside and outside these organelles may cause lysosomal labilization with release to the cytosol of lytic enzymes and low mass iron. If of limited magnitude, such release may induce 'reparative autophagocytosis', causing additional accumulation of redox-active iron within the lysosomal compartment. We have used radio-resistant histiocytic lymphoma (J774) cells to assess the importance of intralysosomal iron and lysosomal rupture in radiation-induced cellular injury. We found that a 40 Gy radiation dose increased the 'loose' iron content of the (still viable) cells approx. 5 fold when assayed 24 h later. Cytochemical staining revealed that most redox active iron was within the lysosomes. The increase of intralysosomal iron was associated with 'reparative autophagocytosis', and sensitized cells to lysosomal rupture and consequent apoptotic/necrotic death following a second, much lower dose of radiation (20 Gy) 24 h after the first one. A high-molecular-mass derivative of desferrioxamine, which specifically localizes intralysosomally following endocytic uptake, added to the culture medium before either the first or the second dose of radiation, stabilized lysosomes and largely prevented cell death. These observations may provide a biological rationale for fractionated radiation. PMID- 15813702 TI - Spectral and catalytic properties of aryl-alcohol oxidase, a fungal flavoenzyme acting on polyunsaturated alcohols. AB - Spectral and catalytic properties of the flavoenzyme AAO (aryl-alcohol oxidase) from Pleurotus eryngii were investigated using recombinant enzyme. Unlike most flavoprotein oxidases, AAO does not thermodynamically stabilize a flavin semiquinone radical and forms no sulphite adduct. AAO catalyses the oxidative dehydrogenation of a wide range of unsaturated primary alcohols with hydrogen peroxide production. This differentiates the enzyme from VAO (vanillyl-alcohol oxidase), which is specific for phenolic compounds. Moreover, AAO is optimally active in the pH range of 5-6, whereas VAO has an optimum at pH 10. Kinetic studies showed that AAO is most active with p-anisyl alcohol and 2,4-hexadien-1 ol. AAO converts m- and p-chlorinated benzyl alcohols at a similar rate as it does benzyl alcohol, but introduction of a p-methoxy substituent in benzyl alcohol increases the reaction rate approx. 5-fold. AAO also exhibits low activity on aromatic aldehydes. 19F NMR analysis showed that fluorinated benzaldehydes are converted into the corresponding benzoic acids. Inhibition studies revealed that the AAO active site can bind a wide range of aromatic ligands, chavicol (4-allylphenol) and p-anisic (4-methoxybenzoic) acid being the best competitive inhibitors. Uncompetitive inhibition was observed with 4 methoxybenzylamine. The properties described above render AAO a unique oxidase. The possible mechanism of AAO binding and oxidation of substrates is discussed in the light of the results of the inhibition and kinetic studies. PMID- 15813703 TI - The N domain of somatic angiotensin-converting enzyme negatively regulates ectodomain shedding and catalytic activity. AB - sACE (somatic angiotensin-converting enzyme) consists of two homologous, N and C domains, whereas the testis isoenzyme [tACE (testis ACE)] consists of a single C domain. Both isoenzymes are shed from the cell surface by a sheddase activity, although sACE is shed much less efficiently than tACE. We hypothesize that the N domain of sACE plays a regulatory role, by occluding a recognition motif on the C domain required for ectodomain shedding and by influencing the catalytic efficiency. To test this, we constructed two mutants: CNdom-ACE and CCdom-ACE. CNdom-ACE was shed less efficiently than sACE, whereas CCdom-ACE was shed as efficiently as tACE. Notably, cleavage occurred both within the stalk and the interdomain bridge in both mutants, suggesting that a sheddase recognition motif resides within the C domain and is capable of directly cleaving at both positions. Analysis of the catalytic properties of the mutants and comparison with sACE and tACE revealed that the k(cat) for sACE and CNdom-ACE was less than or equal to the sum of the kcat values for tACE and the N-domain, suggesting negative co-operativity, whereas the kcat value for the CCdom-ACE suggested positive co-operativity between the two domains. Taken together, the results provide support for (i) the existence of a sheddase recognition motif in the C domain and (ii) molecular flexibility of the N and C domains in sACE, resulting in occlusion of the C-domain recognition motif by the N domain as well as close contact of the two domains during hydrolysis of peptide substrates. PMID- 15813704 TI - Regulation of ecdysteroid signalling during Drosophila development: identification, characterization and modelling of ecdysone oxidase, an enzyme involved in control of ligand concentration. AB - The steroidal moulting hormones (ecdysteroids) mediate developmental transitions in insects, and their regulation is mainly controlled by the production and inactivation of these steroid hormones at the appropriate developmental times. One route of metabolism of ecdysteroids in insects involves EO (ecdysone oxidase) catalysed conversion into 3-dehydroecdysteroid, which undergoes reduction to the corresponding 3-epiecdysteroid. By a twin-stranded bioinformatics approach, employing both phylogenomics and model structure-based analysis, we first predicted that DmEO (the EO of Drosophila melanogaster) corresponds to the protein product of gene CG9504. When CG9504 was expressed in COS7 cells, significant conversion of ecdysone into 3-dehydroecdysone was observed. Quantitative PCR and enzyme assay showed that DmEO was mainly expressed in the midgut during the late instars at a time corresponding to a hormone titre peak. DmEO shares only 27% amino acid sequence identity with Spodoptera littoralis (Lepidoptera) EO, yet key substrate-binding residues are well conserved. A model of DmEO is consistent with an inability to catalyse reaction of cholesterol derivatives. The significance of DmEO in ligand activation is discussed in relation to new evidence suggesting that 3-dehydro- and 3-epiecdysteroids may be functionally active as ligands in a novel, atypical ecdysteroid signalling pathway involving the Drosophila orphan nuclear receptor, DHR38, rather than being merely hormone inactivation products. PMID- 15813705 TI - Characterization of recombinant Saccharomyces cerevisiae telomerase core enzyme purified from yeast. AB - Telomerase is a cellular reverse transcriptase that elongates the single-stranded chromosome ends and oligonucleotides in vivo and in vitro. In Saccharomyces cerevisiae, Est2p (telomerase catalytic subunit) and Tlc1 (telomerase RNA template subunit) constitute the telomerase core complex. We co-overexpressed GST (glutathione S-transferase)-Est2p and Tlc1 in S. cerevisiae, and reconstituted the telomerase activity. The GST-Est2p-Tlc1 complex was partially purified by ammonium sulphate fractionation and affinity chromatography on glutathione beads, and the partially purified telomerase did not contain the other two subunits of the telomerase holoenzyme, Est1p and Est3p. The purified recombinant GST-Est2p Tlc1 telomerase core complex could specifically add nucleotides on to the single stranded TG(1-3) primer in a processive manner, but could not translocate to synthesize more than one telomeric repeat. The purified telomerase core complex exhibited different activities when primers were paired with the Tlc1 template at different positions. The procedure of reconstitution and purification of telomerase core enzyme that we have developed now allows for further mechanistic studies of the functions of other subunits of the telomerase holoenzyme as well as other telomerase regulation proteins. PMID- 15813706 TI - Membrane binding of beta2-glycoprotein I can be described by a two-state reaction model: an atomic force microscopy and surface plasmon resonance study. AB - Complexes formed between beta2GPI (beta2-glycoprotein I), a human plasma protein, and biological membranes are considered to be targets of macrophages and antiphospholipid autoantibodies involved in autoimmune diseases, such as antiphospholipid syndrome or systemic lupus erythematosus. The positively charged lysine-rich fifth domain of beta2GPI facilitates its interaction with phospholipid membranes containing acidic phospholipids, which normally become exposed by apoptotic processes. In the present study, atomic force microscopy was applied to visualize the binding of beta2GPI to a mixed phospholipid model membrane at physiological ionic strength. On supported lipid bilayers the formation of supramolecular assemblies of the protein with a height of approx. 3.3 nm was observed, suggesting a lateral agglomeration of beta2GPI. Detailed analysis of kinetic constants using surface plasmon resonance revealed that the binding can be described by a two-state reaction model, i.e. a very fast interaction step, depending on the content of acidic phospholipids in the bilayer, and a second step with significantly lower k(on) and k(off) values. Taken together, our results suggest a biphasic interaction mechanism: a fast step of beta2GPI binding to negatively charged lipids, mainly based on electrostatic interactions, and a slower phase of agglomeration of the protein on the bilayer surface accompanied by a protein-induced rigidification of the membrane, as revealed by electron paramagnetic resonance. PMID- 15813707 TI - Structure-function relationship in annexin A13, the founder member of the vertebrate family of annexins. AB - Annexin A13 is considered the original progenitor of the 11 other members of vertebrate annexins, a superfamily of calcium/phospholipid-binding proteins. It is highly tissue-specific, being expressed only in intestinal and kidney epithelial cells. Alternative splicing generates two isoforms, both of which bind to rafts. In view of the lack of structural information supporting the physiological role of this annexin subfamily, we have cloned, expressed and purified human annexin A13b to investigate its structural and functional properties. The N-terminus of annexin A13b: (i) destabilizes the conserved protein core, as deduced from the low melting temperature in the absence (44 degrees C) or presence of calcium (55 degrees C), and (ii) impairs calcium dependent binding to acidic phospholipids, requiring calcium concentrations >400 microM. Truncation of the N-terminus restores thermal stability and decreases the calcium requirement for phospholipid binding, confirming its essential role in the structure-function relationship of this annexin. Non-myristoylated annexin A13b only binds to acidic phospholipids at high calcium concentrations. We show for the first time that myristoylation of annexin A13b enables the direct binding to phosphatidylcholine, raft-like liposomes and acidic phospholipids in a calcium independent manner. The conformational switch induced by calcium binding, from a 'closed' to an 'open' conformation with exposure of Trp227, can be mimicked by a decrease in pH, a process that may be relevant for membrane interactions. Our studies confirm that the common structural and functional characteristics that are dependent on the protein core of vertebrate annexins are likely to be common conserved features, whereas their variable N-termini confer distinct functional properties on annexins, as we report for myristoylation of annexin A13b. PMID- 15813709 TI - Modernizing UK health services: 'short-sharp-shock' reform, the NHS subsistence economy, and the spectre of health care famine. AB - Modernization is the trend for societies to grow functionally more complex, efficient and productive. Modernization usually occurs by increased specialization of function (e.g. division of labour, such as the proliferation of specialists in medicine), combined with increased organization in order to co ordinate the numerous specialized functions (e.g. the increased size of hospitals and specialist teams, including the management of these large groups). There have been many attempts to modernize the National Health Service (NHS) over recent decades, but it seems that none have significantly enhanced either the efficiency or output of the health care system. The reason may be that reforms have been applied as a 'drip-drip' of central regulation, with the consequence that health care has become increasingly dominated by the political system. In contrast, a 'short-sharp-shock' of radical and rapid modernization seems to be a more successful strategy for reforming social systems - in-between waves of structural change the system is left to re-orientate towards its client group. An example was the Flexner-initiated reform of US medical education which resulted in the closure of nearly half the medical colleges, an immediate enhancement in quality and efficiency of the system and future growth based on best institutional practices. However, short-sharp-shock reforms would probably initiate an NHS 'health care famine' with acute shortages and a health care crisis, because the NHS constitutes a 'subsistence economy' without any significant surplus of health services. The UK health care system must grow to generate a surplus before it can adequately be modernized. Efficient and rapid growth in health services could most easily be generated by stimulating provision outside the NHS, using mainly staff trained abroad and needs-subsidized 'item-of-service'-type payment schemes. Once there is a surplus of critically vital health services (e.g. acute and emergency provision), then radical modernization should rapidly improve the health service by a cull of low-quality and inefficient health care providers. PMID- 15813708 TI - Scientific evaluation of community-based Parkinson's disease nurse specialists on patient outcomes and health care costs. AB - OBJECTIVE: To determine the effects of community based nurses specialising in Parkinson's disease on health outcomes and healthcare costs. DESIGN: Two year randomised controlled trial in 438 general practices in nine randomly selected health authority areas of England. PARTICIPANTS: 1859 patients with Parkinson's disease identified by the participating general practices. MAIN OUTCOME MEASURES: Survival, stand-up test, dot in square test, bone fracture, global health question, PDQ-39, Euroqol, and healthcare costs. RESULTS: After two years 315 (17.3%) patients had died, although mortality did not differ between those who were attended by nurse specialists and those receiving standard care from their general practitioner (hazard ratio for nurse group v control group 0.91, 95% confidence interval 0.73 to 1.13). No significant differences were found between the two groups for the stand-up test (odds ratio 1.15, 0.93 to 1.42) and dot in square score (difference -0.7, -3.25 to 1.84). Scores on the global health question were significantly better in patients attended by nurse specialists than in controls (difference -0.23, -0.4 to -0.06), but no difference was observed in the results of the PDQ-39 or Euroqol questionnaires. Direct costs for patient health care increased by an average of pound2658 during the study, although not differentially between groups: the average increase was pound266 lower among patients attended by a nurse specialist (- pound981 to pound449). CONCLUSIONS: Nurse specialists in Parkinson's disease had little effect on the clinical condition of patients, but they did improve their patients' sense of wellbeing, with no increase in patients' healthcare costs. PMID- 15813710 TI - Evaluation of drug therapy and risk factors in diabetic hypertensives: a study of the quality of care provided in diabetic clinics in Bahrain. AB - OBJECTIVE: To evaluate control of blood pressure (BP) and diabetes and the associated risk factors in diabetic hypertensives treated by diabetic clinic primary care physicians. METHODS: A retrospective analysis of the medical records of diabetic hypertensives from six primary care diabetic clinics in Bahrain. RESULTS: The recommended BP target <130/<85 mmHg and of glycosylated haemoglobin (HbA1C) <7% were achieved in 7.5% and 14.5%, respectively. Most of the patients with uncontrolled BP and HbA(1C) were at high cardiovascular risk. More patients were on antihypertensive monotherapy than on combination therapy (60.6% vs. 36.7%; P<0.0001). The recommended two- and three-antihypertensive drug combinations were less often prescribed. In high-risk patients glycaemic control achieved was poor: antidiabetic combination therapy vs. monotherapy did not significantly differ. Inappropriate prescribing practices, such as the use of immediate-release nifedipine monotherapy, use of sulphonylurea instead of metformin in obese patients, and a trend towards prescribing of glyburide rather than a gliclazide in the elderly, were observed. Lipid-lowering (13.5%) and antiplatelet (12.8%) drugs were infrequently prescribed. CONCLUSIONS: Hypertension and diabetes in patients treated at the primary care diabetic clinics were inadequately controlled. In several instances, mono- and combination antihypertensives prescribed were irrational. Lipid-lowering and platelet aggregation inhibition strategies have received little attention. Intensive antihypertensive and antidiabetic complementary combination therapy should be encouraged. Continuous professional education of diabetic clinic physicians and expert-supervised diabetic clinics are desirable. PMID- 15813711 TI - Use of a patient information system to audit the introduction of modified early warning scoring. AB - Modified early warning scoring (MEWS) uses abnormalities in routine observations to identify patients at risk of critical illness. Nurses recorded scores at or above the medical response score of 3 on a hospital clinical information system during the first year of introducing MEWS to 10 wards in a university hospital. A total of 619 triggers were recorded in 365 patients. Fifty-nine required intensive care unit (ICU)/high dependency unit (HDU) care; 71 died. Survival was significantly worse for initial scores >4 (35/104 patients died) than for scores 3-4 (P<0.004). Multivariant analysis showed age (P<0.001) and trigger score (P<0.001) but not ward specialty (P=0.1) predicted death. Mean ages of survivors and non-survivors were 64 years (SD 18) and 74 years (SD 17), respectively. Addition of a score for age did not significantly increase the area under a receiver operator characteristic curve for the predictive value of MEWS scores. The study shows that increasing MEWS score is associated with worse outcome across a range of specialties and that nursing staff will use a patient information system to audit MEWS scores. PMID- 15813712 TI - Computer-aided disease prediction system: development of application software with SAS component language. AB - AIMS: The intricacy of predictive models associated with prognosis and risk classification of disease often discourages medical personnel who are interested in this field. The aim of this study was therefore to develop a computer-aided disease prediction model underpinning a step-by-step statistics-guided approach including five components: (1) data management; (2) exploratory analysis; (3) type of predictive model; (4) model verification; (5) interactive mode of disease prediction using SAS 8.02 Windows 2000 as a platform. METHODS: The application of this system was illustrated by using data from the Swedish Two-County Trial on breast cancer screening. The effects of tumour size, node status, and histological grade on breast cancer death using logistic regression model or survival models were predicted. A total of 20 questions were designed to exemplify the usefulness of each component. We also evaluated the system using a controlled randomized trial. Times to finish the above 20 questions were used as endpoint to evaluate the performance of the current system. User satisfaction with the current system such as easy to use, the efficiency of risk prediction, and the reduction of barrier to predictive model was also evaluated. RESULTS: The intervention group not only performed more efficiently than the control group but also satisfied with this application software. CONCLUSIONS: The MD-DP-SOS system characterized by menu-driven style, comprehensiveness, accuracy and adequacy assessment, and interactive mode of disease prediction is helpful for medical personnel who are involved in disease prediction. PMID- 15813713 TI - Understanding safer practices in health care: a prologue for the role of indicators. AB - OBJECTIVES: Patient safety and safer practices are central themes to many national strategies for accountability. The multinational Quality Indicator Project (QI Project) database is used to identify patterns of indicator use to measure safety of care in Asia, Europe, and the USA. The second objective is to assess, within the context of an indicator project, the usefulness of indicators to measure errors or mishaps. DESIGN AND SETTING: This descriptive study retrospectively analyses indicator use patterns among hospitals in Asia, Europe, and the USA. The QI Project database is used for the 1999-2002 period. Statistical testing (P-value) of the differences in use percentages across five countries is based on 'country' rather than 'hospital' as the unit of analysis. RESULTS: There was a significant increase in overall QI Project indicator use worldwide between 1999 and 2002. The average change in use was 6.8% for safety indicators and 4.2% for all other indicators. When analysed by country (USA, Austria, Belgium, UK, and Taiwan), the average increases in use percentage were highest in Taiwan and Belgium. When the country-specific differences were tested for significance, Taiwan showed the largest (and statistically significant) increase in safety indicator use between 1999 and 2002 (P<0.0001). In the USA, the rates of safety indicator use have decreased (P=0.0502) during the same time period. CONCLUSION: This paper identifies, perhaps for the first time, how traditional indicators of hospital performance are being used to understand a hospital's performance and associated safety of care. Although the study's time frame is limited to 3 years, the findings seem to suggest that the interest in using these traditional indicators as proxies for safer practice measures is increasing among the QI Project participants worldwide. The challenge of using inherently value-free indicators as indicators of safety (hence de facto labelled as 'error' focused) should be further studied. PMID- 15813714 TI - Medicine and literature: writing and reading. AB - The humanities and arts are appropriate areas of study within interdisciplinary medicine. Medicine has long been considered to be both a science and an art. Within each patient, the psychological, emotional, spiritual, and the physical are all inextricably linked. The values, ideas and images of individuals and culture, as well as the way the human body and mind physically function and dysfunction, impinge in an equally inextricable way upon all these elements. The humanities and arts, included within medical study, enable people, the subject of medicine, to be usefully considered in their entirety. Literature has a particularly vital role to play in medicine and health care. It opens up a wealth of experience and knowledge, as well as offering vital understandings of the narrative nature of human lives. A knowledge of the nature of narrative, and the way we understand our lives narratively can be vital for effective communication and understanding of patients' situations. Literature also offers dynamic ethical issues with which to grapple. Expressive and explorative writing is used in professional development, and undergraduate courses. Medical humanities, literature and medicine and narrative medicine are established in the USA and developing fast in Britain. PMID- 15813715 TI - Predictive survival model with time-dependent prognostic factors: development of computer-aided SAS Macro program. AB - AIMS AND OBJECTIVES: Computer program for the prediction of survival with respect to time-dependent proportional hazards regression model has been rarely addressed. We therefore developed a SAS Macro program for time-dependent Cox regression predictive model for empirical survival data associated with time dependent covariates. METHOD: Time-dependent proportional hazards regression model and partial likelihood in association with time-varying predictors were explicitly delineated. Baseline hazard using Andersen's method was incorporated into proportional hazards regression model to predict the dynamic change of cumulative survival in respect of time-varying predictors. Two SAS Macro programs for time-dependent predictive survival model and model validation using receiver operative characteristics were written with SAS IML language. RESULTS: The computer program was applied to data on clinical surveillance of small hepatocellular carcinoma (HCC) treated by percutaneous ethanol injection (PEI) or transcatheter arterial embolization (TAE) with time-varying predictors such as alpha-feto protein (AFP) and other biological markers. CONCLUSION: The program is very useful for real-time prediction of cumulative survival on the basis of time dependent covariates. PMID- 15813716 TI - Unity from diversity: the evidential use of anecdotal reports of adverse drug reactions and interactions. AB - Anecdotal case reports contribute about one-third of the published literature on adverse drug reactions and interactions, but are regarded as providing poor quality evidence. However, they can occasionally provide proof of cause and effect, and there are many other reasons for publishing them. Because an anecdote is a narrative, narratological paradigms from literature, art, and music can show how we can make evidential use of anecdotes. Useful paradigms are the dramatic unities (of time, place, and action), comprehensive catalogues, and pattern formations. Here I give examples of each of these types of paradigm and show how they can be used to interpret anecdotes about adverse drug reactions and interactions. The dramatic unities show how a proper classification of adverse drug reactions can be achieved, according to dose-relation, time-course, and susceptibility factors; use of this classification should improve the evidential use of anecdotal reports. A high background incidence of the effect (the medical equivalent of subplots, which violate the unity of action) makes it more difficult to detect adverse drug effects using anecdotal reports. To make best evidential use of the corpus of anecdotal reports of adverse drug reactions, comprehensiveness is important: each suspected adverse reaction should be reported in detail and reactions should be reported in sufficient numbers for proper classification and for patterns to be recognized. One form of pattern recognition, teleoanalysis of data, should, when possible, include not only randomized controlled trials and observational studies, but also case series and anecdotal reports. PMID- 15813717 TI - The pathogenicity of Entamoeba histolytica is related to the capacity of evading innate immunity. AB - The host and parasite factors that influence susceptibility to Entamoeba histolytica infection and disease are not well understood. Entamoeba histolytica pathogenicity has been considered by focusing principally on parasite rather than host factors. Thus, research has concentrated on explaining the molecular differences between pathogenic E. histolytica and non-pathogenic E. dispar. However, the amoeba molecules considered most important for host tissue destruction (amoebapore, galactose/N-acetyl galactosamine inhibitable lectin, and cysteine proteinases) are present in both pathogenic E. histolytica and non pathogenic E. dispar. In addition, the genetic differences in pathogenicity among E. histolytica isolates are unlikely to completely explain the different outcomes of infection. Considering that the principal difference between pathogenic and non-pathogenic amoebas lies in their surface coats, we propose that pathogenicity of the amoebas is related to the composition and properties of the surface coat components (or pathogen-associated molecular patterns, PAMPs), and the ability of innate immune response to recognize these components and eliminate the parasite. According to this hypothesis, a key feature that may distinguish pathogenic (E. histolytica) from non-pathogenic (E. dispar) strains is whether or not they can overcome innate immune defences. A corollary of this hypothesis is that in susceptible individuals the PAMPs are either not recognized or they are recognized by a set of Toll-like receptors (TLRs) that leads to an inflammatory response. In both cases, the result is tissue damage. On the contrary, in resistant individuals the innate/inflammatory response, induced through the activation of a different set of TLRs, eliminates the parasite. PMID- 15813718 TI - Soluble cellular adhesion molecules, selectins, VEGF and endothelin-1 in patients with Wuchereria bancrofti infection and association with clinical status. AB - Lymphatic filariasis, a mosquito-transmitted disease commonly known as Bancroftian filariasis, is characterized by debilitating pathology linked to the progression of lymphoedema to a chronic state of elephantiasis. We performed longitudinal measurements of endothelial adhesion and angiogenic molecules in 63 Polynesian patients living in an hyperendemic focus of Wuchereria bancrofti. Decreased serum concentrations of soluble (s-) L selectin (CD62L) were noticed in sera of of patients with chronic conditions (hydrocele and elephantiasis). Chyluria was associated with increased vascular endothelial growth factor (VEGF) levels, whereas elephantiasis presented a high endothelin-1 (ET-1) profile. By contrast, increased serum concentrations of soluble intercellular (sICAM-1, CD54), but not of vascular cell (sVCAM-1, CD106), adhesion molecules were observed in sera of patients with bacterial lymphangitis used as controls. These trends are consistent with the increased permeability of vascular structures, a major clinical feature observed in acute lymphatic pathology (of bacterial or filarial origin), and of fundamental differences in the pathogenesis of hydrocele and elephantiasis. Using markers correlated with the clinical status (high ET-1 and VEGF levels for elephantiasis and chyluria, respectively; low CD62L levels for hydrocoele and elephantiasis) it should be possible to monitor disease progression in lymphatic filariasis. PMID- 15813719 TI - Mucosal cytokine and antigen-specific responses to Cryptosporidium parvum in IL 12p40 KO mice. AB - Studies of cellular immune responses to Cryptosporidium parvum have been limited in part by lack of suitable animal models. IL-12p40(-/-)mice are susceptible to initial infection with C. parvum but recover within 2 weeks, rendering the animals resistant to reinfection. Because the host responses that determine duration and severity of primary infection are not yet understood, we studied the cellular immune response to primary infection with C. parvum in IL-12p40(-/-)mice and also explored possible mechanisms for this response. Female IL-12p40(-/-)mice were inoculated with 10,000 oocysts. Uninfected age-matched mice served as controls. At different time intervals following exposure to oocysts, mice were sacrificed and their intestine, spleen, and mesenteric lymph node tissues were harvested. Cellular immune responses to C. parvum were characterized. Infection of IL-12p40(-/-)mice induced changes in the gene expression of the cytokines IFN gamma, IL-4, IL-15, IL-18, TNF-alpha and TGF-beta during primary infection. There was also a significant increase in total numbers of lymphocytes and CD19/CD62L expressing cells in mesenteric lymph nodes. These MLN cells exhibited increased antigen-specific proliferation and cytokine production (IL-6 and IFN-gamma) levels when stimulated in vitro. These observations delineate the cellular immune responses during acute C. parvum infection of the IL-12p40(-/-)mouse model. PMID- 15813721 TI - Cytokine mRNA expressions in symptomatic vs. asymptomatic amoebiasis patients. AB - Infection with Entamoeba histolytica results in high mortality worldwide. Studies on the cytokine response in symptomatic and asymptomatic amoebiasis (caused by E. histolytica, the pathogenic species and E. dispar, the non-pathogenic species) subjects and their correlation with symptomatology are lacking. The present study reports the cytokine response (TNF-alpha, IFN-gamma, IL-4, IL-10 and TGF-beta) in such subjects as measured by RT-PCR. The results showed significantly (< 0.05) higher expressions of IL-10 and TGF-beta in the symptomatic group as compared to the asymptomatic and healthy controls. The cytokine profile indicated the role of suppressive immune response in symptomatic amoebiasis patients. PMID- 15813720 TI - Oral immunization with a live recombinant attenuated Salmonella typhimurium protects mice against Toxoplasma gondii. AB - The natural site of infection for T. gondii is the mucosal surface of the intestine, so the protective immunity obtained after natural infection with T. gondii points to the importance of developing a vaccine that stimulates mucosal defences. In this study, an aroA- and aroD- attenuated strain of Salmonella typhimurium (BRD509) has been used to deliver the recombinant eukaryotic plasmid pSAG(1-2)/CTA2/B expressing a multi-antigenic gene encoding SAG1 and SAG2 of T. gondii linked to A2/B subunits of cholera toxin as a candidate oral T. gondii vaccine. Immunoblot analysis showed compound gene expression in HeLa cells in vitro and intragastric immunization of mice with the recombinant salmonella resulted in the induction of humoral and Th1 type cellular immune responses and afforded protection against RH strain T. gondii challenge. Anti-T. gondii IgG values increased markedly in the BRD509/pSAG(1-2)-CTA2/B immunized group; these values were significantly higher than those in the negative controls (P = 0.008). With CTA2/B genetic adjuvant, the T. gondii-specific response was predominantly Th1, indicating that the CTA(2)/B genetic adjuvant was able to overcome the strong Th2-bias of the antigen (IgG2a >> IgG1). Antigen-specific T cell proliferative responses and CTL activity were significantly enhanced when cholera toxin CTA2/B genetic adjuvant was used (P = 0.009; P = 0.006). Culture supernatants from antigen-stimulated splenocytes from mice in these groups were also examined by ELISA for Th1- and Th2-type cytokines; mean IFN-gamma levels produced after oral immunization with BRD509/pSAG(1-2)-CTA2/B were about nine fold higher than after immunization with BRD509/pSAG(1-2) (P = 0.007). On the other hand, the levels of IL-4 were low for all groups and no increase was seen in the presence of CTA2/B genetic adjuvant. When the immunized mice were intraperitoneally challenged with 10(3) tachyzoites of the highly virulent RH strain, the survival time of the mice immunized with BRD509/pSAG(1-2)-CTA2/B was markedly longer than other groups (P = 0.003) and a 40% survival rate was achieved. This is the first report that demonstrates that an oral attenuated salmonella DNA vaccine can induce protective immunity against the acute phase of T. gondii infection. PMID- 15813722 TI - PD-L2+ dendritic cells and PD-1+ CD4+ T cells in schistosomiasis correlate with morbidity. AB - Dendritic cells (DC) are critical antigen-presenting cells for the induction and control of immune responses. PD-L2 (B7-DC) is a regulatory ligand on subpopulations of DC, and binds to the co-regulatory receptor PD-1, present on some activated T lymphocytes, leading to down-regulation. We now show that very early during experimental schistosomiasis (by 5 weeks) a significantly higher proportion of splenic CD11c+/B220- DC express PD-L2, and by 6 weeks after infection a higher proportion of splenic CD4 T cells express PD-1. In this CBA/J mouse/Schistosoma mansoni chronic infection model we have shown that most mice develop moderate morbidity (Moderate Splenomegaly Syndrome, MSS), while some parallel-infected mice express different immune characteristics and die or develop severe morbidity (Hypersplenomegaly Syndrome, HSS). We now report a positive correlation between the proportion of splenic CD11c+/B220- DC that express PD-L2 and showing MSS. In contrast, there is an inverse correlation between the proportion of splenic CD3+/CD4+ T lymphocytes that express PD-1 and showing MSS. The data demonstrate that schistosomes can induce sustained elevated percentages of PD-L2-expressing, B220-negative DC. Furthermore, when this potentially immunoregulatory environment occurs chronically, infected mice are most likely to have developed MSS, expressing moderate morbidity. PMID- 15813723 TI - Presence of the LDNF glycan on the host-protective H-gal-GP fraction from Haemonchus contortus. AB - Immunization of sheep with the gut membrane-associated protein complex H-gal-GP of adult Haemonchus contortus induces high levels of protection against a homologous challenge infection. Protection is correlated with a systemic IgG response against the antigen. Analysis of the antibody response showed that the majority of the antigen-specific IgG was of the IgG2 isotype. A substantial proportion (74%) of this response was directed against the glycan component of H gal-GP. The high immunogenicity of the H-gal-GP glycans may be due to the presence of the fucosylated LacdiNAc (LDNF) antigen. 2D electrophoresis, Western blotting and mass spectrometry analysis of H-gal-GP showed that this glycan epitope was specifically located on a metalloendopeptidase, MEP3. MEP3 is the most abundant protein in H-gal-GP and has been identified as one of the most likely protective components of the complex. Here, we present evidence that the LDNF glycan does not contribute to the protective capacity of H-gal-GP. Animals vaccinated with reduced and denatured H-gal-GP are not protected against subsequent infection, although the antibody response against the LDNF glycan is very similar to that of animals vaccinated with the native H-gal-GP. In addition, an alternative version of H-gal-GP, H-sialgal-GP, which is equally protective, but isolated by affinity chromatography on jacalin lectin rather than peanut lectin, contains a MEP3 component which has no detectable LDNF glycan. PMID- 15813724 TI - A kinase for light and time. AB - The eukaryotic filamentous fungus Neurospora crassa has proven to be a dependable model system for the analysis of light-regulated gene expression and circadian rhythmicity. The molecular basis of the core circadian clock is a transcription/translation-based feedback loop in which a heterodimer of the white collar 1 (WC-1) and white-collar 2 (WC-2) proteins act as a transcription factor to promote expression of the negative elements in the clock, frq mRNA and FRQ protein. Additionally, WC-1 is a flavoprotein that acts as the primary responder to environmental light and, in conjunction with WC-2, is the major photoreceptor for the clock as well as other light-regulated processes. Protein kinase C acts as a light-dependent regulator of the WC-1 protein. PMID- 15813725 TI - Measure for measure in the control of type III secretion hook and needle length. AB - Bacterial flagella and injectisomes are supramolecular structures that are responsible for motility and for delivering toxic proteins into the cytosol of eukaryotic cells, respectively. They look very similar to each other. Both systems are called type III secretion pathways, and their components share substantial sequence similarities. One remarkable feature of the type III systems is that the length of their substructure is fairly well controlled by a secretion switching machinery, which consists of at least two proteins, a length control protein and an integral membrane secretion component. Here, we review how and why the length of these structures must be accurately controlled. PMID- 15813726 TI - Understanding the control of Pseudomonas aeruginosa alginate synthesis and the prospects for management of chronic infections in cystic fibrosis. AB - Decades of research have been dedicated to the study of the opportunistic pathogen Pseudomonas aeruginosa, a Gram-negative, environmental bacterium that secretes the exopolysaccharide alginate during chronic lung infection of cystic fibrosis (CF) patients. Although P. aeruginosa utilizes a variety of factors to establish a successful infection in the lungs of CF patients, alginate has stood out as one of the best-studied prognostic indicators of chronic lung infection. While the genetics, biosynthesis and regulation of alginate are well understood, questions still remain concerning its role in biofilm development and its potential as a therapeutic target. The purpose of this review is to provide a brief summary of alginate biosynthesis and regulation, and to highlight recent discoveries in the areas of alginate production, biofilm formation and vaccine design. This information is placed in context with a proposed P. aeruginosa infectious pathway, highlighting avenues for the use of existing therapies as well as the potential for novel agents to reduce or eliminate chronic infections in CF patients. PMID- 15813727 TI - Defining the role of PfCRT in Plasmodium falciparum chloroquine resistance. AB - Recent studies have highlighted the importance of a parasite protein referred to as the chloroquine resistance transporter (PfCRT) in the molecular basis of Plasmodium falciparum resistance to the quinoline antimalarials. PfCRT, an integral membrane protein with 10 predicted transmembrane domains, is a member of the drug/metabolite transporter superfamily and is located on the membrane of the intra-erythrocytic parasite's digestive vacuole. Specific polymorphisms in PfCRT are tightly correlated with chloroquine resistance. Transfection studies have now proven that pfcrt mutations confer verapamil-reversible chloroquine resistance in vitro and reveal their important role in resistance to quinine. Available evidence is consistent with the view that PfCRT functions as a transporter directly mediating the efflux of chloroquine from the digestive vacuole. PMID- 15813728 TI - Protein kinase C modulates light responses in Neurospora by regulating the blue light photoreceptor WC-1. AB - The Neurospora protein kinase C (NPKC) is a regulator of light responsive genes. We have studied the function of NPKC in light response by investigating its biochemical and functional interaction with the blue light photoreceptor white collar 1 (WC-1), showing that activation of NPKC leads to a significant decrease in WC-1 protein levels. Furthermore, we show that WC-1 and NPKC interact in a light-regulated manner in vivo, and that protein kinase C (PKC) phosphorylates WC 1 in vitro. We designed dominant negative and constitutively active forms of PKC which are able to induce either a large increase of WC-1 protein level or a strong reduction respectively. Moreover, these changes in PKC activity result in an altered light response. As WC-1 is a key component of Neurospora circadian clock and regulates the clock oscillator component FRQ we investigated the effect of NPKC-mutated forms on FRQ levels. We show that changes in PKC activity affect FRQ levels and the robustness of the circadian clock. Together these data identify NPKC as a novel component of the Neurospora light signal transduction pathway that modulates the circadian clock. PMID- 15813729 TI - N-terminal signal region of FliK is dispensable for length control of the flagellar hook. AB - The length of the flagellar hook is regulated; it is 55 +/- 6 nm long in Salmonella. Five genes involved in hook-length regulation are fliK, flhB, fliG, fliM and fliN. The last four genes encode structural components of the protein export apparatus in the flagellar base, whereas FliK is soluble and secreted during flagellar assembly. The role of FliK, however, remains ambiguous. We constructed two kinds of FliK variants: N-terminally truncated FliK protein and FliK N-terminally fused with cyan fluorescent protein (CFP-FliK). Both N terminally truncated FliK missing the first 99 amino acids (aa) and CFP-FliK fusion variants partially complemented a fliK null (polyhook) mutant to produce cells with filaments, allowing cells to swim; the hooks, however, were not normal but were polyhooks. When the N-terminally defective FliK variants were expressed at high levels, the average polyhook length was shortened coming close to the length of the wild-type hook, independently of the sizes of the FliK variants. These FliK variants were not secreted. CFP-FliK fusion proteins were observed to homogeneously distribute in the cytoplasm. We conclude that FliK does not need to be exported to control hook length and is unlikely to be a ruler; instead, we conclude that FliK controls hook length by the timely switching of secretion modes of the flagellar type III secretion system by the FliK C-terminal domain, and that the N-terminal region is dispensable for hook length control. PMID- 15813730 TI - The unique structure of archaeal 'hami', highly complex cell appendages with nano grappling hooks. AB - Proteinaceous, hair-like appendages known as fimbriae or pili commonly extend from the surface of prokaryotic cells and serve important functions such as cell adhesion, biofilm formation, motility and DNA transfer. Here we show that a novel group of archaea from cold, sulphidic springs has developed cell surface appendages of an unexpectedly high complexity with a well-defined base-to-top organization. It represents a new class of filamentous cell appendages, for which the term 'hamus' is proposed. Each archaeal cell is surrounded by a halo of about 100 hami, which mediate strong adhesion of the cells to surfaces of different chemical composition. The hami are mainly composed of 120 kDa subunits and remained stable in a broad temperature and pH range (0-70 degrees C; 0.5-11.5). Electron microscopy and cryo-electron tomography revealed that the hamus filament possesses a helical basic structure. At periodic distances, three prickles emanate from the filament, giving it the character of industrially produced barbwire. At its distal end the hami carry a tripartite, barbed grappling hook (60 nm in diameter). The architecture of this molecular hook is reminiscent of man-made fishhooks, grapples and anchors. It appears that nature has developed a perfect mechanical nano-tool in the course of biological evolution, which also might prove useful in the field of nanobiotechnology. PMID- 15813731 TI - Regulation of Sin recombinase by accessory proteins. AB - Sin recombinase from Staphylococcus aureus acts selectively on directly repeated resH sites, assembling an intertwined synapse in which exactly three supercoils are trapped between the points of strand exchange. Resolution requires the two Sin binding sites in resH (site I, where strand exchange occurs, and site II) and a non-specific DNA-bending protein (e.g. Hbsu). We show that a single amino acid substitution in Sin (I100T) is sufficient to relax the normal requirements for site II and Hbsu. Using this hyperactive protein, and the variant recombination site resH(AT), we investigate the roles of site II and Hbsu in synapsis and strand exchange. We conclude that Sin bound at site II, and Hbsu, act together to control site I alignment and the topology of the synapse, and to stimulate strand exchange. PMID- 15813732 TI - Translation elongation factor EF-Tu is a target for Stp, a serine-threonine phosphatase involved in virulence of Listeria monocytogenes. AB - Listeria monocytogenes is a pathogen that causes listeriosis, a severe food-borne infection. This bacterium, in order to survive and grow in the multiple conditions encountered in the host and the environment, has evolved a large number of regulatory elements, in particular many signal transduction systems based on reversible phosphorylation. The genome sequence has revealed genes for 16 putative two-component systems, four putative tyrosine phosphatases, three putative serine-threonine kinases and two putative serine-threonine phosphatases. We found that one of the latter genes, stp, encodes a functional Mn(2+)-dependent serine-threonine phosphatase similar to PPM eukaryotic phosphatases (Mg(2+)-or Mn(2+)-dependent protein phosphatase) and is required for growth of L. monocytogenes in a murine model of infection. We identified as the first target for Stp, the elongation factor EF-Tu. Post-translational phosphorylation of EF-Tu had been shown to prevent its binding to amino-acylated transfer RNA as well as to kirromycin, an antibiotic known to inhibit EF-Tu function. Accordingly, an stp deletion mutant is less sensitive to kirromycin. These results suggest an important role for Stp in regulating EF-Tu and controlling bacterial survival in the infected host. PMID- 15813733 TI - Granulocytes govern the transcriptional response, morphology and proliferation of Candida albicans in human blood. AB - Survival in blood and escape from blood vessels into tissues are essential steps for the yeast Candida albicans to cause systemic infections. To elucidate the influence of blood components on fungal growth, morphology and transcript profile during bloodstream infections, we exposed C. albicans to blood, blood fractions enriched in erythrocytes, polymorphonuclear or mononuclear leukocytes, blood depleted of neutrophils and plasma. C. albicans cells exposed to erythrocytes, mononuclear cells, plasma or blood lacking neutrophils were physiologically active and rapidly switched to filamentous growth. In contrast, the presence of neutrophils arrested C. albicans growth, enhanced the fungal response to overcome nitrogen and carbohydrate starvation, and induced the expression of a large number of genes involved in the oxidative stress response. In particular, SOD5, encoding a glycosylphosphatidylinositol (GPI)-anchored superoxide dismutase localized on the cell surface of C. albicans, was strongly expressed in yeast cells that were associated with neutrophils. Mutants lacking key genes involved in oxidative stress, morphology or virulence had significantly reduced survival rates in blood and the neutrophil fraction, but remained viable for at least 1 h of incubation when exposed to erythrocytes, mononuclear cells, plasma or blood lacking neutrophils. These data suggest that C. albicans genes expressed in blood were predominantly induced in response to neutrophils, and that neutrophils play a key role during C. albicans bloodstream infections. However, C. albicans is equipped with several genes and transcriptional programmes, which may help the fungus to counteract the attack of neutrophils, to escape from the bloodstream and to cause systemic infections. PMID- 15813734 TI - Nck-independent actin assembly is mediated by two phosphorylated tyrosines within enteropathogenic Escherichia coli Tir. AB - Enteropathogenic Escherichia coli (EPEC) stimulates tyrosine-kinase signalling cascades to trigger localized actin assembly within mammalian cells. During actin 'pedestal' formation, the EPEC effector protein Tir is translocated into the plasma membrane, becomes phosphorylated on tyrosine-474 (Y474) and promotes recruitment of the mammalian adaptor protein Nck to efficiently activate N-WASP Arp2/3-mediated actin polymerization. Tir also triggers localized actin assembly in the absence of Nck, but the Tir sequences involved in this signalling cascade have not been defined. To identify and characterize the phosphotyrosines that contribute to Nck-independent pedestal formation, we investigated the regulation of Tir tyrosine phosphorylation and found that phosphorylation is stimulated by Tir clustering. In addition to Y474, residue Y454 is also phosphorylated, although at lower efficiency. These tyrosines differentially contribute to actin polymerization in a fashion reminiscent of actin 'tail' formation mediated by the vaccinia virus envelope protein A36R, which utilizes two similarly spaced phosphotyrosines to recruit the adaptors Nck and Grb2, respectively, in order to stimulate N-WASP. Neither phosphorylated Y454 nor Y474 directly bind Grb2, but Tir derivatives harbouring these residues ultimately recruit N-WASP and Arp2/3 independently of Nck, suggesting that EPEC exploits additional phosphotyrosine binding adaptors capable of initiating actin assembly. PMID- 15813735 TI - The mutation G145S in PrfA, a key virulence regulator of Listeria monocytogenes, increases DNA-binding affinity by stabilizing the HTH motif. AB - Listeria monocytogenes, a Gram-positive, facultative intracellular human pathogen, causes systemic infections with high mortality rate. The majority of the known pathogenicity factors of L. monocytogenes is regulated by a single transcription factor, PrfA. Hyperhaemolytic laboratory strains of L. monocytogenes express the constitutively active mutant PrfA(G145S) inducing virulence gene overexpression independent of environmental conditions. PrfA belongs to the Crp/Fnr family of transcription factors generally activated by a small effector, such as cAMP or O(2). We present the crystal structures of wild type PrfA, the first Gram-positive member of the Crp/Fnr family, and of the constitutively active mutant PrfA(G145S). Cap (Crp) has previously been described exclusively in the cAMP-induced (DNA-free and -bound) conformation. By contrast, the PrfA structures present views both of the non-induced state and of the mutationally activated form. The low DNA-binding affinity of wild-type PrfA is supported both structurally (partly disordered helix-turn-helix motif, overall geometry of the HTH alpha-helices deviates from Cap) and by surface plasmon resonance analyses (K(D) = 0.9 microM). In PrfA(G145S) the HTH motifs dramatically rearrange to adopt a conformation comparable to cAMP-induced Cap and hence favourable for DNA binding, supported by a DNA-binding affinity of 50 nM. Finally, the hypothesis that wild-type PrfA, like other Crp/Fnr family members, may require an as yet unidentified cofactor for activation is supported by the presence of a distinct tunnel in PrfA, located at the interface of the beta barrel and the DNA-binding domain. PMID- 15813736 TI - Enteropathogenic Escherichia coli EspG disrupts microtubules and in conjunction with Orf3 enhances perturbation of the tight junction barrier. AB - EspG, a secreted effector of enteropathogenic Escherichia coli (EPEC), as well as its homologue Orf3, has been shown to disrupt microtubules (MTs) in fibroblasts and non-polarized epithelial cells. The roles of MTs and the effects of MT disruption in these cell types differ significantly. The aim of this study was to investigate the effects of EspG on polarized, host target intestinal epithelial cells. Immunofluorescent labelling of tubulin showed that EPEC caused progressive fragmentation and loss of the MT network in cells harbouring attached organisms. Immunoblots of proteins extracted from EPEC-infected cells showed a corresponding loss of alpha-tubulin. Type III secretion system (TTSS)-deficient strains had no effect on MT suggesting TTSS dependence. Mutation of espG, but not espF or map, ablated EPEC's effects on MTs for up to 6 h. Ectopic expression of EspG in HeLa cells caused MT disruption. While deletion of espG alone had no effect on the EPEC-induced decrease in transepithelial electrical resistance (TER), mutation of both espG and orf3 significantly delayed the kinetics of this response. Complementation of the double mutant with espG alone restored the kinetics of TER drop to that of wild type. Herein, we describe a previously unrecognized phenotype for the EPEC effectors EspG and Orf3. PMID- 15813737 TI - A bacterial hormone (the SCB1) directly controls the expression of a pathway specific regulatory gene in the cryptic type I polyketide biosynthetic gene cluster of Streptomyces coelicolor. AB - Gamma-butyrolactone signalling molecules are produced by many Streptomyces species, and several have been shown to regulate antibiotic production. In Streptomyces coelicolor A3(2) at least one gamma-butyrolactone (SCB1) has been shown to stimulate antibiotic production, and genes encoding proteins that are involved in its synthesis (scbA) and binding (scbR) have been characterized. Expression of these genes is autoregulated by a complex mechanism involving the gamma-butyrolactone. In this study, additional genes influenced by ScbR were identified by DNA microarray analysis, and included a cryptic cluster of genes for a hypothetical type I polyketide. Further analysis of this gene cluster revealed that the pathway-specific regulatory gene, kasO, is a direct target for regulation by ScbR. Gel retardation and DNase I footprinting analyses identified two potential binding sites for ScbR, one at -3 to -35 nt and the other at -222 to -244 nt upstream of the kasO transcriptional start site. Addition of SCB1 eliminated the DNA binding activity of ScbR at both sites. The expression of kasO was growth phase regulated in the parent (maximal during transition phase), undetectable in a scbA null mutant, and constitutively expressed in a scbR null mutant. Addition of SCB1 to the scbA mutant restored the expression of kasO, indicating that ScbR represses kasO until transition phase, when presumably SCB1 accumulates in sufficient quantity to relieve kasO repression. Expression of the cryptic antibiotic gene cluster was undetectable in a kasO deletion mutant. This is the first report with comprehensive in vivo and in vitro data to show that a gamma-butyrolactone-binding protein directly regulates a secondary metabolite pathway-specific regulatory gene in Streptomyces. PMID- 15813738 TI - Blue light negatively regulates the sexual filamentation via the Cwc1 and Cwc2 proteins in Cryptococcus neoformans. AB - Cryptococcus neoformans is a heterothallic basidiomycetous yeast that primarily infects immunocompromised individuals. Dikaryotic hyphae resulting from the fusion of the MATa and MATalpha mating type strains represent the filamentous stage in the sexual life cycle of C. neoformans. In this study we demonstrate that the production of dikaryotic filaments is inhibited by blue light. To study blue light photoresponse in C. neoformans, we have identified and characterized two genes, CWC1 and CWC2, which are homologous to Neurospora crassa wc-1 and wc-2 genes. Conserved domain analyses indicate that the functions of Cwc1 and Cwc2 proteins may be evolutionally conserved. To dissect their roles in the light response, the CWC1 gene deletion mutants are created in both mating type strains. Mating filamentation in the bilateral cross of cwc1 MATa and MATalpha strains is not sensitive to light. The results indicate that Cwc1 may be an essential regulator of light responses in C. neoformans. Furthermore, overexpression of the CWC1 or CWC2 gene requires light activation to inhibit sexual filamentation, suggesting both genes may function together in the early step of blue light signalling. Taken together, our findings illustrate blue light negatively regulates the sexual filamentation via the Cwc1 and Cwc2 proteins in C. neoformans. PMID- 15813739 TI - Co-regulation of Salmonella enterica genes required for virulence and resistance to antimicrobial peptides by SlyA and PhoP/PhoQ. AB - Analysis of the transcriptome of slyA mutant Salmonella enterica serovar Typhimurium revealed that many SlyA-dependent genes, including pagC, pagD, ugtL, mig-14, virK, phoN, pgtE, pipB2, sopD2, pagJ and pagK, are also controlled by the PhoP/PhoQ regulatory system. Many SlyA- and PhoP/PhoQ-co-regulated genes have functions associated with the bacterial envelope, and some have been directly implicated in virulence and resistance to antimicrobial peptides. Purified His tagged SlyA binds to the pagC and mig-14 promoters in regions homologous to a previously proposed 'SlyA-box'. The pagC promoter lacks a consensus PhoP binding site and does not bind PhoP in vitro, suggesting that the effect of PhoP on pagC transcription is indirect. Stimulation of pagC expression by PhoP requires SlyA. Levels of SlyA protein and mRNA are not significantly changed under low-magnesium PhoP-inducing conditions in which pagC expression is profoundly elevated, however, indicating that the PhoP/PhoQ system does not activate pagC expression by altering SlyA protein concentration. Models are proposed in which PhoP may control SlyA activity via a soluble ligand or SlyA may function as an anti repressor to allow PhoP activation. The absence of almost all SlyA-activated genes from the Escherichia coli K12 genome suggests that the functional linkage between the SlyA and PhoP/PhoQ regulatory systems arose as Salmonella evolved its distinctive pathogenic lifestyle. PMID- 15813740 TI - Retrotransposition strategies of the Lactococcus lactis Ll.LtrB group II intron are dictated by host identity and cellular environment. AB - Group II introns are mobile retroelements that invade their cognate intron-minus gene in a process known as retrohoming. They can also retrotranspose to ectopic sites at low frequency. Previous studies of the Lactococcus lactis intron Ll.LtrB indicated that in its native host, as in Escherichia coli, retrohoming occurs by the intron RNA reverse splicing into double-stranded DNA (dsDNA) through an endonuclease-dependent pathway. However, in retrotransposition in L. lactis, the intron inserts predominantly into single-stranded DNA (ssDNA), in an endonuclease independent manner. This work describes the retrotransposition of the Ll.LtrB intron in E. coli, using a retrotransposition indicator gene previously employed in our L. lactis studies. Unlike in L. lactis, in E. coli, Ll.LtrB retrotransposed frequently into dsDNA, and the process was dependent on the endonuclease activity of the intron-encoded protein. Further, the endonuclease dependent insertions preferentially occurred around the origin and terminus of chromosomal DNA replication. Insertions in E. coli can also occur through an endonuclease-independent pathway, and, as in L. lactis, such events have a more random integration pattern. Together these findings show that Ll.LtrB can retrotranspose through at least two distinct mechanisms and that the host environment influences the choice of integration pathway. Additionally, growth conditions affect the insertion pattern. We propose a model in which DNA replication, compactness of the nucleoid and chromosomal localization influence target site preference. PMID- 15813741 TI - Respiration metabolism of Group B Streptococcus is activated by environmental haem and quinone and contributes to virulence. AB - Group B Streptococcus (GBS) is a common constituent of the vaginal microflora, but its transmission to newborns can cause life-threatening sepsis, pneumonia and meningitis. Energy metabolism of this opportunist pathogen has been deduced to be strictly fermentative. We discovered that GBS undergoes respiration metabolism if its environment supplies two essential respiratory components: quinone and haem. Respiration metabolism led to significant changes in growth characteristics, including a doubling of biomass and an altered metabolite profile under the tested conditions. The GBS respiratory chain is inactivated by: (i) withdrawing haem and/or quinone, (ii) treating cultures with a respiration inhibitor or (iii) inactivating the cydA gene product, a subunit of cytochrome bd quinol oxidase, in all cases resulting in exclusively fermentative growth. cydA inactivation reduced GBS growth in human blood and strongly attenuated virulence in a neonatal rat sepsis model, suggesting that the animal host may supply the components that activate GBS respiration. These results suggest a role of respiration metabolism in GBS dissemination. Our findings show that environmental factors can increase the flexibility of GBS metabolism by activating a newly identified respiration chain. The need for two environmental factors may explain why GBS respiration metabolism was not found in previous studies. PMID- 15813742 TI - Patterns of nucleosomal organization in the alc regulon of Aspergillus nidulans: roles of the AlcR transcriptional activator and the CreA global repressor. AB - We have studied the chromatin organization of three promoters of the alc regulon of Aspergillus nidulans. No positioned nucleosomes are seen in the aldA (aldehyde dehydrogenase) promoter under any physiological condition tested by us. In the alcA (alcohol dehydrogenase I) and alcR (coding for the pathway-specific transcription factor) promoters, a pattern of positioned nucleosomes is seen under non-induced and non-induced repressed conditions. While each of these promoters shows a specific pattern of chromatin restructuring, in both cases induction results in loss of nucleosome positioning. Glucose repression in the presence of inducer results in a specific pattern of partial positioning in the alcA and alcR promoters. Loss of nucleosome positioning depends absolutely on the AlcR protein and it is very unlikely to be a passive result of the induction of transcription. In an alcR loss-of-function background and in strains carrying mutations of the respective AlcR binding sites of the alcA and alcR promoters, nucleosomes are fully positioned under all growth conditions. Analysis of mutant AlcR proteins establishes that all domains needed for transcriptional activation and chromatin restructuring are included within the first 241 residues. The results suggest a two-step process, one step resulting in chromatin restructuring, a second one in transcriptional activation. Partial positioning upon glucose repression shows a specific pattern that depends on the CreA global repressor. An alcR loss-of-function mutation is epistatic to a creA loss-of function mutation, showing that AlcR does not act by negating a nucleosome positioning activity of CreA. PMID- 15813743 TI - The gene NCE103 (YNL036w) from Saccharomyces cerevisiae encodes a functional carbonic anhydrase and its transcription is regulated by the concentration of inorganic carbon in the medium. AB - Carbonic anhydrase (CA) catalyses the rapid interconversion between CO(2) and HCO(3) (-). Despite its wide distribution among living organisms, the presence of CA in fungi has been controversially discussed. Using mass spectrometric analysis of (18)O exchange from doubly labelled CO(2), we were able to measure CA activity in intact cells of Saccharomyces cerevisiae. Intracellular CA activity was lacking in the Deltance103 mutant, indicating that NCE103 encodes a functional CA. This was proven by overexpressing and purification of the NCE103 gene product showing a specific activity of around 6900 units per mg protein. Interestingly, the in vivo CA activity was 10-20 times higher in cells grown on low inorganic carbon (Ci; air containing 0.035% CO(2)) than in high-Ci cells (grown on 5% CO(2)). The enhanced CA activity of low-Ci cells was inducible after transferring high-Ci cells to air. Northern blot analysis revealed that that expression of NCE103 is transcriptionally regulated by low Ci which was also demonstrated by fusing the NCE103 promoter to beta-galactosidase as a reporter gene. Inactivation of NCE103 results in a high CO(2) requiring mutant indicating that a functional CA is an important prerequisite for S. cerevisiae to grow under low-Ci conditions. PMID- 15813744 TI - Regulation of azole drug susceptibility by Candida albicans protein kinase CK2. AB - Fluconazole resistance of the fungal pathogen Candida albicans can arise through several mechanisms, but the responsible genes and pathways are poorly understood. We report here that mutations in CKA2, identified through an insertional mutagenesis screen, confer fluconazole resistance. CKA2 and its homologue CKA1 specify catalytic subunits of protein kinase CK2. Although cka1 mutations have little effect on fluconazole resistance, CKA1 overexpression suppresses the fluconazole resistance of a cka2 mutant. This observation, along with synthetic cka1-cka2 interactions, argues that Cka1p and Cka2p carry out similar functions. cka2 mutants overexpress CDR1 and CDR2, two fluconazole efflux transporter genes, and a cdr1 mutation decreases resistance of a cka2 mutant, as expected if CDR1 and CDR2 overexpression is responsible for fluconazole resistance of the cka2 mutant. The protein phosphatase calcineurin is required for azole tolerance, and we find that the calcineurin inhibitor cyclosporin reverses fluconazole resistance of cka2 mutants. In addition, a mutation in CRZ1, which specifies a homologue of the Saccharomyces cerevisiae transcription factor that is a major target of calcineurin, suppresses fluconazole resistance of cka2 mutants. Expression analysis of Cka2p-responsive genes argues that Cka2p and Crz1p act through distinct mechanisms. Several clinical fluconazole-resistant isolates overexpress some Cka2p-responsive genes. We suggest that a Cka2p-dependent regulatory pathway is altered by clinically derived azole resistance mutations. PMID- 15813747 TI - The tetanus neurotoxin-sensitive and insensitive routes to and from the plasma membrane: fast and slow pathways? AB - Intracellular membrane trafficking in eukaryotes involves the budding of vesicles from a donor compartment, their translocation, and subsequent fusion with a target membrane. This last step has been shown to involve SNARE proteins, classified into two categories, vesicular (v)-SNAREs and target (t)-SNAREs. It is the pairing of v- and t-SNAREs that is responsible for bringing the lipid bilayers together for membrane fusion. Key to the discovery of SNAREs is the sensitivity of their neuronal synaptic prototypes, which mediate the release of neurotransmitters, to clostridial neurotoxins. In this review, we focus on tetanus neurotoxin-sensitive and tetanus neurotoxin-insensitive v-SNAREs, in particular synaptobrevin and cellubrevin, both tetanus neurotoxin-sensitive and Tetanus neurotoxin-Insensitive Vesicle-Associated Membrane Protein (TI-VAMP, also called VAMP7). The brevins are characterized by an RD sequence in the middle of their SNARE motif whereas TI-VAMP has an RG sequence. These two categories of exocytic v-SNAREs define two important routes to and from the plasma membrane: one sensitive, the other insensitive to tetanus neurotoxin. We also discuss the central role of the endosomal system that could be considered, as already suggested for Rab proteins, as a mosaic of v-SNAREs, thus raising the question of whether or not these two routes can merge, and if so, how and where. PMID- 15813746 TI - Intermediate filaments and vesicular membrane traffic: the odd couple's first dance? AB - During the last two decades, much attention has been focused on the regulation of membrane traffic by the actin and microtubule cytoskeletal networks. Their dynamic and polarized behavior and associated motors provide a logical framework from which architectural and movement cues can be communicated to organelles. The study of these cytoskeletal systems has been greatly aided by pharmacological agents. In contrast, intermediate filaments (IFs) have largely been neglected as a potential player in membrane traffic, both because a comprehensive pharmacology to perturb them does not exist and because they lack the intrinsic polarity and specific motors that make the other cytoskeletal systems attractive. In this review, we will discuss evidence suggesting that IFs may play roles in controlling organelle positioning and in membrane protein targeting. Furthermore, we will discuss potential mechanisms by which IFs may regulate the localization and function of organelles. PMID- 15813748 TI - Dissection of membrane dynamics of the ARF-guanine nucleotide exchange factor GBF1. AB - ADP-ribosylation factor (ARF)-facilitated recruitment of COP I to membranes is required for secretory traffic. The guanine nucleotide exchange factor GBF1 activates ARF and regulates ARF/COP I dynamics at the endoplasmic reticulum (ER) Golgi interface. Like ARF and coatomer, GBF1 peripherally associates with membranes. ADP-ribosylation factor and coatomer have been shown to rapidly cycle between membranes and cytosol, but the membrane dynamics of GBF1 are unknown. Here, we used fluorescence recovery after photobleaching to characterize the behavior of GFP-tagged GBF1. We report that GBF1 rapidly cycles between membranes and the cytosol (t1/2 is approximately 17 +/- 1 seconds). GBF1 cycles faster than GFP-tagged ARF, suggesting that in each round of association/dissociation, GBF1 catalyzes a single event of ARF activation, and that the activated ARF remains on membrane after GBF1 dissociation. Using three different approaches [expression of an inactive (E794K) GBF1 mutant, expression of the ARF1 (T31N) mutant with decreased affinity for GTP and Brefeldin A treatment], we show that GBF1 is stabilized on membranes when in a complex with ARF-GDP. GBF1 dissociation from ARF and membranes is triggered by its catalytic activity, i.e. the displacement of GDP and the subsequent binding of GTP to ARF. Our findings imply that continuous cycles of recruitment and dissociation of GBF1 to membranes are required for sustained ARF activation and COP I recruitment that underlies ER Golgi traffic. PMID- 15813749 TI - Requirement for microtubules and dynein motors in the earliest stages of peroxisome biogenesis. AB - Our aim was to determine the role of microtubules in the biogenesis of peroxisomes. Fusion experiments between human PEX16- and PEX1-mutant cells in the presence of nocodazol implied that microtubules were not required for import of proteins into the peroxisomal matrix after cell fusion complementation. We further studied the importance of microtubules in the early stages of peroxisome biogenesis following the microinjection complementation of PEX16-mutant cells. In the absence of nocodazol, nuclear microinjection of plasmids expressing EGFP-SKL and Pex16p in PEX16-mutant cells resulted in the accumulation of EGFP-SKL into newly formed peroxisomes. However, pretreatment of the cells with nocodazol, prior to microinjection, resulted in the inhibition of complementation of the PEX16 mutant and the cytosolic location of the EGFP-SKL. In addition, coexpression of a dominant-negative CC1 subunit of the dynein/dynactin motor complex resulted in the inability to complement PEX16-mutant cells. Both of these treatments resulted in the cytosolic localization of expressed Pex16p. Our results demonstrate that the formation of peroxisomes via the preperoxisomal compartment is dependent upon microtubules and minus-end-directed motor proteins and that the inhibition described above occurs at a step that precedes the association of Pex16p with the vesicles that would otherwise become the peroxisomes. PMID- 15813750 TI - Direct observation of rapid internalization and intracellular transport of sterol by macrophage foam cells. AB - Transport of the fluorescent cholesterol analog dehydroergosterol (DHE) from the plasma membrane was studied in J774 macrophages (Mphis) with normal and elevated cholesterol content. Cells were labeled with DHE bound to methyl-beta cyclodextrin. In J774, Mphis with normal cholesterol, intracellular DHE became enriched in recycling endosomes, but was not highly concentrated in the trans Golgi network or late endosomes and lysosomes. After raising cellular cholesterol by incubation with acetylated low-density lipoprotein (AcLDL), DHE was transported to lipid droplets, and less sterol was found in recycling endosomes. Transport of DHE to droplets was very rapid (t1/2 = 1.5 min after photobleaching) and did not require metabolic energy. In cholesterol-loaded J774 Mphis, the initial fraction of DHE in the plasma membrane was reduced, and rapid DHE efflux from the plasma membrane to intracellular organelles was observed. This rapid sterol transport was not related to plasma membrane vesiculation, as DHE did not become enriched in endocytic vesicles formed after sphingomyelinase C treatment of cells. When cells were incubated with DHE ester incorporated into AcLDL, fluorescence of the sterol was first found in punctate endosomes. After a chase, this DHE colocalized with transferrin in a distribution similar to cells labeled with DHE delivered by methyl-beta-cyclodextrin. Our results indicate that elevation of sterol levels in Mphis enhances transport of sterol from the plasma membrane by a non-vesicular pathway. PMID- 15813751 TI - The kinetics of phagosome maturation as a function of phagosome/lysosome fusion and acquisition of hydrolytic activity. AB - Professional phagocytes function at the hinge of innate and acquired immune responses by internalizing particulate material that is digested and sampled within the phagosome of the cell. Despite intense interest, assays to measure phagosome maturation remain insensitive and few in number. In this current study, we describe three novel assays that quantify important biological properties of the phagosome as it matures. One assay exploits fluorescence resonance energy transfer to quantify mixing of phagocytosed particles carrying a donor fluor with an acceptor fluor loaded previously into the lysosomes as a fluid phase marker. Two additional assays describe the functional maturation of the phagosome as a hydrolytic compartment following the degradation of specifically designed peptide and triglyceride fluorogenic substrates. The peptide substrate is preferentially cleaved by cysteine proteinases, and its degradation reflects proteinase delivery and activation within the acidifying phagosome. The fluorescence emission of the triglyceride analogue profiles the kinetics of triglyceride lipase activity within the phagosome. The fluorescence profiles of all three assays are modulated by known inhibitors of phagosome maturation, demonstrating the veracity, sensitivity and versatility of the assays. PMID- 15813752 TI - Translocation through the nuclear pore complex: selectivity and speed by reduction-of-dimensionality. AB - Translocation through the nuclear pore complex (NPC), a large transporter spanning the nuclear envelope, is a passive, diffusion-driven process, paradoxically enhanced by binding. To account for this mystery, several models have been suggested. However, recent experiments with modified NPCs make reconsideration necessary. Here, we suggest that nuclear transport receptors (NTRs) such as the karyopherins, in accordance with their peculiar boat-like structure, act as nanoscopic ferries transporting cargos through the NPC by sliding on a surface of phenylalanine glycine (FG) motifs. The dense array of FG motifs that covers the cytoplasmic filaments of the NPC is thought to continue on the wall of the large channel permeating the central framework of the NPC and on parts of the nuclear filaments to yield a coherent FG surface. Nuclear transport receptors are assumed to bind to the FG surface at filaments or at the channel entrance and then to rapidly search the FG surface by a two-dimensional random walk for the channel exit where they are released. The passage of neutral molecules is restricted to a narrow tube in the center of the central channel by a loose network of peptide chains. The model features virtual gating, is compatible with but not dependent on FG affinity gradients and tolerates deletions and transpositions of FG motifs. Implications of the model are discussed and tests are suggested. PMID- 15813753 TI - Self-directed learning--the importance of concepts and contexts. PMID- 15813754 TI - The importance of clinical outcomes in medical education research. PMID- 15813755 TI - Problem-based learning: let's not throw the baby out with the bathwater. PMID- 15813756 TI - Threats to the validity of clinical teaching assessments: what about rater error? PMID- 15813757 TI - How can medical students learn in a self-directed way in the clinical environment? Design-based research. AB - AIM: This study aimed to establish whether and under what conditions medical students can learn in a self-directed manner in the clinical environment. METHOD: A web-based learning management system brought 66 placement students, in a problem-based learning (PBL) medical curriculum, into closer touch with their clinical learning objectives and ways of achieving them. Free response comments from 16 of them during the 7 weeks they used it, transcripts of group discussions before and after the period of use, and responses from all 66 students to a questionnaire were analysed qualitatively. RESULTS: Students were rarely fully autonomous or subservient. They valued affective and pedagogic support, and relied on teachers to manage their learning environment. With support, they were motivated and able to choose how and when to meet their learning needs. The new system was a useful adjunct. CONCLUSIONS: Self-direction, interpreted literally, was a method of learning that students defaulted to when support and guidance were lacking. They found "supported participation" more valuable. Learning in the clinical environment was a social process with as many differences from, as similarities to, PBL. PMID- 15813758 TI - 'Sorry, it's my first time!' Will patients consent to medical students learning procedures? AB - CONTEXT: The clinical teaching of medical students is essential to medicine; however, medical students often may not inform patients of their inexperience. Hence patients do not have the opportunity to consent specifically to the procedures being performed by medical students. OBJECTIVES: The purpose of this study was to determine whether patients, when informed of the inexperience of a medical student, would still consent to the procedure. METHODS: Adult emergency department patients (114) were enrolled if they required one of the following procedures: sutures, intravenous (i.v.) access or splinting. Patients were first surveyed about his or her perceptions of medical student performing procedures. The first-year medical student then informed the patient of their inexperience (the number of procedures they had performed successfully). Finally, the patient was asked to consent to the student performing the procedure (i.e. "this is my first/third time suturing, may I suture you?") The main outcome measure was patient consent to the student performing the procedure. RESULTS: Only 48% of participants knew they could be the first patient on whom a medical student might perform a procedure. A total of 66% thought they should be told if a student was performing his or her first procedure on them. The majority of patients (90%) consented (69/80 i.v., 20/20 splints, 13/14 sutures). For 7 of the 12 refusals, it was the student's first time performing the procedure. CONCLUSION: Most patients will allow medical students to perform minor procedures, even when informed of the student's inexperience. PMID- 15813759 TI - The Self-Directed Learning Readiness Scale: a factor analysis study. AB - BACKGROUND: The practice of medicine demands that its physician practitioners are self-directed, life-long learners. The Self-Directed Learning Readiness Scale (SDLRS) intends to measure adults' readiness to engage in self-directed learning. PURPOSE: The present study assesses the underlying factor structure of the SDLRS for a sample of entering medical students. METHODS: Over a period of 6 years, 972 first year medical students at the Virginia Commonwealth University School of Medicine completed the SDLRS. To summarise the inter-relationships among variables, a principal axis factor analysis with oblique rotation was used on the 58 SDLRS items. A series of confirmatory factor analyses using LISREL 8.54 was performed to further examine the measurement model underlying the SDLRS. RESULTS: A 4-factor confirmatory model representing 4 correlated substantive factors and a reverse coding method factor fits these data well. CONCLUSIONS: Medical educators should hold limited expectations of the SDLRS to measure medical students' readiness to engage in self-directed learning. The definitions and theoretical assumptions that inform readiness for self-directed learning should be reconsidered. Alternative approaches to studying self-directed learning should be explored. PMID- 15813760 TI - Influence of new educational technology on problem-based learning at Harvard Medical School. AB - PURPOSE: Computers with 50-inch, wall-mounted plasma screens and broadband Internet access were installed in all small group tutorial rooms at Harvard Medical School. This study examines how the introduction of this educational technology impacted on the problem-based learning tutorials. METHOD: A total of 37 tutorial groups, stratified by year of student, were observed at separate timepoints (autumn 2002, spring 2003) to document the patterns of use of the technologies. Based on these observations, end-of-course surveys were developed and distributed to students and tutors. Observational field notes and open-ended survey responses were qualitatively analysed for themes. RESULTS: Using a 5-point rating scale, both students and tutors indicated that the technologies had a positive impact on their tutorials. In autumn 2002, plasma screens were used for an average of 17.8 and 22.1 minutes per 1-hour observation in Year 1 and 2 tutorials, respectively; in spring 2003, usage declined to 6.9 and 5.9 minutes, respectively. Resources utilised included Internet sites (54% total use time), PowerPoint presentations by students (22%), and course-specific postings (24%). Marked course-specific variation in usage was noted. Observational and survey data revealed that the technologies interrupted the flow of tutorial discussion. Students and tutors expressed concerns that the plasma screens might be altering the process of problem solving in the tutorials. CONCLUSIONS: Both students and tutors reported that the introduction of computers and wall-mounted plasma screens had impacted positively on tutorials. Questions were raised as to how this technology might alter tutorial dynamics. Further research will be needed to investigate these pedagogical concerns. PMID- 15813761 TI - Problem-based learning improves the academic performance of medical students in South Africa. AB - OBJECTIVES: To compare the academic performance of students on the previous, classical, discipline- and lecture-based, traditional curriculum with that of subsequent students who followed an innovative, problem- and community-based curriculum. METHODS: This was a retrospective study that analysed the records of students who enrolled on the doctor training programme between 1985 and 1995, and the records of students who graduated from the programme between 1989 and 2002. OUTCOMES: The educational outcomes assessed were the attrition and graduation rates on the traditional curriculum and those on the innovative curriculum. RESULTS: A total of 149 students on the traditional curriculum and 145 students on the innovative curriculum were studied. Overall, 23% of the traditional cohort as opposed to 10.3% of the innovative cohort dropped out of the course (P = 0.0041) and 55% of the traditional cohort as opposed to 67% of the innovative cohort graduated within the minimum period of 6 years (P < 0.001). The mean throughput period was 6.71 (0.09) years in the traditional cohort and 6.44 (0.07) years in the innovative cohort (P = 0.014) CONCLUSION: The introduction of the problem-based learning/community-based education (PBL/CBE) curriculum coincided with improved academic performance. The PBL/CBE approach to medical education may have contributed to this improvement. PMID- 15813762 TI - General competencies of problem-based learning (PBL) and non-PBL graduates. AB - INTRODUCTION: Junior doctors have reported shortcomings in their general competencies, such as organisational skills and teamwork. We explored graduates' perceptions of how well their training had prepared them for medical practice and in general competencies in particular. We compared the opinions of graduates from problem-based learning (PBL) and non-PBL schools, because PBL is supposed to enhance general competencies. METHOD: We analysed the responses of 1159 graduates from 1 PBL and 4 non-PBL schools to a questionnaire survey administered 18 months after graduation. RESULTS: Compared with their non-PBL colleagues, the PBL graduates gave higher ratings for the connection between school and work, their medical training and preparation for practice. According to the graduates, the most frequently used competencies with sufficient coverage during medical training were expert knowledge, profession-specific skills and communication skills. The majority of the PBL graduates, but less than half of the non-PBL graduates, indicated that communication skills had been covered sufficiently. All the graduates called for more curriculum attention on working with computers, planning and organisation, and leadership skills. More PBL graduates than non-PBL graduates indicated that they had learned profession-specific methods, communication skills and teamwork in medical school. DISCUSSION: Overall, the graduates appeared to be satisfied with their knowledge and skills. The results suggest that the PBL school provided better preparation with respect to several of the competencies. However, both PBL and non-PBL graduates identified deficits in their general competencies, such as working with computers and planning and organising work. These competencies should feature more prominently in undergraduate medical education. PMID- 15813763 TI - Electronic delivery of research summaries for academic generalist doctors: a randomised trial of an educational intervention. AB - OBJECTIVE: To determine whether weekly e-mails of structured, critically appraised summaries of new articles, within the format of a Weekly Browsing Journal Club (WBJC), would encourage use of evidence in practice. DESIGN: Randomised controlled trial. METHOD: A sample of 107 academic general internists were randomly assigned to receive either the WBJC (n = 54) or a health news website (n = 53) by e-mail on a weekly basis for 3 months. Participants completed a pre- and post-intervention questionnaire that measured their use of evidence in practice, attitudes about the role of evidence in practice, critical appraisal ability and reading habits. RESULTS: Of 107 participants, 90% completed the trial. At baseline, when faced with questions, they found answers on which to base their clinical decisions 67% of the time. Participants incorporated evidence into patient care for 60% of patients and read evidence to help make decisions for 25%. After 3 months, the groups did not differ in the use of evidence in practice. Changes in attitudes toward the role of evidence in practice and in self-perceived critical appraisal ability were also similar. The subjects spent 2.2 hours reading 4.3 journals per week. Weekly reading time decreased by 10 minutes in the WBJC group, but increased by 26 minutes in the control group (P = 0.02). At study completion, 82% of the WBJC group were satisfied with the intervention and 90% asked to continue. CONCLUSION: While doctors appreciated these summaries, which improved their reading efficiency, the intervention had little impact on their use of research evidence in practice. PMID- 15813764 TI - Do extended matching multiple-choice questions measure clinical reasoning? AB - PURPOSE: At the Faculty of Medicine at the Katholieke Universiteit Leuven, Belgium, we have developed a final examination that consists of extended matching multiple-choice questions. Extended matching questions (EMQs) originate from a case and have 1 correct answer within a list of at least 7 alternatives. If EMQs assess clinical reasoning, we can assume there will be a difference between the ways students and experienced doctors solve the problems within the questions. This study compared students' and residents' processes of solving EMQs. METHODS: Twenty final year students and 20 fourth or fifth year residents specialising in internal medicine solved 20 EMQs aloud. All questions concerned diagnosis or pathogenesis. Ten EMQs related to internal medicine and 10 questions to other medical disciplines. The session was audio-taped and transcribed. RESULTS: The residents correctly answered significantly more questions concerning internal medicine than did the students. Their reasoning was more "forward" and less "backward". No difference between residents and students was found for the other questions. The residents scored better on internal medicine than on the other questions. They used more backward and less forward reasoning when solving the other questions than they did with the internal medicine questions. The better half of the respondents used significantly more forward and less backward reasoning than did the poorer half. CONCLUSION: In accordance with the literature, medical expertise was characterised by forward reasoning, whereas outside their area of expertise, the subjects switched over to backward reasoning. It is possible to assess processes of clinical reasoning using EMQs. PMID- 15813765 TI - Research in clinical reasoning: past history and current trends. AB - BACKGROUND: Research in clinical reasoning has been conducted for over 30 years. Throughout this time there have been a number of identifiable trends in methodology and theory. PURPOSE: This paper identifies three broad research traditions, ordered chronologically, are: (a) attempts to understand reasoning as a general skill--the "clinical reasoning" process; (b) research based on probes of memory--reasoning related to the amount of knowledge and memory; and (c) research related to different kinds of mental representations--semantic qualifiers, scripts, schemas and exemplars. RESULTS AND CONCLUSIONS: Several broad themes emerge from this review. First, there is little evidence that reasoning can be characterised in terms of general process variables. Secondly, it is evident that expertise is associated, not with a single basic representation but with multiple coordinated representations in memory, from causal mechanisms to prior examples. Different representations may be utilised in different circumstances, but little is known about the characteristics of a particular situation that led to a change in strategy. IMPLICATIONS: It becomes evident that expertise lies in the availability of multiple representations of knowledge. Perhaps the most critical aspect of learning is not the acquisition of a particular strategy or skill, nor is it the availability of a particular kind of knowledge. Rather, the critical element may be deliberate practice with multiple examples which, on the hand, facilitates the availability of concepts and conceptual knowledge (i.e. transfer) and, on the other hand, adds to a storehouse of already solved problems. PMID- 15813766 TI - Conversation analysis, doctor-patient interaction and medical communication. AB - INTRODUCTION: This paper introduces medical educators to the field of conversation analysis (CA) and its contributions to the understanding of the doctor-patient relationship. THE CONVERSATION ANALYSIS APPROACH: Conversation analysis attempts to build bridges both to the ethnographic and the coding and quantitative studies of medical interviews, but examines the medical interview as an arena of naturally occurring interaction. This implies distinctive orientations and issues regarding the analysis of doctor-patient interaction. We discuss the CA approach by highlighting 5 basic features that are important to the enterprise, briefly illustrating each issue with a point from research on the medical interview. These features of conversation analytic theory and method imply a systematic approach to the organisation in interaction that distinguishes it from studies that rely on anecdote, ethnographic inquiry or the systematic coding of utterances. CONVERSATION ANALYSIS AND THE MEDICAL INTERVIEW: We then highlight recent CA studies of the "phases" of the internal medicine clinic and the implications of these studies for medical education. We conclude with suggestions for how to incorporate CA into the medical curriculum. It fits with biopsychosocial, patient-centred and relationship-centred approaches to teaching about medical communication. PMID- 15813767 TI - The use (and abuse) of the term "portfolio". PMID- 15813768 TI - Lumbar puncture experience among medical school graduates: the need for formal procedural skills training. PMID- 15813769 TI - Hakan Tegelstrom ('HaTe'): naturalist and geneticist 2 August 1947-20 March 2004. PMID- 15813770 TI - Substantial genetic substructuring in southeastern and alpine Australia revealed by molecular phylogeography of the Egernia whitii (Lacertilia: Scincidae) species group. AB - Palaeoclimatic events and biogeographical processes since the mid-Tertiary are believed to have strongly influenced the evolution and distribution of the terrestrial vertebrate fauna of southeastern Australia. We examined the phylogeography of the temperate-adapted members of the Egernia whitii species group, a group of skinks that comprise both widespread low- to mid-elevation (E. whitii) and montane-restricted species (Egernia guthega, Egernia montana), in order to obtain important insights into the influence of past biogeographical processes on the herpetofauna of southeastern Australia. Sequence data were obtained from all six temperate-adapted species within the E. whitii species group, and specifically from across the distributional ranges of E. whitii, E. guthega and E. montana. We targeted a fragment of the ND4 mitochondrial gene (696 bp) and analysed the data using maximum likelihood and Bayesian methods. Our data reveal a deep phylogeographical break in the east Gippsland region of Victoria between 'northern' (Queensland, New South Wales, Australian Capital Territory) and 'southern' (Victoria, Tasmania, South Australia) populations of E. whitii. This divergence appears to have occurred during the late Miocene-Pliocene, with the Gippsland basin possibly forming a geographical barrier to dispersal. Substantial structuring within both the 'northern' and the 'southern' clades is consistent with the effects of Plio-Pleistocene glacial-interglacial cycles. Pleistocene glacial cycles also appear to have shaped the phylogeographical patterns observed in the alpine species, E. guthega and E. montana. We used our results to examine the biogeographical process that led to the origin and subsequent diversification of the lowland and alpine herpetofauna of southeastern Australia. PMID- 15813771 TI - A multilocus population genetic survey of the greater sage-grouse across their range. AB - The distribution and abundance of the greater sage-grouse (Centrocercus urophasianus) have declined dramatically, and as a result the species has become the focus of conservation efforts. We conducted a range-wide genetic survey of the species which included 46 populations and over 1000 individuals using both mitochondrial sequence data and data from seven nuclear microsatellites. Nested clade and structure analyses revealed that, in general, the greater sage-grouse populations follow an isolation-by-distance model of restricted gene flow. This suggests that movements of the greater sage-grouse are typically among neighbouring populations and not across the species, range. This may have important implications if management is considering translocations as they should involve neighbouring rather than distant populations to preserve any effects of local adaptation. We identified two populations in Washington with low levels of genetic variation that reflect severe habitat loss and dramatic population decline. Managers of these populations may consider augmentation from geographically close populations. One population (Lyon/Mono) on the southwestern edge of the species' range appears to have been isolated from all other greater sage-grouse populations. This population is sufficiently genetically distinct that it warrants protection and management as a separate unit. The genetic data presented here, in conjunction with large-scale demographic and habitat data, will provide an integrated approach to conservation efforts for the greater sage grouse. PMID- 15813772 TI - Genetic analysis of the breeding system of an invasive subterranean termite, Reticulitermes santonensis, in urban and natural habitats. AB - Reticulitermes santonensis is a subterranean termite that invades urban areas in France and elsewhere where it causes damage to human-built structures. We investigated the breeding system, colony and population genetic structure, and mode of dispersal of two French populations of R. santonensis. Termite workers were sampled from 43 and 31 collection points, respectively, from a natural population in west-central France (in and around the island of Oleron) and an urban population (Paris). Ten to 20 workers per collection point were genotyped at nine variable microsatellite loci to determine colony identity and to infer colony breeding structure. There was a total of 26 colonies, some of which were spatially expansive, extending up to 320 linear metres. Altogether, the analysis of genotype distribution, F-statistics and relatedness coefficients suggested that all colonies were extended families headed by numerous neotenics (nonwinged precocious reproductives) probably descended from pairs of primary (winged) reproductives. Isolation by distance among collection points within two large colonies from both populations suggested spatially separated reproductive centres with restricted movement of workers and neotenics. There was a moderate level of genetic differentiation (F(ST) = 0.10) between the Oleron and Paris populations, and the number of alleles was significantly higher in Oleron than in Paris, as expected if the Paris population went through bottlenecks when it was introduced from western France. We hypothesize that the diverse and flexible breeding systems found in subterranean termites pre-adapt them to invade new or marginal habitats. Considering that R. santonensis may be an introduced population of the North American species R. flavipes, a breeding system consisting primarily of extended family colonies containing many neotenic reproductives may facilitate human-mediated spread and establishment of R. santonensis in urban areas with harsh climates. PMID- 15813773 TI - Nonequilibrium conditions following landscape rearrangement: the relative contribution of past and current hydrological landscapes on the genetic structure of a stream-dwelling fish. AB - Interpreting patterns of population structure in nature is often challenging, especially in dynamic landscapes where population genetic connectivity evolves over time. In this study, we document the absence of migration-drift equilibrium in a stream-dwelling euryhaline fish resulting from past fine-scale drainage rearrangements and evaluate the relative contribution of past and current hydrological landscapes on observed population structure. Based on allelic variation at nine microsatellite loci, genetic relationships among 12 populations of brook charr, Salvelinus fontinalis, from Gros Morne National Park of Canada (GMNP, Newfoundland, Canada) did not reflect current stream hierarchical structure. In addition, we observed no correlation between population differentiation and contemporary landscape features (waterway distance and sums of altitudinal differences). Instead, population relationships were consistent with historical hydrological structure predicted a priori based on geomorphological and biogeographical evidences. Also, population differentiation was strongly correlated with inferred historical landscape features. Contemporary barriers have apparently preserved the signature of past genetic connectivity by constraining gene flow. Based on the relationships between population differentiation and current and past landscape features at various spatial scales, we suggest that brook charr genetic diversity in GMNP is mostly the result of small distance migrations at the time of colonization and subsequent differentiation through drift. This study highlights the potential of approaching landscapes from a combination of contemporary and historical perspectives when interpreting nonequilibrium population structures resulting from landscape rearrangement. PMID- 15813774 TI - Chloroplast microsatellites: measures of genetic diversity and the effect of homoplasy. AB - Chloroplast microsatellites have been widely used in population genetic studies of conifers in recent years. However, their haplotype configurations suggest that they could have high levels of homoplasy, thus limiting the power of these molecular markers. A coalescent-based computer simulation was used to explore the influence of homoplasy on measures of genetic diversity based on chloroplast microsatellites. The conditions of the simulation were defined to fit isolated populations originating from the colonization of one single haplotype into an area left available after a glacial retreat. Simulated data were compared with empirical data available from the literature for a species of Pinus that has expanded north after the Last Glacial Maximum. In the evaluation of genetic diversity, homoplasy was found to have little influence on Nei's unbiased haplotype diversity (H(E)) while Goldstein's genetic distance estimates (D2sh) were much more affected. The effect of the number of chloroplast microsatellite loci for evaluation of genetic diversity is also discussed. PMID- 15813775 TI - Genetic structure and history of populations of the deep-sea fish Helicolenus dactylopterus (Delaroche, 1809) inferred from mtDNA sequence analysis. AB - Helicolenus dactylopterus is an Atlantic benthopelagic fish species inhabiting high-energy habitats on continental slopes, seamounts and islands. Partial sequences of the mitochondrial control region (D-loop) and cytochrome b (cyt b) were used to test the hypothesis that H. dactylopterus disperses between continental margin, island and seamount habitats on intraregional, regional and oceanic scales in the North Atlantic. Individuals were collected from five different geographical areas: Azores, Madeira, Portugal (Peniche), Cape Verde and the northwest Atlantic. D-loop (415 bp) and cyt b (423 bp) regions were partially sequenced for 208 and 212 individuals, respectively. Analysis of variation among mitochondrial DNA sequences based on pairwise F-statistics and AMOVA demonstrated marked genetic differentiation between populations in different geographical regions specifically the Mid-Atlantic Ridge (Azores)/northeast Atlantic (Portugal, Madeira) compared to populations around the Cape Verde Islands and in the northwest Atlantic. Some evidence of intraregional genetic differentiation between populations was found. Minimum-spanning network analysis revealed star shaped patterns suggesting that populations had undergone expansion following bottlenecks and/or they have been colonized by jump dispersal events across large geographical distances along pathways of major ocean currents. Mismatch distribution analysis indicated that Azores and northwest Atlantic populations fitted a model of historical population expansion following a bottleneck/founder event estimated to be between 0.64 and 1.2 million years ago (Ma). PMID- 15813776 TI - F-statistics under alternation of sexual and asexual reproduction: a model and data from schistosomes (platyhelminth parasites). AB - Accurate inferences on population genetics data require a sound underlying theoretical null model. Nearly nothing is known about the gene dynamics of organisms with complex life cycles precluding any biological interpretation of population genetics parameters. In this article, we used an infinite island model to derive the expectations of those parameters for the life cycle of a dioecious organism obligatorily alternating sexual and asexual reproductions as it is the case for schistosomes (plathyhelminth parasites). This model allowed us to investigate the effects of the degree of mixing among individuals coming from different subpopulations at each new generation (represented in the model by the migration rates before and after clonal reproductions) and the variance in the reproductive success of individuals during the clonal phase. We also consider the effects of different migration rates and degrees of clonal reproductive skew between male and female individuals. Results show that the variance in the reproductive success of clones is very important in shaping the distribution of the genetic variability both within and among subpopulations. Thus, higher variance in the reproductive success of clones generates heterozygous excesses within subpopulations and also increases genetic differentiation between them. Migration occurring before and after asexual reproduction has different effects on the patterns of F(IS) and F(ST). When males and females display different degrees of reproductive skew or migration rates, we observe differences in their respective population genetic structure. While results of the model apply to any organism alternating sexual and clonal reproductions (e.g. all parasitic trematodes, many plants, and all aphididae), we finally confront some of these theoretical expectations to empirical data from Schistosoma mansoni infecting Rattus rattus in Guadeloupe. PMID- 15813777 TI - Genetic structure in the coral-reef-associated Banggai cardinalfish, Pterapogon kauderni. AB - In this study, we used 11 polymorphic microsatellite loci to show that oceanic distances as small as 2-5 km are sufficient to produce high levels of population genetic structure (multilocus F(ST) as high as 0.22) in the Banggai cardinalfish (Pterapogon kauderni), a heavily exploited reef fish lacking a pelagic larval dispersal phase. Global F(ST) among all populations, separated by a maximum distance of 203 km, was 0.18 (R(ST) = 0.35). Moreover, two lines of evidence suggest that estimates of F(ST) may actually underestimate the true level of genetic structure. First, within-locus F(ST) values were consistently close to the theoretical maximum set by the average within-population heterozygosity. Second, the allele size permutation test showed that R(ST) values were significantly larger than F(ST) values, indicating that populations have been isolated long enough for mutation to have played a role in generating allelic variation among populations. The high level of microspatial structure observed in this marine fish indicates that life history traits such as lack of pelagic larval phase and a good homing ability do indeed play a role in shaping population genetic structure in the marine realm. PMID- 15813778 TI - Regionally isolated populations of an imperiled Caribbean coral, Acropora palmata. AB - The movements of larvae between marine populations are difficult to follow directly and have been the subject of much controversy, especially in the Caribbean. The debate centres on the degree to which populations are demographically open, such that depleted populations can be replenished by recruitment from distant healthy populations, or demographically closed and thus in need of local management. Given the depressed state of many tropical reef populations, the understanding of these movements now bears critically on the number, placement, and size of marine reserves. Most genetic analyses assume that dispersal patterns have been stable for thousands of generations, thus they commonly reflect past colonization histories more than ongoing dispersal. Recently developed multilocus genotyping approaches, however, have the demonstrated ability to detect both migration and population isolation over far shorter timescales. Previously, we developed five microsatellite markers and demonstrated them to be both Mendelian and coral-specific. Using these markers and Bayesian analyses, we show here that populations of the imperiled reef building coral, Acropora palmata, have experienced little or no recent genetic exchange between the western and the eastern Caribbean. Puerto Rico is identified as an area of mixing between the two subregions. As a consequence of this regional isolation, populations in the western and eastern Caribbean should have the potential to adapt to local conditions and will require population-specific management strategies. PMID- 15813779 TI - Low level of gene flow from cultivated beets (Beta vulgaris L. ssp. vulgaris) into Danish populations of sea beet (Beta vulgaris L. ssp. maritima (L.) Arcangeli). AB - Gene flow from sugar beets to sea beets occurs in the seed propagation areas in southern Europe. Some seed propagation also takes place in Denmark, but here the crop-wild gene flow has not been investigated. Hence, we studied gene flow to sea beet populations from sugar beet lines used in Danish seed propagation areas. A set of 12 Danish, two Swedish, one French, one Italian, one Dutch, and one Irish populations of sea beets, and four lines of sugar beet were analysed. To evaluate the genetic variation and gene flow, eight microsatellite loci were screened. This analysis revealed hybridization with cultivated beet in one of the sea beet populations from the centre of the Danish seed propagation area. Triploid hybrids found in this population were verified with flow cytometry. Possible hybrids or introgressed plants were also found in the French and Italian populations. However, individual assignment test using a Bayesian method provided 100% assignment success of diploid individuals into their correct subspecies of origin, and a Bayesian Markov chain Monte Carlo (MC MC) approach revealed clear distinction of individuals into groups according to their subspecies of origin, with a zero level of genetic admixture among subspecies. This underlines that introgression beyond the first hybridization is not extensive. The overall pattern of genetic distance and structure showed that Danish and Swedish sea beet populations were closely related to each other, and they are both more closely related to the population from Ireland than to the populations from France, the Netherlands, and Italy. PMID- 15813780 TI - Genetic continuity of brood-parasitic indigobird species. AB - Speciation in brood-parasitic indigobirds (genus Vidua) is a consequence of behavioural imprinting in both males and females. Mimicry of host song by males and host fidelity in female egg laying result in reproductive isolation of indigobirds associated with a given host species. Colonization of new hosts and subsequent speciation require that females occasionally lay eggs in the nests of novel hosts but the same behaviour may lead to hybridization when females parasitize hosts already associated with other indigobird species. Thus, retained ancestral polymorphism and ongoing hybridization are two alternative explanations for the limited genetic differentiation among indigobird species. We tested for genetic continuity of indigobird species using mitochondrial sequences and nuclear microsatellite data. Within West Africa and southern Africa, allopatric populations of the same species are generally more similar to each other than to sympatric populations of different species. Likewise, a larger proportion of genetic variation is explained by differences between species than by differences between locations in alternative hierarchical AMOVAS, suggesting that the rate of hybridization is not high enough to homogenize sympatric populations of different species or prevent genetic differentiation between species. Broad sharing of genetic polymorphisms among species, however, suggests that some indigobird species trace to multiple host colonization events in space and time, each contributing to the formation of a single interbreeding population bound together by songs acquired from the host species. PMID- 15813781 TI - Northwestern song sparrow populations show genetic effects of sequential colonization. AB - Two genetic consequences are often considered evidence of a founder effect: substantial loss in genetic diversity and rapid divergence between source and founder populations. Single-step founder events have been studied for these effects, but with mixed results, causing continued controversy over the role of founder events in divergence. Experiments of serial bottlenecks have shown losses of diversity, increased divergence, and rapid behavioural changes possibly leading to reproductive isolation between source and final populations. The few studies conducted on natural, sequentially founded systems show some evidence of these effects. We examined a natural vertebrate system of sequential colonization among northwestern song sparrows (Melospiza melodia). This system has an effectively linear distribution, it was probably colonized within the last 10,000 years, there are morphological and behavioural differences among populations, and the westernmost populations occur in atypical habitats for the species. Eight microsatellite loci from eight populations in Alaska and British Columbia (n = 205) showed stepwise loss of genetic diversity, genetic evidence for strong population bottlenecks, and increased population divergence. The endpoint population on Attu Island has extremely low diversity (H(E) = 0.18). Our study shows that sequential bottlenecks or founder events can have powerful genetic effects in reducing diversity, possibly leading to rapid evolutionary divergence. PMID- 15813782 TI - Phylogeographic structure and postglacial evolutionary history of water voles (Arvicola terrestris) in the United Kingdom. AB - The phylogeographic pattern of mitochondrial DNA variation in water voles (Arvicola terrestris) from 57 localities across the United Kingdom and representative samples from Spain, France, Switzerland and Finland was determined from sequence variation in the central portion of the control region. Twenty seven different haplotypes were resolved which formed two distinct phylogenetic clades. This major division separated haplotypes found in Scotland from those found in England and Wales. Nested clade analysis of haplotypes indicated that such a division was a consequence of allopatric fragmentation. The haplotypes found in Switzerland, France and Spain clustered with Scottish haplotypes, whereas the haplotype from Finland clustered with the English/Welsh haplotypes. These patterns indicate that contemporary Scottish populations are derived from an Iberian glacial refugium, whereas English and Welsh populations are derived from an eastern European refugium. As such, the postglacial recolonization of the United Kingdom must have involved two colonization events, either in different localities with no subsequent contact, or as two waves separated over time, with the second wave of colonizers displacing the first. An analysis of molecular variance (AMOVA) identified significant population genetic divergence within both the major clades, indicative of restricted gene flow and regional population isolation. The implications of both phylogeographical and population genetic structure are discussed in context with the conservation of water voles in Britain. PMID- 15813783 TI - Postglacial expansion of the southern red-backed vole (Clethrionomys gapperi) in North America. AB - Dynamic climatic oscillations of the Pleistocene dramatically changed the distributions of high latitude species. Molecular investigations of a variety of organisms show that processes of postglacial colonization of boreal regions were more complex than initially thought. Phylogeographical and coalescent analyses were conducted on partial sequences of the cytochrome b gene (600 bp) from 64 individuals of Clethrionomys gapperi from North Carolina, Pennsylvania, Minnesota, Idaho, Washington, British Columbia, Northwest Territories, and Alaska to test hypotheses relating to Pleistocene refugia and postglacial colonization routes. Three divergent clades (east, west, central) were identified with highest net divergence (dA = 5.2%) between the eastern and western clades. Populations from the recently deglaciated higher latitudes of Canada and Alaska are closely related to lower latitude populations of the central clade (dA = 1.2%) suggesting recent expansion from this midwestern region. No representatives from the east or west clade were found at latitudes higher than 50 degrees N, indicating that postglacial colonization occurred through a midcontinental route. The high latitude population from the Northwest Territories exhibited demographic patterns and genetic diversity consistent with a stable noncolonizing population. This population is found near the Mackenzie range, where the two continental ice sheets were believed to have coalesced. Molecular variation observed in this population may be the result of leading edge population diversifying in the continental corridor or may reflect the signal of a high latitude refugial population. PMID- 15813784 TI - Phylogeographic analysis of mitochondrial gene flow and introgression in the salamander, Plethodon shermani. AB - Plethodon shermani comprises a series of geographically disjunct populations occupying high-elevation mountain isolates. These populations hybridize at their borders with salamanders of the Plethodon glutinosus species complex, and past range expansions inferred from Pleistocene climatic cycles may have increased the possible genetic interactions between P. shermani and species of the P. glutinosus complex. Because mitochondrial DNA haplotypes often show introgression across species borders, we survey mtDNA variation for evidence of past and ongoing genetic interactions between P. shermani, its close relative Plethodon cheoah, and species of the P. glutinosus complex. Ongoing hybridization with the P. glutinosus-complex species Plethodon teyahalee is accompanied by extensive mitochondrial introgression in some Unicoi populations of P. shermani, but it has little genetic impact on P. shermani populations outside hybrid zones at three other isolates (Tusquitee, Wayah Bald, Standing Indian). Some Unicoi populations of P. shermani exhibit mtDNA evidence of past hybridization with diverse lineages from P. aureolus and P. glutinosus. The Tusquitee isolate of P. shermani is also characterized by mtDNA haplotypes most closely related to Plethodon aureolus and P. glutinosus, presumably introduced by past genetic contact with these species or with introgressed populations of Unicoi P. shermani. The mtDNA variation in sampled populations of the Wayah Bald and Standing Indian isolates of P. shermani appears largely unaffected by ongoing hybridization. Principal components analyses of allozymic data indicate that P. shermani isolates collectively form a genetically homogeneous unit clearly demarcated from species with which they have had current or past genetic interactions. Rapid mtDNA introgression associated with transient contacts between P. shermani and other species permits a fine level resolution of evolutionary lineages not evident from allozymic data. PMID- 15813785 TI - Genetic spatial structure of European common hamsters (Cricetus cricetus)--a result of repeated range expansion and demographic bottlenecks. AB - The spatial genetic structure of common hamsters (Cricetus cricetus) was investigated using three partial mitochondrial (mt) genes and 11 nuclear microsatellite loci. All marker systems revealed significant population differentiation across Europe. Hamsters in central and western Europe belong largely to two allopatric mitochondrial lineages south and northwest of the Carpathian and Sudetes. The southern group, 'Pannonia', comprises populations inside the Carpathian basin (Czech Republic, Hungary) while the second group, 'North', includes hamsters from Belgium, the Netherlands, France, and Germany. Isolation of the lineages is maintained by a combination of geographical and ecological barriers. Both main phylogeographical groups show signs of further subdivision. North is separated into highly polymorphic central German and less polymorphic western populations, which most likely split during late glacial expansion (15,000-10,000 bp). Clock estimates based on haplotype distributions predict a divergence of the two major lineages 85,000-147,000 bp. Expansion times fall during the last glaciation (115,000-10,000 bp) corroborating fossil data, which identify Cricetus cricetus as characteristic of colder climatic phases. Despite the allopatry of mt haplotypes, there is an overlap of nuclear microsatellite alleles between phylogeographical units. Although there are strong evidence that Pannonian hamsters have persisted inside the Carpathian basin over the last 50,000 years, genetic differentiation among European hamsters has mainly been caused by immigration from different eastern refugia. Possible source populations are likely to be found in the Ukrainian and the southern Russian plains--core areas of hamster distribution. From there, hamsters have repeatedly expanded during the Quaternary. PMID- 15813786 TI - Phylogeography of an east Australian wet-forest bird, the satin bowerbird (Ptilonorhynchus violaceus), derived from mtDNA, and its relationship to morphology. AB - Australian wet forests have undergone a contraction in range since the mid Tertiary, resulting in a fragmented distribution along the east Australian coast incorporating several biogeographical barriers. Variation in mitochondrial DNA and morphology within the satin bowerbird was used to examine biogeographical structure throughout almost the entire geographical extent of these wet forest fragments. We used several genetic analysis techniques, nested clade and barrier analyses, that use patterns inherent in the data to describe the spatial structuring. We also examined the validity of the two previously described satin bowerbird subspecies that are separated by well-defined biogeographical barriers and tested existing hypotheses that propose divergence occurs within each subspecies across two other barriers, the Black Mountain corridor and the Hunter Valley. Our data showed that the two subspecies were genetically and morphologically divergent. The northern subspecies, found in the Wet Tropics region of Queensland, showed little divergence across the Black Mountain corridor, a barrier found to be significant in other Wet Tropics species. Biogeographical structure was found through southeastern Australia; three geographically isolated populations showed genetic differentiation, although minimal divergence was found across the proposed Hunter Valley barrier. A novel barrier was found separating inland and coastal populations in southern New South Wales. Little morphological divergence was observed within subspecies, bar a trend for birds to be larger in the more southerly parts of the species' range. The results from both novel and well-established genetic analyses were similar, providing greater confidence in the conclusions about spatial divergence and supporting the validity of these new techniques. PMID- 15813787 TI - Allopatric origins of sympatric brook charr populations: colonization history and admixture. AB - Natural selection is presumed to be the driving force behind the occurrence of phenotypically and genetically divergent populations in sympatry within many north temperate freshwater fishes. If, however, these populations have different ancestral origins, history could also contribute to their divergence. We previously found evidence for the role of selection in the evolution of divergent outflow and inflow breeding populations of migratory brook charr (Salvelinus fontinalis) inhabiting postglacial Mistassini Lake (Quebec, Canada). Here, we show that these populations do not have a common origin, through the use of admixture and spatial analyses with seven microsatellite loci. Divergent populations clustered into two different population groups when compared to samples from surrounding drainages, although inflow populations appeared to be more admixed between the two population groups than the outflow population. These results are noteworthy since outflow and inflow populations were monomorphic at mitochondrial DNA (338-bp sequence of the control region) and are only moderately differentiated (mean F(ST) = 0.10). Colonization by two ancestral populations was also consistent with known outflow direction changes throughout lake formation. In addition to providing insight into how phenotypic divergence in sympatry may have been affected by the nature (i.e. timing and direction) of colonization of ancestral populations, our results also suggest that ancestral populations may have differed in their ability to colonize certain lake habitats. PMID- 15813788 TI - Similar mating and sperm displacement patterns in two highly divergent D. simulans populations from Africa and Europe. AB - The frequency of remating in Drosophila melanogaster is affected by both genetic and ecological factors. We studied the remating behaviour in one European (Italy) and one African (Uganda) Drosophila simulans population using six highly polymorphic microsatellite markers. Despite that the populations were genetically distinct (F(ST) = 0.18) and originated from very dissimilar ecological settings with different population densities, we inferred a very similar mating pattern. The remating parameter alpha was similar in both populations (a = 1.3-1.4). No more than two distinct paternal genotypes per family were detected in each population. PMID- 15813789 TI - Geographic variation of multiple paternity in the American lobster, Homarus americanus. AB - We studied the frequency of multiple paternity for American lobster (Homarus americanus) at three Canadian sites differing in exploitation rate and mean adult size. The probability of detecting multiple paternity using four microsatellite loci and 100 eggs per female was in excess of 99% under various scenarios of paternal contribution. Overall, 13% of the 108 examined females carried a clutch sired by two or three males. Multiple paternity was observed at the two most exploited sites (11% at Magdalen Islands and 28% at Grand Manan Island), whereas single paternity only was observed at the least exploited site (Anticosti Island). Within populations females with a clutch sired by more than one male tended to be smaller than females with a clutch sired by a single male. Based on these and other findings, we postulate a link between female promiscuity and sperm limitation in the American lobster. PMID- 15813790 TI - Unrelated queens coexist in colonies of the termite Macrotermes michaelseni. AB - Relatedness increases the likelihood of cooperation within colonies of social insects. Polygyny, the coexistence of numerous reproductive females (queens) in a colony, is common in mature colonies of the termite Macrotermes michaelseni. In this species, polygyny results from pleometrosis and from several female alates that jointly found a new colony. To explain this phenomenon, it was suggested that only related females cooperate and survive during maturation of colonies. Using multilocus fingerprints as well as microsatellites, we showed that nestmate queens in mature colonies are unrelated. Furthermore, we found that all nestmate queens contributed to the production of steriles. Even in mature colonies, several matrilines of steriles coexist within a colony. Although genetic diversity within colonies may increase the likelihood of conflicts, high genetic diversity may be important for foraging, colony growth, and resistance to disease and parasites. PMID- 15813791 TI - Sexual selection for male dominance reduces opportunities for female mate choice in the European bitterling (Rhodeus sericeus). AB - Sexual selection involves two main mechanisms: intrasexual competition for mates and intersexual mate choice. We experimentally separated intrasexual (male-male interference competition) and intersexual (female choice) components of sexual selection in a freshwater fish, the European bitterling (Rhodeus sericeus). We compared the roles of multiple morphological and behavioural traits in male success in both components of sexual competition, and their relation to male reproductive success, measured as paternity of offspring. Body size was important for both female choice and male-male competition, though females also preferred males that courted more vigorously. However, dominant males often monopolized females regardless of female preference. Subordinate males were not excluded from reproduction and sired some offspring, possibly through sneaked ejaculations. Male dominance and a greater intensity of carotenoid-based red colouration in their iris were the best predictors of male reproductive success. The extent of red iris colouration and parasite load did not have significant effects on female choice, male dominance or male reproductive success. No effect of parasite load on the expression of red eye colouration was detected, though this may have been due to low parasite prevalence in males overall. In conclusion, we showed that even though larger body size was favoured in both intersexual and intrasexual selection, male-male interference competition reduced opportunities for female choice. Females, despite being choosy, had limited control over the paternity of their offspring. Our study highlights the need for reliable measures of male reproductive success in studies of sexual selection. PMID- 15813792 TI - Social parasitism in fire ants (Solenopsis spp.): a potential mechanism for interspecies transfer of Wolbachia. AB - One possible mechanism for interspecific transfer of Wolbachia is through the intimate contact between parasites and their hosts. We surveyed 10 species of fly parasitoids (Pseudacteon spp.) and one inquiline social parasite, Solenopsis daguerrei, for the presence and sequence identity (wsp gene) of Wolbachia. Two Wolbachia variants infecting S. daguerrei were identical to known variants infecting the two common ant host species, Solenopsis invicta and Solenopsis richteri, suggesting possible transfers of Wolbachia between this parasite and their hosts have occurred. Our data also revealed an unexpectedly high diversity of Wolbachia variants within S. daguerrei: up to eight variants were found within each individual, which, to our knowledge, is the highest reported number of Wolbachia variants infecting a single individual of any host species. PMID- 15813793 TI - On the origins of extreme mycorrhizal specificity in the Monotropoideae (Ericaceae): performance trade-offs during seed germination and seedling development. AB - Fungal-induced seed germination is a phenomenon characteristic of mycorrhizal plants that produce dust-like seeds with only minimal nutritional reserves. In such systems, fungi trigger germination and/or subsidize development. We studied mycorrhizal germination in relation to mycorrhizal specificity in the Monotropoideae, a lineage of dust-seeded non-photosynthetic plants that are dependent upon ectomycorrhizal fungi of forest trees. A total of 1695 seed packets, each containing two to five compartments with seeds from different sources, were buried for up to 2 years near known ectomycorrhizal fungi in six different native forest locations. Upon harvest, seedlings were analysed by cultivation-independent molecular methods to identify their mycorrhizal fungi. We report that (i) germination is only induced by the same fungus that associates with mature plants or by closely related congeners; (ii) seedlings associated with the latter fungi develop less than those associated with maternal fungal species in most settings; and (iii) exceptions to this pattern occur in allopatric settings, where novel plant-fungal associations can result in the greatest seedling development. We interpret these results as evidence of performance trade-offs between breadth of host range and rate of development. We propose that in conjunction with host-derived germination cues, performance trade offs can explain the extreme mycorrhizal specificity observed at maturity. The allopatric exceptions support the idea that performance trade-offs may be based on a coevolutionary arms race and that host range can be broadened most readily when naive fungal hosts are encountered in novel settings. PMID- 15813794 TI - Transposable element polymorphism of Wolbachia in the mosquito Culex pipiens: evidence of genetic diversity, superinfection and recombination. AB - Wolbachia is a group of maternally inherited endosymbiotic bacteria that infect and induce cytoplasmic incompatibility (CI) in a wide range of arthropods. In contrast to other species, the mosquito Culex pipiens displays an extremely high number of CI types suggesting differential infection by multiple Wolbachia strains. Attempts so far failed to detect Wolbachia polymorphism that might explain this high level of CI diversity found in C. pipiens populations. Here, we establish that Wolbachia infection is near to or at fixation in worldwide populations of the C. pipiens complex. Wolbachia polymorphism was addressed by sequence analysis of the Tr1 gene, a unique transposable element of the IS5 family, which allowed the identification of five C. pipiens Wolbachia strains, differing either by nucleotide substitution, presence or absence pattern, or insertion site. Sequence analysis also showed that recombination, transposition and superinfection occurred at very low frequencies. Analysis of the geographical distributions of each Wolbachia strain among C. pipiens populations indicated a strong worldwide differentiation independent from mosquito subspecies type, except in the UK. The availability of this polymorphic marker now opens the way to investigate evolution of Wolbachia populations and CI dynamics, in particular in regions where multiple crossing types coexist among C. pipiens populations. PMID- 15813795 TI - Spatial structure of genetic variation and primary succession in the pioneer tree species Antirhea borbonica on La Reunion. AB - In habitats where colonization and extinction are recurrent, the distribution of gene frequencies among patches of suitable habitat may reflect the age structure of different populations. In this study, we quantify population genetic structure for a pioneer tree species, Antirhea borbonica, in a chrono-sequence of primary succession on the lava flows of the Piton de La Fournaise volcano (La Reunion). Using microsatellite loci and amplified fragment length polymorphism (AFLP) markers, we quantified genetic variation within and among populations for early- and late-succession populations in a landscape where extinction and recolonization are recurrent (the 'Grand Brule') and for late-succession populations in a more stable landscape. This study produced three main results. First, we detected no evidence that founder events increase genetic differentiation among colonizing populations; F(ST) values among early- and among late-succession populations were similar. Second, we found no evidence for isolation by distance; genetic distance was not correlated with spatial distance within and among populations. Third, F(IS) values are consistently high in all populations, despite the fact that A. borbonica populations are functionally close to dioecy and thus expected to have an outcrossing mating system. Multiple colonization events from different sources may limit differentiation among young populations and spatial isolation may enhance differentiation among late succession populations. Ecological processes acting during colonization may create the conditions for spatial aggregation within pioneer populations, and thus contribute to the high F(IS) values. PMID- 15813796 TI - Monitoring coyote population dynamics by genotyping faeces. AB - Reliable population estimates are necessary for effective conservation and management, and faecal genotyping has been used successfully to estimate the population size of several elusive mammalian species. Information such as changes in population size over time and survival rates, however, are often more useful for conservation biology than single population estimates. We evaluated the use of faecal genotyping as a tool for monitoring long-term population dynamics, using coyotes (Canis latrans) in the Alaska Range as a case study. We obtained 544 genotypes from 56 coyotes over 3 years (2000-2002). Tissue samples from all 15 radio-collared coyotes in our study area had > or = 1 matching faecal genotypes. We used flexible maximum-likelihood models to study coyote population dynamics, and we tested model performance against radio telemetry data. The staple prey of coyotes, snowshoe hares (Lepus americanus), dramatically declined during this study, and the coyote population declined nearly two-fold with a 1(1/2)-year time lag. Survival rates declined the year after hares crashed but recovered the following year. We conclude that long-term monitoring of elusive species using faecal genotyping is feasible and can provide data that are useful for wildlife conservation and management. We highlight some drawbacks of standard open-population models, such as low precision and the requirement of discrete sampling intervals, and we suggest that the development of open models designed for continuously collected data would enhance the utility of faecal genotyping as a monitoring tool. PMID- 15813797 TI - Facts, faeces and setting standards for the study of MHC genes using noninvasive samples. PMID- 15813799 TI - In search of a new paradigm: mechanisms of sensitization and elicitation of food allergy. AB - In this review of research priorities, presented for the European Commission, food allergy is established as a disease with a considerable impact on modern society. Research paradigms have changed from establishing basic symptomatology and diagnostic methods over allergen characterization to a risk-identification approach looking at which patients are at risk of developing reactions to a certain dose of allergenic food. In order to solve some of the apparent paradoxes of regional, temporal, and species-related differences in sensitization and food allergic reactions, it is suggested to study the basic underlying mechanisms in the cross-field between immunology and gastroenterology. Research priorities should include the molecular basis of cross-reactivity, the digestion and maintenance of antigenicity, the genetics of food allergy, and the immune response to food antigens and effects on the gastrointestinal immune system by food matrices and the microbial gut-flora. Necessary methods with be spanning from molecular biology to large-scale clinical and epidemiological studies. PMID- 15813800 TI - Risks of allergic reactions to biotech proteins in foods: perception and reality. AB - In recent years, significant attention has been paid to the use of biotechnology to improve the quality and quantity of the food supply due in part to the projected growth in the world population, plus limited options available for increasing the amount of land under cultivation. Alterations in the food supply induced by classical breeding and selection methods typically involve the movement of large portions of genomic DNA between different plant varieties to obtain the desired trait. This is in contrast to techniques of genetic engineering which allows the selection and transfers specific genes from one species to another. The primary allergy risk to consumers from genetically modified crops may be placed into one of three categories. The first represents the highest risk to the allergic consumer is the transfer of known allergen or cross-reacting allergen into a food crop. The second category, representing an intermediate risk to the consumer, is the potential for replacing the endogenous allergenicity of a genetically-modified crop. The last category involves expression of novel proteins that may become allergens in man and generally represents a relatively low risk to the consumer, although this possibility has received attention of late. In order to mitigate the three categories of potential allergy risk associated with biotech crops, all genes introduced into food crops undergo a series of tests designed to determine if the biotech protein exhibits properties of known food allergens. The result of this risk assessment process to date is that no biotech proteins in foods have been documented to cause allergic reactions. These results indicate that the current assessment process is robust, although as science of allergy and allergens evolves, new information and new technology should help further the assessment process for potential allergenicity. PMID- 15813801 TI - Pulmonary sarcoidosis. AB - Sarcoidosis is a multisystemic disease of unknown aetiology characterized by the formation of immune granulomas in involved organs. It is a worldwide disease that mainly affects 25-40 years old people with a lifetime incidence rate of 0.85 2.4%. Multiple clinical phenotypes are observed according to presentation, involved organs, disease duration and severity. Sarcoidosis primarily affects the lungs and the lymphatic system. The prevailing pathogenic hypothesis is that various antigens could promote sarcoidosis in genetically susceptible hosts, both these factors modulating the incidence and the clinical phenotype of sarcoidosis. So far, environmental agents have been suspected, including possible mycobacteria and propionibacteria. Interferon-gamma, tumour necrosis factor (TNF)-alpha, interleukin (IL)-12 and IL-18 play a critical role in driving the Th1 commitment in the course of granulomatous process. Evolution of sarcoidosis is often marked by spontaneous resolution within 12-36 months, but can be severe because of chronic cases with pulmonary fibrosis or involving other organs, including heart, central nervous system and eyes. Mortality, ranging between 0.5 and 5%, is most often related to pulmonary fibrosis. Corticosteroids can reverse the granulomatous process, but are only suspensive, and their long-term benefit remains under question. Corticosteroids are recommended when sarcoidosis shows unfavourable clinical tolerance and evolution. Alternative and corticosteroid sparing therapies are of increased interest in difficult cases, while targeted new drugs such as anti-TNF-alpha are still under investigation. PMID- 15813802 TI - EAACI position paper on rhinosinusitis and nasal polyps executive summary. PMID- 15813803 TI - Double-blind placebo-controlled challenges for peanut allergy the efficiency of blinding procedures and the allergenic activity of peanut availability in the recipes. AB - BACKGROUND: A firm diagnosis of double-blind placebo-controlled food challenge (DBPCFC) would facilitate the diagnosis in patients with uncertain history of reaction. Guidelines are lacking for an upper provoking dose and how to hide high concentrations of peanuts. AIM: To develop and evaluate a double-blind recipe with minimum 10% of peanut. To compare the recipe with published recipes regarding blindness, taste, texture and immunoglobulin (Ig)E antibody binding to peanut. METHODS: A recipe (I) with 10% of peanut was developed evaluated and used in DBPCFC. The challenges were followed by development of a concentrated recipe (II) (15% peanut, 25% fat). Recipe II was compared with the only published recipe (III) (11% peanut, 7% fat) regarding taste, texture and availability of peanut. Recipe IV (12% peanut, 10% fat) was developed using the same methods. The binding of IgE in the recipes was measured using an inhibition method. RESULTS: During challenges, one patient reacted after 4 g, emphasizing the need for blinding recipes containing high doses of peanut. Evaluation between recipes II and III, only recipe II was regarded as blind by the taste panels. A tenfold lower availability of peanut protein in the recipe II was found at 50% of inhibition. Recipe IV had a better IgE binding that did not differ from the original peanut extract. CONCLUSION: The peanut taste and texture can be hidden in a challenge medium. The fat content was important for the availability of the allergenic protein in challenges. The availability of allergens must be taken into consideration when used for DBPCFC. PMID- 15813804 TI - Early effects of Asthma Prevention Program on asthma diagnosis and hospitalization in urban population of Poland. AB - BACKGROUND: Asthma is one of the commonest public health problems in Poland and the commonest chronic disorder in children. Lodz Regional Health Insurance Fund was a sponsor of the Prevention Asthma Program in 2000-03, directed at increasing early detection and providing optimal treatment of allergies by specialists in children. METHODS: All funds were divided between 127 primary and 12 specialized care centres participated in the programme. Primary care centre goals were: anamnestic information, mapping of allergy-causing factors, repeated auscultation of the lungs, bronchial dilation test, peak expiratory flow (PEF) measurement at the clinic and at home for 2 weeks. After preliminary diagnosis patients have been send to specialized centres. Specialized care centre goals were as follows: skin testing, spirometry, repeated bronchial dilation test, determination of eosinophilic white blood cells and eosinophil cationic protein (ECP) in blood. RESULTS: Increasing trend of new asthma diagnosis, expressed per 1000 inhabitants, was observed from 0.99 in 2000 to 2.19 in 2003. In the first year of Asthma Prevention Program, we observed more hospital episodes because of asthma exacerbation in comparison with year 1999 but from the second year of programme we showed systematically decreasing number of hospital episodes as a result of asthma exacerbation from 1.48 in 2000 to 0.84 in 2003. We found significantly decreasing trend in duration of hospitalization due to asthma exacerbation (P = 0.001). CONCLUSIONS: Effects of this programmes are: early identification of allergic diseases, mainly asthma, reduced number and shorter duration of hospitalization because of asthma exacerbation and establishing new Asthma Schools Education. PMID- 15813805 TI - Bacterial and fungal components in house dust of farm children, Rudolf Steiner school children and reference children--the PARSIFAL Study. AB - BACKGROUND: Growing up on a farm and an anthroposophic lifestyle are associated with a lower prevalence of allergic diseases in childhood. It has been suggested that the enhanced exposure to endotoxin is an important protective factor of farm environments. Little is known about exposure to other microbial components on farms and exposure in anthroposophic families. OBJECTIVE: To assess the levels and determinants of bacterial endotoxin, mould beta(1,3)-glucans and fungal extracellular polysaccharides (EPS) in house dust of farm children, Steiner school children and reference children. METHODS: Mattress and living room dust was collected in the homes of 229 farm children, 122 Steiner children and 60 and 67 of their respective reference children in five European countries. Stable dust was collected as well. All samples were analysed in one central laboratory. Determinants were assessed by questionnaire. RESULTS: Levels of endotoxin, EPS and glucans per gram of house dust in farm homes were 1.2- to 3.2-fold higher than levels in reference homes. For Steiner children, 1.1- to 1.6-fold higher levels were observed compared with their reference children. These differences were consistently found across countries, although mean levels varied considerably. Differences between groups and between countries were also significant after adjustment for home and family characteristics. CONCLUSION: Farm children are not only consistently exposed to higher levels of endotoxin, but also to higher levels of mould components. Steiner school children may also be exposed to higher levels of microbial agents, but differences with reference children are much less pronounced than for farm children. Further analyses are, however, required to assess the association between exposure to these various microbial agents and allergic and airway diseases in the PARSIFAL population. PMID- 15813806 TI - Early childhood environment related to microbial exposure and the occurrence of atopic disease at school age. AB - BACKGROUND: There is a growing body of evidence that the early childhood environment with respect to day care attendance, older siblings, pet ownership, and early life airway infections may protect from developing atopic disease. Few studies have distinguished between atopic sensitization and symptoms, and none have evaluated independent contributions for all of these different environmental conditions. OBJECTIVE: Examine independent effects on atopic sensitization and symptoms of day care attendance, older siblings, pet ownership, and early infancy's airway disease. METHODS: A cross-sectional survey among 8-13-year-old school children with complete data for 1555 children. RESULTS: After adjustment for confounders, atopic sensitization occurred less frequently in children that had attended a day care centre (OR: 0.73, 95% CI: 0.55-0.98) or had a cat or dog before 2 years of age (OR: 0.78, 95% CI: 0.61-0.99). Having older siblings yielded a nonsignificant trend towards protection (OR: 0.88, 95% CI: 0.70-1.11). For symptoms, there was no relation with having older sibs, day care attendance and pet ownership, although there was a trend towards protection for the combination of atopy and symptoms. In contrast, children with doctors' treated airway disease before age 2, more frequently reported recent symptoms of wheeze, asthma, rhinitis, or dermatitis (all P < 0.05). CONCLUSION: Early life environmental exposure to day care, or pets may protect against atopic sensitization. Protection against symptoms only occurred if atopic sensitization was present as well. PMID- 15813807 TI - The inverse association of salmonellosis in infancy with allergic rhinoconjunctivitis and asthma at school-age: a longitudinal study. AB - BACKGROUND: Respiratory allergies are inversely related to early acquisition of food-borne and fecal-oral infections, consumption of unpasteurized milk, early exposure to stables and high endotoxin concentrations in a farming environment. We tested therefore if infection by Salmonella in early life can protect from development of respiratory allergies later in life. METHODS: During 2003, we studied two groups of Sardinian children (age 6-18 years) who had been hospitalized before 4 years of age (during 1989-2001) with non-typhoid salmonellosis (n = 148) or acute enteritis of nonbacterial etiology (NB enteritis) (n = 167). Allergic rhinoconjunctivitis (AR) and asthma were evaluated by telephonic interview with a ISAAC questionnaire; participants reporting AR and/or asthma were further examined through a complete diagnostic work-up to objectively confirm or exclude current disease. Kaplan-Meier curves and Cox proportional hazard models were used to analyze the role of different types of enteritis on the risk of developing allergic rhinoconjunctivitis or asthma over time. RESULTS: Children who had been hospitalized with salmonellosis had a lower prevalence of allergic rhinoconjunctivitis (eight of 148, 5.4%vs 23 of 167, 13.8%; P = 0.019) or asthma (five of 148, 3.4% vs 21 of 167, 12.6%; P = 0.006) than those who had been hospitalized with NB-enteritis. The proportional hazard of salmonellosis for asthma was 0.23 (95% CI: 0.08-0.67; P < 0.01) and for allergic rhinoconjunctivitis was 0.40 (95% CI: 0.17-0.95; P = 0.04), after adjusting for confounders. DISCUSSION: The strength of the observed associations suggests that Salmonella may contribute to shape the natural history of respiratory allergies. However, further studies are needed to test in other settings the association observed in Sardinian children. We speculate that clinical or subclinical infection by Salmonella may contribute to the atopy protective influence of a traditional farming environment or of areas endemic for food-borne and fecal-oral infections. Food hygiene and prevention of salmonellosis must remain however a public health priority. PMID- 15813808 TI - Association of stem cell factor expression in nasal polyp epithelial cells with aspirin sensitivity and asthma. AB - Mast cells constitute a significant proportion of cells infiltrating nasal polyp tissue, and epithelial cells may release stem cell factor (SCF), a cytokine with chemotactic and survival activity for mast cells. We aimed to assess the expression of SCF in human nasal polyp epithelial cells (NPECs) as related to patients' clinical phenotypes. Nasal polyp tissues were obtained from 29 patients [including nine with aspirin (ASA)-hypersensitivity and 12 with bronchial asthma] undergoing polypectomy for nasal obstruction. Epithelial cells were obtained following 6-week culture of nasal polyps explants. The SCF released into the culture supernatant was assessed by enzyme-linked immunosorbent assay (ELISA) and total SCF mRNA in the polyp tissue was determined by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). For the whole group of patients, the number of polypectomies correlated with expression of SCF mRNA (r = 0.62; P < 0.005), SCF protein in the NPECs supernatants (r = 0.39; P < 0.05) and with density of mast cells in epithelial layer (r = 0.37; P < 0.05) and stromal layer (r = 0.5; P < 0.01) of nasal polyps. The SCF/beta-actin mRNA ratios were significantly higher in ASA-hypersensitive (AH) asthmatics (median 0.97, range: 0.8-1.5) when compared with ASA-tolerant (AT) patients (median 0.5, range: 0.1 0.7; P < 0.001). The SCF protein concentration in NPEC supernatants was also significantly higher in AH asthmatics (median 1.10 pg/microg DNA, range: 0.4-1.9) when compared with AT patients (median 0.1 pg/microg DNA, range: 0.02-1.2; P < 0.001). In the subpopulation of ASA-sensitive asthmatics the number of polypectomies correlated also with the density of mast cells and eosinophils in the polyp tissue. PMID- 15813809 TI - Low levels of CC16 in nasal fluid of children with birch pollen-induced rhinitis. AB - BACKGROUND: Clara cell protein 16 (CC16; secretoglobin 1A1) is an anti inflammatory protein mainly expressed in the epithelial cells in the airways. OBJECTIVE: To compare the levels of CC16 in nasal lavage (NAL) from children with intermittent allergic rhinitis and healthy controls and to study the effect of a local steroid. METHODS: Thirty schoolchildren with birch pollen allergy and 30 healthy controls from the same schools were included in the study. The NAL fluid was collected before the season, during the birch pollen season and, for the patients, after 1 week of treatment with a local steroid. Symptom scores were obtained on every occasion. CC16 and eosinophil cationic protein (ECP) were analyzed with enzyme-linked immunosorbent assay. RESULTS: The nasal fluid levels of CC16 were significantly lower in patients than in controls, before and during pollen season. Before the season, the median CC16 concentrations were 9.1 (range 1.1-117) microg/l in patients and 25.7 (6.1-110.2) microg/l in controls. During the season, the median CC16 concentrations in nasal fluid were 12.9 (2.3-89.7) microg/l in the allergic children and 22.0 (9.5-90.1) microg/l in the healthy controls (P = 0.0005). Symptom scores, nasal fluid eosinophils and ECP were higher in patients during the season. Treatment with a local steroid did not change the CC16 levels. CONCLUSIONS: Nasal fluid CC16 levels were lower in children with birch pollen-induced allergic rhinitis than in healthy controls both before and during the pollen season. We speculate that reduction in anti inflammatory activity by CC16 may contribute to the pathogenesis of allergic rhinitis. PMID- 15813810 TI - Comparison of nasal immunohistology in patients with seasonal rhinoconjunctivitis treated with topical steroids or specific allergen immunotherapy. AB - BACKGROUND: Specific allergen immunotherapy (SIT) and nasal steroids (NS) are considered effective anti-inflammatory treatments for allergic rhinitis, although their mechanism of action differs. OBJECTIVE: The aim of this study was to examine the effect of treatment with NS and SIT on different populations of inflammatory cells in the nasal mucosa and to compare cell numbers before and during the birch pollen season in patients with seasonal allergic rhinitis. METHODS: In a randomized, double-blind, double dummy comparative study, 41 patients with seasonal rhinoconjunctivitis were treated with birch SIT or NS (budesonide 400 microg daily). Treatment with NS started before the birch pollen season and at the same time SIT-treated patients reached the maintenance dose. Nasal biopsies for immunohistochemistry were obtained before the season and start of the treatments and at the peak of the pollen season during treatment. RESULTS: Symptoms of rhinoconjunctivitis increased significantly in both groups during the pollen season but less in the NS-treated group and the difference between the treatment groups was significant at the end of the season (P = 0.03). Immunohistochemistry of nasal biopsies from NS-treated patients showed significantly fewer CD1a+, IgE+ and Fc epsilonRI+ cells during the season compared with preseason (P = 0.02, P = 0.001 and P = 0.0004, respectively) and with seasonal values of the SIT-treated group (P = 0.002, P = 0.002 and P = 0.0004 respectively). CONCLUSION: Treatment with NS but not SIT decreased the numbers of CD1a+, IgE+ and Fc epsilonRI+ cells during the birch pollen season. Our data indicate that treatment with NS has a broader anti-inflammatory range than SIT. PMID- 15813811 TI - Quantitative analysis of IgE antibodies to food and inhalant allergens in 4-year old children reflects their likelihood of allergic disease. AB - BACKGROUND: It is well established that early diagnosis of allergic disease is warranted. METHODS: In a prospective birth cohort study (BAMSE) 3743 children at 4 years of age were included. Children were classified as having any allergic disease, e.g. asthma, suspected allergic rhinitis (suspAR), eczema or oro-gastro intestinal symptoms with questionnaire. Blood was obtained from 2612 of these children and analysed for IgE antibodies (ab) towards 14 common food and airborne allergens. RESULTS: Positive IgE ab results were found in 38% of the children with any allergic disease, whereas such IgE ab results were found in 17% among those without any allergic disease. Furthermore, among children with any allergic disease the median summated IgE ab levels were 10.7 kU(A)/l compared with 1.5 kU(A)/l among those without such symptoms. The highest IgE ab levels were found to birch, peanut, cat and horse. When the sum of the IgE-ab levels towards the selected allergens was at least 34 kU(A)/l, or, alternatively, more than four allergen tests were positive, there was a 75% likelihood of identifying the individual with any allergic disease. To identify those with asthma, as well as those with suspAR, a significant interaction was found for the combination of the sum of IgE-ab levels and number of allergens positive at test. For eczema only, the number of positive allergens at test was associated to the likelihood of such disease. CONCLUSIONS: In children, 4 years of age, allergic disease was frequently not associated with the presence of single positive IgE antibody results, whereas increased IgE ab levels were significantly more prevalent among those with allergic disease. Thus, testing a certain profile of airborne and food allergens, and utilizing the sum of the IgE-ab levels in combination with the number of allergens positive at tests, may represent a more efficient diagnostic tool then to use just single positive IgE-ab results. PMID- 15813812 TI - Peanut- and cow's milk-specific IgE, Th2 cells and local anaphylactic reaction are induced in Balb/c mice orally sensitized with cholera toxin. AB - BACKGROUND: The development of animal models developing specific immunoglobulin (Ig)E presenting the same specificity as human IgE and similar clinical symptoms as those observed in allergic patients are of great interest for the understanding of mechanisms involved in the induction and regulation of food allergy. METHODS: Balb/c female mice were sensitized with whole peanut protein extract (WPPE) by means of intraperitoneal (i.p.) injections with alum or gavages with cholera toxin (CT). The WPPE specific IgE, IgG1 and IgG2a were monitored. Th2 cells activation was analysed assaying interleukin (IL)-4 and IL-5 vs IFNgamma on reactivated splenocytes. Local anaphylactic reaction was evaluated by assaying histamine in faecal samples. The oral sensitization protocol was further extended to cow's milk proteins (CMP). RESULTS: Balb/c mice developed high peanut specific IgE and IgG1 responses either after i.p. or oral sensitizations. In both cases, antibodies were specific to polymer of glycinin fragments, containing polypeptides from Ara h3/4, and to a lesser extent to Ara h1 and Ara h2. Interleukin-4 and IL-5 production were evidenced. Balb/c mice could also be sensitized to CMP, as demonstrated by CMP-specific IL-4 and IL-5 secretions and induction of IgE specific for whole caseins, beta-lactoglobulin, serum bovine albumin and lactoferrin. Of interest was the occurrence of a local anaphylactic reaction in the peanut and CM models. CONCLUSIONS: In contrast with previous authors, Balb/c mice were sensitized and evidenced an allergic reaction after oral administrations of peanut or CMP plus CT, providing an interesting model for further studies on immunopathogenic mechanisms. PMID- 15813813 TI - Nonimmediate reactions to systemic corticosteroids suggest an immunological mechanism. AB - BACKGROUND: Administration of corticosteroids (CS) by different routes may cause varying types of allergic reactions, thereby hampering their further use in affected patients. In order to verify an immunological involvement we evaluated a group of patients with symptoms compatible with nonimmediate allergic reactions to CS. METHODS: Studies included patch and intradermal tests, immunohistochemical studies and controlled administration to reproduce the response. The cytokines interleukin (IL)-4, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha were quantified in peripheral blood during the response. RESULTS: Of 32 subjects evaluated presenting nonimmediate urticaria or exanthema, 21 were finally considered positive after re-exposure. The drugs most frequently involved were betamethasone and dexamethasone. Fewer than half the patients responded to prednisolone whilst some responded to three or more CS. Hydrocortisone and deflazacort were well tolerated by all the patients. Subjects with a positive intradermal or patch test had a perivascular mononuclear cell infiltrate with the presence of CD4 and CD8 lymphocytes positive for CD45RO+ (memory) and CD69 (activation marker) cells. Monitoring peripheral blood during the acute response showed expression of IFN-gamma and TNF-alpha, with downregulation of IL-4. CONCLUSION: Adverse systemic responses to different CS are suggestive of a nonimmediate reaction. The symptoms elicited together with the immunlogical studies suggest a T-cell mediated response. The response to closely related CS was especially marked between betamethasone and dexamethasone, whereas hydrocortisone and deflazacort were well tolerated. PMID- 15813814 TI - A differing pattern of association between dietary fish and allergen-specific subgroups of atopy. AB - BACKGROUND: We examined the role of fish intake in the development of atopic disease with particular reference to the possibility of differential effects on allergen-specific subgroups of sensitization. METHODS: The exposure of interest was parental report of fish intake by children aged 8 years at the 1997 Childhood Allergy and Respiratory Health Study (n = 499). The outcomes of interest were subgroups of atopy: house dust mite (HDM)-pure sensitization [a positive skin prick test (SPT) > or = 2 mm to Der p or Der f only], ryegrass-pure sensitization (a positive SPT > or = 2 mm to ryegrass only); asthma and hay fever by allergen specific sensitization. RESULTS: A significant association between fish intake and ryegrass-pure [adjusted odds ratio (AOR) 0.37 (0.15-0.90)] but not HDM-pure sensitization [AOR 0.87 (0.36-2.13)] was found. Fish consumption significantly decreased the risk for ryegrass-pure sensitization in comparison with HDM-pure sensitization [AOR 0.20 (0.05-0.79)]. CONCLUSIONS: We have demonstrated a differential effect of fish intake for sensitization to different aeroallergens. This may be due to the different timing of allergen exposure during early life. Further investigation of the causes of atopic disease should take into account allergen-specific subgroups. PMID- 15813815 TI - Monophosphoryl lipid A (MPL) promotes allergen-induced immune deviation in favour of Th1 responses. AB - BACKGROUND: Monophosphoryl lipid A (MPL) is a nontoxic derivative of the lipopolysaccharide (LPS) of Salmonella minnesota R595. MPL has been used as an adjuvant in grass and tree pollen vaccines for the treatment of seasonal allergic rhinitis. Little is known about the influence of MPL on cellular responses to allergens in man. We therefore studied the effects of MPL in vitro on peripheral blood mononuclear cells (PBMC) obtained from patients with grass pollen hay fever. METHODS: The PBMCs from 13 subjects were cultured with grass pollen Phleum pratense extract (0, 2 and 20 microg/ml) and MPL (0 and 10 microg/ml; defined as an optimal concentration in preliminary studies) and after 6 days proliferative responses were measured by thymidine incorporation and cytokine production by enzyme-linked immunosorbent assay (ELISA). RESULTS: Proliferative responses were unaffected by the presence of MPL whereas MPL induced a significant increase in allergen-induced interferon (IFN)-gamma production [allergen alone, 645 +/- 466 pg/ml (mean +/- SE) vs allergen + MPL, 3232 +/- 818 pg/ml; P < 0.001]. In addition, there was a significant decrease in interleukin (IL)-5 production (4307 +/- 1030 pg/ml vs 2997 +/- 826 pg/ml; P < 0.01). Although MPL alone could induce modest increases in IL-10 production, MPL did not influence the production of this cytokine in allergen-stimulated cultures. Addition of neutralizing antibody against IL-12 resulted in 95% inhibition of MPL-induced IFN-gamma production. Depletion of monocytes from the culture system abrogated the effects of MPL on elevated cytokine production. CONCLUSIONS: In summary, use of MPL with grass pollen extract results in immune deviation of allergen-induced peripheral Th2 cell responses in favour of 'protective' Th1 responses in an IL-12 and monocyte dependent fashion. PMID- 15813816 TI - Serum thymus and activation-regulated chemokine, macrophage-derived chemokine and eotaxin as markers of severity of atopic dermatitis. AB - BACKGROUND: Expression of CCR4 ligands, such as thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), leads to preferential influx of T-helper (Th) 2-type lymphocytes to the lesional skin in atopic dermatitis (AD). Eotaxin, like the CCR3 ligand, is an important contributor of eosinophils recruitment in the course of AD. These chemokines are assumed to play an important role in the pathomechanism of AD. METHODS: In this study, the serum concentration of TARC, MDC, eotaxin and total immunoglobulin E (IgE) in AD patients and healthy people were compared. Correlation between the studied indices and activity of AD was established. Severity of AD was assessed according to the SCORAD score. The study comprised 44 healthy people and 43 patients with AD. The serum concentrations of TARC, MDC, eotaxin and IgE were measured with the use of enzyme-linked immunosorbent assay kits. RESULTS: The serum levels of TARC, MDC, eotaxin and IgE appeared to be significantly higher in patients with AD than in healthy people. A strong positive correlation was revealed between the levels of TARC, MDC, total IgE in serum of patients with AD and SCORAD. In contrast, no significant relationship was found for the serum eotaxin concentration and TARC, MDC, IgE or disease severity. CONCLUSION: Our findings indicate that TARC and MDC are actively involved in the pathogenesis of AD and their expression, opposite to that of eotaxin, is strongly associated with clinical picture of atopic dermatitis. PMID- 15813817 TI - Possible circadian variation of serum mast cell tryptase concentration. AB - BACKGROUND: A temporarily elevated level of serum mast cell tryptase (ST) indicates mast cell activation and occurs in systemic anaphylactic reactions (SAR). We measured ST following a sting challenge in vespid venom-allergic patients treated with venom immunotherapy (VIT) and in healthy controls, respectively. AIM OF THE STUDY: To assess changes of ST over time in vespid venom allergic patients at the occasion of a re-sting and in healthy controls. METHODS: A sting challenge was performed in 20 patients on vespid VIT to monitor efficacy of VIT. ST was measured between 9.00 and 10.00 a.m. (baseline). Sting challenge was performed at 2.00 p.m., and ST was determined again 20 min, 90 min and 18 h later. Measurements at corresponding times of the day were done in nine healthy controls. RESULTS: One patient developed a mild SAR to the sting challenge which was associated with a temporary increase of ST. In the other 19 patients who tolerated the sting challenge without SAR ST decreased significantly by 18.0% (median, range 8.3-36.7%). Twenty minutes after the sting when compared with baseline levels (P < 0.001), a significant decrease of ST was still present after 90 min (median 13.7%) (P < 0.001), but not after 18 h (P = 0.57). A comparably significant temporary decline was found in controls. CONCLUSIONS: The temporary decline of ST in patients and in controls suggests a circadian variation of ST concentration. A normal diurnal pattern of ST concentration after sting challenge is associated with successful treatment. PMID- 15813818 TI - Anti-IL-5 recombinant humanized monoclonal antibody (mepolizumab) for the treatment of atopic dermatitis. AB - BACKGROUND: Eosinophils may play an important role in the pathogenesis of atopic dermatitis (AD). Interleukin-5 is essential for eosinophil growth, differentiation and migration. A monoclonal antibody to human interleukin-5 (mepolizumab) was developed for atopic diseases. This study was designed to study the effect of mepolizumab in AD. METHODS: Two single doses of 750 mg mepolizumab, given 1 week apart, were studied in patients with moderate to severe AD using a randomized, placebo-controlled parallel group design. The primary endpoint of 'success' to treatment was defined as the percentage of patients with at least 'marked improvement' after 2 weeks as assessed by the Physician's Global Assessment of Improvement (PGA). Furthermore, SCORing AD (SCORAD), pruritus scoring, number of blood eosinophils and serum thymus and activation-regulated chemokine (TARC) values served as secondary endpoints. Fluticasone propionate cream 0.05%, once daily could be used as rescue medication from day 16 if no improvement was recorded. RESULTS: Eighteen patients received mepolizumab and 22 placebo treatment. Peripheral blood eosinophil numbers were significantly reduced in the treatment group compared with placebo (P < 0.05). No clinical success was reached by PGA assessment (P = 0.115), SCORAD (P = 0.293), pruritus scoring and TARC values in the mepolizumab-treated group compared with placebo. However, modest improvement (<50% improvement) assessed by PGA was scored significantly more in the mepolizumab-treated group compared with placebo (P < 0.05). CONCLUSION: Two single doses of 750 mg mepolizumab did not result in clinical success in patients with AD, despite a significant decrease in peripheral blood eosinophils. PMID- 15813819 TI - Predictors of indoor exposure to mouse allergen in urban and suburban homes in Boston. AB - BACKGROUND: Mouse allergen exposure is prevalent among urban children with asthma. Little is known about mouse allergen exposure in children at risk for the development of allergic diseases. AIMS OF THE STUDY: To assess indoor mouse allergen exposure in early life among children with parental history of asthma or allergies. METHODS: Prospective birth cohort study of 498 children with a history of allergy or asthma in at least one parent living in metropolitan Boston. RESULTS: Of the 498 participating children, 357 (71.7%) resided outside the city of Boston and 439 (90.7%) lived in households with incomes > 30,000 dollars. Mouse allergen was detected in 42% of the homes of study participants. In a multivariate analysis adjusting for sex, income, and endotoxin, black race [odds ratio (OR) = 3.0; 95% confidence interval (CI) = 1.3-6.6, P = 0.009], signs of mice in the home at age 2-3 months (OR = 3.0; 95% CI = 1.6-5.6, P = 0.0006), and kitchen cockroach allergen levels > or = 0.05 to < 2 U/g (OR = 1.8; 95% CI = 1.1 3.2, P = 0.02) were associated with detectable mouse allergen in the kitchen. In this model, living in a single detached house was inversely associated with detectable kitchen mouse allergen levels (OR = 0.4; 95% CI = 0.2-0.6, P = 0.0001). CONCLUSION: Infants with a parental history of asthma or allergies are commonly exposed to mouse allergen in their homes. Among infants at high risk for atopy, predictors of increased mouse allergen levels included black race, reported mice exposure, and moderate levels of cockroach allergen. PMID- 15813820 TI - Bacillus clausii effects in children with allergic rhinitis. PMID- 15813821 TI - Flow-assisted diagnosis of anaphylaxis to patent blue. PMID- 15813822 TI - Anaphylaxis to macrogol 4000 after a parenteral corticoid injection. PMID- 15813823 TI - Urticaria due to pentoxyfylline. PMID- 15813824 TI - An unusual pattern of meat allergy. PMID- 15813825 TI - The Child Behaviour Checklist in the study of emotional and behavioural problems in children with asthma. PMID- 15813826 TI - Safety of celecoxib in patients with adverse reactions to nimesulide. PMID- 15813827 TI - No rHev b 12-specific IgE-response in children sensitized to natural rubber latex. PMID- 15813828 TI - Review article: smoking cessation as primary therapy to modify the course of Crohn's disease. AB - This article aims to offer an updated review of the effects of smoking on inflammatory bowel disease, and provide a review of the methods of achieving smoking cessation. A systematic review of Embase and Medline databases was conducted. Smoking causes opposing effects on ulcerative colitis and Crohn's disease. The odds ratio of developing ulcerative colitis for smokers compared with lifetime non-smokers is 0.41. Conversely, smokers with Crohn's disease have a more aggressive disease requiring more therapeutic intervention. Smoking cessation is associated with a 65% reduction in the risk of a relapse as compared with continued smokers, a similar magnitude to that obtained with immunosuppressive therapy. Although difficult to achieve smoking cessation can best be encouraged by accessing appropriate counselling services, nicotine replacement therapy and bupropion. Using a combination of these treatments there is an improved chance of success of up to 20% compared with an unassisted quit attempt. Smoking cessation unequivocally improves the course of Crohn's disease and should be a primary therapeutic aim in smokers with Crohn's disease. PMID- 15813829 TI - Review article: current management of primary sclerosing cholangitis. AB - The management of primary sclerosing cholangitis (PSC) is hindered by incomplete understanding of the pathogenesis of the disease and the lack of good prognostic models. Few large randomized controlled trials of drug therapy have been published. Best practice in the management of PSC is currently based therefore on careful interpretation of the available evidence, close observation of individual patients and clinical experience of the disease. Drug therapy is useful for alleviating symptoms. Ursodeoxycholic acid may slow progression of the disease and reduce the frequency of complications. Consensus is emerging on the issues of screening for the malignant complications of PSC and the indications for liver transplantation are becoming broader and encompassing the earliest stages of cholangiocarcinoma. In view of the rarity of the disease in the general population, large international collaborations to study PSC are necessary to provide clearer answers in areas of uncertainty, and these are now beginning to emerge. PMID- 15813830 TI - Review article: current management of renal dysfunction in the cirrhotic patient. AB - The United Network for Organ Sharing database revealed that over the last 4-5 years, an average of 1800 patients were removed from the cadaveric waiting list annually because of patients' death and an additional 400-500 were removed from the list because of the severity of their illnesses. The pre-transplant evaluation process, therefore, requires careful and continued assessment of the patient's pulmonary, cardiac and renal function among others. This article describes a systematic approach to the evaluation and management of renal dysfunction complicating the course of advanced liver disease, the pathogenic mechanisms and current recommendations for the treatment of hepatorenal syndrome, and the indications for combined liver-kidney transplantation. PMID- 15813831 TI - Intragastric acidity during treatment with esomeprazole 40 mg twice daily or pantoprazole 40 mg twice daily--a randomized, two-way crossover study. AB - BACKGROUND: Patients with severe or complicated reflux disease may require higher than standard doses of a proton pump inhibitor for sufficient acid suppression. AIM: To test the hypothesis that esomeprazole 40 mg twice daily is superior to pantoprazole 40 mg twice daily in lowering intragastric acidity. METHODS: In a randomized, single-blinded, two-way crossover study, healthy subjects received esomeprazole 40 mg twice daily or pantoprazole 40 mg twice daily orally for five consecutive days. Continuous ambulatory 24-h intragastric pH was recorded on day 5 of each treatment. RESULTS: Thirty subjects were analysed. Esomeprazole provided significantly higher intragastric pH-values over the 24-h period [median intragastric pH 6.4 for esomeprazole and 5.1 for pantoprazole (P < 0.00005)]. Intragastric pH > 4 was maintained for 21.1 h with esomeprazole and 16.8 h with pantoprazole (P < 0.0001). An intragastric pH > 4 for more than 16 h was achieved in 96.7 and 56.7% of subjects, respectively (P = 0.0002). During night-time the proportion of time with intragastric pH > 4 was 85.4% with esomeprazole and 63.6% with pantoprazole (P = 0.0001). Nocturnal acid break through occurred less frequently on esomeprazole. CONCLUSIONS: Esomeprazole 40 mg twice daily provides better and more consistent intragastric acid control than pantoprazole 40 mg twice daily. PMID- 15813832 TI - Normalization of oesophageal pH does not guarantee control of duodenogastro oesophageal reflux in Barrett's oesophagus. AB - BACKGROUND: Proton-pump inhibitors are effective at preventing the acid component of gastro-oesophageal refluxate from entering the oesophagus. It is not clear whether proton-pump inhibitors prevent duodenogastro-oesophageal reflux. AIM: To measure oesophageal exposure to duodenogastro-oesophageal refluxate while on proton-pump inhibitors in patients with Barrett's oesophagus. METHODS: Twenty five patients (23 male) with Barrett's oesophagus underwent 24 h oesophageal pH and Bilitec 2000 monitoring while on omeprazole 40 mg/day (n = 19) or omeprazole 60 mg/day (n = 6). All patients were undergoing argon plasma ablation of their Barrett's epithelium as part of a clinical trial and the Bilitec measurements were only carried out after the ablation had been completed. RESULTS: 20 of 25 (80%) patients had a normal oesophageal pH profile. Fifteen of the 25 (60%) had abnormal oesophageal exposure to bile as measured by Bilitec 2000. Of the 20 patients who had a normal 24 h oesophageal pH profile, 11 (55%) had pathological exposure to bile in their oesophagus. CONCLUSION: Complete acid suppression does not guarantee elimination of duodenogastro-oesophageal reflux. PMID- 15813833 TI - Long-term acid suppression by omeprazole in gastro-oesophageal reflux disease patients does not lead to anti-gastric autoantibody production. AB - BACKGROUND: Helicobacter pylori-associated atrophy of the gastric corpus is associated with the presence of anti-canalicular autoantibodies. Also, long-term profound acid suppression in H. pylori-infected subjects may cause atrophic corpus gastritis. AIM: To investigate whether long-term acid suppression by omeprazole leads to antigastric autoantibodies. METHODS: Fifty patients, of which 34 H. pylori-positive on entry of the study, were treated with omeprazole (20-40 mg once daily) for reflux oesophagitis, and were evaluated for anti-gastric autoantibody responses by immunohistochemistry before and after treatment. H. pylori was not eradicated and patients were followed for an average of 6.6 years (range 3-14.1 years). In addition to immunohistochemistry, anti-H(+), K(+)-ATPase reactivity was assessed by Western blot in paired sera of 41 patients (26 H. pylori-positive and 15 uninfected) and results are critically evaluated. RESULTS: In immunohistochemistry, all patients were negative for anti-canalicular autoantibodies when omeprazole therapy started, except for two patients with corpus-predominant gastritis in the presence of H. pylori. One patient, who was H. pylori-negative, newly developed an anti-canalicular antibody response during therapy. CONCLUSIONS: Our results indicate that, as compared with non-infected patients, long-term profound acid suppression therapy in H. pylori-infected gastro-oesophageal reflux disease patients does not increase or accelerate gastric autoimmunity. PMID- 15813834 TI - Absorbable vs. non-absorbable antibiotics in the treatment of small intestine bacterial overgrowth in patients with blind-loop syndrome. AB - BACKGROUND: Small intestine bacterial overgrowth is associated with the presence of predisposing conditions, acting through different mechanisms. Therefore, the failure to define a standardized therapy may be due to a methodological bias: to treat a condition characterized by different pathophysiological mechanisms with the same pharmacological approach. Non-absorbable antibiotics could have a lower efficacy than absorbable drugs in patients with blind loops which exclude a portion of the intestine from the transit. AIM: To evaluate the efficacy of absorbable vs. non-absorbable antibiotics in this subgroup of patients. METHODS: A group of small intestine bacterial overgrowth patients with total gastrectomy or gastrojejunostomy and blind loop underwent a therapeutic trial comparing rifaximin to metronidazole. Seven patients underwent a course of rifaximin followed by a course of metronidazole on recurrence of symptoms. To compare the effect of the drugs, another two groups of patients underwent two consecutive courses of rifaximin or metronidazole. Hydrogen breath test after glucose administration and symptom severity measurement were performed. RESULTS: Both drugs reduced breath H(2) excretion but a much better improvement was achieved after metronidazole. Symptom improvement was higher after metronidazole. CONCLUSION: Metronidazole is more effective than rifaximin for the treatment of small intestine bacterial overgrowth associated with the presence of a blind loop. PMID- 15813835 TI - Effect of the administration schedule on the therapeutic efficacy of oral pancreatic enzyme supplements in patients with exocrine pancreatic insufficiency: a randomized, three-way crossover study. AB - BACKGROUND: Oral pancreatic enzyme supplements should be properly administered in order to ensure an adequate gastric mixing with the food and simultaneous gastric emptying with the chyme. AIM: To evaluate, in a prospective, randomized, open, comparative, three-way, crossover study, the effect of the administration schedule on the efficacy of oral pancreatic enzymes for the treatment of exocrine pancreatic insufficiency. METHODS: Twenty-four consecutive chronic pancreatitis patients with maldigestion secondary to exocrine pancreatic insufficiency were treated with 40 000 U lipase in the form of capsules containing enteric-coated mini-microspheres. Capsules were taken just before meals (schedule A), just after meals (schedule B) or distributed along with meals (schedule C) for three consecutive 1-week crossover periods in a randomized order. Fat digestion before and during the three treatment periods was evaluated by an optimized mixed (13)C triglyceride breath test. RESULTS: Before therapy, the (13)CO(2) recovery in the breath test was 23.8 +/- 15.8% (normal >58.0%). During therapy, the (13)CO(2) recovery tended to be higher when capsules were taken along with meals ((13)CO(2) recovery 61.4 +/- 21.4%) or just after meals ((13)CO(2) recovery 60.6 +/- 21.8%) than when taken just before meals ((13)CO(2) recovery 53.9 +/- 20.3%). The percentage of patients who normalized fat digestion under therapy was 50, 54 and 63% with schedules A, B and C respectively. CONCLUSIONS: The efficacy of pancreatic enzyme supplements for the treatment of exocrine pancreatic insufficiency may be optimized by administration during or after meals. PMID- 15813836 TI - Anxiety, depression and psychotropic medication use in patients with persistent upper and lower gastrointestinal symptoms. AB - BACKGROUND: Limited information is available about anxiety, depression and psychotropic medication use in patients referred for gastrointestinal endoscopy. AIM: To determine anxiety and depression and its association with endoscopic findings in a representative sample of patients with persistent gastrointestinal symptoms prior to endoscopy. METHODS: Patients referred to the hospital for endoscopy between February 2002 and February 2004 were asked to score anxiety and depression on the Hospital Anxiety and Depression Scale 2 weeks prior to endoscopy. Information about endoscopic diagnoses was obtained from medical files. RESULTS: A total of 1298 subjects was studied (600 upper gastrointestinal endoscopies and 698 lower gastrointestinal endoscopies). Patients referred for upper gastrointestinal endoscopy used most psychotropic agents (24%; odds ratio = 3.1; 95% confidence interval = 2.3-4.2), especially patients with an organic abnormality when compared with patients without an organic abnormality (42% versus 8%; odds ratio = 8.6; 95% confidence interval = 5.4-14.0). Patients with colonic polyps were more anxious (odds ratio = 1.7; 95% confidence interval = 1.0 2.9) and depressed (odds ratio = 1.8; 95% confidence interval = 1.1-3.1) than other patients referred for lower gastrointestinal endoscopy. CONCLUSIONS: There is no difference in anxiety nor depression between patients with and without organic abnormalities at endoscopy. Patients with colonic polyps are more anxious and depressed than other patients referred for lower gastrointestinal endoscopy. Psychotropic medication use is highest among patients with an organic abnormality in the proximal gastrointestinal tract. PMID- 15813837 TI - Is the management of hepatitis C patients appropriate? A population-based study. AB - BACKGROUND: In order for hepatitis C patients to receive antiviral treatment, they must reach medical care. AIM: To assess the proportion of patients reaching medical care after hepatitis C diagnosis in a general population (1 006 171 inhabitants) in France. METHODS: Between 1994 and 1999, 1508 cases were diagnosed, of which 1251 were eligible for the study. RESULTS: Two-hundred and two patients did not have any medical care; among them, 55.4% had normal alanine transferase, 58.4% had risk factors related to lifestyle and 22.8% were alcoholics. Amongst the 1049 other patients, 41.6% had a liver biopsy, 25.0% were treated. Treatment was more often carried out in males than in females (OR: 1.59; P = 0.001), and in patients under 65 than in older patients (OR: 2.22; P < 0.008). Among non-treatment reasons, alcoholism (P = 0.001), drug-addiction (P = 0.04) and escaping monitoring (P = 0.04) were more frequent in males than in females, whereas normal alanine transferase was more frequent in females than in males (P = 0.004). Amongst 278 patients with a Metavir score >A1F1, 71 (25.5%) did not undergo treatment. CONCLUSION: In a general population, one patient in six did not receive on-going health care; a quarter of patients with a Metavir score >A1F1 did not receive any treatment. These results showed insufficient clinical management, which could compromise the effectiveness of treatment in general population. PMID- 15813838 TI - Mucosa-associated bacteria in ulcerative colitis before and after antibiotic combination therapy. AB - BACKGROUND: We proposed that Fusobacterium varium is one of the causative agents in ulcerative colitis. AIM: To examine the efficacy of antibiotic combination therapy against F. varium and to investigate the mucosa-associated bacteria before and after the therapy using a new molecular approach. METHODS: Twenty patients with ulcerative colitis were randomly assigned into the antibiotic treatment group (amoxicillin, tetracycline and metronidazole for 2 weeks) and no antibiotics group. Clinical assessment, colonoscopic and histological evaluations were performed at 0 and 3-5 months after the treatment. DNA from mucosal bacteria was isolated from biopsy specimens. We investigated the mucosa-associated bacterial components by terminal restriction fragment length polymorphism with the restriction enzyme HhaI and MspI, and quantified the change in the number of bacteria by real-time polymerase chain reaction. Immunohistochemical detection of F. varium in biopsy specimens was also performed. RESULTS: After the treatment, the clinical assessment, colonoscopic and histological scores improved in the antibiotic group compared with the control group. Three peaks of terminal restriction fragment length polymorphism decreased after treatment only in the antibiotic group. Eubacterium rectale, Dorea formicigenerans, Clostridium clostridioforme and F. varium were included in these peaks. Based on the real time polymerase chain reaction study, only F. varium was significantly reduced after treatment. In the immunostaining, post-treatment scores in treatment group were significantly lower than that in control group. CONCLUSIONS: Antibiotics combination therapy was effective for ulcerative colitis. The number of mucosa associated F. varium significantly decreased after the treatment. PMID- 15813839 TI - Controlled trial of interventions to increase testing and treatment for Helicobacter pylori and reduce medication use in patients with chronic acid related symptoms. AB - BACKGROUND: Many symptomatic patients take proton pump inhibitors or histamine-2 blockers for years and those without gastro-oesophageal reflux disease might benefit from Helicobacter pylori eradication. AIM: To increase testing and treatment of H. pylori and reduce chronic use of proton pump inhibitors and histamine-2 blockers. METHODS: We conducted a three-armed controlled trial in 14 managed care practices. We included adults who used proton pump inhibitors or histamine-2 blockers for >1 year and excluded those with gastro-oesophageal reflux disease or previous endoscopy. We compared usual care (n = 312 patients from 6 practices) to low-intensity (n = 147 from 3 practices) and high-intensity (n = 122 from 5 practices) interventions. Low-intensity intervention consisted of guidelines, patient-lists, and a "toolkit"; high-intensity intervention added academic group detailing by a gastroenterologist with reinforcement by pharmacists. RESULTS: Compared with usual care, the high-intensity intervention increased H. pylori test-ordering (29% versus 9% at 12 months, P = 0.02). About half (23 of 58) of patients tested positive and 22 received eradication treatments. The high-intensity intervention decreased proton pump inhibitor use by 9% per year (P = 0.028), but did not alter histamine-2 blocker use. The low intensity intervention was ineffective. CONCLUSIONS: Providing guidelines, patient-lists, and toolkits was no better than usual care. Adding group detailing and pharmacist reinforcements led to improvements in H. pylori management and decreases in proton pump inhibitor use. PMID- 15813840 TI - A controlled trial of ondansetron in the pruritus of cholestasis. AB - BACKGROUND: In patients with pruritus of cholestasis, response to conventional drug treatment may be unsatisfactory. Activation of 5-hydroxytryptamine receptors on dermal sensory nerve-endings plays a role in the perception of pruritus. The 5 hydroxytryptamine(3) receptor antagonist, ondansetron, has been used in the treatment of pruritus of cholestasis, but there are few controlled data. AIM: To determine whether ondansetron is effective in treating the pruritus of cholestasis. METHODS: A total of 19 patients with resistant pruritus were randomized, double blind, to receive either ondansetron 8 mg or placebo as a single intravenous bolus, followed by oral ondansetron 8 mg or placebo twice daily for 5 days. Patients' perception of pruritus was recorded hourly using a visual analogue scale, and scratching activity measured by means of a piezo electric crystal attached to the fingernail. RESULTS: Mean pruritus score using visual analogue scale and scratching activity were reduced on the first treatment day compared with baseline in both the ondansetron and placebo groups (P < 0.05), but there were no significant differences in mean pruritus perception or scratching activity between the two groups. CONCLUSION: Ondansetron was of no benefit in this group of pruritic patients during short-term treatment. PMID- 15813841 TI - Long-term follow-up of Helicobacter pylori eradication therapy in Vietnam: reinfection and clinical outcome. AB - AIM: To assess the long-term Helicobacter pylori reinfection rates, as well as the clinical outcome in peptic ulcer disease patients in Vietnam. METHOD: At a 1 year evaluation of H. pylori eradication treatment in 226 peptic ulcer patients, long-term H. pylori status was assessed with serology and/or culture, peptic ulcer status by gastroscopy, and DNA-fingerprinting performed with random amplified polymorphic DNA and restriction fragment polymorphism. RESULT: Follow up was performed a mean 11 months after the post-treatment evaluation on day 30 after beginning of treatment. The overall reinfection rate was 23.5%, with 58.8% of the strains being identical to the pre-treatment isolates and 41.2% being different. Peptic ulcer was found in 22.9% of the reinfected patients and in 6.3% of the non-reinfected. At the long-term follow-up of successful eradication cases, 89.8% of the patients were free of peptic ulcer disease. The corresponding result was 58.7% in patients in whom H. pylori eradication failed. CONCLUSION: Following successful H. pylori eradication, reinfection with H. pylori in patients in Vietnam was found to be higher than in industrialized countries but the long-term recurrence of peptic ulcer disease was still low. Helicobacter pylori eradication treatment is therefore of value also in developing countries as the rate of peptic ulcer disease was low at the 1-year follow-up. PMID- 15813842 TI - Bone relapse in a patient with acute lymphoblastic leukaemia. PMID- 15813843 TI - Dutcher-Fahey intranuclear inclusions in multiple myeloma. PMID- 15813844 TI - Primary and secondary thrombocytosis in childhood. AB - This review summarizes current data on the pathomechanisms and clinical aspects of primary and secondary thrombocytosis in childhood. Primary thrombocytosis is extremely rare in childhood, mostly diagnosed at the beginning of the second decade of life. As in adults, the criteria of the Polycythemia Vera Group are appropriate to diagnose primary thrombocytosis. The pathomechansims of non familial forms are complex and include spontaneous formation of megakaryopoietic progenitors and increased sensitivity to thrombopoietin (Tpo). Familial forms can be caused by mutations in Tpo or Tpo receptor (c-mpl) genes. These mutations result in overexpression of Tpo, sustained intracellular signalling or disturbed regulation of circulating Tpo. Treatment of primary thrombocytosis is not recommended if platelet counts are <1500/nl and bleeding or thrombosis did not occur in patient's history. In severe cases, decision on treatment should weigh potential risks of treatment options (hydroxyurea, anagrelide) against expected benefits for preventing thrombosis or haemorrhage. Secondary thrombocytosis is frequent in children, in particular in the first decade of life. Hepatic Tpo production is stimulated in acute response reaction to a variety of disorders. Thrombosis prophylaxis is not required, even at platelet counts >1000/nl, except for cases with additional prothrombotic risk factors. PMID- 15813845 TI - Gaucher disease: pathological mechanisms and modern management. AB - Gaucher disease, the most common lysosomal storage disorder, is caused by the defective activity of the lysosomal enzyme, acid-beta-glucosidase (GlcCerase), leading to accumulation of glucosylceramide (GlcCer), particularly in cells of the macrophage lineage. Nearly 200 mutations in GlcCerase have been described, but for the most part, genotype-phenotype correlations are weak, and little is known about the down-stream biochemical changes that occur upon GlcCer accumulation that result in cell and tissue dysfunction. In contrast, the clinical course of Gaucher disease has been well described, and at least one treatment is available, namely enzyme replacement therapy. One other treatment, substrate reduction therapy, has recently been marketed, and others are in early stages of development. This review, after discussing pathological mechanisms, evaluates the advantages and disadvantages of existing therapies. PMID- 15813846 TI - Cellular drug sensitivity in MLL-rearranged childhood acute leukaemia is correlated to partner genes and cell lineage. AB - Rearrangements in the 11q23 region, the site of the mixed lineage leukaemia (MLL) gene, are found in both childhood acute myeloid (AML) and lymphoblastic (ALL) leukaemia. We studied the in vitro drug resistance by the fluorometric microculture cytotoxicity assay (FMCA) in 132 children with AML and 178 children with ALL (aged 0-17 years). In AML, children with t(9;11) (n = 10) were significantly more sensitive to cytarabine (P < 0.001) and doxorubicin (P = 0.005) than non-11q23 rearranged patients (n = 108). Children with other 11q23 rearrangements (n = 14) differed less from non-rearranged children. The 'AML profile' common to all three groups included relative resistance to glucocorticoids and vincristine. In ALL, children with 11q23 rearrangement (n = 22) were significantly more sensitive to cytarabine (P = 0.026) than children without 11q23 rearrangement (n = 156), also after stratification for white blood cell count. In conclusion, the findings indicate that the cellular drug resistance is correlated to both the cell lineage and the type of 11q23 rearrangement. High cellular sensitivity to cytarabine and doxorubicin might explain the excellent treatment results in children with AML and t(9;11). The present study supports the strategy of contemporary protocols to include high dose cytarabine in the treatment of 11q23-positive patients both in AML and ALL. PMID- 15813847 TI - Hodgkin's disease variant of Richter's syndrome in chronic lymphocytic leukaemia patients previously treated with fludarabine. AB - The transformation of chronic lymphocytic leukaemia (CLL) into large-cell lymphoma (Richter's syndrome, RS) is a well-documented phenomenon. Only rarely does CLL transform into Hodgkin's lymphoma (HL). To further analyse the clinico pathological and genetic findings in the HL variant of RS, we performed a single institution study in four patients, who developed HL within a mean of 107 months after diagnosis of CLL. All were treated with fludarabine. Three cases were Epstein-Barr virus (EBV)-associated mixed cellularity (MC) HL, the fourth was nodular sclerosis (NS) HL without EBV association. The sites involved by HL included supra- and infradiaphragmal lymph nodes and the tonsils; stage IV disease was also documented. All patients presented with CLL treatment-resistant lymphadenopathies and B-symptoms. In two of the MC cases, molecular analysis performed on CLL samples and microdissected Hodgkin and Reed-Sternberg cells (HRSC) suggested a clonal relationship, while in NS no indication of a clonal relationship was detected. In summary, HL can occur in CLL patients at any site, up to 17 years after initial diagnosis, especially after treatment with fludarabine. The majority present with B-symptoms and CLL treatment-resistant lymphadenopathy, are of the MC type, clonally related to CLL and might be triggered by an EBV infection. PMID- 15813848 TI - Impact of epoetin-beta on survival of patients with lymphoproliferative malignancies: long-term follow up of a large randomized study. AB - Recent studies have suggested that epoetin treatment of anaemia may influence the survival of patients with cancer. We conducted an analysis of long-term survival in patients with lymphoproliferative malignancies treated with epoetin-beta or placebo in a large-scale study. This was a randomized, double-blind trial in which patients with transfusion-dependent anaemia and lymphoproliferative malignancy received epoetin-beta 150 IU/kg or placebo three times weekly for 16 weeks. Long-term survival data were analysed by standard Kaplan-Meier methods and differences between groups were assessed using a log-rank test. The intention-to treat population consisted of 343 patients (epoetin-beta, n = 170; placebo, n = 173). There were no major differences between the two treatment groups in demographic or clinical characteristics/prognostic factors. A total of 110 (65%) patients died in the epoetin-beta group (censored, n = 60) and 109 (63%) died in the placebo group (censored, n = 64) up to the end of long-term follow up. Kaplan Meier curves for survival were similar in both groups. Median survival was 17 months with epoetin-beta and 18 months with placebo. A log-rank test indicated no significant difference in survival (P = 0.76). This long-term follow up indicated that epoetin-beta has no significant effect on survival compared to placebo in anaemic patients with lymphoproliferative malignancies. PMID- 15813849 TI - Del (9q) AML: clinical and cytological characteristics and prognostic implications. AB - Del (9q) is a recurrent cytogenetic abnormality in acute myeloid leukaemia (AML). We report an analysis of 81 patients with del(9q) as a diagnostic karyotypic abnormality entered into the Medical Research Council AML trials 10, 11 and 12. Patients were divided into three groups: (i) Sole del (9q), 21 patients; (ii) Del(9q) in association with t(8;21), 29 patients; (iii) Del(9q) in association with other cytogenetic abnormalities, 31 patients. Sole del(9q) was associated with a characteristic bone marrow phenotype at diagnosis: a single Auer rod was found in all cases examined. There was also an association with erythroid dysplasia (74%) and granylocytic lineage vacuolation (90%). The incidence of all three of these features was significantly higher (P < 0.05) in the sole del(9q) group compared with control cases lacking del(9q). The overall survival (OS) of all 81 patients was compared with a control group of 1738 patients with normal cytogenetics entered in the same trials over the period of investigation. The 5 year OS for patients with del(9q) was 45%, compared with 35% for the control group (P = 0.09). Patients with del(9q) in association with t(8;21) had a 5-year OS of 75%, which was significantly better than the groups with either sole del(9q) (40%) and del(9q) with other abnormalities (26%; P = 0.008). Karyotyping indicated a common area of deletion in the region 9q21-22, which was present in 94% of cases. It is likely that the deletion of single or multiple tumour suppressor genes located in this region may underlie the pathogenesis of del (9q) AML. PMID- 15813850 TI - A UGT2B17-positive donor is a risk factor for higher transplant-related mortality and lower survival after bone marrow transplantation. AB - We recently identified a human minor histocompatibility (H) antigen, encoded by UDP glycosyltransferase 2 family, polypeptide B17 (UGT2B17), whose immunogenicity results from differential expression in donor and recipient cells as a consequence of a homozygous deletion of the UGT2B17 gene. UGT2B17 is highly expressed in the liver and colon, which are major targets for graft-versus-host disease (GVHD). To assess the significance of homozygous UGT2B17 gene deletion in allogeneic haematopoietic stem cell transplantation (HSCT), we analysed DNA from 435 stem cell transplant recipients with a haematological malignancy and their human leucocyte antigen-identical unrelated bone marrow donors using sequence specific primer polymerase chain reaction. Homozygous deletion of the UGT2B17 gene was observed in 85% of normal donors and in 82% of patients. The analysis showed no significant association between UGT2B17 mismatch in the GVHD direction and the incidence of acute GVHD, chronic GVHD, relapse, or survival. However, the use of a UGT2B17-positive donor was an independent risk factor for higher transplant-related mortality and lower survival after transplantation. UGT2B17 is a metabolic enzyme for hormones, drugs, and potentially toxic exogenous compounds and is expressed in subsets of haematopoietic cells. Thus, the enzyme function of UGT2B17 in donor cells may affect the outcome of allogeneic HSCT. PMID- 15813851 TI - Increased incidence of EBV-related disease following paediatric stem cell transplantation with reduced-intensity conditioning. AB - The incidence of Epstein-Barr virus (EBV) viraemia and lymphoproliferative disease (LPD) was studied in a consecutive cohort of 128 paediatric patients undergoing stem cell transplantation (SCT) with reduced-intensity conditioning (RIC; n = 65) or conventional-intensity conditioning (CIC; n = 68). Following CIC, six of 68 (8%) developed viraemia; all remained asymptomatic. EBV viraemia (23 of 65 patients = 35%, P < 0.001) and LPD (10 of 65 = 15%, P < 0.001) were significantly more frequent following RIC. Of the 23 RIC patients who developed viraemia, eight remained asymptomatic, five had symptomatic viraemia (fever +/- rash), and 10 patients developed LPD, two of whom died. An absolute lymphocyte count of <0.3 x 10(9)/l at the time of onset of viraemia was strongly predictive of development of LPD (P < 0.05) in this group. The incidence of viraemia was significantly higher in patients receiving serotherapy with antithymocyte globulin (ATG; 15 of 43, 35%) than Campath (12 of 73, 16.4%, P < 0.05). Primary immunodeficiency and acute graft-versus-host disease were associated with EBV viraemia in univariate analysis, but were not independent risk factors. In conclusion, EBV viraemia and LPD appear to be significantly more common in children following RIC SCT, particularly with selective depletion of recipient T cells relative to B cells following the use of ATG. This probably reflects the profound immunosuppression following RIC SCT, together with the incomplete ablation of recipient-derived B cells. PMID- 15813852 TI - Anti-prothrombin IgG from patients with anti-phospholipid antibodies inhibits the inactivation of factor Va by activated protein C. AB - Interference of anti-phospholipid antibodies with the protein C pathway has been suggested to play a role in the development of thrombosis in the anti phospholipid syndrome. We studied the effect of IgG preparations containing anti prothrombin antibodies of 17 lupus anticoagulant-positive patients and 12 controls on the inactivation of factor Va (FVa) by activated protein C (APC) in a system with purified coagulation factors. Test IgG was incubated with human prothrombin, phospholipid vesicles and CaCl(2). Protein S, FVa and APC were added and the APC-dependent loss of FVa activity was monitored over time. The residual amount of FVa remaining after 10 min was 14 +/- 4% (mean +/- SD) when 1.5 mg/ml normal IgG was present and ranged between 17% and 82% with 1.5 mg/ml patient IgG. Twelve patients IgG gave values of residual FVa >22% (i.e. 2 SD above the mean of controls), indicating that APC-mediated inactivation of FVa was significantly inhibited. The inhibition was strictly dependent on the presence of prothrombin, proportional to the concentration of IgG and strongly diminished at a 20-fold higher phospholipid concentration. Most, although not all, IgG containing anti prothrombin antibodies inhibit the APC-catalysed FVa inactivation, which may contribute to the increased risk of thrombosis in patients with the anti phospholipid syndrome. PMID- 15813853 TI - Bone marrow endothelial cells in multiple myeloma secrete CXC-chemokines that mediate interactions with plasma cells. AB - Bone marrow endothelial cells (EC) from patients with multiple myeloma (MM) were found to express and secrete higher amounts of the CXC-chemokines CXCL8/interleukin (IL)-8, CXCL11/interferon-inducible T-cell alpha chemoattractant (I-TAC), CXCL12/stromal cell-derived factor (SDF)-1alpha, and CCL2/monocyte chemotactic protein(MCP)-1 than EC from human umbilical vein (HUVEC), considered as a healthy counterpart. Paired plasma cells and several MM cell lines expressed cognate receptors of each chemokine to a variable extent. When cells were exposed to chemokines, CXCL8/IL-8 and CXCL12/SDF-1alpha stimulated their proliferation and all chemokines stimulated cell chemotaxis. It is suggested that angiogenesis also favours MM progression through the release of CXC-chemokines. PMID- 15813854 TI - Nicotinic acetylcholine receptor-mediated stimulation of endothelial cells results in the arrest of haematopoietic progenitor cells on endothelium. AB - The function of endothelial cells that contribute to the regulation of haematopoietic stem/progenitor cells (HSPC) migration from peripheral blood into bone marrow can be influenced by extrinsic factors including nicotine. Therefore, the effect of nicotine on HSPC extravasation was studied. Using a parallel laminar flow chamber, we demonstrated an increase in the number of HSPC adhering to the nicotine-exposed endothelium under conditions of physiological shear stress in vitro. Nicotine-induced adhesion of HSPC was inhibited by mecamylamine, a non-selective nicotinic acetylcholine receptor (nAchR) antagonist. The enhanced adhesive interactions of HSPC with nicotine-exposed endothelial monolayers coincided with the nicotine-induced activation of endothelial cells. Nicotine induced fast cytoskeletal reorganization and formation of filopodia in endothelial cells through interaction with the non-neuronal nAchR expressed by these cells. In addition, nicotine treatment stimulated rapid phosphorylation of Erk1/2 and p-38 in endothelial cells. Finally, nicotine inhibited the stroma derived factor-1-mediated transendothelial migration of HSPC. Decreased migration of HSPC correlated with diminished matrix metalloproteinase-9 activity secreted by bone marrow cells and decreased expression of CD44 on the surface of endothelial cells. Overall, our data suggest that exposure to nicotine causes endothelial cell dysfunction and leads to the pathological arrest of HSPC on endothelium, interfering with their proper migration process. PMID- 15813855 TI - The immune response to primary EBV infection: a role for natural killer cells. AB - The role of antigen-specific CD3(+)CD8(+) cytotoxic T cells in the control of primary Epstein-Barr Virus (EBV) infection is well established. However, time is required for the antigen-specific immune response to develop and expand. In contrast, innate immune responses, such as natural killer (NK) cells, are considered vital early in the infection process. We analysed the scale, phenotype and function of the NK cell response during symptomatic primary EBV infection, infectious mononucleosis (IM) and showed that NK cell numbers were significantly elevated both at diagnosis of IM and in the first month following diagnosis. There were also significant changes in cell phenotype and function, an increase in the proportion of CD56(bright) cells at diagnosis, and freshly isolated cells showing an enhanced ability to kill EBV-infected cell lines. Moreover, in our cohort of IM patients higher NK cell counts were associated with significantly lower viral load in peripheral blood. Our results suggest NK cells have an important role in the control of primary EBV infection by eliminating infected B cells and augmenting the antigen-specific T cell response via release of immunomodulatory cytokines. The magnitude of the NK cell response may ultimately determine whether primary EBV infection has a clinical outcome. PMID- 15813856 TI - Heterogeneous expression pattern of pro- and anti-apoptotic factors in myeloid progenitor cells of patients with severe congenital neutropenia treated with granulocyte colony-stimulating factor. AB - Apoptosis is accelerated in the myeloid progenitor cells of patients with severe congenital neutropenia (CN). Granulocyte colony-stimulating factor (G-CSF) increases neutrophil numbers in most CN patients. The effect of G-CSF on apoptosis in CN was analysed by apoptosis rate and expression of anti- and pro apoptotic factors. G-CSF-treated patients showed higher apoptosis frequency, lower expression of bcl-2 and bcl-xL, but higher expression of bfl-1/A1 and mcl 1. Caspase 9 was highly expressed in patients and controls after G-CSF administration. Thus, G-CSF acts on apoptosis regulation, but additional mechanisms leading to the increase of neutrophil numbers must be assumed. PMID- 15813857 TI - Teicoplanin-dependent antibodies: detection and characterization. AB - There are only a few reports of thrombocytopenia associated with clinical doses of teicoplanin, a glycopeptide antibiotic used against Gram-positive bacteria. We investigated 39 patients receiving teicoplanin; 31 were thrombocytopenic with platelet counts between 1-105 x 10(9)/l and 8 were not thrombocytopenic. We identified 14 thrombocytopenic cases (45%) and two (25%) non-thrombocytopenic cases with IgG teicoplanin-dependent platelet-reactive antibodies. Use of glycoprotein (GP) capture enzyme-linked immunosorbent assay with platelets and GPIIb/IIIa transfected Chinese Hamster Ovary cells as well as flow cytometry with GP-deficient platelets indicated that the GPIIb/IIIa complex is a major target antigen of these antibodies. PMID- 15813858 TI - Co-inheritance of the Hb Sun Prairie mutation with a point mutation at 5'-UTR in the eastern Indian population. AB - Haemoglobin (Hb) Sun Prairie (alpha2-globin cd130, GCT-->CCT, Ala-->Pro) is detected in three unrelated chromosomes, in association with a C-->T transition in the 5'-untranslated region (UTR), two bases upstream from the translation start site. Reported inversion of alpha/beta-mRNA ratio observed in Hb Sun Prairie mutants might stem from the second mutation and should be investigated. Molecular modelling studies indicate that the 130th residue of alpha-globin faces primarily the central cavity of the molecule and is not in contact with any beta chain residue; further, no significant disruption of the Hb structure because of the Sun Prairie mutation is discernible. Depression of translation because of the second mutation of a conserved base in the 5'-UTR might explain the observed clinical severity. PMID- 15813860 TI - Introduction and comparison of two methods of assessment of coronary lesions in Atlantic salmon, Salmo salar L. AB - This study investigated and compared quantitative and semi-quantitative coronary lesion evaluation in Atlantic salmon, Salmo salar. A total of 121 immature farmed and 47 sexually mature wild Atlantic salmon were included. Coronary arteries from all fish were morphometrically evaluated using a semi-quantitative method. A subsample of 76 salmon was additionally evaluated using a quantitative method. Another subsample of 71 salmon was used for reliability testing of the semi quantitative method. Ten cross-sections of coronary arteries located on the ventral surface of the bulbus arteriosus were semi-quantitatively scored with regard to myointimal coronary lesions. Quantitative measurements comprised maximal intimal thickness and area lesion (%) while semi-quantitative measurements comprised maximal intimal thickness and lesion degree (%). Scores for individual fish were expressed as the average of lesion score, denoted as mean range lesion. One person performed all evaluations blindly. Two pathologists tested repeatability of the semi-quantitative lesion score measurements blindly. Both semi-quantitative and quantitative variables increased significantly with increasing lesion score. Semi-quantitative estimation gave significantly higher maximal intimal thickness values than quantitative measurement. Semi-quantitative measurements were found to explain 63.2% of variation in quantitatively measured maximal intimal thickness. Due to lack of agreement, the two methods are not directly comparable and one method cannot replace the other. Repeatability within and between observers with regard to semi-quantitative classification was very good with Kappa values larger than 61.5%. The semi-quantitative method was a valid and reliable method for coronary lesion evaluation in Atlantic salmon. PMID- 15813861 TI - Isolation and characterization of strains of Flavobacterium columnare from Brazil. AB - Flavobacterium columnare is an important pathogen of freshwater fish, implicated in skin and gill disease, often causing high mortality. An outbreak of skin disease in fingerling and adult Nile tilapia, Oreochromis niloticus (L.), cultivated in a recirculation system, was investigated. Four strains were isolated and characterized by biochemical reactions, enzyme production, fatty acid profile and analysis of the 16S-23S rDNA intergenic spacer region. All strains were identified as F. columnare. Experimental infection assays with one of these strains (BZ-5-02) were conducted and pathogenicity (by intramuscular route) was demonstrated in Nile tilapia and channel catfish, Ictalurus punctatus (Rafinesque). This is the first report of characterization of Brazilian strains of F. columnare. PMID- 15813862 TI - Antigenicity of Streptococcus agalactiae extracellular products and vaccine efficacy. AB - Streptococcus agalactiae is a major bacterial pathogen that is the cause of serious economic losses in many species of freshwater, marine and estuarine fish worldwide. A highly efficacious S. agalactiae vaccine was developed using extracellular products (ECP) and formalin-killed whole cells of S. agalactiae. The vaccine efficacy following storage of S. agalactiae ECP and formalin-killed S. agalactiae cells at 4 degrees C for 1 year was determined. The stored ECP containing S. agalactiae formalin-killed cells failed to prevent morbidity and mortality among the vaccinated fish, and the relative percentage survival was 29. Serum antibody responses of the stored ECP and freshly prepared ECP against soluble whole cell extract of S. agalactiae indicated that significantly less antibody was produced in fish immunized with stored ECP and S. agalactiae cells than in those fish immunized with freshly prepared ECP and S. agalactiae cells at day 31 post-vaccination. Silver staining of sodium dodecyl sulphate polyacrylamide gels and immunostaining of Western blots with tilapia antiserum to S. agalactiae revealed that predominant 54 and 55 kDa bands were present in the freshly prepared ECP fraction. The 55 kDa band was absent from the stored ECP and new bands below 54 kDa appeared on the Western blot. The results of this study on S. agalactiae ECP provide evidence for a correlation between protection and antibody production to ECP and for the importance of the 55 kDa ECP antigen for vaccine efficacy. PMID- 15813863 TI - Cutaneous antibodies from channel catfish, Ictalurus punctatus (Rafinesque), immune to Ichthyophthirius multifiliis (Ich) may induce apoptosis of Ich theronts. AB - This study explored the existence of apoptosis (programmed cell death) in Ichthyophthirius multifiliis Fouquet (Ich) theronts and determined the effect of cutaneous antibodies in skin culture fluid from fish immune to Ich on theront apoptosis. Apoptosis was detected in theronts and was clearly distinguished by fluorescent microscopy after staining with acridine orange and propidium iodide. The apoptotic theronts showed characteristic chromatin condensation and nuclear fragments containing chromatin pieces. The externalization of phosphatidylserine on the plasma membrane of apoptotic theronts was detected with fluorescein isothiocyanate-conjugated annexin using flow cytometry. Theront apoptosis was induced using the skin culture fluid from fish immune to Ich, which contained cutaneous antibodies against Ich. The highest apoptosis appeared in theronts exposed to immune skin culture fluid at a 1:10 dilution, compared with those at 1:20 and 1:40 dilutions. A direct correlation was noted between the percentage of apoptotic theronts and exposure duration to immune skin culture fluid. The study indicated that antibody reaction with theronts (immobilization) played an important role in theront apoptosis, but it could not be excluded that other components released from the excised skin had effects on theronts. PMID- 15813864 TI - Isolation of lymphocystis disease virus from sole, Solea senegalensis Kaup, and blackspot sea bream, Pagellus bogaraveo (Brunnich). AB - Two viruses were isolated from cultured sole, Solea senegalensis, and wild blackspot sea bream, Pagellus bogaraveo, and preliminarily characterized as lymphocystis disease viruses (LCDVs). Viral isolates were characterized by morphological, biochemical and biophysical properties. In addition, the susceptibility of four fish cell lines was also tested. LCDV isolates developed cytopathic effects on the SAF-1 cell line at 5 and 6 days post-infection and reached titres of 10(6) TCID50 mL(-1). The antigenic and structural protein analysis of the two new LCDV isolates showed identical profiles to that obtained for LCDV strain Leetown NFH (ATCC VR-342), used as a reference viral strain, and for an LCDV isolate collected from gilt-head sea bream, Sparus aurata, cultured in southern Spain. Molecular confirmation was performed by polymerase chain reaction. Specific primers for LCDV produced a 270-bp DNA fragment, the expected size for LCDV. PMID- 15813865 TI - Adhesion to sole, Solea senegalensis Kaup, mucus of microorganisms isolated from farmed fish, and their interaction with Photobacterium damselae subsp. piscicida. AB - Abstract Most studies carried out to select microorganisms as candidate probiotics have focused on in vitro antagonism tests, such as the production of inhibitory compounds against pathogenic microorganisms. However, attachment to mucous surfaces could be another criterion to be considered when selecting potential probiotics for aquaculture. Nineteen isolates obtained from farmed Senegalese sole, Solea senegalensis Kaup, and gilthead sea bream, Sparus aurata L., have been evaluated for their capacity to adhere to skin and intestinal mucus of Senegalese sole, and their antagonistic effect against Photobacterium damselae subsp. piscicida, an important pathogen for farmed sole. The isolates from gilthead sea bream showed the highest percentage of adhesion to sole mucus, whilst the pathogenic microorganisms assayed and the isolates from sole showed, in general, a lower ability to adhere to sole mucus. The results suggest that the adhesion to fish mucus was more dependent on the isolate tested than on the host mucus. The isolates from gilthead sea bream also showed a higher antagonistic activity against P. damselae subsp. piscicida than those from Senegalese sole. Four isolates were selected, on the basis of their adhesive ability and antagonistic effect on P. damselae subsp. piscicida, to study their interactions with the pathogen in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions. The results obtained show the ability of three isolates to reduce the adhesion of P. damselae subsp. piscicida to sole mucus under displacement and competition conditions. The adhesion of the pathogen to sole intestinal mucus was also significantly reduced when three isolates were assayed under exclusion conditions. PMID- 15813866 TI - Evidence for disruption of Na(+)-K(+)-ATPase and hsp70 during vibriosis of sea bream, Sparus (=Rhabdosargus) sarba Forsskal. AB - Osmoregulation via the sodium pump (Na(+)-K(+)-ATPase) and cytoprotection via expression of different families of heat shock protein (hsp) were studied at transcriptional/translational levels during progressive vibriosis in silver sea bream, Sparus (=Rhabdosargus) sarba Measurements of Na(+)-K(+)-ATPase activity showed that an early and drastic decline occurred in the kidney of infected fish. This reduction in Na(+)-K(+)-ATPase activity was not caused by transcriptional downregulation of genes coding for either the Na(+)-K(+)-ATPase alpha or beta subunits. Using specific antibodies, data from immunoassays showed that the decreased sodium pump activity was caused by the specific loss of the translated glycosylated Na(+)-K(+)-ATPase beta subunit. Data from immunoassays of different hsp families demonstrated that the expression of hsp90 and hsp60 remained unchanged throughout vibriosis whereas expression of the hsp70 family decreased in kidney and liver tissues. As hsp70 is a multigene family, the expression of the constitutive (hsc70) and inducible (hsp70) members of the hsp70 family were studied and it was found that hsp70 and hsc70 expression decreased from an early stage of infection in the kidney and the liver respectively. Reverse transcriptase-polymerase chain reaction analysis of the hsp70 transcription inducing factor, hsf1, demonstrated that loss of cytoprotective function during vibriosis was mediated by a downregulation of hsf1 transcription. PMID- 15813867 TI - Reporting of adverse events following immunization in Australia. AB - It is an important component of any immunization programme that vaccine safety is monitored by carrying out surveillance for adverse events following immunization (AEFI). Such surveillance can be active or passive. Active surveillance will detect more AEFI, but the vast majority will be minor events. Passive surveillance is probably more appropriate for routine AEFI surveillance, while active surveillance can be used to monitor a new vaccine or to test whether a specific severe event is significantly associated with immunization. Australia has a predominantly passive surveillance system. The system has recently been centralized, providing useful national data on vaccine safety. PMID- 15813868 TI - The epidemiology and pathogenesis of neonatal necrotizing enterocolitis. PMID- 15813869 TI - Epidemiology of necrotizing enterocolitis--Part I: Changing regional trends in extremely preterm infants over 14 years. AB - OBJECTIVES: Advances in perinatal care include exogenous surfactant, unequivocal acceptance of antenatal steroids and in utero and ex utero transfers to tertiary centres. Increased survival of extremely premature infants may change the incidence and outcome of necrotizing enterocolitis (NEC). Our aim was to examine the trends in the incidence of NEC, surgery and mortality in infants of 24-28 weeks gestation in a retrospective regional review of three epochs over a span of 14 years. METHODS: Radiologically or surgically proven NEC cases were determined from the New South Wales Neonatal Intensive Care Unit Study database. Three epochs were examined. A total of 360 infants were admitted in 1986-87 (Epoch 1), 622 in 1992-93 (Epoch 2) and 673 in 1998-99 (Epoch 3). RESULTS: There was an increase in neonatal intensive care unit admissions and a decrease in early and overall mortality of these very premature infants across the epochs. None of the early deaths was due to NEC. The incidence of NEC decreased in post day 5 survivors: 33 cases in Epoch 1 (12%), 60 cases in Epoch 2 (12%) and 34 cases in Epoch 3 (6%, P < 0.001). There was no change in surgical intervention (45%, 57% and 41%, respectively) or mortality due to NEC (37%, 27% and 32%). The reduced incidence of NEC was not singularly influenced by antenatal steroids, exogenous surfactant or outborn delivery. In a multivariate analysis, only later epoch of birth was independently associated with reduced NEC risk. CONCLUSIONS: With improved care and survival of extremely premature infants, the incidence of NEC has decreased, but it remains a disease of high mortality and morbidity. PMID- 15813870 TI - Epidemiology of necrotizing enterocolitis--Part II: Risks and susceptibility of premature infants during the surfactant era: a regional study. AB - OBJECTIVES: We observed a reduced incidence of necrotizing enterocolitis (NEC) despite increased survival of extremely premature infants over the last two decades. A different susceptibility pattern to NEC according to gestation has been proposed. We aim to examine the influence of perinatal risk factors for NEC in infants less than 32 weeks gestation, in our region, in the post-surfactant era. METHODS: NEC incidence, perinatal risk factors and neonatal outcomes were examined in a cohort of 4649 infants of 24-31 weeks gestation from the Neonatal Intensive Care Unit Study (NICUS) database, admitted during 1994-99. RESULTS: A total of 178 (3.8%) infants developed NEC. More 24-27 weeks infants than 28-31 weeks infants had NEC (6.6% vs 2.6%, P < 0.001). Although low gestation infants had higher mortality and surgery need, post-operative survival (60%) was comparable to the higher gestation infants. NEC in the higher gestation group tended to be associated with perinatal risk factors, while risk factors were equally common in low gestation infants who developed NEC or not. Overall, gestation (P < 0.001), small for gestational age (P= 0.008), hyaline membrane disease (P < 0.001), placental abruption (P = 0.002) and earlier year of birth (P = 0.03) were independently associated with increased odds of NEC. Maternal hypertensive disease of pregnancy was associated with reduced NEC risk (P = 0.02). CONCLUSIONS: The incidence of NEC continues to decrease in the surfactant era and the pattern of susceptibility to NEC is based on gestational age grouping and risk factors. The reduced risk associated with hypertensive disease of pregnancy is unexplained. PMID- 15813871 TI - Prone or supine for infants with chronic lung disease at neonatal discharge? AB - OBJECTIVE: To determine whether infants with chronic lung disease (CLD), ready for neonatal unit discharge, maintain cardiorespiratory stability while sleeping supine. METHODS: Subjects were 15 infants born < 32 weeks gestational age (GA) and ready for discharge from the regional tertiary neonatal intensive care unit. Polysomnography recordings of sleep state, heart rate, arterial oxygen saturation, respiratory effort and nasal/oral airflow were taken prone and supine for up to 3 h post feed with the first position randomly allocated. The main outcome measures were oxygen saturation and apnoea hypopnoea index (AHI). RESULTS: Seven infants (median GA 27 weeks, birthweight 945 g) had CLD and eight infants (median GA 29 weeks, birthweight 1160 g) did not. CLD infants were more mature at study than non-CLD infants (median 39 vs 36 weeks, P = 0.019). Neither oxygen saturation nor AHI were position dependent and no group differences were noted with respect to CLD status. There was a significant interaction of GA and sleep position with less-mature infants spending less time in quiet sleep (QS) in supine position (P = 0.006). These less-mature infants also had a higher AHI (P = 0.033). As expected, the AHI and arousal index (AI) were higher in active sleep (P < or = 0.001, P = 0.013, respectively) and mean oxygen saturation was lower (P = 0.001). CONCLUSIONS: The supine position appears appropriate for very preterm infants with CLD going home from the neonatal unit. Respiratory instability on neonatal discharge is more likely to be associated with immaturity than CLD. PMID- 15813872 TI - Antenatal steroids may reduce adverse neurological outcome following chorioamnionitis: neurodevelopmental outcome and chorioamnionitis in premature infants. AB - OBJECTIVE: To examine the effect of antenatal steroid exposure and in utero inflammation on the development of severe intraventricular haemorrhage, periventricular leukomalacia and long-term neurological outcome in infants less than 30 completed weeks gestation. METHOD: Infants less than 30 completed weeks gestation from January 1996 to July 2001 were identified from a prospectively managed database. Placental pathology was reviewed for the presence or absence of chorioamnionitis and funisitis. Infants were divided into three groups depending on the degree of exposure to fetal inflammation (no inflammation, chorioamnionitis only and chorioamnionitis and funisitis). Data relating to gestational age, birthweight, sex, antenatal steroid exposure, surfactant treatment, days of positive pressure ventilation and days of oxygen requirement were collected. Cerebral ultrasound studies were examined for evidence of intraventricular or intraparenchymal echodensity and periventricular leukomalacia. Long-term neurological outcome was assessed by neurological examination for cerebral palsy and by Griffiths Mental Developmental Assessment for general developmental quotient. RESULTS: Two hundred and twenty infants were identified. The mean gestational age was 27.7 weeks and the mean birthweight 1092 g. Seventy-two per cent of mothers had received a complete course of antenatal steroids. The risk of Grade III intraventricular haemorrhage or intraparenchymal echodensity was associated with exposure to in utero inflammation if a complete course of antenatal steroids had not been received (P = 0.002). This association did not exist if a complete course of antenatal steroids was given (P = 0.62). Fourteen infants had cerebral palsy (7%). The presence of cerebral palsy was also associated with in utero inflammation in the absence of complete antenatal steroid cover (P = 0.03) and not in the presence of complete cover (P = 0.59). The mean general developmental quotient on Griffiths Mental Developmental Assessment at 12 months or 3 years was not affected by exposure to in utero inflammation regardless of antenatal steroid exposure. CONCLUSION: Risk of intraventricular haemorrhage or intraparenchymal echodensity and cerebral palsy was associated with in utero inflammation in the absence of a complete course of antenatal steroids. A complete course of antenatal steroids appeared to extinguish any association between in utero inflammation and adverse neurological outcome. PMID- 15813873 TI - Parents' evaluation of developmental status in children born with a birthweight of 1250 g or less. AB - OBJECTIVE: To evaluate the agreement between parental reporting of development of children born very preterm using the Parents' Evaluation of Developmental Status (PEDS) questionnaire and professional assessment by a paediatric developmental team in the detection of sensorineural disability. METHODS: A cross-sectional cohort study of 362 children born in Queensland with a birthweight < or = 1250 g, who were surviving at 2 and 4 years of age corrected for prematurity, was conducted. Parents completed the PEDS questionnaire prior to their child receiving a neurodevelopmental assessment. The level of agreement for sensorineural disability between the neurodevelopmental assessment and the parents' score on the PEDS questionnaire was measured using the kappa statistic, and screening test characteristics were calculated. Logistic regression was used to investigate factors that might affect agreement. RESULTS: Two hundred and eighty-three (78%) of the eligible children were located and contacted. Of these, 216 (76%) agreed to participate in the study (110, 2-year-olds; 106, 4-year olds). Agreement between the two forms of rating sensorineural disability (developmental quotient > -2SD (standard deviation), cerebral palsy, bilateral blindness and deafness requiring aids) for the 4-year age group children was fair (kappa = 0.27, P = 0.001). PEDS accurately identified 69% (11 of 16) of disabled children and 72% (65 of 90) non-disabled children. The test characteristics for these children were similar to Glascoe's norming sample with a PPV 31% (95% CI: 14-48%), specificity 72% (95% CI: 62-81%), but lower sensitivity 69% (95% CI: 62 81%) and higher false-negative rate 31% (95% CI: 11, 59). Agreement for the 2 year age group was fair with poor test characteristics. Other comparisons for both age groups (PEDS A and B compared using a disability status with DQ > -1SD) showed poor agreement and test characteristics. Gestation age < 27 weeks and maternal education at or below grade 10 in the 2-year age group were the only factors independently affecting agreement. CONCLUSIONS: The agreement between parental evaluation of sensorineural disability status using PEDS and paediatrics developmental assessment in children born very preterm at 4-years corrected age for prematurity compares favourably with Glascoe's norming sample. The lower agreement seen in the 2-year age group limits the utility of PEDS to be used as a screen for disability at an age when early intervention may be useful. The PEDS questionnaire is designed and normed for the general paediatric population, and it is not clear if parents of children born very preterm may have interpreted the PEDS questionnaire in relation to their satisfaction with their child's developmental progress rather than their child's functional ability. PMID- 15813874 TI - The distraction test: the last word? AB - OBJECTIVE: Universal Newborn Hearing Screening (UNHS) programmes have been widely implemented, but their costs, benefits and long-term logistics remain to be clearly defined. There are few rigorous evaluations of alternative strategies. In this paper, we evaluate the performance of the distraction test component of the two-tiered Victorian Infant Hearing Screening Program (VIHSP). METHODS: All babies born in the State of Victoria, Australia in 1993 who survived the neonatal period were screened for the presence of risk factors for hearing loss. Those at risk were referred for Auditory Brainstem Evoked Response (ABR) screening by a professional audiologist. All others were screened by modified distraction test at age 7-9 months. This birth cohort was followed through age 6 for diagnoses of congenital hearing loss resulting in fitting of hearing aids. Estimates of false positives, false-negatives, sensitivity, specificity and positive predictive values were determined for the distraction test as a population screen. Ages at diagnosis and aid fitting for screen failures with hearing loss were compared with current goals. RESULTS: For targeted (moderate or greater-aided) losses, the distraction test yielded eight (0.02%) documented false-negatives (one severe and seven moderate) and an estimated 4265 (99%) false-positives. Distraction test sensitivity was 65%, specificity 91% and PPV 0.3%. Mean age at diagnosis for distraction test failures across all severities, including mild losses, was 23 (SD 18) months with a mean age at aid fitting of 26 (SD 20) months. CONCLUSIONS: The distraction test screen generated large numbers of false-positives and a significant number of false-negatives, performing particularly poorly with moderate losses. Ages at diagnosis and aid fitting for screen failures were far older than currently accepted goals. There is little evidence that the distraction test can be made to work acceptably as a population-based screen. PMID- 15813875 TI - Current issues regarding the use of pneumococcal conjugate and polysaccharide vaccines in Australian children. AB - OBJECTIVE: To present the results of child pneumococcal vaccination studies in the setting of current Australian disease epidemiology and immunization policy, and issues that clinicians should consider in discussions with families. METHODS: This paper includes a narrative review of randomized, controlled, double blind studies and systematic reviews which evaluated the efficacy of child pneumococcal vaccination. RESULTS: 7PCV is expected to prevent > 80% of childhood invasive pneumococcal disease (IPD, includes meningitis, septicaemia/bacteraemia) and the associated mortality. 7PCV may prevent 6% of all pneumonia, 18% of radiographically-defined pneumonia, 6% of all otitis media (OM) and 20%-40% of tympanostomy tube procedures. It may also reduce IPD due to antibiotic-resistant pneumococci, and prevent IPD in unvaccinated individuals. The impact of replacement disease caused by non-vaccine serotypes is not yet known. Pneumococcal polysaccharide vaccines given to 2-year-old children may prevent approximately 19% of all and 26% of recurrent OM. CONCLUSION: The Australian Standard Vaccination Schedule recommends universal infant immunization with seven valent pneumococcal conjugate vaccine (7PCV). Universal infant 7PCV will prevent pneumococcal diseases and deaths. The potential for its impact to be reduced in the long-term by serotype replacement must be closely monitored. Information concerning disease epidemiology, vaccine efficacy and safety, disease risk perception and national costs may prove useful in discussions with families. PMID- 15813876 TI - Physician counselling practices and decision-making for extremely preterm infants in the Pacific Rim. AB - OBJECTIVES: This study was undertaken to evaluate physician counselling practices and resuscitation decisions for extremely preterm infants in countries of the Pacific Rim. We sought to determine the degree to which physician beliefs, parents' opinion and medical resources influence decision-making for infants at the margin of viability. METHODS: A survey was administered to neonatologists and paediatricians who attend deliveries of preterm infants in Australia, Hong Kong, Japan, Malaysia, Taiwan and Singapore. Questions were asked regarding physician counselling practices, decision-making for extremely preterm infants and demographic information. RESULTS: Physicians counsel parents antenatally with increasing frequency as gestational age increases. Most physicians discuss infant mortality and morbidity with parents prior to delivery. Physicians less frequently discuss the option of no resuscitation of an extremely preterm infant, withdrawal of support at a later time, or financial costs to parents. Severe congenital malformations, perception of a poor future quality of life, parental wishes and a high probability of death for the infant are influential in limiting resuscitation in very preterm infants for a majority of physicians. Less influential factors are parent socioeconomic status, language barriers, financial costs for the family, allocation of national resources, moral or religious considerations, or fear of litigation. Physician thresholds for resuscitation of infants ranged between 22 and 25 weeks gestation and between 400 and 700 g birthweight. CONCLUSIONS: We report physician beliefs and practices regarding resuscitation and the counselling of parents of extremely preterm infants in Pacific Rim countries. While we find variation among countries, physician practices appear to be determined by ethical decision-making and medical factors rather than social or economic factors in each country. PMID- 15813877 TI - Delayed infant death following catastrophic deterioration during breast-feeding. PMID- 15813878 TI - Hepatic failure in a child with anti-epileptic hypersensitivity syndrome. AB - An 11-year-old boy developed severe hypersensitivity reaction to phenobarbitone resulted in fulminant hepatic failure. During the course of illness, he developed clinical features compatible with severe acute respiratory syndrome (SARS) that may have complicated the recovery of his underlying hypersensitivity reaction, which was subsequently controlled with intravenous immune globulin and corticosteroids. PMID- 15813879 TI - Cerebral thrombosis in childhood nephrosis. AB - Sinovenous thrombosis is an uncommon but serious complication associated with nephrotic syndrome in children. We describe a 9-year-old Caucasian boy who presented with dehydration, vague neurological symptoms and seizures. A diagnosis of nephrotic syndrome was made during the course of hospitalization. The serum antithrombin III level was decreased and brain imaging showed cerebral sinovenous thrombosis. Anticoagulant therapy with heparin was commenced and the patient made a slow but gradual clinical, as well as radiological recovery. We describe this case and review available literature to highlight the importance of suspecting and recognizing this potentially life threatening complication and initiating early treatment. PMID- 15813880 TI - Autoimmune hepatitis/primary sclerosing cholangitis overlap syndrome in a child: diagnostic usefulness of magnetic resonance cholangiopancreatography. AB - A 5-year-old Chinese girl with 1-year history of progressive jaundice, steatorrhoea and pruritus was referred. Physical examination showed failure to thrive, marked jaundice, finger clubbing and hepatomegaly. There was laboratory evidence of cholestatic jaundice and autoimmunity, with marked elevation of alkaline phosphatase (ALP) and gamma-glutamyl transferase (gammaGT). Histology of percutaneous liver biopsy revealed hepatitis around the portal triad, as well as features of liver cirrhosis. Primary sclerosing cholangitis (PSC) overlapping with autoimmune hepatitis (AIH) was suspected. Endoscopic retrograde cholangiopancreatography (ERCP) was not feasible as there was no weight appropriate ERCP scope available. Magnetic resonance cholangiopancreatography (MRCP) was performed and revealed areas of irregularity and slight attenuation of the right and left hepatic ducts, representing stricturing, in keeping with PSC. PSC/AIH overlap syndrome was diagnosed in this child in which MRCP has contributed to its diagnosis. PMID- 15813881 TI - Cardiac pacing in the management of severe pallid breath-holding attacks. AB - Reflex anoxic seizures (pallid breath-holding attacks) can be managed with reassurance in the majority of individuals. In a minority of cases where frequent syncopal and seizure activity occurs, intervention needs to be considered. We report a case of a 19-month infant with a history of severe reflex anoxic seizures who underwent pacemaker insertion with a spectacular result, with complete termination of syncope and seizures, and improvement in quality of life for the patient and family. A literature review of the safety and effectiveness of pacemaker insertion is also presented to support its use as a treatment option for this condition. PMID- 15813883 TI - Intratemporal complications of acute otitis media in children. PMID- 15813884 TI - Rural paediatrics and training in 'new morbidity'. PMID- 15813889 TI - One pot conjugation of the polypeptides directed by phosphorus oxychloride. AB - A series of homopeptides and their conjugates were synthesized in one pot reaction in the presence of phosphorus oxychloride and the conjugate yield was structurally dependent. Menthol and benzylamine conjugated to the homopeptides quantitatively. Homopeptides when treated with diisopropyloxyphosphite (DIPPH) and NaClO yield the corresponding N-phosphoryl peptides. Electrospray ionization mass spectrometry (ESI-MS)/MS was used to study the structure of peptide conjugates. This paper reports a simple method to synthesize the homopeptides and their conjugate derivatives and the fact that phosphoryl peptides could also be obtained by one pot reaction. PMID- 15813890 TI - Comparison of assays for determination of peptide content for lyophilized thymalfasin. AB - Precise determination of the peptide content in drug substance samples depends highly upon the particular peptide compound and methodology used. Four independent methods were evaluated and compared to determine which would produce the best experimental precision for analysis of thymalfasin (thymosin alpha-1). Four different methods were evaluated including elemental analysis (CHN), quantitative amino acid analysis (AAA), high-performance liquid chromatography (HPLC), and Kjeldahl. This study demonstrates that the AAA method is highly variable in one laboratory while quite precise in another laboratory. Similarly, HPLC results depended on the laboratory conducting the study with more precise values obtained under cGMP. On the contrary, the CHN method yielded highly precise [i.e. <2% coefficient of variation (CV)] values. As precise knowledge of protein content is fundamental for the compounding of final pharmaceutical product of a specific potency, the CHN analysis is recommended for peptide content determination of the drug substance thymalfasin. PMID- 15813891 TI - Synthesis and evaluation of a new series of peptide-based endothelin receptor antagonists. AB - Novel peptide-based endothelin (ET) receptor antagonists were designed and synthesized in our laboratory. BQ-485, HIM-CO-Leu-d-Trp-d-Trp-OH, was selected as the leading compound. The primary structures of these new tripeptides were ABO-CO Leu-d-Trp-d-AA(X)-OH. The introduction of unnatural aromatic amino acids into these tripeptides was useful in the structure-activity relationship studies. Among the 20 tripeptides, 16 of them showed high activities against the contraction of rat aortic smooth muscles induced by ET-1. PMID- 15813892 TI - Efficacy of the phosphorylation of synthetic peptides by purified catalytic subunit of PKA (PKAcat) from bovine lens depends on the amino acid sequence of the peptides. AB - Protein kinase (PK) A catalytic (PKAcat) subunit was purified to homogeneity from bovine lens using a 100-kDa cut-off membrane filtration followed by different chromatographic procedures. The molecular weight of PKAcat was found to be 41 kDa. The kinase phosphorylates histone IIIs and other synthetic modified peptides of VRKRTLRRL with different amino acid environment. The extent of phosphorylation depends not only on the presence of Ser or Thr (phosphorylating residues) but also on other surrounding amino acid residues. Although some peptides compete in phosphorylating histone, they are not very significant. The result suggests that the extent of phosphorylation depends on the amino acid residue(s) surrounding phosphorylable residue(s) on the peptide. PMID- 15813893 TI - Biosynthesis and purification of a hydrophobic peptide from transmembrane domains of G-protein-coupled CB2 receptor. AB - A major challenge for the structural study of the seven-transmembrane G-protein coupled receptors is to obtain a sufficient amount of purified protein at the milligram level, which is required for either nuclear magnetic resonance (NMR) spectroscopy or X-ray crystallography. In order to develop a high-yield and cost effective method, and also to obtain preliminary structural information for the computer modeling of the three-dimensional receptor structural model, a highly hydrophobic peptide from human cannabinoid subtype 2 receptor CB2(65-101), was chosen to develop high-yield membrane protein expression and purification methods. The peptide included the second transmembrane helix with the associated loop regions of the CB2 receptor. It was over-expressed in Escherichia coli, with a modified TrpDelta LE1413 (TrpLE) leading fusion sequence and a nine-histidine tag, and was then separated and purified from the tag in a preparative scale. An experimental protocol for the chemical cleavage of membrane protein fragment was developed using cyanogen bromide to remove the TrpLE tag from the hydrophobic fusion protein. In addition, protein uniformly labeled with isotopic 15N was obtained by expression in 15N-enriched minimum media. The developed and optimized preparation scheme of expression, cleavage, and purification provided a sufficient amount of peptide for NMR structure analysis and other biophysical studies that will be reported elsewhere. The process of fusion protein cleavage following purification was monitored by high-performance liquid chromatography (HPLC) and mass spectrometry (MS), and the final sample was validated by MS and circular dichroism experiments. PMID- 15813894 TI - Binding of endomorphin-2 to mu-opioid receptors in experimental mouse mammary adenocarcinoma. AB - Endomorphin-2 (Tyr-Pro-Phe-Phe-NH2) binds with high affinity and selectivity to the mu-opioid receptor. In the present study, [125I]endomorphin-2 has been used to characterize mu-opioid-binding sites on transplantable mouse mammary adenocarcinoma cells. Cold saturation experiments performed with [125I]endomorphin-2 (1 nM) show biphasic binding curves in Scatchard coordinates. One component represents high affinity and low capacity (K(d) = 18.79 +/- 1.13 nM, B(max) = 635 +/- 24 fmol/mg protein) and the other shows low affinity and higher capacity (K(d) = 7.67 +/- 0.81 microM, B(max) = 157 +/- 13 pmol/mg protein) binding sites. The rank order of agonists competing for the [125I]endomorphin-2 binding site was [d-1-Nal3]morphiceptin > endomorphin-2 >> [d Phe3]morphiceptin > morphiceptin > [d-1-Nal3]endomorphin-2, indicating binding of these peptides to mu-opioid receptors. The uptake of 131I-labeled peptides administered intraperitoneally to tumor-bearing mice was also investigated. The highest accumulation in the tumor was observed for [d-1-Nal3)morphiceptin, which reached the value of 8.19 +/- 1.14% dose/g tissue. PMID- 15813895 TI - Analogues of arginine vasopressin and its agonist and antagonist modified in the N-terminal part of the molecule with l-beta-homophenylalanine. AB - In continuation of our efforts to elucidate the role of positions 2 and 3 in arginine vasopressin (AVP) and its analogues, we designed and synthesized peptides modified in these positions with l-beta-homophenylalanine (beta-Hph). Two of them had just this single modification, the next two peptides are analogues of the V2 agonist, namely [3-mercaptopropionic acid (Mpa)1]AVP (dAVP). The last two compounds were designed by substitution of positions 2 or 3 of a potent V(1a) antagonist, [1-mercaptocyclohexaneacetic acid (Cpa)1]AVP, with beta Hph. All the peptides were tested for their pressor and antidiuretic and uterotonic in vitro activities in the rat. All the activities tested have been found to be significantly decreased. Three analogues, i.e. [Mpa(1),beta-Hph2]AVP, [Cpa1,beta-Hph2]AVP, [Cpa1,beta-Hph3]AVP, turned out to be uterotonic antagonists with pA2 = 6.3 +/- 0.2, 6.3 +/- 0.1, 6.0 +/- 0.3 respectively. The last one exhibited antipressor properties also (pA2 = 6.4 +/- 0.1). PMID- 15813897 TI - Melatonin-estrogen interactions in breast cancer. AB - In this article, we review the experimental data supporting an oncostatic role of melatonin on hormone-dependent mammary tumors. Beginning with the evidence on the role of estrogens in breast cancer etiology and mammary tumor growth, we summarize the actual therapeutic strategies with estrogens as a target. Additionally, we demonstrate that melatonin fulfills all the requirements to be considered as an antiestrogenic drug which shares properties with drugs of the two main pharmacological groups of substances which interact with the estrogen signaling pathways such as: (i) drugs that act through the estrogen receptor interfering with the effects of endogenous estrogens; and (ii) drugs that interfere with the synthesis of estrogens by inhibiting the enzymes controlling the interconversion from their androgenic precursors. Furthermore, melatonin decreases circulating levels of estradiol. These three antiestrogenic mechanisms suggest that melatonin may have an important role in the prevention and treatment of hormone-dependent mammary cancer. PMID- 15813898 TI - Exogenous melatonin enhances bile flow and ATP levels after cold storage and reperfusion in rat liver: implications for liver transplantation. AB - Although the use of melatonin in the transplantation field has been suggested, it has not been previously tested in a liver cold-storage model. We used a rat liver model to study (a) the dose-dependent effect of melatonin on bile production, and (b) the potential of melatonin to improve liver function after cold-storage. Male Wistar rats were perfused with Krebs-Henseleit bicarbonate buffer (KHB) at 37 degrees C without or with 25, 50, 100 and 200 microM melatonin. Each dose of melatonin stimulated bile production. For cold-storage studies, livers were flushed with either University of Wisconsin (UW) or Celsior solution and stored for 20 hr at 4 degrees C. Reperfusion (120 min) was performed with KHB at 37 degrees C. In subsequent studies, 100 microM melatonin were added to the perfusate during the reperfusion period. ATP and melatonin levels in the tissue were measured. Bile analysis was performed by measuring melatonin, bilirubin and gamma-glutamyl transpeptidase (gamma-GT) levels in the fluid. A dose-dependent increase in bile secretion, associated with an enhanced melatonin and bilirubin levels in the bile were observed. Also, tissue levels of melatonin increased in a dose-dependent manner. When melatonin was added during the reperfusion period, bile production and bile bilirubin levels increased both with UW and Celsior solutions. The analysis of gamma-GT in the bile showed an increase in the Celsior preserved liver and the addition of melatonin to the perfusate reduced this effect. Tissue ATP levels were higher when melatonin was added to the perfusion medium. Higher levels of melatonin in bile than in tissue were found. In conclusion, we demonstrate that melatonin improves significantly the restoration of liver function after cold-storage and reperfusion. PMID- 15813899 TI - Differential regulation of estrogen receptor alpha, glucocorticoid receptor and retinoic acid receptor alpha transcriptional activity by melatonin is mediated via different G proteins. AB - Melatonin has been shown to bind to the MT1 G protein-coupled receptor (GPCR) in MCF-7 breast cancer cells to modulate the estrogen response pathway suppressing estrogen-induced estrogen receptor alpha (ERalpha) transcriptional activity, blunting ER/DNA binding activity and suppressing cell proliferation. In these studies we have examined the effect of melatonin on the transcriptional activity of the ERalpha and other members of the steroid/thyroid hormone receptor superfamily, namely, the glucocorticoid receptor (GR) and the retinoic acid receptor alpha (RARalpha). As with the ERalpha, melatonin represses ligand (dexamethasone)-induced activation of the GR. This effect of melatonin on ERalpha and GR is blocked by pertussis toxin (PTX) suggesting that melatonin's actions may be mediated via a PTX-sensitive G(alphai) protein. In contrast, melatonin potentiates the action of all-trans-retinoic acid on RARalpha transcriptional activation and enhances RARalpha/DNA binding activity, an action which is not PTX sensitive. Expression of a dominant-positive G(alphai2) protein, with which the MT1 receptor has been shown to couple, is able to mimic the effect of melatonin on ERalpha but not RARalpha transcriptional activation in breast cancer cells. This demonstrates that GPCRs can modulate the transcriptional activity of various steroid receptors in response to their ligand through activation of different G protein signaling pathways. PMID- 15813900 TI - Effect of melatonin administration on parameters related to oxidative damage in hepatocytes isolated from old Wistar rats. AB - Aging induces changes in several organs and tissues, such as the liver, and this process might be due to oxidative damage caused by free radicals and inflammatory mediators. Melatonin is a secretory product with well-known antioxidant properties. The aim of this study was to investigate the effect of melatonin administration on age-induced alterations in hepatocytes. Twenty-two-month old male Wistar rats were treated with oral melatonin for 10 wk. At the end of the treatment, hepatocytes were isolated and cultured, and different parameters were measured in both cells and medium. Aging induced a significant increase in lipid peroxidation, nitric oxide, carbon monoxide and cyclic guanosyl-monophosphate, as well as a reduction in adenosine triphosphate content and phosphatidylcholine synthesis when compared to young animals. Melatonin administration significantly ameliorated all these age-related changes in males. Melatonin administration seems to exert beneficial effects against age-induced changes in hepatocytes. PMID- 15813901 TI - First insights into regiospecific transnitrosation reactions between tryptophan derivatives: melatonin as an effective target. AB - Melatonin, a derivative of the essential amino acid tryptophan, has been portrayed as a hormone, a tissue factor, an autocoid, a paracoid, and a vitamin with antioxidative capabilities. In the present study a novel reaction which cannot be attributed to any of these suggested features, i.e. the transfer of the nitroso-function from N-nitrosotryptophan derivatives to melatonin, is unequivocally demonstrated. In the lipophilic buffer dimethylsulfoxide reaction of N-acetyl-N-nitrosotryptophan (NANT) with melatonin was very slow (k = 1.5 x 10(-6)/m/s), but reversible as shown by 15N-NMR spectrometry. These measurements demonstrated also that the thermodynamical equilibrium lies on the side of N nitrosomelatonin (NOMela). Quantum-chemical calculations performed with the third generation density functional B97-2 additionally predicted that this is also the case in an aqueous environment. In fact, reaction of melatonin with either NANT or N-nitrosotryptophan located at the endothelin-1 fragment 16-21 yielded NOMela with a rate constant of 1.7 +/- 0.5/m/s as shown by capillary zone electrophoresis. Interestingly, the known reactive nitrogen oxide species scavenger, piperazine, did not inhibit the NANT-dependent nitrosation of melatonin, thus very strongly indicating a direct transnitrosation reaction. All of these capabilities are known from the reaction of S-nitrosothiols with thiolate anions and are believed to be highly important in the transport and targeting of nitric oxide. PMID- 15813902 TI - A comparison of the action of amifostine and melatonin on DNA-damaging effects and apoptosis induced by idarubicin in normal and cancer cells. AB - Amifostine is a well-known cell protector and its actions involve free radical scavenging, which is also considered as a mechanism underlying the protective actions of melatonin, a secretory product of the pineal gland. In this work we compared the action of 14 mM amifostine and 50 microM melatonin on DNA damage and apoptosis induced by idarubicin in normal human lymphocytes, leukemic K562 cells and HeLa cancer cells. We employed the alkaline comet assay and pulse-field gel electrophoresis to estimate DNA damage. Apoptosis was evaluated by caspase 3 activity assay assisted by the comet assay to evaluate DNA fragmentation and DAPI staining for detection of morphological changes in chromatin. We found that idarubicin induced apoptosis in normal and cancer cells and its level was correlated with the extent of DNA strand breaks. Amifostine reduced apoptosis and DNA damage in normal cells, but it potentiated these effects in cancer cells in this in vitro study. Melatonin protected both normal and cancer cells against genotoxic treatment and apoptosis induced by idarubicin. We conclude that despite its recognized potential as an antioxidant, melatonin should be considered with caution when used in combination with cancer chemotherapy agents, especially in the case of leukemias. PMID- 15813903 TI - Tryptophan affects both gastrointestinal melatonin production and interrenal activity in stressed and nonstressed rainbow trout. AB - The present experiments were designed to test the hypothesis that elevated dietary levels of l-tryptophan (Trp) result in elevated plasma levels of melatonin and that this increase in plasma melatonin concentration is caused by elevated melatonin production and secretion by the gastro-intestinal-tract (GIT). Feeding juvenile rainbow trout (Oncorhynchus mykiss) Trp-supplemented feed for 7 days resulted in elevated daytime plasma levels of melatonin and reduced poststress plasma cortisol concentrations. Nighttime plasma melatonin concentrations were, however, not affected by elevated dietary Trp. Moreover, stress caused a reduction in daytime plasma levels of melatonin in fish fed Trp supplemented feed, an effect that was counteracted by treatment with an alpha receptor antagonist. These results clearly suggest that elevated dietary intake of Trp results in an increase in the GIT production of melatonin in rainbow trout. A suggestion that was further supported by the results from an in vitro experiment demonstrating that addition of Trp to the incubation medium stimulates melatonin production and release by incubated rainbow trout GIT. The results from this study led us to suggest a possible mechanism for melatonin in mediating the effects of elevated dietary Trp on poststress plasma cortisol concentrations and aggressive behavior in rainbow trout. PMID- 15813904 TI - Melatonin ameliorates bladder damage induced by cyclophosphamide in rats. AB - Cyclophosphamide (CP), an alkylating antineoplastic agent, has potential urotoxicity including causing hemorrhagic cystitis (HC). HC is now accepted as a non-infectious inflammation and the pathogenesis of HC includes cytokine production which leads to inducible nitric oxide synthase (iNOS) induction. Moreover, overproduction of reactive oxygen species (ROS) during inflammation leads to extensive oxidative stress, cellular injury and apoptosis/necrosis via several mechanisms. Based on these facts, the aim of this study was to evaluate the protective effects of melatonin as an antioxidant, iNOS inhibitor and peroxynitrite scavenger against CP-induced urinary bladder damage. A total of 30 male Sprague-Dawley rats were divided into four groups. Three groups received a single dose of CP (100 mg/kg) intraperitoneally with the same times. Group 2 received CP only, group 3 received 5 mg/kg/day and group 4 received 10 mg/kg/day melatonin before and the day after CP administration. Group 1 served as the control. Increased iNOS induction, bladder malonyldialdehyde (MDA) levels and urinary nitrite-nitrate excretion were encountered in the CP-only group leading to severe cystitis. Melatonin exhibited significant protection against CP-induced cystitis by diminishing bladder oxidative stress and blocking iNOS and peroxynitrite production. Oxidants may have a major role in the pathogenesis of CP-induced cystitis and iNOS is an important mediator leading to peroxynitrite production. Melatonin ameliorates bladder damage induced by CP. PMID- 15813905 TI - Pinealectomy impairs adipose tissue adaptability to exercise in rats. AB - This study investigated the effects of pinealectomy and exercise training on rat adipose tissue metabolism. Pinealectomized (PINX) and sham-operated (CONTROL) adult male Wistar rats were subdivided into four subgroups, including PINX untrained, PINX trained, CONTROL untrained and CONTROL trained. At the end of the training period (8 wk), the rats were killed and peri-epididymal adipocytes were isolated for in vitro insulin-stimulated glucose uptake, conversion of D-[U-14C] glucose, l-[U-14C]-lactate, [2-14C]-acetate and [1-14C]-palmitate into 14CO2, and insulin binding. Pinealectomy resulted in a significantly decreased insulin stimulated glucose uptake in adipocytes without affecting insulin-binding capacity. However, in intact control animals only, training promoted a higher baseline glucose uptake in adipocytes. Training influenced the adipocyte ability to oxidize the different substrates: the rates of glucose and palmitate oxidation increased while the rates of lactate and acetate diminished. Nevertheless, these effects of exercise training were not seen in pinealectomized rats. Additionally, an increase in palmitate oxidation was observed in sedentary pinealectomized animals. In conclusion, these data show that the pineal gland alters the patterns of substrate utilization by the adipocyte, in such a way that its absence disrupts the ability to adapt to the metabolic demands evoked by exercise training in rats. PMID- 15813906 TI - Pinealectomy exaggerates and melatonin treatment suppresses neuroma formation of transected sciatic nerve in rats: gross morphological, histological and stereological analysis. AB - At present, an intensive effort for prevention of neuroma formation following peripheral nerve section continues. It has been recently suggested that surgical pinealectomy (Px) induces elevation of the collagen content in the granulation tissue of a wound, while melatonin application after Px suppresses elevation of the collagen accumulation in the tissue. The aim of the present study was to assess whether melatonin had the ability to suppress collagen production and neuroma formation following peripheral nerve transection. A total of 40 male rats (four groups of 10) were left intact (intact controls) or sham operated (sham group), were Px, or were Px and given melatonin (Px + melatonin group). All animals underwent a surgical intervention consisting of right sciatic nerve neurectomy. After 4 wk, the animals were killed following intracardiac perfusion. Gross morphology of neuroma formation in the proximal nerve segment was examined and proximal neuroma evaluated. Macroscopic and microscopic findings revealed that Px caused a proliferation of connective tissue and large neuroma formation at the proximal ends of transected nerves. Stereological analysis showed that there was a statistically significant reduction in connective tissue content of the same region in Px animals treated with melatonin (P < 0.005). The results achieved in a rodent model of sciatic nerve neuroma formation showed that there was a positive correlation between macroscopic and microscopic observations, and that melatonin enhanced axonal regeneration presumably due to its inhibitory effect on neuroma formation. PMID- 15813907 TI - Localization and interactions of melatonin in dry cholesterol/lecithin mixed reversed micelles used as cell membrane models. AB - The state of melatonin confined in dry cholesterol/lecithin mixed reversed micelles dispersed in CCl4 was investigated using 1H-NMR and FT-IR spectroscopies as a function of the melatonin to lecithin molar ratio (R(MLT)) and of the cholesterol to lecithin molar ratio (R(CHL)). An analysis of experimental results leads to the hypothesis that, independent of R(MLT) and as a consequence of anisotropic melatonin/lecithin, melatonin/cholesterol and cholesterol/lecithin interactions, melatonin is totally solubilized in reversed micelles. Melatonin is mainly located in and oriented in the nanodomain constituted by the hydrophilic groups of cholesterol and lecithin. A competition of melatonin and cholesterol for the hydrophilic binding sites of the reversed micelles was observed by changing the R(CHL). Some possible biological implications of the specific interactions governing the solubilization process, the preferential location and the peculiar properties of melatonin confined in cholesterol/lecithin mixed reversed micelles are discussed. PMID- 15813909 TI - A novel mechanism in suppression of erythropoiesis during inflammation: a crucial role of RCAS1. AB - A novel human tumor-associated antigen, receptor-binding cancer antigen expressed on SiSo cells (RCAS1), induces apoptosis in normal human erythroid progenitor cells, which express putative RCAS1 receptors. In the present study, we investigated a possible role of RCAS1 produced by human peripheral blood monocytes (CD14-positive cells) and monocyte-derived macrophages. RCAS1 was immunohistochemically detected in monocytes as well as macrophages. When macrophages were stimulated with lipopolysaccharide (LPS), the expression of RCAS1 was remarkably enhanced. An increased production of RCAS1 mRNA was observed in LPS-stimulated macrophages by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Soluble RCAS1 molecules were only detected in the culture supernatants obtained from LPS-stimulated macrophages. Moreover, LPS stimulated macrophages induced cell death of erythroid progenitor cells through RCAS1 production. These results suggest that macrophages may negatively regulate erythropoiesis at least in part through the production of RCAS1 molecules, and this may contribute to the pathogenesis of the anemia seen in patients with inflammatory disorders. PMID- 15813910 TI - Protein deficiency balance as a predictor of clinical outcome in hereditary spherocytosis. AB - Vertical and horizontal interactions between membrane constituents account for integrity, strength and deformability of the erythrocyte. Disruption of vertical interactions caused by membrane protein deficiencies in hereditary spherocytosis (HS), favor membrane vesiculation with development of spherocytic cells. Our aim was to evaluate the hematological and clinical presentation of HS according to the type and amount of protein deficiency. We studied 81 Portuguese individuals, 71 belonging to 21 families plus 10 unrelated subjects, and found that 51 of them were HS patients. Patients were classified as presenting mild, typical or severe HS, according to laboratory results and clinical follow-up. We performed screening tests and the standardized electrophoretic membrane protein analysis to identify and quantify protein deficiencies. We found band 3 and ankyrin deficiencies as the major causes for HS. The ratios between the value of the primary and/or secondary protein deficiencies showed significantly different values according to the severity of HS, and a significant inverse correlation with the severity of HS was observed. In mild HS, the ratios between protein deficiencies reflected equivalent protein deficiencies, while an unbalance was observed in typical HS, which was enhanced in severe HS. Our data suggest that the relative quantification of each major membrane protein and of the ratios between the values of protein deficiencies may be helpful in providing additional data about the clinical outcome of HS. PMID- 15813911 TI - Haemoglobin and anaemia in a gender perspective: the Tromso Study. AB - OBJECTIVES: To examine the gender-specific distribution of haemoglobin (Hb) and the World Health Organization (WHO) criteria for anaemia compared with the 2.5 percentile for Hb. METHODS: A population-based study from Tromso, Northern Norway. All inhabitants above 24 yr were invited. In total, 26 530 (75%) had their Hb analysed. RESULTS: The 2.5-97.5 percentile of Hb was 129-166 and 114-152 g/L for all men and women, respectively. In men, mean Hb decreased from 148 to 137 g/L between 55-64 and 85+ yr. In women, mean Hb increased from 132 to 137 g/L between 35-44 and 65-74 yr and then decreased to 131 g/L among the oldest. Using the WHO criteria for anaemia (Hb: <130 and <120 g/L, men and women respectively), the prevalence of anaemia in men increased with age from 0.6% aged 25-34 to 29.6% aged 85+. For women, the prevalence of anaemia varied from 9.1%, 2.2% and 16.5% in the age groups of 35-44, 55-64 and 85+ yr, respectively. The WHO criteria gave a two to three times higher prevalence of anaemia compared with the 2.5 percentile of Hb in women, but the difference was small in men. Poor self-rated health was not associated with low values of Hb in women. In men, there was an association in some age groups. CONCLUSION: The WHO criteria for anaemia and the 2.5 percentile for Hb corresponded well for men, but not for women. The WHO criteria of anaemia may result in medicalization of healthy women. PMID- 15813912 TI - Prenatal diagnosis of recessive congenital methaemoglobinaemia type II: novel mutation in the NADH-cytochrome b5 reductase gene leading to stop codon read through. AB - A case of type II recessive congenital methaemoglobinaemia (RCM) observed in a Lebanese subject with a novel mutation in NADH-cytochrome b5 reductase gene is described. A homozygous mutation CAC to AA identified at Thr 295 with an out-of frame 1-bp deletion leads to a frameshift with translational read-through of the natural stop codon. The molecular mechanism is demonstrated by an in vitro translation study. The model of mutated cytochrome b5 reductase protein possessing 46 additional amino acids was obtained by homology modelling. The mutation causes an alteration of hydrophobicity in the carboxyl-terminal portion, resulting in the conformation being drastically disturbed by the presence of 46 supplementary amino acids. The identical mutation was found in the heterozygous state in the patient's parents and sister. Identification of this new mutation enabled us to perform the molecular prenatal diagnosis of type II RCM at the DNA level. PMID- 15813913 TI - Band 3Tambau: a de novo mutation in the AE1 gene associated with hereditary spherocytosis. Implications for anion exchange and insertion into the red blood cell membrane. AB - Hereditary spherocytosis (HS) is attributed to red blood cell membrane protein defects, caused by mutations in ankyrin, spectrin, band 3 and protein 4.2. In this study, the presence of band 3 mutations was investigated in a patient presenting mild HS and band 3 deficiency. Using single strand conformation polymorphism analysis, a shift in exon 16 of the band 3 gene was found. DNA sequencing revealed a point mutation 2102 T>C, changing methionine at position 663 to lysine. The M663K substitution was not found in either the parents or in the siblings, and the restriction fragment length polymorphism analysis of 100 alleles from a random Brazilian population did not reveal this mutation, suggesting that this gene defect is more likely to be a de novo mutation, causing HS. Flow cytometry of eosin-5-isothiocyanate (EITC)-labelled erythrocytes showed, in the patient, 54% of band 3 protein content vs. 78% based on the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, suggesting that flow cytometry is a more sensitive method and may be used as a diagnostic tool in membrane disorders related to band 3 deficiency. The characterisation of novel AE1 mutations is helpful to improve the understanding of the role of band 3 protein in cell physiology. PMID- 15813914 TI - Autologous stem cell transplantation in patients with high-risk plasmacytoma. AB - Although autologous stem cell transplantation (ASCT) is considered standard treatment in patients with multiple myeloma (MM), limited experience is available on this approach in patients with plasmacytoma (PC). Twelve patients with high risk PC received ASCT in Finland 1994-2002. There were nine males and three females with a median age of 50 yr (32-64). Ten patients had a PC of bone, whereas two patients had extramedullary PCs. The median time from the diagnosis to ASCT was 9 months (5-100). At the time of ASCT six patients were in first complete remission (CR) or partial remission (PR), in four patients the disease was refractory to the first line therapy and two patients had relapsed. High-dose therapy consisted of melphalan (MEL)200 (n = 7), MEL200 x 2 (n = 3) or total body irradiation (TBI)-MEL140 (n = 2). No transplant-related deaths occurred. After ASCT eight patients (67%) were in CR, one patient in very good PR and one patient in PR; two patients were non-responders. With a median follow-up of 48 months from ASCT, 11 patients (92%) are alive. Six patients (50%) have relapsed or progressed 3-81 months from ASCT. ASCT is feasible in this patient population resulting in promising overall survival. A randomised trial is needed to assess the real value of ASCT when compared with other treatment options in patients with high-risk PC. PMID- 15813915 TI - In vitro cytotoxic effect of proteasome inhibitor bortezomib in combination with purine nucleoside analogues on chronic lymphocytic leukaemia cells. AB - OBJECTIVE: The anti-tumour in vitro activity of proteasome inhibitor bortezomib (PS-341, VELCADE) in combination with purine nucleoside analogues, cladribine (2 CdA) and fludarabine (FA) was tested in lymphocytes derived from 26 patients with B-cell chronic lymphocytic leukaemia (B-CLL). METHODS: Cell viability was assessed by propidium iodide staining, and apoptosis by annexin-V and caspase activation flow cytometry assays. Additionally, expression of the apoptosis regulating proteins Bax, Bak, Bid, Bcl-w, Bcl-2, XIAP and Mcl-1 was evaluated in B-CLL lymphocytes. RESULTS: Bortezomib alone induced significant, dose-dependent cytotoxicity starting from the low concentration 2.5 nm, inducing apoptosis of B CLL cells. Combination of this agent with 2-CdA or FA resulted in an increase of cytotoxicity when compared with that mediated by single drugs. The observed increase was especially evident when 5 nm of bortezomib were combined with suboptimal doses of 2-CdA or FA. The combination index (CI) was 0.87 for bortezomib + 2-CdA and 0.82 for bortezomib + FA, indicating an evident additive effect of these combinations. Moreover, B-CLL cells were more sensitive to proteasome inhibitor used alone or combined with 2-CdA or FA comparing to CD3+ lymphocytes. Corresponding to enhanced apoptosis, the expression levels of several apoptosis-regulating proteins were altered. The most pronounced changes were down-regulation of XIAP and up-regulation of Bid proteins by the combination of bortezomib with either 2-CdA or FA. CONCLUSIONS: This study suggest that the in vitro cytotoxic effect through proteasome inhibition by bortezomib can be increased substantially with low doses of the purine nucleoside analogues, 2-CdA and FA, and that this effect on B-CLL cell is selectively higher than on normal, CD3-positive lymphocytes. PMID- 15813916 TI - Long-term outcomes for unselected patients with acute myeloid leukemia categorized according to the World Health Organization classification: a single center experience. AB - The actual utility of a new classification system of acute myeloid leukemia (AML) recently introduced by the World Health Organization (WHO) has not been thoroughly investigated yet. In this study, we evaluated long-term outcomes of unselected AML patients categorized according to the new WHO classification. Between 1990 and 2002, 109 adult AML cases were referred to our hospital. For the entire population, the median survival duration was 1.2 yr with a 5-yr survival rate of 31%. AML with recurrent genetic abnormalities accounted for 26%, AML with multilineage dysplasia for 29%, therapy-related AML for 13%, and AML not otherwise categorized for 32% of classifiable cases. Among the four groups, a significant difference was observed in terms of overall survival (P < 0.0001). Univariate analysis showed that six variables affected survival: cytogenetic risk, age, multilineage dysplasia, prior chemo/radiotherapy, type of treatment (intensive or palliative), and transplantation. However, in multivariate analysis no adverse prognostic impact of multilineage dysplasia and prior chemo/radiotherapy was detected (P = 0.4979 and 0.8702), whereas cytogenetic risk and patient age maintained their prognostic value (P = 0.0005 and 0.0100). These results indicate that outcomes for AML patients appear to be distinguished on the basis of the WHO classification, but the prognostic significance of multilineage dysplasia and prior therapy is lost after adjusting for cytogenetic risk and age. Our findings suggest that the WHO classification may be strengthened by greater emphasis on genetic/cytogenetic information. PMID- 15813917 TI - Methylation status analysis of cell cycle regulatory genes (p16INK4A, p15INK4B, p21Waf1/Cip1, p27Kip1 and p73) in natural killer cell disorders. AB - Natural killer (NK) cell disorders are rare diseases. Genetic abnormalities of the several tumor suppressor genes, including p15INK4B, p16INK4A/p14ARF, p53, p73, and Rb genes have been reported. Deletions and point mutations of these genes are frequently detected in these diseases. It has been reported that tumor suppressor genes are inactivated by DNA methylation of the promoter region and/or first exon of the genes in a variety of human cancers. In this study we analyze the methylation status of the genes associated with cell cycle regulation, including p16INK4A, p15INK4B, p21/Waf1/Cip1, p27/Kip1, p73, and p14ARF, by methylation specific (MS) PCR and/or bisulfite sequencing. We examined 29 cases of NK cell disorders (five aggressive NK cell leukemia/lymphoma, three blastic NK cell lymphoma/leukemia, five nasal NK cell lymphoma, three myeloid/NK cell precursor acute leukemia, 13 chronic NK lymphocytosis). We found methylation of the first exon of the p16INK4A gene in two cases (one aggressive, one blastic), and methylation of the p14ARF gene in one aggressive NK cell leukemia. Bisulfite sequencing revealed that methylation of the p15 and p27 genes was rare in these disorders. MS-PCR suggested that the p73 and p21 genes were methylated in seven cases, respectively (p73: one blastic, one nasal, five chronic; p21: one myeloid/NK, one aggressive, one nasal, and four chronic); bisulfite sequencing confirmed that methylated alleles of these genes were dominant in the samples except three cases (one myeloid/NK, one aggressive, and one chronic) in which methylated alleles of the p21 genes were less than 34% of all alleles. These results suggested that inactivation of the cell cycle regulatory genes by DNA methylation could be associated with tumorigenesis in NK cell disorders, not only aggressive subtypes but also chronic subtype. PMID- 15813918 TI - Comparison of DXA, QCT and trabecular structure in beta-thalassaemia. AB - Osteopathy, as a major feature of homozygous beta-thalassaemia, is a multifactorial disorder, not fully understood. We studied the lumbar vertebrae of 48 patients using Dual-Energy X-ray Absorptiometry (DXA) and Quantitative Computed Tomography (QCT), and we focused on structural properties, assessed by High Resolution Computed Tomography (HRCT). Bone Mineral Density (BMD) values were expressed as Z-scores and the results were correlated. The effect of age, sex, and type of thalassaemia and hormonal factors on BMD was assessed. We estimated, with HRCT, the cortex integrity and the number and thickness of trabeculae; the latter were classified to a three-grade scale. Our results showed the overall prevalence of osteoporosis to be 44% with DXA and 6% with QCT. Both techniques revealed an inverse correlation between age and BMD, whereas hormonal factors demonstrated associations with QCT and DXA measurements. The correlation coefficient between DXA's BMD and QCT's trabecular BMD was 0.545 (P < 0.001) whereas the corresponding value for Z-scores was r = 0.491 (P < 0.001). The classification of the patients into normal, osteopenic and osteoporotic categories, using QCT's Z, was in better agreement with the assignment based on trabecular number (K = 0.209, P = 0.053) than the classification using DXA's Z (K = 0.145, P = 0.120). Cortex evaluation by HRCT showed discontinuity in 15 patients. Both methods indicate a progression of osteoporosis with age. Hormonal deficiency is associated with thalassaemic osteoporosis whereas the visual estimation of cortex indicates that Thalassaemia Intermedia (TI) patients could be more affected than Thalassaemia Major (TM). Using the trabecular number as an indicator of osteoporosis, it seems that QCT may evaluate osteopathy better than DXA. Since the former has the ability to measure trabecular and cortical BMD separately, it could give early indication of which changes more rapidly and to what degree. PMID- 15813919 TI - Successful treatment of disseminated aspergillosis with the combination of voriconazole, caspofungin, granulocyte transfusions, and surgery followed by allogeneic blood stem cell transplantation in a patient with primary failure of an autologous stem cell graft. AB - The treatment of disseminated aspergillus infections in neutropenic patients remains a major challenge in spite of several new antifungal drugs. We report the case of a patient with multiple myeloma in prolonged neutropenia after primary failure of an autologous stem cell graft who developed invasive aspergillosis despite voriconazole monotherapy. He responded to a combination of voriconazole and caspofungin, supported by granulocyte transfusions and surgery. A subsequent allogeneic peripheral blood stem cell transplantation did not lead to recurring aspergillus infection. The patient is well and free of clinical disease with respect to the fungal infection and myeloma more than 18 months after the allogeneic transplantation. PMID- 15813920 TI - Use of rituximab to treat refractory Diamond-Blackfan anemia. AB - We report here the first case with Diamond-Blackfan anemia (DBA) who responded to rituximab. The patient is an 8-yr-old Japanese girl with refractory DBA accompanied by complex congenital heart disease. She received two doses of rituximab, 375 mg/m(2)/wk. She became transfusion independent 6 months after the treatment without any serious side effect. However, after 8 months of transfusion free period, her condition returned to the pretreatment level with recovery of peripheral B cells. Rituximab may be a successful therapy for refractory DBA where B cells play a crucial role in the pathogenesis of the severe anemia. PMID- 15813921 TI - Stable remission after administration of rituximab in a patient with primary hepatic marginal zone B-cell lymphoma. AB - We describe a case of primary hepatic marginal zone B-cell lymphoma in a 36-year old Caucasian male with a history of chronic hepatitis B infection. Immunohistochemically, extensive infiltration by a CD20-positive, CD5- negative and CD10-negative lymphoid cell population displaying a follicular arrangement was detected. Molecular analysis of immunoglobulin heavy chain gene rearrangements confirmed the clonal expansion of lymphoma cells. Fourteen months after surgical treatment, the tumour recurred in close proximity to the liver hilus, hampering further surgery. Therefore, we implemented a therapy using the monoclonal anti-CD20-antibody rituximab in a dose of 375 mg/m(2), administered four times once a week. Six, 10, 18, and 26 months later the recurrent lymphoma could no longer be detected as shown by abdominal ultrasonography and CT. This case report demonstrates the difficulties of treating this extremely rare liver disease and shows its response to rituximab therapy. PMID- 15813922 TI - Relapse of autoimmune thrombocytopenia following a positive skin prick test. PMID- 15813923 TI - Is CD20 the only target available for radionuclide therapy in lymphoproliferative disorders? PMID- 15813924 TI - Regulation of Homer and group I metabotropic glutamate receptors by nicotine. AB - The present study focuses on the nicotine-induced modulation of mRNA and protein expression of a number of genes involved in glutamatergic synaptic transmission in rat brain over different time periods of exposure. A subchronic (3 days) but not the chronic (7 or 14 days) administration of nicotine resulted in the up regulation of Homer2a/b mRNA in the amygdala while in the ventral tegmental area (VTA) no change in expression of either Homer2a/b or Homer1b/c was observed. Although the increase in Homer2a/b mRNA was not translated into the protein level in the amygdala, a slight but significant up-regulation of Homer1b/c protein was observed in the same region at day 3. Both Homer forms were up-regulated at the protein level in the VTA at day 3. In the nucleus accumbens, 14 days of nicotine treatment up-regulated mRNA of Homer2b/c by 68.2% (P < 0.05), while the short form Homer1a gene was down-regulated by 65.0% at day 3 (P < 0.05). In regard to other components of the glutamatergic signalling, we identified an acute and intermittent increase in the mRNA and protein levels of mGluR1 and mGluR5 in the amygdala. In the VTA, however, the effects of nicotine on mGluR mRNA expression were long-lasting but rather specific to mGluR1. Nevertheless, mGluR1 protein levels in the VTA area were up-regulated only at day 3, as in the amygdala. These data provide further evidence for the involvement of nicotine in the glutamatergic neuronal synaptic activity in vivo, suggesting a role for the newly identified Homer proteins in this paradigm. PMID- 15813925 TI - Coronin 3 and its role in murine brain morphogenesis. AB - Coronins belong to the fundamental WD40-repeat proteins. They are mainly found at the submembraneous area, they bind F-actin in vitro, and most of the seven mammalian coronins have unclear roles. Coronin 3 is abundantly expressed in the adult CNS. All murine brain areas express coronin 3 during embryogenesis and the first postnatal stages. Expression in grey matter decreases postnatally, except for hippocampal pyramidal and dentate gyrus neurons, and cerebellar Purkinje cells, while levels in white matter increase in the course of myelination. Consistently, coronin 3 is abundant in differentiating neuro-2a and PC-12 cells and in primary oligodendrocytes. Treatment with PKC activator PMA reduced coronin 3 protein levels. To address its functions, neuro-2a and PC-12 cells were transfected with GFP-tagged coronin 3 versions. Full-length coronin 3 among other areas localized to outgrowing neurites, whereas truncated proteins efficiently suppressed neurite formation. Our results favour a role for coronin 3 in neuron morphogenesis and possibly migration. PMID- 15813926 TI - Neuronal and glial expression of the adhesion molecule TAG-1 is regulated after peripheral nerve lesion or central neurodegeneration of adult nervous system. AB - Expression of the cell adhesion molecule TAG-1 is down-regulated in adult brain, with the exception of certain areas exhibiting structural plasticity. Here, we present evidence that TAG-1 expression persists also in adult rat spinal cord and dorsal root ganglia (DRG), and can be up-regulated after injury. On Western blots of adult tissue, TAG-1 is detected as a 135-kDa band, with an additional specific 90-kDa band, not present in developing tissue. TAG-1 expression is found both in DRG neurons and in Schwann cells, particularly those associated with the peripherally projecting DRG processes. Quantitative in situ hybridization revealed that TAG-1 expression is significantly higher in small neurons that give rise to unmyelinated fibers, than in large DRG neurons. The regulation of TAG-1 was then examined in two different lesion paradigms. After a sciatic nerve lesion, TAG-1 expression is not up-regulated in DRG neurons, but decreases with time. At the lesion site, reactive Schwann cells up-regulate TAG-1, as demonstrated by both immunohistochemistry and in situ hybridization. In a second paradigm, we injected kainic acid into the spinal cord that kills neurons but spares glia and axons. TAG-1 is up-regulated in the spinal neuron-depleted area as well as in the corresponding dorsal and ventral roots, associated with both target-deprived afferent fibers and with the non-neuronal cells that invade the lesion site. These results demonstrate a local up-regulation of TAG-1 in the adult that is induced in response to injury, suggesting its involvement in axonal re-modelling, neuron-glia interactions, and glial cell migration. PMID- 15813927 TI - Indirect phosphorylation-dependent modulation of postsynaptic nicotinic acetylcholine responses by 5-hydroxytryptamine. AB - Ionotropic nicotinic acetylcholine (ACh) receptors have been shown to be modulated by protein kinase-mediated phosphorylation in vitro. Here we demonstrate that 5-hydroxytryptamine (5-HT) can downregulate postsynaptic nicotinic ACh responses, elicited in an identified arthropod motoneuron in situ, by a mechanism dependent on protein kinase activity. Serotonergic modulation can be mimicked by perfusion with membrane-permeable analogues of either adenine (cAMP) or guanine (cGMP) cyclic nucleotides, and is prolonged in the presence of phosphodiesterase inhibitors. Furthermore, suppression of the ACh response by 5 HT is blocked by specific competitive inhibitors of protein kinase A and G, as well as the broad specificity protein kinase inhibitor staurosporine. The protein phosphatase inhibitor cantharidin similarly blocks recovery of the ACh response from suppression mediated by 5-HT. Thus, it appears that the nicotinic ACh response is modulated by a cAMP-mediated phosphorylation-dependent intracellular signalling pathway that is distinct from the direct block of mammalian nicotinic ACh receptors by 5-HT previously reported in vitro. PMID- 15813928 TI - Differences between cation-chloride co-transporter functions in the visual cortex of pigmented and albino rats. AB - Albinism in mammals is accompanied by specific morphological and functional alterations of the visual system. To understand their cellular basis we studied the physiological characteristics and transmembrane currents of pyramidal neurons in 350-microm-thick slices of visual cortex from pigmented and albino rats using whole-cell and gramicidin perforated patch-clamp recordings. The resting membrane potential was significantly more positive and the rheobase was significantly lower in neurons of layers II/III and V in albinos as compared with pigmented rats. No significant differences were found in the input resistance, time constant and chronaxy. Whereas the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptor-mediated currents were not significantly different, the maximum gamma-aminobutyric acid (GABA)(A) receptor (GABA(A)R) mediated currents and miniature inhibitory postsynaptic currents showed significantly lower amplitudes in neurons of layer V in visual cortex of albinos as compared with pigmented rats. The reversal potential of the GABA(A)R-mediated currents (E(GABA)) was significantly shifted to more positive values in albinos. Pharmacological experiments showed that this shift could be caused by an increased action of the inward chloride co-transporter NKCC1 and reduced action of the outward chloride co-transporter KCC2 in albino rats. This difference seems to be restricted to the visual cortex because in pyramidal neurons from frontal cortex E(GABA) was not significantly different in albinos as compared with pigmented rats. These results are discussed in relation to functional alterations in the albino visual system. PMID- 15813930 TI - Differential regulation of type I and type II interleukin-1 receptors in focal brain inflammation. AB - Most pathologies of the brain have an inflammatory component, associated with the release of cytokines such as interleukin-1beta (IL-1beta) from resident and infiltrating cells. The IL-1 type I receptor (IL-1RI) initiates a signalling cascade but the type II receptor (IL-1RII) acts as a decoy receptor. Here we have investigated the expression of IL-1beta, IL-1RI and IL-1RII in distinct inflammatory lesions in the rat brain. IL-1beta was injected into the brain to generate an inflammatory lesion in the absence of neuronal cell death whereas neuronal death was specifically induced by the microinjection of N-methyl-D aspartate (NMDA). Using TaqMan RT-PCR and ELISA, we observed elevated de novo IL 1beta synthesis 2 h after the intracerebral microinjection of IL-1beta; this de novo IL-1beta remained elevated 24 h later. There was a concomitant increase in IL-1RI mRNA but a much greater increase in IL-1RII mRNA. Immunostaining revealed that IL-1RII was expressed on brain endothelial cells and on infiltrating neutrophils. In contrast, although IL-1beta and IL-1RI were elevated to similar levels in response to NMDA challenge, the response was delayed and IL-1RII mRNA expression was unchanged. The lesion-specific expression of IL-1 receptors suggests that the receptors are differentially regulated in a manner not directly related to the endogenous level of IL-1 in the CNS. PMID- 15813929 TI - Different responsiveness of striatonigral and striatopallidal neurons to L-DOPA after a subchronic intermittent L-DOPA treatment. AB - Early gene induction by L-DOPA in the striatum of dopamine denervated rats represents a useful way to study long-term modifications produced by this drug. The effects of acute and subchronic L-DOPA administration on zif-268 mRNA expression were compared in 6-hydroxydopamine-lesioned rats. Rats received a subchronic intermittent L-DOPA (6 mg/kg) treatment, which produces behavioural sensitization, a correlate of dyskinetic movements. Three days after interruption of subchronic treatment, zif-268 mRNA was evaluated after an L-DOPA challenge. Zif-268 mRNA levels increased in the lesioned dorsolateral striatum after either acute or subchronic L-DOPA administration. Double labelling of striatal cells with zif-268 and enkephalin or dynorphin mRNA probes was performed to assess neuronal activation in the indirect and direct output pathway. Single acute L DOPA significantly increased zif-268 in all striatal neurons reflecting a hyperresponsiveness of dopamine-depleted striatum. After subchronic L-DOPA, zif 268 mRNA labelling was still increased in the striatonigral pathway, limited to dynorphin(+) neurons, whereas in all other neurons it was similar to the control value. Results suggest that striatal neurons responding to acute L-DOPA differ from those responding to subchronic L-DOPA. L-DOPA-induced behavioural sensitization was associated to a down-regulation in the responsiveness of striatopallidal and striatonigral dynorphin(-) neurons, whereas in striatonigral neurons containing dynorphin a hyperresponsiveness to L-DOPA was observed. High levels of zif-268, together with a persistent hyperresponsiveness of striatonigral dymorphinergic neurons and hyporesponsiveness of striatopallidal neurons, by creating an unbalanced state of striatal efferent neurons, may be implicated in dyskinetic movements observed in Parkinson's disease (PD). PMID- 15813932 TI - Intracerebral study of gamma rhythm reactivity in the sensorimotor cortex. AB - The generators and functional correlates of gamma oscillations within the sensorimotor cortex remain unclear. With the goal of locating the oscillations' sources precisely and then studying the relationship between oscillatory reactivity and ongoing movement, we recorded stereoelectroencephalograms with intracerebral electrodes in eight epileptic subjects awaiting surgical treatment. The sensorimotor cortex was free of lesions and was exhaustively explored with the electrodes. Subjects were asked to perform various self-paced movements contralateral to the exploration zone, brief and sustained, distal movements and a pointing movement. We used the event-related desynchronization method to quantify the reactivity of the 40-60-Hz band before, during and after the performance of movement. A very focused, event-related synchronization of gamma rhythms was found in all subjects. It was predominantly observed in the primary sensorimotor area and its distribution was consistent with the functional map established using electrical stimulations. Two different temporal patterns were observed, the event-related synchronization of gamma rhythms was related either to movement onset or to movement offset but was never recorded before movement. This observation suggests that gamma oscillations are more probably related to movement execution than to motor planning. The different patterns argue in favour of multiple functional roles; it has been shown that gamma oscillations may support the efferent drive to the muscles and here we show that they are also likely to be related to somatosensory integration. We therefore suggest that gamma oscillations in the 40-60-Hz band may support afferent sensory feedback to the sensorimotor cortex during the performance of movement. PMID- 15813931 TI - Voltage-gated calcium channels play crucial roles in the glutamate-induced phase shifts of the rat suprachiasmatic circadian clock. AB - The resetting of the circadian clock based on photic cues delivered by the glutamatergic retinohypothalamic tract is an important process helping mammals to function adaptively to the daily light-dark cycle. To see if the photic resetting relies on voltage-gated Ca(2+) channels (VGCCs), we examined the effects of VGCC blockers on the glutamate-induced phase shifts of circadian firing activity rhythms of suprachiasmatic nucleus (SCN) neurons in hypothalamic slices. First, we found that a cocktail of amiloride, nimodipine and omega-conotoxin MVIIC (T-, L- and NPQ-type VGCC antagonists, respectively) completely blocked both phase delays and advances, which were, respectively, induced by glutamate application in early and late night. Next, we discovered that: (i) amiloride and another T type VGCC antagonist, mibefradil, completely obstructed the delays without affecting the advances; (ii) nimodipine completely blocked the advances while having less impact on delays; and (iii) omega-conotoxin MVIIC blocked largely, if not entirely, both delays and advances. Subsequent whole-cell recordings revealed that T-type Ca(2+) currents in neurons in the ventrolateral, not dorsomedial, region of the SCN were larger during early than late night, whereas L-type Ca(2+) currents did not differ from early to late night in both regions. These results indicate that VGCCs play important roles in glutamate-induced phase shifts, T type being more important for phase delays and L-type being so for phase advances. Moreover, the results point to the possibility that a nocturnal modulation of T-type Ca(2+) current in retinorecipient neurons is related to the differential involvement of T-type VGCC in phase delays and advances. PMID- 15813933 TI - Balanced inhibition and excitation underlies spike firing regularity in ventral cochlear nucleus chopper neurons. AB - Ventral cochlear nucleus stellate cells respond to characteristic frequency (CF) tones with sustained (C(S)), transient (C(T)) or onset chopping (O(C)) activity. The mechanisms underlying these different response patterns are not fully understood, and the present study used in vivo intracellular recordings (n = 42) in urethane-anaesthetized rats to examine the possible influence of inhibition on action potential regularity. Hyperpolarization following the offset of a CF tone burst was used as a measure of on-CF inhibition. A cluster analysis based on several membrane potential features, including on-CF inhibition, discriminated three groups in addition to the C(S) response type - two types of C(T) responses and the O(C) type. The different patterns of firing regularity exhibited by C(S/T) neurons reflected different thresholds or degrees of overlap between these cells' narrowly tuned excitatory and inhibitory inputs. C(T) cells with closely matched inhibitory and excitatory response areas showed substantial on-CF inhibition and the greatest decline in firing regularity during a CF tone, whereas those with a mismatch between their response areas showed lateral inhibition and a less marked decline in firing regularity. The presence of inhibition in C(S) neurons did not alter their firing regularity, possibly because of the lower threshold for excitation compared with inhibition. The latency, duration and frequency extent of sustained hyperpolarization in C(S/T) cells is inconsistent with the response properties of O(C) neurons, suggesting that another source(s) of inhibition influences firing regularity, and presumably response magnitude, in these neurons. PMID- 15813934 TI - Type grouping in skeletal muscles after experimental reinnervation: another explanation. AB - Type grouping signifies clustering of muscle fibres of the same metabolic type, and is a frequent finding in reinnervated muscles. To elucidate the mechanism behind it, the rat sciatic nerve was either autografted or grafted with hollow synthetic nerve grafts. Twelve weeks later the number and fibre area of the type I and type II muscle fibres in the gastrocnemic and anterior tibial muscles were determined after ATP-ase staining. The number and diameter of peroneal nerve fibres distal to the grafts were measured, and the number of Aalpha-nerve fibres was derived. Nearly all nerve and muscle morphometrical parameters changed equally in both experimental groups. However, type grouping occurred frequently only after autografting, whereas the number of nerve fibres and the number of Aalpha-nerve fibres increased in this group. Hence type grouping cannot be explained by increased intramuscular sprouting subsequent to a decrease in the number of innervating nerve fibres, as previously presumed. Regenerating axons branch along their course through the peripheral nerve. We propose that the probability of the occurrence of type grouping is related to the dispersion of sibling branches in the nerve. In the autograft, emerging branches are kept together by Schwann cell basal lamina scaffolds, in contrast to the hollow synthetic nerve grafts where the emerging branches become dispersed. Thus, in muscles reinnervated after autografting, the probability that nerve branches that arrive at a specific muscle territory are sibling branches is greater than after hollow tube grafting. Consequently, the probability that type grouping will occur is greater. PMID- 15813935 TI - Synaptic contacts between an identified type of ON cone bipolar cell and ganglion cells in the mouse retina. AB - We surveyed the potential contacts between an identified type of bipolar cell and retinal ganglion cells in the mouse. By crossing two existing mouse strains (line 357 and line GFP-M), we created a double transgenic strain in which GFP is expressed by all members of a single type of ON cone bipolar cell and a sparse, mixed population of retinal ganglion cells. The GFP-expressing bipolar cells appear to be those termed CB4a of Pignatelli & Strettoi [(2004) J. Comp. Neurol., 476, 254-266] and type 7 of Ghosh et al. [(2004) J. Comp. Neurol., 469, 70-82 and J. Comp. Neurol., 476, 202-203]. The labelled ganglion cells include examples of most or all types of ganglion cells present in the mouse. By studying the juxtaposition of their processes in three dimensions, we could learn which ganglion cell types are potential synaptic targets of the line 357 bipolar cell. Of 12 ganglion cell types observed, 10 types could be definitively ruled out as major synaptic targets of the line 357 bipolar cells. One type of monostratified ganglion cell and one bistratified cell tightly cofasciculate with axon terminals of the line 357 bipolar cells. Double labelling for kinesin II demonstrates colocalization of bipolar cell ribbons at the sites of contact between these two types of ganglion cell and the line 357 bipolar cells. PMID- 15813936 TI - Mechanisms of ATP action on motor nerve terminals at the frog neuromuscular junction. AB - We have shown previously that ATP inhibits transmitter release at the neuromuscular junction through the action on metabotropic P2Y receptors coupled to specific second messenger cascades. In the present study we recorded K(+) or Ca(2+) currents in motor nerve endings or blocked K(+) or Ca(2+) channels in order to explore the nature of downstream presynaptic effectors. Endplate currents were presynaptically depressed by ATP. Blockers of Ca(2+)-activated K(+) channels, such as iberiotoxin, apamin or tetraethylammonium, did not change the depressant action of ATP. By contrast, K(+) channel blocker 4-aminopyridine (4 AP) and raised extracellular Ca(2+) attenuated the effect of ATP. However, these effects of 4-AP and high Ca(2+) were reversed by Mg(2+), suggesting Ca(2+) dependence of the ATP action. Ba(2+) promoted the depressant action of ATP as did glibenclamide, a blocker of ATP-sensitive K(+) channels, or mild depolarization produced by 7.5 mm K(+). None of the K(+) channel blockers affected the depressant action of adenosine. Focal recording revealed that neither ATP nor adenosine affected the fast K(+) currents of the motor nerve endings. However, unlike adenosine, ATP or UTP, an agonist of P2Y receptors, reversibly reduced the presynaptic Ca(2+)-current. This effect was abolished by suramin, an antagonist of P2 receptors. Depressant effect of ATP on the endplate and Ca(2+)-currents was mimicked by arachidonate, which precluded the action of ATP. ATP reduced acetylcholine release triggered by ionomycin or sucrose, suggesting inhibition of release machinery. Thus, the presynaptic depressant action of ATP is mediated by inhibition of Ca(2+) channels and by mechanism acting downstream of Ca(2+) entry. PMID- 15813937 TI - mGluRs induce a long-term depression in the ventral tegmental area that involves a switch of the subunit composition of AMPA receptors. AB - Excitatory glutamatergic synapses on dopamine (DA) neurons of the ventral tegmental area (VTA) undergo long-lasting changes during conditioning of natural rewards and in response to drug exposure. It has been suggested that the ensuing context-dependent behavioural changes are associated with an increased efficacy of synaptic afferents determined by the balance of long-term potentiation (LTP) and long-term depression (LTD). However, the molecular nature of the forms of LTP/LTD involved remains elusive. Here, using acute rat brain slices, we describe a form of long-term depression (LTD) that was engaged by synaptic activity or exogenous agonists activating group I metabotropic glutamate receptors (mGluR) and was sensitive to mGluR1 antagonists. Prior to mGluR-LTD, AMPAR mediated excitatory postsynaptic currents (EPSCs) showed strong rectification at positive potentials and were sensitive to Joro spider toxin (JST), a selective blocker of GluR2-lacking AMPARs. After mGluR-LTD, AMPAR EPSCs had linear current-voltage relations and became insensitive to JST. We conclude that activation of mGluR1s triggers a redistribution exchanging native receptors for GluR2 containing AMPARs, ultimately causing LTD that may oppose pathological neuroadaptation. PMID- 15813938 TI - The novel cytosolic RING finger protein dactylidin is up-regulated in brains of patients with Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by a progressive degeneration of neurons along with deposition of amyloid plaques and the formation of neurofibrillary tangles. Neurodegeneration in AD follows both a spatial pattern of selective vulnerability and temporal staging of affected neurons. In order to address transcriptional changes associated with this selective vulnerability, we used subtractive hybridization of transcripts derived from human frontal cortex, which degenerates in late stages of AD, against transcripts of the inferior temporal cortex, which is affected both heavily and early in the course of AD. Moreover, we compared these to brain sections obtained from age-matched control subjects. We isolated a differentially expressed novel gene encoding a polypeptide that contained an amino-terminal C3HC4 RING finger domain, called dactylidin. It is ubiquitously expressed in all tissues examined and in situ hybridization of mouse brain sections revealed specific expression in neurons. Further, heterologous expression studies revealed a cytoplasmic localization of dactylidin and as all known cytoplasmic RING finger proteins function as ubiquitin protein ligases, an E3-like ligase function of dactylidin is probable. However, the up-regulation of dactylidin in highly vulnerable brain tissues of AD patients was confirmed by a quantitative PCR approach, suggesting that dactylidin may function early in the progression of neurodegenerative diseases. PMID- 15813939 TI - Short-term treatment with the antidepressant fluoxetine triggers pyramidal dendritic spine synapse formation in rat hippocampus. AB - The pathomechanism of major depressive disorder and the neurobiological basis of antidepressant therapy are still largely unknown. It has been proposed that disturbed hippocampal activity could underlie some of the cognitive and vegetative symptoms of depression, at least in part because of loss of pyramidal cell synaptic contacts, a process that is likely to be reversed by antidepressant treatment. Here we provide evidence that daily administration of the antidepressant fluoxetine to ovariectomized female rats for 5 days induces a robust increase in pyramidal cell dendritic spine synapse density in the hippocampal CA1 field, with similar changes appearing in CA3 after 2 weeks of treatment. This rapid synaptic remodelling might represent an early step in the fluoxetine-induced cascade of responses that spread across the entire hippocampal circuitry, leading to the restoration of normal function in the hippocampus. Hippocampal synaptic remodelling might provide a potential mechanism to explain certain aspects of antidepressant therapy and mood disorders, especially those associated with changes in reproductive state in women, that cannot be reconciled adequately with current theories for depression. PMID- 15813940 TI - Chronic stress in the adult dentate gyrus reduces cell proliferation near the vasculature and VEGF and Flk-1 protein expression. AB - Recent evidence has shown that cell proliferation in the adult hippocampal dentate gyrus occurs in tight clusters located near the vasculature. Also, changes in neurogenesis often appear parallel to changes in angiogenesis. Moreover, both these processes share similar modulating factors, like vascular endothelial growth factor (VEGF) and its receptor Flk-1. In an earlier study we found that chronic stress decreased new cell proliferation in the adult dentate gyrus. We here questioned whether these effects of chronic stress are mediated through the vasculature and whether they involve an angiogenic-signaling pathway. We therefore measured the surface area covered by the vasculature, the proportion of vascular-associated newborn cells, and analysed VEGF and Flk-1 protein expression in the hippocampus of a control, chronically stressed and recovery group of rats. Our results show that 32% of the proliferating cells in the rat hippocampus is vascular associated. Chronic stress affected this population of newborn cells to a significantly larger extent than the non-associated cells. Interestingly, after 3 weeks of recovery, the decreased proliferation not associated with the vasculature was more effectively restored than vascular associated proportion of proliferating cells. VEGF protein was expressed in high densities in GFAP-positive astrocytes located in the hilus, with VEGF-positive end feet extending into and often contacting the granule cells. After chronic stress, both VEGF and Flk-1 protein levels were significantly decreased in the granular cell layer, and again recovered after 3 weeks. This demonstrates that changes in angiogenic factors are implicated in the decreased adult proliferation found after chronic stress. PMID- 15813941 TI - Defective control and adaptation of reflex eye movements in mutant mice deficient in either the glutamate receptor delta2 subunit or Purkinje cells. AB - The ionotropic glutamate receptor delta2 subunit (GluRdelta2) is selectively expressed in cerebellar Purkinje cells and is implicated in long-term depression, synaptic formation and elimination. To study the effect of GluRdelta2 deficiency on motor control, we measured the vestibulo-ocular reflex (VOR) and optokinetic response (OKR) induced by sinusoidal rotation of the animal and/or the surrounding screen in two GluRdelta2 mutant mice: a GluRdelta2 knockout mouse (delta2-/-) and a lurcher mouse with a point mutation in the GluRdelta2 gene resulting in loss of all Purkinje cells. delta2-/- showed significantly higher VOR gain in the dark (VORD) than in the wild-type. In delta2-/-, the VOR gain in light was lower than that in the dark. The phase of OKR lagged more in delta2-/- than in lurcher and wild-type mice. Both mutant mice failed to change the VORD or OKR gain adaptively in response to sustained vestibular and/or visual stimulation. Basal properties of VOR and OKR changed little by lesion of the flocculus, but they changed substantially by lesion of the inferior olivary nuclei (IO). The abnormal VOR gain and OKR phase delay were clearly reduced in delta2-/- by the latter lesion. Our results indicate that failures in the GluRdelta2-dependent synaptic regulation affect motor performance more severely than loss of cerebellar cortical outputs. This study suggests that the anomalies in delta2-/- are dependent on inputs from IO and that GluRdelta2 deficiency changed properties of not only the cerebellar cortex but also the brainstem neuronal pathways controlling reflex eye movements during development. PMID- 15813942 TI - Behavioural phenotypes of hypomorphic KCC2-deficient mice. AB - Hyperpolarizing fast inhibitory neurotransmission by gamma-aminobutyric acid and glycine requires an efficient chloride extrusion mechanism in postsynaptic neurons. A major effector of this task in adult animals is the potassium-chloride co-transporter KCC2 that is selectively and abundantly expressed postsynaptically in most CNS neurons. Yet, the role of KCC2 in adult brain at the systems level is poorly known. Here, we characterize the behaviour of mice doubly heterozygous for KCC2 null and hypomorphic alleles that retain 15-20% of normal KCC2 protein levels in the brain. These hypomorphic KCC2-deficient mice were viable and fertile but weighed 15-20% less than wild-type littermates at 2 weeks old and thereafter. The mice displayed increased anxiety-like behaviour in several tests including elevated plus-maze and were more susceptible to pentylenetetrazole induced seizures. Moreover, the mice were impaired in water maze learning and showed reduced sensitivity to tactile and noxious thermal stimuli in von Frey hairs, hot plate and tail flick tests. In contrast, the mice showed normal spontaneous locomotor activity in open field and Y-maze tests, and intact motor coordination in rotarod and beam tests. The results suggest that requirements for KCC2-dependent fast hyperpolarizing inhibition may differ among various functional systems of the CNS. As shunting inhibition is expected to be intact in KCC2-deficient neurons, these mice may provide a useful tool to study the specific functions and relative importance of hyperpolarizing fast synaptic inhibition in adult CNS that may have implications for human neuropsychiatric disorders, such as epilepsy, pain and anxiety. PMID- 15813943 TI - Serotonin refines the locomotor-related alternations in the in vitro neonatal rat spinal cord. AB - Serotonergic projections from raphe nuclei arrive in the lumbar enlargement of the spinal cord during the late fetal period in the rat, a time window during which the locomotor-related left/right and flexor/extensor coordinations switch from synchrony to alternation. The goal of the present study was to investigate the role played by serotonin (5-HT) in modulating the left/right and flexor/extensor alternations. Fictive locomotion was induced by bath application of N-methyl-D,L-aspartate (NMA) in the in vitro neonatal rat spinal cord preparation. By means of cross-correlation analysis we demonstrate that 5-HT, when added to NMA, improves left/right and flexor/extensor (recorded from the 3rd and 5th lumbar ventral roots, respectively) alternations. This effect was partly reproduced by activation of 5-HT(2A/2C) receptors. We then tested the contribution of endogenous 5-HT to NMA-induced fictive locomotion. Reducing the functional importance of endogenous 5-HT, either by inhibiting its synthesis with daily injections of p-chloro-phenylalanine (PCPA), starting on the day of birth, or by application of ketanserin (a 5-HT(2) receptor antagonist) or SB269970 (a 5 HT(7) receptor antagonist), disorganized the NMA-induced locomotor pattern. This pattern was restored in PCPA-treated animals by adding 5-HT to the bath. Blocking 5-HT(7) receptors disorganized the locomotor-like rhythm even in the absence of electrical activity in the brain stem, suggesting that NMA applied to the spinal cord does not cause 5-HT release by activating a spino-raphe-spinal loop. These results demonstrate that 5-HT is critical in improving the locomotor-related alternations in the neonatal rat. PMID- 15813944 TI - A single time-window for protein synthesis-dependent long-term memory formation after one-trial appetitive conditioning. AB - Protein synthesis is generally held to be essential for long-term memory formation. Often two periods of sensitivity to blockade of protein synthesis have been described, one immediately after training and another several hours later. We wished to relate the timing of protein synthesis-dependence of behavioural long-term memory (LTM) formation to an electrophysiological correlate of the LTM memory trace. We used the snail Lymnaea because one-trial appetitive conditioning of feeding using a chemical conditioned stimulus leads to a stable LTM trace that can be monitored behaviourally and then electrophysiologically in preparations made from the same animals. Anisomycin (an inhibitor of translation) injected 10 min after training blocked behavioural LTM formation. Actinomycin D (an inhibitor of transcription) was also effective at 10 min. When anisomycin, at doses shown to be effective in blocking central nervous system protein synthesis, was injected at 1, 2, 3, 4, 5 and 6 h after training there was no effect on recall. These results indicate that there is a single period of sensitivity to protein synthesis inhibition in Lymnaea lasting for between 10 min and 1 h after training with no evidence for a second window of sensitivity. An electrophysiological correlate of LTM was found to be sensitive to anisomycin injected 10 min after training. It is unusual to find only one period of protein synthesis-dependence in detailed time-course studies of LTM, and this suggests that the consolidation processes involving protein synthesis are relatively rapid in one-trial appetitive conditioning and complete within 1 h of training. PMID- 15813945 TI - Impairment of conditioned freezing to tone, but not to context, in Fyn-transgenic mice: relationship to NMDA receptor subunit 2B function. AB - We previously demonstrated that transgenic mice overexpressing Fyn tyrosine kinase exhibit higher seizure susceptibility and enhanced tyrosine phosphorylation of several proteins, including the N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B). In the present study, we analysed behavioural phenotypes, especially conditioned fear responses, of Fyn-transgenic (TG) mice to better understand the role of Fyn in learned emotional behaviour. Tone-dependent conditioned freezing was significantly attenuated in Fyn-TG mice, whereas context dependent freezing was unaffected. Neither massed nor spaced conditioning ameliorated the attenuation of tone-dependent freezing. However, the selective NR2B antagonist ifenprodil, when administered before conditioning, restored tone dependent freezing in Fyn-TG mice at a dose that did not affect freezing in wild type (WT) mice. These results suggest that impairment of tone-dependent conditioned freezing in Fyn-TG mice is caused by disruption of the NR2B containing NMDA receptor function. Tyrosine phosphorylation of brain proteins, including NR2B, was enhanced in Fyn-TG mice compared with that in WT mice. We also found that ifenprodil significantly suppressed the enhanced tyrosine phosphorylation. Thus, our data support the notion that NMDA receptor activity is tightly correlated with protein tyrosine phosphorylation, and Fyn might be one key molecule that controls tone-dependent conditioned freezing through the regulation of NMDA receptor function. PMID- 15813946 TI - Changes in signaling pathways regulating neuroplasticity induced by neurokinin 1 receptor knockout. AB - Neurokinin 1 (NK-1) receptor knockout mice showed behavioral responses similar to animals chronically treated with antidepressants. The aim of this study was to analyse, in NK-1 receptor knockout, the molecular modifications of signaling pathways involved in the pathophysiology of depression and antidepressant mechanism. We found, in total cell cytosol from the prefrontal/frontal cortex, hippocampus and striatum, a marked up-regulation of Ca(2+)-independent enzymatic activity and Thr(286) autophosphorylation of Ca(2+)/calmodulin-dependent protein kinase (CaMK) II. Similar changes in CaMKII regulation were previously observed in rats chronically treated with antidepressants. In striatum, up-regulation of the activity and phosphorylation of CaMKII was also found in the homogenate and synaptosomes. No major changes were observed in the Ca(2+)-dependent kinase activity, with the exception of homogenate from the prefrontal/frontal cortex. We also analysed the expression and phosphorylation of presynaptic proteins, which modulate synaptic vesicle trafficking and exocytosis, and found a marked decrease in synapsin I total expression and basal phosphorylation of Ser(603) (the phosphorylation site for CaMKII) in the prefrontal/frontal cortex. Accordingly, the Ca(2+)/calmodulin-dependent posthoc endogenous phosphorylation of synapsin I in the same area was increased. The knockout of NK-1 receptor had no consequences on the expression or phosphorylation levels of the transcription factor cAMP responsive element-binding protein and its regulating kinase CaMKIV. However, phosphorylation of ERK1/2-mitogen-activated protein kinases was reduced in the hippocampus and striatum, again resembling an effect previously observed in antidepressant-treated rats. These results show similarities between NK-1 knockouts and animals chronically treated with antidepressants and support the putative antidepressant activity of NK-1 receptor antagonists. PMID- 15813947 TI - Endogenous enkephalins, not endorphins, modulate basal hedonic state in mice. AB - The aversive response to naloxone administration observed in human and animal studies suggests the presence of an endogenous opioid tone regulating hedonic state but the class(es) of opioid peptides mediating such opioid hedonic tone is uncertain. We sought to address this question using mice deficient in either beta endorphin or pro-enkephalin in a naloxone-conditioned place aversion paradigm. Mice received saline in the morning in one chamber and either saline or naloxone (0.1, 1 or 10 mg/kg, s.c.) in the afternoon in another chamber, each day for 3 days. On the test day they were given free access to the testing chambers in the afternoon and the time spent in each chamber was recorded. Whereas wild-type and beta-endorphin-deficient mice exhibited a robust conditioned place aversion to naloxone, pro-enkephalin knockout mice failed to show aversion to naloxone at any dose tested. In contrast, these mice showed a normal conditioned aversion to the kappa opioid receptor agonist, U50,488 (5 mg/kg), and to LiCl (100 mg/kg) indicating that these mice are capable of associative learning. In a separate experiment, pro-enkephalin knockout mice, similar to wild-type and beta-endorphin deficient mice, demonstrated a significant conditioned place preference to morphine (2.5, 5 and 10 mg/kg s.c.). These data suggest that enkephalins, but not endorphins, may mediate an endogenous opioid component of basal affective state and also indicate that release of neither endogenous enkephalins nor endorphins is critical for the acquisition or expression of the association between contextual cues and the rewarding effect of exogenously administered opiates. PMID- 15813948 TI - Altered Fos expression in neural pathways underlying cue-elicited drug seeking in the rat. AB - Cocaine treatment paired with environmental cues establishes a conditioned place preference for that environment. Following expression of this preference, rats show elevated levels of immediate early genes (e.g. c-fos) in the prelimbic cortex (PrL), basolateral amygdala complex (BLC) and nucleus accumbens core (NAcc) compared to drug-unpaired controls. The PrL and BLC are reciprocally connected and both project to the NAcc. Together with the immediate early gene findings, these connections suggest the regions interact as a circuit contributing to cue-elicited drug seeking. To study this circuit, we iontophoresed Fluorogold (FG) into one brain region and assessed colocalization of FG with place preference-induced Fos in the others. Following FG iontophoresis in either the PrL or NAcc, more BLC cells double-labelled for Fos and FG were found in drug-paired than unpaired animals. Following FG iontophoresis in either the BLC or NAcc, no differences were found in the absolute number of PrL Fos/FG cells. This pattern of colocalization suggests that exposure to cocaine associated cues leads to greater activation of the BLC's efferents to both the PrL and NAcc, while PrL output to the NAcc and BLC is unaffected in IEG expression. These results complement recent findings that suggested attenuated PrL output during place preference expression. Our findings support the view that the BLC, rather than the PrL, provides significant excitatory driving to the NAcc during cue-elicited drug seeking. PMID- 15813949 TI - Stimulation through electrodes implanted near the subthalamic nucleus activates projections to motor areas of cerebral cortex in patients with Parkinson's disease. AB - High-frequency electrical stimulation through electrodes implanted in the subthalamic nucleus (STN) has been shown to reduce significantly the cardinal symptoms of Parkinson's disease (PD). Despite the success of this treatment, the mechanisms of action of stimulation are poorly understood. To elucidate further the mechanisms of action of deep brain stimulation and its effects on cortical activity, we recorded electroencephalographic potentials from 61 scalp-surface electrodes during low-frequency (5-10 Hz) bipolar stimulation in 11 patients with advanced PD (14 implanted electrodes were tested). In all electrodes tested, stimulation through at least one of the four contacts produced a medium-latency waveform with an average onset of 14 +/- 3 ms and peak at 23 +/- 4 ms. This potential typically increased in magnitude across contacts from ventral to dorsal. Within-subject comparisons of median nerve somatosensory evoked potentials demonstrated that the generator of the medium-latency potential was within the primary sensorimotor cortex or lateral premotor cortex ipsilateral to stimulation. The timing and topography of this potential were consistent with indirect activation of the cortex by excitation of pallido-thalamic axons that traverse the dorsal aspect of the STN. The potential evoked by stimulation through the contact used for optimal clinical effect was highly variable across electrodes and frequently different from the medium-latency potential described above, suggesting that the neuronal elements mediating the medium-latency potential were different from those that mediate the clinical effects. PMID- 15813950 TI - Levodopa-induced modulation of subthalamic beta oscillations during self-paced movements in patients with Parkinson's disease. AB - Excessive synchronization of neural activity in the beta frequency band ( approximately 20 Hz) within basal ganglia circuits might contribute to the paucity and slowness of movement in Parkinson's disease (PD). Treatment with dopaminergic drugs reduces the background level of beta frequency band synchronization in the subthalamic nucleus (STN), but has not been shown to increase the proportion of beta activity that is suppressed before voluntary movement in PD. We assessed changes in the event-related desynchronization (ERD) in the beta frequency band of local field potential signals from the region of the STN in 14 patients with PD as they performed self-paced movements of a joystick before and after levodopa administration. The dopamine precursor, levodopa, increased the duration and magnitude of the premovement beta ERD, but did not alter postmovement synchronization in the beta band. Both the latency and magnitude of the beta ERD inversely correlated with the degree of motor impairment. These findings suggest that the beta ERD recorded in the STN area reflects motor-preparative processes that are at least partly dependent on dopaminergic activity within the basal ganglia. PMID- 15813951 TI - Dopamine depletion increases the power and coherence of beta-oscillations in the cerebral cortex and subthalamic nucleus of the awake rat. AB - Local field potentials (LFPs) recorded from the subthalamic nucleus (STN) of untreated patients implanted with stimulation electrodes for the treatment of Parkinson's disease (PD) demonstrate strong coherence with the cortical electroencephalogram over the beta-frequency range (15-30 Hz). However, studies in animal models of PD emphasize increased temporal coupling in cortico-basal ganglia circuits at substantially lower frequencies, undermining the potential usefulness of these models. Here we show that 6-hydroxydopamine (6-OHDA) lesions of midbrain dopamine neurons are associated with significant increases in the power and coherence of beta-frequency oscillatory activity present in LFPs recorded from frontal cortex and STN of awake rats, as compared with the healthy animal. Thus, the pattern of synchronization between population activity in the STN and cortex in the 6-OHDA-lesioned rodent model of PD closely parallels that seen in the parkinsonian human. The peak frequency of coherent activity in the beta-frequency range was increased in lesioned animals during periods of spontaneous and sustained movement. Furthermore, administration of the dopamine receptor agonist apomorphine to lesioned animals suppressed beta-frequency oscillations, and increased coherent activity at higher frequencies in the cortex and STN, before producing the rotational behaviour indicative of successful lesion. Taken together, these results support a crucial role for dopamine in the modulation of population activity in cortico-basal ganglia circuits, whereby dopaminergic mechanisms effectively filter out synchronized, rhythmic activity at beta-frequencies at the systems level, and shift temporal couplings in these circuits to higher frequencies. These changes may be important in regulating movement. PMID- 15813952 TI - Involvement of rostral prefrontal cortex in selection between stimulus-oriented and stimulus-independent thought. AB - We used functional magnetic resonance imaging to investigate brain activity while healthy subjects performed three different tasks, each of which alternated between: (i) phases relying on stimulus-oriented thought (i.e. cognitive processes provoked by incoming sensory information); and (ii) phases relying on stimulus-independent thought (i.e. cognitive processes that were not related to any information in the immediate sensory environment). Within each task, the two phases were matched as closely as possible. In all three tasks, lateral rostral prefrontal cortex was transiently activated by a switch between stimulus-oriented and stimulus-independent thought (regardless of the direction of the switch). Medial rostral prefrontal cortex consistently exhibited sustained activity for stimulus-oriented vs. stimulus-independent thought. These results suggest the involvement of rostral prefrontal cortex in selection between stimulus-oriented and stimulus-independent cognitive processes. PMID- 15813953 TI - NR2 to NR3B subunit switchover of NMDA receptors in early postnatal motoneurons. AB - The NR3B NMDA receptor subunit is selective to somatic motoneurons in the adult nervous system. Here we report its developmental expression in the mouse brain and spinal cord by in situ hybridization. NR3B mRNA was detected in few neural regions during embryonic and neonatal periods. It first appeared in motoneurons at postnatal day (P)10-P14, and attained the maximal level at P21 and adult stage. This developmental profile was reciprocal with that of NR2 subunits, of which NR2A mRNA was most predominant in embryonic and neonatal motoneurons and downregulated by P14. Interestingly, mRNA of the NR1 subunit, which is required for functional NMDA receptors, displayed a 'V'-shaped change, decreasing with the early postnatal decline of NR2 mRNAs and increasing with the subsequent appearance of NR3B mRNA. Therefore, the major regulatory subunit of NMDA receptors is likely to switch from NR2 to NR3B in somatic motoneurons during the early postnatal period. PMID- 15813954 TI - The effect of aging on the subcellular distribution of estrogen receptor-alpha in the cholinergic neurons of transgenic and wild-type mice. AB - The degeneration of the basal forebrain cholinergic system plays an important role in cognitive deterioration in aging and Alzheimer's disease. Brain cholinergic neurons and their projections are affected by changes in the circulating levels of estrogens, which exert their effects mainly through the estrogen receptors. In this study, we investigated the effect of aging, estrogen status and transgenic genotype on the number of cholinergic neurons and the estrogen receptor alpha (ERalpha) content in the medial septum-vertical limb of the diagonal band of Broca. We used 6- and 12-month-old female double transgenic mice carrying mutated human amyloid precursor protein (APPswe) and presenilin-1 (PS1-A246E), and their nontransgenic littermate controls, which had been sham operated or ovariectomized at the age of 3 months. Brain sections were double immunostained for choline acetyltransferase (ChAT) and ERalpha and used for stereological cell counting. We found that the number of ChAT-immunoreactive (ir) neurons containing nuclear ERalpha-ir was significantly lower in 12- than in 6 month-old mice. However, the age of the mice, the transgenic genotype or ovariectomy had no effect on the total number of ChAT-ir neurons, or on the number and percentage of all ChAT-ir neurons that contained ERalpha. These results indicate that aging is associated with translocation of ERalphas from the nucleus to the cytoplasm. We propose that this phenomenon is linked to those age related processes known to be involved in inhibiting ERalpha binding to nuclei. PMID- 15813955 TI - Soluble guanylyl cyclase appears in a specific subset of periglomerular cells in the olfactory bulb. AB - In the brain, nitric oxide acts as an atypical messenger in cellular nonsynaptic transmission. In the olfactory bulb, this gas is produced at the level of the olfactory glomeruli by a subpopulation of periglomerular cells that participates in the first synaptic relay of the olfactory information between the olfactory nerve and the dendritic tufts of principal cells. It has been proposed that nitric oxide modulates intraglomerular synaptic integration of sensory inputs, but its specific role in the glomerular circuitry remains to be understood. In this article, we demonstrate that, in the glomerular circuits, a specific subset of periglomerular cells, most of them expressing the calcium binding protein calbindin D-28 k, expresses the beta1 subunit of the soluble guanylyl cyclase. These cells could be the targets for the action of nitric oxide at the glomerular level via activation of soluble guanylyl cyclase and production of cGMP. PMID- 15813956 TI - Differential expression of KCC2 accounts for the differential GABA responses between relay and intrinsic neurons in the early postnatal rat olfactory bulb. AB - The rat olfactory bulb is anatomically immature at birth, and considerable neurogenesis and synaptogenesis are known to take place postnatally. In addition, significant physiological changes have also been reported in this period. For example, granule cell-mediated inhibition following electrical stimulations to the lateral olfactory tract is robust during the first postnatal week, and then decreases abruptly after the second week. However, the mechanism underlying this enhanced inhibition remains to be elucidated. To know the cause of this phenomenon, we investigated the expression patterns of cation-Cl(-) co transporters (KCC1, KCC2 and NKCC1) mRNAs, which are responsible for the regulation of [Cl(-)](i). In addition, responses to gamma-aminobutyric acid (GABA) were measured by gramicidin-perforated patch-clamp recordings and Ca(2+) imaging using fura-2. We found that in the early postnatal period, mitral cells expressing KCC2 mRNA were inhibited by GABA, while granule cells lacking KCC2 mRNA expression were depolarized or excited by GABA. These results indicate that transient GABA-mediated excitation on granule cells might be the main cause of the enhanced inhibition on mitral cells, and suggest that these differential GABA responses between relay and intrinsic neurons play pivotal roles in the early postnatal rat olfactory bulb. PMID- 15813958 TI - A transcription factor for cold sensation! AB - The ability to feel hot and cold is critical for animals and human beings to survive in the natural environment. Unlike other sensations, the physiology of cold sensation is mostly unknown. In the present study, we use genetically modified mice that do not express nerve growth factor-inducible B (NGFIB) to investigate the possible role of NGFIB in cold sensation. We found that genetic deletion of NGFIB selectively affected behavioral responses to cold stimuli while behavioral responses to noxious heat or mechanical stimuli were normal. Furthermore, behavioral responses remained reduced or blocked in NGFIB knockout mice even after repetitive application of cold stimuli. Our results provide strong evidence that the first transcription factor NGFIB determines the ability of animals to respond to cold stimulation. PMID- 15813957 TI - Unaltered TNF-alpha production by macrophages and monocytes in diet-induced obesity in the rat. AB - BACKGROUND: Recent findings have established an association between obesity and immune dysfunction. However, most of the studies investigating the effects of obesity on immune function have been carried out in genetically obese rodent models. Since human obesity is mostly due to intake of a high fat diet and decreased energy expenditure, we asked whether immunological defects also occur in diet-induced obesity. Specifically, we focused on the function of monocytes and macrophages, as these cells are thought to be involved in the low-grade inflammation present in obesity. METHODS: Male Sprague-Dawley rats were fed a high-fat or a standard chow diet for either 2 or 10 weeks. At the end of the intervention period animals were anaesthetised, blood collected for determination of plasma mediator concentrations and lipopolysaccharide (LPS) stimulated production of TNF-alpha by monocytes. LPS stimulated production of TNF-alpha in alveolar macrophages was also determined. RESULTS: High-fat feeding for either 2 or 10 weeks resulted in significant increases in fat mass and serum leptin. Although increased serum leptin has previously been linked to modulation of innate immunity, we found no significant difference in the LPS stimulated production of TNF-alpha by either blood monocytes or alveolar macrophages between the dietary groups. Furthermore, we failed to find a significant increase in circulating TNF-alpha concentrations in obese animals, as reported for genetically obese animals. CONCLUSION: Our data suggest that defects in innate immune function observed in genetically obese animals are not mimicked by dietary obesity, and may more likely reflect the gross abnormality in leptin function of these models. Further work is required delineate the effects of dietary obesity on inflammatory state and immune function. PMID- 15813959 TI - Contribution of CaMKIV to injury and fear-induced ultrasonic vocalizations in adult mice. AB - Calcium-calmodulin dependent protein kinase IV (CaMKIV) is a protein kinase that activates the transcription factor CREB. Our previous work demonstrated that mice lacking CaMKIV had a defect in fear memory while behavioral responses to noxious stimuli were unchanged. Here, we measured ultrasonic vocalizations (USVs) before and after fear conditioning and in response to a noxious injection of capsaicin to measure behavioral responses to emotional stimuli. Consistent with previous findings, behavioral nociceptive responses to capsaicin were undistinguishable between wild-type and CaMKIV-/- mice. Wild-type animals showed a selective increase in 50 kHz USVs in response to capsaicin while such an increase was absent in CaMKIV-/- mice. The foot shock given during fear conditioning caused an increase in 30 kHz USVs in both wild-type and CaMKIV-/- mice. When returned to the context one hour later, USVs from the wild-type were significantly decreased. Additionally, the onset of a tone, which had previously been paired with the foot shock, caused a significant decrease in USVs during auditory conditioning. CaMKIV /- mice showed significantly less reduction in USVs when placed in the same context three days after receiving the shock, consistent with the decrease in freezing reported previously. Our results provide a new approach for investigating the molecular mechanism for emotional vocalization in mice and suggest that CaMKIV dependent signaling pathways play an important role in the emotional response to pain and fear. PMID- 15813960 TI - New players tip the scales in the balance between excitatory and inhibitory synapses. AB - Synaptogenesis is a highly controlled process, involving a vast array of players which include cell adhesion molecules, scaffolding and signaling proteins, neurotransmitter receptors and proteins associated with the synaptic vesicle machinery. These molecules cooperate in an intricate manner on both the pre- and postsynaptic sides to orchestrate the precise assembly of neuronal contacts. This is an amazing feat considering that a single neuron receives tens of thousands of synaptic inputs but virtually no mismatch between pre- and postsynaptic components occur in vivo. One crucial aspect of synapse formation is whether a nascent synapse will develop into an excitatory or inhibitory contact. The tight control of a balance between the types of synapses formed regulates the overall neuronal excitability, and is thus critical for normal brain function and plasticity. However, little is known about how this balance is achieved. This review discusses recent findings which provide clues to how neurons may control excitatory and inhibitory synapse formation, with focus on the involvement of the neuroligin family and PSD-95 in this process. PMID- 15813961 TI - Effects of particulate matter on the pulmonary and vascular system: time course in spontaneously hypertensive rats. AB - BACKGROUND: This study was performed within the scope of two multi-center European Commission-funded projects (HEPMEAP and PAMCHAR) concerning source composition-toxicity relationship for particulate matter (PM) sampled in Europe. The present study aimed to optimize the design for PM in vivo toxicity screening studies in terms of dose and time between a single exposure and the determination of the biological responses in a rat model mimicking human disease resulting in susceptibility to ambient PM. Dust in thoracic PM size-range (aerodynamic diameter <10 mum) was sampled nearby a road tunnel (RTD) using a high volume cascade impactor. Spontaneously hypertensive rats were exposed to urban dust collected in Ottawa, Canada (EHC-93 10 mg/kg of body weight; reference PM) or different RTD doses (0.3, 1, 3, 10 mg/kg of body weight) by intratracheal instillation. Necropsy was performed at 4, 24, or 48 hr after exposure. RESULTS: The neutrophil numbers in bronchoalveolar lavage fluid increased tremendously after exposure to the highest RTD doses or EHC-93. Furthermore, PM exposure slightly affected blood coagulation since there was a small but significant increase in the plasma fibrinogen levels (factor 1.2). Pulmonary inflammation and oxidative stress as well as changes in blood coagulation factors and circulating blood cell populations were observed within the range of 3 to 10 mg PM/kg of body weight without significant pulmonary injury. CONCLUSION: The optimal dose for determining the toxicity ranking of ambient derived PM samples in spontaneously hypertensive rats is suggested to be between 3 and 10 mg PM/kg of body weight under the conditions used in the present study. At a lower dose only some inflammatory effects were detected, which will probably be too few to be able to discriminate between PM samples while a completely different response pattern was observed with the highest dose. In addition to the dose, a 24-hr interval from exposure to sacrifice seemed appropriate to assess the relative toxic potency of PM since the majority of the health effects were observed one day after PM exposure compared to the other times examined. The aforementioned considerations provide a good basis for conducting PM toxicity screening studies in spontaneously hypertensive rats. PMID- 15813962 TI - Impact of tire debris on in vitro and in vivo systems. AB - BACKGROUND: It is estimated that over 80% of respirable particulate matter (PM10) in cities comes from road transport and that tire and brake wear are responsible for the 3-7% emission of it. Data on the indicators of environmental impact of tire debris (TD), originated from the tire abrasion on roads, are extremely scarce, even though TD contains chemicals (zinc and organic compounds) which can be released in the environment. METHODS: TD particle morphology was analysed with SEM, TEM and FIB instruments. TD eluates and TD organic extracts were tested at dilution series on human cell lines and Xenopus laevis embryos. 50 and 100 g/L TD were used for the eluates obtained after 24 h at pH 3 and the quantity of zinc present was measured with a ICP-AES. Eluates diluted to 1%, 10%, 50% in culture media and undiluted were used on X. laevis embryos in the FETAX test. HepG2 cells were exposed for 24 h to 0.05 - 50 mug/ml of zinc salt while A549 cells were exposed for 24, 48 and 72 h to 10, 50, 60, or 75 mug/ml of TD extract. X. laevis embryos were exposed to 50, 80, 100, or 120 mug/ml TD extract. RESULTS: The solution of undiluted 50 g/L TD produced 80.2% mortality (p < 0.01) in X. laevis embryos and this toxic effect was three times greater than that produced by 100 g/L TD. Zn accumulation in HepG2 cells was evident after 4 h exposure. A549 cells exposed to TD organic extract for 72 h presented a modified morphology, a decrease in cell proliferation and an increase in DNA damage as shown by comet assay. The dose 80 mug/ml of TD extract produced 14.6% mortality in X. laevis embryos and 15.9% mortality at 120 mug/ml. Treatment with 80, 100, or 120 mug/ml TD organic extract increased from 14.8% to 37.8% malformed larvae percentages compared to 5.6% in the control. CONCLUSION: Since the amount of Zn leached from TD is related to pH, aggregation of particles and elution process, the quantity of TD present in the environment has to be taken into account. Moreover the atmospheric conditions, which may deeply influence the particle properties, have to be considered. The TD organic fraction was toxic for cells and organisms. Thus, because of its chemical components, TD may have a potential environmental impact and has to be further investigated. PMID- 15813963 TI - Selective activation of primary afferent fibers evaluated by sine-wave electrical stimulation. AB - Transcutaneous sine-wave stimuli at frequencies of 2000, 250 and 5 Hz (Neurometer) are thought to selectively activate Abeta, Adelta and C afferent fibers, respectively. However, there are few reports to test the selectivity of these stimuli at the cellular level. In the present study, we analyzed action potentials (APs) generated by sine-wave stimuli applied to the dorsal root in acutely isolated rat dorsal root ganglion (DRG) preparations using intracellular recordings. We also measured excitatory synaptic responses evoked by transcutaneous stimuli in substantia gelatinosa (SG) neurons of the spinal dorsal horn, which receive inputs predominantly from C and Adelta fibers, using in vivo patch-clamp recordings. In behavioral studies, escape or vocalization behavior of rats was observed with both 250 and 5 Hz stimuli at intensity of approximately 0.8 mA (T5/ T250), whereas with 2000 Hz stimulation, much higher intensity (2.14 mA, T2000) was required. In DRG neurons, APs were generated at T5/T250 by 2000 Hz stimulation in Abeta, by 250 Hz stimulation both in Abeta and Adelta, and by 5 Hz stimulation in all three classes of DRG neurons. However, the AP frequencies elicited in Abeta and Adelta by 5 Hz stimulation were much less than those reported previously in physiological condition. With in vivo experiments large amplitude of EPSCs in SG neurons were elicited by 250 and 5 Hz stimuli at T5/ T250. These results suggest that 2000 Hz stimulation excites selectively Abeta fibers and 5 Hz stimulation activates noxious transmission mediated mainly through C fibers. Although 250 Hz stimulation activates both Adelta and Abeta fibers, tactile sensation would not be perceived when painful sensation is produced at the same time. Therefore, 250 Hz was effective stimulus frequency for activation of Adelta fibers initiating noxious sensation. Thus, the transcutaneous sine-wave stimulation can be applied to evaluate functional changes of sensory transmission by comparing thresholds with the three stimulus frequencies. PMID- 15813964 TI - A pain in the ACC. AB - An emerging theme in systems neurobiology is that even simple forms of memory depend on activity in a broad network of cortical and subcortical brain regions. One key challenge is to understand how different components of these complex networks contribute to memory. In a new study in Molecular Pain, Tang and colleagues use a novel set of approaches to characterize the role of the anterior cingulate cortex (ACC) in the formation of Pavlovian fear memories. PMID- 15813965 TI - Trapping of normal EB1 ligands in aggresomes formed by an EB1 deletion mutant. AB - BACKGROUND: EB1 is a microtubule tip-associated protein that interacts with the APC tumour suppressor protein and the p150glued subunit of dynactin. We previously reported that an EB1 deletion mutant that retains both of these interactions but does not directly associate with microtubules (EB1-DeltaN2-GFP) spontaneously formed perinuclear aggregates when expressed in COS-7 cells. RESULTS: In the present study live imaging indicated that EB1-DeltaN2-GFP aggregates underwent dynamic microtubule-dependent changes in morphology and appeared to be internally cohesive. EB1-DeltaN2-GFP aggregates were phase-dense structures that displayed microtubule-dependent accumulation around the centrosome, were immunoreactive for both the 20s subunit of the proteasome and ubiquitin, and induced the collapse of the vimentin cytoskeleton. Fractionation studies revealed that a proportion of EB1-DeltaN2-GFP was detergent-insoluble and ubiquitylated, indicating that EB1-DeltaN2-GFP aggregates are aggresomes. Immunostaining also revealed that APC and p150glued were present in EB1-DeltaN2 GFP aggregates, whereas EB3 was not. Furthermore, evidence for p150glued degradation was found in the insoluble fraction of EB1-DeltaN2-GFP transfected cultures. CONCLUSION: Our data indicate that aggresomes can be internally cohesive and may not represent a simple "aggregate of aggregates" assembled around the centrosome. Our observations also indicate that a partially misfolded protein may retain the ability to interact with its normal physiological ligands, leading to their co-assembly into aggresomes. This supports the idea that the trapping and degradation of co-aggregated proteins might contribute to human pathologies characterised by aggresome formation. PMID- 15813966 TI - Focusing in bioproduction science. AB - As in other Biotechnological fields, the microbial production of recombinant proteins and other biomolecules can be approached from multiple angles through the help of diverse technologies of increasing complexity. To better reach all the specialized niches in bioproduction, Microbial Cell Factories is now inviting authors to prepare concise Reviews (eventually miniReviews), covering relevant areas that deserve specific and highly focused attention. By the publication of such contributions, the journal will promote the revision of new insights around the Cell Factory concept in a highly comprehensive way, in molecular, cellular and environmental contexts. PMID- 15813967 TI - Androgen receptor signaling is required for androgen-sensitive human prostate cancer cell proliferation and survival. AB - BACKGROUND: Androgens and androgen receptors (AR) regulate normal prostate development and growth. They also are involved in pathological development of prostatic diseases, including benign prostatic hyperplasia (BPH) and prostate cancer (PCa). Antiandrogen therapy for PCa, in conjunction with chemical or surgical castration, offers initial positive responses and leads to massive prostate cell death. However, cancer cells later appear as androgen-independent PCa. To investigate the role of AR in prostate cell proliferation and survival, we introduced a vector-based small interfering RNA (siRNA). This siRNA targeted 5'-untranslated region of AR mRNA for extended suppression of AR expression in androgen-sensitive human prostate LNCaP cells. RESULTS: The siRNA design successfully suppressed endogenous AR expression, as revealed by western blotting and immunofluorescence staining in LNCaP cells. LNCaP cells did not proliferate in the absence of AR and underwent apoptosis, based on elevated phospho-Histone H2B expression and higher number of apoptotic body as compared to control cells. CONCLUSION: We demonstrated that AR is vital for prostate cell proliferation and survival in this androgen-sensitive prostate cell line. These results further strengthen the hypothesis that AR can be a therapeutic target for treating androgen-sensitive stages of PCa. Unlike antiandorgens, however, siRNA targeting AR provides a direct inactivation of AR function through the suppression of AR protein expression. PMID- 15813969 TI - First Dominique Dormont International Conference on "Host-pathogen interactions in chronic infections - viral and host determinants of HCV, HCMV, and HIV infections". AB - The first Dominique Dormont International Conference on "Viral and host determinantsof HCV, HCMV, and HIV infections "was held in Paris, Val-de-Grace, on December 3-4, 2004. The following is a summary of the scientific sessions of this meeting (http://www.congres-evenement.fr/ddormont). PMID- 15813968 TI - HDBStat!: a platform-independent software suite for statistical analysis of high dimensional biology data. AB - BACKGROUND: Many efforts in microarray data analysis are focused on providing tools and methods for the qualitative analysis of microarray data. HDBStat! (High Dimensional Biology-Statistics) is a software package designed for analysis of high dimensional biology data such as microarray data. It was initially developed for the analysis of microarray gene expression data, but it can also be used for some applications in proteomics and other aspects of genomics. HDBStat! provides statisticians and biologists a flexible and easy-to-use interface to analyze complex microarray data using a variety of methods for data preprocessing, quality control analysis and hypothesis testing. RESULTS: Results generated from data preprocessing methods, quality control analysis and hypothesis testing methods are output in the form of Excel CSV tables, graphs and an Html report summarizing data analysis. CONCLUSION: HDBStat! is a platform-independent software that is freely available to academic institutions and non-profit organizations. It can be downloaded from our website http://www.soph.uab.edu/ssg_content.asp?id=1164. PMID- 15813970 TI - Anti-inflammatory therapy by ibudilast, a phosphodiesterase inhibitor, in demyelination of twitcher, a genetic demyelination model. AB - BACKGROUND: Twitcher mouse (twi/twi) is an authentic murine model of Krabbe's disease. Accumulation of psychosine, resulting in apoptosis of oligodendrocytes and subsequent demyelination, is a cardinal event to the pathogenesis of this disease. Moreover, recruitment of inflammatory cells plays a significant role in the pathological process in the twi/twi central and peripheral nervous systems. In this study, we investigated the 1) the relationship between tumor necrosis factor-alpha (TNFalpha), pro-inflammatory cytokine, and the progression of this disease and 2) effect of the anti-inflammatory therapy by ibudilast, a phosphodiesterase inhibitor. METHODS: We quantified the expression level of TNFalpha and TNF-receptor mRNA in twi/twi using semi-quantitative RT-PCR. The relationship between TNFalpha expression, apoptosis of oligodendrocytes and demyelination was studied with immunohistochemistry and TUNEL method. We then treated twi/twi with a daily intraperitoneal injection of ibudilast (10 mg/kg), which suppress TNFalpha production in the brain. RESULTS: We found that TNFalpha immunoreactive microglia/macrophages appeared in the twi/twi brain and that the mRNA levels of TNFalpha and TNF-receptor 1 was increased with the progression of demyelination. The distribution profile of TNFalpha-immunoreactive microglia/macrophages overlapped that of TUNEL-positive oligodendrocytes in the twi/twi brain. When twi/twi was treated with ibudilast from PND30, the number of oligodendrocytes undergoing apoptosis was markedly reduced and demyelination was milder. Obvious improvement of clinical symptom was noted in two of five. The failure of constant clinical improvement by ibudilast may result from hepatotoxicity and/or the inhibition of proliferation of NG2-positive oligodendrocyte precursors. CONCLUSION: We conclude that anti-inflammatory therapy by a phosphodiesterase inhibitor can be considered as a novel alternative therapy for Krabbe's disease. PMID- 15813972 TI - Automated analysis of digital fundus autofluorescence images of geographic atrophy in advanced age-related macular degeneration using confocal scanning laser ophthalmoscopy (cSLO). AB - BACKGROUND: Fundus autofluorescence (AF) imaging using confocal scanning laser ophthalmoscopy (cSLO) provides an accurate delineation of areas of geographic atrophy (GA). Automated computer-assisted methods for detecting and removing interfering vessels are needed to support the GA quantification process in longitudinal studies and in reading centres. METHODS: A test tool was implemented that uses region-growing techniques to segment GA areas. An algorithm for illuminating shadows can be used to process low-quality images. Agreement between observers and between three different methods was evaluated by two independent readers in a pilot study. Agreement and objectivity were assessed using the Bland Altman approach. RESULTS: The new method (C) identifies vascular structures that interfere with the delineation of GA. Results are comparable to those of two commonly used procedures (A, B), with a mean difference between C and A of -0.67 mm2 (95% CI [-0.99, -0.36]), between B and A of -0.81 mm2, (95% CI [-1.08, 0.53]), and between C and B of 0.15 mm2 (95% CI [-0.12, 0.41]). Objectivity of a method is quantified by the mean difference between observers: A 0.30 mm2 (95% CI [0.02, 0.57]), B -0.11 mm2 (95% CI [-0.28, 0.10]), and C 0.12 mm2 (95% CI [0.02, 0.22]). CONCLUSION: The novel procedure is comparable with regard to objectivity and inter-reader agreement to established methods of quantifying GA. It considerably speeds up the lengthy measurement process in AF with well defined GA zones. PMID- 15813971 TI - The Blood Pressure "Uncertainty Range" - a pragmatic approach to overcome current diagnostic uncertainties (II). AB - A tremendous amount of scientific evidence regarding the physiology and physiopathology of high blood pressure combined with a sophisticated therapeutic arsenal is at the disposal of the medical community to counteract the overall public health burden of hypertension. Ample evidence has also been gathered from a multitude of large-scale randomized trials indicating the beneficial effects of current treatment strategies in terms of reduced hypertension-related morbidity and mortality.In spite of these impressive advances and, deeply disappointingly from a public health perspective, the real picture of hypertension management is overshadowed by widespread diagnostic inaccuracies (underdiagnosis, overdiagnosis) as well as by treatment failures generated by undertreatment, overtreatment, and misuse of medications.The scientific, medical and patient communities as well as decision-makers worldwide are striving for greatest possible health gains from available resources.A seemingly well-crystallised reasoning is that comprehensive strategic approaches must not only target hypertension as a pathological entity, but rather, take into account the wider environment in which hypertension is a major risk factor for cardiovascular disease carrying a great deal of our inheritance, and its interplay in the constellation of other, well-known, modifiable risk factors, i.e., attention is to be switched from one's "blood pressure level" to one's absolute cardiovascular risk and its determinants. Likewise, a risk/benefit assessment in each individual case is required in order to achieve best possible results.Nevertheless, it is of paramount importance to insure generalizability of ABPM use in clinical practice with the aim of improving the accuracy of a first diagnosis for both individual treatment and clinical research purposes. Widespread adoption of the method requires quick adjustment of current guidelines, development of appropriate technology infrastructure and training of staff (i.e., education, decision support, and information systems for practitioners and patients). Progress can be achieved in a few years, or in the next 25 years. PMID- 15813973 TI - Sildenafil citrate increases myocardial cGMP content in rat heart, decreases its hypertrophic response to isoproterenol and decreases myocardial leak of creatine kinase and troponin T. AB - BACKGROUND: Cardiac hypertrophy is a major risk factor for morbidity and mortality in a number of cardiovascular diseases. Consequently, the signaling pathways that inhibit cardiac hypertrophy are currently receiving much interest. Among them, nitric oxide (NO), signaling via cGMP and cGMP-dependent protein kinase I, has been recognized as a negative regulator of cardiac hypertrophy. The present study investigated the in-vivo effect of sildenafil as a phosphodiestrase 5A (PDE-5A) inhibitor on the hypertrophic response of rat heart to isoproterenol and the relation of this effect to the level of myocardial cGMP and integrity of the constitutive nitric oxide synthase (cNOS) activity. RESULTS: The results showed that daily intraperitoneal administration of sildenafil per se for 10 days was without noticeable adverse effects on survival or myocardium. Conversely, daily subcutaneous administration of isoproterenol for 10 days caused significant myocardial hypertrophy, cell injury and decline in survival. When sildenafil was injected daily, one hour before isoproterenol, survival was significantly improved and the myocardium didn't show significant hypertrophy or cell injury. Interestingly, sildenafil was accompanied by significant rise in myocardial cGMP level, a parameter which was found in the present study to possess a significant negative correlation with cardiac hypertrophy and leak of cardiac troponin T into serum. At the same time, cGMP was found to possess a positive correlation with myocardial creatine kinase activity that reflects the efficiency of the energy utilization processes in the myocardium. However, in rats given Nomega-nitro-L arginine (L-NNA) as a competitive inhibitor of cNOS, sildenafil failed to show any favorable effect on survival or the myocardial injury parameters used to assess isoproterenol-induced injury. CONCLUSION: The present study suggests that increased cardiac cGMP level by sildenafil have a cardioprotective effect probably through acting as a post-receptor negative regulator of cardiac sympathetic responsiveness. Integrity of NOS function was an essential prerequisite for sildenafil's mediated cardioprotection encountered in the present study. PMID- 15813974 TI - Vertical distribution of zooplankton: density dependence and evidence for an ideal free distribution with costs. AB - BACKGROUND: In lakes with a deep-water algal maximum, herbivorous zooplankton are faced with a trade-off between high temperature but low food availability in the surface layers and low temperature but sufficient food in deep layers. It has been suggested that zooplankton (Daphnia) faced with this trade-off distribute vertically according to an "Ideal Free Distribution (IFD) with Costs". An experiment has been designed to test the density (competition) dependence of the vertical distribution as this is a basic assumption of IFD theory. RESULTS: Experiments were performed in large, indoor mesocosms (Plankton Towers) with a temperature gradient of 10 degrees C and a deep-water algal maximum established below the thermocline. As expected, Daphnia aggregated at the interface between the two different habitats when their density was low. The distribution spread asymmetrically towards the algal maximum when the density increased until 80 % of the population dwelled in the cool, food-rich layers at high densities. Small individuals stayed higher in the water column than large ones, which conformed with the model for unequal competitors. CONCLUSION: The Daphnia distribution mimics the predictions of an IFD with costs model. This concept is useful for the analysis of zooplankton distributions under a large suite of environmental conditions shaping habitat suitability. Fish predation causing diel vertical migrations can be incorporated as additional costs. This is important as the vertical location of grazing zooplankton in a lake affects phytoplankton production and species composition, i.e. ecosystem function. PMID- 15813976 TI - Workflows in bioinformatics: meta-analysis and prototype implementation of a workflow generator. AB - BACKGROUND: Computational methods for problem solving need to interleave information access and algorithm execution in a problem-specific workflow. The structures of these workflows are defined by a scaffold of syntactic, semantic and algebraic objects capable of representing them. Despite the proliferation of GUIs (Graphic User Interfaces) in bioinformatics, only some of them provide workflow capabilities; surprisingly, no meta-analysis of workflow operators and components in bioinformatics has been reported. RESULTS: We present a set of syntactic components and algebraic operators capable of representing analytical workflows in bioinformatics. Iteration, recursion, the use of conditional statements, and management of suspend/resume tasks have traditionally been implemented on an ad hoc basis and hard-coded; by having these operators properly defined it is possible to use and parameterize them as generic re-usable components. To illustrate how these operations can be orchestrated, we present GPIPE, a prototype graphic pipeline generator for PISE that allows the definition of a pipeline, parameterization of its component methods, and storage of metadata in XML formats. This implementation goes beyond the macro capacities currently in PISE. As the entire analysis protocol is defined in XML, a complete bioinformatic experiment (linked sets of methods, parameters and results) can be reproduced or shared among users. AVAILABILITY: http://if web1.imb.uq.edu.au/Pise/5.a/gpipe.html (interactive), ftp://ftp.pasteur.fr/pub/GenSoft/unix/misc/Pise/ (download). CONCLUSION: From our meta-analysis we have identified syntactic structures and algebraic operators common to many workflows in bioinformatics. The workflow components and algebraic operators can be assimilated into re-usable software components. GPIPE, a prototype implementation of this framework, provides a GUI builder to facilitate the generation of workflows and integration of heterogeneous analytical tools. PMID- 15813975 TI - Beyond the Evidence of the New Hypertension Guidelines. Blood pressure measurement - is it good enough for accurate diagnosis of hypertension? Time might be in, for a paradigm shift (I). AB - Despite widespread availability of a large body of evidence in the area of hypertension, the translation of that evidence into viable recommendations aimed at improving the quality of health care is very difficult, sometimes to the point of questionable acceptability and overall credibility of the guidelines advocating those recommendations.The scientific community world-wide and especially professionals interested in the topic of hypertension are witnessing currently an unprecedented debate over the issue of appropriateness of using different drugs/drug classes for the treatment of hypertension. An endless supply of recent and less recent "drug-news", some in support of, others against the current guidelines, justifying the use of selected types of drug treatment or criticising other, are coming out in the scientific literature on an almost weekly basis. The latest of such debate (at the time of writing this paper) pertains the safety profile of ARBs vs ACE inhibitors.To great extent, the factual situation has been fuelled by the new hypertension guidelines (different for USA, Europe, New Zeeland and UK) through, apparently small inconsistencies and conflicting messages, that might have generated substantial and perpetuating confusion among both prescribing physicians and their patients, regardless of their country of origin.The overwhelming message conveyed by most guidelines and opinion leaders is the widespread use of diuretics as first-line agents in all patients with blood pressure above a certain cut-off level and the increasingly aggressive approach towards diagnosis and treatment of hypertension. This, apparently well-justified, logical and easily comprehensible message is unfortunately miss-obeyed by most physicians, on both parts of the Atlantic.Amazingly, the message assumes a universal simplicity of both diagnosis and treatment of hypertension, while ignoring several hypertension-specific variables, commonly known to have high level of complexity, such as:- accuracy of recorded blood pressure and the great inter-observer variability,- diversity in the competency and training of diagnosing physician,- individual patient/disease profile with highly subjective preferences,- difficulty in reaching consensus among opinion leaders,- pharmaceutical industry's influence, and, nonetheless,- the large variability in the efficacy and safety of the antihypertensive drugs.The present 2-series article attempts to identify and review possible causes that might have, at least in part, generated the current healthcare anachronism (I); to highlight the current trend to account for the uncertainties related to the fixed blood pressure cut-off point and the possible solutions to improve accuracy of diagnosis and treatment of hypertension (II). PMID- 15813977 TI - Risk factors for fatal candidemia caused by Candida albicans and non-albicans Candida species. AB - BACKGROUND: Invasive fungal infections, such as candidemia, caused by Candida species have been increasing. Candidemia is not only associated with a high mortality (30% to 40%) but also extends the length of hospital stay and increases the costs of medical care. Sepsis caused by Candida species is clinically indistinguishable from bacterial infections. Although, the clinical presentations of the patients with candidemia caused by Candida albicans and non-albicans Candida species (NAC) are indistinguishable, the susceptibilities to antifungal agents of these species are different. In this study, we attempted to identify the risk factors for candidemia caused by C. albicans and NAC in the hope that this may guide initial empiric therapy. METHODS: A retrospective chart review was conducted during 1996 to 1999 at the Veterans General Hospital-Taipei. RESULTS: There were 130 fatal cases of candidemia, including 68 patients with C. albicans and 62 with NAC. Candidemia was the most likely cause of death in 55 of the 130 patients (42.3 %). There was no significant difference in the distribution of Candida species between those died of candidemia and those died of underlying conditions. Patients who had one of the following conditions were more likely to have C. albicans, age > or = 65 years, immunosuppression accounted to prior use of steroids, leukocytosis, in the intensive care unit (ICU), and intravascular and urinary catheters. Patients who had undergone cancer chemotherapy often appeared less critically ill and were more likely to have NAC. CONCLUSION: Clinical and epidemiological differences in the risk factors between candidemia caused by C. albicans and NAC may provide helpful clues to initiate empiric therapy for patients infected with C. albicans versus NAC. PMID- 15813978 TI - The role of mast cells and fibre type in ischaemia reperfusion injury of murine skeletal muscles. AB - BACKGROUND: Ischaemia reperfusion (IR) injury of skeletal muscle, is a significant cause of morbidity following trauma and surgical procedures, in which muscle fibre types exhibit different susceptibilities. The relative degree of mast cell mediated injury, within different muscle types, is not known. METHODS: In this study we compared susceptibility of the fast-twitch, extensor digitorum longus (EDL), mixed fast/slow-twitch gastrocnemius and the predominately slow twitch soleus, muscles to ischemia reperfusion (IR) injury in four groups of mice that harbour different mast cell densities; C57/DBA mast cell depleted (Wf/Wf), their heterozygous (Wf/+) and normal littermates (+/+) and control C57BL/6 mice. We determined whether susceptibility to IR injury is associated with mast cell content and/or fibre type and/or mouse strain. In experimental groups, the hind limbs of mice were subjected to 70 minutes warm tourniquet ischemia, followed by 24 h reperfusion, and the muscle viability was assessed on fresh whole-mount slices by the nitroblue tetrazolium (NBT) histochemical assay. RESULTS: Viability was remarkably higher in the Wf/Wf strain irrespective of muscle type. With respect to muscle type, the predominately slow-twitch soleus muscle was significantly more resistant to IR injury than gastrocnemius and the EDL muscles in all groups. Mast cell density was inversely correlated to muscle viability in all types of muscle. CONCLUSION: These results show that in skeletal muscle, IR injury is dependent upon both the presence of mast cells and on fibre type and suggest that a combination of preventative therapies may need to be implemented to optimally protect muscles from IR injury. PMID- 15813979 TI - The Journal of Inflammation. AB - Welcome to the Journal of Inflammation, the first open-access, peer-reviewed, online journal to focus on all aspects of the study of inflammation and inflammatory conditions. While research into inflammation has resulted in great progress in the latter half of the 20th century, the rate of progress is rapidly accelerating. Thus there is a need for a vehicle through which this very diverse research can be made readily available to the scientific community. The Journal of Inflammation, a peer reviewed journal, provides the ideal vehicle for such rapid dissemination of information. The Journal of Inflammation covers the full range of underlying cellular and molecular mechanisms involved, not only in the production of the inflammatory responses but, more importantly in clinical terms, in the healing process as well. This includes molecular, cellular, animal and clinical studies related to the study of inflammatory conditions and responses, and all related aspects of pharmacology, such as anti-inflammatory drug development, trials and therapeutic developments, etc. All articles published in the Journal of Inflammation are immediately listed in PubMed, and access to published articles is universal and free through the internet. PMID- 15813980 TI - Cancer, inflammation and the AT1 and AT2 receptors. AB - The critical role of inappropriate inflammation is becoming accepted in many diseases that affect man, including cardiovascular diseases, inflammatory and autoimmune disorders, neurodegenerative conditions, infection and cancer.This review proposes that cancer up-regulates the angiotensin II type 1 (AT1) receptor through systemic oxidative stress and hypoxia mechanisms, thereby triggering chronic inflammatory processes to remodel surrounding tissue and subdue the immune system. Based on current literature and clinical studies on angiotensin receptor inhibitors, the paper concludes that blockade of the AT1 receptor in synergy with cancer vaccines and anti-inflammatory agents should offer a therapy to regress most, if not all, solid tumours.With regard to cancer being a systemic disease, an examination of supporting evidence for a systemic role of AT1 in relationship to inflammation in disease and injury is presented as a logical progression. The evidence suggests that regulation of the mutually antagonistic angiotensin II receptors (AT1 and AT2) is an essential process in the management of inflammation and wound recovery, and that it is an imbalance in the expression of these receptors that leads to disease.In consideration of cancer induced immune suppression, it is further postulated that the inflammation associated with bacterial and viral infections, is also an evolved means of immune suppression by these pathogens and that the damage caused, although incidental, leads to the symptoms of disease and, in some cases, death.It is anticipated that manipulation of the angiotensin system with existing anti-hypertensive drugs could provide a new approach to the treatment of many of the diseases that afflict mankind. PMID- 15813981 TI - Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo. AB - BACKGROUND: RNA interference is an evolutionary conserved immune response mechanism that can be used as a tool to provide novel insights into gene function and structure. The ability to efficiently deliver small interfering RNA to modulate gene expression in vivo may provide new therapeutic approaches to currently intractable diseases. METHODS: In vitro, siRNA targeting IL-12p40 was delivered to the murine macrophage cell line (J774A.1) encapsulated in a liposome with an IL-12 inducing agent (LPS/IFN-gamma) over a number of time points. Controls included a variety of non-target specific siRNA reagents. Supernatants were analyzed for cytokine production while the cells were removed for mRNA profiling. In vivo, siRNA-targeting IL-12p40 was delivered to the murine peritoneal cavity in a therapeutic fashion, after endotoxin (LPS) challenge. Cells from the peritoneal cavity were removed by lavage and analyzed by flow cytometry. Levels of IL-12 present in lavage and in serum were also examined by ELISA. RESULTS: In this report, we show that IL-12p40 siRNA can specifically silence macrophage expression of IL-12p40 mRNA and IL-12p70 protein in vitro. We extend this finding to demonstrate that delivery of liposome encapsulated siRNA targeting IL-12p40 to the murine peritoneal cavity can modulate an inflammatory stimulus in vivo. Furthermore, specific siRNA can be used therapeutically after endotoxin challenge to reduce both the local and systemic inflammatory response. Thus, the delivery of siRNA can be used to elicit specific non-permanent inhibition of endogenous protein expression. CONCLUSION: In vitro silencing of IL 12p40 using siRNA at selected doses leads to specific knockdown of IL-12p70 protein production without inducing type I interferons. Furthermore, siRNA targeting murine IL-12p40 can be used therapeutically to counter an inflammatory response in vivo. PMID- 15813982 TI - Particle and Fibre Toxicology, a new journal to meet a real need. AB - This Editorial is to announce Particle and Fibre Toxicology, a new Open Access, peer-reviewed, online journal published by BioMed Central. The field of particle and fibre toxicology has a long and famous history stretching from Agricola and Paracelsus in the 15th and 16th century to the challenges of the 21st century nanoparticles, nanotubes and particulate matter (PM10) to name just three. Throughout this time there has been no single journal dedicated to the toxicology of particles and fibres and this is finally corrected by the launch of Particle and Fibre Toxicology. The rationale for Particle and Fibre Toxicology rests on this need for a single multi-disciplinary journal that can cover all research relevant to particle and fibre toxicology, from Hygiene studies, through particle generation and characterisation, to animal, cell and human toxicology studies, dosimetry and modelling. The editorial also deals with the philosophy and practicalities of Open Access publishing, the journal's peer-review policy and conflict-of-interest. Particle and Fibre Toxicology is aimed at bringing together multi-disciplinary research findings towards a better understanding of how particles and fibres adversely affect the lungs and the body generally. We hope that the launch of the new journal will aid in the advance of this important discipline to the greater benefit of occupational and public health and invite scientists working in this key discipline to submit their research. PMID- 15813983 TI - ROS-mediated TNF-alpha and MIP-2 gene expression in alveolar macrophages exposed to pine dust. AB - BACKGROUND: Respiratory symptoms, impaired lung function, and asthma have been reported in workers exposed to wood dust in a number of epidemiological studies. The underlying pathomechanisms, however, are not well understood. Here, we studied the effects of dust from pine (PD) and heat-treated pine (HPD) on the release of reactive oxygen species (ROS) and inflammatory mediators in rat alveolar macrophages. METHODS: Tumour necrosis factor-alpha (TNF-alpha) and macrophage inflammatory protein-2 (MIP-2) protein release, TNF-alpha and MIP-2 mRNA expression, and generation of ROS were studied as end points after treatment of rat alveolar macrophages with PD or HPD. In a separate series of experiments, the antioxidants glutathione and N-acetyl-L-cysteine were included in combination with wood dust. To determine the endogenous oxidative and antioxidant capacity of wood dusts, electron spin resonance (ESR) spectroscopy was used. RESULTS: After 4 h incubation, both PD and HPD elicited a significantly (p < 0.05) increased mRNA expression of TNF-alpha and MIP-2 as well as a concentration-dependent release of TNF-alpha and MIP-2 protein. Interestingly, PD induced a significantly higher TNF alpha and MIP-2 production than HPD. Moreover, a significantly increased ROS production was observed in alveolar macrophages exposed to both PD and HPD. In the presence of the antioxidants glutathione and N-acetyl-L-cysteine, the PD- and HPD-induced release of ROS, TNF-alpha, and MIP-2 was significantly reduced. Finally, electron spin resonance analyses demonstrated a higher endogenous antioxidant capacity of HPD compared to PD. Endotoxin was not present in either dust sample. CONCLUSION: These results indicate that pine dust is able to induce expression of TNF-alpha and MIP-2 in rat alveolar macrophages by a mechanism that is, at least in part, mediated by ROS. PMID- 15813984 TI - What does the marriage of Open Access with online publication bring? AB - Open Access online publishing is the trend of the future for unrestricted rapid and international dissemination of knowledge. Several journals are published on acquired immune deficiency syndrome (AIDS) research, but none of them appear to be Open Access. To eliminate or to abate the scourge of AIDS, it is important that the knowledge acquired through research be disseminated as soon as possible. The Open Access journal, AIDS Research and Therapy, is intended to fill this knowledge gap by online publication of basic, preclinical, and clinical research articles. PMID- 15813985 TI - Cardiovascular effects in patrol officers are associated with fine particulate matter from brake wear and engine emissions. AB - BACKGROUND: Exposure to fine particulate matter air pollutants (PM2.5) affects heart rate variability parameters, and levels of serum proteins associated with inflammation, hemostasis and thrombosis. This study investigated sources potentially responsible for cardiovascular and hematological effects in highway patrol troopers. RESULTS: Nine healthy young non-smoking male troopers working from 3 PM to midnight were studied on four consecutive days during their shift and the following night. Sources of in-vehicle PM2.5 were identified with variance-maximizing rotational principal factor analysis of PM2.5-components and associated pollutants. Two source models were calculated. Sources of in-vehicle PM2.5 identified were 1) crustal material, 2) wear of steel automotive components, 3) gasoline combustion, 4) speed-changing traffic with engine emissions and brake wear. In one model, sources 1 and 2 collapsed to a single source. Source factors scores were compared to cardiac and blood parameters measured ten and fifteen hours, respectively, after each shift. The "speed change" factor was significantly associated with mean heart cycle length (MCL, +7% per standard deviation increase in the factor score), heart rate variability (+16%), supraventricular ectopic beats (+39%), % neutrophils (+7%), % lymphocytes (-10%), red blood cell volume MCV (+1%), von Willebrand Factor (+9%), blood urea nitrogen (+7%), and protein C (-11%). The "crustal" factor (but not the "collapsed" source) was associated with MCL (+3%) and serum uric acid concentrations (+5%). Controlling for potential confounders had little influence on the effect estimates. CONCLUSION: PM2.5 originating from speed-changing traffic modulates the autonomic control of the heart rhythm, increases the frequency of premature supraventricular beats and elicits pro-inflammatory and pro-thrombotic responses in healthy young men. PMID- 15813986 TI - Lentiviral transduction of Tar Decoy and CCR5 ribozyme into CD34+ progenitor cells and derivation of HIV-1 resistant T cells and macrophages. AB - BACKGROUND: RNA based antiviral approaches against HIV-1 are among the most promising for long-term gene therapy. These include ribozymes, aptamers (decoys), and small interfering RNAs (siRNAs). Lentiviral vectors are ideal for transduction of such inhibitory RNAs into hematopoietic stem cells due to their ability to transduce non-dividing cells and their relative refractiveness to gene silencing. The objective of this study is to introduce an HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme into CD34+ hematopoietic progenitor cells via an HIV-based lentiviral vector to derive viral resistant progeny T cells and macrophages. RESULTS: High efficiency and sustained gene transfer into CD34+ cells were achieved with lentiviral vector constructs harboring either Tar decoy or Tar decoy in combination with CCR5 ribozyme. Cells transduced with these constructs differentiated normally into T-lymphocytes in vivo in thy/liv grafts of SCID-hu mice, and into macrophages in vitro in the presence of appropriate growth factors. When challenged in vitro, the differentiated T lymphocytes and macrophages showed marked resistance against HIV 1 infection. CONCLUSIONS: Viral resistant transgenic T cells and macrophages that express HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme could be obtained by lentiviral gene transduction of CD34+ progenitor cells. These results showed for the first time that expression of these anti-HIV 1 transgenes in combination do not interfere with normal thymopoiesis and thus have set the stage for their application in stem cell based gene therapy for HIV/AIDS. PMID- 15813987 TI - Molecular pain, a new era of pain research and medicine. AB - Molecular pain is a relatively new and rapidly expanding research field that represents an advanced step from conventional pain research. Molecular pain research addresses physiological and pathological pain at the cellular, subcellular and molecular levels. These studies integrate pain research with molecular biology, genomics, proteomics, modern electrophysiology and neurobiology. The field of molecular pain research has been rapidly expanding in the recent years, and has great promise for the identification of highly specific and effective targets for the treatment of intractable pain. Although several existing journals publish articles on classical pain research, none are specifically dedicated to molecular pain research. Therefore, a new journal focused on molecular pain research is needed. Molecular Pain, an Open Access, peer-reviewed, online journal, will provide a forum for molecular pain scientists to communicate their research findings in a targeted manner to others in this important and growing field. PMID- 15813988 TI - A P2X receptor-mediated nociceptive afferent pathway to lamina I of the spinal cord. AB - Of the six lamina regions in the dorsal horn of the spinal cord, lamina I is a major sensory region involved in nociceptive transmission under both physiological and pathological conditions. While P2X receptors have been shown to be involved in nociception, it remains unknown if P2X receptors are involved in nociceptive transmission to lamina I neurons. Using rat spinal cord slice preparations and patch-clamp recordings, we have demonstrated that the excitatory synaptic transmission between primary afferent fibers and lamina I neurons is significantly affected by ATP and alpha,beta-methylene-ATP. The synaptic effects of them include the increases of the frequency of both miniature excitatory postsynaptic currents (mEPSCs) and spontaneous EPSCs (sEPSCs), and decreases of evoked EPSCs (eEPSCs). These effects were blocked by pyridoxalphosphate-6 azophenyl-2', 4'-disulfonic acid (PPADS, 10 microM) and suramin (30 microM). In the neurons for which ATP and alpha,beta-methylene-ATP had effects on mEPSCs, sEPSCs and eEPSCs, capsaicin produced similar synaptic effects. Our results indicate that P2X receptors are expressed on many afferent fibers that directly synapse to lamina I neurons. Furthermore, these P2X receptor-expressing afferent fibers are capsaicin-sensitive nociceptive afferents. Thus, this study reveals a P2X receptor-mediated nociceptive afferent pathway to lamina I of the spinal cord and provides a new insight into the nociceptive functions of P2X receptors. PMID- 15813989 TI - Sensitization of TRPV1 by EP1 and IP reveals peripheral nociceptive mechanism of prostaglandins. AB - Prostaglandin E2 (PGE2) and prostaglandin I2 (PGI2) are major inflammatory mediators that play important roles in pain sensation and hyperalgesia. The role of their receptors (EP and IP, respectively) in inflammation has been well documented, although the EP receptor subtypes involved in this process and the underlying cellular mechanisms remain to be elucidated. The capsaicin receptor TRPV1 is a nonselective cation channel expressed in sensory neurons and activated by various noxious stimuli. TRPV1 has been reported to be critical for inflammatory pain mediated through PKA- and PKC-dependent pathways. PGE2 or PGI2increased or sensitized TRPV1 responses through EP1 or IP receptors, respectively predominantly in a PKC-dependent manner in both HEK293 cells expressing TRPV1 and mouse DRG neurons. In the presence of PGE2 or PGI2, the temperature threshold for TRPV1 activation was reduced below 35 degrees C, so that temperatures near body temperature are sufficient to activate TRPV1. A PKA dependent pathway was also involved in the potentiation of TRPV1 through EP4 and IP receptors upon exposure to PGE2 and PGI2, respectively. Both PGE2-induced thermal hyperalgesia and inflammatory nociceptive responses were diminished in TRPV1-deficient mice and EP1-deficient mice. IP receptor involvement was also demonstrated using TRPV1-deficient mice and IP-deficient mice. Thus, the potentiation or sensitization of TRPV1 activity through EP1 or IP activation might be one important mechanism underlying the peripheral nociceptive actions of PGE2 or PGI2. PMID- 15813990 TI - HIV-1 resistance conferred by siRNA cosuppression of CXCR4 and CCR5 coreceptors by a bispecific lentiviral vector. AB - BACKGROUND: RNA interference (RNAi) mediated by small interfering RNAs (siRNAs) has proved to be a highly effective gene silencing mechanism with great potential for HIV/AIDS gene therapy. Previous work with siRNAs against cellular coreceptors CXCR4 and CCR5 had shown that down regulation of these surface molecules could prevent HIV-1 entry and confer viral resistance. Since monospecific siRNAs targeting individual coreceptors are inadequate in protecting against both T cell tropic (X4) and monocyte tropic (R5) viral strains simultaneously, bispecific constructs with dual specificity are required. For effective long range therapy, the bispecific constructs need to be stably transduced into HIV-1 target cells via integrating viral vectors. RESULTS: To achieve this goal, lentiviral vectors incorporating both CXCR4 and CCR5 siRNAs of short hairpin design were constructed. The CXCR4 siRNA was driven by a U6 promoter whereas the CCR5 siRNA was driven by an H1 promoter. A CMV promoter driven EGFP reporter gene is also incorporated in the bispecific construct. High efficiency transduction into coreceptor expressing Magi and Ghost cell lines with a concomitant down regulation of respective coreceptors was achieved with lentiviral vectors. When the siRNA expressing transduced cells were challenged with X4 and R5 tropic HIV 1, they demonstrated marked viral resistance. HIV-1 resistance was also observed in bispecific lentiviral vector transduced primary PBMCs. CONCLUSIONS: Both CXCR4 and CCR5 coreceptors could be simultaneously targeted for down regulation by a single combinatorial lentiviral vector incorporating respective anti-coreceptor siRNAs. Stable down regulation of both the coreceptors protects cells against infection by both X4 and R5 tropic HIV-1. Stable down regulation of cellular molecules that aid in HIV-1 infection will be an effective strategy for long range HIV gene therapy. PMID- 15813992 TI - Glutamate transporter: an unexpected target for some antibiotics. AB - Glutamate transporter (GT) plays a major role in the mechanisms of glutamate homeostasis. Can this transporter system be a therapeutic target for glutamate mediated neurological disorders? In January's edition of Nature, Rothstein et al (2005) reports that the most commonly used class of antibiotics (beta-lactam antibiotics) such as ceftriaxone promoted the expression of GLT1 and demonstrated a functional role in both in vitro and in vivo models of glutamate neurotoxicity. These findings indicate that positive promoters of GT expression may have a unique role in neuroprotection through regulating GT expression. This is also encouraging in search for new pharmacological tools for pain management. PMID- 15813991 TI - Propofol suppresses synaptic responsiveness of somatosensory relay neurons to excitatory input by potentiating GABA(A) receptor chloride channels. AB - Propofol is a widely used intravenous general anesthetic. Propofol-induced unconsciousness in humans is associated with inhibition of thalamic activity evoked by somatosensory stimuli. However, the cellular mechanisms underlying the effects of propofol in thalamic circuits are largely unknown. We investigated the influence of propofol on synaptic responsiveness of thalamocortical relay neurons in the ventrobasal complex (VB) to excitatory input in mouse brain slices, using both current- and voltage-clamp recording techniques. Excitatory responses including EPSP temporal summation and action potential firing were evoked in VB neurons by electrical stimulation of corticothalamic fibers or pharmacological activation of glutamate receptors. Propofol (0.6 - 3 microM) suppressed temporal summation and spike firing in a concentration-dependent manner. The thalamocortical suppression was accompanied by a marked decrease in both EPSP amplitude and input resistance, indicating that a shunting mechanism was involved. The propofol-mediated thalamocortical suppression could be blocked by a GABAA receptor antagonist or chloride channel blocker, suggesting that postsynaptic GABAA receptors in VB neurons were involved in the shunting inhibition. GABAA receptor-mediated inhibitory postsynaptic currents (IPSCs) were evoked in VB neurons by electrical stimulation of the reticular thalamic nucleus. Propofol markedly increased amplitude, decay time, and charge transfer of GABAA IPSCs. The results demonstrated that shunting inhibition of thalamic somatosensory relay neurons by propofol at clinically relevant concentrations is primarily mediated through the potentiation of the GABAA receptor chloride channel-mediated conductance, and such inhibition may contribute to the impaired thalamic responses to sensory stimuli seen during propofol-induced anesthesia. PMID- 15813993 TI - Pavlovian fear memory induced by activation in the anterior cingulate cortex. AB - Identifying higher brain central region(s) that are responsible for the unpleasantness of pain is the focus of many recent studies. Here we show that direct stimulation of the anterior cingulate cortex (ACC) in mice produced fear like freezing responses and induced long-term fear memory, including contextual and auditory fear memory. Auditory fear memory required the activation of N methyl-D-aspartate (NMDA) receptors in the amygdala. To test the hypothesis that neuronal activity in the ACC contributes to unpleasantness, we injected a GABAA receptor agonist, muscimol bilaterally into the ACC. Both contextual and auditory memories induced by foot shock were blocked. Furthermore, activation of metabotropic glutamate receptors in the ACC enhanced behavioral escape responses in a noxious hot-plate as well as spinal nociceptive tail-flick reflex. Our results provide strong evidence that the excitatory activity in the ACC contribute to pain-related fear memory as well as descending facilitatory modulation of spinal nociception. PMID- 15813994 TI - Suppression of neutrophil accumulation in mice by cutaneous application of geranium essential oil. AB - BACKGROUND: Previous studies suggested that essential oils suppressed the adherence response of human neutrophils in vitro and that intraperitoneal application of geranium oil suppressed the neutrophil accumulation into peritoneal cavity in vivo. Usually, essential oils are applied through skin in aromatherapy in inflammatory symptoms. The purpose of this study is to assess the effects of cutaneous application of essential oils on the accumulation of neutrophils in inflammatory sites in skin of mice. METHODS: Inflammation with accumulation of inflammatory cells was induced by injection of curdlan, a (1-->3) beta-D-glucan in skin or peritoneal cavity of mice. Essential oils were applied cutaneously to the mice immediately and 3 hr after intradermal injection of curdlan. The skin with inflammatory lesion was cut off 6 hr after injection of curdlan, and the homogenates were used for myeloperoxidase (MPO: a marker enzyme of neutrophil granule) assay. RESULTS: The MPO activity of the skin lesion induced by curdlan was suppressed dose-dependently by cutaneous application of geranium oil. Other oils such as lavender, eucalyptus and tea tree oils also suppressed the activity, but their activities seemed weaker than geranium. Juniper oil didn't suppress the activity CONCLUSION: Cutaneous application of essential oils, especially geranium oil, can suppress the inflammatory symptoms with neutrophil accumulation and edema. PMID- 15813995 TI - Increased glutamate synaptic transmission in the nucleus raphe magnus neurons from morphine-tolerant rats. AB - Currently, opioid-based drugs are the most effective pain relievers that are widely used in the treatment of pain. However, the analgesic efficacy of opioids is significantly limited by the development of tolerance after repeated opioid administration. Glutamate receptors have been reported to critically participate in the development and maintenance of opioid tolerance, but the underlying mechanisms remain unclear. Using whole-cell voltage-clamp recordings in brainstem slices, the present study investigated chronic morphine-induced adaptations in glutamatergic synaptic transmission in neurons of the nucleus raphe magnus (NRM), a key supraspinal relay for pain modulation and opioid analgesia. Chronic morphine significantly increased glutamate synaptic transmission exclusively in one class of NRM cells that contains mu-opioid receptors in a morphine-tolerant state. The adenylyl cyclase activator forskolin and the cAMP analog 8-bromo-cAMP mimicked the chronic morphine effect in control neurons and their potency in enhancing the glutamate synaptic current was significantly increased in neurons from morphine-tolerant rats. MDL12330a, an adenylyl cyclase inhibitor, and H89, a protein kinase A (PKA) inhibitor, reversed the increase in glutamate synaptic transmission induced by chronic morphine. In addition, PMA, a phorbol ester activator of protein kinase C (PKC), also showed an increased potency in enhancing the glutamate synaptic current in these morphine-tolerant cells. The PKC inhibitor GF109203X attenuated the chronic morphine effect. Taken together, these results suggest that chronic morphine increases presynaptic glutamate release in mu receptor-containing NRM neurons in a morphine-tolerant state, and that the increased glutamate synaptic transmission appears to involve an upregulation of both the cAMP/PKA pathway and the PKC pathway. This glutamate mediated activation of these NRM neurons that are thought to facilitate spinal pain transmission may contribute to the reduced opioid analgesia during opioid tolerance. PMID- 15813996 TI - Worm sensation! PMID- 15813998 TI - Interaction of face and voice areas during speaker recognition. AB - Face and voice processing contribute to person recognition, but it remains unclear how the segregated specialized cortical modules interact. Using functional neuroimaging, we observed cross-modal responses to voices of familiar persons in the fusiform face area, as localized separately using visual stimuli. Voices of familiar persons only activated the face area during a task that emphasized speaker recognition over recognition of verbal content. Analyses of functional connectivity between cortical territories show that the fusiform face region is coupled with the superior temporal sulcus voice region during familiar speaker recognition, but not with any of the other cortical regions normally active in person recognition or in other tasks involving voices. These findings are relevant for models of the cognitive processes and neural circuitry involved in speaker recognition. They reveal that in the context of speaker recognition, the assessment of person familiarity does not necessarily engage supramodal cortical substrates but can result from the direct sharing of information between auditory voice and visual face regions. PMID- 15813997 TI - Controlling neuropathic pain by adeno-associated virus driven production of the anti-inflammatory cytokine, interleukin-10. AB - Despite many decades of drug development, effective therapies for neuropathic pain remain elusive. The recent recognition of spinal cord glia and glial pro inflammatory cytokines as important contributors to neuropathic pain suggests an alternative therapeutic strategy; that is, targeting glial activation or its downstream consequences. While several glial-selective drugs have been successful in controlling neuropathic pain in animal models, none are optimal for human use. Thus the aim of the present studies was to explore a novel approach for controlling neuropathic pain. Here, an adeno-associated viral (serotype II; AAV2) vector was created that encodes the anti-inflammatory cytokine, interleukin-10 (IL-10). This anti-inflammatory cytokine is known to suppress the production of pro-inflammatory cytokines. Upon intrathecal administration, this novel AAV2-IL 10 vector was successful in transiently preventing and reversing neuropathic pain. Intrathecal administration of an AAV2 vector encoding beta-galactosidase revealed that AAV2 preferentially infects meningeal cells surrounding the CSF space. Taken together, these data provide initial support that intrathecal gene therapy to drive the production of IL-10 may prove to be an efficacious treatment for neuropathic pain. PMID- 15813999 TI - Integration of visual and auditory information by superior temporal sulcus neurons responsive to the sight of actions. AB - Processing of complex visual stimuli comprising facial movements, hand actions, and body movements is known to occur in the superior temporal sulcus (STS) of humans and nonhuman primates. The STS is also thought to play a role in the integration of multimodal sensory input. We investigated whether STS neurons coding the sight of actions also integrated the sound of those actions. For 23% of neurons responsive to the sight of an action, the sound of that action significantly modulated the visual response. The sound of the action increased or decreased the visually evoked response for an equal number of neurons. In the neurons whose visual response was increased by the addition of sound (but not those neurons whose responses were decreased), the audiovisual integration was dependent upon the sound of the action matching the sight of the action. These results suggest that neurons in the STS form multisensory representations of observed actions. PMID- 15814001 TI - Prosody-driven sentence processing: an event-related brain potential study. AB - Four experiments systematically investigating the brain's response to the perception of sentences containing differing amounts of linguistic information are presented. Spoken language generally provides various levels of information for the interpretation of the incoming speech stream. Here, we focus on the processing of prosodic phrasing, especially on its interplay with phonemic, semantic, and syntactic information. An event-related brain potential (ERP) paradigm was chosen to record the on-line responses to the processing of sentences containing major prosodic boundaries. For the perception of these prosodic boundaries, the so-called closure positive shift (CPS) has been manifested as a reliable and replicable ERP component. It has mainly been shown to correlate to major intonational phrasing in spoken language. However, to define this component as exclusively relying on the prosodic information in the speech stream, it is necessary to systematically reduce the linguistic content of the stimulus material. This was done by creating quasi-natural sentence material with decreasing semantic, syntactic, and phonemic information (i. e., jabberwocky sentences, in which all content words were replaced by meaningless words; pseudoword sentences, in which all function and all content words are replaced by meaningless words; and delexicalized sentences, hummed intonation contour of a sentence removing all segmental content). The finding that a CPS was identified in all sentence types in correlation to the perception of their major intonational boundaries clearly indicates that this effect is driven purely by prosody. PMID- 15814000 TI - Role of the right and left hemispheres in recovery of function during treatment of intention in aphasia. AB - Two patients with residual nonfluent aphasia after ischemic stroke received an intention treatment that was designed to shift intention and language production mechanisms from the frontal lobe of the damaged left hemisphere to the right frontal lobe. Consistent with experimental hypotheses, the first patient showed improvement on the intention treatment but not on a similar attention treatment. In addition, in keeping with experimental hypotheses, the patient showed a shift of activity to right presupplementary motor area and the right lateral frontal lobe from pre- to post-intention treatment functional magnetic resonance imaging (fMRI) of language production. In contrast, the second patient showed improvement on both the intention and attention treatments. During pre-treatment fMRI, she already showed lateralization of intention and language production mechanisms to the right hemisphere that continued into post-intention treatment imaging. From pre- to post-treatment fMRI of language production, both patients demonstrated increased activity in the posterior perisylvian cortex, although this activity was lateralized to left-hemisphere language areas in the second but not the first patient. The fact that the first patient's lesion encompassed almost all of the dominant basal ganglia and thalamus whereas the second patient's lesion spared these structures suggests that the dominant basal ganglia could play a role in spontaneous reorganization of language production functions to the right hemisphere. Implications regarding the theoretical framework for the intention treatment are discussed. PMID- 15814002 TI - Second language interferes with word production in fluent bilinguals: brain potential and functional imaging evidence. AB - Bilingual individuals need effective mechanisms to prevent interference between their languages. Using event-related brain potentials (ERPs) and functional magnetic resonance imaging (fMRI), we present evidence for interference of phonological information from the nontarget language in German-Spanish bilinguals. A tacit picture-naming task was used in which bilinguals and monolinguals had to make speeded responses based on the first letter of the picture's name in the target language. In one condition, subjects were required to respond when the name began with a vowel and to withhold a response if it started with a consonant. Stimuli had been selected such that in half of the trials, the names in both languages necessitated the same response, whereas in the other half, responses were different for the two languages. For the bilinguals, the language in which the stimuli had to be named was changed after each block. Bilinguals showed phonological interference compared with monolingual performance, which was evident in their performance, ERPs, and fMRI patterns. Nonlanguagespecific brain areas such as the left middle prefrontal cortex were found to be crucial for the control of interference. PMID- 15814003 TI - Fusiform activation to animals is driven by the process, not the stimulus. AB - Previous studies have found that the lateral posterior fusiform gyri respond more robustly to pictures of animals than pictures of manmade objects and suggested that these regions encode the visual properties characteristic of animals. We suggest that such effects actually reflect processing demands arising when items with similar representations must be finely discriminated. In a positron emission tomography (PET) study of category verification with colored photographs of animals and vehicles, there was robust animal-specific activation in the lateral posterior fusiform gyri when stimuli were categorized at an intermediate level of specificity (e.g., dog or car). However, when the same photographs were categorized at a more specific level (e.g., Labrador or BMW), these regions responded equally strongly to animals and vehicles. We conclude that the lateral posterior fusiform does not encode domain-specific representations of animals or visual properties characteristic of animals. Instead, these regions are strongly activated whenever an item must be discriminated from many close visual or semantic competitors. Apparent category effects arise because, at an intermediate level of specificity, animals have more visual and semantic competitors than do artifacts. PMID- 15814004 TI - "Where to?" remote memory for spatial relations and landmark identity in former taxi drivers with Alzheimer's disease and encephalitis. AB - Recent research suggests that the hippocampus is not needed for the maintenance and recovery of extensively used environments learned long ago. Instead, a network of neocortical regions differentially supports memory for locationnavigation knowledge and visual appearance of well-known places. In this study, we present a patient, S. B., who was diagnosed with probable Alzheimer's disease long after retiring from his 40 years as a taxi driver in downtown Toronto, a place that he has visited rarely, if ever, in the last decade. His performance was compared to that of two other retired taxi drivers, L. R., who developed encephalitis after retirement, and I. L., who is without neurological illness, and a group of eight healthy control participants who were never taxi drivers but all of whom worked or lived in downtown Toronto until at least 10 years ago. Despite S. B.'s widespread atrophy, which has affected mainly his hippocampus and part of his occipitotemporal cortex, he performed at least as well as all other participants on remote memory tests of spatial location and mental navigation between well-known Toronto landmarks. Unlike the comparison populations, however, he was unable to discriminate between the appearances of landmarks that he had visited frequently in his many years as a taxi driver from unknown buildings. This profound deficit extended to famous world landmarks but not to famous faces and does not appear to be semantic in nature. These findings add further support to the claim that the hippocampus is not necessary for mental navigation of old environments and suggest that expertise is not sufficient to protect against landmark agnosia. PMID- 15814005 TI - Expectation and the vestibular control of balance. AB - Recent experiments have shown that the visual channel of balance control is susceptible to cognitive influence. When a subject is aware that an upcoming visual disturbance is likely to arise from an external agent, that is, movement of the visual environment, rather than from self-motion, the whole-body response is suppressed. Here we ask whether this is a principle that generalizes to the vestibular channel of balance control. We studied the whole-body response to a pure vestibular perturbation produced by galvanic vestibular stimulation (GVS; 0.5 mA for 3 sec). In the first experiment, subjects stood with vision occluded while stimuli were delivered either by the subject himself (self-triggered) or by the experimenter. For the latter, the stimulus was delivered either without warning (unpredictable) or at a fixed interval following an auditory cue (predictable). Results showed that GVS evoked a whole-body response that was not affected by whether the stimulus was self-triggered, predictable, or unpredictable. The same results were obtained in a second experiment in which subjects had access to visual information during vestibular stimulation. We conclude that the vestibular-evoked balance response is automatic and immune to knowledge of the source of the perturbation and its timing. We suggest the reason for this difference between visual and vestibular channels stems from a difference in their natural abilities to signal self-motion. The vestibular system responds to acceleration of the head in space and therefore always signals self-motion. Visual f low, on the other hand, is ambiguous in that it signals object motion and eye motion, as well as self-motion. PMID- 15814006 TI - Overlap in the functional neural systems involved in semantic and episodic memory retrieval. AB - Neuroimaging and neuropsychological data suggest that episodic and semantic memory may be mediated by distinct neural systems. However, an alternative perspective is that episodic and semantic memory represent different modes of processing within a single declarative memory system. To examine whether the multiple or the unitary system view better represents the data we conducted a network analysis using multivariate partial least squares (PLS ) activation analysis followed by covariance structural equation modeling (SEM) of positron emission tomography data obtained while healthy adults performed episodic and semantic verbal retrieval tasks. It is argued that if performance of episodic and semantic retrieval tasks are mediated by different memory systems, then there should differences in both regional activations and interregional correlations related to each type of retrieval task, respectively. The PLS results identified brain regions that were differentially active during episodic retrieval versus semantic retrieval. Regions that showed maximal differences in regional activity between episodic retrieval tasks were used to construct separate functional models for episodic and semantic retrieval. Omnibus tests of these functional models failed to find a significant difference across tasks for both functional models. The pattern of path coefficients for the episodic retrieval model were not different across tasks, nor were the path coefficients for the semantic retrieval model. The SEM results suggest that the same memory network/system was engaged across tasks, given the similarities in path coefficients. Therefore, activation differences between episodic and semantic retrieval may ref lect variation along a continuum of processing during task performance within the context of a single memory system. PMID- 15814007 TI - The neurophysiology of response competition: motor cortex activation and inhibition following subliminal response priming. AB - Some widely used tasks in cognitive neuroscience depend on the induction of a response conflict between choice alternatives, involving partial activation of the incorrect response before the correct response is emitted. Although such ''conflict tasks'' are often used to investigate frontal-lobe-based conflict monitoring processes, it is not known how response competition evolves in the motor cortex. To investigate the dynamics of motor cortex activation during response competition, we used a subliminal priming task that induced response competition while bypassing pre-response stage processing conflict. Analyses of movement-related EEG potentials supported an interaction between competing responses characterized by reciprocal inhibition. Inhibitory interactions between response channels contribute to the resolution of response conflict. However, the reciprocal inhibition at motor cortex level seemed to operate independent of higher level conflict-monitoring processes, which were relatively insensitive to response conflict induced by subliminal priming. These results elucidate how response conflict causes interference as well as the conditions under which frontal-lobe-based interference control processes are engaged. PMID- 15814008 TI - Right hemisphere activation of joke-related information: an event-related brain potential study. AB - Two studies tested the hypothesis that the right hemisphere engages in relatively coarse semantic coding that aids high-level language tasks such as joke comprehension. Scalprecorded event-related brain potentials (ERPs) were collected as healthy adults read probe words (CRAZY) preceded either by jokes or nonfunny controls ("Everyone had so much fun jumping into the swimming pool, we decided to put in a little water/platform"). Probes were related to the meaning of the jokes, but not the controls. In Experiment 1a, with central presentation, probes following jokes (related) elicited less negative ERPs 300-700 msec postonset (N400) than did probes following nonfunny controls (unrelated). This finding suggests related probes were primed by the jokes. In addition, unrelated probes elicited a larger anterior positivity 700-900 msec than did related, as irrelevant stimuli impacted control processes invoked by task demands. In Experiment 1b, probes (CRAZY) were preceded only by sentence-final words from jokes (water) or controls (platform). No ERP effects were observed in Experiment 1b, suggesting the N400 priming effect and the anterior positivity observed in Experiment 1a ref lect semantic activations at the discourse level. To assess hemispheric differences in semantic activations, in Experiment 2, ERPs were recorded as participants read probe words presented in their left and right visual fields (LVF and RVF, respectively). Probes elicited a smaller N400 component when preceded by jokes than controls. This N400 priming effect was larger with presentation to the LVF, suggesting joke-relevant information was more active in the right hemisphere. The anterior positivity was observed with RVF but not LVF presentation, suggesting an important role for the left hemisphere in controlled retrieval in language comprehension. PMID- 15814009 TI - Top-down enhancement and suppression of the magnitude and speed of neural activity. AB - Top-down modulation underlies our ability to selectively attend to relevant stimuli and to ignore irrelevant stimuli. Theories addressing neural mechanisms of top-down modulation are driven by studies that reveal increased magnitude of neural activity in response to directed attention, but are limited by a lack of data reporting modulation of neural processing speed, as well as comparisons with a perceptual (passive view) baseline necessary to evaluate the presence of enhancement and suppression. Utilizing functional MRI (fMRI) and event-related potential recordings (ERPs), we provide converging evidence that both the magnitude of neural activity and the speed of neural processing are modulated by top-down influences. Furthermore, both enhancement and suppression occur relative to a perceptual baseline depending on task instruction. These findings reveal the fine degree of influence that goal-directed attention exerts upon activity within the visual association cortex. We further document capacity limitations in top down enhancement corresponding with working memory performance deficits. PMID- 15814010 TI - Behavioral and electrophysiological evidence of a right hemisphere bias for the influence of negative emotion on higher cognition. AB - We examined how responses to aversive pictures affected performance and stimulus locked event-related potentials (ERPs) recorded during a demanding cognitive task. Numeric Stroop stimuli were brief ly presented to either left or right visual hemifield (LVF and RVF, respectively) after a centrally presented aversive or neutral picture from the International Affective Picture System. Subjects indicated whether a quantity value from each Stroop stimulus matched the preceding Stroop stimulus while passively viewing the pictures. After aversive pictures, responses were more accurate for LVF Stroops and less accurate for RVF Stroops. Early-latency extrastriate attention-dependent visual ERPs were enhanced for LVF Stroops. The N2 ERP was enhanced for LVF Stroops over the right frontal and parietal scalp sites. Slow potentials (300-800 msec) recorded over the frontal and parietal regions showed enhanced picture related modulation and amplitude for LVF Stroops. These results suggest that emotional responses to aversive pictures selectively facilitated right hemisphere processing during higher cognitive task performance. PMID- 15814011 TI - Theta responses are involved in lexical-semantic retrieval during language processing. AB - Oscillatory neuronal dynamics, observed in the human electroencephalogram (EEG) during language processing, have been related to the dynamic formation of functionally coherent networks that serve the role of integrating the different sources of information needed for understanding the linguistic input. To further explore the functional role of oscillatory synchrony during language processing, we quantified event-related EEG power changes induced by the presentation of open class (OC) words and closed-class (CC) words in a wide range of frequencies (from 1 to 30 Hz), while subjects read a short story. Word presentation induced three oscillatory components: a theta power increase (4-7 Hz), an alpha power decrease (10-12 Hz), and a beta power decrease (16-21 Hz). Whereas the alpha and beta responses showed mainly quantitative differences between the two word classes, the theta responses showed qualitative differences between OC words and CC words: A theta power increase was found over left temporal areas for OC words, but not for CC words. The left temporal theta increase may index the activation of a network involved in retrieving the lexical-semantic properties of the OC items. PMID- 15814012 TI - An epidemic averted through medical screening. PMID- 15814013 TI - Providing a quality service: direct referral from the cytology laboratory to the colposcopy clinic. AB - OBJECTIVES: To compare the time taken for the referral process and the accuracy of referrals before and after a process review and the introduction of a system of direct referral from the cytology laboratory to the colposcopy clinic. SETTING: The colposcopy service in a large teaching hospital in Teesside. METHODS: Data on time points within the referral process and smear histories were collected. Data on time points were obtained retrospectively from the case-notes from before the new system of referral ('pre' group) and from an electronic database after the changes ('post' group). Smear histories were retrieved from the cytology database. RESULTS: The overall time that patients waited from the time the smear was taken until the time they were seen in the colposcopy clinic was significantly reduced. The median time between smear and colposcopy decreased from 92.5 days (range 35-254 days) in the 'pre' group to 33 days (range 13-43 days) in the 'post' group (P=0.0001). The median time taken from the smear report being issued until the report arrived in the colposcopy clinic was 14 days (range 4-123 days) in the 'pre' group, compared with two days (range 0-17 days) in the 'post' group (P=0.0001). There was a significant reduction in the number of inaccurate referrals in the 'post' group compared with the 'pre' group (P=0.02). CONCLUSIONS: Direct referral significantly reduces the time patients wait for colposcopy appointments and improves the accuracy of referrals. PMID- 15814014 TI - Risk of invasive cervical cancer after Pap smears: the protective effect of multiple negatives. AB - OBJECTIVE: To determine the relationship between the number of initial negative Pap smears and risk of subsequent cervical cancer. DESIGN: A cohort study was conducted using data from the British Columbia Cervical Cancer Screening Program and British Columbia Cancer Registry. The analysis used a random sample (1%) of women aged 20-69 with Pap smears and all cases of invasive cervical cancer diagnosed between 1994 and 1999. Each negative screen defined the beginning of a screening interval and intervals longer than five years were truncated. The following variables were created for each interval: age at the beginning of the interval, interval length, previous cytological abnormality, previous cervical procedure and number of preceding consecutive negative screens. The relationship between these variables and risk of squamous cervical cancer was determined using survival analysis methods. RESULTS: A total of 388 cases of invasive cervical cancer (252 squamous) were included in the study from a study population of over 3.3 million Pap smears. The risk of invasive squamous cancer increased with time since the last negative screen, history of cytological abnormality and history of cervical therapeutic procedure. Risk was not significantly related to age (P=0.2) but was highest in women aged 30-49. Multiple consecutive negative pap smears were associated with reduced risk in women with a history of moderate atypia (P<0.0001), but not in women without a history (P=0.4). CONCLUSIONS: Multiple consecutive negative cytology was not associated with reduced risk of invasive cervical cancer in women with no history of cytological abnormality. PMID- 15814015 TI - Gender differences in utilization of colorectal cancer screening. AB - OBJECTIVES: To assess the demographic and psychological mediators of gender differences in uptake of flexible sigmoidoscopy (FS) screening for colorectal cancer. SETTING: A subsample (n=5462) from a large community trial of FS in the UK. METHODS: Men and women randomized to screening as part of the UK Flexible Sigmoidoscopy Trial were sent a postal questionnaire assessing demographic characteristics and attitudes to screening six months before their screening appointment. Attendance at screening was recorded by the screening centres. RESULTS: More men than women attended screening (73% versus 67%). The higher male attendance was partially explained by their lower levels of socioeconomic deprivation, higher levels of marital status and lower perceived barriers to screening. CONCLUSIONS: Contrary to expectations, men were more likely than women to attend FS screening. This was partially explained by socioeconomic and attitudinal differences to screening, but additional research is needed to understand the key aspects of FS screening that will maximize screening uptake in men and women. PMID- 15814016 TI - Colorectal cancer screening in Finland: details of the national screening programme implemented in Autumn 2004. AB - Colorectal cancer mortality can be reduced by repeated faecal occult blood (FOB) testing followed by colonoscopy for test positives. The object of this report is to describe how to launch a new screening programme in such a way that its effectiveness can be reliably evaluated. The programme is based on gradual expansion over time with individual-level randomization into screening or control arms among a target population aged 60-69 years in Finland. The target population will be sampled from the population register for invitees and controls by municipality and by birth cohort. The non-invited controls will gradually be screened only after the six-year implementation period. After 10 years, the programme covers the entire target population. The effects of screening will be evaluated, comparing the incidence of and mortality from colorectal cancer in those invited to screening with controls. The primary screening test is a biannual guaiac-based FOB test with three test cards for consecutive samples. In September-December 2004, around 5000 test-kits were sent to 22 piloting municipalities. In 2005, the programme expands both among municipalities and the target population, resulting in nearly 20,000 individual requests. The implementation of colorectal cancer screening in Finland in this way meets the criteria for a randomized controlled trial and the requirements for a public health programme. It allows unbiased research data to be collected while introducing the programme and may set an example for the introduction of all national screening programmes. PMID- 15814017 TI - Validity of process indicators of screening for breast cancer to predict mortality reduction. AB - The aim of the study was to empirically assess the acceptable levels of process indicators as described in the European Community Guidelines using materials from the mammography service screening programmes. The Finnish programme was evaluated for effectiveness with a prior estimate of 0.74 for RR in Finland and 0.81 in Helsinki. Hence, the Finnish programme was likely to be somewhat less effective in terms of reduction in mortality than implied on the basis of early randomized trials, but probably approaching the same level of effectiveness. Finland therefore provides background data on the applicability of the process indicators that are indicators of performance and surrogates for effectiveness. The performance data on 10 Finnish screening centres at subsequent screens were used. These centres invited 687,000 women aged 50-64 years in 1991-2000. The mean compliance was 93% and the corresponding recall rate was 2.3%. The benign to malignant biopsy ratio was 0.43:1. The average breast cancer detection rate was 0.36%, 2.1 compared with the background incidence. The proportion of screen detected stage II+ cancers was 26%. Most, but not all, of these process indicators met the desirable reference values of the European Community. The specific criteria of the European Community on stage distribution, rates of screen-detected cancers by stage and detection rate to background incidence may need reconsideration. PMID- 15814018 TI - Comparison of the validity and reliability of two image classification systems for the assessment of mammogram quality. AB - OBJECTIVE: To compare the reliability and validity of two classification systems used to evaluate the quality of mammograms: PGMI ('perfect', 'good', 'moderate' and 'inadequate') and EAR ('excellent', 'acceptable' and 'repeat'). SETTING: New South Wales (Australia) population-based mammography screening programme (BreastScreen NSW). METHODS: Thirty sets of mammograms were rated by 21 radiographers and an expert panel. PGMI and EAR criteria were used to assign ratings to the medio-lateral oblique (MLO) and cranio-caudal (CC) views for each set of films. Inter-observer reliability and criterion validity (compared with expert panel ratings) were assessed using mean weighted observed agreement and kappa statistics. RESULTS: Reliability: Kappa values for both classification systems were low (0.01-0.17). PGMI produced significantly higher values than EAR. Agreement between raters was higher using PGMI than EAR for the MLO view (77% versus 74%, P < 0.05), but was similar for the CC view. Dichotomized ratings ('acceptable' or 'needs repeating') did not improve reliability estimates. VALIDITY: Kappa values between raters and the reference standard were low for both classification systems (0.05-0.15). Agreement between raters and the reference standard was higher using PGMI than EAR for the MLO view (74% versus 63%), but was similar for the CC view. Dichotomized ratings of the MLO view showed slightly higher observer agreement. CONCLUSIONS: Both PGMI and EAR have poor reliability and validity in evaluating mammogram quality. EAR is not a suitable alternative to PGMI, which must be improved if it is to be useful. PMID- 15814019 TI - Ascertainment and evaluation of interval cancers in population-based mammography screening programmes: a collaborative study in four European centres. AB - OBJECTIVES: The purpose of the present study was to estimate the interval cancer (IC) rates in four population-based mammography screening programmes in four countries with different health-care environments, different access to cancer registry data, and different age groups of women invited. SETTING: The screening programmes in Coimbra (Portugal), Dublin (Ireland), Stockholm (Sweden), and Turin (Italy) participated in the study. METHODS: All cancer cases were searched for in cancer registries. IC rates and other outcome measures from the screening programmes were estimated and compared between the centres. Poisson regression model was used to estimate the proportional incidence based on IC rate in relation to expected total breast cancer incidence rate in the absence of screening. RESULTS: There was a more than tenfold difference in the number of invited women at the first round between the involved centres. The IC rates varied between 4.3 and 23.8 per 10,000 women screened. The levels of IC rates in relation to the estimated background incidence varied from 0.35 up to 0.46 depending on age groups involved in the programme, but did not differ significantly between three of the four involved centres. CONCLUSIONS: IC rates were quite similar between three of the four centres despite the differences in target population, invited ages, length of building-up of the programmes and different health-care organizations. Different access to complete cancer registry data is likely to explain the lower IC rates in the fourth centre. PMID- 15814020 TI - Comparing the performance of mammography screening in the USA and the UK. AB - To compare the performance of screening mammography in the USA and the UK, a consecutive sample of screening mammograms was obtained in women aged 50 and older from 1996 to 1999 who participated in the Breast Cancer Surveillance Consortium in the USA (n = 978,591) and the National Health Service Breast Cancer Screening Program in the UK (n = 3.94 million), including 6943 diagnosed with breast cancer within 12 months of screening. Recall rates were defined as the percentage of screening mammograms with a recommendation for further evaluation including diagnostic mammography, ultrasound, clinical examination or biopsy, and cancer detection rates including invasive cancer and ductal carcinoma in situ diagnosed within 12 months of a screening mammogram. All results were stratified by whether examinations were first or subsequent and adjusted to a standard age distribution. Among women who underwent a first screening mammogram, 13.3% of women in the USA versus 7.2% of women in the UK were recalled for further evaluation (relative risk for recall 1.9; 95% CI 1.8-1.9). For subsequent examinations recall rates were approximately 50% lower, but remained twice as high in the USA as in the UK. A similar percentage of women underwent biopsy in each setting, but rates of percutaneous biopsy were lower and rates of open surgical biopsy were higher in the USA. Women undergo screening approximately every 18 months in the USA and every 36 months in the UK. Based on a 20-year period of screening, the estimated percentage of women who would be recalled for additional testing was nearly threefold higher in the USA. The number of cancers detected was also higher in the USA (55 versus 43), and most of the increase was in the detection of small invasive and in situ cancers. The numbers of large cancers detected ( > 2 cm) were very similar between the two countries. Recall rates are approximately two to three times higher in the USA than in the UK. Importantly, despite less frequent screening in the USA, there are no substantial differences in the rates of detection of large cancers. Efforts to improve mammographic screening in the USA should target lowering the recall rate without reducing the cancer detection rate. PMID- 15814022 TI - Expression genomics and drug development: towards predictive pharmacology. AB - Expression genomics can be defined as the study of the dynamic transciptome and its regulatory elements. Technologies are available that can assess transcripts on a genome-wide scale over time and across many samples. This comprehensive and dynamic database is being used to decipher signalling pathways and to identify new biomarkers and targets. Biomarkers emerging from these studies have prognostic potential and can be used to predict therapeutic outcome. The multiplex nature of this approach not only telescopes the time to discovery, but also allows for detection of complex interactions. Taken together, these capabilities, if carefully used, can speed drug development, enhance the identification of potent drug combinations and identify patient populations that will benefit from these new drugs. PMID- 15814023 TI - The importance of experimental design in proteomic mass spectrometry experiments: some cautionary tales. AB - Proteomic expression patterns derived from mass spectrometry have been put forward as potential biomarkers for the early diagnosis of cancer and other diseases. This approach has generated much excitement and has led to a large number of new experiments and vast amounts of new data. The data, derived at great expense, can have very little value if careful attention is not paid to the experimental design and analysis. Using examples from surface-enhanced laser desorption/ionisation time-of-flight (SELDI-TOF) and matrix-assisted laser desorption-ionisation/time-of-flight (MALDI-TOF) experiments, we describe several experimental design issues that can corrupt a dataset. Fortunately, the problems we identify can be avoided if attention is paid to potential sources of bias before the experiment is run. With an appropriate experimental design, proteomics technology can be a useful tool for discovering important information relating protein expression to disease. PMID- 15814024 TI - The identification and functional characterisation of conserved regulatory elements in developmental genes. AB - Understanding the mechanisms that govern the expression of genomes is one of the major challenges of the post-genomic era. Phylogenetic footprinting, which identifies genomic regions under evolutionary constraints, has proven helpful in finding cis-regulatory elements of transcription; however, this method may not be applicable across all evolutionary distances and for all types of genes. Recent results from vertebrate comparisons indicate that strong conservation of cis regulatory regions may occur more frequently in developmental regulator genes. This paper reviews methods of identifying conserved regulatory elements of developmental genes by comparative genomics, including new attempts to detect conserved features beyond simple sequence similarities. The results obtained are outlined and the authors comment on their functional and evolutionary implications. Finally, an evaluation of currently available methods of characterising the function of presumed conserved regulatory regions is presented, and problems such as promoter compatibility, assigning distant elements to their cognate genes and multifunctionality of elements, discussed. PMID- 15814025 TI - DNA methylation, nucleosome formation and positioning. AB - Recent mapping of nucleosome positioning on several long gene regions subject to DNA methylation has identified instances of nucleosome repositioning by this base modification. The evidence for an effect of CpG methylation on nucleosome formation and positioning in chromatin is reviewed here in the context of the complex sequence-structure requirements of DNA wrapping around the histone octamer and the role of this epigenetic mark in gene repression. PMID- 15814026 TI - Functional genomics of protein kinases in plants. AB - Functional genomics has revolutionised the way that scientists approach biological questions, allowing for the comprehensive characterisation of the function of related proteins encoded in a genome. The sequencing of the genome of the model system Arabidopsis thaliana has enabled the beginning of functional genomics and the study of protein kinase families in plants. The large family of genes encoding protein kinases is a primary target of functional genomics studies in plants due to their importance in diverse physiological processes. This paper describes the functional genomics tools used to study the families of protein kinases in Arabidopsis, as well as progress in uncovering the functions of these proteins. PMID- 15814027 TI - Dissecting the ethylene pathway of Arabidopsis. AB - The plant hormone ethylene regulates growth, development and stress responses. In recent years, various genomic and proteomic approaches have been initiated to understand both the range of ethylene responses in the plant and the mechanism of signal transduction. Transcriptional profiling experiments reveal broad-ranging effects of ethylene upon gene regulation, with up to 7 per cent of the genes examined demonstrating a significant level of response in one study. Both transcriptional and post-transcriptional mechanisms regulate the expression of components within the ethylene signal transduction pathway. The importance of post-transcriptional regulation via the ubiquitin/proteasome-mediated degradation pathway is apparent in studies on the accumulation of ethylene insensitive 3 (EIN3), a key transcription factor in the pathway. Protein complexes also play a role in modulating ethylene signal transduction, with interactions between the ethylene receptors and the Raf-like kinase constitutive triple response-1 (CTR1) being required for ethylene perception at the endoplasmic reticulum. In this paper, recent developments in unravelling the transcriptional and post transcriptional regulation of the ethylene signalling and response pathways are considered, along with the latest developments in unravelling the biochemical mechanism behind ethylene perception. PMID- 15814028 TI - The dynamics of the proteome: strategies for measuring protein turnover on a proteome-wide scale. AB - Quantitative proteomics captures the steady-state amount of a protein in a cell but does not explain how a change in protein amount is manifest -- whether through a change in synthesis or a change in degradation. If we are to understand the changes in the proteome, we will need to define such processes. In this brief review, strategies for the determination of intracellular protein dynamics on a proteome-wide scale are discussed. PMID- 15814029 TI - Efficacies of mefloquine alone and of artesunate followed by mefloquine, for the treatment of uncomplicated, Plasmodium falciparum malaria in eastern Sudan. AB - In late 2003, the efficacies of mefloquine monotherapy and of an artesunate mefloquine combination, for the oral treatment of uncomplicated, Plasmodium falciparum malaria, were investigated and compared in New Halfa, in eastern Sudan. Of the patients who completed the 28 days of follow-up, 40 were treated only with single-dose mefloquine (at a dose of 25 mg/kg), and 38 with artesunate (at 4 mg/kg. day) for 3 days followed by single-dose mefloquine (at 15 mg/kg), given on the third day. Compared with those given the combination, the patients given mefloquine alone were more likely to suffer nausea, vomiting and dizziness (25.0% v. 2.6%; P=0.005) and to be found gametocytaemic (12.5% v. 0%; P=0.02) after treatment, and more likely to be found febrile (i.e. with a temperature >37.5 degrees C) on day 2 (25.0% v. 2.6%; P=0.005), although no patients were found febrile on day 3. Six of the patients--three (7.5%) of those given mefloquine only and three (7.9%) of those given the combination (P>0.05)- appeared to be treatment failures. Parasite genotyping indicated, however, that, although five of these six patients had true recrudescences, one (who had been treated with the combination) had been re-infected during the follow-up. The true frequencies of cure were therefore 92.5% after mefloquine alone and 94.7% after the combination (P>0.05). Thus, although the treatments appeared equally effective in clearing parasitaemias, the combination was better at clearing gametocytaemias and was less likely to cause adverse side-effects. It remains unclear why mefloquine given alone was almost 10-fold more likely to trigger adverse effects than treatment with a combination that contained the same drug. This may be a reflection of the different mefloquine doses and, for the patients given the combination, of the use of artesunate before the mefloquine treatment. PMID- 15814030 TI - ABO-blood-group types and protection against severe, Plasmodium falciparum malaria. AB - Although the ABO blood group of the human host has been reported to influence malarial infection, there have been few clinical observations on this effect. A hospital-based, comparative study was therefore performed to investigate the relationship between blood-group type and severe disease i nPlasmodium falciparum malaria. Overall, 243 cases of malaria (163 uncomplicated and 80 severe) and 65 patients with severe, non-malarial infections were studied. In terms of ABO-blood group composition, the patients with severe malaria were significantly different from the patients with the uncomplicated disease (P<0.001) and also from a population control described previously (P<0.0001). The patients with uncomplicated malaria or severe but non-malarial disease were, however, similar to the population control. The cases of severe malaria were significantly less likely to be of blood group O (P=0.0003), and significantly more likely to be of group AB (P<0.0001), than the patients with nonsevere malaria. It appears that individuals who are of blood-group O are relatively resistant to the severe disease caused by P. falciparum infection. PMID- 15814031 TI - Fatal visceral leishmaniasis, with massive bone-marrow infection, in an immunosuppressed but HIV-negative Spanish patient, after the initiation of treatment with meglumine antimoniate. AB - Although visceral leishmaniasis is often fatal in the developing world, Leishmania-attributable deaths in Europe are relatively rare and nowadays almost always linked to HIV infection. In Spain, however, a HIV-negative man with a history of chronic obstructive pulmonary disease and prednisone treatment was recently hospitalized because of hypotension and asthenia. Although the patient was afebrile, a bone-marrow aspirate, collected after thrombo- and leuco cytopenia had been observed, was found to contain huge numbers of amastigotes. A course of antileishmanial treatment with meglumine antimoniate was initiated but the patient went into refractory shock and died within 6 h. The significance of this case, in terms of the routine investigation and treatment of immunosuppressed patients who may have leishmaniasis, is discussed. PMID- 15814032 TI - The mechanism behind the antileishmanial effect of zinc sulphate. II. Effects on the enzymes of the parasites. AB - When used in vitro, zinc sulphate has a direct antileishmanial effect. To see if this effect involved the inhibition of the parasites' enzymes, extracts of the promastigotes and axenic amastigotes of Leishmania major (MHOM/IQ/93/MRC6) and L. tropica (MHOM/IQ/93/MRC2) were prepared. Zinc sulphate, at various concentrations, was then added to samples of these extracts before the activities, in the samples, of certain key enzymes of the Embden-Meyerhof pathway, hexose-monophosphate shunt and citric-acid cycle, and of two enzymes associated with virulence (protease and acid phosphatase), were determined. The zinc was found to inhibit every enzyme investigated, usually in a dose-dependent manner. Thus the direct antileishmanial effect of zinc may result, partially or entirely, from the inhibition of enzymes that are necessary for the parasites' carbohydrate metabolism and virulence. PMID- 15814033 TI - Rapid assessment of the geographical distribution of lymphatic filariasis in Uganda, by screening of schoolchildren for circulating filarial antigens. AB - To permit improvements in the targeting of control activities, the geographical distribution of lymphatic filariasis in Uganda was assessed by using a rapid immunochromatographic card test to check school-aged children for Wuchereria bancrofti-specific circulating filarial antigens (CFA). Survey sites were selected to represent the various ecological and topographical diversities in the country. Overall, 17,533 children from 76 sites were examined. CFA-positive cases were detected at 31 of the sites, with prevalences ranging from 0.4% to 30.7%. There appeared to be strikingly more lymphatic filariasis in the north of the country than in the south. The main focus was north of the Victoria Nile, where 27 (66%) of 41 sites had CFA-positive cases, often at high prevalences. Only four (11.4%) of the 35 sites south of the Victoria Nile had CFA-positive cases, and all four were along the western rift valley and had relatively low CFA prevalences. Geostatistical interpolation was used to create a map showing the geographical distribution of CFA prevalences in Uganda (by ordinary kriging), and to assess the population exposed to W. bancrofti transmission. Estimates based on population data from 2002 indicated that approximately 8.7 million people (35.3% of the national population) lived in areas where > 1% of the school-aged children were CFA-positive. CFA prevalences generally decreased with increasing altitude, and no CFA-positive cases were found at sites that were > 1300 m above sea level. Although it gives an under-estimate of the overall community prevalence (a fact that should be taken into account when interpreting the present results and comparing them with the results of other surveys), the screening of schoolchildren for CFA was found to be a simple and useful approach for mapping the geographical distribution of lymphatic filariasis. PMID- 15814035 TI - Refusal to take ivermectin: the associated 'risk' factors in Hoima district, Uganda. AB - Community-directed treatment with ivermectin (CDTI), for the control of onchocerciasis, was launched in Uganda in 1997. In East Africa, as in other endemic areas, the refusal to take the drug reduces treatment coverage and therefore poses a serious threat to attempts to eliminate onchocerciasis as a disease of public-health importance. In early 2003, an attempt was therefore made to determine the factors associated with refusal to take ivermectin during mass treatment in 2002, by interviewing the individuals, from three Ugandan villages with CDTI, who had been eligible to receive ivermectin. The subject's socio demographic characteristics, history of adverse reactions and participation in CDTI-related activities were explored, as potential 'risk' factors for refusing ivermectin in the treatment round in 2002. When odds ratios (OR) for refusal and their associated 95% confidence intervals (CI) were calculated, the villagers aged 18-35 years were found more likely to have received ivermectin in 2002 than their older counterparts (OR = 0.55; CI = 0.33-0.93). Individuals with an inter current medical condition were also more likely to have received ivermectin in 2002 than the other villagers (OR = 0.56; CI = 0.32-0.99). Worryingly, the ingestion of ivermectin at any time before the 2002 treatment round was strongly associated with refusal to take ivermectin in that round (OR = 7.72; CI = 2.00 29.81), although there was no evidence that the adverse effects of treatment were significantly reducing coverage. PMID- 15814034 TI - Significant decrease in the prevalence of Wuchereria bancrofti infection in anopheline mosquitoes following the addition of albendazole to annual, ivermectin based, mass treatments in Nigeria. AB - A prospective entomological survey was conducted in four sentinel villages in central Nigeria from 1999-2002, to assess the impact of annual, single-dose, mass drug administrations (MDA), with a combination of ivermectin and albendazole, on the transmission of Wuchereria bancrofti. As they were also endemic for human onchocerciasis, the four villages had received annual MDA based on ivermectin alone for 7 years prior to the addition of albendazole. Resting Anophelines gambiae s. l., An. funestus and Culex species were collected from 92 sequentially sampled households and dissected. Mosquitoes harbouring any larval stage of W. bancrofti were classified as 'infected', and those containing the third-stage larvae of the parasite were classified as 'infective'. Over the 41-month observation period, 4407 mosquitoes were captured and dissected, of which 64% were An. gambiae s. l., 34% An. funestus, and 1% Culex species. The baseline data, from dissections performed before the addition of albendazole to the MDA, showed high prevalences of mosquito infection (8.9%) and infectivity (2.9%), despite apparently good treatment coverages during the years of annual ivermectin monotherapy. Only the anopheline mosquitoes were found to harbour W. bancrofti larvae. After the third round of MDA with the ivermectin-albendazole combination, statistically significant decreases in the prevalences of mosquito infection (down to 0.6%) and infectivity (down to 0.4%) were observed (P<0.0001 for each). The combination of albendazole and ivermectin appears to be superior to ivermectin alone for reducing the frequency of W. bancrofti infection in mosquitoes. PMID- 15814036 TI - Infection with transfusion-transmitted virus (TTV) in humans and other primates in Venezuela. AB - Tranfusion-transmitted virus (TTV), a single-stranded circular DNA virus that chronically infects humans and other animals, displays a high degree of genetic diversity and was originally thought to be associated with hepatitis. The prevalences of TTV infection among different populations of humans and non-human primates from Venezuela have now been evaluated, using serum samples and three different detection tests. All three tests were PCR-based, one involving a hemi nested PCR and primers based on the N22 open-reading-frame-1 region (N22-PCR), another employing 55 cycles with primers from the more conserved untranslated region (UTR-PCR), and the other using a hemi-nested PCR with primers from the same region (HUTR-PCR). The overall prevalences of human infection appeared much higher with the HUTR-PCR (52%) than with the N22-PCR (13%) or the UTR-PCR (5%). When the products amplified by N22-PCR from 28 human isolates of TTV were sequenced, only two genotypes of the virus were detected. The non-human sera tested came from primates kept in a zoo in north-western Venezuela. TTV DNA was detected, by HUTR-PCR, in both of the chimpanzee sera tested but not in any of the sera from the 11 New-World primates or the other 12 Old-World primates that were investigated. The results, particularly those of the HUTR-PCR, indicate that TTV infection is common in Venezuela, especially in populations, such as many Amerindian groups, who live under poor sanitary conditions. Although TTV infection may be relatively rare among non-human primates from the New World, this will have to be investigated further, using many more samples collected throughout the Americas. PMID- 15814037 TI - Incrimination of Simulium thyolense (Diptera: Simuliidae) as the anthropophilic blackfly in the Thyolo focus of human onchocerciasis in Malawi. AB - The sibling species of the Simulium damnosum complex that are potential vectors in the foci of human onchocerciasis found in southern Malawi (the southern-most foci in Africa) were identified, using a combination of cytotaxonomic and molecular techniques. Simulium thyolense was found to be the most abundant species within the foci (representing 91% of the larvae collected), whilst S. kilibanum and cytoforms of the 'Ketaketa' subcomplex were the dominant members of the complex found outside of the foci. As all 188 biting female flies collected from the Thyolo, Mwanza and Mulanje foci were identified as S. thyolense, this species is probably the only significant vector of Onchocerca volvulus in the area, and the other Simulium species collected are assumed to be zoophilic. The results confirm the diagnostic value of the size of the amplicon(s) produced in PCR based on the sequences coding for the internal-transcribed-spacer region of the nuclear ribosomal DNA (ITS-1 rDNA), with a characteristic 340bp amplicon for S. thyolense. Chromosomal polymorphisms within S. thyolense indicate some degree of geographical isolation of the 'Thyolo-Mulanje' focus from the Mwanza focus. PMID- 15814038 TI - Relative abundances of sandfly species (Diptera: Phlebotominae) in two villages in the same area of Campeche, in southern Mexico. AB - Leishmania mexicana is the parasite causing most cases of human cutaneous leishmaniasis in southern Mexico, where Lutzomyia olmeca olmeca and Lu. cruciata are the most probable vectors. In the present study, sandflies were collected during one transmission season (November 2001-March 2002) in the village of La Guadalupe and the nearby village of Dos Naciones, in the southern Mexican county of Calakmul. Using Shannon traps, Disney traps and CDC light traps, 5983 sandflies (Brumptomyia and Lutzomyia) were caught. In Dos Naciones the numbers of Lu. panamensis caught in Shannon or CDC traps outnumbered those of the other sandfly species. In La Guadalupe, in contrast, the most abundant species in the collections made with Shannon or CDC traps was Lu. cruciata , followed by Lu. olmeca olmeca and Lu. deleoni. In both locations, the numbers of sandflies attracted to Shannon traps peaked between 18.00 and 22.00 hours. Given the abundance of Lu. olmeca olmeca in the collections made with Shannon and Disney traps (it was the only species caught in the latter), this species is probably the primary vector of Le. mexicana in Calakmul county. PMID- 15814039 TI - Experimental control of Biomphalaria pfeifferi, the intermediate host of Schistosoma mansoni, by the ampullariid snail Lanistes varicus. AB - The biological control of the snail hosts of the trematodes that cause human schistosomiasis appears to be a promising method for achieving sustainable reductions in the transmission of the parasites. The possibility of using the Ghanaian strain of an ampullariid snail, Lanistes varicus, for the biological control of the main snail host of Schistosoma mansoni , Biomphalaria pfeifferi, has now been investigated in laboratory-based experiments. Adult and 2-week-old L. varicus were found to feed voraciously on the egg masses and juveniles of B. pfeifferi (from the Tono irrigation canals in northern Ghana). When single L. varicus were exposed to 20-200 egg masses, they consumed all of the masses over 24 h (if adult) or about 50% of them over 4 days (if 2-week-old juveniles). The effect of the secretions of the ampullariid on the reproduction, growth and mortality of B. pfeifferi was also investigated, by maintaining the two snail species in the same aquarium but separated by nylon netting. The presence of L. varicus in the same aquarium reduced the number of egg masses produced by each B. pfeifferi, although, curiously, the presence of a single L. varicus in the aquarium appeared to have more of an impact, on the egg-mass deposition by 20 B. pfeifferi, than the presence of five or more of the ampullariids. It appears that, under laboratory conditions at least, the Ghanaian stain of L. varicus has the potential to limit populations of B. pfeifferi. PMID- 15814040 TI - The relative susceptibilities of juvenile and adult Bulinus globosus and Bulinus truncatus to the molluscicidal activities in the fruit of Ghanaian Blighia sapida, Blighia unijugata and Balanites aegyptiaca. AB - The relative susceptibilities of juvenile (2- to 3-week-old) and adult (5- to 7 week-old) Bulinus globosus and Bu. truncatus to the molluscicidal activities of aqueous preparations of the powdered, dried fruits of Blighia sapida, Bl. unijugata and Balanites aegyptiaca were investigated under laboratory conditions. For the extract prepared from the freeze-dried, semi-ripe fruits of Bl. sapida, the concentrations killing 95% of the snails after an exposure for 24 h (LC95) were, respectively, 232.7 and 161.2 ppm for the juveniles and adults of Bu. globosus and 187.6 and 140.2 ppm for the juveniles and adults of Bu. truncatus. The corresponding values for Bl. unijugata were 136.9 and 98.7 ppm for Bu. globosus and 132.2 and 98.5 ppm for Bu. truncatus, respectively. Extracts of the freeze-dried, ripe fruit of Ba. aegyptiaca were much more active, however, giving LC95 of just 16.9 and 19.7 ppm for the juveniles and adults of Bu. globosus, and 14.2 and 12.0 ppm for the juveniles and adults of Bu. truncatus, respectively. In general, similar results were obtained when the fruit were oven-dried rather than freeze-dried but the potency of Balanites extracts produced from oven-dried fruit was relatively low. That the juvenile snails appeared less susceptible than the adults to the molluscicidal activity in each Blighia extract is interesting. This observation may have implications for snail control, in terms of the optimal frequency, timing and concentration of molluscicides applied in a particular habitat. PMID- 15814041 TI - Antagonism of the anticoagulant effect of warfarin caused by the use of Commiphora molmol as a herbal medication: a case report. PMID- 15814042 TI - Globalisation, nutrition and health. PMID- 15814043 TI - Changing patterns of under- and over-nutrition in South African children-future risks of non-communicable diseases. AB - Under- and over-nutrition in children in KwaZulu-Natal, South Africa was investigated comparing data collected from primary school children in a rural district (643 children aged 8-11 years in 1994) with secondary data from the National Schools Study (16,179 children, 4-11 years in 1994), the Vitamin A Consultative Group Study (408 children, 2-5 years in 1994) and the Income Dynamics Study (1,593 children, 2-11 years in 1998). Stunting and wasting (WHO/NCHS guidelines) and overweight and obesity (International Obesity Task Force guidelines) were retrospectively analysed from these studies and compared in the children aged 4-5 and 8-11 years. There was moderate stunting in 10-25%, wasting in 1-6%, 5-24% were overweight and 1-10% obese. Girls in the National Schools Study (p<0.005) and in the primary datasets (p=0.02) had a significantly higher prevalence of overweight than boys; girls (1.4%) were also more obese than boys (0.9%) in the Schools Study (p=0.002), and the boys significantly more stunted (p<0.005) and wasted (p<0.005). An increasing prevalence of overweight and obesity was seen in both the 4-5- and 8-11-year age-groups. The finding that moderate stunting co-exists with overweight and obesity suggests that patterns of under- and over-nutrition in South African children are changing and might indicate the early stages of a complex nutritional transition. Action is required to prevent the future risk of non-communicable diseases. PMID- 15814044 TI - Comparative trial of short-course ofloxacin for uncomplicated typhoid fever in Vietnamese children. AB - An open, randomised comparison of 2 or 3 days of oral ofloxacin (10 mg/kg/day) for uncomplicated typhoid fever was conducted in 235 Vietnamese children. Multi drug-resistant Salmonella typhi was isolated from 182/202 (90%) children and 5/166 (3%) tested isolates were nalidixic acid-resistant (Na(R)). Eighty-nine of 116 children randomised to 2 days and 107/119 randomised to 3 days were blood culture-positive and eligible for analysis. There were 12 (13.5%) failures in the 2-day group (six clinical failures, four blood culture-positive post treatment, two relapses) compared with eight (7.5%) failures in the 3-day group (four clinical failures, one blood culture-positive post treatment, three relapses) (OR 1.9, 95% CI 0.7-5.5,p = 0.17). There were no significant differences in the mean (95% confidence interval) fever clearance times (h) [92 (82-102) vs 101 (93-110), p = 0.18] or duration of hospitalisation (d) [7.6 (7.2-8.1) vs 8.0 (7.6-8.4), p = 0.19] between the two groups. There was one failure in the four eligible children infected with an Na(R) isolate of S. typhi. No adverse events were attributable to the ofloxacin. These results extend previous observations on the efficacy of short courses of ofloxacin for children with uncomplicated multi-drug-resistant typhoid fever. PMID- 15814045 TI - Can clinical signs predict hypoxaemia in Papua New Guinean children with moderate and severe pneumonia? AB - Pulse oximetry was performed on 77 children admitted with acute lower respiratory tract infections (ALRI) to the children's ward in Port Moresby General Hospital, Papua New Guinea over a 4-month period in 2002. Clinical findings were correlated with different levels of hypoxaemia, <93%, <90% and <85%. Cyanosis, head nodding and drowsiness were good predictors of hypoxia but lacked sensitivity. Decisions to use oxygen based on these signs would therefore result in a significant number of children with hypoxia not receiving oxygen. Pulse oximetry is the best indicator of hypoxaemia in children with ALRI and, although relatively expensive, its use might be cost-effective in controlling oxygen requirements. PMID- 15814046 TI - Comparison of urine contamination rates using three different methods of collection: clean-catch, cotton wool pad and urine bag. AB - Collecting uncontaminated urine specimens from infants is difficult. Commonly, an adhesive urinecollecting bag is used, which is uncomfortable. This study determined bacterial contamination rates using three methods of urine collection sequentially on the same infant (without known urinary tract infection)-clean catch, cotton wool (sanitary) pad and urine bag. The study was undertaken in children under 3 years of age in the Institute of Maternal and Child Health of Pernambuco (IMIP), Recife, Brazil. Urine samples were analysed using phase contrast microscopy and routine culture. Culture of bacteria at any level was interpreted as a contaminated urine specimen. Cultures with > 10(5) colony forming units/ml of one species by all three collection methods were regarded as true urinary tract infection and these children were excluded. Altogether, 534 urine samples from 191 patients were analysed. Median age was 2 months (1 day-36 months) and 124 (65%) were boys. Twelve children (6.3%) were considered to have true urinary tract infection, three were indeterminate and in 16 one or more samples were missing and all were excluded from analysis. There were more missing samples using the clean-catch method (12%) than when using the bag (4%) or pad (4%). Seventy-six of 160 (47.5%) children had evidence of bacterial contamination. Clean-catch specimens showed the least contamination (14.7%) and rates were similar between pads (29%) and bags (26.6%) (kappa = 0.40). Urine contamination rates were similar for sanitary pads and urine bags and significantly higher than for clean-catch (p<0.01). However, pads were a simple, non-invasive and comfortable alternative to bags. PMID- 15814047 TI - Molecular epidemiology of Escherichia coli diarrhoea in children in Tehran. AB - From July to December 2003, four categories of diarrhoeagenic Escherichia coli were investigated in Tehranian children with acute diarrhoea. Stool specimens of children under 5 years of age with diarrhoea (n=200) and matched controls (n=200) without diarrhoea were studied for the presence of entero-aggregative (EAEC), enteropathogenic (EPEC), enterotoxigenic (ETEC) and Shiga toxin-producing (STEC) E. coli by PCR identification of six different genes of diarrhoeagenic E. coli. STEC isolates were typed by O157 and H7 antisera. EAEC was the most prevalent category and was found in 24% of patients with diarrhoea and 8% of controls (p<0.0001). ETEC was isolated in 15.5% of patients with diarrhoea but not in any controls ( p<0.0001), STEC in 15% of patients and 2% of controls (p<0.0001) and EPEC in 6% of patients and 5% of controls. Of 30 STEC isolates from patients with diarrhoea, seven were O157:H7 and 23 were non-O157:H7. PMID- 15814048 TI - Visceral childhood leishmaniasis in Kagizman, eastern Turkey: a new endemic area. AB - Visceral leishmaniasis is endemic in some areas of Turkey; however, it has not been reported previously from the eastern part of the country. We reviewed 21 cases of visceral leishmaniasis admitted from eastern Turkey, 60% of whom were from the same region, Kagizman. PMID- 15814049 TI - Duodenal intubation tube model for investigation of cholestasis. PMID- 15814050 TI - Community-acquired disseminated methicillin-resistant Staphylococcus aureus infection: case report and clinical implications. AB - A 6-year-old girl with community-acquired disseminated infection caused by methicillin-resistant Staphylococcus aureus (MRSA) is described. She had sepsis, meningo-encephalitis, pyomyositis, osteomyelitis, pericarditis and pulmonary embolisation caused by a multi-resistant strain of MRSA. Vancomycin is not routinely recommended as the first-line antimicrobial agent for suspected Staphylococcus aureus infection; however, it should be considered pending susceptibility results in patients presenting with severe sepsis in areas where the prevalence of MRSA is high. PMID- 15814051 TI - Aniridia associated with ptosis in three generations of the same family. AB - Aniridia is a spectrum of abnormalities of the iris anatomy, which range from a total absence of the iris to a mild stromal hypoplasia with normal pupil. Aniridia associated with ptosis in three generations of the same family is described. The cases fit the autosomal dominant familial aniridia (AN1) type, a genetic form of congenital aniridia characterised by isolated ocular defects. PMID- 15814052 TI - Neonatal adrenal abscess revisited: the importance of raised inflammatory markers. AB - Adrenal abscesses rarely occur in neonates and usually present with non-specific signs and symptoms. Prompt diagnosis requires an index of suspicion. We describe right-sided adrenal haemorrhage and abscess formation in a newborn with hypoxic ischaemic encephalopathy following maternal post-partum haemorrhage and sepsis with Escherichia coli and Enterococcus faecalis. Percutaneous drainage of the abscess identified an E. coli isolate identical to that in the mother plus Candida albicans. PMID- 15814053 TI - Bilateral retrobulbar neuritis associated with mumps. PMID- 15814054 TI - Editorial. PMID- 15814056 TI - Diabetes and the menopause. AB - As life expectancy increases, women are spending more time in the postmenopausal phase of life. Diabetes is one of the most common chronic diseases in the world and its prevalence is increasing. Type 2 diabetes mellitus is more common than type 1 (it accounts for 90% of all cases) and is most frequent in obese individuals over the age of 40 years. In this review, the main problems faced by postmenopausal diabetic women are examined, and hormone replacement therapy (HRT) in this group of women is discussed. HRT appears to decrease the incidence of type 2 diabetes mellitus and to improve glycaemic control; the results vary according to the type of HRT and the route of administration. HRT also improves lipid profiles and transdermal delivery seems to decrease triglyceride levels in particular. There are conflicting data on the effect of HRT on coronary heart disease (CHD); however, it may be beneficial in younger postmenopausal diabetic women. Cardioprotective treatment adjuncts (such as statins or low-dose aspirin) may be advised in diabetic women with CHD risk factors who require HRT. However, their prescription is currently not recommended solely for the possible prevention of cardiovascular disease. HRT may also protect women from osteoporosis in diabetes, especially in type 1 diabetes mellitus. It is recommended that the lowest possible effective dose is used. In postmenopausal diabetic women in whom HRT is not suitable, alternatives such as bisphosphonates may be employed. In these women, vasomotor symptoms can also be improved using drugs such as venlafaxine or gabapentin. Based on current data, we have proposed a regimen that could be used for women with diabetes. PMID- 15814057 TI - The use of bisphosphonates in breast cancer. AB - Bisphosphonates will become increasingly important in the management of patients with breast cancer. Currently, bisphosphonates are used to treat bone metastasis because they effectively relieve pain, prevent pathological fractures and treat hypercalcaemia of malignancy. Recent advances in systemic adjuvant therapies for breast cancer are improving survival but many treatments are detrimental to bone and can increase the risk of fracture. The monitoring of breast cancer patients at risk of developing osteoporosis will become increasingly important as survival times improve and more potent treatments are developed. Bisphosphonates may also play a role as an adjuvant therapy for the prevention of bone metastasis in high risk breast cancer patients. PMID- 15814058 TI - Advances in the treatment of endometrial cancer. AB - Endometrial cancer (EC) most commonly affects postmenopausal women. It is curable if treated early, but tumours with adverse histopathological features or at an advanced stage are associated with a high mortality rate. These cancers require a complex therapeutic approach, consisting of surgery, radiotherapy, chemotherapy and/or hormonal therapy. As one of the leading causes of death from malignancy in women, EC has been subject to intense clinical investigation. This article examines recent advances in the surgical treatment of the disease, such as sentinel lymph node sampling and total laparoscopic hysterectomy, as well as topics such as conservative treatment of EC for fertility preservation. Furthermore, new agents for EC treatment are presented, for example inhibitors of the mTOR pathway and the angiogenesis-inhibitor VEGF-trap. PMID- 15814059 TI - Which procedure for incontinence? AB - Approximately 16-29% of women will complain of incontinence at the menopause. A multidisciplinary approach to treatment is essential and surgery is usually reserved for women who have failed to improve sufficiently with conservative measures, such as pelvic floor exercises. The Burch colposuspension remains the 'gold standard' procedure for stress urinary incontinence, against which all newer procedures are compared. It is normally carried out under general anaesthesia and requires an abdominal approach, most often by a low transverse incision. Tension-free vaginal tape (TVT) is the most popular procedure at present and has largely replaced colposuspension in many units. However, there are concerns about both its long-term safety and the lack of data on efficacy beyond the first few years. Initial reports of transobturator tape suggest that it may have a similar efficacy to TVT but with a lower rate of serious complications; however, large comparative studies and longterm follow-up data are currently unavailable. Collagen and silicone injected around the urethra can be used in a selected group of patients under local anaesthetic but the success rates are disappointingly low. PMID- 15814060 TI - Vertebroplasty and kyphoplasty. AB - Vertebral compression fractures are common in elderly populations and in particular in postmenopausal women as a consequence of osteoporosis. Percutaneous vertebroplasty and kyphoplasty are minimally invasive procedures that are increasingly used to treat persistent or severe acute pain from these fractures. Vertebroplasty works by augmenting the weak osteopenic vertebrae with polymethylmethacrylate (PMMA) bone cement and thus preventing further microfractures and associated pain. The aim of kyphoplasty is pain relief combined with restoration of vertebral body height and reduction in kyphosis. This is achieved by 'expanding' the fractured vertebra with a balloon and then filling of the resultant cavity with PMMA cement. Vertebroplasty and kyphoplasty are undertaken under local and general anaesthesia, respectively. Both procedures have a very low complication rate if properly performed by well trained clinicians using appropriate cement and technique and highquality imaging. Patient selection and the selection of the level at which the percutaneous vertebroplasty is to be done are of utmost importance for maximal therapeutic benefit. Additional trials are required to establish conclusively the effectiveness of both procedures compared with conservative medical therapy and each other. PMID- 15814061 TI - Clinical nurse specialists and the psychosexual needs of patients with gynaecological cancer. AB - Due to improved survival rates many women have been left with chronic problems following treatment for a gynaecological cancer. These include psychosexual issues, premature menopause and infertility. Common sexual problems may result from low self-esteem, loss of femininity, vaginal stenosis and dryness. In the UK, clinical nurse specialists are recognized as key members of the multidisciplinary teams required to care for women diagnosed with a gynaecological cancer. Their role in providing information and support with special reference to psychosexual problems is examined. PMID- 15814062 TI - A burning mouth associated with the use of hormone replacement therapy. PMID- 15814063 TI - Extract signal out of noise. PMID- 15814064 TI - Impact of human population history on distributions of individual-level genetic distance. AB - Summaries of human genomic variation shed light on human evolution and provide a framework for biomedical research. Variation is often summarised in terms of one or a few statistics (eg F(ST) and gene diversity). Now that multilocus genotypes for hundreds of autosomal loci are available for thousands of individuals, new approaches are applicable. Recently, trees of individuals and other clustering approaches have demonstrated the power of an individual-focused analysis. We propose analysing the distributions of genetic distances between individuals. Each distribution, or common ancestry profile (CAP), is unique to an individual, and does not require a priori assignment of individuals to populations. Here, we consider a range of models of population history and, using coalescent simulation, reveal the potential insights gained from a set of CAPs. Information lies in the shapes of individual profiles--sometimes captured by variance of individual CAPs--and the variation across profiles. Analysis of short tandem repeat genotype data for over 1,000 individuals from 52 populations is consistent with dramatic differences in population histories across human groups. PMID- 15814065 TI - Cosmopolitan linkage disequilibrium maps. AB - Linkage maps have been invaluable for the positional cloning of many genes involved in severe human diseases. Standard genetic linkage maps have been constructed for this purpose from the Centre d'Etude du Polymorphisme Humain and other panels, and have been widely used. Now that attention has shifted towards identifying genes predisposing to common disorders using linkage disequilibrium (LD) and maps of single nucleotide polymorphisms (SNPs), it is of interest to consider a standard LD map which is somewhat analogous to the corresponding map for linkage. We have constructed and evaluated a cosmopolitan LD map by combining samples from a small number of populations using published data from a 10 megabase region on chromosome 20. In support of a pilot study, which examined a number of small genomic regions with a lower density of markers, we have found that a cosmopolitan map, which serves all populations when appropriately scaled, recovers 91 to 95 per cent of the information within population-specific maps. Recombination hot spots appear to have a dominant role in shaping patterns of LD. The success of the cosmopolitan map might be attributed to the co-localisation of hot spots in all populations. Although there must be finer scale differences between populations due to other processes (mutation, drift, selection), the results suggest that a whole-genome standard LD map would indeed be a useful resource for disease gene mapping. PMID- 15814066 TI - Trick or treat: the effect of placebo on the power of pharmacogenetic association studies. AB - The genetic mapping of drug-response traits is often characterised by a poor signal-to-noise ratio that is placebo related and which distinguishes pharmacogenetic association studies from classical case-control studies for disease susceptibility. The goal of this study was to evaluate the statistical power of candidate gene association studies under different pharmacogenetic scenarios, with special emphasis on the placebo effect. Genotype/phenotype data were simulated, mimicking samples from clinical trials, and response to the drug was modelled as a binary trait. Association was evaluated by a logistic regression model. Statistical power was estimated as a function of the number of single nucleotide polymorphisms (SNPs) genotyped, the frequency of the placebo 'response', the genotype relative risk (GRR) of the response polymorphism, the strategy for selecting SNPs for genotyping, the number of individuals in the trial and the ratio of placebo-treated to drug-treated patients. We show that: (i) the placebo 'response' strongly affects the statistical power of association studies--even a highly penetrant drug-response allele requires at least a 500 patient trial in order to reach 80 per cent power, several-fold more than the value estimated by standard tools that are not calibrated to pharmacogenetics; (ii) the power of a pharmacogenetic association study depends primarily on the penetrance of the response genotype and, when this penetrance is fixed, power decreases for larger placebo effects; (iii) power is dramatically increased when adding markers; (iv) an optimal study design includes a similar number of placebo and drug-treated patients; and (v) in this setting, straightforward haplotype analysis does not seem to have an advantage over single marker analysis. PMID- 15814068 TI - PBAT: a comprehensive software package for genome-wide association analysis of complex family-based studies. AB - The PBAT software package (v2.5) provides a unique set of tools for complex family-based association analysis at a genome-wide level. PBAT can handle nuclear families with missing parental genotypes, extended pedigrees with missing genotypic information, analysis of single nucleotide polymorphisms (SNPs), haplotype analysis, quantitative traits, multivariate/longitudinal data and time to onset phenotypes. The data analysis can be adjusted for covariates and gene/environment interactions. Haplotype-based features include sliding windows and the reconstruction of the haplotypes of the probands. PBAT's screening tools allow the user successfully to handle the multiple comparisons problem at a genome-wide level, even for 100,000 SNPs and more. Moreover, PBAT is computationally fast. A genome scan of 300,000 SNPs in 2,000 trios takes 4 central processing unit (CPU)-days. PBAT is available for Linux, Sun Solaris and Windows XP. PMID- 15814067 TI - A comprehensive literature review of haplotyping software and methods for use with unrelated individuals. AB - Interest in the assignment and frequency analysis of haplotypes in samples of unrelated individuals has increased immeasurably as a result of the emphasis placed on haplotype analyses by, for example, the International HapMap Project and related initiatives. Although there are many available computer programs for haplotype analysis applicable to samples of unrelated individuals, many of these programs have limitations and/or very specific uses. In this paper, the key features of available haplotype analysis software for use with unrelated individuals, as well as pooled DNA samples from unrelated individuals, are summarised. Programs for haplotype analysis were identified through keyword searches on PUBMED and various internet search engines, a review of citations from retrieved papers and personal communications, up to June 2004. Priority was given to functioning computer programs, rather than theoretical models and methods. The available software was considered in light of a number of factors: the algorithm(s) used, algorithm accuracy, assumptions, the accommodation of genotyping error, implementation of hypothesis testing, handling of missing data, software characteristics and web-based implementations. Review papers comparing specific methods and programs are also summarised. Forty-six haplotyping programs were identified and reviewed. The programs were divided into two groups: those designed for individual genotype data (a total of 43 programs) and those designed for use with pooled DNA samples (a total of three programs). The accuracy of programs using various criteria are assessed and the programs are categorised and discussed in light of: algorithm and method, accuracy, assumptions, genotyping error, hypothesis testing, missing data, software characteristics and web implementation. Many available programs have limitations (eg some cannot accommodate missing data) and/or are designed with specific tasks in mind (eg estimating haplotype frequencies rather than assigning most likely haplotypes to individuals). It is concluded that the selection of an appropriate haplotyping program for analysis purposes should be guided by what is known about the accuracy of estimation, as well as by the limitations and assumptions built into a program. PMID- 15814069 TI - 'A variant of uncertain significance' and the proliferation of human disease gene databases. AB - The rapid accumulation of mutation data has led to the creation of nearly 300 locus-specific mutation databases. These sites may contain a few dozen to almost 20,000 mutations for a given gene. Many of the mutations are uncharacterised and have no known effects on the gene product, the 'variant of uncertain significance'. Here, the statistics of mutation distribution are examined for six different gene databases: BRCA1 and BRCA2, haemoglobin-beta (HBB), HPRT1, CFTR and TP53. The percentage of all possible point mutations for a protein (the mutation space) is calculated for each gene and the question 'How much mutation data is enough?' is raised. PMID- 15814070 TI - Neuromyelitis Optica. AB - Neuromyelitis optica (Devic's syndrome) is an uncommon, idiopathic, demyelinating syndrome of the central nervous system that preferentially affects the optic nerves and spinal cord. It frequently is misdiagnosed as severe multiple sclerosis, but usually is readily distinguished from multiple sclerosis in fully developed cases because of its severity, typical magnetic resonance imaging (MRI) findings (normal brain MRI; longitudinally extensive lesions on spinal cord MRI), and cerebrospinal fluid analysis (polymorphonuclear pleocytosis and absence of oligoclonal banding). A serum autoantibody marker, NMO-IgG, is highly specific for the disorder. Most patients have relapsing disease, and natural history studies confirm early and severe disability. We treat acute myelitis and optic neuritis exacerbations with parenteral corticosteroids and use rescue plasmapheresis for severe, refractory attacks. Immunomodulatory drugs used for typical multiple sclerosis seem ineffective for relapse prevention. We recommend systemic immunosuppression, usually with azathioprine and oral corticosteroids, for most patients. Fulminant disease and breakthrough disease may respond to other forms of humoral immunotherapy such as rituximab. PMID- 15814071 TI - Gluten Ataxia and Post-Streptococcal Central Nervous System Syndromes: Emerging Immune-mediated Disorders of the Central Nervous System? AB - There is an "emerging concept" that central nervous system dysfunction can be caused by an aberrant immune response triggered by exogenous antigens such as the food allergen gluten or streptococcal infection. The hypothesis of a gluten sensitive ataxia remains unproven, but is worthy of consideration. The data in support of this hypothesis require critical review before any treatment recommendations can be formulated. The idea that anti-gliadin antibody seropositivity per se justifies the term "gluten sensitivity" is important because it offers potential therapeutic possibilities, including simple exclusion diets, for patients with anti-gliadin antibody-associated ataxia. Post streptococcal basal ganglia dysfunction has various manifestations, all of which fall into a relatively well-defined symptom complex or syndrome. Anti-basal ganglia antibodies that are associated with serologic evidence of recent streptococcal infection are a potential diagnostic marker for this group of disorders, which includes Sydenham's chorea (SC) as the prototype. More recently subjects with pediatric autoimmune neuropsychiatric disorders associated with streptococcal infection, Tourette's syndrome, obsessive-compulsive disorder and other movement disorders have been described in association with anti-basal ganglia antibodies. The apparent overlap between the clinical phenotype of SC, pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections, Tourette's syndrome, and obsessive-compulsive disorder suggests that they may represent one disease entity. The current working hypothesis is that antibodies induced in response to streptococcal infection cross-react with antigenic determinants in the basal ganglia resulting in basal ganglia dysfunction. Although the experimental evidence is incomplete, there is sufficient evidence to support immune-mediated basal ganglia dysfunction as an emerging clinical entity. This has important implications for the diagnosis and treatment of subjects with these disorders. The latter includes the judicious use of antibiotic prophylaxis and immunomodulatory therapies. Apart from the diagnosis and management of SC, no consensus exists regarding the diagnosis and management of the other clinical entities within this group of disorders. PMID- 15814072 TI - Treatment of Pediatric Multiple Sclerosis. AB - An increasing number of children and adolescents with multiple sclerosis (MS) are being identified and treated with disease-modifying therapies. Yet, there currently is little experience to guide clinicians. As more rapid diagnosis is made possible with magnetic resonance imaging, the number of pediatric MS patients requiring treatment will increase with time. This review draws on information from adult and pediatric neurology sources and summarizes current available data. Additional research is clearly needed. Nonetheless, there are several treatment principles: 1) establishing the diagnosis by differentiating MS from acute disseminated encephalomyelitis or recurrent acute disseminated encephalomyelitis; 2) emphasizing that there are parallels between adult and pediatric MS regarding the clinical presentation, disease course, and ability to tolerate therapy (the more extensive experience in adult MS can be applied to most pediatric cases); 3) explaining the importance of starting treatment early in the disease course and that these therapies seem well-tolerated in children; 4) providing reassurance that the family is not alone; as approximately 5% of the MS population has symptom onset before age 18 years (experience with pediatric MS is growing and centers with pediatric MS programs exist in other countries and are developing within the United States); and 5) reviewing symptom management and addressing issues such as academic performance that are unique to the pediatric MS subpopulation. PMID- 15814073 TI - Hemiballismus. AB - Hemiballismus is the most dramatic movement disorder seen in clinical practice. Its emergence points to a structural lesion or metabolic dysfunction in the region of the subthalamic nucleus, its afferent or efferent pathways, or interconnected deep brain nuclei, usually on the side contralateral to the movements. Any focal process may be to blame, but elderly sufferers generally have had vascular events, whereas the etiology is infectious or inflammatory in younger patients. Severe nonketotic hyperglycemia is another important cause of hemiballismus in the elderly. Hemiballismus patients require treatment both for the underlying etiology of the movement and for the movements themselves. There are no large controlled clinical trials to guide anti-ballismus therapy. However, dopamine receptor blocking agents have an established track record in suppressing choreic and ballistic movements, and are first-line agents for acute treatment. Standard neuroleptics such as haloperidol and perphenazine are started at low doses and titrated as tolerated until the movements are controlled. Atypical antipsychotics such as risperidone and clozapine have been used in small series and may have a reduced risk of extrapyramidal side effects. Catecholamine depleting agents such as reserpine and tetrabenazine may be considered when long term therapy is required. Other pharmacologic agents have met with varying success. The course of hemiballismus may be complicated by exhaustion, injury, or metabolic disorders, but with good supportive care, acute survival is good, and long-term survival reflects the prognosis of the underlying etiology. In time, the ballistic movements themselves tend to subside allowing withdrawal of drugs in many cases. When movements persist, stereotactic functional neurosurgical procedures may be considered in good surgical candidates. PMID- 15814074 TI - Tourette's Syndrome. AB - Tourette's syndrome is a childhood-onset neuropsychiatric disorder characterized by multiple motor and vocal tics, frequently accompanied by symptoms of obsessiveness and/or compulsiveness, anxiety, and behavioral impulsivity. Treatment of Tourette's syndrome symptoms should be considered when symptoms cause significant functional or social impairment or pain, as occurs with self injurious tics. Because comorbid psychiatric disorders, particularly attention deficit hyperactivity disorder (ADHD) and obsessive-compulsive disorder often are present, clinicians must work with affected persons and families and prioritize treatment targets based on the specific disorder-related impairment. Treatment with alpha-2 adrenergic agonists may reduce tics and improve ADHD symptoms. Effective treatment of ADHD, even with stimulant medications, in most cases does not exacerbate tics. Treatment with selective serotonin reuptake inhibitors may reduce obsessive-compulsive and anxiety symptoms, secondarily reducing tics. Neuroleptics and atypical antipsychotics may be used for severe tics, but the risk of neurologic side effects and weight gain is significantly higher. Habit reversal treatment shows promise as a nonpharmacologic intervention. Use of deep brain stimulation has produced benefit in three severely affected adults but should still be considered experimental. PMID- 15814075 TI - Myoclonus. AB - Myoclonus is defined as sudden, brief, shock-like involuntary movements affecting one or more muscles. The term encompasses a wide range of different physiologic and pathologic processes. When evaluating a patient with myoclonus, the first step is to identify the underlying etiology. Initial treatment should be directed against any underlying toxic or metabolic conditions. Next, targeted pharmacotherapy should be chosen, principally on the basis of the probable anatomical localization. Although treatment is initiated with a single agent, polytherapy usually is necessary to achieve adequate symptomatic control. The prognosis of myoclonus is highly variable, and largely depends on the underlying cause. PMID- 15814076 TI - Tardive Dystonia. AB - Tardive dyskinesia is a movement disorder that develops after exposure to dopamine receptor blocking agents. Less well-appreciated are other, more recently described tardive syndromes that are phenomenologically distinct from tardive dyskinesia and respond to different treatments. Patients may simultaneously have more than one tardive syndrome. Major subtypes of tardive syndromes include tardive dyskinesia, characterized by orobuccolingual, truncal, or appendicular, choreiform movements; tardive dystonia, characterized by sustained, stereotyped muscle spasms of a twisting or turning character; and tardive akathisia, characterized by an inner sense of restlessness or unease. The sensation often is unpleasant and may be accompanied by repetitive, purposeless movements (stereotypies), such as pacing. Less common tardive syndromes include tardive myoclonus, tardive tourettism, and tardive tremor. Tardive syndromes often are a source of great distress and disability to patients and may be permanent, despite discontinuing the responsible medication. Prevention, early detection, and prompt management are the major clinical focus. When a patient develops a tardive syndrome appropriate actions include 1) review of the primary diagnosis that prompted starting a dopamine receptor blocking agent; 2) characterization of the movement disorder(s); 3) where possible, discontinuation of dopamine blocking agent or replacement with a less potent alternative agent; 4) gradual withdrawl of the offending drug because some patients have an exacerbation of a tardive syndrome after abrupt withdrawal; and 5) assessment of the severity of symptoms and development of a treatment plan based on the phenomenology, with the goal of maximizing patient comfort and function. Although tardive dyskinesia typically develops after chronic exposure to dopamine receptor blocking agents, it, and other variants (such as tardive dystonia) can develop very rapidly after treatment. There seems to be no minimal safe duration of exposure for the development of a tardive syndrome. It is important to recognize that anti emetics, which are dopamine receptor blockers, such as prochlorperazine, promethazine and metoclopramide, can cause tardive syndromes. Clinicians should become familiar with antipsychotic agents that have a lower risk of causing tardive syndromes, such as clozapine, quetiapine, and olanzapine. We review treatment options for tardive dystonia. PMID- 15814077 TI - Deep Brain Stimulation for Dystonia. AB - Dystonia refers to movement disorders characterized by sustained muscle contractions that produce abnormal postures, twisting movements, and other abnormal involuntary movements. A spectrum of etiologies underlies the various dystonia syndromes, ranging from genetic conditions to brain injury. First-line therapy for dystonia consists of pharmacologic agents of several classes and, particularly for focal dystonia, chemodenervation therapy with botulinum toxin. Many patients with dystonia realize an inadequate response to those treatments, and for such patients whose symptoms are sufficiently troublesome, surgical treatment can be used to reduce symptoms and improve function. Previously, the ablative procedures of thalamotomy and pallidotomy were used, in which a permanent destructive lesion was made in the motor territory of the thalamus or the globus pallidus. More recently, the device-based therapy of deep brain stimulation (DBS) has emerged as the preferred surgical treatment for dystonia and other movement disorders for most patients who require operative intervention. DBS uses a surgically implanted brain lead connected to an implanted neurostimulator to deliver chronic, high- frequency electrical stimulation to one of several deep nuclei. For dystonia, stimulation directed at the globus pallidus internus has been the most thoroughly studied to date. Advantages of DBS include its relatively non-destructive nature, its adjustability and reversibility, and its capacity to be used bilaterally in a safe manner. Use of DBS to treat dystonia is a rapidly evolving area, and preliminary evidence suggests that primary dystonia linked to genetic mutation, other primary dystonias, and tardive dystonic syndromes respond most dramatically to treatment with DBS, whereas secondary dystonia tends to be less responsive. PMID- 15814078 TI - Preliminary evaluation of a broad-spectrum cognitive-behavioral group therapy for anxiety. AB - This study involved the development and outcome evaluation of a cognitive behavioral group treatment program for anxiety disorder using a randomized controlled design. This treatment program deviated from traditional anxiety disorder treatment protocols in that anxiety diagnosis was de-emphasized and treatment focused on shared common features across the anxiety disorders. Twenty three participants were recruited and randomly assigned to either immediate treatment or waitlist control conditions. Nine randomly assigned participants representing a range of anxiety diagnoses completed the 12-week group treatment, and were compared with 10 waitlist control participants who stayed in the study during the waitlist period. Participants in the two conditions were compared on change in diagnosis, diagnostic severity, self-report fear on ideographic measures, and self-report questionnaires of anxiety and state negative affect. Results were generally supportive of the efficacy of the treatment program. Compared to controls participants, those receiving treatment showed significantly greater improvement on diagnostic measures and ideographic fear-avoidance hierarchies. Data from self-report measures of anxiety and state negative affect were less supportive of the treatment efficacy. Implications for emerging conceptualizations of anxiety disorders, as well as implications for treatment and treatment dissemination, are discussed. PMID- 15814079 TI - Differentiating symptoms of social anxiety and depression in adults with social anxiety disorder. AB - Although studies have suggested a strong overlap between social anxiety disorder and depression, this is the first study to examine the ability of commonly used measures to differentiate symptoms of these disorders in a sample of clients with social anxiety disorder. Structural equation modeling revealed that commonly used measures of social anxiety and depression can differentiate the two constructs, rather than simply reflecting a single construct of overall distress. Logistic regression analyses indicated that scores on depressive symptom measures could predict which socially anxious clients met criteria for a comorbid depressive disorder. PMID- 15814080 TI - Implicit and explicit memory processes in panic patients as reflected in behavioral and electrophysiological measures. AB - Implicit and explicit memory processes for panic-relevant and neutral word stimuli were examined in 16 panic patients and 16 healthy participants matched for sex, age, and education, using behavioral and electrophysiological measures. In the study phase, panic-relevant and neutral words were presented and the level of processing was varied by requiring either shallow (orthographic) or deeper (syntactic) processing. Implicit memory was tested with a lexical decision task, explicit memory with a recognition task. Panic patients and healthy participants did not differ in behavioral (response time) or event-related brain potential (ERP) measures of implicit memory. However, panic patients deviated from healthy participants in the recognition test, an explicit memory test. Although recognition of panic words was overall worse compared to neutral words, panic patients compared to healthy participants exhibited enhanced discrimination scores and faster reaction times for panic words. The level of processing manipulation had comparable effects on patients and healthy participants. While neither behavioral nor electrophysiological measures provided evidence for an implicit memory bias in panic patients, behavioral measures confirmed an explicit memory bias in panic patients for panic-relevant stimuli. PMID- 15814082 TI - Memory-emotional interactions as revealed by fear generalization in animal fearful individuals. AB - Discriminative fear learning and fear generalization were examined in animal fearful individuals and in control participants. Electrical shocks were administered contingent upon discriminative pictures of spiders or snakes, respectively, in a generalization-after-discrimination paradigm. Neither discriminative fear learning nor extinction was affected by the individual fear status of the animal categories. Novel feared stimuli, which resembled discriminative stimuli, were treated as more shock predictive than novel non feared stimuli during generalization testing. Neither preparedness theory nor selective sensitization theory was capable to account for these observations. The findings are commensurable with the hypothesis that phobic fear interferes with the retrieval of memory traces. PMID- 15814081 TI - Stability and discriminative power of the Young Schema-Questionnaire in a Dutch clinical versus non-clinical population. AB - In this study the temporal stability and general discriminative and classifying ability of the Young Schema-Questionnaire (YSQ) was examined in a clinical and non-clinical sample. To be able to cross-validate on the variables, two parallel subtests, drawn from the YSQ item pool, were utilized. Results suggest adequate rank order stability. However, mean scores tended to drop systematically over time, most likely caused by transient error. Therefore, to assess progress in therapy, the alternate utilization of the parallel parts is advised. Findings from discriminant analysis suggest high sensitivity of the YSQ and its subscales in predicting the presence or absence of psychopathology. PMID- 15814083 TI - Biological activity of for-Met-Leu-Phe-OMe analogs: relevant substitutions specifically trigger killing mechanisms in human neutrophils. AB - Two analogs of the prototypical peptide for-Met-Leu-Phe-OMe (fMLP-OMe), for-Gln Tyr-Phe-OMe (1) and for-Gln-Tyr-Tyr-OMe (2), carrying unusual hydrophilic residues, were synthesized in order to investigate whether they provoked specific biological responses, as well as intracellular calcium mobilization, in human neutrophils. Whereas neither compound stimulates chemotaxis, both are able to elicit lysosomal enzyme production. However compound 1 is able to trigger copious superoxide anion production while compound 2 only elicits minor superoxide anion production. In binding experiments on formylpeptide receptors, the newly synthesized compounds for-Gln-Tyr-Phe-OMe (1) and for-Gln-Tyr-Tyr-OMe (2) showed affinity values in the micromolar range. These derivatives demonstrate inability to find a positive contribute from single substitutions. A very important result of this research is the evidence of the ability of the formyl group alone to trigger the primary target of the human neutrophil activity, i.e. killing mechanisms, by activating the specific receptor conformation. PMID- 15814084 TI - Cyclic AMP-independent activation of CYP3A4 gene expression by forskolin. AB - Forskolin and cAMP have been shown to have paradoxical effects in the regulation of expression levels of mRNA of cytochrome P450 3A (CYP3A) family members. We demonstrate in this study that forskolin upregulated the promoter for CYP3A4 independent of its ability to increase cAMP levels. This activity was explained showing forskolin directly activated the pregnane-X-receptor, a known regulator of CYP3A genes. PMID- 15814085 TI - Isoallopregnanolone; an antagonist to the anaesthetic effect of allopregnanolone in male rats. AB - The interaction of isoallopregnanolone (3 beta-OH-5 alpha-pregnan-20-one) on allopregnanolone (3 alpha-OH-5 alpha-pregnan-20-one) induced anaesthesia was studied in male rats using burst suppression of 1 s ("silent second") with an electroencephalographic-threshold method. The i.v. administration of isoallopregnanolone was varied in relation to induction of "silent second". Pre treatment with isoallopregnanolone (12.5-50 mg/kg iv) 2 min prior to the threshold test gave an increase in the threshold dose of allopregnanolone (ANOVA df(3;36), F=13.61, P<0.001), which was dose dependent (r=0.73, b [slope]=0.08, df=38, P<0.001). After isoallopregnanolone pre-treatment, but not in the controls, anaesthesia time was positively related to the dose of allopregnanolone (r=0.52, b=1.72, df=28, P<0.01). Anaesthesia times were not influenced by a corresponding administration of isoallopregnanolone immediately after induction of "silent second". When allopregnanolone and isoallopregnanolone were infused together at molar ratios of 1:1, 1:1.23, 1:1.43, a linear increase of the threshold doses of allopregnanolone was seen in relation to the dose of isoallopregnanolone (r=0.86, b=0.40, df=8, P<0.01). Thus isoallopregnanolone can antagonise the anaesthetic action of allopregnanolone. PMID- 15814086 TI - Delta-opioid receptor agonist SNC80 induces peripheral antinociception via activation of ATP-sensitive K+ channels. AB - We investigated the effect of several K+ channel blockers on the antinociception induced by delta-opioid receptor agonist SNC80 using the paw pressure test, in which pain sensitivity is increased by an intraplantar injection (2 microg) of prostaglandin E2 (PGE2). Administration of SNC80 (20, 40 and 80 microg/paw) caused a decrease in the hyperalgesia induced by PGE2, in a dose-dependent manner. The possibility of higher dose of SNC80 (80 microg) causing a central or systemic effect was excluded since administration of the drug into the contralateral paw did not elicit antinociception in the right paw. Specific blockers of ATP-sensitive K+ channels, glibenclamide (20, 40 and 80 microg/paw) and tolbutamide (40, 80 and 160 microg/paw), antagonized the peripheral antinociception induced by SNC80 (80 microg). On the other hand, charybdotoxin (2 microg/paw), a large-conductance blocker of Ca(2+)-activated K+ channels, and dequalinium (50 microg/paw), a small conductance selective blocker of Ca(2+) activated K+ channels, did not modify the effect of SNC80. This effect also remained unaffected by intraplantar administration of the voltage-dependent K+ channel blockers tetraethylammonium (30 microg/paw) and 4-aminopyridine (10 microg/paw), and of a non-specific K+ channel blocker, cesium (500 microg/paw). This study provides evidence that the peripheral antinociceptive effect of SNC80 result from the activation of ATP-sensitive K+ channels, and the other K+ channels are not involved. PMID- 15814087 TI - Brain prostanoid TP receptor-mediated adrenal noradrenaline secretion and EP3 receptor-mediated sympathetic noradrenaline release in rats. AB - Sympathetic nerves release noradrenaline, whereas adrenal medullary chromaffin cells secrete noradrenaline and adrenaline. Therefore, plasma noradrenaline reflects the secretion from adrenal medulla in addition to the release from sympathetic nerves, however the exact mechanisms of adrenal noradrenaline secretion remain to be elucidated. The present study was designated to characterize the source of plasma noradrenaline induced by intracerebroventricularly (i.c.v.) administered bombesin and prostaglandin E2 in urethane-anesthetized rats. Bombesin (1.0 nmol/animal, i.c.v.) elevated plasma noradrenaline and adrenaline, while prostaglandin E2 (0.3 nmol/animal, i.c.v.) elevated only plasma noradrenaline. The bombesin-induced elevations of both catecholamines were attenuated by pretreatments with furegrelate (an inhibitor of thromboxane A2 synthase) [250 and 500 microg (0.9 and 1.8 micromol)/animal, i.c.v.)] and [(+)-S-145] [(+)-(1R,2R,3S,4S)-(5Z)-7-(3-[4-3H]-phenylsulphonyl aminobicyclo[2.2.1]hept-2-yl)hept-5-enoic acid sodium salt] (an antagonist of prostanoid TP receptors) [100 and 250 microg (250 and 625 nmol)/animal)], and abolished by acute bilateral adrenalectomy. On the other hand, the prostaglandin E2-induced elevation of plasma noradrenaline was not influenced by acute bilateral adrenalectomy. These results suggest that adrenal noradrenaline secretion and sympathetic noradrenaline release are mediated by differential central mechanisms; brain prostanoid TP receptors activated by bombesin are involved in the adrenal noradrenaline secretion, while brain prostanoid EP (probably EP3) receptors activated by prostaglandin E2 are involved in the sympathetic noradrenaline release in rats. Brain prostanoid TP receptors activated by bombesin are also involved in the adrenal adrenaline secretion. PMID- 15814088 TI - Serotonin reuptake inhibitors attenuate morphine withdrawal syndrome in neonatal rats passively exposed to morphine. AB - Previous investigations had shown that inhibitor of serotonin reuptake transporter (SERT) could attenuate morphine withdrawal syndrome in adult animals. In the present study, we determined whether postnatal injection of serotonin reuptake inhibitors, fluoxetine, clomipramine, or citalopram, is able to attenuate the expression of the naloxone-precipitated morphine withdrawal syndrome in 5-day-old neonatal Sprauge-Dawley rats born to dams rat that received morphine injection since a week before mating till 5 days after delivery. Withdrawal syndrome of morphine, manifested as frequent abdominal stretching and yawning, was generated by injection of naloxone on postnatal day 5. Pre-injection with fluoxetine, clomipramine, or citalopram, significantly attenuated the naloxone-precipitated syndrome in a dose-dependent manner without apparent side effect. The rank order of inhibitory potency is citalopram=clomipramine>fluoxetine. This result suggests that inhibitor of SERT may be of potential in treating neonatal morphine withdrawal syndrome. PMID- 15814089 TI - Dual effects of bryostatin-1 on spatial memory and depression. AB - Dementia and depression are clinical symptoms commonly associated in patients. Emerging evidence suggests that the two diseases share many profiles in their development and underlying neural/molecular mechanisms. Thus, interest is raised in developing new classes of antidepressant agents with activity of cognitive enhancement. Here, we show that bryostatin-1, a protein kinase C substrate activator, at bilateral intracerebroventricular doses of 0.64 or 2 pmol/site, significantly enhanced learning and memory of rats in a spatial water maze task. When applied at the doses at which it exhibits memory-enhancing activity, bryostatin-1 showed a significant antidepressant activity, as determined in an open space swim test. Both effects were not observed when a smaller dose was administered and were largely eliminated by co-administration of 1-(5 isoquinolinesulfonyl)-2-methylpiperazine (H-7), a protein kinase C inhibitor. These results support the hypothesis that memory processing and mood regulation share common neural mechanisms. Restoring impaired mood regulation with antidepressant agents that also exhibit memory-enhancing activity may represent one of the new strategies in the fight against depression associated with memory impairments. PMID- 15814090 TI - Differential effects of human ether-a-go-go-related gene (HERG) blocking agents on QT duration variability in conscious dogs. AB - The effects of drugs that inhibit human ether-a-go-go-related gene (HERG) related cardiac potassium channels on the variability of QT duration as a sign of repolarisation instability were evaluated in conscious telemetered dogs. QT duration variability was determined using a beat-to-beat analysis before and after the infusions of HERG channel blocking agents. Variability was evaluated as increased mean width (P(width)) and length (P(length)) of Poincare plots of 100 consecutive beats. As HERG channel blockers which are associated with arrhythmias of the torsades de pointes (TdP) type, dofetilide and sotalol were infused. Verapamil was used as an HERG channel blocker that is not associated with TdP. Dofetilide (0.01 and 0.03 mg/kg) dose-dependently prolonged QT(c) duration (12% and 16%). Dofetilide also induced an increase of QT variability that reached statistical significance for P(length) at the higher dose (64%). A dose of 3 mg/kg sotalol neither prolonged QT(c) duration nor QT duration variability. In contrast, at 10 mg/kg sotalol prolonged QT(c) duration (15%) and increased P(length) (33%). Doses of 0.1 and 0.3 mg/kg verapamil did not increase QT(c) duration nor QT time variability. QT duration variability in conscious dogs may be a useful preclinical marker to discriminate pro-arrhythmogenic and non arrhythmogenic activities of HERG blocking agents. PMID- 15814091 TI - Effects of solifenacin succinate (YM905) on detrusor overactivity in conscious cerebral infarcted rats. AB - Solifenacin succinate [YM905, (+)-(1S,3'R)-quinuclidin-3'-yl 1-phenyl-1,2,3,4 tetrahydroisoquinoline-2-carboxylate monosuccinate] is a novel muscarinic receptor antagonist. We examined the effects of solifenacin and two other muscarinic receptor antagonists, tolterodine and propiverine, on detrusor overactivity in cerebral infarcted rats. Evaluation was done under conscious conditions using cystometry 1 day after middle cerebral artery occlusion. The cerebral infarcted rats showed decreases in bladder capacity and voided volume and an increase in residual volume, but no change in micturition pressure. Solifenacin increased bladder capacity and voided volume at doses of 0.03 mg/kg i.v. or more. Tolterodine increased bladder capacity and voided volume at 0.03 and 0.1 mg/kg i.v., while propiverine increased bladder capacity and voided volume at 1 mg/kg i.v. and at 0.3 and 1 mg/kg i.v., respectively. In contrast, none of the three drugs affected residual volume or micturition pressure. These results suggest that solifenacin may improve detrusor overactivity without causing urinary retention and may be a promising drug in the treatment of patients with overactive bladder syndrome. PMID- 15814092 TI - The epidermal growth factor-pathway is not involved in down-regulation of Ca(2+) induced Cl- secretion in rat distal colon. AB - Ca(2+)-dependent secretagogues such as carbachol induce a transient Cl- secretion followed by long-lasting inhibition (run-down) of secretion. In the colonic tumour cell line, T84, epidermal growth factor (EGF) inhibits Ca(2+)-dependent secretion, whereas antagonists of the EGF-signalling pathway slow down its run down. The aim of the present study was to investigate whether a similar mechanism underlies the down-regulation of carbachol-induced Cl- secretion measured as change in short-circuit current (I(sc)) in a native intestinal epithelium, i.e. rat distal colon. In contrast to the colonic tumour cell line, EGF (1-100 microg/l) induced a transient secretory I(sc) and did not interfere with a subsequent administration of carbachol. Pretreatment with inhibitors of enzymes involved in the signalling cascade induced by EGF, i.e. tyrphostin AG1478, an inhibitor of the EGF receptor protein tyrosine kinase, PD 98059, an inhibitor of MAP kinase, and wortmannin, a blocker of the phosphatidylinositol-3-kinase, did also not affect the action of carbachol on transepithelial I(sc). In order to investigate potential effects of these inhibitors on apical Cl- channels, the basolateral membrane was depolarized and a Cl- current across the apical membrane was driven by a Cl- gradient. Under these conditions, carbachol evoked a transient increase in I(sc), caused by the stimulation of Ca(2+)-dependent Cl- channels, followed by a long-lasting down-regulation of apical Cl- conductance leading to a decrease in I(sc). All blockers of the EGF-signalling pathway tested did not interfere with the action of carbachol at the apical membrane. Consequently, the EGF-pathway seems not to be involved in the down-regulation of Ca(2+)-dependent Cl- secretion across rat colon. PMID- 15814093 TI - Impaired verbal short-term memory in Down syndrome reflects a capacity limitation rather than atypically rapid forgetting. AB - Individuals with Down syndrome suffer from relatively poor verbal short-term memory. Recent work has indicated that this deficit is not caused by problems of audition, speech, or articulatory rehearsal within the phonological loop component of Baddeley and Hitch's working memory model. Given this, two experiments were conducted to investigate whether abnormally rapid decay underlies the deficit. In a first experiment, we attempted to vary the time available for decay using a modified serial recall procedure that had both verbal and visuospatial conditions. No evidence was found to suggest that forgetting is abnormally rapid in phonological memory in Down syndrome, but a selective phonological memory deficit was indicated. A second experiment further investigated possible problems of decay in phonological memory, restricted to item information. The results indicated that individuals with Down syndrome do not show atypically rapid item forgetting from phonological memory but may have a limited-capacity verbal short-term memory system. PMID- 15814094 TI - The strategic regulation of children's memory performance and suggestibility. AB - We report two empirical studies that investigated previously reported benefits of a high accuracy motivation, and thus a high threshold, for children's and adults' event recall and for their ability to resist false suggestions. In the studies, 6 , 7-, and 8-year-olds, as well as adults, were shown a brief video about an event and were later asked unbiased and misleading questions about it. In Study 1, participants were either (a) given the typical accuracy instructions (including the option to answer with "I don't know"), (b) reminded of the accuracy instructions during the interview, or (c) immediately given feedback and a token for every correct answer. The results showed that the reminders were ineffective in stimulating strategic control behavior in children, independent of age. In Study 2, the confounding effects of feedback and incentives were disentangled by contrasting (a) free report, (b) feedback only, (c) incentives only, and (d) feedback plus incentives. Analyses on recall performance and suggestibility revealed that both feedback and incentives are necessary to increase children's accurate memory reports. PMID- 15814095 TI - Updating in working memory: a comparison of good and poor comprehenders. AB - In this research, we examined the relation between reading comprehension and success in a working memory updating task. We tested the hypotheses that poor comprehenders' deficiencies are associated with a specific difficulty in the working memory updating process, particularly in controlling for information that is no longer relevant. In the first experiment, groups of poor and good comprehenders, ages 8-11 years, were administered a working memory updating task. In the second experiment a year later, a subgroup of participants involved in the first experiment was tested with a different updating task. In both experiments, poor comprehenders had less accurate recall performance and made more intrusion errors than did good comprehenders. Moreover, distinguishing intrusion errors on the basis of their permanence in memory, we found that poor comprehenders were more likely to intrude items that were maintained longer in memory than were good comprehenders. This type of error predicted reading comprehension abilities better than did working memory recall. This suggests that the relation between reading comprehension and working memory is mediated by the ability to control for irrelevant information. PMID- 15814096 TI - The development of interpretations for novel noun-noun conceptual combinations during the early school years. AB - This research investigates issues surrounding early school children's use of the similarity between head and modifier terms in deriving interpretations for novel noun-noun conceptual combinations. In these experiments, 6- and 9-year-olds and adults were asked to formulate interpretations of similar and dissimilar conceptual combinations. Both children and adults were sensitive to the similarity aspect of conceptual combinations, although the children had some difficulty with the property interpretations that high-similarity combinations require. Next, we examined 40 popular children's books for the presence of noun noun conceptual combinations. Adult participants provided interpretations for these combinations and rated the similarity of the head and modifier nouns. Results indicated that there were few high-similarity combinations and few combinations requiring property interpretations, suggesting that children have limited exposure to highly similar combinations and property interpretations. Further analysis of children's interpretations indicates that they may have difficulty in selecting and integrating properties of the modifier onto the head, a process required by property interpretations. PMID- 15814097 TI - The GABAA receptor agonist THIP alters the EEG in waking and sleep of mice. AB - THIP is a GABA(A) agonist with hypnotic properties consisting in reducing sleep latency and prolonging and consolidating sleep. THIP has been reported to increase EEG slow-wave activity (SWA; EEG power in the 0.75-4 Hz band) in non-REM (NREM) sleep in both rats and humans. We investigated the effects of THIP on sleep in C57BL/6 mice. EEG recordings were performed after 2, 4 and 6 mg/kg THIP and saline control. The results were compared with analyses of recordings obtained after 6 h of sleep deprivation (SD) in the same strain of mice. The two higher doses of THIP induced an abnormal EEG pattern both in waking and NREM sleep. The EEG was characterized by sporadic asymmetric high-voltage potentials recurring at a low-frequency (<1 Hz) on the background of a low-amplitude EEG pattern. In contrast, after SD the typical regular synchronous high amplitude delta waves predominated. THIP at 4 and 6 mg/kg led to a prominent enhancement of spectral power in the low-frequency range of the waking and sleep EEG which was much higher than the increase attained after 6 h SD. This effect was particularly prominent in the waking EEG. In NREM sleep the increase of spectral power after THIP reflected the frequency of recurrence of the high-voltage potentials, and was restricted to a narrower frequency band than after SD. The EEG changes after 2mg/kg differed little from saline control. Sleep latency was not affected by the two lower doses of THIP, and was prolonged after 6 mg/kg. REM sleep was suppressed after the two higher doses. In contrast to previous results reported in other species, THIP did not have a hypnotic action in mice. The changes induced by THIP in the waking and sleep EEG differed from those caused by enhanced physiological sleep pressure encountered after SD. Considering the abnormal EEG pattern and the similarity of the spectral changes in the sleep and waking EEG, THIP does not seem to exert a specific effect on mechanisms involved in sleep regulation. PMID- 15814099 TI - Opioids inhibit purinergic nociceptors in the sensory neurons and fibres of rat via a G protein-dependent mechanism. AB - We have found that opioid and P2X receptors are functionally coupled in the sensory nerve fibres and neurons of rat. When examined in the skin-nerve preparation, the ATP-evoked discharges of nerve fibres belonging to n. saphenous were inhibited by various opiates in a naloxone-dependent manner. The functional coupling between opioid and purinergic receptors was studied in the neuronal cell bodies isolated from dorsal root and nodose ganglia. Both fast (mediated by P2X(3) receptors) and slow (P2X(2/3) heteromeric receptors) responses of sensory neurons to ATP were inhibited by opioids. The inhibition of slow responses developed in a characteristic biphasic manner: an initial short phase of potentiation (lasting for 300-400 s) was followed by long-lasting inhibition of the response (for about 50% when saturated). Both phases of the response were initiated by the application of the highly selective ligand for mu-receptors, endomorphin 1 (30 nM). Intracellular GTPgammaS caused a partial inhibition of the ATP responses and opioids were not effective against the residual response. Intracellular GDP eliminated the effects of opioids, while pertussis toxin (PTX) abolished only the inhibition phase. Thus, P2X receptors in the sensory neurons are affected by opioids via multiple G protein-dependent pathways. PMID- 15814098 TI - Reversal of caffeine-induced anxiety by neurosteroid 3-alpha-hydroxy-5-alpha pregnane-20-one in rats. AB - Caffeine has been shown to increase brain and plasma content of neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP) that allosterically modulates GABA(A) receptors. The present study evaluated the role of neurosteroid 3alpha,5alpha-THP in the caffeine-induced anxiogenic-like effect using the elevated plus-maze (EPM) test in rats. Acute administration of caffeine (50 or 100mg/kg, i.p.) produced anxiogenic-like activity that was reversed by pretreatment with the neurosteroid 3alpha,5alpha-THP or progesterone, the GABA(A) agonist muscimol, or the benzodiazepine receptor agonist diazepam. On the contrary, caffeine produced higher anxiety in animals previously treated with the GABA(A) receptor antagonist, bicuculline or either of the various neurosteroid biosynthesis enzyme inhibitors viz. trilostane, finasteride or indomethacin. Furthermore, pretreatment with DHEAS, a neurosteroid that negatively modulates GABA(A) receptors also enhanced the caffeine-induced anxiety. Moreover, adrenalectomy potentiated the anxiogenic-like response of caffeine indicating the contributory role of peripheral steroidogenesis. Thus, it is speculated that neurosteroid 3alpha,5alpha-THP through positive modulation of GABA(A) receptor activity may serve as a counter-regulatory mechanism against caffeine-induced anxiety. PMID- 15814100 TI - Alterations in brain Protein Kinase A activity and reversal of morphine tolerance by two fragments of native Protein Kinase A inhibitor peptide (PKI). AB - Two peptide fragments of native Protein Kinase A inhibitor (PKI), PKI-(6-22) amide and PKI-(Myr-14-22)-amide, significantly reversed low-level morphine antinociceptive tolerance in mice. The inhibition of Protein Kinase A (PKA) activity by both peptide fragments was then measured in specific brain regions (thalamus, periaqueductal gray (PAG), and medulla) and in lumbar spinal cord (LSC), which in previous studies have been shown to play a role in morphine induced analgesia. In drug naive animals, cytosolic PKA activity was greater than particulate PKA activity in each region, while cytosolic and particulate PKA activities were greater in thalamus and PAG compared to medulla and LSC. The addition of both peptides to homogenates from each region completely abolished cytosolic and particulate PKA activities in vitro. Following injection into the lateral ventricle of the brain of drug naive mice and morphine-tolerant mice, both peptides inhibited PKA activity in the cytosolic, but not the particulate fraction of LSC. In addition, cytosolic and particulate PKA activities were inhibited by both peptides in thalamus. These results demonstrate that the inhibition of PKA reverses morphine tolerance. Moreover, the inhibition of PKA activity in specific brain regions and LSC from morphine-tolerant mice by PKI analogs administered i.c.v. is evidence that PKA plays a role in morphine tolerance. PMID- 15814101 TI - Pharmacological and pharmacokinetic characterization of the cannabinoid receptor 2 agonist, GW405833, utilizing rodent models of acute and chronic pain, anxiety, ataxia and catalepsy. AB - To date, two cannabinoid receptors have been identified, CB1 and CB2. Activation of these receptors with non-selective cannabinoid receptor agonists reduces pain sensitivity in animals and humans. However, activation of CB1 receptors is also associated with central side effects, including ataxia and catalepsy. More recently, a role for selective CB2 agonists in pain modification has been demonstrated. GW405833, a selective CB2 agonist, was recently reported to partially reverse the inflammation and hyperalgesia in a rat model of acute inflammation. In the current report, we extend the characterization and therapeutic potential of this compound. For the first time, we show that GW405833 selectively binds both rat and human CB2 receptors with high affinity, where it acts as a partial agonist (approximately 50% reduction of forskolin-mediated cAMP production compared to the full cannabinoid agonist, CP55,940). We also report for the first time that intraperitoneal administration of GW405833 (0.3-100 mg/kg) to rats shows linear, dose-dependent increases in plasma levels and substantial penetration into the central nervous system. In addition, GW405833 (up to 30 mg/kg) elicits potent and efficacious antihyperalgesic effects in rodent models of neuropathic, incisional and chronic inflammatory pain, the first description of this compound in these models. In contrast, analgesia, sedation and catalepsy were not observed in this dose range, but were apparent at 100 mg/kg. Additionally, GW405833 was not antihyperalgesic against chronic inflammatory pain in CB2 knockout mice. These data support the tenet that selective CB2 receptor agonists have the potential to treat pain without eliciting the centrally-mediated side effects associated with non-selective cannabinoid agonists, and highlight the utility of GW405833 for the investigation of CB2 physiology. PMID- 15814102 TI - Effect of acute and chronic psychostimulant drugs on redox status, AP-1 activation and pro-enkephalin mRNA in the human astrocyte-like U373 MG cells. AB - In order to approach the astroglial implication of addictive and neurotoxic processes associated with psychostimulant drug abuse, the effects of amphetamine or cocaine (1-100 microM) on redox status, AP-1 transcription factor and pro enkephalin, an AP-1 target gene, were investigated in the human astrocyte-like U373 MG cells. We demonstrated an early increase in the generation of radical oxygen species and in the formation of 4-hydroxynonenal-adducts reflecting the pro-oxidant action of both substances. After 1 h or 96 h of treatment, Fos and Jun protein levels were altered and the DNA-binding activity of AP-1 was increased in response to both substances. Using supershift experiments, we observed that the composition of AP-1 dimer differed according to the substance and the duration of treatment. FRA-2 protein represented the main component of the chronic amphetamine- or cocaine-activated complexes, which suggests its relevance in the long-term effects of psychostimulant drugs. Concomitantly, the pro-enkephalin gene was differently regulated by either 6 h or 96 h of treatment. Because astrocytes interact extensively with the neurons in the brain, our data led us to conclude that oxidation-regulated AP-1 target genes may represent one of the molecular mechanisms underlying neuronal adaptation associated with psychostimulant dependence. PMID- 15814103 TI - Gene - environment interactions determine the individual variability in cocaine self-administration. AB - Research into factors that determine the propensity to self-administer cocaine has shown that stressors can determine the amount of cocaine self-administered as well as the rate of acquisition. However, the interaction between the genetic make-up of the animal and stress is unknown. This study investigated this interaction by using the genetic animal model consisting of apomorphine susceptible (APO-SUS) and unsusceptible (APO-UNSUS) rats. Animals were allowed to self-administer 0.25 mg/kg cocaine under stressful and habituated conditions. This study revealed that the amount of cocaine consumed was highly dependent on the genetic make-up of the animal as well as the amount of stress during self administration. Under habituated circumstances the APO-UNSUS rats took far more cocaine than the APO-SUS rats. Under stressful circumstances, however, the APO SUS rats took far more cocaine than the APO-UNSUS rats. This difference in the amount consumed by APO-SUS and APO-UNSUS rats is likely to be due to the specific neurobiological features of their dopaminergic and, possibly, noradrenergic system as well as the reactivity of their HPA-axis. It is suggested that the amount of a drug consumed and, accordingly, its addictive potential and 'drug vulnerability' are determined by the interaction between the genetic make-up of the animals and stress, and not by either component alone. PMID- 15814104 TI - The subunit composition and pharmacology of alpha-Conotoxin MII-binding nicotinic acetylcholine receptors studied by a novel membrane-binding assay. AB - The subunit composition and pharmacology of alpha-Conotoxin MII-binding (alpha CtxMII) nicotinic acetylcholine receptors (nAChR) was studied by an improved [(125)I]-alpha-CtxMII membrane binding method. This binding method facilitates pharmacological studies that have been difficult to accomplish with [(125)I] alpha-CtxMII autoradiography or alpha-CtxMII inhibition of [(125)I]-epibatidine binding. Binding densities and K(d)-values obtained by this [(125)I]-alpha-CtxMII membrane binding were similar to the values obtained by autoradiography or alpha CtxMII inhibition of [(125)I]-epibatidine binding, verifying that each of these approaches measures the same nAChR population. Binding results with nAChR subunit null mutant mice confirm and extend observations from earlier studies: [(125)I] alpha-CtxMII binding measures two sets of alpha6beta2* nAChR (alpha4alpha6beta2beta3 or alpha6beta2beta3). Most nicotinic agonists and antagonists show monophasic inhibition of [(125)I]-alpha-CtxMII binding, indicating that alpha4alpha6beta2beta3 and alpha6beta2beta3 have similar binding properties. Comparison of the binding and activation profiles of alpha6beta2* nAChR to those of other nAChR subtypes (alpha4beta2* and beta4*) indicates that these receptors have distinctly different pharmacology indicating that it may be possible to target alpha6beta2* nAChR selectively to develop compounds that might be therapeutically useful. PMID- 15814105 TI - A comparison of neurokinin 1 receptor knock-out (NK1-/-) and wildtype mice: exploratory behaviour and extracellular noradrenaline concentration in the cerebral cortex of anaesthetised subjects. AB - In behavioural screens, mice lacking functional NK1 receptors (NK1-/-) resemble wildtypes (NK1+/+) that have been given an antianxiety/antidepressant drug. Most, if not all, antidepressants increase noradrenergic transmission in the brain. Here, we have used in vivo microdialysis to compare the concentrations of extracellular noradrenaline ('efflux') in the cerebral cortex of anaesthetised NK1-/- and NK1+/+ mice. The effects of systemic administration of the antidepressant, desipramine, with and without local infusion of the alpha(2) adrenoceptor antagonist, RX821002, were also evaluated. Finally, we compared the effects of desipramine on behaviour of NK1+/+ and NK1-/- mice in an activity chamber and in a light/dark exploration box. Basal noradrenaline efflux was increased 2 to 4-fold in NK1-/- mice compared with NK1+/+ mice but there was no difference in the effects of desipramine. RX821002 increased noradrenaline efflux in all vehicle-injected mice but, in desipramine-pretreated mice, noradrenaline efflux was increased in NK1+/+ mice, only. All behaviours in the light/dark exploration box differed in the two genotypes. Furthermore, with the exception of 'grooming', the effects of desipramine on behaviour of NK1-/- mice could be explained by the effects of this antidepressant on locomotor activity. Finally, alpha(2)-adrenoceptors are possibly desensitised in NK1-/- mice. We have yet to establish whether this is a cause or a consequence of the increased noradrenaline efflux. PMID- 15814106 TI - Iron protects astrocytes from 6-hydroxydopamine toxicity. AB - The role of iron in 6-hydroxydopamine (6-OHDA) toxicity towards astrocytes was investigated in vitro using rat primary astrocytes, rat astrocytoma cell line C6, and human astrocytoma cell line U251. The assessment of mitochondrial respiration or lactate dehydrogenase release has shown a dose-dependent decrease in the viability of astrocytes treated with 6-OHDA, which coincided with DNA fragmentation and the changes in cellular morphology. This was a consequence of the oxidative stress mediated by 6-OHDA autoxidation products hydrogen peroxide, superoxide anion, and hydroxyl radical. Both FeSO(4) and FeCl(3) markedly alleviated detrimental effects of 6-OHDA treatment, while MgSO(4) was without effect. The protective action of iron was neutralized by a membrane-permeable iron chelator o-phenanthroline, which also augmented astrocyte killing in the absence of exogenous iron. The mechanisms responsible for iron-mediated protection of astrocytes did not involve interference with either 6-OHDA autoxidation, hydrogen peroxide toxicity, or 6-OHDA-induced activation of extracellular signal-regulated kinase. Finally, the addition of iron potentiated and its chelation blocked 6-OHDA toxicity towards neuronal PC12 cells, suggesting the opposite roles for this transition metal in regulating the survival of astrocytes and dopaminergic neurons. PMID- 15814107 TI - CCK-8 prevents the development of kindling and regulates the GABA and NPY expression in the hippocampus of pentylenetetrazole (PTZ)-treated adult rats. AB - Neuronal loss and irreversible brain damage often cause the worsening of symptoms and the decreased efficacy of pharmacological treatment occurring in epileptic patients and animal models of kindling. Recently we reported that the neurotransmitter/neuromodulatory peptide Cholecystokinin-8 (CCK-8) is able to induce the structural and functional neuronal recovery of chemical- and surgical induced lesions when i.p. injected in rodents. The present study therefore, was aimed at verifying the hypothesis that treatment with a CCK-8 dose having a neuroprotective action might affect brain alterations and the development of kindling in adult rats receiving the convulsant agent pentylenetetrazole (PTZ). Compared to rats receiving Saline prior to PTZ, which manifested clonic-tonic seizures (Class 5 behavioural change scale) after three weeks of treatment, rats pre-treated with CCK-8 showed an improvement of behavioural score exhibiting myoclonus and occasionally tonic seizures (Class 3/4). This decreased susceptibility to develop convulsions was associated with the recovery of PTZ induced reduction of ChAT levels in forebrain and GABA/GAD expression in the hippocampus. Furthermore, NPY immunoreactivity distribution and NPY mRNA levels were also increased in the hippocampus of rats receiving CCK-8 injection before each PTZ treatment. These data indicate that CCK-8 possesses the ability to prevent and/or suppress the convulsant effects of PTZ by stimulating the synthesis of neurotransmitters/peptides involved in the inhibition of hippocampal hyper-excitability. Our findings suggest that CCK-8 may have anticonvulsant and neuroprotective properties that merit further investigation. PMID- 15814108 TI - Protective effects of ginseng saponins on 3-nitropropionic acid-induced striatal degeneration in rats. AB - The precise cause of neuronal cell death in Huntington's disease (HD) is not known. Systemic administration of 3-nitropropionic acid (3-NP), an irreversible succinate dehydrogenase inhibitor, not only induces a cellular ATP depletions but also causes a selective striatal degeneration similar to that seen in HD. Recent accumulating reports have shown that ginseng saponins (GTS), the major active ingredients of Panax ginseng, have protective effects against neurotoxin insults. In the present study, we examined in vitro and in vivo effects of GTS on striatal neurotoxicity induced by repeated treatment of 3-NP in rats. Here, we report that systemic administration of GTS produced significant protections against systemic 3-NP- and intrastriatal malonate-induced lesions in rat striatum with dose dependent manner. GTS also improved significantly 3-NP-caused behavioral impairment and extended survival. However, GTS itself had no effect on 3-NP induced inhibition of succinate dehydrogenase activity. To explain the mechanisms underlying in vivo protective effects of GTS against 3-NP-induced striatal degeneration, we examined in vitro effect of GTS against 3-NP-caused cytotoxicity using cultured rat striatal neurons. We found that GTS inhibited 3-NP-induced intracellular Ca(2+) elevations. GTS restored 3-NP-caused mitochondrial transmembrane potential reduction in cultured rat striatal neurons. GTS also prevented 3-NP-induced striatal neuronal cell deaths with dose-dependent manner. The EC(50) was 12.6 +/- 0. 7microg/ml. These results suggest that in vivo protective effects of GTS against 3-NP-induced rat striatal degeneration might be achieved via in vitro inhibition of 3-NP-induced intracellular Ca(2+) elevations and cytotoxicity of striatal neurons. PMID- 15814109 TI - Signal transduction underlying carbachol-induced PGE2 generation and cox-1 mRNA expression of rat brain. AB - In this paper we have determined the different signal pathways involved in M(1) and M(3) muscarinic acetylcholine receptor (mAChR) dependent stimulation of cyclo oxygenase 1 (cox-1) mRNA gene expression and PGE(2) production on rat cerebral frontal cortex. Carbachol stimulation of M(1) and M(3) mAChR exerts an increase in cox-1 mRNA gene expression without affecting cox-2 mRNA expression and increased PGE(2) generation. Besides, increased phosphoinositide (PI) turnover and stimulation of nitric oxide synthase (NOS) and cyclic GMP (cGMP) production. Inhibitors of phospholipase A(2) (PLA(2)), COX and phospholipase C (PLC), calcium/calmodulin (CaM), NOS and soluble guanylate cyclase prevent the carbachol effect. These results suggest that carbachol-activation of M(1) and M(3) mAChR increased PGE(2) release associated with an increased expression of cox-1 and NO cGMP production. The mechanism appears to occur directly to PLC stimulation and indirectly to PLA(2) activation. These results may contribute to understand the effects and side effect of non-steroidal anti-inflammatory drugs in patients with cerebral degenerative diseases. PMID- 15814112 TI - Cutaneous lasers. AB - Over the past few decades, lasers have gained increasing use in medicine. Clinical indications for the use of lasers in plastic surgery include facial rejuvenation, tattoo removal, hair removal, and improvement in various congenital and acquired skin abnormalities. With the advent of new lasers, patients of all ethnic backgrounds can be treated effectively. PMID- 15814110 TI - Differential expression of P2X receptors on neurons from different parasympathetic ganglia. AB - Whole-cell patch clamp recording and immunohistochemistry were used to investigate the expression of P2X receptors on rat parasympathetic ganglion neurons of the otic, sphenopalatine, submandibular, intracardiac and paratracheal ganglia. Neurons from all five ganglia responded to ATP with a rapidly activating, sustained inward current. Neurons of intracardiac and paratracheal ganglia were insensitive to alphabeta-meATP, while all neurons in the otic and some neurons of sphenopalatine and submandibular ganglia responded. Lowering pH potentiated ATP responses in neurons from all five ganglia. Co-application of Zn(2+) potentiated ATP responses in intracardiac, paratracheal and submandibular ganglion neurons. Immunohistochemistry revealed strong and specific staining for the P2X(2) subunit in all five ganglia and strong P2X(3) staining in otic, sphenopalatine and submandibular ganglia. In conclusion, there is heterogeneity in P2X receptor expression in different parasympathetic ganglia of the rat, but the predominant receptor subtypes involved appear to be homomeric P2X(2) and heteromeric P2X(2/3). PMID- 15814113 TI - Injectable skin fillers. AB - The list of injectable skin fillers available for soft tissue augmentation is constantly growing, giving aesthetic surgeons more options in the treatment of scars, lines, and wrinkles. Each type of filler is distinct with regard to intended use, placement technique, duration in tissue, ease of use, and cost. Physicians who are well acquainted with the strengths and weaknesses of each class of filler will be best equipped to use this rapidly expanding class of agents in serving their patients. The authors review the most commonly used injectable skin fillers and discuss their differences in terms of composition, source (eg, human, animal), intended use, placement technique, tissue duration (nonpermanent, semipermanent, or permanent), side effects, and specific advantages and disadvantages. PMID- 15814114 TI - Hair restoration. AB - The introduction of micrografts (1-2 hair follicular unit grafts) and minigrafts (3-4 hair follicular unit grafts) has made a most significant advancement in the care of male pattern baldness and female androgenic alopecia. Finally, natural and aesthetically pleasing results are possible. Additionally, many other applications in the reconstruction of facial hair and scalp have been found, some of these include: restoration of hair loss due to (iatrogenic) post-surgical causes, ie, after facial rejuvenation procedures or procedures involving incisions on hair bearing facial skin or scalp; scalp and facial hair due to burns or to traumatic injuries; post-oncologic resections; hair loss due to congenital reasons such as in cases of complete bilaterial cleft lips (no mustache hair in the prolabium). PMID- 15814115 TI - Cutaneous vascular lesions. AB - Vascular anomalies are lesions seen in all surgical disciplines, particularly in pediatric patients. Specialization in vascular anomalies involves a team effort, with the team consisting of plastic surgeons, general surgeons, neurosurgeons, pediatricians, interventional radiologists, dermatologists, ophthalmologists, otolaryngologists, hematologists, and pathologists. Inconsistent nomenclature in the literature has historically resulted in confusion about classification, diagnosis, and treatment. A biologic classification system has emerged, based on clinical observations, natural history, and cellular features, which separates vascular anomalies into two broad categories: vascular tumors and vascular malformations. For many vascular anomalies, photodocumentation, psychosocial support, and communication are important throughout the treatment course. PMID- 15814116 TI - Acute burn injury. AB - Almost 2 million people in the United States suffer from burns annually. A small percentage of these injuries are fatal, but all require some degree of medical attention. Burn injury is associated with anatomic, physiologic, endocrinologic, and immunologic alterations. These problems need to be identified and treated properly to prevent or minimize the extent of the damage. In recent years, advances in burn treatment have reduced morbidity and mortality and improved the quality of life for burn survivors. These advances have been made in the treatment of the acute injury, the quality of the initial resuscitation, the effectiveness of infection control, and the surgical decision making for improved short- and long-term outcomes. PMID- 15814117 TI - Acute wounds. AB - The most important factors in the management of acute wounds are the history and physical examination. The goals of wound care are fivefold: avoid further tissue damage, achieve wound closure as rapidly as possible, restore function to the injured tissue, facilitate the patient's expedient return to normal daily activities, and restore the patient's quality of life. The treating physician must have a good understanding of the wound healing mechanism. One must rule out all associated occult injuries that may be life threatening. Proper wound assessment and management with minimal discomfort to the patient are crucial. The primary goal is to facilitate the healing process to achieve a cosmetically pleasing and functional result. PMID- 15814118 TI - Chronic wounds. AB - Chronic wounds are a challenge to treat for the clinician. We present a current overview of intrinsic and extrinsic factors in the development chronic nonhealing wounds. Solutions to some of these difficult problems are presented. PMID- 15814119 TI - Premalignant and early squamous cell carcinoma. AB - Actinic keratosis (AK) is a common sun-induced precancerous neoplasm confined to the epidermis. The AK is the initial manifestation of a continuum of clinical and histologic abnormalities that progresses to invasive squamous cell carcinoma (SCC). Bowen's disease, also known as squamous cell carcinoma in situ, represents early SCC confined to the epidermis. More than half of all SCCs contain p53 tumor suppressor gene mutations. Like SCCs, the vast majority of AKs and Bowen's disease lesions are asymptomatic. Each AK and suspicious lesion should be treated before it progresses to invasive SCC. Destructive modalities, such as cryosurgery using liquid nitrogen and electrodesiccation and curettage, usually performed by a dermatologist, are the mainstays of therapy. PMID- 15814120 TI - Nonmelanoma skin cancer. AB - Each year, there are as many cases of nonmelanoma skin cancer as all other cancers combined. Although there is relatively low attributable mortality, the morbidity and expense of treatment is significant. Unlike many other malignancies, host and environmental factors relevant to the pathophysiology have been clearly demonstrated. Surgical ablation remains the mainstay of treatment. PMID- 15814121 TI - Melanoma. AB - The surgical management of melanoma continues to evolve. A large body of information serves as a foundation for the oncologic principles, surgical excisions, and reconstructive methodologies that are currently in use. This article serves as a guide for the physician considering surgical management of the melanoma patient. PMID- 15814122 TI - Skin flaps. AB - Open wounds, particularly around the face, often require complicated techniques for optimal closure. The approach to the closure of the complicated wound depends largely on the nature of the wound, including the location and size of the defect, the functional outcome after closure, the medical comorbidities of the patient, neighboring structures, and whether the defect is secondary to a malignancy or trauma. The goals of wound management are optimal aesthetic outcome, preservation of function, and patient satisfaction. The authors briefly review basic skin closure options and discuss use of skin flaps, particularly of the head and neck region. PMID- 15814123 TI - Reconstruction of complex facial defects after radical resection of advanced skin cancers. AB - Management of patients with extensive facial skin cancers is a challenge. We present a series of nine patients with extensive facial skin cancers and our experience with managing these patients. PMID- 15814124 TI - Immobilization of hemoglobin at the galleries of layered niobate HCa2Nb3O10. AB - Hemoglobin (Hb) was intercalated at the galleries of layered niobate HCa(2)Nb(3)O(10) (HCNO). Two different kinds of layered phases of Hb-CNO composites Hb-CNO-1 and Hb-CNO-2 were obtained with the interlayer distances of 7.2 and 10.3nm in correspondence with the monolayer and bilayer arrangements of proteins between the niobate layers, respectively, based on the powder XRD pattern, HRTEM, UV-vis spectra and CHN analyses. FTIR spectra of Hb-CNO composites show that amide I and amide II bands were actually the same as those of the native Hb, which indicates that there is almost no structural change after immobilization. Michaelis-Menten model methods were used to study the peroxidatic activity of the reaction of 2-methoxyphenol and H(2)O(2) for the entrapped Hb in the galleries of HCNO. Compared to that of free Hb, the kinetic parameters of Hb CNO k(cat), K(M) and k(cat)/K(M) were affected by the immobilization process. The immobilized Hb showed a higher relative activity than that of free Hb after incubated in phosphate buffer (pH = 7) at 80 degrees C for a period of time. The environments between the layers of HCNO are hydrophilic which will bind water tightly and help to stabilize the 'essential water' layer around the protein. So, immobilization of Hb between the layers of HCNO enhanced the activity of Hb in water-DMSO mixture. PMID- 15814125 TI - Preparation and characterization of porous beta-tricalcium phosphate/collagen composites with an integrated structure. AB - Porous beta-tricalcium phosphate (TCP)/collagen composites with different beta TCP/collagen weight ratio were prepared. The influences of the preparation conditions on the microstructure of porous composite and the joint status of beta TCP particles with collagen fibrils were characterized by X-ray diffractometer, scanning electron microscopy and transmission electron microscopy. The results showed: (1) an acid treatment could effectively disassemble collagen fibrils; (2) in the resulting porous composites, beta-TCP particles homogenously existed on the skeleton of the collagen fibril network and bonded tightly to both the fibrils and themselves. The tight bonding formation could be due to the reaction between Ca ions in the particles and carboxyl groups in collagen polypeptide chains and due to the reprecipitation of partially dissolved beta-TCP during synthesis. The tight bonding between beta-TCP particles and collagen fibrils in the composites demonstrated an integrated structure, which was reproducible when beta-TCP/collagen ratio ranged from 2 to 4. Such integrated structure would make significant contributions in reliably tailoring properties of the porous composites by varying beta-TCP content. In addition, the porous composites had large porosity (approximately 95%) and appropriate pore size (approximately 100 microm), showed no negative impact in cytotoxicity assay and complete bone tissue regeneration after 12 weeks in animal test. PMID- 15814126 TI - Platelet interactions with calcium-phosphate-coated surfaces. AB - Many studies have shown that calcium-phosphate (CaP)-coated endosseous implants exhibit more peri-implant bone formation and bone contact at early healing times than uncoated implants. Since the rate of healing is influenced by blood/implant interactions and possibly the degree of blood platelet activation, the aim of this study was to determine whether the topography, microtopography, or the presence of calcium (Ca) and phosphate (PO(4)) ions in the implant surface plays a predominant role in platelet activation. We define the threshold between topography and microtopography as the limit of the scale range of platelets themselves; thus, a microtopographic surface is defined by one which exhibits features 3mum. With the help of four international collaborating laboratories, we prepared 11 titanium and CaP-modified titanium surfaces each with different (micro)topographies and interrogated these surfaces with both platelet adhesion (lactate dehydrogenase activity) and platelet activation (microparticle formation and P-selectin expression) assays. Our results show that: calcium (Ca)- and phosphate (PO(4))-containing surfaces of increasing surface microtopographical complexity exhibit increasing platelet activation; surfaces with similar surface microtopographies show similar levels of platelet activation regardless of the presence of Ca and PO(4) in the surface; and that surface microtopography is responsible for platelet activation rather than the presence of Ca and PO(4) in the surface. PMID- 15814127 TI - The quantification of cellular viability and inflammatory response to stainless steel alloys. AB - The biocompatibility of metallic alloys is critical to the success of many orthopedic therapies. Corrosion resistance and the immune response of the body to wear debris products ultimately determine the performance of these devices. The establishment of quantitative tests of biocompatibility is an important issue for biomaterials development. We have developed an in vitro model to measure the pro inflammatory cytokine production and in this study investigated the cellular responses induced by nitrogenated and 316L stainless steel alloys in both particulate and solid form. We utilized a murine macrophage cell line, RAW 264.7, to characterize and compare the mRNA profiles of TNF-alpha and IL-1beta in these cells using real time-polymerase chain reaction (RT-PCR). Fluorescence microscopy and flow cytometry were used to probe the viability of the population and to examine the apoptotic pathway. The goals of this work were to develop improved measurement methods for the quantification of cellular inflammatory responses to biomaterials and to obtain data that leads to an enhanced understanding of the ways in which the body responds to biomaterials. Using these techniques, we observed evidence for an association between the upregulation of IL-1beta and reversible apoptosis, and the upregulation of TNF-alpha and irreversible apoptosis. PMID- 15814128 TI - In vitro bioactivity of MOEP grafted ePTFE membranes for craniofacial applications. AB - The bioactivity of three methacryloyloxyethyl phosphate (MOEP) grafted expanded polytetrafluoroethylene (ePTFE) membranes with varying surface coverage as well as unmodified ePTFE was investigated through a series of in vitro tests: calcium phosphate (CaP) growth in simulated body fluid (SBF), serum protein adsorption, and a morphology and attachment study of human osteoblast-like SaOS-2 cells. The graft copolymers were prepared by means of gamma irradiation induced grafting and displayed various surface morphologies and wettabilities depending on the grafting conditions used. Unmodified ePTFE did not induce nucleation of CaP minerals, whereas all the grafted membranes revealed the growth of CaP minerals after 7 days immersion in SBF. The sample with lowest surface grafting yield (24% coverage), a smooth graft morphology and relatively high hydrophobicity (theta(adv) = 120 degrees, theta(rec) = 80 degrees) showed carbonated hydroxyapatite growth covering the surface. On the other hand, the samples with high surface grafting yield (76% and 100%), a globular graft morphology and hydrophilic surfaces (theta(adv) = 60 degrees and 80 degrees, theta(rec) = 25 degrees and 15 degrees, respectively) exhibited irregular growth of non-apatitic CaP minerals. Irreversibly adsorbed protein measured after a 1h immersion in serum solution was quantified by the amount of nitrogen on the surface using XPS, as well as by weight increase. All grafted membranes adsorbed 3-6 times more protein than the unmodified membrane. The sample with the highest surface coverage adsorbed the most protein. Osteoblast-like SaOS-2 cells cultured for 3 h revealed significantly higher levels of cell attachment on all grafted membranes compared to unmodified ePTFE. Although the morphology of the cells was heterogeneous, in general, the higher grafted surfaces showed a much better cell morphology than both the low surface-grafted and the control unmodified sample. The suite of in vitro tests confirms that a judicious choice of grafted monomer such as the phosphate-containing methacrylate monomer (MOEP) significantly improves the bioactivity of ePTFE in vitro. PMID- 15814129 TI - The inflammatory potential of biphasic calcium phosphate granules in osteoblast/macrophage co-culture. AB - Some biological properties of a range of biphasic calcium phosphate (BCP) granules were quantified using a co-culture model of primary unstimulated human osteoblasts and macrophages. The BCP granules were classified in terms of diameter and the ratio of tricalcium phosphate (TCP): hydroxyapatite (HA). All granules were 50% porous and sterilised by gamma-irradiation. Primary unstimulated human osteoblasts and macrophages were cultured in contact with the range of BCP granules for 1, 7 and 14 days. The response of the cells was determined and quantified by the assessment of viable cell adhesion to the substrate, using lactate dehydrogenase assay, and the production and release of the cytokines; interleukin 1beta, (IL-1beta), tumour necrosis factor alpha (TNF alpha) and prostaglandin E2 (PGE2). Throughout the test period viable cell adhesion on all BCP granules was significantly lower than the tissue culture polystyrene control. Higher content TCP materials, (80% and 100% TCP) did not support viable cell adhesion after 1 day, lower content TCP materials, (20% and 50% TCP) granules did support viable cell adhesion throughout the time period. The percentage content of TCP was a more significant factor than granule size within the test conditions at all time points. PMID- 15814130 TI - The effect of antioxidants on oxidative DNA damage induced by visible-light irradiated camphorquinone/N,N-dimethyl-p-toluidine. AB - Previous investigations have found that visible-light (VL)-irradiated camphorquinone (CQ), in the presence of a tertiary amine (e.g., N,N-dimethyl-p toluidine, DMT), generates reactive oxygen species and causes oxidative DNA damage in vitro. In this study, oxidative DNA damage produced by VL-irradiated CQ/DMT, in the presence and absence of antioxidants (glutathione, N-acetyl-L cysteine (NAC), mannitol, vitamin C, and vitamin E), was measured by the conversion of PhiX-174 RF I supercoiled (SC) double-stranded plasmid DNA into open and linear forms. VL-irradiated CQ/DMT, lacking antioxidant, damaged 99.4 +/ 1% of the PhiX-174 RF I SC double-stranded plasmid DNA. Our results revealed that glutathione (10.0, 5.0, 2.5, 1.0, and 0.5 mm) and NAC (10.0, 5.0, and 2.5 mm) significantly (p < 0.02) reduced oxidative DNA damage produced by VL irradiated CQ/DMT. Vitamin E, vitamin C, and mannitol were ineffective at reducing oxidative DNA damage produced by VL-irradiated CQ/DMT. Furthermore, vitamin E (10.0 and 5.0 mm) and vitamin C (10.0, 5.0, 2.5, 1.0, 0.5 mm) treatment significantly (p < 0.02) enhanced VL-irradiated CQ/DMT-induced oxidative DNA damage and caused significant (p < 0.001) DNA damage following VL-irradiation in the absence of CQ/DMT. As a result, future studies should evaluate whether glutathione and NAC effectively reduce or prevent oxidative damage induced by VL irradiated CQ/DMT in vivo. PMID- 15814131 TI - Electrospun fine-textured scaffolds for heart tissue constructs. AB - The structural and functional effects of fine-textured matrices with sub-micron features on the growth of cardiac myocytes were examined. Electrospinning was used to fabricate biodegradable non-woven poly(lactide)- and poly(glycolide) based (PLGA) scaffolds for cardiac tissue engineering applications. Post processing was applied to achieve macro-scale fiber orientation (anisotropy). In vitro studies confirmed a dose-response effect of the poly(glycolide) concentration on the degradation rate and the pH value changes. Different formulations were examined to assess scaffold effects on cell attachment, structure and function. Primary cardiomyocytes (CMs) were cultured on the electrospun scaffolds to form tissue-like constructs. Scanning electron microscopy (SEM) revealed that the fine fiber architecture of the non-woven matrix allowed the cardiomyocytes to make extensive use of provided external cues for isotropic or anisotropic growth, and to some extent to crawl inside and pull on fibers. Structural analysis by confocal microscopy indicated that cardiomyocytes had a preference for relatively hydrophobic surfaces. CMs on electrospun poly(L-lactide) (PLLA) scaffolds developed mature contractile machinery (sarcomeres). Functionality (excitability) of the engineered constructs was confirmed by optical imaging of electrical activity using voltage-sensitive dyes. We conclude that engineered cardiac tissue structure and function can be modulated by the chemistry and geometry of the provided nano- and micro-textured surfaces. Electrospinning is a versatile manufacturing technique for design of biomaterials with potentially reorganizable architecture for cell and tissue growth. PMID- 15814132 TI - Chitosan-RGDSGGC conjugate as a scaffold material for musculoskeletal tissue engineering. AB - In the present study, we have developed a novel and versatile method for the preparation of chitosan-peptide complex based on the selective reaction of chitosan with 2-iminothiolane. The new type of SH-chitosan derivative showed an excellent solubility to aqueous solution even in the alkaline conditions. This characteristic greatly facilitated further modification study of chitosan with a variety of bioactive substances. A synthetic peptide, RGDSGGC containing RGDS moiety that is known as one of the most important cell adhesive peptides, was readily coupled by disulfide bonds formation with sulfhydryl groups of SH chitosan in the presence of dimethyl sulfoxide. Next, the effect of the introduction of RGDSGGC moiety to chitosan on cell adhesion and proliferation activity of chondrocytes and fibroblasts were evaluated. As a result, it was suggested that this polysaccharide-peptide conjugate exhibited excellent capacities for both cell adhesion and cell proliferation of chondrocytes and fibroblasts. Considering the growing importance of the biocompatible scaffolds in the recent tailored tissue engineering technique, these results indicate that the present strategy of 2-iminothiolane-based conjugation of polysaccharides with biologically active peptides will become a key and potential technology to develop desirable scaffold materials for the tissue regenerations. PMID- 15814133 TI - The influence of GFP-actin expression on the adhesion dynamics of HepG2 cells on a model extracellular matrix. AB - Integrins belong to a family of important cell surface receptors which mediate the adhesion of most anchorage-dependent cells to nature extracellular matrix (ECM) and biomaterials. It is known that the binding of integrin with ECM proteins triggers mechanochemical responses of cytoskeleton. To date, the intricate interplay between integrin-ECM interaction and cytoskeleton dynamics leading to the regulation of cell morphogenesis on biomaterials remains largely unknown. In this study, green fluorescence protein (GFP)-actins were expressed in HepG2 cells for the temporal visualization of cytoskeletal structure of adherent cells on naturally derived materials. By combining confocal reflectance contrast microscopy and fluorescence microscopy, the adhesion contact dynamics, cytoskeleton remodeling and two-dimensional spreading of intact and GFP-actin expressing HepG2 cells on collagen and fibronectin-coated substrates are simultaneously probed during the initial cell seeding. First of all, our results show that the evolution of adhesion contact of HepG2 cells upon integrin-collagen or integrin-fibronectin interaction is impaired by GFP-actin expression. Also, the initial rate of cell deformation is reduced by 70% and 43% on fibronectin and collagen, respectively, upon GFP-actin expression. Interestingly, the steady state adhesion energy of HepG2 cells remains unchanged and increases on fibronectin- and collagen-coated substrate, respectively, upon GFP-actin expression. Our highly integrated biophysical approach demonstrates that GFP actins diffusively concentrate in the cytoplasmic cortex during initial cell seeding while adhesion contact evolves and cell spreads. Kinetics analysis on the adhesion contact formation demonstrates the intricate interplay between cytoskeleton property and ECM proteins in cell adhesion. PMID- 15814134 TI - Behaviour of human endothelial cells on surface modified NiTi alloy. AB - Intravascular stents are being designed which utilise the shape memory properties of NiTi alloy. Despite the clinical advantages afforded by these stents their application has been limited by concerns about the large nickel ion content of the alloy. In this study, the surface chemistry of NiTi alloy was modified by mechanical polishing and oxidising heat treatments and subsequently characterised using X-ray photon spectroscopy (XPS). The effect of these surfaces on monolayer formation and barrier integrity of human umbilical vein endothelial cells (HUVEC) was then assessed by confocal imaging of the adherens junctional molecule VE cadherin, perijunctional actin and permeability to 42kDa dextrans. Dichlorofluoroscein assays were used to measure oxidative stress in the cells. XPS analysis of NiTi revealed its surface to be dominated by TiO(2). However, where oxidation had occurred after mechanical polishing or post polishing heat treatments at 300 and 400 degrees C in air, a significant amount of metallic nickel or nickel oxide species (10.5 and 18.5 at%) remained on the surface. Exposure of HUVECs to these surfaces resulted in increased oxidative stress within the cells, loss of VE-cadherin and F-actin and significantly increased paracellular permeability. These pathological phenomena were not found in cells grown on NiTi which had undergone heat treatment at 600 degrees C. At this temperature thickening of the TiO(2) layer had occurred due to diffusion of titanium ions from the bulk of the alloy, displacing nickel ions to sub-surface areas. This resulted in a significant reduction in nickel ions detectable on the sample surface (4.8 at%). This study proposes that the integrity of human endothelial monolayers on NiTi is dependent upon the surface chemistry of the alloy and that this can be manipulated, using simple oxidising heat treatments. PMID- 15814135 TI - Pharmacokinetic behaviour of ACP gel, an autocrosslinked hyaluronan derivative, after intraperitoneal administration. AB - Autocrosslinked polysaccharide (ACP) gel is a fully biocompatible cross-linked derivative of hyaluronic acid, which has prolonged in vivo residence time and improved mechanical properties with respect to native hyaluronan for use in various surgical applications. The objective of this study was to assess the pharmacokinetic behaviour of ACP gel in dogs after intraperitoneal administration. Seven beagle dogs received intraperitoneal injections of tritium labelled ACP gel. Blood samples were taken, and urine and faeces were collected until sacrifice, scheduled at various time points from 3 to 192 h after administration. Organs were removed from the animals at autopsy. Bodily fluid and organ samples were analysed for total and non-volatile radioactivity. Non volatile radioactivity slowly appeared in plasma, with a median T(max) of 12 h, and then declined with a mean half-life of 69 h. Total radioactivity in plasma peaked later and declined more slowly, consistent with the formation of tritiated water. Little non-volatile radioactivity was found in any organs except the liver, where about 16% of the dose was present 72 h after administration, and the intestines, where the presence of radioactivity was probably due to a retention effect. A minor amount of non-volatile radioactivity was also found in the bone marrow. In summary, ACP gel administered into the peritoneal cavity is removed slowly by active initial catabolism at the injection site, and is then catabolised by well described physiological pathway of hyaluronan degradation with final release of simple molecules such as CO(2) and H(2)O. Given its in vivo residence time, ACP gel may be considered an ideal implantable surgical device. PMID- 15814136 TI - Biodegradable dextran-based microspheres for delivery of anticancer drug mitomycin C. AB - The purpose of this work was to develop a biodegradable microsphere (MS) system for delivering mitomycin C (MMC). Various dextran-based MS systems were investigated for their loading and release characteristics, including nonionic MS, sulfopropyl dextran microspheres (SP-MS) with low or high cross-linking density, oxidized SP-MS (Ox-MS), and hydrophobically modified SP-MS. SP-MS were chemically modified by oxidation with sodium periodate or by reaction with anhydride. The chemical structure of modified SP-MS and MMC-loaded MS (MMC-MS) were examined using Fourier transform infrared (FTIR) and solid-state nuclear magnetic resonance (NMR) spectrophotometry. Drug release was conducted at 37 degrees C in aqueous solutions of 0.15 m phosphate buffer solution. The kinetics of drug absorption and release and the stability of MMC after loading and release were determined by spectrophotometry and high-performance liquid chromatography. Ionic SP-MS exhibited a higher drug-loading rate and capacity when compared to nonionic MS, while hydrophobically modified SP-MS showed an even greater loading capacity than SP-MS. These results suggest that both ionic complexation and hydrophobic interaction were important factors in MMC loading. The Ox-MS system demonstrated higher drug-loading capacity, more fractional drug release and a longer time to reach release equilibrium as compared to other investigated MS systems. Under optimized reaction and loading conditions, MMC released from Ox-MS was found to be unaltered. This work demonstrates that the Ox-MS system is a potentially useful system for the delivery of MMC. PMID- 15814137 TI - Local perivascular delivery of anti-restenotic agents from a drug-eluting poly(epsilon-caprolactone) stent cuff. AB - The introduction of drug-eluting stents (DES) to prevent in-stent restenosis is one of the major advances in interventional cardiology. Currently many types of DES are under evaluation for effectiveness and safety, a time-consuming and difficult procedure in humans. An animal model that allows rapid evaluation of the present and upcoming therapeutic approaches to prevent in-stent restenosis is most valuable and still lacking. Here, a perivascular cuff to induce restenosis was constructed of a poly(epsilon-caprolactone) (PCL) formulation suitable for the controlled delivery of drugs. Placing the PCL cuff around the femoral artery, in vivo, resulted in reproducible restenosis-like lesions containing predominantly smooth muscle-actin positive cells. Loading the cuff with the anti restenotic compounds paclitaxel and rapamycin resulted, in vitro, in a sustained and dose-dependent release for at least 3 weeks. Paclitaxel- and rapamycin eluting PCL cuffs placed around the femoral artery of mice in vivo significantly reduced intimal thickening by 76 +/- 2% and 75 +/- 6%, respectively, at 21 days. Perivascular sustained release of both anti-restenotic agents is restricted to the cuffed vessel segment with no systemic adverse effects or effect on cuffed contralateral femoral arteries. Drug-eluting PCL cuffs provide an easy and rapid tool to evaluate anti-restenotic agents to be used in combination with the DES strategies. PMID- 15814138 TI - Fabrication of a cell array on ultrathin hydrophilic polymer gels utilising electron beam irradiation and UV excimer laser ablation. AB - Most of the surface patterning methods currently applied are based on lithography techniques and microfabrication onto silicon or glass substrates. Here we report a novel method to prepare patterned surfaces on polystyrene substrates by grafting ultrathin cell-repellent polymer layers utilising both electron beam (EB) polymerisation and local laser ablation techniques for microfabrication. Polyacrylamide was grafted onto tissue culture polystyrene (TCPS) dishes using EB irradiation. Water contact angles for these PAAm-grafted TCPS surfaces were less than 10 degrees (costheta = 0.99) with PAAm grafted amounts of 1.6 microg/cm(2) as determined by ATR/FT-IR. UV excimer laser (ArF: 193 nm) ablation resulted in the successful fabrication of micropatterned surfaces composed of hydrophilic PAAm and hydrophobic basal polystyrene layers. Bovine carotid artery endothelial cells adhered only to the ablated domains after pretreatment of the patterned surfaces with 15 microg/mL fibronectin at 37 degrees C. The ablated domain sizes significantly influenced the number of cells occupying each domain. Cell patterning functionality of the patterned surfaces was maintained for more than 2 months without loss of pattern fidelity, indicating that more durable cell arrays can be obtained compared to those prepared by self-assembled monolayers of alkanethiols, as described in previous reports. The surface fabrication techniques presented here can be utilised for the preparation of cell-based biosensors as well as tissue engineering constructs. PMID- 15814139 TI - Nanopattern-induced changes in morphology and motility of smooth muscle cells. AB - Cells are known to be surrounded by nanoscale topography in their natural extracellular environment. The cell behavior, including morphology, proliferation, and motility of bovine pulmonary artery smooth muscle cells (SMC) were studied on poly(methyl methacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS) surfaces comprising nanopatterned gratings with 350 nm linewidth, 700 nm pitch, and 350 nm depth. More than 90% of the cells aligned to the gratings, and were significantly elongated compared to the SMC cultured on non-patterned surfaces. The nuclei were also elongated and aligned. Proliferation of the cells was significantly reduced on the nanopatterned surfaces. The polarization of microtubule organizing centers (MTOC), which are associated with cell migration, of SMC cultured on nanopatterned surfaces showed a preference towards the axis of cell alignment in an in vitro wound healing assay. In contrast, the MTOC of SMC on non-patterned surfaces preferentially polarized towards the wound edge. It is proposed that this nanoimprinting technology will provide a valuable platform for studies in cell-substrate interactions and for development of medical devices with nanoscale features. PMID- 15814140 TI - Size-controlled hydroxyapatite nanoparticles as self-organized organic-inorganic composite materials. AB - This paper presents some results concerning the size-controlled hydroxyapatite nanoparticles obtained in aqueous media in a biopolymer matrix from soluble precursors salts. Taking the inspiration from nature, where composite materials made of a polymer matrix and inorganic fillers are often found, e.g. bone, shell of crustaceans, shell of eggs, etc., the feasibility on making composite materials containing chitosan and nanosized hydroxyapatite was investigated. A stepwise co-precipitation approach was used to obtain different types of composites by means of different ratio between components. The synthesis of hydroxyapatite was carried out in the chitosan matrix from calcium chloride and sodium dihydrogenphosphate in alkaline solutions at moderate pH of 10-11 for 24 h. Our research is focused on studying and understanding the structure of this class of composites, aiming at the development of novel materials, controlled at the nanolevel scale. The X-ray diffraction technique was employed in order to study the kinetic of hydroxyapatite formation in the chitosan matrix as well as to determine the HAp crystallite sizes in the composite samples. The hydroxyapatite synthesized using this route was found to be nano-sized (15-50 nm). Moreover, applying an original approach to analyze the (002) XRD diffraction peak profile of hydroxyapatite by using a sum of two Gauss functions, the bimodal distribution of nanosized hydroxyapatite within the chitosan matrix was revealed. Two types of size distribution domains such as cluster-like (between 200 and 400 nm), which are the habitat of ''small'' hydroxyapatite nanocrystallites and scattered-like, which are the habitat of ''large'' hydroxyapatite nanocrystallites was probed by TEM and CSLM. The structural features of composites suggest that self-assembly processes might be involved. The composites contain nanosized hydroxyapatite with structural features close to those of biological apatites that make them attractive for bone tissue engineering applications. PMID- 15814141 TI - Microarray analysis of fluoro-gold labeled rat dopamine neurons harvested by laser capture microdissection. AB - The cellular heterogeneity of brain tissue presents a challenge to gene expression profiling of specific neuronal cell types. The present study employed a fluorescent neural tracer to specifically label midbrain dopamine neurons and non-dopamine cortical neurons. The labeled cells were then used to visually guide harvesting of the cells by laser capture microdissection (LCM). RNA extracted from the two populations of harvested cells was then amplified, labeled and co hybridized to high density cDNA microarrays for two-color differential expression profiling. Many of the genes most highly enriched in the dopamine neurons were found to be genes previously known to define the dopamine neuronal phenotype. However, results from the microarray were only partially validated by quantitative RT-PCR analysis. The results indicate that LCM harvesting of specific neuronal phenotypes can be effectively guided in a complex cellular environment by specific pre-labeling of the target cell populations and underlie the importance of independent validation of microarray results. PMID- 15814142 TI - Caudal thoracic air sac cannulation in zebra finches for isoflurane anesthesia. AB - Small songbirds such as the zebra finch are commonly used for studies on the neural mechanisms that underlie vocal learning. For these studies, survival surgeries are often performed that involve animal anesthesia and stereotaxic stabilization for localization of specific brain regions. Here we describe air sac cannulation as a novel method for delivering isoflurane gas to zebra finches for anesthesia during neurosurgery. Advantages of this method include that it leaves the bird's head free for stereotaxic targeting and does not interfere with the beak clamps that are often used to position and stabilize the head. It additionally allows for the use of the inhalant anesthetic, isoflurane, which is an appealing alternative to injectable anesthetics because it provides fast, minimally stressful induction, and low subject and personnel toxicity. The use of isoflurane also prevents overdosing and lengthy postoperative recovery times. PMID- 15814143 TI - Behavioral analysis during the forced swimming test using a joystick device. AB - The behavioral test described by Porsolt in 1977 for screening potential antidepressant drugs is extensively used both in basic research and in the pharmaceutical industry. The measured behavior is the immobility time during the swimming test (preformed in rodents), which decreases upon acute antidepressant treatment. Several research groups have suggested some modifications on the original Porsolt paradigm and its analysis. Nevertheless, there are still inaccuracies resulting from either undefined intermediate behaviors or from considering the movement of the whole body as one unit without analyzing the motion of the limbs. Herein, we propose a novel and simple scoring method, based on continuous measurement of the limbs motion, using a joystick, a computer screen and simple software. We validated the method, using antidepressant drugs and studied examples of false positives and false negatives of the traditional Porsolt paradigm. The proposed method is easy to use, it accounts for all range of movements and the analysis is relatively fast. Moreover, the results obtained using this analysis method show a normal Gaussian distribution in a population of rats (while the traditional Porsolt analysis does not) which allows selective breeding of 'motivated' and 'depressed' lines of animals. PMID- 15814144 TI - Validation of a digital video tracking system for recording pig locomotor behaviour. AB - We are introducing a system for automatically tracking pig locomotor behaviour. Transposing methods for the video-based tracking of rodent behaviour engenders several problems. We have therefore improved existing methods, based on image subtraction, to offer increased flexibility and accuracy in tracking large-sized animals in situations with a constantly changing background. The improved tracking algorithms introduce a reference frame, which does not include the animal and is automatically updated, and implementation of an automatic threshold detection algorithm. This makes the system more robust to the tracking environment, which could even be of the same colour as the animal, and allows the tracking environment to change during recording. We validated the system by estimating the repeatability, accuracy, and basic noise level, and tested the system in different levels of animal activity evoked by administration of apomorphine (APO) to minipigs in an open field test. Seven pigs each received the vehicle and three doses of APO (0.05, 0.1, and 0.3 mg/kg i.m.), and the locomotor behaviour of each session was recorded for 60-min. The calculated coefficient of repeatability was 0.6%, indicating high repeatability and the basic noise level of the tracking system was estimated to be 2%. Administration of the two lowest doses of APO was accompanied by increased locomotor activity of the pigs. Thus, this digital video-based tracking system for automatically tracking the spontaneous locomotor behaviour of pigs is highly reliable and accurate, and was able to detect well-known effects of APO in pig locomotor activity. PMID- 15814145 TI - Influence of high motor unit synchronization levels on non-linear and spectral variables of the surface EMG. AB - The aim of this study was to investigate the influence of high degrees of motor unit synchronization on surface EMG variables extracted by linear and non-linear analysis techniques. For this purpose, spectral and recurrent quantification analysis (RQA) were applied to both simulated and experimental EMG signals. Synthetic surface EMG signals were generated with a model of volume conductor comprising muscle, fat, and skin tissues. The synchronization was quantified by the percent of discharges of each motor unit synchronized with discharges of other motor units. The simulated signals presented degrees of synchronization in the range 0-80% (10% increments) and three mean values of motor unit conduction velocity distribution (3, 4 and 5 m/s). Experimental signals were collected from the first dorsal interosseous muscle of five patients with Parkinson disease during 10s of rest and 10s of isometric voluntary contraction at 50% of the maximal force. Mean power spectral frequency (MNF) and percent of determinism (%DET) of the surface EMG were computed from the simulated and experimental signals. In the simulated signals, %DET was linearly related to the level of synchronization in the entire range considered while MNF was sensitive to changes in synchronization in a smaller range (0-20%), outside which it levelled off. The experimental results indicated that %DET was significantly higher in the resting condition (with presence of tremor; mean +/- S.E., 85.4 +/- 0.8%) than during the voluntary contraction (which partly suppressed tremor; 60.0 +/- 2.3%; P < 0.01). On the contrary, MNF did not depend on the condition (114.3 +/- 1.5 Hz and 118.0 +/- 0.8 Hz for the resting and voluntary contraction, respectively), confirming the simulation results. Overall, these results indicated that linear and non linear analyses of the surface EMG may have different sensitivities to the underlying physiological mechanisms in specific conditions, thus their joint use provides a more complete view of the muscle status than spectral analysis only. PMID- 15814146 TI - Inexpensive system for real-time 3-dimensional video-oculography using a fluorescent marker array. AB - We describe a novel, inexpensive method for real-time measurement of binocular three-dimensional eye position. The method employs consumer-grade digital video cameras ("webcams") to track an array of three fluorescent non-collinear markers affixed to each eye. The instantaneous position of the marker array relative to a reference position is used to construct a rotation matrix describing the eye rotation. The mathematical computation used to determine the rotation matrix is conceptually simpler and computationally more efficient than methods previously described, allowing generation of binocular three-dimensional eye position in real-time during image acquisition. The fluorescent marker is illuminated using a UV-A light source. The light source and reflective artifacts are filtered out to improve the signal to noise ratio. In addition, we present a method to align the camera with the center of eye rotation. When tested in vitro, the video oculography (VOG) method had a <2.9% positional error (in each component of 3-D eye position) for eye positions within 20 degrees of center. We directly compared this method of VOG to the search coil technique by measuring three-dimensional eye position simultaneously using search coils and VOG in a chinchilla (C. laniger). The in vivo positional difference between the two methods was <3.1% for each component of 3-D eye position. PMID- 15814147 TI - Targeted gene delivery to telencephalic inhibitory neurons by directional in utero electroporation. AB - Telencephalic inhibitory neurons originate in the ganglionic eminences and migrate to the cerebral cortex following a tangential trajectory, before they differentiate and integrate within the local circuitry. Current studies of interneuron development and function benefit from the use of knock-out and transgenic mice, whereas none take advantage of the versatility of in utero electroporation. Here, we show how in utero electroporation can be directed to the ganglionic eminences to specifically target gene expression to interneurons. Electroporation of GFP-encoding plasmids into the ganglionic eminences results in selective labeling of migrating interneurons during development. In the adult brain of electroporated animals, a wide variety of cortical, hippocampal and olfactory bulb interneurons are labeled. We also show that GFP-expressing interneurons can be visualized in living slices of adult cerebral cortex, where they display normal electrophysiological properties. Photostimulation studies using acute slices show that cortical GFP+ interneurons receive normal, layer specific synaptic input, indicating that these neurons integrate within the local cortical circuitry. Ganglionic eminence-directed in utero electroporation is therefore an effective, rapid, and versatile method of selectively transfecting telencephalic interneurons, optimal for both developmental studies and adult functional studies. PMID- 15814148 TI - An evaluation of the use of seizure prone rats when investigating intermediate stage sleep. AB - A body of literature is developing which identifies an additional stage of sleep in rats, cats and mice. Intermediate stage (IS) sleep is a measurable sleep stage that is maintained by the hyperpolarization of GABA(A) containing thalamocortical neurons. The present study attempts to clarify inconsistencies within the sleep spindle literature. Most notably, inconsistencies between those that study sleep spindles in the rat outside and within the context of IS sleep. Ten male taiep rats weighing from 400 g to 600 g, and 9-12 months of age, were used in this study. The animals were given a one-time, .9 mg/kg dose of the benzodiazepine clonazepam. The control group had more seizure activity (mean = 13.4) than the treatment group (mean = 5.2, t(1-18) = 8.859, p < .001), and had a lower number of sleep spindles (mean=10.3) than the treatment group (mean = 13.3, t(1-18) = 3.4, p < .001). In addition, spectral analysis of sleep spindles during IS and seizure activity revealed that sleep spindles are within the frequency band of 8 11Hz, while seizure activity is within the 4-7 Hz range. This data supports the hypotheses that sleep spindles are distinguishable from seizure activity. PMID- 15814149 TI - A novel ELISA using PVDF microplates. AB - Here we describe the development of a novel specific, rapid ELISA system, which is performed on modified microplates where polyvinylidine fluoride (PVDF) forms the base of each well. The use of microplates with PVDF membranes as the solid phase allows for a greater binding capacity of protein in comparison to the solid phases of traditional ELISAs. The increased binding capacity of the solid phase provides for the direct binding of antigens, which can subsequently be assayed using a single, specific and well-characterized antibody. This direct assay system eliminates the need for two distinct antibodies that are often necessary in conventional two site ELISA systems. The system is able to specifically detect purified proteins as well as antigens in crude preparations of tissue homogenates. The PVDF-based ELISA performs with similar sensitivity and reproducibility as conventional two site ELISAs in tissue homogenates. The intra- and inter-assay coefficients of variation for the measurement of actin in crude rat brain homogenate were 2.36 and 5.15%, respectively. PMID- 15814150 TI - The grooming analysis algorithm discriminates between different levels of anxiety in rats: potential utility for neurobehavioural stress research. AB - Stress has long been known to affect grooming in rodent species, altering both its activity measures and behavioural microstructure. Since stress disturbs a general pattern of self-grooming uninterrupted cephalocaudal progression, the grooming analysis algorithm (Kalueff and Tuohimaa, Brain Res. Protocols, 2004; 15: 151-8) was previously designed for mice to enable the detection of stress by measuring alterations in grooming microstructure in different test situations. Since mice and rats are known to differ in their behaviours, including grooming, the aim of the current study was to test our approach in rats and evaluate the utility of this method for differentiation between high- and low-stress situations. For this, we have developed the rat grooming analysis algorithm (based on ethological analysis of incorrect transitions contrary to the cephalocaudal rule, interrupted grooming activity and the assessment of the regional distribution of grooming) and applied this algorithm to the light exposed (high stress) and dark-exposed (low stress) groups of rats. Here, we show that the percentage of 'incorrect' transitions between different grooming patterns, the percentage of interrupted grooming bouts and altered regional distribution of grooming (less caudal grooming, more rostral grooming) may be used as behavioural markers of stress in rats. Our results suggest that this method can be a useful tool in neurobehavioural stress research including modelling stress-evoked states, psychopharmacological or behavioural neurogenetics research in rats. PMID- 15814151 TI - Methods for functional magnetic resonance imaging in normal and lesioned behaving monkeys. AB - Methods for performing functional magnetic resonance imaging (fMRI) studies in behaving and lesioned monkeys using a human MR scanner are reported. Materials for head implant surgery were selected based on tests for magnetic susceptibility. A primate chair with a rigid head fixation system and a mock scanner environment for training were developed. To perform controlled visual studies, monkeys were trained to maintain fixation for several minutes using a novel training technique that utilized continuous juice rewards. A surface coil was used to acquire anatomical and functional images in four monkeys, one with a partial lesion of striate cortex. High-resolution anatomical images were used after non-uniform intensity correction to create cortical surface reconstructions of both lesioned and normal hemispheres. Our methods were confirmed in two visual experiments, in which functional activations were obtained during both free viewing and fixation conditions. In one experiment, face-selective activity was found in the fundus and banks of the superior temporal sulcus and the middle temporal gyrus in monkeys viewing pictures of faces and objects while maintaining fixation. In a second experiment, regions in occipital, parietal, and frontal cortex were activated in lesioned and normal animals viewing a cartoon movie. Importantly, in the animal with the striate lesion, fMRI signals were obtained in the immediate vicinity of the lesion. Our results extend those previously reported by providing a detailed account of the technique and by demonstrating the feasibility of fMRI in monkeys with lesions. PMID- 15814152 TI - Analysis of the time-varying energy of brain responses to an oddball paradigm using short-term smoothed Wigner-Ville distribution. AB - Cognitive brain responses to external stimuli, as measured by event related potentials (ERPs), have been analyzed from a variety of perspectives to investigate brain dynamics. Here, the brain responses of healthy subjects to auditory oddball paradigms, standard and deviant stimuli, recorded on an Fz electrode site were studied using a short-term version of the smoothed Wigner Ville distribution (STSW) method. A smoothing kernel was designed to preserve the auto energy of the signal with maximum time and frequency resolutions. Analysis was conducted mainly on the time-frequency distributions (TFDs) of sweeps recorded during successive trials including the TFD of averaged single sweeps as the evoked time-frequency (ETF) brain response and the average of TFDs of single sweeps as the time-frequency (TF) brain response. Also the power entropy and the phase angles of the signal at frequency f and time t locked to the stimulus onset were studied across single trials as the TF power-locked and the TF phase-locked brain responses, respectively. TFDs represented in this way demonstrated the ERP spectro-temporal characteristics from multiple perspectives. The time-varying energy of the individual components manifested interesting TF structures in the form of amplitude modulated (AM) and frequency modulated (FM) energy bursts. The TF power-locked and phase-locked brain responses provoked ERP energies in a manner modulated by cognitive functions, an observation requiring further investigation. These results may lead to a better understanding of integrative brain dynamics. PMID- 15814153 TI - Beyond subgroup analysis: improving the clinical interpretation of treatment effects in stroke research. AB - In large clinical trials designed to determine efficacy of an experimental treatment, patients are enrolled with presence or absence of various risk factors, such as diabetes or history of atrial fibrillation. A treatment-by-risk factor interaction indicates that the treatment effect may depend on the risk factor presence or absence. It is important to identify such interaction, since a treatment may fail or cause adverse events in the presence of the risk. Although statistical methods exist to identify such interaction, they are underutilized in clinical stroke research. This paper reviews the notion of treatment-by-risk factor interaction and identifies two types of interaction, quantitative and qualitative, using a graphical technique and statistical testing. We illustrate how to avoid drawing the erroneous conclusions regarding the treatment effect on subgroups when failing to detect an interaction, and provide rigorous tools to estimate the treatment effect on subgroups when an interaction is observed. Applications are presented using the data collected from the NINDS t-PA stroke studies. In stroke clinical trials, a treatment-by-risk factor interaction must be considered if the data permit. The graphical approach provides a heuristic illustration of interactions. Qualitative interactions are more important than quantitative interactions on therapeutic conclusion. Results of NINDS t-PA stroke studies confirmed our previous conclusions on the treatment t-PA benefit within 3 h therapeutic window. No subgroup of patients would lead a physician to withhold the t-PA treatment. PMID- 15814154 TI - Norepinephrine transporter function and desipramine: residual drug effects versus short-term regulation. AB - Previous research has shown that exposure of norepinephrine transporter (NET) expressing cells to desipramine (DMI) downregulates the norepinephrine transporter, although changes in the several transporter parameters do not demonstrate the same time course. Exposures to desipramine for <1 day reduces only radioligand binding and uptake capacity while transporter-immunoreactivity is unaffected. Recent demonstration of persistent drug retention in cells following desipramine exposures raises the possibility that previous reported changes in the norepinephrine transporter may be partly accountable by residual drug. In this study, potential effects of residual desipramine on norepinephrine transporter binding and uptake were re-evaluated following exposures of PC12 cells to desipramine using different methods to remove residual drug. Using a method that minimizes residual drug, exposure of intact PC12 cells to desipramine for 4h had no effect on uptake capacity or [(3)H]nisoxetine binding to the norepinephrine transporter, while exposures for > or =16 h reduced uptake capacity. Desipramine-induced reductions in binding to the transporter required >24 h or greater periods of desipramine exposure. This study confirms that uptake capacity of the norepinephrine transporter is reduced earlier than changes in radioligand binding, but with a different time course than originally shown. Special pre-incubation procedures are required to abolish effects of residual transporter inhibitor when studying inhibitor-induced transporter regulation. PMID- 15814155 TI - In situ immunoradiographic method for quantification of specific proteins in normal and ischemic brain regions. AB - This study tested the application of an immunoisotopic assay for immunohistochemical localization and quantification of proteins in brain sections from rats without or with transient focal ischemia. We assessed the hypothesis that measurements of protein levels in injured brain determined by an isotopic assay using [(125)I]-protein A have greater reliability than those made by conventional immunoperoxidase labeling using diaminobenzidine. Quantification of immunoreactivities for glial fibrillary acidic protein (GFAP), glutamate transporter-1 (GLT-1) and heat shock protein-70 (HSP-70) was determined by optical density signal in the immunoisotopic and immunoperoxidase assays. In ischemic brain, the immunoisotopic assay detected protein increases (cortical penumbra HSP-70, 151+/-6%), protein decreases (cortical ischemic core GLT-1, 61+/ 6%) and no changes in GFAP levels compared to controls animals. These results differed from the protein levels found by the immunoperoxidase assay, which showed elevated HSP-70, GLT-1 and GFAP in all ischemic regions. We conclude that nonspecific immunosignal confounds assessments of protein expression in injured brain and that the immunoisotopic method is a valid approach to regionally localize and quantify proteins after brain injury. The disadvantage of the falsely positive overestimation of protein immunoreactivity after stroke with the immunoperoxidase method has to be weighted with the advantage of the cellular resolution. PMID- 15814156 TI - Aging changes in the periodontal bone of F344/N rat. AB - The aim of this research was to determine whether a rat was an adequate laboratory animal model for periodontal research on elderly humans. Thirty-two F344/NSlc female rats ranged between 30 and 1000 days of age were used. The alveolar bone loss around the molars was assessed by a morphometric method. A significant correlation was found between age and the amount of alveolar bone loss. For further analysis, the rats were grouped into four by age; 30-60 days, 220-430 days, 640-850 days, and more than 850 days. The means of alveolar bone loss were compared between age groups. It was found that the resorption of the alveolar bone around the molars of the rats continued until they were 1000-days old, and this trend was stronger in the mandible than the maxilla. It was suggested that rats could be used as adequate laboratory animals for periodontal research. PMID- 15814158 TI - Successful aging in centenarians: myths and reality. AB - The term "successful aging" appeared in the first issue of "The Gerontologist" in 1961. During the successive years, this expression has changed its meaning. Nowadays, successful aging means "absence of diseases and disabilities, maintenance of high levels of physical and cognitive abilities, preservation of the social and productive activities". It has become a common opinion that the centenarians may represent the prototypes of the successful aging. This motivated our work to study the clinical, psychical, and functional aspects in a centenarian group, verifying the real autonomy, instrumental capacities, and working abilities. Our study pool consisted of 602 centenarians, who were also subjects of then epidemiological studies of the Italian Multicenter Studies on Centenarians (IMUSCE). All subjects underwent a clinical-anamnestic evaluation, cognitive-functional tests by means of the mini mental state examination (MMSE), the independence index in activities of daily living (ADL), the instrumental activities of daily living (IADL) scale. The centenarians were classified in three groups, according to the criteria elaborated previously by us, based on their psychophysical status and autonomy, as follows. Group A: centenarians in good health status; Group B: centenarians in an intermediate health status. Group C: centenarians in bad health status. Group A represented 20.0% of the total pool, Group B amounted to 33.4%, and the Group C was 46.6%. The centenarians of Group A presented normal ADL values, and 47.9% of them were autosufficient in all functions; 5.7% of them were independent in all IADL items. These data confirm that the centenarians of Group A are free of invalidating chronic diseases, are autonomous, maintain good physical and cognitive capacities, however, have not maintained any social or productive activities. Therefore, they cannot be considered as prototypes of successful aging. PMID- 15814157 TI - Prooxidant activities of alpha-lipoic acid on oxidative protein damage in the aging rat heart muscle. AB - In the present study, we investigated whether alpha-lipoic acid (ALA) supplementation could have prooxidant or antioxidant effects on protein oxidation parameters such as protein carbonyl (PCO), nitrotyrosine (NT), advanced oxidation protein products (AOPP), and protein thiol (P-SH), as well as oxidative stress parameters such as total thiol (T-SH), non-protein thiol (Np-SH), and lipid hydroperoxide (LHP) in the heart muscle tissue of aged rats. ALA (100 mg/kg body wt/day) was administered intraperitoneally to the experimental animals for 14 days. PCO, NT, AOPP, and P-SH levels were increased, T-SH and Np-SH levels were not changed, and only LHP levels were decreased in the heart muscle tissue of aged rats with ALA supplementation. When compared with non-supplemented aged rats, increasing levels of protein oxidation markers such as PCO, NT, and AOPP in ALA-supplemented aged rats may suggest that oxidative protein damage is increased in ALA-supplemented aged rats. We assume that an explanation for our findings regarding ALA supplementation on protein oxidation markers in the heart muscle tissue of aged rats may be due to the prooxidant effects of ALA. The prooxidant effects of ALA supplementation should be considered in future studies. PMID- 15814159 TI - The vestibular dysfunction and anxiety disorder interface: a descriptive study with special reference to the elderly. AB - Vestibular abnormalities co-existing with anxiety disorders are not uncommon and there has been a renewal of interest in recent times. Although well known over centuries, there is often a delay in the recognition of this relationship by the primary care physician and the specialist alike. Dizziness embracing vertigo, unsteadiness and imbalance are common in the elderly, so is generalized anxiety disorder, which is a common psychiatric problem in later life. This is a retrospective study of eight patients with vestibular symptoms and an anxiety disorder present over several years with lack of awareness of their relationship. The diagnoses of the anxiety disorders were based on the Diagnostic and Statistical Manual (DSM-IV) criteria and the effect of treatment measured on a clinician-based impression interview. There was one male and seven females and the mean age was 72 years. Apart from the vestibular symptoms present in all the patients, the anxiety disorders comprised, generalized anxiety disorder in three, panic attacks in five and with agoraphobia in three. Four patients had hyperventilation, one sleep apnea, and two somatization disorders. They had all presented to clinicians in different disciplines and had had several investigations. Five had been treated in this study with alprazolam and three with citalopram, with modest to good results. Two had rehabilitation therapy as well. The cases described mirror the well-documented co-existence of vestibular and anxiety disorders together with hyperventilation and sleep apnea. The positive findings associated with vestibular dysfunction need recognition in addition to the non-specific psychiatric and behavioral symptoms. We emphasize this relationship and review the literature to alert the clinician. PMID- 15814160 TI - Cerebral white matter lesions and hypertension status in the elderly Korean: the Ansan Study. AB - There is evidence of an association of hypertension with white matter lesions (WMLs), leukoaraiosis, in the human brain in the United States, Europe and Japan. However, no study on this association has been reported in Korea. Community dwelling elderly subjects aged over 61 years were randomly selected from samples of the Ansan Health Study, while their blood pressures (BPs) were measured using a highly standardized protocol. Hypertension was defined as a systolic BP> or =140 mmHg or a diastolic BP> or =90 mmHg or as reported treatment with hypertensive medication, and further subclassified with respect to its subtypes and control status. WMLs were determined by a 1.5-T MRI scan and further categorized into five subgroups with respect to its severity. Hypertension status, such as uncontrolled hypertension even after antihypertensive medication (adjusted odd ratio [OR]=6.07; 95% CI, 2.08-17.78), isolated systolic hypertension before medication (OR=2.66; 95% CI, 1.08-6.57), and hypertension before medication (OR=2.55; 95% CI, 1.19-5.46) were significantly and independently correlated with the presence and severity of WMLs. Advancing age (OR=1.76; 95% CI, 1.38-2.25), lacunar infarction (OR=3.19; 95% CI, 1.83-5.58), and cigarette smoking (OR=1.10; 95% CI, 1.03-1.18) were significantly related to the presence and severity of WMLs. Regular and tight control for hypertension should be exercised to prevent WMLs among the elderly with hypertension. PMID- 15814161 TI - An evaluation of efficacy and safety of reboxetine in elderly patients affected by "retarded" post-stroke depression. A random, placebo-controlled study. AB - Depression occurs frequently in post-stroke patients and appears to be associated with an impairment in their rehabilitation and functional recovery. Although selective serotonin reuptake inhibitors (SSRI) are often used in post-stroke depression (PSD), it has been observed that only a subset of patients is responsive to this treatment. Other patients respond to tricyclic antidepressants or MAO inhibitors, which, however, may not have a favorable profile of safety and tolerability in post-stroke patients. In this double-blinded, placebo-controlled study, we evaluated the efficacy and tolerability of the noradrenaline reuptake inhibitor, reboxetine, in a subset of PSD patients classified as affected by "retarded" depression. Reboxetine (4 mg, twice daily, for 16 weeks) was administered to patients that developed depression after a single ischaemic or hemorrhagic stroke. We assessed the severity of depressive symptoms by the Beck Depression Inventory (BDI) and Hamilton Depression Rating Scale (HDRS). HDRS and BDI scores (mean+/-S.D.) at baseline were, respectively, 24+/-1.31 and 19.87+/ 1.46 in the placebo group, 24.06+/-1.52 and 20.56+/-2.16 in the reboxetine group. After 16 weeks, HDRS and BDI mean scores were respectively 22.73+/-2.4 and 18.4+/ 3.33 in the placebo group, 9.26+/-2.15 and 8.06+/-3.43 in the reboxetine group [p<0.01 versus the respective baseline (paired t-test); (#)p<0.01 versus retarded depressed patients treated with placebo (one-way analysis of variance (ANOVA) applied to the difference from baseline, associated with Dunnett's t-test to isolate the differences)]. Reboxetine showed a good efficacy, safety and tolerability in PSD patients affected by "retarded" depression. We conclude that reboxetine is well tolerated and may be a useful therapeutic option in PSD patients with "retarded" depression. PMID- 15814162 TI - A population-based study on well-being in the very old: the role of cardiovascular diseases and drugs. AB - Cardiovascular diseases constitute the most common health problems in very old people. Consequently, cardiovascular drugs are the medicines that are most frequently used by elderly subjects. Although many studies have examined the physiological effect and adverse reactions of these drugs, knowledge on their effect on emotional well-being is missing. The present study aims to examine the association between cardiovascular diseases and their medical treatment on the emotional well-being of very old people. We investigated a representative group of elderly subjects gathered from a population-based study (n=235). Participants were 84 years or older and cognitively intact (mini-mental state examination (MMSE) > or =24 points). Well-being was assessed with the positive and negative affect schedule (PANAS), measuring different mood categories. Cardiovascular diseases were diagnosed following the International Classification of Diseases. In this population the prevalence of cardiovascular diseases was high (62%). Multivariate regression analysis showed that while being affected by a cardiovascular disease did not affect the emotional well-being of the subjects (PANAS-PA, p=0.171; PANAS-NA, p=0.209), the use of some cardiovascular drugs showed an association. Cardiac glycosides (p=0.006) and nitrates (p=0.008) were associated with increased negative feelings. Due to high prevalence of cardiovascular diseases and use of cardiovascular medicines, this finding has relevance on the quality of life of elderly people. However, due to the nature of this study we cannot assess cause-effect relationship of this positive association. Therefore, the present findings suggest that there is a need for clinical studies in this increasing and limited studied age group. PMID- 15814163 TI - Interleukin-18 (IL-18) and matrix metalloproteinase-9 (MMP-9) in post-menopausal osteoporosis. AB - This survey covered 60 post-menopausal women with osteoporosis. The patients were divided into three equal groups, and each group was treated with one of the three so-called anti-resorptive drugs, namely alendronate (10 mg/day) risedronate (5 mg/day) and raloxifene (60 mg/day) for 12 months. The Elisa technique was used to measure circulating IL-18 and MMP-9. Lumbar bone mineral density (BMD) levels were determined by using dexa mineralometry (Lunar DPX) at baseline and after 12 months of treatment. The results showed comparable responses of the patients treated with alendronate or risedronate, being a significant increase in BMD, an increase in circulating IL-18, and only slight modifications in circulating MMP-9 levels. After 12 months of treatment with raloxifene, there were minimal, non significant increases in BMD, slight modifications in IL-18 levels, and a significant reduction in circulating MMP-9 levels. The conclusions can be drawn that all three drugs, albeit through different mechanisms, can be considered valid treatments for post-menopausal osteoporosis. Although measurements of circulating IL-8 and MMP-9 levels allowed us to differentiate the effects of the three drugs used, as of today, they have no real role in the diagnosis and/or follow-up of osteoporosis. PMID- 15814164 TI - Depression and concomitant diseases in a Turkish geriatric outpatient setting. AB - Depression is an important but inadequately diagnosed mood disorder in elderly. Depressed elderly patients often have chronic concomitant diseases. This paper intended to determine the prevalence of depression and its relation with concomitant disorders and social status among the patients admitted to our geriatric unit. Seven hundred and eighty-nine females and 466 males admitted to our unit were examined for the presence of depression by using the geriatric depression scale (GDS) test. The presence of concomitant diseases was assessed. Depression was diagnosed in 273 patients (21.8%), 193 (70.7%) females and 80 (29.3%) males. Depressed patients suffered from a wide range of other diseases the number and prevalence of which were as follows: Alzheimer's disease (AD) (34; 12.5%), vascular dementia (27; 9.9%), hypertension (HT) (211; 77.3%), diabetes mellitus (DM) (64; 23.4%), osteoporosis (182; 66.7%), atherosclerotic coronary artery disease (CAD) (89; 32.6%), cardiac failure (23; 8.5%), bronchial asthma (8; 2.9%), chronic obstructive pulmonary disease (COPD) (25; 9.2%) and osteoarthritis (133; 48.8%). The correlation between depression and concomitant diseases was statistically significant in hypertensive, demented and osteoporotic patients, as determined in a large elderly population. Previous studies examined the correlation of depression with only one concomitant disease, while we performed the analysis on multiple correlations. PMID- 15814165 TI - Water and sodium disturbances predict prognosis of acute disease in long term cared frail elderly. AB - Elderly patients are at increased risk for developing infections and fever. We aimed to investigate the incidence, causes and outcome of hypernatemia and hyponatremia among elderly long-term care patients with febrile illness. This is a prospective study conducted in a 110 beds, nursing department for dependent elderly in geriatric long-term care facility during 20 months period. Changes in plasma sodium concentrations were found in 48% (132/270) of the febrile illness: during the acute illness in 38% (102/270) of the events and during the follow-up period after acute febrile event, in 30% (59/198) of the events. Upper respiratory tract, bronchial, lung, and urinary tract infections were the most common causes for the fever. Hypernatremia was more prevalent on acute febrile illness (63%). Hyponatremia was more common before the febrile illness (10%) and during the follow-up period (69%). The most common causes of hypernatremia were inadequate hydration or correction (57%) and continuous diuretic treatment (38%). The most common causes of hyponatremia were: Syndrome of Inappropriate Antidiuretic Hormone Secretion (SIADH) (38%), incorrect hydration (19%), and continuous diuretic treatment (30%). During the study, 60% (28/47) of the patients died, and hypernatremia was detected in 82% (20/28). Disturbances in sodium concentrations were predictors of bad outcomes, related to the underlying diseases burden, and were not related to the magnitude of the changes in plasma sodium, time of its development, advanced age, gender and coexisting changes in plasma potassium level. PMID- 15814166 TI - Functional recovery after hip fracture in old-old elderly patients. AB - Hip fracture is epidemic and prevalence increased with advanced age. Impact of comorbid and cognitive status, gender, type of fracture, operative delay and pre fracture ambulatory levels on functional outcome was shown in previous studies. We studied functional outcome after rehabilitation for hip fracture in old-old elderly (85 years and older) and compared it to young elderly (65-74 years) community-dwelling patients. Before the fracture, old-old elderly patients were more functional dependent, has had more comorbid diseases, and more of them live alone than young elderly. The waiting time to surgery and mean length of stay in orthopedic ward were longer than in young elderly. On admission to rehabilitation treatment, old-old patients presented with more depressed mood, were more cognitive impaired, and more suffer from pain. Old-old patients presented with laboratory data of malnutrition (decreased serum levels of albumin, cholesterol, hemoglobin, hematocrit, lymphocyte count) and inflammation (increased of transferrin and C-reactive protein). Improvement in Functional Independence Measurement (FIM) scale was found in both groups but significantly better in young elderly than in old-old elderly. The change in FIM during the rehabilitation period (DeltaFIM) were in FIM total and in those parts of FIM concerning locomotion. The mean duration of rehabilitation stay was significantly longer in old-old elderly patients. On discharge old-old elderly patients more suffer from pain and difference between the groups according to the laboratory and to the cognitive data increased. Age per se is indicator of frailty and determinate functional recovery after hip fracture. PMID- 15814167 TI - Socio-economic position and its relationship to physical capacity among elderly people living in Jyvaskyla, Finland: five- and ten-year follow-up studies. AB - Socio-economic differences in self-reported disability are well described but much less is known about their associations with more objective measures of physical capacity. The aim was to study socio-economic differences in performance based physical capacity in 75-year-old persons, examining changes in performance at five- and ten-year follow-up intervals. At the baseline 350 residents of the city of Jyvaskyla, Finland, aged 75 were interviewed and 295 of them took part in clinical examinations. The corresponding figures at the five-year follow-up were 234 and 191 and at the ten-year follow-up 139 and 103. The statistical significance of differences in physical capacity between the socio-economic groups and genders were tested using ANOVA in univariate and repeated measures models and ANCOVA, with confounders added to the models. Generally, higher education and income were separately related to better maximal walking speed and vital capacity at every measurement point. In addition, higher income was related to better maximal isometric hand grip strength at both follow-ups. When education and income were in the same model, only income was related to physical capacity, almost without exception. Similarly, in the five- and ten-year follow-up periods, both education and income groups showed a parallel decline in physical capacity. The association between income and physical capacity remained even after adjusting for smoking, physical activity and number of chronic diseases. The results indicate that elderly people in disadvantaged socio-economic groups show lower levels of performance in almost all domains of physical capacity, but change in capacity over time does not differ significantly between either markers of socio-economic position. PMID- 15814168 TI - A model of underlying socioeconomic vulnerability in human populations: evidence from variability in population health and implications for public health. AB - Drawing from insights into the variability of complex biologic systems we propose that the health of human populations reflects the interrelationship between underlying vulnerabilities (determined by population-level social and economic factors; e.g., income distribution) and capacities (determined by population level salutary resources, e.g., social capital) and how populations, shaped by these vulnerabilities and capacities, respond to intermittent stressors (e.g., economic downturns) and protective events (e.g., introduction of a school). Monitoring this dynamic at the population-level can be accomplished by examining not only rates of illness and mortality, but variability in rates, either between populations or within populations over time. We used mortality data from New York City neighborhoods between 1990 and 2001 to test two related hypotheses consistent with this model of population health: (a) There is greater variability in mortality rates at a point in time between neighborhoods that are characterized by socioeconomic vulnerability; and (b) there is greater variability in mortality rates over time within neighborhoods that are characterized by socioeconomic vulnerability. We found that neighborhoods characterized by social and economic vulnerability displayed substantial variability in particular mortality rates. Mortality rates displaying the greatest variability were from causes that may be sensitive to social conditions (e.g., homicide or HIV/AIDS rates). Variability in population health existed both between neighborhoods with underlying vulnerability at one point in time and within vulnerable neighborhoods over time. The results of this analysis are consistent with a theory of underlying socioeconomic vulnerabilities of human populations and suggest that variability in population health may be an important consideration in population health assessment. PMID- 15814169 TI - A European comparative study of marital status and socio-economic inequalities in suicide. AB - We assess (i) whether being married is a protective factor against socio-economic inequalities in suicide, and (ii) whether any such buffering effect varies between countries. We used the data from a European cross-national comparison project, a prospective follow-up of several European population censuses matched with vital statistics. The data encompass 99.5 million person-years aged 30 and above and 25,476 suicides in Austria, Belgium, Denmark, Finland, Turin, Madrid, Norway and Switzerland. Standardised rates were computed and logistic regressions were used to assess educational inequalities. Among the non-married, the lower educational group had an increased risk of dying of suicide compared to the higher group (Odds Ratio (OR) = 1.45). Inequalities among the married were lower (OR = 1.29). In all countries or regions except Austria, the lower educational group had a higher risk of suicide mortality among the non-married than among the married. The buffering effect of being married was not observed for elderly individuals (65 and over). Among younger individuals, the buffering effect of being married on relative inequalities in suicide was stronger in Madrid, Denmark, Norway and Switzerland (but significant only for Denmark and Norway). There was no indication that countries with stronger welfare policies or lower divorce rate had a lower buffering effect. We conclude that being married protects against inequalities in suicide and that this protective effect is not affected by the level of social capital at the country level. PMID- 15814170 TI - A comparative study of the patterning of women's health by family status and employment status in Finland and Sweden. AB - The main aim of this study is to compare the patterning of health by family status and employment status among women in Finland and Sweden and to explore whether the patterning of health by family status is influenced by employment status and income. An additional aim was to identify which combinations of family status and employment status are especially disadvantageous for women's health . The data derived from comparable interview surveys carried out in 1994/1995. The analyses were restricted to ages 25-49; 2282 women in Finland and 2685 in Sweden. Firstly, age-adjusted prevalence percentages were presented by family status and employment status. Secondly, the patterning of health by family status and employment status, and the influence of adjusting for income, were studied by logistic regression analysis. The main results showed that women living in couples with children had the best health in both countries. Additional analyses showed that in Finland particularly poor health can be found among women who are simultaneously non-partnered and non-employed, whereas in Sweden poor health can be found among all non-employed groups of women. Income did not explain the poor health among the non-partnered and non-employed. This study confirmed that health is patterned by family status and employment status both among Finnish and Swedish women. It was found that non-employed women without a partner are likely to have poor health. In order to reduce inequalities in health among women, more efforts should be put on promoting health among these groups. PMID- 15814171 TI - The influence of neighborhood environment on the incidence of childhood asthma: a multilevel approach. AB - Some ecological analyses suggest an influence of neighborhood environment on asthma outcomes. However, no previous study has applied a multilevel approach to assess an ecological effect of neighborhood environment on the incidence of childhood asthma accounting for individual risk factors. This study assessed the influence of neighborhood and individual-level factors on the incidence of childhood asthma among all children born in Rochester, Minnesota, between 1976 and 1979. We identified asthmatics among all children born in Rochester, between 1976 and 1983. We applied a multilevel survival model with the frailty term to assess the effects of neighborhood characteristics, such as mean family income per census tract (n = 16) from the 1980 census report and the status of whether a census tract faces intersections with major highways or railroads, on asthma incidence. The relative risks (RR) of neighborhood socioeconomic status (SES), the status of whether census tracts face intersections with highways or railroads and the variance of random effect of census tracts were calculated adjusting individual-level covariates for asthma, including gender, birth weight, mother's age at birth and parental educational level at birth. We found that the RR of developing asthma among children living in census tracts facing intersections with highways or railroads was 1.6 (95% CI: 1.1-2.2) compared to those who lived in census tracts not facing intersections, adjusting individual- and neighborhood level covariates. The variance of the frailty term attributable to census tracts was small (0.0085) and was modified (from 0.004 to 0.0085, 112% change) by adding neighborhood covariates. The overall effects of individual-level factors on asthma incidence were independent of neighborhood environment. The influence of neighborhood environment on childhood asthma in a non-inner-city setting, like Rochester, Minnesota, was small to modest. Incorporating pertinent neighborhood level covariates into multilevel models needs to be considered in assessing the random effect of clusters. PMID- 15814172 TI - Current and lifetime exposure to working conditions. Do they explain educational differences in subjective health? AB - Previous research has shown that the association between education and health is partly mediated by working conditions. So far, most studies fail to take into account working careers and instead focus on working conditions at one point in time. This study examines the extent to which current and lifetime exposure to working conditions differ between educational groups, and whether taking into account lifetime exposure rather than current exposure improves our understanding of educational differences in health. A representative sample of the Dutch population (n = 1561) with retrospective information about working careers shows that lower educated men are significantly more exposed to adverse working conditions than higher educated men. These differences increase over the life course. Among women there are relatively small educational differences in exposure. Lifetime exposure to adverse working conditions explains a significant part (a third) of the health differences between the highest and lowest educated men. Moreover, measurements of lifetime exposure to working conditions offer a better explanation for educational differences in health than measurements of current exposure. Among women, only relative lifetime exposure to working conditions can explain a small part of the educational differences in health, while current and absolute lifetime exposure do not explain these differences. PMID- 15814173 TI - Exploring the generalisability of the association between income inequality and self-assessed health. AB - A growing between- and within-country literature suggests that the association between income inequality and health reflects individual- or area-level characteristics with which income inequality is associated, rather than the effects of income inequality per se. These studies also suggest that the association between income inequality and health is country-specific. Unresolved methodological issues include the geographical level at which to model the effects of income inequality, and the appropriate statistical methods to use. This study compares the results of single-level and multi-level logistic regression models estimating the association between income inequality and self assessed health in local authorities in Scotland. The results suggest that there is a significant positive association between income inequality and health across local authorities in Scotland, even after adjusting for individual-level socio economic status. They also suggest that there is significant local authority level variation in self-assessed health, but this is small compared to the variation at the individual level. Income and other measures of individuals' socio-economic status are more strongly associated with self-assessed health than income inequality. This study provides further evidence that the income inequality:health association is place-specific. It also suggests that methodological choices regarding the ways of estimating the association between self-assessed health, individual-level socio-economic status and area-level income inequality may not make a substantive difference to the results when contextual effects are small. Further work is required to test the sensitivity of these conclusions to alternative levels of geographical aggregation. PMID- 15814174 TI - Adoption of hospital case management: economic and institutional influences. AB - Case management became prevalent in US hospitals in the 1990s and is believed to be beneficial in controlling resource utilization, improving quality of care, reducing variation of care processes and enhancing both patient and staff satisfaction. This research investigates the adoption of case management by US hospitals at three time periods: 1994, 1997, and 2000. We propose that both economic and institutional factors influence the adoption of this management innovation, with economic factors being more influential in early and mid-periods (1994-1997) and institutional factors being more influential in later periods (after 1997). Using American Hospital Association Annual Survey Data and community data from the Area Resources File, we assess the relationship of baseline (1994) hospital and market characteristics to the likelihood of early adoption compared to late adoption, and mid-adoption compared to late adoption. We confirm that both economic and institutional forces influence the likelihood of early and mid-period adoption of case management compared to late adoption. We conclude that institutional influences aimed at achieving or maintaining legitimacy may be as strong a motivator for hospitals to adopt case management as are economic incentives. Implications for practice and further research are discussed. PMID- 15814175 TI - Influence of waiting time on cost-effectiveness. AB - Economic evaluations of health care programs are intended to support policy decisions and therefore should incorporate elements of the health care environment such as waiting lists. We explore possible relationships between waiting time and the cost-effectiveness of health care programs. The impact of waiting on cost-effectiveness is very scenario dependent and may be substantial, especially if health loss while waiting is partly or completely non-reversible. We argue that economic evaluations of health care programs in countries with waiting lists should consider the possible impact of waiting on costs and health effects. PMID- 15814176 TI - Contracts in the real world: case studies from Southern Africa. AB - The desirability of using the private sector to deliver public services is widely debated internationally. Understanding the nature of contracts that initiate and govern such public-private partnerships, and the extent to which they can define the performance of private providers, is key in addressing the questions that underlie this debate. Such understanding has to be gained through better knowledge of all the influences upon contractual relationships. Environmental and institutional factors have been highlighted as one set of influences in need of more attention. This paper presents case studies of three contracts for primary care services in Southern Africa. It reports aspects of the institutional and environmental context in which they operate, and reflects on the nature of publicly financed primary care as a service to be contracted out. An urban-based private sector contract for a sub-set of primary care services was found to operate very differently from rural-based public sector contracts, which attempted to provide broader coverage. The latter contracts were more loosely defined and operated in a more relational manner. Important environmental influences on incomplete contractual relationships explored here are the nature of the market, scope of services, management capacity and involvement of a public purchaser. The paper illustrates some of the practical challenges for low- and middle-income countries in pursuing a policy of contracting with private providers for public primary care services, and particularly highlights the difficulties of deciding how to divide up responsibility between the public and private sectors and yet maintain a comprehensive service delivery system. PMID- 15814177 TI - Aids, race and the limits of science. AB - In the US, HIV is understood as the virus that causes AIDS, the root of a disease syndrome perceived to indicate an immune system that has ceased to function. These understandings reflect the unquestioned hegemony of Euro/American scientific knowledge, a hegemony that precludes alternative interpretations of life and death, health and disease. This paper argues that HIV/AIDS is more complicated than biomedicine allows, and that the "overmedicalization" of treatment and prevention efforts obscures the significant socio-cultural and political-economic realities that shape the global pandemic, including conceptions of race. The paper specifically focuses on the discourses of bio- and socio-pathology that link African diasporic communities around the globe, and which often seamlessly articulate with structural locations, producing a coherent narrative in which social and moral positions justify and substantiate one another. The analysis here occurs on both these levels: the discursive and the structural. If we are to understand the complex relationships that form the AIDS epidemic, the disciplinary lines imagined between scientific paradigms and the clinical focus on the individual body on one hand, and the social sciences and humanities disciplines on the other, must be breached. If what we call AIDS is a socio-cultural and political-economic phenomenon with biological manifestations, then it is essential that the insights of the social sciences and humanities be brought to bear on finding solutions to the epidemic. PMID- 15814178 TI - Racialized bodies, disabling worlds "they [service providers] always saw me as a client, not as a worker". AB - This paper makes a case for a contextualized reading of intersecting constructs of disability, gender and race as they unfold in the everyday lives of immigrant women with disabilities. Taking the vantage point of Mehrun, a Canadian Muslim woman with polio, we show that the disability, gender and race constructs converge in some contexts and remain separate in others. This is an important consideration as it forestalls a situation where immigrant women's activist work is seen to be confined to the discrete sphere of their own communities. Mehrun's story of migration and settlement as well as her "work" on community integration of persons with disabilities (regardless of race or gender) is a plea for the civil rights of disabled people. At the same time, Mehrun's embodied reality as a racialized woman with disabilities suggests the possibility of making her marginality the epicenter of change. PMID- 15814179 TI - The impact of own and spouse's urinary incontinence on depressive symptoms. AB - This study investigated the impact of own and spouse's urinary incontinence on depressive symptoms. Attention was paid to the possibility that gender and caregiving might be important factors in understanding significant effects. We used negative binomial regression to analyze survey data for 9974 middle-aged and older respondents to the Health and Retirement Study in the USA. Results supported the hypothesis that the respondents' own urinary incontinence was associated with depressive symptoms (unadj. IRR = 1.73, 95% CIs = 1.53, 1.95 for men; unadj. IRR = 1.50, 95% CIs = 1.38, 1.63 for women). Controlling sociodemographic and health variables reduced this relationship, but it remained statistically significant for both men and women. Having an incontinent wife put men at greater risk for depressive symptoms (unadj. IRR = 1.13, 95% CIs = 1.02, 1.25), although this relation became nonsignificant with the addition of control variables. No relation between women's depressive symptoms and husbands' (in)continence status was found. Caregiving was not a significant variable in the adjusted analyses, but spouses' depressive symptoms emerged as a significant predictor of the respondents' own depressive symptoms. Health care providers must be sensitive to the emotional impact of urinary incontinence. Our findings also suggest the importance of considering the patient's mental health within a wider context, particularly including the physical and mental health of the patient's spouse. PMID- 15814180 TI - Gender differences in the relation between social support, problems in parent offspring communication, and depression and anxiety. AB - Gender differences in the buffer-effect of social support in the relation between stressful circumstances and the development of depression and anxiety disorders are widely assumed, but few studies address this three-way interaction between gender, stress, and support. Data in the present study came from the baseline assessment of the Adolescents at Risk for Anxiety and Depression (ARIADNE) study in 502 adolescent and young-adult children of 356 parents in the Netherlands with a depression, panic disorder and/or obsessive-compulsive disorder. Results indicate that the daughters benefit more from social support than the sons when problems in parent-offspring communication are high, but that this effect holds only for depression symptoms and particularly in relation to problems in father offspring communication. Social support does not seem to play a role in the development of anxiety. PMID- 15814181 TI - Stereotactic radiosurgery: the patient's experience. AB - Stereotactic radiosurgery (SRS), a highly technical treatment for life threatening brain tumors and vascular malformations, is provided at one site (Dunedin Hospital) for all New Zealand. The full-day procedure begins with the surgical attachment by bone screws of a metal helmet, followed by computerized tomography with or without angiography, and concludes with focused irradiation of the lesion. Previous SRS research has focused on therapeutic indications and outcomes; we report here patients' experiences of this procedure using both qualitative and quantitative methods. Participants (n = 12) uniformly described SRS as unusual and highly significant, a pivotal event shaping the future. For most, the procedure symbolized relief of symptoms, hope, reduction of risk, and recovery. Descriptions of the disruption produced by the onset of symptoms, diagnostic procedures and SRS, followed by a period of uncertain prognosis, also signaled experiences of liminality. Although patients experienced anxiolysis and described status changes following SRS, their lives are not returned to normal; they continue to inhabit a liminal state between health and normality on one hand, illness and disability on the other. Our findings indicate aspects of the experience of SRS for patients and their families that could assist in technically informing and emotionally supporting patients through this unusual treatment. PMID- 15814182 TI - The experiential knowledge of patients: a new resource for biomedical research? AB - Both governments and patients' movements are increasingly making a plea in favour of the active participation of patients in biomedical research processes. One of the arguments concerns the contribution that patients could make to the relevance and quality of biomedical research based on their 'experiential knowledge'. This article reflects on the validity of patients' experiential knowledge in the context of biomedical research processes. Since a conclusive argument on the validity of patients' experiential knowledge could not be reached on the basis of theoretical reflection alone, a pragmatic approach was chosen that assessed the validity of patients' experiential knowledge in terms of its practical usefulness for biomedical research. Examples of patient participation in biomedical research were sought through literature research and more than 60 interviews with (bio)medical scientists, patients, representatives from patients' organisations, and health professionals in the Netherlands and the United Kingdom. These examples were analysed for a concrete contribution by patients to the research process. Twenty-one cases of patient participation in biomedical research were identified. After further analysis, concrete use of patients' experiential knowledge could be traced for nine of these cases. These findings suggest that patients' experiential knowledge, when translated into explicit demands, ideas, or judgements, can contribute to the relevance and quality of biomedical research. However, its deliberate use would require a more structural and interactive approach to patient participation. Since the implementation of such an approach could face various obstacles in current biomedical research practices, further research will be needed to investigate its feasibility. PMID- 15814183 TI - Assessing patients' preferences for treatments for angina using a modified repertory grid method. AB - A current popular theme in medicine concerns whether and how patients should be involved in treatment choice. Assuming patient involvement is desirable, how should one go about eliciting preferences? A variety of quantitative and qualitative methods exist that may be used for this purpose, one of which is the repertory grid method. This method involves eliciting constructs (reasons) for preferences through comparing sets of three options. This method allows the structured elicitation of the reasons behind individual preferences, but also, when used with generalised procrustes analysis (GPA), allows aggregation of individual data to reveal general preference patterns. In this study the repertory grid method was used to examine patient preferences for angina treatments with the goal of, first, gaining some understanding of general patterns of patient preference, and second, examining the likely utility of the technique in this setting. A sample of 21 patients with mild and stable angina from two general practices in Norfolk, UK was interviewed using the repertory grid method to elicit the constructs underlying their preferences amongst seven angina treatments (including 'no treatment'). Individualised questionnaires were then produced and sent to the patients for self-completion, which required rating the extent to which each construct was relevant for each treatment (scored on visual analogue rating scales). Analysis of the ratings, using GPA, showed that the constructs clustered around two dimensions: 'some treatment' versus 'no treatment', and drug treatment versus surgical treatment. While some treatment was generally preferred to no treatment, individuals varied in preference for drug treatments or surgical treatments. Although the latter were generally perceived as 'effective' they were also perceived, for example, as 'invasive', 'frightening', related to 'negative experiences', and being more appropriate for when symptoms are severe ('proportionate'). We consider the implications of these results for involving patients in choosing amongst treatments. PMID- 15814184 TI - The place of death of cancer patients: can qualitative data add to known factors? AB - Research on the distribution of cancer deaths by setting-hospital, hospice, home, other--is longstanding, but has been given fresh impetus in the UK by policy commitments to increase the proportion of deaths occurring in patients' homes. Studies of factors associated with the location of cancer deaths fall into two main categories: geo-epidemiological interrogations of routinely collected death registration data, and prospective and retrospective cohort studies of terminally ill cancer patients. This paper summarises the findings of these studies and considers the place of death factors that are generated in semi-structured interviews with 15 palliative care service providers working in the Morecambe Bay area of north-west England. These qualitative data are found not only to confirm and considerably enrich understanding of known factors, but also to bring new factors into view. New factors can be grouped under the headings: service infrastructure, patient and carer attitudes, and cultures of practice. Such an approach provides useful information for policy makers and practitioners in palliative care. PMID- 15814185 TI - French hormones: progestins and therapeutic variation in France. AB - Western medicine is seen as universally valid, but in reality it displays a wide range of national and local variability. Our paper focuses on one such case of local variation: the widespread use of progestins in France to treat various pre menopausal conditions as well as for contraception. The case of progestins allows us to explore how specific styles of research may come to dominate a particular local medical culture, and how they are influenced by changing criteria of scientific validity and wider social relations. We argue that in the 1980s and 1990s a single prestigious research-oriented Parisian hospital service played a dominant role in the transformation of progestins into scientifically validated medical practice. This status was not called seriously into question until recently when foreign research on a different form of hormone therapy suggested that risk was associated with their use. We also propose that both the research around and medical use of progestins in France was shaped by the positive attitude of many French women, including feminists, to hormonal therapies and to the non-surgical specialty most closely associated with hormones, medical gynaecology. PMID- 15814186 TI - Do panel surveys make people sick? US arthritis trends in the Health and Retirement Study. AB - Researchers have long viewed large, longitudinal studies as essential for understanding chronic illness and generally superior to cross-sectional studies. In this study, we show that (1) age-specific arthritis prevalence in the longitudinal Health and Retirement Study (HRS) from the United States has risen sharply since its inception in 1992, and (2) this rise is almost surely spurious. In periods for which the data sets are comparable, we find no such increase in the cross-sectional National Health Interview Survey (NHIS), the primary source for prevalence data of chronic conditions in the US. More important, the upward trend in the HRS is not internally consistent: even though prevalence in the HRS rises sharply between 1992 and 1996 for 55-56 year-olds, the prevalence for that age group plummets to its 1992 level among the new cohort added in 1998 and then rises rapidly again between 1998 and 2002. We discuss possible reasons for these discrepancies and demonstrate that they are not due to sample attrition in the HRS. PMID- 15814187 TI - Disease metaphors in new epidemics: the UK media framing of the 2003 SARS epidemic. AB - Since the emergence of HIV/AIDS in the 1980s, social scientists and sociologists of health and illness have been exploring the metaphorical framing of this infectious disease in its social context. Many have focused on the militaristic language used to report and explain this illness, a type of language that has permeated discourses of immunology, bacteriology and infection for at least a century. In this article, we examine how language and metaphor were used in the UK media's coverage of another previously unknown and severe infectious disease: Severe Acute Respiratory Syndrome (SARS). SARS offers an opportunity to explore the cultural framing of a less extraordinary epidemic disease. It therefore provides an analytical counter-weight to the very extensive body of interpretation that has developed around HIV/AIDS. By analysing the total reporting on SARS of five major national newspapers during the epidemic of spring 2003, we investigate how the reporting of SARS in the UK press was framed, and how this related to media, public and governmental responses to the disease. We found that, surprisingly, militaristic language was largely absent, as was the judgemental discourse of plague. Rather, the main conceptual metaphor used was SARS as a killer. SARS as a killer was a single unified entity, not an army or force. We provide some tentative explanations for this shift in linguistic framing by relating it to local political concerns, media cultures, and spatial factors. PMID- 15814188 TI - Do higher status and more autonomous women have longer birth intervals? Results from Cebu, Philippines. AB - We look at whether women's status and autonomy affect birth-to-conception intervals using data from the Cebu Longitudinal Health and Nutrition Survey (CLHNS) in the Philippines. We followed 1123 married, fecund women, aged 25-49, for up to 5 years. In a 1994-1995 survey, women were asked about the timing of their last birth. In 1998-2000, women were asked about any pregnancies since the 1994-1995 survey. Using these two surveys, we calculated birth to conception intervals. Women were censored if they reached their 50th birthday during follow up. We measure autonomy based on whether the wife has the final say in 10 household decisions as measured in the 1994-1995 survey. Using Cox proportional hazards models we find that women with more decision-making autonomy have significantly longer birth-to-conception intervals in unadjusted models. After adjustment for age, wealth, education, other socio-economic variables, and women's status, decision-making autonomy remained a significant predictor in all models. This effect remains even after adjusting for contraceptive use, implying that autonomy influences birth-to-conception intervals through other mechanisms above and beyond increased contraceptive use. Additionally, few of the women's status variables were significantly associated with time to next conception. Women who had their first birth later in life were more likely to conceive during the observation period suggesting that they may be having shorter birth intervals in order to "catch up" with their peers. Maternal and child health-care efforts can help women achieve their desired spacing goals by supporting women's autonomy in addition to ensuring they have accurate information and a range of contraceptive options. PMID- 15814189 TI - D-serine enhances impaired long-term potentiation in CA1 subfield of hippocampal slices from aged senescence-accelerated mouse prone/8. AB - The molecular and cellular mechanisms underlying the cognitive deficiency of senescence-accelerated mouse prone/8 (SAMP8) have been attributed to many pathological changes in neurons. Recently, increasing evidence has shown that astrocytes, by mean of d-serine, involve in the process of synaptic transmission. Here we reported that the long-term potentiation (LTP) in CA1 area of hippocampal slices prepared from 2-, 6- and 12-month-old SAMP8 significantly decreased with age. Meanwhile, the LTP in the slices of 6- and 12-month-old mice markedly decreased below that of the age-matched normal strain SAMR1. Supplement with exogenous d-serine, a main product of astrocytes and a coagonist at the "glycin binding" site of N-methyl-d-aspartate (NMDA) receptors, not only directly enhanced the deficient LTP but also rescued the abolished LTP by d-amino acid oxidase (DAAO) in slices from 12-month-old SAMP8. This ameliorative effect of d serine was inhibited by either AP-V or 5,7-dichlorokynurenic acid (DCKA). These results suggest that absence of d-serine or dysfunction of the astrocytes possibly was one of mechanisms underlying the decrease of NMDA receptor-dependent LTP and cognition in aged SAMP8. PMID- 15814190 TI - The A140V mutation in the MECP2 gene is not a common etiological factor among Brazilian mentally retarded males. AB - In mammals, methyl-CpG binding proteins play a significant role in the control of gene expression through their association with chromatin-remodeling complexes. Mutations in the gene coding for methyl-CpG-binding protein 2 (MECP2) cause Rett syndrome and have also been reported in a number of X-linked mental retardation diseases. In this study, DNA samples from 363 male individuals with syndromic and non-syndromic mental retardation and other psychiatric diseases were screened for A140V (419C>T) mutation in the MECP2 gene, considered the most frequent MECP2 mutation in males. No 419C>T was found suggesting that the A140V mutation in the MECP2 gene is not a common cause of mental retardation in males. Recently, a new and abundant isoform of MECP2 was described, which has an alternative N-terminus, translated from exon 1, that was previously thought to be non-coding and has been excluded from many mutational screening, as well, the 5' and 3' UTR regions. We consider essential proceeding further screening in the whole extension of the MECP2 gene using clinically well-documented and larger sized sample to assure the overall contribution of MECP2 to mental retardation. PMID- 15814191 TI - Age of acquisition modulates the amplitude of the P300 component in spoken word recognition. AB - Words acquired earlier in life are easier to process in adulthood than words acquired later; this is known as the age of acquisition (AoA) effect. The goal of this study was to establish whether the P300 component of event-related potentials (ERPs) is sensitive to AoA. Early-acquired words (12.5%), late acquired words (12.5%) and pseudo-words (75%) were presented in an auditory lexical decision task. The two sets of words were matched for length, word type, concreteness, imageability and, crucially, word frequency. Early-acquired words were recognised faster and more accurately than late-acquired words. In addition, AoA modulated ERP activity in centroparietal electrode sites, with early-acquired words eliciting a larger positivity (P300) than late-acquired words. This is the first study to demonstrate an ERP correlate of AoA effects. An important implication of our findings is that AoA may need to be controlled in ERP studies of lexical processing, especially in designs in which it is likely to be a confound (e.g., studies of lexical category effects). PMID- 15814192 TI - Effects of immersion in virtual reality on postural control. AB - In the present study, we examined the effects of the time lag between visual scene and the head movement in the virtual reality (VR) world on motion sickness and postural control in healthy volunteers. After immersion in VR with additional time lags (from 0 to 0.8 s) to the inherent delay (about 250 ms), the visual vestibular conflict induced a slight motion sickness in experimental subjects, but no change was noticed in the body sway path with eyes open and closed. However, Romberg ratio of body sway path with eyes closed divided by that with eyes open after immersion in VR was significantly decreased in comparison with that before immersion in VR. Since Romberg ratio is an index of visual dependency on postural control, this finding indicates that the immersion in VR decreases the visual dependency on postural control. It is suggested that adaptation to visual-vestibular conflict in VR immersion increases the contribution of vestibular and somatosensory inputs to postural control by ignoring the conflicting delayed visual input in the VR world. VR may be a promising treatment for visual vertigo in vestibular patients with unsuccessful compensation by its ability to induce vestibular and somatosensory reweighing for postural control. PMID- 15814193 TI - Adenosine A1 receptors modulate the discharge activities of inspiratory and biphasic expiratory neurons in the medial region of Nucleus Retrofacialis of neonatal rat in vitro. AB - This study investigated whether adenosine A1 receptors could modulate respiratory rhythm in mammals. Experiments were performed in in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of Nucleus Retrofacialis (mNRF) with the hypoglossal nerve (XII nerve) rootlets retained. The rhythmical discharges of the biphasic expiratory (biphasic E) neurons/inspiratory neurons (I neurons) and activities of the XII nerve rootlets were simultaneously recorded by using extracellular microelectrodes and suction electrodes, respectively. Roles of adenosine A1 receptors in modulation of respiratory rhythm were investigated by administration of the adenosine A1 receptor agonist R-phenylisopropyl-adenosine (R-PIA, 10 microM) and its specific antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 2 microM). DPCPX decreased the respiratory period (27.19%) and expiratory duration (28.27%) of biphasic E neurons and at the same time increased the peak discharge frequency (48.13%). By contrast, R-PIA produced opposite effects. On the other hand, the effects of DPCPX and R-PIA on the I neurons were similar to that on the biphasic E neurons except that R-PIA shortened the discharge duration of I neurons (34.12%) and decreased the peak discharge frequency (37.75%) in the middle phase of inspiration, but not in the initial and terminal phases. These results suggest that adenosine A1 receptors are involved in the phase-switching between expiration and inspiration by affecting biphasic E neurons. Activation of adenosine A1 receptors may modulate the inhibitory synaptic inputs from I neurons to biphasic E neurons. PMID- 15814194 TI - A family-based association study of schizophrenia with polymorphisms at three candidate genes. AB - Clinical researches have shown that there is a genetic contribution to the pathogenesis of schizophrenia. Recent studies have suggested that three genes neuropeptide Y (NPY), phosphoinositide-3-kinase class 3 (PIK3C3) and 14-3-3 eta chain gene (YWHAH) are probably associated with schizophrenia. To replicate these findings, we carried out a family-based study on a sample of 235 trios. Our results suggest that the polymorphisms at the NPY and YWHAH genes are unlikely to be linked with genetic susceptibility to schizophrenia. However, we found significant evidence of preferential transmission of the -432C allele of the PIK3C3 gene in the entire trios (Z=2.91, d.f.=1, P=0.0036) and the male probands trios (Z=2.66, d.f.=1, P=0.0079). PMID- 15814195 TI - Amygdalic levels of dopamine and serotonin rise upon exposure to conditioned fear stress without elevation of glutamate. AB - Conditioned fear is an artificial stress, induced by a stimulus, such as a tone, that does not elicit fear in nature. This fear response is acquired by experimental animals when tone is combined with an unconditioned stimulus, such as electrical foot shock. The amygdala is considered to be the area involved in acquisition, consolidation and recall of fear. A series of previous pharmacological studies showed antagonists of dopamine D1 and D2, glutamate N methyl-D-asparatate and (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors to prevent the acquisition of conditioned fear. However, little is known about the types of neurotransmitters released when conditioned fear is acquired and recalled. The present study was designed to continuously monitor changes in extracellular levels of glutamate, dopamine and serotonin in the amygdala, at the acquisition of conditioned fear on Day 1 and at fear recall in response to a tone as a conditioned stimulus on Day 2, using the in vivo microdialysis method. Glutamate was elevated only on Day 1, while dopamine and serotonin rose on both days. The periods of elevated dopamine and serotonin were longer on Day 1 than on Day 2. These results suggest that greater amounts of glutamate, dopamine and serotonin are necessary for acquisition than for recall of conditioned fear. PMID- 15814196 TI - Protection by dietary zinc in ALS mutant G93A SOD transgenic mice. AB - Mutations to the copper, zinc superoxide dismutase (SOD) gene are responsible for 2-3% of amyotrophic lateral sclerosis (ALS) cases. These mutations result in the protein having a reduced affinity for zinc. SOD becomes toxic to motor neurons when zinc is missing from its active site. Recently, high dosages of zinc (75 and 375 mg/kg/day) have been paradoxically reported to increase the death of G93A mutant SOD transgenic mice [G.J. Groeneveld, J. de Leeuw van Weenen, F.L. van Muiswinkel, H. Veldman, J.H. Veldink, J.H. Wokke, P.R. Bar, L.H. van den Berg, Zinc amplifies mSOD1-mediated toxicity in a transgenic mouse model of amyotrophic lateral sclerosis, Neurosci. Lett. 352 (2003) 175-178]. In contrast, we have found that moderate supplementation of zinc (approximately 12 mg/kg/day) delayed death in G93A-mutant SOD mice by 11 days compared to mice on a zinc-deficient diet. Supplementing zinc with even 18 mg/kg/day resulted in a more rapid death of some mice, consistent with the results of Groenevelt et al. However, large amounts of zinc competitively inhibit copper absorption, which inhibits the copper-dependent ceruloplasmin, and can cause a lethal anemia. We found that supplementing the 18 mg/kg/day dosage of zinc with 0.3 mg/kg/day of copper prevented the early death from zinc treatment alone. These data support a role for moderate levels of dietary zinc potentially protecting against the toxicity of ALS-associated SOD and the protection does not result from depleting copper. PMID- 15814197 TI - Inhibitory effect of CGRP on osteoclast formation by mouse bone marrow cells treated with isoproterenol. AB - The present study was designed to elucidate the mode of action of isoproterenol (Isp; adrenergic beta-agonist) and to characterize the effect of the calcitonin gene-related peptide (CGRP; sensory neuropeptide) on osteoclast formation induced by Isp in a mouse bone marrow culture system. Treatment of mouse bone marrow cells with Isp generated tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cells (MNCs) capable of excavating resorptive pits on dentine slices, and caused an increase in receptor activator of NF-kappaB ligand (RANKL) and a decrease in osteoprotegerin (OPG) production by the marrow cells. The osteoclast formation was significantly inhibited by OPG, suggesting the involvement of the RANKL-RANK system. CGRP inhibited the osteoclast formation caused by Isp or soluble RANKL (s-RANKL) but had no influence on RANKL or OPG production by the bone marrow cells treated with Isp, suggesting that CGRP inhibited the osteoclast formation by interfering with the action of RANKL produced by the Isp-treated bone marrow cells without affecting RANKL or OPG production. This in vitro data suggest the physiological interaction of sympathetic and sensory nerves in osteoclastogenesis in vivo. PMID- 15814198 TI - Spine-type densities of hippocampal CA1 neurons vary in proestrus and estrus rats. AB - Spines-mediated synaptic activity has been associated to learning ability. Dendritic spines from hippocampal CA1 pyramidal neurons of proestrus rats have been reported to be more numerous than in estrus animals, but some behavioral studies have reported a better performance during the estrus stage of the estrous cycle. Because spine shape has been shown to be strongly related to the post synaptic processing of information, a quantitative morphological study related to the proportional density of each type of spine, was conducted in Golgi material of hippocampal CA1 pyramidal cells of proestrus and estrus rats. After three regular estrous cycles had been asserted, seven Sprague-Dawley female adult rats in proestrus and eight in estrus, were used. Mushroom-shaped spines from hippocampal pyramidal cells predominated in proestrous rats, being 15.1% more numerous in this stage than in estrus; while thin spines were the predominant type of spine in estrous animals, being 15.5% more abundant in estrus than in proestrus. The predominance of the mushroom-shaped or thin spines in the hippocampal CA1 pyramidal neurons during the respective stages of the rat estrous cycle, could be related to the organization of the hippocampal activity-dependent mnemonic information. PMID- 15814199 TI - Suppression of Pyk2 attenuated the increased tyrosine phosphorylation of NMDA receptor subunit 2A after brain ischemia in rat hippocampus. AB - Effects of suppressing the protein expression of Pyk2 on increased tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor subunit 2A (NR2A) after brain ischemia in rat hippocampus were studied with immunoprecipitation and immunoblot. Transient (15 min) brain ischemia and reperfusion (I/R) was induced by four-vessel occlusion in Sprague-Dawley (SD) rats. I/R led to increases of tyrosine phosphorylation of NR2A and interaction of Pyk2 and Src kinase with NR2A after 6 h of reperfusion. The increases were attenuated by Pyk2 antisense oligonucleotides intracerebroventricularly infused every 24 h for 4 days before ischemia, but not missense oligonucleotides or vehicle. The antisense also inhibited the increased auto-phosphorylation of Pyk2 and Src kinase, while the protein expression of NR2A or Src kinase had no obvious change under the above conditions. The data suggested that Pyk2 may be involved in facilitating NR2A tyrosine phosphorylation by Src kinase after I/R. PMID- 15814200 TI - PD-168077, a selective dopamine D4 receptor agonist, induces penile erection when injected into the paraventricular nucleus of male rats. AB - The effect of PD-168077 (N-methyl-4-(2-cyanophenyl)piperazynil-3-methylbenzamide maleate), a selective D4 dopamine receptor agonist, injected into the paraventricular nucleus of the hypothalamus on penile erection was studied in male rats. PD-168077 (1-200 ng) induced penile erection in a dose-dependent manner. The minimal effective dose was 50 ng, while the maximal response was found with 200 ng of the compound, which increased penile erection episodes from 0.3+/-0.03 to 1.7+/-0.21. The proerectile effect of PD-168077 was reduced almost completely by L-745,870 (3-(4-[chlorophenyl]piperazin-1-yl)-methyl-1H-pyrrolo[2,3 B]pyridine trihydrochloride), a selective D4 dopamine receptor antagonist, (1 microg) given into the paraventricular nucleus before the D4 dopamine agonist, and by other nonselective dopamine receptor antagonists, such as haloperidol (1 microg) and clozapine (1 microg), which block all dopamine receptor subtypes. The pro-erectile effect of PD-168077 was also reduced by the NO synthase inhibitor NG nitro-L-arginine methylester (25 microg), but not by the oxytocin receptor antagonist d(CH2)5Tyr(Me)2-Orn8-vasotocin (1 microg), when given into the paraventricular nucleus. In spite of its inability to prevent the pro-erectile effect of PD-168077 when given in the paraventricular nucleus, d(CH2)5Tyr(Me)2 Orn8-vasotocin (1 microg) reduced almost completely PD-168077-induced penile erection when given into the lateral ventricles. The present results show that D4 dopamine receptors present in the paraventricular nucleus may influence penile erection by modulating the activity of paraventricular oxytocinergic neurons mediating erectile function. PMID- 15814201 TI - Visual capture of apparent limb position influences tactile temporal order judgments. AB - Shore et al. [D.I. Shore, E. Spry, C. Spence, Spatial modulation of tactile temporal order judgments, Perception (submitted for publication)] recently demonstrated that people find it easier to judge which hand is touched first (in a tactile temporal order judgment task) when their hands are placed far apart rather than close together. In the present study, we used a mirror to manipulate the visually perceived distance between participants' hands, while holding the actual (i.e., proprioceptively-specified) distance between them constant. Participants were asked to determine which of two vibrotactile stimuli, one presented to either index finger using the method of constant stimuli, was presented first. Performance was significantly worse (i.e., the JND was larger) when the hands were perceived (due to the mirror reflection) as being close together rather than further apart. These results highlight the critical role that vision plays in influencing the conscious perception of the temporal order of tactile stimuli. PMID- 15814202 TI - Involvement of serotonin in the hypoglycemic response to 2 Hz electroacupuncture of zusanli acupoint (ST36) in rats. AB - In our previous studies, an insulin-dependent hypoglycemic effect produced by electroacupuncture (EA) was shown to be mediated by endogenous opioid peptides (EOP). In the present study, we applied 2 Hz EA to both zusanli acupoints (ST36) in the test group for 30 min, and to a nonacupoint area in the control group for 30 min to compare the acupoint specific character in the hypoglycemic effect of EA. Assays of plasma beta-endorphin and insulin levels were performed by ELISA kits. The insulin-dependent mechanism of the hypoglycemic effect was also investigated in streptozotocin (STZ)-induced diabetic rats. The mediation of EOP and the role of mu-opioid receptor were examined by naloxone and mu-opioid receptor knockout mice (MOR-KOM). The serotonin depletion was carried out by injecting (i.p.) p-chlorophenylalanine (PCPA); two low doses of serotonin were also injected (i.v.) to analyze the direct effect on plasma glucose levels. The hypoglycemic effect of EA was much greater in rats stimulated at ST36 than in rats receiving the same stimulation at the nonacupoint area. The plasma levels of insulin and beta-endorphin were also significantly elevated after stimulation of both zusanli acupoints, but remained unchanged following stimulation at the nonacupoint area. There was no sharp hypoglycemic response to 2 Hz EA at zusanli acupoint of STZ-induced diabetic rats. However, the hypoglycemic effect of this EA was not totally blocked by the sufficient dose of naloxone (1 mg/kg, i.v.). Additionally, 2 Hz EA at ST36 also showed a sharp decrease in plasma glucose levels of MOR-KOM. Pretreatment with PCPA did not reproduce hypoglycemic response to 2 Hz EA in naloxone-treated rats and MOR-KOM mice. Furthermore, injection of serotonin decreased the plasma glucose levels significantly. Therefore, we suggest that serotonin also involved in the hypoglycemic action of 2 Hz EA at both zusanli acupoints of normal rats. PMID- 15814204 TI - Brain injury in preterm infants--what can the obstetrician do? AB - Mothers at increased risk of preterm birth often receive glucocorticoids (GC), antibiotics and tocolytics by the obstetrician but the question is whether such interventions affect the risk of brain injury and neurological outcome. We suggest that one single course of antenatal GC is the most important treatment that can be offered to patients at risk of preterm birth at 24-34 weeks of gestation to prevent brain injury. Betamethasone seems advantageous to dexamethasone and repeated courses of GC should probably be avoided. Antibiotics given to patients with preterm premature rupture of membranes reduce neonatal morbidity and decrease the risk of sonographic cerebral abnormalities even though the effect on long-term neurological outcome is uncertain. From the perspective of the immature CNS, there is no evidence for treatment with tocolytics even though it allows transfer of the patient to a tertiary center and increases the likelihood of administration of a complete course of corticosteroids which may affect outcome. PMID- 15814205 TI - Clinical measures to preserve cerebral integrity in preterm infants. AB - Impaired psychomotor development, often anteceded by major intraventricular hemorrhage or periventricular leukomalacia, constitutes the most important long term morbidity of very preterm infants. We reviewed randomized controlled trials aimed at reducing the incidence of brain damage, as detected by ultrasound, or neurodevelopmental impairment during follow-up of preterm infants. Preliminary reports of reduced rates of intraventricular hemorrhage obtained with administration of fresh frozen plasma, ethamsylate, phenobarbitone, or morphine have not been confirmed in subsequent larger trials. Early administration of indomethacin may reduce intraventricular hemorrhage without affecting long-term outcome. Pancuronium, inositol, and vitamin E decreased intraventricular hemorrhage rates but later psychomotor development was not examined. Thyroxin supplementation failed to improve neurodevelopmental outcome while protein enrichment of formula and individualized developmental care appear to be beneficial. The largest reductions in cerebral palsy and neurodevelopmental impairment were achieved by avoidance of postnatal steroids. This finding emphasizes the need to include these late endpoints in any randomized trial involving preterm infants. PMID- 15814206 TI - Developmental neuropathology of the second half of gestation. AB - In this review we focus primarily on the events taking place in the second half of gestation. At second trimester end, human brain weight gain accelerates rapidly. Germinal matrix attains maximal absolute volume, only to ablate 50% over two gestational weeks. At 10 weeks of gestation interhemispheric, choroidal, and transverse fissures exist. Germinal matrix hemorrhages peak during its devolution and some of these rupture into the lateral ventricle. By 28 weeks homologous primary sulci are present, having appeared in both hemispheres at slightly different gestational ages. Secondary sulcation, during the third trimester, is hemispherically unique. Despite emphasis on neuronal vulnerability, prevalence of lesions in white matter exceeds that of gray matter and, within white matter, diffuse white matter astrocytosis prevalence exceeds that of focal necroses. Gray matter hypotensive lesions most commonly occur in the upper brainstem and thalami followed by convexity borderzone lesions causing sclerotic microgyria. White matter hypoplasia with normal gray matter volume is sometimes associated with hypomyelination. PMID- 15814207 TI - Electroencephalography and brain damage in preterm infants. AB - Electroencephalography (EEG) is a sensitive method for detection of brain injury in preterm infants. Although the acute and chronic EEG changes are mainly non specific regarding type of damage, they correlate with later neurological and cognitive function. In infants developing brain white matter damage, acute EEG findings include depression of background activity and presence of epileptic seizure activity. The chronic EEG changes associated with white matter injury and abnormal neurological development include delayed maturation, and presence of abundant Rolandic sharp waves. Cognitive limitations in preterm infants have been associated with changes in various sleep measures in EEG's recorded at full term. Continuous EEG-monitoring during neonatal intensive care shows that cerebral electrical activity during this vulnerable period can be affected by several extracerebral factors, e.g. cerebral blood flow, acidosis and some commonly used medications. For diagnosis of brain damage in preterm infants with neurophysiological methods, a combination of early continuous EEG monitoring during the initial intensive care period and full EEG, performed at later stages, is probably optimal. PMID- 15814208 TI - Magnetic resonance and ultrasound brain imaging in preterm infants. AB - Cranial ultrasonography has been used to identify brain injury in preterm neonates for more than two decades. More recently, magnetic resonance imaging has been used to evaluate brain development and pathology in these infants. In this review we describe how well findings from these two imaging modalities agree with histology findings and neuro-developmental outcome. In addition, we discuss studies comparing ultrasound and magnetic resonance imaging findings. PMID- 15814209 TI - The Tei index for evaluation of fetal myocardial performance in sick fetuses. AB - OBJECTIVE: The Tei index is a useful, new, noninvasive Doppler index of combined systolic and diastolic function calculated by isovolumic relaxation time plus isovolumic contraction time divided by ejection time. Sick fetuses were evaluated with the Tei index. METHODS: The study group underwent two-dimensional/Doppler echocardiographic measurement of their Tei index and included 10 monochorionic diamniotic (MD) twin pairs with non-twin to twin transfusion syndrome (TTTS), 4 twin pairs with TTTS, 12 fetuses with intrauterine growth retardation (IUGR), 14 fetuses of diabetic mothers, 3 hydrops fetalis fetuses, 8 fetuses of mothers treated with a tocolytic agent, and 40 normal fetuses (control group). RESULTS: The Tei indices in the following groups were significantly higher than the control: recipient fetuses in TTTS, large for gestational age (LGA) fetuses of diabetic mothers, and fetuses with hydrops fetalis. CONCLUSION: The Tei index may be a useful tool for the assessment of fetal cardiac status in a variety of sick fetuses. Recipient fetuses in TTTS, LGA fetuses of diabetic mothers, and hydrops fetalis fetuses may have abnormal myocardial performance. The Tei index readily provides early detection of diminished myocardial function, particularly ventricular dysfunction. PMID- 15814210 TI - Minor neurological dysfunction, cognitive development, and somatic development at the age of 3 to 7 years after dexamethasone treatment in very-low birth-weight infants. AB - The objective of this study was to assess minor neurological dysfunction, cognitive development, and somatic development after dexamethasone therapy in very-low-birthweight infants. Thirty-three children after dexamethasone treatment were matched to 33 children without dexamethasone treatment. Data were assessed at the age of 3-7 years. Dexamethasone was started between the 7th and the 28th day of life over 7 days with a total dose of 2.35 mg/kg/day. Exclusion criteria were asphyxia, malformations, major surgical interventions, small for gestational age, intraventricular haemorrhage grades III and IV, periventricular leukomalacia, and severe psychomotor retardation. Each child was examined by a neuropediatrician for minor neurological dysfunctions and tested by a psychologist for cognitive development with a Kaufman Assessment Battery for Children and a Draw-a-Man Test. There were no differences in demographic data, growth, and socio-economic status between the two groups. Fine motor skills and gross motor function were significantly better in the control group (p<0.01). In the Draw-a-Man Test, the control group showed better results (p<0.001). There were no differences in development of speech, social development, and the Kaufman Assessment Battery for Children. After dexamethasone treatment, children showed a higher rate of minor neurological dysfunctions. Neurological development was affected even without neurological diagnosis. Further long-term follow-up studies will be necessary to fully evaluate the impact of dexamethasone on neurological and cognitive development. PMID- 15814211 TI - Infant sleep at 10 months of age as a window to cognitive development. AB - This study examined the association between sleep-wake patterns and achievements on the Bayley Scales of Infant Development, in a low-risk sample of 10-month olds. The sleep of 50 infants was recorded at home by means of a small activity monitor (actigraph). More motor activity in sleep and more fragmented sleep pattern were moderately associated with lower mental developmental index (MDI) scores. The psychomotor developmental index (PDI) was not associated with the sleep characteristics. The finding that at the end of the first year, sleep quality accounted for a significant, albeit small, percentage of the cognitive achievement variance is in line with a model that links sleep-wake states and behavior regulation during arousal. PMID- 15814212 TI - Prior pain induces heightened motor responses during clustered care in preterm infants in the NICU. AB - BACKGROUND: Acute pain is a significant stressor for preterm infants in neonatal intensive care units (NICU); however, little is known about the effects of acute pain on subsequent motor responses during clusters of tactile handling. AIMS: (1) To compare facial, body and heart rate reactivity in preterm infants at 32 weeks gestational age (GA) during routine care-giving tasks following a rest period (RCC: diapering, measuring abdominal girth and axillary temperature, mouth care) with their responses to Clustered Care following blood collection (PCC). (2) To examine how GA at birth affects patterns of stress and self-regulatory behaviors during RCC and PCC. STUDY DESIGN: Within-group crossover design (random order). SUBJECTS: Preterm infants, N=54 (mean GA at birth 29.3 +/- 2.2 weeks; mean birth weight 1257 +/- 423 g) were assessed at 32 weeks GA in the NICU. OUTCOME MEASURES: The Newborn Developmental Care and Assessment Program (NIDCAP) and Neonatal Facial Coding System (NFCS) were coded from continuous bedside video recordings. Changes in mean heart rate (HR) were computed using custom physiologic software. RESULTS: All infants had heightened facial, body and HR responses when CC followed a painful procedure compared to when they had not been handled prior to CC. Infants born at earlier GA (<30 weeks) had equal numbers of stress cues during RCC and PCC, but dampened self-regulatory behaviors during PCC. CONCLUSION: Prior pain induces heightened biobehavioral reactivity in preterm infants during subsequent tactile procedures. In addition, clustering care is particularly stressful for infants born at earlier GA. PMID- 15814213 TI - Influences of maternal dietary intake and suckling on breast milk lipid and fatty acid composition in low-income women from Brasilia, Brazil. AB - Diet has a strong effect, while suckling stimulus is believed not to influence breast milk fatty acids. The effect of dietary pattern and suckling on the fatty acid composition of Brazilian women's breast milk was studied. A cross-sectional study was conducted with low-income women living in the central region of Brazil, where dietary DHA is not readily available. Fore and hind milk fatty acids were collected from 77 women on day 15+/-1 postpartum, and information on maternal characteristics and dietary habit was taken. The effect of suckling stimulus was measured by the changes between fore and hind milk. The mean body mass index (BMI) of volunteers was 23.7+/-3.2 kg/m(2), and the milk lipid concentration was 4.8+/-1.2 g/dl. A mixture of traditional and western type of dietary habit, high in fat and sugar was observed. The fatty acids of the subject's milk were 41.93+/ 1.42% saturated, 33.31+/-1.67% monounsaturated and 25.03+/-5.23% polyunsaturated (wt/wt). The DHA level in the milk was 0.34+/-0.19%, similar to the values found in milk of many western societies. DHA and 18:3n-3 levels increased from fore to hind milk (p<0.05). Correlation existed mostly between dietary components and milk 16:0 and oleic acid (18:0). An analysis of classes of fatty acids in milk showed oleic acid (18:0) to present a negative correlation with all milk fatty acid classes. The results suggest that breast milk fatty acids of Brazilian women reflect a western maternal dietary pattern and are influenced by the suckling. PMID- 15814214 TI - Beta-carotene concentrations in maternal and cord blood of smokers and non smokers. AB - BACKGROUND: Smoking in pregnancy can cause a reduction in the transport of beta carotene across the placenta, consequently reflecting on the concentrations of this micronutrient in the newborn baby. AIMS: This study determined the concentrations of beta-carotene in maternal blood and umbilical cord blood in a group of smokers and non-smokers. DESIGN AND SUBJECTS: A total of 215 women and respective newborn babies were selected from a hospital in Joinville city, Brazil, between July and November 2002. Women were divided in two groups according to their smoking habits. beta-Carotene concentration was determined by high-performance liquid chromatography (HPLC). The difference between mean concentrations of beta-carotene in maternal blood and cord blood, in smokers and non-smokers, was determined by analysis of variance (ANOVA). The Student's t-test compared mean concentrations of beta-carotene in maternal blood and cord blood, and the mean ingestion of foods rich in beta-carotene by smokers and non-smokers. RESULTS: There was a statistical significant difference between the concentrations of beta-carotene in maternal blood (S=0.271 micromol/L, NS=0.450 micromol/L; P=0.001) and cord blood (S=0.028 micromol/L, NS=0.045 micromol/L; P=0.001) in smokers (S) and non-smokers (NS). The mean concentration of beta carotene was significantly higher in maternal blood (0.398 micromol/L) than in cord blood (0.041 micromol/L) (P<0.001). The intake of foods rich in beta carotene was lower in smokers, than in non-smokers (P=0.04). CONCLUSIONS: The concentrations of beta-carotene in maternal blood and cord blood were influenced by the use of tobacco. PMID- 15814215 TI - Urinary citrate in preterm and term babies. AB - OBJECTIVES: (1) To determine a normal range for urinary citrate for term babies. (2) To compare urinary citrate measured in ex preterm babies at term with this normal range. (3) To evaluate whether urinary citrate was related to presence of nephrocalcinosis (NC) and chronic Lung Disease (CLD) in these ex preterm babies. STUDY DESIGN: Urinary citrate was measured in 38 healthy term babies (mean birth weight 3.52 kg, mean gestation 41 weeks) at a mean postnatal age of 3 days (1-5 days) and in 53 ex preterm babies (<32 weeks gestation at birth) at term. These preterm babies were part of a larger study on NC in which two renal ultrasound scans were performed at 1 month and term. RESULTS: The normal range for urinary citrate in term babies was 0.025-2.97 (mean 1.03) mmol/l and citrate/creatinine ratio 0.0011-0.852 (mean 0.27). In the ex-preterm urinary citrate was not significantly different (mean 1.1 vs. 1.03, p=0.7232) but urine citrate/creatinine ratio was significantly higher (mean 1.27 vs. 0.27, p=0.0005). There was no significant difference in urinary citrate or ratios of citrate/creatinine and calcium/citrate in the 11 (20.7%) with NC or in the 17 (32%) babies with CLD. There was no significant relationship found between duration of TPN and urinary citrate measured at term. CONCLUSION: We have determined a normal range for urinary citrate in healthy term babies in the first week of life. The range was very wide. Ex preterm babies had similar values at term and there was no association between urinary citrate and NC or CLD. PMID- 15814216 TI - The effect of umbilical venous constriction on placental development, cord length and perinatal outcome. AB - BACKGROUND: Umbilical vein constriction at the fetal abdominal inlet is a common finding after week 13, when the period of umbilical herniation is brought to an end. AIMS: To test the hypothesis that a constricting umbilical ring within physiological ranges affects fetal hemodynamics by either pooling blood in the placenta or restricting nutrient transfer to the fetus and thus shift the birthweight/placental weight (BW/PW) ratio. A constriction could also cause pressure changes and elongation of the cord and possibly be a disadvantage during labour. STUDY DESIGN: Cross-sectional. SUBJECTS: 359 Low-risk singleton pregnancies at 13-40 weeks of gestation. OUTCOME MEASURES: Standard deviation score (z-score) and regression analysis were used to determine the effect of umbilical vein constriction (expressed by increased blood velocity) on birthweight/placental weight ratio (BW/PW), cord length, Apgar score and emergency delivery due to fetal distress. RESULTS: Umbilical venous constriction had a mild but significant effect on BW/PW in male (p=0.018) but not in female fetuses. Increased constriction was also associated with increased length of the cord but only in female fetuses (p=0.019). Cord length was positively related to birthweight and placental weight, but an increased length of the cord was also associated with decreasing BW/PW ratio for the male fetuses only (p=0.044). Increasing degree of venous constriction was associated with Apgar score < or =7 at 1 (p=0.009) but not at 5 min after birth and was not associated with emergency delivery. CONCLUSION: Physiological umbilical venous constriction exerts a mild but significant gender-specific hemodynamic impact on intrauterine development. PMID- 15814217 TI - Distribution of sleep and wakefulness EEG patterns in 24-h recordings of preterm and full-term newborns. AB - The aim of this study was to evaluate the organisation of EEG patterns in 24-h recordings of preterm and near-term neonates. In particular, the distribution of the different EEG codes at different postmenstrual ages (PMA) and the variations of sleep-related EEG pattern organisation was studied, during day (8.00 a.m.-8.00 p.m.) and night (8.00 p.m.-8.00 a.m.) time. The age of appearance of different neonatal EEG patterns, previously described in literature for short lasting records, was confirmed in this 24-h study. The medium-voltage continuous EEG pattern (pattern "3") was less represented approaching term age, in coincidence with the appearance of the two low-voltage continuous patterns ("1" and "2"), which are also related to active sleep and wakefulness. Discontinuous pattern ("7") was also less represented with age, but in day-time only. The percentage of time occupied by this pattern, related to quiet sleep, was significantly higher during day-time hours, than at night. PMID- 15814218 TI - Time domain correlation analysis of heart rate variability in preterm neonates. AB - BACKGROUND AND AIM: A fuller understanding of the neural control mechanisms of heart rate during the early stages of human development would be of great value to obstetric and neonatal management. In this paper, we investigate the correlation between heart rate variability (HRV) and other physiological parameters such as blood pressure and respiration in preterm neonates with the aim of developing a numerical model to explain and predict heart rate variability. STUDY DESIGN AND SUBJECTS: All the required data are readily available for premature babies who are routinely monitored while being nursed in intensive care, and we have collected large data sets for a random group of such neonates. For the quantitative analysis of the data, we have developed a time domain correlation method, which has a number of advantages over the more commonly used power spectral analysis. We have been able to study the dynamics of the different frequency components of HRV by this method. RESULTS: Highly correlated behaviour of the different HRV components, previously observed in our work on fetal HRV, is also present in the neonate, with similar characteristic time constants. Furthermore, the correlation of high-frequency (HF) oscillations of HRV with respiration and that of low-frequency (LF) oscillations of HRV with blood pressure are demonstrated on timescales of a single oscillation. In neonates receiving artificial ventilation, the correlation between HRV and respiration depends on the type of ventilation involved and assumes opposite polarities for the two main types of equipment currently in use. CONCLUSION: We demonstrate that it is possible to analyse HRV quantitatively by calculating the relative gains and characteristic time constants for the correlated parameters and components. PMID- 15814219 TI - Plasma amyloid beta protein 1-42 levels in fetuses with Down syndrome. AB - BACKGROUND: The presence of amyloid plaques in the brains of people with Down syndrome is correlated with the severity and the progression of the disease. The core of the plaques is an amyloid beta (A beta) protein. If a relationship between fetal levels and the presence and severity of the disease could be determined, consideration of an early intervention to reduce brain damage can be proposed. AIM: To study plasma amyloid beta 1-42 levels in fetuses with Down syndrome. STUDY DESIGN: Fetal plasma amyloid beta 1-42 levels were measured using a commercially available immunoassay. The sample size was previously calculated to show a difference with an alpha level of 0.05 and a power (1-beta) of 90%. SUBJECTS: Thirteen fetuses with Down syndrome and 17 controls (22.3+/-2.0 and 21.6+/-1.2 weeks of gestation, respectively). OUTCOME MEASURES: Fetal plasma amyloid beta 1-42 levels. RESULTS: There was no significant difference in plasma amyloid beta 1-42 levels between fetuses with Down syndrome and those with a normal karyotype (193.1+/-48.0 vs. 194.6+/-15.6 pg/mL, respectively). CONCLUSIONS: This result does not support the hypothesis that A beta 1-42 may be related to the severity of brain damage in newborns with Down syndrome. The high levels of this peptide in fetuses without Down syndrome favour a physiological role of these peptides during brain development. PMID- 15814220 TI - Pilot study of the systemic effects of three different screening methods used for retinopathy of prematurity. AB - PURPOSE: This pilot study compared the physiological and behavioural changes in premature infants undergoing three different methods of screening for retinopathy of prematurity (ROP). STUDY DESIGN: Prospective randomized cross-over pilot study. SUBJECTS AND METHODS: Fifteen premature infants requiring screening for ROP were recruited, and physiological and behavioural responses produced by three different methods of screening were compared. The screening methods employed a RetCam 120 and an indirect ophthalmoscope with and without an eyelid speculum. Physiological indices (change in pulse, mean blood pressure and oxygen saturation) and facial responses to pain (brow bulge, eye squeeze, nasolabial fold, mouth opening and the presence of cry) were recorded at five points: before, during and immediately after screening and 10 and 30 min after examination. RESULTS: Screening with the RetCam 120 and the indirect ophthalmoscope with a speculum both caused a greater change in pulse and mean blood pressure and an increase in facial responses to pain during and immediately after screening as compared to the indirect ophthalmoscope without the speculum. RetCam 120 screening caused greater desaturation than the other methods. CONCLUSIONS: Although this was a small sample which limits absolute conclusions, the study showed that screening using a RetCam or a speculum and indirect ophthalmoscope caused more stress to the infant, as indicated by physiological and behavioural changes, than simply screening using an indirect ophthalmoscope without a speculum. These effects should be considered when deciding on the appropriate screening method for examining particularly sick infants. PMID- 15814221 TI - Laryngeal oedema in neonatal apnoea and bradycardia syndrome (a pilot study). AB - PURPOSE: Some preterm infants in general good health continue to present recurrent apnoeas, bradycardias and desaturations (ABD) despite usual treatments. These events may lead to transitory brain hypoxia and to further neurological injury. The purpose of this study has been to evaluate the role of laryngeal oedema in this symptomatology and to assess corticoid treatment. METHOD: Twelve preterm babies born at a median age of 28.5 weeks (range: 26-35 weeks) already showed signs of ABD at a median age of life of 28.5 days (range: 9-80 days). Fiberoptic laryngeal endoscopy was performed on these babies at a median postconceptional age of 34 weeks (range: 31-38 weeks) to detect a possible involvement of the larynx in their ABD. RESULTS: Each patient presented a severe laryngeal oedema compatible with potential obstructive breathing. Half of the cohort (n=6) received inhaled corticosteroids initiated with a short oral dexamethasone treatment for 3 to 5 days (group 1). All the babies improved. The other half (n=6) received only an inhaled topic corticosteroid treatment (group 2). Four of the six babies improved and two needed oral dexamethasone. Laryngoscopic endoscopy was carried out after 1 week of treatment. The picture corresponded with clinical improvement. Recurrence of ABD occurred in 3/12 (25%) of the babies after stopping dexamethasone. No immediate side effects of the procedure or the treatment were observed. CONCLUSION: Laryngeal oedema may be a cause of ABD in preterm newborns. It may arise from oesophageal reflux and/or presence of the feeding tube. It can be diagnosed by atraumatic fiberoptic fibroscopy and successfully treated with corticosteroids. PMID- 15814222 TI - Development of the human cuneatus medialis nucleus: a morphometric evaluation. AB - The morphometric features of the development of the human cuneatus medialis nucleus (CMN) were examined using complete serial celloidin sections of 11 brains: nine fetal brains, the brain of a 2-month-old infant and that of a 63 year-old adult. The morphometric analysis revealed at least four stages in the development of the CMN characterized by the maturation of neurons: (1) immature neuronal stage, (2) preparatory stage, (3) accelerated development stage and (4) postnatal stage. The development of the human CMN accelerates after 32 weeks of gestation (WG), and postnatal qualitative maturation also occurs. From the viewpoint of morphometric evaluation, the CMN gradually matures between 18 and 40 WG without a rapid growth phase as far as we have observed. PMID- 15814223 TI - Fetal rate of behavioral inhibition and preference for novelty during infancy. AB - BACKGROUND: Correlations between infant measures of information processing and later IQ have been established, and a few studies have extended this continuum by examining links between various fetal measures and cognitive measures during infancy. AIMS: This study compared fetal rate of behavioral habituation among infants identified as at high or low cognitive risk. STUDY DESIGN: This study was a retrospective examination of differences in fetal rate of behavioral habituation as a function of cognitive risk status at 6.5 and 9 months postpartum, using an independent two-group design. SUBJECTS: Participants were 32 infants who were born between 36 and 42 weeks gestational age at normal birth weight and had normal APGAR scores. OUTCOME MEASURES: The Fagan Test of Infant Intelligence (FTII) was used to obtain a measure of preference for novelty, which served as the basis for identification of infants at high or low risk for cognitive difficulty. These infants had undergone up to three habituation trials using a vibroacoustic stimulus during fetal development. RESULTS: Infants who were classified as being at high cognitive risk at 6 months had required more trials to habituation as fetuses. CONCLUSIONS: Fetal rate of behavioral habituation is associated with information processing during the first 6 months of infancy. We suggest that fetal habituation may be influenced by early cognitive mechanisms and therefore represents an immature but real form of information processing. PMID- 15814224 TI - The influence of the mode of delivery on circulating cytokine concentrations in the perinatal period. AB - BACKGROUND: Cytokines play an important role during labor and full- or preterm delivery. They influence physical immunity of the fetus-neonate and express a leading role in the perinatal period, being present in maternal and fetal tissues. AIM: To investigate whether cytokine concentrations in the mother, fetus and neonate depend on the labor and the mode of the delivery. STUDY DESIGN: Prospective study. SUBJECTS: Seventy-eight healthy, non-smoking parturients (mean age 28+/-4, range 21-39 years, delivering vaginally: n=52 or by elective cesarean section: n=26) and their single, healthy, appropriate for gestational age, full term neonates. OUTCOME MEASURES: We correlated determined circulating levels of IL-2, sIL-2R, IL-4, sIL-4R, IL-6, sIL-6R, IL-1beta, IL-8, IFN-gamma, TNF-alpha, sTNF RI, sTNF RII and RANTES in the mothers before delivery (MS), the fetuses (UC) and the neonates in days 1 (N1) and 4 (N4) of life, with the mode of delivery. RESULTS: sIL-2R in N1 and N4, sIL-4R in MS, IL-6 in MS and UC, IL-1beta in MS, UC and N1, IFN-gamma in MS and UC, TNF-alpha in UC, N1 and N4, sTNF RI in UC were significantly higher in cases of vaginal delivery than in cases of elective cesarean section (p ranging from 0.0005 to 0.05). CONCLUSIONS: Vaginal delivery promotes the production of various cytokines and their receptors, which are implicated in neonatal immunity. PMID- 15814225 TI - A view on the guidelines for management of retinopathy of prematurity. PMID- 15814226 TI - An investigation into the thermal behaviour of a model drug mixture with amorphous trehalose. AB - The thermal and structural properties of amorphous trehalose mixed with a model drug, paracetamol, have been studied with a view to developing understanding of the thermal events undergone by such binary systems. A physical mixture of paracetamol and spray dried trehalose (1:9 weight ratio) was studied using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), hot stage microscopy (HSM), and variable temperature powder X-ray diffraction (VTPXRD). The presence of the drug resulted in a lower temperature recrystallization exotherm for the trehalose compared to the disaccharide alone. Evidence is presented for the trehalose recrystallization being triggered by the melting rather than simply the presence of the paracetamol particles. HSM studies indicated that the trehalose recrystallized around the drug on heating, with the recrystallization temperature again corresponding to the melting of the drug. VTPXRD indicated that the presence of the drug again lowered the recrystallization temperature of the trehalose, although the trehalose anhydrate diffraction peaks were discernible at a lower temperature for both the pure trehalose and the mixed systems than was observed for the DSC studies, suggesting that the association between recrystallization and drug melting was not apparent when using this approach. It is suggested that while the trehalose recrystallization process is not significantly influenced by the presence of the drug when studied over relatively long time periods such as those used for the VTPXRD studies, the process is triggered by the melting of the paracetamol when short experimental times and scanning conditions are used such as those relevant to DSC studies. These data have implications for the quality control of trehalose products using DSC, the characterisation of the physical structure of the binary systems and the prediction of the corresponding physical stability. PMID- 15814227 TI - Absorption-enhancing effect of glycyrrhizin induced in the presence of capric acid. AB - The absorption-enhancing effect of the simultaneous administration of sodium caprate (Cap-Na) and dipotassium glycyrrhizinate (Grz-K) was investigated to clarify an effect of Grz-K. A combination of 0.1% (w/v) Cap-Na and 2% (w/v) Grz-K had a rapid and long-lasting absorption-enhancing activity in Caco-2 cell monolayers under conditions where Cap-Na and Grz-K showed a weak and no activity, respectively. The simultaneous treatment of a Caco-2 cell monolayer with Cap-Na and Grz-K showed no change in intracellular calcium ion level, although a major mechanism of absorption-enhancing effect for Cap-Na was elevation of intracellular calcium ion level. On the other hand, the simultaneous enhancing effect of Cap-Na and Grz-K was inhibited by H7, a PKC inhibitor. Possibly, Grz-K showed an absorption-enhancing effect via PKC cellular signaling pathway after penetration into cell according to increasing membrane permeability by Cap-Na. The absorption of sCT by the rat colon was enhanced by a combination of 0.1% (w/v) Cap-Na and 2% (w/v) Grz-K, and its effect continued even 9h after the onset of the experiment. Furthermore, the simultaneous treatment of 0.1% (w/v) Cap-Na and 2% (w/v) Grz-K showed a negligible histological changes to the colon mucosal membrane and a negligible toxicity on Caco-2 cell monolayer. A combination of Cap Na and Grz-K shows a synergistic absorption-enhancing effect with little mucosal injury, which is applicable to colon-specific delivery. PMID- 15814228 TI - Development of a single dose tetanus toxoid formulation based on polymeric microspheres: a comparative study of poly(D,L-lactic-co-glycolic acid) versus chitosan microspheres. AB - Stable polymeric microspheres capable of controlled release of tetanus toxoid (TT) for periods ranging from days to over months were developed. TT was stabilized, encapsulated in microspheres prepared from poly(D,L)-lactide-co glycolide (PLGA) and chitosan by using protein stabilizer (trehalose) and its immune response was compared. The influence of co-encapsulated protein stabilizer on tetanus toxoid's stability and release from the microspheres was studied. The protein stabilizer (trehalose) prevented structural losses and aggregation of microencapsulated TT. To neutralize the acids liberated by the biodegradable lactic/glycolic acid-based polymer, we also co-incorporated into the polymer an antacid, (Mg(OH)2), which neutralized the acidity during degradation of the polymer and also prevented TT structural losses and aggregation. The in vitro release experiments with PLGA and chitosan microspheres were performed and the release of TT was increased up to 80-90%. The antigen integrity was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by coomassie brilliant blue staining. The SDS-PAGE analysis confirmed that antigen integrity was not affected by the encapsulation procedure. In addition, the immunogenicity of PLGA and chitosan microspheres based single dose vaccine was evaluated in guinea pigs and compared with multiple doses of alum adsorbed TT. Results indicated that a single injection of PLGA and chitosan microspheres containing TT could maintain the antibody response at a level comparable to the booster injections of conventional alum adsorbed vaccines. The both PLGA and chitosan based stable vaccine formulations produced an equal immune response. Hence chitosan can be used to replace the expensive polymer PLGA. This approach should have potential application in the field of vaccine delivery. PMID- 15814229 TI - Effect of hexacosanol on the characteristics of novel sustained-release allopurinol solid lipospheres (SLS): factorial design application and product evaluation. AB - This investigation involved the evaluation of the effect of hexacosanol (HC, ceryl alcohol), a new hydrophobic wax modifier (WM) in comparison with conventional modifiers, on the development of sustained-release allopurinol (AP) solid lipospheres (SLS) intended for use in a suspension formulation and other oral dosage forms. Various beeswax (BW)/WM blends (composition ratio 1:1) were thus used to prepare SLS by a modified oil-in-water emulsion meltable disperse phase (MDP) encapsulation method without using organic solvents and the influence of these blends on the drug encapsulation efficiency (EE), size distribution and the time for 50% of the drug to be released (t50%) was investigated. Results indicated that incorporation of HC in wall matrix of SLS provided the means to enhance the EE of AP and to modulate the rate of drug release into dissolution media (simulated gastric fluid (S.G.F.: pH 1.2) and simulated intestinal fluid (S.I.F.: pH 7.4). The effects of the process variables; HC concentration, dispersant (pluronic F-68: PF-68) concentration and drug:wax ratio were also studied on the properties of AP-loaded SLS by a 2(3) factorial design. The EE values were in the range of 80.8-92.67%. The only significant parameter affecting (P<0.01) the size and size distribution of the SLS formulations was the amount of the PF-68, whereas the factor with the biggest influence (P<0.05) on the drug EE was the initial loading of AP (in terms of the drug:wax ratio). The amount of HC blended with wax and the initial drug loading significantly (P<0.01) affected the t50% values of all of the formulations. The release of AP was more extended (t50% values (S.I.F.; pH 7.4)=9.91-25.36 h, depending on the drug:wax ratio) and surface morphology of SLS was improved with higher HC content (15%, w/w) formulations. The release patterns fitted the Baker-Lonsdale dissolution kinetics for spherical matrices. A significant decrease of plasma uric acid levels (P<0.05) and hepatic impairment in male rats was observed after oral administration of a SLS (mean size: 120 microm) suspensions of the optimum formulation, compared to suspensions of pure AP. PMID- 15814230 TI - Skin targeted DNA vaccine delivery using electroporation in rabbits. I: efficacy. AB - Genetic immunization through skin is highly desirable as skin has plenty of antigen presenting cells (APCs) and is easily accessible. The purpose of this study was to investigate the effects of electroporation pulse amplitude, pulse length and number of pulses on cutaneous plasmid DNA vaccine delivery and immune responses, following intradermal injection in vivo in rabbits. Expression of the delivered plasmid was studied using a reporter plasmid, coding for beta galactosidase. The efficiency of DNA vaccine delivery was investigated using a DNA vaccine against Hepatitis B, coding for Hepatitis B surface antigen (HBsAg). Serum samples and peripheral blood mononuclear cells (PBMC) were analyzed for humoral and cellular immunity, respectively, following immunization. The expression of transgene in the skin was transient and reached its peak in 2 days post-delivery with 200 and 300 V pulses. The expression levels with 200 and 300 V pulses were 48- and 129-fold higher, respectively, compared with the passive on day 2. In situ histochemical staining of skin with X-gal demonstrated the localized expression of beta-galactosidase with electroporation pulses of 200 and 300 V. Electroporation mediated cutaneous DNA vaccine delivery significantly enhanced both humoral and cellular immune responses (p<0.05) to Hepatitis B compared to passive delivery. The present study demonstrates the enhanced DNA vaccine delivery to skin and immune responses by topical electroporation. Hence, electroporation mediated cutaneous DNA vaccine delivery could be developed as a potential alternative for DNA vaccine delivery. PMID- 15814231 TI - Effects of isoflurane on gastrointestinal motility after brief exposure in rats. AB - In pre-clinical studies, investigation of oral formulations often necessitates the use of general anesthesia to facilitate deposition of material directly into the stomach. Since the effectiveness of intestinal drug absorption is dependent on gastric emptying (GE) and intestinal motility, drugs that influence either will also influence drug absorption. This study investigated gastrointestinal motility in rats after brief exposure to Isoflurane (ISO) general anesthesia for orogastric gavage. The use of metochlopramide was also evaluated. Twenty-five fasted rats were induced with brief ISO anesthesia (<6 min). Rats were gavaged a gelatin capsule (8mm (L) x 2.0mm (o.d.)) containing 9 mg of activated charcoal powder (gastrointestinal marker) and rapidly recovered. Gavage was performed using a 15 cm feeding device with a soft hollow tip to hold the capsule. Study included three groups (60 and 120 min recovery, metochlopramide pre-treatment with 60 min recovery) and control. Animals were sacrificed for exposure and examination of the gastrointestinal tract following the allocated recovery period. Gastrointestinal transit of charcoal was reduced approximately 50% 120 min after brief ISO anesthesia. Metochlopramide pre-treatment did not increase gastrointestinal propulsion despite increased GE. These data warrant consideration in intestinal drug absorption studies where ISO is the anesthetic of choice. PMID- 15814232 TI - Aqueous two-phase systems as a formulation concept for spray-dried protein. AB - This study investigates to what extent an aqueous two-phase system (ATPS) can encapsulate and protect the secondary structure of a protein during spray drying. The ATPSs contained polyvinyl alcohol (PVA) and dextran solutions, in different proportions. A model protein, bovine serum albumin (BSA) and, in some experiments, trehalose were added to the ATPS prior to spray drying. Electron spectroscopy for chemical analysis (ESCA), differential scanning calorimetry (DSC), UV spectrophotometry, size exclusion high-performance liquid chromatography (SEC-HPLC) and Fourier transform infrared spectroscopy (FTIR) were used for analysis of solid and reconstituted samples. The anticipated function of the ATPS was to improve the stability of the protein by preventing interactions with the air-liquid interface during drying and by improving the encapsulation of the protein in the dried powder. BSA was found to preferentially partition to the dextran phase and in the absence of PVA, BSA dominated the powder surface. In samples containing PVA, the polymer mainly covered the powder surface, even though the dextran-rich phase was continuous, thus preventing protein surface interactions and providing improved encapsulation. However, PVA was found to cause partial loss of the native structure of BSA although the protein was well encapsulated during spray drying. PMID- 15814233 TI - Optimization and characterization of controlled release multi-particulate beads coated with starch acetate. AB - The objectives of the present study were (1) to model the effects of process and formulation variables on in vitro release profile of a model drug dyphylline from multi-particulate beads coated with starch acetate (SA); (2) to validate the models using R2 and lack of fit values; (3) to optimize the formulation by response surface methodology (RSM); (4) to characterize the optimized product by thermal, X-ray and infrared spectroscopic analyses. Dyphylline loaded inert beads were coated using organic solution of SA with high degree of substitution. A three-factor, three-level Box-Behnken design was used for the optimization procedure with coating weight gain (X1), plasticizer concentration (X2) and curing temperature (X3) as the independent variables. The regression equation generated for Y5 (cumulative percent drug released after 12 h) was Y5 = 89.83 11.98X1 + 2.82X2 - 4.31X1(2) + 1.90X1X2. Optimization was done by maximizing drug release in 12 h and placing constraints at dissolution time points of 0.5, 1, 4 and 8 h. The drug release data of the optimized product were close to that predicted by the model. The models could explain 99% of variability in responses. Thermal, X-ray and infrared analyses suggested absence of any significant interaction of the drug with the excipients used in the formulation. SEM photographs showed the integrity of the coating layer. PMID- 15814234 TI - Injectable biodegradable temperature-responsive PLGA-PEG-PLGA copolymers: synthesis and effect of copolymer composition on the drug release from the copolymer-based hydrogels. AB - Injectable biodegradable temperature-responsive poly(DL-lactide-co-glycolide-b ethylene glycol-b-DL-lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymers with DL-lactide/glycolide molar ratio ranging from 6/1 to 15/l were synthesized from monomers of DL-lactide, glycolide and polyethylene glycol and characterized by 1H NMR. The resulting copolymers are soluble in water to form free flowing fluid at room temperature but become hydrogels at body temperature. The hydrophobicity of the copolymer increased with the increasing of DL lactide/glycolide molar ratio. In vitro dissolution studies with two different hydrophobic drugs (5-fluorouracil and indomethacin) were performed to study the effect of DL-lactide/glycolide molar ratio on drug release and to elucidate drug release mechanism. The release mechanism for hydrophilic 5-fluorouracil was diffusion-controlled, while hydrophobic indomethacin showed an biphasic profile comprising of an initial diffusion-controlled stage followed by the hydrogel erosion-dominated stage. The effect of DL-lactide/glycolide molar ratio on drug release seemed to be dependent on the drug release mechanism. It has less effect on the drug release during the diffusion-controlled stage, but significantly affected drug release during the hydrogel erosion-controlled stage. Compared with ReGel system, the synthesized copolymers showed a higher gelation temperature and longer period of drug release. The copolymers can solubilize the hydrophobic indomethacin and the solubility (13.7 mg/ml) was increased 3425-fold compared to that in water (4 microg/ml, 25 degrees C). Two methods of physical mixing method and solvent evaporation method were used for drug solubilization and the latter method showed higher solubilization efficiency. PMID- 15814235 TI - Characteristics of interpolyelectrolyte complexes of Eudragit E 100 with sodium alginate. AB - With a view to the application in oral drug delivery formulations, the possibility to form interpolyelectrolyte complexes (IPEC) of Eudragit E 100 (EE) with sodium alginate (AL) was investigated, employing turbidimetry, apparent viscosity measurements, FT-IR and elementary analysis. The interaction or binding ratio of a unit molecule of AL with EE was largely affected by the pH value of the media, showing a change from 1.5:1 to 1:1.25 (0.6650 microg/ml). Five plants were of particular interest, associating good antiplasmodial activity and weak cytotoxicity. These five included Nauclea latifolia with known antiplasmodial activity and four, Fagara macrophylla, Funtumia elastica, Phyllanthus muellerianus and Rauvolfia vomitoria, for which the description of antiplasmodial activity is entirely novel. PMID- 15814261 TI - Effect of Commiphora opobalsamum (L.) Engl. (Balessan) on experimental gastric ulcers and secretion in rats. AB - The ulcer protective potential of an ethanol extract of Commiphora opobalsamum (L.) Engl. (Burseraceae) 'Balessan' was assessed against different acute gastric ulcer models in rats induced by necrotizing agents (80% ethanol, 0.2M NaOH and 25% NaCl), hypothermic restraint stress, pyloric ligation (Shay) and indomethacin. Balessan, 250 and 500 mg/kg administered orally (intraperitoneally in Shay rat model) showed a dose-dependent ulcer protective effects in all the above ulcer models. Besides, the extract offered protection against ethanol induced depletion of stomach wall mucus and reduction in nonprotein sulfhydryl (NP-SH) concentration. Ethanol treatment also caused histopathological lesions of the stomach wall. Pretreatment with Balessan extract provided a complete protection of gastric mucosa through supporting both the offensive and defensive factors. Balessan extract was also showed a large margin of safety without any apparent adverse effects in rats. PMID- 15814262 TI - In vitro estrogenic activities of Chinese medicinal plants traditionally used for the management of menopausal symptoms. AB - The estrogenic activity of 70% EtOH extracts of 32 traditional Chinese medicinal plants, selected according to their reported efficacy for the treatment of menopausal symptoms, was assessed using a recombinant yeast system with both a human estrogen receptor expression plasmid and a reporter plasmid. Among them, 11 (34%) species proved to be active. Polygonum cuspidatum had the highest estrogenic relative potency (RP) (3.28 x 10(-3)), followed by Rheumpalmatum (3.85 x 10(-4)), Cassia obtusifolia (3.49 x 10(-4)), Polygonum multiflorum (2.87 x 10( 4)), Epimedium brevicornum (2.30 x 10(-4)), Psoralea corylifolia (1.90 x 10(-4)), Cynomorium songaricum (1.78 x 10(-4)), Belamcanda chinensis (1.26 x 10(-4)), Scutellaria baicalensis (8.77 x 10(-5)), Astragalus membranaceus (8.47 x 10(-5)) and Pueraria lobata (6.17 x 10(-5)). The EC(50) value of 17beta-estradiol used as the positive control was 0.205+/-0.025 ng/ml (RP=100). This study gave support to the reported efficacy of Chinese medicines used for hormone replacement therapy. PMID- 15814264 TI - Medicinal plant wealth of local communities in some villages in Shimoga District of Karnataka, India. AB - An ethnomedicinal survey (1998-2000) was conducted in three villages of Shimoga district of Karnataka, India, using a questionnaire designed by [Sinha, R.K., 1996. Ethnobotany-The Renaissance of Traditional Herbal Medicine. Ina Shree Publishers, Jaipur, India, 242 pp.]. The herbal practitioners in the study area were interviewed and information on medicinal plants, their local names, habitat and their seasonal availability was collected. The survey revealed the utilization of 47 species of plants belonging to 46 genera in 28 families used to treat 9 infectious and 16 non-infectious diseases. Twelve new claims on ethnomedical knowledge were reported and there were formulations that were similar to that described already in the literature. PMID- 15814263 TI - Effect of propolis, some isolated compounds and its source plant on antibody production. AB - Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. Its biological properties and chemical composition may vary according to the geographic location and to the different plant sources. The possible mechanism of action of propolis as well as of its active compounds has been the subject of researchers in recent years. In this work, first we reported the results of our study on the seasonal effect of the immunomodulatory action of propolis on antibody production in bovine serum albumin (BSA)-immunized rats. Then, we compared the effect of Brazilian and Bulgarian propolis, some isolated compounds and Baccharis extract on anti-BSA antibody levels. Based on the results, we conclude that propolis stimulates antibody production, independently of the season and geographic origin. Caffeic acid, quercetin and Baccharis extract had no effect on antibody production, although the importance of isolated compounds is well reported in other biological assays. Propolis action is a consequence of plant-derived products with synergic effects, while isolated compounds or extracts from its plant sources had no effect in this assay. PMID- 15814265 TI - The ameliorative effect of dates (Phoenix dactylifera L.) on ethanol-induced gastric ulcer in rats. AB - The present work aimed at testing, in a rat model of ethanol-induced gastric ulceration, a local folk medicinal claim that dates are beneficial in gastric ulcers in humans. Aqueous and ethanolic undialyzed and dialyzed extracts from date fruit and pits were given orally to rats at a dose of 4 ml/kg for 14 consecutive days. On the last day of treatment, rats were fasted for 24 h, and were then given ethanol, 80% (1 ml/rat) by gastric intubation to induce gastric ulcer. Rats were killed after 1 h of ethanol exposure, and the incidence and severity of the ulceration were estimated, as well as the concentrations of gastrin in plasma, and histamine and mucus in the gastric mucosa. A single group of rats that were fasted for 24 h, was administered orally with lansoprazole (30 mg/kg), and was given 80% ethanol as above, 8 h thereafter, served as a positive control. The results indicated that the aqueous and ethanolic extracts of the date fruit and, to a lesser extent, date pits, were effective in ameliorating the severity of gastric ulceration and mitigating the ethanol-induced increase in histamine and gastrin concentrations, and the decrease in mucin gastric levels. The ethanolic undialyzed extract was more effective than the rest of the other extracts used. It is postulated that the basis of the gastroprotective action of date extracts may be multi-factorial, and may include an anti-oxidant action. PMID- 15814266 TI - Tanshinone inhibits intimal hyperplasia in the ligated carotid artery in mice. AB - Vascular smooth muscle cell (VSMC) proliferation is considered to play a central role in the development of intimal hyperplasia with pathological artery healing. Danshen, the Salvia miltiorrhiza Bge., has long been regarded as an effective traditional Chinese medicine for cardiovascular diseases. In this paper, the effects of tanshinone (TA), the lipid-soluble pharmacological constituents of danshen, on the intima hyperplasia and proliferating state of VSMC were described in a mouse carotid artery injured by complete cessation of blood flow. This study showed that oral administration of TA could significantly decrease the intimal thickening of injured vessels and proliferating cell nuclear antigen (PCNA) positive VSMC in intimal area. These results suggested that the suppressive effects of TA on intimal hyperplasia might partly result from its inhibitory effect against VSMC proliferation. PMID- 15814267 TI - Antiviral activity in vitro of Urtica dioica L., Parietaria diffusa M. et K. and Sambucus nigra L. AB - Parietaria diffusa M. et K., Urtica dioica L. (Urticaceae) and Sambucus nigra L. (Caprifoliaceae) are plants usually used in popular medicine of central Italy for treating numerous diseases, first of all Herpes zoster. Several plant products have been described as potential antiviral agents, with special attention being devoted to those having retroviruses as etiological agents, including acquired immunodeficiency syndrome (AIDS), in which a retrovirus, the designated human immunodeficiency virus HIV, has been clearly identified as the primary cause of this disease. The present study proposes a preliminary screening of the antiviral activity of Parietaria diffusa, Sambucus nigra and Urtica dioica preparation against the feline immunodeficiency virus (FIV) infection. The feline immunodeficiency virus is a widespread lentivirus of domestic cats sharing numerous biological and pathogenic features with the human immunodeficiency virus (HIV). FIV infection in cats has therefore been proposed as an animal model for AIDS studies with respect to pathogenesis, chemotherapy, and vaccine development [Pedersen, N.C., 1993. Feline immunodeficiency virus infection. In: Levy, J.A. (Ed.), The Retroviridae. Plenum Press, New York; Bendinelli, M., Pistello, M., Lombardi, S., Poli, A., Garzelli, C., Matteucci, D., Ceccherini-Nelli, L., Malvaldi, G., Tozzini, F., 1995. Feline immunodeficiency virus: an interesting model for AIDS studies and an important cat pathogen. Clinical Microbiology Revue 8, 87-112; North, T.W., LaCasse, R.A., 1995. Testing anti-HIV drugs in the FIV model. Nature Medicine 1, 410-411; Matteucci, D., Pistello, M., Mazzetti, P., Giannechini, S., Isola, P., Merico, A., Zaccaro, L., Rizzati, A., Bendinelli, M., 2000. AIDS vaccination studies using feline immunodeficiency virus as a model: immunisation with inactivated whole virus suppresses viraemia levels following intravaginal challenge with infected cells but non-following intravenous challenge with cell-free virus. Vaccine 18, 119-130]. Early studies showed that some of them presented antiviral activity against infection of FIV as assayed by syncytia formation using feline kidney Crandell cells (CrFK). PMID- 15814268 TI - In vitro anti-Helicobacter pylori action of 30 Chinese herbal medicines used to treat ulcer diseases. AB - Infection by Helicobacter pylori has been ascertained to be an important etiologic impetus leading usually to chronic active gastritis and gastric ulcer with growing incidences worldwide. Utilizing as the test pathogen a standard and five clinic strains of Helicobacter pylori, the antibacterial action was assessed in vitro with ethanol extracts of 30 Chinese herbal medicines which have been frequently prescribed since ancient times for treating gastritis-like disorders. Among the 30 tested materials, the ethanol extracts of Abrus cantoniensis (Fabaceae), Saussurea lappa (Asteraceae) and Eugenia caryophyllata (Myrtaceae) were strongly inhibitory to all test strains (MICs: approximately 40 microg/ml), and Hippophae rhamnoides (Elaeagnaceae), Fritillaria thunbergii (Liliaceae), Magnolia officinalis and Schisandra chinensis (Magnoliaceae), Corydalis yanhusuo (Papaveraceae), Citrus reticulata (Rutaceae), Bupleurum chinense and Ligusticum chuanxiong (Apiaceae) substantially active with MICs close to 60.0 microg/ml. As to antibacterial actions of the aqueous extracts of the same drugs, those derived from Cassia obtusifolia (Fabaceae), Fritillaria thunbergii and Eugenia caryophyllata were remarkably inhibitory against all the six Helicobacter pylori strains (MICs: approximately 60 microg/ml). The work compared almost quantitatively the magnitude of the anti-Helicobacter pylori actions of the 30 most prescribed gastritis-treating Chinese herbal drugs, and located as well some source plants where potent anti-Helicobacter pylori phytochemicals could be characterized. PMID- 15814269 TI - Effect of an avocado oil-rich diet over an angiotensin II-induced blood pressure response. AB - We studied the effect of an avocado oil-rich diet on (1) the blood pressure response to angiotensin II (AngII) and (2) the fatty acid composition of cardiac and renal membranes on male Wistar rats. The avocado oil-rich diet induced a slightly higher AngII-induced blood pressure response in the rats as compared to the control rats. In cardiac microsomes, avocado oil induced an increase in oleic acid content (13.18+/-0.33% versus 15.46+/-0.59%), while in renal microsomes, the oil decreased alpha-linolenic acid content (0.34+/-0.02% versus 0.16+/-0.12%), but increased the arachidonic acid proportion (24.02+/-0.54% versus 26.25+/ 0.54%), compared to control. In conclusion, avocado oil-rich diet modifies the fatty acid content in cardiac and renal membranes in a tissue-specific manner. The rise in renal arachidonic acid suggests that diet content can be a key factor in vascular responses. PMID- 15814270 TI - Inhibitory effect of jaceosidin isolated from Artemisiaargyi on the function of E6 and E7 oncoproteins of HPV 16. AB - Jaceosidin (4',5,7-trihydroxy-3',6-dimethoxyflavone) was isolated from Artemisia argyi as a putative oncogene inhibitor. Jaceosidin inhibited binding between oncoprotein E6 of the human papillomavirus and the p53 tumor suppressor protein. In addition, jaceosidin inhibited binding between the E7 oncoprotein and the Rb tumor suppressor protein, and also inhibited the function of HPV-16 harboring cervical cancer cells, including SiHa and CaSki. Collectively, jaceosidin inhibited the functions of the E6 and E7 oncoproteins of the human papillomavirus, suggesting that this compound might be used as a potential drug for the treatment of cervical cancers associated with the human papillomavirus. PMID- 15814271 TI - Hypolipidemic activity of aqueous extract of Capparis spinosa L. in normal and diabetic rats. AB - The purpose of this study was to examine the effect of single and repeated oral administrations of the aqueous extract of Capparis spinosa L. (CS) at a dose of 20mg/kg on lipid metabolism in normal and streptozotocin-induced diabetic rats. In normal rats, the aqueous extract of CS induced a significant decrease on plasma triglycerides concentrations 1 week (p<0.05) and 2 weeks (p<0.01) after once daily repeated oral administration. A significant decrease of plasma cholesterol levels was also observed 4 days (p<0.05) and 1 week (p<0.05) after repeated oral administration. In diabetic rats, CS treatment caused a significant decrease of plasma triglycerides levels after repeated oral administration. Four days after repeated oral administration of aqueous CS extract, the plasma cholesterol levels were significantly decreased (p<0.05) and still dropped after 2 weeks (p<0.01). On the other hand, the repeated oral administration of CS aqueous extract caused a significant decrease of body weight 4 days after repeated oral treatment in diabetic rats (p<0.05). We conclude that the aqueous extract of CS (20 mg/kg) exhibits a potent lipid lowering activity in both normal and severe hyperglycemic rats after repeated oral administration of CS aqueous extract. PMID- 15814272 TI - Antimalarial remedies in French Guiana: a knowledge attitudes and practices study. AB - A "knowledge attitudes and practices" study about malaria treatments was undertaken in French Guiana, along with an ethnopharmacological study. One hundred and seventeen people from five different groups and nationalities (Creole, Palikur, Galibi, Brazilian, and European) answered the questionnaire. The results were analysed using univariate and multivariate statistical analysis. First, we evaluated the overall knowledge about malaria from the interviewed people. According to bio-medical concepts, we noticed that they have a good knowledge of this illness. Secondly, we studied the treatment used by sick people during their last malaria attack. We demonstrated that, although bio-medical treatment is available in this area, people use both modern drugs and traditional remedies. Finally, preventive attitudes have been examined. One-third of the interviewed people drink regularly some herbal remedy to prevent febrile illnesses and malaria, thus displaying a strong concern about this disease. The ethnopharmacological study highlighted the frequent use of traditional remedies, along with their mode of preparation and administration. A total of 34 different species (both from flora and fauna) have been registered as antimalarial. Twenty seven are used for curative purposes, 20 as preventive and 13 of them are used for both purposes. Quassia amara (Simaroubaceae) whose antimalarial activity has already been demonstrated was the species most frequently used as antimalarial for curative and preventive purposes. PMID- 15814273 TI - Inhibitory effects of Okbyungpoong-Gamhmi on anaphylactic responses. AB - We investigated the effect of a herbal formulation Okbyungpoong-Gamhmi (OG) on mast cell-dependent anaphylactic reactions by intra-rectal administration. OG concentration dependently inhibited compound 48/80-induced anaphylaxis-like response and ear swelling response with doses of 0.01-1g/kg. OG also inhibited the passive cutaneous anaphylaxis at the same concentrations. The histamine release induced by compound 48/80 or IgE from the rat peritoneal mast cells was reduced by 64.2 and 63.6%, respectively, at 1g/l. These results provide evidence that intra-rectal therapy of OG may be beneficial in the treatment of anaphylactic response. PMID- 15814274 TI - Alkaloids from Boophane disticha with affinity to the serotonin transporter in rat brain. AB - Bulbs and leaves of Boophane disticha are used in South African traditional medicine in the treatment of anxiety. Crude extracts of the leaves have shown affinity to the SSRI site on the serotonin transporter in a radioligand binding assay. In this study, two compounds, buphanadrine and buphanamine, were isolated by bioassay-guided fractionation on VLC and preparative TLC. The structures of the compounds were determined by (1)H and (13)C NMR. Fractions were tested for affinity to the serotonin transporter in a binding assay using [(3)H]-citalopram as ligand. The IC(50) values of buphanidrine and buphanamine were 274 microM (K(i)=132 microM) and 1799 microM (K(i)=868 microM), respectively. The two alkaloids were also tested for affinity to the 5HT(1A) receptor, but only showed slight affinity. PMID- 15814275 TI - Preoperative administration of FTY720 prolonged renal allograft survival. AB - BACKGROUND: FTY729 is an immunomodulator obtained by chemical modification of Myriocin(ISI-1) which exists in the culture filtrate of an ascomycete, Isaria sinclairii. It has been reported that postoperative administration of FTY720 prolonged survival of various kinds of transplanted organs. In the present study, we evaluated the effect of 2-day preoperative administration of FTY 720 on graft survival. MATERIALS AND METHODS: We used a rat renal transplantation model in which Wistar King Aptekman Hokkaido (WKAH, RT1K) served as the organ donor and Lewis (LEW, RTl) as the recipient. FTY720 was given to the recipients consecutively 2 days (day-2, day-1) before transplantation at the doses of 1, 3 or 5 mg/kg/day. Renal allograft survivals, hematological parameters of recipient blood and phenotypic analysis of recipient splenic cells and graft infiltrate were evaluated. RESULTS: Consecutive 2-day preoperative oral administration of FTY 720 at the doses of 1, 3 or 5 mg/kg/day significantly prolonged WKAH allograft survivals compared with those of the untreated recipients. The number of peripheral blood lymphocytes was markedly decreased in the recipients treated with FTY720 at the doses of 3 mg/kg/day or 5 mg/kg/day on the 5th postoperative day. Preoperative FTY 720 administration significantly decreased the number of CD4 positive cells and the percentage of interleukin 2 receptor (IL-2 R) positive cells infiltrating both spleen and allograft at the dose of 3 mg/kg/day or 5 mg/kg/day. CONCLUSION: FTY 720 could act as a safe and potent immunomodulator by decreasing the number of peripheral lymphocytes, especially CD4 positive cells and IL-2R positive cells when it is given to the recipient preoperatively. PMID- 15814276 TI - The bacterial flora of alpha-Gal knockout mice express the alpha-Gal epitope comparable to wild type mice. AB - The human genome possesses pseudogenes for the enzyme alpha1,3 galactosyltransferase and hence, human cells and tissues do not express the Galalpha terminated trisaccharide structure Galalpha1-3Galbeta1-4GlcNAc, the so called alpha-Gal epitope. Circulating antibodies specific for this carbohydrate epitope are, however, present in high amounts in humans. It has previously been hypothesized that the antibody production is induced by the presence of the alpha Gal epitope in the cell walls of the enteric flora, especially Enterobacteriaceae spp. However, in mice, in which the epitope has been deleted by targeted mutation of the gal-transferase gene, alpha-Gal antibodies do not appear without prior immunization, although the mice through their growth probably have been exposed to a normal bacterial flora of e.g. Enterobacteriaceae spp. It is unknown whether there are different types of immune reactions to antigenic carbohydrate expressing bacteria and whether there are discrepancies in the enteric flora between these knockout mice and their wild type litter mates. In this study the enteric flora of alpha-Gal knockout and wild type mice was compared both in relation to the prevalence of different types of bacteria in the two groups of mice, as well as in relation to the expression of the epitope on the surface of Enterobacteriaceae spp. Our results showed that the enteric flora did not differ significantly between knockout and wild type mice and that it was comparable to the flora known to be present in the intestines of other mice. All Enterobacteriaceae spp. examined expressed the alpha-Gal epitope no matter whether they were isolated from knockout or wild type mice. It is, therefore, discussed whether it is more reasonable to assume that alpha-Gal antibodies in mammals that do not produce alpha1,3 galactosyltransferase such as in the knock mice and in humans are the result of another antigen stimulant than these common representatives of the enteric flora, that we isolated from the two types of mice. Possible candidates for a carrier in humans could be bacteria or viruses not isolated from barrier-bred mice. PMID- 15814277 TI - Prolonged survival of donor-specific rat intestinal allograft by administration of bone-marrow-derived immature dendritic cells. AB - It has been reported that intraportal administration of donor antigens induced donor-specific hyporesponsiveness. We studied here the effects of transplantation of BM-derived immature dendritic cells (imDCs) and mature DCs (mDCs) via portal vein on rat small intestinal allograft survival. This study comprised four treatment groups: 1) untreated controls; 2) FK506 alone; 3) intraportal donor specific BM-derived imDCs transplantation+FK506; 4) mDCs/Tx+FK506. Allograft survival was minimal in control group (5.2+/-0.8 days) and maximal in imDC+FK506 group (28.4+/-3.0 days). The rats in mDC+FK506 group showed systemic inflammatory reaction due to GVHR, and died within 10 days after transplantation. The in vitro MLR reaction using imDCs was also strongly inhibited both in direct and indirect recognition pathways. The impact of imDCs for the specific induction of transplant tolerance may suggest that immunization with donor-specific imDCs has therapeutic potential in organ transplantation. PMID- 15814278 TI - Contribution of CD4+ and CD8+ T cells and interferon-gamma to the progress of chronic rejection of kidney allografts: the Th1 response mediates both acute and chronic rejection. AB - T cells mediating chronic rejection (CR) of human kidney allografts were characterized by comparing them with those mediating acute rejection (AR). Two lines of analysis were performed using biopsy specimens (23 CR and 8 AR). First, the extent of infiltration of CD4+ and CD8+ T cells into allografts was assessed from mRNA expression of CD4 and CD8. The group of CR specimens was not significantly different from the group of AR specimens in terms of the extent of CD4+ and CD8+ T cell infiltration, underlining the importance of the immunological contribution to the progress of CR. Second, Th1/Th2 polarization in infiltrating T cells was investigated by measuring mRNA expression of interferon gamma (IFN-gamma; a Th1 cytokine) and interleukin 4 (IL-4; a Th2 cytokine). IFN gamma expression was detected in most CR specimens, and was not significantly different between the group of CR specimens and the group of AR specimens. On the other hand, IL-4 expression was detected in only two CR specimens and one AR specimen; from its pathological features, the AR in this last case was concomitant with CR. These results suggest that most cases of CR and of AR are mediated by Th1 mechanisms, although some cases of CR show features of both Th1 and Th2. PMID- 15814279 TI - Impairment of recipient cytolytic activity attenuates allograft vasculopathy. AB - We investigated the role of CD4+ and CD8+ T subsets as well as T cell cytolytic effector mechanisms in the aortic allograft model of allograft vasculopathy using CD4 and CD8 gene knockout mice (CD4(-/-), CD8(-/-)) and mice deficient in cytolytic effector pathways. Medial apoptosis at 2 weeks was reduced in CD8(-/-) mice and in mice where cytotoxic T cell activity was compromised. At 8 weeks, substantial medial damage was observed in wild-type (WT) and CD4(-/-) recipients but medial preservation was evident in CD8(-/-) mice and in mice with impaired cytotoxic T cell activity. The intima/media ratio, a comprehensive measure of allograft vasculopathy, was similar in WT and CD4(-/-) recipients but was significantly reduced in CD8(-/-) mice and mice with impaired cytotoxic T cell activity. These data indicate that CD8+ T cells contribute to the vascular remodeling that is characteristic of allograft vasculopathy. They also show that CD8+ T cells participate in allograft vasculopathy in the absence of CD4+ T cell help. We further demonstrated that WT mice exhibited robust allograft vasculopathy in the presence of cyclosporin A immunosuppression but that allograft vasculopathy was ablated in cyclosporin-treated CD8(-/-) mice. This supports the hypothesis that non-CD8+ T cell effector mechanisms are sensitive to calcineurin inhibitor therapy but that CD8+ T cell-mediated allograft vasculopathy is refractory to such treatment. Taken together, our data suggest that CD8+ T cells contribute to the induction of vascular remodeling in allograft vasculopathy and provide evidence that novel therapies which target CD8+ T cell effector function might be effective in mitigating AV in the clinical setting. PMID- 15814280 TI - The impact of pharmacogenomic factors on acute persistent rejection in adult lung transplant patients. AB - Persistent rejection in the face of treatment and multiple episodes of rejection are associated with the development of chronic rejection and graft loss in solid organ transplantation. The factors that create an environment for rejection that persists in the face of treatment are as yet not understood. The objective of this study was to evaluate the risk factors, including human multidrug resistance gene (MDR1), cytochrome P4503A5 (CYP3A5) and cytokine gene polymorphisms, associated with acute persistent rejection (APR) in lung transplant patients. One hundred and twenty-five adult lung transplant patients were studied. MDR1 G2677T, C3435T and CYP3A5 polymorphisms were assessed by direct sequencing of the polymorphic region in patient DNA. Cytokine genotyping for five cytokines was performed using the polymerase chain reaction-sequence specific primers (PCR-SSP) technique. Multivariate regression analysis was used to identify the predictors of acute persistent rejection. The dependent variable was the presence or absence of acute persistent rejection based on lung biopsies during the first postoperative year. The independent variables were MDR1 G2677T and C3435T, CYP4503A5 and cytokine polymorphisms, survival status, age, gender, survival days and HLA mismatches. The MDR1 C3435T polymorphism and age were independently associated with acute persistent rejection (p = 0.025, odds ratio = 0.29, 95% CI 0.1-0.86 and p = 0.016, odds ratio = 0.94, 95% CI 0.89-0.98, respectively). For the MDR1 C3435T polymorphism, 72% of patients with the C allele had acute persistent rejection in comparison to 52% for TT patients (p = 0.04). For age, a significant difference was found between the nonrejection group and the rejection group (mean+/-S.D. 52.1+/-11.2 vs. 44.4+/-12.3, p = 0.01). This is the first report of the association of a drug disposition genotype with drug-resistant acute rejection in organ transplant patients. The major predictor of acute persistent rejection in the first postoperative year for lung transplant patients was the MDR1 C3435T genotype. This association could be due to drug resistance, altered drug disposition or other immunologic effects associated with P glycoprotein (P-gp) function. Future prospective treatment algorithms should be developed that will incorporate the knowledge of gene polymorphisms into treatment regimens to improve the outcome following lung transplantation. PMID- 15814281 TI - Daclizumab therapy in kidney transplantation-different mechanisms of action in- versus ex-vivo? AB - Stimulated human T cells from healthy volunteers demonstrate attenuated early interleukin (IL)-2 receptor (R) signaling in the presence of daclizumab (Dac). Aiming to confirm that this ex-vivo effect of Dac is also observed in-vivo, we studied T cells from 3 kidney transplant recipients before and 2-3 weeks and 4-6 months after transplantation. We found by flow cytometry that T cells obtained pre-transplant and stimulated ex-vivo with phytohemeagglutinine upregulated the IL-2R alpha-(CD25) and beta-(CD122) chains as expected. Moreover, exogenous IL-2 induced characteristic tyrosine phosphorylation events detectable by immunoblotting in these cells. However, T cells studied post-transplant neither exhibited CD25 or -122 upregulation nor IL-2-induced tyrosine phosphorylation events, indicating broad, persistent suppression of the IL-2R signaling machinery which thus appears largely inaccessible for Dac in actual transplant recipients. We therefore conclude that the clinical efficacy of this agent may depend on additional mechanisms in-vivo other than those identified ex-vivo. PMID- 15814282 TI - The cytokine gene polymorphisms in patients with chronic kidney graft rejection. AB - Chronic allograft rejection remains an important cause of morbidity after kidney transplantation. The aim of the study was to examine the association between IL 2, IL-6 and TNF-alpha promoter polymorphisms and chronic kidney allograft rejection. The study included 64 patients with long-term stable graft function and 62 with chronic allograft nephropathy. Among patients with chronic allograft nephropathy a statistically significant prevalence of the IL-6 CC genotype associated with low IL-6 expression was observed (p < 0.01, OR 3.18; 95% CI 1.27 8.15). There were no statistically significant differences in distribution of IL 2 and TNF-alpha genotypes between patients with stable graft function and chronic allograft rejection. The results of present study suggest that the genetically determined low IL-6 production may be the risk factor of chronic allograft nephropathy development. PMID- 15814283 TI - Serum analysis after transplant nephrectomy reveals restricted antibody specificity patterns against structurally defined HLA class I mismatches. AB - This study deals with HLA-mismatched kidney transplants that have been removed following rejection. Sera from 27 patients were screened for HLA-specific antibodies by direct complement-dependent lymphocytotoxicity with HLA-typed cell panels. Circulating donor-specific antibodies were detected in 3 cases (11%) before and in 26 cases (97%) after allograft nephrectomy. These findings demonstrate the production of donor-specific antibodies in patients with rejected transplants, but in most cases, they were undetectable before nephrectomy, because the graft had adsorbed them. With an HLAMatchmaker-based serum analysis program, we observed restricted antibody specificity patterns against amino acid triplet-defined epitopes on donor HLA-A,B antigens. Many donor triplets were non reactive while others were apparently recognized by antibodies. In some patients, the donor triplet specific antibodies persisted for a long time whereas in many other patients, they became undetectable after a few months. The characterization of the antibody specificity profiles of post-allograft nephrectomy sera is clinically useful in defining criteria of HLA mismatch acceptability for sensitized patients awaiting another transplant. It provides also opportunities for determining the relative immunogenicity of mismatched triplets. PMID- 15814284 TI - European strategies against the parasite transfusion risk. AB - Protozoal infections are endemic in mainly tropical low income countries, affecting millions of people. Malaria, American trypanosomiasis (Trypanosoma cruzi/Chagas disease) and protozoal tickborne diseases (e.g. Babesia) can be efficiently transmitted by transfusion of cellular blood components. In non endemic areas like Europe malaria, Chagas disease and Babesia are imported diseases resulting of travelling to endemic areas and migration of autochthons from these endemic areas. A recent International Forum showed that in Europe, as well as the USA, prevention of transfusion-associated protozoal infections depend mainly on selection of donors using questionnaires. Most countries divide donors at risk for malaria in two groups: individuals who have lived in the first 5 years of their life in malaria endemic areas and those who are borne and residing in non-endemic areas and visited the endemic area(s). The first category of donors is rejected for 3 years after their last visit to the endemic area, and in one country such donors are permanently rejected. In some countries such donors are accepted after 4 months-3 years, provided a test for malaria is non-reactive. Persons from non-endemic areas, who visited the malaria endemic area, are rejected for 4-12 months. Some countries reject these donors for 3 years or permanently when they resided for more than 6 months in the endemic area. The rejection rate of donors for malaria risk in the various countries was 0.003 0.43% of all donations. Over the last decade only a few cases of TT-malaria were reported in the various countries. In several countries donors are questioned for risk of T. cruzi infection. In some countries donors are excluded when they (or their mothers) were born in South or Central America, if they received a blood transfusion in these areas and if they lived in rural areas in these endemic countries for more than 4 weeks. In none of the countries donors are asked if they had Babesia or Leishmania. At present implemented measures to prevent TT malaria in the European countries are probably highly effective. More research is needed to establish the theoretical risk of TT-T. cruzi and TT-Leishmania infection in Europe, before preventive measures may be considered. PMID- 15814285 TI - Hypoxia and anemia: effects on tumor biology and treatment resistance. AB - In locally advanced solid tumors, oxygen (O2) delivery is frequently reduced or even abolished. This is due to abnormalities of the tumor microvasculature, adverse diffusion geometries, and tumor-associated and/or therapy-induced anemia. Up to 50-60% of locally advanced solid tumors may exhibit hypoxic and/or anoxic tissue areas that are heterogeneously distributed within the tumor mass. In approximately 30% of pretreatment patients, a decreased O2 transport capacity of the blood as a result of tumor-associated anemia can greatly contribute to the development of tumor hypoxia. While normal tissues can compensate for this O2 deficiency status by a rise in blood flow rate, locally advanced tumors (or at least larger tumor areas) cannot adequately counteract the restriction in O2 supply and thus the development of hypoxia. Hypoxia-induced alteration in gene expression and thus in the proteome (< 1% O2, or < 7 mmHg), and/or genome changes (< 0.1% O2, or < 0.7 mmHg) may promote tumor progression via mechanisms enabling cells to overcome nutritive deprivation, to escape from the hostile metabolic microenvironment and to favor unrestricted growth. Sustained hypoxia may thus lead to cellular changes resulting in a more clinically aggressive phenotype. In addition, hypoxia is known to directly or indirectly confer resistance to X- and gamma-radiation, and some chemotherapies leading to treatment failures. Whereas strong evidence has accumulated that hypoxia plays a pivotal role in tumor progression and acquired treatment resistance, the mechanism(s) by which treatment efficacy and survival may be compromised by anemia (independent of hypoxia) are not fully understood. PMID- 15814286 TI - [West Nile virus (WNV): generalities and implications for blood transfusion]. AB - West Nile virus (WNV) is an arbovirus (genus Flavivirus, Family Flaviviridae, transmitted to humans by mosquito bite. In most cases (80%), human infection remains asymptomatic. Severe central nervous system complications (encephalitis and meningoencephalitis) are rare. In the Old World, the virus circulation has been demonstrated in Asia, Australia, Africa, Middle East and Europe. Several outbreaks in humans have been described. Following its introduction into North America in 1999, WN virus has been responsible of a large number of human cases in USA and Canada. For the first time, viral transmission by blood products was clearly demonstrated in USA in 2002. In France, the presence of virus has been reported in the Southeastern departments since 1962. In 2003, the occurrence of humans cases at specific geographical foci urged the French National Blood Agency (etablissement francais du sang) to take preventive measures for evaluating the virus transmission risks. PMID- 15814287 TI - Transposition of the Blood Directive into national law--the perspective of Portugal. AB - European union has published a directive 2002/98/CE in January 2003. The goal of the directive is to standardize the requirements of safety and quality throughout the blood transfusion chain in the countries of the European union. Member states have to implement the principles of this directive in their own regulation. Portugal has already identified what will be the consequences of the new regulation in the health system. PMID- 15814288 TI - Transposing the E. E. U. Blood Directive into national law. Perspective of Slovenia. AB - Transposing the Directive (2002/98/CE) into the national legislation of EU Member States represents the basis for an equal minimal safe blood transfusion and a demanding job for each of these countries. In order to do this Slovenia will need to somewhat change and supplement the legislation adopted in 2000. In Slovenia, the introduction of the quality assurance system brought on the need for changes in the field of transfusion medicine some years ago. The Directive (2002/98/CE) above all signifies obligatory supplements in the field of organization of the transfusion services and especially organization on a national level. In the field of blood donations the development of the profession itself dictates changes. If and when the Directive (2002/98/CE) refers to clinical work, it will be easier to assert the demands for a more rational use of blood, its traceability and follow-ups of adverse events of a blood transfusion. An obligation to take into account the demands and standards of the Directive (2002/98/CE) will accelerate the fulfillment of numerous tasks which we have set for ourselves in the past in order to increase the safety of blood transfusion. PMID- 15814289 TI - [Paramedical staff knowledge and practice related to the blood transfusion safety]. AB - Blood transfusion is a complex activity, involving many actors. It is a high risk activity which couldn't be controlled without the use of specific methods. Health care workers beliefs and organisational factors are two major issues for the blood transfusion safety. Our study objectives were to describe the paramedical staff's knowledge and practice regarding blood transfusion safety and to identify factors that are related to them. We carried out a cross sectional study. The information was gathered by using a questionnaire. The latter was developed by foreign teams and adapted to the local context. Two outcome measures were used: the knowledge and practice score and the proportion of true answers. The study showed that only 15% of the interviewed persons have had a score less than 30 (scale range from 0 to 100), i.e. only 15% have had appropriate knowledge and practice with no negative consequences for the patient safety. 13.8% of the study population provided right answers related to the biologic exams required before red cells transfusion and 34% for the abnormal reaction circumstances. These results underscores the importance of strategies improving the quality and the safety of blood transfusion, i.e. the continuous medical education, implementing a blood transfusion information system and the use of transfusion practice guidelines. PMID- 15814290 TI - [Suitability of intraoperative autotransfusion]. AB - This work presents the procedure applied by our hospital to assess the quality and security of intra operative autotransfusion. The suitability of the three following variables has to be constantly assessed: performance of the machines to concentrate and wash collected blood, bacterial contamination of processed blood and rate of adverse events. We note that the procedure is applied with participation of medical and nursing staff. Since its setting-up, we note an amelioration of suitable variables. PMID- 15814291 TI - Serum protein media are important factors in the manual hexadimethrine bromide (polybrene) test, experience in China. AB - BACKGROUND AND OBJECTIVES: The use of the manual hexadimethrine bromide (polybrene) test in routine cross-matching after accurately detecting cell grouping and irregular antibodies is prevalent in China. This article reports the importance of serum protein mediums in the performance of the manual hexadimethrine bromide test. MATERIALS AND METHODS: Blood group O red blood cells and Blood group AB and Rh positive serum were collected at random from healthy blood donators, IgG anti-D serum separated from pregnant woman, then tested with each other by the manual hexadimethrine bromide methods in routine tests and some designed corresponding tests with IgG, IgM anti-D monoclonal diagnostic reagents and some serum protein components. RESULTS: Red blood cells that were adjusted to 3-5% suspension by normal saline then only added in 0.7 ml low ionic medium (LIM) and two drops of polybrene solution adhere to the surface of test tubes' bottom when centrifuged, so it was difficult to perform the next approach, but the adherence disappeared when red blood cells' concentrations exceeded 20-30%. Rh positive red blood cells coated by anti-D have the same phenomenon. This adherence can be prevented by serum medium diluted from 1:128 to 1:1024 times by normal saline and hemoglobin medium diluted from 1:32 to 1:128 times, but not by albumin or immunoglobulin medium. The denary logarithm values of the greatest inhibited dilutions of serum and hemoglobin elution between antibody sensitizing red blood cells and the same pre-sensitizing red blood cells tests were no significant difference (P value > 0.05). CONCLUSIONS: The whole serum or serum protein mediums are important factors that can influence successfully performance of the manual hexadimethrine bromide test. So appliance of the manual hexadimethrine bromide test to immunohematology laboratory, such as when performing titrations of serum or plasma, or when testing eluates for antibody activity, this adherence must be considered. PMID- 15814292 TI - [Theoretical risk management and legitimacy of the precautionary principle in medicine. Look back at HIV contamination through blood transfusion in France, twenty years ago]. AB - The precautionary principle first appeared in France during the health crisis following the contamination of patients with HIV via blood transfusion. This study analyses whether the risk associated with blood transfusion was taken into account early enough considering the context of scientific uncertainty between 1982 and 1985. The aim was to evaluate whether a precautionary principle was applied and whether it was relevant. First, we investigated the context of scientific uncertainty and controversies prevailing between 1982 and 1985. Then we analysed the attitude and decisions of the French authorities in this situation to determine whether a principle of precaution was applied. Finally, we explored the reasons at the origin of the delay in controlling the risk. Despite the scientific uncertainties associated with the potential risk of HIV contamination by transfusion in 1983, we found that a list of recommendations aiming to reduce this risk was published in June of that year. In the prevailing climate of uncertainty, these measures could be seen as precautionary. However, the recommended measures were not widely applied. Cultural, structural and economic factors hindered their implementation. Our analysis provides insight into the use of precautionary principle in the domain of blood transfusion and, more generally, medicine. It also sheds light on the expectations that health professionals should have of this principle. The aim of the precautionary principle is to manage rather than to reduce scientific uncertainty. The principle is not a futile search for zero risk. Rather, it is a principle for action allowing precautionary measures to be taken. However, we show that these measures must appear legitimate to be applied. This legitimacy requires an adapted decision-making process, involving all those concerned in the management of collective risks. PMID- 15814293 TI - [Foetomaternal erythrocyte incompatibilities: from immunohaematologic surveillance of pregnant women to haemolytic disease of the newborn]. AB - Despite the generalization of prevention measures against foetomaternal alloimmunization with anti-D immunoprophylaxis since 1970s, retrospectively 30 years later, its complications (new-born child's severe haemolytic disease, foetal death by anemia or nuclear icterus by bilirubin encephalopathy) have not disappeared. At the same time, alloimmunizations against antigens other than D increase with no possible prevention. As part of the set up in France of regional files analysing and making an inventory of serious foetomaternal incompatibilities requiring in utero or neonatal transfusion, we felt the need to synthesize current data, biological profiles (early screening of erythrocytic alloimmunization and its follow up during pregnancy, father's immunohaematologic status, evaluation of in utero immune haemolysis and impact of new non invasive techniques of diagnosis-RH1 foetal genotypage from ADN foetal of RH1--mothers' maternal plasma), clinical and paraclinical data (evaluation of foetal haemolysis by echography, recording of foetal movements and foetal cardiac rhythm), therapeutic indicators (in utero foetal transfusions or exsanguinotransfusions, neo and postnatal transfusions or exsanguinotransfusions, induced premature labour, newborn's intensive continue phototherapy and Rhesus immunoprophylaxis) in order to enable medical and paramedical professionals to carry out the specific supervision of pregnancies with foetomaternal incompatibility, the in utero, neo- and postnatal treatment of child and the efficient therapeutic prevention of anti-D alloimmunization, in a cooperative way. PMID- 15814294 TI - [Indications for transfusions of labile blood products]. AB - Indications for transfusions of red blood cells (RBC) are anemias, which can occur after trauma, in surgery, in obstetrics or oncohematology wards. The main criteria to administer RBC transfusion are hemoglobin level and clinical features. Transfusions are rare when the hemoglobin level is above 10 g/dL and are frequent when it is below 6 g/dL. However clinical setting, patient age, associated diseases, cardiovascular complications are taken into account. Immunocompatibility should always be tested and the transfusion consequences checked immediately and on the long term. Platelet transfusions are performed when the platelet count is low and patients suffered from hemorrhage. In oncohematology patients, platelet transfusion are administered with prophylaxis when the platelet count is lower than 10 g/L. Fresh frozen plasma has now a limited use, only in complex haemostatic disorders and in hemolytic uremic syndrome. PMID- 15814295 TI - [French training program for medical students in transfusion medicine. Transfusion Medicine Teachers' College]. AB - In France, transfusion medicine training program has been updated. A national committee of professors in transfusion medicine propose a series of 13 items which represent the minimum knowledge that general practitioners should possess. This overview of transfusion medicine is far below the level that specialists should reach and they will need an additional specialized training. Several French universities have set up their own training program which is quite similar to the work of the committee of professors. The following recommendations are not strict guidelines but is a common basis which will be improved in 2005 according to new evidence based transfusion medicine. PMID- 15814297 TI - Characterization of the human zinc finger protein 267 promoter: essential role of nuclear factor Y. AB - Liver fibrosis results from an excessive deposition of extracellular matrix proteins secreted by activated hepatic stellate cells (HSCs). The activation process is accompanied by an increased activity of various transcription factors, including zinc finger protein 267 (ZNF267). Recently, ZNF267 has been shown to modulate gene expression and to function as a transcriptional repressor. MMP-10 was identified as a target gene; its gene expression and promoter activity are inhibited by ZNF267, which might promote liver fibrogenesis through diminished matrix degradation. However, the transcriptional regulation of the ZNF267 gene is unknown. In the present study, we have cloned and characterized the human ZNF267 promoter containing a 1.5 kb fragment of the 5'-flanking region (-1414/+173). The ZNF267 gene has a TATA-less promoter with multiple transcription initiation sites. Analysis of serial 5'-deletions of luciferase reporter constructs revealed a minimal promoter between -72 and +173 bp. Mutational analysis of putative regulatory elements indicated that a CCAAT box within this region was essential for ZNF267 promoter activity. Electrophoretic mobility shift assays demonstrated that transcription factor nuclear factor Y (NF-Y) bound to the CCAAT box. In co transfection experiments, NF-YA increased the promoter activity of ZNF267. In conclusion, our results suggest that the binding site for NF-Y is critical for ZNF267 gene regulation and, herewith, the activation of this transcriptional factor may play an important role in the activation process of HSCs and in liver fibrosis. PMID- 15814296 TI - Circulating markers of oxidative stress and liver fibrosis in Sudanese subjects at risk of schistosomiasis and hepatitis. AB - Epidemiological studies in the developing world are frequently biased by the simultaneous presence of several infectious pathogens. In the present study, we examined the usefulness of circulating markers of oxidative stress and liver fibrosis to investigate the distinct forms of chronic liver inflammations associated with schistosomiasis and viral hepatitis, respectively. The study was performed in a Sudanese population exposed to Schistosoma. Circulating hyaluronic acid (HA) was used as a marker of liver fibrosis; the severity of schistosomiasis was determined by ultrasonic examination; viral hepatitis infection was ascertained by circulating anti-hepatitis antibodies. Serum markers were examined also in Sudanese subjects not exposed to Schistosoma infection and in French control subjects. We found a drastic decrease of lycopene levels in the subjects exposed to schistosomiasis in comparison with non-exposed Sudanese and French control subjects. Retinol, alpha-tocopherol and five carotenoids were unchanged. Lycopene depletion was unlikely to be due to variations of nutritional origin, since the lycopene/beta-carotene ratio was five-fold lower in the population at risk of schistosomiasis than in the other groups. We found that high HA serum levels were associated with severe periportal fibrosis but not with viral infection. Conversely, levels of the oxidized lipid malondialdehyde (MDA) were associated with viral infection but not with the severity of schistosomiasis, even though the two infections had additive effects. We concluded that serum markers are valuable tools for investigating the complex effects of co-existing factors of chronic liver inflammation. PMID- 15814298 TI - Characterization of the basal promoter element of the human type 5 17beta hydroxysteroid dehydrogenase gene. AB - Testosterone biosynthesis from androstenedione is carried out by androgenic 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity. Of the androgenic 17beta-HSD isoenzymes, only type 5 (17beta-HSD5) is expressed ubiquitously, including the human adrenal gland and ovary. To characterize this gene promoter, luciferase constructs of the human 5'-flanking region were transiently transfected into the H295R human adrenal carcinoma cell line. A series of fragment deletion constructs and electrophoretic mobility shift assays suggested that a sequence of CCTCCTCCT at -65 to -58 bp is the core sequence and demonstrated Sp1/Sp3 binding to this CCT repeat. Forskolin stimulated the promoter activity of the HSD17B5 gene through this Sp1/Sp3 binding site. Mutation of this site resulted in a significant reduction of HSD17B5 promoter basal and forskolin-induced activity. Mithramycin A, which inhibits the binding of Sp1 and Sp3 to DNA, also remarkably decreased HSD17B5 mRNA expression in the H295R cell line. These results indicate that members of the Sp family of transcription factors play an important role in regulating constitutive and stimulated expression of the HSD17B5 gene in H295R cells. PMID- 15814299 TI - New features of site-specific horseradish peroxidase (HRP) glycosylation uncovered by nano-LC-MS with repeated ion-isolation/fragmentation cycles. AB - Horseradish peroxidase (HRP) is widely used in biomedical research as a reporter enzyme in diagnostic assays. In addition, it is of considerable interest as a model glycoprotein with core-xylosylated and -(alpha1-3)-fucosylated N-glycans that form antigenic elements of plant allergens and parasitic helminths. Using a combination of techniques comprising (1) nano-liquid chromatography (LC)-mass spectrometry (MS)/MS with multiple selection/fragmentation cycles of HRP tryptic (glyco-)peptides, (2) nano-electrospray MS of intact HRP, and (3) carbohydrate linkage analysis, it was revealed that most of the HRP N-glycosylation sites can be occupied with an alternative Fuc(1-3)GlcNAc-disaccharide. Two main variants of HRP occur: The major population (approximately 60%) has eight glycosylation sites carrying core(1-3)fucosylated, xylosylated, trimannosyl N-glycans, with the ninth potential N-glycosylation site Asn316 not occupied. Another group of HRP carries seven of the above-mentioned N-glycans, with an eighth N-glycosylation site carrying the alternative Fuc(1-3)GlcNAc-unit (approximately 35%). In addition, minor subsets of HRP were found to contain a xylosylated, trimannosyl N-glycan lacking core-fucosylation as a ninth N-glycan attached to Asn316, which has hitherto been assumed to be unoccupied. The finding of these new features of glycosylation of an already exceptionally well-studied glycoprotein underscores the potential of the nano-LC-MS(n) based analytical approach followed. PMID- 15814300 TI - Expression of NOX-I, gp91phox, p47phox and P67phox in the aorta segments above and below coarctation. AB - Aorta coarctation results in hypertension (HTN) in the arterial tree proximal to stenosis and, as such, provides an ideal model to discern the effects of different levels of blood pressure on the vascular tissue in the same animal. Compelling evidence has emerged supporting the role of oxidative stress as a cause of HTN. However, whether or not HTN (independent of the circulating humoral factors) can cause oxidative stress is less certain. NAD(P)H oxidase isoforms are the main source of reactive oxygen species (ROS) in the vascular tissues. We therefore compared the expressions of NOX-I, gp91phox and the regulatory subunits of the enzyme in the aorta segments residing above and below coarctation in rats with abdominal aorta banding. Rats were studied 4 weeks after aorta banding above the renal arteries or sham operation. Subunits of NAD(P)H oxidase and its NOX-I isoform as well as endothelial NO synthase (eNOS) and nitrotyrosine (footprint of NO oxidation by superoxide) were measured in the aorta segments above and below coarctation. The gp91phox, p47phox, and p67phox subunits of NAD(P)H oxidase, NOX I isoform, eNOS and nitrotyrosine were markedly increased in the aorta segment above coarctation (hypertensive zone), but were virtually unchanged in the segment below coarctation. Since, excepting blood pressure, all other conditions were constant, the upregulation of NAD(P)H oxidase isoforms and the increased NO oxidation in the aorta segment above, but not below, coarctation prove that HTN, per se, independent of circulating mediators can cause oxidative/nitrosative stress in the arterial wall. These observations suggest that HTN control may represent a specific form of antioxidant therapy for hypertensive disorders. PMID- 15814301 TI - Soluble glucocorticoid-induced tumor necrosis factor receptor (sGITR) stimulates osteoclast differentiation in response to receptor activator of NF-kappaB ligand (RANKL) in osteoclast cells. AB - We found that treatment of osteoclast (OC) precursors with soluble glucocorticoid induced tumor necrosis factor receptor (sGITR) promoted osteoclastogenesis in the presence of macrophage colony-stimulating factor (M-CSF) and receptor for activation of nuclear factor-kappaB ligand (RANKL). Low levels of GITR and its ligand were expressed on the surface of OC precursor cells after incubation with RANKL. Stimulation of osteoclastogenesis by sGITR was blocked by neutralization with anti-GITR ligand antibody (Ab), whereas endogenous GITR did not affect osteoclastogenesis, indicating that enhancement of osteoclastogenesis by sGITR involves signaling via GITR ligand. The addition of sGITR decreased the level of interferon (IFN)-beta, and blockade of endogenous IFN-beta did not affect osteoclastogenesis stimulated by sGITR. We conclude that sGITR enhances osteoclastogenesis by acting on OC precursor cells to lower the level of IFN beta. PMID- 15814302 TI - Regulation of the human tissue-nonspecific alkaline phosphatase gene expression by all-trans-retinoic acid in SaOS-2 osteosarcoma cell line. AB - While tissue-nonspecific alkaline phosphatase (TNSALP) is a well-known indicator of bone formation and all-trans-retinoic acid a key regulator of that process, the relationship between TNSALP and retinoic acid has not yet been clearly described. The aim of the present study was therefore to clarify the mechanism by which retinoic acid modulates expression of TNSALP. After culturing SaOS-2 human osteoblastic osteosarcoma cells in the presence or absence of 10(-6) M all-trans retinoic acid, real-time RT-PCR confirmed that retinoic acid up-regulates expression of TNSALP mRNA. Notably, this time-dependent induction of TNSALP expression was accompanied by a corresponding increase in detected catalytic activity of the enzyme. When we then isolated the 5'-upstream region of the human TNSALP gene and carried out luciferase assays with a set of deletion mutants, we found that the promoter region contains a retinoic acid response element-like motif. Moreover, electrophoretic mobility shift assays showed that the nuclear extract bound to the motif. It thus appears that retinoic acid regulates the expression of human TNSALP via a retinoic acid response element in the genes promoter region. PMID- 15814303 TI - BMP-2 liberated from biomimetic implant coatings induces and sustains direct ossification in an ectopic rat model. AB - INTRODUCTION: Using a rat model, we evaluated the kinetics and histomorphometry of ectopic bone formation in association with biomimetic implant coatings containing BMP-2. MATERIALS AND METHODS: One experimental and three control groups were set up: titanium-alloy discs coated with a biomimetically co precipitated layer of calcium phosphate and BMP-2 [1.7 microg per disc (incorporated-BMP group)]; uncoated discs (control); discs biomimetically coated with a layer of calcium phosphate alone (control); and discs biomimetically coated with a layer of calcium phosphate bearing superficially adsorbed BMP-2 [0.98 microg per disc (control)]. Discs (n = 6 per group) were implanted subcutaneously in rats and retrieved at 7-day intervals over a period of 5 weeks for kinetic, histomorphometrical, morphological and histochemical analyses. RESULTS: In the incorporated-BMP-2 group, osteogenic activity was first observed 2 weeks after implantation and thereafter continued unabated until the end of the monitoring period. The net weekly rates of bone formation per disc were 5.8 mm3 at 2 weeks and 3.64 mm3 at 5 weeks. The total volumes of bone formed per disc at these junctures were 5.8 mm3 and 10.3 mm3, respectively. Bone tissue, which was formed by a direct ossification mechanism, was deposited at distances of up to 340 microm from the implant surfaces. The biomimetic coatings were degraded gradually, initially by foreign body giant cells alone and then also by osteoclasts. Forty percent of the coating material (and thus presumably of the incorporated BMP-2) remained at the end of the monitoring period. Hence, 60% of the incorporated BMP-2 had been released. At this 5-week juncture, no bone tissue was associated with any of the control implants. CONCLUSION: BMP-2 incorporated into biomimetic calcium phosphate coatings is capable not only of inducing bone formation at an ectopic site in vivo but also of doing so with a very high potency at a low pharmacological level, and of sustaining this activity for a considerable period of time. The sustainment of osteogenic activity is of great clinical importance for the osseointegration of dental and orthopedic implants. PMID- 15814304 TI - In vitro functional assay of alleles and haplotypes of two COL1A1-promoter SNPs. AB - Osteoporosis is a common disease with a strong genetic component. We previously described two polymorphic sites in the COL1A1 gene promoter, -1997 G/T and 1663indelT, which have been associated with bone mineral density (BMD), a surrogate trait for osteoporosis. Here, we explore the molecular mechanisms underlying this association by performing transient transfections in MG-63 cells of constructs bearing different COL1A1 promoter regions, containing different alleles or haplotypes of the polymorphic sites. These promoter regions drove the transcription of a luciferase reporter gene. The main differences in transcriptional activity relied on an inhibitory region localized to the -1284 to -254 interval. Regarding the polymorphisms, reproducible differences were observed between the alleles of each of them: the G allele at -1997 showed a higher transcriptional activity than the T allele, as did the 7T allele of -1663 as compared with 8T. Accordingly, the T-8T haplotype was the weakest transcriber. A functional interaction was found between the -1997 and -1663 polymorphisms, in that the difference in transcriptional activity between the 7T and 8T alleles was dependent on the allele at -1997. This different transcriptional activity of the two -1663indelT alleles correlated with different binding capacities of the corresponding oligonucleotides to osteoblast nuclear proteins. Supershift assays allowed us to identify one of these proteins as the architectural transcription factor Nmp4/CIZ, a protein known to be an inhibitor of BMP/Smad signalling. PMID- 15814305 TI - Composition of bone and apatitic biomaterials as revealed by intravital Raman microspectroscopy. AB - Microcharacterization of biominerals allows a better understanding of the pathophysiological events that occur in calcified tissues and synthetic biomaterials. Different methods have been extensively used to conduct such investigations. A new model for the intravital study of the composition and structure of membranous bone by Raman microspectroscopy is described. Titanium bone chambers equipped with a fused-silica optical window were implanted transcutaneously in the calvaria of New Zealand rabbits. The implanted optical windows were well tolerated, and spectral acquisitions were performed without any additional invasive procedure. Bone and implanted apatitic biomaterials were analyzed at different times after surgery. All Raman bands were unambiguously identified in the bone and biomaterial spectra. The main PO4 and CO3 Raman bands in bone spectra were consistent with those found in the carbonated apatite spectrum. The major collagen bands were always observed around 1200-1300 (amide III) and 1600-1700 (amide I) delta cm(-1) and, 1400-1470 and 2800-3100 delta cm( 1) (bending and stretching modes of CH groups, respectively). The phenylalanine (Phe) band was identified in all spectra at 1003 delta cm(-1) and overlapped that of the weak HPO4(2-) ion. The CH bands frequently overlapped the lipid bands. However a distinct protein and lipid bands were detected at 2950 and 2852 delta cm(-1), respectively. In bone areas close to blood vessels, the Raman signature of hemoglobin was detected with a characteristic band at 754 delta cm(-1). The changes observed in bone varied as a function of time and location. The composition and structure of all of the biomaterials studied--including those that were resorbable--seemed to remain stable over time and location. We report for the first time the complete intravital study of Raman spectra of bone and calcium phosphate biomaterials over a period of 8 months. This new approach does not require specimen preparation and allows simultaneous observation of mineral and organic bone constituents over time, which therefore should provide insightful information about their relationship. PMID- 15814306 TI - Effect of copper exposure during embryonic development on chemosensory function of juvenile fathead minnows (Pimephales promelas). AB - Fish rely on chemosensation to alert them of nearby predators. Recent evidence suggests that metals disrupt this chemical communication system. Our objective was to determine the chemical alarm response of juvenile fathead minnows after embryonic copper (Cu) exposure. Embryos were randomly assigned to one of two treatments: clean water or water containing 10 microg/L Cu. Once hatched, half of the Cu-exposed embryos were transferred to clean water (after hatch), while the other half remained in the Cu-contaminated water. Fish were tested using a triumvirate maze at the age of 84-96 d post-hatch. Fish reared in clean water significantly avoided the alarm cue. However, fish reared under continuous Cu exposure and those that were only exposed to Cu during embryonic development were unable to respond to the chemical alarm stimulus. Fish from all treatments did not respond to two control stimuli. Results from this study suggest that fish exposed to elevated Cu concentrations during embryonic development is sufficient to impair chemosensory function during later life stages. This could result in an inability to detect nearby predators by olfaction, which could lead to important ecological perturbations in populations inhabiting metal-contaminated systems. PMID- 15814307 TI - Selenium impacts on razorback sucker, Colorado River, Colorado I. Adults. AB - Adult razorback sucker (Xyrauchen texanus) were exposed to various selenium concentrations in ponds and isolated river channels of the Colorado River near Grand Junction, CO, to determine effects on their growth and residue accumulation over an 11-month period. Adults at Horsethief ponds were fed a commercial diet, whereas fish at Adobe Creek channel and North Pond foraged on natural food items. Selenium concentrations at Horsethief were 2.2 microg/L in water, 0.1-1.4 microg/g in sediment, and 2.3-3.1 microg/g in food organisms (1.1 microg/g in commercial fish food), at Adobe Creek were 3.8 microg/L in water, 0.5-2.1 microg/g in sediment, and 4-56 microg/g in food organisms, and at North Pond were 9.5 microg/L in water, 7-55 microg/g in sediment, and 20-81 microg/g in food organisms. The selenium concentrations in muscle plugs from adults at Adobe Creek (11.7 microg/g, SD = 0.4, n = 6) and North Pond (16.6 microg/g, SD = 1.0, n = 6) were greater than at Horsethief (4.5 microg/g, SD = 0.2, n = 6). During a depuration period adults from Adobe Creek and North Pond lost 1-2% of their selenium burden in 32 days and 14-19% in 66 days. Selenium accumulated in razorback sucker above toxic thresholds reported in other studies, yet those residues were less than those reported in muscle plugs of 40% of wild razorback sucker caught in the Green River, Utah. PMID- 15814308 TI - Selenium impacts on razorback sucker, Colorado River, Colorado II. Eggs. AB - Effects on hatching and development of fertilized eggs in adult razorback sucker (Xyrauchen texanus) exposed to selenium in flooded bottomland sites near Grand Junction, Colorado, were determined. After 9 months exposure, fish were collected and induced to spawn and eggs collected for inorganic element analyses. A 9-day egg study was conducted with five spawns from Horsethief ponds, six spawns from Adobe Creek channel, and four spawns from North Pond using a reference water and site waters. Selenium concentrations in eggs were 6.5 microg/g from Horsethief, 46 microg/g from Adobe Creek, 38 microg/g from North Pond, and 6.0 microg/g from brood stock. Eggs from young adults had a smaller diameter and higher moisture content than brood stock. There were no differences among the four sources in viability, survival, hatch, hatchability, or mortality of deformed embryos or larvae. Adobe Creek larvae had more deformed embryos in eggs held in site water than held in reference water. There were significant negative correlations between selenium concentrations in adult muscle plugs and percent hatch, egg diameter, and deformities in embryos. Results from this study suggest that selenium contamination in parts of the upper basin of the Colorado River should be a major concern to recovery efforts for endangered fish. PMID- 15814309 TI - Influence of a dispersant on the bioaccumulation of phenanthrene by topsmelt (Atherinops affinis). AB - Chemical dispersants enhance oil spill dispersion by forming water-accommodated micelles with oil droplets. However, how dispersants alter bioavailability and subsequent bioaccumulation of hydrocarbons is not well understood. Thus, the goal was to investigate the influence of a chemical dispersant on the disposition (uptake, biotransformation, and depuration) of a model hydrocarbon, [14C] phenanthrene ([14C]PHN), by larval topsmelt (Atherinops affinis). Exposure was via aqueous-only or combined dietary and aqueous routes from a water-accommodated fraction (WAF) of Prudhoe Bay Crude Oil (PBCO) or a WAF of Corexit 9527-dispersed PBCO (DO). Trophic transfer was measured by incorporating into exposure media both a rotifer (Brachionus plicatilis) as food for the fish and a phytoplankton (Isochrysis galbana) as food for the rotifers. Short-term (4 h) bioconcentration of PHN was significantly decreased in topsmelt when oil was treated with dispersant (P < 0.05), but differences diminished after 12 h. When trophic transfer was incorporated, PHN accumulation was initially delayed but after 12 h attained similar levels. Dispersant use also significantly decreased the proportion of biotransformed PHN (as 9-phenanthrylsulfate) produced by topsmelt (P < 0.05). However, overall PHN depuration was not affected by dispersant use. Thus, chemical dispersant use in oil spill response may reduce short-term uptake but not long-term accumulation of hydrocarbons such as PHN in pelagic fish. PMID- 15814310 TI - Ultraviolet damages sperm mitochondrial function and membrane integrity in the sea urchin Anthocidaris crassispina. AB - Effects of ultraviolet A (UVA) and ultraviolet B (UVB) on mitochondrial function and membrane integrity of sea urchin sperm were investigated using flow cytometry and fluorescent probes. Both UVA and UVB impaired sperm mitochondrial function in a dose-dependent manner. Covariance analysis further showed that the slopes of change in mitochondrial function in relation to UVA and UVB were significantly different, suggesting that the modes of action were different. UVA did not affect membrane integrity, while membrane integrity showed a linear reduction with increasing UVB doses. Sperm mitochondria function showed significant positive correlations with sperm motility and subsequent fertilization success. Overall, our results showed that both UVA and UVB could decrease sperm motility and fertilization success through impairment of mitochondrial function, whereas UVB alone could cause additional damage through impairing the functional integrity of sperm membrane. Mitochondrial function of sperm may also offer a reliable ecotoxicological biomarker for predicting fertilization success in urchins. PMID- 15814312 TI - The ability of dissolved organic matter (DOM) to influence benzo[a]pyrene bioavailability increases with DOM biodegradation. AB - The biodegradation of two substrates and the ability of dissolved organic matter (DOM) to influence benzo[a]pyrene bioavailability as DOM biodegradation progressed were monitored in reactors. Substrates were composed of algae extracts and an artificial substrate that mimics raw wastewater, which were considered to be autochthonous and anthropogenic allochthonous models for DOM, respectively. The soluble microbial products formed during biomass activity were also studied. The aromaticity of DOM was investigated with specific ultraviolet absorbance. Partitioning coefficients between DOM and benzo[a]pyrene, K(DOC)(biol), were biologically determined by means of 4-h bioaccumulation experiments on Daphnia magna. Parent and degraded substrates always significantly reduced the bioaccumulation of benzo[a]pyrene at environmental DOM concentrations. Soluble microbial products also significantly affected the benzo[a]pyrene bioaccumulation. K(DOC)(biol) ranged between 2 x 10(4) and 4 x 10(5) L/kg. As the artificial wastewater biodegraded, DOM aromaticity increased, as did K(DOC)(biol). During the biodegradation of algae extract DOM, K(DOC)(biol) increased, whereas their aromaticity slightly decreased. PMID- 15814311 TI - Evaluation of a constructed wetland treatment system specifically designed to decrease bioavailable copper in a wastestream. AB - A specifically designed constructed wetland decreased copper concentrations in a wastestream to 22 microg Cu/L and eliminated associated toxicity. Metal toxicity is a function of both concentration and form. This research measured copper partitioning to ligands within the wetland and observed changes in copper form with regard to bioavailability. Average monthly copper concentrations in the constructed wetland treatment system ranged from 10 to 47 microg/L in the upstream (i.e., inflow to the constructed wetland prior to the retention basin) and from non-detection to 11 microg/L in the downstream (i.e., outflow to the receiving stream). On average, 78% total-recoverable, 85% acid-soluble, and 83% soluble copper were removed from inflow to outflow of this constructed wetland; however, total recoverable and acid-soluble copper measurements were not useful indicators of bioavailable copper. Survival of Ceriodaphnia dubia increased from an average of 2% survival in the inflow to 96% in the outflow and reproduction increased from an average of 8 to 24 neonates/female, respectively. Soluble copper is a more accurate predictor of the concentration at which effects were observed. Average ratio of acid volatile sulfides to simultaneously extractable metals was 1.4: 0.06 micromol/g indicating sufficient sulfides to sequester available metals in the system. PMID- 15814313 TI - Effects of dehydroabietic acid and abietic acid on survival, reproduction, and growth of the crustacean Daphnia magna. AB - Resin acids, a class of wood extractives, are potential toxic constituents in many pulp and paper mill effluents. In the present investigation, the effects of two predominant resin acids, dehydroabietic acid (DHA) and abietic acid (ABA), on survival, reproduction, and growth of the freshwater crustacean Daphnia magna were assessed over its life cycle. Based on the experimentally determined acute toxicity data (48-h EC(50)'s) for DHA (7.48 mg/L) and ABA (7.98 mg/L), D. magna was treated chronically with each resin acid at nominal concentrations of 0, 0.25, 0.5, 1.0, 2.0, 4.0, and 8.0mg/L for 21 days. Both DHA and ABA at concentrations as high as 4.0mg/L did not affect physiological and reproductive parameters such as time to maturation, number of molting, number of broods, and number of offspring produced from surviving daphnids, while significant mortality was observed only at 8.0mg/L in both cases. However, a small but statistically significant decrease in Daphnia growth (body length) at the end of exposure was detected at concentrations as low as 0.5mg/L for DHA and 1.0mg/L for ABA, respectively. These results indicated that both DHA and ABA had the potential to exhibit weak growth inhibition without apparent negative effects on reproduction to D. magna at nonlethal concentration levels. This slight effect is not expected to be ecologically significant because the concentrations of DHA and ABA in biologically treated pulp and paper mill effluents are well below the effective levels observed in the present study. PMID- 15814314 TI - Impact assessment of various rearing systems on fish health using multibiomarker response and metal accumulation. AB - European sea bass were reared in three different systems: one flow-through (FTS), one recirculating (RAS), and one recirculating with a high-rate algae pond (RAS + HRAP). After 1 year of rearing, the final fish weight was 15% lower in the RAS compared to the FTS. The accumulation of a growth-inhibiting substance in the RAS is the main hypothesis explaining this difference. As in environmental risk assessment, fish bioaccumulation markers and biomarkers were used to demonstrate exposure to and effects of the rearing water in the three rearing systems. Thirty fish per system were sacrificed before their condition factor (CF) and liver somatic index (LSI) were calculated. Nine biomarkers, including ethoxyresorufin-O deethylase (EROD) and superoxide dismutase (SOD), were measured in liver and twelve metals including As, Cd, Cu, Pb, Cr, and Zn, for which there are regulations regarding human consumption, were measured in liver and muscle. In all systems, CF and LSI were not significantly different and no correlation was found with biomarker activity or metal concentration. EROD and SOD activities were significantly increased in RAS. Accumulation of seven and four metals in muscle and liver, respectively, was significantly higher in the RAS relative to FTS. The HRAP prevented metal accumulation except for chromium and arsenic. Eight metal concentrations were significantly higher in liver than in muscle. Concentrations of toxic metals were similar to reported values and below FAO/WHO recommended values for human consumption. PMID- 15814315 TI - Preliminary studies of biochemical changes (ethoxycoumarin O-deethylase activities and vitellogenin induction) in two species of shrimp (Farfantepenaeus duorarum and Litopenaeus setiferus) from the Gulf of Mexico. AB - Several investigations have demonstrated that the increase in chemicals in the environment may have caused effects on aquatic life and wildlife. The impact from oil production activities on the benthic community structure and shrimp (Farfantepenaeus duorarum and Litopenaeus setiferus) biochemical markers were studied in the Gulf of Mexico, which is heavily contaminated by polycyclic aromatic hydrocarbons and heavy metals. Shrimp were collected from a control area and from an area close to oil production platforms during October and November 2002. There was no spatial difference in exposure and response probably because shrimp migrate, as results did not show significant differences in cytochrome P4501A (as measured by ethoxycoumarin O-deethylase (ECOD) metabolism) between the two sites. In October, shrimp ECOD activities were higher and statistically different from those measured in the samples taken in November. As for ECOD activities, with the concentration of vitellogenin as another biomarker, there were no differences between the shrimp collected from the control area and the shrimp collected on the oil production platforms. In this case the concentrations were higher in shrimp collected in November vs. shrimp collected in October. However, there are significant correlations between contaminants and responses (biomarkers), indicating an effect of pollution. One of the most important considerations brought up by this kind of study is that, although the majority of groups studying the effects of endocrine disruption have focused almost exclusively on human health or vertebrate wildlife-related issues, it is necessary to increase research focused on understanding the function of hormones in invertebrate species exposed in the field to different kinds of pollutants. PMID- 15814316 TI - Mutagenicity assessment of effluents from textile/dye industries of Sanganer, Jaipur (India): a case study. AB - Sanganer town, district Jaipur (Rajasthan, India), is famous worldwide for its dyeing and printing industries. There are about 400 industries involved in textile printing processes, which discharge effluents into nearby ponds and drains, without any treatment. These effluents contain highly toxic dyes, bleaching agents, salts, acids, and alkalis. Heavy metals like cadmium, copper, zinc, chromium, and iron are also found in the dye effluents. Textile workers are exposed to such waters with no control over the length and frequency of exposure. Further, as the untreated effluents are discharged into the environment they can cause severe contamination of surface and underground water. Environmental pollution caused by such textile effluents results in adverse effects on flora, fauna, and the general health of not only the textile workers, but also the residents of Sanganer town. Therefore, to assess the possible genotoxic health risk and environmental genotoxicity due to the textile industry effluents, this study was carried out using the Ames Salmonella/microsome mutagenicity assay. The results clearly indicate that the effluents and the surface water of Amani Shah drainage have high mutagenic activity. Further, the drainage water and the dry bed of the drainage (during summer months) are not fit for agricultural or other recreational purposes. A low level of mutagenicity in the underground water of Sanganer again emphasizes the grave pollution problem existing in the area. Multiple post hoc comparison tests (LSD, Tukey's) were used for comparison of sample site, dose, and length of exposure. Quadratic Model was found to adequately fit the observed data. PMID- 15814317 TI - Acute toxicity bioassays of mercuric chloride and malathion on air-breathing fish Channa punctatus (Bloch). AB - Acute toxicity tests (96 h) were conducted in flow-through systems to determine the lethal toxicity of a heavy metal compound, mercuric chloride, and an organophosphorus pesticide, malathion, to air-breathing teleost fish, Channa punctatus (Bloch) and to study their behavior. The 96-h LC50 values were determined, as well as safe levels. The results indicate that mercuric chloride is more toxic than malathion to the fish species under study. Dose- and dose-time dependent increases in mortality rate were also observed in response to both test chemicals. PMID- 15814318 TI - The immunomodulatory effects of the dimer of lysozyme (KLP-602) in carp (Cyprinus carpio L)--in vivo study. AB - The dimer of lysozyme was administered to carp (Cyprinus carpio L.) in an attempt to reduce the suppression of response in fish exposed to the cypermethrin. Experimental studies were carried out on cells isolated from blood and pronephros of carp. In the in vivo studies fish were intoxicated with cypermethrin (synthetic pesticide) at a dose of LD5. After 24 h fish were injected intraperitoneally with a modulator-the dimer of lysozyme (KLP-602) at a dose of 0.02 mg/kg of body weight. In the study we determined the metabolic activity of phagocyte cells and the proliferative ability of lymphocytes stimulated by ConA and LPS. The results indicate that this modulator is useful for stimulation of cellular and humoral immunity after experimentally induced suppression by selected pesticide. PMID- 15814319 TI - Evaluation of cytogenetic effects of lambda-cyhalothrin on Wistar rat bone marrow by gavage administration. AB - In this study, a synthetic pyrethroid insecticide, lambda-cyhalothrin (LCT), was administered to adult female albino rats (Wistar rats) by gavage dose of 6.12, 3.06, 0.8 mg/kg b.w. repeated for 13 days at 48 h intervals. The cytotoxic and genotoxic effects of LCT were investigated in bone marrow cells, using the structural chromosomal aberration (SCA) and micronucleus (MN) test systems. Mitomycin C (MMC) was also used as positive control (2mg/kg b.w.). All the doses of LCT increased the number of SCAs and the frequency of micronucleated erythrocytes, with respect to the control group. Only the highest dose of LCT significantly increased the MN frequency compared with control (P < 0.01). It was also observed that LCT caused a significant decrease in the number of polychromatic erythrocytes compared with controls (p < 0.001). These observations indicate the in vivo suspectibility of mammals to the genetic toxicity and cytotoxicity potential of LCT. PMID- 15814320 TI - Liver phosphatases in mice treated with lead during murine ancylostomiasis. AB - Both infective larvae and pollutants disturb the homeostasis of female Swiss albino mice, causing ancylostomiasis and severe immunophysiological changes. In our experimental design, mice in group A were fed 0.01 mg of lead nitrate before infection with 500 larvae of Ancylostoma caninum, mice in group B were infected with larvae without previous lead treatment, and group C received lead nitrate treatment alone. Control animals in group D were neither treated with lead nitrate nor infected with larvae. While all three experimental groups showed disturbances in liver alkaline phosphatase (ALP) and acid phosphatase (ACP), group C receiving lead nitrate treatment alone and group A receiving lead nitrate with infection showed increased levels of ACP and decreased levels of ALP. PMID- 15814322 TI - [Depression: an underrated consequence of stroke in elderly]. AB - Stroke is a major cause of morbidity, disability and hospitalization in the elderly. Depression frequently occurs after stroke and influences functional recovery, a crucial factor for the prognosis. The physiopathology of post-stroke depression is not entirely elucidated and might involve several mechanisms: direct consequences of brain lesions, especially in certain localizations, neuroendocrine mechanism or psychological reaction to a life event responsible for stress and handicap. Antidepressant drugs improve depressive symptoms and functional recovery. Therefore, search for depression should be systematic early at the stroke reeducation phase to instaure appropriate treatment. PMID- 15814323 TI - [Transient ischemic attacks in the elderly: new definition and diagnostic difficulties]. AB - Transient ischemic attacks (TIA) are very frequent in the elderly. Their frequency increases beyond 65 years. However, no epidemiologic study was specifically dedicated to elderly patients. The first definition of TIA was a sudden focal neurologic deficit that lasted for less than 24 hours, presumed to be of vascular origin and located in a specific artery territory of the brain or eye. The Working Study Group has proposed a new definition: TIA is a brief episode of neurologic dysfunction caused by focal brain or retinal ischemia with clinical symptoms typically lasting less than one hour, most often some minutes, and without evidence of acute infarction. Weighted diffusion MRI may show very early an aspect of cytotoxic oedema. The one-hour criterion associated with a stable neurological deficit is requested for initiating IV thrombolysis, if the angio-MRI shows an occlusion of the supra-aortic trunks or intracranial arteries, even in aged patients. Each TIA constitutes a major risk for a completed infarct resulting in disability or death. Hypertension is the main risk factor for TIAs, followed by atrial fibrillation, diabetes, coronaropathy and sedentarity. These factors multiply by 4 the stroke risk. In the elderly, TIAs are pecularly associated with lacunar infarcts in the territory of deep perforating arteries. TIAs represent a neurologic emergency that allows no delay in clinical and laboratory investigations, such as ultrasonic echographies and weighted diffusion MRI. Diagnostic errors are often due to frequent polypathology and cognitive changes in great age. The most misleading symptoms are vertigo, imbalance, falls, disorders of consciousness. Unawareness of the deficit is also a frequent cause of failure of TIA diagnosis. Conversely, the most frequent cause of diagnostic error by excess is epileptic seizures which are often under-evaluated. PMID- 15814324 TI - [Useful assessment for identifying unsafe driving]. AB - Ability to drive safely is the resultant of interactions between the individual (the driver), the vector (the car) and the environment (the state of the road). For some aged drivers, an important decline of visual, musculosquelettic and cognitive performances, may affect the ability to drive and increase the rate of crashes per vehicle-kilometer-driven, and the morbidity and mortality related to crash. Therefore, each holder of a driving licence should be medically suited to control his driving ability. In case of transitory or lasting driving incapacity, drivers must, of their own initiative, stop driving according to the Highway code and the contractual obligations appearing in their vehicle insurance contract. A medical examination for aptitude to driving requires a standardized, reliable, reproducible procedure based on consensual assessment tools to avoid arbitrary decisions for driving cessation. We propose a multidisciplinary approach to detect important decline of visuospatial and motor skills, paroxystic drops of attention and vigilance, and decreased cognitive capacity to anticipate and adapt driving at every moment. This assessment is based on a semi-directed interview and simple diagnostic tests. According to the present French law, only twelve medical conditions or functional deficits are inconsistent with the retain of the driving licence for a light vehicle. PMID- 15814325 TI - [Retiring: a test of resilient capacities]. AB - This article deals with the question of retiring as a test of the conjunctural resilience capacities. The continuity theory and the rupture theory are developed because their content is related to protective factors and risk factors met by retirees: the first theory advocates the use of familiar strategies in familiar areas to maintain internal and external structures. This continuity would protect mental health. The second one emphasises the gap with the working world and its affiliated status. The loss of the professional status would endanger, in a way more or less important, the subjects' mental health. Retiring, as a life event, can be experienced as a trauma or can even be compared by the subjects themselves to other significant periods of their life which have been more or less well overcome. In order to attempt to figure out the conjunctural resilience capacities requested on the first year of retirement, and at the same time, highlighting the available protective factors, two clinical vignettes of two potentially young resilient retirees are presented. PMID- 15814326 TI - ["Normal pressure" hydrocephalus]. AB - Normal pressure hydrocephalus (NPH) or, more precisely, chronic adult hydrocephalus, is a complex condition. Even if the basic mechanism is found in an impediment to CSF absorption, the underlying pathology is heterogeneous. In secondary NPH, the disruption of normal CSF pathways, following meningitis or sub arachnoid haemorrhage, is responsible for ventricular dilatation. However, in about half of the cases, the etiology remains obscure. NPH is more frequently found in elderly people, probably in relation with the increased incidence of cerebrovascular disease. The diagnosis of NPH is based upon a triad of clinical symptoms. The main symptom is gait disturbances, followed by urinary incontinence and various degree of cognitive changes. The latter two symptoms are not prerequisites for the diagnosis. Radiological ventricular dilatation without cortical sulcal enlargement is a key factor, as well as substantial clinical improvement after CSF withdrawal (CSF tap test). Other CSF dynamic studies and various imaging investigations have been proposed to improve diagnostic accuracy, but no simple test can predict the results of CSF drainage. The current treatment is ventriculo-peritonial shunting, ideally using an adjustable valve. Results are directly dependent upon the accuracy of the preoperative diagnosis. Post-surgical complications may be observed in about 10% of cases. PMID- 15814327 TI - [Role of personality in depression of the elderly: difference between early and late life depression]. AB - Personality disorders have been implicated in the occurrence of depression in the elderly. The main purpose of this study was to assess the role of personality disorders in depression of the elderly and to distinguish between early and late onset depression. The study included 48 subjects over 65 years of age from a department of psychiatry, who suffered from a major depressive episode according to the criteria of the DSM-III-R, without bipolar characteristics. The patients were examined at two different times. At the first interview, depression was assessed by the mini-GDS and the CES-D scales, and a cognitive disorder was ruled out by the Mini-Mental State Examination. The patients were then classified in two groups according to the time of the first occurrence of depression, before (early onset depression) or after (late onset depression) 65 years of age. A second evaluation was performed after the cure of the depression. The patients' personality was then assessed using the International Personality Disorder Examination, in its VKP French-translated version, which evaluates personality disorders as defined by the criteria of the DSM-III-R and the ICD-10. The frequency of personality disorders was higher in patients with early-onset depression rather than in those with late onset depression. The most frequent personality disorder was avoiding personality (Cluster C) according to categorical as well as dimensional assessment. "Dependant personality" (Cluster C) was also quite often associated with early-onset depression. However this results should be confirmed by a larger study. PMID- 15814328 TI - Antigen recognition and presentation by dendritic cells. AB - In this article we review the following important points in the antigen presenting system: (1) the regulation of the expression of major histocompatibility complex (MHC) molecules, (2) the mechanism of cross presentation, and (3) the interaction of antigen-presenting cells (APC) and T cells. 1. The expression of MHC class I or class II molecules is regulated by the interaction of the MHC enhanceosome and the class II transactivator (CIITA). CIITA also regulates the gene expression of plexna-1, which encodes a semaphorin receptor, plexin-A1, that might be involved in the interaction with T-cells through an unknown ligand for plexin-A1. 2. Two pathways, a proteasome/TAP independent pathway and a proteasome/TAP-dependent pathway, have now been identified in the cross-presentation. In the proteasome/TAP-dependent pathway, the translocon/Sec61 protein channel is an important element for the transport of antigenic peptides in phagosomes to the cytoplasm. 3. The integration of adhesion/costimulatory molecules and peptide-MHC complexes at the surface of APC creates the "immunological synapse" region, which potentiates the efficiency of APC-T-cell interactions. The peptide-MHC complexes preferentially reside in the "raft" structure or associate with tetraspanin family molecules. PMID- 15814329 TI - Functional diversity and plasticity of human dendritic cell subsets. AB - The induction of different types of innate and adaptive immune responses, depending on the nature of the antigens and the environmental context, is crucial to cope with a variety of pathogens and concurrently to avoid pathologic reaction to self antigens. Recent studies have elucidated that the diversity of immune responses is critically controlled by dendritic cells (DCs). Two DC subsets, myeloid DCs and plasmacytoid DCs, have been identified in humans. The DC subsets recognize different microbial pathogens by expressing distinct repertoires of Toll-like receptors and induce different types of innate and adaptive immune responses, depending on the environmental factors. In particular, plasmacytoid DC precursors produce vast amounts of type I interferons in response to viruses and thus play an important role in antiviral immunity. Elucidating the cellular and molecular mechanisms that modulate the functions of the 2 DC subsets will lead to an understanding of the pathogenesis of various immune-related diseases and to the development of novel immunologic therapies. PMID- 15814330 TI - Dendritic cells: sentinels of immunity and tolerance. AB - The induction of effective antigen-specific T-cell immunity to pathogens without the initiation of autoimmunity has evolved as a sophisticated and highly balanced immunoregulatory mechanism. This mechanism assures the generation of antigen specific effector cells as well as the induction and maintenance of antigen specific tolerance to self-structures of the body. As professional antigen presenting cells of the immune system, dendritic cells (DC) are ideally positioned throughout the entire body and equipped with a unique capability to transport antigens from the periphery to lymphoid tissues. There is growing evidence that DC, besides their well-known immunostimulatory properties, also induce and regulate T-cell tolerance in the periphery. This regulatory function of DC is strictly dependent on their different stages of maturation and activation. Additionally, immunosuppressive agents and cytokines further influence the functions of maturing DC. The regulatory properties of DC include induction of T-cell anergy, apoptosis, and the generation of T-cells with regulatory capacities. This brief review summarizes the current knowledge about the immunoregulatory role of DC as guardians for the induction of T-cell immunity and tolerance. PMID- 15814331 TI - Migration of dendritic cells. AB - The migration of dendritic cells (DCs) to lymph nodes (LNs) is pivotal to the establishment of the immune response. DCs have been proved to pass through the afferent lymphatic pathway to enter LNs from the peripheral tissues after they have scanned for self or nonself antigens. In response to danger signals, both myeloid and plasmacytoid DC precursors (mDC and pDC precursors) are rapidly mobilized into the circulation. mDC precursors are recruited to inflamed tissues in response to inflammatory chemokines and then remobilized to regional LNs in response to CCL21. In contrast, pDC precursors directly transmigrate to regional LNs via high endothelial venules in a CXCL9- and E-selectin-dependent manner. Such migration is largely dependent on systemic inflammatory reactions. After accumulating in the LNs through distinct trafficking pathways, DCs interact with lymphocytes temporally and spatially to establish effective immune responses. The inflammation-dependent, chemokine-driven property of DC precursor trafficking is a very sophisticated host defense system. PMID- 15814332 TI - In vivo platelet production from mature megakaryocytes: does platelet release occur via proplatelets? AB - Although platelets are universally accepted to be born from megakaryocytes (MKs), the mechanism by which platelets are formed and released from MKs in vivo remains controversial. One theory, known as the proplatelet theory, postulates that platelets are released from proplatelet processes protruding from MKs into sinusoids located in the bone marrow hematopoietic compartment. Proplatelet formation (PPF) has been observed in in vitro experiments involving detailed analyses of related molecular events. PPF has also been used as a marker of MK maturation. However, PPF is suggested to be a nonphysiological phenomenon. On the other hand, transmission electron microscopy (TEM) analyses have revealed platelet formation via explosive fragmentation of MK cytoplasm in bone marrow and lung capillaries prepared by immersion fixation. Moreover, TEM and scanning electron microscopy studies of liquid-cultured MKs kept in suspension show that platelet formation occurs without PPF. Rather, an explosive and global fragmentation of the MK cytoplasm composed of platelet territories has been reported as the mechanism of platelet formation. In addition, in vivo and ex vivo observations of platelet release from MKs with phase-contrast microscopy strongly support the explosive-fragmentation theory. With all observations taken into account, PPF may not be a prerequisite for platelet release from MKs under real life conditions. In this review, a new "protoplatelet" concept is proposed to support the explosive-fragmentation theory. Additionally, the role of the lungs in platelet production is reviewed and discussed. PMID- 15814333 TI - Nitric oxide derived from human umbilical vein endothelial cells inhibits transendothelial migration of neutrophils. AB - We evaluated the roles of nitric oxide (NO) derived from endothelial cells in neutrophil transendothelial migration (TEM). Pretreatment of human umbilical vein endothelial cells (HUVECs) with NG-nitro-L-arginine methyl ester hydrochloride (L NAME) or NG-monomethyl L-arginine (L-NMMA), which are inhibitors of NO synthases, enhanced neutrophil TEM. Similar augmentation of TEM was observed in the presence of an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3 oxide (carboxy PTIO). Neutrophil TEM across L-NAME- or L-NMMA-treated HUVECs was inhibited by continuous NO supply by NO donors. These findings support the suggestion that continuous production of NO by endothelial cells suppresses neutrophil TEM. Flow cytometric analyses revealed that NO accumulates in neutrophils co-cultured with NO-producing HUVECs. A decreased amount of NO was detected in neutrophils co-cultured with L-NAME-treated HUVECs compared with neutrophils co-cultured with untreated HUVECs. Soluble guanylyl cyclase (sGC) is known as one of the most important targets of NO in neutrophils. 3-(53 Hydroxymethyl-23furyl)-1-benzyl indazole (YC-1), an activator of sGC, inhibited L NAME-induced neutrophil TEM. It was interesting that inhibition of neutrophil sGC with 1-H[1,2,4-]oxadiazolo[4,3-a]quinoxalin-1-1 (ODQ) was sufficient to enhance TEM. These results suggest that NO derived from HUVECs acts on neutrophils to inhibit TEM, at least in part by activating sGC. Our findings imply the role of NO constitutively generated by HUVECs in protection against excessive neutrophil extravasation and unnecessary tissue damage under physiological conditions. PMID- 15814334 TI - Fluorescence in situ hybridization monitoring of BCR-ABL-positive neutrophils in chronic-phase chronic myeloid leukemia patients during the primary stage of imatinib mesylate therapy. AB - We describe a method for monitoring chronic myeloid leukemia (CML) patients treated with imatinib that uses fluorescence in situ hybridization (FISH) to detect BCR-ABL in peripheral blood (PB) granulocytes. First, we compared this method, termed Neutrophil-FISH, with interphase FISH (i-FISH) analysis of bone marrow (BM), i-FISH analysis of PB mononuclear cells, and conventional cytogenetic analysis (CCA) of BM in 30 consecutive CML patients. We found the percentage of BCR-ABL-positive neutrophils as determined by Neutrophil-FISH to correlate best with the percentage of Philadelphia chromosome-positive metaphases in the BM determined by CCA (y = 0.8818x + 5.7249; r(2) = 0.968). We then performed a serial Neutrophil-FISH study of 10 chronic-phase CML patients treated with imatinib and found that the technique could clearly separate imatinib responders from nonresponders within 12 weeks of drug administration. There was a significant difference in the percentages of BCR-ABL-positive neutrophils between responder (mean 3 SD, 18.2% 3 11.8%) and nonresponder (82.4% 3 5.1%) groups at 12 weeks (P < .0001, Student t test).Together with real-time quantitative polymerase chain reaction analysis, Neutrophil-FISH represents another useful method for monitoring CML patients during the primary myelosuppressive stage of imatinib therapy because it is a quick, simple, and reliable method for assessing cytogenetic response. PMID- 15814335 TI - Successful treatment with imatinib mesylate in a case of minor BCR-ABL-positive acute myelogenous leukemia. AB - Philadelphia (Ph) chromosome-positive acute myelogenous leukemia (AML) is a rare disease that is resistant to conventional antitumor chemotherapy and has a poor prognosis. We describe a case of Ph chromosome-positive AML in which imatinib mesylate was used and a favorable outcome was obtained.A 64-year-old man was found to have Ph chromosome-positive, minor BCR-ABL-positive AML. Remission could not be induced by remission induction therapy with antitumor agents. Because the patient had a serious concomitant infectious disease, administration of 600 mg/day of imatinib mesylate, a specific inhibitor of BCR-ABL tyrosine kinase, was started after written informed consent was obtained. Complete cytogenetic response (CCR) was achieved without serious adverse events and persisted for more than 1 year. Our results suggested that imatinib mesylate was very useful for treating Ph chromosome-positive AML. PMID- 15814336 TI - Long-term results of a multicenter randomized, comparative trial of modified CHOP versus THP-COP versus THP-COPE regimens in elderly patients with non-Hodgkin's lymphoma. AB - In treating elderly non-Hodgkin's lymphoma (NHL) patients, it is particularly important to use drugs that have a low incidence of adverse events and high efficacy. In this multicenter study, THP (pirarubicin)-COP (cyclophosphamide, vincristine, and prednisolone) was compared to two thirds dosage of full CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone) regimen with regard to both adverse events and efficacy. For a third group, etoposide (E) was added to the THP-COP regimen (THP-COPE) in order to achieve high dose-intensity. Subjects were 486 previously untreated patients, aged 65 or older (range, 65-92 years; median, 74 years), with NHL. Subjects were randomly assigned to receive THP-COP, two thirds CHOP, or THP-COPE. Four hundred and forty-three patients were assessed for response and followed for 8 years after the last subject registered. The complete remission rates for the THP-COP, CHOP, and THP-COPE groups were 42.5%, 41.4%, and 48.0%, respectively. There was no difference in overall survival or progression-free survival among these 3 groups. In aggressive lymphoma, there was also no difference in complete response (CR) rate (45.3% in THP-COP, 44.9% in CHOP, 48.0% in THP-COPE), overall survival, and progression free survival among these groups. The 5- and 8-year survival rates for all patients were 29.4% and 18.7%, respectively. The 5- and 8-year survival rates for patients with aggressive lymphoma were 27.4% and 17.4%, respectively. Although long-term survival for patients with aggressive lymphoma on our regimens was not worse compared to previous reports, the CR rate was lower. Because severe adverse events were not observed, higher dose chemotherapy may be directed to achieve better CR rates. In patients with T-cell-type lymphoma, the CR rate was greater after treatment with THP-COP (51.4%) or THP-COPE (57.7%) compared to treatment with CHOP (19.4%). Pirarubicin may be more useful for T-cell lymphoma than doxorubicin. Because adverse cardiac events were reported only in CHOP, adverse cardiac events might be low in the THP group. PMID- 15814337 TI - Plasmacytoma of the urinary bladder in a renal transplant recipient. AB - A 28-year-old woman underwent renal transplantation in 1993. Eight years later, she experienced macroscopic hematuria, and Epstein-Barr virus-negative solitary extramedullary plasmacytoma (EMP) of the urinary bladder was diagnosed. After the reduction of immunosuppressive therapy, she received combined chemotherapy, resulting in complete tumor disappearance. However, 10 months later, she relapsed with aggressive multiple EMP and died of disease progression in 2003. This report is the first of a case of solitary EMP of the urinary bladder appearing as posttransplantation plasma cell dyscrasias after renal transplantation. PMID- 15814338 TI - Influential factors for the collection of peripheral blood stem cells and engraftment in acute myeloid leukemia patients in first complete remission. AB - Although several studies have investigated factors influencing peripheral blood stem cell (PBSC) mobilization in patients with nonmyeloid malignancies in an effort to increase the efficiency of autologous PBSC transplantation (APBSCT), there are very few reports on the efficiency of PBSC mobilization in patients with leukemia. We analyzed the effects of influential variables on successful mobilization and the correlation between infused cell doses and engraftment in acute myeloid leukemia (AML) patients in first complete remission (CR1) who received APBSCT. Between May 1998 and May 2003, 34 patients with AML underwent APBSC collections at our institution. All patients were in CR1 at the time of transplantation. Except for 1 patient, all patients successfully achieved the target CD34(+) cell yield of > or = 2 x 10(6)/kg. Among progenitor cells, the CD34(+) cell dose and the colony-forming unit-granulocyte-macrophage count showed significant correlations with neutrophil and platelet engraftments. The time to neutrophil engraftment was inversely correlated to the number of infused CD34(+) cells (r = -0.67; P < .001), whereas the time to neutrophil engraftment was not significantly correlated with the number of monocytes (r = 0.20; P = .701) or the number of nucleated cells (r = 0.35; P = .062). The time to platelet engraftment was significantly correlated with the dose of infused CD34(+) cells (r = -0.47; P = .012). The univariate analysis showed that more CD34(+) cells per kilogram and more CD34(+) cells per kilogram per day were collected from patients who had a shorter interval (less than 2 months) between diagnosis and PBSC harvest (P = .0111). In conclusion, this study showed that the CD34(+) cell dose was most strongly correlated with a successful engraftment in AML CR1 patients who underwent APBSCT. The proper timing of PBSC collections should be explored to optimize the outcome of APBSCT in AML CR1 patients. PMID- 15814339 TI - Tacrolimus-related encephalopathy following allogeneic stem cell transplantation in children. AB - Tacrolimus is a potent immunosuppressive drug widely used to prevent and treat graft-versus-host disease (GVHD) in stem cell transplantation (SCT). Among 49 patients receiving tacrolimus who underwent SCT from January 2000 to July 2003, 10 patients (20%) developed encephalopathy. The commonly observed symptoms were convulsions and drowsiness, and most patients complained of signal symptoms such as headache, nausea, and cortical blindness before onset. The most common abnormality on neuroimages was high-intensity lesions in white matter on magnetic resonance imaging T2-weighted or fluid-attenuated inversion recovery images. At onset, all patients were receiving treatment for acute GVHD (grade II/III) or extensive chronic GVHD and demonstrated an abrupt increase in blood pressure from baseline levels. The serum tacrolimus concentration was generally within acceptable levels at onset. Symptoms gradually improved in all patients when the blood pressure was lowered with antihypertensive medication, regardless of continued tacrolimus administration following a short-term suspension. The pathogenesis of tacrolimus-related encephalopathy is multifactorial, although refractory GVHD and a sudden increase in blood pressure seem to be major predisposing factors. Because the withdrawal of tacrolimus or switching to less potent anti-GVHD agents usually worsens the GVHD, the administration of tacrolimus should be managed by closely monitoring serum levels and controlling blood pressure. PMID- 15814340 TI - Trisomy 22 as the sole karyotypic abnormality in myelodysplastic syndrome. PMID- 15814341 TI - The 2010 challenge: data availability, information needs and extraterrestrial insights. AB - At the 2002 Johannesburg World Summit on Sustainable Development, 190 countries endorsed a commitment to achieve, by 2010, a significant reduction of the current rate of biodiversity loss at the global, regional and national levels. A wide range of approaches is available to the monitoring of progress towards this objective. The strengths and weaknesses of many of these approaches are considered, with special attention being given to the proposed and existing indicators described in the other papers in this issue. Recommendations are made about the development of indicators. Most existing and proposed indicators use data collected for other purposes, which may be unrepresentative. In the short term, much remains to be done in expanding the databases and improving the statistical techniques that underpin these indicators to minimize potential biases. In the longer term, indicators based on unrepresentative data should be replaced with equivalents based on carefully designed sampling programmes. Many proposed and existing indicators do not connect clearly with human welfare and they are unlikely to engage the interest of governments, businesses and the public until they do so. The extent to which the indicators already proposed by parties to the Convention on Biological Diversity are sufficient is explored by reference to the advice an imaginary scientific consultant from another planet might give. This exercise reveals that the range of taxa and biomes covered by existing indicators is incomplete compared with the knowledge we need to protect our interests. More fundamentally, our understanding of the mechanisms linking together the status of biodiversity, Earth system processes, human decisions and actions, and ecosystem services impacting human welfare is still too crude to allow us to infer reliably that actions taken to conserve biodiversity and protect ecosystem services are well chosen and effectively implemented. The involvement of social and Earth system scientists, as well as biologists, in collaborative research programmes to build and parameterize models of the Earth system to elucidate these mechanisms is a high priority. PMID- 15814342 TI - Monitoring global rates of biodiversity change: challenges that arise in meeting the Convention on Biological Diversity (CBD) 2010 goals. AB - By agreeing to strive for 'a significant reduction in the current rate of loss of biological diversity' by the year 2010, political leaders at the 2002 World Summit on Sustainable Development (held in Johannesburg, South Africa) presented conservation scientists with a great opportunity, but also one of their most significant challenges. This is an extremely exciting and laudable development, but this reporting process could be made yet more powerful if it incorporates, from the outset, independent scientific assessment of the measures, how they are analysed, and practical ways of plugging key gaps. This input is crucial if the measures are to be widely owned, credible and robust to the vigorous external scrutiny to which they will doubtless be exposed. Assessing how rates of biodiversity loss have changed from current levels by 2010 will require that a given attribute has been measured at least three times; however, most habitats, species, populations and ecosystem services have not been assessed even once. Furthermore, the best data on which to base estimates of biodiversity loss are biased towards the charismatic vertebrate species; unfortunately, these supply minimal services to the human economy. We have to find ways to redress this taxonomic imbalance and expand our analyses to consider the vast diversity of invertebrate, fungal and microbial species that play a role in determining human health and economic welfare. In the first part of this paper I will use examples from local and regional monitoring of biological diversity to examine the desired properties of 'ideal indicators'. I will then change focus and examine an initial framework that asks how we might monitor changes in the economic goods and services provided by natural ecosystems. I will use this exercise to examine how the set of possible indicators given by the Convention on Biological Diversity might be modified in ways that provide a more critical assay of the economic value of biological diversity. Here I will emphasize that we need not only to monitor these benefits, but also to significantly increase public awareness of human dependence upon the role that non-voting species play in driving the world's financial economy. PMID- 15814343 TI - Monitoring change in biodiversity through composite indices. AB - The need to monitor trends in biodiversity raises many technical issues. What are the features of a good biodiversity index? How should trends in abundance of individual species be estimated? How should composite indices, possibly spanning very diverse taxa, be formed? At what spatial scale should composite indices be applied? How might change-points--points at which the underlying trend changes- be identified? We address some of the technical issues underlying composite indices, including survey design, weighting of the constituent indices, identification of change-points and estimation of spatially varying time trends. We suggest some criteria that biodiversity measures for use in monitoring surveys should satisfy, and we discuss the problems of implementing rigorous methods. We illustrate the properties of different composite indices using UK farmland bird data. We conclude that no single index can capture all aspects of biodiversity change, but that a modified Shannon index and the geometric mean of relative abundance have useful properties. PMID- 15814344 TI - Using Red List Indices to measure progress towards the 2010 target and beyond. AB - The World Conservation Union (IUCN) Red List is widely recognized as the most authoritative and objective system for classifying species by their risk of extinction. Red List Indices (RLIs) illustrate the relative rate at which a particular set of species change in overall threat status (i.e. projected relative extinction-risk), based on population and range size and trends as quantified by Red List categories. RLIs can be calculated for any representative set of species that has been fully assessed at least twice. They are based on the number of species in each Red List category, and the number changing categories between assessments as a result of genuine improvement or deterioration in status. RLIs show a fairly coarse level of resolution, but for fully assessed taxonomic groups they are highly representative, being based on information from a high proportion of species worldwide. The RLI for the world's birds shows that that their overall threat status has deteriorated steadily during the years 1988 2004 in all biogeographic realms and ecosystems. A preliminary RLI for amphibians for 1980-2004 shows similar rates of decline. RLIs are in development for other groups. In addition, a sampled index is being developed, based on a stratified sample of species from all major taxonomic groups, realms and ecosystems. This will provide extinction-risk trends that are more representative of all biodiversity. PMID- 15814345 TI - Developing indicators for European birds. AB - The global pledge to deliver 'a significant reduction in the current rate of biodiversity loss by 2010' is echoed in a number of regional and national level targets. There is broad consensus, however, that in the absence of conservation action, biodiversity will continue to be lost at a rate unprecedented in the recent era. Remarkably, we lack a basic system to measure progress towards these targets and, in particular, we lack standard measures of biodiversity and procedures to construct and assess summary statistics. Here, we develop a simple classification of biodiversity indicators to assist their development and clarify purpose. We use European birds, as example taxa, to show how robust indicators can be constructed and how they can be interpreted. We have developed statistical methods to calculate supranational, multi-species indices using population data from national annual breeding bird surveys in Europe. Skilled volunteers using standardized field methods undertake data collection where methods and survey designs differ slightly across countries. Survey plots tend to be widely distributed at a national level, covering many bird species and habitats with reasonable representation. National species' indices are calculated using log linear regression, which allows for plot turnover. Supranational species' indices are constructed by combining the national species' indices weighted by national population sizes of each species. Supranational, multi-species indicators are calculated by averaging the resulting indices. We show that common farmland birds in Europe have declined steeply over the last two decades, whereas woodland birds have not. Evidence elsewhere shows that the main driver of farmland bird declines is increased agricultural intensification. We argue that the farmland bird indicator is a useful surrogate for trends in other elements of biodiversity in this habitat. PMID- 15814346 TI - The Living Planet Index: using species population time series to track trends in biodiversity. AB - The Living Planet Index was developed to measure the changing state of the world's biodiversity over time. It uses time-series data to calculate average rates of change in a large number of populations of terrestrial, freshwater and marine vertebrate species. The dataset contains about 3000 population time series for over 1100 species. Two methods of calculating the index are outlined: the chain method and a method based on linear modelling of log-transformed data. The dataset is analysed to compare the relative representation of biogeographic realms, ecoregional biomes, threat status and taxonomic groups among species contributing to the index. The two methods show very similar results: terrestrial species declined on average by 25% from 1970 to 2000. Birds and mammals are over represented in comparison with other vertebrate classes, and temperate species are over-represented compared with tropical species, but there is little difference in representation between threatened and non-threatened species. Some of the problems arising from over-representation are reduced by the way in which the index is calculated. It may be possible to reduce this further by post stratification and weighting, but new information would first need to be collected for data-poor classes, realms and biomes. PMID- 15814347 TI - Biodiversity trends in Europe: development and testing of a species trend indicator for evaluating progress towards the 2010 target. AB - This paper presents a trial of a species population trend indicator for evaluating progress towards the 2010 biodiversity target in Europe, using existing data. The indicator integrates trends on different species (groups), and can be aggregated across habitats and countries. Thus, the indicator can deliver both headline messages for high-level decision-making and detailed information for in-depth analysis, using data from different sources, collected with different methods. International non-governmental organizations mobilized data on over 2800 historical trends in national populations of birds, butterflies and mammals, for a total of 273 species. These were combined by habitat and biogeographical region to generate a pilot pan-European scale indicator. The trial indicator suggests a decline of species populations in nearly all habitats, the largest being in farmland, where species populations declined by an average of 23% between 1970 and 2000. The indicator is potentially useful for monitoring progress towards 2010 biodiversity targets, but constraints include: the limited sensitivity of the historical data, which leads to conservative estimates of species decline; a potential danger of ambiguity because increases in opportunistic species can mask the loss of other species; and failure to account for pre-1970 population declines. We recommend mobilizing additional existing data, particularly for plants and fishes, and elaborating further the criteria for compiling representative sets of species. For a frequent, reliable update of the indicator, sound, sensitive and harmonized biodiversity monitoring programmes are needed in all pan-European countries. PMID- 15814348 TI - Measuring marine fishes biodiversity: temporal changes in abundance, life history and demography. AB - Patterns in marine fishes biodiversity can be assessed by quantifying temporal variation in rate of population change, abundance, life history and demography concomitant with long-term reductions in abundance. Based on data for 178 populations (62 species) from four north-temperate oceanic regions (Northeast Atlantic and Pacific, Northwest Atlantic, North mid-Atlantic), 81% of the populations in decline prior to 1992 experienced reductions in their rate of loss thereafter; species whose rate of population decline accelerated after 1992 were predominantly top predators such as Atlantic cod (Gadus morhua), sole (Solea solea) and pelagic sharks. Combining population data across regions and species, marine fishes have declined 35% since 1978 and are currently less than 70% of recorded maxima; demersal species are generally at historic lows, pelagic species are generally stable or increasing in abundance. Declines by demersal species have been associated with substantive increases in pelagic species, a pattern consistent with the hypothesis that increases in the latter may be largely attributable to reduced predation mortality. There is a need to determine the consequences to population growth effected by the reductions in age and size at maturity, and in mean age and size of spawners, concomitant with population decline. We conclude that reductions in the rate of population decline, in the absence of targets for population increase, will be insufficient to effect a recovery of marine fishes biodiversity, and that great care must be exercised when interpreting multi-species patterns in abundance. Of fundamental importance is the need to explain the geographical, species-specific and habitat biases that pervade patterns of marine fishes recovery and biodiversity. PMID- 15814349 TI - Monitoring change in the abundance and distribution of insects using butterflies and other indicator groups. AB - Conservative estimates suggest that 50-90% of the existing insect species on Earth have still to be discovered, yet the named insects alone comprise more than half of all known species of organism. With such poor baseline knowledge, monitoring change in insect diversity poses a formidable challenge to scientists and most attempts to generalize involve large extrapolations from a few well studied taxa. Butterflies are often the only group for which accurate measures of change can be obtained. Four schemes, used successfully to assess change in British butterflies, that are increasingly being applied across the world are described: Red Data Books (RDB) list the best judgements of experts of the conservation status of species in their field of expertise; mapping schemes plot the changing distributions of species at scales of 1-100 km2; transect monitoring schemes generate time series of changes in abundance in sample populations of species on fixed sites across the UK; and occasional surveys measure the number, boundaries and size of all populations of a (usually RDB) species at intervals of 10-30 years. All schemes describe consistent patterns of change, but if they are to be more generally useful, it is important to understand how well butterflies are representative of other taxa. Comparisons with similarly measured changes in native bird and plant species suggest that butterflies have declined more rapidly that these other groups in Britain; it should soon be possible to test whether this pattern exists elsewhere. It is also demonstrated that extinction rates in British butterflies are similar to those in a range of other insect groups over 100 years once recording bias is accounted for, although probably lower than in aquatic or parasitic taxa. It is concluded that butterflies represent adequate indicators of change for many terrestrial insect groups, but recommended that similar schemes be extended to other popular groups, especially dragonflies, bumblebees, hoverflies and ants. Given institutional backing, similar projects could be employed internationally and standardized. Finally, a range of schemes designed to monitor change in communities of aquatic macro-invertebrates is described. Although designed to use invertebrates as a bio-indicator of water quality for human use, these programmes could be extended to monitor the 2010 biodiversity targets of the World Summit on Sustainable Development. PMID- 15814350 TI - Measuring the fate of plant diversity: towards a foundation for future monitoring and opportunities for urgent action. AB - Vascular plants are often considered to be among the better known large groups of organisms, but gaps in the available baseline data are extensive, and recent estimates of total known (described) seed plant species range from 200000 to 422000. Of these, global assessments of conservation status using International Union for the Conservation of Nature (IUCN) categories and criteria are available for only approximately 10000 species. In response to recommendations from the Conference of the Parties to the Convention on Biological Diversity to develop biodiversity indicators based on changes in the status of threatened species, and trends in the abundance and distribution of selected species, we examine how existing data, in combination with limited new data collection, can be used to maximum effect. We argue that future work should produce Red List Indices based on a representative subset of plant species so that the limited resources currently available are directed towards redressing taxonomic and geographical biases apparent in existing datasets. Sampling the data held in the world's major herbaria, in combination with Geographical Information Systems techniques, can produce preliminary conservation assessments and help to direct selective survey work using existing field networks to verify distributions and gather population data. Such data can also be used to backcast threats and potential distributions through time. We outline an approach that could result in: (i) preliminary assessments of the conservation status of tens of thousands of species not previously assessed, (ii) significant enhancements in the coverage and representation of plant species on the IUCN Red List, and (iii) repeat and/or retrospective assessments for a significant proportion of these. This would result in more robust Sampled Red List Indices that can be defended as more representative of plant diversity as a whole; and eventually, comprehensive assessments at species level for one or more major families of angiosperms. The combined results would allow scientifically defensible generalizations about the current status of plant diversity by 2010 as well as tentative comments on trends. Together with other efforts already underway, this approach would establish a firmer basis for ongoing monitoring of the status of plant diversity beyond 2010 and a basis for comparison with the trend data available for vertebrates. PMID- 15814351 TI - Tropical forest cover change in the 1990s and options for future monitoring. AB - Despite the importance of the world's humid tropical forests, our knowledge concerning their rates of change remains limited. Two recent programmes (FAO 2000 Forest Resources Assessment and TREES II), exploiting the global imaging capabilities of Earth observing satellites, have recently been completed to provide information on the dynamics of tropical forest cover. The results from these independent studies show a high degree of conformity and provide a good understanding of trends at the pan-tropical level. In 1990 there were some 1150 million ha of tropical rain forest with the area of the humid tropics deforested annually estimated at 5.8 million ha (approximately twice the size of Belgium). A further 2.3 million ha of humid forest is apparently degraded annually through fragmentation, logging and/or fires. In the sub-humid and dry tropics, annual deforestation of tropical moist deciduous and tropical dry forests comes to 2.2 and 0.7 million ha, respectively. Southeast Asia is the region where forests are under the highest pressure with an annual change rate of -0.8 to -0.9%. The annual area deforested in Latin America is large, but the relative rate (-0.4 to 0.5%) is lower, owing to the vast area covered by the remaining Amazonian forests. The humid forests of Africa are being converted at a similar rate to those of Latin America (-0.4 to -0.5% per year). During this period, secondary forests have also been established, through re-growth on abandoned land and forest plantations, but with different ecological, biophysical and economic characteristics compared with primary forests. These trends are significant in all regions, but the extent of new forest cover has proven difficult to establish. These results, as well as the lack of more detailed knowledge, clearly demonstrate the need to improve sound scientific evidence to support policy. The two projects provide useful guidance for future monitoring efforts in the context of multilateral environmental agreements and of international aid, trade and development partnerships. Methodologically, the use of high-resolution remote sensing in representative samples has been shown to be cost-effective. Close collaboration between tropical institutions and inter-governmental organizations proved to be a fruitful arrangement in the different projects. To properly assist decision-making, monitoring and assessments should primarily be addressed at the national level, which also corresponds to the ratification level of the multilateral environmental agreements. The Forest Resources Assessment 2000 deforestation statistics from countries are consistent with the satellite-based estimates in Asia and America, but are significantly different in Africa, highlighting the particular need for long-term capacity-building activities in this continent. PMID- 15814352 TI - Measuring coral reef decline through meta-analyses. AB - Coral reef ecosystems are in decline worldwide, owing to a variety of anthropogenic and natural causes. One of the most obvious signals of reef degradation is a reduction in live coral cover. Past and current rates of loss of coral are known for many individual reefs; however, until recently, no large scale estimate was available. In this paper, we show how meta-analysis can be used to integrate existing small-scale estimates of change in coral and macroalgal cover, derived from in situ surveys of reefs, to generate a robust assessment of long-term patterns of large-scale ecological change. Using a large dataset from Caribbean reefs, we examine the possible biases inherent in meta analytical studies and the sensitivity of the method to patchiness in data availability. Despite the fact that our meta-analysis included studies that used a variety of sampling methods, the regional estimate of change in coral cover we obtained is similar to that generated by a standardized survey programme that was implemented in 1991 in the Caribbean. We argue that for habitat types that are regularly and reasonably well surveyed in the course of ecological or conservation research, meta-analysis offers a cost-effective and rapid method for generating robust estimates of past and current states. PMID- 15814353 TI - Prospects for monitoring freshwater ecosystems towards the 2010 targets. AB - Human activities have severely affected the condition of freshwater ecosystems worldwide. Physical alteration, habitat loss, water withdrawal, pollution, overexploitation and the introduction of non-native species all contribute to the decline in freshwater species. Today, freshwater species are, in general, at higher risk of extinction than those in forests, grasslands and coastal ecosystems. For North America alone, the projected extinction rate for freshwater fauna is five times greater than that for terrestrial fauna--a rate comparable to the species loss in tropical rainforest. Because many of these extinctions go unseen, the level of assessment and knowledge of the status and trends of freshwater species are still very poor, with species going extinct before they are even taxonomically classified. Increasing human population growth and achieving the sustainable development targets set forth in 2002 will place even higher demands on the already stressed freshwater ecosystems, unless an integrated approach to managing water for people and ecosystems is implemented by a broad constituency. To inform and implement policies that support an integrated approach to water management, as well as to measure progress in halting the rapid decline in freshwater species, basin-level indicators describing the condition and threats to freshwater ecosystems and species are required. This paper discusses the extent and quality of data available on the number and size of populations of freshwater species, as well as the change in the extent and condition of natural freshwater habitats. The paper presents indicators that can be applied at multiple scales, highlighting the usefulness of using remote sensing and geographical information systems technologies to fill some of the existing information gaps. Finally, the paper includes an analysis of major data gaps and information needs with respect to freshwater species to measure progress towards the 2010 biodiversity targets. PMID- 15814354 TI - Background and interpretation of the 'Marine Trophic Index' as a measure of biodiversity. AB - Since the demonstration, in 1998, of the phenomenon now widely known as 'fishing down marine food webs', and the publication of a critical rejoinder by Food and Agricultural Organization (FAO) staff, a number of studies have been conducted in different parts of the world, based on more detailed data than the global FAO fisheries statistics originally used, which established the validity and ubiquity of this phenomenon. In this contribution, we briefly review how, rather than being an artefact of biased data, this phenomenon was in fact largely masked by such data, and is in fact more widespread than was initially anticipated. This is made visible here by comparing two global maps of trophic level (TL) changes from the early 1950s to the present. The first presents the 50-year difference of the grand mean TL values originally used to demonstrate the fishing down effect, while the second is based on means above a cut-off TL (here set at 3.25), thus eliminating the highly variable and abundant small pelagic fishes caught throughout the world. Based on this, we suggest that using mean TL as 'Marine Trophic Index' (MTI), as endorsed by the Convention on Biological Diversity, always be done with an explicitly stated cut-off TL (i.e. cutMTI), chosen (as is the case with our proposed value of 3.25) to emphasize changes in the relative abundance of the more threatened, high-TL fishes. We also point out the need to improve the taxonomic resolution, completeness and accuracy of the national and international fisheries catch data series upon which the cutMTI is to be based. PMID- 15814355 TI - Measuring conditions and trends in ecosystem services at multiple scales: the Southern African Millennium Ecosystem Assessment (SAfMA) experience. AB - The Southern African Millennium Ecosystem Assessment (SAfMA) evaluated the relationships between ecosystem services and human well-being at multiple scales, ranging from local through to sub-continental. Trends in ecosystem services (fresh water, food, fuel-wood, cultural and biodiversity) over the period 1990 2000 were mixed across scales. Freshwater resources appear strained across the continent with large numbers of people not securing adequate supplies, especially of good quality water. This translates to high infant mortality patterns across the region. In some areas, the use of water resources for irrigated agriculture and urban-industrial expansion is taking place at considerable cost to the quality and quantity of freshwater available to ecosystems and for domestic use. Staple cereal production across the region has increased but was outstripped by population growth while protein malnutrition is on the rise. The much-anticipated wood-fuel crisis on the subcontinent has not materialized but some areas are experiencing shortages while numerous others remain vulnerable. Cultural benefits of biodiversity are considerable, though hard to quantify or track over time. Biodiversity resources remain at reasonable levels, but are declining faster than reflected in species extinction rates and appear highly sensitive to land-use decisions. The SAfMA sub-global assessment provided an opportunity to experiment with innovative ways to assess ecosystem services including the use of supply demand surfaces, service sources and sink areas, priority areas for service provision, service 'hotspots' and trade-off assessments. PMID- 15814356 TI - Measuring the extent and effectiveness of protected areas as an indicator for meeting global biodiversity targets. AB - There are now over 100000 protected areas worldwide, covering over 12% of the Earth's land surface. These areas represent one of the most significant human resource use allocations on the planet. The importance of protected areas is reflected in their widely accepted role as an indicator for global targets and environmental assessments. However, measuring the number and extent of protected areas only provides a unidimensional indicator of political commitment to biodiversity conservation. Data on the geographic location and spatial extent of protected areas will not provide information on a key determinant for meeting global biodiversity targets: 'effectiveness' in conserving biodiversity. Although tools are being devised to assess management effectiveness, there is no globally accepted metric. Nevertheless, the numerical, spatial and geographic attributes of protected areas can be further enhanced by investigation of the biodiversity coverage of these protected areas, using species, habitats or biogeographic classifications. This paper reviews the current global extent of protected areas in terms of geopolitical and habitat coverage, and considers their value as a global indicator of conservation action or response. The paper discusses the role of the World Database on Protected Areas and collection and quality control issues, and identifies areas for improvement, including how conservation effectiveness indicators may be included in the database to improve the value of protected areas data as an indicator for meeting global biodiversity targets. PMID- 15814357 TI - Measuring biodiversity and sustainable management in forests and agricultural landscapes. AB - Most of the world's biodiversity will continue to exist outside protected areas and there are also managed lands within many protected areas. In the assessment of millennium targets, there is therefore a need for indicators to measure biodiversity and suitability of habitats for biodiversity both across the whole landscape/seascape and in specific managed habitats. The two predominant land uses in many inhabited areas are forestry and agriculture and these are examined. Many national-level criteria and indicator systems already exist that attempt to assess biodiversity in forests and the impacts of forest management, but there is generally less experience in measuring these values in agricultural landscapes. Existing systems are reviewed, both for their usefulness in providing indicators and to assess the extent to which they have been applied. This preliminary gap analysis is used in the development of a set of indicators suitable for measuring progress towards the conservation of biodiversity in managed forests and agriculture. The paper concludes with a draft set of indicators for discussion, with suggestions including proportion of land under sustainable management, amount of produce from such land, area of natural or high quality semi-natural land within landscapes under sustainable management and key indicator species. PMID- 15814358 TI - Turning science into policy: challenges and experiences from the science-policy interface. AB - This paper discusses key issues in the science-policy interface. It stresses the importance of linking the conservation and sustainable use of biodiversity to the Millennium Development Goals and to issues of immediate concern to policy-makers such as the economy, security and human health. It briefly discusses the process of decision-making and how the scientific and policy communities have successfully worked together on global environmental issues such as stratospheric ozone depletion and climate change, and the critical role of international assessments in providing the scientific basis for informed policy at the national and international level. The paper also discusses the drivers of global environmental change, the importance of constructing plausible futures, indicators of change, the biodiversity 2010 target and how environmental issues such as loss of biodiversity, stratospheric ozone depletion, land degradation, water pollution and climate change cannot be addressed in isolation because they are strongly interconnected and there are synergies and trade-offs among the policies, practices and technologies that are used to address these issues individually. PMID- 15814359 TI - Genetic alterations in brain tumors following 1,3-butadiene exposure in B6C3F1 mice. AB - The nervous system of the B6C3F1 mouse has rarely been a target for chemical carcinogenesis in the National Toxicology Program (NTP) bioassays. However, 6 malignant gliomas and 2 neuroblastomas were observed in B6C3F1 mice exposed to 625 ppm 1,3-butadiene (NTP technical reports 288 and 434). These mouse brain tumors were evaluated with regard to the profile of genetic alterations that are observed in human brain tumors. Alterations in the p53 tumor suppressor gene were common. Missense mutations were observed in 3/6 malignant gliomas and 2/2 neuroblastomas and were associated with loss of heterozygosity. Most of the mutations occurred in exons 5-8 of the p53 gene and were G-->A transitions, and did not involve CpG sites. Loss of heterozygosity at the Ink4a/Arf gene locus was observed in 5/5 malignant gliomas and 1/1 neuroblastoma, while the PTEN(phosphatase and tensin homologue) gene locus was unaffected by deletions. One of 2 neuroblastomas had a mutation in codon 61 of H-ras, while H-ras mutations were not observed in the malignant gliomas examined. Only 1 brain tumor has been reported from control mice of over 500 NTP studies. This malignant glioma showed no evidence of alterations in the p53 gene or K- and H-ras mutations. It is likely that the specific genetic alterations observed were induced or selected for by 1,3-butadiene treatment that contributed to the development of mouse brain tumors. The observed findings are similar in part to the genetic alterations reported in human brain tumors. PMID- 15814360 TI - Epididymal sperm granuloma induced by chronic administration of 2-methylimidazole in B6C3F1 mice. AB - Two-year mouse and rat bioassay studies of 2-methylimidazole (2-MI) conducted by the National Toxicology Program revealed that epididymal sperm granuloma(SG)s occurred only in male B6C3F1 mice in a dose-related manner. The present study characterized 2-MI-induced SGs in these epididymides. Groups of 50 male B6C3F1 mice were fed diets containing 0, 625, 1250, or 2500 ppm 2-MI for 105 weeks; the doses were equivalent to average daily doses of approximately 13, 40, or 130 mg/kg. Testes and epididymides were histopathologically reexamined. 2 Methylimidazole increased the incidence of epididymal SGs (0%, 0%, 6%, 12%, respectively). Histologically, most of the SGs exhibited rupture of epididymal ducts with focal aggregations of macrophages in interstitia. Lesions occurred in the proximal caput of the epididymis and/or efferent ducts, not in the corpus and cauda. In the testis, incidences of germinal epithelial atrophy (GEA) increased dose-relatedly (2%, 8%, 16%, 28%, respectively). All mice with epididymal SG developed testicular GEA. The grading scores of testicular GEA tended to be more severe in mice with SGs than those without. No epididymal SG or testicular GEA was observed in 6-month-interim-sacrificed mice. The results imply that 2-year treatment of B6C3F1 mice with 2-MI can induce epididymal SGs, primarily followed by more severe testicular GEA. The potential mechanism of SG induction by 2-MI is discussed. PMID- 15814361 TI - Ziracin-induced congenital urogenital malformations in female rats. AB - Spontaneous hypospadias is seldom observed in rats in contrast to its occurrence in 1 out of 250 human births. Ziracin, an antibacterial of the everninomycin class under development for serious enterococcal, staphylococcal, and streptococcal infections, caused anomalies of the external genitalia in F1 female rats and decreased reproductive performance. To characterize the urogenital malformations and determine the period of sensitivity to the effects of Ziracin during development, pregnant rats (F0) were administered 60 mg/kg IV of Ziracin from GD6 to LD21, GD6 to 13, GD14 to the last day of gestation or LD0 to 21. Controls received saline or placebo from GD6 to LD21. Ziracin-induced changes occurred in F1 rats exposed from GD6 to LD21 and GD14 to the last day of gestation, indicating that the period of sensitivity to Ziracin was from GD 14 to the last day of gestation. The urogenital abnormalities consisted of cranial displacement of the urethral opening within the vagina from its normal location at the tip of the genital tubercle. When the urethrovaginal junction occurred at the distal third of the vagina, it created an urogenital cloaca. As a result, ascending infections were seen in the urinary and genital tract. No differences in survivability, body weight, and date of vaginal opening were observed in F1 females. The estrous cycles were slightly prolonged. The mating and fertility indices were decreased as a result of the urogenital anomalies. The mammary glands of pregnant F1 females were underdeveloped, thus F2 pups from affected F1 females had a decreased survival rate. Although the cause of these effects is not known, the findings are consistent with a potential hormonal mechanism. PMID- 15814362 TI - Persistence of liver cirrhosis in association with proliferation of nonparenchymal cells and altered location of alpha-smooth muscle actin-positive cells. AB - This study was carried out to achieve pathological understanding for the persistence of cirrhosis induced by thioacetamide (TAA). Forty-five, male,21-day old, F344 rats were randomly allocated to group I and received drinking water as a control, and groups 2 and 3 given 0.015% or 0.03%TAA, respectively for 12 weeks. Two-third of animals per group were sacrificed, and remainder were maintained for a further 4 weeks without TAA treatment. Liver cirrhosis was induced in all animals in group 3 at week 12, with obvious increase of collagen content, and this persisted after cessation of TAA. Proliferating cell nuclear antigen (PCNA) positive labeling indices of nonparenchymal cells were increased significantly after cessation in groups 2 and 3 (p < 0.01). RT-RCR analysis of a smooth muscle actin (alpha-SMA) showed significant increase in group 3 compared to that of control at both time points (p < 0.05). Immunohistochemical staining of it demonstrated positive cells to mainly be located around regenerating hepatic nodules at week 12, however, they were focused into enlarged portal areas consisting of fibrous tissues and pseudo-bile ductular cells after the cessation. Taken together, we conclude persistence of liver cirrhosis could be associated with the proliferation of nonparenchymal cells and altered location of alpha-SMA positive cells. PMID- 15814363 TI - Induction of acute lung injury after intranasal administration of toxin botulinum a complex. AB - The inhalation of aerozolized botulinum toxin may represent a potential significant hazard to both military and civilian personnel. Since the lung is the primary target organ for inhaled toxin, the investigation reported herein was conducted to examine lung function in mice exposed to botulinum toxin A complex by intranasal route. Data includes lethality, symptomatology, measurement of respiratory function (minute ventilation, respiratory frequency, and tidal volume), and histopathology of the lungs. The clinical signs of intoxication are similar to those observed in foodborne botulism. Plethysmography revealed severe impairment of all respiratory parameters tested from 7 hours postexposure. Severe lung lesions, possibly secondary to the intoxication, were observed in mice who survived 14 days after the toxin challenge. These included intra-alveolar hemorrhage and interstitial edema. Mice immunized by the pentavalent (ABCDE) toxoid were protected against the neurotoxin (4 LD50) as revealed by the decrease of lethality and severity of nervous signs of intoxication, but not against histopathological changes in the lungs. These effects are nonspecific and require further experiments in order to specify the relationships between the pathology and the inflammatory process in the lung due to mediators such as cytokines,and possibly permanent physiological sequelae. PMID- 15814364 TI - The critical turn in clinical ethics and its continuous enhancement. AB - Taking the critical turn is one of the main tools of the humanities and inculcates an intellectual discipline that prevents ossification of thinking about issues and of organizational policies in clinical ethics. The articles in this "Clinical Ethics" number of the Journal take the critical turn with respect to cherished ways of thinking in Western clinical ethics, life extension, the clinical determination of death, physicians' duty to treat even at personal risk, clinical ethics at the interface of research ethics, and the pertinence of the Hippocratic Oath to clinical ethics. These articles challenge clinical ethicists to inculcate the intellectual discipline of the critical turn into everyday practice and continuous quality enhancement of clinical ethics. PMID- 15814365 TI - The nature of relative subjectivity: a reflexive mode of thought. AB - Ethical principles including autonomy, justice and equality function in the same paradigm of thought, that is, logocentrism--an epistemological predilection that relies on the analytic power of deciphering between binary oppositions. By studying observable behavior with an analytical approach, however, one immediately limits any recognition and possible understanding of modes of thought based on separate epistemologies. This article seeks to reveal an epistemological predilection that diverges from logocentrism yet continues to function as a fundamental component of ethical behavior. The issue of cancer disclosure in Japan is used to exemplify the nature of relative subjectivity (kankeiteki shukansei), a term I define and believe to be an epistemological predisposition fundamental to ethical behavior. Relative subjectivity denotes an epistemology quintessential to the behavior of individuals who attend to the respective tides of each particular situation, each interpersonal relationship or, in the context of clinical medicine, to the needs and values of each patient. PMID- 15814366 TI - Life-extension and the Malthusian Objection. AB - Dramatically extending the human lifespan seems increasingly possible. Many bioethicists object that life-extension will have Malthusian consequences as new Methuselahs accumulate, generation by generation. I argue for a Life-Years Response to the Malthusian Objection. If even a minority of each generation chooses life-extension, denying it to them deprives them of many years of extra life, and their total extra life-years are likely to exceed the total life-years of a majority who do not want life-extension. This is a greater harm to those who want extended life than the Malthusian harms to those who refuse extended life, both because losing an extra year of life is worse than enduring a year of Malthusian conditions, and because the would-be Methuselahs have more life-years at stake. Therefore, even if life-extension seems likely to cause severe overcrowding and resource shortages, that threat is not sufficient to justify society in restricting the development or availability of life-extension. PMID- 15814367 TI - Potentiality, irreversibility, and death. AB - There has been growing concern about whether individuals who satisfy neurological criteria for death or who become non-heart-beating organ donors are really dead. This concern has focused on the issue of the potential for recovery that these individuals may still have and whether their conditions are irreversible. In this article I examine the concepts of potentiality and irreversibility that have been invoked in the discussions of the definition of death and non-heart-beating organ donation. I initially focus on the recent challenge by D. Alan Shewmon to accepting any neurological criterion of death. I argue that Shewmon relies on a problematic and unrealistic concept of potentiality, and that a better, more realistic concept of potentiality is consistent with accepting a neurological criterion for death. I then turn to an analysis of how the concept of irreversibility has been used in discussion of non-heart-beating organ donation. Similarly, I argue that some participants in this discussion have invoked a problematic and unrealistic concept of irreversibility. I then propose an alternative, more realistic account of irreversibility that explains how "irreversibility" should be understood in the definition and criteria of death. PMID- 15814368 TI - In harm's way: AMA physicians and the duty to treat. AB - In June 2001, the American Medical Association (AMA) issued a revised and expanded version of the Principles of Medical Ethics (last published in 1980). In light of the new and more comprehensive document, the present essay is geared to consideration of a longstanding tension between physician's autonomy rights and societal obligations in the AMA Code. In particular, it will be argued that a duty to treat overrides AMA autonomy rights in social emergencies, even in cases that involve personal risk to physicians (e.g., bioterrorist attack, HIV infection, SARS). The argument will be made by way of the logic and language of the AMA Code through its history, commentaries, and precedents. It also will be shown that there are substantial reasons to believe that the logic of the Code is sound in morally relevant ways. The essay will conclude with some philosophical proposals suggesting a framework for the duty to render aid and the extension of those duties to physicians facing personal risks. PMID- 15814369 TI - Timing invitations to participate in clinical research: preliminary versus informed consent. AB - This article addresses the impact of the potential conflict between the roles of physicians who are both clinicians and researchers on the recruitment of persons into research trials. It has been proposed (1) that a physician breaches inter role confidentiality when he or she uses information gathered in his or her clinical role to inform patients about trials for which they may be eligible and (2) that clinician-researchers should adopt a model of preliminary consent to be approached about research prior to commencing a clinical relationship. This article argues that even if we grant the legitimacy of inter-role confidentiality (which is open to question), there are circumstances in which other obligations physicians bear override the obligation of inter-role confidentiality. Moreover, it is argued that the practice of preliminary consent is morally suspect and that such consent cannot be deemed valid. The article concludes with a series of recommendations of ways in which the legitimate concern regarding the conflicting roles of clinician-researchers can be addressed in the recruitment stage of research. PMID- 15814370 TI - The Hippocratic oath and contemporary medicine: dialectic between past ideals and present reality? AB - The Hippocratic Oath, the Hippocratic tradition, and Hippocratic ethics are widely invoked in the popular medical culture as conveying a direction to medical practice and the medical profession. This study critically addresses these invocations of Hippocratic guideposts, noting that reliance on the Hippocratic ethos and the Oath requires establishing (1) what the Oath meant to its author, its original community of reception, and generally for ancient medicine (2) what relationships contemporary invocations of the Oath and the tradition have to the original meaning of the Oath and its original reception (3) what continuity exists and under what circumstances over the last two-and-a-half millenniums of medical-moral reflections (4) what continuity there is in the meaning of professionalism from the time of Hippocrates to the 21st century, and (5) what social factors in particular have transformed the medical profession in particular countries. This article argues that the resources for a better understanding of medical professionalism lie not in the Hippocratic Oath, tradition, or ethos in and of themselves. Rather, it must be found in a philosophy of medicine that explores the values internal to medicine, thus providing a medical-moral philosophy so as to be able to resist the deformation of medical professionalism by bioethics, biopolitics, and governmental regulation. The Oath, as well as Stephen H. Miles' recent monograph, The Hippocratic Oath and the Ethics of Medicine, are employed as heuristics, so as to throw into better light the extent to which the Hippocratic Oath, tradition, and ethics can provide guidance and direction, as well as to show the necessity of taking seriously the need for a substantive philosophy of medicine. PMID- 15814371 TI - Impact of the effect of economic crisis and the targeted motorcycle safety programme on motorcycle-related accidents, injuries and fatalities in Malaysia. AB - In 1997, a Motorcycle Safety Programme (MSP) was introduced to address the motorcycle-related accident problem. The MSP was specifically targeted at motorcyclists. In addition to the MSP, the recent economic recession has significantly contributed to a reduction of traffic-related incidents. This paper examines the effects of the recent economic crisis and the MSP on motorcycle related accidents, casualties and fatalities in Malaysia. The autocorrelation integrated moving average model with transfer function was used to evaluate the overall effects of the interventions. The variables used in developing the model were gross domestic product and MSPs. The analysis found a 25% reduction in the number of motorcycle-related accidents, a 27% reduction in motorcycle casualties and a 38% reduction in motorcycle fatalities after the implementation of MSP. Findings indicate that the MSP has been one of the effective measures in reducing motorcycle safety problems in Malaysia. Apart from that, the performance of the country's economy was also found to be significant in explaining the number of motorcycle-related accidents, casualties and fatalities in Malaysia. PMID- 15814372 TI - An evaluation of the effectiveness of televised road safety messages in Ghana. AB - The goal was to evaluate the effectiveness of recent televised advertisements conducted by the National Road Safety Commission in Ghana. These concerned speeding and alcohol-impaired driving and were targeted towards commercial drivers. Focus group discussions were conducted with 50 commercial drivers in four cities. Discussions addressed coverage, clarity and appropriateness of messages, including suggestions for improvements. Most discussants indicated that the messages were clear and appropriate. Television reached all participants in this urban group. However, they felt that other modes of communication, such as flyers and radio, should also be used to reach drivers who did not own televisions. A particular problem was language. The advertisements had been in English and Akan (the most common vernacular language). Participants wanted the messages diversified into more of the major Ghanaian languages. Some participants were unclear on the behaviour that the advertisements were telling viewers to take. Participants advocated greater involvement by police in road safety and called for laws banning the sale of alcohol at bus stations. The advertisements reached and were understood by most of the target audience. Opportunities for strengthening the messages included using other media; increasing the number of languages; and stressing the change in behaviour being recommended. Overall road safety activities would be strengthened by increasing accompanying law enforcement activities related to speed and alcohol-impaired driving. To the authors' knowledge this is the first formal evaluation of a road safety social marketing programme in a low-income sub-Saharan African country. This evaluation will hopefully assist Ghana and other similar countries in strengthening road safety work. PMID- 15814373 TI - Does poverty lead to non-fatal unintentional injuries in rural Vietnam? AB - The aim of this study was to identify associations between poverty at the household level and unintentional injury morbidity. A cohort consisting of 24,874 person-time episodes, representing 24,776 people living in 5,801 households (classified into rich, middle income and poor by local authorities in 1999) was followed during 2000, in order to identify and assess non-fatal unintentional injuries. Incidence rate ratios were calculated using a Poisson regression model. The results showed that poverty was a risk factor for unintentional injuries generally. When looking at different types of injury, poverty was a risk for home, work and "other" injuries, protective for school injuries, while the risk of traffic injuries was not affected. The results also showed that communes in mountainous areas were at higher risk for home, work and other injuries. Overall, poverty was associated with unintentional injury morbidity. However, the relationship varied by sex, age and type of injury. Specifically, poverty increased the risk for children and elderly people being injured at home, and for adults (15-59 years) being affected by work injuries. PMID- 15814374 TI - Inflatable rescue boat-related injuries in Queensland surf lifesavers: the epidemiology - biomechanics interface. AB - The objective was to describe the relationship between epidemiological and biomechanical factors in the causal pathway of inflatable rescue boat (IRB) related injuries in Australian surf lifesavers; to develop epidemiological and biomechanical methodologies and measurement instruments that identify and measure the risk factors, for use in future epidemiological studies. Epidemiological and biomechanical models of injury causation were combined. Host, agent and environmental factors that influenced total available force for transfer to host were specified. Measurement instruments for each of the specified risk factors were developed. Instruments were piloted in a volunteer sample of surf lifesavers. Participant characteristics were recorded using demographic questionnaires; IRB operating techniques were recorded using a custom-made on board camera (Grand RF-Guard) and images of operating techniques were coded by two independent observers. Ground reaction forces transmitted to the host through the lifesaver's feet at the time of wave impact were measured using a custom built piezoelectric force platform. The demographic questionnaire was found practical; the on-board camera functioned successfully within the target environment. Agreement between independent coders of IRB operating technique images was significant (p < 0.001) with Kappa values ranging from 0.5 to 0.7. Biomechanical instruments performed successfully in the target environment. Peak biomechanical forces were 415.6N (left foot) and 252.9N (right foot). This study defines the relationship between epidemiological and biomechanical factors in modifying the risk of IRB-related injury in a population of surf lifesavers. Preliminary feasibility of combining epidemiological and biomechanical information has been demonstrated. Further testing of the proposed model and measurement instruments is required. PMID- 15814375 TI - The neglected epidemic: road traffic accidents in a developing country, State of Qatar. AB - The objective of the study is to explore the pattern of road traffic accidents (RTA) and their causes in the State of Qatar. A total of 52,160 RTA, 1130 injuries and 85 fatalities were registered during the year 2000. The data on RTA, injuries and fatalities was obtained from the Traffic Department of the Ministry of Interior, Supreme Council for Planning and Ministry of Public Health. The major cause of traffic accidents in Qatar was careless driving (71%). Relatively there was a more rapid increase in the number of registered vehicles (155%), but accidents were only (61%) in 2000 compared to the year 1983. The majority of victims (53%) were in the age group (10-40) "the most productive class in Society". Forty-three per cent of the total drivers who died due to RTA were in the age group (10-19) who were unlicensed drivers. In 2000, deaths due to RTA were the third leading cause of death after the diseases of the circulatory system and cancer. In conclusion, it is possible to control the epidemic of road traffic injuries through strict policy interventions, mass media and a national traffic campaign to increase the use of seat belts. PMID- 15814376 TI - Causes and consequences of moped injuries in children and adolescents. PMID- 15814377 TI - Medium-term physical and behavioural sequelae of motor vehicle occupant injuries in children. PMID- 15814378 TI - Scope and patterns of tourist injuries in the European Union. AB - European Union (EU) countries are among the leading tourist destinations in the world. Despite growing consumer demands on the safety of travelling, injuries amongst tourists remain an essentially invisible problem. Unique national and regional data sources are the only means by which the important negative impact of injuries on the health of non-domestic tourists in the EU-15 is underlined. With an estimated number of 3800 tourists dying each year, injuries account for up to 30% of fatalities during vacation. This toll reflects an increased risk of mortality in tourists compared to the domestic population. Amendments to national and EU health and injury monitoring are suggested in order to enable authorities to examine personal risks to travellers in more detail. PMID- 15814379 TI - The in situ performance of playground surfacing: implications for maintenance and injury prevention. PMID- 15814380 TI - Establishing site specific reference levels for fungi in outdoor air for building evaluation. AB - Culturable airborne fungal spore sampling at five building sites during 2002-2003 provided a bank of outdoor data (102 samples total) to evaluate differences in levels of individual species of airborne fungi during the morning and afternoon hours. A minimum of 15 (outdoor) air samples was collected at each site, and data were segregated into morning (before noon) and afternoon subsets. Significant differences in airborne levels for all detected fungal types between the morning and afternoon subsets were determined for each site, using a direct calculation of probability. Significance was defined by differences in frequency of detection above the combined median (p=0.90 or greater) for the respective fungal type. The levels of various species of fungi in the outdoor air varied significantly between morning and afternoon data sets at all five sites, with no pattern by species, time of day, or location. Levels of Penicillium, Aspergillus, or other fungal species associated with problematic buildings if detected outdoors, can be significantly greater in the morning or afternoon (or exhibit no significant difference) on any given day. The data does not indicate laboratory analysis as a major contributor to the variability exhibited in bioaerosols, and underscores the necessity of collecting sufficient number of samples in the outdoor air in both the morning and afternoon to prevent bias when comparing a suspect indoor environment to outdoor conditions. PMID- 15814381 TI - A survey of private sector respirator use in the United States: an overview of findings. AB - Limitations of previous surveys of respirator use led the National Institute for Occupational Safety and Health (NIOSH) and the Bureau of Labor Statistics to undertake a survey of respirator use and practices among U.S. private sector employers. The survey was mailed to 40,002 private sector establishments in August 2001; the responses were used to develop national estimates. Respirator use was required in 4.5% of establishments and for 3.1% of employees. Of the establishments requiring respirator use, 95% used air-purifying respirators and 17% used air-supplied respirators. Manufacturing; mining (including oil and gas extraction); construction; and agriculture, forestry, and fishing had the highest rates of establishment respirator use. Respirators were used most frequently to protect against dust/mist, paint vapors, and solvents. Large percentages of establishments requiring respirator use had indicators of potentially inadequate respirator programs. Of establishments requiring respirator use, 91% had at least one indicator of a potentially inadequate respiratory protection program, while 54% had at least five indicators. The survey findings suggest that large numbers of employers may not follow NIOSH recommendations and Occupational Safety and Health Administration (OSHA) and Mine Safety and Health Administration (MSHA) requirements for the selection and use of respirators, potentially putting workers at risk. The findings will aid efforts to increase the appropriate use of respirators in the workplace. PMID- 15814382 TI - Brachial plexus injuries and shoulder dystocia: medico-legal commentary and implications. AB - Brachial plexus injuries may occur in association with or without recognised shoulder dystocia. Until recently many medico-legal experts argued that these injuries resulted from the negligent use of excessive or misdirected traction. These experts did not accept that the natural forces of labour could cause the injury. In cases of recognised shoulder dystocia, modern obstetric practice has recognised the importance of correct maternal positioning and the need to avoid excessive or misdirected traction, also the importance of not applying a rotational force to the fetal head. Informed practice and the increased use of caesarean section has not reduced the number of brachial plexus injuries. Judges in two recent trials found that the natural forces of labour were the cause of the injury and that the birth assistants had not caused the damage. This has important implications for medical note writing, teachers and risk managers, medico-legal experts and lawyers. PMID- 15814383 TI - Pregnancy following endometrial ablation: a review article. AB - This review summarises the 70 pregnancies reported following endometrial ablation by various techniques, including a those in a case report from the authors' own experience. Thirty-one viable pregnancies resulted, with a high proportion of complications. These included a perinatal mortality rate of 12.9% (4 cases), and an overall prematurity rate of 42%, which reduced to 31% after excluding iatrogenic deliveries performed for other complications. Eight (26%) cases had a morbidly adherent placenta and a further two cases required a manual removal of placenta. Seven (39%) of the term births were reported as having malpresentations and 71% of the total births were by caesarean section. The complications are discussed and a comparison is made with Asherman's Syndrome. In addition, the various factors that may be relevant to fertility after endometrial ablation are discussed. PMID- 15814384 TI - Reversal of the decision for caesarean section in the second stage of labour on the basis of consultant vaginal assessment. AB - During a 5-year period there were 32 cases where the vaginal assessment performed by a specialist registrar in the second stage of labour was re-assessed within 15 minutes by a consultant obstetrician. The examination was prompted by a request for permission to perform a caesarean section in the second stage of labour. The results suggest a significant discrepancy between the consultants and the specialist registrar's findings, with 44% of the cases indicating a difference in the position of the head, and 81% a difference in the station of the head. No comment was made about caput or moulding in the majority of cases (94%). The study findings suggest that vaginal examination, like instrumental delivery, is a skill that is being eroded and will require formal instruction to address this problem. PMID- 15814385 TI - Term preeclampsia is associated with minimal histopathological placental features regardless of clinical severity. AB - Preeclampsia (PET) is a serious complication of pregnancy, which is associated with uteroplacental disease and reduced uteroplacental perfusion. One of the histological features in placentas from pregnancies complicated by PET is infarction, representing focal severe uteroplacental ischaemia. This study examines the relationship between gestation at induced delivery and the prevalence of placental infarction using a placental pathology database to identify induced or operative deliveries on the basis of severe PET. The clinical and pathological findings were reviewed. Thirty-seven cases were identified, (4.9% of all placentas submitted). In 16 (43%), non-peripheral significant infarcts were identified histologically, including 13/20 (65%) requiring delivery before 34 weeks' compared to 3/17 (17%) requiring delivery > or = 34 weeks (z=2.9, P<0.01). Histological infarction is common in placentas from pregnancies complicated by severe PET but the prevalence is significantly greater in cases requiring delivery at earlier gestations, even when similar clinical indications for delivery are applied. PMID- 15814386 TI - Outcome of fetal pyelectasis diagnosed antenatally. AB - Renal pelvis dilatation is a common anomaly detected at the 20-week anomaly scan. The aim of this study was to evaluate the postnatal outcome of fetal pyelectasis in an unselected obstetric population. This was a retrospective study of fetal pyelectasis detected at midtrimester anomaly scan. Fetal pyelectasis was classified as mild if the anteroposterior renal pelvis diameter measured >5 mm-< or= 7 mm, moderate >7-< or = 10 mm and severe as >10 mm. Persistant fetal pyelectasis was defined as >7 mm in the third trimester. RESULTS: During the study period fetal pyelectasis was identified in 74 women. 6 were excluded, as case records were not available. Mild pyelectasis was identified in 38/67(56.7%), moderate pyelectasis in 20/67(29.8%) and severe pyelectasis in 9/67(13.4%). None of the infants in the mild pyelectasis subgroup required surgery. 3/29(10%) in the moderate and severe pyelectasis subgroup required postnatal surgery. Of the 38 with mild fetal pyelectasis 13.1% had a positive postnatal diagnosis out of which 7.9% had vesico-ureteric reflux, 2.6% renal dysplasia and 2.6% pelvi ureteric junction (PUJ) stenoses. CONCLUSION: 74% of pregnancies in the mild pyelectasis subgroup demonstrated spontaneous resolution. However persistent mild pyelectasis may lead to postnatal morbidity and should be followed up. PMID- 15814387 TI - How useful is the Helen Keller food frequency chart in the determination of the vitamin A status in pregnancy? AB - Vitamin A deficiency sub clinical or overt, is associated with adverse maternal, fetal and neonatal outcome. This is also true for an excess of vitamin A. The challenge in pregnancy is to detect sub clinical vitamin A deficiency in patients for whom supplements or dietary manipulation will be of benefit. This was a cross sectional case controlled study at the University of Benin Teaching Hospital to compare the Helen Keller Food Frequency Chart with biochemical methods in the determination of vitamin A status in pregnancy. Data was collected from Antenatal patients (142). Using serum Biochemistry three categories of patient were recognized. Patients with normal vitamin A levels (N=100 women with blood vitamin A within two standard deviation of the mean) Twenty-four women (24) had low vitamin A levels (N=24, patients with blood vitamin A level at less than 2 standard deviation below the mean). Eighteen patients (18) had high vitamin A levels (patients with blood vitamin A levels at greater than two standard deviation above the mean). All recruited patients had a dietary assessment using the Helen Keller Food Frequency Chart. The Helen Keller Food Frequency Chart (HKFFC) was found to have a high degree of sensitivity (74.5%) and a high specificity (75%) in detection of patients with vitamin A deficiency. The positive predictive value was 93.62%. The low negative predictive rate of 37.5% however implies that a positive test is more important than a negative test. The HKFFC was unable to differentiate patients with normal or high vitamin A levels. Dietary assessment with the HKFFC is a cheap effective method to detect sub clinical vitamin A deficiency in pregnancy. It is an easy cost effective screening tool to select patients for whom dietary manipulation and or vitamin A supplementation may be beneficial. PMID- 15814388 TI - Familial partial lipodystrophy complicated by pre-eclampsia. PMID- 15814389 TI - Determinants of pain perception in labour among parturients at the University College Hospital, Ibadan. AB - This cross-sectional study of one thousand parturients aims to evaluate the factors, which are associated with pain perception in labour and to identify women who will benefit most from analgesia in labour. The instrument applied was a structured questionnaire incorporating the Box numerical scale (BNS) for pain assessment. Patients who delivered by elective or emergency caesarean section and women who did not give informed consent were excluded from the study. Spearman's and Pearson's correlational analyses were applied. Pain scores showed significant correlation with age (r=- 0.087, p<0.01), parity (r=- 0.226, p<0.01), gestational age at delivery (r=- 0.074, p<0.05), onset of labour (rho=0.195, p<0.01), mode of delivery (rho=0.160, p<0.01), booking status (rho=- 0.070, p<0.05) and educational status (rho=0.182, p<0.01). Certain groups of patients would benefit from obstetric analgesia. These patients include, nulliparous patients, young patients, patients who have had labour induced, those with preterm deliveries and those with an assisted vaginal delivery especially if they are well educated. PMID- 15814390 TI - The value of routine haemoglobin concentration measurement before caesarean section. AB - Women undergoing caesarean section routinely have a haemoglobin concentration check before delivery. We compared the haemoglobin concentration of 311 women taken at their 32-?34 week antenatal visit with their preoperative level. A significant rise from 11.04 g/dL to 11.51 g/dL was seen (mean (95% CI) 0.47 (0.37 0.57 g/dL)). This increase was most marked in the 201 women having emergency procedures, and there was a significant negative correlation between the 32 and 34 weeks level and the net change in haemoglobin concentration (r=-0.366 (P<0.001)). From our results we suggest that in women with an otherwise uncomplicated pregnancy, and a satisfactory haemoglobin concentration at 32-34 weeks, a repeat estimation of the blood count is unnecessary before operative delivery. PMID- 15814391 TI - Pre-induction cervical ripening: transcervical foley catheter versus intravaginal misoprostol. AB - The object of this study was to compare the effectiveness of the intravaginal Misoprostol and transcervical Foley catheters as pre-induction cervical ripening agents, to estimate the proportion of patients achieving vaginal delivery and to compare the complications of labour and foetal outcome between the two groups. The study was a prospective, randomised study of pregnant women, with singleton pregnancies who presented for antenatal care and delivery at the University College Hospital (UCH), Ibadan, Nigeria. Ninety-nine patients were invited to participate and ninety-six (96) agreed. No patient withdrew from the study. The patients were assigned by means of computer-generated random numbers to receive transcervical Foley catheters (Size 16F, with 30 ml balloon capacity) or 50 microg intravaginal Misoprostol (Cytotec tablet, Searle & Co., Chicago). Fifty (50) patients received intravaginal Misoprostol and Forty-six (46) received Transcervical Foley catheters. The proportions of nulliparous, primiparous and multiparous patients were 52, 20 and 28% in the misoprostol group and 43.5, 26.1 and 30.4%, respectively, in the Foley catheter group. The time to achieve a favourable cervical status was significantly shorter in the Misoprostol group, with 98.0% of the subjects attaining Bishop score > or = 6 within 6-12 hours of insertion of the study agent, in contrast to 69.0% of the subjects in the Foley catheters group (P<0.001). Thirteen (26.6%) and three (6.5%) patients in the Misoprostol and Foley catheters groups, respectively, went into labour while undergoing cervical ripening and all had uneventful vaginal deliveries (P<0.05). The induction-delivery interval did not differ significantly between the groups. The incidence of caesarean delivery was 6.0% in the Misoprostol group compared with 2.2% in the Foley catheter group (P=0.62). Instrumental vaginal delivery rates were similar in both groups. Overall, the mode of delivery did not differ significantly between the groups. The number of neonates with 1-minute Apgar score <7 did not differ significantly in both groups and no neonate had 5-minute Apgar score <7. Meconium stained liquor was noticed in 5 (Misoprostol) vs 2 (Foley catheters) patients in labour. None of the neonates had any features suggestive of meconium aspiration. Labour complications were mainly precipitate labour {2 (Misoprostol) vs 1 (Foley catheters) } and 1 patient with transient tachysystole (> or =6 contractions in 10 minutes for two consecutive 10-minute periods) in the Misoprostol group. Hyperstimulation was not noticed in any of the patients in either arm of the study groups. Intravaginal Misoprostol is as effective a pre-induction cervical ripening agent as transcervical Foley catheters, with added advantages of shorter duration of cervical ripening, reduced oxytocin requirement for induction of labour and greater acceptability to patients. The incidence of caesarean sections, other labour complications and the foetal outcome were similar with both methods. PMID- 15814392 TI - Reduction in maternal mortality due to sepsis. AB - The present study was undertaken at a rural medical institute in India to analyse the trends in maternal mortality due to sepsis and the factors associated with change, if any. During the study period of 20 years, a total of 37,155 women delivered, 192 deaths occurred and forty deaths (20.83%) were due to sepsis and it's sequlae. It was revealed that there is a definite decrease in the proportion of deaths due to sepsis, to 10% in the last five years from 35% in earlier years. The change seems to be due to the advocacy of clean deliveries and reduction in case fatality because of alterations in medication and earlier surgical intervention. However the percentage contribution of septic abortion has remained the same. Septic abortion continues to exist inspite of all the current laws and discussion about the availability of a liberal law, which permits abortion almost on request. Most of the women who had died due to septic abortion were married (65%). Deaths due to septic abortion, are persisting even in married women and it is a matter of concern for health providers, policy makers and governments. PMID- 15814393 TI - A worldwide review of the uses of the uterine compression suture techniques as alternative to hysterectomy in the management of severe post-partum haemorrhage. AB - Postpartum haemorrhage (PPH) is a worldwide problem. The historical background dates back to William Smelley's in the seventeenth century in his famous treaty of the theory and practice of midwifery in 1752. Changes in clinical factors and surgical expertise compel the modern day midwife and obstetrician to be vigilant in identifying risk factors and apply appropriate solution early. The recent confidential enquiry into maternal death (why mothers die (2000-2002)) identifies areas of substandard care. The rising caesarean section rate adds to the rising incidence of PPH. The reduction in junior doctor's hours may limit the pool of experienced obstetric surgeons available to manage severe PPH competently. There can be major complications following radical surgery for PPH. These include loss of fertility, other morbidity and even maternal death. The invention of the B Lynch surgical technique for the conservative management of PPH was first performed and reported by a consultant obstetrician and gynaecological surgeon in Milton Keynes NHS Trust publishing the first series of cases in BJOG 1997. This has made a significant impact on the conservative surgical management of massive PPH. There are now over 1300 successful applications of this technique worldwide (CB-Lynch personal communication). Other similar or modified techniques such as Cho's Square Suture and Haymen's modification of the B-Lynch Suture Technique have been introduced adding to more available methods of conservative surgery. The current list of publications of successful application of the B-Lynch compression technique is encouraging and more outcome data can be reported by a letter or e-mail to enquiries@cblynch.com. Obstetricians and midwives both in developed and underdeveloped countries should seek training and attend fire drills in PPH control to avoid maternal morbidity and death. There should be special concentration on effective conservative surgery such as uterine compression techniques to avoid major morbidity and loss of fertility. PMID- 15814394 TI - Uterine compression suture without hysterotomy--why a non-absorbable suture should be avoided. AB - We describe 2 cases of uterine compression suture without hysterotomy, only described once in the literature previously (Hayman et al. 2002). We consider in detail the suture material used for this technique and show photos of the compression suture at laparoscopy 4 weeks after insertion to demonstrate why it is inappropriate to use a non-absorbable suture. Modified compression sutures are being used increasingly and a wide variety of suture materials are being chosen, including vicryl, PDS and nylon (verbal communications). We feel it important to report our findings so that others can avoid the use of non or slowly absorbable sutures. PMID- 15814395 TI - One stop fertility clinic. AB - The objective of this study was to determine the feasibility and acceptability of out-patient based investigation of infertile couples using a prospective observational study based in a large undergraduate teaching hospital. We studied couples referred to secondary care for investigation of their infertility. Investigations involved pelvic ultrasound, diagnostic hysteroscopy and culdoscopy. The main outcome measures were feasibility of investigations, findings, patient views, and a management plan. RESULTS: 199 of 347 (57.3%) couples referred met our selection criteria, and 162 of this group have attended. Thirty-one (19.1%) were judged to be unsuitable for culdoscopy, and culdoscopy failed in a further 29 (17.9%). The average time for the three procedures was 41.2 (SD 17.2) minutes, and over 1/3 of patients were found to have pelvic pathology. The investigations were well tolerated and there was only one complication necessitating admission to hospital. Most patients appreciated the need for a single hospital visit and the availability of immediate results. CONCLUSIONS: A One Stop approach to the investigation of infertility is attractive but not suitable for or desired by all infertile couples. Organisational issues were also identified. PMID- 15814396 TI - The cost of out-patient culdoscopy compared to in-patient laparoscopy in women with infertility. AB - Diagnostic laparoscopy is a common procedure performed in many hospitals worldwide to investigate infertile women. However, morbidity and cost prevent it being considered a first-line diagnostic tool. If out-patient culdoscopy could replace in-patient laparoscopy then a major component of the cost of investigations could be avoided. We studied ten consecutive infertile patients who had laparoscopy under general anaesthesia and another ten patients who had culdoscopy under local anaesthesia in The One Stop Fertility Clinic (OSFC) in The Royal Free Hospital in London. The costs for each patient from both groups was calculated and compared. We found that out-patient investigation in an OSFC produced a saving of over 350 pounds sterling per case or 28% to the hospital compared with in-patient investigation. Although there were other factors which may influence the costs of out-patient culdoscopy, it did provide noticeable savings to the healthcare system. PMID- 15814397 TI - Role of rescue IVF-ET treatment in the management of high response in stimulated IUI cycles. AB - Rescue in-vitro fertilisation and embryo transfer (IVF-ET) has been used in high response gonadotrophin intrauterine insemination (IUI) cycles to minimise the risks of ovarian hyperstimulation and multiple gestation. Such unplanned IVF treatment increases the cost of treatment. But can this added cost and the risks associated with IVF be justified? We present our experience with this treatment using clinical pregnancy and live birth rates as the primary outcomes. Between 1998 to 2001, 40 women undergoing IUI cycles who over responded (>3 follicles measuring >15 mm in diameter on the planned day of hCG administration) to gonadotrophin were offered the choice of conversion to IVF-ET or cancel the cycle. 17/40 declined rescue IVF/ET and had their cycles cancelled. 23/40 converted to IVF/ET and underwent transvaginal oocyte retrieval. 21/23 had embryo transferred. The clinical pregnancy and live birth rates were 52% and 48%, respectively. Rescue IVF-ET offers excellent clinical pregnancy and live birth rates in high responders. However, affordability can be an obstacle in the utilization of this treatment option. PMID- 15814398 TI - Follicle tracking of women receiving clomiphene citrate for ovulation induction. AB - Forty-five cycles of forty subfertile women were monitored with follicle tracking. Thirty women were anovulatory and 10 had unexplained infertility. Thirty women responded to the initial dose of Clomiphene Citrate (CC) and 10 remained anovulatory. Of those who ovulated with the first dose of CC, 25 developed one follicle, 2 developed 2 follicles and 3 developed three follicles. All the women that produced three follicles belonged in the unexplained infertility group. Eight women from the anovulatory group developed 1 follicle, two developed 2 follicles and 8 failed to respond. Provision of universal follicle tracking proved to be time consuming and required significant resources. Our results indicate that if there is a place for selective monitoring that should include women with unexplained infertility. We should also offer the first scan at around day 12 of the cycle as this could detect multiple follicle development. Patients with Polycystic Ovaries that start on 50 mgs of CC may not necessarily need follicle tracking as they usually do not respond to that dose. PMID- 15814400 TI - Sexual behaviour of secondary school adolescents in Ilesa, Nigeria: implications for the spread of STIs including HIV/AIDS. AB - This research examined the sexual behaviour and perception of the risk of HIV/AIDS and other sexually transmitted infections (STIs) among adolescents in Nigerian secondary schools. Quantitative data was collected from 450 senior secondary school 1 and 2 students who were selected from 5 of the 22 co educational secondary schools in Ilesa using a multistage sampling technique. In addition qualitative data was collected by focus group discussion (FGD) with 8 groups (4 for each of the sexes) of ten randomly selected respondents from 4 other secondary schools. Sixty three per cent had had sexual intercourse. There was no statistically significant (p>0.05) difference between the genders [females (52.3%) males (77.8%)], although males appeared to be more sexually active. The median age at first intercourse was 12 years with a range of 6-19 years. Many of the respondents had multiple sexual partners. There is poor perception of the risk of sexually transmitted infections (STIs) including that of human immune deficiency virus (HIV). It was concluded that there is a high risk of the spread of STIs, including HIV/AIDS in the study population. In view of the high prevalence of sexual intercourse and the reproductive health problems associated with STI, HIV/AIDS, it is recommended that a structured family life education (FLE) curriculum should be instituted for all schools starting at an early age. PMID- 15814399 TI - Comparison of carbon dioxide and room air pneumoperitoneum for day-case diagnostic laparoscopy. AB - Laparoscopic examination is useful in the evaluation of infertile women. To perform this test, pneumoperitoneum is required to distend the abdomen, improve visibility and displace the intestines out of the pelvis. Several gases have been used to achieve this purpose including nitrous oxide (N2O), carbon dioxide (CO2), helium, xenon and air. This is a prospective study in a private fertility centre comparing CO2 and room air pneumoperitoneum for diagnostic laparoscopy. Also the safety of room air was evaluated. One hundred and forty five patients received CO2 (group 1, n=73) or room air (group 2, n=72) pneumoperitoneum. Ketamine anaesthesia was used for all the patients and the cost of the procedure was same for the study. CO2 pneumoperitoneum offered better visibility during laparoscopy and the patients also had a better outcome: Wound infection (CO2-2/73: 2.7% Vs. Air-11/72: 15.3%); abdominal discomfort (feeling of retained gas in the abdomen) CO2-5/73: 6.9% Vs. Air-61/72: 84.7%) and shoulder pain (CO2-0/73; 0.0% Vs. Air 56/72: 77.8%). Group 1 patients returned to normal activity earlier (1.5 days +/- 1.3 SD Vs. 4.8 days +/- 2.1 SD). We conclude that CO2 pneumoperitoneum has a better outcome than Air especially for day-case diagnostic laparoscopy. However, room Air pneumoperitoneum is safe, cheap, and available and may be recommended for low resource settings. PMID- 15814401 TI - Assessment of contraceptive services in a maternity unit of a district general hospital in the UK. AB - A survey was undertaken to assess the quality of contraceptive services provided to women delivering in a District General Hospital. Separate questionnaires were given to the women and the midwives giving the contraceptive advice. Response rate of the survey was 87%. Thirty eight per cent of women reported having some discussion on contraception during antenatal period. All the women had received contraceptive advises during the postpartum period. Eighty one per cent of women felt that sufficient information and enough time was given to discuss contraception. However analysis of questionnaires indicated that most received information only on short-term contraceptive methods. Out of 64 midwives surveyed, 52 had attended a family planning course. However 37 of them had attended the course at least 3 years earlier. Thirty eight midwives felt that they have adequate knowledge to give advice but all of them reported that they should receive further training. There should be provision for comprehensive discussion regarding contraceptive options during both antenatal and postpartum periods. Midwives require regular training in family planning. PMID- 15814402 TI - Contraceptive prevalence among young women in Nigeria. AB - In almost all societies in the world there is an increase in sexual activity among young persons (10-24 years). There is a need for contraceptive use among them to prevent high risk unintended pregnancies. This study was carried out to determine the contraceptive prevalence among young women in Nigeria. The study population was 832 young women between the ages of 15 and 24 years. Only 11.1% of the respondents had ever used contraceptives although 63.2% of them had had sexual intercourse. Contraceptive usage was significantly higher (p<0.05) among the single sexually active women (38.5%) than among the married women (7.7%). Only 7.3% of respondents are currently using a contraceptive method. The main methods in use were condoms and pills. There is an unmet need for contraceptives among young women in Nigeria. It is necessary to improve these young women's abilities to protect themselves against unwanted pregnancies. PMID- 15814403 TI - The accuracy of Bardscan: a new tool for the measurement of the bladder volume. AB - The Bardscan has been recently introduced into clinical practice with the potential advantage of providing real time ultrasound imaging and hence the ability for an accurate estimation of bladder volume, and concurrently the detection of any incidental bladder pathology. Independent clinical data regarding this machine is limited, and therefore this study was carried out to ascertain whether the Bardscan machine is accurate in the estimation of bladder volume for women in the urodynamic clinic and in the postoperative period. Fifty four women had a Bardscan estimation of their bladder volume which was compared with an immediately obtained catheter volume. The results show that the Bardscan volumes correlate closely with the bladder volumes detected on catheterization (r =0.982, p=0.16). This study has shown that the Bardscan is an accurate tool in the measurement of bladder volume in women with the advantages of being safe, non invasive and well tolerated by patients. The aim of this study was to evaluate the accuracy of the Bardscan machine in the measurement of urinary bladder volume. PMID- 15814404 TI - Retroperitoneal solitary fibrous tumour arising from the pelvis in women--a case report and review of literature. AB - A solitary fibrous tumour is an unusual spindle cell neoplasm that most frequently occurs in the pleura based intrathoracic region. In recent years attention has been drawn towards solitary fibrous tumours arising in extrathoracic sites. They are usually benign but malignant solitary fibrous tumours have also been reported (Nielson et al. 1997). There is far less information about the clinical behaviour of an extra thoracic solitary fibrous tumour unlike intrathoracic tumours which is well reported in many case series (England et al. 1989). Although solitary fibrous tumours are well described lesions, the occurrence of similar tumours in the pelvic retroperitoneum of women and presenting as pelvic mass have been reported only sporadically. Because of the rarity, unpredictable behaviour; lack of information available about the clinical behaviour (recurrence and metastasis) and lack of follow up protocol, we are reporting this case which we encountered along with the review of previously reported cases. PMID- 15814405 TI - Tuberous sclerosis in an antenatally diagnosed cardiac rhabdomyoma. PMID- 15814406 TI - Spontaneous maternal adrenal haemorrhage as a cause of intra uterine foetal death. PMID- 15814407 TI - Neonatal necrotising fasciitis and late maternal pelvic abscess formation. A late complication of group A Streptococcus. PMID- 15814408 TI - An unusual case of uterine rupture. PMID- 15814409 TI - Ultrasound-diagnosed placental infarction in a woman with recurrent fetal growth restriction. PMID- 15814410 TI - Anaphylactoid syndrome caused by amniotic fluid embolism following manual removal of placenta. PMID- 15814411 TI - Spontaneous pneumothorax in pregnancy--case report. PMID- 15814412 TI - Chorioangioma: a case report. PMID- 15814413 TI - Heterotopic pregnancy is not rare. A case report and literature review. PMID- 15814414 TI - Secondary abdominal pregnancy associated with uterine fibroid. PMID- 15814415 TI - Tubal abscess in pregnancy: a case report. PMID- 15814416 TI - Severe antepartum haemorrhage following membrane sweep. PMID- 15814417 TI - Difficulties in perinatal diagnosis of fetal thoracic foregut cysts: report of two cases. PMID- 15814418 TI - Postpartum labial adhesions following normal vaginal delivery. PMID- 15814419 TI - Massive vulval oedema complicating pregnancy. PMID- 15814420 TI - Malignancy following treatment of borderline ovarian tumours: latent presentation of a Sister Joseph nodule following treatment of metastatic ovarian adenocarcinoma of the axilla. PMID- 15814421 TI - Adenocarcinoma of the vaginal vault following prolonged unopposed oestrogen hormone replacement therapy. PMID- 15814422 TI - Cervical leiomyosarcoma: a case report. PMID- 15814423 TI - Egg on my face: an unusual way of reducing prolapse. PMID- 15814424 TI - Vesicovaginal fistula caused by a foreign body: delayed presentation and repair with martius graft. PMID- 15814425 TI - A case of diffuse granulomas of the cervix. PMID- 15814426 TI - Bilateral sacrospinous ligament fixation in a patient with Ehlers-Danlos syndrome. PMID- 15814427 TI - Beneficial effects of raloxifene and atorvastatin on serum lipids. PMID- 15814429 TI - Search and destroy: ER quality control and ER-associated protein degradation. AB - Proteins synthesized in the endoplasmic reticulum (ER) encounter quality control checkpoints that verify their fitness to proceed in the secretory pathway. Molecules undergoing folding and assembly are kept out of the exocytic pathway until maturation is complete. Misfolded side products that inevitably form are removed from the mixture of conformers and returned to the cytosol for degradation. How unfolded proteins are recognized and how irreversibly misfolded proteins are sorted to ER-associated degradation pathways was poorly understood. Recent developments from a combination of genetic and biochemical analyses has revealed new insights into these mechanisms. The emerging view shows distinct pathways working in collaboration to filter the diverse range of unfolded proteins from the transport flow and to divert misfolded molecules for destruction. PMID- 15814430 TI - Bacterial small RNA regulators. AB - Small regulatory RNAs can modify the activity of proteins and the stability and translation of mRNAs. They have now been found in a wide range of organisms, and can play previously unsuspected critical regulatory roles. The bacterial small RNAs include two major classes. The largest family(with at least 20 members in Escherichia coli K12) acts by base pairing with target mRNAs to modify mRNA translation or stability; this class of RNAs also uses an RNA chaperone protein, Hfq. DsrA is the best-studied example of this family of RNAs. It has been shown to positively regulate translation of the transcription factor RpoS by opening an inhibitory hairpin in the mRNA, and to negatively regulate translation of hns by pairing just beyond the translation initiation codon. The class of RNAs that modify activity of proteins is exemplified by CsrB and CsrC of E. coli, two RNAs that bind to and inhibit CsrA, a protein translational regulator. Homologs of CsrA and related regulatory RNAs have been implicated in the regulation of gluconeogenesis, biofilm formation,and virulence factor expression in plant and human pathogens. PMID- 15814431 TI - DNA polymerases that propagate the eukaryotic DNA replication fork. AB - Three DNA polymerases are thought to function at the eukaryotic DNA replication fork. Currently, a coherent model has been derived for the composition and activities of the lagging strand machinery. RNA-DNA primers are initiated by DNA polymerase ot-primase. Loading of the proliferating cell nuclear antigen, PCNA, dissociates DNA polymerase ca and recruits DNA polymerase S and the flap endonuclease FEN1 for elongation and in preparation for its requirement during maturation, respectively. Nick translation by the strand displacement action of DNA polymerase 8, coupled with the nuclease action of FEN1, results in processive RNA degradation until a proper DNA nick is reached for closure by DNA ligase I. In the event of excessive strand displacement synthesis, other factors, such as the Dna2 nuclease/helicase, are required to trim excess flaps. Paradoxically, the composition and activity of the much simpler leading strand machinery has not been clearly established. The burden of evidence suggests that DNA polymerase E normally replicates this strand,but under conditions of dysfunction, DNA polymerase 8 may substitute. PMID- 15814432 TI - Platelet-microvessel interactions. PMID- 15814433 TI - Interactions of platelets with subendothelium and endothelium. AB - In this review, the authors summarize how platelets interact with subendothelium when the vessel wall is damaged or with intact endothelium in the inflammatory state. When subendothelium is exposed to rapidly flowing blood upon vessel damage, platelets adhere rapidly to the exposed surface, decelerate, and aggregate to arrest bleeding. Under high shear stress, such as is found in the microcirculation, the interaction between subendothelial von Willebrand factor (VWF) and its platelet receptor, glycoprotein (GP) Ib-IX-V, is required to slow down platelets and allow the platelet collagen receptors aloha2beta1 and GP VI to bind to collagen. GP VI and alpha2beta1 play important roles to activate platelets in the early stage and work with GP Ib-IX-V to fully activate platelets to form thrombi. GP Ib-IX-V and GP VI employ similar signaling pathways for platelet activation and the signals from both receptors are down-modulated by PECAM-1 (platelet-endothelial-cell adhesion molecule 1) to prevent unnecessary platelet activation under high shear. During inflammatory states, intact endothelial cells release VWF and P-selectin from their Weibel-Palade bodies. Both molecules are ligands for GP Ib-IX-V. The newly released VWF is larger and stickier than the form normally found in plasma and binds platelets spontaneously. Normally, VWF is processed by proteolysis by the plasma metalloprotease ADAMTS-13. Failure of this processing results in the microvascular thrombotic disorder thrombotic thrombocytopenic purpura. In this review, the authors also use available crystal structures of platelet receptors and ligands to explain the details of their interactions. PMID- 15814434 TI - Endogenous mechanisms of inhibition of platelet function. AB - Platelets play an important role in coagulation, in maintenance of hemostasis, and in the pathophysiology of thrombotic diseases. In response to blood vessel injury, platelets accumulate at the site, recruit other platelets, promote clotting, and form a hemostatic plug to prevent hemorrhage. By contrast, several inhibitory mechanisms modulate platelet function and act in a synergistic manner to prevent pathologic thrombus formation. This review focuses on the principal endogenous inhibitors of platelet function and the central role of the normal endothelium in these inhibitory processes. The main endothelium-derived platelet inhibitors include nitric oxide, prostacyclin, and Ecto-ADPase/CD39/NTPDase. Each of these factors is discussed in turn, and the specific mechanisms by which they inhibit platelet function are reviewed. PMID- 15814435 TI - Microvascular thrombosis models in venules and arterioles in vivo. AB - Platelets are intimately involved in hemostasis and thrombosis. Under physiological conditions, circulating platelets do not interact with microvascular walls. However, in response to microvascular injury, platelet adhesion and subsequent thrombus formation may be observed in venules and arterioles in vivo. Numerous intravital video microscopy techniques have been described to induce and monitor the formation of microvascular thrombi. The mechanisms of microvascular injury vary widely among different models. Some models induce platelet activation with minimal effects on endothelium, others induce endothelial inflammation or injury, while other models lead to thrombus formation associated with endothelial denudation. The molecular mechanisms mediating platelet-vessel wall adhesive interactions differ among various models. In some instances, differences in responses between venules and arterioles are described that cannot be explained solely by hemodynamic factors. Several models for induction of microvascular thrombosis in vivo are outlined in this review, with a focus on the mechanisms of injury and thrombus formation, as well as on differences in responses between venules and arterioles. Recognizing these characteristics should help investigators select an appropriate model for studying microvascular thrombosis in vivo. PMID- 15814436 TI - Platelet-vessel wall interactions in the microcirculation. AB - Platelet adhesion in the microcirculation is being reported and studied in a growing number of animal models of human disease. The adhesion molecules utilized by platelets to attach to the walls of microscopic blood vessels have been defined in most model systems, with P-selectin-PSGL-1, GPIIb/IIIa-fibrinogen-ICAM 1, and vWF-GPIba interactions serving as major adhesion pathways. The binding of platelets to adherent leukocytes appears to contribute significantly to the platelet-vessel wall interactions elicited in many models of disease. Shear forces generated by the movement of blood through the microcirculation exert an important influence on platelet adhesion. Physiologic regulation of platelet adhesion is mediated by both nitric oxide and superoxide, with the former inhibiting and the latter promoting platelet adhesion. The interactions between platelets and the walls of microscopic blood vessels appear to have important implications in the initiation and/or progression of tissue injury associated with different experimental models of human disease. PMID- 15814437 TI - Regulation of microvascular thromboembolism in vivo. AB - Atherothrombosis and embolization are main causes of morbidity and mortality in the Western world. To optimize treatment, better understanding of the factors involved in thromboembolism in vivo is needed. The course and outcome of a thromboembolic process are determined by the local balance between anti and prothrombotic factors. In healthy vessels, endothelial antithrombotic properties prevent blood platelets from interacting with the vessel wall. Upon vessel wall damage or endothelial activation, however, prothrombotic factors temporarily overrule the antithrombotic factors, leading to thrombus formation and embolization. According to this concept, thromboembolism ends when the balance is restored. Animal models on microvascular thromboembolism have provided evidence that the endothelium is eminently involved in the regulation of thromboembolism, and that shear forces are an important determinant of endothelial function. Therefore, in this review focus is on the endothelial regulation of platelet vessel wall interactions during thromboembolism in vivo. Anti- and prothrombotic properties of vascular endothelium will be discussed, paying special attention to the endothelium-derived platelet inhibiting substances nitiric oxide (NO) and prostacyclin (PGl(2)) and to differences between arteriolar and venular endothelium. In addition, the involvement of shear forces in microvascular thromboembolic processes in vivo will be described PMID- 15814438 TI - Interactions of platelets, blood-borne tissue factor, and fibrin during arteriolar thrombus formation in vivo. AB - Thrombus formation following vascular injury is an essential component of both hemostasis and pathologic vessel occlusion. This process occurs in a closed, pressurized environment in which blood flows rapidly over the injury site. Thrombus formation must occur quickly to reduce blood loss, but is carefully modulated to limit vessel occlusion. Circulating cells, plasma proteins, vessel wall components, and physical forces such as shear all influence thrombus formation. Historically, thrombus formation has been studied by isolating the separate components of blood involved in clot formation. With improved optical techniques, investigators have increasingly studied thrombus formation under conditions of flow in vitro and in live animals in vivo. Using multichannel fluorescence intravital videomicroscopy, the authors have studied the changes in the kinetics and deposition of platelets, fibrin, and tissue factor at the injury site during thrombosis in transgenic mice, bone marrow transplanted mice, and mice treated with pharmacological agents that modulate thrombosis. The differences in the kinetics of accumulation of the various components of thrombus in these mice have provided new insights about thrombus formation in arterioles. This review discusses the role of platelet intracellular signaling, P-selectin expression on platelets, and tissue factor-bearing microparticles in thrombus formation. PMID- 15814439 TI - The impact of neuropsychological impairment and depression on health-related quality of life in HIV-infection. AB - Neuropsychological impairment and depression are common among individual with HIV infection, resulting in significantly altered everyday functioning. The objective of this study was to examine the impact of these two important neurobehavioural complications on health-related quality of life in adults with HIV-infection. Participants (n = 155) received a 3-hours comprehensive neuropsychological examination, the Beck Depression Inventory, and the Medical Outcomes Study HIV Quality of Life instrument. Four groups were formed based on the presence or absence of depression and neuropsychological impairment. Results suggest that neuropsychological impairment and depression can differentially affect dimensions of health-related quality of Life. Specifically, depression has a significant impact on mental health dimensions of health-related quality of life. Some evidence exists for an impact of neuropsychological impairment, or a combined impact of depression and neuropsychological impairment, on the Physical Health dimensions of health-related quality of life. These results confirm the importance of depression as a determinant of health-related quality of life in HIV/AIDS and provide a potential avenue for improving health-related quality of life in adults with HIV-infection. PMID- 15814440 TI - Construct and predictive validity of the repeatable battery for the assessment of neuropsychological status in the evaluation of stroke patients. AB - To evaluate the construct and predictive validity of a brief neuropsychological assessment battery, consecutive admissions to a hospital receiving inpatient treatment for stroke were tested with the Repeatable Battery for Assessment of Neuropsychological Status (RBANS) and other neuropsychological measures. Discriminant and convergent validity were adequate for the Language, Visuospatial/Constructional, Immediate Memory and Delayed Memory indices in the RBANS, but weaker for the Attention Index. Twelve months later, 36 discharged patients were contacted by telephone and interviews using a battery of functional outcome measures. RBANS Total Score predicted indices from the RBANS also predicted cognitive disability as measured by the Functional Independence Measure. Individual indices from the RBANS also predicted instrumental activities of daily living (IADLS) as measured by the Craig Handicap Assessment and Reporting Technique. The present findings suggest that the RBANS can be a useful tool in the neuropsychological evaluation of inpatients with stroke. PMID- 15814441 TI - Cerebral activation patterns during working memory performance in multiple sclerosis using FMRI. AB - Working memory deficits are common in Multi Sclerosis (MS) and have been identified behaviourally in numerous studies. Despite recent advance in functional magnetic resonance imaging (fMRI), few published studies have examined cerebral activations associated with working memory dysfunction in MS. The present study examines brain activation patterns during performance of a working memory task in individual with clinically definite MS, compared to healthy controls (HC). fMRI was performed using a 1.5 Tesla GE scanner during a modified Paced Auditory Serial Addition Test (mPA-SAT). Participants were 6 individuals with MS with working memory impairment as evidenced on neuropsychological testing, 5 individuals with MS without working memory impairment, and 5 HC. Groups were demographically equivalent. Data were analyzed using Statistical Parametric Mapping (SPM99) software, with a stringent significance level (alpha < .005, voxel extent > or =8). Both MS groups and the HC group were able to perform the task, with comparable performance in terms of numbers of correct responses. Activation patterns within the HC and MS not-impaired groups were noted in similar brain regions, consistent with published observations in healthy samples That is, activations were lateralized to the left hemisphere, involving predominantly frontal regions. In contrast, the MS impaired group showed greater right frontal and right parietal lobe activation, when compared with the HC group. Thus, it appears that working memory dysfunction in MS is associated with altered patterns of cerebral activation that are related to the presence of cognitive impairement, and not solely a function of MS. PMID- 15814442 TI - Rey Complex Figure Test performance after traumatic brain injury. AB - The influence of factors such as a cerebral injury characteristics, education, perceptual organization skills, and speed of information processing on performance on the Rey Complex Figure Test & Recognition Trial (RCF) was examined by means of hierarchical regression analyses in 100 consecutively referred persons with traumatic brain injury at a median of 2.5 months post-injury. Patients with premorbid (e.g., psychiatric history) or comorbid (e.g. financial compensation seeking) complicating factors were excluded. Perceptual organization skills and the presence of a diffuse intracranial lesion but not education or speed of processing were statistically significant predictors of the variance in RCF variables. A large proportion of the sample improved by at least a standard deviation from independent delayed recall to multiple-choice recognition, and this was mediated by perceptual organization skills but not by injury parameters. It is concluded that performance on the RCF after traumatic brain injury is affected relatively more by perceptual organization skills than by injury severity characteristics. PMID- 15814443 TI - Cognitive impairment in primary and secondary progressive multiple sclerosis. AB - The aim of this study was to use neuropsychological data to characterize two subtypes of multiple sclerosis (MS) patients in a large patient sample. We studied patients with primary-progressive MS (PPMS) and secondary-progressive MS (SPMS). A group of 121 MS patients (36 PPS, 85 SPMS) and 40 healthy controls were administered a brief battery of cognitive tests. Executive functioning, memory and attention were studied. Results demonstrate that PPMS patients exhibited slightly more impairment than patients with SPMS, although this difference is not significant (50% vs 37%). However, PPMS patients revealed a significantly poorer performance in verbal learning (p < 0.05) and in verbal fluency (p < 0.05). Whereas PPMS patients had significantly shorter disease durations (p < 0.05), there was no statistical difference in disability between both groups. We conclude from our study that cognitive deficits in progressive MS are frequent. Patients with PPMS tend to be more frequently and severely affected than SPMS patients. Our findings of high prevalence of cognitive involvement in PPMS have not been reported previously PMID- 15814444 TI - A meta-analytic review of verbal fluency deficits in depression. AB - A prominent view in the neuropsychological literature is that depression is particularly associated with deficits in executive control processes. A meta analysis of 42 studies with 2306 participants was therefore conducted to investigate the sensitivity of tests of verbal fluency to the pressure of this disorder, as there is a great deal of evidence that theses measures are valid markers of executive dysfunction. When the methodology adopted by other meta analytic reviews was employed, semantic fluency deficits were found to be substantially larger than the phonemic fluency deficits. However, when a more rigorous method of meta-analysis was adopted, this indicated that the measure are in fact broadly equivalent in their sensitivity to depression, as has been found for patients with focal frontal lobe lesions. However, in contrast to patients with focal frontal lobe injuries, neither deficit qualified as a differential deficit relative to psychomotor speed. Therefore, for patients with depression, deficits on tests of phonemic and semantic fluency may not reflect executive dysfunction, but a more generalized impairment. Evidence is presented that tests of phonemic and semantic fluency may aid in the differential diagnosis of patients with depression and those in the early stages of dementia of the Alzheimer's type. PMID- 15814445 TI - A test for measuring recognition and naming of landmarks. AB - There have been considerable giants in recent years in understanding cognitive and neural correlated for retrieval of various types of knowledge, such as the meanings and lexical forms for categories such as a familiar faces, animals and tools, and actions. An important category that has been largely neglected so far is landmarks, and one likely reason for this is a lack of suitable stimulus materials. Here, we report a study in which we designed a Landmark Recognition and Naming Test. The test contains 65 natural (e.g., "Old Faithful") and artifactual (e.g., "U.S Capitol") landmarks from around the world. Preliminary recognition and naming data about the participants (e.g., educational level, participants, experience traveling to national parks), the stimuli (e.g., visual complexity, image agreement), and participant-stimulus interactions (e.g., familiarity, age of acquisition), and analyzed how such factors influenced landmark recognition and naming. There was a pronounced sex-related difference in landmark recognition favoring men; however, men and women performed similarly in landmark naming. We provide here extensive data regarding the stimuli, and we encourage other investigators to make use of our stimuli and data in future investigations of landmark recognition and naming. PMID- 15814446 TI - Corsi Block-Tapping task performance as a function of path configuration. AB - The Corsi Block-Tapping (CB) task has been used as a measure of spatial memory since its development in 1971. However, a standard set of items has been developed for this task, and inconsistencies in performances within levels have been demonstrated in association with different path configuration. This study investigated item consistency by analyzing the performances of 94 young adult participants on a block-tapping task that involved five quasi-randomly determined sequences at each of nice levels of difficulty. In general, performance declined with increasing path length. Cochran Q-test comparisons were conducted on the items within each level, and differential performance were identified at levels 7 and 8 only. Pairwise comparisons determined the specific items for which performance was discrepant, and further analysis indicated that performance decrement were related to more complicated block-tapping paths. The findings suggest that this version of the CB task is relatively consistent overall, and the observed effect of path configuration indirectly corroborated the spatial nature of this task. Performance heterogeneity at higher levels reflected more complicated path configuration and, presumably, greater span capacity load. Differential intra-level item consistency should be considered in clinical applications of spatial (configural) memory tasks in order to avoid erroneous interpretations concerning sustained attention ability based on failures within levels. PMID- 15814449 TI - British Maternal and Fetal Medicine Society 10th Annual Conference. Nottingham, United Kingdom, 14-15 April 2005. Abstracts. PMID- 15814455 TI - Transcarboxylase: one of nature's early nanomachines. AB - The enzyme transcarboxylase (TC) catalyzes an unusual reaction; TC transfers a carboxylate group from methylmalonyl-CoA to pyruvate to form oxaloacetate and propionyl-CoA. Remarkably, to perform this task in Propionii bacteria Nature has created a large assembly made up of 30 polypeptides that totals 1.2 million daltons. In this nanomachine the catalytic machinery is repeated 6-12 times over using ordered arrays of replicated subunits. The latter are sites of the half reactions. On the so-called 12S subunit a biotin cofactor accepts carboxylate, - CO2- , from methylmalonyl-CoA. The carboxylated-biotin then translocates to a second subunit, the 5S, to deliver the carboxylate to pyruvate. We have not yet characterized the intact nanomachine, however, using a battery of biophysical techniques, we have been able to derive novel,and sometimes unexpected, structural and mechanistic insights into the 12S and 5S subunits. Similar insights have been obtained for the small 1.3S subunit that acts as the biotin carrier linking the 12S and 5S forms. Interestingly, some of these insights gained for the 12S and 5S subunits carry over to related mammalian enzymes such as human propionyl-CoA carboxylase and human pyruvate carboxylase, respectively, to provide a rationale for their malfunction in disease-related mutations. PMID- 15814456 TI - mRNA decay in prokaryotes and eukaryotes: different approaches to a similar problem. AB - Over the past 15 years considerable progress has been made in understanding the molecular mechanisms of mRNA decay in both prokaryotes and eukaryotes. Interestingly, unlike other important biological reactions such as DNA replication and repair, many features of mRNA decay differ between prokaryotes or eukaryotes. Even when a particular enzyme like poly(A) polymerase has been conserved, polyadenylation of mRNAs in prokaryotes appears to serve a very different function than it does in eukaryotes. Furthermore, while mRNA degrading multiprotein complexes have been identified in both prokaryotes and eukaryotes, their composition and biochemical mechanisms are significantly different. Accordingly, this review seeks to provide a concise comparison of our current knowledge regarding the pathways of mRNA decay in two model organisms, the prokaryote Escherichia coli and the eukaryote Saccharomyces cerevisiae. PMID- 15814457 TI - Insulin regulation of fatty acid synthase gene transcription: roles of USF and SREBP-1c. AB - The transcriptional regulation of lipogenesis is a highly coordinated process occurring in concert with transcriptional as well as post-transcriptional regulation of enzymes involved in glycolysis and gluconeogenesis. Fatty acid synthase (FAS) plays a central role in de novo lipogenesis by converting acetyl CoA and malonyl-CoA into the final end product, palmitate, which can subsequently be esterified into triacylglycerols and then stored in adipose tissue. Ultimately, this helps to prevent buildup of excess glucose in other types of cells and tissues, the effects of which can be readily observed in the pathophysiology of disease states such as Type-11 diabetes and obesity. Thus, elucidating the transcriptional mechanisms of lipogenic enzyme genes is important for understanding the normal regulation of lipogenesis and ultimately the dysregulation that may occur in certain metabolic disease. In this review, we discuss advances in our understanding of the regulation of lipogenesis at the genetic level, with a special emphasis on the common cis- and trans-acting factors involved in regulation of FAS. Two transcription factors, Upstream Stimulatory Factor (USF) and Sterol Regulatory Element Binding Protein-lc (SREBP lc), seem to play a dominant and possibly cooperative role in regulating FAS transcription. PMID- 15814458 TI - Porphyromonas gingivalis lipopolysaccharide-induced up-regulation in endothelin-1 interferes with salivary mucin synthesis via epidermal growth factor receptor transactivation. AB - Endothelin-I (ET-1) is a 21 amino acid peptide produced from a biologically inactive big ET-1 by the action of endothelin-converting enzyme-1 (ECE-1) that acts through G protein-coupled ETA and ETB receptors. Using mucous cells of sublingual salivary gland, we show that P. gingivalis lipopolysaccharide (LPS) inhibitory effect on salivary mucin synthesis is accompanied by a marked increase in ET-I generation and the enhancement in ECE-1 activity. Inhibition of ECE-I with phosphoramidon led to the impedance of the LPS-induced ET-1 generation as well as countered the detrimental effect of the LPS on mucin synthesis. Moreover, the LPS inhibitory effect of on mucin synthesis was blocked by ETA receptor antagonist, BQ610, but not by ETB receptor antagonist, BQ788. The LPS-induced reduction in mucin synthesis, furthermore, was countered by PD153035 (76.8%), a specific inhibitor of EGFR kinase as well as PP2 (54.7%), a selective inhibitor of tyrosine kinase Src responsible for ligand-independent EGFR transactivation. Our findings are the first to demonstrate that P. gingivalis LPS detrimental effect on salivary mucin synthesis is intimately linked to the events controlled by EGFR transactivation, triggered by upregulation in ECE-1,enhancement in ET-1 production, and G protein-coupled ETA receptor activation. PMID- 15814459 TI - Binding of anti-HIV drugs to human serum albumin. AB - Human serum albumin (HSA), the most prominent protein in plasma, is best known for its exceptional ligand (i.e., drug) binding capacity. Here, values of the dissociation equilibrium constant (Kd)for the binding of HIV protease and reverse transcriptase inhibitors to HSA are reported. The binding of abacavir, atazanavir,didanosine, efavirenz, emtricitabine, lamivudine, nelfinavir,nevirapine, ritonavir, saquinavir, stavudine, zalcitabine, and zidovudine to the Sudlow site I (i.e., the warfarin cleft) located in the subdomain IIA involves the alteration of the HSA structure around Trp214 and induces intrinsic tryptophan fluorescence quenching. Accordingly, ibuprofen that primarily binds to the Sudlow site II located in the subdomain IIIA does not affect the HSA intrinsic tryptophan fluorescence and the binding of anti-HIV drugs to the Sudlow site 1. Accounting for the physiological concentration of HSA (= 7.0 x 10(-4) M), the average anti-HIV drug concentration in plasma (= 1.0 x 10(-4) M), and Kd values for the binding of anti-HIV drugs to HSA (ranging between 4.4 x 10(-5)M and 3.8 x 10(-4) M), it appears that the fraction of HIV protease and reverse transcriptase inhibitors bound to HSA ranges between 63% and 91%. This represents a significant drawback in the anti-HIV therapy and management, the anti-HIV drug concentration required to achieve 90% protease and reverse transcriptase inhibition in the presence of plasma proteins appears to be at least one order of magnitude higher than that required in their absence. PMID- 15814460 TI - In silico search for single nucleotide polymorphisms in genes important in vitamin E homeostasis. AB - Large inter-individual variation exists in the response to vitamin E supplementation, and this may influence the outcome of human studies. It is our hypothesis that genetic heterogeneity is an important determinant of vitamin E homeostasis. Therefore we have performed an in silico search for single nucleotide polymorphisms (SNPs) associated with genes involved in vitamin E homeostasis. Based on function, the following genes were considered as candidates for vitamin E heterogeneity: c-tocopherol transfer protein (TTPA), tocopherol associated protein (TAP), lipoprotein lipase (LPL), multidrug resistance protein 2 (MDR-2), pregnane X receptor (PXR) and members of the cytochrome P450 family (CYP). Searches for coding SNPs were initiated from web based programs of the National Center for Biotechnology Information (NCBI). SNP frequencies were calculated by dividing the number of annotated coding SNPs by the number of base pairs in the open reading frame. Genes for TTPA, TAP and CYP3A5 had calculated SNP frequencies between 503 and 837 base pairs per coding SNP (bp/cSNP) and so are not highly polymorphic. In contrast, cSNP frequencies in LPL, MRP2, PXR, CYP3A4 and CYP4F2 were in the range of 100 bp/cSNP and so are highly polymorphic. Thus proteins involved in specific vitamin E binding are not highly polymorphic, may not influence inter-individual variation and so may not be good candidates for population studies. Proteins involved in drug/lipid metabolism which indirectly influence vitamin E status are highly polymorphic, are likely to influence inter-individual variation and so are good candidates for population studies. We suggest that future studies are aimed at addressing the role of such SNPs in vitamin E homeostasis. PMID- 15814461 TI - Evidence for Na+/Ca2+ exchanger 1 association with caveolin-1 and -2 in C6 glioma cells. AB - The purpose of this study is to understand the interaction of Na + -Ca2+ exchanger (NCX1), that is one of the essential regulators of Ca2+ homeostasis, with caveolin (Cav)-1 and Cav-2 in Cav-3 null cell (rat C6 glioma cell). Both mRNA and protein expression of NCX1, Cav-1 and Cav-2 was observed, but no expression of mRNA and protein of Cav-3 were observed in C6 glioma cells. In isolated caveolae-enriched membrane fraction, the NCX1, Cav-1 and Cav-2 proteins localized in same fractions. The experiment of immuno-precipitation showed complex formation between the NCX1 and Cavs. Confocal microscopy also supported co-localization of NCX1and Cavs at the plasma membrane. Functionally, sodium-free induced forward mode of NCX1 attenuated by Cav-1 antisense ODN. When treated cells with Cav-2 antisense ODN, both reverse and forward mode of NCX1 was attenuated. From these results, in the Cav-3 lacking cells, the function of NCX1 might be regulated by binding with Cavs. Considering the decrement of NCX1 activity by antisense ODNs, caveolins may play an important role in diverse of pathophysiological process of NCX1-related disorders in the body. PMID- 15814463 TI - Is there an answer? PMID- 15814462 TI - HIV protease inhibitors-induced atherosclerosis: prevention by alpha-tocopherol. AB - Prolonged treatments with inhibitors of human immunodeficiency(HIV)-encoded protease (ARPI) have been reported to induce early atherosclerotic events. Our in vitro study indicates that alpha-tocopherol may prevent drug-induced premature atherosclerosis since it interferes with CD36 scavenger receptor over-expression induced by ritonavir in monocytes. The mechanism of CD36 upregulation by ritonavir involves inhibition of the ubiquitin-proteasome system and alpha tocopherol is able to normalize proteasome activity. These findings suggest that ARPI combined with early alpha-tocopherol supplementation may decrease the drug induced atherosclerotic risk. PMID- 15814464 TI - How I became a biochemist. PMID- 15814465 TI - The wars of the carbohydrates: part 3: maltose. PMID- 15814466 TI - Amelioration of endothelin-1-induced optic nerve head ischemia by topical bunazosin. AB - PURPOSE: To investigate the effects of bunazosin hydrochloride, an alpha1 adrenergic blocker, on the impairment of optic nerve head (ONH) blood flow and depression of visual function induced by repeated intravitreal injections of endothelin-1 (ET-1) in rabbits. METHOD: We injected ET-1 (20 pmol) into the right posterior vitreous of rabbits twice a week for 4 weeks, and the observation period was set at 8 weeks (starting the first injection). The animals that received ET01 were divided into two groups: twice a day for 8 weeks, o ne group received topical 0.01% bunazosin, while the second received the vehicle for bunazosin. The ONH blood flow was monitored using the laser speckle method, and visual function was assessed by examining visually evoked potentials (VEPs). Changes in the ONH cup/disk area and in the number of cells in the retinal ganglion cell layer (CCL) were also determined. RESULTS: Repeated injections of ET-1 decreased the ONH blood flow, prolonged the VEP implicit time, enlarged the optic cup, and decreased the number of GCL cells. Topical bunazosin significantly decreased these impairments. CONCLUSIONS: These results indicate that in rabbits, topical bunazosin suppresses the changes in ONH circulation and function induced by intravitreal ET-1. PMID- 15814467 TI - Effects of intravitreal injection of tacrolimus (FK506) in experimental uveitis. AB - PURPOSE: To examine the immunosuppressive and neuroprotective effects of intravitreal injection of tacrolimus in experimental uveitis. METHODS: Tacrolimus (40 microg) was injected intravitreally in rabbits to examine safety. Experimental uveitis was induced in rabbits by systemic immunization with bovine serum albumin (BSA) followed by intravitreal challenge with BSA. On day 1 after BSA challenge, tacrolimus (20 or 40 microg) or betamethasone (400 microg) was injected intravitreally in one eye and balanced salt solution in the contralateral eye. The eyes were evaluated by slit-lamp biomicroscopy, electroretinography, and histopathology. RESULTS: No local or systemic adverse reaction was observed in normal rabbits. In experimental uveitis, intravitreal injection of tacrolimus significantly reduced intraocular inflammation in histopathological analysis (p < 0.03). Amplitudes on the electroretinogram were restored (p < 0.01), and retinal thickness was preserved in tacrolimus-treated eyes (p < 0.03). CONCLUSIONS: In experimental uveitis, intravitreal injection of tacrolimus effectively suppresses ocular inflammation and preserves retinal architecture. PMID- 15814468 TI - Carboxyamido-triazole modulates retinal pigment epithelial and choroidal endothelial cell attachment, migration, proliferation, and MMP-2 secretion of choroidal endothelial cells. AB - PURPOSE: To determine the effect of the calcium signaling modulating drug carboxyamido-triazole (CAI) on substeps of exudative age-related macular degeneration (AMD) in vitro. MATERIALS AND METHODS: Zymography and ELISA determined the effect of CAI on MMP-2 production of choroidal endothelial cells (CECs) stimulated by bFGF and VEGF. The effects of CAI on attachment of retinal pigment endothelial (RPE) cells/CECs onto fibronectin, laminin, collagen IV, and migration toward fibronectin were investigated. Proliferation induced by serum and bFGF (10 microg/ml) with and without CAI (0.1-10 microM) was measured by cell counting and 3H-uptake. Viability and apoptosis of the exposed cells was assessed by an MTT and an apoptosis assay. RESULTS: CAI inhibited serum- and bFGF-induced proliferation, cell attachment onto fibronectin and collagen IV, but only CEC attachment onto laminin. Inhibition of MMP-2 production was observed (10 microM CAI). CAI reduced the cellular viability by apoptosis induction. CONCLUSIONS: CAI inhibits substeps of exudative macular degeneration and may be of value for the treatment of the disease. PMID- 15814469 TI - Interleukin-4-mediated infiltration of eosinophils into the conjunctiva and its suppression by interferon-gamma. AB - PURPOSE: Interleukin (IL)-4 is a T helper (Th)2 cytokine that plays an important role in the development of allergic reactions. It has been suggested that IL-4 is responsible for the infiltration of eosinophils into the conjunctiva during the development of allergic conjunctivitis. However, it is still unclear whether IL-4 is able to induce this eosinophilic infiltration on its own. We investigated whether subconjunctival injection of IL-4 can induce eosinophils to infiltrate into the conjunctiva. METHODS: Brown Norway rats were subconjunctivally injected with IL-4, eotaxin, or phosphate buffered saline (PBS), and the conjunctivas were harvested for histologic analysis (including immunohistochemistry) 6, 12, 18, and 24 hr later. The harvested conjunctivas were also subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to examine chemokine expression. In a separate experiment, the effect of coadministering interferon gamma (IFN-gamma) along with IL-4 was examined. RESULTS: The subconjunctival injection of IL-4 induced eosinophil infiltration into the conjunctiva in a dose dependent manner. IL-4 was as potent as eotaxin. The eosinophilic infiltration started 6 hr after the injection and persisted for up to 24 hr after the injection. Other infiltrating cell phenotypes were noted but were also observed in conjunctivas injected with PBS alone. RT-PCR analysis demonstrated that IL-4 injection did not upregulate chemokine RNA expression in the conjunctiva. Coinjection of IFN-gamma suppressed the infiltration of eosinophils into the conjunctiva induced by IL-4 injection. CONCLUSIONS: The subconjunctival injection of IL-4 specifically induces eosinophils to infiltrate into the conjunctiva. In addition, IFN-gamma in the conjunctiva can counteract this effect of IL-4. PMID- 15814470 TI - Effect of cataract surgery with intraocular lens implant on frequency doubling perimetry. AB - PURPOSE: To study the effect of cataract surgery with intraocular lens (IOL) on frequency doubling perimetry (FDP). METHODS: Patients aged 40 years or above seen at our outpatient clinic with no ocular pathology except for visually significant cataract and visual acuity 6/24 or better were eligible. They underwent FDP before and 4 to 6 weeks after cataract surgery with IOL. RESULTS: Screening test: Mean scores by three different scoring methods were 1.82 (3.21), 2.80 (5.54), 4.18 (9.18) before and 0.22 (0.51), 0.26 (0.63), 0.26 (0.69) after surgery (p = 0.002 0.001, < 0.0001). Threshold test: Mean deviation (MD) and pattern standard deviation (PSD) were -5.23 (3.08) and 5.15 (2.78) before and -2.94 [corrected] (2.49) (p < 0.0001) and 5.21 (1.780) (p = 0.63) after surgery. CONCLUSIONS: The screening test should be interpreted cautiously in the presence of cataract. On threshold testing, cataract surgery causes significant decrease in MD but no change in PSD. PMID- 15814471 TI - Dorzolamide influences the autoregulation of major retinal vessels caused by artificial intraocular pressure elevation in patients with POAG: a clinical study. AB - PURPOSE: The study investigated whether dorzolamide influences the autoregulatory behavior of major retinal arterioles in glaucoma patients via a moderate perfusion pressure reduction. METHODS: The study included one eye each of 12 untreated patients with a primary open-angle glaucoma (POAG) (age 60.8 +/- 8.3, IOP 22.3 +/- 6.5 mmHg). Changes in the diameter of a retinal artery segment before (120 s), during (100 s), and after (380 s) artificial IOP elevation to 38 mmHg for 100 s were recorded continuously by means of a Retinal Vessel Analyzer. The measurement was repeated after 4-week treatment with dorzolamide eye drops t.i.d. RESULTS: Ocular perfusion pressure (mmHg) was reduced by the intraocular pressure (IOP) elevation from 58 (+/- 10) to 41 (+/- 11) in the pretreatment examination and from 60 (+/- 8) to 40 (+/- 8) posttreatment (differences between the examinations n.s.). Before IOP elevation, the arterial diameter was found to be +1.7 +/- 3.5% greater in the posttreated eyes than in the pretreated eyes (p < 0.02). During IOP elevation, the arterial diameter decreased by -1.8% +/- 3.8 in the pretreated eyes, whereas dilatation by +1.4% +/- 2.5 was observed in the posttreated eyes (p = 0.02). At the end of the observation period following IOP elevation, the vessel diameter in the pretreated eyes had increased by +1.8% +/- 4.2, whereas in the posttreated eyes it had decreased by -1.7% +/- 3.0. On average, dorzolamide reduced IOP by -5.6 mmHg (p = 0.001). CONCLUSIONS: The arterial diameter dilatation during IOP elevation in dorzolamide-treated eyes could be an accelerated counter-regulation on the induced elevated IOP and could constitute an additional therapeutic effect. PMID- 15814472 TI - Clozapine directly relaxes bovine retinal arteries. AB - PURPOSE: It was suggested that clozapine might be helpful in the development of new antiglaucoma agents, as it combines lowering the intraocular pressure after topical instillation with vasodilation. This study aimed to evaluate and characterize the vasodilatory effect of clozapine in isolated bovine retinal arteries (BRAs). METHODS: Retinal arteries were isolated from bovine eyes and mounted in the organ bath of a small vessel myograph. RESULTS: Cumulative addition of clozapine (1 nM to 10 microM) caused a concentration-dependent relaxation of the BRAs. Removal of the endothelium, inhibition of nitric oxide synthase and of soluble guanylyl cyclase reduced the clozapine response, whereas cyclooxygenase inhibition had no influence. A Ca2+ channel activator, a 5 hydroxytryptamine receptor antagonist, and an adenosine receptor antagonist failed in affecting the clozapine-induced relaxations. CONCLUSIONS: Clozapine relaxes bovine retinal arteries. Endothelium-derived NO seems to be involved, whereas prostanoids, calcium entry blockade, 5-HT7 receptor stimulation, and adenosine receptor stimulation do not. PMID- 15814473 TI - Interaction between surgical procedure for repairing retinal detachment and clinical risk factors for proliferative vitreoretinopathy. AB - PURPOSE: To asses risk factors of proliferative vitreoretinopathy (PVR) and a model for predicting it. METHODS: Observational, case-control. 335 patients with non-complicated retinal detachment (RD) were included: 134 developed PVR (Cases); 201 patients did not (Controls). Risk factors for PVR were identified by multivariate analysis. Influence of variables was assayed according to the surgical approach. By logistic regression analysis a model to predict the risk of developing PVR and odds ratio (OR) values for each clinical factor were estimated. RESULTS: Risk was higher in patients > 70 years and with intraocular pressure lower than 14 (OR: 3.84; CI 95%: 2.04-7.30) and in retinal breaks larger than "1 clock hour" (OR: 2.54; CI: 1.28-5.05), extended retinal detachments (OR: 4.01; CI: 1.98-8.10) and reinterventions (OR: 1.55; CI: 1.14-9.22). Scleral surgery also was a risk factor (OR: 3.89; CI: 2.12-7.14) and aphakia/pseudophakia when scleral surgery is performed (OR: 3.33; CI: 1.54-7.22). A model to predict PVR was proposed with these results. CONCLUSIONS: Surgical approach modifies risk factors of PVR, and should be taken into account to improve the models for predicting it. PMID- 15814475 TI - Behavioral Dyscontrol Scale-Electronic Version: first examination of reliability, validity, and incremental utility. AB - Behavioral Dyscontrol Scale (BDS) is a clinical measure previously shown to be related to frontal lobe integrity, executive abilities, and functional independence. Electronic version of the scale (BDS-EV) was developed and its reliability and validity were examined. The BDS-EV, the original BDS, and a brief battery of traditional clinical tests were administered to 55 community-dwelling adults ages 18 to 68. The results yielded high internal consistency and provided support for convergent, discriminant, and incremental validity. Overall, the results demonstrate the feasibility of converting the BDS into an electronic instrument and support continued research and development of this instrument. PMID- 15814476 TI - Base rates of longitudinal RBANS discrepancies at one- and two-year intervals in community-dwelling older adults. AB - Identification of clinically significant change in performance over time on neurocognitive tests is an important aspect of neuropsychological evaluation; however, scant published empirical data exists to guide the clinician in determining the significance of psychometric change across clinically relevant retest intervals. The present study presents base rate data of RBANS score discrepancies in a user-friendly manner based on the performances of a large sample (n=283) of community-dwelling older adults. Data for 1- and 2-year retest intervals are presented in a tabular form that can be used as a convenient reference. Base rates of discrepancy scores were calculated and organized into three groups (i.e., below average, average, and above average) with respect to the participants' OKLAHOMA age- and education-corrected RBANS Total Scale score (Duff, Patton, Schoenberg, Mold, Scott, & Adams, 2003) at initial assessment, in an effort to reduce the influence of regression to the mean and practice effects that is associated with varying levels of cognitive ability. (e.g., Rapport, Axelrod, Theisen, Brines, Kalechstein, & Ricker, 1997; Rapport, Brines, Axelrod, & Theisen, 1997). These data may be helpful in clinical practice by assisting the clinician in determining the clinical significance of score changes. PMID- 15814477 TI - Demographic influences on baseline and derived scores from the trail making test in healthy older Australian adults. AB - Normative data from 363 community-dwelling older adults (60-89 years) is presented in relation to performance on the Trail Making Test. Age and years of education were identified as significantly impacting on performance. Therefore, data are presented for four separate age groups (60-69; 70-74; 75-79; 80-89) and two different lengths of education. In addition to data for Trails A and B, information is provided for the difference and ration scores, which are increasingly used by clinicians to interpret executive function. PMID- 15814478 TI - Circadian preference and cognitive functioning among rehabilitation inpatients. AB - The influence of circadian preference was examined among 56 morning-oriented rehabilitation inpatients with cognitive (n=28) and noncognitive (n=28) impairments. Each individual was tested twice: morning (preferred time) and evening (nonpreferred time); sessions and test batteries were counterbalanced to control for practice effects. Standard measures assessed attention, language, memory, visuospatial, and executive functions. Persons with cognitive impairment showed disproportionate vulnerability to the effects of circadian preference and time of testing, performing more poorly at nonpreferred than preferred times. Substantial effects (eta2 .12 to .48) were found on tests of executive functioning and tasks incorporating similar higher-order demands (e/g/. complex figure copy). Results are supported by tympanic temperature changes during a vigilance task, and index of cerebral blood flow in response to cognitive challenge. Cognitive reserve theory is suggested as an explanation for the differential effects. These findings may have implications for inpatient therapeutic interventions and discharge planning. PMID- 15814479 TI - Development of the WAIS-III general ability index estimate (GAI-E). AB - The WAIS-III General Ability Index (GAI; Tulsky, Saklofske, Wilkins, & Weiss, 2001) is a recently developed, 6-subtest measure of global intellectual functioning. However, clinical use of the GAI is currently limited by the absence of a method to estimate premorbid functioning as measured by this index. The purpose of this study was to develop regression equations to estimate GAI scores from demographic variables and WAIS-III subtest performance. Participants consisted of those subjects in the WAIS-III standardization sample that has complete demographic data (N=2,401) and were randomly divided into two groups. The first group (n=1,200) was used to develop the formulas (i.e., Development group) and the second (n=1,201) group was used to validate the prediction algorithms (i.e., Validation group). Demographic variables included age, education, ethnicity, gender and region of country. Subtest variables included vocabulary, information, picture completion, and matrix reasoning raw scores. Ten regression algorithms were generated designed to estimate GAI. The GAI-Estimate (GAI-E) algorithms accounted for 58% to 82% of the variance. The standard error of estimate ranged from 6.44 to 9.57. The correlations between actual and estimated GAI ranged from r=.76 to r=.90. These algorithms provided accurate estimates of GAI in the WAIS-III standardization sample. Implications for estimating GAI in patients with known or suspected neurological dysfunction is discussed and future research is proposed. PMID- 15814480 TI - Mini-Mental State and Cognistat performance in an older African American sample. AB - Ninety-three normal, healthy, community-dwelling African American men and women aged 60 and older completed the Folstein Mini-Mental State Examination (MMSE) and the Cognistat (formerly known as Neurobehavioral Cognitive Status Examination). Results indicated significant age and education effects on MMSE and Cognistat performance. The strength of the effects was less than had been noted in earlier work. Cognistat scores were lower than those reported in the original manual for the test as well as in comparison to an older sample in another study. Explanations were offered in terms of differences in cultural and educational exposure to testing as well as inappropriateness of the norms in the original manual. Age- and education-stratified normative data are presented that should be helpful to clinicians who work with and perform cognitive assessment in the population. PMID- 15814481 TI - Agreement between the abbreviated and standard portland digit recognition test. AB - This study sought to determine the classification agreement between the standard and abbreviated forms of the Portland Digit Recognition Test (PDRT), a well established symptom validity test. PDRTs (N=200: 100 traumatic brain injury, 100 chronic pain) were randomly selected from case manager and attorney referrals for psychological assessment. Only three cases were misclassified and agreement was as high as 99.5%. This study demonstrated that when using the abbreviated PDRT there is little risk of false negative errors. PMID- 15814482 TI - Sensitivity and specificity of finger tapping test scores for the detection of suspect effort. AB - Past studies indicate that patients with incentive to fake neuropsychological symptoms are likely to have lower finger tapping scores than credible patients. The present study builds upon past research by investigating finger tapping performance for seven groups: (a) noncredible patients (as determined by failed psychometric and behavioral criteria), and patients with (b) closed head injury, (c) dementia, (d) mental retardation, (e) psychosis, or (f) depression, and (g) healthy older controls. Results showed that men tapped faster than women, requiring that groups be divided by gender. Noncredible male and female patients tapped slower than their comparison group counterparts. Dominant hand score proved to be more sensitive to noncredible performance than other scores (nondominant, sum of both hands, difference between dominant and nondominant), especially for women. Sensitivity, specificity, and positive and negative predictive value tables are presented. With specificity set at 90% for the comparison groups combined, a dominant hand cutoff score of /=3 errors was the best predictor of group status. Malingering research employing a group of probable clinical malingerers has direct generalizability to real-world settings. PMID- 15814487 TI - Mechanisms involved in A/J mouse lung tumorigenesis induced by inhalation of an environmental tobacco smoke surrogate. AB - Lung tumors have been reproducibly induced in A/J mice exposed to a surrogate for experimental environmental tobacco smoke (ETSS) in a 5-mo inhalation period followed by 4 mo without further exposure. In order to increase our mechanistic understanding of this model, male mice were whole-body exposed for 6 h/d, 5 d/wk to ETSS with a particulate matter concentration of 100 mg/m(3). Food restriction regimens were included to model or exceed the ETSS-related impairment of body weight development. Half of the mice were pretreated with a single ip injection of urethane to study the effect of the above treatments on lung tumor development induced by this substance. At 5 mo, the tumor response was statistically the same for all groups of non-pretreated mice; however, the expected urethane-induced lung tumorigenesis was significantly inhibited by approximately 25% by ETSS and food restriction. This inhibition was accompanied by a threefold increase in blood corticosterone as a common stress marker for both ETSS and food restriction. At 9 mo, in mice not pretreated, the lung tumor incidence and multiplicity were significantly increased by twofold in the ETSS group; in the urethane-treated groups, the same high tumor multiplicity was reached regardless of previous treatment. The predominant tumor type in all groups was bronchiolo alveolar adenoma. There was no induction of a specific K-ras mutation pattern by ETSS exposure. These data suggest a stress-induced inhibition of lung tumorigenesis in this model, explaining the need for the posttreatment period. PMID- 15814488 TI - Colchicine decreases airway hyperreactivity after phosgene exposure. AB - Phosgene (COCl(2)) exposure affects an influx of inflammatory cells into the lung, which can be reduced in an animal model by pretreatment with colchicine. Inflammation in the respiratory tract can be associated with an increase in airway hyperreactivity. We tested the hypotheses that (1) phosgene exposure increases airway reactivity and (2) colchicine can decrease this elevation. Sprague Dawley rats (70 d old; male) were exposed to 1 ppm COCl(2) for 1 h. Airway reactivity was tested at 0, 4, and 24 h postexposure by infusing anesthetized animals intravenously with acetylcholine and assessing expiratory resistance and dynamic compliance. Immediately and 4 h postexposure, a significant change in expiratory resistance and dynamic compliance was observed in those animals exposed to COCl(2), while at 24 h this response was greater. A second experiment was performed in rats pretreated with colchicine (1 mg/kg) or saline given intraperitoneally, exposed to 1 ppm COCl(2) for 1 h, with both expiratory resistance and dynamic compliance assessed at 24 h. After exposure, cell differentials and protein in lavage were also quantitated. The results indicate that colchicine decreased neutrophil influx, protein accumulation, and changes in both expiratory resistance and dynamic compliance after COCl(2) exposure. Colchicine may affect injury and changes in expiratory resistance and dynamic compliance by diminishing the incursion of inflammatory cells, but other properties of this medication may also be responsible for the observed results. PMID- 15814489 TI - A quantitative in vitro fluorescence imaging method for phospholipid loss from respirable mineral particles. AB - Respirable quartz and kaolin particles were treated with fluorescent-labeled phospholipids to model contact of fibrogenic and nonfibrogenic particles with pulmonary surfactant in the alveolar regions of the lung. Particles were used to challenge rat pulmonary macrophages in vitro at times from 1 d to 10 d. The objective was to develop a quantitative method to track surfactant components that adsorb to respirable particles in the lung or inside cells. Confocal laser scanning microscopy was used to image and quantify surfactant remaining on particles internalized by cells. Results indicate that the fluorescent label is removed from quartz particles quickly, with the fluorescence intensity less than 15% of initial value at 3 d, and about 5% at 10 d. In contrast, the kaolin particle-associated fluorescence was still approximately 39% of initial intensity at 3 d, and 10-15% at 10 d. Unchallenged cells showed a background of approximately 5%, and noninternalized particles did not exhibit any loss of fluorescence over the 10-d exposure. The results indicate the method may be useful in label-removal rate studies of respirable particles in vitro, with some cautions and limitations. Results are discussed and compared with similar studies using nonimaging techniques. PMID- 15814490 TI - Expression of heme oxygenase-1 in the lungs of rats exposed to crocidolite asbestos. AB - Oxidative stress is thought to be the pathogenesis of pulmonary fibrosis induced by asbestos, and heme oxygenase-1 (HO-1) protects lung tissue against oxidative stress. We hypothesized that HO-1 is associated with oxidative lung injury caused by exposure to asbestos. This study was conducted to investigate the time course of HO-1 expression of lungs exposed to crocidolite asbestos in vivo. Male Wistar rats were administered 1 mg or 2 mg crocidolite asbestos suspended in saline by a single intratracheal instillation and were sacrificed at 3 d, 1 wk, 1 mo, 3 mo, and 6 mo of recovery time. The expression of HO-1 was observed by Western blot analysis and immunostaining. Protein levels of HO-1 increased at from 3 d to 6 mo following intratracheal instillation of 2 mg crocidolite asbestos. The levels of HO-1 increased at 1 wk and 1 mo following intratracheal instillation of 1 mg crocidolite asbestos. Many HO-1-positive cells were found, particularly in the alveolar macrophages, during immunostaining. These findings suggest that HO-1 may be related to lung disorder induced by dust and therefore can act as a biomarker of lung injury due to dust exposure. PMID- 15814491 TI - Neutrophils potentiate platinum-mediated injury to human ciliated epithelium in vitro. AB - Exposure to platinum salts, such as may occur in the platinum refining industry, can be associated with the development of airway disorders such as asthma. However, there have been no studies investigating the direct effects of platinum salts on human ciliated epithelium. We have investigated the effects of platinic chloride on human ciliated epithelium, obtained by brushing the inferior nasal turbinate of healthy human volunteers. Ciliary beat frequency was measured using a phototransistor technique, and damage to the structural integrity of the epithelium was measured using a visual scoring index. Platinic chloride at concentrations between 0.25 and 25 microM caused a dose-dependent slowing of ciliary beating and damage to the structural integrity of the epithelium. These direct injurious effects were not affected by catalase, but were almost completely attenuated by preincubation of the epithelium with cysteine. The effects of platinic chloride on ciliary beating and structural integrity were enhanced by the presence of neutrophils and were partially attenuated by preincubation of the epithelial strips with catalase, suggesting that the direct effects of the metal were enhanced in this experimental system by reactive oxidants produced by activated neutrophils. This study documents that platinum salts have an injurious effect on human ciliated epithelium in vitro. If such effects also occur in vivo they may play a role, at least partly, in the pathogenesis of airway disorders that may manifest in exposed workers. PMID- 15814492 TI - Effect of sulfur dioxide inhalation on cytokine levels in lungs and serum of mice. AB - In order to elucidate the immunotoxic mechanism exerted by sulfur dioxide (SO(2)), we investigated the effect of SO(2), a major air pollutant, on the cytokine levels in lungs and serum of male mice. Levels of interlukin-6(IL-6), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta1 (TGF-beta1) in lungs and serum from male mice exposed to SO(2) at various concentrations were measured by the enzyme-linked immunosorbent assay. Sixty Kunming albino male mice were divided randomly into six equal groups: three groups exposed to SO(2) (14.00 +/- 1.25, 28.00 +/- 1.98, and 56.00 +/- 3.11 mg/m(3), which are 5 +/- 0.45, 10 +/- 0.71, and 20 +/- 1.11 ppm) and their respective control groups. The results were as follows: (1) For lung tissues of male mice, exposure to SO(2) at 14 mg/m(3) (5 ppm) caused statistically significant increase of levels of IL-6 and TNF-alpha (p < .05) compared with the control group; exposure at 28 mg/m(3) (10 ppm) caused a statistically highly significant increase of level of IL-6 (p < .01) and a significant increase of TNF alpha (p < .05); and exposure at 56 mg/m(3) (20 ppm) caused no any significant increase of levels of IL-6 and TNF-alpha. SO(2) at all concentrations tested could not cause significant change of level of TGF-beta1 in lungs. (2) For serum from male mice, after exposure to SO(2) at 14 mg/m(3) (5 ppm), the level of TNF-a was significantly increased (p < .05) compared with the control group, but the changes of levels of IL-6 and TGF-beta1 were not significant. After exposure to SO(2) at 28 mg/m(3) (10 ppm) and 56 mg/m(3) (20 ppm), levels of IL-6 and TNF alpha were increased nonsignificantly, but the level of TGF-beta1 was decreased nonsignificantly. These results imply that inflammation reaction could be induced in lung tissue by SO(2) inhalation and the inflammation reaction might relate to these cytokines. And determination of cytokines in lung may be more valuable than in serum when lung injury caused by SO(2). PMID- 15814493 TI - Levels of sulfite in three organs from mice exposed to sulfur [corrected] dioxide. AB - To study whether sulfur dioxide (SO(2)) can enter into the different organs of mice exposed to SO(2), the sulfite contents in brains, hearts, and lungs from male mice were determined by high-performance liquid chromatography with fluorescence detection (HPLC-FD). After reduction and precolumn derivation of tissue homogenates of brains, hearts, and lungs from mice, the mixture was centrifuged, and 5 microl of the resulting supernatant was directly injected into HPLC; the mobile phase consisted of methanol-phosphoric acid (12:88, v/v), and for the fluorescence detection lambdaEX 392 nm and lambdaEM--479 nm were used. The standard curve was linear in the range from 0.126 microg/ml to 126 microg/ml; the minimal detectable concentration was 0.04 microg/ml (S/N = 3), the average methodological recoveries were from 97% to 101%, and the within-day and between day precisions were less than 9%. These results showed that sulfite contents in all organs tested from mice in the SO(2)-exposed groups were significantly increased (p < .05) in a dose-dependent manner (r > .92) compared with the control groups. These results indicated SO(2) could transform into sulfite in vivo after inhalation, and could distribute into lung and other organs such as brain and heart. These results offered a support for the viewpoint that SO(2) is a systemic toxic agent. PMID- 15814497 TI - International Lesbian and Gay Law Association. PMID- 15814495 TI - Determination of closed form solution for acceptance sampling using ANN. AB - Tabled sampling schemes such as MIL-STD-105D offer limited flexibility to quality control engineers in designing sampling plans to meet specific needs. We describe a closed form solution to determine the AQL indexed single sampling plan using an artificial neural network (ANN). To determine the sample size and the acceptance number, feed-forward neural networks with sigmoid neural function are trained by a back propagation algorithm for normal, tightened, and reduced inspections. From these trained ANNs, the relevant weight and bias values are obtained. The closed form solutions to determine the sampling plans are obtained using these values. Numerical examples are provided for using these closed form solutions to determine sampling plans for normal, tightened, and reduced inspections. The proposed method does not involve table look-ups or complex calculations. Sampling plan can be determined by using this method, for any required acceptable quality level and lot size. Suggestions are provided to duplicate this idea for applying to other standard sampling table schemes. PMID- 15814498 TI - CERSGOSIG: perspectives and objectives to challenge discrimination. A network on global scale. PMID- 15814499 TI - Sexuality and international human rights law. AB - This essay considers the extent to which international human rights now protect, or might protect, GLBT communities. The counterpoint between the potential width of application of international human rights instruments and their silence on sexuality has become the leitmotif of sexuality and gender identity within the international human rights framework. In addition, there is a symbiotic relationship between the international norms and domestic legal systems which directly affects the meaning of those norms. Domestic laws are not only needed to implement international norms, but are essential in overcoming the equivocations and silences of international human rights law as it has traditionally applied to GLBT communities. A fusion of the international norms with domestic legal systems through the principle of diversity, rather than the principle of equality, is needed. PMID- 15814500 TI - Sexuality and Australian law. AB - The author describes the changing legal environment concerning same-sex relationships in the common law world with special reference to Australia. He refers to shifts in public opinion recorded in opinion polls; important decisions of human rights courts and tribunals; and changes in national law and court decisions. He then reviews the Australian constitutional setting which divides lawmaking responsibility on such subjects between the federal, State and Territory legislatures. He describes initiatives adopted in the States and Territories and the more modest changes effected in federal law and practice. He concludes on a note of optimism concerning Australia's future reforms affecting discrimination on the grounds of sexuality. PMID- 15814501 TI - Transsexuals and European human rights law. AB - Sexual identity is a legal status, and as such it is as much dependent on public policy as on self-identification. However, because this status can be crucial to one's role in society, a conflict between the legal status and an individual's perceptions or aspirations creates a dilemma if society is committed to individual freedom. This difficulty can become particularly acute where it is technically possible for an individual to alter some of the factors used socially or legally to determine sexual identity. This paper analyses these difficulties, both from the situation of endocrine disorder and with respect to gender identity. It argues that this distinction in approach may not be a valid basis for different legal treatment. It considers four major issues: the stage at which a change of gender should be recognised; any preconditions to which a legal change of gender should be made subject; whether legal recognition should be made for all purposes or only for specific areas of the law; and the confidentiality of a person's previous sexual identity. The paper particularly analyses the law in the United Kingdom and then considers other jurisdictions before focusing on the treatment of transsexuals under the European Convention on Human Rights. PMID- 15814502 TI - Sexual orientation and gender identity in North America: legal trends, legal contrasts. AB - The article provides a comparative analysis of recent legal developments with regard to LGBT rights in Canada and the United States of America. Both countries have made great progress towards better protection and full recognition of LGBT rights. Despite this progress, Canada appears to be ahead of the United States, as illustrated by two most recent court decisions (e.g. Halpern, legalizing same sex marriage in Ontario, and Lawrence v. Texas, decriminalizing consensual sodomy in the United States). PMID- 15814503 TI - Sexuality and human rights in europe. AB - Written human rights law in Europe is as scanty as in the rest of the world. Case law however provides considerable protection of sexual rights. It guarantees comprehensive protection of autonomy in sexual life, also for minors, and provides protection against discrimination based on sexual orientation. Negative attitudes of a majority may not justify interferences with the sexual rights of a minority and society could be expected to tolerate a certain inconvenience to enable individuals to live in dignity and worth in accordance with the sexual identity chosen by them. Compensation for interference with sexual autonomy and freedom is awarded. This high-level protection (as compared to other parts of the world) is however limited. It seems to be granted only in areas where it corresponds with public attitudes and social developments. And it is seldom secured on the national level but nearly exclusively by the European Court of Human Rights, whose case-law is often weakened by inconsistency. PMID- 15814504 TI - Advancing human rights through constitutional protection for gays and lesbians in South Africa. AB - As a consequence of the 1994 adoption of a justiciable Bill of Rights in South Africa, with an equality provision prohibiting discrimination on the ground of sexual orientation, a coalition of gay and lesbian organisations set about implementing a progressive agenda of gay and lesbian rights litigation. In striking down the offence of sodomy, the Constitutional Court established a jurisprudence of gay and lesbian rights to equality, dignity and privacy that proved to be the foundation for significant litigation around family law issues. Subsequent to the sodomy judgement, the Court has ruled that same-sex couples who are in permanent life partnerships should be entitled to the same rights as married couples to immigration, employment benefits, custody and adoption of children. Despite the extensive equality jurisprudence of the Court, it is still uncertain whether it will rule in the future in favour of same-sex marriage or in favour of a civil union/domestic partnership model. PMID- 15814505 TI - Sexuality and human rights: an Asian perspective. AB - In Asia, the lesbian and gay rights movements are clearly dominated by activists, who tend to think in terms of a binary opposition (homo- vs hetero-) and clear cut categories. Based on "Western patterns," the approach is practical, the arguments based on minority rights. "Coming out" is often perceived as a "white model" bringing more problems than real freedom. On the contrary, "Asian values" put the emphasis on family and social harmony, often in contradiction to what is pictured as "lesbian and gay rights." Homophobia follows very subtle ways in Asian countries. Asian gays have to negotiate their freedom, lifestyle and identities in an atmosphere of heterosexism, and not the endemic violent homophobia prevalent in many western countries. In Asia, one's identity relates to one's position in the group and sexuality plays a relatively insignificant role in its cultural construction. That Asian gays often marry and have children shows the elasticity their sexual identity encompasses. Fluidity of sexuality does not really match the Western approach in terms of essentialist categories that have a right to exist. Most Asian societies can be thought of as "tolerant" as long as homosexuality remains invisible. Procreative sexuality can be seen as a social duty, and heterosexual marriage is often not considered incompatible with a "homosexual life." The development of the Internet has even facilitated the encounters while allowing secrecy. Unfortunately, the traditional figures of transgender and transvestites have often been separated from the gay liberation movement. PMID- 15814506 TI - Laws and sexual identities: closing or opening the circle? AB - This essay considers, in comparative perspective, the articles in this volume to see what lessons might be learned with respect to the effectiveness of human rights norms in the struggle for GLBT rights and how these lessons might be used for successful activism. It notes that the concept of "GLBT" is itself not uniform, so that "GLBT rights" must also be contextual. It argues that successful change has been wrought when GLBT rights, however perceived, have been built onto the earlier successes of other movements, such as those for racial and gender equality. This in turn requires, and is assisted by, throwing off parochialism which prevents or hampers the building on the successes that may have occurred in other jurisdictions. Activism needs to be fitted to context, so that a "critical mass" is achieved, allowing sufficient acceptance of change. Once this is translated into law, a "halo" effect is produced. However, this is not the end of the problem, but the beginning of a new set of problems. PMID- 15814508 TI - Pain medicine and drug law enforcement: an important step toward balance. PMID- 15814509 TI - Does the DEA truly seek balance in pain medicine? A chronology of confusion that impedes good patient care. PMID- 15814510 TI - Journal innovations: cover art to reflect suffering and relief from pain and related symptoms--a patient consultation feature, and an annual award for the best paper in the journal. PMID- 15814511 TI - Progress to achieve balanced state policy relevant to pain management and palliative care: 2000-2003. AB - State laws and regulatory policies govern healthcare practice, including the prescribing, dispensing, and administering of opioid analgesics to treat pain. A number of national healthcare and law enforcement organizations have identified drug regulatory policy as a potential barrier to pain relief and palliative care, and have called for evaluation and removal. This article summarizes and discusses the results of an innovative evaluation methodology that was used to produce three policy analysis tools, including one report that graded and ranked states based on the quality of their policies related to pain management and palliative care (called a Progress Report Card [PRC]). The PRC development and implementation was a first-of-a-kind study that compared pain policies in all states over a three year period according to the same evaluation criteria. Results demonstrate significant progress to improve policy in a number of states during the study period, but also showed that most state policies are characterized by a lack "balance." In addition to providing examples of policy change in particular states, the relevance of these findings to current policy issues, including the importance of communicating and implementing new policies is discussed. The need for partnerships between the healthcare and law enforcement communities is emphasized to create a more positive regulatory environment for pain relief and palliative care, which ultimately will benefit patient care. PMID- 15814512 TI - Continuous morphine infusions for cancer pain in resource-scarce environments: comparison of the subcutaneous and intravenous routes of administration. AB - Acute onset of severe pain in cancer patients may be due to multiple causes. Irrespective of the etiology, adequate analgesia has to be provided as quickly as possible. The standard practices of relieving pain by using syringe pumps (syringe drivers) or infusion pumps may not be feasible in resource-scarce developing nations where many cancer patients first present at advanced stages of disease for management. This study compared the efficacy of the subcutaneous and intravenous routes of morphine administration continuously using a simple and economic technique for cancer pain management. Both routes were found to be equally effective in producing good analgesia without side effects. The drip method is a cost-effective way of providing subcutaneous morphine infusion for cancer patients and is applicable for both inpatients and home care. PMID- 15814513 TI - Levetiracetam as an adjunctive analgesic in neoplastic plexopathies: case series and commentary. AB - Certain types of pain associated with cancer may be difficult to treat with standard therapies, often resulting in intractable pain and suffering for the patient. The use of an opioid as analgesic monotherapy can lead to poorly controlled pain as well as multiple side effects. Non-opioid adjunctive analgesics, such as antidepressants and antiepileptic drugs (AEDs) often improve both pain control, and side effect prevalence. Levetiracetam is an AED with unique mechanisms of action that may have analgesic properties in various pain syndromes. Seven patients with neoplasms involving neural structures (four invading the brachial plexus, and three the lumbosacral plexus) had severe pain of 8 to 9 out of 10 on a visual analog scale (VAS), despite the use of parenteral opioids and various adjunctive therapies. These patients were treated with oral levetiracetam titrated over days to two weeks, depending on the location of pain, drug response, and tolerance to tapering of opioid analgesics. Opioid and adjunctive analgesic use and VAS scores were recorded periodically. The maximum levetiracetam dose ranged from 500 mg to 1500 mg BID. All patients experienced pain control improvement after the addition of levetiracetam, with VAS scores decreasing from 8-9 out of 10 to 0-3 out of 10 within two to 14 days of therapy initiation. Overall opioid use decreased by at least an estimated 70%, without drug related adverse events. In this small series of patients, levetiracetam effectively and safely improved pain relief in patients with neoplastic plexopathies previously resistant to standard analgesic approaches. PMID- 15814514 TI - Patient-controlled analgesia safety issues. AB - Patient controlled analgesia (PCA) offers meaningful advantages over traditional analgesia. However, PCA medical errors with PCA can be dangerous and even fatal. Errors associated with patient controlled analgesia (PCA) therapy are described as are ways to minimize such errors. This report is based on data from the United States Pharmacopeia (USP) and Institute for Safe Medication practices (ISMP) Medication Errors Reporting Program. PCA by proxy, appropriate patients selection for PCA, patient monitoring, patient education, medication mix ups and practice related errors are discussed. Staff training is addressed. Methods to minimize errors associated with pump purchasing, PCA prrescribing, initiation, and monitoring are described. Recommendations for pharmaceutical manufacturers elated to PCA therapy are provided. PMID- 15814515 TI - Opioid side effects, addiction, and anti-inflammatory medications. AB - Patients in pain often fear medications prescribed or recommend to them by their clinicians. Fear of side effects can contribute greatly to medication non adherence (noncompliance). Patients often have fears that exceed the potential problems with which their medications are associated. Questions and answers relating to the side effects and the risk of addiction associated with opioids are presented. PMID- 15814516 TI - Cost-utility analysis and quality adjusted life years. AB - Cost utility analysis is a form of cost-effectiveness analysis in which outcomes are adjusted for quality and quantity of life. This type of analysis is used widely in Europe and is being used increasingly in the United States. This article provides an overview of cost utility analysis and quality adjusted life years, a commonly used effectiveness measure in CUA when comparing two or more treatments or interventions. PMID- 15814517 TI - Evidence-based pain management and palliative care in issue three for 2004 of The Cochrane Library. AB - The Cochrane Library of Systematic Reviews is published quarterly. Issue three for 2004 of the library was published in July 2004. The Cochrane Library now contains 3559 complete reviews and protocols of reviews. It also contains one page summaries of systematic reviews published in the general (non-Cochrane) medical literature. In addition there are citations of randomized controlled trials, methodology reviews, the Cochrane methodology register and the health technology assessment database. This edition of the Library contains the results of an extensive search for randomized controlled trials (RCTs) on EMBASE. The latest library contains 75 new reviews, five of which are considered relevant to practitioners in pain and palliative care. References are published in the same format as the citation for Cochrane reviews. PMID- 15814518 TI - Maximum intravenous morphine push rate: discrepancies between the primary and tertiary literature. PMID- 15814519 TI - More on current issues in pain management for the primary care practitioner. Acute pain: a multi-modal management approach. PMID- 15814520 TI - Prescription pain medications: frequently asked questions and answers for health care professionals, and law enforcement personnel. AB - A consensus document on frequently asked questions, answers and references relating to the use and abuse o f prescription analgesics was released to the public in August 2004 by the Drug Enforcement Administration, Last Acts Partnership, and the Pain & Policies Study Group of the University of Wisconsin. This document provides clear answers to common questions and is intended for primary care clinicians and drug enforcement and regulatory personnel. It clearly states that police and regulatory personnel should not interfere with medical practice and that clinicians have an obligation to practice in a legal manner. Topics addressed include risk assessment, how opioid treatment works, patient behavior, abuse, addiction, rules and laws, and clear descriptions of how and why the DEA may prosecute a clinician. PMID- 15814524 TI - Hepatitis B virus DNA-negative dane particles lack core protein but contain a 22 kDa precore protein without C-terminal arginine-rich domain. AB - DNA-negative Dane particles have been observed in hepatitis B virus (HBV) infected sera. The capsids of the empty particles are thought to be composed of core protein but have not been studied in detail. In the present study, the protein composition of the particles was examined using new enzyme immunoassays for the HBV core antigen (HBcAg) and for the HBV precore/core proteins (core related antigens, HBcrAg). HBcrAg were abundant in fractions slightly less dense than HBcAg and HBV DNA. Three times more Dane-like particles were observed in the HBcrAg-rich fraction than in the HBV DNA-rich fraction by electron microscopy. Western blots and mass spectrometry identified the HBcrAg as a 22-kDa precore protein (p22cr) containing the uncleaved signal peptide and lacking the arginine rich domain that is involved in binding the RNA pregenome or the DNA genome. In sera from 30 HBV-infected patients, HBcAg represented only a median 10.5% of the precore/core proteins in enveloped particles. These data suggest that most of the Dane particles lack viral DNA and core capsid but contain p22cr. This study provides a model for the formation of the DNA-negative Dane particles. The precore proteins, which lack the arginine-rich nucleotide-binding domain, form viral RNA/DNA-negative capsid-like particles and are enveloped and released as empty particles. PMID- 15814525 TI - Dangerous severe personality disorders: England's experiment in using psychiatry for public protection. PMID- 15814526 TI - Breastfeeding prevalence and practices among Singaporean Chinese, Malay and Indian mothers. AB - The National Breastfeeding Survey 2001 was the first comprehensive study on breastfeeding conducted on a national level in Singapore. It aimed to establish the prevalence of breastfeeding among Chinese, Malay and Indian mothers and to identify factors influencing breastfeeding. A total of 2098 mothers were interviewed in this two-phase study, with the first interview conducted 2 months after delivery and the second interview 6 months after birth among mothers who were still breastfeeding at 2 months. Frequency distributions of breastfeeding prevalence and types of breastfeeding practices at different time intervals (from birth to 6 months) were produced. Multivariate logistic regression was carried out to construct a model with predictive information on factors which influence continued breastfeeding till 2 months and 6 months after delivery respectively. The study found that about 94.5% of the mothers attempted breastfeeding. At 1 month, 71.6% were still breastfeeding, 49.6% continued to do so at 2 months, and 29.8% persisted till 4 months. By 6 months, the breastfeeding prevalence rate fell to 21.1%. The results of this study show higher breastfeeding prevalence rates compared to past studies in Singapore. Despite this, exclusive breastfeeding is still not a common practice. Various factors were found to be significant in influencing mothers' decision to breastfeed. Factors such as ethnicity, age, educational attainment, religion and baby's sex are non modifiable in the short term or at an individual level. However, factors such as awareness of breastfeeding benefits, advice from health professionals and previous breastfeeding experience are potentially modifiable. Efforts aimed at promoting breastfeeding in Singapore need to take these modifiable factors into consideration so as to better tailor health promotion efforts on breastfeeding to women. PMID- 15814527 TI - Evaluating the progress of the health promoting hospitals initiative? A WHO perspective. Commentary on: Whitehead, D. (2004) The European Health Promoting Hospitals (HPH) project: how far on? Health Promotion International, 19, 259-267. PMID- 15814528 TI - Exaggerated response to adenosine in kidneys from high salt-fed rats: role of epoxyeicosatrienoic acids. AB - Cytochrome P-450 (CYP)-dependent epoxyeicosatrienoic acids (EETs) dilate rat preglomerular microvessels when adenosine(2A) receptors (A(2A)R) are stimulated. As high salt (HS) intake increases epoxygenase activity and adenosine levels, we hypothesized that renal adenosine responses would be greater in HS-fed rats. Male Sprague-Dawley rats were fed either HS (4.0% NaCl) or normal salt (NS; 0.4% NaCl) diet. On day 8, isolated kidneys were perfused with Krebs' buffer containing indomethacin (10 microM) and L-NAME (200 microM) and preconstricted to approximately 150 mmHg with infusion of phenylephrine (10(-7) M). Renal effluents were extracted for analysis of eicosanoids by gas chromatography-mass spectrometry. Bolus injections of the stable adenosine analog 2-chloroadenosine (2-CA; 0.1-10 microg) resulted in dose-dependent dilation; at 10 microg, perfusion pressure (PP) was lowered to a greater extent in the kidneys of HS rats compared with NS rats (-60 +/- 4 vs. -31 +/- 8 mmHg; P < 0.05) and the area of response was increased (27 +/- 6 vs. 9 +/- 4 mm(2); P < 0.05), as was EET release (132 +/- 23 vs. 38 +/- 18 ng; P < 0.05). HS treatment increased A(2A)R and CYP2C23 protein expression. A selective epoxygenase inhibitor, MS-PPOH (12 microM), significantly reduced the response to 2-CA in HS rats; PP, area of response, and EET release decreased by 40, 70, and 81%, respectively, whereas lesser changes were evident in NS kidneys. Thus the greater vasodilator response to 2-CA seen in kidneys obtained from HS-fed rats was mediated by increased EET release. As EETs are renal vasodilator and natriuretic eicosanoids, interactions between adenosine and EETs may contribute to the adaptive response to HS intake. PMID- 15814529 TI - Glycation of mitochondrial proteins from diabetic rat kidney is associated with excess superoxide formation. AB - Chronic hyperglycemia causes structural alterations of proteins through the Maillard reaction. In diabetes, methylglyoxal (MGO)-induced hydroimidazolones are the predominant modification. In contrast to acute hyperglycemia, mitochondrial respiration is depressed in chronic diabetes. To determine whether MGO-derived protein modifications result in abnormalities in mitochondrial bioenergetics and superoxide formation, proteomics and functional studies were performed in renal cortical mitochondria isolated from rats with 2, 6, and 12 mo of streptozotocin induced diabetes. MGO-modified proteins belonged to the following two pathways: 1) oxidative phosphorylation and 2) fatty acid beta-oxidation. Two of these proteins were identified as components of respiratory complex III, the major site of superoxide production in health and disease. Mitochondria from rats with diabetes exhibited a diminution of oxidative phosphorylation. A decrease in the respiratory complex III activity was significantly correlated with the quantity of MGO-derived hydroimidazolone present on mitochondrial proteins in both diabetic and control animals. In diabetes, isolated renal mitochondria produced significantly increased quantities of superoxide and showed evidence of oxidative damage. Administration of aminoguanidine improved mitochondrial respiration and complex III activity and decreased oxidative damage to mitochondrial proteins. Therefore, posttranslational modifications of mitochondrial proteins by MGO may represent pathogenic events leading to mitochondria-induced oxidative stress in the kidney in chronic diabetes. PMID- 15814530 TI - Nedd4-2 isoforms differentially associate with ENaC and regulate its activity. AB - Mutations that disrupt a PY motif in epithelial Na(+) channel (ENaC) subunits increase surface expression of Na(+) channels in the collecting duct, resulting in greater Na(+) reabsorption. Nedd4 and Nedd4-2 have been identified as ubiquitin ligases that can interact with ENaC via its PY motifs to regulate channel activity. We recently reported that human Nedd4-2 (hNedd4-2) is expressed as many isoforms because of alternative promoter usage and/or variable splicing. To understand the relevance of hNedd4-2 isoforms for collecting duct Na(+) transport, we studied the interaction with ENaC and the intracellular localization and function of the following three naturally occurring hNedd4-2 isoforms: full-length Nedd4-2 (Nedd4-2), Nedd4-2 lacking the NH(2)-terminal C2 domain (Nedd4-2DeltaC2), and Nedd4-2 lacking the C2 domain and WW domains 2 and 3 (Nedd4-2DeltaWW2,3). Nedd4-2 and Nedd4-2DeltaC2 associate with ENaC and robustly reduce Na(+) transport in Xenopus oocytes, whereas the interaction with and functional effect of Nedd4-2DeltaWW2,3 on ENaC is weak. Nedd4-2 is expressed in the mouse collecting duct, and overexpression of Nedd4-2 reduces endogenous ENaC activity in a collecting duct cell line. This reduction in ENaC activity can be reversed early with exposure to dexamethasone, an effect that is associated with an increase in sgk1 abundance. The C2 domain is required to target Nedd4-2 to the plasma membrane in response to elevation of intracellular Ca(2+) concentration ([Ca(2+)](i)) in MDCK cells, although it does not appear to mediate the inhibitory effect of [Ca(2+)](i) on Na(+) transport. Our data illustrate that naturally occurring hNedd4-2 isoforms differentially associate with ENaC to regulate its activity. PMID- 15814531 TI - Regulation of the renal thiazide-sensitive Na-Cl cotransporter, blood pressure, and natriuresis in obese Zucker rats treated with rosiglitazone. AB - Previously, we showed an increase in protein abundance of the renal thiazide sensitive Na-Cl cotransporter (NCC) in young, prediabetic, obese Zucker rats relative to lean age mates (Bickel CA, Verbalis JF, Knepper MA, and Ecelbarger CA. Am J Physiol Renal Physiol 281: F639-F648, 2001). To test whether this increase correlated with increased thiazide sensitivity (NCC activity) and blood pressure, and could be modified by insulin-sensitizing agents, we treated lean and obese Zucker rats (9 wk old) with either a control diet or this diet supplemented with 3 mg/kg body wt rosiglitazone (RGZ), a peroxisomal proliferator activated receptor subtype gamma agonist and potent insulin-sensitizing agent, for 12 wk (n = 9/group). The rise in blood pressure, measured continuously by radiotelemetry, was significantly blunted in the RGZ-treated obese rats. Similarly, blood glucose and urinary albumin were markedly decreased in these rats. RGZ-treated rats whether lean or obese excreted a NaCl load faster but excreted less sodium in response to hydrochlorothiazide, applied as a novel in vivo measure of NCC activity. Obese rats had increased renal protein abundance and urinary excretion of NCC; however, this was not significantly reduced by RGZ (densitometry in cortex homogenate - %lean control): 100 +/- 9, 93 +/- 4, 124 +/- 9, and 141 +/- 14 for lean control, lean RGZ, obese control, and obese RGZ, respectively. Subcellular localization, as evaluated by confocal microscopy and immunoblotting following differential centrifugation, of NCC was not different between rat groups. Overall, RGZ reduced blood pressure and thiazide sensitivity; however, the mechanism(s) did not seem to involve a decrease in NCC protein abundance or cellular location. Decreased NCC activity may have contributed to the maintenance of normotension in RGZ-treated obese rats. PMID- 15814532 TI - Anti-inflammatory effect of fibrate protects from cisplatin-induced ARF. AB - Recently, we demonstrated that peroxisome proliferator-activated receptor-alpha (PPARalpha) ligand ameliorates cisplatin-induced acute renal failure (ARF) by preventing inhibition of substrate oxidation, and also by preventing apoptosis and necrosis of the proximal tubule (Li S, Bhatt R, Megyesi J, Gokden N, Shah SV, and Portilla D. Am J Physiol Renal Physiol 287: F990-F998, 2004). In the following studies, we examined the protective effect of PPARalpha ligand on cisplatin-induced inflammatory responses during ARF. Mice subjected to a single intraperitoneal injection of cisplatin developed ARF at day 3. Cisplatin increased mRNA and protein expression of TNF-alpha, RANTES, and also upregulated endothelial adhesion molecules ICAM-1/VCAM-1 and chemokine receptors CCR1/CCR5. Cisplatin also led to neutrophil infiltration in the corticomedullary region. Pretreatment of wild-type mice with WY-14,643, a fibrate class of PPARalpha ligands, before cisplatin significantly suppressed cisplatin-induced upregulation of cytokine/chemokine expression, prevented neutrophil accumulation, and ameliorated renal dysfunction. In contrast, treatment with PPARalpha ligand before cisplatin did not prevent cytokine/chemokine production, neutrophil accumulation, and did not protect kidney function in PPARalpha null mice. In addition, we observed that cisplatin-induced NF-kappaB binding activity in nuclear extracts from wild-type mice was markedly reduced by treatment with PPARalpha ligand. These results demonstrate that PPARalpha exerts an anti inflammatory effect in kidney tissue by a mechanism that includes inhibition of NF-kappaB DNA binding activity, and this effect results in inhibition of neutrophil infiltration, cytokine/chemokine release, and amelioration of cisplatin-induced ARF. PMID- 15814533 TI - Glucose degradation products downregulate ZO-1 expression in human peritoneal mesothelial cells: the role of VEGF. AB - BACKGROUND: Glucose degradation products (GDPs) are formed during heat sterilization of peritoneal dialysis fluid and, to a lesser extent, during their prolonged storage. In vitro studies have demonstrated that GDPs impair functions of peritoneal mesothelial cells, including proliferation, viability and cytokine release. In the present study, we studied the acute effect of GDPs on the expression of tight junction-associated protein, zonula occludens protein 1 (ZO 1), in human peritoneal mesothelial cells (HPMC). The role of the vascular endothelial growth factor (VEGF) induced by GDPs in the expression of ZO-1 was also examined. METHODS: HPMC were cultured with GDPs, including 2-furaldehyde (FurA), methylglyoxal (M-Glx) and 3,4-dideoxyglucosone-3-ene (3,4-DGE). The expression of ZO-1 and the synthesis of VEGF were examined. To define the role of VEGF on the regulation of ZO-1 expression, HPMC were cultured with GDPs in the presence or absence of neutralizing antibody to VEGF. The signal pathways involved in VEGF synthesis induced by GDPs were also characterized. RESULTS: ZO-1 expression in HPMC was downregulated in a time- and dose-dependent manner following culture with subtoxic concentrations of GDPs (FurA, M-Glx and 3,4-DGE). All three GDPs increased VEGF synthesis in HPMC. Exogenous VEGF downregulated the expression of ZO-1 and neutralizing anti-VEGF antibody reversed the effect of GDPs on ZO-1 expression in HPMC, suggesting the action of GDPs on ZO-1 expression was mediated by VEGF. All three GDPs activated the p42/p44 mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) signal transduction pathways. The GDP-induced VEGF and transforming growth factor (TGF)-beta synthesis in HPMC was partially reduced by either the p42/p44 MAPK inhibitor (PD98059) or the PKC inhibitor (staurosporine). More importantly, the VEGF and TGF-beta synthesis induced by GDPs in HPMC was completely blocked by synergistic action of both inhibitors. CONCLUSIONS: We have demonstrated that short-term exposure to GDPs downregulates ZO-1 expression in HPMC through the generation of VEGF. Our study provides evidence that GDPs can directly induce VEGF and TGF-beta production in HPMC through the activation of p42/44 MAPK and PKC signal transduction pathways. PMID- 15814534 TI - Chronic kidney disease as cause of cardiovascular morbidity and mortality. AB - To make an evidence-based evaluation of the relationship between kidney failure and cardiovascular risk, we reviewed the literature obtained from a PubMed search using pre-defined keywords related to both conditions and covering 18 years (1986 until end 2003). Eighty-five publications, covering 552 258 subjects, are summarized. All but three studies support a link between kidney dysfunction and cardiovascular risk. More importantly, the association is observed very early during the evolution of renal failure: an accelerated cardiovascular risk appears at varying glomerular filtration rate (GFR) cut-off values, which were >/=60 ml/min in at least 20 studies. Many studies lacked a clear definition of cardiovascular disease and/or used a single determination of serum creatinine or GFR as an index of kidney function, which is not necessarily corresponding to well-defined chronic kidney disease. In six studies, however, chronic kidney dysfunction and cardiovascular disease were well defined and the results of these confirm the impact of kidney dysfunction. It is concluded that there is an undeniable link between kidney dysfunction and cardiovascular risk and that the presence of even subtle kidney dysfunction should be considered as one of the conditions necessitating intensive prevention of this cardiovascular risk. PMID- 15814536 TI - Liver disease vs systemic inflammation in haemodialysis patients. PMID- 15814535 TI - Glucose-containing peritoneal dialysis fluids regulate leptin secretion from 3T3 L1 adipocytes. AB - BACKGROUND: A marked elevation of serum leptin is observed soon after the start of peritoneal dialysis (PD), suggesting that leptin production may be stimulated by this treatment. Glucose metabolism is the major factor regulating leptin. The current study was designed to test if glucose-based PD fluids might regulate leptin production in vitro. METHODS: 3T3-L1 adipocytes were exposed to a 50:50 mixture of dialysis solutions and medium M199 containing 10% serum for 15%). Secondary causes of small kidney were excluded. Of the seven cases, four had reduced renal function (67 vs 7% in the entire cohort), and three developed hyperparathyroidism during follow-up (43 vs 4%). The pathogenesis of SK ought to explain why anatomical structures of different embryological origin are involved (the precalyceal and collecting ducts and the nephron) and why there is frequent association with hyperparathyroidism. In embryogenesis, the metanephric blastema synthesizes the chemotactic glial derived neurotrophic factor (GDNF) to prompt the ureteric bud to branch off from Wolff's mesonephric duct, and to approach and invade the blastema. The bud's tip expresses the GDNF receptor (RET). RET-GDNF binding is crucial not only for the correct formation of ureters and collecting ducts (both of Wolffian origin), but also for nephrogenesis. We advance the hypothesis that SK results from a disruption in the ureteric bud-metanephric blastema interface, possibly due to one or more mutations or polymorphisms of RET or GDNF genes. This would explain: the concurrent alterations in precalyceal ducts and the functional defects in the nephron, the occasional association with size and the functional asymmetry between the two kidneys, some degree of renal dysplasia causing the reduction in the glomerular filtration rate and (given the role of RET in parathyroid cell proliferation) the association with hyperparathyroidism. PMID- 15814541 TI - Tailoring high-cut-off membranes and feasible application in sepsis-associated acute renal failure: in vitro studies. AB - BACKGROUND: As removal of pro-inflammatory cytokines is limited in conventional diffusive or convective extracorporeal therapies, we studied in two polysulphone membranes with an industrial albumin sieving coefficient of 0.05 (Type A) and 0.13 (Type B) cytokine (IL-6, IL-8, IL-1beta, IL-1ra, TNF-alpha) and plasma protein (albumin, cystatin C, total proteins) permeability profiles. Based on the convective membrane permeability, we evaluated in vitro the dialytic modality that could provide an acceptable balance between high cytokine and low albumin clearances. METHODS: Cytokine and plasma protein sieving coefficient (SC) and clearance were studied in (i) post-dilutional haemofiltration mode at 20% fixed ultrafiltration rate; (ii) haemodialysis mode (dialysate flow rate of 3 and 5 l/h); and (iii) haemodiafiltration mode (dialysate flow rate of 3 or 5 l/h with 0.5 l/h of ultrafiltrate). RESULTS: In haemofiltration mode both Type A and Type B haemodialysers at QB 150 ml/min exhibited similar median SC nearly up to 1 for IL-1beta and IL-1ra, at about 0.6 for IL-6, 0.4 for IL-8 and 0.7 for TNF-alpha, with clearance values ranging from 15 to 30 ml/min. SC were independent of blood flow and were stable throughout the whole experiment. Albumin SC was higher in Type B than in Type A and rapidly decreased from 0.2 to 0.02 and from 0.5 to 0.04 within 3 h for haemodialyser Types A and B, respectively. Cytokine SC was lower in haemodialysis than in haemodiafiltration and haemofiltration mode, and by increasing dialysate flow from 3 up to 5 l/h in both haemodialysis and haemodiafiltration mode, SC for all tested cytokines decreased. However, at 5 l/h clearances were not different or were higher, since increased amounts of dialysate outlet compensated for the decreased SC. Albumin clearances in haemodialysis and haemodiafiltration mode after 360 min at 5 l/h were 0.81 and 0.91 ml/min, respectively. CONCLUSIONS: Our studies show that a mixed convective and diffusive technique ensures high cytokine clearances with an acceptable loss of albumin. PMID- 15814542 TI - Myeloperoxidase serves as a marker of oxidative stress during single haemodialysis session using two different biocompatible dialysis membranes. AB - BACKGROUND: There is increased oxidative stress in patients undergoing haemodialysis (HD); however, little is known of how different dialysis membranes contribute to the oxidative stress induced by the dialysis procedure per se. We therefore studied the influence of two different dialysis membranes on oxidative stress during HD. METHODS: Eight patients undergoing HD three times per week were enrolled in this cross-controlled study. Patients sequentially received HD using polysulphone (PS) and regenerated cellulose (RC) dialysis membranes for 1 week each. Blood samples were collected in the last section of each hollow fibre 0, 15, 120 and 240 min after starting HD. We determined superoxide anion production derived from neutrophils, superoxide dismutase (SOD) and glutathione peroxidase (GPx) derived from washed red cells, plasma myeloperoxidase (MPO), plasma thiobarbituric acid-reactive substances (TBARS), plasma advanced oxidation protein products (AOPP) and serum 8-hydroxy-2'-deoxyguanosine (8-OHdG). RESULTS: Leukocyte numbers, including neutrophils, lymphocytes and monocytes, decreased significantly after 15 min of dialysis, more so with RC than with PS membrane. For both membranes, superoxide anion production transiently increased during the first 15 min whereas the post-dialysis production was decreased. Plasma MPO levels persistently increased during dialysis with the two membranes. Moreover, the increase was more marked with RC than with PS membrane. AOPP and 8-OHdG levels increased progressively when using RC membranes. There were no significant differences in SOD, GPx, TBARS, AOPP and 8-OHdG levels between the two membranes. CONCLUSIONS: The biocompatibility of the dialyser affects oxidative stress production during a single dialysis session. The measurement of MPO may serve as a reliable marker of the degree of oxidative stress induced using dialysis membranes of different biocompatibilities. PMID- 15814544 TI - Rejection after simultaneous pancreas-kidney transplantation. AB - BACKGROUND: Simultaneous pancreas-kidney (SPK) transplantation is an accepted therapy for type 1 diabetic patients with end-stage renal disease. This study analyses the occurrence of rejection episodes in patients undergoing SPK. METHODS: The study population was obtained from 205 patients enrolled in the Euro SPK 001 study and randomized to receive tacrolimus- (n = 103) or cyclosporin microemulsion (ME)-based (n = 102) immunosuppressive therapy. All patients received concomitant antibody induction therapy, mycophenolate mofetil and short term corticosteroids. RESULTS: After 3 years of follow-up, rejection episodes occurred in 41 patients receiving tacrolimus and in 51 patients receiving cyclosporin-ME. The majority of first rejection episodes in both groups occurred during the first 6 months (93 and 90%, respectively) and in most cases were treated with corticosteroids alone (88 vs 90%). Actuarial rejection-free kidney and/or pancreas graft survival was similar for tacrolimus (54%) and cyclosporin ME (44%). Human leukocyte antigen (HLA) compatibility (P = 0.003) and graft vessel extension (P = 0.000001) had a significant influence on rejection-free graft survival. Also, rejection influenced pancreas graft survival (P = 0.01), and pancreas graft loss due to rejection influenced patient survival (P = 0.02). In the intent-to-treat analysis of early rejection, significantly fewer tacrolimus- than cyclosporin-ME-treated patients had (i) more than one rejection episode (11 out of 40 vs 24 out of 47; P = 0.03); (ii) first moderate to severe rejection (one out of 40 vs 12 out of 47; P = 0.004); and (iii) refractory rejection (two out of 40 vs 10 out of 47; P = 0.03). Pancreas survival was lower in late rejectors (53%) than non-rejectors (86%; P = 0.002). Also, serum creatinine was highest in late rejectors. CONCLUSION: Tacrolimus-based immunosuppressive therapy demonstrates significant advantages over cyclosporin-ME in terms of the severity of acute rejection in SPK transplant patients. PMID- 15814545 TI - Metabolic results 3 years after simultaneous pancreas-kidney transplantation. AB - BACKGROUND: Simultaneous pancreas-kidney (SPK) transplantation has become accepted therapy for patients with type 1 diabetes and end-stage renal disease. This 3-year study compared the metabolic effects of tacrolimus- and cyclosporin microemulsion (ME)-based immunosuppressive therapy in this clinical setting. METHODS: The study population comprised the 205 patients enrolled in the Euro-SPK 001 study. Glucose metabolism parameters [fasting blood glucose, fasting C peptide and glycated haemoglobin (HbA1c)], blood lipids (total cholesterol and triglycerides) and pancreatic enzymes (lipase and amylase) were monitored at regular intervals during the study. Blood pressure was also carefully monitored and compared with target levels for diabetic patients. RESULTS: Fasting C-peptide and HbA1c levels were within the normal ranges in the two treatment groups throughout the 3 years. Fasting blood glucose was higher during the first 2 months post-transplant in the tacrolimus group than in the cyclosporin-ME group, but no differences were seen thereafter. From month 2 post-transplant, mean levels of total cholesterol were significantly lower among patients receiving tacrolimus than among those in the cyclosporin-ME group. In addition, patients receiving cyclosporin-ME showed serological features of mild pancreatitis, with elevated blood amylase and lipase levels during the first 6 months post transplant. The two regimens were comparable with respect to hypertension. CONCLUSIONS: Except for lipid profiles, no major differences in metabolic effects or blood pressure control were observed over the 3 years in SPK transplant patients receiving immunosuppression based on tacrolimus or cyclosporin-ME. In view of the potential risk of hypertension, antihypertensive strategies should be implemented for all patients. PMID- 15814546 TI - Cytomegalovirus: occurrence, severity, and effect on graft survival in simultaneous pancreas-kidney transplantation. AB - BACKGROUND: This analysis of the Euro-SPK 001 study examined the occurrence and effect of cytomegalovirus (CMV) infection during the first 3 years after simultaneous pancreas-kidney (SPK) transplantation. METHODS: In this multicentre study, 205 SPK transplant patients were randomized to immunosuppressive treatment with tacrolimus (n = 103) or cyclosporin microemulsion [(ME), n = 102]. All patients received antibody induction therapy, mycophenolate mofetil and short term corticosteroids. The choice of CMV prophylaxis and treatment was at the discretion of each investigator. RESULTS: The overall incidence of CMV infection was 34%, with equal distribution in the tacrolimus and cyclosporin-ME groups. Fewer CMV infections occurred with ganciclovir (22%) than aciclovir (43% P = 0.007) or no prophylaxis (42%, P = 0.008). The rates of CMV infection according to donor and recipient CMV serological status were: D-/R- 11%; D-/R+ (40%, P = 0.004); D+/R+ (37%, P = 0.002); and D+/R- (52%, P<0.001). In the three at-risk subgroups, infection rates were lower among patients receiving ganciclovir (22%) than among those receiving aciclovir or no prophylaxis (64%; P<0.0001). Acute rejection was more common among CMV-infected patients (66 vs 41% without infection, P = 0.001) and in those not receiving ganciclovir prophylaxis. The 3 year actuarial rejection-free survival rate was 61.4% with ganciclovir and 42.2% with no prophylaxis or aciclovir alone (P = 0.002). No differences were observed in actuarial patient, kidney or pancreas survival between CMV and non-CMV infection groups. CONCLUSIONS: Our findings confirm that the incidence of CMV infection is the same in tacrolimus- and cyclosporin-ME-treated SPK recipients. Ganciclovir prophylaxis effectively prevented CMV infection, especially in higher risk groups, and was associated with a reduced incidence of rejection compared with aciclovir/no prophylaxis. PMID- 15814547 TI - Efficacy and safety of tacrolimus compared with cyclosporin microemulsion in primary SPK transplantation: 3-year results of the Euro-SPK 001 trial. AB - BACKGROUND: Single-centre and retrospective studies suggest superiority of tacrolimus over cyclosporin as cornerstone immunosuppressive therapy for simultaneous pancreas-kidney (SPK) transplantation. This open-label, multicentre trial compared the efficacy and safety of tacrolimus with cyclosporin microemulsion (ME) in diabetic patients with end-stage renal disease undergoing their first cadaveric SPK transplantation. The 3-year results are reported. METHODS: Patients were recruited from 10 centres in Europe and one centre in Israel: 103 were randomized to receive tacrolimus (initial dose: 0.2 mg/kg/day p.o.) and 102 to cyclosporin-ME (7 mg/kg/day p.o.). All patients received concomitant rabbit anti-T-cell globulin induction, mycophenolate mofetil (MMF) and short-term corticosteroids. RESULTS: Fewer patients receiving tacrolimus (36.9%) than cyclosporin-ME (57.8%) were discontinued from treatment (P = 0.003). The initial episodes of biopsy-proven rejection were moderate or severe in just one out of 31 (3%) tacrolimus-treated patients compared with 11 out of 39 (28%) patients receiving cyclosporin-ME (P = 0.009). While 3-year patient and kidney survival rates were similar in the two treatment groups, pancreas survival was superior with tacrolimus (89.2 vs 72.4%; P = 0.002). Thrombosis resulted in pancreas graft loss in 10 patients receiving cyclosporin-ME and in only two treated with tacrolimus (P = 0.02). Overall adverse event frequency was similar in both groups, but MMF intolerance was more frequent with tacrolimus and hyperlipidaemia more frequent with cyclosporin-ME. CONCLUSIONS: In this 3-year study, tacrolimus was more effective than cyclosporin-ME in preventing moderate or severe kidney or pancreas rejection after SPK transplantation. It also provided superior pancreas survival and reduced the risk of pancreas graft thrombosis. PMID- 15814548 TI - Secondary effects of immunosuppressive drugs after simultaneous pancreas-kidney transplantation. AB - BACKGROUND: This report examines the early and late secondary effects of tacrolimus, cyclosporin microemulsion (ME), mycophenolate mofetil (MMF) and rabbit anti-thymocyte globulin (rATG) in simultaneous pancreas-kidney (SPK) recipients. METHODS: Of the 205 patients participating in the Euro-SPK 001 study, 103 were randomized to tacrolimus (0.2 mg/kg) and 102 to cyclosporin-ME (7 mg/kg). All patients received rATG for 4 days [ATG-Fresenius (ATG-F) 4 mg/kg/day or Thymoglobulin (Thymo-S) 1.25 mg/kg/day] plus MMF and short-term corticosteroids. RESULTS: Thymo-S induction therapy was associated with a lower white cell count in the first 3 months than was seen with ATG-F, while ATG-F caused a lower initial nadir in platelet count. Both polyclonal preparations were well tolerated and no clinically relevant differences were observed with respect to side effects such as infections and malignancies. High cyclosporin-ME trough levels were associated with pancreas graft thrombosis, and concentrations >150 ng/ml were associated with poor renal allograft function. Treatment discontinuation was higher with cyclosporin-ME (57.8%) than with tacrolimus-based therapy (36.9%) due to more frequent toxicity, graft loss and lack of efficacy requiring a switch to tacrolimus. The main reason for withdrawal in the tacrolimus group was MMF discontinuation; MMF-related side effects resulted in more frequent dose reductions to <2 g/day and discontinuations in the tacrolimus group, and indirectly indicate a higher dose-corrected exposure to mycophenolic acid, as previously observed in kidney transplant patients. CONCLUSIONS: Short term induction therapy is effective and well tolerated in patients undergoing SPK transplantation. Tacrolimus was the preferred immunosuppressive agent, resulting in fewer cases of pancreas graft loss and drug discontinuation compared with cyclosporin-ME. PMID- 15814549 TI - A large, prospective, randomized, open-label, multicentre study of corticosteroid withdrawal in SPK transplantation: a 3-year report. AB - BACKGROUND: Simultaneous pancreas-kidney (SPK) transplantation is the treatment of choice for selected diabetic patients. Corticosteroids are an important element of immunosuppressive protocols, but their long-term use has detrimental effects on patients' health, necessitating eventual discontinuation. METHODS: This prospective study evaluated the safety and feasibility of corticosteroid withdrawal in 205 SPK transplant recipients randomized to immunosuppressive treatment with either tacrolimus and mycophenolate mofetil (MMF) (n = 103) or cyclosporin microemulsion (ME) and MMF (n = 102). RESULTS: Corticosteroid withdrawal was successful in the majority of in-study patients (66% tacrolimus, 73% cyclosporin-ME). Compared with out-of-study patients or those continuing corticosteroid therapy, in-study patients withdrawn from corticosteroids experienced fewer pancreas or kidney graft losses, fewer episodes of acute rejection and were less likely to be withdrawn from the study. Acute rejection occurred after corticosteroid withdrawal in two patients who had a previous rejection and in five patients who were rejection-free before corticosteroid withdrawal. No rejection episodes were associated with graft loss or immediate serious consequences. Overall, corticosteroid withdrawal was achieved with an increase in the dose of both MMF and tacrolimus. CONCLUSIONS: A long-term survey of corticosteroid withdrawal in SPK transplantation with multifactorial analyses is necessary to confirm these early results and to evaluate the positive effects on glucose metabolism and hypertension. PMID- 15814550 TI - Impact of HLA matching on the outcome of simultaneous pancreas-kidney transplantation. AB - BACKGROUND: Simultaneous pancreas-kidney (SPK) transplantation has become the therapy of choice for type 1 diabetic patients with end-stage renal disease. The current analysis examined the impact of human leukocyte antigen (HLA) matching on graft outcome following SPK transplantation. The study population was obtained from patients enrolled in the Euro-SPK 001 study. METHODS: The effect of HLA matching on graft function and survival was assessed in 180 SPK recipients in whom complete donor-recipient HLA data were available. A group of 45 patients with 0-3 HLA mismatches (MM) was compared with a group of 135 patients with 4-6 MM. RESULTS: There were no differences in 3-year kidney, pancreas or patient survival between the 0-3 and 4-6 MM groups. Biological parameters of kidney and pancreas graft function were similar in both groups. Significantly more patients with 0-3 MM (66%) were rejection-free at 3 years than was the case among those with 4-6 MM (41%; P = 0.003). The relative risk of acute rejection was 2.6 times higher among patients with 4-6 MM than among those with 0-3 MM. CONCLUSIONS: There was no evidence that HLA matching was associated with improved kidney or pancreas survival. However, a higher rate of acute rejection was observed with poor HLA match, which may impact long-term survival. PMID- 15814551 TI - Spectrum of surgical complications after simultaneous pancreas-kidney transplantation in a prospectively randomized study of two immunosuppressive protocols. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation (SPK) has evolved as an effective treatment for patients with end-stage nephropathy due to type 1 diabetes mellitus. This report analyses the spectrum of surgical complications among patients receiving tacrolimus and cyclosporin microemulsion (ME)-based therapy for SPK transplantation. METHODS: The analysis included 205 patients randomly assigned to tacrolimus (n = 103) or cyclosporin-ME (n = 102) in the Euro SPK 001 study. Surgical complications were defined as any intervention in the 3 month post-operative course related to the transplant procedure. RESULTS: In the tacrolimus vs cyclosporin-ME group, repeat laparotomy was required by fewer patients (26 vs 43%, respectively; P = 0.01) and at a later stage post-transplant (26+/-26 vs 14+/-17 days; P = 0.05). Also, thrombosis of graft vessels (2 vs 9%; P = 0.03) and repeat laparotomy for intra-abdominal haemorrhage within the first 3 months (8 vs 22%; P = 0.005) occurred significantly less frequently with tacrolimus vs cyclosporin-ME. A donor age of > or =45 years was a significant determinant for surgical complications requiring repeat laparotomy, regardless of the type of immunosuppression. Portal anastomosis was the safest method of endocrine venous drainage, and Roux-en-Y loop for enteric exocrine drainage was associated with a higher re-operation rate than duodenoenterostomy. Repeat laparotomy had no impact on patient survival, but significantly reduced kidney and pancreas graft survival in the cyclosporin-ME group (kidney: P<0.01; pancreas: P<0.001) and in both groups combined (P < or = 0.05 and P<0.001, respectively). CONCLUSIONS: The immunological benefits of tacrolimus compared with cyclosporin-ME treatment result in a lower incidence of repeat laparotomies post-transplant and a reduced in-hospital stay. Fewer repeat laparotomies translate into improved pancreas and kidney graft survival. PMID- 15814553 TI - BEST: binding-site estimation suite of tools. AB - SUMMARY: The purpose of our Binding-site Estimation Suite of Tools (BEST) is two fold: to provide a platform for using and comparing different motif-finding programs for transcription factor binding site prediction, and to improve the accuracy of these predictions by further optimization. Our software package BEST includes four commonly used motif-finding programs: AlignACE, BioProspector, CONSENSUS and MEME, as well as the optimization program BioOptimizer. BEST allows the user to run programs either separately or sequentially and manages all programs by automating the common inputs and the optimization procedure. The BEST system was implemented in Qt, a C++ application development framework, and was compiled and executed on Linux operating systems. AVAILABILITY: BEST is available for download at http://www.cs.uga.edu/~che/BEST and http://www.fas.harvard.edu/~junliu/BEST CONTACT: dsche@uga.edu, jliu@stat.harvard.edu. PMID- 15814554 TI - EGene: a configurable pipeline generation system for automated sequence analysis. AB - EGene is a generic, flexible and modular pipeline generation system that makes pipeline construction a modular job. EGene allows for third-party programs to be used and integrated according to the needs of distinct projects and without any previous programming or formal language experience being required. EGene comes with CoEd, a visual tool to facilitate pipeline construction and documentation. A series of components to build pipelines for sequence processing is provided. AVAILABILITY: http://www.lbm.fmvz.usp.br/egene/ CONTACT: alan@ime.usp.br; argruber@usp.br SUPPLEMENTARY INFORMATION: http://www.lbm.fmvz.usp.br/egene/ PMID- 15814555 TI - Prediction of protein solvent accessibility using fuzzy k-nearest neighbor method. AB - MOTIVATION: The solvent accessibility of amino acid residues plays an important role in tertiary structure prediction, especially in the absence of significant sequence similarity of a query protein to those with known structures. The prediction of solvent accessibility is less accurate than secondary structure prediction in spite of improvements in recent researches. The k-nearest neighbor method, a simple but powerful classification algorithm, has never been applied to the prediction of solvent accessibility, although it has been used frequently for the classification of biological and medical data. RESULTS: We applied the fuzzy k-nearest neighbor method to the solvent accessibility prediction, using PSI BLAST profiles as feature vectors, and achieved high prediction accuracies. With leave-one-out cross-validation on the ASTRAL SCOP reference dataset constructed by sequence clustering, our method achieved 64.1% accuracy for a 3-state (buried/intermediate/exposed) prediction (thresholds of 9% for buried/intermediate and 36% for intermediate/exposed) and 86.7, 82.0, 79.0 and 78.5% accuracies for 2-state (buried/exposed) predictions (thresholds of each 0, 5, 16 and 25% for buried/exposed), respectively. Our method also showed slightly better accuracies than other methods by about 2-5% on the RS126 dataset and a benchmarking dataset with 229 proteins. AVAILABILITY: Program and datasets are available at http://biocom1.ssu.ac.kr/FKNNacc/ CONTACT: jul@ssu.ac.kr. PMID- 15814556 TI - Penalized Cox regression analysis in the high-dimensional and low-sample size settings, with applications to microarray gene expression data. AB - MOTIVATION: An important application of microarray technology is to relate gene expression profiles to various clinical phenotypes of patients. Success has been demonstrated in molecular classification of cancer in which the gene expression data serve as predictors and different types of cancer serve as a categorical outcome variable. However, there has been less research in linking gene expression profiles to the censored survival data such as patients' overall survival time or time to cancer relapse. It would be desirable to have models with good prediction accuracy and parsimony property. RESULTS: We propose to use the L(1) penalized estimation for the Cox model to select genes that are relevant to patients' survival and to build a predictive model for future prediction. The computational difficulty associated with the estimation in the high-dimensional and low-sample size settings can be efficiently solved by using the recently developed least-angle regression (LARS) method. Our simulation studies and application to real datasets on predicting survival after chemotherapy for patients with diffuse large B-cell lymphoma demonstrate that the proposed procedure, which we call the LARS-Cox procedure, can be used for identifying important genes that are related to time to death due to cancer and for building a parsimonious model for predicting the survival of future patients. The LARS-Cox regression gives better predictive performance than the L(2) penalized regression and a few other dimension-reduction based methods. CONCLUSIONS: We conclude that the proposed LARS-Cox procedure can be very useful in identifying genes relevant to survival phenotypes and in building a parsimonious predictive model that can be used for classifying future patients into clinically relevant high- and low risk groups based on the gene expression profile and survival times of previous patients. PMID- 15814557 TI - Multiclass cancer classification and biomarker discovery using GA-based algorithms. AB - MOTIVATION: The development of microarray-based high-throughput gene profiling has led to the hope that this technology could provide an efficient and accurate means of diagnosing and classifying tumors, as well as predicting prognoses and effective treatments. However, the large amount of data generated by microarrays requires effective reduction of discriminant gene features into reliable sets of tumor biomarkers for such multiclass tumor discrimination. The availability of reliable sets of biomarkers, especially serum biomarkers, should have a major impact on our understanding and treatment of cancer. RESULTS: We have combined genetic algorithm (GA) and all paired (AP) support vector machine (SVM) methods for multiclass cancer categorization. Predictive features can be automatically determined through iterative GA/SVM, leading to very compact sets of non redundant cancer-relevant genes with the best classification performance reported to date. Interestingly, these different classifier sets harbor only modest overlapping gene features but have similar levels of accuracy in leave-one-out cross-validations (LOOCV). Further characterization of these optimal tumor discriminant features, including the use of nearest shrunken centroids (NSC), analysis of annotations and literature text mining, reveals previously unappreciated tumor subclasses and a series of genes that could be used as cancer biomarkers. With this approach, we believe that microarray-based multiclass molecular analysis can be an effective tool for cancer biomarker discovery and subsequent molecular cancer diagnosis. PMID- 15814558 TI - MEDS and PocR are novel domains with a predicted role in sensing simple hydrocarbon derivatives in prokaryotic signal transduction systems. AB - We identify two conserved domains in diverse bacterial and archaeal signaling proteins. One of them, the MEDS domain, is typified by the DmcR protein from Methylococcus and the other by the PocR protein of Salmonella typhi. We provide evidence that both these domains are likely to sense simple hydrocarbon derivatives and transduce downstream signals on binding these ligands. The PocR ligand-binding domain is shown to contain a novel variant of the fold found in PAS and GAF domains. The MEDS domain is present in both methylotrophs and complex methanogens, and both the MEDS and PocR domains show a lineage-specific expansion in the latter organisms, suggesting a role in sensing their principle growth substrates. The MEDS domain is also found in the negative regulators of the sigma factor SigB in actinomycetes, including pathogens like Mycobacterium tuberculosis. Hence it is possible that these sigma factors, involved in aerial mycelium development and stress response in the actinomycetes, might be under the regulation of as yet uncharacterized small molecules. CONTACT: aravind@ncbi.nlm.nih.gov. PMID- 15814559 TI - A model-based scan statistic for identifying extreme chromosomal regions of gene expression in human tumors. AB - MOTIVATION: The analysis of gene expression data in its chromosomal context has been a recent development in cancer research. However, currently available methods fail to account for variation in the distance between genes, gene density and genomic features (e.g. GC content) in identifying increased or decreased chromosomal regions of gene expression. RESULTS: We have developed a model-based scan statistic that accounts for these aspects of the complex landscape of the human genome in the identification of extreme chromosomal regions of gene expression. This method may be applied to gene expression data regardless of the microarray platform used to generate it. To demonstrate the accuracy and utility of this method, we applied it to a breast cancer gene expression dataset and tested its ability to predict regions containing medium-to-high level DNA amplification (DNA ratio values >2). A classifier was developed from the scan statistic results that had a 10-fold cross-validated classification rate of 93% and a positive predictive value of 88%. This result strongly suggests that the model-based scan statistic and the expression characteristics of an increased chromosomal region of gene expression can be used to accurately predict chromosomal regions containing amplified genes. AVAILABILITY: Functions in the R language are available from the author upon request. CONTACT: fcouples@umich.edu. PMID- 15814560 TI - Common denominator procedure: a novel approach to gene-expression data mining for identification of phenotype-specific genes. AB - MOTIVATION: We have established a novel data mining procedure for the identification of genes associated with pre-defined phenotypes and/or molecular pathways. Based on the observation that these genes are frequently expressed in the same place or in close proximity at about the same time, we have devised an approach termed Common Denominator Procedure. One unusual feature of this approach is that the specificity and probability to identify genes linked to the desired phenotype/pathway increase with greater diversity of the input data. RESULT: To show the feasibility of our approach, the Cancer Genome Anatomy Project expression data combined with a defined set of angiogenic factors was used to identify additional and novel angiogenesis-associated genes. A multitude of these additional genes were known to be associated with angiogenesis according to published data, verifying our approach. For some of the remaining candidate genes, application of a high-throughput functional genomics platform (XantoScreen) provided further experimental evidence for association with angiogenesis. PMID- 15814561 TI - POSBIOTM-NER: a trainable biomedical named-entity recognition system. AB - SUMMARY: POSBIOTM-NER is a trainable biomedical named-entity recognition system. POSBIOTM-NER can be automatically trained and adapted to new datasets without performance degradation, using CRF (conditional random field) machine learning techniques and automatic linguistic feature analysis. Currently, we have trained our system on three different datasets. GENIA-NER was trained based on GENIA Corpus, GENE-NER based on BioCreative data and GPCR-NER based on our own POSBIOTM/NE corpus, respectively, which would be used in GPCR-related pathway extraction. PMID- 15814562 TI - Probe rank approaches for gene selection in oligonucleotide arrays with a small number of replicates. AB - MOTIVATION: One major area of interest in analyzing oligonucleotide gene array data is identifying differentially expressed genes. A challenge to biostatisticians is to develop an approach to summarizing probe-level information that adequately reflects the true expression level while accounting for probe variation, chip variation and interaction effects. Various statistical tools, such as MAS and RMA, have been developed to address this issue. In these approaches, the probe level expression data are summarized into gene level data, which are then used for downstream statistical analysis. Since probe variation is often larger than chip variation and there is also a potential interaction effect between probe affinity and treatment effect, strategies such as a gene level analysis, may not be optimal. In this study, we propose a procedure to analyze probe level data for selecting differentially expressed genes under two treatment conditions (groups) with a small number of replicates. The probe level discrepancy between two groups can be measured by a difference of the percentiles of probe perfect-match (PM) ranks or of probe PM weighted ranks. The difference is then compared with a pre-specified threshold to determine differentially expressed genes. The probe level approach takes into account non-homogenous treatment effects and reduces possible cross-hybridization effects across a set of probes. RESULTS: The proposed approach is compared with MAS and RMA using two benchmark gene array datasets. Positive predictivity and sensitivity are used for evaluation. Results show the proposed approach has higher positive predictivity and higher sensitivity. AVAILABILITY: Available on request from the authors. CONTACT: dtchen@uab.edu. PMID- 15814563 TI - ROBIN: a tool for genome rearrangement of block-interchanges. AB - SUMMARY: ROBIN is a web server for analyzing genome rearrangement of block interchanges between two chromosomal genomes. It takes two or more linear/circular chromosomes as its input, and computes the number of minimum block-interchange rearrangements between any two input chromosomes for transforming one chromosome into another and also determines an optimal scenario taking this number of rearrangements. The input can be either bacterial-size sequence data or landmark-order data. If the input is sequence data, ROBIN will automatically search for the identical landmarks that are the homologous/conserved regions shared by all the input sequences. PMID- 15814564 TI - VariScan: Analysis of evolutionary patterns from large-scale DNA sequence polymorphism data. AB - SUMMARY: VeriScan is a software package for the analysis of DNA sequence polymorphisms at the whole genome scale. Among other features, the software (1) can conduct many population genetic analyses; (2) incorporates a multiresolution wavelet transform-based method that allows capturing relevant information from DNA polymorphism data; (3) facilitates the visualization of the results in the most commonly used genome browsers. PMID- 15814565 TI - Literature mining and database annotation of protein phosphorylation using a rule based system. AB - MOTIVATION: A large volume of experimental data on protein phosphorylation is buried in the fast-growing PubMed literature. While of great value, such information is limited in databases owing to the laborious process of literature based curation. Computational literature mining holds promise to facilitate database curation. RESULTS: A rule-based system, RLIMS-P (Rule-based LIterature Mining System for Protein Phosphorylation), was used to extract protein phosphorylation information from MEDLINE abstracts. An annotation-tagged literature corpus developed at PIR was used to evaluate the system for finding phosphorylation papers and extracting phosphorylation objects (kinases, substrates and sites) from abstracts. RLIMS-P achieved a precision and recall of 91.4 and 96.4% for paper retrieval, and of 97.9 and 88.0% for extraction of substrates and sites. Coupling the high recall for paper retrieval and high precision for information extraction, RLIMS-P facilitates literature mining and database annotation of protein phosphorylation. PMID- 15814566 TI - Long-term outcome of patients with heart failure and dynamic functional mitral regurgitation. AB - AIMS: In patients with heart failure and chronic ischaemic mitral regurgitation (MR), the mortality risk is related to the quantified severity of MR at rest and its dynamic changes during exercise. The impact of dynamic MR on long-term mortality, hospital admission for heart failure, and major adverse cardiac events has never been investigated. METHODS AND RESULTS: We prospectively studied 161 patients with chronic ischaemic left ventricular (LV) dysfunction and at least mild MR who underwent quantitative measurement of the effective regurgitant orifice (ERO) of MR at rest and during semi-supine exercise test and who were followed up for 35+/-11 months. The 20 patients who underwent surgery were censored at the time of operation. Of the 141 patients who were treated medically, 23 died, 22 required hospitalization for heart failure, 4 had nonfatal myocardial infarction, and 11 developed unstable angina. By multivariate analysis, an exercise-induced increase in ERO by > or =13 mm(2) and a greater increase in transtricuspid pressure gradient during exercise emerged as predictors of mortality and of hospital admission for heart failure. MR severity under basal conditions (ERO> or =20 mm(2)) was an independent predictor of only cardiac death. Greater LV volumes at rest and lack of contractile reserve during exercise were additional independent markers of poor outcome. CONCLUSION: In patients with ischaemic heart disease and LV dysfunction, large exercise-induced increases in MR identify patients at high risk of morbidity and of death. PMID- 15814567 TI - A new definition for myocardial infarction: what difference does it make? AB - AIMS: As a response to changing diagnostic tools of myocardial infarction (MI), new case definitions for acute coronary events were published in 2003 as the American Heart Association Scientific Statement. We assessed the new definition in hospitalized patients in a large population-based MI register study. METHODS AND RESULTS: We identified all suspected acute coronary syndromes with data either on troponin T or on troponin I and at least one of the enzymatic markers of myocardial injury (n=6104). The 2003 definition with the use of troponins identified 83% more definite MIs than the WHO MONICA definition using cardiac enzymes. The additional patients were older, had more often diabetes, and received less often thrombolysis and revascularization than those having MI by both definitions. Adjusting for age, sex, study area, and study year, the additional patients with their first MI aged 25-74 had a higher risk of cardiovascular death within 1 year than patients having definite MI by both definitions (hazard ratio 1.6, 95% CI 1.1-2.2). CONCLUSION: The changing diagnostic criteria present a considerable challenge for the assessment of long term trends in MI events in the community as well as for longitudinal studies of the natural history of MI. The 2003 definition, when applied using troponins, identified a sizable new group of MI patients, among persons with suspected acute coronary syndrome, at high risk of a recurrent event. PMID- 15814568 TI - Inhibition of ribonucleotide reductase reduces neointimal formation following balloon injury. AB - Percutaneous transluminal coronary angioplasty (PTCA) has greatly benefited patients with occluded coronary arteries, but its benefits have been undermined by a high incidence of restenosis. The introduction of coronary stents has significantly improved the short and long term outcome but restenosis still occurs in approximately 15 to 30% of patients within 6 months. Research efforts are now being directed toward combination stenting and drug delivery. Among the therapeutic targets being pursued are agents that can impede smooth muscle cell migration and proliferation, as these processes are critical components of restenosis injury. We propose that inhibiting the conversion of ribonucleotides to deoxyribonucleotides will impede cell proliferation and, as such, limit the degree of restenosis. Therefore, we tested whether the potent ribonucleotide reductase inhibitors Didox (3,4-dihydroxybenzohydraxamic acid) and Imidate (ethyl 3,4,5-hydroxybenzimidate) can limit the neointimal proliferation associated with restenosis using a rat carotid model of balloon dilatation injury. Results demonstrated that both Didox and Imidate significantly reduced intimal thickening, resulting in a 71 and 62% decrease in the intima/media ratio, respectively. Similar efficacy was seen with the commercially available ribonucleotide reductase inhibitor hydroxyurea, demonstrating the importance of this enzyme in vascular remodeling. Results from cell proliferation studies suggest that the mechanism of protection is inhibition of smooth muscle cell (SMC) proliferation. In addition, Didox and Imidate (100 microM) are potent inhibitors of SMC migration, which may also contribute to their vascular protective effects. These results suggest that inhibition of ribonucleotide reductase may provide a potent strategy to prevent post-PTCA restenosis. PMID- 15814569 TI - Anxiolytic effects of Maxipost (BMS-204352) and retigabine via activation of neuronal Kv7 channels. AB - Neuronal Kv7 channels are recognized as potential drug targets for treating hyperexcitability disorders such as pain, epilepsy, and mania. Hyperactivity of the amygdala has been described in clinical and preclinical studies of anxiety, and therefore, neuronal Kv7 channels may be a relevant target for this indication. In patch-clamp electrophysiology on cell lines expressing Kv7 channel subtypes, Maxipost (BMS-204352) exerted positive modulation of all neuronal Kv7 channels, whereas its R-enantiomer was a negative modulator. By contrast, at the Kv7.1 and the large conductance Ca2+-activated potassium channels, the two enantiomers showed the same effect, namely, negative and positive modulation at the two channels, respectively. At GABA(A) receptors (alpha1beta2gamma2s and alpha2beta2gamma2s) expressed in Xenopus oocytes, BMS-204352 was a negative modulator, and the R-enantiomer was a positive modulator. The observation that the S- and R-forms exhibited opposing effects on neuronal Kv7 channel subtypes allowed us to assess the potential role of Kv7 channels in anxiety. In vivo, BMS 204352 (3-30 mg/kg) was anxiolytic in the mouse zero maze and marble burying models of anxiety, with the effect in the burying model antagonized by the R enantiomer (3 mg/kg). Likewise, the positive Kv7 channel modulator retigabine was anxiolytic in both models, and its effect in the burying model was blocked by the Kv7 channel inhibitor 10,10-bis-pyridin-4-ylmethyl-10H-anthracen-9-one (XE-991) (1 mg/kg). Doses at which BMS-204352 and retigabine induce anxiolysis could be dissociated from effects on sedation or memory impairment. In conclusion, these in vitro and in vivo studies provide compelling evidence that neuronal Kv7 channels are a target for developing novel anxiolytics. PMID- 15814570 TI - Molecular determinants of picrotoxin inhibition of 5-hydroxytryptamine type 3 receptors. AB - Previously, we reported that the GABA(A) receptor antagonist picrotoxin also antagonizes serotonin (5-HT)3 receptors and that its effects are subunit dependent. Here, we sought to identify amino acids involved in picrotoxin inhibition of 5-HT3 receptors. Mutation of serine to alanine at the transmembrane domain 2 (TM2) 2' position did not affect picrotoxin (PTX) sensitivity in murine 5-HT3A receptors. However, mutation of the 6' TM2 threonine to phenylalanine dramatically reduced PTX sensitivity. Mutation of 6' asparagine to threonine in the 5-HT3B subunit enhanced PTX sensitivity in heteromeric 5-HT3A/3B receptors. Introduction of serine (native to the human 3B subunit) at the 6' position also increased PTX sensitivity, suggesting a species-specific effect. Mutation of the 7' leucine to threonine in 5-HT3A receptors increased PTX sensitivity roughly 10 fold, comparable with that observed in GABA(A) receptors, and also conferred distinct gating kinetics. The equivalent mutation in the 3B subunit (i.e., 7' valine to threonine) had no impact on PTX sensitivity in 5-HT3A/3B receptors. Interestingly, [3H]ethynylbicycloorthobenzoate ([3H]EBOB), a high-affinity ligand to the convulsant site in GABA(A) receptors, did not exhibit specific binding in 5-HT3A receptors. The structurally related compound, tert butylbicyclophosphorothionate (TBPS), which potently inhibits GABA(A) receptors, did not inhibit 5-HT3 currents. Our results indicate that the TM2 6' residue is a common determinant of PTX inhibition of both 5-HT3 and GABA(A) receptors and demonstrate a role of the 7' residue in PTX inhibition. However, lack of effects of EBOB and TBPS in 5-HT3A receptors suggests that the functional domains in the two receptors are not equivalent and underscores the complexity of PTX modulation of LGICs. PMID- 15814571 TI - Analysis of the in vivo functions of Mrp3. AB - Multidrug resistance protein 3 (MRP3) is an ATP-binding cassette transporter that is able to confer resistance to anticancer agents such as etoposide and to transport lipophilic anions such as bile acids and glucuronides. These capabilities, along with the induction of the MRP3 protein on hepatocyte sinusoidal membranes in cholestasis and the expression of MRP3 in enterocytes, have led to the hypotheses that MRP3 may function in the body to protect normal tissues from etoposide, to protect cholestatic hepatocytes from endobiotics, and to facilitate bile-acid reclamation from the gut. To elucidate the role of Mrp3 in these processes, the Mrp3 gene (Abcc3) was disrupted by homologous recombination. Homozygous null animals were healthy and physically indistinguishable from wild-type mice. Mrp3(-/-) mice did not exhibit enhanced lethality to etoposide phosphate, although an analysis of transfected human embryonic kidney 293 cells indicated that the potency of murine Mrp3 toward etoposide ( approximately 2.0- to 2.5-fold) is comparable with that of human MRP3. After induction of cholestasis by bile duct ligation, Mrp3(-/-) mice had 1.5-fold higher levels of liver bile acids and 3.1-fold lower levels of serum bilirubin glucuronide compared with ligated wild-type mice, whereas significant differences were not observed between the respective sham-operated mice. Bile acid excretion, pool size, and fractional turnover rates were similar in Mrp3(-/ ) and wild-type mice. We conclude that Mrp3 functions as an alternative route for the export of bile acids and glucuronides from cholestatic hepatocytes, that the pump does not play a major role in the enterohepatic circulation of bile acids and that the lack of chemosensitivity is probably attributable to functional redundancy with other pumps. PMID- 15814572 TI - GABAA receptor-associated protein regulates GABAA receptor cell-surface number in Xenopus laevis oocytes. AB - GABA(A) receptor-associated protein (GABARAP) was isolated previously in a yeast two-hybrid screen using the intracellular loop of the gamma2 subunit of the GABA(A) receptor as bait. GABARAP has been shown to participate in the membrane clustering and intracellular-trafficking of GABA(A) receptors, including a stimulation of the surface expression of GABA(A) receptors. To assess this quantitatively, we used Xenopus laevis oocytes expressing alpha1beta2gamma2S containing GABA(A) receptors to demonstrate that coexpression of GABARAP increased net surface levels of GABA(A) receptors as shown by both increased GABA currents and surface-expressed protein. This GABARAP stimulation of GABA currents required the receptor gamma2 subunit and full-length GABARAP: deletion of the microtubule-binding domain (amino acids 1-22) or disrupting the polymerization of microtubules abolished the enhancement, indicating that the effect of GABARAP was derived from the interaction with microtubules. GABARAP coexpression did not alter the general properties of GABA(A) receptors such as sensitivity to GABA or benzodiazepines, but it increased surface levels of receptor protein in oocytes. Rather, it seems to supplement inadequate amounts of endogenous GABARAP to support optimum trafficking and/or stabilization of surface GABA(A) receptors. PMID- 15814573 TI - Possible involvement of CPI-17 in augmented bronchial smooth muscle contraction in antigen-induced airway hyper-responsive rats. AB - Airway hyper-responsiveness (AHR) associated with heightened airway resistance and inflammation is a characteristic feature of asthma. It has been demonstrated that contractile responsiveness and Ca(2+) sensitization to acetylcholine (ACh) in repeated antigen challenge-induced airway hyper-responsive bronchial preparation were significantly increased. The CPI-17 (PKC-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase of 17 kDa) is activated by protein kinase C and acts on a myosin light-chain phosphatase-specific target. The aim of the present study was to explore the role of CPI-17 in hyper responsiveness of bronchial smooth muscle in antigen-induced AHR rats. In immunoblotting, the levels of expression of CPI-17 mRNA and protein were significantly increased in bronchus from rats that were repeatedly challenged with antigen. ACh-induced CPI-17 phosphorylation and translocation to membrane fraction were also significantly increased in bronchus from antigen-challenged rats. In conclusion, we suggest that augmented expression and activation of CPI 17 observed in the hyper-responsive bronchial smooth muscle might be responsible for the enhanced ACh-induced Ca(2+) sensitization of bronchial smooth muscle contraction associated with AHR. PMID- 15814574 TI - Fatal progressive systemic sclerosis following autologous stem cell transplantation and high-dose chemotherapy. PMID- 15814575 TI - Relationship between rheumatoid arthritis and Mycoplasma pneumoniae: a case control study. AB - OBJECTIVE: Rheumatoid arthritis (RA) has a complex and multifactorial aetiology. Infectious agents could start this disease. The majority of the characteristics of this infirmity can be observed in chronic arthritis produced by mycoplasmas in animals. In this study the association between Mycoplasma pneumoniae and RA has been evaluated. METHODS: A case-control study was performed. Sera taken from 78 RA patients and from 156 controls were analysed to ascertain the levels of immunoglobulin G (IgG) against M. pneumoniae. Other variables, like age, gender, work status, history of pneumonia, etc., were recorded in a questionnaire. RESULTS: The presence of antibodies against M. pneumoniae was associated with RA (odds ratio=2.34, P<0.001). CONCLUSIONS: The results suggest that M. pneumoniae could be a cofactor in the pathogenesis of RA; however, more studies need to be done. PMID- 15814576 TI - Neuroendocrine-immune relationships between emotion regulation and health in patients with rheumatoid arthritis. AB - OBJECTIVES: Emotion regulation is hypothesized to be related to health through neuroendocrine-immune changes. This study examined the role of the neuroendocrine variables 24-h urinary cortisol and noradrenaline, and the immune variable serum interleukin 6 as mediators between emotion regulation styles and health (perceived health and disease activity: erythrocyte sedimentation rate (ESR) and Thompson joint score). METHODS: Sixty patients with rheumatoid arthritis (mean age 59.0+/-11.2 yr; 38 female) participated. RESULTS: Emotion regulation was not associated with immune functioning or disease activity, but it was somewhat related to neuroendocrine functioning: one of the emotion regulation styles, ambiguity, was related to noradrenaline in women (r = 0.39) but not in men. The indicators of neuroendocrine functioning (cortisol and noradrenaline) were correlated (r = 0.40), as were indicators of immune functioning (interleukin 6) and inflammatory activity (ESR; r = 0.53), but analyses did not indicate a role of these physiological variables in mediating between emotion regulation and health: neuroendocrine variables were not related to interleukin 6 or ESR, and none of the physiological parameters was correlated with joint score or perceived health. CONCLUSIONS: To examine whether the proposed mediational processes apply to individual patients, a longitudinal within-subjects design is needed. In our cross-sectional study, emotion regulation was somewhat related to neuroendocrine functioning, but our study did not uncover a potential mediational role of cortisol, noradrenaline or interleukin 6 in the relationship between emotion regulation and health in rheumatoid arthritis. PMID- 15814577 TI - BILAG 2004. Development and initial validation of an updated version of the British Isles Lupus Assessment Group's disease activity index for patients with systemic lupus erythematosus. AB - OBJECTIVE: To devise a more discriminating version of the British Isles Lupus Assessment Group (BILAG) disease activity index and to show that it is reliable. METHODS: A nominal consensus approach was undertaken by members of BILAG to update and improve the BILAG lupus disease activity index. The index has been revised following intense consultations over a 1-yr period. It has been assessed in two real-patient exercises. These involved patients with diverse clinical features of SLE, including gastrointestinal, hepatic and ophthalmic problems, which the earlier versions of the index did not fully take into account. Reliability in terms of the ability to differentiate patients was assessed by calculating intraclass correlation coefficients. The level of agreement between physicians was determined by calculating the ratio of estimates of the standard error (SE) attributable to the physicians to the SE attributable to the patients. RESULTS: Good reliability and high levels of physician agreement were observed in one or both exercises in the constitutional, mucocutaneous, neurological, cardiorespiratory, renal, ophthalmic and haematological systems. In contrast, the musculoskeletal system did not score as well, although providing more clear-cut glossary definitions should greatly improve the situation. CONCLUSIONS: Some significant changes in the BILAG disease activity index to assess patients with SLE are proposed. The process of demonstrating validity and reliability has started with these two exercises assessing real patients. Further validation studies are under way. BILAG 2004 is likely to be valuable in clinical trials assessing new therapies for the treatment of SLE, as it provides a more comprehensive system-based disease activity measure than has been available previously. PMID- 15814578 TI - Genetic and genomic studies of PADI4 in rheumatoid arthritis. AB - OBJECTIVES: Strong genetic association of rheumatoid arthritis (RA) with PADI4 (peptidyl arginine deiminase) has previously been described in Japanese, although this was not confirmed in a subsequent study in the UK. We therefore undertook a further study of genetic association between PADI4 and RA in UK Caucasians and also studied expression of PADI4 in the peripheral blood of patients with RA. METHODS: Seven single-nucleotide polymorphisms (SNP) were genotyped using polymerase chain reaction (PCR)-restriction fragment length polymorphism in 111 RA cases and controls. A marker significantly associated with RA (PADI4_100, rs#2240339) in this first data set (P = 0.03) was then tested for association in a larger group of 439 RA patients and 428 controls. PADI4 transcription was also assessed by real-time quantitative PCR using RNA extracted from peripheral blood mononuclear cells from 13 RA patients and 11 healthy controls. RESULTS: A single SNP was weakly associated with RA (P = 0.03) in the initial case-control study, a single SNP (PADI4_100) and a two marker haplotype of that SNP and the neighbouring SNP (PADI4_104) were significantly associated with RA (P = 0.02 and P = 0.03 respectively). PADI4_100 was not associated with RA in a second sample set. PADI4 expression was four times greater in cases than controls (P = 0.004), but expression levels did not correlate with the levels of markers of inflammation. CONCLUSION: PADI4 is significantly overexpressed in the blood of RA patients but genetic variation within PADI4 is not a major risk factor for RA in Caucasians. PMID- 15814579 TI - Identifying patients with chronic kidney disease from general practice computer records. AB - BACKGROUND: Chronic kidney disease (CKD) is an important predictor of end-stage renal disease, as well as a marker of increased mortality. The New Opportunities for Early Renal Intervention by Computerised Assessment (NEOERICA) project aimed to assess whether people with undiagnosed CKD who might benefit from early intervention could be identified from GP computer records. METHODS: The simplified Modification of Diet in Renal Disease (MDRD) equation was used to estimate glomerular filtration rate (GFR) and determine stage of CKD in patients from 12 practices in Surrey, Kent and Greater Manchester with SCr recorded in their notes. Further data were extracted on associated co-morbidities and potentially modifiable risk factors. RESULTS: One quarter (25.7%; 28,862/112,215) had an SCr recorded and one in five (18.9%) of them had a GFR <60 ml/min/1.73 m2 (equivalent to Stage 3-5 CKD), representing 4.9% of the population. Only 3.6% of these were recorded as having renal disease. Three-quarters (74.6%; 4075/5449) of those with Stage 3-5 CKD had one or more circulatory diseases; 346 were prescribed potentially nephrotoxic drugs and over 4000 prescriptions were issued for drugs recommended to be used with caution in renal impairment. CONCLUSIONS: Patients with CKD can be identified by searching GP computer databases; along with associated co-morbidities and treatment. Results revealed a similar rate of Stage 3-5 CKD to that found previously in the USA. The very low rate of recording of renal disease in patients found to have CKD indicates scope for improving detection and early intervention. PMID- 15814580 TI - A brief case-finding questionnaire for common mental disorders: the CMDQ. AB - OBJECTIVES: The aim of the study was to validate a new case-finding instrument for common mental disorders (CMDQ). METHODS: A cross-sectional, stratified, two phase study was carried out in 28 general practices in Aarhus County, Denmark. 1785 consecutive patients, 18-65 years old, consulting 38 GPs with a new health problem participated. Patients were screened before consultation using a one-page screening questionnaire including subscales for somatisation (SCL-SOM and Whiteley-7), anxiety (SCL-ANX4), depression (SCL-DEP6) and alcohol abuse (CAGE). A stratified subsample of 701 patients was interviewed using the Schedules for Clinical Assessment in Neuropsychiatry (SCAN) interview. We tested the external validity of the scales using the SCAN interview as gold standard. All data were analysed using appropriate weighted procedures to control for the two-phase sampling design and non-response bias. RESULTS: Estimates of sensitivity and specificity for relevant ICD-10 diagnoses at theoretical optimal cut-off points on subscales: Depressive disorder: 78/86 (SCL-DEP6); Alcohol abuse or dependence: 78/97 (CAGE); Severe anxiety disorder: 77/85 (SCL-ANX4); Somatisation disorder: 83/56 (SCL-SOM); and 75/52 (Whiteley-7); any mental disorder: 72/72 (SCL-8). At the theoretical optimal cut-off points the CMDQ demonstrated higher diagnostic accuracy than GPs on any diagnosis evaluated. CONCLUSION: The study results suggest that the CMDQ has excellent external validity for use as a diagnostic aid in primary care settings. PMID- 15814581 TI - Many people with epilepsy want to know more: a qualitative study. AB - OBJECTIVE: To explore why, at the turn of the 21st century, many people with epilepsy still want more information. METHODS: Qualitative study with a maximum variation sample in the UK. We interviewed 38 men and women, 35 with epilepsy and three carers of people with epilepsy, recruited through GPs, neurologists, support groups and charities. RESULTS: People with epilepsy obtained information via health professionals, epilepsy organisations, leaflets, books, and the Internet. Many people wanted much more information about treatment options. People wanted to discuss the advantages and disadvantages of different drugs with their consultants, who were sometimes too busy. People also wanted to know more about the causes of epilepsy, partly because they feared the stigma associated with it, and other matters, such as Sudden Unexpected Death in Epilepsy [SUDEP]. CONCLUSION: The UK government plans to provide better services for those with epilepsy but, without enough money to employ more specialists, the information needs of patients may not be met. For concordance to work effectively doctors need to be aware of what patients think about their drugs, and patients need more information about treatment options. They also need more time to discuss the causes of epilepsy in order to reduce stigma and fear. Since time in consultations is short, clinicians should inform patients about epilepsy organisations, and direct them to websites such as the DIPEx (Personal Experiences of Health and Illness) epilepsy website (www.dipex.org/epilepsy), which focuses on patients' experiences of epilepsy and provides reliable information about medicines, other treatments, and resources. PMID- 15814582 TI - General practice as a complex system: a novel analysis of consultation data. AB - BACKGROUND: Complex systems have specific properties of robustness and self organisation which arise from interacting components within the overall system and which govern the system's behaviour. These are typically associated with a power law distribution of event sizes. Commentators have suggested that health systems are complex, but there has been limited quantitative investigation of this issue. OBJECTIVES: To test the hypothesis that consultation patterns in primary care follow a power law distribution typical of a complex system. METHODS: Analysis of 142,050 episodes of non-pathological back pain in routinely collected New Zealand national data. Calculation of the distribution of the duration and number of GP consultations for each illness episode. Secondary analysis of a published UK dataset of consultation rates for 44,000 patients in four general practices. RESULTS: Number of consultations per episode of back pain demonstrated excellent fit with a power law in the full dataset (r2 = 0.96) and all but one subgroups (r2 = 0.90-0.99). The number of consultations per patient from four UK practices was suggestive of a power law distribution (r2 = 0.88 0.93). CONCLUSIONS: Consultation patterns in general practice show measurable properties of a complex system. The consistency of the distribution across different population groups suggests that attempts to manage consultation patterns should focus on the whole system of patients, rather than upon individuals or subgroups of the patient population. PMID- 15814583 TI - Participation in a quality of care study and consequences for generalizability of general practice research. AB - BACKGROUND: Non-participation of General Practitioners (GPs) hampers primary care research. Using existing network structures can improve participation, but may introduce sampling effects. OBJECTIVES: To examine the role of network-based recruitment and other determinants of participation, and to estimate effects of sampling and non-participation on generalizability. METHODS: For a study of quality of care assessment, we recruited GPs from a regionally defined population and a GP network. Effects of sampling and non-participation were analysed by comparing characteristics between participants, target samples, and reference data for all German GPs. Factors influencing study participation were assessed in multiple logistic regression. RESULTS: Compared to the regional sample, network GPs were more likely to complete survey questionnaires (92% versus 69%) and to participate in the study (66% versus 23%). Compared to national reference data, study participants from both populations were younger, had a higher level of professional training, and included more men. These differences were already present in the network target sample, but were largely attributable to selective participation in the regional sample. Network membership remained the strongest determinant of participation in multiple logistic regression (odds ratio 5.01; 95% confidence interval 2.53-9.91). A younger age, higher professional training, and membership in the German Society of General Practice were also predictive of participation. CONCLUSIONS: Although network-based recruitment of GPs increases participation rates, sample effects are similar in size and direction as effects of non-participation in the regionally defined population. Careful analysis of participants based on publicly available data is therefore crucial for the assessment of generalizability. PMID- 15814584 TI - Costing in primary care--is the truth out there? PMID- 15814585 TI - Effect of preventive health screening on long-term primary health care utilization. A randomized controlled trial. AB - BACKGROUND: Regular preventive health screenings are a feature of primary health care in several countries. Studies of the effect of regular preventive health checks have reported different results regarding primary health care utilization. OBJECTIVE: To analyse the effect of preventive health screening and health discussions on contacts to general practice. METHODS: A randomized controlled trial with all GPs in the district of Ebeltoft, Denmark. All middle-aged residents registered with a GP in the district of Ebeltoft were included (n = 3464). A random sample of 2030 subjects was selected for invitation to participate in health screening or health screening and discussions. The remaining 1434 subjects were never contacted and served as an external control group. Main outcome measure was number of daytime consultations in general practice. RESULTS: The annual rate ratios for daytime consultations showed a very clear time trend (P < 0.0001) with a high rate of contacts among invited compared with non-invited subjects during the first year (P = 0.001) followed by a gradual decrease to a lower level after eight years (P = 0.037). The total rate ratio for daytime consultations was 1.01 (95% CI 0.93 to 1.10). CONCLUSION: We observed no differences between the invited group and the non-invited group in any type of contact to general practice when the entire follow-up period was considered. There was a significant trend in rate ratios for daytime consultations with an initial rise followed by a gradual decrease in rate ratios. More investigations are needed to confirm and explore reasons for this trend. PMID- 15814586 TI - Common chromosomal fragile site FRA16D mutation in cancer cells. AB - Neither the molecular basis for common fragile site DNA instability nor the contribution of this form of chromosomal instability to cancer is clearly understood. Fragile site FRA16D (16q23.2) is within regions of frequent loss-of heterozygosity (LOH) in breast and prostate cancers, is associated with homozygous deletions in various adenocarcinomas and t(14;16) chromosomal translocations in multiple myeloma. The FOR (WWOX) gene spans FRA16D and encodes a partner of p53 that also has a role in apoptosis. Previously untested 53 cancer cell lines were screened for deletions within the FOR/WWOX gene. Deletions were detected in Co115, KM12C and KM12SM. Homozygous deletions in these and two previously identified tumour cell lines were intragenic on both alleles, indicating a distinct mutation mechanism from that causing LOH. Identical FRA16D deletions in two cell lines (one derived from the primary carcinoma and the other from a secondary metastasis) demonstrate that FRA16D DNA instability can be an early, transient event. Sequence analysis across one deletion locates one endpoint within a polymorphic AT-dinucleotide repeat and the other adjacent to an AT-rich mini-satellite repeat implicating AT-rich repeats in FRA16D DNA instability. Another deletion is associated with de novo repetition of the 9 bp AT-rich sequence at one of the deletion endpoints. FRA16D deleted cells retain cytogenetic fragile site expression indicating that the deletions are susceptible sites for breakage rather than regions that confer fragility. Most cell lines with FRA16D homozygous deletions also have FRA3B deletions, therefore common fragile sites represent highly susceptible genome-wide targets for a distinct form of mutation. PMID- 15814587 TI - Shared gene expression profiles in individuals with autoimmune disease and unaffected first-degree relatives of individuals with autoimmune disease. AB - Patients with autoimmune disorders exhibit highly reproducible gene expression profiles in their peripheral blood mononuclear cells. These signatures may result from chronic inflammation, other disease manifestations, or may reflect family resemblance. To test the latter hypothesis, we determined gene expression profiles in unaffected first-degree relatives of individuals with autoimmune disease. Gene expression profiles in unaffected first-degree relatives resembled the profiles found in individuals with autoimmune diseases. A high percentage of differentially expressed genes in unaffected first-degree relatives were previously identified as autoimmune signature genes. Examination of the linear regression relationship of gene transcript levels between parent-offspring pairs revealed that autoimmune signature genes display high levels of family resemblance. Taken together, these results support the hypothesis that these variations in gene transcript levels are associated with family resemblance rather than clinical manifestations of disease. PMID- 15814588 TI - Regulation of elastin gene transcription by proteasome dysfunction. AB - Elastin, a major extracellular matrix protein and the core component of elastic fiber, is essential to maintain lung structural integrity and normal physiological function. We previously found that the downregulation of elastin gene transcription by IL-1beta is mediated via activation of NF-kappaB and CCAAT/enhancer binding protein (C/EBP)beta, both targets of the ubiquitin proteasome pathway. To further investigate the molecular mechanisms that underlie the control of elastin gene expression, we disrupted the ubiquitin-proteasome pathway with specific proteasome inhibitors. We found that specific proteasome inhibitors decreased the steady-state level of elastin mRNA in a dose-responsive manner. Run-on assay and promoter reporter study indicated that the proteasome inhibitor MG-132 repressed the rate of elastin transcription. MG-132 did not affect mRNA levels of NF-kappaB and C/EBPbeta, or the nuclear presence of NF kappaB, but markedly increased C/EBPbeta isoforms, including liver-enriched transcriptional activating protein and liver-enriched transcriptional inhibitory protein. Addition of cycloheximide blocked these increases and the downregulation of elastin mRNA by MG-132. The MG-132-induced downregulation of elastin transcription was dependent on C/EBPbeta expression as assessed with small interfering RNA. These results indicate that the ubiquitin-proteasome pathway plays an essential role in maintaining elastin gene expression in lung fibroblasts. Disruption of this pathway results in the downregulation of tropoelastin transcription via posttranscriptionally induced C/EBPbeta isoforms. PMID- 15814589 TI - PSGL-1 derived from human neutrophils is a high-efficiency ligand for endothelium expressed E-selectin under flow. AB - P-selectin glycoprotein ligand-1 (PSGL-1) has been proposed as an important tethering ligand for E-selectin and is expressed at a modest level on human leukocytes. Sialyl Lewis x (sLe(x))-like glycans bind to E-selectin and are expressed at a relatively high level on circulating leukocytes. It is unclear whether PSGL-1 has unique biochemical attributes that contribute to its role as an E-selectin ligand. To probe this issue, we conjugated microspheres with either sLe(x) or PSGL-1 purified from myeloid cells (neutrophils and HL-60) and compared their adhesion to endothelial expressed E-selectin under defined shear conditions. We found that both sLe(x) and PSGL-1 microspheres adhere to 4 h of IL 1beta-activated human umbilical vein endothelial cells predominantly through E selectin. Analysis of the adhesion revealed that the rate of initial tethering of the PSGL-1 microspheres to E-selectin was significantly greater than the rate of initial tethering of the sLe(x) microspheres despite the fact that the sLe(x) microspheres tested had higher ligand densities than the PSGL-1 microspheres. We also found that pretreatment of the PSGL-1 or sLe(x) microspheres with HECA-452 had no significant effect on initial tethering to E-selectin. These results support the hypotheses that 1) PSGL-1 is a high-efficiency tethering ligand for E selectin, 2) ligand biochemistry can significantly influence initial tethering to E-selectin, and 3) PSGL-1 tethering to E-selectin can occur via non-HECA-452 reactive epitopes. PMID- 15814590 TI - RhoA-Rho kinase pathway mediates thrombin- and U-46619-induced phosphorylation of a myosin phosphatase inhibitor, CPI-17, in vascular smooth muscle cells. AB - Protein kinase C-potentiated phosphatase inhibitor of 17 kDa (CPI-17) mediates some agonist-induced smooth muscle contraction by suppressing the myosin phosphatase in a phosphorylation-dependent manner. The physiologically relevant kinases that phosphorylate CPI-17 remain to be identified. Several previous studies have shown that some agonist-induced CPI-17 phosphorylation in smooth muscle tissues was attenuated by the Rho kinase (ROCK) inhibitor Y-27632, suggesting that ROCK is involved in agonist-induced CPI-17 phosphorylation. However, Y-27632 has recently been found to inhibit protein kinase C (PKC)-delta, a well-recognized CPI-17 kinase. Thus the role of ROCK in agonist-induced CPI-17 phosphorylation remains uncertain. The present study was designed to address this important issue. We selectively activated the RhoA pathway using inducible adenovirus-mediated expression of a constitutively active mutant RhoA (V14RhoA) in primary cultured rabbit aortic vascular smooth muscle cells (VSMCs). V14RhoA caused expression level-dependent CPI-17 phosphorylation at Thr38 as well as myosin phosphatase phosphorylation at Thr853. Importantly, we have shown that V14RhoA-induced CPI-17 phosphorylation was not affected by the PKC inhibitor GF109203X but was abolished by Y-27632, suggesting that ROCK but not PKC was involved. Furthermore, we have shown that the contractile agonists thrombin and U 46619 induced CPI-17 phosphorylation in VSMCs. Similarly to V14RhoA-induced CPI 17 phosphorylation, thrombin-induced CPI-17 phosphorylation was not affected by inhibition of PKC with GF109203X, but it was blocked by inhibition of RhoA with adenovirus-mediated expression of exoenzyme C3 as well as by Y-27632. Taken together, our present data provide the first clear evidence indicating that ROCK is responsible for thrombin- and U-46619-induced CPI-17 phosphorylation in primary cultured VSMCs. PMID- 15814591 TI - The abts and sulp families of anion transporters from Caenorhabditis elegans. AB - The slc4 and slc26 gene families encode two distinct groups of gene products that transport HCO(3)(-) and other anions in mammalian cells. The SLC4 and SLC26 proteins are important contributors to transepithelial movement of fluids and electrolytes and to cellular pH and volume regulation. Herein we describe the cDNA cloning from the nematode Caenorhabditis elegans of four anion bicarbonate transporter (abts) homologs of slc4 cDNA and eight sulfate permease (sulp) homologs of slc26 cDNA. Analysis of transgenic nematode strains carrying promoter::GFP fusions suggests relatively restricted expression patterns for many of these genes. At least three genes are expressed primarily in the intestine, three are expressed primarily in the excretory cell, and one is expressed in both of these polarized cell types. One of the genes is also expressed exclusively in the myoepithelium-like cells of the pharynx. Many of the sulp gene products localize to the basolateral membrane rather than to the apical membrane. Several ABTS and SULP proteins exhibited anion transport function in Xenopus oocytes. The strongest Cl(-) transporter among these also mediated Cl(-)/HCO(3)(-) exchange. These findings encourage exploitation of the genetic strengths of the nematode model system in the study of the physiological roles of anion transport by the proteins of these two highly conserved gene families. PMID- 15814592 TI - Long term cardiac follow up of severe twin to twin transfusion syndrome after intrauterine laser coagulation. AB - OBJECTIVE: To assess long term changes in cardiac morphology and function in survivors of severe twin to twin transfusion syndrome (TTTS) after intrauterine laser coagulation of placental anastomoses. DESIGN: Prospective follow up of fetuses with severe TTTS treated by laser coagulation of intrauterine placental anastomoses. Fetal echocardiography and Doppler studies of feto-placental haemodynamic function were performed at the time of laser coagulation (median gestational age of 21.7 weeks). Postnatal cardiac follow up included a detailed echocardiographic study of systolic and diastolic cardiac function at a median age of 21.1 months. SETTING: Paediatric cardiology unit. PATIENTS: 89 survivors from 73 consecutive pregnancies with severe TTTS. RESULTS: Before laser treatment, 28 of 51 (54.9%) recipient twins had typical signs of cardiac dysfunction due to volume overload and 9 of 38 (23.7%) donors had absent or reversed end diastolic flow in the umbilical artery. Echocardiography was normal in 87.6% of the survivors (34 of 38 donors, 44 of 51 recipients). The prevalence of congenital heart disease and particularly of pulmonary stenosis, which was recorded only in recipients, was increased in comparison with the general population (congenital heart disease, 10 of 89 (11.2%) v 0.3%; pulmonary stenosis, 4 of 51 (7.8%) v 0.03%). Findings before laser treatment were not correlated with the development of structural heart disease. CONCLUSIONS: Despite the high rate and severity of prenatal cardiac overload in recipients, the majority of cases of TTTS are normalised after laser treatment. However, given the increased prevalence of congenital heart disease and in particular pulmonary stenosis, intrauterine and postnatal follow up is warranted. PMID- 15814593 TI - Long term prognostic value of myocardial viability and ischaemia during dobutamine stress echocardiography in patients with ischaemic cardiomyopathy undergoing coronary revascularisation. AB - OBJECTIVE: To evaluate the relative merits of viability and ischaemia for prognosis after revascularisation. METHODS: Low-high dose dobutamine stress echocardiography (DSE) was performed before revascularisation in 128 consecutive patients with ischaemic cardiomyopathy (mean (SD) left ventricular ejection fraction (LVEF) 31 (8)%). Viability (defined as contractile reserve (CR)) and ischaemia were assessed during low and high dose dobutamine infusion, respectively. Cardiac death was evaluated during a five year follow up. Clinical, angiographic, and echocardiographic data were analysed to identify predictors of events. RESULTS: Univariable predictors of cardiac death were the presence of multivessel disease (hazard ratio (HR) 0.21, p < 0.001), baseline LVEF (HR 0.90, p < 0.0001), wall motion score index (WMSI) at rest (HR 4.02, p = 0.0006), low dose DSE (HR 7.01, p < 0.0001), peak dose DSE (HR 4.62, p < 0.0001), the extent of scar (HR 1.39, p < 0.0001), and the presence of CR in > or = 25% of dysfunctional segments (HR 0.34, p = 0.02). The best multivariable model to predict cardiac death included the presence of multivessel disease, WMSI at low dose DSE, and the presence of CR in > or = 25% of the severely dysfunctional segments (HR 9.62, p < 0.0001). Inclusion of ischaemia in the model did not provide additional predictive value. CONCLUSION: The findings of the present study illustrate that in patients with ischaemic cardiomyopathy, the extent of viability (CR) is a strong predictor of long term prognosis after revascularisation. Ischaemia did not add significantly in predicting outcome. PMID- 15814594 TI - Alteration of proximal aorta biophysical properties in patients with end stage renal disease. AB - OBJECTIVE: To present a novel, non-invasive echocardiographic application to assess the structural and functional properties of the complex composition of the proximal aorta in patients with end stage renal disease (ESRD). METHODS: 71 haemodialysis patients (mean (SD) age 61.3 (9.3) years, dialysis duration 79.2 (51.6) months) and 62 age matched controls were studied. From the suprasternal view, the distance between ascending and descending aorta was measured with two dimensional ultrasound. The aortic flow wave transit time was measured with pulsed wave Doppler. M mode echocardiography, with simultaneous blood pressure estimates, was used to assess the diameters of the aortic annulus and of the ascending aorta. Pulse pressure, pulse wave velocity (PWV), pressure strain elastic modulus, characteristic impedance, and beta index were calculated. RESULTS: Patients had increased pulse pressure (68.0 (7.2) v 51.4 (5.0) mm Hg, p < 0.001), PWV (6.1 (1.1) v 3.9 (0.6) m/s, p < 0.001), characteristic impedance (174 (58) v 111 (31) m/s.cm2, p < 0.001), pressure strain elastic modulus (872 (254) v 541 (140) mm Hg, p < 0.001), and beta index (8.9 (3.4) v 5.5 (1.4), p < 0.001) compared with controls. In patients PWV was correlated with age and time on haemodialysis (r = 0.44, p < 0.001, and r = 0.51, p < 0.001, respectively). CONCLUSION: A novel application of duplex ultrasound of the proximal aorta showed that patients with ESRD have impaired proximal aortic function compared with controls. The data indicate that these non-invasive measurements can be used to describe status and change in aortic biophysical properties and may be used as a marker for cardiovascular disease risk. PMID- 15814595 TI - Five year outcome after primary coronary intervention for acute ST elevation myocardial infarction: results from a single centre experience. AB - OBJECTIVES: To analyse the five year outcome of unselected patients with acute myocardial infarction (AMI) treated by primary percutaneous coronary intervention (PCI). SETTING: High volume PCI tertiary centre. DESIGN AND RESULTS: The study was based on a sample of 1009 consecutive patients with ST elevation AMI treated by primary PCI. The mean (SD) clinical follow up was 51 (21) months and the follow up rate was 97.8%. The overall mortality was 20% and cardiac mortality was 16%. Non-fatal reinfarction rate was 5% and additional revascularisation procedure rate was 19%. Hospitalisation for heart failure was needed by 42 patients (4%). The variables related to mortality in multivariate Cox analysis were age (hazard ratio (HR) 1.054, 95% confidence interval (CI) 1.039 to 1.069, p < 0.0001), cardiogenic shock (HR 2.985, 95% CI 2.157 to 4.129, p < 0.0001), previous myocardial infarction (HR 1.696, 95% CI 1.199 to 2.398, p = 0.0003), and the presence of multivessel coronary artery disease (HR 1.820, 95% CI 1.317 to 2.514, p = 0.0003). Each additional high risk feature was associated with a relative risk for five year death of 2.328 (95% CI 2.048 to 2.646, p < 0.0001). CONCLUSIONS: The satisfactory results of routine mechanical revascularisation strategy in AMI were maintained during several years of follow up. Patients at risk of death during long term follow up may be identified by simple clinical and angiographic characteristics, such as old age, cardiogenic shock, previous myocardial infarction, and multivessel coronary artery disease. The risk of death progressively increases with the number of these high risk features. PMID- 15814596 TI - Survival trends among Danish patients undergoing coronary angiography for known or suspected ischaemic heart disease: a population based follow up study, 1992 2000. AB - OBJECTIVE: To determine, from population based clinical data, changes in the survival of Danish patients examined by coronary angiography for known or suspected ischaemic heart disease (IHD) during the 1990s. DESIGN: Follow up study. SETTING: The departments of cardiology at Rigshospitalet, Copenhagen University Hospital, and Skejby Hospital, Aarhus University Hospital, Denmark. PATIENTS: Patients with IHD (n = 7021) who underwent first time coronary angiography in 1992, 1996, or 2000. MAIN OUTCOMES MEASURES: Three year survival was compared between cohorts and with that of the general population. Cox proportional hazards regression was used to estimate mortality ratios adjusted for differences in patient characteristics. RESULTS: Survival improved substantially--for example, adjusted mortality ratio was 0.69 (95% confidence interval (CI) 0.55 to 0.87) when comparing patients from 2000 with patients from 1992. The absolute standardised survival rates after three years of follow up were 87.1% (95% CI 85.4% to 88.8%), 89.9% (95% CI 88.5% to 91.3%), and 91.2% (95% CI 90.3% to 92.1%) among patients examined in 1992, 1996, and 2000, respectively. The improvement was not explained by the improvement in overall survival in the general population during the study period. CONCLUSIONS: The survival of Danish patients with known or suspected IHD appears to have improved substantially during the 1990s. PMID- 15814597 TI - Ventricular arrhythmia, Cheyne-Stokes respiration, and death: observations from patients with defibrillators. AB - OBJECTIVE: To determine whether ventricular arrhythmia related to nocturnal hypoxaemia during Cheyne-Stokes respiration (CSR) explains the observation that CSR is an independent marker of death in heart failure. DESIGN: Prospective, observational study. PATIENTS: 101 patients at high risk of clinical serious ventricular arrhythmia fitted with an implantable cardioverter-defibrillator (ICD). MEASUREMENTS: Patients were studied at baseline for CSR during sleep. Arrhythmia requiring device discharge was used as a surrogate marker for possible sudden cardiac death. RESULTS: 101 patients (42 with CSR) were followed up for a total of 620 months. Twenty six patients experienced 432 ICD discharge episodes. Twenty four (6%), 210 (49%), 125 (29%), and 73 (17%) episodes occurred across the time quartiles 0000-0559, 0600-1159, 1200-1759, and 1800-2359, respectively. Kaplan-Meier analysis showed a relative risk of 1 (95% confidence interval 0.5 to 2.2, p = 1) for device discharge in the CSR group. The average (SED) numbers of nocturnal ICD discharges per patient per month of follow up were 0.01 (0.01) and 0.04 (0.02) for patients with and without CSR, respectively (p = 0.6). CONCLUSION: These findings refute the proposition that CSR is related to heart failure death through nocturnal serious ventricular arrhythmia. PMID- 15814598 TI - Antibacterial activity of linezolid and vancomycin in an in vitro pharmacodynamic model of gram-positive catheter-related bacteraemia. AB - OBJECTIVES: The aim of this study was to compare the activity of linezolid and vancomycin in an in vitro pharmacodynamic model to assess potential differences in activity against biofilm-embedded organisms. METHODS: Single-lumen central venous catheters colonized with biofilm-embedded Staphylococcus aureus, Staphylococcus epidermidis or vancomycin-resistant Enterococcus faecium (VRE) were treated with simulated clinical dosing regimens of linezolid 600 mg every 12 h or vancomycin 1 g every 12 h in a one-compartment in vitro pharmacodynamic model. Quantitative cultures were sampled through the catheter and peripheral ports over 48 h to dynamically assess changes in the burden of catheter colonization and organism seeding, respectively. At 24 and 48 h catheters were removed, sonicated and cultured for adherent organisms. RESULTS: Both linezolid and vancomycin suppressed bacterial growth on the catheter and release of S. aureus and S. epidermidis into the model compared with controls (P < 0.05), while linezolid also suppressed counts compared with control and vancomycin versus VRE. Neither agent completely eradicated bacterial colonization of the catheters. MICs for the isolates recovered from the model did not increase over time with linezolid or vancomycin exposure. CONCLUSIONS: Lack of activity against biofilm embedded organisms appeared to be the primary reason for microbiological failure of both drugs in the model. PMID- 15814599 TI - Genome-wide expression profiling of the response to ciclopirox olamine in Candida albicans. AB - OBJECTIVES: The aim of this study was to identify changes in the gene expression profile of Candida albicans upon exposure to the hydroxypyridone anti-infective agent ciclopirox olamine in an effort to better understand its mechanism of action. METHODS: C. albicans SC5314 was exposed to either medium alone or ciclopirox olamine at a concentration equivalent to the IC50 (0.24 mg/L) for 3 h. RNA was isolated and gene expression profiles were compared using DNA microarrays. Differential expression of select genes was confirmed by real-time reverse transcription (RT)-PCR. Mutants disrupted for CDR2 and both CDR1 and CDR2, as well as a clinical isolate overexpressing CDR1 and CDR2, were examined for changes in susceptibility to ciclopirox olamine. RESULTS: A total of 49 genes were found to be responsive to ciclopirox olamine, including 36 up-regulated genes and 13 down-regulated genes. These included genes involved in small molecule transport (HGT11, HXT5, ENA22, PHO84, CDR4), iron uptake (FRE30, FET34, FTR1, FTR2, SIT1) and cell stress (SOD1, SOD22, CDR1, DDR48). Mutants disrupted for CDR2 and both CDR1 and CDR2, as well as a clinical isolate overexpressing CDR1 and CDR2, showed no change in susceptibility to ciclopirox olamine compared with the respective parent. CONCLUSIONS: Consistent with the hypothesis that ciclopirox olamine acts as an iron chelator, it induced changes in expression of many genes involved in iron uptake. Despite induction of the multidrug efflux pump genes CDR1 and, to a lesser extent, CDR2 by ciclopirox olamine, these genes do not affect susceptibility to this agent. PMID- 15814600 TI - Plasmid-borne florfenicol resistance in Pasteurella multocida. AB - OBJECTIVES: A florfenicol-resistant Pasteurella multocida isolate from a calf was investigated for the genetic basis of florfenicol resistance and the location of the resistance gene. METHODS: The P. multocida isolate 381 was investigated for its in vitro susceptibility to antimicrobial agents and its plasmid content. A 10.8 kb florfenicol-chloramphenicol resistance plasmid, designated pCCK381, was identified by transformation into Escherichia coli. The plasmid was mapped with restriction endonucleases, cloned and sequenced completely. RESULTS: Of the antimicrobials tested, plasmid pCCK381 conferred resistance only to chloramphenicol and florfenicol. It showed extended similarity to the 5.1 kb plasmid pDN1 from Dichelobacter nodosus in the part carrying the mobilization and replication genes. An adjacent 3.2 kb segment was highly homologous to the florfenicol resistance gene region of plasmid pMBSF1 from E. coli. In pCCK381, combined resistance to chloramphenicol and florfenicol was based on the presence of a floR gene that showed 97.2-99.7% identity to so far known floR genes. CONCLUSIONS: The results of this study showed that a plasmid-borne floR gene was responsible for chloramphenicol and florfenicol resistance in the bovine respiratory tract pathogen P. multocida. This is, to the best of our knowledge, the first report of a florfenicol resistance gene in a target bacterium. PMID- 15814601 TI - High prevalence of Plasmodium falciparum pfcrt K76T mutation in pregnant women taking chloroquine prophylaxis in Senegal. AB - OBJECTIVES: The risk of malaria infection is increased during pregnancy, and many countries recommend chloroquine prophylaxis in pregnant women, despite Plasmodium falciparum chloroquine resistance. Chloroquine resistance is associated with the pfcrt gene K76T mutation. The aim of this study was to compare the prevalence rate of pfcrt T76 mutation in P. falciparum isolates from infected pregnant and non-pregnant individuals from Senegal. METHODS: The study was conducted in the rural maternity hospital of Thiadiaye, Senegal, where malaria is seasonal. Sixty nine P. falciparum isolates from infected women were collected at delivery. These women were part of a cohort study; they were followed from their first antenatal visit and advised to take chloroquine prophylaxis. For each woman, the earliest P. falciparum-infected blood sample was also used. A control group of 49 non pregnant individuals with asymptomatic P. falciparum infection was enrolled. RESULTS: During pregnancy, prevalence of T76 mutant parasites was higher than in the 49 non-pregnant controls (P<0.001). Among pregnant women, this rate was highest at delivery (P=0.06), and tended to be higher in women who had taken chloroquine prophylaxis, as assessed in urine samples (P=0.08). CONCLUSIONS: Chloroquine prophylaxis is responsible for increased drug consumption and increased drug pressure that may lead to the selection of drug-resistant parasites. This is the first report showing that P. falciparum-infected pregnant women harbour pfcrt T76 mutant parasites more often than non-pregnant individuals, and that the prevalence of this mutation is higher at term than earlier during pregnancy. PMID- 15814602 TI - Combination of nucleoside analogues in the treatment of chronic hepatitis B virus infection: lesson from experimental models. AB - Owing to the persistence of hepatitis B virus (HBV) and the selection of drug resistant mutants, a new concept of antiviral therapy for chronic hepatitis B relies on the combination of nucleoside analogues. In experimental models of HBV infection, several key points concerning these combinations were addressed. (i) Is it possible to achieve a synergic antiviral effect with polymerase inhibitors? (ii) Is it possible to impact on intracellular viral covalently closed circular DNA? (iii) What is the impact of the cross-resistance patterns of the different nucleoside analogues? (iv) What is the effect of viral load suppression on the restoration of specific antiviral cellular responses? The clinical impact of these key issues is discussed in the perspective of new clinical trials. PMID- 15814603 TI - An additional measure for quantifying antibiotic use in hospitals. AB - OBJECTIVES: The number of defined daily doses (DDD) per 100 patient days is often used as an indicator for the selection pressure exerted by antibiotics in the hospital setting. However, this unit of measurement does not fully describe the selection pressure and is sensitive to changes in hospital resource indicators. Additional information is required to facilitate interpretation of this indicator. The number of DDD per 100 admissions could be a valuable additional tool. The aim of this study is to investigate the importance of units of measurement in quantifying antibiotic use data with regards to antibiotic resistance risks. PATIENTS AND METHODS: Trends in antibiotic use in acute care Dutch hospitals between 1997-2001 were studied. Antibiotic use was expressed in DDD per 100 patient days and in DDD per 100 admissions. RESULTS: From 1997 to 2001, total systemic antibiotic use significantly increased from 47.2 to 54.7 DDD per 100 patient days, whereas expressed in DDD per 100 admissions it remained constant. Some individual antibiotics increases in DDD per 100 patient days were not accompanied by increases in DDD per 100 admissions and vice versa. The mean number of total DDD per hospital decreased (not significantly) between 1997 and 2001. The mean number of patient days, admissions and length of stay decreased significantly. CONCLUSIONS: Knowledge of variation in resource indicators and additional expression of the data in DDD per 100 admissions is imperative for a meaningful understanding of observed trends in antibiotic use expressed in DDD per 100 patient days. Further research is needed to determine the correlation between different measures of antibiotic use and the level of antibiotic resistance. PMID- 15814604 TI - TolC but not AcrB is essential for multidrug-resistant Salmonella enterica serotype Typhimurium colonization of chicks. AB - OBJECTIVES: To study the role of the multidrug efflux system AcrAB-TolC in resistance of multidrug-resistant Salmonella enterica serotype Typhimurium (S. Typhimurium) phage type DT104 and DT204 strains to detergents and bile salts. To evaluate the importance of the inner membrane transporter AcrB and the outer membrane component TolC of this efflux system in the colonization of two multidrug-resistant S. Typhimurium DT104 and one DT204 strain in chicks. METHODS: acrB and tolC mutants of multidrug-resistant S. Typhimurium DT104 and DT204 strains were constructed by insertional inactivation of the acrB gene and deletion of the tolC gene. MICs of detergent and bile salts were determined for the wild-type strains, the acrB and the tolC mutant strains, in presence and in absence of the efflux pump inhibitor Phe-Arg beta-naphthylamide. The effect of sodium choleate on the in vitro growth of these strains was also evaluated. The LD50s of the strains were measured in a day-old chicken model, inoculated with several doses (1 x 10(3) to 1 x 10(8) cfu) by the oral route, for 7 days post inoculation. The colonization levels were assessed at the sublethal dose 7 days post-inoculation by determining the number of cfu of Salmonella in the faeces, caecum, spleen and liver. RESULTS: The decrease in resistance levels to bile salts was 64- to 256-fold higher for the tolC mutants than for the acrB mutants relative to those of the parental strains. Addition of choleate in culture medium did not affect the growth of the wild-type strains or that of the acrB mutants, but inhibited completely the growth of the tolC mutants. The LD50s were 1.0 x 10(6) and 1.2 x 10(7) cfu for one wild-type S. Typhimurium DT104 strain and the acrB mutant, respectively, and were >1.0 x 10(8) for the tolC mutants or the S. Typhimurium DT204 strains. In contrast to the acrB mutants, the tolC mutants were unable to colonize the caecum, spleen and liver after 1 week of infection. Moreover, in most chicks, no intestinal excretion was detected for the tolC mutants. The colonization levels of the acrB mutants were not significantly different from those of the wild-type strains. CONCLUSIONS: TolC but not AcrB appears to be essential for multidrug-resistant S. Typhimurium DT104 and DT204 colonization of chicks, which is in accordance with their respective roles in resistance to detergents and bile salts. Therefore, TolC could be a better target than AcrB for the development of efflux pump inhibitors. PMID- 15814605 TI - Induction of interleukin-1beta, tumour necrosis factor-alpha and apoptosis in mouse organs by amphotericin B is neutralized by conjugation with arabinogalactan. AB - OBJECTIVES: To investigate the possibilities that: (i) organ toxicity of amphotericin B-deoxycholate (AMB-DOC) is related to induction of interleukin 1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha) and apoptosis in target organs; and (ii) the reduced toxicity resulting from the conjugation of AMB with water-soluble arabinogalactan (AMB-AG), is related to modulation of these parameters. METHODS: Organ expression of IL-1beta and TNF-alpha was evaluated by enzyme-linked immunosorbent assay (ELISA) in mouse organ biological fluids and in situ by immunohistochemistry. Tissue damage was evaluated histologically, and apoptosis was demonstrated by terminal dUTP nick end labelling (TUNEL) staining. AMB-AG conjugate was compared with the micellar (AMB DOC) and liposomal (AmBisome) AMB formulations. RESULTS: Treatment with AMB-AG or AmBisome caused no observable histopathological damage in the kidneys. In contrast, treatment with AMB-DOC resulted in disruptive changes and apoptosis in renal tubular cells. These effects were found to correlate with induction of high levels of IL-1beta and TNF-alpha in kidney lysates. Unlike AMB-AG, AMB-DOC also induced enhanced IL-1beta and TNF-alpha expression in lysates of lungs, brain, liver and spleen. The marked elevation of these inflammation-apoptosis-promoting cytokines after treatment with AMB-DOC may mediate its systemic and local renal damage. Treatment with AMB-AG (but not AmBisome) appears to uniquely modulate the in situ expression of IL-1beta and enhance secretion of TNF-alpha in kidneys, effects possibly involved in prevention of apoptosis. CONCLUSIONS: AMB-related toxicity is associated with induction of IL-1beta, TNF-alpha and apoptosis in organs. These effects were not observed with AMB-AG conjugate, suggesting its potential as a safer formulation for therapy. PMID- 15814606 TI - Analysis of rpoB and pncA mutations in the published literature: an insight into the role of oxidative stress in Mycobacterium tuberculosis evolution? AB - INTRODUCTION: It is perceived wisdom that within the host macrophage, Mycobacterium tuberculosis frequently encounters oxidative stress. Exposure of bacteria to reactive oxygen intermediates can have a mutagenic effect on the DNA. Various mutations are thought to arise as a consequence, including the oxidation of guanine residues, leading to G?C-->T?A substitution, and oxidation of cytosine resulting in a G?C-->A?T substitution. METHODS: We measured the relative contribution of oxidative stress by recording the percentage of single nucleotide substitutions reported in the genes rpoB and pncA that confer resistance to the antimicrobials rifampicin and pyrazinamide, respectively, and determined whether there is an excess of G?C-->T?A or G?C-->A?T substitutions. RESULTS: Out of 840 clinical isolates reported with single nucleotide mutations in the rpoB gene, 67% were G?C-->A?T changes, and 3% were G?C-->T?A substitutions. These figures were compared to the pncA gene, where out of 114 isolates, 30% of the single nucleotide mutations were G?C-->A?T transitions and 9% were G?C-->T?A changes. CONCLUSIONS: While there is an excess of G?C-->A?T changes in the rpoB gene, this was not the case in the pncA gene. Fifty-three percent of mutations within the rpoB gene were C-->T mutations of the type S531L. Although this mutation gives a fitness disadvantage, it is less than other common mutations, so it is more likely that that fitness is the determinant of surviving mutation rather than oxidative stress because of the small numbers of other C-->T and G-->A mutations at other sites (12%). There was no evidence of oxygen free radicals damaging the guanine bases in either gene. PMID- 15814607 TI - Constitutive UCP3 overexpression at physiological levels increases mouse skeletal muscle capacity for fatty acid transport and oxidation. AB - Uncoupling protein 3 (UCP3) expression is directly correlated to fatty acid oxidation in skeletal muscle. UCP3 has been hypothesized to facilitate high rates of fatty acid oxidation, but evidence thus far is lacking. Our aim was to investigate the effects of UCP3 overexpression and ablation on fatty acid uptake and metabolism in muscle of mice having congenic backgrounds. In mice constitutively expressing the UCP3 protein (human form) at levels just over twofold higher than normal (230% of wild-type levels), indirect calorimetry demonstrated no differences in total energy expenditure (VO2), but a shift toward increased fat oxidation compared with wild-type (WT) mice. Metabolic efficiency (gram weight gain/kcal ingested) was similar between Ucp3 overexpressors, WT and Ucp3 (-/-) mice. In muscle of Ucp3-tg mice, plasma membrane fatty acid binding protein (FABPpm) content was increased compared with WT mice. Although hormone sensitive lipase activity was unchanged across the genotypes, there were increases in carnitine palmitoyltransferase I, beta-hydroxyacylCoA dehydrogenase, and citrate synthase activities and decreases in intramuscular triacylglycerol in muscle of Ucp3-tg mice. There were no differences in muscle mitochondrial content. High-energy phosphates and total muscle carnitine and CoA were also greater in Ucp3-tg compared with WT mice. Taken together, the findings demonstrate an increased capacity for fat oxidation in the absence of significant increases in thermogenesis in Ucp3-tg mice. Findings from Ucp3 (-/-) mice revealed few differences compared with WT mice, consistent with the possibility of compensatory mechanisms. In conjunction with our observed increases in CoA and carnitine in muscle of Ucp3 overexpressors, the findings support the hypothesized role for Ucp3 in facilitating fatty acid oxidation in muscle. PMID- 15814608 TI - Regulation of metabolic transcriptional co-activators and transcription factors with acute exercise. AB - Endurance exercise improves insulin sensitivity and increases fat oxidation, which are partly facilitated by the induction of metabolic transcription factors. Next to exercise, increased levels of FFA's also increase the gene expression of transcription factors, hence making it difficult to discern the effects from contractile signals produced during exercise, from those produced by increased circulatory FFA's. We aimed to investigate, in human skeletal muscle, whether acute exercise affects gene expression of metabolic transcriptional co-activators and transcription factors, including PGC-1alpha, PRC, PPARalpha, beta/delta, and gamma and RXR, SREBP-1c and FKHR, and to discern the effect of exercise per se from those of elevated levels of FFA. Two hours of endurance exercise was performed either in the fasted state, or following carbohydrate ingestion prior to and during exercise, thereby blunting the fasting-induced increase in FA availability and oxidation. Of the genes measured, PGC-1alpha and PRC mRNA increased immediately after, while PPARbeta/delta and FKHR mRNA increased 1-4 h after exercise, irrespective of the increases in FFA's. Our results suggest that the induction in vivo of metabolic transcription factors implicated in mitochondrial biogenesis are under the control of inherent signals, (PGC-1alpha, PRC), while those implicated in substrate selection are under the control of associated signals (PPARbeta/delta, FKHR) stimulated from the contracting skeletal muscle that are independent of circulating FFA levels. PMID- 15814609 TI - Endothelial-like cells expanded from CD34+ blood cells improve left ventricular function after experimental myocardial infarction. AB - Mobilization and recruitment of endothelial progenitor cells (EPC) contributes to vasculogenesis in vivo. So far, applications for cell therapy are limited by the number of available cells. Expansion of EPC or their progeny may, therefore, facilitate its therapeutic use in ischemic disease. The aim of this study was to expand CD34+ EPC-derived progeny from different sources, characterize them, and investigate their potential for use in therapeutic vasculogenesis. CD34+ cells from G-CSF-mobilized peripheral blood (PB) and cord blood (CB) were isolated using immunomagnetic beads and cultured in endothelial cell medium. Cells were expanded up to 16 (PB) and up to 46 (CB) population doublings, respectively. Immunophenotypic and mRNA expression analyses showed a high degree of similarity between the cultured cells and human umbilical vein endothelial cells (HUVEC). By day 14 after transplantation, transplanted human CD31-positive EPC-derived cells were detected. These cells expressed the proliferation marker Ki67 and formed vessel-like structures in ischemic myocardium. Most strikingly, transplantation of EPC-derived cells improved left ventricular function after experimental ischemia, as shown by echocardiography. In conclusion, cells cultured from CD34+ EPC can be expanded in vitro to clinically relevant numbers. In vivo, these cells proliferate, form vascular structures, and improve left ventricular function after experimental myocardial infarction. Therefore, in vitro expanded EPC derived endothelial cells may be beneficial in the treatment of ischemic disease. PMID- 15814610 TI - Differential activities of plant polyphenols on the binding and internalization of cholera toxin in vero cells. AB - Plant polyphenols, RG-tannin, and applephenon had been reported to inhibit cholera toxin (CT) ADP-ribosyltransferase activity and CT-induced fluid accumulation in mouse ileal loops. A high molecular weight fraction of hop bract extract (HBT) also inhibited CT ADP-ribosyltransferase activity. We report here the effect of those polyphenols on the binding and entry of CT into Vero cells. Binding of CT to Vero cells or to ganglioside GM1, a CT receptor, was inhibited in a concentration-dependent manner by HBT and applephenon but not RG-tannin. These observations were confirmed by fluorescence microscopy using Cy3-labeled CT. Following toxin binding to cells, applephenon, HBT, and RG-tannin suppressed its internalization. HBT or applephenon precipitated CT, CTA, and CTB from solution, creating aggregates larger than 250 kDa. In contrast, RG-tannin precipitated CT poorly; it formed complexes with CT, CTA, or CTB, which were demonstrated with sucrose density gradient centrifugation and molecular weight exclusion filters. In agreement, CTA blocked the inhibition of CT internalization by RG-tannin. These data suggest that some plant polyphenols, similar to applephenon and HBT, bind CT, forming large aggregates in solution or, perhaps, on the cell surface and thereby suppress CT binding and internalization. In contrast, RG-tannin binding to CT did not interfere with its binding to Vero cells or GM1, but it did inhibit internalization. PMID- 15814611 TI - Catalysis of thiol/disulfide exchange. Glutaredoxin 1 and protein-disulfide isomerase use different mechanisms to enhance oxidase and reductase activities. AB - Glutaredoxin (Grx) and protein-disulfide isomerase (PDI) are members of the thioredoxin superfamily of thiol/disulfide exchange catalysts. Thermodynamically, rat PDI is a 600-fold better oxidizing agent than Grx1 from Escherichia coli. Despite that, Grx1 is a surprisingly good protein oxidase. It catalyzes protein disulfide formation in a redox buffer with an initial velocity that is 30-fold faster than PDI. Catalysis of protein and peptide oxidation by the individual catalytic domains of PDI and by a Grx1-PDI chimera show that differences in active site chemistry are fundamental to their oxidase activity. Mutations in the active site cysteines reveal that Grx1 needs only one cysteine to catalyze rapid substrate oxidation, whereas PDI requires both cysteines. Grx1 is a good oxidase because of the high reactivity of a Grx1-glutathione mixed disulfide, and PDI is a good oxidase because of the high reactivity of the disulfide between the two active site cysteines. As a protein disulfide reductase, Grx1 is also superior to PDI. It catalyzes the reduction of nonnative disulfides in scrambled ribonuclease and protein-glutathione mixed disulfides 30-180 times faster than PDI. A multidomain structure is necessary for PDI to catalyze effective protein reduction; however, placing Grx1 into the PDI multidomain structure does not enhance its already high reductase activity. Grx1 and PDI have both found mechanisms to enhance active site reactivity toward proteins, particularly in the kinetically difficult direction: Grx1 by providing a reactive glutathione mixed disulfide to supplement its oxidase activity and PDI by utilizing its multidomain structure to supplement its reductase activity. PMID- 15814612 TI - Differential localization of alternatively spliced hypoxanthine-xanthine-guanine phosphoribosyltransferase isoforms in Toxoplasma gondii. AB - A unique feature of the Toxoplasma gondii purine salvage pathway is the expression of two isoforms of the hypoxanthine-xanthine-guanine phosophoribosyltransferase (HXGPRT) of the parasite encoded by a single genetic locus. These isoforms differ in the presence or absence of a 49-amino acid insertion (which is specified by a single differentially spliced exon) but exhibit similar substrate specificity, kinetic characteristics, and temporal expression patterns. To examine possible functional differences between the two HXGPRT isoforms, fluorescent protein fusions were expressed in parasites lacking the endogenous hxgprt gene. Immunoblot analysis of fractionated cell extracts and fluorescence microscopy indicated that HXGPRT-I (which lacks the 49-amino acid insertion) is found in the cytosol, whereas HXGPRT-II (which contains the insertion) localizes to the inner membrane complex (IMC) of the parasite. Simultaneous expression of both isoforms resulted in the formation of hetero oligomers, which distributed between the cytosol and IMC. Chimeric constructs expressing N-terminal peptides from either isoform I (11 amino acids) or isoform II (60 amino acids) fused to a chloramphenicol acetyl transferase (CAT) reporter demonstrated that the N-terminal domain of isoform II is both necessary and sufficient for membrane association. Metabolic labeling experiments with transgenic parasites showed that isoform II or an isoform II-CAT fusion protein (but not isoform I or isoform I-CAT) incorporate [(3)H]palmitate. Mutation of three adjacent cysteine residues within the isoform II-targeting domain to serines blocked both palmitate incorporation and IMC attachment without affecting enzyme activity, demonstrating that acylation of N-terminal isoform II cysteine residues is responsible for the association of HXGPRT-II with the IMC. PMID- 15814613 TI - Suppression of calcium release from inositol 1,4,5-trisphosphate-sensitive stores mediates the anti-apoptotic function of nuclear factor-kappaB. AB - The activation of the transcription factor nuclear factor-kappaB (NF-kappaB) by growth factors, cytokines, and cellular stress can prevent apoptosis, but the underlying mechanism is unknown. Here we provide evidence for an action of NF kappaB on calcium signaling that accounts for its anti-apoptotic function. Embryonic fibroblasts lacking the transactivating subunit of NF-kappaB RelA (p65) exhibit enhanced inositol 1,4,5-trisphosphate (IP(3)) receptor-mediated calcium release and increased sensitivity to apoptosis, which are restored upon re expression of RelA. The size of the endoplasmic reticulum (ER) calcium pool and the number of IP(3) receptors per cell are decreased in response to stimuli that activate NF-kappaB and are increased when NF-kappaB activity is suppressed. The selective antagonism of IP(3) receptors blocks apoptosis in RelA-deficient cells, whereas activation of NF-kappaB in normal cells leads to decreased levels of the type 1 IP(3) receptor and decreased calcium release. Overexpression of Bcl-2 normalizes ER calcium homeostasis and prevents calcium-mediated apoptosis in RelA deficient cells. These findings establish an ER calcium channel as a pivotal target for NF-kappaB-mediated cell survival signaling. PMID- 15814614 TI - Biochemical clustering of monomeric GTPases of the Ras superfamily. AB - To date phylogeny has been used to compare entire families of proteins based on their nucleotide or amino acid sequence. Here we developed a novel analytical platform allowing a systematic comparison of protein families based on their biochemical properties. This approach was validated on the Rho subfamily of GTPases. We used two high throughput methods, referred to as AlphaScreen and FlashPlate, to measure nucleotide binding capacity, exchange, and hydrolysis activities of small monomeric GTPases. These two technologies have the characteristics to be very sensitive and to allow homogenous and high throughput assays. To analyze and integrate the data obtained, we developed an algorithm that allows the classification of GTPases according to their enzymatic activities. Integration and hierarchical clustering of these results revealed unexpected features of the small Rho GTPases when compared with primary sequence based trees. Hence we propose a novel phylobiochemical classification of the Ras superfamily of GTPases. PMID- 15814615 TI - Structure theorems and the dynamics of nitrogen catabolite repression in yeast. AB - By using current biological understanding, a conceptually simple, but mathematically complex, model is proposed for the dynamics of the gene circuit responsible for regulating nitrogen catabolite repression (NCR) in yeast. A variety of mathematical "structure" theorems are described that allow one to determine the asymptotic dynamics of complicated systems under very weak hypotheses. It is shown that these theorems apply to several subcircuits of the full NCR circuit, most importantly to the URE2-GLN3 subcircuit that is independent of the other constituents but governs the switching behavior of the full NCR circuit under changes in nitrogen source. Under hypotheses that are fully consistent with biological data, it is proven that the dynamics of this subcircuit is simple periodic behavior in synchrony with the cell cycle. Although the current mathematical structure theorems do not apply to the full NCR circuit, extensive simulations suggest that the dynamics is constrained in much the same way as that of the URE2-GLN3 subcircuit. This finding leads to the proposal that mathematicians study genetic circuits to find new geometries for which structure theorems may exist. PMID- 15814616 TI - Comparative study of generalized Born models: protein dynamics. AB - In this work, we compare the results of molecular dynamics simulations involving the application of three generalized Born (GB) models to 10 different proteins. The three GB models, the Still, HCT, and modified analytical generalized Born models, were implemented in the computationally efficient gromacs package. The performance of each model was assessed from the backbone rms deviation from the native structure, the number of native hydrogen bonds retained in the simulation, and the experimental and calculated radius of gyration. Analysis of variance (ANOVA) was used to analyze the results of the simulations. The rms deviation measure was found to be unable to distinguish the quality of the results obtained with the three different GB models, whereas the number of native hydrogen bonds and radius of gyration yielded a statistically meaningful discrimination among models. Our results suggest that, of the three, modified analytical generalized Born yields the best agreement between calculated and experimentally derived structures. More generally, our study highlights the need both to evaluate the effects of different variables on the results of simulations and to verify that the results of molecular dynamics simulations are statistically meaningful. PMID- 15814617 TI - (G-H)*-C and G-(C-H)* radicals derived from the guanine.cytosine base pair cause DNA subunit lesions. AB - The radicals generated by the homolytic cleavage of an X-H bond from the guanine.cytosine (G.C) base pair were studied by using carefully calibrated theoretical methods. The gradient-corrected density functional B3LYP was applied in conjunction with double-zeta plus polarization and diffuse function basis sets. Optimized geometries, energies, and vibrational frequencies were obtained for all of the radicals considered. Structural perturbations along with energy relaxation due to radical formation were investigated. Dissociation energies of the G.C base pair and all of the radicals are predicted and compared with the dissociation energy of neutral G.C. The three lowest-energy base pair radicals all involve removal of an H atom from one of the N atoms in G.C. The lowest energy base pair radical has the hydrogen atom removed from the guanine nitrogen atom used for the sugar phosphate linkage in DNA. This (G-H)(*)-C radical has a dissociation energy (to G-H(*) + C) of 30 kcal/mol, which may be compared with 27 kcal/mol for G.C. All of the radicals that are possible outcomes of direct ionizing radiation or oxidizing species were investigated for the presence of local minima with significant structural changes. Major structural deformations cause strain in the interstrand hydrogen bonding in the DNA double helix. Severe geometry changes were observed when the hydrogen was abstracted from interstrand hydrogen bonding sites, along with sizeable energy changes, indicating the potentially serious consequences to the G.C base pair. PMID- 15814618 TI - Reaction coordinates and rates from transition paths. AB - The molecular mechanism of a reaction in solution is reflected in its transition state ensemble and transition paths. We use a Bayesian formula relating the equilibrium and transition-path ensembles to identify transition states, rank reaction coordinates, and estimate rate coefficients. We also introduce a variational procedure to optimize reaction coordinates. The theory is illustrated with applications to protein folding and the dipole reorientation of an ordered water chain inside a carbon nanotube. To describe the folding of a simple model of a three-helix bundle protein, we variationally optimize the weights of a projection onto the matrix of native and nonnative amino acid contacts. The resulting one-dimensional reaction coordinate captures the folding transition state, with formation and packing of helix 2 and 3 constituting the bottleneck for folding. PMID- 15814619 TI - Increased apoptosis in cervical intraepithelial neoplasia associated with HIV infection: implication of oncogenic human papillomavirus, caspases, and Langerhans cells. AB - PURPOSE: Increasing risk of squamous cervical intraepithelial neoplasia (CIN) exits in HIV-infected women. However, the relatively low incidence of invasive carcinoma in the untreated HIV-infected population suggests an imbalance between cell proliferation and apoptosis. We investigated apoptosis and caspases in cervical samples from this population comparatively to non-HIV-infected and control subjects. EXPERIMENTAL DESIGN: Apoptotic terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method, immunohistochemistry for caspase-2, caspase-3, caspase-8, caspase-9, and other apoptosis markers were done on 12 normal cervical samples and 103 low- and high-grade cervical lesions, containing human papillomavirus(es) from 35 HIV-negative and 33 HIV-positive women before tritherapy advent. RESULTS: (a) The apoptotic index (AI) in epithelial cells did not vary between normal mucosa and condyloma acuminata infected or not with HIV. (b) AI augmented with the CIN severity in HIV-positive and HIV-negative women. (c) AI dramatically increased in oncogenic human papillomavirus-infected CIN of HIV-positive population compared with the CIN of similar grade in HIV-negative one. This was associated with a greater expression of caspase-8, active caspase-9, and active caspase-3 in those samples. Moreover, densities of Langerhans' cells, involved in apoptotic bodies engulfment, were greatly reduced in CIN of HIV-positive women. In samples, these densities were highly inversely correlated with AI (r = -0.88, P < 0.002). CONCLUSIONS: This study provides the first evidence for the strongly enhanced apoptosis levels and caspase expression in CIN of untreated HIV-infected women. We suggest that the reduction in Langerhans' cell number could contribute at least partly to apoptotic cell accumulation. PMID- 15814620 TI - Up-regulation of functional chemokine receptor CCR3 in human renal cell carcinoma. AB - There is increasing evidence that chemokines and chemokine receptors are causally involved in tumorigenesis by facilitating tumor proliferation and metastasis. Little is known about the possible function of chemokine receptors in the development and progression of renal cell carcinoma (RCC). We, therefore, analyzed the expression of chemokine receptors in tumor specimens and adjacent healthy kidney tissues [normal kidney cell (NKC)] from 10 RCC patients. We also characterized the permanent RCC cell line A-498. CCR6, CXCR2, and CXCR3 were consistently expressed by both malignant cells and NKCs. A-498 displayed additional expression of CXCR4. Importantly, the expression of CCR3 was almost absent on NKCs but clearly enhanced in a substantial proportion of RCC specimens. The primary CCR3 ligand, eotaxin-1/CCL11, induced intracellular Ca2+ mobilization, receptor internalization, and proliferation in A-498 cells confirming signaling competence of RCC-associated CCR3. In addition, we screened tumor tissue sections of 219 patients and found that 28% (62 of 219) expressed the CCR3 receptor. The presence of CCR3 in tumor samples seemed to correlate with the grade of malignancy. Previous work has established that eotaxin-1 expression is induced by tumor necrosis factor-alpha, a cytokine known to be present in RCC tissue. Our data, therefore, supports a scenario in which eotaxin-1 as part of tumor-associated inflammation promotes progression and dissemination of CCR3 positive RCC. PMID- 15814621 TI - Differential DNA hypermethylation of critical genes mediates the stage-specific tobacco smoke-induced neoplastic progression of lung cancer. AB - Promoter DNA methylation status of six genes in samples derived from 27 bronchial epithelial cells and matching blood samples from 22 former/current smokers and five nonsmokers as well as 49 primary non-small cell lung cancer samples with corresponding blood controls was determined using methylation-specific PCR (MSP). Lung tumor tissues showed a significantly higher frequency of promoter DNA methylation in p16, MGMT, and DAPK (P < 0.05; Fisher's exact test). p16 promoter DNA methylation in tumors was observed at consistently higher levels when compared with all the other samples analyzed (P = 0.001; Fisher's exact test). ECAD and DAPK exhibited statistically insignificant differences in their levels of DNA methylation among the tumors and bronchial epithelial cells from the smokers. Interestingly, similar levels of methylation were observed in bronchial epithelial cells and corresponding blood from smokers for all four genes (ECAD, p16, MGMT, and DAPK) that showed smoking/lung cancer-associated methylation changes. In summary, our data suggest that targeted DNA methylation silencing of ECAD and DAPK occurs in the early stages and that of p16 and MGMT in the later stages of lung cancer progression. We also provide preliminary evidence that peripheral lymphocytes could potentially be used as a surrogate for bronchial epithelial cells to detect altered DNA methylation in smokers. PMID- 15814622 TI - beta-Catenin functions mainly as an adhesion molecule in patients with squamous cell cancer of the head and neck. AB - BACKGROUND: beta-catenin, depending on subcellular localization, plays a dual role in carcinogenesis: as a signaling factor (in the nucleus) and as an adhesion molecule (in cell membrane). In this study, we sought to determine the role of beta-catenin in head and neck carcinogenesis. METHODS: First, we studied the incidence of mutations of beta-catenin in a cohort of 60 head and neck squamous cell cancers (HNSCC). We subsequently evaluated the protein expression levels of beta-catenin in a cohort of oropharyngeal squamous cell cancer tissue microarray using a novel in situ method of quantitative protein analysis and correlated those with cyclin D1 levels and clinical and pathologic data. RESULTS: The mean follow-up time for survivors was 45 months and for all patients was 35 months. We found no mutations in the cohort of 60 HNSCC. beta-catenin displayed primarily membranous expression pattern. Patients with high tumor-node-metastasis stage were more likely to have high expression of beta-catenin (P = 0.040). Patients with low beta-catenin expression had a local recurrence rate of 79% compared with 29% for patients with high beta-catenin tumors (P = 0.0021). Univariate Cox regression revealed a hazard ratio for low beta-catenin tumors of 3.6 (P = 0.004). Kaplan-Meier analysis showed that patients with low beta-catenin expressing tumors trended toward worse 5-year disease-free survival (P = 0.06). In multivariate analysis, only beta-catenin expression status was an independent prognostic factor (P = 0.044) for local recurrence. Tumors with high beta-catenin had low cyclin D1 and vice versa (P = 0.007). CONCLUSIONS: The absence of activating beta-catenin mutations combined with the inverse correlation between beta-catenin levels with cyclin D1 levels and outcome suggest that beta-catenin mainly functions as an adhesion and not signaling molecule in HNSCC. PMID- 15814623 TI - Dedifferentiation precedes invasion in the progression from Barrett's metaplasia to esophageal adenocarcinoma. AB - PURPOSE: Adenocarcinoma arises in Barrett's esophagus by progression from metaplasia to cancer through grades of dysplasia. Our aim in this exploratory study was to characterize the broad changes in gene expression that underlie this histologic progression to cancer and assess the potential for using these gene expression changes as a marker predictive of malignant progression in Barrett's epithelium. EXPERIMENTAL DESIGN: Microarray analysis was used to obtain individual gene expression profiles from endoscopic biopsies of nine esophageal adenocarcinomas and the Barrett's epithelia from which three of the cancers had arisen. Pooled samples from the Barrett's epithelia of six patients without cancer or dysplasia served as a reference. RESULTS: Barrett's epithelia from which cancer had arisen differed from the reference Barrett's epithelia primarily by underexpression of genes, many of which function in governing cell differentiation. These changes in gene expression were found even in those specimens of Barrett's epithelia from which cancer had arisen that lacked dysplasia. Each cancer differed from the Barrett's epithelium from which it had arisen primarily by an overexpression of genes, many of which were associated with tissue remodeling and invasiveness. Cancers without identifiable Barrett's epithelium differed from cancers that had arisen from a Barrett's epithelium by having an even greater number of these overexpressed genes. CONCLUSIONS: Histologic progression from Barrett's epithelium to cancer is associated with a gradient of increasing changes in gene expression characterized by an early loss of gene function governing differentiation that begins before histologic change; gain in function of genes related to remodeling and invasiveness follows later. This correlation of histologic progression with increasing changes in gene expression suggests that gene expression changes in biopsies taken from Barrett's epithelium potentially could serve as a marker for neoplastic progression that could be used to predict risk for developing cancer. PMID- 15814624 TI - Increased mitochondrial DNA content in saliva associated with head and neck cancer. AB - Alterations of the mitochondrial DNA (mtDNA) have been described in human tumors and in other tissues in association with smoking exposure. We did quantitative PCR of cytochrome c oxidase I (Cox I) and cytochrome c oxidase II (Cox II) genes on oral rinse samples obtained from 94 patients with primary head and neck squamous cell carcinoma (HNSC) and a control group of 656 subjects. Mitochondrial DNA/nuclear DNA in saliva from HNSC patients and controls in relationship to smoking exposure, ethanol intake, and tumor stage were examined. Mean levels of Cox I and Cox II in saliva samples were significantly higher in HNSC patients: Cox I, 0.076 [95% confidence interval (95% CI), 0.06-0.09] and Cox II, 0.055 (95% CI, 0.04-0.07) in comparison with controls Cox I, 0.054 (95% CI, 0.05-0.06), P < 0.0001 and Cox II, 0.046 (95% CI, 0.04-0.05), P = 0.003 (t test). MtDNA levels were elevated in primary tumors when compared with matched, pretreatment saliva and significant correlation was noted (Cox I, r = 0.30, P = 0.005 and Cox II r = 0.33, P = 0.002, respectively, Pearson's correlation). On univariate analysis, smoking, age, HNSC diagnosis, and advanced stage of HNSC were associated with higher level of mtDNA content in saliva. Multivariate analysis showed a significant and independent association of HNSC diagnosis, age, and smoking with increasing mtDNA/nuclear DNA for Cox I and Cox II. mtDNA content alteration is associated with HNSC independently of age and smoking exposure, can be detected in saliva, and may be due to elevation in mtDNA content in primary HNSC. PMID- 15814625 TI - Cleavage of L1 in exosomes and apoptotic membrane vesicles released from ovarian carcinoma cells. AB - PURPOSE: The L1 adhesion molecule (CD171) is overexpressed in human ovarian and endometrial carcinomas and is associated with bad prognosis. Although expressed as a transmembrane molecule, L1 is released from carcinoma cells in a soluble form. Soluble L1 is present in serum and ascites of ovarian carcinoma patients. We investigated the mode of L1 cleavage and the function of soluble L1. EXPERIMENTAL DESIGN: We used ovarian carcinoma cell lines and ascites from ovarian carcinoma patients to analyze soluble L1 and L1 cleavage by Western blot analysis and ELISA. RESULTS: We find that in ovarian carcinoma cells the constitutive cleavage of L1 proceeds in secretory vesicles. We show that apoptotic stimuli like C2-ceramide, staurosporine, UV irradiation, and hypoxic conditions enhance L1-vesicle release resulting in elevated levels of soluble L1. Constitutive cleavage of L1 is mediated by a disintegrin and metalloproteinase 10, but under apoptotic conditions multiple metalloproteinases are involved. L1 cleavage occurs in two types of vesicles with distinct density features: constitutively released vesicles with similarity to exosomes and apoptotic vesicles. Both types of L1-containing vesicles are present in the ascites fluids of ovarian carcinoma patients. Soluble L1 from ascites is a potent inducer of cell migration and can trigger extracellular signal-regulated kinase phosphorylation. CONCLUSIONS: We suggest that tumor-derived vesicles may be an important source for soluble L1 that could regulate tumor cell function in an autocrine/paracrine fashion. PMID- 15814626 TI - The p53 codon 72 proline allele is associated with p53 gene mutations in non small cell lung cancer. AB - The p53 gene plays a critical role in cell cycle control, the initiation of apoptosis, and in DNA repair. An Arg/Pro polymorphism at codon 72 of the p53 gene alters the ability of the p53 protein to induce apoptosis, influences the behavior of mutant p53, decreases DNA repair capacity, and may be linked with an increased risk of lung cancer. To further define the role of the p53 codon 72 polymorphism on DNA repair, lung cancer risk, and mutant p53 function, we examined the effect of this polymorphism on mutation of the p53 gene and patient survival in non-small cell lung cancer (NSCLC). Tumor and nonneoplastic (lung or lymphocyte) samples were collected from 182 patients with NSCLC. p53 mutations were detected by direct sequencing and/or the Gene Chip p53 assay in 93 of 182 (51%) tumors. p53 codon 72 polymorphisms were identified by PCR/RFLP analysis. p53 mutations were significantly (P = 0.01) associated with the number of codon 72 Pro alleles: Pro/Pro homozygotes, 17 of 26 (65%); Arg/Pro heterozygotes, 45 of 79 (57%); and Arg/Arg homozygotes, 31 of 77 (40%). The number of codon 72 Pro alleles was independently associated with p53 mutations (odds ratio, 1.97; 95% confidence interval, 1.14-3.40; P = 0.01) in a multiple logistic regression model. The codon 72 polymorphism did not influence patient survival in either the entire patient group or among patients with p53 mutant tumors. In summary, the p53 Pro allele is associated with an increased frequency of p53 mutations in NSCLC. PMID- 15814627 TI - Mcm2, Geminin, and KI67 define proliferative state and are prognostic markers in renal cell carcinoma. AB - PURPOSE: The origin licensing factors minichromosome maintenance 2 (Mcm2) and Geminin have recently been identified as critical regulators of growth and differentiation. Here we have investigated the regulation of these licensing factors together with Ki67 to further elucidate the cell cycle kinetics of renal cell carcinoma (RCC). Furthermore, we have examined the role of Ki67, Mcm2, and Geminin in disease-free survival after nephrectomy in patients with localized RCC. EXPERIMENTAL DESIGN: Tissue sections from 176 radical nephrectomy specimens were immunohistochemically stained with Mcm2, Geminin, and Ki67 antibodies. Labeling indices (LI) for these markers were compared with clinicopathologic parameters (median follow-up 44 months). RESULTS: In RCC, Mcm2 is expressed at much higher levels than Ki-67 and Geminin, respectively [medians 41.6%, 7.3%, and 3.5% (P < 0.001)] and was most closely linked to tumor grade (P < 0.001). For each marker, Kaplan-Meier survival curves provided strong evidence that increased expression is associated with reduced disease-free survival time (P < 0.001). Additionally, an Mcm2-Ki67 LI identified a unique licensed but nonproliferating population of tumor cells that increased significantly with tumor grade (P = 0.004) and was also of prognostic value (P = 0.01). On multivariate analysis, grade, vascular invasion, capsular invasion, Ki67 LI >12%, and age were found to be independent prognostic markers. CONCLUSIONS: Although Ki67 is identified as an independent prognostic marker, semiquantitative assessment is difficult due to the very low proliferative fraction identified by this marker. In contrast, Mcm2 identifies an increased growth fraction that is closely linked to grade, provides prognostic information, and is amenable to semiquantitative analysis in routine pathologic assessment. PMID- 15814628 TI - Nuclear factor-kappaB activation correlates with better prognosis and Akt activation in human gastric cancer. AB - PURPOSE: Because the biological significance of constitutive nuclear factor kappaB (NF-kappaB) activation in human gastric cancer is unclear, we undertook this study to clarify the regulatory mechanism of NF-kappaB activation and its clinical significance. EXPERIMENTAL DESIGN: Immunohistochemistry for NF kappaB/RelA was done on 290 human gastric carcinoma specimens placed on tissue array slides. The correlations between NF-kappaB activation and clinicopathologic features, prognosis, Akt activation, tumor suppressor gene expression, or Bcl-2 expression were analyzed. We also did luciferase reporter assay, Western blot analysis, and reverse transcription-PCR using the SNU-216 human gastric cancer cell line transduced with retroviral vectors containing constitutively active Akt or the NF-kappaB repressor mutant of IkappaBalpha. RESULTS: Nuclear expression of RelA was found in 18% of the gastric carcinomas and was higher in early-stage pathologic tumor-node-metastasis (P = 0.019). A negative correlation was observed between NF-kappaB activation and lymphatic invasion (P = 0.034) and a positive correlation between NF-kappaB activation and overall survival rate of gastric cancer patients (P = 0.0228). In addition, NF-kappaB activation was positively correlated with pAkt (P = 0.047), p16 (P = 0.004), adenomatous polyposis coli (P < 0.001), Smad4 (P = 0.002), and kangai 1 (P < 0.001) expression. An in vitro study showed that NF-kappaB activity in gastric cancer cells is controlled by and controls Akt. CONCLUSIONS: NF-kappaB activation was frequently observed in early stage gastric carcinoma and was significantly correlated with better prognosis and Akt activation. These findings suggest that NF-kappaB activation is a valuable prognostic variable in gastric carcinoma. PMID- 15814629 TI - The number of lymph node metastases in gastric cancer correlates with the angiotensin I-converting enzyme gene insertion/deletion polymorphism. AB - PURPOSE: In the present study, we aimed to substantiate the putative significance of angiotensin I-converting enzyme (ACE) on gastric cancer biology by investigating the influence of its gene polymorphism on gastric cancer progression. EXPERIMENTAL DESIGN: Genomic DNA was purified from peripheral blood mononuclear cells or tissue specimens. Amplified ACE gene fragments were separated on agarose gels. D or I alleles were identified by the presence of 190- or 490-bp fragments, respectively. Local expression of ACE was investigated by immunohistochemistry. RESULTS: Twenty-four of 113 (21%) gastric cancer patients had the II, 57 (51%) the ID, and 32 (28%) the DD genotype. The distribution of the ACE genotypes did not differ significantly from the control group of 189 patients without gastric cancer. However, the ACE genotypes correlated with the number of lymph node metastases and the Unio Internationale Contra Cancrum (UICC) tumor stage. Patients with the II genotype had a highly significantly smaller number of lymph node metastases (P < 0.001) and a significantly lower UICC tumor stage (P = 0.01) than patients with the DD genotype. No correlation was found between tumor type, tumor location, local tumor growth, distant metastases, and the ACE genotype. The expression of ACE in gastric cancer was investigated by immunohistochemistry in 100 of 113 patients. ACE was expressed by endothelial cells in all (100%) specimens and by tumor cells in 56 (56%) specimens. CONCLUSIONS: Our study shows that ACE is expressed locally in gastric cancer and that the gene polymorphism influences metastatic behavior. PMID- 15814630 TI - Prognostic significance of tissue factor in pancreatic ductal adenocarcinoma. AB - Tissue factor (TF) is a transmembrane glycoprotein that plays roles in the blood coagulation and intracellular signaling pathways, and has also been suggested to modulate the biological behavior of cancer cells. In order to examine the clinicopathologic significance of TF expression in pancreatic ductal adenocarcinoma, TF expression was determined by immunohistochemistry using a newly raised anti-TF monoclonal antibody in 113 patients who had undergone surgical resection of pancreatic ductal adenocarcinoma. According to the incidence of tumor cell immunopositivity, patients were divided into "negative TF" (0%), "weak TF" (<25%), or "high TF" (25% or more) groups, which accounted for 11.6% (n = 13), 44.2% (n = 50), and 44.2% (n = 50) of the total, respectively. Increased TF expression was correlated with the extent of the primary tumor (P = 0.0043), lymph node metastasis (P = 0.0043), lymphatic distant metastasis (P = 0.0039), advanced tumor-node-metastasis stage (P = 0.0002), and high tumor grade (P = 0.0164). Multivariate analysis using the Cox proportional hazards model showed that high TF expression was an independent negative predictor for survival (hazard ratio, 2.014; P = 0.0076). Moreover, patients with TF-negative tumors had a significantly better prognosis even if lymph node metastasis was present (P < 0.0001). We also showed that TF knockdown by RNA interference suppressed the invasiveness of a pancreatic adenocarcinoma cell line in vitro. These results indicate that TF expression may contribute to the aggressiveness of pancreatic ductal adenocarcinoma by stimulating tumor invasiveness, and that evaluation of the primary tumor for TF expression may identify patients with a poor prognosis. PMID- 15814631 TI - Minimal residual disease in acute myeloid leukemia is predicted by an apoptosis resistant protein profile at diagnosis. AB - PURPOSE: Apoptosis is an important mechanism regulating survival of acute myeloid leukemia cells. The apoptosis-related protein profile at diagnosis is important for achieving complete remission thereby affecting survival variables such as disease-free survival (DFS) and overall survival (OS).To investigate the role of the apoptosis protein profile in further response to therapy and outgrowth of disease. EXPERIMENTAL DESIGN: We studied whether Bcl-2, Bcl-xL, Mcl-1, Bax as well as the Bcl-2/Bax ratio and a combination of all (antiapoptosis index, AAI) are related to the frequency of malignant cells surviving the chemotherapy (i.e., minimal residual disease, MRD). MRD cells were identified by leukemia-associated aberrant phenotypes established at diagnosis by flow cytometry. RESULTS: We found that Bcl-2 (R = 0.55, P = 0.002), Bcl-2/Bax (R = 0.42, P = 0.02), and AAI (R = 0.47, P = 0.01) at diagnosis directly correlated with MRD after the first cycle of chemotherapy. In turn, MRD frequency after first cycle correlated with DFS (P = 0.04). Taken together, these results directly explain why Bcl-2/Bax and especially AAI (P = 0.007) at diagnosis correlate with DFS. CONCLUSION: Our results show that apoptosis resistance plays an important role in the first stage of the therapy (i.e., to eliminate the bulk of malignant cells), in terms of achievement of complete remission and frequency of MRD after first cycle of therapy. PMID- 15814632 TI - Molecular detection of early-stage laryngopharyngeal squamous cell carcinomas. AB - PURPOSE: We sought to determine whether early-stage laryngopharyngeal squamous cell carcinomas (SCC) can be detected through molecular analysis of exfoliated cells collected with the use of a pharyngoesophageal brush (PEB). EXPERIMENTAL DESIGN: Thirty-three patients with a single, untreated, early-stage (T1 or T2) SCC of the supraglottic larynx or pharynx underwent collection of cells with a PEB, followed by endoscopic biopsy of the tumor. PEB specimens were also collected from five healthy subjects. PEB samples and tumor tissue were examined for hypermethylation of p16INK4a (CDKN2) gene promoter CpG islands (assayed by methylation-specific PCR) and UT5085 tetranucleotide microsatellite instability (assayed by GeneScan analysis). PEB samples were also subjected to cytologic analysis. RESULTS: Eight of 33 (24%) tumors exhibited a bandshift at UT5085, and 14 of 33 (42%) exhibited hypermethylation at the p16 promoter. Overall, 17 of 33 (52%) patients had at least one of the two markers in their tumor. Cytologic analysis of PEB samples revealed tumor in 4 of 33 (12%) patients; cytologic findings were normal in all five control subjects. Molecular analysis of PEB samples revealed tumor DNA in 13 of 17 (76%) patients with at least one of the two molecular markers in their tumor. Eight of 14 (57%) patients with p16 hypermethylation in their tumor and 8 of 8 (100%) patients with UT5085 microsatellite instability in their tumor had similar findings in the PEB samples. None of the PEB samples from the control subjects or patients with neither molecular marker in their tumor displayed abnormality. CONCLUSION: Molecular analysis of PEB samples holds promise for the early detection of early stage laryngopharyngeal SCCs. New molecular markers need to be identified to increase the sensitivity of molecular screening. PMID- 15814633 TI - Defects in the human leukocyte antigen class I antigen processing machinery in head and neck squamous cell carcinoma: association with clinical outcome. AB - PURPOSE: Human leukocyte antigen (HLA) class I antigen defects, which are frequently present in head and neck squamous cell carcinoma (HNSCC) cells may provide the tumor with an escape mechanism from immune surveillance. Scanty information is available about mechanisms underlying HLA class I antigen defects in both lesions and cell lines from HNSCC. In this study, we investigate the role of antigen processing machinery (APM) component abnormalities in the generation of deficient HLA class I surface expression of HNSCC cells. EXPERIMENTAL DESIGN: Using immunohistochemistry, Western blot, and RT-PCR analyses we correlated the expression of the IFN-gamma inducible proteasome subunits and of the peptide transporter TAP with that of HLA class I antigens in biopsies and cell lines from primary, recurrent, and metastatic HNSCC. Furthermore, APM component and HLA class I antigen expression in surgically removed lesions were correlated with the course of the disease in order to assess the clinical significance of deficient expression of these molecules. RESULTS: A high frequency of LMP2, LMP7, and TAP1 down-regulation or loss was found in tumor lesions and cell lines obtained from HNSCC cancer patients. These defects could be corrected by incubating cells with IFN-gamma. Furthermore, LMP2, LMP7, TAP1, TAP2, and HLA class I antigen expression rates in primary HNSCC lesions were found to predict overall survival. Lastly, the level of LMP7 expression was significantly associated with disease recurrence at 2 years. CONCLUSIONS: Our results suggest that the analysis of APM component expression in HNSCC lesions can provide useful prognostic information in patients with HNSCC. PMID- 15814634 TI - Messenger RNA expression levels of CXCR4 correlate with metastatic behavior and outcome in patients with osteosarcoma. AB - PURPOSE: To determine if osteosarcoma cells express chemokine receptors and if their presence or absence relates to clinical features. EXPERIMENTAL DESIGN: Using fluorescent quantitative real-time PCR, the pattern of 17 chemokine receptors in 3 osteosarcoma cell lines and 68 osteosarcoma patient samples was analyzed. RESULTS: The expression of the chemokine receptors was generally low among the cell lines. In the high-grade osteosarcoma patient samples (n = 47), CXCR4 was the most commonly expressed (63%) and its expression level was inversely correlated to overall survival (P < 0.0001), event-free survival (P < 0.001), and metastasis-free survival (MFS; P = 0.002). There was also a correlation between the expression level of CXCR4 and the presence of metastasis at diagnosis (P = 0.002). CCR7 was expressed in 43% of the samples and its expression level was inversely correlated with overall survival (P = 0.03) and MFS (P = 0.007). CCR10 mRNA expression level was inversely correlated with MFS (P = 0.009). There was no association between the expression of CXCR4, CCR7, and CCR10. Of the 26 samples studied for stromal cell-derived factor-1 expression, 77% expressed it, but there was no correlation with the clinical variables or CXCR4 expression. Multivariate analysis revealed that mRNA expression level of CXCR4 was the only significant variable for overall survival (P = 0.0006), event free survival (P = 0.004), and MFS (P = 0.025). CONCLUSIONS: These data suggest that CXCR4 could be useful as a prognostic factor and as a predictor of potential metastatic development in osteosarcoma. If further studies confirm that it is relevant to metastases in this disease, it could represent a new therapeutic target. PMID- 15814635 TI - Opposite roles of human pancreatitis-associated protein and REG1A expression in hepatocellular carcinoma: association of pancreatitis-associated protein expression with low-stage hepatocellular carcinoma, beta-catenin mutation, and favorable prognosis. AB - PURPOSE: Pancreatitis-associated protein (PAP) and regenerating protein 1 alpha (Reg1A) are up-regulated during the pancreas regeneration. This study is to investigate the clinicopathologic denotation of their expression in hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: PAP and REG1A mRNA levels were measured in 265 surgically removed unifocal primary HCCs using reverse transcription-PCR. RESULTS: PAP and REG1A mRNAs were detected in 97 (36.6%) and 55 (20.8%) HCCs, respectively, including 46 with coexpression but in none of the 219 nontumorous livers. HCCs with PAP expression correlated with low-stage tumors without evidence of vascular invasion (P = 0.013) but the REG1A expression did not. By a combination analysis, HCCs with PAP expression alone showed the lowest frequency of p53 mutation (P < 0.036), the highest rates of grade 1 and low-stage tumors (P < 0.007 and P < 0.001, respectively), less frequent early tumor recurrence (P = 0.051), and hence a better 5-year survival (P = 0.044) than groups expressing PAP and REG1A, REG1A alone, and neither PAP or REG1A. Besides, PAP expressing HCCs had significantly frequent beta-catenin mutation, regardless of REG1A expression, P < 0.00001. In the subset of HCCs that has no mutations of p53 and beta-catenin but showed PAP expression, coexpression of REG1A and PAP was associated with more frequent vascular invasion than PAP expression alone (P < 0.005). CONCLUSIONS: These data suggest that PAP expression designate a subset of low-grade, low-stage HCC with frequent beta-catenin mutation and hence more favorable prognosis, whereas further genetic or epigenetic alterations, such as p53 mutation and REG1A expression, lead to more advanced HCCs. PMID- 15814636 TI - Prognostic value of tumor-infiltrating dendritic cells in colorectal cancer: role of maturation status and intratumoral localization. AB - The clinical significance of tumor-infiltrating dendritic cells has been reported in a variety of human solid tumors as shown by the correlations found between the presence of tumor-infiltrating dendritic cells and clinical prognosis. In this study, we evaluated whether there is an association between the presence and maturation status of tumor-infiltrating dendritic cells, T lymphocytes, and clinical course in 104 primary tumor samples of patients with colorectal cancer. Dendritic cells were identified with four different markers (S-100, HLA class II, CD208, and CD1a) in double immunohistochemistry, with laminin as second marker to support the exact localization. Tumor-infiltrating dendritic cells showed a distinct infiltration pattern based on their maturation status. CD1a-positive dendritic cells resided in the advancing tumor margins in relatively high numbers, whereas mature CD208-positive dendritic cells were sparsely present in the tumor epithelium but mainly distributed in the tumor stroma and advancing tumor margin. Furthermore, high infiltration of CD1a-positive dendritic cells in the tumor epithelium was significantly correlated to the infiltration of CD4 lymphocytes (P = 0.006). Patients with relatively high numbers of mature CD208 positive infiltrating dendritic cells in the tumor epithelium had a shorter overall survival (P = 0.004). In addition, patients with relatively high numbers of CD1a-positive dendritic cells in the advancing margin of the tumor had a shorter disease-free survival (P = 0.03). We found that tumor-infiltrating dendritic cells had preferential infiltration sites within a tumor, affected local tumor cell-immune cell interactions, and correlated to the clinical prognosis of colorectal cancer patients. PMID- 15814637 TI - Expression of hypoxia-inducible factor-1alpha protein predicts survival in patients with transitional cell carcinoma of the upper urinary tract. AB - Hypoxia-inducible factor-1 (HIF-1), identified as one of the transcription factors, has been found to play an essential role in oxygen homeostasis. HIF-1 is a heterodimer composed of HIF-1alpha and HIF-1beta. Increased levels of HIF 1alpha have been reported during the carcinogenesis and progress of several tumors. We investigated the prognostic importance of HIF-1alpha expression in transitional cell carcinoma of the upper urinary tract. In 127 cases of transitional cell carcinoma of the upper urinary tract, we examined its expression (using immunohistochemistry and in situ hybridization), and also its relation to the expression of p53 oncoprotein, as well as to proliferating cell nuclear antigen (PCNA) immunoreactivity, microvessel density, clinicopathologic parameters, and clinical outcome. A positive expression of HIF-1alpha protein was recognized in 55.1% of samples, the expression being apparent within the nucleus in tumor cells. HIF-1alpha protein expression correlated with grade, growth pattern, p53 oncoprotein expression, and PCNA index, but not with stage. Furthermore, a significant correlation was found between HIF-1alpha protein expression and both overall and disease-free survival rates in the univariate and multivariate analyses (in all tumors and in invasive tumors). A positive expression of HIF-1alpha mRNA was recognized in 69.6% of 125 samples which were available, the expression being apparent within the cytoplasm in tumor cells. The positive expression of HIF-1alpha mRNA by in situ hybridization correlated significantly with HIF-1alpha protein expression by immunohistochemistry. HIF 1alpha mRNA expression only correlated with pattern of growth (P = 0.0078). In conclusion, the detection of HIF-1alpha protein would seem to be of value in informing the prognosis of transitional cell carcinoma of the upper urinary tract. PMID- 15814638 TI - Prostate stem cell antigen is overexpressed in prostate cancer metastases. AB - PURPOSE: Prostate stem cell antigen (PSCA) is expressed by a majority of prostate cancers and is a promising therapeutic target. PSCA protein and mRNA expression was examined in prostate cancer bone, lymph node, and visceral metastases to assess the potential of PSCA as an immunotherapeutic target in advanced prostate cancer. EXPERIMENTAL DESIGN: Immunohistochemical analysis of PSCA protein expression and quantitative mRNA expression analysis of PSCA was done on clinical specimens of prostate cancer bone, lymph node, and visceral metastases. PSCA protein and mRNA expression levels were quantified and compared between available matched pairs of bone and lymph node or visceral metastases. RESULTS: Bone metastases stained with higher intensity of PSCA compared with lymph node or liver metastases in seven of eight (87.5%) matched pairs (P = 0.035). PSCA mRNA expression was equal or greater than that of LAPC-9, a PSCA expressing xenograft, in 12 of 24 (50%) cases of prostate cancer metastases and was significantly correlated with PSCA protein expression (sigma = 0.84, P = 0.0019). Overall, PSCA protein expression was detected in 41 of 47 (87.2%), four of six (66.7%), and two of three (66.7%) cases of bone, lymph node, and liver metastases, respectively. Mean PSCA staining intensity was significantly higher in prostate cancer bone metastases compared with lymph node metastases (2.0 +/- 0.02 versus 0.83 +/- 0.31, P = 0.014). CONCLUSIONS: Prostate cancer metastases express PSCA. However, greater PSCA staining intensity and level of PSCA mRNA expression was associated with bone metastases compared with lymph node metastases. This study suggests that PSCA is a promising tumor marker and potential therapeutic target for patients with metastatic prostate cancer. PMID- 15814639 TI - Prognostic significance of fascin overexpression in human esophageal squamous cell carcinoma. AB - PURPOSE: Fascin, an actin bundling protein, induces membrane protrusions and increased cell motility in various transformed cells. The expression of fascin in epithelial neoplasms has been described only recently, and the role of fascin in esophageal squamous cell carcinoma (ESCC) is still unknown. EXPERIMENTAL DESIGN: Paraffin sections of 200 patients with ESCC were immunohistochemically investigated. The expression levels of fascin mRNA in 20 ESCC tissues were compared with that in corresponding normal esophageal epithelium by semiquantitative reverse transcription-PCR. We also examined fascin protein expression in 33 ESCC cell lines. The role of fascin in cell motility and invasiveness in ESCC cells was assessed by the vector-based small interfering RNA. RESULTS: In immunohistochemical study, the intensity of fascin expression was usually increased in the tumor compared with that in normal epithelium. Fascin overexpression was significantly associated with a poor prognosis (immunoreactive rate, P = 0.033; immunoreactive intensity, P = 0.031). The fascin immunoreactive rate was associated with extent of the tumor (P = 0.002) and lymph node metastasis (P = 0.003). Multivariate analysis showed that fascin expression intensity was an independent prognostic factor, but the immunoreactive rate was not. In addition, up-regulation of fascin mRNA was found in 60% (12 of 20) of patients. In vitro study revealed that all 33 ESCC cell lines expressed fascin protein at a certain level. KYSE170, one of the fascin-overexpressed cells, decreased its motile and invasive properties after down-regulation of fascin expression. CONCLUSION: Our findings suggest that fascin overexpression may play an important role in the progression of ESCC. PMID- 15814640 TI - SMAD4 as a prognostic marker in colorectal cancer. AB - More than 50% of patients with Dukes C colorectal cancer have disease recurrence and die within 5 years after surgical removal of their primary tumor. It is currently not possible to distinguish patients with good and bad prognosis. SMAD4 is an important tumor suppressor gene that mediates transforming growth factor beta superfamily signaling and is located in chromosome 18q21, a region with frequent genetic losses in these tumors. Allelic imbalance in 18q has been linked to poor prognosis in a subset of colorectal cancer patients. Therefore, we generated a tissue microarray containing triplicate tumor samples from 86 Dukes C patients and used immunohistochemistry to assess the relative expression level of SMAD4 and its value as a prognostic marker. In addition, SMAD4 was screened for mutations and two polymorphic microsatellite markers were used to assess the presence of allelic imbalance in these tumors. Patients with tumors expressing high SMAD4 levels had significantly better overall (P < 0.025) and disease-free (P < 0.013) survival than patients with low levels. This identifies SMAD4 as a prognostic marker for Dukes C colorectal cancer. Although all tumors with absent SMAD4 staining showed allelic imbalance in 18q21, tumors with 18q21 allelic imbalance as a group showed no difference in SMAD4 levels compared with tumors without allelic imbalance, suggesting that additional mechanisms of SMAD4 down regulation exist. In addition, although SMAD4 mutations were found in five tumors, they were not associated with shorter survival. In conclusion, the level of expression of SMAD4 was found to be a more sensitive marker than 18q21 allelic imbalance and SMAD4 mutations, which were of no prognostic significance for these patients. PMID- 15814641 TI - Variance in the expression of 5-Fluorouracil pathway genes in colorectal cancer. AB - Although colorectal cancer has the third highest cancer mortality rate, the treatment remains far from optimized with patients showing variable responses to standard treatment. Molecular differences in pharmacologically relevant genes may contribute to the variability in response. This study used Taqman PCR to investigate the expression of 24 5-fluorouracil (5-FU) pathway genes in colorectal cancer using paired nontumor and tumor sample from 52 patients with Dukes' C colon cancer. In comparing tumor versus nonmalignant tissue, 14 of the 24 genes showed significant variation in gene expression. For 11 of these same genes (FPGS, DHFR, GGH, NME1, NME2, RRM2, UMPH2, UNG, UMPS, TP53, and TK1), a significant proportion of the patients showed an over expression of the particular gene in tumor tissue with a tumor-to-nonmalignant (T/N) ratio >1.2, whereas one gene (DPYD) showed the converse with a large number of patients showing a lower expression in the tumor tissue (T/N < 0.8). Multiple gene correlations for the genes of the 5-FU pathway were found with the Spearman rank correlation of >0.6 (all P > 0.001), suggesting possible coregulation mechanisms. Hierarchical clustering analysis created at least three groups of genes, which were consistent with groupings by the other statistical methods. Additionally, the hierarchical clustering showed two distinct groups of patients based on their gene expression. These variations in gene expression could provide valuable insights for optimizing treatment selection for patients with colorectal cancer. PMID- 15814642 TI - Severe drug toxicity associated with a single-nucleotide polymorphism of the cytidine deaminase gene in a Japanese cancer patient treated with gemcitabine plus cisplatin. AB - PURPOSE: We investigated single-nucleotide polymorphisms of the cytidine deaminase gene (CDA), which encodes an enzyme that metabolizes gemcitabine, to clarify the relationship between the single-nucleotide polymorphism 208G>A and the pharmacokinetics and toxicity of gemcitabine in cancer patients treated with gemcitabine plus cisplatin. EXPERIMENTAL DESIGN: Six Japanese cancer patients treated with gemcitabine plus cisplatin were examined. Plasma gemcitabine and its metabolite 2',2'-difluorodeoxyuridine were measured using an high-performance liquid chromatography method, and the CDA genotypes were determined with DNA sequencing. RESULTS: One patient, a 45-year-old man with pancreatic carcinoma, showed severe hematologic and nonhematologic toxicities during the first course of chemotherapy with gemcitabine and cisplatin. The area under the concentration time curve value of gemcitabine in this patient (54.54 microg hour/mL) was five times higher than the average value for five other patients (10.88 microg hour/mL) treated with gemcitabine plus cisplatin. The area under the concentration-time curve of 2',2'-difluorodeoxyuridine in this patient (41.58 microg hour/mL) was less than the half of the average value of the five patients (106.13 microg hour/mL). This patient was found to be homozygous for 208A (Thr70) in the CDA gene, whereas the other patients were homozygous for 208G (Ala70). CONCLUSION: Homozygous 208G>A alteration in CDA might have caused the severe drug toxicity experienced by a Japanese cancer patient treated with gemcitabine plus cisplatin. PMID- 15814643 TI - Predicting response to methotrexate, vinblastine, doxorubicin, and cisplatin neoadjuvant chemotherapy for bladder cancers through genome-wide gene expression profiling. AB - PURPOSE: Neoadjuvant chemotherapy for invasive bladder cancer, involving a regimen of methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC), can improve the resectability of larger neoplasms for some patients and offer a better prognosis. However, some suffer severe adverse drug reactions without any effect, and no method yet exists for predicting the response of an individual patient to chemotherapy. Our purpose in this study is to establish a method for predicting response to the M-VAC therapy. EXPERIMENTAL DESIGN: We analyzed gene expression profiles of biopsy materials from 27 invasive bladder cancers using a cDNA microarray consisting of 27,648 genes, after populations of cancer cells had been purified by laser microbeam microdissection. RESULTS: We identified dozens of genes that were expressed differently between nine "responder" and nine "nonresponder" tumors; from that list we selected the 14 "predictive" genes that showed the most significant differences and devised a numerical prediction scoring system that clearly separated the responder group from the nonresponder group. This system accurately predicted the drug responses of 8 of 9 test cases that were reserved from the original 27 cases. Because real-time reverse transcription-PCR data were highly concordant with the cDNA microarray data for those 14 genes, we developed a quantitative reverse transcription-PCR-based prediction system that could be feasible for routine clinical use. CONCLUSIONS: Our results suggest that the sensitivity of an invasive bladder cancer to the M VAC neoadjuvant chemotherapy can be predicted by expression patterns in this set of genes, a step toward achievement of "personalized therapy" for treatment of this disease. PMID- 15814644 TI - Evasion of a single-step, chemotherapy-induced senescence in breast cancer cells: implications for treatment response. AB - PURPOSE: The purpose of this study is to define the mechanistic basis for recovery of proliferative capacity in breast tumor cells after chemotherapy. Here, we test the hypothesis that evasion of senescence confers resistance to chemotherapeutic drugs and ionizing radiation. EXPERIMENTAL DESIGN: MCF-7 cells were treated with a single, clinically relevant dose (0.75-1.0 micromol/L) of Adriamycin. Two weeks following induction of senescence, clonal outgrowths were expanded and characterized in terms of senescence-associated beta-galactosidase activity, gene expression profiles (Affymetrix U95 probe sets, Affymetrix, Santa Clara, CA) with confirmatory Western analyses, and telomerase activity following a second drug treatment. Levels of intracellular Adriamycin, as well as cross resistance to other therapeutic agents, were also determined to define the resistance phenotype. RESULTS: A senescence-resistant (SR) clone (clone 2) was identified that was largely refractory to both Adriamycin-induced and gamma irradiation-induced senescence. Clone 2 continued to proliferate and maintain high levels of telomerase activity following a second drug treatment, when treated parental cells expressed very low levels of telomerase and many positive cell cycle regulators. SR clone 2 also expressed substantially more cdc-2 than parental cells and undetectable levels of MDR1, showed an intact p53 checkpoint and only a modestly lower level of intracellular drug accumulation, while exhibiting cross-resistance to other topoisomerase inhibitors. CONCLUSIONS: SR clone 2 is intrinsically resistant to DNA damage-induced senescence perhaps through an ability to prevent down-regulation of cdc-2. Telomerase is a marker of proliferative recovery for breast cancer cells after chemotherapy exposure. Evasion or escape from a single-step, drug-induced senescence may represent a unique and previously unrecognized drug-resistance phenotype. PMID- 15814646 TI - Anti-flt1 peptide, a vascular endothelial growth factor receptor 1-specific hexapeptide, inhibits tumor growth and metastasis. AB - PURPOSE: The purpose of this study was to develop antagonists specific for the vascular endothelial growth factor receptor 1 (VEGFR1) and to investigate the effects of the antagonists on the VEGF-induced endothelial cell functions and tumor progression. EXPERIMENTAL DESIGN: Hexapeptides that inhibit binding of VEGFR1 and VEGF were identified through screening of synthetic peptide library. A selected peptide, anti-Flt1, was investigated for binding specificity with various receptors and ligand peptides. Effects of the peptide on proliferation, cell migration, and fibrin gel-based angiogenesis of endothelial cells were also investigated. The activity of anti-Flt1, in vivo, was evaluated for inhibition of tumor growth and metastasis in VEGF-secreting cancer cell-implanted mice by s.c. injections of the peptide. RESULTS: Here, we report on a short peptide that binds to VEGFR1 and prevents binding of VEGF. A hexapeptide, anti-Flt1 (Gly-Asn-Gln-Trp Phe-Ile or GNQWFI), was identified from peptide libraries. The anti-Flt1 peptide shows specificity toward binding to VEGFR1 and it inhibits binding of VEGF, placental growth factor (PlGF), and VEGF/PlGF heterodimer to VEGFR1. This peptide does not inhibit the proliferation of endothelial cells induced by VEGF and VEGF/PlGF heterodimer but it effectively blocks VEGF-induced migration of endothelial cells and their capacity to form capillary-like structures on fibrin gel-based in vitro angiogenesis system. Furthermore, growth and metastasis of VEGF-secreting tumor cells were also significantly inhibited by s.c. injections of anti-Flt1 peptide in nude mice. Accordingly, VEGF-induced migration and capillary formation are mediated through VEGFR1, and these processes may play an important role in the growth and metastasis of VEGF-secreting tumors. CONCLUSIONS: We show that a peptide (anti-Flt1) specific for VEGFR1 inhibits growth and metastasis of tumor that secretes VEGF. The effects on endothelial cell functions, in vitro, indicate that the anticancer activity of anti-Flt1 peptide with reduced blood vessel density could also be due to the blocking of VEGFR1-mediated endothelial cell migration and tube formation. Although the effects of anti-Flt1 peptide still remain to be further characterized, the receptor 1-specific peptide antagonist, anti-Flt1, has potential as a therapeutic agent for various angiogenesis-related diseases, especially cancer. PMID- 15814645 TI - A phase I clinical, pharmacologic, and biologic study of thrombopoietin and granulocyte colony-stimulating factor in children receiving ifosfamide, carboplatin, and etoposide chemotherapy for recurrent or refractory solid tumors: a Children's Oncology Group experience. AB - PURPOSE: Ifosfamide, carboplatin, and etoposide (ICE) are associated with grade III/IV dose-limiting thrombocytopenia. The Children's Oncology Group conducted a phase I dose escalation, pharmacokinetic, and biological study of recombinant human thrombopoietin (rhTPO) after ICE in children with recurrent/refractory solid tumors (CCG-09717) to assess the toxicity and maximum tolerated dose of rhTPO administered at 1.2, 2.4, or 3.6 microg/kg per dose. EXPERIMENTAL DESIGN: Children received ifosfamide 1,800 mg/m2 on days 0 to 4, carboplatin 400 mg/m2 on days 0 to 1, and etoposide 100 mg/m2 on days 0 to 4. rhTPO was administered i.v. on days +4, +6, +8, +10, and +12 at 1.2, 2.4, or 3.6 microg/kg per dose. RESULTS: rhTPO was well tolerated and maximum tolerated dose was not reached. Median time to platelet recovery > or =100,000/microL of rhTPO at 1.2, 2.4, and 3.6 microg/kg/d was 24 days (22-24 d), 25 days (23-29 d), and 22 days (16-37 d), respectively. Patients required a median of 2 days of platelet transfusions (0-7 days). Mean (+/- SD) rhTPO maximum serum concentrations were 63.3 +/- 9.7 and 89.3 +/- 15.7 ng/mL and terminal half-lives were 47 +/- 13 and 64 +/- 42 hours after 2.4 and 3.6 microg/kg/d, respectively. There was a significant increase in colony-forming unit megakaryocyte upon WBC count recovery. CONCLUSIONS: rhTPO was well tolerated. Time to hematologic recovery and median number of platelet transfusions seem to be improved compared with historical controls receiving ICE + granulocyte colony-stimulating factor (CCG-0894). PMID- 15814647 TI - A small interfering RNA targeting vascular endothelial growth factor inhibits Ewing's sarcoma growth in a xenograft mouse model. AB - Angiogenesis plays an essential role in tumor growth and metastasis and is a promising therapeutic target for cancer. Vascular endothelial growth factor (VEGF) is a key regulator in vasculogenesis as well as in angiogenesis. TC71 human Ewing's sarcoma cells overexpress VEGF, with a shift in isoform production from membrane-bound VEGF189 to the more soluble VEGF165. Transfection of TC71 cells with a vector-based VEGF targeted small interfering RNA expression system (VEGFsi) inhibited VEGF165 expression by 80% and VEGF165 protein production by 98%, with no alteration in VEGF189 expression. Human microvascular endothelial cell proliferation and migration induced by conditioned medium from VEGFsi transfected TC71 cells was significantly less than that induced by conditioned medium from TC71 cells and control vector-transfected TC71 cells. Furthermore, after s.c. injection into athymic nu/nu mice, the tumor growth of VEGFsi expressing TC71 cells was significantly less than that of parental or control vector-transfected cells. Vessel density as assessed by CD31 immunohistochemical analysis and VEGF165 expression as assessed by Northern blotting were also decreased. Intratumor gene therapy with polyethylenimine/VEGFsi also resulted in tumor growth suppression. When inoculated into the tibias of nude mice, VEGFsi expressing TC71 cells induced osteolytic bone lesions that were less severe than those induced by control groups. These data suggest that targeting VEGF165 may provide a therapeutic option for Ewing's sarcoma. PMID- 15814648 TI - Caspase-dependent apoptosis induction by phenethyl isothiocyanate, a cruciferous vegetable-derived cancer chemopreventive agent, is mediated by Bak and Bax. AB - PURPOSE: The present study was undertaken to gain insights into the molecular mechanism of apoptosis induction by phenethyl isothiocyanate (PEITC) using prostate cancer cell lines derived from transgenic adenocarcinoma mouse prostate (TRAMP) mice (TRAMP-C1 and TRAMP-C2). EXPERIMENTAL DESIGN AND RESULTS: The viability of TRAMP-C1 and TRAMP-C2 cells was reduced significantly in the presence of PEITC in a concentration-dependent manner as determined by sulforhodamine B and trypan blue dye exclusion assays. Treatment of TRAMP-derived cells with PEITC revealed features characteristic of apoptosis induction, including appearance of subdiploid cells (determined by flow cytometry), cytoplasmic histone-associated DNA fragmentation (determined by an ELISA assay), and cleavage of caspase-3 (determined by immunoblotting). The PEITC-induced apoptosis in TRAMP-derived cells was associated with a marked increase in the level of proapoptotic protein Bak and/or a decrease in the levels of antiapoptotic protein Mcl-1 or Bcl-xL and disruption of mitochondrial membrane potential. The SV40 immortalized mouse embryonic fibroblasts derived from Bak and Bax double knockout mice were significantly more resistant to PEITC-induced DNA fragmentation compared with wild-type or Bak-/- mouse embryonic fibroblasts. The PEITC-induced apoptosis in both cell lines was significantly attenuated in the presence of caspase inhibitors zVAD-fmk, zLEHD-fmk, and zIETD-fmk. Oral administration of PEITC (9 or 12 micromol PEITC/d, Monday-Friday) significantly retarded growth of TRAMP-C1 xenografts in nude mice without causing weight loss or any other side effects. CONCLUSION: The results of the present study indicate that caspase-dependent apoptosis by PEITC is mediated by Bak and Bax proteins. PMID- 15814649 TI - Methionine aminopeptidase 2 inhibition is an effective treatment strategy for neuroblastoma in preclinical models. AB - Tumor vascularity is correlated with an aggressive disease phenotype in neuroblastoma, suggesting that angiogenesis inhibitors may be a useful addition to current therapeutic strategies. We previously showed that the antiangiogenic compound TNP-470, an irreversible methionine aminopeptidase 2 (MetAP2) inhibitor, suppressed local and disseminated human neuroblastoma growth rates in murine models but had significant associated toxicity at the effective dose. We have recently shown that a novel, reversible MetAP2 inhibitor, A-357300, significantly inhibits CHP-134-derived neuroblastoma s.c. xenograft growth rate with a treatment-to-control (T/C) ratio at day 24 of 0.19 (P < 0.001) without toxicity. We now show that the combination of A-357300 with cyclophosphamide at the maximal tolerated dose sustained tumor regression with a T/C at day 48 of 0.16 (P < 0.001) in the CHP-134 xenograft model. A-357300 also significantly inhibited establishment and growth rate of hematogenous metastatic deposits following tail vein inoculation of CHP-134 cells and increased overall survival (P = 0.021). Lastly, A-357300 caused regression of established tumors in a genetically engineered murine model with progression-free survival in five of eight mice (P < 0.0001). There was no evidence of toxicity. These data show that MetAP2 may be an important molecular target for high-risk human neuroblastomas. We speculate that the growth inhibition may be through both tumor cell intrinsic and extrinsic (antiangiogenic) mechanisms. The potential for a wide therapeutic index may allow for treatment strategies that integrate MetAP2 inhibition with conventional cytotoxic compounds. PMID- 15814650 TI - Functional expression of the angiotensin II type 1 receptor in human ovarian carcinoma cells and its blockade therapy resulting in suppression of tumor invasion, angiogenesis, and peritoneal dissemination. AB - PURPOSE: Angiotensin II is a bioactive peptide of the renin-angiotensin system, acting not only as a vasoconstrictor but also as a growth promoter via angiotensin II type 1 receptors (AT1R). The present study examined AT1R expression in human ovarian carcinoma and attempted to determine whether AT1R blocker could suppress the tumor progression. EXPERIMENTAL DESIGN: Expression of AT1R, vascular endothelial growth factor (VEGF), and CD34 was immunohistochemically analyzed in ovarian tumor tissues (n=99). Effects of AT1R blocker on invasive potential and VEGF secretion in ovarian cancer cells were examined in vitro. Effects of AT1R blocker in vivo were evaluated in a mouse model of peritoneal carcinomatosis. RESULTS: AT1R was expressed in 57 of 67 (85%) invasive ovarian adenocarcinomas and 12 of 18 (66%) borderline malignant tumors but in only 2 of 14 (14%) benign cystadenomas. In invasive carcinomas, VEGF expression intensity and intratumor microvessel density were significantly higher in cases that were strongly positive for AT1R (n = 37) compared with those in cases weakly positive (n = 20) or negative (n = 10) for AT1R. Angiotensin II significantly enhanced the invasive potential and VEGF secretion in AT1R-positive SKOV-3 ovarian cancer cells, both of which were completely inhibited by the AT1R blocker candesartan. Administration of candesartan into SKOV-3-transplanted athymic mice resulted in the reduction of peritoneal dissemination, decreased ascitic VEGF concentration, and suppression of tumor angiogenesis. CONCLUSIONS: AT1R is functionally expressed in ovarian carcinoma and involved in tumor progression and angiogenesis. AT1R blockade therapy may become a novel and promising strategy for ovarian cancer treatment. PMID- 15814651 TI - Preclinical evaluation of the pharmacodynamic properties of 2,5-diaziridinyl-3 hydroxymethyl-6-methyl-1,4-benzoquinone. AB - PURPOSE: The purpose of our study was to investigate the cellular accumulation, DNA cross-linking ability, and cellular toxicity of RH1 (2,5-diaziridinyl-3 [hydroxymethyl[-6-methyl-1,4-benzoquinone), a novel DNA alkylating agent currently in clinical trials. In addition, the in vivo efficacy of RH1 formulated in different vehicles was also compared. EXPERIMENTAL DESIGN: RH1 is activated by the two-electron reducing enzyme NQO1 [NADPH:quinone oxidoreductase] forming a potent cytotoxic agent that cross-links DNA. We have used whole blood, cell lines, and primary explanted tumor cultures to measure both the cellular accumulation, DNA cross-linking, and cytotoxicity of RH1. Furthermore, the pharmacokinetic and pharmacodynamic characteristics of RH1 formulated in different vehicles were measured in vivo using the validated comet-X assay in mice bearing human tumor xenografts. RESULTS: Accumulation of RH1 was shown to be both time and concentration dependent, reaching a maximum after 2 hours and correlated well with DNA cross-linking measurements. DNA cross-linking in vitro could be detected at low (1-10 nmol/L) concentrations after as little as 2 hours exposure. In primary tumor cultures, RH1 induces much higher levels of DNA cross links at lower doses than either mitomycin C or cisplatin. In vivo efficacy testing using polyvinyl pyrrolidone, saline, or cyclodextrin as vehicles showed DNA cross-links readily detectable in all tissues examined and was enhanced when given in cyclodextrin compared with polyvinyl pyrrolidone or saline. CONCLUSIONS: RH1 represents a potent bioreductive anticancer drug, which may prove effective in the treatment of cancers, particularly those that overexpress NQO1. DNA cross linking can be reliably measured in tissue using the validated comet-X assay. PMID- 15814652 TI - Disruption of fibroblast growth factor signal pathway inhibits the growth of synovial sarcomas: potential application of signal inhibitors to molecular target therapy. AB - PURPOSE: Synovial sarcoma is a soft tissue sarcoma, the growth regulatory mechanisms of which are unknown. We investigated the involvement of fibroblast growth factor (FGF) signals in synovial sarcoma and evaluated the therapeutic effect of inhibiting the FGF signal. EXPERIMENTAL DESIGN: The expression of 22 FGF and 4 FGF receptor (FGFR) genes in 18 primary tumors and five cell lines of synovial sarcoma were analyzed by reverse transcription-PCR. Effects of recombinant FGF2, FGF8, and FGF18 for the activation of mitogen-activated protein kinase (MAPK) and the growth of synovial sarcoma cell lines were analyzed. Growth inhibitory effects of FGFR inhibitors on synovial sarcoma cell lines were investigated in vitro and in vivo. RESULTS: Synovial sarcoma cell lines expressed multiple FGF genes especially those expressed in neural tissues, among which FGF8 showed growth stimulatory effects in all synovial sarcoma cell lines. FGF signals in synovial sarcoma induced the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and p38MAPK but not c-Jun NH2-terminal kinase. Disruption of the FGF signaling pathway in synovial sarcoma by specific inhibitors of FGFR caused cell cycle arrest leading to significant growth inhibition both in vitro and in vivo. Growth inhibition by the FGFR inhibitor was associated with a down regulation of phosphorylated ERK1/2 but not p38MAPK, and an ERK kinase inhibitor also showed growth inhibitory effects for synovial sarcoma, indicating that the growth stimulatory effect of FGF was transmitted through the ERK1/2. CONCLUSIONS: FGF signals have an important role in the growth of synovial sarcoma, and inhibitory molecules will be of potential use for molecular target therapy in synovial sarcoma. PMID- 15814653 TI - Effect of YM529 on a model of mandibular invasion by oral squamous cell carcinoma in mice. AB - PURPOSE: This study examined the mechanisms of osteoclast-mediated bone invasion in a model of oral squamous cell carcinoma (OSCC). C3H/HeN mice were inoculated with SCC VII cells into the masseter region to establish an animal model of mandibular invasion by OSCC. EXPERIMENTAL DESIGN: The mice were divided into three groups: a control group, given daily s.c. injections of saline; group 1, given 2 microg per mouse per day of the bisphosphonate YM529; and group 2, given 10 microg per mouse per day of YM529. After 3 weeks of treatment, the lesions were studied by micro-computed tomography. After tartrate-resistant acid phosphatase (TRAP) staining, the osteoclasts were easily identified, and the percentages of the area occupied by osteoclasts were calculated by computer for each sample. The tumors were analyzed by RT-PCR to determine the mRNA expression of interleukin-6 (IL-6), parathyroid hormone-related protein (PTHrP), tumor necrosis factor-alpha (TNF-alpha), receptor activator of nuclear factor-kappaB (RANK), RANK ligand (RANKL), and osteoprotegerin. RESULTS: SCC VII cells rapidly multiplied in the masseter muscle of the mice. Bone invasion was evident only in the control group on micro-computed tomography. On TRAP-stained slices, the percentages of osteoclasts in groups 1 and 2 were significantly lower than that in the control group. The mRNA expressions of IL-6, PTHrP, THF-alpha, and RANK decreased as the concentration of YM529 increased. CONCLUSIONS: We conclude that various cancer-derived cytokines play important roles in the invasion of bone by OSCC. YM529, a third-generation bisphosphonate, can suppress osteoclast-mediated bone invasion by OSCC. The mechanism of this effect might involve inhibition of cytokines such as IL-6, PTHrP, TNF-alpha, and RANK by YM529. PMID- 15814654 TI - Potential antagonism of tubulin-binding anticancer agents in combination therapies. AB - ZD6126 is a vascular targeting agent, developed for the treatment of solid tumors. In vivo, ZD6126 is rapidly converted into the tubulin-binding agent N acetylcolchinol. We have previously reported that in vitro N-acetylcolchinol disrupts microtubules and induces rapid changes in endothelial cell morphology, which in a tumor would lead to a rapid loss of tumor vessel integrity and subsequent extensive tumor necrosis. The aim of this study was to investigate the effect of cytotoxic antineoplastic drugs-cisplatin, doxorubicin, vincristine, paclitaxel, and docetaxel-on endothelial cell response to N-acetylcolchinol. We found that cisplatin and doxorubicin did not interfere with the ability of N acetylcolchinol to cause morphologic changes in human umbilical vein endothelial cells, whereas vincristine showed additive effects. In contrast, the microtubule stabilizing agents paclitaxel (1-10 micromol/L) and docetaxel (0.1-1 micromol/L) prevented the morphologic changes induced by N-acetylcolchinol in human umbilical vein endothelial cells. The effect was observed when cells were exposed to paclitaxel and N-acetylcolchinol together or when paclitaxel was given shortly before N-acetylcolchinol. Paclitaxel and N-acetylcolchinol interacted at the level of microtubule organization, as shown in immunofluorescence analysis of the cytoskeleton. The protective effect was reversible because 4 hours after paclitaxel wash out, cells recovered the sensitivity to N-acetylcolchinol. In vivo, pretreatment of mice with paclitaxel inhibited the vascular targeting activity of ZD6126 on newly formed vessels in the Matrigel plug assay and ZD6126 induced necrosis in tumors. These findings indicate that paclitaxel, depending on the timing and schedule of administration, can affect the vascular targeting activity of ZD6126, which may have an effect on the optimal scheduling of therapies based on the combined use of microtubule-stabilizing and microtubule destabilizing agents. PMID- 15814655 TI - Advantage of a residualizing iodine radiolabel in the therapy of a colon cancer xenograft targeted with an anticarcinoembryonic antigen monoclonal antibody. AB - PURPOSE: A disadvantage of conventionally radioiodinated monoclonal antibodies (mAb) for cancer therapy is the short retention time of the radionuclide within target cells. To address this issue, we recently developed a method in which radioiodine is introduced onto antibodies using an adduct consisting of a nonmetabolizable peptide attached to the aminopolycarboxylate diethylenetriaminepentaacetic acid, designated IMP-R4. This adduct causes the radioiodine to become trapped in lysosomes following antibody catabolism. Clinical-scale production of 131I-IMP-R4-labeled antibodies is possible using a recently developed facile method. EXPERIMENTAL DESIGN: The properties of 131I-IMP R4-labeled anticarcinoembryonic antigen (CEA) humanized mAb hMN-14 were compared with the directly radioiodinated hMN-14 (131I-hMN-14) in CEA-expressing human colon cancer cell lines, LoVo and LS174T, and in nude mice bearing established LoVo tumor xenografts. RESULTS: 125I-IMP-R4-hMN-14 retention in the cell lines was significantly increased (61.5% after 3 days) compared with 125I-hMN-14. In vivo, a significant improvement in tumor accretion of radiolabel was obtained using 131I-IMP-R4-hMN-14, which led to a marked improvement in therapeutic efficacy. Eight weeks post-treatment, mean tumor volumes were 0.16 +/- 0.19 and 1.99 +/- 1.35 cm3 in mice treated with 131I-IMP-R4-hMN-14 and 131I-hMN-14, respectively, with complete remissions observed in 27% of mice treated with 131I IMP-R4-hMN-14 and none using 131I-hMN-14. CONCLUSION: 131I-IMP-R4-hMN-14 provides a significant therapeutic advantage in comparison to the conventionally 131I labeled antibody. The ability of this labeling method to lend itself to clinical scale labeling, the broad applicability of a humanized anti-CEA mAb for CEA expressing cancers, and the clinical benefits of radioimmunotherapy with anti-CEA mAb shown recently for small-volume and minimal residual disease combine to make 131I-IMP-R4-hMN-14 a promising new agent for radioimmunotherapy. PMID- 15814656 TI - (-)-Epigallocatechin gallate and polyphenon E inhibit growth and activation of the epidermal growth factor receptor and human epidermal growth factor receptor-2 signaling pathways in human colon cancer cells. AB - PURPOSE: (-)-Epigallocatechin gallate (EGCG) inhibits activation of the epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor-2 (HER2) and multiple downstream signaling pathways in cancer cell lines. In this study we compared the cellular and molecular effects of EGCG with a well-standardized decaffeinated green tea catechin mixture Polyphenon E (Poly E) on human colon cancer cell lines. EXPERIMENTAL DESIGN AND RESULTS: Both EGCG and Poly E preferentially inhibited growth of the Caco2, HCT116, HT29, SW480, and SW837 colon cancer cells when compared with the FHC normal human fetal colon cell line. The EGFR and HER2 proteins were overexpressed and constitutively activated in all of the colon cancer cell lines when compared with the FHC cell line. Treatment of HT29 cells with EGCG or Poly E caused an increase of cells in G1 and induced apoptosis. Both EGCG and Poly E caused a decrease in the phosphorylated forms of EGFR and HER2 proteins, and subsequently caused a decrease in the phosphorylated forms of the extracellular signal-regulated kinase and Akt proteins. Similar effects of these compounds were seen when the cells were stimulated with transforming growth factor alpha. Reporter assays indicated that both EGCG and Poly E inhibited the transcriptional activity of the activator protein 1 (AP-1), c-fos, nuclear factor kappaB, and cyclin D1 promoters. The combination of only 1 microg/mL of epicatechin plus 10 microg/mL of EGCG displayed synergistic effects on growth inhibition and induction of apoptosis. Furthermore, when treatment was prolonged for 96 hours, 1 microg/mL of EGCG or Poly E was sufficient to inhibit growth, reduce activation of EGFR and HER2, and induce apoptosis. CONCLUSION: Our findings suggest that EGCG or Poly E may be useful in the chemoprevention and/or treatment of colon cancer. Poly E contains about 60% EGCG, yet pure EGCG and Poly E had similar potencies (expressed as microg/ml). Poly E may be preferable because it is easier to prepare and this mixture of catechins may exert synergistic effects. PMID- 15814657 TI - O6-methylguanine-DNA methyltransferase, O6-benzylguanine, and resistance to clinical alkylators in pediatric primary brain tumor cell lines. AB - PURPOSE: Primary brain tumors are the leading cause of cancer death in children. Our purpose is (a) to assess the contribution of the DNA repair protein O6 methylguanine-DNA methyltransferase (MGMT) to the resistance of pediatric brain tumor cell lines to clinical alkylating agents and (b) to evaluate variables for maximal potentiation of cell killing by the MGMT inhibitor O6-benzylguanine, currently in clinical trials. Few such data for pediatric glioma lines, particularly those from low-grade tumors, are currently available. EXPERIMENTAL DESIGN: We used clonogenic assays of proliferative survival to quantitate cytoxicity of the chloroethylating agent 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and the methylating agent temozolomide in 11 glioma and five medulloblastoma lines. Twelve lines are newly established and characterized here, nine of them from low-grade gliomas including pilocytic astrocytomas. RESULTS: (a) MGMT is a major determinant of BCNU resistance and the predominant determinant of temozolomide resistance in both our glioma and medulloblastoma lines. On average, O(6)-benzylguanine reduced LD10 for BCNU and temozolomide, 2.6 and 26-fold, respectively, in 15 MGMT-expressing lines. (b) O6-Benzylguanine reduced DT (the threshold dose for killing) for BCNU and temozolomide, 3.3- and 138-fold, respectively. DT was decreased from levels higher than, to levels below, clinically achievable plasma doses for both alkylators. (c) Maximal potentiation by O6-benzylguanine required complete and prolonged suppression of MGMT. CONCLUSIONS: Our results support the use of O6-benzylguanine to achieve full benefit of alkylating agents, particularly temozolomide, in the chemotherapy of pediatric brain tumors. PMID- 15814659 TI - Microregional expression of glucose transporter-1 and oxygenation status: lack of correlation in locally advanced cervical cancers. AB - PURPOSE: Glucose transporter-1 (GLUT-1), a target gene of hypoxia-inducible factor-1, has been considered a candidate endogenous marker of tumor hypoxia. Expression of GLUT-1 may also serve as an indicator for the induction of the transcriptional response to hypoxia, which has been linked to enhanced proliferation, resistance to therapy, and metastatic propagation of cancer cells. Overexpression of GLUT-1 has been shown to correlate with poor prognosis in several tumor entities, among them cancers of the uterine cervix. The validity of these hypotheses is investigated. EXPERIMENTAL DESIGN: The expression of GLUT-1 was assessed in 80 biopsies of Eppendorf oxygenation measurement tracks from locally advanced cervical cancers in 47 patients using immunohistochemistry. RESULTS: No correlation was found between the expression of GLUT-1 and oxygenation variables (median pO2, HF 2.5 and HF 5). Expression of GLUT-1 was found greater in larger tumors (P = 0.0001) and to exhibit a linear increase with Federation Internationale de Gynecologie et d' Obstetrique stage (P = 0.002). Overall survival (P = 0.004) and recurrence-free survival (P = 0.007) were significantly shorter for patients with expression of GLUT-1. In the subgroup of patients treated with surgery, this effect on prognosis was not independent when pT stage or pN stage were included in a multivariate Cox proportional hazards model. CONCLUSIONS: The suitability of GLUT-1 as an endogenous marker of tumor hypoxia seems questionable. The association with prognosis may partially depend on confounding factors. PMID- 15814658 TI - Myc down-regulation sensitizes melanoma cells to radiotherapy by inhibiting MLH1 and MSH2 mismatch repair proteins. AB - PURPOSE: Melanoma patients have a very poor prognosis with a response rate of <1% due to advanced diagnosis. This type of tumor is particularly resistant to conventional chemotherapy and radiotherapy, and the surgery remains the principal treatment for patients with localized melanoma. For this reason, there is particular interest in the melanoma biological therapy. EXPERIMENTAL DESIGN: Using two p53 mutant melanoma models stably expressing an inducible c-myc antisense RNA, we have investigated whether Myc protein down-regulation could render melanoma cells more susceptible to radiotherapy, reestablishing apoptotic p53-independent pathway. In addition to address the role of p53 in the activation of apoptosis, we studied the effect of Myc down-regulation on radiotherapy sensitivity also in a p53 wild-type melanoma cell line. RESULTS: Myc down regulation is able per se to induce apoptosis in a fraction of the cell population (approximately 40% at 72 hours) and in combination with gamma radiation efficiently enhances the death process. In fact, approximately 80% of apoptotic cells are evident in Myc down-regulated cells exposed to gamma radiation for 72 hours compared with approximately 13% observed after only gamma radiation treatment. Consistent with the enhanced apoptosis is the inhibition of the MLH1 and MSH2 mismatch repair proteins, which, preventing the correction of ionizing radiation mismatches occurring during DNA replication, renders the cells more prone to radiation-induced apoptosis. CONCLUSIONS: Data herein reported show that Myc down-regulation lowers the apoptotic threshold in melanoma cells by inhibiting MLH1 and MSH2 proteins, thus increasing cell sensitivity to gamma radiation in a p53-independent fashion. Our results indicate the basis for developing new antitumoral therapeutic strategy, improving the management of melanoma patients. PMID- 15814660 TI - Tirapazamine cytotoxicity for neuroblastoma is p53 dependent. AB - Relapse of neuroblastoma commonly occurs in hypoxic tissues, and is associated with an acquired and sustained high-level drug resistance, often due to p53 loss of function. Abrogating p53 function with HPV 16 E6 transduction in drug sensitive neuroblastoma cell lines caused high-level drug resistance. Tirapazamine (TPZ) is a bioreductive agent that forms a toxic free radical in hypoxia. We determined in six neuroblastoma cell lines the cytotoxicity of TPZ using DIMSCAN, a digital imaging fluorescence assay, apoptosis and mitochondrial membrane potential (DeltaPsim) by flow cytometry, and protein expression by immunoblotting. TPZ exhibited high cytotoxicity, especially in hypoxia (2% O2), for all four p53-functional neuroblastoma cell lines, achieving >3 logs of cell kill (LC99 < or = 0.7 microg/mL). In p53-nonfunctional neuroblastoma cell lines, all TPZ LC99 values were >3.0 microg/mL (average clinically achievable level). TPZ (24 hours) induced apoptosis in >46% of cells in p53-functional cell lines but failed to cause apoptosis in p53 nonfunctional cell lines. Induction of p53 and p21 expression by TPZ was observed in a p53-functional cell line (SMS-SAN) but not in a p53-nonfunctional cell line (CHLA-90). Significant DeltaPsim loss and glutathione (GSH) depletion in response to TPZ was observed in p53-functional cell lines (SMS-SAN, SMS-SAN EV, and CHLA-15) but not in p53-nonfunctional cell lines (SMS-SAN E6 and CHLA-90). N-Acetylcysteine inhibited TPZ-mediated DeltaPsim loss and GSH depletion, but neither N-acetylcysteine nor Boc-d-fmk inhibited apoptosis caused by TPZ. In response to TPZ, DeltaPsim loss preceded apoptosis. Thus, TPZ cytotoxicity for neuroblastoma cell lines in hypoxia occurred via a p53 dependent mitochondrial pathway that caused induction of p53 and p21, DeltaPsim decrease, GSH depletion, and apoptosis. These data further define the mechanism of action of TPZ and suggest that as a single agent, TPZ would only have clinical activity against p53-functional neuroblastomas. PMID- 15814661 TI - The dl1520 virus is found preferentially in tumor tissue after direct intratumoral injection in oral carcinoma. PMID- 15814662 TI - Increased IgE but reduced Th2-type inflammation in vitamin D receptor-deficient mice. PMID- 15814663 TI - TLR signaling in the gut in health and disease. AB - The human intestine has evolved in the presence of diverse enteric microflora. TLRs convert the recognition of pathogen-associated molecules in the gut into signals for anti-microbial peptide expression, barrier fortification, and proliferation of epithelial cells. Healing of injured intestinal epithelium and clearance of intramucosal bacteria require the presence of intact TLR signaling. Nucleotide oligomerization domain (Nod)1 and Nod2 are additional pattern recognition receptors that are required for defense against invasive enteric pathogens. Through spatial and functional localization of TLR and Nod molecules, the normal gut maintains a state of controlled inflammation. By contrast, patients with inflammatory bowel disease demonstrate inflammation in response to the normal flora. A subset of these patients carry polymorphisms in TLR and CARD15/NOD2 genes. A better understanding of the delicate regulation of TLR and Nod molecules in the gut may lead to improved treatment for enteric infections and idiopathic inflammatory bowel diseases. PMID- 15814664 TI - Death of a dogma or enforcing the artificial: monomeric IgE binding may initiate mast cell response by inducing its receptor aggregation. AB - Several recent reports have suggested that binding monomeric IgE (mIgE) to its type 1 receptor, Fc epsilon RI, on mast cells induces important responses. These observations contradict the notion that it is the aggregation of this receptor that is essential for initiating mast cell response. In the present study, we suggest that the most probable causes for the reported observations are the experimental protocol used combined with the high expression levels of the Fc epsilon RI by mast cells. Specifically, we suggest using the published data and physicochemical calculations that the exceptionally high number of cell surface Fc epsilon RI-bound monoclonal IgE yields, in the two-dimensions of the cells' membranes, a situation where even a low affinity of these mIgE for epitopes on their own structure or on another cell surface component may lead to their aggregation. Hence, we hypothesize that the reported response to mIgE binding is a result of such an Fc epsilon RI-IgE induced aggregation. PMID- 15814665 TI - Type I IFNs provide a third signal to CD8 T cells to stimulate clonal expansion and differentiation. AB - In this study, we show that IFN-alpha beta can have a direct role in linking innate and adaptive responses by providing the "third signal" needed by naive CD8 T cells responding to Ag and costimulatory ligands. Stimulation of CD8 T cells in the absence of a third signal leads to proliferation, but clonal expansion is limited by poor survival and effector functions do not develop. We show that IFN alpha beta can provide the third signal directly to CD8 T cells via a STAT4 dependent pathway to stimulate survival, development of cytolytic function, and production of IFN-gamma. Provision of the third signal by either IFN-alpha beta or IL-12 results in regulation of the expression of a number of genes, including several that encode proteins critical for effector function. PMID- 15814666 TI - Fas ligand (CD178) cytoplasmic tail is a positive regulator of Fas ligand mediated cytotoxicity. AB - The cytotoxic function of CD178 (Fas ligand (FasL)) is critical to the maintenance of peripheral tolerance and immune-mediated tissue pathology. The active site of FasL resides at the FasL extracellular region (FasL(Ext)) and it functions through binding/cross-linking Fas receptor on target cells. In this study, we report that FasL(Ext)-mediated cytotoxicity is regulated by the FasL cytoplasmic tail (FasL(Cyt)). Deleting the N-terminal 2-70 aa (delta70) or N terminal 2-33 aa (delta33) reduced the cytotoxic strength as much as 30- to 100 fold. By contrast, change in the cytotoxic strength was not observed with FasL deleted of the proline-rich domains (45-74 aa, delta PRD) in the FasL(Cyt). Our study identifies a novel function of FasL(Cyt) and demonstrates that FasL(2-33), a sequence unique to FasL, is critically required for the optimal expression of FasL(Ext)-mediated cytotoxicity. PMID- 15814667 TI - Itk is not essential for CD28 signaling in naive T cells. AB - Itk, a member of the Tec family of tyrosine kinases, is critical for TCR signaling, leading to the activation of phospholipase C gamma1. Early biochemical studies performed in tumor cell lines also implicated Itk in CD28 signaling. These data were complemented by functional studies on primary Itk-/- T cells that suggested a negative role for Itk in CD28 signaling. In this report, we describe a thorough analysis of CD28-mediated responses in T cells lacking Itk. Using purified naive CD4+ T cells from Itk-/- mice, we examine a range of responses dependent on CD28 costimulation. We also analyze Akt and glycogen synthase kinase 3beta phosphorylation in response to stimulation of CD28 alone. Overall, these experiments demonstrate that CD28 signaling, as well as CD28-mediated costimulation of TCR signaling, function efficiently in the absence of Itk. These findings indicate that Itk is not essential for CD28 signaling in primary naive CD4+ T cells. PMID- 15814668 TI - NKG2D is a costimulatory receptor for human naive CD8+ T cells. AB - In humans, all alpha beta CD8+ T cells express NKG2D, but in mouse, it is only expressed by activated and memory CD8+ T cells. We purified human naive CD8+ T cells to show that NKG2D serves as a costimulatory receptor for TCR induced Ca2+ mobilization and proliferation. The resulting effector cells are skewed toward a type 1 phenotype and produce high levels of IFN-gamma and TNF-alpha. NKG2D ligands, MHC class I chain-related (MIC)A, MICB, and UL16-binding proteins are expressed on the proliferating cells and NKG2D is down-regulated. The addition of the homeostatic cytokines IL-7 and IL-15 to the culture medium not only enhances proliferation but also counteracts the down-regulation of NKG2D, more so than the addition of IL-2. These results indicate that NKG2D can regulate the priming of human naive CD8+ T cells, which may provide an alternative mechanism for potentiating and channeling the immune response. PMID- 15814669 TI - Generation of high-affinity antibody against T cell-dependent antigen in the Ganp gene-transgenic mouse. AB - Generation of high-affinity Ab is impaired in mice lacking germinal center associated DNA primase (GANP) in B cells. In this study, we examined the effect of its overexpression in ganp transgenic C57BL/6 mice (Ganp(Tg)). Ganp(Tg) displayed normal phenotype in B cell development, serum Ig levels, and responses against T cell-independent Ag; however, it generated the Ab with much higher affinity against nitrophenyl-chicken gammaglobulin in comparison with C57BL/6. To further examine the affinity increase, we established hybridomas producing high affinity mAbs and compared their affinities using BIAcore. C57BL/6 generated high affinity anti-nitrophenyl mAbs (K(D) approximately 2.50 x 10(-7) M) of IgG1/lambda1 and contained the V(H)186.2 region with W33L mutation. Ganp(Tg) generated much higher affinity (K(D) > 1.57 x 10(-9) M) by usage of V(H)186.2 as well as noncanonical V(H)7183 regions. Ganp(Tg) also generated exceptionally high affinity anti-HIV-1 (V3 peptide) mAbs (K(D) > 9.90 x 10(-11) M) with neutralizing activity. These results demonstrated that GANP is involved in V region alteration generating high-affinity Ab. PMID- 15814670 TI - Regulation of highly cytokinergic IgE-induced mast cell adhesion by Src, Syk, Tec, and protein kinase C family kinases. AB - Mast cells play a critical role in IgE-dependent immediate hypersensitivity. Recent studies have shown that, contrary to the traditional view, binding of monomeric IgE to Fc epsilon RI results in a number of biological outcomes in mast cells, including survival. However, IgE molecules display heterogeneity in inducing cytokine production; highly cytokinergic (HC) IgEs cause extensive Fc epsilon RI aggregation, which leads to potent enhancement of survival and other activation events, whereas poorly cytokinergic (PC) IgEs can do so inefficiently. The present study demonstrates that HC, but not PC, IgEs can efficiently induce adhesion and spreading of mouse mast cells on fibronectin-coated plates in slow and sustained kinetics. HC IgE-induced adhesion through beta1 and beta7 integrins promotes survival, IL-6 production, and DNA synthesis. Importantly, we have identified Lyn and Syk as requisite tyrosine kinases and Hck, Btk, and protein kinase C theta as contributory kinases in HC IgE-induced adhesion and spreading, whereas protein kinase C epsilon plays a negative role. Consistent with these results, Lyn, Syk, and Btk are activated in HC IgE-stimulated cells in a slower but more sustained manner, compared with cells stimulated with IgE and Ag. Thus, binding of HC IgEs to Fc epsilon RI induces adhesion of mast cells to fibronectin by modulating cellular activation signals in a unique fashion. PMID- 15814671 TI - Antinuclear antigen B cells that down-regulate surface B cell receptor during development to mature, follicular phenotype do not display features of anergy in vitro. AB - We previously demonstrated that B cells expressing a transgenic BCR with "dual reactivity" for the hapten arsonate and nuclear autoantigens efficiently complete development to follicular phenotype and stably reside in follicles in vivo. These B cells express very low levels of surface IgM and IgD, suggesting that they avoid central deletion and peripheral anergy by reducing their avidity for autoantigen via surface BCR (sBCR) down-regulation. Since a variety of states of B cell anergy have been previously described, a thorough examination of the functional capabilities of these B cells was required to test this hypothesis. In this study, we show that surface Ig cross-linking induces amounts of proximal BCR signaling in these B cells commensurate with their reduced sBCR levels. Functionally, however, they are comparable to nonautoreactive B cells in cell cycle progression, up-regulation of activation and costimulatory molecules, and Ab-forming cell differentiation when treated with a variety of stimuli in vitro. In addition, these B cells can efficiently process and present Ag and are capable of undergoing cognate interaction with naive TCR-transgenic T cells, resulting in robust IL-2 production. Together, these data reveal a lack of intrinsic anergy involving any known mechanism, supporting the idea that this type of antinuclear Ag B cell becomes indifferent to cognate autoantigen by down-regulating sBCR. PMID- 15814672 TI - Active tolerance induction and prevention of autoimmune diabetes by immunogene therapy using recombinant adenoassociated virus expressing glutamic acid decarboxylase 65 peptide GAD(500-585). AB - Tolerance induction of autoreactive T cells against pancreatic beta cell-specific autoantigens such as glutamic acid decarboxylase 65 (GAD65) and insulin has been attempted as a method to prevent autoimmune diabetes. In this study, we investigate whether adenoassociated virus (AAV) gene delivery of multiple immunodominant epitopes expressing GAD(500-585) could induce potent immune tolerance and persistently suppress autoimmune diabetes in NOD mice. A single muscle injection of 7-wk-old female NOD mice with rAAV/GAD(500-585) (3 x 10(11) IU/mouse) quantitatively reduced pancreatic insulitis and efficiently prevented the development of overt type I diabetes. This prevention was marked by the inactivation of GAD(500-585)-responsive T lymphocytes, the enhanced GAD(500-585) specific Th2 response (characterized by increased IL-4, IL-10 production, and decreased IFN-gamma production; especially elevated anti-GAD(500-585) IgG1 titer; and relatively unchanged anti-GAD(500-585) IgG2b titer), the increased secretion of TGF-beta, and the production of protective regulatory cells. Our studies also revealed that peptides 509-528, 570-585, and 554-546 in the region of GAD(500 585) played important roles in rAAV/GAD(500-585) immunization-induced immune tolerance. These data indicate that using AAV, a vector with advantage for therapeutic gene delivery, to transfer autoantigen peptide GAD(500-585), can induce immunological tolerance through active suppression of effector T cells and prevent type I diabetes in NOD mice. PMID- 15814673 TI - Treatment with nonmitogenic anti-CD3 monoclonal antibody induces CD4+ T cell unresponsiveness and functional reversal of established experimental autoimmune encephalomyelitis. AB - In vivo administration of anti-CD3 Ab induces both immune tolerance and undesirable side-effects resulting from nonspecific proinflammatory cytokine production. In the current study, we investigated the therapeutic potential of two structurally altered forms of the anti-CD3 Ab in ameliorating established experimental autoimmune encephalomyelitis. Administration of either a chimeric (NM-IgG3) or digestion product (NM-F(ab')2) form of the anti-CD3 Ab during established experimental autoimmune encephalomyelitis conferred significant protection from clinical disease progression and was associated with decreased Ag specific T cell proliferation, cytokine production, and CNS inflammation. Interestingly, while this protection correlated with an increase in the frequency of CD4(+)CD25(+) regulatory T cells, neither prior depletion of regulatory T cells nor anti-TGF-beta treatment abrogated the treatment's efficacy. Importantly, both treatments induced normal levels of intracellular Ca(2+)-flux, but significantly diminished levels of TCR signaling. Consequent to this decreased level of TCR-mediated signaling were alterations in the level of apoptosis and CD4+ T cell trafficking resulting in a profound lymphopenia. Collectively, these results indicate that nonmitogenic anti-CD3 directly induces a state of immune unresponsiveness in primed pathogenic autoreactive effector cells via mechanisms that may involve the induction of T cell tolerance, apoptosis, and/or alterations in cell trafficking. PMID- 15814674 TI - Expression profiling of murine double-negative regulatory T cells suggest mechanisms for prolonged cardiac allograft survival. AB - Recent studies have demonstrated that both mouse and human alpha beta TCR(+)CD3(+)NK1.1(-)CD4(-)CD8- double-negative regulatory T (DN Treg) cells can suppress Ag-specific immune responses mediated by CD8+ and CD4+ T cells. To identify molecules involved in DN Treg cell function, we generated a panel of murine DN Treg clones, which specifically kill activated syngeneic CD8+ T cells. Through serial cultivation of DN Treg clones, mutant clones arose that lost regulatory capacity in vitro and in vivo. Although all allogeneic cardiac grafts in animals preinfused with tolerant CD4/CD8 negative 12 DN Treg clones survived over 100 days, allograft survival is unchanged following infusion of mutant clones (19.5 +/- 11.1 days) compared with untreated controls (22.8 +/- 10.5 days; p < 0.001). Global gene expression differences between functional DN Treg cells and nonfunctional mutants were compared. We found 1099 differentially expressed genes (q < 0.025%), suggesting increased cell proliferation and survival, immune regulation, and chemotaxis, together with decreased expression of genes for Ag presentation, apoptosis, and protein phosphatases involved in signal transduction. Expression of 33 overexpressed and 24 underexpressed genes were confirmed using quantitative real-time PCR. Protein expression of several genes, including Fc epsilon RI gamma subunit and CXCR5, which are >50-fold higher, was also confirmed using FACS. These findings shed light on the mechanisms by which DN Treg cells down-regulate immune responses and prolong cardiac allograft survival. PMID- 15814675 TI - A sexual dimorphism in intrathymic sialylation survey is revealed by the trans sialidase from Trypanosoma cruzi. AB - Sialylation is emerging as an important issue in developing thymocytes and is considered among the most significant cell surface modifications, although its physiologic relevance is far from being completely understood. It is regulated by the concerted expression of sialyl transferases along thymocyte development. After in vivo administration of trans-sialidase, a virulence factor from the American trypanosomatid Trypanosoma cruzi that directly transfers the sialyl residue among macromolecules, we found that the alteration of the sialylation pattern induces thymocyte apoptosis inside the "nurse cell complex." This suggests a glycosylation survey in the development of the T cell compartment. In this study, we report that this thymocyte apoptosis mechanism requires the presence of androgens. No increment in apoptosis was recorded after trans sialidase administration in females or in antiandrogen-treated, gonadectomized, or androgen receptor mutant male mice. The androgen receptor presence was required only in the thymic epithelial cells as determined by bone marrow chimeric mouse approaches. The presence of the CD43 surface mucin, a molecule with a still undefined function in thymocytes, was another absolute requirement. The trans-sialidase-induced apoptosis proceeds through the TNF-alpha receptor 1 deathly signaling leading to the activation of the caspase 3. Accordingly, the production of the cytokine was increased in thymocytes. The ability of males to delete thymocytes altered in their sialylation pattern reveals a sexual dimorphism in the glycosylation survey during the development of the T cell compartment that might be related to the known differences in the immune response among sexes. PMID- 15814676 TI - Role for glycogen synthase kinase-3 in NK cell cytotoxicity and X-linked lymphoproliferative disease. AB - NK cells from individuals with X-linked lymphoproliferative (XLP) disease exhibit functional defects when stimulated through the NK receptor, 2B4 (CD244). These defects are likely a consequence of aberrant intracellular signaling initiated by mutations of the adaptor molecule SLAM-associated protein. In this report, we show that NK cells from individuals with XLP but not healthy individuals fail to phosphorylate and thereby inactivate glycogen synthase kinase-3 (GSK-3) following 2B4 stimulation. Lack of GSK-3 phosphorylation prevented the accumulation of the transcriptional coactivator beta-catenin in the cytoplasm and its subsequent translocation to the nucleus. Potential signaling pathways leading from 2B4 stimulation to GSK-3 phosphorylation were also investigated. Ligation of 2B4 resulted in the phosphorylation of the guanine nucleotide exchange factor, Vav-1, and subsequent activation of the GTP-binding protein Rac-1 (but not Ras) and the serine-threonine kinase Raf-1 in healthy but not XLP-derived NK cells. In addition, the activity of MEK-2 (but not MEK-1) was up-regulated, and Erk1/2 was phosphorylated in normal NK cells but not those from an individual with XLP suggesting that these proteins relay SLAM-associated protein-dependent signals from 2B4. Finally, inactivation of GSK-3 using a specific inhibitor of GSK-3beta increased the cytotoxicity and cytokine secretion of both healthy and XLP NK cells. These data indicate that the signaling of 2B4 in NK cells is mediated by GSK-3 and beta-catenin, possibly through a signal transduction pathway that involves Vav-1, Rac-1, Raf-1, MEK-2, and Erk1/2 and that this pathway is aberrant in individuals with XLP. PMID- 15814677 TI - Rapid functional exhaustion and deletion of CTL following immunization with recombinant adenovirus. AB - Replication-deficient adenoviruses (recombinant adenovirus (rec-AdV)) expressing different transgenes are widely used vectors for gene therapy and vaccination. In this study, we describe the tolerization of transgene-specific CTL following administration of beta-galactosidase (beta gal)-recombinant adenovirus (Ad-LacZ). Using MHC class I tetramers to track beta gal-specific CTL, we found that a significant expansion of beta gal-specific CTL was restricted to a very narrow dose range. Functional analysis revealed that adenovirus-induced beta gal specific CTL produced only very low amounts of effector cytokines and were unable to exhibit cytolytic activity in a 51Cr release assay. Furthermore, Ad-LacZ vaccination failed to efficiently clear established beta gal-positive tumors. The impaired function of Ad-LacZ-induced CTL correlated with the presence of persisting beta gal Ag in the liver. A further increase in the peripheral Ag load by injection of Ad-LacZ into SM-LacZ transgenic mice which express beta gal as self-Ag exclusively in peripheral nonlymphoid organs, resulted in the physical deletion of beta gal-specific CTL. Our results indicate first that CTL deletion in the course of adenoviral vaccination is preceded by their functional impairment and second, that the outcome of rec-AdV vaccination depends critically on the Ag load in peripheral tissues. PMID- 15814678 TI - Altered migration, recruitment, and somatic hypermutation in the early response of marginal zone B cells to T cell-dependent antigen. AB - The early responses of follicular (Fo) and marginal zone (MZ) B cells to T cell dependent Ag were compared using anti-hen egg lysozyme (HEL+) B cells capable of class switch recombination and somatic hypermutation (SHM). Purified CD21/(35int)CD23high Fo and CD21/35(high)CD23low MZ splenic B cells from SW(HEL) Ig-transgenic mice were transferred into wild-type recipients and challenged with HEL-sheep RBC. Responding HEL+ B cells from both populations switched efficiently to IgG1, generated syndecan-1+ Ab-secreting cells, and exhibited equivalent rates of proliferation. However, the expansion of HEL+ MZ B cells lagged significantly behind that of HEL+ Fo B cells due to less efficient homing to the outer periarteriolar lymphatic sheath and reduced recruitment into the proliferative response. Despite the equivalent rates of class switch recombination, the onset of SHM was delayed in the MZ subset, indicating that these two activation-induced cytidine deaminase-dependent events are uncoupled in the early response of MZ B cells. Migration of HEL+ B cells into germinal centers coincided with the onset of SHM, occurring more rapidly with Fo vs MZ responders. These results are consistent with the concept that Fo and MZ B cells have evolved to specialize in T cell-dependent and T-independent responses respectively. PMID- 15814679 TI - Suppressive oligodeoxynucleotides protect mice from lethal endotoxic shock. AB - Endotoxic shock is a life-threatening condition caused by exposure to bacterial LPS. LPS triggers the release of acute phase, proinflammatory, and Th1 cytokines that facilitate the development of endotoxic shock. Synthetic oligodeoxynucleotides (ODN) expressing suppressive TTAGGG motifs effectively down regulate the production of proinflammatory and Th1 cytokines elicited by a variety of immune stimuli. The current results demonstrate that suppressive ODN protect mice from LPS-induced endotoxic shock. Underlying this protective effect is the ability of suppressive ODN to bind to and prevent the phosphorylation of STAT1 and STAT4, thereby blocking the signaling cascade mediated by LPS-induced IFN-beta and IL-12. These findings suggest that suppressive ODN might be of use in the treatment of endotoxic shock. PMID- 15814680 TI - Microtubule disruption suppresses allergic response through the inhibition of calcium influx in the mast cell degranulation pathway. AB - Mast cells are secretory cells that release their granules, which contain inflammatory mediators. Some recent data suggested that cytoskeletons play a role in this process. However, the role of microtubules in Ca2+ signaling has not yet been well defined. In this study, we demonstrate that the microtubule cytoskeleton is important to maintain Ca2+ influx in the degranulation pathway of mast cells, using the microtubule depolymerizers nocodazole and colchicine. The microtubule depolymerizers inhibited Ag-induced degranulation in RBL-2H3 cells and bone marrow-derived mast cells. When the cells were stimulated with Ag in the presence of the microtubule depolymerizers, the Ca2+ influx was decreased without affecting Ca2+ release from the endoplasmic reticulum (ER). Capacitative Ca2+ entry, which was induced by inhibitors of Ca(2+)-ATPase in the ER membrane, thapsigargin and cyclopiazonic acid, was also decreased by nocodazole. Fluorescent probe analysis demonstrated that nocodazole disrupted microtubule formation and changed the cytoplasmic distribution of the ER. The microtubule depolymerizers attenuated the passive cutaneous anaphylaxis reaction in back skin of Sprague Dawley rats. These results suggest that the microtubule cytoskeleton in mast cells is important to maintain Ag-induced capacitative Ca2+ entry, which is responsible for degranulation and the allergic response. PMID- 15814681 TI - NK cells inhibit T cell proliferation via p21-mediated cell cycle arrest. AB - NK cells have been shown to influence immune responses via direct interaction with cells of the adaptive immune system, such as dendritic cells, B cells, and T cells. A role for NK cells in down-regulation of T cell responses has been implicated in several studies; however, the underlying mechanism of this suppression has remained elusive. In this study we show that dark Agouti rat NK cells inhibit syngeneic T cell proliferation via up-regulation of the cell cycle inhibitor, p21, resulting in a G0/G1 stage cell cycle arrest. The inhibition is cell-cell contact dependent, reversible, and Ag nonspecific. Interestingly, NK cells do not inhibit IL-2 secretion or IL-2R up-regulation and do not induce T cell death. Thus, our results show that NK cells do not affect early T cell activation events, but specifically inhibit T cell proliferation by direct interaction with T cells. Our findings suggest that NK cells may play an important role in maintaining immune homeostasis by directly regulating clonal expansion of activated T cells. This novel mechanism of T cell regulation by NK cells provides insight into NK cell-mediated regulation of adaptive immunity and provides a mechanistic link between NK cell function and suppression of T cell responses. PMID- 15814682 TI - Does the frequency and avidity spectrum of the neuroantigen-specific T cells in the blood mirror the autoimmune process in the central nervous system of mice undergoing experimental allergic encephalomyelitis? AB - In humans, studies of autoreactive T cells that mediate multiple sclerosis have been largely confined to testing peripheral blood lymphocytes. Little is known how such measurements reflect the disease-mediating autoreactive T cells in the CNS. This information is also not available for murine experimental allergic encephalomyelitis (EAE); the low number of T cells that can be obtained from the blood or the brain of mice prevented such comparisons. We used single-cell resolution IFN-gamma ELISPOT assays to measure the frequencies and functional avidities of myelin basic protein (MBP:87-99)-specific CD4 cells in SJL mice immunized with this peptide. Functional MBP:87-99-specific IFN-gamma-producing cells were present in the CNS during clinical signs of EAE, but not during phases of recovery. In contrast, MBP:87-99-specific T cells persisted in the blood during all stages of the disease, and were also present in mice that did not develop EAE. Therefore, the increased frequency of MBP:87-99-reactive T cells in the blood reliably reflected the primed state, but not the inflammatory activity of these cells in the brain. The functional avidity of the MBP:87-99-reactive T cells was identical in the brain and blood and did not change over 2 mo as the mice progressed from acute to chronic EAE. Therefore, high-affinity T cells did not become selectively enriched in the target organ, and avidity maturation of the MBP:87-99-specific T cell repertoire did not occur in the observation period. The data may help the interpretation of measurements made with peripheral blood lymphocytes of multiple sclerosis patients. PMID- 15814683 TI - Gamma delta T cell homeostasis is controlled by IL-7 and IL-15 together with subset-specific factors. AB - Among T cell subsets, gamma delta T cells uniquely display an Ag receptor-based tissue distribution, but what defines their preferential homing and homeostasis is unknown. To address this question, we studied the resources that control gamma delta T cell homeostasis in secondary lymphoid organs. We found that gamma delta and alpha beta T cells are controlled by partially overlapping resources, because acute homeostatic proliferation of gamma delta T cells was inhibited by an intact alpha beta T cell compartment, and both populations were dependent on IL-7 and IL 15. Significantly, to undergo acute homeostatic proliferation, gamma delta T cells also required their own depletion. Thus, gamma delta T cell homeostasis is maintained by trophic cytokines commonly used by other types of lymphoid cells, as well as by additional, as yet unidentified, gamma delta-specific factors. PMID- 15814684 TI - Rac2 regulates neutrophil chemotaxis, superoxide production, and myeloid colony formation through multiple distinct effector pathways. AB - Polymorphonuclear neutrophils (PMN) are an important component of the innate immune system. We have shown previously that migration and superoxide (O2*-) production, as well as some kinase signaling pathways are compromised in mice deficient in the Ras-related Rho GTPase Rac2. In this study, we demonstrate that Rac2 controls chemotaxis and superoxide production via distinct pathways and is critical for development of myeloid colonies in vitro. The Rac2 mutants V36A, F37A, and N39A all bind to both Pak1 and p67(phox), yet are unable to rescue superoxide production and chemotaxis when expressed in Rac2-/- PMN. In contrast, the N43A mutant, which binds to Por1 (Arfaptin 2), p67phox, and Pak1, is able to rescue superoxide production but not chemotaxis. The F37A mutant, demonstrated to have reduced binding to Por1, shows reduced rescue of fMLP-induced chemotaxis. Finally, the Rac2Y40C mutant that is defective in binding to all three potential downstream effectors (Pak1, p67phox, and Por1) is unable to rescue chemotaxis, motility, or superoxide production, but is able to rescue defective growth of myeloid colonies in vitro. These findings suggest that binding to any single effector is not sufficient to rescue the distinct cellular phenotypes of Rac2-/- PMN, implicating multiple, distinct, and potentially parallel effector pathways. PMID- 15814685 TI - Inhibitory NK receptor Ly49Q is expressed on subsets of dendritic cells in a cellular maturation- and cytokine stimulation-dependent manner. AB - Ly49Q is a member of the Ly49 family that is expressed on Gr-1+ cells but not on NK and NKT cells. Ly49Q appears to be involved in regulating cytoskeletal architectures through ITIM-mediated signaling. We provide evidence that dendritic cells (DCs) of certain maturational states expressed Ly49Q, and that IFN-alpha plays an important role in its regulation. Freshly prepared murine plasmacytoid pre-DCs as well as Flt3L-induced plasmacytoid pre-DCs expressed Ly49Q, whereas freshly prepared myeloid DCs did not. However, GM-CSF-induced myeloid DCs showed low levels of Ly49Q expression, and this was significantly enhanced by IFN-alpha. In contrast, other cytokines and ligands for TLRs such as TNF-alpha, IL-6, LPS, and CpG-ODN had little or no effect on Ly49Q expression. Plasmacytoid pre-DCs in all mouse strains examined expressed Ly49Q. Constitutive expression of Ly49Q on myeloid DCs was observed in three restricted mouse strains including 129, NZB, and NZW. As can be seen in other Ly49 family members, Ly49Q expression was affected by MHC class I expression. At the same time, Ly49Q possessed polymorphisms, including at least three alleles. The polymorphic residues lay within the stalk and carbohydrate recognition domain, and two of them, in loop 3 and loop 6 of the carbohydrate recognition domain, are located in the region implicated in the interaction of Ly49A with H-2D(d). Therefore, depending on IFN alpha, our results imply that Ly49Q serves a role for the biological functions of certain DC subsets through recognition of MHC class I or related molecules. PMID- 15814686 TI - Suppressive effect of IL-4 on IL-13-induced genes in mouse lung. AB - Although IL-4 signals through two receptors, IL-4R alpha/common gamma-chain (gamma(c)) and IL-4R alpha/IL-13R alpha1, and only the latter is also activated by IL-13, IL-13 contributes more than IL-4 to goblet cell hyperplasia and airway hyperresponsiveness in murine asthma. To determine whether unique gene induction by IL-13 might contribute to its greater proasthmatic effects, mice were inoculated intratracheally with IL-4 or IL-13, and pulmonary gene induction was compared by gene microarray and real-time PCR. Only the collagen alpha2 type VI (Ca2T6) gene and three small proline-rich protein (SPRR) genes were reproducibly induced > 4-fold more by IL-13 than by IL-4. Preferential IL-13 gene induction was not attributable to B cells, T cells, or differences in cytokine potency. IL 4 signaling through IL-4R alpha/gamma(c) suppresses Ca2T6 and SPRR gene expression in normal mice and induces these genes in RAG2/gamma(c)-deficient mice. Although IL-4, but not IL-13, induces IL-12 and IFN-gamma, which suppress many effects of IL-4, IL-12 suppresses only the Ca2T6 gene, and IL-4-induced IFN gamma production does not suppress the Ca2T6 or SPRR genes. Thus, IL-4 induces genes in addition to IL-12 that suppress STAT6-mediated SPRR gene induction. These results provide a potential explanation for the dominant role of IL-13 in induction of goblet cell hyperplasia and airway hyperresponsiveness in asthma. PMID- 15814687 TI - Signals from CD28 induce stable epigenetic modification of the IL-2 promoter. AB - CD28 costimulation controls multiple aspects of T cell function, including the expression of proinflammatory cytokine genes. One of these genes encodes IL-2, a growth factor that influences T cell proliferation, survival, and differentiation. Antigenic signaling in the absence of CD28 costimulation leads to anergy, a mechanism of tolerance that renders CD4+ T cells unable to produce IL-2. The molecular mechanisms by which CD28 costimulatory signals induce gene expression are not fully understood. In eukaryotic cells, the expression of many genes is influenced by their physical structure at the level of DNA methylation and local chromatin remodeling. To address whether these epigenetic mechanisms are operative during CD28-dependent gene expression in CD4+ T cells, we compared cytosine methylation and chromatin structure at the IL-2 locus in fully activated CD4+ effector T cells and CD4+ T cells rendered anergic by TCR ligation in the absence of CD28 costimulation. Costimulation through CD28 led to marked, stable histone acetylation and loss of cytosine methylation at the IL-2 promoter/enhancer. This was accompanied by extensive remodeling of the chromatin in this region to a structure highly accessible to DNA binding proteins. Conversely, TCR activation in the absence of CD28 costimulation was not sufficient to promote histone acetylation or cytosine demethylation, and the IL-2 promoter/enhancer in anergic cells remained completely inaccessible. These data suggest that CD28 may function through epigenetic mechanisms to promote CD4+ T cell responses. PMID- 15814688 TI - Translation from cryptic reading frames of DNA vaccines generates an extended repertoire of immunogenic, MHC class I-restricted epitopes. AB - To test whether simple expression units used in DNA vaccines can generate immunogenic, MHC class I-binding epitopes by translating other than the primary open reading frame (ORF), we constructed a vector (pCI/SX) that encodes the small hepatitis B surface Ag in the primary ORF, and a C-terminal fragment (residue 344 832) of the polymerase (Pol) in an alternative (out-of-frame) reading frame. pCI/SX efficiently primed multispecific, HLA-A2-restricted CD8+ T cell responses to epitopes of hepatitis B surface Ag and of Pol (Pol3, Pol(803-811)). Pol3 containing products generated from pCI/SX were detected only by T cell assays, but not by biochemical assays. Priming Pol-specific T cell responses to epitopes generated from alternative ORFs depended on promoter sequences that drive transcription in the DNA vaccine (human CMV-derived promoter sequences being more efficient than SV40-derived promoter sequences). Human CMV promoter-driven Pol constructs encoding different Pol fragments in primary or alternative reading frames elicited comparable levels of Pol3-specific T cell responses. We confirmed efficient T cell priming to epitopes from alternative ORFs by constructing DNA vaccines that encode an SV40-derived cT(1-272) protein fused either in frame or out of frame with an immunogenic OVA fragment (OVA(18-385)). Similar OVA-specific CD8+ T cell responses were primed by both alternative vaccine constructs. Hence, DNA vaccine-stimulated T cell responses to epitopes generated from alternative ORFs seem to be a regular event, although its biological role and risks are largely unexplored. PMID- 15814689 TI - Fetal expression of Fas ligand is necessary and sufficient for induction of CD8 T cell tolerance to the fetal antigen H-Y during pregnancy. AB - Interaction of Fas with Fas ligand (FasL) is known to play a role in peripheral tolerance mediated by clonal deletion of Ag-specific T cells. We have assessed the requirement for Fas/FasL interactions during induction of tolerance to the fetus. Using H-Y-specific TCR transgenic mice, we have previously demonstrated that exposure of maternal T cells to H-Y expressed by male fetuses results in deletion of 50% of H-Y-specific maternal T cells. The remaining H-Y-specific T cells were hyporesponsive to H-Y as assayed by decreased proliferative ability and CTL activity. To determine whether Fas/FasL interactions contribute to induction of maternal T cell tolerance, responsiveness to fetal H-Y was assessed in H-Y-specific TCR transgenic pregnant females that were deficient in functional Fas or FasL. Surprisingly, both deletion and nondeletion components of tolerance were abrogated in TCR transgenic H-Y-specific lpr (Fas-deficient) or gld (FasL deficient) pregnant females. Experiments further revealed that expression of FasL by the fetus, but not by the mother, is necessary and sufficient for both components of maternal T cell tolerance to fetal Ags. Fas interaction with fetal FasL is thus critical for both deletion and hyporesponsiveness of H-Y-reactive CD8+ T cells during pregnancy. PMID- 15814690 TI - T cell-mediated delay of spontaneous mammary tumor onset: increased efficacy with in vivo versus in vitro activation. AB - Peripheral tolerance to shared Ags expressed on both tumors and normal self tissues presents a major barrier to T cell-based immunotherapy as a treatment for cancer. To assess the activity of tumor-specific T cells against spontaneously arising carcinomas in the context of shared Ag expression, we developed a model system whereby an identified tumor Ag, tumor ERK (tERK), is expressed transgenically on both normal mammary tissue and spontaneous mammary carcinomas. Transfer of in vitro-activated, tERK-specific DUC18 T cells delayed spontaneous tumor development in tERK-expressing mice when T cells were given before the development of palpable carcinomas. However, antitumor activity mediated by in vitro-activated DUC18 T cells, as measured by responsiveness against a transplanted tERK-expressing fibrosarcoma challenge, was lost within days of transfer. This loss was due to expression of tERK as a self-Ag on normal tissues and was independent of the presence of mammary tumors. In contrast, transferred naive DUC18 T cells maintained a long-term protective function in tERK-expressing mice. Ten-fold fewer naive T cells activated in vivo were able to replicate the delay in spontaneous tumor development achieved by in vitro-activated T cells. These results are in contrast to our earlier studies using transplanted tumors alone, in which in vitro-activated DUC18 T cells were more efficacious than naive DUC18 T cells and highlight the need to perform tumor studies in the presence of tumor Ag expression on normal self-tissue. PMID- 15814692 TI - Gamma delta T cells regulate the extent and duration of inflammation in the central nervous system by a Fas ligand-dependent mechanism. AB - Gamma delta T cells have been shown to regulate immune responses associated with inflammation, but the mechanism of this regulation is largely unknown. Using the experimental autoimmune encephalomyelitis (EAE) model of the human CNS autoimmune disease multiple sclerosis, we demonstrate that gamma delta T cells are important regulators of CNS inflammation. This was shown using gamma delta T cell-deficient mice that were unable to recover from EAE. The chronic disease was accompanied by a prolonged presence of both macrophages and lymphocytes in the CNS. This extended inflammatory response was due to alterations in both cell proliferation and death. In mice lacking gamma delta T cells, proliferation of encephalitogenic T cells was 3-fold higher, and caspase activity, indicating apoptosis, was 2-fold lower compared with those in control mice recovering from EAE. gamma delta T cell deficient mice reconstituted with wild-type gamma delta T cells recovered from EAE and resolved inflammation in the CNS, whereas mice reconstituted with Fas ligand-dysfunctional gamma delta T cells did not. Thus, gamma delta T cells regulate both inflammation in the CNS and disease recovery via Fas/Fas ligand induced apoptosis of encephalitogenic T cells, and a quick resolution of inflammation in the CNS is essential to prevent permanent damage to the CNS resulting in chronic disease. PMID- 15814691 TI - Glucose availability regulates IFN-gamma production and p70S6 kinase activation in CD8+ effector T cells. AB - Differentiation of CD8+ T cells from the naive to the effector state is accompanied by changes in basal gene expression profiles that parallel the acquisition of effector functions. Among these are metabolism genes, and we now show that 2C TCR transgenic effector CD8+ T cells express higher levels of glycolytic enzymes and display greater glucose uptake, a higher glycolytic rate, and increased lactate production compared with naive cells. To determine whether glucose was required for effector T cell functions, we regulated glucose availability in vitro. Glucose deprivation strongly inhibited IFN-gamma gene expression, whereas IL-2 production was little affected. Inhibition correlated with diminished phosphorylation of p70S6 kinase and eIF4E binding protein 1 and a requirement for de novo protein synthesis, whereas other signaling pathways known to regulate IFN-gamma expression were unaffected. Together, our data reveal that optimal induction of IFN-gamma transcription is a glucose-dependent process, indicate that there are undefined factors that influence IFN-gamma expression, and have implications for regulation of the effector phase of CD8+ T cell responses in tissue microenvironments. PMID- 15814693 TI - IL-12-independent LIGHT signaling enhances MHC class II disparate CD4+ T cell alloproliferation, IFN-gamma responses, and intestinal graft-versus-host disease. AB - Inhibition of LIGHT (a cellular ligand for herpes virus entry mediator and lymphotoxin receptor)/herpes simplex virus entry mediator (HVEM) and LIGHT/lymphotoxin beta receptor (LT beta R) interactions decreases mortality in MHC class I and II disparate graft-vs-host disease (GVHD). The present studies assessed the effects of these interactions on the generation of CD4+ T cell alloresponses in MHC class II-disparate MLC and GVHD. An inhibitor protein of LIGHT and LT alpha beta2 (LT beta R-Ig) and an inhibitor protein of LIGHT (HVEM Ig) caused similar decreases in alloresponses of control B6 or B6.129S1 IL12rb2(tm1Jm) (B6.IL12R-/-) spleen cells (SpC) in a MHC class II-disparate MLC. GVHD-induced wasting disease in MHC class II-disparate recipients of B6 CD4+ SpC who received either the LT beta R-Ig-encoding adenovirus (LT beta R-Ig Adv; 13.1 +/- 10.9%; n = 10; p = 0.0004) or the HVEM-Ig-encoding adenovirus (HVEM-Ig Adv; 16.4 +/- 9.9%; n = 13; p = 0.0008) was significantly reduced compared with that in recipients of a control adenovirus (30.4 +/- 8.8%; n = 13). Furthermore, gut GVHD histologic scores of recipients of B6 CD4+ SpC who received the LT beta R-Ig Adv (0.8 +/- 0.8; n = 5; p = 0.0007) or the HVEM-Ig Adv (1.4 +/- 0.5; n = 5; p = 0.008) were reduced compared with scores of recipients of a control adenovirus (2.5 +/- 0.75; n = 11). In the intestine, both LT beta R-Ig Adv and HVEM-Ig Adv decreased CD4+ T cells (0.35 +/- 0.4 x 10(6) (n = 6) vs 0.36 +/- 0.02 x 10(6) (n = 9); p = 0.03 and p = 0.007) compared with control adenovirus (0.86 +/- 0.42 x 10(6); n = 9). LIGHT is critical for optimal CD4+ T cell alloresponses in MHC class II-disparate MLC and GVHD. PMID- 15814694 TI - Distinct roles of dendritic cells and B cells in Va14Ja18 natural T cell activation in vivo. AB - Va14Ja18 natural T (iNKT) cells are innate, immunoregulatory lymphocytes that recognize CD1d-restricted lipid Ags such as alpha-galactosylceramide (alpha GalCer). The immunoregulatory functions of iNKT cells are dependent upon either IFN-gamma or IL-4 production by these cells. We hypothesized that alpha GalCer presentation by different CD1d-positive cell types elicits distinct iNKT cell functions. In this study we report that dendritic cells (DC) play a critical role in alpha GalCer-mediated activation of iNKT cells and subsequent transactivation of NK cells. Remarkably, B lymphocytes suppress DC-mediated iNKT and NK cell activation. Nevertheless, alpha GalCer presentation by B cells elicits low IL-4 responses from iNKT cells. This finding is particularly interesting because we demonstrate that NOD DC are defective in eliciting iNKT cell function, but their B cells preferentially activate this T cell subset to secrete low levels of IL-4. Thus, the differential immune outcome based on the type of APC that displays glycolipid Ags in vivo has implications for the design of therapies that harness the immunoregulatory functions of iNKT cells. PMID- 15814695 TI - Macrophage colony-stimulating factor drives cord blood monocyte differentiation into IL-10(high)IL-12absent dendritic cells with tolerogenic potential. AB - Immature dendritic cells (DCs) induce tolerance and mature DCs induce inflammatory immune responses. However, the likelihood of maturation of immature DCs in vivo limits its potential application for suppression of unwanted immune reactions in vivo. The aim of this study was to generate DCs with anti inflammatory properties in both the immature and mature states. GM-CSF combined with IL-4 drives monocyte differentiation into DCs. As M-CSF is a critical cytokine in development of the monocytic lineage and its level is dramatically elevated in immunosuppressive conditions, we investigated whether M-CSF could replace GM-CSF and generate DCs with distinct functions from umbilical cord blood monocytes. Highly purified umbilical cord blood monocytes cultured with M-CSF and IL-4, in a GM-CSF-independent fashion, differentiated into IL-10(high)IL-12absent cells with a DC phenotype (termed M-DC). Single time stimulation with immature DCs (both M-DCs and DCs) derived from cord blood induced hyporesponsive and regulatory CD4+ T cells. In contrast to mature DCs, mature M-DCs induced decreased Th1 differentiation and proliferation of naive CD4+ T cells in both primary and secondary allogeneic MLR and showed tolerogenic potential. These results demonstrate an unrecognized role for M-CSF in alternative differentiation of monocytes into anti-inflammatory M-DCs and suggest that M-CSF-induced DCs may be of use for suppressing unwanted immune responses. PMID- 15814696 TI - T regulatory-1 cells induce IgG4 production by B cells: role of IL-10. AB - The study was aimed to find out whether T cells with a regulatory profile could regulate the secretion of IgG4. Using tetanus Ag we found that PBMC of healthy human donors responded to exogenous IL-10 by down-regulating IgG1 and increasing IgG4 secretion. IgE was not affected. To investigate the direct effect of IL-10 producing T cells on B cells, we generated T cell clones (TCC) with two different cytokine profiles: first, IL-10high, IL-2low, IL-4low TCC, and second, IL-10low, IL-2high, IL-4high. The T cell-dependent Ab secretion was measured by coculturing purified CD19+ B cells and the TCC. Interestingly, we found that IgG4 production in the coculture correlated with the TCC production of IL-10 (r2 = 0.352, p = 0.0001), but not with IL-2, IL-4, nor IFN-gamma. IgE showed only a trend with regard to IL-4. Further, there was decreased Ab secretion in the absence of T-B cell contact. IL-10 also induced IgG4 when added to a Th1 TCC-B cell coculture system. The present study thus shows that in T-B cell coculture, IL-10, if induced by the TCC or added to the system, down-regulates the immune response by inducing IgG4 secretion. This establishes a direct implication of IL-10 in humoral hyporesponsiveness, particularly in compartments where the T-B cell interplay determines the subsequent immune response. The correlation between IgG4 and IL-10 (r2 = 0.352) indicates that IL-10 is an important but not the only factor for IgG4 induction. PMID- 15814697 TI - Tumor vaccine based on cell surface expression of DcR3/TR6. AB - DcR3/TR6, a secreted protein belonging to the TNF receptor superfamily, interacts with lymphotoxin-like, exhibits inducible expression, and competes with herpes simplex virus glycoprotein D for herpes virus entrance mediator (LIGHT), Fas ligand (FasL), and TL1A, all members of the TNF superfamily. Solid-phase TR6 can trigger reverse signaling of LIGHT and FasL expressed on T cells, and lead to T cell costimulation. In this study, we engineered tumor cells to express cell surface TR6 and used these cells as a tumor vaccine. We demonstrated that mastocytoma P815 cells expressing surface TR6 (TR6-P815) effectively augmented the T cells response in vitro and ex vivo in terms of proliferation, as well as IL-2 and IFN-gamma secretion. TR6-P815 cells had reduced tumorigenicity compared with parental P815 cells. When inactivated TR6-P815 cells were employed as a vaccine, they protected the mice from challenge with live parental P815 cells, and eliminated established P815 tumors. The cell surface TR6-based tumor vaccine was also effective against low antigenicity tumors, such as B16 melanoma; co administration of bacillus Calmette-Guerin further enhanced the vaccine's efficacy. Thus, cell surface TR6 expression is a useful addition to our tumor vaccine arsenal. PMID- 15814698 TI - Pattern of Kv beta subunit expression in macrophages depends upon proliferation and the mode of activation. AB - Voltage-dependent potassium channels (Kv) in leukocytes are involved in the immune response. In bone marrow-derived macrophages (BMDM), proliferation and activation induce delayed rectifier K+ currents, generated by Kv1.3, via transcriptional, translational, and posttranslational controls. Furthermore, modulatory Kv beta subunits coassociate with Kv alpha subunits, increasing channel diversity and function. In this study we have identified Kv beta subunits in mouse BMDM, studied their regulation during proliferation and activation, and analyzed K+ current parameters influenced by these proteins. BMDM express all isoforms of Kv beta1 (Kv beta1.1, Kv beta1.2, and Kv beta1.3) and Kv beta2 (Kv beta2.1), but not Kv beta4, the alternatively spliced murine Kv beta3 variant. M CSF-dependent proliferation induced all Kv beta isoforms. However, LPS- and TNF alpha-induced activation differentially regulated these subunits. Although LPS increased Kv beta1.3, reduced Kv beta1.2, and maintained Kv beta1.1 mRNA levels constant, TNF-alpha up-regulated Kv beta1.1, down-regulated Kv beta1.2, and left Kv beta1.3 expression unchanged. Moreover, in contrast to TNF-alpha, M-CSF- and LPS- up-regulated Kv beta2.1. K+ currents from M-CSF- and LPS-stimulated BMDM exhibited faster inactivation, whereas TNF-alpha increased tau values. Although in M-CSF-stimulated cells the half-inactivation voltage shifted to more positive potentials, the incubation with LPS and TNF-alpha resulted in a hyperpolarizing displacement similar to that in resting BMDM. Furthermore, activation time constants of K+ currents and the kinetics of the tail currents were different depending upon the mode of activation. Our results indicate that differential Kv beta expression modifies the electrical properties of Kv in BMDM, dependent upon proliferation and the mode of activation. This could determine physiologically appropriate surface channel complexes, allowing for greater flexibility in the precise regulation of the immune response. PMID- 15814699 TI - A role for SATB1, a nuclear matrix association region-binding protein, in the development of CD8SP thymocytes and peripheral T lymphocytes. AB - Studies have suggested that binding of the SATB1 protein to L2a, a matrix association region located 4.5 kb 5' to the mouse CD8alpha gene, positively affects CD8 expression in T cells. Therefore, experiments were performed to determine the effect on T cell development of reduced expression of SATB1. Because homozygous SATB1-null mice do not survive to adulthood due to non-thymus autonomous defects, mice were produced that were homozygous for a T cell-specific SATB1-antisense transgene and heterozygous for a SATB1-null allele. Thymic SATB1 protein was reduced significantly in these mice, and the major cellular phenotype observed was a significant reduction in the percentage of CD8SP T cells in thymus, spleen, and lymph nodes. Mice were smaller than wild type but generally healthy, and besides a general reduction in cellularity and a slight increase in surface CD3 expression on CD8SP thymocytes, the composition of the thymus was similar to wild type. The reduction in thymic SATB1 does not lead to the variegated expression of CD8-negative single positive thymocytes seen upon deletion of several regulatory elements and suggested by others to reflect failure to activate the CD8 locus. Thus, the present results point to an essential role for SATB1 late in the development and maturation of CD8SP T cells. PMID- 15814700 TI - Dysfunction of simian immunodeficiency virus/simian human immunodeficiency virus induced IL-2 expression by central memory CD4+ T lymphocytes. AB - Production of IL-2 and IFN-gamma by CD4+ T lymphocytes is important for the maintenance of a functional immune system in infected individuals. In the present study, we assessed the cytokine production profiles of functionally distinct subsets of CD4+ T lymphocytes in rhesus monkeys infected with pathogenic or attenuated SIV/simian human immunodeficiency virus (SHIV) isolates, and these responses were compared with those in vaccinated monkeys that were protected from immunodeficiency following pathogenic SHIV challenge. We observed that preserved central memory CD4+ T lymphocyte production of SIV/SHIV-induced IL-2 was associated with disease protection following primate lentivirus infection. Persisting clinical protection in vaccinated and challenged monkeys is thus correlated with a preserved capacity of the peripheral blood central memory CD4+ T cells to express this important immunomodulatory cytokine. PMID- 15814701 TI - IL-6 increases primed cell expansion and survival. AB - Cytochrome c-specific CD4 T cells from transgenic donors transferred to syngeneic B10.A mice expand more vigorously upon immunization if exogenous IL-6 is provided during the initial phase of immunization. The resultant increase in the frequency and number of Ag-specific cells is observed in the blood, lymph nodes, spleen, liver, and lung and persists for at least 3 mo. Treatment of immunized recipients with anti-IL-6 or use of IL-6 knockout recipients reduced the frequency of Ag specific CD4 T cells during a comparable period, indicating that IL-6 is physiologically involved in the expansion of memory and/or effector cells and thus in the persistence of memory. IL-6 did not alter the duration of Ag presenting activity. Both CFSE dilution studies and labeling with BrdU indicated that IL-6 does not effect proliferative rates of responding CD4 T cells. By contrast, annexin V staining was diminished in responding cells from the IL-6 treated animals, particularly among those cells that had undergone five or more divisions. These results indicate that IL-6 reduces the level of apoptosis among Ag-stimulated cells; thus, it plays a central role in determining numbers of memory and/or effector CD4 T cells in response to immunization over extended periods. PMID- 15814702 TI - Recognition of human cytomegalovirus by human primary immunoglobulins identifies an innate foundation to an adaptive immune response. AB - Most primates, including humans, are chronically infected with cospecifically evolved, potentially pathogenic CMV. Abs that bind a 10-aa linear epitope (antigenic determinant 2 site 1) within the extracellular domain of human CMV glycoprotein B neutralize viral infectivity. In this study, we show that genes generated by recombinations involving two well-conserved human germline V elements (IGHV3-30 and IGKV3-11), and IGHJ4, encode primary Ig molecules that bind glycoprotein B at this key epitope. These particular V(H), J(H), and V(kappa) genes enable humans to generate through recombination and N nucleotide addition, a useful frequency of primary Igs that efficiently target this critical site on human CMV and thus confer an innate foundation for a specific adaptive response to this pathogen. PMID- 15814703 TI - HIV type 1 can act as an APC upon acquisition from the host cell of peptide loaded HLA-DR and CD86 molecules. AB - It is well documented that a wide range of host-derived cell surface constituents is inserted within HIV type 1 (HIV-1) and located on the exterior of the virion. Although no virus-associated protein of host origin has been shown to be absolutely required for virus replication, studies have revealed that many of these proteins are functional and can affect several steps of the virus life cycle. In this study, we found that HIV-1 acquires peptide-loaded class II MHC (MHC-II) and the costimulatory CD86 molecules from the host cell. Moreover, we present evidence that virions bearing such peptide-loaded MHC-II and CD86 proteins can lead to activation of the transcription factors NF-kappa B and NF-AT in an Ag-specific human T cell line. A linear correlation was found between activation of NF-kappa B and the amount of peptide-loaded MHC-II molecules inserted within HIV-1. Finally, transcription of unintegrated and integrated HIV 1 DNA was promoted upon exposure of peptide-specific human T cells to viruses bearing both peptide-loaded MHC-II and CD86 proteins. These data suggest that HIV 1 can operate as an APC depending on the nature of virus-anchored host cell membrane components. It can be proposed that HIV-1 can manipulate one of its primary targets through the process of incorporation of host-derived proteins. PMID- 15814704 TI - Functional requirements for signaling through the stimulatory and inhibitory mouse NKR-P1 (CD161) NK cell receptors. AB - The NK cell receptor protein 1 (NKR-P1) (CD161) molecules represent a family of type II transmembrane C-type lectin-like receptors expressed predominantly by NK cells. Despite sharing a common NK1.1 epitope, the mouse NKR-P1B and NKR-P1C receptors possess opposing functions in NK cell signaling. Engagement of NKR-P1C stimulates cytotoxicity of target cells, Ca2+ flux, phosphatidylinositol turnover, kinase activity, and cytokine production. In contrast, NKR-P1B engagement inhibits NK cell cytotoxicity. Nonetheless, it remains unclear how different signaling outcomes are mediated at the molecular level. Here, we demonstrate that both NKR-P1B and NKR-P1C associate with the tyrosine kinase, p56(lck). The interaction is mediated through the di-cysteine CxCP motif in the cytoplasmic domains of NKR-P1B/C. Disrupting this motif leads to abrogation of both stimulatory and inhibitory NKR-P1 signals. In addition, mutation of the consensus ITIM (LxYxxL) in NKR-P1B abolishes both its Src homology 2-containing protein tyrosine phosphatase-1 recruitment and inhibitory function. Strikingly, engagement of NKR-P1C on NK cells obtained from Lck-deficient mice failed to induce NK cytotoxicity. These results reveal a role for Lck in the initiation of NKR-P1 signals, and demonstrate a requirement for the ITIM in NKR-P1-mediated inhibition. PMID- 15814705 TI - B cell hyperresponsiveness and expansion of mature follicular B cells but not of marginal zone B cells in NFATc2/c3 double-deficient mice. AB - Marginal zone (MZ) B cells and peritoneal B-1 cells provide a first defense system of thymus-independent Ab responses against foreign pathogens and therefore share a number of functional properties. Recently, development of B-1a cells was shown to be controlled by the transcription factor NFATc1. We show here that mice deficient for NFATc2 and c3 display a distinct lower representation of MZ B cells, which is correlated with a reduced capturing of trinitrophenyl-Ficoll. In contrast, mature follicular B cells from NFATc2/c3-/- mice are strongly increased in number. NFATc2/c3-/- B cells exhibit a marked increase in BCR-induced intracellular Ca2+ mobilization and proliferation. However, trinitrophenyl-Ficoll specific IgM and IgG3 responses of NFATc2/c3-deficient mice are intact, and chimeric mice reconstituted with NFATc2/3-deficient B cells show a normal number of MZ B cells and normal BCR responses. These observations suggest that the strongly elevated Th2 cytokine milieu in NFATc2/c3-deficient mice leads to a hyperactivation of mature, follicular B cells, whereas MZ B cells are less responsive to these signals. PMID- 15814706 TI - Exocytosis of CTLA-4 is dependent on phospholipase D and ADP ribosylation factor 1 and stimulated during activation of regulatory T cells. AB - CTLA-4 is an essential protein in the regulation of T cell responses that interacts with two ligands found on the surface of APCs (CD80 and CD86). CTLA-4 is itself poorly expressed on the T cell surface and is predominantly localized to intracellular compartments. We have studied the mechanisms involved in the delivery of CTLA-4 to the cell surface using a model Chinese hamster ovary cell system and compared this with activated and regulatory human T cells. We have shown that expression of CTLA-4 at the plasma membrane (PM) is controlled by exocytosis of CTLA-4-containing vesicles and followed by rapid endocytosis. Using selective inhibitors and dominant negative mutants, we have shown that exocytosis of CTLA-4 is dependent on the activity of the GTPase ADP ribosylation factor-1 and on phospholipase D activity. CTLA-4 was identified in a perinuclear compartment overlapping with the cis-Golgi marker GM-130 but did not colocalize strongly with lysosomal markers such as CD63 and lysosome-associated membrane protein. In regulatory T cells, activation of phospholipase D was sufficient to trigger release of CTLA-4 to the PM but did not inhibit endocytosis. Taken together, these data suggest that CTLA-4 may be stored in a specialized compartment in regulatory T cells that can be triggered rapidly for deployment to the PM in a phospholipase D- and ADP ribosylation factor-1-dependent manner. PMID- 15814707 TI - Increased immunogenicity of an anchor-modified tumor-associated antigen is due to the enhanced stability of the peptide/MHC complex: implications for vaccine design. AB - The use of "anchor-fixed" altered peptide ligands is of considerable interest in the development of therapeutic vaccines for cancer and infectious diseases, but the mechanism by which successful altered peptide ligands elicit enhanced immunity is unclear. In this study, we have determined the crystallographic structure of a major tumor rejection Ag, gp100(209-217), in complex with the HLA A*0201 (HLA-A2) molecule, as well as the structure of a modified version of the peptide which substitutes methionine for threonine at position 2 (T2M; gp100(209 2M)). The T2M-modified peptide, which is more immunogenic in vitro and in vivo, binds HLA-A2 with a approximately 9-fold greater affinity and has a approximately 7-fold slower dissociation rate at physiological temperature. Within the limit of the crystallographic data, the T2M substitution does not alter the structure of the peptide/HLA-A2 complex. Consistent with this finding, in peripheral blood from 95 human subjects, we were unable to identify higher frequencies of T cells specific for either the native or modified peptide. These data strongly support the conclusion that the greater immunogenicity of the gp100(209-2M) peptide is due to the enhanced stability of the peptide/MHC complex, validating the anchor fixing approach for generating therapeutic vaccine candidates. Thermodynamic data suggest that the enhanced stability of the T2M-modified peptide/HLA-A2 complex is attributable to the increased hydrophobicity of the modified peptide, but the gain due to hydrophobicity is offset considerably by the loss of a hydrogen bond made by the native peptide to the HLA-A2 molecule. Our findings have broad implications for the optimization of current vaccine-design strategies. PMID- 15814708 TI - The enlarged population of marginal zone/CD1d(high) B lymphocytes in nonobese diabetic mice maps to diabetes susceptibility region Idd11. AB - The NOD mouse is an important experimental model for human type 1 diabetes. T cells are central to NOD pathogenesis, and their function in the autoimmune process of diabetes has been well studied. In contrast, although recognized as important players in disease induction, the role of B cells is not clearly understood. In this study we characterize different subpopulations of B cells and demonstrate that marginal zone (MZ) B cells are expanded 2- to 3-fold in NOD mice compared with nondiabetic C57BL/6 (B6) mice. The NOD MZ B cells displayed a normal surface marker profile and localized to the MZ region in the NOD spleen. Moreover, the MZ B cell population developed early during the ontogeny of NOD mice. By 3 wk of age, around the time when autoreactive T cells are first activated, a significant MZ B cell population of adult phenotype was found in NOD, but not B6, mice. Using an F2(B6 x NOD) cross in a genome-wide scan, we map the control of this trait to a region on chromosome 4 (logarithm of odds score, 4.4) which includes the Idd11 and Idd9 diabetes susceptibility loci, supporting the hypothesis that this B cell trait is related to the development of diabetes in the NOD mouse. PMID- 15814709 TI - Intraepithelial cell neutralization of HIV-1 replication by IgA. AB - HIV is transmitted sexually through mucosal surfaces where IgA Abs are the first line of immune defense. In this study, we used paired IgA and IgG mAbs against HIV gp160 to study intraepithelial cell neutralization and inhibition of HIV replication. African green monkey kidney cells, Vero C1008, polarizable epithelial cells transfected to express the polymeric Ig receptor (pIgR), were transfected with HIV proviral DNA, and intracellular neutralization mediated by the mAbs was assessed. D47A and D19A IgA, which neutralized HIV in a conventional assay, potently inhibited intracellular HIV replication as assessed by infecting HeLa-CD4-long terminal repeat/beta-galactosidase cells (human cervical carcinoma cell line) and CEMx174 cells (human T cell line) with apical supernatant, basolateral medium, and cell lysate from transfected cells. D47A also inhibited the production of virus as assessed by direct assay of p24. In contrast, D47 and D19 IgG, sharing the same V regions, but which were not transcytosed by the pIgR, did not inhibit intracellular HIV replication, nor did D47A and D19A IgA in pIgR- cells, incapable of transcytosing IgA. Confocal immunofluorescence microscopy showed prominent colocalization of HIV protein and D47A, in agreement with the intracellular neutralization data. D10A, which did not neutralize HIV in the conventional assay, and irrelevant IgA did not show intracellular neutralization or colocalization. Control studies with two kinds of conditioned medium confirmed that HIV neutralization had indeed occurred inside the cells. Thus, during its transcytosis through epithelial cells, HIV-specific IgA can neutralize HIV replication. PMID- 15814710 TI - Polymorphonuclear neutrophils improve replication of Chlamydia pneumoniae in vivo upon MyD88-dependent attraction. AB - Chlamydia pneumoniae, an obligate intracellular bacterium, causes pneumonia in humans and mice. In this study, we show that GR1+/CD45+ polymorphonuclear neutrophils (PMN) surprisingly increase the bacterial load of C. pneumoniae in vivo. Upon intranasal infection of wild-type mice, the lung weight is increased; the cytokines TNF, IL-12p40, and IFN-gamma, as well as the chemokines keratinocyte-derived chemokine, MCP-1, and MIP-2 are secreted; and GR1+/CD45+ PMN are recruited into lungs 3 days postinfection. In contrast, in infected MyD88 deficient mice, which lack a key adaptor molecule in the signaling cascade of TLRs and IL-1R family members, the increase of the lung weight is attenuated, and from the analyzed cyto- and chemokines, only IL-12p40 is detectable. Upon infection, almost no influx of inflammatory cells into lungs of MyD88-deficient mice can be observed. Six days postinfection, however, MyD88-deficient mice were able to produce TNF, IFN-gamma, keratinocyte-derived chemokine, and MCP-1 in amounts similar to wild-type mice, but failed to secrete IL-12p40 and MIP-2. At this time point, the infection increased the lung weight to a level similar to wild-type mice. Curiously, the chlamydial burden in MyD88-deficient mice 3 days postinfection is lower than in wild-type mice, a finding that can be reproduced in wild-type mice by depletion of GR1+ cells. In analyzing how PMN influence the chlamydial burden in vivo, we find that PMN are infected and enhance the replication of C. pneumoniae in epithelial cells. Thus, the lower chlamydial burden in MyD88-deficient mice can be explained by the failure to recruit PMN. PMID- 15814711 TI - Microbial antigen triggers rapid mobilization of TNF-alpha to the surface of mouse neutrophils transforming them into inducers of high-level dendritic cell TNF-alpha production. AB - Neutrophils play a critical role in early immunity to many microbial pathogens, and this may in part be due to their ability to release immunoregulatory cytokines and chemokines during infection. Here, we demonstrate by flow cytometric analysis that mouse polymorphonuclear leukocytes (PMN) up-regulate surface expression of TNF-alpha within 10 min of stimulation with LPS, and that this is followed by gradual loss over a period of 18 h. Early increases in surface TNF-alpha expression correlated with loss of intracellular pools of preformed TNF-alpha. Nevertheless, extended incubation with LPS resulted in increased levels of TNF-alpha mRNA synthesis and replenishment of intracellular cytokine. After triggering with LPS, PMN acquired the ability to induce dendritic cell (DC) TNF-alpha and IL-12 production. Transwell assays demonstrated that high level DC TNF-alpha production induced by LPS-triggered neutrophils was dependent upon cell-to-cell contact and neutrophil TNF-alpha, but neither was required for neutrophil instruction of DC IL-12 synthesis. The data suggest that microbial Ag triggered mouse PMN acquire the capacity to deliver potent DC-activating signals through elaboration of cytokines and direct interactions at the cell surface. PMID- 15814712 TI - Transmembrane TNF is sufficient to initiate cell migration and granuloma formation and provide acute, but not long-term, control of Mycobacterium tuberculosis infection. AB - TNF is critical for immunity against Mycobacterium tuberculosis infection; however, the relative contributions of the soluble and transmembrane forms of TNF in this immunity are unknown. Using memTNF mice, which express only the transmembrane form of TNF, we have addressed this question. Wild-type (WT), TNF-/ , and transmembrane TNF (memTNF) mice were infected with M. tuberculosis by aerosol. TNF-/- mice developed overwhelming infection with extensive pulmonary necrosis and died after only 33 days. memTNF mice, like WT mice, contained bacterial growth for over 16 wk, developed an Ag-specific T cell response, and initially displayed compact granulomas, comprised of both lymphocytes and macrophages. Expression of mRNA for the chemokines CXCL10, CCL3, CCL5, and CCL7 was comparable in both WT and memTNF mice. As the infection progressed, however, the pulmonary lesions in memTNF mice became larger and more diffuse, with increased neutrophil accumulation and necrosis. This was accompanied by increased influx of activated memory T cells into the lungs of memTNF mice. Eventually, these mice succumbed to infection with a mean time to death of 170 days. The expression of memTNF on T cells is functionally important because the transfer of T cells from memTNF, but not TNF-/- mice, into either RAG-/- or TNF-/- mice conferred the same survival advantage on the M. tuberculosis-infected recipient mice, as the transfer of WT T cells. Therefore, memTNF, in the absence of soluble TNF, is sufficient to control acute, but not chronic, M. tuberculosis infection, in part through its expression on T cells. PMID- 15814713 TI - Molecular basis for the potency of IL-10-deficient dendritic cells as a highly efficient APC system for activating Th1 response. AB - Identification and targeting of novel immunobiological factors that regulate the induction of Th1 cells are crucial for designing effective vaccines against certain intracellular pathogens, including Chlamydia. IL-10-deficient dendritic cells (DC) are potent APCs and effective cellular vaccines that activate a high frequency of specific Th1 cells. To elucidate the molecular basis for the potency of the IL-10-deficient APC system, we tested the hypothesis that Chlamydia Ag primed IL-10 knockout (IL-10KO) DC are quantitatively and qualitatively distinct in their metabolic characteristics relating to T cell activation. Using a combination of RT-PCR, two-dimensional gel electrophoresis, and MALDI-TOF-based proteomics analyses, the transcriptional and translational activities of Chlamydia-pulsed DC from wild-type and IL-10KO mice were assessed. IL-10 deficiency caused early maturation and activation of pulsed DC (i.e., high CD11c, CD40, CD80, CD83, CD86, IL-1, IL-12, and the T cell-attracting chemokine CCL27/CTACK) and consequently an enhanced ability to process and present Ags for a rapid and robust T cell activation. Supporting comparative proteomics revealed further that IL-10 deficient DC possess specific immunobiological properties, e.g., the T cell-attracting chemokine CCL27/CTACK, calcium-dependent protein kinase, and the IL-1/IL-12 inducer, NKR-P1A (CD161), which differentiated them immunologically from wild-type DC that express molecules relating to anti inflammatory, differentiative, and metabolic processes, e.g., the anti-IL-12 molecule peroxisome proliferator-activated receptor-alpha and thymidine kinase. Collectively, these results provide a molecular basis for the high Th1-activating capacity of IL-10KO APC and may provide unique immunomodulation targets when designing vaccines against pathogens controlled by T cell immunity. PMID- 15814714 TI - Differential regulation of beta-defensin expression in human skin by microbial stimuli. AB - In response to infection, epithelia mount an innate immune response that includes the production of antimicrobial peptides. However, the pathways that connect infection and inflammation with the induction of antimicrobial peptides in epithelia are not understood. We analyzed the molecular links between infection and the expression of three antimicrobial peptides of the beta-defensin family, human beta-defensin (hBD)-1, hBD-2, and hBD-3 in the human epidermis. After exposure to microbe-derived molecules, both monocytes and lymphocytes stimulated the epidermal expression of hBD-1, hBD-2, and hBD-3. The induced expression of hBD-3 was mediated by transactivation of the epidermal growth factor receptor. The mechanisms of induction of hBD-1 and hBD-3 were distinct from each other and from the IL-1-dependent induction of hBD-2 expression. Thus during inflammation, epidermal expression of beta-defensins is mediated by at least three different mechanisms. PMID- 15814715 TI - STAT1 signaling regulates tumor-associated macrophage-mediated T cell deletion. AB - It is well established that tumor progression is associated with the accumulation of myeloid suppressive cells, which in mice include Gr-1+ immature myeloid cells and F4/80+ macrophages. The paradox is that with the exception of terminal stages of the disease or chemotherapy treatment, tumor-bearing mice or cancer patients do not display a profound systemic immune suppression. We therefore raised the question as to whether myeloid cell-mediated T cell suppression is controlled at a local level at the site of the tumor. We have demonstrated that after adoptive transfer to tumor-bearing recipients, Gr-1+ (immature myeloid cells) freshly isolated from spleens of tumor-bearing mice become F4/80+ tumor-associated macrophages (TAM). These TAM, but not F4/80+ macrophages or Gr-1+ cells freshly isolated from spleens of tumor-bearing or naive mice were able to inhibit T cell mediated immune response in vitro via induction of T cell apoptosis. Arginase and NO were both responsible for the apoptotic mechanism, and were seen only in TAM, but not in freshly isolated Gr1+ cells. Using the analysis of STAT activity in combination with STAT knockout mice, we have determined that STAT1, but not STAT3 or STAT6, was responsible for TAM-suppressive activity. PMID- 15814716 TI - Roles for CXC chemokine ligands 10 and 11 in recruiting CD4+ T cells to HIV-1 infected monocyte-derived macrophages, dendritic cells, and lymph nodes. AB - We investigated roles for chemoattractants in dissemination of HIV-1 by examining the induction of T cell-active chemokines in HIV-1-infected human monocyte derived macrophages and dendritic cells. Of the 12 chemokines analyzed, mRNAs for two, CXCL10 and CXCL11, ligands for the chemokine receptor CXCR3, were up regulated in both cell types upon infection by HIV-1. Induction of these chemokine genes in infected cultures was dependent on both viral entry and reverse transcriptase activity, but not on the HIV-1 envelope glycoprotein. Conditioned medium from infected cells was chemotactic for freshly isolated human CD4+ T cells, and chemotaxis was abolished by pretreatment with an Ab against CXCR3. A lymph node from an HIV-1-infected individual expressed CXCL10 and CXCL11 mRNAs in the paracortex, including venules, as detected by in situ hybridization, whereas neither mRNA was detected after highly active antiretroviral therapy. Because CCR5 on CD4+ T cells is found predominantly on cells that also express CXCR3, these data implicate CXCL10 and CXCL11 in the recruitment of susceptible T cells to HIV-1-infected lymph nodes, macrophages, and dendritic cells. This recruitment might enhance the sequestration of T cells in infected lymphoid organs and the spread of infection between cells, contributing to the immunopathology of AIDS. PMID- 15814717 TI - Cathelicidin mediates innate intestinal defense against colonization with epithelial adherent bacterial pathogens. AB - Cathelicidin-related antimicrobial peptide (mCRAMP), the sole murine cathelicidin, is encoded by the gene Cnlp. We show that mCRAMP expression in the intestinal tract is largely restricted to surface epithelial cells in the colon. Synthetic mCRAMP had antimicrobial activity against the murine enteric pathogen Citrobacter rodentium, which like the related clinically important human pathogens enteropathogenic Escherichia coli and enterohemorrhagic E. coli, adheres to the apical membrane of intestinal epithelial cells. Colon epithelial cell extracts from Cnlp+/+ mice had significantly greater antimicrobial activity against C. rodentium than those of mutant Cnlp-/- mice that lack mCRAMP. Cnlp-/- mice developed significantly greater colon surface and crypt epithelial cell colonization, surface epithelial cell damage, and systemic dissemination of infection than Cnlp+/+ mice after oral infection with C. rodentium. Moreover, Cnlp+/+ mice were protected from oral infections with C. rodentium inocula that infected the majority of Cnlp-/- mice. These results establish cathelicidin as an important component of innate antimicrobial defense in the colon. PMID- 15814718 TI - Receptor-binding domain of severe acute respiratory syndrome coronavirus spike protein contains multiple conformation-dependent epitopes that induce highly potent neutralizing antibodies. AB - The spike (S) protein of severe acute respiratory syndrome associated coronavirus (SARS-CoV) is a major antigenic determinant capable of inducing protective immunity. Recently, a small fragment on the SARS-CoV S protein (residues 318-510) was characterized as a minimal receptor-binding domain (RBD), which mediates virus binding to angiotensin-converting enzyme 2, the functional receptor on susceptible cells. In this study, we demonstrated that a fusion protein containing RBD linked to human IgG1 Fc fragment (designated RBD-Fc) induced high titer of RBD-specific Abs in the immunized mice. The mouse antisera effectively neutralized infection by both SARS-CoV and SARS pseudovirus with mean 50% neutralization titers of 1/15,360 and 1/24,737, respectively. The neutralization determinants on the RBD of S protein were characterized by a panel of 27 mAbs isolated from the immunized mice. Six groups of conformation-dependent epitopes, designated as Conf I-VI, and two adjacent linear epitopes were identified by ELISA and binding competition assays. The Conf IV and Conf V mAbs significantly blocked RBD-Fc binding to angiotensin-converting enzyme 2, suggesting that their epitopes overlap with the receptor-binding sites in the S protein. Most of the mAbs (23 of 25) that recognized the conformational epitopes possessed potent neutralizing activities against SARS pseudovirus with 50% neutralizing dose ranging from 0.005 to 6.569 microg/ml. Therefore, the RBD of SARS S protein contains multiple conformational epitopes capable of inducing potent neutralizing Ab responses, and is an important target site for developing vaccines and immunotherapeutics. PMID- 15814719 TI - Prevention of relapse after chemotherapy in a chronic intracellular infection: mechanisms in experimental visceral leishmaniasis. AB - In visceral leishmaniasis, chemotherapy probably seldom eradicates all parasites in tissue macrophages; nevertheless, most T cell-intact patients show long lasting clinical cure after treatment despite residual intracellular infection. To characterize prevention of posttreatment relapse, amphotericin B was used to kill approximately 90-95% of Leishmania donovani in livers of mice deficient in mechanisms of acquired antileishmanial resistance. Recrudescence subsequently developed 1) in animals deficient in both CD4 and CD8 T cells as well as CD40L mediated T cell costimulation, but not in a) CD4 or CD8 cells alone, b) NK cell lytic activity, or c) ICAM-1-recruited monocytes; and 2) in mice deficient in IFN gamma, but not in the IFN-gamma-inducing cytokines, a) IL-12, b) IL-12 and IL-23, or c) IL-18. Posttreatment recrudescence also did not develop in animals deficient in macrophage phagocyte NADPH oxidase (phox) or inducible NO synthase (iNOS) alone or, surprisingly, in those deficient in both phox and iNOS. Therefore, regulation of the intracellular replication of residual Leishmania donovani that escape chemotherapy evolves to a host mechanism distinguishable from initial acquired resistance at the T cell, cytokine, and macrophage levels. Posttreatment, either CD8 or CD4 cells can direct the response, IL-12 is not required, and iNOS and phox, the activated macrophage's primary IFN-gamma inducible leishmanicidal pathways, both become dispensable. PMID- 15814720 TI - Removal of regulatory T cell activity reverses hyporesponsiveness and leads to filarial parasite clearance in vivo. AB - Human filarial parasites cause chronic infection associated with long-term down regulation of the host's immune response. We show here that CD4+ T cell regulation is the main determinant of parasite survival. In a laboratory model of infection, using Litomosoides sigmodontis in BALB/c mice, parasites establish for >60 days in the thoracic cavity. During infection, CD4+ T cells at this site express increasing levels of CD25, CTLA-4, and glucocorticoid-induced TNF receptor family-related gene (GITR), and by day 60, up to 70% are CTLA 4(+)GITR(high), with a lesser fraction coexpressing CD25. Upon Ag stimulation, CD4(+)CTLA-4(+)GITR(high) cells are hyporesponsive for proliferation and cytokine production. To test the hypothesis that regulatory T cell activity maintains hyporesponsiveness and prolongs infection, we treated mice with Abs to CD25 and GITR. Combined Ab treatment was able to overcome an established infection, resulting in a 73% reduction in parasite numbers (p < 0.01). Parasite killing was accompanied by increased Ag-specific immune responses and markedly reduced levels of CTLA-4 expression. The action of the CD25(+)GITR+ cells was IL-10 independent as in vivo neutralization of IL-10R did not restore the ability of the immune system to kill parasites. These data suggest that regulatory T cells act, in an IL-10-independent manner, to suppress host immunity to filariasis. PMID- 15814721 TI - Anthrax edema toxin cooperates with lethal toxin to impair cytokine secretion during infection of dendritic cells. AB - Bacillus anthracis secretes two critical virulence factors, lethal toxin (LT) and edema toxin (ET). In this study, we show that murine bone marrow-derived dendritic cells (DC) infected with B. anthracis strains secreting ET exhibit a very different cytokine secretion pattern than DC infected with B. anthracis strains secreting LT, both toxins, or a nontoxinogenic strain. ET produced during infection selectively inhibits the production of IL-12p70 and TNF-alpha, whereas LT targets IL-10 and TNF-alpha production. To confirm the direct role of the toxins, we show that purified ET and LT similarly disrupt cytokine secretion by DC infected with a nontoxinogenic strain. These effects can be reversed by specific inhibitors of each toxin. Furthermore, ET inhibits in vivo IL-12p70 and IFN-gamma secretion induced by LPS. These results suggest that ET produced during infection impairs DC functions and cooperates with LT to suppress the innate immune response. This may represent a new strategy developed by B. anthracis to escape the host immune response. PMID- 15814722 TI - Apoptosis induced by the toll-like receptor adaptor TRIF is dependent on its receptor interacting protein homotypic interaction motif. AB - TLRs detect specific molecular features of microorganisms and subsequently engage distinct signaling networks through the differential use of Toll/IL-1R (TIR) domain-containing adapter proteins. In this study, we investigated the control of apoptosis by the TIR domain-containing adapter proteins MyD88, TIR-domain containing adapter protein (TIRAP), TIR-domain-containing adapter-inducing IFN beta (TRIF), TRIF-related adapter molecule (TRAM), and sterile alpha motifs and beta-catenin/armadillo repeats (SARM). Upon overexpression, TRIF was the sole TIR adapter to potently engage mammalian cell death signaling pathways. TRIF-induced cell death required caspase activity initiated by the Fas/Apo-1-associated DD protein-caspase-8 axis and was unaffected by inhibitors of the intrinsic apoptotic machinery. The proapoptotic potential of TRIF mapped to the C-terminal region that was found to harbor a receptor interacting protein (RIP) homotypic interaction motif (RHIM). TRIF physically interacted with the RHIM-containing proteins RIP1 and RIP3, and deletion and mutational analyses revealed that the RHIM in TRIF was essential for TRIF-induced apoptosis and contributed to TRIF induced NF-kappa B activation. The domain that was required for induction of apoptosis could activate NF-kappa B but not IFN regulatory factor-3, yet the activation of NF-kappa B could be blocked by superrepressor I kappa B alpha without blocking apoptosis. Thus, the ability of TRIF to induce apoptosis was not dependent on its ability to activate either IFN regulatory factor-3 or NF-kappa B but was dependent on the presence of an intact RHIM. TRIF serves as an adaptor for both TLR3 and TLR4, receptors that are activated by dsRNA and LPS, respectively. These molecular motifs are encountered during viral and bacterial infection, and the apoptosis that occurs when TRIF is engaged represents an important host defense to limit the spread of infection. PMID- 15814723 TI - Surfactant protein-D regulates soluble CD14 through matrix metalloproteinase-12. AB - Surfactant protein D (SP-D) and CD14 are important innate immune defense molecules that mediate clearance of pathogens and apoptotic cells from the lung. To test whether CD14 expression and function were influenced by SP-D, the surface expression of CD14 was assessed on alveolar macrophages from SP-D-/- mice. CD14 was reduced on alveolar macrophages from SP-D-/- mice and was associated with reduced uptake of LPS and decreased production of TNF-alpha after LPS stimulation. CD14 is proteolytically cleaved from the cell surface to form a soluble peptide. Soluble CD14 (sCD14) was increased in the bronchoalveolar lavage fluid from SP-D-/- mice. Because matrix metalloproteinase (MMP)-9 and -12 activities were increased in the lungs of SP-D-/- mice, the role of these metalloproteases in the production of sCD14 was assessed. sCD14 was decreased in both MMP(9-/-)/SP-D-/- and MMP12(-/-)/SP-D-/- mice demonstrating MMP-9 and MMP-12 contribute to proteolytic shedding of CD14. The increased sCD14 seen in SP-D-/- mice was dependent upon the activation of MMP-12 via an MMP-9-dependent mechanism. Supporting this observation, MMP-12 caused the release of sCD14 from RAW 264.7 cells in vitro. In conclusion, SP-D influences innate host defense, in part, by regulating sCD14 in a process mediated by MMP-9 and MMP-12. PMID- 15814724 TI - Activation-induced depletion of protein kinase C alpha provokes desensitization of monocytes/macrophages in sepsis. AB - Sepsis accounts for the majority of fatal casualties in critically ill patients, because extensive research failed to significantly improve appropriate therapy strategies. Thus, understanding molecular mechanisms initiating the septic phenotype is important. Symptoms of septic disease are often associated with monocyte/macrophage desensitization. In this study, we provide evidence that a desensitized cellular phenotype is characterized by an attenuated oxidative burst. Inhibition of the oxidative burst and depletion of protein kinase C alpha (PKC alpha) were correlated in septic patients. To prove that PKC alpha down regulation indeed attenuated the oxidative burst, we set up a cell culture model to mimic desensitized monocytes/macrophages. We show that LPS/IFN-gamma-treatment of RAW264.7 and U937 cells lowered PKC alpha expression and went on to confirm these data in primary human monocyte-derived macrophages. To establish a role of PKC alpha in cellular desensitization, we overexpressed PKC alpha in RAW264.7 and U937 cells and tested for phorbolester-elicited superoxide formation following LPS/IFN-gamma-pretreatment. Inhibition of the oxidative burst, i.e., cellular desensitization, was clearly reversed in cells overexpressing PKC alpha, pointing to PKC alpha as the major transmitter in eliciting the oxidative burst in monocytes/macrophages. However, PKC alpha inactivation by transfecting a catalytically inactive PKC alpha mutant attenuated superoxide formation. We suggest that depletion of PKC alpha in monocytes from septic patients contributes to cellular desensitization, giving rise to clinical symptoms of sepsis. PMID- 15814725 TI - Anthrax lethal toxin blocks MAPK kinase-dependent IL-2 production in CD4+ T cells. AB - Anthrax lethal toxin (LT) is a critical virulence factor that cleaves and inactivates MAPK kinases (MAPKKs) in host cells and has been proposed as a therapeutic target in the treatment of human anthrax infections. Despite the potential use of anti-toxin agents in humans, the standard activity assays for anthrax LT are currently based on cytotoxic actions of anthrax LT that are cell-, strain-, and species-specific, which have not been demonstrated to occur in human cells. We now report that T cell proliferation and IL-2 production inversely correlate with anthrax LT levels in human cell assays. The model CD4+ T cell tumor line, Jurkat, is a susceptible target for the specific protease action of anthrax LT. Anthrax LT cleaves and inactivates MAPKKs in Jurkat cells, whereas not affecting proximal or parallel TCR signal transduction pathways. Moreover, anthrax LT specifically inhibits PMA/ionomycin- and anti-CD3-induced IL-2 production in Jurkat cells. An inhibitor of the protease activity of anthrax LT completely restores IL-2 production by anthrax LT-treated Jurkat cells. Anthrax LT acts on primary CD4+ T cells as well, cleaving MAPKKs and leading to a 95% reduction in anti-CD3-induced proliferation and IL-2 production. These findings not only will be useful in the development of new human cell-based bioassays for the activity of anthrax LT, but they also suggest new mechanisms that facilitate immune evasion by Bacillus anthracis. Specifically, anthrax LT inhibits IL-2 production and proliferative responses in CD4+ T cells, thereby blocking functions that are pivotal in the regulation of immune responses. PMID- 15814726 TI - Early establishment of gamma-herpesvirus latency: implications for immune control. AB - The human gamma-herpesviruses, EBV and Kaposi's sarcoma-associated herpesvirus, infect >90% of the population worldwide, and latent infection is associated with numerous malignancies. Rational vaccination and therapeutic strategies require an understanding of virus-host interactions during the initial asymptomatic infection. Primary EBV infection is associated with virus replication at epithelial sites and entry into the circulating B lymphocyte pool. The virus exploits the life cycle of the B cell and latency is maintained long term in resting memory B cells. In this study, using a murine gamma-herpesvirus model, we demonstrate an early dominance of latent virus at the site of infection, with lung B cells harboring virus almost immediately after infection. These data reinforce the central role of the B cell not only in the later phase of infection, but early in the initial infection. Early inhibition of lytic replication does not impact the progression of the latent infection, and latency is established in lymphoid tissues following infection with a replication deficient mutant virus. These data demonstrate that lytic viral replication is not a requirement for gamma-herpesvirus latency in vivo and suggest that viral latency can be disseminated by cellular proliferation. These observations emphasize that prophylactic vaccination strategies must target latent gamma herpesvirus at the site of infection. PMID- 15814727 TI - Leukotriene B4 receptor-1 is essential for allergen-mediated recruitment of CD8+ T cells and airway hyperresponsiveness. AB - Recent studies in both human and rodents have indicated that in addition to CD4+ T cells, CD8+ T cells play an important role in allergic inflammation. We previously demonstrated that allergen-sensitized and -challenged CD8-deficient (CD8-/-) mice develop significantly lower airway hyperresponsiveness (AHR), eosinophilic inflammation, and IL-13 levels in bronchoalveolar lavage fluid compared with wild-type mice, and that all these responses were restored by adoptive transfer of in vivo-primed CD8+ T cells or in vitro-generated effector CD8+ T cells (T(EFF)). Recently, leukotriene B4 and its high affinity receptor, BLT1, have been shown to mediate in vitro-generated T(EFF) recruitment into inflamed tissues. In this study we investigated whether BLT1 is essential for the development of CD8+ T cell-mediated allergic AHR and inflammation. Adoptive transfer of in vivo-primed BLT1+/+, but not BLT1-/-, CD8+ T cells into sensitized and challenged CD8-/- mice restored AHR, eosinophilic inflammation, and IL-13 levels. Moreover, when adoptively transferred into sensitized CD8-/- mice, in vitro-generated BLT1+/+, but not BLT1-/-, T(EFF) accumulated in the lung and mediated these altered airway responses to allergen challenge. These data are the first to show both a functional and an essential role for BLT1 in allergen mediated CD8+ T(EFF) recruitment into the lung and development of AHR and airway inflammation. PMID- 15814728 TI - Dominant role for TL1A/DR3 pathway in IL-12 plus IL-18-induced IFN-gamma production by peripheral blood and mucosal CCR9+ T lymphocytes. AB - The TNF-like cytokine TL1A augments IFN-gamma production by anti-CD3 plus anti CD28 and IL-12/IL-18-stimulated peripheral blood (PB) T cells. However, only a small subset of PB T cells respond to TL1A stimulation with IFN-gamma production. PB CCR9+ T cells represent a small subset of circulating T cells with mucosal T cell characteristics and a Th1/Tr1 cytokine profile. In the current study, we show that TL1A enhanced IFN-gamma production by TCR- or CD2/CD28-stimulated CCR9(+)CD4+ PB T cells. However, TL1A had the most pronounced effect on augmenting IFN-gamma production by IL-12/IL-18-primed CCR9(+)CD4+ PB T cells. TL1A enhanced both the percentage and the mean fluorescence intensity of IFN gamma in CCR9(+)CD4+ T cells as assessed by intracellular cytokine staining. IL 12 plus IL-18 up-regulated DR3 expression in CCR9(+)CD4+ T cells but had negligible effect on CCR9(-)CD4+ T cells. CCR9(+)CD4+ T cells isolated from the small intestine showed a 37- to 105-fold enhancement of IFN-gamma production when TL1A was added to the IL-12/IL18 cytokine combination. Cell membrane-expressed TL1A was preferentially expressed in CCR9(+)CD4+ PB T cells, and a blocking anti TL1A mAb inhibited IFN-gamma production by cytokine-primed CCR9(+)CD4+ T cells by approximately 50%. Our data show that the TL1A/DR3 pathway plays a dominant role in the ultimate level of cytokine-induced IFN-gamma production by CCR9+ mucosal and gut-homing PB T cells and could play an important role in Th1-mediated intestinal diseases, such as Crohn's disease, where increased expression of IL 12, IL-18, TL1A, and DR3 converge in the inflamed intestinal mucosa. PMID- 15814729 TI - IL-1beta-mediated innate immunity is amplified in the db/db mouse model of type 2 diabetes. AB - Chronic inflammation appears to play a critical role in type 2 diabetes and its complications. Here we tested the hypothesis that this inflammatory dysregulation affects the IL-1beta system and has functional consequences in the brain. Diabetic, db/db, and nondiabetic, db/+, mice were administered i.p. LPS, a potent cytokine inducer, at a dose of 100 microg/kg/mouse. db/db mouse innate immune associated sickness behavior was 14.8, 33, 44.7, and 34% greater than that of db/+ mice at 2, 4, 8, and 12 h, respectively. When a fixed dose of LPS was used (5 microg/mouse), db/db mouse sickness was again enhanced 18.4, 22.2, and 14.5% at 4, 8, and 12 h as compared with db/+ mice. In diabetic mice, peritoneal macrophages produced more IL-1beta in response to LPS, and peritoneal levels of IL-1beta induced by LPS were increased. Importantly, IL-1R antagonist and type 2 IL-1 receptor (IL-1R2) failed to up-regulate in response to LPS in db/db mice. Finally, both peripheral and central administration of IL-1beta, itself, induced sickness in db/db mice that mimicked the effects of peripheral LPS and was significantly greater than that seen in db/+ mice. Taken together, these results indicate that IL-1beta-mediated innate immunity is augmented in db/db mice both at the periphery and in the brain, and the mechanism is due to diabetes associated loss of IL-1beta counterregulation. PMID- 15814730 TI - Composition of the lectin pathway of complement in Gallus gallus: absence of mannan-binding lectin-associated serine protease-1 in birds. AB - The lectin pathway of complement is activated by multimolecular complexes that recognize and bind to microbial polysaccharides. These complexes comprise a multimeric carbohydrate recognition subunit (either mannan-binding lectin (MBL) or a ficolin), three MBL-associated serine proteases (MASP-1, -2, and -3), and MAp19 (a truncated product of the MASP-2 gene). In this study we report the cloning of chicken MASP-2, MASP-3, and MAp19 and the organization of their genes and those for chicken MBL and a novel ficolin. Mammals usually possess two MBL genes and two or three ficolin genes, but chickens have only one of each, both of which represent the undiversified ancestors of the mammalian genes. The primary structure of chicken MASP-2 is 54% identical with those of the human and mouse MASP-2, and the organization of its gene is the same as in mammals. MASP-3 is even more conserved; chicken MASP-3 shares approximately 75% of its residues with human and Xenopus MASP-3. It is more widely expressed than other lectin pathway components, suggesting a possible function of MASP-3 different from those of the other components. In mammals, MASP-1 and MASP-3 are alternatively spliced products of a single structural gene. We demonstrate the absence of MASP-1 in birds, possibly caused by the loss of MASP-1-specific exons during phylogeny. Despite the lack of MASP-1-like enzymatic activity in sera of chicken and other birds, avian lectin pathway complexes efficiently activate C4. PMID- 15814731 TI - In vivo activity of released cell wall lipids of Mycobacterium bovis bacillus Calmette-Guerin is due principally to trehalose mycolates. AB - The hallmark of Mycobacterium-induced pathology is granulomatous inflammation at the site of infection. Mycobacterial lipids are potent immunomodulators that contribute to the granulomatous response and are released in appreciable quantities by intracellular bacilli. Previously we investigated the granulomagenic nature of the peripheral cell wall lipids of Mycobacterium bovis bacillus Calmette-Guerin (BCG) by coating the lipids onto 90-microm diameter microspheres that were mixed into Matrigel matrix with syngeneic bone marrow derived macrophages and injected i.p. into mice. These studies demonstrated that BCG lipids elicit proinflammatory cytokines and recruit leukocytes. In the current study we determined the lipids responsible for this proinflammatory effect. BCG-derived cell wall lipids were fractionated and purified by liquid chromatography and preparative TLC. The isolated fractions including phosphatidylinositol dimannosides, cardiolipin, phosphatidylglycerol, phosphatidylethanolamine, trehalose monomycolate, trehalose dimycolate, and mycoside B. Trehalose dimycolate, when delivered to bone marrow-derived murine macrophages, induced the greatest secretion of IL-1beta, IL-6, and TNF-alpha in vitro. Trehalose dimycolate similarly induced the greatest secretion of these proinflammatory cytokines in ex vivo matrices over the course of 12 days. Trehalose monomycolate and dimycolate also induced profound neutrophil recruitment in vivo. Experiments with TLR2 or TLR4 gene-deficient mice revealed no defects in responses to trehalose mycolates, although MyD88-deficient mice manifested significantly reduced cell recruitment and cytokine production. These results demonstrate that the trehalose mycolates, particularly trehalose dimycolate, are the most bioactive lipids in the BCG extract, inducing a proinflammatory cascade that influences granuloma formation. PMID- 15814732 TI - TLR2 signaling in chondrocytes drives calcium pyrophosphate dihydrate and monosodium urate crystal-induced nitric oxide generation. AB - Microcrystals of calcium pyrophosphate dihydrate (CPPD) and monosodium urate (MSU) deposited in synovium and articular cartilage initiate joint inflammation and cartilage degradation in large part by binding and directly activating resident cells. TLRs trigger innate host defense responses to infectious pathogens, and the expression of certain TLRs by synovial fibroblasts has revealed the potential for innate immune responses to be triggered by mesenchymally derived resident cells in the joint. In this study we tested the hypothesis that chondrocytes also express TLRs and that one or more TLRs centrally mediate chondrocyte responsiveness to CPPD and MSU crystals in vitro. We detected TLR2 expression in normal articular chondrocytes and up-regulation of TLR2 in osteoarthritic cartilage chondrocytes in situ. We demonstrated that transient transfection of TLR2 signaling-negative regulator Toll-interacting protein or treatment with TLR2-blocking Ab suppressed CPPD and MSU crystal induced chondrocyte release of NO, an inflammatory mediator that promotes cartilage degeneration. Conversely, gain-of-function of TLR2 in normal chondrocytes via transfection was associated with increased CPPD and MSU crystal induced NO release. Canonical TLR signaling by parallel pathways involving MyD88, IL-1R-associated kinase 1, TNF receptor-associated factor 6, and IkappaB kinase and Rac1, PI3K, and Akt critically mediated NO release in chondrocytes stimulated by both CPPD and MSU crystals. We conclude that CPPD and MSU crystals critically use TLR2-mediated signaling in chondrocytes to trigger NO generation. Our results indicate the potential for innate immunity at the level of the articular chondrocyte to directly contribute to inflammatory and degenerative tissue reactions associated with both gout and pseudogout. PMID- 15814733 TI - Allergen drives class switching to IgE in the nasal mucosa in allergic rhinitis. AB - IgE-expressing B cells are over 1000 times more frequent in the nasal B cell than the peripheral blood B cell population. We have investigated the provenance of these B cells in the nasal mucosa in allergic rhinitis. It is generally accepted that expression of activation-induced cytidine deaminase and class switch recombination (CSR) occur in lymphoid tissue, implying that IgE-committed B cells must migrate through the circulation to the nasal mucosa. Our detection of mRNA for activation-induced cytidine, multiple germline gene transcripts, and epsilon circle transcripts in the nasal mucosa of allergic, in contrast to nonallergic control subjects, however, indicates that local CSR occurs in allergic rhinitis. The germline gene transcripts and epsilon circle transcripts in grass pollen allergic subjects are up-regulated during the season and also when biopsies from allergic subjects are incubated with the allergen ex vivo. These results demonstrate that allergen stimulates local CSR to IgE, revealing a potential target for topical therapies in allergic rhinitis. PMID- 15814734 TI - Cyclooxygenase 2 plays a pivotal role in the resolution of acute lung injury. AB - Acute lung injury (ALI) is a severe illness with excess mortality and no specific therapy. In its early exudative phase, neutrophil activation and accumulation in the lung lead to hypoxemia, widespread tissue damage, and respiratory failure. In clinical trials, inhibition of proinflammatory mediators has not proven effective. In this study, we pursued a new investigative strategy that emphasizes mediators promoting resolution from lung injury. A new spontaneously resolving experimental murine model of ALI from acid aspiration was developed to identify endogenous proresolving mechanisms. ALI increased cyclooxygenase 2 (COX-2) expression in murine lung. Selective pharmacologic inhibition or gene disruption of COX-2 blocked resolution of ALI. COX-2-derived products increased levels of the proresolving lipid mediators lipoxin A4 (LXA4) and, in the presence of aspirin, 15-epi-LXA4. Both LXA4 and 15-epi-LXA4 interact with the LXA4 receptor (ALX) to mediate anti-inflammatory actions. ALX expression was markedly induced by acid injury and transgenic mice with increased ALX expression displayed dramatic protection from ALI. Together, these findings indicate a protective role in ALI for COX-2-derived mediators, in part via enhanced lipoxin signaling, and carry potential therapeutic implications for this devastating clinical disorder. PMID- 15814735 TI - A2A adenosine receptors on bone marrow-derived cells protect liver from ischemia reperfusion injury. AB - Activation of the A2A adenosine receptor (A(2A)R) during reperfusion of various tissues has been found to markedly reduce ischemia-reperfusion injury. In this study, we used bone marrow transplantation (BMT) to create chimeric mice that either selectively lack or selectively express the A(2A)R on bone marrow-derived cells. Bolus i.p. injection of the selective A2A agonist, 4-[3-[6-amino-9-(5 cyclopropylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2 ynyl]-piperidine-1-carboxylic acid methyl ester (ATL313; 3 microg/kg), at the time of reperfusion protects wild-type (wt) mice from liver ischemia-reperfusion injury. ATL313 also protects wt/wt (donor/recipient BMT mouse chimera) and wt/knockout chimera but produces modest protection of knockout/wt chimera as assessed by alanine aminotransferase activity, induction of cytokine transcripts (RANTES, IFN-gamma-inducible protein-10, IL-1alpha, IL-1-beta, IL-1Ralpha, IL-18, IL-6, and IFN-gamma), or histological criteria. ATL313, which is highly selective for the A(2A)R, produces more liver protection of chimeric BMT mice than 4-[3-[6 amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl] prop-2-ynyl]-cyclohexanecarboxylic acid methyl ester, which is rapidly metabolized in mice to produce 4-[3-[6-amino-9-(5-ethylcarbamoyl-3,4-dihydroxy tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl]-cyclohexanecarboxylic acid, which has similar affinity for the A(2A)R and the proinflammatory A3 adenosine receptor. GFP chimera mice were created to show that vascular endothelial cells in the injured liver do not account for liver protection because they are not derived by transdifferentiation of bone marrow precursors. The data suggest that activation of the A(2A)R on bone marrow-derived cells is primarily responsible for protecting the liver from reperfusion injury. PMID- 15814736 TI - ERK1/2 regulates epidermal chemokine expression and skin inflammation. AB - Resident cell populations of the skin contribute to the inflammatory response by producing an array of chemokines, which attract leukocytes from the circulation. TNF-alpha is a major inducer of proinflammatory mediators in keratinocytes. We have recently observed that epidermal growth factor receptor (EGFR) signaling affects TNF-alpha-driven chemokine expression in epidermal keratinocytes, and its functional impairment increases the levels of crucial chemoattractants such as CCL2/MCP-1, CCL5/RANTES, and CXCL10/IFN-gamma-inducible protein-10. In this study, we report evidence that EGFR-dependent ERK1/2 activity is implicated in this mechanism. Abrogation of ERK1/2 activity with specific inhibitors increased chemokine expression in keratinocytes by enhancing mRNA stabilization. In mouse models, inflammatory response to irritants and T cell-mediated contact hypersensitivity were both aggravated when elicited in a skin area previously treated with an EGFR or a MAPK kinase 1/2 inhibitor. In contrast, impairment of p38alpha beta MAPK phosphorylation markedly attenuated these responses. Our data indicate that EGFR-dependent ERK1/2 activity in keratinocytes takes part to a homeostatic mechanism regulating inflammatory responses, and emphasize the distinct role of MAPKs as potential targets for manipulating inflammation in the skin. PMID- 15814737 TI - Fibrin deimination in synovial tissue is not specific for rheumatoid arthritis but commonly occurs during synovitides. AB - Autoantibodies to deiminated (citrullinated) proteins are the most specific serological markers of rheumatoid arthritis (RA). Deimination is critical in generating the peptidic epitopes they recognize. In the synovial tissue (ST), deiminated forms of the alpha- and beta-chains of fibrin are their major autoantigenic targets (anti-human fibrin(ogen) autoantibodies (AhFibA)). We investigated whether the presence of deiminated fibrin in the ST was specific for RA, because this could explain why AhFibA are RA specific. In 13 patients with RA and 19 patients with various other rheumatological disorders, knee ST biopsies were collected in macroscopically inflamed areas identified under arthroscopy. Synovitis was histopathologically confirmed in all of the biopsies. By immunoblotting, using antisera to fibrin, Abs to citrullyl residues, and AhFibA purified from RA sera, deiminated fibrin was evidenced in ST extracts from all of the patients. Moreover, variations in the degree of fibrin deimination were observed that were not related to the disease. Immunohistochemical analysis, using Abs to citrullyl residues and an antiserum to fibrin on adjacent serial sections of ST, confirmed the results because deiminated proteins colocalized with fibrin in RA as well as in control patients. Therefore, fibrin deimination in the ST is a general phenomenon associated to any synovitis, which does not necessarily induce a B autoimmune response with production of AhFibA. PMID- 15814738 TI - Autoimmune alterations induced by the New Zealand Black Lbw2 locus in BWF1 mice. AB - The New Zealand Black (NZB) Lbw2 locus (lupus NZB x New Zealand White (NZW) 2 locus) was previously linked to mortality and glomerulonephritis, but not to IgG autoantibodies, suggesting that it played a role in a later disease stage. To define its contribution, (NZB x NZW)F1 hybrids (BWF1) containing two, one, or no copies of this locus were generated. Lack of the NZB Lbw2 indeed reduced mortality and glomerulonephritis, but not serum levels of total and anti-DNA IgG Abs. There were, however, significant reductions in the B cell response to LPS, total and anti-DNA IgM and IgG Ab-forming cells, IgM Ab levels, and glomerular Ig deposits. Furthermore, although serum IgG autoantibody levels correlated poorly with kidney IgG deposits, the number of spontaneous IgG Ab-forming cells had a significant correlation. Genome-wide mapping of IgM anti-chromatin levels identified only Lbw2, and analysis of subinterval congenics tentatively reduced Lbw2 to approximately 5 Mb. Because no known genes associated with B cell activation and lupus are in this interval, Lbw2 probably represents a novel B cell activation gene. These findings establish the importance of Lbw2 in the BWF1 hybrid and indicate that Lbw2, by enhancing B cell hyperactivity, promotes the early polyclonal activation of B cells and subsequent production of autoantibodies. PMID- 15814739 TI - CCL1-CCR8 interactions: an axis mediating the recruitment of T cells and Langerhans-type dendritic cells to sites of atopic skin inflammation. AB - Atopic dermatitis represents a chronically relapsing skin disease with a steadily increasing prevalence of 10-20% in children. Skin-infiltrating T cells, dendritic cells (DC), and mast cells are thought to play a crucial role in its pathogenesis. We report that the expression of the CC chemokine CCL1 (I-309) is significantly and selectively up-regulated in atopic dermatitis in comparison to psoriasis, cutaneous lupus erythematosus, or normal skin. CCL1 serum levels of atopic dermatitis patients are significantly higher than levels in healthy individuals. DC, mast cells, and dermal endothelial cells are abundant sources of CCL1 during atopic skin inflammation and allergen challenge, and Staphylococcus aureus-derived products induce its production. In vitro, binding and cross linking of IgE on mast cells resulted in a significant up-regulation of this inflammatory chemokine. Its specific receptor, CCR8, is expressed on a small subset of circulating T cells and is abundantly expressed on interstitial DC, Langerhans cells generated in vitro, and their monocytic precursors. Although DC maintain their CCR8+ status during maturation, brief activation of circulating T cells recruits CCR8 from intracytoplamic stores to the cell surface. Moreover, the inflammatory and atopy-associated chemokine CCL1 synergizes with the homeostatic chemokine CXCL12 (SDF-1alpha) resulting in the recruitment of T cell and Langerhans cell-like DC. Taken together, these findings suggest that the axis CCL1-CCR8 links adaptive and innate immune functions that play a role in the initiation and amplification of atopic skin inflammation. PMID- 15814740 TI - CD4+ T cell responses to SSX-4 in melanoma patients. AB - Genes of the synovial sarcoma X breakpoint (SSX) family are expressed in different human tumors, including melanomas, but not in adult somatic tissues. Because of their specific expression at the tumor site, SSX-encoded Ags are potential targets for anticancer immunotherapy. In this study, we have analyzed CD4+ T cell responses directed against the Ag encoded by SSX-4. Upon in vitro stimulation of PBMC from four melanoma patients bearing Ag-expressing tumors with a pool of long peptides spanning the protein sequence, we detected and isolated SSX-4-specific CD4+ T cells recognizing several distinct antigenic sequences, mostly restricted by frequently expressed HLA class II alleles. The majority of the identified sequences were located within the Kruppel-associated box domain in the N-terminal region of the protein, indicating a high potential immunogenicity of this region. Together our data document the existence of CD4+ T cells specific for multiple SSX-4 derived sequences in circulating lymphocytes from melanoma patients and encourage further studies to assess the impact of SSX-4-specific T cell responses on disease evolution in cancer patients. PMID- 15814741 TI - Naive CD4+ T cells from lupus-prone Fas-intact MRL mice display TCR-mediated hyperproliferation due to intrinsic threshold defects in activation. AB - Autoreactive T cell activation is a consistent feature of murine lupus; however, the mechanism of such activation remains unclear. We hypothesized that naive CD4+ T cells in lupus have a lower threshold of activation through their TCR-CD3 complex that renders them more susceptible to stimulation with self-Ags. To test this hypothesis, we compared proliferation, IL-2 production, and single cell calcium signaling of naive CD4+ T cells isolated from Fas-intact MRL/+(Fas-lpr) mice with H-2k-matched B10.BR and CBA/CaJ controls, following anti-CD3 stimulation in the presence or absence of anti-CD28. We also assessed the responsiveness of naive CD4+ T cells isolated from Fas-intact MRL and control mice bearing a rearranged TCR specific for amino acids 88-104 of pigeon cytochrome c to cognate and low affinity peptide Ags presented by bone marrow matured dendritic cells. TCR transgenic and wild-type CD4+ T cells from MRL mice displayed a lower threshold of activation than control cells, a response that was class II MHC dependent. The rise in intracellular calcium in MRL vs controls was enhanced and prolonged following anti-CD3 triggering, suggestive of proximal defects in TCR-engendered signaling as the mechanism for the observed hyperactivity. These findings were observed as early as 1-2 mo postweaning and, based on analysis of F1 T cells, appeared to be dominantly expressed. This genetically altered threshold for activation of MRL T cells, a consequence of a proximal defect in CD3-mediated signal transduction, may contribute to the abrogation of T cell tolerance to self-Ags in lupus. PMID- 15814742 TI - Accelerated lymphocyte death in sepsis occurs by both the death receptor and mitochondrial pathways. AB - Patients with sepsis are immune compromised, as evidenced by their failure to clear their primary infection and their propensity to develop secondary infections with pathogens that are often not particularly virulent in normal healthy individuals. A potential mechanism for immunosuppression in sepsis is lymphocyte apoptosis, which may occur by either a death receptor or a mitochondrial-mediated pathway. A prospective study of blood samples from 71 patients with sepsis, 55 nonseptic patients, and 6 healthy volunteers was undertaken to quantitate lymphocyte apoptosis and determine cell death pathways and mechanisms of apoptosis. Apoptosis was evaluated by flow cytometry and Western blotting. Lymphocyte apoptosis was increased in CD4 and CD8 T cells, B cells (CD20), and NK cells (CD56) in septic vs nonseptic patients. Samples taken sequentially from 10 patients with sepsis showed that the degree of CD3 T cell apoptosis correlated with the activity of his/her sepsis. In septic patients, apoptotic lymphocytes were positive for active caspases 8 and 9, consistent with death occurring by both mitochondrial-mediated and receptor-mediated pathways. In support of the concept that both death pathways were operative, lymphocyte apoptosis occurred in cells with markedly decreased Bcl-2 (an inhibitor of mitochondrial-mediated apoptosis) as well as cells with normal concentrations of Bcl-2. In conclusion, apoptosis occurs in a broad range of lymphocyte subsets in patients with sepsis and correlates with the activity of the disease. Lymphocyte loss occurs by both death receptor and mitochondrial-mediated apoptosis, suggesting that there may be multiple triggers for lymphocyte apoptosis. PMID- 15814743 TI - Coinfection with Schistosoma mansoni is associated with decreased HIV-specific cytolysis and increased IL-10 production. AB - Impaired virus-specific immune responses have previously been observed with Schistosoma mansoni coinfection. We characterized Gag-specific responses in HIV-1 positive Ugandans with and without S. mansoni coinfection. We observed no significant difference in the frequency of IFN-gamma CD8+ T cells between the two groups. Interestingly, expression of CD107, a marker for cytolytic activity, was significantly lower in volunteers with S. mansoni coinfection compared with those with HIV-1 infection alone (p = 0.002). In contrast, the frequency of IL-10 positive Gag-specific CD8+ T cell responses was higher in volunteers with S. mansoni coinfection (p = 0.004). Analysis of human CMV-specific CD8+ T cell responses in the same individuals failed to reveal a similar pattern of altered CD107 and IL-10 expression. Our results suggest that S. mansoni coinfection is associated with decreased Gag-specific CD8+ cytolytic T cell responses and increased number of Gag-specific IL-10 positive CD8+ T cells. Our findings may have important implications toward the implementation of HIV preventive and therapeutic programs in Africa. PMID- 15814744 TI - Neutrophils that infiltrate the central nervous system regulate T cell responses. AB - Regulation of inflammatory responses is critical to progression of organ-specific autoimmune disease. Although many candidate cell types have been identified, immunoregulatory activity has rarely been directly assayed and never from the CNS. We have analyzed the regulatory capability of Gr-1high neutrophils isolated from the CNS of mice with experimental autoimmune encephalomyelitis. Proportions of neutrophils were markedly increased in the CNS of IFN-gamma-deficient mice. Strikingly, CNS-derived neutrophils, whether or not they derived from IFN-gamma deficient mice, were potent suppressors of T cell responses to myelin or adjuvant Ags. Neutrophil suppressor activity was absolutely dependent on IFN-gamma production by target T cells, and suppression was abrogated by blocking NO synthase. These data identify an immunoregulatory capacity for neutrophils, and indicate that interplay between IFN-gamma, NO, and activated Gr-1high neutrophils within the target organ determines the outcome of inflammatory and potentially autoimmune T cell responses. PMID- 15814746 TI - The health(y) cost of erythropoietin in orthopedic surgery. PMID- 15814747 TI - Clinical freedom's final refuge. PMID- 15814748 TI - Noncardiac surgery in patients with coronary artery stent: what should the anesthesiologist know? PMID- 15814749 TI - Erythropoietin is an effective clinical modality for reducing RBC transfusion in joint surgery. AB - PURPOSE: Efficacy trials of preoperative erythropoietin therapy (PET) recommend a dosing schedule that cannot always be adhered to in everyday clinical practice. Consequently, we instituted a flexible dosing schedule and routinely offered it to anemic patients [hemoglobin (Hb)] < or = 130 g x L(-1)) undergoing total joint arthroplasty (TJA). The purpose of this observational, cohort study was to assess the effectiveness of this practice in reducing red blood cell (RBC) transfusion. METHODS: After obtaining Institutional Ethics Board approval, data were collected prospectively on all patients who underwent TJA at our institution from July 1999 to June 2003. Patients with baseline Hb < or = 130 g x L(-1) were offered PET as follows: one to three sc injections (20,000 IU for those < or = 70 kg, and 40,000 IU for those > 70 kg) every three to seven days before surgery. Since treatment was not randomly assigned, multivariable logistic regression analysis and propensity score case-control matching were used to adjust for baseline differences in patient demographics and perioperative risk factors for RBC transfusion. The adjusted relationship between PET and RBC transfusion was then determined. RESULTS: Of the 1,782 patients that underwent TJA during the study period, 770 (47.9%) had a Hb < 130 g x L(-1). Of these patients, 214 received PET and their RBC transfusion rate was 16.4%, whereas the transfusion rate was 56.1% in those who did not receive PET (P < 0.0001). The adjusted odds ratio of RBC transfusion with PET was 0.33 (95% confidence interval = 0.21-0.49). CONCLUSION: PET, used as part of routine clinical practice, is an effective blood conservation modality. PMID- 15814750 TI - Use of hemoglobin raffimer for postoperative life-threatening anemia in a Jehovah's Witness. AB - PURPOSE: To describe the successful treatment of acute, life-threatening anemia with the oxygen therapeutic agent, hemoglobin (Hb) raffimer. CLINICAL FEATURES: A 53-yr-old female Jehovah's Witness developed severe anemia following total hip replacement. Due to prior patient directive, red blood cells were not transfused. Tachycardia, hypotension, electrocardiographic abnormalities and mental status changes developed with a nadir Hb concentration of 3.2 g x dL(-1). Hb raffimer is a purified, cross-linked, human Hb solution developed as a substitute for red blood cell Hb. After obtaining informed consent as well as Food and Drug Administration and Institutional Review Board approval for compassionate use, 2 L of Hb raffimer (Hemolink, Hemosol, Inc., Toronto, ON, Canada) were administered along with ferrous sulfate and epoetin alfa therapy. The patient's Hb level rose to 5.5 g x dL(-1) with resolution of symptoms. To allow recovery of red blood cell mass while maintaining Hb level > 4.5 g x dL(-1), additional 1000 mL doses of Hb raffimer were administered on postoperative days three, five and seven (total dose = 500 g Hb). The patient developed no serious adverse events related to treatment with Hb raffimer. By postoperative day 14, the patient's Hb level increased to 6.5 g x dL(-1) with a hematocrit of 23%. The patient was discharged. CONCLUSIONS: Use of Hb raffimer as a bridge to recovery of this patient's red blood cell mass may have prevented adverse clinical outcome. Because this product is a purified Hb solution devoid of other cellular components, it may be accepted as therapy by patients who, due to religious conviction, refuse allogeneic red blood cell transfusion. PMID- 15814751 TI - Changes in hematocrit based on incremental blood sampling: mathematical models perform poorly. AB - PURPOSE: Excessive blood sampling, with its inherent risks, is of growing concern among clinicians. We performed this study to measure the changes in hematocrit (Hct) during a laboratory investigation where multiple blood samples are collected. The performance of a simple mathematical model, used in clinical practice to predict Hct changes, is evaluated. METHODS: Eight healthy male volunteers participated in this study. The equation Hct(f) = Hct(i)*(EBV-BL)/EBV is used to predict changes in Hct. Where Hct(f) and Hct(i) are, respectively, the final and initial Hct, EBV is the estimated blood volume and BL is the blood loss. RESULTS: Thirty-five pharmacokinetic samples per subject were collected totalling 314 mL of BL. The Hct decreased from 44.2% +/- 2.2% to 39.9% +/- 2.5% (P = 0.001). On average, model predictions tended to have a discrete tendency to underestimate the Hct changes (-0.5% points of bias). While the predictions of the Hct were very accurate in 50% of the subjects, the discrepancy of the Hct predictions was clinically significant in the other 50% of the subjects. CONCLUSION: Consistent with the model prediction, this study demonstrated a significant reduction in the Hct values in healthy subjects undergoing incremental phlebotomy. On average, the model successfully predicted the decrease in Hct. However, the inter- and intra-individual variabilities in the Hct changes are clinically significant. In clinical settings, which are not well controlled environments, the variability is likely to be greater and the clinical use of the model cannot replace the need to monitor the Hct. PMID- 15814752 TI - The Stealth Station Image Guidance System may interfere with pulse oximetry. AB - PURPOSE: Interference on pulse oximetry can come from many sources. We found an additional source of interference from the Stealth Station. This article gives an overview of sources of pulse oximeter interference so that clinicians can better prevent them. Technical features: This article discusses the infrared disturbances caused by the Stealth Station. The Stealth Station is a frameless stereotactic positioning system that utilizes a three dimensional location system to measure the position of the patient and the surgical tools, and to relate those positions to previously recorded imaging. To understand the disturbance caused by the Stealth Station, we discuss its operation and that of pulse oximeter monitors. Pulse oximeter interference can come from volume artifacts, electrical and light noise, and can be caused by issues related to the patient. Because the passive Stealth Station contains a strong infrared light source, interference caused by light is a likely reason for the interference we noted. Pulse oximeters rely on the time-variant light signal modulated by arterial volume variations in the finger. Although relatively immune to static light sources, pulse oximeters are extremely sensitive to time-varying light sources. The light emitted by the passive Stealth Station is time-varying at 4 Hz and this is causing the pulse oximeter to provide invalid results. Shielding can generally be used to stop the light from the Stealth Station from being picked up by the pulse oximeter sensor. CONCLUSION: Infrared light interference can be very common, but is easily dealt with if one is aware of it. PMID- 15814753 TI - Survey of chronic pain practice by anesthesiologists in Canada. AB - PURPOSE: To describe the pattern of chronic pain practice (CPP) among anesthesiologists in Canada. METHODS: Following hospital Ethics Committee approval, a detailed postal questionnaire was sent to all active members of the Canadian Anesthesiologists' Society. A second mailing was conducted two months later. RESULTS: The overall response rate was 53%. While 38% of responding anesthesiologists were involved in CPP, in the majority of cases, this accounted for less than 20% of their clinical time. Thirty percent of those involved in CPP had previous training in pain management. The types of CPP included nerve blocks (84%) and pharmacological treatment (60%) in non-cancer pain (85%) and cancer pain (50%) patients. Ten percent and 28% of anesthesiologists were involved in research and teaching respectively while 26% were affiliated with a multidisciplinary clinic. The healthcare professions that anesthesiologists had access to or were directly working with in their practice were as follows: acupuncture (18%), nursing (36%), psychology (28%), psychiatry (35%) and physiotherapy (58%). Epidural steroid injection was the most commonly practiced intervention (82%). This was followed by trigger point injection (70%), stellate ganglion block (61%), occipital nerve block (60%) and lumbar sympathetic block (50%). Practice of interventional procedures was highly diverse. Seventy percent of anesthesiologists prescribed opioids as part of their CPP. However, half of them never incorporated an opioid agreement with patients. Opioids were most commonly used in the sustained release form. CONCLUSION: Approximately one-third of anesthesiologists surveyed incorporate chronic pain in their practice and their pattern of practice is widely diversified. PMID- 15814754 TI - Duration of analgesia is similar when 15, 20, 25 and 30 mL of ropivacaine 0.5% are administered via a femoral catheter. AB - PURPOSE: This dose-response study was designed to determine the most appropriate dose of ropivacaine 0.5% injected via an indwelling femoral catheter for perioperative peripheral analgesia for total knee replacement (TKR). METHODS: 84 patients were allocated randomly to four groups and received, via a femoral catheter, either 15, 20, 25 or 30 mL of ropivacaine 0.5% in a double-blind fashion. An anterior sciatic block with 20 mL bupivacaine 0.5% was also performed. The evolution of sensory block of femoral, obturator and lateral femoral cutaneous nerves and motor block of femoral nerve were tested every five minutes during the first 30 min. The percentage of patients with complete sensory block of both femoral and obturator nerves determined success rate. General anesthesia was then induced. After surgery, patient-controlled analgesia (PCA) with ropivacaine 0.2% was available via the femoral catheter. The interval between the initial injection and the first PCA administration determined duration of action. RESULTS: The duration of action was not different between the four solutions tested i.e., 534 +/- 379 min for 15 mL, 799 +/- 364 min for 20 mL, 624 +/- 342 min for 25 mL and 644 +/- 266 min for 30 mL. The percentage of patients with complete sensory femoral and obturator blocks was, respectively, 60%, 95%, 85% and 70% for 15, 20, 25 and 30 mL (P = 0.008/15 mL vs 20 mL). CONCLUSION: Although there is no difference in duration of analgesia, because of better sensory spread, 20 mL of ropivacaine 0.5% appears to be the most appropriate dose for peripheral analgesia after TKR. PMID- 15814755 TI - Expectant management of postdural puncture headache increases hospital length of stay and emergency room visits. AB - PURPOSE: This retrospective cohort study examined hospital length of stay (LOS) and emergency room (ER) visits in parturients diagnosed with postdural puncture headache (PDPH) following recognized dural puncture (DP). All cases were managed expectantly. Outcomes were compared with matched controls with uneventful labour epidurals. METHODS: After Ethics Committee approval, the hospital perinatal database was used to identify healthy parturients with recognized DP during labour epidural placement from 1996-2001. Women developing PDPH after expectant management were matched with women with uneventful epidurals and no evidence of PDPH, as well as by parity, delivery mode and admission date. All women delivered term singletons. The primary outcome was LOS (hours) from delivery to discharge. Secondary outcomes included: number (#) nights in hospital, #ER visits for PDPH, epidural blood patch (EBP) timing (pre vs post discharge), EBP location (ward vs ER) and blood volumes used. RESULTS: 26 cases and 26 controls were identified. Precise discharge times were found for 23 cases and 23 controls. In cases, the LOS was increased by a mean of 17 +/- 23.8 (SD) hours; [95% conficence interval (CI) = 8, 26; P = 0.0012] and # nights in hospital was increased by a mean of 0.62 +/- 0.94 nights (95% CI, 0.26, 0.98, P = 0.0027). Nineteen cases (73% 19/26) received at least one EBP. Sixteen cases received at least one EBP prior to discharge with 38% (6/16) returning to ER for re-assessment/repeat EBP. Forty four percent (4/9) of cases without an EBP prior to discharge returned to ER for further assessment/EBP. CONCLUSION: PDPH leads to a significant increase in hospital LOS and ER visits. Studies of preventive therapy are warranted. PMID- 15814756 TI - Hyperbaric treatment of cerebral air embolism in an infant with cyanotic congenital heart disease. AB - PURPOSE: Infants with cyanotic congenital heart disease are at risk for cerebral arterial gas embolism (CAGE) from iv infusion lines. Concern about the hazards and difficulty of caring for such patients inside a hyperbaric chamber may deter referral. We report a complex case in which a small infant was managed successfully using a modified hyperbaric oxygen treatment (HBOT) schedule. CLINICAL FEATURES: A four-month-old 6.19 kg male infant with a recent Glenn shunt for double-outlet right ventricle had a seizure and became unstable immediately after an iv drug infusion. The patient was sedated, intubated and ventilated and dobutamine was commenced. A computerized tomography (CT) scan performed ten hours later demonstrated three intracranial air bubbles. About ten hours later the patient was referred for HBOT which commenced soon afterwards in a multiplace chamber. Since the right-to-left shunt would greatly increase the risk of decompression illness from breathing hyperbaric air HBOT was modified by the use of an abbreviated schedule at reduced pressure. Two 90-min HBOT sessions were administered within 24 hr at 38 feet of sea-water pressure, equivalent to 2.15 atmospheres absolute without any air break. During treatment the infant was ventilated using an Oxford Penlon ventilator. A subsequent CT scan demonstrated the absence of air. After extubation he appeared neurologically intact except for some weakness of the left arm. CONCLUSION: Hyperbaric oxygen may be utilized to treat CAGE in small infants with right-to-left shunt and should be commenced promptly. PMID- 15814757 TI - General anesthesia for patients with Brugada syndrome. A report of six cases. AB - PURPOSE: To review six cases of Brugada syndrome presenting for insertion of a cardioverter-defibrillator under general anesthesia. CLINICAL FEATURES: All patients had a history of syncope, ST segment elevation in the right precordial lead of the electrocardiogram (ECG) which became prominent after a pilsicainide challenge test. Routine monitors, right precordial lead of the ECG and an external defibrillator were installed prior to anesthesia. We administered propofol/midazolam for induction, and propofol/sevoflurane combined with fentanyl for maintenance of anesthesia. Atropine and ephedrine were administered to decrease vagal tone. No ECG change or arrhythmia was observed perioperatively. After the successful implantation of the defibrillator, all patients were discharged without any adverse event. CONCLUSION: By avoiding agents or conditions that may exacerbate Brugada syndrome during anesthesia, we were able to manage the patients uneventfully for implantation of a cardioverter defibrillator. PMID- 15814758 TI - Use of a neck brace minimizes double-lumen tube displacement during patient positioning. AB - PURPOSE: When patients are moved from the supine to the lateral decubitus position, the double-lumen endobronchial tube (DLT) is often displaced. The aim of this study was to determine whether a DLT is displaced when there are no movements of the head and neck. METHODS: One hundred patients scheduled for elective thoracic surgery were randomly divided into control and brace groups. Only a left-sided DLT was used during the study. All patients in the brace group wore a neck collar before the positional change. Using a fibreoptic bronchoscope, the distance from the tracheal opening to the main carina and from the bronchial opening to the bronchial carina was measured in the supine and lateral decubitus positions. RESULTS: Displacement of the DLT (mean +/- SD) during a change from the supine to the lateral decubitus position was greater in the control group (6.3 +/- 5.5 mm in the trachea; 2.4 +/- 3.6 mm in the bronchus) than in the brace group (2.2 +/- 3.9 mm in the trachea; 0.6 +/- 3.1 mm in the bronchus); (P < 0.001). The incidence of clinically significant displacement, greater than 5 mm from the initial correct position, was higher in the control group than in the brace group (48% vs 12%, P < 0.001). CONCLUSION: By restricting head and neck movements with a neck brace, the DLT displacement could be minimized while positioning patients for thoracotomy. The main cause of the DLT displacement during lateral positioning appears to be related to movement of the head and neck. PMID- 15814759 TI - Airway control via the CobraPLA during percutaneous dilatational tracheotomy in five patients. AB - PURPOSE: To evaluate the use of the new supraglottic airway device CobraPLA (CPLA) for performing percutaneous dilatational tracheotomy (PDT) utilizing continuous fibreoptic visualization of the larynx and trachea and uninterrupted airway control. CLINICAL FEATURES: The percutaneous tracheotomies were carried out in five patients (four males and one female; mean age 72 yr, mean height 164.6 cm, mean weight 74 kg) following the Griggs technique under continuous fibreoptic vision and airway control provided by the CPLA. The mean time required for removal of the ETT, positioning of the CPLA, and confirmation of adequate ventilation and cuff seal was 78 sec. The mean time for the entire PDT procedure was six minutes and 57 sec. In one patient a 7-mm tracheostomy cannula was used, and in the other four patients an 8-mm cannula was used. The hemodynamic and respiratory variables remained stable during the entire procedure; there were no adverse events. At no point was there any significant difficulty in placing the CPLA or in providing ventilation or oxygenation. Each procedure could be observed easily in its entirety through the FOB. CONCLUSIONS: This technique can be considered simple and safe because it is video-assisted and ensures a continuous airway control. The CPLA offers several advantages over some other supraglottic devices when performing this surgical procedure. PMID- 15814761 TI - Colorimetric carbon dioxide detector to determine accidental tracheal feeding tube placement. AB - PURPOSE: To determine the accuracy of colorimetric CO2 detection compared to the reference standard two-step radiological confirmation of feeding tube position. METHODS: A prospective study was conducted with patients presenting to a 21-bed medical-surgical intensive care unit. An adapter was developed using an endotracheal tube adapter to connect a colorimetric CO2 detector to a feeding tube in an airtight manner. In part I of the study a feeding tube connected to the colorimeter was inserted into the endotracheal tubes of ten ventilated patients to test the device's ability to detect tracheal placement. In part II patients undergoing feeding tube insertion had tube position confirmed with the colorimeter as well as the reference standard two-step x-ray. RESULTS: In phase I the colorimeter correctly identified tracheal placement in all ten patients. In phase II 93/100 procedures ultimately were eligible; the colorimeter had a sensitivity of 0.88 (95% confidence interval: 0.65-1.00) and specificity of 0.99 (0.97-1.00). The device missed one of the eight tracheal placements. Agreement between the colorimeter and two-step x-ray interpretations was excellent (Kappa 0.86; standard error 0.10). CONCLUSION: We describe a novel, convenient method to confirm esophageal feeding tube placement. The device is easily assembled and inexpensive, but should not be reused. Colorimetric determination of tracheal feeding tube placement with this device has excellent agreement with the reference standard two-step radiological technique. PMID- 15814760 TI - [Propofol sedation allows awake intubation of the difficult airway with the Fastrach LMA]. AB - PURPOSE: To evaluate target controlled infusion anesthesia (TCI) with propofol for conscious intubation [(Ramsay score equal to 3 (RS 3)] through the Fastrach laryngeal mask airway (LMA). METHODS: 17 consenting and unpremedicated patients, who showed criteria for difficult intubation (score developed by Arne et al. > or = 11), were monitored and received supplemental oxygen. Propofol was administered by TCI, with successive targets of 0.6 and 1 microg x mL(-1), while the RS was evaluated: if = 3, LMA intubation was attempted, if < 3 the TCI was increased by steps of 0.2 microg x mL(-1) until an RS of 3 was reached. Local anesthesia (lidocaine 5%) of the oropharynx was carried out at 0.6 and 1 microg x mL(-1), together with local anesthesia of the nasopharynx at 1 microg x mL(-1). A standardized questionnaire evaluated memory of and satisfaction with the technique (score/10) on postoperative day 1. RESULTS: The LMA was inserted in 100% of cases and intubation was successful in 16 out of 17 cases (one failure). The propofol target concentration to obtain a RS of 3 was 1.25 +/- 0.07 microg x mL(-1). Amnesia occurred as soon as the target concentration of propofol exceeded 1 microg x mL(-1). The patients found the technique very satisfactory (median satisfaction score = 9.4/10). Incidents of coughing or nausea were observed in 47% and 5% of cases respectively. There was no oesophageal intubation and no desaturation (SpO2 < 95%). CONCLUSION: Propofol administered by TCI to achieve a RS of 3 allows conscious intubation to be performed through a LMA under satisfactory conditions. A LMA could be a possible alternative to a "conscious" fibroscopy. PMID- 15814763 TI - Grow your own: strategies to develop anesthesia researchers. PMID- 15814762 TI - Preventing pain on injection of rocuronium: two doses of dexmedetomidine. PMID- 15814764 TI - Decrease of the inhibitory effect of lidocaine on trigeminal nerve response by the inflammatory oxidant peroxynitrite. PMID- 15814765 TI - Appropriate waiting time for noncardiac surgery following coronary stent insertion: views of Canadian anesthesiologists [corrected]. PMID- 15814766 TI - S100beta and postcardiac surgery neurological dysfunction: reasons to disregard any link. PMID- 15814767 TI - Intrathoracic positioning of a thoracic epidural catheter inserted via the median approach. PMID- 15814768 TI - Comparison of the accuracy of two different modes of continuous cardiac index measurement following rapid volume infusion. PMID- 15814769 TI - Airway management for postoperative respiratory failure: use of the laryngeal mask airway. PMID- 15814770 TI - Using a CO2 detector to confirm endotracheal intubation in SARS patients. PMID- 15814771 TI - Ventricular fibrillation in the postanesthesia care unit (PACU) secondary to coronary spasm. PMID- 15814772 TI - International Organization for Standardization (ISO) TC85/SC2 WG21: dosimetry for exposures to cosmic radiation in civilian aircraft. PMID- 15814773 TI - Leukocyte populations and steroid receptor expression in human first-trimester decidua; regulation by antiprogestin and prostaglandin E analog. AB - CONTEXT: Progesterone acting via its cognate receptor is critical to maintaining a viable endometrial environment for implantation and pregnancy. During medical termination of pregnancy, the biological effect of progesterone is pharmacologically withdrawn and prostaglandins administered exogenously. Leukocytes within the uterus are the effector cells of an inflammatory response and play important roles in both tissue breakdown and remodeling. OBJECTIVE: The aim of this study was to identify the separate and combined effects of the antiprogestin Mifepristone (single dose, 200 mg) and the prostaglandin E (PGE) analog (gemeprost) on leukocyte populations and steroid receptor expression in human first-trimester decidua. PATIENTS: Eighty women were recruited from the termination of pregnancy service with a gestational age of between 35 and 65 d at the time of surgical termination of pregnancy. MAIN OUTCOME MEASURES: Immunohistochemistry was used to measure macrophage (CD68 +ve), neutrophil (neutrophil elastase +ve), and uterine natural killer cell (CD56 +ve) populations and progesterone (PR(A) and PR(B)), estrogen (ERalpha and ERbeta), and androgen receptor (AR) expression. RESULTS: After administration of both antiprogestin and the PGE analog, macrophage and neutrophil numbers were significantly increased, whereas natural killer cell numbers were unchanged. Antiprogestin and PGE analog coadministration also significantly decreased PR and ERalpha immunoreactivity but had no effect on androgen receptor or ERbeta receptor expression. PGE analog alone was also capable of reducing PR expression. CONCLUSIONS: In this study, we demonstrate that the inflammatory response induced by antiprogestin in combination with PGE analog is accompanied by both increases in macrophages and neutrophils numbers and decreases in PR and ERalpha expression in human first trimester decidua. PMID- 15814774 TI - Association between the current use of low-dose oral contraceptives and cardiovascular arterial disease: a meta-analysis. AB - CONTEXT: The long-term cardiovascular safety of widely used oral contraceptives (OCs) is still debated, and no meta-analysis assesses the modern use of OCs and the associated cardiovascular risks. OBJECTIVE: We aimed to assess the risk of cardiovascular diseases associated with current use of low-dose combined OCs. DATA SOURCES: All studies published between January 1980 and October 2002 were searched using MEDLINE, BIOSIS, and Scientific Citations. STUDY SELECTION: Original studies were selected independently by two investigators (J.P.B., P.A.E.) based on inclusion criteria: low-dose combined OC (<50 mug of ethinyl estradiol); more than 10 cases in low-dose users; clear definition of cases; concurrent controls; and control for age. A third investigator (J.E.N.) adjudicated disagreements. From 2715 identified articles, 14 independent studies were included. DATA EXTRACTION: All data were abstracted by one investigator (J.P.B.) in a systematic manner. Classification of OCs and types of exposure were directly abstracted from studies. Current use was defined as use at the time of the event or within 3 months. Only peer-reviewed studies with definition of events as definite or possible, based on prespecified criteria, were included. DATA SYNTHESIS: The summary risk estimates associated with current use of low dose OCs were 1.84 [95% confidence interval (CI) = 1.38, 2.44] for myocardial infarctions and 2.12 (95% CI = 1.56, 2.86) for ischemic strokes. The overall summary odds ratio for both outcomes was 2.01 (95% CI = 1.63, 2.48). Second generation OCs were associated with a significant increased risk of both myocardial infarction and ischemic stroke events [1.85 (95% CI = 1.03,3.32) and 2.54 (95% CI = 1.96,3.28), respectively]; and third-generation OCs, for ischemic stroke outcome only [2.03 (95% CI = 1.15,3.57)]. CONCLUSIONS: In conclusion, a rigorous meta-analysis of the literature suggests that current use of low-dose OCs significantly increases the risk of both cardiac and vascular arterial events, including a significant risk of vascular arterial complications with third generation OCs. PMID- 15814775 TI - Dirty nails. PMID- 15814776 TI - Osteoporosis and integrins. PMID- 15814777 TI - Tumoral calcinosis: a look into the metabolic mirror of phosphate homeostasis. PMID- 15814778 TI - Persistent neutrophilia as a preceding symptom of pheochromocytoma. PMID- 15814779 TI - A functional role for small-conductance calcium-activated potassium channels in sensory pathways including nociceptive processes. AB - We investigated the role of small-conductance calcium-activated potassium (SK) and intermediate-conductance calcium-activated potassium channels in modulating sensory transmission from peripheral afferents into the rat spinal cord. Subunit specific antibodies reveal high levels of SK3 immunoreactivity in laminas I, II, and III of the spinal cord. Among dorsal root ganglion neurons, both peripherin positive (C-type) and peripherin-negative (A-type) cells show intense SK3 immunoreactivity. Furthermore, dorsal root-stimulated sensory responses recorded in vitro are inhibited when SK channel activity is increased with 1-ethyl-2 benzimidazolinone (1-EBIO). In vivo electrophysiological recordings show that neuronal responses to naturally evoked nociceptive and nonnociceptive stimuli increase after application of the selective SK channel blocker 8,14-diaza 1,7(1,4)-diquinolinacyclotetradecaphanedium di-trifluoroacetate (UCL 1848), indicating that SK channels are normally active in moderating afferent input. Conversely, neuronal responses evoked by mechanical stimuli are inhibited when SK channel activity is increased with 1-EBIO. These effects are reversed by the subsequent application of UCL 1848. Our data demonstrate that SK channels have an important role in controlling sensory input into the spinal cord. PMID- 15814780 TI - PDGF alpha-receptor signal strength controls an RTK rheostat that integrates phosphoinositol 3'-kinase and phospholipase Cgamma pathways during oligodendrocyte maturation. AB - Receptors with tyrosine kinase activity (RTKs) control tissue growth and development in metazoans. How they generate cell-specific responses remains essentially unknown; one model proposes that distinct RTKs activate different second-messenger pathways, whereas a second proposes that all RTKs deliver a generic "go" signal to these pathways that is uniquely interpreted by downstream, cell-specific response competence factors. We examine pathway activation and pathway-specific responses downstream of PDGFalpha receptors, whose expression in the developing CNS identifies oligodendrocyte progenitor cells (OPCs) and whose activation controls OPC proliferation, migration, survival, and maturation. PDGFRalpha-null mice die in utero, and OPCs that emerge before their demise have migration and proliferation defects and rapidly differentiate into postmitotic oligodendrocytes in vitro. OPCs from hemizygous mice also undergo precocious differentiation, indicating a role for PDGFRalpha gene dosage in timing OPC maturation. The rescue of PDGFRalpha-null OPCs with PDGFRalpha transgenes revealed specific roles for the phosphatidylinositol 3-kinase (PI3K) and phospholipase Cgamma (PLCgamma) pathways and a distinct ligand concentration dependence. Activation of the PI3K pathway is required for PDGFRalpha-induced migration, whereas activation of both PI3K and PLCgamma are required for PDGFRalpha-induced proliferation. For proliferation, PI3K activation is required at low ligand concentration, whereas PLCgamma is required at high signal strength. Dose-response studies further demonstrate that PDGFRalpha activates PI3K at low ligand concentrations, whereas PLCgamma is activated at high signal strength. Thus, PDGFRalpha signaling acts like a rheostat rather than generic ON switch, with signal strength dictating pathway activation during OPC maturation. PMID- 15814781 TI - Robustness of burst firing in dissociated purkinje neurons with acute or long term reductions in sodium conductance. AB - Cerebellar Purkinje neurons often generate all-or-none burst firing in response to depolarizing stimuli. Voltage-clamp experiments using action potential waveforms show that burst firing depends on small net inward currents that flow after spikes and reflect the net balance between multiple large currents. Given this, burst firing is surprisingly robust in the face of changes in the magnitude of the underlying currents from cell to cell. We explored the basis of this robustness by examining the effects of reducing the sodium current, the major contributor to the postspike inward current. Burst firing persisted in concentrations of tetrodotoxin that produced half-block of sodium current. This robustness of bursting reflects an acute feedback mechanism whereby waveform changes from the reduced sodium current (reduced spike height and a hyperpolarizing shift in postspike voltage) cause compensatory decreases in postspike potassium currents. In particular, reduced spike height reduces calcium entry and subsequent calcium-activated potassium current, and the hyperpolarizing shift in postspike voltage speeds deactivation of Kv3-like potassium channels. Other experiments examined bursting in Na(v)1.6-/- mice, in which sodium current density is reduced in the long term. Under these circumstances, there was upregulation of both T-type and P-type calcium current and a change in the balance of calcium current and calcium-activated potassium current such that their net influence shifted from being inhibitory during bursts in wild-type neurons to excitatory during bursts from Na(v)1.6-/- mutant neurons. Thus, Purkinje neurons have both acute and long-term feedback mechanisms that serve to maintain burst firing when voltage-dependent sodium conductance is reduced. PMID- 15814782 TI - Dendrodendritic synaptic signals in olfactory bulb granule cells: local spine boost and global low-threshold spike. AB - In the mammalian olfactory bulb, axonless granule cells process synaptic input and output reciprocally within large spines. The nature of the calcium signals that underlie the presynaptic and postsynaptic function of these spines is mostly unknown. Using two-photon imaging in acute rat brain slices and glomerular stimulation of mitral/tufted cells, we observed two forms of action potential independent synaptic Ca2+ signals in granule cell dendrites. Weak activation of mitral/tufted cells produced stochastic Ca2+ transients in individual granule cell spines. These transients were strictly localized to the spine head, indicating a local passive boosting or spine spike. Ca2+ sources for these local synaptic events included NMDA receptors, voltage-dependent calcium channels, and Ca2+-induced Ca2+ release from internal stores. Stronger activation of mitral/tufted cells produced a low-threshold Ca2+ spike (LTS) throughout the granule cell apical dendrite. This global spike was mediated by T-type Ca2+ channels and represents a candidate mechanism for subthreshold lateral inhibition in the olfactory bulb. The coincidence of local input and LTS in the spine resulted in summation of local and global Ca2+ signals, a dendritic computation that could endow granule cells with subthreshold associative plasticity. PMID- 15814783 TI - Visual control of burst priming in the anesthetized lateral geniculate nucleus. AB - Thalamic relay cells fire bursts of action potentials. Once a long hyperpolarization "primes" (deinactivates) the T-type calcium channel, a depolarizing input will "trigger" a calcium spike with a burst of action potentials. During sleep, bursts are frequent, rhythmic, and nonvisual. Bursts have been observed in alert animals, and burst timing is known to carry visual information under light anesthesia. We extend this finding by showing that bursts without visual triggers are rare. Nevertheless, if the channel were primed at random with respect to the stimulus, then bursts would have the same visual significance as single spikes. We find, however, that visual signals influence when the channel is primed. First, natural time-varying stimuli evoke more bursts than white noise. Second, specific visual stimuli reproducibly elicit bursts, whereas others reliably elicit single spikes. Therefore, visual information is encoded by the selective tagging of some responses as bursts. The visual information attributable to visual priming (as distinct from the information attributable to visual triggering of the bursts) was two bits per burst on average. Although bursts are reportedly rare in alert animals, this must be investigated as a function of visual stimulus. Moreover, we propose methods to measure the extent of both visual triggering and visual priming of bursts. Whether or not bursts are rare, our methods could help determine whether bursts in alert animals carry a distinct visual signal. PMID- 15814784 TI - Transgenic mouse model of tau pathology in astrocytes leading to nervous system degeneration. AB - Filamentous tau inclusions in neurons and glia are neuropathological hallmarks of sporadic and familial tauopathies. Because tau gene mutations are pathogenic for the autosomal dominant tauopathy "frontotemporal dementia and parkinsonism linked to chromosome 17," tau abnormalities are implicated directly in the onset and/or progression of disease. Although filamentous tau aggregates are acknowledged to play roles in degenerative mechanisms resulting in neuron loss, the contributions of glial tau pathology to neurodegeneration remain essentially unexplored. To begin to elucidate the role of glial pathology in tauopathies, we generated a transgenic (Tg) mouse model of astrocytic tau pathology by expressing the human tau protein driven by the glial fibrillary acidic protein (GFAP) promoter. Whereas endogenous tau was not detected in astrocytes of control mice, in GFAP/tau Tg mice there was robust astrocytic tau expression that was associated with a redistribution of the GFAP network. Subsequently, there was an age dependent accumulation of tau pathology in astrocytes that was Gallyas and variably thioflavine S positive as observed in many tauopathies. The tau pathology in these Tg mice was abnormally phosphorylated, ubiquitinated, and filamentous, and the emergence of this pathology coincided with accumulation of insoluble tau protein. Furthermore, in regions with robust astrocytic tau pathology, there was mild blood- brain barrier disruption, induction of low molecular-weight heat shock proteins, and focal neuron degeneration. Thus, these Tg mice recapitulate key features of astrocytic pathology observed in human tauopathies and demonstrate functional consequences of this pathology including neuron degeneration in the absence of neuronal tau inclusions. PMID- 15814785 TI - Spinal G-protein-gated potassium channels contribute in a dose-dependent manner to the analgesic effect of mu- and delta- but not kappa-opioids. AB - Opioids can evoke analgesia by inhibiting neuronal targets in either the brain or spinal cord, and multiple presynaptic and postsynaptic inhibitory mechanisms have been implicated. The relative significance of presynaptic and postsynaptic inhibition to opioid analgesia is essentially unknown, as are the identities and relevant locations of effectors mediating opioid actions. Here, we examined the distribution of G-protein-gated potassium (GIRK) channels in the mouse spinal cord and measured their contribution to the analgesia evoked by spinal administration of opioid receptor-selective agonists. We found that the GIRK channel subunits GIRK1 and GIRK2 were concentrated in the outer layer of the substantia gelatinosa of the dorsal horn. GIRK1 and GIRK2 were found almost exclusively in postsynaptic membranes of putative excitatory synapses, and a significant degree of overlap with the mu-opioid receptor was observed. Although most GIRK subunit labeling was perisynaptic or extrasynaptic, GIRK2 was found occasionally within the synaptic specialization. Genetic ablation or pharmacologic inhibition of spinal GIRK channels selectively blunted the analgesic effect of high but not lower doses of the mu-opioid receptor-selective agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin. Dose-dependent contributions of GIRK channels to the analgesic effects of the -opioid receptor-selective agonists Tyr-D-Ala-Phe-Glu-Val-Val-Gly amide and [D-Pen(2,5)]-enkephalin were also observed. In contrast, the analgesic effect of the agonist (trans)-3,4 dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl] benzeneacetamide methanesulfonate hydrate was preserved despite the absence of GIRK channels. We conclude that the activation of postsynaptic GIRK1 and/or GIRK2-containing channels in the spinal cord dorsal horn represents a powerful, albeit relatively insensitive, means by which intrathecal mu- and -selective opioid agonists evoke analgesia. PMID- 15814786 TI - Shank expression is sufficient to induce functional dendritic spine synapses in aspiny neurons. AB - Shank proteins assemble glutamate receptors with their intracellular signaling apparatus and cytoskeleton at the postsynaptic density. Whether Shank plays a role in spinogenesis and synaptogenesis remained unclear. Here, we report that knock-down of Shank3/prolinerich synapse-associated protein-2 by RNA interference reduces spine density in hippocampal neurons. Moreover, transgene expression of Shank 3 is sufficient to induce functional dendritic spines in aspiny cerebellar neurons. Transfected Shank protein recruits functional glutamate receptors, increases the number and size of synaptic contacts, and increases amplitude, frequency, and the AMPA component of miniature EPSCs, similar to what is observed during synapse developmental maturation. Mutation/deletion approaches indicate that these effects require interactions of Shank3 with the glutamate receptor complex. Consistent with this observation, chronic treatment with glutamate receptor antagonists alters maturation of the Shank3-induced spines. These results strongly suggest that Shank proteins and the associated glutamate receptors participate in a concerted manner to form spines and functional synapses. PMID- 15814787 TI - Selective modulation of neuronal nicotinic acetylcholine receptor channel subunits by Go-protein subunits. AB - G-protein modulation of neuronal nicotinic acetylcholine receptor (nAChR) channels in rat intrinsic cardiac ganglia was examined using dialyzed whole-cell and excised membrane patch-recording configurations. Cell dialysis with GTPgammaS increased the agonist affinity of nAChRs, resulting in a potentiation of nicotine evoked whole-cell currents at low concentrations. ACh- and nicotine-evoked current amplitudes were increased approximately twofold in the presence of GTPgammaS. In inside-out membrane patches, the open probability (NP(o)) of nAChR mediated unitary currents was reversibly increased fourfold after bath application of 0.2 mm GTPgammaS relative to control but was unchanged in the presence of GDPbetaS. The modulation of nAChR-mediated whole-cell currents was agonist specific; currents evoked by the cholinergic agonists ACh, nicotine, and 1,1-dimethyl-4-phenylpiperazinium iodide, but not cytisine or choline, were potentiated in the presence of GTPgammaS. The direct interaction between G protein subunits and nAChRs was examined by bath application of either G(o)alpha or Gbetagamma subunits to inside-out membrane patches and in glutathione S transferase pull-down and coimmunoprecipitation experiments. Bath application of 50 nm Gbetagamma increased the open probability of ACh-activated single-channel currents fivefold, whereas G(o)alpha (50 nm) produced no significant increase in NP(o). Neuronal nAChR subunits alpha3-alpha5 and beta2 exhibited a positive interaction with G(o)alpha and Gbetagamma, whereas beta4 and alpha7 failed to interact with either of the G-protein subunits. These results provide evidence for a direct interaction between nAChR and G-protein subunits, underlying the increased open probability of ACh-activated single-channel currents and potentiation of nAChR-mediated whole-cell currents in parasympathetic neurons of rat intrinsic cardiac ganglia. PMID- 15814788 TI - Proopiomelanocortin neurons in nucleus tractus solitarius are activated by visceral afferents: regulation by cholecystokinin and opioids. AB - The nucleus tractus solitarius (NTS) receives dense terminations from cranial visceral afferents, including those from the gastrointestinal (GI) system. Although the NTS integrates peripheral satiety signals and relays this signal to central feeding centers, little is known about which NTS neurons are involved or what mechanisms are responsible. Proopiomelanocortin (POMC) neurons are good candidates for GI integration, because disruption of the POMC gene leads to severe obesity and hyperphagia. Here, we used POMC-enhanced green fluorescent protein (EGFP) transgenic mice to identify NTS POMC neurons. Intraperitoneal administration of cholecystokinin (CCK) induced c-fos gene expression in NTS POMC EGFP neurons, suggesting that they are activated by afferents stimulated by the satiety hormone. We tested the synaptic relationship of these neurons to visceral afferents and their modulation by CCK and opioids using patch recordings in horizontal brain slices. Electrical activation of the solitary tract (ST) evoked EPSCs in NTS POMC-EGFP neurons. The invariant latencies, low failure rates, and substantial paired-pulse depression of the ST-evoked EPSCs indicate that NTS POMC EGFP neurons are second-order neurons directly contacted by afferent terminals. The EPSCs were blocked by the glutamate antagonist 2,3-dihydroxy-6-nitro-7 sulfonyl-benzo[f]quinoxaline. CCK increased the amplitude of the ST-stimulated EPSCs and the frequency of miniature EPSCs, effects attenuated by the CCK1 receptor antagonist lorglumide. In contrast, the orexigenic opioid agonists [D Ala(2), N-Me-Phe(4), Gly-ol(5)]-enkephalin and met-enkephalin inhibited both ST stimulated EPSCs and the frequency of miniature EPSCs. These findings identify a potential satiety pathway in which visceral afferents directly activate NTS POMC EGFP neurons with excitatory inputs that are appropriately modulated by appetite regulators. PMID- 15814789 TI - Continuous and overlapping expression domains of odorant receptor genes in the olfactory epithelium determine the dorsal/ventral positioning of glomeruli in the olfactory bulb. AB - In mammals, olfactory signals received by odorant receptors (ORs) in the olfactory epithelium (OE) are converted to a topographical map of activated glomeruli in the olfactory bulb (OB). It has been reported that the OE can be divided into four topographically distinct zones and that olfactory sensory neurons (OSNs) expressing a particular OR gene are randomly distributed within one zone. Here, we analyzed 80 different class II OR genes for their expression patterns in the OE by in situ hybridization. It was found that the expression area in the OE does not always fit into one of the four conventional zones. Expression areas are specific to each OR gene and are arranged in an overlapping and continuous manner in the OE. We also analyzed a spatial relationship between the OE and the OB for OSN projection. Our transgenic as well as DiI retrograde staining experiments demonstrated that the dorsal/ventral arrangement of glomeruli in the OB is correlated with the expression areas of corresponding ORs along the dorsomedial/ventrolateral axis in the OE. The present study indicates that the OR gene choice may be more restricted by the OSN location in the OE than what has been thought. PMID- 15814790 TI - Attentional modulation of learning-related repetition attenuation effects in human parahippocampal cortex. AB - Two of the most fundamental processes in biological vision are attention and learning. Attention actively selects and enhances visual information that is most relevant to behavior. Learning enables the visual system to benefit from perceptual experience. The amount of visual information to learn is infinite; however, top-down control mechanisms must somehow regulate learning to achieve an adaptive balance between plasticity and stability in neural circuitry. Functional magnetic resonance imaging (fMRI) can measure learning-related changes in neural activity to previously viewed perceptual stimuli. Described variably as the repetition suppression or adaptation effect, the attenuation in neural activity to repeated stimuli versus novel stimuli provides a marker for stimuli-specific perceptual processing and memory. One important issue concerns whether repetition attenuation is automatic or not, and recent work has begun to show that it is sensitive to task demands. Accordingly, the present study further examined how attention controls the attenuated response to repeated stimuli, specifically testing whether attention is important for initial encoding, for the expression of memory traces, or for both encoding and expression. To manipulate attention, we used overlapping scene and face images and asked subjects to attend to either category. fMRI revealed significant attenuation in the parahippocampal place area for only the repeated scenes that were attended both during the initial presentation and during repetition. Thus, attention actively governs when neuronal activity is attenuated to repeated perceptual input, and such attention is important during both initial encoding and subsequent expression of the learned information. PMID- 15814791 TI - Modeling spontaneous activity in the developing spinal cord using activity dependent variations of intracellular chloride. AB - We investigated how spontaneous activity is generated in developing, hyperexcitable networks. We focused our study on the embryonic chick spinal cord, a preparation that exhibits rhythmic discharge on multiple timescales: slow episodes (lasting minutes) and faster intraepisode cycling (approximately 1 Hz frequency). For this purpose, we developed a mean field model of a recurrent network with slow chloride dynamics and a fast depression variable. We showed that the model, in addition to providing a biophysical mechanism for the slow dynamics, was able to account for the experimentally observed activity. The model made predictions on how interval and duration of episodes are affected when changing chloride-mediated synaptic transmission or chloride flux across cell membrane. These predictions guided experiments, and the model results were compared with experimental data obtained with electrophysiological recordings. We found agreement when transmission was affected through changes in synaptic conductance and good qualitative agreement when chloride flux was varied through changes in external chloride concentration or in the rate of the Na+-K+-2Cl- cotransporter. Furthermore, the model made predictions about the time course of intracellular chloride concentration and chloride reversal potential and how these are affected by changes in synaptic conductance. Based on the comparison between modeling and experimental results, we propose that chloride dynamics could be an important mechanism in rhythm generation in the developing chick spinal cord. PMID- 15814792 TI - Secreted semaphorins modulate synaptic transmission in the adult hippocampus. AB - Modulation of synaptic activity is critical for neural circuit function and behavior. The semaphorins are a large, phylogenetically conserved protein family with important roles in neural development. However, semaphorin function in the adult brain has yet to be determined. Here, we show that the coreceptors for secreted semaphorins, the neuropilins, are found at synapses and localize to molecular layers of the adult mouse hippocampus and accessory olfactory cortex. Moreover, application of the secreted semaphorin Sema3F to acute hippocampal slices modulates both the frequency and amplitude of miniature EPSCs in granule cells of the dentate gyrus and pyramidal neurons of CA1. Finally, we show that mice lacking Sema3F are prone to seizures. These results suggest a novel role for semaphorins as synaptic modulators and illustrate the diverse repertoire of these guidance cues in both the formation and function of neural circuits. PMID- 15814793 TI - Apolipoprotein E genotype regulates amyloid-beta cytotoxicity. AB - The epsilon4 allele of apolipoprotein E (ApoE) is a risk factor for Alzheimer's disease (AD), whereas the epsilon2 allele may be relatively protective. Both alleles are risk factors for cerebral amyloid angiopathy (CAA)-related hemorrhages. CAA is associated with degeneration of smooth muscle cells and pericytes. Previously, we described that synthetic amyloid-beta1-40 peptide (Abeta1-40) with the 22Glu--> Gln "Dutch" mutation caused pericyte death in vitro by a mechanism that involves Abeta fibril-like assembly at the cell surface. It is known that ApoE binds to Abeta and may modify its biological activities. In the present study, we evaluated the effect of ApoE on Abeta-mediated toxicity of cerebrovascular cells. We observed that cultured cells with an epsilon4/epsilon4 genotype were more vulnerable to Abeta than cultures with an epsilon3/epsilon3 or epsilon3/epsilon4 genotype. The one cell culture with the epsilon2/epsilon3 genotype was relatively resistant to Abeta compared with other cultures. Furthermore, we observed a dose-dependent protective effect of native ApoE against Abeta-mediated toxicity of cerebrovascular cells and, in addition, ApoE epsilon2/epsilon3 cells secreted more ApoE protein compared with cells with other ApoE genotypes, in particular, compared with epsilon4/epsilon4 cells. Thus, the disparity between ApoE genotype and Abeta-mediated toxicity might be related to differences in the cellular capacity to secrete ApoE. The present data suggest that one mechanism by which ApoE may alter the risk for AD is a genotype dependent regulation of Abeta cytotoxicity, possibly via variations in its secretion levels, whereby extracellular ApoE may bind to Abeta and thereby modify Abeta-mediated cell death. PMID- 15814794 TI - Plexin-a4 mediates axon-repulsive activities of both secreted and transmembrane semaphorins and plays roles in nerve fiber guidance. AB - It has been proposed that four members of the plexin A subfamily (plexin-As; plexin-A1, -A2, -A3, and -A4) and two neuropilins (neuropilin-1 and neuropilin-2) form complexes and serve as receptors for class 3 secreted semaphorins (Semas), potent neural chemorepellents. The roles of given plexin-As in semaphorin signaling and axon guidance, however, are mostly unknown. Here, to elucidate functions of plexin-A4 in semaphorin signaling and axon guidance events in vivo, we generated plexin-A4 null mutant mice by targeted disruption of the plexin-A4 gene. Plexin-A4 mutant mice were defective in the trajectory and projection of peripheral sensory axons and sympathetic ganglion (SG) axons and the formation of the anterior commissure and the barrels. The defects in peripheral sensory and SG axons were fundamentally related to those of neuropilin-1 or Sema3A mutant embryos reported but were more moderate than the phenotype in these mutants. The growth cone collapse assay showed that dorsal root ganglion axons and SG axons of plexin-A4 mutant embryos partially lost their responsiveness to Sema3A. These results suggest that plexin-A4 plays roles in the propagation of Sema3A activities and regulation of axon guidance and that other members of the plexin-A subfamily are also involved in the propagation of Sema3A activities. Plexin-A4 deficient SG axons did not lose their responsiveness to Sema3F, suggesting that plexin-A4 serves as a Sema3A-specific receptor, at least in SG axons. In addition, the present study showed that plexin-A4 bound class 6 transmembrane semaphorins, Sema6A and Sema6B, and mediated their axon-repulsive activities, independently of neuropilin-1. Our results imply that plexin-A4 mediates multiple semaphorin signals and regulates axon guidance in vivo. PMID- 15814795 TI - Astrocytes regulate inhibitory synapse formation via Trk-mediated modulation of postsynaptic GABAA receptors. AB - Astrocytes promote the formation and function of excitatory synapses in the CNS. However, whether and how astrocytes modulate inhibitory synaptogenesis are essentially unknown. We asked whether astrocytes regulate the formation of inhibitory synapses between hippocampal neurons during maturation in vitro. Neuronal coculture with astrocytes or treatment with astrocyte-conditioned medium (ACM) increased the number of inhibitory presynaptic terminals, the frequency of miniature IPSCs, and the number and synaptic localization of GABA(A) receptor (GABA(A)R) clusters during the first 10 d in vitro. We asked whether neurotrophins, which are potent modulators of inhibitory synaptic structure and function, mediate the effects of astrocytes on inhibitory synapses. ACM from BDNF or tyrosine receptor kinase B (TrkB)-deficient astrocytes increased inhibitory presynaptic terminals and postsynaptic GABA(A)R clusters in wild-type neurons, suggesting that BDNF and TrkB expression in astrocytes is not required for these effects. In contrast, although the increase in the number of inhibitory presynaptic terminals persisted, no increase was observed in postsynaptic GABA(A)R clusters after ACM treatment of hippocampal neurons lacking BDNF or TrkB. These results suggest that neurons, not astrocytes, are the relevant source of BDNF and are the site of TrkB activation required for postsynaptic GABA(A)R modulation. These data also suggest that astrocytes may modulate postsynaptic development indirectly by stimulating Trk signaling between neurons. Together, these data show that astrocytes modulate inhibitory synapse formation via distinct presynaptic and postsynaptic mechanisms. PMID- 15814796 TI - Inhibition by spinal mu- and delta-opioid agonists of afferent-evoked substance P release. AB - Opioid mu- and delta-receptors are present on the central terminals of primary afferents, where they are thought to inhibit neurotransmitter release. This mechanism may mediate analgesia produced by spinal opiates; however, when they used neurokinin 1 receptor (NK1R) internalization as an indicator of substance P release, Trafton et al. (1999) noted that this evoked internalization was altered only modestly by morphine delivered intrathecally at spinal cord segment S1-S2. We reexamined this issue by studying the effect of opiates on NK1R internalization in spinal cord slices and in vivo. In slices, NK1R internalization evoked by dorsal root stimulation at C-fiber intensity was abolished by the mu agonist [D-Ala2, N-Me-Phe4, Gly-ol5]-enkephalin (DAMGO) (1 microM) and decreased by the delta agonist [D-Phe2,5]-enkephalin (DPDPE) (1 microM). In vivo, hindpaw compression induced NK1R internalization in ipsilateral laminas I-II. This evoked internalization was significantly reduced by morphine (60 nmol), DAMGO (1 nmol), and DPDPE (100 nmol), but not by the kappa agonist trans-(1S,2S)-3,4-dichloro-N-mathyl-N-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetamide hydrochloride (200 nmol), delivered at spinal cord segment L2 using intrathecal catheters. These doses of the mu and delta agonists were equi analgesic as measured by a thermal escape test. Lower doses neither produced analgesia nor inhibited NK1R internalization. In contrast, morphine delivered by percutaneous injections at S1-S2 had only a modest effect on thermal escape, even at higher doses. Morphine decreased NK1R internalization after systemic delivery, but at a dose greater than that necessary to produce equivalent analgesia. All effects were reversed by naloxone. These results indicate that lumbar opiates inhibit noxious stimuli-induced neurotransmitter release from primary afferents at doses that are confirmed behaviorally as analgesic. PMID- 15814797 TI - Stimulus dependence of neuronal correlation in primary visual cortex of the macaque. AB - Nearby cortical neurons often have correlated trial-to-trial response variability, and a significant fraction of their spikes occur synchronously. These two forms of correlation are both believed to arise from common synaptic input, but the origin of this input is unclear. We investigated the source of correlated responsivity by recording from pairs of single neurons in primary visual cortex of anesthetized macaque monkeys and comparing correlated variability and synchrony for spontaneous activity and activity evoked by stimuli of different orientations and contrasts. These two stimulus manipulations would be expected to have different effects on the cortical pool providing input to the recorded pair: changing stimulus orientation should recruit different populations of cells, whereas changing stimulus contrast affects primarily the relative strength of sensory drive and ongoing cortical activity. Consistent with this predicted difference, we found that correlation was affected by these stimulus manipulations in different ways. Synchrony was significantly stronger for orientations that drove both neurons well than for those that did not, but correlation on longer time scales was orientation independent. Reducing stimulus contrast resulted in a decrease in the temporal precision of synchronous firing and an enhancement of correlated response variability on longer time scales. Our results thus suggest that correlated responsivity arises from mechanisms operating at two distinct timescales: one that is orientation tuned and that determines the strength of temporally precise synchrony, and a second that is contrast sensitive, of low temporal frequency, and present in ongoing cortical activity. PMID- 15814798 TI - Repeated cocaine administration increases voltage-sensitive calcium currents in response to membrane depolarization in medial prefrontal cortex pyramidal neurons. AB - The medial prefrontal cortex (mPFC) plays a critical role in cocaine addiction. However, evidence to elucidate how the mPFC is functionally involved in cocaine addiction remains incomplete. Recent studies have revealed that repeated cocaine administration induces various neuroadaptations in pyramidal mPFC neurons, including a reduction in voltage-gated K+ currents (VGKCs) and a possible increase in voltage-sensitive Ca2+ currents (I(Ca)). Here, we performed both current-clamp recordings in brain slices and voltage-clamp recordings in freshly dissociated cells to determine whether I(Ca) is altered in mPFC pyramidal neurons after chronic cocaine treatment with a short-term or long-term withdrawal. In addition, a critical role of VGKCs in regulating the generation of Ca2+ plateau potential was also studied in mPFC neurons. Repeated cocaine administration significantly prolonged the duration of evoked Ca2+ plateau potentials and increased the whole-cell I(Ca) in mPFC neurons after a 3 d withdrawal. Selective blockade of L-type Ca2+ channels by nifedipine not only significantly increased the threshold but also reduced the duration and amplitude of Ca2+ plateau potentials in both saline- and cocaine-withdrawn mPFC neurons. However, there was no significant difference in the increased threshold, reduced duration, and decreased amplitude of Ca2+ potentials between saline- and cocaine-withdrawn neurons after blockade of L-type Ca2+ channels. Moreover, an increase in amplitude was also observed, whereas the prolonged duration persisted, in Ca2+ potentials after 2-3 weeks of withdrawal. These findings indicate that chronic exposure to cocaine facilitates the responsiveness of I(Ca), particularly via the activated L-type Ca2+ channels, to excitatory stimuli in rat mPFC pyramidal neurons. PMID- 15814799 TI - Spectral edge sensitivity in neural circuits of the dorsal cochlear nucleus. AB - One possible function of the dorsal cochlear nucleus (DCN) is discrimination of head-related transfer functions (HRTFs), spectral cues used for vertical sound localization. Recent psychophysical and physiological studies suggest that steep, rising spectral edges may be the features used to identify HRTFs. Here we showed, using notch noise and noise band stimuli presented over a range of frequencies, that a subclass of DCN type IV neurons responded with a response peak when the rising spectral edge of a notch or band was aligned near best frequency (BF). This edge sensitivity was correlated with weak or inhibited responses to broadband noise and inhibition in receptive fields at frequencies below BF. Some aspects of the inhibition shaping the response peak, namely inhibition to rising edges below BF and to falling edges at BF, could be explained by the properties of type II interneurons with BFs below those of the type IV neurons. However, many type IV neurons also showed inhibitory responses with the rising spectral edge just above BF, and these responses could not be reproduced by current models of DCN circuitry. Therefore, a new component of the DCN circuit is needed to fully explain the responses to rising spectral edges. This shaping of edge sensitivity by inhibition to rising spectral edges both below and above BF suggests the specialization of DCN for spectral edge coding along the tonotopic gradient. PMID- 15814800 TI - Impeded interaction between Schwann cells and axons in the absence of laminin alpha4. AB - The Schwann cell basal lamina (BL) is required for normal myelination. Loss or mutations of BL constituents, such as laminin-2 (alpha2beta1gamma1), lead to severe neuropathic diseases affecting peripheral nerves. The function of the second known laminin present in Schwann cell BL, laminin-8 (alpha4beta1gamma1), is so far unknown. Here we show that absence of the laminin alpha4 chain, which distinguishes laminin-8 from laminin-2, leads to a disturbance in radial sorting, impaired myelination, and signs of ataxia and proprioceptive disturbances, whereas the axonal regenerative capacity is not influenced. In vitro studies show poor axon growth of spinal motoneurons on laminin-8, whereas it is extensive on laminin-2. Schwann cells, however, extend longer processes on laminin-8 than on laminin-2, and, in contrast to the interaction with laminin-2, solely use the integrin receptor alpha6beta1 in their interaction with laminin-8. Thus, laminin 2 and laminin-8 have different critical functions in peripheral nerves, mediated by different integrin receptors. PMID- 15814801 TI - Matrix metalloproteinase-3: a novel signaling proteinase from apoptotic neuronal cells that activates microglia. AB - Microglial activation and inflammation are associated with progressive neuronal apoptosis in neurodegenerative human brain disorders. We sought to investigate molecular signaling mechanisms that govern activation of microglia in apoptotic neuronal degeneration. We report here that the active form of matrix metalloproteinase-3 (MMP-3) was released into the serum-deprived media (SDM) of PC12 cells and other media of apoptotic neuronal cells within 2-6 h of treatment of the cells, and SDM and catalytic domain of recombinant MMP-3 (cMMP-3) activated microglia in primary microglia cultures as well as BV2 cells, a mouse microglia cell line. Both SDM and cMMP-3 induced generation of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), IL-1beta, and interleukin-1 receptor antagonist but not IL-12 and inducible nitric oxide synthase, which are readily induced by lipopolysaccharide, in microglia, suggesting that there is a characteristic pattern of microglial cytokine induction by apoptotic neurons. Neither glial cell line-derived neurotrophic factor nor anti-inflammatory cytokines, such as IL-10 and transforming growth factor-beta1, were induced. SDM and cMMP-3 extensively released TNF-alpha from microglia and activated the nuclear factor-kappaB pathway, and these microglial responses were totally abolished by preincubation with an MMP-3 inhibitor, NNGH [N-isobutyl-N-(4 methoxyphenylsulfonyl)-glycylhydroxamic acid]. MMP-3-mediated microglial activation mostly depended on ERK (extracellular signal-regulated kinase) phosphorylation but not much on either JNK (c-Jun N-terminal protein kinase) or p38 activation. Conditioned medium of SDM- or cMMP-3-activated BV2 cells caused apoptosis of PC12 cells. These results strongly suggest that the distinctive signal of neuronal apoptosis is the release of active form of MMP-3 that activates microglia and subsequently exacerbates neuronal degeneration. Therefore, the release of MMP-3 from apoptotic neurons may play a major role in degenerative human brain disorders, such as Parkinson's disease. PMID- 15814802 TI - Rapid upregulation of alpha7 nicotinic acetylcholine receptors by tyrosine dephosphorylation. AB - Alpha7 nicotinic acetylcholine receptors (nAChRs) modulate network activity in the CNS. Thus, functional regulation of alpha7 nAChRs could influence the flow of information through various brain nuclei. It is hypothesized here that these receptors are amenable to modulation by tyrosine phosphorylation. In both Xenopus oocytes and rat hippocampal interneurons, brief exposure to a broad-spectrum protein tyrosine kinase inhibitor, genistein, specifically and reversibly potentiated alpha7 nAChR-mediated responses, whereas a protein tyrosine phosphatase inhibitor, pervanadate, caused depression. Potentiation was associated with an increased expression of surface alpha7 subunits and was not accompanied by detectable changes in receptor open probability, implying that the increased function results from an increased number of alpha7 nAChRs. Soluble N ethylmaleimide-sensitive factor attachment protein receptor-mediated exocytosis was shown to be a plausible mechanism for the rapid delivery of additional alpha7 nAChRs to the plasma membrane. Direct phosphorylation/dephosphorylation of alpha7 subunits was unlikely because mutation of all three cytoplasmic tyrosine residues did not prevent the genistein-mediated facilitation. Overall, these data are consistent with the hypothesis that the number of functional cell surface alpha7 nAChRs is controlled indirectly via processes involving tyrosine phosphorylation. PMID- 15814804 TI - Is recombinant FVIIa the magic bullet in the treatment of major bleeding? PMID- 15814805 TI - Contamination of anaesthetic gases with nitric oxide and its influence on oxygenation. PMID- 15814803 TI - Presynaptic Na+ channels: locus, development, and recovery from inactivation at a high-fidelity synapse. AB - Na+ channel recovery from inactivation limits the maximal rate of neuronal firing. However, the properties of presynaptic Na+ channels are not well established because of the small size of most CNS boutons. Here we study the Na+ currents of the rat calyx of Held terminal and compare them with those of postsynaptic cells. We find that presynaptic Na+ currents recover from inactivation with a fast, single-exponential time constant (24 degrees C, tau of 1.4-1.8 ms; 35 degrees C, tau of 0.5 ms), and their inactivation rate accelerates twofold during development, which may contribute to the shortening of the action potential as the terminal matures. In contrast, recordings from postsynaptic cells in brainstem slices, and acutely dissociated, reveal that their Na+ currents recover from inactivation with a double-exponential time course (tau(fast) of 1.2-1.6 ms; tau(slow) of 80-125 ms; 24 degrees C). Surprisingly, confocal immunofluorescence revealed that Na+ channels are mostly absent from the calyx terminal but are instead highly concentrated in an unusually long (approximately 20-40 microm) unmyelinated axonal heminode. Outside-out patch recordings confirmed this segregation. Expression of Na(v)1.6 alpha-subunit increased during development, whereas the Na(v)1.2alpha-subunit was not present. Serial EM reconstructions also revealed a long pre-calyx heminode, and biophysical modeling showed that exclusion of Na+ channels from the calyx terminal produces an action potential waveform with a shorter half-width. We propose that the high density and polarized locus of Na+ channels on a long heminode are critical design features that allow the mature calyx of Held terminal to fire reliably at frequencies near 1 kHz. PMID- 15814806 TI - Anaesthetic management of high-risk cardiac patients undergoing thoracic surgery with the support of intra-aortic balloon pump. PMID- 15814807 TI - Who is at increased risk of pulmonary aspiration? PMID- 15814808 TI - Failed intubation in obstetrics: has the incidence changed recently? PMID- 15814809 TI - Explosions in anaesthesia. PMID- 15814810 TI - The superficial ulnar artery--a potential hazard in patients with difficult venous access. PMID- 15814811 TI - Tandem oligonucleotide synthesis using linker phosphoramidites. AB - Multiple oligonucleotides of the same or different sequence, linked end-to-end in tandem can be synthesized in a single automated synthesis. A linker phosphoramidite [R. T. Pon and S. Yu (2004) Nucleic Acids Res., 32, 623-631] is added to the 5'-terminal OH end of a support-bound oligonucleotide to introduce a cleavable linkage (succinic acid plus sulfonyldiethanol) and the 3'-terminal base of the new sequence. Conventional phosphoramidites are then used for the rest of the sequence. After synthesis, treatment with ammonium hydroxide releases the oligonucleotides from the support and cleaves the linkages between each sequence. Mixtures of one oligonucleotide with both 5'- and 3'-terminal OH ends and other oligonucleotides with 5'-phosphorylated and 3'-OH ends are produced, which are deprotected and worked up as a single product. Tandem synthesis can be used to make pairs of PCR primers, sets of cooperative oligonucleotides or multiple copies of the same sequence. When tandem synthesis is used to make two self complementary sequences, double-stranded structures spontaneously form after deprotection. Tandem synthesis of oligonucleotide chains containing up to six consecutive 20mer (120 bases total), various trinucleotide codons and primer pairs for PCR, or self-complementary strands for in situ formation of double stranded DNA fragments has been demonstrated. PMID- 15814812 TI - Experimental RNomics in Aquifex aeolicus: identification of small non-coding RNAs and the putative 6S RNA homolog. AB - By an experimental RNomics approach, we have generated a cDNA library from small RNAs expressed from the genome of the hyperthermophilic bacterium Aquifex aeolicus. The library included RNAs that were antisense to mRNAs and tRNAs as well as RNAs encoded in intergenic regions. Substantial steady-state levels in A.aeolicus cells were confirmed for several of the cloned RNAs by northern blot analysis. The most abundant intergenic RNA of the library was identified as the 6S RNA homolog of A.aeolicus. Although shorter in size (150 nt) than its gamma proteobacterial homologs (approximately 185 nt), it is predicted to have the most stable structure among known 6S RNAs. As in the gamma-proteobacteria, the A.aeolicus 6S RNA gene (ssrS) is located immediately upstream of the ygfA gene encoding a widely conserved 5-formyltetrahydrofolate cyclo-ligase. We identifed novel 6S RNA candidates within the gamma-proteobacteria but were unable to identify reasonable 6S RNA candidates in other bacterial branches, utilizing mfold analyses of the region immediately upstream of ygfA combined with 6S RNA blastn searches. By RACE experiments, we mapped the major transcription initiation site of A.aeolicus 6S RNA primary transcripts, located within the pheT gene preceding ygfA, as well as three processing sites. PMID- 15814813 TI - Site-specific incorporation of N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (dG AAF) into oligonucleotides using modified 'ultra-mild' DNA synthesis. AB - Aromatic amino and nitro compounds are potent carcinogens found in the environment that exert their toxic effects by reacting with DNA following metabolic activation. One important adduct is N-(deoxyguanosin-8-yl)-2 acetylaminofluorene (dG-AAF), which has been extensively used in studies of the mechanisms of DNA repair and mutagenesis. Despite the importance of dG-AAF adducts in DNA, an efficient method for its incorporation into DNA using solid phase synthesis is still missing. We report the development of a modified 'ultra mild' DNA synthesis protocol that allows the incorporation of dG-AAF into oligonucleotides of any length accessible by solid-phase DNA synthesis with high efficiency and independent of sequence context. Key to this endeavor was the development of improved deprotection conditions (10% diisopropylamine in methanol supplemented with 0.25 M of beta-mercaptoethanol) designed to remove protecting groups of commercially available 'ultra-mild' phosphoramidite building blocks without compromising the integrity of the exquisitely base-labile acetyl group at N8 of dG-AAF. We demonstrate the suitability of these oligonucleotides in the nucleotide excision repair reaction. Our synthetic approach should facilitate comprehensive studies of the mechanisms of repair and mutagenesis induced by dG AAF adducts in DNA and should be of general use for the incorporation of base labile functionalities into DNA. PMID- 15814814 TI - Pre-organized structure of viral DNA at the binding-processing site of HIV-1 integrase. AB - The integration of the human immunodeficiency virus type 1 DNA into the host cell genome is catalysed by the viral integrase (IN). The reaction consists of a 3' processing [dinucleotide released from each 3' end of the viral long terminal repeat (LTR)] followed by a strand transfer (insertion of the viral genome into the human chromosome). A 17 base pair oligonucleotide d(GGAAAATCTCTAGCAGT), d(ACTGCTAGAGATTTTCC) reproducing the U5-LTR extremity of viral DNA that contains the IN attachment site was analysed by NMR using the classical NOEs and scalar coupling constants in conjunction with a small set of residual dipolar coupling constants (RDCs) measured at the 13C/15N natural abundance. The combination of these two types of parameters in calculations significantly improved the DNA structure determination. The well-known features of A-tracts were clearly identified by RDCs in the first part of the molecule. The binding/cleavage site at the viral DNA end is distinguishable by a loss of regular base stacking and a distorted minor groove that can aid its specific recognition by IN. PMID- 15814815 TI - Cyclic changes in the affinity of protein-DNA interactions drive the progression and regulate the outcome of the Tn10 transposition reaction. AB - The Tn10 transpososome is a DNA processing machine in which two transposon ends, a transposase dimer and the host protein integration host factor (IHF), are united in an asymmetrical complex. The transitions that occur during one transposition cycle are not limited to chemical cleavage events at the transposon ends, but also involve a reorganization of the protein and DNA components. Here, we demonstrate multiple pathways for Tn10 transposition. We show that one series of events is favored over all others and involves cyclic changes in the affinity of IHF for its binding site. During transpososome assembly, IHF is bound with high affinity. However, the affinity for IHF drops dramatically after cleavage of the first transposon end, leading to IHF ejection and unfolding of the complex. The ejection of IHF promotes cleavage of the second end, which is followed by restoration of the high affinity state which in turn regulates target interactions. PMID- 15814816 TI - Functional roles of 3'-terminal structures of template RNA during in vivo retrotransposition of non-LTR retrotransposon, R1Bm. AB - R1Bm is a non-LTR retrotransposon found specifically within 28S rRNA genes of the silkworm. Different from other non-LTR retrotransposons encoding two open reading frames (ORFs), R1Bm structurally lacks a poly (A) tract at its 3' end. To study how R1Bm initiates reverse transcription from the poly (A)-less template RNA, we established an in vivo retrotransposition system using recombinant baculovirus, and characterized retrotransposition activities of R1Bm. Target-primed reverse transcription (TPRT) of R1Bm occurred from the cleavage site generated by endonuclease (EN). The 147 bp of 3'-untranslated region (3'UTR) was essential for efficient retrotransposition of R1Bm. Even using the complete R1Bm element, however, reverse transcription started from various sites of the template RNA mostly with 5'-UG-3' or 5'-UGU-3' at their 3' ends, which are presumably base paired with 3' end of the EN-digested 28S rDNA target sequence, 5'-AGTAGATAGGGACA 3'. When the downstream sequence of 28S rDNA target was added to the 3' end of R1 unit, reverse transcription started exactly from the 3' end of 3'UTR and retrotransposition efficiency increased. These results indicate that 3'-terminal structure of template RNA including read-through region interacts with its target rDNA sequences of R1Bm, which plays important roles in initial process of TPRT in vivo. PMID- 15814817 TI - A novel cGUUAg tetraloop structure with a conserved yYNMGg-type backbone conformation from cloverleaf 1 of bovine enterovirus 1 RNA. AB - The 5'-terminal cloverleaf (CL)-like RNA structures are essential for the initiation of positive- and negative-strand RNA synthesis of entero- and rhinoviruses. SLD is the cognate RNA ligand of the viral proteinase 3C (3C(pro)), which is an indispensable component of the viral replication initiation complex. The structure of an 18mer RNA representing the apical stem and the cGUUAg D-loop of SLD from the first 5'-CL of BEV1 was determined in solution to a root-mean square deviation (r.m.s.d.) (all heavy atoms) of 0.59 A (PDB 1Z30). The first (antiG) and last (synA) nucleotide of the D-loop forms a novel 'pseudo base pair' without direct hydrogen bonds. The backbone conformation and the base-stacking pattern of the cGUUAg-loop, however, are highly similar to that of the coxsackieviral uCACGg D-loop (PDB 1RFR) and of the stable cUUCGg tetraloop (PDB 1F7Y) but surprisingly dissimilar to the structure of a cGUAAg stable tetraloop (PDB 1MSY), even though the cGUUAg BEV D-loop and the cGUAAg tetraloop differ by 1 nt only. Together with the presented binding data, these findings provide independent experimental evidence for our model [O. Ohlenschlager, J. Wohnert, E. Bucci, S. Seitz, S. Hafner, R. Ramachandran, R. Zell and M. Gorlach (2004) Structure, 12, 237-248] that the proteinase 3C(pro) recognizes structure rather than sequence. PMID- 15814818 TI - Dementia of the Alzheimer's type and accelerated aging in Down syndrome. AB - This case study, of a woman with Down syndrome and dementia of the Alzheimer's type (DAT), follows the course of her decline over an 11-year period until death at age 57. Detailed neuropathological findings are also presented. This case illustrates features of premature aging that are typically associated with Down syndrome, and the progressive changes in memory and cognition that are usually associated with DAT. Although the subject's cardiovascular condition and thyroid disorder were treated, they may have contributed to the decline of her memory. This case shows the difficulty in diagnosing dementia in an individual with mental retardation who suffered comorbid episodes of depression and psychosis. PMID- 15814819 TI - Power cut. PMID- 15814820 TI - Giant steps. PMID- 15814821 TI - Future mortality: a bumpy road to Shangri-La? AB - Americans are getting fatter, and it is known that increased obesity may increase the risk of death. Olshansky et al. have argued that this increase in obesity will likely slow, or even reverse, increases in life expectancy in the United States and perhaps save U.S. Social Security as a result. We discuss historical changes in the mortality rate and the reasons why other analyses argue that life expectancies will continue to increase. We also discuss the limitations of using single risk factors such as obesity as predictors of mortality risk. Finally, we explore the relation between risk factors and the long-term historical increase in human life expectancy. PMID- 15814822 TI - Mammalian-like nonsialyl complex-type N-glycosylation of equine gonadotropins in Mimic insect cells. AB - Recombinant equine luteinizing hormone/chorionic gonadotropin (eLH/CG) was expressed in Mimic insect cells, that are commercial stably transformed Spodoptera frugiperda (Sf9) cells expressing five mammalian genes encoding glycosyltransferases involved in the synthesis of complex-type monosialylated N glycans. We previously showed that it exhibited no in vivo bioactivity although expressing full in vitro bioactivity, and it was suspected that this was because of insufficient sialylation of eLH/CG N-glycans. Lectin binding analyses were performed with recombinant dimeric eLH/CG or its alpha subunit, secreted in the serum-containing supernatant of infected Sf9 and Mimic cells. Two types of specific lectin affinity assays (blot analyses and enzyme-linked immunosorbent assay) were used to compare the ability or inability of natural and recombinant gonadotropins to bind to various lectins. In natural equine chorionic gonadotropin (eCG), complex-type N-glycans terminating with both Siaalpha2,3Gal (based on Maackia amurensis agglutinin [MAA] binding) and Siaalpha2,6Gal (based on Sambucus nigra agglutinin [SNA] binding) were found, but in the alpha subunit dissociated from natural eCG, we only detected Siaalpha2-6Gal. In eLH/CG and its alpha subunit produced by Sf9 cells, N-glycans were found to be terminated by mannosyl residues (based on Galanthus nivalis agglutinin [GNA] binding), whereas those produced in Mimic cells were terminated by galactoses (based on binding to Ricinus communis agglutinin I [RCA I] , but not to SNA or MAA). This is in agreement with the fact that the nucleotide donor substrate of sialic acid is not naturally synthesized in insect cells. On the basis of binding to Arachis Hypogaea agglutinin [PNA], O-glycans exhibited the Galbeta1-3GalNAc structure in recombinant-free alpha and eLH/CG from both Sf9 and Mimic cell lines. Both N- and O-linked carbohydrate side chains synthesized in Mimic cells should thus be amenable to further acellular sialylation. PMID- 15814823 TI - MUC5B glycosylation in human saliva reflects blood group and secretor status. AB - This study aimed to characterize human salivary glycoforms and the natural glycosylation variation of the major ABO blood group bearing high molecular weight glycoprotein fraction MG1, which mainly consists of MUC5B mucin. Reduced and alkylated mucins from individuals of blood group A, B, and O were purified by sodium dodecyl sulfate-agarose/polyacrylamide composite gel electrophoresis (SDS AgPAGE), blotted to polyvinylidene fluoride (PVDF) membranes, and visualized with alcian blue. O-linked oligosaccharides were released from MUC5B glycoform bands by reductive beta-elimination and analyzed by liquid chromatography (LC) electrospray ion trap mass spectrometry (MS). Slow electrophoretically migrating MUC5B components (sm) were found to be dominated by neutral oligosaccharides, and fast-migrating (fm) components were dominated by sulfated oligosaccharides. ABO blood group-specific sequences were found on all glycoforms, and novel oligosaccharides containing blood group A and B type sequences were sequenced. This is the first molecular description of the influence of the blood group ABO system on salivary MUC5B oligosaccharides. Expanding these results from the three A, B, and O individuals into larger population (29 individuals), we found oligosaccharide sequences corresponding to the blood group of the donor on MUC5B from 23 individuals. The remaining six individuals were characterized by a high degree of sialylation. These individuals were assigned as nonsecretors, whereas blood group-expressing individuals were assigned as secretors. Western blot assays with antibodies confirmed increased expression of Sialyl Lewis a (Si Le(a)) in the nonsecretors. Our results highlight that salivary MUC5B consists of glycoforms with distinct glycosylation that vary extensively between individuals and that some of this variation is owing to blood group and secretor status. PMID- 15814824 TI - Sequence-variant repeats of MUC1 show higher conformational flexibility, are less densely O-glycosylated and induce differential B lymphocyte responses. AB - The human epithelial cancer mucin MUC1 is able to break tolerance and to induce humoral immune responses in healthy subjects and in cancer patients. We recently showed that clusters of sequence-variant repeats are interspersed in the repeat domain of MUC1 at high frequency, which should contribute to the structural and immunological features of the mucin. Here we elucidated the potential effects exerted by sequence-variant repeats on their O-glycosylation. Evidence from in vitro glycosylation with polypeptide N-acetylgalactosaminyltransferases GalNAc-T1 and GalNAc-T2 in concert with mass spectrometric analyses of in vivo glycosylated MUC1 probes from transiently transfected HEK293 cells indicated reduced glycosylation densities of repeats with three concerted replacements: AHGVTSAPESRPAPGSTAPA. The Pro to Ala replacement in STAPA exerts not only proximal effects on the ppGalNAc-T2 preferred site at -3 and -4, but also more distant effects on the ppGalNAc-T1 preferred site at -15 (TSAPESRPAPGSTAPA). We also examined the conformational changes of MUC1 glycopeptides induced by the concerted DT to ES replacements and revealed a higher conformational flexibility of ES/P peptides compared to DT/P peptides. Differences in conformational flexibilities and in O-glycosylation densities could underlie the observed differential humoral responses in humans. We were able to show that the natural immunoglobulin G (IgG) responses to the repeat domain of MUC1 in sera from nonmalignant control subjects are preferentially directed to variant repeat clusters. In contrast, the IgG response in patients with adenocarcinoma shifted to higher frequencies of preferential DTR peptide binding. PMID- 15814825 TI - Novel biosynthetic functions of lipopolysaccharide rfaJ homologs from Helicobacter pylori. AB - Activity screening and insertional inactivation of lipopolysaccharide (LPS) biosynthetic genes in Helicobacter pylori have led to the successful characterization of two key enzymes encoded by HP0159 (JHP0147) and HP1105 (JHP1032) open reading frames (ORFs) which are members of the large and diverse carbohydrate active enzymes (CAZY) GT-8 (rfaJ) family of glycosyltransferases. Activity screening of a genomic library led to the identification of the enzyme involved in the biosynthesis of the type 2 N-acetyl-lactosamine O-chain backbone, the beta-1,3-N-acetyl-glucosaminyl transferase. In addition, the activity screening approach led to the identification and characterization of a key core biosynthetic enzyme responsible for the biosynthesis of the alpha-1,6-glucan polymer. This alpha-1,6-glucosyltransferase protein is encoded by the HP0159 ORF. Both enzymes play an integral part in the biosynthesis of LPS, and insertional inactivation leads to the production of a truncated LPS molecule on the bacterial cell surface. The LPS structures were determined by mass spectrometry and chemical analyses. The linkage specificity of each glycosyltransferase was determined by nuclear magnetic resonance (NMR) analysis of model compounds synthesized in vitro. A cryogenic probe was used to structurally characterize nanomole amounts of the product of the HP1105 (JHP1032) enzyme. In contrast to the HP0159 enzyme, which displays the GT-8-predicted retaining stereochemistry for the reaction product, HP1105 (JHP1032) is the first member of this GT-8 family to have been shown to have an inverting stereochemistry in its reaction products. PMID- 15814826 TI - Time dependency of molecular rate estimates and systematic overestimation of recent divergence times. AB - Studies of molecular evolutionary rates have yielded a wide range of rate estimates for various genes and taxa. Recent studies based on population-level and pedigree data have produced remarkably high estimates of mutation rate, which strongly contrast with substitution rates inferred in phylogenetic (species level) studies. Using Bayesian analysis with a relaxed-clock model, we estimated rates for three groups of mitochondrial data: avian protein-coding genes, primate protein-coding genes, and primate d-loop sequences. In all three cases, we found a measurable transition between the high, short-term (< 1-2 Myr) mutation rate and the low, long-term substitution rate. The relationship between the age of the calibration and the rate of change can be described by a vertically translated exponential decay curve, which may be used for correcting molecular date estimates. The phylogenetic substitution rates in mitochondria are approximately 0.5% per million years for avian protein-coding sequences and 1.5% per million years for primate protein-coding and d-loop sequences. Further analyses showed that purifying selection offers the most convincing explanation for the observed relationship between the estimated rate and the depth of the calibration. We rule out the possibility that it is a spurious result arising from sequence errors, and find it unlikely that the apparent decline in rates over time is caused by mutational saturation. Using a rate curve estimated from the d-loop data, several dates for last common ancestors were calculated: modern humans and Neandertals (354 ka; 222-705 ka), Neandertals (108 ka; 70-156 ka), and modern humans (76 ka; 47-110 ka). If the rate curve for a particular taxonomic group can be accurately estimated, it can be a useful tool for correcting divergence date estimates by taking the rate decay into account. Our results show that it is invalid to extrapolate molecular rates of change across different evolutionary timescales, which has important consequences for studies of populations, domestication, conservation genetics, and human evolution. PMID- 15814827 TI - Dispersal and evolution of the Sinorhizobium meliloti group II RmInt1 intron in bacteria that interact with plants. AB - Group II introns are both self-splicing RNAs and mobile retroelements found in bacterial and archaeal genomes and in organelles of eukaryotes. They are thought to be the ancestors of eukaryote spliceosomal introns and non-long terminal repeat retrotransposons. We show here that RmInt1, a bacterial group II intron first described in the nitrogen-fixing symbiont of alfalfa (Medicago sativa) Sinorhizobium meliloti, is also present in other Sinorhizobium and Rhizobium species. The intron-homing sites in these species are IS elements of the ISRm2011 2 group as in S. meliloti, but ectopic insertion is also observed. We present evidence that these related bacteria have acquired RmInt1 by vertical inheritance from a common ancestor and by independent horizontal transfer events. We also show that RmInt1 is mobile in related taxa of bacteria that interact with plants and tends to evolve toward an inactive form by fragmentation, with loss of the 3' terminus including the intron-encoded protein. Our results provide an overview of the evolution and dispersion of a bacterial group II intron. PMID- 15814828 TI - Expanding Southwest Pacific mitochondrial haplogroups P and Q. AB - Modern humans have occupied New Guinea and the nearby Bismarck and Solomon archipelagos of Island Melanesia for at least 40,000 years. Previous mitochondrial DNA (mtDNA) studies indicated that two common lineages in this region, haplogroups P and Q, were particularly diverse, with the coalescence for P considered significantly older than that for Q. In this study, we expand the definition of haplogroup Q so that it includes three major branches, each separated by multiple mutational distinctions (Q1, equivalent to the earlier definition of Q, plus Q2 and Q3). We report three whole-mtDNA genomes that establish Q2 as a major Q branch. In addition, we describe 314 control region sequences that belong to the expanded haplogroups P and Q from our Southwest Pacific collection. The coalescence dates for the largest P and Q branches (P1 and Q1) are similar to each other (approximately 50,000 years old) and considerably older than prior estimates. Newly identified Q2, which was found in Island Melanesian samples just to the east, is somewhat younger by more than 10,000 years. Our coalescence estimates should be more reliable than prior ones because they were based on significantly larger samples as well as complete mtDNA coding region sequencing. Our estimates are roughly in accord with the current suggested dates for the first settlement of New Guinea-Sahul. The phylogeography of P and Q indicates almost total (female) isolation of ancient New Guinea-Island Melanesia from Australia that may have existed from the time of the first settlement. While Q subsequently diversified extensively in New Guinea-Island Melanesia, it has not been found in Australia. The only shared mtDNA haplogroup between Australia and New Guinea identified to date remains one minor branch of P. PMID- 15814829 TI - Understanding differences between phylogenetic and pedigree-derived mtDNA mutation rate: a model using families from the Azores Islands (Portugal). AB - We analyzed the control region of the mitochondrial DNA (mtDNA) from maternally related individuals originating from the Azores Islands (Portugal) in order to estimate the mutation rate of mtDNA and to gain insights into the process by which a new mutation arises and segregates into heteroplasmy. Length and/or point heteroplasmies were found at least in one individual of 72% of the studied families. Eleven new point substitutions were found, all of them in heteroplasmy, from which five appear to be somatic mutations and six can be considered germinal, evidencing the high frequency of somatic mutations in mtDNA in healthy young individuals. Different values of the mutation rate according to different assumptions were estimated. When considering all the germinal mutations, the value of the mutation rate obtained is one of the highest reported so far in family studies. However, when corrected for gender (assuming that the mutations present in men have the same evolutionary weight of somatic mutations because they will inevitably be lost) and for the probability of intraindividual fixation, the value for the mutation rate obtained for HVRI and HVRII (0.2415 mutations/site/Myr) was in the upper end of the values provided by phylogenetic estimations. These results indicate that the discrepancy, that has been reported previously, between the human mtDNA mutation rates observed along evolutionary timescales and the estimations obtained using family pedigrees can be minimized when corrections for gender proportions in newborn individuals and for the probability of intraindividual fixation are introduced. The analyses performed support the hypothesis that (1) in a constant, tight bottleneck genetic drift alone can explain different patterns of heteroplasmy segregation and (2) in neutral conditions, the destiny of a new mutation is strictly related to the initial proportion of the new variant. Another important point arising from the data obtained is that, even in the absence of a paternal contribution of mtDNA, recombination may occur between mtDNA molecules present in an individual, which is only observable if it occurs between mtDNA types that differ at two or more positions. PMID- 15814830 TI - Evolutionary history of 4.5SH RNA. AB - 4.5SH RNA is a 94-nt small RNA with unknown function. This RNA is known to be present in the mouse, rat, and hamster cells; however, it is not found in human, rabbit, and chicken. In the mouse genome, the 4.5SH RNA gene is a part of a long (4.2 kb) tandem repeat ( approximately 800 copies) unit. Here, we found that 4.5SH RNA genes are present only in rodents of six families that comprise the Myodonta clade: Muridae, Cricetidae, Spalacidae, Rhizomyidae, Zapodidae, and Dipodidae. The analysis of complementary DNA derived from the rodents of these families showed general evolutionary conservation of 4.5SH RNA and some intraspecific heterogeneity of these RNA molecules. 4.5SH RNA genes in the Norway rat, mole rat, hamster and jerboa genomes are included in the repeated sequences. In the jerboa genome these repeats are 4.0-kb long and arranged tandemly, similar to the corresponding arrangements in the mouse and rat genomic DNA. Sequencing of the rat and jerboa DNA repeats containing 4.5SH RNA genes showed fast evolution of the gene-flanking sequences. The repeat sequences of the distantly related rodents (mouse and rat vs. jerboa) have no apparent similarity except for the 4.5SH RNA gene itself. Conservation of the 4.5SH RNA gene nucleotide sequence indicates that this RNA is likely to be under selection pressure and, thus, may have a function. The repeats from the different rodents have similar lengths and contain many simple short repeats. The data obtained suggest that long insertions, deletions, and simple sequence amplifications significantly contribute in the evolution of the repeats containing 4.5SH RNA genes. The 4.5SH RNA gene seems to have originated 50-85 MYA in a Myodonta ancestor from a copy of the B1 short interspersed element. The amplification of the gene with the flanking sequences could result from the supposed cellular requirement of the intensive synthesis of 4.5SH RNA. Further Myodonta evolution led to dramatic changes of the repeat sequences in every lineage with the conservation of the 4.5SH RNA genes only. This gene, like some other relatively recently originated genes, could be a useful model for studying generation and evolution of non protein-coding genes. PMID- 15814831 TI - Low total vitamin C plasma level is a risk factor for cardiovascular morbidity and mortality in hemodialysis patients. AB - Hemodialysis patients are prone to deficiency of vitamin C, which constitutes the most abundant nonenzymatic antioxidant in blood. Because antioxidants are involved in the pathogenesis of atherosclerosis, the authors examined the association of total vitamin C plasma level with cardiovascular outcomes in such patients. One hundred thirty-eight consecutive maintenance hemodialysis patients (median age 61 yr, 90 males) were enrolled in a single-center study. At baseline, routine laboratory parameters were recorded, and predialysis total vitamin C plasma levels were measured by high-pressure liquid chromatography. Patients were prospectively followed-up for the occurrence of a primary composite endpoint consisting of fatal and nonfatal major adverse cardiovascular events (MACE) and for all-cause and cardiovascular mortality. MACE occurred in 35 patients (25%) over a period of median 30 mo, and 42 patients (30%) died [29 cardiovascular deaths (21% of total)]. Using Cox proportional hazards modeling, adjusted hazard ratios for the occurrence of MACE were 3.90 (95% confidence interval [CI]: 1.42 to 10.67; P = 0.008) and 3.03 (95% CI: 1.03 to 8.92; P = 0.044) for patients in the lower (<32 micromol/L) and middle (32 to 60 micromol/L) tertile of total vitamin C levels, compared with patients in the upper tertile (>60 micromol/L). Hazard ratios for cardiovascular death were 3.79 (95% CI: 1.23 to 11.66; P = 0.020) and 2.89 (95% CI: 0.89 to 9.37; P = 0.076). Total vitamin C levels were not independently associated with all-cause mortality. This study concludes that low total vitamin C plasma levels predict adverse cardiovascular outcomes among maintenance hemodialysis patients. Future studies should address the potential protective effect of an adequate vitamin C supplementation. PMID- 15814832 TI - Calcium, phosphorus, parathyroid hormone, and cardiovascular disease in hemodialysis patients: the USRDS waves 1, 3, and 4 study. AB - Animal studies suggest that calcium-phosphorus homeostatic abnormalities cause cardiovascular disease in uremia; few observational studies in humans have explored this. Associations in the retrospective United States Renal Data System Waves 1, 3, and 4 Study of 14,829 patients who were on hemodialysis on December 31, 1993, were examined. Mean age and duration of renal replacement therapy were 60.0 and 3.2 yr, respectively; 40.7% had diabetes. Quintiles (Q(1) to Q(5)) of (albumin-adjusted) calcium were 10.2 mg/dl; phosphorus, 7.5 mg/dl; calcium-phosphorus product, 71.0 mg(2)/dl(2); and parathyroid hormone (PTH), 480 pg/ml. Higher calcium levels were associated with fatal or nonfatal cardiovascular events (adjusted hazards ratio, 1.08 for Q(5), versus Q(1)) and all-cause mortality (Q(2), 1.07; Q(4), 1.11; Q(5), 1.14). Phosphorus levels were associated with cardiovascular events (Q(2), 1.06; Q(3), 1.13; Q(4), 1.14; Q(5), 1.25) and mortality (Q(4), 1.10; Q(5), 1.19), calcium phosphorus product was associated with cardiovascular events (Q(3), 1.09; Q(4), 1.14; Q(5), 1.24) and mortality (Q(4), 1.09; Q(5), 1.19), and PTH levels were associated with cardiovascular events (Q(5), 1.12) and mortality (Q(5), 1.17). Despite limitations (including retrospective design; noncurrent study era; and lack of serial calcium, phosphorus, and PTH measurements), this study suggests that disorders of calcium homeostasis are associated with fatal and nonfatal cardiovascular events and all-cause mortality in hemodialysis patients. PMID- 15814833 TI - And you thought the age of anatomic discovery was over. PMID- 15814834 TI - Molecular identification of canine podocalyxin-like protein 1 as a renal tubulogenic regulator. AB - GP135 is an apical membrane protein expressed in polarized MDCK epithelial cells. When cultured in three-dimensional collagen gel, MDCK cells form branching tubules in response to hepatocyte growth factor stimulation in a manner that simulates the embryonic renal development. During this process, GP135 displays transient loss of membranous localization but reappears at the cell surface when nascent lumen emerges from the developing tubules. Despite being used for decades as the canonical hallmark of apical surface, the molecular identity and the significance of the dynamic expression of GP135 during the tubulogenic process remain elusive. For exploring the function of GP135, the full-length cDNA encoding GP135 was obtained. Sequence alignments and features analysis confirm GP135 as a canine homolog of podocalyxin, confirming the finding of an earlier independent study. Immunohistochemical assays on canine kidney sections identified both glomerular and tubular distribution of GP135 along the nephron. Mutant MDCK cells expressing siRNA targeted at two regions of GP135 show defects in hepatocyte growth factor-induced tubulogenesis. Re-expression of full-length and an O-linked glycosylation abbreviated construct of GP135 could recapitulate the tubulogenesis process lacking in siRNA knockdown cells; however, a deletion construct devoid of the cytoplasmic domain failed to rescue the phenotype. In summary, the data identify the MDCK apical domain marker GP135 as a tubular form of podocalyxin and provide evidence for its importance in renal tubulogenesis. PMID- 15814835 TI - Proliferation of bone marrow-derived cells contributes to regeneration after folic acid-induced acute tubular injury. AB - Studies of tissue from recipients of bone marrow transplantation or organ allograft suggest that bone marrow-derived cells (BMDC) may differentiate into a variety of nonhematologic tissues, including renal tubular epithelium. The aims of this study were to examine whether BMDC contribute to recovery after acute renal injury and to assess the effects of cytokine mobilization on regeneration. Female mice (6 wk old) were lethally irradiated and transplanted with male bone marrow (BM) cells and later assigned into control, folic acid-treatment, and folic acid-treatment with granulocyte-colony stimulating factor (G-CSF), and control with G-CSF. Tritiated thymidine was given 1 h before death. Kidney sections were stained for a tubular epithelial marker, Y chromosome (in situ hybridization), periodic acid-Schiff staining, and subjected to autoradiography. Renal tubular epithelial cells in S-phase were scored as female (indigenous) or male (BM-derived). This is the first report to show that BMDC can respond by engrafting the renal tubules and undergo DNA synthesis after acute renal injury. BMDC contributed to the renal tubular epithelial cell population, although most (90%) renal tubular regeneration came from female indigenous cells. Some evidence was found for cell fusion between indigenous renal tubular cells and BMDC, but this was infrequent and the significance and consequences of cell fusion in the kidney are unresolved. G-CSF treatment nearly doubled the frequency of thymidine labeled BM-derived tubular cells and might facilitate the recovery of renal tubular epithelium. PMID- 15814836 TI - Nasal administration of recombinant rat alpha3(IV)NC1 prevents the development of experimental autoimmune glomerulonephritis in the WKY rat. AB - Experimental autoimmune glomerulonephritis (EAG), an animal model of Goodpasture's disease, can be induced in Wistar Kyoto (WKY) rats by immunization with either collagenase-solubilized rat glomerular basement membrane (GBM) or the recombinant NC1 domain of the alpha3 chain of type IV collagen [alpha3(IV)NC1]. EAG is characterized by circulating and deposited anti-glomerular basement membrane antibodies, focal necrotizing glomerulonephritis with crescent formation, and glomerular infiltration by T cells and macrophages. Previous studies have demonstrated that oral administration of collagenase-solubilized GBM to WKY rats prevented the development of EAG. Nasal administration of specific autoantigens has been reported to be more effective than oral administration in other models of autoimmune disease. The main aim of this study was to investigate further the concept of mucosal tolerance in EAG by examining the effect of nasal administration of recombinant rat alpha3(IV)NC1. Groups of WKY rats with EAG, induced by immunization with recombinant rat alpha3(IV)NC1, were given alpha3(IV)NC1 nasally on 3 consecutive days before immunization, at total cumulative doses of 25, 100, or 250 microg per rat. A dose-dependent effect was observed on the development of EAG. A dose of 25 microg had no effect on disease; 100 microg resulted in a moderate reduction in the severity of nephritis; and 250 microg led to a marked reduction in circulating and deposited antibodies, albuminuria, severity of glomerular abnormalities, and numbers of glomerular CD8+ T cells and macrophages. In addition, there was a reduction in the proliferative response of splenocytes from rats in the high dose group (250 microg) to alpha3(IV)NC1 in vitro. The results from this study clearly demonstrate for the first time that mucosal tolerance in EAG can be induced by nasal administration of recombinant rat alpha3(IV)NC1 and that this approach is effective in the prevention of crescentic glomerulonephritis. Further work using new antigen specific treatment strategies may provide a novel approach to the treatment of patients with anti-glomerular basement membrane disease. PMID- 15814837 TI - Akt mediates mechanical strain-induced collagen production by mesangial cells. AB - Increased glomerular hydrostatic pressure is an important determinant of glomerulosclerosis and can be modeled by in vitro exposure of mesangial cells to cyclic mechanical strain. Stretched mesangial cells increase extracellular matrix protein production, the hallmark of glomerulosclerosis. Recent data indicate that the serine/threonine kinase Akt may be involved in matrix modulation. Thus, Akt activation and matrix synthesis in stretched mesangial cells were studied. Exposure of mesangial cells to 1 Hz cyclic strain led to prompt Akt activation, which was biphasic to 24 h. Activation was dependent on signaling through phosphatidylinositol-3-kinase and required EGF receptor transactivation. Inhibition of signaling through the PDGF receptor, Src kinase, or cytoskeletal disruption failed to prevent strain-induced Akt activation. Collagen type 1A1 transcript expression, promoter activation, and protein secretion were increased by stretch at 24 h and were dependent on phosphatidylinositol-3 kinase. Overexpression of dominant-negative Akt inhibited strain-induced collagen 1A1 production. Conversely, overexpression of constitutively active Akt led to increased collagen 1A1 upregulation and secretion. Finally, Akt activation was observed in the glomeruli of remnant rat kidneys, a model marked by increased intraglomerular pressure. The authors conclude that mechanical strain induces Akt activation in mesangial cells through a mechanism requiring phosphatidylinositol 3-kinase and EGF receptor transactivation. Type 1 collagen production is dependent on Akt and can be induced by Akt overexpression. Akt activation is observed in remnant kidneys in vivo. Thus, the role of Akt in progression of chronic hemodynamic glomerular disease is worthy of further exploration. PMID- 15814838 TI - Nuclear targeting of adenovirus type 2 requires CRM1-mediated nuclear export. AB - Incoming adenovirus type 2 (Ad2) and Ad5 shuttle bidirectionally along microtubules, biased to the microtubule-organizing center by the dynein/dynactin motor complex. It is unknown how the particles reach the nuclear pore complex, where capsids disassemble and viral DNA enters the nucleus. Here, we identified a novel link between nuclear export and microtubule-mediated transport. Two distinct inhibitors of the nuclear export factor CRM1, leptomycin B (LMB) and ratjadone A (RJA) or CRM1-siRNAs blocked adenovirus infection, arrested cytoplasmic transport of viral particles at the microtubule-organizing center or in the cytoplasm and prevented capsid disassembly and nuclear import of the viral genome. In mitotic cells where CRM1 is in the cytoplasm, adenovirus particles were not associated with microtubules but upon LMB treatment, they enriched at the spindle poles implying that CRM1 inhibited microtubule association of adenovirus. We propose that CRM1, a nuclear factor exported by CRM1 or a protein complex containing CRM1 is part of a sensor mechanism triggering the unloading of the incoming adenovirus particles from microtubules proximal to the nucleus of interphase cells. PMID- 15814839 TI - Role for the SCFCDC4 ubiquitin ligase in Candida albicans morphogenesis. AB - The ability of Candida albicans, a major fungal pathogen, to switch between a yeast form, and a hyphal (mold) form is recognized as being important for the ability of the organism to invade the host and cause disease. We found that a C. albicans mutant deleted for CaCDC4, a homologue of the Saccharomyces cerevisiae F box protein component of the SCF(CDC4) ubiquitin ligase, is viable and displays constitutive filamentous, mostly hyphal, growth. The phenotype of the Cacdc4-/- mutant suggests that ubiquitin-mediated protein degradation is involved in the regulation of the dimorphic switch of C. albicans and that one or more regulators of the yeast-to-mold switch are among the substrates of SCF(CaCDC4). Epistasis analysis indicates that the Cacdc4-/- phenotype is largely independent of the filamentation-inducing transcription factors Efg1 and Cph1. We identify C. albicans Far1 and Sol1, homologues of the S. cerevisiae SCF(CDC4) substrates Far1 and Sic1, and show that Sol1 is a substrate of C. albicans Cdc4. Neither protein is essential for the hyphal phenotype of the Cacdc4-/- mutant. However, ectopic expression and deletion of SOL1 indicate a role for this gene in C. albicans morphogenesis. PMID- 15814840 TI - Induction of the Candida albicans filamentous growth program by relief of transcriptional repression: a genome-wide analysis. AB - Candida albicans, the major human fungal pathogen, undergoes a reversible morphological transition from blastospores (round budding cells) to filaments (elongated cells attached end-to-end). This transition, which is induced upon exposure of C. albicans cells to a number of host conditions, including serum and body temperature (37 degrees C), is required for virulence. Using whole-genome DNA microarray analysis, we describe 61 genes that are significantly induced (> or =2-fold) during the blastospore to filament transition that takes place in response to exposure to serum and 37 degrees C. We next show that approximately half of these genes are transcriptionally repressed in the blastospore state by three transcriptional repressors, Rfg1, Nrg1, and Tup1. We conclude that the relief of this transcriptional repression plays a key role in bringing the C. albicans filamentous growth program into play, and we describe the framework of this transcriptional circuit. PMID- 15814841 TI - Global roles of Ssn6 in Tup1- and Nrg1-dependent gene regulation in the fungal pathogen, Candida albicans. AB - In budding yeast, Tup1 and Ssn6/Cyc8 form a corepressor that regulates a large number of genes. This Tup1-Ssn6 corepressor appears to be conserved from yeast to man. In the pathogenic fungus Candida albicans, Tup1 regulates cellular morphogenesis, phenotypic switching, and metabolism, but the role of Ssn6 remains unclear. We show that there are clear differences in the morphological and invasive phenotypes of C. albicans ssn6 and tup1 mutants. Unlike Tup1, Ssn6 depletion promoted morphological events reminiscent of phenotypic switching rather than filamentous growth. Transcript profiling revealed minimal overlap between the Ssn6 and Tup1 regulons. Hypha-specific genes, which are repressed by Tup1 and Nrg1, were not derepressed in ssn6 cells under the conditions studied. In contrast, the phase specific gene WH11 was derepressed in ssn6 cells, but not in tup1 or nrg1 cells. Hence Ssn6 and Tup1 play distinct roles in C. albicans. Nevertheless, both Ssn6 and Tup1 were required for the Nrg1-mediated repression of an artificial NRE promoter, and lexA-Nrg1 mediated repression in the C. albicans one-hybrid system. These observations are explained in models that are generally consistent with the Tup1-Ssn6 paradigm in budding yeast. PMID- 15814842 TI - The ATP-dependent membrane localization of protein kinase Calpha is regulated by Ca2+ influx and phosphatidylinositol 4,5-bisphosphate in differentiated PC12 cells. AB - Signal transduction through protein kinase Cs (PKCs) strongly depends on their subcellular localization. Here, we investigate the molecular determinants of PKCalpha localization by using a model system of neural growth factor (NGF) differentiated pheochromocytoma (PC12) cells and extracellular stimulation with ATP. Strikingly, the Ca2+ influx, initiated by the ATP stimulation of P2X receptors, rather than the Ca2+ released from the intracellular stores, was the driving force behind the translocation of PKCalpha to the plasma membrane. Furthermore, the localization process depended on two regions of the C2 domain: the Ca2+-binding region and the lysine-rich cluster, which bind Ca2+ and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2], respectively. It was demonstrated that diacylglycerol was not involved in the localization of PKCalpha through its C1 domain, and in lieu, the presence of PtdIns(4,5)P2 increased the permanence of PKCalpha in the plasma membrane. Finally, it also was shown that ATP cooperated with NGF during the differentiation process of PC12 cells by increasing the length of the neurites, an effect that was inhibited when the cells were incubated in the presence of a specific inhibitor of PKCalpha, suggesting a possible role for this isoenzyme in the neural differentiation process. Overall, these results show a novel mechanism of PKCalpha activation in differentiated PC12 cells, where Ca2+ influx, together with the endogenous PtdIns(4,5)P2, anchor PKCalpha to the plasma membrane through two distinct motifs of its C2 domain, leading to enzyme activation. PMID- 15814843 TI - Tracking the interactions of rRNA processing proteins during nucleolar assembly in living cells. AB - Reorganization of the nuclear machinery after mitosis is a fundamental but poorly understood process. Here, we investigate the recruitment of the nucleolar processing proteins in the nucleolus of living cells at the time of nucleus formation. We question the role of the prenucleolar bodies (PNBs), during migration of the processing proteins from the chromosome periphery to sites of rDNA transcription. Surprisingly, early and late processing proteins pass through the same PNBs as demonstrated by rapid two-color four-dimensional imaging and quantification, whereas a different order of processing protein recruitment into nucleoli is supported by differential sorting. Protein interactions along the recruitment pathway were investigated using a promising time-lapse analysis of fluorescence resonance energy transfer. For the first time, it was possible to detect in living cells the interactions between proteins of the same rRNA processing machinery in nucleoli. Interestingly interactions between such proteins also occur in PNBs but not at the chromosome periphery. The dynamics of these interactions suggests that PNBs are preassembly platforms for rRNA processing complexes. PMID- 15814844 TI - Mature DIABLO/Smac is produced by the IMP protease complex on the mitochondrial inner membrane. AB - DIABLO/Smac is a mitochondrial protein that can promote apoptosis by promoting the release and activation of caspases. To do so, DIABLO/Smac must first be processed by a mitochondrial protease and then released into the cytosol, and we show this in an intact cellular system. We propose that the precursor form of DIABLO/Smac enters the mitochondria through a stop-transfer pathway and is processed to its active form by the inner membrane peptidase (IMP) complex. Catalytic subunits of the mammalian IMP complex were identified based on sequence conservation and functional complementation, and the novel sequence motif RX(5)P in Imp1 and NX(5)S in Imp2 distinguish the two catalytic subunits. DIABLO/Smac is one of only a few specific proteins identified as substrates for the IMP complex in the mitochondrial intermembrane space. PMID- 15814845 TI - The nongenomic actions of aldosterone. AB - Aldosterone has physiological effects to regulate fluid and electrolyte homeostasis across epithelia and proinflammatory effects on a variety of nonepithelial cells in the context of inappropriate salt status. These effects are mediated by mineralocorticoid receptors, members of a large family of nuclear transcription factors, by DNA-directed, RNA-mediated protein synthesis. Rapid effects of aldosterone, insensitive to actinomycin D or cycloheximide and thus clearly nongenomic, have been convincingly documented in a variety of epithelial and nonepithelial tissues. Despite strenuous attempts, isolation of a nonclassical membrane receptor for aldosterone has proven unsuccessful, and rapid nongenomic effects mediated by classical mineralocorticoid receptors are increasingly recognized in the kidney, heart, and vascular wall. The mechanism of rapid nongenomic actions of aldosterone may vary between tissues in terms of pathways; in addition, what remains to be established is the physiological role of aldosterone action via such rapid nongenomic mechanisms and how they might synergize with the longer time course genomic actions of mineralocorticoids. PMID- 15814846 TI - The search for safer glucocorticoid receptor ligands. AB - Steroidal glucocorticoids are commonly used due to their powerful antiinflammatory activity. However, despite their excellent efficacy, severe side effects frequently limit the use of these drugs. The search for novel glucocorticoids with reduced side effects has been intensified by the discovery of new molecular details regarding the function of the glucocorticoid receptor. These new insights may pave the way for novel, safer therapies that retain the efficacy of currently prescribed steroids. PMID- 15814847 TI - Role of aldose reductase and oxidative damage in diabetes and the consequent potential for therapeutic options. AB - Aldose reductase (AR) is widely expressed aldehyde-metabolizing enzyme. The reduction of glucose by the AR-catalyzed polyol pathway has been linked to the development of secondary diabetic complications. Although treatment with AR inhibitors has been shown to prevent tissue injury in animal models of diabetes, the clinical efficacy of these drugs remains to be established. Recent studies suggest that glucose may be an incidental substrate of AR, which appears to be more adept in catalyzing the reduction of a wide range of aldehydes generated from lipid peroxidation. Moreover, inhibition of the enzyme has been shown to increase inflammation-induced vascular oxidative stress and prevent myocardial protection associated with the late phase of ischemic preconditioning. On the basis of these studies, several investigators have ascribed an important antioxidant role to the enzyme. Additionally, ongoing work indicates that AR is a critical component of intracellular signaling, and inhibition of the enzyme prevents high glucose-, cytokine-, or growth factor-induced activation of protein kinase C and nuclear factor-kappa-binding protein. Thus, treatment with AR inhibitors prevents vascular smooth muscle cell growth and endothelial cell apoptosis in culture and inflammation and restenosis in vivo. Additional studies indicate that the antioxidant and signaling roles of AR are interlinked and that AR regulates protein kinase C and nuclear factor-kappaB via redox-sensitive mechanisms. These data underscore the need for reevaluating anti-AR interventions for the treatment of diabetic complications. Potentially, the development of newer drugs that selectively inhibit AR-mediated glucose metabolism and signaling, without affecting aldehyde detoxification, may be useful in preventing inflammation associated with the development of diabetic complications, particularly micro- and macrovascular diseases. PMID- 15814848 TI - Development of growth hormone secretagogues. AB - The GH secretagogues (GHS) were developed by reverse pharmacology. The objective was to develop small molecules with pharmacokinetics suitable for once-daily oral administration that would rejuvenate the GH/IGF-I axis. Neither the receptor nor the ligand that controlled pulse amplitude of hormone release was known; therefore, identification of lead structures was based on function. I reasoned that GH pulse amplitude could be increased by four possible mechanisms: 1) increasing GHRH release; 2) amplifying GHRH signaling in somatotrophs of the anterior pituitary gland; 3) reducing somatostatin release; and 4) antagonizing somatostatin receptor signaling. Remarkably, the GHS act through all four mechanisms to reproduce a young adult physiological GH profile in elderly subjects that was accompanied by increased bone mineral density and lean mass, modest improvements in strength, and improved recovery from hip fracture. Furthermore, restoration of thymic function was induced in old mice. The GHS receptor (GHS-R) was subsequently identified by expression cloning and found to be a previously unknown G protein-coupled receptor expressed predominantly in brain, pituitary gland, and pancreas. Reverse pharmacology was completed when the cloned GHS-R was exploited to identify an endogenous agonist (ghrelin) and a partial agonist (adenosine); ghsr-knockout mice studies confirmed that GHS are ghrelin mimetics. PMID- 15814849 TI - Transcriptional regulation by steroid receptor coactivator phosphorylation. AB - The basic mechanisms underlying ligand-dependent transcriptional activation by nuclear receptors (NRs) require the sequential recruitment of various coactivators. Increasing numbers of coactivators have been identified in recent years, and both biochemical and genetic studies demonstrate that these coactivators are differentially used by transcription factors, including NRs, in a cell/tissue type- and promoter-specific manner. However, the molecular basis underlying this specificity remains largely unknown. Recently, NRs and coregulators were shown to be targets of posttranslational modifications activated by diverse cellular signaling pathways. It is argued that posttranslational modifications of these proteins provide the basis for a combinatorial code required for specific gene activation by NRs and coactivators, and that this code also enables coactivators to efficiently stimulate the activity of other classes of transcription factors. In this review, we will focus on coactivators and discuss the recent progress in understanding the role of phosphorylation of the steroid receptor coactivator family and the potential ramifications of this posttranslational modification for regulation of gene expression. PMID- 15814850 TI - Development and potential clinical uses of human prolactin receptor antagonists. AB - There is a large body of literature showing that prolactin (PRL) exerts growth promoting activities in breast cancer, and possibly in prostate cancer and prostate hyperplasia. In addition, increasing evidence argues for the involvement of locally produced (autocrine) PRL, perhaps even more than pituitary-secreted (endocrine) PRL, in tumor growth. Because dopamine analogs are unable to inhibit PRL production in extrapituitary sites, alternative strategies need investigation. To that end, several PRL receptor antagonists have been developed by introducing various mutations into its natural ligands. For all but one of these analogs, the mechanism of action involves a competition with endogenous PRL for receptor binding. Such compounds are thus candidates to counteract the undesired actions of PRL, not only in tumors, but also in dopamine-resistant prolactinomas. In this review, we describe the different versions of antagonists that have been developed, with emphasis on the controversies regarding their characterization, and the limits for their potential development as a drug. The most recently developed antagonist, Delta1-9-G129R-hPRL, is the only one that is totally devoid of residual agonistic activity, meaning it acts as pure antagonist. We discuss to what extent this new molecule could be considered as a lead compound for inhibiting the actions of human PRL in the above-mentioned diseases. We also speculate on the multiple questions that could be addressed with respect to the therapeutic use of PRL receptor antagonists in patients. PMID- 15814851 TI - Aromatase inhibitors in the treatment of breast cancer. AB - Estradiol, the most potent endogenous estrogen, is biosynthesized from androgens by the cytochrome P450 enzyme complex called aromatase. Aromatase is present in breast tissue, and intratumoral aromatase is the source of local estrogen production in breast cancer tissues. Inhibition of aromatase is an important approach for reducing growth-stimulatory effects of estrogens in estrogen dependent breast cancer. Steroidal inhibitors that have been developed to date build upon the basic androstenedione nucleus and incorporate chemical substituents at varying positions on the steroid. Nonsteroidal aromatase inhibitors can be divided into three classes: aminoglutethimide-like molecules, imidazole/triazole derivatives, and flavonoid analogs. Mechanism-based aromatase inhibitors are steroidal inhibitors that mimic the substrate, are converted by the enzyme to a reactive intermediate, and result in the inactivation of aromatase. Both steroidal and nonsteroidal aromatase inhibitors have shown clinical efficacy in the treatment of breast cancer. The potent and selective third-generation aromatase inhibitors, anastrozole, letrozole, and exemestane, were introduced into the market as endocrine therapy in postmenopausal patients failing antiestrogen therapy alone or multiple hormonal therapies. These agents are currently approved as first-line therapy for the treatment of postmenopausal women with metastatic estrogen-dependent breast cancer. Several clinical studies of aromatase inhibitors are currently focusing on the use of these agents in the adjuvant setting for the treatment of early breast cancer. Use of an aromatase inhibitor as initial therapy or after treatment with tamoxifen is now recommended as adjuvant hormonal therapy for a postmenopausal woman with hormone-dependent breast cancer. PMID- 15814852 TI - All hype, no hope? Excessive pessimism in the "anti-aging medicine" special sections. PMID- 15814854 TI - Regaining lost youth: the controversial and colorful beginnings of hormone replacement therapy in aging. AB - The quest for regaining lost youth seems to have existed since the beginning of recorded history and has taken many forms. One strategy that began in earnest in the latter part of the 19th century and continues to have enormous momentum today is based on the notion that by replacing internally secreted substances, that is, hormones, that decline with age, the vitality and physical attributes associated with youth can be regained. Although the approach remains highly controversial as, for example, in "anti-aging medicine," it is no more controversial than it was many years ago when the work of three high profile investigators, Charles Eduoard Brown-Sequard, Eugen Steinach, and Serge Voronoff set the basis for using this strategy. In the case of all three individuals, the therapies they developed received widespread attention (including ridicule) in the popular press, were spread rapidly by practitioners of questionable training and ethical motivation, and finally and relatively quickly disappeared from common use. However, and ultimately more importantly, in the process of developing and promoting their therapies, these individuals made important contributions to the origins of endocrinology, the biology of sex, and establishment of hormone replacement therapy. It remains to be seen whether contemporary efforts using hormone replacement therapy to blunt and reverse aging have the same fate as their predecessors and make comparable important contributions to biology and medicine. PMID- 15814855 TI - Differences in size, strength, and power of upper and lower body muscle groups in young and older men. AB - We compared muscle thickness, torque, normalized torque (torque/muscle thickness), and power at 1.05 rad/s and 3.14 rad/s in flexor and extensor muscles of the elbow and knee, and in ankle plantar flexors in young (n=22, 18-31 years) and older (n=28, 59-76 years) men. Young men had greater muscle thickness for all muscle groups (p<.01), except elbow extensors, which were similar to older men. Young men had greater torque and power at both velocities for all muscle groups (p<.01), and greater normalized torque at both velocities for the elbow extensors and knee flexors and at the fast velocity for knee extensors. Relative to young mean values, muscle thickness, and torque, normalized torque, and power in the older group were most affected for lower-body measurements, especially at the fast velocity. Torque, normalized torque, and power (especially at fast velocities), and muscle thickness in the lower body are affected more by aging than are upper body measures in men. PMID- 15814856 TI - Brief food restriction in old animals decreases triglyceride content and insulin stimulated triglyceride synthesis. AB - To determine the effects of brief food restriction on fatty acid (FA) metabolism in old muscle, hind limbs of 24-month F344/BN rats fed either ad libitum (AL) or 60% food restricted (FR) for 28 days were perfused under hyperglycemic hyperinsulinemic conditions. Basal glucose and insulin levels were significantly lower (p<.05) in FR rats. Although palmitate uptake was not affected by food restriction, palmitate oxidation was 49% lower (2.2+/-0.3 vs 4.3+/-0.7 nmol . g-1 . min-1, p<.05) in FR versus AL animals, respectively. Compared to AL animals, FR animals had 25%-43% (p<.05) lower muscle triglyceride (TG) levels and hyperinsulinemic TG synthesis rates. Higher glucose uptake rates occurred in FR rats (p<.05). In conclusion, our results indicate that brief food restriction in old animals improves insulin sensitivity as it pertains to both glucose uptake and FA oxidation. Together with the decrease in nonoxidative FA disposal, the decreased FA oxidation under hyperinsulinemic conditions may significantly contribute to food restriction-induced reduction in muscle TG. PMID- 15814857 TI - Selective atrophy of left hemisphere and frontal lobe of the brain in old men. AB - In this study, volumes of the whole brain, hemispheres, and frontal lobes of young and elderly adults were quantified by an automated method. Effects of age, sex, and side on absolute and relative volumes of the brain structures were evaluated. Compared with the young group, elderly participants showed a 15% volume loss in the whole brain and hemispheres, and a 22% volume loss in the frontal lobes. The relative volume of the left hemisphere in the elderly group decreased more than that of the right hemisphere. Elderly men showed significantly greater left hemisphere and left frontal lobe volume losses than did elderly women, indicating that the larger left hemisphere relative volume reduction is largely contributed to by selective atrophy of the left frontal lobe volume in elderly men. These results may reflect age- and sex-related functional deterioration in the left brain. PMID- 15814858 TI - Lack of effect of caloric restriction on bioenergetics and reactive oxygen species production in intact rat hepatocytes. AB - To investigate the hypothesis that caloric restriction alters mitochondrial function in situ, intact hepatocytes were isolated from fully fed and calorie restricted (55% of control food intake, 4 months duration) male Brown-Norway rats at 6 months of age, and various parameters were determined. Overall, the production of reactive oxygen species was not affected by caloric restriction, neither were the mitochondrial membrane potential, oxygen consumption driving proton leak, or oxygen consumption driving ATP turnover. It is concluded that while isolated mitochondria from liver tissue of calorie-restricted animals display a reduction in the generation of reactive oxygen species, it was not possible to confirm this effect in isolated hepatocytes. Further work is required to establish what effect, if any, caloric restriction has on the rate of generation of reactive oxygen species in intact cells and tissues and importantly at the whole-animal level. PMID- 15814859 TI - The effect of meals at different mealtimes on blood pressure and symptoms in geriatric patients with postprandial hypotension. PMID- 15814861 TI - Tai Chi and fall reductions in older adults: a randomized controlled trial. AB - BACKGROUND: The authors' objective was to evaluate the efficacy of a 6-month Tai Chi intervention for decreasing the number of falls and the risk for falling in older persons. METHODS: This randomized controlled trial involved a sample of 256 physically inactive, community-dwelling adults aged 70 to 92 (mean age, 77.48 years; standard deviation, 4.95 years) who were recruited through a patient database in Portland, Oregon. Participants were randomized to participate in a three-times-per-week Tai Chi group or to a stretching control group for 6 months. The primary outcome measure was the number of falls; the secondary outcome measures included functional balance (Berg Balance Scale, Dynamic Gait Index, Functional Reach, and single-leg standing), physical performance (50-foot speed walk, Up&Go), and fear of falling, assessed at baseline, 3 months, 6 months (intervention termination), and at a 6-month postintervention follow-up. RESULTS: At the end of the 6-month intervention, significantly fewer falls (n=38 vs 73; p=.007), lower proportions of fallers (28% vs 46%; p=.01), and fewer injurious falls (7% vs 18%; p=.03) were observed in the Tai Chi group compared with the stretching control group. After adjusting for baseline covariates, the risk for multiple falls in the Tai Chi group was 55% lower than that of the stretching control group (risk ratio,.45; 95% confidence interval, 0.30 to 0.70). Compared with the stretching control participants, the Tai Chi participants showed significant improvements (p<.001) in all measures of functional balance, physical performance, and reduced fear of falling. Intervention gains in these measures were maintained at a 6-month postintervention follow-up in the Tai Chi group. CONCLUSIONS: A three-times-per-week, 6-month Tai Chi program is effective in decreasing the number of falls, the risk for falling, and the fear of falling, and it improves functional balance and physical performance in physically inactive persons aged 70 years or older. PMID- 15814862 TI - Total and differential white blood cell counts and their associations with circulating interleukin-6 levels in community-dwelling older women. AB - BACKGROUND: Interleukin-6 (IL-6) is an inflammatory biomediator, and age-related increases in IL-6 levels are associated with osteoporosis, sarcopenia, disability, and mortality in older adults. Although white blood cells (WBC), or leukocytes, are known to produce IL-6 in vitro, their in vivo relationship with circulating IL-6 levels is not well established. METHODS: In this cross-sectional analysis of data from the Women's Health and Aging Study I, the authors evaluated the relationships of total WBC and WBC differential (neutrophil, monocyte, lymphocyte, eosinophil, and basophil) counts to circulating IL-6 levels in 619 community-dwelling older women. Potential associations of age, race, and cigarette smoking with total and differential WBC counts and IL-6 levels were also assessed. RESULTS: Except for lymphocyte and basophil counts, significant associations of total WBC, neutrophil, monocyte, and eosinophil counts with IL-6 levels were identified. These associations remained highly significant after adjustment for age, race, and smoking status. Total WBC, neutrophil, monocyte, and eosinophil counts had significant stepwise increases in four escalating quartiles of IL-6 levels. In addition, age, race, and cigarette smoking were differentially associated with total and differential WBC counts and IL-6 levels. CONCLUSIONS: Positive in vivo associations of total WBC and its specific subpopulations were identified, including neutrophils, monocytes, and eosinophils, with circulating IL-6 levels in community-dwelling older women. These findings suggest significant contributions of WBC and its subpopulations to circulating IL-6 levels and potential effects from chronic elevation of IL-6 levels to the function of these circulating immune cells that warrant further investigation. PMID- 15814863 TI - Age-related changes in microvascular blood flow and transcutaneous oxygen tension under Basal and stimulated conditions. AB - BACKGROUND: Adequate cutaneous microvascular blood flow and tissue oxygen tension are important prerequisites for successful tissue repair. The efficacy of tissue repair decreases with age and is linked to the age-related functional decline of unmyelinated sensory neurons that are important for inflammation and tissue repair. However, available information on the effect of these neuronal changes on microvascular blood flow and tissue oxygen tension is limited, particularly under control and injury conditions. The authors had two aims in this study: (a) to assess age-related changes in the relationship between microvascular blood flow and tissue oxygen perfusion under basal and two different stimulated conditions (sensory dependent and sensory independent), and (b) to clarify the biological meaning of transcutaneous partial pressure of oxygen (tcPO2) measurements. METHODS: The effects of a sensory-independent vasodilator (acetylcholine) and a sensory-dependent vasodilator (capsaicin) on microvascular blood flow and oxygen perfusion in persons of different ages were measured. Laser Doppler flowmetry and a commercially available transcutaneous oxygen monitor (with sensors set at 39 degrees C and 44 degrees C) were used. Healthy volunteers were recruited: 11 young, 14 middle aged, and 19 older. RESULTS: Under basal conditions (skin temperature, 37 degrees C to 39 degrees C), both basal blood flow and tcPO2 increased with increasing age. However, with the sensor set at 44 degrees C, tcPO2 showed a significant decrease with age. Acetylcholine increased blood flow approximately equally in the three age groups. Capsaicin increased blood flow and tcPO2 in all age groups, with the young showing a greater increase compared with the older participants. CONCLUSIONS: The age-associated changes in basal and stimulated microvascular blood flow and tcPO2 could be attributed in part to altered neuronal function. Measuring tcPO2 at 39 degrees C showed a trend toward an increase with age. In contrast, a decrease with age was observed when tcPO2 was measured at 44 degrees C, a temperature sufficient to activate sensory nerve endings. The results may reflect a decline in sensory nerve function with age rather than a decrease in oxygen delivery for vascular reasons. This is supported by the complementary data showing a significant age-related decrease in stimulated blood flow in response to capsaicin, with no change in the response to the sensory-independent vasodilator acetylcholine. Thus, for clinical purposes, data obtained using the tcPO2 monitor should be interpreted with full knowledge of the conditions under which the measurements were made. Furthermore, for scientific purposes, the tcPO2 monitor could be used to assess sensory nerve function when sensors are heated to 44 degrees C. PMID- 15814864 TI - Inner city, middle-aged African Americans have excess frank and subclinical disability. AB - BACKGROUND: Healthy People 2010 seeks to decrease or eliminate the health disparities experienced by disadvantaged minority groups. METHODS: African American Health (AAH) is a population-based panel study of community-dwelling African Americans born between 1936 and 1950 from two strata. The first encompasses a poor, inner city area, and the second involves a suburban population with higher socioeconomic status. The authors recruited 998 participants (76% recruitment). Frank disability was assessed for 25 tasks and defined as inability or difficulty performing that task. Subclinical disability was assessed for 12 tasks and defined as no difficulty but a change in either manner or frequency of task performance. Frank disability prevalences were compared with national data for community-dwelling non-Hispanic white persons (NHW) and African American persons in the same age range. RESULTS: Compared with the suburban sample, the inner city group had a higher prevalence of frank disability for all 25 tasks (p<.05 for 16) and subclinical disability for 11 of the 12 tasks (p<.05 for 5). Both strata had more frank disability compared with the national NHW population. The inner city area had higher frank disability proportions than did the national African American sample, whereas the suburban group had similar disability levels. CONCLUSIONS: The AAH inner city group experiences more frank disability than other populations of African Americans and NHWs. The increased prevalence of subclinical disability in the inner city group compared with the suburban group suggests that the disparity in frank disability will continue. These findings indicate that African Americans living in poor inner city areas in particular need intensive and targeted clinical and public health efforts. PMID- 15814865 TI - Prevalence of and risk factors for sarcopenia in elderly Chinese men and women. AB - BACKGROUND: Several studies have documented the substantial health and economic burdens associated with sarcopenia among the elderly, but there has been no systematic study among Asians. A cross-sectional survey of elderly community dwelling Chinese volunteers (262 men and 265 women), aged 70 years and older, was undertaken in Hong Kong. The purposes of this study were to evaluate the prevalence of and risk factors for sarcopenia in elderly Chinese, and to compare these observations with those in white persons. METHODS: Muscle mass was estimated by dual-energy X-ray absorptiometry. In this study, sarcopenia was defined as a total adjusted skeletal muscle mass two standard deviations or more below the normal mean for young Asian men and women in this study. The relationship between risk factors (alcohol consumption, cigarette smoking, regular exercise, body mass index, medical conditions) and sarcopenia was studied by multiple logistic regression. RESULTS: The prevalence of sarcopenia was 12.3% in Chinese men and 7.6% in Chinese women aged 70 years and older, which was slightly lower than figures observed in white persons. Being underweight was a significant risk factor in both men (odds ratio, 39.1; 95% confidence interval, 11.3 to 134.6) and women (odds ratio, 9.7; 95% confidence interval, 2.8 to 33.8). No other risk factors were found in Chinese men or women. CONCLUSIONS: Sarcopenia exists among elderly Chinese men and women, albeit at a lower rate than in white persons. This may be due to the lower muscle mass among young men and women or to an attenuated rate of loss in muscle mass with aging in the Chinese elderly. Being underweight is a major risk factor for sarcopenia in both sexes. PMID- 15814866 TI - Validation of the Hopkins Medication Schedule to identify difficulties in taking medications. AB - BACKGROUND: Medications often represent the first line of defense in preventing or delaying the progression of chronic diseases. The health implications of improper medication compliance, and failure to identify it, are considerable. The authors thus developed and validated the Hopkins Medication Schedule (HMS), a new objective test of one's ability to understand and implement a routine prescription medication. METHODS: The authors gave a hypothetical physician's prescription for two common medications (antibiotics and aspirin) to 360 high functioning, community-dwelling, older participants in the Women's Health and Aging Study II and asked them to fill in a daily schedule for taking these medications and to fill in the compartments of a daily pillbox. These scored and timed performances were evaluated for their ability to predict concurrent and 3 year participant-reported difficulty in performing instrumental activities of daily living (IADLs) and for their associations with memory and attention. RESULTS: Although fewer than 2% of participants reported difficulty in taking medications, nearly 22% were completely unable to complete the schedule, fill the pillbox, or both. The 7% of participants who reported difficulty in any IADL also performed poorly on the HMS. In addition, performance on the schedule and the pillbox predicted concurrent difficulty. In adjusted regression analyses, the schedule was most highly associated with memory and the pillbox with learning and executive function. CONCLUSIONS: The HMS had concurrent validity for participant reported IADL difficulty. Furthermore, in community-dwelling older women who largely reported no difficulty in taking medications, the HMS identified nearly 22% who could not write or implement a routine medication regimen. This standardized measure may identify those at increased risk for poor medication adherence and, more broadly, IADL difficulty. PMID- 15814867 TI - Grip strength, postural control, and functional leg power in a representative cohort of British men and women: associations with physical activity, health status, and socioeconomic conditions. AB - BACKGROUND: Understanding the health, behavioral, and social factors that influence physical performance in midlife may provide clues to the origins of frailty in old age and the future health of elderly populations. The authors evaluated muscle strength, postural control, and chair rise performance in a large representative prospective cohort of 53-year-old British men and women in relation to functional limitations, body size, health and activity, and socioeconomic conditions. METHODS: Nurses interviewed 2984 men and women in their own homes in England, Scotland, and Wales and conducted physical examinations in 2956 of them. Objective measures were height, weight, and three physical performance tests: handgrip strength, one-legged standing balance time, and time to complete 10 chair rises. Functional limitations (difficulties walking, stair climbing, gripping, and falls), health status, physical activity, and social class were obtained using a structured questionnaire. RESULTS: Those with the worst scores on the physical performance tests had higher rates of functional limitations for both upper and lower limbs. Women had much weaker handgrip strength, somewhat poorer balance time, and only slightly poorer chair rise time compared with men. In women, health problems and low levels of physical activity contributed to poor physical performance on all three measures. In men, physical activity was the predominant influence. Heavier weight and poorer socioeconomic conditions contributed to poorer balance and chair rise times. CONCLUSIONS: In this representative middle-aged group, physical performance levels varied widely, and women were seriously disadvantaged compared with men. In general, physical performance was worse for men and women living in poorer socioeconomic conditions with greater body weight, poorer health status, and inactive lifestyles. These findings support recommendations for controlling excess body weight, effective health interventions, and the maintenance of active lifestyles during aging. PMID- 15814868 TI - Muscle coordination during rapid force production by young and older adults. AB - BACKGROUND: Older adults typically exhibit dramatic reductions in the rate of force development and deficits in the execution of rapid coordinated movements. The purpose of the current study was to investigate the association between the reduced rate of force development exhibited by older adults and the ability to coordinate groups of muscles. METHODS: The performance of a visually guided aiming task that required the generation of isometric torque about the elbow joint was compared in 10 young adults (age range, 19 to 29 years) and 10 older adults (age range, 65 to 80 years). Participants were required to exert isometric torque in flexion, extension, pronation, or supination, or in combinations of these directions, to reach a target in minimum time. Surface electromyograms were obtained from the biceps brachii, triceps brachii, brachioradialis, and flexor carpi radialis. RESULTS: Older participants exhibited slower target acquisition times compared with young participants (p<.05), with the extent of the differences between the groups varying markedly between target locations. CONCLUSIONS: The impairment in performance, although partially attributable to a general decline in the ability to produce force rapidly, was also affected by the requirements for muscular coordination. At the neuromuscular level, differences between the young and the elderly were expressed most prominently in the bifunctional muscle biceps brachii and in certain temporal aspects of muscular coordination. PMID- 15814869 TI - Prevalence and determinants of impaired glucose metabolism in frail elderly patients: the Belgian Elderly Diabetes Survey (BEDS). AB - BACKGROUND: Although diabetes in elderly persons is generally type 2, the metabolic abnormalities associated with aging suggest that elderly persons may differ from younger persons with type 2 diabetes. In addition, nonobese elderly persons with type 2 diabetes show a marked impairment in insulin release accompanied by mild insulin resistance, whereas obese elderly persons have marked insulin resistance in the presence of "adequate" levels of insulin. Other factors that could adversely affect glucose tolerance in aging include drug use, associated disease, and other stressful conditions commonly encountered in geriatric inpatients units. The authors' objectives in this study were 1) to prospectively assess the prevalence of glucose homeostasis abnormalities among elderly hospitalized patients and the degree to which it reflects abnormalities in insulin secretion or insulin sensitivity using homeostasis model assessment of fasting glucose, insulin, and C-peptide; and 2) to define the social, functional, pathologic, and nutritional characteristics of persons with impaired glucose tolerance or diabetes. METHODS: Ninety-eight patients underwent a comprehensive geriatric assessment. Determinants of glucose homeostasis were assessed using the homeostasis model assessment, which provides estimates of beta-cell function (%B) and insulin sensitivity (%S). RESULTS: Twelve patients (12%) had fasting glucose concentrations greater than 110 mg/dl. Four patients had impaired fasting glucose levels greater than 110 mg/dl but less than 126 mg/dl (IFG group), and 8 patients had levels greater than 126 mg/dl (type 2 diabetes group). Except for a higher proportion of women in the IFG-diabetes group, the latter did not exhibit significant differences in functional, morbidity, or nutritional characteristics compared with the normal glucose tolerance group. The entire cohort (n=98) presented with a mean (+/-SD) %B of 71%+/-47% and a mean %S of 208%+/-198%. Compared with the normal glucose tolerance group, the IFG-diabetes group had a fasting glycemia level of 142+/-24 mg/dl (vs 92+/-9 mg/dl), a %B of 43%+/-21% (vs 74%+/-45%), and a mean %S of 126%+/-113% (vs 219%+/-205%). CONCLUSIONS: These data confirm the high prevalence of impaired glucose metabolism among elderly people, although the usual risk factors were not significantly increased. Marked beta secretory defects seem to be the rule, whereas a significant degree of insulin resistance is unusual. PMID- 15814870 TI - Diseases and impairments as risk factors for onset of disability in the older population in England and Wales: findings from the Medical Research Council Cognitive Function and Ageing Study. AB - BACKGROUND: This paper reports the association between self-reported diseases and impairments and 2-year onset of disability in a prospective study of people aged 65 years or older in five urban and rural centers in England and Wales (Medical Research Council Cognitive Function and Ageing Study; MRC-CFAS). METHODS: We initially reviewed risk factors for onset of disability in 35 prospective studies of functional decline in older people published in 1998-2001. In the present study, disability was defined as requiring help from another person at least several times a week and was assessed by dependency in activities of daily living. Polytomous and bivariate logistic regression models were fitted for onset of disability and mortality among those nondisabled at baseline (n=7913), adjusting for age, sex, and sociodemography. RESULTS: Among prevalent conditions, arthritis (population-attributable risk 11.4%) and cognitive impairment indicated by a Mini-Mental State Examination score of 60 years. The V(MCA)/V(ICA) index was calculated based on angle-corrected blood flow velocities determined in the MCA and extracranial ICA. RESULTS: Mean flow velocities in the MCA and ICA diminished with increasing age, most pronounced in those subjects >40 years. The V(MCA)/V(ICA) index increased significantly (1.67 + 0.005 [age]; P < .05) with age in women, but not in men. In women, reference values and ranges for the V(MCA)/V(ICA) index were as follows, by group: I, 1.82 (range, 0.88-2.68); II, 1.91 (range, 0.94-2.88); and III, 2.06 (range, 0.59-3.53). Respective values for men were as follows, by group: I, 2.10 (range, 0.96-3.24); II, 2.04 (range, 0.71-3.37); and III, 1.78 (range, 0.81-2.75). In subjects <40 years, the V(MCA)/V(ICA) index was significantly higher in men than in women. CONCLUSION: The V(MCA)/V(ICA) index significantly varies with age and sex. Sonographic diagnosis of cerebral vasospasm should be based on age- and sex-adjusted reference values of the V(MCA)/V(ICA) index. PMID- 15814914 TI - Magnetic susceptibility-weighted MR phase imaging of the human brain. AB - BACKGROUND AND PURPOSE: MR gradient echo imaging is sensitive to the magnetic susceptibility of different tissue types. The purpose of this study was to investigate the diagnostic potential of MR phase imaging of the human brain. METHODS: High-spatial-resolution, T2*-weighted, single-echo images were acquired in five volunteers and one patient with a brain tumor on a 1.5T system by applying a 3D, first-order, velocity-compensated gradient echo sequence by using a quadrature transmit-receive head coil. Phase images were reconstructed from the raw data and unwrapped by using a region-growing phase-unwrapping algorithm. Low spatial-frequency components originating from static background susceptibility effects were removed by high-pass filtering. RESULTS: Phase images showed excellent image contrast and revealed anatomic structures that were not visible on the corresponding magnitude images. CONCLUSION: Improved processing of susceptibility-weighted MR phase images offers a new means of contrast for neuroimaging applications. PMID- 15814916 TI - Persistent falcine sinus in an adult: demonstration by MR venography. AB - The falcine sinus is a normal anatomic structure located in the falx cerebri that closes after birth and is rarely observed in the adult population. We describe a unique case of persistent falcine sinus in an adult who presented with venous sinus obstruction. PMID- 15814915 TI - PC VIPR: a high-speed 3D phase-contrast method for flow quantification and high resolution angiography. AB - BACKGROUND AND PURPOSE: Three-dimensional phase-contrast (3DPC) is limited by long imaging times, limited coverage, flow artifacts, and the need to perform multiple additional 2D examinations (2DPC) to measure flow. A highly undersampled 3D radial acquisition (isotropic-voxel radial projection imaging [PCVIPR]) makes it possible to increase the product of volume coverage and spatial resolution by a factor of 30 for the same imaging time as conventional Cartesian 3DPC. This provides anatomic information over a large volume with high isotropic resolution and permits retrospective measurement of average flow rates throughout the volume. METHODS: PCVIPR acquires a reference and three flow-encoded acquisitions for each VIPR projection. Complex difference images were formed by combining information from all flow directions. Following retrospective definition of planes perpendicular to selected vessels, volume flow rates were determined by using phase-difference information. The accuracy of average flow measurement was investigated in a phantom and in six volunteers. Anatomic PCVIPR images acquired in three patients and three volunteers by using a 384(3) matrix were compared with conventional Cartesian 3DPC. RESULTS: The flow validation produced R2 = 0.99 in vitro and R2 = 0.97 in vivo. PCVIPR produced minimal streak and pulsatile flow artifacts. PCVIPR produced far higher resolution and volume coverage in comparable imaging times. The highest acceleration factors relative to 3DPC were achieved by using gadolinium-contrast material. Ultimately, acceleration factors are limited by signal-to-noise ratio. CONCLUSION: PCVIPR rapidly provides isotropic high-resolution angiographic images and permits retrospective measurement of average flow rate throughout the volume without the need to prescribe multiple 2D acquisition planes. PMID- 15814918 TI - Relationship of MR-derived lactate, mobile lipids, and relative blood volume for gliomas in vivo. AB - BACKGROUND AND PURPOSE: Gliomas are heterogeneous tumors with increased microvasculature, hypoxia, and necrosis. The purpose of this study was to examine the distribution and relationship of the MR-derived relative cerebral blood volume (rCBV), lactate (Lac), and mobile lipids (Lip), which have been proposed as markers for these phenomena of gliomas in vivo. METHODS: Twenty-three patients with newly diagnosed gliomas were examined before surgical biopsy and/or resection (seven grade II, five grade III, and 11 grade IV), and 27 patients were studied after surgery but before radiation treatment and/or chemotherapy (11 grade II, two grade III, and 14 grade IV gliomas). Lac and Lip were estimated from Lac-edited three-dimensional MR spectroscopic images. Dynamic susceptibility contrast MR imaging was applied to obtain perfusion-weighted images and rCBV maps. RESULTS: Before surgical biopsy/resection, one low-grade and 12 of 16 high grade gliomas had significantly elevated Lac. No low-grade and 10 of 16 high grade gliomas had significantly elevated Lip. In presurgical high-grade gliomas, volumes of elevated Lip and macronecrosis were significantly correlated. rCBV was significantly increased in regions with elevated Lac. After we excluded macronecrosis, the rCBV for regions with elevated Lip but no Lac was significantly higher than rCBV in regions with elevated Lac but no Lip. After surgical biopsy/resection, more low-grade patients showed significantly elevated Lac and Lip than before. CONCLUSION: The in vivo distribution of rCBV, Lac, and Lip, as evaluated with three-dimensional MR spectroscopic imaging, may help in the diagnosis and selection of the most appropriate therapy for patients with gliomas. PMID- 15814917 TI - Relationships between astrogliosis and 1H MR spectroscopic measures of brain choline/creatine and myo-inositol/creatine in a primate model. AB - BACKGROUND AND PURPOSE: In vivo 1H MR spectroscopy demonstrates elevated choline (Cho)/creatine (Cr) and myo-inositol (MI)/Cr in many neurologic diseases that has been ascribed to gliosis. We tested the hypotheses that in vivo Cho/Cr and/or MI/Cr levels are correlated with glial fibrillary acidic protein (GFAP) immunostains and that the changes are water-soluble metabolites. METHODS: We performed postmortem 1H MR spectroscopy and GFAP immunohistochemistry in brains from seven rhesus macaques acutely infected with simian immunodeficiency virus (SIV) and in four controls and compared the findings with previous in vivo MR spectroscopic results. Changes in neuropathologic and MR spectroscopic markers after infection and relationships among plasma viral load, GFAP immunostaining results, and ex vivo and in vivo MR spectroscopic measures were statistically evaluated. RESULTS: On GFAP immunostaining and in vivo MR spectroscopy, GFAP, Cho/Cr and MI/Cr were highest near the time of peak plasma viral load at 11 days postinfection (dpi). Immunostains returned to baseline by 14 dpi, whereas Cho/Cr and MI/Cr had different time courses, with the former dropping below baseline and the latter remaining elevated. Viral load and immunostains were significantly correlated. No correlation was found between ex vivo Cho/Cr or MI/Cr and viral load or between metabolite ratios from in vivo and ex vivo MR spectroscopy. CONCLUSION: In acute SIV infection, plasma viral load was significantly correlated with brain GFAP immunostains and in vivo 1H MR spectroscopic Cho/Cr. In vivo changes in Cho/Cr and MI/Cr were principally due to contributions other than those of low-molecular-weight water-soluble metabolites. PMID- 15814919 TI - Measurement of tumor "size" in recurrent malignant glioma: 1D, 2D, or 3D? AB - BACKGROUND AND PURPOSE: Tumor "size" is used internationally as a surrogate marker for overall survival when following current response assessment protocols (World Health Organization and Response Evaluation Criteria in Solid Tumors). With little evidence of a relationship between tumor "size" and survival in intrinsic brain tumors, this study was undertaken to investigate the predictive value of MR imaging-defined tumor size for survival in patients with recurrent malignant glioma and to compare the different measures of tumor size used in these current response assessment protocols. METHODS: Volumetric, bidimensional, and unidimensional measurements of tumor size were made using baseline contrast enhanced T1-weighted images of 70 patients with recurrent malignant glioma receiving intravenous chemotherapy. Cox's proportional hazards model was used to investigate the prognostic importance of tumor size using survival as the end point. Further statistical analysis was undertaken to investigate the relationship between the different measurement techniques. RESULTS: Only the volumetric measurement of tumor size was found to be predictive of survival in recurrent malignant glioma on both univariate and multivariate analysis. Furthermore, analysis demonstrated that the unidimensional and bidimensional measures of tumor were not comparable with the more accurate and direct volumetric measurement. CONCLUSION: Indirect unidimensional and bidimensional measurement techniques do not have a significant association with overall survival or adequately assess tumor size in recurrent malignant glioma. These findings have serious implications about the validity of using current response assessment protocols in therapy trials for recurrent malignant glioma. PMID- 15814920 TI - MR cerebral blood volume maps correlated with vascular endothelial growth factor expression and tumor grade in nonenhancing gliomas. AB - BACKGROUND AND PURPOSE: Relative cerebral blood volume (rCBV) measurements derived from perfusion-weighted imaging (PWI) may be useful to evaluate angiogenesis and preoperatively estimate the grade of a glioma. We hypothesized that rCBV is correlated with vascular endothelial growth factor (VEGF) expression as marker of the angiogenic stimulus in presumed supratentorial low-grade gliomas (LGGs). METHODS: From February 2001 to February 2004, we examined 20 adults (16 men, four women; mean age 36 years; range, 23-60 years) with suspected (nonenhancing) supratentorial LGG on conventional MR imaging. Preoperative MR imaging used a dynamic first-pass gadolinium-enhanced, spin-echo echo-planar PWI. In heterogeneous tumors, we performed stereotactic biopsy in the high-perfusion areas before surgical resection. Semiquantitative grading of VEGF immunoreactivity was applied. RESULTS: Nine patients had diffuse astrocytomas (World Health Organization grade II), and 11 had other LGG and anaplastic gliomas. In patients with heterogeneous tumors on PWI, the high-rCBV focus had areas of oligodendroglioma or anaplastic astrocytoma on stereotactic biopsy, whereas the surgical specimens were predominantly astrocytomas. Anaplastic gliomas had high rCBV ratios and positive VEGF immunoreactivity. Diffuse astrocytomas had negative VEGF expression and mean rCBV values significantly lower than those of the other two groups. Three diffuse astrocytomas had positive VEGF immunoreactivity and high rCBV values. CONCLUSION: Our results confirmed the correlation among rCBV measurements, VEGF expression, and histopathologic grade in nonenhancing gliomas. PWI may add useful data to the preoperative assessment of nonenhancing gliomas. Its contribution in predicting tumor behavior and patient prognosis remains to be determined. PMID- 15814921 TI - Can tumor contrast enhancement be used as a criterion for differentiating tumor grades of oligodendrogliomas? AB - BACKGROUND AND PURPOSE: The association of high-grade oligodendrogliomas with tumor contrast material enhancement on MR images has been reported. Some authors have even used contrast enhancement as a criterion for their oligodendroglioma grading system. The purpose of our study was to evaluate if tumor contrast enhancement is a specific finding for anaplastic oligodendroglioma. METHODS: Pretreatment MR images of 24 oligodendrogliomas were reviewed retrospectively, and findings were compared with the histologic grade. The presence or absence and the pattern of tumor contrast enhancement were evaluated qualitatively. A contrast enhancement ratio (CER), a quantitative criterion, was calculated to assess the difference in degree of enhancement between the low-grade and anaplastic tumors. Tumor grade was diagnosed at pathologic examination according to the World Health Organization classification system. RESULTS: Contrast enhancement was noted in nine (56%) of 16 low-grade tumors and in five (62%) of eight anaplastic tumors. A characteristic enhancement pattern, nodular-like enhancement, was found in eight tumors. The CERs were 2.12-40.88 (mean, 20.08) in low-grade tumors and were 3.20-62.52 (mean, 28.73) in anaplastic tumors (P > .05). CONCLUSION: Tumor contrast enhancement was not statistically significantly different between the tumor groups. We believe that the presence or absence of tumor contrast enhancement is not a specific finding for simply discriminating low-grade from anaplastic oligodendrogliomas. Histologic confirmation is necessary even in tumors without contrast enhancement. PMID- 15814922 TI - Diffusion tensor imaging of the corticospinal tract before and after mass resection as correlated with clinical motor findings: preliminary data. AB - BACKGROUND AND PURPOSE: The role of diffusion tensor imaging (DTI) in neurosurgical planning and follow-up is currently being defined and needs clinical validation. To that end, we sought correlations between preoperative and postoperative DTI and clinical motor deficits in patients with space-occupying lesions involving the corticospinal tract (CST). METHODS: DTI findings in four patients with masses near the CST and not involving motor cortex were retrospectively reviewed and compared with contralateral motor strength. CST involvement was determined from anisotropy and eigenvector directional color maps. The CST was considered involved if it was substantially deviated or had decreased anisotropy. Interpretations of the DTIs were blinded to assessments of motor strength, and vice versa. RESULTS: Of the four patients with potential CST involvement before surgery, DTI confirmed CST involvement in three, all of whom had preoperative motor deficits. The patient without CST involvement on DTI had no motor deficit. After surgery, DTI showed CST preservation and normalization of the position and/or anisotropy in two of the three patients with preoperative deficits, and both of those patients had improvement in motor strength. The other patient with preoperative deficits had evidence of wallerian degeneration on DTI and had only equivocal clinical improvement. CONCLUSION: Preoperative CST involvement, as determined on DTI, was predictive of the presence or absence of motor deficits, and postoperative CST normalization on DTI was predictive of clinical improvement. Further study is warranted to define the role of DTI in planning tumor resections and predicting postoperative motor function. PMID- 15814923 TI - Diffusion tensor imaging in cases with visual field defect after anterior temporal lobectomy. AB - BACKGROUND AND PURPOSE: Visual defect due to optic radiation injury is a complication of temporal lobectomy for temporal epilepsy. To investigate whether diffusion tensor imaging can delineate the changes in optic radiations after lobectomy, we evaluated parameters on tensor images for optic radiations and correlated them with visual defect. METHODS: We examined 14 cases after temporal lobectomy. Durations after surgeries ranged from 3 weeks to 9 years. The cases were classified into three groups on the basis of the severity of visual field defect (A-C, with group C the most severe). We evaluated signals on T2-weighted images and parameters of tensor images, including fractional anisotropy (FA) and apparent diffusion coefficient (ADC), for the optic radiation in both the operated and intact side. RESULTS: On T2-weighted images, high signals in optic radiations were seen in four cases, occurring more than 4 weeks after surgery. The mean operated-to-intact side FA ratio in the optic radiation decreased according to severity of visual defect (group A, 0.88; group B, 0.89; group C, 0.73). The mean operated-to-intact side ADC ratio showed no significant difference in the overall cases. The ratio for ADC, however, tended to increase according to visual defect in cases after 10 weeks postsurgery. CONCLUSION: Optic radiation showed a decreased FA value in cases after temporal lobectomy. In later stages, ADC values tended to increase and high signal intensities on T2-weighted images were observed. The FA value can be used for evaluating Wallerian degeneration of optic radiation even in the early stages after surgery. PMID- 15814924 TI - MR Imaging of pial melanosis secondary to a posterior fossa melanotic ependymoma. AB - A 36-year-old man presented with trouble speaking and bilateral progressive hearing loss. MR imaging and histopathologic results revealed a posterior fossa melanotic ependymoma. Pial surfaces appeared hyperintense on T1-weighted images and hypointense on T2-weighted images. Histopathologic examination revealed that tumor cells and interstitial spaces had abundant melanin accumulation. There was no evidence of hemosiderin in tumor cells and in interstitial spaces. Pial melanin accumulation secondary to a posterior fossa melanotic ependymoma explained our MR findings. PMID- 15814925 TI - Mechanisms of bihemispheric brain infarctions in the anterior circulation on diffusion-weighted images. AB - BACKGROUND AND PURPOSE: Multiple acute brain infarctions in both cerebral hemispheres usually suggest an embolic mechanism, particularly one of aortic or cardiac origin. The purpose of this study was to clarify the etiologic mechanisms and topographic features of bihemispheric infarctions depicted on diffusion weighted imaging (DWI). METHODS: Among 411 consecutive patients with ischemic stroke who underwent MR imaging in the acute phase, DWI showed bilateral infarctions in 19 (4.6%). In these patients, we analyzed the presence of carotid, aortic or cardiac embolic sources by using ultrasonography, cerebral angiography, and/or transesophageal echocardiography and evaluated the size and topographic distribution of the lesions. We assessed intracranial cross-flow through the anterior communicating artery, mainly on the basis of the anatomic information obtained from angiography or MR angiography. RESULTS: Bilateral lesions were derived from cardiac and/or aortic embolic sources in 16 (84%) of 19 patients and appeared to originate from unilateral carotid diseases in three (16%). In nine (82%) of 11 patients with cardiac embolic sources, at least one large territorial or subcortical lesion was found in either hemisphere, whereas in all eight patients without a cardiac embolic source, the lesions were small and disseminated bilaterally. CONCLUSION: Unilateral carotid lesions can cause bihemispheric infarctions through cross-flow in the anterior communicating artery. On DWI, small bihemispheric, disseminated lesions strongly suggest an artery-to-artery embolism. In such cases, aortic and carotid lesions should be assessed as potential embolic sources. PMID- 15814926 TI - Tissue at risk is overestimated in perfusion-weighted imaging: MR imaging in acute stroke patients without vessel recanalization. AB - BACKGROUND AND PURPOSE: The volume of decreased cerebral blood flow (CBF) in acute stroke perfusion-weighted imaging frequently overestimates final infarct volume. We hypothesized that surviving tissue exists even in patients without recanalization and tried to determine perfusion thresholds from initial MR imaging. METHODS: Stroke MR imaging including MR angiography was carried out at days 0, 1, and 7 after stroke onset in 19 patients without recanalization at least until day 1. The following lesions were defined: L0 = diffusion restriction at day 0; LG1 = lesion growth until day 1; LG7 = lesion growth until day 7; ST7 = initially hypoperfused, but surviving tissue. These lesions were transferred on initial MR imaging within 4.7 hours and perfusion values at day 0 were determined. RESULTS: Median lesion volume L0 at day 0 was 18.2 mL and increased to 39.4 and 43.8 mL at days 1 and 7. Volume of decreased rCBF not progressing to infarction was 148.5 mL (ST7). Mean ST7 perfusion values were different from L0 and LG1, but only mean relative cerebral blood volume (rCBV) was different from LG7, discriminating survival against death of tissue. A threshold value of 0.82 CBV for death versus survival was determined with a sensitivity of 0.56 and specificity of 0.95. Carotid T occlusions showed the greatest potential of lesion growth. CONCLUSION: Even when vessel occlusion persists, hypoperfused tissue on MR imaging does not necessarily progress toward infarction. The most conclusive inferences can be drawn from CBV images. The site of arterial occlusion also determines progression to infarction. PMID- 15814927 TI - Clearing of red blood cells in lumbar puncture does not rule out ruptured aneurysm in patients with suspected subarachnoid hemorrhage but negative head CT findings. AB - BACKGROUND AND PURPOSE: In evaluating the results of lumbar puncture (LP), a decrease in the number of red blood cells (RBCs)/mm3 between the first and fourth tubes collected (clearing) has often been assumed to indicate a traumatic puncture rather than the presence of subarachnoid hemorrhage (SAH). We tested the hypothesis that, in the setting of severe headache, CSF clearing coupled with an unremarkable unenhanced CT scan was negatively predictive of the presence of aneurysm and could be used to reduce the need for conventional arteriography. METHODS: Cerebral angiography was performed to evaluate suspected SAH in 123 consecutive patients over 2 years at a university teaching hospital. Records of these patients were reviewed. Among the subset without SAH on CT scan, LP results were evaluated for clearing. Clearing was arbitrarily defined as a 25% reduction in RBCs between the first and fourth tubes. This subset's records were also reviewed for the presence of aneurysm at cerebral angiography or at follow-up 6 weeks later. Data were analyzed for correlation between clearing and aneurysm. RESULTS: Of the 123 patients whose records were reviewed, 22 did not show an SAH on CT scan. Of those 22 patients, eight had aneurysm at angiography and 14 did not. Clinical diagnoses in the other 14 included trauma, herpes meningitis, sickle cell disease, and cocaine use. CSF clearing was noted in 25% of those with an aneurysm (two of 8) and 21% of those without an aneurysm (three of 14). In the two cases with aneurysms, RBCs cleared from 3550 to 2550 (-28%) and from 24,686 to 17,842 (-28%), respectively. In the remaining six cases with aneurysms, RBCs increased a mean of 1370% between the first and fourth tubes (range, -22% to 7700%). Two of these six had a reduction that did not meet our criteria for clearing (-22% and -5.3%, respectively). In the 14 cases without aneurysms, RBCs increased a mean of 70% between the first and fourth tubes (range, -99% to 895%). In addition to the three of these 14 that met our criteria for clearing (-99%, 99%, and -43%), four cases had a reduction that did not (range, -0.7% to -14%). CONCLUSION: A 25% reduction in RBC concentration between the first and fourth tubes of CSF in patients with suspected SAH but negative CT findings occurs even in cases of ruptured aneurysms. Formal evaluation for the presence of an aneurysm is still necessary in this scenario. PMID- 15814928 TI - Hemodynamic and permeability changes in posterior reversible encephalopathy syndrome measured by dynamic susceptibility perfusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: The physiological mechanism that gives rise to posterior reversible encephalopathy syndrome (PRES) is currently unknown. We sought to better understand the mechanism of the edema formation in PRES and specifically hypothesized that this edema is caused by increased vascular permeability. METHODS: Eight consecutive patients with PRES who had been studied by using perfusion MR imaging were retrospectively identified. Perfusion images were obtained using a gradient-echo echo planar sequence with contrast enhancement. Measures of apparent diffusion coefficient (ADC), cerebral blood volume (CBV), cerebral blood flow (CBF), mean transit time (MTT) and vascular permeability (K2) were calculated in the affected posterior brain areas and normalized to values obtained in unaffected anterior brain. These values were compared with those found in healthy subjects. RESULTS: Regions of interest within the posterior affected brain indicated a significant increase in ADC values in all but one patient, a significant decrease in CBV and CBF values in all patients with PRES (average 30% of control), variable changes in MTT, and no measurable change in K2. CONCLUSION: The decrease in both CBV and CBF in PRES supports the theory of autoregulatory vasoconstriction; changes in K2 were not detected. PMID- 15814929 TI - Persistence of congenital mirror movements after hemiplegic stroke. AB - We describe a case of hereditary congenital mirror movements (MMs) in a 76-year old man, who after an ischemic stroke, had persistence of MMs in the paretic hand during voluntary movements of the contralateral arm. By using functional MR imaging to investigate the performance of motor and sensory tasks with the affected and the unaffected hands, we found evidence for increased ipsilateral primary motor cortex activity and reduced transcallosal inhibition. Both these mechanisms are likely to be involved in the genesis of MMs. PMID- 15814930 TI - MR features of cerebral aspergillosis in an immunocompetent patient: correlation with histology and elemental analysis. AB - We report an unusual case of cerebral aspergillosis in a young immunocompetent patient who also had dissemination to other end organs. The patient presented with a large mass in the left cerebral hemisphere. Elemental analysis of biopsy specimens revealed elevated levels of iron, magnesium, zinc, calcium, chromium, and nickel that correlated with a peripheral rim of hypointensity on T2-weighted images. PMID- 15814931 TI - Brain abscess drainage by use of MR fluoroscopic guidance. AB - We describe herein the use of MR fluoroscopic guidance in the drainage of abscess cavities. We percutaneously drained 12 brain abscesses in 11 patients. A 0.3T open MR imaging system was used. Sixteen drainages were performed in 12 abscesses. Repeat drainage was needed in three recurrences and one residual lesion. No serious complications were seen. MR fluoroscopy-guided percutaneous brain abscess drainage in an open MR imaging system is feasible. PMID- 15814932 TI - Detectability of hypometabolic regions in mild Alzheimer disease: function of time after the injection of 2-[fluorine 18]-fluoro-2-deoxy-D-glucose. AB - BACKGROUND AND PURPOSE: 2-[Fluorine 18]-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) has played an important role in detecting hypometabolic regions in the brains of patients with dementia. To our knowledge, the optimal imaging time for dementia has not been investigated. The aim of this study was to evaluate the sensitivity of the early scanning (ES) compared with late scanning (LS) for demonstrating decreased regional glucose metabolism in patients with Alzheimer disease (AD). METHODS: Twenty patients with mild AD (mean age +/- standard deviation, 64.8 +/- 5.2 years) and 20 age- and sex-matched healthy volunteers (age, 65.9 +/- 4.5 years) were underwent FDG PET. Their cerebral glucose metabolic images were obtained on ES at 30-42 minutes and LS at 60-72 minutes after the administration of FDG 185-346 MBq. We compared regional cerebral metabolic images in a voxel-by-voxel analysis with statistical parametric mapping between patients with AD and control subjects and evaluated the difference in the hypometabolic regions between the two scans. RESULTS: In the AD-to-healthy comparison, LS at the P < .001 level of significance showed more extensive and significant hypometabolic areas than did ES. CONCLUSION: These results indicate that LS is superior to ES in detecting hypometabolic regions in patients with AD. For patients with AD, emission scanning soon after the administration of FDG is probably not advised. PMID- 15814933 TI - MR angiographic evaluation of platinum coil packs at 1.5T and 3T: an in vitro assessment of artifact production: technical note. AB - Susceptibility artifact from platinum coil packs impairs the visibility of perianeurysmal soft tissues at conventional 3D time-of-flight MR angiography. These artifacts limit the evaluation for residual-recurrent aneurysm and parent vessel stenosis. Reducing the echo time can decrease the artifact and improve perianeurysmal visualization. The purpose of this study was to assess quantitatively the effect of decreasing the echo time on artifact production at different field strengths and coil pack densities. PMID- 15814934 TI - Carotid artery stent placement with distal balloon protection: technical considerations. AB - BACKGROUND AND PURPOSE: Most neurologic events that complicate the carotid artery stent placement procedure are embolic. Strategies are being developed and evaluated for their ability to minimize the clinical embolic risk. The purpose of this study was to determine the feasibility, safety, and technical considerations of performing carotid artery stent placement with use of a distal balloon antiembolization system. METHODS: Carotid artery stent placement with use of a commercially available distal balloon antiembolization system was performed in 303 patients (325 vessels). Clinical neurologic complications were adjudicated at the time of intervention and at 1 month. RESULTS: Technical success was achieved in all but one case, with reduction of the mean percentage carotid artery stenosis from 79 +/- 9% to 3 +/- 8%. Intolerance to internal carotid artery occlusion occurred in 11 patients (3.6%); in one patient the procedure was aborted, and in 10 the procedures were expeditiously completed with no adverse neurologic events. The 30-day combined rate of all occurrences of stroke and death was 2%. These events included three (0.9%) retinal emboli, three (0.9%) minor strokes, and one (0.3%) fatal stroke from brain hemorrhage, all of which occurred during the procedure with no events occurring after discharge. In addition, two (0.6%) transient ischemic attacks and three (0.9%) reperfusion syndromes occurred, with the patients experiencing complete recovery. CONCLUSION: Carotid artery stent placement with use of the distal balloon protection system is feasible and safe, and the short-term outcomes appear to be favorable. Optimal application of proper technique is important. PMID- 15814935 TI - A novel flexible, retrievable endovascular stent system for small-vessel anatomy: preliminary in vivo data. AB - BACKGROUND AND PURPOSE: This study assessed the in vivo delivery, retrievability, short-term patency, and cellular response to a new flexible endovascular stent system in a rabbit model. The stent is designed for delivery through a microcatheter and is fully retrievable with electrolytic detachment from a delivery wire. METHODS: We successfully deployed nine stents (range of sizes, 2.5 4 mm diameter, 15-35 mm length) in six straight (carotid) and three angled (subclavian) arteries of six Chinchilla Bastard rabbits. Serial imaging was performed by using intravenous digital subtraction angiography (IVDSA), contrast enhanced MR angiography (CEMRA), time-of-flight MR angiography (TOF), and CT angiography 3 days and 4 weeks after stent deployment. Subjects were euthenized after 4 weeks (n = 5), and stents were removed for histologic analysis. RESULTS: Stent deployment was feasible in all cases. After initial deployment, all stents could be fully retrieved within the microcatheter. The detachment zone and the distal stent marker were easily visible under fluoroscopy, and final detachment occurred reliably in all cases. We observed no procedural complications. Noninvasive imaging by using IVDSA, MR angiography, and CT angiography was feasible in this stent system and demonstrated all arteries patent and not narrowed at 3 days and 4 weeks, findings that were confirmed by histologic analysis. CONCLUSION: This electrolytically detachable stent is promising as a treatment for intracranial arteries, because it can be delivered through microcatheters small enough for intracranial navigation. It is fully retrievable, thus providing greater control than currently available stents. Noninvasive imaging by using IVDSA, MR angiography, and CT angiography is feasible in this stent system and may be useful for follow-up. Further long-term data are needed. PMID- 15814936 TI - Stent-assisted angioplasty of intracranial vertebrobasilar atherosclerosis: midterm analysis of clinical and radiologic predictors of neurological morbidity and mortality. AB - BACKGROUND AND PURPOSE: Initial reports of stent-assisted angioplasty for intracranial vertebrobasilar atherosclerosis suggest this is a feasible treatment, but there have been little data regarding predictors of success or failure. We analyzed a series of patients for independent predictors of neurologic morbidity and mortality. METHODS: Patient charts and angiograms from 39 patients who underwent intracranial angioplasty and stent placement of vertebrobasilar stenoses were retrospectively reviewed to obtain clinical and detailed angiographic data on potential predictors of neurologic morbidity and mortality. Univariate analyses of these predictors were performed with either Fisher's exact test or simple logistic regression. Multivariate analysis was subsequently performed on the statistically significant predictors. RESULTS: Complete clinical data were obtained for 39 patients, and angiographic review was possible for 35 of them. Angiography revealed severe intracranial vertebral (n = 18), basilar (n = 15), or basilar and vertebral (n = 2) stenoses. Two patients (5.1%) died in the periprocedural period, nine patients (23.1%) had neurologic complications, and one patient (2.6%) had transient neurologic symptoms. Univariate analysis revealed female sex, diabetes, and failure of coumadin or heparin therapy were associated with neurologic morbidity, whereas female sex, Mori B lesion, and length-to-stenosis ratio were associated with mortality. The presence of diabetes was the only independent predictor of neurologic morbidity and mortality. CONCLUSION: Because of the limited number of patients available for analysis, the only independent predictor of neurologic morbidity and mortality was diabetes, but several other predictors showed trends that deserve further review in future series. PMID- 15814937 TI - Mechanical thrombectomy for acute stroke. AB - BACKGROUND AND PURPOSE: We evaluated a mechanical thrombectomy protocol to treat acute stroke and report the angiographic results and clinical outcomes. METHODS: Patients with anterior circulation strokes <8 hours and posterior circulation strokes <12 hours were treated at a single center over 10 months. Patients were excluded if they were candidates for intravenous tissue plasminogen activator (tPA). Treatment involved one of two mechanical thrombectomy devices. Retrieval was augmented by low-dose intra-arterial tPA if needed. Outcome was measured by using the Modified Rankin score. RESULTS: Ten patients were treated: five with anterior circulation strokes, four with posterior circulation strokes, and one with embolic strokes involving both circulations. Mean National Institutes of Health Stroke Scale score at presentation was 24.6 +/- 10.9. In eight patients (80%), revascularization was successful (Thrombolysis in Acute Myocardial Infarction score, 3). Mean time from symptom onset to initiation of the procedure was 6 hours (5.3 hours for anterior circulation and 7.0 hours for posterior circulation). Mean time for recanalization from the start of the procedure was 1.17 +/- 0.58 hours for the six anterior circulation strokes and 2.75 +/- 1.34 hours in the two posterior circulation strokes. Five patients died within 48 hours; all had posterior circulation strokes. Mean Modified Rankin score at 90 days was 1.4. CONCLUSION: In this small series, mechanical thrombectomy of acute stroke appeared to improve recanalization rates compared with intra-arterial thrombolysis. No hemorrhagic complications occurred. Further study is required to determine the role of these techniques. PMID- 15814938 TI - Thrombolytic therapy of acute ischemic stroke: correlation of angiographic recanalization with clinical outcome. AB - BACKGROUND AND PURPOSE: The effect of vessel patency, following recombinant tissue plasminogen activator (rtPA) administration, on clinical outcome in acute ischemic stroke (AIS) has been controversial. We studied the effect of recanalization following intraarterial (IA) and intravenous/IA (IV/IA) rtPA on clinical outcome in AIS. METHODS: Recanalization was classified angiographically as complete (as compared with unoccluded vessel, thrombolysis in myocardial infarction classification [TIMI] 3), none (with no change from prethrombolysis, TIMI 0), and partial (when a change in the flow from baseline was noted, TIMI 1 2). Outcomes were symptomatic intracranial hemorrhage (sICH), 90-day modified Rankin scale (< or = 2 as a good outcome), and 3-month mortality. RESULTS: Ninety six patients had either combined IV/IA (41) or IA (55) rtPA for AIS during a 7 year period. Any recanalization occurred in 69%; 55% of those had a good outcome versus 23% in the rest (Odds ratio = 3.9; 95% confidence interval [CI] = 1.4 11.2; P = .007). Only 24% had complete recanalization; 74% had a good outcome versus 36% in the nonrecanalization group (OR = 5.1; 95% CI = 1.6-16.8; P = .002). When adjusted to time to therapy and vessel occluded, these results lessened but remained significant. The sICH rate with any recanalization was 7.6% versus 13.3% in patients with persistent clot (relative risk (RR) = 0.6; 95% CI = 0.2-2.0; P = .45). Death occurred in 19.7% of those whose vessels recanalized versus 33.3% in the rest (RR = 0.56; 95% = 0.26-1.19; P = .2). CONCLUSION: A total of 24% and 69% of patients had complete and any recanalization, respectively, following endovascular rtPA therapy of AIS. The degree of recanalization was directly related to time to therapy and associated with good clinical outcome without an increase in the rate of adverse effect. PMID- 15814939 TI - Selective endovascular treatment of intracranial aneurysms with a liquid embolic: a single-center experience in 39 patients with 41 aneurysms. AB - BACKGROUND AND PURPOSE: Although embolization with detachable coils is an accepted alternative to surgical clipping, a major long-term problem is aneurysm recanalization due to coil compaction. Liquid embolic agents are a possible alternative as filling material that might decrease the recanalization rate. We evaluated the use of a liquid embolic for endovascular treatment of intracranial aneurysms. METHODS: During 1999-2003, 10 patients with 11 small aneurysms (group 1) and 29 patients with 30 large or giant aneurysms (group 2) were treated with a liquid embolic. Of 32 female and seven male patients, 20 had mass effect and two had subarachnoid hemorrhage; 17 were asymptomatic. All aneurysms were judged unsuitable for regular treatment; selective embolization was performed with a liquid embolic alone or with coils and liquid embolic. Stent placement was performed in 15 cases. Clinical and anatomic outcomes were assessed with the Modified Glasgow Outcome Scale and with angiography at 3, 12, and 24 months. RESULTS: In group 1, good or excellent outcome and complete occlusion were observed in all patients. In group 2, clinical outcome was good or excellent in 26 patients and fair in one, and death occurred in two patients (one procedure related and one disease related). Technical complications occurred in four patients in group 1 (one permanent neurologic deficit) and in four patients in group 2 (one patient died, two remain hemiparetic, one remains asymptomatic). Follow-up images showed two recanalizations in group 1 and nine in group 2. CONCLUSION: Selective embolization with a liquid embolic is useful to treat aneurysms unsuitable for coiling or for patients in whom previous treatment failed. This mostly applies to large and giant aneurysms in which morbidity and mortality rates are better than those associated with surgery, and the recanalization rate is lower than that previously described with coiling. PMID- 15814940 TI - Endovascular treatment of wide-necked aneurysms by using two microcatheters: techniques and outcomes in 25 patients. AB - BACKGROUND AND PURPOSE: The endovascular occlusion of aneurysms with unfavorable configurations such as a broad neck and an important branch from the fundus remains a technical challenge. The purpose of this study was to evaluate the radiologic and clinical results of complicated aneurysm treatment by using two microcatheters. METHODS: Twenty-five aneurysms in 25 patients were treated by using two microcatheters, from August 2001 to February 2004. Fourteen patients presented with a subarachnoid hemorrhage (SAH) and 11 had unruptured aneurysms. The aneurysms were of the basilar top (7), middle cerebral artery bifurcation (4), posterior communicating artery (4), anterior communicating artery (3), superior cerebellar artery (2), ophthalmic artery (2), and one aneurysm of each of cavernous internal carotid artery (ICA), dorsal ICA, and midbasilar artery. In 16 aneurysms (64%), the width of the aneurysm was the same or longer than the height. In 19 (76%), important branches arose from the aneurysm base, and some were even incorporated with the aneurysm fundus. The mean dome (height)-to-neck ratio was 1.23 +/- 0.37 (range, 0.65-2.33), and this was greater than or equal to 1.0 in 19 aneurysms (76%). RESULTS: All aneurysms were successfully embolized. Immediate postembolization angiography showed no residual contrast filling in eight aneurysms (32%), and some residual contrast filling in 16. The aneurysm remnants, however, were intentionally left to preserve important branches in 12 of the 16 aneurysms with incomplete occlusion. Two complications occurred, including one thromboembolic and one coil protrusion, but they were successfully resolved and produced no clinical symptoms. All patients except one showed excellent clinical outcomes. One patient revealed moderate cognitive dysfunction. During the follow-up period, no new bleeding occurred. CONCLUSION: Our experience with 25 cerebral aneurysm patients shows that the technique of using two microcatheters is feasible and safe for coil embolization of aneurysms with unfavorable configurations. Although the lack of angiographic follow-up prevents us from drawing conclusions about its effectiveness as compared with other techniques such as stent placement and balloon-neck protection, we believe that this technique offers a reliable alternative for endovascular therapy of complicated aneurysms. PMID- 15814941 TI - Coil thickness and packing of cerebral aneurysms: a comparative study of two types of coils. AB - BACKGROUND AND PURPOSE: In coiling cerebral aneurysms, high packing prevents reopening over time. The purpose of this study was to compare packing of cerebral aneurysms treated with two types of coils with different wire thickness and different shapes. METHODS: Packing, defined as the ratio between the volume of inserted coils and the volume of an aneurysm, was calculated for 144 cerebral aneurysms treated in 130 patients. Seventy-two aneurysms were treated with predominantly helical-shaped coils of 0.010-inch-diameter wire, and 72 aneurysms were treated with predominantly complex-shaped coils of 0.012-inch-diameter wire. Aneurysm volume was assessed from three-dimensional angiography. Aneurysm packing, inserted coil lengths, and numbers of coils were compared for both types of coils. RESULTS: Mean packing was significantly higher (absolute value, 6.4%; relative value, 26.6%; P < .0001) in aneurysms coiled with 0.012-inch-diameter coils than aneurysms coiled with 0.010-inch-diameter coils. Inserted coil length per cubic millimeter of aneurysmal volume was equal for both types of coils. CONCLUSION: Use of complex-shaped coils with a wire diameter of 0.012 inch to coil cerebral aneurysms results in significantly better packing than is achieved with helical coils of 0.010-inch-diameter wire. PMID- 15814942 TI - Changes in the intraaneurysmal pressure due to HydroCoil embolization. AB - BACKGROUND AND PURPOSE: The purpose of this study was to measure the changes in the intraaneurysmal fluid pressure and parent vessel flow characteristics resulting from packing the aneurysmal sac with hydrogel-coated coils. METHODS: Platinum coils coated with an expansible hydrogel were used to embolize a silicone model of the basilar tip aneurysm. The intraaneurysmal fluid pressure was measured with a NeuroCare Camino Pressure Micro-Probe at various packing ratios. A programmable pulsatile pump was used to input a physiologically relevant pulsatile flow in the parent artery. RESULTS: The intraaneurysmal fluid pressure did not increase when packing the aneurysm with hydrogel-coated platinum coils, even with a coil density up to 93%. CONCLUSION: Packing the aneurysm with hydrogel-coated coils at a density up to 93% did not increase the intraaneurysmal fluid pressure. PMID- 15814943 TI - Three-dimensional fusion digital subtraction angiography: new reconstruction algorithm for simultaneous three-dimensional rendering of osseous and vascular information obtained during rotational angiography. AB - This report describes three-dimensional (3D) fusion digital subtraction angiography (FDSA), a new algorithm for rotational angiography that combines reconstructions of the blood vessels and the osseous frame in a single 3D representation. 3D-FDSA is based on separate reconstructions of the mask and contrast sequences of the rotational acquisition. The two independent 3D data sets (3D-bone and 3D-digital subtraction angiography [DSA]) are fused in a single 3D representation. The algorithm uses a modification of the Feldkamp method that compensates for signal intensity inhomogeneity inherent to the reconstruction of nonsubtracted rotational acquisitions. By separately reconstructing the osseous and vascular information obtained from the rotational angiogram, 3D-FDSA provides optimal angiographic resolution and precise topographic analysis even when the studied vascular tree is in the immediate vicinity of bone. PMID- 15814944 TI - Angiographic evidence of aneurysm neck healing following endovascular treatment with bioactive coils. AB - A new angiographic finding in intracranial aneurysms embolized with Matrix coils is described. Two illustrative cases with a well-defined radiolucent separation between the coil mass and the parent artery are presented. To the best of our knowledge, this is the first report in humans of this finding. On the basis of prior histopathologic studies, this finding, which we call the "white-collar sign," may indicate the formation of a thick connective tissue barrier between aneurysm and artery that prevents any further aneurysmal filling. PMID- 15814945 TI - De novo basilar top aneurysm in an elderly patient treated with Guglielmi detachable coils. AB - Ruptured de novo aneurysms, compared with the usual subarachnoid hemorrhage, commonly occur in younger patients and are extremely rare in elderly patients. We discuss their etiology and report the case of a ruptured de novo aneurysm in a 77 year-old woman. PMID- 15814946 TI - Aneurysm regression after coil embolization of a concurrent aneurysm. AB - A 43-year-old woman with two incidental paraclinoid internal carotid artery aneurysms underwent coil embolization of the larger superior hypophyseal aneurysm and 10 weeks later underwent follow-up angiography that showed regression of the smaller, more distal paraclinoid aneurysm. We demonstrate that, although it is a rare occurrence, aneurysms can involute. We discuss potential mechanisms of this phenomenon and review the literature on aneurysm regression. PMID- 15814947 TI - Origins of the segmental arteries in the aorta: an anatomic study for selective catheterization with spinal arteriography. AB - BACKGROUND AND PURPOSE: The segmental arteries, which include the posterior intercostal, subcostal, and lumbar arteries, are gateways for performance of selective spinal arteriography of the thoracolumbar level. We performed a cadaveric study to clarify the anatomic relationship between the origins of the segmental arteries in the aorta and the vertebral column. METHODS: Five adult cadaveric aortas with intact thoracolumbar spines were dissected under magnification. In each specimen, nine pairs of posterior intercostal arteries, one pair of subcostal arteries, and four pairs of lumbar arteries were examined in detail. RESULTS: The origin of the posterior intercostal arteries in the upper thoracic level was situated at most about two levels caudal to the feeding level, whereas the origins in the lower levels were just caudal to the corresponding levels. The position of the bilateral orifices of the segmental arteries in the axial plane of the aorta was on the medial side at the thoracic levels, whereas it was on the dorsal side at the lumbar level. The horizontal distance between the orifices in the lumen of aorta was found to be wider at the thoracic level than the lumbar level, and the longitudinal distance was higher at the lower level, corresponding to the height of the vertebra. CONCLUSION: Understanding the 3D relationship of the initial segment of the segmental arteries with reference to the aorta and vertebral column is necessary for performance of the selective spinal arteriographic examination rationally by using a two-dimensional fluorescent display. PMID- 15814948 TI - N-butyl 2-cyanoacrylate embolization of spinal dural arteriovenous fistulae: CT evaluation, technical features, and outcome prognosis in 26 cases. AB - BACKGROUND AND PURPOSE: This study investigated the role of CT as an early predictor of outcome prognosis after glue embolization of spinal dural arteriovenous fistulas (SDAVF). METHODS: Over a 13-year period, 26 patients underwent glue embolization of SDAVF and were retrospectively reviewed. Immediately after embolization, each patient had CT evaluation of cast position. Mean follow-up angiography was 23.4 months (range, 1-87 months; median, 21 months). Both MR images and clinical data (e.g., gait and micturition disabilities according to the Aminoff and Logue disability scale, deep and superficial sensitivity) were analyzed, with a mean follow-up of 37.7 months (range, 12-98 months; median, 28 months). Data were tested by univariate analysis by using Fisher's exact test and the Kruskall Wallis test, depending on the order of the qualitative variables. RESULTS: Glue was found in the dura mater on CT in 19 (73%) patients. None of these patients had a recanalized fistula on angiography, and the clinical status of all of them improved. Glue was observed in or proximal to the foramen on CT in seven (27%) patients. In five of them, the fistula was recanalized within a mean period of 9 months (range, 3-24 months; median, 6 months) and their clinical status worsened. All five required surgical treatment. On angiography, the absence of fistula recanalization was observed in 21 (81%) patients and correlated with improvements in gait (P = .016), sensitivity (P = .030), and micturition (P = .080). It also correlated with a decrease in the extent of the abnormally high intramedullary T2 signal intensity (P = .002), a decrease in spinal cord diameter (P = .017), and the resolution of prominent perimedullary vessels (P < .001). The presence of glue within the dura mater on CT correlated with the absence of fistula recanalization (P = .045) and with overall improvement in clinical status, including gait and/or sensitivity and/or micturition (P = .042). CONCLUSION: CT evaluation of embolization cast position immediately after embolization may constitute an early and reliable tool for predicting permanent fistula occlusion and the prognosis for outcome. PMID- 15814949 TI - Hemangioblastomas of the lower spinal region: report of four cases with preoperative embolization and review of the literature. AB - BACKGROUND AND PURPOSE: Hemangioblastomas (HBs) are rare lesions accounting for 1 5% of all spinal cord tumors. Seventy-five percent of spinal HBs are intramedullary. Lesions of the conus medullaris and the cauda equina are uncommon, and the filum terminale location is very rare. HBs of the lower spinal region are highly vascular tumors requiring surgery that is potentially complicated by excessive bleeding. In the literature, there are few reports concerning preoperative embolization of HBs, and only few cases are reported in spinal location. Presurgical embolization of HBs located in the lower spinal region has not been described. Although lower spinal HBs are predominantly fed by the anterior spinal artery (ASA), embolization of these lesions is possible and can reduce tumor vascular supply, thus facilitating surgery. We report our experience in four rare cases of solitary HBs occurring in the lower spinal region. METHODS: Clinical charts and radiologic studies of four patients with a preoperatively embolized HB of the lower spinal region were retrospectively reviewed. The lesions were located in the conus medullaris in one case, at the level of the cauda equina in another, and in the filum terminale in two. In the conus medullaris case, the neoplasm was associated with a syrinx. In three patients, the HB was sporadic, while the patient with the HB of the cauda equina had Von Hippel-Lindau disease. Devascularization of the tumor was performed through the ASA in all cases and also through the posterior spinal artery in one by using non reabsorbable calibrated microspheres in three cases and polyvinyl alcohol particles in the other one. RESULTS: Embolization caused no permanent complications, although one patient with a cauda equina HB mildly worsened after the endovascular procedure but recovered before surgery. At surgery, the tumor was completely removed in all cases. Blood loss was reported to be less than usually observed (100, 200, 200, and 400 mL). In addition, manipulation and removal of the tumor was reported to be easy in three of four tumors. Histologic examination confirmed the diagnosis. At 1-year postsurgical follow-up, two patient recovered completely from neurologic deficits, and two showed significant recovery. No tumor recurred during a follow-up period of 1-6 years (mean, 3.5 years). CONCLUSION: Our results indicate that preoperative embolization of HBs of the lower spinal region is an useful procedure in aiding surgical resection of these highly vascular tumors. With a meticulous technique, embolization can be performed through the ASA. PMID- 15814950 TI - Normal cranial nerves in the cavernous sinuses: contrast-enhanced three dimensional constructive interference in the steady state MR imaging. AB - BACKGROUND AND PURPOSE: Three-dimensional (3D) constructive interference in steady state (CISS) MR imaging is useful for demonstrating cranial nerves (CNs) in the cistern. The purpose of this study was to evaluate normal CNs III, IV, V1, V2, and VI in the cavernous sinuses by using contrast-enhanced, three-dimensional (3D), Fourier transformation CISS MR imaging. METHODS: In 76 normal cavernous sinuses from 38 patients, detectability of CNs III-VI in the bilateral cavernous sinuses was evaluated by using contrast-enhanced 3D CISS MR imaging. In 40 cavernous sinuses from 20 patients, contrast-enhanced 3D CISS and contrast enhanced T1-weighted MR imaging were compared for the detectability of these CNs. RESULTS: Each CN was separately demonstrated, and in 11 patients (29%), all CNs in the cavernous sinuses were identified on contrast-enhanced 3D CISS MR imaging. The images depicted the intracavernous segments of CNs III, IV, V1, V2, and VI in 76 (100%), 46 (61%), 70 (92%), 67 (88%), and 73 (96%) of the 76 sinuses, respectively. In comparison of imaging techniques, contrast-enhanced 3D CISS MR imaging had a detection rate significantly higher than that of enhanced T1 weighting imaging (P < .05) in all CNs except for CN III, which was detected in 100% of cases with both techniques. CONCLUSION: Contrast-enhanced 3D CISS MR imaging provides clear images of each CN in the cavernous segment. This useful method may contribute to the diagnosis of diseases involving the cavernous sinuses, such as Tolosa-Hunt syndrome. PMID- 15814951 TI - Optic nerve diffusion measurement from diffusion-weighted imaging in optic neuritis. AB - BACKGROUND AND PURPOSE: Increases in apparent diffusion coefficient (ADC) from diffusion-weighted (DW) imaging are thought to be due to axonal disruption, and changes have been well documented in multiple sclerosis lesions. DW imaging of the optic nerves, however, presents many challenges. The goal of this study was to measure ADC in patients with optic neuritis by using zonal oblique multisection echoplanar imaging. METHODS: The optic nerves of eighteen patients who had experienced an attack of optic neuritis 1 year previously and 11 control subjects were imaged with the diffusion sequence (usable data were available from 16 patients and 10 control subjects). The orbital optic nerves were segmented by a blinded observer by using a computer-assisted threshold-based contouring technique, and the mean ADC was determined. RESULTS: The mean ADC from diseased optic nerves was 1324 x 10(-6) mm2/s, compared with 990 x 10(-6) mm2/s from healthy contralateral optic nerves (P = .005 versus diseased optic nerves) and 928 x 10(-6) mm2/s from control optic nerves (P = .006 versus diseased optic nerves and P = .40 versus healthy contralateral optic nerves). The diseased optic nerve ADC correlated with both visual (e.g., r(S) = 0.73; P = .001 for logMAR visual acuity) and electrophysiological parameters (e.g., r(S) = -0.57, P = .021 for visual evoked potential central field amplitude [VEP]). CONCLUSION: It has been possible to apply DW imaging in a patient population, and, in the chronic phase following optic neuritis, the correlation of mean ADC with the clinical and electrophysiological parameters suggests that the ADC is giving a surrogate measure of axonal disruption in the chronic, postinflammatory optic nerve lesion. PMID- 15814952 TI - Reappraisal of flow velocity ratio in common carotid artery to predict hemodynamic change in carotid stenosis. AB - BACKGROUND AND PURPOSE: Various Doppler criteria have been used to predict hemodynamically significant carotid stenosis. This study was performed to elucidate whether hemodynamically significant stenosis can be predicted indirectly by the blood flow velocity in the common carotid artery (CCA) measured with duplex ultrasonography in patients with unilateral stenosis of internal carotid artery (ICA). METHODS: Eighty-five patients who were scheduled to undergo carotid endarterectomy for unilateral stenosis of ICA origin were analyzed. The flow velocities and their side-to-side ratios in the CCA were calculated. The flow velocities in the CCA were measured with conventional ultrasonography and poststenotic blood flow with transoral carotid ultrasonography. Cerebral angiography was performed to evaluate the intracranial collateral flow. RESULTS: Among the absolute values and side-to-side ratios of Doppler flow velocities in the CCA, the end diastolic flow velocity (EDV) ratio in the CCA best correlated with the residual lumen area (r = 0.35; P = .0009), stenosis of diameter (r = 0.48; P < .0001), and poststenotic flow (r = 0.60; P < .0001). EDV ratios in the CCA were significantly lower in patients with collateral pathways (anterior communicating artery, P = .0005; posterior communicating artery, P = .004; ophthalmic artery, P < .0001; leptomeningeal collateral, P = .004). The optimal threshold value of the EDV ratio in the CCA for the presence of intracranial collateral flow and stenosis of diameter > or = 70% was 1.2. Those for tight stenosis in a cross-sectional area >95%, the reduction of poststenotic flow, and poststenotic narrowing were 1.4, 1.5, and 1.6, respectively. CONCLUSION: The EDV ratio in the CCA appears to be an additional parameter for predicting hemodynamically significant stenosis in patients with unilateral ICA stenosis. PMID- 15814953 TI - Hyrtl's fissure: a case of spontaneous cerebrospinal fluid otorrhea. AB - Spontaneous CSF otorrhea is a relatively rare entity. Adequate diagnosis and treatment are needed to avoid life-threatening complications such as meningitis. Because diagnosis is based on CT findings, identification of the different pathways of congenital fistulas requires detailed knowledge of embryology and anatomy. The facial canal, petromastoid canal, and tympanomeningeal (Hyrtl's) fissure can be responsible for CSF fluid otorrhea in the absence of any abnormality of the adjacent membranous labyrinth. We report the first documented and imaging case of Hyrtl's fissure and its treatment. PMID- 15814954 TI - Benign triton tumor of the trigeminal nerve. AB - Benign triton tumors (neuromuscular hamartomas) are rare neoplasms composed of well-differentiated striated muscle fibers admixed with peripheral nerve fibers. To the best of our knowledge, this is the third case reported in the world literature of a benign triton tumor involving cranial nerve V (trigeminal nerve) and the first in the radiology literature. The previous reports of this lesion have focused on its unusual pathology and not on the imaging findings. In light of the imaging appearance of this lesion, we consider that the imaging findings may suggest this uncommon diagnosis. PMID- 15814955 TI - Cricoarytenoid rheumatoid arthritis: an important consideration in aggressive lesions of the larynx. AB - The most frequent cause of an aggressive mass in the larynx is squamous cell carcinoma (SCC). Rheumatoid arthritis is known to affect the larynx but does not usually produce an aggressive mass. We present a case of rheumatoid arthritis in a 63-year-old woman who presented with acute upper airway obstruction. On CT scans, an erosive mass on the right cricoid cartilage with significant destruction of the surrounding structures was presumed to be an aggressive SCC. Surgical biopsies revealed rheumatoid arthritis of the cricoarytenoid joint. When a patient with rheumatoid arthritis presents with a mass in the larynx, cricoarytenoid rheumatoid arthritis should be ruled out even in the face of an aggressive lesion appearance at CT. PMID- 15814956 TI - Imaging of intraneural edema by using gadolinium-enhanced MR imaging: experimental compression injury. AB - BACKGROUND AND PURPOSE: Compressive and entrapment neuropathies are diseases frequently observed on routine clinical examination. A definitive diagnosis based on clinical symptoms and neurologic findings alone is difficult in many cases, however, and electrophysiologic measurement is used as a supplementary diagnostic method. In this study, we examined to use protein tracers (Evans blue albumin or horseradish peroxidase) and gadolinium-enhanced MR imaging to determine the changes of blood-nerve barrier permeability in compressive neuropathies. METHODS: In dogs, the median nerve was compressed for 1 hour by using five kinds of clips with various strengths (7.5-90-g force). After clip removal, the combined tracers of Evans blue albumin and gadolinium or horseradish peroxidase was administered intravenously as a tracer. After the animals were euthenized, we compared gadolinium-enhanced MR images with Evans blue albumin distribution in the nerve under fluorescence microscopy. The horseradish peroxidase-injected specimens were observed by transmission electron microscopy. RESULTS: On enhanced MR imaging, intraneural enhancement was caused by 60- and 90-g-force compression after 1 hour. Marked extravasation of protein tracers in the nerve occurred where there was compression by 60- and 90-g-force compression, and capillaries in the nerve showed the opening of tight junction and an increase of vesicular transport under the electron microscopy. This situation indicated breakdown of the blood-nerve barrier, with consequent edema formation and was seen as enhancement on MR imaging. CONCLUSION: Gadolinium-enhanced MR imaging can detect morphologic and functional changes of blood-nerve barrier in the nerve induced by mechanical compression. PMID- 15814957 TI - Is lumbar puncture contraindicated in patients with Chiari I malformation? PMID- 15814958 TI - Unusual fungal and pseudofungal infections of humans. PMID- 15814959 TI - Use of specific rRNA oligonucleotide probes for microscopic detection of Mycobacterium avium complex organisms in tissue. AB - Members of the Mycobacterium avium complex (MAC) are important environmental pathogens that are implicated in several chronic, idiopathic diseases. Diagnosis of MAC-based diseases is compromised by the need to cultivate these fastidious and slowly growing organisms in order to identify which mycobacterial species are present. Detection is particularly difficult when MAC is intracellular or embedded within mammalian tissues. We report on the development of culture independent, in situ hybridization (ISH) assays for the detection of MAC in culture, sputum, and tissue. This assay includes a highly reliable technique for the permeabilization of mycobacterial cells within culture and tissues. We describe a set of rRNA-based oligonucleotide probes that specifically detect either M. intracellulare, the two M. avium subspecies associated with human disease, or all members of MAC. The results call into question the validity of ISH results derived by the use of other gene loci, such as IS900. PMID- 15814960 TI - Identification of clinically relevant viridans streptococci by an oligonucleotide array. AB - Viridans streptococci (VS) are common etiologic agents of subacute infective endocarditis and are capable of causing a variety of pyogenic infections. Many species of VS are difficult to differentiate by phenotypic traits. An oligonucleotide array based on 16S-23S rRNA gene intergenic spacer (ITS) sequences was developed to identify 11 clinically relevant VS. These 11 species were Streptococcus anginosus, S. constellatus, S. gordonii, S. intermedius, S. mitis, S. mutans, S. oralis, S. parasanguinis, S. salivarius, S. sanguinis, and S. uberis. The method consisted of PCR amplification of the ITS regions by using a pair of universal primers, followed by hybridization of the digoxigenin-labeled PCR products to a panel of species-specific oligonucleotides immobilized on a nylon membrane. After 120 strains of the 11 species of VG and 91 strains of other bacteria were tested, the sensitivity and specificity of the oligonucleotide array were found to be 100% (120 of 120 strains) and 95.6% (87 of 91 strains), respectively. S. pneumoniae cross-hybridized to the probes used for the identification of S. mitis, and simple biochemical tests such as optochin susceptibility or bile solubility should be used to differentiate S. pneumoniae from S. mitis. In conclusion, identification of species of VS by use of the present oligonucleotide array is accurate and could be used as an alternative reliable method for species identification of strains of VS. PMID- 15814961 TI - por Variable-region typing by DNA probe hybridization is broadly applicable to epidemiologic studies of Neisseria gonorrhoeae. AB - The porin gene (porB) of Neisseria gonorrhoeae encodes the major outer membrane protein identified as PI or Por. To examine the utility of por variable-region (VR) typing, porB from 206 isolates was characterized by using oligonucleotide probes in a checkerboard hybridization assay that identifies the sequence types of five VRs of both PIA and PIB porB alleles. The strains represented temporally and geographically distinct isolates, isolates from a large cluster, epidemiologically linked partner isolates, and a collection of strains from disseminated gonococcal infections. By using rigorous epidemiologic criteria for transmission of infection between sex partners, por VR typing was more discriminatory than serovar typing in classifying isolates from both members of 43 epidemiologically linked pairs: 39 of 43 pairs were classified as coinciding by por VR typing compared to 43 of 43 by serovar determination (P = 0.058). porB sequence data confirmed the accuracy of the por VR method. Relationships between VR type and serovar typing monoclonal antibodies were observed for all six PIB and three of six PIA antibodies. por VR typing is a molecular tool that appears to have broad applicability. This method can be adapted to a wide range of technologies from simple hybridization to microarray and may allow for typing from noncultured clinical specimens. PMID- 15814962 TI - Evaluation of 16S rRNA sequencing and reevaluation of a short biochemical scheme for identification of clinically significant Bacteroides species. AB - Sequence analysis of the 16S rRNA gene represents a highly accurate and versatile method for bacterial classification and identification, even when the species in question is notoriously difficult to identify by phenotypic means. In this study, we evaluated the utility of 16S rRNA gene sequencing as a means of identifying clinically important Bacteroides species. We sequenced 231 clinical isolates that had been identified by a short biochemical scheme. Based on the sequence analysis, 192 clinical isolates were assigned to an established species, with the other 39 clinical strains revealing five unique sequences that may represent five novel species. This is in contrast to identification obtained from a short biochemical scheme, by which only 73.5% (172 of 231) of isolates were correctly identified to species level. Based on the solid identification obtained from 16S rRNA gene sequencing, the short biochemical scheme was modified and improved to provide clinical laboratories with an inexpensive and simple alternative for the identification of isolates of clinically significant Bacteroides species. PMID- 15814963 TI - Comparing genomes of Helicobacter pylori strains from the high-altitude desert of Ladakh, India. AB - The genomic diversity of Helicobacter pylori from the vast Indian subcontinent is largely unknown. We compared the genomes of 10 H. pylori strains from Ladakh, North India. Molecular analysis was carried out to identify rearrangements within and outside the cag pathogenicity island (cag PAI) and DNA sequence divergence in candidate genes. Analyses of virulence genes (such as the cag PAI as a whole, cagA, vacA, iceA, oipA, babB, and the plasticity cluster) revealed that H. pylori strains from Ladakh are genetically distinct and possibly less virulent than the isolates from East Asian countries, such as China and Japan. Phylogenetic analyses based on the cagA-glr motifs, enterobacterial repetitive intergenic consensus patterns, repetitive extragenic palindromic signatures, the glmM gene mutations, and several genomic markers representing fluorescent amplified fragment length polymorphisms revealed that Ladakhi strains share features of the Indo-European, as well as the East Asian, gene pools. However, the contribution of genetic features from the Indo-European gene pool was more prominent. PMID- 15814964 TI - Multilocus variable-number tandem repeat typing of Mycobacterium ulcerans. AB - The apparent genetic homogeneity of Mycobacterium ulcerans contributes to the poorly understood epidemiology of M. ulcerans infection. Here, we report the identification of variable number tandem repeat (VNTR) sequences as novel polymorphic elements in the genome of this species. A total of 19 potential VNTR loci identified in the closely related M. marinum genome sequence were screened in a collection of 23 M. ulcerans isolates, one Mycobacterium species referred to here as an intermediate species, and five M. marinum strains. Nine of the 19 loci were polymorphic in the three species (including the intermediate species) and revealed eight M. ulcerans and five M. marinum genotypes. The results from the VNTR analysis corroborated the genetic relationships of M. ulcerans isolates from various geographical origins, as defined by independent molecular markers. Although these results further highlight the extremely high clonal homogeneity within certain geographic regions, we report for the first time the discrimination of the two South American strains from Surinam and French Guyana. These findings support the potential of a VNTR-based genotyping method for strain discrimination within M. ulcerans and M. marinum. PMID- 15814966 TI - Comparison of an internally controlled, large-volume LightCycler assay for detection of Mycobacterium tuberculosis in clinical samples with the COBAS AMPLICOR assay. AB - We present a sensitive and specific assay for reliable and flexible detection of members of the Mycobacterium tuberculosis complex (MTBC) in clinical samples. This real-time PCR assay, which uses the LightCycler 2.0 instrument and 100-mul glass capillaries, can provide a result within 1 h after DNA extraction. The primers amplify a 206-bp fragment of the MTBC 16S rRNA gene. The sensor hybridization probe targets a region highly specific to members of the MTBC. The assay also includes a novel type of internal control that monitors the function of the reaction components and can detect potential inhibitors. Template DNA was extracted by the same procedure used for the COBAS AMPLICOR M. tuberculosis assay, so the LightCycler assay could be directly compared to the COBAS AMPLICOR assay. The LightCycler assay was evaluated with 146 clinical samples of various types. Very good agreement (100% sensitivity, 98.6% specificity) could be shown between the LightCycler and COBAS AMPLICOR assays. Specificity was checked with a panel of nontuberculous mycobacteria, as well as a large panel of bacterial and fungal organisms. PMID- 15814967 TI - European multicenter study of the LIAISON automated diagnostic system for determination of Toxoplasma gondii-specific immunoglobulin G (IgG) and IgM and the IgG avidity index. AB - The LIAISON system is a fully automated system based on chemiluminescence and antigen bound to magnetic microparticles. The system allows fast and precise measurement of Toxoplasma-specific immunoglobulin G (IgG) and IgM antibody levels and measurement of the IgG avidity index even at low levels of Toxoplasma specific IgG antibodies in a single step without manual interference. Seven European centers participated in a multicenter evaluation of the LIAISON system. The sensitivity and specificity of the LIAISON system compared to the Sabin Feldman dye test were 99.3 and 96.8%, respectively. In a comparison of the LIAISON Toxoplasma-specific IgM assay with an immunosorbent agglutination assay, the LIAISON assay had a sensitivity of 96.7% and a specificity of 95.4%. The LIAISON IgG assay showed agreements of 91, 100, and 100% with the AXSYM IgG (Abbott), VIDAS IgG (bioMerieux), and Platelia IgG (Bio-Rad) assays, respectively. The LIAISON IgM assay showed agreements of 95% with the AXSYM IgM and Platelia IgM assays, 96% with the ISAGA IgM assay (bioMerieux), and 97% with the VIDAS IgM assay. The coefficient of correlation between the LIAISON system and the VIDAS Toxoplasma-specific IgG avidity index was 0.81. By use of the Toxoplasma-specific IgG avidity index assay with specific IgM-positive samples, the diagnosis of infection with Toxoplasma gondii in early pregnancy has been improved significantly. The LIAISON avidity assay is a valuable assay for the exclusion of recently acquired infection with T. gondii (less than 4 months) in pregnant women, and it decreases significantly the necessity for follow-up testing. PMID- 15814965 TI - Genetic analysis of enteropathogenic and enterohemorrhagic Escherichia coli serogroup O103 strains by molecular typing of virulence and housekeeping genes and pulsed-field gel electrophoresis. AB - We investigated the genetic relationships of 54 Escherichia coli O103 strains from humans, animals, and meat by molecular typing of housekeeping and virulence genes and by pulsed-field gel electrophoresis (PFGE). Multilocus sequence typing (MLST) of seven housekeeping genes revealed seven profiles, I through VII. MLST profiles I plus III cover 45 Shiga toxin-producing E. coli (STEC) O103:H2 strains from Australia, Canada, France, Germany, and Northern Ireland that are characterized by the intimin (eae) epsilon gene and carry enterohemorrhagic E. coli (EHEC) virulence plasmids. MLST profile II groups five human and animal enteropathogenic E. coli (EPEC) O103:H2 strains that were positive for intimin (eae) beta. Although strains belonging to MLST groups II and I plus III are closely related to each other (92.6% identity), major differences were found in the housekeeping icdA gene and in the virulence-associated genes eae and escD. E. coli O103 strains with MLST patterns IV to VII are genetically distant from MLST I, II, and III strains, as are the non-O103 E. coli strains EDL933 (O157), MG1655 (K-12), and CFT073 (O6). Comparison of MLST results with those of PFGE and virulence typing demonstrated that E. coli O103 STEC and EPEC have recently acquired different virulence genes and DNA rearrangements, causing alterations in their PFGE patterns. PFGE typing was very useful for identification of genetically closely related subgroups among MLST I strains, such as Stx2 producing STEC O103 strains from patients with hemolytic uremic syndrome. Analysis of virulence genes contributed to grouping of E. coli O103 strains into EPEC and STEC. Novel virulence markers, such as efa (EHEC factor for adherence), paa (porcine adherence factor), and cif (cell cycle-inhibiting factor), were found widely associated with E. coli O103 EPEC and STEC strains. PMID- 15814968 TI - Clonal spread of a vancomycin-resistant Enterococcus faecium strain among bloodstream-infecting isolates in Italy. AB - Recent data indicated that the rate of vancomycin resistance in bloodstream infecting enterococcal isolates in Italy is one of the highest in Europe. The aims of this study were to characterize bloodstream-infecting vancomycin resistant enterococci (VRE) obtained from various Italian hospitals and to establish whether the isolates were clonally related. During the years 2001 to 2003, a total of 39 VRE isolates were obtained from 19 hospital laboratories in various areas of Italy. Species identification and resistance genotypes of the isolates were obtained by multiplex PCR. Further characterization included antibiotic susceptibility testing, pulsed-field gel electrophoresis (PFGE) of SmaI-digested genomic DNA, detection of virulence genes (esp and hyl), and multilocus sequence typing (MLST) of selected isolates. VRE were identified as 31 Enterococcus faecium (VREfm) isolates and 8 E. faecalis isolates. All but one isolate carried the vanA gene; one VREfm isolate carried the vanB gene. Analysis of the PFGE profiles showed that 28 VREfm isolates shared a similar electrophoretic profile, designed type 1, and were clonally related. All type 1 isolates were resistant to ampicillin, streptomycin, gentamicin, and rifampin and were positive for the esp gene. MLST identified an allelic profile (ST78) comprising purK allele 1, belonging to the C1 clonal lineage, characteristic of human infection and hospital outbreak isolates. The vanB-carrying VREfm isolate, of PFGE type 2, was shown to be a single-locus variant of ST78. Our data indicate that the recent increase in the number of bloodstream infections caused by VRE in Italy is due to the spread of a hospital-adapted, multidrug-resistant VREfm clone belonging to an internationally disseminated lineage. PMID- 15814969 TI - Loop-mediated isothermal amplification method targeting the lytA gene for detection of Streptococcus pneumoniae. AB - It is difficult to separate Streptococcus pneumoniae from the genotypically similar species Streptococcus mitis and Streptococcus oralis, which are commensals of the human oral cavity. A novel nucleic acid amplification technique, loop-mediated isothermal amplification (LAMP), which amplifies DNA under isothermal conditions (63 degrees C) with high specificity, efficiency, and rapidity, was examined regarding its applicability for detecting S. pneumoniae. An S. pneumoniae-specific LAMP primer targeting the lytA gene was designed. The primer specificity was validated using 10 Streptococcus and 7 non-Streptococcus species. Within 60 min, the assay could detect 10 or more copies of purified S. pneumoniae DNA with a sensitivity 1,000 times that of conventional PCR. Clinical isolates of 21 other strains (3 S. oralis, 17 S. mitis, and 1 Streptococcus species) that harbor virulence-factor-encoding genes (lytA or ply) were tried to differentiate S. pneumoniae. The detection of S. pneumoniae in clinical isolates was more selective using the LAMP method than using conventional PCR. Therefore, LAMP appears to be a sensitive and reliable means of diagnosing S. pneumoniae infection. PMID- 15814970 TI - Characterization of Borrelia lusitaniae isolates collected in Tunisia and Morocco. AB - Borrelia lusitaniae is a species within the complex Borrelia burgdorferi sensu lato and is infrequently isolated in Europe. In contrast, this species is by far the most predominant in North Africa and in Portugal. In this study, we analyzed the genetic diversity, at several loci, of a large population of isolates from free-living Ixodes ricinus ticks collected in Tunisia and Morocco. We found a moderate diversity of the whole genome by using pulsed-field gel electrophoresis as well as in the ospA gene sequences, compared to a high level of strain homogeneity in the small noncoding ribosomal spacer. In contrast, a high diversity of this locus has been previously reported for Portuguese isolates. We hypothesize that B. lusitaniae strains isolated in North Africa constitute a clone of Portuguese origin. PMID- 15814971 TI - Determinants for the occurrence of acute exacerbation of hepatitis B virus infection in Chinese patients after HBeAg seroclearance. AB - This study was performed to determine the factors for predicting the occurrence of acute exacerbation of hepatitis B virus infection in HBeAg-negative patients. Two hundred and sixteen patients with known times of HBeAg seroclearance were recruited. Liver biochemistry and virologic markers were monitored. Precore and core promoter mutations were determined by a line probe assay. The median age at HBeAg seroclearance was 34.5 years. The median follow-up duration was 26.4 months. Fifty-six (27.9%) patients had acute exacerbations. By Cox regression analysis, male gender, older age, and core promoter mutations at the time of HBeAg seroclearance were independently associated with the occurrence of acute exacerbation after HBeAg seroclearance (P = 0.025, 0.018, and 0.001, respectively). Fourteen (7.0%) patients had HBeAg seroreversion within a median follow-up period of 11.6 months after HBeAg seroclearance. By Cox regression analysis, older age at HBeAg seroclearance was independently associated with the chance of HBeAg seroreversion (P = 0.01). We concluded that male patients with core promoter mutations and delayed HBeAg seroclearance had a higher cumulative chance of acute exacerbation in the HBeAg-negative phase. Patients with delayed HBeAg seroclearance had a higher frequency of HBeAg seroreversion. PMID- 15814972 TI - Colonization of human immunodeficiency virus-infected outpatients in Taiwan with Candida species. AB - To understand the Candida colonization of human immunodeficiency virus (HIV) infected outpatients in Taiwan, we have conducted a prospective cohort study of Candida colonization and its risk factors at the National Taiwan University Hospital from 1999 to 2002. More than 50% of the patients were colonized with Candida species, and 12% developed symptomatic candidiasis. Patients colonized with fluconazole-resistant strains of Candida species had a higher prevalence of candidiasis than those colonized with susceptible strains. Our analysis found that antibiotic treatment and lower CD4(+) counts (<200 cells/mm(3)) increased the rate of oropharyngeal candidiasis in HIV-infected patients, while antiretroviral therapy protected patients from the development of candidiasis. PMID- 15814973 TI - Discrimination within phenotypically closely related definitive types of Salmonella enterica serovar typhimurium by the multiple amplification of phage locus typing technique. AB - Multilocus sequence typing (MLST) is a relatively new high-resolution typing system employed for epidemiological studies of bacteria, including Salmonella. Discrimination based on MLST of housekeeping genes may be problematical, due to the high identity of gene sequences of closely related Salmonella species. The presence of genomic sequences derived from stable temperate phages in Salmonella offers an alternative for MLST of Salmonella. We have used MLST of prophage loci in Salmonella enterica serovar Typhimurium to discriminate closely related isolates of serovar Typhimurium. We have compared these results to MLST of five housekeeping genes, as well as pulsed-field gel electrophoresis (PFGE). The presence or absence of prophage loci in the 73 serovar Typhimurium isolates tested, as well as allelic variation as detected by sequencing, provided greater discrimination between isolates than either MLST of housekeeping genes or PFGE. Amplification of prophage loci alone separated serovar Typhimurium isolates into 27 groups comprising multiple isolates or individual strains. Sequencing of isolates found within the clusters separated isolates even further. By contrast, PFGE could only divide the 73 isolates into five distinct groups. MLST using housekeeping genes did not provide any significant separation of isolates in comparison to amplification or MLST of prophage loci. The results demonstrate that the amplification and sequencing of prophage loci provides a high resolution, objective method for the discrimination of closely related isolates of serovar Typhimurium. It is proposed that multiple amplification of phage locus typing may provide sufficient discrimination for epidemiological purposes without recourse to MLST. PMID- 15814974 TI - Rapid detection of ofloxacin resistance in Mycobacterium tuberculosis by two low cost colorimetric methods: resazurin and nitrate reductase assays. AB - We have evaluated the performance of two rapid, low-cost methods for the detection of ofloxacin (OFX) resistance with 95 Mycobacterium tuberculosis isolates from countries with high multidrug-resistant tuberculosis endemicity. Results obtained by nitrate reductase and resazurin assays showed 100% agreement with those of the proportion method on 7H11 agar using 2 mug of OFX/ml. We confirmed the resistance of all isolates found to be resistant to OFX by the Mycobacterium Growth Indicator Tube system, and complete agreement among all methods was observed. Nitrate reductase and resazurin assays are rapid, simple, low-cost methods and might become inexpensive alternative procedures for rapid detection of OFX resistance in low-resource countries. PMID- 15814975 TI - Genetic diversity, determined on the basis of katG463 and gyrA95 polymorphisms, Spoligotyping, and IS6110 typing, of Mycobacterium tuberculosis complex isolates from Italy. AB - Mycobacterium tuberculosis complex isolates (n = 248) collected during a 1-year period in Tuscany, Italy, were genotyped for the katG463 and gyrA95 polymorphisms and by standard spacer oligonucleotide typing (spoligotyping) and IS6110 restriction fragment length polymorphism (RFLP) assays. Most of the isolates (n = 212; 85.5%) belonged to genotypic groups 2 and 3, which included most isolates from Italian-born patients. The remaining isolates were genotypic group 1 organisms, which were prevalent among foreign-born patients (29 of 36; 80.6%). Spoligotype analysis detected 116 unique patterns and 34 clusters including 166 isolates. The combination of spoligotyping and IS6110 RFLP analyses yielded 28 distinct clusters including 65 identical isolates (26.2%)--22 clusters with 2 isolates, 4 clusters with 3 isolates, 1 cluster with 4 isolates, and 1 cluster with 5 isolates--thus proving a low transmission rate in the community. Predominant spoligotypes representing 50% of clustered isolates were found in six clusters that included widespread type ST53 (clade T1) with 29 isolates (11.7% of total isolates); types ST50 and ST47 (Haarlem family) with 18 isolates (7.3%) and 8 isolates (3.2%), respectively; type ST42 (Latino-American and Mediterranean clade) with 13 isolates (5.2%); new type ST1737 (named "Tuscany") with 8 isolates (3.2%); and type ST1 (W-Beijing family) with 7 isolates (2.8%). Other spoligotype families, such as the Mycobacterium africanum, East African-Indian (EAI2/Manila), and central Asia 1 (CAS1/Delhi) families (all including organisms of genotypic group 1) and the Cameroun family (genotypic group 2), were detected especially among immigrant patients. The occurrence of genotypes originally found in distant geographic areas with a high prevalence of tuberculosis may represent a hallmark for changes in the dynamics of transmission of tuberculosis in the region in the near future. PMID- 15814976 TI - Rapid diagnostic method for detection of mumps virus genome by loop-mediated isothermal amplification. AB - Most mumps patients are clinically diagnosed without any virological examinations, but some diagnosed cases of mumps may be caused by other pathogens or secondary vaccine failure (SVF). To clarify these issues, a sensitive, specific, and rapid diagnostic method is required. We obtained 60 salivary swabs from 34 patients with natural infection during the course of the illness, 10 samples from patients with vaccine-associated parotitis, and 5 samples from patients with SVF. Total RNA was extracted and subjected to reverse transcription PCR (RT-PCR) and loop-mediated isothermal amplification (LAMP) for genome amplification. We detected mumps virus RNA corresponding to 0.1 PFU by LAMP within 60 min after RNA extraction, with the same sensitivity as RT-nested PCR. Mumps virus was isolated in 30 of 33 samples within day 2, and mumps virus genome was amplified by LAMP in 32 of them. The quantity of virus titer was calculated by monitoring the time to reach the threshold of turbidity. The viral load decreased after day 3 and was lower in patients serologically diagnosed as having SVF with milder illness. Accuracy of LAMP for the detection of mumps virus genome was confirmed; furthermore, it is of benefit for calculating the viral load, which reflects disease pathogenesis. PMID- 15814977 TI - Species-level identification of isolates of the Acinetobacter calcoaceticus Acinetobacter baumannii complex by sequence analysis of the 16S-23S rRNA gene spacer region. AB - The species Acinetobacter calcoaceticus, A. baumannii, genomic species 3, and genomic species 13TU included in the Acinetobacter calcoaceticus-Acinetobacter baumannii complex are genetically highly related and difficult to distinguish phenotypically. Except for A. calcoaceticus, they are all important nosocomial species. In the present study, the usefulness of the 16S-23S rRNA gene intergenic spacer (ITS) sequence for the differentiation of (genomic) species in the A. calcoaceticus-A. baumannii complex was evaluated. The ITSs of 11 reference strains of the complex and 17 strains of other (genomic) species of Acinetobacter were sequenced. The ITS lengths (607 to 638 bp) and sequences were highly conserved for strains within the A. calcoaceticus-A. baumannii complex. Intraspecies ITS sequence similarities ranged from 0.99 to 1.0, whereas interspecies similarities varied from 0.86 to 0.92. By using these criteria, 79 clinical isolates identified as A. calcoaceticus (18 isolates) or A. baumannii (61 isolates) with the API 20 NE system (bioMerieux Vitek, Marcy l'Etoile, France) were identified as A. baumannii (46 isolates), genomic species 3 (19 isolates), and genomic species 13TU (11 isolates) by ITS sequencing. An identification rate of 96.2% (76 of 79 isolates) was obtained by using ITS sequence analysis for identification of isolates in the A. calcoaceticus-A. baumannii complex, and the accuracy of the method was confirmed for a subset of strains by amplified rRNA gene restriction analysis and genomic DNA analysis by AFLP analysis by using libraries of profiles of reference strains. In conclusion, ITS sequence-based identification is reliable and provides a promising tool for elucidation of the clinical significance of the different species of the A. calcoaceticus-A. baumannii complex. PMID- 15814979 TI - Loop-mediated isothermal amplification for rapid detection of Newcastle disease virus. AB - We have evaluated a diagnostic system based on the loop-mediated isothermal amplification (LAMP) assay for the rapid, simple, and sensitive detection of Newcastle disease virus (NDV) directly from culture isolates as well as clinical samples. By using one set of specific primers targeting the fusion protein gene, the LAMP assay rapidly amplified the target gene within 2 h, requiring only a regular laboratory water bath or heat block for reaction. The results obtained from testing the genomes of 38 NDV strains, other different viruses, and clinical samples of experimentally infected chickens showed that LAMP was as sensitive and specific as nested PCR. All LAMP-positive samples were positive by nested PCR. The LAMP assay is faster than nested PCR, cost-effective, and easy to perform. Our results clearly demonstrate that the LAMP-based assay is a useful tool for the rapid and sensitive diagnosis of NDV infection. PMID- 15814978 TI - Clonal dissemination of macrolide-resistant and penicillin-susceptible serotype 3 and penicillin-resistant Taiwan 19F-14 and 23F-15 Streptococcus pneumoniae isolates in Japan: a pilot surveillance study. AB - Large-scale surveillance studies using molecular techniques such as pulsed-field gel electrophoresis (PFGE) have revealed that the spread of antibiotic-resistant pneumococci is due to clonal spread. However, in Japan, surveillance studies using such molecular techniques have never been done. Therefore, we conducted a pilot surveillance study to elucidate the present situation in Japan. Among the 145 isolates examined, the most prevalent serotype was type 19F (20%), for which most isolates were not susceptible to penicillin (86.2%) but were positive for the mef(A)/mef(E) gene (89.7%). The secondmost prevalent was serotype 3 (16.6%), for which most isolates were susceptible to penicillin (87.5%) and positive for the erm(B) gene (91.7%). PFGE analysis showed that both serotypes consisted mainly of clonally identical or related isolates and, in particular, 38% of the type 19F isolates were indistinguishable from or closely related to the Taiwan 19F-14 clone. In addition, some of the Japanese type 23F isolates with the erm(B) gene were indistinguishable from or related to the Taiwan 23F-15 clone as analyzed by PFGE. Based on the results of our pilot study performed in a single institution, it is likely that international antibiotic-resistant clones have already spread in Japan; therefore, a nationwide surveillance study should be urgently conducted. PMID- 15814980 TI - Laboratory-based surveillance and molecular epidemiology of influenza virus in Taiwan. AB - A laboratory-based surveillance network of 11 clinical virological laboratories for influenza viruses was established in Taiwan under the coordination of the Center for Disease Control and Prevention (CDC), Taiwan. From October 2000 to March 2004, 3,244 influenza viruses were isolated, including 1,969 influenza A and 1,275 influenza B viruses. The influenza infections usually occurred frequently in winter in the northern hemisphere. However, the influenza seasonality in Taiwan was not clear during the four seasons under investigation. For example, the influenza A viruses peaked during the winters of 2001, 2002, and 2003. However, some isolated peaks were also found in the summer and fall (June to November) of 2001 and 2002. An unusual peak of influenza B also occurred in the summer of 2002 (June to August). Phylogenetic analysis shows that influenza A isolates from the same year were often grouped together. However, influenza B isolates from the year 2002 clustered into different groups, and the data indicate that both B/Victoria/2/87-like and B/Yamagata/16/88-like lineages of influenza B viruses were cocirculating. Sequence comparison of epidemic strains versus vaccine strains shows that many vaccine-like Taiwanese strains were circulating at least 2 years before the vaccine strains were introduced. No clear seasonality of influenza reports in Taiwan occurred in contrast to other more continental regions. PMID- 15814981 TI - Efficient discrimination within a Corynebacterium diphtheriae epidemic clonal group by a novel macroarray-based method. AB - A large diphtheria epidemic in the 1990s in Russia and neighboring countries was caused by a clonal group of closely related Corynebacterium diphtheriae strains (ribotypes Sankt-Peterburg and Rossija). In the recently published complete genome sequence of C. diphtheriae strain NCTC13129, representative of the epidemic clone (A. M. Cerdeno-Tarraga et al., Nucleic Acids Res. 31:6516-6523, 2003), we identified in silico two direct repeat (DR) loci 39 kb downstream and 180 kb upstream of the oriC region, consisting of minisatellite (27- to 36-bp) alternating DRs and variable spacers. We designated these loci DRA and DRB, respectively. A reverse-hybridization macroarray-based method has been developed to study polymorphism (the presence or absence of 21 different spacers) in the larger DRB locus. We name it spoligotyping (spacer oligonucleotide typing), analogously to a similar method of Mycobacterium tuberculosis genotyping. The method was evaluated with 154 clinical strains of the C. diphtheriae epidemic clone from the St. Petersburg area in Russia from 1997 to 2002. By comparison with the international ribotype database (Institut Pasteur, Paris, France), these strains were previously identified as belonging to ribotypes Sankt-Peterburg (n = 79) and Rossija (n = 75). The 154 strains were subdivided into 34 spoligotypes: 14 unique strains and 20 types shared by 2 to 46 strains; the Hunter Gaston discriminatory index (HGDI) was 0.85. DRB locus-based spoligotyping allows fast and efficient discrimination within the C. diphtheriae epidemic clonal group and is applicable to both epidemiological investigations and phylogenetic reconstruction. The results are easy to interpret and can be presented and stored in a user-friendly digital database (Excel file), allowing rapid type determination of new strains. PMID- 15814982 TI - Dynamic range of hepatitis C virus RNA quantification with the Cobas Ampliprep Cobas Amplicor HCV Monitor v2.0 assay. AB - Accurate quantification of hepatitis C virus (HCV) RNA is needed in clinical practice to decide whether to continue or stop pegylated interferon-alpha ribavirin combination therapy at week 12 of treatment for patients with chronic hepatitis C. Currently the HCV RNA quantification assay most widely used worldwide is the Amplicor HCV Monitor v2.0 assay (Roche Molecular Systems, Pleasanton, Calif.). The HCV RNA extraction step can be automated in the Cobas Ampliprep device. In this work, we show that the dynamic range of HCV RNA quantification of the Cobas Ampliprep/Cobas Amplicor HCV Monitor v2.0 procedure is 600 to 200,000 HCV RNA IU/ml (2.8 to 5.3 log IU/ml), which does not cover the full range of HCV RNA levels in infected patients. Any sample containing more than 200,000 IU/ml (5.3 log IU/ml) must thus be retested after dilution for accurate quantification. These results emphasize the need for commercial HCV RNA quantification assays with a broader range of linear quantification, such as real time PCR-based assays. PMID- 15814983 TI - Utility of pooled urine specimens for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in men attending public sexually transmitted infection clinics in Mumbai, India, by PCR. AB - Pooling urogenital specimens for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by nucleic acid amplification tests is an attractive alternative to individual testing. As pooling can reduce the costs of testing as well as labor, it has been advocated for use in resource-poor settings. However, it has neither been widely adopted nor evaluated for use in developing countries. We evaluated the practical use of pooling first-catch urine (FCU) specimens for the detection of C. trachomatis and N. gonorrhoeae from 690 men in Mumbai, India, by PCR. FCU, urethral smears, and swabs were collected from men seen at two sexually transmitted infection (STI) clinics. All laboratory testing was done at the Lokmanya Tilak General Hospital. Gram stain smears and culture isolation for N. gonorrhoeae were performed. Each FCU was tested individually and in pools using the Roche Amplicor PCR for C. trachomatis and N. gonorrhoeae with an internal control for inhibition. Specimen pools consisted of aliquots from five consecutively processed FCUs combined into an amplification tube. An optical density reading of > or =0.20 indicated a pool for which subsequent testing of individual samples was required. Prevalence by PCR on single specimens was 2.2% (15/690) for C. trachomatis and 5.4% (37/690) for N. gonorrhoeae. Compared to individual FCU results, pooling for C. trachomatis and N. gonorrhoeae had an overall sensitivity of 96.1% (50/52). Specificity was 96.5% (83/86) in that three pools required single testing that failed to identify a positive specimen. Pooling missed two positive specimens, decreased the inhibition rate, and saved 50.3% of reagent costs. In this resource-limited setting, the use of pooling to detect C. trachomatis and N. gonorrhoeae by PCR proved to be a simple, accurate, and cost-effective procedure compared to individual testing. PMID- 15814984 TI - Invasive group A streptococcal disease in Alberta, Canada (2000 to 2002). AB - Invasive group A streptococcal (iGAS) disease was placed under surveillance in Alberta in August 1999. The purpose of this study was to determine the incidence rates of iGAS infections throughout Alberta over a 3-year period (2000 to 2002) and to better understand the epidemiology of iGAS in this province. There were a total of 441 cases of invasive GAS disease over the 3 years examined (average population over 3 years, 3,055,765) and 47 deaths. The incidence in Alberta was 5.0 (2000), 5.7 (2001), and 3.8 (2002) per 100,000. The two main metropolitan regions (Edmonton and Calgary) had the majority of iGAS disease cases (305 cases), producing incidence rates of 4.8 (Edmonton) and 6.9 (Calgary) in 2000, 6.9 (Edmonton) and 6.6 (Calgary) in 2001, and 4.1 (Edmonton) and 3.9 (Calgary) in 2002, as well as deaths attributable to GAS (31 deaths). The three most prevalent M types were M1 (71 cases), M3 (52 cases), and MPT2967 (44 cases). With respect to age, the highest incidence rates occurred in those less than 1 year old (11.7 per 100,000) and those 65 years or older (11.5 per 100,000). Varicella virus infection preceded iGAS disease in 25% of children 8 years of age and under. A seasonal association was observed during the 3 years studied, with the highest number of cases occurring in the winter months and the lowest occurring during the summer months. The data for years 2000 and 2001 show that the metropolitan regions of Alberta experienced some of the highest incidence rates reported in North America in the past decade. PMID- 15814985 TI - Identification of genotype 3 hepatitis E virus (HEV) in serum and fecal samples from pigs in Thailand and Mexico, where genotype 1 and 2 HEV strains are prevalent in the respective human populations. AB - Hepatitis E virus (HEV), the causative agent of hepatitis E, is an important public health concern in many developing countries. Increasing evidence indicates that hepatitis E is a zoonotic disease. There exist four major genotypes of HEV, and HEV isolates identified in samples from pigs belong to either genotype 3 or 4. Genotype 1 and 2 HEVs are found exclusively in humans. To determine whether genotype 1 and 2 HEVs also exist in pigs, a universal reverse transcription-PCR assay that is capable of detecting all four HEV genotypes was used to test for the presence of HEV RNA in serum and/or fecal samples from pigs in Thailand, where genotype 1 human HEV is prevalent, and from pigs in Mexico, where genotype 2 human HEV was epidemic. In Thailand, swine HEV RNA was detected in sera from 10/26 pigs of 2 to 4 months of age but not in sera from 50 pigs of other ages. In Mexico, swine HEV RNA was detected in 8/125 sera and 28/92 fecal samples from 2- to 4-month-old pigs. Antibodies to swine HEV were also detected in about 81% of the Mexican pigs. A total of 44 swine HEV isolates were sequenced for the open reading frame 2 gene region. Sequence analyses revealed that all swine HEV isolates identified in samples from pigs in Thailand and Mexico belong to genotype 3. Phylogenetic analyses revealed that minor branches associated with geographic origin exist among the swine HEV isolates. The results indicated that genotype 1 or 2 swine HEV does not exist in pigs from countries where the respective human HEV genotype 1 or 2 is prevalent. It is likely that only genotype 3 and 4 HEV strains have zoonotic potential. PMID- 15814986 TI - Identification and distribution of Acanthamoeba species genotypes associated with nonkeratitis infections. AB - Acanthamoeba is a free-living protozoan genus found in a wide variety of natural habitats, including water, soil, and air. Pathogenic isolates of Acanthamoeba are medically relevant as the causative agent of sight- threatening Acanthamoeba keratitis (AK), serious infections of other organs, and fatal granulomatous amebic encephalitis. Previous work employing DNA sequences of nuclear and mitochondrial small-subunit rRNA genes (SSU rRNA genes) determined the genotypic diversity of Acanthamoeba and found that many named species of Acanthamoeba are associated with particular genotypes. These studies also concluded that nearly all AK infections result from a single molecular genotype: T4. Here, we asked whether Acanthamoeba clinical isolates from non-AK infections are also associated with particular genotypes. DNA sequence determination of nuclear SSU rRNA genes was employed for genotypic identification of 29 isolates of Acanthamoeba from non AK infections. Sequence analysis demonstrates that T4 is the predominant genotype in non-AK infections, including those in brain, cerebrospinal fluid, nasal passages, skin, and lung. Rare genotypes (T1, T10, and T12) have been isolated from brain infections. We conclude that genotype T4 is the primary genotype in non-AK Acanthamoeba infections, as was the case in AK infections. However, the genotypes that were isolated from brains have not been observed in environmental isolates of Acanthamoeba, and their natural ecological niche is unknown. PMID- 15814987 TI - Analysis of the genetic structure of nontypeable pneumococcal strains isolated from conjunctiva. AB - More than 50% of the nontypeable (NT) pneumococcal strains received in our laboratory for reference purposes are isolated in sporadic cases of conjunctivitis. To determine the genetic structure of the population of these NT conjunctival strains, we analyzed 75 pneumococci (40 NT and 35 typeable) isolated from conjunctivas and 30 (15 NT and 15 typeable) isolated from other sources. The NT and typeable conjunctival strains grouped in separate clusters, whereas NT and typeable pneumococci isolated from other sources were similarly distributed. NT conjunctival strains belonged to two well-differentiated clonal lineages. The first, represented by three newly described sequence types, featured fully antibiotic susceptible strains and appeared to be characteristic of conjunctival tissue; the second, represented by the previously described ST344, had a pattern of multiresistance to penicillin, tetracycline, and erythromycin and shared a genetic background with some NT strains isolated from other sources. PMID- 15814988 TI - Use of variable-number tandem repeats to examine genetic diversity of Neisseria meningitidis. AB - Repetitive DNA motifs with potential variable-number tandem repeats (VNTR) were identified in the genome of Neisseria meningitidis and used to develop a typing method. A total of 146 meningococcal isolates recovered from carriers and patients were studied. These included 82 of the 107 N. meningitidis isolates previously used in the development of multilocus sequence typing (MLST), 45 isolates recovered from different counties in Norway in connection with local outbreaks, and 19 serogroup W135 isolates of sequence type 11 (ST-11), which were recovered in several parts of the world. The latter group comprised isolates related to the Hajj outbreak of 2000 and isolates recovered from outbreaks in Burkina Faso in 2001 and 2002. All isolates had been characterized previously by MLST or multilocus enzyme electrophoresis (MLEE). VNTR analysis showed that meningococcal isolates with similar MLST or MLEE types recovered from epidemiologically linked cases in a defined geographical area often presented similar VNTR patterns while isolates of the same MLST or MLEE types without an obvious epidemiological link showed variable VNTR patterns. Thus, VNTR analysis may be used for fine typing of meningococcal isolates after MLST or MLEE typing. The method might be especially valuable for differentiating among ST-11 strains, as shown by the VNTR analyses of serogroup W135 ST-11 meningococcal isolates recovered since the mid-1990s. PMID- 15814989 TI - Characterization of the genetic lineages responsible for pneumococcal invasive disease in Portugal. AB - The availability of a conjugate vaccine has the potential to reduce the disease burden of pneumococci and to alter the serotype frequency in the disease-causing population through immunoselection. These changes will probably be reflected in the distributions of individual genetic lineages within the population. We present a characterization of a collection of recent (1999 to 2002) invasive isolates from Portugal (n = 465) by macrorestriction profiling with pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. During this time, serotypes 14, 1, 3, 4, 8, 9V, 23F, 7F, 19A, and 12B were the 10 most prevalent overall by decreasing rank order. By combining the PFGE data with the sequence types (STs) of 104 isolates, we were able to identify the genetic lineages of the majority of the isolates. We found 66 STs, including 20 novel STs, corresponding to 47 different lineages by e-BURST analysis. We found in our collection a number of previously identified internationally disseminated lineages, especially among macrolide-resistant and penicillin-resistant isolates, and these accounted for most of the isolates. Most of the major lineages (17 of 25) were identified in all years of the study, suggesting that the pneumococcal population associated with invasive disease was stable. This study provides a characterization of the pneumococcal population associated with invasive disease that will be useful for detecting potential selective effects of the novel conjugate vaccine. PMID- 15814990 TI - Testing for induction of clindamycin resistance in erythromycin-resistant isolates of Staphylococcus aureus. AB - Disk diffusion and broth microdilution (BMD) were used to perform clindamycin (CLI) induction testing on 128 selected nonduplicate isolates of Staphylococcus aureus. Disk diffusion testing involved placing CLI and erythromycin (ERY) disks approximately 12 mm apart (measured edge to edge) on a Mueller-Hinton agar plate that had been inoculated with an S. aureus isolate; the plate was then incubated for 16 to 18 h. Two distinct induction phenotypes (labeled D and D(+)) and four noninduction phenotypes (designated as negative [Neg], hazy D zone [HD], resistant [R], and susceptible [S]) were observed in disk diffusion results. A clear, D-shaped zone of inhibition around the CLI disk was designated as the D phenotype and was observed for 21 isolates while a D-shaped zone containing inner colonies growing up to the CLI disk was designated as D(+) (17 isolates). In addition, 10 isolates were CLI susceptible and ERY resistant but were not inducible and showed no blunting of the CLI zone (Neg phenotype). Isolates that were CLI and ERY resistant (constitutive macrolide-lincosamide-streptogramin B resistance) demonstrated either a double zone of inhibition with an inner ring of reduced growth up to the edge of the disks (HD phenotype; 33 isolates) or solid growth around the CLI and ERY disks (R phenotype; 16 isolates). Finally, 31 isolates were susceptible by disk testing to both CLI and ERY (S phenotype). PCR results showed that isolates with a D phenotype harbored ermA, isolates with a D(+) phenotype contained either ermC (16 isolates) or ermA and ermC (one isolate), and all 10 isolates with a Neg phenotype contained msrA. All isolates with an HD or R phenotype harbored at least one erm gene. Isolates showing the D(+) phenotype by disk diffusion were also detected by BMD using a variety of CLI and ERY concentrations; however, isolates with the D phenotype were more difficult to detect by BMD and will likely require optimization of ERY and CLI concentrations in multilaboratory studies to ensure adequate sensitivity. Thus, at present, disk diffusion is the preferred method for testing S. aureus isolates for inducible CLI resistance. PMID- 15814991 TI - Usefulness of the hepatitis C virus core antigen assay for screening of a population undergoing routine medical checkup. AB - We studied the usefulness of the recently designed Trak-C assay for the detection and quantification of the hepatitis C virus (HCV) core antigen (Ag) for the screening of HCV infection in 4,201 subjects selected from 74,150 consecutive volunteers undergoing routine medical checkups. Subjects were selected for screening because they had risk factors (group II, n = 321) and/or elevated alanine transaminase activity (group I, n = 3847). Initially, the anti-HCV antibody assay and the Trak-C assay were performed on each patient. Subsequently, the Trak-C assay was performed only when the anti-HCV enzyme immune assay (EIA) was positive. Positive samples were further evaluated for anti-HCV antibodies by a third-generation strip immunoblot assay and for HCV RNA. Four samples (1.2%) from group II and 113 (2.9%) from group I were anti-HCV EIA positive. We also tested 33 subjects who previously tested positive for anti-HCV in our medical center. Among the 150 anti-HCV EIA-positive samples, the HCV core Ag result was in accord with the HCV RNA result in 146 cases (97.3%). When the EIA result was positive, the HCV core Ag concentration and the HCV RNA load were correlated (r(2) = 0.78; P < 0.001). Four samples with low viral loads were Trak-C negative but HCV RNA positive. Among the 2,395 anti-HCV EIA-negative serum samples collected during the first part of the study, 17 (0.7%) were found to contain very low levels of HCV core Ag (<8.5 pg/ml, the cutoff value being 1.5 pg/ml). All these samples were HCV RNA negative and considered to be false positives. This was confirmed by HCV core Ag neutralization analysis. The HCV core Ag assay is a useful method in the screening strategy of HCV infection and provides a reliable means of distinguishing between current and cleared HCV infections that is well correlated with HCV RNA testing. PMID- 15814992 TI - CHROMagar Candida medium for direct susceptibility testing of yeast from blood cultures. AB - An evaluation was performed on 95 blood cultures positive for Candida spp. to determine the correlation of direct susceptibility testing of fluconazole versus both standardized disk diffusion and MIC methods. For direct testing, an aliquot taken from BD BACTEC Plus and/or BD BACTEC Lytic/10 bottles (Becton Dickinson [BD], Sparks, MD) positive by gram stain for yeast was subcultured to CHROMagar Candida (BD), and a 25-microg fluconazole disk (BD) was placed on the plate. The area of growth inhibition surrounding the disk was measured at 24 and 48 h. In addition, a subculture of the isolate was tested by a microdilution MIC using YeastOne (TREK Diagnostics Systems Inc., OH) and disk diffusion (NCCLS M44-A) using a standardized inoculum plated onto CHROMagar Candida as well as Mueller Hinton agar to which 2% glucose and 0.5 microg/ml methylene blue dye was added (MH-GMB). The categorical interpretation derived from the MIC was used as the reference to which the disk diffusion results were compared. There were a total of 41 Candida albicans, 23 Candida glabrata, 20 Candida parapsilosis, 9 Candida tropicalis, and 1 each of Candida krusei and Candida lusitaniae tested. At 24 h there was full agreement among the methods for all C. albicans, C. tropicalis, C. lusitaniae, and C. krusei isolates. For the C. parapsilosis isolates at 24 h there was one very major discrepancy using the direct CHROMagar and one major error with the standardized MH-GMB. The majority of the errors were seen at 24 h with the C. glabrata isolates. Of the 23 C. glabrata isolates at 24 h by direct CHROMagar, there were 10 minor and 1 very major error; by MH-GMB there were 12 minor and 2 very major errors; and by standardized CHROMagar Candida there were 13 minor and 2 major errors. There were no very major errors with C. glabrata when all plates were read at 48 h. At 24 h by the direct and standardized CHROMagar the majority of C. glabrata isolates were more resistant, whereas by MH GMB they were more susceptible than the reference MIC interpretation. In summary, subculturing yeast directly from blood cultures onto CHROMagar to which a fluconazole disk has been added may provide a presumptive identification at 24 h and, with the exception of C. glabrata, was able to predict the susceptibility to fluconazole with the majority of Candida isolates examined in this evaluation. PMID- 15814993 TI - Phenotypic and genetic characterization of clinical isolates of CDC coryneform group A-3: proposal of a new species of Cellulomonas, Cellulomonas denverensis sp. nov. AB - CDC coryneform group A-3 bacteria are rare human pathogens. In this study, six group A-3 isolates (two from blood, one from cerebrospinal fluid, and one each from homograft valve, lip wound, and pilonidal cyst) were compared to the type strains of phenotypically related organisms, Cellulomonas fimi, Cellulomonas hominis, Oerskovia turbata, and Sanguibacter suarezii, and characterized by phenotypic, chemotaxonomic, and genotypic studies. DNA-DNA reassociation analysis identified two genomic groups, and phylogenetic analysis of the 16S rRNA gene sequence identified the taxonomic positions of these groups to genus level. Two groups were defined, and both were more closely related to Cellulomonas species: one group of three strains, for which we propose the new species Cellulomonas denverensis sp. nov., with the type strain W6929 (ATCC BAA-788(T) or DSM 15764(T)), was related to C. hominis ATCC 51964(T) (98.5% 16S rRNA gene sequence similarity), and the second group of three strains was related to C. hominis ATCC 51964(T) (99.8 to 99.9% 16S rRNA gene sequence similarity). The definition of this new Cellulomonas species and the confirmation of three strains as C. hominis serve to further clarify the complex taxonomy of CDC coryneform group A-3 bacteria and will assist in our understanding of the epidemiology and clinical significance of these microorganisms. PMID- 15814994 TI - Internally controlled real-time PCR monitoring of adenovirus DNA load in serum or plasma of transplant recipients. AB - Adenoviruses have been recognized as important pathogens in immunocompromised hosts. Particularly in pediatric allogeneic stem cell transplant recipients, the morbidity of the patients and mortality in those patients with disseminated infections have been found to increase over the last few years. Severe infections are predominantly but not exclusively caused by subgroup C adenoviruses. A multiplex real-time PCR assay using molecular beacons as probes was developed to enable monitoring of adenovirus DNA in those patients with simultaneous identification of subgroups. An internal control was coamplified in the multiplex PCR to check for the DNA isolation procedure as well as the presence of inhibitors in the clinical samples. The assay has been applied retrospectively in patient groups with different clinical outcomes of infection. In fatal cases, significantly higher adenovirus loads developed, exceeding even 10(11) copies/ml of serum or plasma. Patients with viral loads over 10(6) copies/ml appear to have an increased risk for fatal complications. This quantitative real-time PCR assay has been prospectively used clinically since 2002 to study the course of adenovirus infection. In addition, the assay provides objective start and end points of therapeutic interventions, including the clinically important evaluation of antiviral drugs. PMID- 15814995 TI - Use of an electronic nose to diagnose Mycobacterium bovis infection in badgers and cattle. AB - It is estimated that more than 50 million cattle are infected with Mycobacterium bovis worldwide, resulting in severe economic losses. Current diagnosis of tuberculosis (TB) in cattle relies on tuberculin skin testing, and when combined with the slaughter of test-positive animals, it has significantly reduced the incidence of bovine TB. The failure to eradicate bovine TB in Great Britain has been attributed in part to a reservoir of the infection in badgers (Meles meles). Accurate and reliable diagnosis of infection is the cornerstone of TB control. Bacteriological diagnosis has these characteristics, but only with samples collected postmortem. Unlike significant wild animal reservoirs of M. bovis that are considered pests in other countries, such as the brushtail possum (Trichosurus vulpecula) in New Zealand, the badger and its sett are protected under United Kingdom legislation (The Protection of Badgers Act 1992). Therefore, an accurate in vitro test for badgers is needed urgently to determine the extent of the reservoir of infection cheaply and without destroying badgers. For cattle, a rapid on-farm test to complement the existing tests (the skin test and gamma interferon assay) would be highly desirable. To this end, we have investigated the potential of an electronic nose (EN) to diagnose infection of cattle or badgers with M. bovis, using a serum sample. Samples were obtained from both experimentally infected badgers and cattle, as well as naturally infected badgers. Without exception, the EN was able to discriminate infected animals from controls as early as 3 weeks after infection with M. bovis, the earliest time point examined postchallenge. The EN approach described here is a straightforward alternative to conventional methods of TB diagnosis, and it offers considerable potential as a sensitive, rapid, and cost-effective means of diagnosing M. bovis infection in cattle and badgers. PMID- 15814996 TI - Species of Phaeoacremonium associated with infections in humans and environmental reservoirs in infected woody plants. AB - To date, three species of Phaeoacremonium have been associated with phaeohyphomycosis. These are P. parasiticum (formerly Phialophora parasitica), P. inflatipes, and P. rubrigenum. Numerous unknown isolates resembling Phaeoacremonium spp. have in recent years been isolated from human patients as well as from woody plants that appear to be the main environmental source of these fungi. Nine new Phaeoacremonium species, of which six were obtained as etiologic agents of human opportunistic infection, are reported. They can be identified based on their cultural and morphological characters, and the identifications are strongly supported in phylogenetic analyses of partial sequences of the actin, beta-tubulin, and calmodulin genes. A multiple-entry electronic key based on morphological, cultural, and beta-tubulin sequence data was developed to facilitate routine species identification. Reexamination of all isolates of P. inflatipes associated with human disease showed them to be misidentified and to belong to the new taxa described here. PMID- 15814997 TI - Evaluation of PCR testing of ethanol-fixed nasal swab specimens as an augmented surveillance strategy for influenza virus and adenovirus identification. AB - Viral culture isolation has been widely accepted as the "gold standard" for laboratory confirmation of viral infection; however, it requires ultralow temperature specimen storage. Storage of specimens in ethanol at room temperature could expand our ability to conduct active surveillance and retrospective screenings of viruses with rapid and inexpensive real-time PCR tests, including isolates from remote regions where freezing specimens for culture is not feasible. Molecular methods allow for rapid identification of viral pathogens without the need to maintain viability. We hypothesized that ethanol, while inactivating viruses, can preserve DNA and RNA for PCR-based methods. To evaluate the use of ethanol-stored specimens for augmenting surveillance for detection of influenza viruses A and B and adenoviruses (AdV), paired nasal swab specimens were collected from 384 recruits with febrile respiratory illness at Fort Jackson, S.C., in a 2-year study. One swab was stored at ambient temperature in 100% ethanol for up to 6 months, and the other swab was stored at -70 degrees C in viral medium. For viral detection, frozen specimens were cultured for a variety of respiratory viruses, and ethanol-fixed specimens were tested with TaqMan (TM) probe and LightCycler SYBR green (SG) melting curve assays with at least two different PCR targets for each virus. The sensitivities of the TM and SG assays on specimens stored in ethanol for 1 month were 75% and 58% for influenza A, 89% and 67% for influenza B, and 93 to 98% and 57% for AdV, respectively. Lower specificities of the real-time assays corresponded to the increased detection of PCR-positive but culture-negative specimens. Influenza virus RNA was detected as well or better after 6 months of storage in ethanol. PMID- 15814998 TI - Multilocus sequence typing versus pulsed-field gel electrophoresis for characterization of extended-spectrum beta-lactamase-producing Escherichia coli isolates. AB - Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains are emerging pathogens. Molecular typing of ESBL-producing E. coli is useful for surveillance purposes, to monitor outbreaks and track nosocomial spread. Although pulsed-field gel electrophoresis (PFGE) is the current "gold standard" for bacterial molecular typing, multilocus sequence typing (MLST) may offer advantages. Forty ESBL-producing E. coli isolates were selected at random from a cohort of intensive care unit patients who had active surveillance perirectal cultures done. PFGE identified 19 unique PFGE types (PT) among the 40 isolates; MLST identified 22 unique sequence types. MLST had greater discriminatory ability than PFGE for ESBL-producing E. coli. Simpson's indices of diversity for PFGE and MLST were 0.895 and 0.956, respectively. There were five clonal complexes (CCs) (isolates with differences of no more than two loci) that each contained multiple PT, but each PT was found in only one CC, indicating genetic consistency within a CC. MLST has clear utility in studies of ESBL-producing E. coli, based on a greater discriminatory ability and reproducibility than PFGE and the ability to a priori define genetically related bacterial strains. PMID- 15814999 TI - Detection and genotyping of Mycobacterium species from clinical isolates and specimens by oligonucleotide array. AB - Identification of pathogenic Mycobacterium species is important for a successful diagnosis of mycobacteriosis. The purpose of this study was to develop an oligonucleotide array which could detect and differentiate mycobacteria to the species level by using the internal transcribed spacer (ITS) sequence. Using a genus-specific probe and 20 species-specific probes including two M. avium intracellulare complex (MAC)-specific probes, we have developed an ITS-based oligonucleotide array for the rapid and reliable detection and discrimination of M. tuberculosis, MAC, M. fortuitum, M. chelonae, M. abscessus, M. kansasii, M. gordonae, M. scrofulaceum, M. szulgai, M. vaccae, M. xenopi, M. terrae, M. flavescens, M. smegmatis, M. malmoense, M. simiae, M. marinum, M. ulcerans, M. gastri, and M. leprae. All mycobacteria were hybridized with a genus-specific probe (PAN-03) for detection of the genus Mycobacterium. Mycobacterial species were expected to show a unique hybridization pattern with species-specific probes, except for M. marinum and M. ulcerans, which were not differentiated by ITS-based probe. Among the species-specific probes, two kinds of species-specific probes were designed for MAC in which there were many subspecies. The performance of the oligonucleotide array assay was demonstrated by using 46 reference strains, 149 clinical isolates, and 155 clinical specimens. The complete procedure (DNA extraction, PCR, DNA hybridization, and scanning) was carried out in 4.5 h. Our results indicated that the oligonucleotide array is useful for the identification and discrimination of mycobacteria from clinical isolates and specimens in an ordinary clinical laboratory. PMID- 15815000 TI - Reemergence of emm1 and a changed superantigen profile for group A streptococci causing invasive infections: results from a nationwide study. AB - Between 1999 and 2002, 496 invasive group A streptococcal (GAS) isolates from clinical microbiological departments in Denmark and subsequently 487 (98%) questionnaires from the clinicians treating the patients were received as part of a national surveillance. emm types and streptococcal superantigen (SAg) genes were determined. The incidence of invasive GAS infections was on average 2.3 per 100,000 per year. Bacteremia with no focal symptoms (27%) was together with erysipelas (20%) the most prevalent clinical diagnoses. Streptococcal toxic shock syndrome occurred in 10% of patients, of which 56% died. The overall case fatality rate within 30 days was 23%. In total, 47 different emm types were identified, of which emm1, emm3, emm4, emm12, emm28, and emm89 were identified in 72% of the 493 available isolates. During the 4-year period the presence of emm1 increased from 16% in 1999 to 40% in 2002. Concurrently, the presence of emm3 decreased from 23% in 1999 to 2% in 2002. The emm1 isolates predominantly carried speA, although the frequency decreased from 94% in 1999 to 71% in 2002, whereas the emm1-specific prevalence of speC increased from 25 to 53%. In a historical perspective, this could be interpreted as a reemergence of emm1 and could indicate a possible introduction of a new emm1 subclone. However, this reemergence did not result in any significant changes in the clinical manifestations during the study period. Our results show the complexity of invasive GAS infections, with time-dependent variations in the incidence and distribution of emm and SAg genes, which emphasizes the need for continuous epidemiological and molecular investigations. PMID- 15815001 TI - Analysis of Mycobacterium tuberculosis genotypes in Madrid and identification of two new families specific to Spain-related settings. AB - In Spain, tuberculosis (TB) patterns are changing because of the recent increase in the number of cases among immigrants. To establish the composition of circulating Mycobacterium tuberculosis strains before the effects of foreign strains appear, this study focused on molecular characterization of 233 patient isolates using spoligotyping. The spoligotyping data were further analyzed using an international database, SpolDB4. The results obtained showed that the general features of the M. tuberculosis population in Spain are coherent with those of other European countries, with the Latin American and Mediterranean group, and with the Haarlem 3 and T1 families as the most prevalent genotypes. The Spanish isolates clustered mostly with genotypes which had previously been isolated in countries linked with Spain. We also describe and fully characterize two novel M. tuberculosis families, Madrid1 and Madrid2, which are specific to Spain-related settings. The data reported here provide a solid reference when monitoring changes in the composition of the M. tuberculosis population in Spain as a consequence of the increasing rate of TB in the foreign population. PMID- 15815002 TI - System to assess genome sequencing needs for viral protein diagnostics and therapeutics. AB - Computational analyses of genome sequences may elucidate protein signatures unique to a target pathogen. We constructed a Protein Signature Pipeline to guide the selection of short peptide sequences to serve as targets for detection and therapeutics. In silico identification of good target peptides that are conserved among strains and unique compared to other species generates a list of peptides. These peptides may be developed in the laboratory as targets of antibody, peptide, and ligand binding for detection assays and therapeutics or as targets for vaccine development. In this paper, we assess how the amount of sequence data affects our ability to identify conserved, unique protein signature candidates. To determine the amount of sequence data required to select good protein signature candidates, we have built a computationally intensive system called the Sequencing Analysis Pipeline (SAP). The SAP performs thousands of Monte Carlo simulations, each calling the Protein Signature Pipeline, to assess how the amount of sequence data for a target organism affects the ability to predict peptide signature candidates. Viral species differ substantially in the number of genomes required to predict protein signature targets. Patterns do not appear based on genome structure. There are more protein than DNA signatures due to greater intraspecific conservation at the protein than at the nucleotide level. We conclude that it is necessary to use the SAP as a dynamic system to assess the need for continued sequencing for each species individually and to update predictions with each additional genome that is sequenced. PMID- 15815004 TI - Epidemiology and predictors of mortality in cases of Candida bloodstream infection: results from population-based surveillance, barcelona, Spain, from 2002 to 2003. AB - We conducted population-based surveillance for Candida bloodstream infections in Spain to determine its incidence, the extent of antifungal resistance, and risk factors for mortality. A case was defined as the first positive blood culture for any Candida spp. in a resident of Barcelona, from 1 January 2002 to 31 December 2003. We defined early mortality as occurring between days 3 to 7 after candidemia and late mortality as occurring between days 8 to 30. We detected 345 cases of candidemia, for an average annual incidence of 4.3 cases/100,000 population, 0.53 cases/1,000 hospital discharges, and 0.73 cases/10,000 patient days. Outpatients comprised 11% of the cases, and 89% had a central venous catheter (CVC) at diagnosis. Overall mortality was 44%. Candida albicans was the most frequent species (51% of cases), followed by Candida parapsilosis (23%), Candida tropicalis (10%), Candida glabrata (8%), Candida krusei (4%), and other species (3%). Twenty-four isolates (7%) had decreased susceptibility to fluconazole (MIC > or = 16 microg/ml). On multivariable analysis, early death was independently associated with hematological malignancy (odds ratio [OR], 3.5; 95% confidence interval [CI], 1.1 to 10.4). Treatment with antifungals (OR, 0.05; 95% CI, 0.01 to 0.2) and removal of CVCs (OR, 0.3; 95% CI, 0.1 to 0.9) were protective factors for early death. Receiving adequate treatment, defined as having CVCs removed and administration of an antifungal medication (OR, 0.2; 95% CI, 0.08 to 0.8), was associated with lower odds of late mortality; intubation (OR, 7.5; 95% CI, 2.6 to 21.1) was associated with higher odds. The incidence of candidemia and prevalence of fluconazole resistance are similar to other European countries, indicating that routine antifungal susceptibility testing is not warranted. Antifungal medication and catheter removal are critical in preventing mortality. PMID- 15815005 TI - Epidemiology of emerging methicillin-resistant Staphylococcus aureus (MRSA) in Denmark: a nationwide study in a country with low prevalence of MRSA infection. AB - Strict infection control measures introduced during the 1970s have kept the incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections extremely low in Denmark. Nevertheless, similarly to other countries, MRSA infections began to appear in the community in the late 1990s. A nationwide surveillance program has collected and stored all MRSA isolates since 1988 and, since 1999, clinical information has been also recorded. We used this information and isolates in a detailed epidemiological and molecular analysis of the 81 MRSA infections identified in Denmark in 2001. MRSA isolates were characterized by pulsed-field gel electrophoresis (PFGE), spa typing, multilocus sequence typing, and SCCmec typing. Comparison of the 45 community-onset MRSA (CO-MRSA) infections with the 36 hospital-acquired MRSA (HA-MRSA) infections showed several striking contrasts. Most CO-MRSA were recovered from skin and soft tissue infections caused by isolates carrying the Panton-Valentine leucocidin toxin genes, and the majority (84%) of isolates belonged to a single clonal type, ST80-IV, which has been found in the community in other European countries. Clone ST80-IV could be traced in Denmark back to 1993. ST80-IV was rarely found in HA-MRSA infections, which belonged to a large number of clonal types, including some pandemic MRSA clones. The low number of HA-MRSA infections and the diversity of MRSA clones in Danish hospitals may be the result of successful infection control measures that prevent spread of clones in hospitals. The mechanism of spread of the ST80-IV clone in the Danish community is not known, and new control measures are needed to control further spread of this and other CA-MRSA clones. PMID- 15815003 TI - Invasive infections caused by Trichosporon species and Geotrichum capitatum in patients with hematological malignancies: a retrospective multicenter study from Italy and review of the literature. AB - Trichosporonosis is an uncommon but frequently fatal mycosis in immunocompromised patients. A multicenter retrospective study was conducted to characterize cases of proven or probable invasive trichosporonosis diagnosed over the past 20 years in Italian patients with hematological diseases. Of the 52 cases identified, 17 were classified as Trichosporon sp. infections and 35 were attributed to Geotrichum capitatum. Acute myeloid leukemia accounted for 65.4% of the cases. The incidence rates of Trichosporon sp. and G. capitatum infections in acute leukemia patients were 0.4 and 0.5%, respectively. Overall, 76.9% of cases had positive blood cultures. Pulmonary involvement was documented in 26.9% of cases. Death was reported for 57.1% of G. capitatum infections and for 64.7% of Trichosporon sp. infections. A literature review on trichosporonosis in patients with any underlying disease or condition reveals G. capitatum as a predominantly European pathogen, particularly in certain Mediterranean areas, while Trichosporon sp. infections are seen with similar frequencies on all continents. The majority of published Trichosporon sp. and G. capitatum infections occurred in patients with hematological diseases (62.8 and 91.7%, respectively). Well over half of these were suffering from acute leukemia (68 and 84% of patients with Trichosporon sp. and G. capitatum infections, respectively). Crude mortality rates were 77% for Trichosporon spp. and 55.7% for G. capitatum. The optimal therapy for trichosporonosis has yet to be identified; however, in vitro experiences are providing encouraging evidence of the potential role of the new triazoles, in particular, voriconazole. PMID- 15815006 TI - Sequencing and resolution of amplified herpes simplex virus DNA with intermediate melting curves as genotype 1 or 2 by LightCycler PCR assay. AB - DNA from 101 specimens containing herpes simplex virus (HSV) produced atypical intermediate melting curves compared with those expected for HSV type 1 or HSV type 2 subsequent to real-time PCR. Nucleic acid sequence analysis of amplified target DNA revealed 1- or 3-bp polymorphisms in the probe region which allowed designation of these viruses as HSV genotype 1 or HSV genotype 2. These two subpopulations of HSV were also identified as HSV genotype 1 or HSV genotype 2 using another commercially available PCR method. Amplified HSV target DNA producing intermediate melting curves could be designated as HSV genotype 1 or HSV genotype 2 without performing sequencing or another PCR method with 96/101 (95%) specimens by adding known intermediate HSV DNA characteristic for the two subpopulations as controls. PMID- 15815007 TI - Evaluation and updating of the Osiris expert system for identification of Escherichia coli beta-lactam resistance phenotypes. AB - Osiris is a video zone size reader for disk diffusion tests that includes a built in extended expert system (EES). We evaluated the efficacy of the Osiris EES for the identification of beta-lactam susceptibility phenotypes of Escherichia coli isolates. Fifteen beta-lactam agents and three beta-lactam-beta-lactamase inhibitor combinations were tested by the disk diffusion method against 50 E. coli strains with well-characterized resistance mechanisms. The strains were screened for the production of extended-spectrum beta-lactamase (ESBL) by the double-disk synergy test using a disk of amoxicillin-clavulanic acid with disks of the extended-spectrum cephalosporins and aztreonam. Overall, the EES accurately identified the phenotype for 78% of the strains, indicated an inexact phenotype for 17% of the strains, and could not find a matching phenotype for the remaining 5% of the strains. The percentage of correct identification for each resistance mechanism was 100% for inhibitor-resistant TEM and for TEM plus cephalosporinase, 88.9% for TEM and for ESBL, 70.8% for cephalosporinase overproduction, and 25% for oxacillinase. The main cause of discrepancy was the misidentification of oxacillinase as inhibitor-resistant TEM. The conventional double-disk synergy test failed to detect ESBL production in two strains (one producing VEB-1 and one producing CTX-M-14), but synergy between cefepime and amoxicillin-clavulanic acid was visible after the distance between the disks was reduced to 20 mm. After the interpretative guidelines of the EES were updated according to our results, the percentage of correct phenotype identification increased from 78 to 96%. PMID- 15815008 TI - Detection of human immunodeficiency virus type 1 DNA in dried blood spots by a duplex real-time PCR assay. AB - A dried blood spot (DBS) is a well-accepted means for the collection, transport, and storage of blood samples for various epidemiologic, serologic, and molecular assays for human immunodeficiency virus (HIV) studies. It is particularly important for mother-to-infant-transmission studies of affected individuals living in remote areas. We have developed a real-time PCR method to detect HIV type 1 (HIV-1) DNA in dried blood spots. A cellular gene, RNase P, was coamplified with the HIV-1 DNA in the same tube to monitor the DNA extraction efficiency and the overall assay performance. Our assay is a one-tube, single step closed-system assay and uses a dUTP/uracil DNA glycosidase anti-PCR contamination control. The HIV-1 primers and probe were derived from a conserved region of the long terminal repeat. The detection of RNase P is attenuated by lowering the forward and reverse primer concentrations so that its amplification will not overwhelm the HIV-1 amplification and yet will provide a semiquantitative measurement of the quality of the isolated DBS DNA. We examined 103 HIV-1-seropositive and 56 seronegative U.S. adults and found that our assay has a sensitivity of 98.1% (95% confidence interval [CI], 95.5% to 100%) and specificity of 100% (95% CI, 99% to 100%). The positive and negative predictive values are 100% and 96.6%, respectively. This duplex PCR assay may be useful in identifying HIV-1-infected persons, particularly infants born to seropositive mothers in remote areas of the world. PMID- 15815009 TI - Use of quantitative real-time PCR to monitor the response of Chlamydophila felis infection to doxycycline treatment. AB - Fifteen cats infected with Chlamydophila felis were monitored for the presence of C. felis DNA on ocular swabs by using real-time PCR and for clinical signs of disease. The cats were assigned to three groups: oral doxycycline at 10 mg/kg of body weight/day for 7 days (six cats), oral doxycycline at 10 mg/kg/day for 14 days (five cats), and an untreated control group (four cats). The untreated cats remained positive for C. felis throughout the trial; clinical signs were most severe on days 14 to 21 postinfection, and then they declined. Treatment with 7 and 14 days of doxycycline decreased C. felis relative copy numbers and clinical signs rapidly. C. felis became undetectable in some of the cats during or after treatment. However, after the cessation of treatment, a recurrence of high relative copy numbers of C. felis and severe clinical signs in all cats was seen. Rescue treatment with 21 days of doxycycline was successful at eliminating infection in eight of the cats; a further 28 days of doxycycline was required to eliminate infection in the remaining three cats. It was concluded that 7, 14, and, in some cases, 21 days of treatment with oral doxycycline will not eliminate C. felis infection. At least 28 days of treatment with doxycycline is required to ensure elimination of the organism. Real-time PCR is a sensitive technique for monitoring C. felis infection and the response to antibiotic treatment. PMID- 15815010 TI - Molecular evidence for multiple host-specific strains in the genus Rhinosporidium. AB - The taxonomic relationship of Rhinosporidium seeberi with other organisms remained controversial for over a century. Recently, molecular studies have shown R. seeberi to be a protistal microbe in the newly described class Mesomycetozoea at the animal-fungal boundary. Phylogenetic analyses of R. seeberi using 18S small-subunit (SSU) rRNA genes from several hosts suggested Rhinosporidium as a monotypic genus. To test this hypothesis, the internal transcribed spacer 1 (ITS1), 5.8S, and ITS2 from eight humans, two swans, and a dog with rhinosporidiosis were sequenced. The ITS regions were amplified by PCR using a primer designed from a unique region of R. seeberi's 18S SSU rRNA genes in combination with the ITS4 universal primer. In addition, the universal ITS4 and ITS5 primers were also used. R. seeberi's ITS sequences showed differences in the numbers of nucleotides among strains. For instance, the eight human ITS sequences were uniformly similar with only a few mismatches and approximately 1,060 bp long. In contrast, sequences from one of the swans and the dog were 1,356 bp and approximately 1,147 bp long, respectively. Clustal analysis of all of the ITS sequences showed multiple 50- to 60-bp gaps and several mismatches among them. Parsimony analysis placed the Rhinosporidium ITS sequences in three well supported sister groups according to the hosts' identities. This analysis strongly suggests that the genus Rhinosporidium may possess multiple host specific strains. No correlation was found between this finding and the phenotypic features of R. seeberi in the studied samples. PMID- 15815012 TI - Borrelia burgdorferi ospC heterogeneity among human and murine isolates from a defined region of northern Maryland and southern Pennsylvania: lack of correlation with invasive and noninvasive genotypes. AB - B. burgdorferi invasiveness correlates with ospC genotype. To test this hypothesis and whether identical genotypes infect humans and small mammals in specific sites, B. burgdorferi ospC heterogeneity was tested among isolates from northern Maryland and southern Pennsylvania. Six culture-positive patients allowed collection of small animals from their properties, and spirochetes from animals trapped within 300 yards of each patient's home were isolated. 3' ospC sequences were compared to reference sequences. Of the 7 human and 15 mouse DNA templates that produced reliable sequences, all clustered with references into only four and seven distinct clades, respectively. A human and a mouse isolate with the same ospC were seen in only one locality, and five of six sites contained two or more B. burgdorferi ospC clones. Four invasive patient isolates and six small mammal isolates clustered with "noninvasive" reference ospC genotypes. A high degree of ospC diversity exists among B. burgdorferi isolates in Maryland and Pennsylvania, even in narrowly defined geographic localities. Dissemination in mice and humans by noninvasive ospC types contradicts the ospC invasiveness hypothesis. Alternative genetic markers for B. burgdorferi disseminated disease should be investigated. PMID- 15815013 TI - Nucleic acid amplification assays for detection of La Crosse virus RNA. AB - We report the development of nucleic acid sequence-based amplification (NASBA) and quantitative real-time reverse transcription (RT)-PCR assays for the detection of La Crosse (LAC) virus in field-collected vector mosquito samples and human clinical samples. The sensitivities of these assays were compared to that of a standard plaque assay in Vero cells. The NASBA and quantitative real-time RT PCR assays demonstrated sensitivities greater than that of the standard plaque assay. The specificities of these assays were determined by testing a battery of reference strain viruses, including representative strains of LAC virus and other arthropod-borne viruses. Additionally, these assays were used to detect LAC viral RNA in mosquito pool samples and human brain tissue samples and yielded results within less than 4 h. The NASBA and quantitative real-time RT-PCR assays detect LAC viral RNA in a sensitive, specific, and rapid manner; these findings support the use of these assays in surveillance and diagnostic laboratory systems. PMID- 15815011 TI - Molecular identification of mumps virus genotypes from clinical samples: standardized method of analysis. AB - A sensitive nested reverse transcription-PCR assay, targeting a short fragment of the gene encoding the small hydrophobic protein (SH gene), was developed to allow rapid characterization of mumps virus in clinical samples. The sensitivity and specificity of the assay were established using representative genotypes A, B, C, D, E, and F. Mumps virus RNA was characterized directly from cerebrospinal fluid (CSF) samples and in extracts of mumps virus isolates from patients with various clinical syndromes. Direct sequencing of products and subsequent phylogenetic analysis enabled genetic classification. A simple web-based system of sequence analysis was established. The study also allowed characterization of mumps virus strains from Argentina as part of a new subgenotype. This PCR assay for characterization of mumps infections coupled to a web-based analytical program provides a rapid method for identification of known and novel strains. PMID- 15815014 TI - Clinical manifestations of staphylococcal scalded-skin syndrome depend on serotypes of exfoliative toxins. AB - We sought a possible correlation between the clinical manifestations of staphylococcal scalded-skin syndrome (SSSS) and the serotype of exfoliative toxins (ET) by PCR screening of the eta and etb genes in Staphylococcus aureus strains isolated from 103 patients with generalized SSSS and 95 patients with bullous impetigo. The eta gene and the etb gene were detected in, respectively, 31 (30%) and 20 (19%) episodes of generalized SSSS and 57 (60%) and 5 (5%) episodes of bullous impetigo. Both genes were detected in 52 (50%) episodes of generalized SSS and 33 (35%) episodes of bullous impetigo. To explain this link between etb and generalized SSSS, we examined the distribution of ETA- and ETB specific antibodies in the healthy population (n = 175) and found that the anti ETB antibody titer was lower than the anti-ETA titer. Thus, ETA is associated with bullous impetigo and ETB is associated with generalized SSSS, possibly owing to a lower titer of anti-ETB neutralizing antibodies in the general population. PMID- 15815015 TI - Molecular identification of Capnocytophaga spp. via 16S rRNA PCR-restriction fragment length polymorphism analysis. AB - Capnocytophaga spp. have been implicated as putative periodontal pathogens associated with various periodontal diseases. Although the genus is known to contain five human oral isolates, accurate identification to species level of these organisms recovered from subgingival plaque has been hampered by the lack of a reliable method. Hence, most studies to date have reported these isolates as Capnocytophaga spp. Previous attempts at identification were based on biochemical tests; however, the results were inconclusive. Considering the differing virulence features of the respective isolates, it is crucial to identify these isolates to species level. The universal and conservative nature of the 16S rRNA gene has provided an accurate method for bacterial identification. The aim of this study was to identify Capnocytophaga spp. via restriction enzyme analysis of this gene (16S rRNA PCR-restriction fragment length polymorphism). The results (backed up by 16S rRNA gene sequencing) showed that this method reliably identifies all named Capnocytophaga spp. to species level. PMID- 15815016 TI - Molecular epidemiology of hepatitis C virus genotype 4 isolates in Egypt and analysis of the variability of envelope proteins E1 and E2 in patients with chronic hepatitis. AB - We analyzed hepatitis C virus (HCV) genotype 4 isolates circulating in the Alexandria District (Egypt) in terms of genetic divergence and the presence of different subtypes. Hypervariable region 1 (HVR1) and the NH2 region of the E2 protein were characterized, and the heterogeneity of subtype 4a isolates was evaluated by analyzing epitope frequencies, immunoproteasome prediction, and possible glycosylation patterns. The heterogeneity of the nucleotide sequences was greater than that found in previous studies, which reported only subtype 4a. Subtype 4a was most common (78% of cases), yet four new subtypes were found, with subtype 4m representing 11% of the cases and the other three subtypes representing another 11%. Substantial heterogeneity was also found when the intrasubtype 4a sequences were analyzed. Differences in the probability of glycosylation and in the positions of the different sites were also observed. The analysis of the predicted cytotoxic-T-lymphocyte epitopes showed differences in both the potential proteosome cleavage and the prediction score. The Egyptian isolates in our study also showed high variability in terms of the HVR1 neutralization epitope. Five of these isolates showed amino acid substitutions never previously observed (a total of six positions). Four of these residues (in four different isolates) were in positions involved in anchoring to the E2 glycoprotein core and in maintaining the HVR1 conformation. The results of this study indicate that HCV genotype 4 in Egypt is extremely variable, not only in terms of sequence, but also in terms of functional and immunological determinants. These data should be taken into account in planning the development of vaccine trials in Egypt. PMID- 15815017 TI - env Gene typing of human immunodeficiency virus type 1 strains on electronic microarrays. AB - The NanoChip system was used for subtyping human immunodeficiency virus type 1 (HIV-1) strains using probes complementary to the V1 region of the env gene. Probes for six subtypes (A to D, F, and G) and two circulating recombinant forms (AG and AE) of HIV-1 group M were included. The specificity of these oligonucleotides had been evaluated previously in a DNA enzyme immunoassay. Samples from 112 patient sera were used as templates in a nested reverse transcription-PCR to produce amplicons that were applied to the array. The array was then hybridized successively to pairs of oligonucleotide probes. The strains were assigned a subtype on the basis of their probe hybridization patterns. One strain gave a contradictory pattern and was designated as untypeable by the NanoChip assay. Eighty-eight strains gave hybridization patterns that allowed a correct subtype designation to be made by the NanoChip assay compared to either the sequence or the heteroduplex mobility assay (HMA)-determined subtypes. Thirteen strains that reacted with the subtype A probe (SA2) were incorrectly assigned to subtype A, or to one of the related circulating recombinant types (AE or AG), on the basis of reactions with probe SAE1 or SAG1. The results indicate that these oligonucleotides have relatively low specificities. The probe subtypes of three strains matched the subtypes determined for the gag and pol genes but not the env gene, suggesting that a recombination event may have occurred within the env gene. Overall, the NanoChip assay gave results comparable to those for HMA and sequencing and provides a convenient and cost-effective means by which to subtype HIV-1. PMID- 15815018 TI - Evaluation of broth microdilution antifungal susceptibility testing conditions for Trichophyton rubrum. AB - Fifty clinical isolates of Trichophyton rubrum were selected to test with ketoconazole, fluconazole, itraconazole, griseofulvin, and terbinafine by following the National Committee for Clinical Laboratory Standards susceptibility testing guidelines for filamentous fungi (M38-A). In addition, other susceptibility testing conditions were evaluated: (i) three medium formulations including RPMI 1640 (standard medium), McVeigh & Morton (MVM), and Sabouraud dextrose broth (SDB); (ii) two incubation temperatures (28 and 35 degrees C); and (iii) three incubation periods (4, 7, and 10 days). The strains Candida parapsilosis (ATCC 22019), Candida krusei (ATCC 6258), T. rubrum (ATCC 40051), and Trichophyton mentagrophytes (ATCC 40004) were included as quality controls. All isolates produced clearly detectable growth only after 7 days of incubation. MICs were significantly independent of the incubation temperature (28 or 35 degrees C) (P < 0.05). Different incubation periods resulted in MICs which were consistently different for each medium when azoles and griseofulvin were tested (P < 0.05). MICs obtained from different media at the same incubation time for the same isolate were significantly different when azoles and griseofulvin were tested (P < 0.05). MICs were consistently higher (usually 1 to 2 dilutions) with RPMI than with MVM or SDB (P < 0.05). When terbinafine was tested, no parameter had any influence on MICs (P < 0.05). RPMI standard medium appears to be a suitable testing medium for determining the MICs for T. rubrum. MICs obtained at different incubation times need to be correlated with clinical outcome to demonstrate which time has better reliability. PMID- 15815019 TI - Direct detection of Nocardia spp. in clinical samples by a rapid molecular method. AB - We developed a 16S PCR-based assay for the rapid detection of Nocardia spp. directly from human clinical samples. The applicability of the assay was confirmed by using 18 samples from patients with nocardiosis as diagnosed by conventional cultures and 20 clinical samples from patients with confirmed tuberculosis used as negative controls. PMID- 15815020 TI - Performance of CHROMagar MRSA medium for detection of methicillin-resistant Staphylococcus aureus. AB - CHROMagar MRSA was evaluated for its ability to identify methicillin-resistant Staphylococcus aureus (MRSA). A well-defined collection consisting of 216 MRSA strains and 241 methicillin-susceptible Staphylococcus aureus isolates was used. The sensitivity of CHROMagar MRSA after 24 h of incubation was 95.4%, increasing to 100% after 48 h. The specificity was already 100% after 24 h. PMID- 15815021 TI - Misidentification of Mycobacterium leprae as Mycobacterium intracellulare by the COBAS AMPLICOR M. intracellulare test. AB - Commercially available nucleic acid probe- and amplification-based systems for detection and differentiation of mycobacteria are widely used in clinical microbiology laboratories. Here we report two cases of human leprosy in which the COBAS AMPLICOR Mycobacterium intracellulare test led to false- positive results. Correct identification of Mycobacterium leprae was possible only by amplification and comparative sequence analysis of the 16S rRNA gene. PMID- 15815022 TI - Susceptibilities of Mycobacterium tuberculosis to isoniazid and rifampin on blood agar. AB - In this study, blood agar was used instead of 7H10 agar for the susceptibility testing of 34 clinical isolates of Mycobacterium tuberculosis to isoniazid (INH) and rifampin (RIF) in accordance with the NCCLS. The BACTEC 460 TB system (Becton Dickinson, Sparks, Md.) was used as a "gold standard." Results for both media were in agreement for RIF and INH at 100 and 94.1%, respectively. For INH, the specificity, sensitivity, positive predictive value, and negative predictive value were found to be 71.4, 100, 93.1, and 100%, respectively, while these values were 100% for RIF. In addition, the results of the susceptibility test performed with blood agar were obtained on day 14 of incubation. In conclusion, results were obtained much earlier with blood agar (2 weeks) than with 7H10 agar (3 weeks), and the results of this study suggest that blood agar may be used as an alternative medium for the susceptibility testing of M. tuberculosis to INH and RIF. PMID- 15815023 TI - Comparison of the quantitative formalin ethyl acetate concentration technique and agar plate culture for diagnosis of human strongyloidiasis. AB - The quantitative formalin ethyl acetate concentration technique (QFEC) was compared to agar plate culture (APC) for the detection of Strongyloides stercoralis larvae. QFEC could substitute for APC only when the parasite load was higher than 50 larvae per g of stool. This study serves as a good reminder to those conducting stool exams about the sensitivity and specificity limitations of both techniques. PMID- 15815024 TI - Comparison between rpoB and 16S rRNA gene sequencing for molecular identification of 168 clinical isolates of Corynebacterium. AB - Higher proportions (91%) of 168 corynebacterial isolates were positively identified by partial rpoB gene determination than by that based on 16S rRNA gene sequences. This method is thus a simple, molecular-analysis-based method for identification of corynebacteria, but it should be used in conjunction with other tests for definitive identification. PMID- 15815025 TI - Human immunodeficiency virus type 1 in illicit-drug solutions used intravenously retains infectivity. AB - The stability of the human immunodeficiency virus type 1 (HIV-1) strain IIIB in drug solutions was studied. The data demonstrate that HIV-1 infectivity can be retained in drug solutions (e.g. , heroin, "Khanka," and "Vint") for long periods of time. This fact must be taken into account when designing health education programs for the prevention of HIV and AIDS in Eastern Europe. PMID- 15815026 TI - Multiplex PCR for typing strains of Toxoplasma gondii. AB - A multiplex PCR assay was designed for multilocus strain typing of Toxoplasma gondii based on length polymorphism of five microsatellite markers. Eight T. gondii strains already sequenced at these five markers were used as control isolates. This method is simple, rapid, reproducible, and adapted to a large set of isolates. PMID- 15815027 TI - Improvement of the specificity of an enzyme-linked immunosorbent assay for diagnosis of paracoccidioidomycosis. AB - In an attempt to improve the specificity of an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of paracoccidioidomycosis (PCM), sera from patients with PCM were tested using various approaches, such as sodium metaperiodate antigen (gp43) treatment, a serum absorption process with Candida albicans or Histoplasma capsulatum antigens, and dilution of serum in galactose, the main common epitope among pathogenic fungi. The maximum specificity found in this ELISA was 84%. All of these procedures proved inefficient for eliminating all cross-reacting antibodies and obtaining an ELISA specific for PCM diagnosis. PMID- 15815028 TI - Direct identification of major blood culture pathogens, including Pseudomonas aeruginosa and Escherichia coli, by a panel of fluorescence in situ hybridization assays using peptide nucleic acid probes. AB - Rapid identification of four major pathogens from 1,231 positive blood cultures by fluorescence in situ hybridization with peptide nucleic acid probes (AdvanDx Inc., Woburn, Mass.) was evaluated. For Escherichia coli, Staphylococcus aureus, and Candida albicans results agreed with conventional identification. The lower sensitivity of the Pseudomonas aeruginosa assay should not compromise the utility of the four assays. PMID- 15815029 TI - Detection of human metapneumovirus in clinical samples by immunofluorescence staining of shell vial centrifugation cultures prepared from three different cell lines. AB - Monoclonal antibody MAb-8 was evaluated for detection of human metapneumovirus (HMPV) in shell vial centrifugation cultures (SVCC). Detection of HMPV was similar in A549, HEp-2, and LLC-MK2 SVCC, and MAb-8 staining was optimal on day 2 postinoculation. Availability of SVCC for HMPV will be of significant benefit to clinical laboratories. PMID- 15815030 TI - Latex agglutination test for monitoring antibodies to avian influenza virus subtype H5N1. AB - A latex agglutination test (LAT) based on polystyrene beads sensitized with inactivated avian influenza virus H5N1 particles was developed. Compared with the hemagglutination inhibition test, the sensitivity and specificity of the LAT were 88.8 and 97.6%, respectively, in detecting 830 serum samples from vaccinated chickens. The test has application potential in field practice. PMID- 15815031 TI - Evaluation of two chromogenic agar media for recovery and identification of Staphylococcus aureus small-colony variants. AB - To identify the most rapid and reliable technique for recovery and identification of Staphylococcus aureus small-colony variants (SCVs), the colonial appearance of 106 isolates representing SCVs and the normal phenotype were evaluated on two newly described chromogenic agar media. Although almost all of the SCVs grew on the chromogenic agar media, they did not exhibit a change of color. In comparison with conventional media, S. aureus ID agar (SAID; bioMerieux, La Balme Les Grottes, France) showed the most reliable results, with 49 of 53 SCVs tested growing either as an SCV colony or with a normal phenotype after only 24 h of incubation. Growth of SCVs was often not detected before 72 h of incubation on some of the media tested. In conclusion, the most accurate and rapid method to detect both the species S. aureus and the SCV phenotype is to inoculate specimens onto both Columbia blood agar and SAID. PMID- 15815032 TI - Controlled clinical comparison of plastic and glass bottles of BacT/ALERT FA medium for culturing organisms from blood of adult patients. AB - A new, clear-plastic nonvented aerobic FA bottle, designed to prevent breakage, has been developed for the BacT/ALERT blood culture system. We assessed the new plastic FA bottle by comparing its performance with that of the current glass FA bottle for recovery of microorganisms and time to detection of growth in blood samples obtained for culture from adult patients with suspected bloodstream infections. We conclude that the BacT/ALERT plastic and glass FA bottles are comparable for recovery of microorganisms and that the safety advantage of plastic bottles can be achieved without compromising performance. PMID- 15815033 TI - Identity and prevalence of multilocus sequence typing-defined clones of group A streptococci within a hospital setting. AB - Between July and October 2003, 121 clinical isolates of group A streptococci (GAS) were collected from a London hospital and characterized by multilocus sequence typing (MLST) to determine the identity and prevalence of clones circulating within this setting. A total of 39 sequence types (ST), of which 20 were represented by a single isolate, were identified. The eight most prevalent clones among the 121 GAS were ST117/emm81 (16%), ST39/emm4 (9%), ST62/emm87 (7%), ST28/emm1 (6%), ST36/emm12 (6%), ST46/emm22 (5%), ST334/emm82 (5%), and ST101/emm89 (4%). Compared to those in the MLST database (http://spyogenes.mlst.net), 12 (31%) of the 39 STs had not been previously identified, although 7 of these differed from recognized STs at only a single locus, suggesting they were closely related to previously recognized strains. Resistance to erythromycin and tetracycline was seen in 7 and 20% of isolates, respectively, with four isolates resistant to both agents. GAS strains with higher (>80) emm types accounted for 45% of GAS isolates collected during this study. Continuing GAS surveillance, using easily comparable methods, is important for detecting changes in the character of disease-causing isolates. PMID- 15815034 TI - Heterogeneity among strains of diffusely adherent Escherichia coli isolated in Brazil. AB - One hundred twelve diffusely adherent Escherichia coli strains isolated from children in a case control study were evaluated for virulence-associated characteristics, serotyping, antibiotic resistance, and plasmid profiles. Half of the strains hybridized with the probes for icuA (aerobactin) and fimH (type 1 pili); daaE (F1845 fimbriae), afa (afimbrial Dr adhesin), agg-3A (aggregative adhesion fimbria type III fimbriae), pap (P fimbriae), astA (EAST1 toxin), and shET1 (Shigella enterotoxin 1) sequences were present in <20% of the strains. The shET1 gene was noted most frequently in strains isolated from patients. A minority (7%) of the strains produced hemolysin or colicin or showed cytotoxic effects on Vero cells. Forty-five different serotypes were found. The majority (70%) of the strains presented multiple antibiotic resistance. Antibiotic resistance and diffuse adherence were located on the same conjugative plasmids. These results suggest that the transfer of these potential virulence markers could be important in the epidemiology of diffusely adherent E. coli. PMID- 15815035 TI - Evidence for icaADBC-independent biofilm development mechanism in methicillin resistant Staphylococcus aureus clinical isolates. AB - Synthesis of a polysaccharide adhesin by icaADBC-encoded enzymes is currently the best-understood mechanism of staphylococcal biofilm development. In four methicillin-resistant Staphylococcus aureus isolates, environmental activation of icaADBC did not always correlate with increased biofilm production. Moreover, glucose-mediated biofilm development in these isolates was icaADBC independent. Apparently, an environmentally regulated, ica-independent mechanism(s) of biofilm development exists in S. aureus clinical isolates. PMID- 15815036 TI - Evaluation of a rapid assay as an alternative to conventional enzyme immunoassays for detection of hepatitis C virus-specific antibodies. AB - A rapid membrane flow-through immunoassay to detect antibodies to hepatitis C virus was compared with a commercial enzyme immunoassay (EIA) and microparticle enzyme immunoassay (MEIA) using 2,590 serum samples. Sensitivity and specificity of the "rapid assay" in comparison to the EIA/MEIA were 99.3 and 99.0%; the correlation coefficient being 0.91. This assay is suitable where infrastructure and laboratory expertise are limited. PMID- 15815037 TI - Application of a molecular panel to demonstrate enterotropic virus shedding by healthy and human immunodeficiency virus-infected patients. AB - We used a molecular panel, targeting seven enteric viruses, to explore the advantage of using molecular methods to establish the etiology of enteric diseases and to evaluate the prevalence of enteric viruses in asymptomatic human immunodeficiency virus-infected patients. This approach favors rapidity and sensitivity of laboratory diagnosis of viral enteric syndromes. PMID- 15815038 TI - Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus. AB - A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA. PMID- 15815039 TI - First report of infection with community-acquired methicillin-resistant Staphylococcus aureus in South America. AB - Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) has recently emerged in the southwestern Pacific, North America, and Europe. These S. aureus isolates frequently shared some genetic characteristics, including the SCCmec type IV and lukS-lukF genes. In this paper we show that typical CA-MRSA isolates have spread to South America (Brazil). PMID- 15815040 TI - Use of unmodified starches and partial removal of serum to improve Granada medium stability. AB - The use of 1% unmodified rice starch and 1% horse serum instead of 2% soluble starch and 5% serum in Granada medium is described. These components result in a medium of increased stability, preventing spoilage after a few days of storage at room temperature. PMID- 15815041 TI - Characterization of a Bacillus anthracis isolate causing a rare case of fatal anthrax in a 2-year-old boy from Hong Kong. AB - We used multiple-locus variable-number tandem repeat analysis (MLVA) and pagA sequencing to genotype a Bacillus anthracis isolate from a fatal case of human anthrax in Hong Kong. The isolate has a unique MLVA genotype, is related to the Sterne and Ames strains, and is consistent with genotypes identified in China. PMID- 15815042 TI - Use of single nucleotide polymorphisms in the plcR gene for specific identification of Bacillus anthracis. AB - A TaqMan-minor groove binding assay designed around a nonsense mutation in the plcR gene was used to genotype Bacillus anthracis, B. cereus, and B. thuringiensis isolates. The assay differentiated B. anthracis from these genetic near-neighbors and determined that the nonsense mutation is ubiquitous across 89 globally and genetically diverse B. anthracis strains. PMID- 15815043 TI - Fusarium verticillioides abscess of the nasal septum in an immunosuppressed child: case report and identification of the morphologically atypical fungal strain. AB - Morphologically atypical Fusarium verticillioides causing a nasal abscess in a severely immunosuppressed child was successfully treated with repeated surgical intervention and liposomal amphotericin B, despite amphotericin B resistance in vitro. Definitive identification was achieved by sequencing the translation elongation factor alpha gene after ribosomal sequencing proved inadequate. PMID- 15815044 TI - Long-lasting CD3+ T-cell deficiency after cord blood stem cell transplantation in a human herpesvirus 6-infected child. AB - We report a long-lasting (8-month) reactivation of human herpesvirus 6 (HHV-6) infection in child who had undergone cord blood stem cell transplantation. The reactivation was characterized by high viral loads and by immediate-early mRNA positivity. HHV-6 infection was associated with a deep depletion of CD3, while the CD4/CD8 ratio remained substantially unchanged. PMID- 15815046 TI - The polypore mushroom Irpex lacteus, a new causative agent of fungal infections. AB - Irpex lacteus, a wood-decaying basidiomycete, was isolated from a pulmonary abscess of an immunosuppressed child. This medical strain was compared morphologically and by sequencing of the ribosomal intergenic spacers with specimens from both culture collections and herbarium desiccated material. The patient was treated successfully with amphotericin B. PMID- 15815045 TI - Multilocus sequence typing directly on DNA from clinical samples and a cultured isolate to investigate linked fatal pneumococcal disease in residents of a shelter for homeless men. AB - Two apparently linked fatal cases of pneumococcal meningitis were investigated. Pneumolysin PCR performed on blood and cerebrospinal fluid was positive in a culture-negative case. A second case yielded Streptococcus pneumoniae from blood culture. Multilocus sequence typing performed on DNA extracted from case 1 (specimens) and case 2 (isolate) revealed identical sequence types (ST53) substantiating the link between them. PMID- 15815047 TI - Surgery and treatment with high-dose liposomal amphotericin B for eradication of craniofacial zygomycosis in a patient with Hodgkin's disease who had undergone allogeneic hematopoietic stem cell transplantation. AB - This case report describes craniofacial zygomycosis in a 24-year-old male with Hodgkin's disease who underwent chemotherapy and autologous hematopoietic stem cell transplantation, followed by a nonmyeloablative allogeneic transplant. Empirical therapy with itraconazole and amoxicillin-clavulanate failed to resolve the infection. Postdiagnosis, surgery and treatment with high-dose liposomal amphotericin B eradicated the disease. PMID- 15815048 TI - Misidentification of Mycobacterium peregrinum, the causal organism of a case of bacteremia and automatic implantable cardioverter defibrillator-associated infection, due to its unusual acid-fast staining characteristics. AB - We report an unusual case of Mycobacterium peregrinum bacteremia and infection of an automatic implantable cardioverter defibrillator that was originally misidentified as a Nocardia sp. due, in part, to its partially acid-fast staining characteristic, morphology, and odor. The misdiagnosis had a direct effect on patient care, though the patient was subsequently successfully treated. PMID- 15815049 TI - Bacteremia caused by Clostridium intestinale. AB - We describe a case of Clostridium intestinale bacteremia in a previously healthy adolescent female presenting with fever and abdominal pain. The bacterium was definitively identified via 16S rRNA gene sequencing. This is the first report, in the world literature, of human infection caused by this microorganism. PMID- 15815050 TI - Chryseobacterium indologenes non-catheter-related bacteremia in a patient with a solid tumor. AB - A case of non-catheter-related bacteremia caused by Chryseobacterium indologenes in a nonneutropenic man with a solid tumor is described. The patient was successfully treated with piperacillin-tazobactam. PMID- 15815051 TI - Clostridium tetani Is a phospholipase (lecithinase)-producing bacterium. PMID- 15815052 TI - High frequency of mutations in the rpoB gene in rifampin-resistant clinical isolates of Mycobacterium tuberculosis from Singapore. PMID- 15815053 TI - Is the cefoxitin disk test reliable enough to detect oxacillin resistance in coagulase-negative staphylococci? PMID- 15815054 TI - Comparison of two real-time PCR methods for diagnosis of norovirus infection in outbreak and community settings. PMID- 15815055 TI - Reemergence of staphylococcal toxic shock syndrome in the United States since 2000. PMID- 15815056 TI - Successful search-and-destroy policy for methicillin-resistant Staphylococcus aureus in The Netherlands. PMID- 15815058 TI - Transurethral resection of the prostate: reaffirming the gold standard. PMID- 15815057 TI - Cryptic-plasmid-free gonococci may contribute to failure of cppB gene-based assays to confirm results of BD ProbeTEC PCR for identification of Neisseria gonorrhoeae. PMID- 15815059 TI - Surgical intervention for benign prostatic hyperplasia in Hong Kong. AB - OBJECTIVES: To review the treatment of benign prostatic hyperplasia in Hong Kong. DESIGN: Questionnaire study and review of previous presentations at the Hong Kong Urological Association meetings. SETTING: Urology centres in the public sector, Hong Kong. PARTICIPANTS: Thirteen public urology centres replied to the questionnaire survey. Thirty-two papers on benign prostatic hyperplasia presented at past annual scientific meetings of the Hong Kong Urological Association were identified. This would provide an overview of the development of surgical interventions for the treatment of benign prostatic hyperplasia carried out in Hong Kong. RESULTS: Most known surgical techniques have been practised in Hong Kong. Many 'minimally invasive procedures' have now fallen out of favour. Some newer techniques are now available and have produced favourable results. Nonetheless, transurethral resection of the prostate remains the standard surgical intervention in 12 of 13 centres in Hong Kong. CONCLUSION: Transurethral resection of the prostate remains the technique of choice for the treatment of benign prostatic hyperplasia in Hong Kong. New techniques should be comprehensively studied and evaluated before being introduced into regular practice. PMID- 15815060 TI - Causes of childhood blindness in a school for the visually impaired in Hong Kong. AB - OBJECTIVE: To identify the causes of blindness in children attending a school for the blind in Hong Kong. DESIGN: Cross-sectional observational study. SETTING: School for blind children in Hong Kong. PARTICIPANTS: Eighty-two blind students at the Ebenezer School and Home for the Visually Impaired were examined between December 1998 and August 1999. MAIN OUTCOME MEASURES: Demographic data were obtained from students and a questionnaire assessment made of their medical and ocular history. Visual acuity was assessed and visual loss classified according to the World Health Organization classification of visual impairment. Complete ophthalmic assessments were performed in all students including slit-lamp examination and dilated binocular indirect ophthalmoscopy. RESULTS: The mean age of the students was 12.2 years. Ten (12.2%) had a family history of eye disease. Major past medical illnesses were reported in 50% with prematurity and diseases of the central nervous system found in 26.8% and 11.0% of students, respectively. The most common anatomical site for visual impairment was the retina (47.6%), followed by diseases of the optic nerve (14.6%), and diseases of the anterior segment and the lens (14.6%). CONCLUSIONS: The pattern of childhood blindness in Hong Kong is similar to that seen in other developed countries. Preventable causes of childhood blindness, such as prematurity and birth asphyxia, were responsible for a large proportion of cases. Early diagnosis and treatment of such conditions may reduce the incidence of childhood blindness in Hong Kong. PMID- 15815061 TI - Primary immunodeficiency in Hong Kong and the use of genetic analysis for diagnosis. AB - OBJECTIVES: To review the management of primary immunodeficiency and discuss recent advances in genetic analysis. DESIGN: Retrospective study. SETTING: University teaching hospital, Hong Kong. PATIENTS: Children diagnosed with primary immunodeficiency and followed up in the immunology clinic during the period 1988 to 2003. MAIN OUTCOME MEASURES: Demographic data, co-morbidities and treatment of patients, outcome and complications; identification of disease by genetic mutations. RESULTS: Medical records of a total of 117 patients (72 male, 45 female) diagnosed with primary immunodeficiency in the Department of Paediatrics and Adolescent Medicine, Queen Mary Hospital, Hong Kong during the past 15 years (1988-2003) were reviewed. All patients were followed up in the immunology clinic. Some patients had been referred from the private sector or other hospitals for immunological workup. Six categories of primary immunodeficiency were identified: predominantly humoral defect (n=50), predominantly cellular defect (n=22), combined humoral and cellular defect (n=5), phagocytic defect (n=18), complement disorders (n=4), and others (n=18). Although infection was the underlying cause of most co-morbidities and mortality, autoimmune (n=7) and allergic (n=23) manifestations were common. In addition, three patients developed lymphoma. Recent advances in the genetic diagnosis of several types of primary immunodeficiency were also reviewed: X-linked Wiskott Aldrich syndrome, X-linked chronic granulomatous disease, X-linked agammaglobulinaemia, X-linked lymphoproliferative syndrome, leukocyte adhesion disease type I, and X-linked hyperimmunoglobulin M syndrome. This provides an invaluable means of understanding the molecular basis of primary immunodeficiency and has important clinical applications. CONCLUSIONS: Co-morbidities like autoimmune disease and allergic disease are common in patients with primary immunodeficiency and should be carefully evaluated. Likewise, a diagnosis of primary immunodeficiency should be considered when evaluating patients with these conditions. Rapid progress in the field of molecular genetics will enable definite and early diagnosis, and more importantly, potential curative therapy to be administered. PMID- 15815062 TI - Assessment of the Pediatric Index of Mortality (PIM) and the Pediatric Risk of Mortality (PRISM) III score for prediction of mortality in a paediatric intensive care unit in Hong Kong. AB - OBJECTIVE: To compare two models (The Pediatric Risk of Mortality III score and Pediatric Index of Mortality) for prediction of mortality in a paediatric intensive care unit in Hong Kong. DESIGN: Prospective case series. SETTING: A five-bed paediatric intensive care unit in a general hospital in Hong Kong. PATIENTS: All patients consecutively admitted to the unit between April 2001 and March 2003. MAIN OUTCOME MEASURES: Scores for both models compared with observed mortality. RESULTS: A total of 303 patients were admitted to the paediatric intensive care unit during the study period. The median age was 2 years, with an interquartile range of 7 months to 7 years. The male to female ratio was 169:134 (55.8%:44.2%). The median length of hospital stay was 3 days. The overall predicted number of deaths using The Pediatric Risk of Mortality III score was 10.2 patients whereas that by Pediatric Index of Mortality was 13.2 patients. The observed mortality was eight patients. The area under the receiver operating characteristics curve for the two models was 0.910 and 0.912, respectively. CONCLUSION: The predicted mortality using both prediction models correlated well with the observed mortality. PMID- 15815063 TI - Arthroscopic procedures for the treatment of anterior shoulder instability: local experiences. AB - OBJECTIVE: To review the outcomes of arthroscopic stabilisation procedures for the treatment of recurrent anterior shoulder dislocation. DESIGN: Retrospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Patients receiving arthroscopic stabilisation procedures for recurrent anterior shoulder dislocation between 1999 and 2003. MAIN OUTCOME MEASURES: Functional outcomes including pain, range of motion, and activity level were assessed using the Constant score. Intra operative findings were also discussed. RESULTS: A total of 18 arthroscopic stabilisation procedures were performed for the treatment of recurrent shoulder instability. Two cases converted to open procedures were excluded from this review. The overall outcomes were good and seven patients reported a full recovery. Fourteen out of 16 patients reported minimal or no pain, and the mean Constant score was 80. There were no cases of re-dislocation and no major complication was noted. CONCLUSION: All the reviewed patients had a satisfactory functional recovery. Therefore, we believe that the use of arthroscopic stabilisation procedure can produce a favourable outcome for appropriate shoulder pathologies. PMID- 15815064 TI - Unilateral epistaxis after swimming in a stream. AB - A 55-year-old Chinese woman presented with a 3-week history of unilateral left sided epistaxis and nasal obstruction. She had swam in a freshwater stream 1 month prior to the onset of symptoms. Endoscopic examination revealed a live leech at the left middle meatus with a large part of its body inside the left maxillary antrum. Local anaesthetic was applied to anaesthetise the leech and facilitate removal. Magnetic resonance imaging performed following removal confirmed that no other leeches were present in the sinonasal area. The endoparasitism might have persisted because of the inconspicuous site of infestation and the absence of pain. This form of leech infestation has not been previously reported. PMID- 15815065 TI - Cervical meningocele with tethered cervical cord in a Chinese infant. AB - Cervical meningocele and myelomeningocele are rare spinal dysraphic lesions. Unlike lumbosacral dysraphic lesions, there is often no neurological deficit in infants with cervical lesions, thus the subtle features of cervical cord tethering may be overlooked on imaging. We report a case of cervical meningocele in an 8-month-old girl. The tethering band, confirmed intra-operatively, was not evident on imaging. Untethering of the cord was performed together with resection of the sac and repair of the dura. Tethered cord should be suspected in the presence of cervical meningocele and intact neurology. It should be carefully looked for using high-resolution magnetic resonance imaging or computed tomography. Treatment aims to prevent future neurological deterioration, and should include careful intradural exploration with untethering of the cord. PMID- 15815066 TI - Imaging of nail-patella syndrome. AB - Nail-patella syndrome is a rare disorder, which is inherited as an autosomal dominant trait. This condition is also known as hereditary osteo-onychodysplasia or Fong's syndrome. Posterior iliac horns are commonly found in this syndrome and are considered pathognomonic. In this report, we describe the appearance, location, and structure of iliac horns with respect to radiography and magnetic resonance imaging. PMID- 15815067 TI - Candida tropicalis arthritis of the knee in a patient with acute lymphoblastic leukaemia: successful treatment with caspofungin. AB - Candida arthritis in patients with a haematological malignancy is rare. We report a case of Candida tropicalis arthritis of the knee that occurred in a patient with acute lymphoblastic leukaemia during the recovery phase of post-chemotherapy neutropenia. Although the Candida tropicalis isolates from synovial fluid and synovial tissue were sensitive to fluconazole in vitro, a 6-week course of oral treatment failed to produce clinical improvement. The arthritis resolved after 7 weeks of combination therapy with caspofungin, a new echinocandin class of antifungal agent that acts primarily on the cell wall. Eleven other reports of Candida arthritis in patients with a haematological malignancy were reviewed. PMID- 15815068 TI - Resistance to thyroid hormone in a Chinese family with R429Q mutation in the thyroid hormone receptor beta gene. AB - The combination of elevated serum levels of free thyroid hormones with non suppressed thyroid-stimulating hormone suggests the differential diagnoses of resistance to thyroid hormone or thyroid-stimulating hormone-secreting pituitary tumour. Clinical differentiation of these two conditions can be difficult, because patients with thyroid hormone resistance may exhibit various combinations of hypermetabolic and hypometabolic features, and laboratory results have limited sensitivity and specificity. We report a case of resistance to thyroid hormone in a Chinese family that illustrates this difficulty. The diagnosis could only be confirmed by the identification of a known disease-causing mutation in the thyroid hormone receptor beta gene in peripheral leukocytes. Availability of genetic tests will identify more cases in the future and improve our understanding of this condition. PMID- 15815069 TI - Red imported fire ants in Hong Kong. PMID- 15815070 TI - Legal recognition of advance refusal needed. PMID- 15815071 TI - Stapled haemorrhoidectomy in Chinese patients. PMID- 15815072 TI - Vioxx withdrawal--an opportunity to review primary care management for osteoarthritis. PMID- 15815074 TI - Impaired oxidant/antioxidant status and LDL-fatty acid composition are associated with increased susceptibility to peroxidation of LDL in diabetic patients. AB - This study was carried out to determine the relationships between oxidant/antioxidant status, in vitro LDL oxidizability and LDL-fatty acid composition in diabetes mellitus. Plasma total antioxidant capacity (oxygen radical absorbance capacity, ORAC) and LDL-cholesteryl ester fatty acids were investigated in type 1 and type 2 diabetic subjects with and without complications. The degree of LDL oxidation was determined by the measurement of hydroperoxide levels before and after in vitro peroxidative stress with CuSO4. ORAC values were decreased in diabetic subjects who showed high basal hydroperoxide levels. Oxidizability of LDL in these subjects was higher than in control subjects and it was unrelated to LDL-fatty acid composition. However, in type 2 diabetic subjects with complications, alterations in LDL-fatty acid composition were associated with their enhanced oxidative susceptibility. LDL fatty acid alterations might be an additional factor that influences LDL oxidizability especially in type 2 diabetes. In conclusion, diabetes mellitus is associated with enhanced oxidative stress and defective antioxidant/oxidant balance regardless the type of diabetes and presence of complications. PMID- 15815075 TI - Time course of ischemia/reperfusion-induced oxidative modification of neural proteins in rat forebrain. AB - Time course of oxidative modification of forebrain neural proteins was investigated in the rat model of global and partial cerebral ischemia/reperfusion. Animals were subjected to 4-vessel occlusion for 15 min (global ischemia). After the end of ischemia and at different reperfusion times (2, 24 and 48 h), lipoperoxidation-dependent and direct oxidative modification neural protein markers were measured in the forebrain total membrane fraction (tissue homogenate). Ischemia itself causes significant changes only in levels of tryptophan and bityrosine fluorescence when compared to controls. All tested parameters of protein modification altered significantly and were maximal at later reperfusion stage. Content of carbonyl group in re-flow period steadily increased and culminated at 48 h of reperfusion. The highest increase in the fluorescence of bityrosines was detected after 24 h of reperfusion and was statistically significant to both sham operated and ischemic groups. The changes in fluorescence intensity of tryptophan decreased during a reperfusion time dependent manner. Formation of lysine conjugates with lipoperoxidation end products significantly increased only at later stages of reperfusion. Total forebrain membranes from animals subjected to 3-vessel occlusion model to 15 min (partial ischemia) show no altered content of oxidatively modified proteins compared to controls. Restoration of blood flow for 24 h significantly decreased only fluorescence of aromatic tryptophan. Partial forebrain ischemia/reperfusion resulted in no detectable significant changes in oxidative products formation in extracerebral tissues (liver and kidney) homogenates. Our results suggest that global ischemia/reperfusion initiates both the lipoperoxidation-dependent and direct oxidative modifications of neural proteins. The findings support the view that spatial and temporal injury at later stages of ischemic insult at least partially involves oxidative stress-induced amino acid modification. The results might have important implications for the prospective post-ischemic antioxidant therapy. PMID- 15815076 TI - Retinoic acid increased expression of the Na+/Ca2+ exchanger in the heart and brain. AB - The Na+/Ca2+ exchanger (NCX) is an important calcium transport system, which regulates intracellular calcium homeostasis. In particular, NCX is highly expressed in the plasma membrane of excitable neuronal and cardiac cells. We report here that binding of retinoic acid enhances expression of the NCX1 in the rat cardiac atria, brain stem and hypothalamus, but not in cerebellum. Differences in the regulation of the NCX1 by retinoic acid might suggest that GATA4-retinoic acid inducible transcription factor is activated in the hypothalamus and brain stem, but not in the cerebellum. Our results support the idea that inducible transcription factors play an important role in the fine tuning of local tissue calcium homeostasis. PMID- 15815077 TI - Rabbit liver microsomal system: study of interaction with two model N nitrosamines and their metabolism. AB - Rabbit liver microsomes of control (non-treated) or animals induced either by ethanol (EtOH) or phenobarbital (PB) were incubated with N-nitrosodimethylamine (NDMA) or N-nitrosomethylaniline (NMA). Difference spectroscopy showed that NMA is bound to the substrate-binding site of cytochrome P-450 (CYP) isoforms as heme ligand in control and EtOH pre-treated microsomes. On the other hand, PB-induced microsomes exhibit with NMA substrate type of spectra. NDMA does not provide any type of binding spectra with used microsomal systems. Oxidative bio-activation of N-nitrosamines by the microsomal CYP isoforms was measured as formaldehyde formation. Analysis of reaction kinetics in control microsomes revealed, for both substrates, two values of Michaelis-Menten constant (K(m)) for, K(m) values of 0.03 and 0.13 mmol/l for NDMA, and 0.30 and 0.82 mmol/l for NMA. Induction of animals with EtOH resulted in a decrease in the K(m) value for both substrates. In contrast, PB treatment caused an elevation of K(m) value for NDMA. Based on these data, we conclude that EtOH-inducible microsomal CYP isoforms (mainly CYP2E1) are responsible for binding and N-demethylation metabolism of both studied N-nitrosamines in rabbit liver microsomal system. The role of the other CYP isoforms involved in the metabolism of mentioned N-nitrosamines is discussed. PMID- 15815078 TI - Factor V Leiden in patients with venous thrombosis in Slovak population. AB - Resistance to activated protein C determined by factor V Leiden (FVL) is the most frequent inherited risk factor of venous thrombosis. The purpose of our work was to reveal the frequency of FVL in Slovak patients with venous thromboses, to characterise the nature of venous thromboses in this inherited thrombophilia, and to consider the screening approach to investigation of FVL in patients with venous thromboses. 350 patients with a diagnosis of venous thromboembolic disease from various regions of Slovakia were investigated. FVL, detected by polymerase chain reaction, was found in 128/350 (37%) patients with venous thromboses. 118/128 (92%) patients were heterozygous and 10/128 (8%) were homozygous carriers. In 108/128 (84%) patients with FVL the thromboembolic disease occurred spontaneously. Phlebothrombosis occurred predominantly in the lower limbs- 117/128 (91%) patients, atypical localisations were rare. The first thromboembolic event was manifested before 40 years of age in 69% of patients. The family history was positive in 60/128 (47%) FVL carriers with thromboembolic disease. Recurrent thrombosis occurred in 30% of patients with FVL. In agreement with findings in other European countries, the prevalence of FVL was high in Slovak patients with thromboembolic disease. The investigation of FVL seems to be justified in patients before 40 years of age with venous thrombosis of lower limbs, in the absence of triggering factors and with a family history of venous thromboembolic disease. PMID- 15815079 TI - Amiloride-sensitive sodium transport in lamprey red blood cells: evidence for two distinct transport pathways. AB - To determine Na+/H+ exchange in lamprey erythrocyte membranes, the cells were acidified to pH(i) 6.0 using the K+/H+ ionophore nigericin. Incubation of acidified erythrocytes in a NaCl medium at pH 8.0 caused a considerable rise in 22Na+ influx and H+ efflux during the first 1 min of exposure. In addition, exposure of acidified red cells to NaCl medium was associated with rapid elevation of intracellular Na+ content. The acid-induced changes in Na+ influx and H+ efflux were almost completely inhibited by amiloride and dimethylamiloride. In native lamprey erythrocytes, amiloride-sensitive Na+ influx progressively increased as the osmolality of incubation medium was increased by addition of 100, 200, or 300 mmol/l sucrose. Unexpectedly, the hypertonic stress induced a small, yet statistically significant decrease in intracellular Na+ content in these cells. The reduction in the cellular Na+ content increased with hypertonicity of the medium. The acid- and shrinkage-induced Na+ influxes were inhibited by both amiloride and 5-(N-ethyl-N-isopropyl)-amiloride (EIPA) in a dose-dependent manner. For both blockers, the half-maximal inhibitory values (IC50) were much greater for the shrinkage-induced (44 and 15 micromol/l for amiloride and EIPA, respectively) than for the acid-induced Na+ influx (5.1 and 3.3 micromol/l, respectively). The data obtained are the first demonstration of the presence of a Na+/H+ exchanger with high activity in acidified (pH(i) 6.0) lamprey red blood cells (on average, 512 +/- 56 mmol/l cells/h, n = 13). The amiloride-sensitive Na+ influxes produced by hypertonic cell shrinkage and acid load are likely to be mediated by distinct ion transporters in these cells. PMID- 15815080 TI - Effect of Tanakan on postischemic activity of protein synthesis machinery in the rat brain. AB - A growing body of evidence supports the role of free radicals in triggering the functional and metabolic disturbances following transient cerebral ischemia. This study was designed to evaluate whether the extent of reperfusion-induced inhibition of protein synthesis initiation as well as tissue injury can be reduced by Tanakan (Ginkgo biloba extract, EGb 761) (Beaufour-Ipsen Industrie). Rats received Tanakan in the dose of 40 mg/kg/day for 7 days before surgical intervention. Transient forebrain ischemia was induced by 4-vessel occlusion. Rats were subjected to 20 min of ischemia followed by 30 min, 4 h or 7 days of reperfusion. Protein synthesis rate, reinitiation ability and neurodegeneration in the frontal cortex and hippocampus were measured by the incorporation of radioactively labelled leucine into polypeptide chains in postmitochondrial supernatants and by Fluoro-Jade B staining. The protective effect was observed, concerning both the protein synthesis and the number of surviving neurons, in the Tanakan-treated groups. Tanakan significantly reduced the ischemia/reperfusion induced inhibition of translation in the neocortex as well as in the highly sensitive hippocampus. Our results indicate that free radicals play an important role in the development of reperfusion-induced injury, and the treatment of ischemic and reperfused brain with free radical scavengers may reduce the severity of reperfusion damage. PMID- 15815081 TI - Evaluation of comparative cytochrome P450 2B4 model by photoaffinity labeling. AB - A homology model of rabbit CYP 2B4 was constructed on the basis of the crystallographic structure of truncated mammalian CYP 2C5/3 and bacterial soluble CYPs. To validate the CYP 2B4 homology model photoaffinity labeling was employed. Three probes (I-III) containing a photo-labile azido-group and an amino-group on opposite ends of the molecule were designed for photoaffinity labeling of the CYP 2B4 in increasing distance from the heme iron. Spectroscopic data proved probes I (the shortest) and II (a middle sized) to be coordinated with the heme iron via their amino-groups in the enzyme active center while the probe III (the longest) was not bound in this way. This binding orientation of probes I and II is in accordance with the model predicting ion-pairing of the negatively charged side chain of CYP 2B4 Asp 105 and a positively charged nitrogen located in an appropriate position in structures of probes I and II, only. The lack of heme binding of the probe III is clear from its docking into the CYP 2B4 model since no Asp 105 ion-pairing is possible. The target of photoactivated probe II, Arg 197, in a distance of about 16.5 A from the heme iron, exactly matches the position of that amino acid residue, predicted from the CYP 2B4 homology model. Moreover, using this technique, a substrate access channel has been identified. To assess the predicted substrate-binding pocket, an interaction of a specific CYP 2B4 substrate, diamantane, was examined. In "silico" docking revealed strong binding of diamantane in an orientation allowing experimentally observed C4 hydroxylation. Our homology model of CYP 2B4 is thus consistent with experimental metabolic and photoaffinity labeling data. PMID- 15815082 TI - Proteins released from liver after ischaemia induced an elevation of heart resistance against ischaemia-reperfusion injury: 2. Beneficial effect of liver ischaemia in situ. AB - We have shown earlier that proteins released from the heart during preconditioning may protect non-preconditioned heart during sustained ischaemia, similarly as preconditioning itself. In other our experiments we have documented that also proteins released from isolated rat liver during reperfusion after global ischaemia performed a protective effect on isolated rat heart against ischaemia-reperfusion injury. In the current study we examined the effect of liver ischaemia in situ on resistance of rat heart to ischaemia and reperfusion injury. Wistar rats (male) were subjected to liver ischaemia maintained by occlusion of portal vein and hepatic artery for 20 min, followed with 30-min reperfusion after reopening of both vessels. Then the hearts were isolated and perfused according to Langendorf. Hearts, after initial stabilisation (15 min), were subjected to 20-min ischaemia and 30-min reperfusion. During reperfusion, the haemodynamic parameters of hearts were measured. The protein pattern of high soluble fraction (HS fraction) isolated from rat blood by precipitation with ammonium sulphate was detected by SDS-PAGE. Our results showed improved parameters of pressure and contractility in the group after liver ischaemia (ischaemic group), presented by decreased diastolic pressure and increased LVDP((S-D)) in comparison with levels of these parameters in the control group. We also observed improved heart contraction-relaxation cycles parameters (dP/dt)(max) and (dP/dt)(min) in ischaemic group as compared with the control group. On the other hand, there were no significant differences in heart rate and coronary flow between both experimental groups. SDS-PAGE showed changed protein pattern in HS fraction, particularly the levels of several low molecular weight proteins increased. We conclude that liver ischaemia induced a higher resistance of heart against ischaemia-reperfusion injury. We propose that release of some cardioprotective proteins present in HS fraction can also contribute to this cardioprotection. PMID- 15815083 TI - Expression of C/EBP delta in rat liver during development and the acute-phase response. AB - Using Western analysis, C/EBP delta was established in the nuclear extract and nuclear matrix throughout rat liver development and in the adult. During the acute-phase response (APR), C/EBP delta increased in the nuclear extract but remained unchanged in the nuclear matrix of fetal and postnatal rats, whereas it increased in both the nuclear extract and nuclear matrix of the adult. The solubility partitioning of gene regulatory proteins in the nucleus is important for their functioning (Uskokovic et al. 2002). The obtained different solubility partitioning profiles of C/EBP delta suggest that its activity is regulated by different mechanisms during development and in the adult. PMID- 15815084 TI - Melatonin scavenges phenylglyoxylic ketyl radicals. AB - The antioxidant properties of melatonin were tested in this work by EPR technique. It was found that melatonin scavenges phenylglyoxylic ketyl radicals. Its effectiveness was 10-times lower than that of vitamin C. A new method of generation of phenylglyoxylic ketyl radicals by spontaneous decomposition of D,L 2,3-diphenyltartaric acid in propan-2-ol was used. PMID- 15815085 TI - Practical and dosimetric implications of a new type of packaging for radiographic film. AB - Recently, Kodak introduced new light-tight packages (vacuum packaging, aluminium layer under black polyethylene and different paper) for their oncology films (EDR 2, X-Omat V and PPL-2). In order to avoid additional uncertainty and to ensure transferability of previously published results, we assessed in this study the effect of the old and new packages on the dosimetric response of EDR-2 radiographic film. Therefore, sensitometric measurements were performed for different film assemblies (new envelope + new paper, old envelope + old paper, new envelope without paper and old envelope without paper). In addition, to assess possible effects of the package on the film depth-dose response, packaged films were irradiated in parallel geometry, and central depth-dose curves were retrieved. For the perpendicular geometry, on the other hand, the effect of the package was assessed at large depth for a high intensity-modulated inverse pyramid beam. The results of the sensitometric measurements reveal no difference between the packages. However, the white colour of the paper in both the packages induces a dose-dependent increase in optical density (0-0.12) of the film. The depth-dose curves show better reproducibility for the new package and the new paper improves the accuracy of film dosimetry, but despite the company's effort to evacuate the air out of the new envelope, it remains necessary to clamp the films in the phantom for the parallel irradiation geometry. At 5 cm depth, the films irradiated in parallel geometry show an under-response of 3-5% compared to films irradiated perpendicularly. Finally, even at locations of large photon scatter, no filtration effect from the aluminium layer incorporated in the new envelope has been observed for perpendicular irradiation geometry. PMID- 15815086 TI - Prediction of density and mechanical properties of human trabecular bone in vitro by using ultrasound transmission and backscattering measurements at 0.2-6.7 MHz frequency range. AB - The ultrasound (US) backscattering method has been introduced as an alternative for the through-transmission measurement of sound attenuation and speed in diagnosis of osteoporosis. Both attenuation and backscattering depend strongly on the US frequency. In this study, 20 human trabecular bone samples were measured in transmission and pulse-echo geometry in vitro. The aim of the study was to find the most sensitive frequency range for the quantitative ultrasound (QUS) analyses. Normalized broadband US attenuation (nBUA), speed of sound (SOS), broadband US backscatter (BUB) and integrated reflection coefficient (IRC) were determined for each sample. The samples were spatially scanned with five pairs of US transducers covering a frequency range of 0.2-6.7 MHz. Furthermore, mechanical properties and density of the same samples were determined. At all frequencies, SOS, BUB and IRC showed statistically significant linear correlations with the mechanical properties or density of human trabecular bone (0.51 < r < 0.82, 0.54 < r < 0.81 and 0.70 < r < 0.85, respectively). In contrast to SOS, IRC and BUB, nBUA showed statistically significant correlations with mechanical parameters or density at the centre frequency of 1 MHz only. Our results suggest that frequencies up to 5 MHz can be useful in QUS analyses for the prediction of bone mechanical properties and density. Since the use of higher frequencies provides better axial and spatial resolution, improved structural analyses may be possible. While extensive attenuation of high frequencies in trabecular bone limits the clinically feasible frequency range, selection of optimal frequency range for in vivo QUS application should be carefully considered. PMID- 15815087 TI - Minimum data image reconstruction algorithms with shift-invariant filtering for helical, cone-beam CT. AB - We derive accurate and efficient reconstruction algorithms for helical, cone-beam CT that employ shift-invariant filtering. Specifically, a new backprojection filtration algorithm is developed, and a minimum data filtered-backprojection algorithm is derived. These reconstruction algorithms with shift-invariant filtering can accept data with transverse truncation, and hence allow for minimum data image reconstruction. PMID- 15815088 TI - Analysis of discrete and continuous distributions of ventilatory time constants from dynamic computed tomography. AB - In this study, an algorithm was developed to measure the distribution of pulmonary time constants (TCs) from dynamic computed tomography (CT) data sets during a sudden airway pressure step up. Simulations with synthetic data were performed to test the methodology as well as the influence of experimental noise. Furthermore the algorithm was applied to in vivo data. In five pigs sudden changes in airway pressure were imposed during dynamic CT acquisition in healthy lungs and in a saline lavage ARDS model. The fractional gas content in the imaged slice (FGC) was calculated by density measurements for each CT image. Temporal variations of the FGC were analysed assuming a model with a continuous distribution of exponentially decaying time constants. The simulations proved the feasibility of the method. The influence of experimental noise could be well evaluated. Analysis of the in vivo data showed that in healthy lungs ventilation processes can be more likely characterized by discrete TCs whereas in ARDS lungs continuous distributions of TCs are observed. The temporal behaviour of lung inflation and deflation can be characterized objectively using the described new methodology. This study indicates that continuous distributions of TCs reflect lung ventilation mechanics more accurately compared to discrete TCs. PMID- 15815090 TI - On the surviving fraction in irradiated multicellular tumour spheroids: calculation of overall radiosensitivity parameters, influence of hypoxia and volume effects. AB - We model the heterogeneous response to radiation of multicellular tumour spheroids assuming position- and volume-dependent radiosensitivity. We propose a method to calculate the overall radiosensitivity parameters to obtain the surviving fraction of tumours. A mathematical model of a spherical tumour with a hypoxic core and a viable rim which is a caricature of a real tumour is constructed. The model is embedded in a two-compartment linear-quadratic (LQ) model, assuming a mixed bivariated Gaussian distribution to attain the radiosensitivity parameters. Ergodicity, i.e., the equivalence between ensemble and volumetric averages is used to obtain the overall radiosensitivities for the two compartments. We obtain expressions for the overall radiosensitivity parameters resulting from the use of both a linear and a nonlinear dependence of the local radiosensitivity with position. The model's results are compared with experimental data of surviving fraction (SF) for multicellular spheroids of different sizes. We make one fit using only the smallest spheroid data and we are able to predict the SF for the larger spheroids. These predictions are acceptable particularly using bounded sensitivities. We conclude with the importance of taking into account the contribution of clonogenic hypoxic cells to radiosensitivity and with the convenience of using bounded local sensitivities to predict overall radiosensitivity parameters. PMID- 15815089 TI - Automated melanoma detection with a novel multispectral imaging system: results of a prospective study. AB - The aim of this research was to evaluate the performance of a new spectroscopic system in the diagnosis of melanoma. This study involves a consecutive series of 1278 patients with 1391 cutaneous pigmented lesions including 184 melanomas. In an attempt to approach the 'real world' of lesion population, a further set of 1022 not excised clinically reassuring lesions was also considered for analysis. Each lesion was imaged in vivo by a multispectral imaging system. The system operates at wavelengths between 483 and 950 nm by acquiring 15 images at equally spaced wavelength intervals. From the images, different lesion descriptors were extracted related to the colour distribution and morphology of the lesions. Data reduction techniques were applied before setting up a neural network classifier designed to perform automated diagnosis. The data set was randomly divided into three sets: train (696 lesions, including 90 melanomas) and verify (348 lesions, including 53 melanomas) for the instruction of a proper neural network, and an independent test set (347 lesions, including 41 melanomas). The neural network was able to discriminate between melanomas and non-melanoma lesions with a sensitivity of 80.4% and a specificity of 75.6% in the 1391 histologized cases data set. No major variations were found in classification scores when train, verify and test subsets were separately evaluated. Following receiver operating characteristic (ROC) analysis, the resulting area under the curve was 0.85. No significant differences were found among areas under train, verify and test set curves, supporting the good network ability to generalize for new cases. In addition, specificity and area under ROC curve increased up to 90% and 0.90, respectively, when the additional set of 1022 lesions without histology was added to the test set. Our data show that performance of an automated system is greatly population dependent, suggesting caution in the comparison with results reported in the literature. In our opinion, scientific reports should provide, at least, the median values of thickness and dimension of melanomas, as well as the number of small (6 mm) melanomas. PMID- 15815091 TI - Development and operation of a pixel segmented liquid-filled linear array for radiotherapy quality assurance. AB - A liquid isooctane (C(8)H(18)) filled ionization linear array for radiotherapy quality assurance has been designed, built and tested. The detector consists of 128 pixels, each of them with an area of 1.7 mm x 1.7 mm and a gap of 0.5 mm. The small pixel size makes the detector ideal for high gradient beam profiles such as those present in intensity modulated radiation therapy (IMRT) and radiosurgery. As the read-out electronics we use the X-ray Data Acquisition System with the Xchip developed by the CCLRC. Studies concerning the collection efficiency dependence on the polarization voltage and on the dose rate have been made in order to optimize the device operation. In the first tests, we have studied dose rate and energy dependences. Dose rate dependence was found to be lower than 2.1% up to 5 Gy min(-1), and energy dependence lower than 2.5% up to 20 cm depth in solid water. Output factors and penumbras for several rectangular fields have been measured with the linear array and were compared with the results obtained with a 0.125 cm(3) air ionization chamber and radiographic film, respectively. Finally, we have acquired profiles for an IMRT field and for a virtual wedge. These profiles have also been compared with radiographic film measurements. All the comparisons show a good correspondence. The device has proved its capability to verify on-line therapy beams with good spatial resolution and signal-to-noise ratio. PMID- 15815092 TI - Evaluation of the dosimetric performance of BANG3 polymer gel. AB - New radiotherapy techniques call for three-dimensional dosimetric methods with high spatial resolution. Radiation sensitive gels read out using MRI T(2) mapping provide an extremely promising option, and commercially available BANG polymer gels provide a convenient route into gel dosimetry. Gel dosimetry is dependent on the ability to calibrate gel response against radiation dose. This in turn is dependent on the reproducibility of response both between gels irradiated to the same dose and for a single gel sample over time. This study aims to evaluate the performance of a commercially available BANG gel. Our experimental arrangement gave excellent precision of radiation delivery (<0.2%) and reproducibility of T(2) measurement (<0.5%). Seven groups of 10 test tubes containing BANG3 gel were irradiated in 0.5 Gy steps between 0 and 3 Gy. A further four groups of four samples were irradiated in 2 Gy steps between 4 and 10 Gy. The gel samples were identical and derived from the same manufacturing batch. MR imaging was carried out four days after irradiation and then at weekly intervals for four weeks. Short-term variation in gel response can readily be corrected using reference samples. Longer term systematic drift of the gel calibration curve was observed relative to reference samples prepared in-house for quality assurance purposes. This implies that read-out of the calibration gels and dosimetry phantom must be performed at the same time after irradiation, or errors of up to 25% may be incurred. Precision of gel response did not change significantly over time. The observation of significantly different T(2) values both prior to irradiation and following irradiation to the same dose (variation up to 15%) illustrates the current difficulties associated with BANG3 gel calibration and constrains the practical utility of these commercially available gels for clinical radiation dosimetry. PMID- 15815093 TI - Technical considerations for implementation of x-ray CT polymer gel dosimetry. AB - Gel dosimetry is the most promising 3D dosimetry technique in current radiation therapy practice. X-ray CT has been shown to be a feasible method of reading out polymer gel dosimeters and, with the high accessibility of CT scanners to cancer hospitals, presents an exciting possibility for clinical implementation of gel dosimetry. In this study we report on technical considerations for implementation of x-ray CT polymer gel dosimetry. Specifically phantom design, CT imaging methods, imaging time requirements and gel dose response are investigated. Where possible, recommendations are made for optimizing parameters to enhance system performance. The dose resolution achievable with an optimized system is calculated given voxel size and imaging time constraints. Results are compared with MRI and optical CT polymer gel dosimetry results available in the literature. PMID- 15815094 TI - A finite size pencil beam for IMRT dose optimization. AB - Dose optimization for intensity modulated radiotherapy (IMRT) using small field elements (beamlets) requires the computation of a large number of very small, often only virtual fields of typically a few mm to 1 cm in size. The primary requirements for a suitable dose computation algorithm are (1) speed and (2) proper consideration of the penumbra of the fields which are composed of these beamlets. Here, a finite size pencil beam (fsPB) algorithm is proposed which was specifically designed for the purpose of beamlet-based IMRT. The algorithm employs an analytical function for the cross-profiles of the beamlets which is based on the assumption of self-consistency, i.e. the requirement that an arbitrary superposition of abutting beamlets should add up to a homogeneous field. The depth dependence is stored in tables derived from Monte Carlo computed dose distributions. It is demonstrated that the algorithm produces accurately the output factors and cross-profiles of typical multi-leaf-shaped segments. Due to the accurate penumbra model, the dose distribution features physically feasible gradients at any stage of the iterative optimization, which eliminates the problem of large discrepancies in normal tissue dose due to misaligned gradients between optimized and recomputed treatment plans. PMID- 15815095 TI - A 3D photon superposition/convolution algorithm and its foundation on results of Monte Carlo calculations. AB - Based on previous publications on a triple Gaussian analytical pencil beam model and on Monte Carlo calculations using Monte Carlo codes GEANT-Fluka, versions 95, 98, 2002, and BEAMnrc/EGSnrc, a three-dimensional (3D) superposition/convolution algorithm for photon beams (6 MV, 18 MV) is presented. Tissue heterogeneity is taken into account by electron density information of CT images. A clinical beam consists of a superposition of divergent pencil beams. A slab-geometry was used as a phantom model to test computed results by measurements. An essential result is the existence of further dose build-up and build-down effects in the domain of density discontinuities. These effects have increasing magnitude for field sizes < or =5.5 cm(2) and densities < or = 0.25 g cm(-3), in particular with regard to field sizes considered in stereotaxy. They could be confirmed by measurements (mean standard deviation 2%). A practical impact is the dose distribution at transitions from bone to soft tissue, lung or cavities. PMID- 15815096 TI - Fast modelling of the collimator-detector response in Monte Carlo simulation of SPECT imaging using the angular response function. AB - Interactions of incident photons with the collimator and detector, including septal penetration, scatter and x-ray fluorescence, are significant sources of image degradation in applications of SPECT including dual isotope imaging and imaging using radioisotopes that emit high- or medium-energy photons. Modelling these interactions using full Monte Carlo (MC) simulations is computationally very demanding. We present a new method based on the use of angular response functions (ARFs). The ARF is a function of the incident photon's direction and energy and represents the probability that a photon will either interact with or pass through the collimator, and be detected at the intersection of the photon's direction vector and the detection plane in an energy window of interest. The ARFs were pre-computed using full MC simulations of point sources that include propagation through the collimator-detector system. We have implemented the ARF method for use in conjunction with the SimSET/PHG MC code to provide fast modelling of both interactions in the patient and in the collimator-detector system. Validation results in the three cases studied show that there was good agreement between the projections generated using the ARF method and those from previously validated full MC simulations, but with hundred to thousand fold reductions in simulation time. PMID- 15815097 TI - Exact fan-beam image reconstruction algorithm for truncated projection data acquired from an asymmetric half-size detector. AB - In this paper, we present a new algorithm designed for a specific data truncation problem in fan-beam CT. We consider a scanning configuration in which the fan beam projection data are acquired from an asymmetrically positioned half-sized detector. Namely, the asymmetric detector only covers one half of the scanning field of view. Thus, the acquired fan-beam projection data are truncated at every view angle. If an explicit data rebinning process is not invoked, this data acquisition configuration will reek havoc on many known fan-beam image reconstruction schemes including the standard filtered backprojection (FBP) algorithm and the super-short-scan FBP reconstruction algorithms. However, we demonstrate that a recently developed fan-beam image reconstruction algorithm which reconstructs an image via filtering a backprojection image of differentiated projection data (FBPD) survives the above fan-beam data truncation problem. Namely, we may exactly reconstruct the whole image object using the truncated data acquired in a full scan mode (2pi angular range). We may also exactly reconstruct a small region of interest (ROI) using the truncated projection data acquired in a short-scan mode (less than 2pi angular range). The most important characteristic of the proposed reconstruction scheme is that an explicit data rebinning process is not introduced. Numerical simulations were conducted to validate the new reconstruction algorithm. PMID- 15815098 TI - A numerical study of transcranial focused ultrasound beam propagation at low frequency. AB - The feasibility of transcranial ultrasound focusing with a non-moving phased array and without skull-specific aberration correction was investigated using computer simulations. Three cadaver skull CT image data sets were incorporated into an acoustic wave transmission model to simulate transskull ultrasound wave propagation. Using a 0.25 MHz hemispherical array (125 mm radius of curvature, 250 mm diameter, 24 255 elements), the simulated beams could be focused and steered with transducer element driving phases and amplitude adjusted for focal beam steering in water (water-path). A total of 82 foci, spanning wide ranges of distance in the three orthogonal dimensions, were simulated to test the focal beam steering capability inside the three skulls. The acoustic pressure distribution in a volume of 20 x 20 x 20 mm(3) centred at each focus was calculated with a 0.5 mm spacing in each axis. Clearly defined foci were retained through the skulls (skull-path) in most cases. The skull-path foci were on average 1.6 +/- 0.8 mm shifted from their intended locations. The -3 dB skull path beam width and length were on average 4.3 +/- 1.0 mm and 7.7 +/- 1.8 mm, respectively. The skull-path sidelobe levels ranged from 25% to 55% of the peak pressure values. The skull-path peak pressure levels were about 10%-40% of their water-path counterparts. Focusing low-frequency beam through skull without skull specific aberration correction is possible. This method may be useful for applying ultrasound to disrupt the blood-brain barrier for targeted delivery of therapeutic or diagnostic agents, or to induce microbubbles, or for other uses of ultrasound in brain where the required power levels are low and the sharp focusing is not needed. PMID- 15815099 TI - Attenuation correction for small animal PET tomographs. AB - Attenuation correction is one of the important corrections required for quantitative positron emission tomography (PET). This work will compare the quantitative accuracy of attenuation correction using a simple global scale factor with traditional transmission-based methods acquired either with a small animal PET or a small animal x-ray computed tomography (CT) scanner. Two phantoms (one mouse-sized and one rat-sized) and two animal subjects (one mouse and one rat) were scanned in CTI Concorde Microsystem's microPET Focus for emission and transmission data and in ImTek's MicroCAT II for transmission data. PET emission image values were calibrated against a scintillation well counter. Results indicate that the scale factor method of attenuation correction places the average measured activity concentration about the expected value, without correcting for the cupping artefact from attenuation. Noise analysis in the phantom studies with the PET-based method shows that noise in the transmission data increases the noise in the corrected emission data. The CT-based method was accurate and delivered low-noise images suitable for both PET data correction and PET tracer localization. PMID- 15815100 TI - Are phantoms useful for predicting the potential of dose reduction in full-field digital mammography? AB - A phantom study was performed in full-field digital mammography to investigate the opportunity and the magnitude of a possible dose reduction that would leave the image quality above the accepted thresholds associated with some classical phantoms. This preliminary work is intended to lay the groundwork for a future clinical study on the impact of dose reduction on clinical results. Three different mammography phantoms (ACR RMI 156, CIRS 11A and CDMAM 3.4) were imaged by a full-field digital mammography unit (GE Senographe 2000D) at different dose levels. Images were rated by three observers with softcopy reading and scoring methods specific to each phantom. Different types of data analysis were applied to the ACR (American College of Radiology) and the other two phantoms, respectively. With reference to the minimum acceptance score in screen/film accreditation programmes, the ACR phantom showed that about 45% dose reduction could be applied, while keeping the phantom scores above that threshold. A relative comparison was done for CIRS and CDMAM, for which no threshold is defined. CIRS scoring remained close to the reference level down to 40% dose reduction, the inter- and intra-observer variability being the main source of uncertainty. Contrast-detail curves provided by CDMAM overlapped down to 50% dose reduction, at least for object contrast values ranging between 30% and 3%. This multi-phantom study shows the potential of further reducing the dose in full field digital mammography beyond the current values. A common dose reduction factor around 50% seems acceptable for all phantoms. However, caution is required before extrapolating the results for clinical use, given the limitations of these widely used phantoms, mainly related to their limited dynamic range and uniform background. PMID- 15815101 TI - The effect of free-electron laser pulse structure on mid-infrared soft-tissue ablation: ablation metrics. AB - Pulsed mid-infrared (6.45 microm) radiation has been shown to cut soft tissue with minimal collateral damage (<40 microm); however, the mechanism of ablation has not been elucidated to date. The goal of this research was to examine the role of the unique pulse structure of the Vanderbilt Mark-III free-electron laser (FEL) and its role in the efficient ablation of soft tissue with minimal collateral damage. The effect of the picosecond micropulse was examined by running the native FEL pulse structure through a pulse stretcher in order to increase the micropulse length from 1 ps up to approximately 200 ps. This allowed us to determine whether or not the picosecond train of micropulses played any role in the ablation process. The ablation threshold was determined for water and mouse dermis for each micropulse length. While the results of the analysis showed a statistically significant difference between 1 and 200 ps, the average per cent difference amounts to only 28% and is not proportional to the 200-fold drop in peak irradiance. The ablation efficiency was also measured on gelatin and mouse dermis for the different micropulse lengths. A small but statistically significant difference was observed between 1 and 200 ps, with the 200 ps pulse being more efficient on gelatin, and with the opposite trend for mouse dermis. We have shown that there is a small effect of micropulse duration of the FEL on the ablation process; however, this effect is negligible between 1 and 200 ps given that there is a 200-fold decrease in peak intensity. These results suggest that as we move forward in developing alternative laser sources for tissue ablation to replace the FEL, the picosecond micropulse structure is not a critical parameter that needs to be duplicated. PMID- 15815102 TI - The effect of free-electron laser pulse structure on mid-infrared soft-tissue ablation: biological effects. AB - Previous studies have shown that changing the pulse structure of the free electron laser (FEL) from 1 to 200 ps and thus reducing the peak irradiance of the micropulse by 200 times had little or no effect on both the ablation threshold radiant exposure and the ablated crater depth for a defined radiant exposure. This study focuses on the ablation mechanism at 6.1 and 6.45 microm with an emphasis on the role of the FEL pulse structure. Three different experiments were performed to gain insight into this mechanism. The first was an analysis of the ablation plume dynamics observed for a 1 ps micropulse compared with a 200 ps micropulse as seen through bright-field analysis. Negligible differences are seen in the size, but not the dynamics of ablation, as a result of this imaging. The second experiment was a histological analysis of corneal and dermal tissue to determine whether there is less thermal damage associated with one micropulse duration versus another. No significant difference was seen in the extent of thermal damage on either canine cornea or mouse dermis for the micropulse durations studied at either wavelength. The final set of experiments involved the use of mass spectrometry to determine whether amide bond breakage could occur in the proteins present in tissue as a result of direct absorptions of mid-infrared light into the amide I and amide II absorption bands. This analysis showed that there was no amide bond breakage due to irradiation at 6.45 microm on protein. PMID- 15815103 TI - Analysis of cardiac ventricular wall motion based on a three-dimensional electromechanical biventricular model. AB - This paper describes a biventricular model, which couples the electrical and mechanical properties of the heart, and computer simulations of ventricular wall motion and deformation by means of a biventricular model. In the constructed electromechanical model, the mechanical analysis was based on composite material theory and the finite-element method; the propagation of electrical excitation was simulated using an electrical heart model, and the resulting active forces were used to calculate ventricular wall motion. Regional deformation and Lagrangian strain tensors were calculated during the systole phase. Displacements, minimum principal strains and torsion angle were used to describe the motion of the two ventricles. The simulations showed that during the period of systole, (1) the right ventricular free wall moves towards the septum, and at the same time, the base and middle of the free wall move towards the apex, which reduces the volume of the right ventricle; the minimum principle strain (E3) is largest at the apex, then at the middle of the free wall and its direction is in the approximate direction of the epicardial muscle fibres; (2) the base and middle of the left ventricular free wall move towards the apex and the apex remains almost static; the torsion angle is largest at the apex; the minimum principle strain E3 is largest at the apex and its direction on the surface of the middle wall of the left ventricle is roughly in the fibre orientation. These results are in good accordance with results obtained from MR tagging images reported in the literature. This study suggests that such an electromechanical biventricular model has the potential to be used to assess the mechanical function of the two ventricles, and also could improve the accuracy of ECG simulation when it is used in heart-torso model-based body surface potential simulation studies. PMID- 15815104 TI - Direct thermal dose control of constrained focused ultrasound treatments: phantom and in vivo evaluation. AB - The first treatment control system that explicitly and automatically balances the efficacy and safety goals of noninvasive thermal therapies is described, and its performance is evaluated in phantoms and in vivo using ultrasound heating with a fixed, focused transducer. The treatment efficacy is quantified in terms of thermal dose delivered to the target. The developed feedback thermal dose controller has a cascade structure with the main nonlinear dose controller continuously generating the reference temperature trajectory for the secondary, constrained, model predictive temperature controller. The control system ensures thermal safety of the normal tissue by automatically complying with user specified constraints on the maximum allowable normal tissue temperatures. To reflect hardware limitations and to prevent cavitation, constraints on the maximum transducer power can also be imposed. It is shown that the developed controller can be used to achieve the minimum-time delivery of the desired thermal dose to the target without violating safety constraints, which is a novel and clinically desirable feature. The developed controller is model based, and requires patient- and site-specific models for its operation. These models were obtained during pre-treatment identification experiments. In our implementation, predictive models, internally used by the automatic treatment controller, are dynamically updated each time new temperature measurements become available. The adaptability of internal models safeguards against adverse effects of modelling errors, and ensures robust performance of the control system in the presence of a priori unknown treatment disturbances. The successful validation with two experimental models of considerably different thermal and ultrasound properties suggests the applicability of the developed treatment control system to different anatomical sites. PMID- 15815105 TI - [Evaluation of the global cardiovascular absolute risk: the Progetto CUORE individual score]. AB - To identify individuals at high risk for cardiovascular disease, the function of the Progetto CUORE has been identified and the global cardiovascular risk score has been built using data from different cohorts enrolled in the North, Center and South of Italy between the 80s and the 90s, whose risk factors had been collected using standardised procedures. A follow-up with a median duration of 9.5 years for men and 8.0 years for women has been achieved. Age, systolic blood pressure, total cholesterol, HDL-cholesterol, smoking habit, diabetes and hypertension treatment have been included in the function; the first major coronary or cerebrovascular event was considered as endpoint; 10-year survival has been assessed both for men and women. Out of 20 647 people aged 35-69 years with no previous cardiovascular events, 971 major cardiovascular events (636 coronary and 335 cerebrovascular) have been identified and validated. Risk factors coefficients have been assessed using the Cox proportional hazard model separately for men and women. The individual score is easy to be applied by general practitioners and cardiologists in order to achieve a fast and objective evaluation of the absolute global cardiovascular risk in primary prevention. PMID- 15815106 TI - Evolutive algorithms for beat-by-beat estimation of left ventricular mechanics. AB - Traditional methods to evaluate the ventricular mechanics need intraventricular pressure and volume recordings for multiple variably loaded beats. To do this, a complex and invasive procedure must be applied, that may decrease the clinical use. To overcome this limitation, a method to estimate the ventricular mechanics beat-by-beat is presented, modeling the ventricular pressure-volume relationship with a time-varying elastance function. The ability of the genetic algorithms (GAs) as identification technique is exploited. Applying GAs on surrogated data simulating variably loading conditions, the parameters of the time-varying elastance function, considered a measure of the contractility of the myocardial fibers are identified. These single-beat estimates are highly correlated with the end systolic pressure-volume relationship slope obtained by conventional multiple beat analysis. The main advantage in using GAs for single beat analysis may lie, in the perspective of an use for in vivo investigations, both in their stochastic nature, and in the guaranteed better performance with respect to other search techniques on problems involving noisy signals. Future studies will approach the reduction in GAs computational costs, for a real time in vivo application. PMID- 15815107 TI - Levels of persistent toxic substances in the general population in Italy. AB - Human exposure to persistent toxic substances (PTS) occurs daily, mainly through diet. As a consequence of continuous exposure, and because of their biological persistence, PTS are virtually present in all individuals, stored in their fatty tissues. The exposure of the Italian general population to PTS is a fact of relevance from a public health perspective, because of the number of toxic effects associated to these compounds, possibly occurring even at the current background level of exposure. In spite of this, data on PTS concentrations in humans, considered the best dose metric to carry out an adequate risk assessment, are scarce, as shown by the overview of available information we hereafter present. The Convention of Stockholm on persistent organic pollutants (POP), a group of highly toxic PTS, has entered into force last May. This will oblige parties to develop national implementation plans, thus creating a new opportunity to develop more efficient policies to control POP and other PTS present in food, humans and the environment. PMID- 15815108 TI - [The role of high risk groups in environmental health research]. AB - High risk groups in environmental epidemiology are characterized by particularly high exposure levels with respect pollutant agents occurring in the various environmental matrices. An investigation of these groups may offer elements of interest also for the general population, which is exposed to lower levels of the agents at study, besides providing the evidence needed in order to implement appropriate remedial action. The discussion of four case-studies concerning populations resident in the neighbourhood of waste dump sites, power lines, incinerators and in areas with presence of fluoro-edenitic fibres, throws light on the rationale of studies on high risk groups. Special emphasis in placed on validity aspects, connections between epidemiologic studies and remediation interventions and ethical issues. PMID- 15815109 TI - [The pigeon tick, Argas reflexus, and hazard for human health]. AB - The massive presence of the pigeon (Columba livia f. domestica) in urban areas not only is cause of serious aesthetic damages as decline of squares, of churches, of statues and buildings, but often creates many hygienic and medical problems. The pigeon, in fact, can transmit a great number of pathogenic agents and also be infested by several ectoparasites. Among these, Argas reflexus (Acari: Argasidae), the well known pigeon tick, is the most important ectoparasite that can involve also human health. Buildings infested by A. reflexus and attacks by this tick species to man are frequently reported, with an increasing interest for human health. In fact the pigeon tick bites can induce anaphylactic reactions for the presence of allergens, and as vector, the tick can also transmit some pathogens. This review deals with actual knowledge on the distribution, eco-biology, vector competence, prevention and control of A. reflexus in Italy. PMID- 15815110 TI - [Italian public and non-governmental organisations related to HIV/AIDS infection and migrant populations]. AB - This work is the result of two surveys carried out to obtain an overview of the specific types of activities conducted by public institutions and non governmental organisations (NGO) with regard to HIV infection and AIDS among migrant populations in Italy. The results reveal that the types of services offered to migrant populations differ when comparing public institutions to NGOs. Public institutions mainly provide services related to medical-health issues, whereas NGOs focus on social assistance and acceptance. Both types of organisations report problems in communicating to migrant populations information on HIV infection, sexually transmitted diseases, prevention, and therapy, which is a critical issue in the relationship between the psycho-social-health operator and the foreigner. PMID- 15815111 TI - [Drugs detection in street samples]. AB - The various steps of the analytical process are taken into account with reference to procedural problems linked to sampling operations, too. In particular, these aspects are treated: sampling procedures related to the various aspects of street samples, representative sampling of big amounts of controlled drugs, procedural problems in sampling of small amounts of drugs, the main methods of concealing. Up-to-date methods of extraction and chromatographic analysis are described. In the end, chemical profiling methods used to link different seizures of controlled drugs are briefly discussed. PMID- 15815112 TI - [Injuries in minors: study in the Emergency Department]. AB - The purpose of this research is to analyse occupational, household, sport and traffic related injuries among adolescents aged 14 to 17, during the first half of the year 2000, in the city of L'Aquila (Italy). All the data have been collected from the files of the Emergency Department. The occurrences have been 557: therefore 86 out of 1000 inhabitants have suffered from the above listed injuries. Among these ones, sport related injuries show to be prevalent: car accidents, occasional, household and professional injuries follow. Traffic related and occupational injury rates increase in the eldest children. In fact the "risk profile" shows to be worse in younger children. The study shows a major risk in 14 year old children, it is therefore necessary to focus on prevention and improvement in the data standardization, collection and recording at the Emergency Departments. PMID- 15815113 TI - [Characteristics of unconventional medicine users and their households in Italy]. AB - The results presented in this paper are part of the largest survey on unconventional medicine conducted so far (more than 140 000 people). Families with high socio-economical levels had the highest probability of choosing unconventional medicine. The use is strongly related to geographic area probably due to different availability of structures able to provide these therapies. At individual level the choice to use unconventional medicine seems a "cultural" trend associated with healthy life styles, and with positive attitudes towards prevention. It is interesting also to underline that the use of unconventional medicine is more frequent among women. Almost all users of unconventional medicine (80%) declare to be in good health, and this can indicate that unconventional therapies are mostly used for not serious and self limited conditions. Furthermore, use of unconventional medicine is often associated with a greater use of conventional drugs and with a greater frequency of check-up. This suggests a "complementary", more than "alternative" use of this kind of therapies. PMID- 15815114 TI - Suicide in schizophrenia. What are we going to do about it? AB - Suicide is the single major cause of death among patients with schizophrenia. Follow-up studies have estimated that 10-13% of these patients die by suicide. Various studies have identified risk factors for suicide in schizophrenic patients which are of great value for prediction and prevention of self-inflicted death. Despite great efforts, both on the side of drug treatment and psychosocial strategies, the number of suicides among schizophrenic patients has remained unchanged. We focus on a number of issues that are of paramount importance for the understanding and prevention of suicide in these patients, pointing to the need of supporting programs addressed to families and to establish a valid doctor patient relationship with these patients both during consultation with psychiatrists or general practitioners and with medical staff during inpatient treatment. PMID- 15815115 TI - Suicidality in DSM IV cluster B personality disorders. An overview. AB - Personality disorders are a considerable risk factor for suicidal behavior. In psychological autopsy studies, individuals with personality disorders are frequently found among suicide victims. Suicidal attempts (which are a major risk factor for completed suicide) are also an important issue among affected patients. We performed careful MedLine, Excerpta Medica, PsycLit and PsycINFO searches from 1980 to 2004. Search terms were "suicide", "personality disorders", "mortality", "follow-up". We identified studies dealing with suicide and the following areas of interest: borderline personality disorder, narcissistic personality disorder, antisocial personality disorder, comorbidity (especially with affective disorder and substance abuse disorder) and risk factors for suicide in personality disorders. Despite the fact that comorbidity with DSM-IV Axis I disorders often impairs a correct judgment of suicidality in personality disorders, this overview showed that some personality disorders have a stronger link to suicide and that identifiable risk factors may be used for the development of preventive measures. Given the fact that personality disorders have high prevalence in the general population, prediction and prevention of suicide among these individuals is a problem of public health. PMID- 15815116 TI - [Neuropsychological and behavioural assessment in Alzheimer's disease]. AB - To assess the onset and subsequent course of cognitive, behavioural and neuropsychiatric symptoms of Alzheimer's dementia (AD) we considered the clinical course of these groups of symptoms in 100 patients with NINCDS-ADRDA AD, > 65 years from onset to exitus during the phases t0 (early), t1 (state), t2 (neurological) and t3 (medical). Memory, depressive (40%), anxious (30%), sexual (15%) disturbances are frequent during t0. During t1, memory disturbances worsen in 90% of patients, and attention deficit (46%) and difficulty in abstract thinking appear. During t2, eating disorders (80%), stereotypy (38%), and delusions (23%) appear; anxiety and attention deficit (74%) worsen. During t3, eating disorders (95%) and delusions (46%) increase; higher brain functions can no more be assessed through neuropsychological tests. We showed progressive deterioration of cognitive function and behaviour, and abrupt onset and rapid progress of neuropsychiatric and medical disturbances during AD. PMID- 15815117 TI - [Italian regulation on the protection of human subjects participating to clinical studies, from the Nineties to 2004]. AB - In chronological order, and in the light of bioethics principles, the authors describe the Italian regulation which concerns the protection of human subjects participating in clinical trials from 1990 to July 2004, the related institution of Ethical Committees and the adoption of the tool of the informed consent. The publication includes the ties that have connected the Italian regulations to the European one since the beginning. During such period, the reception of the Good Clinical Practice guidelines - which occurred in 1992 first, and finally in 1997 has led to the establishment and the fostering of such important institutions as well as to the shaping of a network of Ethical Committees working on clinical trials and coordinated at a central level. In this paper the authors examine in particular: clinical trials of medicinal products, of medical devices and of ionizing radiations. Some implications of ethics are also discussed. PMID- 15815118 TI - [Ethics and prevention: environmental and individual disparities]. AB - The complex interactions which exist between environmental variabilities, genetic susceptibility of population subgroups, and high individual variability for age, sex, gender, ethnicity and general status of health, acquire an ever-increasing bioethical significance. Different risk conditions caused by toxic environmental agents and environmental inequities and inequalities are increasingly evident. "Social determinants" of health increase the probability of health effects and an effective intervention of prevision and prevention for environmental pathologies is needed. The debate on environmental inequalities caused by cultural, social and economic factors and the uncertainty about possible prevention emphasize the limits of the "bio-medical model". Ethics with its further anthropological and philosophical considerations may strongly help to understand the relationship between environmental pollution and health. PMID- 15815119 TI - Employer-based health insurance: a time for change? PMID- 15815120 TI - Continuous quality improvement in medicine: validation of a potential role for medical specialty societies. AB - A rigorous evaluation of the effects of continuous quality improvement (CQI) on medical practice has not yet been achieved on a large, multicenter scale. The authors sought to test whether a low-intensity CQI intervention could be used to speed the adoption of two coronary artery bypass grafting process-of-care measures on a national level. The infrastructure of the Society of Thoracic Surgeons' National Cardiac Database was used as a CQI Platform in a prospective randomized trial of CQI conducted between January 2001 and July 2002. Preoperative beta-blockade and internal mammary artery grafting in patients aged >75 years were the care processes used. Three hundred fifty-nine National Cardiac Database sites were randomized into two intervention groups (beta blocker, n=124; internal mammary artery grafting, n=114) and one control group (n=114). Each intervention arm received measure-specific information, including a call to action to a physician leader; educational products; and periodic longitudinal, nationally benchmarked, site-specific feedback. Incorporation of the specific care process into everyday clinical practice at the intervention site was the main outcome measure. The analyses included a site-level analysis of differences between pre- and postintervention measure use and a hierarchical analysis using risk-adjustment for patient characteristics and accounting for clustering due to site. Use of beta blockers increased vs. control (Delta=7% vs. Delta=4%), significant at both the site level (p=0.04) and in the hierarchical analyses (p=0.0006). Internal mammary artery graft use also increased vs. control (Delta=9% vs. Delta=5%; p=0.20 and p=0.11, respectively). However, lower volume IMA sites showed significant improvement over lower volume control sites (Delta=14% vs. Delta=8%; p=0.02 for interaction). A multifaceted, physician-led, low-intensity effort can have an impact on the adoption of care processes into national practice. This Society CQI Platform is a potential model for large-scale quality improvement efforts across all disciplines of medicine. PMID- 15815121 TI - Diabetes mellitus and heart failure. AB - The metabolic abnormalities associated with diabetes mellitus result in macrovascular and microvascular complications in multiple organ systems; it is the cardiovascular impact that accounts for the greatest morbidity and mortality associated with this disease. Heart failure, both with reduced and preserved systolic function, is a major complication, arising from the frequent associations with coronary atherosclerosis, hypertension, and a specific heart muscle dysfunction (cardiomyopathy) that occurs independently of coronary artery disease. Hyperglycemia, insulin resistance, and hypertension, together with activation of both the circulating and the tissue renin-angiotensin-aldosterone systems, contribute to structural fibrosis and autonomic neuropathy. Thus, it becomes imperative to identify cardiac abnormalities early in the course of both type 1 and type 2 diabetes to allow early and aggressive intervention to control glucose and blood pressure and to normalize blood lipid profiles. Patients with diabetes should be treated to secondary prevention targets, including blood pressure less than 130/80 mm Hg and low-density lipoprotein cholesterol level less than 100 mg/dL. Angiotensin-converting enzyme inhibitors, angiotensin receptor blockers,beta blockers, calcium channel-blockers, statins, and aspirin have all been demonstrated to significantly reduce cardiovascular morbidity and mortality in patients with diabetes. PMID- 15815122 TI - Spiral CT for pulmonary embolism: the paradigm has shifted. AB - Computed tomography (CT) is rapidly becoming the first-line modality for imaging pulmonary embolism (PE). However, limitations for the accurate diagnosis of small peripheral emboli have prevented the unanimous embrace of CT as the new standard of reference for imaging PE, although the actual significance of isolated peripheral emboli is uncertain. At the same time, the high negative predictive value of CT pulmonary angiography for excluding clinically significant PE has been established. The introduction of multidetector-row spiral CT has greatly improved visualization of peripheral pulmonary arteries and detection of small emboli. Previous concerns regarding the use of spiral CT for the accurate diagnosis of peripheral pulmonary emboli should thus be overcome. Multidetector row spiral CT has become a widely available and cost-effective technology and appears to have surpassed other imaging modalities for the accurate detection of central and peripheral PE. In this review, the authors assess the current role of spiral CT in the diagnostic algorithm of PE. PMID- 15815123 TI - Assessing quality in the cardiac catheterization laboratory. AB - Quality assurance and improvement have increasingly been the focus of health care providers, third-party payers, and patients. Because cardiovascular procedures are common, easily identifiable with claims data, and account for a relatively large proportion of health care expenditures, particular attention has been paid to quality assurance in the setting of the diagnostic and interventional cardiac catheterization laboratory. The structure, process, and outcomes domains of quality measurement in the interventional laboratory involve the maintenance of volume standards, the availability of surgical backup, consistent tracking of procedural outcomes and complications so they can be compared with national standards, and the application of evidence-based therapy. Quality assurance i the diagnostic laboratory revolves around the clinical proficiency of the operators, the maintenance and management of catheterization laboratory equipment, and the presence of a continuous quality improvement program. The evolution of interventional equipment and techniques along with the establishment of national registries has led to a gradual improvement in the quality of percutaneous coronary intervention. Given the dynamic nature of cardiology, adaptable quality assurance and quality improvement programs will remain the foundation of successful catheterization labs. PMID- 15815124 TI - The fallout of terrorism on public health policy. PMID- 15815125 TI - Building a multidisciplinary program: what toilets, dogs, young turks, and cartoons can teach us. PMID- 15815126 TI - Staving off death. PMID- 15815127 TI - The Heart Center of Indiana: changing the way hearts are treated. AB - The owners of The Heart Center of Indiana are continuing their long history of pioneering efforts. As a 50/50 joint venture between physicians and a large health system, The Heart Center is one of a new breed of "hospitals of the future" and the first of its kind in Indiana. Early indications of outstanding clinical and financial outcomes and guest satisfaction point to long-term success for The Heart Center of Indiana. PMID- 15815128 TI - Papillary muscle rupture following myocardial infarction. PMID- 15815129 TI - Abnormally long QT may be QU interval. PMID- 15815130 TI - Conflict of interest: the Trojan horse of specialty hospital legislation. PMID- 15815131 TI - Myocardial reperfusion: its assessment and its relation to clinical outcomes. AB - It has become increasingly apparent that epicardial blood flow restoration is necessary, but not sufficient, to achieve optimal clinical outcomes in the setting of acute coronary syndromes. Indeed, clinical outcomes are strongly associated with myocardial perfusion. The diagnostic tools available to assess myocardial perfusion following acute myocardial infarction and their association with clinical outcomes are reviewed here. Many simple angiographic markers--the Thrombolysis in Myocardial Infarction (TIMI) count frame, TIMI myocardial perfusion grade, pulsatile flow--are all readily available measures and are strongly related to clinical outcomes. Other tools, while accurate, remain limited by their high cost, low availability, and complexity for routine use. PMID- 15815132 TI - Genetic basis for hypertrophic cardiomyopathy: implications for diagnosis and treatment. AB - Familial hypertrophic cardiomyopathy is a genetic disease defined by cardiac hypertrophy in the absence of an increased external load. It is the most common inherited cardiac disorder occurring in 1 in 500 individuals. Ten genes exhibiting over 200 mutations have been identified. However, about 75% are due to mutations in just three genes: e-myosin heavy chain, cardiac troponin T, and myosin binding protein-C. Certain phenotypes are more common with certain genes, such as the myosin binding protein-C gene, which induces the disease predominantly in the fifth or sixth decade of life. Genetic animal models in the mouse and rabbit have helped to elucidate the pathophysiology. The primary defect imparted by the specific mutation alters contractile function, which stimulates release of various growth factors that induce secondary cardiac hypertrophy and fibrosis. Placebo single-blinded studies in the mouse indicate that losartan reverses the phenotype; in the rabbit, simvastatin essentially reversed the phenotype after 12 weeks of therapy. Clinical trials are ongoing in human familial hypertrophic cardiomyopathy. PMID- 15815133 TI - Sophisticated tests for susceptibility to atherosclerosis: an added value for heart hospitals. AB - Coronary atherosclerosis is the result of multiple complex metabolic disorders acting alone or in combination. Metabolic cardiology has advanced considerably in the past decade and now offers cardiologists a sophisticated approach to treating the underlying cause(s) of coronary artery disease in patients seen in invasive cardiology practices. Disorders which have now been powerfully linked to coronary artery disease include the small low-density lipoprotein trait, impaired reverse cholesterol transport, homocysteinemia, insulin resistance, aspects of inflammation, and infectious burden. These disorders are common and alone, or in combination, are found in over 90% of patients with coronary artery disease. Treatments for these disorders are well elucidated and have been associated with cardiovascular benefit. Incorporation of a sophisticated metabolic atherosclerosis management unit into existing invasive cardiology practices offers the cardiologist and patient many benefits. The cardiologist obtains a powerful tool that complements invasive cardiology, provides patients with access to cutting-edge diagnosis and treatment, and provides a forum that allows incorporation of clinically valid advances in metabolic cardiovascular care in a time frame far shorter than the traditional consensus recommendation approach. Incorporation of such a clinical service is consistent with the recent American College of Cardiology focus on the "CardioTeam." PMID- 15815134 TI - Magnetic resonance coronary angiography. AB - Coronary artery disease is the leading cause of death in the United States. Catheter-based x-ray angiography is the current standard for diagnosis of coronary artery disease. Coronary angiography using cardiovascular magnetic resonance is a rapidly evolving technique that can noninvasively image the coronary arteries. The authors will discuss the basic concepts and techniques of coronary angiography with cardiovascular magnetic resonance and motion suppression. Most of the relevant clinical studies that validate the use of coronary angiography with cardiovascular magnetic resonance will be presented, and potential future developments will be described. Coronary angiography with cardiovascular magnetic resonance could become the gold standard for imaging of the coronary arteries. PMID- 15815135 TI - Beating heart surgery: does it make a difference? AB - Beating heart techniques were introduced into coronary artery bypass graft (CABG) surgery in the mid 1990s in an attempt to decrease the complications associated with the use of cardiopulmonary bypass. Significant advances in technique and technology, including suction stabilizers and exposure devices, have now allowed all coronary vessels to be routinely approached without the support of cardiopulmonary bypass, while maintaining hemodynamic stability. Currently, approximately 25% of all CABG is performed without cardiopulmonary bypass in the United States. An extensive body of literature attests to the strong interest in the technique and the outcomes available for analysis. Although randomized trials do not yet show a mortality benefit, they are all underpowered to do so. A review of large retrospective databases does appear to demonstrate a mortality benefit, but patient selection contributes bias. Strong evidence of benefit exists for blood loss/transfusion, postoperative renal failure, sternal infection, myocardial injury, extubation time, length of stay, and cost savings. Particular benefit is apparent in high-risk patient subgroups, including the elderly, reoperative patients, and those with significant comorbidities. Off-pump CABG does appear to make a difference in improved outcomes by maintaining the excellent results of traditional on-pump CABG, but with less perioperative complications. PMID- 15815136 TI - New approaches to redo-coronary bypass grafting. AB - Patients presenting with recurrent symptoms after coronary artery bypass graft surgery (CABG) continue to be a significant therapeutic challenge for the interventional cardiologist and cardiac surgeon. Percutaneous transcatheter angioplasty and repeat CABG, directed at vein graft degeneration and progression of native atherosclerotic disease, carry substantially worse outcomes than primary procedures. Recent minimally invasive, surgical advances using minimal access incisions, arterial conduits, and off-pump techniques are demonstrating improved hospital outcomes compared with conventional CABG. These sternotomy sparing, beating heart surgical approaches combined with recent advances in drug eluting stents to provide complete revascularization may provide the best cardiac event-free survival for symptomatic CABG patients in the future. PMID- 15815137 TI - Assessing coronary plaque burden and plaque vulnerability: atherosclerosis imaging with IVUS and emerging noninvasive modalities. AB - Despite significantly improved preventive and therapeutic options, coronary artery disease remains the major cause of mortality in North America. A large number of acute coronary events, including acute myocardial infarction and sudden cardiac death, occur in patients without prior symptoms because these events are initiated by sudden rupture of mildly stenotic but vulnerable lesions. Recent results demonstrate a high prevalence of such plaques many years before clinical events occur. Because these lesions are mildly stenotic before the event, the angiographic evaluation of the lumen is not sufficient for their detection. Therefore, direct observation of coronary plaque burden and plaque vulnerability with in vivo tomographic imaging modalities is increasingly utilized. Intravascular ultrasound is the most established invasive method with a long clinical track record. More recently, noninvasive modalities including computed tomography and magnetic resonance imaging are emerging. PMID- 15815138 TI - The level I cardiovascular center: is it time? AB - There is no uniform approach to treating the 1.5 million US citizens who have an acute myocardial infarction (AMI) each year. This contrasts with the trauma system developed to efficiently triage and treat the critically injured accident victim. Only two thirds of patients with ST-segment elevation AMI in the United States are treated with thrombolytic therapy or primary angioplasty (percutaneous coronary intervention [PCI]) which can reduce the 30-day mortality rate from approximately 15% to 6%-10%. The Early Retavase-Thrombolysis in Myocardial Infarction (ER-TIMI) 19 trial demonstrated that AMI patients who received prehospital thrombolytic therapy and were brought to the nearest receiving hospital experienced a 32-minute reduction in the time to treatment and time to ST-elevation resolution compared with those treated at their time of hospital arrival. This expedited therapy was associated with a low in hospital mortality rate (4.7%). The potential benefit of facilitated PCI with partial-dose thrombolysis and abciximab administration was demonstrated by the Strategies for Patency Enhancement in the Emergency Department (SPEED) investigators who found that double bolus recombinant plasminogen activator (reteplase) (5 + 5 megaunits) and abciximab with the addition of early PCI, resulted in a final infarct-related artery TIMI 3 flow rate of 86% compared with 77% with combination therapy alone. The Primary Angioplasty in Acute Myocardial Infarction (PAMI) investigators have shown that patients admitted with infarct-related artery TIMI 3 flow at the time of primary PCI had less than a 1% 6-month mortality. Treating AMI patients with prehospital, partial dose thrombolysis followed by immediate transport to a Level I cardiovascular center (bypassing the closest hospital if necessary) for facilitated infarct-related artery PCI has the potential to reduce the mortality in ST-elevation AMI patients from 6%-10% to less than 4% which could translate into saving approximately 500 lives per day in the United States. It is time to validate this strategy with a randomized clinical trial, the Prehospital Administration of Thrombolytic Therapy With Urgent Culprit Artery Revascularization trial (PATCAR). PMID- 15815139 TI - On the future of adult cardiac surgery. AB - Considerable uncertainty exists about the role that will be played by cardiac surgery for acquired heart disease in the coming decade. In many institutions the volume of surgery for ischemic heart disease has already been affected by advances in interventional cardiology; the impact of those advances can only increase with the inexorable advance of technology. Percutaneous catheter intervention for valvular heart disease is also being pursued aggressively. Yet, like so many medical and surgical specialties, cardiac surgery has been forced to accept reduced reimbursements, and was also the first (and in many states is still the only) specialty to be required to submit to public audits and arbitrary standards for surgery volumes and outcomes. Hospital support for cardiac surgery programs has become less generous, and inevitably, the specialty has lost much of its desirability as a career choice. There is considerable concern about the specialty's ability to continue attracting the best surgical talent to its residencies, and in each of the last 2 years, 20 of 140 training positions did not fill in the initial match. (The implications of this phenomenon are made more confusing by uncertainty about the number of trainees that will be needed in the next decade.). PMID- 15815140 TI - The Society of Thoracic Surgeons' National Cardiac Database: provider engagement in continuous quality improvement. PMID- 15815142 TI - Drug development and the heart hospital. AB - Conducting clinical trials to support the development of new pharmaceutical entities or devices offers specialized heart hospitals unique opportunities. The populations they serve with cardiovascular diagnoses provide a predictable reservoir for recruitment of patients into trials of cardiovascular drugs or devices. The later stages in new chemical entity development require hundreds to thousands of patients to achieve statistical power. This need provides opportunities for rapid screening and identification of patients who would potentially benefit from and be appropriate for study. Clinically sophisticated support personnel to facilitate the conduct of research trials are likely to be available in the staff structure of a heart hospital. Meticulous attention to patient safety and privacy, good clinical practice, and high-quality data collection are imperative. PMID- 15815141 TI - The Mid America Heart Institute: part 1. AB - The Mid America Heart Institute (MAHI) is one of the first and largest hospitals developed and designed specifically for cardiovascular care. The MAHI hybrid model, which is a partnership between the not-for-profit Saint Luke's Health System, an independent academic medical center, and a private practice physician group, has been extremely successful in providing high-quality patient care as well as developing strong educational and research programs. The Heart Institute has been the leader in providing cardiovascular care in the Kansas City region since its inception in 1975. Although challenges in the future are substantial, it is felt that the MAHI is in an excellent position to deal with the serious issues in health care because of the Heart Institute, its facility, organization, administration, dedicated medical and support staff, and its unique business model of physician management. In part I, the authors describe the background and infrastructure of the Heart Institute. In part II, cardiovascular research and benefits of physician management will be addressed. PMID- 15815143 TI - ST displacements: a striking optical illusion. PMID- 15815144 TI - Infectivity of West Nile/dengue chimeric viruses for West Nile and dengue mosquito vectors. AB - West Nile virus (WN), an agent of significant human and veterinary disease, is endemic in the Old World and rapidly spreading throughout the Americas. Vaccines are needed to halt the geographic expansion of this virus and prevent disease where it is established. However, to preclude introduction of a vaccine virus into the environment, a live attenuated WN vaccine should have low potential for transmission by mosquitoes. A chimeric WN vaccine candidate was previously generated by replacing the membrane and envelope structural protein genes of recombinant dengue type 4 virus (rDEN4) with those of WN; a derivative of this virus, WN/DEN4-3'delta30, contains a 30-nucleotide deletion in the 3' untranslated region. To assess the potential for transmission by mosquitoes of these vaccine candidates, the ability of each chimeric virus to infect the mosquito midgut, disseminate to the head, and pass into the saliva was compared to that of their wild-type parental WN and DEN4 viruses in three vector species. The WN/DEN4 chimeric viruses were significantly attenuated in both Culex tarsalis, a vector able to transmit WN but not dengue, and in Ae. aegypti, a vector able to transmit dengue but not WN. However, the chimeric viruses were as infectious as either wild-type virus for Ae. albopictus, a vector able to transmit both dengue and WN. These results indicate that chimerization caused a contraction in vector host range rather than universal attenuation for mosquitoes per se. This restriction in potential vectors renders it less likely that WN/DEN4 and WN/DEN4-3'delta30 would be transmitted from vaccinees to mosquitoes. PMID- 15815145 TI - Newly recognized hantaviruses associated with hantavirus pulmonary syndrome in northern Brazil: partial genetic characterization of viruses and serologic implication of likely reservoirs. AB - Following the occurrence of the first laboratory-confirmed cases of hantavirus pulmonary syndrome (HPS) in Maranhao State, Brazil, rodents were trapped and rodent materials screened by ELISA for antibodies to Sin Nombre and Andes hantaviruses. Antibody-positive samples were tested by RT-PCR, amplified products were sequenced, and phylogenetic trees were constructed for comparison with known hantaviruses. From 104 rodent blood samples collected (40 Bolomys lasiurus, 52 Holochilus sciureus, 12 Oligoryzomys fornesi, and one Proechimys guyannensis), 21 (20.2%) were antibody-positive (one B. lasiurus, five O. fornesi, and 15 H. sciureus). Hantavirus RNA was amplified by PCR from two O. fornesi and four H. sciureus. Viral sequencing identified two hantavirus genotypes. The genotype recovered from O. fornesi, is designated herein as Anajatuba (ANAJ) and the genotype recovered from H. sciureus is designated Rio Mearim (RIME). Phylogenetic analysis of a 643-nucleotide region of the N segment showed both viruses to be most closely related (94-96% nucleotide homology) to Rio Mamore virus, a virus associated with Oligoryzomys microtis in Bolivia and Peru, but not found in northern Brazil. O. fornesi was frequently captured in and around human dwellings. H. sciureus, is a semi-aquatic rodent captured only in remote areas rarely frequented by humans. PMID- 15815146 TI - Arbovirus circulation, temporal distribution, and abundance of mosquito species in two Carolina bay habitats. AB - Carolina bays, a type of geomorphic feature, may be important in the ecology of mosquito vectors in South Carolina. Their hydrology varies from wetland habitats with marked flooding/drying regimes to permanently flooded spring-fed lakes. Moreover, they possess characteristics that contribute to the support of a particularly abundant and diverse invertebrate fauna. Although it has been estimated that 2,700+ bays exist in South Carolina, approximately 97% have been altered; < or = 200 bays remain intact, and only 36 are protected by state-funded conservation projects. We conducted a study in two distinct Carolina bay habitats, Savage Bay Heritage Preserve (SBHP) and Woods Bay State Park (WBSP), from June 1997 to July 1998 to determine mosquito temporal distribution, species composition, and the occurrence of arbovirus activity. The largest mosquito collection was obtained at WBSP (n = 31,172) representing 25 species followed by SBHP (n = 3,940) with 24 species. Anopheles crucians complex were the most common species encountered in both bays. Two virus isolates were obtained from SBHP in 1997: Keystone (KEY) virus from Ochlerotatus atlanticus-tormentor and Cache Valley (CV) virus from Oc. canadensis canadensis. Twenty-nine (29) arbovirus positive pools were obtained from WBSP: 28 in 1997 and one in 1998. KEY virus was isolated from three pools of Oc. atlanticus-tormentor and Tensaw (TEN) virus was isolated from two pools of An. crucians complex; 10 isolates could not be identified with the sera available. Additionally, 14 pools of An. crucians complex tested positive for Eastern equine encephalitis (EEE) virus antigen. These represent the first record of KEY and CV viruses in South Carolina. Our data indicate the presence of high mosquito density and diversity in both Carolina bay habitats, which may be influenced, in part, by seasonal changes in their hydroperiods. The study of mosquito and arbovirus ecology in Carolina Bay habitats could provide more information on the transmission dynamics of arboviruses and its impact on human and animal arboviral disease occurrence in South Carolina. PMID- 15815147 TI - A survey of environmental contamination with ascarid ova, Wallingford, Connecticut. AB - Few studies have been conducted in the United States to quantify the potential risk associated with encountering zoonotic ascarid ova in the environment. In an effort to raise awareness and to better understand the risk of acquiring visceral larva migrans in south central Connecticut, this environmental survey was conducted to determine the prevalence of ascarid ova (Toxocara canis, Toxocara cati, Baylisascaris columnaris, and Baylisascaris procyonis) in public areas of Wallingford, Connecticut, to compare prevalence levels among these public areas, and to determine what host species are primarily responsible for environmental contamination. A preliminary study was conducted to determine if ascarid ova of different species could be identified by size and appearance utilizing light microscopy alone; results did not support the differentiation of species via these methods. To determine the prevalence of environmental contamination with ascarid ova, samples of approximately 250 g of soil were collected from park green areas, playgrounds, public housing areas, parkways, and a school. Ova were detected in 46 (14.4%) of 319 samples collected. Ova were collected from three of the 60 (5.0%) park green area samples, 11 of the 40 (27.5%) playground samples, six of the 98 (6.1%) public housing samples, and 26 of the 96 (27.1%) parkway samples. Public areas of Wallingford, Connecticut are frequently contaminated by potentially infectious ascarid ova. Of particular concern is the high degree of contamination of playgrounds and the potential risk these areas pose to children's health. PMID- 15815148 TI - A comparision of West Nile Virus transmission by Ochlerotatus trivittatus (COQ.), Culex pipiens (L.), and Aedes albopictus (Skuse). AB - Transmission of West Nile virus (WNV) by Ochlerotatus trivittatus, Culex pipiens, and Aedes albopictus were compared 14 days after taking blood meals from viremic chickens with titers ranging from 10(2.5) to 10(9.5) cell infective dose (50)s (CID50s)/mL serum. Transmission occurred in one of four (25%) Oc. trivittatus and one of 25 (4%) Cx. pipiens that fed on chickens with titers of 10(5.5) CID50s/mL. No transmission occurred among two of 16 (13%) Oc. trivittatus or one of 25 (4%) Cx. pipiens that became infected after blood meals with titers of 10(5.0) and 10(4.5) CID50s/mL, the next lowest blood meal titers evaluated. Seventeen of 28 (61%) Ae. albopictus transmitted WNV after blood meals with titers of 10(7.0) CID50s/mL, but no infection or transmission was observed among 21 Ae. albopictus that fed on chickens with titers of 10(5.0) CID50s/mL, the next lowest titer evaluated. Transmission by all three species increased dramatically after blood meals with WNV titers of > or = 10(5.5) CID50s/mL. No significant differences occurred in dissemination and transmission rates of the three species after taking blood meals with titers of > 10(7.0) CID50s/mL. The cumulative mean +/- SE transmission rates of Oc. trivittatus, Cx. pipiens, and Ae. albopictus after blood meals with titers of > or = 10(7.0) CID50s/mL were 45.5 +/- 4.1%, 46.8 +/- 4.5%, and 72.4 +/- 5.5%. The cumulative mean dissemination rates of the three species were 78.3 +/- 6.7%, 74.8 +/- 2.6%, and 88.6 +/- 2.1%. The rates of transmission by the three species that developed disseminated infections after blood meals with titers of > or = 10(7.0) CID50s/mL were 58.8 +/- 4.4%, 62.6 +/- 5.8%, and 81.6 +/- 5.4%, respectively. In a previous study, we found that susceptibility of the three species to WNV was essentially the same when fed on chickens with WNV titers of > 10(7.0) CID50s/mL, but Oc. trivittatus and Cx. pipiens were more susceptible than Ae. albopictus to WNV at lower virus titers. The current study strongly suggests that Ae. albopictus is a more efficient vector than Oc. trivittatus and Cx. pipiens when fed blood meals with titers of > 10(7.0) CID50s/mL. However, Oc. trivittatus and Cx. pipiens might be more efficient as vectors when infected by blood meals with titers of < 10(7.0) CID50s/mL. PMID- 15815149 TI - Primary and secondary infection with Ehrlichia chaffeensis in white-tailed deer (Odocoileus virginianus). AB - White-tailed deer (Odocoileus virginianus) are the principal reservoir host for Ehrlichia chaffeensis, causative agent of human monocytic ehrlichiosis (HME). Because white-tailed deer maintain a long-term infection with E. chaffeensis and because deer can be naturally exposed to multiple strains of E. chaffeensis, we evaluated the response to secondary infection of E. chaffeensis in deer. For primary infection, six white-tailed deer were injected with 5.4 x 10(6) DH82 cells infected with the Arkansas strain of E. chaffeensis (Ark) and two control deer were injected with noninfected DH82 cells. On post-infection day 54, three E. chaffeensis (Ark) infected deer and one naive deer were injected with 4.2 x 10(6) cells infected with strain WTD-6045B E. chaffeensis, which differs from the Arkansas strain by number of nucleotide repeats in the variable length PCR target (VLPT) gene; three other Arkansas strain infected deer were injected with noninfected DH82 cells. All animals were monitored for 31 additional days. All deer in the primary infection became positive by PCR amplification of the 16S rRNA or VLPT genes and/or cell culture by DPI-8. PCR amplification of the VLPT gene on whole blood, cell culture, and tissues detected primary and/or secondary strains in all deer exposed to both primary and secondary strains; in one deer, the primary strain was cultured from the lymph node. Our culture results demonstrated that both strains were present; however, PCR detection suggests that the secondary strain may have been circulating in blood at higher levels. In conclusion, this study provides evidence that primary infection of deer with E. chaffeensis does not protect against subsequent exposure and confirms that deer can be simultaneously coinfected with at least two different strains of E. chaffeensis. PMID- 15815150 TI - Attempt to detect evidence for tick-borne encephalitis virus in ticks and mammalian wildlife in The Netherlands. AB - To investigate if tick-borne encephalitis virus (TBEV) is present in mammalian wildlife species or ticks in The Netherlands, serum samples and ticks were tested for TBEV antibodies and TBEV RNA, respectively. Serum samples were collected from wild boar (666), deer (13), fox (399), and rodents (90), and were tested for TBEV antibodies, using ELISA, and SN test or HI test. Over a period of 4 years, a total of 906 ticks was collected from seven regions in The Netherlands. In four different regions, this was done on a monthly basis and during four consecutive summers. All ticks were tested for TBEV RNA by RT-PCR. TBEV antibody was detected by ELISA in two (0.5%) sera of foxes and 49 (7%) sera of wild boar, but not confirmed by HI or SNT. TBEV RNA was not detected in any of 906 ticks. It was concluded that there is no real evidence for a TBEV reservoir in ticks or wildlife in The Netherlands. PMID- 15815151 TI - Argasid ticks as possible vectors of West Nile virus in Israel. AB - Mites and soft ticks collected directly from wild and domestic birds and their nests were tested for the presence of West Nile virus (WNV). The cattle egret argas, Argas arboreus, was collected from the nests of seven cattle egret colonies. Out of 1,000 A. arboreus pools examined, 16 were positive for WNV based on RT-PCR technique. The positive pools were from four nesting colonies of birds. Out of 37 cattle egret squabs examined, 37.8% had serum-neutralizing antibodies to WNV. WNV RNA was also detected in one out of 15 pools of R. turanicus, in one out of 21 pools of O. sylviarum, and in one out of 18 pools of D. gallinae, while 63 pools of A. reflexus, 11 of R. sanguineus, and 30 of Hyalomma spec. were negative. The role of mites and ticks in maintaining the endemic state of WNV in Israel is discussed. PMID- 15815152 TI - Triatoma dominicana sp. n. (Hemiptera: Reduviidae: Triatominae), and Trypanosoma antiquus sp. n. (Stercoraria: Trypanosomatidae), the first fossil evidence of a triatomine-trypanosomatid vector association. AB - Triatoma dominicana sp.n. (Hemiptera: Reduviidae: Triatominae) is described from a fifth instar nymph exuvia in Dominican amber and represents the first fossil representative of this subfamily. The new species is placed in the tribe Triatomini and tentatively in the genus Triatoma on the basis of the following characters; body length of 19.5 mm, body and appendages without long hairs, antenniferous tubercles without conspicuous apicolateral projections; abdomen with a series of five median tubercles; only fourth antennal segment annulated; lateral postocular callosities absent; stridulatory sulcus present and pronotum with discal and lateral tubercles. Trypanosoma antiquus sp. n. is described from two fecal droplets adjacent to Triatoma dominicana, thus providing the first fossil evidence of a triatomine-trypanosomatid vector association. The metatrypanosome stages of the fossil are similar in size to those of T. cruzi and could be a progenitor of this species. Mammalian hairs adjacent to the fecal droplets suggest that the vertebrate host for both the triatomine and trypanosomatid was a bat. Based on these findings, it is possible that bats were the original vertebrate hosts of T. cruzi-like trypanosomes. PMID- 15815153 TI - West Nile virus epizootiology in the southeastern United States, 2001. AB - We investigated mosquito and bird involvement in West Nile virus (WNV) transmission in July 2001 in Jefferson County, FL, and Lowndes County, GA. We detected 16 WNV-infected pools from Culex quinquefasciatus, Cx. salinarius, Cx. nigripalpus, and Culiseta melanura. In Florida, 11% of 353 bird sera neutralized WNV. Antibody prevalence was greatest in northern cardinal (Cardinalis cardinalis, 75%), northern mockingbird (Mimus polyglottus, 50%), common ground dove (Columbina passerina, 25%), common grackle (Quiscalus quiscula, 15%), domestic chicken (Gallus gallus, 16%), and house sparrow (Passer domesticus, 11%). Antibody-positive birds were detected in nine of 11 locations, among which prevalence in chickens ranged from 0% to 100%. Seropositive chickens were detected in Georgia as well. The primary transmission cycle of WNV in the southeastern United States apparently involves Culex mosquitoes and passerine birds. Chickens are frequently infected and may serve as effective sentinels in this region. PMID- 15815154 TI - Prevalence of antibodies to Coxiella burnetii among veterinary school dairy herds in the United States, 2003. AB - Prevalence of antibodies to Coxiella burnetii in 24 veterinary school-associated dairy herds in the United States was assessed through laboratory testing of bulk tank milk specimens by indirect immunofluorescent antibody assay. Twenty-two herds (92%) had evidence of antibodies to C. burnetii Phase I antibodies at a titer of > or = 1:16, and nine herds (38%) had Phase I antibody titers of > or = 1:256. These results suggest that C. burnetii infection is geographically widespread among dairy herds in the United States. PMID- 15815155 TI - Usefulness of a single item in a mail survey to identify persons with possible dementia: a new strategy for finding high-risk elders. AB - The objective of this study was to examine the characteristics of elderly persons who responded positively to a question about "severe memory problems" on a mailed health questionnaire yet were missed by the existing health risk algorithm to identify vulnerable elderly persons. A total of 324,471 respondents aged 65 and older completed a primary care health status questionnaire that gathered clinical information to quickly identify members with functional impairment, multiple chronic diseases, and higher medical care needs. The respondents were part of a large, integrated, not-for-profit managed care organization that implemented a model of care for elders using a uniform risk identification method across eight regions. Respondents with severe memory problems were compared to general respondents by morbidity, geriatric syndromes, functional impairments, service utilization, sensory impairments, sociodemographic characteristics, and activities of daily living. Of the respondents, 13,902 persons (4.3%) reported severe memory problems; the existing health risk algorithm missed 47.1% of these. When severe memory problems were included in the risk algorithm, identification increased from 11% to 13%, and risk prevalence by age groups ranged from 4.4% to 40.5%; one third had severe memory problems, a finding that was fairly consistent within age groups (28.4% to 36.5%). A question about severe memory problems should be incorporated into population risk-identification techniques. While false-negative rates are unknown, the false-positive rate of a self-report mail survey appears to be minimal. Persons reporting severe memory problems clearly have multiple comorbidities, higher prevalence of geriatric syndromes, and greater functional and sensory impairments. PMID- 15815156 TI - Specialty pharmacy: an emerging area of interest for medical management. AB - Specialty pharmaceuticals are expensive injectable and infusion therapies used to treat patients with chronic or life-threatening diseases. The high cost of these agents and their frequent usage in chronic diseases represent not only challenges, but also opportunities for medical management programs to improve the quality of care and moderate the rapid cost escalation seen in the industry. The number and variety of these agents have been increasing significantly, with hundreds of drug candidates in the development pipeline. The specialty pharmacy industry also is going through a consolidation stage, both horizontally and vertically. Industry approaches to medical management include the acquisition of specialty pharmacy companies, restrictive contracting to achieve concentrated buying power, and the development of utilization management strategies. PMID- 15815157 TI - A cohort study of the impact of a national disease management program on HEDIS diabetes outcomes. AB - Diabetes disease management programs (DDMP) are proliferating, but their overall impact in improving quality of care using Health Employer Data and Information Set (HEDIS) quality metrics has not been well studied. Furthermore, DDMPs are usually ongoing, but the incremental benefits of continuing the program beyond the initial patient educational intervention have not been rigorously tested. This study evaluates the impact of length of DDMP participation on diabetes related HEDIS 2002 quality indicators across 20 health plans. Results are stratified by duration of DDMP participation into three levels, "full participants" (6-12 months duration), "partial participants" (<6 months duration) and "non-participants" (0 months duration). The overall national compliance rate across all six combined HEDIS quality measures was 65.6% among full-participants (FP), 58.4% among partial-participants (PP) and 57.0% among non-participants (NP). This study demonstrates that participants in a comprehensive DDMP fair better than non-participants and that those with sustained participation (>6 months) benefit the most. PMID- 15815158 TI - Physician attitudes regarding cardiovascular risk reduction: the gaps between clinical importance, knowledge, and effectiveness. AB - Reducing risk factors for patients with vascular disease can reduce the subsequent incidence of cerebro-cardiovascular disease. While physicians have had extensive training in the importance of atherosclerotic vascular disease risk factor modification, evidence suggests that they systematically miss opportunities for clinical prevention during routine practice. The aim of this study was to identify whether physicians felt confident in their knowledge and effectiveness regarding counseling patients to reduce cardiovascular risk and to determine barriers to prevention interventions in the office setting. Surveys were mailed to 509 physicians affiliated with an academic community hospital. Nonrespondents were sent reminders and a second survey. Comparisons were made using chi-square analysis. Two hundred and five surveys were returned (40.3%). Thirty-six percent of physicians felt knowledgeable about weight management techniques, compared to 3% who were confident that they succeeded in their practice (p < 0.001). Similar patterns were found for Tobacco Cessation (62% versus 14%, p = 0.001), Alcohol Reduction (46% versus 7%, p < 0.001), Stress Management (35% versus 5%, p < 0.001), Exercise (53% versus 10%, p < 0.001), Nutrition (36% versus 8%, p < 0.001), Diabetes Management (48% versus 23%, p < 0.001), Blood Pressure Management (57% versus 43%, p < 0.001) and Lipid Management (59% versus 38%, p < 0.001). We identified a significant gap between physician confidence in their knowledge about risk factors and their effectiveness at providing counseling and obtaining results in their office. Most physicians felt that the routine office follow-up visit was an ineffective method for instituting vascular risk factor reduction. Alternate settings for risk factor reduction may be needed for improving atherosclerosis prevention. PMID- 15815159 TI - Prescribe by risk: the utility of a biomarker-based risk calculation in disease management to prevent heart disease. AB - Preventive treatment for those most at risk of heart disease rather than those with the highest blood pressure or cholesterol values may be a more efficacious strategy for disease management. This depends on accurate biomarker-based risk assessment tools. An evidence-based model of heart disease risk was developed using the Framingham model with an additional five risk factors, including three of the newer blood biomarkers. This was applied to the adult population of the 3rd National Health and Nutrition Examination Survey cohort. Additionally, the selection criteria for therapeutic intervention from the Adult Treatment Panel III guidelines (for hyperlipidemia) and the 7th Report of the Joint National Committee (for hypertension) were applied to the same subjects. Of this cohort 54% qualified for at least one of these medications while 18% qualified for both. Using this 18% cutoff, the 18% of the subjects with the highest calculated heart disease risk were also identified using the developed risk model. We applied established therapeutic reductions in heart disease probability to those identified by guidelines and to those identified by risk. Applying both drugs to the high-risk group (one third the size of the guidelines group) achieved the same reduction in population risk (about one fourth) as applying the drugs to the guideline groups and required only half as many prescriptions. Intermediate results were found when an intervention group was identified by a combination of both high risk and high levels of risk factors. In this simulation, identifying patients by heart disease risk level resulted in substantially fewer people being treated with fewer drugs and achieving a similar reduction in disease risk. PMID- 15815160 TI - Economic effectiveness of disease management programs: a meta-analysis. AB - The economic effectiveness of disease management programs, which are designed to improve the clinical and economic outcomes for chronically ill individuals, has been evaluated extensively. A literature search was performed with MEDLINE and other published sources for the period covering January 1995 to September 2003. The search was limited to empirical articles that measured the direct economic outcomes for asthma, diabetes, and heart disease management programs. Of the 360 articles and presentations evaluated, only 67 met the selection criteria for meta analysis, which included 32,041 subjects. Although some studies contained multiple measurements of direct economic outcomes, only one average effect size per study was included in the meta-analysis. Based on the studies included in the research, a meta-analysis provided a statistically significant answer to the question of whether disease management programs are economically effective. The magnitude of the observed average effect size for equally weighted studies was 0.311 (95% CI = 0.272-0.350). Statistically significant differences of effect sizes by study design, disease type and intensity of disease management program interventions were not found after a moderating variable, disease severity, was taken into consideration. The results suggest that disease management programs are more effective economically with severely ill enrollees and that chronic disease program interventions are most effective when coordinated with the overall level of disease severity. The findings can be generalized, which may assist health care policy makers and practitioners in addressing the issue of providing economically effective care for the growing number of individuals with chronic illness. PMID- 15815161 TI - DMAA Quality and Research Initiative. PMID- 15815162 TI - Cretaceous Park? A commentary on microbial paleomics. PMID- 15815163 TI - Beyond the principle of plentitude: a review of terrestrial planet habitability. AB - We review recent work that directly or indirectly addresses the habitability of terrestrial (rocky) planets like the Earth. Habitability has been traditionally defined in terms of an orbital semimajor axis within a range known as the habitable zone, but it is also well known that the habitability of Earth is due to many other astrophysical, geological, and geochemical factors. We focus this review on (1) recent refinements to habitable zone calculations; (2) the formation and orbital stability of terrestrial planets; (3) the tempo and mode of geologic activity (e.g., plate tectonics) on terrestrial planets; (4) the delivery of water to terrestrial planets in the habitable zone; and (5) the acquisition and loss of terrestrial planet carbon and nitrogen, elements that constitute important atmospheric gases responsible for habitable conditions on Earth's surface as well as being the building blocks of the biosphere itself. Finally, we consider recent work on evidence for the earliest habitable environments and the appearance of life itself on our planet. Such evidence provides us with an important, if nominal, calibration point for our search for other habitable worlds. PMID- 15815164 TI - Effects of a simulated martian UV flux on the cyanobacterium, Chroococcidiopsis sp. 029. AB - Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2 phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars. PMID- 15815165 TI - Microbial survival: the paleome: a sedimentary genetic record of past microbial communities. AB - Molecular genetic methods were used to analyze the remnants of microbial ecosystems contained within an ancient oceanic microbial habitat that was recovered from a continental drilled core of black shale approximately 100 million years in age. Bacterial ribosomal RNA genes were vertically amplified from the six different depths of a black shale core associated with a phosphate rich stratum, defined as one of the mid-Cretaceous oceanic anoxic events (OAEs). Although the black shale core was recovered from a terrestrial coring effort, the recovered 16S rRNA gene sequences showed affinity to microbial communities previously seen in deep-sea sedimentary environments (i.e., the microbial assemblage was easily recognizable as a marine community). In particular, a number of 16S rRNA gene clones of oceanic sulfate-reducing bacteria within the delta-Proteobacteria predominated at the OAE layer. The recovered bacterial DNA signatures are consistent with the interpretation that the sequences are derived from the past microbial communities buried in either sea-bottom or subseafloor environments during the sedimentation process and, after ceasing growth, preserved until the present. PMID- 15815166 TI - Finding a second sample of life on earth. AB - If life emerges readily under Earth-like conditions, the possibility arises of multiple terrestrial genesis events. We seek to quantify the probability of this scenario using estimates of the Archean bombardment rate and the fact that life established itself fairly rapidly on Earth once conditions became favorable. We find a significant likelihood that at least one more sample of life, referred to here as alien life, may have emerged on Earth, and could have coexisted with known life. Indeed, it is difficult to rule out the possibility of extant alien life. We offer some suggestions for how an alternative sample of life might be detected. PMID- 15815178 TI - Developmental life stage and couples' experiences with prostate cancer: a review of the literature. AB - Prostate cancer affects men in all adult life stages. As couples age, they face developmental tasks specific to their age. The combination of disease-related stressors and ongoing developmental changes may negatively affect the dyad's adjustment to prostate cancer and, consequently, their quality of life (QOL). In spite of this, a life stage perspective has not been used to understand the impact of diagnosis and treatment on patients and their partners across the aging life span. The purpose of this literature review was to explore the relationship between developmental age and disease-specific issues that may affect a couple's QOL as they adapt to a prostate cancer illness. The stages of aging are examined in 3 phases: late middle age (50-64 years); the young-old (65-74); and the old old (75 years and older). More specifically, these 3 phases were addressed first by presenting the normative developmental challenges of each phase, then disease related issues from the perspective of the patient, and finally from the perspective of the spousal caregiver. The literature review found that few studies considered age as a relevant factor in the analysis of outcomes of treatment; however, some differences among the groups for both the patient and the caregiver were identified. Ages of participants in the various studies covered a large span of time (50-86 years); consequently, recommendations from these studies do not consider the effect of developmental challenges on the couple's ability to adapt to a prostate cancer diagnosis. Knowledge gaps and implications for research using a developmental approach are identified. PMID- 15815179 TI - A qualitative analysis of interviews of men with early stage prostate cancer: the Prostate Cancer Lifestyle Trial. AB - We conducted interviews with a subsample of men (n = 26) participating in the Prostate Cancer Lifestyle Trial (PCLT), who had biopsy-proven prostate cancer (Gleason grade <7; prostate-specific antigen 4-10 ng/mL) and chose "watchful waiting" after diagnosis. The PCLT aimed at improving diet, exercise, stress management practices, and group support within a peer community setting. The aims of this study were to examine psychological, emotional, spiritual, and social reactions to (a) a diagnosis of early stage prostate cancer and (b) participation in the PCLT. Qualitative analyses were completed using MAXqda. Results indicated that the cancer diagnosis was met with anxiety, distress, and denial, or a "bump in the road" attitude. There was some indication that conflict with partners was exacerbated by the diagnosis. A significant component of the lifestyle intervention was involvement in the peer community. This community provided opportunities to experience connections with others facing similar health difficulties. Men generally expressed positive attitudes (emotionally, physically, and spiritually) about the lifestyle change, and felt that their participation in the program contributed to feelings of hope, optimism, and fighting spirit. Additionally, participants frequently mentioned increased comfort with emotional expression. PMID- 15815180 TI - Racial differences in prostate cancer treatment outcomes: a systematic review. AB - Although differences in prostate cancer incidence and mortality between black and white men are widely accepted, the existence of racial differences in treatment outcomes remains controversial. We conducted a systematic review of racial differences in prostate cancer treatment outcomes. Systematic review of literature from 1992-2002 was conducted. Database searches were performed using the terms: "prostate cancer" (keyword) or "prostate neoplasm" (subject heading) + "blacks" (subject heading) or "blacks" (keyword) + "African-Americans" (subject heading or "African-Americans" (keyword). Two hundred fifty-eight relevant articles were identified; 29 fit the inclusion criteria. All but 3 were retrospective. Seven (24%) studies were conducted at Veterans Affairs medical centers. Treatment included radical prostatectomy (15 studies), hormonal therapy (5 studies), and radiotherapy (12 studies). Three studies included more than 1 treatment. Twenty-three (79%) studies, observed no significant difference in treatment outcomes between races. The remainder found worse outcomes among black men, including worse 5-year survival (HR range, 2.35-96.74) and higher rates of PSA failure (OR range, 1.15-1.69). Most studies investigating racial differences in prostate cancer treatment outcomes over the past 10 years found no difference between races after controlling for tumor and patient characteristics. Efforts to narrow the gap between black and white prostate cancer mortality should focus on ensuring that all patients receive optimal treatment and that all patients become informed about the use of screening for early cancer detection. Research should focus on interventions to reduce advanced presentation of the disease and disease related mortality among black men. PMID- 15815181 TI - Predictors of breast and cervical screening in Vietnamese women in Harris County, Houston, Texas. AB - The Vietnamese are a quickly growing, important part of the Texas population. Breast cancer is known to have different biologic characteristics in Vietnamese women. In order to develop appropriate intervention and screening strategies, we conducted a study of barriers to cervical and breast screening in Vietnamese women in Harris County, Tex. Our objective was to characterize the demographic factors, beliefs, and barriers to cervical and breast cancer screening in our study population and test the effect of these on Papanicolaou test, breast self examination (BSE), medical breast examination (MBE), and mammography use. The Health Belief Model Scales for Measuring Beliefs Related to Breast Cancer (Champion VL, Nursing Research 1993;42:139-143) was the framework used to assess attitudes regarding risk of breast cancer and to design a component assessing risk of cervical cancer. The questionnaire addressed susceptibility, seriousness, benefits, barriers, and health about screening for breast and cervical cancer. It was translated into Vietnamese and back-translated into English prior to use. The questionnaire was mailed to Harris County residents. Those returned were entered into a database. The data were analyzed for validity using Chronbach's alpha. Simple descriptive analyses and nominal logistic regression identified predictors of Papanicolaou test, BSE, MBE, and mammography use. Twelve hundred surveys were mailed out to Vietnamese women using the telephone directory and the church directories in Harris County; 209 were returned and entered into the database. Of the respondents, 67% had ever received a Papanicolaou test; of these, 89% had received a Papanicolaou test within the past year; 55% of respondents had performed a BSE, 45% of patients received an MBE, and 45% of respondents had ever received a mammogram (15% of respondents had a mammogram during the previous year). The most significant predictors of Papanicolaou test, BSE, MBE, and mammography use were marital status (being married), high educational level, lack of barriers, a family history of the cancer, older age, and increased perception of seriousness. Compared to other studies of Vietnamese women, the women in Texas are among the highest users of the Papanicolaou test, BSE, MBE, and mammography. Barriers and incentives to breast and cervical screening were similar to those in other studies. PMID- 15815183 TI - A comparison of the information needs of women newly diagnosed with breast cancer in Malaysia and the United kingdom. AB - Little is known about the information needs of women with breast cancer in non Western societies. This study examined the priority information needs of 100 women with breast cancer in Malaysia and compared the findings to previous work involving 150 women diagnosed with breast cancer in the United Kingdom. The study used a valid and reliable measure, the Information Needs Questionnaire (INQ). The INQ contained 9 items of information related to physical, psychological, and social care, used successfully in Canada and the United Kingdom. The INQ was shown to have cross-cultural relevance and sensitivity. For Malaysian women, information about likelihood of cure, sexual attractiveness, and spread of disease were the most important information needs. For UK women, similar priorities were evident, apart from the item on sexual attractiveness, which was ranked much lower by women in the United Kingdom. The cultural similarities and differences that emerged from this study have implications for nurses in the cancer field caring for people from a diversity of cultural backgrounds. Breast care nurses are not a feature of the Malaysian healthcare system, although the findings from this study support the view that specialist nurses have a vital role to play in meeting the psychosocial needs of women with breast cancer in non Western societies. PMID- 15815184 TI - Evaluation of a Danish teaching program in breast self-examination. AB - This study investigated the effect of a BSE training program on women's knowledge, attitudes, and behavior regarding BSE. Postal questionnaires were sent to 629 women who had participated in 1998-2000 in the BSE training program run by Ribe County, Denmark, and to a local matched control group of the same size selected through personal registration numbers. Response rates were 77% and 56%, respectively. A significantly greater number of women who had attended BSE training reported that they knew how and when to do BSE, and what they should do if they discovered breast changes (97% compared to 66% in control group). Similar proportions in each group felt confident of finding any breast changes (57%) and believed that routine BSE can influence the chances of recovery from breast cancer (90%). There were also no significant differences between the intervention and control groups in the reporting of anxiety as a result of performing BSE (24% and 17%, respectively). The intervention group was significantly more likely to perform BSE regularly (66% compared to 52% in control group) and to use a more correct technique (44% compared to 20% in control group). It was concluded that a formal training program increases the likelihood of regular BSE performance with a correct technique. PMID- 15815185 TI - Quality of life 5 or more years post-autologous hematopoietic stem cell transplant. AB - This cross-sectional study used a mailed survey to evaluate the quality of life (QOL) of individuals at least 5 years post-autologous stem cell transplant and to determine instrument preference. Instruments selected were the Medical Outcomes Study-Short Form (MOS-SF-36) as the generic measure and the City of Hope-Quality of Life-Bone Marrow Transplant (COH-BMT) and the Functional Assessment of Cancer Therapy-Bone Marrow Transplant (FACT-BMT) as transplant-specific measures. Subjects received the MOS-SF-36 and were randomized to receive (1) COH-BMT, (2) FACT-BMT, or (3) COH-BMT and FACT-BMT. Ninety-two subjects returned completed forms, for a 56% response rate. A study-specific form indicated subjects preferred the BMT-specific instruments. The health of the majority of subjects (85%) was similar to or somewhat better than what it was the previous year. Their MOS-SF-36 scores for physical functioning, role-physical, bodily pain, and general health subscales were lower than the values for the general population, but those for the other subscales were not significantly different. When compared to the data reported by Hann and colleagues for posttransplant in breast cancer, study subjects scored significantly lower on all scales except General Health and Mental Health. COH-BMT scores compared with those reported by Whedon and Ferrel (Semin Oncol Nurs. 1994;10:42-57) were higher for Physical Well-Being, Spiritual Well-Being, and Global QOL. FACT-BMT results compared with those reported by McQuellen et al (Bone Marrow Transplant. 1997;19:357-368) showed that Physical, Social/Family, Emotional, and Functional Scores were similar; only BMT scores were significantly different. Research is needed to determine when QOL plateaus and whether instrument preference changes over time. Awareness of long-term effects that affect QOL can guide program revisions and facilitate decisions regarding the need for supportive rehabilitative services. PMID- 15815186 TI - Patients' experience of ambulatory self-administration of pamidronate in multiple myeloma. AB - The aim was to explore ambulatory self-administration of Pamidronate (Self-A-Pam) from a patient perspective in patients with multiple myeloma. Pamidronate is normally administered once a month as an intravenous infusion over 2 to 4 hours. Twenty-one patients were included, of whom 13 (6 women, 7 men) with a median age of 56 years (range 37-70) completed the educational program and subsequent ambulatory Self-A-Pam. An RN at the hospital initiated the Pamidronate therapy (90 mg). The patients then left hospital and later, on completion, they disconnected the infusion, either alone or with the assistance of a relative or significant other. Interviews were used to collect information about the experiences during the course of the Self-A-Pam. In total, 12 patients were interviewed after 3 doses of Self-A-Pam. One patient declined to participate in the interview. A qualitative analysis of the textual data was performed. Five main categories were identified: decision concerning Self-A-Pam, information and education, sources of practical help or support, effects of Self-A-Pam, and feelings and activities in relation to place (hospital, home, or public place). All 13 patients who started on Self-A-Pam went through 3 courses of Self-A-Pam during the study period. Many patients reported a gain in feelings of freedom/independence and time saving. However, some patients reported insufficient education and feelings of anxiety associated with the responsibility of handling the venous access device. PMID- 15815190 TI - Proteomics in prostate cancer. AB - PURPOSE OF REVIEW: State-of-the-art proteomics technologies are currently being assessed for utility in the study of prostatic malignancy. This review aims to provide background information on the current proteomics techniques employed in prostate cancer research, recent reports showing the potential application of proteomics in urological practice, and the future direction of proteomics in prostate cancer research and management. RECENT FINDINGS: Proteomic profiling of serum as a diagnostic tool and a platform for biomarker discovery in prostate cancer continues to draw favorable attention as well as close scrutiny as technological enhancements and multi-center study results are reported. In-vitro studies on prostate cell lines provide positive proof-of-principle results. The application of proteomics to query prostate tissue specimens yields novel prostate cancer biomarkers requiring further validation. The integration of proteomics with immunology also yields promising findings that may translate into clinically relevant biological assays. SUMMARY: The study of proteomics is an emerging research field, and current studies continue to display potential future usage in prostate cancer management. Succeeding scientific investigations will probably yield new diagnostic and prognostic tools for prostate cancer, provide insights into its underlying biology, and contribute to the development of novel treatment strategies. PMID- 15815191 TI - Update in the management of patients with hormone-refractory prostate cancer. AB - PURPOSE OF REVIEW: 2004 was a critical year for advances in prostate cancer treatment. The results from two pivotal multicenter phase III randomized studies are the first to demonstrate a survival benefit associated with chemotherapeutic treatment interventions in patients with hormone-refractory prostate cancer. This review will focus on an interpretation of the data from these two studies, the emerging role for chemotherapy in 2005 and beyond, and ongoing areas of clinical research. RECENT FINDINGS: Phase I and II studies have demonstrated biochemical and objective responses achieved with docetaxel-based chemotherapy in men with hormone-refractory prostate cancer. Two pivotal phase III clinical trials, TAX 327 and SWOG 9916 have demonstrated a survival advantage of docetaxel-based chemotherapy over mitoxantrone. Novel targeted therapies under investigation include calcitriol, growth factor-targeted agents, epothilones and others. SUMMARY: We now have a new standard of care for men with metastatic hormone refractory prostate cancer. Further investigation of docetaxel-based regimens in earlier clinical states of disease is warranted and may demonstrate greater clinical benefit. Additional chemotherapy agents are being studied, and may also add to the future armamentarium available for prostate cancer. The enrolment of patients into these studies is critical to the ongoing evolution of prostate cancer management. PMID- 15815192 TI - Update on magnetic resonance imaging, ProstaScint, and novel imaging in prostate cancer. AB - PURPOSE OF REVIEW: Despite marked stage migration, approximately a third of patients with clinically organ-confined prostate cancer will have extracapsular extension on pathological analysis. To improve outcomes further, alternative modalities of determining extraprostatic disease must be investigated beyond the current clinical parameters of digital rectal examination, prostate-specific antigen, and Gleason score. In addition, discerning the location of tumor in patients with biochemical recurrence directly affects treatment choice. This review highlights developments in prostate cancer imaging that may improve staging and treatment planning for prostate cancer patients. RECENT FINDINGS: Magnetic resonance imaging is most useful in the initial evaluation of men with prostate cancer. Its high specificity for the diagnosis of extracapsular extension is tempered by its low sensitivity. Positron emission tomography is used to assess regional and distant metastasis in many malignancies. Investigation into the most appropriate radiotracer for prostate cancer is paramount in any future applicability. The unique role of ProstaScint has been in evaluating patients with biochemical failure and in trying to discern metastasis before definitive therapy. The addition of spectroscopic imaging and nanoparticle infusion may lead to further improvements. Therefore, in differentiating local versus distant recurrence, appropriate interventions with radiotherapy or hormonal manipulation may be instituted. SUMMARY: Magnetic resonance imaging, ProstaScint, and to some extent positron emission tomography are all relevant imaging modalities for the evaluation of patients with prostate cancer. Current technological advances in each arena are improving the accuracy of diagnosis. Despite their specific limitations, use in the proper clinical setting may provide essential information to improve management decisions and disease outcomes. PMID- 15815193 TI - Relationship between obesity and prostate cancer. AB - PURPOSE OF REVIEW: This review examines the relationship between obesity and prostate cancer, with an update of recent research in this field. RECENT FINDINGS: A recent report of the Cancer Prevention Study II showed a direct relationship between increasing body mass index and prostate cancer mortality. However, the US Health Professionals Followup Study reported an inverse association between obesity and the risk of developing prostate cancer in men under 60 years of age or in those with a family history of prostate cancer. These studies illustrate the contradictory evidence linking obesity to prostate cancer risk and mortality. Body mass does not appear to affect the performance of prostate-specific antigen as a diagnostic test, and on prostate biopsy a lower body mass is associated with a higher cancer detection rate and a higher cancer volume as measured by core length involvement. In two recent radical prostatectomy series, obesity was associated with worse pathological features and higher biochemical recurrence rates. The higher risk of recurrence persisted in patients with organ-confined disease and negative surgical margins, implying that this risk is not related to surgical technique. Several potential biological mechanisms have been proposed to explain this link including hormonal alterations, hyperinsulinemia, glucose intolerance, and elevated insulin-like growth factor and leptin levels. SUMMARY: Recent literature provides evidence that obesity may promote the development of a more aggressive form of prostate cancer, resulting in higher recurrence rates after primary therapy and higher cancer mortality rates overall. The mechanism to explain the association between obesity and prostate cancer is unclear. PMID- 15815194 TI - Update on laparoscopic and robotic radical prostatectomy. AB - PURPOSE OF REVIEW: Laparoscopic and robotic-assisted prostatectomy have been proposed as alternatives to traditional open retropubic prostatectomy. In this review, we update the more recent data concerning the results, technical trends and controversies regarding these novel, minimally invasive procedures. RECENT FINDINGS: As a result of improved patient selection, a better understanding of surgical anatomy, and refinements in surgical techniques, traditional retropubic prostatectomy set the standards very high, leaving little room for improvement. In this review, the results of laparoscopic prostatectomy are compared with contemporary in addition to historical series. Besides the transperitoneal laparoscopic approach, which was almost exclusively used in the initial series, the introduction and development of the extraperitoneal laparoscopic approach meant a significant change in the surgical strategy of a number of teams worldwide. The relative merits of the transperitoneal and extraperitoneal approaches are discussed. Robotic radical prostatectomy is a promising technical innovation that allows us to overcome many of the inherent limitations of laparoscopic surgery. As a result of financial constraints, the experience has been limited to a few centres worldwide. SUMMARY: Although long-term results are still lacking, novel minimally invasive techniques seem to fulfil the highest standards of radical prostatectomy in terms of early oncological cure, functional results and morbidity. A standardization of data collection and evaluation methodology will be indispensable for a better comparison of the different series. PMID- 15815195 TI - Biochemical recurrence after definitive prostate cancer therapy. Part I: defining and localizing biochemical recurrence of prostate cancer. AB - PURPOSE OF REVIEW: The introduction of prostate-specific antigen into clinical practice heralded a dramatic shift in the epidemiology of prostate cancer. The diagnosis and treatment of lower stage disease in younger men with fewer competing co-morbidities has resulted in a longer period of post-treatment cancer surveillance and the potential for disease recurrence. Life-long periodic prostate-specific antigen testing for biochemical recurrence is standard of care; however, there is no single definition of biochemical recurrence that reliably predicts clinical recurrence. This review explores the complexities of biochemical recurrence, a thorough understanding of which is crucial to making appropriate treatment decisions after primary treatment. It also evaluates the array of diagnostic tests frequently employed when biochemical recurrence has occurred. RECENT FINDINGS: There is a disconnection between biochemical recurrence and progression to clinical disease. The definition of biochemical recurrence varies both by the prostate-specific antigen cut-point used and by the primary therapy employed. Furthermore, biochemical recurrence by itself appears not to be as reliable a predictor of eventual clinical recurrence as prostate specific antigen doubling time. Current imaging modalities are rarely useful in localizing disease when biochemical recurrence is first detected. SUMMARY: The correct interpretation of biochemical recurrence is crucial to treatment decision making. New data show that prostate-specific antigen doubling time during prostate-specific antigen recurrence may be a valid surrogate for death from the disease. The potential therefore exists for prostate-specific antigen doubling time to be accepted as a trial endpoint, which might accelerate drug approval by the United States Food and Drug Administration. PMID- 15815196 TI - Biochemical recurrence after definitive prostate cancer therapy. Part II: treatment strategies for biochemical recurrence of prostate cancer. AB - PURPOSE OF REVIEW: Through the prostate-specific antigen era, the proportion of men less than 55 years old with newly diagnosed prostate cancer more than doubled to almost 15%. As increasing numbers of men are living longer with prostate cancer, larger proportions will eventually present to our collective practices with rising prostate-specific antigen levels. Such prostate-specific antigen relapses, conservatively estimated to affect approximately 50 000 men each year, have become the most common form of advanced prostate cancer in the current period. RECENT FINDINGS: Increasing evidence suggests that early hormonal therapy improves progression-free survival and may alter the cancer-specific survival. However, there is a cost to pay in side-effects when androgen deprivation is administered over prolonged periods. The non-steroidal anti-androgen bicalutamide may offer an equivalent progression-free survival to castration without the complications of androgen deprivation. Observational data seem to indicate that high-risk individuals (i.e. those with high-grade, high-stage disease or a prostate-specific antigen doubling time less than 12 months) may also receive benefit from early therapy. SUMMARY: The definition of advanced prostate cancer has changed. Multimodal therapy improves cancer-specific outcomes especially in men with high-risk disease. The potential opportunities for novel therapeutic agents with low associated morbidity are great. PMID- 15815197 TI - Bibliography. Current world literature. Prostate cancer. PMID- 15815198 TI - Radiotherapy for esophageal cancer. PMID- 15815199 TI - Probiotics and disease: user friendly bacteria. PMID- 15815200 TI - Oral probiotics for maternal and newborn health. PMID- 15815201 TI - Diverticulitis: new concepts and new therapies. PMID- 15815202 TI - Dysphagia in aging. AB - Dysphagia is a common problem in older patients and is becoming a larger health care problem as the populations of the United States and other developed countries rapidly age. Changes in physiology with aging are seen in the upper esophageal sphincter and pharyngeal region in both symptomatic and asymptomatic older individuals. Age related changes in the esophageal body and lower esophageal sphincter are more difficult to identify, while esophageal sensation certainly is blunted with age. Stroke, Parkinson's disease, amyotrophic lateral sclerosis, Zenker's diverticula, and several other motility and structural disorders may cause oropharyngeal dysphagia in an older patient. Esophageal dysphagia can also be caused by both disorders of motility (achalasia, diffuse esophageal spasm, scleroderma and others) and structure (malignancy, strictures, rings, external compression, and others). Many of these disorders have an increased prevalence in older patients and should be sought with an appropriate diagnostic evaluation in older patients. The treatment of dysphagia in older patients is similar to that in younger patients, but more invasive therapies such as surgery may not be possible in some older patients making less aggressive medical and endoscopic therapy more attractive. PMID- 15815203 TI - A novel use of endoscopic clips in the treatment planning for radiation therapy (XRT) of esophageal cancer. AB - GOAL: To determine if the placement of metal endoclips improves radiotherapy (XRT) planning for esophageal cancer. BACKGROUND: XRT is an important modality in the treatment and palliation of esophageal cancer. Accurate simulation of the radiation field ensures optimal therapy while limiting toxicity. STUDY: Patients with esophageal carcinoma underwent EGD and endoscopic mucosal clip placement at the proximal and distal margins of the tumor. At XRT simulation the radiation oncologist simulated the field based on barium study, CT scan, and endoscopy report. A second radiation oncologist then assessed the ease of identification of the clips/margins of the tumor, shift in the field isocenter, and change in the radiation field width and length because of the metal clips. RESULTS: Seven patients with cancer of the esophagus had endoscopic clips placed followed by XRT simulation. The clips could be identified in all 7 patients. Simulation with the use of clips caused a shift in the field isocenter with a mean longitudinal shift (y-axis) of 3.5 cm (range, 1-6 cm) and a mean lateral shift (x-axis) of 0.33 cm (range, 0-1 cm). CONCLUSIONS: Endoscopically placed mucosal metal clips can aid field simulation when planning radiation therapy in patients with esophageal cancer. PMID- 15815204 TI - A follow-up study of patients with Zollinger-Ellison syndrome in the period 1966 2002: effects of surgical and medical treatments on long-term survival. AB - AIM: To evaluate the clinical history of a series of patients with Zollinger Ellison syndrome (ZES) in the period 1966 to 2002, before and after the introduction of the current antisecretive H2 receptor antagonists and proton pump inhibitors into clinical practice. PATIENTS AND METHODS: The study involved 18 ZES patients (9 males; mean age, 43 years; range, 12-70 years), 8 with Type 1 multiple endocrine neoplasia (MEN-1), diagnosed on the basis of standard criteria. We considered the type, number and effectiveness of surgical interventions before and after appropriate treatment, the localization of the gastrinoma, the presence of associated diseases, the causes of death, and the duration of survival. RESULTS: Total gastrectomy (but not antrectomy and vagotomy) and full compliance to antisecretory treatment reduced the number of operations from 29 to 9. One patient was cured (5.5%), whereas relapsing gastrinomas occurred in 4 patients and associated diseases or complications in ten. Death was related to ZES in 5 patients and to other causes in 4. CONCLUSIONS: Curing gastrinoma or appropriately inhibiting gastric acid hypersecretion in ZES patients prevent death and favors long-term survival, regardless of gastrin levels and the size or number of tumors. PMID- 15815205 TI - Is a 2-liter PEG preparation useful before capsule endoscopy? AB - AIMS: Small bowel contents can sometimes hamper the quality of capsule images. Our aim was to investigate the effect of PEG administered prior to capsule endoscopy (CE) upon quality of images, gastrointestinal transit time, and detection rate of small bowel bleeding lesions in patients with obscure gastrointestinal bleeding. PATIENTS AND METHODS: Forty-two consecutive patients were included. CE was performed following a 12-hour fasting period. The 16 first patients (Group A) received no preparation and the following 27 patients (Group B) received 2 L of PEG the night before. The quality of images was assessed at both in duodenojejunum and ileum level, using a scale including the presence of air bubbles, biliary secretion, and residue (1-4). RESULTS: Quality of images were not different in Group A compared with Group B in the duodenojejunum and in the ileum. Gastric transit time tended to be shorter in Group A compared with Group B (25.5 vs. 45.7 minutes) (P = 0.15), whereas small bowel transit was not different between both groups (271 vs. 288 minutes). Total small bowel CE examination was complete in Group A and in 24 of 26 in Group B (not significant). Potential bleeding lesions were seen in 8 patients in Group A and 12 in Group B (not significant). CONCLUSION: Our retrospective study suggests that 2 L PEG preparation seems able to improve neither the quality of CE images nor its diagnostic performance. Moreover, in our study, PEG tended to increase gastric emptying time and may constitute a limitation for small bowel complete examination. PMID- 15815206 TI - A randomized formula controlled trial of Bifidobacterium lactis and Streptococcus thermophilus for prevention of antibiotic-associated diarrhea in infants. AB - GOALS: This clinical trial was carried out to determine whether oral treatment with a commercial probiotic formula containing Bifidobacterium lactis and Streptococcus thermophilus would reduce the frequency of antibiotic-associated diarrhea (AAD) in infants. STUDY: In this double-bind formula controlled study, 80 infants, 6 to 36 months of age, were randomly assigned to receive a commercial formula containing 10 viable cells of B. lactis and 10 viable cells of S. thermophilus at the initiation of antibiotics for a duration of 15 days. The infants were assessed daily for formula intake, stool frequency, and stool consistency for a total duration of 30 days. Seventy-seven infants received nonsupplemented formula for the entire duration. RESULTS: There was a significant difference in the incidence of AAD in the children receiving probiotic supplemented formula (16%) than nonsupplemented formula (31%). CONCLUSIONS: The present study shows that prevention against AAD in infants was obtained by oral treatment with daily dose of B. lactis and S. thermophilus. PMID- 15815207 TI - Unemployment and disability in patients with moderately to severely active Crohn's disease. AB - GOALS: Unemployment and disability rates in Crohn's disease patients from the ACCENT I trial were assessed. Factors associated with employment and disability status were explored. BACKGROUND: Limited data regarding unemployment and disability status in patients with active Crohn's disease are available. STUDY: Baseline data were used to assess unemployment and disability rates. Logistic regression analysis examined factors that were associated with employment and disability status. Analysis of variance was used to compare quality of life. RESULTS: The baseline full-and part-time employment rates were 48% and 13%, respectively, with 39% of patients unemployed and 25% receiving disability compensation. Only 14% of 225 unemployed patients felt well enough to work if a job were available. Younger age, female gender, shorter disease duration, and prior bowel resection predicted a higher likelihood of unemployment. Younger age and female gender also predicted a higher likelihood of not being employed full time. Prior bowel resection predicted a higher likelihood of receiving disability compensation. Quality of life (Inflammatory Bowel Disease Questionnaire, Short Form-36) scores were significantly higher in employed patients. CONCLUSIONS: Patients with moderately to severely active Crohn's disease had low employment and high disability rates. Given their economic importance, assessment of these outcomes should be integrated into future evaluations of therapy, including clinical trials. PMID- 15815208 TI - Ileostomy adenocarcinomas in the setting of ulcerative colitis. AB - Adenocarcinomas arising at ileostomy sites in patients after colon resection for various diseases, such as ulcerative colitis (UC), familial adenomatous polyposis coli, and Crohn's disease, are rare occurrences but have been reported increasingly in the last 20 years. We report a case of adenocarcinoma arising in an ileostomy site in an 85-year-old woman with longstanding UC. She had pancolitis and underwent total proctocolectomy. Thirty-nine years later, her ileostomy site developed a granulation tissue-type lesion, which on initial biopsy revealed cytologic atypia in the presence of marked inflammation. A subsequent biopsy revealed adenocarcinoma with signet-ring cells and abundant extracellular mucin. Resection of the ileostomy was undertaken and a new ileostomy was performed. The literature on adenocarcinoma arising in the 23 patients with ulcerative colitis who received a Brooke or Kock ileostomy and had no prior history of neoplasm is reviewed. PMID- 15815209 TI - The diagnosis and management of benign hepatic tumors. AB - Benign hepatic tumors include a broad spectrum of regenerative and true neoplastic processes. Because of advances in imaging studies such as computed tomography (CT) and magnetic resonance imaging (MRI) as well as progress in immunohistochemistry, accurate diagnosis can now be made in a large percentage of patients without surgical laparotomy or resection. This article will focus on the pathogenesis, diagnosis, and management of focal benign lesions of the liver. Many of these tumors present with typical features in various imaging studies. On occasions, biopsies are required and/or surgical removal is needed. The most common benign hepatic tumors include cavernous hemangioma, focal nodular hyperplasia, hepatic adenoma, and nodular regenerative hyperplasia. In the majority of cases of benign hepatic tumors, patients are asymptomatic, and no treatment is indicated. The main indication for treatment is the presence of significant clinical symptoms or suspicion of malignancy or fear of malignant transformation. PMID- 15815210 TI - Endobiliary stents for palliation in patients with malignant obstructive jaundice. AB - BACKGROUND: Endobiliary drainage for malignant obstructive jaundice presents a viable palliative alternative. Its role and efficacy depend on factors related to the stent, procedure, and patient. GOALS: To review the evidence in the literature in which settings plastic or metal stents are cost-effective, and whether adjuvant measures or patient-related factors affect duration of stent patency. STUDY: Using databases a literature search was performed for papers published from 1979 to April 2004. All retrieved papers reporting experimental or clinical observations were rated according to strength of evidence, and carefully analyzed. RESULTS AND CONCLUSIONS: Metal stents (Wallstent) stay patent longer than plastic stents (large-bore polyethylene with side-holes), overall median 250 and 110 days, respectively, and seem cost-effective in patients with longer than about 6 months survival, which cannot be accurately predicted. Antibiotics or choleretic agents do not prolong stent patency in clinical settings. In case of stent occlusion, indicated stent exchanges and insertion of a plastic stent, respectively, seem cost-effective in patients initially treated with plastic and metal stents. PMID- 15815211 TI - Molecular epidemiology and clinical implications of TT virus (TTV) infection in Indian subjects. AB - GOALS: This study was aimed at obtaining data on the epidemiology and clinical course of TT virus (TTV) infections among Indian subjects. BACKGROUND: The TTV is a nonenveloped DNA virus, first identified in the peripheral blood of individuals with posttransfusion hepatitis of unknown etiology. There has been much conjecture regarding the disease association of this virus. STUDY: A total of 494 serum specimens from various groups of high-risk and control subjects were screened for TTV DNA by a semi-nested PCR, using the ORF1-derived N22 primers. The sera were also screened for the HBsAg surface antigen by an ELISA, HCV RNA by a 5' NCR-based RT-PCR and GBV-C/HGV RNA by a 5' UTR-based RT-PCR. The clinical and hepatic profiles of the various subjects were also studied. Seventy-one randomly picked TTV isolates were directly sequenced and their phylogeny was studied. RESULTS: TTV showed an overall positivity rate of 45.34% with a significant higher prevalence of 52.9% among the high-risk subjects as against a prevalence of 28% among healthy control subjects (P < 0.001). Abnormal liver function profiles were frequent among TTV viremic individuals and among the acute hepatitis cases studied a higher mortality rate correlated with a superimposed TTV infection. The 71 TTV isolates sequenced were found to belong to genotype 1a being closely homologous to TTV prototype TA278. CONCLUSION: The TT virus shows a significant prevalence in the Indian population, particularly among subjects at risk for acquiring parenterally transmitted infections. Our study corroborates a putative role of the virus in the etiology of liver disease, particularly in coinfection with other agents. PMID- 15815212 TI - Hereditary hemochromatosis: population screening based on phenotype in Brazilian blood donors. AB - GOALS: A population of blood donors was screened for hereditary hemochromatosis (HH) based on the phenotype strategy in accordance with the European consensus. STUDY: Nonfasting serum samples were obtained from 1,050 donors. Transferrin saturation (TS) was measured using a threshold of 45%. Donors with a TS > or = 45% were retested in a fasting sample. If TS was elevated, the participants were tested for iron overload by ferritin measurement followed by genetic testing. All donors underwent clinical and laboratory workup for expression of the disease. RESULTS: A total of 775 (74.6%) of the donors were men, 749 (72.1%) white, and had a mean age of 30 years (range, 8-60 years). Mean +/- SD TS was 25.9% +/- 13.1% (range, 2.1%-85.8%), and there were 58 (5.6%) donors with a TS > or = 45%. Fifty-four subjects had a repeat TS in a fasting serum sample with a mean +/- SD TS of 32.1% +/-16.1% (range, 15.4%-63.0%), and 12 donors had a TS > or = 45%. Ten complied with genetic testing and ferritin measurement. The study found four donors with HH-related mutations (C282Y and/or H63D); therefore, a gene allele frequency of 0.4%. Only the C282Y homozygote had diagnostic criteria for HH, defining a disease frequency of 0.1%. None of the donors who were mutations carriers had clinical or laboratory manifestations of organic injury. CONCLUSION: We conclude that this is a feasible screening strategy that, by timely diagnosing HH, allows patients not only to benefit from effective treatment but also to have disease progression halted. PMID- 15815213 TI - Beta blocker prophylaxis for patients with variceal hemorrhage. AB - GOALS: To characterize beta blocker therapy for the primary and secondary prevention of variceal hemorrhage. BACKGROUND: Variceal hemorrhage is one of the more frequent and severe complications of portal hypertension due to liver disease. Beta blocker therapy has been demonstrated to decrease risk of first bleed in patients with evidence of varices and recurrent bleeding and mortality in patients with history of prior variceal hemorrhage. STUDY: A total of 106 patients with liver disease hospitalized with suspected variceal hemorrhage were retrospectively reviewed. RESULTS: Half of patients had known varices, 44 (41.5%) of whom had experienced prior variceal hemorrhage. Only 21 (20%) were receiving beta blocker therapy at admission and 41 (48%) at discharge. The majority were not receiving therapy for primary prophylaxis (94%). Specific characteristics associated with beta blocker use could not be identified, although more patients with history of greater than two variceal hemorrhages were receiving beta blocker at admission (73% vs. 41%, P = 0.04) CONCLUSIONS: This study suggests that liver disease patients with varices are often not receiving beta blocker therapy to reduce risk of first or subsequent variceal hemorrhage. Opportunity exists to optimize use of this proven prophylactic treatment and bridge an apparent gap in standard of care. PMID- 15815214 TI - Nasobiliary tube management of postcholecystectomy bile leaks. AB - BACKGROUND: Endobiliary stenting is the traditional form of endoprosthetic drainage for biliary leaks. Nasobiliary tubes offer the advantage of easy removal and interval tube cholangiograms to assess leak resolution. AIM: To determine the efficacy of nasobiliary tube drainage in patients with postcholecystectomy biliary leaks and provide our experience with management of biliary leak using nasobiliary drains. MATERIALS AND METHODS: Retrospective study of 24 patients who were treated for postcholecystectomy biliary leaks in a tertiary referral center from 1998 to 2002. These patients were managed with either nasobiliary tube (NBT) alone or NBT + endoscopic sphincterotomy (ES). RESULTS: Twenty-four patients (mean age, 57.5 years; 50% women) had postcholecystectomy leak noted on ERCP. Twenty patients were managed by NBT+ES and 4 patients had NBT placement alone. In the NBT+ES group, 2 patients pulled their NBT out, but 18 patients had complete leak resolution in 3 to 9 days. In the NBT group, all patients had complete leak resolution in 4 to 12 days. Using an intention-to-treat analysis, 22 of 24 (92%) patients were successfully treated with NBT treatment over 3 to 12 days. CONCLUSIONS: ERCP with NBT placement is an effective and safe treatment modality in the management of postcholecystectomy biliary leaks. PMID- 15815216 TI - The statistical analysis of cramps. PMID- 15815217 TI - Probiotics affects vaginal flora in pregnant women, suggesting the possibility of preventing preterm labor. PMID- 15815218 TI - Recurrence of Helicobacter pylori infection after eradication therapy in Brazilian patients with peptic ulcer. PMID- 15815220 TI - Case management in South Korea from an American perspective. PMID- 15815219 TI - Necrotizing fasciitis secondary to intraabdominal infection in Crohn's disease. PMID- 15815221 TI - Beyond the chi-square: caregivers are more than just faceless statistics. PMID- 15815222 TI - A case management protocol: reducing unnecessary Medicare admissions in Florida. AB - The Florida Medicare quality improvement organization implemented a case management project with 20 acute care hospitals to reduce unnecessary Medicare admissions. The project called for hospitals to implement a protocol to assign patients to observation or inpatient admission status. Results of the project showed a 67% reduction in the denial rate for the participating facilities. This relative reduction in denial rates was nearly 3 times greater for the participating facilities as compared to the control facilities. The protocol can easily be adopted by hospitals. PMID- 15815224 TI - Global case management: the "nurse case manager" model applied to day surgery in Italy. AB - Day Surgery (DS) and prehospitalization emerged in Italy--as it had previously in the United States and other European countries--as able to improve healthcare and reduce costs. The purpose of this study was to evaluate the impact of the Nurse Case Manager (NCM) Model on patients undergoing DS procedures. The study assessed the intensity of pain, anxiety, and stress factors, and the level of discomfort experienced. The quasi-experimental study design involved two groups of patients (at two different DS hospital centers): one using the NCM model (Ward A) the other based on team nursing (Ward B). All patients undergoing DS for (a) varicose veins, (b) inguinal hernia, (c) breast biopsy, and (d) hemorrhoids were included. A valid questionnaire interview based on 22 questions was used (1) the morning before surgery, (2) the evening after surgery, and (3) 48 hr after the operation, by telephone. Of the 145 patients that were involved, 66 were from Ward A (45.5%) and 79 from Ward B (54.5%). When patient care was based on the NCM Model (Ward A), it seemed that (a) the pathway before DS was more efficient; (b) they perceived lower levels of perioperative anxiety and pain; and (c) they recognized the DS center as a unique point of reference for all postoperative problems. PMID- 15815225 TI - Nursing home transition: providing assistance to caregivers in transition program. AB - Transition from a skilled nursing facility or rehabilitation center to the community can be fragmented and insufficiently case managed, resulting in inadequate care recommendations, patient-caregiver distress, a delay in discharge, and a higher risk of nursing home readmission. The Providing Assistance to Caregivers in Transition (PACT) program is an interdisciplinary case management program designed to enhance nursing home discharge planning and case management support for the transitional period following a return to the community. During the PACT program's initial 24 months of operation, 38 of 42 opened cases were assisted in a discharge to the community. Of these, 30 remained at home for at least 6 months, 5 were readmitted within 6 months, and 3 others died. Caregivers reported satisfaction with instrumental (e.g., information about care options, facilitation of referrals to services) and emotional support. Nursing home cooperation was mixed. More work is needed to develop a broader referral base for the program. PMID- 15815226 TI - Case management curriculum in nursing education. AB - Case management has been viewed as an essential function within the current healthcare environment. While the importance of case management in providing quality patient care is documented in the literature and in statements by national nursing and healthcare organizations, there is limited identification and documentation of case management content in nursing education programs. The purpose of this study was to identify the current case management content taught in nursing education programs and to determine the essential concepts and skills of case management relevant to associate, baccalaureate, and graduate nursing education. Using the results of this study, one baccalaureate nursing program revised a chronic illness course to expand the classroom presentation and clinical experience associated with case management content. PMID- 15815227 TI - Case managers facilitate successful return to work in difficult cases utilizing return-to-work tools and therapist partnerships. PMID- 15815228 TI - Vocational rehabilitation: what the medical case manager needs to know. PMID- 15815229 TI - The anatomy of a readmission rate: are your information systems really counting what you think they are? PMID- 15815230 TI - Hepatitis C: laboratory tests to be done before the diagnosis can be made. PMID- 15815232 TI - Sufentanil, alfentanil, and fentanyl: impact on cerebrospinal fluid pressure in patients with brain tumors. AB - In order to evaluate the safety of the new synthetic opioids, alfentanil and sufentanil, in neurosurgical patients, we administered sufentanil 1 microg/kg i.v., alfentanil 50 microg/kg i.v. followed by an infusion of 1 microg/kg/min, or fentanyl 5 microg/kg i.v. to 30 patients with supratentorial tumors anesthetized with nitrous oxide (N2O), 60% in O2. Lumbar cerebrospinal fluid pressure (CSFP) and mean arterial pressure (MAP) responses were recorded for 10 min thereafter, while ventilation was held constant [mean PaCO2 = 36.1 +/- 1.0 mm Hg (SEM)]. There was no change in CSFP after fentanyl. In contrast, both sufentanil and alfentanil caused increases in CSFP, equal to 89 +/- 31 % SE (p < 0.05) and 22 +/ 5% (p < 0.05), respectively. MAP decreased after administration of each opioid. Peak decreases in cerebral perfusion pressure (MAP - CSFP) were 14 +/- 3% after fentanyl, 25 +/- 5% after sufentanil, and 37 +/- 3% after alfentanil. It is concluded that because sufentanil increased CSFP in patients who have brain tumors, it also may be contraindicated in other neurosurgical patients at risk for intracranial hypertension. Alfentanil may share this propensity, since CSFP increased despite a profound reduction in MAP. Among the three opioids evaluated, only fentanyl appears to be appropriate for supplementing N2O-2 anesthesia in patients who have compromised intracranial compliance. PMID- 15815233 TI - Cerebral blood volume and CSF pressure following administration of ketamine in dogs; modification by pre- or posttreatment with hypocapnia or diazepam. AB - The effects of ketamine (2 mg/kg) on cerebral blood (CBV) and cerebrospinal fluid (CSF) pressure, and the modification of ketamine's effects by pre- or posttreatment with hypocapnia (PaCO2 of 20-25 mm Hg) or diazepam (0.3 mg/kg) were determined in 26 dogs. In Group I (n = 7), the sequence of experimental treatments was as follows: control, ketamine, hypocapnia, and normocapnia. In Group II (n = 7), the sequence was control, hypocapnia, ketamine, and normocapnia. In Group III (n = 6), the sequence was control, ketamine, and diazepam. In Group IV In (n = 6), the sequence was control, diazepam, and ketamine. Ketamine alone increased CBV by 11.7 +/- 6.6 (Group III) to 12.4 +/- 6.9% (Group I) (mean +/- SD) and increased CSF pressure by 6.0 +/- 3.9 (Group I) to 10.7 +/- 5.2 (Group III) cm H2O. Pretreatment with hypocapnia or diazepam reduced CSF pressure and CBV so that when ketamine was administered after 15 min of hypocapnia or diazepam, CSF pressure and CBV rose only to prehypocapnia or prediazepam levels. When hypocapnia or diazepam were administered 15 min after ketamine, CSF pressure and CBV decreased to preketamine levels. For all treatments, CSF pressure changed in parallel with CBV in the absence of significant changes in PaCO2, expired CO2, or mean arterial blood pressure (except for a decrease of mean arterial blood pressure with diazepam pretreatment). We conclude the following: (a) ketamine increases CBV concomitant with increases in CSF pressure, (b) ketamine increases CBV and CSF pressure independent of significant change in PaCO2, and (c) pre- or posttreatment with hypocapnia or diazepam reverses ketamine-induced increase of CBV and CSF pressure in dogs with normal CSF pressure and no cerebrovascular damage. PMID- 15815234 TI - Effect of time and dose on scalp-recorded somatosensory evoked potential wave augmentation by etomidate. AB - Bolus etomidate transiently increases the amplitude of scalprecorded somatosensory evoked potentials (SSEPs). The reproducibility of this augmentation and its dose-response relationship are unknown. In unpremedicated patients, we studied the effect on the SSEP of repetitive administration of single doses of etomidate (0.1 mg/kg i.v. bolus) in six patients and increasing doses of etomidate in six additional patients. Anesthesia was induced with fentanyl (15-20 microg/kg i.v.) plus thiopental (1-2 mg/kg i.v.) and maintannined with 0.4-0.8% isoflurane in oxygen, and the surgical incision was infiltrated with bupivicaine (0.5% without epinephrine). Etomidate administration was delayed for 30 min following anesthesia induction. In group 1, 0.1 mg/kg etomidate was administered intravenously as a bolus three times at 30-min intervals. In group 2, 0.05, 0.1, or 0.2 mg/kg was administered at 30-min intervals in random order in each patient. SSEPs were measured immediately before and once each minute for 5 min after etomidate administration following nondominant median nerve stimulation. In group 1, administration of 0.1 mg/kg etomidate (three trials) increased latency of an early negative wave (N20; latency approximately 20 ms) and a positive wave following N20 (P23; latency approximately 23 ms) by 1.0-1.4 and 1.3-2.6 ms, respectively (p < 0.05). P15N20 amplitude was increased by approximately 50% (range 36-76%; p < 0.05) and N20P23 amplitude was increased to 174% of control (range 173-178%; p < 0.05) and the amplitude increase was similar during the three etomidate administrations for both P15N20 and N20P23. Latency remained elevated by approximately 1.5 ms and amplitude remained elevated (P15N20 = 138%; N20P23 = 150%) 5 min following injection. Mean arterial blood pressure was unchanged by 0.1 mg/kg etomidate. In group 2, 0.05 mg/kg etomidate altered neither amplitude nor latency. However, 0.1 and 0.2 mg/kg increased N20P23 amplitude to 161 +/- 33 and 230 +/- 10% of control (p < 0.05), respectively. N20 and P23 latency were increased by 0.1 mg/kg etomidate by approximately 1.0 ms, while 0.2 mg/kg increased N20 latency by 1.0 ms and P23 latency by 1.5 ms. Bolus administration of etomidate (0.1 mg/kg) reproducibly increased SSEP amplitude and a larger dose (0.2 mg/kg) further increased amplitude augmentation. Thus, intermittent injection of etomidate can be used to augment small SSEP waves with reproducible increases in wave amplitude. PMID- 15815235 TI - Cerebral metabolic depression and brain protection produced by midazolam and etomidate in the rat. AB - Midazolam and etomidate have been shown to depress cerebral metabolism and may protect the brain during ischemia. However, it has been reported that etomidate may produce EEG spiking activity and seizures, which could adversely affect outcome. We compared the effects of midazolam and etomidate on EEG, cerebral blood flow (CBF), and cerebral cortical oxygen consumption (CMRO2) as well as neurologic outcome following incomplete cerebral ischemia in the rat. CBF was measured with radioactive microspheres and cortical CMRO2 was calculated by multiplying cortical CBF by the arterial-sagittal sinus oxygen content. Incomplete ischemia was produced by unilateral carotid artery occlusion combined with hemorrhagic hypotension. In low doses (0.02 mg/kg/min i.v.), both midazolam and etomidate depressed EEG, decreased CMRO2, and improved outcome from ischemia compared to nitrous oxide control rats. At a higher dose (0.2 mg/kg/min i.v.), midazolam further depressed EEG and CMRO2 and again improved outcome compared to N2O controls. In contrast, high dose etomidate (0.2 mg/kg/min) produced spiking EEG activity without further depression of CMRO2 and a worsening of outcome following cerebral ischemia. These results support previous reports that midazolam and etomidate may protect the brain from incomplete cerebral ischemia but suggest that EEG spiking activity associated with high dose etomidate may be associated with a worse outcome. PMID- 15815236 TI - Hypocapnia prevents the decrease in regional cerebral metabolism during isoflurane-induced hypotension. AB - In neurologic surgery, induced hypotension is often used while the patient is hypocapnic. We investigated, by tissue biopsy methods and scintillation counting, the regional cerebral glucose utilization (rCMRglc) and blood flow (rCBF) in rats subjected to hypocapnia alone and in combination with hypotension. Anesthesia was maintained with 1.0% isoflurane in nitrous oxide/oxygen. Seven rats were maintained at PaCO2 of 40 mm Hg, six rats were ventilated to PaCO2 of 20 mm Hg, and six animals to PaCO2 of 20 mm Hg in combination with arterial hypotension of 50 mm Hg induced by isoflurane 2.5-3.5%. During hypocapnia, rCMRglc tended to increase in all regions, but the increase was statistically insignificant; rCBF was reduced uniformly by 40%. During combined hypocapnia/hypotension, rCMRglc was unaltered when compared to hypocapnia; compared to normocapnia, increases were seen in hippocampus and cerebellum. During hypocapnia/hypotension, rCBF was unaltered in cortical areas, while increases were seen in all subcortical areas compared to hypocapnia. Regional values of the ratio of rCBF/rCMRglc indicated that during hypocapnia and hypotension induced by isoflurane in nitrous oxide/oxygen, the individual brain areas were perfused according to their metabolic needs. It is suggested that hypocapnia may prevent the decrease in rCMRglc, which is usually observed during deep isoflurane anesthesia. PMID- 15815237 TI - Hemodynamic status following regional and general anesthesia for carotid endarterectomy. AB - We prospectively studied 23 patients undergoing carotid endarterectomy under regional (n = 13) or general (n = 10) anesthesia to determine the hemodynamic basis of increased frequency in the need for postoperative vasopressor support when regional anesthesia was used. Anesthesia and postoperative care were conducted without reference to hemodynamic data from pulmonary artery catheterization. Although mean arterial pressure was similar in the two groups postoperatively, 11 of the 13 patients undergoing regional anesthesia and 3 of the 10 patients undergoing general anesthesia required phenylephrine postoperatively. No patient required therapy postoperatively to reduce a systolic pressure exceeding 160 mm Hg. Mean arterial pressure remained below the preoperative baseline value in both groups (p < 0.05 with general anesthesia; p = 0.06 with regional anesthesia) during follow-up. In the general anesthesia group, systemic vascular resistance declined significantly below baseline (p < 0.05) following the operation, accompanied by a decline in mean arterial pressure (p < 0.05) and a higher cardiac output. Intraoperative fluid requirements were greater during general anesthesia than during regional anesthesia (p < 0.01). Pulmonary artery occlusion pressure was lower postoperatively than at baseline in both groups (p < 0.05). Pulmonary artery occlusion pressure was higher in the general anesthesia group despite the greater use of phenylephrine in the regional anesthesia group. PMID- 15815238 TI - The effects of regional and general anesthesia on blood pressure control after carotid endarterectomy. AB - We retrospectively reviewed the influence of preoperative blood pressure control and regional vs. general anesthetic techniques on the incidence of intraoperative and postoperative (recovery room and intensive care unit) hypotension and hypertension in 249 carotid endarterectomy patients. Preoperative blood pressure was classified as uncontrolled hypertension (systolic blood pressure >/= 170 mm Hg and/or diastolic blood pressure >/= 95 mm Hg), controlled hypertension (blood pressure <170/95 mm Hg on chronic antihypertensive therapy), or normotension (blood pressure <170/95 mm Hg without antihypertensive therapy). Hypotension, as defined by the requirement for vasopressor administration to maintain a systolic blood pressure of at least 120 mm Hg, occurred more frequently after regional than after general anesthesia (p < 0.05). Postoperative hypertension was defined as a systolic blood pressure >/= 200 mm Hg and/or a diastolic blood pressure >/= 110 mm Hg in the recovery room or in the Intensive Care Unit. Preoperative hypertension was not associated with acute postoperative hypertension in the intensive care unit in either the regional anesthesia (n = 190) or the general anesthesia (n = 59) groups, although with either type of anesthesia, preoperative hypertension was associated with an increased incidence of hypertension in the recovery room (p < 0.01 regional; p < 0.005 general). PMID- 15815239 TI - Flumazenil reversal of midazolam in dogs: dose-related changes in cerebral blood flow, metabolism, EEG, and CSF pressure. AB - Large doses of flumazenil, given rapidly (over 5-10 s), are reported to elevate cerebral blood flow (CBF) and intracranial pressure to supranormal values when given to dogs receiving midazolam. This study examined the cerebral effects of giving smaller, graduated doses of flumazenil (0.0025, 0.01, 0.04, and 0.16 mg/kg), slowly (over 60 s), to dogs receiving midazolam and to dogs not receiving midazolam both when cerebrospinal fluid (CSF) pressure was normal and when CSF pressure was elevated (intracranial balloon) to about 30 mm Hg. In dogs with normal CSF pressure that were receiving midazolam, the effects of flumazenil were as follows: (a) low doses of flumazenil caused reversal of the reduction in cerebral metabolic rate for oxygen (CMRO2) and activity of the electroencephalogram produced by midazolam, (b) moderate doses of flumazenil produced a decrease of cerebral vascular resistance, and an increase of CBF and CSF pressure that did not significantly change cerebral perfusion pressure (CPP), and (c) the highest dose of flumazenil increased CBF to supranormal values. All of these flumazenil effects "peaked" at 3-6 min, with values returning to pre flumazenil levels by 15-30 min. Flumazenil caused no such changes in dogs with elevated CSF pressure that were receiving midazolam or in dogs that were not receiving midazolam. The results are consistent with a specific, doserelated benzodiazepineantagonist action of flumazenil. Lack of flumazenil effect at elevated CSF pressure may reflect reversible changes in cerebral structure, metabolism, or benzodiazepine receptors produced by the intracranial balloon and elevation of CSF pressure. The doses of flumazenil used here to reverse the cerebral effects of midazolam appear unlikely to produce adverse effects because increase of CMRO2 was matched by increase of CBF, the mean increase of CSF pressure was modest (+9 +/- 3 mm Hg, mean +/- SEM), and CPP was unchanged. PMID- 15815240 TI - Thromboxane A2 receptor antagonist SQ29548 does not improve canine postischemic cerebral hypoperfusion. AB - The thromboxane receptor antagonist SQ29548 was administered to mongrel dogs after an 11-min period of global cerebral ischemia to test the hypothesis that it would improve delayed postischemic cerebral hypoperfusion (PIH) during concurrent elevations in cerebral venous thromboxane B2 (TxB2), the stable metabolite of thromboxane A2 (TxA2). Immediately following an 11-min period of aortic root cross-clamping, six dogs received 0.2 mg/kg of SQ29548 intravenously followed by continuous infusion of 0.2 mg/kg/h (TRA group) and six received a saline placebo (control). Over the next 120 min, cerebral venous outflow was measured from the confluence of the sagittal and lateral sinuses, while arterial and cerebral venous samples were obtained for measurement of TxB2. Delayed postischemic cerebral hypoperfusion, confirmed in the control group (p < 0.05) (16 +/- 2.7 vs. 32 +/- 2.4 ml/min baseline), was nearly identical in the TRA group (14 +/- 0.9 vs. 30 +/- 2.5 ml/min baseline). Cerebral venous TxB2 levels rose dramatically in both groups after ischemia (3670 +/- 440 vs. 1100 +/- 350 pg/ml baseline, control and 2720 +/- 170 vs. 580 +/- 100 pg/ml baseline, TRA group) (p < 0.05). There were no significant group mean differences in any of the other hemodynamic data except mean arterial pressure (MAP) at T120 and Hgb at baseline. We conclude that postischemic intravenous administration of the thromboxane receptor-specific antagonist SQ29548 fails to improve delayed postischemic cerebral hypoperfusion and does not alter cerebral venous TxB2 release in this canine model of global cerebral ischemia. PMID- 15815241 TI - Immediate hemodynamic management following subarachnoid hemorrhage during embolization of cerebral vascular abnormalities. AB - Three patients are reported who suffered intracranial hemmorrhage during interventionally neuroradiology procedures. All had severe hypertension, cardiac arrhythmias and loss of consciousness. Immediate CT scan showed small amounts of blood in all patients. Arrhythmias were controlled with lidocaine. In two patients, the hypertension was treated only with barbiturates plus fentanyl and in the third with labetol and nitroprusside. In the patient treated with nitroprusside and one patient with barbiturate plus fentanyl a 14-20 min period of hemodynamic stability was followed by vasomotor collapse requiring intravenous vasopressor support. The patient initially treated with nitroprusside suffered irreversibly brain injury whereas the other two improved neurologically following their intracranial bleed. These patients suggest that nitroprusside is detrimental in the acute treatment of malignant hypertension following intracranial hemorrhage. PMID- 15815243 TI - Therapeutic loading dose of phenytoin in anesthetized patients. PMID- 15815242 TI - Undiagnosed pheochromocytoma in a patient with lindau disease. AB - Lindau disease is characterized by angioblastic lesions of the cerebellum and spinal cord. It may, rarely, be associated with pheochromocytoma. We describe the management of a patient with Lindau disease who was scheduled to undergo craniotomy for removal of a cerebellar hemangioblastoma and who developed severe hypertension following induction of anesthesia. Subsequent work up revealed a previously undiagnosed pheochromocytoma. The importance of recognizing a possible association between these two conditions is discussed. PMID- 15815244 TI - Second Anglo-American Neuroanesthesia Conference May 21, 1988. PMID- 15815247 TI - Neuroanesthesiology: coming of age. PMID- 15815248 TI - Tolerance to N2O-induced alterations in somatosensory evoked potentials. AB - The effect of nitrous oxide (N2O) on somatosensory evoked potentials from the cortical (CEP) and spinal cord (SCP) regions in response to forepaw stimulation was studied in ketamine-anesthetized and mechanically ventilated rats. The CEP was recorded from the skull over the contralateral somatosensory area; the SCP was recorded from the supraspinous ligament at C57-6 and L1-2 levels of the spine. Rats were exposed to 70% N2O for 5 h, whereupon N2O was withdrawn for 2 h. Thereafter, the rats were re-exposed to N2O for 10 min. The N13-P21 component of the CEP, the slow positive wave (P2) of the segmental SCP, and the heterosegmental positive cord dorsum potential (HSP) were significantly suppressed by N2O, while the large negative (N1) component of the segmental SCP remained unchanged. A partial recovery of the CEP and HSP was observed during the 5 h of N2O anesthesia, while significant recovery of the P2 component of the SCP was not observed. The withdrawal from N2O following 5 h exposure caused an augmentation of the CEP (When compared to the control values). Re-exposure of rats to N2O again caused the suppression of these potentials as in the initial exposure. The results suggest that the phenomenon of tolerance to N2O in terms of evoked potentials develops within 5 h in the brain but not in the spinal cord. PMID- 15815249 TI - Effects of hypoxic hypoxia and reoxygenation on H2O2 production in rat brain in vivo. AB - The effects of hypoxic hypoxia and subsequent reoxygenation on hydrogen peroxide (H2O2) production was studied in the rat brain in vivo. Brain H2O2 production was measured by H2O2-dependent aminotriazole inactivation of endogenous brain catalase activity. Brain catalase activities of rats breathing air (0.2 ATA O2, control) were 168 +/- 5 (n = 10), 125 +/- 4 (n = 6), and 100 +/- 5 (n = 8) U/g brain (mean +/- SEM) at 0, 30, and 60 min after i.p. aminotriazole injection, respectively. Catalase activities after exposure to 5% O2 with N2 for 15 min, 10% O2 with N2 for 30 min, and 6% O2 with nitrous oxide (N2O) for 15 min were 131 +/- 4 (n = 7), 122 +/- 6 (n = 5), and 124 +/- 6 (n = 7) U/g brain, respectively, at 30 min after aminotriazole injection, and were not significantly different from each other or control. Reoxygenated on room air, 100% O2, and hyperbaric 3 ATA O2 for 30 min immediately after each period of hypoxia, brain catalase activity at 60 min after aminotriazole injection in the group of pre-exposure to 6% O2 with N2O was 67 +/- 3, 74 +/- 3, and 67 +/- 6 U/g brain with 0.2 ATA O2 (n = 6), 1.0 ATA O2 (n = 5), and 3.0 ATA O2 (n = 5), respectively. All of these were significantly different from control and other hypoxic pre-exposure groups with N2 (p <0.01) but not from each other. Reoxygenation of the brain after hypoxia with N2O could exacerbate cerebral damage by increasing oxygen free radical production. PMID- 15815250 TI - EEG changes during carotid occlusion and hypotension in the rat: the effects of nitrous oxide. AB - It has been suggested that N2O may alter the sensitivity of the brain to ischemia. To test this hypothesis. we examined the effects of N2O on the development of left-right hemispheric asymmetry in the electroencephalogram (EEG) during hemorrhagic hypotension in rats subjected to unilateral carotid occlusion. Rats were anesthetized with halothane/O2/air, and ventilated to normocarbia (PaCO2 approximately 40 mm Hg). Catheters were placed in the femoral artery and vein, and both common carotid arteries (CCA) were exposed. Bilateral fronto occipital screws were then placed to record left and right hemispheric EEGs, which were processed by computer and stored on disc. Animals were then randomly assigned to one of three treatment groups (n = 8 each): group 1, 0.5 MAC (0.5%) halothane + 0.5 MAC (70%) N2O; group 2, 1 MAC (1.0%) halothane + 70% nitrogen; and group 3, 1 MAC halothane + 70% N2O. After stabilization, the left CCA were occluded. Animals with EEG changes at this point were discarded. Beginning 5 min later, venous blood withdrawal was started at a rate of 0.25 ml/min, while mean arterial blood pressure (MAP) and EEG were continuously recorded. After exsanguination was complete, EEG data (raw and processed) were re-examined by an individual who was unaware of the anesthetics administered to determine the MAP at which any evidence of EEG asymmetry appeared. There were no intergroup differences in weight, PaO2, PaCO2, pHa, blood glucose. hematocrit, or starting (prebleed) MAP. The earliest change in the EEG was typically a decrease in total amplitude over the hemisphere ipsilateral to the carotid occlusion. Adding 70% N2O to a 1 MAC halothane background (group 2 vs. group 3) had no effect on the MAP at which this EEG asymmetry appeared (54 +/- 13 vs. 53 +/- 10 mm Hg). However, this MAP was significantly higher in animals breathing 0.5 MAC halothane + 0.5 MAC N2O (group 1, 78 +/- 17 mm Hg, p = 0.0019 by ANOVA). We conclude that 70% N2O had no direct effects on the MAP at which EEG abnormalities appear (group 2 vs. 3), and that the observed differences are more closely related to the concentration of volatile agent. Whether these differences are related to anesthetic-induced differences in the brain's tolerance to reduced CBF or whether there are differences in cerebral blood flow (CBF) autoregulation are unknown. PMID- 15815251 TI - Moyamoya disease: pathophysiology and anesthetic management. AB - Moyamoya disease, an ischemic cerebrovascular disease, is characterized by a slowly increasing bilateral occlusion of the internal carotid circulation. Although collateral pathways are formed, therapy is aimed at further increasing blood flow by surgical anastomosis before ischemic events and fixed neurologic defects occur. This disease remains one of the few indications for performance of the operation of superficial temporal artery to middle cerebral artery anastomosis. Anesthetic considerations involve increasing substrate supply and decreasing demand for its use. Two cases of moyamoya disease are described, noteworthy not only because of the rarity of the disease but because of its occurrence in the Hispanic race rather than the traditional appearance in those only of Japanese descent. The anesthetic management is outlined and the literature reviewed. PMID- 15815253 TI - The neuroendocrine axis. PMID- 15815254 TI - Effect of a 21-aminosteroid oxygen free radical scavenger on neurological outcome following complete cerebral ischemia in dogs. PMID- 15815255 TI - The 21-aminosteroid U74006F does not improve recovery from incomplete cerebral ischemia. PMID- 15815256 TI - Does phenylephrine-induced hypertension during focal cerebral ischemia aggravate brain edema? PMID- 15815257 TI - Changes in cerebral blood flow (CBF) and cerebral oxygen consumption (CMRO2) during N2O in unanesthetized unrestrained rats. PMID- 15815258 TI - Glucose administration prior to cardiac arrest worsens neurologic outcome in cats. PMID- 15815259 TI - Acute glycemic normalization in the chronically hyperglycemic diabetic rat produces a cerebral hypermetabolic state. PMID- 15815260 TI - Glucose utilization during alfentanil-induced seizures in rats. PMID- 15815261 TI - Moderate brain trauma and shock decrease cerebral blood flow and cerebral oxygen delivery. PMID- 15815262 TI - Studies of learning and memory in invertebrates. PMID- 15815263 TI - The effects of transient hyperglycemia on brain glucose in halothane-anesthetized rats. PMID- 15815264 TI - The "risk" of nitrous oxide in neurosurgical patients operated upon in the sitting position: a prospective, randomized study. PMID- 15815265 TI - Prevention of intracranial hypertension at intubation with preservation of cerebral perfusion pressure during induction: cerebral protective effect of etomidate-induced burst suppression on EEG. PMID- 15815266 TI - The transcranial Doppler can image microaggregates of intracranial air and particulate matter. PMID- 15815267 TI - Correlation of intravenous vs. intracarotid cerebral blood flow (CBF) values. PMID- 15815268 TI - Cerebrospinal fluid pressure in anesthetized patients with brain tumors: impact of fentanyl vs. alfentanil. PMID- 15815269 TI - Cerebrovascular responses to sufentanil citrate (SC) in primates with and without intracranial hypertension. PMID- 15815270 TI - Concentration-related changes in the rate of CSF formation and resistance to reabsorption of CSF during enflurane and isoflurane anesthesia in dogs. PMID- 15815271 TI - The influence of cardiopulmonary bypass on cerebral autoregulation and CO2 responsiveness in the early postoperative period. PMID- 15815272 TI - Anesthesia for surgery for pediatric brain tumors. PMID- 15815273 TI - Anesthesia for diagnostic procedures and nonsurgical treatment in children with brain tumors. PMID- 15815275 TI - Cerebral function monitoring during carotid endarterectomy. PMID- 15815276 TI - A comparison of computerized EEG with internal carotid artery stump pressure for detection of ischemia during carotid endarterectomy. AB - Previous investigations appear to indicate that an ischemic EEG is not observed during carotid cross-clamping when the stump pressure is >/=60 mm Hg. In this report of 124 carotid endarterectomies (CEA) performed with selective shunting based on computerized EEG (CEEG), we compared the CEEG and this previously established critical stump pressure level of 60 mm Hg as methods of detecting cerebral ischemia during carotid clamping. A significant association between stump pressure and CEEG findings during clamping existed (p <0.05). Only 1 of 44 patients with a stump pressure >/=60 mm Hg received a shunt based on CEEG signs of cerebral ischemia. However, 62 of the remaining 80 patients did not receive a shunt and awoke neurologically intact despite a stump pressure <60 mm Hg. A highly significant association between the postoperative neurologic exam and the CEEG findings during carotid clamping was demonstrated (p <0.001). In contrast, for stump pressure, a correlation with the neurologic exam was not found. Compared to CEEG, these results appear to indicate that a critical stump pressure of 60 mm Hg is a sensitive but not specific indicator for the placement of a shunt selectively during CEA. The combined use of these two monitors should lead to reliable shunt selection, especially when stump pressure is <60 mm Hg. PMID- 15815277 TI - Distribution of cerebral blood flow during deep isoflurane vs. pentobarbital anesthesia in rats with middle cerebral artery occlusion. AB - While previous studies have identified a protective effect for barbiturate anesthesia during focal cerebral ischemia, no such effect has been demonstrated for isoflurane. To better understand the effects of these anesthetics on cerebral blood flow and metabolism that might have relevance to their respective potential for cerebral protection, fasted physiologically stable rats underwent autoradiographic determination of CBF and CMRglu during deep isoflurane or pentobarbital anesthesia (burst suppression of EEG). As expected, cerebral blood flow was significantly greater during isoflurane anesthesia (isoflurane = 157 +/- 18 and pentobarbital = 54 +/- 12 ml/100 g/min) while CMRglu values were nearly identical (isoflurane = 35 +/- 5 and pentobarbital = 33 +/- 4 mumol/100 g/min). Additional identically anesthetized rats underwent middle cerebral artery occlusion with CBF autoradiographically determined 1 h later. While the insult resulted in a significant reduction in the ipsilateral hemispheric and cortical blood flow in both anesthetic groups, flow remained at least twofold greater in isoflurane as opposed to pentobarbital-anesthetized rats. When regional flow was assessed, no difference between groups was observed with respect to the area of tissue with flow values falling between 0-10 ml/100 g/min. In contrast, isoflurane-anesthetized rats had significantly less hemispheric and cortical area with flow values in the ranges of 10-20 and 20-30 ml/100 g/min, respectively. These data, therefore, do not support the contention that isoflurane causes maldistribution of CBF during focal ischemia. PMID- 15815278 TI - Effect of three pretreatment techniques on hemodynamic and CSFP responses to skull-pin head-holder application during thiopentone/isoflurane or propofol anesthesia. AB - The effects of three different pretreatment techniques on ICP and MAP responses to pin head-holder application (HH) were studied. Thirty-two patients for elective intracranial surgery were assigned randomly to three pre-treatment groups: group I-local scalp infiltration with lidocaine (6-8 ml of 2% solution); group II-deepening of general anesthesia with either thiopentone (2 mg/kg) or propofol (0.6 mg/kg); and group III-intravenous fentanyl (4 mug/kg). Two anesthetic techniques were used, subdividing the three groups into subgroup A (thiopentone-isoflurane) and subgroup B (propofol). Measurements of MAP, cerebrospinal fluid pressure (CSFP), cerebral perfusion pressure (CPP), and heart rate (HR) were made at pretreatment, just before HH (time T0: baseline), and 0.5, 1, 2, 3, 4, and 5 min after HH. Within each group, MAP, HR, CPP, and CSFP varied similarly in both subgroups, so the data from A and B were cumulated. After HH application, MAP, HR, CPP, and CSFP increased in all groups, but significantly less in group I. The maximal MAP increase occurred at T1 and was 6.7 +/- 2.2% in group I, 27.9 +/- 4.9% in group II, and 27.1 +/- 6.5% in group III (difference between I and II-III: p <0.005). HR increased similarly in the three groups, but less in group I. The maximal CSFP increase occurred at T0.5 and was 12.2 +/- 10.0% in group I, 31.9 +/- 10.8% in group II, and 24.5 +/- 5.6% in group III (difference between I and II-III: p < -0.05). The changes in CPP paralleled MAP changes. In conclusion, the thiopentone-isoflurane sequence and continuous propofol anesthesia provide similar hemodynamic conditions and responses to nociceptive stimuli; local lidocaine infiltration achieves better control of MAP, HR, CSFP, and CPP after HH application than the deepening of general anesthesia or intravenous fentanyl. PMID- 15815279 TI - External ventricular drainage and passive vs. active neurosurgical intervention in the management of hypertensive intracerebral hemorrhage with rupture into the ventricles. AB - The mortality rate of hypertensive intracerebral hemorrhages (HICH) with ventricular rupture (VR) has been said to range between 60 and 86%. We report on 33 consecutive cases of HICH associated with VR, which were diagnosed by CT and treated by external ventricular drainage (EVD). Sixteen hemorrhages were in the capsulolenticulostriate area, and 6 were in the posterior fossa. A modification of the usual system of severity grading for intraventricular hemorrhages (IVH) in posterior fossa bleedings is suggested. Extent of IVH correlated with neurological deficit on admission in 29 patients (87%) and with outcome in 26 patients (78%). Following passive neurosurgical policies in which the deterioration of patient's clinical situation indicates the craniotomy, only one intracerebral hematoma (ICH) was partially removed. Twelve patients (36.5%) died, and 12 of the survivors had no or only a minor neurological deficit. The results are compared with those of a series of patients, reported by Hayashi et al., who were treated according to active neurosurgical policies in which the largeness of ICH and IVH indicates the craniotomy, whereby it is concluded that active neurosurgical intervention does not reduce significantly the mortality and morbidity of HICH with VR. The pathophysiological explanation for this is discussed on the basis of a review of reference literature. PMID- 15815280 TI - Flumazenil does not impair autoregulation of CBF in dogs when given with or without prior administration of midazolam. AB - The effects of flumazenil (a benzodiazepine antagonist) on autoregulation of cerebral blood flow (CBF) were examined in dogs receiving midazolam and in dogs not receiving midazolam. Both groups were anesthetized with halothane (0.3% end expired) and nitrous oxide (66%) in oxygen. Auto-regulation of CBF was assessed by determining the slope relating CBF to cerebral perfusion pressure (CPP). These CBF/CPP slopes were derived from measurements of CBF at each of three CPPs: 65, 80, and 95 mm Hg. The effects of flumazenil on autoregulation were examined by comparing CBF/CPP slopes prior to administration of flumazenil to CBF/CPP slopes after flumazenil doses of 0.0025 and 0.16 mg/kg, both when cerebrospinal fluid (CSF) pressure was normal and when CSF pressure was elevated to 30 mm Hg. Prior to administration of flumazenil, CBF/CPP slopes were not different from zero, indicating intact autoregulation of CBF. CBF/CPP slopes did not change following administration of flumazenil, indicating that autoregulation of CBF was not substantially impaired. In dogs with normal CSF pressure that were receiving midazolam, both doses of flumazenil altered the electroencephalogram and the highest dose of flumazenil decreased cerebral vascular resistance and increased CBF and CSF pressure. No such changes were seen at the other experimental conditions. It is concluded that flumazenil does not severely disturb autoregulation of CBF, although flumazenil 0.16 mg/kg causes a statistically significant increase of CBF in dogs with normal CSF pressure that are receiving midazolam. PMID- 15815281 TI - Regional brain glucose utilization in the conscious rat exposed to prolonged hypobaric hypoxia. AB - Because of the physiologic and metabolic changes that occur during acclimatization, we hypothesized that LCGU may be normal during prolonged hypoxia. We exposed five Sprague-Dawley rats to hypoxia (air at 380 mm Hg) for 2 weeks (hypoxic group) and six rats to 2 weeks of hypoxia followed by 2 weeks of recovery in room air (recovered group). Six control rats breathed room air (control group). Regional brain glucose utilization was measured in awake animals by using 2-[C]deoxyglucose autoradiography. Glucose utilization was comparable in the control and recovered groups and in most brain regions of hypoxic animals. Glucose utilization was decreased slightly in 10 of 12 gray matter regions examined and was 20 to 25% lower (p <0.05) in the olfactory and auditory cortices, the caudate nucleus, and the superior olive of the hypoxic group. White matter glucose utilization was unchanged. Hypoxic rats, compared to controls, had a lower PaO2 (53 +/- 3 vs. 76 +/- 3 mm Hg, mean SEM, respectively), a lower PaCO2 (22 +/- 1 vs. 36 +/- 2 mm Hg), and a higher mean pulmonary artery pressure (46 +/ 3 vs. 14 +/- 3 mm Hg) and hematocrit (61 +/- 2% vs. 48 +/- 1%; p <0.005 for all comparisons). Pulmonary hypertension and polycythemia persisted in recovered rats. Arterial pressure, pH, and plasma glucose were unaffected. Therefore, while acute hypoxia may increase glucose utilization in most brain structures, prolonged exposure does not. PMID- 15815282 TI - Concentration-related changes in the rate of CSF formation and resistance to reabsorption of CSF during enflurane and isoflurane anesthesia in dogs receiving nitrous oxide. AB - The rate of cerebrospinal fluid (CSF) formation (Vf) and resistance to reabsorption of CSF (Ra) were determined at four concentrations of enflurane (0.9, 1.8, 2.0, and 3.5% end-expired) or isoflurane (0.6, 1.1, 1.7, and 2.2% end expired) in dogs receiving nitrous oxide (66%) in oxygen. At the two higher concentrations of enflurane, Vf was about 27% greater (38-40% greater when corrected for the effects of time) than at the two lower concentrations of enflurane. At the two lower concentrations of enflurane and at all doses of isoflurane, Vf did not differ between or within anesthetic groups (0.029-0.033 ml/min). The pattern of Ra data was more varied. Ra was elevated with the two lower concentrations of enflurane (by about 68%) and with isoflurane 1.1% (by about 33%), and reduced with the two higher concentrations of isoflurane (by about 14-17%). It is concluded that the effects of enflurane and isoflurane on CSF dynamics are concentration-related. All concentrations of enflurane studied favor CSF volume expansion, while high concentrations of isoflurane favor CSF volume contraction. PMID- 15815284 TI - Brain metabolism during ischemia and anoxia. PMID- 15815285 TI - Mechanisms of ischemic cerebral damage: the role of glial cells. AB - Glial cells exert an important homeostatic function in the brain in protecting neurons from extracellular mediators of secondary brain damage. In vitro studies suggest that glial swelling as observed in ischemia is not simply an indication of glial damage but rather of the activation of glial homeostatic functions. The mechanisms of glial swelling due to exposure to glutamate and acidosis, as well as therapeutic consequences, are discussed. PMID- 15815286 TI - Benzodiazepines and antagonists: effects on ischemia. PMID- 15815287 TI - Barbiturates for global and focal ischemia. PMID- 15815289 TI - Volatile anesthetics and the ischemic brain. PMID- 15815288 TI - Propofol: effects on indices of cerebral ischemia. AB - The effects of propofol given before and during a period of profound hypotension that caused incomplete global cerebral ischaemia were investigated in anaesthetised cats. Cortical cerebral blood flow, extracellular fluid pH, potassium and calcium ion activities, and electroencephalogram were recorded. Neuropathological outcome was also assessed. Propofol-treated animals had higher cerebral blood flows than control animals after the period of hypotension (p <0.05); they also had better correction of extracellular fluid acidosis and hyperkalaemia (p <0.01) and a late improvement in calcium ion activity (p <0.05). Neuropathological outcome was not significantly different between the groups. PMID- 15815291 TI - The control of quality. PMID- 15815292 TI - Current trends in neurosurgical critical care medicine. AB - We conducted a survey of neurosurgeons to discover how they manage specific problems commonly seen in critically ill neurosurgery patients and compared the responses with relevant investigations published in the neurosurgical literature over the past 10 years. The 101 Chiefs of Neurosurgery of every North American academic training program were sent copies of two consecutive surveys for distribution to their attending staff. Among the topics covered were indications for steroid administration, prophylaxis of gastrointestinal bleeding, indications for insertion of ICP monitoring devices and use of prophylactic antibiotics while they are in place, and fluid restriction in patients with cerebral edema. Replies were strictly anonymous. A Medline Computer search for the period 1976-1989 was carried out for each topic. We received 136 and 67 replies each to our two surveys. Over 90 clinical and laboratory studies published in major medical journals, many randomized and prospective, were reviewed. Two trends were evident: (a) a noticeable lack of uniformity among neurosurgeons with respect to management of the same problem; (b) little or no basis of support in the literature for many management practices. Based on these studies and the responses to our survey, we suggest an examination of current clinical practices. PMID- 15815293 TI - Nitrous oxide decreases substance P receptor binding in the rat spinal cord. AB - Synaptic transmission of substance P (SP) in the spinal cord during nitrous oxide (N2O) exposure was examined in rats. In the analgesia study, 75% N2O significantly increased tail-flick latency by 57% during the first 2-h N2O exposure and by 19% after a second 2-h N2O exposure, suggesting development of acute tolerance. In the SP content study, SP in the dorsal part of the spinal cord significantly increased by 120% at the end of first N2O exposure, but returned to the control level by the end of second N2O exposure. In the receptor binding study, N2O significantly decreased the [I]Bolton-Hunter-SP ([I]BH-SP) binding (SP-P) in laminae I-II of the dorsal horn by 19, 18, and 18% at the end of first N2O exposure, at the end of the second N2O exposure, and at 4 h after the second N2O exposure, respectively, as compared to the control. This decrease in the [I]BH-SP binding became statistically insignificant by 12 h after the second N2O exposure. In contrast, [I]Bolton-Hunter-eledoisin binding (SP-E) remained unchanged. In the equilibrium binding study, the number of SP-P receptor (Bmax) in laminae I-II of the dorsal horn significantly decreased by 24, 38, and 30% at the end of the first and at the end of the second N2O exposure, and at 4 h after the second N2O exposure, respectively, without significant changes in the affinity (Ka). The present study demonstrated that tolerance to N2O analgesia occurred during the two repeated N2O exposures. SP receptor binding decreased and remained decreased 4 h after discontinuation of N2O. This evidence is the first demonstration of the functional changes caused by N2O in SP synaptic transmission of the spinal cord. PMID- 15815294 TI - The use of transcranial Doppler sonography in the diagnosis of brain death. AB - The diagnosis of brain death is important for many reasons. Clinical testing may not be feasible, so that cerebral angiography is needed to confirm circulatory arrest. Angiography is, however, cumbersome and expensive. We present the results of transcranial Doppler (TCD) studies on the middle cerebral arteries of 40 patients with brain death. In six, no cranial signals could be obtained. All except two of the remaining patients had typical TCD appearances, with a reverberating pattern and little or no net forward flow. One patient had this appearance on one side and a carotid-cavernous fistula on the other side, and another had preserved middle cerebral artery flow. Sixteen control patients all had quite different signals, with some showing evidence of raised intracranial pressure and some of vasospasm. There were thus few false-negative results, and more importantly no false-positives. Refinement of this technique, especially looking to a numerical value for the net flow velocity below which circulatory arrest is certain, is needed. The possibility of a changing pattern on serial studies, with prediction of brain death before it actually occurs, is also to be explored. PMID- 15815295 TI - Alfentanil infusion in neurosurgical patients. AB - Alfentanil administered by constant rate infusion was evaluated as a main anesthetic in craniotomy patients. Eleven patients (ASA II) received either an induction dose of 200 microg/kg alfentanil followed by a 1 microg/kg/min infusion, or a 150 microg/kg bolus followed by a 1.5 microg/kg/min infusion. The infusion was stopped at least 1 h before the end of surgery. In addition to routine clinical neuroanesthesia monitoring, cardiovascular parameters, extubation conditions, and recovery scores were systematically recorded. Plasma alfentanil levels were determined in all patients during and for 6 h after the infusion. Finally, pharmacokinetic studies were carried out in five patients from both groups. This technique provided a remarkable cardiovascular stability, excellent extubation conditions, a comfortable recovery period, and no signs of prolonged or recurrent narcotization. No difference in clinical signs or alfentanil blood concentrations between the two drug regimens could be found. Pharmacokinetic parameters found were comparable to those obtained after single bolus drug elimination. Despite large individual variations and sometimes very prolonged infusion, there was no evidence of delayed drug administration. All of the above findings warrant the use of alfentanil infusion in neuroanesthesia and confirm the unique pharmacological properties of this drug among existing narcotics. PMID- 15815296 TI - Modulation of cerebrospinal metabolic responses to peripheral stimulation by enflurane anesthesia in rats. AB - Enflurane-induced modulation of cerebrospinal metabolic responses to peripheral nerve stimulation was examined in 30 rats. Local glucose utilization in the brain and lumbar spinal cord was measured using the autoradiographic 2-[C]deoxyglucose method at three anesthetic concentrations (0,5, 2, and 4%) either with or without electrical stimulation (5 mA, 0.5 ms, 10 Hz) of the unilateral sciatic nerve. Stimulation produced a 71 to 111% increase in glucose utilization in the ipsilateral dorsal horn of the spinal cord at all anesthetic concentrations examined. Stimulation also produced a 32 to 48% increase in glucose utilization in the hindlimb projectionarea of the contralateral somatosensory cortex at the two lowest concentrations (0.5 and 2%), while at 4% no stimulus-induced increase in glucose utilization was observed. The results show that there is a threshold at which enflurane suppresses the metabolic responses to peripheral stimulation in the somatosensory cortex but not in the spinal cord. If electrical stimulation of a peripheral nerve is regarded as analogous to surgical stimulation, considerable increase in the spinal cord metabolism may occur during surgery even in a deeply anesthetized subject. PMID- 15815297 TI - Loss of somatal neuropeptide y immunoreactivity in the rat hippocampus following transient cerebral ischemia. AB - Adult male Wistar rats were subjected to 20 min of global cerebral ischemia and allowed to survive for 1, 2, 4, or 21 days. The brains were processed for immunocytochemistry and the hippocampal neuropeptide Y (NPY)-immunoreactive (-i) neurons were counted and compared to control values. In order to map out the subregional distribution of ischemic cell loss in the hippocampus, cells were also counted in hematoxylin-eosin (HE)-stained brain sections processed from additional ischemic rats after 21 days survival. Cell counts demonstrated a significant loss of hippocampal NPY-i somata 1-21 days after ischemia. The ischemic loss of somatal NPY-i was in the CAI stratum oriens, the CA1 stratum radiatum, and the CA3(ab) subfield not correlated to hippocampal cell loss. NPY-i fibers were found in all subfields of the hippocampus 1-21 days after ischemia. It is known that the majority (>50%) of hippocampal somatostatin-i (SS) neurons also costore NPY-i and the SS-i neurons in the CA1 and CA3(ab) regions of the hippocampus are preserved following an ischemic insult. The present results showed a 90% ischemic loss of CA1 and CA3 NPY-i somata. Based on these findings, it is concluded that ischemia selectively damaged NPY-i and not SS-i within some surviving hippocampal neurons that co-localized both peptides prior to the ischemia. PMID- 15815298 TI - Naloxone: report of a beneficial side effect. AB - We report two cases. The first was an unexpected, complete resolution of an acute unilateral neurologic deficit associated with anesthesia when naloxone was administered to reverse residual narcotic effect. The second was a complete resolution of postoperative unilateral electroencephalographic evidence of ischemia after naloxone administration in a patient following a carotid endarterectomy. A literature search suggests that naloxone may be useful in the treatment of acute and hyperacute stroke due to ischemia. PMID- 15815300 TI - Ion distributions in brain during ischemia. AB - The function of the central nervous system-and other organs-depends upon preservation of ionic gradients across cell membranes. In nervous tissue, the ion gradients are especially important since generation of action potentials and synaptic processes relies on transfer of ions across the plasma membrane. This report describes the fact that anoxia profoundly changes the brain interstitial ion milieu. Impaired ATP regeneration starts the chain of events that cause a breakdown of ion homeostasis. The pronounced ionic changes are not caused by impaired ion pumping but rather by opening of ion channels-probably most importantly via release of transmitter substances. Despite the severity of the ionic changes, the brain interstitial ion environment is readily normalized after the anoxic episode. The ionic disturbances are not the cause of the functional deficits encountered in anoxia but are probably of significance in the irreversible neuronal damage evolving after anoxia. PMID- 15815301 TI - NMDA antagonists and hypoxic neuronal injury. PMID- 15815303 TI - Neuronal calcium channels. PMID- 15815302 TI - Magnesium and the injured brain. PMID- 15815304 TI - Effects of Ca2+-entry blockers on ischemic brain. PMID- 15815305 TI - Responsiveness of the cerebral circulation to acute alterations in mean arterial pressure during the administration of propofol. AB - The responsiveness of the cerebral circulation to acute increases in mean arterial pressure was studied before and during the administration of propofol 3, 6, or 12 mg/kg/h in the anaesthetised baboon. Although mean arterial pressure increased significantly on each occasion, there were no significant changes in cerebral blood flow. This indicates that the physiological responsiveness of the cerebral circulation to alterations in mean arterial pressure was preserved during the administration of propofol in the concentrations studied. PMID- 15815306 TI - Neuroplasticity and repair following injury to the central nervous system. PMID- 15815307 TI - Local anesthetics in cerebral ischemia. AB - Local anesthetics are thought to be potentially beneficial in ischemia because they reduce cerebral energy expenditure and may prevent ischemic membrane depolarization. In vitro studies and in vivo studies in models of focal ischemia have indeed shown a benefit from the prophylactic use of lidocaine. Results in global ischemia have been conflicting. The effects of lidocaine may reflect a neurovascular rather than a membrane-stabilizing effect. PMID- 15815309 TI - Muscle relaxants in neurosurgical anesthesia: what should we do when the pressure's on? PMID- 15815310 TI - Succinylcholine-induced increases in CSF pressure are not affected by PaCO2 or mean arterial pressure in dogs. AB - The effects of PaCO2 and mean arterial pressure (MAP) on succinylcholine-induced changes in cerebrospinal fluid (CSF) pressure were examined in 31 dogs anesthetized with halothane (0.7% end-expired) and nitrous oxide (66% inspired) in oxygen. Group 1 (n = 9) received succinylcholine (1.5 mg/kg i.v.) without control of PaCO2 or MAP. Group 2 (n = 12) received succinylcholine with CO2 maintained at control, and no regulation of MAP. Group 3 (n = 10) received succinylcholine without regulation of PaCO2 and maintenance of MAP at control values with trimethaphan. Succinylcholine produced an increase in CSF pressure of 1.7 +/- 1.7-3.9 +/- 3.6 cm H2O (mean +/- SD). There were no differences in CSF pressure increase between groups. The increase in CSF pressure was compared to the variation in the uncontrolled physiological parameter (i.e., CSF pressure was compared to PaCO2 and MAP in group 1, MAP in group 2, and PaCO2 in group 3). The CSF pressure increase over time did not strictly parallel changes in PaCO2 and/or MAP, and the magnitude of increase of CSF pressure did not correlate significantly with PaCO2 and/or MAP. It is concluded that, in anesthetized dogs, succinylcholine produces relatively small increases in CSF pressure whether or not PaCO2 or MAP is controlled. These results do not support the hypothesis that a succinylcholine-induced increase in CSF pressure is due chiefly to an increase in PaCO2 and/or MAP and can be abolished by preventing those changes. PMID- 15815311 TI - Cerebrovasculr response to CO2 in edematous brain during either fentanyl or isoflurane anesthesia. AB - Cerebral blood flow response to changes in PaCO2 was studied in the edematous cerebral cortex of 19 patients with malignant supratentorial tumors using laser Doppler flowmetry technology. General anesthesia for craniotomy was induced with thiopental, 3-5 mg/kg i.v., and N2O, 60% in O2. In random sequence, 8 patients were assigned to receive fentanyl, 6 +/- 1.6 (SEM). mug/kg i.v.; the other 11 received isoflurane, 0.56% end-tidal + 0.07 (SEM). After a craniotomy bone flap was turned and the dura was opened, laser flowmetry probes were placed over surgically undisturbed cortex that was known to be edematous from preoperative CT and MRI scans. Flow index measurements were first made at hypocarbia (PaCO2 = 24.2 +/- 0.9 and 21.5 +/- 2.1 mm Hg for the fentanyl and isoflurane groups, respectively). Minute ventilation was then decreased and cortical flow index was remeasured with PaCO2 = 34.2 +/- 0.6 and 33.0 +/- 0.8 mm Hg for the fentanyl and isoflurane groups, respectively. Hypocarbia during fentanyl-supplemented N2O-O2 anesthesia resulted in a cortical flow index that was 70 +/- 8% of the flow index at near normocarbia (p <0.05). During isoflurane N2O-O2 anesthesia, however, there was a wide variety of responses to hypocarbia, including three patients whose flow indices increased markedly. The mean flow index during hypocarbia was significantly (p <0.05) lower during fentanyl-N2O anesthesia than it was during isoflurane-N2O anesthesia. There was no predictable relationship between the type of brain tumor and the CBF response to hypocapnia during isoflurane-N2O anesthesia. It is concluded that, in edematous brain, cerebral cortical blood flow response to hypocarbia is more likely to be preserved during fentanyl supplemented N2O-O2 anesthesia than it is during isoflurane-supplemented N2O-O2 anesthesia. In neuropathologic states where hyperventilation is thought to be necessary to reduce cerebral blood flow and decrease brain bulk, isoflurane may be less satisfactory than fentanyl as a supplement to N2O-O2 anesthesia. PMID- 15815312 TI - Perioperative monitoring of the electrocardiogram during cerebral aneurysm surgery. AB - Electrocardiographic (ECG) abnormalities occur frequently following a subarachnoid hemorrhage and may also occur intraoperatively and postoperatively in patients undergoing neurovascular procedures. The aim of this study was to assess the relationship between ECG changes and the neurological status of the patient, the size and the location of the aneurysm, and the influence of these changes on the cardiac and neurological outcome. The preoperative ECG was analyzed in 270 patients. Forty-five patients had intraoperative Holter monitoring. An immediate postoperative ECG was recorded in 120 patients and 60 patients had three consecutive postoperative ECGs. Preoperatively, 52% of the patients had an abnormal ECG and the incidence was highest in patients with a poorer neurological status. Most of the ECG changes involved the T wave or the ST segment. Intraoperative and postoperative changes occurred in 35 and 65% of the patients, respectively, and were independent of the studied factors. There were no documented cardiac events. The presence of an abnormal preoperative ECG did not influence the neurological outcome of the patient, but fluctuating postoperative changes were associated with a worse outcome. PMID- 15815313 TI - The equivalence of anesthetic regimens with respect to plasma glucose elevation during cerebrovascular surgery. AB - Hyperglycemia, even if mild, is known to aggravate neuronal damage from cerebral ischemia. In order better to define the influence of currently used anesthetic techniques on plasma glucose levels during cerebrovascular surgery, we examined serial plasma glucose values during 43 carotid endarterectomies (CEA) and 19 intracranial arteriovenous malformation (AVM) resections. CEA patients (aged 67.6 +/- 1.4 years and weighing 76.4 +/- 2.3 kg, mean +/- SEM) received N2O in O2 and either isoflurane (ISO) (n = 14), halothane (n = 8), fentanyl (n = 10), or sufentanil (n = 11). Plasma glucose was compared at 1.12 +/- 0.05 h (stage 1), 2.08 +/- 0.07 h (stage 2), and 3.12 +/- 0.1 h (stage 3) after induction of anesthesia. AVM patients received ISO and N2O in O2. Plasma glucose was compared 2.32 +/- 0.14 h (stage 1) and 6.25 +/- 0.34 h (stage 2) after induction of anesthesia (surgical stage). Glucose was determined by the hexokinase method. In the CEA cases, progressively elevated plasma glucose levels were associated with successive surgical stage (114 +/- 6, 122 +/- 6, and 138 +/- 6 mg/dl). The seven CEA patients that carried the diagnosis of diabetes mellitus tended to have higher glucose levels but they did not differ significantly from nondiabetic patients. The AVM patients (aged 35.7 +/- 2.3 years and weighing 71.1 +/- 2.9 kg) were all nondiabetic. They were significantly younger than the CEA patients and each received dexamethasone intraoperatively. In these patients, there was a significant effect (p <0.04) of surgical stage to increase plasma glucose (115 +/ 10 vs. 126 +/- 10 mg/dl). For CEA, the anesthetic techniques examined do not differ significantly in their influence on plasma glucose levels, but all techniques were associated with a gradual increase in plasma glucose levels intraoperatively, even in nondiabetic patients. Compared to the group of younger AVM patients, glucose elevation was more pronounced in the elderly CEA patients. We conclude that intraoperative monitoring of plasma glucose may be useful in elderly patients during prolonged neurovascular procedures. PMID- 15815314 TI - Pressure-supported ventilation for posterior fossa operation. AB - To maintain enough gas exchange while using spontaneous respiration as a monitor of the normal brainstem function, we tried pressure-supported ventilation (PSV) with a Servo 900C ventilator (Siemens Elema AB, Sweden) on 12 otherwise healthy patients during posterior fossa operation. Ventilation mode was switched from controlled to PSV after the dura was open uneventfully in all cases but one. With a trigger level of -1 to -2 cm H2O, spontaneous respiration was triggered to start the inspiration. With supporting inspiratory pressure of 4-20 cm H2O, PaCO2 was kept at 31.7-45.9 mm Hg. The ventilatory level could be monitored breath by breath by ventilatory frequency, tidal volume, minute volume, and end-tidal CO2 concentration shown on the ventilator system. Apnea was observed in two cases during surgical manipulation around the brainstem. It was indicated immediately by the ventilator's alarm for decreased expiratory minute volume, and no sign of brainstem dysfunction was observed postoperatively. PSV was useful in maintaining adequate ventilation whereas spontaneous respiration was used as an indicator of normal brainstem function. The alarm system of the ventilator was sensitive enough to detect the surgical invasion of the brainstem at a very early stage. PMID- 15815315 TI - Concurrent monitoring of brainstem auditory and somatosensory evoked potentials during carotid endarterectomy. AB - Intraoperative management of a 68-year-old man with a 3-month history of presyncopal attacks presenting for left carotid endarterectomy is reported. Preoperative angiograms revealed that the patient had a 90% stenosis of his left internal carotid artery and a 35% stenosis of right internal carotid artery, absent posterior communicating arteries, a totally occluded left vertebral artery, and a right vertebral artery that became compressed upon turning his head to the right, often precipitating the presyncopal attacks. The surgical procedure was managed with intraoperative monitoring of both brainstem auditory and somatosensory evoked potentials. The patient had an uneventful outcome. The anesthetic management is discussed. PMID- 15815316 TI - Systemic hypovolemia after subarachnoid hemorrhage. AB - Two patients who had suffered subrachnoid hemorrhage were subjected to intensive monitoring of parameters of circulating blood flow, cardiovascular dynamics, and fluid and electrolyte balance. Among the parameters studied were red cell volume, plasma volume, and total blood volume, cardiac output, and central venous and pulmonary capillary wedge pressures. Both patients experienced acute neurological deterioration during the study. Red cell volume fell in a premonitory fashion prior to neurological deterioration in both, and in one patient plasma volume and total blood volume decreased by almost 50% in the 5 days prior to neurological deterioration. Central venous and pulmonary capillary wedge pressures fell initially but did not predict the deterioration. Systemic vascular resistance fell prior to clinical deterioration in one patient following ventriculoperitoneal shunting, and in the other following craniotomy and subarachnoid drainage. The falls in systemic vascular resistance may have caused shunting of cardiac output away from the compromised cerebral circulation, thereby triggering neurological deterioration. PMID- 15815317 TI - Does sufentanil exacerbate marginal neurological dysfunction? PMID- 15815319 TI - Plasma concentrations of phenytoin after phenytoin loading during cardiopulmonary bypasss in adult patients undergoing pulmonary thromboendarterectomy with profound hypothermia and total circulatory arrest. PMID- 15815318 TI - Morbidity and mortality associated with rapid control of systemic hypertension in patients with intracranial hemorrhages. PMID- 15815320 TI - Argininosuccinate synthetase deficiency and reye syndrome-like presentation. PMID- 15815323 TI - Motor tract evoked potentials: is there a future in intraoperative monitoring? PMID- 15815324 TI - Suppression of motor evoked potentials by inhalation anesthetics. AB - The purpose of this study was to record evoked action potentials from forearm muscles in response to single-shock supramaximal electrical stimulation of motor cortex in room air and under different concentrations (0.5-1.5%) of isoflurane, enflurane, and halothane anesthesia in rats. Anesthesia was induced with a mixture of fentanyl and droperiodol, which was then followed by 10-min inhalation of each gas anesthetic under controlled ventilation. Increasing concentrations of isoflurane (n = 12) caused a progressive increase in onset latency and a decrease in peak-to-peak amplitude and duration. Similar increases in latency and decreases in amplitude and duration occurred under enflurane (n = 10) and halothane (n = 10) anesthesia. The three anesthetics caused a significant latency increase over baseline (room air) values for concentrations from 0.5 to 1.5% (p < 0.01). The amplitude and duration of muscle responses under all three volatile anesthetics at 0.5-1.5% concentrations were significantly lower than baseline (p < 0.01). Isoflurane, enflurane, and halothane anesthesia significantly altered the muscle response evoked by motor cortex stimulation in experimental animals. PMID- 15815325 TI - Effects of incremental ketamine hydrochloride doses on motor evoked potentials (MEPs) following transcranial magnetic stimulation: a primate study. AB - the cumulative dose effect of ketamine hydrochloride (KH) on transcranial magnetic-induced motor evoked potentials (MEPs) was examined in monkeys. Electromyographic (EMG) responses were recorded from the contralateral abductor pollicis brevis (APB) and abductor hallucis (AH) muscles. MEP brain stimulation threshold, latency, and amplitude values were studied. After obtaining baseline recordings, increments of KH (5 mg/kg every 15-20 min) were given i.v. (50 mg/kg total dose). MEPs were repeatedly recorded following KH injections. No loss of potentials was encountered in any animal. However, KH induced significant MEP latency delay in doses >/=35-40 mg/kg and amplitude depression in doses >/=15-20 mg/kg (p < 0.01). Under various KH doses, the amplitude depression ranged from 13.6 to 45.5% for APB and 57.3 to 82% for AH compared to the control values. The MEP latency prolongation ranged from 3.5 to 18% for APB and 4.2 to 13.1% for AH. The stimulation threshold rise ranged from 6.7 to 14.7% for APB and 7 to 17.9% for AH. Statistical correlation was closest between cumulative KH doses and MEP latency prolongation. We conclude that, in the primate model, reliable MEP recording is feasible under deep KH anesthesia. However, awareness of drug induced response alterations is essential during interpretation of intraoperative MEP changes. Further investigation is warranted regarding the specific dose effect in humans and safety of magnetic stimulation. PMID- 15815326 TI - Dose-dependent blood flow velocity changes in the Basal cerebral arteries following low-dose ketamine. AB - The effects of low-dose ketamine on blood flow velocity and pulsatility index (PI) in the basal cerebral arteries were studied in two groups of healthy volunteers (group A: 0.25 mg/kg, n = 10; group B: 0.5 mg/kg, n = 10) by means of a 2 MHz pulsed transcranial Doppler ultrasonic system (TCD) for an observation period of 70 min. In addition, mean arterial pressure (MAP), heart rate (HR), end tidal CO2 (petCO2), and oxygen saturation (saO2) were recorded. The mean blood flow velocity (Vmean) increased significantly by 28% in group A and by 68% in group B within 2-9 min. In group B, the PI was reduced by 32% for a period of 2-7 min (p < 0.05). MAP (group A: -20%; group B: -26%) and HR (group A: -39%; group B: -54%) increased significantly (p < 0.05). Vmean and PI were significantly different between groups A and B, suggesting a dose-dependent stimulation of cerebral hemodynamics. The flow velocity increases cannot be entirely explained by systemic hemodynamic changes since there was no intergroup difference with respect to MAP and HR. No significant differences were observed with regard to petCO2 and saO2. With the assumption that the diameter of the insonated vessel is not changed by ketamine, the dose-dependent increase in Vmean with a concomitant decrease in PI may be interpreted as indicating an increase in cerebral blood flow. PMID- 15815327 TI - Arterial CO2 tension and cerebral vascular reactivity during the induction of acute hypertension and hypotension in the awake human. AB - The CO2 reactivity was calculated at mean arterial blood pressure (MABP) values ranging from 40 to 140 mm Hg in 15 normotensive volunteers and in 7 patients with chronic arterial hypertension. The cerebral vascular reactivity (CVR) was estimated from indirect measurements of cerebral blood flow (CBF) by the arteriovenous oxygen saturation method. In all subjects but one of the hypertensive patients, there was a significant linear correlation of CVR with PaCO2 over the total range of MABP. The median CO2 reactivity in the volunteers and in the hypertensive patients was 2.7 and 2.8% CBF/0.1 kPa, respectively (NS). At MABP values corresponding to the plateau of CBF autoregulation (baseline MABP +/- 20%), the median CO2 reactivity was 3.0% CBF/0.1 kPa in both groups. During a cross-sectional correlation analysis of the pooled data from all volunteers, the CO2 reactivity increased with increasing MABP in the range of 50-110 mm Hg. Outside this range there was no correlation of CVR with PaCO2. In conclusion, our results confirmed that during acute hypo- and hypertension, the influence of PaCO2 on the CVR is reduced. The full CO2 reactivity of the cerebral vessel exists only at the plateau of CBF autoregulation. PMID- 15815328 TI - Monitoring of somatosensory evoked potentials during temporary arterial occlusion in cerebral aneurysm surgery. AB - Somatosensory evoked potentials (SSEP) have been used during cerebral aneurysm surgery to monitor the integrity of neural pathways. The purpose of this study was to evaluate the effectiveness of SSEP monitoring as a predictor of neurological outcome during temporary arterial occlusion. In a series of 157 patients monitored, 97 patients had temporary occlusion of the feeding artery. Twenty-three patients developed a SSEP change during temporary occlusion, 15 reversible (recovery of the change after the release of occlusion), and 8 persistent (no recovery) changes. A persistent change predicted a postoperative neurological deficit in each case, whereas, of the 15 patients with reversible changes, only 5 had postoperative deficits. Seventy-four patients had no change on SSEP monitoring but 10 patients did have new neurological deficits postoperatively. The false positive rate was 43% and the false negative rate was 14%. SSEP was a better predictor of neurological deficits in patients with aneurysms of the carotid circulation than of the vertebral-basilar arteries. Despite these limitations, we find SSEP monitoring useful during temporary occlusion in cerebral aneurysm surgery. PMID- 15815329 TI - EEG suppression and increased blood-brain barrier permeability following intracarotid injection of iothalamate meglumine (Conray) in dogs. AB - Over a 2-year period we observed three cases of unilateral suppression of the electroencephalogram (EEG) lasting from 45 s to 4 min following intracarotid injection of 60% iothalamate meglumine (Conray) for intraoperative carotid angiography postendarterectomy. As a result of these cases we undertook studies in 11 dogs anesthetized with isoflurane to examine causes of EEG suppression following intracarotid contrast medium injection. In group 1 (n = 6) cerebral blood flow (CBF), the cerebral metabolic rate for oxygen (CMRO2), EEG activity, and permeability of the blood-brain barrier (BBB) were determined. In group 2 (n = 5) cerebrospinal fluid (CSF) pressure, EEG activity, and BBB permeability were determined. Intracarotid injection of 5 ml of 60% Conray was associated with unilateral EEG suppression and increased BBB permeability in 1 of 11 dogs. Injection of contrast material caused no change in CBF or CMRO2 and caused a statistically significant but physiologically unimportant increase of CSF pressure (from 12 +/- 1 to 16 +/- 1 cm H2O, mean +/- SEM). It is concluded that EEG suppression following intracarotid injection of Conray is a rare event. It seems unlikely that EEG suppression resulted from cerebral ischemia or hypoxia, but rather was associated with increased BBB permeability. Increased BBB permeability likely was caused by the osmotic effect of Conray and not by hypoxic ischemic microvascular injury or loss of autoregulation of CBF. PMID- 15815330 TI - Effect of isoflurane-induced hypotension on cerebral autoregulation in the anesthetized pig. AB - The influence of isoflurane-induced hypotension on regional cerebral blood flow (rCBF) and cerebrovascular autoregulation was evaluated in 11 normocapnic pigs during anesthesia comprising fentanyl and nitrous oxide in oxygen. rCBF was determined as sagittal sinus outflow and recorded continuously by an electromagnetic technique. Regional cerebral metabolic rate for oxygen (rCMRO2) was calculated as rCBF multiplied by the arteriosagittal sinus oxygen content difference. Cerebral autoregulation was evaluated by two formal tests; blood pressure increase by infusion of angiotensin and blood pressure decrease by caval block. Light hypotension [mean arterial blood pressure (MABP) 94 +/- 3 mm Hg] and moderate hypotension (MABP 56 +/- 1 mm Hg) were achieved with the inspired concentrations of 1.7 +/- 0.2 and 2.7 +/- 0.2% isoflurane, respectively. rCBF was measured and the tests were performed before, during, and after isoflurane administration. Isoflurane produced no significant change in rCBF at either level of hypotension. At moderate hypotension the rCMRO2 was decreased by 40 +/- 6%. At both levels of hypotension, isoflurane produced a dose-dependent impairment of the auto-regulatory response to the angiotensin test as well as to the caval block. After hypotension, the autoregulatory response to increased blood pressure was restored within 15-25 min and to decreased blood pressures within 25-50 min. PMID- 15815331 TI - Changes in cerebral blood flow over time during isoflurane anesthesia in dogs. AB - The present study assessed the impact of time (6 h) on cerebral blood flow (CBF) during isoflurane anesthesia with and without vasopressor administration. All animals were prepared for measurement of CBF by the radiolabeled microsphere method under 1.4% end-tidal (1.0 MAC) isoflurane anesthesia. Surgery required 45 min and was followed by a 15 min stabilization period. In group 1 (n = 6), isoflurane 1.4% was administered for 6 h. CBF after 1 h of isoflurane was 92 +/- 9 ml/min/100 g (mean +/- SEM) and declined to 61 +/- 5 ml/min/100 g at 2 h and further declined to 37 +/- 5 ml/min/100 g at 6 hr. In group 2 (n = 6), isoflurane 1.4% was administered during the first hour. Thereafter, isoflurane 1.4% was continued, angiotensin II (0.3 mug/kg/min) was administered intravenously, and blood was withdrawn to maintain CPP constant for an additional 5 h, with hourly CBF determination. In this group, control CBF was 95 +/- 16 ml/min/100 g and flow was maintained at the control level through 4 h and then declined to 50 +/- 5 ml/min/100 g at 5 and 6 h. In group 3 (n = 6), 1.4% isoflurane was administered and phenylephrine (2.0 mug/kg/min) infusion was combined with hemorrhage to maintain control CPP in an identical sequence to group 2. In group 3, control CBF was 88 +/- 14 ml/min/100 g. As in group 1, CBF decreased significantly at 2 h (p < 0.05) to 68 +/- 13 ml/min/100 g and further declined to 49 +/- 7 ml/min/100 g at 6 h. In all three groups, CMRO2 remained at control levels and there were no changes in arterial carbon dioxide or CPP for the duration of the study. These data demonstrate that the hyperemia caused by isoflurane resolves over time during stable 1 MAC isoflurane anesthesia. The unanticipated interaction of angiotensin II and isoflurane producing a sustained cerebral hyperemia suggests that previous studies that used angiotensin II to support MABP during isoflurane may have reported the effects of angiotensin II in addition to or rather than the effects of isoflurane. PMID- 15815334 TI - On neuroanesthesia, intracranial pressure, and a dead horse. PMID- 15815332 TI - Repair of a cerebral artery aneurysm in a patient with persistent truncus arteriosus. AB - We describe the anesthetic management of a 19-year-old woman with persistent truncus arteriosus who presented for cerebral artery aneurysm clipping. The anatomy and pathophysiology of persistent truncus arteriosus are reviewed to provide a basis for the choice of anesthetic agents and monitoring devices in this patient. The hazards of induced hypotension in the patient with truncus arteriosus are also discussed. PMID- 15815335 TI - Another round of trivial pursuit? PMID- 15815336 TI - Transcranial Doppler sonography as a supplement in the detection of cerebral circulatory arrest. AB - The effects of compromised cerebral hemodynamics on intracranial blood flow velocity patterns (BFV) were studied by noninvasive transcranial Doppler sonography (TCD). Pulsatility index (PI) as an estimation of peripheral cerebral vascular resistance was additionally analyzed. TCD patterns were determined in 19 mechanically ventilated brain dead patients (group A) and 8 resuscitated or severely head injured patients (group B) with intermittent elevated intracranial pressure (ICP). Group A and B TCD data were compared to control measurements obtained in 20 healthy volunteers (group C). Normal TCD recordings (group C) were characterized by anterograde Doppler wave-forms with a predominantly high diastolic flow pattern and PI values of <1. In brain dead patients (group A), flow velocity profiles were significantly decreased. The flow curve consisted of low systolic anterograde spikes, while early diastolic flow components of low amplitudes revealed retrograde phases with a late diastolic zero flow. In some cases, total diastolic circulatory arrest was obtained. PI in brain dead patients was increased by a factor of 10 and was infinite in situations of zero net flow. The low systolic spikes and retrograde diastolic flow may be explained by the effects of reduced intracranial compliance due to increased intracranial pressure (ICP). The detection of bidirectional signals, indicating anterograde and retrograde movements of the blood column or low systolic spikes without diastolic signals, seems to be specific for the condition of brain death. Insonation of the basilar artery should be performed in order to exclude preserved perfusion of infratentorial brain regions during supratentorial circulatory arrest and vice versa. In group B patients, episodes of increased ICP predominantly affected the diastolic flow velocity component. In these patients, resistive Doppler signals with normal or reduced systolic peaks and decreased diastolic flow velocities were expressed as increases in PI. Noninvasive transcranial Doppler sonography appears to confirm cerebral circulatory arrest. Additionally, TCD may be of value for the early detection of impaired cerebral hemodynamics due to changes in intracranial compliance. PMID- 15815337 TI - Intensive care in the neurosurgical unit--a review of 2,492 patients. AB - The experience with 2,492 patients treated during a 9-year period in a neurosurgical intensive care unit (NICU) integrated into the ordinary neurosurgical wards is reported. The basic admission criterion was the need for ventilator treatment. During the period studied, the stay in the NICU decreased from a mean of 17 to 10 days. With respect to diagnosis, the largest group was patients with brain injuries (36%), followed by subarachnoid hemorrhage (24%), other intracranial bleeding (17%), and tumors (15%). Treatment consisted mainly of controlled ventilation, the duration of which ranged between 1 and 93 days with a mean of 5 days. Invasive monitoring was done relatively seldom, primarily due to a lack of equipment and staff. The incidence of nonneurosurgical complications was low, and the clinical outcome satisfactory. Nevertheless, it is concluded that for the best care of the severely ill neurosurgical patient, a special, well-equipped NICU with an adequate number of highly qualified staff members closely attached to the neurosurgical clinic is crucial. PMID- 15815338 TI - The effect of nitrous oxide on transcranial magnetic-induced electromyographic responses in the monkey. AB - Transcranial magnetic stimulation (TMS) to produce motor evoked potentials (MEPs) is a newly developed intraoperative modality to monitor functional integrity of the motor pathways. The present study was designed to examine the reliability of magnetic MEP recording under nitrous oxide (N2O) inhalation. Following ketamine injection (10 mg/kg i.m.) and endotracheal intubation, 16 monkeys were exposed to N2O:O2 mixture ratios 1:3, 1:1, and 3:1. Electromyographic (EMG) responses, evoked by pulsed magnetic fields applied extracranially to the scalp zone overlying motor cortex, were recorded from the contralateral fore- and hind limb flexor muscles. The scalp topography zone for contralateral muscle excitation was markedly reduced by 75 vol% N2O. Significant stimulation threshold elevation, latency prolongation, and amplitude depression were noted after inhalation of 75 vol% N2O (p <0.05) compared with 0.05) at each respective level of CO2 over the 3 h of the study. Cerebral metabolic rate for oxygen (CMRO2) was unchanged with time or CO2 alteration. CO2 responsivity (change in CBF/change in PaCO2) during hypercapnia was 0.8 +/- 0.2 ml min 100g mm Hg and during hypercapnia was 3.4 +/- 0.6 ml min 100g mm Hg at 1 h and both were unchanged from those value at 3 h. In animals receiving isoflurane and subjected to hypocapnia, total CBF during normocapnia at 1 h was 97 +/- 10 ml min 100 g and declined to 56 +/- 9 ml min 100 g at 3 hr (p <0.05). Over the same time period (1-3 h), hypocapnic CBF decreased from 44 + 5 to 27 +/- 3 ml min 100g (p <0.05). In animals receiving isoflurane and subjected to hypercapnia, normocapnic CBF decreased from 68 +/- 10 to 46 +/- 6 ml min 100 g at 3 h (p <0.05) and hypercapnic flow over the same time declined from 184 +/- 24 ml min 100 g to 135 +/- 18 ml min 100g (p <0.05). CMRO2 was not changed by either time or CO2 alteration. Between 1 and 3 h, CO2 responsivity during hypecapnia decreased from 4.1 +/- 0.9 to 1.8 +/- 0.4 ml min 100 g mm Hg (p <0.05). CO2 responsivity during hypocapnia decreased from over the same period decreased from 9.0 +/- 1.0 to 5.1 +/- 0.9 ml min 100 g mm Hg (p <0.05). Similar time-dependent trends were observed in most brain regions. We conclude that normocapnic CBF and cerebral CO2 responsivity decrease over time during isoflurane anesthesia and that these changes are not caused by changes in brain metabolism. PMID- 15815379 TI - The effect of an anesthetic induction dose of midazolam on motor potentials evoked by transcranial magnetic stimulation in the monkey. AB - The effect of a hypnotic dose (0.5 mg/kg) of midazolam (MDZ) on motor evoked potentials (MEPs) was examined in 12 monkeys. MEPs were elicited by transcranial magnetic stimulation (TMS) and the resultant potentials recorded from abductor pollicis brevis (APB) and anterior tibialis (AT) muscles contralateral to the stimulation site. After administration of MDZ, sequential MEP recordings were obtained at postinduction, hypnosis, awakening, emergence, and recovery periods. The results were compared with control values using one-way analysis of variance and Tukey's post-hoc test. Under hypnosis, MEP reproducibility was problematic as the potentials were occasionally ill identified and questionable. MDZ resulted in marked MEP scalp field reduction, coil demography alteration, stimulation threshold elevation, and amplitude suppression (p <0.01). Latency response was unaltered. During hypnosis, awakening, and recovery periods, the mean APB and AT thresholds were elevated by 39, 23, and 0% and by 60, 34, and 4% respectively; while APB and AT amplitudes were depressed by 95, 86, and 53% and by 99, 91, and 60%, respectively. We conclude that an induction dose of MDZ can produce profound and prolonged attenuation of TMS MEPs. The drug inhibitory effect on MEPs may persist after recovery. Anesthetic doses of MDZ should cautiously be used in the settings of MEP monitoring. PMID- 15815380 TI - The contributions of Sir William Macewen, a pioneer neurosurgeon, to an early quality assurance survey in anesthesia. PMID- 15815381 TI - Minor effect of lidocaine on brain electrical recovery after 5 minutes of almost complete cerebral ischemia in the rabbit. AB - In the present experiments, we tested the effect of a continuous infusion of low dose lidocaine on the time course of neuroelectrical recovery after an episode of almost complete ischemia in our single carotid artery model in the rabbit. In this preparation, carotid clamping elicited suppression of the electroencephalogram in less than 15 s, a drop in carotid stump pressure below 15 mm Hg, and evidence of minimal cerebral collateral perfusion in postmortem studies. We monitored mean arterial pressure, cerebral blood flow, carotid stump pressure, electroencephalogram, and auditory evoked potentials during three periods: control (30 min), ischemia (5 min), and reperfusion (90 min). Low-dose lidocaine (0.05 mg/kg/min) was continuously infused throughout the three periods. We specifically measured two peaks of the auditory evoked potentials and the most important finding was that the early wave (P1) partially recovered: to 35-40% in the control and to 64% in the lidocaine group (p <0.05 after 90 min). In contrast, the recovery of the later wave (P2) was very incomplete in both groups, and reached only 25-30% of the control amplitude. Similarly, the electroencephalogram of all of the rabbits failed to return to the control pattern but more animals in the lidocaine group (p <0.05) showed fewer pathological waves at the end of reperfusion. The present results indicate that a low dose of lidocaine had a small but significant action on electrical recovery after brief, almost complete global ischemia, confirming our previous observations on a rabbit model where the ischemic episode was less severe and the recovery more complete. These findings suggest that a low dose of lidocaine can accelerate the time course of neuroelectrical recovery after global ischemia and that the magnitude of this action depends very critically on the duration and severity of the ischemic event. PMID- 15815382 TI - Cerebrovascular effects of small volume resuscitation from hemorrhagic shock: comparison of hypertonic saline and concentrated hydroxyethyl starch in dogs. AB - To determine if hypertonic and hyperoncotic resuscitation solutions exerted comparable effects on cerebral hemodynamics following hemorrhagic shock, we compared randomly assigned, equal volumes (6.0 ml/kg) of hypertonic (7.2%) saline (HS) and hyperoncotic (20%) hydroxyethyl starch (HES) for resuscitation from acute experimental hemorrhage in 12 anesthetized dogs. Regional cerebral blood flow (radiolabeled microspheres), intracranial pressure (cisternal catheter), and systemic hemodynamics were recorded. Rapid hemorrhage reduced the mean arterial pressure to 45 mm Hg for 30 min. Resuscitation fluids were infused over 5 min. Both fluids restored mean arterial pressure and cardiac output equally. However, at 60 min following resuscitation, cardiac output decreased in the HS group in comparison to the HES group (1.7 +/- 0.1 vs. 3.1 +/- 0.2 L/min, p <0.05). Cardiac output rapidly declined, however, in the HS group in comparison to the HES group (p <0.05 60 min following resuscitation). Intracranial pressure and cerebral perfusion pressure were similar at all intervals. Regional cerebral blood flow was similar following both fluids. Neither fluid restored cerebral oxygen transport to baseline values. Based on these data, the authors conclude that, following severe hemorrhagic shock of brief duration, systemic and cerebral hemodynamic values are restored equally well by highly concentrated colloid or by hypertonic saline, although hypertonic saline only transiently improves cardiac output. PMID- 15815384 TI - Pulsatility index: an opinion. PMID- 15815385 TI - On high fashion and low power in anesthesiology research. PMID- 15815386 TI - Propofol vs. thiopental-isoflurane for neurosurgical anesthesia: comparison of hemodynamics, CSF pressure, and recovery. AB - Sixty otherwise healthy patients with no clinical signs of intracranial hypertension who were undergoing elective intracranial surgery were randomly assigned to receive anesthesia with either thiopental, 3-6 mg/kg i.v., and isoflurane, 0.5-1.5% (group 1, N = 30) or propofol, 1-2.5 mg/kg i.v., and propofol infusion, 40-200 microg/kg/h (group 2, N = 30). Both groups received 50% nitrous oxide in O2 subsequent to dural opening. During induction, the changes in heart rate (HR), mean arterial pressure (MAP), cerebrospinal fluid pressure (CSFP), and cerebral perfusion pressure (CPP) were similar between the groups, except at 3 min when the findings (mean +/- SEM) for CPP (81 +/- 3.3 vs. 70.3 +/- 2.8 mm Hg, p <0.05) were significantly lower in group 2. At intubation, the highest level of MAP (103.1 +/- 3.3 vs. 88.9 +/- 2.7 mm Hg, p <0.05) was significantly greater in group 1. At pinhead-holder application, the highest values of HR (81.8 +/- 3 vs. 73.9 +/- 2.1 beats/min, p <0.05), MAP (112.2 +/- 3.6 vs. 98.3 +/- 3 mm Hg, p <0.05), CSFP (15.2 +/- 1.3 vs. 11.6 +/- 1.1 mm Hg, p <0.05), and CPP (97.0 +/- 3.9 vs. 86.7 +/- 3.3 mm Hg, p <0.05) were significantly greater in group 1. During early (20-30 min) recovery, group 2 had higher Glasgow Coma Scale scores and a greater percentage of patients in whom eye opening, response to commands, extubation, speech, and time/space orientation were present. In conclusion, when compared to thiopentalisoflurane for intracranial surgery, propofol produces similar HR, MAP, CSFP, and CPP responses during induction, adequate control of these responses during nociceptive stimulation, and faster recovery for cerebral function postoperatively. PMID- 15815387 TI - The 21-aminosteroid U74006F does not markedly improve outcome from incomplete ischemia in the rat. AB - The 21-aminosteroid U74006F reportedly decreases neuronal injury following head injury or complete cerebral ischemia. We evaluated the ability of U74006F to improve outcome following incomplete cerebral ischemia in the rat. Ischemia was induced by right carotid occlusion combined with 30 min of hemorrhagic hypotension to 30 or 35 mm Hg. Animals in groups 1 and 2 were maintained on 1.4% isoflurane in room air and rats in groups 3 and 4 were ventilated with 70% nitrous oxide (N2O) and 30% oxygen. Rectal temperature was kept at 37C and PaCO2 and pH were maintained constant during ischemia. Group 1 (n = 10) and group 3 (n = 10) received a vehicle treatment. Group 2 (n = 10) and group 4 (n = 10) received 3 mg/kg of U74006F before ischemia and a 3-h infusion of 3 mg/kg/h of the drug after ischemia. Neurologic outcome was measured for 3 days and histopathology was evaluated at the end of the study. U74006F did not significantly improve neurologic outcome or histopathology during either isoflurane or N2O compared to the vehicle-treated groups. These results suggest that U74006F does not substantially inhibit ischemic damage produced in this model of incomplete cerebral ischemia. PMID- 15815388 TI - Intraarterial sodium amytal administration to guide preoperative embolization of cerebral arteriovenous malformations. AB - In preoperative embolization of cerebral arteriovenous malformation (AVM), one difficulty frequently encountered is assessment of the risk of ischemic deficit, posed by proceeding with embolization. One technique to assist prediction of tolerance of devascularization involves superselective injection of sodium amytal into cerebral arteries, prior to and during embolization. Since September, 1985, we have performed 119 embolization procedures in 77 patients with cerebral AVM. Of these, 89 procedures in 60 patients involved superselective intracerebral catheterization and embolization with either polyvinyl alcohol (PVA) alone or in combination with platinum microcoils, surgical silk, or a mixture of ethanol and microfibrillar collagen. In 50 of these procedures, we superselectively administered sodium amytal intraarterially immediately prior to embolization, for purposes of functional testing. Groups of patients undergoing testing were compared to groups not tested prior to embolization, for development of transient or permanent neurologic deficits or cerebral hemorrhage, subsequent to embolization. Significantly better results, both in absolute numbers and severity of complications, were found when testing was employed. No complications of amytal administration were seen. In four cases, embolization was altered or discontinued based on development of a deficit at the time of amytal administration. We conclude that the brief anesthetic effect of intraarterial testing with sodium amytal is a safe and important adjunct during preoperative embolization of cerebral AVMs. PMID- 15815389 TI - Intracranial volume-pressure relationship following flumazenil in anesthetized dogs. AB - A series of infusions of mock cerebrospinal fluid (CSF) was used to determine intracranial volume-pressure relationships in 12 anesthetized dogs. Measures of intracranial volume-pressure relationships included (a) CSF pressure prior to volume infusion (Po), (b) peak CSF pressure (Pp) caused by volume injection, (c) intracranial compliance [C, calculated as the ratio of change of intracranial volume (DeltaLV) to change of CSF pressure (DeltaLP)], (d) the volume pressure response (VPR, a measure of elastance, calculated as the ratio of DeltaLP to DeltaLV), (e) the pressure-volume index (PVI, calculated as the ratio of DeltaLV to log Pp/Po), and (f) estimated intracranial compliance (Ce, calculated from PVI as 0.4343PVI/Po). Measurements were made before giving flumazenil and after flumazenil doses of 0.0025 and 0.16 mg/kg in dogs receiving midazolam and in dogs not receiving midazolam (controls). The midazolam and control groups were examined both when CSF pressure was normal and when CSF pressure was increased to approximately 35 cm H2O. In dogs receiving midazolam at normal CSF pressure, 0.16 mg/kg of flumazenil (but not 0.0025 mg/kg) increased Po (by 4 +/- 2 cm H2O) and PVI, decreased Ce, and did not significantly changes C or VPR. Neither does of flumazenil caused consistent changes in dogs receiving midazolam when CSF pressure was increased prior to giving flumazenil, or in dogs not receiving midazolam. It is concluded that, in the presence of a benzodiazepine effect, large does of flumazenil increase CSF pressure from preflumazenil values and may be interpreted as worsening the intracranial volume-pressure relationship when the relationship is assessed by measures strongly affected by Po (PVI and Ce) but not when the relationship is assessed by indices that are relatively independent of Po (C and VPR). PMID- 15815390 TI - The effect of neuroleptanalgesia (droperiodol-fentanyl) on motor potentials evoked by transcranial magnetic stimulation in the monkey. AB - Motor evoked potentials (MEPs) after transcranial magnetic stimulation (TMS) have been examined in 12 monkeys under neuroleptanalgesia (NLA). Compound muscle action potentials were recorded from abductor policis brevis (APB) and gastroncnemius (GN) muscles contralateral to the stimulation site. After obtaining baseline tracings during emergence from methohexitone, 10 mg/kg i.m., NLA was induced using droperiodol, 0.3 mg/kg i.v. followed by fentanyl, 0.006 mg/kg i.v. Sequential MEP recordings were obtained 10 min after i.v. droperiodol, 2, 8, and 16 min after i.v. fentanyl, and during recovery. Replicable TMS MEPs were consistently recorded under NLA. However, droperidol and fentanyl caused significant stimulation threshold elevation, amplitude depression, and latency delay compared to control values (p <0.01). Ten minutes after droperiodol administration, the APB-GN threshold, amplitude, and latency values (mean +/- SD) were 0.81 +/- 0.2-0.84 +/- 0.1 T (baseline 0.57 +/- 0.1-0.59 +/- 0.1 T), 3.4 +/- 2.1-4.0 +/- 2.5 mV (baseline 8.0 +/- 3.7-9.0 +/- 3.7 mV), and 15.8 +/- 1.3-21.1 +/- 1.2 ms (baseline 14.9 +/- 1.2-20.1 +/- 1.3 ms), respectively. Addition of fentanyl resulted in further response deterioration. Two minutes after fentanyl injection, the APB-GN threshold, amplitude, and latency values were 0.88 +/- 0.18 0.95 %% 0.15 T, 2.1 +/- 1.7-2.0 +/- 2.1 mV, and 16.0 +/- 1.4-21.9 +/- 1.3 ms, respectively. Subsequent MEPs revealed gradual response improvement but, in contrast to baseline, remained markedly altered (p <0.05). During the recovery period (53 +/- 6 min), the APB-GN threshold, amplitude, and latency measurements were 0.66 +/- 0.1-0.77 +/- 0.2 T, 4.4 +/- 3.1-4.2 +/- 2.9 mV, 15.5 %% 1.4-20.9 +/ 1.7 ms, respectively. We conclude that, in a primate model, NLA maintains measurable TMS MEPs. Nevertheless, droperiodol and fentanyl produce significant and prolonged response alterations. Knowledge of these changes, while administering NLA drugs intraoperatively, is essential to interpretation of MEP data. PMID- 15815391 TI - Lidocaine accelerates recovery of brain electrical activity after 3 minutes of complete ischemia in the rabbit. AB - We recently reported that the continuous infusion of a low dose of lidocaine accelerated the recovery of the electroencephalogram and somatosensory evoked potentials after 20 min of incomplete cerebral ischemia in a single carotid artery preparation in the rabbit. In contrast, the recovery of the electroencephalogram and the auditory evoked potentials was minor on a comparable animal preparation subjected to 5 min of almost complete global ischemia. In the present study, we tested the hypothesis that the facilitatory effect of lidocaine on neuroelectrical recovery is of importance only during a certain combination of duration and severity of an ischemic episode. Similar to the previous studies, the rabbits had one carotid and both vertebral arteries permanently occluded and the other carotid artery reversibly clamped during the ischemic test. In the halothane-anesthetized animals, we monitored mean arterial pressure, carotid stump pressure, cerebral blood flow, auditory evoked potentials, and the electroencephalogram, before and for 90 min after 3 min of complete ischemia. The amplitude of the P1 wave (latency of 10 ms) of the auditory evoked potentials recovered more rapidly than on the previous 5 min preparation and returned towards the control amplitude more completely (p <0.05) in the lidocaine group. More importantly, the amplitude of the P2 wave (latency of 25 ms) recovered only 11% in the control group and 55% (p <0.05) in the lidocaine-infused animals. Comparable effects were observed in the early recovery of electroencephalographic activity during reperfusion. The present findings support the hypothesis that lidocaine may facilitate the recovery of the electroencephalogram and the evoked potentials only when an ischemic episode is below a certain combined level of duration and severity. The accelerating effect of lidocaine on postischemic neuroelectrical recovery was observed using a low dose of the drug, and it was not associated with changes in hemodynamic or cerebrovascular parameters or of blood composition. The latter suggests that the reported action may be related to some specific property of the local anesthetic action of lidocaine, such as blockade of axonal Na channels. PMID- 15815392 TI - Effect of massive intraoperative thiopental loading on cardiovascular hemodynamics and myocardial performance. AB - Anesthetics that protect the brain may also depress the heart. We recently anesthetized a patient with a giant cerebral aneurysm of the basilar artery who had it clipped during circulatory arrest. Thiopental loading was carried out prior to and during cardiopulmonary bypass, and we monitored transesophageal echocardiographic function and the hemodynamic profile throughout. The patient demonstrated little change in cardiovascular parameters or myocardial performance, and we conclude that the massive dose of thiopental was well tolerated. PMID- 15815396 TI - Controlled hypotension during neurosurgery: pro. PMID- 15815393 TI - Unexpected myocardial complications after controlled hypotension. AB - Two patients presented with serious myocardial ischemia after controlled hypotension for intracranial aneurysm clipping. Controlled hypotension has been associated with cardiovascular complications in the past. On the basis of their age and medical history, however, these two patients were considered to be at relatively low risk for cardiovascular complications from controlled hypotension, and thus minimal monitoring (i.e., ECG lead II and radial arterial blood pressure) was employed for detection of myocardial ischemia. If controlled hypotension is to be employed during intracranial aneurysm surgery, monitoring for intraoperative myocardial ischemia should be a major consideration of anesthetic management even in the apparently healthy patient. PMID- 15815397 TI - Controlled hypotension for cerebral aneurysm surgery: are the risks worth the benefits? PMID- 15815398 TI - The effect of evans blue dye on CMRO2 calculations: do two wrongs make a right? PMID- 15815418 TI - Parturition and intracranial pressure. PMID- 15815419 TI - The effects of pipecuronium bromide on intracranial pressure and cerebral perfusion pressure. AB - Twenty patients with expansive pathologic intracranial lesions, who were anesthetized with thiopental, nitrous oxide in oxygen, and fentanyl and mechanically ventilated to ensure normocarbia, received pipecuronium bromide 70 microg/kg i.v. Intracranial pressure (ICP), heart rate, arterial pressure, central venous pressure (CVP), EKG, and end-tidal CO2 were simultaneously recorded for 5 min before and for 15 min after administration of the muscle relaxant. No statistically significant changes in ICP and cerebral perfusion pressure were observed after administration of pipecuronium bromide. Cardiovascular stability was maintained during the study period except for a small, although significant, decrease of the CVP from 5.7 +/- 2.5 (SEM) to 5.0 +/ 2.5 mm Hg. These results, together with the long-lasting muscular effect of pipecuronium bromide, suggest that this new neuromuscular blocking agent may be used for muscle relaxation during neurosurgical operations in patients who have normal intracranial pressure at the time of administration of the drug. PMID- 15815420 TI - Effect of an anion transport inhibitor, L-644,711, on brain injury and edema after temporary middle cerebral artery occlusion in rats. AB - The affect of L-644,711, an anion transport inhibitor, on ischemic brain injury and edema was investigated. Spontaneously hypertensive rats were given one of the following doses of intrathecal L-644,711 during 180 min of middle cerebral artery occlusion and 120 min of reperfusion: control, vehicle only; dose I, 100 microg/kg: dose II, 200 microg/kg; dose III, 250 mug/kg; or dose IV, 320 microg/kg. Immediately after the 5-h period of ischemia and reperfusion, the brains were analyzed for brain injury with 2,3,5-triphenyltetrazolium chloride, and for edema by microgravimetry (specific gravity). There were no between-group differences in specific gravity (brain water content). Brain injury (% of the hemisphere ipsilateral to middle cerebral artery occlusion) was less (p <0.05) in rats that received the 250 (35 +/- 5%, mean +/- SD) or 320 microg/kg (36 +/- 6%) doses of L-644,711 vs. the control group (47 +/- 5%). L-644,711 has been hypothesized to affect brain injury by improving the neuronal acid-base state, inhibiting astroglial swelling, decreasing neutrophil aggregation, or reducing glutamate release. The microgravimetric data do not support astroglial swelling as a primary mechanism of decreased brain injury. PMID- 15815421 TI - Thiopental effect on cerebral blood flow during carotid endarterectomy. AB - To investigate the effect of thiopental on cerebral blood flow (CBF) during carotid endarterectomy, five patients receiving isoflurane-N2O anesthesia were studied. During the period of temporary bypass shunting, a baseline CBF was measured using i.v. Xe washout, and global CBF was calculated from the mean of 10 detectors. Thiopental was given in a dose sufficient (mean 4.5, range 2.6-5.8 mg/kg) to result in burst-suppression on the electroencephalogram (EEG) of approximately 1:1 duration and CBF was measured again. Data were compared using repeated measures analysis of variance. Thiopental significantly reduced mean (+/ SE) CBF (ml/100 g/min) from 37 +/- 6 to 18 +/- 2 (p <0.02). Corresponding PaCO2 (mm Hg) values were 42.8 +/- 1.2 and 41.2 +/- 1.6 and mean systemic blood pressure (mm Hg) was 101 +/- 3 and 100 +/- 6, respectively (NS). Mean % change in CBF was 48 +/- 5 (range 32-62%). There was no relationship between the dose administered and the change in CBF. During steady-state anesthesia, a small dose of thiopental capable of suppressing EEG resulted in a profound reduction in CBF. PMID- 15815422 TI - The effects of oxytocin on intracranial pressure. AB - The commonly used vasodilators sodium nitroprusside and nitroglycerine increase the intracranial pressure (ICP). Oxytocin is also a vasodilator although its primary effect is contraction of uterine smooth muscle. We therefore studied the effect of oxytocin on the ICP of cats. Twelve cats were assigned to two equal groups. One group had normal ICP and in the other the ICP was artificially increased by inflating a balloon placed in the extradural space. All of the cats initially received an oxytocin infusion for 15 min. After a period of stabilization, an additional dose of oxytocin was injected as a bolus. Oxytocin administered as an infusion did not change the ICP significantly in any of the cats. Oxytocin administered as a bolus increased the ICP from 27 +/- 4 to 31 +/- 2 mm Hg in the cats with artificially increased ICP. There were no significant changes in the mean blood pressure or heart rate in either group. PMID- 15815423 TI - Effects of ethanol on spinal cord blood flow in the rat. AB - This study examines the effects of low and high concentrations of ethanol on spinal cord blood flow (SCBF) in the rat. SCBF was measured in the following blood pressure ranges: (a) <60 mm Hg, (b) 60-90 mm Hg, (c) 90-120 mm Hg, (d) 120 150 mm Hg, and (e) >150 mm Hg. Rats were anesthetized with 1.4% isoflurane in air and randomly assigned to the following treatment groups: group 1 (n = 12), intraperitoneal (i.p.) saline injection; group 2 (n = 10), 1 g/kg of ethanol i.p.; and group 3 (n = 14), 4 g/kg of ethanol i.p. Blood pressure was increased by intravenous phenylephrine infusion or lowered by a combination of intravenous trimethaphan and blood withdrawal. The SCBF was measured in cervical, thoracic, and lumbar segments using radioactive microspheres. The plasma ethanol concentration was 0 mg/ml for group 1, 0.64 +/- 0.06 mg/ml (mean +/- SEM) in group 2, and 4.18 +/- 0.11 mg/ml in group 3. In control rats, the cervical SCBF was higher than the thoracic or lumbar SCBF, evaluated over the entire blood pressure range (analysis of variance, p <0.05). This difference in regional SCBF was abolished by ethanol. Ethanol produced a significant decrease in cervical and lumbar SCBF (p <0.05) but not thoracic SCBF (p = 0.07). This decrease in SCBF was most pronounced at high blood pressures. These results suggest that ethanol produces vasoconstriction in the spinal cord that is countered by autoregulatory vasodilation at low blood pressures. PMID- 15815424 TI - Detection of arterial gas embolism by increased end-tidal nitrogen in dogs. AB - Increased end-tidal (ET) nitrogen in a patient being ventilated with a nitrogen free gas mixture through a leak-free circuit has been considered a specific sign of venous air embolism. We hypothesized that increased ETN2 would occur after arterial air emboli, just as following venous air emboli, and that clinically relevant arterial air emboli could be detected with respiratory gas monitoring by mass spectrometry. After approval from the institutional Animal Utilization Committee, eight mongrel dogs were studied. All were anesthetized with pentobarbital and ventilated with room air by a volume ventilator. Each animal was monitored by a femoral artery and a pulmonary artery catheter for systemic and pulmonary blood pressures, respectively, an electrocardiograph, pulse oximetry, and inspired and expired respiratory gas measurements by mass spectrometry. Arterial blood gas analysis was undertaken after one series of air emboli. Air boluses (containing the nonradioactive nitrogen isotope N2) of 50, 100, 200, and 500 mul/kg were injected slowly into the distal aorta through a second arterial catheter advanced 35 cm above the inguinal ligament. All emboli >/=100 mul/kg and 60% of the 50 mul/kg emboli were detected by increased ETN2 within 30 s, reaching peak levels in <2.75 min. The washout time for the N2 was longer for larger emboli, ranging from 2.9 +/- 2.8 min for 50 mul/kg emboli to 17.3 +/- 3.2 min for the 500 mug/kg emboli. There were no significant changes in end-tidal carbon dioxide, pulmonary or systemic blood pressures, or arterial blood gases. Increased ETN2 can no longer be considered pathognomonic for venous air embolism; arterial air embolism may have occurred. PMID- 15815425 TI - Dose-related changes in the rate of CSF formation and resistance to reabsorption of CSF during administration of fentanyl, sufentanil, or alfentanil in dogs. AB - The rate of cerebrospinal fluid (CSF) formation (Vf) and resistance to reabsorption (Ra) of CSF were determined in dogs at four doses of fentanyl (0.05, 0.18, 0.60, and 3.0 microg.kg. min), sufentanil (0.01, 0.04, 0.13, and 0.60 microg.kg min) and aflentanil (1.4, 4.0, 13.0, and 40.0 microg.kg min). Results were compared within and between groups and to previously reported normal values (obtained during a variety of background anesthetics) for Vf (0.030-0.054 ml/min) and Ra (220-253 cm H2O ml min) in dogs. At the two lower doses of fentanyl and at all doses of sufentanil and alfentanil, Vf values were not significantly different from previously reported normal values. At the two higher doses of fentanyl, Vf decreased by 24 and 49%, respectively. At the two lower doses of all three drugs, Ra was significantly decreased, with mean values 40-52% below previously reported normal values. At the two higher doses of alfentanil, Ra values were not significantly different from previously reported normal values, and at the two higher doses of fentanyl and sufentanil, Ra was unchanged or increased. It is concluded that, among these three narcotics, reduction of CSF volume (as determined by the balance between Vf and Ra) is favored most by fentanyl, and ease of CSF volume contraction (as determined by Ra) is favored most by alfentanil. PMID- 15815426 TI - A comparison of the cerebral and hemodynamic effects of mannitol and hypertonic saline in a rabbit model of acute cryogenic brain injury. AB - There has recently been an increased interest in the use of hypertonic saline solutions in the fluid resuscitation of trauma victims and patients with uncontrollable intracranial hypertension. In this study, the cerebral and hemodynamic effects of 3.2% hypertonic saline solution were compared with those of an equiosmolar (20%) mannitol solution or 0.9% saline in a rabbit model of acute cryogenic brain injury. Forty-five minutes following the creation of a left hemispheric cryogenic brain lesion, equal volumes (10 ml/kg) of hypertonic saline, 0.9% saline, or mannitol were infused over a 5-min period. Monitored variables over the ensuing 120 min included mean arterial pressure, central venous pressure, intracranial pressure (ICP), hematocrit, and serum osmolality. At the conclusion of the 2-h study period, hemispheric water contents were determined by gravimetric analysis and the wet/dry weight method. There were no significant differences in mean arterial pressure between the three groups at any time during the experiment. Plasma osmolality was significantly increased by +/- 10 mOsm/kg following infusions in both the mannitol and hypertonic groups compared to the saline group. The infusion of either mannitol or hypertonic saline produced a transient and significant decrease in ICP during the first 60 90 min but not at 120 min after cryogenic brain lesion, whereas animals in the saline group demonstrated a continual increase in ICP. However, there appeared to be no significant differences in ICP between animals receiving mannitol or hypertonic saline at any time point following infusion of solutions. We conclude that following acute cryogenic brain injury, infusions of equal volumes of equiosmolar solutions of hypertonic saline or mannitol will transiently reduce ICP as compared to equal volumes of normal saline. However, hypertonic saline is not superior to mannitol in its ability to reduce ICP in this model of intracranial hypertension. PMID- 15815427 TI - Apneusis in a man with a supratentorial tumor. AB - A 67-year-old 163-cm tall man weighing 56 kg was scheduled for right fronto temporal craniotomy following a diagnosis of right supratentorial brain tumor. Prior to surgery he was somnolent, but response to verbal questioning was accurate. Heart rate was 50 beats/min, respiratory rate 20 breaths/ min, and blood pressure 148/80 mm Hg. Neurological examinations revealed a space-occupying lesion in the right frontal deep white matter and basal ganglia, and a moderate increase in intracranial pressure. There were no signs of brain-stem dysfunction. Upon arrival in the operating room and before the induction of anesthesia, the impedance pneumograph showed an apneustic respiratory pattern, which, along with bradycardia, persisted until the 5th postoperative day. A diagnosis of anaplastic astrocytoma of Grade III was made postoperatively. Usually we record the respiratory pattern in patients with brain injury or with brain tumor prior to surgery. We noted the apneustic pattern in one of 49 patients with head injuries and brainstem dysfunction and in one of 98 with brain tumor, but none of 55 medical student volunteers. The rate of occurrence of apneusis in patients with brain disease is 2 of 147 (1.36%). PMID- 15815430 TI - Hypertonic saline in head trauma: a new recipe for drying and salting. PMID- 15815428 TI - Cerebral oxygen metabolism measured during hypothermic circulatory arrest: a case report. AB - Infrared transmission spectroscopy was used to detect changes in cerebral oxy- and deoxyhemoglobin attenuation of infrared light in a patient undergoing circulatory arrest at 15 degrees C. Wavelength specific attenuation was used to determine hemoglobin oxygen saturation in the cerebral vasculature using a previously described paradigm. The fate of the residual oxygenated hemoglobin in the cerebrovascular compartment was followed during 15 min of circulatory arrest at 15 degrees C, and cerebral oxygen metabolism was identified as the progressive desaturation of this hemoglobin. The aggregate arterial and venous saturation fell from a normal value of 70% to an abnormally low value of 50%. Reperfusion resulted in near normalization of cerebrovascular oxygen saturation over 5 min, but evidence of persistent mild desaturation below prearrest baseline followed reperfusion. This case demonstrates both continued cerebral oxygen metabolism under hypothermic, circulatory arrest conditions and a cerebral oxygen delivery consumption mismatch during reperfusion. PMID- 15815431 TI - Hypertonic saline lowers raised intracranial pressure in children after head trauma. AB - Eighteen pediatric patients who sustained traumatic brain injury were enrolled in a double-blind, crossover study comparing the effects of 3% saline and 0.9% saline infusions on raised intracranial pressure (ICP). After resuscitation, each patient received a bolus of each saline concentration, and ICP was monitored for 2 h. Initial mean ICP before 0.9% saline infusions equaled 19.3 mm Hg and averaged 20.0 mm Hg during the subsequent 2-h trials (p = 0.32). Baseline mean ICP before 3% saline administration equaled 19.9 mm Hg and averaged 15.8 mm Hg for 2 h postinfusion (p = 0.003). Central venous pressure did not change significantly in either group, nor did measurements of renal function. Serum sodium concentrations increased in all 18 trials of 3% saline. Maximal concentrations of serum sodium occurred 30 min after bolus administration of 3% saline. Three percent saline significantly reduces raised ICP after traumatic brain injury when compared with normal saline. Intravascular dehydration, as measured by central venous pressure, did not occur during the study period. PMID- 15815432 TI - EEG-assisted titration of propofol infusion during neuroanesthesia: effect of nitrous oxide. AB - Total intravenous anesthesia (TIVA) with propofol is an alternative to standard techniques for neuroanesthesia. The present study compared the hemodynamic and recovery profiles of 46 neurosurgical patients randomly assigned to one of three different anesthetic treatment groups. Group 1 was anesthetized with a TIVA technique in which propofol was titrated using an EEG-assisted quantification method. Group 2 received a similar propofol-based infusion technique in combination with nitrous oxide. Group 3 (control) received a standard anesthetic technique consisting of thiopental, nitrous oxide, fentanyl, and isoflurane. Significantly less propofol was required in group 2 than in group 1 (7.4 +/- 1.9 vs. 9.0 +/- 1.0 mg/kg/h, respectively). The propofol blood concentration at the first appearance of EEG burst suppression was also higher in group 1 compared to group 2 (5.8 +/- 1.1 vs. 4.8 +/- 0.8 microg/ml). However, 25% of the patients in group 2 were treated for hypotension after induction, compared to none in groups 1 and 3. Hypertensive episodes, on the other hand, were more frequent in groups 1 (43%) and 3 (31%) than in group 2 (12%). Time to awakening was significantly shorter in the control group (6 +/- 6 min) than in groups 1 (14 +/- 10 min) or 2 (12 +/- 16 min). In conclusion, titration of propofol to achieve a burst suppressive EEG pattern resulted in a slower emergence from anesthesia than a standard "balanced" technique. Use of nitrous oxide with propofol produced more hypotension during induction; however, its use improved hemodynamic stability during the maintenance period. PMID- 15815433 TI - Elevated plasma levels of neuropeptide Y upon electrical stimulation of the sympathetic chain in humans. AB - Neuropeptide Y (NPY) a potent vasoconstrictor peptide, is co-stored with the classic neurotransmitter noradrenaline (NA) in certain peripheral sympathetic neurons and has been suggested to be a co-transmitter in vascular control. Surgical interruption of the sympathetic nerve supply of the upper extremity was performed to treat palmar hyperhidrosis in five patients. Intraoperative electrical stimulation (10 Hz; 0.5 ms; 0.5-3.0 mA) of the right and left sympathetic chains for 1 min was employed to determine the correct level for surgical excision. Blood pressure and heart rate were recorded continuously. Blood was sampled from the antecubital vein on the side subjected to stimulation for the determination of NPY-like immunoreactivity (LI) and plasma catecholamines. Blood samples were collected before and during stimulation, as well as 30 s and 2, 5, and 10 min after stimulation. A maximal blood pressure response was observed during the stimulation, but the magnitude varied markedly and was independent of the side on which the sympathetic chain was stimulated. During 3 of the 10 stimulations, the mean arterial blood pressure (MABP) increased 5-10 mm Hg, while in the other 7 an increase of 30-75 mm Hg was seen. No significant changes in heart rate or plasma adrenaline were found. The peak increase in NPY-LI plasma levels was noted 2 or 5 min after stimulation, while the corresponding peak for NA occurred during or 30 s after the stimulation. The maximal changes in plasma NPY-LI and NA were significantly correlated with the changes in blood pressure (NPY-LI, r = 0.80, p <0.01; NA, r = 0.84, p <0.01) as well as with each other (r = 0.95; p <0.001). It is concluded that, after electrical stimulation of the sympathetic chain in humans, plasma levels of both NPY-LI and NA rise, indicating release of these substances from sympathetic neurons. It is therefore possible that the vasoconstrictor peptide NPY is involved, together with NA, in the blood pressure response elicited by the stimulation. PMID- 15815434 TI - Plasma myeloperoxidase and vitamin E levels in head injury: preliminary results related to outcome. AB - This preliminary study was designed to assess a possible role of neutrophil activation and to determine the prognostic value of plasma myeloperoxidase (MPO) and vitamin E (Vit. E) levels in severe head injury. Plasma MPO and Vit. E levels were measured in nine severely head-injured patients (Glasgow Coma Score 160. Blood pressure was increased with phenylephrine or decreased with trimethaphan combined with blood withdrawal. Cerebral blood flow was not measured when MABP was less than 60 mm Hg in awake rats. Autoregulation was seen in all brain areas between 60 and 140 mm Hg in both treatment groups. Although regional cerebral blood flow was not different between the two treatment groups, PaCO2 was 2-4 mm Hg lower in awake rats. This suggests that PaCO2 corrected cerebral blood flow may be 10-20% lower with fentanyl-N2O anesthesia. PMID- 15815436 TI - The effects of propofol on cerebral blood flow in correlation to cerebral blood flow velocity in dogs. AB - This study correlates the effects of propofol on cerebral blood flow (CBF) and middle cerebral artery blood flow velocity in dogs. CBF was measured using radioactive microspheres. Cerebral oxygen consumption (CMRO2) was measured with each CBF determination. Blood flow velocity was measured through a transtemporal window using a pulsed 8 MHz transcranial Doppler ultrasound system (TCD). Electroencephalogram (EEG) was continuously recorded over both cerebral hemispheres. Cardiac output (CO) was measured using an electromagnetic flow probe placed on the pulmonary artery. Baseline measures were made in all dogs (n = 11) with 0.7% isoflurane end tidal and 50% N2O in O2. There were two treatment groups. In group 1 (n = 6), propofol (0.8 mg/kg/min) was infused and a second measurement made at induction of EEG burst suppression (12 +/- 2 min). CBF and CMRO2 decreased by 70% and mean blood flow velocity decreased by 60%. Blood pressure, heart rate, and CO did not change. Propofol infusion was discontinued and all parameters were measured following recovery of EEG to baseline activity (48 +/- 9 min). CBF and blood flow velocity increased 35 and 25%, respectively, and CMRO2 increased by 32% during this period. A second propofol infusion (0.8 mg/kg/min) was started and all cerebral and systemic hemodynamic parameters were again determined at induction of EEG burst suppression (12 +/- 2 min). CBF decreased 35% and blood flow velocity decreased 25% to levels seen during the first propofol infusion. Over the entire study, changes in CBF correlated with changes in blood flow velocity (r = 0.86, p < 0.05). In group 2 (n = 5), four control measures were made at the same time intervals as in group 1. Baseline CBF and blood flow velocity were lower in group 2 compared to group 1 but these measures did not change over time. Our results show that propofol produces marked decreases in CBF in dogs and that these changes are closely correlated with CBF velocity. PMID- 15815437 TI - Anesthetic implications for children undergoing computerized tomography-guided stereotactic brain biopsy. AB - Anesthesiologists spend much of their time administering anesthesia to children outside of the operating room, especially for neuroradiological evaluation. Neuroradiological procedures include magnetic resonance imaging (MRI), computed tomography (CT), positron emission tomography, myelography, pneumoencephalography, and arteriography. Each of these procedures presents a "working environment" that compromises the ability to deliver a safe and well monitored anesthetic (1). CT-guided stereotactic brain biopsy in children requires two anesthetic locations (2). The stereotactic guidance apparatus renders the airway inaccessible both during CT and during transport to the operating room. We report a case of a child undergoing CT-guided stereotactic brain biopsy to elucidate the anesthetic considerations of this logistically difficult procedure. PMID- 15815438 TI - Case report acute respiratory failure after posterior cranial fossa surgery: report of two cases with unusual etiology. AB - Acute respiratory failure (ARF) occurred in two patients who had normal preoperative findings and intraoperative courses, early after posterior cranial fossa (PCF) surgery. In both instances, the postoperative pulmonary failure appeared to be due to an unknown pulmonary cancer, revealed by autopsy. The diagnosis of an asymptomatic pulmonary malignancy might be considered in the presence of ARF of unclear etiology in patients who undergo PCF surgery for removal of a brain tumor. PMID- 15815441 TI - Routine barbiturate brain protection during cardiopulmonary bypass cannot be recommended. PMID- 15815442 TI - Cerebral protection during cardiac surgery: ban the barbiturates? PMID- 15815443 TI - American board of anesthesiology-the sequence of events. PMID- 15815444 TI - A protocol for intraoperative somatosensory (SEP) and motor evoked potentials (MEP) recordings. PMID- 15815445 TI - Propofol: an intravenous anesthetic for neuroanesthetic practice? PMID- 15815446 TI - Focal cerebral ischemia in rats: effect of phenylephrine-induced hypertension during reperfusion. AB - After 180 min of temporary middle cerebral artery occlusion in spontaneously hypertensive rats, the effect of phenylephrine-induced hypertension on ischemic brain injury and blood-brain barrier permeability was determined. Blood pressure was manipulated by one of the following schedules during 120 min of reperfusion: Control, normotensive reperfusion; 90/hypertension (90/HTN), blood pressure was increased by 35 mm Hg during the initial 90 min of reperfusion only; 15/hypertension (15/HTN), normotensive reperfusion for 30 min followed by 15 min of hypertension and 75 min of normotension. Part A, for eight rats in each group brain injury was evaluated by staining tissue using 2,3,5-triphenyltetrazolium chloride and edema was evaluated by microgravimetry. Part B, for eight different rats in each group blood-brain barrier permeability was evaluated by measuring the amount and extent of extravasation of Evans Blue dye. Brain injury (percentage of the ischemic hemisphere) was less in the 15/HTN group (16 +/- 6, mean +/- SD) versus the 90/HTN group (30 +/- 6), which was in turn less than the control group (42 +/- 5). Specific gravity was greater in the 15/HTN group (1.043 +/- 0.002) versus the 90/HTN (1.036 +/- 0.003) and control (1.037 +/- 0.003) groups. Evans Blue (mug g-1 of brain tissue) was greater in the 90/HTN group (24.4 +/- 6.0) versus the control group (12.3 +/- 4.1), which was in turn greater than the 15/HTN group (7.3 +/- 3.2). This study supports a hypothesis that during reperfusion, a short interval of hypertension decreases brain injury and edema; and that sustained hypertension increases the risk of vasogenic edema. PMID- 15815447 TI - Human brain temperature during anesthesia for intracranial operations. AB - The intraventricular and rectal temperatures were registered in nine patients subjected to major surgery of the brain. Copper-constantan thermocouples were introduced into the lumen of an intraventricular catheter also used for perioperative monitoring of intracranial pressure. During anesthesia, the intraventricular temperature was higher than rectal temperature, the mean difference being 0.30 +/- 0.24 degrees C. No significant changes in intraventricular temperature were seen during different stages of the operations. It is concluded that during routine anesthesia rectal temperature can be relied on for a reasonable estimation of human brain temperature. It should be observed, however, that in the postoperative period, both rectal and intraventricular temperature rose considerably. In three patients, the intraventricular temperature rose as much as 2.5-4 degrees C, thus increasing the temperature gradient between rectum and brain. The relevance of these findings are discussed. PMID- 15815448 TI - In vivo effect of calcitonin gene related peptide on middle cerebral artery blood flow velocity in humans. AB - A double blind, placebo controlled, escalating dose study was undertaken in five healthy, conscious volunteers to investigate the effects of human synthetic alpha calcitonin gene-related peptide (CGRP) on middle cerebral artery (MCA) blood flow velocity using transcranial Doppler sonography. During placebo infusion, there was no significant change in any of the parameters studied. During CGRP infusion, all subjects showed flushing of the face and neck. Infusion of CGRP caused a significant increase in arterial pulse pressure and heart rate and a fall in diastolic and mean arterial blood pressure when compared to baseline. Peak and mean MCA velocity did not change significantly. There was a significant increase in pulsatility index though interpretation of this was confounded by the central systemic effects of CGRP. The observed haemodynamic changes may be explained by a prompt sympathetic nervous system response in order to maintain mean arterial blood pressure. Our data suggest that if cerebral vasodilatation had occurred, it was not associated with increased blood flow as an increase in MCA velocity might have been expected. An alternative explanation is that, at the doses employed, CGRP did not cause dilatation of normal cerebral vessels in healthy subjects. This does not however exclude the fact that CGRP may cause dilatation of a spastic artery as seen in cerebral vasospasm following subarachnoid haemorrhage. PMID- 15815449 TI - Electroencephalogram, cerebral metabolic, and vascular responses to propofol anesthesia in dogs. AB - Previous studies on the cerebral effects of propofol report conflicting results regarding the cerebral metabolic rate for oxygen (CMRO2), cerebral blood flow (CBF), autoregulation of CBF, intracranial pressure, and cerebral perfusion pressure (CPP). The present studies were designed to examine these issues as well as propofol effects on the CBF responses to hypocapnia and on the electroencephalogram (EEG) in a well-known canine model that permits continuous determination of EEG activity, CMRO2, CBF, and cerebrospinal fluid (CSF) pressure. Dogs were studied at normocapnia (n = 6) and at hypocapnia (n = 6) during three doses of propofol (12, 24, and 48 mg kg(-1) h(-1)) and during a combination of propofol and elevated (20-25 mm Hg) CSF pressure. In both groups propofol caused dose-related decreases of EEG power and number of waveforms, CMRO2 (by 25-30%), and CBF (by 73-76%). The cerebral vasoconstrictor response to hypocapnia was preserved at all three doses of propofol. Autoregulation of CBF was preserved at the low and moderate doses of propofol but was impaired at the high dose of propofol (where CPP decreased significantly to approximately 41 +/- 13 mm Hg) and at the high dose of propofol combined with elevated CSF pressure (where CPP decreased significantly to approximately 32 +/- 12 mm Hg). Cerebrospinal fluid pressure decreased (by 33-42%) when the continuous infusion of propofol was begun, but returned to prepropofol values as infusion of propofol continued. The authors conclude that low and moderate doses of propofol decrease EEG activity and CMRO2, causing an associated decrease of CBF and CSF pressure. Autoregulation of CBF and cerebral vascular CO2 reactivity are preserved at these propofol doses. In contrast, high dose propofol significantly decreases CPP, resulting in impaired autoregulation of CBF. PMID- 15815450 TI - A dose-response study of the influence of propofol on cerebral blood flow, metabolism and the electroencephalogram in the rabbit. AB - This experiment was designed to study the effect of progressively increasing blood propofol concentrations on cerebral blood flow (CBF), cerebral metabolic rate for oxygen (CMRO2), and the electroencephalogram (EEG). Nine New Zealand White rabbits were anesthetized with morphine (10 mg kg bolus and 2 mg/kg/h infusion) and 70% N2O. Both normothermia and normocarbia were maintained throughout the study. A 300 mum diameter platinum electrode and a 20 gauge sampling needle were inserted into the confluence of venous sinuses to permit the measurement of forebrain CBF and CMRO2. Cerebral blood flow was determined using the H2 clearance method and CMRO2 was calculated as CBF x arteriovenous O2 content difference. A single bifrontal EEG signal was also recorded. After baseline data were collected, a propofol infusion was begun, and the dose increased in a stepwise fashion from 0.28 to 1.11 mg/kg/min over 90 min (total dose 62.6 mg/kg). Every 22.5 min CBF, CMRO2, and EEG were recorded and arterial blood was sampled for the determination of propofol concentrations (by high performance liquid chromatography). Angiotensin II was used to maintain mean arterial pressure >/=80 mm Hg. Eight animals completed the protocol. Blood propofol concentrations rose progressively in all animals, reaching a mean of 34 +/- 12 microg/ml (+/-SD) at the end of 90 min. Electroencephalogram changes during the early stages of the infusion were extremely variable. However, concentrations above approximately 20 microg/ml were associated with progressive EEG suppression, and isoelectricity developed in two animals, at blood concentrations of 41 and 52 microg/ml. There was a progressive dose-related decrease in CBF, which reached a value of approximately 62% of baseline at a concentration of 40 microg/ml (as predicted by a polynomial curve fitted to a plot of CBF versus blood concentration). The CMRO2 also decreased progressively, reaching approximately 57% of baseline at 40 microg/ml. A plot of CMRO2 versus EEG total power suggests that isoelectricity should be associated with a CMRO2 approximately equal to 53% of baseline. We conclude that propofol produces a dose related decrease in CBF and CMRO2. The relationship between EEG suppression and CMRO2 is qualitatively similar to that seen with barbiturates. PMID- 15815451 TI - Autoregulation of cerebral blood flow in response to adenosine-induced hypotension in dogs. AB - During induced hypotension for surgical procedures, cerebral blood flow (CBF) autoregulation and cerebrovascular responsivity to CO2 may be impaired-changes that appear to be agent-specific. Adenosine is a potent endogenous systemic vasodilator and has been investigated as a hypotensive agent. In this study in dogs we investigated cerebral vascular responses to graded decreases of cerebral perfusion pressure (CPP) (100%, 60%, 45%, and 35% of control CPP) during normocapnia (PaCO2 = 37 mm Hg) and hypocapnia (PaCO2 = 21 mm Hg). CBF was measured using the venous outflow technique. Six mongrel dogs were anesthetized with halothane (0.6% inspired) and nitrous oxide (70%) in oxygen and studied during both normocapnic and hypocapnic hypotension. The entry sequence was randomized with >/= 1 h of recovery between normocapnia and hypocapnia. Hypocapnia reduced control CBF from 60.6 +/- 7.1 to 45.1 +/- 5.4 ml 100 g min (mean +/- SEM, p <0.05) during normotension. CBF was unchanged from control values during both graded normocapnic and hypocapnic hypotension until CPP reached 60% of control CPP (50 and 47 mm Hg for normocapnia and hypocapnia, respectively). Thereafter CBF decreased significantly from control values at 45% (37 mm Hg for both groups) and 35% (29 mm Hg for both groups) of control CPP. The lower limit of CBF autoregulation derived by applying linear regression analysis to the CBF-CPP relationship above and below the inflexion point was similar under both experimental conditions (60 +/- 1% of control CPP during normocapnia and 63 +/- 3% of control CPP during hypocapnia). CBF was significantly greater during normocapnia compared with hypocapnia at all levels of CPP, except at 35% of control when the values were similar. Cerebral metabolic rate was unchanged throughout the study. We conclude that neither CBF nor CO2 responsivity is appreciably altered during adenosine-induced hypotension when GPP remains above the lower limit of autoregulation of CBF. PMID- 15815452 TI - Disseminated intravascular coagulation in neurosurgical patients: diagnosis by new laboratory methods. AB - Disseminated intravascular coagulation (DIC) constitutes a part of the multiple organ failure (MOF) syndrome seen with such disorders as trauma and sepsis. Early detection of increased coagulation and fibrinolytic activity is important. The dynamic changes in some markers for early detection of the activation of these cascade systems are presented in relation to two patients with brain trauma. The clinical status and the severity of the disease were assessed by an established scoring method (APACHE II). The coagulation activation was noted by the appearance of increased end products of the coagulation cascade, such as soluble fibrin, thrombin-antithrombin complex, and prothrombin fragment 1 + 2. Fibrinolytic activation and an increased secondary inhibition of fibrinolysis were detected by increased levels of D-dimer and plasminogen activator inhibitor 1. Leukocyte activation was indicated by a rise in elastase. The laboratory results normalized with clinical improvement. These new methods seem to detect DIC earlier than traditional methods and may also be of value for monitoring treatment. PMID- 15815453 TI - An uncommon electrocardiographic change in subarachnoid hemorrhage: increase in the Sokolow's Index. AB - In this paper we are reporting a case of electrical left ventricular hypertrophy with increased Sokolow's index following subarachnoid hemorrhage. Two-dimensional echocardiography ruled out anatomical left ventricular hypertrophy, and the Sokolow's index eventually reverted to normal. This electrocardiographic abnormality has rarely been associated with subarachnoid hemorrhage and does not appear to be related to the neurologic grade of the patient because he presented with headaches only and was conscious throughout (Hunt and Hess grade I). The possibility that such electrocardiographic changes should reflect anatomical changes in the heart should always be ruled out by proper investigations (i.e., echocardiography and coronarography), as the presence of cardiac disorders would greatly influence medical and surgical management. In terms of electrocardiography, this observation suggests that the Sokolow's index is a poor indicator of left ventricular hypertrophy, and tends to reflect a more comprehensive catecholaminergic process. PMID- 15815454 TI - Delayed postoperative nonembolic neurologic deficit after left internal carotid endarterectomy in presence of known contralateral carotid artery occlusion. AB - Neurologic sequelae are known complications of carotid endarterectomy. The current overall perioperative stroke rate is 2-5% (1,2). The incidence of perioperative morbidity and mortality is increased in certain patient subgroups, including neurologically unstable patients and patients who have known contralateral common or internal carotid artery occlusion (3-9). We present the case of a patient who had known total right carotid artery occlusion and a known previous right cerebrovascular accident who sustained a contralateral deficit after a left internal carotid thromboendarterectomy with vein patch angioplasty. PMID- 15815458 TI - Inhaled lidocaine for prevention of hemodynamic changes in laryngoscopy and intubation. AB - This study was undertaken to examine the efficacy of lidocaine aerosol pretreatment in attenuating hemodynamic (HD) responses secondary to laryngoscopy and tracheal intubation in patients undergoing surgery for intracranial space occupying lesions (ICSOL). A semiclosed breathing system was improvised to generate aerosol of consistent density. Five percent lidocaine was nebulized in two different dosages (0.2 and 0.1 ml/kg in groups A and B, respectively); group C (control) received aerosol of normal saline. The average aerosol-treatment time was 24, 12, and 16 min in groups A, B, and C, respectively. Changes in heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), and rate pressure product (RPP) were analyzed by analysis of variance (ANOVA). In group A, tracheal intubation did not cause significant HD changes. In group B, a significant increase was observed in each HD parameter which, when compared with control, was less severe. Lidocaine toxicity, regurgitation, nausea, vomiting, or aspiration did not occur in any patient. Patients accepted the procedure well. This study found efficacy of the technique to be related to duration of aerosol treatment. PMID- 15815459 TI - Interatrial pressure difference during inflation of pneumatic antishock garments and positive end-expiratory pressure ventilation. AB - Since an increase in the right atrial pressure (RAP) above the left atrial pressure (LAP) may then lead to paradoxical air embolism, the efficacy of pneumatic antishock garments (PASGs) and ventilation with positive end-expiratory pressure (PEEP) in preventing increases in the RAP above the LAP was investigated during neurosurgical procedures. We examined the RAP and pulmonary capillary wedge pressure (PCWP) of 25 patients during neurosurgery of the posterior fossa. Each set of measurements of PCWP and RAP included measurements during induction of anesthesia in the supine position and the seated position prior to surgery. The PCWP and RAP were recorded during PEEP = 0 cm H2O, PEEP = 10 cm H2O, or with PASG inflation to pressures ranging from 40 to 80 mm Hg. The RAP increased from 8 +/- 6 to 20 +/- 7 mm Hg (PASG) and to 11 +/- 5 mm Hg (PEEP) in the seated position without a significant (p >0.05) change in the PCWP-RAP difference under any of the above conditions. We conclude that PASGs are a safer and more effective means of raising the RAP than PEEP ventilation during seated neurosurgical procedures without the danger of paradoxical air embolism. PMID- 15815460 TI - Cerebral blood flow measurements during blood pressure control with intravenous labetalol following craniotomy. AB - Cerebral blood flow measurements using a thermal diffusion technique were made in conjunction with an extensive cardiovascular evaluation, during and after administration of intravenous labetalol given for blood pressure control in craniotomy patients. Eighteen patients, ages 30-65 years, ASAII and III, scheduled for elective craniotomy, became hypertensive during emergence and recovery from a pentothal/fentanyl/vecuronium/N2O/isoflurane general anesthesia. Labetalol was administered in a stepwise manner every 10 minutes during an average period of 1 h. After this titration period, an 8-h maintenance period followed. During titration and maintenance periods, comprehensive systemic hemodynamic parameters were collected through intra-arterial and flow-directed pulmonary artery catheters. Cerebral cortical blood flow (CBF) was continuously recorded using a thermal diffusion cortical blood flow probe (Saber System). Data were analyzed using variance F tests to evaluate changes from baseline over time. Labetalol controlled postoperative hypertension in all cases with a total dose range of 0.4-6.8 mg/kg. During titration, statistically significant decreases in blood pressure were obtained, accompanied by a small decrease in systemic vascular resistance (SVR) and slight increase in cardiac index (CI). Heart rate decreased in a manner directly proportional to the dose of labetalol administered. In the maintenance period, further decreases in blood pressure and heart rate were observed, with significant decreases in central venous pressure, pulmonary capillary wedge pressure, and SVR and an increase in CI. All values remained within normal ranges and no adverse effects were observed. CBF decreased slightly during the study period, although not significantly (from 67 +/- 8 to 57 +/- 7 ml 100 g min). Blood pressure control achieved with labetalol in postoperative neurosurgical patients seems to be the result of mild alpha adrenoceptor blocking effects (i.e., reduced SVR) and beta-adrenoceptor blocking effects (i.e., reduced heart rate) at higher doses. The 6-8 h duration of effect of labetalol was enough to control postoperative hypertension in all patients with no additional therapy. Compared with alternative drugs available for blood pressure control in similar clinical conditions, labetalol appears to be reliable, safe, and effective, by providing a lasting effect with no evidence of rebound hypertension, increased CBF, or cardiac dysfunction. PMID- 15815461 TI - Anesthesia for selective dorsal rhizotomy in children. AB - Anesthetic records for the first 71 children who underwent selective dorsal rhizotomy (SDR) at our hospital were reviewed. Anesthesia during SDR must preserve muscle contraction in response to direct electrical stimulation of the dorsal nerve rootlets. In our experience, halothane, isoflurane, and narcotics do not interfere with electrophysiologic monitoring, even though relatively large doses are required during SDR. Propofol proved to be unacceptable as an anesthetic because of severe muscle spasms during electrical stimulation of the nerve rootlets. The body temperature rises predictably during the stimulation phase of SDR and active warming measures should be avoided. PMID- 15815462 TI - Cerebral blood flow at constant cerebral perfusion pressure but changing arterial and intracranial pressure: relationship to autoregulation. AB - Therapeutic agents for reducing raised intracranial pressure (ICP) may do so at the expense of reduced mean arterial pressure (MAP). As a consequence, cerebral perfusion pressure (CPP) = (MAP - ICP) may not improve. It is unknown whether the level of MAP alters cerebral blood flow (CBF) when MAP and ICP change in parallel so that CPP remains constant. This study investigates CBF at a constant CPP but varying levels of MAP and ICP in 12 anaesthetized cats. CBF was studied at three levels of CPP: 60 (n = 4), 50 (n = 4), and 40 mm Hg (n = 4) under conditions of both intact and impaired autoregulation. At CPP levels of 50 and 60 mm Hg, when autoregulation was intact, CBF remained unchanged. With loss of autoregulation, there was a trend for CBF to increase as MAP and ICP were increased in parallel at a CPP of 50 and 60 mm Hg, although the relationship did not achieve statistical significance. Absolute CBF levels were, however, significantly different between the autoregulating and nonautoregulating groups (p <0.001). At a CPP of 40 mm Hg, CBF showed a linear correlation with blood pressure (BP) (r = 0.57, p <0.05). These results demonstrate that when autoregulation is impaired, there is a functional difference between autoregulating and nonautoregulating cerebral vessels despite similar MAP and CPP. These results also show that at a CPP of 40 mm Hg when autoregulation is impaired, CBF depends more on arterial driving pressure than on CPP. PMID- 15815463 TI - Effect of sevoflurane on intracranial pressure, sagittal sinus pressure, and the intracranial volume-pressure relation in cats. AB - The effect of sevoflurane on intracranial pressure (ICP), sagittal sinus pressure (SSP), and the intracranial volume-pressure (V-P) relation was examined in cats. In experiment 1, on nine cats, changes in ICP and SSP were studied for 180 min during anesthesia with 1 MAC sevoflurane (2.6%, inspired) and 50% nitrous oxide (N2O) in oxygen (O2). ICP significantly (p <0.01) increased from 8.4 +/- 3.8 cm H2O (mean +/- SD), the control level to 10.6 +/- 5.1 cm H2O immediately after the administration of sevoflurane. ICP was unchanged for the subsequent 120 min but then increased significantly (p <0.05) 140 min after administration, being 15.5 +/- 9.0 cm H2O at 180 min. There were no changes in SSP or blood gases. In experiment 2, the rapid injection technique of mock cerebrospinal fluid was used to determine the intracranial V-P relation in ten cats. Measures of V-P relationships included (a) ICP before volume injection (Po), (b) peak ICP caused by volume injection (Pp), (c) intracranial compliance (C) calculated as the ratio of change of intracranial volume Delta V) to change of ICP (Delta P), and (d) the pressure volume index (PVI) calculated as the ratio of Delta V to log Pp/Po. The subjects were divided into two groups, one administered 2.6% sevoflurane and 50% N2O in O2 (n = 6) and the other 50% N2O in O2 (n = 4). Each cat in both groups was given two bolus injections into the lateral ventricle at 180 min after the start of anesthesia. Then, C and PVI were calculated. C and PVI in the group treated with sevoflurane were significantly (p <0.05) lower than in the other group. These findings suggest that prolonged use of sevoflurane increases the intracranial elastance. PMID- 15815464 TI - Airway obstruction after anterior cervical spine surgery. AB - Complications arising from operations on the cervical spine through an anterior approach are relatively uncommon. However, the surgical territory is such that when complications do arise they can be serious and occasionally life threatening. We report the case of a patient who developed acute respiratory obstruction after Cloward's procedure. PMID- 15815467 TI - Hypothermia but not NMDA receptor antagonist MK-801 attenuates neuron damage: are the results conclusive? PMID- 15815468 TI - Rekindle an old technique: spinal anesthesia for lumbar laminectomies. PMID- 15815471 TI - Ketamine decreases cerebral infarct volume and improves neurological outcome following experimental head trauma in rats. AB - In brain injury, concentrations of extracellular excitatory amino acids are increased and stimulate glutamate receptors in general and the N-methyl-D aspartate (NMDA)-preferring subtype in particular. That stimulation causes substantial calcium influx, which appears to initiate a cascade of events leading to neuronal death. Blockage of NMDA receptors with specific antagonists or noncompetitive ion channel blockers provides protection against excitatory amino acid-induced neurotoxicity. We previously reported that the NMDA receptor antagonist dizocilpine maleate improved the neurological severity score (NSS) after head trauma in rats. The present study was designed to determine whether ketamine, a NMDA receptor antagonist like dizocilpine maleate, improves neurological outcome following head trauma in rats. Thirty-two male Sprague Dawley rats (235-250 g) were divided into four groups. Groups A and B were surgically prepared only. Groups C and D were surgically prepared and then a nonpenetrating impact was delivered to the cranium over the left hemisphere. Groups A and C received no treatment. Groups B and D were treated with ketamine, 180 mg/kg i.p., 1 h after head trauma. The NSS was determined at 1, 2, 4, 10, 24, and 48 h following head trauma. After killing at 48 h, cortical slices were taken adjacent to the lesion on the traumatized hemisphere and from comparable sites on the nontraumatized hemisphere to measure the tissue specific gravity and water content. Brains were then placed in 4% formaldehyde and the volume of hemorrhagic necrosis measured 4 days later. Head trauma increased the NSS and, in the traumatized hemisphere, decreased the specific gravity, increased the water content, and caused cerebral infarction. With ketamine, the NSS at 24 and 48 h following head trauma was 7.4 +/- 2.6 and 6.7 +/- 2.6 (mean +/- SEM), respectively, significantly improved compared to the NSS in the untreated group of 12.6 +/- 2.6 and 11.3 +/- 2.6, respectively (p <0.02, Mann-Whitney U test). With ketamine, the volume of hemorrhagic necrosis was 88.0 +/- 23.1 mm, significantly less than that in the untreated group (147.4 +/- 22.4 mm; p <0.05, unpaired t test). The brain tissue specific gravity and water content at 48 h and the rectal temperature at 4 and 48 h after head trauma were not significantly different between treated and untreated groups. It is concluded that in this model of closed cranial impact, ketamine improves neurological outcome and decreases the volume of hemorrhagic necrosis without altering brain edema. PMID- 15815472 TI - End-tidal carbon dioxide as an indicator of arterial carbon dioxide in neurointensive care patients. AB - The relationship between the arterial partial pressure of carbon dioxide (Paco2) and the end-tidal carbon dioxide partial pressure (PEtco2) was evaluated in 11 critically ill adult neurointensive care patients during mechanical ventilation. It was hypothesized that the Paco2 to PEtco2 gradient, or P(a-Et)co2, was maintained and that PEtco2 can be used to determine Paco2 accurately in these patients. After approval by the Clinical Investigations Committee, when clinically indicated arterial blood gases (with Paco2) were measured, the PEtco2 was determined from the capnograph (Hewlett Packard 78520A infrared capnometer). The P(a-Et)co2 was evaluated for possible effects from changes in the other monitored hemodynamic and respiratory parameters. Linear regression analysis was used to determine the significance of the relationship between Paco2 and PEtco2 and other assessed variables. Student's t tests were used where applicable. A p value 0.05), the consistency and magnitude of the increase along with the likelihood of a type II error led us to conclude that (a) both DO and DA are less effective in monkeys than in humans and (b) whereas DO at all doses tested showed no indication of affecting CBF and CMR, DA increased CBF and CMR by 20 to 30% at a dose of 100 microg/kg/min. PMID- 15815474 TI - Effect of stable xenon inhalation on internal carotid artery blood flow in unanesthetized monkeys. AB - Stable xenon (Xe) gas, at inspired concentrations above 30%, reportedly increased cerebral blood flow (CBF) in animals and humans. An unpredictable Xe-induced elevation of CBF could result in erroneous CBF values being measured by Xe enhanced computed tomography (Xe-CT). In order to detect a potentially rapid and transient effect of Xe on CBF, estimations of supratentorial CBF were obtained by Doppler flow probes chronically and bilaterally implanted on the internal carotid arteries of five adult monkeys. The unanesthetized monkeys with a clear plastic helmet were equilibrated for 15 min on a control gas (33% N2/67% O2) randomly exposed for 5 min to gas mixtures of either 33% Xe/67% O2 or 10% CO2/23% N2/67% O2. The mean control bilateral internal carotid artery blood flow (ICABF) was 23 +/- 10 ml/min (mean +/- SD), mean arterial pressure (MAP) was 101 +/- 13 mm Hg, and PaCO2 was 34 +/- 6 mm Hg. Inhalation of 33% Xe in O2 did not change the ICABF, MAP, or PaCO2. Inhalation of 10% CO2 in O2 increased the ICABF to 39 +/- 15 ml/min (p <0.001), MAP to 112 +/- 16 mm Hg (p <0.05), and PaCO2 to 54 +/- 5 mm Hg (p <0.001). The lack of change in ICABF and PaCO2 with 32% Xe inhalation suggests that a clinically relevant change in CBF is unlikely. PMID- 15815475 TI - The effects of fentanyl and sufentanil on intracranial pressure and cerebral blood flow in rabbits with an acute cryogenic brain injury. AB - The cerebrovascular response to the administration of equipotent doses of fentanyl and sufentanil was evaluated in New Zealand white rabbits following cryogenic brain injury. In a preliminary study consisting of 10 animals, it was documented that the cerebral blood flow response to alterations in the PaCO2 remained intact in this model of brain injury. Subsequently, 28 rabbits were anesthetized with 1.5% halothane in oxygen, paralyzed with pancuronium, and mechanically ventilated. A cryogenic lesion was created over the left hemisphere. One hour later, the intracranial pressure had risen to a mean value of 15 mm Hg. Baseline measurements were then made of monitored variables, which included heart rate, mean arterial pressure, central venous pressure, intracranial pressure, temperature, and arterial blood gases. Global cerebral blood flow was measured utilizing a hydrogen clearance technique. The animals were then randomized to receive an infusion of fentanyl (N = 9, 200 microg/kg), sufentanil (N = 10, 20 microg/kg), or an equal volume of normal saline (N = 9) by i.v. infusion over 5 min. At the conclusion of the opioid infusions, repeated measurements of hemodynamic variables and intracranial pressure were recorded for 15 min and a second cerebral blood flow measurement was made. There were no significant differences in mean arterial pressure, heart rate, central venous pressure, intracranial pressure, cerebral blood flow, or blood gas values between the three groups prior to the administration of fentanyl, sufentanil, or normal saline. At the conclusion of the 5 min infusion, the intracranial pressure had increased by approximately 5 mm Hg in all three groups. The mean arterial pressure decreased to a similar degree in the fentanyl and sufentanil groups and was significantly lower than the mean arterial pressure in the saline group. Although the cerebral perfusion pressure decreased in all three groups, cerebral blood flow was not significantly affected. These results suggest that there is no significant difference in the effects of fentanyl vs. sufentanil on mean arterial pressure, intracranial pressure, or cerebral blood flow in this model of acute brain injury and elevated intracranial pressure. PMID- 15815476 TI - Effect of 80% Xe on whole brain blood flow and metabolism in awake monkeys. AB - We previously reported that 33% xenon (Xe) did not activate cerebral blood flow (CBF) and metabolism in monkeys as it appears to do in humans. However, monkeys may be less sensitive to Xe than humans are, which would explain the discrepancy in the results, but no one has studied the effects of higher concentrations of Xe on CBF and metabolism in monkeys. Therefore, we studied the effect of 80% Xe on whole-brain CBF, cerebral metabolic rate for oxygen (CMRO2) and glucose (CMRG) in five awake rhesus monkeys. Platinum microelectrodes and catheters inserted into the torcular Herophili were used to measure H2 clearance CBF, and to withdraw cerebral venous blood for O2 and glucose analysis. Cerebral variables were measured after 15 min exposure to 80% N2/20% O2 followed by 80% Xe/20% O2. Eighty percent Xe compared with 80% N2 increased (p <0.01) CBF by 52.7% from 74 +/- 16 to 113 +/- 25 (mean +/- SD) ml 100 g(-1)/min(-1), CBF/CMRO by 154% from 13 to 33, and decreased (p <0.05) CMRO2 by 39.3% from 6.1 +/- 0.9 to 3.7 +/- 0.8 ml 100 g( 1)/min(-1) and CMRG by 52.4% from 8.4 +/- 2.6 to 4.0 +/- 2.0 mg 100 g(-1)/min( 1). Electroencephalogram frequency decreased from a predominantly alpha to dagger rhythm in three of five monkeys. The 40 and 50% reduction in CMRO2 and CMRG, respectively, by 80% Xe suggests an anesthetic effect at this dose in the rhesus monkey but also activates CBF by 50%. PMID- 15815478 TI - Use of nitrous oxide in neuroanesthesia: why bother? PMID- 15815479 TI - Place of nitrous oxide in neuroanesthesia: still a valuable drug. PMID- 15815546 TI - [Novel CT examinations using various kinds of image processing]. PMID- 15815547 TI - [Fundamentals in skeletal radiology]. PMID- 15815548 TI - [Imaging techniques and findings of cerebral disease on CT]. PMID- 15815550 TI - [Introduction to medical informatics]. PMID- 15815549 TI - [Thoughts on evidence based medicine for angiography]. PMID- 15815551 TI - [Cardiovascular imaging of MRI: basics and clinical applications]. PMID- 15815552 TI - [Committee report on supertechnologist approval system]. PMID- 15815553 TI - [Report of the Special Committee for the Future Planning on the JSRT (2003)]. PMID- 15815554 TI - [Impressions on the Society Of Nuclear Medicine 51st Annual Meeting]. PMID- 15815555 TI - [Certification seminar for radiotherapy technology specialists]. PMID- 15815556 TI - [MHLW GCP for medical device]. PMID- 15815557 TI - [New method of estimating effective energy for X-ray CT scanners]. AB - Because the exposure dose in X-ray computed tomography examinations is sometimes difficult to determine, it is important to be able to estimate the dose for these examinations. The effective energy of the X-ray CT scanner is required to estimate exposure dose. Although the half-value-layer (HVL) method has been used to calculate effective energy, it is not an easy method. This paper proposes a technique by which effective energy can be easily calculated. Certain details were found to cause change in effective energy, and the ratio (inner-metal center air ratio: IMCAR) between air dose and dose in fixing the metallic pipe in the isocenter of an X-ray CT scanner was necessary. The IMCAR from a different X-ray CT scanner was required, and, when effective energy was calculated, it showed an error of less than 0.7% for the half-value-layer method. The effect of this error on dose estimation was slight (0.4%). This technique is useful, because effective energy can easily be calculated with a high degree of accuracy. PMID- 15815558 TI - [Absorbed dose conversion factors obtained from X-ray spectra measured at water phantom surface]. AB - The absorbed dose conversion factor for X-rays at the water phantom surface has been obtained from the measured spectra. These measurements have been made at tube voltages of 60 kV to 120 kV and field sizes ranging from 5 x 5 cm(2) to 30 x 30 cm(2) with and without additional 2 mm aluminium filtration. A small silicon diode detector with little angular dependence was used for this measurement. The absorbed dose conversion factor obtained was 0.03-0.43% smaller than that obtained from the primary X-ray spectrum. The difference was large for high voltage and heavily filtered X-rays. As field size increases, the conversion factor decreases, but the decrease is slight when field size exceeds 20 x 20 cm(2). The absorbed dose conversion factor obtained from the primary or surface X ray spectrum is 0.4-1.8% larger than that obtained from the effective energy of primary X-rays. The difference is large in high-voltage X-rays and decreases slightly with increases in field size. PMID- 15815560 TI - [Evaluation of time resolution in cardiac synchronized image reconstruction using multi-slice CT]. AB - One of the newest CT application technologies is cardiac synchronized image reconstruction. In this technology, evaluation of time-resolution is very important. We developed a method of measuring time-resolution in cardiac synchronized reconstruction, and evaluated various scanning protocols. In our experiment, ECG-gated scanning was done by multi-slice CT (Aquilion16 Super Heart Edition, Toshiba Medical Systems Co., Ltd., Japan). The nominal slice thickness was 0.5 mm, and rotation time was 0.5 sec. Input heart rate was set at 40, 45, 50, 55, 60, 70, 75, 80, and 90 bpm, and helical pitch at 3.2, 4.0, and 4.8 (beam pitch: 0.200, 0.250 and 0.300). We measured FWTM of the obtained sensitivity distribution and compared at each scanning protocol. Time resolution improved as helical pitch decreased and heart rate increased. However, phase-time resolution deteriorated as heart rate increased. The results of our experiment indicated that a segment center was determined by X-ray tube rotation time and heart rate, and the number of segments was determined by heart rate, helical pitch, and reconstruction position. Time resolution changed with X-ray tube rotation time, heart rate, helical pitch, and reconstruction position. In this report, we provide a reference for an optimal scanning protocol in cardiac synchronized image reconstruction. PMID- 15815559 TI - [With-time change of hemodynamics in fMRI: linearity consideration after stimulation ends from stimulation beginning]. AB - The BOLD signal responses that occur after a few seconds at the beginning and the end of stimulus are due to hemodynamic delay. A temporary increase of deoxyhemoglobin just after the beginning of stimulus causes an initial dip in the hemodynamic signal (response). After the initial dip, an increase of BOLD signal causes an overshoot. After the overshoot, a decrease of the BOLD signal due to the end of stimulus causes an undershoot. Many reports have examined the period of stimulus, but only a few have described the period after stimulus. This paper reports on a peak between the overshoot and undershoot. After the end of stimulus, the signal does not simply decrease, but shows a peak after some delay. An activated map showed the same tendency as the hemodynamics signal. This peak was termed hemodynamic over-saturation (HOS). Time control in chronological order is important to obtain a hemodynamic change in chronological order and a large, significant difference between the task and rest signal. The entire hemodynamic signal was evaluated by controlling the timing of the scan using an external trigger and by indicating the beginning of the task (rest) period using sound. The timing of the external trigger and sound are built into the sequence design as a program. From the results of the hemodynamic signal, the causal relationship between this signal and the fluctuation in oxyhemoglobin and deoxyhemoglobin can be considered. PMID- 15815561 TI - [Selection of parameters for multi-objective frequency processing in CR mammography: visual evaluation of mammographic phantom images]. AB - The multi-objective frequency processing installed in the FCR5000R (Fuji Film Medical) is superior to the conventional processing used in the FCR9000 (Fuji Film Medical) in evaluating frequency processing. A suitable combination of parameters for multi-frequency-processing in computed radiography (CR) mammography was evaluated. The paired-comparison method using phantom images was performed for the visual evaluation. Results showed that the evaluation score of mass and fiber lesions depended on the multi-frequency balance type (MRB) parameter, and the low-frequency-cycle emphasizing parameter had the highest score. In contrast, the score of microcalcifications depended on the degree of multi-frequency enhancement (MRE). The most suitable parameters for the multi frequency processing of every size and type of breast lesion were not obtained. However, MRB=A, MRT=p, and MRE=1.0 can be recommend for CR mammography. PMID- 15815562 TI - Outbreaks of Salmonella infections associated with eating Roma tomatoes--United States and Canada, 2004. AB - Three outbreaks of Salmonella infections associated with eating Roma tomatoes were detected in the United States and Canada in the summer of 2004. In one multistate U.S. outbreak during June 25-July 18, multiple Salmonella serotypes were isolated, and cases were associated with exposure to Roma tomatoes from multiple locations of a chain delicatessen. Each of the other two outbreaks was characterized by a single Salmonella serotype: Braenderup in one multistate outbreak and Javiana in an outbreak in Canada. In the three outbreaks, 561 outbreak-related illnesses from 18 states and one province in Canada were identified. This report describes the subsequent investigations by public health and food safety agencies. Although a single tomato-packing house in Florida was common to all three outbreaks, other growers or packers also might have supplied contaminated Roma tomatoes that resulted in some of the illnesses. Environmental investigations are continuing. Because current knowledge of mechanisms of tomato contamination and methods of eradication of Salmonella in fruit is inadequate to ensure produce safety, further research should be a priority for the agricultural industry, food safety agencies, and the public health community. PMID- 15815563 TI - Update: influenza activity--United States, 2004-05 season. AB - This report summarizes influenza activity in the United States during October 3, 2004-March 26, 2005, updates the previous summary, and describes the composition of the 2005-06 influenza vaccine. Influenza activity was moderate in the United States overall, but varied by region. Preliminary data collected through the seven components of the CDC Influenza Surveillance indicate that national influenza activity peaked in early-February. PMID- 15815564 TI - Diabetes-related preventive-care practices--Guam, 2001-2003. AB - Persons with diabetes are at risk for serious complications, such as blindness, kidney failure, nontraumatic lower-extremity amputations, and cardiovascular disease. Preventive-care practices have been determined effective in reducing both the incidence and progression of diabetes-specific complications. Despite the benefits of these practices, their level of use has been lower than recommended in the United States. To emphasize the importance of preventive-care practices, national health objectives for 2010 for persons with diabetes, include the following targets: have an annual dilated eye examination (75%; objective 5 13), have an annual foot examination (75%; objective 5-14), perform self monitoring of blood glucose (SMBG) at least once daily (60%; objective 5-17), and have a glycated hemoglobin (HbA1c) measurement at least twice per year (65%; objective 5-12 [revised]). In the U.S. territory of Guam (2004 population: 166,090), no previous population-based assessment of the use of diabetes-related preventive-care practices has been conducted. For this report, data from the 2001 2003 Guam Behavioral Risk Factor Surveillance System (BRFSS) were analyzed to determine the prevalence of preventive-care practices among persons with diabetes in Guam, which is the southernmost and largest of the Marianas Islands, located approximately 3,300 miles west of Hawaii and 1,550 miles south of Japan. Results of the analysis indicated that Guam residents with diabetes remain below the national targets for 2010 for four preventive-care practices, most notably SMBG. The preventive care programs and surveillance activities of the Guam Diabetes Prevention and Control Program (DPCP) should be continued, with emphasis on SMBG recommendations, to prevent poor health outcomes in persons with diabetes and achieve the national health objectives. PMID- 15815566 TI - Effect of educational preparation on the accuracy of linear growth measurement in pediatric primary care practices: results of a multicenter nursing study. AB - Consistently monitoring a child's linear growth is one of the least invasive, most sensitive tools to identify normal physiologic functioning and a healthy lifestyle. However, studies, mostly from the United Kingdom, indicate that children are frequently measured incorrectly. Inaccurate linear measurements may result in some children having undetected growth disorders whereas others with normal growth being referred for costly, unwarranted specialty evaluations. This study presents the secondary analysis of a primary study that used a randomized control study design to demonstrate that a didactic educational intervention resulted in significantly more children being measured accurately within eight pediatric practices. The secondary analysis explored the influence of the measurer's educational level on the outcome of accurate linear measurement. Results indicated that RNs were twice as likely as non-RNs to measure children accurately. PMID- 15815567 TI - Puberty-timing is everything! AB - Puberty is a dynamic period of physical growth, sexual maturation, and psychosocial achievement that generally begins between age 8 and 14 years. The age of onset varies as a function of sex, ethnicity, health status, genetics, nutrition, and activity level. Puberty is initiated by hormonal changes triggered by the hypothalamus. Children with variants of normal pubertal development--both early and late puberty--are common in pediatric practice. Recognizing when variations are normal and when referral for further evaluation is indicated is an important skill. PMID- 15815568 TI - Education and telephone case management for children with type 1 diabetes: a randomized controlled trial. AB - The purpose of the study was to compare three nursing interventions and their impact on glycemic control among children with type I diabetes. The 75 subjects' mean +/- SD age was 12.5 +/- 3.4 years, 55% were boys, and 55% were White. Subjects were randomly assigned to a standard care (SC), an education (ED), or an education and telephone case management (ED + TCM) group. The primary outcome measure was glycemic control (hemoglobin A1c, or HbA1c). Secondary outcome measures were diabetes knowledge (KNOW), parent-child teamwork (TEAM), and adherence (ADH). After 6 months of follow-up, results demonstrated no significant differences among groups in HbA1c. KNOW and TEAM scores improved slightly in the ED and ED + TCM groups, but no statistically significant differences were found among the three groups. Significant improvement in ADH scores among ED + TCM groups was reported when compared with the ED and SC groups. This change may represent a move toward improved adherence to diabetes care and subsequent improvement in diabetes control. The challenges of recruitment and retention of subjects in this study will also be discussed. PMID- 15815569 TI - The care of children and adolescents with type 2 diabetes. AB - The prevalence of Type 2 diabetes has dramatically increased in children and adolescents over the past 10 years. Type 2 diabetes is characterized by insulin resistance and high insulin levels. Reasons cited for the rise of this condition in children and adolescents are speculated to stem from obesity because of a rise in sedentary behavior, nonnutritious food choices, and genetic predisposition. A high recurrence rate in families shows that therapy for children and adolescents must involve the entire family to be successful. Treatment recommendations vary depending on severity but include nutrition, exercise, and medication. Assessment of the patient's and family's willingness to change their current lifestyle behaviors is an integral part of treatment. Nutrition and exercise goals should be made on an individual basis to meet the needs of the patient. Success of therapy is difficult to measure because this is a chronic condition being diagnosed in young people. As in any chronic condition, success of therapy is difficult to measure. PMID- 15815570 TI - Effects of hypoglycemia on developmental outcome in children with congenital hyperinsulinism. AB - Children with congenital hyperinsulinism are at risk for recurring, severe episodes of hypoglycemia that can cause seizures, brain damage, and developmental delay. To assess the frequency of permanent brain damage in this disorder, we carried out a telephone survey of 68 children who presented to The Children's Hospital of Philadelphia between 1980 and 2000. One third of the group had some degree of developmental delay. Those presenting in the first week of life and those with medically unresponsive hyperinsulinism were more likely to have delays. Early recognition and control of hypoglycemia are essential for preventing developmental delay in these children. PMID- 15815571 TI - Advances in assessment, diagnosis, and treatment of hyperthyroidism in children. AB - The thyroid gland is responsible for regulating multiple complex metabolic processes that affect most organs. Physical growth and cognitive development are dependent on proper levels of thyroid hormone. This article will review common challenges in the diagnosis of hyperthyroidism in children, the approaches to treatment, and the nursing interventions guided toward child and family responses to thyroid disease. A comparison of signs and symptoms of hypothyroidism and hyperthyroidism is also included. The nursing interventions addressed in the article integrate the biological, psychological, social, and environmental stresses and adaptations necessary to cope with hyperthyroid disease. PMID- 15815577 TI - Pediatric cardiac transplants: a clinical update. AB - In the last 10 years more than 2,600 pediatric patients have received heart transplants. The survival rate at 10 years is 65% (Blume, 2003). There have been profound improvements in the survival and quality of life associated with pediatric heart transplants. Due to the increase in survival and quality of life, it is becoming more common for nurses to care for these unique patients. Nurses need to be aware of the complex management of pediatric heart transplant patients to aid in their care of these patients. This paper will discuss the transplant process, medications, common complications, and psychosocial impact associated with pediatric heart transplants to better prepare nurses involved in the care of pediatric cardiac transplant patients. PMID- 15815579 TI - Family presence during pediatric resuscitation: an integrative review for evidence-based practice. AB - Many pediatric health care institutions lack guidelines to address the presence of family members during resuscitation activities. This integrative literature review was used to establish an evidence-based approach that would compliment our institution's philosophy of "family centered care." The evidence supports a partnering with the families: Hospital staff ask families whether they want to be present during life-saving interventions by health care professions. Families want to make the choice, and when they are present during resuscitation efforts, there is less litigation, anxiety, and second-guessing regarding the efforts and competence of the staff providing that care. No literature was found to support beliefs that family member presence is harmful to the family or the institution. A practice policy is provided that respects family choice and presence during life-saving measures in pediatric care facilities. PMID- 15815580 TI - Statins and stroke. AB - Inhibitors of HMG-CoA reductase (statins) are potent cholesterol-lowering drugs. Large clinical trials have shown that statins reduce the incidence of cerebrovascular events, which might be surprising because cholesterol is not an established risk factor for stroke. In addition to their cholesterol-lowering properties, statins exert a number of pleiotropic, vasculoprotective actions that include improvement of endothelial function, increased nitric oxide (NO) bioavailability, antioxidant properties, inhibition of inflammatory responses, immunomodulatory actions, regulation of progenitor cells, and stabilization of atherosclerotic plaques. In fact, statins augment cerebral blood flow and confer significant protection in animal models of stroke partly via mechanisms related to the upregulation of endothelial nitric oxide synthase. Retrospective clinical evidence suggests that long-term statin administration may not only reduce stroke risk but also improve outcome. Early secondary prevention trials are underway to test the hypothesis that statin treatment initiated immediately after an event improves short-term outcome. Lastly, recent evidence suggests that sudden discontinuation of statin treatment leads to a rebound effect with downregulation of NO production. Withdrawal of statin treatment may impair vascular function and increase morbidity and mortality in patients with vascular disease. PMID- 15815581 TI - Signaling mechanisms of HIV-1 Tat-induced alterations of claudin-5 expression in brain endothelial cells. AB - Exposure of brain microvascular endothelial cells (BMEC) to human immunodeficiency virus-1 (HIV-1) Tat protein can decrease expression and change distribution of tight junction proteins, including claudin-5. Owing to the importance of claudin-5 in maintaining the blood-brain barrier (BBB) integrity, the present study focused on the regulatory mechanisms of Tat-induced alterations of claudin-5 mRNA and protein levels. Real-time reverse-transcription-polymerase chain reaction revealed that claudin-5 mRNA was markedly diminished in BMEC exposed to Tat. However, U0126 (an inhibitor of mitogen-activated protein kinase kinase1/2, MEK1/2) protected against this effect. In addition, inhibition of the vascular endothelial growth factor receptor type 2 (VEGFR-2) by SU1498, phosphatidylinositol-3 kinase (PI-3 K) by LY294002, nuclear factor-kappaB (NF kappaB) by peptide SN50, and intracellular calcium by BAPTA/AM partially prevented Tat-mediated alterations in claudin-5 protein levels and immunoreactivity patterns. In contrast, inhibition of protein kinase C did not affect claudin-5 expression in Tat-treated cells. The present findings indicate that activation of VEGFR-2 and multiple redox-regulated signal transduction pathways are involved in Tat-induced alterations of claudin-5 expression. Because claudins constitute the major backbone of tight junctions, the present data are relevant to the disturbances of the BBB in the course of HIV-1 infection. PMID- 15815582 TI - Atypical hemispheric dominance for attention: functional MRI topography. AB - The right hemisphere is predominantly involved in tasks associated with spatial attention. However, left hemispheric dominance for spatial attention can be found in healthy individuals, and both spatial attention and language can be lateralized to the same hemisphere. Little is known about the underlying regional distribution of neural activation in these 'atypical' individuals. Previously a large number of healthy subjects were screened for hemispheric dominance of visuospatial attention and language, using functional Doppler ultrasonography. From this group, subjects were chosen who were 'atypical' for hemispheric dominance of visuospatial attention and language, and their pattern of brain activation was studied with functional magnetic resonance imaging during a task probing spatial attention. Right-handed subjects with the 'typical' pattern of brain organization served as control subjects. It was found that subjects with an inverted lateralization of language and spatial attention (language right, attention left) recruited left-hemispheric areas in the attention task, homotopic to those recruited by control subjects in the right hemisphere. Subjects with lateralization of both language and attention to the right hemisphere activated an attentional network in the right hemisphere that was comparable to control subjects. The present findings suggest that not the hemispheric side, but the intrahemispheric pattern of activation is the distinct feature for the neural processes underlying language and attention. PMID- 15815583 TI - Cerebral metabolic response to low blood flow: possible role of cytochrome oxidase inhibition. AB - The reactions of cerebral metabolism to imposed changes of cerebral blood flow (CBF) are poorly understood. A common explanation of the mismatched CBF and oxygen consumption (CMR(O(2))) during neuronal excitation holds that blood flow rises more than oxygen consumption to compensate for an absent oxygen reserve in brain mitochondria. The claim conversely implies that oxygen consumption must decline when blood flow declines. As the prevailing rate of reaction of oxygen with cytochrome c oxidase is linked to the tension of oxygen, the claim fails to explain how oxygen consumption is maintained during moderate reductions of CBF imposed by hyperventilation (hypocapnia) or cyclooxygenase (COX) inhibition. To resolve this contradiction, we extended the previously published oxygen delivery model with a term allowing for the adjustment of the affinity of cytochrome c oxidase to a prevailing oxygen tension. The extended model predicted constant oxygen consumption at moderately reduced blood flow. We determined the change of affinity of cytochrome c oxidase in the extended model by measuring CBF in seven, and CMR(O(2)) in five, young healthy volunteers before and during COX inhibition with indomethacin. The average CBF declined 35%, while neither regional nor average CMR(O(2)) changed significantly. The adjustment of cytochrome c oxidase affinity to the declining oxygen delivery could be ascribed to a hypothetical factor with several properties in common with nitric oxide. PMID- 15815584 TI - Activation of the PI3-K/Akt pathway mediates cGMP enhanced-neurogenesis in the adult progenitor cells derived from the subventricular zone. AB - The intracellular mechanisms that regulate neurogenesis remain unclear. Using neurospheres isolated from the subventricular zone (SVZ) of the adult rat, we investigated the effect of cyclic guanosine monophosphate (cGMP) and its signaling pathway on the induction of neurogenesis. Neurospheres expressed phosphodiesterase 5 (PDE5) and treatment of neurospheres with Sildenafil, a specific inhibitor of PDE5, significantly increased cGMP levels and neurogenesis. In addition, incubation of neurospheres with Sildenafil significantly phosphorylated Akt, which was associated with an increase of phosphorylation of glycogen synthase kinase 3 (GSK-3), a downstream target of Akt. Coincubation of neurospheres with Sildenafil and LY 294002, a pharmacological inhibitor of PI3 K/Akt, abolished Sildenafil-induced phosphorylated Akt and GSK-3. Furthermore, LY 294002 blocked Sildenafil-increased SVZ cell proliferation. These data suggest that Sildenafil-enhanced neurogenesis likely occurs through activation of the PI3 K/Akt/GSK-3 pathway. PMID- 15815585 TI - Endothelin-1 overexpression leads to further water accumulation and brain edema after middle cerebral artery occlusion via aquaporin 4 expression in astrocytic end-feet. AB - Stroke patients have increased levels of endothelin-1 (ET-1), a strong vasoconstrictor, in their plasma or cerebrospinal fluid. Previously, we showed high level of ET-1 mRNA expression in astrocytes after hypoxia/ischemia. It is unclear whether the contribution of ET-1 induction in astrocytes is protective or destructive in cerebral ischemia. Here, we generated a transgenic mouse model that overexpress ET-1 in astrocytes (GET-1) using the glial fibrillary acidic protein promoter to examine the role of astrocytic ET-1 in ischemic stroke by challenging these mice with transient middle cerebral artery occlusion (MCAO). Under normal condition, GET-1 mice showed no abnormality in brain morphology, cerebrovasculature, absolute cerebral blood flow, blood-brain barrier (BBB) integrity, and mean arterial blood pressure. Yet, GET-1 mice subjected to transient MCAO showed more severe neurologic deficits and increased infarct, which were partially normalized by administration of ABT-627 (ET(A) antagonist) 5 mins after MCAO. In addition, GET-1 brains exhibited more Evans blue extravasation and showed decreased endothelial occludin expression after MCAO, correlating with higher brain water content and increased cerebral edema. Aquaporin 4 expression was also more pronounced in astrocytic end-feet on blood vessels in GET-1 ipsilateral brains. Our current data suggest that astrocytic ET 1 has deleterious effects on water homeostasis, cerebral edema and BBB integrity, which contribute to more severe ischemic brain injury. PMID- 15815587 TI - Release of bradykinin and expression of kinin B2 receptors in the brain: role for cell death and brain edema formation after focal cerebral ischemia in mice. AB - Pharmacological studies using bradykinin B2 receptor antagonists suggest that bradykinin, an early mediator of inflammation and the main metabolite of the kallikrein-kinin system, is involved in secondary brain damage after cerebral ischemia. However, the time-course of bradykinin production and kinin receptor expression as well as the conclusive role of bradykinin B2 receptors for brain damage after experimental stroke have not been elucidated so far. C57/Bl6 mice were subjected to 45 mins of middle cerebral artery occlusion (MCAO) and 2, 4, 8, 24, and 48 h later brains were removed for the analysis of tissue bradykinin concentration and kinin B2 receptor mRNA and protein expression. Brain edema, infarct volume, functional outcome, and long-term survival were assessed in WT and B2-/- mice 24 h or 7 days after MCAO. Tissue bradykinin was maximally increased 12 h after ischemia (three-fold), while kinin B2 receptor mRNA upregulation peaked 24 to 48 h after MCAO (10- to 12-fold versus naive brain tissue). Immunohistochemistry revealed that kinin B2 receptors were constitutively and widely expressed in mouse brain, were upregulated 2 h after ischemia in cells showing signs of ischemic damage, and remained upregulated in the penumbra up to 24 h after ischemia. B2-/- mice had improved motor function (P<0.05), smaller infarct volumes (-38%; P<0.01), developed less brain edema ( 87%; P<0.05), and survived longer (P<0.01) as compared with wild-type controls. The current results show that bradykinin is produced in the brain, kinin B2 receptors are upregulated on dying cells, and B2 receptors are involved in cell death and brain edema formation after experimental stroke. PMID- 15815588 TI - Uncomfortable truths. PMID- 15815586 TI - Expression of cellular FLICE inhibitory proteins (cFLIP) in normal and traumatic murine and human cerebral cortex. AB - Cellular Fas-associated death domain-like interleukin-1-beta converting enzyme (FLICE) inhibitory proteins (cFLIPs) are endogenous caspase homologues that inhibit programmed cell death. We hypothesized that cFLIPs are differentially expressed in response to traumatic brain injury (TBI). cFLIP-alpha and cFLIP delta mRNA were expressed in normal mouse brain-specifically cFLIP-delta (but not cFLIP-alpha) protein was robustly expressed. After controlled cortical impact (CCI), cFLIP-alpha expression increased initially then decreased to control levels at 12 h, increasing again at 24-72 h (P<0.05). cFLIP-delta expression was decreased in brain homogenates by 12 h after CCI, then increased again at 24 to 72 h (P<0.05). cFLIP-delta immunostaining was markedly reduced in injured cortex, but not hippocampus, at 3 to 72 h after CCI. In cortex, reduced cFLIP-delta staining was found in TUNEL-positive cells, but in hippocampus TUNEL-positive cells expressed cFLIP-delta immunoreactivity. cFLIP-delta was increased in a subset of reactive astrocytes in pericontusional cortex and hippocampus at 48 to 72 h. Low levels of both cFLIP isoforms were detected in human cortical tissue with no TBI, from four patients undergoing brain surgery for epilepsy and <24 h post mortem from three patients without CNS pathologic assessment. In cortical tissue surgically removed <18 h after severe TBI (n=3), cFLIP-alpha expression was increased relative to epilepsy controls (P<0.05) but not relative to post mortem controls. The data suggest differential spatial and temporal regulation of cFLIP-alpha and cFLIP-delta expression that may influence the magnitude of cell death and further implicate programmed mechanisms of cell death after TBI. PMID- 15815590 TI - Nuclear chiefs scotch story on frailty of ageing warheads. PMID- 15815589 TI - Warning system steps up a gear for fresh Indonesian earthquake. PMID- 15815591 TI - Pope praised for partial conciliation of science and religion. PMID- 15815592 TI - Image problems jeopardize comet mission's impact. PMID- 15815593 TI - Job switch stymies Japan's abduction probe. PMID- 15815594 TI - Professors bristle as states act to mould lecture content. PMID- 15815595 TI - Global health agency split over potential anti-terrorism duties. PMID- 15815596 TI - Postdocs slam zealous attitude of NIH ethics office. PMID- 15815597 TI - US abandons health study on Agent Orange. PMID- 15815599 TI - Tropical medicine: melioidosis? Never heard of it... PMID- 15815600 TI - Stem-cell research: after the gold rush. PMID- 15815601 TI - Don't mix radiocarbon and calendar years. PMID- 15815602 TI - DNA barcoding is no substitute for taxonomy. PMID- 15815603 TI - Arrogance imperils plans for change at Harvard. PMID- 15815604 TI - A global call for new polio vaccines. PMID- 15815609 TI - Stirring the primordial soup. PMID- 15815610 TI - Planetary science: a planet that blinks. PMID- 15815611 TI - Pharmacology: marijuana and your heart. PMID- 15815612 TI - Tuberculosis: the genetics of vulnerability. PMID- 15815614 TI - Technology: hydrogen and hydrates. PMID- 15815615 TI - Environmental science: germ theory for ailing corals. PMID- 15815616 TI - Developmental biology: reproduction in clusters. PMID- 15815617 TI - Evolutionary biology: channels of resistance. PMID- 15815618 TI - Anthropology: the earliest toothless hominin skull. AB - The site of Dmanisi in the Eurasian republic of Georgia has yielded striking hominin, faunal and archaeological material as evidence for the presence of early Homo outside Africa 1.77 million years ago, documenting an important episode in human evolution. Here we describe a beautifully preserved skull and jawbone from a Dmanisi hominin of this period who had lost all but one tooth several years before death. This specimen not only represents the earliest case of severe masticatory impairment in the hominin fossil record to be discovered so far, but also raises questions about alternative subsistence strategies in early Homo. PMID- 15815619 TI - History of science: Dante's insight into galilean invariance. AB - In 1632, Galileo described his experience of motion aboard a large ship and exposed in detail the invariance principle, which was then rightly named after him. I suggest that more than three centuries earlier, in The Divine Comedy, his fellow countryman Dante Alighieri intuitively grasped what Galileo was later to establish as one of the pillars of modern science. PMID- 15815620 TI - Modes of faulting at mid-ocean ridges. AB - Abyssal-hill-bounding faults that pervade the oceanic crust are the most common tectonic feature on the surface of the Earth. The recognition that these faults form at plate spreading centres came with the plate tectonic revolution. Recent observations reveal a large range of fault sizes and orientations; numerical models of plate separation, dyke intrusion and faulting require at least two distinct mechanisms of fault formation at ridges to explain these observations. Plate unbending with distance from the top of an axial high reproduces the observed dip directions and offsets of faults formed at fast-spreading centres. Conversely, plate stretching, with differing amounts of constant-rate magmatic dyke intrusion, can explain the great variety of fault offset seen at slow spreading ridges. Very-large-offset normal faults only form when about half the plate separation at a ridge is accommodated by dyke intrusion. PMID- 15815622 TI - Recruitment of entomopathogenic nematodes by insect-damaged maize roots. AB - Plants under attack by arthropod herbivores often emit volatile compounds from their leaves that attract natural enemies of the herbivores. Here we report the first identification of an insect-induced belowground plant signal, (E)-beta caryophyllene, which strongly attracts an entomopathogenic nematode. Maize roots release this sesquiterpene in response to feeding by larvae of the beetle Diabrotica virgifera virgifera, a maize pest that is currently invading Europe. Most North American maize lines do not release (E)-beta-caryophyllene, whereas European lines and the wild maize ancestor, teosinte, readily do so in response to D. v. virgifera attack. This difference was consistent with striking differences in the attractiveness of representative lines in the laboratory. Field experiments showed a fivefold higher nematode infection rate of D. v. virgifera larvae on a maize variety that produces the signal than on a variety that does not, whereas spiking the soil near the latter variety with authentic (E)-beta-caryophyllene decreased the emergence of adult D. v. virgifera to less than half. North American maize lines must have lost the signal during the breeding process. Development of new varieties that release the attractant in adequate amounts should help enhance the efficacy of nematodes as biological control agents against root pests like D. v. virgifera. PMID- 15815623 TI - Rapid growth of black holes in massive star-forming galaxies. AB - The tight relationship between the masses of black holes and galaxy spheroids in nearby galaxies implies a causal connection between the growth of these two components. Optically luminous quasars host the most prodigious accreting black holes in the Universe, and can account for greater than or approximately equal to 30 per cent of the total cosmological black-hole growth. As typical quasars are not, however, undergoing intense star formation and already host massive black holes (> 10(8)M(o), where M(o) is the solar mass), there must have been an earlier pre-quasar phase when these black holes grew (mass range approximately (10(6)-10(8))M(o)). The likely signature of this earlier stage is simultaneous black-hole growth and star formation in distant (redshift z > 1; >8 billion light years away) luminous galaxies. Here we report ultra-deep X-ray observations of distant star-forming galaxies that are bright at submillimetre wavelengths. We find that the black holes in these galaxies are growing almost continuously throughout periods of intense star formation. This activity appears to be more tightly associated with these galaxies than any other coeval galaxy populations. We show that the black-hole growth from these galaxies is consistent with that expected for the pre-quasar phase. PMID- 15815624 TI - Tuning clathrate hydrates for hydrogen storage. AB - The storage of large quantities of hydrogen at safe pressures is a key factor in establishing a hydrogen-based economy. Previous strategies--where hydrogen has been bound chemically, adsorbed in materials with permanent void space or stored in hybrid materials that combine these elements--have problems arising from either technical considerations or materials cost. A recently reported clathrate hydrate of hydrogen exhibiting two different-sized cages does seem to meet the necessary storage requirements; however, the extreme pressures (approximately 2 kbar) required to produce the material make it impractical. The synthesis pressure can be decreased by filling the larger cavity with tetrahydrofuran (THF) to stabilize the material, but the potential storage capacity of the material is compromised with this approach. Here we report that hydrogen storage capacities in THF-containing binary-clathrate hydrates can be increased to approximately 4 wt% at modest pressures by tuning their composition to allow the hydrogen guests to enter both the larger and the smaller cages, while retaining low-pressure stability. The tuning mechanism is quite general and convenient, using water soluble hydrate promoters and various small gaseous guests. PMID- 15815621 TI - Generation and annotation of the DNA sequences of human chromosomes 2 and 4. AB - Human chromosome 2 is unique to the human lineage in being the product of a head to-head fusion of two intermediate-sized ancestral chromosomes. Chromosome 4 has received attention primarily related to the search for the Huntington's disease gene, but also for genes associated with Wolf-Hirschhorn syndrome, polycystic kidney disease and a form of muscular dystrophy. Here we present approximately 237 million base pairs of sequence for chromosome 2, and 186 million base pairs for chromosome 4, representing more than 99.6% of their euchromatic sequences. Our initial analyses have identified 1,346 protein-coding genes and 1,239 pseudogenes on chromosome 2, and 796 protein-coding genes and 778 pseudogenes on chromosome 4. Extensive analyses confirm the underlying construction of the sequence, and expand our understanding of the structure and evolution of mammalian chromosomes, including gene deserts, segmental duplications and highly variant regions. PMID- 15815625 TI - Water content in the transition zone from electrical conductivity of wadsleyite and ringwoodite. AB - The distribution of water in the Earth's interior reflects the way in which the Earth has evolved, and has an important influence on its material properties. Minerals in the transition zone of the Earth's mantle (from approximately 410 to approximately 660 km depth) have large water solubility, and hence it is thought that the transition zone might act as a water reservoir. When the water content of the transition zone exceeds a critical value, upwelling flow might result in partial melting at approximately 410 km, which would affect the distribution of certain elements in the Earth. However, the amount of water in the transition zone has remained unknown. Here we determined the effects of water and temperature on the electrical conductivity of the minerals wadsleyite and ringwoodite to infer the water content of the transition zone. We find that the electrical conductivity of these minerals depends strongly on water content but only weakly on temperature. By comparing these results with geophysically inferred conductivity, we infer that the water content in the mantle transition zone varies regionally, but that its value in the Pacific is estimated to be approximately 0.1-0.2 wt%. These values significantly exceed the estimated critical water content in the upper mantle, suggesting that partial melting may indeed occur at approximately 410 km depth, at least in this region. PMID- 15815626 TI - Particle size and energetics of gouge from earthquake rupture zones. AB - Grain size reduction and gouge formation are found to be ubiquitous in brittle faults at all scales, and most slip along mature faults is observed to have been localized within gouge zones. This fine-grain gouge is thought to control earthquake instability, and thus understanding its properties is central to an understanding of the earthquake process. Here we show that gouge from the San Andreas fault, California, with approximately 160 km slip, and the rupture zone of a recent earthquake in a South African mine with only approximately 0.4 m slip, display similar characteristics, in that ultrafine grains approach the nanometre scale, gouge surface areas approach 80 m2 g(-1), and grain size distribution is non-fractal. These observations challenge the common perception that gouge texture is fractal and that gouge surface energy is a negligible contributor to the earthquake energy budget. We propose that the observed fine grain gouge is not related to quasi-static cumulative slip, but is instead formed by dynamic rock pulverization during the propagation of a single earthquake. PMID- 15815627 TI - New material of the earliest hominid from the Upper Miocene of Chad. AB - Discoveries in Chad by the Mission Paleoanthropologique Franco-Tchadienne have substantially changed our understanding of early human evolution in Africa. In particular, the TM 266 locality in the Toros-Menalla fossiliferous area yielded a nearly complete cranium (TM 266-01-60-1), a mandible, and several isolated teeth assigned to Sahelanthropus tchadensis and biochronologically dated to the late Miocene epoch (about 7 million years ago). Despite the relative completeness of the TM 266 cranium, there has been some controversy about its morphology and its status in the hominid clade. Here we describe new dental and mandibular specimens from three Toros-Menalla (Chad) fossiliferous localities (TM 247, TM 266 and TM 292) of the same age. This new material, including a lower canine consistent with a non-honing C/P3 complex, post-canine teeth with primitive root morphology and intermediate radial enamel thickness, is attributed to S. tchadensis. It expands the hypodigm of the species and provides additional anatomical characters that confirm the morphological differences between S. tchadensis and African apes. S. tchadensis presents several key derived features consistent with its position in the hominid clade close to the last common ancestor of chimpanzees and humans. PMID- 15815628 TI - Virtual cranial reconstruction of Sahelanthropus tchadensis. AB - Previous research in Chad at the Toros-Menalla 266 fossiliferous locality (about 7 million years old) uncovered a nearly complete cranium (TM 266-01-60-1), three mandibular fragments and several isolated teeth attributed to Sahelanthropus tchadensis. Of this material, the cranium is especially important for testing hypotheses about the systematics and behavioural characteristics of this species, but is partly distorted from fracturing, displacement and plastic deformation. Here we present a detailed virtual reconstruction of the TM 266 cranium that corrects these distortions. The reconstruction confirms that S. tchadensis is a hominid and is not more closely related to the African great apes. Analysis of the basicranium further indicates that S. tchadensis might have been an upright biped, suggesting that bipedalism was present in the earliest known hominids, and probably arose soon after the divergence of the chimpanzee and human lineages. PMID- 15815629 TI - Evolutionary diversification of TTX-resistant sodium channels in a predator-prey interaction. AB - Understanding the molecular genetic basis of adaptations provides incomparable insight into the genetic mechanisms by which evolutionary diversification takes place. Whether the evolution of common traits in different lineages proceeds by similar or unique mutations, and the degree to which phenotypic evolution is controlled by changes in gene regulation as opposed to gene function, are fundamental questions in evolutionary biology that require such an understanding of genetic mechanisms. Here we identify novel changes in the molecular structure of a sodium channel expressed in snake skeletal muscle, tsNa(V)1.4, that are responsible for differences in tetrodotoxin (TTX) resistance among garter snake populations coevolving with toxic newts. By the functional expression of tsNa(V)1.4, we show how differences in the amino-acid sequence of the channel affect TTX binding and impart different levels of resistance in four snake populations. These results indicate that the evolution of a physiological trait has occurred through a series of unique functional changes in a gene that is otherwise highly conserved among vertebrates. PMID- 15815630 TI - Sodium channel mutation leading to saxitoxin resistance in clams increases risk of PSP. AB - Bivalve molluscs, the primary vectors of paralytic shellfish poisoning (PSP) in humans, show marked inter-species variation in their capacity to accumulate PSP toxins (PSTs) which has a neural basis. PSTs cause human fatalities by blocking sodium conductance in nerve fibres. Here we identify a molecular basis for inter population variation in PSP resistance within a species, consistent with genetic adaptation to PSTs. Softshell clams (Mya arenaria) from areas exposed to 'red tides' are more resistant to PSTs, as demonstrated by whole-nerve assays, and accumulate toxins at greater rates than sensitive clams from unexposed areas. PSTs lead to selective mortality of sensitive clams. Resistance is caused by natural mutation of a single amino acid residue, which causes a 1,000-fold decrease in affinity at the saxitoxin-binding site in the sodium channel pore of resistant, but not sensitive, clams. Thus PSTs might act as potent natural selection agents, leading to greater toxin resistance in clam populations and increased risk of PSP in humans. Furthermore, global expansion of PSP to previously unaffected coastal areas might result in long-term changes to communities and ecosystems. PMID- 15815631 TI - Ipr1 gene mediates innate immunity to tuberculosis. AB - An estimated eight million people are infected each year with the pathogen Mycobacterium tuberculosis, and more than two million die annually. Yet only about 10% of those infected develop tuberculosis. Genetic variation within host populations is known to be significant in humans and animals, but the nature of genetic control of host resistance to tuberculosis remains poorly understood. Previously we mapped a new genetic locus on mouse chromosome 1, designated sst1 (for supersusceptibility to tuberculosis 1). Here we show that this locus mediates innate immunity in sst1 congenic mouse strains and identify a candidate gene, Intracellular pathogen resistance 1 (Ipr1), within the sst1 locus. The Ipr1 gene is upregulated in the sst1 resistant macrophages after activation and infection, but it is not expressed in the sst1 susceptible macrophages. Expression of the Ipr1 transgene in the sst1 susceptible macrophages limits the multiplication not only of M. tuberculosis but also of Listeria monocytogenes and switches a cell death pathway of the infected macrophages from necrosis to apoptosis. Our data indicate that the Ipr1 gene product might have a previously undocumented function in integrating signals generated by intracellular pathogens with mechanisms controlling innate immunity, cell death and pathogenesis. PMID- 15815632 TI - Low dose oral cannabinoid therapy reduces progression of atherosclerosis in mice. AB - Atherosclerosis is a chronic inflammatory disease, and is the primary cause of heart disease and stroke in Western countries. Derivatives of cannabinoids such as delta-9-tetrahydrocannabinol (THC) modulate immune functions and therefore have potential for the treatment of inflammatory diseases. We investigated the effects of THC in a murine model of established atherosclerosis. Oral administration of THC (1 mg kg(-1) per day) resulted in significant inhibition of disease progression. This effective dose is lower than the dose usually associated with psychotropic effects of THC. Furthermore, we detected the CB2 receptor (the main cannabinoid receptor expressed on immune cells) in both human and mouse atherosclerotic plaques. Lymphoid cells isolated from THC-treated mice showed diminished proliferation capacity and decreased interferon-gamma secretion. Macrophage chemotaxis, which is a crucial step for the development of atherosclerosis, was also inhibited in vitro by THC. All these effects were completely blocked by a specific CB2 receptor antagonist. Our data demonstrate that oral treatment with a low dose of THC inhibits atherosclerosis progression in the apolipoprotein E knockout mouse model, through pleiotropic immunomodulatory effects on lymphoid and myeloid cells. Thus, THC or cannabinoids with activity at the CB2 receptor may be valuable targets for treating atherosclerosis. PMID- 15815633 TI - Impairment of angiogenesis and cell migration by targeted aquaporin-1 gene disruption. AB - Aquaporin-1 (AQP1) is a water channel protein expressed widely in vascular endothelia, where it increases cell membrane water permeability. The role of AQP1 in endothelial cell function is unknown. Here we show remarkably impaired tumour growth in AQP1-null mice after subcutaneous or intracranial tumour cell implantation, with reduced tumour vascularity and extensive necrosis. A new mechanism for the impaired angiogenesis was established from cell culture studies. Although adhesion and proliferation were similar in primary cultures of aortic endothelia from wild-type and from AQP1-null mice, cell migration was greatly impaired in AQP1-deficient cells, with abnormal vessel formation in vitro. Stable transfection of non-endothelial cells with AQP1 or with a structurally different water-selective transporter (AQP4) accelerated cell migration and wound healing in vitro. Motile AQP1-expressing cells had prominent membrane ruffles at the leading edge with polarization of AQP1 protein to lamellipodia, where rapid water fluxes occur. Our findings support a fundamental role of water channels in cell migration, which is central to diverse biological phenomena including angiogenesis, wound healing, tumour spread and organ regeneration. PMID- 15815637 TI - Protein purification: pure but not simple. PMID- 15815641 TI - Guiding hands. PMID- 15815642 TI - Model mentors. PMID- 15815644 TI - Nuts and bolts. International ventures. PMID- 15815648 TI - Reburnishing golden rice. PMID- 15815647 TI - Spatiotemporal regulation of MyD88-IRF-7 signalling for robust type-I interferon induction. AB - Robust type-I interferon (IFN-alpha/beta) induction in plasmacytoid dendritic cells, through the activation of Toll-like receptor 9 (TLR9), constitutes a critical aspect of immunity. It is absolutely dependent on the transcription factor IRF-7, which interacts with and is activated by the adaptor MyD88. How plasmacytoid dendritic cells, but not other cell types (such as conventional dendritic cells), are able to activate the MyD88-IRF-7-dependent IFN induction pathway remains unknown. Here we show that the spatiotemporal regulation of MyD88 IRF-7 signalling is critical for a high-level IFN induction in response to TLR9 activation. The IFN-inducing TLR9 ligand, A/D-type CpG oligodeoxynucleotide (CpG A), is retained for long periods in the endosomal vesicles of plasmacytoid dendritic cells, together with the MyD88-IRF-7 complex. However, in conventional dendritic cells, CpG-A is rapidly transferred to lysosomal vesicles. We further show that conventional dendritic cells can also mount a robust IFN induction if CpG-A is manipulated for endosomal retention using a cationic lipid. This strategy also allows us to demonstrate endosomal activation of the IFN pathway by the otherwise inactive TLR9 ligand B/K-type oligodeoxynucleotide (CpG-B). Thus, our study offers insights into the regulation of TLR9 signalling in space, potentially suggesting a new avenue for therapeutic intervention. PMID- 15815649 TI - Tysabri raises alarm bells on drug class. PMID- 15815650 TI - Peptide leads new class of chronic pain drugs. PMID- 15815651 TI - Syrrx acquisition signals maturation of structure-based discovery. PMID- 15815652 TI - Australian biotech suffers growing pains. PMID- 15815653 TI - Monsanto branches out into fruits and vegetables. PMID- 15815655 TI - Profile: Lester Crawford. PMID- 15815656 TI - Trends in the biotech literature. PMID- 15815657 TI - New apoptosis drugs face critical test. PMID- 15815659 TI - Regulatory frameworks in developing countries. PMID- 15815661 TI - The new fundamentalism. PMID- 15815660 TI - Refuges in India and delayed resistance to Bt crops. PMID- 15815662 TI - Biotech venture capital--it's not too late to be early. PMID- 15815663 TI - Diversify to multiply. PMID- 15815665 TI - Getting real about valuations in biotech. PMID- 15815666 TI - Golden Rice gets a boost from maize. PMID- 15815667 TI - Tackling mastitis in dairy cows. PMID- 15815668 TI - Eco-friendly insect management. PMID- 15815669 TI - Serum of youth? PMID- 15815671 TI - Regulating transgenic crops sensibly: lessons from plant breeding, biotechnology and genomics. AB - The costs of meeting regulatory requirements and market restrictions guided by regulatory criteria are substantial impediments to the commercialization of transgenic crops. Although a cautious approach may have been prudent initially, we argue that some regulatory requirements can now be modified to reduce costs and uncertainty without compromising safety. Long-accepted plant breeding methods for incorporating new diversity into crop varieties, experience from two decades of research on and commercialization of transgenic crops, and expanding knowledge of plant genome structure and dynamics all indicate that if a gene or trait is safe, the genetic engineering process itself presents little potential for unexpected consequences that would not be identified or eliminated in the variety development process before commercialization. We propose that as in conventional breeding, regulatory emphasis should be on phenotypic rather than genomic characteristics once a gene or trait has been shown to be safe. PMID- 15815677 TI - Recent patent applications in RNA interference. PMID- 15815676 TI - Material transfer agreements: open science vs. proprietary claims. PMID- 15815679 TI - A Turing-like test for biological modeling. PMID- 15815680 TI - Alternative career options in the biomedical industry: the professional science master's degree. PMID- 15815682 TI - Gene therapy progress and prospects: targeted gene repair. AB - The capacity to correct a mutant gene within the context of the chromosome holds great promise as a therapy for inherited disorders but fulfilling this promise has proven to be challenging. However, steady progress is being made and the development of gene repair as a viable and robust approach is underway. Here, we present some of the recent advances that are helping to shape our thinking about the feasibility and the limitations of this technique. For the most part, these advances center on understanding the regulation of the reaction and validating its application in animal models. PMID- 15815683 TI - Fyn/Yes and non-canonical Wnt signalling converge on RhoA in vertebrate gastrulation cell movements. AB - Convergent extension (CE) cell movements during gastrulation mediate extension of the anterior-posterior body axis of vertebrate embryos. Non-canonical Wnt5 and Wnt11 signalling is essential for normal CE movements in vertebrate gastrulation. Here, we show that morpholino (MO)-mediated double knock-down of the Fyn and Yes tyrosine kinases in zebrafish embryos impaired normal CE cell movements, resembling the silberblick and pipetail mutants, caused by mutations in wnt11 and wnt5, respectively. Co-injection of Fyn/Yes- and Wnt11- or Wnt5-MO was synergistic, but wnt11 or wnt5 RNA did not rescue the Fyn/Yes knockdown or vice versa. Remarkably, active RhoA rescued the Fyn/Yes knockdown as well as the Wnt11 knockdown, indicating that Fyn/Yes and Wnt11 signalling converged on RhoA. Our results show that Fyn and Yes act together with non-canonical Wnt signalling via RhoA in CE cell movements during gastrulation. PMID- 15815684 TI - FtsY, the bacterial signal-recognition particle receptor, interacts functionally and physically with the SecYEG translocon. AB - Co-translational membrane targeting of proteins by the bacterial signal recognition particle (SRP) requires the specific interaction of the SRP-ribosome nascent chain complex with FtsY, the bacterial SRP receptor (SR). FtsY is homologous to the SRalpha-subunit of the eukaryotic SR, which is tethered to the endoplasmic-reticulum membrane by its interaction with the integral SRbeta subunit. In contrast to SRalpha, FtsY is partly membrane associated and partly located in the cytosol. However, the mechanisms by which FtsY associates with the membrane are unclear. No gene encoding an SRbeta homologue has been found in bacterial genomes, and the presence of an FtsY-specific membrane receptor has not been shown so far. We now provide evidence for the direct interaction between FtsY and the SecY translocon. This interaction offers an explanation of how the bacterial SRP cycle is regulated in response to available translocation channels. PMID- 15815685 TI - Linkage analysis of the genetic determinants of T-cell IL-4 secretion, and identification of Flj20274 as a putative candidate gene. AB - The activation-induced differentiation of naive CD4+ T cells generates functionally divergent type 1 helper T cells (Th1) or type 2 helper T cells (Th2) effector cell populations, characterized by secretion of Interferon (IFN)-gamma or Interleukin (IL)-4, respectively. Inappropriate generation of Th subsets may contribute to immune dysfunction. The decision to generate Th1/Th2 lineages is critically regulated by cytokines, such that IL-12 induces Th1 differentiation, while IL-4 induces Th2 differentiation. Genetic factors influence the pathway of Th differentiation, as displayed by the preferential generation of divergent Th populations by different inbred strains of mice. We employ two complementary genetic techniques to identify genes that regulate the default IL-4 secretion profiles of T cells from BALB/c and B6 mice. We performed a genome-wide linkage analysis of the progeny of a backcross between BALB/c and B6 mice to identify three loci, T-cell secretion of interleukin-4 (Tsi)1-3, on chromosomes 7, 19 and 15, respectively, which regulate in vitro T-cell IL-4 production. We have also employed mRNA representational difference analysis to isolate a gene, Flj20274, which is differentially expressed in T cells that secrete high levels of IL-4. Significantly, Flj20274 was mapped to the point of peak linkage within Tsi1 and is a strong candidate for Tsi1. PMID- 15815686 TI - Promoter region polymorphism of macrophage migration inhibitory factor is strong risk factor for young onset of extensive alopecia areata. AB - We have demonstrated that serum macrophage migration inhibitory factor (MIF) was significantly elevated in patients with extensive alopecia areata (AA). Recently, functional polymorphisms have been identified in the MIF promoter region. To address the functional and prognostic relevance of the -173G/C and -794[CATT]5-8 repeat polymorphisms in MIF genes in patients with extensive AA, 113 patients with extensive AA and 194 healthy controls were genotyped. We found that MIF 173*C was a risk factor for early onset (<20 years) of extensive AA (odds ratio for GC heterozygotes with -173G/C was 4.88 (95% CI, 2.04-11.8), P=0.00038; odds ratio for CC homozygotes with -173G/C was 10.42 (95% CI, 2.56-43.5), P=0.0011). We found no statistically significant differences in the genotype frequencies of the -794[CATT]5-8 repeat polymorphism and extensive AA. These results suggest that polymorphisms within the MIF-173*C allele confer an increased risk of susceptibility to the extensive forms of AA, especially with an early onset of disease. MIF is therefore suggested to be closely implicated in the pathogenesis of the more extensive forms of AA. PMID- 15815687 TI - Real-time RT-PCR normalisation; strategies and considerations. AB - Real-time RT-PCR has become a common technique, no longer limited to specialist core facilities. It is in many cases the only method for measuring mRNA levels of vivo low copy number targets of interest for which alternative assays either do not exist or lack the required sensitivity. Benefits of this procedure over conventional methods for measuring RNA include its sensitivity, large dynamic range, the potential for high throughout as well as accurate quantification. To achieve this, however, appropriate normalisation strategies are required to control for experimental error introduced during the multistage process required to extract and process the RNA. There are many strategies that can be chosen; these include normalisation to sample size, total RNA and the popular practice of measuring an internal reference or housekeeping gene. However, these methods are frequently applied without appropriate validation. In this review we discuss the relative merits of different normalisation strategies and suggest a method of validation that will enable the measurement of biologically meaningful results. PMID- 15815688 TI - Association of interferon gamma and interleukin 10 genes with tuberculosis in Hong Kong Chinese. AB - Interferon gamma (IFN-gamma) and interleukin 10 (IL-10) are believed to play opposing roles in host immunity against mycobacterial infection. IFN-gamma activates macrophages, while IL-10 downregulates the expression of T helper type 1 cytokines, MHC class II antigens and costimulatory molecules on macrophages. Associations of IFN-gamma -179 (G/T), +874 (A/T), +875 miscrosatellite CA repeats and +4766 (C/T), and IL-10 -1082 (A/G), -819 (C/T) and -592 (C/A) with tuberculosis (TB) were investigated in 385 HIV-negative patients and 451 controls in a Hong Kong Chinese population. The frequency of a low IFN-gamma-producing +874 A/A genotype was significantly over-represented in the patient group (P<0.001, OR=3.79, 95% CI=1.93-7.45). We identified 10 alleles in the IFN-gamma CA repeats and observed a significant difference in allele frequency distribution between patients and controls (P<0.001). By grouping alleles into 12 and non-12 CA repeats, the non-12/non-12 genotype yielded a similar significant result (P<0.001, OR=4.56, 95% CI=2.21-9.43) as observed in +874 A/A genotype. Weak associations of the IL-10 GCC/- genotype (P=0.04) and the low IFN-gamma-producing A/A genotype (P=0.06) with TB relapse/extrapulmonary cases were found. This study suggests the possible role of interferon gamma in TB susceptibility. PMID- 15815689 TI - Single nucleotide polymorphisms and haplotypes in the IL10 region associated with HCV clearance. AB - Hepatitis C virus (HCV) is an infectious blood-borne pathogen that usually persists as a chronic infection. However, approximately 15% of the time, patients can clear the virus, indicating that host differences could be critical in determining the course of HCV infection. The inflammatory response is crucial to resolving or failing to resolve an acute HCV infection. Some previous reports have implicated interleukin 10 (IL10) polymorphisms with successful anti-HCV therapy and natural viral clearance. We tested 54 single nucleotide polymorphisms (SNPs) in the IL10 region (+/-300 kb and 24 within the IL10 gene itself), which contains 13 genes including the IL10 immunomodulatory paralogs IL19, IL20, and IL24, for association with HCV clearance vs persistence. SNPs from two haplotype block regions, one at IL10 and the other from IL19/IL20, were associated with HCV clearance in African Americans (91 clearance cases and 183 chronically infected matched controls; P=0.05-0.002) while with expectation-maximization algorithm reconstructed haplotypes, these associations remained (P=0.05-0.002). However, no significant associations were detected in European Americans (108 clearance and 245 chronic). Our results indicate that variants of the immunomodulatory IL10 and IL19/IL20 genes may be involved in natural clearance of HCV in the African American population. PMID- 15815690 TI - Association of multiple sclerosis with ILT6 deficiency. AB - Multiple sclerosis (MS) is an autoimmune disorder of multifactorial etiology. Family studies have shown strong genetic contributions. Linkage analyses have revealed several regions harboring risk genes including chromosome region 19q13. ILT6 is one of the most interesting candidate genes, since ILTs are involved in the generation of immunological tolerance. There is an absence/presence of variability of the ILT6 gene comprising several exons, thus incapacitating the gene function. In the present study, we examined the association of ILT6 deletion with MS. Using PCR typing, deficiency of ILT6 was examined in 607 blood donors and in 751 Caucasian German, as well as 89 French MS patients. Homozygous ILT6 deficiencies were significantly more prevalent in MS patients (7.1%) than in blood donors (3.8%; P=0.009). ILT6 deficiency is associated with MS in the German population and hence a likely risk factor for autoimmune disorders. PMID- 15815691 TI - Novel IL-6 haplotypes and disease association. AB - Interleukin-6 (IL-6) is a pleiotropic cytokine crucial in both adaptive and innate immunity. Numerous genetic studies have shown association with variants of this gene in a multitude of diseases and phenotypes. Most tests of association have focused on a limited set of promoter polymorphisms, in particular, the 174G>C; however, there are many inconsistencies within and between these studies. We propose that there is a more complex regulatory haplotype extending further upstream of the previously characterised promoter region which will provide a more detailed view of the effect of variation on lL-6 regulation. We have exploited two additional single nucleotide polymorphisms (SNPs) in IL-6 that, when examined as a haplotype with existing markers, show an increased level of association with systemic onset juvenile arthritis in a family-based study. This suggests that the haplotype effect may be more functionally relevant to the disease. PMID- 15815692 TI - A new Fc receptor homolog, FREB2, found in germinal center B cells. AB - Fc receptor homologs are a recently identified family of proteins homologous to FcgammaRI, found on human and mouse B cells. One of these, FREB/FcRX/FCRL, was found to be unique since it lacks a transmembrane domain and is expressed intracellularly within germinal center B cells. We have identified in humans and mice a new Fc receptor homolog, FREB2, that blends conserved elements of the classical Fc gamma receptors with structural motifs previously thought to be unique to FREB1. This protein is comprised of three immunoglobulin-like domains with high homology to those in FcgammaRI, and a C-terminus containing a proline rich stalk region followed by a leucine-rich amphipathic alpha helix. Like FREB1, FREB2 is expressed as an intracellular protein. In murine splenocytes, RNA transcripts for each of the two proteins can be amplified from germinal center B cells. However, immunohistochemical analysis of human tonsils indicates that expression of FREB1 and FREB2 is mutually exclusive in non-neoplastic cells. Importantly, FREB2 expression within human tonsils appears to be limited to a small subset of nonproliferating germinal center B cells, suggesting that it may play a role in regulating clonal expansion or differentiation of B cells during the germinal center reaction. PMID- 15815693 TI - Detection of the CCR5-Delta32 HIV resistance gene in Bronze Age skeletons. AB - A mutant allele of the chemokine receptor CCR5 gene (CCR5-Delta32), which confers resistance to HIV-1 infection, is believed to have originated from a single mutation event in historic times, and rapidly expanded in Caucasian populations, owing to an unknown selective advantage. Among other candidates, the plague bacillus Yersinia pestis was implicated as a potential source of strong selective pressure on European populations during medieval times. Here, we report amplifications of the CCR5-Delta32 DNA sequence from up to 2900-year-old skeletal remains from different burial sites in central Germany and southern Italy. Furthermore, the allele frequency of CCR5-Delta32 in victims of the 14th century plague pandemic in Lubeck/northern Germany was not different from a historic control group. Our findings indicate that this mutation was prevalent already among prehistoric Europeans. The results also argue against the possibility of plague representing a major selective force that caused rapid increase in CCR5 Delta32 gene frequencies within these populations. PMID- 15815694 TI - Association between arterial stiffness and platelet activation. AB - Increased arterial stiffness is strongly associated with atherosclerosis, while platelet activation is an important trigger of thrombotic events in patients with atherosclerosis. However, little is known about the effect of arterial stiffness on platelet activation. We therefore investigated the association between arterial stiffness and platelet activation in 38 normal volunteers (20 men and 18 women) aged 23-77 years (mean = 49 +/- 15 years). Arterial stiffness was assessed by measuring brachial-ankle pulse wave velocity (ba-PWV) and heart-brachial PWV (hb-PWV). Flow cytometric analyses were performed to evaluate platelet activation by measuring surface expression of P-selectin and platelet-neutrophil complexes (PNC) before and after activation by ADP. We also calculated the difference between basal and stimulated states of P-selectin and PNC to assess platelet activation reserve. PWVs were significantly correlated with age and BP (r = 0.60 0.81). For platelet activation and activation reserve, correlations with age were less strong but remained significant (r = 0.36-0.61), with the exception of P selectin (not significant, NS), and correlations with SBP were similar (r = 0.35 0.53). A significant correlation was found between PWVs and platelet activation (r = 0.43-0.74). Multiple regression analysis demonstrated significant correlations between platelet activation and reserve and PWVs (coefficient = 2.17 6.59), when both age and BP were adjusted for simultaneously. In conclusion, platelet activation was associated with arterial stiffness, suggesting that arterial stiffness may play an important role in thrombotic events. PMID- 15815695 TI - Hypertension in patients with type-II diabetes: relation to urinary albumin excretion, endothelial function and inflammation. PMID- 15815696 TI - An interview with David Baltimore. [Interview by Joseph Glorioso and Jenny Jacoby]. PMID- 15815697 TI - Delivery into cells: lessons learned from plant and bacterial toxins. AB - A number of protein toxins of bacterial and plant origin have cytosolic targets, and knowledge about these toxins have provided us with essential information about mechanisms that can be used to gain access to the cytosol as well as detailed knowledge about endocytosis and intracellular sorting. Such toxins include those that have two moieties, one (the B-moiety) that binds to cell surface receptors and another (the A-moiety) with enzymatic activity that enters the cytosol, as well as molecules that only have the enzymatically active moiety and therefore are inefficient in cell entry. The toxins discussed in the present article include bacterial toxins such as Shiga toxin and diphtheria toxin, as well as plant toxins such as ricin and ribosome-inactivating proteins without a binding moiety, such as gelonin. Toxins with a binding moiety can be used as vectors to translocate epitopes, intact proteins, and even nucleotides into the cytosol. The toxins fall into two main groups when it comes to cytosolic entry. Some toxins enter from endosomes in response to low endosomal pH, whereas others, including Shiga toxin and ricin, are transported all the way to the Golgi apparatus and the ER before they are translocated to the cytosol. Plant proteins such as gelonin that are without a binding moiety are taken up only by fluid phase endocytosis, and normally they have a low toxicity. However, they can be used to test for disruption of endosomal membranes leading to cytosolic access of internalized molecules. Similarly to toxins with a binding moiety they are highly toxic when reaching the cytosol, thereby providing the investigator with an efficient tool to study endosomal disruption and induced transport to the cytosol. In conclusion, the protein toxins are useful tools to study transport and cytosolic translocation, and they can be used as vectors for transport to the interior of the cell. PMID- 15815698 TI - Gene therapy progress and prospects: recent progress in transgene and RNAi expression cassettes. AB - Plasmid expression cassette design must include a thoughtful analysis of potentially every nucleotide comprising a covalently closed circular or end protected linear DNA. This review will discuss recent studies in unraveling the mechanisms of postdelivery gene silencing, codon optimization and promoter identification. The recent discovery of potent RNA interference (RNAi) mechanisms for sequence-specific gene silencing has also invoked a great deal of interest in development of expression cassettes that can produce double-stranded RNA molecules for RNAi. Expression cassettes based on both RNA polymerase II and polymerase III transcription units that generate double-stranded RNA molecules for RNAi will also be discussed. PMID- 15815700 TI - Tf-lipoplex-mediated NGF gene transfer to the CNS: neuronal protection and recovery in an excitotoxic model of brain injury. AB - The development of efficient systems for in vivo gene transfer to the central nervous system (CNS) may provide a useful therapeutic strategy for the alleviation of several neurological disorders. In this study, we evaluated the feasibility of nonviral gene therapy to the CNS mediated by cationic liposomes. We present evidence of the successful delivery and expression of both a reporter and a therapeutic gene in the rodent brain, as evaluated by immunohistochemical assays. Our results indicate that transferrin-associated cationic liposome/DNA complexes (Tf-lipoplexes) allow a significant enhancement of transfection activity as compared to plain complexes, and that 8/1 (+/-) Tf-lipoplexes constitute the best formulation to mediate in vivo gene transfer. We demonstrated that Tf-lipoplex-mediated nerve growth factor transgene expression attenuates the morphological damages of the kainic acid-induced lesion as assessed by 2,3,5 triphenyltetrazolium chloride (TTC) vital staining. These findings suggest the usefulness of these lipid-based vectors in mediating the delivery of therapeutic genes to the CNS. PMID- 15815701 TI - Enhanced repair of articular cartilage defects in vivo by transplanted chondrocytes overexpressing insulin-like growth factor I (IGF-I). AB - Traumatic articular cartilage lesions have a limited capacity to heal. We tested the hypothesis that overexpression of a human insulin-like growth factor I (IGF I) cDNA by transplanted articular chondrocytes enhances the repair of full thickness (osteochondral) cartilage defects in vivo. Lapine articular chondrocytes were transfected with expression plasmid vectors containing the cDNA for the Escherichia coli lacZ gene or the human IGF-I gene and were encapsulated in alginate. The expression patterns of the transgenes in these implants were monitored in vitro for 36 days. Transfected allogeneic chondrocytes in alginate were transplanted into osteochondral defects in the trochlear groove of rabbits. At three and 14 weeks, the quality of articular cartilage repair was evaluated qualitatively and quantitatively. In vitro, IGF-I secretion by implants constructed from IGF-I-transfected chondrocytes and alginate was 123.2+/-22.3 ng/10(7) cells/24 h at day 4 post transfection and remained elevated at day 36, the longest time point evaluated. In vivo, transplantation of IGF-I implants improved articular cartilage repair and accelerated the formation of the subchondral bone at both time points compared to lacZ implants. The data indicate that allogeneic chondrocytes, transfected by a nonviral method and cultured in alginate, are able to secrete biologically relevant amounts of IGF-I over a prolonged period of time in vitro. The data further demonstrate that implantation of these composites into deep articular cartilage defects is sufficient to augment cartilage defect repair in vivo. These results suggest that therapeutic growth factor gene delivery using encapsulated and transplanted genetically modified chondrocytes may be applicable to sites of focal articular cartilage damage. PMID- 15815702 TI - Correction of a mineralization defect by overexpression of a wild-type cDNA for COL1A1 in marrow stromal cells (MSCs) from a patient with osteogenesis imperfecta: a strategy for rescuing mutations that produce dominant-negative protein defects. AB - Gene therapy for dominant-negative disorders presents a more difficult challenge than gene therapy for recessive disorders, since even partial replacement of a protein for a recessive disorder can reverse symptoms. Osteogenesis imperfecta (OI) has frequently served as a model disorder for dominant-negative defects of structural proteins. The disease is caused by mutations in type I collagen (COL1A1), the major structural component of bone, skin and other connective tissues. The severity of the phenotype is largely dependent on the ratio of normal to mutant type I procollagen synthesized by cells. Recently, attempts have been made to develop strategies for cell and gene therapies using the adult stem cells from bone marrow referred to as mesenchymal stem cells or marrow stromal cells (MSCs). In this study, we used MSCs from a patient with type III OI who was heterozygous for an IVS 41A+4C mutation in COL1A1. A hybrid genomic / cDNA construct of COL1A1 was transfected into the MSCs and the transfectants were expanded over a 200-fold. Transfected MSCs showed increased expression of the wild-type mRNA and protein. In vitro assays demonstrated that the transfected cells more efficiently differentiated into mineralizing cells. The results indicated that it is possible to overexpress COL1A1 cDNA in OI MSCs and thereby to correct partially the dominant-negative protein defect. PMID- 15815703 TI - Quantitative comparison of polyethylenimine formulations and adenoviral vectors in terms of intracellular gene delivery processes. AB - An objective of designing molecular vehicles exhibiting virus-like transgene delivery capabilities but with low toxicity and immunogenicity continues to drive synthetic vector development. As no single step within the gene delivery pathway represents the critical limiting barrier for all vector types under all circumstances, improvements in synthetic vehicle design may be aided by quantitative analysis of the contributions of each step to the overall delivery process. To our knowledge, however, synthetic and viral gene delivery methods have not yet been explicitly compared in terms of these delivery pathway steps in a quantitative manner. As a first address of this challenge, we compare here quantitative parameters characterizing intracellular gene delivery steps for an E1/E3-deleted adenoviral vector and three polyethylenimine (PEI)-based vector formulations, as well as the liposomal transfection reagent Lipofectamine and naked DNA; the cargo is a plasmid encoding the beta-galactosidase gene under a CMV promoter, and the cell host is the C3A human hepatocellular carcinoma line. The parameters were determined by applying a previously validated mathematical model to transient time-course measurements of plasmid uptake and trafficking (from whole-cell and isolated nuclei lysates, by real-time quantitative PCR), and gene expression levels, enabling discovery of those for which the adenoviral vector manifested superiority. Parameter-sensitivity analysis permitted identification of processes most critically rate-limiting for each vector. We find that the adenoviral vector advantage in delivery appears to reside partially in its import to the nuclear compartment, but that its vast superiority in transgene expression arises predominantly in our situation from postdelivery events: on the basis of per-nuclear plasmid, expression efficiency from adenovirus is superior by orders of magnitude over the PEI vectors. We find that a chemical modification of a PEI-based vector, which substantially improves its performance, appears to do so by enhancing certain trafficking rate parameters, such as binding and uptake, endosomal escape, and binding to nuclear import machinery, but leaves endosomal escape as a barrier over which transgene delivery could be most sensitively increased further for this polymer. PMID- 15815704 TI - Primary adenovirus-specific cytotoxic T lymphocyte response occurs after viral clearance and liver enzyme elevation. AB - The virus-specific cytotoxic T lymphocyte (CTL) response is a major obstacle to effective delivery of adenovirus gene therapy. However, its relative role in viral clearance, transgene elimination and hepatotoxicity remains unclear. In this paper, we present an analysis of viral clearance and liver toxicity in relation to the induction of the virus-specific CD8 T-cell response revealed by an MHC class I tetramer. A surprisingly high number of tetramer+ CD8 T cells were found in the liver and lung and reached peak values at days 8 and 10, respectively, post-infection. Nearly 100% of these tetramer+ CD8 T cells expressed high levels of granzyme B and IFNgamma. Remarkably, liver viral load and liver enzyme elevation peaked early, at days 2 and 4, respectively, post infection, before the specific CTL response was detectable. After generation of CTLs, there was only minimal liver damage or further decrease in virus titer. These results indicated that the primary peak response of tetramer+ CTLs does not correlate with the elimination of adenovirus or liver cytotoxic response. PMID- 15815705 TI - Engineered E. coli delivers therapeutic genes to the colonic mucosa. AB - Taking advantage of the proximity of bowel mucosa to luminal bacteria, we have attempted to deliver a therapeutic gene to the colonic mucosa by oral administration of an invasive and non-pathogenic Escherichia coli. E. coli diamenopimelate (dap) auxotroph, harboring plasmid pGB2Omegainv-hly, express the inv gene from Yersinia pseudotubercolosis that confers the ability to invade nonprofessional phagocytic cells and the hly gene from Listeria monocytogenes that allows expression of lystreriolysin O, a perforin cytolysin able to perfore phagosomal membranes. This bacterial vector invades and transfers functional DNA to epithelial cells in vitro. We have shown that this strain carrying a therapeutic gene (pC1OmegaTGF-beta1) can significantly reduce the severity of experimental colitis in mice. However, as a consequence of mucosal barrier disruption during colitis, vector-specific mRNA transcripts could be recovered from the colon and also from extra-colonic tissues. We therefore replaced the constitutive CMV promoter in pC1OmegaTGF-beta1 by the inflammation-inducible interleukin-8 promoter generating plasmid pC1OmegaTGF-beta1IND. Plasmid-specific TGF-beta1 mRNA transcripts were detectable in mouse CMT-93 epithelial cells incubated with E. coli BM2710/pGB2Omegainv-hly carrying pC1OmegaTGF-beta1IND following exposure to inflammatory cytokines. Furthermore, the transcripts were detectable only within inflamed tissues and the therapeutic effects were comparable to those in animals treated with E. coli BM2710/pGB2Omegainv hly+pC1OmegaTGF-beta1. In summary, engineered enteric bacteria can efficiently deliver in vivo therapeutic genes to the intact intestinal mucosa and regulation expression of the therapeutic gene by an inflammation-inducible promoter prevents its dissemination during colitis. PMID- 15815707 TI - The hemodynamic effects of neonatal patent ductus arteriosus shunting on superior mesenteric artery blood flow. AB - OBJECTIVE: To determine if the ratio of the pulsatility index (PI) of the left pulmonary artery to the PI of the descending aorta, the Rp/Rs index, correlates with the degree of ductal steal from the intestine in neonates with a patent ductus arteriosus (PDA). STUDY DESIGN: Echocardiograms and Doppler studies of the superior mesenteric artery (SMA) were performed in 41 neonates less than 35 weeks gestational age with a hemodynamically significant PDA (hsPDA). RESULTS: There was a significant negative correlation between the Rp/Rs index and the SMA PI after controlling for ductal size (r=-0.476, p<0.008). CONCLUSIONS: The Rp/Rs index can be used as an indicator of ductal steal on intestinal blood flow. The Rp/Rs index may be a useful adjunct to existing and new techniques for improving early assessment and treatment of hsPDA, and for evaluating the effects of hsPDA on systemic organs. PMID- 15815706 TI - HIV-1-derived lentiviral vectors and fetal route of administration on transgene biodistribution and expression in rhesus monkeys. AB - The gene transfer efficiency of lentiviral vectors pseudotyped with vesicular stomatitis virus-glycoprotein (VSV-G) driven by the MND or CMV promoters and expressing the enhanced green fluorescent protein (EGFP) was investigated in fetal rhesus monkeys (Macaca mulatta) (N=21). Fetuses (50+/-10 days gestation; term 165+/-10 days) were injected under ultrasound guidance using an intraperitoneal (i.p.) or intrahepatic (i.h.) approach with a range of 1 x 10(7) 2 x 10(8) infectious particles/fetus. Analysis of transgene biodistribution and expression was performed in multiple tissues at 3-7 months postgene delivery using quantitative techniques. Overall, results indicated the following: (1) i.p. gene transfer at 40 days gestation resulted in a more diffuse distribution of the vector compared to administration at 60 days gestation; (2) vector biodistribution was similar after administration by the i.p. or i.h. routes; and (3) gene expression analysis in transduced tissues showed the presence of mRNA transcripts that correlated with the level of gene transfer. These studies suggest that fetal gene transfer using the i.p. and i.h. routes results in prolonged transduction and expression of the transgene in multiple tissues. PMID- 15815708 TI - Amplitude-integrated EEG in preterm infants: maturation of background pattern and amplitude voltage with postmenstrual age and gestational age. AB - OBJECTIVE: Amplitude-integrated electroencephalogram (aEEG) is a single channel EEG recorded from two parietal electrodes. The objective of this study was to test the hypothesis that aEEG maturation follows postmenstrual age (PMA) irrespective of gestational age (GA). METHODS: We recruited inborn infants with a GA <33 weeks and without evidence of neurologic anomaly. Serial aEEG recordings were assessed for: presence of continuous activity and mature sleep-wake cycling (SWC); low base voltage (V), that is, the lowest amplitude margin; high base V, that is, the most common amplitude margin; upper high V, that is, upper margin during highest activity; and span, that is, the difference between upper high and simultaneous high base V. Statistical analysis included logistic regression and repeated measures analysis of variance. RESULTS: We obtained 119 aEEG recordings in 31 preterm infants (GA 25 to 32 weeks; birth weight 600 to 1704 g, PMA 25 to 35 weeks). The frequency of mature SWC increased with PMA independent of GA, while the frequency of continuity increased with PMA and was higher in extremely preterm infants after correcting for PMA. Low base and high base V increased with PMA, while span and upper high V significantly decreased with PMA. In addition, high base V was higher in extremely preterm infants after correcting for PMA. CONCLUSIONS: In preterm infants aEEG matures predominantly with PMA. Our data suggest that some aspects of aEEG maturation are enhanced, rather than inhibited by extremely preterm birth. These data suggest that aEEG in preterm infants may need to be analyzed by comparing results with standards of similar PMA and GA. PMID- 15815709 TI - Evolution and development: Wnts in deep time. PMID- 15815710 TI - Immune function responds to selection for cuticular colour in Tenebrio molitor. AB - Cuticular colour in the mealworm beetle (Tenebrio molitor) is a quantitative trait, varying from tan to black. Population level variation in cuticular colour has been linked to pathogen resistance in this species and in several other insects: darker individuals are more resistant to pathogens. Given that cuticular colour has a heritable component, we have taken an experimental evolution approach: we selected 10 lines for black and 10 lines for tan adult cuticular phenotypes over at least six generations and measured the correlated responses to selection in a range of immune effector systems. Our results show that two immune parameters related to resistance (haemocyte density and pre-immune challenge activity of phenoloxidase (PO)) were significantly higher in selection lines of black beetles compared to tan lines. This may help to explain increased resistance to pathogens in darker individuals. Cuticular colour is dependent upon melanin production, which requires the enzyme PO that is present in its inactive form inside haemocytes. Thus, the observed correlated response to selection upon cuticular colour and immune variables probably results from these traits' shared dependence on melanin production. PMID- 15815711 TI - Insulators are fundamental components of the eukaryotic genomes. AB - The properties of cis-regulatory elements able to influence gene transcription over large distances have led to the hypothesis that elements called insulators should exist to limit the action of enhancers and silencers. During the last decades, insulators have been identified in many eukaryotes from yeast to human. Insulators possess two main properties: (i) they can block enhancer-promoter communication ('enhancer blocker activity'), and (ii) they can prevent the spread of repressive chromatin ('barrier activity'). This review focuses on recent studies designed to elucidate the molecular mechanisms of the insulator function, and gives an overview of the critical role of insulators in nuclear organization and functional identity of chromatin. PMID- 15815712 TI - Genetic evidence for a family-based Scandinavian settlement of Shetland and Orkney during the Viking periods. AB - The Viking age witnessed the expansion of Scandinavian invaders across much of northwestern Europe. While Scandinavian settlements had an enduring cultural impact on North Atlantic populations, the nature and extent of their genetic legacy in places such as Shetland and Orkney is not clear. In order to explore this question further, we have made an extensive survey of both Y-chromosomal and mitochondrial DNA (mtDNA) variation in the North Atlantic region. Our findings indicate an overall Scandinavian ancestry of approximately 44% for Shetland and approximately 30% for Orkney, with approximately equal contributions from Scandinavian male and female subjects in both cases. This contrasts with the situation for the Western Isles, where the overall Scandinavian ancestry is less ( approximately 15%) and where there is a disproportionately high contribution from Scandinavian males. In line with previous studies, we find that Iceland exhibits both the greatest overall amount of Scandinavian ancestry (55%) and the greatest discrepancy between Scandinavian male and female components. Our results suggest that while areas close to Scandinavia, such as Orkney and Shetland, may have been settled primarily by Scandinavian family groups, lone Scandinavian males, who later established families with female subjects from the British Isles, may have been prominent in areas more distant from their homeland. PMID- 15815713 TI - Spatial genetic structure of two sympatric neotropical palms with contrasting life histories. AB - The spatial genetic structure within sympatric populations of two neotropical dioecious palm species with contrasting life histories was characterized to evaluate the influence of life history traits on the extent of genetic isolation by distance. Chamaedorea tepejilote is a common wind-pollinated arboreal understory palm. Chamaedorea elatior is an uncommon climbing subcanopy palm with entomophilous pollination syndrome. A total of 59 allozyme alleles for C. tepejilote and 53 alleles for C. elatior was analyzed using both unweighted (Iu) and weighted (Iw) Moran's I spatial autocorrelation statistics. The spatial genetic structure detected within these populations is consistent with those reported for highly dispersed plants. A significance test for differences between mean Moran's I-coefficients revealed less spatial genetic structure within the C. tepejilote population than that in the C. elatior population. Adjacent individuals of C. elatior exhibited significant spatial genetic autocorrelation (Iu=0.039, Iw=0.034), indicating a Wright's neighborhood size of about 100 individuals. For C. tepejilote, nonrandom genetic distribution among nearest neighbors was detected, even from small spatial autocorrelation values (Iu=0.008, Iw=0.009), consistent with a neighborhood size of about 300 individuals. For both species, seed dispersal, mortality among life cycle stages, overlapping generations, and contrasting traits of mating and reproduction influence the standing spatial genetic structure within populations. PMID- 15815714 TI - Thrombocytopenia after gemtuzumab is reversible by intravenous immunoglobulin. PMID- 15815715 TI - CBFbeta-SMMHC slows proliferation of primary murine and human myeloid progenitors. AB - CBFbeta-SMMHC is expressed in 8% of acute myeloid leukemias and inhibits AML1/RUNX1. In this study, murine marrow or human CD34(+) cells were transduced with retroviral or lentiviral vectors expressing CBFbeta-SMMHC or two mutant variants. CBFbeta-SMMHC reduced murine or human myeloid cell proliferation three- to four-fold in liquid culture relative to empty vector-transduced cells, during a period when vector-transduced cells accumulated five-fold and human cells 20 fold. CBFbeta-SMMHC decreased the formation of myeloid, but not erythroid, colonies two- to four-fold, and myeloid colonies expressing CBFbeta-SMMHC were markedly reduced in size. However, CBFbeta-SMMHC did not slow differentiation to granulocytes or monocytes. Neither CBFbeta-SMMHC(Delta2-11), which does not bind AML1, nor CBFbeta-SMMHC(DeltaACD), which does not multimerize or efficiently bind corepressors, slowed proliferation or reduced myeloid colonies. CBFbeta-SMMHC increased the G1/S ratio 1.4-fold. AML1 had an effect opposite to CBFbeta-SMMHC, stimulating proliferation of murine myeloid progenitors 2.0-fold in liquid culture. Thus, CBFbeta-SMMHC directly inhibits the proliferation of normal myeloid progenitors via inhibition of AML1 and dependent upon the integrity of its assembly competence domain. These findings support the development of therapeutics that target the ability of CBFbeta-SMMHC to interact with AML1 or to multimerize via its assembly competence domain. PMID- 15815716 TI - Improved therapy of non-Hodgkin's lymphoma xenografts using radionuclides pretargeted with a new anti-CD20 bispecific antibody. AB - A comparison of the therapeutic efficacy of a new bispecific monoclonal antibody (bsMAb)-pretargeting system vs the conventional direct targeting modality was undertaken. A bsMAb was made by coupling the Fab' of a humanized anti-CD20 antibody to the Fab' of a murine antibody directed against the peptide histamine succinyl-glycine (HSG). The tumor targeting of the bsMAb was separated from the subsequent delivery of the radionuclide-bearing HSG peptide conjugated with (111)In or (90)Y. Nude mice bearing s.c. Ramos human B-cell lymphomas were injected with the bsMAb and then, 48 h later, (111)In/(90)Y-HSG peptide was given. At 3 h postinjection, tumor/blood ratios for pretargeted (111)In-HSG peptide were similar to that observed with the directly conjugated (111)In-anti CD20 IgG at its highest level on day 7, but by day 1, tumor/blood ratios were about 10-fold higher than the IgG. Tumors progressed rapidly in animals given 800 microCi of (90)Y-HSG peptide alone, whereas 5/10 animals in the group pretargeted by the anti-CD20 bsMAb were tumor-free 18 weeks later. The antitumor response in animals administered the pretargeted (90)Y-HSG peptide was also significantly superior to treatment with the directly radiolabeled (90)Y-anti-CD20 IgG, whether given as a single injection (P<0.007) or as a divided dose (P=0.016). This bsMAb pretargeting procedure significantly improves the therapeutic response of targeted radionuclides in non-Hodgkin's lymphoma, warranting further development of this method of radioimmunotherapy. PMID- 15815717 TI - Apoptosis-resistant phenotype of classical Hodgkin's lymphoma is not mediated by somatic mutations within genes encoding members of the death-inducing signaling complex (DISC). PMID- 15815718 TI - New insights into MLL gene rearranged acute leukemias using gene expression profiling: shared pathways, lineage commitment, and partner genes. AB - Rearrangements of the MLL gene occur in both acute lymphoblastic and acute myeloid leukemias (ALL, AML). This study addressed the global gene expression pattern of these two leukemia subtypes with respect to common deregulated pathways and lineage-associated differences. We analyzed 73 t(11q23)/MLL leukemias in comparison to 290 other acute leukemias and demonstrate that 11q23 leukemias combined are characterized by a common specific gene expression signature. Additionally, in unsupervised and supervised data analysis algorithms, ALL and AML cases with t(11q23) segregate according to the lineage they are derived from, that is, myeloid or lymphoid, respectively. This segregation can be explained by a highly differing transcriptional program. Through the use of novel biological network analyses, essential regulators of early B cell development, PAX5 and EBF, were shown to be associated with a clear B-lineage commitment in lymphoblastic t(11q23)/MLL leukemias. Also, the influence of the different MLL translocation partners on the transcriptional program was directly assessed. Interestingly, gene expression profiling did not reveal a clear distinct pattern associated with one of the analyzed partner genes. Taken together, the identified molecular expression pattern of MLL fusion gene samples and biological networks revealed new insights into the aberrant transcriptional program in 11q23/MLL leukemias. PMID- 15815719 TI - t(11;14) does not predict long-term survival in myeloma. PMID- 15815720 TI - Identification of an HLA-A1 restricted CTL epitope from Mcl-1. PMID- 15815722 TI - Homozygous deletion of SOCS1 in primary mediastinal B-cell lymphoma detected by CGH to BAC microarrays. PMID- 15815724 TI - Imatinib induces apoptosis in CLL lymphocytes with high expression of Par-4. PMID- 15815725 TI - Low-grade follicular lymphoma with t(14;18) presents a homogeneous disease entity otherwise the rest comprises minor groups of heterogeneous disease entities with Bcl2 amplification, Bcl6 translocation or other gene aberrances. AB - Follicular lymphomas (FL) are morphologically classified into grades 1, 2, 3a and 3b by the World Health Organization. Bcl2, Bcl6 and CD10 are phenotypic markers of FL while the Bcl2 t(14;18) and Bcl6 t(3q27) gene translocations are common genetic changes. However, to date, there has been no integrated analysis based on phenotype, grade and genotype from large numbers of FL cases. We graded 261 cases of FL and determined their phenotypes and gene alterations. According to the antigen markers and gene alterations of 147 cases, we classified FL into typical and the others types. The typical group, which includes 69% cases of FL, is characterized by low histological grade (grade 1, 2), coexpression of BCL2 and CD10 and Bcl2 gene translocation. The rest comprises a small part of low-grade FL without Bcl2 gene translocation and high-grade (grade 3a, 3b) FL. These FLs include some heterogeneous disease entities. They are characterized by high histological grade (87%), no definite expression of BCL2 or CD10 and several kinds of gene aberrances including Bcl2 translocation, Bcl6 translocation, Bcl2 amplification or other unknown gene abnormality. Our findings indicate that typical FL presents a homogeneous disease entity whereas the rest comprises heterogeneous diseases entities. PMID- 15815726 TI - Identification of Ki23819, a highly potent inhibitor of kinase activity of mutant FLT3 receptor tyrosine kinase. AB - Constitutively active internal tandem duplication (ITD) in the juxtamembrane domain of Fms-like tyrosine kinase 3 (FLT3), a type III receptor tyrosine kinase, is the most common molecular defect associated with acute myeloid leukemia. Its presence confers a poor outcome in patients with acute myeloid leukemia who receive conventional chemotherapy. FLT3-ITD has therefore been considered to be an attractive molecular target for a novel therapeutic modality. We describe here the identification and characterization of Ki23819 as a novel FLT3 inhibitor. Ki23819 suppressed proliferation and induced apoptosis of FLT3-ITD-expressing human leukemia cell lines. The growth-inhibitory effect of Ki23819 on MV4-11 cells was superior to that of SU11248, another FLT3 inhibitor (IC(50)<1 vs 3-10 nM). Ki23819 inhibited the autophosphorylation of FLT3-ITD more efficiently than that of wild-type FLT3. FLT3-ITD-dependent activation of the downstream signaling proteins ERK and STAT5 was also inhibited within similar concentration ranges. Thus, Ki23819 is a potent in vitro inhibitor of FLT3. PMID- 15815727 TI - Identification of genes potentially involved in disease transformation of CML. AB - In patients with chronic myeloid leukemia (CML) who do not reach a (near) complete cytogenetic response, the disease progresses over several years from an indolent, chronic phase into a rapidly fatal blast crisis. Events that are responsible for this transformation process are largely unknown. To identify changes in gene expression that occurred during the course of the disease, we performed cDNA subtraction on sequentially stored peripheral blood mononuclear cell pellets, collected throughout the course of disease of a single CML patient. In total, 32 differentially expressed sequences were identified, of which 27 corresponded to known genes. On quantitative PCR, eight of these genes, YWHAZ, GAS2, IL8, IL6, PBEF1, CCL4, SAT and MMRN, showed comparable differential expression in additional CML patient samples. This set of genes can be considered as a starting point for further research on causes of disease transformation in CML and may lead to new targets in the treatment of resistant CML. PMID- 15815729 TI - [What are Norwegian researchers working with?]. PMID- 15815728 TI - Nonproliferating CML CD34+ progenitors are resistant to apoptosis induced by a wide range of proapoptotic stimuli. AB - Imatinib mesylate, a Bcr-Abl kinase inhibitor, has been very successful in the treatment of chronic myelogenous leukemia (CML). However, the majority of patients achieving cytogenetic remissions with imatinib treatment have molecular evidence of persistent disease, and residual BCR/ABL(+) progenitors can be detected. There is a need to develop new approaches that enhance elimination of malignant progenitors in imatinib-treated patients. Here we show that CML CD34(+) progenitors are sensitive to several apoptosis-inducing stimuli including the chemotherapeutic agents Ara-C and VP-16, radiation, arsenic trioxide, ceramide, growth factor withdrawal, and the death receptor activators TNFalpha and TRAIL. Bcr-Abl kinase inhibition by imatinib did not enhance sensitivity of CML progenitors to Ara-C, VP-16, ceramide, radiation or TRAIL-induced apoptosis but did enhance arsenic and TNFalpha-induced apoptosis. We further demonstrate that apoptosis was restricted to dividing cells, whereas nonproliferating BCR/ABL(+) CD34(+) cells were resistant to apoptosis induced by imatinib, Ara-C or arsenic, either alone or in combination. Resistance of quiescent CML progenitors to imatinib-induced apoptosis could contribute to persistence of residual malignant progenitors in imatinib-treated patients. Combination treatment with Ara-C or arsenic may not enhance targeting of nonproliferating CML progenitors. The assay described here may be useful for identifying agents targeting quiescent CML progenitors. PMID- 15815730 TI - [Epicrisis--an invoice or a medical record?]. PMID- 15815731 TI - [To cure chronic bed sores]. PMID- 15815732 TI - [Treatment of patients with gastrointestinal stromal tumour with imatinib mesylate]. AB - BACKGROUND: Gastrointestinal stromal tumour (GIST) is the most frequent mesenchymal tumour type of the digestive tract. Between 30 and 40% of patients have high-risk, malignant GIST with poor prognosis after surgery. Imatinib mesylate is a recently introduced KIT tyrosine kinase inhibitor with effect on metastatic GIST. We report our experience with imatinib mesylate in the treatment of GIST. MATERIAL AND METHODS: Nine patients diagnosed with GIST have received imatinib mesylate since August 2001. Eight patients had metastatic disease, one patient received adjuvant treatment. The patients were evaluated according to standard protocols for clinical performance, effect of treatment, and adverse effects. Tumour tissue was analysed for mutational status in KIT and PDGFRA. RESULTS: All patients with metastatic disease had palliative benefit; three had partial response and the remaining stable disease. The single patient receiving adjuvant treatment had no sign of recurrence. Side effects were mainly mild diarrhoea, nausea and vomiting. Seven patients had mutations in KIT exon 11, one in KIT exon 9, and one in PDGFRA exon 12. INTERPRETATION: The results demonstrate that imatinib mesylate is an effective drug that can stabilise and reduce disease in patients with advanced GIST. PMID- 15815733 TI - [COX-2 inhibitors--one step forward and two steps back]. AB - BACKGROUND: Selective cyclooxygenase-2 (COX-2) inhibitors are widely used. They have no advantages in terms of efficacy, and it is not documented that they cause fewer adverse effects than conventional NSAIDs. MATERIAL AND METHODS: The adverse effects of rofecoxib, celecoxib and other NSAIDs are reviewed. Relevant literature was identified on Medline and in the reference lists in key articles. RESULTS: Rofecoxib and the novel COX-2 inhibitors etoricoxib and valdecoxib have a higher degree of COX-2 selectivity than traditional NSAIDs. Celecoxib is less COX-2 selective and appears to be similar to diclofenac. Rofecoxib induces thromboembolic adverse effects more frequently than conventional NSAIDs. INTERPRETATION: The cardiovascular problems conferred by rofecoxib are probably a class effect and thus inducible by other selective COX-2 inhibitors. Pending comprehensive safety data, caution is warranted regarding the use of these drugs. PMID- 15815734 TI - [Should older men be treated with testosterone?]. AB - BACKGROUND: Male hypogonadism is a clinical situation characterised by a low serum testosterone level in combination with a diversity of symptoms and signs such as reduced libido and vitality, decreased muscle mass, increased fat mass and depression. Similar symptoms in combination with subnormal testosterone levels are seen in some elderly men. Several publications have suggested that testosterone treatment in hypogonadal men may have beneficial effects, but it is still uncertain whether testosterone substitution in the aging man is indicated. Despite this uncertainty the sale of testosterone have increased enormously over the last few years, hence it seems important to discuss what we now know about such treatment. MATERIAL AND METHODS: Review of recent literature studying testosterone substitution in older men. RESULTS AND INTERPRETATION: The result from placebo-controlled studies of testosterone substitution in elderly men differ substantially, but it seems to improve, among other things, bone mineral density, body composition, perception of physical strength, and maybe libido. In the short term there have been few problems or complications with testosterone treatment, but effects on the cardiovascular system and the prostate over the long term remain uncertain. Before any general recommendation could be given, big prospective studies have to be performed. Treatment should, however, be considered in men with testosterone in the hypogonadal range accompanied by clinical symptoms. Treatment needs to be individualized and should preferably be initiated by specialists in andrology, endocrinology or urology. PMID- 15815735 TI - [Surgery in patients with bleeding disorders--expensive treatment for a small group of patients]. AB - BACKGROUND: During surgical procedures, patients with bleeding disorders have a major risk of bleeding complications. To reduce the risk of bleeding it is necessary to provide pre-, peri- and postoperative antihaemorrhagic therapy. In less severe bleeding disorders, pharmacologic treatment may be sufficient, but in patients with severe bleeding disorders there is always a need for clotting factor concentrates. MATERIALS AND METHODS: This study includes all patients with bleeding disorders admitted to Rikshospitalet University Hospital between 1997 and 2003 for surgical procedures during which therapy with clotting factor concentrates was mandatory. RESULTS AND INTERPRETATION: Over the study period, 135 patients underwent a total of 255 different surgical procedures. In 47% of the patients there was a causal relationship between the need for surgery and the bleeding disorder. Our results show that patients with severe bleeding disorders, including patients with inhibitors, can be treated safely provided that the patients receive adequate treatment with clotting factor concentrates. However, substitution therapy with clotting factor concentrates is very expensive. Cost related to substitution therapy is the main determinant of how many patients with severe bleeding disorders can undergo elective surgery in our hospital each year. PMID- 15815736 TI - [Basic pharmacokinetics--absorption]. PMID- 15815737 TI - [Venous leg ulcers]. AB - Patients with venous leg ulcers usually have extensive symptoms both related to their venous insufficiency and to the wound itself, often combined with a reduced quality of life. Prevalence of venous leg ulcers varies from 0.1 to 1.0%. Treatment costs are high and may amount to 1.5% of a nation's total spending on health care. Venous hypertension is the common denominator for all patients with venous leg ulcers. Isolated superficial as well as deep or combined venous insufficiency with or without insufficient perforators may cause ulceration. In the microcirculation, inflammation is involved, but the exact mechanisms behind the ulcer formation remain unresolved. During the examination, a presence of superficial venous insufficiency accessible for superficial resection must be established. In addition to a clinical examination, venous pressure measurements/plethysmography and colour duplex scanning is recommended in order to locate and evaluate the significance of the venous insufficiency. The key element in the treatment of venous ulcers is to reduce oedema and venous hypertension by adequate compression and elevation. If primary superficial venous insufficiency is established, venous resection is recommended. This may improve healing and reduce recurrences. In selected patients, deep venous reconstruction is an alternative approach. PMID- 15815738 TI - [Arterial ischemic ulcers]. AB - Arterial ischaemic ulcers develop because of inadequate perfusion leading to local ischaemia in the skin and underlying tissue. The most common cause is peripheral arterial disease, giving rise to symptoms like intermittent claudication, rest pain and gangrene, in addition to local ulceration. Diabetes mellitus increases the risk of ulcer formation; admittedly mainly neuropathic ulcers with a low component of peripheral arterial disease. Yet a combination of neuropathy and ischaemia is common ("neuro-ischaemic ulcer"). A thorough patient history and clinical examination can help discriminate arterial ulcers from venous, pressure, traumatic and vasculitis ulcers. Reduction of ankle systolic pressure and calculated ankle/brachial index, sometimes additional other non invasive laboratory tests, confirm peripheral arterial disease. The primary treatment of arterial ischaemic ulcer is to increase blood supply to the affected area, primarily by endovascular treatment or open arterial reconstruction. Endovascular treatment (balloon angioplasty) is the method of choice because of graft infection risk in patients with open ulcers. Most arterial ischaemic ulcers will progress to healing if the blood supply is reestablished. PMID- 15815739 TI - [Diabetic foot ulcers]. AB - Diabetic foot ulcers result from two specific complications: neuropathy and arteriosclerosis. Prompt and appropriate treatment of these often complicated wounds can help reduce complications that may lead to amputation. The most effective way to prevent these complications is to refer patients to interdisciplinary diabetic foot teams. The most important aspects of treatment are off-loading, treatment of infections, establishment of sufficient circulation, and local wound therapy. A specific examination is the basis for risk assessment. Prophylaxis and appropriate management of patients at risk of developing foot ulcers would contribute to reduce the number of amputations among diabetic patients. PMID- 15815740 TI - [Medical records, DRG and intensive care patients]. AB - BACKGROUND: In order to control the quality of the medical report after a hospital stay with regards to the stay in the intensive care unit (ICU), and to cheque for correct DRG grouping, this study of 428 patients treated in our ICU in 2003 was conducted. MATERIAL AND METHODS: All ICU patients from 2003 were found in our database, which includes specific ICD-10 diagnosis and specific ICU procedures. The medical record summarising the hospital stay (epicrisis) was retrieved for each patient from the hospital's electronic patient files and controlled for correct information regarding the ICU stay. DRG groups for each patient were retrieved from the hospital's administrative database. All stays were re-coded, with all information about the ICU stay was also included. The new DRG codes were compared with the old ones, and the difference in DRG points computed. RESULTS: The description of the stay in the ICU was missing or very insufficient in 46% of the records. In the DRG control we found that an additional 347.37 DRG points (18.4% of the original sum of all DRG points) were missing, corresponding to a loss to the hospital of 6.2 million NOK. In addition we discovered missing codes for tracheostomy corresponding to 2.8 million NOK, giving a total loss of 9 million NOK. CONCLUSION: This study confirms that an adequate description of the stay in the ICU is insufficient in a large number of medical records. This also leads to incorrect DRG grouping of many patients and significant financial losses to the hospital. PMID- 15815743 TI - [World health]. PMID- 15815741 TI - [Care for children with tracheostomy]. AB - BACKGROUND: Modern technology has improved the quality of life for children with tracheostomy and home ventilators. MATERIAL AND METHODS: A "home ventilator team" at St. Olav University Hospital was responsible for 18 children with tracheostomy over a period of 2.5 years. The team serves as a link between different health care providers. A new educational programme for health care providers and parents is described. RESULTS AND INTERPRETATION: Children with tracheostomy constitute a small, but professionally challenging group with a variety of health-related problems. In Norway it is not quite clear who is responsible for the care of these patients. PMID- 15815744 TI - [How to keep one's own independence]. PMID- 15815745 TI - [Should acetylsalicylic acid be withdrawn prior to surgery?]. PMID- 15815746 TI - [Should patients be allowed to make notes in own medical records?]. PMID- 15815749 TI - [Thiazides one more time]. PMID- 15815751 TI - [Homosexuality and "reorientation therapy"--again]. PMID- 15815752 TI - Plasma concentration of atrial natriuretic peptide is related to the duration of atrial fibrillation in patients with advanced heart failure. AB - BACKGROUND: Plasma concentration of atrial natriuretic peptide (ANP) is elevated in patients with atrial fibrillation (AF) and in patients with chronic heart failure (CHF).Aim. To assess ANP level in patients with permanent AF and advanced CHF. METHODS: The study group consisted of 41 patients (27 males, mean age 62+/-8 years) with AF of a mean duration of 8.8 months. Twenty six (63%) patients were in NYHA class II, and 15 (37%) - in NYHA class III or IV. All patients underwent clinical and echocardiographic evaluation as well as ANP plasma concentration assessment. Multiple regression analysis was used to identify factors which determine ANP plasma concentration. RESULTS: Mean ANP plasma concentration was 52.4+/-22.7 pg/ml in the whole study group; 38.6+/-10.8 pg/ml in NYHA class II patients and 74.9+/-18.7 pg/ml in NYHA class III-IV subjects (p<0.0001). Among echocardiographic parameters, patients with NYHA class III or IV had significantly lower left ventricular ejection fraction and greater left atrial volume than patients with NYHA class II (32% versus 56%, p<0.0001 and 101.0+/ 23.8 cm(3) versus 83.4+/-16.1 cm(3), p<0.006, respectively). Multiple regression analysis revealed a significant negative correlation between AF duration and ANP level (p=0.0013) in a group of patients with NYHA class III or IV and identified AF duration as an independent predictor of ANP plasma concentration in this group of patients. CONCLUSIONS: ANP plasma concentration in patients with persistent AF and advanced CHF is determined by AF duration - the longer the AF duration the lower the ANP level. PMID- 15815754 TI - Coronary atherosclerosis in patients with acquired valvular disease. AB - BACKGROUND: Coronary atherosclerosis often coexists with acquired valvular disorders. There is growing evidence in literature that these two conditions may have common aetiology. AIM: To assess the incidence of coronary atherosclerosis in patients with acquired valvular disorders and to compare clinical parameters as well as the prevalence of risk factors between patients with aortic and mitral valve diseases. METHODS: The study group consisted of 155 patients (101 males, 54 females, mean age 58.2+/-9.7 years) with acquired valvular disorder who between 2000 and 2002 underwent invasive cardiac evaluation in our department prior to planned cardiac surgery. Aortic stenosis was detected in 74 patients, aortic insufficiency -- in 26, mitral stenosis -- in 33, and mitral regurgitation -- in 14 subjects. All patients underwent clinical evaluation, echocardiography, coronary angiography and laboratory tests. RESULTS: Patients with aortic stenosis had similar prevalence of coronary atherosclerosis to patients with aortic insufficiency, and patients with mitral stenosis -- to patients with mitral regurgitation. When the two groups -- patients with aortic valve disease and patients with mitral valve disease were compared, significant coronary lesions were more often detected in patients with aortic valve disease (36% vs 12.8%, p<0.05). Also, patients with aortic valve disorder were older, predominantly of male gender, had more often angina but less often heart failure, and had higher total cholesterol level than patients with mitral valve disease. CONCLUSIONS: Significant coronary lesions are more frequently encountered in patients with aortic valve disorder than in those with mitral valve disease. A high prevalence of atherosclerotic risk factors in patients with aortic valve disease may suggest that this condition has similar aetiology to that of coronary artery disease. PMID- 15815753 TI - Percutaneous thrombectomy with the RESCUE system in acute myocardial infarction. AB - BACKGROUND: Percutaneous coronary interventions (PCI) in acute myocardial infarction with ST segment elevation (STEMI) are associated with distal coronary embolisation. It may be speculated that percutaneous thrombectomy preceding stent implantation may prevent coronary microcirculation from embolisation. AIM: To assess safety and efficacy of percutaneous thrombectomy in patients with STEMI. METHODS: Seventy two patients with STEMI were randomised to PCI with stent implantation alone (n=32) or percutaneous thrombectomy with the RESCUE system, followed by stent implantation (n=40). Coronary flow in infarct related artery before and after the procedure was assessed using TIMI scale and corrected TIMI frame count - cTFC. Myocardial blood flow was measured using TIMI myocardial perfusion grade - tMPG. The degree of ST segment resolution 60 min after PCI was also assessed. Left ventricular ejection fraction (LVEF) was measured in hospital and three months later. RESULTS: The two groups did not differ with respect to the time from the onset of symptoms to the procedure (236+/-162 min vs 258+/-198 min, NS) or the baseline TIMI, cTFC and tMPG values. An effective thrombectomy procedure was performed in 35 (87%) patients from group B. After the procedure, the number of patients with TIMI 3 grade as well as cTFC values and the proportion of patients with tMPG 3 were similar in both groups (86% vs 85%, NS; 19 vs 21, NS; and 38% vs 54%, NS). The sum of ST segment elevations after the procedure was significantly greater in patients who underwent PCI only compared with patients who had thrombectomy and PCI (6.8+/-5.2 mm vs 3.6+/-2.9 mm, p=0.004). Complete normalisation of ST segment was achieved in 68% of patients treated with thrombectomy and PCI compared with 25% of patients who had PCI only (p=0.005). CK-MB peak values occurred significantly earlier in patients treated with thrombectomy (92.1% vs 66.7% up to 360 min, p=0.01). After 3 months of follow-up, LVEF tended to be greater in patients treated with thrombectomy and PCI than in those who underwent PCI only (55.3+/-14.7% vs 60.3+/-9.2%, NS). CONCLUSIONS: Thrombectomy with the RESCUE system in patients with STEMI is safe and effectively restores patency of infarct related artery. Thrombectomy better improves myocardial perfusion than standard PCI. PMID- 15815755 TI - Effects of social intervention on detection and efficacy of treatment for arterial hypertension. Main results of the Polish Four Cities Programme. AB - BACKGROUND: Cardiovascular diseases are the main cause of death in the adult Polish population. Beside lipid disorders and cigarette smoking, hypertension represents the most important risk factor leading to cardiovascular complications. Representative studies conducted in Poland in 1994-2002 showed that in 2002 the number of respondents in the survey who stated they knew their own blood pressure values dropped by 3.5 million, compared with 1994. This decrease was predominantly seen in small towns and in the countryside. Preventive programmes should therefore be addressed mainly to the most vulnerable communities. Modern methods of social marketing may play a substantial role in the creation of a healthy lifestyle. AIM: The aim of the Polish Four Cities Programme (PP4M), conducted in 2000-2001, was to develop the most effective methods of detection of and improvement in treatment for hypertension among the residents of small towns and rural areas. One of the programme tasks was to compare the effectiveness of a standard medical screening intervention with a similar approach combined with the use of social marketing methods.Methods. The programme was conducted by an interdisciplinary team in three small Polish towns - Kartuzy, Oborniki Wlkp. and Braniewo, as well as in one of the districts of a large city Lodz -- Olechow. Medical intervention combined with social marketing (community intervention) took place in Oborniki Wlkp. whereas the residents of Kartuzy and Lodz were subjected only to the traditional medical intervention. Braniewo served as a control location -- neither medical nor community intervention was implemented. Community intervention with elements of social marketing consisted of a three-month, intensive education and information campaign, initiated four weeks prior to the start of medical intervention. Epidemiological situation was assessed in all the four cities before and after the completion of the preventive interventions (screening), using representative surveys, with the objective to assess the changes in the awareness of one's own blood pressure values, detection of hypertension and knowledge concerning cardiovascular risk factors.Results. In two survey locations -- Kartuzy and Lodz awareness of one's own blood pressure values after the medical intervention did not significantly change (61% and 67.6% at baseline versus 62.1% and 71.6% after the intervention, respectively). In contrast, social marketing activities conducted in Oborniki significantly increased this parameter from 61.5% to 79.8% (p<0.01). While medical intervention did not change the proportion of non diagnosed hypertension in a small town (a non-significant decrease from 49% to 45% in Kartuzy), its effect in a large city was clearly visible (a decrease from 46% to 28% in Lodz). In Oborniki Wlkp. (medical intervention combined with social marketing) the effects were the most noticeable -- a reduction from 50% to 27% was achieved. The efficacy of hypertension treatment at baseline was low (4.7% in Kartuzy, 6.6% in Oborniki, and 6.5% in Lodz), but it then improved significantly (a twofold increase in Kartuzy and Oborniki, and more than twofold increase in Lodz). When the target value of blood pressure was set at 160/95 mmHg, the highest efficacy of hypotensive therapy was observed directly after the completion of medical and community intervention in Oborniki (an almost twofold increase in treatment efficacy). CONCLUSIONS: 1. Medical intervention combined with a community intervention and marketing campaign leads to a statistically significant improvement in self-awareness of blood pressure values among residents of small towns. 2. Medical intervention combined with community intervention brings the detection rate of hypertension in small towns up to the level observed in large cities. 3. Medical intervention, especially when combined with community intervention, improves the efficacy of the treatment of hypertension, regardless of the size of agglomeration. PMID- 15815756 TI - Clinical presentation and pharmacological therapy in patients with cardiogenic pulmonary oedema. AB - BACKGROUND: Cardiogenic pulmonary oedema is a typical clinical presentation of acute heart failure and is associated with a poor outcome. Therapeutic strategies in patients with this condition have not yet been uniformly developed and are based on expert consensus rather than results of randomised studies. The underlying aetiology seems to be one of the most important factors influencing therapy. AIM: To evaluate treatment of patients with acute cardiogenic pulmonary oedema. METHODS: The analysed group consisted of 276 consecutive patients who were treated due to acute cardiogenic pulmonary oedema in two cardiac centres in the city of Lodz between 1998 and 2000. Clinical characteristics, aetiology and therapy were retrospectively analysed based on medical records. RESULTS: Fifty nine (21%) patients died in hospital whereas the remaining 218 subjects were discharged. Etiologic Results. Fifty nine (21%) patients died in hospital whereas the remaining 218 subjects were discharged. Etiologic factors included a marked elevation of blood pressure (BP) (29%), acute myocardial infarction (MI) (25%), unstable angina (16%), decompensation of valvular disorder (9%), respiratory tract infection (8%) and paroxysmal atrial fibrillation (4%). The highest in hospital mortality (35%) was associated with acute MI, and the lowest (6%) - with BP elevation. The most frequently used agents during in-hospital stay were diuretics (89%), followed by antibiotics (84%) and acetylsalicylic acid (81%). Compared with survivors, patients who died during hospital stay were significantly more frequently treated with catecholamines (59% vs 7%, p<0.00001), corticosteroids (71% vs 15%, p<0.00001) and morphine (62% vs 41%, p<0.0015) whereas angiotensin converting enzyme inhibitors, intravenous nitroglycerine, beta-blockers and acetylsalicylic acid were less frequently used in deceased patients (34% vs 89%, p<0.00001; 52% vs 78%, p<0.00001; 14% vs 37%, p<0.001; and 64% vs 86%, p<0.0003, respectively). CONCLUSIONS: In spite of intensive pharmacotherapy, cardiogenic pulmonary oedema is still associated with a high in hospital mortality. Therapy should be tailored according to aetiology. Differences in pharmacological treatment between survivors and deceased patients depict better prognosis in those who have on admission normal or elevated blood pressure which enables an early administration of vasodilators and beta-blockers. PMID- 15815757 TI - [Acute coronary syndrome or perimyocarditis?--case report]. AB - Acute coronary syndrome or perimyocarditis? A case report. A case of a 25-year old male with a severe chest pain associated with ST segment changes, elevated serum C-reactive protein, myocardial necrosis markers and normal epicardial coronary arteries is presented. The patient recently recovered from upper respiratory infection. Differential diagnosis between acute coronary syndrome and perimyocarditis is discussed. PMID- 15815758 TI - [Electrical remodeling in a patient with biventricular pacemaker]. AB - Electrical remodelling in a patient with biventricular pacemaker - a case report. A case of a 70-year-old patient with dilated cardiomyopathy is presented. The patient underwent biventricular pacemaker implantation and improved markedly. Indications for resynchronisation therapy are discussed. PMID- 15815759 TI - [Cardiac myxoma attached to anterior leaflet--case report]. AB - Cardiac myxoma attached to anterior mitral leaflet - a case report. A case of a 64-year-old patient with a history of stroke and myocardial infarction is presented. Echocardiography revealed cardiac myxoma originating from anterior mitral leaflet. The patient underwent successful surgery. Differential diagnosis of cardiac myxoma is presented. PMID- 15815760 TI - [Persistent left superior vena cava]. AB - Persistent left superior vena cava -- a case report. A case of a 72-year-old female admitted for pacemaker implantation is presented. During the procedure, persistent left superior vena cava was found. Pacemaker was inserted via right subclavicular vein. PMID- 15815761 TI - [Value of medical history and ECG stress testing in detection of restenosis following successful balloon coronary angioplasty]. PMID- 15815762 TI - [Evidence-based medicine. Interpretation of results is also important]. PMID- 15815764 TI - [Patients I will always remember]. PMID- 15815763 TI - [Cardiologic health care in Poland--report from cardiology consultants in the year 2004]. PMID- 15815765 TI - [Letter from prof. Andrzej Cieslinski to the editor of Kardiologia Polska prof. Leszek Ceremuzynski]. PMID- 15815767 TI - [Carbon monoxide poisoning or acute myocardial infarction? Case report]. PMID- 15815768 TI - [Ostial stenosis of the left main stem coronary artery--a case report]. PMID- 15815769 TI - [Angiogram of the month--unstable angina without ST segment elevation]. PMID- 15815770 TI - [Ventricular tachycardia resistant to pharmacological and ablation therapy--a case report]. PMID- 15815771 TI - [New European guidelines on cardiovascular disease prevention in relation to the situation in Poland]. PMID- 15815773 TI - Coronary flow velocity reserve is diminished in hypertensive left ventricular hypertrophy. AB - BACKGROUND: Dipyridamole stress transesophageal echocardiography (STEE) is a feasible method for the evaluation of coronary flow velocity reserve (CFR). AIM: The aim of the present study was to investigate CFR in hypertensive patients with or without left ventricular hypertrophy (LVH). METHODS: The study comprised 73 patients with a negative coronary angiogram (29 men and 44 women). Three different groups were compared: normotensive patients, hypertensive patients without LVH and hypertensive patients with LVH. RESULTS: CFR was significantly decreased in patients with hypertension with LVH as compared to normotensive cases (2.19+/-0.50 vs 2.71+/-1.10; p<0.05). CFR of hypertensive patients without LVH was only slightly reduced as compared to normotensive cases (2.44+/-0.81 vs 2.71+/-1.10; p=ns). In hypertensive patients with LVH, the LV mass and LV mass index were inversely related to CFR (r = -0.481 and -0.477, p<0.05, respectively). CONCLUSIONS: CFR is diminished in patients with hypertension. The degree of CFR reduction is related to the extent of LVH. PMID- 15815774 TI - Long-term outcome of conservatively treated patients with borderline coronary lesions--role of the fractional flow reserve measurement. AB - BACKGROUND: Coronary fractional flow reserve (FFR) has been recommended as one of the functional methods which can be used to establish indications for revascularisation in patients with borderline coronary lesions. AIM: To assess long-term outcome of patients with borderline coronary lesions in whom the decision to implement conservative treatment was based on the results of FFR. METHODS: The study group consisted of 41 patients (13 females, mean age 61+/-9 years) who had CCS class II angina, an isolated borderline (40-70%) coronary lesion and FFR >0.75. All patients received 40 mg of simvastatin, angiotensin converting enzyme inhibitor and aspirin. The follow-up duration ranged from 7 to 32 months (mean 15.2+/-7.1, median 14.5 months). The analysed end-points included cardiac death, myocardial infarction (MI) (with or without ST segment elevation) and revascularisation of the target coronary artery. RESULTS: The mean FFR value in the whole study group was 0.91+/-0.05. Three (7%) patients had complications during follow-up: one patient developed MI, and two had coronary artery stenosis progression, requiring angioplasty. These patients had similar clinical and angiographic characteristics as well as FFR values as patients without complications. Among patients with uneventful outcome, in 33 (87%) anginal symptoms improved whereas in the remaining 5 (13%) patients CCS class did not change. Patients with angina alleviation were older and had higher FFR values than patients without improvement (63+/-8 vs 48+/-6 years, p=0.0005; and 0.92+/ 0.05 vs 0.86+/-0.08, p=0.04, respectively). CONCLUSIONS: In patients with borderline coronary lesions and stable angina, angioplasty can be delayed on the basis of the FFR results. This approach is safe and is associated with an asymptomatic long-term follow-up in more than 90% of patients. PMID- 15815775 TI - Aspirin resistance in ischaemic heart disease. AB - BACKGROUND: In spite of the usage of acetylsalicylic acid (aspirin) in the secondary prevention of ischaemic heart disease (IHD), new thrombo-embolic events occur in more than half of patients. Aspirin resistance may be partially responsible for this phenomenon. AIM: To assess the prevalence of aspirin resistance in patients with IHD and to correlate this phenomenon with the progression of atherosclerosis, concomitant diseases and other medication. METHODS: The study group consisted of 205 patients (mean age 65.8 years, 95 females) with stable angina, recent coronary angiography and positive result of non-invasive stress tests, treated with 75 mg of aspirin for at least one week. Platelet aggregation was measured using the optical aggregation method. Aspirin resistance was defined as a mean collagen and ATP-induced platelet aggregation >70%. RESULTS: Aspirin resistance was found in 41 (20%) patients and was significantly associated with previous coronary artery bypass grafting (CABG) (p<0.01) and three-vessel disease (p<0.05). Previous CABG was the only independent risk factor for the presence of aspirin resistance (OR 5.6; 95% CI 2.0-15.4; p<0.01). CONCLUSIONS: Aspirin resistance is present in 20% of patients with stable angina. Previous CABG is an independent risk factor of this phenomenon. PMID- 15815776 TI - Effect of leukocyte-depleted blood cardioplegia on the early outcome in patients with preserved left ventricular function undergoing surgical revascularisation. AB - BACKGROUND: It has been shown that leukocytes play one of the key roles in the myocardial reperfusion injury. AIM: To examine the effects of cardiac protection with leukocyte-depleted blood cardioplegia on the early outcome of patients with preserved left ventricular function who undergo surgical revascularisation. METHODS: The study group consisted of 58 patients with coronary artery disease (CAD) undergoing coronary artery bypass grafting (CABG) who were randomised to receive leukocyte-depleted blood cardioplegia (leukocyte filter Pall BC1B) (group A, n=29) or to receive standard blood cardioplegia (group B, n=29). Peri operative mortality and morbidity as well as haemodynamical and biochemical parameters were compared between these two groups. RESULTS: No early death occurred. There were no statistical differences in clinical data between the groups. Only cardiac index measured 24 hours after declamping of aorta was significantly higher in group A than in group B (3.6+/-0.6 l/min/m(2) vs 2.95+/ 0.45 l/min/m(2), p<0.05). Group B showed significant higher release of creatine kinase (CK) 6 and 12 hours, and CK-MB 6, 12, and 24 hours after unclumping the aorta whereas troponin I level was similar in both groups. CONCLUSIONS: The use of leukocyte-depleted blood cardioplegia during elective CABG did not improve the early outcome. PMID- 15815777 TI - Iatrogenic cardiac arrhythmias following transcatheter or surgical closure of atrial septal defect in children. AB - BACKGROUND: Invasive procedures involving the atria may promote the development of iatrogenic cardiac arrhythmias. AIM: To analyse the prevalence of cardiac arrhythmias following transcatheter or cardiosurgical closure of the secundum type atrial septal defect (ASD). METHODS: The study group consisted of 91 patients, aged 2-18 years with haemodynamically significant ASD who underwent surgical (n=44) or transcatheter (Amplatzer occluder) (n=47) closure of ASD. Standard ECG and Holter ECG recordings, obtained before and after the procedure, were analysed. The follow-up duration ranged from 2.5 to 5.5 years. Cardiac arrhythmias were divided into benign or significant (requiring pharmacological therapy), early or late, and transient or permanent. RESULTS: Cardiac arrhythmias were detected in 16 (36%) patients who underwent surgery compared with 1 (2.1%) patient who underwent transcatheter ASD closure (p<0.05). In surgically treated patients, arrhythmias were benign in 9 patients, significant in 7 children, early in 15 subjects, late in one patient, transient in 13 children and permanent in 3 subjects. One patient, who underwent transcatheter ASD closure, developed paroxysmal supraventricular tachycardia one day after the procedure, successfully terminated with verapamil. CONCLUSIONS: Transcatheter closure of ASD is associated with a lower risk of procedure-related arrhythmias than surgical treatment. However, longer follow-up in patients treated with transcatheter procedure is needed in order to draw definite conclusions. PMID- 15815778 TI - [Profound thrombocytopenia after abciximab administration in acute myocardial infarction and stent thrombosis following thrombocytopenia remission--a case report]. AB - Profound thrombocytopenia after abciximab administration in acute myocardial infarction and stent thrombosis. This article presents a case of a 62-years-old man with acute anterior myocardial infarction, treated with PCI and stent implantation, in whom profound acute thrombocytopenia was observed after abciximab administration. Nadir platelet count was 6 G/L (before treatment: 250 G/L). Pseudothrombocytopenia was excluded. The remaining antiplatelet drugs (heparin, ASA, clopidogrel) were discontinued. There were no symptoms of bleeding, but next morning (platelet count: 14 G/L) a gross hematoma at femoral puncture site was observed. The patient received 5 U transfusion of platelets. On the 4th day, when the platelet count reached 64 G/L, he was started again on ASA (150 mg) and clopidogrel (75 mg). On the 7th day (platelet count: 138 G/L) he developed anterior ischemia and stent reocclusion was diagnosed. After p.o. clopidogrel (300 mg), balloon PCI with i.c. heparin was performed and ischemia symptoms subsided. The platelet value before the patient's discharge, on subsequent therapy with ASA and clopidogrel, increased to 300 G/L. A review of current literature on this topic is provided. PMID- 15815779 TI - [Peripartum cardiomyopathy--a case report]. AB - Peripartum cardiomyopathy -- a case report. A case of a 32-year old woman, previously healthy, with heart failure symptoms occurring during third pregnancy, is described. In spite of standard pharmacological treatment, her condition worsened and the pregnancy had to be terminated at 28 hbd by cesarean section. The patient's condition improved and three months later normal left ventricular function as well as good exercise tolerance were observed. PMID- 15815780 TI - [Gastroesophageal acid reflux as a causative factor of paroxysmal atrial fibrillation]. AB - We present a case of a patient with very frequent episodes of paroxysmal atrial fibrillation (AF) who, besides hypertension and well-controlled post-amiodarone thyreotoxicosis, had no other potential causes of AF. Because of symptoms of gastroesophageal acid reflux, the patient was treated with omeprazole which significantly reduced the frequency of AF episodes. Next, the patient underwent anti-reflux surgery and has now only sporadic recurrences of AF. Our observations suggest that gastroesophageal acid reflux may be a triggering factor of AF. PMID- 15815781 TI - [Common origin of the right and left coronary artery from the right sinus of Valsalva detected by transthoracic echocardiography--case report]. AB - Common origin of the right and left coronary artery from the right sinus of Valsalva detected by transthoracic. A case of a 69-year old female with a long lasting history of angina, is presented. During hospitalisation due to angina aggravation, transthoracic echocardiography was performed and revealed anomalous coronary arteries. The finding of a common origin of the right and left coronary artery from the right sinus of Valsalva was confirmed by coronary angiography. PMID- 15815782 TI - [Modified albumin as a marker of myocardial ischemia]. PMID- 15815783 TI - [Hemodynamic processes during cardiopulmonary resuscitation. Controversial problem?]. PMID- 15815788 TI - [Acute pancreatitis mimicking acute myocardial infarction or vice versa? An EKG case report]. PMID- 15815789 TI - [Two cases of inferior-posterior atrial septal defects--comprehensive echocardiographic evaluation]. PMID- 15815790 TI - [Thermography and ultrasonography in diagnosis of patients with acute coronary syndrome]. PMID- 15815791 TI - [RF ablation of a permanent atrial tachycardia in a pregnant woman without X-Ray use in the 24th week of pregnancy]. PMID- 15815792 TI - [Aspirin resistance--pharmacological mechanisms and clinical implications]. PMID- 15815793 TI - Body mass index and risk of death in middle-aged men and women in Poland. Results of POL-MONICA cohort study. AB - BACKGROUND: Overweight and obesity have been considered to be the causes of serious health consequences. So far, there have been no studies in Poland to analyse the relationship between Body Mass Index (BMI) and mortality. AIM: To determine the relationship between BMI and the mortality risk due to all causes and due to cardiovascular diseases (CVD) in the middle-aged population of Poland. METHODS: The study group consisted of men and women aged 35-64, residents of two districts of Warsaw (Praga Polnoc and Praga Poludnie) and the former Tarnobrzeg Voivodship who were selected at random for POL-MONICA Project cross-sectional studies in 1983-1994. Risk assessment of death due to all causes and due to CVD was performed using the Cox proportional hazards method. Persons with BMI of 22 23.9 were adopted as reference group. RESULTS: Sample size consisted of 5,281 men and 5,691 women. Prospective observation was carried out from 5 to 15 years, mean duration 10.7 years. Overall, the observation comprised 117,839 person-years. During the study, 914 men died (including 448 due to CVD) as well as 430 women (including 160 due to CVD). After adjusting for age, place of residence (Warsaw vs. the Tarnobrzeg Voivodship), risk factors (hypercholesterolemia, arterial hypertension and smoking) and self-assessment of health, increased risk of all cause death was found in men with BMI lower than 20 kg/m(2) (relative risk 2.27), with BMI of 32-34.9 kg/m(2) (relative risk 1.41), with BMI > or =35 kg/m(2) (relative risk 1.73), and in women whose BMI was lower than 20 kg/m(2) (relative risk 1.66). Increased risk of CVD death was found in men with BMI lower than 20 kg/m(2) (relative risk 1.97), with BMI of 32-34.9 kg/m(2) (relative risk 1.66), with BMI > or =35 kg/m(2) (relative risk 2.06), and in women with BMI > or =35.0 kg/m(2) (relative risk 2.02). CONCLUSIONS: The POL-MONICA study carried out in the middle-aged Polish population did not confirm a relationship between overweight (BMI from 25 to 29.9 kg/m(2)) and risk of death due to all causes or due to CVD. PMID- 15815794 TI - Inter-individual variability in response to clopidogrel in patients with coronary artery disease. AB - BACKGROUND: Clopidogrel, especially when combined with aspirin, reduces the rate of ischaemic events in patients with coronary artery disease (CAD). There are scare data in literature on the inter-individual variability in response to clopidogrel. AIM: To assess the incidence of clopidogrel resistance using rapid whole blood platelet function assessment, and to examine the possibility of early identification of non-responders. METHODS: In 31 consecutive patients with stable angina treated with aspirin, the degree of platelet aggregation inhibition (DPAI) in the whole blood was assessed at baseline and 3, 6, 12 as well as 24 hours after administration of loading dose of clopidogrel (300 mg). Response to clopidogrel was measured by calculating the absolute difference between the baseline DPAI and DPAI obtained at the investigated time-points (DPAI). RESULTS: After 24 hours from clopidogrel administration, seven (22.6%) patients were identified as non-responders (DPAI < or =10%). Demographic and clinical variables as well as baseline DPAI were similar in responders and non-responders (DPAI: 5.8+/-3.7% vs 7.1+/-5.3%, p=NS). Out of the patients who were found to be resistant to clopidogrel at the six-hour time-point, 87.5% remained resistant to this agent 24 hours after drug administration. DPAI calculated at the 24-hour time-point highly correlated with the six-hour DPAI (r=0.74). No differences in the rate of ischaemic or bleeding complications between responders and non responders were noted. CONCLUSIONS: The assessment of the degree of platelet aggregation inhibition allows early (six hours from the initiation of treatment) identification of patients who are resistant to clopidogrel. The method of the rapid whole blood platelet function assessment is feasible in every-day clinical practice. PMID- 15815795 TI - Does saddle embolism influence short-term prognosis in patients with acute pulmonary embolism? AB - BACKGROUND: In some patients with acute pulmonary embolism (APE) thrombi may lodge at the levels of the bifurcation of pulmonary trunk and extend into both main pulmonary arteries, forming so-called saddle embolism (SE). AIM: To assess the incidence of SE and whether it is associated with an increased risk of complicated clinical course. METHODS: We studied 150 consecutive patients (94 females, 56 males) aged 63.6+/-16.7 years with APE confirmed with contrast enhanced spiral computed tomography or transesophageal echocardiography. RESULTS: SE was detected in 22 (14.7%) patients. Mean age (SE vs N-SE) was 64.3+/-17.4 vs 63.5+/-16.6 years, heart rate 100.8+/-14.1 beats/min vs 97.8+/-21.1 beats/min, systolic blood pressure 126.2+/-20.1 vs 127.1+/-23.3 mmHg and blood pulsoximetry 92 (68-98) vs 91 (30-98) % (all differences NS). In patients with SE, echocardiographic signs of the right ventricular overload, defined as right to left ventricular end - diastolic ratio >0.6 with right ventricular hypokinesia and/or maximal tricuspid peak systolic gradient >30 mmHg with shortened acceleration time of pulmonary ejection <80 ms, were more frequent (77.3% vs 51.6%, p=0.04), as was the mid-systolic deceleration of pulmonary ejection velocity (77.3% vs 49.2%, p=0.04). Mortality and complicated clinical course rates were similar in patients with SE or N-SE (mortality: 4.5% vs 13.3%, NS, and complicated clinical course: 34.4% vs 25.0%, NS). CONCLUSIONS: Saddle pulmonary embolism is frequent, especially in patients with echocardiographic signs of impaired pulmonary ejection pattern. Saddle embolism does not indicate unfavourable clinical outcome and probably should not influence treatment selection. PMID- 15815796 TI - Value of lead aVR in the detection of significant left main coronary artery stenosis in acute coronary syndrome. AB - BACKGROUND: Electrocardiographic lead aVR is usually ignored in patients with chest pain. ST segment elevation in aVR may have diagnostic value in patients with acute coronary syndrome (ACS) and significant stenosis or obstruction of the left main coronary artery (LMCAS), especially when accompanied by ST segment elevation in lead V(1). AIM: To asses the value of lead aVR and V1 for the detection of LMCAS in patients with ACS. METHODS: The study group consisted of 150 patients (mean age 60.6+/-9.5 years, range 33-78 years) with ACS, including 46 with LMCAS and 104 without LMCAS. ECG recordings obtained on admission were compared between these two groups. RESULTS: In patients with LMCAS, ST segment elevation in lead aVR was two times more frequent than in remaining patients (69.6% vs 34.6% p=0.0001) whereas there were no differences in lead V(1). Sensitivity of ST elevation in aVR in detection of LMCAS was 69.6%, specificity - 65.4%, positive predictive value - 47.1%, and negative predictive value - 82.9%. In patients with LMCAS, ST segment depression was significantly more often present in ECG leads other than aVR (45.6% vs 23.1% p<0.01). Patients with LMCAS more often had hypertension (95.6% vs 77.9% p<0.05) and three-vessel disease (78.3% vs 31.8%, p<0.0001). CONCLUSIONS: The assessment of lead aVR in patients with ACS may indicate LMCAS. Additional analysis of lead V(1) does not improve diagnostic accuracy. PMID- 15815797 TI - Effectiveness of radiofrequency catheter ablation of right ventricular outflow tract tachycardia using the CARTO system. AB - BACKGROUND: Ventricular ectopy or ventricular tachycardia (VT) originating from the right ventricular outflow tract (RVOT) are the most common forms of arrhythmias in patients with structurally normal heart. Pharmacological treatment is effective in no more than 50% of patients, whereas radio-frequency catheter ablation (RFCA) offers a much higher success rate. AIM: To assess early and late outcome in patients with RVOT arrhythmias treated with RFCA combined with electro anatomical mapping system (CARTO). METHODS: The study group consisted of 34 consecutive patients (mean age 38.8+/-12.0 years, range 21-52 years, 11 males, 23 females) with symptomatic arrhythmias originating from RVOT, who underwent RFCA in our department between December 2001 to July 2003. RFCA was performed with the use of the CARTO system. The power of RF current was set at 40 Watts, duration -- 90 seconds, and maximal temperature -- 55 degrees C. In order to assess short- and long-term RFCA efficacy, a 24-hour Holter ECG monitoring was performed before RFCA and shortly after the procedure as well as one and three months afterwards. Effective RFCA was defined as the reduction of ventricular ectopy <1000 / 24 hours in Holter monitoring performed just after the procedure. RESULTS: Holter ECG monitoring performed after RFCA showed that the procedure was effective in 30 (88.2%) patients. In the remaining four patients no significant reduction in the frequency of ventricular ectopy was noted, however, no complex ventricular arrhythmias were present. In none of the patients neither early nor late complications were observed. The mean follow-up duration was 15.6 months (range 5 26 months). During Holter ECG monitoring performed one and three months after RFCA, a recurrence of frequent ventricular ectopy (7139 beats / 24 hours) was found in one patient, however, without complex arrhythmias. The remaining patients, in whom RFCA was found to be effective at Holter ECG monitoring performed just after the procedure, continue to be free from arrhythmia and do not require antiarrhythmic agents. CONCLUSIONS: RFCA with the use of the CARTO system is effective and safe in the treatment of arrhythmias originating from RVOT. PMID- 15815798 TI - [Acute coronary syndrome without ST segment elevation in a patient with familial hypertrophic cardiomyopathy--a case report]. AB - Acute coronary syndrome without ST segment elevation in a patient with familial hypertrophic cardiomyopathy. A case of a 46-year-old male with hypertrophic cardiomyopathy and cardioverter-defibrillator implanted due to a history of syncopal sustained ventricular tachycardia, is presented. The patient had undergone coronary angiography two years before current hospitalisation which showed normal coronary arteries. This time the patient was admitted to the hospital due to a typical chest pain. ECG showed predominantly paced QRS complexes and negative T waves in V2-V6 which were present on ECG recorded a few months earlier. Troponin I plasma concentration was significantly elevated. Coronary angiography revealed critical stenosis of the left anterior descending coronary artery which was successfully treated with angioplasty and stent implantation. Diagnosis and treatment of patients with hypertrophic cardiomyopathy are discussed. PMID- 15815799 TI - [Single coronary artery in a patient with acute myocardial infarction and supraventricular tachycardia--a case report]. AB - A case of a 66-year-old patient with a history of paroxysmal supraventricular tachycardia, hypertension and chronic obturatory pulmonary disease is presented. The patient was admitted to the hospital due to acute myocardial infarction. Coronary angiography revealed a single coronary artery originating from the right sinus of Valsalva without any significant lesions. Literature concerning this rare coronary anomaly is discussed. PMID- 15815800 TI - [Resistance to activated protein C in a patient with multiple myocardial infarctions and stroke--a case report]. AB - A case of a 49-year-old female with a history of two myocardial infarctions (MI) and ischaemic stroke is presented. The patient was admitted to the hospital due to a third acute MI. Laboratory investigations revealed resistance to activated protein C due to factor V Leiden mutation. Diagnosis and treatment of patients with this condition are discussed. PMID- 15815801 TI - [Evaluation of the coronary risk: is it enough to asses ApoB/ApoA1 ratio only or is plasma lipoprotein concentration still necessary?]. PMID- 15815802 TI - [Procedures of invasive cardiology performed in ambulatory care]. PMID- 15815806 TI - [An ECG resembling acute myocardial infarction]. PMID- 15815807 TI - [Right atrial thrombi in a patient with congestive cardiomyopathy]. PMID- 15815808 TI - [Angiogram of the month]. PMID- 15815809 TI - [Drivers from the vein of Marshall, left pulmonary veins, and inferior right atrial isthmus for mosaic of incessant left, right and combined atrial tachyarrhythmias--role of selective mapping and RF ablation]. PMID- 15815810 TI - [Pulmonary endarterectomy in chronic thromboembolic pulmonary hypertension]. PMID- 15815811 TI - Evaluating effects of preservative-containing saline solution on pain perception during botulinum toxin type-a injections at different locations: a prospective, single-blinded, randomized controlled trial. AB - Pain is a side effect of botulinum toxin type-A (BTX-A) injections. The efficiency of the preservative-containing saline solution used as the "dilution solution" for controlling the pain felt during multiple injections in different areas has been investigated by a prospective, randomized, single-blinded, controlled study. A total of 93 patients were divided into three groups for the study. All the patients were given BTX-A. Of these 93 patients, 60 received injections in the upper face, 15 in the neck, and 18 in the axillary regions. The visual analog scale (VAS), a single-dimension pain intensity rating scale, was used to evaluate pain perception. The average VAS values for pain sensation experienced by the groups were 1.2 of 10 points for the upper face area in the experimental group (n = 60) and 4.5 point for the control group. In the neck region (n = 15), the respective scores were 0.6 in the experimental group and 3.9 in the control group. Finally, in the axillary region (n = 18), the respective values were 0.9 and 5.1. The authors conclude that the preservative-containing saline solution significantly decreased pain perception during BTX-A injections (p = 0.000). PMID- 15815812 TI - Experience of treating gunshot wounds of large vessels in Afghanistan. AB - The problem of treating injuries of large vessels in times of peace has been sufficiently well developed both in theoretical and practical aspects. In times of war, however, due to the large numbers of the wounded, a shortage of expert angiosurgeons, and multiple gunshot wounds, many theoretical tenets lose their academic regularity. The present study is based on the experience of treating 302 patients wounded during the Afghanistan war (1981-1985). Most of the injuries were to the vessels of the extremities. The gunshot wounds were complex. They included extensive destruction of tissue in the damaged segments of the extremities with simultaneous damage of large veins (42.0% of the cases), nerves (45.5%), and bones (47.4%). Multiple-vessel injuries were encountered in 4% of the cases, with combined injuries comprising 17.5%. Most of the wounded (83.7%) were in a state of shock, with 6.4% in a terminal condition. The blood loss amounted to 15% to 65% of the total volume. Ninety percent of the wounded were admitted to hospitals in the first 6 hours. A two-stage method was used to treat 71 of the wounded. The method included temporary bypass of the injured arteries and veins while evacuating the wounded and during surgery. Two hundred ninety five wounded underwent vessel surgery. Out of the total number of injuries of arteries and veins, vessel sutures were used in 36.9% and 35.9% of the cases, vessel plastics in 41.4% and 7.1% of the cases, and vessel ligation in 21.7% and 60.0% of the cases, respectively. Surgery was completed by fixing the bone fragments externally with the help of special devices using the Ilizarov method. Amputation was performed in 13.9% of the cases. In 7.3% of the cases amputation was performed according to primary indications (no reconstructive surgery attempted on the vessels). In 6.6% of the cases extremities were amputated during the early postoperative period as a result of vessel thrombosis and an increase of tissue ischemia. The mortality rate after vessel surgery was 5.3%. We believe that for patients with gunshot wounds involving vessel injuries, early one-time reconstruction of the destroyed anatomical structures should be performed. PMID- 15815813 TI - Landstuhl Medical Center's role in the global war on terrorism. PMID- 15815814 TI - Approach to postgraduate international education: an experience with professor Norman Rich. PMID- 15815815 TI - Pathophysiology of perforators in primary chronic venous insufficiency. PMID- 15815816 TI - Postgraduate education in emerging surgical technologies. PMID- 15815818 TI - Some aspects of road traffic injuries. PMID- 15815817 TI - Early research in the Department of Surgery, Uniformed Services University of the Health Sciences. PMID- 15815819 TI - The military medical school of Mexico: a tradition of excellence. AB - It is a historical fact that warfare and surgery have been linked together as far back as military history has been recorded. In the 18th century, the tendency of most armies to dismiss their medical services at the end of every major conflict resulted in higher mortality at the beginning of the next war. This became evident in the French and British Armies during the Battle of Waterloo. These countries went to great efforts to mobilize their civilian reserve physicians, only to discover that more than half of the medical personnel declined to serve. The scarcity of physicians and the inexperience of those caring for the wounded resulted in a high casualty rate. The current armed conflicts throughout the world with their high number of victims are living evidence of the need for preparedness of the military medical personnel. In this article, we review the systems of military medical education in several countries, and offer the example of the Escuela Medico Militar (Military Medical School) of Mexico, a prestigious source of military medical physicians for the Mexican armed forces. PMID- 15815821 TI - One surgeon's perspective. PMID- 15815820 TI - Cardiopulmonary assessment: is improvement needed? AB - Clinical parameters alone have repeatedly been proven unreliable in assessing cardiopulmonary status, especially in hemodynamically unstable patients. To learn if we had a diagnostic problem in our hospital, we compared physician assessment of cardiac index (CI) and thoracic fluid content (TFC) to values obtained using impedance cardiography (ICG). We selected the newest available ICG monitor, the BioZ, which employs this noninvasive technology. For CI measurements we have shown it to be equivalent to thermodilution and to be more reproducible (variability: 6.3% vs. 24.7%). Physician assessment of CI and TFC (high, normal, or low) was compared to the BioZ monitor's results in 186 patients, considered to be hemodynamically unstable, from the emergency room, the intensive care units, and the floors. Normal values were defined for CI (2.5-4.2 L/min m(2)) and for TFC (males: 30-50 kohm(-1) and females: 21-37 kohm(-1)). The concordance between physician assessment and the BioZ was 51% for CI with Kappa of 0.14 and 58% for TFC with Kappa of 0.19. Attendings did slightly better than the surgical residents with CI (52% vs. 48%) but slightly worse with TFC (57% vs. 61%). The potentially serious conditions of low CI and high TFC were misdiagnosed 42% and 46% of the time, respectively, by all physicians. Analysis of the data revealed that physician use of clinically available objective hemodynamic data, such as heart rate, blood pressure, and pulse pressure index, would not have been helpful. Furthermore, assistance from the pulmonary artery catheter (PAC) is often not available in our hospital, which has experienced a 90% decrease in its utilization over the past six years. Considering the increasing acuity of our aging patient population, accurate assessment of cardiopulmonary status is needed. The use of ICG could be a valuable addition to the physician's armamentarium. PMID- 15815822 TI - World War II in the Pacific and later at home. PMID- 15815823 TI - Hugh Williamson, M.D., LL.D. (1735-1819): soldier, surgeon, and founding father. PMID- 15815824 TI - Health care in Afghanistan. PMID- 15815825 TI - Dr. Norman M. Rich, the international military surgeon: his French connection. PMID- 15815826 TI - Norman M. Rich--the Walter Reed Vascular Surgery Fellowship. PMID- 15815827 TI - Abdominal war wounds--experiences from Red Cross field hospitals. AB - The traditional approach to abdominal war wounds consists of triage, eche-loned care, and mandatory laparotomy for penetrating abdominal injuries, and it remains valid in modern conventional wars with well-organized evacuation and surgical services. Expectant management of abdominal casualties can be considered under difficult circumstances with a high influx of patients exhausting the available resources. This can occur in regional conflicts associated with mass movements of people and with collapsed infrastructure. While always combined with adequate fluid resuscitation, antibiotic treatment, and other supportive care, the expectant approach in patients with penetrating abdominal injuries could be indicated for asymptomatic patients with multiple fragment wounds or for patients presenting several days post-injury in good condition. The focus of surgical resources and competence should be on the majority of patients with intestinal perforation only, who need surgery to save life--but not necessarily on an urgent basis--and who have a good chance of survival. The limited availability of blood products to correct blood loss and coagulation factor deficiencies, and the lack of sophisticated monitoring of hemodynamic variables that call into question the value of a damage-control approach for the most severely injured. Even if the bleeding could be temporarily controlled, the subsequent need for adequate resuscitation before returning the patient to the operating room could be difficult to achieve and would result in incompletely resuscitated patients being reoperated while acidotic, coagulopathic, and even hypothermic. Perhaps, in mass casualty situations these patients should be recognized during triage or at least early during operation, and aggressive surgery should be replaced with adequate expectant management with sedation and analgesics. PMID- 15815828 TI - Noninvasive vascular testing--a 35-year reflection. AB - Noninvasive vascular testing has made a major contribution to the care of vascular surgery patients. This article a reflection on a 35-year corroborative association with Doctor Norman Rich, during which many of the advances in noninvasive vascular testing have been witnessed and effected. I served my vascular surgery fellowship under Doctor Rich in 1967-68 at Walter Reed Army Hospital. The only noninvasive vascular equipment then was a pencil probe Doppler. The value of the Doppler in the management of peripheral vascular disease that year and in determining limb viability in combat surgery in Vietnam the following year was established, and both experiences were published. Later, Doctor Rich established the annual Military Vascular Surgery Meeting and was appointed as the first Chair of the Department of Surgery at the Uniformed Services University for Health Sciences (USU). I entered private practice in Central California in 1976, and ultrasonic imaging was developed which allowed noninvasive examination of the carotid arteries. I then developed a protocol to screen for the three silent, immediate causes of stroke, employing a "a quick carotid scan" for carotid artery disease, a lead II rhythm strip for atrial fibrillation, and blood pressure determination for hypertension so that these common causes of strokes could be recognized and treated, and potentially prevent the majority of strokes. My association with USU, Doctor Rich, and others involved proved instrumental in initiating implementation of stroke prevention screening. The structure established at USU provides a means of establishing the protocol nationally. Noninvasive vascular testing is an addition to Medical Science that has led to significant improvements in individual patient care and that has the potential of allowing a major reduction in death and disability from stroke and other vascular diseases. Throughout a 35-year collaborative association with Doctor Norman M. Rich, I have witnessed and developed many of these advances. These contributions to noninvasive vascular testing reflect the value of our collaboration. PMID- 15815829 TI - Carotid endarterectomy and stenting in management of extracranial carotid occlusive disease. PMID- 15815831 TI - Time for the Uniformed Services University of the Health Sciences (USUHS) to raise its sights. PMID- 15815832 TI - U.S.A.-Switzerland: a history of friendship. PMID- 15815830 TI - Wound healing enhancement: electroporation to address a classic problem of military medicine. AB - The major goal of wound healing biology is to determine how a wound can be induced to repair damaged tissue faster and more efficiently. Enhancement of dermal and epidermal regeneration is an extremely important goal for the treatment of many different types of wounds. Exogenous application of growth factors to the wound site has been shown to have potential to improve wound healing. Frequent applications of large amounts of growth factor have been required. This is because proteases in the wound quickly destroy peptide growth factor. Gene therapy has the potential to produce growth factors deep within the wound, where they can be effective as well as able to constantly replenish growth factor that is destroyed by peptidases. We have shown that application of plasmid DNA expression vectors directly into the wound is an inefficient modality. Electroporation, the application of an electrical field across cells to permeabilize the cell membrane has led us to explore the possibility of utilizing the technique to enhance transfection efficiency. We have identified electroporation parameters that improve the efficiency of DNA transfection in cutaneous wounds, and we have shown that electroporation itself does not impair wound healing. We are now on the threshold of exploring whether electroporation assisted transfection with DNA plasmid expression vectors for growth factors will be an effective modality for enhancing cutaneous wound healing. PMID- 15815833 TI - Stone soup as concocted by Professor Norman Rich. PMID- 15815834 TI - Popliteal vascular entrapment. PMID- 15815835 TI - In situ saphenous vein bypass--forty years later. AB - In situ saphenous vein bypass, which was performed first by Rob in 1959, but introduced by Hall in 1962, has been widely applied as an alternative to the reversed bypass. Now, forty years later, it seems appropriate to review its current place and technique. Of the presumed original benefits of the in situ operation, it is now clear that the hemodynamic flow of converging (in situ) versus diverging vein (reversed) boundaries plus better vein/artery size match are the main advantages. It is now popular belief that the reversed saphenous vein graft to the popliteal artery has no significant hemodynamic disadvantage because the vein is of comparable diameter at the knee and in the groin. In contrast, vein bypass from the groin to the ankle strongly favors the in situ procedure because of the convergence of the walls of the vein below the knee and better vein/artery size match. The most controversial facet of the in situ operation has been the question of valvulotome is superior to lyse valves and whether to perform the operation open as originally described, or closed to avoid skin complications from a long groin to ankle incision. Preoperative vein mapping is advantageous for all saphenous vein conduit operations. For open leg in situ procedures, skin bridges and incisions made directly over the vein, directed by mapping, will minimize skin complications. Use of endoscopic in situ techniques is ideal but only with an experienced endoscopist. Valve lysis has improved but is still not foolproof. PMID- 15815836 TI - Civil War vascular injuries. AB - As the result of the insistence of the Surgeon General during the United States Civil War, there was extensive documentation of injuries to major blood vessels and their resulting complications. The specific treatment of vascular injuries during the Civil War was ligation of the injured vessel or amputation. This was before there was any knowledge of the cause and prevention of infection. Overall, the results were dismal, with a mortality rate of nearly 60% for the more than 1000 soldiers treated by arterial ligation. The most important contribution of these medical reports was to define how the injuries should be diagnosed and managed. Many of the principles that developed as the result of this post-war review are as valid today as they were then. Unfortunately, it seems that many of these lessons have had to be relearned by the surgeons who have participated in each of our subsequent military conflicts. PMID- 15815838 TI - World War II in Alaska. PMID- 15815837 TI - The challenge of military surgical education. AB - Now that field hospitals and rescue stations have been provided with state-of-the art equipment, it is important to tailor the medical qualifications of military surgeons to the specific requirements of missions outside Germany. The objective of this article is to provide guidelines for a new training model. Einsatzchirurgie is defined as surgical treatment provided under restricted conditions in an unfamiliar environment. Its purpose is, first and foremost, to provide emergency treatment. The spectrum of Einsatzchirurgie, however, also encompasses maximum medical treatment on a case-by-case basis and emergency surgical treatment in a mass casualty situation. Training, for example, may consist of a 6-year basic training course in surgery followed by specialist training in abdominal surgery, traumatology, and courses in further disciplines- e.g., urology, gynecology, or neurosurgery. In addition to the qualifications required by the Landesarztekammem (professional organizations of German physicians at the federal countries level), military surgeons who are to become senior medical officers should also be qualified to provide immediate non surgical emergency care. In these times, the education of military surgeons remains a great challenge. Motivation to work as a surgeon in the armed forces with multiple deployments during the career requires enthusiasm and professionalism. The attractiveness of the training institution and the home hospital is key for successful work as a military surgeon. PMID- 15815839 TI - Combat casualty care research: from bench to the battlefield. AB - Hemorrhagic shock is the leading cause of death in civilian and combat trauma. Effective hemorrhage control and better resuscitation strategies have the potential of saving lives. The Trauma Readiness and Research Institute for Surgery (TRRI-Surg) was established to address the core mission of the Uniformed Services University, "Learning to Care for Those in Harm's Way," by conducting research to improve the outcome of combat casualties. This article highlights the salient achievements of this research effort in the areas of hemorrhage control, resuscitation, design and testing of devices, and some novel concepts such as the use of profound hypothermia. The impact of these basic science research findings on changes in military medical care and outcome of injured soldiers is also described. PMID- 15815840 TI - Airway stenting. AB - Interventional pulmonology, otherwise known as "airway stenting," has developed in the field of pulmonary medicine focused on using advanced bronchoscopic techniques to treat airway disorders. Tracheobronchial disorders can be caused by malignant or benign tumors, extrinsic compression, postintubation tracheal injuries, tracheobronchomalacia, or sequelae after tracheostomy. Tracheobronchial prostheses, known as airway stents, are used to palliate the effects of large airway obstruction. Specially designed stents are being used increasingly, not only in the airways, but also in the biliary tree, esophagus, urinary tract, and vascular system. There are two main types of airway stents currently available; tube stents made of silicone, and expandable metallic stents. Silicone stents are usually placed with the aid of a rigid bronchoscope while the patient is under general anesthesia. Unlike silicone stents, metal stents can be placed with a flexible bronchoscope. We examine the advantages and disadvantages of currently available stents and present our thoughts on the future development of airway stenting. PMID- 15815841 TI - Ischemic changes in evoked spinal cord potentials during profound hypothermic circulatory arrest in thoracic aortic surgery. AB - PURPOSE: We examined the changes in evoked spinal cord potentials (ESCP) during profound hypothermic circulatory arrest to estimate the safe ischemic time. METHODS: We monitored ESCPs during surgery for descending thoracic or thoracoabdominal aneurysms in five patients. Evoked spinal cord potential recordings were obtained before cooling (baseline), then every few minutes during circulatory arrest, and at the end of the operation. RESULTS: After circulatory arrest, the amplitude of ESCPs decreased with time. We calculated the simple linear regression between the amplitude of ESCPs and the circulatory arrest time by the least-squares method, and found a highly linear relationship between amplitude and arrest time in all five patients. The time until disappearance of ESCPs was estimated as 50.7 +/- 20.4 min (95% level of confidence). CONCLUSION: When an ESCP disappeared, ischemic spinal cord injury had occurred. This demonstrates the potential value of estimating the time of disappearance of ESCPs to prevent ischemic spinal cord injury during descending thoracic aortic surgery. PMID- 15815842 TI - Surgical options for malignant left-sided colonic obstruction. AB - PURPOSE: We evaluated the clinical results of different techniques of resection for malignant left-sided colonic obstruction. METHODS: The subjects of this prospective nonrandomized study were 63 consecutive patients who underwent surgery between 1995 and 2000 at a single institution. Patients with nonprimary colonic tumors, lesions located proximally to splenic flexure, peritonitis, perforation, or cecal necrosis were excluded. RESULTS: Segmental colectomy with primary anastomosis (CPA) was performed in 35 patients; with intraoperative colonic irrigation (ICI) in 19, and without ICI in 16. Total or subtotal colectomy (TSC) was performed in 8, and Hartmann's procedure (HP) was performed in 20. There were no differences in age, sex, comorbidity, time of symptoms, preoperative hospital stay, or tumor staging among the groups of patients defined by the different surgical techniques. The overall incidence of postoperative complications was 43%; postoperative mortality, 5%; anastomotic dehiscence, 12%; urgent reoperations, 12%; and readmissions, 5%, without significant differences among the treatment groups. However, the postoperative and total hospital stay were significantly shorter after ICI (P = 0.016 and P = 0.012, respectively). The overall 5-year survival was 42.7%. CONCLUSIONS: We think that segmental colectomy with anastomosis after intraoperative colonic irrigation is the most effective operative treatment for neoplastic left-sided colonic obstructions, considering its safety and cost-effectiveness. PMID- 15815843 TI - Repeat hepatectomy for recurrent colorectal metastases. AB - PURPOSE: To determine the risks and benefits of repeat hepatectomy for hepatic metastases from colorectal cancer. METHODS: During a recent 10-year-period, 106 patients underwent hepatectomy for hepatic metastases from colorectal cancer, in our hospital. Recurrence developed in the liver in 57 of these patients, 27 of whom underwent repeat hepatectomy. We reviewed the outcomes of these 27 patients. RESULTS: There were three complications after the first hepatectomy and six complications after the second hepatectomy, but there was no perioperative mortality after the first or second hepatectomy. The median survival from the date of second hepatectomy was 41 months with an actuarial 5-year survival rate of 48.7%. Patients who underwent repeat hepatectomy had significantly higher survival rates from the time of first hepatectomy than those who did not. Univariate analysis showed that among the prognostic factors of repeat hepatectomy, only a disease-free interval (DFI) between the first and second hepatectomy of more than 1 year was significantly predictive of a better outcome (P = 0.047). CONCLUSION: Repeat hepatectomy for recurrent colorectal metastases can be performed safely with acceptable mortality and morbidity rates, and can help to extend survival, if the DFI between the first and second hepatectomy is longer than 1 year. PMID- 15815844 TI - Granulocyte macrophage-colony stimulating factor improves impaired anastomotic wound healing in rats treated with intraperitoneal mitomycin-C. AB - PURPOSE: Intraperitoneal chemotherapy (IPCT) delivers higher local concentrations of cytotoxic drugs than intravenous (i.v.) chemotherapy, but it can adversely affect the healing of intestinal anastomoses if given in the early postoperative period. Intestinal anastomotic leakage is a serious surgical complication. Experimental and clinical studies have shown that the local administration of granulocyte macrophage-colony stimulating factor (GM-CSF) improves would healing. Therefore, we evaluated the effects of locally applied GM-CSF on anastomotic wound healing in rats treated with intraperitoneal mitomycin-C immediately after surgery. METHODS: We performed colon anastomoses in albino rats, which were then divided into three treatment groups. Group A was a control group that received no treatment, Group B was given intraperitoneal mitomycin-C postoperatively, and Group C was given intraperitoneal mitomycin-C with a local injection of GM-CSF postoperatively. We measured bursting pressures and hydroxyproline content, and histologically examined the resected anastomoses on postoperative day (POD) 3. RESULTS: Anastomotic healing was impaired after intraperitoneal mitomycin-C, but this was overcome by the injection of GM-CSF into the perianastomotic area. CONCLUSION: Local GM-CSF administration counteracts the detrimental effects of intraperitoneal mitomycin-C treatment on intestinal anastomoses in rats. PMID- 15815845 TI - Effect of nutritional state on postoperative axillary leakage after axillary lymph node dissection in a new rat model. AB - PURPOSE: Axillary leakage after axillary lymph node dissection (ALD) in breast cancer surgery is a frequent and troublesome complication. We conducted this study to establish an experimental model of axillary leakage after ALD; to evaluate axillary leakage in different nutritional states; and to examine the effects of dietary management on axillary leakage. METHODS: Twenty-eight Wistar rats were divided into two groups according to whether they were fed a high-fat diet or a low-fat diet for 14 weeks before ALD. After ALD, a suction drain was placed in the axilla and continuous suction was started. Axillary leakage was monitored for 9 days, and the drainage fluid and serum were analyzed biochemically. RESULTS: The body weights of the two groups were not significantly different before or after ALD. The amount of cumulative drainage fluid was significantly higher on postoperative days 7 and 9 in the high-fat group. The serum phosphate lipid, total cholesterol, and free cholesterol levels, and the free cholesterol level in the drainage fluid were also significantly higher in the high-fat group. CONCLUSIONS: This model is useful for reproducing lymphatic leakage after ALD. Our findings also suggest that a low-fat diet may be advantageous for patients undergoing ALD as part of breast cancer surgery. PMID- 15815846 TI - Unusual metastatic spread of follicular thyroid carcinoma: report of a case. AB - Concurrent skull and liver metastases from follicular thyroid carcinoma is a very rare event. We herein present the case of a 72-year-old woman who initially presented with a swelling in the right supraorbital region that proved to be metastasis from a well-differentiated follicular thyroid carcinoma of clear-cell type. The metastatic workup disclosed a huge liver metastasis and an additional metastasis in the left iliac fossa. The treatment of this patient included a total thyroidectomy, an excision of the skull lesion, and the administration of radioiodine therapy, as well as thyroid-stimulating hormone (TSH) suppression therapy. However, the course of her disease was relentless. Although well differentiated thyroid carcinoma tends to show an excellent course, the presence of metastatic disease leads to a very dismal prognosis. PMID- 15815847 TI - Sclerosing papillary carcinoma arising in a lingual thyroid: report of a case. AB - Ectopic lingual thyroid tissue is an uncommon congenital anomaly. Tumors with identical pathological characteristics to those arising in thyroid tissue may be present in ectopic locations, but there are very few cases of malignant ectopic thyroid tumors reported in the literature. We present a review of this phenomenon and report a case of papillary carcinoma of the base of the tongue, located in ectopic lingual thyroid tissue, in a 30-year-old woman. The patient's presenting symptoms were dysphagia and oral bleeding, and we performed radical resection of the neoformation at the base of the tongue with part of the muscles of the floor of the mouth and the body of the hyoid bone, as well as total thyroidectomy. Histological examination revealed a "sclerosing" papillary carcinoma. The patient was treated with 131I and substitutive thyroid hormonal therapy. An ultrasonogram done 5 years later showed bilateral laterocervical lymph node recurrence, which was effectively treated with bilateral laterocervical lymphectomy. PMID- 15815849 TI - Dedifferentiated liposarcoma of the pleura: report of a case. AB - We report a case of dedifferentiated liposarcoma of the pleura in a 59-year-old man who presented with pain in the right chest wall, 3 years after a routine chest X-ray showed a large tumorous mass. The resected tumor was composed of three distinct histological features: well-differentiated liposarcoma with low grade leiomyosarcomatous and low-grade osteosarcomatous components. To our knowledge, this is the first report of dedifferentiated liposarcoma of the pleura. PMID- 15815848 TI - Lung metastasis of thymoma manifesting as myasthenia gravis 12 years after thymomectomy: report of a case. AB - A 51-year-old woman, who had undergone thymomectomy for asymptomatic noninvasive thymoma 12 years before, was admitted to our hospital with blepharoptosis caused by myasthenia gravis. A chest computed tomogram (CT) showed an abnormal shadow in the right lower lung field and CT-guided needle biopsy revealed findings of a thymoma. We performed extended thymectomy and partial resection of the right lung using three different approaches, via a cervical incision and bilateral video assisted thoracoscopic surgery. The resected specimen was a lung metastasis of thymoma, and the residual mediastinal tissue showed no sign of malignancy. Because thymoma and post-thymomectomy myasthenia gravis can both recur, we recommend performing extended thymectomy or thymothymectomy, even for patients who are asymptomatic. PMID- 15815850 TI - Endovascular stent grafting for thoracic aneurysms in Jehovah's Witnesses: report of three cases. AB - There are few published reports on endovascular stent grafting for thoracic aneurysms in Jehovah's Witnesses. Between 2001 and 2003, we performed endovascular stent grafting for a thoracic aneurysm in three patients of the Jehovah's Witness faith. Two patients had a thoracic aortic aneurysm and one had a chronic type-B dissection. The stent graft was constructed from a self expanding Z-stent and thin-walled woven polyester fabric. None of the patients required perioperative blood transfusion, there was no postoperative endoleak, and all recovered uneventfully and were discharged from hospital. Thus, stent graft repair of thoracic aneurysms in Jehovah's Witnesses is feasible and can be achieved without the need for blood transfusion. PMID- 15815851 TI - Abdominal compartment syndrome causing respiratory failure during surgery for a ruptured descending thoracic aneurysm: report of a case. AB - Elevated intra-abdominal pressure causing widespread organ dysfunction is known as abdominal compartment syndrome (ACS). The subject of our case report is a 64 year-old man who underwent repair of a ruptured descending thoracic aortic aneurysm (TAA) under deep hypothermic circulatory arrest. During the operation, decompression laparotomy was required to relieve intra-abdominal hypertension causing respiratory failure, before the patient could be weaned off cardiopulmonary bypass. We report this case to alert surgeons to the fact that ACS can occur during surgery on the thoracic aorta, especially if massive fluid resuscitation is required and venous drainage for extracorporeal circulation is less than optimal. Early recognition and prompt decompression by laparotomy is essential to save the life of the patient. PMID- 15815852 TI - Primary malignant fibrous histiocytoma of the ascending colon: report of a case. AB - We report a rare case of primary malignant fibrous histiocytoma (MFH) of the ascending colon. A 66-year-old man presented to our hospital with epigastralgia, and abdominal ultrasonography and computed tomography showed a large soft-tissue mass in the ascending colon. Barium enema and endoscopic examination showed a huge tumor in the ascending colon. At laparotomy, we found a tumor in the ascending colon and performed a right hemicolectomy with en bloc lymph node dissection. The resected specimen contained a tumor measuring 14.5 x 8.0 x 4.5 cm, the cut surface of which was yellowish. Based on histological and immunohistological studies, the tumor was diagnosed as MFH of the ascending colon. To our knowledge, only 20 cases of colorectal MFH, including our case, have been documented, which we review following this case report. PMID- 15815853 TI - Primary squamous cell carcinoma of the liver arising from a complex liver cyst: report of a case. AB - A 65-year-old man who had received radiation therapy for nasopharyngeal cancer (NPC) 3 years earlier presented with a 3-week history of right upper quadrant abdominal pain and a feeling of fullness. There had been no evidence of metastasis on his follow-up examinations. Computed tomography scan showed a huge complex cyst with septa in the right hepatic lobe, and we performed an extended right hepatectomy to relieve his symptoms. Pathological examination revealed a large hepatic cyst with malignant cells along the cyst wall. The cytokeratin stain and CK-14 stains were positive, indicating an undifferentiated squamous cell carcinoma (SCC). The final diagnosis of primary SCC of the liver was confirmed by the clinical pathological features and negative in situ hybridization of Epstein-Barr ribonucleic acids (EBERs). We used EBERs to determine whether the cystic tumor was a primary lesion or a metastatic lesion from the previous NPC. PMID- 15815854 TI - Pseudoaneurysm of the dorsal pancreatic artery with obstruction of the celiac axis after pancreatoduodenectomy: report of a case. AB - Pancreatoduodenectomy can be difficult in patients with obstruction of the celiac axis because of the development of collateral arteries around the head of the pancreas. Bleeding from a pseudoaneurysm is a rare and serious complication of pancreatoduodenectomy. We report a case of a pseudoaneurysm of an enlarged dorsal pancreatic artery, which formed as a result of obstruction of the celiac axis after pancreatoduodenectomy. We assumed that a weakness in the wall of this dorsal pancreatic artery caused by the dissection led to the formation of the aneurysm. Although transcatheter arterial embolization (TAE) could not treat the aneurysm or stop bleeding from the aneurysm completely, it achieved transient hemostasis, whereby the patient's condition improved, making laparotomy safe. This case demonstrates that the dissection of arteries that have developed around the head of the pancreas must be considered in patients with obstruction of the celiac axis. PMID- 15815855 TI - Clinicopathological and immunohistochemical features of extragastrointestinal stromal tumors: report of two cases. AB - CD117 (c-kit proto-oncogene protein product) is expressed in most gastrointestinal stromal tumors (GISTs) and plays a crucial role in the pathogenesis and treatment of this disease. However, the clinicopathological and immunohistochemical features of CD117-positive mesenchymal tumors without connection to the gastrointestinal tract, known as extragastrointestinal stromal tumors (EGISTs), are not well documented because these tumors are rare. We describe the clinicopathological and immunohistochemical features of two cases of EGIST and compare them with those of GIST. Of the 1855 abdominal or esophageal tumors resected during the past 10 years at our hospital, 23 were GISTs and 2 were EGISTs. The clinicopathological or immunohistochemical characteristics do not seem to differ remarkably between EGISTs and GISTs. Although rare, CD117 positivity should be tested in abdominal mesenchymal tumors that have no connection to the gastrointestinal tract. The clinicopathological features of CD117-positive abdominal mesenchymal tumors may not depend on whether the tumor is connected to the gastrointestinal tract. PMID- 15815856 TI - Esophageal delta-shaped anastomosis: a new method of stapled anastomosis for the cervical esophagus and digestive tract. AB - We describe a new technique of anastomosis between the esophagus and digestive tract, developed as an extension of a functional end-to-end anastomosis, and named the "esophageal delta-shaped anastomosis." We also report its short-term clinical results. We used this anastomotic technique in nine patients undergoing surgery for thoracic esophageal carcinoma. The reconstruction route was posterior mediastinal (n = 5), antethoracic (n = 3), or retrosternal (n = 1), and the organs for esophageal replacement were the gastric tube (n = 7) or the colon (n = 2). When using the gastric tube, a linear stapler is inserted into two intestinal windows; one created at the stump of the esophagus and the other created in the posterior wall near the greater curvature of the gastric tube. The stapler is fired, ensuring that the staple line does not sharply cross the staple line of the lesser curvature of the gastric tube. Two linear staplers are then used to close the window transversally to form a delta shape. There were no complications related to the anastomosis. The esophageal delta-shaped anastomosis is a simple and safe method for esophagogastrostomy or esophagoenterostomy, which uses a combination of introverted and extroverted stapling, resulting in a larger lumen not limited by the lumen of the esophagus or the organs used for esophageal replacement. PMID- 15815857 TI - Age-related quadriceps-dominant muscle atrophy and incident radiographic knee osteoarthritis. AB - Muscle atrophy is common in the elderly. However, the etiologic role of muscle atrophy associated with osteoarthritis of the knee has not been studied in detail. We assessed the association between age-related muscle atrophy around the knee joint and incident radiographic osteoarthritis of the knee. Twenty-one women in their thirties and 17 women in their sixties participated. They had no history, symptoms, or objective findings of any knee problems, and none of the participants was limited in performing daily activities. Radiographs of the knee joint were graded for the presence of osteoarthritis, and the cross-sectional imaging around the knee joint was carried out using computed tomography. Incident radiographic osteoarthritis was observed in 6 of the 17 women in their sixties. Quadriceps-dominant muscle atrophy was marked in the elderly women with radiographic osteoarthritis. In a multivariate analysis, the risk of incident radiographic osteoarthritis of the knee was significantly increased among women with a higher hamstrings/quadriceps cross-sectional area ratio. The results suggest the possibility that age-related quadriceps-dominant muscle atrophy may play a role in the pathogenesis of osteoarthritis of the knee. PMID- 15815858 TI - Repair of cartilage defects and torn acetabular labrum in hip joints after conventional osteotomy: evaluation by follow-up arthroscopy. AB - The purpose of this study was to evaluate recovery from cartilage damage and labral lesions after osteotomy of hip joints. Conventional osteotomy for dysplastic hips was performed on 38 hips. Hip arthroscopy preceded the osteotomy, which in turn was followed up 18.9 months later on average by arthroscopy. We assessed the Harris hip score (HHS) and radiological and arthroscopic findings before and after osteotomy. The average HHS was improved from 64.9 to 88.5. Cartilage repair was observed on the acetabular side of five hips (13.2%) and on the femoral side of four (10.5%). At follow-up arthroscopy, the acetabular labrum had adhered to the capsule in 11 hips. Acetabular adhesion may increase stability of the joint. We conclude that the effects of conventional osteotomy are limited, and additional surgery might be needed in the future. PMID- 15815859 TI - Insertional torque of the lumbar pedicle screw during surgery. AB - The purposes of this study were to determine if the intraoperative insertional torque of pedicle screws correlates with the degree of osteoporosis and if insertional torque can be used as a predictor of screw loosening and clinical results in elderly patients. Pedicle screw fixation was performed in 25 patients. Their mean age at the time of surgery was 72.2 years (range 65-79 years). The mean follow-up period was 2.8 years (range 1.0-4.8 years). The insertional torque of the pedicle screws was measured using a Kannon-type torque wrench with a special connector. In the present study, 136 screws were evaluated. The mean insertional torque was 12.2 +/- 5.0 kgf x cm. A negative relation was found between insertional torque and the grade of osteoporosis. Roentgenograms showed the radiolucency of 18 screws in eight patients. Radiolucency appeared as early as 6 months postoperatively. However, there was no significant relation between insertional torque and the presence of screw loosening. Furthermore, there was no significant difference in the clinical results between patients with low insertional torque and those with high insertional torque. This study demonstrated that intraoperative insertional torque is not an objective predictor of screw loosening or clinical results. PMID- 15815860 TI - Intraoperative insertion torque of lumbar pedicle screw and postoperative radiographic evaluation: short-term observation. AB - The correlation between the insertion torque of a lumbar pedicle screw and the mechanical stability of the screw in the bone has been mentioned in in vitro studies. The purpose of this study was to confirm the factors affecting the insertion torque of such screws in vivo. Also, the contribution of insertion torque to the initial stability of the fusion area was to be analyzed in vivo. A series of 23 cases representing 50 lumbar vertebrae were included in this study, in which we examined bone mineral density using quantitative computed tomography (CT) prior to operation. Two screw shapes were utilized, with the insertion torque for each screw measured at two points in time. The correlation between insertion torque and mineral density was investigated. Screw positions were confirmed on postoperative CT scans, and the effect of the screw thread cutting into the cortex bone was investigated. Radiographic changes at three points during a period of 3 months were also measured, and we then evaluated the interrelations between these changes and insertion torque. Furthermore, the relation between insertion torque and instability at 3 months was investigated. Correlations of insertion torque and bone mineral density depended on screw shape. There was no correlation found with mineral density in the case of cylindrical screws. Insertion torque was not affected by the screw thread cutting into the cortex of bone. As for postoperative alignment changes, no definitive trends could be ascertained, and no interrelations with torque and alignment changes were observed. There is a possibility that insertion torque was related to early-stage stability, but no statistical relation could be determined. PMID- 15815861 TI - A computer simulation of rotational acetabular osteotomy for dysplastic hip joint: does the optimal transposition of the acetabular fragment exist? AB - For young patients who have early signs of coxarthrosis resulting from acetabular dysplasia, periacetabular osteotomies for correcting abnormal stress distribution can be useful for preventing the progression of the disease. However, it is difficult to confirm the optimal transposition of the osteotomized acetabular fragment. To deal with this problem, we devised a computer program to support preoperative planning. Hip images obtained by computed tomography were loaded into our program, and a three-dimensional voxel model was created. Then, osteotomy was simulated and the pressure distribution was analyzed with a rigid body spring analysis (computational nonlinear mechanical analysis). The three dimensional pressure distributions in seven dysplastic hips were evaluated before and after virtual rotational acetabular osteotomy. A peak pressure was calculated for every 5 degrees of rotation of the acetabular fragment. The peak pressure decreased gradually and increased again afterward, indicating the optimal transposition of the acetabular fragment. The postoperative peak pressure decreased to about 40% in the most improved case. This program allows the hip joint mechanics to be evaluated easily so that the advantages and disadvantages of various surgical methods can be examined biomechanically prior to surgery. PMID- 15815862 TI - Spondylolisthesis with postural slip reduction shows different motion patterns with video-fluoroscopic analysis. AB - To compare the motion of lumbar spondylolisthesis between the standing and decubitus positions and to identify the radiological parameters concerning segmental instability, the lumbar sagittal motion of 5 volunteers and 14 patients was recorded using video-fluoroscopy in both positions. Still images at three frames per second were captured, and the motion pattern was compared between the two positions. Patients were classified into two subgroups according to the degree of postural slip reduction. In the volunteer group, the motion patterns were not particularly different between the two positions. In the small postural reduction group, the motion patterns were not particularly different between the two positions, except in one patient for whom the range of motion (ROM) at the upper adjacent level increased in the decubitus position. In the large postural reduction group, a motion pattern change was observed. The ROM at the slip level in three patients increased, and in the remaining four patients the angle at the midpoint in ROM at the slip level shifted to the extension side in the decubitus position. This condition (anterior slip being reduced in the supine position, with disc angle and average disc height increased) is thought to result from anterior column deficiency and the vertical laxity of the surrounding soft tissues. A biomechanically stable fusion technique is recommended for surgical management. PMID- 15815863 TI - Comparative evaluation of two two-dimensional gel electrophoresis image analysis software applications using synovial fluids from patients with joint disease. AB - The proteomic composition of synovial fluid (SF) may hold clues to understanding the molecular basis of arthritis. However, the highly viscous nature and proteomic complexity of SF present a challenge when analyzing results obtained by two-dimensional gel electrophoresis (2D-GE). Several software applications are available for analyzing 2D-GE images. Despite inherent strengths and weaknesses, no comparison between these applications has been reported using SF or any human fluid specimens. We evaluated two common software packages--PDQuest and Progenesis Workstation--for spot detection, matching, and quantitation of 2D-GE images of SF from four patients with arthritic disease. Initially, whole 2D-gel images were analyzed for spot detection, which suggested that PDQuest is more consistent than Progenesis; however, PDQuest appeared to require more user intervention than Progenesis. Subsequently, two small areas (spots well resolved and spots not well resolved) were selected from each gel image, which were analyzed by the software for spot detection, matching, volume, and resolution. These analyses suggest that both tools can quantify well-resolved spots relatively consistently when compared with manual spot detection (the "gold standard"). The "3D viewer" option offered by both tools enables correct spot identification and matching. The strengths and weaknesses of these computer tools can provide guidance in the choice of a particular workstation for identifying biomarkers of arthritis. PMID- 15815865 TI - Displaced femoral neck fracture in children and adolescents: closed versus open reduction--a preliminary study. AB - Twenty-six children and adolescents with displaced femoral neck fractures who were treated surgically from 1992 to 2002 were included in this study. Among them, 14 were male and 12 were female. Altogether, 10 of them had a transcervical type of fracture, and the other 16 had a cervicotrochanteric type of fracture. The average age of the patients was 10.8 years (3-17 years). Seventeen were treated by closed reduction and internal fixation (group A), and nine required open reduction and internal fixation (group B). In group A, there were good results in 11 cases (65%), a fair result in one case (6%), and poor results in 5 cases (29%). In group B, there were good results in seven cases (78%) and fair results in the other two cases (22%). The average fracture union times in groups A and B were 12.6 and 10.2 weeks, respectively. Thus, there was a favorable outcome in group B. However, statistical analysis of data in this preliminary study did not reveal any significant difference in the surgical outcome between the two groups. PMID- 15815864 TI - Pelvic motion during total hip arthroplasty with translateral and posterolateral approaches. AB - We devised a special instrument to assess intraoperative pelvic motion and used this device to measure intraoperative pelvic motion in three dimensions. A total of 100 total hip arthroplasties (THAs) were performed using this device. Two approaches were utilized: 52 THAs were performed with the posterolateral approach and the remaining 48 with the translateral approach. The average angle of pelvic tilt in the THAs with the translateral approach was less than that with the posterolateral approach. The average internal rotation angle was 1.75 degrees with the translateral approach and 14.25 degrees with the posterolateral approach. With the posterolateral approach, the internal rotation of the pelvis frequently occurred during retraction of the femur using a Hohmann's retractor placed at the anterior rim of the acetablum, with flexion and internal rotation of the affected hip. PMID- 15815866 TI - Treatment of proximal humeral fractures with a new intramedullary nail. AB - We have developed a new intramedullary nail (All-in-One Nail) that is specifically designed for elastic fixation of a surgical neck fracture of the humerus. This nail has three pins and a plate, and the pins become divergent as the nail is inserted into the marrow cavity through the deltoid tuberosity and then advanced to the humeral head. We reviewed 31 patients with an age range of 58-91 years (mean 75 years) who underwent surgery with this nail for A3.2 (two part), B2.2 (three-part), or C2.1 (valgus-impacted) fractures of the proximal humerus. Bony union was obtained in 29 patients, and these patients had a Japanese Orthopaedic Association shoulder score of 60-92 (mean 78) in a follow-up examination performed 6-32 months (mean 13 months) postoperatively. The All-in One Nail system is easy to use, provides an acceptable clinical outcome, and is a treatment option for unimpacted surgical neck fractures and valgus-impacted fractures of the humerus. PMID- 15815867 TI - Treatment of femoral shaft fractures in children using intramedullary Kirschner wire pinning. AB - From 1996 to 2002 we used intramedullary Kirschner wires to treat 19 femoral shaft fractures in 19 children (11 boys, 8 girls). Their ages ranged from 2 to 13 years (mean 5.8 years). The mean follow-up time was 3 years (range 4 months to 4 years 9 months). The technique was intramedullary pinning using a 2.5- to 3.0-mm Kirschner wire introduced through the greater trochanter followed by spica casting. All cases achieved bone union within 3 months. The mean angular deformity was 1.7 degrees (range 0 degrees -5 degrees ), and one patient had an external rotational deformity. The mean overgrowth was 7.5 mm, but no patient had residual problems during activities of daily living. Our method has the following advantages: no need of traction, leaving bed earlier, ease of nursing, fewer radiographic examinations, less angular deformity. It also has several disadvantages: the need for an operation under general anesthesia, the need for metal removal, and an operation scar. This method can be considered as one option for treating femoral shaft fractures in children. PMID- 15815868 TI - Comparison of bioengineered human bone construct from four sources of osteogenic cells. AB - Osteoprogenitor cells have been reported to be present in periosteum, cancellous and cortical bone, and bone marrow; but no attempt to identify the best cell source for bone tissue engineering has yet been reported. In this study, we aimed to investigate the growth and differentiation pattern of cells derived from these four sources in terms of cell doubling time and expression of osteoblast-specific markers in both monolayer cells and three-dimensional cell constructs in vitro. In parallel, human plasma derived-fibrin was evaluated for use as biomaterial when forming three-dimensional bone constructs. Our findings showed osteoprogenitor cells derived from periosteum to be most proliferative followed by cortical bone, cancellous bone, and then bone marrow aspirate. Bone-forming activity was observed in constructs formed with cells derived from periosteum, whereas calcium deposition was seen throughout the constructs formed with cells derived from cancellous and cortical bones. Although no mineralization activity was seen in constructs formed with osteoprogenitor cells derived from bone marrow, well-organized lacunae as would appear in the early phase of bone reconstruction were noted. Scanning electron microscopy evaluation showed cell proliferation throughout the fibrin matrix, suggesting the possible application of human fibrin as the bioengineered tissue scaffold at non-load-bearing sites. PMID- 15815869 TI - Corrosion of spinal implants retrieved from patients with scoliosis. AB - Spinal implants retrieved from 11 patients with scoliosis were examined. All the implants were posterior instrumentation systems made of 316L stainless steel and composed of rods, hooks, and crosslink connectors. Corrosion was classified into grades 0 to 3 based on macroscopic findings of the rod surface at the junction of each hook or crosslink connector. Grade 0 was defined as no sign of corrosion, grade 1 as surface discoloration, grade 2 as superficial metal loss, and grade 3 as severe metal loss. The depths and characteristics of metal loss areas were examined. Spinal implants showed more corrosion after long-term implantation than after short-term implantation. Corrosion was seen on many of the rod junctions (66.2%) after long-term implantation, but there was no difference between the junction at the hook and those at the crosslink connector. It is thought that intergranular corrosion and fretting contributed to the corrosion of implants. The current study demonstrated that corrosion takes place at many of the rod junctions in long-term implantation. We recommend removal of the spinal implants after solid bony union. PMID- 15815870 TI - Study of blood metabolism and urinary excretion of chymopapain following intradiscal injection using a high-sensitivity enzyme immunoassay. AB - To develop chymopapain-induced chemonucleolysis as an established treatment, it is necessary to determine the kinetics of chymopapain in blood and urine following intradiscal injection. To investigate the rate of blood metabolism and urinary excretion of chymopapain following intradiscal injection, we developed a high-sensitivity enzyme immunoassay for chymopapain. The sensitivity for this assay was 1 pg/tube (40 amol). After injecting chymopapain into the nucleus pulposus of humans, levels of blood chymopapain were measured by enzyme immunoassay. The level of chymopapain in blood decreased gradually, with a half life of 2-3 days. The half-life for urinary excretion was a little longer, at 3 days. It was also found that chymopapain in blood was not present as a free molecule but formed a complex that had a molecular weight of about 120 kDa. These findings suggest that most chymopapain would not have activity in blood. PMID- 15815871 TI - Distribution of sodium channels during nerve elongation in rat peripheral nerve. AB - A number of studies have investigated electrophysiological and morphological changes of peripheral nerves during gradual elongation. There has been, however, no report on the distribution of sodium channels at Ranvier's nodes during peripheral nerve elongation. We investigated peripheral nerve injury after the gradual elongation of rat sciatic nerves. Indirect nerve elongation was induced by leg lengthening at a rate of 3 mm/day by 15 or 30 mm. At 7 days after the leg lengthening, the electrophysiological properties of sciatic nerves, the ultrastructures of the Ranvier's nodes and axons, and the distribution of voltage dependent sodium channels were examined. In the control nerves, most sodium channels were localized at Ranvier's nodes in myelinated axons, providing the physiological basis of saltatory conduction. In the elongated nerves, both the amplitude and conduction velocity of compound nerve action potential decreased following leg lengthening. The elongated nerves also showed paranodal demyelination in Ranvier's nodes longer than those in the control group. In addition, the distribution of sodium channels became diffuse or disappeared at Ranvier's nodes of elongated nerves. The diffuse distribution and/or disappearance of sodium channels may underlie the electrophysiological changes in compound nerve action potential induced by nerve elongation. PMID- 15815872 TI - Diabetes with hyperlipidemia: a risk factor for developing joint contractures secondary to immobility in rat knee joints? AB - Several studies have reported that diabetes may cause limited joint motion and often combines with hyperlipidemia (HL). This study clarifies that joint contracture can be produced by diabetes, HL, or both. The experimental animal model, a rat with diabetes and HL, was developed by streptozosin injection and addition of 1% cholesterol to the food. One hindlimb each from the diabetes group, the HL group, the combined diabetes/HL group, and the control group was immobilized for 2, 3, or 4 weeks in the Phase I study. Diabetes/HL rats were used 4 weeks after immobilization in a Phase II study. Joint motion, elasticity of the knee joint capsule, and contractile function of the gastrocnemius muscle were measured. Gastrocnemius muscle was observed histologically by H&E and ATPase staining. Limited joint mobility was observed 4 weeks after immobilization only in the diabetes/HL group. The intraarticular pressure from the saline injection was lower, and the contractile function of the gastrocnemius muscle decreased in this group. Atrophy of type II fiber was observed in the gastrocnemius muscle. This restriction of joint mobility may depend on skeletal muscle degeneration around the joint rather than on an intraarticular lesion. PMID- 15815873 TI - Simultaneous rupture of the quadriceps tendon and contralateral patellar tendon in a patient with chronic renal failure. AB - Simultaneous quadriceps and patellar tendon rupture is rare. Mechanical factors and coexisting systemic and local factors are taken into consideration in the pathogenesis of these ruptures. In patients with some chronic systemic diseases, simultaneous rupture can occur spontaneously or with minor traumas. We present a case of simultaneous quadriceps and patellar tendon rupture in a 21-year-old man with chronic renal failure in this report. He was treated surgically by osseotendinous repair with suture anchors and supplemental cerclage wire fixation on both sides. He regained his normal knee joint functions 18 months after the operation. PMID- 15815874 TI - High-grade L5 spondylolisthesis associated with dural ectasia in neurofibromatosis. AB - A 15-year-old girl was admitted to our hospital with severe low back pain. She had scoliosis dextra and tight hamstrings. A plain radiograph showed high-grade L5 spondylolisthesis with vertebral scalloping from the fourth lumbar to the first sacral vertebra. L5 wide laminectomy and posterior lumbar interbody fusion by iliac bone graft was performed using the Galveston method of sacral fixation and a pedicle screw system. The rod and hook system was used from T9 to the bilateral iliac wing. We added posterolateral fusion from T10 to S1 by autograft and allograft. The patient became pain free and was able to return to student life. Three years after surgery, the radiographs demonstrated good bony fusion; however, careful long-term follow-up is needed. PMID- 15815875 TI - Autologous platelets and articular surface repair in an experimental model. PMID- 15815877 TI - The use of the point of care Helena ICHOR/Plateletworks and the Accumetrics Ultegra RPFA for assessment of platelet function with GPIIB-IIIa antagonists. AB - OBJECTIVE: To evaluate a newly modified rapid platelet function analysis system (ICHOR/ Plateletworks) and to compare the results obtained with those of traditional light transmission aggregometry (LTA), and the Ultegra/RPFA system. BACKGROUND: Anti-platelet therapy is standard of care for patients as an adjunct to percutaneous coronary intervention (PCI) or for medical management of non-ST elevation acute coronary syndromes (NSTE ACS). Recent clinical trial results suggest that the three currently approved platelet GPIIb-IIIa receptor antagonists, eptifibatide, tirofiban and abciximab, may vary in extent of inhibition of platelet aggregation (IPA) at the approved doses. Thus, pharmacodynamic evaluations of these agents to determine the extent of platelet function inhibition, especially during the periprocedural time of a cardiac intervention, are necessary. A rapid measurement method as a surrogate for LTA, the current gold standard, would be ideal in order to have the option for dose monitoring or adjustment prior to or during an intervention. The Helena ICHOR/ Plateletworks may be useful for point of care testing. METHODS: Blood samples collected in D-Phe-Pro-Arg-chloromethyl ketone dihydrochloride (PPACK) anticoagulant were treated with increasing concentrations of eptifibatide, tirofiban or abciximab. LTA was carried out in conjunction with the ICHOR/Plateletworks, using a modified method, and Accumetrics Ultegra with RPFA cartridges. RESULTS: This study demonstrated that platelet inhibition measured by the ICHOR/Plateletworks mirrored the level of IPA obtained with LTA. In contrast, the Ultegra system had less correlation when compared to LTA at inhibition levels < 90%. CONCLUSIONS: Based on these data, the ICHOR/ Plateletworks utilized under modified guidelines may serve as a surrogate for LTA when rapid measurements are necessary.A rapid platelet function measurement method as a surrogate for light transmission aggregometry (LTA), the current gold standard, is ideal in order to have the option for GPIIb-IIIa antagonist dose monitoring or adjustment prior to or during a coronary intervention. A newly modified rapid platelet function analysis system (ICHOR/Plateletworks was evaluated and compared to the results obtained with traditional light transmission aggregometry (LTA), and the Ultegra/RPFA system. Blood samples collected in D-Phe-Pro-Arg-chloromethyl ketone dihydrochloride (PPACK) anticoagulant were treated with increasing concentrations of eptifibatide, tirofiban or abciximab. LTA was carried out in conjunction with the ICHOR/Plateletworks, using a modified method, and Accumetrics Ultegra with RPFA cartridges. Based on these data, the ICHOR/Plateletworks utilized under modified guidelines may serve as a surrogate for LTA when rapid measurements are necessary. PMID- 15815878 TI - Antibodies to platelet factor 4/heparin are associated with elevated endothelial cell activation markers in patients with acute coronary ischemic syndromes. AB - BACKGROUND: We postulated that antibodies to platelet factor 4/heparin complex lead to a heightened inflammatory state, contributing to an increased risk of recurrent thrombotic events. METHODS: We analyzed serum from a subset of patients in the placebo/unfractionated heparin arm of the GUSTO IV-ACS trial who had prior heparin exposure. We selected 109 patients with the 30 day primary endpoint (death, MI or revascularization) and an equal number of controls, excluding patients with thrombocytopenia. Anti-platelet factor 4/heparin antibodies and inflammatory markers (sVCAM-1, sICAM-1, sE-selectin, sP-selectin, us-CRP, IL-6) were measured on serum samples. RESULTS: Patients with anti-PF4/heparin antibodies were more likely to have death or MI (30.4% vs. 11.3%, p = 0.01), or MI (21.7% vs. 6.2%, p = 0.01) than patients who were antibody negative. In a multiple logistic regression model that included inflammatory markers and clinical risk factors, antibody to PF4/heparin was a strong predictor of 30 day MI (odds ratio: 9.0; 95% confidence intervals 2.1-38.6; p < 0.01), with IL-6 being the only other predictor (odds ratio: 1.1; 95% confidence intervals 1.0 1.2; p = 0.03). Antibody positive patients had higher levels of sVCAM-1 (892 +/- 263 microg/l versus 780 +/- 228 microg/l; p = 0.04) and sICAM-1(246 +/- 50 microg/l versus 222 +/- 71microg/l; p = 0.02) than antibody-negative patients. CONCLUSIONS: Antibodies to the platelet factor 4/heparin complex were associated with elevated levels of endothelial but not platelet activation markers, or markers of a systemic inflammatory state. Anti-PF4/heparin antibodies were associated with a 9-fold increased risk of recurrent MI at 30 days. We measured soluble cell adhesion molecules, CRP and IL-6 from 218 non-thrombocytopenic patients, 23 of whom had antibodies to PF4/heparin. Antibody positive patients had higher levels of sVCAM-1 (892 +/- 263 microg/l versus 780 +/- 228 microg/l; p = 0.04) and sICAM-1(246 +/- 50 microg/l versus 222 +/- 71 microg/l; p = 0.02). In a multiple logistic regression model, antibody to PF4/heparin was a predictor of 30 day MI (odds ratio: 9.0; 95% CI: 2.1-38.6; p < 0.01). The presence of antibodies to PF4/heparin, even in the absence of thrombocytopenia, is a stronger predictor of 30 day MI than clinical variables or inflammatory markers. PMID- 15815879 TI - Losartan and simvastatin inhibit platelet activation in hypertensive patients. AB - BACKGROUND: Diabetic patients also show hypercoagulability and platelet hyperaggregability, with increased levels of platelet activation-markers such as P-selectin (CD62P) and platelet-derived microparticles. We investigated the effects of losartan and simvastatin on circulating levels of platelet activation markers, microparticles, soluble selectins, and soluble cell adhesion molecules in hypertensive and hyperlipidemic patients with or without Type 2 diabetes. METHODS: The subjects included 25 normotensive healthy controls and 41 hypertensive patients. The 41 hypertensive patients were divided into three groups: group A had hypertension and hyperlipidemia (n = 11), group B had hypertension and Type 2 diabetes (n = 14), and group C had hypertension, hyperlipidemia, and diabetes (n = 16). Losartan was administered to all of the patients at a dose of 50 mg/day for 24 weeks. In addition, simvastatin was administered to the hyperlipidemic patients at a dose of 10 mg/day for 24 weeks. RESULTS: There were significant differences in the levels of CD62P, CD63, PAC-1, platelet microparticles, endothelial microparticles, sE-selectin, and sVCAM-1 between the hypertensive patients and healthy controls. These markers were all significantly increased in hypertensive and hyperlipidemic patients with Type 2 diabetes. In hypertensive patients with diabetes, CD62P, CD63, PAC-1, platelet and endothelial microparticles, and soluble adhesion markers were all decreased by losartan monotherapy. The decrease of each marker in hypertensive and hyperlipidemic patients given combined therapy with losartan plus simvastatin was greater among those with than without Type 2 diabetes. Low-density lipoprotein was decreased significantly by simvastatin and was correlated with CD62P or platelet microparticles in all of the patients. CONCLUSION: Administration of losartan plus simvastatin to hypertensive and hyperlipidemic patients with Type 2 diabetes may prevent the development of cardiovascular complications caused by activated platelets and microparticles via another mechanism in addition to reduction of the blood pressure or lipid levels. PMID- 15815880 TI - Bleeding complications after contemporary pharmacoinvasive therapy for ST elevation myocardial infarction. AB - BACKGROUND: Pharmacoinvasive therapy for the treatment of ST elevation myocardial infarction (STEMI) is a strategy that combines early restoration of coronary flow via pharmacologically induced thrombolysis with subsequent, prompt percutaneous coronary intervention (PCI). Prior studies suggesting a heightened bleeding risk of PCI performed early after fibrinolysis predated contemporary pharmacoinvasive practice including use of femoral closure devices (CD), fibrin specific thrombolytics, lower doses of heparin and stents. METHODS: Consecutive patients were included in this retrospective registry study if they underwent emergent PCI for ST elevation myocardial infarction (STEMI) followed by immediate use of a groin closure device. Between Oct 1, 2002 and Jan 1, 2003, 27 patients were treated with immediate use of CD after post-thrombolytic PCI, performed within 12 hours of thrombolysis (pharmacoinvasive group). 58 patients were treated with immediate use of CD after primary PCI for STEMI. The two groups were compared with respect to the incidence of successful groin closure, bleeding complications, and clinical outcomes. Bleeding events were categorized according to the TIMI criteria. All baseline clinical and treatment variables were compared between the two groups to determine and the association of these variables (including use of thromblytic therapy) with TIMI major and TIMI minor bleeding was determined. RESULTS: Pharmacoinvasive recanalization with PCI occurred 348 +/ 183 minutes after initiation of fibrinolytic therapy. Glycoprotein IIb/IIIa inhibitors were used less frequently in the patients treated with a thrombolytic agent (59% vs. 90%, p < 0.01). Successful immediate hemostasis was obtained with CD in greater than 85% of patients in both groups (89% for pharmacoinvasive group vs. 86% for primary PCI group, p = 0.89). No patient required vascular surgical intervention. TIMI major bleeding and transfusion requirements were less than 5% in both groups. Antecedent thrombolytic therapy was not a predictor of bleeding complications after PCI. CONCLUSIONS: Use of CD as part of a contemporary pharmacoinvasive strategy is associated with a low rate of major bleeding complications. PMID- 15815881 TI - Vascular endothelial tissue factor pathway inhibitor kinetics in culture following exposure to DX-9065a--a selective and direct factor Xa inhibitor. AB - BACKGROUND: Tissue factor (TF), a membrane-bound glycoprotein that initiates blood coagulation by allosteric activation of factor (f) VII, is regulated predominantly by tissue factor pathway inhibitor (TFPI). Because vascular endothelial cells synthesize and constitutively secrete TFPI and fXa may directly influence its cellular clearance, we sought to determine the effects of DX-9065a, a direct and selective fXa inhibitor, on TFPI kinetics in culture. METHODS/RESULTS: Human umbilical vein endothelial cells were grown to confluence and incubated with unfractionated heparin (1.0 U/mL), enoxaparin (1.5 U/mL), or DX-9065a at low (10 ng/ml), moderate (30 ng/ml), or high (90 ng/ml) concentrations. Compared to control, increases in TFPI were seen with both unfractionated heparin (182% higher, p < 0.001) and enoxaparin (194% higher, p < 0.001). Low concentration DX-9065a did not increase TFPI levels above control (0.8% higher, p = 0.91). In contrast, moderate and high concentrations produced 124% higher (p < 0.001) and 198% higher (p < 0.001) TFPI concentrations than control, respectively. CONCLUSIONS: DX-9065a increases vascular endothelial cell TFPI concentrations in human tissue culture. Although the mechanism has yet to be established, decreased fXa availability may limit fXa-TFPI complex formation and its subsequent cellular uptake. Whether increased surface TFPI contributes to the overall anticoagulant profile of DX-9065a will require further investigation. PMID- 15815882 TI - Perioperative anticoagulation for patients with mechanical heart valves: a survey of current practice. AB - BACKGROUND: Patients with mechanical heart valves (MHV) require temporary interruption of warfarin when undergoing invasive procedures. Current guidelines addressing this subject are discordant because there is no high quality evidence to support any single management strategy. We tested the hypothesis that there is significant practice variation amongst clinicians caring for patients with MHV who require temporary cessation of their warfarin therapy. METHODS: A survey describing 4 hypothetical patients with mechanical heart valves was distributed to all clinicians attending an anticoagulation specialty meeting. For each scenario, the attendee was given several choices for preoperative and postoperative anticoagulation management. Information about each respondent's profession, specialty and the frequency with which they make perioperative anticoagulation recommendations was also collected. RESULTS: Three hundred twenty four of 650 surveys were returned. In each of the case scenarios, a majority of respondents selected subcutaneous low molecular weight heparin (LMWH) or subcutaneous unfractionated heparin (UH) as the preferred pre- and postoperative anticoagulant. Significant variation in practice was noted: for none of the questions was a single strategy selected by greater than 80% of respondents. CONCLUSION: Expert clinicians differ in their perioperative management strategies for patients with MHV who require interruption of warfarin. Although subcutaneous LMWH/UH was the treatment of choice in all scenarios, the lack of consensus found in our survey highlights the need for randomized controlled clinical trials of peri-procedural anticoagulant therapy. This survey of anticoagulation experts reveals that there is significant practice variation in scenarios where temporary interruption of warfarin is necessary in patients with mechanical heart valves. Despite discordant guidelines and a lack of high-quality data to support any strategy, a majority of the respondents surveyed would use low molecular weight heparin (or subcutaneous unfractionated heparin) to anticoagulate patients with mechanical heart valves during the peri-operative period. PMID- 15815883 TI - Budd-Chiari syndrome associated with visceral leishmaniasis and factor V Leiden mutation. AB - We here report a case of subacute Budd-Chiari syndrome (BCS) related to Factor V Leiden (FVL) mutation in the presence of visceral leishmaniasis. A 17-year-old man was admitted to hospital because of abdominal pain, pretibial edema and fever. The clinical picture of BCS had been developed within several months. BCS was diagnosed by radiographic examination. On DNA analysis, a heterozygote Arg506Gln mutation in the factor V gene was found. Histological examination of the bone marrow showed intracellular leishmania amastigotes. Despite appropriate treatment patient's clinical condition deteriorated rapidly and died with multiorgan failure. FVL mutation is the most common procoagulant disorder and account for many cases of BCS. This case report demonstrates that in addition to duration and severity of the disease accompanying conditions including infections are prognostically significant for the outcome of this potentially lethal disease. PMID- 15815884 TI - Pregnancies and oral contraceptive therapy in severe (homozygons) FXII deficiency: a study in 12 patients and review of the literature. AB - Twelve women with severe Factor XII (FXII) deficiency were under observation for an average period of about 16 years. During this time, these women had 19 pregnancies without any bleeding or thrombotic complications. The evaluation of the literature has shown that three patients manifested deep vein thrombosis during pregnancy. Five women also showed mild bleeding at delivery . The significance of these findings is not clear since thrombotic and bleeding complications may occur occasionally even in normal women. Five of our patients took oral contraceptive therapy during their fertile life for a variable period of time (2-10 years). No thrombosis was noted in any of these patients. From the scanty data gathered, in this respect, from the literature, it was shown that only three women with severe FXII deficiency took oral contraceptives and no thrombosis was noted. Altogether these results seem to indicate that the FXII deficiency does not play any significant role in the pathogenesis of bleeding and of thrombotic complications in pregnancy. However, the occurrence of deep vein thrombosis in 3 out of the 64 patients for whom sufficient data could be gathered indicates the need for further studies. This is more so if one considers that 3 out of the 6 cases of venous thromboses described altogether in the literature for females with severe FXII deficiency occurred during pregnancy or puerperium. PMID- 15815885 TI - Partial splenic embolization in myelodysplastic syndrome associated with immune thrombocytopenia. AB - BACKGROUND: Currently, all treatments for patients with myelodysplastic syndrome (MDS) are still experimental. Supportive care, including treatment of infection and transfusion of blood and blood-related products, remains the most important aspect of therapy in these cases. This article discusses the results of partial splenic embolization in a patient who had MDS with refractory anemia. METHODS: The patient was placed under observation and was given supportive treatment. In addition to MDS with refractory anemia, there were management difficulties related to severe thrombocytopenia after alloimmunization from multiple blood transfusions. Therefore it is thought that partial splenic embolization might be an alternative treatment for the management thrombocytopenia. RESULTS: Although the patient did not live long because of the sepsis and ileus, hematologic response was obtained. CONCLUSION: The observation during the treatment in this case suggests that partial splenic embolization may be a valid new alternative for treating thrombocytopenia due to alloimmunization. PMID- 15815887 TI - [Peranal passage of a surgical sponge: fortunate outcome of an intraoperative oversight]. AB - Four years after cholecystectomy and recurrent epigastric pain, a surgical sponge passed per anum in a 39-year-old man. Diagnostic procedures including clinical and laboratory examinations, ultrasound, coloscopy, and abdominal CT did not reveal any noteworthy residual changes by the foreign body. After 4 years in situ, the surgical sponge showed no alterations in structure. The representatives of the hospital concerned had to satisfy the patient's claims for damages. Pathology, technical diagnostic methods, and prognosis in cases of retained gauzes are presented. It is essential to count instruments and towels and advisable to use only single sponges in forceps in order to prevent intraoperative loss. PMID- 15815888 TI - [Therapeutic options for epithelial skin tumors. Actinic keratoses, Bowen disease, squamous cell carcinoma, and basal cell carcinoma]. AB - There has been worldwide a significant rise in the incidence of epithelial skin tumors and their precursors in the past years with an increased number of younger patients affected. The risk factors are identified. The choice of the appropriate treatment for each individual is crucial. Major consideration include high cure rate, low long-term recurrence rate and few side effects. In the following article different therapeutic approaches for actinic keratoses, Bowen's disease, basal cell carcinoma and squamous cell carcinoma are presented and analysed. PMID- 15815886 TI - [Preoperative evaluation and perioperative management of patients with increased cardiovascular risk]. AB - Due to the increasing age in western countries, combined with high rates of major surgical interventions in high-risk patients, perioperative reduction of cardiovascular complications becomes increasingly more important for perioperative physicians. After identifying patients with increased perioperative risk, specific interventions need to be considered to reduce their risk for cardiovascular complications, either by perioperative medical therapy or specific treatment options (e.g. coronary intervention). Several trials have demonstrated an effect of perioperative beta-blocker-therapy in reducing cardiovascular complications among high-risk patients. Additionally, several monitoring techniques are effective in detecting cardiovascular complications. Nevertheless, it remains unclear whether they are associated with a measurable improvement of outcome. Based on the ACC/AHA-guidelines, the present review describes a stepwise approach to surgical patients to identify perioperative risks, based on specific patient related risk factors, the kind of surgery and on the specific setting (emergency versus elective surgery). In addition, strategies to reduce perioperative cardiovascular complications are discussed. PMID- 15815890 TI - [The value of clinical infectology]. AB - Infectious diseases specialists are involved in patient care mostly regarding patients with severe infections in complex situations. Morbidity and mortality of severe infections such as sepsis has been significantly reduced during past years and decades. Prognosis of severe infections is dependent upon timely and correct diagnosis and appropriate therapy. The diagnostic and therapeutic approach is often invasive and interdisciplinary. Importantly, thoughtful use of antibiotics may increase its effectiveness, reduce antibiotic use and occurrence of bacterial resistance and increase cost effectiveness. Hence, antibiotic management and stewardship has become more and more the focus of work. This article highlights the clinical work of infectious diseases specialists in an interdisciplinary setting and describes there value to patients and hospitals. PMID- 15815889 TI - [Health economic aspects of psoriasis therapy. Is inpatient topical treatment of plaque-type psoriasis in this age of biologics still appropriate from both medical and economic viewpoints?]. AB - BACKGROUND: We tried to clarify whether the treatment of plaque psoriasis as an inpatient with dithranol and narrow band UV-B 311 nm is still appropriate and economical when compared to biologics such as efalizumab. PATIENTS AND METHODS: In a 15 month period, all patients admitted to the hospital after unsuccessful outpatient treatment of psoriasis were evaluated prospectively according to medical and economical criteria. RESULTS: PASI-score was reduced by 89% in 74 patients. The direct cost to improve one in-patient by 75% in PASI score (PASI 75) was 4342.27 EUR; the total costs for employed patients were 7441.81 EUR. Direct costs of treatment with efalizumab would come to 15048.89 EUR per patient with PASI-75 in a 12 weeks period. CONCLUSION: Topical inpatient therapy for plaque psoriasis is well tolerated and a less expensive option than biologics in those who fail ambulatory treatment. PMID- 15815891 TI - [Sense and nonsense in antibiotic therapy for respiratory tract infections]. AB - Whether an antibiotic successfully eradicates pathogens depends on the pathogens involved, on pharmacokinetics and bioavailability in the target tissue, and on the antimicrobial resistance of the pathogen. Other determinants are drug interactions, individual risk factors, age and compliance with respect to correct dosage and duration of therapy. In many cases, antimicrobial therapy is begun on an empirical basis, because the responsible pathogen can be identified in only half of all respiratory infections. The eradication of the pathogen has to be the first aim if treatment is to be curative and the development of resistance prevented. Long-term prevention of antimicrobial resistance will require a more critical prospective evaluation of the prescription of antibiotics. This paper considers rational and irrational measures in the antimicrobial therapy of respiratory infections. PMID- 15815892 TI - [CT diagnosis in acute cerebral ischemia]. AB - The advent of new MRI techniques such as perfusion- (PWI) and diffusion- (DWI) weighted imaging has improved diagnostic imaging in stroke. However, CT scanners are more widely available and less expensive than MRI scanners and are often located in the emergency departments even of smaller community hospitals. Topic of this article is CT-based diagnosis of patients with hyperacute ischemic stroke. In hyperacute stroke, a multiparametric CT protocol allows a comprehensive diagnosis by combining non-contrast enhanced CT (NECT), perfusion CT (PCT), and CT angiography (CTA). PCT can render important information about the hypoperfused brain tissue, CTA provides further important information about the vessel status. When stroke MRI is not available, multiparametric stroke CT can give nearly equivalent information, and can help to identify patients for thrombolytic therapy. PMID- 15815893 TI - [The radiology of opportunistic pneumonias. I: Epidemiological, laboratory and clinical background]. AB - In the guidelines of the German Society of Pneumology on diagnosis and therapy of opportunistic pneumonias, chest x-ray is listed as the basic diagnostic method in congenital and acquired immunodepression. In case of discrepancy between radiographic and the clinical findings or in cases of bilateral infiltrates, infection refractory to therapy or a difficult course in patients requiring artificial ventilation, a CT, or if necessary, ultrasound or MRI should be carried out. Cross-sectional imaging allows more precise assessment of the radiological pattern, estimation of the degree of severity (number of infiltrated segments) and detection of complications (pleural effusion, empyema, thorax wall infiltration). Clinical and laboratory parameters, bacteriological and serological examinations as well as information on the underlying immunocompromising factors must be taken into account in the differential diagnosis. The radiographic finding is an important diagnostic parameter which is used in the determination of the degree of severity of the pneumonia. The pattern of findings is one of rationales for the use of antibiotic therapy. In the first part of this contribution the epidemiological, laboratory and clinical background of the diagnosis of opportunistic pneumonias is discussed. PMID- 15815894 TI - [From loadstone to magnetic resonance tomography. Historical remarks on magnetism]. AB - This contribution offers a scientific historical view of the discovery, research, and medical use of magnetic phenomena. The milestones of the scientific findings beginning with the very first account of a classic magnetic force and progressing through to the new age discoveries of the magnetic resonance phenomenon with its subsequent use in medicine in the form of magnetic resonance tomography are presented. PMID- 15815895 TI - Randomised controlled trial comparing cisatracurium and vecuronium infusions in a paediatric intensive care unit. AB - OBJECTIVE: To evaluate and compare the efficacy, infusion rate and recovery profile of vecuronium and cisatracurium continuous infusion in critically ill children requiring mechanical ventilation. DESIGN AND SETTING: Prospective, randomised, double-blind, single-centre study in critically ill children in a paediatric intensive care unit in a tertiary children's hospital. METHODS: Thirty seven children from 3 months to 16 years old (median 4.1 year) were randomised to receive either drug; those already receiving more than 6 h of neuromuscular blocking drugs were excluded. The Train-of-Four (TOF) Watch maintained neuromuscular blockade to at least one twitch in the TOF response. Recovery time was measured from cessation of infusion until spontaneous TOF ratio recovery of 70%. RESULTS: The cisatracurium infusion rate in nineteen children averaged 3.9+/ 1.3 microg kg(-1) min(-1) with a median duration of 63 h (IQR 23-88). The vecuronium infusion rate in 18 children averaged mean 2.6+/-1.3 microg kg(-1) min(-1) with a median duration of 40 h (IQR 27-72). Median time to recovery was significantly shorter with cisatracurium (52 min, 35-73) than with vecuronium (123 min, 80-480). Prolonged recovery of neuromuscular function (>24 h) occurred in one child (6%) on vecuronium. CONCLUSIONS: Recovery of neuromuscular function after discontinuation of neuromuscular blocking drug infusion in children is significantly faster with cisatracurium than vecuronium. Neuromuscular monitoring was not sufficient to eliminate prolonged recovery in children on vecuronium infusions. PMID- 15815896 TI - Enteral donor pre-treatment with ursodeoxycholic acid protects the liver against ischaemia-reperfusion injury in rats. AB - Liver donor pre-treatment with ursodeoxycholic acid (UDCA) may protect against injury during transplantation. In the present study we evaluated whether enteral administration of UDCA has an effect on bile flow and protects the liver from injury related to transplantation. Wistar rats were used in liver perfusion (LP) and transplantation (LTx) models. Rats were enterally administered UDCA (800 mg/kg) 3 h before cold perfusion. In LP, bile flow and bile acid composition were analysed. In LTx, serum ALT and liver histology were analysed. LP showed biliary UDCA enrichment up to 36+/-13% in pre-treated rats, causing higher bile flow (P = 0.026) compared with control rats. LTx showed lower ALT and TUNEL positive hepatocytes in the UDCA group (P < 0.02 and P < 0.05). In conclusion, augmented bile salt-dependent bile flow is preserved in the liver after cold storage. Enteral donor pre-treatment with UDCA protects the liver against ischaemia reperfusion injury. PMID- 15815899 TI - Investigation of gettering effects in CZ-type silicon with SIMS. AB - Ion implantation is a well-known standard procedure in electronic device technology for precise and controlled introduction of dopants into silicon. However, damage caused by implantation acts as effective gettering zones, collecting unwanted metal impurities. This effect can be applied for "proximity gettering" reducing the concentration of impurities in the active device region. In this study the consequences of high-energy ion implantation into silicon and of subsequent annealing were analysed by means of secondary ion mass spectrometry (SIMS). Depth profiles were recorded of such impurities as copper, oxygen and carbon to obtain information about their gettering behaviour. The differences in impurities gettering behaviour were studied as a function of the implanted ions, P and Si, of the implantation dose and annealing time at T = 900 degrees C. Besides impurities gettering at the mean projected range (Rp) of implanted ions, Rp-effect, defects at around half of the projected ion range, Rp/2-effect, and even in some cases beyond Rp, trans-Rp-effect, have also been found to be effective in gettering of material impurities. PMID- 15815900 TI - Determination of methylmercury fluxes across the air-water and air-soil interfaces by gas chromatography with electron capture detection. AB - A method for the determination of methylmercury (MeHg) fluxes across the air water and air-soil interfaces was developed using an in situ chamber. The MeHg in the air coming out of the chamber was captured by a column containing sulfhydryl cotton fiber adsorbent. MeHg was then desorbed from the column by using 2 mol L( 1) HCl. The MeHg in the effluent was extracted with benzene, and determined by gas chromatography with electron capture detection. Finally, the MeHg flux was calculated using the chamber. The method was applied to simulated experiments, and the results showed that the MeHg fluxes in the air-water system were higher than those in the air-soil-water system. The method was also successfully applied to the field measurements of an environment polluted by a chemical factory, and the results showed that the MeHg fluxes across the air-soil and air-water interfaces were 0.21-3.09 and 0.14-0.79 ng m(-2) h(-1), respectively. The method will be a useful tool in the environmental study of MeHg. PMID- 15815901 TI - Optic nerve infiltration by acute lymphoblastic leukemia: MRI contribution. AB - We describe the clinical presentation and imaging features of a patient with acute lymphoblastic leukemia (ALL) that was complicated by optic nerve infiltration. The clinical and diagnostic characteristics of this complication must be recognized so that optimal therapy can be started to prevent blindness. MR imaging is useful in early detection and should be performed in any leukemic patient with ocular complaints, even during remission. PMID- 15815902 TI - Imaging characteristics of primary cranial Ewing sarcoma. AB - BACKGROUND: Ewing sarcoma accounts for 10-15% of all childhood malignant bone tumours and is second in prevalence to osteosarcoma. The skull bones are an unusual site of origin of primary Ewing sarcoma in children. Previous reports concentrate on the neurosurgical aspects and relatively good outcome compared to other bone tumours of the calvarium. Reported cases mainly describe the imaging characteristics on CT. OBJECTIVE: To describe the MRI and CT features of primary cranial Ewing sarcoma. MATERIALS AND METHODS: The neuroimaging of three cases of primary cranial Ewing sarcoma are reviewed. RESULTS: Our three cases show an extra-axial mass that is high attenuation on CT and low signal on T2-weighted MRI. Haemorrhagic components, dural extension and contrast enhancement are also characteristic features. CONCLUSION: CT attenuation and magnetic resonance signal characteristics reflect sheets of densely packed cells seen in Ewing sarcoma. PMID- 15815903 TI - Re-dislocation following open reduction for developmental dysplasia of the hip. AB - We have reviewed 17 patients (18 hips) who required repeat open reduction for recurrent or persistent dislocation after a previous attempt at open reduction for developmental dysplasia of the hip (DDH). The mean follow-up was 5.5 (3-15) years. At the review, five patients were limping and had limited hip motion but no pain. Six hips were classified as Severin class III or more, and avascular necrosis was evident in 11. We suggest that technical failure is usually the cause for re-dislocation in unilateral cases, as we discovered at repeat open reduction in five of six patients with unilateral dislocation. The biological behaviour of bilateral dislocations may in some way be different since in nine of 11 patients with bilateral dislocations, no obvious cause could be attributed. The outcome is usually poor. PMID- 15815904 TI - The effect of traction on compartment pressures during intramedullary nailing of tibial-shaft fractures. A prospective randomised trial. AB - Our aim was to study the effect of traction on the compartment pressures during intramedullary nailing of closed tibial-shaft fractures. Thirty consecutive patients with Tscherne C1 fractures were randomised into two groups. Sixteen patients underwent intramedullary nailing of the tibia with traction and 14 patients without traction. Compartment pressures were measured before the application of traction or commencement of the procedure and at the end of the procedure. The data collected was analysed using Student's t test. There was no statistically significant difference (p>0.05) in the pre-operative mean compartment pressures for both groups. The mean post-operative measurements were higher in all four compartments in the traction group (p<0.05). None of the pressures reached the critical level. These results show that traction as an aid unnecessarily increases compartment pressures. PMID- 15815905 TI - Compression nailing for posttraumatic rotational femoral deformities: open versus minimally invasive technique. AB - Between January 1996 and December 1999, we performed 30 derotational osteotomies with compression nailing in 29 patients. In 18 cases (group 1), we used an intramedullary saw (minimally invasive technique), and in 12 cases (group 2), we used a conventional open technique. Follow-up included clinical, conventional radiological, and computer tomographical assessment. The mean angle of derotation was 28.6+/-12.3 degrees in group 1 and 27.6+/-10.7 degrees in group 2. The postoperative mean rotational deviation between left and right side was 7.9+/-6.7 degrees in group 1 and 6.6+/-4.4 degrees in group 2. There were five postoperative complications: two delayed unions, two insufficient corrections, and one infection. There was no significant difference between the groups. When using the minimally invasive technique, we recommend the derotation angle to be marked with Schanz screws instead of Kirschner wires, as soft-tissue resistance may lead to bending of these. PMID- 15815906 TI - Diffuse venous thromboemboli associated with IVIg therapy in the treatment of streptococcal toxic shock syndrome: case report and review. AB - Streptococcal toxic shock syndrome (STSS) is a severe invasive disease with a 40 80% mortality rate. Inflammatory cytokines induced by streptococcal pyrogenic exotoxins (SPEs) produce the clinical manifestations of a flu-like syndrome, followed by high fevers and multiorgan failure. Previously published reports have described the use of intravenous immunoglobulin (IVIg) as adjunctive treatment for STSS. However, concerns have been raised about the thromboembolic complications of this therapy. We report a severe case of STSS treated with two adjunctive courses of IVIg complicated by severe bilateral pulmonary thromboemboli. To our knowledge, this is the only reported case of thromboemboli associated with IVIg for STSS. The results of this case support the cautious use of IVIg for STSS and demonstrate the need for controlled trials to determine the appropriate timing, dosage, and course of treatment. PMID- 15815907 TI - Expression of interferon-gamma and tumor necrosis factor-alpha in bone marrow T cells and their levels in bone marrow plasma in patients with aplastic anemia. AB - Immune-mediated stem cell damage has been postulated to be responsible for disease initiation and progression in aplastic anemia (AA). It is hypothesized that T lymphocytes play a major role in destroying the bone marrow (BM) stem cells of AA patients by infiltrating the BM and secreting excessive levels of anti-hematopoietic cytokines, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha). We undertook this study to assess the pathogenic significance of anti-hematopoietic cytokines such as IFN-gamma and TNF-alpha in BM T cells and plasma of AA patients. Significantly elevated levels of IFN-gamma and TNF-alpha were found in the BM plasma of AA patients compared to controls (p=0.05 and 0.006, respectively). Intracellular IFN-gamma and not TNF-alpha in BM CD3+ T cells of AA patients was significantly higher compared to controls (p=0.04 and p=0.2, respectively). A follow-up analysis of expression of these cytokines in BM T cells and their levels in BM plasma in five AA patients before and 180 days (6 months) after antithymocyte globulin (ATG) and cyclosporin A (CsA) therapy showed a decline 180 days after therapy compared to pre-therapy. We thus conclude that increased production of both IFN-gamma and TNF-alpha in the BM may contribute to disease pathogenesis in AA and ATG therapy may induce hematological remission by suppressing the elevated levels of IFN-gamma and TNF-alpha in AA BM. PMID- 15815908 TI - Dysphagia revealing esophageal involvement by non-Hodgkin's lymphoma. PMID- 15815909 TI - Spontaneous rupture of the spleen in a patient with B-cell acute lymphoblastic leukaemia. PMID- 15815910 TI - 3D motion adapted gating (3D MAG): a new navigator technique for accelerated acquisition of free breathing navigator gated 3D coronary MR-angiography. AB - This study aimed to evaluate the influence of a new navigator technique (3D MAG) on navigator efficiency, total acquisition time, image quality and diagnostic accuracy. Fifty-six patients with suspected coronary artery disease underwent free breathing navigator gated coronary MRA (Intera, Philips Medical Systems, 1.5 T, spatial resolution 0.9x0.9x3 mm3) with and without 3D MAG. Evaluation of both sequences included: 1) navigator scan efficiency, 2) total acquisition time, 3) assessment of image quality and 4) detection of stenoses >50%. Average navigator efficiencies of the LCA and RCA were 43+/-12% and 42+/-12% with and 36+/-16% and 35+/-16% without 3D MAG (P<0.01). Scan time was reduced from 12 min 7 s without to 8 min 55 s with 3D MAG for the LCA and from 12 min 19 s to 9 min 7 s with 3D MAG for the RCA (P<0.01). The average scores of image quality of the coronary MRAs with and without 3D MAG were 3.5+/-0.79 and 3.46+/-0.84 (P>0.05). There was no significant difference in the sensitivity and specificity in the detection of coronary artery stenoses between coronary MRAs with and without 3D MAG (P>0.05). 3D MAG provides accelerated acquisition of navigator gated coronary MRA by about 19% while maintaining image quality and diagnostic accuracy. PMID- 15815911 TI - Multidetector computed tomography enteroclysis (MDCT-E) with neutral enteral and IV contrast enhancement in tumor detection. AB - The aim of our study was to evaluate the reliability of MDCT-enteroclysis (MDCT E), an emerging technique of small bowel examination that combines the advantages of MDCT examination of the abdomen and the enteral volume challenge of enteroclysis, in evaluation of small bowel neoplasms. In our institutions, MDCT-E was used to study 456 patients (age range 21-84 years, mean 53 years) admitted for suspicion of small bowel disease. All examinations were done on multichannel CT units, 129 on a 16-slice scanner and 327 on a four-slice scanner. Post processing and multiplanar reformatting and interpretation were performed on dedicated workstations. After adequate gastrointestinal preparation and naso enteric intubation, small bowel was distended by methylcellulose infused by a peristaltic pump. One volumetric MDCT acquisition was obtained after 120-130 ml of intravenous contrast medium. Multiplanar reformatting was based on the image reconstruction parameters from 3 to 4 mm. Forty-five small bowel neoplasms were found; in the remaining cases, 223 Crohn's diseases and 149 other abnormalities were detected. All findings were confirmed by surgery, endoscopy or clinical follow-up. In our experience, MDCT-E with neutral enteral and IV contrast seems to be a reliable method in the diagnosis of small bowel neoplasms. PMID- 15815912 TI - [Dose-effect relation in revision surgery for consecutive strabismus divergens in adults]. AB - BACKGROUND: The aim of this study was to establish a dose-effect relationship between the reduction of the angle of deviation and the millimeters of the reoperation performed in patients with consecutive exotropia. PATIENTS AND METHODS: A total of 46 patients who had undergone reoperation for consecutive exotropia (group 1: advancement of the former recessed medial rectus muscle and group 2: additional rececession of the lateral rectus muscle) were reexamined. RESULTS: The dose-effect relationship was 2 degrees /mm in group 1 and1.6 degrees /mm far distance and 1.8 degrees /mm near in group 2. Within the first 4 months after the reoperation we found an exodrift of up to 6 degrees , which remained stable; 98% of our patients reported that their quality of life had improved. CONCLUSION: In cases of consecutive exotropia we consider the postoperative exdodrift in the planing of our reoperation. During the ensuing years patients can expect stability of the results achieved by the reoperation. PMID- 15815913 TI - [Vision loss after contusio bulbi]. PMID- 15815914 TI - [New tumor onset of the plica semilunaris in advanced age]. PMID- 15815915 TI - [Complicated course in a patient with chronic conjunctivitis]. PMID- 15815916 TI - TCR gamma delta (+) T-cell response to streptococcal antigens in patients with psoriasis. PMID- 15815918 TI - An Arabidopsis mutant that is resistant to the protoporphyrinogen oxidase inhibitor acifluorfen shows regulatory changes in tetrapyrrole biosynthesis. AB - Several Arabidopsis mutants of the ecotype Dijon were isolated that show resistance to the herbicide acifluorfen, which inactivates protoporphyrinogen oxidase (PPOX), an enzyme of tetrapyrrole biosynthesis. This enzyme provides protoporphyrin for both Mg chelatase and ferrochelatase at the branchpoint, which leads to chlorophyll and heme, respectively. One of the mutations, aci5-3, displays semidominant inheritance. Heterozygous progeny showed yellow-green leaves, while the homozygous seedlings were white and inviable, but could be rescued by supplementing the medium with sugar. Interestingly, the expression of neither of the two forms of PPOX was altered in the mutant, but the rate of synthesis of 5-aminolevulinate, the precursor of all tetrapyrroles, was drastically reduced. Genetic mapping revealed the mutant locus is closely linked to the ch42 marker, which is itself located in the CHLI-1 gene which codes for one of the three subunits of Mg chelatase. The cs mutant also shows a defect in this gene, and test for allelism with aci5-3 confirmed that the two mutations are allelic. Sequencing of the wild type and aci5-3 alleles of CHLI-1 revealed a single base change (G718A), which results in a D240N substitution in the CHLI-1 protein. In the homozygous aci5-3 mutant no CHLI-1 RNA or protein could be detected. Strikingly, CHLH and CHLI-2 transcripts were also absent. This indicates the existence of a feedback-regulatory mechanism that inactivates the genes encoding certain Mg chelatase subunits. The basis for the semidominant inheritance pattern and the relationship between herbicide resistance and modified gene expression is discussed. PMID- 15815917 TI - Effects of waterproof covering on hand immersion tests using water at 10 degrees C, 12 degrees C and 15 degrees C for diagnosis of hand-arm vibration syndrome. AB - OBJECTIVES: To compare effects of waterproof covering on finger skin temperature (FST) and subjective hand pain during immersion tests using cold water at 10 degrees C, 12 degrees C and 15 degrees C. In the (Draft International Standard) of the International Organization for Standardization (ISO/DIS 14835-1), a water temperature of 12 degrees C and use of water covering are proposed. METHODS: Six healthy male subjects took part in the immersion tests and immersed both hands into water at 10 degrees C, 12 degrees C and 15 degrees C for 5 min, repeatedly, with waterproof covering (polyethylene gloves) or without (bare hands). The FST data from middle fingers and subjective pain scores for hand pain were analyzed. Furthermore, the test with water at 12 degrees C was repeated to assess the repeatability of the test. RESULTS: The glove and water temperature factors for FST were significant at every minute from 1 min during immersion up to 2 min after recovery, showing higher values for waterproof covering than for bare hands and showing lowest values for water temperature of 10 degrees C and highest for 15 degrees C. The glove and water temperature factors for subjective pain score were significant at the 1-min and 2-min points during immersion, showing lower scores for waterproof covering than for bare hands and showing highest scores for water temperature of 10 degrees C and lowest for 15 degrees C. The results of the first and second tests using water of 12 degrees C showed no systematic difference in FST and hand pain between the tests, with a few exceptions. CONCLUSIONS: Subjective pain during the cold immersion test with polyethylene gloves and water at 12 degrees C can be reduced, while the differences in FST between water temperatures of 10 degrees C and 12 degrees C were small or not apparent at some points during immersion and recovery. The test also seems to be suitable for repeatability. Further investigation on hand-arm vibration syndrome (HAVS) patients to validate the use of the immersion test with gloves to obtain sufficient data for diagnostic value is required. PMID- 15815921 TI - Extraction of extraradical arbuscular mycorrhizal mycelium from compartments filled with soil and glass beads. AB - This study presents a novel method for the extraction and quantification of extraradical mycelium (ERM) of arbuscular mycorrhizal fungi (AMF) from a substrate that simulates soil better than previously used artificial growth media. Fungal compartments were constructed from small net pots with a latticed wall and filled with a mixture of glass beads and 40 microm wet sieved soil. The net pots were surrounded by a 30-microm mesh membrane through which hyphae but not roots could grow. They were inserted into soil where a Glomus intraradices (BEG 110) colonized potato plant was growing. The ERM that had grown out from roots through the membrane was successfully collected and quantified after harvest by washing out the soil/glass bead mixture through a sieve with a mesh width of 40 microm. Concentrations of P, Zn, Cu and Mn in the AMF ERM were analysed. PMID- 15815922 TI - Effect of power density of curing unit, exposure duration, and light guide distance on composite depth of cure. AB - This in vitro study compared the depth of cure obtained with six quartz tungsten halogen and light-emitting diode curing units at different exposure times and light tip-resin composite distances. Resin composite specimens (Tetric Ceram, A3; diameter 4 mm, height 6 mm) were exposed from 0-, 3-, and 6-mm distance. The curing units (200-700 mW/cm2) were used for standard (20 and 40 s), pulse-delay mode (initial exposure of 3 s at 200 mW/cm2, followed by a resting period of 3 min and a final exposure of 10 or 30 s at 600 mW/cm2), or soft-start curing (40 s; exponential ramping). Curing depth was determined by measurement of Wallace hardness for each half millimeter starting at 0.5 mm from the top surface. For each specimen, a mean H(W) value was calculated from the H(W) values determined at the depths of 2.0 mm and less (0.5, 1.0, 1.5, and 2.0 mm, respectively). The depth of cure for each specimen was found by determining the greatest depth before an H(W) value exceeding the minimal H(W) value by 25% occurred. For all curing units, an increase in exposure time led to significantly higher depth of cure. Increasing the light tip-resin composite distance significantly reduced the depth of cure. With a light tip-resin composite distance of 6 mm, median values of depth of cure varied between 2.0 and 3.5 mm following a 20-s (or 3+10 s) exposure and between 3.0 and 4.5 mm following a 40-s (or 3+30 s) exposure. The composite situated above the depth of cure value cured equally well with all curing units. At both exposure times, Luxomax resulted in the significantly lowest depth of cure, and Astralis 7 yielded significantly higher depth. At both exposure times, a significant linear correlation was found between the determined power densities of the curing units and the pooled depth of cure values obtained. It seems that for the resin composite tested, the recommended exposure time of 40 s per 2-mm increment may be reduced to 20 s, or that increments may be increased from 2 to 3.5 mm. It may be that the absolute values of depth of cure found are material specific, but we believe that the relationships found between curing units, between exposure times, and between light guide distances are universal. PMID- 15815923 TI - In vivo interfacial adaptation of class II resin composite restorations with and without a flowable resin composite liner. AB - The aim of this study was to evaluate in vivo the interfacial adaptation of class II resin composite restorations with and without a flowable liner. In 24 premolars scheduled to be extracted after 1 month, 48 box-shaped, enamel-bordered class II cavities were prepared and restored with a flowable liner (FRC, Tetric Flow/Tetric Ceram/Syntac Single-Component) or without (TRC), cured with three different curing modes: soft start and 500- or 700-mW/cm2 continuous irradiation. Interfacial adaptation was evaluated by quantitative scanning electron microscopic analysis using replica method. Gap-free adaptation in the cervical enamel (CE) was observed for FRC and TRC in 96.2 and 90.2%, for the dentin (D) in 63.6 and 64.9%, and for occlusal enamel (OE) in 99.7 and 99.5%, respectively. The difference between the two restorations was not statistically significant (ns). Significant better adaptation was observed for OE than CE and D (p<0.01), and for CE than D (p<0.01). Gap-free adaptation with the soft-start and 500- and 700 mW/cm2 continuous-curing modes was observed for CE: 88.7%, 92.7%, 97.9% (ns); OE: 99.8%, 98.7%, 100% (ns); and D: 64.0%, 63.9%, and 64.6% (ns), respectively. It can be concluded that neither the use of flowable resin composite liner nor the curing mode used influenced the interfacial adaptation. PMID- 15815924 TI - Permutation tests for the correlation among genetic distances and measures of heterosis. AB - It is often found that heterosis tends to increase with genetic distance of the parents, though the correlation is not usually very close. It is therefore important to test the null hypothesis that the correlation is zero. The present work shows that standard procedures tend to yield too liberal tests, owing to the lack of independence among genetic distances and among heterosis estimates. A valid alternative is to use a permutation test, which was first suggested by Mantel [(1967) Cancer Res 27: 209--220). This test is well-known among plant breeders and geneticists, who often use it to test the correlation among two distance matrices. Its use is not restricted to the comparison of distance matrices. This is demonstrated in the present work, using two published datasets on marker-based genetic distances of maize inbreds or populations and heterosis of their crosses. It is shown that the test is also applicable in the presence of missing data. PMID- 15815925 TI - Identification and molecular tagging of a gene from PI 289824 conferring resistance to leaf rust (Puccinia triticina) in wheat. AB - Host-plant resistance is the most economically viable and environmentally responsible method of control for Puccinia triticina, the causal agent of leaf rust in wheat (Triticum aestivum L.). The identification and utilization of new resistance sources is critical to the continued development of improved cultivars as shifts in pathogen races cause the effectiveness of widely deployed genes to be short lived. The objectives of this research were to identify and tag new leaf rust resistance genes. Forty landraces from Afghanistan and Iran were obtained from the National Plant Germplasm System and evaluated under field conditions at two locations in Texas. PI 289824, a landrace from Iran, was highly resistant under field infection. Further evaluation revealed that PI 289824 is highly resistant to a broad spectrum of leaf rust races, including the currently prevalent races of leaf rust in the Great Plains area of the USA. Eight F1 plants, 176 F2 individuals and 139 F2:3 families of a cross between PI 289824 and T112 (susceptible) were evaluated for resistance to leaf rust at the seedling stage. Genetic analysis indicated resistance in PI 289824 is controlled by a single dominant gene. The AFLP analyses resulted in the identification of a marker (P39 M48-367) linked to resistance. The diagnostic AFLP band was sequenced and that sequence information was used to develop an STS marker (TXW200) linked to the gene at a distance of 2.3 cM. The addition of microsatellite markers allowed the gene to be mapped to the short arm of Chromosome 5B. The only resistance gene to be assigned to Chr 5BS is Lr52. The Lr52 gene was reported to be 16.5 cM distal to Xgwm443 while the gene in PI 289824 mapped 16.7 cM proximal to Xgwm443. Allelism tests are needed to determine the relationship between the gene in PI 289824 and Lr52. If the reported map positions are correct, the gene in PI 289824 is unique. PMID- 15815926 TI - Root response to Fusarium solani f. sp . glycines: temporal accumulation of transcripts in partially resistant and susceptible soybean. AB - Sudden death syndrome (SDS) of soybean is a complex of root rot disease caused by the semi-biotrophic fungus Fusarium solani f. sp. glycines (Fsg) and a leaf scorch disease caused by toxins produced by the pathogen in the roots. Development of partial rate-reducing resistance in roots to SDS was studied. The recombinant inbred line 23 (RIL23) that carried resistance conferred by six quantitative trait loci (QTL) derived from cultivars 'Essex' x 'Forrest' was compared to the susceptible cultivar Essex. Roots of RIL23 and its susceptible parent Essex were inoculated with Fsg. Transcript abundance (TA) of 191 ESTs was studied at five time points after inoculation. For most of the genes, there was an initial decrease in TA in the inoculated roots of both genotypes. By days 7 and 10 the inoculated roots of Essex failed to increase expression of the transcripts of defense-related genes. In RIL23 inoculated roots, the TA of 81 genes was increased by at least two-fold at day 3 (P=0.004), 88 genes at day 7 (P=0.0023) and 129 genes at day 10 (P=0.0026). A set of 35 genes maintained at least a two-fold higher abundance at all three time points. The increase in TA in RIL23 was in contrast to that observed in Essex where most of the ESTs showed either no change or a decreased TA. The ESTs with an increased TA had homology to the genes involved in resistance (analogs), signal transduction, plant defense, cell wall synthesis and transport of metabolites. Pathways that responded included the protein phosphorylation cascade, the phospholipase cascade and the phenolic natural products pathways, including isoflavone and cell wall synthesis. PMID- 15815927 TI - During attachment Phytophthora spores secrete proteins containing thrombospondin type 1 repeats. AB - Adhesion is a key aspect of disease establishment in animals and plants. Adhesion anchors the parasite to the host surface and is a prerequisite for further development and host cell invasion. Although a number of adhesin molecules produced by animal pathogens have been characterised, molecular details of adhesins of plant pathogens, especially fungi, are largely restricted to general descriptions of the nature of heterogeneous secreted materials. In this paper, we report the cloning of a gene, PcVsv1, encoding a protein secreted during attachment of spores of Phytophthora, a genus of highly destructive plant pathogens. PcVsv1 contains 47 copies of the thrombospondin type 1 repeat, a motif found in adhesins of animals and malarial parasites but not in plants, green algae or true fungi. Our results suggest that PcVsv1 is a spore adhesin and highlight intriguing similarities in structural and molecular features of host attachment in oomycete and malarial parasites. PMID- 15815928 TI - Mammalian Bax initiates plant cell death through organelle destruction. AB - Mammalian Bax is known to cause cell death when expressed in plants. We examined transgenic plants expressing both Bax and organelle-targeted green fluorescent protein to determine the cellular changes that occur during Bax-induced cell death. The mitochondria changed morphologically from being bacilli-shaped to being round, eventually becoming swollen. Mitochondria streaming also stopped. The chloroplasts lost membrane function and their contents leaked out, followed by the disruption of the vacuole. Light was not essential for Bax-induced ion leakage or organelle disruption. These results indicate that Bax induces temporal and spatial cell death events at the organelle level in the plant. A heterologous system, using Bax, would therefore be available to investigate cell death, which is commonly conserved in animals and plants. PMID- 15815929 TI - Transformation of a recalcitrant grain legume, Vigna mungo L. Hepper, using Agrobacterium tumefaciens-mediated gene transfer to shoot apical meristem cultures. AB - The efficiency of Vigna mungo L. Hepper transformation was significantly increased from an average of 1% to 6.5% by using shoot apices excised from embryonic axes precultured on 10 microM benzyl-6-aminopurine (BAP) for 3 days and wounded prior to inoculation in Agrobacterium tumefaciens strain EHA105 carrying the binary vector pCAMBIA2301, which contains a neomycin phosphotransferase gene (nptII) and a beta-glucuronidase (GUS) gene (gusA) interrupted by an intron. The transformed green shoots that were selected and rooted on medium containing kanamycin, and which tested positive for nptII gene by polymerase chain reaction, were established in soil to collect seeds. GUS activity was detected in whole T(0) shoots and T(1) seedlings. All T(0) plants were morphologically normal, fertile and the majority of them transmitted transgenes in a 3:1 ratio to their progenies. Southern analysis of T(1) plants showed integration of nptII into the plant genome. PMID- 15815930 TI - [Pain assessment in invasive diagnostic procedures. Comparison of an eleven-point numerical rating scale and a six-point verbal rating scale for pain measurement in bone marrow puncture]. AB - BACKGROUND AND OBJECTIVE: Pain measurement during diagnostic procedures is an accepted prerequisite for appropriate therapy. In this study, the agreement between rankings of pain intensity on a numerical and a verbal rating scale was analyzed. METHODS: Patients attending a haematological outpatient clinic who underwent bone marrow puncture were requested to assess pain intensity perceived during the procedure on a elen-point numerical and a six-point verbal rating scale. RESULTS: Analysis of patients' pain ratings in 263 bone marrow punctures showed a very good correlation between numerical and verbal rating scale (Spearman correlation coefficient 0,86). By means of a transfer instruction categories of the verbal rating scale were assigned to figures of the numerical rating scale (0 - no pain, 1 or 2 - mild p., 3, 4 or 5 moderate p., 6 or 7 severe p., 8, 9 or 10 - very severe and worst possible pain). The resulting transfer table showed a good agreement with a weighted kappa of 0.72 (95% confidence interval: 0.66-0.79). CONCLUSION: Both scales can be employed efficiently for pain assessment during diagnostic procedures. Verbal categories can be assigned to numerical values and vice versa numerical values to verbal categories. However, in view of the inter-individual variations it appears appropriate to re assure pain perception with each patient in order to avoid over- or undertreatment. PMID- 15815931 TI - Mutational analysis of EGFR and K-RAS genes in lung adenocarcinomas. AB - Both epidermal growth factor receptor (EGFR) and RAS gene mutations contribute to the development of non-small cell lung cancer (NSCLC). Because RAS is one of the downstream molecules in the EGFR signal transduction, the association between the somatic mutations of EGFR and RAS may be important in the pathogenesis of NSCLC . However, to date, such data are lacking. In this study, we analyzed the hotspot regions of K-RAS gene (codons 12, 13, 59 and 61) and EGFR gene (exons 18, 19 and 21) in 153 NSCLC tissue samples including 69 adenocarcinomas. Overall, we detected 30 EGFR mutations (19.6%) and 6 K-RAS mutations (3.9%) in the 153 NSCLCs. In the 69 adenocarcinomas, 26 EGFR mutations (37.7%) and six K-RAS mutations (8.7%) were detected. Of note, the 26 tumors with EGFR mutations did not harbor any K-RAS mutations, and the six tumors with K-RAS mutations did not harbor any EGFR mutations. Inverse relationship between K-RAS and EGFR mutations in the lung adenocarcinoma was statistically significant (P=0.046, chi2 test). As regards smoking history, EGFR mutation was significantly associated with never smoking history, whereas K-RAS mutation was significantly associated with smoking history. Our data suggest that mutations of EGFR and K-RAS genes might separately, but not cooperatively, contribute to lung adenocarcinoma pathogenesis, and that EGFR and K-RAS mutants could separately be anti-neoplastic targets in lung adenocarcinomas. PMID- 15815932 TI - Hepatic adenocarcinoma expressing inhibin in a young patient on oral contraceptives. AB - A case of primary hepatic carcinoma is reported, which occurred in a 24-year-old woman with a 10-year history of oral contraceptive use, and demonstrated unique morphologic and immunohistochemical features. The tumor was located in the left hepatic lobe, measured 14 cm at its widest, and showed histologic heterogeneity. The neoplastic cells were mostly arranged in trabecular and pseudoglandular growth patterns simulating hepatocellular carcinoma; however, in focal areas, small cystic, organoid and tubular patterns predominated. Immunohistochemical stains showed a phenotype consistent with biliary differentiation (positive staining for cytokeratin 7, cytokeratin 19, carcinoembryonic antigen and CA 19-9 antigen). The tumor cells were negative for markers that would be suggestive of hepatocytic or neuroendocrine differentiation. Interestingly, they were positive for inhibin, a protein that is known to be expressed in sex cord-stromal tumors of the ovary, trophoblastic neoplasms and adrenal cortical tumors, but not in hepatic tumors. However, no definite evidence of gonadal stromal, trophoblastic, or adrenocortical differentiation was identified on extensive immunohistochemical work-up. In conclusion, this unique case may represent a rare variant of cholangiocarcinoma expressing inhibin. PMID- 15815933 TI - Abdominal inflammatory masses mimicking neoplasia in children-experience of two centers. AB - Despite progress in modern imaging, some inflammatory masses are difficult to distinguish clinically from neoplastic processes. In such cases the pathology report has a great distinctive value, but even then the final diagnosis may be difficult to reach. Eight patients with abdominal tumors of inflammatory origin were treated in two institutions, the Department of Pediatric Surgery of the Medical University of Gdansk, Poland, and Helios Center of Pediatric Surgery in Berlin, Germany, during the last 10 years. Four tumors were located in the pelvis, two in the liver, and two in the colonic mesentery. Five of them were inflammatory pseudotumors (two subclassified as inflammatory fibrosarcoma), one had nonspecific inflammatory changes, one was diagnosed as idiopathic retroperitoneal fibrosis, and one was diagnosed as bacillary angiomatosis. All patients underwent surgical tumor biopsy, excisional in four and incisional in four. All but two children underwent macroscopically complete tumor excision (four primarily, two secondarily). In one case the tumor resolved with antibiotherapy. Surgery in retroperitoneal masses was often extensive and associated with significant complications because of invasive tumor growth. In conclusion, intraabdominal inflammatory lesions may closely mimic neoplasia in children. Clinical doubts result in repeated biopsies, and for this reason excisional biopsy should be preferred. In some cases, when excisional biopsy is not feasible due to invasive growth of the tumor, delayed complete mass excision should follow, despite occasional significant morbidity. The etiology and exact nature of inflammatory pseudotumors are still obscure, and it is unknown whether they represent inflammatory lesions or true neoplasia. PMID- 15815936 TI - Cardiovascular effects of anti-G suit inflation at 1 and 2 G. AB - We sought to determine to which pressure a full-coverage anti-G suit needs to be inflated in order to obtain the same stroke volume during a brief exposure to twice the normal gravity (2 G) as that at 1 G without anti-G suit inflation. Nine sitting subjects were studied at normal (1 G) and during 20 s of exposure to 2 G. They wore anti-G suits, which were inflated at both G-levels to the following target pressures: 0, 70, 140 and 210 mmHg. Stroke volume was computed from cardiac output, which was measured by rebreathing. Heart rate and mean arterial pressure at heart level were recorded. Inflation to 70 mmHg compensated for the decrease in stroke volume and cardiac output caused by hypergravity. Mean arterial pressure at heart level was comparable at 1 G and at 2 G and increased gradually and similarly with inflation (P<0.001) at both gravity levels. Thus, anti-G suits act by increasing both preload and afterload but the two effects counteract each other in terms of cardiac output, so that cardiac output at 2 G is maintained at its 1 G level. This effect is reached already at 70 mmHg of inflation. Greater inflation pressure further increases mean arterial pressure at heart level and compensates for the increased difference in hydrostatic pressure between heart and head in moderate hypergravity. PMID- 15815935 TI - Do blood cells mimic gene expression profile alterations known to occur in muscular adaptation to endurance training? AB - Exercise is known to upregulate mRNA synthesis for carnitine palmitoyl transferase1 (CPT1) and possibly also other mitochondrial carnitine acyltransferases in muscle tissue. The aim of this study was to test whether such an adaptation of oxidative metabolism in skeletal muscle is a systemic process and consequently, also affects other cells. Messenger RNA levels of five genes [carnitine palmitoyl transferases 1 and 2 (CPT1 and CPT2), carnitine acetyltransferase (CRAT), carnitine palmitoyltransferase 2 (CPT2), microsomal carnitine palmitoyltransferase (GRP58) and organic cation transporter (OCTN2)] were determined with quantitative real time polymerase chain reaction (PCR) in blood cells and in muscle biopsy samples from six cross country skiers before and 6 months after a high volume/low intensity exercise training, when training had elicited a significantly slower rate of lactate accumulation. Quantitative real time PCR showed that levels of mRNA in blood cells correlated significantly (CPT1B: P< 0.001) with those in muscle tissue from the same donors. After 6 months training, there was a 15-fold upregulation of CPT1B mRNA, a six to ninefold increase of CRAT mRNA, of CPT2 mRNA, GRP58 mRNA, and of OCTN2 mRNA. The observation of a concordant stimulation of CPT1, CPT2, CRAT, GRP58 and OCTN2 transcription in blood cells and muscle tissue after 6-month-endurance training leads the hypothesis of a common stimulation mechanism other than direct mechanical stress or local chemical environment, but rather humoral factors. PMID- 15815937 TI - The validity of predicting maximal oxygen uptake from a perceptually-regulated graded exercise test. AB - The purpose of this study was to assess the validity of predicting maximal oxygen uptake(VO(2max)) from sub-maximal VO(2) values elicited during a perceptually regulated exercise test. We hypothesised that the strong relationship between the ratings of perceived exertion (RPE) and VO(2) would enable VO(2max) to be predicted and that this would improve with practice. Ten male volunteers performed a graded exercise test (GXT) to establish VO(2max) followed by three sub-maximal RPE production protocols on a cycle ergometer, each separated by a period of 48 h. The perceptually-regulated trials were conducted at intensities of 9, 11, 13, 15 and 17 on the RPE scale, in that order. VO(2) and HR were measured continuously and recorded at the end of each 4 min stage. Individual's RPE values yielded correlations in the range 0.92-0.99 across the three production trials. There were no significant differences between measured VO(2max) (48.8 ml.kg(-1).min(-1)) and predicted VO(2) max values (47.3, 48.6 and 49.9 ml.kg(-1).min(-1), for trials 1, 2 and 3, respectively) when VO(2) max was predicted from RPE values of 9-17. The same was observed when VO(2max) was predicted using RPE 9-15. Limits of agreement (LoA) analysis on actual and predicted VO(2max) values (from RPE 9-17) were (bias+/-1.96xSDdiff) 1.5+/-7.3, 0.2+/-4.9 and -1.2+/-5.8 ml.kg(-1).min(-1), for trials 1, 2 and 3, respectively. Corresponding LoA values for actual and predicted VO(2max) (from RPE 9-15) were 5.4+/-11.3, 4.4+/-8.7 and 2.3+/-8.4 ml.kg(-1).min(-1), respectively. The data suggest that a sub-maximal, perceptually-guided, graded exercise protocol can provide acceptable estimates of maximal aerobic power, which are further improved with practice in fit young males. PMID- 15815938 TI - Kinematic and electromyography analysis of submaximal differences running on a firm surface compared with soft, dry sand. AB - Kinematic and electromyography (EMG) aspects of running on a firm surface and on soft, dry sand were studied to elucidate mechanisms contributing to the higher energy cost (EC) of sand running. Eight well-trained males (mean VO(2max) 64.3+/ 8.6 ml.kg(-1).min(-1)) performed barefoot running trials on a firm surface (wooden floor) and on a soft, dry sand surface (track dimensions 8.8 mx60 cm; depth 13 cm) at 8 and 11 km.h(-1). Kinematic and EMG data were collected simultaneously using an integrated six-camera 50 Hz VICON motion analysis system, an AMTI force-plate and a 10-channel EMG system. Running at 8 km.h(-1) on sand resulted in a greater (P<0.05) stance time (t(s)) compared with the firm surface. At 11 km.h(-1), sand running resulted in a greater stance-to-stride ratio (P<0.005), a shorter stride length (SL) (P<0.05), and a greater cadence (P<0.001) compared with the firm surface values. Hip and knee flexion at initial foot contact (IFC), mid-support (MS) and flexion maximum were greater (P<0.001) running on sand compared with firm surface values at 8 and 11 km.h(-1). Over duration of stride, Hamstring (semimembranosus and biceps femoris) EMG was greater running on sand compared with the firm surface at 8 (P<0.001) and 11 (P<0.05) km.h(-1). During the stance phase in the 8-km.h(-1) trials, EMG in the Hamstrings (P<0.001), Vastii (Vastus lateralis and Vastus Medialis) (P<0.02), Rectus femoris (Rec Fem) (P<0.01) and Tensor Fascia Latae (Tfl) (P<0.0001) were greater than the firm surface measures. During stance in the 11-km.h(-1) trials, Tfl EMG was greater (P<0.02) running on sand compared with the firm surface. At IFC and MS, Hamstrings' EMG was greater on sand at both running speeds (P<0.001). For the Vastii (P<0.02), Rec Fem (P<0.0001) and Tfl (P<0.0001) muscles, the EMG at MS running on sand at both speeds was greater than the firm surface values. The increased EC of running on sand can be attributed in part to the increased EMG activation associated with greater hip and knee range of motion compared with firm surface running. PMID- 15815939 TI - Intermittent short-term graded running performance in middle-distance runners in hypobaric hypoxia. AB - This study investigated whether in trained middle-distance runners, intermittent short-term graded running performance is affected by a hypobaric hypoxic environment (simulated 2,500 m) (H). Seven male middle-distance runners performed an aerobic performance test and an intermittent short-term graded anaerobic running-performance test (MART) both in H and in a normobaric normoxic environment (N). VO(2max) and OBLA were markedly lower (by 18.1% and 8.7%, respectively) in H than in N. In MART, neither maximal running velocity (V(max)) nor exhaustion-time was different between N and H (454 (7) m min(-1) vs. 451 (6) m min(-1), respectively, and 208.7 (5.2) s vs. 205.7 (4.2) s, respectively). The blood lactate concentration at sub-maximal running speed (425 m min(-1)) was significantly greater in H than in N (paired t-test: P<0.05). These results suggest that, in trained middle-distance runners, intermittent short-term graded running performance is not affected by H, despite a considerable decrease in aerobic power in H during the aerobic performance test. PMID- 15815940 TI - Nonlinear regulation of capillary perfusion in relation to ambient pO(2) changes in skeletal muscle. AB - To study the process of O(2) transport to tissue, we investigated how capillary perfusion is controlled in response to changes in tissue O(2) levels in skeletal muscle. Capillary red blood cell (RBC) velocity and perfused capillary recruitment were measured in rabbit tenuissimus muscle at various ambient oxygen tensions (pO(2)) by intravital microscopy. Both RBC velocity and capillary recruitment significantly decreased as the pO(2) level of the suffusate was increased, and the relationship between capillary perfusion, calculated from the velocity and recruitment data, and the pO(2) level of the suffusate clearly yielded a nonlinear correlation that fitted a sigmoidal curve. Capillary perfusion dramatically decreases or increases above or below a suffusate pO(2) level of around 40 Torr, where the O(2) dissociation curve of hemoglobin changes slope. These findings support the hypothesis that microvasculature possesses an intrinsic, effective flow-control mechanism by sensing the metabolic demands of tissue, intimately related to the O(2) saturation of hemoglobin. PMID- 15815941 TI - The energy cost of household and garden activities in 55- to 65-year-old males. AB - This study measured the energy expenditure of four self-paced household and garden tasks to determine whether 55- to 65-year-old men performed them at a moderate intensity [3-6 metabolic equivalents (METs)] and to predict the activity intensity via indirect methods. Resting metabolic rate and oxygen consumption were measured using Douglas bags in 50 men (X +/- SD: 60.6 +/-3.2 years, 175.8 +/ 5.6 cm, 82.6 +/- 10.1 kg ) who performed self-perceived moderate paced walking and self-paced sweeping, window cleaning, vacuuming and lawn mowing. Heart rate, CSA accelerometer counts (hip and arm), Quetelet's index, Borg rating of perceived exertion and respiratory frequency were measured as possible predictors of energy expenditure. Each of the four household and garden activities was performed at a mean intensity of > or = 3.0 METs in both the standardised laboratory environment (sweeping = 3.4, window cleaning = 3.8, vacuuming = 3.0 and lawn mowing = 5.3 METs) and the subjects' homes (sweeping = 4.1, window cleaning = 3.5, vacuuming = 3.6 and lawn mowing = 5.0 METs). Comparisons between the two settings were significantly different (p < or =0.008). Except for window cleaning, the MET values were not different from those of our previous younger sample (35-45 years). Regression analysis yielded prediction equations with 95% confidence intervals of +/-0.8 METs for both the laboratory and home environments. Although the energy expenditure means for these activities indicate that they can contribute to the 30 min day(-1) of moderate intensity physical activity required to confer health benefits, there was substantial inter individual variability. While the regression equations lack predictive precision at the individual level, they were able to determine whether energy expenditure was above the 3.0 MET threshold with correct classification rates of 91% and 94% in the laboratory and home, respectively. PMID- 15815942 TI - Meigs' syndrome with highly elevated ca. 125 levels in a patient with Takayasu arteritis: a case report. AB - A 56-year-old post-menopausal woman presented with abdominal swelling and shortness of breath. An evaluation of abdomen and pelvis showed ascites and a solid mass. Serum CA-125 level was elevated (509 IU/ml). A computerized tomography-angiography was performed since the pulses of upper extremities could not be palpated. The patient was diagnosed to be suffering from Takayasu arteritis. The exploratory laparotomy revealed a fibroma of the ovary. PMID- 15815944 TI - [When the virtuosos bump into their art]. PMID- 15815943 TI - Nephrolithiasis and nephrocalcinosis in rats with small bowel resection. AB - Intestinal resection (IR) may lead to hyperoxaluria and nephrolithiasis. A rat model of IR was developed, in which kidney stones form. We describe the urine chemistries and histopathologic features. Rats underwent resection of 40-45 cm of distal ileum (n=16) or sham resection (SR) (n=8), and were then fed a 1% Na oxalate, 0.02% Ca diet. After 1 week on the diet, 24 h urine samples were obtained for stone chemistries. At 4-7 months after surgery, kidneys were examined grossly and by light microscopy. The extent and location of crystallization was assessed by polarized light. Histochemistry and infrared spectroscopy were used to determine crystal composition. IR rats had higher urine oxalate excretion (P<0.01) and concentration (P<0.001) than SR rats, and lower urine citrate excretion; only IR rats formed kidney stones (12/15 surviving rats). Tissue calcification was found only in kidneys from IR rats, located in the cortex (83% of kidneys), medulla (73%) and papillary tip (47%). Crystals, composed of CaOx, apatite, and calcium carbonate, filled collecting duct lumens, and were associated with tubular obstruction, and interstitial inflammation. Crystals in the papillary interstitium incited inflammation with tubular destruction and development of progressive papillary erosion. This new rat model of nephrolithiasis and nephrocalcinosis resembles the pattern of urinary abnormalities and tissue calcification that may be seen in humans with small bowel resection. The model allows further studies of the mechanisms of renal crystal formation, and possible therapeutic interventions. PMID- 15815945 TI - [Clinical analysis of musicians' focal hand dystonia. Review of 86 cases]. AB - INTRODUCTION: Focal hand dystonia in musicians due to its rareness and specificity. It has been insufficiently described thus remaining a largely unknown condition. OBJECTIVE: To describe the clinical characteristics of musician's focal dystonia. METHODS AND RESULTS: We analyzed 658 musicians' cases seen during the past 4 years in a medical center for performing artists. Of the musicians treated, 86 (13 %) suffered from focal dystonia, 42 % were guitarists, 21% pianists and 6% violinists. Men were more affected than women (7:1). Sufferers reported longer practice times (4.8 hours per day) and were older (33.6 years). In comparison, other pathologies were seen when musicians were younger (26.5 years) and played no more than 3.5 hours per day. CONCLUSIONS: Focal dystonia in musicians appear to be the consequence of a long history of intense and repetitive manual work while playing music added to other factors, as for example, gender. PMID- 15815946 TI - Growing frequency of neurocysticercosis in Madrid (Spain). AB - The aim of this study is to report on the growing frequency of neurocysticercosis (NCC) in Madrid (Spain) related to changes in migratory flows. We investigated the origin of 20 patients with NCC seen over a period of 13-years (1990- 2002). Patients age and disease activity were compared with a second group of 13 patients observed in the same hospital in 1980-1989. Eighty per cent of the patients observed in the first group were foreign immigrants, all but one from South-American countries. They were young (mean age: 31.4 +- 7.7 years) and most had evidence of active disease. In contrast, only 15 % of the 13 patients observed during 1980-1989 were foreign immigrants. Recently seen Spanish-born patients were elderly (mean age: 67.3 +- 2 years), had emigrated many years earlier from rural areas where the disease was formerly prevalent, and had inactive forms of the disease. We conclude that the recent increase in the frequency of NCC in Madrid reflects a rapid growing of immigrant population from countries where the disorder is highly prevalent. Transmission of the parasite from asymptomatic T. solium carriers represents a challenge to disease prevention. Our observations should alert other European countries that may become potential destinations for similar inbound migratory flows. PMID- 15815947 TI - [Economic evaluation of acute migraine attack treatment with triptans in Spain]. AB - OBJECTIVE: To perform an economic evaluation of migraine attack treatment comparing standard doses of existing triptans in 2003, and using different outcome measurements of anti-migraine effectiveness. METHODS: A cost effectiveness analysis was performed from the National Health System perspective, using 2003 prices, comparing almotriptan 12.5 mg, eletriptan 40 mg, naratriptan 2.5 mg, rizatriptan 10 mg, sumatriptan 50 mg, sumatriptan 100 mg, zolmitriptan 2.5 mg and zolmitriptan 5 mg. Effectiveness measurements were obtained from an efficacy meta-analysis of published clinical trials, and they consisted of therapeutic gain (crude effect of triptan after placebo effect subtraction) for 2 h--anti-migraine response, pain free at 2 h, and 24 h--sustained pain free. Rescue medication use and 24 h-attack relapse rates were assessed. RESULTS: Thirty-eight clinical trials (19,872 patients) were used to assess triptans effectiveness. Eletriptan 40 mg and rizatriptan 10 mg showed the highest 24 h sustained pain free response (20.2 % in both cases), pain-free at 2 h (27.7 % and 32.2 %) and antimigraine response at 2 h (38.6 % and 31.3 %), respectively. Less rescue medication was used with eletriptan 40 mg and sumatriptan 50 mg (21 % and 20 %), and the lowest 24 h-relapse rates were observed with eletriptan 40 mg and naratriptan 2.5 mg (27 % and 21 %). Eletriptan 40 mg and sumatriptan 50 mg showed the lowest costs per successfully treated attacks with 2 h--anti-migraine response (16.50 and 17.44e) and with 24 h--sustained pain free (31.47 and 33.61e), while the lowest costs per attack that was pain free at 2 h were observed with rizatriptan 10 mg (21.36e) and eletriptan 40 mg (22.99e). CONCLUSIONS: Considering the cost-effectiveness measurements assessed, eletriptan 40 mg was the most costeffective triptan in the majority of economic analyses carried out. PMID- 15815948 TI - [Posttraumatic headache. Pathophysiology, clinical, diagnostic and therapeutic aspects]. AB - Posttraumatic headache (PTHA) is, usually, one of several symptoms of the posttraumatic syndrome and therefore may be accompanied by somatic, psychological or cognitive disturbances. The aetiology of these symptoms in individuals with mild traumatic brain injury or whiplash injury has been a subject of some controversy with explanations ranging from neural damage to malingering. PTHA can resemble a tension-type headache, migrainous or cervicogenic headaches. Post whiplash headache habitually is a pain radiating from the neck to the forehead, with moderate intensity and benign, but prolonged course. The pathogenesis of PTHA is still not well-known but might share some common headache pathways with primary headaches. In this chapter, we review recent investigations in the pathophysiology of PTHA, review recognised risk factors for a poor outcome and give some recommendations of management. We also discuss the new diagnostic criteria of IHS Classification, 2004, for PTHA and Headache attributed to whiplash. PMID- 15815949 TI - [Corticospinal tract hyperintensity: an initial finding in ALS]. PMID- 15815950 TI - [Costello syndrome. Presentation of a case with a follow-up of 35 years]. AB - OBJECTIVE: To present a patient of 35 years with Costello syndrome. MATERIAL AND METHODS: A male consulted at 11 months of age because of psychomotor delay and peculiar face. RESULTS: The main clinic features corresponded to the cutis laxa disease, although the picture and the results of the performed studies were not similar to any of the dysmorfic recognized syndromes. CONCLUSION: The follow-up of the patient during 35 years showed a long survival, despite the cardiopathy, although he has a severe psychic retardation, showed very low height and weight centiles, and very high head circumference centiles. PMID- 15815951 TI - [Thrombolytic treatment in stroke in patients with intracardiac thrombus: presentation of one case]. AB - Intravenous tissue plasminogen activator (t-PA) is a recent proven effective treatment for acute ischemic stroke patients. However, the use of t-PA in some special settings is controversial. One of these is the presence of a cardiac thrombus, given that the use of t-PA could potentially accelerate breakup of the thrombus and cause additional embolisms. The authors describe the case of a cardiological patient with a cardiac thrombus who was given IV t-PA for acute stroke treatment without complications. We discuss the necessity or not of a 24 hours delay before anticoagulants administration in these special patients. PMID- 15815952 TI - [Cavernous sinus syndrome: an initial expression of a breast carcinoma]. AB - Cavernous sinus syndrome (CSS) may be caused by a wide range of pathological process among we include metastatic disease. A case of a woman whose breast neoplasm was initially expressed by a CSS is discussed in this article. We describe a 64-year-old woman with several weeks progressive course of pain and numbness in her left-side facies and occasional diplopia. Neurological examination showed left ptosis, limitation of elevation of left eye and abducens nerve palsy as well as hypoesthesia in the frontal and maxillary regions of her left-side facies. Brain magnetic resonance imaging (MRI) scan revealed the presence of a mass in the orbit and left cavernous sinus. General examination disclosed a sclerotic and retractile lesion in her left breast which involved the nipple suggesting breast carcinoma metastases. The patient underwent a breast fine-needle aspiration biopsy which established the presence of an infiltrating breast carcinoma. This case shows the importance role of the general physical examination to determine the etiology of the CSS. It is necessary and fundamental to perform it on each patient not only to orientate further investigations but also to avoid more invasive diagnostic procedures. PMID- 15815953 TI - [Classification of epilepsies: a chaos invitation]. PMID- 15815954 TI - [Neurology in the court of the Learned King]. AB - During the reign of Alfonso X of Castile one of the most flourishing cultural periods of European history was reached. The so-called Learned King contributed to this development by promoting the use of Castilian as the language of scientific discovery instead of Latin, and favoring the recovery of Arabic origin manuscripts in the Toledo Translators' School. Where the wise men from the "three cultures" (Christian, Islamic and Jewish) worked together in harmony to perpetuate classical knowledge. The aim of this paper is to analyze neurological pathology in the period concerned. The Learned King suffered from several neurologic syndromes including delirium and coma. He also showed chronic maxillary sinusitis with orbital involvement, trigeminal neuralgia and visual loss. Furthermore, in the "Cantigas" he described miraculous treatments of several disorders including seizures, leprous neuropathy, dementia, rabies, and ergotism. PMID- 15815956 TI - Supportive oncology-palliative care: what's in a name? PMID- 15815957 TI - Integration of palliative medicine into comprehensive cancer care. AB - Because of the advent of disease-modifying agents for patients with malignancies, cancer is now a chronic illness. However, most cancer patients will experience significant symptoms and complications during the course of their illness or its treatment. In addition to their physical symptoms, patient and families are burdened with psychological, social, and spiritual difficulties. Palliative medicine addresses all these issues and complements attempts to cure the disease; it is an essential part of modern comprehensive cancer care. PMID- 15815958 TI - Palliative information: doctor-patient communication. AB - Communication between doctors and patients allows the formation of a solid working relationship based on the recognition of mutual needs and expectations. It facilitates coping by providing the right amount of diagnostic and prognostic information that assists patients and families in making therapeutic choices consistent with their life goals. In order to provide quality care, the oncologist must take an active role in encouraging disclosure of concerns and formulate a treatment plan that addresses psychosocial needs. This accomplishes the dual purpose of knowing the patient and making the patient feel known and connected to his/her professional caregivers. PMID- 15815959 TI - Palliative chemotherapy: historical perspective, applications, and controversies. AB - Despite advances in cancer detection and treatment, cancer continues to be a major public health burden in the United States, and patients with advanced or refractory cancers carry much of this burden. The primary goal of cancer treatment is cure. However, most patients with advanced, metastatic, or recurrent disease do not benefit from this intent. Recent research studies have documented the role of chemotherapy in providing symptom control, preventing complications, prolonging life, and improving quality of life (QOL) in patients with incurable cancers. Although chemotherapy under these conditions is palliative, patients receiving chemotherapy or participating in research trials are excluded from receiving much needed palliative services, such as Hospice, based on current definitions, limitations, and models of palliative care. Application of palliative services on a continuum from the time of diagnosis through the end of life (EOL) has been recognized as beneficial in the treatment of patients with terminal diseases and has been addressed through trials assessing mixed management models, providing palliative and therapeutic options. PMID- 15815960 TI - Palliative radiation therapy. AB - Radiation is an effective modality to aid in symptom management of patients with metastatic disease. The type and duration of treatment depends on the Karnofsky performance status (KPS) of the patient and type and status of the cancer. Abbreviated treatment regimens may be favored in this patient population. They provide quick palliation without the patient and family spending significant time traveling back and forth to the treatment center. Hypofractionated regimens have been found effective in relieving pain from metastatic bone disease, relieving obstruction from locally advanced lung cancer, bleeding from gynecologic cancers, and hematuria from advanced bladder cancer. More aggressive regimens such as whole-brain radiation therapy (WBRT) and stereotactic radiosurgery may be appropriate for select patients with a good KPS. Radiation has also been found to be effective in palliating recurrent cancer that has already received definitive radiation. PMID- 15815961 TI - Surgical palliative care. AB - Surgery has always played a pivotal role in care of the patient with cancer, independent of treatment intent. Recent advances have expanded that role, not only in terms of modalities available, but more broadly in terms of the expectations of the surgeon as physician involved in the interdisciplinary care of the patient with symptomatic, incurable disease. PMID- 15815962 TI - The expanding role of interventional radiology in the supportive care of the oncology patient: from diagnosis to therapy. AB - Interventional radiology (IR) plays an increasing role in the supportive care of the cancer patient from the placement of catheters to treating the offending tumor via image-guided methods. These methods entail the use of both local and regional means, with thermal ablative technology comprising the former and intra arterial embolization with radioactive particles the latter. Direct placement of the radiofrequency or cryotherapy probes into tumors that are accessible using computed tomography (CT), ultrasound, or magnetic resonance imaging (MRI) guidance provides reliable symptomatic relief of single or multiple tumors in liver, kidney, lung, bone, or soft tissue. Combined with systemic chemotherapy, these techniques increase patient survival. Intra-arterial delivery of radioactive particles to unresectable tumors both provides symptomatic relief and extends survival when combined with current systemic chemotherapy. These new approaches allow IR to provide crucial therapy for cancer patients. PMID- 15815963 TI - Palliative gastroenterology. AB - Continuous improvements in endoscopic imaging and accessories have opened up a field of interventional endoscopy. This highly technical offshoot of gastroenterology uses not just standard endoscopic techniques but also newer endoscopic ultrasound (EUS) imaging or fluoroscopic monitoring to facilitate procedures that were once performed either surgically or percutaneously, if at all. This review will update the role of these novel procedures that can be used to assist in the palliative care of patients whose malignancies involve the gastrointestinal tract. The emphasis will be on those palliative interventions that are used to overcome intestinal obstruction in the gastrointestinal tract and restore luminal patency. The role of EUS-guided celiac plexus neurolysis to assist in pain control, especially in patients with pancreatic malignancies, will also be detailed. PMID- 15815964 TI - Management of cancer pain. AB - There are four basic approaches to cancer pain control: modify the source of pain, alter central perception of pain, modulate transmission of pain to the central nervous system, and block transmission of pain to the central nervous system. Systemic pharmacologic management aimed at the first three of these approaches is the cornerstone of the treatment of most cancer patients with moderate to severe pain. Optimal pharmacologic management of cancer pain requires selection of the appropriate analgesic drug; prescription of the appropriate dose; administration of the analgesic by the appropriate route; scheduling of the appropriate dosing interval; prevention of persistent pain and relief of breakthrough pain; aggressive titration of the dose of the analgesic; prevention, anticipation, and management of analgesic side effects; consideration of sequential trials of opioid analgesics; and use of appropriate co-analgesic drugs for specific pain syndromes. Most clinicians should be able to control most of the pain in most of their cancer patients. Collaboration with pain and hospice/palliative care experts should help the rest. No cancer patient should live or die with unrelieved pain. PMID- 15815965 TI - Interventional cancer pain therapies. AB - Interventional pain therapies play a critical role in palliation of severe cancer pain. Anesthesiologists specializing in cancer pain management have developed minimally invasive techniques to: (1) optimize pain control; (2) minimize side effects, adverse outcomes, and costs; (3) enhance functional abilities and physical and psychological well-being; and (4) enhance the quality of life for cancer patients. A thorough understanding of the pathophysiology of the cancer pain is needed to implement interventional therapies. It is also important to understand the prognosis of the patient, associated comorbidities, and expectations of the patient and family. Interventional pain therapies are minimally invasive techniques that can be divided into direct drug delivery, neuroablation and neural blockade, and neurostimulation. PMID- 15815966 TI - Control of common, non-pain cancer symptoms. AB - Despite major advances in cancer biology and therapeutics, cancer and its treatment continue to cause devastating suffering. Patients with advanced cancer most often experience multiple physical and psychological symptom concurrently. We review here some of the common non-pain cancer symptoms, focusing on the assessment and treatment of fatigue, anorexia and cachexia, dyspnea, and symptoms common near the end of life. PMID- 15815967 TI - Psychosocial oncology: supportive care for the cancer patient. AB - Increasing attention is being paid to the emotional and psychosocial needs of cancer patients. As a result of huge advances in early detection and in treatment modalities, there now are millions of cancer survivors in the United States. There has been a realization that cancer survivors have distinct psychosocial needs. As cancer survivors live longer, reduction of psychological distress has been recognized as being an important part of having an improved quality of life. There have been numerous changes in the field of psychosocial oncology since it first began 25 years ago. Guidelines now exist for the definition of distress and decision trees are available for making the appropriate referrals. Advances in pharmacologic treatment for depression and anxiety have made it possible to decrease distress and increase coping in cancer patients undergoing treatment as well as in cancer survivors. Numerous individual and group therapies have demonstrated effectiveness in improving mood and quality of life in cancer patients and those at high risk for developing cancer. Due to the forthright efforts of cancer patients, there are now many organizations and list serves (e mailing lists) that cancer survivors can turn to for help before, during, and after cancer treatment. Finally, with the rapid expansion of the internet not only are there websites available as resources, but also the creation of interactive online support is becoming a reality. One of the most important issues in providing supportive care to cancer patients in the future is to meet the individual needs of patients and provide the type of psychological therapy that will work best for them. PMID- 15815968 TI - Cancer rehabilitation. AB - Cancer-related functional impairments and the associated economic burden are likely to increase in coming decades. A variety of factors contribute to the current availability of comprehensive rehabilitation services in only a few cancer centers. In general, cancer patients are highly distressed by the loss of independent mobility and self-care. The majority of patients are amenable to receiving rehabilitation services. Reports describing the integration of rehabilitation specialists into the delivery of cancer care strongly suggest that patients independence can be optimized through established techniques. The provision of humane and effective rehabilitation requires the formulation of dynamic and clinically appropriate goals. Many goals are common to most malignancies, eg, reducing the functional impact of aerobic deconditioning and chemotherapeutic neuropathy. Others are highly disease-specific, such as scapular stabilization following cranial nerve IX sacrifice. The expectations of patients and clinicians must evolve in response to the progression of disease. Restorative, supportive, preventative, and palliative goals should be re evaluated at critical points along the disease course. A rubric for the development and implementation of rehabilitation goals at different cancer stages is described and illustrated with examples from breast and head and neck cancers. PMID- 15815969 TI - Complementary medicine in chronic cancer care. AB - Although advancements in cancer care have led to increased cure rates and survival times, those coping with the long-term sequelae of a cancer diagnosis often report high levels of distress, poor health-related quality of life, and unmet psychosocial needs. The shortcomings of the conventional biomedical system at sufficiently addressing these chronic illness issues are a primary reason that many patients turn to complementary and alternative medicine (CAM). Although patients usually use such treatments in addition to conventional care, they often do not discuss doing so with their conventional physicians, who are perceived as being unreceptive to the topic. Physicians may feel uninformed about CAM treatments, especially given the relatively small amount of data on the topic available in the medical literature. This article reviews a few CAM modalities commonly used by cancer survivors and those with chronic cancer illness, with a particular focus on familiarizing physicians with treatments that may potentially complement regular oncologic care by decreasing stress and enhancing health related quality of life. PMID- 15815970 TI - Spirituality and healing. AB - Spirituality can exert a tremendous impact on ones health and promote recovery from trauma and illness, including cancer. Throughout the history of mankind, spirituality and religion have played a major role in healing a variety of physical and mental illnesses. Cancer is one of the most devastating illnesses, as it affects ones physical, emotional, psychological, and spiritual well-being. An increasing body of scientific literature supports the concept that spirituality can significantly improve healing from cancer and promote the coping response of caregivers and healthcare professionals. We believe that spirituality is an important component of the healing process and should be integrated with conventional medicine to treat this complex disease. PMID- 15815971 TI - Sedation for the relief of refractory symptoms in the imminently dying: a fine intentional line. AB - There is a continuum of the goals of comfort and function in palliative care that begins with comfort and function being equal priorities and sedation being unacceptable. As disease progresses, the goals and preferences of the patient turn to coping with the loss of function caused by the disease and acceptance of unintentional sedation from the disease, its therapies, or symptom relief interventions. As patients approach the end of life, they may need intentional sedation for the relief of refractory symptoms. Such sedation can be divided into three categories: routine, infrequent, and extraordinary with respect to the frequency, difficulty, and risks involved with the drugs and routes of administration required to induce and maintain a level of sedation that relieves the patient's physical and existential symptoms. Extraordinary sedation with continuous infusions of midazolam, thiopental, and propofol can relieve refractory symptoms in most patients in their final days of life. Palliative care clinicians should become comfortable with the ethical justification and technical expertise needed to provide this essential, extraordinary care to the small but deserving number of patients in whom routine and infrequent sedation does not adequately relieve their suffering. PMID- 15815973 TI - Comparative molecular dynamics simulations of HIV-1 integrase and the T66I/M154I mutant: binding modes and drug resistance to a diketo acid inhibitor. AB - HIV-1 IN is an essential enzyme for viral replication and an interesting target for the design of new pharmaceuticals for use in multidrug therapy of AIDS. L 731,988 is one of the most active molecules of the class of beta-diketo acids. Individual and combined mutations of HIV-1 IN at residues T66, S153, and M154 confer important degrees of resistance to one or more inhibitors belonging to this class. In an effort to understand the molecular mechanism of the resistance of T66I/M154I IN to the inhibitor L-731,988 and its specific binding modes, we have carried out docking studies, explicit solvent MD simulations, and binding free energy calculations. The inhibitor was docked against different protein conformations chosen from prior MD trajectories, resulting in 2 major orientations within the active site. MD simulations have been carried out for the T66I/M154I DM IN, DM IN in complex with L-731,988 in 2 different orientations, and 1QS4 IN in complex with L-731,988. The results of these simulations show a similar dynamical behavior between T66I/M154I IN alone and in complex with L 731,988, while significant differences are observed in the mobility of the IN catalytic loop (residues 138-149). Water molecules bridging the inhibitor to residues from the active site have been identified, and residue Gln62 has been found to play an important role in the interactions between the inhibitor and the protein. This work provides information about the binding modes of L-731,988, as well as insight into the mechanism of inhibitor-resistance in HIV-1 integrase. PMID- 15815974 TI - The CCPN data model for NMR spectroscopy: development of a software pipeline. AB - To address data management and data exchange problems in the nuclear magnetic resonance (NMR) community, the Collaborative Computing Project for the NMR community (CCPN) created a "Data Model" that describes all the different types of information needed in an NMR structural study, from molecular structure and NMR parameters to coordinates. This paper describes the development of a set of software applications that use the Data Model and its associated libraries, thus validating the approach. These applications are freely available and provide a pipeline for high-throughput analysis of NMR data. Three programs work directly with the Data Model: CcpNmr Analysis, an entirely new analysis and interactive display program, the CcpNmr FormatConverter, which allows transfer of data from programs commonly used in NMR to and from the Data Model, and the CLOUDS software for automated structure calculation and assignment (Carnegie Mellon University), which was rewritten to interact directly with the Data Model. The ARIA 2.0 software for structure calculation (Institut Pasteur) and the QUEEN program for validation of restraints (University of Nijmegen) were extended to provide conversion of their data to the Data Model. During these developments the Data Model has been thoroughly tested and used, demonstrating that applications can successfully exchange data via the Data Model. The software architecture developed by CCPN is now ready for new developments, such as integration with additional software applications and extensions of the Data Model into other areas of research. PMID- 15815975 TI - Helix-coil transition of alanine peptides in water: force field dependence on the folded and unfolded structures. AB - The force fields used in classical modeling studies are semiempirical in nature and rely on their validation by comparison of simulations with experimental data. The all-atom replica-exchange molecular dynamics (REMD) methodology allows us to calculate the thermodynamics of folding/unfolding of peptides and small proteins, and provides a way of evaluating the reliability of force fields. We apply the REMD to obtain equilibrium folding/unfolding thermodynamics of a 21-residue peptide containing only alanine residues in explicit aqueous solution. The thermodynamics of this peptide is modeled with both the OPLS/AA/L and the A94/MOD force fields. We find that the helical content and the values for the helix propagation and nucleation parameters for this alanine peptide are consistent with measurements on similar peptides and with calculations using the modified AMBER force field (A94/MOD). The nature of conformations, both folded and unfolded, that contributes to the helix-coil transition profile, however, is quite different between these two force fields. PMID- 15815977 TI - Interaction and structural study of kinin peptide bradykinin and ganglioside monosialylated 1 micelle. AB - Partitioning of small proteins and peptides from the aqueous to membrane phase is often coupled with folding. In this work we examine the binding and folding of the kinin peptide, bradykinin (BK), in the presence of the ganglioside monosialylated 1 (GM1) micelle. Using two-dimensional NMR techniques, we have shown that at low concentration, GM1 micelle is able to induce a turn conformation to BK. A pulsed-field gradient diffusion NMR study indicated that the peptide partitions into the GM1 micelle with a DeltaG(part) of -3.14 +/- 0.03 kcal/mol. A saturation transfer difference (STD) NMR study indicated that the binding is mostly through hydrophobic residues. PMID- 15815978 TI - Structural characterization of an iron-sulfur cluster assembly protein IscU in a zinc-bound form. PMID- 15815984 TI - Quantitative analysis of helix-coil transition of block copolypeptide, Glu12 Ala12, by combined use of CD and NMR spectroscopy. AB - To investigate helix-coil transition mechanisms, conformations of Glu12-Ala12, EA, in aqueous solution have been studied in detail over the pH range from 2 to 8 and the temperature range from 20 to 60 degrees C using CD and NMR spectroscopy. The 750-MHz NMR spectra displayed excellent dispersion of the backbone amide proton signals, and permitted essentially complete sequence-specific resonance assignments. These assignments, together with short- and medium-range nuclear Overhauser effect (NOE) constraints and coupling constants, enable us to analyze conformational characteristics of all the residues in the EA peptide individually. A combined use of CD and NMR techniques reveals that the EA peptide assumes a stable alpha-helix from Glu12 to Ala19 in 0.1 M NaCl solution at 20 degrees C above pH 7. The alpha-helix is getting longer as decreasing pH. Below pH 4, the peptide assumes the longest alpha-helix from Glu3 to Ala23. The important observation of the present study is that the helix-coil transition occurs stepwise, residue by residue, from both the N- and C-termini of the alpha helix. No conformational equilibrium between the helical and random-coil states is detected for the residues in the central region of the alpha-helix. Quantitative analysis of temperature-induced helix-to-coil transitions at various pHs provides a pH-independent residual enthalpy change delta H(r) = 0.95 kcal res(-1). Similar values have been reported for a 50-residue alanine-rich peptide (1.2 kcal res(-1)), poly-L-glutamate (1.1 kcal res(-1)), poly-L-lysine (1.1 kcal res(-1)), and poly-L-alanine (0.86 kcal res(-1)). Those investigations, along with our present result, suggest that delta H(r) is mainly determined by the transformation of the backbone associated with the disruption of the intramolecular hydrogen bond. These results should increase our understanding of the helix-coil transition. PMID- 15815985 TI - A well-defined amphipathic conformation for the calcium-free cyclic lipopeptide antibiotic, daptomycin, in aqueous solution. AB - Daptomycin is a 13-residue cyclic lipopeptide with Ca2+-dependent bactericidal activity against a variety of high-risk pathogens. Ring closure in daptomycin is via an ester linkage between the side chain of Thr4 and the C-terminal carboxyl of the main chain; the N-terminal residue is capped by a decanoyl aliphatic chain. Extensive NMR data obtained under solution conditions that minimize aggregation have provided constraints for a detailed conformational analysis of daptomycin in aqueous solution, which should facilitate the rational design of improved analogs and enhance understanding of its mode of action. Transannular and shorter-range nuclear Overhauser effects (NOEs) as well as amide temperature shifts and 3J(NH alpha) coupling constants indicate that daptomycin adopts a well defined conformation containing a distorted hairpin formed by Gly5-D-Ala6 type II' beta-turn. A number of hydrophobic moieties (the lipid N-cap and the Trp1 and Kyn13 side chains) are clustered at one end of the hairpin, while neutral polar and anionic residues are localized on the other end, leading to amphipathicity in the molecule. These features suggest a mode of action in which the large hydrophobic cluster of the peptide interacts with the acyl chain region of a membrane. This interaction may be facilitated by Ca2+ ions, both by neutralizing the anionic charges and by favoring association with the membrane head groups. Interestingly, our findings differ from two recent articles in which the aqueous conformation of Ca2+-free daptomycin is reported to lack a well-defined conformation (D. Jung, A. Rozek, M. Okron, and R. E. W. Hancock, Chemistry & Biology, 2004, Vol. 11, pp. 949-957) or is suggested to populate an alternate conformation (L.-J. Ball, C. M. Goult, J. A. Donarski, J. Micklefield, and V. Ramesh, Organic & Biomolecular Chemistry, 2004, Vol. 2, pp. 1872-1878). PMID- 15815986 TI - Proteome analysis of Arabidopsis thaliana by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. AB - In the present study we show results of a large-scale proteome analysis of the recently sequenced plant Arabidopsis thaliana. On the basis of a previously published sequential protein extraction protocol, we prepared protein extracts from eight different A. thaliana tissues (primary leaf, leaf, stem, silique, seedling, seed, root, and inflorescence) and analysed these by two-dimensional gel electrophoresis. A total of 6000 protein spots, from three of these tissues, namely primary leaf, silique and seedling, were excised and the contained proteins were analysed by matrix assisted laser desorption/ionisation time of flight mass spectrometry peptide mass fingerprinting. This resulted in the identification of the proteins contained in 2943 spots, which were found to be products of 663 different genes. In this report we present and discuss the methodological and biological results of our plant proteome analysis. PMID- 15815987 TI - Identification of shed proteins from Chinese hamster ovary cells: application of statistical confidence using human and mouse protein databases. AB - The shedding process releases ligands, receptors, and other proteins from the surface of the cell and is a mechanism whereby cells communicate. Even though altered regulation of this process has been implicated in several diseases, global approaches to evaluate shed proteins have not been developed. A goal of this study was to identify global changes in shed proteins in media taken from cells exposed to low-doses of radiation to develop a fundamental understanding of the bystander response. Chinese hamster ovary cells were chosen because they have been widely used for radiation studies and are reported to respond to radiation by releasing factors into the media that cause genomic instability and cytotoxicity in unexposed cells, i.e., a bystander effect. Media samples taken for irradiated cells were evaluated using a combination of tandem- and Fourier transform-ion cyclotron resonance (FT-ICR)-mass spectrometry (MS) analyses. Since the hamster genome has not been sequenced, MS data was searched against the mouse and human protein databases. Nearly 150 proteins identified by tandem mass spectrometry were confirmed by FT-ICR. When both types of MS data were evaluated, using a new confidence scoring tool based on discriminant analyses, about 500 proteins were identified. Approximately 20% of these identifications were either integral membrane proteins or membrane associated proteins, suggesting that they were derived from the cell surface and, hence were likely shed. However, estimates of quantitative changes, based on two independent MS approaches, did not identify any protein abundance changes attributable to the bystander effect. Results from this study demonstrate the feasibility of global evaluation of shed proteins using MS in conjunction with cross-species protein databases and that significant improvement in peptide/protein identifications is provided by the confidence scoring tool. PMID- 15815988 TI - A calix[4]imidazolium[2]pyridine as an anion receptor. PMID- 15815990 TI - Translation of STO charge distributions. AB - Barnett and Coulson's zeta-function method (M. P. Barnett and C. A. Coulson, Philos. Trans. R. Soc., Lond. A 1951, 243, 221) is one of the main sources of algorithms for the solution of multicenter integrals with Slater-type orbitals. This method is extended here from single functions to two-center charge distributions, which are expanded at a third center in terms of spherical harmonics times analytical radial factors. For s-s distributions, the radial factors are given by a series of factors corresponding to the translation of s type orbitals. For distributions with higher quantum numbers, they are obtained from those of the s-s distributions by recurrence. After analyzing the convergence of the series, a computational algorithm is proposed and its practical efficiency is tested in three-center (AB/CC) repulsion integrals. In cases of large basis sets, the procedure yields about 12 correct significant figures with a computational cost of a few microseconds per integral. PMID- 15815991 TI - Zori 1.0: a parallel quantum Monte Carlo electronic structure package. AB - The Zori 1.0 package for electronic structure computations is described. Zori performs variational and diffusion Monte Carlo computations as well as correlated wave function optimization. This article presents an overview of the implemented methods and code capabilities. PMID- 15815992 TI - Palladium(0)-catalyzed tandem cyclization of allenenes: direct construction of tricyclic heterocycles through aromatic C--H activation. AB - Palladium(0)-catalyzed tandem cyclization of allenenes is described. Treatment of allenenes with an aryl halide, potassium carbonate, and catalytic [Pd(PPh(3))(4)] in dioxane afforded tri- or tetracyclic heterocycles in moderate to good yields through insertion of arylpalladium(II) halide into the allenic moiety, intramolecular carbopalladation, and aromatic C--H bond activation. The substituent on the olefin terminus has proven to be essential for the success of the tandem cyclization. The reaction with heterocyclic aryl halides such as iodopyrazine or 4-bromo-1-methylindole afforded tri- or tetracyclic heteroaromatic products in good yields. PMID- 15815994 TI - Synthesis, spectroscopic and nonlinear optical properties of multiple [60]fullerene-oligo(p-phenylene ethynylene) hybrids. AB - A series of multiple [60]fullerene terminated oligo(p-phenylene ethynylene) (OPE) hybrid compounds has been synthesized through a newly developed in situ ethynylation method. Structural and magnetic shielding properties of the highly unsaturated carbon-rich C(60) and OPE scaffolds were characterized by 1D and 2D NMR spectroscopic analyses. Electronic interactions between the [60]fullerenes and the OPE backbones were investigated by UV/Vis spectroscopic and cyclic voltammetry (CV) experiments. Our studies clearly show that although the multiple [60]fullerene groups are connected via pi-conjugated OPE frameworks, they present diminutive electronic interactions in the ground state, and the electronic behavior of the [60]fullerene cages are only affected by the OPE backbones through modest inductive effects. Interestingly, sizable third-order nonlinear optical (NLO) responses (gamma) and enhanced two-photon absorption (TPA) cross sections (sigma((2))) were determined for the multifullerene-OPE hybrid 31 relative to its OPE precursor from differential optical Kerr effect (DOKE) experiments. Such enhanced NLO performance is presumably due to the occurrence of periconjugation and/or charge transfer effects in the excited state. In addition, comparatively strong excited-state absorption was observed and characterized for OPE pentamer 12. Thus, the use of such fullerene-derivatized conjugated oligomers aids the quest for molecules with large third-order NLO and TPA properties. PMID- 15815995 TI - Syntheses, structures, photoluminescence and theoretical studies of xanthone in crystalline resorcinarene-based inclusion complexes. AB - Two new crystalline resorcinarene-based xanthone inclusion complexes, CECRxanthoneMeOH (1), and HECR2 xanthone6 MeOH (2) (CECR = C ethylcalix[4]resorcinarene, HECR = hexaethylresorcin[6]arene) have been prepared to study the relation between photophysical properties and solid-state structure. Compared with the neat crystals, the xanthone phosphorescence is severely quenched in both solids, but the lifetime is an order of magnitude larger in 2, in which xanthone occurs as a dimer, than in 1, in which it occurs as a monomer. The electronic transitions involved in the photoluminescent process, and the relation between the energy levels of host and guest and emission quenching of the guest in the supramolecular solid have been investigated by means of time dependent density functional theory (TDDFT) calculations. PMID- 15815996 TI - Salt removal during Off-Gel electrophoresis of protein samples. AB - The Off-Gel technology was recently described for protein fractionation in a solution placed on top of an immobilized pH gradient gel. In addition, this process was found to remove salts from the biological samples to analyze. This desalting effect is studied experimentally in a conductometric prototype cell. A simplified analytical model is developed to understand this process and a good agreement is found with the conductivity measurements. To illustrate the desalting of a biological sample, a 1 mg.mL(-1) solution of beta-lactoglobulin A in 0.1 M NaCl is subjected to electrophoresis in a single compartment Off-Gel cell. The analysis of the resulting sample by ESI-MS demonstrates the effective removal of salt. A finite element diffusion-migration model is also used to illustrate how the nonuniformity of the electric field in the cell, associated with the salt migration, can slow down the desalting process. PMID- 15815997 TI - Timing of futility analyses for 'proof of concept' trials. AB - 'Proof of concept' (PoC) trials often are carried out to determine if a treatment is biologically active or inactive. If the predictive probability of rejecting a null hypothesis of no treatment effect on completion of the trial calculated using information accumulated before trial completion is sufficiently low, then the trial could be terminated and resources redirected more productively. If the predictive probability is high, some time might be gained by proceeding with further steps in drug development without affecting the course of the PoC trial in any way. We describe how the interim evaluation timing affects the ability to reach a decision to stop or continue as a function of various assumptions about the true parameters and about the interim outcomes. An important practical finding is that there is no point to carrying out an evaluation before accumulating about 40 per cent of the planned observations. Consequently, if trial costs are mostly at startup or for recruitment of subjects for extended periods of observation, it is unlikely that much benefit will be realized by considering the possibility of early termination for futility except possibly for administrative efficiencies with respect to resource and staff allocation. PMID- 15815998 TI - Proteome investigation of the global regulatory role of sigma 54 in response to gentisate induction in Pseudomonas alcaligenes NCIMB 9867. AB - Pseudomonas alcaligenes NCIMB 9867 (strain P25X) utilizes the gentisate pathway for the degradation of aromatic hydrocarbons. The gene encoding the alternative sigma (sigma) factor sigma(54), rpoN, was cloned from strain P25X and a rpoN knock-out strain, designated G54, was constructed by insertional inactivation with a kanamycin resistance gene cassette. The role of sigma(54) in the physiological response of P. alcaligenes P25X to gentisate induction was assessed by comparing the global protein expression profiles of the wild-type P25X with the rpoN mutant strain G54. Analysis of two-dimensional polyacrylamide gel electrophoresis gels showed that 39 out of 355 prominent protein spots exhibited differential expression as a result of the insertional inactivation of rpoN. Identification of the protein spots by matrix-assisted laser desorption/ionization-time of flight/time of flight revealed a wide diversity of proteins that are affected by the sigma(54) mutation, the largest group being proteins that are involved in carbon metabolism. The strictly inducible gentisate 1,2-dioxygenase, one of two isofunctional copies of the key enzyme in the gentisate pathway, and enzymes of the TCA cycle, pyruvate metabolism and gluconeogenesis were part of this group. Other proteins that are part of the sigma(54) regulon include enzymes implicated in nitrogen metabolism, transport proteins, stress-response proteins and proteins involved in cell motility. The results of this study showed that sigma(54) plays a global regulatory role in the expression of a wide variety of genes in P. alcaligenes, including the wild-type response to the presence of the aromatic inducer, gentisate. PMID- 15815999 TI - Detection of prion protein using a capillary electrophoresis-based competitive immunoassay with laser-induced fluorescence detection and cyclodextrin-aided separation. AB - The development of capillary electrophoresis (CE)-based competitive immunoassay for prion protein (PrP) using carboxymethyl beta-cyclodextrin (CM-beta-CD) as a buffer additive is described here. The assay was based on the competitive binding of PrP and a fluorescein-labeled peptide from the prion protein with a limiting amount of specific antibody. The amount of both free and fluorescein-labeled peptide bound to antibody (immunocomplex) were determined by CE with laser induced fluorescence detection. In the presence of PrP, the peak height ratio of the immunocomplex and the free peptide was altered compared to the control. These changes were directly proportional to the amount of PrP present. The fluorescently labeled peptide spanning amino acid positions 140-158 of the PrP and its corresponding monoclonal antibody is reported here. The reaction times of the antibody with either the peptide or the recombinant PrP was less than 1 min and is a large improvement over the 16-18 h required to achieve equilibrium for polyclonal antibodies. CM-beta-CD was explored as a buffer additive to suppress analyte adsorption and enhance separation selectivity in the CE analysis. A fast (1.1 min), selective (resolution 4.7), and reproducible (relative standard deviations of migration time for free and bound fluorescein isothiocyanate (FITC) peptide 0.56% and 0.64%, respectively) separation was obtained with 0.6% CM-beta CD in 25 mM N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS) at pH 8.8. The concentration detection limit of the assay for recombinant PrP was determined to be 80 ng/mL (or mass detection limit 1 pg). When blood samples from scrapie-infected sheep and from normal sheep were tested, the results of the blood assay were consistent with scrapie status of the sheep as determined post mortem by Western blot analysis. Development of this assay will lead to a potentially robust, rapid, and specific preclinical diagnosis for transmissible spongiform encephalopathies (TSEs) in animals and humans. PMID- 15816000 TI - Dual regulation of BCR-mediated growth inhibition signaling by CD72. AB - CD72 has been reported to regulate BCR-mediated signals both positively and negatively. SHP-1 and Grb2 bind, respectively, to ITIM1 and ITIM2 of CD72. We generated transformed B cell lines with an immature phenotype following J2 virus infection of splenocytes from CD72(-/-) and wild-type (Wt) mice. The transformed lines were infected with retroviral vectors carrying Tyr (Y) to Phe (F) substitutions in the ITIM sequences (ITIM1 mutated: Y7/F; ITIM2 mutated: Y39/F; and both ITIM mutated: Y7,39/F). Cross-linking of the BCR induced growth inhibition in transfectants expressing Wt CD72, but this response was less sensitive in transfectants with Y7,39/F. The Y7/F transfectants demonstrated the least sensitive response. We were not able to obtain transfectants with Y39/F, suggesting that CD72 associated with SHP-1, but not with Grb2, delivers a strong negative signal. Pre-ligation of CD72, which induces dephosphorylation of the molecule, partially rescued the Wt transfectants from growth inhibition, leading to a growth response profile similar to that of Y7,39/F transfectants. These results suggest that ITIM1/SHP-1 delivers a very strong negative signal that is down-modulated by signals through ITIM2/Grb2, leading to delivery of an attenuated negative signal. Thus, pre-ligation of CD72 results in the manifestation of an ostensible positive signal. PMID- 15816001 TI - Distinct in vivo dendritic cell activation by live versus killed Listeria monocytogenes. AB - Immunization of mice with live or heat-killed Listeria monocytogenes (HKLM) efficiently primes pathogen-specific CD8(+) T cells. T lymphocytes primed by HKLM, however, undergo attenuated proliferation and do not fully differentiate. Thus, only infection with live bacteria induces long-term, CD8(+) T cell-mediated protective immunity. In this study we demonstrate that live and heat-killed bacteria, while both associating with Mac-3(+)CD11b(hi) cells, localize to distinct splenic areas following intravenous inoculation. While HKLM localize to the marginal zone and the splenic red pulp, live L. monocytogenes are carried to the T cell zone of splenic white pulp. Despite these differences, in vivo depletion of CD11c-expressing cells prevents priming of naive T cells by either HKLM or live L. monocytogenes. Analysis of CD11c(hi) dendritic cells (DC) reveals that infection with live L. monocytogenes induces higher levels of CD40, CD80 and CD86 expression than immunization with HKLM. Our results suggest that CD8(+) T cell priming following HKLM immunization or live infection is mediated by DC and that the disparate outcomes of priming can be attributed to suboptimal conditioning of DC in the absence of live, cytosol-invasive bacteria. PMID- 15816002 TI - Limited correlation between human thymus and blood NKT cell content revealed by an ontogeny study of paired tissue samples. AB - NKT cells are a CD1d-restricted T cell subset with strong immunoregulatory properties. Human NKT deficiencies are associated with autoimmune diseases such as type 1 diabetes and several types of cancer, yet there is little understanding of how the human NKT cell pool develops or is maintained. In this study, we present the first detailed analysis of human NKT cells from donor-matched postnatal thymus and blood samples. In mice, NKT cells are a thymus-dependent population that migrates to the periphery at an immature stage. Our data show that human NKT cells also undergo early stages of development in the thymus, forming a CD4(+)CD161(-/low) population that predominates neonatal thymic and blood NKT cell pools. CD4(-) and CD161(+) NKT cells accumulate with age in the blood, but not thymus, to the point that they dominate the NKT cell compartment in adult blood. This is consistent with the post-thymic maturation of NKT cells exported from the thymus at the putatively immature CD4(+)CD161(-/low) stage. Interestingly, while thymus and peripheral NKT cell frequencies vary widely between patients and are relatively stable between age groups, there is no clear relationship between the NKT cell frequency in thymus and blood. PMID- 15816003 TI - Determination of partial amino acid composition from tandem mass spectra for use in peptide identification strategies. AB - We demonstrate a new approach to the determination of amino acid composition from tandem mass spectrometrically fragmented peptides using both experimental and simulated data. The approach has been developed to be used as a search-space filter in a protein identification pipeline with the aim of increased performance above that which could be attained by using immonium ion information. Three automated methods have been developed and tested: one based upon a simple peak traversal, in which all intense ion peaks are treated as being either a b- or y ion using a wide mass tolerance; a second which uses a much narrower tolerance and does not perform transformations of ion peaks to the complementary type; and the unique fragments method which allows for b- or y-ion type to be inferred and corroborated using a scan of the other ions present in each peptide spectrum. The combination of these methods is shown to provide a high-accuracy set of amino acid predictions using both experimental and simulated data sets. These high quality predictions, with an accuracy of over 85%, may be used to identify peptide fragments that are hard to identify using other methods. The data simulation algorithm is also shown post priori to be a good model of noiseless tandem mass spectrometric peptide data. PMID- 15816004 TI - A strategy for high-resolution protein identification in surface-enhanced laser desorption/ionization mass spectrometry: calgranulin A and chaperonin 10 as protein markers for endometrial carcinoma. AB - Surface-enhanced laser desorption/ionization-mass spectrometry (SELDI-MS) has conventionally been practiced on linear time of flight (TOF) which has low mass accuracy and resolution. Here we demonstrate in an examination of both malignant and nonmalignant endometrial tissue homogenates that high mass accuracy and resolution in the MS stage are crucial. Using a commercially available quadrupole/TOF (QqTOF), we were able to resolve two potential cancer markers, subsequently identified off-line as chaperonin 10 and calgranulin A, that differ by 8 Da in mass. Two off-line protein identification protocols were developed: the first was based on size-exclusion chromatography (SEC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), protein extraction, trypsin digestion, and matrix-assisted laser desorption/ionization-tandem MS (MALDI-MS/MS); the second on SEC and shotgun nano-liquid chromatography (nanoLC) MS/MS. Analyses on a cohort of 44 endometrial homogenates showed 22 out of 23 nonmalignant samples had nondetectable to very low abundance of chaperonin 10 and calgranulin A; 17 of the 21 malignant samples had detectable to abundant levels of both proteins. Immunohistochemical staining of a tissue microarray of 32 samples showed that approximately half of malignant endometrial tissues exhibited positive staining for calgranulin A in the malignant epithelium, while 9 out of 10 benign tissues exhibited negative epithelial staining. In addition, macrophages/granulocytes in malignant as well as nonmalignant tissues showed positive staining. No immunostaining occurred in stroma or myometrium. Calgranulin A, in combination with chaperonin 10 and other proteins, may eventually constitute a panel of markers to permit diagnosis and screening of endometrial cancer. PMID- 15816005 TI - Use of two-dimensional gel electrophoresis in predictive toxicology: identification of potential early protein biomarkers in chemically induced hepatocarcinogenesis. AB - Our current approach focused on the identification of potential early protein biomarker signatures which are indicative of the carcinogenic processes in rats exposed to 20 mg/kg of the liver carcinogen N-nitrosomorpholine (NNM). Treated liver was investigated at different timepoints. Therefore, proteins were separated by two-dimensional gel electrophoresis as a first step prior to identification of differentially expressed proteins by mass spectrometry. Proteomic analysis of liver samples after one day of exposure revealed significant upregulation of proteins involved in response to cellular stress induced by NNM (superoxide dismutase, heat shock protein 60, peroxiredoxin). Eighteen weeks after withdrawal of NNM, we were able to identify cancer-related proteins in rat liver bearing malignant, transformed cells (caspase-8 precursor, vimentin, Rho GDP dissociation inhibitor). Some of these proteins were already deregulated after three weeks of exposure indicating their potential usefulness as early predictive biomarkers for liver carcinogenicity (annexin A5, fructose 1,6-bisphosphatase). As regulatory toxicology approaches usually include the investigation of carcinogenicity in two-years studies in rodents, especially the detection of early protein biomarker signatures which precede the appearance of neoplasia, demonstrates the high potential of proteomics approaches to substantially reduce the time and costs of carcinogenicity testing. PMID- 15816006 TI - Optimised two-dimensional electrophoresis procedures for the protein characterisation of structural tissues. AB - The protein analysis of structural tissues is typically highly problematic. Amniotic membrane displays unique wound healing and anti-scarring properties; however, little is known concerning its active protein content. The structural nature of amniotic membrane necessitated development and extensive optimisation of the entire two-dimensional (2-D) workflow. Proteins were extracted using powerful solubilisation buffers and analysis carried out using 2-D electrophoresis followed by mass spectrometry (MS) identification. Preservation and processing resulted in prefractionation of soluble from structural and membrane-associated proteins. Enhanced protein solubility was achieved by cysteine blocking using both N,N-dimethylacrylamide (DMA) alkylation and bis(2 hydroxyethyl) disulphide (HED); an alternative procedure for the effective application of HED is demonstrated. The benefits of precipitation and cup-loading versus in-gel rehydration were also assessed, with procedures for the employment of HED with the latter described. Following optimisation, a representative sample 21 proteins were identified from amniotic membrane using MS verify procedures were MS-compatible. Our results demonstrate that techniques for the reproducible separation of proteins from a proteinaceous structural tissue have been optimised. Briefly, proteins are extracted using a thiourea/urea extraction buffer containing carrier ampholytes, dithiothreitol (DTT), and 3 (cyclohexylamino)-1-propanesulfonic acid (CHAPS). After DMA alkylation, proteins were precipitated (using the 2-D clean-up kit from Amersham Biosciences) and resolubilised in extraction buffer containing a lower concentration of DTT. Samples were either cup-loaded onto rehydrated HED-containing strips or rebuffered into HED-containing buffer followed by in-gel rehydration. PMID- 15816007 TI - Doppler-derived umbilical artery absolute velocities and their relationship to fetoplacental volume blood flow: a longitudinal study. AB - OBJECTIVES: To construct reference ranges for serial measurements of the umbilical artery (UA) absolute blood flow velocities in the second half of pregnancy and to test the hypothesis that significant associations exist between UA velocities and placental volume blood flow assessed from umbilical vein blood flow velocities and diameter. METHODS: This was a prospective longitudinal study of the umbilical circulation. UA absolute velocities and umbilical vein blood flow were measured at 4-weekly intervals between 19 and 42 weeks' gestation in 130 low-risk singleton pregnancies. A total of 511 observations were used to construct the reference ranges and assess the association between UA absolute velocities and placental volume flow using multilevel modeling. RESULTS: Both UA absolute velocities and placental volume blood flow showed a steady increase throughout the second half of pregnancy. However, the gestational age-related increase in the UA end-diastolic velocity (EDV) was greater than the corresponding increase in the peak systolic velocity (PSV). The time-averaged intensity-weighted mean velocity (TAWMV)/time-averaged maximum velocity (TAMXV) was 0.6 indicating probably not a completely parabolic velocity profile. There was a significant positive association (P < 0.00001) between UA absolute velocities and placental volume blood flow, but this association was modified by the gestational age. The intraobserver coefficients of variation for the UA PSV, EDV, TAMXV and TAWMV and placental volume blood flow were 10.17%, 16.29%, 11.46%, 18.18% and 8.61%, respectively. CONCLUSION: We have established new reference ranges for the UA absolute velocities based on longitudinal data. They show a significant association with fetoplacental volume blood flow and may have a clinical value in the assessment of the umbilical circulation. PMID- 15816008 TI - Assessment of the gap between the fetal nasal bones at 11 to 13 + 6 weeks of gestation by three-dimensional ultrasound. AB - OBJECTIVE: To detect the presence of a gap between the fetal nasal bones at 11 to 13 + 6 weeks of gestation and to verify if this gap could lead to the erroneous diagnosis of absent nasal bone. METHODS: Three-dimensional (3D) ultrasound was used to assess the fetal nose in 450 singleton pregnancies, immediately after two dimensional (2D) evaluation of the nasal bones and screening for chromosomal defects by a combination of maternal age and the measurement of fetal nuchal translucency at 11 to 13 + 6 (median, 12) weeks of gestation. A 3D volume of the fetal face was acquired and then analyzed using the multiplanar mode. A sequence of transverse views was used to confirm the presence or absence of the nasal bones and when they were present any visible gap between them was measured. A perfectly mid-sagittal plane was then examined to determine if the nasal bone was visible or not. RESULTS: In 421/450 (93.6%) cases the nasal bone was present during 2D ultrasound. Using the multiplanar mode of 3D ultrasound, in 83/421 (19.7%) fetuses a gap between the nasal bones could be demonstrated and in 36/83 (43.4%) cases the nasal bone was found to be absent in the perfect mid-sagittal view. In 29/450 (6.4%) cases the nasal bones were absent during the 2D scan. In the 3D assessment there was absence of both bones in 25/29 (86.2%) cases and absence of one of the two bones in 4/29 (13.8%) cases. Chorionic villus sampling demonstrated that the fetal karyotype was normal in 404 and abnormal in 46 cases, including 31 cases of trisomy 21. There was absence of one or both nasal bones in three (0.7%) of the chromosomally normal fetuses, in 19 (61.3%) with trisomy 21 and in seven (46.7%) with other chromosomal defects. CONCLUSIONS: At 11 to 13 + 6 weeks of gestation there is a gap between the nasal bones in about 20% of fetuses, and in about 40% of these cases in the perfect mid-sagittal plane the nasal bone may erroneously be considered to be absent. PMID- 15816009 TI - Analysis of residues of imidacloprid in tobacco by high-performance liquid chromatography with liquid-liquid partition cleanup. AB - A practical method for analysis of residues of imidacloprid in baked tobacco leaves has been developed using HPLC with liquid-liquid partition clean-up. Imidacloprid was extracted with ethyl acetate under ultrasound, and cleaned up by liquid-liquid partition with 50 g litre(-1) aqueous sodium chloride followed by dichloromethane. The water-soluble and fat-soluble components in tobacco, which interfere with imidacloprid in HPLC, were removed. The separation was performed on a Supelco LC-18 column (250 mm x 4.6 mm ID) with a mobile phase of acetonitrile + 5 mM ammonium acetate (20 + 80 by volume) at a flow-rate of 1 ml min(-1). The baseline separation between imidacloprid and the tobacco blank was achieved within 10 min. With a detection wavelength at 270 nm, the limit of quantitation was 0.04 mg kg(-1). The recovery ranged from 89.8 to 95.4% and the RSDs were less than 2.3%. The proposed method was successfully employed for the determination of imidacloprid residues in 300 samples of flue-cured tobacco leaves. PMID- 15816010 TI - Anatoxins and degradation products, determined using hybrid quadrupole time-of flight and quadrupole ion-trap mass spectrometry: forensic investigations of cyanobacterial neurotoxin poisoning. AB - The potent neurotoxins from cyanobacteria, anatoxin-a (AN), its methyl analogue, homoanatoxin-a (HMAN), and their degradation products, have been studied using nano-electrospray hybrid quadrupole time-of-flight mass spectrometry (QqTOF-MS). The anatoxin degradation products, which are readily produced in vivo by either reduction or epoxidation, were also examined in this study. The high mass accuracy QqTOF-MS data was used to confirm formula assignments for major product ions and quadrupole ion-trap (QIT)-MS was used to construct fragmentation pathways for anatoxins. Significant differences between these fragmentation pathways were observed. Comparisons between the spectra of compounds that differ in side-chain length (the AN and HMAN series) were used to identify ions that are characteristic of the homologues. The application to forensic samples in which the principal neurotoxin had undergone rapid biodegradation has been demonstrated and used to confirm anatoxin poisoning of dogs. PMID- 15816011 TI - Pattern of cannabis use in ecstasy polydrug users: moderate cannabis use may compensate for self-rated aggression and somatic symptoms. AB - Cannabis is one of the most common 'co-drugs' for ecstasy users. The aim of the present study was to explore self-reported psychobiological problems in ecstasy polydrug users in relation to their pattern of cannabis use. Two hundred and eighty ecstasy polydrug users were allocated into five cannabis groups according to the frequency of their cannabis use. The control group comprised 121 alcohol tobacco users. There were no significant group differences with regard to age, diagnosed family psychiatric history and level of self-rated stress experienced during 6 months prior to the study. The present study produced three main findings: (a) Ecstasy users with no concomitant use of cannabis displayed more self-rated aggression and somatic symptoms compared with ecstasy users who were smoking cannabis on a monthly or weekly basis. (b) Ecstasy users who reported heavy cannabis use in the past displayed higher paranoid symptoms compared with ecstasy weekly and daily cannabis users. (c) Former heavy cannabis users were the most likely to complain of a variety of ecstasy related long-term problems. In conclusion, moderate cannabis use may help to ameliorate or mask MDMA-induced aggressivity and somatic symptoms. However, this study confirms that heavy cannabis and ecstasy use is associated with several psychobiological problems, which may emerge after a period of abstinence from both drugs. PMID- 15816012 TI - Methods for capture-recapture analysis when cases lack personal identifiers. AB - Methods for estimating the size of a closed population from a capture-recapture study require the availability of unique identifiers on each of two lists. These identifiers are used to identify the number of individuals appearing on both lists. When the number of individuals appearing on both lists cannot be determined with certainty from the data, matching between the lists is problematic. In this paper, we develop a weighted estimator to account for all possible matches between two lists. A bootstrap procedure is proposed for estimation. To illustrate the methods, we used two lists that recorded New York State (NYS) hospitalizations due to pertussis in 1996 to estimate the number of persons hospitalized for pertussis in NYS that year. PMID- 15816013 TI - Prostaglandin D2 affects the differentiation and functions of human dendritic cells: impact on the T cell response. AB - The local environment in which dendritic cells (DC) differentiate is important for the acquisition of their immunostimulatory properties. Since prostaglandin D(2) (PGD(2)), a major prostanoid produced during inflammatory reactions, is involved in the control of immune responses, its effect on the differentiation and functions of human monocyte-derived dendritic cells (MDDC) was studied. We show that DC differentiated in the presence of PGD(2) (PG/DC) have an unusual phenotype, with modifications in the expression of molecules involved in antigen (Ag) capture and presentation, leading to higher endocytic and Ag-processing activities. However, under conditions that necessitated Ag processing and presentation, PG/DC have an impaired ability to stimulate naive T cells, whereas superAg-pulsed DC efficiently promote their proliferation. Upon lipopolysaccharide or TNF-alpha/IL-1beta stimulation, PG/DC phenotypically mature but produce abnormal amounts of immunoregulatory cytokines (decreased IL-12p70/IL 10 ratio). Moreover, mature PG/DC fail to up-regulate the chemokine receptor CCR7 and show an impaired migration towards its ligand CCL19. Finally, PG/DC favor the differentiation of naive T cells toward Th2 cells, an effect dependent on IL-10 and inducible costimulator ligand expression by DC. Most of the herein described effects of PGD(2) on MDDC can be reproduced, usually with a higher efficacy, with a selective D prostanoid receptor (DP)1, but not DP2, agonist. Taken as a whole, these results demonstrate that PGD(2) impacts DC differentiation and functions, and extend the concept that it exerts important roles in immunity. PMID- 15816014 TI - CD8 T cell priming by B lymphocytes is CD4 help dependent. AB - While it is generally accepted that B lymphocytes can present antigen and activate CD4 T cells, priming of CD8 T cells by B lymphocytes remains controversial. Recently, we showed that mice injected with genetically programmed B lymphocytes generate antigen specific CD4 and CD8 T cell responses in vivo that could also be induced in mice lacking functional dendritic cells. To gain further insights into the requirements for T cell priming by antigen-presenting B lymphocytes, in vitro experiments were performed using ovalbumin (OVA) and OVA specific TCR-transgenic CD4 and CD8 T cells. We found that while B lymphocytes can directly prime CD4 T cells, the activation of CD8 T cells requires T cell help. Transfer experiments show that help can either be contact dependent or be mediated by soluble factors in the supernatants of activated OVA-specific CD4 T cells. Furthermore, the effect of activated CD4 T cells can be replaced by soluble recombinant IL-4. Collectively, the data show the existence of different requirements for priming of CD4 and CD8 T cells and point to the previously unappreciated fact that the induction of CD8 T cell responses by B lymphocytes requires T cell help. PMID- 15816015 TI - The human beta-glucan receptor is widely expressed and functionally equivalent to murine Dectin-1 on primary cells. AB - We identified the C-type-lectin-like receptor, Dectin-1, as the major receptor for fungal beta-glucans on murine macrophages and have demonstrated that it plays a significant role in the cellular response to these carbohydrates. Using two novel, isoform-specific mAb, we show here that human Dectin-1, the beta-glucan receptor (betaGR), is widely expressed and present on all monocyte populations as well as macrophages, DC, neutrophils and eosinophils. This receptor is also expressed on B cells and a subpopulation of T cells, demonstrating that human Dectin-1 is not myeloid restricted. Both major functional betaGR isoforms - betaGR-A and betaGR-B - were expressed by these cell populations in peripheral blood; however, only betaGR-B was significantly expressed on mature monocyte derived macrophages and immature DC, suggesting cell-specific control of isoform expression. Inflammatory cells, recruited in vivo using a new skin-window technique, demonstrated that Dectin-1 expression was not significantly modulated on macrophages during inflammation, but is decreased on recruited granulocytes. Despite previous reports detailing the involvement of other beta-glucan receptors on mature human macrophages, we have demonstrated that Dectin-1 acted as the major beta-glucan receptor on these cells and contributed to the inflammatory response to these carbohydrates. PMID- 15816016 TI - IL-4Ralpha signaling is important for CD8+ T cell cytotoxicity in the absence of CD4+ T cell help. AB - Cytotoxic CD8(+) T cells are key mediators of viral clearance during primary infection through their production of IFN-gamma and lysis of virally infected cells. Comparatively, the cytokines IL-4 and IL-13 are typically associated with the development of Th2 immune responses against allergens and parasites, while their influence on cytotoxic CD8(+) T cell responses is controversial. We have investigated the roles of IL-4 and IL-13 in the development of CD8(+) T cell responses against influenza infection. We show that in the absence of either IL-4 or IL-13, CD8(+) T cells proliferated and a normal secondary cytotoxic response developed in vitro. In striking contrast, the absence of IL-4Ralpha resulted in impaired ex vivo proliferation and consequently no secondary CTL activity, whereas the in vivo response appeared normal. We show that the presence of CD4(+) T cell help, or the addition of exogenous IL-2 in vitro, restored the response. Taken together, this work reveals previously unrecognized in vivo redundancies between IL-4, IL-13 and IL-2 during immune responses against influenza virus. PMID- 15816017 TI - Characterization and PCR-based detection of benzimidazole-resistant isolates of Monilinia laxa in California. AB - Monilinia laxa is a pathogen of brown rot of stone fruit and almond in California, causing blossom blights and fruit rots. In this study, low-level resistance to the benzimidazole fungicides benomyl and thiophanate-methyl was detected in field isolates of M laxa collected from stone fruits and almonds in California. Low-resistant (LR) isolates grew in potato dextrose agar (PDA) plates amended with benomyl and thiophanate-methyl at 1 and 5 microg ml(-1), respectively, but not in plates amended with benomyl at 5 microg ml(-1) or thiophanate-methyl at 50 microg ml(-1). The benzimidazole LR isolates were characterized by temperature sensitivity and the DNA sequence of the beta-tubulin gene. The LR isolates showed high-temperature sensitivity, being sensitive to 1 microg ml(-1) of benomyl at 28 degrees C but resistant at 8-24 degrees C. Analysis of the DNA sequence of the beta-tubulin gene showed that the LR isolates had a point mutation at the amino-acid position 240, causing substitution of leucine by phenylalanine. Based on the point mutation, a pair of allele-specific PCR primers was developed for rapid detection of LR isolates of M laxa. In addition, a pair of PCR primers specific to M laxa was developed on the basis of the differences in the DNA sequence of the intron 6 of beta-tubulin gene from M laxa, M fructicola and other fungal species. The primer pair amplified the expected 376-bp DNA fragment from all M laxa isolates tested, but not from 14 other fungal species isolated from stone fruit and almond crops. The restriction endonuclease BsmA I recognized the sequence GTCTCC in the PCR products from sensitive (S) isolates only, but not the GTTTCC sequence in the PCR products from LR isolates. The endonuclease digested the 376-bp PCR products from S isolates to produce two bands (111 and 265 bp) on agarose gels. Thus, both allele-specific PCR and the PCR-restriction fragment length polymorphism (PCR-RFLP) methods could be useful for rapidly detecting benzimidazole-resistant isolates of M laxa from stone fruit and almond crops in California. PMID- 15816018 TI - A study of the disappearance of pesticides during composting using a gas chromatography-tandem mass spectrometry technique. AB - A simple method for multi-residue analysis of pesticides by low-pressure gas chromatography-tandem mass spectrometry (LP-GC-MS-MS) has been validated in compost samples. The pesticide residues were extracted from the lyophilised samples with organic solvent by stirring. No sample clean-up was required prior to the analysis. The method was applied to determine the fate of two organochlorine pesticides (lindane and endosulfan) and two organophosphates (malathion and chlorpyrifos-methyl) during the composting process in the reactor of a pilot plant. Malathion, chlorpyrifos-methyl and lindane residues almost fully disappeared after 8 days of maturation in the reactor, while endosulfan residues were only partially degraded. Alongside this, a study of natural disappearance of the pesticides was carried out in the laboratory and a characteristic time profile was traced for each pesticide class. PMID- 15816019 TI - Incompatibility of metam sodium with halogenated fumigants. AB - Metam sodium (metam) is a widely used soil fumigant. Combined application of metam and other available fumigants is intended to produce synergic pesticidal effects for a broad spectrum of pest control in soil fumigation. This study aimed to test the compatibility of metam with the halogenated fumigants 1,3 dichloropropene (1,3-D), chloropicrin, methyl bromide, methyl iodide and propargyl bromide. Halogenated fumigants and metam were spiked simultaneously into organic solvents, water and moist soils, and metam-induced degradation of these halogenated chemicals was evaluated. In all three media, the halogenated fumigants were incompatible with metam and degraded via rapid chemical reactions. The degradation rate varied with halogenated fumigant species and increased as the amount of metam present was increased. In moist soil, 15-95% of the halogenated fumigants were decomposed within 72 h by metam at a 1:1 molar ratio. Combined application of Telone C-35 (62.5% 1,3-D + 35% chloropicrin) at 265 mg kg(-1) and Vapam (42% metam) at 567 mg kg(-1) in soil resulted in complete disappearance of the applied chloropicrin and 20-38% of the 1,3-D within 8 h. The results suggest that simultaneous application of halogenated fumigants and metam at the same soil depth will not maximize pest control. In practice, sequential treatment of soil or application at different soil depths is recommended when these two types of fumigants are used in combination. PMID- 15816020 TI - Is fetal cerebral vascular resistance affected by the presence of nuchal cord(s) in the third trimester of pregnancy? AB - OBJECTIVE: To assess whether fetal cerebral vascular resistance is affected by the presence of nuchal cord(s) in the third trimester. METHODS: A set of 115 patients with well-established dates and singleton, appropriate for gestational age (GA), non-anomalous fetuses with nuchal cord(s) diagnosed with prenatal color Doppler imaging, between 28 and 41 weeks and 115 controls matched for GA, were studied. Patients with hypertension, diabetes and autoimmune conditions were excluded. Doppler indices were obtained from the umbilical artery (UA) and the fetal middle cerebral artery (MCA) upon initial diagnosis of the nuchal cord. RESULTS: Of 115 GA-matched pairs of pregnancies, 103 fetuses had a single nuchal cord and 12 a double nuchal cord. No significant differences were noted in patient age, gravidity, parity, sonographically estimated fetal weight (SEFW), and growth centile at ultrasonographic diagnosis. Mean UA systolic/diastolic ratio (S/D) was 2.28 +/- 0.32 and 2.36 +/- 0.33 (P = 0.03) and UA resistance index (RI) 0.55 +/- 0.06 and 0.57 +/- 0.06 (P = 0.02) among study and controls, respectively. Mean fetal MCA S/D and RI did not differ significantly between the two groups (fetal MCA S/D 5.01 +/- 1.28 and 5.04 +/- 1.71 (P = 0.86), and mean fetal MCA RI 0.78 +/- 0.06 and 0.78 +/- 0.05 (P = 0.88)). No significant differences were noted in GA at delivery, incidence of meconium-stained amniotic fluid, birth weight, mode of delivery, neonatal gender, 1 and 5 minute Apgar scores, or UA pH and base excess, between study and control patients. CONCLUSION: Fetal cerebral vascular resistance is not affected by the presence of nuchal cord(s) in the third trimester of pregnancy. PMID- 15816021 TI - Correlation between ultrasound and anatomical findings in fetuses with lower urinary tract obstruction in the first half of pregnancy. AB - OBJECTIVE: The prognosis of fetal lower urinary tract obstruction (LUTO) depends upon renal function and also upon the underlying etiology. Precise identification of the latter remains a challenge antenatally. Our objective was to examine the underlying pathology in male fetuses with sonographic evidence of severe and isolated LUTO. METHODS: Detailed postmortem examination was carried out after termination of pregnancy in 24 male fetuses presenting before 25 weeks of gestation with ultrasound evidence of isolated severe LUTO. RESULTS: All fetuses had megacystis and hyperechogenic kidneys. There was anhydramnios/oligohydramnios and pelvicalyceal dilatation in 20 and 15 cases, respectively. Posterior urethral valves (PUV) were suspected antenatally in 20 cases and urethral atresia was not suspected antenatally. However, postmortem examination of the urethra demonstrated atresia in six cases, severe stenosis in eight cases, PUV in nine cases and an apparently normal urethra in one case. Renal dysplasia was found in all cases but one. Urethral atresia was the most common urethral anomaly at 12-17 weeks. Hydronephrosis was more frequent in cases with PUV (8/9) and urethral stenosis (6/8) than with urethral atresia (0/6). In LUTO presenting in the first and second trimester, hyperechogenic kidneys were predictive of renal dysplasia in 95% of cases. The association of a sagittal diameter of the bladder of at least 40 mm with hydronephrosis before 28 weeks was predictive of PUV with a positive (PPV) and negative (NPV) predictive value of 44.4% and 66.6%, respectively. Absence of hydronephrosis and a sagittal diameter of the bladder of less than 40 mm were predictive of urethral atresia or stenosis with a PPV and NPV of 100% and 47.6%, respectively. The absence of hydronephrosis was predictive of urethral atresia with a PPV and NPV of 66.6% and 100%, respectively. CONCLUSION: LUTO in a male fetus presenting with megacystis in the first or second trimester of pregnancy is as likely to reflect urethral atresia or stenosis as it is PUV. The size of the bladder and the presence of hydronephrosis should be considered in order to improve prenatal diagnosis and counseling. PMID- 15816022 TI - Enzyme immobilization on ultrafine cellulose fibers via poly(acrylic acid) electrolyte grafts. AB - Ultrafine cellulose fiber (diameter 200-400 nm) surfaces were grafted with polyacrylic acid (PAA) via either ceric ion initiated polymerization or methacrylation of cellulose with methacrylate chloride (MACl) and subsequent free radical polymerization of acrylic acid. PAA grafts by ceric ion initiated polymerization increased with increasing reaction time (2-24 h), monomer (0.3-2.4 M), and initiator (1-10 mM) concentrations, and spanned a broad range from 5.5 850%. PAA grafts on the methacrylated cellulose fibers also increased with increasing molar ratios of MACl to cellulosic hydroxyl groups (MACl/OH, 2-6.4) and monomer acrylic acid (AA) to initiator potassium persulfate (KPS) ratios ([AA]/[KPS], 1.5-6), and were in a much narrower range between 12.8% and 29.4%. The adsorption of lipase (at 1 mg/ml lipase and pH 7) and the activity of adsorbed lipase (pH 8.5, 30 degrees C), in both cases decreased with increasing PAA grafts. The highest adsorption and activity of the lipase on the ceric ion initiated grafted fibers were 1.28 g/g PAA and 4.3 U/mg lipase, respectively, at the lowest grafting level of 5.5% PAA, whereas they were 0.33 g/g PAA and 7.1 U/mg lipase, respectively, at 12.8% PAA grafts on the methacrylated and grafted fibers. The properties of the grafted fibers and the absorption behavior and activity of lipase suggest that the PAA grafts are gel-like by ceric-initiated reaction and brush-like by methacrylation and polymerization. The adsorbed lipase on the ceric ion-initiated grafted surface possessed greatly improved organic solvent stability over the crude lipase. The adsorbed lipases exhibited 0.5 and 0.3 of the initial activity in the second and third assay cycles, respectively. PMID- 15816023 TI - Online monitoring and characterization of flocculating yeast cell flocs during continuous ethanol fermentation. AB - Both intrinsic and observed kinetic investigations for those ethanol fermentations using self-flocculated yeast strains have been hindered by the lack of real online monitoring techniques and proper characterization methods for the flocs. An optical detecting technique, the focused beam reflectance measurement probe developed by Lasentec (Redmond, WA) was inserted into a fermentor to monitor the floc chord length distributions. Using a simulating system composed of the floc-buffer suspensions, the total floc chord length counts per second were directly correlated with the floc biomass concentrations so that the floc biomass concentrations can be in situ detected. Furthermore, a characterization method of the flocs was established by properly weighted treatments of the detected floc chord length distributions. When a real yeast floc ethanol fermentation system was detected during its intrinsic kinetic investigations in which the floc size needed to be controlled at a level of micrometer scale to eliminate inner mass transfer limitations, it was found and validated that CO(2) produced during fermentation exerted significant disturbances. By applying 1/length-weighted treatment, these disturbances were effectively overcome. PMID- 15816024 TI - A bio-recognition device developed onto nano-crystals of carbonate apatite for cell-targeted gene delivery. AB - The DNA delivery to mammalian cells is an essential tool for analyzing gene structure, regulation, and function. The approach holds great promise for the further development of gene therapy techniques and DNA vaccination strategies to treat and control diseases. Here, we report on the establishment of a cell specific gene delivery and expression system by physical adsorption of a cell recognition molecule on the nano-crystal surface of carbonate apatite. As a model, DNA/nano-particles were successfully coated with asialofetuin to facilitate uptake by hepatocyte-derived cell lines through the asialoglycoprotein receptor (ASGPr) and albumin to prevent non-specific interactions of the particles with cell-surface. The resulting composite particles with dual surface properties could accelerate DNA uptake and enhance expression to a notable extent. Nano-particles coated with transferrin in the same manner dramatically enhanced transgene expression in the corresponding receptor-bearing cells and thus our newly developed strategy represents a universal phenomenon for anchoring a bio-recognition macromolecule on the apatite crystal surface for targeted gene delivery, having immediate applications in basic research laboratories and great promise for gene therapy. PMID- 15816026 TI - Selective separation of beta-lactoglobulin from sweet whey using CGAs generated from the cationic surfactant CTAB. AB - The selective separation of whey proteins was studied using colloidal gas aphrons generated from the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). From the titration curves obtained by zeta potential measurements of individual whey proteins, it was expected to selectively adsorb the major whey proteins, i.e., bovine serum albumin, alpha-lactalbumin, and beta-lactoglobulin to the aphrons and elute the remaining proteins (lactoferrin and lactoperoxidase) in the liquid phase. A number of process parameters including pH, ionic strength, and mass ratio of surfactant to protein (M(CTAB)/M(TP)) were varied in order to evaluate their effect on protein separation. Under optimum conditions (2 mmol/l CTAB, M(CTAB)/M(TP) = 0.26-0.35, pH 8, and ionic strength = 0.018 mol/l), 80-90% beta-lactoglobulin was removed from the liquid phase as a precipitate, while about 75% lactoferrin and lactoperoxidase, 80% bovine serum albumin, 95% immunoglobulin, and 65% alpha-lactalbumin were recovered in the liquid fraction. Mechanistic studies using zeta potential measurements and fluorescence spectroscopy proved that electrostatic interactions modulate only partially the selectivity of protein separation, as proteins with similar surface charges do not separate to the same extent between the two phases. The selectivity of recovery of beta-lactoglobulin probably occurs in two steps: the first being the selective interaction of the protein with opposite-charged surfactant molecules by means of electrostatic interactions, which leads to denaturation of the protein and subsequent formation and precipitation of the CTAB-beta-lactoglobulin complex. This is followed by the separation of CTAB-beta-lactoglobulin aggregates from the bulk liquid by flotation in the aphron phase. In this way, CGAs act as carriers which facilitate the removal of protein precipitate. PMID- 15816028 TI - Application of fluorescence recovery after photobleaching to diffusion of a polyelectrolyte in a multilayer film. AB - The diffusion coefficient, D, and the proportion of mobile molecules, p, is measured for a fluorescently labeled polyelectrolyte in a multilayer film using fluorescence recovery after photobleaching (FRAP). The film was composed of poly(L-lysine) (PLL) and hyaluronan (HA). The labeled polyelectrolyte (PLL(FITC)) was either deposited on top of the film or embedded within it. A circular area of diameter approximately 60 microm was bleached using a confocal laser scanning microscope. Because molecules do already diffuse during the bleaching step, the initial light intensity profile is not characteristic of a uniformly bleached circular area. A formalism is developed in which a simple mathematical representation of a measured profile serves as starting profile. This radial distribution is introduced in the equation describing the time evolution of the labeled molecule concentration under the hypothesis that the recovery results from pure, two-dimensional Brownian diffusion of the mobile molecules according to Fick's law. The analysis of a series of images taken at successive times after bleaching (up to approximately one hour) leads to estimates of D of the order of 0.2 microm(2) s(-1) for labeled molecules deposited on top of the film and a 5 fold smaller value for the molecules embedded in the film. However, p is remarkably insensitive to the position in the multilayer architecture (p approximately 0.40). PMID- 15816029 TI - Hyperplastic perilobar nephroblastomatosis: long-term survival of 52 patients. AB - BACKGROUND: This study provides insight into the clinical behavior, diagnostic complexities, and long-term management of patients with hyperplastic perilobar nephroblastomatosis (HPLN). PROCEDURE: Fifty-two patients with HPLN with available long-term follow-up were retrospectively analyzed for pathologic, radiologic, and clinical features. RESULTS: The mean age at diagnosis was 16 months; the lesions were bilateral in 49 patients. Of 33 patients who initially underwent diagnostic biopsy and adjuvant chemotherapy, 18 (55%) developed Wilms tumor (WT) at a mean of 35 months from diagnosis. Of 16 patients whose initial therapy included nephrectomy and adjuvant therapy, three (19%) developed WT at a mean of 36 months from diagnosis. All three patients who underwent initial diagnostic biopsy and received no adjuvant therapy during their initial course developed WT 4, 4, and 10 months following diagnosis. 24/52 patients developed either a single (13 patients) or multiple (11 patients) WT throughout their course; 8/24 (33%) of WT were anaplastic. The time from initial diagnosis to the development of the last WT ranged from 13 to 116 months (mean 42 months). Three children with HPLN died of WT at 3, 5, and 6 years of age; 2/3 were anaplastic. CONCLUSIONS: HPLN is a self-limited, pre-neoplastic proliferative process associated with a high risk of developing WT. The accurate diagnosis and the choices of therapy during the often-complex course of HPLN depend on the availability and accurate interpretation of a combination of pathologic, radiologic, and clinical information. When such information is appropriately obtained, the long-term survival of patients with HPLN is excellent. PMID- 15816030 TI - Embedding of large specimens in glycol methacrylate: prerequisites for multi signal detection and high-resolution imaging. AB - Acrylic resin mixtures are commonly used to study microscopic sections of biological specimens, giving the advantage of good morphological preservation. Existing embedding protocols, however, are suitable for tissue blocks, not exceeding 1 mm in thickness. We have developed a protocol to embed larger specimens (up to 2 cm(3)) in Technovit 8100. This medium allowed us to perform classic histological (trichrome), silver, as well as immunohistochemical staining, needed for multi-signal detection at high-resolution imaging to reconstruct a three-dimensional interpretation of a serially sectioned muscle. The technique was applied to reconstruct the semitendinosus muscle of a fetal pig, 44 days post conception, featuring connective tissue, intramuscular nerves, blood vessels, and muscle fibre types. For the reconstruction, a technique was used that enabled us to insert high-resolution images of histological details into low-resolution images of the entire muscle. PMID- 15816031 TI - Effect of Eupatorium adenophorum Spreng leaf extracts on the mustard aphid, Lipaphis erysimi Kalt: a scanning electron microscope study. AB - Scanning electron microscopic study revealed some remarkable adverse effects of Eupatorium adenophorum Spreng leaf extract on cuticular and antennal sensilla of the mustard aphid, Lipaphis erysimi Kalt. The study appears to be the first of its kind in providing evidence on toxic property of the plant on certain agricultural pests, although the indigenous tribes of the region know the insecticidal property of the plant for a long time. The study further provides indications regarding the neurotoxic nature of the leaf extract since it has caused abnormalities in the sensory structures. The presence of prominent swellings in some sensilla and shrinkage in others suggests that the leaf extract affects different systems through different mechanisms. The possible use of the plant as a potential biopesticide against certain insect pests is discussed. PMID- 15816033 TI - Interfaces between dendritic cells, other immune cells, and nerve fibres in mouse Peyer's patches: potential sites for neuroinvasion in prion diseases. AB - In this study, we examined where immune cells and nerve fibres are located in mouse Peyer's patches, with a view to identifying potential sites for neuroinvasion by prions. Special attention was paid to dendritic cells, viewed as candidate transporters of infectious prion. Double immunofluorescence labellings with anti-CD11c antibody and marker for other immune cells (B cells, T cells, follicular dendritic cells) were carried out and analysed by confocal microscopy on Peyer's patch cryosections. To reveal the extensive ganglionated networks of the myenteric and submucosal plexi and the sparse meshworks of nerve strands, we used antibodies directed against different neurofilament subunits or against glial fibrillary acidic protein. In the suprafollicular dome, dendritic cells connect, via their cytoplasmic extensions, enterocytes with M cells of the follicle-associated epithelium. They are also close to B and T cells. Nerve fibres are detected in the suprafollicular dome, notably in contact with dendritic cells. Similar connections between dendritic cells, T cells, and nerve fibres are seen in the interfollicular region. Germinal centres are not innervated; inside them dendritic cells establish contacts with follicular dendritic cells and with B cells. After immunolabelling of normal prion protein, dendritic cells of the suprafollicular dome are intensely positive labelled. PMID- 15816034 TI - Distribution of NGF and NT-3-like protein immunoreactivity in the teleost kidney. AB - By means of immunochemistry and immunohistochemistry, we investigated in the kidney of freshwater and marine teleostean species for the presence and localization of three neurotrophins: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin (NT)-3. In both species studied, NGF like and NT-3-like immunoreactivity were present in the kidney with different distribution patterns, while BDNF-like immunoreactivity was never detected. In goldfish, NGF-like and NT-3-like immunoreactivity were identified extensively in cells along part of the arterial branches adjacent to the afferent arterioles. In scorpion fish, NGF-like and NT-3-like immunoreactive cells were observed both on afferent arterioles and on adjacent secondary branches derived from renal arteries. No immunoreactivity was detected in other renal structures. A staining pattern of immunoreactivity similar to that obtained for NGF and NT-3 was detected utilizing S100 antibody as a juxtaglomerular (JG) cell marker. Double immunolabellings NGF/S100 and NT-3/S100 evidenced the coexistence of neurotrophin like proteins and S100-like protein in the same immunoreactive cells, thus identifying them as juxtaglomerular cells. Western blot analysis revealed the presence of molecules immunoreactive to NGF and NT-3, whose molecular weights were very similar to those of the corresponding mammalian neurotrophins. These findings extend the presence and distribution of NGF-like and NT-3-like IR in the kidney to teleost species, suggesting a probable participation of these proteins in the renal functions of freshwater and marine teleosts. PMID- 15816035 TI - Inflammatory cell recruitment and adhesion to methyl-terminated self-assembled monolayers: effect of implantation time. AB - The contribution of methyl groups in implant-triggered inflammation was investigated in vivo using self-assembled monolayers (SAMs) of alkanethiols on gold. The CH(3)-coated implants were inserted in an air-pouch cavity induced in BALB/c mice. The in situ inflammatory response was monitored 24, 48, and 72 hours later. Inflammatory cells recovered from the air pouches were counted and observed by light microscopy. The cellularity of the implant surfaces was defined by scanning electron microscopy (SEM). In comparison with gold implants, the CH(3)-coated SAMs recruited a significantly higher number of inflammatory cells. Polymorphonuclear leukocytes (PMN) were more numerous than mononuclear cells (Mo) in the exudates recovered from the air pouches with CH(3)-coated SAMs. The opposite PMN/Mo proportion was observed in air pouches of the two control groups (mice receiving gold implants or sham-operated animals). A low density of adherent cells was seen on CH(3)-coated implants, with no significant quantitative differences during the time course of the study. In contrast, the gold-coated surfaces were covered with numerous cells during all of the 3 days of the inflammation. In conclusion, implants with CH(3) surfaces are likely to induce PMN-dominated local acute inflammation but these surfaces are not associated with a significant adherence of leukocytes to the implant. PMID- 15816036 TI - Spanish colonial effects on Native American mating structure and genetic variability in northern and central Florida: Evidence from Apalachee and western Timucua. AB - Standard population genetic analyses are implemented for a series of precontact and contact period samples from central and northern Florida to investigate changes in genetic variability and population affinity coincident with the establishment of Spanish missions during the 17th century. Estimates of F(ST) based on odontometric data indicate limited heterogeneity for the Apalachee samples, suggestive of some degree of within-group endogamy for this ethnic group prior to contact. This corresponds well with ethnohistoric reconstructions indicating that Apalachee were populous, partially linguistically isolated from its neighbors, and involved in persistent cycles of warfare with neighboring groups. Estimates of extralocal gene flow for the Apalachee samples indicate limited initial changes in the mating structure of these populations. After 1650, however, extralocal gene flow increases, consistent with evidence for dramatic population movements throughout northern Florida and increased Spanish presence in the province, particularly at the mission of San Luis. Inclusion of non Apalachee outgroups does not increase estimates of genetic heterogeneity, as was expected based on ethnohistoric data. The pattern of genetic distances suggests a biological division between north and south Florida population groups, consistent with archaeological and ethnohistoric data, and similarly indicates some distinction between precontact and postcontact local groups. Differential extralocal gene flow experienced by pre-1650 Apalachee and Timucua populations suggests localized mission experience. The Apalachee, with large, dense populations, experienced limited initial changes in genetic diversity or mating structure. However, after 1650 they were apparently involved in a much more expansive mating network that may have included Spaniards and immigrant Native American groups to the region. These results are in contrast to the mission experience of the Guale Indians of the Georgia coast. PMID- 15816037 TI - FIB cross-sectioning of a single rapidly solidified hypereutectic Al-Si powder particle for HRTEM. AB - A creative technique of in-situ focused ion beam (FIB) extraction was introduced to prepare a gas atomized rapidly solidified hypereutectic Al-Si single particle's cross-section for High Resolution Transmission Electron Microscopy (HRTEM) analysis. This preparation technique may be employed to characterize very inimitable samples that are abnormally wrought or intricate to prepare through traditional techniques. TEM results revealed that a gas-atomization/rapid solidification process leads to a homogeneous dispersion of 50-100-nm Si phase in the Al matrix. Stacking faults and dislocations are observed in the microstructure and will ultimately lead to the increased strength in a resultant bulk material manufactured from this powder to be further examined. PMID- 15816038 TI - Pigment epithelium-derived factor induces the production of chemokines by rat microglia. AB - Many studies have shown that pigment epithelium-derived factor (PEDF) has neurotrophic effects on retinal cells and hippocampal, spinal cord, and cerebellar granule cell neurons, but much less work has examined the effects of PEDF on glia. In this study, we show that PEDF changes microglial morphology within 1 h of exposure, to a more deactivated form, while having no effect on the expression of such activation markers as OX-42 and ED-1. In contrast, urea activates acid phosphatase, and PEDF blocks that activation. PEDF also activates NFkappaB, accompanied by the induction of mRNAs and proteins for the chemokines macrophage inflammatory protein-1alpha (MIP-1alpha, MIP-2, and MIP-3alpha. All the chemokines stimulate acid phosphatase activity, and high doses of MIP-2 and MIP-3alpha), alter the morphology of the microglia at 1 h after treatment. These results suggest that the use of PEDF for clinical treatments, such as for retinal neovascularization, brain injury, or ischemia, should be undertaken with caution because of the possibility of induction of inflammation caused by microglial or other immune cell migration in response to the chemokines induced by PEDF. PMID- 15816039 TI - Unique ramus anatomy for Neandertals? AB - The ramus of Neandertal mandibles is said to show a suite of uniquely Neandertal character states that demonstrate the independent course of Neandertal evolution. This is the latest of numerous attempts to define cranial and mandibular autapomorphies for Neandertals. We examine variation in the four presumably autapomorphic ramal features and show they are neither monomorhic within Neandertals (to the contrary Neandertals are at least as variable as other human samples) nor unique to Neandertals, since they regularly appear in populations predating and postdating them. Neandertals differ from other human populations, both contemporary and recent, but the question of whether this fact reflects a divergent evolutionary trajectory must be addressed by the pattern of differences. In this case, as in the other attempts to establish Neandertal autapomorphies, rather than showing restricted variation and increased specialization, the Neandertal sample shows that the range of human variation in the recent past encompasses, and in some cases exceeds, human variation today, even in the very features claimed to be autapomorphic. PMID- 15816047 TI - Structure-activity relationships of inverse agonists for G-protein-coupled receptors. AB - It has been recently established that G-protein-coupled receptors (GPCRs) can be constitutively active, i.e., they can be active in the absence of an agonist. This activity can be inhibited by so-called inverse agonists. For a number of GPCRs, such inverse agonists have been developed and studied, now enabling for the first time a study into their structure-activity relationships. PMID- 15816048 TI - The impact of a delay in initiating radiation therapy on prostate-specific antigen outcome for patients with clinically localized prostate carcinoma. AB - BACKGROUND: To determine whether a delay in initiating external beam radiation therapy (RT) following diagnosis could impact prostate-specific antigen (PSA) outcome for patients with localized prostate cancer, 460 patients, who received 3D conformal RT to a median dose of 70.4 Gy for clinically localized prostate cancer between 1992 and 2001, were studied. METHODS: The primary endpoint was PSA failure (American Society for Therapeutic Radiology and Oncology definition). Estimates of PSA control were made using the Kaplan-Meier method. Delay was defined as the time between diagnosis and the start of RT. Risk groups were defined based on known predictors of PSA outcome, namely, baseline PSA level, clinical T-category, Gleason score, and percentage of biopsy cores positive for tumor. Cox multivariate regression analysis was used to determine the ability of treatment delay to predict time to PSA failure after adjusting for the other known predictors. RESULTS: Treatment delay independently predicted time to PSA failure following diagnosis for high-risk (Adjusted Hazard Ratio = 1.08 per month; P = 0.029) but not low-risk patients (P = 0.31). Patients with high-risk disease (n = 240) had 5-year estimates of PSA failure-free survival of 55% versus 39% (Plog-rank = 0.014) for those with delay < 2.5 months versus > or = 2.5 months respectively. The median delay was 2.5 months. CONCLUSIONS: Treatment delay adversely affected PSA outcome for high-risk patients but not for low-risk patients following RT. PMID- 15816049 TI - Postoperative complications of salvage total laryngectomy. AB - BACKGROUND: The objectives of the current study were to report the incidence of postoperative complications for salvage total laryngectomy (STL) compared with primary total laryngectomy (PTL) and to identify patient and tumor-related factors predictive of postoperative complications. METHODS: A sample of 183 patients who had received a total laryngectomy were identified from an existing database of 662 patients treated for squamous cell carcinoma of the larynx. PTL and STL were performed in 113 and 70 patients, respectively. Initial therapy in the patients who required salvage surgery included radiotherapy (RT) in 32 (46%) and chemoradiotherapy (CTRT) in 38 (54%). Postoperative complications were recorded for each group and categorized into local, swallowing, airway, and systemic complications. Postoperative complication rates for STL after RT and CTRT were compared with those after PTL by univariate analysis. Patient and tumor related predictors of complications were identified by univariate and multivariate analyses. RESULTS: The overall mortality rate was 0.5%. Forty percent of all patients developed a postoperative complication after total laryngectomy. Local complications, which were the most frequent, occurred in 52 (28%) patients. Pharyngocutaneous fistula occurred in 31 (17%) patients. Statistical analysis showed that there was a greater number of patients with local wound (45% vs. 25%, P = 0.02) and fistula complications (32% vs. 12%, P = 0.012) in the STL-CTRT group compared with the primary laryngectomy group. Multivariate analysis showed that primary CTRT was an independent predictor of local complications and pharyngocutaneous fistula. CONCLUSIONS: Salvage laryngectomy was more frequently associated with postoperative complications after CTRT compared with PTL. Problems related to local wound healing, especially the development of pharyngocutaneous fistula, constituted the most common postoperative complication in these patients. Multivariate analysis showed that primary CTRT was an independent predictor of local wound complications and pharyngocutaneous fistula. PMID- 15816050 TI - High-grade tumor differentiation is an indicator of poor prognosis in African Americans with colonic adenocarcinomas. AB - BACKGROUND: To identify the factors that contribute to poorer colon carcinoma survival rates for African Americans compared with Caucasians, the authors evaluated survival differences based on the histologic grade (differentiation) of the tumor. METHODS: All 169 African Americans and 229 randomly selected non Hispanic Caucasians who underwent surgery during 1981-1993 for first primary sporadic colon carcinoma at the University of Alabama at Birmingham or its affiliated Veterans Affairs hospital were included in the current study. None of these patients received presurgery or postsurgery therapies. Recently, the authors reported an increased risk of colon carcinoma death for African Americans in this patient population, after adjustment for stage and other clinicodemographic features. The authors generated Kaplan-Meier survival probabilities according to race and tumor differentiation and multivariate Cox proportional hazards models to estimate hazard ratios (HR) with 95% confidence intervals (95% CI). RESULTS: There were no differences in the distribution of pathologic tumor stage between racial groups after stratifying by histologic tumor grade. Among patients with high-grade tumors, 54% of African Americans and 21% of Caucasians died within the first year after surgery (P = 0.007). African Americans with high-grade tumors were 3 times (HR = 3.05; 95% CI, 1.32-7.05) more likely to die of colon carcinoma within 5 years postsurgery, compared with Caucasians with high-grade tumors. There were no survival differences by race among patients with low-grade tumors. CONCLUSIONS: These findings suggested that poorer survival among African-American patients with adenocarcinomas of the colon may not be attributable to an advanced pathologic stage of disease at diagnosis, but instead may be due to aggressive biologic features like high tumor grades. PMID- 15816051 TI - Perineural spread of cutaneous malignancy to the brain: a review of the literature and five patients treated with stereotactic radiotherapy. AB - BACKGROUND: The retrospective analysis was performed to investigate the role of stereotactic radiotherapy (SRT) techniques for patients with intracranial perineural spread (PNS) of a primary cutaneous malignancy. METHODS: Five patients were identified who received SRT from 1993 to 2003 for cutaneous malignancies with intracranial PNS to the cavernous sinus (n = 3) or Meckel's cave (n = 2). Patients were treated with GammaKnife stereotactic radiosurgery (n = 2), linear accelerator (linac)-based fractionated SRT (n = 2), or linac-based stereotactic radiosurgery (n = 1). RESULTS: The median overall survival (OS) periods from diagnoses of cutaneous malignancy and intracranial PNS were 63.0 months (range, 22.0-102.2 months) and 25.5 months (range, 22.0-55.2 months), respectively. The median OS from SRT was 24.2 months (range, 19.5-53.2 months). One patient was alive and without evidence of disease at 53 months of follow-up. The median durations of local and regional control from SRT were 19.5 months (range, 1.5 53.2 months) and 7.0 months (range, 1.5-53.2 months), respectively. CONCLUSIONS: Previous reports generally have recommended that patients with intracranial PNS receive palliative external-beam radiotherapy. Results from the current study suggest that some of these patients may have prolonged survival, or even may be cured. Judicious use of SRT should be considered in their management. PMID- 15816053 TI - Depressive symptoms among women with an abnormal mammogram. AB - An abnormal mammography finding constitutes a stressful event that may increase vulnerability by developing or intensifying pre-existing psychological morbidity. We evaluated depressive symptoms using the Composite International Diagnostic Interview among women of four ethnic groups who had an abnormal mammography result controlling for the effect of demographic, psychosocial and medical factors on recent onset of depressive symptoms. Telephone surveys were conducted among women aged 40-80 years recruited from four clinical sites in the San Francisco Bay Area after receiving a screening mammography result that was classified as abnormal but probably benign, suspicious or highly suspicious, or indeterminate using standard criteria. Among the 910 women who completed the interview, mean age was 56 (S.D.=10), 42% were White, 19% Latina, 25% African American, and 14% Asian. Prevalence of lifetime depressive symptoms was 44%, and 11% of women had symptoms in the previous month. Multivariate logistic regression models showed that Asian ethnicity, annual income >$10 000 and weekly attendance at religious services were significantly associated with decreased depressive symptoms. Having an indeterminate result on mammography and being on disability were significantly associated with more depressive symptoms. Reporting a first episode of depression more than a year before the interview was associated with significant increase in depressive symptoms in the month prior to the interview regardless of mammography result. Women with an indeterminate interpretation on mammography were at greater risk of depressive episode in the month prior to the interview compared to women with probably benign results (odds ratio=2.41; 95% CI=1.09-5.31) or with a suspicious finding. Clinicians need to consider depression as a possible consequence after an abnormal mammography result. PMID- 15816055 TI - Primary angioplasty should be performed in hospitals without on-site surgery. PMID- 15816054 TI - Prognostic factors and treatment of patients with T-cell non-Hodgkin lymphoma: the M. D. Anderson Cancer Center experience. AB - BACKGROUND: T-cell non-Hodgkin lymphomas (T-NHL) are more aggressive and patients have a poorer prognosis compared with patients with the corresponding B-cell lymphomas. Although intensive treatments have been developed, it is unknown whether they are more effective than CHOP chemotherapy (cyclophosphamide, doxorubicin, oncovorin, and prednisone). METHODS: The authors' retrospective study evaluated the clinical outcome of 135 previously untreated patients with T NHL who were treated at The University of Texas M. D. Anderson Cancer Center (Houston, TX) between 1996 and 2002. Lymphomas with T-cell histologies with the exception of mycosis fungoides were included. RESULTS: The estimated median overall survival was 46 months. Thirty-seven percent of the patients received CHOP therapy, 48% received intensive therapy, and 15% received other therapy. The estimated 3-year overall survival rates were 62% for the patients treated with CHOP therapy and 56% for the patients who received intensive therapy. After the exclusion of patients with anaplastic large cell lymphoma (ALCL), who are known to have a better prognosis than patients with other T-NHLs, the estimated 3-year overall survival rates were 43% for the patients treated with CHOP therapy and 49% for the patients who received intensive therapy. Parameters that may be independent prognostic factors for survival in T-NHL, excluding ALCL, included ECOG performance status > or = 2, beta-2-microglobulin level > 2 mg/L, lactate dehydrogenase level higher than normal, bulky disease > or = 7 cm, and a higher international prognostic index and tumor score. CONCLUSIONS: The current study data suggested that patients treated with intensive therapies did not fare better than those treated with CHOP therapy. New treatment regimens need to be developed for patients with T-NHL. PMID- 15816056 TI - Why we should not do percutaneous coronary intervention at sites without surgical backup. PMID- 15816058 TI - Patterns of comorbidity in panic disorder and major depression: findings from a nonreferred sample. AB - Previous findings in referred adult samples document major depression as having important moderating effects on the patterns of comorbidity for panic disorder and major depression. This study evaluated whether these patterns of comorbidity are moderated by referral bias. Panic disorder (PD) and major depression (MD) were used to predict the risk for comorbid psychiatric disorders and functional outcomes using data from a large sample of adults who had not been ascertained on the basis of clinical referral (N=1,031). Participants were comprehensively assessed with structured diagnostic interview methodology to evaluate childhood and adult comorbid psychiatric disorders. PD increased the risk for anxiety disorders, independently of MD. MD increased the risk for mania, antisocial personality disorder, psychoactive substance use disorder, disruptive behavior disorders, overanxious disorder, social phobia, and generalized anxiety disorder, independently of PD. These results extend to nonreferred samples' previously reported findings documenting that MD has important moderating effects on patterns of comorbidity for PD and indicate that patterns of comorbidity for PD are not due to referral bias. PMID- 15816061 TI - Candidate gene analysis in human neural tube defects. AB - Biochemical and developmental pathways, mouse models, and positional evidence have provided numerous candidate genes for the study of human neural tube defects. In a survey of 80 studies on 38 candidate genes, few found significant results in human populations through case-control or family-based association studies. While the folate pathway has been explored extensively, only the MTHFR 677C > T polymorphism was significant, and only in an Irish population. Developmental pathways such as the Wnt signaling pathway and Hox genes have also been explored without positive results. More than 90 mouse candidates have been identified through spontaneous and knockout mutations, but only the T locus (mouse Brachyury gene) showed association in an initial study that was not confirmed on follow-up. Positional candidates have been derived from cytogenetic evidence, but preliminary genomic screens have limited power due to small sample sizes. Future studies would increase their power to detect association by using more samples. In addition a clarification of the phenotype would be beneficial as many studies used different inclusion criteria. Incorporating several types of data could highlight better candidates, as would looking beyond the traditional sources for candidate genes. Recent studies of an energy metabolism gene (UCP2) and vitamin B metabolism (Transcoalbumin) have produced promising results. Utilizing other model organisms may also be beneficial, as in a recent study from a chick model of NTDs in NCAM1. New approaches combined with traditional methods and increased sample sizes will help prioritize human NTD candidate genes and clarify the complex etiology of this condition. PMID- 15816059 TI - Social anxiety disorder and depression in Saudi Arabia. AB - The purpose of this study was to estimate the prevalence of depression in patients with social anxiety disorder (SAD) and to assess the relationship between the severity of SAD symptoms and depression. Ninety-eight consecutive patients with generalized SAD according to DSM-IV criteria were included in a cross-sectional case-control study. Patients were referred to a psychiatric outpatient clinic in a general hospital in Saudi Arabia. The Liebowitz Social Anxiety Scale was used to estimate the severity of SAD. Fifty-eight (59%) of the patients with SAD had another current psychiatric disorder. Forty (41%) patients had current depression, and 37 (92.5%) of them had it after SAD onset. Eleven of 16 patients with severe SAD (69%) had depression whereas only 29 of 82 of SAD patients with mild or moderate subtypes (35%) had depression. Patients with severe SAD were four times more likely to have depression than the patients with mild or moderate SAD even after controlling for confounding sociodemographic and clinical factors. Depression is common among patients with SAD, particularly the severe subtype. Early recognition and treatment of SAD, especially the severe subtype, may prevent the occurrence of depression. Prospective studies are needed to investigate the risk factors that may lead to depression in SAD. PMID- 15816062 TI - Determination of protein-ligand binding affinity by NMR: observations from serum albumin model systems. AB - The usefulness of bovine serum albumin (BSA) as a model protein for testing NMR methods for the study of protein-ligand interactions is discussed. Isothermal titration calorimetry established the binding affinity and stoichiometry of the specific binding site for L-tryptophan, D-tryptophan, naproxen, ibuprofen, salicylic acid and warfarin. The binding affinities of the same ligands determined by NMR methods are universally weaker (larger KD). This is because the NMR methods are susceptible to interference from additional non-specific binding. The L-tryptophan-BSA and naproxen-BSA systems were the best behaved model systems. PMID- 15816063 TI - UK: House of Lords recognizes that privacy rights include protection of mental health. AB - Razgar, an Iraqi national, had claimed asylum in the United Kingdom. He was not HIV-positive. He suffered from a psychiatric disability and was challenging an order that he be removed to a third country on the basis that it would detrimentally affect his mental health. PMID- 15816064 TI - A drug may help hot flashes after prostate cancer treatment. PMID- 15816065 TI - Meditation. Tuning the mind to help heal the body. PMID- 15816066 TI - I saw part of a news report linking heartburn medication to pneumonia. Please explain. PMID- 15816068 TI - Heavy hula hoop for your health? PMID- 15816067 TI - Is canned salmon I see at the supermarket as good for you as fresh salmon? PMID- 15816069 TI - MRSA: how politicians are missing the point. PMID- 15816070 TI - Reining in direct-to-customer advertising. PMID- 15816071 TI - Metamphetamine use linked to rising HIV transmission. PMID- 15816072 TI - Russian scientists voice concern over "stem-cell cosmetics". PMID- 15816073 TI - Chlorhexidine-modified glass ionomer for band cementation? An in vitro study. PMID- 15816075 TI - Looking into heart disease. Changes inside the eye can reflect the damage of high blood pressure and help gauge the risk of a future stroke or heart attack. PMID- 15816074 TI - Paul E. Lacy. PMID- 15816076 TI - Carnitine promising, but no miracle for heart disease. Hype for the dietary supplement carnitine as a treatment for heart disease far outstrips the evidence. PMID- 15816077 TI - Pain might mean gain for aneurysm repair. Surgery to repair an abdominal aortic aneurysm is harder on the body than an inside-the-aorta approach, but it's also more durable. PMID- 15816078 TI - Who needs Plavix? This super aspirin is meant mainly for people at high risk of having a heart attack or stroke. PMID- 15816079 TI - Beta testing. PMID- 15816080 TI - Ask the doctor. Sometime soon I will need to have my aortic valve replaced. I like the fact that mechanical valves last longer than biological valves, but I don't like the fact that they require you to take the blood thinner Coumadin to prevent blood clots. I have heard that one of the new mechanical valves won't require Coumadin, just aspirin. Is that so? PMID- 15816081 TI - "Profiling" breast cancer tumors could lead to individualized treatment. PMID- 15816082 TI - Statins do more than lower cholesterol. These powerful drugs may also protect the heart by reducing inflammation. PMID- 15816083 TI - Asthma in women after menopause. Hormone replacement therapy may explain why many women over 50 develop this disease. PMID- 15816084 TI - Dual-chamber pacemakers worth their cost. PMID- 15816085 TI - Heed early warning signs of ovarian cancer. Abdominal pain and changes in urinary continence could be indicators. PMID- 15816086 TI - Vitamin E from select plant seeds could inhibit lung and prostate cancer growth. PMID- 15816087 TI - Maintaining fitness keeps you sharp in old age. Men who decrease their physical activity after age 70 have greater mental decline than men who stay active. PMID- 15816088 TI - Training helps people with Parkinson's avoid falls. PMID- 15816089 TI - Standard treatment for chronic pain may pose greater risk. Along with the potential to cause ulcers and bleeding, NSAIDs could damage the small intestine. PMID- 15816090 TI - Pelvic floor exercise can reduce stress incontinence. PMID- 15816091 TI - Banishing ulcer-causing bacteria gives rise to appetite hormone. PMID- 15816092 TI - Warding off macular degeneration. The case for vitamin and mineral supplements to fight AMD is getting stronger with each study; but so is the case for good nutrition. PMID- 15816093 TI - Spinal surgery treats osteoporosis. Cementing spinal fractures stops crushed bone fragments from rubbing painfully against nerve endings. PMID- 15816094 TI - Weekly osteoporosis treatments also problematic. PMID- 15816095 TI - I had atrial fibrillation for a long time. My cardiologist recently tried me on amiodarone, and I am finally in a normal heart rhythm. Are there any side effects from this medication? PMID- 15816096 TI - University research ethics committees: their role, remit and conduct. PMID- 15816097 TI - Notes from an ethics fellowship. PMID- 15816098 TI - Ethical and legal issues in caring for asylum seekers and refugees in the UK. AB - Inward migration to the UK remains topical and controversial as numbers continue to increase. Many immigrants have specific health care needs and may shoulder a large burden of infectious disease. Imposition of legal constraints can have a huge impact on the medical care afforded to immigrants. Currently UK policy is to treat, free of charge and with NHS resources, those who fulfil specific criteria. However an increasing number are being asked to pay for their treatment. Many health care professionals are confused as to current legal restrictions and require guidance on the associated ethical issues. We concentrate on provision of care to HIV positive individuals and use cases to illustrate some of the issues. However these issues are equally pertinent to practitioners in all branches of medicine. PMID- 15816099 TI - Role of the microbiology laboratory in infectious disease surveillance, alert and response. AB - Surveillance is usually defined as the ongoing and systematic collection, analysis and interpretation of health data essential to the planning, implementation and evaluation of public health practice. During recent years, most of these programmes have been developed in the field of antimicrobial resistance and nosocomial infections, but efforts have also been made in other areas. Recent experiences of emerging microbial threats, including severe acute respiratory syndrome and new influenza variants affecting humans, the re emergence of infectious disease problems and the possibility of bioterrorism have evidenced the need for implementation of infectious disease surveillance programmes. clinical microbiology laboratories play a pivotal role in these programmes. They have the first opportunity to detect these problems and should participate in the design of reporting strategies and dissemination of this information. Policies for the flow of data to national and international authorities should be established using passive surveillance strategies. However, active surveillance programmes taking advantage of new methodologies, including virtual tools and mathematical programs, should be the goal for early detection of unusual patterns of microbial pathogens, outbreaks and healthcare-associated infections. In addition, early implementation of response strategies should be designed and performed with the cooperation of microbiology laboratories, and intervention and response protocols should be defined with the participation of clinical microbiologists. PMID- 15816100 TI - Infectious disease emergencies: role of the infectious disease specialist. AB - The importance of infections for public health has become obvious during the last decades. Examples are emerging infections such as HIV/AIDS and severe acute respiratory syndrome, deliberate release of microorganisms, such as the anthrax episode in the USA, the increasing problems with organisms resistant to antimicrobial treatment, such as methicillin-resistant Staphylococcus aureus, and the threat of a new influenza pandemic with a case fatality rate similar to that in the 1918 outbreak. An effective response to infectious disease emergencies requires careful planning and establishment of resources in advance. The medical specialties involved are clinical microbiology, clinical infectious diseases and epidemiology. Clinical microbiology should include bacteriology, virology, and parasitology; the technical developments during the last 15 years have clearly erased most of the methodological differences between these branches of microbiology. New techniques such as new generations of Polymerase Chain Reaction (PCR), rapid methods for nucleic acid sequence analyses and microarrays have enabled more rapid identification of organisms and provide powerful tools in the epidemiological analysis of an outbreak. The infectious disease specialists are necessary for rapid and adequate clinical diagnoses, optimal use of antimicrobial agents and provision of facilities for containment of patients who may spread the infections. The need for isolation units became acute when many countries prepared themselves for a possible severe acute respiratory syndrome outbreak in Europe. With few exceptions, Europe still lacks epidemiological field forces, and it has been embarrassing to be obliged to call upon the Centers for Disease Control for European outbreaks. Hopefully, this will be corrected with the creation of the European Centre for Disease Prevention and Control (ECDC). PMID- 15816101 TI - The ethical governance and regulation of student projects: a draft proposal. PMID- 15816102 TI - Changing needs, opportunities and constraints for the 21st century microbiology laboratory. AB - Clinical microbiologists and microbiology laboratories are experiencing changes due to evolving views on 'healthcare delivery' as an economic activity, due to changes in the medical environment and the demographics of the workforce, and technical evolution. Cost-effectiveness of laboratory procedures has been achieved through consolidation and integration of laboratories. Consolidation offers economy of scale and reduction in numbers of on-site staff, but also leads to separation of microbiologists from their clinical colleagues. Integration puts different laboratory disciplines under a single management, and leads to reorganisation of laboratories along common work-lines. Cost-savings combined with on-site availability of laboratories are achieved at the expense of a reduction in the influence of microbiologists in the daily running of the laboratory. Medically, there is growing emphasis on evidence-based diagnostics. Because of time-delays inherent in culturing, microbiology through rapid testing is mandatory. There is an increasing shortage in Europe and the USA of trained microbiology laboratory technicians and microbiologists. This reinforces the trend towards more automation and integration. Technological advances, particularly in molecular diagnostics, offer the possibility of rapid reporting and improvement of the impact of clinical microbiology on patient management. Molecular tests, however, fit perfectly the concept of an integrated laboratory and may further loosen the link between microbiologist and microbiology tests. The challenge for clinical microbiology will be to use new techniques to improve its cost-effectiveness and impact on infectious disease management. The future organisation of microbiology laboratories must support this but is itself of secondary importance. The training of future microbiologist must prepare them for this changing environment. PMID- 15816103 TI - Continuing professional development and clinical governance: the role of scientific societies. AB - To date, the rules and state of development of the processes of continuing education and the evaluation of skills and competence vary considerably from one European Member State to another. The recognition of the freedom of establishment of health professionals throughout Europe must be made conditional upon the possession of a given qualification, and also the demonstration of maintained level of expertise, knowledge and skills. This appears to be of primary importance in order to maintain a good quality of care and to improve the performances and responsibilities of the infection of specialists, within the healthcare system. The role of scientific societies such as the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) can be envisaged as follows: (1) to play a key role in the coordination of the processes; definition of the main topics and visions, accreditation of the teaching courses and modalites of evaluation (which supposes a high level of cooperation with the platform for professional qualification) (2) to manage training courses (e.g., ESCMID School, postgraduate courses, technical workshops, educational activities within congresses). In order to make the system clearer and easier to apply, a proposal for a single, comprehensive directive is highly necessary. PMID- 15816104 TI - Proceedings and abstracts of the National Conference on Elimination of Leprosy. Raipur, India, 27-30 January 2004. PMID- 15816105 TI - Design of artificial environments on the basis on human sensibility. Proceedings of the International Symposium on Design of Artificial Environments. 2004. Fukuoka, Japan. PMID- 15816106 TI - The influence of epidermal growth factor receptor and tumor differentiation on the response to accelerated radiotherapy of squamous cell carcinomas of the head and neck in the randomized DAHANCA 6 and 7 study. AB - BACKGROUND AND PURPOSE: Reduction of the overall treatment time of radiotherapy has increased locoregional control and disease specific survival in squamous cell carcinomas of the head and neck (HNSCC), but the response is heterogeneous. EGFr is often overexpressed in HNSCC and has been related to the repopulation taking place during radiotherapy. The aim of the current study was to address the influence of EGFr and histopathological differentiation when the overall treatment time of radiotherapy was moderately reduced. PATIENTS AND METHODS: Eight hundred and three patients with representative pretreatment tissue samples from the randomized DAHANCA 6 and 7 study of 5 vs. 6 fx/wk of radiotherapy. EGFr was visualized using immunohistochemistry and separated into high and low expression before correlation with clinical data. RESULTS: Tumors with high EGFr (84%) responded better to moderately accelerated radiotherapy, than carcinomas with low EGFr, using locoregional control as endpoint and a similar pattern was seen, stratifying by well/moderate vs. poor tumor differentiation. Therefore, a combined parameter was constructed showing a more prominent separation of response: tumors with high EGFr and well/moderate differentiation did benefit from moderate acceleration of treatment regarding locoregional control, HR 0.54 (0.37-0.78), whereas such an effect was not seen in tumors with low EGFr and/or poor differentiation, HR 0.8 (0.51-1.25). These results reflected the disease specific survival as well and were confirmed in multivariable analyses. CONCLUSIONS: Moderately accelerated fractionation is superior to conventional treatment in HNSCC but the response is heterogeneous and may be predicted by high expression of EGFr and well/moderate tumor differentiation. PMID- 15816107 TI - The expression of epidermal growth factor receptor results in a worse prognosis for patients with rectal cancer treated with preoperative radiotherapy: a multicenter, retrospective analysis. AB - BACKGROUND AND PURPOSE: Expression of epidermal growth factor receptor (EGFR) is observed in 50-70% of colorectal carcinoma and is associated with poor prognosis. The aim of this study was to determine the prognostic value of EGFR status before radiotherapy in a group of patients with locally advanced rectal cancer treated with preoperative radiotherapy. PATIENTS AND METHODS: Eighty-seven patients were studied retrospectively. Treatment consisted of pelvic radiotherapy, in 50 patients with concomitant chemotherapy and surgical resection. Immunohistochemistry for EGFR was determined at the preradiation biopsy and in the resected specimens. Immunohistochemical analysis for EGFR expression was evaluated according to extension and staining intensity. We defined positive staining (EGFR positive), when extension was 5% or more. RESULTS: A total of 52 of 87 tumors showed EGFR positive status at biopsy (60%) and EGFR expression was associated neither with clinical tumor stage nor with clinical nodal stage. EGFR positive expression was linked to a lack of pathologic complete response to preoperative radiotherapy (P=0.006). Disease-free survival was lower among patients with EGFR positive status before radiotherapy (P=0.003). In a multivariate analysis EGFR expression at biopsy was a statistically significant predictor of disease-free survival, RR=2.88(1.1-7.8), P=0.036. CONCLUSIONS: EGFR is expressed in a significant number of rectal tumors. EGFR-positive expression before radiotherapy is an indicator for poor response and low disease-free survival. PMID- 15816108 TI - Clinical epidemiology of transfusion medicine. PMID- 15816109 TI - SAMHSA releases revised and updated treatment improvement protocol on co occurring disorders. PMID- 15816110 TI - [Academician of the Russian Academy of Medical Sciences Fedor Grigor'evich Uglov, on his 100th birthday and 75th year of his surgical activities]. PMID- 15816111 TI - Development of a short form inventory to assess grief in caregivers of dementia patients. AB - This article describes the derivation of a short-form of the Marwit-Meuser Caregiver Grief Inventory (MM-CGI), an inventory designed to measure grief in caregivers of persons with progressive dementia. It presents initial reliability and validity data and describes ways to use the inventory both clinically and scientifically. The resulting MM-CGI Short Form, presented here in its entirety, is comprised of 18 items and, like its full-form relative, is self-scoring and yields a Total Grief score and sub-scale scores measuring factors representing Personal Sacrifice Burden, Heartfelt Sadness and Longing, and Worry and Felt Isolation. PMID- 15816112 TI - Responding to suicidal calls: does trait anxiety hinder or help? AB - To see if trait anxiety and suicidality interfered with the ability to respond to suicidal crisis calls, 279 undergraduates completed measures of trait anxiety and suicidality in the past week, and the revised Suicide Intervention Response Inventory (SIRI-2). Unexpectedly, trait anxiety (but not suicidality) correlated with better SIRI-2 scores. In addition, students scored significantly more like the experts on the SIRI-2 if they were European-American, were relatively older, had completed a crisis intervention course, had undergone psychotherapy, or had ever been suicidal in their lives. Trait anxiety does not hinder, and may even help, in responding to a suicidal phone call. PMID- 15816113 TI - Narratives of grieving African-Americans about racism in the lives of deceased family members. AB - As part of a comprehensive interview study on African-American grief, the authors explored how racism is incorporated into narratives about a deceased family member. To the extent that experiences of racism are pervasive in African American life and to the extent that narratives about a person who has died generally account for the life experiences, achievements, character, and challenges faced by the deceased, the authors expected narratives about a deceased African-American to deal with the person's encounters with racism. In fact, most of the 26 African-Americans who were interviewed spoke about racism in the life of the deceased. Many talked about racism blocking the deceased from getting ahead occupationally and in other ways and about how the deceased resisted or stood up to racism. Some people spoke about the ways the deceased taught them to deal with racism. The narratives that indicated that the deceased had rarely or ever talked about racism still made clear that the deceased lived in a racist world but chose not to bring racism into family conversation. The findings suggest that a view of African-American grieving that is insensitive to racism in African-American experience may lead to unhelpful grief support or counseling. PMID- 15816114 TI - Childhood mourning: prospective case analysis of multiple losses. AB - Multiple losses within short time periods make one question life and can exponentially influence one's coping skills. But what are the effects on a child and what should be done when the next loss occurs? This case addresses the multiple losses suffered by a child while assessing coping skills of the child and coping strategies used by the parents to assist the child. PMID- 15816116 TI - The future of pathology as a discipline: none without a dictionary! PMID- 15816117 TI - [ What is your diagnosis? Answer: Supratentorial primitive neuroectodermal tumor (sPNET), G4]. PMID- 15816118 TI - [Acute aortic syndromes]. AB - The term acute aortic syndrome comprises in addition to classic dissection also two recently described entities--penetrating ulcer and intramural haematoma. Forty-two necropsy cases of acute aortic syndrome were encountered during the last six years--41 dissections and one penetrating ulcer; the intramural haematoma was not seen. According to the DeBakey classification, there were 26 dissections of type I, 8 of type II, 1 of type IIIA, and 4 of type IIIB; in 2 cases the dissection was confined to the abdominal aorta. The dissection had a chronic character in four cases. Aortic rupture was found in 24 cases (59%), most frequently (19x) into the pericardial cavity. The aortic dissection continued into aortic branches in 25 patients (61%), causing stenosis/obstruction of coronary arteries in 7, of branches of the arch in 20, of abdominal arteries in 12, and of renal arteries in 17 patients, respectively. Histologically, there were degenerative lesions of the Erdheim type in the media of 10 aortas (from 21 completely examined). As possible risk factors for aortic dissection there appeared hypertension in 32 patients, anuloaortic ectasia in 11, saccular aneurysm ot the abdominal aorta in 6, family history of dissection in 2, Marfan syndrome in 2, prolaps of the mitral valve in 2, and bicuspid aortic valve in 2 patients, respectively. The aortic dissection was iatrogenic in 7 patients, presenting as a complication of a cardiosurgical or invasive cardiological procedure. PMID- 15816119 TI - [The role of the extracellular space in biology of glial brain tumors]. AB - The size, geometry and composition of the extracellular space (ECS) play an important role in influencing the biological behavior of primary brain tumors. Experiments employing the real-time TMA iontophoretic method to determine the size and geometry of the ECS, by monitoring the diffusion of TMA ions in the ECS, revealed a dramatic increase in ECS size in brain neoplasms when compared with that of unaffected brain cortex. Further, the increase of ECS volume in tumors was shown to correlate with increasing proliferative activity and increasing cellularity of astrocytomas. The increase in ECS size was surprisingly accompanied by a significant increase in diffusion barriers, slowing the diffusion of molecules in the ECS of tumors. In low-grade tumors, diffusion is hindered by the presence of a dense net of tumor cell processes. In high-grade gliomas, in which the cellular processes are shortened with reduced branching, the increase in diffusion barriers is caused by the overproduction of specific components of the extracellular matrix (ECM) by the tumor cells, mainly tenascin. The ECM glycoproteins produced represent a substrate for the subsequent adhesion and migration of tumor cells through the enlarged ECS. However, they might also critically reduce the diffusion of therapeutics into the tumor. The presence of tenascin in the ECS of a neoplasm correlates significantly with the increased malignancy of the tumor and a poor clinical outcome of the disease, thus making the immunohistochemical detection of tenascin diagnostically useful as a prognostic marker and a marker of aggressive biological behavior of tumors. PMID- 15816120 TI - [Nervous component of mature cystic ovarian teratomas]. AB - In 30-50 percent of cases mature cystic ovarian teratomas contain a nervous tissue which can be highly differentiated. This study was focused on research of the nervous component of mature cystic ovarian teratomas with generally available methods to pathologists, including impregnation techniques, immunohistochemistry and electron microscopy. From the total number of 212 mature cystic ovarian teratomas, the nervous tissue was found in 72 cases (34%), which corresponds to the literature data. According to its differentiation, it was possible to distinguish five categories of nervous tissue by light microscopy: 0 peripheral nervous tissue only, 1--solid glial nodules, 2--glial cysts, 3--glial tissue with abundant scattered neurons and, finally, 4--organoid nervous tissue similar to certain CNS structures. Apart from the foci similar to grey matter of the spinal cord and cerebral cortex, those of differentiated cerebellar cortex were present as well. Astrocytes mostly predominated in the nervous tissue, and they sometimes showed reactive changes including gemistocytes and formation of Rosenthal fibres. Neuronal elements also showed degenerative changes quite frequently, especially in a less differentiated nervous component. These changes might have developed due to an abnormal location of the nervous tissue or its hypoxia in the teratoma. Contrary to some literature data, oligodendrocytes and myelin were present in the nervous tissue of most of our cases. Ultrastructurally, neurons with fully developed synapses were observed in the nervous component, and dendritic spines were present on dendrites of Purkinje cells of cerebellar cortex. The results obtained from the examination of teratomas in this study confirmed and enriched the literature data concerning the high degree of differentiation of their nervous components. We suggest that the differentiated nervous tissue of teratoma represents a unique natural model suitable for research of some aspects of neurohistology and neuropathology, e.g. synaptogenesis or myelinogenesis. PMID- 15816121 TI - [Acinic cell-like differentiation in invasive ductal carcinoma and in ductal hyperplasia of the breast--report of two cases]. AB - Described are two epithelial lesions of the breast displaying extremely rare, widespread acinic cell-like differentiation (metaplasia). Two women, 70 and 40 year-old, one with invasive ductal papillocarcinoma, the other one with conventional intraductal hyperplasia without atypia, both demonstrated massive diffuse, PAS positive, granular eosinophilic transformation of the cell cytoplasm. This unusual cell appearance closely simulated acinar cells in normal serous salivary gland/acinic cell carcinoma or Paneth cells. Both extensive expression of lysozyme and finding of numerous zymogen granules ultrastructurally confirmed the acinic cell-like fenotype. Discussed is differential diagnosis of the breast neoplasm containing overt eosinophilic and granular cytoplasm. Reviewing literature and comparing our unique finding of unusual salivary-type differentiation in conventional ductal hyperplasia of the breast, biologic implications are considered. PMID- 15816122 TI - [Anaplastic carcinoma of the thyroid gland with chondrosarcomatous component]. AB - The case of an 83-year-old female with a history of a two months enlarging goiter is presented. Clinically, a firm goiter with fixation to surrounding soft tissues was found. Fine needle aspiration cytology was performed with the conclusion of a sarcomatoid variant of anaplastic carcinoma of the thyroid gland; biopsy was recommended. The patient was operated on and died six months after operation with metastatic dissemination in lymphatic nodes and lungs. Histologically, an invasive high-grade tumor composed of irregular, sometimes bizarre spindled or polygonal cells intermingled with foci of chondroid tissue was seen. The cartilage comprised lobularly arranged chondrocytes with irregular nuclei; bi- or multinucleated chondrocytes were present as well. The diagnosis of thyroid anaplastic carcinoma with chondrosarcomatous component was established. Differential diagnostics of this tumour is discussed. PMID- 15816125 TI - [General situation, stalemate and future]. PMID- 15816123 TI - [Humor from the mortuary--patho-anatomic historic anecdotes--part 1]. PMID- 15816126 TI - [Prolegomena to reform of healthcare services: from identification of problems to development of effective tools]. AB - Various institutions have developed healthcare programs to cope with the serious epidemiologic situation in many developing counties. Although these programs are indispensable, many are "disconnected" from the social context for which they were developed. Perusal of a few anthropological texts describing problems associated with providing healthcare services in Africa allows identification of the difficulties and modalities of interaction between caregivers and populations and a basis for proposing avenues for reflection about improvement. PMID- 15816127 TI - [Results of primary healthcare]. AB - The concept of primary healthcare was formulated at the international conference held in Alma-Ata (USSR) on September 6 to 12, 1978. Over the past quarter century, the term "primary healthcare" has been widely used as a basis for implementing healthcare policies in various nations, especially those considered as developing countries. However many programs initiated in the name of the primary healthcare concept have only partially complied with the spirit of the final declaration of Alma-Ata. Analysis of the healthcare in Mali shows that the system followed that evolution. Although considerable time and energy were regrettably wasted in implementing the village health agent strategy, this phase may have been a necessary step in the political development of the state at that time. Recent changes in the country's healthcare system in compliance with the original principles of primary healthcare show that the 1978 declaration is sound and remains pertinent as democracy makes further gains in the world. PMID- 15816128 TI - [For front-line community-based general practitioners]. AB - In the past thirty years considerable effort has been devoted to delivering basic health services to rural populations. However the quality of care remains poor. One problem is that the absence of general practitioners working at the community level has resulted in health care systems without doctors. This "missing-link" situation is inconsistent with the availability of a large number of young fully trained medical doctors who graduated from Medical Schools in West African countries and Madagascar. Based on examples in Mali and Madagascar, the authors of this article attempt to show the advantages of a new concept, i.e., community based general practitioners. These practitioners would not only provide a front line medical presence but also combine the principles of primary health care (PHC) and family medicine (FM) into a single practice. For this approach to be successful a number of obstacles would have to be overcome and several indispensable accompanying measures would have to be implemented. The goal would be to establish a familiar front-line healthcare provider that village communities could trust. PMID- 15816129 TI - [Public hospital in French-speaking Africa]. AB - Traditional consensus holds that hospitals are ill-suited to the healthcare priorities in Africa countries whose policies must focused mainly on promoting primary services. Indeed hospitals are generally considered as inefficient and wasteful of financial resources that could be used for more important healthcare priorities. Long excluded from major development programs, most hospital facilities have gone from crisis to crisis over the last 10 years and are now unable to provide adequate services. The sometimes dramatic state of these institutions raises serious doubt not only about the effectiveness of current healthcare policies but also about the long-term survival of the healthcare systems now being established. But this situation is not the result of fate but rather of the failure of public hospitals to adapt their organization to the realities of today's world. Indeed hospital services are provided within a purely administrative structure with total disregard for the principles of good management. A new vision for revival of healthcare systems make hospitals a key component in a network of healthcare institutions and recommends that they be included in an in-depth reform of healthcare systems that consider any healthcare act as a service in the "economic" sense of the word and its availability as the end-result of a production process. To ensure fulfillment of the public service mission of the hospital, the resulting "entrepreneurial" approach must be accompanied by appropriation of subsidies so that charges are affordable to a majority of the population, implementation of welfare programs to insure that even the poorest users have sufficient resources, and development of monitoring capabilities. PMID- 15816130 TI - [Financing healthcare in low-income countries: recurring questions, new challenges]. AB - Healthcare financing policies in low-income countries have gone through three successive phases. In the first phase the dominant approach was based on free access to healthcare and focused first on development of vertical programs and then on the necessity of providing primary care to all. While maintaining the emphasis on accessibility to primary care, the second policy phase introduced user fees and attempted to integrate healthcare programs into district-based healthcare structures. The third phase has been strongly influenced by the relationship between healthcare and development and the Millenium Objectives and places strong emphasis on necessity of developing insurance schemes. Recent studies on the relationship between healthcare spending and health status indicate that the efficiency and effectiveness of healthcare spending plays a more determinant role than the amount. At the same time an effort is being made to develop synergy between the different players in the health care systems and to clarify the role of each player by hinging financing decisions on operating criteria such as "public welfare", externalities, catastrophic costs, and equity. Although many countries have made significant progress, there are still several lagging areas, i.e., coverage for the poorest segment of the population (despite the rhetoric), follow-up of financing, and governance. Increasing external aid already initiated by several states may have a non-negligible impact on the macroeconomic balance. Since these changes could lead to adverse effects on health, there is a need to implement careful non-dogmatic policies. PMID- 15816131 TI - [Healthcare: a growing role in international politics]. AB - Since the end of the cold war the tone of international relations has clearly changed. Whereas relations were once defined strictly in terms of more or less armed confrontation, economic and social issues now play a growing role. Healthcare policies in Africa have long been influenced by the policies of countries sponsoring bilateral and even multilateral foreign aid programs. However the last ten years have witnessed an increasing interaction between international policy and healthcare policy. The two main reasons for this trend involve 1) access to drug treatment and the WTO and 2) the extension and impact of the AIDS epidemic. The problem of access to drug treatment for poor populations (fundamental right) has led to the emergence of an increasingly strong and effective civil society. Because of its social and economic effects as well as its geopolitical and security implications, AIDS has become a major factor in international relations. With regard to both these issues the place and role of the USA is demonstrative of the interaction between healthcare and international relations. PMID- 15816132 TI - [Healthcare in African cities: from preconceptions to recognition of a public health emergency]. PMID- 15816133 TI - [Reducing maternal mortality in developing countries: theory and practice]. AB - Chiefs of state attending the Millennium Summit (2000) set a goal of reducing maternal mortality by 75% before 2015. Based on knowledge of the epidemiology of maternal mortality/morbidity and on growing experience in the field, the international community defined a relatively low-cost program of evidence-based initiatives. However implementation of that program has been stymied by the reality that increasing geographical accessibility to a full range of quality emergency obstetric care of quality will require large investments of money and time. Increasing financial accessibility remains difficult given the low standard of living of populations and budget cutbacks by national governments. The problems facing women and health workers are mostly overlooked by public health policy. There is need for a multi-disciplinary approach with equal participation of specialists in public health, gyneco-obstetrics, anthropology, health care economics, political science and social and community mobilization. PMID- 15816134 TI - [Malaria at the time of "Roll Back Malaria"]. AB - Only within the last 4 years has worldwide political awareness of the impact of malaria made it possible to envision significant initiatives in malaria control and research within the framework of an ambitions and sustained program called the "Role Back Malaria (RBM) Partnership" coordinated by the WHO. The purpose of this article is to describe the current main objectives of the RBM partnership. This program implies a transfer of responsibilities and research technologies, which will require adaptation. Up to now French expertise has played little part. PMID- 15816135 TI - [The fight against AIDS in Africa: perspectives and responsibilites]. AB - As summarized in this article, key initiatives undertaken over the past 20 years have not succeeded in controlling the spread of AIDS in sub-Saharan Africa. Achievements are tenuous and local in scope. Two major handicaps have been dysfunctional national governments especially in the health sector and poor coordination of national and international strategies. As the epidemic spreads to Eastern Europe and Asia, there is an increasing need for effective application of the "three ONES principle", i.e., ONE action plan, ONE national body for coordinating, ONE monitoring and evaluation system. PMID- 15816136 TI - [Tuberculosis control in the world: results and challenges]. AB - The modern tuberculosis control strategy, which focuses on identifying and curing infectious cases, has made remarkable progress in recent years. This strategy, known as DOTS, receives significant support from bilateral and multilateral donors, in particular from the recently created Global Fund against AIDS, Tuberculosis and Malaria. Although case finding rates are still far too low everywhere in the world (45%), cure rates among infectious cases (target 85%) are progressing, but are still too low in Africa (71%). Nevertheless, the spread of the HIV/AIDS epidemic had led to a dramatic increase in the number of tuberculosis cases, even in countries where tuberculosis programmes have been functioning well for several years; in those countries that are heavily affected by HIV/AIDS, tuberculosis seems to be difficult to control if no progress is made in controlling the HIV epidemic. Health services are often too centralised, particularly in the big cities, and all health structures need to be involved in fighting tuberculosis. Treatment possibilities are limited; new research has been initiated to find new anti-tuberculosis drugs, but it is extremely important to take care to avoid the development of resistance to those drugs that are available. The main challenge in vanquishing tuberculosis is still that of development, mainly of the health services, and particularly their human resources. Tuberculosis elimination is necessarily a long-term goal that will require constant effort spanning several decades. PMID- 15816137 TI - [Public health training in subsaharan Africa: challenges and opportunities]. AB - Public health is a complex domain that covers a vast array of disciplines. Specialization in public health therefore requires a combination of various skills in equally complex fields. The public health training institutions in Africa are thus still unable to meet the huge need for adequate training in this domain. Counting solely on sending Africans to developed countries would neither quantitatively nor qualitatively fulfill the continent's enormous public health needs, either. To meet these needs, some African countries have created their own public health schools and offer local graduate training programs in various public health disciplines. A major advantage of these schools in comparison with the schools in developed countries is that the degrees they offer are immediately recognized by local African administrations. In addition, their training programs specifically focus on the existing public health needs and problems of African countries. Training Africans locally also contributes to reducing their temptation to emigrate after their graduation. However, the lack of adequate resources of African schools and their tendency to encourage premature high level specialization represent serious threats for the school's further development and long-term effectiveness. The lack of adequate management strategy to utilize well the trained cadres is still of great concern. It is along with poor wages and salaries among the key determining factors of the brain-drain. Poor management strategy appears also to be the root causes of the failure of the short term training programs. The effectiveness of these short term training programs is currently questionable because of their unnecessary huge numbers, and the focus on the payment of "perdiems" as their main benefit. Many obstacles to providing adequate local training to African public health providers remain to be overcome. However there are a number of opportunities including use of new information and communication technologies, partnerships between institutions, and use of modern management techniques. These opportunities can greatly improve the situation if they are properly used. PMID- 15816138 TI - [Experience with field epidemiology training programs in Africa: Cours IEIAA]. AB - The International Course on Epidemiology and Applied Data Processing in Africa (Cours IEIAA) is a practical 4-week French-language training course for professional health care personnel working in Africa. The first course was organized in 1998 by the Association pour l'Aide a la Medecine Preventive (Association for the Promotion of Preventive Medicine). The course organizing committee includes a pedagogical director and a faculty of field epidemiology experts. Course activities are designed to give attendees competency in epidemiology, biostatistics, computer techniques and communication. Attendance is funded mainly through education grants from the French Foreign Affairs Ministry, WHO, UNICEF, Belgian Technical Cooperation, Swiss Technical Cooperation, and German GTZ. Based on 7 years of experience, consideration is now being given to transforming this course into a tutorial Field Epidemiology Training Program modeled after those now operating in Ghana and Uganda [TEPHINET members Programs located in Africa]. PMID- 15816139 TI - [Issues on user participation in health care management in Africa]. AB - The 1990s witnessed great progress in increasing community participation in the management of health care services as an objective for reform of healthcare systems especially in urban areas benefiting from funding from the international community. Community participation has taken various forms from one country to another in terms of sources of healthcare training (public, private, or both), organization of management committees (inclusion or not of healthcare personnel), and scope of public service assigned to district health care units (preventive and/or curative care, management of proceeds from provision of health care procedures and/or medication, etc.). These strategies have had variable results and, although some urban programs have been evaluated, no attempt has been made to use this experience as a basis for analyzing the political implications of issues involving citizenship and public health. This report presents some ideas on these issues from the point of view of both governments and citizens and restates the purpose of user participation in healthcare services in Africa. That intent involves the need not only to increase household contributions to the cost of healthcare especially within the uncertain economic environment of urban areas but also to improve access to as well as quality of healthcare services. PMID- 15816140 TI - [The tropical pathogenic complex: a new look at an old concept]. AB - The failure of programs implemented to improve public health in the world, the poor performance of health care systems, and the difficulty in anticipating and reacting to emerging disease suggest that a more global approach to individual and community health problems is needed. Risk and disease must be correlated in space and time based on precise identification of their respective determinants, predictive factors, preventive targets, and diagnostic and prognostic indicators. For this purpose it could be useful to revive the pathogenic complex concept defined by the geographer Maximillien Sorre in 1933 in the light of new tools allowing better identification of biologic determinants (molecular genetics), of environmental factors (geographic information systems), and of behavior related to health care service activities. This concept could be used to place disease threats in a global context and to assess the consequences of current upheavals such as globalization of populations and products, exploding demographics, rapid urbanization, deforestations, rapid changes in behavior, and health care systems that can lead to both good and bad effects. PMID- 15816141 TI - [Control of tuberculosis as seen "in the field": popular perceptions of the disease and access to care in Conakry, Guinea]. AB - Tuberculosis is a public health priority especially in developing countries. A variety of control programs have been implemented. The purpose of this survey was to study the perceptions and practices of the victims of this disease "in the field". The findings of this survey carried out in a cohort of 65 patients and their families in Conakry, Guinea underline the importance of popular perceptions of the disease in care-seeking behavior. This survey also shows that health care workers share the same perceptions as the general population, which constitutes a major impediment for screening and treatment. PMID- 15816142 TI - [Communicating about risks to public health]. AB - The goals of public health risk communication are to explain the risks, to inform a community without causing a crisis, to implicate the population in decision making, and to improve dialogue between organizations and communities. Based on a document entitled "Communicating about risks to public health: pointers to good practice", the authors of this article review the main principles relevant to health risk communication and present a list of key points for effective management of this communication. PMID- 15816143 TI - [Is there a tropical specificity of public health? Post face attempts!]. AB - Public Health is not only a Medical speciality. It is a wide duty area, gathering together professional issued from various jobs. Worldwide is object is to implement, to manage and to assess policies aiming to better health for people. Since few decades these policies are mainly thought and imposed by national, sometimes international offices. Results are not these expected. In tropics, we must include anthropologic aspects and wide sociologic variability to adapt Public Health policies to community needs. PMID- 15816144 TI - Severe illness and death among injecting drug users in Scotland: a case-control study. AB - Between April and September 2000, 60 injecting drug users in Scotland died or were hospitalized with severe illness. Laboratory investigations suggested that Clostridium novyi and other bacteria were important aetiological agents. To determine associated environmental/behavioural factors a case-control study was undertaken with 19 'definite' and 32 'probable' cases in Glasgow, Scotland. For every deceased case (n=19), up to three proxy individuals were interviewed. Three controls were identified for each case. Multivariate logistic regression analyses compared (i) all cases and controls; (ii) definite cases and matched controls; (iii) probable cases and matched controls. In all three analyses injecting into muscle or skin and injecting most of the time with a filter used by someone else were the variables most strongly associated with illness. Comparing only muscle injecting cases and controls, cases were significantly more likely to have injected larger amounts of heroin per average injection than were controls. The findings make an important epidemiological contribution to the understanding of the public health and clinical implications of the contamination of illicit drugs by histotoxic clostridia. PMID- 15816145 TI - Epidemiology of meningococcal disease in Denmark 1974-1999: contribution of the laboratory surveillance system. AB - The Danish meningococcal disease laboratory surveillance system was established in 1974, based on close collaboration between local Departments of Clinical Microbiology and the Reference Laboratory at Statens Serum Institut. The completeness of the clinical notification system integrated with the laboratory surveillance system has been estimated to be more than 95%. Overall 4257 (79%) of 5356 cases of meningococcal disease notified during 1974-1999 were confirmed by culture of Neisseria meningitidis. The proportion of culture-confirmed cases ranged from 70% in 1989 to 89% in 1980. Only 26 patients (0.6%) with culture confirmed meningococcal disease were not notified. Serological phenotype and susceptibility to penicillin and sulphonamide were determined for all isolates. Multilocus enzyme electrophoresis and/or DNA-based analyses were used for the assessment of clusters and outbreaks. Meningococcal antibody tests and counter immunoelectrophoresis were used for the ascertainment of suspected cases. These combined systems allowed timely and reliable management of outbreaks and identification of clusters. PMID- 15816146 TI - The meningococcal antibody test: how useful in the diagnosis of meningococcal disease? AB - Based on 9257 [correction] blood samples received from 7365 patients with a request for a meningococcal antibody test (MAT) during a 10-year period (1986 1995), the usefulness of the test in the diagnosis of meningococcal disease was assessed. Of 635 patients with culture-confirmed meningococcal disease, 88% were seronegative on admittance to hospital and 90% were seropositive 10-15 days after onset of disease. The humoral immune response in children <2 years of age was weaker than in older children and adults. Among 893 MAT-positive patients without culture-confirmed meningococcal disease, 261 (29%) had been notified as cases of meningococcal disease. Among 228 patients notified as serologically confirmed the MAT results were consistent with the clinical diagnosis in 86%. MAT is a reliable tool for establishing a diagnosis in patients with suspected meningococcal disease. Key factors facilitating appropriate interpretation of negative as well as positive test results were: time(s) of sampling(s) after onset of disease, age of the patient and clinical features. PMID- 15816147 TI - A boarding school outbreak of pertussis in adolescents: value of laboratory diagnostic methods. AB - Culture for Bordetella pertussis (B. pertussis) is the traditional gold standard for laboratory diagnosis of pertussis but is insensitive, especially later in the course of illness and in vaccinated persons. Interpretation of serology is limited by the lack of an appropriate reference standard. An outbreak of pertussis in a crowded boarding-school dormitory allowed evaluation of laboratory correlates of infection. Questionnaires, serum samples and throat swabs were collected from members of the exposed group. Serum samples from unexposed controls of a similar age group were used for comparison. B. pertussis PCR was performed on throat swabs, and sera were tested for IgA antibodies against whole cell (WC) B. pertussis antigen and IgG antibodies to pertussis toxin (PT). The Centers for Disease Control and Prevention definition for pertussis was used to define clinical cases. We evaluated the use of a previously published cut-off for PT IgG of 125 EIA units (EU)/ml. Completed questionnaires were obtained from 115 students, of whom 85 (74%) reported coughing symptoms, including 32 (28%) who met the clinical case definition for pertussis. B. pertussis was detected by PCR in 17 (15%) and WC IgA in 22 (19%) students; neither correlated with symptoms, but dormitory of residence strongly predicted PCR status. The mean PT IgG geometric mean concentration, in this situation of high pertussis exposure, correlated with severity of symptoms and was significantly higher in both symptomatic and asymptomatic children exposed during the outbreak (P < 0.001) than in control children. A cut-off for PT IgG of 125 EU/ml was too high in an outbreak situation to be sensitive enough to identify pertussis cases. A case of pertussis in a crowded boarding-school dormitory resulted rapidly in an outbreak. Serology and PCR were useful in identifying the outbreak and commencing disease control measures. The use of serology has mostly been evaluated in community serosurveys, where it is not possible to determine if immunity reflects vaccination, asymptomatic disease or symptomatic disease. This outbreak gave us the opportunity to evaluate the value of serology and PCR in the presence of confirmed exposure to pertussis. PMID- 15816148 TI - The epidemiology of recurrent bacterial pneumonia in people with AIDS in Europe. AB - Data from AIDS surveillance systems in the World Health Organization European region (1993-2001) were analysed to describe the main epidemiological aspects of recurrent bacterial pneumonia (RBP) as AIDS-defining illness (ADI) in Europe. Among the 153 756 AIDS cases analysed, 5796 (3.8%) had RBP. The proportion of RBP was higher (8.3%) in eastern than in western Europe (3.6%), possibly because of a greater propensity of certain countries to diagnose RBP. In western Europe, the proportion of RBP as ADI appeared to increase over time up to 1998 (from 2.5% to 4.5%), and declined thereafter (3.3% in 2001). RBP was strongly associated with intravenous drug use (odds ratio 3.0, 95% CI 2.7-3.3), whereas it did not differ in age groups or geographical areas. The study findings confirm the crucial role of intravenous drug use in the occurrence of RBP and suggest that highly active antiretroviral therapies mi.ht have had a postponing impact on the relative frequency of RBP as ADI. PMID- 15816149 TI - Incidence of herpes zoster, 1997-2002. AB - We estimated age-specific herpes zoster (HZ) incidence rates in the Kaiser Permanente Northwest Health Plan (KPNW) during 1997-2002 and tested for secular trends and differences between residents of two states with different varicella vaccine coverage rates. The cumulative proportions of 2-year-olds vaccinated increased from 35% in 1997 to 85% in 2002 in Oregon, and from 25% in 1997 to 82% in 2002 in Washington. Age-specific HZ incidence rates in KPNW during 1997-2002 were compared with published rates in the Harvard Community Health Plan (HCHP) during 1990-1992. The overall HZ incidence rate in KPNW during 1997-2002 (369/100,000 person-years) was slightly higher than HCHP's 1990-1992 rate when adjusted for age differences. For children 6-14 years old, KPNW's rates (182 for females, 123 for males) were more than three times HCHP's rates (54 for females, 39 for males). This increase appears to be associated with increased exposure of children to oral corticosteroids. The percentage of KPNW children exposed to oral corticosteroids increased from 2.2% in 1991 to 3.6% in 2002. Oregon residents had slightly higher steroid exposure rates during 1997-2002 than Washington residents. There were significant increases in HZ incidence rates in Oregon and Washington during 1997-2002 among children aged 10-17 years, associated with increased exposure to oral steroids. PMID- 15816150 TI - Influenza and its relationship to circulatory disorders. AB - Three sources of data (general practice episode data from the Weekly Returns Service of the Royal College of General Practitioners, national hospital admission data for England and national mortality data by date of death) were examined separately in each winter (1994/1995 to 1999/2000) to investigate the impact of influenza on circulatory disease. Weekly data on incidence (clinical new episodes) hospital emergency admissions and deaths certified to circulatory disorders and to respiratory diseases (chapters VII and VIII of ICD9) during influenza epidemic periods (defined from combined clinical/virological surveillance) were examined in age groups 45-64, 65-74 and > or =75 years. Data collected in the four winters in which there were substantial influenza A epidemics were consolidated for the period 6 weeks before to 6 weeks after each peak of the epidemic, and associations between the variables at different time lags examined by calculating cross-correlation coefficients. We also examined deaths due to ischaemic heart disease (IHD) as a proportion of all circulatory deaths and deaths due to influenza/pneumonia as a proportion of all respiratory deaths. There were no increases of GP episodes nor of emergency admissions for circulatory disorders in any of the three age groups during epidemic periods. Increased circulatory deaths occurred in all age groups and particularly in the oldest group. The large cross-correlation coefficients of deaths (circulatory and respiratory) with GP respiratory episodes at weekly lags of 0, -1 and 1 were evidence that the deaths and episode distributions were contemporaneous. The ratios of excess circulatory deaths relative to excess respiratory deaths during epidemic periods were 0.74 (age 45-64), 0.72 (65-74) and 0.57 (> or =75 years). Increased circulatory deaths contemporary with new incident cases of respiratory episodes but with no concomitant increase in admissions suggests rapid death during the acute phase of illness. Influenza contingency planning needs to take account of these deaths in determining policy for prophylaxis and in providing facilities for cardio-respiratory resuscitation. PMID- 15816151 TI - Reappearance of influenza B/Victoria/2/87-lineage viruses: epidemic activity, genetic diversity and vaccination efficacy in the Finnish Defence Forces. AB - A new B/Shangdong/7/97-like influenza virus (Victoria/2/87 lineage) predominated during the 2002/2003 epidemic season in Finland and was estimated to account for 2246 of the 13,496 feverish upper respiratory tract infections (URIs) occurring among conscripts in the Finnish army. The incidence (1716/10,000 conscripts) was indicative of moderate epidemic activity at most. Analysis of the cross-reactive antibodies induced in 1988 suggests that the basis of the protection was probably established during the childhood of the conscripts. Vaccination in autumn 2002 prevented 42% of the URIs during the influenza B outbreak and 71% (95% CI 42-85) of infections interpreted as influenza B. Despite the low genetic variability of the Shangdong/7/97-like viruses, breakages of a potential glycosylation site in haemagglutinin (HA1, position 197) were frequent; their biological significance is discussed. The Shangdong/7/97-like strains were HA1/NA reassortants, as were also the less abundant strains that for HA1 belonged to the B/Yamagata/16/88 lineage. A further reassortment, which probably emerged during the outbreak in one of the garrisons, supports our hypothesis that circumstances in these settings may especially favour the emergence of diversity by reassortment. PMID- 15816152 TI - Human coronavirus OC43 causes influenza-like illness in residents and staff of aged-care facilities in Melbourne, Australia. AB - Three outbreaks of respiratory illness associated with human coronavirus HCoV OC43 infection occurred in geographically unrelated aged-care facilities in Melbourne, Australia during August and September 2002. On clinical and epidemiological grounds the outbreaks were first thought to be caused by influenza virus. HCoV-OC43 was detected by RT-PCR in 16 out of 27 (59%) specimens and was the only virus detected at the time of sampling. Common clinical manifestations were cough (74%), rhinorrhoea (59%) and sore throat (53%). Attack rates and symptoms were similar in residents and staff across the facilities. HCoV-OC43 was also detected in surveillance and diagnostic respiratory samples in the same months. These outbreaks establish this virus as a cause of morbidity in aged-care facilities and add to increasing evidence of the significance of coronavirus infections. PMID- 15816153 TI - The transmission dynamics of groups A and B human respiratory syncytial virus (hRSV) in England & Wales and Finland: seasonality and cross-protection. AB - Human respiratory syncytial virus (hRSV) transmission dynamics are inherently cyclical, and the observed genetic diversity (between groups A and B) also appears to have a repeating pattern. A key unknown is the extent to which genetic variants interact immunologically, and thus impact on epidemiology. We developed a novel mathematical model for hRSV transmission including seasonal forcing of incidence and temporary intra- and inter-group partial immunity. Simultaneous model fits to data from two locations (England & Wales, UK, and Turku, Finland) successfully reproduced the contrasting infection dynamics and group A/B dominance patterns. Parameter estimates are consistent with direct estimates. Differences in the magnitude and seasonal variation in contact rate between the two populations alone could account for the variation in dynamics between these populations. The A/B group dominance patterns are explained by reductions in susceptibility to and infectiousness of secondary homologous and heterologous infections. The consequences of the observed dynamic complexity are discussed. PMID- 15816154 TI - An outbreak of viral meningitis associated with a public swimming pond. AB - From July to October 2001, 215 cases of aseptic meningitis occurred among the inhabitants of the German city of Kassel and neighbouring counties. A matched case-control study identified bathing in a public, nature-like pond during the beginning of the outbreak as a risk factor for disease [matched odds ratio (mOR) 44.8, 95% confidence interval (CI) 3.9-515.6]. Among bathers, patients with meningitis spent more time in the water (mOR 18.8, 95% CI 2.0-174.1) and swallowed water more frequently (mOR = 7.3, 95% CI 0.7-81.8). Of 30 cerebrospinal fluid samples tested, echovirus 30 was cultured from 16, and echovirus 13 from seven. An echovirus 30 sequence obtained from one pond water sample showed a 99% nucleotide and 100% amino-acid homology with patient isolates. This outbreak demonstrates the potential of nature-like swimming ponds to cause widespread community infection with substantial public health impact. PMID- 15816155 TI - Seroprevalence of parvovirus B19 infection in daycare educators. AB - This study was undertaken to provide first-time estimates for the seroprevalence of parvovirus B19 infection among daycare educators in Montreal, Canada, and to identify factors associated with seropositivity. A cross-sectional design was used. Directors and educators from 81 daycare centres (DCCs) were surveyed about DCC and personal characteristics respectively, and serum samples from 477 female educators were tested for parvovirus B19 IgG antibodies. The seroprevalence of parvovirus B19 was 70%. Parvovirus B19 seropositivity was significantly associated with age and with working experience in DCCs, but the latter association was restricted to educators aged less than 40 years. In conclusion, working as a daycare educator appears to be associated with increased risk of acquiring parvovirus B19 infection, but this finding will require further investigation. Because of the large proportion of educators susceptible to acquiring parvovirus B19 infection, our findings also highlight the need for preventive measures. PMID- 15816156 TI - Management of chronic hepatitis C in French departments of internal medicine and infectious diseases. AB - This prospective, multicentre study was conducted during 2-30 April 2001 in the internal medicine/infectious diseases services in France and included data from 1858 hepatitis C virus (HCV)-infected patients, half of whom were HIV co infected. The aims were to outline the type of pre-therapeutic evaluation of HCV infection performed (HCV RNA, genotype, liver biopsy); determine the proportion and characteristics of patients receiving antiviral treatment; and determine if any changes in these parameters had occurred between 1995 and 2001. Patients whom had a complete pre-therapeutic evaluation (39%, 709/1834) and received antiviral treatment (38%, 690/1830) were more likely to have abnormal liver biochemistry, cirrhosis and cryoglobulinaemia (P < 0.001). Injecting drug users and HIV-co infected patients were less likely to have a complete pre-therapeutic evaluation or receive antiviral treatment (P < 0.001). A complete pre-therapeutic evaluation was more often performed in 2001 than in 1995 (39% vs. 6%, P < 0.001), including qualitative HCV RNA testing (91% vs. 68%, P < 0.001), genotyping (59% vs. 7%, P < 0.001) and a liver biopsy (60% vs. 29%, P < 0.001). The frequency of anti-HCV treatment approximately doubled between 1995 and 2001 (20% vs. 38%, P < 0001). Although adherence to consensus recommendations regarding pre-therapeutic evaluation is not ideal, a substantial improvement has occurred since 1995. Nevertheless, means of increasing the availability of antiviral therapies, particularly for patients with HIV co-infection or injecting drug use, require further study. PMID- 15816157 TI - Seroepidemiology of Trichomonas vaginalis in rural women in Zimbabwe and patterns of association with HIV infection. AB - Serological assays using dried blood spots from 5221 women in rural areas of eastern Zimbabwe were used to assess the epidemiology of Trichomonas vaginalis infection, and its association with HIV. Antibodies to T. vaginalis and to HIV were detected by enzyme immunoassays. Behavioural and demographic data were collected by confidential questionnaires. In total, 516 (9.9%) women were seropositive for T. vaginalis and seroprevalence increased with age among younger women. Divorced, widowed and single women were more likely to be seropositive. After controlling for age, seropositivity was significantly associated with being sexually active, having multiple sex partners, having a partner who had multiple sex partners, and having a new sex partner in the past year. Seropositivity was associated with a recent history of genital discharge. Overall, 208 (40.3%) T. vaginalis-positive samples were also positive for HIV, compared with 1106 (23.5%) T. vaginalis-negative samples (age and sex adjusted OR 2.11, 95% CI 1.74-2.55, P < 0.001). There was increased risk for being HIV-positive amongst T. vaginalis seropositive women regardless of residence, employment or education. In a logistic regression controlling for common risk factors, the association remained significant. T. vaginalis-seropositive young women with a history of genital discharge were much more likely to be HIV-positive than women who were T. vaginalis-seronegative and had no history of discharge (OR 6.08, 95 % CI 2.95 12.53). Although a causal relationship cannot be assumed, detection and treatment of trichomoniasis may be important in strategies to reduce HIV transmission through sexually transmitted infection control. PMID- 15816158 TI - Distribution and susceptibility to amphotericin B and fluconazole of Candida spp. isolated from Taiwan. AB - Susceptibilities to amphotericin B and fluconazole of 628 clinical yeast strains collected from 22 hospitals in Taiwan were determined. A total of 53 isolates (8.4%) were resistant to fluconazole. Each hospital had different resistance rate to fluconazole ranging from 0% to 24%. None of the 186 isolates from eight of the 22 hospitals was resistant to fluconazole. In contrast, isolates from nine of the remaining 14 hospitals had greater than 10% resistance rate to fluconazole. Consistently, 88.9% (8/9) fluconazole-resistant C. albicans isolates were from hospitals having a high resistance rate to fluconazole. The prevalence of various Candida spp. in each hospital was different. A positive association was found between the prevalence of C. tropicalis and the resistance rate to fluconazole for individual hospitals. Although only three isolates (0.5%) were resistant to amphotericin B, a co-resistance to both amphotericin B and fluconazole was observed, which highlights the emerging problem of drug resistance. PMID- 15816159 TI - Seroepidemiology of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in wild mice captured in northern Turkey. AB - An expedition across the Asian part of the Black Sea coast and national parks of Northern Turkey was organized in the summer of 2001 to investigate the presence of Borrelia burgdorferi sensu lato (s.l.), Lyme borreliosis agent, and Anaplasma phagocytophilum, human granulocytic ehrlichiosis, agent, in wild mice. A total of 65 Apodemus flavicollis, Apodemus sylvaticus, Microtus epiroticus, Crocidura suaveolens and Mus macedonicus, were captured. Two out of 22 Apodemus sylvaticus specimens were seropositive for B. afzelii by enzyme-linked immunosorbent assay as confirmed by Western blotting, however cultures of skin and bladder samples from all small mammals in Barbour-Stoenner-Kelly's medium-II remained negative for B. burgdorferi s.l. All sera tested were negative for Anaplasma phagocytophilum by indirect immunofluorescent assay. The prevalence of B. burgdorferi s.l. and Anaplasma phagocytophilum is low in wild mice of the Asian part of Northern Turkey. PMID- 15816160 TI - Modelling and determination of the transmission contact rate for contagious bovine pleuropneumonia. AB - Contagious bovine pleuropneumonia (CBPP) is a cattle respiratory disease that represents one of the major threats to cattle health and production in sub Saharan Africa. The transmission contact rate of CBPP plays a key role in the spreading dynamics of the disease. We have developed an approach based on the combination of a SEIR model describing the spread of CBPP with the dynamic of seroconversion to determine the transmission contact rate for CBPP. This method has been subsequently applied to serological diagnostic data obtained from an experimental vaccine trial. As a result, we find that the transmission contact rates for subclinical, clinical and chronic infective states are respectively, 0.084/N, 0.45 and 0.14/N per animal per day, where N is the herd population size, and the basic reproductive number corresponding to this trial (N=28) is R0=27. PMID- 15816161 TI - Clustering of equine grass sickness cases in the United Kingdom: a study considering the effect of position-dependent reporting on the space-time K function. AB - Equine grass sickness (EGS) is a largely fatal, pasture-associated dysautonomia. Although the aetiology of this disease is unknown, there is increasing evidence that Clostridium botulinum type C plays an important role in this condition. The disease is widespread in the United Kingdom, with the highest incidence believed to occur in Scotland. EGS also shows strong seasonal variation (most cases are reported between April and July). Data from histologically confirmed cases of EGS from England and Wales in 1999 and 2000 were collected from UK veterinary diagnostic centres. The data did not represent a complete census of cases, and the proportion of all cases reported to the centres would have varied in space and, independently, in time. We consider the variable reporting of this condition and the appropriateness of the space-time K-function when exploring the spatial temporal properties of a 'thinned' point process. We conclude that such position dependent under-reporting of EGS does not invalidate the Monte Carlo test for space-time interaction, and find strong evidence for space time clustering of EGS cases (P < 0.001). This may be attributed to contagious or other spatially and temporally localized processes such as local climate and/or pasture management practices. PMID- 15816162 TI - Prevalence of verotoxin-producing Escherichia coli (VTEC) 0157 in Swedish dairy herds. AB - A prevalence study of verotoxin-producing Escherichia coli O157 (VTEC O157) was performed in 371 randomly selected dairy herds distributed throughout Sweden. Faecal and manure samples were collected and analysed by immunomagnetic separation and culturing. Data were recorded for each herd regarding herd size, age of sampled animals and whether, in addition to cattle, the farm kept other animals. VTEC O157 was isolated from 33 (8.9%) of the 371 investigated herds. The prevalence was higher (23.3%) in Halland county than in the rest of Sweden (P > 0.01). Halland was also the county in Sweden that during the study period had the highest incidence of human VTEC O157 cases. VTEC O157 was not detected on any farm in northern Sweden. Identified risk factors, in the multivariate analyses, for herds being VTEC O157 positive were herd size, geographical localization, presence of pigs on the farm and median age of sampled animals. PMID- 15816163 TI - The prevalence of Shiga toxin-producing Escherichia coli in domestic animals and food in Serbia. AB - Faecal samples of 2660 domestic animals from 116 farms and 956 samples of food were examined for the presence of Shiga toxin-producing Escherichia coli (STEC). STEC was recovered from 126 (15.3%) cattle, 135 (11.3%) pigs, 135 (66.8%) sheep, 31 (73.8%) goats, 4 (1%) chicken, and 15 (1.6%) food samples. Of all STEC isolates, 21.5, 25.8 and 15% produced enterohaemolysin, alpha-haemolysin, and aerobactin respectively, 1.6% displayed localized adherence (LA) to HEp-2 cells, 27.6% were sorbitol negative, and 30% were resistant to antibiotics. Only 14 (3.1%) of the STEC isolates belonged to human infection-associated serogroups (O26, O55, O111, O128 and 0157), designated as enterohaemorrhagic E. coil (EHEC). This study revealed that STEC are prevalent in domestic animals, and to a lesser extent in food of animal origin in Serbia, but the absence of a EHEC phenotypic profile (characteristic serogroup, LA, enterohaemolysin production) in most animal STEC strains may explain the low incidence of human STEC infection in this part of the world. PMID- 15816164 TI - High-level aminoglycoside resistant enterococci isolated from swine. AB - Approximately 42% (187/444) of swine enterococci collected between the years 1999 and 2000 exhibited high-level resistance to gentamicin (MIC > or =500 microg/ml), kanamycin (MIC > or =500 microg/ml), or streptomycin (MIC > or =1000 microg/ml). Eight aminoglycoside resistance genes were detected using PCR, most frequently ant(6)-Ia and aac(6')-Ii from Enterococcus faecium. Twenty-four per cent (45/187) of total high-level aminoglycoside-resistant isolates and 26% (4/15) of isolates resistant to high levels of all three antimicrobials were negative for all genes tested. These data suggest that enterococci isolated from swine contain diverse and possibly unidentified aminoglycoside resistance genes. PMID- 15816165 TI - An outbreak of multidrug-resistant Salmonella enterica serotype Newport infections linked to the consumption of imported horse meat in France. AB - In 2003, 14 cases of multidrug-resistant Salmonella Newport infections were reported. This is the first documented foodborne outbreak of multidrug-resistant S. Newport in France. The blaCMY gene was present in all isolates. All cases reported having eaten horse meat from a common wholesaler. The country of origin of the imported meat could not be identified. PMID- 15816166 TI - An outbreak of Q fever in a prison in Italy. AB - We observed an outbreak of Q fever in a prison population. Overall, 65 of the 600 prison inmates developed the disease. The location of the prison cells had no apparent effect on the risk of infection. The outbreak was probably due to exposure to dust contaminated by a passing flock of sheep, which at the time of the outbreak was engaged in lambing. These findings highlight the possible emergence of Q fever in settings and populations not normally thought of as being at risk of exposure to the infection. PMID- 15816167 TI - Fifty years of progress in ion channel research. AB - Fifty years ago, ion channels were but a reasonable hypothesis. I outline some major steps in transforming this idea from a plausible description of the biological assemblies responsible for controlling passive ion transport across membranes to established fact. Important electrophysiological, biochemical, molecular biological, structural, and theoretical tools are discussed in the context of the transition from studying whole cell preparations, containing many channels, to investigating single channel behavior. Six channel families are exemplified: the model peptide, gramicidin, the acetylcholine receptor, and the sodium, potassium, calcium, and chloride channels. Some questions of current interest are posed. PMID- 15816168 TI - Gramicidin channels. AB - Gramicidin channels are mini-proteins composed of two tryptophan-rich subunits. The conducting channels are formed by the transbilayer dimerization of nonconducting subunits, which are tied to the bilayer/solution interface through hydrogen bonds between the indole NH groups and the phospholipid backbone and water. The channel structure is known at atomic resolution and the channel's permeability characteristics are particularly well defined: gramicidin channels are selective for monovalent cations, with no measurable permeability to anions or polyvalent cations; ions and water move through a pore whose wall is formed by the peptide backbone; and the single-channel conductance and cation selectivity vary when the amino acid sequence is varied, even though the permeating ions make no contact with the amino acid side chains. Given the amount of experimental information that is available--for both the wild-type channels and for channels formed by amino acid-substituted gramicidin analogues--gramicidin channels provide important insights into the microphysics of ion permeation through bilayer-spanning channels. For the same reason, gramicidin channels constitute the system of choice for evaluating computational strategies for obtaining mechanistic insights into ion permeation through the complex channels formed by integral membrane proteins. PMID- 15816169 TI - Potassium channels. AB - Potassium channels are integral membrane proteins that selectively transport K+ across the cell membrane. They are present in all mammalian cells and have a wide variety of roles in both excitable and nonexcitable cells. The phenotypic diversity required to accomplish their various roles is created by differences in conductance, the timecourse and mechanisms of different gating events, and the interaction of channels with a variety of accessory proteins. Through the integration of biophysical, molecular, structural, and theoretical studies, significant progress has been made toward understanding the structural basis of K+ channel function, and diseases associated with K+ channel dysfunction. PMID- 15816170 TI - Voltage-gated ion channels. AB - Voltage-dependent ion channels are membrane proteins that conduct ions at high rates regulated by the voltage across the membrane. They play a fundamental role in the generation and propagation of the nerve impulse and in cell homeostasis. The voltage sensor is a region of the protein bearing charged amino acids that relocate upon changes in the membrane electric field. The movement of the sensor initiates a conformational change in the gate of the conducting pathway thus controlling the flow of ions. Major advances in molecular biology, spectroscopy, and structural techniques are delineating the main features and possible structural changes that account for the function of voltage-dependent channels. PMID- 15816171 TI - Unique structure and function of chloride transporting CLC proteins. AB - CLC proteins are a large structurally defined family of Cl- ion channels and H+/Cl- antiporters with nine distinct genes in mammals. The membrane-embedded part of CLC proteins bears no obvious similarity to any other class of membrane proteins, while the cytoplasmic C-terminus of most eukaryotic and some prokaryotic CLCs contains two regions with homology to cystathionine beta synthase (CBS) domains that are found in other proteins as well. Different members serve a broad range of physiological roles, including stabilization of the membrane potential, transepithelial ion transport, and vesicular acidification. Their physiological importance is underscored by the causative involvement in at least four different human genetic diseases. From functional studies of the Torpedo homologue ClC-0, a homodimeric architecture with two physically separate ion conduction pathways was anticipated and fully confirmed by solving the crystal structure of prokaryotic CLC homologues. The structure revealed a complex fold of 18 alpha-helices per subunit with at least two Cl- ions bound in the center of each protopore. A critical glutamic acid residue was identified whose side-chain seems to occupy a third Cl- ion binding site in the closed state and that moves away to allow Cl- binding. While the overall architecture and pore structure is certainly conserved from bacteria to humans, the bacterial proteins that were crystallized are actually not Cl- ion channels, but coupled H+/Cl- antiporters. These recent breakthroughs will allow us to study in further detail the structure, function, and the physiological and pathophysiological role of CLC proteins. PMID- 15816172 TI - Voltage-gated sodium and calcium channels in nerve, muscle, and heart. AB - Voltage-gated ion channels are membrane proteins which underlie rapid electrical signals among neurons and the spread of excitation in skeletal muscle and heart. We outline some recent advances in the study of voltage-sensitive sodium and calcium channels. Investigations are providing insight into the changes in molecular conformation associated with open-closed gating of the channels, the mechanisms by which they allow only specific ion species to pass through and carry an electric current, and the pathological consequences of small perturbations in channel structure which result from genetic mutations. Determination of three-dimensional structures, coupled with molecular manipulations by site-directed mutagenesis, and parallel electrophysiological analyses of currents through the ion channels, are providing an understanding of the roles and function of these channels at an unprecedented level of molecular detail. Crucial to these advances are studies of bacterial homologues of ion channels from man and other eukaryotes, and the use of naturally occurring peptide toxins which target different ion channel types with exquisite specificity. PMID- 15816173 TI - Ligand-gated channels. AB - Ligand-gated ion channels (LGICs) are fast-responding channels in which the receptor, which binds the activating molecule (the ligand), and the ion channel are part of the same nanomolecular protein complex. This paper will describe the properties and functions of the nicotinic acetylcholine LGIC superfamily, which plays a critical role in the fast chemical transmission of electrical signals between nerve cells and between nerve and muscle cells. The superfamily will mainly be exemplified by the excitatory nicotinic acetylcholine receptor (nAChR) and the inhibitory glycine receptor (GlyR) channels. PMID- 15816174 TI - Poisson-Nernst-Planck theory approach to the calculation of current through biological ion channels. AB - The Poisson-Nernst-Planck (PNP) theory of electro-diffusion is reviewed. Techniques for numerical solution of the three-dimensional PNP equations are summarized, and several illustrative applications to ion transport through protein channels are presented. Strengths and weaknesses of the theory are discussed, as well as attempts to improve it via increasingly realistic evaluation of the force acting on each ion due to the protein/membrane environment. PMID- 15816175 TI - Semimicroscopic modeling of permeation energetics in ion channels. AB - The semimicroscopic (SMC) approach to modeling the energetics of ion permeation through biological channels provides an alternative perspective to standard molecular dynamics methods. It exploits the timescale separation between electronic and structural contributions to dielectric stabilization and accounts for electronic polarization by embedding the channel in a milieu that, on average, describes this polarization. Ions, water, and selected peptide moieties are mobile and comprise the reorganizational contribution to dielectric stabilization. The conceptual advantages and limitations of the technique are described. Methodological details are outlined, stressing three convenient electrical geometries. Practical aspects of the SMC procedure are explained, highlighting the areas ripe for further development. Finally, some specific applications are considered. PMID- 15816176 TI - Brownian dynamics simulation for modeling ion permeation across bionanotubes. AB - The principles underlying Brownian dynamics (BD), its statistical consistency, and algorithms for practical implementation are outlined here. The ability to compute current flow across ion channels confers a distinct advantage to BD simulations compared to other simulation techniques. Thus, two obvious applications of BD ion channels are in calculation of the current-voltage and current-concentration curves, which can be directly compared to the physiological measurements to assess the reliability of the model and predictive power of the method. We illustrate how BD simulations are used to unravel the permeation dynamics in two biological ion channels-the KcsA K+ channel and CIC Cl- channel. PMID- 15816177 TI - Molecular dynamics simulation approaches to K channels: conformational flexibility and physiological function. AB - Molecular modeling and simulations enable extrapolation for the structure of bacterial potassium channels to the function of their mammalian homologues. Molecular dynamics simulations have revealed the concerted single-file motion of potassium ions and water molecules through the selectivity filter of K channels and the role of filter flexibility in ion permeation and in "fast gating." Principal components analysis of extended K channel simulations suggests that hinge-bending of pore-lining M2 (or S6) helices plays a key role in K channel gating. Based on these and other simulations, a molecular model for gating of inward rectifier K channel gating is presented. PMID- 15816179 TI - New direction for health care regulation? PMID- 15816178 TI - Microchip technology in ion-channel research. AB - The electrical activity of living cells can be monitored in various ways, but for the study of ion channels and the drugs that affect them, the patch-clamp techniques are the most sensitive. Recent developments in microfabricated patch clamp electrodes are reviewed, and technical challenges for the future are discussed. PMID- 15816180 TI - Efficacy of a recombinant equine influenza vaccine against challenge with an American lineage H3N8 influenza virus responsible for the 2003 outbreak in the United Kingdom. AB - Fifteen influenza-naive Welsh mountain ponies were randomly assigned to three groups of five. A single dose of a recombinant ALVAC vaccine was administered intramuscularly to five of the ponies, two doses, administered five weeks apart, were administered to five, and the other five served as unvaccinated, challenge controls. Two weeks after the completion of the vaccination programme, the ponies were all challenged by exposure to an aerosol of influenza virus A/eq/Newmarket/5/03. Their clinical signs were scored daily for 14 days according to a standardised scoring protocol, and nasal swabs were taken daily for 10 days to monitor the excretion of virus. The challenge produced severe clinical signs of influenza (fever, coughing, nasal discharge and dyspnoea) in all five control ponies, but the vaccinated ponies developed only mild disease, consisting of a serous nasal discharge lasting for only one day. The excretion of virus was almost completely suppressed in the vaccinated ponies, but the control ponies shed the virus for up to seven days after the challenge. PMID- 15816181 TI - Effect of vaginal and cervical deposition of semen on the fertility of sheep inseminated with frozen-thawed semen. AB - The effect of vaginal and cervical deposition of frozen-thawed semen on the fertility of sheep was tested in a field trial in which 543 Norwegian crossbred ewes aged between six months and five-and-a-half years from 10 farms were inseminated after natural oestrus. Cervical insemination with 200 x 10(6) spermatozoa resulted in 25-day non-return and lambing rates of 75.4 and 72.7 per cent, respectively, and vaginal insemination gave rates of 71.3 and 67.4 per cent; the cervical inseminations produced significantly higher lambing rates (P=0.04). There were significant differences between the lambing rates for different rams (P=0.006) and different farmers (P=0.003), and there was a significant interaction between farmer and deposition site (P=0.03). After vaginal insemination fertility was encouragingly high, but the results varied with the farmer, and different flock and management conditions. PMID- 15816182 TI - Serological evidence of Leptospira interrogans serovar Bratislava infection and its association with abortions in cattle in northern Spain. AB - A cross-sectional study was carried out in the Basque Country of Spain to determine the seroprevalence of 10 Leptospira serovars in a population of dairy cattle with poor fertility, and a case-control study was carried out in another northern area to investigate the role of Leptospira interrogans serovar Bratislava in abortions. L. Bratislava was the most prevalent serovar in the cross-sectional study, with 25.4 per cent of the cows testing positive in the microagglutination test when a cut-off of 1:10 or higher was applied, followed by Leptospira Hardjo (8.2 per cent), Leptospira Pomona (7.7 per cent), Leptospira Autumnalis (0.7 per cent) and Leptospira Copenhageni (0.1 per cent). In the case control study the seroprevalence of L. Bratislava was significantly higher among the cows which had aborted when a titre of 1:300 or more was used as a cut-off (9.7 per cent v 3.4 per cent, P=0.008); 69 per cent of the L. Bratislava-infected cows that had aborted apparently aborted as a result of the infection. PMID- 15816183 TI - 'Typical' veterinary consultation in the United Kingdom and Ireland. PMID- 15816184 TI - Isolation of verocytotoxin-producing Escherichia coli O157 from water buffaloes (Bubalus bubalis) in southern Italy. PMID- 15816185 TI - Lack of evidence of porcine circovirus type 1 and type 2 infection in piglets with congenital tremors in Korea. PMID- 15816186 TI - Intra-auricular trichomonosis in a saker falcon (Falco cherrug) in Saudi Arabia. PMID- 15816187 TI - Use of lectin histochemistry to diagnose Sida carpinifolia (Malvaceae) poisoning in sheep. PMID- 15816188 TI - Granulomatous lesion on a bitch's nipple caused by Leishmania infantum. PMID- 15816189 TI - Market forces and animal welfare. PMID- 15816190 TI - Prescribing of proposed POM-VPM products. PMID- 15816191 TI - Mycoplasma bovis isolated from brain tissue of calves. PMID- 15816192 TI - Acute patella fractures in cats. PMID- 15816193 TI - Pathogenesis of experimental bovine spongiform encephalopathy: preclinical infectivity in tonsil and observations on the distribution of lingual tonsil in slaughtered cattle. AB - The infectivity in tissues from cattle exposed orally to the agent of BSE was assayed by the intracerebral inoculation of cattle. In addition to the infectivity in the central nervous system and distal ileum at stages of pathogenesis previously indicated by mouse bioassay, traces of infectivity were found in the palatine tonsil of cattle killed 10 months after exposure. Because the infectivity may therefore be present throughout the tonsils in cattle infected with BSE, observations were made of the anatomical and histological distribution of lingual tonsil in the root of the tongue of cattle. Examinations of tongues derived from abattoirs in Britain and intended for human consumption showed that macroscopically identifiable tonsillar tissue was present in more than 75 per cent of them, and even in the tongues in which no visible tonsillar tissue remained, histological examination revealed lymphoid tissue in more than 90 per cent. Variations in the distribution of the lingual tonsil suggested that even after the most rigorous trimming of the root of the tongue, traces of tonsillar tissue may remain. PMID- 15816194 TI - Serosurvey of Aujeszky's disease virus infection in European wild boar in Spain. AB - Serum samples from 693 hunted wild boar (Sus scrofa) were analysed by means of a blocking ELISA technique, and the mean (se) prevalence of antibodies to Aujeszky's disease virus was 44 (4) per cent. All the seropositive wild boar were from south central Spain, except for one from central Spain, close to the main positive area. In this area, where large game species are increasingly managed for hunting, the seroprevalence was affected by the type of management. More intensively managed populations had a higher prevalence than wild boar living in natural situations, and the seroprevalence increased with the age of the animals; the seroprevalence was higher in females in all age groups. The seroprevalence in males more than one year old peaked after the breeding season, whereas females of the same age had a higher and constant seroprevalence throughout the year. PMID- 15816195 TI - Investigations of D-lactate metabolism and the clinical signs of D-lactataemia in calves. AB - Five clinically healthy calves received an intravenous injection of 25 g sodium D lactate (223 mmol) in 100 ml sterile water and five control calves were given the same volume of 0.9 per cent sodium chloride. Two clinical examiners who were blinded to the status (test or control) of the calves observed that between eight and 40 minutes after the injections the calves that had received sodium-D-lactate could be distinguished with certainty from the control calves on the basis of their clinical signs, for example, an impaired palpebral reflex, somnolence and a staggering gait. One-compartment and two-compartment analyses of the changes in the plasma concentration of D-lactate, and its renal clearance, indicated that the calves metabolised considerable amounts of D-lactate. PMID- 15816196 TI - Expansion of Culicoides imicola, the main bluetongue virus vector in Europe, into Catalonia, Spain. PMID- 15816197 TI - Prevalence of sarcocystis species in slaughtered goats in Shiraz, Iran. PMID- 15816198 TI - Louping ill in llamas (Lama glama) in the Hebrides. PMID- 15816199 TI - Detection of Cryptosporidium andersoni in a dairy cow with diarrhoea in the UK. PMID- 15816200 TI - Bovine TB and badgers. PMID- 15816201 TI - 24-hour emergency cover. PMID- 15816202 TI - Isolation and identification of goose parvovirus in the UK. PMID- 15816203 TI - Infections with Corynebacterium diphtheriae: six years' experience at an inner London teaching hospital. AB - Recently, a preponderance of non-toxigenic strains of Corynebacterium diphtheriae has been reported, as has the broadening spectrum of disease caused by these strains. This study presents data on 85 isolates of C. diphtheriae over a six year period (1998-2003). Eighty were non-toxigenic isolates from patients with sore throat, and five (one toxigenic) were from cutaneous ulcers in travellers returning from endemic areas. When examined in relation to denominator data provided by the Health Protection Agency (HPA) for the whole of England and Wales, 1998-2002, 75% of all notifications of C. diphtheriae for England and Wales originated from the laboratories at University College London Hospitals (UCLH). In some years (1999 and 2001) 95-100% of isolates came from UCLH. We believe that national data do not reflect true incidence, as universal screening for these organisms is not routine policy in many laboratories. The results presented suggest the need for increased clinical and laboratory awareness of this important pathogen. PMID- 15816204 TI - Progressive decrease of proinsulin secretion in sulphonylurea-treated type 2 diabetes. AB - Progressive deterioration of beta-cell function is proposed as a disease-related factor of sulphonylurea (SU) failure in type 2 diabetes. If it gradually worsens over time then disease duration may mirror the progressive beta-cell deterioration. The aim of the present study is to assess whether or not disease duration is influential in remodelling the secretion pattern of insulin-like molecules and in glucose control of SU-treated type 2 diabetes. A research model is used to investigate proinsulin secreting capacity over time, using two groups of patients: i) disease duration <5 years (n=62), comprising SU responders (SUr; n=48) and SU failures (SUf; n=14); and ii) disease duration > or = 5 years (n= 37), comprising an SUr group (n=17) and an SUf group (n=20). Blood samples are taken at 0 h, 0.5 h 1 h, 2 h and 3 h during a standard oral glucose tolerance test and measured for glucose, total proinsulin (TPI), intact proinsulin (IPI) and specific insulin (SI) concentrations. Pairwise comparison of estimated marginal means of blood glucose, SI, IPI and TPI levels at each time point are carried out between groups and subgroups. (SUr vs. SUf). Homa insulin resistance index (IR index) is applied to analyse IR between the groups. It was found that patients with shorter disease duration had higher proinsulin (TPI and IPI) levels at all time points (P<0.05), together with a lower glucose level at 2 h and 3 h (P<0.05). Homa insulin index analysis showed no difference between the two groups (P=0.26). Results also showed that the SUr group had a significantly lower glucose level at Oh and 3h (P<0.05), although no significant difference in insulin and proinsulin levels was found between the SUr and SUf groups. In conclusion, proinsulin may play an important role in glucose control in SU treated type 2 diabetes, but the effect is reduced in SUf patients. PMID- 15816205 TI - Evaluation of a distance-learning immunology and pathology module in a postgraduate biomedical science course. AB - An electronic presentation of materials for a distance-learning immunology and pathology module from a postgraduate biomedical science course is evaluated. Two different electronic presentation formats for the delivery of the educational material to distance learners are assessed. Responses from users of this material highlighted a preference for a format that has a design tailored to distance learning. There was no significant difference in learning outcome between those taking the module on campus and by distance learning. This suggests that the prerequisites for entry, learning materials and direction given to the students studying by distance learning are adequate for these students to achieve the learning objectives outlined in the course. The evaluation also gave direction for areas within the (CAL) application that can be improved for future students. PMID- 15816206 TI - Low frequency of elevated prothrombin times in patients with lupus anticoagulants when using a recombinant thromboplastin reagent: implications for dosing and monitoring of oral anticoagulant therapy. AB - Many patients with lupus anticoagulants (LA) are treated with oral anticoagulation and monitored using the international normalised ratio (INR) derived from the prothrombin time (PT). Recent reports have produced conflicting conclusions about the extent to which LA interferes with PT determination. The degree of anticoagulation may be overestimated in a patient whose LA affects the PT. A number of reports conclude that specific thromboplastin reagents containing recombinant tissue factor are sensitive to the presence of LAs and should not be used to monitor oral anticoagulant therapy in these patients. These studies were performed on orally anticoagulated patients. The present retrospective study on 400 patients with LAs who were not receiving therapeutic anticoagulation was performed to ascertain the frequency of prolonged PT in these patients when using Innovin recombinant thromboplastin. Only 17 (4.3%) out of 400 had prolonged PT in the presence of LA. As this is a low prevalence, and not all patients with LAs will require anticoagulant therapy, it is concluded that baseline INR determination should be used to highlight the need to monitor individual patients with LA-insensitive reagents. As the use of moderate-intensity oral anticoagulation for patients with LAs and previous thrombosis is receiving wider acceptance, an informed approach to anticoagulant monitoring will reduce the possibility of under-anticoagulating patients receiving this therapy. PMID- 15816207 TI - Antigenaemia and antibody response to Toxoplasma gondii in human immunodeficiency virus-infected patients. AB - Toxoplasma encephalitis in immunocompromised patients results from reactivation of previously acquired (latent) infection. The aim of the study is to assess the antigenaemia and antibody response to Toxoplasma gondii in human immunodeficiency virus (HIV)-infected patients to determine the best marker for early diagnosis of toxoplasmosis in such patients. Indirect enzyme-linked immunosorbent assay (ELISA) for detection of IgG, IgM and IgA anti-toxoplasma antibodies and double sandwich ELISA for toxoplasma antigen is carried out in serum samples collected from 100 HIV seropositive patients and 75 controls. Toxoplasma-specific IgG, IgM and IgA antibody response and antigenaemia were detected in 12%, 6%, 7% and 14% of HIV-infected patients, respectively. On retrospective analysis of 14 patients with antigenaemia only one had central nervous system (CNS) symptoms attributable to toxoplasma infection. In this patient, the CD4+ cell count was below 50/microL and none of the specific immunoglobulin isotype responses could be detected. The patient showed clinical improvement following specific chemotherapy for toxoplasmosis. In 25 HIV-negative and anti-toxoplasma IgG antibody-positive controls, IgM was detected in two (8%), IgA in five (20%) and antigenaemia in 10 (40%), while 50 HIV seronegative healthy controls were negative for both antigen and antibody responses. The study indicates that detection of toxoplasma antigen in addition to IgG antibody response may prove to be a useful indicator in the early diagnosis of reactivated toxoplasmosis in HIV/AIDS patients. PMID- 15816208 TI - Frequency and distribution of group I intron genotypes of Candida albicans colonising critically ill patients. AB - A study is performed to examine the distribution and frequency of 25S rRNA intron genotypes of Candida albicans isolated from different anatomical sites of patients in an intensive care unit (ICU) setting. Germ-tube positive Candida isolates (n=65) from 65 patients are included and isolates are characterised by 25S intron genotyping, whereby all can be subdivided into four genotypes (A-D). Results demonstrated that there were no significant differences between the frequency and genotype distribution of the Candida isolates and the anatomical site of colonisation. Furthermore, analysis of the transposable intron region in the 25S rRNA gene demonstrated equal distribution, regardless of age and anatomical site of isolation (groin, throat, etc.). Therefore, there does not appear to be any selective pressure associated with any anatomical site, resulting in an ecological shift in the frequency of genotypes present. This suggests that C. albicans intron genotypes equally colonise those sites of the body examined in this study. Although such an ecological finding as this is interesting, it perpetuates the continued need to find a genotypic typing scheme that helps to identify the source (nosocomial or endogenous) and mode of entry of C. albicans into patients in the ICU setting, resulting in C. albicans bloodstream infection. PMID- 15816209 TI - Erythromycin resistance among invasive pneumococci in Scotland, 1994-2003. PMID- 15816210 TI - Comparison of in vitro susceptibilities to levofloxacin and ciprofloxacin with Pseudomonas aeruginosa and Stenotrophomonas maltophilia isolated from cystic fibrosis patients in Northern Ireland. PMID- 15816211 TI - Acquired immune deficiency syndrome and human immunodeficiency virus: Taiwanese medical laboratory technologists and students' attitudes, concerns and knowledge. PMID- 15816212 TI - Sequencing and analysis of the 16S rDNA of thermophilic Campylobacter lari and their reliability for molecular discrimination. PMID- 15816213 TI - Alkhumra haemorrhagic fever: case report and infection control details. PMID- 15816214 TI - Bacteria as potential tools in bioterrorism, with an emphasis on bacterial toxins. AB - The threat of bioterrorism remains a reality worldwide and, although of low probability, an attack would be a high-consequence event. Microbes are available to individuals with appropriate contacts and even many low-grade bacterial pathogens can severely affect health. Toxins provide bacteria with a system of defence that is often detrimental to humans and their versatility makes them potential tools of bioterrorism. It should be remembered that the aim of terrorism is not always to kill but rather to strike fear into peoples lives. Therefore, agents such as botulinum and cholera toxin could be used, which may not cause significant mortality but would cause widespread panic and potentially high morbidity. Importantly, no state can ever be fully prepared for a response and it is probable that no state ever could be. It is for this reason that biological agents are so attractive as weapons. PMID- 15816215 TI - [Silence is the sun that ripens the soul's fruits]. PMID- 15816216 TI - [The questions. The difficult academic and professional balance in "pre-European" Spain. (I)]. AB - The supposition that the body of knowledge transmitted by academics adequately prepares students for the demands of daily practice must be revised. The technical nature of academic knowledge and its supposed scientific basis provide an abstract and normative body of knowledge which is only useful as a means to solve patients' problems in a few very concrete moments in professional practice since this profession is unstable, uncertain, complex and saturated with value options. PMID- 15816217 TI - [Hydro-colloidal dressings which release hydro-active silver]. AB - The study presented is a multicentric, prospective, open and comparative study designed with the objective of evaluating the performance of an antibacterial hydrocolloid dressing with hydroactivated silver (Comfeel Plata), when used to activate the healing process in wounds with high bacterial load, clinical signs of infection or malodour. Additionally, once the wound bed was appropriately prepared, a comparison in terms of efficacy was made between, on the one hand, continued treatment with the antibacterial hydrocolloid dressing, and, on the other hand, continued treatment with other dressings specifically designed for the proliferative phase of healing. Included into this study were 43 patients with chronic ulcers who were divided into two parallel treatment groups: In one group, Comfeel Plata (Coloplast AIS) was used until complete wound healing or for a maximum of 10-12 weeks, and in the second group Comfeel Plata (Coloplast A/S) was used until a clean wound bed was obtained and until the wound showed signs of positive evolution, at which moment the treatment was continued until complete healing or for a maximum of 10-12 weeks with dressings without silver designed especially for the proliferative phase of healing [Alione, Comfeel or Biatain (Coloplast AIS)]. The results obtained from the various study parameters indicate that the use of Comfeel Plata in the treatment of infected or colonized wounds prepares the wound bed and facilitates more rapid healing, and that the use of Comfeel Plata effectively reduces pain and malodour. The results indicate that once a clean wound bed is obtained, the use of a dressing without silver specifically for the proliferative phase will facilitate healing. PMID- 15816218 TI - [Noise. Acoustic pollution in emergency rooms]. AB - In our current society, it is essential to consider acoustic contamination as a type of aggression which has a direct effect on the health and well-being of individuals. Therefore, it is necessary to establish a series of facts which will enable us to clarify the importance of the aforementioned problem. PMID- 15816219 TI - [The effects of noise on health]. AB - This article is adapted from the inaugural speech which Dr. Tolosa made to the Royal Academy of Medicine in the Balearic Islands during the 2003 academic year. In his speech, he analyzed how the presence of sound in our environment is such a common factor in our daily lives that we are hardly aware of its effects. Sound provides experiences as pleasant as listening to music or birdsongs, or sound enables oral communication among people; but together with these agreeable auditory perceptions, bothersome, even harmful, noise is also present in our lives; this noise can limit our relationships and affect our health in an irreversible manner. PMID- 15816220 TI - [Industrial nursing. Functions within the basic health unit]. AB - The Law of Prevention of Risks at Work marked an important point of flexion in the tie sanitary professions the field of the labor health. This work will mark the objective to locate to the Industrial Nursing (a company nursing assistant) in a context near the reality. For it, it will be equipped with legal content, and those articles of the Law of Prevention of Risks at Work will be mentioned and of the Regulations for Prevention Services, that establishes the guidelines of performance of the professionals of the Basic Unit of Health (Industrial Medicina and the company nursing assistant) Also will mention the objectives of the First draft of Law of the Reformation of Normative Marco of the Law of Prevention of Risks at Work, consequence of the alarming data of sinisterness and labor accidental. PMID- 15816221 TI - [Nurses who treat Parkinson patients: what do they know and what do they want to know?]. AB - To deal with Parkinson Disease, the education and the participation of the patient and his/her patient in the care of this disease are fundamental aspects. To know a patient's perceptions about his/her need for information, the patient's anxieties and the necessity to communicate with the medical team help to individualize the education provided for a patient. METHOD: a general qualitative descriptive study developed by means of semi-structured interviews. SAMPLE: 19 outpatients previously diagnosed with Parkinson's. The random method used was based on setting up interviews by the medical team by means of a computer. RESULTS: The majority of the patients felt their knowledge level was sufficient, although some search for more information and others showed a fear to learn more. The main source of information comes from their neurologists. The patients think that the information related to the symptoms and the drugs (for Parkinson's) is adequate, but this information is lacking regarding emotional necessities and other personal aspects. The patients think that communication is fundamental when dealing with a chronic disease, and that there is a need for individualized, very human, relationships where besides technical competence, patients hope for comprehension, security confidence and the opportunity for patients to express themselves. PMID- 15816222 TI - [LCT (lipocolloid technology) in lesions of venous etiology]. AB - This article reports on a spanish study to evaluate the effectiveness of dressings using a Technology Lipido-Colloid to treat skin lesions with "Urgotul". Has been included 18 of this 60 lesions were leg ulcers. We have decided to focus our analysis in these difficult to treat chronic wounds. This study noted better results in relation to the evolution of these lesions and in the quality of life for these patients who suffered these lesions in terms of pain control, skin deterioration or lesions in the same spot related to changing dressings, and an improvement in the skin nearby the lesion. PMID- 15816223 TI - [Semiautomatic external defibrillation: use inside a hospital]. AB - The electric rhythm which provokes the majority of the cases of sudden death is ventricular fibrillation, and the only effective measure to reestablish spontaneous circulation consists in administering a precocious electrical discharge, or defibrillation. Time is a fundamental factor since the possibilities tor survival are practically nil it defibrillation is applied 10 to 12 minutes after the patient suffers his/her attack. Therefore, in recent years, the use of semiautomatic external defibrillators has been promoted. These small, low-cost, easy to handle apparatuses are designed so that the first person who detects a cardio-respiratory arrest can defibrillate, whether or not that person is a trained medical technician. Bearing in mind that in our hospitals nurses are in close contact with patients 24 hours a day nurses become the most appropriate professional to utilize these devices once they have received minimum training and instruction in the use of semiautomatic external defibrillators which will qualify and authorize nurses to do so. PMID- 15816224 TI - Strategies and impacts of new drug introduction: hemophilia treatment. AB - This study examines the impact of the manufacturers' pricing strategy on the growing spending in the drug industry in the case of hemophilia treatment. Testable hypotheses of non-competitive pricing policies are posited to examine whether drug companies manipulate the prices of new drugs at each stage of market adoption. Then, we empirically examine our hypotheses using data from a regional hemophilia drug-purchasing consortium over an eight-year period. The key finding confirms that the pharmaceutical companies engage in non-competitive pricing behavior to maximize their profit given the limited competition in the market. The implications of the findings are considered. PMID- 15816225 TI - The effectiveness of wellness programs as a strategy for cost containment in acute care hospitals. AB - This study examines the effectiveness of hospital-based wellness programs in lowering both the acuity of illness of patients and the total expenses of acute care hospitals from a strategic management perspective. The subjects for this cross-sectional study were 164 community hospitals in 27 urban areas of Texas. The findings show that, after controlling for size, the number of wellness programs was significantly and negatively related to both the acuity of illness and total expenses. Further, the number of wellness programs offered did not vary significantly by type of ownership. The study concludes that reimbursement policies designed to provide financial incentives to promote wellness have been effective and suggests future directions for the evolution of health care management. PMID- 15816226 TI - Length of hospital stay of Medicare patients in the post-prospective-payment system era. AB - This study examines factors determining hospital length of stay (LOS) for Medicare patients in the post-prospective-payment-system (PPS) era according to type of ownership, geographical region, bed size, and financial performance. The study design includes a number of economic and financial variables. In the empirical findings, the following factors are significant in determining hospital LOS of Medicare patients: hospital size, type of ownership, profitability, teaching status, and competition. Other studies have examined factors influencing length of hospitalization; however, none of these studies investigates the effect on LOS of a change in the type of hospital ownership. Furthermore, this study uses more recent and comprehensive data than in the current literature. PMID- 15816227 TI - The effects of knowledge management on surgeon behavior. AB - Knowledge management is an important process for health care researchers and administrators. The way we manage and transfer knowledge in an organization can have a substantial impact on behavior and performance. In this article, we examine the behavioral effects of transferring performance-efficiency knowledge to a group of hospital-based surgeons. We observe the way the knowledge transfer impacts their sense of professional accountability and practice patterns for a limited set of diagnoses. We defined performance efficiency for a surgeon as the deviation from expected average length of inpatient hospital stay, and from expected average hospital charges (adjusted for risk and outcomes) for three of the most frequently performed and most costly surgical procedures in our subject hospital. We communicated knowledge of their performance efficiency to the group of hospital-based surgeons, along with benchmarked professional best practices, and included an identification of dimensions where performance could be improved. We then measured and compared their performance efficiency one year later. We did observe differences in performance efficiency, but not in consistent directions, and not in statistically significant magnitudes. Also, surgeons who initially had low levels of efficiency continued to have low levels of efficiency one year later. Within a professional accountability system, transfer of performance efficiency knowledge alone did not provide sufficient motivation to induce consistent, significant change in practice behaviors among the group of surgeons. We conclude that medical opinion leaders and individualized strategies for surgeon motivation may have greater promise for improving performance efficiency if linked to the knowledge transfer system. PMID- 15816228 TI - The impact of DRG-based payment systems on quality of health care in OECD countries. AB - Ever since DRG-based payment systems were first introduced in the United States in 1983, the medical community has expressed concern about the potential impact of these price control systems on the quality of care. Several research studies have examined the impact of DRG-based payment systems on the quality of care within a single state in the United States, or within a specific country. We have not identified any attempts in the literature to examine the impact of DRG-based payment systems on the quality of health care across different countries. In this article we contribute to the debate by (1) providing a unique identification of DRG adoption status for each of 35 countries, (2) refining an international case mix index, and (3) applying it to examine whether DRG-based payments impact the quality of health care across national and cultural boundaries. We find some evidence for Organization for Economic Cooperation and Development countries that, compared with non-adopters, adoption of DRG-based payment systems is associated with faster hospital case mix increases and slower quality gains with respect to patient mortality from surgical and medical misadventures. PMID- 15816229 TI - Consumer responses to a pharmacy benefit report card. AB - As pharmacy benefits become an increasingly important component of rising health care costs, report cards critically need to be developed to inform consumers about the availability of their medications, what these medications cost them, and additional pharmacy benefit services that can enhance the management of their disease. This study is a preliminary step for developing such a card and assessing its impact on health plan enrollment choices in a small employee sample. Findings indicate that while individuals generally had positive comments about the pharmacy benefit report card, the card had little effect on their health plan enrollment decisions. The higher the prescription use reported by individuals, more likely they were to report that the card influenced their health plan enrollment decisions. The pharmacy benefit report card could be an useful tool for specific populations such as those with an increased need for medication. PMID- 15816230 TI - The influence of hospital integration on hospital financial performance. AB - A clinical and functional integration strategy has a positive influence on increasing hospital revenue, and a solely functional integration strategy has a negative influence on increasing hospital expenses. Functional integration and clinical/functional integration strategies have a positive influence on hospital profit and the overall operations of the hospital. The mechanism of influence differs, however, based on the strategy used. Clinical/functional strategy has an impact on increasing hospital revenue, while functional integration strategy has an impact on reducing hospital expenses. Overall, the study shows that a functional integration strategy is more profitable than a clinical/functional integration strategy. PMID- 15816231 TI - Angiosarcoma replacing bone marrow--a case report. AB - BACKGROUND: Clinical syndromes caused by marrow replacement by neoplastic cells are usually proven to arise from cells of hematopoetic or carcinomatous lineage. Diffuse marrow replacement by mesodermal origin sarcoma is rare in adults, and angiosarcoma replacing bone marrow is a very rare neoplasm. METHODS: A case of bone marrow replacement by angiosarcoma in a 54-year-old African American man is described with a review of pertinent literature. The unique histopathologic features of this phenomenon are also described. RESULTS: Histopathologic exam of trephine bone marrow biopsy revealed replacement of marrow spaces by a proliferating highly vascular malignant neoplasm with features of high-grade angiosarcoma. CONCLUSION: Few reports in the literature describe angiosarcomatosis of the bone marrow. It is important to recognize this unusual presentation to avoid confusion with other more common bone marrow replacing entities and to promote recognition that more efficacious treatment is needed. PMID- 15816232 TI - Just another pneumonia? PMID- 15816233 TI - Notes of a policy watcher: the Medicare Drug Plan. AB - The Medicare Modernization Act, signed into law by President Bush in December 2003, provides for new prescription drug benefits to those enrolled in Medicare. This new benefit will provide many seniors with improved access to medications and should thereby improve outcomes of care. The benefit is complex, though, and we are currently in a transition period. The text discusses current benefits and drawbacks of the Medicare Modernization Act Drug Plans as well as what is planned for 2006. PMID- 15816234 TI - "Goldberger's War" in Mississippi: the 1915 Pellagra Experiment at Rankin State Farm and a new book. PMID- 15816235 TI - Expert witness. PMID- 15816236 TI - Mississippi needs more doctor mayors: "get out, have fun, and make a difference". PMID- 15816237 TI - The rock and the hard place. PMID- 15816238 TI - Statistics show that Mississippi health care providers are improving care. PMID- 15816239 TI - Building better bodies. A MSMAA state-wide health project. PMID- 15816240 TI - USDA choice spam: an epiphany. PMID- 15816241 TI - Battle over unlicensed medication administration continues. PMID- 15816242 TI - Gait abnormalities are too hard to evaluate, so why bother? PMID- 15816243 TI - The singular benefits of bathing. PMID- 15816244 TI - Stroke prevention. PMID- 15816245 TI - First steps: a program for medical students to teach high school students about breastfeeding. PMID- 15816246 TI - Ultra-rapid aortic stenosis progression in end stage renal disease. PMID- 15816247 TI - Images in medicine. Left humerus and intertrochanteric fracture in an elderly man. PMID- 15816248 TI - Miriam Hospital and Rhode Island Hospital morbidity and mortality conferences, case of Lemierre syndrome. PMID- 15816249 TI - The impact of poverty on prevention practices and health status among persons with asthma. PMID- 15816250 TI - The "Low Carb Craze" and current fad diets. PMID- 15816251 TI - Stem cells in medicine: designing the clinical research. PMID- 15816252 TI - What motivates idealistic young people as they plan their careers? PMID- 15816253 TI - Faculty matters. Diane Hamilton. PMID- 15816254 TI - Perspectives on improving nursing's public image. 1980. PMID- 15816255 TI - The Texas model of differentiated entry-level competencies of graduates of nursing programs. AB - The Board of Nurse Examiners and the Board of Vocational Nurse Examiners for the State of Texas updated the state's competency-based education model in collaboration with stakeholders in nursing service, state nursing organizations, and the deans and directors of the state's 82 board-accredited professional nursing programs. Based on this collaborative effort, 14 differentiated entry level competencies with associated knowledge and clinical behaviors/judgments were identified that set standards for graduates of vocational, diploma, associate, and baccalaureate degree programs. PMID- 15816256 TI - Preventing violence in an academic setting: one school of nursing's approach. AB - Workplace violence occurs in many different contexts, including academic settings. However, the types of violence that make headlines are less common than the everyday behaviors that, left unchecked, may lead to extreme situations. This article describes how a school of nursing developed a program of workplace violence prevention, intervention, and response. Descriptions of the program, definitions, and workplace violence resources are included. PMID- 15816257 TI - Developing the research pipeline: increasing minority nursing research opportunities. AB - The need to increase the number of minority nurses in policy and leadership positions in health care administration, academia, and research has been acknowledged. Limited academic and research training opportunities are available specifically designed to develop a cadre of minority nurse scientists to conduct the research needed to ultimately reduce health disparities within racial and ethnic minority populations. This article describes a collaborative approach to research career development at the doctoral level. PMID- 15816258 TI - How can I help you? how can you help me? Transforming nursing education through partnerships. PMID- 15816259 TI - People of the year: bloggers. PMID- 15816260 TI - Substantive innovation in nursing education: shifting the emphasis from content coverage to student learning. PMID- 15816261 TI - Infections linked to personal appearance workers. PMID- 15816262 TI - Cancer incidence and mortality in Grenada 1990-2000. AB - This paper summarizes and discusses the available cancer incidence (1996-2000) and mortality data (1990-2000) for the tri-island Caribbean nation of Grenada, Carriacou and Petit Martinique. Data for the analysis came from three sources: the Grenada Department of Statistics, the histopathology specimen books from St George's General Hospital and the Death Registry of the Ministry of Health, Grenada. The age-standardized rates (ASR) per 100 000 for all cancer sites combined were 170.2 in females and 158.2 in males. The four most frequent diagnoses (ASR) by cancer site in females were cervix (60.7), breast (49.1), uterus (28.4) and skin (13.3); and among males, prostate (61.4), bladder (16.3), skin (19.3) and stomach (10). Age-standardized mortality rates per 100 000 for all cancer sites combined were 105.4 in females and 165 in males. The four most frequent cancer associated mortalities (ASR) in females were breast (17.9), uterus (11.2), colon (10.3) and cervix (9.7); and among males, prostate (53.6), lung (18.7), stomach (14.5) and colon (10.9). This study found statistically significant spatial trends for overall cancer mortality and temporal trends in incidence and mortality rates for prostate and for incidence rates of stomach cancer. These rates are compared with those from other areas in the Caribbean and the United States of America and encourage efforts to establish a cancer registry in Grenada. PMID- 15816263 TI - Antibiotic sensitivity of Helicobacter pylori in Jamaica. AB - Helicobacter pylori infection is one of the commonest chronic infections worldwide. Eradication regimes usually contain two antibiotics, however resistance is increasing and this decreases treatment success. This study reports on the sensitivity and resistance of H pylori to several antibiotics in patients undergoing upper gastrointestinal endoscopy in Jamaica. The rapid urease test (CLO) was positive in 128 (38%) of 336 patients. Fifty patients (39%; 50/128) with positive CLO tests had positive cultures for H pylori. Two-thirds (32/48) of islolates were sensitive to metronidazole and one-third (16/48) were resistant. Ninety-seven per cent of isolates (31/32) were sensitive to erythromycin. The sensitivity for clarithromycin was 92% (11/12) with one isolate (8%) resistant. All strains of H pylori (48/48) were sensitive to ampicillin and amoxicillin - clavulanate. Metronidazole resistance is present in one-third of H pylori isolates and resistance to macrolides is relatively low in Jamaican patients. It is important to monitor antibiotic resistance in order to provide clinicians with data on the most appropriate and cost effective eradication regimes for H pylori. PMID- 15816264 TI - Surgical management of perforated duodenal ulcer: the changing scene. AB - To determine the management of perforated duodenal ulcer at the University Hospital of the West Indies (UHWI) in this era of Helicobacter pylori, the medical records of all patients seen at the UHWI during the period July 1997 to June 2002 with an intra-operative diagnosis of perforated peptic ulcer were reviewed The records were analyzed for the following: age, gender, duration of symptoms, non-steroidal anti-inflammatory drug (NSAID) use, smoking status, operative repair duration of hospitalization, Helicobacter pylori status and medical therapy, peri-operative complications, mortality and recurrence. Ninety per cent of the cases were males. All females in whom perforation occurred were age 50 years and older compared to males where 58% of cases presented before age 50 years. Perforations in acute ulcers occurred in 80% of cases. The majority of patients were male smokers. Non-steroidal anti-inflammatory drug use was also an important risk factor in elderly females. Simple surgical closure and standard triple therapy antibiotics to eradicate Helicobacter pylori was the most common treatment offered. Mortality was one per cent and follow-up poor but 11% of patients had documented recurrent peptic ulceration. In this study population, perforated duodenal ulcer occured overwhelmingly in males less than 50 years of age. There is a trend towards exclusive simple surgical closure and H pylori eradication at the UHWI for patients with perforated duodenal ulcer but this needs to be supported by documentation of H pylori prevalence in the population of patients presenting with perforated peptic ulcers. PMID- 15816265 TI - Civilian rectal trauma: the surgical challenge. AB - Using the Department of Surgery, Radiology, Anaesthesia and Intensive Care's operative database and information from the Trauma Registry for patients presenting after 1998, a retrospective study of patients seen between the period 1992 and 2002 was done at the University Hospital of the West Indies with the objective of determining the treatment and outcome of patients with a diagnosis of rectal trauma. Over the 10-year period, 45 patients were seen with this diagnosis. Eighty-two per cent of the cases were males, with a mean age of 29.8 years (range 16-70 years) while the eight female patients had a mean age of 36.8 years. Low velocity gunshot wounds accounted for 64% of the entire group and for 78% in males. Six of the eight cases seen in females were iatrogenic occurring during gynaecological operations. Sixty-three per cent of rectal injuries were associated with other injuries with the genitourinary system most commonly involved The majority (83%) were diagnosed preoperatively by visualization of the rectal wound or the presence of blood on proctosigmoidoscopy. All the patients had peri-operative antibiotics and diversion with a proximal sigmoid colostomy. Fifteen per cent of cases had presacral drain insertion. Distal rectal washout was not used. There were no deaths. Seventy-three per cent of patients had closure during the period under review at an average time of 8.5 months after initial surgery. The other 27% after an average of 23 months did not have documented closure. The mainstay of treatment for civilian rectal trauma remains diverting sigmoid loop colostomy, despite its morbidity, and peri-operative antibiotics. PMID- 15816266 TI - Assessment of risk for type 2 diabetes mellitus in a Caribbean population with high diabetes-related morbidity. AB - Diabetes mellitus is a major cause of morbidity in Trinidad and Tobago. Screening programmes are not incorporated in the health sector and the population at risk remains unaware of the benefits of screening. We investigated the risk of developing Type 2 diabetes mellitus in office workers with one risk factor. Participants were randomly selected from the urban corporate sector in Port of Spain. Fasting capillary blood glucose and the American Diabetes Association (ADA) questionnaire for major diabetes risk factors were used to assess risk. Student pharmacists approached 482 persons, of whom 317 consented to participate (66% response rate). There were 101 (32%) men and 216 (68%) women, 37 (39%) were of African ancestry and 28% each were of East Indian and mixed ancestry. Family history was positive in 54%. Thirty per cent (95) of the volunteers were at risk of developing Type 2 diabetes mellitus (41 men; 54 women). Based on the ADA questionnaire, 82% (78) of volunteers were at high risk for developing Type 2 diabetes mellitus. The ADA risk test and Impaired Fasting Glucose were both positive in 13 (14%) volunteers. In subjects at risk, Body Mass Index (BMI) was > 25 kg/m2 in 74% (78) and the waist/hip ratio was 0.85. Approximately 30% of office staff was at risk of developing diabetes mellitus. The ADA questionnaire is a useful non-invasive measure which pharmacists can use to assess risk for Type 2 diabetes mellitus. The glucometer can be used for risk assessment providing that it is associated with a quality assurance programme and that diagnosis is confirmed with laboratory testing. PMID- 15816267 TI - Postprandial glucose control in Type 2 diabetic patients visiting two different primary care clinics in Trinidad, West Indies. AB - The aim of this study was to determine the differences in postprandial glucose levels between Type 2 diabetic patients visiting two different primary care clinics after ingestion of three carbohydrate test foods. Thirty-eight Type 2 diabetic patients visiting the Chaguanas and Arima Lifestyle Disease Clinics and 27 healthy non-diabetic subjects were studied on three different occasions, seven days apart, after an overnight fast. On each day of the study, anthropometric indices were measured and after collecting fasting blood samples, subjects randomly consumed three pre-selected carbohydrate test foods: bread, roti or rice within 10 minutes and water taken as wished. Subsequently, seven millimetres of venous blood samples were collected at 60, 90, 120 and 150 minutes for insulin, glucose and lipid determinations. Age, body weight, body mass index, waist and hip circumferences did not differ between the healthy subjects and diabetic patients or between patients of the two clinics (p > 0.05). The mean fasting and 2.5-hour postprandial glucose values for the patients visiting the Chaguanas clinic were higher, after consuming at least one test food, than those of patients visiting the Arima clinic (p < 0.05). The mean HbA1c, and 2.5-hour glucose levels for the diabetic patients visiting either of the two clinics exceeded the internationally recommended cut-off values; this is worst in patients visiting the Chaguanas clinic. It is suggested that primary care physicians should start by targeting glycaemic control based on international standards while a more aggressive diabetes dietary education should be reinforced at the clinics, especially Chaguanas. PMID- 15816268 TI - The practice of polypharmacy involving herbal and prescription medicines in the treatment of diabetes mellitus, hypertension and gastrointestinal disorders in Jamaica. AB - The interference in the metabolism of a drug by another drug, food or other foreign chemical is commonly observed and often leads to clinically significant adverse drug reactions. In Jamaica, there is an extensive use of natural products as medicines, although there is little information on whether natural medicines are used along with prescription medicines, which might increase the likelihood of drug adversities. This current pilot survey was initiated to gain information on the prevalence of such polypharmacy practices. Two concurrent surveys were carried out in Kingston (an urban parish) and Clarendon (a rural parish) in 743 patients above age 14 years with diabetes mellitus, hypertension and gastrointestinal disorders of persons who visited health centres and pharmacies. Patrons visiting these places at various times of the day were informed of the nature of the questionnaire and willing participants with the above disease conditions and who were on prescription medicines were included in the survey. The results indicated that 80% of the respondents reported combining natural products in their treatment along with prescription medicines, with only 13% of those patients informing their physicians of such practices. Such polypharmacy practices were independent of the type of disease among both males and females and was statistically most prevalent (p < 0.001) in the 47-57-year age group. There was a significant association (p < 0.001) between such treatment practices and place of residence with 92% of the rural community engaging in polypharmacy but 70% of the urban did likewise. Therefore, being aware of the prevalence of multiple therapy practices would be especially useful when designing a pharmaco vigilance system. PMID- 15816269 TI - Depo medroxyprogesterone acetate (DMPA) and combined oral contraceptives and cervical carcinoma in-situ in women aged 50 years and under. AB - Most low-resource settings depend on hormonal contraceptives for their family planning programmes and cervical cancer occurs in higher frequency in these populations. To determine whether hormonal contraception use increases cervical carcinoma in-situ (CIS) risk, a case-control study was conducted in the Kingston and St Andrew Corporate area of Jamaica, using 119 cases from the Jamaica Tumour Registry and 304 population controls matched on year of Papanicolaou (Pap) smear and clinic where Pap smear was obtained. While CIS cases were more likely to have 'ever used' combined oral contraceptives (COC) (OR = 1.4, 95% CI: 0.8, 2.5), depo medroxyprogesterone acetate (DMPA) use was similar. Compared to women who never used hormonal contraceptives, the risk of CIS was elevated in: women who had used COCs five years or more (OR = 2.1, 95% CI: 1.0, 4.6), women who first used COC for less than 10 years prior to the interview (OR = 1.8, 95% CI: 0.9, 3.7) and women who were 18 to 24 years old when they first used COCs (OR = 1.8, 95% CI: 0.9, 3.4). Similarly, compared to women who never used DMPA, the risk of CIS was elevated in: women using DMPA five years or more (OR = 1.9, 95% CI: 0.7, 4.8), women reporting use within a year prior to interview (OR = 2.8, 95% CI: 0.7, 10.7) and women who initiated use of DMPA when they were 20 and 24 years old (OR = 1.4, 95% CI: 0.7, 3.1). These results suggest that if hormonal contraceptive use confers any risk of CIS, it is confined to long-term users. Increased risk in some groups, however, warrant further study. PMID- 15816270 TI - Isolation and characterization of microorganisms from instruments used by pedicurists operating within Lagos metropolis, Nigeria. AB - Eight bacterial and five fungal species were isolated from swab samples taken from instruments used by pedicurists operating at three different sites in Lagos, Nigeria. The bacterial isolates included Micrococcus luteus, Micrococcus roseus, Staphylococcus epidermidis, Staphylococcus aureus, Hafnia spp, Shigella spp, Bacillus subtilis and Bacillus spp. The five fungal isolates were identified as Aspergillus niger, Aspergillus flavus, Mucor spp, Trichophyton spp and Candida albicans. The presence of these microorganisms, some of which are pathogenic, is an indication that pedicurists could be contributing towards the spread of skin and nail infections within the Lagos metropolis. PMID- 15816271 TI - An unusual presentation of group B streptococcal sepsis. AB - This paper describes the devastating outcome of a neonate who presented with severe late onset group B beta-haemolytic streptococcal (GBS) disease. There were extensive infarcts of the brain and gangrene of the toes. The purpose of this report is to alert healthcare workers of the unusual presentation and that fatal late onset group B beta-haemolytic streptococcal disease may occur despite early and effective management. PMID- 15816272 TI - Community-based prevention of substance abuse. AB - Substance abuse is pervasive in a number of countries throughout the world, placing an enormous burden and strain on their healthcare systems. The relationship between crime and illicit drug use increases the significance of the problem facing countries worldwide. Jamaica has not only become a trans-shipment point for cocaine from South America, but recent Drug Enforcement Agency figures show Jamaica being responsible for 30% of cocaine trafficking into the United States of America. Though all segments of society are involved, disadvantaged groups are disproportionately affected. Substance use and abuse among adolescents is of particular concern. Supply reduction approaches have not been effective enough in reducing the prevalence of substance use and abuse in many countries throughout the world. Substance abuse prevention research over 25 years has led to the identification of a number of factors differentiating substance abusers from others. The presence of risk factors and the absence of protective facts contribute to an increase potential for drug abuse. The impact that these factors have is also determined by the level of psychological and social development of an individual. The presence of risk factors as well as the absence of protective factors leads to increased "exposure opportunity", exposure to a potential substance of abuse being closely associated with the potential for initial drug use, transition from initiation to regular use and subsequently dependence. One method of reducing "exposure opportunity" is community prevention. The basic aim of community prevention programmes is to decrease the rate of drug use in a population by enhancing protective factors and reversing or reducing risk factors. PMID- 15816273 TI - Image and diagnosis. Huge abdominal aortic aneurysm with intra-arterial clot with wall calcification. PMID- 15816274 TI - William Harvey and his discovery of the circulation of the blood. PMID- 15816275 TI - Krukenberg tumour from an appendiceal carcinoma presenting as a primary ovarian tumour. PMID- 15816276 TI - Traumatic gastropleural fistula. PMID- 15816277 TI - Everyday heroes and heroines: real-life nursing examples. PMID- 15816278 TI - Who is the California Nurses Association and why are they coming to North Carolina? PMID- 15816279 TI - What nursing shortage? PMID- 15816280 TI - Aged care nurses reject QH proposal. PMID- 15816281 TI - Howard government attack on unions begins. PMID- 15816282 TI - HIV/AIDS program in Ethiopia. PMID- 15816283 TI - The Governator takes on Californian nurses. PMID- 15816284 TI - Patient fact sheet. Dental health concerns for infants and toddlers. PMID- 15816285 TI - For clearer judgment. PMID- 15816286 TI - The struggles of a young dentist seeking respect. PMID- 15816287 TI - ADA working to bring volunteer dentists to Alaska. PMID- 15816288 TI - What should I do with my dental records when I retire? PMID- 15816289 TI - Top five opportunities to improve your profit. PMID- 15816291 TI - The business of dentistry: talking points for speakers on dental careers. PMID- 15816290 TI - New CareCredit software feature helps case acceptance. PMID- 15816292 TI - Dr. Bob Russell: "My mission is to serve my community"! PMID- 15816293 TI - Your dental team's standards communicate quality. PMID- 15816294 TI - Do implantable cardioverter-defibrillators reduce the risk of death? PMID- 15816295 TI - A qualitative study of insurers' coverage for mifepristone-induced abortion. AB - In this qualitative evaluation of 14 U.S. health insurers' coverage policies concerning mifepristone-induced medical abortion, the findings suggest that a significant proportion of health plans do not include evidence-based clinical information obtained from recent research, data, and practice. This omission may hinder the adoption of new, evidence-based protocols for medical abortion by physicians. A model coverage policy that contains comprehensive clinical information regarding appropriate clinical regimens for mifepristone is provided as an example for insurers and others in the field. PMID- 15816296 TI - Disease management challenges in rural settings. PMID- 15816297 TI - Chronic disease management in rural and underserved populations: innovation and system improvement help lead to success. AB - Successful implementation of disease management (DM) is based on the ability of an organization to overcome a variety of barriers to deliver timely, appropriate care of chronic illnesses. Such programs initiate DM services to patient populations while initiating self-management education among medication-resistant patients who are chronically ill. Despite formidable challenges, rural health care providers have been successful in initiating DM programs and have discovered several ways in which these programs benefit their organizations. This research reports on six DM programs that serve large rural and underserved populations and have demonstrated that DM can be successfully implemented in such areas. PMID- 15816298 TI - A strategy for affordable access to Medicare drug coverage. PMID- 15816299 TI - The use of topical retinoids for inflammatory acne: a review of the pharmacoeconomic considerations. AB - Few pharmacoeconomic studies exist regarding the use of topical retinoids for inflammatory acne vulgaris. Those pharmacoeconomic studies that do exist predictably involve new entrants (topical tazarotene in various formulations). The use of topical retinoids in combination with antimicrobial agents constitutes cost-effective therapy for inflammatory acne. The growing U.S. trend toward use of isotretinoin for less-severe forms of acne instead of topical retinoid-based combination therapy is problematic from clinical and cost perspectives. More pharmacoeconomic studies that consider the importance of compliance are necessary for clinicians and managed care professionals to make informed choices about the treatment of inflammatory acne. PMID- 15816300 TI - Utilization of health care services among the near-elderly: a comparison of managed care and fee-for-service enrollees. AB - This article compares and contrasts the use of health care services among near elderly adults (55-64 yr) across HMOs, PPOs, and fee-for-service (FFS) plans. Statistical tests are used to examine possible selection bias by type of plan [corrected] Multistage regression methods are implemented whenever the test results indicate the presence of selection bias. The findings show that compared with FFS plans, HMOs and PPOs are associated with a greater use of outpatient services and preventive services among adults in this age range. The rate of hospital admissions and the length of hospital stay are found to be similar across these types of plans. PMID- 15816301 TI - [Pharmacotherapeutic trends at the beginning of the millennium in Hungary: pharmacotherapy of bipolar patients. Part II]. AB - The aim of the study was to evaluate the pharmacotherapeutical trends in the treatment of bipolar outpatients in Hungary. The first part of the paper included the basic data and this second part discusses more detailed results from more aspects. Hungarian and international trends were compared. The investigation of the number of simultaneously given different drugs showed that bipolar II patients take the largest number of psychotropics within the bipolar spectrum, and patients with mania or depression as actual condition. The use of mood stabilizers is frequent (80%), and it is in line with the international data, but the rate of carbamazepine (25% as prophylactic drug) is higher in Hungary than in other countries. The rate of the patients on permanent anxiolytics is higher than expected. The gender differences are not significant; women are given more antidepressants and anxiolytics than men, but the prevalence of actual depressive phases is higher in the female than in the male population. There were some significant differences in the therapeutic trends (use of mood stabilizers and antipsychotics) among the Hungarian regions and between the different types of institutions. The international comparison showed that the rate of Hungarian bipolar patients on lithium was significantly lower than in other countries. Although studies with old and new drugs as well as the progress of clinical pharmacology influence everyday practice constantly, the results of this investigation provide some basic data for the trials in the future. PMID- 15816302 TI - [Costs related to change to amisulpride in patients suffering from schizophrenia]. AB - OBJECTIVES: The present research studied the cost effects of converting patients suffering from schizophrenia to the use of amisulpride, in order to learn whether the relatively higher medicine costs were compensated for under the Hungarian economic and financing conditions. METHODS: We analysed and compared costs having occurred before and after conversion in the case of 76 patients suffering from schizophrenia who got amisulpride instead of other typical or atypical antipsychotics in hospital. The analysis adhered to the methodology introduced by Agnes Rupp. RESULTS: In conformity with earlier investigations in Hungary performed with atypical antipsychotics, amisulpride has also proved its cost neutrality under local economic and financing conditions. Namely, higher disbursements for medicaments are compensated for by productivity increase, indicative of amisulpride's effectiveness, implying decreased economic (and other) burdens to family members. Te remarkable cost ratio improvement in case of patients defined as therapy resistant has played an important role in cost neutrality. CONCLUSIONS: Amisulpride is an effective antipsychotic which can be prescribed without increasing costs, and in case of therapy resistant patients, it appears to have significant cost sparing effects. PMID- 15816303 TI - [Psychiatric symptoms in general medical hospital units--assessment of the need for psychiatric consultation-liaison in Hungary]. AB - AIM: Psychiatric symptoms and disorders are a serious problem area, absorbing high health service provision capacities. Extensive research work is done internationally relating to the morbidity of psychiatric disorders in general medical departments, but no Hungarian data have been available so far. In the present study, general psychiatric symptoms were detected in Hungarian general hospitals, with special regard to mood disorders. METHODS: A self-assessment based questionnaire survey was performed, covering 2,444 patients treated in general medical departments. The prevalence of depressive symptoms was measured by the Beck Depression Inventory. The use of anti-depressants was also checked. RESULTS: On the basis of the questionnaire screening, 46.6% of patients showed a pathological score. Depressive symptoms were found primarily in the departments of chronic diseases. The use of antidepressants showed a very low frequency, in spite of the high prevalence of depressive symptoms. The use of anxiolytics was relatively high in depressive states. CONCLUSION: The high frequency of mood disorders is not surprising in somatic medicine, but it is mainly found among chronic patients. However, their adequate pharmacotherapy is insufficient. On the basis of these data, the improvement of the consultation-liaison psychiatry is needed. PMID- 15816304 TI - [Neurophenomenological analysis of the therapeutic relationship in ritual healing]. AB - In this essay we examine some phenomena of community rituals, especially healing ceremonials, which are considered neurobiologically mediated, complex forms of attachment. Recent studies in medical anthropology have pointed out that the ritual therapeutic experience relies on the patients' own healing processes by means of various altered states of consciousness that healers are able to control. Ritual trance invariably occurs in social context, and the healer's personality and the expectations of the community are profoundly involved in the induction of altered states of consciousness. Trance state is regarded as a result of the mobilisation of endogenous opiates, as an activation of the organism's defensive mechanisms in face of the stress of ceremonial. On the other hand, there is a growing body of evidence that opiate mechanisms are involved in social behaviour as well, especially in symbiotic bonds. It is suggested that this is the neurobiological reason why attachment facilitates trance induction. The homeostatic role of social relationships as physiological regulators is also discussed. PMID- 15816305 TI - [Monoaminergic activity acts on the visual evoked potentials]. AB - Animal experiments show that pyramidal cells obtain strong noradrenergic innervation in the primary visual cortex. Its neurotransmitter, norepinephrine, has strong inhibitory-hyperpolarizing effect on these cells. Because pyramidal cells are known to be the main generator of cortical bioelectrical activity, we supposed correlation between monoaminergic activity and evoked potential. In the present study this hypothesis was tested using 31 healthy volunteers. As pyramidal cells possess contrast and orientation specificity in the visual cortex, a checkerboard pattern was used for stimulation, at four light intensities. For statistical analysis, the relative peak amplitudes (CI-CII, CII CIII) and peak latencies (L1, L2, L3) of VEP components were used. To characterise noradrenergic activity, platelet monoamine oxidase and plasma dopamine-beta-hydroxylase were determined from blood samples. We also measured the plasma cortisol level, which is also a good marker of vegetative activity. We found a significant negative correlation between MAO activity and the amplitude of VEP; a strong negative correlation between CS level and L1 latency, and a significant positive correlation between DBH level and L1 latency. Our results prove not only the close correlation between the MAO activity and the bioelectrical characteristics of the visual cortex, but also suggest that VEP could be a good noninvasive tool for the characterisation of central monoaminergic activity. PMID- 15816306 TI - [Neurochemical background of the premenstrual syndrome: the role of the serotonergic system]. AB - Current research focusing on the biological background of premenstrual syndrome suggests that both psychological and physiological factors exert their effect by means of the dysregulation of the serotonergic function. The results of several studies point to a variation in the function of the serotonergic system throughout the cycle, which is related to the plasma concentration of the gonadal steroid hormones. Certain parameters of serotonergic function show different values in the different phases of the cycle. The rise in oestrogen concentration is followed by an increase in the 5HT2A receptor expression in brain areas responsible for mood, emotion, cognition and neuroendocrine regulation. The role of serotonin is further supported by its role in depression, anxiety, aggression as well as the efficiency of serotonergic agents in the treatment of premenstrual syndrome. In our article we review the main theories concerning the biological background of the premenstrual syndrome, and we outline in detail the role of the serotonergic system. We review the research on the relationship between serotonergic function and gonadal steroid levels as well as the latest results concerning the state- and trait-like parameters of the serotonergic system. The studies reviewed provide evidence for the relationship between the gonadal steroid function and the serotonergic system. Further research is necessary in order to determine why the cycle-dependent variation in the function of the serotonergic system leads to the development of premenstrual syndrome only in vulnerable women. PMID- 15816307 TI - The histaminergic system of the brain: its roles in arousal and autonomic regulation. PMID- 15816308 TI - Warfarin protocol yields big reduction in adverse drug events. PMID- 15816309 TI - Long Island Health System standardizes care across 16 hospitals. AB - With 16 hospitals, there is a lot of potential for variation within the North Shore-Long Island Jewish Health System. But patients are assured that when they walk into one North Shore-LIJ hospital for treatment, the care they receive will be just as good as that provided in the system's other facilities. PMID- 15816310 TI - Wound care center improves treatment, increases revenues. AB - Patients are traveling to the Shawnee Mission Medical Center in Shawnee Mission, KS, for help with a painful and stubborn problem: wound care. Surgical openings that have failed to heal properly, along with wounds caused by infections, trauma, and diabetes, are treated at the wound care center with more than a 90% success rate. PMID- 15816311 TI - Benchmarking project spurs better guideline adherence. AB - A relatively simple benchmarking method has paid off in improved quality of care for Alabama diabetes patients. PMID- 15816312 TI - Quality, patient safety recognized in incentive plans, survey finds. AB - In an effort to improve the quality of care, healthcare organizations are likely to include quality performance targets in their incentive plans, according to a survey. PMID- 15816313 TI - The adolescent behavioral repertoire: its latent structure in the PACARDO region of Latin America. AB - In this study, the authors probed the latent structure of the adolescent behavioral repertoire (ABR) and estimated its sociodemographic correlates. The authors drew a nationally representative sample of 12,797 school-attending youth from the 7 countries in the PACARDO region of Latin America: Panama, Guatemala, El Salvador, Honduras, Nicaragua, Costa Rica, and the Dominican Republic. On the basis of the Behavioral Repertoire Self Report scale (Johanson, Duffy, and Anthony, 1996), the authors identified 5 primary dimensions, including religious behaviors, socializing, sports, home-based activities, and gender socialization activities. The authors found that the levels of involvement in these dimensions of the ABR varied across sociodemographic characteristics. The observed multidimensional structure of the ABR sets the stage for future research on adolescent health in relation to these behaviors and activities. PMID- 15816314 TI - Impact of two types of expectancy on recovery from total knee replacement surgery (TKR) in adults with osteoarthritis. AB - This study examines the impact of 2 classes of psychological variables- expectancies for the future and self-efficacy--on recovery of function and reduction in symptomatology among individuals seeking total knee replacement surgery (TKR). The authors collected outcome measures (SF-36, WOMAC, Clinician assessments) prior to surgery, 4-6 weeks after surgery, and 6 months postsurgery for surgery patients and controls. Linear trend analyses by group gauged the impact of the surgical intervention on recovery, revealing significant improvements over time on physical health outcomes for the surgery patients not attained by controls. In addition, two classes of psychosocial variables- expectancies and efficacy beliefs--predicted the extent of gains in physical and mental health outcomes among those patients who elected to have surgery. When initial status was controlled, between 9 and 13 percent of the variance in TKR outcomes was due to expectations and efficacy. These results point to the importance of psychosocial variables in recovery of function and reduced symptoms after knee surgery. PMID- 15816315 TI - Physical activity and quality of life: assessing the influence of activity frequency, intensity, volume, and motives. AB - In the present study, the authors investigated the impact of exercise frequency, intensity, and volume along with exercise motives on quality of life (QOL) reports. The authors assessed exercise habits with the Godin Leisure Time Activity Scale and measured exercise motives with the Reasons for Exercise Inventory. The Quality of Life Inventory assessed satisfaction in 16 domains including health, work, and recreation. High-frequency exercisers reported significantly higher health, helping, and community-related QOL than those who exercised less frequently. The authors noted significantly higher health-related QOL in the heavy volume group compared with the other volume groups. Multiple regression tests revealed that activity intensity and exercise motives significantly predicted QOL reports. The strongest bivariate correlations with QOL existed for mild activity and exercising for fitness and health reasons. Thus, high-frequency activity of mild intensity that produces high kcal utilization and is performed to improve health and fitness has the strongest influence on QOL reports. PMID- 15816316 TI - Effects of smoking, distraction, and gender on pain perception. AB - Whereas research on the antinociceptive effects of cigarette smoking has produced inconsistent findings, certain aspects of attentional processing repeatedly have been found to have an impact on pain perception. In an effort to delineate the relation between smoking and pain, the present study tested the attentional mediation model of smoking in pain perception. The authors hypothesized that by narrowing attentional focus, smoking in the presence of a distractor would facilitate distraction from painful stimuli, producing the most pronounced antinociceptive effect, compared with smoking or distraction alone. On the basis of previous findings of gender differences in pain, the authors hypothesized that the effects would vary by gender. The authors used the cold pressor test to assess pain threshold and tolerance in 76 smokers. A 2 (smoking/not smoking) x 2 (distraction/no distraction) x 2 (male/female) between-subjects analysis of variance (ANOVA) yielded Distraction x Gender interactions for pain threshold and pain tolerance. Pain sensitivity was lower in the distraction condition for men only. Whereas the results did not support the attentional mediation model of smoking in pain perception, they highlight the importance of distraction and gender differences in experimental pain. PMID- 15816317 TI - [Cardiac failure and life expextancy]. PMID- 15816318 TI - [Prevalence of Brugada syndrome among 35,309 inhabitants of Lorraine screened at a preventive medicine centre]. AB - Brugada syndrome is a recently identified cause of sudden death. Its primary prevention remains controversial, and epidemiology poorly defined. PATIENT POPULATION AND METHODS: Electrocardiograms (ECG) of 35,309 individuals (mean age = 37.2 years, 47% men) recorded over a 1-year period were reviewed and classified as (1) typical, (2) suspicious, and (3) negative. Subjects whose ECG was suspicious were offered a provocative test with flecainide, 2 mg/kg, i.v., and individuals whose ECG was typical were advised to undergo programmed ventricular stimulation (PVS). RESULTS: In 14 men and 6 women between the ages of 24 and 77 years (mean =47.5), ECGs were typical (n=6) or suspicious (n=14). Among 6 subjects with typical ECGs, 3 underwent PVS, which was positive in 1, who received an implantable cardioverter defibrillator (ICD). Among 14 subjects whose ECGs were suspicious, 5 declined further investigations and 5 developed typical ECG characteristics of Brugada syndrome after flecainide administration. PVS was negative in 4 subjects who consented to the procedure. Overall, among 35,309 individuals screened, 11 had ECG findings consistent with Brugada syndrome and, over a follow-up of 30 months, all had remained free of adverse cardiac event. CONCLUSIONS: we estimated a prevalence of Brugada syndrome of 0.3% in Lorraine. A single patient received an ICD for inducible ventricular tachyarrhythmia during PVS, representing a potential 30 per million asymptomatic adult rate of ICD implantation for this indication. PMID- 15816319 TI - [Biochemical detection of Conn's adenoma: definition of criteria and reference values]. AB - Primary hyperaldosteronism is a diagnosis which should be considered in refractory hypertension even in the absence of any hypokalaemia. Its detection relies above all on the levels of renin and aldosterone. The aldosterone/renin ratio has been proposed as the most sensitive criterium. The reference values used for the diagnosis of primary hyperaldosteronism are very variable in the literature, depending not only on the method used but also on the criteria used for their determination. In this study we evaluated the defined reference values prospectively by studying a population of patients with a Conn's adenoma treated surgically. The study included an initial retrospective period which allowed identification of 29 cases of Conn's adenoma treated surgically, and a 9 month prospective period during which 212 reports were collected. During this prospective period a further 9 cases of Conn's adenoma were detected, which were successfully treated with surgery. Analysis to discriminate the 38 Conn's adenomata from the rest showed that 3 parameters contributed significantly and independently to the diagnosis: supine plasma renin activity (ARPc), supine aldosteronaemia and the erect aldosterone/renin ratio, allowing correct classification in 88% of the cases. The reference ranges of these 3 parameters were calculated in order to give a sensitivity of 100% and the best possible specificity, therefore allowing a combined criterium involving all 3 parameters to be defined: ARPc < 0.45 ng/ml/h, supine aldosteronaemia >417 pmol/l, and erect aldosterone/renin >1180. PMID- 15816320 TI - [End-stage heart failure: quantity or quality of life?]. AB - Several clinical trials testing the safety and efficacy of different positive inotropic agents in patients suffering form end-stage heart failure have confirmed that these drugs increase mortality. We nevertheless, use these agents regularly because of the presumption that they improve quality of life. However, this presumption, sound a priori, is not firmly anchored. When we prescribe these drugs, do we inform our patients? Do the patients ultimately decide whether to be treated with inotropic agents? What is the role of palliative care in the end stage heart failure patient? We review the literature and reflect on these issues, long completely ignored, though beginning to elicit specific articles. PMID- 15816321 TI - [Sleep apnoea syndrome and cardiac failure]. AB - Despite recent therapeutic advances, chronic cardiac failure is still associated with a significant morbidity and mortality. Sleep apnoea syndrome is common in this population, affecting almost half of these patients. However, it is rarely diagnosed and treated. There are two types of sleep apnoea syndrome, which can sometimes co-exist: the obstructive apnoea syndrome with collapse of the upper airways, and the central apnoea syndrome with cyclical Cheyne-Stokes respiration, linked with anomalies of central control. Apnoea leads to sympathetic stimulation and an increase in the left ventricular post-charge which can alter cardiac function and the prognosis. Diagnosis of sleep apnoea syndromes is now made with small ambulatory oxymeters which do not disturb sleep and which allow precise detection of episodes of desaturation. Treatment with positive pressure ventilation brings an improvement in daytime symptoms (fatigue, drowsiness) as well as an improvement in cardiac function. Screening for sleep apnoea is thus essential in patients with chronic heart failure, especially in those resistant to optimal drug treatment, in order to improve their management. PMID- 15816322 TI - [A long-term follow-up quality evaluation of patients taking oral anticoagulant therapy]. AB - OBJECTIVES: the aim of this study was to estimate the population under Vitamin-K antagonists (VKA) in the region of Limousin (France), and to assess the treatment quality and the level of knowledge of the patients. METHODS: a transversal study permitted to identify all the patients under VKA under the French general health care coverage. During 6 months, a cohort of 370 patients had been followed. Major data measured were: the number of INR, the duration within the therapeutic range, the duration in the hazardous zone (INR >4.5), the level of patients' education and the knowledge on the treatment's indication by biologists. Data were collected through the French general health care database as well as questionnaires sent to general practitioners, biologists and patients. RESULTS: one percent of the entire population was under VKA. The mean number of INR performed during 6 months was 6.6, and 36.5% of patients had less than one biological control monthly. The mean duration passed within the therapeutic range was at 54%. The hazardous zone was reached by 22% of the patients. The level of education of the patients was poor in one half of the patients. The indication of the treatment was known in 6% of patients' biologists. CONCLUSION: these results reflect the limits of application of recommendations. Facing to this evidence, it is necessary to optimize the management and follow-up of these treatments, especially through the development of public health charters with biologists, education with patients, or even anticoagulant clinics. PMID- 15816323 TI - [Evaluation of the prevalence and management of normocholesterolaemic and non coronary hypertensives in France]. AB - We performed a study amongst hypertensive patients who consulted French general practitioners in the Sentinelles network. Firstly, the significance of this population in mainland France was quantified by keeping a register of all hypertensive patients aged over 40 years seen in a general practice consultation, and secondly the distribution of risk factors and management of normocholesterolaemic (total cholesterol < or = 2.5 g/l) without lipid lowering treatment and non coronary patients was described more precisely. Analysis of the population of 7864 hypertensives from this register showed that on average one doctor saw 3 hypertensive patients a day and that about half of them (54%) were normocholesterolaemic, non coronary patients. Analysis of this sub-population of 2519 patients showed that 19.9% of them had at least 3 of the other cardiovascular risk factors described in the ASCOT study (Anglo-Scandinavian Cardiac Outcomes Trial) and that 39.5% had controlled hypertension according to the New York Heart Association classification. The extrapolated number of hypertensive, non coronary patients who were normocholesterolaemic without lipid lowering treatment was estimated as 4.6 million (4.1:5.1) for the whole of France. PMID- 15816324 TI - [New energy sources for ablative methods]. AB - Radiofrequency current is the reference energy source for endocavitary ablation of arrhythmias. It is particularly well adapted for the ablation of focal arrhythmogenic substrates such as accessory pathways or foyers of automatism. Technological advances have made the lesions larger but the extension of the indications of percutaneous ablation to more complex substrates such as atrial fibrillation have justified the evaluation of alternative energies. The production of linear transmural lesions or deeper lesions which respect the parietal myocardial architecture and endocardial structure are a challenge for these energies. The capacity of functional mapping specific to cryogenics has provided this energy source with a clinical application for ablation of high risk structures whereas other energies, despite the chronicity of their experimental evaluation, are still at the stage of preliminary clinical trials with the sophisticated catheters in special indications. PMID- 15816325 TI - [The inter-individual variability of the response to clopidogrel]. AB - Clopidogrel is an anti-platelet aggregation drug with proven efficacy in the prevention of atherothrombotic complications. However, according to studies, between 5 and 20% of patients have further thrombotic episodes despite treatment with clopidogrel. The principal ex vivo evaluation methods of clopidogrel's efficacy are platelet aggregometry, and using flow cytometry to measure platelet activation and the quantatitive analysis of VASP phosphorylation. VASP analysis is the most selective method for clopidogrel's effect. These tests show great inter-individual variability in the response to treatment. This variability is such that about a third of treated patients show an "insufficient" response to clopidogrel. The exact causes of this variability have not yet been clearly identified. They could correspond with observer error, inter-individual variability in intestinal absorption or hepatic metabolism, or polymorphism of clopidogrel's target, the P2Y12 receptor. Accordingly it may be necessary to adapt the dose of clopidogrel or to use an alternative treatment. Failure to respond to clopidogrel would be a risk factor for atherothrombotic complications, but this association has yet to be demonstrated. PMID- 15816326 TI - [Microparticles and cardiovascular disease]. AB - Microparticles are membrane fragments liberated by activated or apoptopic cells. Thought for a long time to be cellular debris with no specific biologic function, in the vascular compartment they are a circulating reservoir of cellular effectors involved in thrombosis, inflammation, vascular remodelling and angiogenesis. High concentrations of circulating procoagulating microparticles found in many cardiovascular diseases indicate the importance of platelet, endothelial and monocytic activation and could contribute to the persistence of atherothrombotic disease. Pharmacological modulation of circulating microparticle concentrations could become a major therapeutic target in the future. PMID- 15816327 TI - The control of cardiomyocyte apoptosis via the beta-adrenergic signaling pathways. AB - Cardiomyocyte death resulting from apoptosis has been implicated in the evolution of heart failure. In this review, we focus on the concept that the cardiotoxicity of excessive sympathetic nervous system activity observed in heart failure is in part due to myocytes death by apoptosis. In vitro, high doses of norepinephrine induce adult cardiomyocyte apoptosis via 3-adrenergic receptor-coupled signaling pathways (PKA and Ca2+ entry-dependent mechanisms). beta1-and beta2-AR co-exist in the cardiac cell. beta1-AR stimulation is pro-apoptotic, whereas beta2-AR stimulation is anti-apoptotic, mediating its protective effect via coupling to Gi. These in vitro observations have been confirmed in transgenic mice: cardiac beta1-AR overexpression increases apoptosis and leads to heart failure, whereas cardiac beta2-AR overexpression has no deleterious effects. beta-AR stimulation activates p38 kinases and JNK (via the small GTP protein Rac1); and exert anti- and pro-apoptotic effects, respectively. Other studies suggest that beta1-AR stimulated apoptosis is dependent on Ca2+ -activated calmodulin kinase II and that the anti-apoptotic effect of beta2-AR is mediated via Akt-coupled pathways. beta-AR-stimulated apoptosis involves the mitochondrial pathway. Inhibition of mitochondrial permeability transition pore opening or caspase activation decreases beta-AR-stimulated apoptosis. Reactive oxygen species production is also involved in this process since superoxide dismutase/catalase-mimetics or catalase overexpression prevent beta-AR-stimulated apoptosis. In vivo, it has been shown that beta-AR blockers such as metoprolol and carvedilol have beneficial effects in animal models of chronic heart failure, associated with reduced apoptosis and improved cardiac systolic function. Understanding the mechanisms involved in the control of myocyte loss by the beta-adrenergic system will have direct clinical implications by improving the treatment of heart failure. PMID- 15816328 TI - Nitric oxide "at heart": emerging paradigms after a decade. AB - Despite the apparent redundancy of NOS isoforms in the myocardium, subcellular compartmentation dictates specific NO signaling from each isoform to colocalized effectors in response to physical (e.g. stretch) or receptor-mediated stimuli. Genetic deletion or overexpression experiments helped to characterize each isoform's respective role in the normal or diseased heart. eNOS and nNOS both contribute to sustain normal EC coupling and contribute to the early and late phases of the Frank-Starling mechanism of the heart. They also negatively modulate the beta1-/beta2-adrenergic increase in inotropy and chronotropy, and reinforce the (pre- and post-synaptic) vagal control of cardiac contraction, thereby protecting the heart against excessive stimulation by catecholamines. In the ischemic and failing myocardium, iNOS expression is induced and further contributes to attenuate the inotropic effect of catecholamines, as does eNOS coupled to overexpressed beta3-adrenoceptors. nNOS expression also increases in the aging and ischemic heart, but its role (compensatory or deleterious) remains to be defined. Many drugs currently used for the treatment of ischemic or failing cardiac diseases also activate and/or upregulate eNOS in the myocardium, which supports its proposed protective role, e.g. as "endogenous beta-blocker". Future pharmacologic modulation of the cardiac NOS will have to take into account their specific modulation of the various aspects of cardiac function, if one hopes to deliver more targeted and efficient therapy than currently achieved with exogenous NO donors. PMID- 15816329 TI - Rho proteins and vascular diseases. AB - As the cellular and molecular mechanisms of major arterial diseases such as atherosclerosis and hypertension are being more clearly defined, it is becoming apparent that these pathological processes share a number of functional and biochemical features in the vessel wall. Typically, arterial diseases are associated with functional and structural wall alterations including modified contractile properties, smooth muscle cell hypertrophy and proliferation, endothelial dysfunction, excessive extracellular matrix accumulation and inflammation. Small G proteins of the Rho family are defined as major regulators of cell functions including migration, proliferation, differentiation and gene transcription. Recent studies have demonstrated that activation of Rho proteins appears to be a common component for the pathogenesis of hypertension and vascular proliferative disorders. Functional analyses have further revealed that RhoA-dependent pathways are involved in excessive contraction, migration and proliferation associated with arterial diseases. This review focuses on the role of Rho proteins, in particular RhoA, in vascular smooth muscle cells and the involvement of Rho-dependent signaling pathways in vascular diseases. PMID- 15816330 TI - [Evaluation after 20 years of a case of Takayasu's disease that presented with aortic regurgitation]. AB - Takayasu's disease is a segmental multifocal affection of medium and large arteries. The diagnosis is based on the association of stenotic and aneurismal lesions of the aorta and its branches secondary to an inflammatory infiltration of the media and adventitia. Cases of aortic regurgitation associated with aneurismal dilatation of the ascending aorta as the presenting features of Takayasu's disease, as in this case, are rare. Histological examination of the aortic wall may help establish the diagnosis by showing signs of aortitis. The other usual arterial lesions are sometimes missing at the initial phase of the disease. A late histological diagnosis may be difficult as the inflammatory lesions tend to be progressively replaced by fibrotic lesions or a banal atheroma. PMID- 15816331 TI - [1/1 nodo-ventricular conduction atrial flutter with amiodarone]. AB - 1/1 atrial flutter is a regularly described complication of class I anti arrhythmics. It is, however, very rarely encountered with class III anti arrhythmics because prolongation of the atrio-ventricular node refractory period prevents 1/1 nodo-ventricular conduction. There have only been seven cases of 1/1 atrial flutter with amiodarone reported in the literature. Here we describe a new case of 1/1 atrial flutter with amiodarone. Our case clearly illustrates not only the different pro-arrhythmic effects of amiodarone (prolongation of the flutter cycle, and infra-Hissian block) but also the pathophysiological mechanisms possible with 1/1 conduction (prolongation of the flutter cycle, considerable permeability of the AV node). It demonstrates the difficulties of diagnosing such a rhythm disturbance, and that it is sometimes poorly tolerated, as well as underlining the importance of early diagnosis (in this case by oesophageal recording). Preventive treatment of 1/1 flutter can include amiodarone, digitalis, a betablocker or a bradycardic calcium inhibitor. PMID- 15816332 TI - [Treatment of acute myocardial infarction due to embolism by a thromboaspiration catheter]. AB - Although rare, coronary embolism poses a therapeutic problem. Thromboaspiration is an elegant alternative to implantation of a stent. The place of stenting in acute myocardial infarction depends on the results of randomised trials. The authors report the case of a 69 year old woman who presented with an apico lateral infarction probably due to a coronary embolism after postoperative thrombosis of a mechanical mitral valve prosthesis. The authors performed a percutaneous revascularisation by thromboaspiration with a good result. PMID- 15816333 TI - [Public health and medicine]. PMID- 15816334 TI - [Hearing screening of newborns. Preliminary results]. AB - Since 1976, an hearing screening is organized at the Clinic Edith Cavell. The testing was based on a behavioral technique of Veit-Bizaguet and, since 2001, the otoacoustic emissions are realised. The National Institutes of Health (1993) recommends universal newborn hearing screening of all newborns before 3 months of age and identification and treatment before 6 months of age. Indeed, detection of hearing loss and early intervention allow a better access to the language and consequently an easier schooling and social integration. This article shows the results of the screening from february at the end of December 2002. These data are compared with those of the literature. PMID- 15816335 TI - [Genetic predisposition and susceptibility to mycobacterial infections: the IL-12 IFNgamma axis deficiency]. AB - Since ten years severe pediatric infections which were idiopathic have now molecular explanation, defined new primary immunodeficiencies. These children presented a new kind of hereditary immunodeficiency with severe and/or recurrent infections caused by only one micro-organisms family. Some of these children have mycobacterial infections. Standard immunologic explorations were normal. However, they presented a vulnerability, sometimes lethal caused by Mycobacteria. The goal of this article is to describe the IL-12-IFNgamma axis deficiency (mendelian susceptibility to mycobacterial disease). PMID- 15816336 TI - [A new area for radiotherapy with favourable features]. AB - Radiotherapy is a complex medical speciality involving technology research, biology research and clinical research. All these basic researches are performed in order to optimise the management of cancer treatment patients. The aim of the present review is to present radiotherapy as a moving speciality whatever the concerned section. It will be particularly described the new approaches in terms of technology but also clinical developments. PMID- 15816337 TI - [Treatment of central precocious puberty with Gonadotropin-Releasing Hormone agonist (triptorelin) in girls: breast development, skeletal maturation, height and weight evolution during and after treatment]. AB - Central precocious puberty (CPP) is treated with GnRH analogues to stabilize secondary sexual characteristics and to prevent loss of final height (FH) due to accelerated bone maturation. However, some studies suggest that FH is not always improved and that treatment may induce excessive weight gain. We analysed data from 19 girls treated for CPP with monthly injections of 3.75 mg triptorelin. Pubertal development, bone age, height, weight and body mass index (BMI) were evaluated at start (chronological age: 7.8 +/- 1.8 yrs, mean +/- SD), at the end of treatment (10.6 +/- 1.1 yrs) and at FH (14.9 +/- 2.5 yrs). At start of treatment, breast (B) development was B3 (from 2 to 4), bone age 10.6 +/- 1.7 yrs, height 2.1 +/- 1.1 SDS and BMI 1.3 +/- 0.8 SDS. Treatment stabilized or reduced breast development and decreased bone maturation. Final height was 162.3 +/- 6.6 cm (0.0 +/- 1.1 SDS) and was comparable to predicted adult height at the start of treatment and to corrected mid-parental height. BMI SDS at the start, the end of treatment and at final evaluation were 1.3 +/- 0.8, 1.6 +/- 0.8 and 1.4 +/- 0.9 SDS. In conclusion, in our girls with central precocious puberty, treatment with GnRH agonist stabilized or decreased breast development and stabilized bone maturation, but did not increase neither final height nor weight. Aspects other than height should also be taken into account when considering treatment of children with precocious puberty. PMID- 15816338 TI - [Obesity in adult patients: check up and treatment]. AB - Obesity is now one of the major health problems in industrial countries as well as in developing world. Excess caloric intake and reduction of the physical activity are the main causes of obesity. This epidemic precedes a tremendous increase of type 2 diabetes, which is generally linked to weight excess. Obesity and type 2 diabetes are associated with morbidity and mortality and are very expensive for the social security. The important point is to define the risks linked to obesity taking into account the Body Mass Index and the importance of visceral obesity evaluated by waist measurement. After medical check up, a strategy will be discussed with the patient, including moderate caloric restriction and increased physical activity. Our patients and also some doctors suggest "popular diets" whose efficacy has not been demonstrated as superior. On a short time basis, low carbohydrate and high protein diets have some advantages, which can help our obese subjects but on long term, only hypocaloric and equilibrated diets are advisable. Drugs that proved their efficacy and tolerance may be prescribed in case of failure. Three drugs are presented, orlistat, sibutramine and metformine: their efficacy, secondary effects, interactions and finally their positioning. Bariatric surgery will be proposed to highly selected patients presenting morbid obesity. PMID- 15816339 TI - [Microscopic colitis]. AB - A 38-year old woman is admitted in the gastroenterology unit for a disabling episode of watery diarrhoea, not bloody and refractory to anti-diarrheic drugs. Different diagnostic exams remain negatives, with the exception of colic biopsies who disclose a lymphocytic colitis, one of the forms of the microscopic colitis entity. Microscopic colitis is an anatomo-clinic syndrome characterized by the presence of histological abnormalities on colic biopsies amongst patients suffering from chronic watery diarrhoea without endoscopic anomalies. Two clinical entities are today well known: collagenous colitis is characterized by a thickening of the sub-epithelial collagen band of the colon; lymphocytic colitis is defined as an increased level of lymphocytic cells, more than 20%, in the epithelial surface of colorectal mucosa. Inflammatory lesions of the chorion and alterations of the epithelial surface are seen in the two types of colitis. Their etiology remains unknown. It could be due to inflammatory lesions from autoimmune origin, activated by various exogenous agents such as bacteria and drugs. Clinical (feminine prevalence, frequent association with auto-immune diseases) and morphological similarities lead to argue and to hypothesis that both entities could represent different stages of the same disease, the lymphocytic colitis being the early stage. Various treatments have been proposed, for instance 5 aminosalicylates, but today mainly synthetic corticoids are used, especially budesonide. Others entities are recently been described: the chronic pericrypt eosinophilic enterocolitis and the colonic epithelial lymphocytosis ("epidemic"). PMID- 15816340 TI - [Chronic obstructive pulmonary disease (COPD): diagnostic aspects and therapeutic approaches]. PMID- 15816341 TI - [Felicien Rops: medicine, doctors, and his diseases (part one)]. AB - Felicien Rops was among the 19th-century's finest draughtsmen and Belgium's most sulphurous artist. Rops was born in Namur in 1833. He began to study law at the Free University of Brussels in 1851, joined a rowdy circle of students and began drawing the kind of caricatures and sharp satirical lithographs that would later gain him open-armed acceptance in the artistic and literary circles in Paris. As draughtsman and engraver, Rops has illustrated medical subjects such as "The massage", "Speculum", "Experimental medicine" "My gout". Rops was also a prolific letter-writer. In his correspondence, he spoke about his real or imaginary diseases: measles, gout, typhoid fever and typhus, congestion of the lung, phosphatous diabetes, nervous break-down, congestion of the brain, ocular injury, cardiac insufficiency. Rops' illnesses are discussed according to the medical knowledge at the time of Rops and to those of nowadays. PMID- 15816342 TI - Intergroup relations in soccer finals: people's forecasts of the duration of emotional reactions of in-group and out-group soccer fans. AB - The authors examined the hypothesis that people forecast a longer duration of uniquely human secondary emotions for their in-group than for an out-group. The authors conducted a field experiment in the setting of the European soccer championship. They asked Belgian participants to forecast the intensity with which their in-group Belgian fans or the out-group Turkish fans would experience various primary and secondary emotions in response to their team's victory or loss immediately after the Turkey-Belgium match and three days later. The results support the hypothesis. Moreover, and as the authors expected, they found no differences in the participants' forecasts of primary emotions. The authors discussed the implications of these findings for intergroup relations in general and for soccer fans' behavior in particular. PMID- 15816343 TI - Self-disclosure in romantic relationships and friendships among American and Japanese college students. AB - The author examined whether the level of self-disclosure would differ across four types of relationships--passionate love relationships, companionate love relationships, same-sex friendships, and cross-sex friendships--and across cultures: American culture and Japanese culture. Participants were 145 college students (64 Americans and 81 Japanese). The results supported three hypotheses: (a) Japanese students scored lower in self-disclosure than American students, regardless of relationship types, (b) self-disclosure was higher in same-sex friendships than in cross-sex friendships both among American participants and among Japanese participants, and (c) self-disclosure was higher in romantic relationships than in friendships both among American students and among Japanese students. However, the correlation between self-disclosure and passionate love was not stronger than the correlation between self-disclosure and companionate love. The author discussed the present study's findings and contribution. PMID- 15816344 TI - Family functioning, cultural orientation, and psychological well-being among university students in Lebanon. AB - The author examined the relationship between perceived family functioning, the cultural orientations of horizontal and vertical individualism and collectivism, and psychological well-being in a group of 182 university students in Lebanon. Factor analysis of the present data from the 12-item General Functioning Scale of the Family Assessment Device (GF-FAD) resulted in the extraction of one general factor, accounting for 43% of the variance. On the basis of North American cutoff values, the author found that 60.5% of the university students perceived their family as healthy and that 39.5% perceived their family as unhealthy. Male students perceived their families as somewhat more dysfunctional than did female students. Muslim students perceived their families as more dysfunctional than did Christian students. As the author expected, family functioning was associated with the culture-related personality dimension of horizontal collectivism and with perceived psychological well-being. Overall, the findings supported the structure and usefulness of the GF-FAD in the Lebanese context and the importance of viewing family functioning from the perspective of individual cultural orientation. PMID- 15816345 TI - The prediction of above-average participation in volunteerism: a test of the theory of planned behavior and the volunteers functions inventory in older Australian adults. AB - In the present prospective study of 81 older volunteers from a nonprofit organization in Australia, the authors compared the predictive utility of I. Ajzen's (1988) theory of planned behavior with that of E. G. Clary and M. Snyder's (1991) functional approach to volunteering. The authors mailed questionnaires to 385 volunteers in two waves of data collection. The first wave measured theory-of-planned-behavior variables and functional-approach variables. The second wave measured self-reported volunteering behavior for the previous month. Regression analyses supported both the theory of planned behavior and the functional approach; the theory of planned behavior accounted for a significantly larger proportion of variance in above-average participation in self-reported volunteerism. The findings of the present study provided some support for both the theory of planned behavior and the functional approach as models of self reported volunteerism. PMID- 15816346 TI - Interaction with a high-versus low-competence influence source in inductive reasoning. AB - Literature on inductive reasoning shows that when testing hypotheses, people are biased toward the use of confirmatory strategies (P. C. Wason, 1960). In the present article, the authors presented 2 studies showing how people use confirmation and disconfirmation strategies during actual interaction in problem solving. Study 1 showed that participants were able to learn to use disconfirmation when confronted with a low-competence, nonthreatening partner. When the partner was high in competence (thereby threatening the participant's competence), participants used confirmation, even when the partner used disconfirmation. In Study 2, the authors aimed at generalizing the aforementioned results by exploring the hypothesis that disconfirmation stems from the possibility of diverging from norms. Participants who were confronted with the violation of a conversational norm used a high proportion of disconfirmation, whatever the source of influence. When there was no violation but there was a low competence partner, the proportion of disconfirmation was high; when there was no violation but there was a high-competence partner, the proportion of disconfirmation was low. The authors discussed the interpersonal functions of confirmation and disconfirmation. PMID- 15816347 TI - The confederate's and others' self-conversion: a neglected phenomenon. AB - The authors analyzed a neglected phenomenon: self-conversion. Rather than concentrating on the influence exerted by a confederate on a naive participant, the authors studied how the confederate is himself or herself influenced by the position that he or she defends at the request of the experimenter. This experiment is particularly revealing with respect to a fundamental aspect of the psychology of the proselytizing of minority groups: In the process of convincing others, one also convinces oneself. Exerting influence on others thus leads to self-conversion, which allows the group to reinforce its beliefs and to stand up to conformity pressures. PMID- 15816348 TI - A social-value analysis of postmaterialism. AB - The author investigated the relationship between social values and R. Inglehart's (1971, 1981, 1990) Postmaterialism concept. In his theory, Materialism and Postmaterialism are basic value dimensions motivated by a person's need for security and short-term survival (on the one hand) and the desire to address needs that transcend material concerns (on the other). The aim of the present study was to attempt to locate Postmaterialism relative to politically nonspecific social values. Participants were 161 New Zealand university undergraduate men and women who completed P. R. Abramson and R. Inglehart's (1995) Postmaterialism scale and a 56-item Social Values Inventory (S. H. Schwartz, 1992). Analyses of Postmaterialism scale scores and aggregated scores reflecting the motivations underlying different groups of social values supported the hypothesis that Postmaterialism, as measured by this scale, is positively related to values associated with Self-Direction and Universalism motivations and negatively related to Security motivations. The results indicate that previous research had been limited to the application of values representing only a small part of the overall values space. PMID- 15816349 TI - Audience perceptions of candidates' appropriateness as a function of nonverbal behaviors displayed during televised political debates. AB - Compared to televised debates using a single-screen format, such debates using a split screen presenting both debaters simultaneously show viewers the nonverbal reactions of each debater's opponent. The authors examined how appropriate or inappropriate such nonverbal behaviors are perceived to be. Students watched one of four versions of a televised debate. One version used a single-screen format, showing only the speaker, whereas the other three versions used a split-screen format in which the speaker's oppodent displayed constant, occasional, or no nonverbal disagreement with the speaker. Students then rated the debaters' appropriateness. Analysis indicated that the opponent was perceived to be less appropriate when he displayed any background disagreement compared to when he did not. The students perceived the speaker as most appropriate when his opponent displayed constant nonverbal disagreement. PMID- 15816350 TI - The effectiveness of compliance techniques: foot in the door versus door in the face. PMID- 15816351 TI - Experimental approaches to evaluate respiratory allergy in animal models. AB - Asthma is defined as a chronic disease of the entire lung and asthma attacks may either be immediate, delayed or dual in onset. Allergic asthma is a complex chronic inflammatory disease of the airways and its etiology is multifactorial. It involves the recruitment and activation of many inflammatory and structural cells, all of which release mediators that result in typical pathological changes of asthma. A wealth of clinical and experimental data suggests that allergic asthma is due to an aberrant lung immune response mediated through T-helper type 2 (Th2) cells and associated cytokine-signaling pathways. The pathology of asthma is associated with reversible narrowing of airways, associated with prominent features that involve structural changes in the airway walls and extracellular matrix remodeling including abnormalities of bronchial smooth muscle, eosinophilic inflammation of the bronchial wall, hyperplasia and hypertrophy of mucous glands. The primary objective of respiratory allergy tests is to determine whether a low-molecular-weight chemical (hapten) or high-molecular-weight compound (antigen) exhibits sensitizing properties to the respiratory tract. This may range from reactions occurring in the nose (allergic rhinitis), in the bronchial airways (i.e., allergic bronchitis, asthma) or alveoli (e.g., hypersensitivity pneumonitis). Current assays utilize several phases, viz. an induction phase, which includes multiple exposures to the test compound (sensitization) via the respiratory tract (e.g., by intranasal or intratracheal instillations), by inhalation exposures or by dermal contact, and a single or multiple challenge or elicitation phase. The challenge can either be with the chemical (hapten), the homologous protein conjugate of the hapten or the antigen. The choice depends both on the irritant potency and the physical form (vapor, aerosol) of the hapten. The appropriate selection of concentrations (dosages) both for the induction and elicitation of respiratory allergy appears to be paramount for the outcome of test. Endpoints to characterize positive response range from the induction of immunoglobulins, cytokine or lymphokine patterns in serum (or the lung) to (patho-)physiological reactions typifying asthma. None of the currently applied animal models duplicate all features of human asthma. Accordingly, the specific pros and cons of the selected animal model, including protocol variables, animal species and strain selected, must be interpreted cautiously in order to arrive at a meaningful extrapolation for humans. PMID- 15816352 TI - Gene expression profiling in streptozotocin treated mouse liver using DNA microarray. AB - Streptozotocin (SZ) is known to exert toxic effects not only on pancreatic islet beta cells but also on other organs including liver. For analyzing changes in genes expression associated with SZ toxicity, we performed DNA microarray analyses on the liver obtained from SZ-treated mice. Eight-week-old male ICR mice were treated i.p. with 200 mg/kg of SZ, and the blood and liver were taken at 6, 24 and 48 h after the treatment. Labeled cRNA prepared from total RNA of the liver was hybridized to the GeneChip Murine Genome U74A V.2 (Affymetrix). The number of the probe sets, which were clearly up-regulated or down-regulated, were over 100 at 6 and 24h after the SZ-treatment, and it decreased at 48 h after the treatment. Many of the up-regulated genes were categorized into cell cycle/apoptosis related genes, immune/allergy related genes and stress response/xenobiotic metabolism related genes. On the other hand, genes related to glucose, lipid and protein metabolisms were down-regulated. These changes started prior to the elevation of the serum glucose levels, indicating the direct action of SZ on the liver rather than the secondary effect of diabetes. This may be related with the previously reported hepatic changes such as lipid peroxidation, mitochondrial swelling and inhibition of hepatocyte proliferation observed before the development of hyperglycemia. PMID- 15816353 TI - Morphological and gene expression analysis in mouse primary cultured hepatocytes exposed to streptozotocin. AB - Streptozotocin (SZ) is known to exert toxic effects not only on pancreatic islet beta cells but also on other organs including the liver. For analyzing direct effects of SZ on hepatocytes, we performed morphological analysis and DNA microarray analysis on mouse primary cultured hepatocytes. Hepatocytes were taken from non-treated Crj:CD-1(ICR) mice. The primary cultured hepatocytes were treated with SZ at concentrations of 0, 1, 3, 10, 30 and 100 mM. After the treatment for about 6 or 24h, cell survival assay using tetrazolium salt (WST-1), light microscopic/electron microscopic analysis and gene expression analysis were performed. For the gene expression analysis, target (labeled cRNA) prepared from total RNA of the hepatocytes was hybridized to the GeneChip Murine Genome U74A V.2 (Affymetrix). The signal intensity calculation and scaling were performed using Microarray Suite Software Ver 5.0. IC50 of the cell survival assay was around 62 mM at 6 h exposure and 7 mM at 24 h exposure. Marked chromatin margination was observed in nuclei of the hepatocytes treated with SZ at concentrations of 3 or 10mM. Gene expression analysis revealed similar expression changes to those of in vivo, i.e. up-regulation in cell proliferation/ apoptosis related genes, and down-regulation of lipid metabolism related genes. These results potently supported the hypothesis that many of the hepatic alteration including histopathological and gene expression changes are induced by direct effect of SZ rather than by the secondary effect of the hyperglycemia or hypoinsulinemia. PMID- 15816354 TI - Neonatal tamoxifen treatment of mice leads to adenomyosis but not uterine cancer. AB - Tamoxifen is contraindicated during pregnancy but many births have been reported in breast cancer patients taking this drug and numbers might be expected to increase with FDA approval of tamoxifen for risk reduction in women at high, risk of breast cancer. The neonatal mouse, exquisitely sensitive to xenobiotic estrogens, has been used to investigate the effects of short-term oral dosing with tamoxifen (1 mg/kg on days 2-5 after birth) on long-term changes in uterine pathology and gene expression. Increased adenomyosis incidence and severity was evident in the tamoxifen-treated mice with increasing age. Uterine weights in treated mice remained lower than the corresponding controls up until 9 months, after which they became greater but during life-time studies (up to 36 months), there was no development of uterine tumours. Pathological examination of uterine tissues showed there to be extensive down-growth of endometrial glands and stroma into thickened, abnormal myometrium that had disorganised fascicles of smooth muscle and increased interstitial collagen deposition. In advanced cases, the endometrial epithelium showed mild degrees of focal hyperplasia and squamous metaplasia but no atypical cytology suggestive of premalignant change. Microarray analysis of uterine RNA taken at 1.5, 3, 6, 9 and 12 months showed from 4500 ESTs, only 12 genes were continuously over-expressed by tamoxifen treatment over this time, while none was continuously down-regulated. Up-regulated genes include those for nerve growth factor (Ngfa), cathepsin B (Ctsb), transforming growth factor beta induced (Tqfbi) and collagens (Colla1, Colla2). Results provide a basis for understanding the mechanism for tamoxifen induced tissue remodelling and the development of adenomyosis. PMID- 15816355 TI - Decrease of the surface fraction of surfactant proteins containing clara cells and type II pneumocytes in a rat asthma model. AB - In asthma surfactant proteins (SP) might differ in distribution and composition and thus play a role in pathophysiology of this disease. Therefore, the well established animal model of ovalbumin sensitized and challenged rats were used to study the distribution of surfactant proteins in Clara cells and type II pneumocytes. Serial sections of paraffin embedded lung tissue were sequentially immunostained by the avidin-biotin-peroxidase complex (ABC) technique. Antisera against SP-A, SP-B and Clara cell specific protein (CC10) were used. We determined stereologically' the surface fraction of immunolabelled cells and semiquantitatively the percentage of test fields containing labelled alveolar macrophages. In allergen sensitized and provocated rat lungs: (1) the surface fraction of SP-A and SP-B positive Clara cells was significantly reduced, (2) the surface fraction of Clara cells stained with CC10 was coincided with controls, (3) the surface fraction of SP-A and not of SP-B possitive type II pneumocytes decreased significantly, (4) a significantly higher percentage of test fields with SP-A labelled alveolar macrophages was evaluated. Thus, in this animal model of asthma the inflammatory process after allergen challenge is accompanied by alterations in the distribution patterns of SP in Clara cells and type II pneumocytes. PMID- 15816356 TI - Role of mast cells in hepatic remodeling during cholestasis and its resolution: relevance to regulation of apoptosis. AB - BACKGROUND/AIMS: Mast cells are thought to be related to fibrogenesis, but recent studies have shown that fibrosis of the liver can be induced even in mast cell deficient rats. To clarify the significance of mast cell accumulation in cholestatic liver diseases, the relations between such accumulation, bile ductule proliferation and apoptosis of biliary epithelial cells were examined in the rats during cholestasis and its resolution. METHODS: Cholestasis and its resolution were induced in rats by common bile duct ligation and spontaneous recanalization, respectively. The extent of bile ductule proliferation and the numbers of mast cells and apoptotic biliary epithelial cells were estimated quantitatively in liver sections. RESULTS: Recanalization of the ligated common bile duct led to an abrupt and transient increase in the number of mast cells, although the number of proliferated bile ductules decreased rapidly. The number of apoptotic biliary epithelial cells of the proliferated bile ductules increased rapidly and transiently, and the change paralleled that of the mast cells. CONCLUSIONS: Mast cells accumulating in the portal triads during cholestasis and its resolution may relate to the reduction of proliferated bile ductules, i.e., in hepatic remodeling, through the induction of apoptosis of biliary epithelial cells. PMID- 15816357 TI - Role of adhesion molecule ICAM in the pathogenesis of polymicrobial sepsis. AB - INTRODUCTION: Intercellular adhesion molecule-1 (ICAM-1) is thought to be involved in polymorphonuclear leukocytes (PMNL) recruitment and secondary organ damage in response to infection and inflammation. The precise role of ICAM-1 in disease progression is unknown and remains a topic of controversy. The aim of this study was to investigate the effect of ICAM-1 on histological changes and cytokine synthesis in a murine model of polymicrobial sepsis. METHODS: Polymicrobial sepsis was induced in experimental animals by caecal ligation and puncture (CLP). A control group was formed using sham laparotomy without CLP. In order to ascertain the role of ICAM-1 in the response, procedures were performed in both ICAM-1 knockout animals (ICAM-1-/-) and in C57BL/6 mice that were not genetically modified (wild type, WT). Clinical response was observed daily, morphological changes occurring in the lung and liver were studied using light microscopy and quantified using a scoring system. Plasma concentrations of various cytokines (TNF-alpha, IL-6, IL-10) were measured via ELISA. RESULTS: In ICAM-1-/- mice a less severe clinical response to induced sepsis was observed with significantly less weight loss and hypothermia. A significantly lower mortality rate was observed in ICAM-1-/- mice (12.5% vs. WT: 45.5%) and no significant histological changes were apparent in pulmonary or hepatic tissue on light microscopy following CLP. In WT animals however, significant evidence of leukocyte infiltration and interstitial thickening in pulmonary tissue was observed. Similarly, hepatic tissue sinusoidal widening and hydropic degeneration was present. In addition, pro- and anti-inflammatory cytokine synthesis in ICAM-1 /- animals was significantly attenuated when compared to WT mice. (ICAM-1-/-: TNF alpha: 67.7+/-12.1pg/microl; IL-6: 208.9+/-26.7pg/microl; IL-10: 34.6+/ 5.8pg/microl; WT: TNF-alpha: 840.7+/-150.2pg/microl; IL-6: 3100.2+/-1052.3 pg/microl; IL-10: 1550.1+/-495.7 pg/microl). DISCUSSION: This study suggests that ICAM-1 has an important pathophysiological role in the response to polymicrobial sepsis. It would appear that absence of this molecule impairs the ability of PMNL to migrate into organ tissues and reduces consequent secondary organ damage resulting in improved clinical status and overall survival. Further investigation into the effectiveness of ICAM-1 modulation in the treatment of sepsis is warranted. PMID- 15816358 TI - PGE2 alleviates kidney and liver damage, decreases plasma renin activity and acute phase response in cirrhotic rats with acute liver damage. AB - In this study, we evaluated the effect of prostaglandin E2 (PGE2) on renal and hepatic function using an experimental cirrhosis model plus acute liver damage (ALD). Male Wistar rats treated with carbon tetrachloride (CCl4) for 8 weeks were used for the cirrhosis model. Cirrhotic rats were further exposed to an additional acute dose of CCl4 to induce ALD and then treated with PGE2 intramuscularly twice a day for 7 days (200 microg/Kg/day). PGE2 administration started 3 h after the additional dosing of CCl4 and PGE2 effect on hepatorenal function was examined on days 1, 2, 3, and 7. PGE2-treatment ameliorated the decrease in urinary sodium excretion, and normalized serum activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and plasma renin observed in cirrhotic rats with ALD. In addition, PGE2-treatment decreased mean arterial pressure, glomerular hypercellularity and thickening of the kidney capillary wall, and liver steatosis and cellular necrosis. Also, PGE2 increased the number of regenerative nodules. Finally, PGE2-treatment inhibited the increase in Alpha 1-acid glycoprotein (pAGP), fibrinogen, and Apo A-1 mRNA expression by 83%, 59%, and 77%, respectively. These results suggest that PGE2 administration may decrease the expression of acute phase proteins. In conclusion, PGE2-treatment improved hepatic and renal function and may be useful to down-regulate the acute phase response in cirrhotic rats presenting ALD induced by CCl4. PMID- 15816359 TI - Selected biochemical parameters and ultrastructural picture of pancreas due to Ulinastatin treatment of experimental acute pancreatitis. AB - Urinary trypsin inhibitor (UTI, Ulinastatin) is a protease inhibitor that has not been yet used in Europe in any experimental trial of severe acute pancreatitis. We have combined the experimental model of severe, hemorrhagic form of acute pancreatitis, and pharmacological treatment with a protease inhibitor. Male Wistar rats were divided into four experimental groups: healthy controls, operated, operated with experimentally induced acute pancreatitis, and animals with acute pancreatitis--treated with UTI preparations. Subjects in the last group were administered UTI intraperitoneally 1 h after pancreatitis induction in an average standard dose of 3000 units/animal. Additionally, four subgroups were created in this treated group, based on the UTI administration time--number of standard doses received: 2 h - 1 standard dose, 6 h - 5 standard doses, 12 h - 11 doses, 24 and 48 h - 15 doses. Statistically significant differences in the serum amylase and lipase activity between the UTI-treated and non-treated subjects were found. In the group of non-treated animals, there a profound destruction of cellular organelles was observed with a total degradation of nuceli, endoplasmatic reticulum and zymogen granules. However, in the UTI-treated subjects, pathological processes proceeded with the significantly slower pace and in much smaller quantities. PMID- 15816360 TI - Local and systemic toxicity of h-R3, an anti-epidermal growth factor receptor monoclonal antibody, labeled with 188osmiun after the intracerebral administration in rats. AB - h-R3 is a humanized anti-epidermal growth factor receptor (EGFR) monoclonal antibody (MAb). This receptor is over-expressed in the majority of tumors of epithelial origin, including glioblastomas. 188Rhenium (188Re) constitutes an ideal radionuclide for imagining and radioimmunotherapy, and its toxicity is known, nevertheless, it is unknown if 188Os, as 188Re's daughter, has any local or systemic toxicity effect when it is administered intracerebrally for treating intracranial tumors. For this reason we decided to assess the toxicity of stable 188Os once the complete decay of 188Re has occurred, by administering intracerebrally to rats the h-R3 labeled with 188Os. Forty rats (20 each sex) were distributed randomly into four experimental groups (ten per group): control group received 5microL of glucoheptonate solution vehicle; two other groups were treated with unlabeled or labeled h-R3 with 188Os. The remaining group served as a non-treated control group. A single 5 microL dose (2.5 microL into each lateral ventricle) of neutral solution containing 50 microg of h-R3 labeled initially with 13.25 microCi of 188Re was stereotactically administered into lateral ventricles 8 days after the conjugation with the radionuclide was done. Each animal was observed daily for detection of toxicity signs. Body weights were recorded on days 0, 7 and 14. Blood samples for analysis of hematological and clinical chemistry parameters were taken on days 0 and 14. Necropsy and histopathological studies were carried out at the end of the study. All animals gained weight by day 14. There were no changes in hematological and clinical chemistry, but minimal histopathological changes were observed at the application sites. This study shows that single doses of 188Os-h-R3 is tolerable and causes minimal local and no systemic toxicity effects in rats. PMID- 15816361 TI - Effects of pioglitazone on hyperglycemia-induced alterations in antioxidative system in tissues of alloxan-treated diabetic animals. AB - Hyperglycemia not only generates reactive oxygen species but also attenuates antioxidant mechanisms creating a state of oxidative stress. Oxidative stress is thought to play a crucial role in pathogenesis of chronic diabetic complications. Pioglitazone is a new oral antidiabetic agent, a potent inhibitor of glycation and potent antioxidant. In the present study, normoglycemic and alloxan-induced diabetic rabbits were treated with pioglitazone (1 mg/kg daily) for 4 and 8 weeks. At the end, glutathione peroxidase (GSH-Px), glutathione reductase (GSSG R), glutathione (GSH) and protein carbonyl groups (PCG) were evaluated in homogenates of liver and kidney. Chronic hyperglycemia caused a marked increase in oxidative processes and some changes in activity of antioxidants. In liver, diabetic vs. control values (mean+/-S.E.M; P<0.05) for GSH-Px were 181.0+/-5.4 vs. 203.1+/-1.9 and 187.4+/-6.6 vs. 240.9+/-18.8mU/mg protein. Pioglitazone treatment for 8 weeks affected GSH-Px activity in diabetic liver (261.5+/-7.3 mU/mg protein). In diabetic kidney, GSSG-R activity (20.6+/-1.6 vs. 32.4+/-1.5 and 23.6+/-0.6 vs. 36.3+/-0.3 mU/mg protein) and GSH level (16.6+/-0.5 vs. 23.2+/ 0.9 and 17.9+/-0.5 vs. 23.2+/-0.6 nmol/mg protein) were diminished, while PCG level (0.32+/-0.03 vs. 0.11+/-0.02 and 0.35+/-0.03 vs. 0.16+/-0.03 nmol/mg protein) was elevated. In diabetic kidney, pioglitazone restored to control values GSSG-R activity (34.4+/-1.4 and 30.6+/-0.1 mU/mg protein) as well as GSH (25.5+/-1.2 and 21.6+/-0.5 nmol/mg protein) and PCG (0.16+/-0.01 and 0.19+/-0.02 nmol/mg protein) levels. The present study showed that pioglitazone reduced to some extent the oxidative stress enhanced by chronic hyperglycemia. PMID- 15816362 TI - Proglumide attenuates experimental colitis in rats. AB - Ulcerative colitis is associated with altered contractile activity and transit time of colon. On the other hand, cholecystokinin (CCK) has been shown to play an important role in regulation of gastrointestinal motor function including colonic contraction and transit. In the present study, an attempt was made to study the effect of proglumide, a CCK receptor antagonist on experimental colitis in rats. Experimental colitis was induced in male Sprague-Dawley rats by instilling 1 ml of 4% acetic acid followed by flushing with 0.5 ml air. The rats were kept in a head-down position for 30s. Finally, each rat received 1.5 ml colonic wash with 1.5 ml saline. Four groups of rats received proglumide orally (0, 250, 500 and 1000mg/kg). The first dose of proglumide was given 1 h before acetic acid challenge, whereas the second dose of proglumide was given 25 h after the first dose. Sham control rats received an equal volume of saline instead of acetic acid. Forty-eight hours after the acetic acid challenge, the colon was removed, weighed and split longitudinally and scored for injury. Part of the colon was used for histopathological study as well as analysis of myeloperoxidase (MPO) activity (as a marker of neutrophil activity). Acetic acid produced severe diarrhea and exfoliation of the colonic epithelium accompanied by extensive destruction of the mucosal interstitium. Proglumide dose dependently protected rats against acetic acid-induced increase in colon weight, diarrhea, MPO activity and colonic injury. Inhibition of CCK exerts a beneficial effect in experimental colitis. Further studies are warranted to determine the mechanism of protection and the therapeutic potential of CCK inhibitors. PMID- 15816363 TI - Postural instability of extremely obese individuals improves after a body weight reduction program entailing specific balance training. AB - The purposes of this study were to compare postural stability between obese and lean subjects and to investigate the effect of a 3-week body weight reduction (BWR) program entailing specific balance training on postural stability of extremely obese patients. Time of balance maintenance and mean error on the medial-lateral direction at the trunk and lower limb level were assessed during a single limb stance on a movable platform in 19 non-obese and in 20 extremely obese individuals (age range: 20-40 yr). Time of balance maintenance was shorter (obese: 21.1+/-7.7 vs lean: 27.3+/-3.1 sec) and medial-lateral sway of the trunk was larger in obese (5.4+/-3.2 degrees) than in lean (3.2+/-1.1 degrees) subjects (p<0.05). Two subgroups of obese subjects were also tested after a BWR program (energy-restricted diet, moderate physical exercise, nutritional education and psychological counselling) combined with or without 6 sessions of specific balance training on a movable platform. BWR plus specific balance training enhanced time of balance maintenance (pre: 23.8+/-7.2 vs post: 30.0+/-0.0 sec) and reduced the trunk sway (5.2+/-2.8 degrees vs 2.6+/-0.9 degrees ) more than BWR alone (p<0.05). The present findings indicate that extremely obese individuals have inadequate postural stability (compared to their lean counterparts) that could however be improved by few sessions of specific balance training incorporated into a multidisciplinary BWR program. It was concluded that balance improvement is an important goal of rehabilitation, that would probably reduce the propensity of overweight individuals to fall while performing everyday activities. PMID- 15816364 TI - B-type natriuretic peptide in patients with clinical hyperthyroidism. AB - AIMS: To investigate the level and clinical significance of plasma B-type natriuretic peptide (BNP) in patients with hyperthyroidism. METHODS: Plasma BNP was measured in 32 healthy subjects and 67 patients with clinical hyperthyroidism. Left ventricular anatomy and function was assessed with echocardiography. RESULTS: The average BNP in hyperthyroid patients was higher than that in healthy subjects (186+/-140 vs 34+/-17 ng/l, p =0.001). The increase in BNP was mainly found in hyperthyroid patients who had clinical and echocardiographic evidence of left ventricular dysfunction (250+/-190 ng/l). The BNP level in patients with hyperthyroidism, but with normal left ventricular function, was similar to that of the controls (37+/-17 vs 34+/-17, ng/l, p>0.05). Multi-regression analysis showed that left atrial diameter and left ventricular ejection fraction were independently associated with the plasma levels of BNP. CONCLUSIONS: there is a significant elevation in plasma BNP in patients with hyperthyroidism; the increase is largely due to hyperthyroidism-induced left ventricular dysfunction. Measurements of plasma BNP may help to detect heart failure in patients with clinical hyperthyroidism. PMID- 15816365 TI - Prevalence of osteopenia in men with prolactinoma. AB - The aim of this cross-sectional study was to analyze bone mineral density (BMD) and prevalence of osteopenia and osteoporosis in 30 men with prolactinoma, and compare them to 22 control subjects. BMD of lumbar spine and femur was evaluated by dual-energy X-ray absorptiometry. PRL, testosterone, estradiol, sexual hormone binding globulin and free androgen and estrogen indexes (FAI and FEI, respectively) were measured in all the subjects. In patients with prolactinoma, mean values of PRL and testosterone were calculated for the 12-month period that preceded the study. The mean T-score of the four sites analyzed by bone densitometry was lower in men with prolactinoma than in controls (p-values: lumbar spine=0.015, femoral neck <0.0001, trochanter=0.037, total femur=0.036), and 55.6% of the former presented osteopenia or osteoporosis at one or more sites (p =0.035). The lumbar spine was the most seriously affected site, where 29.6% had osteopenia and 14.8% had osteoporosis. By the time of BMD determination, significant associations were found between BMD and PRL, testosterone, FAI, estradiol, FEI, and duration of hypogonadism. Considering the period of 12 months that preceded BMD evaluation, trochanter BMD was associated with mean PRL levels, while there was an association between lumbar spine BMD and mean testosterone levels. However, the multiple regression analysis showed that estradiol was the main determinant of BMD. In conclusion, men with prolactinoma have high prevalence of osteopenia and osteoporosis. Bone loss in such patients is associated with hyperprolactinemia and hypogonadism, and mainly influenced by estrogen. PMID- 15816366 TI - Experience in management of 51 non-functioning pituitary adenomas: indications for post-operative radiotherapy. AB - OBJECT: The indications for additional radiotherapy (RT) after surgery for non functioning pituitary adenoma are controversial. The goal of this retrospective study was to evaluate the outcome of surgically treated patients, with or without post- operative irradiation. METHODS: Review of cases treated for non-functioning pituitary adenoma. Fifty-one patients were identified, with a mean post-operative follow-up of 6.4+/-3.5 yr. Twenty-nine patients showed residual tumor after surgery and 22 did not. Serial endocrine, visual and radiological evaluations were made after treatment to assess the efficacy and toxicity of surgery and RT. Twenty-seven patients with residual tumor after surgery received RT (22 of them during the post-operative period and 5 after an interval of several yr: 3 because of increased tumor size and 2 with stable residual lesion); tumors in 14 of these patients decreased in size, 11 appeared to be stable on imaging and one patient showed some increase in tumor size (one patient was not followed-up). The residual tumors of the 2 non-irradiated patients remained stable after 5 and 7 yr, respectively. Twenty-two patients without residual disease after surgery (11 with post-operative irradiation, 1 with RT 5 yr after transsphenoidal surgery because of tumor recurrence, and 10 without RT) have shown no evidence of tumor regrowth on serial images. CONCLUSIONS: Radiotherapy can be avoided in patients with complete macroscopic resection and absence of residual tumor in post operative images; they must be carefully followed using imaging techniques and, in the case of recurrence, they should be re-operated and/or irradiated. PMID- 15816367 TI - Reappraisal of the risk of iodine-induced hyperthyroidism: an epidemiological population survey. AB - The occurrence of iodine-induced hyperthyroidism (IIH) has been reported after iodine supplementation from clinics and hospitals, but not following an epidemiologic survey. We studied the prevalence of thyroid derangement in a population following iodine supplementation. One yr after more than 75% of the population had been consuming 40 ppm iodized salt; information regarding history of endemic goiter and iodized salt production, distribution, consumption and monitoring were collected in four cities of the Islamic Republic of Iran. A total of 6048 subjects were randomly selected. All subjects were assessed for size of goiter, and urinary iodine and serum T4, T3, TSH, anti-thyroglobulin and anti thyroperoxide were measured. Before iodine supplementation, all four cities were areas of endemic goiter. The rate of household consumption of iodized salt was 50, 75 and 90% in 1994, 1995 and 1996, respectively. Ninety-one percent of the salt samples contained 15-55 ppm iodide. Total goiter rate was 57, 62 and 68%; median urinary iodine was 188, 197 and 190 microg/l in the age groups of 6-18, 19 40 and >40 yr, respectively. Prevalence of clinical and subclinical hyperthyroidism was 0.34 and 0.41 and those of clinical and subclinical hypothyroidism were 0.51 and 1.07%, respectively. Nine point eight and 18% in the 19-40 yr age group and 17.6 and 25.6% in >40 yr old subjects had positive anti thyroperoxidase and anti-thyroglobulin, respectively. This systemic epidemiologic study in an iodine deficient population showed that, following a well-executed iodine supplementation program, the occurrence of IIH is rare. PMID- 15816368 TI - Mutational analysis of the inhibin alpha gene in preeclamptic women. AB - BACKGROUND: Preeclampsia (PE) is a disorder that occurs in at least 5% of pregnancies and affects both the mother and the unborn baby. A dramatic increase of maternal serum inhibin A concentration in the second and third trimester of pregnancy is a common feature of PE and inhibin A measurement may add significant prognostic information for predicting PE in pregnant women. DESIGN: We evaluated the presence and prevalence of gene polymorphisms for inhibin alpha subunit (INHalpha) in patients affected by PE (no.=50; study group), and in the general population (control group composed of 103 women and 42 men). METHODS: DNA extraction, single strand conformation polymorphism analysis, DNA sequencing, restriction fragment length polymorphism analysis, and Fisher's exact test were used. RESULTS: A 769G-->A transition was found in INHalpha1, but not in INHalpha2 or INHalpha3 fragment. This variant was found in 10/145 normal controls (7,6%), and in 1/50 preeclamptic patients (2%), without significant difference between the two groups (p=0.29). CONCLUSIONS: The prevalence of INHalpha gene variants is not increased in PE. Due to its frequency, the 769G-->A transition may be considered a polymorphism present in the general Italian population. PMID- 15816369 TI - Inflammatory markers in women with a recent history of gestational diabetes mellitus. AB - Gestational diabetes mellitus (GDM) is a risk factor for both Type 2 diabetes (DM2) and insulin-resistance syndrome (IRS). C-reactive protein (CRP), fibrinogen and leukocyte count are increased in the IRS and predict DM2 and cardiovascular disease (CVD). The chemochine monocyte chemoattractant protein-1 (MCP-1/CCL2) is also elevated in DM2 and CVD. Recent evidence suggests a relation between chronic inflammation and GDM, but post-delivery information on inflammatory markers in these high-risk women is lacking. Serum levels of CRP, fibrinogen, MCP-1/ CCL2, and leukocyte blood count have been assessed in 26 women with and 26 women without a recent history of GDM, matched for age, body mass index (BMI), post partum duration and parity. DM2 was excluded in all the participants by an oral glucose tolerance test (OGTT). Women with previous GDM showed significantly higher CRP (p=0.007) and fibrinogen (p=0.02) serum concentrations, whereas MCP 1/CCL2 serum levels and leukocyte blood count were comparable in the two groups. Overall, CRP levels significantly correlated with BMI (r=0.40, p=0.03), waist-to hip ratio (WHR) (r=0.44, p=0.001), fasting insulin (r=0.27, p=0.04), insulin resistance assessed by means of the homeostatic model (HOMA) (r=0.28, p=0.04), and fibrinogen concentration (r=0.49, p=0.0001). At linear regression analysis, only WHR and fibrinogen were independently associated with CRP levels. In conclusion, the increase of inflammatory markers may be one of the first detectable disorders in healthy women at high risk of DM2 and IRS, like those with a GDM history. PMID- 15816370 TI - Hyperprolactinemia affects spermiogenesis in adult male rats. AB - The mechanisms underlying the antifertility effects of hyperprolactinemia have yet to be established in an appropriate experimental model. Hyperprolactinemia is a known side effect of fluphenazine, a broad spectrum, long-acting phenothiazine known to be dopamine type-D2 receptor antagonist. In our earlier study in adult male rats, we reported that fluphenazine at a dose of 3 mg/kg/day suppressed serum FSH but not testosterone (T) through increasing dopamine (DA) metabolism in the pituitary gland, within 60 days. Fluphenazine treatment affected sperm quality and male rats treated with fluphenazine sired fewer litters. The effects of fluphenazine-induced hyperprolactinemia on sperm quality appeared to be related to reduced FSH. We now report that FSH suppression enhanced the uptake of acridine orange (AO), a DNA intercalating, fluorescent dye by the fluphenazine treated caput epididymal sperms with concomitant reduction in the uptake of thiol specific monobromobimane (mBBr) fluorescent dye in vitro, suggesting greater accessibility of DNA intercalating dye to sperm chromatin and reduction in free sperm protein thiols. The concomitant increase in AO and decrease in mBBr fluorescence was suggestive of loose chromatin packaging in caput epididymal sperms after treatment with fluphenazine at 3 mg/kg/day for 60 days. The suppression in levels of protamine (P1) in caput epididymal sperms suggested that chromatin hypocompaction was due to reduced deposition of protamines in sperm chromatin. Reduction in testicular levels of cyclic adenosyl 3', 5' monophosphate response element modulator (CREMtau) and P1 further suggested that reduced deposition was indeed due to reduced synthesis. The concomitant reduction in testicular levels of transition protein 1 (TP1) and transition protein 2 (TP2) also suggested that hypoprotamination was due to reduced synthesis of these proteins crucial for facilitating P1 deposition. The effect appeared to have occurred at the level of translation of CREMtau, since its transcript levels were unaffected whereas those of TP1, TP2 and P1 and protamine were upregulated. The study led to the view that the effects of FSH suppression were manifest on the posttranscriptional modifications of CREMtau, as also on transcript repression of TP1, TP2, P1, which do the RNA- binding proteins bring about. Reduction in FSH did not decrease ABP expression in the testis, which has recently been implicated in the expression of transition protein 1 in vitro. However, a significant reduction was evident after fluphenazine treatment, in the immunoexpression of testicular cAMP, the mediator of FSH effects in the Sertoli cells and putative mediator of ABP effects in the spermatids. The study suggests that fluphenazine induced hyperprolactinemia suppressed FSH and affected a putative cAMP-dependent mechanism underlying posttranscriptional modification of spermatidal genes involved in chromatin condensation, presumably by reducing the availability/secretion of ABP, a paracrine regulator of spermiogenesis in vitro. PMID- 15816371 TI - Spironolactone in the treatment of polycystic ovary syndrome: effects on clinical features, insulin sensitivity and lipid profile. AB - This prospective clinical trial was designed to assess the effects of a long-term therapy with spironolactone, with and without dietary-induced weight-loss, on clinical features, lipid profile and insulin levels in women with polycystic ovary syndrome (PCOS). Twenty-five patients (range of age 16-32 yr; 13 lean and 12 overweight) fulfilling formal diagnostic criteria for PCOS (oligomenorrhea and/or amenorrhea, biochemical and/or clinical evidence of hyperadrogenism) were studied at baseline and then received oral spironolactone (100 mg/die) for 12 months; association with lifestyle modifications was recommended to all over weight patients. Clinical, endocrine and metabolic parameters [oral glucose tolerance test (OGTT), lipid profile] were measured at baseline and at the end of the antiandrogen treatment. The therapy was associated with a significant average decline of triglycerides in overweight subjects and with increased HDL cholesterol levels in lean patients. The insulin levels at 60 min during OGTT, homeostasis model assessment-insulin resistance and area under curve of insulin were significantly lowered in overweight women after 12 months of spironolactone and weight loss and no negative changes in insulin secretion and sensitivity were observed in PCOS women after pharmacological treatment alone. The efficacy of spironolactone on the androgenic clinical aspects of PCOS has been confirmed in this study. Furthermore, our data show that long-term treatment with spironolactone exerts no negative effects on lipoprotein profile and glucose metabolism; more relevant beneficial effects on glucose and lipid metabolism were observed when the antiandrogen was associated with weight loss in overweight PCOS women. PMID- 15816372 TI - A six month mitotane course induced sustained correction of hypercortisolism in a young woman with PPNAD and Carney complex. AB - A low-dose mitotane (MT) regimen was evaluated as a pharmacological approach for correcting the severe hypercortisolism in a young woman affected by Carney complex (CNC) and primary pigmented nodular adrenocortical disease (PPNAD). In the first 12 week period, the MT daily dose was progressively increased from 0.5 to 4.0 g/day. This dosage was maintained for an additional 16 weeks (cumulative dose 602 g, plasma MT maximum level 12 microg/ml), and then stopped because of sustained signs of hypoadrenalism requiring prednisone replacement. Complete regression of seborrhea, acne, and plethora was observed after 8 weeks of treatment (cumulative dose 95 g). Regular menses returned after 13 weeks (cumulative dose 197 g, plasma MT 8 microg/ml). Profound decrease of both serum cortisol (from 615 to 220 nmol/l) and urinary free cortisol (UFC) values (from 1498 to 477 nmol/day) was noted after 16 weeks of treatment (cumulative dose 314 g, plasma MT 8 microg/ml). MT treatment was associated with mild gastric discomfort and reversible increase of cholesterol plasma levels. Low serum cortisol and UFC were still observed 41 weeks after MT was discontinued (plasma MT 0.2 microg/ml). Our report demonstrates that low dose MT treatment may be a safe and effective modality for a sustained correction of hypercortisolism by PPNAD in subjects with CNC waiting for surgery. PMID- 15816373 TI - Adrenocortical carcinoma producing 11-deoxycorticosterone: a rare cause of mineralocorticoid hypertension. AB - A 37-yr-old man presented with the classic signs of mineralocorticoid excess hypertension and hypokalemia. The cause was not aldosterone excess, but elevation of plasma 11-deoxycorticosterone (DOC). Computed tomography (CT) scans showed a large right adrenal mass without signs of metastatic disease. The tumor was removed by open laparotomy, and histology revealed an adrenocortical carcinoma. Two yr after diagnosis, the patient is in good general condition and there is no sign of recurrence or metastatic disease, despite the large tumor size. DOC producing adrenocortical carcinomas causing mineralocorticoid hypertension are very rare, so far only 10 cases have been described in the literature. PMID- 15816374 TI - Thyroid hemiagenesis and incidentally discovered papillary thyroid cancer: case report and review of the literature. AB - Thyroid hemiagenesis (TH) is a rare congenital abnormality in which one thyroid lobe fails to develop. Its prevalence is uncertain, because the absence of one thyroid lobe does not usually cause clinical symptoms. The detection of TH is usually incidental when the evaluation of other thyroid disorders is requested. It is more frequently found in female than in male patients (3:1 ratio) and in the left lobe compared to the right lobe. We report the case of a 54-yr-old man, presenting with a large multinodular right-sided goiter, with mediastinal extension and dysphagia. Thyroid scan and ultrasound study showed the absence of the left lobe. The patient underwent surgery for compressive symptoms, and the operation confirmed the absence of the left lobe. Histological examination demonstrated a multi-nodular goiter with papillary carcinoma. To our knowledge, this case represents the first reported case of association between TH and papillary thyroid carcinoma in a male patient, and the second in which the tumor arose in the right lobe. PMID- 15816375 TI - Use of radioguided surgery with [111In]-pentetreotide in the management of an ACTH-secreting bronchial carcinoid causing ectopic Cushing's syndrome. AB - Intraoperative [111In]-pentetreotide scintigraphy with a hand-held gamma detector probe has recently been proposed to increase the intraoperative detection rate of small neuroendocrine tumors and their metastases. We report a case of a 28-yr-old woman with ectopic Cushing's syndrome due to an ACTH-secreting bronchial carcinoid, in whom the use of radioguided surgery improved disease management. At presentation, radiolabeled pentetreotide scintigraphy was the only procedure able to detect the ectopic source of ACTH. After radiologic confirmation, the patient underwent removal of a bronchial carcinoid, with disease persistence. After surgery, pentetreotide scintigraphy showed pathologic uptake in the mediastinum not previously detected at surgery and only subsequently confirmed by radiologic studies. Despite a second thoracic exploration, hormonal, scintigraphic, and radiological evidence of residual disease persisted. Radioguided surgery was then performed using a hand-held gamma probe 48 h after iv administration of a tracer dose of radiolabeled [111In-DTPA-D-Phe1]-pentetreotide, which permitted detection and removal of multiple residual mediastinal lymph node metastases. Clinical and radiologic cure, with no evidence of tracer uptake at pentetreotide scintigraphy, was subsequently observed. The use of an intraoperative gamma counter appears a promising procedure in the management of metastatic ACTH-secreting bronchial carcinoids. PMID- 15816379 TI - [Urologic cancer. Prevention and screening]. PMID- 15816378 TI - Three dancing dwarves. PMID- 15816380 TI - [Epidemiology of kidney tumors]. PMID- 15816376 TI - The role of luteinizing hormone activity in controlled ovarian stimulation. AB - The role of LH in the natural menstrual cycle is undisputed. The active participation of LH in both steroidogenesis and ovulation is well established, but its potential effect on oocyte maturation in the issue of assisted reproduction protocols remains a topic of debate. Although several studies have added to our understanding of the specific actions of androgens in human follicular development, some discrepancies persist regarding their role in oocyte atresia. Clinical situations, where LH is either decreased or absent (e.g. in women with hypogonadotrophic hypogonadism or LH-receptor gene mutations), provide important data supporting the necessity for a minimal amount of LH to evoke ovulation. Recent use of GnRH antagonists, which results in profound suppression of LH concentration, in combination with the pharmacological production of recombinant gonadotrophins, has attracted the attention of investigators. Identification of sub-fertilized women, in whom LH administration could be beneficial and should be indicated, is arousing ever more interest. Based on the available data in the literature, the aims of this review are to assess the role of both endogenous and exogenous LH activity in stimulated cycles, and to evaluate the effects of recombinant human LH supplementation on the ovarian hormonal milieu and on the main outcomes of controlled stimulated cycles. PMID- 15816381 TI - [Renal cell carcinoma and environmental factors]. PMID- 15816382 TI - [Role of arterial hypertension in kidney cancer]. PMID- 15816383 TI - [Native kidney tumors and kidney failure]. PMID- 15816377 TI - Endocrine disturbances in depression. AB - Depression is one of the most common psychiatric disorders. For a long time, clinicians suspected a causal link between depression and the endocrine system. The most frequently occurring endocrine abnormality in depressed subjects is hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis. CRH and AVP are likely to play a substantial role in the pathophysiology of this disorder, and their receptors appear to be a specific target for future antidepressant drugs. Depression also affects the hypothalamic-pituitary-GH (HPGH) and -thyroid (HPT) axes. Alterations in the reproductive system may also play a role in the pathology of depression. In addition, there is increasing evidence that leptin and neurosteroids, such as DHEA, are implicated in mood disorders. PMID- 15816384 TI - [Hereditary forms of kidney cancer and genetic screening]. PMID- 15816385 TI - [Epidemiology of bladder cancer]. PMID- 15816386 TI - [Risk factors of bladder tumors excluding occupational risks]. PMID- 15816387 TI - [Occupational bladder tumors: practical modalities for their identification in occupational disease]. PMID- 15816388 TI - [Which role and which tools in the screening of bladder cancer today?]. PMID- 15816389 TI - [Urothelial pseudoneoplastic and preneoplastic lesions of the bladder]. PMID- 15816390 TI - [Urothelial tumors induced by chemotherapy and radiotherapy]. PMID- 15816391 TI - [Genetic features of urothelial carcinoma of the bladder and the upper urinary tract]. PMID- 15816392 TI - [Epidemiology of penile cancer]. PMID- 15816394 TI - [Epidemiology of testicular cancer]. PMID- 15816393 TI - [Prevention and screening of penile cancer]. PMID- 15816395 TI - [Risk factors for testicular cancer and carcinogenesis mechanisms]. PMID- 15816396 TI - [Carcinoma in situ of the testis]. PMID- 15816397 TI - [Scrotal self-examination]. PMID- 15816398 TI - [Which role for scrotal ultrasonography?]. PMID- 15816399 TI - [Secondary tumors in patients whose testicular cancer has been cured]. PMID- 15816400 TI - [Genetics and germinal tumors of the testis]. PMID- 15816401 TI - [Epidemiology of prostatic cancer]. PMID- 15816402 TI - [Chemoprevention of prostatic cancer]. PMID- 15816403 TI - [Management in case of isolated precancerous lesions of the prostate (HG-PIN) discovered with biopsy]. PMID- 15816404 TI - [Prostate cancer screening]. PMID- 15816405 TI - [Androgen replacement therapy and prostate cancer]. PMID- 15816406 TI - [Genetic features of prostatic cancer]. PMID- 15816407 TI - Current progress in shrimp endocrinology-a review. AB - One problem in aquaculture is obtaining brood because many commercially important species are incapable of spontaneous maturation under artificial conditions. Commercial shrimp hatcheries commonly use eyestalk ablation to stimulate gonadal maturation in shrimps. Research has been conducted on the inhibition of reproductive maturation by hormones originating in the eyestalk glands and on other endocrine sources (e.g.,brain, thoracic ganglion, ovary, mandibular organ, androgenic gland and Y-organs) to determine their roles. Alternate techniques for acceleration of gonad maturation through the use of synthetic hormones or neurotransmitters may benefit aquaculture. Neurohormones and neuroregulators have been shown to accelerate gonadal maturation but an effective delivery technique must be developed for use in a large-scale aquaculture operation. PMID- 15816408 TI - Seasonal effects of melatonin on ovary and plasma gonadotropin and vitellogenin levels in intact and pinealectomized catfish, Clarias batrachus (Linn). AB - Effects of daily administration of melatonin for 15 days were evaluated with respect to ovarian activities and plasma gonadotropin (GtH II) and vitellogenin (Vg) levels in intact (INT) and pinealectomized (Px) female catfish, C. batrachus, during preparatory (April), prespawning (May and June), spawning (July) and post-spawning (September) periods. Px (saline control groups) caused a stimulatory effect during preparatory (with respect to Vg synthesis and incorporation) and prespawning (with respect to Vg synthesis) periods whereas no effect was observed during spawning and post-spawning periods with respect to the reproductive parameters studied. During April, melatonin-treatment significantly decreased plasma GtH II levels and percentage of vitellogenic oocytes without any significant changes in plasma Vg levels and gonadosomatic index (GSI). During early prespawning period, in May, 50microg melatonin brought about a significant reduction in plasma GtH II levels in INT group, whereas 100microg caused a decrease in all parameters; on the other hand, in Px groups both dose levels proved to be inhibitory. In June (late prespawning period) melatonin-treatment could not bring about any change in GSI and plasma Vg levels compared to the control groups regardless of Px but plasma GtH II and mean number of yolky oocytes were significantly reduced in melatonin-treated INT group. During spawning period (July) melatonin inhibited the GSI, mean number of yolky oocytes and plasma GtH II levels without affecting plasma Vg levels. In September (post spawning period), melatonin did inhibit both GSI and plasma GtH II levels. The results, thus, indicate that melatonin showed variable effects (inhibitory and/or no effect) to GSI, mean number of yolky oocytes and plasma Vg levels but a consistent inhibiton of plasma GtH II levels indicating that melatonin may control the reproduction by blocking the GtH II release from the pituitary via affecting the hypothalamo-hypophysial axis. PMID- 15816409 TI - Clobetasol propionate solid lipid nanoparticles cream for effective treatment of eczema: formulation and clinical implications. AB - In the present study clobetasol propionate (Cp) was loaded as solid lipid nanoparticles (SLN), incorporated it in suitable cream base and evaluated in vitro and its performance clinically against equivalent marketed formulation. Cp was incorporated into SLN by high-pressure homogenization technique and characterized for mean particle size, surface morphology and per cent drug entrapment. Drug permeation and skin uptake studies from Cp creams were carried out in a validated Franz static diffusion cell across human cadaver skin (HCS). Sixteen chronic eczema patients were enrolled in a controlled double blind clinical trial. Optimized Cp-SLN was smooth and spherical under scanning electron microscopy; with average particle size of 177 nm and per cent drug entrapment of 92.05%. In vitro permeation studies revealed lower mean flux value and higher skin uptake of Cp from Cp-SLN cream compared to marketed drug cream. Both formulations were found to be responsive to manifestations of chronic eczema, while Cp-SLN cream prepared in this investigation registered significant improvement in therapeutic response (1.9 fold; inflammation, 1.2 fold; itching) in terms of per cent reduction in degree of inflammation and itching against marketed cream. Further clinical trials are required to ascertain the efficiency of the present formulation. PMID- 15816410 TI - Effect of losartan and enalapril on cognitive deficit caused by Goldblatt induced hypertension. AB - The present study was designed to evaluate the learning and memory, in an altered physiological state associated with increased blood pressure and activated renin angiotensin system in Wistar rats. The role of angiotensin in cognitive function was assessed by treatment with angiotensin converting enzyme (ACE) inhibitor enalapril (2 mg/kg), angiotensin 1 receptor (AT(1)) antagonist losartan (5 mg/kg) and their combination. The experimental renal hypertension was induced by the method of Goldblatt. Learning and memory was assessed using the radial arm maze test. Acetylcholine esterase (AChE) levels in the pons medulla, hippocampus, striatum and frontal cortex were measured as a cholinergic marker of learning and memory. Results indicate that in comparison to normotensive rats, renal hypertensive rats committed significantly higher number of errors and took more trials and days to learn the radial arm maze learning and exhibited memory deficit in the radial arm maze retrieval after two weeks of retention interval, indicating impaired acquisition and memory. Treatment with enalapril, losartan and their combination attenuated the observed memory deficits indicating a possible role of renin angiotensin system in cognitive function. AChE level was reduced in hippocampus and frontal cortex of renal hypertensive rats which could be attributed to the observed memory deficit in hypertensive rats. It can be concluded that, renal hypertensive rats had a poor acquisition, retrieval of the learned behavior, perhaps a possible disturbance in memory consolidation process and that this state was reversed with ACE inhibitor enalapril and AT 1 receptor antagonist losartan. PMID- 15816411 TI - Effect of licofelone against NSAIDs-induced gastrointestinal ulceration and inflammation. AB - The present study was aimed to evaluate the effect of licofelone, a dual inhibitor of cycloxygenase1/2-5-lipoxygenase against indomethacin-induced gastric damage in rats and mice in order to assess the role of leukotrienes if any, in non-steroidal anti-inflammatory drugs (NSAIDs)-induced gastrointestinal inflammation. Acute pretreatment with licofelone reversed the indomethacin induced gastric ulceration, neutrophil adhesion in mesentery venules, neutrophil count in blood, lipid peroxides and vascularity in the stomachs of mice and rats. Further, chronic pretreatment of licofelone also prevented indomethacin-induced gastric morphological changes and cellular infiltration in mesentery venules. Moreover, acute administration of indomethacin elevated leukotriene B4 levels in gastric mucosa, which was reversed by pretreatment with licofelone The results suggest that licofelone offered gastroprotection against NSAIDs-induced gastropathy through its effect on leukotrienes and by inhibiting extravasation of neutrophils. PMID- 15816412 TI - In vivo effect of whole grain flour of finger millet (Eleusine coracana) and kodo millet (Paspalum scrobiculatum) on rat dermal wound healing. AB - Influence of finger millet and kodo millet on rat dermal wound healing was assessed by making a 4 cm2 (2 x 2 cm) excision wound on the shaven back of rats under ether anesthesia. Finger millet or kodo millet flour (300 mg) as aqueous paste was applied topically once daily for 16 days. The granulation tissue formed on day 4, 8 and 12 was used to estimate some biochemical parameters like protein, DNA, collagen and lipid peroxides. There was significant increase in protein and collagen contents and decrease in lipid peroxides. Biophysical parameters like rate of contraction and number of days for epithelialization were also studied. Rate of contraction was 88-90% in kodo millet and finger millet treated rats in comparison to 75% in untreated rats. The number of days for complete closure of wounds was lower for finger millet (13 days) and kodo millet (14 days) treated rats in comparison to untreated (16 days) rats. The results implicate a possible therapeutical role for finger millet and kodo millet in accelerating the process of wound healing. PMID- 15816413 TI - Influence of zinc on the status of hepatic trace elements and biokinetics of 65Zn in ethanol treated rats. AB - Whole body counting studies of 65Zn indicated that the Tb1 (the faster component) was significantly decreased while the slower component (Tb2) was increased significantly following ethanol treatment. Interestingly, following zinc treatment to ethanol treated rats, slower component (Tb2) of 65Zn came back to within normal limits while the faster component (Tb1) got significantly elevated in comparison to ethanol treatment. Percent uptake values of 65Zn were found to be increased in liver, intestine, muscle, brain and kidney, and decreased in bone under alcoholic conditions. Interestingly, the uptake values of 65Zn in all the organs except muscle were reverted back to within normal limits upon zinc supplementation to these ethanol intoxicated animals. A significant decrease in zinc contents was noticed in ethanol treated rats, which, however, were raised to normal levels upon zinc supplementation: Copper levels, on the other hand, were significantly enhanced in both ethanol fed and combined ethanol + zinc treated rats. Calcium levels were significantly decreased in both ethanol and zinc treated rats, which however were further reduced upon zinc supplementation to ethanol fed rats. However, no significant change was observed in the concentrations of sodium and potassium in any of the treatment groups. In conclusion, zinc appears to play a protective role by normalizing the turnover of 65Zn in whole body as well as in its uptake in different organs under alcoholic conditions. PMID- 15816414 TI - Efficacy of Terminalia arjuna (Roxb.) on N-nitrosodiethylamine induced hepatocellular carcinoma in rats. AB - The effect of ethanolic extract of Terminalia arjuna bark on carbohydrate metabolizing enzymes of N-nitrosodiethylamine induced hepatocellular carcinoma in Wistar albino rats were studied. The plasma and liver glycolytic enzymes such as hexokinase, phosphoglucoisomerase, aldolase were significantly increased in cancer induced animals while glyconeogenic enzyme, glucose-6-phosphatase was decreased. These enzymes were reverted significantly to near normal range in treated animals after oral administration of T. arjuna for 28 days. The modulation of the enzymes constitute the depletion of energy metabolism leads to inhibition of cancer growth. This inhibitory activity may be due to the anticancer activity of constituents present in the ethanolic extract of T. arjuna. PMID- 15816415 TI - Effect of glurenorm on immunohistochemical changes in pancreatic beta cells of rats in experimental diabetes. AB - Immunohistochemical localization of islets of Langerhans of streptozotocin (65 mg/kg, ip) induced diabetic + glurenorm (10 mg/kg, po) treated female albino rats revealed increase in number of beta cells and insulin immunoreactivity of beta cells. The results suggest that glurenorm can cause the stimulation of beta cells of endocrine pancreas in diabetic rats. PMID- 15816416 TI - Restriction analysis of conserved and variable regions of VP2 gene of Indian isolates of bluetongue virus serotype 1. AB - Bluetongue virus (BTV) is a member of Orbivirus genus in family Reoviridae. The virus genome is composed of 10 double-stranded RNA segments. The RNA segment L2 encodes an outer capsid viral protein VP2, which is the main determinant of neutralization and serotype-specific immune response. BTV serotype 1 (BTV-1) specific novel primer pair was designed using VP2 gene sequences available in GenBank to amplify 1240-1844 bp region because two hypervariable and three conserved regions have been reported within these 604 nucleotides. This primer pair successfully amplified cell culture adapted six Indian isolates of BTV-1 from different geographical regions of the country. The 604 bp PCR product of VP2 gene of all six BTV-1 yielded two fragments of 273 and 331 bp when digested with Taq1 restriction enzyme. This indicated that there is only one TaqI site at 1513 bp (within 1240-1844 bp region) of VP2 gene of BTV-1 Indian isolates. The in silico restriction analysis revealed that in BTV-1 South African isolate (BTV 1SA) there is no TaqI site while in BTV-1 Australian isolates (BTV-1AUS), there are two TaqI sites (at 1513 and 1567 bp) within 1240-1844 bp region of VP2 gene. The earlier reported VP2 gene based primer pair for BTV-1 was used in the present study to amplify 2242-2933 bp region of six BTV-1 Indian isolates as three conserved regions have been reported within these 691 nucleotides. The digestion of 691 bp PCR products with XmnI yielded three fragments of 364, 173 and 154 bp with all the six Indian isolates of BTV-1 suggesting that there are two XmnI sites within 2242-2933 bp region of VP2 gene. A single XmnI site was observed in silico in BTV-1AUS and BTV-1SA isolates at different positions within this region. The in vitro and in silico restriction profile analyses of partial VP2 gene sequences using TaqI and XmnI restriction enzymes indicated a close relationship of Indian isolates of BTV-1 with BTV-1AUS isolates but not with BTV 1SA isolate. PMID- 15816417 TI - A simple method for efficient extraction and purification of C-phycocyanin from Spirulina platensis Geitler. AB - Phycocyanin is a major light harvesting accessory pigment of red algae and cyanobacteria. In the light of its many commercial applications in food and pharmaceutical industry, purity of the pigment plays a major role. Pharmaceutical industry demands a highly pure phycocyanin with A620/280 ratio of 4 and food industry a ratio of 2. In the present study phycocyanin was extracted in sodium phosphate buffer (pH 7) after macerating in liquid nitrogen. The crude phycocyanin thus extracted was precipitated with 50% ammonium sulphate, purified by dialysis and finally by gel filtration chromatography. Pure phycocyanin was finally obtained with an A620/A280 value of 4.98. PMID- 15816418 TI - In vitro propagation of Dendrobium hybrids using flower stalk node explants. AB - Large-scale in vitro propagation protocol for Dendrobium hybrids Sonia 17 and 28, two highly prized commercial cut flower cultivars through shoot multiplication using flower stalk node explants and protocorm-like bodies (PLBs) formation was accomplished. Both hybrids did not exhibit significant differences in initiation, multiplication, rooting, and field establishment. Flower stalk nodes cultured on half strength Murashige and Skoog (MS) medium supplemented with 6.97 microM kinetin (Kn), or 15% coconut water (CW) or 13.3 microM of N6-benzyladenine (BA) evoked bud break. Kn showed better growth of the initiated bud. Excision and culture of the initiated shoots on medium having same amount of Kn developed more than 5 shoots per shoot directly from the base. Subsequent culture enhanced the rate of shoot induction. Transfer of isolated shoots onto 44.4 microM of BA enriched medium displayed induction of more than 6 PLBs from the base within 60 days. PLBs underwent rapid multiplication upon transferral to medium having the same concentration of BA (44.4 microM). Subsequent culture increased the proliferation of PLBs. No decline was observed in the proliferation of shoots as well as PLBs up to 15th subculture. PLBs transferred onto half strength MS medium with 6.97 microM of Kn underwent conversion of more than 90% PLBs to shoots. The shoots were rooted at the best on half strength MS medium with 2 g l(-1) activated charcoal. Survival rate of the plantlets of the two hybrid cultivars after acclimatization was more than 80%. PMID- 15816419 TI - Inter-specific hybridization between freshwater catfish Mystus cavasius (Ham & Buch) and M. seenghala (Sykes) by artificial fertilization. AB - By employing the technique of induced ovulation for artificial fertilization, inter-specific hybrids between the threatened catfish species (Mystus cavasius x M. seenghala) were produced. Fertilization, hatching and survival were significantly different between control and hybrids. The hatching time of the hybrid was significantly lower than that of the control. The average performance viz., hatching time and viability of larvae of the control fish, was significantly better than that of the hybrids. In the hybrid cross, hatchlings were mostly deformed and abnormal and after yolk absorption ultimately succumbed. PMID- 15816420 TI - In vivo induction of sister chromatid exchanges (SCEs) and chromosomal aberrations (CAs) by lynestrenol. AB - Genotoxicity study of synthetic progestin lynestrenol, was carried out on mouse bone marrow cells using sister chromatid exchanges (SCEs) and chromosomal aberrations (CAs) as parameters. Lynestrenol was studied at three different doses (6.87, 13.75 and 27.50 mg/kg body wt.). SCE and CA increased significantly as compared to normal control when treated with lynestrenol at 13.75 and 27.50 mg/kg body wt. The present results suggest that lynestrenol has both a genotoxic and cytotoxic effects in mouse bone marrow cells. PMID- 15816423 TI - The draft pharmaceutical policy: the weakest link. PMID- 15816421 TI - Effect of nifedipine and amlodipine on dead space wound healing in rats. AB - Effect of two calcium channel blockers (CCBs) nifedipine and amlodipine, was studied on normal and steroid depressed wound healing in albino rats, using the dead space wound model. The drugs enhanced normal healing as evidenced by increase in tensile strength of 10 days old granulation tissue. There was neither a significant change in the hydroxyproline level (or collagen) nor a change in the glycosaminoglycan content in granulation tissue. However, lysyloxidase level was increased significantly. The increase in tensile strength could thus be attributed to better cross-linking and maturation of collagen rather than collagen synthesis per se. The drugs were also able to overcome steroid depressed wound healing. It is likely that the prohealing effects may be related to the improved antioxidant status too, since superoxide dismutase levels were observed to be higher in the CCB- treated animals. PMID- 15816424 TI - [Pain indicators in critical care]. AB - In this descriptive/qualitative study, focus groups and individual interviews were completed with 48 critical care nurses and 12 critical care physicians. Results showed that many indicators are used by those clinicians for pain assessment in intubated patients, whatever their level of consciousness. Nurses and physicians mentioned giving weight to the patient's self-assessmen tof pain even when intubated, communication then being by means of signs. Behavioural and physiological indicators also provide clinicians with valuable information for pain assessment. However, for the most part these indicators are not specific to pain, limiting their usefulness in this context. The study aslo suggests recommendations for nursing practice and ideas for nursing resarch. PMID- 15816425 TI - [Campaign against smoking: how a few minutes could save lives]. PMID- 15816426 TI - [Animal bites and rabies]. PMID- 15816427 TI - [Ten good reasons to use the Need for Interagency Services]. PMID- 15816428 TI - Efficacy of transarterial chemoembolization prior to liver transplantation for hepatocellular carcinoma as found in pathology. AB - BACKGROUND/AIMS: To analyze the efficacy of chemoembolization prior to liver transplantation in liver explants. METHODOLOGY: We reviewed pathological findings in the explanted livers of 21 patients with histologically proven hepatocellular carcinoma and liver cirrhosis who underwent transarterial chemoembolization (TACE) prior to liver transplantation. Nine patients had solitary nodules with a median diameter of 4 cm (range 1.5-7 cm), 7 patients had 2 or 3 tumors with a median total diameter of 5.9 cm (range 3-9 cm) and 5 patients had a multifocal tumor prior to TACE. Pathological up-staging of the clinical tumor classification was documented as "tumor-progression." Concurrence of clinical and pathological findings was documented as "steady disease". "Tumor regression" described those cases in which the pathological classification downgraded the clinical findings. RESULTS: There was no treatment-related morbidity in these patients' group. Tumor regression was proved in 11/21 patients (52.4%) whereas steady disease was observed in 7/21 patients (33.4%). In 5 patients (23.8%) no vital tumor was found by pathological examination. Tumor regression was observed only in one of the five patients having a multifocal tumor prior to TACE. Tumor progression was observed in 3/21 patients (14.3%). CONCLUSIONS: Our data show that TACE provides acceptable local tumor control as bridging treatment before liver transplantation. Although the majority of our patients (15/21, 71.4%) had 2 or more tumor lesions at the beginning of treatment, tumor progression was observed in only a minority (14.3%) of patients. However, multifocal tumors could not be successfully under-staged through this treatment and, furthermore, vital tumor was always observed in pathology; the usefulness of TACE in multifocal disease has to be re-estimated. PMID- 15816429 TI - Anatomical and physiological comparison of liver volumes among three frequent types of parenchyma transection in live donor liver transplantation. AB - BACKGROUND/AIMS: Adequate venous outflow is a prerequisite for successful live donor liver transplantation. Several techniques of liver transection have been established in both transplant and non-transplant hepatic surgery. The purpose of our study was to define and compare anatomical and physiological characteristics of venous drainage in the three most common types of liver transection techniques (Malago, Cantlie, Pringle) encountered in live donor liver transplantation METHODOLOGY: Volumes of both graft and remnant livers as well as individual hepatic vein territories were calculated by means of virtual 3-dimensional reconstructions of computed tomography images obtained from 55 potential live liver donors using the software HepaVision (MeVis, Germany). Belonging to the middle hepatic vein (MHV) was assigned according to the largest territorial volume on either right or left hemilivers. Livers from all potential donors were subject to virtual splitting (n=55). Findings were subsequently confirmed intraoperatively in those who underwent donor resection (n=27). RESULTS: There were no statistically significant differences in hemiliver-volumes among the three types of liver partition. There was a predominance of MHV belonging to the right hemiliver: Malago n=45 (82%) vs. Cantlie n=44 (80%) vs. Pringle n=44 (80%). Dominant right MHV hemiterritory showed a mean volume of 284-296 mL, accounting for 59-61% of total MHV volume. There were no significant differences among the three types of liver partition evaluated. CONCLUSIONS: Our results demonstrate the reliability of virtual 3-dimensional reconstructions based on standard anatomical landmarks for both surgical planning and graft volume calculations. We believe our technique will help prevent small-for-size grafting and liver insufficiency resulting from inaccurate volumetric calculations. Our findings also support the observation that improved venous outflow might be achieved by including the MHV with the right liver graft without disadvantaging the left liver remnant. It is our hope that all these findings will translate into enhanced donor and recipient safety. PMID- 15816430 TI - A new "Perforator Grinder-Aspirator Apparatus (PGAA)" for the minimal access surgery of cystic liver hydatidosis. AB - Despite that minimal access surgery is a feasible, effective and viable option for radical surgical treatment of cystic hydatidosis, the laparoscopic approach is not widely accepted because of the risk of recurrence and the risk of severe complications, such as anaphylaxis and/or contamination of the peritoneal cavity related with the spillage. To overcome these difficulties, several techniques and devices are described in the literature. In our center (ISTEM), we have designed and developed a new generation "Perforator-Grinder-Aspirator Apparatus (PGAA: patent no. TR-2004/02916)" which evacuates cyst contents in a more secure, effective and easy way. By using PGAA, we are able to perform the laparoscopic approach while keeping the necessary pneumoperitoneum pressure to evacuate the cyst contents of liver hydatidosis securely and effectively, with wider indications even in difficult locations, after testing its safety and efficacy in some ex-vivo animal livers with hydatid cysts. PGAA is made up of four main units including puncture-irrigation needle, perforator-grinder pipe, aspiration cannula and, electric motor. One of the most important specialties of this new PGAA is having the automatically retractable rotary blade on the distal edge to grind and evacuate all the contents of the cavity including germinal layer and scolices. Having the retractable rotary blade allows the operator to avoid any intracystic parenchymal damage after perforating the outer layer. After taking out the perforator-grinder pipe, the aspiration cannula allows the laparoscopic visualization of the inner surface of the cystic cavity through its 10-mm internal diameter. In this article, the author describes the technical aspect and specifications of PGAA, and its applications in some cases with awkwardly located liver hydatid cysts, underlying its superiority over the classical aspirators and other devices. PMID- 15816431 TI - Successful preoperative treatment for hepatocellular carcinoma with tumor thrombus in the major portal branch by three-dimensional conformal radiation therapy--two case reports. AB - The prognosis of hepatocellular carcinoma (HCC) patients with tumor thrombus in the main portal branch (PVTT) is very poor and a standard treatment regimen for HCC with PVTT has not yet been established. Therefore, a new strategy is necessary to control the patients of HCC with PVTT. Conventional radiation therapy for HCC was not effective because of low liver tolerance. On the other hand, because three-dimensional conformal radiation therapy (3D-CRT) can focus high-dose irradiation on a small area, we tried it for patients with PVTT as the preoperative adjuvant treatment. 3D-CRT which targeted the PVTT was performed for two HCC patients with PVTT before radical hepatectomy. Treatment schedules of 3D CRT were as follows: in Case 1, the daily fraction size was 3 Gy, given five times per week, and the total dose was 39 Gy. In Case 2, the daily size was 6 Gy and the total dose was 60 Gy. Three or four weeks after 3D-CRT, the main tumor and PVTT were completely removed. On histological examination, the PVTT showed complete necrosis in Case 1, and 60% necrosis in Case 2. No serious complications occurred, and there have been no recurrences for more than one year after surgery in both cases. 3D-CRT targeting the PVTT may be promising as a preoperative adjuvant therapy for HCC with PVTT. PMID- 15816432 TI - Diagnosis and surgical treatment of primary hepatic cholangiocarcinoma. AB - BACKGROUND/AIMS: To summarize the experience in the diagnosis and surgical treatment of primary hepatic cholangiocarcinoma (PHCC). METHODOLOGY: Forty-seven PHCC patients, who had undergone surgical treatment in our hospital from 1970 to 1999, were retrospectively analyzed with their clinicopathological features, surgical treatments and prognoses. RESULTS: 1) PHCC patients lacked typical clinical manifestation, and some had an accompanying history of virus B hepatitis (36.2%, 17/47) and liver cirrhosis (46.8%, 23/47), and elevation of AFP (25.5%, 12/47). 2) The detection rates of BUS, CT, MRI and liver scintigraphy were 95.7% (45/47), 93.6% (44/47), 100% (23/23) and 100% (4/4), respectively. The lesions detected by those methods needed to differentiate with hepatocellular carcinoma. 3) The resection rate was 76.6% (36/47), the postoperative overall 1-, 3- and 5 year survival rates were 54.1% (24/47), 34.0% (16/47), and 29.8% (14/47) in all patients, respectively, including the 1-, 3- and 5-year survival rates were 83.3% (30/36), 50.0% (18/36), and 38.9% (14/36) in hepatectomy cases, respectively. 4) All of the four patients who survived more than 5 years, including small liver cancer (< or = 3 cm) three cases and encapsulated large nodular lesion one case, received curable resection and postoperative adjuvant therapies. CONCLUSIONS: PHCC patients can obtain a better result by early detection and curative resection with adjuvant therapies. PMID- 15816433 TI - Outcome and early complications of ERCP: a prospective single center study. AB - BACKGROUND/AIMS: ERCP has been used since 1968 both as a diagnostic and as a therapeutic procedure. In the last ten years several less invasive imaging methods have developed which allow visualization of bile ducts and pancreatic ducts; therefore ERCP has become mainly a therapeutic tool. The aim of the study was to prospectively evaluate the early outcome of ERCP when used essentially as a therapeutic tool. METHODOLOGY: All patients referred to our unit between April 1 1998 and March 31 2000 were included in the study. The indication for the procedure, number of cannulations, duration of procedure, performance of endoscopic sphinterotomy or pre-cut, insertion of stent and the final outcome were recorded. All patients were visited (in-patients) or contacted by phone (outpatients) 24 hours and one week after the procedure. If a complication occurred the patient was followed until it was resolved. RESULTS: 722 ERCP procedures were performed on 534 patients (59% females), mean age 63.4 years (range 3 to 98 years). The procedure was performed with a therapeutic intention in 95% of cases because of suspicion of: cholelithiasis (58%), tumor of pancreas/bile ducts (30%), post-cholecystectomy bile leak or bile duct stenosis (6%) or treatment of chronic pancreatitis (1%). The overall success rate of selective duct cannulation was 92%. Endoscopic sphincterotomy was performed in 375 procedures (52%), pre-cut in 152 (21%) and stent inserted in 180 (25%). Among the 701 procedures with complete information 76 complications occurred in 73 patients (10.8%): pancreatitis 4.3%, perforation 1.3%, sepsis 3.7%, bleeding 1.4%. Most of the complications (63/701, 9%) were mild to moderate. Procedure related mortality was 0.6% due to perforation (1 patient), and sepsis (3 patients). The main factors influencing the complication rate were the difficulty of the examination, performance of an invasive procedure or the operator. CONCLUSIONS: In the era of therapeutic ERCP, the risk of severe complication from the procedure is low--1.8% (including a mortality rate of 0.6%) when using the appropriate technique in experienced and skilled hands. PMID- 15816434 TI - Efficacy and safety of endoscopic retrograde cholangiopancreatography in patients with Billroth II anastomosis. AB - BACKGROUND/AIMS: Endoscopic retrograde cholangiopancreatography (ERCP) procedure is quite difficult to perform in patients with Billroth II anastomosis because of anatomical alterations. The aim of this study is to evaluate retrospectively the results of ERCP applications done in patients with Billroth II operation. METHODOLOGY: Out of the 1632 patients who underwent ERCP between 1992 and 2002, 27 (1.65%) had Billroth II operation. The records of these 27 patients were reviewed. Details noted included indications for ERCP, therapeutic interventions, causes of failure and complications. RESULTS: Out of the patients, 3 were female and 24 male (mean age 62+/-11). 26 patients had extrahepatic biliary obstruction. 1 patient had an external bile drain. The procedure was carried out 1-5 times (mean 1.5+/-1.1). Cannulation was achieved in 17 patients (62.96%). Out of the patients cannulated, 10 had choledocholithiasis, 4 malign choledochal stricture, 1 chronic pancreatitis, 1 bile leak and 1 periampullary tumor. Success rate of endoscopic treatment was 82.35% (14/17). Proximal migration of the stent and hemorrhage in gastric cardia were the complications observed in the distinct patients. CONCLUSIONS: ERCP procedure is quite an effective and safe method for diagnosis and treatment in patients with Billroth II anastomosis and extrahepatic cholestasis in spite of all difficulties. PMID- 15816435 TI - Mucosa-associated lymphoid tissue lymphoma of the extrahepatic bile duct. AB - Mucosa-associated lymphoid tissue lymphoma of the extrahepatic bile duct has not yet been reported. Much more common than this is secondary involvement of the extrahepatic bile duct in cases of disseminated lymphoma. A 59-year-old man manifesting jaundice was referred to our hospital. PTC revealed an extrahepatic bile duct stenosis from the hilum to the lower part of the choledochus. On the operative specimen, we examined L26/CD20, Bcl-2, UCHL-1/CD45RO, cyclin D1 and p53. Histologically, follicular colonization, centrocyte-like cells and lymphoepithelial lesion was observed. Tumor cells were positive for L26/CD20 and Bcl-2 and were negative for intracytoplasmic immunoglobulins, UCHL-1/CD45RO, cyclin D1 and p53. Pathological diagnosis was mucosa-associated lymphoid tissue lymphoma of the extrahepatic bile duct. The authors present herein the first case of mucosa-associated lymphoid tissue lymphoma of the extrahepatic bile duct. It was very difficult to distinguish from hilar cholangiocarcinoma clinically. Only incomplete stenosis of the bile duct and 18-F fluoro-2-deoxyglucose positron emission tomography (FDG-PET) could suggest this unusual clinical entity. PMID- 15816436 TI - A case of small cell carcinoma of the common bile duct. AB - Small cell carcinoma occasionally occurs in the gastrointestinal tract, but rarely in the biliary tract. We report a case of small cell carcinoma which occurred in the common bile duct. A 66-year-old female complained of epigastralgia and weight loss. Computed tomography and ultrasonography showed a mass near the pancreas head and dilatation of the intrahepatic bile ducts. Endoscopic nasobiliary drainage was undertaken, and it revealed obstruction of the common bile duct. The patient was diagnosed preoperatively as having extrahepatic bile duct cancer. Upon laparotomy, a tumor was found to be located in the middle common bile duct. Pylorus-preserving pancreaticoduodenectomy was performed. The main trunk of the portal vein and the right hepatic artery were resected concomitantly because of tumor involvement. Postoperative pathological examination revealed well-differentiated papillary adenocarcinoma on the surface of the bile duct lumen, but a large part of the extraductal component was small cell carcinoma. Upon immunohistochemical examination, synaptophysin and chromogranin A were found to be focally positive in small cell carcinoma, but negative for L-26 and CEA. The patient then underwent two postoperative courses of systemic chemotherapy. Nevertheless, she died of cancer recurrence eight months after the operation, which showed that the tumor had a highly lethal nature, with rapid and widespread dissemination. Further therapeutic trials are needed to improve survival in such cases. PMID- 15816437 TI - Intraoperative radiofrequency ablation: intraductal cooling of the main bile ducts for the prevention of heat damage. A case report. AB - Intraoperative radiofrequency (RF) ablation has recently become a valid procedure in the treatment of liver primary or secondary malignancies in selected cases. This procedure can be used alone or in association with surgery in treating lesions not considered for curative surgery. Heat damage of the main bile ducts represents nowadays a limitation in the use of radiofrequency ablation of hepatic lesions. Up to now, in tumors proximal to a main bile duct or to the biliary bifurcation this procedure has been contraindicated. In order to preserve these structures from heat damage, a new technique consisting of intraductal cooling of the bile ducts has recently been introduced. We report a case of successful experience with this technique. PMID- 15816438 TI - Anicteric early bile duct carcinoma detection with magnetic resonance cholangiopancreatography. AB - The poor prognosis of extrahepatic bile duct carcinoma makes early detection and diagnosis essential for positive patient outcomes. We describe 2 cases of jaundice-free early extrahepatic bile duct carcinoma detected by magnetic resonance cholangiopancreatography. Extrahepatic bile duct carcinoma was discovered incidentally in patient 1 by magnetic resonance cholangiopancreatography during evaluation of a gallbladder stone. In patient 2, extrahepatic bile duct carcinoma was found during a routine health maintenance exam. Both patients underwent radical surgical intervention. Both patient 1 and 2 have remained in good health for over one year, 3.5 and one year, respectively, and have not exhibited any signs or symptoms of relapse or cancer recurrence. Based on these cases, it appears that magnetic resonance cholangiopancreatography can play a significant role in the early detection of extrahepatic bile duct carcinoma and improve disease prognosis. PMID- 15816439 TI - Postoperative survival in intrahepatic cholangiocarcinoma showing intraductal growth. AB - Clinicopathologic variables favoring recurrence after hepatic resection for intrahepatic cholangiocarcinoma showing intraductal growth remain unclear. We investigated various clinicopathologic features in three patients who underwent resection for this type of intrahepatic cholangiocarcinoma. All underwent extended left hepatectomy plus resection of the caudate lobe and lymph node dissection. Lymph nodes showed no pathologic involvement. Although no cancer cells were seen in the mucosal layer by intraoperative pathologic examination at the bile duct stump in any patient, pathologic examination of resected specimens showed cancer cells invading beyond the mucosal layer in connective tissues surrounding the bile duct stump (interstitial invasion) of the Glisson's sheath in 2 patients. One of them died of cancer recurrence near the bile duct stump, while the third patient, without interstitial invasion, has survived for 10.6 years. In the intraductal growth type of intrahepatic cholangiocarcinoma, absence of cancer cells should be confirmed by intraoperative pathologic examination of not only the mucosal layer of the bile duct but also the connective tissue surrounding the bile duct, since interstitial invasion may be a risk factor for cancer recurrence. PMID- 15816440 TI - A new technique for the palliative treatment of high bile duct cancer. AB - In case of advanced unresectable hilar cholangiocarcinoma palliative procedures can offer a good quality of life. Palliative techniques include surgical intubation with a U tube or transhepatic ring catheters, nonoperative intubation with an endoprosthesis and biliary-enteric bypass techniques. We present a new surgical procedure for the palliative treatment of high bile duct cancer, in which the gallbladder is used as a conduit for a biliary-enteric bypass. Hepatico cholecysto-jejunostomy is a safe, rapid and easy method that could be alternatively and successfully used for surgical palliation of unresectable hilar cholangiocarcinoma. PMID- 15816441 TI - Curative reoperation for recurrent cancer of the extrahepatic bile duct: report of two cases. AB - Local recurrence, following a resection for cancer of the extrahepatic bile duct, is usually incurable with second curative surgery being almost impossible. To determine the feasibility and significance of second curative surgery, our experiences are presented in this study. The medical records and clinical outcomes of two patients that underwent a re-resection for recurrent cancer of the extrahepatic bile duct were retrospectively reviewed. A 50-year-old female patient that had a recurrent disease at the intrahepatic and intrapancreatic bile duct, 66 months after a segmental resection of the bile duct for common bile duct (CBD) cancer, underwent a hepatopancreatoduodenectomy. A 29-year-old female patient had a recurrent tumor mass in the distal CBD, 28 months after a right hemihepatectomy and Roux-en-Y hepaticojejenostomy for a type IIIa Klatskin tumor, and underwent a segmental resection of the bile duct. The gross type of the above two cases was a papillary tumor. There was no operative mortality or morbidity. All patients are still alive after 46 and 9 months, respectively, without recurrence after the reoperation. It is concluded that a surgical re-resection is possible in selected patients with recurrent bile duct cancer, mostly of the papillary type. A primary operation for bile duct cancer should be performed with a wide surgical margin, and secondary curative surgery should be considered whenever possible in cases of recurrence. PMID- 15816442 TI - Multimodality approach towards advanced rectal carcinoma. "Its efficacy and safety". AB - BACKGROUND/AIMS: To evaluate the efficacy and safety of multimodality approach towards advanced rectal cancer. METHODOLOGY: In a prospective, randomized trial, two hundred and ten patients with advanced carcinoma of the rectum were studied. Patients were randomly divided in two groups A and B. Patients in group A underwent total mesorectal excision (TME) with adjuvant systemic chemotherapy and radiotherapy. Group B patients underwent TME with adjuvant stop-flow perfusion along with systemic chemotherapy RESULTS: The five-year survival for Group A patients was 72% and for Group B patients the overall five-year survival was 92% and the disease-free survival was 87%. CONCLUSIONS: Multimodality approach to advanced rectal cancer leads to substantial prolongation of survival and optimization of quality of life. PMID- 15816443 TI - Factors predisposing to poorly differentiated hepatocellular carcinoma and its recurrence. AB - BACKGROUND/AIMS: We investigated the clinical factors predisposing moderately or poorly differentiated hepatocellular carcinoma and analyzed which clinical and histological factors are associated with poorly differentiated hepatocellular carcinoma (HCC) recurrence. METHODOLOGY: Percutaneous fine-needle biopsy was taken from the liver tumor of 191 consecutive patients between January 1994 and September 1996. The histological degree of differentiation of hepatocellular carcinoma at the first time of initial treatment and at the time of second recurrence was classified according to the criteria of Edmondson and Steiner. RESULTS: At the time of the first therapy, 86 patients, 81, 24, and 0 patients had liver tumors classified as Edmondson (Ed), 1, 2, 3, and 4, respectively. The prognosis of patients with Ed-3/4 HCC was worse than and the tumor sizes were larger than that of Ed-1/2 HCC patients. Of the 167 patients classified as Ed-1/2 at the time of first therapy, HCC recurred in 95 of the patients during the mean follow-up period of 3.4 years. Multivariate analysis revealed that only tumor size (P=0.035) and TACE therapy (P=0.0009) were independently significant factors in predicting future Ed-3/4 or multiple HCC recurrence. CONCLUSIONS: Tumor size and TACE therapy were clinical predisposing factors for Ed-3/4 or multiple HCC recurrences. PMID- 15816445 TI - Endoscopic findings potentially predictive of gastric cancer in borderline lesions diagnosed by forceps biopsy. AB - BACKGROUND/AIMS: When a benign-malignant borderline lesion is diagnosed by the usual small gastric biopsy, there is sometimes difficulty in making a clinical decision. To clarify potentially useful findings to predict the existence of gastric cancer in borderline lesions diagnosed by forceps biopsy, we retrospectively analyzed endoscopic features. METHODOLOGY: We diagnosed 68 consecutive gastric benign-malignant borderline lesions (57 cases) by forceps biopsy and endoscopically resected them. The final diagnosis for 24 lesions (35.3%) was adenocarcinoma (adenocarcinoma group), and for 40 lesions (58.8%) was adenoma (adenoma group). Comparison with endoscopic findings for the groups was carried out using digitally filed endoscopic photos. RESULTS: We found six endoscopic findings (distal location, reddish surface color, lack of smoothness, lack of glossiness, focal roughness, and focal redness) having statistically significant relationships with adenocarcinoma at the final pathological diagnosis. In multivariate analysis, focal redness (p<0.01) and lack of glossiness (p<0.05) were found to have a significant relationship to gastric cancer. CONCLUSIONS: Endoscopic findings such as focal redness and lack of glossiness were potentially predictive of gastric cancer in borderline lesions diagnosed by forceps biopsy. PMID- 15816444 TI - MUC1 overexpression is the most reliable marker of invasive carcinoma in intraductal papillary-mucinous tumor (IPMT). AB - BACKGROUND/AIMS: To clarify the development of pancreatic cancer we performed immunohistochemical analysis of the presence of the major apomucin and cell-cycle regulatory proteins using the tissues of IPMT and ductal adenocarcinoma (DC) of the pancreas. METHODOLOGY: Formalin-fixed and paraffin-embedded tissues of 24 IPMT and 21 DC cases were subjected to immunohistochemical staining for MUC1, MUC2, p16, p53 and DPC4. According to the WHO classification, there were 10 intraductal papillary-mucinous adenomas (IPMA); 3 borderline intraductal papillary-mucinous neoplasms (IPMB); 4 intraductal papillary-mucinous carcinomas (IPMC), non-invasive type (nIPMC); 4 IPMCs with invasive muci nous carcinoma (IPMC/muc); and 3 IPMCs with invasive tubular adenocarcinoma (IPMC/tub). RESULTS: MUC1 expression was seen in 6 of 7 invasive IPMCs (86%) and in all DCs (100%). MUC2 was only seen in non-invasive IPMT and in a part of IPMC/muc. p53 nuclear staining was positive only in 3 of 7 invasive IPMCs (43%) and 9 of 21 DCs (43%). DPC4 nuclear expression was positive in almost all cases of non-invasive IPMT, but negative or reduced in 4 of 7 invasive IPMCs (57%), and 14 of 21 DCs (67%). CONCLUSIONS: MUC1 overexpression is considered to be the most sensitive and specific marker of invasive carcinoma, followed by DPC4 and p53 with less sensitivity. PMID- 15816446 TI - Gastrointestinal stromal tumors (GIST). AB - BACKGROUND/AIMS: Gastrointestinal stromal tumors (GIST) are the most frequent non epithelial tumors of the alimentary tract. The interstitial cells of Cajal or more primitive progenitor mesenchymal cells are suggested as their cells of origin. GIST's occur throughout the gastrointestinal tract but are generally located in the stomach and the intestine. About 70% of GIST's are immunohistochemically positive for CD34 and more than 90% for c-kit protein (CD117). About two thirds of GIST's are malignant. The tumor size, mitotic rate, cellularity and nuclear pleomorphism are the most important parameters characterizing the biological behavior of tumors. The diagnostic procedures are similar to those of other gastrointestinal neoplasms but only a half of the patients will have correct preoperative histological diagnosis. GIST's can be cured only by surgery. The procedure of choice, if possible, is resection without extended lymphadenectomy. Radiation and chemotherapy are generally ineffective. METHODOLOGY: 22 patients were operated on for GISTs in our department between 1996 and 2003. RESULTS: All but one proved to be benign. The patients were all asymptomatic after the operation except the malignant case. In his case an irresectable local recurrence developed later. CONCLUSIONS: GIST is a rare neoplasm of the GI tract. The only possibility for treatment of GISTs is surgical removal. PMID- 15816447 TI - Capsule endoscopy; the cumulative experience from its use in 193 patients with suspected small bowel disease. AB - BACKGROUND/AIMS: This study presents our cumulative clinical experience from the use of wireless capsule endoscopy (WCE) in a large series of Greek patients with suspected small bowel (SB) diseases. METHODOLOGY: Over an 18-month period, 193 patients were evaluated with WCE for suspected SB diseases [108 obscure gastrointestinal (GI) bleeding, 32 chronic diarrhea, 22 suspected and 6 diagnosed Crohn's disease, 16 abdominal pain, 4 refractory celiac disease, 3 polyposis syndromes, 1 Rendu-Osler-Weber disease and 1 Behcet disease]. All patients had undergone upper GI endoscopy and total colonoscopy. WCE findings were characterized as specific or non-specific, depending on whether the patient's signs and symptoms could be sufficiently attributed to them or not. RESULTS: One or more abnormal findings were detected in 161/193 (83%) patients; these were classified as specific in 91/193 (47%). The diagnostic yield of WCE (in terms of specific findings) was significantly higher when evaluating patients with obscure GI bleeding, compared to chronic diarrhea (52% vs. 25%, respectively, p=0.013, 95% CI: 1.33-7.83). Among obscure GI bleeders, specific findings were significantly more in the group of overt rather than occult bleeders (66% vs. 42% respectively, p=0.026, 95% CI: 1.19-5.88). In patients referred for diagnosed or suspected Crohn's disease, WCE findings were compatible with the diagnosis in 5/6 cases (83%) and in 8/22 cases (36%), respectively. In 9 more patients from other groups with a negative previous diagnostic work-up, WCE helped in diagnosing Crohn's disease. Among 16 patients evaluated for abdominal pain, specific findings were identified only in 1 (6%). CONCLUSIONS: WCE seems to be a very useful diagnostic tool in the evaluation of GI bleeding of obscure origin (in both overt and occult bleeders), as well as suspected Crohn's disease. In other indications, such as unexplained chronic diarrhea and abdominal pain, it completes the traditional work-up, but abnormal findings need to be better delineated before WCE can be widely recommended in these cases. PMID- 15816448 TI - Brain MRI findings in cirrhotic patients: correlation with Doppler US hepatic flow parameters. AB - BACKGROUND/AIMS: To determine the relationship of basal ganglia alterations demonstrated on brain MRI and clinical presentation, evaluated with Child-Pugh score, using Color Doppler Ultrasonography parameters of portal hemodynamics in cirrhotic patients with signs of portal hypertension. METHODOLOGY: Twenty randomly selected cirrhotic patients (16 males and 4 females with a mean age of 63.6+/-8.9 years) classified according to Child-Pugh score (grade A=16 patients, grade B=4), were submitted to Color Doppler US evaluation of hepatic flow and brain MRI. The obtained flow parameters were: PVD, PVCSA, MPVF, MPVFV, HAD, MHAF, hepatic artery RI, CI and MHI. The basal ganglia signal intensity, on T1-weighted images on brain MRI was evaluated both qualitatively (normal, mild, moderate and severe) and quantitatively with the ROI method. RESULTS: Among Color Doppler parameters, only the CI exhibited a modest correlation with the Child's clinical score (p for linear trend by ANOVA <0.05). Subjective MRI grading demonstrated an excellent correlation with the qualitative assessment of signal density (Spearman p=0.95, p<0.01). When signal intensity on T1-weighted images was analyzed as a continuous variable with normal distribution and ultrasonographic parameters as possible determinants, the portal vein diameter (PVD) and consequently the cross sectional area (PVCSA) emerged as the sole predictor of MR signal intensity (Pearson's r=0.58, p=0.01). CONCLUSIONS: Although this is a preliminary study and further research should be performed including a larger number of patients, it suggests that Color Doppler US might play a prognostic role in predicting hepatocellular dysfunction and hepatic encephalopathy. PMID- 15816449 TI - Huge ovarian metastasis from jejunal cancer occurring immediately after initial operation. AB - Correct preoperative diagnosis of jejunal cancer is difficult, and there are no clear guidelines for the postoperative follow-up to monitor the recurrence of jejunal cancer. This report describes asynchronous ovarian metastasis from jejunal cancer occurring after initial operation. The patient was a 68-year-old postmenopausal woman with a preoperatively correctly diagnosed jejunal cancer. Partial resection of the jejunum with regional lymph node dissection was performed, with both ovaries intact at the first operation. Three months later, vaginal bleeding and a large metastasis in the left ovary were detected, for which a left oophorectomy was performed. Peritoneal dissemination was also detected perioperatively, so that chemotherapy with 5-FU/leukovoline was started. The patient is still alive more than 12 months after the most recent operation. Our case demonstrates that it is necessary to pay careful attention to synchronous or non-synchronous metastases to the ovaries after operation for jejunal cancer in females. Consideration should also be given to the efficacy of oophorectomy for ovarian metastases and of intensive chemotherapy combined with jejunal resection for carcinoma because of the poor prognosis of these treatment modalities. PMID- 15816450 TI - Endoscopic piecemeal mucosal resection of large colorectal tumors. AB - BACKGROUND/AIMS: Since endoscopic en bloc resection of large and sessile tumors is technically difficult, endoscopic en bloc piecemeal mucosal resection (EPMR) is usually chosen for resection of such tumors. Tumors resected by EPMR are, however, difficult to evaluate histologically. The aim of this study was to evaluate the safety and effectiveness of EPMR. METHODOLOGY: We removed 30 large colorectal tumors in 30 patients by EPMR between 1992-2000. Endoscopic examination was repeated at 3, 6 and 12 months and later on after initial endoscopic resection. Patients in whom no residual tumor was found by both endoscopic and histologic examination were considered to be "cured". RESULTS: Histological examination of the resected tumor tissues revealed malignancy in 43.3% (13/30). Three patients had invasive malignant tumors and underwent surgery. Following complete endoscopic resection, recurrences were observed in 2 patients with benign tumors, which were resected by additional endoscopic resection. All patients including the two with non-invasive malignant tumors remain free from recurrence during a mean follow-up period of 45.2 months (range, 3-104 months). Bleeding was the only complication and was seen in one patient (3.3%; 1/30), which was treated by endoscopic clipping. CONCLUSIONS: EPMR of benign or non-invasive large malignant tumors is a safe and effective procedure. Complete excision of large, sessile and non-invasive tumors is possible, although complete removal by EPMR cannot be verified histologically. PMID- 15816451 TI - Polyamines levels in colorectal cancer: new markers? AB - BACKGROUND/AIMS: The aim of this study is to evaluate free polyamine and acetyl derivate levels in colorectal cancer, precancerous lesions and a control group in order to investigate if there are different concentrations between neoplastic and non-neoplastic lesions. METHODOLOGY: For this study, in the Department of Surgical Sciences of Catania University we observed 62 patients affected by colorectal cancer, 32 affected by precancerous lesions and 20 used as the control group. In each patient we performed an endoscopic biopsy on the lesion, one 5 cm to the lesion and one 10 cm to the lesion. In each biopsy were detected polyamines concentrations. RESULTS: We found a significant increment of free and acetylated polyamines levels in neoplastic patients in comparison with non neoplastic. Polyamines levels are increased at the site of cancer in comparison of adjacent colonic mucosa. In addition polyamines levels are increased in normal colon mucosa from cancer patients compared with the mucosa of control patients. CONCLUSIONS: Significant increment of polyamines levels at the site of cancer in comparison of non-neoplastic mucosa and adjacent colonic mucosa give them the possibility to occupy an important role in early detection of colon cancer by acting like oncological markers. PMID- 15816452 TI - Clinicopathologic differences among subtypes of serrated adenomas of the colorectum. AB - BACKGROUND/AIMS: Serrated adenomas (SAs) of the colorectum can be broadly divided into two subtypes: type I more closely mimicking hyperplastic polyps, and type II unequivocal traditional adenomas. The aim of this study was to clarify their differential clinicopathologic and colonoscopic features. METHODOLOGY: A total of 127 SAs (53 type I, 52 type II and 22 admixed type I+II) were investigated and colonoscopic surface patterns were divided into three categories: speckled, granular and cerebriform. RESULTS: The cerebriform pattern was most frequently observed in all SA types. Types I+II (median size, 7.5 mm) or type II SAs (median size, 10 mm) were generally sessile or pedunculated polyps in the rectosigmoid colon whereas some type I lesions (median size, 5 mm) demonstrated a flat elevated morphology and were found in the ascending colon and cecum. Co-existing (2/127: 1.6%) invasive carcinomas were only detected with type II SAs. In contrast, synchronous invasive carcinomas distant from SAs were more frequently observed with type I (31%) than types I+II (5%) or II (12%). CONCLUSIONS: Clinicopathologic differences are apparent among the types of SAs. A type II SA invasive carcinoma sequence might exist. We stress recognition of type I SA as a neoplastic, rather than a hyperplastic lesion, often accompanying invasive carcinomas at a distance from the SA. PMID- 15816453 TI - The administration of cisapride as an adjuvant to PEG-electrolyte solution for colonic cleansing: a double-blind randomized study. AB - BACKGROUND/AIMS: Traditional bowel preparation before colonoscopy involves lavage with approximately 4L of polyethylene glycol (PEG)-electrolyte solution. Only a few studies have been published evaluating the use of cisapride in routine bowel preparation. METHODOLOGY: We conducted a blinded, placebo-controlled trial with the prokinetic agent, cisapride, in addition to standard PEG-electrolyte lavage. Of 115 patients undergoing colonoscopy, 58 were randomized (double-blind) to PEG plus cisapride (10 mg per os thrice per day three days before the procedure and one 10-mg dose on the morning of the procedure) and 57 to PEG plus a placebo of identical appearance. The adequacy of the preparation was scored on a four-point grading scale for each anatomic-segment and for the overall impression. A questionnaire was also used to assess each patient's symptoms during lavage. RESULTS: The difference in the overall score between the two groups was not significant (p=0.21). The quality of bowel preparation was significantly better in transverse (p=0.001), ascending (p=0.0053), and cecum (p=0.0001) in the cisapride group than in the placebo group. The differences in symptoms scores between the two groups were not significant in nausea, abdominal cramps and bloating but there was improvement in symptom score of vomiting in cisapride group (p=0.0422). CONCLUSIONS: The administration of cisapride to patients undergoing colonic lavage may be an effective adjuvant to PEG-electrolyte solution particularly with respect to increase patient acceptability. PMID- 15816454 TI - Postsurgical surveillance for recurrence of UICC stage I colorectal carcinoma: is follow-up by CEA justified? AB - BACKGROUND/AIMS: This study was undertaken to investigate whether it will be possible to reduce the times and types of postoperative examinations for surveillance in patients with UICC stage I colorectal carcinoma. In addition, the value of CEA in postoperative surveillance is discussed. METHODOLOGY: A review was performed of 541 patients who underwent curative resection for UICC stage I colorectal carcinoma between January, 1985 and December, 1998. Periodic check-up was routinely conducted to identify recurrence. RESULTS: The median follow-up was 82 months. The recurrence rate was 2.9% in the UICC stage Ia (pT1N0M0) group, and 5.6% in the Ib (pT2N0M0) group. Cancer-specific survival rates at 5 years were 99.3% and 97.6%, respectively (p=0.0354). Recurrences occurred more frequently in patients with lower rectal carcinoma (p=0.0415). Curative-intent salvage surgery was performed in 61.9% (13/21) for recurrent lesions. Between the patients who were CEA positive (13/21; 61.9%) and those who were CEA negative at the time of recurrence, there was no significant difference in the prognosis. CONCLUSIONS: The incidence of recurrence was low after curative surgery in patients with UICC stage I colorectal carcinoma, and it is therefore possible to reduce times and types of postoperative examinations. CEA measurement alone appears to be sufficient. PMID- 15816456 TI - Local recurrence and survival rates after extended systematic lymph-node dissection for surgical treatment of rectal cancer. AB - BACKGROUND/AIMS: Extended systematic lymph-node dissection (ESLND) is a surgical procedure aimed at decreasing the local recurrence rate of rectal cancer and increasing the survival rate. However, it is criticized because it has not shown the expected effects on survival, and it has been shown to increase the proportion of complications in rectal cancer surgery. This study was designed to determine incidence and patterns of recurrence after curative resection with or without ESLND for rectal cancer. METHODOLOGY: A total of 184 patients with rectal cancer were reviewed with respect to surgical procedures, local recurrence and survival rates. RESULTS: 170 of 184 patients with rectal cancer were administered curative surgical resection. ESLND was added to the surgical procedure of 24 of these 170 patients. The local recurrence rate of the patients who did not receive lymph-node dissection was 15%, and the survival rate over 5 years was 58.9%. The local recurrence rate of the patients receiving ESLND was 12.5%, and the survival rate over 5 years was found to be 55.7% (p>0.05). CONCLUSIONS: Because ESLND is a procedure added on to curative resection in the surgical treatment of rectal cancer, it increases the general anesthesia and length of surgery, and it is possible that some complications due to the operation itself may arise. In the current study and in a large amount of research in the literature, a statistically significant effect on the survival rate has not been found. In conclusion, the opinion has been reached that ESLND does not have an important benefit in the curative surgical treatment of rectal cancer. PMID- 15816455 TI - Correlation between hMLH1/hMSH2 and p53 protein expression in sporadic colorectal cancer. AB - BACKGROUND/AIMS: The aim of this study was to analyze expression of hMLH1 and hMSH2 mismatch repair proteins in terms of p53 protein expression and clinicopathological parameters in sporadic colorectal cancer. METHODOLOGY: Four hundred and two cases of curative colorectal surgery for primary colorectal cancer were included in this study (patients with a familial history of colorectal cancer and familial adenomatous polyposis were not included). Clinicopathological parameters were reviewed retrospectively. HMLH1, hMSH2 and p53 protein expression in tumor tissue sections was determined using immunohistochemical staining with specific monoclonal antibodies. RESULTS: Of the 402 cases, immunohistochemical analysis showed 35 (8.7%) had loss of expression of hMLH1, 19 (4.7%) had loss of expression of hMSH2, and three cases (0.7%) had loss of expression of both proteins. Multivariate analysis showed that early age of onset (p=0.023), right side dominance (p<0.001) and poorly differentiated or mucinous cell type (p<0.001) were associated with loss of expression of hMLH1 or hMSH2. Loss of expression of hMLH1 or hMSH2 correlated with low p53 expression (p<0.001). In terms of clinicopathological parameters, p53 expression was associated only with hMLH1 or hMSH2 expression. CONCLUSIONS: Colorectal cancers not expressing hMLH1 or hMSH2 may have distinct features from those expressing these mismatch repair proteins. p53 expression appears to be implicated in a compensatory pathway with mismatch repair proteins. PMID- 15816457 TI - Transanal local wide excision for rectal adenocarcinoma. AB - BACKGROUND/AIMS: Abdominoperineal resection is associated with high morbidity and mortality, and sphincter preservation is the aim for the patient. Transanal local wide excision of highly selected rectal cancers is an acceptable alternative to radical surgery. METHODOLOGY: This retrospective study of 18 patients with rectal cancer treated with transanal local wide excision at our hospital during a 6-year period (from 1995 to 2001) is discussed. RESULTS: Tumor size ranged from 1 to 6 cm (mean, 2.23 cm). All resection margins were free of tumors. There was no surgical mortality or morbidity. Median follow-up period was 17.9 months. Among 18 patients, 12 patients received radiotherapy and chemoradiotherapy as a postoperative adjuvant treatment. There was one local recurrence with liver metastasis noted within one year after the operation. The 1-yr, 2-yr and 5-yr disease-free survival rate was 92%. CONCLUSIONS: Transanal local wide excision for rectal cancer, when combined with selected chemotherapy or radiotherapy, results in good local-regional control in our series. A Good long-term survival rate was also proven by presented articles. This approach can be safely applied to more advanced tumors (T3 lesion) under accurate pre-op staging, aggressive postoperative adjuvant therapy and careful regular follow-up. PMID- 15816459 TI - The superior mesenteric artery syndrome; an unusual cause of vomiting. AB - A 29-year-old female presented with anorexia, vomiting and weight loss. A diagnosis of superior mesenteric artery syndrome was made based upon contrast duodenography and then at laparotomy when the patient was successfully treated with a duodenojejunostomy. Superior mesenteric artery syndrome is a rare cause of vomiting not detectable by endoscopy, but is eminently amenable to treatment. PMID- 15816458 TI - The role of interleukin-1 beta (IL-1beta) in the normal human colon in vitro. AB - BACKGROUND/AIMS: The possibility that interleukin-1 beta (IL-1beta) is a neuromodulator of the non-adrenergic non-cholinergic (NANC) inhibitory nerves which may be mediated by nitric oxide (NO) was recently reported from animal experiments. To clarify the physiological significance of the relationship between IL-1beta and NO in the normal human colon, enteric nervous responses to IL-1beta in the normal colon muscle strips were investigated. METHODOLOGY: Normal colon muscle strips derived from patients who underwent colon resection for left sided colon cancers (14 cases) were used. The subjects consisted of 8 men and 6 women, aged from 44 to 65 years with a mean age of 56.8 years. A mechanographic technique was used to evaluate in vitro colon muscle responses to IL-1beta of adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers and N(G)-nitro-L-arginine (L-NNA). RESULTS: IL-1beta concentration dependently caused a relaxation reaction before and after the blockade of the adrenergic and cholinergic nerves. The frequency of relaxation responses after blocking the adrenergic and cholinergic nerves was higher than that before blocking, but there was no significant difference between them. Both tetrodotoxin and L-NNA inhibited the relaxation reaction in response to IL-1beta in the human colon. CONCLUSIONS: These findings suggest that IL-1beta plays an important role in regulating relaxation of the normal human colon via nitregic nerves, and that NO plays a role as a neurotransmitter in the NANC inhibitory nerves. PMID- 15816460 TI - Effect of an aldose reductase inhibitor on esophageal dysfunction in diabetic patients. AB - BACKGROUND/AIMS: Disorders of the digestive tract in diabetic patients are mainly ascribed to disorders of the vagus nerve. Although aldose reductase inhibitors (ARIs) have been shown to be effective against diabetic peripheral neuropathy, their effectiveness on diabetic digestive neuropathy remains to be evaluated. The aim of the present study is to examine the effect of an ARI on the esophageal dysfunction in diabetic patients by monitoring pH and motility of the esophagus. METHODOLOGY: Eight type 2 diabetic patients with peripheral neuropathy were administered with the ARI epalrestat (150 mg/day) for 90 days, and esophageal pH and motility were monitored before and after the ARI treatment. RESULTS: Parameters related to the gastroesophageal acid reflux and the esophageal motility, such as % time of pH<4, DeMeester score, duration of the longest reflux episode, reflux episodes longer than 5 min, ratios of peristaltic waves with the amplitude greater than 25 mmHg and ratios of effective peristalsis were remarkably improved by the ARI treatment. CONCLUSIONS: Because the present study clearly demonstrated the effectiveness of an ARI on the esophageal dysfunction in diabetic patients, ARI may be useful for the treatment of diabetic digestive disorders. PMID- 15816461 TI - Pneumatic balloon dilatation in primary achalasia: the long-term follow-up results. AB - BACKGROUND/AIMS: Pneumatic dilatation is a safe and most effective treatment for achalasia. We analyzed the long-term results of pneumatic dilatation in primary achalasia by objective and subjective findings. METHODOLOGY: Pneumatic dilatation was performed in patients that were diagnosed with primary achalasia in our manometry laboratory between 1993-1999 years. We evaluated patients with clinical, radiologic, endoscopic and manometric results before treatment. Mean esophageal diameters on the level of the lower esophageal sphincter and middle esophagus were measured by barium esophagograms. The patients were clinically reevaluated after one week and barium esophagograms were repeated one month later after dilatation. Clinical examination, endoscopy and manometry were done at 1, 3, 6 and 12 months and repeated yearly for follow-up period. A statistical comparison of pre- and posttreatment on the frequency of dysphagia, radiological diameter of the esophagus and manometric data was performed using unpaired t tests and chi2 tests. RESULTS: Pneumatic dilatation was performed on 50 adult patients with a mean age 41.42+/-18.07 years. A single dilatation was successful in forty patients (80%) and two to three dilatations were performed in ten (20%) patients. The median number of dilatations was 1.26. In the postdilatation period, mean short-term (< 1 year) and long-term (2-7 years) clinical improvement was 82.8% and 66.85% respectively. The mean diameter of the esophagus was regressed to 26.51+/-7.69 mm from 36.66+/-11.23 mm (p<0.001) and the mean diameter of the lower esophageal sphincter was increased to 8.38+/-3.12 mm from 2.58+/-1.13 mm (p<0.001) with pneumatic dilatation. The mean pretreatment pressure of lower esophageal sphincter was 41.14+/-11.34 mmHg and these values were 18.79+/-7.85 mmHg (p<0.001), 13.18+/-9.53 mmHg (p<0.001) in the 1st, and 5th years of the posttreatment period, respectively. The mean pressure of the lower esophageal sphincter was 31.78+/-8.91 mmHg in nonresponder patients during the posttreatment period; there was no significant difference prior to pneumatic dilatation (p>0.1). Surgical operation was performed on 5 patients (10%), who had no benefit from pneumatic dilatation. CONCLUSIONS: Pneumatic dilatation is an effective procedure in the treatment of primary achalasia during the short- and long-term period. Treatment evaluation can possibly be made objectively with radiographic and manometric alterations of esophagus that occurred after pneumatic dilatation. PMID- 15816462 TI - Is preoperative methylprednisolone beneficial for patients undergoing esophagectomy? AB - BACKGROUND/AIMS: This study examined whether or not preoperative administration of methylprednisolone reduces perioperative systemic inflammatory response and thus can offer clinical benefit to patients. METHODOLOGY: Forty patients with thoracic esophageal cancer who were scheduled for esophagectomy with lymphadenectomy were randomly allocated to either of the following two groups: 20 patients received methylprednisolone (500 mg/body) in saline 2 hours before surgery and another 20 patients received a placebo (saline alone). The effects on the early clinical course were assessed by rectal intramucosal pH (pHi) as a marker of systemic oxygenation, water balance, serum cytokine levels and the incidence of postoperative complications. The effects on the late clinical course were examined by comparing patient survival time. RESULTS: Rectal pHi value, water balance, blood balance and body weight change did not significantly differ between the two groups. Serum proinflammatory cytokine (interleukin-6 and interleukin-8) levels were significantly higher in the control group than the methylprednisolone group 2 hours after surgery. Serum C-reactive protein was also significantly higher in the control group on postoperative day 2 and 3. No significant differences were seen in the incidence of postoperative complications. Survival curves for the two groups did not statistically differ. CONCLUSIONS: Preoperative methylprednisolone significantly reduces inflammatory cytokine response immediately after surgery for esophageal cancer, but its clinical benefit remains unclear. PMID- 15816463 TI - Relationship between glucose transporter, hexokinase and FDG-PET in esophageal cancer. AB - BACKGROUND/AIMS: 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) has been established as a powerful diagnosing modality in clinical oncology. FDG accumulation has been demonstrated to correlate with hexokinase activity. However, recent reports suggest that glucose transporters participate in FDG accumulation. The aim of this study is to evaluate glucose transporter and hexokinase expression and clarify the relationship between them and FDG accumulation. METHODOLOGY: FDG-PET was performed in 72 preoperative patients with esophageal cancer. The ratios of tumor radioactivity to plasma radioactivity (Ci/Cp values) were obtained 60 minutes after administration. We studied the expressions of glucose transporter 1 (Glut1) and type-II hexokinase (HK-II) by immunohistochemical analysis of the resected specimen. The percentages of cells expressing Glut1 and HK-II were scored on a 5-point scale (1=0-20%, 2=20-40%, 3=40-60%, 4=60-80%, 5=80-100%). Then the 3 scores obtained from 3 counting trials were averaged to give the Glut-index and HK-index. RESULTS: All esophageal cancers showed marked FDG accumulation. All 72 cancers expressed Glut1 and 71 of 72 cancers expressed HK-II. The Glut-index had a weak correlation with the Ci/Cp value (not significant). The HK-index had a close positive correlation with the Ci/Cp value (p<0.005). CONCLUSIONS: FDG accumulation correlates more with type-II hexokinase expression than with glucose transporter 1 expression. PMID- 15816464 TI - Detection of gastrointestinal and abdominal infections by 99mTc-ciprofloxacin. AB - BACKGROUND/AIMS: The aim of the study is detection and evaluation of the abdominal and gastrointestinal infective foci using 99mTc-ciprofloxacin (Laboratory for radioactive isotopes, Vinca). METHODOLOGY: In total 21 patients with clinical suspicion on abdominal or gastrointestinal infection were investigated. In all the patients, planar liver/spleen scintigraphy was performed. Ciprofloxacin chloride (3.5 mg) was mixed with 555 MBq of 99mTc in 3 mL of physiological solution and incubated for 20 min. After slow i.v. injection in a cubital vein, dynamic acquisition (1 f/min) was performed during the first 60 min in the position of interest, followed by static acquisition (500,000 imp) anterior and posterior view, abdomen and pelvis after 1 h and 4 h in all patients. When necessary, additional scintigrams were acquired after 24 h. In all the patients with negative or equivocal findings of planar scintigraphy, emission computerized tomography (SPECT) was performed (60 positions, 6 degrees). Interpretation was made by three independent observers. Additional data were provided using clinical findings, ultrasonography, computed tomography and magnetic resonance imaging, laboratory analyses, and surgical or microbiological confirmation of infection. RESULTS: There were eleven true-positive findings, seven true negative, two were false negative while one was false positive due to intestinal obstruction. Sensitivity was 79%, specificity 91%, positive predictive value 92%, negative predictive value 77%, accuracy 84%. CONCLUSIONS: According to our results, scintigraphy with radiolabeled ciprofloxacin is a useful method for detection and assessment of exact localization of abdominal and gastrointestinal infections. PMID- 15816465 TI - Epidemiological aspects of endemic upper gastrointestinal cancers in eastern Turkey. AB - BACKGROUND/AIMS: The aim of our study was to present the epidemiological aspects of endemic upper gastrointestinal (esophageal and gastric) cancers in the Van region of Eastern Turkey. METHODOLOGY: The patients were diagnosed by esophagogastroscopy, biopsy and histopathological analysis. The control group consisted of 73 healthy subjects. Epidemiological characteristics (age, sex, dietary habits, educational status, life style) were evaluated by questionnaires. Helicobacter pylori infection was diagnosed in nontumoral gastric mucosal biopsy specimens by Giemsa staining. RESULTS: 298 esophageal Ca and 384 gastric Ca patients were diagnosed in seven years (1994-2001). Most of the patients (90%) were of rural origin. Endoscopic prevalence rate of upper gastrointestinal (UGI) Ca was the highest (1 Ca per 7 esophagogastroscopy) in the Van region compared to other parts of Turkey. Esophageal Ca were 1.5 times more common in females whereas gastric Ca were 1.6 times more common in males. Ninety per cent of esophageal cancers were epidermoid and the remaining 10% were adenocarcinomas. Ninety-seven per cent of gastric Ca were adenocarcinomas. Helicobacter pylori infection was not significantly increased in gastric Ca patients compared to controls. CONCLUSIONS: Low educational and socioeconomic status, consumption of smoked, salted, hot, fatty foods, overdrinking hot tea and well water, cigarette smoking, poor intake of fresh fruits and vegetables and poor hygienic conditions were probable culprit factors. PMID- 15816466 TI - Results from the application of extramucous valvular pyloroplasty. AB - BACKGROUND/AIMS: A method for pyloroplasty with complete reconstruction of the pyloric area is presented. METHODOLOGY: The method had been preliminarily tested in dogs and then clinically applied in 28 peptic ulcer patients. The task was solved by pneumatic preparation and ring-shaped removal of pylorus musculature within a zone of 3.0-3.5 cm the integrity of the underlying mucous muff was preserved and the latter was invaginated into the lumen. Thus a circular mucous submucous valve in the region of the gastroduodenal ligament was formed. Due to the preserved anatomical integrity, innervation and blood supply of this mucous submucous layer after its pleating created a zone wide like a normal pylorus between the stomach and duodenum. RESULTS: The duration of the postoperative follow-up was between 6 months and 10 years. The newly created valve looked like a normal pylorus. It was 6 mm thick and 11 mm high and protruded into the lumen. The submucous layer was doubled, richly vascularized, and the muscular layers were continuous. CONCLUSIONS: This operative technique could successfully be applied in gastric surgery for preventing the dumping syndrome and gastric reflux when pyloroplasty is required. PMID- 15816467 TI - Effects of perioperative cimetidine administration on peripheral blood lymphocytes and tumor infiltrating lymphocytes in patients with gastrointestinal cancer: results of a randomized controlled clinical trial. AB - BACKGROUND/AIMS: Cimetidine (CIM) seems to have positive effects on the immune systems of cancer patients. This study was conducted to investigate the effects of perioperative administration of CIM on the peripheral blood lymphocytes, natural killer (NK) cells and tumor infiltrating lymphocytes (TIL) in patients with gastrointestinal (GI) cancer. METHODOLOGY: Forty-nine GI cancer patients were randomized into a treatment group which took CIM in the perioperative period, and a control group which did not take the drug. The treatment was initiated 7 days (d) before operation and continued until 10 d after surgery. At baseline examination, before operation, on the 2nd and the 10th postoperative d, peripheral blood T lymphocytes, helper T cells, T suppressor cells, and NK cells were measured by immunocytochemical method. The surgical specimens were examined for TIL response, and immunohistochemical study was performed to measure the proportion of T and B lymphocytes in the TIL population. RESULTS: In comparison with normal controls, both the treatment and the control groups had decreased T cells, helper T cells and NK cells at baseline. In the control group, total T cells, helper T cells and NK cells declined progressively with the disease course and the decreases became more profound after operation. From the baseline to the 2nd postoperative d, the proportion of total T cells, helper T cells, and NK cells went down from 60.5+/-4.6 to 56.2+/-3.8 percent, from 33.4+/-3.7 to 28.1+/ 3.4 percent, and from 15.0+/-2.8 to 14.2+/-2.2 percent, respectively. On the other hand, there were significant improvements in these parameters after CIM treatment. On the 10th postoperative d, the treatment group had significantly higher percentages of total T cells, helper T cells and NK cells than control group. Moreover, CIM treatment also boosted the TIL response, as was reflected by findings that 68% (17/25) of the patients in the treatment group had significant TIL responses and only 25% (6/24) of the cases had discernible TIL response. CONCLUSIONS: Perioperative application of CIM to GI cancer patients could help restore the diminished cellular immunity boost TIL responses to tumor. PMID- 15816468 TI - Gender and disease activity influence health-related quality of life in inflammatory bowel diseases. AB - BACKGROUND/AIMS: Measurement of health related quality of life (HRQoL) is a new tool to evaluate patients with inflammatory bowel disease (IBD). The aims of this study were to verify reliability and responsiveness of a disease-specific questionnaire [Italian Questionnaire on Quality of Life (IQQoL)], and to assess the relationship between clinical and demographic variables and HRQoL in IBD patients. METHODOLOGY: The IQQoL was submitted to all IBD patients consecutively seen at eight participating Hospitals, and re-administered at follow-up visits. The IQQoL covers intestinal and systemic symptoms, emotional and social function. The higher the score, the worse the HRQoL. RESULTS: 249 patients were enrolled, 106 with Crohn's disease (CD) and 143 with ulcerative colitis (UC). IQQoL was re administered to 134 patients: 98 with unchanged, 17 with worsened and 19 with improved disease activity. The IQQoL was stable over time in patients with stable clinical conditions, and very responsive to change both in patients with improved and worsened disease activity. HRQoL was inversely correlated with disease activity, both in CD and UC. Perception of HRQoL was significantly worse in women than in men. CONCLUSIONS: The IQQoL is a reliable and responsive instrument to assess HRQoL in IBD patients. Active disease is related to poor HRQoL perception. In CD, women, mainly if young, have a worse HRQoL perception than men. PMID- 15816469 TI - A case of Castleman's disease arising from the lesser omentum. AB - A 29-year-old female presented with upper abdominal pain. An upper gastrointestinal radiograph and endoscopy revealed an extra compression in the lesser curvature of the body of the stomach. A computed tomography scan and magnetic resonance imaging revealed a tumor located between the left lobe of the liver and the lesser omentum of the stomach. F-18 fluorodeoxyglucose positron emission tomography revealed high uptake at the tumor in the upper abdomen. In an angiogram, a large hypervascular mass had a prominent vascular supply from the left gastric artery; venous pooling and an enlarged feeding vessel were also apparent. From these results, we suspected that the patient had Castleman's disease arising from the lesser omentum. The patient underwent hand-assisted laparoscopic tumor resection. The resected tumor was an encapsulated mass, the surface of which was smooth and the dimensions of which were 77 x 51 x 43 mm. Based on microscopic findings, we diagnosed hyaline vascular type Castleman's disease. Since surgical intervention, the patient has remained asymptomatic, with no pathologic clinical or laboratory findings. Castleman's disease that occurs in the lesser omentum is extremely rare, and the preoperative diagnosis is very difficult. For the localized type of Castleman's disease, clinical findings are usually improved by complete surgical resection. PMID- 15816470 TI - Carcinoma, a fibrolamellar variant--immunohistochemical analysis of 4 cases. AB - BACKGROUND/AIMS: To analyze, by means of immunocytochemistry, the cases of fibrolamellar variant of hepatocellular carcinoma (FLC), diagnosed in our Department. METHODOLOGY: The material comprised 4 FLC cases of tumors resected surgically. Besides the routine morphological assessment, we used a panel of immunohistochemical stainings including: hepatocellular cytokeratin, CK7, CK19, Ki67, PCNA, chromogranin A, synaptophysin, NSE, insulin, calcitonin, parathormon, CD34, EBV (LMP), Bcl2, cyclin D1. RESULTS: In 3 out of 4 cases, we observed co expression of CK7 with hepatocellular CK. In addition, there was positive staining with some endocrine markers in the majority of patients. In one case, we found strong cyclin D1 immunoreactivity which correlated with EBV (LMP) immunoreactivity, in the same patient. The score of PCNA positivity varied between 15 and 90%. In all cases Ki67 was negative. CONCLUSIONS: The incidence of FLC, among all hepatocellular carcinomas diagnosed in our Department was 5.1%. In accordance with other reports, all our FLC cases were young patients without underlying liver disease. We were unable to find a correlation between FLC cellular immunophenotype, and histological and clinical markers of malignancy. In addition, it appears that PCNA is a better marker of cell-proliferation in FLC than Ki67. The significance of EBV infection in FLC requires further study. PMID- 15816471 TI - Perioperative blood transfusion and survival following curative hepatic resection for hepatocellular carcinoma. AB - BACKGROUND/AIMS: The association between transfusion and recurrence after resection for hepatocellular carcinoma (HCC) is still under debate. The influences of perioperative blood transfusion on survival and recurrence after curative hepatic resection for HCC and prognostic factors in patients with blood transfusion were evaluated. METHODOLOGY: Curative hepatectomy was performed in 210 patients (57%) with and 158 (43%) without perioperative blood transfusion. Prognostic factors were evaluated by univariate and multivariate analysis using Cox's proportional hazards model. RESULTS: Multivariate analysis revealed that perioperative blood transfusion was an independent predictor for recurrence in patients with serum low albumin level (< 3.5 g/dL). In transfused group, stage IV, large tumor size (> or = 5 cm), high value of ICGR15 (> or = 20%), and old age (> or = 60 year) were independent factors of poor disease-free survival. CONCLUSIONS: Perioperative blood transfusion promotes the recurrence of HCC after hepatic resection in patients with hypo-albuminemia. In transfused patients, establishment of strategy for recurrence based on pTNM staging, tumor size, ICGR15, and age may be required to improve survival. PMID- 15816472 TI - Oxidative stress in chronic hepatitis C: a preliminary study on the protective effects of antioxidant flavonoids. AB - BACKGROUND/AIMS: Oxidative stress is involved in chronic hepatitis C, and efforts have been made to influence the disease process with antioxidants. The present study evaluates the protective effects of a phenol-rich processed grain food with superoxide-scavenging properties (trade name antioxidant biofactor AOB). METHODOLOGY: Thirty patients participated in this placebo-controlled double-blind pilot study. AOB was taken orally by fifteen patients for 3 mo at the recommended daily dose of 3x2 sachets, containing 3 g of powder each. Another fifteen patients received a herbal extract with practically no superoxide scavenging properties as a placebo. Oxidative stress biomarkers, aminotransferase levels and viral load were evaluated immediately before and after treatment. RESULTS: AOB treatment considerably improved the antioxidant defenses. Also ALT and AST decreased in 11 of the 15 patients (-11% to -65%, mean -22%, p<0.05). The effects of placebo were not significant. Viral load remained unchanged. Control biopsies were not done after the short interval of 3 mo. There were no adverse effects. After the 3-mo treatment with AOB or placebo, 16 of the 30 patients received conventional antiviral treatment (pegylated interferon alpha and ribavirin). A sustained response was observed in 5 of 9 AOB pretreated patients six mo after discontinuation of the 12-mo antiviral therapy. The 7 patients pretreated with placebo were all non-responders. CONCLUSIONS: These preliminary results are encouraging to conduct more extensive clinical studies combining antioxidant with antiviral treatment in hepatitis C. PMID- 15816473 TI - Evaluation of functional liver reserve in patients with obstructive jaundice undergoing hepatectomy. AB - BACKGROUND/AIMS: In hepatectomy in patients with obstructive jaundice, correct evaluation of the functional liver reserve is sometimes difficult. This study examined whether the preoperative level of total bilirubin in the bile from a predicted remnant liver can be used as a predictor for safe hepatectomy in such patients. METHODOLOGY: Twenty consecutive patients with jaundice who underwent major hepatectomy and in whom all bile from the predicted remnant liver could be collected were included in this study. The level of total bilirubin in the bile was calculated by multiplying bile volume and density obtained by percutaneous transhepatic biliary drainage. Patients were divided into two groups, with (group 1) and without postoperative death due to liver failure (group 2). RESULTS: The level of total bilirubin in the bile from the predicted remnant liver was lower in group 1 than in group 2. Patients did not die of postoperative liver failure if the level of total bilirubin in the bile was maintained, even if the bile volume or density were low. CONCLUSIONS: The preoperative level of total bilirubin in the bile from the predicted remnant liver can be used as a predictor for safe hepatectomy in patients with obstructive jaundice. PMID- 15816474 TI - Progressive hepatocytic fatty infiltration in rats with prehepatic portal hypertension. AB - BACKGROUND/AIMS: Homogenous evolution, with a narrow range of portal hypertension, degree of portosystemic shunt and hepatic atrophy has been described in the experimental model of prehepatic portal hypertension in the rat. However the great differences observed in the rats' liver weight could be attributed to a pathological alteration of the liver. Based on this, we performed an evolutive histological study of the liver. This study shows the existence of a progressive hepatocytic fatty infiltration. METHODOLOGY: Male Wistar rats with portal hypertension induced by triple stenosing ligation of the portal vein at 1 month (group II, n=4) and at 1 year (group IV, n=10) of postoperative evolution were used. The portal pressure, body, liver and splenic weights, types of collateral circulation and degree of mesenteric venous congestion were studied. The intracytoplasmatic lipid microvacuoles were quantified in hepatocytes with an image analyzer (software MIP/CID, Spain). The results were compared with those obtained in control rats with the same evolutive periods (Groups I and III). RESULTS: The hepatic fatty infiltration in Group II (TPVS 1 month) (30.12+/-53.92 micron2) is similar to that presented by Group III (Control 1 year) (16.52+/ 45.20 micron2), while there is an increase (p<0.001) in Group IV (triple portal vein stenosis 1 year) (182.03+/-371.42 micron2) in relation to the other groups studied. The progressive hepatic fatty infiltration in triple portal vein stenosis rats is associated with a decrease of portal pressure and of the incidence of liver hepatic atrophy, portosystemic collateral circulation and mesenteric venous congestion. CONCLUSIONS: TPVS produces progressive hepatocytic fatty infiltration in the rat so that this prehepatic portal hypertension experimental model could also be considered as a hepatic steatosis model. PMID- 15816475 TI - Up-regulation of IL-18 by interferon alpha-2b/ribavirin combination therapy induces an anti-viral effect in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: Recent large prospective trials demonstrated that the combination therapy of interferon (IFN)-alpha/ribavirin significantly increased a sustained virological response rate in patients with chronic hepatitis C. However, the potential mechanism of ribavirin is not clear. METHODOLOGY: Serum interleukin (IL)-18 and HCV-RNA titer were determined before and 2 weeks after administration in patients with chronic hepatitis C, who were treated with ribavirin in combination with IFN-alpha2b (combination group), and with IFN alpha2b alone (monotherapy group). RESULTS: All HCV patients were genotype 1b. In the combination group, the decline of HCV-RNA level by treatment highly correlated with the IL-18 ratio (serum IL-18 level 2 weeks after administration/serum IL-18 level before administration). Similarly, the HCV-RNA level 2 weeks after administration inversely correlated with the IL-18 ratio. In contrast, the IL-18 ratio in the monotherapy group was lower. Furthermore, the decline of HCV-RNA level did not correlate with the IL-18 ratio in the monotherapy group. CONCLUSIONS: This study suggests that ribavirin may contribute to the antiviral effect through up-regulation of IL-18 in combination with IFN in patients with chronic hepatitis C. PMID- 15816476 TI - Risk factors of long-term survival and recurrence after curative resection of hepatocellular carcinoma. AB - BACKGROUND/AIMS: The independent risk factors contributing to long-term survival (> or = 10-year survival rate) and recurrence after curative hepatic resection for hepatocellular carcinoma (HCC) were evaluated. METHODOLOGY: The prognoses were retrospectively analyzed in 247 consecutive patients (187 men and 60 women) treated with curative hepatic resection for HCC and discharged from the hospital. Prognostic factors were evaluated by multivariate analysis using Cox's proportional hazards model. RESULTS: Multivariate analysis revealed that pTNM stage IV, indocyanine green retention rate at 15 minutes (ICGR15) of > or = 20%, tumor size of > or = 5 cm, and positive hepatitis B surface antigen were independent risk factors of overall survival. Stage IV and ICGR15 of > or = 20% were also independent risk factors of disease-free survival. CONCLUSIONS: pTNM stage and ICGR15 may be simple and useful predictors to improve long-term survival and recurrence after curative hepatic resection for HCC. PMID- 15816477 TI - Cirrhosis-related intrathoracic disease. Imaging features in 1038 patients. AB - BACKGROUND/AIMS: To describe imaging features of cirrhosis-related intrathoracic disease. METHODOLOGY: Chest CTs of 1038 cirrhotic patients (mean age 53 yrs; range, 17-79) were evaluated for: bronchoarterial ratio (BAR), arteriovenous malformations, interstitial opacities, emphysema, and pleural effusions. Lymphangiography, pulmonary angiography, cardiac ultrasound and scintigraphy were selectively performed. RESULTS: Mean BAR was 0.83+/-0.19. In two patients with hepatopulmonary syndrome (HPS), mean BAR was 0.55. HRCT detected interstitial lung opacities in 15 patients. Signs of fibrosis were seen in 7 (only two associated to biliary cirrhosis) and interstitial edema in 8. Accurate pattern recognition was achieved in 10/15 cases (66.6%). Of the 93 patients with emphysema only one had documented alpha1-AT deficiency (1.08%). Multiple type 1 vascular dilatations were visualized in two patients with HPS. Hepatic hydrothorax was present in 49 patients (4.72%); right-sided in 34 (69.4%), bilateral in 9 (18.4%) and left-sided in 6 (12.2%). Hepatic chylothorax was confirmed in 3 patients. Lymphangiography demonstrated the site of leakage and the engorged thoracic duct. CONCLUSIONS: CT can identify intrathoracic pathology associated with liver disease. Decreased BAR is highly specific for HPS. However, a multimodality approach is necessary to depict cases of liver origin. PMID- 15816478 TI - Additional effect of low iron diet on iron reduction therapy by phlebotomy for chronic hepatitis C. AB - BACKGROUND/AIMS: Iron-induced oxidative stress plays an important role in the pathogenesis of chronic hepatitis C. Both phlebotomy for removing body iron stores and low iron diet for minimizing portal iron supply to the liver have been shown to improve serum transaminase levels in patients with the disease. However, the cooperative effects of phlebotomy and low iron diet have not yet been elucidated in detail. METHODOLOGY: A pilot study was undertaken to investigate whether a low iron diet could improve the efficacy of phlebotomy in iron reduction therapy. Of 21 patients diagnosed with chronic hepatitis C, 10 patients were treated with phlebotomy alone (group A) while 11 patients were treated with a low iron plus phlebotomy (group B). Phlebotomy was repeated biweekly until serum ferritin levels reached 10 ng/mL in both A and B groups. In addition, a low iron diet (iron intake of 8 mg/day or less) was recommended for group B, followed by estimation of iron intake from daily diet records. RESULTS: Serum alanine aminotransferase levels were significantly improved from 106+/-30 to 68+/-22 IU/L (p<0.005, paired t-test) in group A and from 100+/-33 to 46+/-10 IU/L (p<0.002, paired t-test) in group B. The enzyme levels after treatment were significantly higher in group A (p<0.02, non-paired t-test), which showed a higher upward distribution of the enzyme activity. The estimated dietary iron intake in group B was reduced from 17.6+/-6.1 to 8.2+/-3.7 mg/day. CONCLUSIONS: These findings suggest that phlebotomy alone does not completely remove iron-induced oxidative stress and a low iron diet induces an additional effect in iron reduction therapy for chronic hepatitis C. PMID- 15816479 TI - Growth factor NUN increased AKR1C2 expression by activated CDK2 related RB signal transduction pathway in human liver cancer cell line. AB - BACKGROUND/AIMS: The investigation is to understand whether polypeptide growth factor NUN can enhance AKR1C2 transcriptional expression in human SMMC7721 liver cancer cell line or not. METHODOLOGY: SMMC7721 cell was starved in serum-free medium overnight before NUN treatment, and cells were then incubated with NUN for different times and dosage. These cells lysates were prepared to examine AKR1C2 expression by Western blot. Total RNA was extracted from these cells to analyze the change of AKR1C2 mRNA by Northern blot. RESULTS: NUN increased expression of AKR1C2 in SMMC7721 liver cancer cell in a time- and dosage-dependent manner and this increase resulted from up-regulation of AKR1C2 mRNA expression. The inhibitor of CDK2 inhibited the up-regulation of AKR1C2 by NUN. CONCLUSIONS: These results showed that NUN can enhance AKR1C2 transcription by activated CDK2 related RB signaling pathway, leading to increased AKR1C2 expression in SMMC7721 liver cancer cell. PMID- 15816480 TI - The influence of radiofrequency ablation on hepatic vessels in porcine liver. AB - BACKGROUND/AIMS: The objective of this study was to clarify the influence of radiofrequency ablation on the portal and hepatic vein. METHODOLOGY: Hepatic radiofrequency ablation was performed on 18 swine (mean weight, 22 kg). The livers were removed immediately, 1 week or 3 weeks following ablation. Vessel patency and diameter were determined by computed tomography, and the severity of endothelial injuries was determined histopathologically. RESULTS: The patency rate for portal and hepatic veins with diameters less than 3.0 mm decreased sequentially. At three weeks following ablation, the patency rate for vessels with diameters more than 3.0 mm was significantly higher (P<0.05) compared to those with diameters less than 2.0 mm. In portal and hepatic veins contiguous to ablated hepatic parenchyma, the endothelium with intimal thickening was found at 1 to 3 weeks following ablation, although its detection rate was decreased until 3 weeks. CONCLUSIONS: The portal and hepatic vein with diameters more than 3.0 mm in radiofrequency lesions maintained high patency. The endothelium seems to play an important role for maintaining vessels patency and the distal hepatic tissue following ablation. We believe that this maintained vascular patency may be the advantage of radiofrequency ablation over the PEIT. PMID- 15816481 TI - The concentration of Fas in the serum and liver tissue in viral chronic hepatitis B patients treated with lamivudine. AB - BACKGROUND/AIMS: HBV infection stimulates cellular synthesis and Fas expression. The dependence of the Fas concentration in the liver tissue and serum was estimated in reference to the elimination of HBV-DNA, necroinflammatory changes and fibrotic activity in chronic hepatitis B patients treated with lamivudine. METHODOLOGY: Included in the study were 31 patients with chronic hepatitis B treated with lamivudine for 12 months. Fas in the liver tissue was measured before therapy. Fas and beta-2M in the serum, CD4 lymphocytes in the blood were also measured before therapy and after 1 and 3 months of treatment. The concentration of Fas in the liver tissue of patients with HBV infection was compared to concentration of Fas in the liver tissue of patients with alcoholic liver disease. Fas in the liver tissue and serum was measured with use of the ELISA test, firm Bender Med Systems (Austria). RESULTS: At the end of treatment, 71% (23/31) patients ALT had normalized, 45% (14/31) eliminated HBV-DNA and 39% (12/31) eliminated antigen HBe. The concentration of Fas was higher in the liver tissue of chronic hepatitis B patients (1009 ng/g) in comparison to the patients with alcoholic disease of the liver (631 ng/g). The activity of fibrosis in the liver tissue did not cause any differences in the Fas concentrations. The concentration of Fas in the liver tissue was not dependent on the effectiveness of the antiviral therapy. No correlation between the concentration of Fas in the liver tissue in comparison to the serum was observed (Pearson=0.169). The highest concentration of Fas in the serum was observed in patients who did not eliminate HBV-DNA (30.7 pg/mL) and in patients with moderate necroinflammatory changes (30.2 pg/mL). Among the patients, who eliminated HBV-DNA, the concentration of Fas in the serum normalized after the third month of treatment (18.2 pg/mL). In patients with a poor response to the therapy the concentration of Fas in the serum remained above the preferred values (25.2 pg/mL). CONCLUSIONS: The concentration of Fas in the serum did not correlate with the concentration of Fas in the liver tissue of chronic hepatitis B patients. Allowing concentration of Fas before therapy and fast normalization this protein in the serum could be a good prognostic indicator of treatment. PMID- 15816482 TI - Risk factors of intrahepatic recurrence after curative resection of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Although the risk factors for the development of intrahepatic recurrence after hepatectomy for hepatocellular carcinoma (HCC) have been widely studied, little attention has been given to the prognostic factors affecting such patients. METHODOLOGY: Intrahepatic recurrence occurred in 105 (56%) of 188 patients who underwent curative hepatic resection of HCC and were discharged from the hospital. Among them, 17 (16%) also had simultaneous extrahepatic recurrence. Independent prognostic factors were evaluated by multivariate analysis using Cox's proportional hazards model. RESULTS: Multivariate analysis revealed that presence of extrahepatic recurrence, hepatitis B, and non-surgical treatments for recurrence were independent predictors of poor overall survival after initial hepatic resection or after recurrence. Risk factors of extrahepatic recurrence were young age, solitary and large HCC, high hepatitis activity, and large amount of intraoperative blood loss and blood transfusion. CONCLUSIONS: Survival of patients with intrahepatic recurrent HCC after resection should be stratified by the type of recurrence, type of hepatitis, and type of treatment for recurrence. PMID- 15816483 TI - Postoperative complications of hepatectomy for hepatocellular carcinoma in relation to liver fibrosis. AB - BACKGROUND/AIMS: Hepatectomy for hepatocellular carcinoma remains associated with significant morbidity and mortality, despite improved surgical techniques and perioperative management. The relationship between the extent of liver fibrosis and postoperative complications is controversial. METHODOLOGY: We studied 50 patients in whom hepatectomy for Child-Pugh grade A or B had been indicated. Surgical procedures were determined by the indocyanine green retention rate after 15 minutes (ICGR15). The extent of liver fibrosis was graded according to the Desmet classification. The frequency and grade of severity of postoperative complications were evaluated. RESULTS: Postoperative complications occurred in 24 patients (48%), the frequency being significantly higher among those with liver cirrhosis. However, no significant difference in the incidence of major complications was observed between cirrhotic and non-cirrhotic patients (p=0.311). There was no association between grade of severity of postoperative complications and extent of fibrosis. CONCLUSIONS: When the surgical indication and procedure were selected based on the Child-Pugh classification and ICGR15 value, respectively, the extent of liver fibrosis did not influence the incidence of major postoperative complications. In particular, there was no significant difference in the incidence of life-threatening postoperative complications between cirrhotic and non-cirrhotic patients. PMID- 15816484 TI - Recurrence of primary hepatocellular carcinoma after hepatectomy--differences related to underlying hepatitis virus species. AB - BACKGROUND/AIMS: Studies on post-hepatectomy prognoses by infecting viral species have only been rarely reported. METHODOLOGY: The patients who had undergone hepatectomy for primary hepatocellular carcinoma (HCC) over the past 10 years at our hospital were divided into three groups based on their underlying causal diseases: hepatitis B (B type), hepatitis C (C type), and non-viral hepatitis including alcoholic hepatitis, and their backgrounds and long-term results after hepatectomy were comparatively investigated. RESULTS: As for tumor factors, the tumor diameter was 3.7+/-2.1 cm in C type patients, which was significantly smaller than the 5.2+/-3.3 cm diameter in B type patients (p=0.01) and 7.9+/-5.6 cm in non-viral patients (p=0.001). However, the frequencies of intrahepatic metastasis, portal vein infiltration, capsule formation and capsule infiltration did not differ significantly among the three groups. Although there was no significant difference observed among the three groups for the recurrence-free survival time, HCC had recurred at as early a stage as within 1 postoperative year in 50% of non-viral patients. CONCLUSIONS: The size of a tumor upon hepatectomy and the presence or absence of intrahepatic metastasis were found to be significant factors contributing to the postoperative recurrence of HCC. It is possible that the recurrence-free postoperative survival time may be prolonged by earlier detection of HCC, particularly for the B type and the non-viral type in which the tumor size was already large at the time of surgery. PMID- 15816486 TI - Long-term survival of intrahepatic cholangiocarcinoma with hilar lymph node metastasis and portal vein involvement. AB - We report the case of a very rare 6-year disease-free survivor of intrahepatic cholangiocarcinoma with hilar lymph node metastasis and portal vein involvement. A 76-year-old female with liver dysfunction was referred to our institution. Contrast-enhanced computed tomography showed a 5-cm low-density tumor with irregular marginal enhancement in the left and caudate lobes of the liver. Cholangiography revealed complete obstruction of the left hepatic bile duct. Angiography showed obstruction of the left branch of the portal vein. Metastasis to the hilar lymph nodes was disclosed at surgery. The patient underwent left hepatectomy with caudate lobectomy, resection of the extrahepatic bile duct, and lymphadenectomy. The total vascular exclusion of the liver was used for hepatectomy and reconstruction of the portal vein. Microscopically, the tumor was a poorly differentiated adenocarcinoma with many infiltrating lymphocytes, and extensive necrosis was present within the tumor. The experience gained in the present case suggests that aggressive surgery may be a potential approach to provide a hope of long-term survival for patients with intrahepatic cholangiocarcinoma despite the presence of regional lymph node metastasis and vascular invasion. PMID- 15816485 TI - Ursodeoxycholic acid (UDCA) suppresses liver interleukin 2 mRNA in the cholangitis model. AB - BACKGROUND/AIMS: Ursodeoxycholic acid (UDCA) is used globally as the drug of first choice for the treatment of primary biliary cirrhosis (PBC). The mechanism by which UDCA exerts its effect has been clarified mainly by in vitro studies. However, no other studies have so far been successful in defining the expression profiles of relevant cytokines in experimental PBC models. METHODOLOGY: In this study, we established an immune-mediated cholangitis mouse model by immunizing mice with an intraperitoneal injection of carbonic anhydrase (CA)-II every other week, for a total of three injections. After the administration of UDCA, the animals were examined for the hepatic histopathology and liver enzyme levels in the serum, as well as the cytokine mRNA contents in the liver. RESULTS: After the administration of UDCA, peribiliary cell invasion decreased, but the change of hepatic enzyme was not observed. The quantities of interleukin (IL)-2 mRNA in the liver were all elevated in the CA-II group as compared with the control group in a semiquantitative assay. The quantities of IL-2 mRNA were significantly decreased in the CA-II+UDCA group compared with the CA-II group. UDCA suppressed the production of IL-2 and had the tendency to suppress the production of IL-4, and the suppression of IL-2 was predominant, compared to the suppression of IL-4, but UDCA did not significantly effect the expression of interferon (IFN)-gamma mRNA, IL-6 mRNA, IL-10 mRNA. CONCLUSIONS: UDCA predominantly suppressed IL-2 mRNA compared to IL-4 mRNA in the liver of the cholangitis model. The results partially clarified the mechanism by which UDCA exerts its effect on PBC. PMID- 15816487 TI - Caroli's disease. Report of 5 cases and review of literature. AB - In 1958 Caroli was the first to define exactly in literature the congenital dilatations of the intrahepatic bile ducts, concerning a segment, a lobe, or both lobes of the Liver. Later Todani involved it in his classification of the bile duct cysts as type V. Caroli's disease (CD) is quite a rare entity (about 150 cases reported in literature). In CD, stenosis and dilatation cause initially biliary stasis, leading to intrahepatic biliary lithiasis and secondary infection. The spontaneous course of CD is dominated by biliary infection: cholangitis (cholangiohepatitis), septicemia and intrahepatic and subphrenic abscesses. That is why the disease must be recognized before serious complications develop. The authors report about 5 cases of Caroli's disease--2 women and 3 men. Diffuse spread in one patient (25-year-old man) and in four- left lobe involved predominantly. Most common signs and symptoms were: fever, cholangitis, upper quadrant abdominal pain, hepatosplenomegaly, obstructive jaundice, Charcot's triad (in two). Importance for the exact diagnosis involves the use of ultrasonography (US), computer tomography (CT) scan, endoscopic cholangiopancreatography (ERCP), intraoperative cholangiography and cholangioscopy, allowing early diagnosis and therefore a better therapeutic and surgical approach. PMID- 15816488 TI - Living donor liver transplantation in a recipient with anti-Fy(a) and Jk(a) antibodies. AB - We report a case of a living donor liver transplantation using right lobe graft to a recipient having anti-Fy(a) and Jk(a) antibodies. The red blood cell (RBC) antigens of the donor were Fy (a+) and Jk (a-). We attempted to eliminate donor RBCs remaining in the graft by perfusing histidine-tryptophan-ketoglutarate. Further, Fy (a-)/Jk (a-) RBC concentrates were transfused during the operation. However, the anti-Fy(a) titer increased approximately 8-fold on the seventh postoperative day. On the same day, serum levels of transaminase and total bilirubin increased presumably due to acute cellular rejection. Steroid administration immediately reduced levels of transaminase, total bilirubin and anti-Fy(a) titer. The increase of anti-Fy(a) titer may be due to a secondary immune response to the donor's Fy(a) antigen on RBCs remaining in the graft. PMID- 15816489 TI - The tree shrews: useful animal models for the viral hepatitis and hepatocellular carcinoma. AB - Hepatitis B virus (HBV)-induced hepatitis and hepatocellular carcinoma (HCC) are major diseases worldwide. HBV infection and chemical carcinogens such as aflatoxin B1 are known to be two key factors in the development of HCC. Animal models for hepatitis and HCC are very useful in the in vivo studies of mechanism involved in the development and prevention of these diseases and the pre-clinical research of drugs for the treatment of these diseases. Now, several animals, such as woodchucks, ground squirrels, chimpanzees, ducks and tree shrews, have been used to establish hepatitis and HCC models. HCC occurs in some woodchucks and ground squirrels that are infected with their own hepatitis viruses and exposed to carcinogens. Chimpanzees and ducks can be infected with human and duck hepatitis B viruses, respectively, but HCC is rarely observed in these animals. The tree shrews are non-rodent, small animals and close to primates in evolution. This review focuses on the establishment of human HBV-induced hepatitis and human HBV-associated HCC in tree shrews and their applications in the study of HCC development. PMID- 15816490 TI - Pancreaticoduodenectomy with closing the pancreatic stump vs. standard Whipple's procedure: a non-anastomotic technique. AB - BACKGROUND/AIMS: Pancreaticoduodenectomy is commonly used for the surgical treatment of malignancies of the ampulla of Vater, duodenum, head of pancreas, and distal common bile duct (CBD). Pancreatic fistula and anastomotic leakage are the common fatal complications of the procedure. Management of the remaining stump is the most important part of pancreaticoduodenectomy in preventing fistula and leakage. We describe a non-anastomotic procedure that has fewer complications. METHODOLOGY: Wirsung's duct was ligated with interrupted sutures after pancreaticoduodenectomy. Cut edge of pancreatic stump was then sutured. Drainage of the stump field was performed with a Petzer drain. RESULTS: Of 6 patients who were studied 3 were men and 3 were woman. The mean age was 59.19 years. There was no pancreatic fistula, anastomotic leakage, significant weight loss, far elevation in serum amylase, pancreatitis and oral intake serious problems within the follow-up months (median=7 months, min=2 months, max=20 months). CONCLUSIONS: Non-anastomotic options such as this necessitate the use of pancreatic enzyme supplementation, but low rate of complications and simplicity of the procedure make it an operation of choice. PMID- 15816491 TI - Laparoscopic distal pancreatectomy for neuroendocrine tumors of the pancreas. AB - Although neuroendocrine tumors of the pancreas are traditionally managed by laparotomy, these rare neoplasms may be amenable to laparoscopic surgical resection. We present our experience with laparoscopic distal pancreatectomy in two such patients, and discuss the operative technique with emphasis on organ preservation. Two female patients aged 63 and 69 years presented with clinical and biochemical features of an insulinoma and a vasoactive intestinal peptide secreting tumor (VIPoma), and were found on cross-sectional imaging to have 1.2 cm and 4.5-cm solitary tumors in the tail of the pancreas. They underwent laparoscopic distal pancreatectomy with and without preservation of splenic vessels and spleen respectively. Both procedures were completed laparoscopically. The operating time was 180 and 210 minutes respectively. There were no postoperative complications. The postoperative hospital stay was 4 and 14 days respectively. Histology revealed a benign insulinoma and a malignant VIPoma with lymph node metastases respectively. Laparoscopic distal pancreatectomy for neuroendocrine tumors of the pancreas may be accomplished safely, with preservation of the spleen and splenic vessels in benign disease, and with benefits to the patients in terms of postoperative recovery. PMID- 15816492 TI - Inflammatory myofibroblastic tumor (inflammatory fibrosarcoma) of the pancreas: a case report. AB - Inflammatory myofibroblastic tumors (inflammatory fibrosarcomas) of the pancreas are extremely rare. We report a 29-year-old woman who underwent pancreatoduodenectomy for a 6-cm tumor of the pancreas head causing obstructive jaundice. Tumor involvement was local, without apparent metastasis. The tumor was composed of proliferating fibroblastic/or myofibroblast-like spindle cells and aggregates of chronic inflammatory cells in a fibromyxoid matrix. Immunohistochemical examination demonstrated reactivity only to vimentin. This tumor has often been found in the peritoneal cavity, the retroperitoneum, or the pelvic cavity, but only very rarely in the pancreas. PMID- 15816493 TI - Vascular endothelial growth factor C (VEGF-C) expression in pT2 gastric cancer. AB - BACKGROUND/AIMS: The purpose of this study was to determine whether the vascular endothelial growth factor C (VEGF-C) protein expression was related to the clinicopathologic features of patients with pT2 (primary tumor invasion of muscularis propria or subserosa) gastric cancer. METHODOLOGY: The expression of VEGF-C protein was investigated retrospectively in 102 patients with pT2 gastric cancer. Immunohistochemical staining of the paraffin sections was performed using a polyclonal antibody to VEGF-C. RESULTS: Normal gastric mucosa was not immunoreactive with an anti-VEGF-C antibody. Among the 102 tumors examined, 27 (26.5%) showed high expression of VEGF-C protein. No staining was observed in the normal tissue surrounding the tumor. There were no significant differences in age, gender, or histological types. With regard to the clinicopathological characteristics, significant differences were observed in depth of tumor invasion (muscularis propria vs. subserosa; p<0.05), lymph node metastasis (p<0.001), and stage grouping (p<0.001). The prognosis for VEGF-C-positive patients was worse than that for VEGF-C-negative patients in terms of overall survival, and VEGF-C expression was an independent prognostic indicator (p=0.023) by multivariable analysis. CONCLUSIONS: Determination of VEGF-C expression is important in predicting nodal metastasis and poor clinical outcome in pT2 gastric cancer patients. PMID- 15816494 TI - Inhibitory remnant stomach motility by duodenal distention after B-I gastrectomy. AB - BACKGROUND/AIMS: Although control of gastric motility is a highly regulated process, B-I gastrectomy causes accelerated gastric emptying. We found few reports so far concerned with the mechanical effect of balloon distention of the duodenum on the remnant stomach in B-I dogs. METHODOLOGY: Four weeks after conventional distal gastrectomy with B-I reconstruction procedure, a balloon was inserted into the duodenum, and strain gauge force transducers (SGTs) were sutured onto the wall of the gastric remnant and duodenal serosa in beagles. After a two-week postoperative recovery period, gastro-duodenal motility was measured with SGTs prior to distention, during distention, and after evacuation of the balloon. The motility index (MI) was calculated every 30 minutes. RESULTS: Upon duodenal distention with 4 mL of water, motility of gastric remnant was significantly inhibited, however, no reduction in duodenal MI was observed. The response was maintained for as long as duodenal distension continued. Ninety minutes after evacuation of the balloon, MI returned to predistension levels. Increased plasma levels of cholecystokinin (CCK) were observed upon distention of the duodenal balloon. CONCLUSIONS: Motility of the remnant stomach in B-I dogs was inhibited by duodenal distension. Increased levels of plasma CCK during distention might play a role in this feedback mechanism. PMID- 15816495 TI - Pre- and postoperative electrogastrography in patients with gastric cancer. AB - BACKGROUND/AIMS: To investigate the changes of electrogastrography (EGG) after stomach resection, fasting and postprandial EGG were recorded in 10 volunteers and 23 gastric cancer patients who had undergone total or subtotal gastrectomy. METHODOLOGY: EGG signals in patients were recorded before and after surgery and were analyzed by power spectrum. RESULTS: The power spectrum components at three cycles/minute (cpm) as normal subject, had completely disappeared in all patients after total gastrectomy, and irregular powers, especially under 9 cpm, had increased. These results suggest that the activity of the 3-cpm component is specific for the stomach. In subtotal gastrectomy patients, 3-cpm power peaks were clearly observed in seven of twelve patients, but it was not recorded in the remaining five patients, who showed irregular powers similar to that in patients after total gastrectomy. Therefore, it was speculated that the area containing the "pacemaker" of gastric electric potentials was removed during gastrectomy in these five patients. The postprandial dip, which is considered to reflect autonomic nervous function, was not observed in six of seven subtotal gastrectomy patients who demonstrated the 3-cpm component in their EGG, and it was speculated that the gastric branch of the vagus nerve in these six patients might have been partially removed by surgery. CONCLUSIONS: These findings suggest that EGG is available as a non-invasive method to evaluate the motility and autonomic function of the remnant stomach after gastrectomy. PMID- 15816497 TI - Primary gastric lymphoma: a clinicopathologic study. AB - BACKGROUND/AIMS: This study aims to define the clinicopathologic criteria of primary gastric lymphoma in view of MALT concept and to present the outcome after different treatment modalities. METHODOLOGY: Seventy-six cases of primary gastric lymphoma treated between January 1980 and December 2001 were reviewed. All tissue specimens (endoscopic or surgically resected) were re-examined. Tumors were staged according to Ann Arbor staging system and the Musshoff modification (IE in 30.3%, IIE in 39.4% and IIIE in 30.3%). Sixty patients underwent gastrectomy (partial or total) with postoperative chemotherapy for 32 patients. Sixteen patients were treated by chemotherapy only. The mean follow-up period was 15 years (range, 6 months to 21 years). RESULTS: Primary gastric lymphoma represented 69.1% of cases of gastrointestinal lymphoma and 16.2% of all gastric malignancy. The mean age was 45 years and male to female ratio was 2.3:1. Epigastric pain was the commonest symptom (in 88.2%). Ulcer-like lesions were the commonest (65.8%) and the most commonly involved site was the lower third (48.7%). The resectability rate was 80%. The operative mortality rate was 2.7%. Another 2 cases died after partial gastrectomy and chemotherapy. Four cases in the chemotherapy group (25%) died. Tumor recurrence occurred in 4 cases (out of 32) after gastrectomy and chemotherapy (12.5%), 2 of them died and 2 were cured by chemotherapy. The mean overall survival was 18.49 years, survival was 20.28 years after gastrectomy, 15.48 years after gastrectomy with chemotherapy and 5.76 years after chemotherapy (p=0.0056). CONCLUSIONS: Primary gastric lymphoma is not an uncommon tumor. Gastritis-like lesions are rare. If the tumor is resectable, gastrectomy will provide the most accurate means of diagnosis, staging and locoregional control of the disease. PMID- 15816496 TI - Efficacy of a triple therapy with rabeprazole, amoxicillin, and faropenem as second-line treatment after failure of initial Helicobacter pylori eradication therapy. AB - BACKGROUND/AIMS: Triple therapy consisting of lansoprazole, amoxicillin, and clarithromycin (LAC regimen) is widely used to eradicate Helicobacter pylori in Japan. However, the need for appropriate treatment after failure of initial therapy to eradicate H. pylori has been increasing. We therefore assessed the efficacy of a combination of rabeprazole, amoxicillin, and faropenem for second line eradication therapy. METHODOLOGY: The subjects were 116 patients positive for H. pylori infection. Patients initially received lansoprazole 60 mg/day, amoxicillin 1500 mg/day and clarithromycin 400 mg/day in two divided doses for 7 days. Patients in whom eradication treatment failed were given rabeprazole 20 mg/day and amoxicillin 1500 mg/day in two divided doses, and faropenem 600 mg/day in three divided doses (RAF regimen) for 7 consecutive days. H. pylori status was assessed by the 13C-urea breath test combined with rapid urease test or H. pylori culture method 8 weeks after completion of therapy. Susceptibility to clarithromycin was determined by the agar dilution method, and genetic polymorphism of CYP2C19 was analyzed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The initial H. pylori eradication rate with the LAC regimen was 76.4% (84/110). Assessment of the CYP2C19 genotypes of the patients in whom eradication therapy failed revealed that homozygous extensive metabolizers accounted for 70.0% (16/23) and heterozygous extensive metabolizers for 30.0% (7/23), with no poor metabolizers. The acquired resistance rate for clarithromycin was 52.0% (12/23). The success rate of re-eradication with the RAF regimen was 91.3% (21/23) with no serious adverse effects. CONCLUSIONS: Triple therapy comprising rabeprazole, amoxicillin, and faropenem is effective for second-line eradication treatment of H. pylori infection, regardless of the genetic polymorphism of CYP2C19 or the presence of resistance to clarithromycin. PMID- 15816498 TI - Ets-1 expression in gastric cancer. AB - BACKGROUND/AIMS: Ets-1 protein is a transcription factor that has been implicated in both tumor invasion and neovascularization. The purpose of this study was to investigate the clinical significance of Ets-1 expression in gastric cancer. METHODOLOGY: We examined the expression of Ets-1 protein by immunohistochemical staining in tissues obtained from 124 primary gastric cancers. Ets-1 expression was detected by automated immunodetection (Ventana Medical Systems Inc., Tucon, AZ, USA). RESULTS: Ets-1 was not expressed in normal gastric epithelium. Ets-1 staining was defined as positive in 51.6% (64 of 124) of gastric cancer cases. There were no significant differences in age, gender, or histopathological grading. With regard to clinicopathological characteristics, significant differences were observed in the depth of tumor invasion, lymph node and distant metastases, increased tumor stage, and histological lymphatic and venous invasion. Patients with Ets-1 expression demonstrated poorer survival rates than patients without Ets-1 expression, as determined by the log-rank test (p=0.017), but multivariable analysis showed that the expression of Ets-1 was not an independent prognostic indicator (p=0.370). CONCLUSIONS: Ets-1 expression in gastric cancers is correlated with histological prognostic indicators and is a useful marker for predicting the outcome for patients with gastric cancer. PMID- 15816499 TI - Editorial overview: The 'sartans': is it premature to consider PPARgamma receptor agonism a bonus? PMID- 15816500 TI - Cardiovascular disease in chronic kidney failure: is there a role for vitamin D analogs? AB - Vitamin D3 is modified by vitamin D3-25-hydroxylase in the liver, and 25 hydroxyvitamin D3-1alpha-hydroxylase in the kidney, to form the active metabolite, 1,25-dihydroxyvitamin D3. Chronic kidney disease (CKD) is characterized by reduced synthesis of 1,25-dibydroxyvitamin D3, inadequate renal phosphate clearance and calcium imbalance, secondary hyperparathyroidism (SHPT) and bone disease. CKD patients encounter a much higher risk of cardiovascular disease (CVD) than the general public. The cardiovascular risk factors for CKD patients include conventional factors such as age, gender, hypertension, diabetes, dyslipidemia and smoking, and non-conventional factors, such as anemia, uremia, reduced vascular compliance, inflammation and various hormonal factors. Several vitamin D analogs are currently available for the treatment of SHPT, and recent clinical data show that these analogs provide survival benefit for CKD patients in the order of paricalcitol > calcitriol > no vitamin D analog, independent of parathyroid hormone and calcium. Moreover, the survival benefit seems to be associated with cardiovascular causes. The observations made from these clinical studies raised intriguing questions about the involvement of the vitamin D receptor locus (VDR) in the cardiovascular system. This review discusses recent data regarding the role of vitamin D and its analogs in the CVD associated with CKD. PMID- 15816501 TI - Therapeutic targets for prevention and regression of progressive fibrosing renal diseases. AB - Renal fibrosis complicates most chronic renal diseases, leading to a progressive loss of function and ultimately resulting in terminal renal failure. Molecular mechanisms underlying the development and progression of renal fibrosis have been increasingly identified, and much progress has been made towards a better understanding of the roles of different growth factors/cytokines and regulators of matrix turnover, as well as of the interactions between renal inflammation and fibrosis. This review focuses on recent advances in the identification of novel targets, as well as the development of new therapeutic tools for use in the treatment of progressive fibrosing renal diseases. Using our growing knowledge, treatment strategies need to be identified that prevent progression more effectively, as well as inducing regression of developed renal fibrosis. This is likely by combining compounds that interfere with a variety of targets simultaneously. PMID- 15816502 TI - Epithelial to mesenchymal transition as a triggering factor of peritoneal membrane fibrosis and angiogenesis in peritoneal dialysis patients. AB - Peritoneal fibrosis (or sclerosis) is a complication of peritoneal dialysis (PD) and includes a wide spectrum of peritoneal structural changes, ranging from mild inflammation to severe sclerosing peritonitis and encapsulating-sclerosing peritonitis. In parallel with fibrosis, the peritoneum shows a progressive increase in capillary number (angiogenesis) and vasculopathy, which are involved in both the elevation of small solute transport across the peritoneal membrane and ultrafiltration failure. The most important substances from the PD solutions responsible for peritoneal deterioration are glucose and glucose degradation products, which stimulate transforming growth factor (TGF)beta and vascular endothelial growth factor (VEGF) production by mesothelial cells (MCs). TGFbeta is a potent pro-fibrotic factor and induces epithelial-mesenchymal transition (EMT) of the MC. Local production of VEGF during PD appears to play a central role in the processes leading to peritoneal neo-angiogenesis and functional decline. This review discusses the mechanisms implicated in peritoneal structural alteration and points to EMT of MC as the protagonist and initiator of peritoneal membrane injury, through an increment of the submesothelial fibroblast population. We also propose possible mechanisms of regulation and new targets for inhibition of EMT. PMID- 15816503 TI - Progress in the development of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers. AB - Angiotensin II is a potent vasoconstrictor that stimulates aldosterone secretion. Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) are currently in use for the treatment of hypertension and chronic heart failure. Clinical trials show ACEIs to be effective, but with significant side effects, while ARBs provide a useful, more well-tolerated, alternative. ARBs are based on the structure of losartan, and efficacy has been enhanced by development of longer-lasting insurmountable antagonists. Combination therapy is another option; however, there is also scope for development of new molecules that exploit our increased understanding of angiotensin receptor structure and function. PMID- 15816504 TI - P2Y receptor antagonists in thrombosis. AB - The dual role of P2Y1 and P2Y12 receptors in platelet aggregation by ADP has been firmly established, based on the action of selective inhibitors, gene targeting in mice and human genetic evidence. Both of these receptor subtypes constitute targets for antithrombotic agents, and compounds with a dual action might also be of interest. However, the agents currently on the market (ticlopidine and clopidogrel), or known to be in development (cangrelor, AZD-6140 and prasugrel), all target the P2Y12 receptor. The thienopyridines (ticlopidine, clopidogrel and prasugrel) irreversibly inactivate the P2Y12 receptor via the covalent binding of an active metabolite generated in the liver, while the other compounds are competitive antagonists. Cangrelor, an ATP derivative, is suitable for intravenous perfusion, whereas AZD-6140 is in clinical development as an orally active agent. PMID- 15816505 TI - Novel approaches to using PDE4 inhibitors for antihypertensive therapy. AB - Phosphodiesterase (PDE)4 inhibitors are effective vasodilators. In addition to the treatment of asthma and other inflammatory diseases, such as chronic obstructive pulmonary disease, wide-spectrum PDE4 inhibitors may also be developed for use in antihypertensive therapy, especially in patients with impaired renal function and elevated pulmonary arterial pressure. Rational approaches to overcoming the intrinsic emetogenic response associated with PDE4 inhibition are discussed, and a dual-action drug development that incorporates L type Ca2+ channel antagonism and selective PDE4 inhibition is proposed. The potential for research and development of such a dual-action agent should eventually provide healthcare professionals with a new category of therapeutic options with which to attempt to combat hypertension. PMID- 15816506 TI - Two patterns of ion channelopathy in the myocardium: perspectives for development of anti-arrhythmic agents. AB - Cardiac arrhythmia remains a significant problem, due to the high morbidity and mortality associated with cardiovascular diseases with prominent cardiac remodeling. There is still a lack of effective drugs with which to combat this life-threatening disorder. The abnormal electrophysiological properties of the heart can be explained in terms of ion channels and channelopathy and, in recent years, advances have been made in understanding these properties. There are two patterns of ion channelopathies in the diseased heart: Single insufficiency disorder, which is attributed to mutations in genes, and a multiple derangement of channels. Malignant arrhythmias in a diseased heart usually occur when ventricular hypertrophy is evident, and when they are associated with abnormal repolarization. Abnormalities in the ryanodine receptor-calcium release channel complex (RyR)2, FK-506 binding protein (FKBP 12.6), cardiac sarcoplasmic reticulum calcium ATPase (SERCA2a) and phospholamban (PLB) are involved in the initiation of cardiac arrhythmias, and can be identified as targets for therapeutic interventions. PMID- 15816507 TI - Novel antithrombotic strategies in cardiovascular diseases. AB - Ischemic cardiovascular diseases represent the most common cause of mortality and morbidity in the western world, and atherothrombosis occupies a central role in their pathophysiology. Venous thrombi, which form under low shear conditions, are predominantly composed of fibrin and red cells, while arterial thrombi form under high shear conditions and are composed primarily of platelet aggregates held together by fibrin strands. Several successful strategies targeting specific steps in coagulation and platelet function or interaction have been developed to prevent or treat atherothrombotic disorders. Intense research is currently underway in an effort to develop more safe and effective compounds, such that novel antithrombotics are emerging to target specific steps in the coagulation cascade, as well as in pathways of platelet adhesion, activation and aggregation. This review will focus on the recent advances in research in this fast-evolving field. PMID- 15816508 TI - Plant sterols and their role in combined use with statins for lipid lowering. AB - Cardiovascular disease (CVD) is currently one of the major contributors to the global burden of disease. Combination treatments to promote a maximal reduction of the ratio between total cholesterol and high-density lipoprotein are currently the most effective way of preventing CVD. In this review, we assess the role of plant sterols and statins in CVD prevention. Statins have been used by millions of patients at high to moderate risk of CVD, while plant sterols are potentially available to whole populations in food products. The benefits and risks of each compound, as well as the combination, are discussed. PMID- 15816509 TI - Conivaptan Yamanouchi. AB - Yamanouchi is developing conivaptan, a diuretic and active vasopressin V1a and V2 antagonist, which has an aquaretic effect, for the potential treatment of hyponatremia and heart failure. In January 2004, Yamanouchi submitted an NDA in the US for injectable conivaptan for the treatment of hyponatremia and, in December 2004, the FDA issued approval, although additional safety data were requested. PMID- 15816510 TI - Deferasirox Novartis. AB - Novartis is developing a chelating agent, deferasirox, as a once-daily oral therapy for the potential treatment of chronic iron overload in patients needing blood transfusions. By December 2004, data from a phase III trial for iron overload had been released, and filing is expected in the US and EU in the first half of 2005. PMID- 15816511 TI - Ranolazine CV Therapeutics. AB - CV Therapeutics, under license from Roche (formerly Syntex), is developing ranolazine (Ranexa), a metabolic modulator and a partial fatty acid oxidation inhibitor, for the potential treatment of angina and acute coronary syndromes. By October 2004, enrollment in the phase III MERLIN TIMI-36 study was ongoing and the company anticipated completion by the end of the first quarter of 2005. PMID- 15816512 TI - Androgen and androgen receptor antagonist responsive primary African-American benign prostate epithelial cell line. AB - The generation of suitable in vitro models is critical for understanding the process associated with the development and progression of prostate cancer in high-risk African-American men. However, the generation of long-term human prostate epithelial cell lines derived from primary human prostate epithelium have been unsuccessful due to the absence of in vitro immortalization. We have successfully established an immortal human prostate epithelial cell line from primary benign tissues of African-American prostate cancer patients by using telomerase. The actively proliferating secondary African-American prostate epithelial RC-165N cells, derived from benign prostate tissue of a radical prostatectomy specimen, were transduced through infection with a retrovirus vector expressing the human telomerase catalytic subunit (hTERT). A high level of telomerase activity was detected in RC-165N/hTERT cells but not in RC-165N cells. RC-165N/hTERT cells are currently growing well at passage 50 whereas RC-165N cells senesced within passage 3. RC-165N/hTERT cells exhibit epithelial morphology. These immortalized cells showed no cell growth in soft agar, and no tumor formation in SCID mice. The RC-165N/hTERT cells express androgen-regulated prostate-specific homobox gene. NKX 3.1 and epithelial cell specific cytokeratin 8, androgen receptor (AR), prostate stem cell antigen and p16, but not PSA. AR protein was detected by Western blot analysis. PMID- 15816513 TI - Preparation and preclinical assessment of folate-conjugated, radiolabelled antibodies. AB - BACKGROUND: The folate receptor (FR) is frequently over-expressed on human cancer cells and may be a suitable target for radiopharmaceuticals. Because of FR expression in the kidneys, the rapidly renal clearing folate is not well suited as a carrier for therapeutic radionuclides. As an alternative, folate immunoglobulin conjugates were studied as potential carriers for radionuclides. MATERIALS AND METHODS: Two types of conjugate were evaluated: (i) folate conjugated to osteosarcoma antigen directed murine monoclonal antibodies TP-1 and TP-3 or (ii) folate conjugated to non-specific polyclonal human IgG (HIg6). These constructs were labelled with 211At or 125I. RESULTS: The folate-HIg6 radionuclide conjugate showed high affinity to immobilized folate binding protein and also to folate receptor-expressing cells. Folate conjugates of TP-1 and TP-3 had a selective binding in vitro to antigen-expressing tumor cells and also to cells expressing FR only, thus the folate antibody constructs possessed dual affinity binding. Comparisons between folate-conjugated and non-folated antibody in Balb/C mice showed that the folate did not markedly change the properties of the radiolabelled antibody. CONCLUSION: It was demonstrated that folate conjugated antibodies carrying therapeutic radionuclides have relevant properties for the targeting of tumor cells expressing FR. PMID- 15816514 TI - Tyrosine kinase inhibitor SU6668 inhibits peritoneal dissemination of gastric cancer via suppression of tumor angiogenesis. AB - BACKGROUND: Peritoneal dissemination of cancer involves several steps, including tumor cell attachment, invasion and growth in the peritoneum. Tumor angiogenesis is a prerequisite for the growth of disseminated tumor. Vascular endothelial growth factor (VEGF) and its receptor are major regulators of angiogenesis. PURPOSE: We examined the cytotoxic effects of SU6668, an inhibitor of VEGF tyrosine kinase receptors, on in vitro gastric cancer cell lines and human umbilical vascular endothelial cells (HUVEC); we also examined the antitumor effects of SU6668 on human gastric cancer cells administered intraperitoneally into nude mice. MATERIALS AND METHODS: Direct cytotoxicity to gastric cancer cells (TMK-1, MKN-45 and MKN-74) and normal cells (HUVEC) was determined by the MTT assay and the bromodeoxyuridine (BrdU) incorporation assay, with and without VEGF-evoked growth stimulation in vitro. TMK-1 cells were transplanted intraperitoneally into nude mice, followed by twice daily oral administration of SU6668 (200 mg/kg/day) for two weeks starting on the first day after transplantation. Both the number and the wet weight of disseminated peritoneal tumor nodules were assessed. RESULTS: In the MTT assay, SU6668 demonstrated low grade cytotoxicity to the cell growth of three gastric cancer cells, with a 50% inhibitory concentration (IC50) of 22.6 microg/ml for TMK-1, 31.8 microg/ml for MKN-45 and 26.7 microg/ml for MKN-74; HUVEC was sensitive to SU6668 with an IC50 of 8.9 microg/ml. In the BrdU assay, VEGF stimulated DNA synthesis in HUVEC, while the incorporation of BrdU was not affected by VEGF in gastric cancer cell lines. SU6668 inhibited VEGF-induced DNA synthesis in HUVEC, while BrdU incorporation of gastric cancer cell lines was inhibited by SU6668 without correlation to VEGF stimulation. Peritoneal dissemination of cancer in nude mice was significantly suppressed by SU6668 compared with a control group at the p<0.05 level. CONCLUSION: The mechanism of the antitumor activity of SU6668 may not involve direct toxicity to cancer cells, but may rather be an inhibitory effect on tumor angiogenesis, resulting in the inhibition of tumor dissemination in the peritoneum. PMID- 15816515 TI - Surface expression and CEA binding of hnRNP M4 protein in HT29 colon cancer cells. AB - Carcinoembryonic antigen (CEA) has been shown to participate in the progression and metastatic growth of colorectal cancer. However, its biological function remains elusive. Recently, we found that CEA protects colon cancer cells from undergoing apoptosis, suggesting a complex role that includes signal transduction activity. Additionally, it was reported that CEA binds to Kupffer cells and macrophages to a membrane-anchored homolog of heterogeneous nuclear protein M4 (hnRNP M4), which subsequently was named CEA-receptor (CEAR). Cytoplasmatic and membranous expression of CEAR in CEA-positive colon cancer tissues prompted us to analyze the CEA-CEAR interaction in HT29 colon cancer cells. Both, CEA and CEAR were found on the cell surface of HT29 cells, as demonstrated by confocal microscopy. Imaging analysis suggested co-localization and, thus, interaction of both molecules. To confirm this observation, immunoprecipitation experiments and Western blot analysis were performed and indicated binding of CEA and CEAR. Immunoprecipitation of CEA resulted in a pull down of CEAR. The pull down of CEAR correlated with the amount of CEA as demonstrated by ribozyme targeting of CEA. Finally, external treatment of HT29 cells with soluble CEA induced tyrosine phosphorylation of CEAR, suggesting a CEA-dependent role of CEAR in signal transduction. Future experiments will elucidate whether the CEA-CEAR interaction is involved in CEA's antiapoptotic role and mediates the prometastatic properties of CEA in colon cancer cells. PMID- 15816516 TI - Radionuclide imaging of tumor xenografts in mice using a gelatinase-targeting peptide. AB - Tumors express MMP-2 and MMP-9 gelatinases, which are involved in the formation of tumor vasculature. This suggests that a tumor and its surrounding neovasculature can be visualized by a sensitive gelatinase recognition method. We have studied tumor radioimaging using a gelatinase inhibitory peptide CTTHWGFTLC (CTT), which in a mouse model targets the tumor site following an intravenous injection. We determined a solution NMR structure of CTT and its retro-inversion peptide, and prepared 125I and 99mTc-labelled CTT peptide derivatives. Radiolabelled CTT inhibited gelatinases in vitro, and homed to a tumor xenograft in mice. In normal mice, CTT was instead rapidly cleared from the circulation mainly through the kidney and, after 24 h, no significant radioactivity was accumulated in healthy tissues. Gamma camera imaging of a primary tumor in live mice was obtained with double-labelled liposomes, which were coated with 99mTc CTT and encapsulated with 125I albumin. CTT also targeted liposomes to the lungs of tumor-bearing mice, which may indicate the existence of non-visible lung micrometastases. Our studies suggest that selective gelatinase-targeting compounds could be useful in the early detection and imaging of primary tumors and metastases. PMID- 15816518 TI - A comparison of response to cisplatin, radiation and combined treatment for cells deficient in recombination repair pathways. AB - The responses of cells with mutated DNA repair pathways were compared for cisplatin, radiation and combination treatments. The knockout of the nonhomologous endjoining (NHEJ) pathway resulted in increased radiation sensitivity, but no change in cisplatin response in the mouse cells and increased radiosensitivity but decreased cisplatin sensitivity in chicken cells. The mutation of the homologous recombination repair (HR) pathway through XRCC3 in CHO cells resulted in increased radiation and cisplatin sensitivity and to a lesser extent for the Rad54 knockout in the DT40 chicken cells. The combination treatments of cisplatin and radiation showed that inhibition of the HR repair pathway resulted in super additive effects while the inhibition of the NHEJ pathway in DT40 had no effect. In mouse cells the knockout of the NHEJ pathway resulted in reduced super additivity compared to the parental cell lines. These data show that radiation, cisplatin and combination treatment damage is affected differently by the various DNA repair pathways, which could have a range of effects on combination treatments in tumour cells expressing different levels of DNA repair in the various repair pathways. PMID- 15816517 TI - Preclinical studies comparing different bispecific antibodies for redirecting T cell cytotoxicity to extracellular antigens on prostate carcinomas. AB - INTRODUCTION: Bispecific antibodies (BiAbs) are used to enhance targeting of T cells and other cytotoxic agents to tumors while minimizing non-specific tissue toxicities. This study compares the targeting efficacy of 3 BiAbs derived from chemically heteroconjugating a T cell-directed monoclonal antibody (mAb) to 9184, 9187 or 9189, which are mAbs directed at extracellular antigens expressed on human prostate carcinoma cell lines. MATERIALS AND METHODS: 9184 (anti-Her2/neu), 9187 (anti-gp55) and 9189 (anti-gp42) were each heteroconjugated to anti-CD3 to produce BiAbs capable of binding to ("arming") anti-CD3 activated T cells (ATC) and redirecting their cytotoxicity to prostate cancer cells expressing the respective antigen. ATC from cancer patients and/or normal subjects were armed with each BiAb and tested in co-cultures with PC-3, DU 145, and LNCaP cells for binding, cytotoxicity, and cytokine secretion. RESULTS: All 3 tumor-directed mAbs bound to each of the prostate cancer cell lines. ATC armed with 9184Bi statistically augmented cytotoxicity directed at PC-3 and increased IFN-gamma, TNF-alpha, and GM-CSF secretion as well as induced IFN-gamma EliSpots above that seen for 9187Bi, 9189Bi, ATC alone or ATC armed with an irrelevant BiAb. 9184Bi armed ATC mediated significant cytotoxicity against LNCaP and DU 145 cells as well. When we armed ATC from 6 cancer patients with 9184Bi, 9184Bi markedly enhanced cytotoxicity of ATC from 5 of the 6 patients. CONCLUSION: Arming ATC with BiAbs augments cytotoxicity directed at prostate cancer lines expressing the target antigens. Arming with 9184Bi was the most effective at redirecting cytotoxicity at PC-3 cells and inducing cytokine secretion. As an alternative to mAb therapy with anti-HER2, the HER2 antigen may provide a suitable target for redirecting anti-cancer immune cells, immunobiologicals, or other agents to HRPC. PMID- 15816519 TI - A rapid HPLC method for the measurement of ultra-low plasma methionine concentrations applicable to methionine depletion therapy. AB - A rapid, sensitive and specific high-performance liquid chromatographic (HPLC) method for the separation and quantification of L-methionine in plasma has been developed. After derivatization of plasma amino acids with o-phthalaldehyde (OPA), a 50 microl sample was loaded on a reversed-phase Supelcosil LC-18-DB column (particle size 5 microm, 25 cm x 4.6 mm, 120A pores). A customized gradient program using tetrahydrofuran/methanol/0.1 M sodium acetate, pH 7.0, v/v/v=5/95/900 and methanol was used with detection by fluorescence. The elution time was 15 minutes, a 3-fold improvement over existing methods. The linearity was 1-100 microM. The limit of detection was 0.5 micromol/L, a 10-fold improvement over existing methods. The inter-assay CVs were 2-5%, and the intra assays CVs were 4-8%. The sensitivity and rapidity of this HPLC method is particularly applicable to determine the efficacy of methionine depletion therapy of cancer. PMID- 15816520 TI - Role of p21WAF1 in green tea polyphenol-induced growth arrest and apoptosis of oral carcinoma cells. AB - The cyclin-dependent kinase inhibitor p21WAF1 participates in cell growth, differentiation, and apoptosis. p21WAF1 can be induced by green tea polyphenol EGCG in several cancer cell types, but its role in the oral cancer cell response to EGCG is not known. We found that EGCG upregulates p21WAF1 in an oral carcinoma cell line, OSC2, by cDNA microarray. The current study determined the impact of siRNA-suppressed p21WAF1 and its response to EGCG on cell growth, DNA synthesis and apoptosis by RT-PCR, Western blot, BrdU incorporation, MTT and caspase 3 activity assays. Suppression of p21WAF1 by siRNA resulted in an accelerated cell growth and DNA synthesis, and increased cell viability. However, caspase 3 activity was not significantly inhibited. The evidence showed that p21WAF1 is involved in EGCG-induced growth arrest of OSC2 cells, which may facilitate caspase 3-mediated apoptosis. Thus, expression of functional p21WAF1 may promote phytochemical-mediated growth arrest and apoptosis in oral carcinoma cells. PMID- 15816521 TI - Gene transfer of inhibitor kappaB in human lung cancer cell line NCI-H460 inhibits tumorigenesis and angiogenesis in vivo. AB - BACKGROUND: Nuclear factor kappaB (NFkappaB) is an inducible and ubiquitously expressed transcription factor which is involved in cell survival, differentiation and growth and, thus, has also been implicated in tumor formation and development. Research on the effect of NFkappaB in inhibiting cancer cell growth, however, remains controversial. MATERIALS AND METHODS: We investigated the effects of overexpressed IkappaBalpha on the proliferation of the human lung cancer cell line H460 in vitro and in vivo using IkappaBalpha-expressing adenovirus. RESULTS: The results suggested that the infection of AdIkappaBalpha blocked NFkappaB activity in H460 cells and significantly inhibited cell proliferation by inducing apoptosis. An in vivo study showed the tumor incidence to be significantly lower in mice implanted with H460 cells infected with AdIkappaBa. For established H460 tumor, the intratumoral injection of AdIkappaBalpha also inhibited the tumor growth due to both a blockade of the NFkappaB activity and an inhibition of the VEGF expression. CONCLUSION: Adenovirus-mediated IkappaBalpha gene transfer is a promising cancer treatment strategy. PMID- 15816522 TI - The relationship between bone metastasis from human breast cancer and integrin alpha(v)beta3 expression. AB - BACKGROUND: Osteoclast activation plays an essential role in the development of bone metastasis (BM). Integrin alpha(v)beta3 mediates the attachment of osteoclasts to bone matrix, and it is overexpressed in bone-residing breast cancer cells. We studied BM from breast cancer in relation to its Integrin alpha(v)beta3 expression. MATERIALS AND METHODS: Integrin alpha(v)beta3 expression in 4 human breast cancer cell lines (MDA-MB-231, MDA-MB-435, MKL-4 and T-47D) was determined by immunohistochemistry, flow cytometry and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Integrin alpha(v)beta3 was expressed in MDA-MB-231 and MDA-MB-435, but not in MKL-4 or T-47D, using both immunohistochemistry and flow cytometry. By RT-PCR, positive bands for both alpha(v) and beta3 subunits were identified in MDA-MB-231 and 435, while only the alpha(v) subunit mRNA was present in MKL-4 and T-47D. BM was common in vivo following inoculation with MDA-MB-231 or 435, but not with MKL-4. CONCLUSION: Integrin alpha(v)beta3 expression appears to play a key role in the development of BM from breast cancer. PMID- 15816523 TI - Tumor-draining lymph nodes of primary lung cancer patients: a potent source of tumor-specific killer cells and dendritic cells. AB - BACKGROUND: The tumor-draining lymph node tissue (TDLT) of lung cancer patients generated killer cells specific to autologous tumor cells when cultured with low dose IL-2. This production of killer cells lasted as long as 2 months after the initiation of the culture (productive phase). Even after this productive phase, TDLT supported the generation of the killer cells when these were co-cultured with peripheral blood lymphocytes (PBL) from the same patients. We tried to analyze the mechanisms of this production of killer cells from TDLT. MATERIALS AND METHODS: TDLT, tumor tissues as well as PBL were obtained from primary lung cancer patients and cultured in vitro. Cell growth, cell surface markers and specific cytotoxic activity of the lymphocytes were examined. RESULTS: The majority of the cells from TDLT or TDLT+ PBL co-culture (TDL-Pb) were CD3 positive T cells (89-99%) and a 51Cr-releasing assay showed that these cells had a stronger cytotoxic activity against autologous tumor cells than cells from PBL cultured with IL-2. Their activity against allogeneic MHC incompatible target cells was not, however, elevated. Cytotoxic activity against autologous tumor cells was blocked by anti-HLA class 1 (52.0%), class 11 (47.9%) and CD8 (46.8%) antibodies, but not by anti-CD56 antibody. The treatment of TDLT with anti-CD8, CD4, CD80 and CD83 all together completely abrogated the ability of TDLT to generate killer cells, with one of these antibodies it did so partially, while treatment with anti-CD56 antibody failed to do so at all. CONCLUSION: These results collectively suggest that TDLT contains tumor antigen-pulsed DCs as well as precursors of specific killer T cells and gives rise to the generation of killer cells when cultured in a low dose of IL-2. PMID- 15816524 TI - ET-18-OCH3 inhibits the phosphorylation and activation of p70 S6 kinase in MCF-7 cells. AB - Alkyllysophospholipids (ALPs) inhibit the proliferation of epithelial cancer cells, and may achieve this by perturbing a number of intracellular signaling pathways. p70 S6 kinase (p70S6K) is a key intracellular signaling molecule in the regulation of cell proliferation. We therefore investigated whether ALPs inhibit p70S6K activity and, if so, whether this may be relevant in the mechanism of inhibition of cell proliferation by ALPs. In this study, we demonstrated that the prototypic ALP, 1-O-octadecyl-2-O-methyl-rac-glycerophosphocholine (ET-18-OCH3), inhibits the activation of p70S6K in MCF-7 cells but does not inhibit the activity of p70S6K. Inhibition of activation is achieved by preventing the phosphorylation of Thr389, Thr421, Ser424 and Ser411 residues of p70S6K, which are required for full activation of the kinase. ET-18-OCH3 inhibited insulin stimulated activation of MCF-7 cells, which is sensitive to the phosphoinositide 3-kinase inhibitor LY294002, and to the m-Tor inhibitor rapamycin. Phorbol ester induced activation of p70S6K which was sensitive to the m-Tor inhibitor but not the phosphoinositide 3-kinase inhibitor, was also inhibited by ET-18-OCH3. Hence the diminished phosphorylation of p70S6K by ET-18-OCH3 is a result of the inhibition of both phosphoinositide 3-kinase-dependent and -independent activation of p70S6K. The differential effects of ENE-OCH3, a phosphonocholine analog of ET-18-OCH3, on MCF-7 cell proliferation correlated with its effects on p70S6K activation. The data suggest that the inhibition of p70S6K activation of by ET-18-OCH3 contributes to the antiproliferative effects of ALPs in MCF-7 cells. PMID- 15816525 TI - Blockage of cyclin cdk's, PKC and phosphoinositol 3-kinase pathways leads to augmentation of apoptosis in drug-resistant leukemia cells: evidence for interactive effects of flavopiridol, LY 294002, roscovitine,wortmannin and UCN 01. AB - A mouse leukemia L1210 cell line (Y8), selected for resistance to deoxyadenosine, has a markedly altered phenotypic expression that includes loss of sensitivity to dATP as an allosteric inhibitor of ribonucleotide reductase, increased expression of c-myc, c-fos and WAF1/p21, but decreased expression of p53. In addition, the Y8 cells have a Very strong apoptotic response to a variety of agents under conditions in which the parental wild-type cells do not apoptose. In these studies, we show that flavopiridol (a cdk inhibitor) causes the Y8 cells to undergo apoptosis via a caspase-3 activation process. The apoptotic response to flavopiridol is markedly enhanced by LY294002. Data also show that the apoptotic response of the Y8 cells to roscovitine (a cdk inhibitor) is enhanced by UCN-01 (a PKC inhibitor). These data show that simultaneous blockage of specific pathways leads to increased apoptosis in the Y8 cells with essentially no effects on the parental wild-type L1210 cells. PMID- 15816526 TI - The effect of cyclooxygenase-2 (COX-2) inhibition on human prostate cancer induced osteoblastic and osteolytic lesions in bone. AB - BACKGROUND: The mechanism of bone formation by osteoblastic prostate cancer metastases is not well defined. Using knockout mice, it has been demonstrated that prostaglandins produced by COX-2 are critical for fracture repair. Therefore, our aim was to determine if COX-2 plays a role in the bone formation in osteoblastic prostate cancer metastases in bone. MATERIALS AND METHODS: We assessed the influence of pharmacologic COX-2 inhibition in a SCID mouse intratibial injection model of bone metastasis using two human prostate cancer cell lines that produce either osteoblastic lesions (LAPC-9) or osteolytic lesions (PC-3, negative control). SC-58236, a COX-2 specific inhibitor, was used at a dose of 3 mg/Kg intraperitoneally 3 times per week in the Treatment groups for 8 weeks until sacrifice. RESULTS: Western blot for COX-2 demonstrated that LAPC-9 cells expressed high levels of COX-2 while PC-3 cells did not. Treatment with SC-58236 significantly reduced the size of osteoblastic lesions after LAPC-9 injection based on both radiographic and histomorphometric criteria compared to the control group. In contrast, large osteolytic lesions were seen in both control and SC-58236 treated animals after PC-3 cell injections. The results of this study indicate that COX-2 inhibition can decrease the size of osteoblastic lesions produced by LAPC-9, a human prostate cancer cell line that expresses high levels of COX-2. This treatment had no effect on the osteolytic activity of PC-3 cells. CONCLUSION: These findings suggest that the progression of osteoblastic metastases induced by human prostate cancer cells may be limited by COX-2 inhibitors. PMID- 15816527 TI - In vitro antimutagenicity of capsaicin toward heterocyclic amines in Salmonella typhimurium strain TA98. AB - Capsicum fruits are widely consumed as a component of the human diet. Capsaicin is the principle substance responsible for their hot, pungent taste. Heterocyclic amines (HCAs) are formed during cooking of meats and are mutagenic/carcinogenic compounds. In this study, we looked at whether capsaicin showed anti-mutagenic effects toward HCA-induced mutagenesis in Salmonella typhimurium TA98 when incubated with 0.5 mg liver S9 protein from rat, hamster and human. The HCAs used were Trp-P-2, Glu-P-1 and PhIP. Capsaicin, at non-toxic amounts of 0.25 and 0.5 micromole/plate, expressed a dose-dependent inhibition of the mutagenicity of Glu P-1 and PhIP when they are metabolically activated by rat, hamster and human liver S9 and of Trp-P-2 when activated by rat and hamster liver S9. In contrast, capsaicin enhanced the mutagenicity of Trp-P-2 in TA98 when incubated with human liver S9. The lack of consistency in the anti-mutagenic action of capsaicin toward HCAs is puzzling and currently unresolved. PMID- 15816528 TI - Pituitary tumor-transforming gene (PTTG): a novel target for anti-tumor therapy. AB - BACKGROUND: The proto-oncogene pituitary tumor-transforming gene (PTTG) is described as abundantly over-expressed in a variety of neoplasms with impact on neovascularization and tumor invasiveness. Based on these findings, we raised the question as to whether PTTG is a target for an anti-tumor therapy. MATERIALS AND METHODS: To investigate the impact of PTTG depletion on the human cervical cancer cell line HELA-S3, we used antisense-oligodesoxynucleotides (antisense-ODNs) and subjected them to in vitro transfection experiments. The PTTG mRNA level was determined by RT-PCR (45 cycles) and PTTG protein levels were determined by Western blot experiments. The impact of PTTG depletion on cell growth was determined 24, 48 and 72 hours post transfection by the trypan blue exclusion method. RESULTS: We found an antisense-ODN effective in down-regulation of PTTG mRNA and protein level. Furthermore, transfection resulted in a significant growth inhibitory effect as well as an increased level of apoptotic figures. CONCLUSION: Our studies revealed an anti-tumor potential for this target in cancer therapy. PMID- 15816530 TI - Roxithromycin inhibits angiogenesis of human hepatoma cells in vivo by suppressing VEGF production. AB - BACKGROUND: Recently, 14-member macrolide antibiotics such as Clarithromycin and Roxithromycin (RXM) have been shown to have anti-cancer and anti-angiogeneic effects. However, it is not fully understood whether and how RXM suppresses angiogenesis in human hepatoma, which is a well-known hypervascular tumor. MATERIALS AND METHODS: In the present study, we examined the effects of RXM on tumor angiogenesis in the human hepatoma cell line, HepG2. In vivo, angiogenesis was examined using a mouse dorsal air sac model. RESULTS: The inhibitory effect of RXM was dose-dependent and the angiogenesis index of 100mg/kg/day of RXM administered intraperitoneally twice a day was significantly lower than the control. Next, we examined the effect of RXM on vascular endothelial growth factor (VEGF) mRNA expression and its protein level in HepG2 cells. When 100 microM of RXM were added, VEGF mRNA expression in HepG cells was inhibited and its protein level reduced. CONCLUSION: These results suggest that RXM inhibits tumor angiogenesis in human hepatoma, and that VEGF alteration may be involved in the mechanism of this inhibitory effect. Because RXM is widely used in clinical practice, it may represent an effective new strategy for human hepatoma therapy. PMID- 15816529 TI - Wogonin inhibits N-acetyltransferase activity and gene expression in human leukemia HL-60 cells. AB - It is well documented that arylamine carcinogens are N-acetylated by cytosolic N acetyltransferase (NAT) enzyme. NAT plays an important role in the metabolizing of those arylamine compounds. 2-Aminofluorene (AF) is an arylamine carcinogen which has been demonstrated to induce carcinogenesis in laboratory animals. Our previous study has shown that a human promyelocytic leukemia cell line, HL-60, displays NAT activity. The purpose of the present study was to determine whether or not wogonin could affect the N-acetylation of AF in HL-60. N-acetylated and non-N-acetylated AF were determined by using high performance liquid chromatography. Wogonin displayed a dose-dependent inhibition of NAT activity in cytosols and intact cells. Wogonin also decreased AF-DNA adduct formation in these cells. The effects of wogonin on the NAT enzymes levels were also examined by Western blotting and flow cytometry and the changes of NAT gene expression were examined by polymerase chain reaction (PCR) and cDNA microarray. The results demonstrated that wogonin inhibited NAT1 mRNA gene expression and the level of NAT enzyme in HL-60 cells. This is the first demonstration that wogonin affects human leukemia cells' NAT activity in vitro. PMID- 15816531 TI - Apoptosis of human prostate androgen-dependent and -independent carcinoma cells induced by an isopropanolic extract of black cohosh involves degradation of cytokeratin (CK) 18. AB - BACKGROUND: The inhibitory effects of black cohosh extracts (Cimicifuga syn. Actaea racemosa L.) on the proliferation of human breast cancer cells were reported recently. In this study, we turned examined another hormone-dependent, epidemiologically important tumor disease, prostate cancer. The cell growth inhibitory effect of an isopropanolic extract of black cohosh (iCR) on androgen sensitive LNCaP and androgen-insensitive PC-3 and DU 145 prostate cancer cells was investigated. MATERIALS AND METHODS: The cytotoxic effect of the extract was determined by WST-1 assay. Apoptosis was determined by the appearance of apoptotic morphology, annexin V-FITC adherence and caspase activation. Cytokeratin (CK) 18 degradation was identified with M30 monoclonal antibody. RESULTS: Regardless of their hormone sensitivity, the growth of prostate cancer cells was significantly and dose-dependently down-regulated by iCR. The drug concentration producing 50% cell growth inhibition in all cell lines after 72h lay between 37.1 and 62.7 microg/ml. Increases in the level of M30 antigen of approximately 1.8-, 5.9- and 5.3-fold over untreated controls were observed in black cohosh-treated PC-3, DU 145 and LNCaP cells, respectively, with the induction of apoptosis being dose- and time-dependent. CONCLUSION: Black cohosh extract kills human hormone-responsive or-unresponsive prostate cancer cells by induction of apoptosis and activation of caspases. This finding suggests that the cell's hormone responsive status is not an important determinant of the response to the extract and that iCR extract may represent a novel therapeutic approach for the treatment of prostate cancer. PMID- 15816532 TI - Optimal drug and gene delivery in cancer cells by ultrasound-induced cavitation. AB - BACKGROUND: Lack of efficient drug and gene delivery is one of the major problems of cancer chemo- and bio-therapy. This work is focused on optimization of ultrasound-induced delivery of model macromolecular anti-cancer drugs and DNA in human cancer cells. MATERIALS AND METHODS: FITC dextrans simulated antisense oligonucleotides (10 kDa), antibodies (70 kDa), and genes (2000 kDa). Optimization of ultrasound frequency, intensity, duty cycle, time of irradiation, and concentration of Optison (ultrasound contrast agent) was performed. RESULTS: Optimal parameters provided in the MCF7 cell line 73.5+/-3.3%, 72.7+/-0.9%, and 62.7+/-2.1% delivery of 10-kDa, 70-kDa, and 2000-kDa macromolecules, respectively, 36.7+/-4.9% of cell transfection, while dead cell count was only 13.5+/-1.6%. Statistically significant drug delivery and transfection was obtained in all tested cell lines. CONCLUSION: These results suggest that optimized treatment parameters provide efficient drug and gene delivery in cancer cells and could be used for further in vivo and in vitro experiments. PMID- 15816533 TI - Paradoxical effect of cytosine arabinoside on mouse leukemia cell line L1210 cells. AB - We investigated the effects of 1-beta-D-arabinofuranosylcytosine (ara-C) on the growth of murine leukemic L1210 cells, which were cultured with high (2.0 x 10(3) ng/ml), middle (100 ng/ml) and low doses (5.0 ng/ml) of ara-C. In the analysis by flow cytometry, high dose ara-C arrested the cell cycle in the G0/G1-phase. Middle and low doses ara-C induced a block in the S-phase, that was not completely blocked by the low dose. Analysis of DNA fragmentation revealed that ara-C dose-dependently induced apoptosis, which was only slightly induced by the low dose. We measured activities of cellular thymidylate synthase (TS) and thymidine kinase (TK) after 24-h culture. Low and middle doses, but not high dose ara-C markedly enhanced TS activity to 2.9- in low and 5.3-fold in middle doses ara-C, and TK activity to 1.3- in low and 2.2-fold in middle doses, respectively, compared with those of the control. The cells accumulated in the S-phase by 48-h culture with low dose ara-C and markedly proliferated compared to that of the control in ara-C-free medium. These results indicate that non-high dose ara-C enhances DNA-synthesizing enzyme activities in L1210 cells, and withdrawal of the non-high dose ara-C results in paradoxical cell proliferation. Thus, daily intramuscular injections with an insufficient dose of ara-C may induce cells into S-phase, resulting in the proliferation of leukemic cells. PMID- 15816534 TI - Hydroquinone-induced apoptosis in HL-60 cells. AB - To clarify the mechanisms by which hydroquinone (HQ; 1,4-benzenediol) produces apoptosis, HQ-induced cytotoxicity, intemucleosomal DNA fragmentation, activation of superoxide dismutase (SOD), expression of Mn and Cu/ZnSOD mRNA and activation of caspase-3, -8 and -9 were investigated in the human promyelocytic leukemic cell line HL-60. Electrophoresis and activity staining of the SOD-enriched fraction showed that HQ reduced MnSOD activation more than Cu/ZnSOD activation, suggesting that it induces mitochondrial dysfunction at an early stage of apoptosis. Furthermore, the expression of MnSOD mRNA was suppressed to a greater extent than that of Cu/ZnSOD mRNA, implying that HQ causes apoptosis by inhibiting MnSOD induction. Release of cytochrome c and activation of procaspase 3 and -9, but not of procaspase-8, occurred more rapidly (as early as 6 h) in HQ treated cells, suggesting that HQ activates the intrinsic pathway of apoptosis. Addition of the antioxidant N-acetyl-L-cysteine (NAC) significantly reduced the cytotoxicity of HQ. At a concentration that was cytotoxic to 50% of the cells (approximately 0.05 mM), HQ activated caspase-3; this effect was reduced in the presence of NAC. Interestingly, higher concentrations of HQ (0.1-0.2 mM) caused direct cell death; however, when combined with 5 mM NAC, the activation of caspase-3 was strongly enhanced, suggesting the promotion of apoptosis. The activation of caspase-3 by HQ/NAC combinations suggests that NAC, a precursor of intracellular glutathione synthesis, acts as a co-catalyst during HQ-induced apoptosis. PMID- 15816535 TI - Involvement of angiopoietins in cancer progression in association with cancer cell--fibroblast interaction. AB - PURPOSE: The role of angiopoietins (ANGs) in cancer progression is a new field of research. We examined the patterns of expression of ANGs in human renal cancer tissues (n =12) and investigated their roles in cancer progression in vitro and in vivo. METHODS AND RESULTS: In normal renal tissue, the expression of ANG-1 was apparent in the glomerular capillaries and podocytes as well as in the endothelial cells of vessels, whereas we observed no expression of ANG-2. In contrast, in tumor tissue, dense, diffuse expression of ANG-1 was apparent in the fibroblasts, but not in the cancer cells. Intense ANG-2 expression was observed in the cancer cells themselves as well as in the endothelial cells, where its expression was restricted to small vessels or neoplastic capillaries and not mature vessels. Secondly, we investigated the influence of fibroblasts on the production of angiopoietins in cancer cells, using the human renal cancer cell lines SN12 and SN-PM6 and a human dermal fibroblast cell line (HDF). We examined the in vitro production of angiopoietins in these cell lines, in either monoculture of each cell line or co-culture of cancer cells and fibroblasts. Immunohistochemical study demonstrated marked production of ANG-1 in fibroblasts and ANG-2 expression in cancer cells when we performed co-culture, but no expression of either in each monoculture. Western blot analysis confirmed these results, showing marked expression of ANGs in co-cultured cells, but not in each monoculture. CONCLUSION: Fibroblasts may influence cancer progression by promoting neoplastic angiogenesis, and ANGs are profoundly involved in this process through their association with the carcinoma cell-fibroblast interaction. PMID- 15816536 TI - HBME-1 expression in follicular tumor of the thyroid: an investigation of whether it can be used as a marker to diagnose follicular carcinoma. AB - BACKGROUND: HBME-1 has been recognized as a useful marker for diagnosing thyroid carcinoma. In this study, we investigated whether it has a diagnostic value for discriminating follicular carcinoma from adenoma. MATERIALS AND METHODS: We investigated HBME-1 expression in 138 follicular carcinomas, 155 follicular adenomas, 98 adenomatous nodules and 37 papillary carcinomas, using anti-HBME-1 monoclonal antibody. RESULTS: HBME-1 was positive in 60.9% of follicular carcinoma and the incidence was significantly higher (p<0.0001) than that of follicular adenoma, 30.3%. In adenomatous nodules, only 17.3% were classified as positive, which was lower even than that of follicular adenoma (p=0.0257). All papillary carcinomas examined were positive for HBME-1. We calculated the positive predictive value of HBME-1 in discriminating follicular carcinoma from adenoma as 64.2%. CONCLUSION: These results suggest that, although HBME-1 contributes to the diagnosis of papillary carcinoma, it could not be applied in the preoperative diagnosis of follicular carcinoma, for example, using fine needle aspiration biopsy samples. PMID- 15816537 TI - Identification of metastasis-associated genes in prostate cancer by genetic profiling of human prostate cancer cell lines. AB - OBJECTIVES: Prostate cancer (PCa) is a heterogeneous tumour entity with known interindividual differences in biological behaviour regarding tumour aggressiveness and metastatic potentiaL To date, the prediction of the metastatic status of patients with PCa has not been possible. To identify the molecular causes behind these differences, the gene expression profiles of two cell lines (LNCaP and LNCaP C4-2) with different metastatic potentials were examined using DNA microarray technology. MATERIALS AND METHODS: LNCaP and LNCaP C4-2 cells were cultured under standard conditions. RNA was isolated using Trizol extraction. After processing the total RNA according to the manufacturer's instructions, we performed Affymetrix GeneChip analysis with HG-U133A chips. Data analysis was performed using NetAffx, dChip, GenMAPP and OMIM software. RESULTS: After statistical evaluation of the raw data, we obtained a set of 158 differently expressed probe sets in the LNCaP and LNCaP C4-2 cells. The search for genes associated with proliferation, cell metabolism, growth factors, metastatic potential and tumour progression in this list revealed a number of 42 differently expressed probe sets. The comparison of this list of probe sets with the literature resulted in a list of 14 differently expressed genes which could well contribute to the metastatic potential and progression of PCa. Of these 14 genes only 6 (Cip1, IGF-1, NK4, CXCL 12, ILGF2R, RHOE) have already been associated with PCa, whereas the other 8 genes (FSTL-1, SOCS-2, Midkine, Thrombospondin 1, Secretory leukocyte protease inhibitor, Desmoglein 2, MLT 1, PTPRF) had not been previously related to PCa. CONCLUSION: DNA microarray technology offers the possibility of screening a large number of genes with regard to alterations in the expression level or mutations. In this study, we identified 14 genes that are most probably associated with the higher metastatic potential of LNCaP C4-2 cells as compared to LNCaP cells. Eight of these 14 genes are potential new molecular markers for assessing the metastatic potential of PCa, or may serve as therapeutical targets. PMID- 15816538 TI - Serum enkephalin-degrading aminopeptidase activity in N-methyl nitrosourea induced rat breast cancer. AB - BACKGROUND: Breast cancer is the most frequent spontaneous malignancy diagnosed in women in the western world, although no specific etiological agent(s) or the mechanism responsible for the initiation of the disease has been identified as yet. Enkephalins (Leu5-enkephalin and Met5-enkephalin) (ENK) act in the breast in different ways such as modulating esteroid receptors and proteases secretion. ENK are hydrolyzed by specific enzymes, leading to their inactivation, such as the enkephalin-degrading tyrosyl aminopeptidase (EDA). Breast tumours induced in rats by administration of N-methyl-nitrosourea (NMU) constitute a useful tool for dissecting the multistep process of carcinogenesis, which involves initiation, promotion and progression. The aim of the present work was to analyse EDA activity (E.C: 3.4.11.-) in serum of rats with mammary tumours induced by NMU, to evaluate the potential value of this activity as a biological marker of the carcinogenesis process, and the putative role of ENK in the promotion and progression of the disease. MATERIALS AND METHODS: Tumours were induced by intraperitoneal injection of three doses of NMU at 50, 80 and 110 days after birth. Serum EDA was measured fluorimetrically using tyrosyl-beta-naphthylamide as substrate. RESULTS: The increase found in EDA activity suggests the existence of decreased serum circulating levels of ENK in rat with mammary tumours induced by NMU. CONCLUSION: Although the exact role of ENK in breast cancer initiation, promotion and/or progression remains unknown, our results suggest that changes in EDA activity might play an important role in the origin and evolution of breast cancer. PMID- 15816539 TI - Antiangiogenesis and anticancer efficacy of TA138, a novel alphavbeta3 antagonist. AB - BACKGROUND: Angiogenesis is a complex process involving endothelial cell migration, proliferation, invasion, and tube formation. Inhibition of these processes might have implications in various angiogenesis-mediated disorders. MATERIALS AND METHODS: The antiangiogenic efficacy of the novel alphavbeta3 antagonist TA138 was examined using in vivo and in vitro model systems. RESULTS: The in vitro studies demonstrated the ability of TA138 and RP747 (conjugated TA138) to inhibit endothelial cell migration toward vitronectin, with an IC50=0.04 and 0.045 microM, respectively. Furthermore, utilizing the chick chorioallantoic membrane models, TA138 inhibited basic fibroblast growth factor induced neovascularization. CONCLUSION: TA138 might be a useful tool for the inhibition of angiogenesis associated with human tumor growth, or other pathological neovascularization processes. RP747 demonstrated antitumor efficacy in 1 spontaneous tumor model (c-neu oncomouse model, alphavbeta3 positive cells) and in 1 xenograft model (HCT116 human tumor colon carcinoma, alphavbeta3 negative cells) injected subcutaneously into nude mice. PMID- 15816540 TI - Digoxin inhibits neuroblastoma tumor growth in mice. AB - BACKGROUND: Epidemiological data suggest a more favorable outcome of breast carcinoma in women taking cardiac glycosides. This study investigated whether digoxin could inhibit tumor growth in mice. MATERIALS AND METHODS: Tumor growth experiments were done in mice grafted with the neuroblastoma cell lines SH-SY5Y, Neuro-2a, colonic cancer cells (LS174T) or Lewis lung cancer cells (LLC). Angiogenesis inhibition was investigated in vitro on fibroblast growth factor-2 (FGF-2)-stimulated bovine endothelial cell (BCE) growth and in vivo in the chick chorioallantoic membrane (CAM) assay. RESULTS: SH-SY5Y and Neuro-2a grafts were inhibited by 44% (p=0.008) and 19% (p=0.007), respectively, whereas the colonic cancer xenografts and LLC syngrafts were less responsive. The neuroblastoma specificity was confirmed in vitro. Digoxin also inhibited angiogenesis in the CAM assay and the BCE cell survival in vitro was 50% at 53 ng/ml. CONCLUSION: Our data suggest that digoxin may be a specific neuroblastoma growth inhibitor and an unspecific inhibitor of angiogenesis. PMID- 15816541 TI - Comparative assessment of the functional p53 status in glioma cells. AB - BACKGROUND: p53 is the most frequently mutated gene in human cancers and its functional integrity is an important predictor of treatment response and clinical outcome. The majority of mutations found in different types of cancer cluster within the DNA binding domain encoded by exons 5-8. In clinical specimens the functional status of p53 is, therefore, often evaluated by direct mutation analysis of exons 5-8 or indirectly by immunostaining and evaluation of the subcellular localization pattern or protein accumulation. MATERIALS AND METHODS: In a panel of glioma cell lines, the status of the P53 gene was analyzed by temperature gradient gel electrophoresis (TGGE) of exons 5-8 and direct sequencing of all p53 exons. The nuclear accumulation of p53 in unstressed cells was assessed by immunostaining. These data were correlated with stress induction of the p53 protein, nuclear translocation and a direct determination of the transcriptional activity of endogenous p53 protein and induction of p53 target genes. RESULTS: Our analysis demonstrated that a p53 gene mutation analysis limited to exons 5-8 and analysis of immunostaining patterns can not serve as reliable predictors of functional p53 in tumor cells. Conversely, in some presumably rare cases, the transcriptional activity of p53 may be retained in tumor cells in the presence of a mutation and a pathological immunostaining pattern. In our analysis, the constitutive dephosphorylation at Ser 376 correlated with the nuclear accumulation of p53, but not with the transcriptional activity of the protein. This suggests that constitutive dephosphorylation at Ser376 may be one of the factors determining stabilization of mutant and wild type p53, which is frequently observed in glial tumors. CONCLUSION: The incidence of a dysfunctional p53 protein in gliomas may be higher than expected, based on a single parameter evaluation by mutation analysis of exons 5-8 or assessment of p53 accumulation and subcellular localization by immunostaining. PMID- 15816542 TI - Growth inhibitory effect of celecoxib and rofecoxib on human colorectal carcinoma cell lines. AB - BACKGROUND: Epidemiological studies have revealed a protective effect of NSAIDs, which principally target cyclooxygenase (COX)-1 and COX-2, on the development of colorectal cancer. Increased expression of COX-2 was shown in colorectal adenocarcinoma. However, some effects were shown to be COX-independent. Here, we compared two selective COX-2 inhibitors for their effect on the growth of colorectal tumour cells in vitro. MATERIALS AND METHODS: Fifteen tumour cell lines were characterized for COX-1 and COX-2 expression by Western blot and RT PCR. The effect of celecoxib and rofecoxib on their growth was assessed by staining of DNA with crystal violet. RESULTS: COX-2 expression varied among cell lines, whereas COX-1 was always expressed. Rofecoxib displayed a limited dose related effect on cell proliferation, whereas celecoxib strongly inhibited cell growth at high concentrations. Both effects appeared COX-2-independent. CONCLUSION: Rofecoxib, which is devoid of apoptotic effect at high concentration but efficient at pharmacological concentrations, revealed a potential new mechanism of action of NSAIDs towards colorectal cancer. PMID- 15816543 TI - 1,24(S)-dihydroxyvitamin D2, an endogenous vitamin D2 metabolite, inhibits growth of breast cancer cells and tumors. AB - BACKGROUND: 1,24-Dihydroxyvitamin D2 (1,24(OH)2D2) is a naturally occurring metabolite of vitamin D2 with low calcemic activity and potent antiproliferative activity. We evaluated the activity of 1,24(OH)2D2 in breast cancer models. MATERIALS AND METHODS: The antiproliferative activity of 1,24(OH)2D2 was quantitated against human and murine breast cancer cell lines. The antitumor activity of 1,24(OH)2D2 was quantitated using MCF-7 xenografts in nude mice. RESULTS: 1,24(OH)2D2 inhibited growth of vitamin D receptor (VDR)-positive, but not VDR-negative, breast cancer cells. 1,24(OH)2D2 (10 microg/kg or 50 microg/kg) reduced MCF-7 xenograft growth by 50% after five weeks. Tumor morphology in treated animals was consistent with replacement of epithelial cells by stromal tissue. Mice treated with 1,24(OH)2D2 showed no loss of body weight, hypercalcemia or kidney calcification. CONCLUSION: 1,24(OH)2D2 inhibits growth of breast cancer cells via VDR-dependent mechanisms; its complete lack of toxicity and significant antitumor activity supports further development for chemotherapeutic applications. PMID- 15816544 TI - Progesterone induces BRCA1 mRNA decrease, cell cycle alterations and apoptosis in the MCF7 breast cancer cell line. AB - BACKGROUND: Inherited mutations of the BRCA1 gene are responsible for hereditary breast and ovarian cancer syndrome. However, little is known of how disruption of BRCA1 functions preferentially increases cancer risk in hormone-dependent organs. We aimed to study whether BRCA1 was regulated by progesterone in the MCF7 breast cancer cell line. MATERIALS AND METHODS: MCF7 breast cancer cells were incubated with 10(-4) or 10(-10) M progesterone for 24 or 48 hours. BRCA1 expression, proliferation and apoptosis were analysed. RESULTS: 10(-4) M progesterone decreased cell proliferation, cell cycle progression and induced apoptosis. In addition, BRCA1 and cyclin A mRNA decreased. In contrast, none of these effects were observed in MCF7 cells incubated with 10(-10) M progesterone. CONCLUSION: The down-regulation of BRCA1 in MCF7 cells incubated with 10(-4) M progesterone seems to be a consequence of cell cycle alterations rather than a direct effect of the hormone on BRCA1. PMID- 15816545 TI - Action of fenretinide (4-HPR) on ovarian cancer and endothelial cells. AB - BACKGROUND: Fenretinide (4-HPR) is a synthetic retinoid that has been reported to inhibit the growth of cancer cell lines in vitro. MATERIALS AND METHODS: We examined the effect of 4-HPR on ovarian cancer (OVCAR-5) cell proliferation, viability and invasion using standard techniques. We also examined the action of 4-HPR on the actin cytoskeleton using immunocytochemistry, and on phosphorylation of focal adhesion kinase (FAK) using immunoprecipitation and phosphotyrosine immunoblotting. We then examined the activity of 4-HPR on endothelial cells using the tube formation assay on Matrigel. RESULTS: 4-HPR inhibited OVCAR-5 cell proliferation and viability at concentrations higher than 1 microM, with 70-90% growth inhibition at 10 microM. 4-HPR (1 microM) significantly inhibited OVCAR-5 invasion after 3 days preincubation. In view of the importance of the cytoskeleton in cell motility, we examined the action of 4-HPR on the actin cytoskeleton and on FAK phosphorylation. In OVCAR-5 cells treated with 1 mM fenretinide for 3 days, actin cytoskeleton stress fibers were disrupted and FAK tyrosine phosphorylation was elevated dose-dependently. Endothelial cells treated with 1 microM 4-HPR failed to form tubes, but formed small cellular aggregates. CONCLUSION: Fenretinide has anti-tumor activity by acting on the actin cytoskeleton and by regulating FAK tyrosine phosphorylation. 4-HPR also inhibits endothelial cell tube formation, a major step in angiogenesis. PMID- 15816546 TI - Consequences of chemoresistance for the herpes simplex virus thymidine kinase/ganciclovir-induced bystander effect in a human small cell lung cancer cell line model. AB - This paper focuses on the influence of chemoresistance on the herpes simplex virus (HSV-tk)/ganciclovir (GCV)-induced bystander effect (BE), as studied in a human small cell lung cancer (SCLC) cell line (GLC4) and its sublines with in vitro acquired resistance to adriamycin (GLC4/ADR), mitoxantrone (GLC4/MITO) and cisplatin (GLC4/CDDP). Chemoresistance for adriamycin, mitoxantrone and cisplatin significantly changed GCV sensitivity. A significant BE was found in all GLC4 cell lines. Compared to the parental GLC4 cell line, the BE was significantly higher only for the GLC4/ADR cell line. No expression of the nucleoside transporters MRP4 and MRP5 was detected. In all cell lines expression of connexin 43 was found, but modulation of gap junctional intercellular communication (GJIC) by 18-alpha-glycyrrhetinic acid did not significantly change the BE in any of the GLC4 cell lines. In conclusion, chemoresistance can influence the HSV-tk/GCV induced BE, which seems not to be related to differences in MRP4/MRP5 expression or to differences in GJIC. PMID- 15816547 TI - Protein expression and gene copy number analysis of topoisomerase 2alpha, HER2 and P53 in minimally invasive urothelial carcinoma of the urinary bladder--a multitissue array study with prognostic implications. AB - BACKGROUND: Due to their variable clinical course, there is a need for new prognostic parameters in minimally invasive (stage T1) bladder carcinomas. MATERIALS AND METHODS: Using multitissue arrays, protein expression and gene copy numbers of topoisomerase (TOP2alpha), HER2 and p53 were investigated by immunohistochemistry and by fluorescence in situ hybridization (FISH) in 73 T1 tumors. The results were compared with tumor recurrence and progression. RESULTS: The median TOP2alpha and p53 index was 21% (range, 3-59%) and 7% (range, 0-93%), respectively. HER2 overexpression (score 3+) was detected in 9 cases (12%). High TOP2alpha and p53 indices and HER2 overexpression were significantly associated with earlier tumor recurrence, but not with earlier tumor progression. While TOP2alpha and p53 gene amplification was detected in no case, 5 cases (8%) showed HER2 gene amplification, which was related to HER2 3+ score in 4 cases. Loss of TOP2alpha, HER2 and p53 gene was observed in 4 (8%), 8 (13%) and 6 cases (12%), respectively. By univariate analysis, TOP2alpha index (p=0.0267), HER2 score (p =0.028) and p53 index (p=0.0188) were significantly and loss of TOP2alpha gene (p=0.0575) tendentially correlated with tumor recurrence, while loss of HER2 gene (p=0.069) and loss of p53 gene (p=0.0587) were tendentially correlated with tumor progression. In a multivariate analysis, which also included tumor grade and T1 substage, TOP2alpha index (p=0.043) and p53 index (p=0.02) were identified as independent predictors of tumor recurrence and loss of p53 gene (p=0.012) and T1 substage (p=0.029) as independent predictors of tumor progression. CONCLUSION: Immuno-histochemical TOP2alpha and p53 staining as well as FISH analysis of p53 gene copy numbers and T1 substaging are helpful means of providing additional information on the biological behavior of T1 transitional cell carcinomas. PMID- 15816548 TI - Cyclin D1 overexpression is associated with estrogen receptor expression in Caucasian but not African-American breast cancer. AB - BACKGROUND: African-American (AA) breast cancer patients consistently show a shortened survival when compared with Caucasian patients. This worse prognosis is most likely due to a combination of socioeconomic factors and differences in tumor biology. Previous studies have demonstrated that cyclin D1 overexpression is strongly associated with estrogen receptor (ER) expression in breast cancer, but these series either included primarily Caucasian patients or did not specify race. MATERIALS AND METHODS. We analyzed 200 breast cancer cases obtained from AA and Caucasian patients who were matched on age, stage, ER status, and year of diagnosis. We examined expression levels of cyclin D1, p53, p27Kip1 (p27), and p21Cip1 (p21), and correlated their expression with ER status. RESULTS: Cyclin D1, p53, p27, and p21 expression rates were similar in matched cases of AA and Caucasian breast cancer (p values>0.05). However, cyclin D1 overexpression was significantly associated with ER status in only the Caucasian (p=0.0005), and not the AA cases (p=0.07). CONCLUSION: This finding suggests a novel biological difference, which may contribute to the more aggressive phenotype of AA breast cancer. PMID- 15816549 TI - Protection by antioxidants of copper-induced decline of proliferation and SOD activity. AB - The effect of Cu plate on the cellular function was investigated by two different methods: an extraction method (Method I) and a direct contact method (Method II). In Method I, the supernatant of the culture medium, which had been pre-incubated with Cu plate, was added to mouse macrophage-like Raw 264.7 cells. This supernatant dose-dependently inhibited the proliferation and nitric oxide (NO) production by lipopolysaccharide-stimulated Raw 264.7 cells. In Method II, human promyelocytic leukemic HL-60 cells in suspension were incubated with culture medium which contained Cu plate. The direct contact with Cu plate rapidly suppressed the proliferation and MnSOD and Cu/ZnSOD activities. The suppressed proliferation and SOD activity reverted to or exceeded the control level by sodium ascorbate, whereas N-acetyl-L-cysteine (NAC) only reactivated the proliferation, but not the SOD activity. ESR spectroscopy showed that contact with Cu plate slightly diminished the hydroxyl radical (generated by Fenton reaction), without affecting the intensity of NO (produced from NOC-7) and DPPH radical. The present study suggests that two representative antioxidants, such as sodium ascorbate and NAC, protect the cells from Cu-induced cytotoxicity via different mechanisms. PMID- 15816550 TI - Photochemical enhancement of gene delivery to glioblastoma cells is dependent on the vector applied. AB - BACKGROUND: Photodynamic therapy (PDT) and gene therapy protocols are separately under clinical evaluation for treatment of brain malignancies. Here, the potential of a novel combination technique, photo-induced delivery of macromolecules and genes to glioblastoma cells, is evaluated. MATERIALS AND METHODS: The photochemical effect on survival of GaMg and U-87Mg cells after incubation with the protein toxin gelonin, on transfection with a plasmid complexed to poly-L-lysine (PLL), and on transduction with adenovirus serotype 5 (Ad5) and adeno-associated virus type 5 (AAV5) vectors, were studied. RESULTS: Cytotoxicity of gelonin and gene transfer from plasmid/PLL complexes were considerably improved by photochemical treatment in both cell lines, while the light-inducible effect on Ad5 transduction was most pronounced in U-87Mg. For the first time, photochemical enhancement of AAV transduction is shown. A 4-fold increase in percentage positive cells was detected after photochemical treatment of AAV5-infected GaMg cells. However, in contrast to Ad5, AAV5 transduction of U 87Mg remained unaffected by light treatment, independently of viral dose, light dose and timing of the light treatment relative to the transduction period. CONCLUSION: Photochemical treatment is a versatile tool for macromolecular delivery to glioblastoma cells, however, the photochemical effect on gene transfer by viral vectors is highly dependent on the cell line and vector applied. PMID- 15816551 TI - Vaccination of apoptotic Fas ligand-expressing tumors decreased antitumor responses by enhanced production of immunosuppressive cytokines. AB - Expression of Fas ligand (FasL) in tumors produced antitumor effects by generating both inflammation and T cell-mediated immunity, although the Fas/FasL interaction induces an apoptotic process of Fas-positive activated T cells. Our previous study, however, showed that immunization of mice with ultraviolet (UV) treated FasL-expressing tumors rather induced immune suppression to the tumors, whereas mice rejected UV-untreated FasL-expressing tumors and developed protective immunity subsequently. Since dendritic cells (DCs) control tumor specific immune responses in vivo, we examined a possible role of DCs in the immune suppression induced. Administration of DCs that were co-cultured with UV treated FasL-expressing tumors did not influence the growth of parent tumors that were subsequently inoculated. Migration of immunocompetent cells into UV-treated FasL-expressing tumors was not significantly different from that into UV untreated FasL-expressing tumors. However, production of immunosuppressive but not T helper type 1 cytokines was enhanced when UV-treated FasL-expressing tumors were administered. These data collectively suggest that the immune suppression induced by UV-treated FasL-expressing tumors was not attributable to tolerance of DCs, but due to cytokine-induced suppression of cell-mediated immunity. PMID- 15816552 TI - Cytotoxic activity of azulenequinones against human oral tumor cell lines. AB - We investigated twenty-seven azulenequinone derivatives for their relative cytotoxicity against three human normal cell lines (HGF, HPC, HPLF) and four human tumor cell lines (HSG, HSC-2, HSC-3, HL-60). Parent 1,5-azulenequinone showed potent and some tumor-specific cytotoxicity. Halogenated derivatives of 1,5- and 1,7-azulenequinone showed potent cytotoxicity, but lower tumor-specific cytotoxicity. In contrast to other azulenequinones, amino derivatives such as 3 amino-1,5- and 1, 7-azulenequinones showed relatively lower cytotoxic activity. The 3-Phenoxy-1,5-azuleneqinone derivative showed higher cytotoxicity than the 3 phenoxy-1, 7-azulenequinone derivative. 1,5- and 1,7-Azulenequinones generally showed higher cytotoxicity, as compared with tropolones and azulene derivatives. 3- (3-Guaiazulenyl)-1, 5-azulenequinone [12] and 7-isopropyl-3- (4-methylanilino) 2-methyl- 1, 5-azulenequinone [24] showed a relatively higher TS value and induced apoptosis (internucleosomal DNA fragmentation, activation of caspases 3, 8 and 9) in HL-60 and HSC-2 cells, possibly via the activation of both mitochondria-independent (extrinsic) and -dependent (intrinsic) pathways. Western blot analysis showed that [24] slightly increased the intracellular concentration of pro-apoptotic proteins (Bad, Bax) in HSC-2 cells, whereas [12] was much less active. None of the twenty-seven azulenequinones showed anti-HIV activity. These results suggest [12] and [24] as possible candidates for future cancer chemotherapy. PMID- 15816553 TI - A sesquiterpenelactone from Inula britannica induces anti-tumor effects dependent on Bcl-2 phosphorylation. AB - BACKGROUND: The over-expression of the anti-apoptotic protein Bcl-2 in cancer is associated with resistance to chemotherapeutic drugs. The phosphorylation of Bcl 2 is one mechanism by which anti-microtubule agents, such as paclitaxel or docetaxel, may inactivate Bcl-2. Although initially active in clinical studies, current anti-microtubule agents are only temporarily effective and the discovery of new agents is warranted. MATERIALS AND METHODS: We isolated and identified two known sesquiterpenelactones, O, O-diacetylbritannilactone (OODABL) and O acetylbritaanilactone (OABL) from the flowers of the medicinal plant Inula britannica and studied their mechanism of anti-tumor effects. To determine the biological significance of Bcl-2 phosphorylation, we used a baby rat kidney (BRK p53) cell line that was transformed with EIA and a temperature-sensitive mutant p53. The BRK-p53 cell line was transfected with either a vector with wild type Bcl-2 or a vector in which Bcl-2 had mutations in the paclitaxel phosphorylation sites (pcDNA3.1 V5/His Bcl-2 S70, 87A). RESULTS: OODABL and OABL induced phosphorylation of Bcl-2 in breast, ovary and prostate cancer cell lines and induced G2/M cell cycle arrest. Using the BRK cells with mutant Bcl-2 (BRK-Bcl-2 mt) and control (BRK-Bcl-2-wt), we found that OODABL induced phosphorylation of Bcl-2 at sites similar to paclitaxel. Phosphorylation of Bcl-2 was important for OODABL-induced cytotoxicity, since the abrogation of phosphorylation in BRK-Bcl-2 mt cells decreased OODABL-induced cytotoxicity. CONCLUSION: We concluded that OODABL is cytotoxic in multiple tumor cell lines, and the cytotoxicity is dependent, at least in part, on the phosphorylation of Bcl-2. PMID- 15816554 TI - A novel chromatographic method for Ep-CAM mRNA detection in peripheral blood and bone marrow of patients with metastatic colorectal cancer. AB - BACKGROUND: The aim of our study was to detect Ep-CAM mRNA in peripheral blood (PB) and bone marrow (BM) samples of patients with metastatic colorectal cancer, using a novel chromatographic method (molecular strip) for the detection of PCR amplified product and to confirm the results by the standard method of agarose gel electrophoresis. PATIENTS AND METHODS: Thirty patients participated in the present study providing 27 PB and 26 BM samples. PB samples were also obtained from 20 healthy volunteers. PCR products were detected using the molecular strip and the results were confirmed by 1.5% agarose gel electrophoresis. RESULTS: In total 26 out of 27 blood samples (96%) and 19 out of 26 bone marrow samples (73%) were found positive for Ep-CAM expression using both methods, whereas none of the 20 healthy controls was found positive for the above marker. CONCLUSION: Ep-CAM mRNA is highly expressive in peripheral blood of patients with metastatic colorectal cancer. Its prognostic significance should be evaluated in patients with resectable carcinomas. The new strip detection method is highly specific for the PCR product under investigation, reliable, easy to handle, fast and safe for the user. PMID- 15816555 TI - Growth inhibitory effect of the somatostatin structural derivative (TT-232) on leukemia models. AB - TT-232 is a structural derivative of the natural signal inhibitory peptide somatostatin, with selective antiproliferative and anti-inflammatory properties. TT-232 activates SSTR receptors (primarily the SSTR-1), which leads to irreversible cell cycle arrest, followed by secondary induction of apoptosis. TT 232 has passed phase I clinical trials without toxicity and significant side effects. We examined the antiproliferative effect in vitro and the antitumor effect in vivo of TT-232 on leukemia cell lines. During in vivo experiments, we evaluated the therapeutic efficacy of TT-232 in various long-term administration routes; traditional injection versus infusion treatment via an inserted Alzet minipump on P-388 mice and HL-60 human leukemia models. Treatment with TT-232 started after development of the disease. In vitro, TT-232 inhibited the proliferation of P-388 mice lymphoid cells and HL-60 human promyelocytic leukemia cells in the range of 46%-97% with 24-hour treatment and 82%-100% with 48-hour treatment. Cells were treated with 30 microg/ml and 60 microg/ml dose of TT-232. With the same in vivo models, the best results were achieved when TT-232 was applied by infusion treatments. The infusion treatment with TT-232 produced 50% 80% inhibition of growth and resulted in 20%-40% long-term and leukemia-free survivors. TT-232 showed dose-, time- and administration mode-dependent antileukemia activity in vitro and in vivo, both on rodent and human models. Our results suggest that TT-232 is a promising new antileukemia agent. PMID- 15816556 TI - Preclinical safety, pharmacokinetics and antitumor efficacy profile of liposome entrapped SN-38 formulation. AB - BACKGROUND: SN-38, 7-ethyl-10-hydroxycamptothecin, is a biologically active metabolite of irinotecan. Its poor solubility restricted its development as an anticancer agent. We have developed an easy-to-use liposome-entrapped SN-38 (LE SN38) and evaluated its toxicology, pharmacokinetics and antitumor efficacy profile. MATERIALS AND METHODS: Toxicity and pharmacokinetics studies were conducted in CD2F1 mice and beagle dogs. Therapeutic efficacy studies were performed in murine leukemia (P388 and P388/ADR) and in a human pancreatic (Capan 1) tumor models. RESULTS: Multiple dose administration (i.v. x 5) of LE-SN38 indicated a maximum tolerated dose (MTD) of 5.0 and 7.5 mg/kg/day for male and female mice, respectively. The MTD of LE-SN38 in dogs was 1.2 mg/kg. The elimination half-life (t1/2) of SN-38 in mouse plasma was 6.38 h with volume of distribution (VdSS) 2.55 L/kg. In dogs, t1/2 and VdSS were 1.38-6.42 h and 1.69 5.01 L/kg; respectively. P388 tumor-bearing mice dosed with LE-SN38 at 5.5 mg/kg (i.v. x 5) showed 100% survival. LE-SN38 at 4 or 8 mg/kg (i. v. x 5) inhibited 65% and 98% tumor growth, respectively, in a human pancreatic tumor model. CONCLUSION: LE-SN38 showed a favorable pharmacokinetics profile and can be administered safely at therapeutically effective doses. PMID- 15816557 TI - In vivo antitumor activity of folate receptor-targeted liposomal daunorubicin in a murine leukemia model. AB - BACKGROUND: Folate receptor (FR) is selectively amplified among human tumors, including in 70% of myeloid leukemias. FR-targeted liposomal delivery is an attractive strategy for enhancing the therapeutic efficacy of anticancer agents against FR(+) tumors. In this study, FR-targeted liposomal daunorubicin was evaluated in an FR+ L1210JF murine ascites tumor model for therapeutic efficacy in vivo. MATERIALS AND METHODS: FR-targeted liposomal daunorubicin (F-L-DNR) and non-targeted liposomal daunorubicin (L-DNR) were prepared by polycarbonate membrane extrusion followed by remote loading of DNR. FR-targeted liposomal uptake by L1210JF cells was characterized in vitro using fluorescent liposomes entrapping calcein. For in vivo therapeutic study, B6D2F1 mice on a folate-free diet were intraperitoneally implanted with FR (+) L1210JF cells and treated with 4 intraperitoneal injections of 10 mg/kg liposomal DNR at 1, 5, 9 and 13 days following tumor cell inoculation. Animal survival was then monitored daily. RESULTS: LI210JF cells showed approximately 10(3) times greater uptake for FR targeted liposomal calcein compared to the non-targeted control. Uptake of the targeted liposomes could be blocked by 1 mM folic acid. In the therapeutic study, mice treated with F-L-DNR showed significantly greater tumor inhibition and 40.7% greater increase in life-span compared to those that received identical doses of L-DNR. Meanwhile, free DNR given at the same dose failed to prolong the survival of the treated mice. CONCLUSION: F-L-DNR can effectively target FR(+) leukemia cells in vivo. Further preclinical evaluation is warranted to determine its potential application in leukemia therapy. PMID- 15816558 TI - B-cell differentiation, apoptosis and proliferation in diffuse large B-cell lymphomas. AB - Diffuse large B-cell lymphomas (DLBCL) represent the most common type of adult non-Hodgkin's lymphomas in Western countries and are characterized by heterogeneous clinical, histological, immunophenotypic and genetic features. Recent investigations using cDNA and oligonucleotide microarrays have identified molecularly distinct groups of DLBCL with respect to the B-cell differentiation gene expression profile: the germinal center (GC) B-cell-like DLBCL, the activated B-cell-like DLBCL and the type 3 DLBCL. The GC B-cell-like DLBCL were characterized by the expression of genes of the normal GC B-cells, the activated B-cell-like DLBCL were characterized by the expression of genes that are normally induced luring in vitro activation of peripheral blood B-cells, while the type 3 DLBCL did not express either set of genes at a high level. Patients with GC B cell-like DLBCL had more favorable clinical outcome than those with activated B cell-like or type 3 DLBCL. Immunohistochemical studies have shown that the bc16/CD10/MUM1/CD138 B-cell differentiation immunophenotypes are prognostically relevant and may predict the cDNA classification in a sizable fraction of DLBCL. In the last few years, there has been accumulating molecular and immunohistochemical evidence indicating links between B-cell differentiation gene expression profiles and expression of apoptosis and cell cycle-associated genes in DLBCL. The present review summarizes data with respect to the relationships between B-cell differentiation, apoptosis and proliferation in DLBCL. PMID- 15816559 TI - Induction of apoptosis in human promyelocytic leukemia cells by a natural trachylobane diterpene. AB - BACKGROUND: Trachylobane diterpenes are secondary metabolites, quite rare in nature, and their bioactivities are poorly understood. Recently, we have described the cytotoxic activity of ent-trachyloban-3beta-ol isolated from the leaves of Croton zambesicus, a plant used in African folk medicine. MATERIALS AND METHODS: Cell viability on several cell lines, cell morphology, DNA laddering, annexin Vand caspase-3 activation experiments were undertaken in order to analyse the cytotoxicty of trachylobane diterpene and to determine if this compound is able to induce apoptosis. RESULTS: ent-Trachyloban-3beta-ol exerts a dose dependent cytotoxic effect, which varies between cell lines. Induction of apoptosis in HL-60 cells could be detected at a concentration of 50 microM after 24-h treatment. CONCLUSION: We show here, for the first time, that a trachylobane diterpene is able to induce apoptosis in human promyelocytic leukemia cells via caspase-3 activation in a concentration-dependent manner. PMID- 15816560 TI - Caveolin-1 down-regulation activates estrogen receptor alpha expression and leads to 17beta-estradiol-stimulated mammary tumorigenesis. AB - Constitutive activation of estrogen receptor alpha (ER-alpha) expression is an early event in breast cancer tumorigenesis. However, the mechanism whereby ER alpha is constitutively activated during transformation of normal mammary cells has not been well established. Previously, we reported that haploinsufficiency of caveolin-1, a major structural protein that forms caveolae, resulted in anchorage independent growth of a normal mammary epithelial cell line, MCF10A. Here, we further demonstrated that ER-alpha but not ER-beta expression was constitutively activated in these caveolin-1 haploinsufficient cells. Transient treatment of MCF10A cells with beta-methyl-cyclodextrin, a chemical that can displace caveolin 1 from the plasma membrane, also stimulated ER-alpha expression. We further found that the 17beta-estradiol (E2) accelerated anchorage-independent growth of these cells in vitro and promoted their tumorigenesis in nude mice. These results suggest that dysregulation of caveolin-1 is one of the mechanisms by which ER alpha expression is activated during initiation of breast tumorigenesis. PMID- 15816561 TI - Cadherin-6 gene expression in conventional renal cell carcinoma: a useful marker to detect circulating tumor cells. AB - BACKGROUND: Dissemination of cancer cells into the circulation is an essential step in the development of a metastasis. Detection of circulating cancer cells may improve the monitoring methods for cancer patients. However, the detection of circulating renal cancer cells is mainly hampered by the lack of markers available for renal cell carcinoma (RCC). In this study, we evaluated cadherin-6 mRNA as a new molecular marker for the detection of circulating renal cancer cells. MATERIALS AND METHODS: Forty-six blood samples of conventional RCCs were included. A standard protocol of RT-PCR, assisted by computer densitometric analysis to establish a cut-off, was performed to examine cadherin-6 mRNA expression by using specific primers. A renal cancer cell line, SKRC-59 and forty tumor biopsies from conventional RCCs were used as positive controls. Twenty-five blood samples from non-RCC patients were also analyzed. RESULTS: Cadherin-6 mRNA could be detected in 38140 (95%) conventional RCC specimens. Cadherin-6 mRNA was positive in 21/46 (45.7%) blood samples of RCC patients, while no positivity was found in non-RCC blood samples. Among the localized RCCs, 14/35 (40.0%) blood samples were positive while 7/11 (63.6%) were positive among the blood samples from metastatic RCCs. CONCLUSION: Our data indicate that cadherin-6 gene is frequently expressed in conventional RCCs. Cadherin-6 is a useful molecular marker to detect the circulating cancer cells disseminated from conventional RCC. PMID- 15816562 TI - Human sarcoma cell lines MES-SA and MES-SA/Dx5 as a model for multidrug resistance modulators screening. AB - The choice of cell lines for multidrug resistance (MDR) modulators screening may affect the results obtained. Screening is most often performed in model systems which employ cell lines derived from haematological malignancies. Cell lines originating from solid tumours are far less popular. In the present work, we aimed to test the usefulness of the drug-sensitive human sarcoma cell line MES SA, and its multidrug-resistant counterpart MES-SA/Dx5, as a model system for modulators' anti-MDR potency evaluation. Overexpression of P-glycoprotein in the resistant but not in the sensitive cell line was confirmed by flow cytometry and confocal microscopy. Flow cytometry demonstrated that verapamil and trifluoperazine reduced MDR in MES-SA/Dx5 cells as assessed by the rhodamine 123 accumulation test. Both modulators also restored in MES-SA/Dx5 cells the drug accumulation pattern typical for sensitive cells, as judged by confocal microscopy. We conclude that the MES-SA and MES-SA/Dx5 cell line pair constitute a good model for MDR modulators study. PMID- 15816563 TI - Induction of apoptosis by sodium fluorosilicate treatment in human osteogenic sarcoma (HOS) cells. AB - Fluorine compounds are widely used for the prevention of caries, and recently sodium fluorosilicate has been used in water fluorination. The cytotoxic effects of sodium fluorosilicate in several osteosarcoma and oral cancer cells were evaluated in this study by measurement of inhibition of cell proliferation. Human osteogenic sarcoma (HOS) cells were the most sensitive to sodium fluorosilicate treatment. Induction of apoptosis, such as nucleosomal DNA fragmentation and the appearance of apoptotic bodies, were observed in HOS cells by agarose gel electrophoresis and by flow cytometric analysis, respectively. The molecular mechanism of apoptosis induction in HOS was investigated by Western blot analysis. The level of Bcl-2 was decreased and consequent release of cytochrome c was increased. Caspase-3 was activated and the cleavage of poly (ADP-ribosyl) polymerase was increased. In conclusion, sodium fluorosilicate induces apoptosis in HOS cells through decrease in Bcl-2, the release of cytochrome c to the cytosol and activation of caspase-3. PMID- 15816564 TI - Novel D-ring analog of epigallocatechin-3-gallate inhibits tumor growth and VEGF expression in breast carcinoma cells. AB - The cancer chemopreventive activity of green tea and its major polyphenolic constituent, epigallocatechin-3-gallate (EGCG) have been attributed to its antioxidant, antiproliferative and antiangiogenic effects. Several new molecular targets for EGCG's anticarcinogenic activity have been proposed in the recent literature. However, the understanding of the molecular mechanisms of EGCG's activity in vivo have been confounded by its low oral bioavailability and low plasma levels. Studies of EGCG would be greatly aided by the availability of synthetic analogs of EGCG designed to understand the contributions of the A, B, and D-rings and the phenolic hydroxyl groups of EGCG to its molecular mechanisms of action. We recently reported the de novo synthesis of a D-ring analog of EGCG, with the objective of using such analogs to understand the molecular mechanisms of EGCG action. We report here the first studies with a synthetic D-ring analog of EGCG. We examined the ability of the synthetic D-ring analog to inhibit tumor cell proliferation in breast carcinoma cells. We also investigated the effect of the analog on stress-induced VEGF production in breast carcinoma cells using Northern analysis and quantitative RT-PCR. We report here that the synthetic D ring analog inhibits breast cancer cell growth in vitro with potencies equivalent to those of EGCG. Our results also show that, like EGCG, the synthetic analog inhibits hypoxia- and serum starvation-induced production of VEGF mRNA in breast cancer cells. Such synthetic analogs are valuable for understanding the structure function relationship of EGCG and identifying relevant mechanisms of the chemopreventive action of EGCG. PMID- 15816565 TI - Pharmacokinetics and pharmacodynamics of rocuronium in patients with and without renal failure. AB - BACKGROUND AND OBJECTIVE: This study clarifies the relationship between the neuromuscular blocking effects of rocuronium 0.6 mg kg(-1) and its pharmacokinetics in patients with renal failure. METHODS: Seventeen healthy patients and 17 patients with renal failure were studied under propofol anaesthesia in this prospective open label study. Rocuronium 0.6 mg kg(-1) was given after induction of anaesthesia. The train-of-four mechano-myographic response of the thumb to supramaximal stimulation of the ulnar nerve at 2 Hz every 12 s was measured. Venous blood samples (4 mL) were obtained at 0, 2, 4, 7, 10, 15, 20, 30, 60, 120, 180, 240 and 360 min after relaxant administration. Samples were centrifuged, separated and stored at -20 degrees C until plasma levels of rocuronium and its metabolites were measured. Two- and three exponential equations were used to describe the pharmacokinetic data in each group and these were compared to each other using the Wilcoxon signed rank sum test as was the pharmacodynamic data. P < 0.05 was significant. RESULTS: Onset of block was similar in both groups. Clinical duration and the time to recovery of the train-of-four to 70% were prolonged in the renal failure group compared to control; 49 vs. 32 min (P < 0.004, 95% confidential, interval 17, difference 5 28) and 88 vs. 55 min (P < 0.001, 95% confidential interval 33, difference 17 50), respectively. Clearance of rocuronium was reduced by 39% in the renal failure patients compared to control, with an 84% increase in the mean residence time. The volume of distribution was unaffected by renal failure. CONCLUSIONS: The duration of action of a bolus dose of 0.6 mg kg(-1) rocuronium is increased significantly in patients with end-stage renal failure compared to healthy controls. This increase may be due to a decreased clearance of rocuronium, the disease process causing the renal failure and/or the medication which patients with renal failure need in their treatment. PMID- 15816566 TI - Acceleromyography in neonates and small infants: baseline calibration and recovery of the responses after neuromuscular blockade with rocuronium. AB - BACKGROUND: We have evaluated the use of the TOF-Guard (TOF, train-of-four) acceleromyographic thumb responses to ulnar nerve stimulation in neonates and infants and the return of the responses after neuromuscular blockade. METHODS: Baseline acceleromyographic recording of thumb adduction to ulnar nerve stimulation during volatile anaesthesia was performed in 22 babies aged less than 30 weeks. At the start of stimulation the automatic set-up procedure of the TOF Guard was used to see if a 100% control twitch height could be achieved. Irrespective of the ability to achieve a 100% control twitch height, TOF stimulation was used thereafter. When no automatic 100% control twitch could be reached, the transducer signal gain factor was set manually to obtain a 100% value. In 14 of the 22 children the recovery after neuromuscular blockade with rocuronium 0.3 mg kg(-1) was recorded. RESULTS: In nine of 22 patients a 100% baseline twitch height was obtained with the automatic set-up. In the other 13 babies the TOF-Guard display indicated that the transducer signal was too low. The mean time to recovery of control twitch to 75% of baseline after rocuronium 0.3 mg kg(-1) was 51 min (SD = 21) and the time to recovery of the TOF ratio to 70% was 49 min (SD = 19). The mean final twitch height and TOF after recovery from rocuronium blockade were 101% (SD = 15) and 92% (SD = 12), respectively. CONCLUSION: The recovery of the responses after neuromuscular blockade to near baseline values shows that acceleromyography can be used to measure neuromuscular block and recovery in neonates and infants. PMID- 15816567 TI - Cisatracurium, but not mivacurium, induces apoptosis in human umbilical vein endothelial cells in vitro. AB - BACKGROUND AND OBJECTIVE: Cisatracurium is an intermediate acting, non depolarizing neuromuscular blocking agent. Previous reports have indicated a growth-inhibitory effect of the isoforms cisatracurium and atracurium on two human cell lines in vitro. These effects were ascribed to oxidative stress elicited by acrylate esters formed during cisatracurium breakdown. Oxidative stress is a potent precipitator of apoptosis. Therefore, the aim of the present study was to investigate whether the growth-inhibitory effects of cisatracurium could be explained by initiation of apoptosis. METHODS: Human umbilical vein endothelial cells were incubated with cisatracurium at concentrations of 0.96, 3.2, 9.6, 32 and 96 micromol for 24 h. DNA fragmentation was measured using the Cell Death Detection ELISA Plus assay (Roche Diagnostics, Mannheim, Germany). RESULTS: There was a dose dependency of cisatracurium with respect to the rate of apoptosis in human umbilical vein endothelial cells. Programmed cell death could be demonstrated at concentrations encountered in human plasma after single-bolus injections of cisatracurium. Apoptosis was attenuated by the concomitant administration of glutathione. CONCLUSIONS: These findings strongly support the hypothesis that acrylate esters, breakdown products of cisatracurium, induce oxidative stress and, subsequently, apoptosis. PMID- 15816568 TI - Gabexate mesilate hastens recovery from vecuronium-induced neuromuscular blockade. AB - BACKGROUND AND OBJECTIVE: To test the hypothesis that gabexate mesilate, a protease inihibitor, hastens recovery from neuromuscular blockade, we examined the effect of gabexate mesilate on the recovery of vecuronium-induced neuromuscular blockade in anaesthetized patients in a double-blind, randomized fashion. METHODS: Thirty adult patients were divided into two groups of 15. In the gabexate mesilate group, immediately after administration of vecuronium 0.1 mg kg(-1), a continuous infusion of gabexate mesilate was started at a speed of 1.5 mg kg(-1) h(-1). In the control group, normal saline was administered instead of gabexate mesilate. Times to the return of T1, T2, T3 or T4 (first, second, third and fourth response of train-of-four (TOF)), times to the recovery of T1/control to 0.25 (T25) or 0.5 (T50), recovery of T1/control or TOF ratio (T4/T1) were compared between the two groups. RESULTS: Times to the returns of T1, T2, T3 and T4 in the gabexate mesilate group were significantly shorter than in the control group (19.4 +/- 6.8 vs. 25.7 +/- 7.2 min for T1; mean +/- SD, P = 0.020). Times to T25 and T50 were significantly shorter in the gabexate mesilate group than in the control group (34.0 +/- 9.9 vs. 51.3 +/- 10.2 min for T25, P < 0.001). T1/control and TOF ratio in the gabexate mesilate group were significantly higher than in the control group 40-80 min and 40-120 min after administration of vecuronium, respectively (P < 0.05). CONCLUSION: Gabexate mesilate hastens recovery from neuromuscular block in anaesthetized patients receiving vecuronium. PMID- 15816569 TI - Large bolus dose vs. continuous infusion of cisatracurium during hypothermic cardiopulmonary bypass surgery. AB - BACKGROUND AND OBJECTIVE: We investigated whether a high bolus dose of cisatracurium (8x ED95) given at induction can provide muscle relaxation for the major part of a cardiac procedure with hypothermic cardiopulmonary bypass, avoid important postoperative residual curarization and cause no waste of product. METHODS: Twenty patients were randomly assigned either to Group 1 (n = 10) or Group 2 (n = 10). Those in Group 1 were given cisatracurium in a high bolus dose (0.4 mg kg(-1)). Those in Group 2 received cisatracurium 0.1 mg kg(-1) at induction followed after 30 min by a continuous infusion of cisatracurium. As an escape medication in case of patient movement, a bolus dose of cisatracurium 0.03 mg kg(-1) was given. RESULTS: In Group 1 (large cisatracurium bolus dose), the clinical duration of effect (until T1/T0 = 25%) was 110 min. Six of 10 patients in Group 1 required additional boluses of cisatracurium intraoperatively. Four of these six had received an additional bolus near the end of surgery and had a train-of-four (TOF) ratio = 0 at the end. The other four patients in Group 1 had a final TOF ratio >0.9. In Group 2 (continuous cisatracurium infusion), only two patients had a TOF ratio >0.9 at the end of surgery, no patient moved and none received additional boluses. The total amount of cisatracurium used in the bolus and infusion Groups was 34.5 +/- 7.8 and 21.3 +/- 5.7 mg, respectively (P = 0.0004). CONCLUSIONS: For continued neuromuscular block during hypothermic cardiac surgery, a high bolus dose of cisatracurium appears to be safe, although it is not an alternative to a continuous infusion, as its neuromuscular blockade does not cover the intraoperative period and a high incidence of movements occurs. In the patients who received a high bolus dose of cisatracurium, postoperative residual curarization appeared after additional boluses had been given. The consumption of cisatracurium by high bolus was significantly greater than with continuous infusion. PMID- 15816570 TI - Comparison of three modes of patient-controlled epidural analgesia during labour. AB - BACKGROUND AND OBJECTIVES: This study compares three modes of patient-controlled epidural analgesia in parturients during labour. METHODS: Eighty-four women were randomized to one of three groups. The epidural solution used in all the three groups was 0.1% bupivacaine with fentanyl 2 microg mL(-1). Patients were able to self administer a demand dose of 3 mL with a lockout interval of 6 min in Group A, 6 mL with a lockout interval of 12 min in Group B and 9 mL with lockout interval of 18 min in Group C. All patients received a background infusion at a rate of 6 mL h(-1). Visual analogue pain scores, pinprick analgesia and motor block were assessed hourly by a blinded observer. The physician-administered supplementation and the cumulative dose of bupivacaine were also compared between the three groups. RESULTS: Pain scores, sensory level and motor block were not different among the study groups. Patients' satisfaction was rated good to excellent with no difference among groups. The cumulative dose of bupivacaine was not significantly different. However, there was a trend towards a decreased need for rescue analgesia in Group C. Within each group, the physician-administered supplementations were significantly higher during the second stage of labour than during the first stage (P < 0.05). CONCLUSION: The three modes of patient controlled epidural analgesia supplemented by a background infusion of 6 mL h(-1) were equally effective for labour analgesia with a trend for decreased rescue analgesia in the group with a larger bolus dose and a longer lockout interval. PMID- 15816571 TI - Transient neurological symptoms with subarachnoid lidocaine: effect of early mobilization. AB - BACKGROUND AND OBJECTIVE: Although lidocaine has been used extensively for spinal anaesthesia since 1949, it has been associated with transient neurological symptoms only in the past 10 yr. It has been suggested that early ambulation after spinal anaesthesia, as opposed to traditional 24 h recumbency, might be the causative factor for the development of transient neurological symptoms. The purpose of this study was to examine the effect of early ambulation on the incidence of transient neurological symptoms after single injection spinal anaesthesia with lidocaine 2%. METHODS: Sixty patients undergoing minor surgery under spinal anaesthesia were included. All patients received lidocaine 60 mg. After the establishment of successful subarachnoid block, patients were randomly allocated to two groups of 30 patients. Patients in Group 1 were ambulated as soon as possible, whereas patients in Group 2 were kept recumbent until 6 h after subarachnoid injection. Two days after surgery patients were contacted by a blinded observer and interviewed of transient neurological symptoms using a standardized questionnaire. Patients were asked to express the intensity of pain/discomfort on a verbal rating scale from 0 (no pain) to 10 (worst pain imaginable). RESULTS: There was no significant difference in the incidence of transient neurological symptoms (23% vs. 27%). In all patients, symptoms resolved completely within 6-24 h. The median pain score was 5 (range 2-8) in Group 1 and 7 (range 1-8) in Group 2. CONCLUSIONS: Under the conditions of this study, there is no correlation between the time of ambulation after spinal anaesthesia with lidocaine and the incidence of transient neurological symptoms. PMID- 15816572 TI - Remifentanil for analgesia during retrobulbar nerve block placement. AB - BACKGROUND AND OBJECTIVES: Patients undergoing eye surgery under regional anaesthesia often require concomitant medication for analgesia and comfort. Remifentanil, with its ultra-short acting-profile, may be useful to reduce pain during retrobulbar nerve block for cataract surgery. METHODS: We performed a prospective, randomized, double-blind study to compare the efficacy of remifentanil for analgesia during retrobulbar nerve block placement. Ninety patients undergoing cataract surgery were randomly divided to receive either remifentanil 0.3 microg kg(-1) (n = 45) or an equivalent volume of saline (n = 45). The injection was administered within 30 s in both groups. Patients rated their amount of pain on a 10 cm visual analogue scale. Respiratory frequency, oxygen saturation, cardiac rhythm and postoperative nausea and vomiting (PONV) were recorded. RESULTS: The mean visual analogue score in the Remifentanil group was 2.56; it was 5.51 in the Saline group (P = 0.001, U-test). Three patients developed bradycardia and three had PONV in the Remifentanil group. Two patients developed tachycardia and one had PONV in the Saline group. No patient developed respiratory depression. CONCLUSION: In patients undergoing retrobulbar block placement for eye surgery, 0.3 microg kg(-1) remifentanil over 30 s significantly reduced their reported pain. In addition, remifentanil did not increase the risk of untoward side-effects. PMID- 15816573 TI - Comparison of ephedrine and ketamine in prevention of injection pain and hypotension due to propofol induction. AB - BACKGROUND AND OBJECTIVE: This prospective, double-blind, randomized, placebo controlled study compares the effects of ephedrine and ketamine on injection pain, and hypotension from propofol. METHODS: After obtaining the approval of the Ethics Committee, 75 patients (ASA I-II) scheduled for elective operations with general anaesthesia were divided into three groups. Saline 2 mL (Group S, n = 25), ketamine 0.5 mg kg(-1) (Group K, n = 25) or ephedrine 70 microg kg(-1) (Group E, n = 25) were administered over 5 s after tourniquet application. After releasing the tourniquet, propofol 2 mg kg(-1) was injected in 30 s. Pain was evaluated on a numerical scale (0-10) where 0 represented no pain and 10 the most severe pain possible. Systolic, diastolic blood pressures and heart rates were recorded preoperatively, 1 min after propofol injection, before intubation and 1, 2 and 3 min after intubation in all patients. RESULTS: The incidences of pain in Groups S, E and K were similar (84%, 80% and 72%, respectively). The mean pain score in Group K (2.1, SD 3.1) was significantly lower than those of Groups S and E (4.9, SD 2.6 and 4.6, SD, 3.3, respectively) (P < 0.05). The systolic and diastolic blood pressure values in Group K (120 +/- 27 mmHg) and Group E (123 +/- 21 mmHg) before intubation were significantly higher than that of Group S (104 +/ 25 mmHg) (P < 0.05). There was no significant difference between the mean heart rate values of the groups. CONCLUSIONS: Low dose ketamine or ephedrine pretreatment may prevent hypotension due to propofol induction. Despite the reduction in injection pain intensity after ketamine, the study drugs were found to be ineffective in lowering the injection pain incidence. PMID- 15816574 TI - Ischaemic preconditioning but not isoflurane prevents post-ischaemic production of hydroxyl radicals in a canine model of ischaemia-reperfusion. AB - BACKGROUND AND OBJECTIVE: Isoflurane has been shown to mimic ischaemic preconditioning (IPC). The protective effect of IPC, or applying isoflurane or perfusion with the 'push-pull' complex zinc-desferrioxamine (Zn-DFO) in the canine heart, was investigated. METHODS: Thirty minutes after salicylate administration (100 mg kg(-1)) the heart was exposed. All dogs were subjected to a 10 min left anterior descending artery occlusion followed by 2 h of reperfusion. In Group I (n = 9) isoflurane (2.5%) was administered 10 min prior to and during ischaemia. In Group II (n = 8), IPC was elicited by 5 min coronary artery occlusion, followed by 5 min of reperfusion, prior to the 10 min ischaemia. In Group III (n = 9) Zn-DFO (2.5 mg kg(-1)) was given 10 min prior to ischaemia. The effects of these interventions were compared to control (n = 10). Coronary sinus blood concentrations of salicylate, 2,3-dihydroxybenzoic acid (DHBA), lactate, pH and oxygen content were monitored. RESULTS: In the control group, 2,3-DHBA increased by 32% above the pre-ischaemic value (P < 0.05). In contrast, in the IPC hearts, a significant decrease in the production of 2,3-DHBA was observed (40% lower than baseline, P < 0.01). In the isoflurane group only a 13% (and non-significant) decrease was noticed. In the Zn-DFO group a 33% decrease was found (P < 0.01). The increase in lactate concentrations in the IPC and Zn-DFO groups was significantly smaller than that of control and isoflurane groups. CONCLUSIONS: IPC protected the heart against the deleterious effects of reperfusion, possibly by amelioration of the level of oxygen-derived reactive species, and the complete inhibition of reactive hydroxyl radical production. Isoflurane did not prove to be as effective in reducing the free radical damage. PMID- 15816575 TI - Target-controlled infusion of remifentanil or fentanyl during extra-corporeal shock-wave lithotripsy. AB - BACKGROUND AND OBJECTIVE: Target-controlled infusions (TCIs) of remifentanil and fentanyl in conscious sedation regimes for extra-corporeal shock-wave lithotripsy have not been reported. We estimated the effect site concentrations of remifentanil and fentanyl needed to obtain adequate analgesia in 50% of patients (EC50) and compared both drugs in terms of intra- and post-procedure complications. METHODS: Forty-four adult patients were randomly distributed into two groups: Group R received remifentanil and Group F received fentanyl TCI with initial effect site concentrations of 1.5 and 2 ng mL(-1), respectively. Pain was assessed using a 10-point verbal analogue scale and <3 was considered adequate analgesia. Increments or decrements of 0.5 ng mL(-1) were then introduced for subsequent patients according to Dixon's up and down method. During the rest of the procedure, TCI was adjusted to maintain verbal analogue scale <3. RESULTS: Remifentanil and fentanyl EC50 were 2.8 ng mL(-1) (95% confidence interval (CI): 1.8-3.7 ng mL(-1)) and 2.9 ng mL(-1) (95% CI: 1.7-4.1 ng mL(-1)), respectively (n.s.). At EC50, the probability of having a respiratory rate <10 was 4% (95% CI: 0-57%) for remifentanil and 56% (95% CI: 13-92%) for fentanyl. Hypoxaemia, vomiting and sedation were more frequent in Group F during and after the procedure (P < 0.05). CONCLUSIONS: A similar EC50 but more respiratory depression, sedation and PONV were found with fentanyl TCI than with remifentanil TCI. PMID- 15816576 TI - Arginine vasopressin reduces cerebral oxygenation and cerebral blood volume during intact circulation in swine---a near infrared spectroscopy study. AB - BACKGROUND AND OBJECTIVE: The aim of the present study was to investigate the impact of arginine vasopressin (AVP), a drug currently under investigation for use during cardiopulmonary resuscitation, on cerebral oxygenation and cerebral blood volume (CBV) in pigs with intact systemic circulation using near infrared spectroscopy. METHODS: Nine healthy pigs were anaesthetized and subjected to invasive haemodynamic monitoring as well as to non-invasive determination (with near infrared spectroscopy) of changes in the Tissue Oxygenation Index (TOI is the ratio of oxygenated to total tissue haemoglobin), Tissue Haemoglobin Index (THI, representing CBV) and cytochrome oxidase (deltaCytOx, representing the balance of intracellular oxygen supply). RESULTS: At both 3 and 5 min after AVP administration, TOI, THI and deltaCytOx were significantly (P < 0.001) reduced compared to baseline, while cerebral perfusion pressure increased significantly (P < 0.001). The effects of AVP on TOI and THI lasted longer than on deltaCytOx. There were no significant changes with respect to the intracranial pressure throughout the study period. CONCLUSIONS: No improvement of cerebral oxygenation was detected after AVP administration in swine with an intact systemic circulation. In contrast to recently published investigations, AVP provoked a sustained drop in indices of cerebral oxygenation and CBV. PMID- 15816577 TI - Low-dose ketamine with clonidine and midazolam for adult day care surgery. PMID- 15816578 TI - Ketamine pretreatment with venous occlusion attenuates pain on injection with propofol. PMID- 15816579 TI - Influence of an external pacemaker on bispectral index. PMID- 15816580 TI - Anaphylactic reaction after rocuronium. PMID- 15816581 TI - Complete recovery from prolonged cardiac arrest following self-administration of cisatracurium. PMID- 15816582 TI - A clinical comparison between bispectral index (BIS) and high frequency EEG signal detection (SNAP). PMID- 15816583 TI - The laryngeal mask airway (LMA) in paediatric ophthalmic anaesthesia practice. PMID- 15816584 TI - Thoracic epidural analgesia for coronary artery surgery. A bridge too far? PMID- 15816585 TI - Serum creatinine and perioperative outcome--a matched-pairs approach using computerised anaesthesia records. AB - BACKGROUND AND OBJECTIVE: A study was designed to utilise the resources of our computerised anaesthesia record keeping system to assess the attributable effects of increased preoperative creatinine (> 1.3 mg dL(-1)) on outcome in patients undergoing non-cardiac surgery. METHODS: This retrospective study was based on data sets of 58 458 patients recorded with a computerised anaesthesia record keeping system over a period of 4 yr at a tertiary care university hospital. Cases were defined as patients with a preoperative creatinine > 1.3 mg dL(-1); controls (creatinine < or = 1.3 mg dL(-1)) were selected and automatically matched according to several parameters (ASA physical status, high risk and urgency of surgery, age and gender) in a stepwise fashion. Main outcome measures were hospital mortality and the incidence of intraoperative cardiovascular events. RESULTS: Three-thousand-and-twenty-eight patients (5.2%) had preoperative creatinine values > 1.3 mg dL(-1). Matching was successful for 54.5% of the cases, leading to 1649 cases (mean creatinine 3.3 +/- 2.2 mg dL(-1)) and 1649 controls (1.0 +/- 0.2 mg dL(-1)). The crude mortality rates for the cases and matched controls were 2.2% (n = 36) and 0.9% (n = 15), respectively (P = 0.003). Intraoperative cardiovascular events were found in 30.1% of the patients (n = 496) and in 28.3% of the matched controls (n = 466; P = 0.25, power = 0.46). Using logistic regression analyses a significant association between preoperative increased creatinine and hospital mortality was found (odds ratio 2.62; 95% confidence interval [1.39; 4.931). CONCLUSIONS: An increased preoperative serum creatinine in patients undergoing non-cardiac surgery is associated with an increased perioperative risk, but not with a higher incidence of intraoperative cardiovascular events. PMID- 15816586 TI - Stress reduction and analgesia in patients exposed to calming music postoperatively: a randomized controlled trial. AB - BACKGROUND AND OBJECTIVES: This randomized controlled trial was designed to evaluate, first, whether intra- or postoperative music therapy could influence stress and immune response during and after general anaesthesia and second, if there was a different response between patients exposed to music intra- or postoperatively. METHOD: Seventy-five patients undergoing open hernia repair as day care surgery were randomly allocated to three groups: intraoperative music, postoperative music and silence (control group). Anaesthesia and postoperative analgesia were standardized and the same surgeon performed all the operations. Stress response was assessed during and after surgery by determining the plasma cortisol and blood glucose levels. Immune function was evaluated by studying immunoglobulin A (IgA) levels. Patients' postoperative pain, anxiety, blood pressure (BP), heart rate (HR) and oxygen saturation were also studied as stress markers. RESULTS: There was a significantly greater decrease in the level of cortisol in the postoperative music group vs. the control group (206 and 72 mmol L(-1) decreases, respectively) after 2 h in the post anaesthesia care unit. The postoperative music group had less anxiety and pain and required less morphine after 1 h compared with the control group. In the postoperative music group the total requirement of morphine was significantly lower than in the control group. The intraoperative music group reported less pain after 1 h in the post anaesthesia care unit. There was no difference in IgA, blood glucose, BP, HR and oxygen saturation between the groups. CONCLUSION: This study suggests that intraoperative music may decrease postoperative pain, and that postoperative music therapy may reduce anxiety, pain and morphine consumption. PMID- 15816587 TI - Influence of posture on the incidence of vein cannulation during epidural catheter placement. AB - BACKGROUND AND OBJECTIVE: Epidural vein cannulation has long been recognized as a problem in parturients due to distension of epidural veins. Epidural vein engorgement is maximal when the pregnant woman is in the supine position and minimal in the lateral position. Following an initial observation of an apparently high incidence of epidural vein cannulation in the sitting position, a randomized trial was conducted to document whether such an association existed. METHODS: A total of 209 term parturients were randomized to either the sitting or lateral position (107 left lateral, 102 sitting). Epidural catheter placement was achieved using a loss of resistance to air technique with an 18-G Tuohy needle. A data sheet was completed for each patient recording patient position, patient characteristics data, stage in labour and the incidence of epidural vein cannulation. Poor labour analgesia was defined as a visual analogue scale (VAS) > 40 mm on a 0-100 mm pain intensity VAS. RESULTS: The risk of epidural vein cannulation was significantly higher in the sitting group (16 of 102 = 15.7%) compared with the lateral position group (4 of 107 = 3.7%), P = 0.011. There was a significant association between epidural vein cannulation and poor analgesia (P = 0.006). These two variables remained independently significant on multiple regression analysis (position, P = 0.009; analgesia, P = 0.006). CONCLUSIONS: We conclude that there is a direct correlation between the incidence of epidural vein cannulation and patient posture during epidural catheter insertion in parturients. PMID- 15816588 TI - Is there a place for interscalene block performed after induction of general anaesthesia? AB - BACKGROUND AND OBJECTIVE: The timing of interscalene block in relation to general anaesthesia remains a controversial subject. We believe that the results of our study demonstrate that this block may be performed safely on anaesthetized patients, providing that certain conditions are met. METHODS: Five hundred and forty-eight cases of arthroscopic shoulder surgery were analysed retrospectively in order to identify the results of interscalene block performed after induction of general anaesthesia, but before the start of surgery. Patients were kept in hospital for 24 h and assessed before being discharged from the hospital and again in the period from 4 to 8 weeks after surgery. Duration of the block, quality of postoperative pain relief and neurological complications were recorded. RESULTS: Our study failed to reveal any permanent or long-term neurological complications attributable to interscalene block performed after induction of general anaesthesia. CONCLUSION: It is our opinion that the primary factor for safe interscalene block is modification of the anaesthetic technique rather than the timing of regional block in relation to induction of general anaesthesia. PMID- 15816589 TI - Spinal anaesthesia in full-term infants of 0-6 months: are there any differences regarding age? AB - BACKGROUND AND OBJECTIVE: The aim of the study was to report our experience concerning the effectiveness, complications and safety of spinal anaesthesia, and to determine whether spinal anaesthesia was effective in full-term infants undergoing elective inguinal hernia repair. METHODS: Sixty-eight full-term infants aged < 6 months were included in the study. Infants were divided into three groups; Group I (< 1 month, n = 20), Group II (> 1 and < 3 months, n = 26), and Group III (3-6 months, n = 22). All spinal blocks were performed under mask inhalation anaesthesia. A dose of bupivacaine 0.5% 0.5 mg kg(-1) was used for infants under 5kg and 0.4 mg kg(-1) for those over 5 kg. Heart rate, mean arterial pressure, respiratory rate and SPO2 were recorded before and after spinal anaesthesia at 5 min intervals. Time to onset of analgesia, time to start of operation, duration of operation, anaesthesia and hospitalization, postoperative analgesic requirement and complications were recorded. RESULTS: Adequate spinal anaesthesia without sedation was better, time to obtain maximum cutaneous analgesia was shorter and need for sedation and postoperative analgesic requirement were significantly lower in Group I. Although heart rate, mean arterial pressure and respiratory rate decreased < 20% in all groups following spinal analgesia, the decrease in Group I was lower than the others. CONCLUSIONS: Spinal anaesthesia is an effective choice in inguinal hernia repair for full-term infants aged < 1 month, providing excellent and reliable surgical conditions. However, this technique is not as useful for infants aged between 1 and 6 months. PMID- 15816590 TI - The ProSeal laryngeal mask airway and the laryngeal tube Suction for ventilation in gynaecological patients undergoing laparoscopic surgery. AB - BACKGROUND AND OBJECTIVE: ProSeal Laryngeal Mask Airway (PLMA) and Laryngeal Tube Suction (LTS), supraglottic airway devices allowing gastric drainage, were compared in this prospective, randomized study for airway management under conditions with elevated intra-abdominal pressure induced by capnoperitoneum. METHODS: Fifty patients undergoing elective gynaecological laparoscopic surgery were randomized to two groups of 25 each. After induction of general anaesthesia, devices were inserted, correct placement was verified, airway leak pressure was measured, and a gastric tube was inserted. Ease of insertion, quality of airway seal, risk of gastric insufflation and patient comfort were investigated. RESULTS: There were no differences in patient characteristics data for both groups. First-time insertion success rates were comparable for both groups: 92%- first attempt, 8%--second attempt for PLMA and LTS. Time until delivery of the first tidal volume for PLMA and LTS was 23.2 +/- 6.1 and 23.5 +/- 6.6s, airway leak pressure was 45.4 +/- 4.9 cmH2O and 45.6 +/- 6.7 cmH2O with cuff pressures adjusted to 60 cmH2O. No gastric insufflation, gas loss or signs of regurgitation were detected. Placement of a gastric tube was successful in all patients. Patients were questioned for sore throat and dysphagia after removal of devices. Sore throat was stated in 1%/0% (PLMA) and 8%/4% (LTS) after 6/24 h, dysphagia in 4%/4% (PLMA) and 12%/4% (LTS). CONCLUSIONS: Both devices provide a secure airway even under conditions of elevated intra-abdominal pressure. In this pilot study, no differences concerning handling or quality of airway seal were detected between PLMA and LTS. PMID- 15816591 TI - Impact of patient age on propofol consumption during propofol-remifentanil anaesthesia. AB - BACKGROUND: This study was designed to investigate the impact of patient age on propofol consumption and recovery time using a propofol-remifentanil anaesthetic standardized with Narcotrend EEG monitoring. The Narcotrend is a monitor for measuring the depth of anaesthesia based upon a six-letter classification from A (awake) to F (increasing burst suppression) including 14 substages. METHODS: In 200 patients scheduled for minor orthopaedic surgery Narcotrend EEG electrodes were positioned on the patient's forehead as recommended by the manufacturer. Anaesthesia was induced with remifentanil 0.4 microg kg(-1) min(-1) and 2 mg kg( 1) propofol. Immediately after intubation remifentanil was reduced to a constant rate of 0.2 microg kg(-1) min(-1) whereas a propofol infusion was now started at 3 mg kg(-1) h(-1) and then adjusted accordingly to achieve a target Narcotrend stages of D(0-2) indicating general anaesthesia. At the end of surgery the propofol and remifentanil infusions were stopped without tapering, the time to unstimulated opening of eyes was determined, and the propofol consumption (given as mg kg(-1) h(-1)) was calculated from the total amount of infused propofol but without the induction bolus, from the actual body weight and the duration of propofol infusion. Furthermore, a linear regression analysis was applied for propofol consumption vs. age. RESULTS: The ages of the patients studied ranged from 16 to 83 yr old and patients were classified as ASA I-III. Propofol consumption significantly decreased with the patients' age: 30 yr of age or below the propofol consumption was calculated as 5.9 +/- 1.7 mg kg(-1) h(-1), for 31-50 yr as 5.4 +/- 1.8 mg kg(-1) h(-1), for 51-70 yr as 4.5 +/- 1.7 mg kg(-1) h(-1) and above 70 yr as 3.5 +/- 1.4 mg kg(-1) h(-1). Linear regression analysis revealed propofol (mg kg(-1) h(-1)) = 9.136 - (0.0597 x age (yr)); R = 0.53. Concomitantly, the recovery time to opening of eyes increased with the patients' age: < or = 30 yr, 7.4 +/- 3.7 min; 31-50 yr, 9.5 +/- 4.0 min; 51-70 yr, 9.8 +/- 4.1 min; and > or = 71 yr, 14.9 +/- 12.1 min. CONCLUSIONS: We conclude that with Narcotrend guidance, mean propofol consumption and recovery times are age dependent. However, as a result of large inter-individual variability, age per se does not allow a prediction of individual propofol need or recovery time. PMID- 15816592 TI - Comparison of cardiac output measurements by arterial trans-cardiopulmonary and pulmonary arterial thermodilution with direct Fick in septic shock. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to compare cardiac output (CO) measurements by arterial trans-cardiopulmonary thermodilution (ATD) and pulmonary arterial thermodilution (PATD) with CO estimated on the basis of the Fick calculation via a metabolic monitor in septic shock. METHODS: In a prospective animal study 20 anaesthetized, ventilated pigs (20.9 +/- 1.9 kg) were investigated. Septic shock was induced with faecal peritonitis. A pulmonary artery catheter was used for conventional measurement of CO. Simultaneously ATD was measured with a thermistor tipped catheter inserted into right carotid artery. Whole body oxygen consumption was measured by indirect calorimetry. Eighty data pairs of simultaneous CO measurements were analysed. RESULTS: CO measured with Fick and that measured with PATD were significantly correlated (r = 0.94, r = 0.87, P < 0.001). Mean CO measured by PATD was 94.3 +/- 40.1 mL min(-1) kg(-1). Bias was 10.1 mL min(-1) kg(-1) (95% confidence interval (CI): 6.0-14.2 mL min(-1) kg(-1)) with limits of agreement of -26.8 to 47.0 mL min(-1) kg(-1). Correlation between Fick derived CO estimation and ATD CO was similar (r = 0.91, r2 = 0.83, P < 0.001). Mean CO measured by trans-cardiopulmonary thermodilution was 104.3 +/- 43.2 mL min(-1) kg(-1). Bias was 0.75 mL min(-1) kg(-1) (95% CI: 3.8 to 5.3 mL min(-1) kg(-1)) with limits of agreement of -39.7 to 41.2 mL min( 1) kg(-1). CONCLUSIONS: Even during haemodynamic instability in septic shock the correlation of arterial trans-cardiopulmonary thermodilution and PATD derived CO with direct Fick was good. As arterial trans-cardiopulmonary thermodilution is less invasive than PATD, the former may offer practical advantages. PMID- 15816593 TI - Mechanisms involved in the relaxing effect of midazolam on coronary arteries. AB - BACKGROUND AND OBJECTIVE: Hypotension, especially in elderly and hypovolaemic patients, is frequently associated with intravenous midazolam administration. The mechanisms are not completely understood. This study was designed to investigate the mechanisms involved in the relaxing effect of midazolam on coronary arteries. METHODS: The substance was studied in isolated porcine coronary artery rings precontracted by either potassium chloride or prostaglandin F2alpha. RESULTS: Midazolam caused vasodilatation in a concentration-dependent manner. Relaxation was more pronounced in prostaglandin F2alpha precontracted segments than in those treated with potassium chloride (P < 0.001). Vasodilatation was unaffected by Nomega-nitro-L-arginine, indomethacin and glibenclamide. Tetraethylammonium chloride, an inhibitor of the BK(Ca) K+ channel (a high conductance Ca(2+) sensitive K+ channel), dose dependently attenuated the vasodilating effect of midazolam (P < 0.01). CONCLUSIONS: Hyperpolarization of the smooth muscle cell in the vessel wall, elicited by the activation the BK(Ca) K+ channel, may contribute to the vasorelaxing effect of midazolam. PMID- 15816594 TI - Moderate hypothermia, but not calpain inhibitor 2, attenuates the proteolysis of microtubule-associated protein 2 in the hippocampus following traumatic brain injury in rats. AB - BACKGROUND AND OBJECTIVE: The degradation of the cytoskeletal protein microtubule associated protein 2 (MAP2), by calpain has been known to occur following traumatic brain injury. We examined the therapeutic potential of calpain inhibitor 2, compared with that of moderate hypothermia in traumatic brain injury produced by weight drop in rats. METHODS: An inhibitor treated group (n = 8) received calpain inhibitor 2 intravenously (i.v.) for 5 min before and for 6 h after injury (total 2 micromol); a hypothermic (HT) group (n = 8) was maintained at 30 degrees C (temporalis muscle temperature) for 45 min prior to and 60 min after injury; an untreated (UT) group (n = 8) received an infusion of inactive vehicle. Eight rats (sham group) underwent surgery without brain injury. Histopathological (haematoxylin and eosin staining) and MAP2 (immunohistchemistry and western blotting) evaluations were performed at 6 h after injury. RESULTS: Ipsilateral cortical damage was marked in the injured groups. In the hippocampus, marked pyramidal neuronal damage was observed in the UT and calpain inhibitor treated (CI) groups, while these neurons were better preserved in the HT group. The hippocampal MAP2 levels in the UT, CI and HT groups were significantly decreased to 13 +/- 9%, 28 +/- 33% and 62 +/- 25% of the sham contol, respectively. MAP2 concentration in the HT group was significantly higher than in UT and CI groups (P < 0.05). CONCLUSION: The results suggest that moderate hypothermia, but not calpain inhibitor 2 with the tested regime, attenuates cytoskeletal damage in the ipsilateral hippocampus at 6 h after traumatic brain injury. PMID- 15816595 TI - Do intracellular, extracellular or urinary magnesium concentrations predict renal retention of magnesium in critically ill patients? AB - BACKGROUND AND OBJECTIVE: Magnesium disorders are common in hospitalized patients. In patients with low or normal magnesium, the intravenous magnesium loading test has been demonstrated to be a sensitive test to assess magnesium deficiency in critically ill patients. However, it is more time consuming and more difficult than the measurement of intracellular or extracellular magnesium concentrations. This study evaluated whether erythrocyte, plasma and urinary magnesium concentrations predict renal magnesium retention measured by th magnesium loading test. METHODS: One-hundred-and-three intensive care patients (36 females, 67 males) in a tertiary care centre and 41 healthy subjects (13 females, 28 males) took part in this prospective study. Intracellular, total plasma, ionize extracellular and urinary magnesium concentrations were measured and also magnesium retention by intravenous magnesium loading test. RESULTS: Total plasma magnesium concentration was poorly correlated with magnesium retention (r = 0.36 r2 = 0.13) and was the only parameter that significantly predicted magnesium retention in intensive care patients (P < 0.01). However, only 10% of the magnesium retention data were linked to the total plasma magnesium concentration. CONCLUSIONS: Total plasma magnesium concentration predicts magnesium retention in critically ill intensive care patients but not intracellular and urinary magnesium concentrations. Only a small proportion of the magnesium retention was due to the total plasma magnesium concentration. PMID- 15816596 TI - Misinterpretation of the TEI-index. PMID- 15816597 TI - Combination of sildenafil and bosentan for nitric oxide withdrawal. PMID- 15816598 TI - The effect of hypotensive anaesthesia on cognitive functions and recovery at endoscopic sinus surgery. PMID- 15816599 TI - Possible underestimation of histamine releasing potency of cisatracurium? PMID- 15816600 TI - Response to cisatracurium in patient with Charcot-Marie-Tooth disease. PMID- 15816601 TI - Costs of anaesthesia. PMID- 15816602 TI - Plasma von Willebrand factor antigen levels in non-small cell lung cancer patients. AB - BACKGROUND: To analyze the behavior of circulating von Willebrand factor antigen (vWf:Ag) in patients with non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Pre-surgical vWf:Ag levels were measured in 64 patients with histologically diagnosed NSCLC compared to 64 patients with benign pulmonary diseases, as well as 64 age- and sex-matched controls. RESULTS: Patients with NSCLC had mean vWf:Ag concentrations lower than either controls or benign patients (p =0.001). CEA was the only variable predictive of low vWf:Ag levels (p<0.01). Five of the 64 NSCLC patients had abnormally low vWf:Ag concentrations (<36 IU/dL). When these patients were excluded from the analysis, the vWf:Ag levels of NSCLC patients did not differ from those of controls (p=0.19). CONCLUSION: The vWf antigen levels of NSCLC patients are not substantially altered. A small subset of these patients will have a depletion of circulating vWf:Ag, probably because of a paraneoplastic process associated with an advanced stage of disease. PMID- 15816603 TI - Paclitaxel plus gemcitabine in advanced non-small cell lung cancer patients with low performance status. AB - BACKGROUND: The aim was to determine the efficacy and safety of a platinum-free regimen combining gemcitabine and paclitaxel for the treatment of patients with advanced non-small cell lung cancer (NSCLC) and a low performance status (PS). PATIENTS AND METHODS: Patients with histologically confirmed unresectable NSCLC, no previous chemotherapy, measurable lesion and a PS of 2 or 3 according to the Eastern Cooperative Oncology Group (ECOG) scale were elegible. Chemotherapy consisted of paclitaxel 200 mg/m2 on day 1 plus gemcitabine 1000 mg/m2 on days 1 and 8, every 3 weeks, for a maximum of 8 cycles. RESULTS: Twenty-nine consecutive patients were enrolled. PS was 2 and 3 in 93% and 7% of patients, respectively. A total of 149 courses of chemotherapy were delivered (median 4.6). Responses: complete response 1 (3.4%), partial response 11 (37.9%), stable disease 12 (41.3%), progressive disease 5 (17.2%) (response rate 41.3%, 95% CI. 23.5% to 61.6%). Median time to progression was 8.3 months (range 2.9-31.7); median overall survival was 13.6 months (range 3.2-31.7). Grade 3 leukopenia occurred in 3% of patients, while grade 3 thrombocytopenia was observed in 25% of patients. CONCLUSION: Reasonable response rates and a satisfactory clinical benefit can be obtained with a platinum-free regimen in NSCLC patients with a low PS. PMID- 15816604 TI - Sudden onset of interstitial lung disease induced by gefitinib in a lung cancer patient with multiple drug allergy. AB - Gefitinib is an oral selective inhibitor of the epidermal growth factor receptor tyrosine kinase which is effective for patients with advanced non-small cell lung cancer. A 75-year-old man with advanced adenocarcinoma of the lung was treated with gefitinib. He had a history of allergy to several antibiotics and Welder's lung. Two days after initiation, he developed acute interstitial lung disease (ILD) and died of respiratory failure due to progression of ILD. Critical assessment pointed to gefitinib as the likely cause of this complication. This is the first report of rapid gefitinib-induced ILD. This case should alert physicians to the potential for dangerous pulmonary side-effects of gefitinib therapy, especially in patients with drug allergy. PMID- 15816605 TI - The influence of surgical volume on hospital mortality and 5-year survival for carcinoma of the oesophagus and gastric cardia. AB - OBJECTIVE: To study the effects of hospital operation volume on hospital mortality and 5-year survival in patients treated with resection for carcinoma of the oesophagus and gastric cardia. INTRODUCTION: Surgery due to tumours of the oesophagus and gastric cardia is probably associated with the highest postoperative morbidity and mortality of all elective surgical procedures. Concentration to high-volume centres has been suggested to improve the outcome. MATERIALS AND METHODS: Between 1987 and 1996, all patients with squamous cell carcinoma or adenocarcinoma of the oesophagus or gastric cardia were identified from the Swedish Cancer Registry and the Swedish Hospital Discharge Registry. The study population was assessed according to patients operated at hospitals with a low (L-V), intermediate (I-V) or high operation volume (H-V), defined as <5 resections/year, 5-15 resections/year and >15 resections/year, respectively. We analyzed hospital mortality and 5-year survival. RESULTS: During the study period, 1429 patients were treated with resection for carcinoma of the oesophagus (n=665) or the gastric cardia (n = 764). A total of 74 hospitals were registered with at least one surgical resection, of which 90% performed <5 resections/year. The distribution of gender and age was comparable in the three groups. Hospital mortality was 10.4, 6.3 and 3.5% in the L-V, I-V and H-V groups, respectively. Overall 5-year survival was 17% (L-V), 19% (I-V) and 22% (H-V). Multivariate analysis showed an improved long-term survival for patients operated at H-V compared to L-V hospitals (p=0.02). CONCLUSION: This study supports an inverse relationship between hospital volume and hospital mortality after surgical tumour resection of the oesophagus or gastric cardia. Overall 5-year survival was significantly higher at high-volume hospitals compared to low-volume centres. We believe that concentrating these patients in high-volume hospitals is necessary to achieve high quality surgical treatment and to facilitate research aiming to improve prognosis. PMID- 15816606 TI - High exposure to polycyclic aromatic hydrocarbons may contribute to high risk of esophageal cancer in northeastern Iran. AB - BACKGROUND: The northeastern region of Iran has some of the highest rates of esophageal squamous cell carcinoma (ESCC) in the world. MATERIALS AND METHODS: To investigate the role of polycyclic aromatic hydrocarbons (PAHs) in the etiology of ESCC in northeastern Iran, we measured urine 1-hydroxypyrene glucuronide (1 OHPG), a stable PAH metabolite, in 99 inhabitants of this area. RESULTS: The median urine 1-OHPG in participants of this study was 4.2 pmol/ml. Forty-two subjects (42%) had levels ranging from 1 to 5 pmol/ml, indicative of moderate PAH exposure, and 41 (41%) had levels above 5 pmol/ml, indicative of very high exposure. Further analysis showed that 1-OHPG levels were high in all subgroups of our study subjects, including both sexes, rural and urban dwellers, and smokers and non-smokers. Only 15% of the variance in 1-OHPG was explained by age, sex, residence, smoking, nass, or opium consumption. This pattern of PAH exposure parallels the ESCC incidence pattern seen in this area. CONCLUSION: We conclude that people in northeastern Iran are exposed to widespread and very high levels of PAH, largely from unknown sources, and this may contribute to the high rates of ESCC observed in this area. PMID- 15816607 TI - Development of malignant mesothelioma during treatment for prolymphocytic leukemia: is asbestos or simian virus 40 a link? AB - A patient with a history of heavy asbestos exposure presented with B prolymphocytic leukemia/lymphoma (B-PLL). Soon after, he developed rapidly progressing malignant peritoneal mesothelioma. The concurrent development of both relatively uncommon diseases raised the possibility that a common causative factor might exist. Since asbestos, simian virus 40 (SV40), or both have been associated with lymphoproliferative disease and mesothelioma, we investigated both possible links in our patient. Imaging studies provided evidence for asbestos exposure because bilateral pleural plaques were identified. Tissues from bone marrow (involved with B-PLL) and from a peritoneal nodule (involved with mesothelioma) were examined for SV40 DNA using polymerase chain reaction (PCR): no SV40 DNA was detected. We conclude that asbestos remains the sole possible connection to both malignancies in this patient. It seems possible that fludarabine, an immunosuppressive chemotherapy, accelerated the occurrence and progression of malignant mesothelioma during the therapy for his B-PLL. PMID- 15816608 TI - Analysis of the response and toxicity to gefitinib of non-small cell lung cancer. AB - BACKGROUND: Gefitinib is an oral agent that inhibits the tyrosine kinase of epidermal growth factor receptor (EGFR), which had antitumor activity in patients with previously treated non-small cell lung cancer (NSCLC). We analyzed the efficacy, toxicity and overall survival time of gefitinib treatment in patients with NSCLC. PATIENTS AND METHODS: One hundred and twenty-two patients with NSCLC, who received gefitinib between 2002 and 2004 in our institutes, were evaluated retrospectively. RESULTS: The objective response rate was 24.6%. The variables identified as significant in univariate analysis included gender and smoking habit. The median overall survival time was 14.4 months. Significant variables associated with improved survival included good performance status (PS), female, adenocarcinoma and never smoked status, while never smoked status and good PS were independent prognostic factors in multivariate analysis. Four patients (3.3%) developed interstitial pneumonitis associated with gefitinib. CONCLUSION: Gefitinib showed favorable anti-tumor activity in females, never smokers and adenocarcinoma. PMID- 15816609 TI - Effects of PSK on T and dendritic cells differentiation in gastric or colorectal cancer patients. AB - BACKGROUND: Vaccine therapy targeting tumor antigens recognized by cytotoxic T cells (CTL) has been tried extensively. However, in a cancer-bearing state, the Th1/Th2 balance shifts to Th2 dominance, and this has been the obstacle to vaccine therapy to induce the CTL. DC1/DC2 subsets have also been reported to control the differentiation of Th subsets. The key to tumor immunotherapy is how to activate the DC1-Th1 lineage. PATIENTS AND METHODS: Six normal adults and 14 patients with gastric or colorectal cancers, who gave informed consent, were studied. The Th1/Th2 and DC1/DC2 ratios were determined by FACS. IL-12 and IL-10 production from PBMC were measured by ELISA. RESULTS: The Th1/Th2 and DC1/DC2 ratios were all significantly lower in the patients with gastric or colorectal cancers compared to normal adults. After protein-bound polysaccharide K (PSK) therapy in cancer patients, the Th1/Th2 balance shifted to Th1 dominance and the DC1/DC2 balance to DC1 dominance. IL-10 production was significantly decreased by PSK therapy. CONCLUSION: In the cancer-bearing state, the Th1/Th2 and DC/1/DC2 balance becomes Th2- and DC2-dominant. PSK therapy results in a shift of the Th1/Th2 and DC1/DC2 balance towards Th1 and DC1 dominance. We plan to examine whether combining dendritic cells (DC) vaccination therapy with oral PSK enhances the induction of T cell and DC differentiation in cancer patients. PMID- 15816610 TI - Circulating hormone levels in breast cancer patients. correlation with serum tumor markers and the clinical and biological features of the tumors. AB - BACKGROUND: Breast cancer grows in a hormone-rich environment which influences its biological features and thus, ultimately, its clinical behavior. Circulating hormone levels were measured in premenopausal and postmenopausal breast cancer patients prior to surgery, and correlated with all available clinical and biological features of the tumors and with serum tumor marker levels. MATERIALS AND METHODS: FSH, LH, 17-beta-estradiol, progesterone and prolactin were measured in 112 previously untreated breast cancer patients (54 premenopausal and 58 postmenopausal). Serum tumor markers (CEA, CA 15.3, CA-125 and Ca19.9) were measured at the same time. All tumors were studied after surgery for hormone receptor (ER and PR), Ki67, c-erb-B2 and p53 expression by means of immunohistochemistry, and for DNA-ploidy by means of flow cytometry. RESULTS: In premenopausal patients, high gonadotropin levels correlated directly with c-erb B2 overexpression by the tumors (FSH: p=0.02; LH: p=0.05). High estradiol levels correlated inversely (p=0.009) with Ki67 expression. In postmenopausal patients, high estradiol levels were inversely related to c-erb-B2 expression by the tumors (p=0.03), and high progesterone levels were also inversely related to Ki67 expression by the tumors (p=0.05). FSH levels correlated inversely (p = 0.02) with circulating carcinoembryonic antigen (CEA) levels. CONCLUSION: Circulating estradiol levels seem to be associated with a less proliferative breast cancer phenotype. FSH and LH levels, on the other hand, seem to exert dual actions in premenopausal and postmenopausal breast cancer patients. PMID- 15816611 TI - Adenoid cystic carcinoma of the breast: truly uncommon or easily overlooked? AB - BACKGROUND: Adenoid cystic carcinoma of the breast is an uncommon histologic form of breast cancer, comprising in most series less than 1% of all mammary cancers. Due to the rarity, little information about its presentation on image studies has been noted in the literature. Here we report two additional cases with emphasis on the intriguing image presentations. CASE ONE: A 67-year-old woman came to our clinic with the chief complaint of mastodynia. No obvious palpable mass of breast was found on physical examination. Mammography showed a small well-defined nodule in the medial part of the left breast without mammographic evidence of malignancy. Ultrasonography showed a 1.5 cm nodule with well-defined margin and heterogenous echogenicity in the medial part of the left breast. Unusually, a painful sensation was experienced on compression by the probe. The final pathological report was adenoid cystic carcinoma. CASE TWO: A 48-year-old woman also came to our clinic with the chief complaint of mastodynia. No obvious palpable mass of breast was found on physical examination. Mammography showed dense mammary tissue with no mammographic evidence of malignancy. Ultrasonography showed two contiguous well-defined nodules with heterogenous echogenicity in the upper, middle part of the left breast. Unusually, a painful sensation was also noted on compression by the probe. Histopathological examination showed typical features of an adenoid cystic carcinoma. CONCLUSION: Adenoid cystic carcinoma of the breast fails to show the typical appearance of invasive ductal carcinoma on both mammogram and ultrasonography, probably due to its relatively well-defined nature with less surrounding architectural disruption and fibrosis. Hence a "negative" finding or a benign-looking breast lesion on mammography cannot completely exclude the existence of this disease. The presence of a painful breast lesion without obvious inflammatory evidence while compressed is a meaningful clue, which should lead to the suspicion of adenoid cystic carcinoma of the breast. PMID- 15816612 TI - Occult cancer detected by positron emission tomography/computed tomography image fusion. AB - A 60-year-old male in a high-risk group for lung cancer, who also had positive sputum cytology, underwent examination by chest plain radiography, chest computed tomography (CT) and bronchofiberscopy. However, initially no abnormal findings were detected with these diagnostic modalities. Furthermore, thin-section spiral chest CT disclosed only a slight thickness of the right side of the tracheal wall, while consequent re-examination by bronchofiberscopy revealed only a light ulcerative lesion in the same area. However, the existence of a primary tumor was not confirmed using these modalities. Therefore, FDG-PET (positron emission tomography with fluorodeoxyglucose) was performed and demonstrated accumulation corresponding to the anterior mediastinum, although the exact location of the tumor was unclear by FDG-PET alone. Therefore, FDG-PET/CT image fusion was performed and resulted in the detection of a tracheal carcinoma on the outside of the right side of the tracheal wall. The patient then underwent tracheal sleeve resection including the tumor and tracheoplasty accompanied with wrapping using the flap of the thymus right lobe. To the best of our knowledge, this is the first reported case of tracheal carcinoma detected by FDG-PET/CT image fusion with consequent radical resection. PMID- 15816613 TI - Role of tumor-associated macrophages (TAM) in advanced gastric carcinoma: the impact on FasL-mediated counterattack. AB - BACKGROUND: Exactly what role does tumor-derived Fas ligand (FasL) play in cancer: maintaining the immune privilege site or inducing a pro-inflammatory effect? One possible hypothesis is that tumor-associated macrophages (TAM) act as the mediator that enables apoptosis of anti-tumor immune cells without FasL related inflammation. We have evaluated the tumor FasL expression and TAM along the tumor margin and/or in cancer stroma, and their impact on the infiltration of immune-competent cells into the tumor nest. PATIENTS AND METHODS: Tissue specimens from consecutive 84 advanced gastric carcinoma patients, who had undergone a curative resection, were evaluated for TAM (CD68+ cells), tumor FasL expression and immune status (CD8 + T cells). RESULTS: A high number of TAM significantly correlated with lymph node metastasis, intestinal type tumor and FasL expression. Although TAM had a tendency for an inverse correlation with the number of CD8+ T cells within the tumor nest (nest CD8) (p=0.0592), there was no correlation between FasL expression and nest CD8 (p=0.2158). This inverse association was found to be stronger in cases with both FasL-positive and high TAM tumors than in others (p=0.0139). The combination parameter of FasL-positive and high TAM became an independent prognostic factor in Cox's multivariate analysis, along with the pT status, nest CD8 and tumor cell apoptosis. CONCLUSION: We suggest that TAM works harmoniously with tumor-derived FasL and serves as a barrier against the infiltration of CD8+ T cells into the cancer nest. PMID- 15816614 TI - Efficacy of gemcitabine--cisplatin treatment in stage IIIA ("bulky"N2), IIIB and IV non-small cell lung cancer. AB - The objective of the study was to assess the efficacy of a gemcitabine and cisplatin combination for patients with stage IIIA"bulky"N2, IIIB or IV non-small cell lung cancer(NSCLC). PATIENTS AND METHODS: Patients with histological and/or cytological diagnosis of NSCLC were administered gemcitabine 1250 mg/m2 on days 1 and 8, and cisplatin 70 mg/m2 on day 1, every 3 weeks. RESULTS: One hundred and twenty patients with NSCLC, with median age of 53 years, and a WHO performance status of 0 (26%) or 1 (74%), were evaluated. The overall response rate was 40.0% with 37.5% partial response (PR) and 2.5% complete response (CR). Also, 38% of the patients had either minimal response (MR) or stable disease (SD). The median survival was 54.9 weeks. The time to progression was 28.1 weeks. There was no treatment-related death in this series. CTC grade 3/4 neutropenia occurred in 4.4% of the patients, while febrile neutropenia developed in 0,9% of the patients. CTC grade 4 thrombocytopenia occurred in 2.2%, and CTC grade 3/4 anemia developed in 3.3%. CONCLUSION: Our results support that gemcitabine and cisplatin administered as a 3-week cycle is an effective and safe regimen for the treatment of locally advanced or metastatic NSCLC. PMID- 15816615 TI - Continuous hepatic arterial infusion chemotherapy for liver metastasis from biliary tract and pancreatic cancers. AB - BACKGROUND: The efficacy of intrahepatic arterial chemotherapy for liver metastasis from biliary tract or pancreatic cancer remains uncertain. PATIENTS AND METHODS: Five patients with bilio-pancreatic liver metastasis underwent continuous hepatic arterial infusion chemotherapy. One treatment course basically consisted of a 14-day infusion period during which continuous infusions of 5 fluorouracil and intermittent bolus injections of cisplatin were given, and a subsequent 14-day intermission. After two consecutive courses, these drugs were administered bi-weekly. RESULTS: One complete and three partial responses were observed (response rate, 80%). In responders, the responses persisted until or even after the cessation of chemotherapy. The median survival was 15 months after the start of chemotherapy. The longest survivor has been disease-free for 46 months since a liver tumour remaining despite chemotherapy was eradicated by further treatment. Toxicity was acceptable. CONCLUSION: 5-Fluorouracil and cisplatin-based continuous hepatic arterial infusion chemotherapy may serve as a promising treatment for bilio-pancreatic liver metastasis. PMID- 15816616 TI - A computer-aided diagnosis (CAD) system in lung cancer screening with computed tomography. AB - We evaluated a computer-aided diagnosis (CAD) system with automatic detection of pulmonary nodules for lung cancer screening with computed tomography (CT). Five hundred and eighteen participants were examined with low-dose helical CT during a lung cancer screening by three respiratory physicians according to the General Rule edited by the Japan Lung Cancer Society. Four cases were detected by CAD and pathologically diagnosed as lung cancer. We compared the detection capability of the physician and CAD in 301 participants. Three physicians determined 75/301 (24.9%) participants as "e" (suspicious of lung cancer) in consensus without CAD, while 3 participants were added to "e" with CAD. Three physicians did not independently judge as "e" in 14 (18.7%), 16 (21.3%) and 16 (21.3%) out of 75 participants. CAD could not identify 17 (22.7%) nodules of 75 participants, and all 17 were less than 6 mm in diameter. The CAD system offers a useful second opinion when physicians examine patients at lung cancer CT screenings. PMID- 15816617 TI - Serum tenascin-C as a potential predictive marker of angiogenesis in non-small cell lung cancer. AB - BACKGROUND: Tenascin (Tn)-C is an extracellular matrix protein that is involved in tissue interactions during fetal development and oncogenesis. However, the role of serum Tn-C in non-small cell lung cancer (NSCLC) has not been clarified. PATIENTS AND METHODS: In this study, we determined the serum levels of Tn-C among NSCLC patients who underwent surgery, as well as other factors implicated for angiogenesis, to address the clinical implications in NSCLC. RESULTS AND CONCLUSION: The median concentration of serum Tn-C in NSCLC patients was slightly higher than that of normal controls, but this difference was not statistically significant. There was a positive correlation between serum Tn-C levels and microvessel density (MVD), serum osteopontin (OPN) and vascular endothelial growth factor (VEGF). In contrast, there was no correlation between serum Tn-C levels and serum carcinoembryonic antigen (CEA) and sialyl lewis-X (SLX) levels. The overall survival of patients with low Tn-C levels (<96 ng/ml) was significantly greater than that of patients with high Tn-C levels (> or =96 ng/ml). Intratumoral Tn-C expression was co-localized with expression of microvessels in the stroma of the cancer cells by immunohistochemical analysis. Moreover, enhanced in vitro migration of human umbilical vascular endothelial cells (HUVEC) was induced by recombinant Tn-C. Collectively, Tn-C may play an important role in angiogenesis of patients with NSCLC, and the determination of serum Tn-C may be useful in predicting intratumoral vasculature and patients' prognosis. PMID- 15816618 TI - A risk model for non-small cell lung cancer using clinicopathological variables, angiogenesis and oncoprotein expression. AB - BACKGROUND: The aim of this study was to investigate new prognostic factors, by using a prognostic model, that could help to identify the patient group with the greatest probability of death. PATIENTS AND METHODS: First, the clinicopathological variables were analyzed. Second, microvessels were immunohistochemically (IHC) stained with factor VIII-related antibody and then counted in the most intense vascularization area or hotspot, using an automatic image analyzer. In addition, biological angiogenesis-related factors such as vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase expression (iNOS) were also studied. Finally, we evaluated the IHC expression of p53 and p21WAF1 tumor supressor proteins. RESULTS: The significant independent predictors were: tumor size (p=0.0063), angiogenesis (p=0.0271) and p21WAF1 (p=0.0478). Thus, the most important factor was tumor size 2.7462 [95% CI=1.3307 5.6673]. Finally, these variables were included in a risk model, in order to identify the group with the highest associated probability of death. CONCLUSION: The analysis of several prognostic factors could establish a more accurate patient risk profile. PMID- 15816619 TI - Malignant fibrous histiocytoma of the breast. Report of two cases and review of the literature. AB - Sarcomas of the breast are uncommon, accounting for less than 1% of all primary malignancies. Among these tumors, malignant fibrous histiocytoma (MFH) is very rare. Two cases of this neoplasm are reported, with histological findings and surgical treatment. The issue of radiation-induced lesions after surgery for carcinoma and the necessity for a correct preoperative diagnosis is examined. A review of the available literature evaluates the histopathological and biological features of MFH of the breast, for which there are no prospective trials, owing to the rarity of this kind of neoplasm. The extent of surgery or role of axillary lymph nodes dissection and multimodality therapy are discussed. PMID- 15816620 TI - Telomerase activity in thyroid neoplasms evaluated by the expression of human telomerase reverse transcriptase (hTERT). AB - BACKGROUND: The recent availability of a monoclonal antibody against human telomerase reverse transcriptase (hTERT) has enabled us to investigate the telomerase activity of tissue specimens more easily than by classical TRAP assay. In this study, we studied teromerase activity in thyroid tumors using this antibody. MATERIALS AND METHODS: We immunohistochemically investigated hTERT expression in 166 thyroid neoplasms. RESULTS: Normal follicular cells did not express hTERT. In papillary carcinoma, high hTERT expression was observed in 34.5% of cases and it was directly linked to stage and the presence of lymph node metastasis. In follicular carcinoma, 39.2% were classified as the high hTERT expression group, showing a significantly higher incidence than that in follicular adenoma, 9.8%. In anaplastic carcinoma, 73.7% were regarded as the high hTERT expression group, which was larger than the percentages in papillary and follicular carcinomas. We investigated the diagnostic usefulness of hTERT measurement for discriminating follicular carcinoma from adenoma. Its specificity and positive predictive value were high at 90.2% and 83.3%, respectively, while the sensitivity and negative predictive value were low. CONCLUSION: These results suggest that: i) telomerase activity contributes to anaplastic transformation of differentiated carcinoma and ii) hTERT measurement may contribute to diagnosing follicular carcinoma, but whether it can be applied to preoperative diagnosis by fine-needle aspiration biopsy specimens remains unclear. PMID- 15816621 TI - Epithelial splenic cysts. AB - BACKGROUND: Epithelial splenic cysts (ESC) are uncommon lesions of the spleen. The etiopathogenesis of these cysts is controversial, even if Burrig's theory is the most accredited. The histological distinction between epidermoid and mesothelial cysts may be difficult, particularly for monostratified epithelium. PATIENTS AND METHODS: In the period between January 1986 and February 2004, 11 patients with ESC were studied. The history, physical findings, all relevant diagnostic studies and treatment were reviewed. All histological material was reviewed in detail with immunohistochemistry for CEA, CA 19-9, cytokeratin and calretinin. RESULTS: Epidermoid cysts were positive for CEA, CA 19-9, and cytokeratin, but negative for calretinin. Mesothelial cysts were positive for cytokeratin and calretinin, but negative for CEA and CA 19-9. CONCLUSION: Immunohistochemistry allows differential diagnosis between epidermoid and mesothelial cysts. With regard to etiopathogenesis, these data could mean that epidermoid and mesothelial cysts have distinct origins, though at variance with Burrig's theory. Although the ESC in this series were treated by open splenectomy, the recent approach by conservative and laparoscopic techniques offers great promise. PMID- 15816622 TI - Acute myopathy in a patient with lung adenocarcinoma treated with gemcitabine and docetaxel. AB - An extremely rare case of acute inflammatory myopathy during combination chemotherapy with gemcitabine and docetaxel for advanced non-small cell lung cancer in a 57-year-old diabetic male patient is reported. Despite the early clinical partial response of the underlying malignancy to the chemotherapeutic regimen, the patient developed symmetrical, painful, proximal muscle weakness in the lower limbs with peripheral edema after the administration of the fourth cycle of treatment. The syndrome regressed definitely after the discontinuation of chemotherapy and the administration of corticosteroids. The diagnosis of drug induced myositis is supported after the exclusion of other possible diagnoses. PMID- 15816623 TI - Metastatic rhabdomyosarcoma to the breast. AB - Secondary malignancy metastatic to the breast is uncommon, with an incidence of 0.5% to 3% of patients with extramammary malignancy. Although rhabdomyosarcoma is a common aggressive primary malignancy in the pediatric age group, metastatic deposits to the breast rarely occur and are mainly seen in adolescent girls. Here, we report an intriguing, rare adult case with metastasis to the breast from nasal rhabdomyosarcoma. A 31-year-old woman with the complaint of right neck mass noted recently came to this hospital for help. She had a history of nasal malignancy treated with radiotherapy in another hospital three months previously. Physical examination revealed multiple neck masses at bilateral neck areas. Bilateral neck dissection was performed and rhabdomyosarcoma, metastatic to lymph node, was the final diagnosis. One year after operation, the patient felt a large lump in her left breast. Surgical excision was performed and histological analysis was consistent with rhabdomyoblastic origin. Secondary malignancy metastatic to the breast is uncommon, yet this entity does exist. In view of the therapeutic implication, a metastatic breast lesion should not be mistaken as the primary breast carcinoma. Only with the awareness of such a possibility can prompt diagnosis and optimal treatment be achieved. PMID- 15816624 TI - Stressful and adverse life experiences in patients with breast symptoms; a prospective case-control study in Kuopio, Finland. AB - BACKGROUND: Psychosocial stress is widely thought to play a role in the aetiology of cancer in general and breast cancer in particular. Many studies have investigated the association between stressful life events and risk of breast cancer. However, the field of psychosocial cancer research is often problematic and findings have been contradictory, varying from no association to strong association. This inconsistency in results may be explained by the fact that most of the epidemiological data available come from retrospective case-control studies. We have conducted this case-control study with a so called "limited prospective study design"to reduce the potential for recall bias. MATERIALS AND METHODS: This study is an extension of the Kuopio Breast Cancer Study. Women with breast symptoms were referred by physicians to the Kuopio University Hospital (Finland) and were asked to participate in this study. The women were interviewed and reports on adverse and stressful life events were obtained before any diagnostic procedures were done, so neither the investigator nor the subject knew the final diagnosis of breast symptoms at the time of the interview. The research method used was the semi-structured in-depth interview method. All study subjects were also asked to complete standardised questionnaires (Beck Depression Inventory, Spielberger Trait Inventory). RESULTS: The clinical examination and biopsy showed breast cancer (BC) in 34 patients, benign breast disease (BBD) in 53 patients, while 28 study subjects showed to be healthy (HSS). The results indicated that BC patients had had significantly (p=0.02) more very severe (Gr IV) and severe (Gr III) stress in the previous 10 years preceding the investigation than the BBD and HSS groups. The BC group also reported significantly more moderate or severe losses than the BBD or the HSS groups (p=0.0009). CONCLUSION: The results of this study support an overall association between stressful life events and breast cancer risk. The biological explanation of the overall association might be that stress disturbs various areas of the immune systems predisposing to neoplasia. PMID- 15816626 TI - Poor oral health as a risk factor for esophageal squamous dysplasia in northeastern Iran. AB - BACKGROUND: Northeastern Iran has one of the highest rates of esophageal squamous cell carcinoma (ESCC) in the world. The reasons for observing such high rates of ESCC in this area are not fully clear. The current study evaluates the association between some potential risk factors and squamous dysplasia, a precursor lesion of ESCC, in northeastern Iran. MATERIALS AND METHODS: Using logistic regression, we compared potential risk factors of ESCC in 124 individuals with dysplasia and 50 normal individuals from northeastern Iran. Case/control status was histologically proven in all 174 study participants. RESULTS: Poor oral health showed a dose-response association with ESCC risk (p for trend < 0.01). Edentulous individuals had 5-fold higher risk of dysplasia compared to participants with good oral health (p-value < 0.01). CONCLUSION: Our results are consistent with two other published studies that have shown oral health as a risk factor for ESCC. The high incidence of ESCC in northeastern Iran may be partly attributable to poor oral health. PMID- 15816625 TI - Polymorphisms in the DNA base excision repair genes APEX1 and XRCC1 and lung cancer risk in Xuan Wei, China. AB - The lung cancer mortality rate in Xuan Wei is among the highest in China and has been causally attributed to high exposure to indoor smoky coal emissions, which contain high levels of PAHs and can lead to modified bases. We studied genetic polymorphisms in four DNA base excision repair genes in a population-based case control study in Xuan Wei with 122 lung cancer cases and 122 controls. Homozygous carriers of the APEX1 148Glu variant had an increased risk (OR, 2.13; 95% CI, 0.96-4.74), whereas persons with the XRCC1 399Gln allele had a decreased risk (OR, 0.60; 95% CI, 0.35-1.02) of lung cancer compared with wild-type carriers. Subjects with both at-risk genotypes (APEX1 Glu148Glu and XRCC1 Arg399Arg) had a higher risk of lung cancer (OR: 3.34; 95% CI: 1.16-9.67). We found genetic variants in APEX1 and XRCC1 may alter the risk of lung cancer in a special population in China. PMID- 15816627 TI - Gefitinib ('Iressa', ZD1839) may restore chemosensitivity in NSCLC patients? AB - Gefitinib ('Iressa, ZD1839) has promising antitumor activity in non-small cell lung cancer (NSCLC). However, patients with advanced NSCLC have few treatment options available if they are refractory to gefitinib. We describe four cases of patients with advanced NSCLC who previously responded to gefitinib and obtained significant tumor regression through retreatment with other cytotoxic agents. Gefitinib might restore chemosensitivity to previously chemorefractory patients. PMID- 15816628 TI - Prostate cancer therapy: standard management, new options and experimental approaches. AB - The management of prostate cancer is one of the core tasks for urologists today. This review focuses on therapeutic options for the curative and palliative treatment of prostate cancer. Within the area of urological competence, radical prostatectomy remains the standard procedure for the curative treatment of localised prostate cancer. Recently, interest in minimally-invasive procedures such as brachytherapy, focused ultrasound and cryotherapy has increased considerably. Established palliative treatment strategies include hormonal treatment schemes as the gold standard. Chemotherapeutic regimes are used for the hormone refractory disease. In addition to both the standard and new urological therapeutic options, promising experimental systemic strategies for the generalised disease are presented in this review. PMID- 15816629 TI - FOLFOX versus FOLFIRI: a comparison of regimens in the treatment of colorectal cancer metastases. AB - Colorectal adenocarcinoma ranks second as a cause of death due to cancer in the Western world. Already at the time of the primary tumor, 15-25% of the patients present with liver metastases while another 20% will develop metastasis following treatment of the colorectal primary. Without any treatment the median survival after the detection of metastases is approximately 9 months, depending on the extent of the disease at the time of diagnosis. Clinical trials with the "FOLFOX and FOLFIRI families" of drugs, designed for the treatment of metastatic colorectal cancer, their results and the costs of each therapy are examined. For each drug, the cost/mg, the cost/mg/m2 and the cost/therapy (according to its duration) are evaluated according to the prices reported in the Italian Directory of Medicines and Manufacturers, 63rd Edition, November 2003. PMID- 15816630 TI - Gn-RH antagonist possible response, after Gn-RH agonist failure in a man with metastatic prostate cancer. AB - Gn-RH agonists or surgical castration are considered standard treatment for patients affected by metastatic prostate cancer. Despite greater cost, chemical castration is often considered the treatment of choice as it is psychologically better tolerated. We report our experience of one patient undergoing treatment with Gn-RH agonist who developed an early resistance to the administered drug, with serum testosterone levels within the range of normality. PMID- 15816631 TI - Synchronous collecting duct carcinoma and papillary renal cell carcinoma: a case report and review of the literature. AB - The coexistence of multiple and synchronous primary neoplasms in the same organ (including kidney) has only rarely been described in the literature. We herein present a case of collecting duct carcinoma (CDC) combined with papillary renal carcinoma (RCC) having a 57-month disease-free survival CDC is a rather rare and aggressive neoplasm of the kidney. Sharing probably the same embryological origin, synchronous or metachronous association with in situ orpapillary transitional cell carcinoma (TCC) may be found; association with RCC has been only once reported in the literature. The high incidence of c-erbB-2 oncogene amplification in CDC further characterizes this tumor as a separate entity from renal cell carcinoma, and shows some genetic characteristics in common with TCC. The histological diagnosis of Bellini CDC can be confirmed by the positive immunohistochemical staining with a collecting duct marker and distal tubule marker and negative staining with a proximal tubule marker. PMID- 15816632 TI - Immunohistochemical staining of cathepsins B, L and stefin A in human hypophysis and pituitary adenomas. AB - BACKGROUND: New biological markers are needed for diagnosis of atypical pituitary adenoma. The study aimed to evaluate cathepsins B and L and inhibitor stefin A in relation to their aggressive progression. PATIENTS AND METHODS: We evaluated 19 adenomas and 10 normal hypophyses. Adenomas were divided according to their histological features and according to their functional activity, e.g. hormones secretion. Immunohistochemical labelling was scored for cathepsins and stefin A. RESULTS: High immunohistochemical scores for cathepsins B and L were more frequent in atypical pituitary adenoma. Higher cathepsin B scores were also observed in functional compared with non-functional tumours, independently of their histology. Stefin A labelling was observed in 90% of normal hypophyses, but only in 10% of adenomas: CONCLUSION: The levels of cathepsins B and L antigens are significantly higher in histologically atypical adenomas. Cathepsin B is also a marker of functional activity of the neoplastic glands. Therefore, we propose that this enzyme is evaluated as a diagnostic marker for tumour progression in non-functional adenomas, to distinguish atypical from benign tumours. PMID- 15816633 TI - Color-doppler using contrast medium in evaluating the response to neoadjuvant treatment in patients with locally advanced breast carcinoma. AB - BACKGROUND: The aim of our study was to assess the color-Doppler ultrasound (CDU) pattern in the analysis of neoadjuvant preoperative treatment of patients with locally advanced breast carcinoma, improvement after injection of contrast medium (Levovist) and possible correlations between morphological and vascular aspects of the neoplasm and postoperative histopathological findings. MATERIALS AND METHODS: We studied 50 patients affected by locally advanced breast carcinoma (T3a e b-T4), using CDU before and after injection of Levovist, prior to and after neoadjuvant chemotherapeutic treatment. RESULTS: The use of Levovist for ultrasound examinations prior to treatment revealed a higher number of vascular signals in 94% of the lesions compared to the basic color-Doppler examination; in only 3 cases (6%) were no modifications observed after injection of the contrast medium. This finding was also evident after neoadjuvant treatment, as a greater number of vessels in 28 lesions were observed, in addition to residual vascularization in 9 patients in whom the basic color-Doppler examination demonstrated substantial avascularity. Histopathology revealed that this method was more sensitive in disclosing the presence of active neoplastic tissue. CONCLUSION: Color-Doppler ultrasound is the first step in assessing the efficacy of neochemotherapeutic treatment in patients affected by locally advanced breast carcinoma. Levovist increases sensitivity and improves the diagnostic precision, thus allowing for a better image of the vessels, which is an important index of the biological activity of the neoplasm, compared to the basic color-Doppler examination. PMID- 15816634 TI - Influence of complementary Viscum album (Iscador) administration on microcirculation and immune system of ear, nose and throat carcinoma patients treated with radiation and chemotherapy. AB - With the techniques of vital microscopic and reflection spectrometric imaging, representative characteristics of microcirculation and immunology of white blood cells were evaluated before, during and after radiotherapy and chemotherapy of patients suffering from ear, nose and throat carcinomas. Adverse effects of radiotherapy and chemotherapy on the microcirculation and the immune system were decreased and reconstitution processes were accelerated by complementary administration of a standardized mistletoe extract (Iscador). PMID- 15816635 TI - Correlation between serum estradiol/progesterone ratio and survival length in invasive squamous cell cervical cancer. AB - BACKGROUND: There is epidemiological and laboratory evidence for an association between invasive squamous cell cervical cancer and female sex steroid hormones, such as oral contraceptives. MATERIALS AND METHODS: Premenopausal (n = 72) and postmenopausal (n=118) women with invasive squamous epithelial cervical cancer were included in this study. Serum estradiol and progesterone and DNA S-phase fraction as a measure of proliferative activity were analysed, in 51 pre-, and 77 postmenopausal women, among whom 13 and 43, respectively, died from disease. RESULTS: There was a significant positive correlation between a low serum estradiol/progesterone ratio and short survival in those premenopausal women who eventually died from cancer (p=0.02). Clinical stage was similar when the estradiol/progesterone ratio was dichotomized. There was no association between estradiol/progesterone ratio and survival-months in postmenopausal women. In both pre- and postmenopausal women deceased from cervical cancer, a S-phase fraction at or above 12% was correlated with reduced survival-months (p=0.03). CONCLUSION: These results, if confirmed, contribute to bridging the gap between previous epidemiological and laboratory findings of an association between female sex steroid hormones and squamous cell cervical cancer. PMID- 15816636 TI - CD44v6 as a prognostic factor in cervical carcinoma FIGO stage IB. AB - The aim of our study was to clarify whether CD44v6 evaluation can serve as a universally applicable prognostic factor in patients with FIGO stage IB cervical carcinoma. A retrospective study was performed on 178 FIGO stage IB (142 IB N0, 36 IB N1) radically operated cervical carcinoma patients. The expression of CD44v6 was investigated by immunohistochemistry (IHC). The prognostic significance of established prognostic factors and CD44v6 expression was analyzed by univariate and multivariate analyses. To test the reproducibility and to account for interobserver variability, all specimens were evaluated independently at two institutions. Two different IHC scoring systems, several cut-off levels for CD44v6 positivity and several statistical methods for IHC results evaluation were used. In a univariate analysis, the most significant prognostic factor for overall survival (OS) was lymph node status (p<0.001) followed by tumor volume, LVSI, GOG score (p<0.01) and a deep stromal invasion (p = 0.06). We found a strong correlation between CD44v6 expression and squamous cell carcinoma (SCC) (SCC vs. adenocarcinoma - p<0.001) and between CD44v6 expression and deep stromal invasion, LVSI and GOG score (p<0.05). The CD44v6 expression was not a statistically significant prognostic factor for OS in a univariate analysis (p=0.39 Vienna; p=0.54 Freiburg). In a multivariate analysis, the most significant prognostic factor for OS was lymph node status (p =0.002), followed by tumor diameter and LVSI (p<0.05). CD44v6 expression was not a statistically significant prognostic factor for OS or disease-free interval (DFI) independent of the scoring method used. In conclusion, we demonstrated that CD44v6 expression is associated with LVSI, deep stromal invasion and SCC, but has no prognostic influence on OS and DFI in a population of 178 women with FIGO stage IB cervical carcinoma. PMID- 15816637 TI - Tissue inhibitor of metalloproteinases-4 is expressed in cervical cancer patients. AB - BACKGROUND: Tissue Inhibitors of Metalloproteinases (TIMPs) play a critical role in extracellular matrix remodeling, which is involved in tumor growth and metastasis. Elevated TIMP levels are reported in association with cancer progression. In particular, it has been shown that TIMP-1 and -2 levels are increased in cervical cancer patients. We analyzed, for the first time, TIMP-4 expression in cervical tumor samples. MATERIALS AND METHODS: Semiquantitative RT PCR was performed in 26 tumor and 6 normal cervical samples. RESULTS: The study included 32 samples, 7 IB samples, 9 IIB samples, 10 IIIB samples and a control group (n =6) of normal cervical squamous epithelial tissues. Whereas none of the control samples expressed TIMP-4, 24 (88%) of the 26 cervical cancer samples expressed the inhibitor. Higher TIMP-4 levels were found in advanced stage disease (p=0.016, Chi-square test). CONCLUSION: TIMP-4 is expressed de novo in cervical cancer. Higher inhibitor expression levels were found in stages II and III. PMID- 15816638 TI - Addition of oxaliplatin to neo-adjuvant radiochemotherapy for irresectable rectal cancer, a phase I study. AB - BACKGROUND: The aim was to determine the maximum tolerable dose of oxaliplatin added to (oral) 5FU in irresectable rectal cancer. PATIENTS AND METHODS: Nineteen patients were treated; 13 patients received 5FU/LV and 6 patients capecitabine. Oxaliplatin was administered on days 1 and 29 at dose levels 85 and 130 mg/m2. Four to seven weeks thereafter, surgery was performed. RESULTS: In 6 patients treated with 85 mg/m2, one grade 3 elevation of liver transaminases occurred. Of 7 patients who received 130 mg/m2, 1 patient experienced a grade III thrombocytopenia and 1 patient died of neutropenic fever, probably due to an urosepsis. Six patients were treated with capecitabine, of whom 3 developed a grade III gastrointestinal toxicity. An R0 resection could be performed in 93%, a pT0-2N0 in 39%, with 2 pCR's. CONCLUSION: The addition of oxaliplatin at 85 mg/m2 on days 1 and 29 to radiotherapy and 5FU/LV or capecitabine in irresectable rectal cancer is feasible. PMID- 15816639 TI - Four tumour markers for urinary bladder cancer--tissue polypeptide antigen (TPA), HER-2/neu (ERB B2), urokinase-type plasminogen activator receptor (uPAR) and TP53 mutation. AB - PURPOSE: Tissue polypeptide antigen (TPA) is present in the proteolytic fragments of cytokeratins 8, 18 and 19 as a component of the cytoskeleton of nonsquamous epithelia. HER-2/neu protein is a transmembrane tyrosine kinase cell surface growth factor receptor that is expressed on normal epithelial and some cancer cells. The urokinase-type plasminogen activator receptor (uPAR) is a GPI-linked single-chain glycoprotein. Mutations of the tumour suppressor gene P53 (TP53) are frequently correlated with tumour development and progression. We compared TPA, HER-2/neu and uPAR, and TP53 mutation in tumour-free and bladder cancer patients. MATERIALS AND METHODS: Clinical samples were used from 60 patients with tumours of the urinary bladder and from 9 patients with benign urological diseases. TPA was analyzed by the immunoluminometric assay LIA-mat TPA-MProlifigen. HER-2/neu was measured using the Bayer Oncoprotein test. uPAR was measured with the IMUBIND Total uPAR ELISA Kit. Mutation status in TP53 exons 5, 6, 7 and 8 was analyzed by temperature gradient gel electrophoresis of exon-specific PCR products and by sequence analysis. Statistical analysis included ROC, Mann-Whitney U-test and Pearson's correlation. RESULTS: Pathological concentrations of TPA, HER-2/neu and uPAR are detectable in the serum and in urine of bladder cancer patients. The calculated diagnostic sensitivity for TPA in serum was 68.37%, for TPA in urine 33.3%, for HER-2/neu 86.7% and for uPAR 79.5%. Pathological levels of TPA in serum (p=0.001) and HER-2/neu (p =0.001) were significantly higher in patients with bladder cancer in comparison to the control group. For superficial bladder cancer, the mutation frequency in TP53 was 50%, while for invasive bladder cancer the mutation frequency in TP53 was 100%. Elevated TPA, HER-2/neu and uPAR levels were associated with all grades and stages of bladder cancer. CONCLUSION: TPA, HER-2/neu or uPAR can differ between bladder cancer patients and the control group, but not between superficial and invasive bladder cancer. TP53 mutation frequently occurs in higher stages of bladder tumours. PMID- 15816640 TI - The spleen plays an immunosuppressive role in patients with gastric cancer: involvement of CD62L+ cells and TGF-beta. AB - CD62L is the human homologue of the murine lymphocyte homing receptor, mel-14. We investigated CD62L + cells in the spleen from patients with gastric cancer. Flow cytometric analysis revealed that CD62L + cells were decreased in the peripheral blood, but inversely increased in the spleen in parallel with disease progression in gastric cancer patients. The increased CD62L+ cells resided in the CD4+ suppressor-inducer phenotype, and the removal of CD62L+ cells from spleen cells resulted in a decrease of concanavalin-A-induced suppressor activity in vitro in one-way allogeneic mixed lymphocyte reaction. The CD62L+ cells included CD4+CD25+ regulatory T cells. The culture supernatant of CD62L + cells showed TGF-beta activity that permitted anchorage-independent growth of normal rat kidney (NRK) cells in a soft agar. TGF-beta activity was more significantly detectable in the splenic vein than in the peripheral blood, and TGF-beta mRNA was detectable in the spleen from advanced gastric cancer patients. These results suggest that CD62L+ cells migrate into the spleen with disease progression of gastric cancer and serve as suppressor-inducer cells with TGF-beta production to induce regulatory T cells, contributing to disease-associated immunosuppression in advanced gastric cancer patients. PMID- 15816641 TI - Pneumocystis carinii pneumonia in patients with solid tumors and lymphomas: predisposing factors and outcome. AB - AIM: The incidence of Pneumonocystis carinii pneumonia (PCP) is increasing in patients with cancer. The possible routes of transmission in this population, as well as the epidemiological data of PCP, are not very well understood. The collection and analysis of data concerning the predisposing factors for PCP will elucidate this subject. PATIENTS AND METHODS: We studied 26 patients suffering from cancer who developed PCP during the five-year period 1997--2002. RESULTS: Twenty-one patients had a solid tumor diagnosis while the remaining five had a lymphoma. All of them received intensive combination chemotherapy, while eight of them also received high-dose therapy and bone marrow transplantation. Nineteen of our patients had long hospitalization before the onset of PCP. All of them had received corticosteroids for various reasons. Among them, many patients had been exposed to radiotherapy and, in particular, in fields which encompass the major thoracic duct. Eighteen patients survived after the initiation of appropriate treatment, while eight others succumbed. CONCLUSION: In this series, protracted deep lymphopenia, long hospitalization, radiotherapy and intensive chemotherapy were considered serious risk factors for developing PCP. PMID- 15816642 TI - Persistence of K-ras mutations in plasma after colorectal tumor resection. AB - Free DNA in the circulation is increased five-to ten-fold in patients with solid tumours compared to healthy controls. A range of tumor-specific mutated DNA has been shown to be readily extractable and possible to analyse from plasma and serum in these patients. K-ras oncogene mutations are an early event in a subset of colorectal tumors and have been found in 30-60% of patients with colorectal carcinoma (CRC). The presence of tumor-derived k-ras gene mutations in the circulation has previously been described before surgery. The aim of this study was to characterize the presence of mutant k-ras in plasma in the short-term postoperative period after radical surgery of CRC patients, and further to characterize this in relation to relapse of the disease. Tumors and corresponding plasma pre- and postoperatively on day three after surgery were collected from 25 patients with CRC (Dukes' stage A-D). Biopsies for DNA extraction from the tumors were collected from the most invasive parts microscopically. After PCR amplification of the k-ras gene (codon 12 and 13), the presence of mutations was analysed by TGGE (temperature gradient gel electrophoresis). Twenty four/25 patients underwent putatively curative resections. Sixteen of the 25 patients (64%) expressed k-ras mutations in their tumor. Of these, 9 patients (56%) also had detectable k-ras mutations in preoperative plasma samples. On day three postoperatively, 8 of these patients persistently were found to have mutant k-ras in the plasma. This was not correlated with tumor stage. None of the 9 tumor mutation-negative cases expressed mutated k-ras in their plasma pre- or postoperatively. The results indicate that plasma mutant k-ras can be detected pre- and early postoperatively in all stages of colorectal neoplasia. No correlation between short-term postoperative persistence of mutant plasma-DNA and disease recurrence at follow-up was found. However, the use of k-ras as a marker during postoperative follow-up and as a possible tool for early detection of recurrent disease must be further characterized. PMID- 15816643 TI - High-dose alkylating agents with autologous hematopoietic stem cell support and trastuzumab in ERBB2 overexpressing metastatic breast cancer: a feasibility study. AB - BACKGROUND: ERBB2 overexpression predicts a worse outcome for patients receiving high-dose chemotherapy (HDC). Trastuzumab improves response rate and survival in ERBB2 overexpressing metastatic breast cancer patients (MBC). We investigated the feasibility of combining high-dose alkylating agents with autologous hematopoietic stem cell (AHSC) support and trastuzumab in ERBB2 overexpressing MBC. PATIENTS AND METHODS: Eleven consecutive patients with pre-treated ERBB2 overexpressing MBC were enrolled. HDC regimen consisted of a single course of cyclophosphamide 120 mg/kg + melphalan 140 mg/m2 (CyMEL, n =8), a single course of Thiotepa 600 mg/m2 (TTP, n = 1) or a sequential combination of Thiotepa 600 mg/m2 followed on day 21 by BCNU 600 mg/m2 (TTP-BCNU, n =2). Trastuzumab (4mg/kg) was started 24 h after AHSC infusion and then administered weekly (2 mg/kg). RESULTS: Median time to neutrophil and platelet recovery was 10 and 14.5 days, respectively. Three patients experienced febrile neutropenia and in 2 Herpes virus infections were documented. Five grade III/IV mucositis/oesophagitis were recorded. One patient experienced a reversible atrial arrhythmia on day 2 of trastuzumab, and another patients had a nonsymptomatic decrease in LVEF >10% on week 12 of trastuzumab. No toxic death was recorded. Median time to progression was 5 months (1 to 38 +). CONCLUSION: Combining alkylating agent-based HDC and trastuzumab appears to be feasible in ERBB2 overexpressing MBC and warrants further investigation in a larger cohort. PMID- 15816644 TI - A new dose-intense epoetin alfa regimen effective in anemic cancer patients receiving chemotherapy: an open-label, non randomized, pilot study. AB - BACKGROUND: Chronic anemia is a well-recognized complication of both cancer and cytotoxic treatments and is associated with symptoms (e.g., fatigue, dyspnea) that may induce or exacerbate functional deterioration. The use of recombinant human erythropoetin (rHuEPO epoetin alfa) clearly increased haemoglobin (Hb) levels, decreased transfusion needs and allowed recovery of quality of life in anemic cancer patients (pts) undergoing chemotherapy (CT). The purpose of this open-label, non randomized, pilot study was to assess the safety and efficacy of an intensive 19-day epoetin alfa treatment in anemic patients with solid tumors receiving chemotherapy. PATIENTS AND METHODS: TREATMENT: patients received a single induction s.c. dose of epoetin alfa 40,000 IU day 1 and twice a dose of 10,000 IU s.c. (8.00 a.m.- 8:00 p.m.) on days 3, 5, 8, 10, 12, 15, 17 and 19. The total dose of epoetin alfa per patient was 200, 000 IU. Iron supplementation: 125 mg i.v. days and 8. Soluble transferrin receptor (sTfR) levels were performed on days 1,8 and 15. This epoetin induction regimen was not followed by an epoetin maintenance therapy. PATIENTS: Twenty-nine anemic (Hb< or =11.5 g/dL) pts with non myeloid malignancies undergoing CT were included in the study. RESULTS: At baseline the mean Hb level was 9.41 g/dl. On day 8, the mean Hb level increased to 10.07 g/dl (p<0.0001), reaching 10.68 g/dl on day 15 (p<0.0001). On days 22 and 29, the mean Hb levels increased to 10.93 and 11.05 g/dl, (p=0.002 and 0.033, respectively). No patient received blood transfusions. The global mean increase of Hb level was 1.64 g/dl (basal to d 29). It was defined as a major response: an increase of Hb levels > 1.5 g/dl. A rate of 62% (18/29 patients) of major responses was observed on day 21. Moreover, 25/29 patients (86.2%) presented an increase of Hb levels > 1 g/dl after 21 days. On days 8 and 15, the mean sTfR levels had increased significantly ( p=0.021 and 0.001, respectively). The increase of mean sTfR level after 15 days correlated significantly with the increase of mean Hb level in the first two weeks of epoetin therapy (p=0.05). Epoetin alfa has been well tolerated so far in the study. CONCLUSION: The results of the present study suggest that an induction dose of 40,000 IU of epoetin alfa, followed by 8 maintenance doses of 20,000 IU each, may improve the standard response in terms of both time to response and Hb increase. Moreover, the Hb levels seemed to increase after epoetin therapy discontinuation (d22-29). PMID- 15816646 TI - For AMA, McClellan inserts some pills in sugar. PMID- 15816645 TI - Differential cyclooxygenase-2 enzyme expression in radiosensitive versus radioresistant glioblastoma multiforme cell lines. AB - BACKGROUND: Glioblastoma multiforme (GBM) is a high-grade primary brain tumor that is refractory to current forms of treatment. In cell studies, the growth rate of GBM cells correlates with the level of Cyclooxygenase-2 (COX-2) enzyme expression. COX-2 has been implicated in carcinogenesis of systemic cancers. Recently, COX-2 inhibition has been shown to increase the radiosensitivity of various tumors. We wished to assess whether the expression of COX-2 is greater in radioresistant versus radiosensitive forms of GBM. MATERIALS AND METHODS: The radiosensitive (A172) and radioresistant (T98G) Glioblastoma multiforme cell lines were assayed for COX-2 expression using standard immunofluorescence histochemistry. Fluorescence readings were recorded per field. Western blot analysis was performed on both A172 and T98G GBM cell lines. The radioresistant cells were exposed to incremental doses of radiation in the presence and absence of a COX-2-selective inhibitor. Radioresistant cells were then exposed to incremental doses of COX-2-selective inhibitor at a constant dose of radiation. RESULTS: The radioresistant cell line T98G had an approximate 1. 7-fold greater expression of COX-2 than did the radiosensitive cell line A172, as per immunofluorescence histochemistry. Western blot analysis confirmed this finding. Statistical analysis (Bonferroni/Dunn) showed the results to be significant (p<0.0001). The wells containing radioresistant cells exposed to incremental doses of radiation and COX-2 inhibitors appeared to have higher cell kill when compared to radiation alone. Furthermore, increasing the COX-2 inhibitor concentration yielded higher cell kill. CONCLUSION: The results presented here show that the radioresistant GBM cell line, T98G, has a greater expression of COX 2 than does the radiosensitive GBM cell line, A172. These results suggest that: (i) COX-2 expression may serve as a marker for assessing radioresistance in GBM, (ii) COX-2 inhibition may lower the required doses of postoperative radiation, (iii) COX-2 inhibitors may have a role in radiosensitizing otherwise radioresistant forms of GBM. PMID- 15816647 TI - Infant mortality rates. PMID- 15816648 TI - Humour a welcome addition. PMID- 15816649 TI - Why do we receive Alberta RN and Canadian Nurse magazines in two separate mail outs? PMID- 15816650 TI - Going back to school. PMID- 15816652 TI - Medicare's "un-sustainable" growth rate. PMID- 15816651 TI - Strong women fuel passion for nursing. PMID- 15816653 TI - Changing the paradigm for transfusing the anemic patient in an acute care environment. AB - Blood is a precious and vital resource in medicine. Its importance in maintaining human life is second only to the oxygen it transports. Nearly 300,000 blood product transfusions occurred in Kentucky last year. While donor attrition from the pool is inevitable, recruiting efforts have resulted in an overall stable donor population in the state over the last five years. Roughly 5% of community members who are eligible to donate do so each year. Despite our effort to constantly recruit new donors, there is evidence to suggest that clinicians are less than rigorous in their efforts to conserve and to be accountable for this precious human product. For example, the indications for blood transfusions are often not documented in up to a third of the cases, and arbitrary transfusion "triggers" are often the criteria used to justify transfusion rather than the patient's clinical condition. Recent development of evidence-based criteria for prevention and treatment of anemia combined with technological advances suggest that a major paradigm shift in the clinical use of red cells is warranted. PMID- 15816654 TI - The relationship between elevated body mass index and lethal ischemic heart disease: an eleven-year retrospective review of medical examiners' adult autopsies in Kentucky. AB - Both childhood and adult overweight (OW) and obesity (OB) currently loom unabated at epidemic proportions in the United States (US), to which Kentucky (KY) is a leading contributor. These conditions are significant risk factors for development of ischemic heart disease (IHD), the primary cause of natural death in the US. These mortality statistics (see below), combined with Kentucky's significantly prevalent OW/OB population, prompted this investigation of the presumptive correlation between body mass index (BMI) and IHD in the medical examiners' office (MEO) autopsy cohort. The goals of this study were threefold: (1) to identify all adults between 19 and 85 years old autopsied over an 11-year period (1991-2001) at KY MEO whose deaths were attributed to IHD; (2) to establish the BMI (kg/m2 ), a gauge of fat mass, at death; and (3) to determine whether there is a statistically significant relationship between elevated (or "unhealthy") BMI--categorized as either OW or OB--and an increased risk for the development of IHD-associated premature death in KY. This report demonstrates a significant correlation between death in adulthood attributed to IHD and unhealthy BMI in KY autopsy cases in the MEO. PMID- 15816655 TI - Increasing awareness of health care resources for the uninsured and underinsured:a pilot study. AB - Programs are available to provide assistance to the uninsured, but information about how to access those programs is not readily available to the average uninsured citizen. A pilot study involving the University of Kentucky Family and Consumer Science Agents and Homemaker groups in two rural counties significantly increased the number of people who accessed one of the programs, Health Kentucky. Findings from the study could be validated in other Kentucky counties and with other types of health-related messages. PMID- 15816656 TI - Living without immunizations--a new growing trend. PMID- 15816657 TI - Intimate partner violence: homicides followed by suicides in Kentucky. PMID- 15816658 TI - Colorectal cancer facts. PMID- 15816659 TI - Commercial conflict of interest and continuing education. PMID- 15816660 TI - Oral healthcare for patients with disabilities. PMID- 15816661 TI - Review of hemostatic agents used in dentistry. PMID- 15816662 TI - Radiation therapy and oral care. PMID- 15816663 TI - Dental implant placement in the maxillary anterior region: guidelines for aesthetic success. PMID- 15816664 TI - Simplified adhesive cementation: what, where, and how. PMID- 15816665 TI - No-preparation porcelain veneers--back to the future! PMID- 15816666 TI - Conservatism and simplicity: aesthetic restoration of the single maxillary central incisor. PMID- 15816667 TI - Use of natural teeth as pontic sections in a bonded, lingually retained prothesis. PMID- 15816668 TI - Form and function: a balancing act. PMID- 15816669 TI - Many problems, mini solutions. PMID- 15816670 TI - Considerations in treatment planning of the periodontal patient. AB - Through the use of periodontal, orthodontic, and restorative procedures, this patient was restored to function with a stable dentition and minimal pocket depths. The patient was followed for 10 years. All areas exhibited a healthy periodontium with stability of all areas of the dentition. PMID- 15816671 TI - Too tired, too stressed, too busy to enjoy dentistry? PMID- 15816672 TI - Ten ways to improve your computer and network experience. PMID- 15816673 TI - Pulp chamber morphology: basic research leads to clinical technique. PMID- 15816674 TI - A new role for autotrophic methanogens? PMID- 15816675 TI - Optimization of enhanced biological phosphorus removal after periods of low loading. AB - Enhanced biological phosphorus removal is a well-established technology for the treatment of municipal wastewater. However, increased effluent phosphorus concentrations have been reported after periods (days) of low organic loading. The purpose of this study was to evaluate different operating strategies to prevent discharge of effluent after such low-loading periods. Mechanisms leading to these operational problems have been related to the reduction of polyphosphate accumulating organisms (PAOs) and their storage compounds (polyhydroxy alkanoates [PHA]). Increased effluent phosphorus concentrations can be the result of an imbalance between influent loading and PAOs in the system and an imbalance between phosphorus release and uptake rates. The following operating conditions were tested in their ability to prevent a reduction of PHA and of overall biomass during low organic loading conditions: (a) unchanged operation, (b) reduced aeration time, (c) reduced sludge wastage, and (d) combination of reduced aeration time and reduced sludge wastage. Experiments were performed in a laboratory-scale anaerobic-aerobic sequencing batch reactor, using acetate as the carbon source. Without operational adjustments, phosphorus-release rates decreased during low-loading periods but recovered rapidly. Phosphorus-uptake rates also decreased, and the recovery typically required several days to increase to normal levels. The combination of reduced aeration time and reduced sludge wastage allowed the maintenance of constant levels of both PHA and overall biomass. A mathematical model was used to explain the influence of the tested operating conditions on PAO and PHA concentrations. While experimental results were in general agreement with model predictions, the kinetic expression for phosphorus uptake deviated significantly for the first 24 hours after low-loading conditions. Mechanisms leading to these deviations need to be further investigated. PMID- 15816676 TI - Effect of bubble-induced surface turbulence on gas-liquid mass transfer in diffused aeration systems. AB - Models that predict volatilization of organic compounds from wastewater treatment basins may underestimate emission rates if the surfaces are considered as quiescent. In reality, the water surface may be agitated by subsurface aeration, increasing mass transfer across the tank surface air-water interface. This study investigated the effect of turbulence, induced by diffused bubble aeration, on mass transfer at the water surface of a pilot aeration basin. The mass transfer of ammonia from an enclosed headspace over the basin to acidified water was measured when different diffuser types and airflow rates were applied. Oxygen transfer tests were conducted immediately following each ammonia-transfer test. Increasing airflow rates through fine- and coarse-bubble diffusers had a significant effect on the ammonia mass-transfer rate. Experimental mass-transfer parameters (K(L)a's) for surface volatilization derived with aeration present were up to 48% higher than the K(L)a values for quiescent conditions over the range of conditions tested. No effect of diffuser type on ammonia transfer could be determined. The study results infer an effect on oxygen transfer into the water at the surface and potential transfer of volatile organic compounds, if present, from the water. The results of the ammonia mass-transfer experiments suggest that adjustments to the existing mass transfer correlations for surface volatilization from aeration basins may be in order. Such adjustments will have the greatest effect on predictions for the less volatile compounds, under conditions of low airflow rates. PMID- 15816677 TI - Development of a gravity-independent wastewater bioprocessor for advanced life support in space. AB - Operation of aerobic biological reactors in space is controlled by a number of challenging constraints, mainly stemming from mass transfer limitations and phase separation. Immobilized-cell packed-bed bioreactors, specially designed to function in the absence of gravity, offer a viable solution for the treatment of gray water generated in space stations and spacecrafts. A novel gravity independent wastewater biological processor, capable of carbon oxidation and nitrification of high-strength aqueous waste streams, is presented. The system, consisting of a fully saturated pressurized packed bed and a membrane oxygenation module attached to an external recirculation loop, operated continuously for over one year. The system attained high carbon oxidation efficiencies often exceeding 90% and ammonia oxidation reaching approximately 60%. The oxygen supply module relies on hydrophobic, nonporous, oxygen selective membranes, in a shell and tube configuration, for transferring oxygen to the packed bed, while keeping the gaseous and liquid phases separated. This reactor configuration and operating mode render the system gravity-independent and suitable for space applications. PMID- 15816678 TI - Retention of chromium (VI) on a macroporous char following ChemChar gasification and successive leaching with water and acids. AB - A granular macroporous char, triple-reverse-burn (TRB) char, was loaded with 23.40 mg Cr/g TRB char from an aqueous solution, and the retained metal was leached by water, 0.66 M nitric acid, concentrated nitric acid, and concentrated hydrochloric acid before and after treatment by a reductive thermal gasification process (ChemChar process developed by ChemChar Research, Inc., Columbia, Missouri). The chromium leachate was analyzed by flame atomic adsorption. Reverse and forward-mode gasifications were performed on the metal-laden char. With the exception of a 10% mass loss of carbon, the reverse mode gasification process does not change the physical characteristics of the granular char, but does increase the retention of the chromium from 16.7 to 24.2%, depending on the leachant. The forward mode gasification process produces a vitrified (or glasslike) ash residue. There was an 11.6 to 13.1% increase in the retention of the chromium by the slag and ash when compared to the nongasified chromium-loaded TRB char. Chromium (VI) was effectively removed from solution by TRB char and found to be retained to a higher degree on the char after a reductive thermal treatment. PMID- 15816679 TI - Field demonstration of the combined effects of absorption and evapotranspiration on septic system drainfield capacity. AB - Drainfields for disposal of septic tank effluents are typically designed by considering the loss of water by either upward evapotranspiration into the atmosphere or lateral and downward absorption into the adjacent soil. While this approach is appropriate for evapotranspiration systems, absorption systems allow water loss by both mechanisms. It was proposed that, in areas where high evapotranspiration rates coincide with permeable soils, drainfield sizes could be substantially reduced by accounting for both mechanisms. A two-year field demonstration was conducted to determine appropriate design criteria for areas typical of the Texas High Plains. The study consisted of evaluating the long-term acceptance rates for three different drainfield configurations: evapotranspiration only, absorption only, and combined conditions. A second field demonstration repeated the experiments for additional observation of the combined evapotranspiration and absorption and achieved similar results as the first study. The field tests indicated that the current design loading criteria may be increased by at least a factor of two for the Texas High Plains region and other Texas areas with similar soil composition and evapotranspiration rates, while still retaining a factor of safety of two. PMID- 15816680 TI - Evaluation of membrane bioreactor process capabilities to meet stringent effluent nutrient discharge requirements. AB - A six-stage membrane bioreactor (MBR) pilot plant was operated to determine and demonstrate the capability of this process to produce a low-nutrient effluent, consistent with the nutrient reduction goals for the Chesapeake Bay. Biological nitrogen removal was accomplished using a multistage configuration with an initial anoxic zone (using the carbon in the influent wastewater), an aerobic zone (where nitrification occurred), a downstream anoxic zone (where methanol was added as a carbon source), and the aerated submerged membrane zone. The capability to reliably reduce effluent total nitrogen to less than 3 mg/L as nitrogen (N) was demonstrated. A combination of biological (using an initial anaerobic zone) and chemical (using alum) phosphorus removal was used to achieve effluent total phosphate concentrations reliably less than 0.1 mg/L as phosphorus (P) and as low as 0.03 mg/L as P. Alum addition also appeared to enhance the filtration characteristics of the MBR sludge and to reduce membrane fouling. Aeration of the submerged membranes results in thickened sludge with a high dissolved oxygen concentration (approaching saturation), which can be recycled to the main aeration zone rather than to an anoxic or anaerobic zone to optimize biological nutrient removal. Biological nutrient removal was characterized using the International Water Association Activated Sludge Model No. 2d. The stoichiometry of chemical phosphorus removal was also consistent with conventional theory and experience. The characteristics of the solids produced in the MBR were compared with those of a parallel full-scale conventional biological nitrogen removal process and were generally found to be similar. These results provide valuable insight to the design and operating characteristics of MBRs intended to produce effluents with very low nutrient concentrations. PMID- 15816681 TI - Biogeochemical analysis of hydrogen sulfide removal by a lava-rock packed biofilter. AB - Although lava-rock-based biofilters have demonstrated their efficiencies for hydrogen sulfide (H2S) removal found in odorous air emissions, the biogeochemical basis for this removal is unclear. In this study, samples of lava rock and rinse water from biofilters at Cedar Rapids Water Pollution Control Facilities (Iowa) were used to study the structure and chemical composition of lava rock and to identify the predominant microorganism(s) present in lava-rock-based biofilters. It was found that iron, in the form of Fe2+ and Fe3+, was present in lava rock. Although literature suggests that Acidithiobacillus thiooxidans are primarily responsible for gaseous H2S removal in biofilters, our study showed that Acidithiobacillus ferrooxidans was the dominant microorganism in the lava-rock based biofilters. A novel mechanism for H2S removal in a lava-rock-based biofilter is proposed based on the biogeochemical analysis of lava rock. PMID- 15816682 TI - Specific resistance to filtration of biomass from membrane bioreactor reactor and activated sludge: effects of exocellular polymeric substances and dispersed microorganisms. AB - This study investigates the effect of dispersed microorganisms and exocellular polymeric substances on biomass dewaterability. Specific resistance to filtration (SRF) was measured for biomass from a membrane bioreactor and a completely mixed activated sludge system. Both laboratory-scale reactors were fed with synthetic wastewater and operated at a high food-to-microorganism ratio (F/M) (1 to 11 kilograms chemical oxygen demand per kilogram mixed liquor volatile suspended solids per day [kgCOD/(kg MLVSS.d)]) and short solids retention times (0.25 to 5 d). The SRF values were affected by strong interactions of three parameters: (1) the mixed liquor suspended solids concentration, (2) the amount of dispersed microorganisms, and (3) the exocellular polymeric substances (EPS) concentration. At F/M smaller than 2 kg COD/(kg MLSS.d) and mixed liquor suspended solids (MLSS) concentration higher than 2000 mg/L, increasing amount of dispersed microorganisms in the biomass yielded higher SRF values. However, at high F/M (> 5 kg COD/kg MLSS.d) and low MLSS concentrations (< 600 mg/L), lower EPS concentrations resulted in slightly smaller SRF values, even though the amount of dispersed microorganisms in the biomass was much higher. Thus, at low MLSS concentrations, EPS concentrations rather than the amount of dispersed microorganisms tend to control SRF. PMID- 15816683 TI - Inhibition of sulfide generation in a reclaimed wastewater pipe by nitrate dosage and denitrification kinetics. AB - Sulfide generation should be avoided during wastewater transportation. The efficiency of nitrate dosing for the inhibition of sulfide generation was evaluated during reclaimed wastewater transport with two nitrate doses, 2.5 and 5 mg/L nitrate-nitrogen (NO3-N). A calcium nitrate [Ca(NO3)2] solution was injected at the beginning of the 61-km-long gravity pipe, which is part of the Reclaimed Wastewater Reuse System of South Tenerife (Spain). During transportation, after dissolved oxygen depletion, a denitrification process took place. With the 5 mg/L NO3-N dose, nitrate was not completely removed at the end of the pipe, whereas with 2.5 mg/L NO3-N, a complete denitrification was achieved. Sulfide generation was completely inhibited with the 5 mg/L dose. However, with 2.5 mg/L, sulfide generation was not completely inhibited but delayed and minimized to a great extent. Denitrification was stoichiometrically limited by the availability in biodegradable matter. An empirical equation enables one to predict the nitrate concentration. PMID- 15816684 TI - A strategy for controlling deposition of struvite in municipal wastewater treatment plants. AB - This paper presents strategies to reduce the risk of struvite deposition by controlling its location of formation. Two technical routes were investigated: (1) to fix the phosphate into the dewatered sludge cake, and (2) to remove phosphate from centrate or filtrate. Chemicals used include magnesium hydroxide [Mg(OH)2], both of reagent grade and reclaimed from a flue gas desulfurization system, magnesium chloride (MgCl2), calcium hydroxide [Ca(OH)2], ferric chloride (FeCl3) and aluminum sulfate [Al2(SO4)3]. Research results indicate that (1) for anaerobically well-digested sludge, Mg(OH)2 is effective in fixing phosphate into sludge cake and improving sludge dewaterability, and (2) adding Mg(OH)2 into a reactor, located between the sludge dewatering facilities and the centrate or filtrate discharge line, and using air for mixing and carbon dioxide stripping, proves feasible in reducing struvite deposition in centrate or filtrate discharge lines and can generate a potentially valuable plant fertilizer--struvite. PMID- 15816685 TI - Nocturnal enuresis. PMID- 15816686 TI - Bed wetting in school children of Karachi. AB - OBJECTIVES: To estimate the frequency of primary nocturnal enuresis (PNE) in Pakistani children and to examine the factors associated with it. METHODS: A randomly selected cross-sectional study was conducted in five elementary schools, one in each of five districts of Karachi. The parents of 5000 children age between 3-13 years were asked to complete a questionnaire which included items about the frequency of daytime wetting and nocturnal enuresis, family history, urinary tract infection, parents and child's own concern about this problem and acquisition of treatments. RESULTS: Over all corrected response rate to the questionnaire was 69% (3395). Enuresis was present in 9.1% (308). There were 166 (53.9%) boys and 142 (46%) girls with a median age of 7 years. Only 54% (166) children sought help for their problem of which 26% consulted doctors, 16% visited homeopaths while 11% used hakeems and home remedies. Of the bed wetters, 30% were wet every night, 30% for more than three nights a week and 40% for less than three nights every week. Parents of 68.5% (211) children reported concern for the problem while 69.8% (215) children were also anxious about their enuresis. Among the concerned children group, 22% parents were not concerned about their child's problem. Family history of enuresis was present in 25.6% (79) children. CONCLUSION: The frequency of enuresis among the school going children in Karachi is 9.1% and is similar to that reported in European countries and other Asian countries including Korea and Taiwan. Enuresis causes concern to both parents and children, but only a small percentage of parents seek medical help for this problem. PMID- 15816687 TI - Non-traumatic aortic emergencies--experience from a tertiary care centre in Karachi, Pakistan. AB - OBJECTIVE: To review the incidence, clinical presentation and outcome of Non traumatic Aortic emergencies in a tertiary care hospital and its evaluation in the Emergency department (ED). METHODS: We conducted a retrospective review of cases presented to the ED at Aga Khan University Hospital during 15 year period (1988 - 2002) who had final diagnosis of Aortic Dissection or Ruptured Aortic Aneurysm. Patients without confirmatory investigations were excluded. We aimed at looking for the incidence, clinical presentation, evaluation in the ED and final outcome. RESULTS: Of the 12 cases, 7 had aortic dissection while the remaining 5 had ruptured aortic aneurysm. For Aortic dissection, mean age of presentation was 53 years with male predominance. Most of these patients had chest pain. Most common comorbid condition was hypertension. Pulse deficit was found in 2 cases, murmur in 4 cases, and focal neurologic deficit in 2 cases. Electrocardiogram revealed ischemic changes in 3 cases. Widened mediastinum on chest x-ray was present in all cases. The only initial misdiagnosis was cardiac ischemia. One patient survived to discharge. For patients presenting with ruptured aortic aneurysm, mean age of presentation was 52 yrs with a male predominance. The associated comorbid condition was hypertension. Almost all patients presented classically with abdominal pain, hypotension and palpable mass. No patient survived to discharge. CONCLUSION: Aortic emergencies although rare, are associated with significant mortality. High index of suspicion and prompt recognition by the emergency physician is of key importance. PMID- 15816688 TI - Cutaneous tuberculosis: a three year prospective study. AB - OBJECTIVE: To study the pattern of clinical presentation of cutaneous tuberculosis. METHODS: All patients with clinical suspicion of cutaneous tuberculosis, visiting outpatient department of Dermatology Unit, Jinnah Postgraduate Medical Centre, Karachi were included in this study. Total number of the patients enrolled were 74. The lesions were then classified into four clinical types viz: lupus vulgaris, scrofuloderma, tuberculids and tuberculosis verricosa cutis. RESULTS: A total of 0.1% patients visiting out patient department had cutaneous tuberculosis. Scrofuloderma was the commonest form seen in 48 (64.9%) patients, followed by lupus vulgaris in 16 (21.5%), tuberculosis verrucosa cutis in 6 (8.1%) and tuberculids 4(5.6 %). The presence of regional lymphadenopathy had correlation with the disseminated disease. CONCLUSION: There is an increasing trend of cutaneous Tuberculosis over the years. Scrofuloderma was the most common clinical presentation, followed by lupus vulgaris, tuberculosis verrucsa cutis and tuberculids. Tuberculous gumma was not seen in any patient. PMID- 15816689 TI - Evaluation of serum transferrin receptor concentration in diagnosing and differentiating iron deficiency anaemia from anaemia of chronic disorders. AB - OBJECTIVE: To diagnose and differentiate iron deficiency anaemia (IDA) from anaemia of chronic disorders (ACD) using serum concentration of soluble transferrin receptors (sTfR). METHODS: One hundred and seventy six adult anaemic patients were diagnosed on bone marrow examination as IDA and ACD in the Department of Haematology, Armed Forces Institute of Pathology, Rawalpindi from November 2001 to May 2003. They were further evaluated with sTfR, serum iron, total iron binding capacity (TIBC) and serum ferritin. These biochemical investigations were compared with results of bone marrow iron status, which served as gold standard. Absence of stainable iron in the bone marrow was diagnostic of iron deficiency, whereas abundance of iron along with decreased siderocytes and sideroblasts was considered diagnostic of ACD. Data was collected on a proforma and analysed using software SPSS (version 11.0) and t-test was used to test the statistical significance. Specificity, sensitivity positive and negative predictive value of the sTfR test was calculated. RESULTS: Out of 176 patients studied, 90 (51.1%) were diagnosed as ACD whereas 86 (48.8%) as IDA. The mean +/- SD sTfR levels in IDA patients was 9.68 +/- 2.48 mg/I, whereas mean +/- SD sTfR levels in ACD patients was 2.96 +/- 1.28 mg/I, thus clearly separating the two categories of anaemic patients. Both the sensitivity and specificity of sTfR in IDA was found to be 100%, whereas in ACD, these were 66.6% and 100% respectively. The positive and negative predictive value, in case of IDA was 100%, whereas in ACD it was 100% and 74.1% respectively. The results of serum iron, TIBC and serum ferritin correlated well in IDA, with a fall in serum iron, raised TIBC and decreased serum ferritin, except in few cases in which concomitant inflammatory conditions resulted in falsely high serum ferritin level. Serum iron and TIBC were not useful in cases of ACD. However, the serum ferritin cutoff level of 90 ng/ml was evaluated which virtually excludes IDA, and found this highly effective in cases of IDA alongwith chronic inflammatory conditions. CONCLUSION: The results show that in case of simple IDA, sTfR concentration is significantly raised and it has a very high test efficiency in this condition. However in case of ACD the positive predictive value is high (100%) but the negative predictive value is compromised (74.1%). It is therefore a reliable laboratory index of IDA and in distinguishing IDA from ACD). PMID- 15816690 TI - Adherence of private practitioners with the National Tuberculosis Treatment Guidelines in Pakistan: a survey report. AB - OBJECTIVE: In Pakistan, over 80% of the patients suffering from TB consult a private practitioner for the initial evaluation. A cross-sectional survey was conducted in seven thickly populated urban communities of Rawalpindi district to evaluate the adherence of private practitioners with TB treatment guidelines as laid down by National Tuberculosis Control Programme (NTP) in Pakistan. The data was collected over 30 days. METHODS: A young lean man was simulated to act as a TB patient and was provided with a chest X-ray suggestive of TB and two Acid-Fast Bacilli (AFB) positive sputum reports. Only those prescriptions were included for analysis which either had recognized the patient having TB or had any TB drug written in the prescription. RESULTS: A total of 77 practitioners were visited. Prescriptions of 53 general practitioners fulfilled the inclusion criteria and were analyzed. Only 2 (3.7%) prescriptions out of 53 met the required standard for TB patients as laid down by NTP. Eighty three percent (n = 44) favored a combination drug for the treatment while the rest preferred individual preparations. CONCLUSIONS: The study reflects the lack of knowledge about standardized TB treatment protocols amongst the private practitioners in Pakistan. Public Private Partnerships between government public health departments and non-governmental organizations working in public health can be a valuable tool in generating mass awareness campaigns. PMID- 15816691 TI - Benign prostatic hyperplasia: mode of presentation and postoperative outcome. AB - OBJECTIVE: To find out various modes of benign prostate hyperplasia (BPH) presentation in our region and their relation to postoperative failure to void after transurethral resection of prostate (TURP). METHODS: A study was conducted at Urology Department, Sandeman Teaching Hospital Quetta and Akram Hospital (Private) Quetta from January 2000 to December 2003. All BPH patients in whom the primary mode of presentation was accurately determined and later on underwent TURP were included in the study. Four modes of presentation were defined: (1) lower urinary tract symptoms (LUTS), (2) acute retention, (3) chronic retention and (4) and acute on chronic retention. After relevant investigations all these patients underwent TURP. Postoperatively catheter was removed when the urine was clear, usually within 48 hours of operation. Patients failing to void were recatheterized and given a second trial without catheter (TWOC) at third day of recatheterization. Resumption of spontaneous voiding on either the first or second TWOC was defined as "successful TWOC". Failure to void on second TWOC was defined as "failure to void" and was managed by a six week period of catheterization, followed by an additional TWOC. Statistical analysis was used to see any significant relation of failure to void postoperatively to mode of presentation of BPH, age of the patients and weight of the resected prostatic tissues. RESULTS: A total of 345 BPH patients were included in the study. Of these 270 (78.3%) patients presented with urinary retention and 75 (21.7%) with lower urinary tract symptoms (LUTS). Patients who presented with retention were acute retention 129 (37.4%), chronic retention 81(23.5%) and acute on chronic retention 60 (17.4%). The proportion of men failing to void after TURP was significantly higher (P < 0.05) in those with (i) acute retention as compared to LUTS (ii) chronic retention compared to acute retention and (iii) acute on chronic retention as compared to acute retention. The proportion of men failing to void postoperatively was highly significant (P < 0.005) in those with retention of any type as compared to LUTS. Age of the patients and weight of the resected prostatic tissues were found not significant factors in relation to failure to void postoperatively. CONCLUSION: BPH patients in our region present very late, most of them (> 78%) with complication of urinary retention. Mode of presentation of BPH greatly influences the postoperative outcome of this disease. Patients presenting with complications of chronic and acute on chronic retention have less favourable results regarding pOstoperative voiding after TURP. Moreover age of the patient and weight of the prostate are not significant factors in relation to failure to void postoperatively. PMID- 15816692 TI - Usefulness of needle biopsy in the diagnosis of paediatric liver disorders. AB - OBJECTIVE: To see the usefulness of liver biopsy in the diagnosis and to document the spectrum of paediatric liver diseases. METHODS: A retrospective, cross sectional survey at Histopathology Department of Army Medical College Rawalpindi from January 2000 to December 2003. The liver biopsies were taken with Menghini needle. The fixed tissues were processed under standard conditions. RESULTS: During four years period, a total of 100 cases with age range of 1.5 months to 16 years were studied. The most common histological findings in order of frequency were secondary haemochromatosis (30%), biliary atresia (20%), storage disorders (16%), cirrhosis of liver (10%) and neonatal hepatitis (10%). The less common entities were chronic hepatitis (6%), nonspecific reactive hepatitis (3%) and granulomatous hepatitis (1%). One case each of hepatoblastoma, haemophagocytic lymphohistiocytosis and congenital fibrosis was also noted. These findings have been compared with local and international histological studies. CONCLUSION: Liver biopsy is a useful diagnostic technique in the diagnosis of paediatric liver diseases. Biliary atresia, strorage disorders and neonatal hepatitis are the most common entities in our set up. PMID- 15816693 TI - Disaster drill at a university hospital. AB - The course of a disaster drill held on 23 October 2001 at Aga Khan University, Karachi is reported. The Hospital Emergency Plan was put to trial on that day. Volunteers were invited to become simulated casualties in the drill. Briefing seminars had been conducted with the key players of the hospital. The scenario was a man-made type disaster. A 747 jumbo jet with 200 passengers had crashed at the end of the runway at Quaid-e-Azam International Airport while taking off in a thunderstorm. Fifty casualties were sent to Emergency Room by ambulance. The Plan was activated and relevant units were mobilized according to the Plan. It took 2 hours to complete the disaster drill. Major difficulties were identified in the operations, communications, staff deployment, and emergency control center. Debriefing sessions reviewed difficulties encountered throughout the drill and the possible remedies. PMID- 15816694 TI - Development of a comprehensive epilepsy surgery programme in Pakistan. PMID- 15816695 TI - Solid and cystic epithelial neoplasm of pancreas with metastasis: report of a highly unusual case. AB - Solid and cystic papillary epithelial neoplasms of the pancreas are uncommon tumors occurring predominantly in young women. These tumors have excellent prognosis and after complete surgical resection, more that 95% patients are cured. Occasionally, they invade the surrounding pancreatic parenchyma. These tumors can recur even many years after resection, so long-term follow up is essential. Very few cases metastasize. We present a case of a middle aged woman with metastases to liver and omentum. PMID- 15816696 TI - Menke's kinky hair syndrome--a rare medical condition. AB - The case of a 16-month-old boy is described who had typical clinical and radiological features and was proven biochemically to be a case of Menke's disease. Clinical manifestations began in the first few months with hypothermia, hypotonia, seizures and death occurring at the age of 18 months. PMID- 15816697 TI - Extra hepatic portal vein obstruction leading to variceal bleed due to portal hypertension. AB - Portal Hypertension can be due to many causes other than cirrhosis. We report a case of extra hepatic portal vein obstruction leading to portal hypertension and varices, managed successfully by creating a Porto Caval shunt. PMID- 15816698 TI - Reasons for the problems faced by patients in government hospitals: results of a survey in a government hospital in Karachi, Pakistan. AB - OBJECTIVE: To assess the reasons for the problems faced by patients in the government hospitals to make an emphasis on higher authorities for identification of problems and implementation of effective measures. METHODS: A questionnaire based doctor's survey was conducted at Jinnah Postgraduate Medical Centre (JPMC). The survey included 10 doctors each from 5 wards of JPMC. Informed consent was taken from study participants and ethical considerations were fulfilled. All the doctors were asked questions related to the problems created by hospital management, due to the irresponsibility and concerns of the doctors and also due to the patients themselves. The responses were evaluated separately. RESULTS: A total of 50 doctors participated, out of which, 19 were males and 31 females. Out of all the reasons listed, 84% doctors favored the reason of inadequate salary to doctors & paramedical staff by government while other two reasons favored by 80% doctors were lack of facilities and security for doctors and illiteracy and poverty of patients. CONCLUSION: This hospital based study showed that the causes were related mainly to hospital management, doctors' attitude and responsibility and also to patients' illiteracy and poverty. This calls for adequate measures by higher authorities to rectify the situation. PMID- 15816699 TI - Fifteen (or so) things we love about dentistry. PMID- 15816700 TI - Report finds claims against amalgam lack scientific evidence. PMID- 15816701 TI - Musculoskeletal pain and the dental health care community. PMID- 15816702 TI - Prevalence of upper extremity symptoms and disorders among dental and dental hygiene students. AB - CONTEXT: Upper extremity musculoskeletal disorders are common among dental professionals. The natural history of these disorders is not well-understood. These disorders are more common in older workers, but the prevalence among younger workers has not been well-studied. OBJECTIVE: The objective of this study was to determine if dental/dental hygiene students had a similar prevalence of upper extremity musculoskeletal disorders compared to age-matched clerical workers. We hypothesize students will have a lower prevalence of upper extremity musculoskeletal disorders compared to clerical workers. DESIGN: This was a cross sectional design. SETTING: Dental and dental hygiene students from three schools were compared to clerical workers from three locations (an insurance company and two data processing plants). SUBJECTS: There were 343 dental and dental hygiene students and 164 age-matched clerical workers. MAIN OUTCOME MEASURES: Regional discomfort was the primary outcome. The secondary health outcomes were diagnoses of carpal tunnel syndrome and upper extremity tendinitis. RESULTS: Clerical workers had a higher prevalence of hand symptoms (62 percent vs. 20 percent), elbow symptoms (34 percent vs. 6 percent) and shoulder/neck symptoms (48 percent vs. 16 percent) and a higher prevalence of carpal tunnel syndrome (2.5 percent vs. .6 percent) and upper extremity tendinitis (12 percent vs. 5 percent). The clerical workers were more obese, smoked more, exercised less frequently, and had lower educational levels and less control of their work environment. CONCLUSIONS: Dental and dental hygiene students have a very low prevalence of upper extremity musculoskeletal disorders. A longitudinal study is necessary to evaluate ergonomic and personal risk factors. PMID- 15816703 TI - Working postures of dentists and dental hygienists. AB - A joint study was conducted by a manufacturer of dental stools in the Midwest of the United States and Marquette University to measure the occupational postures of dentists and dental hygienists. The postures of 10 dentists and 10 dental hygienists were assessed using work sampling and video techniques. Postural data of the neck, shoulders and lower back were recorded from video and categorized into 30-degree intervals: o (neutral posture of respective joint), 30, 60 and 90 degrees. Each subject's postures were observed while they were treating patients during a four-hour period, during which 100 observations of postures were recorded at random times. Compared to standing, dentists and dental hygienists were seated 78 percent and 66 percent of the time, respectively. Dentists and dental hygienists flexed their trunk at least 30 degrees more than 50 percent of the time. They flexed their neck at least 30 degrees 85 percent of the time during the four-hour duration, and their shoulders were elevated to the side of their trunk (abducted) at least 30 degrees more half of the time. The postures of the trunk, shoulders, and neck were primarily static. This database of postures can be used by dental professionals and ergonomists to assess the risk dentists and dental hygienists are exposed to musculoskeletal disorders, such as low back pain or shoulder tenosynovitis, from deviated joint postures. They could use these data to select dental furniture or dental devices that promote good body posture, i.e., reduce the magnitude and duration of deviated joint postures, which, in theory, would decrease the risk of musculoskeletal disorders. PMID- 15816704 TI - Functional training for dentistry: an exercise prescription for dental health care personnel. AB - Athlete's Performance, an organization of specialists in the development of athleticism and injury prevention, has analyzed the seated postural demands of dental health care workers for the purpose of developing an exercise protocol appropriate to the dental profession. As with their individualized exercise prescriptions for some of the world's most acclaimed athletes, the conditioning of the torso is the focus of a prescription for exercise when injury prevention is emphasized. An analysis of the seated postural demands common to dental health care workers is the basis for an exercise protocol intended to strengthen the torso and encourage "good" seated posture. PMID- 15816705 TI - Getting your back back to work: pain relief--where to start? AB - Dental health care workers are vulnerable to back and neck pain resulting from poor occupational posture. While numerous choices exist for treatment, this article will provide them with a practical approach to seeking out appropriate care for this common malady. The McKenzie treatment approach is discussed and recommendations for its application are presented to provide the reader with a starting point for treatment. For the dental health care worker experiencing pain and dysfunction of the back and/or neck, as more than half will during their careers, this article will seek to provide an overview of potential causes while creating a roadmap for seeking the most appropriate conservative "antidote" for their care. PMID- 15816706 TI - Waiting room blues. PMID- 15816707 TI - [Concerning nurses]. PMID- 15816708 TI - [The liberal struggle to create order]. PMID- 15816709 TI - [Europe against cancer]. PMID- 15816710 TI - [Meeting the need for essential pediatric care]. PMID- 15816711 TI - [Urgent vital signs in the first month]. PMID- 15816712 TI - [Assembly of implantable catheters]. PMID- 15816713 TI - [Sexuality and nursing]. PMID- 15816714 TI - [Avian influenza, principles of prevention or general distraction?]. PMID- 15816715 TI - [Organizing a protocol of ventilator weaning]. PMID- 15816716 TI - [Cold]. PMID- 15816717 TI - [Bosentan]. PMID- 15816718 TI - [The High Authority of health: as opposed to a new government system of health?]. PMID- 15816719 TI - [Recurrence of sexually transmitted diseases (STD)]. PMID- 15816720 TI - [Nurses for patient intake and orientation]. PMID- 15816721 TI - Preparation, solid-state characterization, and computational study of a crown ether attached to a squaramide. AB - [structure: see text] Crystals of a disecondary squaramide covalently linked to a crown ether presents a great variety of inter- and intramolecular nonbonded interactions including C-H/pi contacts, C-H...O and N-H...O hydrogen bonds, and pi-pi stacking between squaramide rings. Latter interaction, the stacking between squaramide rings, can be considered as an experimental evidence for the proposed aromaticity of squaramide when it is forming hydrogen bonds, either as acceptor or donor. PMID- 15816722 TI - Selective and unusual fluoride ion complexation by a steroidal receptor using OH...F- and CH...F- interactions: a new motif for anion coordination? AB - [structure: see text] A novel cholaphane has been synthesized from a naturally occurring bile acid in just two steps. It has an ability to bind two fluoride ions selectively utilizing the glycolate motif in chloroform. This "inside-out" cyclodextrin analogue encapsulates fluoride through O-H...F(-) and C-H...F(-) interaction. PMID- 15816723 TI - W(CO)5(L)-catalyzed formal cope rearrangement of allenyl silyl enol ethers. AB - [reaction: see text] On treatment of 5-siloxyhexa-1,2,5-trienes with a catalytic amount of W(CO)(6) under photoirradiation, formal Cope rearrangement proceeded to give 2-siloxyhex-1-en-5-ynes in good yield. The electrophilic activation of the allenyl moiety by W(CO)(5) triggers the intramolecular attack of the silyl enol ether in a 6-endo manner to produce a cyclohexenyl tungsten species. Carbon carbon bond cleavage occurs by electron donation from the anionic W(CO)(5) into the silyloxonium moiety to afford the products with regeneration of the W(CO)(5)(L). PMID- 15816724 TI - Asymmetric hydrogenation of isobutyrophenone using a [(Diphosphine) RuCl2 (1,4 diamine)] catalyst. AB - [reaction: see text] The use of three chiral 1,4-diamines in the [(diphosphine) RuCl(2) (diamine)] catalyst system is demonstrated in the hydrogenation of acetophenone. The use of a 1,4-diamine offers unique properties that allow tuning of the catalyst system. These include the first example of the use of a racemic diamine in combination with a chiral phosphine, which gives 95% ee in the hydrogenation of isobutyrophenone. PMID- 15816725 TI - Enantioselective H-atom transfer reaction: a strategy to synthesize formaldehyde aldol products. AB - [reaction: see text] Enantioselective radical alkylation of Baylis-Hillman adducts furnished aldol products in good yield and selectivity. The results illustrate that the selectivity in the hydrogen atom transfer is dependent on the size of the ester substituent, with smaller substituents providing better enantioselectivity. PMID- 15816726 TI - Downfield proton chemical shifts are not reliable aromaticity indicators. AB - [structure: see text] The downfield chemical shifts of arene hydrogens (delta (1)H) are due only in part to the pi ring current contribution [sigma(//)(pi)]; local framework effects are equally important. Neither proton chemical shifts nor even sigma(||)(pi) tensor elements, per se, are reliable aromaticity indicators. Unsaturated polycyclic hydrocarbons with nonaromatic quinoid structures have delta (1)H and sigma(//)(pi) values in the "aromatic range". Conversely, numerous aromatic protons, including those in five-membered ring heterocycles, resonate in the "nonaromatic range". PMID- 15816727 TI - Novel photochromic spiroheterocyclic molecules via oxidation of 1,8 diaminonaphthalene. AB - [reaction: see text] Novel spiroheterocyclic molecules, namely, 2,3-dihydro-2 spiro-7'-[8'-imino-7',8'-dihydronaphthalen-1'-amine]perimidine, PNI, and 2,3 dihydro-2-spiro-4'-[8'-aminonaphthalen-1'(4H)-one]perimidine, PNO-p, were obtained by oxidation of 1,8-diaminonaphthalene using manganese dioxide. These molecules exhibit thermally reversible photochromism with good photofatigue resistance, and their photogenerated forms possess broad absorption in the visible region. PMID- 15816729 TI - Trimethylsilyl-directed 1,3-dipolar cycloaddition reactions in the solid-phase synthesis of 1,2,3-triazoles. AB - [reaction: see text] A regioselective method for the preparation of 1,5 trisubstituted 1H-1,2,3-triazoles via a 1,3-dipolar cycloaddition of 1 trimethylsilylacetylenes with organoazides is described. Immobilization of the azide on REM resin and subsequent cycloaddition afforded a 2 x 2 x 4 x 3 membered 1,5-disubstituted 1H-1,2,3-triazole library with an average purified yield of 68%. PMID- 15816728 TI - New general sulfinylating process for asymmetric synthesis of enantiopure sulfinates and sulfoxides. AB - [reaction: see text] A general process for the efficient synthesis of sulfinyl transfer agents has been developed using cinchona alkaloids quinine and quinidine as chiral auxiliaries. The importance of these new and unique sulfinyl transfer agents is exemplified by the expedient synthesis of several sulfoxides in excellent enantiopurities and high yields. PMID- 15816730 TI - A convenient, general synthesis of 1,1-dimethylallyl esters as protecting groups for carboxylic acids. AB - [reaction: see text] Carboxylic acids were converted in high yield to their 1,1 dimethylallyl (DMA) esters in two steps. Palladium-catalyzed deprotection of DMA esters was shown to be compatible with tert-butyl, benzyl, and Fmoc protecting groups, and Fmoc deprotection could be carried out selectively in the presence of DMA esters. DMA esters were also shown to be resistant to nucleophilic attack, suggesting that they will serve as alternatives to tert-butyl esters when acidic deprotection conditions need to be avoided. PMID- 15816731 TI - First enantiospecific synthesis of the antitumor marine sponge metabolite (-)-15 oxopuupehenol from (-)-sclareol. AB - [reaction: see text] A new route toward puupehenone-related bioactive metabolites from (-)-sclareol, based on the palladium(II)-mediated diastereoselective cyclization of a drimenylphenol, is described. Utilizing this, the first enantiospecific synthesis of the antitumor and antimalarial (-)-15-oxopuupehenol, together with improved syntheses of (+)-puupehenone, (+)-puupehedione, and (+)-15 cyanopuupehenone, were accomplished. PMID- 15816732 TI - A room-temperature protocol for the rhodium(I)-catalyzed addition of arylboron compounds to sulfinimines. AB - [reaction: see text] The addition of organoboronic acids to chiral sulfinimines proceeds under mild conditions at room temperature, using Rh(I) catalysis in the absence of external phosphine ligands. Clean reaction only occurs in the presence of water as a cosolvent. The sulfinamide adducts are formed with high diastereoselectivities, providing a convenient route to the synthesis of enantiomerically enriched chiral benzylic amines. PMID- 15816733 TI - Synthesis of alpha-L-threofuranosyl nucleoside triphosphates (tNTPs). AB - [structure: see text] The alpha-l-threofuranosyl nucleoside triphosphates of T, G, and D (tTTP, tGTP, and tDTP) were synthesized from the described 2'-O-DMT protected derivatives using the Eckstein method, while the corresponding C derivative (tCTP) was prepared from the 2'-O-acetyl derivative. The prepared alpha-l-threofuranosyl nucleoside triphosphates, despite being one carbon shorter than the native 2'-deoxyfuranosyl nucleoside triphosphates, are effective substrates for selected DNA polymerases. PMID- 15816734 TI - A Pauson-Khand approach to the synthesis of ingenol. AB - [reaction: see text] Pauson-Khand cyclization of dioxanone photoadduct 21 leads to the formation of a single product in good yield. However, retro-aldol fragmentation of the pentacyclic cyclopentenone 22 leads to the formation of 23, with cis C-8/C-10 intrabridgehead stereochemistry, unlike the target compound ingenol 1, which possesses C-8/C-10 trans intrabridgehead stereochemistry. PMID- 15816735 TI - Photochemical generation of ribose abasic sites in RNA oligonucleotides. AB - [reaction: see text] A general method for the photochemical generation of ribose abasic sites within RNA oligonucleotides is reported. Photochemically caged nucleoside phosphoramidite analogues were prepared and incorporated into RNA oligonucleotides by automated RNA synthesis. Irradiation of the modified RNA at 350 nm efficiently produced ribose abasic sites at specific sites within RNA sequences. The current approach offers a chemical route to RNA abasic lesions for RNA biochemical studies. PMID- 15816736 TI - Synthesis of N,N-Dialkyl-N'-arylhydrazines via palladium-catalyzed N-arylation by using N,N-dialkylhydrazines/2LiCl adducts. AB - [reaction: see text] The reaction of N,N-dialkylhydrazine/2LiCl adducts with aryl bromides in the presence of Pd(2)(dba)(3) as the palladium source, Xantphos or X phos as the ligands, toluene as the solvent, and NaOBu-t as the base provides an efficient route to N,N-dialkyl-N'-arylhydrazines. Best results were obtained by using N,N-dialkylhydrazine/2LiCl adducts prepared in situ, omitting their isolation. PMID- 15816737 TI - Enantioselective [2 + 2] photodimerization reactions of coumarins in solution. AB - [reaction: see text] The [2 + 2]photodimerization reactions of coumarin to an optically active anti-head-to-head dimer in the presence of an optically active host compound in cyclohexane solution proceeded with high enantioselectivity. PMID- 15816738 TI - Synthesis, photonic characteristics, and mesomorphism of an oligo biphenylene vinylene pi-electron system. AB - [reaction: see text] The synthesis and photonic and liquid-crystalline properties of a novel oligo biphenylene vinylene (OBV) chromophore with an extended pi electron system are reported; the compound exhibits high fluorescence, a large two-photon absorption cross-section, and two- and three-dimensional liquid crystalline mesophases. PMID- 15816739 TI - Planar tetracoordinate carbons in cyclic hydrocarbons. AB - [structure: see text] A series of cyclic hydrocarbons containing a planar tetracoordinate carbon atom is proposed. To rationalize the electronic factors contributing to the stability of these molecules, an analysis of the molecular orbitals and the induced magnetic field is presented. PMID- 15816740 TI - Diversifying vancomycin via chemoenzymatic strategies. AB - [reaction: see text] The rapid diversification of glycopeptides via glycorandomization reveals that significantly diverse substitutions are tolerated and suggests there may be a synergistic benefit to the construction of mechanistically related natural product core scaffold fusions. This work also further highlights the utility of chemoenzymatic approaches to diversify complex natural product architectures. PMID- 15816741 TI - Application of microwave irradiation to the synthesis of 14-helical beta peptides. AB - [structure: see text] We have evaluated the effects of microwave irradiation on the solid-phase synthesis of beta-peptides. Sequences designed to adopt the 14 helix, especially those containing the structure-promoting residue trans-2 aminocyclohexanecarboxylic acid (ACHC), suffer from poor synthetic efficiency under standard SPPS conditions. A comparison of microwave and conventional heating shows that both provide excellent synthetic results for shorter sequences; however, we identify a clear benefit from microwave irradiation for longer beta-peptides. PMID- 15816742 TI - Expedient synthesis and design strategies for new peptoid construction. AB - [reaction: see text] A range of peptoids can be prepared efficiently using microwave-assisted solid-phase chemistry in a commercial reactor. This method is most effective for the installation of electronically deactivated benzylic amines. The systematic incorporation of these amines into peptoids can deliver oligomers capable of displaying unique and stable structural motifs-microwave assisted solid-phase synthesis will enable their future study and application. PMID- 15816743 TI - Preparation of 2- and 4-arylmethyl N-substituted and N,N-disubstituted anilines via a "green", multicomponent reaction. AB - [reaction: see text] A new, green, regioselective, one-step, multicomponent reaction of an aldehyde possessing a nonenolizable carbonyl function, cyclohex-2 enone (or a derivative thereof), and primary or secondary amines afforded 2-N substituted arylmethyl anilines or 4-N,N-disubstituted arylmethyl anilines, respectively. Yields and regioselectivities were good. Evidence for a pathway involving imine and iminium intermediates is presented along with examples demonstrating amenability of the process to combinatorial chemistry. PMID- 15816744 TI - Stereocontrolled [4+2]-annulation accessing dihydropyrans: synthesis of the C1a C10 fragment of kendomycin. AB - [reaction: see text] Development of new organosilane reagents bearing C-centered chirality where the stereocenter is fully substituted, and their use in the stereocontrolled synthesis of cis- and trans-dihydropyrans containing a trisubstituted olefin is described. The reagents participate in Lewis acid promoted [4+2]-annulations providing useful levels of selectivity with both aliphatic and aromatic aldehydes. A stereoselective synthesis of the C1a-C10 fragment of kendomycin (1) is also described. PMID- 15816745 TI - Stereoselective synthesis of decarestrictine D from a previously inaccessible (2Z,4E)-alkadienyl alcohol precursor. AB - [reaction: see text] The core structure of decarestrictine D was constructed by stereoselective oxygenation of (2Z,4E)-alkadienyl alcohol, which could be synthesized by a nickel-catalyzed coupling reaction between the corresponding cis bromide and trans borate. Efficiency in macrocyclization of the seco acid with Yamaguchi reagent was found to be protective-group-dependent, and the best yield of 40% was obtained with the seco acid with tri-MOM protective groups. PMID- 15816746 TI - Rhodium-catalyzed reaction of thiols with polychloroalkanes in the presence of triethylamine. AB - [reaction: see text] RhCl(PPh(3))(3) catalyzes a reaction of thiols with polychloroalkanes in the presence of triethylamine. This reaction serves as a convenient new method to produce formaldehyde dithioacetals, ethylenedithioethers, thioformates, and dithiocarbonic esters under mild conditions. PMID- 15816747 TI - Synthesis of tricyclic 4-chloro-pyrimido[4,5-b][1,4]benzodiazepines. AB - [reaction: see text] A novel methodology was developed for the efficient synthesis of 4-chloro-pyrimido[4,5-b][1,4]benzodiazepines. The key is the intramolecular Friedel-Crafts cyclization of 5-amino-4-(N-substituted)anilino-6 chloropyrimidine with either a carboxylic acid or its derivatives to construct the 4-chloro-pyrimido[4,5-b][1,4]benzodiazepine core. Subsequent nucleophilic substitution allows the introduction of one more diversity point in the target molecules. This strategy provides an efficient method to access a library of compounds based on privileged substructures that are of great interest in drug discovery. PMID- 15816748 TI - New benzo[b]furans as electroluminescent materials for emitting blue light. AB - [structure: see text] New functionalized mono- and bis-benzo[b]furan derivatives were synthesized and developed as blue-light emitting materials. They possessed a CN, CHO, CH=CHPh, CH=CPh(2), or CH=CHCOOH group at the C4-position. Two benzo[b]furan nuclei in bis-benzo[b]furan derivatives were connected by a divinylbenzene bridge. With good volatility and thermal stability, bis benzo[b]furan 7a was fabricated as a device. It emitted blue light with brightness 53430 cd/m(2) (at 15.5 V) and high maximum external quantum efficiency 3.75% (at 11 V). PMID- 15816749 TI - Efficient methods for the synthesis of 2-hydroxyphenazine based on the Pd catalyzed N-arylation of aryl bromides. AB - [reaction: see text] Substituted diphenylamines can be synthesized by Pd(0) catalyzed N-arylation using o-nitroanilines and nitro-substituted aryl bromides for a substrate. Cyclization of the diphenylamines by various methods, including the intramolecular Pd(0)-catalyzed N-arylation, produces 2-methoxyphenazine which can easily be deprotected to give 2-hydroxyphenazine. This phenazine is required to synthesize methanophenazine, a novel redoxactive cofactor isolated from methanogenic archaea. PMID- 15816750 TI - A chelation effect on the pathway between intramolecular hydrodimerization and pinacol coupling. AB - [structure: see text] Depending upon the reaction conditions, the reductive cyclization affords either the pinacol or normal hydrodimerization-type products. This selectivity is highly dependent upon the substitution at the beta-position of the enones. PMID- 15816751 TI - Highly enantioselective synthesis of (R)-alpha-alkylserines via phase-transfer catalytic alkylation of o-biphenyl-2-oxazoline-4-carboxylic acid tert-butyl ester using cinchona-derived catalysts. AB - [reaction: see text] A highly enantioselective synthetic method for (R)-alpha alkylserines was developed by the phase-transfer catalytic alkylation of o biphenyl-2-oxazoline-4-carboxylic acid tert-butyl ester (4i) using cinchona derived phase-transfer catalyst N(1)-(9-anthracenylmethyl)-O(9)-allyl dihydrocinchonidinium bromide (up to 96% ee). PMID- 15816752 TI - Synthesis and cytotoxicity studies of artemisinin derivatives containing lipophilic alkyl carbon chains. AB - [reaction: see text] Cytotoxic artemisinin derivatives have been synthesized by a modular approach of "artemisinin + linker + lipophilic alkyl carbon chain". A strong correlation between the length of the carbon chains and the cytotoxicities against human hepatocellular carcinoma (HepG2) was revealed. Notably, compared with artemisinin (IC(50) = 97 microM), up to 200-fold more potent cytotoxicity (IC(50) = 0.46 microM) could be achieved by attachment of a C(14)H(29) carbon chain to artemisinin via an amide linker. PMID- 15816753 TI - 2-Alkylidenesulfol-3-enes by (regio- and) stereoselective cheletropic addition of SO2 to (di)vinylallenes. AB - [reaction: see text] The cheletropic addition of SO(2) to divinylallenes is regioselective, taking place at the most substituted vinylallene and at the E unit if vinyl groups of opposite geometry are competing. Diastereofacial differentiation results from the approach of the reagent from the less substituted direction of the allene and from the concomitant disrotatory movement of the termini of the vinylallene to afford the sterically more congested 2 alkylidenesulfol-3-ene isomer. DFT computations confirm the regio- and the stereoselectivity of these cheletropic additions. PMID- 15816754 TI - 3'-immobilized probes with 2'-caps: synthesis of oligonucleotides with 2'-N methyl-2'-(anthraquinone carboxamido)uridine residues. AB - [reaction: see text] A synthesis for oligodeoxynucleotides with a 3'-terminal 2' N-methyl-2'-acylamido-2'-deoxyuridine residue was developed. Unlike their unmethylated counterparts, these oligodeoxynucleotides can be stably immobilized on aldehyde-displaying glass surfaces to provide DNA microarrays. An anthraquinone carboxamido group as a 2'-substituent doubled the capture efficiency of an immobilized tetradecamer. PMID- 15816755 TI - Attachment of electron-withdrawing 2,4-dinitrobenzene groups to a cryptand-based receptor for Cu(II)/H+ specific exciplex and monomer emissions. AB - [reaction: see text] In a cryptand-based fluorescent signaling system with the configuration "fluorophore-spacer-receptor", attachment of the electron withdrawing 2,4-dinitrobenzene groups to the cryptand receptor renders it highly selective for Cu(II). The system exhibits dual monomer and exciplex emissions in the presence of either of Cu(II) and H(+) as input with concomitant movements of one of the 2,4-dinitrobenzene groups. PMID- 15816756 TI - Direct amino acid-catalyzed asymmetric desymmetrization of meso-compounds: tandem aminoxylation/O-N bond heterolysis reactions. AB - [reaction: see text] A practical organocatalytic process for the synthesis of optically active, highly substituted alpha-hydroxy-ketones was achieved through asymmetric desymmetrization (ADS) of prochiral ketones. The ADS and O-N bond reduction reaction of prochiral ketone with nitrosobenzene in the presence of a catalytic amount of chiral amine or amino acid produced the tandem ADS/O-N bond reduced products as single diastereomers with good yields and excellent enantiomeric excesses. PMID- 15816757 TI - Synthesis of the putative structure of tridachiahydropyrone. AB - [reaction: see text] A short total synthesis of the putative structure of the marine natural product tridachiahydropyrone as a single enantiomer is described. Novel steps include a cuprate addition and cyclization to form a cyclohexanone ring and formation of the bicyclic pyrone with P(2)O(5) on Celite. The spectroscopic data obtained for compound 1 do not match those reported for tridachiahydropyrone; therefore, revision of the assigned natural product structure is warranted. PMID- 15816758 TI - Synthesis of silylallene glycosides and diene diglycosides by C-glycosidation of D-glucal with 1,4-bis(trimethylsilyl)-2-butyne. AB - [reaction: see text] Silylmethylallenyl glycosides, symmetrical and unsymmetrical diene glycosides, were synthesized by C-glycosidation with 1,4 bis(trimethylsilyl)-2-butyne in good yield. The nature of the product is controlled by the choice of Lewis acid, BF(3).OEt(2), or SnCl(4). The efficient construction of unsymmetrical diene glycosides was achieved in one pot on the basis of the order of addition of sugar starting materials. PMID- 15816759 TI - Enantioselective synthesis of the C18-C25 segment of lasonolide A by an oxonia cope prins cascade. AB - [reaction: see text] A 2-oxonia-Cope Prins cascade was developed that led to a facile and stereoselective synthesis of the C18-C25 segment of lasonolide A. The strategy nicely handles the introduction of the quaternary center in the tetrahydropyran ring, and all of the stereogenic centers in the product arise from a single stereocenter introduced in a catalytic enantioselective reaction. PMID- 15816760 TI - Synthesis of a C20-C26 segment of superstolide A by addition of a chiral allenylzinc reagent to (R)-N-boc alaninal. AB - [reaction: see text] Additions of chiral allenylzinc and indium reagents to N-Boc alaninal were examined as a possible route to a C20-C26 segment of superstolide A. Allenylzinc reagents, prepared in situ by palladiozincation of (R)- and (S)-5 pivalyloxy-3-butyn-2-ol mesylate, showed excellent reagent control to afford the anti,syn and anti,anti diastereomers as the nearly exclusive adducts. PMID- 15816761 TI - Consecutive approach to alkenes that combines radical addition of phosphorus hydrides with Horner-Wadsworth-Emmons-type reactions. AB - [reaction: see text] Addition of diethyl thiophosphite to terminal alkenes, in the presence of a radical initiator, followed by deprotonation of the phosphonothioate and reaction with a ketone, offers a concise one-pot approach to substituted alkenes. This novel method, which can incorporate alkylation or acylation steps, can be applied to the stereoselective formation of sterically hindered tri- and tetrasubstituted alkenes. PMID- 15816762 TI - Biomimetic synthesis of acid-sensitive (-)-caparrapi oxide and (+)-8-epicaparrapi oxide induced by artificial cyclases. AB - [reaction: see text] Asymmetric total syntheses of acid-sensitive (-)-caparrapi oxide (1) and (+)-8-epicaparrapi oxide (2) from farnesol (9) were achieved using Sharpless-Katsuki epoxidation and Lewis acid-assisted chiral Bronsted acid (chiral LBA)-induced polyene cyclization as key steps. Furthermore, (-)-1 could be directly synthesized from (S)-nerolidol (3) and (R)-LBA with 88% ds by reagent control which overcame substrate control, while (-)-2 was obtained from (R)-3 and (R)-LBA with >99% ds by the double-asymmetric induction. PMID- 15816763 TI - Observation of the first Heck-type cross-coupling reaction of allenes with aryl halides. Synthesis of polysubstituted 1,2-allenyl sulfones. AB - [reaction: see text] The Heck-type allenylation of aryl halides with allenes has been observed for the first time: The regioselectivity of intermolecular carbopalladation of 1,2-allenyl sulfones affording vinylic Pd intermediates is completely opposite to what was reported in the literature. PMID- 15816764 TI - Solid-phase synthesis of photochromic spiropyrans. AB - [reaction: see text] A convenient solid-phase synthesis of spiropyrans is described. A succinate linker was employed, which allows base-catalyzed cleavage of the desired photochromophore derivatized with a succinimide. Excellent yields and purities were obtained for the compounds that comprise a library of 23 spiropyrans. Opening of the succinimide ring in spiropyran could be realized under mild conditions. PMID- 15816765 TI - Synthesis of the axially substituted titanium Pc-C60 dyad with a convenient method. AB - [structure: see text] We successfully synthesized the axially substituted titanium Pc-C(60) dyad with a convenient method that improves on the traditional asymmetrical phthalocyanine routine to covalently linked phthalocyanines with other functional molecules. The intramolecular photoinduced process between phthalocyanine donor and fullerene acceptor was preliminarily studied. PMID- 15816766 TI - A concise route to the azaspirodecane moiety of halichlorine and structurally related alkaloids. AB - [reaction: see text] A tandem Michael addition-enolate alkylation followed by Dieckmann cyclization and Beckmann rearrangement provided the corresponding [5.4.0] azaspirobicyclodecane, a key intermediate in our synthetic route to the marine alkaloid halichlorine (1). PMID- 15816767 TI - Kinetic study of various phosphoramidite ligands in the iridium-catalyzed allylic substitution. AB - [reaction: see text] A comparative kinetic study of seven ligands is presented which clearly shows that a slight difference in the substitution pattern of the aryl group on the amine moiety of the ligand dramatically alters the activity of the resulting iridium catalyst. Ligand L6 shows the most impressive kinetics as well as the highest enantioselectivities. PMID- 15816768 TI - Cyclization or hydrogen migration: theoretical study and experimental evidence on the reactivities of unsaturated amidyl radicals. AB - [reaction: see text] Theoretical calculations (B3LYP/6-31G) backed up by deuterated experiments reveal that the N-substituents (R) play a crucial role in determining the reaction pathways of unsaturated amidyl radicals. With the increase of the bulkiness of N-alkyl group, the activation energy for 6-exo cyclization increases steadily, while the activation energy for 1,5-H abstraction remains almost unchanged. Therefore, cyclization occurs exclusively when R is H while 1,5-H migration occurs exclusively when R is t-Bu. PMID- 15816769 TI - An approach to the imine ring system of pinnatoxins. AB - [reaction: see text] A concise stereoselective approach to the spirobicyclic imine fragment of pinnatoxins and pteriatoxins is described. The approach relies on a tandem reaction sequence involving consecutive sigmatropic rearrangements to build the quaternary chiral center at the core of the spirobicyclic ring system. PMID- 15816770 TI - Nonparallelism between reaction rate and dienophile-catalyst affinity in catalytic enantioselective Diels-Alder reactions. AB - [reaction: see text] The above reaction is much faster with Y = CF(3)CH(2)O than with Y = CH(3)O. However, the methyl ester is a strong inhibitor of the Diels Alder reaction of the trifluoroethyl ester, since it has a higher affinity for the catalyst 1. PMID- 15816771 TI - Enantioselective organocatalytic Mukaiyama-Michael addition of silyl enol ethers to alpha,beta-unsaturated aldehydes. AB - [reaction: see text] A highly enantioselective, organocatalytic Mukaiyama-Michael addition reaction of silyl ethers and alpha,beta-unsaturated aldehydes has been developed. The process, catalyzed by MacMillan's chiral imidazolidinone, affords delta-keto aldehydes in high yields (56-87%) and high enantioselectivities (85 97% ee). Moreover, the reaction is applicable to a wide range of silyl ethers and alpha,beta-unsaturated aldehydes and, as such, provides access to a range of important synthetic building blocks. PMID- 15816772 TI - Synthesis of chiral vicinal diamines by highly diastereoselective three-component phenolic Mannich reaction: temperature-dependent stereodivergency. AB - [reaction: see text] A diastereoselective three-component synthesis of chiral o 1,2-diaminoalkyl phenols from an electron-rich phenol, an amine, and a chiral alpha-N,N-dibenzylamino aldehyde is developed. The diastereoselectivity of this phenolic Mannich reaction is temperature-dependent,and either anti or syn diastereomer can be prepared by controlling the reaction conditions. Low reaction temperature (-20 degrees C) favors the formation of anti adduct 1, whereas higher temperature (60 degrees C) under otherwise identical conditions produces mainly the syn isomer. PMID- 15816773 TI - Synthesis of a novel ceramide analogue via Tebbe methylenation and evaluation of its antiproliferative activity. AB - [reaction: see text] A new analogue of (2S,3R)-ceramide (2) with a methylene group at C4 has been synthesized from d-tartaric acid (3) by using Tebbe methylenation as the key step. Compound 2 exhibited markedly higher antiproliferative activity on mouse embryonic fibroblast (MEF) cells than natural (2S,3R,4E)-ceramide (1). PMID- 15816774 TI - Synthesis of highly substituted cyclohexenes via mixed Lewis acid-catalyzed Diels Alder reactions of highly substituted dienes and dienophiles. AB - [reaction: see text] A high-yielding method is described for the rapid synthesis of very hindered cyclohexenes by the Diels-Alder reaction of hindered silyloxy dienes and dienophiles using the mixed Lewis acid catalyst system (AlBr(3)/AlMe(3)). Thus, reaction of the hindered diene 4 with various substituted enones gave good yields of the expected cycloadducts even though both partners are quite sterically hindered. PMID- 15816775 TI - Synthesis of 3-arylpiperidines by a radical 1,4-aryl migration. AB - [reaction: see text] A route to 3-arylpiperidines, 3-arylpyridines, and 5 arylpiperidin-2-ones involving a radical 1,4-aryl migration has been explored. The sequence requires a xanthate addition to an N-allylarylsulfonamide, followed by acetylation and treatment with dilauroyl peroxide to give the 1,4-aryl transfer product, which upon acidic hydrolysis affords the desired piperidine derivative. PMID- 15816776 TI - Thioureas as ligands in the pd-catalyzed intramolecular Pauson-Khand reaction. AB - [reaction: see text] The thiourea-Pd complex was established as a novel type of catalyst in the PKR of allylpropargylamine, and the demonstrated chemistry may prove to be valuable for developing thiuorea as a ligand for the Pd-catalyzed Pauson-Khand reaction. PMID- 15816777 TI - [Rh(CO)2Cl]2-catalyzed domino reactions involving allylic substitution and subsequent carbocyclization reactions. AB - [reaction: see text] Three novel domino reaction processes have been discovered and developed that employ the regioselective and stereoselective [Rh(CO)(2)Cl](2) catalyzed alkylations of allylic trifluoroacetates with alpha-substituted sodiomalonates followed by an intramolecular Pauson-Khand annulation, a [5 + 2] cycloaddition, or a cycloisomerization. A unique aspect of the methodology is that a single catalyst is used to effect sequential transformations simply by increasing the temperature for the second reaction. PMID- 15816778 TI - Synthesis of furan-bridged 10-membered rings through [8 + 2]-cycloaddition of dienylfurans and acetylenic esters. AB - [reaction: see text] The coupling of various dienylfurans with dimethyl acetylenedicarboxylate (DMAD) has been examined. In most cases this reaction proceeds via [8 + 2]-cycloaddition to afford furan-bridged 10-membered ring systems as a single diastereomer. Dienylfuran intermediates were generated using either a chromium carbene-based method or aldol-based methods. Reaction of [8 + 2]-cycloadducts with electrophilic reagents occurred selectively at the enol ether alkene. PMID- 15816779 TI - Stereoselectivity control in the Rh(I)-catalyzed conjugate additions of aryl and alkenylboronic acids to unprotected hydroxycyclopentenones. AB - [reaction: see text] The stereoselective Rh(I)-catalyzed conjugate addition reaction of aryl and alkenylboronic acids to unprotected 2-phenyl-4 hydroxycyclopentenone is presented. The free OH group on the substrate is responsible for the stereochemistry, which is cis for arylboronic derivatives. In the case of the alkenylboronic compounds, the stereochemistry can be tuned to either a cis (bases as additives) or trans addition (CsF as additive) without the need of protecting groups. PMID- 15816781 TI - Clinical predictors of suicide in primary major depressive disorder. AB - OBJECTIVE: Follow-up studies of patients with depressive disorders have identified only a few replicable predictors of suicide and have not explored possible interactions between them. The following analysis takes advantage of a large cohort of depressed patients given detailed, structured interviews 2 decades ago. METHOD: The data set on which this analysis is based was collected between 1976 and 1990. Research personnel administered the Schedule for Affective Disorders and Schizophrenia to 785 adults who had major depressive disorder (Research Diagnostic Criteria) but who lacked other Axis I disorders. The current analysis used the National Death Index to determine mortality status as of 2003. RESULTS: One in 4 of the 134 deaths were by suicide for an overall suicide rate of 4.2%. In comparison to the remaining 752 patients, the 33 who died by suicide were more likely to have been inpatients and to have had a history of suicide attempts at the time of baseline assessment. They had also expressed more hopelessness and had higher ratings of suicidal tendency. The last of these variables was the most robust by far and, when tested with other predictors in regression analyses, was the only one to retain significance (p < .0001). No interactions between predictors emerged. As in an earlier, similar study, the suicidal tendency rating was predictive of suicides that occurred after the first year of follow-up, which suggests that suicidal tendencies comprise a trait that persists across episodes. CONCLUSION: A global rating of suicidality appears to be the single most important predictor of eventual suicide in patients with major depressive disorder. PMID- 15816782 TI - Prevalence of maternal depressive symptoms in low-income Hispanic women. AB - BACKGROUND: Maternal depression can have significant repercussions for the health and well-being of mothers and children. In primarily white middle-income populations, approximately 15% of mothers experience depression. Among ethnically and socioeconomically diverse populations, the prevalence of maternal depression has not been as well established. However, the highest rates have been observed among low-income women. Because information about minority, underserved women is particularly sparse, we utilize data from the San Mateo County, California, Prenatal to Three project to describe the prevalence and self-recognition of depressive symptoms among low-income Hispanic mothers of infants and toddlers. METHOD: Telephone interviews of a random sample of women who received Medicaid and gave birth in San Mateo County provided our sample of 218 nonpregnant Hispanic mothers. High levels of depressive symptoms were defined as a score of > or =10 on the Edinburgh Postnatal Depression Scale (EPDS). We performed descriptive analyses and analyses of variance. RESULTS: Twenty-three percent of mothers reported high levels of depressive symptoms. Half of them recognized a need for help with depression. CONCLUSIONS: High levels of maternal depressive symptoms were prevalent among the Hispanic women on Medicaid, but only half of the women experiencing these symptoms identified themselves as needing help with depression. PMID- 15816783 TI - Incidence of delirium in older adults newly prescribed lithium or valproate: a population-based cohort study. AB - BACKGROUND: The use of lithium carbonate for the treatment of mood disorders in old age has decreased at a dramatic rate in favor of valproate. Because of lithium's narrow therapeutic range, neurotoxicity can be an important complication in lithium therapy and potentially influence prescription patterns. Therefore, we compared the incidence of delirium in older adults with mood disorders who were newly dispensed either lithium or valproate. METHOD: Using 4 population-based administrative databases from the province of Ontario, Canada (the Ontario Drug Benefit program, the Canadian Institute for Health Information, the Ontario Health Insurance Plan, and the Registered Persons Data Base), we were able to identify a cohort of mood disorder patients 66 years and older who were newly dispensed lithium or valproate over an 8-year period (1993-2001). Measures were taken to ensure that the sample was composed of mood disorder patients. As a comparator, we included a known deliriogenic drug, benztropine. The main outcome measure was a new diagnosis of delirium on a hospitalization record during 1 year of follow-up. RESULTS: Our study cohort consisted of 2422 new users of lithium and 2918 new users of valproate over an 8-year period. There was no statistically significant difference in the incidence of delirium between lithium (2.8 per 100 person-years) and valproate (4.1 per 100 person-years). Compared with patients who received lithium, patients who received benztropine had a significantly higher risk of delirium (p < .001). CONCLUSION: The incidence of hospitalizations with delirium was similar in patients treated with lithium and valproate. These findings add to the evidence suggesting that the shift away from the use of lithium carbonate to manage mood disorders in older adults is not justified on the basis of concerns of neurotoxicity. PMID- 15816784 TI - Female reproductive cycle and obsessive-compulsive disorder. AB - BACKGROUND: The aim of our study was to assess whether there is a relationship between reproductive cycle events and the initiation or changes in symptoms of obsessive-compulsive disorder (OCD). METHOD: Forty-six female outpatients meeting DSM-IV criteria for OCD completed a semistructured interview at our OCD unit to assess the relationship between reproductive cycle events and OCD. Dates of data collection were from January 2001 to December 2003. RESULTS: In our sample, OCD onset occurred in the same year as menarche in 22% (N = 10), at pregnancy in 2% (N = 1), at postpartum in 7% (N = 3), and at menopause in 2% (N = 1). Worsening of preexisting OCD was reported by 20% of patients (9/45) at premenstruum, 8% (1/12) at pregnancy, 50% (6/12) at postpartum, and 8% (1/12) at menopause. The number of premenstrual mood symptoms, which included anxiety, irritability, mood lability and depressed mood, was associated with both premenstrual worsening of OCD (OR = 5.1, p < .01) and onset or worsening of OCD at postpartum (OR = 2.7, p < .05). Patients with an onset or worsening of OCD at postpartum also more frequently reported pre-menstrual worsening of OCD and previous history of major depressive disorder, including postpartum depression (p < or =.05 for all). CONCLUSION: In a substantial number of patients, the onset or worsening of OCD was related to reproductive cycle events, especially at menarche and postpartum. Certain women with OCD seem to be vulnerable to worsening of OCD at different reproductive periods that imply hormonal fluctuations, and premenstruum and post partum were the 2 reproductive events with a greater vulnerability. Those patients whose OCD symptoms appeared to be related to reproductive events also exhibited a greater history of mood symptoms (premenstrual depression and major depressive episodes). PMID- 15816785 TI - Actual driving performance and psychomotor function in healthy subjects after acute and subchronic treatment with escitalopram, mirtazapine, and placebo: a crossover trial. AB - OBJECTIVE: The effects of escitalopram 10 to 20 mg/day and mirtazapine 30 to 45 mg/day on actual driving and psychomotor performance of 18 healthy subjects were determined in a randomized, double-blind, placebo-controlled, multiple-dose, 3 way crossover trial. METHOD: Each treatment period lasted for 15 days and was separated from the next period by a washout period of at least 13 days. Subjects received an evening dose of escitalopram 10 mg, mirtazapine 30 mg, or placebo from days 1 to 7 and an evening dose of escitalopram 20 mg, mirtazapine 45 mg, or placebo from days 8 to 15. On days 2, 9, and 16, reflecting acute period, dose increase, and steady state, respectively, the Road Tracking Test was performed. The main parameter was standard deviation of lateral position. Psychomotor performance was also assessed on days 2, 9, and 16 by laboratory computer tasks. Subjective sleep quality was measured with the Groninger Sleep Quality Scale, and mood was measured by visual analogue scales. RESULTS: Treatment differences were apparent during the acute treatment period, in which subjects treated with mirtazapine 30 mg performed less well on the driving test as compared to placebo. The Divided Attention Task results also revealed a significant increase in tracking error after a single dose of mirtazapine 30 mg as compared to placebo. Mirtazapine decreased feelings of alertness and contentedness. Mirtazapine did not affect performance on days 9 and 16 of treatment. Escitalopram did not affect driving, psychomotor performance, or subjective mood throughout treatment. CONCLUSION: Driving performance, as well as psychomotor functioning, was not affected by escitalopram treatment in healthy subjects. Driving performance was significantly impaired after ingestion of mirtazapine 30 mg during the acute treatment period. PMID- 15816786 TI - Pregnancy outcome of women using atypical antipsychotic drugs: a prospective comparative study. AB - BACKGROUND: A substantial number of women of childbearing age suffer from schizophrenia and other mental illnesses that require the use of antipsychotic drugs. Atypical antipsychotics have been on the market since the mid-1990s, and to date there are no prospective comparative studies regarding use during pregnancy. OBJECTIVES: (1) To determine whether atypical antipsychotics increase the rate of major malformations above the 1% to 3% baseline risk seen in the general population. (2) To examine rates of spontaneous and therapeutic abortions, rates of stillbirths, birth weight, and gestational age at birth. METHOD: The cohort was composed of pregnant women who contacted the Motherisk Program in Canada or the Israeli Teratogen Information Service in Israel and women who were recruited from the Drug Safety Research Unit database in England. Women who had been exposed to atypical antipsychotics were matched to a comparison group of pregnant women who had not been exposed to these agents. RESULTS: Data were obtained on 151 pregnancy outcomes that included exposure to olanzapine (N= 60), risperidone (N = 49), quetiapine (N = 36), and clozapine (N = 6). Among women exposed to an atypical antipsychotic, there were 110 live births (72.8%), 22 spontaneous abortions (14.5%), 15 therapeutic abortions (9.9%), and 4 stillbirths (2.6%). Among babies of women in this group, there was 1 major malformation (0.9%), and the mean +/-SD birth weight was 3341 +/-685 g. There were no statistically significant differences in any of the pregnancy outcomes of interest between the exposed and comparison groups, with the exceptions of the rate of low birth weight, which was 10% in exposed babies compared with 2% in the comparison group (p = .05), and the rate of therapeutic abortions (p = .003). CONCLUSION: These results suggest that atypical antipsychotics do not appear to be associated with an increased risk for major malformations. PMID- 15816787 TI - Olanzapine increases slow wave sleep and sleep continuity in SSRI-resistant depressed patients. AB - OBJECTIVE: The atypical antipsychotic drug olanzapine has been employed as an augmentation treatment in depressed patients unresponsive to treatment with selective serotonin reuptake inhibitors (SSRIs). In healthy subjects, acute olanzapine administration increases sleep continuity and enhances slow wave sleep (SWS). The aim of the present study was to determine if the addition of olanzapine to SSRI treatment in depressed patients produced similar effects on sleep. METHOD: We measured the effect of open-label olanzapine addition (2.5 mg/day initially) on the polysomnograms of 12 patients referred from primary care sources who met DSM-IV criteria for major depressive disorder and who had had an unsatisfactory response to therapeutic doses of an SSRI. Patients were first enrolled in November 2001; final assessment occurred in November 2003. Sleep polysomnogram recordings were made on 3 occasions: before olanzapine addition, on the first night of olanzapine treatment, and after 3 weeks of olanzapine treatment. RESULTS: After the first night of olanzapine treatment and during the third week, subjects showed improvements in sleep efficiency (p < .001), subjective sleep quality (p < .05), and SWS (p < .01). Scores on the Hamilton Rating Scale for Depression fell significantly (p = .001), with the majority of the decrease being apparent after the first week of treatment. CONCLUSION: Olanzapine improves sleep continuity and increases SWS in patients receiving SSRI treatment. These effects are apparent after the first dose of olanzapine and are maintained for the next 3 weeks. The ability of olanzapine to increase SWS is probably attributable to 5-HT(2A/2C) receptor blockade, which has been identified as a relevant mechanism in the therapeutic effect of olanzapine in SSRI-resistant depressed patients. PMID- 15816788 TI - Psychotherapy and medication in the treatment of adult and geriatric depression: which monotherapy or combined treatment? AB - OBJECTIVE: The authors reviewed the literature with respect to the relative efficacy of medications and psychotherapy alone and in combination in the treatment of depression. DATA SOURCES AND STUDY SELECTION: Findings from empirical studies comparing medications and psychotherapy alone and in combination were synthesized and prognostic and prescriptive indices identified. We searched both MEDLINE and PsychINFO for items published from January 1980 to October 2004 using the following terms: treatment of depression, psychotherapy and depression, and pharmacotherapy and depression. Studies were selected that randomly assigned depressed patients to combined treatment versus monotherapy. DATA SYNTHESIS: Medication typically has a rapid and robust effect and can prevent symptom return so long as it is continued or maintained, but does little to reduce risk once its use is terminated. Both interpersonal psychotherapy (IPT) and cognitive-behavioral therapy (CBT) can be as effective as medications in the acute treatment of depressed outpatients. Interpersonal psychotherapy may improve interpersonal functioning, whereas CBT appears to have an enduring effect that reduces subsequent risk following treatment termination. Ongoing treatment with either IPT or CBT appears to further reduce risk. Treatment with the combination of medication and IPT or CBT retains the specific benefits of each and may enhance the probability of response over either monotherapy, especially in chronic depressions. CONCLUSION: Both medication and certain targeted psychotherapies appear to be effective in the treatment of depression. Although several prognostic indices have been identified that predict need for longer or more intensive treatment, few prescriptive indices have yet been established to select among the different treatments. Combined treatment can improve response with selected patients and enhance its breadth (IPT) or stability (CBT). PMID- 15816789 TI - A review of the evidence for the efficacy and safety of trazodone in insomnia. AB - OBJECTIVE: Trazodone, a triazolopyridine antidepressant, is currently the second most commonly prescribed agent for the treatment of insomnia due to its sedating qualities. Given trazodone's widespread use, a careful review of the literature was conducted to assess its efficacy and side effects when given for treatment of insomnia. DATA SOURCES: In April 2003, a MEDLINE search was conducted using the search terms trazodone and insomnia and trazodone and sleep and restricted to 1980-2003, human subjects, and English language. As trazodone has been implicated in cardiac disorders, a further search was conducted using the term cardiac and trazodone. STUDY SELECTION: All clinical trials that measured any endpoint for insomnia efficacy were included in the assessment. A total of 18 studies were identified from the literature search. In addition, commonly used texts were consulted for information regarding adverse effects related to trazodone. DATA EXTRACTION: Because so few studies were identified by the literature search, all were evaluated and described. DATA SYNTHESIS: Evidence for the efficacy of trazodone in treating insomnia is very limited; most studies are small, conducted in populations of depressed patients, raise issues of design, and often lack objective efficacy measures. Side effects associated with trazodone are not inconsequential, with a high incidence of discontinuation due to side effects, such as sedation, dizziness, and psychomotor impairment, which raise particular concern regarding its use in the elderly. There is also some evidence of tolerance related to use of trazodone. CONCLUSION: Given the relative absence of efficacy data in patients with insomnia and the adverse events associated with trazodone's use in general, it is uncertain whether the risk/benefit ratio warrants trazodone's use in nondepressed patients with insomnia. PMID- 15816790 TI - Recidivism in medication-noncompliant serious juvenile offenders with bipolar disorder. AB - OBJECTIVE: To determine whether the recidivism rate varies for adolescent serious juvenile offenders with bipolar disorder in response to compliance with antimanic medication. METHOD: Probation records were reviewed for all adolescents (N = 31) released during a 1-year period (April 1, 1993-March 31, 1994) from a county juvenile corrections treatment facility who had DSM-III-R bipolar disorder, were stabilized on medication, and had agreed to continue treatment at an adolescent psychiatry clinic. New offenses and probation violations committed during the 12 month period after release were tallied. These recidivism records were then compared with medical records to ascertain whether these acts were committed while subjects were on (taking) or off (not taking) medication. RESULTS: The number of serious offenses (felonies and misdemeanors) was significantly reduced while subjects were on medication (4 offenses in 2992 days) versus off medication (39 offenses in 6108 days) (p < .0001). The off-medication rate of offending was 4.8 times higher than the on-medication rate. Probation violations were also significantly reduced while subjects were on medication (p < .001). CONCLUSION: Compliance with prescribed antimanic medication can markedly decrease recidivism in serious juvenile delinquents with bipolar disorder. PMID- 15816791 TI - The relation between anger expression, depression, and somatic symptoms in depressive disorders and somatoform disorders. AB - BACKGROUND: In previous studies, the relationship between either anger suppression and depression or anger suppression and somatic symptoms was examined. However, the relationship between anger expression, depression, and somatic symptoms was not examined in depressive disorders and somatoform disorders. METHOD: The DSM-IV-diagnosed subjects included 73 patients with depressive disorders and 47 patients with somatoform disorders. The Anger Expression Scale was used to assess the level of anger expression or suppression. The severity of depression was assessed using the Symptom Checklist-90-Revised (SCL-90-R). The Somatization Rating Scale and the SCL-90-R somatization subscale were used to assess the severity of somatic symptoms. Data were collected from March 2000 to March 2001. RESULTS: The results of the path analyses showed that in depressive disorder patients, anger expression had a stronger effect on somatic symptoms through depression than did anger suppression, although both anger expression and anger suppression had a significant indirect effect on somatic symptoms. The depressive disorder group also showed a significant but negative direct effect of anger suppression on anger expression in the path from anger suppression to anger expression to depression to somatic symptoms. However, only anger suppression had an indirect effect on somatic symptoms through depression in somatoform disorder patients. CONCLUSIONS: The results suggest that anger expression might play a more predominant role in depression and somatic symptoms of depressive disorder patients than anger suppression, but only anger suppression might be associated with depression and somatic symptoms of somatoform disorder patients. In addition, incomplete anger suppression followed by anger expression is likely to be associated with depression and somatic symptoms in depressive disorders. PMID- 15816792 TI - Clinical differences between suicidal and nonsuicidal depressed children and adolescents. AB - OBJECTIVE: To examine the clinical symptoms and comorbid psychiatric disorders of depressed children and adolescents with and without clinically significant suicidal ideation. METHOD: Children and adolescents aged 7 to 17 years with current DSM-III-R major depressive disorder (MDD) (N = 135) were recruited between January 1987 and April 2002. Current MDD symptoms and lifetime comorbid psychiatric disorders were assessed using either a combination of the Schedule for Affective Disorders and Schizophrenia for School Age Children-Epidemiologic and -Present Episode versions or the -Present Lifetime version. Thirty-two percent (N = 43) of the depressed subjects were classified as suicidal (at least suicidal ideation with a plan). RESULTS: Depressed suicidal youth presented with a more severe episode (p = .001) and a poorer functional status (p = .019), were more hopeless (p = .001), and presented more frequently with insomnia (p = .011). There was an interaction between suicide x sex x pubertal status for severity of MDD (p = .013), the presence of hopelessness (p < .001), poor functional status (p = .023), and comorbidity with a lifetime history of any disruptive behavior (p = .019). Among pre-pubertal depressed males, suicidal boys had significantly increased severity of MDD (p = .025) and poorer functional status (p = .044) than non-suicidal boys. Among postpubertal depressed females, suicidal girls were more frequently hopeless (p = .008) and presented an increased severity of MDD (p = .022) and more frequent lifetime history of any disruptive behavior (p = .03) when compared with nonsuicidal girls. CONCLUSION: There appears to be a sex difference for some clinical features, particularly hopelessness, among depressed suicidal children and adolescents. Whether hopelessness is a sex-specific characteristic of depressed suicidal children and adolescents requires further study. PMID- 15816793 TI - Ondansetron treatment in Tourette's disorder: a 3-week, randomized, double-blind, placebo-controlled study. AB - OBJECTIVE: The aim of the present study was to evaluate the efficacy of ondansetron, a selective 5-HT(3) antagonist, in the treatment of Tourette's disorder. METHOD: Participants (N = 30) aged 12 to 46 years, diagnosed with DSM IV Tourette's disorder and resistant to previous haloperidol treatment, were enrolled in a 3-week, randomized, double-blind, placebo-controlled outpatient study. Assessments were conducted at baseline and once a week during the study period. Scales used included the Tourette's Syndrome Global Scale (TSGS), the Yale Global Tic Severity Scale (YGTSS), and the Yale-Brown Obsessive Compulsive Scale. Ondansetron dose was 8, 16, and 24 mg/day in the first, second, and third weeks, respectively. RESULTS: A significant positive effect of ondansetron on tic severity, as assessed by the TSGS, was noted (baseline vs. endpoint: mean +/-SD = 29.62 +/-20.33 vs. 20.58 +/-12.82, p = .002 vs. placebo). However, no significant effect was detected upon assessing ondansetron/ placebo effect on tic severity with the YGTSS (baseline vs. endpoint: mean +/-SD = 24.04 +/-9.44 vs. 17.50 +/ 9.48, p = .15 vs. placebo). No change in obsessive-compulsive symptoms was noted in either group. Adverse effects included mild and transient abdominal pain. CONCLUSIONS: Ondansetron may have antitic effects in patients with Tourette's disorder. Large-scale, double-blind studies should further assess the antitic efficacy of ondansetron. PMID- 15816794 TI - Atypical antipsychotics and glucose homeostasis. AB - OBJECTIVES: Persistent reports have linked atypical antipsychotics with diabetes, yet causative mechanisms responsible for this linkage are unclear. Goals of this review are to outline the pathogenesis of nonimmune diabetes and to survey the available literature related to why antipsychotics may lead to this disease. DATA SOURCES: We accessed the literature regarding atypical antipsychotics and glucose homeostasis using PubMed. The search included English-language publications from 1990 through October 2004. Keywords used included atypical antipsychotics plus one of the following: glucose, insulin, glucose tolerance, obesity, or diabetes. In addition, we culled information from published abstracts from several national and international scientific meetings for the years 2001 through 2004, including the American Diabetes Association, the International Congress on Schizophrenia Research, and the American College of Neuropsychopharmacology. The latter search was necessary because of the paucity of well-controlled prospective studies. STUDY SELECTION: We examined publications with significant new data or publications that contributed to the overall comprehension of the impact of atypical antipsychotics on glucose metabolism. We favored original peer-reviewed articles and were less likely to cite single case studies and/or anecdotal information. Approximately 75% of the fewer than 150 identified articles were examined and included in this review. DATA EXTRACTION: Validity of data was evaluated using the existence of peer-review status as well as our own experience with methodology described in the specific articles. DATA SYNTHESIS: The metabolic profile caused by atypical antipsychotic treatment resembles type 2 diabetes. These agents cause weight gain in treated subjects and may induce obesity in both visceral and subcutaneous depots, as occurs in diabetes. Insulin resistance, usually associated with obesity, occurs to varying degrees with different antipsychotics, although more comparative studies with direct assessment of resistance are needed. A major problem in assessing drug effects is that psychiatric disease itself can cause many of the manifestations leading to diabetes, including weight gain and sedentary lifestyle. While studies in healthy subjects are limited and inconclusive, studies in animal models are more revealing. In the conscious canine model, some atypical antipsychotics cause adiposity, including visceral obesity, a strong risk factor for the metabolic syndrome. Furthermore, while few studies have examined effects of antipsychotics on pancreatic beta-cell function, canine studies demonstrate that expected beta cell compensation for insulin resistance may be reduced or even eliminated with these agents. CONCLUSIONS: Atypical antipsychotics have been shown to contribute to weight gain, which may well reflect increased body fat deposition. Such increased fat is known to cause resistance to insulin action, although more information regarding effect on insulin action is needed. The effect of these drugs on fat distribution has been clearly shown in animal models. It is known that the normal response to insulin resistance is compensatory hyperinsulinemia, which may prevent diabetes. In animals, there is evidence that the hyperinsulinemic compensation is inadequate in the face of atypical antipsychotic agents. It remains to be examined whether failure of adequate pancreatic beta cell compensation for insulin resistance plays a central role in the pathogenesis of diabetes associated with this class of drugs. PMID- 15816795 TI - Mirtazapine for obsessive-compulsive disorder: an open trial followed by double blind discontinuation. AB - BACKGROUND: Many patients with obsessive-compulsive disorder (OCD) experience little response to standard treatment with serotonin reuptake inhibitors. Mirtazapine enhances serotonergic function by a mechanism distinct from reuptake inhibition. Because a pilot study suggested effectiveness of mirtazapine in OCD, we conducted a controlled trial. METHOD: We recruited 30 subjects, 15 treatmentnaive and 15 treatment-experienced, with DSM-IV OCD of > or =1 year's duration and a Yale-Brown Obsessive Compulsive Scale (YBOCS) score of > or =20. In the 12-week, open-label phase, subjects received mirtazapine starting at 30 mg/day and titrated over 2 weeks as tolerated to 60 mg/day. At week 12, responders (YBOCS score decrease > 25%) were randomly assigned, double-blind, to continue mirtazapine or switch to placebo for 8 weeks, including a 1-week, double blind taper week for placebo subjects. RESULTS: In the open-label phase, the mean +/-SD YBOCS score fell from 28.3 +/-3.7 to 20.3 +/-8.5 (paired samples t = 4.81, p < .0001). Four subjects (13.3%) discontinued for side effects. Sixteen subjects (53.3%) (8 treatmentnaive, 8 treatment-experienced) were responders and 15 agreed to randomization. Response was independent of comorbid mood disorders. In the 8 week, double-blind, placebo-controlled discontinuation phase, the mirtazapine group's mean YBOCS score fell a mean +/-SD of 2.6 +/-8.7 points while the placebo group's mean score rose a mean +/-SD of 9.1 +/-7.5 points (Mann Whitney U = 6.5, p = .005, 1-tailed). All other outcome measures were consistent with mirtazapine's superiority versus placebo. CONCLUSION: Mirtazapine may be an effective pharmacotherapy for OCD. If our results are replicated, larger double blind studies would be indicated. PMID- 15816796 TI - Excessive daytime sleepiness in young adults: a 20-year prospective community study. AB - OBJECTIVE: Excessive daytime sleepiness (EDS) is a symptom with high clinical and public health importance because of its association with increased risk for accidents, decreased productivity, and impaired quality of life. Little information is available regarding the longitudinal course or clinical correlates of EDS. The aim of this study was to explore associations between self-reported EDS, sleep disorder symptoms, major depression, and anxiety in a longitudinal community study of young adults. METHOD: A prospective single-age community study of young adults (Zurich Cohort Study) was conducted from 1978 through 1999. Information was derived from 6 interviews administered when participants (N = 591) were ages 20, 22, 27, 29, 34, and 40 years. Trained health professionals administered a semistructured interview for health habits and psychiatric and medical conditions. The presence of either or both of 2 symptoms-accidentally falling asleep or excessive need for sleep during the day-was used to establish the presence of EDS. RESULTS: EDS was a common complaint among the study participants, with increasing prevalence with age. Cross-sectionally, EDS was associated with insomnia symptoms, nocturnal hypersomnia, anxiety disorders, somatization, and reduced quality of life. Longitudinally, impaired sleep quality, waking up too early, and anxiety were associated with later EDS. Conversely, EDS was not significantly associated with later anxiety or depressive disorders. CONCLUSIONS: Insomnia symptoms and anxiety are associated with the subsequent occurrence of EDS. Although these findings do not demonstrate causality, insomnia and anxiety disorders are prevalent and treatable conditions, and our results may have important clinical implications for the prevention and treatment of EDS. Whether the results of this study are limited to populations with elevated levels of psychopathology remains to be tested. PMID- 15816797 TI - Suicide in schizophrenia, clozapine, and adoption of evidence-based medicine. PMID- 15816798 TI - Clozapine diminishes suicidal behavior: a retrospective evaluation of clinical records. AB - OBJECTIVE: To test the antisuicidal effect of clozapine, taking into consideration some potentially confounding variables. METHOD: A retrospective evaluation was conducted of the clinical charts of 94 inpatients treated continuously with clozapine for at least 6 weeks between 1962 and 1994. In a mirror design, a period of continuous clozapine treatment (mean duration of 15 months) was compared with a pre-clozapine period of equal length, and in 17 patients also with a post-clozapine period, with regard to suicidal behavior. The role of variables such as staying in a protective hospital milieu and receiving treatment with typical neuroleptics and antidepressants was considered. RESULTS: The rate of suicidal behavior was 28% (26/94) in the pre-clozapine period, 3% (3/94) in the clozapine period, and 18% (3/17) in the post-clozapine period, the corresponding figures for serious suicidal behavior requiring medical attention being 12% (11/94), 1% (1/94), and 12% (2/17), respectively. The odds ratios were 11.6 (95% CI = 3.4 to 39.9) and 12.3 (95% CI = 1.6 to 97.5) for suicidal and serious suicidal behavior, respectively, in favor of the clozapine period in comparison with the pre-clozapine period. Staying in the hospital was associated with reduction in suicidal behavior. The antisuicidal effect of clozapine possibly disappears at doses that are too low. CONCLUSION: Clozapine diminishes the frequency of suicidal behavior including serious suicidal acts, regardless of comedication with antidepressants. In the protective hospital milieu, this effect is less pronounced, and it disappears after clozapine discontinuation. PMID- 15816800 TI - Efficacy of quetiapine in generalized social anxiety disorder: results from an open-label study. PMID- 15816801 TI - Mirtazapine for alcohol dependence: a case report. PMID- 15816802 TI - Electroconvulsive therapy for coexistent schizophrenia and obsessive-compulsive disorder. PMID- 15816804 TI - Underpowered repetitive transcranial magnetic stimulation might not be an effective antidepressant treatment. PMID- 15816805 TI - Low-dose risperidone and quetiapine as monotherapy for comorbid anxiety and depression. PMID- 15816807 TI - ATP-binding cassette transporter G8 M429V polymorphism as a novel genetic marker of higher cholesterol absorption in hypercholesterolaemic Japanese subjects. AB - The ratio of serum plant sterols to cholesterol is positively correlated with the fractional cholesterol absorption, whereas serum precursors of cholesterol synthesis are positively correlated with cholesterol synthesis. Recently, two ABC (ATP-binding cassette) transporters, ABCG5 and ABCG8, have been described as playing an important role in the absorption and excretion of sterols. In the present study, we tested the hypothesis that genetic variation in ABCG5/ABCG8 influences the levels of serum plant sterol (sitosterol) and cholesterol precursor (lathosterol) in Japanese primary hypercholesterolaemic patients (n = 100). We identified a novel mutation [859T/C (C287R)] and a novel polymorphism [1285A/G (M429V)] at the ABCG5/ABCG8 loci, as well as four polymorphisms reported previously [1810C/G (Q604E), 161G/A (C54Y), 1199C/A (T400K) and 1895C/T (A632V)]. In carriers of the novel M429V variant, the serum level of sitosterol and the sitosterol/cholesterol ratio were significantly higher than those in non-carriers (3.64 compared with 2.56 microg/ml, and 1.45 microg/mg compared with 1.00 microg/mg respectively; P < 0.01 for both), and serum lathosterol tended to be lower (1.95 microg/ml compared with 3.03 microg/ml; P = 0.08), whereas no significant difference was observed in other lipid profiles. These four polymorphisms (1810C/G, 161G/A, 1199C/A and 1285A/G) generated six haplotypes, and the C/G/C/G haplotype was significantly associated with a higher sitosterol level and sitosterol/cholesterol ratio compared with the other five haplotypes (P < 0.05 for both). We conclude that, in 8% of patients with hypercholesterolaemia, the novel ABCG8 M429V variant was associated with higher cholesterol absorption efficiency. Future studies should investigate whether these findings have implications for the optimal cholesterol-lowering drug treatment in hypercholesterolaemic patients. PMID- 15816809 TI - Aetiology of autism: findings and questions. AB - BACKGROUND: Although there is good evidence that autism is a multifactorial disorder, an adequate understanding of the genetic and non-genetic causes has yet to be achieved. METHODS: Empirical research findings and conceptual reviews are reviewed with respect to evidence on possible causal influences. RESULTS: Much the strongest evidence concerns the importance of susceptibility genes, but such genes have yet to be identified. Specific somatic conditions (such as tuberous sclerosis and the fragile X anomaly) account for a small proportion of cases. Over recent decades there has been a major rise in the rate of diagnosed autism. The main explanation for this rise is to be found in better ascertainment and a broadening of the diagnostic concept. Nevertheless, some degree of true rise cannot be firmly excluded. However, the epidemiological evidence on the main hypothesized environmental explanation, namely the measles-mumps-rubella vaccine, is consistently negative. CONCLUSION: Progress on the elucidation of the causes of autism will be crucially dependent on the combination of epidemiology with more basic science laboratory studies. PMID- 15816810 TI - Performance of young people with Down syndrome on the Leiter-R and British picture vocabulary scales. AB - BACKGROUND: The British picture vocabulary scales (BPVS-II) and the Leiter international performance scales (Leiter-R), both restandardised in 1997, are often used in experimental studies to match individuals with intellectual impairment. Both provide a brief measure of mental age, and cover a wide ability range using a simple format. The BPVS-II assesses verbal comprehension and the Leiter nonverbal abilities. The issue is which to choose. People with Down syndrome (DS), for example, have particular problems in language and so the BPVS may provide an underestimation of ability. METHOD: The present study investigated this by comparing the performance of 46 young people with DS (21 females, 25 males, mean age 19 years 10 months) on the BPVS-II (verbal mental age - VMA) and the Leiter-R brief IQ (nonverbal mental age - NVMA). RESULTS: Contrary to expectations VMAs were significantly higher than NVMAs (6 years 6 months and 5 years 2 months, respectively). There was a significant correlation of 0.61 between the VMA and NVMA, and both discriminated participants at all levels of ability. However, the Leiter-R brief IQ scores provided poor discrimination at the bottom end of the IQ range (IQ 36). CONCLUSION: Both the BPVS-II and the Leiter-R provide mental age equivalent scores that are useful for plotting developmental progress, although absolute mental ages may differ. PMID- 15816811 TI - Cognitive, emotional, physical and social effects of growth hormone treatment in adults with Prader-Willi syndrome. AB - BACKGROUND: Prader-Willi syndrome (PWS) is a multisystem genetic disorder characterized by short stature, muscular hypotonia, hyperphagia, obesity, maladaptive behaviour, hypogonadism and partial growth hormone (GH) deficiency (GHD). Severe GHD of other aetiologies has been shown to affect mood and quality of life negatively, and there are reports of improvements with GH replacement. We have studied cognitive, emotional, physical and social parameters in PWS adults at baseline, during and after GH treatment. PATIENTS AND METHODS: Nineteen patients, 9 females and 10 males, median age 25 years, mean BMI 35 kg/m2 participated in this study. Approximately half of the group had GHD. All patients fulfilled the clinical criteria for PWS and 13 had a positive genotype. The patients were randomized to 6 months of treatment with either GH [1.6 IU/day (0.53 mg/day)] or placebo, followed by 12 months of active GH treatment. Treatment was then stopped, and the patients were followed for an additional period of 6 months. A test battery for general cognitive evaluation and a computer-based measurement of reaction time, motor speed and fluency were employed at baseline, after 6 months and at the end of GH treatment. At the same time intervals, a self-evaluation questionnaire was answered at the end of each test session. Other questionnaires reflecting the patients' cognitive, emotional, physical and social status were answered by relatives/caretakers at baseline and at 3 and 6 months following cessation of GH treatment. RESULTS: Baseline cognitive level was estimated to be moderately to mildly impaired; IQ range was 40-90. The results from some of the cognitive and the motor performance tests improved significantly after 6 and 18 months of GH treatment. According to the questionnaires, both the patients and the relatives/caretakers evaluated physical status rather negatively at baseline, but still, impairments in both physical and social status and overall functioning were observed when GH treatment was discontinued. The self-evaluation did not change in any aspect during GH treatment. CONCLUSIONS: In this pilot study of an adult PWS cohort, we were able to document beneficial effects in mental speed and flexibility and in motor performance during GH treatment. Impairment was seen in physical and social status as well as overall functioning, when GH treatment stopped. Studies of larger cohorts are needed to further elucidate the role of GH treatment in this group of patients. PMID- 15816812 TI - Assessment of patients with intellectual disability using the International Classification of Functioning, Disability and Health to evaluate dental treatment tolerability. AB - BACKGROUND: Patients with serious intellectual disability (ID) are occasionally unable to tolerate dental treatment when intravenous sedation or general anaesthesia (IVS/GA) is involved. In order to make a decision regarding the application of IVS/GA, the International Classification of Functioning, Disability and Health (ICF) is useful. Therefore, in this study, a set of codes involved in dental problems were chosen from the ICF, and patients with ID who could tolerate dental treatments were compared with those who could not. METHODS: From preliminary interviews of six patients with ID, 16 codes were chosen, and an objective five-rank scale was then constructed for use with all chosen codes. Forty-nine ID patients who visited the Okayama University Hospital for dental treatment between January and April 2003 were evaluated. Facility workers were interviewed according to the code set chosen. The participants were then divided into two subgroups depending on their tolerability of dental treatment. The results of these groups for all 16 codes were then compared. RESULTS: Of the 49 patients interviewed, 23 were able to tolerate the dental treatment. In the 'Activities & Participation' section of the ICF, the tolerable group showed lower disability levels with regard to d110 Watching, d540 Dressing and d550 Eating. In other sections, there were no significant differences between the groups. The code set chosen in this study and the five-rank scales in each code were useful as they enabled easy interviewing. CONCLUSIONS: The ICF was raised as a possibility for considering the application of IVS/GA for dental treatment on patients with ID. For clinical use of the ICF, it is recommended that significant codes should be selected and that the five-rank scale is used so that more objective results are obtained from interviews. PMID- 15816813 TI - A survey of sleep problems in autism, Asperger's disorder and typically developing children. AB - BACKGROUND: Sleep problems are common in typically developing (TD) children and in children with autism, however, less is known about the sleep of children with Asperger's disorder (AD). The aim of this study was to compare sleep patterns of children with autism and AD to a TD group of children. METHODS: Sixty-six parents of TD children, 53 parents of children with autism, and 52 parents of children with AD completed a survey on their child's sleep patterns, the nature and severity of any sleep problems and success of any treatment attempted. RESULTS: The results showed high prevalence of sleep problems with significantly more problems reported in the autism and AD groups (TD = 50%, autism = 73%, AD = 73%), with no significant differences between groups on severity or type of sleep problem. Children with AD were significantly more likely to be sluggish and disoriented after waking and had a higher Behavioral Evaluation of Disorders of Sleep (BEDS) total score compared to the other two groups. The autism and AD groups reported significantly better treatment success for medication compared to the TD group. The autism group reported significantly better success for behavioural treatment compared to the AD group. CONCLUSIONS: In conclusion, children with AD may have more symptoms of sleep disturbance, and different types of sleep problems than children with autism. As this is the first study to compare autism and AD and to survey treatment outcomes, further research is needed to validate these findings. PMID- 15816814 TI - The association between environmental events and self-injurious behaviour in Cornelia de Lange syndrome. AB - BACKGROUND: There has been limited empirical research into the environmental causes of self-injury in Cornelia de Lange syndrome. The present study examined the variability of self-injurious behaviour in Cornelia de Lange syndrome across environmental setting events. Additionally, the association between setting events and more specific environmental events was examined. METHOD: A descriptive analysis of observational data on eight children with Cornelia de Lange syndrome aged between 4 and 14 years was carried out. The association between self injurious behaviour and four environmental setting events and between specific environmental events and setting events was examined using established statistical methods for observational data. RESULTS: Seven out of eight of the participants showed at least one form of self-injurious behaviour that was associated with a particular setting event. The study also demonstrated that the relationship between setting events and environmental events is extremely variable across individuals. CONCLUSIONS: Self-injurious behaviour in some individuals with Cornelia de Lange syndrome is associated with environmental events although the precise nature of the association warrants clarification. Using broad setting events as a methodological tool in isolation provides some insight into the role of specific environmental factors in maintaining self injurious behaviour but the integrity of setting events must be established. PMID- 15816815 TI - Correlation between subjective and objective measures of outcome in staffed community housing. AB - BACKGROUND: A distinction is made between objective and subjective assessment when quality of life evaluation is considered. The aim was to explore the association between objective and subjective appraisals within similar quality of life domains. METHODS: Correlations between scores on objective and subjective quality of life measures concerning choice, activity and integration were investigated by administering measures to the residents of a random sample of 47 small community housing services. Correlations between the measures and resident adaptive behaviour were also investigated. RESULTS: All objective measures were significantly correlated with adaptive behaviour but only one subjective measure was. With level of adaptive behaviour controlled, 6/7 correlations between pairs of objective measures were significant. Fifteen of the 16 correlations between objective and subjective measures were insignificant. CONCLUSIONS: Assessments of objective life conditions and personal satisfaction appear to be distinct. Their suitability for different purposes and the notion that subjective appraisal of life may be under homeostatic regulation are discussed. PMID- 15816816 TI - Measurement of perceived competence in Dutch children with mild intellectual disabilities. AB - BACKGROUND: Little research has been conducted on the perceived competence of children with mild intellectual disabilities (MID). One of the reasons for the marked absence of research appears to be the lack of reliable and clearly valid measurement instruments for this particular group of children. In the present study, it was examined whether a pictorial scale originally designed to measure perceived competence in typically developing children could successfully be used with children with MID. METHODS: The pictorial scale was administered to a group of 106 children with MID. The construct validity, reliability and stability of the scale were investigated. RESULTS: The results of the exploratory factor analyses and the confirmatory factor analyses supported the conceptual framework proposed. The construct validity was also supported by the pattern of intercorrelations between the subscales. The scale had adequate internal consistency and the stability analyses showed sufficient stability across a 4 month period. CONCLUSIONS: The findings show the psychometric properties of the pictorial scale to justify its use with children with MID. PMID- 15816817 TI - Beating the barriers: recruitment of people with intellectual disability to participate in research. AB - BACKGROUND: The health status of people with intellectual disability (ID) is poor, yet very little research has been done on their health needs. There are several barriers to performing this research. The aim of our research was to examine the significant barriers encountered in recruiting adults with ID to participate in research. METHODS: Our project was a randomized controlled trial aiming to improve health advocacy of people with ID. We approached many organizations to recruit participants and kept records of our results. Recruitment was very low for the first 4 months. Then we adopted two new recruitment strategies--in-service telephone recruitment and meetings with prospective participants. We then monitored the subsequent recruitment rates. We also questioned participants about the difficulties they encountered when considering recruitment. RESULTS: Initial recruitment of participants yielded less than one-third of the expected number. Additional strategies implemented were partially successful. Significant barriers to recruitment included several sectoral issues and the challenges arising from the research process. CONCLUSIONS: While this population is a difficult one to reach for research, attempts to do so should not be abandoned, because the potential health benefits for this underserved group far outweigh the recruitment barriers experienced. PMID- 15816818 TI - Response to Emerson et al. (Journal of Intellectual Disability Research, 48, 201 202). PMID- 15816821 TI - Gone fishin' in 2005-insights into the inner workings of dendritic cells. PMID- 15816822 TI - Skinomics. PMID- 15816823 TI - Control of human hair growth by neurotrophins: brain-derived neurotrophic factor inhibits hair shaft elongation, induces catagen, and stimulates follicular transforming growth factor beta2 expression. AB - Neurotrophins are important modulators of epithelial-mesenchymal interactions. Previously, we had shown that brain-derived neurotrophic factor (BDNF) and its high-affinity receptor tyrosine kinase B (TrkB) are prominently involved in the control of murine hair follicle cycling. We now show that BDNF and TrkB are also expressed in the human hair follicle in a manner that is both hair cycle dependent and suggestive of epithelial-mesenchymal cross-talk between BDNF secreting dermal papilla fibroblasts of anagen hair follicles and subpopulations of TrkB+ hair follicle keratinocytes. As functional evidence for an involvement of BDNF/TrkB in human hair growth control, we show in organ-cultured human anagen hair follicles that 50 ng per mL BDNF significantly inhibit hair shaft elongation, induce premature catagen development, and inhibit keratinocyte proliferation. Quantitative real-time rtPCR analysis demonstrates upregulation of the potent catagen inducer, transforming growth factor beta2 (TGFbeta2) by BDNF, whereas catagen induction by BDNF was partially reversible through co administration of TGFbeta-neutralizing antibody. This suggests that TrkB-mediated signaling promotes the switch between anagen and catagen at least in part via upregulation of TGFbeta2. Thus, human scalp hair follicles are both a source and target of bioregulation by BDNF, which invites to target TrkB-mediated signaling for therapeutic hair growth modulation. PMID- 15816824 TI - Novel and established potassium channel openers stimulate hair growth in vitro: implications for their modes of action in hair follicles. AB - Although ATP-sensitive potassium (K(ATP)) channel openers, e.g., minoxidil and diazoxide, can induce hair growth, their mechanisms require clarification. Improved drugs are needed clinically. but the absence of a good bioassay hampers research. K(ATP) channels from various tissues contain subtypes of the regulatory sulfonylurea receptor, SUR, and pore-forming, K(+) inward rectifier subunits, Kir6.X, giving differing sensitivities to regulators. Therefore, the in vitro effects of established potassium channel openers and inhibitors (tolbutamide and glibenclamide), plus a novel, selective Kir6.2/SUR1 opener, NNC 55-0118, were assessed on deer hair follicle growth in serum-free median without streptomycin. Minoxidil (0.1-100 microM, p<0.001), NNC 55-0118 (1 mM, p<0.01; 0.1, 10, 100 microM, p<0.001), and diazoxide (10 microM, p<0.01) increased growth. Tolbutamide (1 mM) inhibited growth (p<0.001) and abolished the effect of 10 microM minoxidil, diazoxide and NNC 55-0118; glibenclamide (10 microM) had no effect, but prevented stimulation by 10 microM minoxidil. Phenol red stimulated growth (p<0.001), but channel modulator responses remained unaltered. Thus, deer follicles offer a practical, ethically advantageous in vitro bioassay that reflects clinical responses in vivo. The results indicate direct actions of K(ATP) channel modulators within hair follicles via two types of channels, with SUR 1 and SUR 2, probably SUR2B, sulfonylurea receptors. PMID- 15816825 TI - A multicenter study of the pharmacokinetics of tacrolimus ointment after first and repeated application to children with atopic dermatitis. AB - The pharmacokinetics of tacrolimus after first and repeated application of 0.1% tacrolimus ointment were evaluated in 39 children, aged 6-12 y, with moderate to severe atopic dermatitis. The patients were grouped according to the size of the affected body surface area to be treated: Group 1< or =1500 cm(2); Group 2 >1500 cm(2) < or =3000 cm(2); Group 3 >3000 cm(2) < or =5000 cm(2). Serial blood samples to calculate pharmacokinetic parameters taken on Day 1 (first ointment application) and Day 14 (last application) showed minimal systemic exposure to tacrolimus. Overall, 92% of the blood samples assayed contained tacrolimus concentrations below 1 ng per mL and 17% of samples were below 0.025 ng per mL, the lower limit of quantification. Systemic exposure to tacrolimus varied between patients and tended to increase proportionally as the size of the treated body surface area increased. Absorption decreased with time as the skin lesions healed and there was no evidence of systemic accumulation. The mean apparent half-life of tacrolimus (t(1/2, z)) was 66+/-27 h (range 19-125 h). Most patients experienced substantial clinical improvement in their atopic dermatitis. There were no clinically relevant changes in laboratory values, and the most frequently reported adverse event was skin burning, which resolved quickly as the skin condition improved. PMID- 15816826 TI - Epidemiology of staphylococcal scalded skin syndrome in Germany. AB - Staphylococcal scalded skin syndrome (SSSS) is a blistering skin disorder caused by Staphylococcus aureus. The "Dokumentationszentrum schwerer Hautreaktionen", a unique population-based registry for severe skin reactions, included SSSS during a time period of 2 y in Germany. Statistical calculations indicated a low overall incidence between 0.09 and 0.13 cases per 1 million inhabitants per year with 95% confidence interval of [0-4]. The age distribution showed two clusters; one in young children and one in adults. The mortality rate was much lower in children than in adults. Young age was the main risk factor, whereas immunosuppression and consumptive infectious disease were the detected risk factors in adults. PMID- 15816827 TI - Global gene expression analysis of keloid fibroblasts in response to electron beam irradiation reveals the involvement of interleukin-6 pathway. AB - Keloid is a dermal fibroproliferative lesion of unknown etiology that commonly recurs after surgical excision. Post-operative adjuvant electron beam (EB) irradiation has been successfully used to reduce keloid recurrences. To provide new insights into the molecular mechanism behind the effect of EB irradiation, we used a cDNA microarray screening of more than 5000 genes to assess early changes in gene expression between EB-irradiated and non-irradiated keloid and non lesional dermal fibroblasts. Primary fibroblast cultures from keloid and associated non-lesional dermis obtained from five patients were exposed to 15 Gy EB irradiation and analyzed after 15 min incubation. Early response to EB irradiation showed that 96 (1.8%) genes were modulated 2-fold or more in keloid fibroblasts. Upregulated genes accounted for 29.2% (28 genes), whereas downregulated genes comprised 70.8% (68 genes), indicating a silencing of many genes in keloid fibroblasts after EB irradiation. Many of the downregulated genes play roles in the enhancement of cell proliferation and extracellular matrix production, whereas several of the upregulated genes involves in the promotion of apoptosis and extracellular matrix (ECM) degradation. Using emerging bioinformatic tools and further corroboration, the interleukin 6 (IL-6) signaling pathway was found to be mainly involved in EB irradiation response. We also showed co-expression of IL-6 and its specific receptor (IL-6Ralpha) in keloid fibroblasts that points to the existence of an IL-6 autocrine loop in these cells. These results suggested that at the molecular level, EB irradiation might hinder keloid formation by regularizing disturbances in the homeostatic equilibrium between inducer and inhibitor activities in the matrix system most likely through the IL-6 pathway. Our study provides clues for the molecular mechanism(s) behind the beneficial effect of EB irradiation in reducing keloid recurrences and may help develop alternative strategies for the therapy and prophylaxis of this lesion. PMID- 15816828 TI - A lack of Birbeck granules in Langerhans cells is associated with a naturally occurring point mutation in the human Langerin gene. AB - A heterozygous mutation in the Langerin gene corresponding to position 837 in the Langerin mRNA was identified in a person deficient in Birbeck granules (BG). This mutation results in an amino acid replacement of tryptophan by arginine at position 264 in the carbohydrate recognition domain of the Langerine protein. Expression of mutated Langerin in human fibroblasts induces tubular-like structures that are negative for BG-specific antibodies and do not resemble the characteristic structural features of BG. PMID- 15816829 TI - Differential activation profiles of multiple transcription factors during dendritic cell maturation. AB - Immature dendritic cells (DC) at the environmental interfaces, such as the skin, constantly survey the tissue for the emergence of microbial products and pro inflammatory mediators. Upon recognition of such "danger" signals, they undergo dynamic reprogramming of gene expression and functions, the process known as DC maturation, which plays critical roles in both innate and adaptive immune responses. Although DC have been shown to discriminate different maturation stimuli by expressing stimulus-specific signature genes and unique phenotypic and functional properties, underlying mechanisms for this extraordinary plasticity remain relatively unclear. We hypothesized that DC might activate unique sets of transcription factors (TF) upon sensing different stimuli. To test this hypothesis, we transduced a mouse epidermal-derived DC line XS106 to express the luciferase reporter gene under the control of each of 15 different cis-enhancer elements. The resulting DC panels were then exposed to 14 different microbial, endogenous, environmental, and pharmacological agents that produced unique maturational changes. This approach allowed systematic determination of TF activation profiles in DC. Our results revealed striking diversity, with different classes of stimuli triggering preferential activation of distinct sets of TF. We propose that differential TF usage represents a previously unrecognized mechanism regulating the direction of DC maturation. PMID- 15816830 TI - Loxosceles sphingomyelinase induces complement-dependent dermonecrosis, neutrophil infiltration, and endogenous gelatinase expression. AB - Envenomation by the spider Loxosceles can result in dermonecrosis and severe ulceration. Our aim was to investigate the role of the complement system and of the endogenous metalloproteinases in the initiation of the pathology of dermonecrosis. Histological analysis of skin of rabbits injected with Loxosceles intermedia venom and purified or recombinant sphingomyelinases showed a large influx of neutrophils, concomitant with dissociation of the collagenous fibers in the dermis. Decomplementation, using cobra venom factor, largely prevented the influx of neutrophils, while influx of neutrophils was also reduced in genetically C6-deficient rabbits, suggesting roles for both C5a and the membrane attack complex in the induction of dermonecrosis. However, C-depletion and C6 deficiency did not prevent the haemorrhage and the collagen injury. Zymography analysis of skin extracts showed the induction of expression of the endogenous gelatinase MMP-9 in the skin of envenomated animals. Rabbit neutrophils contained high levels of MMP-9, expression of which was further increased after incubation with venom, suggesting that these cells may be a source of the MMP-9 found in the skin of envenomated animals. Furthermore, skin fibroblasts also secreted MMP-9 and MMP-2 upon incubation with venom, suggesting that locally produced MMPs can also contribute to proteolytic tissue destruction. PMID- 15816831 TI - Detection of laminin 5-specific auto-antibodies in mucous membrane and bullous pemphigoid sera by ELISA. AB - Mucous membrane pemphigoid (MMP) is an autoimmune bullous disease that primarily affects mucous membranes leading to a scarring phenotype. MMP patients produce auto-antibodies (auto-ab) that preferentially recognize two components of the dermoepidermal basement membrane zone (BMZ): bullous pemphigoid (BP)180 and laminin 5 (LN5). Since detection of disease-specific auto-ab may be critical for diagnosis of MMP, we developed an ELISA with affinity-purified native human LN5. A total of 24 MMP, 72 BP, and 51 control sera were analyzed for LN5-specific auto ab: 18/24 (75.0%) MMP and 29/72 (40.3%) BP sera were LN5 reactive. Sensitivity and specificity of the LN5 ELISA for MMP were 75% and 84.3%, respectively, and 40.3% and 88.2% for BP, respectively. The LN5 ELISA was more sensitive than a dot blot assay with native LN5, which detected LN5-reactive IgG in 14/24 (58.3%) MMP and 16/72 (22.2%) BP sera. In MMP, but not BP, levels of LN5-reactive IgG correlated with disease severity. Furthermore, IgG reactivity to LN5 of the MMP and BP sera was not significantly associated with IgG reactivity against other autoantigens of the BMZ, such as BP180 or BP230. Thus, the established LN5 ELISA holds great promise as a novel diagnostic and prognostic parameter for MMP. PMID- 15816832 TI - Resistance to activation-induced cell death and bystander cytotoxicity via the Fas/Fas ligand pathway are implicated in the pathogenesis of cutaneous T cell lymphomas. AB - By engaging Fas, Fas ligand (FasL) on activated T lymphocytes induces activation induced cell death (AICD), and also triggers apoptosis of target cells during immune downregulation. We previously showed that within cutaneous T cell lymphoma (CTCL) lesions, malignant CD4(+) T cells expressing FasL accumulated, and were inversely distributed with CD8(+) T cells. We thus determined the responses of human CTCL cells to AICD and their cytotoxic to Fas(+) target T cells in vitro. CTCL cells expressing Fas were resistant to AICD following activation by CD3 monoclonal antibody (mAb) whereas still undergoing apoptosis if Fas was ligated to Fas mAb. CTCL cell lines, as well as Sezary Syndrome patients' peripheral blood lymphocytes, exhibited ratio-dependent cytotoxicity to Fas(+) Jurkat cells. The kinetic study showed that FasL surface expression was absent before activation, and its expression was low and/or delayed after activation. We therefore hypothesize that CTCL cells express functional FasL possibly contributing to bystander cytotoxicity within tumor infiltrates. In addition, decreased and/or delayed FasL surface expression following activation may in part contribute to their resistance to AICD. Both bystander cytotoxicity and resistance to AICD are likely to contribute to the loss of cytotoxic anti-tumor CD8(+) T cells as well as the accumulation of malignant T cells in CTCL. PMID- 15816833 TI - Corticotropin-releasing hormone (CRH) downregulates interleukin-18 expression in human HaCaT keratinocytes by activation of p38 mitogen-activated protein kinase (MAPK) pathway. AB - It is generally accepted that corticotropin-releasing hormone (CRH) acts as the main coordinator of the central response to stress. Stress or an abnormal response to stressors has been found to modify the evolution of skin disorders, including psoriasis and atopic dermatitis. Nevertheless, the specific pathogenic role of stress remains unknown in skin diseases. Interleukin (IL)-18, a member of the IL-1 family, is a key mediator of peripheral inflammation and host defense responses, and is secreted by human keratinocytes. Here, we investigated the regulatory effect of CRH on expression of IL-18 in skin keratinocytes. Exposure of HaCaT cells to CRH resulted in a reduction of IL-18 mRNA transcripts and its production was in a concentration-dependent manner. In order to investigate whether the mitogen-activated protein kinase (MAPK) signaling pathway is involved in the downregulation of IL-18 production, cells were pre-treated with SB203580, an inhibitor of p38 MAPK, prior to the addition of CRH. This pre-treatment blocked the decrease in IL-18 production. In addition, CRH treatment induced rapid phosphorylation of p38 MAPK. SB203580 were able to inhibit CRH-induced p38 MAPK phosphorylation. CRH also inhibited production of IL-18 in human primary keratinocytes. These results suggest that CRH regulates IL-18 production through the MAPK signaling pathway in human keratinocytes. PMID- 15816834 TI - Characterization of multiple P2X receptors in cultured normal human epidermal keratinocytes. AB - ATP-gated ion channels (P2X) are expressed in human epidermis and cultured keratinocytes. The aim of this study was to characterize native P2X receptors in normal human epidermal keratinocytes (NHEK) using whole-cell patch clamp technique, RT-PCR, and determination of intracellular Ca(2+) concentration ([Ca(2+)](i)). Application of ATP resulted in an inward current with a reversal potential of 0 mV. Response to ATP showed two types of currents: the slowly desensitizing response and the rapidly desensitizing response. The slowly desensitizing response was blocked by iso-pyridocaphosphate-6-azophenyl-2', 5' disulfonic acid (PPADS), a P2X receptor antagonist. We found that the expression of multiple P2X(2), P2X(3), P2X(5), and P2X(7) receptor subtype mRNA was increased in differentiated cells. On the other hand, the expression of G-protein coupled P2Y(2) mRNA was downregulated in differentiated cells. Increases in [Ca(2+)](i) evoked by alphabeta-methylene ATP (alphabeta-meATP) and 2', 3'-O-(4 benzoylbenzoyl) ATP (BzATP) were elevated, whereas elevation of [Ca(2+)](i) evoked by uridine 5'-triphosphate (UTP) was decreased in differentiated cells. Application of ATP or UVB radiation increased the expression of P2X(1), P2X(2), P2X(3), and P2X(7) receptors in NHEK. Changes in the expression levels and cation influx via multiple P2X receptors might be involved in the regulation of differentiation and one of the epidermal external sensors. PMID- 15816835 TI - Nanostructure of the epidermal extracellular space as observed by cryo-electron microscopy of vitreous sections of human skin. AB - The newly developed method, cryo-electron microscopy of vitreous sections, was used to observe the nanostructure of the epidermal extracellular space. The data were obtained from vitreous sections of freshly taken, fully hydrated, non-cryo protected human skin. The extracellular space of viable epidermis contains desmosomes, expressing a characteristic extracellular transverse approximately 5 nm periodicity, interconnected by a relatively electron lucent inter-desmosomal space. The extracellular space between viable and cornified epidermis contains transition desmosomes at different stages of reorganization interconnected by widened areas expressing a rich variety of complex membrane-like structures. The extracellular space of cornified epidermis contains approximately 9, approximately 14, approximately 25, approximately 33, approximately 39, approximately 44, and approximately 48 nm thick regions in turn containing one, two, four, six, eight, eight, and ten parallel electron-dense lines, respectively, between adjacent corneocyte lipid envelopes. The eight-line approximately 44 nm thick regions are most prevalent. PMID- 15816836 TI - Transcriptional profiling of keratinocytes reveals a vitamin D-regulated epidermal differentiation network. AB - 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] regulates mineral homeostasis and exhibits potent anti-proliferative, prodifferentiative, and immunomodulatory activities. It mediates these effects by binding to the vitamin D receptor (VDR), which belongs to the superfamily of steroid/thyroid hormone nuclear receptors. As a result of keratinocyte differentiation and anti-proliferation activities, 1,25(OH)(2)D(3) and its synthetic analogs are therapeutically effective in psoriasis and show promise for the treatment of actinic keratosis and squamous cell carcinoma. To elucidate the VDR signaling pathway in keratinocytes, we examined the gene expression profile with 1,25(OH)(2)D(3) treatment using oligonucleotide microarrays. Out of the 12,600 genes investigated, 82 were upregulated and 16 were downregulated and many of these were involved in differentiation, proliferation, and immune response. We have identified three vitamin D-responsive chromosomal loci (1p36, 19q13, and 6p25) and show the induction of various class II tumor suppressor/growth-regulatory genes in response to 1,25(OH)(2)D(3). Finally, quantitative differences in gene expression revealed a vitamin D-regulated differentiation network and identified peptidylarginine deiminases, kallikreins, serine proteinase inhibitor family members, Kruppel-like factor 4, and c-fos as vitamin D-responsive genes, whose protein products may play an important role in epidermal differentiation in normal and diseased state. PMID- 15816837 TI - Interleukin-4 suppresses the enhancement of ceramide synthesis and cutaneous permeability barrier functions induced by tumor necrosis factor-alpha and interferon-gamma in human epidermis. AB - Ceramide is an integral part of the extracellular lipid bilayer of the stratum corneum (SC) that forms the permeability barrier of the skin. The production of SC ceramides is catalyzed by sphingomyelinase (SMase) and glucocerebrosidase (GCase). Acid-ceramidase (acid-CDase) catalyzes the hydrolysis of ceramide in the SC. We examined the effects of T helper (Th)1 and Th2 cytokines on levels of transcripts of genes for acid-CDase, acid-SMase, and GCase, on levels of ceramide, and on the extent of transepidermal water loss (TEWL) in the human epidermis in an effort to determine whether these cytokines affect the permeability barrier functions. Levels of transcripts for acid-SMase and GCase and the amount of ceramide in human epidermal sheets were enhanced by tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma and these effects were inhibited in the presence of interleukin (IL)-4. In epidermal keratinocytes cultured under submerged conditions, however, no similar inhibitory effects of IL 4 were observed. Consistent with these results, TEWL was suppressed by TNF-alpha and IFN-gamma, and these effects were also inhibited by IL-4. The balance between Th1 and Th2 might affect the construction and/or the repair of the epidermal permeability barrier via regulation of the production of ceramide. PMID- 15816838 TI - Keratinocytes act as a source of reactive oxygen species by transferring hydrogen peroxide to melanocytes. AB - Basal hydrogen peroxide (H(2)O(2)) levels in normal human epidermal keratinocytes (NHEK) and melanocytes (mel) were compared on a per cell basis and found to be significantly higher in keratinocytes. Since H(2)O(2) is a neutral molecule capable of permeating through cellular membranes, we then investigated the possibility that H(2)O(2) transfer might occur between these two types of cells. Because the ratio of keratinocytes to mel in skin is 36:1, keratinocytes may act as a source of reactive oxygen species (ROS) even by passive diffusion and, thus, affect melanocytic functions. In order to measure H(2)O(2) transfer, a fluorescence-based co-culture system was developed in which mel were first pre labeled with 5-(and-6)-chloromethyl-2',7'-dichlorodihydro-fluorescein diacetate (DCFdA). When mel were co-cultured with keratinocytes, fluorescence increased as a function of keratinocyte cell number. Thus, for mel incubated with 1-, 1.5-, and 2-fold the number of keratinocytes, fluorescence increased by 22.6% (+/ 2.8%), 25.6% (+/-4.8%), and 39.9% (+/-4.1%), respectively. Separating the cells with a transwell membrane did not prevent the transfer, whereas the addition of catalase to media significantly reduced the transfer of H(2)O(2) to mel. In conclusion, keratinocytes appear to be a previously unexamined source of ROS that may affect neighboring skin cells, such as mel, and, as a result, may influence the process of melanogenesis or contribute to the progression of vitiliginous lesions. PMID- 15816840 TI - Melanoma cells express elevated levels of phosphorylated histone H2AX foci. AB - When human cells sustain a DNA double-strand break (dsb), histone H2AX in chromatin surrounding the DNA break is phosphorylated, marking repair foci. The number of phosphorylated histone H2AX (gammaH2AX) foci approximates the number of dsb present in the cell's nuclear DNA. We observed 0.4 gammaH2AX foci per nucleus in primary human melanocytes. In contrast, in four melanoma cell lines, we detected 7-17 gammaH2AX foci per nucleus, a 17-42 times increase in the basal level of gammaH2AX foci in melanoma cells relative to melanocytes (MC). Thus, untreated melanoma cells express significantly greater numbers of gammaH2AX foci than do untreated MC. Detection and rejoining of ionizing radiation-induced DNA dsb proceeded as rapidly in melanoma cells as in MC. Melanoma cells, however, reduced the number of radiation-induced gammaH2AX foci down only to pre irradiation levels. Co-localization of the majority of gammaH2AX foci with ataxia telangiectasia mutated, BRCA1, 53BP1, and Nbs1 foci in untreated melanoma cells indicated that the additional foci in melanoma cells were associated with a DNA change that the cells interpret as DNA dsb. Co-localization of gammaH2AX foci with the telomere replication factor 1 protein in untreated melanoma cells indicates that the additional foci in untreated melanoma cells are associated with dysfunctional telomeres that induce a DNA damage stress response. PMID- 15816839 TI - 4-Tertiary butyl phenol exposure sensitizes human melanocytes to dendritic cell mediated killing: relevance to vitiligo. AB - The trigger initiating an autoimmune response against melanocytes in vitiligo remains unclear. Patients frequently experience stress to the skin prior to depigmentation. 4-tertiary butyl phenol (4-TBP) was used as a model compound to study the effects of stress on melanocytes. Heat shock protein (HSP)70 generated and secreted in response to 4-TBP was quantified. The protective potential of stress proteins generated following 4-TBP exposure was examined. It was studied whether HSP70 favors dendritic cell (DC) effector functions as well. Melanocytes were more sensitive to 4-TBP than fibroblasts, and HSP70 generated in response to 4-TBP exposure was partially released into the medium by immortalized vitiligo melanocyte cell line PIG3V. Stress protein HSP70 in turn induced membrane tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression and activation of DC effector functions towards stressed melanocytes. Melanocytes exposed to 4-TBP demonstrated elevated TRAIL death receptor expression. DC effector functions were partially inhibited by blocking antibodies to TRAIL. TRAIL expression and infiltration by CD11c+ cells was abundant in perilesional vitiligo skin. Stressed melanocytes may mediate DC activation through release of HSP70, and DC effector functions appear to play a previously unappreciated role in progressive vitiligo. PMID- 15816841 TI - Proteomic profiling identifies an UV-induced activation of cofilin-1 and destrin in human epidermis. AB - The human skin is the only line of defense against UV radiation. A series of responses to protect the skin are induced by UV radiation. In this study, a proteomic approach was used to study these responses. We have performed high resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis of (solar simulated) UV-exposed reconstructed skin equivalents as well as native skin. Differentially expressed proteins were processed for mass spectrometric analysis, when consistent differences were observed in all individual human skin equivalents. In addition to proteins known to be involved in UV responses (HSP27, MnSOD, and PDX-2), we identified two novel proteins that were downregulated following UV exposure. Further analysis revealed that these proteins were the phosphorylated forms of the actin cytoskeleton modulators cofilin-1 and destrin. The de-phosphorylation of cofilin-1 was confirmed using western blotting of UV exposed skin equivalents and ex vivo skin protein extracts. In conclusion, our study indicates the potency of a proteomic approach to study UV-induced changes in a tissue culture system mimicking human skin as well as excised human skin. PMID- 15816842 TI - Ultraviolet A irradiation induces NF-E2-related factor 2 activation in dermal fibroblasts: protective role in UVA-induced apoptosis. AB - Ultraviolet (UV) radiation is one of the most important environmental factors involved in the pathogenesis of skin aging and cancer. Many harmful effects of UV radiation are associated with the generation of reactive oxygen species, and cellular antioxidants act to prevent the occurrence and reduce the severity of UV induced skin disorders. Transcription factor NF-E2-related Factor 2 (Nrf2) and its cytoplasmic anchor protein Kelch-like-ECH-associated protein 1 (Keap1) are central regulators of the cellular antioxidant response. In this study, we investigated the effects of UV irradiation on the activation of Nrf2 in dermal fibroblasts. We found that UVA irradiation, but not UVB, causes nuclear translocation and accumulation of Nrf2 by a factor of 6.5 as compared with unirradiated controls. The nuclear accumulation of Nrf2 induced by UVA was enhanced by the photosensitizer hematoporphyrin. To evaluate the protective role of Nrf2 against UVA radiation, we examined UVA-induced apoptosis using dermal fibroblasts derived from nrf2 or keap1 gene knockout mice. Whereas disruption of nrf2 increased the number of apoptotic cells following UVA irradiation by 1.7 fold, disruption of keap1 decreased the apoptotic cell number by half as compared with wild-type controls. These findings thus demonstrate that the Nrf2-Keap1 pathway plays an important role in the protection of the skin against UVA irradiation. PMID- 15816843 TI - T-cadherin negatively regulates the proliferation of cutaneous squamous carcinoma cells. AB - T-cadherin is a unique member of the cadherin superfamily that lacks the transmembrane and cytoplasmic domains, and is instead linked to the cell membrane via a glycosyl-phosphatidylinositol anchor. We previously reported that T cadherin was specifically expressed on the basal keratinocytes of the epidermis, and the expression of T-cadherin was significantly reduced in invasive cutaneous squamous cell carcinoma (SCC) and in the lesional skin of psoriasis vulgaris. In this study, to obtain an insight into the role of T-cadherin in keratinocytes, we used transfection methods and examined the effect of overexpression or knockdown of T-cadherin in immortalized keratinocyte cell lines derived from SCC. T cadherin overexpressed cells showed clearly reduced cell proliferation, but the influence of cell-cell adhesiveness and cell mobility was not detected. Using a tetracycline-regulated expression system, we also confirmed that the suppression of cell proliferation was dependent on the expression level of T-cadherin. Cell cycle analysis demonstrated that over expression of T-cadherin induced a delay in the G(2)/M phase. Our findings suggest that T-cadherin acts as an endogenous negative regulator of keratinocyte proliferation and its inactivation is the cause for keratinocyte hyperproliferation in SCC or in psoriasis vulgaris. PMID- 15816844 TI - Gangliosides inhibit urokinase-type plasminogen activator (uPA)-dependent squamous carcinoma cell migration by preventing uPA receptor/alphabeta integrin/epidermal growth factor receptor interactions. AB - The interaction of the urokinase-type plasminogen activator (uPA) receptor (uPAR) with integrins plays a critical role in the regulation of cell adhesion and migration. However, the molecular events underlying the modulation of the interaction of uPAR and integrin are poorly understood. Gangliosides are thought to regulate epithelial cell adhesion and migration by inhibiting alpha(5)beta(1) integrin and epidermal growth factor receptor (EGFR) signaling. We report here that increases in the expression of ganglioside NeuAcalpha2-->3Galbeta1- >3GalNAcbeta1-->4(NeuAcalpha2-->8NeuAcalpha2-->3)Galbeta1-->4Glcbeta1-Cer (GT1b) or NeuAcalpha2-->3Galbeta1-->4Glcbeta1-Cer (GM3) inhibit uPA-dependent cell migration by preventing the association of uPAR with alpha(5)beta(1) integrin or uPAR/alpha(5)beta(1) integrin with the EGFR, respectively. As a result, uPA dependent focal adhesion kinase (FAK) and integrin-mediated EGFR signaling are suppressed. Both gangliosides inhibit uPAR signaling-stimulated migration; however, GM3 inhibits uPA-induced EGFR phosphorylation by blocking the crosstalk between integrin and EGFR, whereas GT1b suppresses both uPA-induced FAK and EGFR activation by preventing the activation of integrin alpha(5)beta(1). PMID- 15816845 TI - Matrilysin-2 (matrix metalloproteinase-26) is upregulated in keratinocytes during wound repair and early skin carcinogenesis. AB - Matrilysin-2 (matrix metalloproteinase (MMP)-26) is a small protein of the MMP family expressed in some epithelial carcinomas and normal tissues. We studied its role in benign skin disorders characterized by epithelial proliferation, in wound repair, skin cancer, and regulation in keratinocyte (KC) cultures. MMP-26 is expressed by laminin-5-positive KC in the migrating area during wound repair, in benign skin disorders characterized by inflammation and microdisruptions of basement membrane, but in intact skin only in hair follicles. It was detected in occasional atypical KC in pre-malignant lesions but not in basal cell cancer islands. Although MMP-26 was expressed in grades I and II squamous cell cancers (SCC), it was not present in dedifferentiated grade III tumors. MMP-26 was neither co-expressed with its close homologue matrilysin-1 nor with the proliferation marker Ki-67. But in tissue samples it either co-localized or was detected in adjacent cells of same regions with the tumor suppressor p16. In KC and HaCaT cell cultures, 12-phorbol-13-myristate-acetate, epidermal growth factor, tumor necrosis factor-alpha, transforming growth factor-beta1, interleukin-1 (IL-1)beta, IL-6, insulin-like growth factor, gamma-IFN, retinoic acid, dexamethasone, four matrices or ras-transformation were unable to upregulate MMP-26 expression. The expression pattern of MMP-26 suggests that it may be upregulated in basal KC even without tumorigenesis because of altered cell matrix interactions and inflammation and, unlike most MMP, becomes downregulated during histological dedifferentiation of SCC. Thus, lack of MMP-26 in SCC could be a marker of aggressive growth. PMID- 15816846 TI - Molecular crosstalk of the ultraviolet a and ultraviolet B signaling responses at the level of mitogen-activated protein kinases. PMID- 15816847 TI - Autofluorescence in the stem cell region of the hair follicle bulge. PMID- 15816848 TI - Genotype-phenotype correlation in recessive dystrophic epidermolysis bullosa: when missense doesn't make sense. PMID- 15816849 TI - Thromboxane A receptor-mediated cell proliferation, survival and gene expression in oligodendrocytes. AB - Thromboxane A(2) receptors (TP) were previously localized to discrete regions in the rat brain on myelinated fiber tracts and oligodendrocytes (OLGs). The present studies extended these findings and investigated the effects of TP signaling on cell proliferation, survival, and gene expression in OLG progenitor cells (OPCs) and OLGs. It was found that the TP agonist, U46619 stimulated the proliferation of OPCs and promoted the survival of mature OLGs. Examination of the early gene expression events involved in OPC proliferation, revealed that c-fos expression was substantially increased by U46619 stimulation. Treatment of OPCs or OLGs with U46619 caused activation of the mitogen-activated protein kinases (MAPK) ERK 1/2. In OPCs this activation was blocked by inhibition of src. However, in OLGs this phosphorylation was not only blocked by inhibition of src but also by inhibition of protein kinase C (PKC). Furthermore, U46619 was found to increase CREB phosphorylation in both OPCs and OLGs. Similar to ERK 1/2 activation, there was a divergence in the mechanism of the TP-mediated CREB response for each cell type. Specifically, U46619 activation was attenuated by src and protein kinase A (PKA) inhibition in OPCs, whereas in OLGs this effect was blocked by inhibition of src, PKA as well as by inhibition of PKC. Collectively, these results provide the first demonstration that TP-activated nuclear signaling events are involved in the proliferation of OPCs, the survival of mature OLGs, and the stimulation of gene expression. PMID- 15816850 TI - Sequestration of serum response factor in the hippocampus impairs long-term spatial memory. AB - The formation of long-term memory has been shown to require protein kinase mediated gene expression. One such kinase, mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK), can lead to the phosphorylation of serum response factor (SRF) and Elk-1, enhancing the expression of target genes. However, a direct involvement of these transcription factors in memory storage has not been demonstrated. We have employed an oligonucleotide decoy technique to interrogate SRF and Elk-1. Previously, it has been shown that intra-amygdalal infusion of small double-stranded decoy oligonucleotides for nuclear factor-kappaB (NFkappaB) can impair long-term memory for fear-potentiated startle. Using this approach, we found that intra hippocampal infusion of NFkappaB decoy oligonucleotides also impairs long-term spatial memory, consistent with a role for this factor in long-term memory storage. Decoy oligonucleotides containing the binding site for SRF, as confirmed by shift-western, did not influence memory acquisition but impaired long-term spatial memory. Analysis of search behavior during the transfer test revealed deficits consistent with a loss of precise platform location information. In contrast, oligonucleotides with a binding site for either Elk-1 or another target of ERK activity, SMAD3/SMAD4, did not interfere with memory formation or storage. These findings suggest that SRF-mediated gene expression is required for long term spatial memory. PMID- 15816851 TI - Puromycin-based purification of rat brain capillary endothelial cell cultures. Effect on the expression of blood-brain barrier-specific properties. AB - One of the main difficulties with primary rat brain endothelial cell (RBEC) cultures is obtaining pure cultures. The variation in purity limits the achievement of in vitro models of the rat blood-brain barrier. As P-glycoprotein expression is known to be much higher in RBECs than in any contaminating cells, we have tested the effect of five P-glycoprotein substrates (vincristine, vinblastine, colchicine, puromycin and doxorubicin) on RBEC cultures, assuming that RBECs would resist the treatment with these toxic compounds whereas contaminating cells would not. Treatment with either 4 microg/mL puromycin for the first 2 days of culture or 3 microg/mL puromycin for the first 3 days showed the best results without causing toxicity to the cells. Transendothelial electrical resistance was significantly increased in cell monolayers treated with puromycin compared with untreated cell monolayers. When cocultured with astrocytes in the presence of cAMP, the puromycin-treated RBEC monolayer showed a highly reduced permeability to sodium fluorescein (down to 0.75 x 10(-6) cm/s) and a high electrical resistance (up to 500 Omega x cm(2)). In conclusion, this method of RBEC purification will allow the production of in vitro models of the rat blood-brain barrier for cellular and molecular biology studies as well as pharmacological investigations. PMID- 15816852 TI - Activated mitogen-activated protein kinase kinase 7 redistributes to the cytosol and binds to Jun N-terminal kinase-interacting protein 1 involving oxidative stress during early reperfusion in rat hippocampal CA1 region. AB - Mitogen-activated protein kinase kinase (MKK) 7, a specific upstream activator of Jun N-terminal kinases (JNKs) in the stress-activated protein kinase (SAPK)/JNK signaling pathway, plays an important role in response to global cerebral ischemia. We investigated the subcellular localization of activated (phosphorylated) MKK (p-MKK) 7 using western blotting, immunoprecipitation and immunohistochemistry analysis in rat hippocampus. Transient forebrain ischemia was induced by the four-vessel occlusion method on Sprague-Dawley rats. Our results showed that both protein expression and activation of MKK7 were increased rapidly with peaks at 10 min of reperfusion in the nucleus of the hippocampal CA1 region. Simultaneously, in the cytosol activated MKK7 enhanced gradually and peaked at 30 min of reperfusion. In addition, we also detected JNK-interacting protein (JIP) 1, which accumulated in the perinuclear region of neurons at 30 min of reperfusion. Interestingly, at the same time-point the binding of JIP-1 to p MKK7 reached a maximum. Consequently, we concluded that MKK7 was rapidly activated and then translocated from the nucleus to the cytosol depending on its activation in the hippocampal CA1 region. To further elucidate the possible mechanism of MKK7 activation and translocation, the antioxidant N-acetylcysteine was injected into the rats 20 min before ischemia. The result showed that the levels of MKK7 activation, translocation and binding of p-MKK7 to JIP-1 were obviously limited by N-acetylcysteine in the cytosol at 30 min after reperfusion. The findings suggested that MKK7 activation, translocation and binding to JIP-1 were closely associated with reactive oxygen species and might play a pivotal role in the activation of the JNK signaling pathway in brain ischemic injury. PMID- 15816853 TI - Endocannabinoid metabolism in human glioblastomas and meningiomas compared to human non-tumour brain tissue. AB - The endogenous levels of the two cannabinoid receptor ligands 2-arachidonoyl glycerol and anandamide, and their respective congeners, monoacyl glycerols and N acylethanolamines, as well as the phospholipid precursors of N-acylethanolamines, were measured by gas chromatography-mass spectrometry in glioblastoma (WHO grade IV) tissue and meningioma (WHO grade I) tissue and compared with human non-tumour brain tissue. Furthermore, the metabolic turnover of N-acylethanolamines was compared by measurements of the enzymatic activity of N-acyltransferase, N acylphosphatidylethanolamine-hydrolysing phospholipase D and fatty acid amide hydrolase in the same three types of tissue. Glioblastomas were characterized by enhanced levels of N-acylethanolamines (eightfold, 128 +/- 59 pmol/micromol lipid phosphorus) including anandamide (17-fold, 4.6 +/- 3.1 pmol/micromol lipid phosphorus) and several species of N-acylphosphatidylethanolamines (three to eightfold). This was accompanied by a more than 60% reduction in the enzyme activities of N-acylphosphatidylethanolamine-hydrolysing phospholipase D and fatty acid amide hydrolase. By contrast, meningiomas were characterized by a massively enhanced level of 2-monoacyl glycerols (20-fold, 2293 +/- 361 pmol/micromol lipid phosphorus) including 2-arachidonoyl glycerol (20-fold, 1524 +/- 361 pmol/micromol lipid phosphorus). This was accompanied by an enhanced in vitro conversion of phosphatidylcholine to monoacyl glycerol (fivefold). The enhanced level of the 2-arachidonoyl glycerol, anandamide and other N acylethanolamines detected in the two types of tumour tissue may possibly act as endogenous anti-tumour mediators by stimulation of both cannabinoid and non cannabinoid receptor-mediated mechanisms. PMID- 15816854 TI - CGS21680 attenuates symptoms of Huntington's disease in a transgenic mouse model. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by a CAG trinucleotide expansion in exon 1 of the Huntingtin (Htt) gene. We show herein that in an HD transgenic mouse model (R6/2), daily administration of CGS21680 (CGS), an A(2A) adenosine receptor (A(2A)-R)-selective agonist, delayed the progressive deterioration of motor performance and prevented a reduction in brain weight. 3D-microMRI analysis revealed that CGS reversed the enlarged ventricle-to-brain ratio of R6/2 mice, with particular improvements in the left and right ventricles. (1)H-MRS showed that CGS significantly reduced the increased choline levels in the striatum. Immunohistochemical analyses further demonstrated that CGS reduced the size of ubiquitin-positive neuronal intranuclear inclusions (NIIs) in the striatum of R6/2 mice and ameliorated mutant Htt aggregation in a striatal progenitor cell line overexpressing mutant Htt with expanded polyQ. Moreover, chronic CGS treatment normalized the elevated blood glucose levels and reduced the overactivation of a major metabolic sensor [5'AMP-activated protein kinase (AMPK)] in the striatum of R6/2 mice. Since AMPK is a master switch for energy metabolism, modulation of energy dysfunction caused by the mutant Htt might contribute to the beneficial effects of CGS. Collectively, CGS is a potential drug candidate for the treatment of HD. PMID- 15816855 TI - Differential phosphodiesterase expression and cytosolic Ca2+ in human CNS tumour cells and in non-malignant and malignant cells of rat origin. AB - A promising attempt in the field of tumour therapy is the modulation of intracellular, proliferation-associated signalling pathways. The role of cyclic nucleotide phosphodiesterases (PDEs), key enzymes in cAMP/cGMP signal transduction, was investigated in two human CNS tumour cell lines as well as in the rat glioblastoma cell line C6 in comparison with rat cerebellar astrocytes with the emphasis on target evaluation. We found differential PDE expression patterns in human CNS tumour cell lines as well as in CNS cells of rat origin. In human glioblastoma cells, intracellular cAMP and Ca(2+) levels correlated well with the PDE expression pattern. There were, however, marked differences in PDE expression and Ca(2+) kinetics between the human glioblastoma cell lines. In contrast to human epithelial tumour cells, shown earlier by us to express significantly enhanced cAMP-specific PDE activity, this was not the case in rat glioblastoma cells compared with non-malignant rat astrocytes. Despite different levels of PDE1 and PDE4 expression and activity, cyclic nucleotide and Ca(2+) levels in non-malignant and malignant rat CNS cells were similar. These in vitro data do not support the concept of PDE1C representing a target exploitable for drug treatment of malignant CNS tumours. PMID- 15816856 TI - Expression of human FE65 in amyloid precursor protein transgenic mice is associated with a reduction in beta-amyloid load. AB - FE65 is an adaptor protein that interacts with the cytoplasmic tail of the amyloid precursor protein (APP). In cultured non-neuronal cells, the formation of the FE65-APP complex is a key element for the modulation of APP processing, signalling and beta-amyloid (Abeta) production. The functions of FE65 in vivo, including its role in the metabolism of neuronal APP, remain to be investigated. In this study, transgenic mice expressing human FE65 were generated and crossbred with APP transgenic mice, known to develop Abeta deposits at 6 months of age. Compared with APP mice, APP/FE65 double transgenic mice exhibited a lower Abeta accumulation in the cerebral cortex as demonstrated by immunohistochemistry and immunoassay, and a lower level of APP-CTFs. The reduced accumulation of Abeta in APP/FE65 double transgenics, compared with APP mice, could be linked to the low Abeta42 level observed at 4 months of age and to the lower APP-CTFs levels. The present work provides evidence that FE65 plays a role in the regulation of APP processing in an in vivo model. PMID- 15816857 TI - Retinoic acid activates human secretin gene expression by Sp proteins and nuclear factor I in neuronal SH-SY5Y cells. AB - Secretin is a neuropeptide that is expressed in distinct central neurones. As there is no information on how the secretin gene is regulated in neuronal cells, a well established neuronal differentiation cell model, SH-SY5Y, was used to study transcriptional regulation of the human secretin gene. High secretin transcript and peptide levels were found in this cell, and secretin gene expression and promoter activity were up-regulated upon all-trans retinoic acid (RA) treatment. Within the promoter, a functional GC-box 1 (-131 from ATG, relative to the ATG initiation codon) was found to be regulated by a brain specific Sp protein, Sp4, and ubiquitous factors Sp1 and Sp3. The human secretin gene in SH-SY5Y cells is controlled by the (Sp1 + Sp4)/Sp3 ratio and the RA induced activation is a partial result of a decrease in Sp3 levels. In addition to the GC-box 1, an N1 motif in close proximity was also responsible for RA induced secretin gene activation. Competitive gel mobility shift and southwestern blot studies revealed binding of Nuclear Factor I (NFI) with the N1 motif. Overexpression of NFI-C increased promoter activity upon RA treatment. Consistent with this observation, NFI-C transcript levels were augmented after RA treatment. We conclude that RA induction of the secretin gene in neuronal cells is regulated by the combined actions of reducing Sp3 and increasing NFI-C expression. PMID- 15816858 TI - Overexpression of human copper/zinc-superoxide dismutase in transgenic animals attenuates the reduction of apurinic/apyrimidinic endonuclease expression in neurons after in vitro ischemia and after transient global cerebral ischemia. AB - Oxidative stress after ischemia/reperfusion has been shown to induce DNA damage and subsequent DNA repair activity. Apurinic/apyrimidinic endonuclease (APE) is a multifunctional protein in the DNA base excision repair pathway which repairs apurinic/apyrimidinic sites in DNA. We investigated the involvement of oxidative stress and expression of APE in neurons after oxygen-glucose deprivation and after global cerebral ischemia. Our results suggest that overexpression of human copper/zinc-superoxide dismutase reduced oxidative stress with a subsequent decrease in APE expression. Production of oxygen free radicals and inhibition of the base excision repair pathway may play pivotal roles in the cell death pathway after ischemia. PMID- 15816859 TI - Alcohol-responsive genes in the frontal cortex and nucleus accumbens of human alcoholics. AB - The molecular processes underlying alcohol dependence are not fully understood. Many characteristic behaviours result from neuroadaptations in the mesocorticolimbic system. In addition, alcoholism is associated with a distinct neuropathology. To elucidate the molecular basis of these features, we compared the RNA expression profile of the nucleus accumbens and prefrontal cortex of human brain from matched individual alcoholic and control cases using cDNA microarrays. Approximately 6% of genes with a marked alcohol response were common to the two brain regions. Alcohol-responsive genes were grouped into 11 functional categories. Predominant alcohol-responsive genes in the prefrontal cortex were those encoding DNA-binding proteins including transcription factors and repair proteins. There was also a down-regulation of genes encoding mitochondrial proteins, which could result in disrupted mitochondrial function and energy production leading to oxidative stress. Other alcohol-responsive genes in the prefrontal cortex were associated with neuroprotection/apoptosis. In contrast, in the nucleus accumbens, alcohol-responsive genes were associated with vesicle formation and regulation of cell architecture, which suggests a neuroadaptation to chronic alcohol exposure at the level of synaptic structure and function. Our data are in keeping with the previously reported alcoholism related pathology characteristic of the prefrontal cortex, but suggest a persistent decrease in neurotransmission and changes in plasticity in the nucleus accumbens of the alcoholic. PMID- 15816860 TI - Selective serotonin reuptake blockade increases extracellular dopamine in noradrenaline-rich isocortical but not prefrontal areas: dependence on serotonin 1A receptors and independence from noradrenergic innervation. AB - The present study investigated the effects of two serotonin (5-HT) uptake inhibitors, citalopram and paroxetine, and of a non-selective noradrenaline (NA) and 5-HT uptake blocker, imipramine, on extracellular NA and dopamine (DA) in the prefrontal cortex (PfCX), parietal cortex (ParCX) and occipital cortex (OccCX). Citalopram, the most selective 5-HT uptake blocker, increased dialysate DA in the OccCX and ParCX but not in the PfCX and this effect was prevented in the OccCX by WAY-100635, an antagonist of serotonin-1A (5-HT(1A)) receptors, but not by dorsal noradrenergic bundle (DNAB) lesions that reduced to unmeasurable levels basal dialysate NA but did not affect dialysate DA. Paroxetine, a less selective 5-HT uptake inhibitor than citalopram, at the dose of 5 mg/kg, increased DA in the OccCX but not in the PfCX; however, at doses of 10 mg/kg, which increase PfCX NA, paroxetine increased DA also in this area. Imipramine increased dialysate DA and NA both in the PfCX and in the OccCX and this effect was abolished by DNAB lesions and was reduced but not abolished by WAY-100635. Administration of doses of reboxetine and citalopram that do not increase DA release in the OccCX if given separately, markedly increased DA when combined. These results indicate that endogenous 5-HT, raised by selective blockade of the 5-HT carrier, can increase extracellular DA in the OccCX and in the ParCX by stimulating 5-HT(1A) receptors independently from the presence of NA terminals, although blockade of 5 HT and NA carrier can strongly interact to raise extracellular DA in this area. These observations are consistent with the existence of DA neurons separate from the NA ones contributing to extracellular DA even in NA-rich/DA poor isocortical areas. PMID- 15816861 TI - Cloning and characterization of a family C orphan G-protein coupled receptor. AB - Members of the family C receptors within the G-protein coupled receptor superfamily include the metabotropic glutamate receptors, GABA(B) receptors, the calcium-sensing receptor (CaSR), the V2R pheromone receptors, the T1R taste receptors, and a small group of uncharacterized orphan receptors. We have cloned and studied the mouse GPRC6A family C orphan receptor. The open reading frame codes for a protein with highest sequence identity to the fish 5.24 odorant receptor and the mammalian CaSR. The gene structure shows a striking resemblance to that of the CaSR. Results from RT-PCR analyses showed that mouse GPRC6A mRNA is expressed in mouse brain, skeletal muscle, heart, lung, spleen, kidney, liver, and in the early stage mouse embryo. Immunocytochemical analysis of the cloned mouse GPRC6A cDNA expressed in human embryonic kidney 293 cells demonstrated that GPRC6A was present on the plasma membrane, as well as in the endoplasmic reticulum and nuclear envelope membranes of transfected cells. A chimeric cDNA construct in which the extracellular ligand binding domain of the fish 5.24 amino acid-activated odorant receptor was ligated to the complementary downstream sequence of the mouse GPRC6A receptor indicated that GPRC6A is coupled to phosphoinositol turnover and release of intracellular calcium. Further studies with mouse GPRC6A expressed in Xenopus laevis oocytes demonstrated that this receptor possesses a pharmacological profile resembling that of the fish 5.24 odorant receptor. These findings suggest that GPRC6A may function as the receptor component of a novel cellular transmitter system in mammals. PMID- 15816862 TI - Astroglial cytoprotection by erythropoietin pre-conditioning: implications for ischemic and degenerative CNS disorders. AB - Erythropoietin (Epo) is a glycoprotein secreted by the kidney in response to hypoxia that stimulates erythropoiesis through interaction with cell surface Epo receptors. Pre-treatment with Epo has been shown to protect neurons in models of ischemic injury. The mechanism responsible for this neuroprotection and the effects of Epo on astroglial and other non-neuronal cell populations remain unknown. In the present study, we determined whether Epo pre-treatment protects neonatal rat astrocytes from apoptotic cell death resulting from treatment with nitric oxide, staurosporine (STS) and arsenic trioxide and possible mechanisms mediating Epo-related cytoprotection. Epo (5-20 U/mL) significantly attenuated multiple hallmarks of apoptotic cell death in astroglia exposed to nitric oxide and STS but not arsenic trioxide. Epo (20 U/mL) induced mild oxidative stress as shown by increases in heme oxygenase (HO)-1 mRNA and protein expression that could be suppressed by antioxidant coadministration. Moreover, coincubation with tin-mesoporphyrin, a competitive inhibitor of HO activity, abrogated the cytoprotective effects of Epo (20 U/mL) in the face of STS treatment. Thus, induction of the ho-1 gene may contribute to the glioprotection accruing from high-dose Epo exposure. Epo may augment astroglial resistance to certain chemical stressors by oxidative stress-dependent and -independent mechanisms. PMID- 15816863 TI - Integrative role of cPLA with COX-2 and the effect of non-steriodal anti inflammatory drugs in a transgenic mouse model of amyotrophic lateral sclerosis. AB - Cyclooxygenase-2 (COX-2) is a key molecule in the inflammatory pathway in amyotrophic lateral sclerosis (ALS). Cytosolic phospholipase A (cPLA2) is an important enzyme providing substrate for cyclooxygenases. We therefore examined cPLA2 expression in human ALS and mutant Cu/Zn superoxide dismutase (SOD1) transgenic mice and its relation to COX-2. Immunohistochemistry and real-time RT PCR revealed elevated cPLA2 protein and its mRNA levels in the lumbar spinal cord of mutant SOD1 mice. COX-2 immunoreactivity was increased in lumbar spinal cord sections from both familial ALS (FALS) and sporadic ALS (SALS) as compared to controls, and cPLA2 immunoreactivity was increased in a patient with FALS. Oral administration of the non-selective cyclooxygenase (COX) inhibitor, sulindac, extended the survival (by 10%) of G93A SOD1 mice as compared to littermate controls. Sulindac, as well as the selective COX-2 inhibitors, rofecoxib and celecoxib reduced cPLA2 immunoreactivity in the lumbar spinal cord of G93A transgenic mice. Sulindac treatment preserved motor neurons, and reduced microglial activation and astrocytosis, in the spinal cord of G93A SOD1 transgenic mice. These results suggest that cPLA2 plays an important role in supplying arachidonic acid to the COX-2 driven inflammatory pathway in ALS associated with SOD1 mutations. PMID- 15816864 TI - Erythropoietin protects primary hippocampal neurons increasing the expression of brain-derived neurotrophic factor. AB - Erythropoietin, the principal regulator of erythroids progenitor cells, also promotes neuronal survival. Using primary cultures of rat hippocampal neurons, we investigated whether erythropoietin could mediate neuroprotection favouring the transcription of brain-derived neurotrophic factor (BDNF). Erythropoietin 2.7 nm reduced by approximately 50% the neuronal death triggered by the prototypic neurotoxicant trimethyltin (TMT) and time-dependently induced BDNF mRNA. This effect resulted in an increased production of biologically active BDNF, which led to a sustained activation of the specific BDNF receptor tyrosine kinase B (TrkB). Reduction of TMT-induced neuronal death by erythropoietin was specifically prevented by a neutralizing anti-BDNF antibody (15 microg/mL), indicating the involvement of this neurotrophin in erythropoietin neuroprotective effect. Intracerebroventricular administration of erythropoietin in mice significantly increases BDNF mRNA expression in brain, supporting the idea of the involvement of this neurotrophin in erythropoietin action within the CNS. BDNF expression in neuronal cells is induced by activation of voltage Ca(2+)-channels and recruitment of Ca(2+)-sensitive transcription factors. Consistently, 2.7 nm erythropoietin increased intracellular Ca(2+) in 5 min and cAMP response element binding protein (CREB) phosphorylation at Ser 133 in 30 min. Both effects were abolished by 1 microm nitrendipine, a selective blocker of L-type voltage Ca(2+) channels. These data demonstrate that erythropoietin activates the CREB transcription pathway and increases BDNF expression and production, which contributes to erythropoietin mediated neuroprotection. PMID- 15816865 TI - Alterations in the solubility and intracellular localization of parkin by several familial Parkinson's disease-linked point mutations. AB - Mutations in the parkin gene, which encodes a ubiquitin ligase, are currently recognized as the main contributor to familial forms of Parkinson's disease (PD). A simple assumption about the effects of PD-linked mutations in parkin is that they impair or ablate the enzyme activity. However, a number of recent studies, including ours, have indicated that many disease-linked point mutants of parkin retain substantial catalytic activity. To understand how the plethora of mutations on parkin contribute to its dysfunction, we have conducted a systematic analysis of a significant number of parkin point mutants (22 in total), which represent the majority of parkin missense/nonsense mutations reported to date. We found that more than half of these mutations, including many located outside of the parkin RING fingers, produce alteration in the solubility of parkin which influences its detergent extraction property. This mutation-mediated alteration in parkin solubility is also associated with its propensity to form intracellular, aggresome-like, protein aggregates. However, they do not represent sites where parkin substrates become sequestered. As protein aggregation sequesters the functional forms away from their normal sites of action, our results suggest that alterations in parkin solubility and intracellular localization may underlie the molecular basis of the loss of function caused by several of its mutations. PMID- 15816866 TI - Stage- and site-specific DNA demethylation during neural cell development from embryonic stem cells. AB - Activation of the transcription factor STAT3 is important for astrocyte differentiation during neural development. Demethylation of the methyl-CpG dinucleotide in the STAT3 binding site in the promoter of the glial fibrillary acidic protein (GFAP) gene, a marker for astrocytes, was previously shown to be a crucial cue for neural progenitors to express this gene in response to astrogenic signals during brain development. In this study, we analyzed the methylation status of the STAT3 binding site in the GFAP gene promoter during neural cell development from mouse embryonic stem (ES) cells in vitro. The CpG dinucleotide in the STAT3 binding site in the GFAP gene promoter exhibited a high incidence of cytidine-methylation in undifferentiated pluripotent ES cells. The high incidence of methylation of this particular cytidine was maintained in ES cell-derived neuroectoderm-like cells, but it underwent demethylation when the neural lineage cells became competent to express GFAP in response to a STAT3 activation signal. In contrast, hypermethylation of the CpG site was maintained in non-neural cells generated from the same ES cells. Progressive demethylation of the STAT3 binding site in the GFAP gene promoter was also observed in primary embryonic neuroepithelial cells during in vitro culture, whereas non-neural cells maintained hypermethylation of this site even after culture. Taken together, these results demonstrate that the astrocyte gene-specific cytidine-demethylation is programmed when neural progenitors from pluripotent cells are committed to a neural lineage that is capable of producing astrocytes. PMID- 15816867 TI - A family of octopamine [corrected] receptors that specifically induce cyclic AMP production or Ca2+ release in Drosophila melanogaster. AB - In invertebrates, the biogenic-amine octopamine is an important physiological regulator. It controls and modulates neuronal development, circadian rhythm, locomotion, 'fight or flight' responses, as well as learning and memory. Octopamine mediates its effects by activation of different GTP-binding protein (G protein)-coupled receptor types, which induce either cAMP production or Ca(2+) release. Here we describe the functional characterization of two genes from Drosophila melanogaster that encode three octopamine receptors. The first gene (Dmoa1) codes for two polypeptides that are generated by alternative splicing. When heterologously expressed, both receptors cause oscillatory increases of the intracellular Ca(2+) concentration in response to applying nanomolar concentrations of octopamine. The second gene (Dmoa2) codes for a receptor that specifically activates adenylate cyclase and causes a rise of intracellular cAMP with an EC(50) of approximately 3 x 10(-8) m octopamine. Tyramine, the precursor of octopamine biosynthesis, activates all three receptors at > or = 100-fold higher concentrations, whereas dopamine and serotonin are non-effective. Developmental expression of Dmoa genes was assessed by RT-PCR. Overlapping but not identical expression patterns were observed for the individual transcripts. The genes characterized in this report encode unique receptors that display signature properties of native octopamine receptors. PMID- 15816868 TI - Functional SDF1 alpha/CXCR4 signaling in the developing spinal cord. AB - Stromal cell-derived factor (SDF1) and its cognate receptor CXCR4 have been shown to play a central role in the development of the cerebellum, hippocampus, and neocortex. However, little is known about the functions of SDF1/CXCR4 in early spinal cord progenitor cell differentiation. Here, we show that a functional SDF1alpha/CXCR4 signaling pathway is present in developing spinal cord cells (a spliced variant of SDF1). RT-PCR analysis of SDF1alpha and CXCR4 showed that they were present in E10.5 neural tube and their expression increased as neuroepithelial cells differentiated into more committed spinal cord progenitors. Stimulation of the more differentiated progenitors (E14.5) with SDF1alpha resulted in rapid activation of the extracellular signal-regulated kinase (ERK)1/2. This SDF1alpha-induced ERK activity was dose dependent and could be inhibited by pre-treatment of the cells with either pertussis toxin, an inactivator of G-protein-coupled receptors, or PD98059, a MEK1 inhibitor. Concomitant with ERK activation, SDF1alpha also activated the downstream transcription factor Ets, a substrate for ERK phosphorylation. Further, downstream activation of genes associated with cell survival, differentiation and migration was assessed using a G-protein-coupled receptor pathway-focused microarray. We found that 23 genes, including PDK1, Egr-1, Grm5, and E-selectin, were up-regulated by SDF1alpha. Furthermore, SDF1alpha induced chemotaxis in both neural and glial progenitors in in vitro migration assays. Pre-treatment of the cells with either pertussis toxin or PD98059 completely inhibited SDF1alpha induced chemotaxis. Thus, our data suggest that SDF1alpha may function through a CXCR4/ERK/Ets-linked signalling pathway in spinal cord neural development to modulate migration of progenitor cells. PMID- 15816869 TI - Impaired long-term potentiation in c-Jun N-terminal kinase 2-deficient mice. AB - c-Jun N-terminal kinases (JNKs) are thought to be involved in regulating synaptic plasticity. We therefore investigated the specific role of JNK2 in modulating long-term potentiation (LTP) in hippocampus during development, using JNK2 deficient mice. The morphological structure and the numbers of both NeuN, a specific neuronal marker, and GABA-positive neurons in the hippocampal areas were similar in wild-type and Jnk2(-/-) mice. Western blot analysis revealed that JNK2 expression was higher and stable at 1 and 3 months of age, but JNK1 levels were lower at 1 month of age and almost undetectable in 3-month-old wild-type mice. In contrast to wild-type mice, there was a significant increase in JNK1 expression in JNK2 mutant mice, especially at 1 month of age. Electrophysiological studies demonstrated that LTP was impaired in both the CA1 and CA3 regions in 1-month old, but not in adult, Jnk2(-/-) mice, probably owing to decreased presynaptic neurotransmitter release. Moreover, late-phase LTP, but not early-phase LTP, was impaired in the Jnk2(-/-) adult mice, suggesting that JNK2 plays a role in transforming early LTP to late LTP. Together, the data highlight the specific role of JNK2 in hippocampal synaptic plasticity during development. PMID- 15816870 TI - Sp1 and Sp3 activate transcription of the human dopamine transporter gene. AB - The dopamine transporter is a plasma membrane protein that controls extracellular concentrations of the neurotransmitter dopamine. The physiological importance of the DAT provides the impetus for studies aimed at understanding the molecular mechanisms underlying regulation of the DAT gene. In this study, we identified a DAT-expressing neuroblastoma cell line (SK-N-AS) and employed it to investigate the transcriptional regulation of the human DAT gene. Two GC boxes (located at 130 and -60, respectively, relative to the transcriptional start site) were identified as important cis-acting elements mediating DAT promoter activity in dopaminergic SK-N-AS cells. Utilizing Sp-deficient Drosophila Schneider line (SL 2) cells, we showed that both Sp1 and Sp3 are strong activators of DAT transcriptional activity. Differential binding of Sp1 and Sp3 to the two GC boxes was demonstrated by electrophoretic mobility shift assays and super-shift assays. Our results indicate that the Sp1 family of proteins plays an important role in controlling the expression of the dopamine transporter gene within dopaminergic neurons. PMID- 15816871 TI - Histone deacetylase inhibitors decrease reelin promoter methylation in vitro. AB - We investigated the effects of agents that induce reelin mRNA expression in vitro on the methylation status of the human reelin promoter in neural progenitor cells (NT2). NT2 cells were treated with the histone deacetylase inhibitors, trichostatin A (TSA) and valproic acid (VPA), and the methylation inhibitor aza 2'-deoxycytidine (AZA) for various times. All three drugs reduced the methylation profile of the reelin promoter relative to untreated cells. The acetylation status of histones H3 and H4 increased following treatment with VPA and TSA at times as short as 15 min following treatment; a result consistent with the reported mode of action of these drugs. Chromatin immunoprecipitation experiments showed that these changes were accompanied by changes occurring at the level of the reelin promoter as well. Interestingly, AZA decreased reelin promoter methylation without concomittantly increasing histone acetylation. In fact, after prolonged treatments with AZA, the acetylation status of histones H3 and H4 decreased relative to untreated cells. We also observed a trend towards reduced methylated H3 after 18 h treatment with TSA and VPA. Our data indicate that while TSA and VPA act to increase histone acetylation and reduce promoter methylation, AZA acts only to decrease the amount of reelin promoter methylation. PMID- 15816872 TI - Uncoupling protein 2 protects dopaminergic neurons from acute 1,2,3,6-methyl phenyl-tetrahydropyridine toxicity. AB - Oxidative stress is implicated in the death of dopaminergic neurons in sporadic forms of Parkinson's disease. Because oxidative stress can be modulated endogenously by uncoupling proteins (UCPs), we hypothesized that specific neuronal expression of UCP2, one member of the UCP family that is rapidly induced in the CNS following insults, could confer neuroprotection in a mouse model of Parkinson's disease. We generated transgenic mice overexpressing UCP2 in catecholaminergic neurons under the control of the tyrosine hydroxylase promoter (TH-UCP2). In these mice, dopaminergic neurons of the substantia nigra showed a twofold elevation in UCP2 expression, elevated uncoupling of their mitochondria, and a marked reduction in indicators of oxidative stress, an effect also observed in the striatum. Upon acute exposure to 1,2,3,6-methyl-phenyl-tetrahydropyridine, TH-UCP2 mice showed neuroprotection and retention of locomotor functions. Our data suggest that UCP2 may represent a drug target for slowing the progression of Parkinson's disease. PMID- 15816873 TI - Control of cyclin-dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability. AB - Although the roles of cyclin-dependent kinase 5 (Cdk5) in neurodevelopment and neurodegeneration have been studied extensively, regulation of Cdk5 activity has remained largely unexplored. We report here that glutamate, acting via NMDA or kainate receptors, can induce a transient Ca(2+)/calmodulin-dependent activation of Cdk5 that results in enhanced autophosphorylation and proteasome-dependent degradation of a Cdk5 activator p35, and thus ultimately down-regulation of Cdk5 activity. The relevance of this regulation to synaptic plasticity was examined in hippocampal slices using theta burst stimulation. p35(-/-) mice exhibited a lower threshold for induction of long-term potentiation. Thus excitatory glutamatergic neurotransmission regulates Cdk5 activity through p35 degradation, and this pathway may contribute to plasticity. PMID- 15816874 TI - Chronic renal failure after liver transplantation. PMID- 15816875 TI - Innate immunity and organ transplantation: the potential role of toll-like receptors. AB - Traditionally, the recognition and tolerance of transplanted grafts has been considered to be within the realm of the adaptive immune system. Innate immunity, on the other hand, as the first line of host defense, plays a role in fighting against invading microorganisms. Recently, with the discovery of the Toll-like receptors (TLRs), the role of innate immune responses in the control of adaptive immunity has become a new area of interest. Emerging evidence suggests that in addition to responding to pathogen-associated molecular patterns of microorganisms, TLRs can be activated by endogenous ligands, expressed by mammalian cells. These 'danger signals' may participate in ischemia-reperfusion related organ damage and subsequently influence function and survival of transplanted grafts. Furthermore, it has been suggested that adaptive immune responses can enhance the acute inflammatory responses controlled by innate immunity in organ transplantation. This review addresses the potential involvement of TLRs in different stages of organ transplantation. Intriguing and controversial findings are presented and discussed in order to stimulate more attention to this emerging and potentially important area of research in organ transplantation. PMID- 15816876 TI - Paul E. Lacy, MD, PhD February 7, 1924 to February 15, 2005. PMID- 15816877 TI - CTLA4 engagement is required for induction of murine liver transplant spontaneous tolerance. AB - Liver transplantation in mice is accepted spontaneously in all strain combinations. The mechanisms remain largely undefined. We hypothesize that signaling via the B7-CTLA4 receptor pathway is required for induction of liver transplant tolerance. Liver transplantation was performed from B10 (H2(b)) to C3H (H2(k)) mice. The recipients received anti-mouse CTLA4 mAb 0.25 mg i.p. every other day post-operatively. Liver grafts in anti-CTLA4 mAb treated recipients were acutely rejected. The allo-specific proliferative responses, anti-donor CTL and NK cell activities of GIC and SC and the serum levels of IFN-gamma and IL-2 from anti-CTLA4 mAb treated recipients were elevated significantly in comparison to the control mice. The frequency of IFN-gamma and IL-2 producing cells were markedly increased also in the anti-CTLA4 treated recipients. The immunohistology of liver grafts from anti-CTLA4 mAb treated mice showed extensively increased lymphocyte infiltration in the portal and general parenchymal areas, and expanded T-cell area in the spleen, with a reduction in the frequency of apoptotic cells observed by TUNEL staining compared with control mice. Thus CTLA4 signaling is critical for murine liver transplant tolerance induction. CTLA4 blockade promotes donor specific T-cell activation, cytotoxicity and Th1 polarization; protects alloreactive T cells from apoptotic death and induces liver allograft acute rejection. PMID- 15816878 TI - Cyclosporine interacts with mycophenolic acid by inhibiting the multidrug resistance-associated protein 2. AB - In mycophenolate mofetil (MMF)-treated organ transplant recipients, lower mycophenolic acid (MPA) plasma concentrations have been found in cyclosporine (CsA) compared with tacrolimus (Tac)-based immunosuppressive regimens. We previously demonstrated that CsA decreases exposure to MPA and increases exposure to its metabolite MPA-glucuronide (MPAG), possibly by interfering with the biliary excretion of MPAG. To elucidate the role of the multidrug resistance associated protein (Mrp)-2 in the interaction between MMF and CsA, we treated three groups of 10 Mrp2-deficient rats (TR- rat) for 6 days with either vehicle, CsA (8 mg/kg) or Tac (4 mg/kg) by oral gavage. Hereafter, co-administration with MMF (20 mg/kg) was started in all groups and continued through day 14. The 24-h MPA/MPAG area under the concentration-time curve (AUC) was determined after single (day 7) and multiple MMF doses (day 14). On both study days, there were no significant differences in the mean MPA and MPAG AUC between CsA and Tac-treated animals. We conclude that the pharmacokinetics of MMF are comparable in Mrp2 deficient rats receiving either CsA or Tac as co-medication. This finding suggests that CsA-mediated inhibition of the biliary excretion of MPAG by the Mrp2 transporter is the mechanism responsible for the interaction between CsA and MMF. PMID- 15816879 TI - Lung preservation: pulmonary flush route affects bronchial mucosal temperature and expression of IFN-gamma and Gro in regional lymph nodes. AB - Optimal lung preservation via flush of the pulmonary vasculature minimizes early graft failure post-lung transplantation. We hypothesized that the route of pulmonary flush has differential effects on thermal gradients in the lung and expression of inflammatory mediators. Swine underwent antegrade flush (AG) via pulmonary artery; AG/RG: antegrade + retrograde flush via pulmonary veins or AG/BA: antegrade + bronchial artery flush via bronchial artery. Temperatures were recorded in bronchial mucosa and peribronchial lymph nodes. RT-PCR was utilized to detect cytokine gene expression in the nodes. AG/BA flush resulted in greatest cooling of bronchial mucosa and lymph nodes (p < 0.001). The route of flush did not affect expression of RANTES, MCP-1, IL-8, IL-1beta, TNF-alpha or IL-6. However, expression of Gro was reduced 4-h post-preservation in all groups. Only AG/BA resulted in decreased IFN-gamma transcripts. These data show that, compared to AG or AG/RG, AG/BA flush results in the greatest cooling of lung compartments and down regulates lymph node expression of a cytokine and chemokine that have key roles in inflammation and immunity. These data suggest that pulmonary flush via AG/BA during donor harvest may be optimal to decrease the risk of early graft failure. PMID- 15816880 TI - Inosculation of tissue-engineered capillaries with the host's vasculature in a reconstructed skin transplanted on mice. AB - The major limitation for the application of an autologous in vitro tissue engineered reconstructed skin (RS) for the treatment of burnt patients is the delayed vascularization of its relatively thick dermal avascular component, which may lead to graft necrosis. We have developed a human endothelialized reconstructed skin (ERS), combining keratinocytes, fibroblasts and endothelial cells (EC) in a collagen sponge. This skin substitute then spontaneously forms a network of capillary-like structures (CLS) in vitro. After transplantation to nude mice, we demonstrated that CLS containing mouse blood were observed underneath the epidermis in the ERS in less than 4 days, a delay comparable to our human skin control. In comparison, a 14-day period was necessary to achieve a similar result with the non-endothelialized RS. Furthermore, no mouse blood vessels were ever observed close to the epidermis before 14 days in the ERS and the RS. We thus concluded that the early vascularization observed in the ERS was most probably the result of inosculation of the CLS network with the host's capillaries, rather than neovascularization, which is a slower process. These results open exciting possibilities for the clinical application of many other tissue-engineered organs requiring a rapid vascularization. PMID- 15816881 TI - Warfarin or low-molecular-weight heparin therapy does not prolong pig-to-primate cardiac xenograft function. AB - Microvascular thrombosis is a prominent feature in cardiac delayed xenograft rejection (DXR). We investigated the impact of warfarin or low-molecular-weight heparin (LMWH) anti-coagulation on xenograft function using a heterotopic pig-to primate model. Donor hearts were from CD46 transgenic pigs and baboon immunosuppression included tacrolimus, sirolimus, anti-CD20 and TPC, an alpha galactosyl-polyethylene glycol conjugate. Three groups of animals were studied. Group 1 (n = 9) was treated with warfarin, Group 2 (n = 13) with LMWH and Group 3, received no anti-coagulant drugs. The median duration of xenograft function was 20 days (range 3-62 days), 18 days (range 5-109 days) and 15 days (range 4-53 days) in Groups 1 to 3 respectively. Anti-coagulation achieved the targeted international normalized prothrombin ratio (INR) and anti-factor Xa levels consistent with effective in vivo therapy yet, no significant impact on median xenograft function was observed. At rejection, a similar histology of thrombosis and ischemia was apparent in each group and the levels of fibrin deposition and platelet thrombi in rejected tissue was the same. Anti-coagulation with warfarin or LMWH did not have a significant impact on the onset of DXR and microvascular thrombosis. However, a role for specific anti-coagulant strategies to achieve long-term xenograft function cannot be excluded. PMID- 15816882 TI - Anti-CD154 mAb treatment but not recipient CD154 deficiency leads to long-term survival of xenogeneic islet grafts. AB - The aim of the study was to evaluate the role of recipient CD40 and CD154 in the rejection process of concordant and discordant islets xenotransplantation (Tx). Diabetic C57BL/6 mice, CD40- or CD154-knockout (KO) and complement C3-deficient (C3-/-) mice were transplanted with either rat or human islets. Group 1, C57BL/6 Tx without therapy; Group 2, C57BL/6 Tx with anti-CD154 monoclonal antibody (mAb) (MR1) therapy; Group 3, CD40-KO; and Group 4, CD154-KO Tx without therapy; Group 5, C3-/- Tx without therapy and Group 6, C3-/-Tx with MR1 therapy. Mixed lymphocyte reactions (MLR) were performed. Compared to Group 1, MR1 induced long term survival of xenografts in Group 2, but not in Group 6, survival of islets was not prolonged significantly in Groups 3 and 4. MLR responses in Group 2 were reduced approximately 50% compared to Group 1. In Groups 3, 4 and 6, MLR responses were not modified by the absence of CD40 or CD154 molecules, or MR1 and were similar to Group 1. Improved graft survival and reduced MLR responses in Group 2, but not in Group 6, could be explained by specific targeting of activated T cells with inactivation by complement- or cellular-mediated mechanisms. Rejection of xenografts and strong MLR responses in Groups 3 and 4 are possible through efficient activation of alternate pathways of costimulation. PMID- 15816883 TI - IDEC-131 (anti-CD154), sirolimus and donor-specific transfusion facilitate operational tolerance in non-human primates. AB - CD154-specific antibody therapy prevents allograft rejection in many experimental transplant models. However, initial clinical transplant trials with anti-CD154 have been disappointing suggesting the need for as of yet undetermined adjuvant therapy. In rodents, donor antigen (e.g., a donor blood transfusion), or mTOR inhibition (e.g., sirolimus), enhances anti-CD154's efficacy. We performed renal transplants in major histocompatibility complex-(MHC) mismatched rhesus monkeys and treated recipients with combinations of the CD154-specific antibody IDEC-131, and/or sirolimus, and/or a pre-transplant donor-specific transfusion (DST). Therapy was withdrawn after 3 months. Triple therapy prevented rejection during therapy in all animals and led to operational tolerance in three of five animals including donor-specific skin graft acceptance in the two animals tested. IDEC 131, sirolimus and DST are highly effective in preventing renal allograft rejection in primates. This apparently clinically applicable regimen is promising for human renal transplant trials. PMID- 15816884 TI - Activation of pro-inflammatory and anti-inflammatory cytokines in host organs during chronic allograft rejection: role of endothelin receptor signaling. AB - This study investigated whether allograft rejection is associated with local inflammatory activation in host organs and whether endothelin ET(A) receptor signaling is involved. Expression of IL-1beta, IL-1ra, IL-6, IL-10 and TNF-alpha was investigated in host liver, lung and native heart in a rat model of chronic rejection 8 weeks after heterotopic cardiac transplantation in the absence of immunosuppression. In the presence of rejection, circulating levels of cytokines increased, while tissue level activation was dependent on the organ involved. Similarly, tissue-specific regulatory patterns were observed regarding transcriptional activation. Although chronic ET(A) receptor blockade did not reduce transplant vasculopathy or tissue protein expression, treatment had pronounced effects on plasma levels and transcriptional regulation of chemokines. These data provide evidence for distinct pro-inflammatory local activation in host organs during chronic rejection and suggest a role for ET(A) receptors contributing to regulation of cytokine plasma levels and transcriptional activity. PMID- 15816885 TI - Incidence of C4d stain in protocol biopsies from renal allografts: results from a multicenter trial. AB - C4d staining of renal allografts is regarded as an in situ marker of active humoral rejection. Few data are available about the incidence of C4d deposition in protocol biopsies compared to indication biopsies. To evaluate whether center specific factors influence the incidence of C4d detection, we performed a multicenter study. From three European centers, 551 protocol and 377 indication biopsies were reclassified according to the updated Banff criteria and stained for C4d. C4d results were recorded as diffuse or focal positive and statistically correlated to clinical parameters, morphology and graft survival. In the protocol biopsies, a diffuse C4d stain was found in 2.0%, and a focal stain in 2.4%. In indication biopsies, 12.2% were diffusely and 8.5% focally C4d positive (protocol:indication p < 0.0001). The incidence of C4d deposition varied significantly between centers, attributable to variable numbers of presensitized patients with more C4d positive indication and protocol biopsies. Diffuse and focal C4d stain correlated with morphology of humoral rejection in protocol as well as in indication biopsies. Protocol biopsies show a significantly lower incidence of C4d deposition than indication biopsies. Subclinical C4d detection in protocol biopsies had no significant impact on allograft survival in our series. PMID- 15816886 TI - Obesity in living kidney donors: clinical characteristics and outcomes in the era of laparoscopic donor nephrectomy. AB - Acceptance of obese individuals as living kidney donors is controversial related to possible increased risk for surgical complications and concern that obesity may contribute to long-term renal disease. We retrospectively examined 553 consecutive hand-assisted laparoscopic living kidney donations between October 1, 1999 and April 1, 2003. We stratified donors into quartiles by baseline body mass index (BMI) assessing perioperative complications and 6-12 months post-donation metabolic and renal function. Compared to BMI <25 kg/m(2), high BMI donors (> or =35 kg/m(2)) had slightly longer operative times (mean increase 19 min), more overall perioperative complications (mostly minor wound complications), yet the same low rate of major surgical complications (conversion to open and re operation) and similar length-of-stay (2.3 vs. 2.4 days). At 6-12 months after donation (mean 11 months), renal function and microalbuminuria did not differ with BMI. These results suggest that laparoscopic donor nephrectomy is generally safe in selected obese donors and does not result in a high rate of major perioperative complications. Obese donors have higher baseline cardiovascular risk and warrant risk reduction for long-term health. While early results are encouraging, we advocate careful study of obese donors and do not support their widespread use until longer follow-up is available. PMID- 15816887 TI - Clinical utility of cytomegalovirus (CMV) serology testing in high-risk CMV D+/R- transplant recipients. AB - Late-onset cytomegalovirus (CMV) disease is a significant problem in D+/R- solid organ transplant (SOT) patients who receive antiviral prophylaxis. We assessed the clinical utility of CMV IgG and IgM serology testing for predicting late onset CMV disease. We evaluated 352 D+/R- transplant recipients who participated in a trial comparing 100 days of ganciclovir versus valganciclovir prophylaxis. CMV serology was assessed on day 28, 56, 100, and 6 and 12 months post transplant. IgG seroconversion occurred in 26.9% of patients by day 100, and in 63.4% and 75.3% by 6 and 12 months, respectively. IgM seroconversion occurred in 8.3%, 41.8% and 54.9% by day 100, month 6 and month 12, respectively. Seroconversion by day 100 (end of prophylaxis) was not predictive of subsequent CMV disease (CMV disease 13.3% if seropositive vs. 17.8% if seronegative; p = NS). However, at 6 months post-transplant, IgG serostatus was predictive of subsequent CMV disease between month 6 and 12 (CMV disease 1.3% if seropositive vs. 10.0% if seronegative; p = 0.002). In D+/R- patients, CMV serology testing is for the most part not clinically useful for predicting subsequent disease. However, seroconversion by 6 months may be useful for identifying patients at risk of late-onset CMV disease. PMID- 15816888 TI - A comparison of long-term graft survival rates between the first and second donor kidney transplanted--the effect of a longer cold ischaemic time for the second kidney. AB - Prolonged cold ischaemic time (CIT) is associated with delayed initial graft function and may also have a negative impact on long-term graft outcome. We carried out a study comparing the long-term graft survival rates between those recipients who received the first of a pair of donor kidneys versus the recipient of the second graft. Adult kidney transplant recipients who received one of a pair of donor kidneys at our institution between 1989-1995 were included. All recipients received a cyclosporin based immunosupression regimen. Graft survival rates were compared between the 2 groups at 1-, 3-, 5- and 10-year intervals. A total of 520 renal transplant grafts were included in this study. Mean donor age was 35.4 years. Groups were similar for recipient age, gender, number of HLA mismatches, transplant number for that patient and percentage PRA. CIT was the only variable that was significantly different between the two groups; mean of 19.93 h in the first group compared to 25.65 h in the second group. Graft survival rates for the first kidney were significantly better than the second kidney-graft survival at 1 year 88.5% versus 84.7%, at 3 years 81.8% versus 76.7%, at 5 years 72.2% versus 64.9% and at 10 years 55.2% versus 40% (p = 0.012). Patient survival rates were similar in both groups. In our experience, the long-term graft survival rates are significantly better for the first kidney transplanted compared to the second kidney. PMID- 15816889 TI - Significance of CT attenuation value in liver grafts following right lobe living donor liver transplantation. AB - In adult living-donor liver transplantation (LDLT), the assessment of the allograft functional reserve is important for adequate graft regeneration. From March 2002 to December 2003, 30 adult recipients underwent right lobe LDLT. Mean CT attenuation values (CT-AVs) in the graft were measured on unenhanced CT for 6 months after LDLT. The histological features of the graft parenchyma were evaluated with post-operative liver biopsy specimens. Mean CT-AVs after LDLT were decreased significantly from the pre-operative values, recovered to over 60 HU within 6 months. There was a positive linear correlation between the CT-AVs and the receptor index (LHL15) in technetium-99m-diethylenetriaminepenta-acetic acid galactosyl-human serum albumin ((99m)Tc-GSA) liver scintigraphy (r = 0.803, p = 0.005). The recipients were divided into two groups according to the CT-AV at one post-operative week (group H; > or =55HU, group L; <55HU). The low CT-AVs, under 55 HU, in group L were prolonged for 3 months compared with those in group H (p < 0.05). The 1-year cumulative survival rate was 94.7% and 45.5% in groups H and L, respectively (p = 0.014). Histological findings revealed that the parenchymal damage was severe in the grafts with low CT-AVs. The CT-AVs in the grafts may be a useful parameter for assessing the allograft functional reserve. PMID- 15816890 TI - Rapamycin successfully treats post-transplant autoimmune hepatitis. AB - Rapamycin (Rapa), one of the newer immunosuppressants has been found to control and prevent autoimmune features in animal models. This is the first report describing the successful control of post-transplant autoimmune hepatitis (AIH) with Rapa. Post-transplant AIH is diagnosed in the presence of raised transaminases, elevated immunoglobulin G, presence of autoantibodies and histologic changes consistent with AIH on liver biopsy. It may represent a recurrence of the original AIH that led to transplantation or present as a de novo AIH after liver transplant. Post-transplant AIH has conventionally been treated with Prednisolone (Pred) and Azathioprine (AZA). In this report, tailoring of immunosuppression after diagnosis of post-transplant AIH is described with special emphasis on those treated successfully with Rapa. Fifteen of 21 patients responded to treatment with an increase in dose of Pred and addition of AZA or Mycophenolate Mofetil (MMF) to calcineurin inhibitor. Five non responders and one other patient with post-transplant AIH were treated with addition of Rapa. All six responded to treatment but drug was withdrawn in one patient. Adverse events were minimal. Rapa may prove to be an important addition in the control of autoimmune liver disease. PMID- 15816891 TI - Consequences of eliminating HLA-B in deceased donor kidney allocation to increase minority transplantation. AB - HLA matching contributes to the disparity in Caucasian compared to minority kidney transplantation. HLA-B locus matching was eliminated from kidney allocation to shift a projected 166 organs from Caucasians to minorities annually. This study estimated the economic and quality-of-life impact of this policy. Cost-effectiveness analysis was performed using a Markov model. Data from the United States Renal Data System (USRDS) were used to estimate race-specific outcomes, patient and graft survival, quality-adjusted life years (QALYs) and medical costs in U.S. dollars. The greatest benefit is expected in African Americans, with expected savings of US 7.5 million dollars and 243 QALYs. Smaller cost and QALY benefits are seen in other minority groups. In Caucasians, a loss of 7.0 million dollars and a decrease of 967 QALYs are expected with the shift of organs. Overall, this policy is expected to save US 5400 dollars for each QALY that is lost. The same increase in minority transplantation would be expected from increasing Caucasian donation rates by 5.5%, or African-American donation by 29.0%, each producing large cost savings and QALY gains. Policies to increase minority transplants by increasing donation rates may prove more cost effective than the elimination of HLA-B matching from deceased donor kidney allocation. PMID- 15816892 TI - Functional status and quality of life in patients surviving 10 years after lung transplantation. AB - Although many lung allograft recipients achieve long-term survival, there is a lack of published data regarding these patients' functional status and quality of life (QoL). We evaluated all 10-year survivors at our institution and, utilizing the SF-36 questionnaire, compared their QoL to population normative and chronic illness data. Twenty-eight (29%) of 96 patients survived > or =10 years following 11 single, 6 bilateral and 11 heart-lung procedures. At the most recent evaluation, median FEV(1) in single and double lung recipients was predicted to be 54% and 74%, respectively. Five (18%) patients had BOS score 0, 13 (46%) BOS 1, 5 (18%) BOS 2 and 5 (18%) BOS 3 and median time to BOS was 7 years. Four (14%) patients required renal replacement therapy. Three patients (11%) developed symptomatic osteoporosis, 2 (7%) post-transplant lymphoma and 1 (4%) an ischaemic stroke. Scores for physical function, role-physical/emotional and general health, but not mental health and bodily pain, were significantly lower compared to normative and chronic illness data. Energy and social-function scores were significantly lower than normative data alone. Long-term survival after lung transplantation is characterized by an absence or delayed development of BOS, low iatrogenic morbidity and preserved mental, but reduced physical health status. PMID- 15816893 TI - Hospital-independent organ recovery from deceased donors: a two-year experience. AB - Early experience with deceased donor (DD) organ recovery outside of the hospital setting was found to be safe, efficient and cost effective. A 2-year experience under current practice protocols implemented to further process improvements is now reviewed. From December 1, 2001 to December 31, 2003, 123 criteria eligible DDs were transferred from local and regional hospitals to the Mid-America Transplant Services (MTS) facility for organ and tissue recovery. In this retrospective analysis, outcome comparisons were made with 79 conventional hospital-based recoveries. Compared to hospital recoveries, MTS facility recoveries were associated with significantly reduced critical care unit time (819 vs. 502 min), time to cross-clamp following brain death (966 vs. 731 min), operating room delay (54 vs. 9 min) and a trend toward reduced organ cold ischemia times which reached significance for heart and lungs when compared to regional hospital recoveries (147 vs. 221 and 192 vs. 327 min). MTS facility recovery afforded substantial cost savings over local and regional hospital recoveries (US 6,690 dollars and US 5,452 dollars per donor, respectively). The current practice of DD recovery at the MTS facility was applicable for most recoveries, improved process efficiency, and afforded substantial cost savings without donor compromise. PMID- 15816894 TI - A comparison of tacrolimus and cyclosporine in liver transplantation: effects on renal function and cardiovascular risk status. AB - A retrospective chart review of 1065 consecutive liver allograft recipients in 11 centers from January 1997 to September 1998 was performed. Patients were followed for 3 years or until graft loss. Patients received either tacrolimus (n = 594), cyclosporine (n = 450) or no calcineurin inhibitor (n = 21). Model for end-stage liver disease (MELD) scores at time of transplant were similar between the two groups. During follow-up, more patients switched from cyclosporine to tacrolimus (26.7%) than from tacrolimus to cyclosporine (12.8%; p < 0.0001). Patient and graft survival were equivalent. Corticosteroid use was more common in cyclosporine-treated patients (p < 0.00001). Patients receiving tacrolimus experienced lower serum creatinine levels at months 3 through 36 (p < 0.0001). Systolic blood pressure was lower in patients receiving tacrolimus (p < 0.001) despite a reduced requirement for anti-hypertensive agents (p < 0.0001). In addition, tacrolimus was associated with lower total cholesterol and triglyceride levels for months 3 through 24 and 3 through 12, respectively (p < 0.01), despite a reduced requirement for anti-hyperlipidemic agents. The incidence of new-onset diabetes mellitus was similar in both groups. While both calcineurin inhibitors were associated with excellent patient and graft survival, renal function, blood pressure and serum lipid levels were significantly better with tacrolimus treatment. PMID- 15816895 TI - Renal histopathological lesions after orthotopic liver transplantation (OLT). AB - Liver transplant recipients are at risk of chronic renal failure (CRF), customarily considered to be secondary to CsA/FK506 nephrotoxicity. We have examined renal biopsies from 26 liver transplant recipients with CRF. Before OLT, 5 patients had CRF, 8 were diabetic and 9 hypertensive. Renal biopsies were performed at a mean of 5 years after liver transplantation. Mean SCr was then 212 micromol/L, proteinuria was 1 g/24 h. Twelve patients were diabetic and 25 hypertensive. Histology revealed impressive renal destruction, with a mean of 45% interstitial fibrosis and 45% glomerular sclerosis. All biopsies showed severe arteriosclerosis. CRF can be attributed to four associated primary lesions: (i) specific chronic CsA/FK506 arteriolopathy; (ii) typical diabetic nephropathy; (iii) acute or chronic thrombotic microangiopathy attributed to CsA/FK506 or alpha-IFN and (iv) tubular changes related to administration of hydroxyethylstarch. At the end of the follow-up, after a mean of 6.4 years, 12 patients required dialysis, 13 had CRF and only 1 had normal renal function. Thus, CRF in OLT recipients is more complex than originally thought and should not be classified as anti-calcineurin nephrotoxicity without further investigations, including renal histology. These investigations have therapeutic potential, that is, they may lead to a more aggressive treatment of hypertension and/or diabetes. PMID- 15816897 TI - Recrudescent tobacco exposure following heart transplantation: clinical profiles and relationship with athero-thrombosis risk markers. AB - To identify tobacco recidivism among 86 heart transplant recipients who were smokers but demonstrated compliance with a smoking cessation program pre transplant, we used a questionnaire and randomly tested urine for nicotine and its by-products. In 36 patients, we also evaluated circulating levels of HS-CRP, homocysteine and MPV. Twenty-eight (32.5%) of 86 patients met our definition for tobacco exposure. In this cohort, 28 (32.5%) of 86 patients met our definition for tobacco exposure. Of these 28, 12 patients self-reported tobacco use and demonstrated biochemical verification; 14 patients demonstrated only biochemical evidence of significant tobacco exposure; 2 patients self-reported tobacco use but did not demonstrate biochemical positivity. Smoking cessation within 6 months of transplantation (r = 0.52) and time post-transplantation (r = 0.43) were independent predictors for recidivism of tobacco use, p < 0.01. No differences in HS-CRP, homocysteine and MPV levels were noted among the groups. Our investigation demonstrates a high rate of tobacco recidivism among heart transplant recipients, yet few admit to it. The adverse effects of tobacco do not appear to be directly modulated by an effect on athero-thrombotic risk markers. PMID- 15816896 TI - Kidney allograft fibrosis and atrophy early after living donor transplantation. AB - Kidney allograft failure is most often caused by chronic allograft nephropathy, a process of interstitial fibrosis (GIF) and tubular atrophy (TA). We assessed the pathology of living donor (LD) grafts compared to deceased donor (DD). Included are 321 recipients (245 LD; 76 DD) with protocol biopsies the first 2 years of transplant. In LD, GIF was present in 7%, 31%, 61% and 71% of grafts at 0, 4, 12 and 24 months. TA progressed in parallel to GIF. Compared to LD, more DD grafts had GIF at time 0 (29%, p = 0.002); thereafter the incidence of GIF was similar. In LD, GIF was associated with lower glomerular filtration rate (GFR)(1 year) (no GIF, 62 +/- 16; GIF, 49 +/- 15 mL/min/m(2) iothalamate clearance, p = 0.001) and reduced graft survival (HR = 2.2, p = 0.009). GIF in LD related to acute rejection (HR = 2.6, p = 0.01), polyoma nephropathy (OR = 4.4, p = 0.02) and lower levels of GFR 3 weeks post-transplant (HR = 0.961; p = 0.03, multivariate). However, GIF also developed in 53% of recipients lacking these covariates. Thus, GIF/TA develops in the majority of LD grafts, it is often mild but is associated with reduced function and survival. GIF frequently develops in the absence of risk factors. Lower GFR post-transplant identify patients at highest risk of GIF. PMID- 15816898 TI - To do or not to do living donor hepatectomy in Jehovah's Witnesses: single institution experience of the first 13 resections. AB - Living donor liver transplantation has come to be an acceptable alternative to deceased donor transplants. Several ethical issues related to living donation have been raised in the face of reported perioperative morbidity and mortality. We report our experience in 13 consecutive Jehovah's Witness (JW) donor hepatectomies. From June 1999 to April 2004, 13 adult JW donors underwent donor hepatectomies at the USC-University Hospital. Nine donors underwent right lobectomy with a 62% mean volume of the liver resected. Four donors underwent a left lateral segmentectomy with a mean volume of 17.8%. Cell scavenging techniques, acute normovolemic hemodilution and fractionated products were used. The mean hospital stay was 6.2 days. All donors are alive and well at a median follow-up time of 3 years and 4 months. Live liver donation can be done safely in JW population if performed within a comprehensive bloodless surgery program. PMID- 15816899 TI - Hemolytic uremic syndrome: a fatal outcome after kidney and liver transplantation performed to correct factor h gene mutation. AB - Factor H-associated hemolytic uremic syndrome (HUS) is a genetic form of thrombotic microangiopathy characterized by deficient factor H (HF-1) levels/activity and uncontrolled complement activation. The disorder mostly leads to end-stage renal disease and often recurs after kidney transplantation. We previously demonstrated that in a child with HF-1-associated HUS a simultaneous kidney and liver transplantation restored the defective HF-1 with no recurrence of the disease in the transplanted kidney. Here we describe a second childhood case of HF-1-associated HUS treated by combined kidney and liver transplant and complicated by a fatal, primary non-function of the liver graft. Graft hypoperfusion during surgery triggered ischemia/reperfusion changes and complement activation. Conceivably, as a result of defective complement regulatory potential, massive shedding of vascular heparan sulfates was documented in the transplanted liver. This might have impaired the physiological thromboresistance of vascular endothelium ending with widespread microvascular thrombosis and infarction. This case indicates that more fundamental research is needed before combined liver and kidney transplant is considered an option for children with HF-1-associated HUS. PMID- 15816900 TI - Successful outcome of progressive multifocal leukoencephalopathy in a renal transplant patient. AB - We report the case of a 47-year-old man who developed progressive multifocal leukoencephalopathy (PML) after receiving immuno-suppressive therapy for renal transplantation. The patient presented with a focal seizure and cognitive changes 5 months post-transplantation. He was found to have enhancing lesions in the parietal lobe and typical findings of PML in a brain biopsy. Immunosuppression was discontinued and the neurological symptoms gradually resolved over a period of 4 weeks. The patient is free of any neurological symptoms 36 months after the diagnosis of PML and imaging studies demonstrate resolution of the PML lesions. The patient returned to hemodialysis 3 months after immunosuppression was discontinued. We also present a review of the literature on PML in renal transplant recipients. PMID- 15816901 TI - Heart transplantation in a 68-year-old patient with senile systemic amyloidosis. AB - Senile systemic amyloidosis (SSA) results from deposition, predominantly in the heart, of amyloid fibrils derived from wild-type transthyretin (T TR) molecules. Cardiac autopsies indicate that SSA progressively increases in subjects 80 years of age and older. However, only a few cases of patients with SSA and cardiac failure have been recognized by cardiac biopsies during life. Here, we report a case of heart transplantation in a 68-year-old male patient with SSA. After cardiopulmonary resuscitation in October 1998, he underwent complete evaluation. Myocardial biopsies revealed the presence of amyloid deposition. Immunohistochemical staining of the amyloid indicated T TR. Genomic DNA analysis of the T TR exons did not result in any identification of a mutation. In 2001, heart transplantation was performed because progressive heart failure occurred. At the 1-year follow-up, no amyloid deposits were found in the donor heart. At the 2-year follow-up, the patient's physical and mental health was excellent. We conclude that heart transplantation can be an effective treatment in progressive heart failure due to SSA. PMID- 15816902 TI - Update on guidelines for prevention and management of mycobacterium tuberculosis infections after transplant. PMID- 15816903 TI - Mycophenolate sodium does not reduce the incidence of GI adverse events compared with mycophenolate mofetil. PMID- 15816905 TI - A Congressional gold medal for transplant donors and families. PMID- 15816906 TI - Preservation of renal structure and function at 2 years without calcineurin inhibitor use. PMID- 15816908 TI - Conscription of cadaveric organs: we need to start talking about it. PMID- 15816909 TI - How should we ethically select living kidney donors when they all are at risk? PMID- 15816910 TI - Abstracts of Diabetes UK Annual Professional Conference, Glasgow, United Kingdom, 20-22 April 2005. PMID- 15816911 TI - On the bottom of the deep blue sea. PMID- 15816912 TI - Quorum sensing: the power of cooperation in the world of Pseudomonas. AB - Work over the past few years has provided evidence that quorum sensing is a generic regulatory mechanism that allows bacteria to launch a unified, coordinated response in a population density-dependent manner to accomplish tasks which would be difficult, if not impossible, to achieve for a single bacterial cell. Quorum sensing systems are widespread among pseudomonads and the one of the human opportunistic pathogen Pseudomonas aeruginosa belongs to the most extensively studied cell-to-cell communication systems. In this organism, quorum sensing is highly complex and is made up of two interlinked N-acyl homoserine lactone (AHL)-dependent regulatory circuits, which are further modulated by a non AHL-related signal molecule and numerous regulators acting both at the transcriptional and post-transcriptional level. This genetic complexity may be one of the key elements responsible for the tremendous environmental versatility of P. aeruginosa. Work of the past few years showed that quorum sensing is essential for the expression of a battery of virulence factors as well as for biofilm formation in P. aeruginosa and thus represents an attractive target for the design of novel drugs for the treatment of P. aeruginosa infections. Furthermore, the cell-to-cell communication ability was also demonstrated in a number of additional pseudomonads. PMID- 15816913 TI - Crystal ball. Will we ever harness microbes to supply energy and essential elements? PMID- 15816914 TI - Crystal ball. The physiological challenge. PMID- 15816915 TI - Crystal ball. Where are all the species? PMID- 15816916 TI - Crystal ball. Between a rock and a hard place: geomicrobial electron transfer. PMID- 15816917 TI - Crystal ball. The roots of the "species" concept must be quantified. PMID- 15816918 TI - Crystal ball. The shape of microbial diversity. PMID- 15816919 TI - Crystal ball. The second coming of physics into (micro)biology. PMID- 15816920 TI - Crystal ball. In silico biology meets in situ phenomenology. PMID- 15816921 TI - Crystal ball. Getting a better picture of evolution. PMID- 15816922 TI - Crystal ball. With oceans of new data, to sink or to swim? PMID- 15816923 TI - Crystal ball. The viriosphere: the greatest biological diversity on Earth and driver of global processes. PMID- 15816924 TI - Crystal ball. Systems biology: in the broadest sense of the word. PMID- 15816925 TI - Crystal ball. The community level: physiology and interactions of prokaryotes in the wilderness. PMID- 15816926 TI - Nickel mine spoils revegetation attempts: effect of pioneer plants on two functional bacterial communities involved in the N-cycle. AB - Nickel mine spoils in New Caledonia represent an extreme environment, rich in nickel and strongly deficient in elementary elements such as carbon and nitrogen. To rehabilitate these sites, revegetation attempts are performed with endemic plant species establishing dinitrogen-fixation symbiosis (Gymnostoma webbianum and Serianthes calycina). As this biological fixation process provides the major source of available nitrogen in this extreme environment, it could be expected that nitrogen cycling would be stimulated. To study the revegetation effect on mine spoils, the effect of the two pioneer plants on the structure and activity of two functional bacterial communities involved in the N-cycle was investigated. nifH and narG genes were used as molecular markers for dinitrogen-fixers and dissimilatory nitrate reducers respectively. In order to assess the influence of the plants on both communities, nine clone libraries were constructed for each targeted gene. Libraries containing 602 and 513 nifH and narG clones, respectively, were screened by restriction fragment length polymorphism (RFLP) analysis. One hundred and forty-one and 78 representative clones from at least all RFLP families containing more than one clone were sequenced from nifH and narG clone libraries respectively. Both pioneer plants modified the diversity and activity of the two functional communities. However, distinct effects were observed depending on the plant species and the community considered. Serianthes calycina strongly selected a diazotroph phylotype and restored the potential activity of both communities. In contrast, G. webbianum selected no particular phylotype and only restored a fixing activity. PMID- 15816927 TI - Genetic diversity of marine Synechococcus and co-occurring cyanophage communities: evidence for viral control of phytoplankton. AB - Unicellular cyanobacteria of the genus Synechococcus are a major component of the picophytoplankton and make a substantial contribution to primary productivity in the oceans. Here we provide evidence that supports the hypothesis that virus infection can play an important role in determining the success of different Synechococcus genotypes and hence of seasonal succession. In a study of the oligotrophic Gulf of Aqaba, Red Sea, we show a succession of Synechococcus genotypes over an annual cycle. There were large changes in the genetic diversity of Synechococcus, as determined by restriction fragment length polymorphism analysis of a 403- bp rpoC1 gene fragment, which was reduced to one dominant genotype in July. The abundance of co-occurring cyanophage capable of infecting marine Synechococcus was determined by plaque assays and their genetic diversity was determined by denaturing gradient gel electrophoresis analysis of a 118-bp g20 gene fragment. The results indicate that both abundance and genetic diversity of cyanophage covaried with that of Synechococcus. Multivariate statistical analyses show a significant relationship between cyanophage assemblage structure and that of Synechococcus. These observations are consistent with cyanophage infection being a major controlling factor in picophytoplankton succession. PMID- 15816928 TI - Marine actinomycetes related to the "Salinospora" group from the Great Barrier Reef sponge Pseudoceratina clavata. AB - Ten strains identified as marine actinomycetes related to the "Salinospora" group previously reported only from marine sediments were isolated from the Great Barrier Reef marine sponge Pseudoceratina clavata. The relationship of the isolates to "Salinospora" was confirmed by phylogenetic analysis of 16S rRNA gene sequences. Colony morphology and pigmentation, occurrence and position of spores, and salinity requirements for growth were all consistent with this relationship. Genes homologous to beta-ketosynthase, an enzyme forming part of a polyketide synthesis complex, were retrieved from these isolates; these genes shared homology with other Type I ketosynthase genes, and phylogenetic comparison with amino acid sequences derived from database beta-ketosynthase genes was consistent with the close relationship of these isolates to the actinomycetes. Primers based on 16S rRNA gene sequences and designed for targeting amplification of members of the "Salinospora" group via polymerase chain reaction have been used to demonstrate occurrence of these actinomycetes within the sponge tissue. In vitro bioassays of extracts from the isolates for antibiotic activity demonstrated that these actinomycetes have the potential to inhibit other sponge symbionts in vivo, including both Gram-negative and Gram-positive bacteria. PMID- 15816929 TI - Diversity within cyanobacterial mat communities in variable salinity meltwater ponds of McMurdo Ice Shelf, Antarctica. AB - This study investigated the diversity of cyanobacterial mat communities of three meltwater ponds--Fresh, Orange and Salt Ponds, south of Bratina Island, McMurdo Ice Shelf, Antarctica. A combined morphological and genetic approach using clone libraries was used to investigate the influence of salinity on cyanobacterial diversity within these ecosystems without prior cultivation or isolation of cyanobacteria. We were able to identify 22 phylotypes belonging to Phormidium sp., Oscillatoria sp. and Lyngbya sp. In addition, we identified Antarctic Nostoc sp., Nodularia sp. and Anabaena sp. from the clone libraries. Fresh (17 phylotypes) and Orange (nine phylotypes) Ponds showed a similar diversity in contrast to that of the hypersaline Salt Pond (five phylotypes), where the diversity within cyanobacterial mats was reduced. Using the comparison of identified phylotypes with existing Antarctic sequence data, it was possible to gain further insight into the different levels of distribution of phylotypes identified in the investigated cyanobacterial mat communities of McMurdo Ice Shelf. PMID- 15816930 TI - Novel microbial diversity adherent to plant biomass in the herbivore gastrointestinal tract, as revealed by ribosomal intergenic spacer analysis and rrs gene sequencing. AB - It is well recognized that a dynamic biofilm develops upon plant biomass in the herbivore gastrointestinal tract, but this component of the microbiome has not previously been specifically sampled, or directly compared with the biodiversity present in the planktonic fraction of digesta. In this study, the digesta collected from four sheep fed two different diets was separated into three fractions: the planktonic phase, and the microbial populations either weakly or tightly adherent to plant biomass. The community DNA prepared from each fraction was then subjected to both ribosomal intergenic spacer analysis (RISA) and denaturing gradient gel electrophoresis (DGGE). Both types of analysis showed that dietary factors influence community structure, and that the adherent fractions produced more complex profiles. The RIS-clone libraries prepared from the planktonic and adherent populations were then subjected to restriction fragment length polymorphism (RFLP) and DNA sequence analyses, which resulted in a far greater degree of discrimination among the fractions. Although many of the sequenced clones from the adherent populations were assigned to various clusters within the low G+C Gram-positive bacteria, the clone libraries from animals consuming an all-grass diet were largely comprised of novel lineages of Clostridium, while in animals consuming the starch-containing diet, Selenomonas and Ruminococcus spp. were the dominant low G+C Gram-positive bacteria. Additionally, the libraries from hay-fed animals also contained clones most similar to asaccharolytic Clostridia, and other Gram-positive bacteria that specialize in the transformation of plant phenolic compounds and the formation of cinnamic, phenylacetic and phenylpropionic acids. These results reveal, for the first time, the phylogeny of adherent subpopulations that specialize in the transformation of plant lignins and other secondary compounds, which potentiate polysaccharide hydrolysis by other members of the biofilm. PMID- 15816931 TI - Flux and turnover of fixed carbon in soil microbial biomass of limed and unlimed plots of an upland grassland ecosystem. AB - The influence of liming on rhizosphere microbial biomass C and incorporation of root exudates was studied in the field by in situ pulse labelling of temperate grassland vegetation with (13)CO(2) for a 3-day period. In plots that had been limed (CaCO(3) amended) annually for 3 years, incorporation into shoots and roots was, respectively, greater and lower than in unlimed plots. Analysis of chloroform-labile C demonstrated lower levels of (13)C incorporation into microbial biomass in limed soils compared to unlimed soils. The turnover of the recently assimilated (13)C compounds was faster in microbial biomass from limed than that from unlimed soils, suggesting that liming increases incorporation by microbial communities of root exudates. An exponential decay model of (13)C in total microbial biomass in limed soils indicated that the half-life of the tracer within this carbon pool was 4.7 days. Results are presented and discussed in relation to the absolute values of (13)C fixed and allocated within the plant soil system. PMID- 15816932 TI - Phenotypic characterization of Rice Cluster III archaea without prior isolation by applying quantitative polymerase chain reaction to an enrichment culture. AB - A so far uncultured member of the Euryarchaeota was enriched from an anoxic riparian soil and phenotypically characterized using quantitative polymerase chain reaction (qPCR; "real-time PCR"). The microorganism is related to the Thermoplasmatales and belongs to Rice Cluster III (RC-III). Enrichment cultures utilized yeast extract (YE) by transiently accumulating acetate as major fermentation product, which was subsequently converted to methane. The abundance of RC-III archaea within the enrichment cultures was quantified by analysis of the terminal restriction fragment length polymorphism (T-RFLP) and by qPCR. We developed qPCR assays targeting the 16S rRNA genes (16S rDNA) specific for RC-III as well as for the Archaea in general. The enrichment cultures consisted of a mixed methanogenic community of Bacteria and Archaea, the latter consisting of up to 60% of members of RC-III. The other archaea belonged to Methanosarcinaceae, Methanomicrobiaceae and Methanobacteriaceae. The enriched RC-III archaea were represented by two sequences (LL25A, LL37A) that were highly similar to each other and to those detected in the soil inoculum (>98% similarity). However, the 16S rDNA copy numbers of RC-III archaea were about 1000-fold lower than those of Bacteria. Nevertheless, we were able to estimate growth parameters and physiological properties of one of the enriched RC-III archaea (LL25A) by measuring the increase of 16S rDNA copy numbers specific for this group under different growth conditions. The enriched RC-III archaeon grew optimally at temperatures between 20 and 30 degrees C and neutral pH using YE, meat extract, peptone or tryptone under anoxic conditions. Doubling time was approximately 3 days. No proliferation was detected on carbohydrates, amino acids, fatty acids, glycerol, alcohols, aromatic compounds, purine and pyrimidine bases or pyruvate. Various exogenous electron acceptors (e.g. ferric iron, S(0)) did not support growth on YE. Proliferation of the enriched RC-III archaeon was hardly affected by the antibiotics ampicillin, kanamycin and streptomycin. These findings suggest that the enriched archaeon is a mesophilic anaerobe, which can grow heterotrophically on peptides. Further enrichment on peptone and kanamycin eventually allowed the microscopic detection of coccoid cells stained by fluorescence in situ hybridization (FISH). PMID- 15816933 TI - Biomineralization of endolithic microbes in rocks from the McMurdo Dry Valleys of Antarctica: implications for microbial fossil formation and their detection. AB - In some zones of Antarctica's cold and dry desert, the extinction of cryptoendolithic microorganisms leaves behind inorganic traces of microbial life. In this paper, we examine the transition from live microorganisms, through their decay, to microbial fossils using in situ microscopy (transmission electron microscopy, scanning electron microscopy in back-scattered electron mode) and microanalytical (energy dispersive X-ray spectroscopy) techniques. Our results demonstrate that, after their death, endolithic microorganisms inhabiting Commonwealth Glacier sandstone from the Antarctica McMurdo Dry Valleys become mineralized. In some cases, epicellular deposition of minerals and/or simply filling up of empty moulds by minerals leads to the formation of cell-shaped structures that may be considered biomarkers. The continuous deposition of allochthonous clay minerals and sulfate-rich salts fills the sandstone pores. This process can give rise to microbial fossils with distinguishable cell wall structures. Often, fossilized cell interiors were of a different chemical composition to the mineralized cell walls. We propose that the microbial fossil formation observed was induced by mineral precipitation resulting from inorganic processes occurring after the death of cryptoendolithic microorganisms. Nevertheless, it must have been the organic template that provoked the diffusion of mineral elements and gave rise to their characteristic distribution pattern inside the fossilized cells. PMID- 15816934 TI - Adult non-biting midges: possible windborne carriers of Vibrio cholerae non-O1 non-O139. AB - Vibrio cholerae is a waterborne bacterium native to the aquatic environment. There are over 200 known serogroups yet only two cause cholera pandemics in humans. Direct contact of human sewage with drinking water, sea-born currents and marine transportation, represent modes of dissemination of the bacteria and thus the disease. The simultaneous cholera outbreaks that occur sometimes in distant localities within continental landmasses are puzzling. Here we present evidence that flying, non-biting midges (Diptera; Chironomidae), collected in the air, carry viable non-O1 non-O139 serogroups of V. cholerae. The association of V. cholerae with chironomid egg masses, which serve as a V. cholerae reservoir, was further confirmed. In simulated field experiments, we recorded the transfer of environmental V. cholerae by adult midges from the aquatic environment into bacteria-free water-pools. In laboratory experiments, flying adult midges that emerged from V. cholerae (O1 or O139) contaminated water transferred the green fluorescent protein (GFP)-tagged pathogenic bacteria from one laboratory flasks to another. Our findings show that aerial transfer by flying chironomids may play a role in the dissemination of V. cholerae in nature. PMID- 15816935 TI - Viruses, prokaryotes and DNA in the sediments of a deep-hypersaline anoxic basin (DHAB) of the Mediterranean Sea. AB - Viral and prokaryote abundance were investigated in a deep-hypersaline anoxic basin of the Eastern Mediterranean Sea (DHAB Atalante basin at c. 3000 m depth). This system was compared with two nearby deep-sea sites characterized by oxic conditions. Viral abundance and virus to prokaryote abundance ratio in hypersaline anoxic sediments displayed values close to those reported in oxic sites. The analysis of vertical profiles of viral abundance in the Atalante basin revealed the lack of significant changes with depth in the sediment, suggesting that benthic viruses in these anoxic and hypersaline conditions are preserved or resistant to decay. The anoxic basin displayed also very high concentrations of labile organic components (proteins and lipids) and extracellular DNA. These findings suggest that the DHAB sediments represent a reservoir for long-term preservation of benthic viruses and nucleic acids. PMID- 15816936 TI - Microbial diversity of benthic mats along a tidal desiccation gradient. AB - We investigated the influence of desiccation frequency, indicated by tidal position, on microbial community structure, diversity and richness of microbial mats. We independently characterized cyanobacterial, bacterial and archaeal communities, and their spatial variability for two distinct microbial mat systems: subtidal hypersaline mats and intertidal sand flat mats. Community fingerprints based on 16S rDNA were obtained via denaturing gradient gel electrophoresis using polymerase chain reaction primers specific for each group. Fingerprints for all three groups were consistently similar [> or =85% according to Weighted Pair Group with Arithmetic Mean (WPGMA) analysis] along a 1-km-long transect in subtidal mats. Here, pair-wise comparison analysis yielded minimal variation in diversity and richness for all groups. Fingerprints of three sites along an intertidal transect were heterogenous (> or =32% similarity according to WPGMA analysis) with clear shifts in community structure in all three microbial groups. Here, all groups exhibited statistically significant decreases in richness and diversity with tidal height (as desiccation frequency increases). Regression analysis yielded a strong correlation between diversity or richness estimates and position along the tidal gradient, for both Archaea and Bacteria, with Cyanobacteria exhibiting a weaker correlation. These results suggest that desiccation frequency can shape the structure of microbial mat communities, with Archea being least tolerant and Cyanobacteria most tolerant. PMID- 15816939 TI - Epileptic seizures and epilepsy: definitions proposed by the International League Against Epilepsy (ILAE) and the International Bureau for Epilepsy (IBE). AB - The International League Against Epilepsy (ILAE) and the International Bureau for Epilepsy (IBE) have come to consensus definitions for the terms epileptic seizure and epilepsy. An epileptic seizure is a transient occurrence of signs and/or symptoms due to abnormal excessive or synchronous neuronal activity in the brain. Epilepsy is a disorder of the brain characterized by an enduring predisposition to generate epileptic seizures and by the neurobiologic, cognitive, psychological, and social consequences of this condition. The definition of epilepsy requires the occurrence of at least one epileptic seizure. PMID- 15816940 TI - Hippocampal melatonin receptors modulate seizure threshold. AB - PURPOSE: The pineal hormone melatonin has been shown to enhance hippocampal excitability. We therefore investigated whether inactivation of hippocampal melatonin receptors affects behavioral seizures. METHODS: Intrahippocampal infusions were performed in rats to study the effect of different melatonin receptor antagonists on behavioral activity, EEG, and seizure susceptibility. Experiments were conducted at 2 times of the day that coincided with the peak and trough of the daily melatonin rhythm. RESULTS: Local infusion of the Mel(1b) receptor antagonist 4-phenyl-2-propionamidotetralin (4-P-PDOT) into the hippocampus, but not the overlying neocortex, significantly increased seizure latency and in some cases provided complete protection against seizure development. In addition, 4-P-PDOT suppressed open field activity and hippocampal EEG amplitude. The mixed Mel(1a)/Mel(1b) receptor antagonist luzindole also increased seizure latency but to a lesser degree than 4-P-PDOT. The behavioral effects of Mel(1b) receptor inhibition were comparable to those of the gamma aminobutyric acid (GABA)(A) receptor agonist muscimol and were observed during the dark phase (2400-0200 h) but not the light phase (1200-1400 h) of the daily photocycle. The anticonvulsant effect of intrahippocampal infusion of 4P-P-DOT was blocked by coadministration of the GABA(A) antagonist bicuculline. CONCLUSIONS: Our results suggest that nocturnal activation of hippocampal Mel(1b) receptors depresses GABA(A) receptor function in the hippocampus and enhances seizure susceptibility. PMID- 15816941 TI - Topiramate inhibits the initiation of plateau potentials in CA1 neurons by depressing R-type calcium channels. AB - PURPOSE: Cholinergic-dependent plateau potentials (PPs) are intrinsically generated conductances that can elicit ictal-type seizure activity. The aim of this study was to investigate the actions of topiramate (TPM) on the generation of PPs. METHODS: We used whole-cell patch-clamp recordings from CA1 pyramidal neurons in rat hippocampal slices to examine the effects of TPM on the PPs. RESULTS: In current-clamp mode, action potentials evoked PPs after cholinergic receptor stimulation. Therapeutically relevant concentrations of TPM (50 microM) depressed the PPs evoked by action potentials. Surprisingly, in voltage-clamp mode, we discovered that the cyclic nucleotide-gated (CNG) current that underlies PP generation (denoted as I(tail)) was not depressed. However, significantly longer depolarizing voltage steps were required to elicit I(tail). This suggested that the calcium entry trigger for evoking PPs was depressed by TPM and not I(tail) itself. TPM had no effect on calcium spikes in control conditions; however, TPM did reduce calcium spikes after cholinergic-receptor stimulation. We recently found that R-type calcium spikes are enhanced by cholinergic-receptor stimulation. Therefore we isolated R-type calcium spikes with a cocktail containing tetrodotoxin, omega-conotoxin MVIIC, omega-conotoxin-GVIA, omega agatoxin IVA, and nifedipine. R-type calcium spikes were significantly depressed by TPM. We also examined the effects of TPM on recombinant Ca(V)2.3 calcium channels expressed in tsA-201 cells. TPM depressed currents mediated by Ca(V)2.3 subunits by a hyperpolarizing shift in steady-state inactivation. CONCLUSIONS: We have found that TPM reduces ictal-like activity in CA1 hippocampal neurons through a novel inhibitory action of R-type calcium channels. PMID- 15816942 TI - Effects of status epilepticus early in life on susceptibility to ischemic injury in adulthood. AB - PURPOSE: Status epilepticus (SE) commonly occurs in children, whereas ischemic stroke is the most frequent neurologic insult in adults. The purpose of this study was to determine the effect of SE induced in immature (15 days old; PN15) male rats, on susceptibility to subsequent transient focal cerebral ischemia induced in adulthood. METHODS: SE was induced by flurothyl ether (FE) or kainic acid (KA). Rats that did not develop seizures after FE or KA served as controls. Five weeks later, the now-adult rats were subjected to middle cerebral artery occlusion (MCAo) for 1 or 2 h by using the intraluminal filament technique. The extent of the infarct volume was evaluated 24 h later. RESULTS: In rats submitted to 1-h-long FE-SE, the volume of infarction was significantly reduced compared with that in rats exposed to FE without SE. Longer duration of FE-SE was acutely lethal. KA-SE induced prolonged behavioral SE (156 +/- 17.5 min). In these rats, the volume of infarction was significantly larger compared with that in rats that did not show any electrographic seizures after KA administration. Comparison of FE and KA groups revealed that differences in the size of infarction were confined into cortical areas served by the MCA. Neither type of SE induced any obvious histologic changes in these neocortical regions before stroke induction. CONCLUSIONS: Early in life, SE can influence the outcome of a subsequent focal ischemic insult in adulthood. The extent of the infarct is related to the duration and cause of SE. Prolonged SE induced by KA worsens the outcome, whereas FE-SE has a neuroprotective effect. PMID- 15816943 TI - A 5-month period of epilepsy impairs spatial memory, decreases anxiety, but spares object recognition in the lithium-pilocarpine model in adult rats. AB - PURPOSE: In temporal lobe epilepsy (TLE), interictal behavioral disorders affect patients' quality of life. Therefore we studied long-term behavioral impairments in the lithium-pilocarpine (li-pilo) model of TLE. METHODS: Eleven li-pilo adult rats exhibiting spontaneous recurrent seizures (SRSs) during 5 months were compared with 11 li-saline rats. Spatial working memory was tested in a radial arm maze (RAM), anxiety in an elevated plus-maze (EPM), and nonspatial working memory in an object-recognition paradigm. Neuronal loss was assessed on thionine brain sections after behavioral testing. RESULTS: In the RAM, the time to complete each session and the number of errors per session decreased over a 5-day period in li-saline rats but remained constant and significantly higher in li pilo rats. In the EPM, the number of entries in and time spent on open arms were significantly higher in li-pilo than li-saline rats. In the object-recognition task, the two groups exhibited a comparable novelty preference for the new object. Neuronal loss reached 47-90% in hilus, CA1, amygdala, and piriform and entorhinal cortex. CONCLUSIONS: In li-pilo rats having experienced SRS for 5 months, performance in the object-recognition task is spared, which suggests that object discrimination remains relatively intact despite extensive damage. Neuronal loss in regions mediating memory and anxiety, such as hippocampus, entorhinal cortex, and amygdala, may relate to impaired spatial orientation and decreased anxiety. PMID- 15816944 TI - Electrical and chemical long-term depression do not attenuate low-Mg2+-induced epileptiform activity in the entorhinal cortex. AB - PURPOSE: Low-frequency electrical and magnetic stimulation of cortical brain regions has been shown to reduce cortical excitability and to decrease the susceptibility to seizures in humans and in vivo models of epilepsy. The induction of long-term depression (LTD) or depotentiation of a seizure-related long-term potentiation has been proposed to be part of the underlying mechanism. With the low-Mg(2+)-model of epilepsy, this study investigated the effect of electrical LTD, chemical LTD, and depotentiation on the susceptibility of the entorhinal cortex to epileptiform activity. METHODS: The experiments were performed on isolated entorhinal cortex slices obtained from adult Wistar rats and mice. With extracellular recording techniques, we studied whether LTD induced by (a) three episodes of low-frequency paired-pulse stimulation (3 x 900 paired pulses at 1 Hz), and by (b) bath-applied N-methyl-D-aspartate (NMDA, 20 microM) changes time-to-onset, duration, and frequency of seizure-like events (SLEs) induced by omitting MgSO(4) from the artificial cerebrospinal fluid. Next we investigated the consequences of depotentiation on SLEs themselves by applying low-frequency stimulation after onset of low-Mg(2+)-induced epileptiform activity. RESULTS: LTD, induced either by low-frequency stimulation or by bath applied NMDA, had no effect on time-to-onset, duration, and frequency of SLEs compared with unconditioned slices. Low-frequency stimulation after onset of SLEs did not suppress but induced SLEs that lasted for the time of stimulation and were associated with a simultaneous increase of the extracellular K(+) concentration. CONCLUSIONS: Our study demonstrates that neither conditioning LTD nor brief low-frequency stimulation decreases the susceptibility of the entorhinal cortex to low-Mg(2+)-induced epileptiform activity. The present study does not support the hypothesis that low-frequency brain stimulation exerts its anticonvulsant effect via the induction of LTD or depotentiation. PMID- 15816945 TI - Seizure frequency and cortisol and dehydroepiandrosterone sulfate (DHEAS) levels in women with epilepsy receiving antiepileptic drug treatment. AB - PURPOSE: Hormonal changes occur in epilepsy because of seizures themselves and of antiepileptic drug (AED) effects on steroid production, binding, and metabolism. Conversely, steroids may influence neuron activity and excitability by acting as neuroactive steroids. This cross-sectional observational study aimed to evaluating cortisol and dehydroepiandrosterone sulfate (DHEAS) levels in female epilepsy patients with different disease severity, as assessed by a seizure frequency score (SFS). METHODS: Morning serum levels of cortisol and DHEAS were assayed in 113 consecutive women, aged 16 to 47 years, with varied epilepsy syndromes, receiving mono- or polytherapy with enzyme-inducing and/or noninducing antiepileptic drugs (AEDs). Hormonal data were correlated with clinical parameters (age, body mass index, epilepsy syndrome, disease onset and duration, SFS, AED therapy, and AED serum levels) and compared with those of 30 age-matched healthy women. RESULTS: In epilepsy patients, cortisol levels and cortisol-to DHEAS ratios (C/Dr) were significantly higher, whereas DHEAS levels were significantly lower than those in controls. Patients with more frequent seizures showed higher cortisol and C/Dr values and lower DHEAS levels than did those with rarer or absent seizures during the previous 6 months. SFS mainly explained the increase of cortisol levels and C/Dr in patients with more active disease. Changes in DHEAS levels correlated with SFS and epilepsy syndrome, as well as with AED treatments and ages. CONCLUSIONS: Women with more frequent seizures had alterations of their adrenal steroids characterized by an increase of cortisol and a decrease of DHEAS levels. Such hormonal changes might be relevant in seizure control and in patient health. PMID- 15816946 TI - A clinical and EEG study on idiopathic partial epilepsies with evolution into ESES spectrum disorders. AB - PURPOSE: Questioning the presence of any possible prognostic predictors, this study includes a long-term follow-up of clinical and EEG characteristics of 16 patients with idiopathic partial epilepsy (IPE) who subsequently developed epilepsy with electrical status epilepticus during slow sleep (ESES) spectrum disorders. METHODS: Epilepsy, cognitive and behavioral parameters, and waking and non-rapid eye movement (NREM) EEG data were evaluated and scored for initial stage (i.e., IPE stage), preESES, ESES, and ESES remission periods, individually, on a chronologic basis. Data from 25 healthy subjects who had had IPE at the appropriate ages served for comparison with the patients' data during the IPE stage. RESULTS: Results revealed a higher incidence in seizure frequency and variability in the ESES group and a resistance to a single antiepileptic drug (AED), as compared with controls, during the IPE stage. Mean age at onset of epilepsy was younger in the ESES group versus controls (5.5 and 7.3 years, respectively). At least one of the premonitory clinical features for development of ESES [an increase in the seizure frequency and/or addition of new types of seizures (93%), appearance of cognitive and/or behavioural changes (81.2%), or a progression in EEG abnormalities (66%)] was present in all patients. Epilepsy remitted in patients within the ESES spectrum at a similar age as in controls in 81.2%, as ESES findings in the EEG disappeared by age 13 years in 94%. Seizure prognosis proved to be the most favorable among the questioned parameters. CONCLUSIONS: An increase in seizure frequency or development of new seizure types, a deviance in behavior or decrease in cognitive performance, or a spreading tendency of the previously focal abnormalities in control EEGs may be premonitory features of a developing ESES and necessitate close follow-ups with sleep EEGs in children with IPEs. PMID- 15816947 TI - Somatosensory processing is impaired in temporal lobe epilepsy. AB - PURPOSE: Growing evidence suggests that temporal lobe epilepsy (TLE) is a network disease. In this view, the seizure focus may produce measurable deficits in specific cortical functions. METHODS: A tactile grating orientation (GrOr) discrimination task associated with parietal lobe function was administered at the index fingertip to 15 subjects with medically intractable TLE and to 19 neurologically normal controls. TLE subjects were tested bilaterally at baseline while taking their usual antiepileptic drugs (AEDs), and off AEDs during inpatient video-EEG monitoring (n = 9). Three subjects also were tested after temporal lobectomy. t Tests were used to compare baseline performance between TLE subjects and controls, and between hands ipsilateral and contralateral to side of seizure onset, with Bonferroni correction for multiple comparisons. TLE subjects' baseline thresholds were compared with those obtained off AEDs by using a repeated measures analysis of variance. RESULTS: TLE subjects were severely impaired bilaterally on the GrOr task, with mean discrimination thresholds nearly twice those of controls (p 90% seizure reduction, and 13 (34%) were improved or unchanged. Children seizure free at two-year follow-up were likely to remain seizure free. Ten-year seizure freedom was 72% in children with developmental tumors and 32% in the cortical dysplasia group. Complete resection was statistically significant for favorable outcome, and no patient with an incomplete resection was seizure free. PMID- 15816951 TI - Cognitive function in preschool children after epilepsy surgery: rationale for early intervention. AB - PURPOSE: The detrimental effect of frequent early seizures on the cognitive potential of children is a significant clinical issue. Epilepsy surgery in childhood offers a good prognosis for seizure control and improved developmental outcome. We studied the postoperative outcome and the developmental velocity after surgery and analyzed risk factors for developmental delay in 50 consecutive preschool children treated surgically for severe epilepsy at ages 3 to 7 years. METHODS: Pre- and postoperative developmental quotients (DQs) were analyzed with analysis of variance; stepwise linear regressions were performed on preoperative DQs and on a difference score between post- and preoperative DQs to determine risk factors for preoperative development and factors influencing postoperative development. RESULTS: Of the 50 patients, 70% were retarded, with IQ < 70; 16% were of average intelligence, with IQ ranging from 85 to 115. Age at seizure onset and extent of lesion were predictive variables for preoperative cognitive development. Six to 12 months after surgery (early postoperative phase), 66% were seizure free (Engel outcome class I), 26% had substantial to worthwhile seizure reduction (classes II and III), and 8% were unchanged (class IV). Forty-one (82%) children showed stable velocity of development; three children showed gains of >/=15 IQ points; three had developmental decline (loss of >/=10 IQ points), which was transient in two children; and three children moved from not assessable to assessable. At last follow-up (6 months to 10 years after surgery), 11 children showed IQ/DQ gains of >/=15 IQ points. Gains in IQ were observed only in seizure free children and were stable over time. Shorter duration of epilepsy was significantly associated with a postoperative increase in DQ. CONCLUSIONS: (a) Substantial global mental delay is common in young children treated for epilepsy with surgery; (b) In most patients, postoperative development proceeded at a stable velocity; (c) Catch-up development may occur but only in seizure-free patients; (d) Substantial cognitive losses were noted in only one child. and (e) Early seizure control stabilized developmental velocity in this patient cohort. PMID- 15816952 TI - Migrating partial seizures in infancy: expanding the phenotype of a rare seizure syndrome. AB - PURPOSE: The constellation of early-onset, unprovoked, alternating electroclinical seizures and neurodevelopmental devastation was first described by Coppola et al. We report six new patients and the prospect of a more optimistic developmental outcome. METHODS: Retrospective chart reviews were performed on six infants evaluated at the Children's Hospital of Philadelphia (five patients) and at Hershey Medical Center (one patient) who had electroclinically alternating seizures before age 6 months of age. Electroclinical characteristics and long-term follow-up were recorded. RESULTS: All had unprovoked, early-onset (range, 1 day to 3 months; mean, 25 days) intractable electroclinical seizures that alternated between the two hemispheres. Each patient underwent comprehensive brain imaging and neurometabolic workups, which were unrevealing. In all patients, subsequently intractable partial seizures developed and often a progressive decline of head circumference percentile occurred with age. Three demonstrated severe developmental delay and hypotonia. All survived, and 7-year follow-up on one patient was quite favorable. CONCLUSIONS: Our patients satisfied the seven major diagnostic criteria first described by Coppola et al. The prognosis of this rare neonatal-onset epilepsy syndrome from the original description and subsequent case reports was very poor, with 28% mortality, and the majority of survivors were profoundly retarded and nonambulatory. Our patient data validate the diagnostic criteria of this syndrome and further quantify a previously described observation of progressive decline of head circumference percentiles with age. Our data also suggest that the prognosis of this syndrome, although poor, is not as uniformly grim as the cases reported previously in the literature. PMID- 15816953 TI - Correlation of severity of FDG-PET hypometabolism and interictal regional delta slowing in temporal lobe epilepsy. AB - PURPOSE: We investigated the association of severity of hypometabolism detected by positron emission tomography (PET) with [(18)F]fluorodeoxyglucose (FDG) and persistence of interictal EEG focal slowing in patients with refractory temporal lobe epilepsy. METHODS: Eighty temporal lobes of 40 consecutive patients with intractable temporal lobe epilepsy (mean age, 43.5 years) were studied. All patients underwent video-EEG monitoring, magnetic resonance imaging (MRI), and FDG-PET. Patients with either normal MRI or with unilateral mesial temporal sclerosis, but no other structural abnormality, were included. Interictal EEG delta slowing was graded as none, infrequent (one episode or less/hour), intermediate (more than one episode/hour), or continuous. PET hypometabolism was graded as none, mild, moderate, or severe. RESULTS: The severity of temporal lobe hypometabolism with PET was significantly correlated with the amount of delta activity in the interictal EEG, independent of MRI findings (Spearman r = 0.46; p < 0.0005). CONCLUSIONS: This observation suggests related underlying pathophysiologic mechanisms for metabolic and electrical dysfunction in temporal lobe epilepsy. PMID- 15816954 TI - A child with epilepsy in whom multifocal VEPs facilitated the objective measurement of the visual field. AB - PURPOSE: We investigated whether visual field defects can be objectively evaluated by using multifocal visual evoked potential (multifocal VEP) in children with epilepsy in whom it is difficult to measure the visual field. METHODS: First, to determine normal waves in multifocal VEP, recording was performed by using a VERIS Junior Science (Mayo, Aichi, Japan) in 21 healthy children (21 eyes) consisting of 12 boys and nine girls aged 6 to 15 years (mean age, 11.4 years). Responses from eight sites in each subject were divided into four quadrants (superior and inferior temporal quadrants and superior and inferior nasal quadrants). In each quadrant, two response waves were grouped and averaged; peak latency and amplitude were used for assessment. In a child with epilepsy in whom kinetic or static perimetry was impossible, multifocal VEPs were recorded and compared with the peak latency and amplitude in the healthy subjects. RESULTS: In the 21 healthy children, no significant differences were observed in the peak latency or amplitude among four quadrants by one-way analysis of variance. Multifocal VEPs in the child with epilepsy showed abnormal waves in the right hemivisual field, which corresponded to the lesion observed with magnetic resonance imaging. CONCLUSIONS: The objective evaluation of visual field defects by using multifocal VEP may be useful in children with epilepsy in whom kinetic/static perimetry as a subjective examination is difficult. PMID- 15816955 TI - Can you predict an immediate, complete, and sustained response to the ketogenic diet? AB - PURPOSE: Although the ketogenic diet has been in use for >80 years, little agreement exists as to which patients are most likely to have dramatic, sudden, and complete seizure control. METHODS: A case-control study was performed of children with intractable epilepsy started on the ketogenic diet at our institution since June 2001. Patients with a dramatic response were defined as those becoming seizure free within 2 weeks of diet onset. These children were compared with all other patients treated with the diet over the same time period in terms of patient demographics, epilepsy characteristics, and diet parameters. RESULTS: Eighteen early, dramatic responders over a 3-year period were identified and compared with 89 patients who were not similarly improved. The absence of complex partial seizures as the predominant seizure type (0 vs. 23%; p = 0.02) correlated with this dramatic success. The presence of infantile spasms (39% vs. 20%; p = 0.09) approached significance, but all other variables did not. CONCLUSIONS: An early, dramatic response to the ketogenic diet is more likely in patients with predominant seizure types other than complex partial. It may also be more likely to occur in children who have infantile spasms. In all other patient demographics and diet parameters, an equal likelihood of similar success was found. PMID- 15816956 TI - Epilepsy and neurocysticercosis in Atahualpa: a door-to-door survey in rural coastal Ecuador. AB - PURPOSE: To determine the prevalence of epilepsy and the role of neurocysticercosis in the occurrence of epilepsy in Atahualpa. METHODS: We used a door-to-door survey to detect subjects with epileptic seizures, to collect a blood sample for determination of anticysticercal antibodies, and to evaluate social characteristics of the population, including household pig ownership. Neurologists examined suspected cases and a sample of negative individuals. Then patients with epilepsy, as well as age- and sex-matched controls, underwent a head computed tomography (CT) and a scalp EEG. RESULTS: The questionnaire was answered by 2,415 of 2,548 residents of Atahualpa, and cysticercosis serology was performed in 1,687 consenting individuals. Cysticercosis seroprevalence was 145 (8.6%) per 1,686). Neurologic examination confirmed 24 patients with epilepsy (crude prevalence, 9.9 per 1,000 population, and 10.8 per 1,000 when adjusted to the United States population). After adjustment by age, sex, and pig raising, positive serology was strongly associated with epilepsy (odds ratio (OR), 4.16; 95% confidence interval (CI), 1.6-11.2). CT findings compatible with neurocysticercosis were found in five patients with epilepsy and also were more frequent than in controls, although this did not reach statistical significance (five of 19 vs. one of 19; p = 0.125, McNemar's test). Besides these five cases, three other patients with epilepsy had positive serology (one with a normal CT and two who did not have a CT). CONCLUSIONS: Neurocysticercosis is associated with one-third of cases of epilepsy in Atahualpa and may be a major contributory factor for the excess fraction of epilepsy seen in this population. PMID- 15816957 TI - Hypersomnia in an epilepsy patient treated with levetiracetam. AB - We report a patient with focal epilepsy in whom increased sleep needs (hypersomnia) developed in the absence of subjective excessive daytime sleepiness (EDS) during an add-on treatment with levetiracetam (LEV). PMID- 15816958 TI - Simple partial seizures with hemisensory phenomena and dysgeusia: an insular pattern. AB - Insular seizures are rarely described, in part owing to the complex anatomy of this brain region. We present a patient with simple partial seizures, recorded intracranially, originating in the right insula and characterized by dysgeusia and contralateral somatosensory phenomena. This rare clinical pattern seems to be characteristic of the insula and may be undetectable with surface EEG. PMID- 15816959 TI - Emergency transcallosal resection of hypothalamic hamartoma for "status gelasticus". AB - Hypothalamic hamartomas are rare developmental malformations of the inferior hypothalamus. Typically they result in symptomatic, refractory mixed seizure disorder with trademark gelastic or laughing seizures. We present a 30-month-old boy with a hypothalamic hamartoma and a nearly 2-month history of near-continuous gelastic seizures, which we have labeled "status gelasticus." The seizures were refractory to multiple antiepileptic drugs (AEDs), and emergency transcallosal resection of the hypothalamic hamartoma was performed with resultant near immediate cessation of the seizures. At 12-month follow-up, the patient was averaging one brief gelastic seizure every 2 weeks, with a longest seizure-free period of 2 months. PMID- 15816960 TI - Leptomeningeal enhancement and enlarged choroid plexus simulating the appearance of Sturge-Weber disease in a child with tuberous sclerosis. PMID- 15816961 TI - Smoking cessation a byproduct of EEG telemetry monitoring. AB - Smoking is a common problem in epilepsy patients. The inpatient video-EEG monitoring (VEEG) unit provides a unique and conducive environment for epilepsy patients to participate actively in a smoking-cessation program. The restrictions and confinement to the telemetry bed impose a forced abstinence from smoking. It has been suggested that patients who are hospitalized may be more receptive to smoking-cessation advice. We report two patients who were successfully able to quit smoking after admission for VEEG. PMID- 15816962 TI - Valproate-induced proximal renal tubular dysfunction: clinically relevant in the severely disabled epileptic population. PMID- 15816963 TI - Variables in childhood epilepsy and scholastic underachievement. PMID- 15816965 TI - ATL in refractory epilepsy with normal MRI-volumetric criteria? PMID- 15816967 TI - Epilepsy in malaria. PMID- 15816971 TI - Is cognitive reserve applicable to epilepsy? The effect of educational level on the cognitive decline after onset of epilepsy. AB - PURPOSE: The aim of this study was to know the effect of education level (EL) on the cognitive change after onset of epilepsy. The rationale is that people with high EL may have more cognitive reserve and show resistance to damage brought about by epilepsy. METHODS: Patients were from an outpatient epilepsy clinic of a national university hospital. All received initial and repeated neuropsychological tests and the Cognitive Ability Screening Instrument (CASI), at an interval of 12 months. CASI consists of nine items, with a total score of 100. We compared the differences between the two tests, in terms of z-scores. The EL was divided into high and low with a cut-off at 12 years, and age at entry was divided with a cut-off at 38 years. Epilepsy severity was divided into intractable and nonintractable. RESULTS: A total of 64 patients completed the study, including 31 with high EL and 33 with low EL, with no differences in gender and epilepsy severity between groups. In general, patients had cognitive impairment in multiple domains. At the baseline, the effect of EL was found for all items except remote memory and orientation, whereas the effect of age was observed for remote and recent memory. At the 12-month follow-up, subjects with high EL showed deterioration in mental manipulation, whereas subjects with low EL showed improvement in verbal fluency but deterioration in attention. CONCLUSIONS: Patients with high EL have better cognitive functioning than those with low EL after epilepsy has developed, which supports the cognitive reserve theory. However, 1-year follow-up might be too short to see significant changes between groups. PMID- 15816970 TI - Callosal bisection and transcallosal secondary antiepileptogenesis. AB - More than 28,000 neuroscientists and 3,000 epileptologists gathered at their respective 2001 meetings of the Society for Neuroscience and the American Epilepsy Society. Yet only six articles, one directly and five indirectly, discussed the corpus callosum (CC). Is not this in itself a remarkable finding? Are there no mysteries left? The reality is that considerable uncertainties exist regarding the rationale for callosal bisection (CCB) that causes contrasting effects (i.e., amelioration of generalized seizure, at times leading to freedom from seizure, and intensification of postoperatively fragmented seizure, at times leading to status epilepticus). Similarly, the clinical relevance of EEG mirror focus formation, an experimentally well-established transcallosal consequence of partial cortical epileptogenesis, continues to be debated. This presentation revisits these unresolved issues (a) to gain insight into the dynamic role played by the CC in medically refractory epilepsy, and (b) to promote the development of antiepileptogenic tools that are currently unavailable. PMID- 15816972 TI - From precipitation to inhibition of seizures: rationale of a therapeutic paradigm. AB - Epileptic seizures can be triggered by both nonspecific facilitating factors such as sleep withdrawal, fever, or excessive alcohol intake, and specific reflex epileptic mechanisms. These consist of sensory or cognitive inputs activating circumscribed cortical areas or functional anatomic systems that, due to some functional instability, respond with an epileptic discharge. Interruption of seizure activity at the stage of the aura (i.e., locally restricted discharge) also can be achieved by nonspecific (e.g., relaxation or concentration techniques or vagal nerve stimulation) or by specific focus-targeted sensory or cognitive inputs. The latter, again, activate circumscribed cortical areas. Intriguingly, in some patients, the same stimulus can either precipitate or abort a seizure. The response depends on the state of cortical activation: seizure precipitation occurs in the resting condition, and seizure interruption occurs when the epileptic discharge has begun close to the activated area. These relations can be understood on the background of experimental data showing that an intermediate state of neuronal activation is a precondition for the generation of paroxysmal depolarization shifts, whereas a hyperpolarized neuron will remain subthreshold, and a depolarized neuron that already produces action potentials is not recruitable for other activity. Sensory input meeting an intermediately activated pool of potentially epileptic neurons is adequate to produce a seizure. In another condition, the same stimulus can depolarize a neuron pool in the same area sufficiently to block the further propagation of nearby epileptic activity. Understanding these interactions facilitates the development of successful nonpharmaceutical therapeutic interventions for epilepsy. PMID- 15816973 TI - The role of cognitive-motor function in precipitation and inhibition of epileptic seizures. AB - PURPOSE: To examine the effects of cognitive-motor function on EEG discharges and the neuropsychological mechanisms of seizure induction in patients sensitive to cognitive-motor tasks. METHODS: Four hundred eighty patients with epilepsies were subjected to cognitive tasking, termed "neuropsychological EEG activation (NPA)." It consisted of reading, speaking, writing, written calculation, mental calculation, and spatial construction. Furthermore, patients showing a provocative NPA effect were subjected to a detailed NPA protocol to identify the possible precipitating factors, which consisted of simple hand movements, action programming requiring hand movement, and thinking activity not requiring hand movement. RESULTS: NPA had an inhibitory effect on EEG discharges in 133 (63.9%) of 208 patients with discharges in the awake EEG. Conversely, NAP had a provocative effect in 38 (7.9%) of 480 patients. In 32 of the 38 patients, the precipitating factor was action programming. Among them, five showed a precipitating factor restricted to linguistic activity, and the remaining 27 were affected by various action-programming factors including both linguistic and praxic activities. In four of 38 patients, the precipitating factor was thinking, predominantly linguistic tasks in one patient and spatial tasks in three patients. No patient had a precipitating factor identified as motor activity. CONCLUSIONS: These results suggest that cognitive-motor function has an inhibitory effect on EEG discharges in the majority of epilepsy patients and a provocative effect in some patients, and that seizures of the patients showing a provocative NPA effect are precipitated by action programming or thinking activity. PMID- 15816975 TI - Surgical outcome of epilepsy caused by cortical dysplasia. AB - PURPOSE: Cortical dysplasia (CD) is the second most common pathologic entity in surgically treated epilepsy. To delineate its surgical outcome and prognostic factors, we performed a retrospective analysis of patients operated on at a single institute. METHODS: Between September 1994 and December 2000, 128 cases with CD were operated on at our institute. The male/female ratio was 85:43, and the mean age was 27 years (SD, 7.6 years). The mean postoperative follow-up period was 26.9 months (SD, 12.0 months). Seizure outcome and severity of CD were classified according to Engel's and Mischel's classifications, respectively. Severity of CD was mild in 69, moderate in 21, and severe in 38. Lobar distribution was temporal in 54, frontal in 48, and others in 26. Cortical resection, lobectomy, or hemispherectomy was done in all patients. Univariate analysis was done followed by multivariate analysis by using computerized statistical software. RESULTS: Postoperative seizure outcome was Engel class I in 58 (45.3%), II in 17 (13.3%), III in 16 (12.5%), and IV in 37 (28.9%). Transient postoperative complications occurred in 14 (10.9%) patients without mortality or permanent neurologic impairment. Multivariate analysis revealed severity of CD and frontal lobe resection as independent prognostic factors for worse outcome (p = 0.001 and 0.003, respectively). CONCLUSIONS: Epilepsy surgery for CD achieved 45.3% seizure-free rate with 10.9% transient postoperative complications in our institute. Worse postoperative seizure outcome was observed in cases with mild CD and frontal lobe resection. PMID- 15816974 TI - Study of antiepileptic effect of extracts from Acorus tatarinowii Schott. AB - PURPOSE: To study the antiepileptic properties of extracts from rhizomes of Acorus tatarinowii Schott (ATS). METHODS: The decoction and volatile oil were extracted from rhizomes of ATS by traditional decocting and supercritical CO(2) fluid extraction (SFE-CO(2)) methods. Maximal electroshock (MES), pentylenetetrazol (PTZ) maximal seizure, and prolonged PTZ kindling models were used to test their anticonvulsive properties. The gamma-aminobutyric acid (GABA) immunohistochemical reaction (IR) was used to study GABAergic neuron changes in the PTZ kindling model and the effects of treatment. RESULTS: Both decoction (dose; 10-20 g/kg) and volatile oil (1.25 g/kg) of ATS decreased the convulsive rate significantly in the MES model. Decoction of ATS was shown to be effective in the PTZ model with both decreased convulsive and mortality rates. The volatile oil of ATS failed to prevent seizures in the dose range tested, although prolonged seizure latency and decreased mortality were found at a dose of 1.25 g/kg. In the PTZ kindling model, GABA-IR neurons decreased obviously compared with the normal group. In the groups treated with the decoction and volatile oil, the seizure intensity decreased significantly after treatment. Increased GABA-IR neurons also were found when compared with PTZ kindling controls. Morphologic observation also showed that GABA-IR neuron damage was less severe in the drug treated groups. CONCLUSIONS: Both decoction and volatile oil extracted from the rhizome of Acorus tatarinowii Schott have anticonvulsive effects. The volatile oil is shown to be less effective for PTZ-induced convulsions. Both extracts can prevent convulsions as well as convulsion-related GABAergic neuron damage in the brain in the prolonged PTZ kindling model. PMID- 15816976 TI - Our experience with pediatric epilepsy surgery focusing on corpus callosotomy and hemispherotomy. AB - PURPOSE: In pediatric patients with intractable epilepsy, surgical intervention should be considered in terms of seizure control and prevention of neuronal deterioration. We present our experience with pediatric epilepsy surgery, focusing on corpus callosotomy and hemispherotomy. METHODS: A total of 257 pediatric patients underwent epilepsy surgeries during the last decade in our clinic. The two most common surgical procedures were corpus callosotomy and hemispherotomy. The surgical outcomes of the patients undergoing these operations were analyzed with regard to seizure outcomes and surgical complications. RESULTS: Corpus callosotomy proved to be extremely effective in controlling drop attacks, especially when the callosum was totally sectioned. Postoperative improvement of psychomotor function also was noticeable in >70% of the cases. In patients who underwent hemispherotomy, the etiology of unilateral hemispheric lesions was closely related to surgical results, with cases of hemimegalencephaly having the worst seizure outcome and most frequent surgical complications. Incomplete section of the callosum, one of the most important causes of residual seizures, was found in three cases. CONCLUSIONS: In pediatric epilepsy surgery, corpus callosotomy is very effective in abolishing drop attacks and improving postoperative psychomotor function. For hemispherotomy, cases of hemimegalencephaly had the worst seizure control and surgical complications. Incomplete section of the corpus callosum should be carefully evaluated as a cause of surgical failure. PMID- 15816978 TI - Genetics of idiopathic epilepsies. AB - PURPOSE: To search for clues to molecular genetics of common idiopathic epilepsy syndromes. Genetic defects have been identified recently in certain inherited epilepsy syndromes in which the phenotypes are similar to those of common idiopathic epilepsies. METHODS: Mutations identified as the causes of inherited idiopathic epilepsies were reviewed. RESULTS: Mutations of the genes encoding two subunits of the neuronal nicotinic acetylcholine receptor were found in autosomal dominant nocturnal frontal lobe epilepsy. Mutations of two K(+)-channel genes were identified in benign familial neonatal convulsions. Mutations of the genes encoding several subunits of the voltage-gated Na(+)-channel and gamma aminobutyric acid (GABA)(A) receptor also were identified as the underlying causes of various epilepsy syndromes, such as autosomal dominant epilepsy with febrile seizures plus, benign familial neonatal infantile seizures, and autosomal dominant juvenile myoclonic epilepsy. Mutations within the same gene may result in different epilepsy phenotypes. Thus, the Na(+) channel, GABA(A) receptor, and their auxiliaries may be involved in the pathogenesis of various types of epilepsy. Some forms of juvenile myoclonic epilepsy, idiopathic generalized epilepsy, and absence epilepsy may result from mutations of Ca(2+) channels. Mutations of the Cl(-) channel have been recently found to be associated with a certain type of epilepsy. The recent discovery that mutations of LGI1, a gene encoding a nonchannel molecule, are associated with autosomal partial epilepsy with auditory features may provide a new insight into our understanding of the genetics of idiopathic epilepsy. CONCLUSIONS: These findings suggest the involvement of brain channelopathies in the pathogenesis of certain types of idiopathic epilepsy. PMID- 15816977 TI - Genetic malformations of the cerebral cortex and epilepsy. AB - We reviewed the epileptogenic cortical malformations for which a causative gene has been cloned or a linkage obtained. X-linked bilateral periventricular nodular heterotopia (BPNH) consists of typical BPNH with epilepsy in female patients and prenatal lethality in most males. About 90% of patients have focal epilepsy. Filamin A mutations have been reported in all families and in approximately 20% of sporadic patients. A rare recessive form of BPNH also has been reported. Most cases of lissencephaly-pachygyria are caused by mutations of LIS1 and XLIS genes. LIS1 mutations cause a more severe malformation posteriorly. Most children have isolated lissencephaly, with severe developmental delay and infantile spasms, but milder phenotypes have been recorded. XLIS usually causes anteriorly predominant lissencephaly in male patients and subcortical band heterotopia (SBH) in female patients. Thickness of the band and severity of pachygyria correlate with the likelihood of developing Lennox-Gastaut syndrome. Mutations of the coding region of XLIS are found in all reported pedigrees and in 50% of sporadic female patients with SBH. Autosomal recessive lissencephaly with cerebellar hypoplasia; accompanied by severe delay, hypotonia, and seizures, has been associated with mutations of the RELN gene. Schizencephaly has a wide anatomoclinical spectrum, including focal epilepsy in most patients. Familial occurrence is rare. Initial reports of heterozygous mutations in the EMX2 gene need confirmation. Among several syndromes featuring polymicrogyria, bilateral perisylvian polymicrogyria shows genetic heterogeneity, including linkage to Xq28 in some pedigrees, autosomal recessive inheritance in others, and association with 22q11.2 deletion in some patients. About 65% of patients have severe epilepsy, often Lennox Gastaut syndrome. Recessive bilateral frontal polymicrogyria has been linked to chromosome 16q12.2-21. PMID- 15816979 TI - Development of an epilepsy comprehensive care center: a Japanese model. AB - PURPOSE: To review the requirements of a comprehensive care center for people with epilepsy. METHODS: Twenty-seven years have passed since the foundation of the Japanese Epilepsy Center in Shizuoka. The development of this center is presented as a model of an epilepsy comprehensive care center. RESULTS: Between 1926 and 1947, the Shizuoka Higashi Hospital (the former name of SMIND) served as a tuberculosis hospital. In 1975, a proposal for a special center for the care of people with epilepsy was submitted to the Japanese government. An epilepsy center (the Center) was soon built, and the tuberculosis sanatorium ended its 50-year history. The facilities of the Center include an outpatient clinic, four inpatient wards with 200 beds, a day-care center for medical rehabilitation, and classrooms for elementary and junior high school children. The Center has modern diagnostic tools such as electroencephalography (EEG), closed-circuit TV-EEG (CCTVEEG), computed tomography (CT) scan, magnetic resonance imaging (MRI), single-photon emission CT (SPECT), and magnetoencephalography (MEG). Neurosurgery for intractable seizures has been conducted at the Center since 1983. Approximately 25,000 patients with epilepsy from all over Japan have been registered since 1975. Annually, approximately 5,000 outpatients and 600 inpatients attend the Center. As of March 2002, 500 patients had received resective surgery for epilepsy. Other activities in the Center include research and specialized training of professionals, including foreign nationals, in the treatment of epilepsy. CONCLUSIONS: The experience in Shizuoka suggests that management of epilepsy should be oriented toward psychological well-being, social rehabilitation, and seizure control. PMID- 15816980 TI - Knowledge, attitudes, and perceptions of epilepsy in Asia: toward a uniform study protocol. AB - PURPOSE: Knowledge, attitudes, and perceptions of epilepsy (KAPE) among the public play a major role in determining the extent to which people with epilepsy can be integrated into their society. KAPE SURVEYS IN ASIA: Surveys conducted among Chinese in China, Taiwan, Malaysia, Singapore, and Hong Kong demonstrated similarities as well as differences in certain aspects of KAPE. These suggest that KAPE might not be uniform among other racial groups in the same or different regions of Asia. For KAPE to have usefulness to a country or community, it has to be location specific. TOWARD A UNIFORM STUDY PROTOCOL: There are several advantages of using a uniform study protocol. Collection of KAPE data could be faster for any country or community that chooses to use any existing study protocol. Intraregional, interregional, intercontinental, interethnic, intercommunity, and longitudinal comparison within any subgroup could be carried out. Sharing of common information would enhance the regional effort in improving the quality of life of people affected by epilepsy. However, survey methods need not be uniform. Regional adaptation and flexibility of a unified protocol should be encouraged. After the study, concerted local and regional efforts in epilepsy education must be carried out systematically for the general public as well as for strategic groups. CONCLUSIONS: Determination of the KAPE in different parts of Asia would help to define the magnitude of and be the first step toward alleviating social discrimination against people with epilepsy in Asia. PMID- 15816981 TI - Quality-controlled education in epileptology: experiences from Europe and possible developments for other regions. AB - As a consequence of the political system shift in Eastern Europe in the early 1990s, the new International League Against Epilepsy (ILAE) Commission on European Affairs (CEA) had to face the task of ensuring an equally good quality of epilepsy care across the whole region. A high standard of epileptologic education being the most important precondition for that, the decision was taken to make quality-controlled education a first priority. To reach this aim, an "European Epilepsy Academy" (Eurepa) was founded in 1996. Its activities comprise organization of its own educational courses, certification of other educational courses, responsibility for education at European Congresses, training of trainers in epilepsy (now 45 trainers from 27 countries), adoption of a curriculum to become a certified European epileptologist, establishment of a multinational educational network, and a "Website Academy" as a forum for discussion. In the first two years of its activity, the Academy received financial support from the ILAE (money generated by the European Congresses) but has since been self-supporting, with income from membership fees, participation fees at courses, and moderate support from pharmaceutical companies. The interest in the Academy is high, especially in those parts of Europe where epileptology was until recently not well developed. This should encourage other regional ILAE commissions to implement similar strategies, even if different regional structures will require variable regional approaches. Thus, in regions with long distances, it may be useful to include methods of distant learning and other specific features. PMID- 15816982 TI - Overview and perspectives of employment in people with epilepsy. AB - Even though it is now the viewpoint of the majority of professionals working in epilepsy care that most people with epilepsy should and can perform on the labor market as does anybody else, research tells a different story. Most figures concerning employment rates of people with epilepsy indicate that they do not perform as well on the labor market as others do. Although both research figures and research groups vary, generally unemployment rates are higher for people with epilepsy than for the general population. Early studies showed that the situation for people with epilepsy was rather grim. Later studies showed similar outcomes. Unemployment rates vary between groups and countries. Research shows that being employed is an important ingredient of the quality of life of people with epilepsy. The World Health Organization also recognizes the importance of employment as a part of social health, and therefore, improving the quality of life. It is important to know the perspectives on the labor market for people with epilepsy and what the possible problems are. I describe a Dutch research project and give an overview of the findings concerning the employment and consequent employability of people with epilepsy and questions pertaining to employment and epilepsy. Possible interventions [i.e., public education and employment programs for people with epilepsy with the aim to improve the (re)integration of people with epilepsy into the labor market, thus improving the quality of life of (potential) employees with epilepsy], are described extensively. PMID- 15816983 TI - Vocational assessment and intervention for people with epilepsy. AB - Employment restrictions have been experienced by many people with epilepsy. In many cases, the restrictions are unjustified and based on stigma or a stereotypical image of the person with epilepsy. Unjustifiable restrictions are a form of discrimination and lead to unemployment and underemployment. Unfortunately, much of the research in this area has been difficult to interpret because of differences in the definition of "people with epilepsy" and differences in the definition of "employment restrictions or problems." I report on an attempt to develop a classification structure and examine some survey results collected by the IBE Employment Commission from professionals and people with epilepsy concerning the sources of employment restrictions and possible methods to overcome these restrictions. PMID- 15816984 TI - Intercountry exchange and support within the region. AB - The foundation of the Asian and Oceanian Epilepsy Organization dates back to the 25th Annual Meeting of the Japan Epilepsy Society held in Shizuoka, Japan, in 1991, to which roughly a dozen epilepsy specialists were invited from neighboring countries. The objectives of this volunteers' group were to strengthen kinship among medical professionals involved in epilepsy in the region. Since then, visitors to the National Epilepsy Center in Shizuoka, some for weeks and some for months or years, joined us to establish in-service training. In the 1990s, epilepsy societies or research groups were founded in individual countries. On the occasion of meetings, mutual exchange of faculty members has become more and more frequent. This is a grass-roots movement to nurture colleagues interested in epileptology. In 1995, the International League Against Epilepsy officially launched the Commission on Asian and Oceanian Affairs to stimulate epileptology and enhance chapter formation. The Asian and Oceanian Congress was inaugulated in Seoul, Korea, in 1996, followed by the 2nd congress in Taipei, Taiwan, in 1998; the 3rd, in New Delhi, India, in 2000; and the 4th in Nagano, Japan, in 2002. The Commission also organized Epilepsy Teaching Courses in Vietnam, Indonesia, Myanmar, Mongolia, and Nepal to reinforce infrastructures of epilepsy management. The Asian and Oceanian Epilepsy Academy, now being organized, will bring about more active programs in exchange and support for medical professionals in the region. PMID- 15816985 TI - "Out of the shadows": a partnership that brings progress! PMID- 15816986 TI - Development of a SEARO report on country needs and resources for the control of epilepsy. PMID- 15816987 TI - The Western pacific region and the global campaign against epilepsy. PMID- 15816988 TI - Validation of diagnostic criteria for major seizures at the community level and in a hospital setting. PMID- 15816991 TI - Tamm-Horsfall protein: a multilayered defence molecule against urinary tract infection. AB - Urinary tract infection (UTI) is the most common nonepidemic bacterial infection in humans, representing a constant danger for the host. Both innate and adaptive components of the immune system as well as stromal cells including bladder epithelium are involved in the prevention and clearance of UTI. However, the particular properties of the urogenital tract, which does not comprise typical physical barriers like a mucus or ciliated epithelium, necessitate soluble mediators with potent immunomodulatory capabilities. One candidate molecule capable of both mediating direct antimicrobial activity and alerting immune cells is the evolutionary conserved Tamm-Horsfall protein (THP). Tamm-Horsfall protein is exclusively produced by the kidney in the distal loop of Henle; however, its definite physiological function remains elusive. Mounting evidence indicates that beyond a mere direct antimicrobial activity, THP exerts potent immunoregulatory activity. Furthermore, the genetic ablation of the THP gene leads to severe infection and lethal pyelonephritis in an experimental model of UTI. Recent data are provided demonstrating that THP links the innate immune response with specific THP-directed cell-mediated immunity. In light of these novel findings we discuss the particular role of THP as a specialized defence molecule. We propose an integrated model of protective mechanisms against UTI where THP acts by two principle nonmutually exclusive mechanisms involving the capture of potentially dangerous microbes and the ability of this peculiar glycoprotein to induce robust protective immune responses against uropathogenic bacteria. PMID- 15816992 TI - Pathogenesis, detection and treatment of Achilles tendon xanthomas. AB - Tendon xanthomatosis often accompanies familial hypercholesterolaemia, but it can also occur in other pathologic states. Achilles tendons are the most common sites of tendon xanthomas. Low-density lipoprotein (LDL) derived from the circulation accumulates into tendons. The next steps leading to the formation of Achilles tendon xanthomas (ATX) are the transformation of LDL into oxidized LDL (oxLDL) and the active uptake of oxLDL by macrophages within the tendons. Although physical examination may reveal Achilles tendon xanthomas (ATX), there are several imaging methods for their detection. It is worth mentioning that ultrasonography is the method of choice in everyday clinical practice. Although several treatments for Achilles tendon xanthomas (ATX) have been proposed (LDL apheresis, statins, etc.), they target mostly in the treatment of the basic metabolic disorder of lipid metabolism, which is the main cause of these lesions. In this review we describe the formation, detection, differential diagnosis and treatment of ATX as well as the relationship between tendon xanthomas and atheroma. PMID- 15816993 TI - Increased glomerular and extracellular malondialdehyde levels in patients and rats with focal segmental glomerulosclerosis. AB - BACKGROUND: Evidence suggests an increase in oxidative stress in patients with chronic kidney disease, as glomerulosclerosis is the prerequisite for chronic kidney disease; whether the oxidative stress already exists early on is not known. MATERIALS AND METHODS: In this study we measured the plasma and urinary levels of malondialdehyde (MDA), the end product of lipid peroxidation, and assessed the immunoreactivity of MDA and superoxide dismutase (SOD) in glomeruli of patients and rats with primary focal segmental glomerulosclerosis (FSGS), and compared our findings with those of minimal change disease (MCD) and normal controls (NC). RESULTS: Our results showed that plasma MDA level was significantly increased in patients with FSGS compared with both patients with MCD and normal controls. The urinary MDA level was also significantly increased and was significantly correlated with plasma MDA level in patients with FSGS. The immunostaining for glomerular MDA and SOD was significantly higher in the patients with FSGS than in either the patients with MCD or NC, and was also significantly higher in rats with puromycin aminonucleoside (PAN)-induced FSGS than in rats with MCD. Glomerular MDA level was significantly correlated with the degree of glomerulosclerosis in the patients with FSGS. CONCLUSIONS: Our data suggest that oxidative stress occurs early on before the onset of renal failure, and may play an important role in the pathogenesis of glomerulosclerosis. PMID- 15816994 TI - Effects of simvastatin on blood lipids, vitamin E, coenzyme Q10 levels and left ventricular function in humans. AB - BACKGROUND: As statin therapy has been reported to reduce antioxidants such as vitamin E and coenzyme Q10 and there are indications that this reduction may cause impairment of left ventricular function (LVF), we studied the influence of simvastatin on LVF and serum vitamin E and coenzyme Q10 levels in humans. MATERIAL AND METHODS: We assessed the effect of simvastatin on left ventricular function and coenzyme Q10 levels in 21 (11 male, 10 female) hypercholesterolaemic subjects (mean age = 56 years) with normal LVF, over a period of 6 months. Subjects were re-tested after a 1-month wash-out period (7 months). Echocardiography was performed on all subjects before commencement of simvastatin (20 mg day(-1)), and at 1, 3, 6 and 7 months after initiation of treatment. Fasting blood samples were also collected at these intervals to assess lipids, apoproteins, vitamin E and coenzyme Q10. RESULTS: Serum lipids showed the expected reductions. Plasma vitamin E and coenzyme Q10 levels were reduced by 17 +/- 4% (P < 0.01) and 12 +/- 4% (P < 0.03) at 6 months. However, the coenzyme Q10/LDL-cholesterol ratio and vitamin E/LDL-cholesterol ratio increased significantly. Left ventricular ejection fraction (EF) decreased transiently after 1 month, while no significant change was observed at 3 and 6 months. Other markers of left ventricular function did not change significantly at any time point. CONCLUSION: Despite reduced plasma vitamin E and coenzyme Q10, 20 mg of simvastatin therapy is associated with a significantly increased coenzyme Q10/LDL cholesterol ratio and vitamin E/LDL-cholesterol ratio. Simvastatin treatment is not associated with impairment in left ventricular systolic or diastolic function in hypercholesterolaemic subjects after 6 months of treatment. PMID- 15816995 TI - Dichotomous responses of inter and postprandial hyperglycaemia to short-term calorie restriction in patients with type 2 diabetes. AB - BACKGROUND: In type 2 diabetes the effects of short-term calorie restrictions on glycaemia are usually judged on fasting plasma glucose. As fasting duration rarely exceeds 2-3 h, we determined the effects of calorie restriction over different daytime periods using a continuous glucose monitoring system (CGMS) in noninsulin-using patients with type 2 diabetes. DESIGN: Fourteen poorly controlled (mean HbA1c = 9.3%) overweight or obese patients (mean body mass index = 30.1 +/- 0.7 kg m(-2)) with type 2 diabetes were investigated twice with a CGMS, at baseline and at the end of a 18-day calorie-restricted diet (1490 kcal day(-1)). Areas under curves (AUCs) of 24-h glucose monitoring were measured and divided into postprandial and interprandial AUCs according to meal times. RESULTS: Areas under curves of 24-h glucose monitoring were significantly decreased after dieting: -37%, P = 0.0047. Both post and interprandial AUCs were significantly diminished (-26%, P = 0.0186 and -48%, P = 0.0037, respectively), but changes in interprandial AUCs were more marked than changes in postprandial AUCs (P = 0.0060). Nycthemeral peaks of glucose were observed at mid-morning times and were not significantly different before (242 +/- 15 mg dL(-1)) and after dieting (227 +/- 16 mg dL(-1)). CONCLUSIONS: Short-term calorie restriction in overweight or obese patients with type 2 diabetes resulted in dichotomous responses between interprandial and postprandial glycaemic excursions. The resistance of mid-morning glucose peaking to calorie restriction should result in additional dietary or pharmacological measures at breakfast. PMID- 15816996 TI - Does C-reactive protein identify a subclinical metabolic disease in healthy subjects? AB - BACKGROUND: Highly sensitive C-reactive protein (hs-CRP) levels are significant predictors of subsequent diabetes and metabolic syndrome (MS). Owing the strong correlations between components of the MS and obesity with hs-CRP levels, previous studies about the associations of hs-CRP with insulin resistance might have been confounded by the inclusion of overweight or dysmetabolic subjects. DESIGN: Our aim was to evaluate the associations between hs-CRP levels and fasting insulin and insulin resistance (evaluated by the Homeostasis Model Assessment: HOMA IR) in a subgroup of subjects with normal body mass index (BMI) and without any metabolic abnormalities. Out of a cohort of 1658 middle-aged subjects, representative of the local sanitary districts of the province of Asti (north-western Italy) enrolled for metabolic screening: 241 (14.5%) showed normal BMI, glucose tolerance, blood pressure and waist values and no dyslipidaemia. RESULTS: In this subgroup of subjects, those with hs-CRP levels > or = 3 mg L(-1) showed significantly higher median insulin and HOMA-IR values (respectively: 20.4 vs. 6.0 pmol L(-1), and 0.8 vs. 0.2 microU mL(-1)x mmol L(-1)). In a multiple regression model, insulin and insulin resistance remained significantly and independently related to hs-CRP levels, after adjustments for age, sex, BMI, waist, alcohol consumption, level of physical activity and smoking habits. Very few individuals within lower fasting insulin quartiles showed hs-CRP values > or = 3 mg L(-1) when compared with approximately 60% of those within the highest quartile. CONCLUSIONS: The novel finding is that a state of low-grade systemic inflammation is present in normal BMI subjects who show subclinical insulin resistance but no other metabolic abnormalities. PMID- 15816997 TI - Indomethacin-induced ileitis is associated with tensiometric, vascular and oxidative changes in the experimental rat model. AB - BACKGROUND: Indomethacin-induced enteritis is a model of inflammatory bowel disease. MATERIALS AND METHODS: To further characterize this model, rats received two injections of indomethacin (7.5 mg kg(-1)) 24 h apart and histological damage of intestinal mucosa, oxidative stress, alterations of intestinal motility and mesenteric vascular bed (MVB) reactivity were investigated after 5 days. RESULTS: The results show that indomethacin caused several histological and functional changes at the ileal level. In particular, response to carbachol as well as the nonadrenergic-noncholinergic inhibitory response to electrical field stimulation (EFS) was lower in the treated than control rats. Moreover, nitric oxide (NO) component of the inhibitory response was higher in the treated than control rats. Mesenteric vessels preparations from the treated rats showed increased noradrenaline (NA)-induced perfusion pressure, whereas relaxant responses to acetylcholine, although not significantly reduced in the treated rats, had a higher nitrergic component. This finding suggests that vascular dysfunction may contribute to chronic inflammation. Indomethacin injection also determined acute and severe oxidative stress in ileum mucosa. CONCLUSIONS: In conclusion, our study contributes to further characterize the rat model of indomethacin-induced enteritis and suggests that it is suitable for drug screening in rats, as this model can be obtained in a very short period and is simple and reproducible. PMID- 15816998 TI - Cancer immunotherapy by fusions of dendritic and tumour cells and rh-IL-12. AB - BACKGROUND: Vaccination with fusion cells (FCs) comprising dendritic cells and tumour cells as well as administration of interleukin-12 (IL-12) showed a significant therapeutic effect against established tumours in mouse experimental models. We conducted immunotherapy against various malignant tumours using the FCs and rhIL-12, and investigated the safety and efficacy of the therapy. MATERIALS AND METHODS: Patients' DCs were mixed with autologous irradiated tumour cells and treated with 50% polyethylene glycol to generate FCs. The FCs were inoculated intradermally, and then 30 ng kg(-1) of rhIL-12 was injected at the same sites 2 and 6 days later. This process was carried out as one cycle, and three of these cycles were repeated at 1-week intervals to comprise one course. After completing the course, its safety and therapeutic effects were estimated. RESULTS: The most frequently observed adverse event was fever, observed in 26% of patients in the first cycle. Decrease in white blood cell and an increase in serum ALT were observed in 28% and 25%, respectively. Three out of 12 patients with a malignant brain tumour (25%) achieved a partial response (PR), but other patients with a malignant tumour showed no regression of their tumours. Thirteen out of 16 patients with a brain tumour (81%) showed cutaneous delayed hypersensitivity responses. However, only one of 16 patients (6%) with a malignant tumour other than a brain tumour developed such responses. CONCLUSIONS: Immunotherapy using a FC vaccine and rhIL-12 induced no serious adverse reactions, and provided good therapeutic responses in some of the patients with a brain tumour. PMID- 15816999 TI - Feasible model for prevention of functional decline in older people: municipality randomized, controlled trial. AB - OBJECTIVES: To investigate the effect of an educational program for preventive healthcare professionals in routine primary care on functional ability, nursing home admissions, and mortality in older adults. DESIGN: A prospective, controlled 3-year follow-up study (1999-2001) in primary care with randomization and intervention at the municipality level and outcomes measured at the individual level in two age cohorts. SETTING: Primary care. PARTICIPANTS: Of 81 eligible municipalities in four counties, 34 agreed to participate. A total study population of 5,788 home-dwelling subjects aged 75 and 80 were asked to participate. Written consent was obtained from 4,060 persons (70.1%), of whom 2,104 were living in 17 intervention municipalities and 1,956 were living in 17 matched control municipalities. INTERVENTION: Intervention municipality visitors received ongoing education, and local general practitioners were introduced to a short geriatric assessment program early in the study period. Control municipalities visitors and general practitioners received no education. MEASUREMENTS: At the 3-year follow-up, the outcome measures of mortality and nursing home admissions were obtained from all, and the outcome measure of functional ability was obtained from 3,383 (95.6%) of 3,540 surviving participants. RESULTS: Education improved functional ability (odds ratio=1.20, 95% confidence interval (CI)=1.01-1.42, P=.04) in intervention municipality participants, notably in the 80-year-olds. There were no differences in mortality (relative risk (RR)=1.06, 95% CI=0.87-1.28, P=.59) or rates of nursing home admissions after 3 years (RR=0.74, 95% CI=0.50-1.09, P=.13). Subjects aged 80 benefited from accepting and receiving in-home assessment with regular follow ups. CONCLUSION: A brief, feasible educational program for primary care professionals helps preserve older people's functional ability. PMID- 15817000 TI - The course of functional decline in older people with persistently elevated depressive symptoms: longitudinal findings from the Cardiovascular Health Study. AB - OBJECTIVES: To examine the relationship between persistently high depressive symptoms and long-term changes in functional disability in elderly persons. DESIGN: A community-based, prospective, observational study. SETTING: Participant data from the Cardiovascular Health Study. PARTICIPANTS: From the overall sample of 5,888 subjects, three types of participants were identified for this study: (1) persistently depressed individuals, who experienced an onset of depressive symptoms that persisted over 4 years (n=119); (2) temporarily depressed individuals, who experienced an onset of depressive symptoms that resolved over time (n=259); and (3) nondepressed individuals, with persistently low depressive symptoms throughout the follow-up period who were matched on baseline activity of daily living (ADL) scores, sex, and age to the previous two groups combined (n=378). MEASUREMENTS: Four consecutive years of data were assessed: validated measures of depression (10-item CES-D), functional disability (10-item ADL/instrumental ADL measure), physical performance, medical illness, and cognition. RESULTS: The persistently depressed group showed a greater linear increase in functional disability ratings than the temporarily depressed and nondepressed groups. This association between persistent depression and functional disability was robust even when controlling for baseline demographic and clinical/performance measures, including cognition. The persistently depressed group had an adjusted odds ratio (OR) of 5.27 (95% confidence interval (CI) 3.03-9.16) for increased functional disability compared with the nondepressed group over 3 years of follow-up, whereas the temporarily depressed group had an adjusted OR of 2.39 (95% CI=1.55-3.69) compared with the nondepressed group. CONCLUSION: Persistently elevated depressive symptoms in elderly persons are associated with a steep trajectory of worsening functional disability, generating the hypothesis that treatments for late-life depression need to be assessed on their efficacy in maintaining long-term functional status as well as remission of depressive symptoms. These results also demonstrate the need for studies to differentiate between persistent and temporary depressive symptoms when examining their relationship to disability. PMID- 15817001 TI - The effect of a task-oriented walking intervention on improving balance self efficacy poststroke: a randomized, controlled trial. AB - OBJECTIVES: To evaluate the efficacy of a task-oriented walking intervention in improving balance self-efficacy in persons with stroke and to determine whether effects were task-specific, influenced by baseline level of self-efficacy and associated with changes in walking and balance capacity. DESIGN: Secondary analysis of a two-center, observer-blinded, randomized, controlled trial. SETTING: General community. PARTICIPANTS: Ninety-one individuals with a residual walking deficit within 1 year of a first or recurrent stroke. INTERVENTION: Task oriented interventions targeting walking or upper extremity (UE) function were provided three times a week for 6 weeks. MEASUREMENTS: Activities-specific Balance Confidence Scale, Six-Minute Walk Test, 5-m walk, Berg Balance Scale, and Timed "Up and Go" administered at baseline and postintervention. RESULTS: The walking intervention was associated with a significantly greater average proportional change in balance self-efficacy than the UE intervention. Treatment effects were largest in persons with low self-efficacy at baseline and for activities relating to tasks practiced. In the walking group, change in balance self-efficacy correlated with change in functional walking capacity (correlation coefficient=0.45, 95% confidence interval=0.16-0.68). Results of multivariable modeling suggested effect modification by the baseline level of depressive symptoms and a prognostic influence of age, sex, comorbidity, time poststroke, and functional mobility on change in self-efficacy. CONCLUSION: Task-oriented walking retraining enhances balance self-efficacy in community-dwelling individuals with chronic stroke. Benefits may be partially the result of improvement in walking capacity. The influence of baseline level of self efficacy, depressive symptoms, and prognostic variables on treatment effects are of clinical importance and must be verified in future studies. PMID- 15817002 TI - High prevalence of Chlamydia pneumoniae antibodies and increased high-sensitive C reactive protein in patients with vascular dementia. AB - OBJECTIVES: To determine the relationships between Chlamydia pneumoniae infection, carotid atherosclerosis, and dyslipidemia in patients with vascular dementia (VaD) and Alzheimer's disease (AD). DESIGN: Case control study. SETTING: Showa University Karasuyama Hospital, Tokyo, Japan. PARTICIPANTS: One hundred twenty-four elderly subjects: 31 with VaD, 61 with AD, and 32 age-matched controls without dementia. MEASUREMENTS: Presence of antibodies to C. pneumoniae (immunoglobulin G (IgG) and IgA), the serum concentrations of high-sensitive C reactive protein (hs-CRP) and atherogenic lipoproteins, and the carotid artery intima-media thickness (IMT) and plaques were determined. RESULTS: Age; body mass index; systolic and diastolic blood pressures; and fasting plasma glucose, hemoglobin A(1c), high-density lipoprotein cholesterol, and apolipoprotein A-I, B, and E concentrations did not differ significantly between the three groups, but the mean IMT and frequency of atherosclerotic plaques in the carotid arteries, as well as the serum concentrations of low-density lipoprotein cholesterol (LDL-C), lipoprotein(a), and lipid peroxides were significantly greater in VaD patients than in AD patients or nondemented controls. Hs-CRP concentrations and prevalence of C. pneumoniae IgG and IgA antibodies also were significantly higher in VaD patients than in AD patients and nondemented controls. Multiple logistic regression analysis revealed that carotid IMT and plaques, LDL-C, lipid peroxides, hs-CRP, and IgG and IgA C. pneumoniae seropositivity were independent risk factors for VaD. CONCLUSION: These results suggest that carotid atherosclerosis, atherogenic lipoproteins, and C. pneumoniae infection (as documented by the IgG and IgA seropositivity together with increased hs-CRP) may be VaD risk factors. PMID- 15817003 TI - Outcomes of infection in nursing home residents with and without early hospital transfer. AB - OBJECTIVES: To compare outcomes of infection in nursing home residents with and without early hospital transfer. DESIGN: Observational cohort study. SETTING: Fifty-nine nursing homes in Maryland. PARTICIPANTS: Two thousand one hundred fifty-three individuals admitted to nursing homes between 1992 and 1995. MEASUREMENTS: Incident infection was recorded when a new infectious diagnosis was documented in the medical record or nonprophylactic antibiotic therapy was prescribed. Early hospital transfer was defined as transfer to the emergency department or admission to the hospital within 3 days of infection onset. Infection, resident, and facility characteristics were entered into a multivariate model to create a propensity score for early hospital transfer. Association between early hospital transfer and outcomes of infection, namely pressure ulcers and death between Days 4 and 34 after infection onset, were examined, controlling for propensity score. RESULTS: Four thousand nine hundred ninety infections occurred in 1,301 residents. Genitourinary (28%), skin (19%), upper respiratory (13%), and lower respiratory (12%) were the most common types. Three hundred seventy-five episodes in which residents survived 3 days (7.6%) resulted in early hospital transfer. In multivariate regression, individuals with early hospital transfer had higher mortality (odds ratio (OR) 1.44, 95% confidence interval (CI)=1.04-1.99) and, in 1-month survivors, a greater occurrence of pressure ulcers (OR 1.61, 95% CI=1.17-2.20) than those without, after adjusting for propensity score. CONCLUSION: Using observational data and propensity score methods, outcomes were worse in nursing home residents transferred to the hospital within 3 days of infection onset than in those who remained in the nursing home. PMID- 15817004 TI - The association between specific nurse case management interventions and elder health. AB - OBJECTIVES: To describe the association between specific nursing interventions performed in the context of nurse case management and older people's quality of life and functional ability. DESIGN: Longitudinal. SETTING: Nurse case management through a university hospital and two community health centers. PARTICIPANTS: One hundred seventy-five community-dwelling frail older persons (> or =70 and at risk for repeated hospitalizations). MEASUREMENTS: Specific groups of nursing interventions provided in the context of nurse case management over a 10-month period--coping assistance, lifespan care, risk management, and physical comfort promotion--were focused on. These interventions were recorded using a standardized nursing language. Outcomes were measured using telephone and home interview and medical record review using the 36-item Short Form and the Older American Resources and Services Multidimensional Functional Assessment Questionnaire. RESULTS: Older people receiving coping assistance interventions demonstrated an increase in instrumental activity of daily living functioning although they had lower general health, role-emotional, and mental health scores. CONCLUSION: Coping assistance is one nursing intervention of several provided in the context of nurse case management that is independently associated with improving the functional status of frail older persons even in the presence of declining health normally associated with aging over several months. Examining the relationships between specific nursing activities and health outcomes of frail older persons may be useful in furthering understanding of the results of randomized trials of nurse case management in this population. PMID- 15817005 TI - Purchasing or providing nursing home care: can quality of care data provide guidance. AB - OBJECTIVES: To examine whether quality of care differed for veterans in Department of Veterans Affairs (VA) nursing homes and those on contract in community nursing homes, and whether the VA was contracting with nursing homes providing better quality of care than other nursing homes. DESIGN: Observational study using administrative databases from 1997 to 1999. SETTING: Ten VA and 650 community nursing homes in New York state. PARTICIPANTS: Four thousand seven hundred sixty-three veteran and 195,438 nonveteran residents of these nursing homes. MEASUREMENTS: Risk-adjusted rates of pressure ulcer development, functional decline, behavioral decline, and mortality. RESULTS: Veterans in VA nursing homes were significantly (P< .05) less likely to develop a pressure ulcer (odds ratio (OR)=0.63) but more likely to experience functional decline (OR=1.6) than veterans in community nursing homes. Residents of community nursing homes with VA contracts were significantly (P< .05) less likely to develop a pressure ulcer (OR=0.91) but more likely to die than residents in noncontract homes. Few nursing homes were consistently among the best or worst performers on all measures; only seven of 650 nursing homes were in the top or bottom decile and 34 in the top or bottom quartile for each measure. CONCLUSION: Large purchasers and providers of nursing home care such as the VA are unlikely to find information on quality of care useful in making decisions on whether they should "make" or "buy" care. Nursing homes performing well on one quality measure may perform poorly on another, and it is difficult to identify nursing homes that are consistently among the best or worst. Other consumers may encounter similar difficulties when using data on nursing home quality. PMID- 15817006 TI - Coronary artery calcium: associations with brain magnetic resonance imaging abnormalities and cognitive status. AB - OBJECTIVES: To evaluate the association between coronary atherosclerosis and subclinical brain magnetic resonance imaging (MRI) abnormalities and between coronary atherosclerosis and abnormal cognitive function (dementia/mild cognitive impairment). DESIGN: Cross-sectional. SETTING: The Cardiovascular Health Study (CHS), an epidemiological study of risk factors for cardiovascular disease in older adults. PARTICIPANTS: Four hundred nine men and women, mean age 79, recruited from the Pittsburgh center of the CHS. MEASUREMENTS: Coronary atherosclerosis was defined according to the level of coronary artery calcification (CAC), as measured using electronic beam tomography. Subclinical brain MRI abnormalities included ventricular enlargement, white matter hyperintensities, and number of subcortical brain infarcts. Brain MRI and CAC measurements were performed between 1998 and 2000 at the Pittsburgh center of the CHS. Prevalence of brain MRI abnormalities and abnormal cognitive status were examined across quartiles of the CAC score, before and after controlling for age. Multivariate logistic regression models were used to assess whether CAC level was associated with abnormalities of brain MRI or abnormal cognitive status. RESULTS: Older adults with high CAC scores were more likely to have more-severe brain MRI abnormalities, including subcortical infarction and high white matter hyperintensities. The associations between CAC and ventricular enlargement showed a similar but not significant trend. The presence of any of the MRI abnormalities attenuated the association between CAC and abnormal cognitive status. CONCLUSION: Older adults with higher levels of CAC were more likely to have more-severe brain MRI abnormalities and abnormal cognitive status. PMID- 15817007 TI - Subjective cognitive complaints and cognitive decline: consequence or predictor? The epidemiology of vascular aging study. AB - OBJECTIVES: To explore whether more cognitive complaints are associated with previous or future cognitive decline. DESIGN: Longitudinal; epidemiology vascular aging study. SETTING: Community in Nantes, France. PARTICIPANTS: Seven hundred thirty-three subjects, aged 59 to 71. MEASUREMENTS: Subjective cognitive complaints were recorded at 4-year follow-up examination in a prospective study of people aged 59 to 71 at study entry. Participants' cognitive performances were assessed repeatedly at each wave (baseline, 4 years, and 6 years) of the study using a series of neuropsychological tests including the Mini-Mental State Examination. Depressive symptoms were evaluated using the Center for Epidemiological Studies Depression Scale. Subjects also had a cerebral magnetic resonance imaging scan at 4-year follow-up to evaluate presence and severity of white matter hyperintensities (WMHs). RESULTS: Subjects with more cognitive complaints had greater cognitive decline. This significant relationship persisted after adjusting for potential confounders, including depressive symptoms. Multivariate analysis also showed that, in subjects without measured cognitive decline between study entry and 4-year follow-up, those with more cognitive complaints at 4-year follow-up had significantly greater measured cognitive decline during the subsequent 2 years. In the presence of severe WMH, more cognitive complaints were an even stronger predictor of future cognitive decline. CONCLUSION: Cognitive complaints reflect measured cognitive decline, but they also predict cognitive decline at an earlier stage than objective tests that are not able to detect cognitive deficits. They need to be taken into account in clinical practice. PMID- 15817008 TI - A multifactorial intervention program reduces the duration of delirium, length of hospitalization, and mortality in delirious patients. AB - OBJECTIVES: To investigate whether an education program and a reorganization of nursing and medical care improved the outcome for older delirious patients. DESIGN: Prospective intervention study. SETTING: Department of General Internal Medicine, Sundsvall Hospital, Sweden. PARTICIPANTS: Four hundred patients, aged 70 and older, consecutively admitted to an intervention or a control ward. INTERVENTION: The intervention consisted of staff education focusing on the assessment, prevention, and treatment of delirium and on caregiver-patient interaction. Reorganization from a task-allocation care system to a patient allocation system with individualized care. MEASUREMENTS: The patients were assessed using the Organic Brain Syndrome Scale and the Mini-Mental State Examination on Days 1, 3, and 7 after admission. Delirium was diagnosed according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria. RESULTS: Delirium was equally common on the day of admission at the two wards, but fewer patients remained delirious on Day 7 on the intervention ward (n=19/63, 30.2% vs 37/62, 59.7%, P=.001). The mean length of hospital stay+/ standard deviation was significantly lower on the intervention ward then on the control ward (9.4+/-8.2 vs 13.4+/-12.3 days, P<.001) especially for the delirious patients (10.8+/-8.3 vs 20.5+/-17.2 days, P<.001). Two delirious patients in the intervention ward and nine in the control ward died during hospitalization (P=.03). CONCLUSION: This study shows that a multifactorial intervention program reduces the duration of delirium, length of hospital stay, and mortality in delirious patients. PMID- 15817009 TI - Prevalence and correlates of fecal incontinence in community-dwelling older adults. AB - OBJECTIVES: To determine prevalence and correlates of fecal incontinence in older community-dwelling adults. DESIGN: A cross-sectional, population-based survey. SETTING: Participants interviewed at home in three rural and two urban counties in Alabama from 1999 to 2001. PARTICIPANTS: The University of Alabama at Birmingham Study of Aging enlisted 1,000 participants from the state Medicare beneficiary lists. The sample was selected to include 25% black men, 25% white men, 25% black women, and 25% white women. MEASUREMENTS: The survey included sociodemographic information, medical conditions, health behaviors, life-space assessment (mobility), and self-reported health status. Fecal incontinence was defined as an affirmative response to the question "In the past year, have you had any loss of control of your bowels, even a small amount that stained the underwear?" Severity was classified as mild if reported less than once a month and moderate to severe if reported once a month or greater. RESULTS: The prevalence of fecal incontinence in the sample was 12.0% (12.4% in men, 11.6% in women; P=.33). Mean age+/-standard deviation was 75.3+/-6.7 and ranged from 65 to 106. In a forward stepwise logistic regression analysis, the following factors were significantly associated with the presence of fecal incontinence in women: chronic diarrhea (odds ratio (OR)=4.55, 95% confidence interval (CI)=2.03-10.20), urinary incontinence (OR=2.65, 95% CI=1.34-5.25), hysterectomy with ovary removal (OR=1.93, 95% CI=1.06-3.54), poor self-perceived health status (OR=1.88, 95% CI=1.01-3.50), and higher Charlson comorbidity score (OR=1.29, 95% CI=1.07-1.55). The following factors were significantly associated with fecal incontinence in men: chronic diarrhea (OR=6.08, 95% CI=2.29-16.16), swelling in the feet and legs (OR=3.49, 95% CI=1.80-6.76), transient ischemic attack/ministroke (OR=3.11, 95% CI=1.30-7.41), Geriatric Depression Scale score greater than 5 (OR=2.83, 95% CI=1.27-6.28), living alone (OR=2.38, 95% CI=1.23-4.62), prostate disease (OR=2.29, 95% CI=1.04-5.02), and poor self-perceived health (OR=2.18, 95% CI=1.13 4.20). The following were found to be associated with increased frequency of fecal incontinence in women: chronic diarrhea (OR=6.39, 95% CI=2.25-18.14), poor self-perceived health (OR=5.37, 95% CI=1.75-16.55), and urinary incontinence (OR=4.96, 95% CI=1.41-17.43). In men, chronic diarrhea (OR=5.38, 95% CI=1.77 16.30), poor self-perceived health (OR=3.91, 95% CI=1.39-11.02), lower extremity swelling (OR=2.86, 95% CI=1.20-6.81), and decreased assisted life-space mobility (OR=0.73, 95% CI=0.49-0.80) were associated with more frequent fecal incontinence. CONCLUSION: In community-dwelling older adults, fecal incontinence is a common condition associated with chronic diarrhea, multiple health problems, and poor self-perceived health. Fecal incontinence should be included in the review of systems for older patients. PMID- 15817010 TI - Sex steroid level, androgen receptor polymorphism, and depressive symptoms in healthy elderly men. AB - OBJECTIVES: To determine the prevalence of depression in a cohort of elderly men as assessed using a 30-item Geriatric Depression Scale (GDS) score and to describe the association between this score and sex steroids, androgen receptor (AR) polymorphism, and general health status. DESIGN: Observational study on the relationship between sex steroid status and health-related parameters. SETTING: Community-based. PARTICIPANTS: Ambulatory men (n=236 in 1997, n=192 in 2000) aged 70 and older at inclusion in 1996, interviewed in 1997 and 2000. MEASUREMENTS: Serum levels of testosterone, estradiol, sex hormone binding globulin (SHBG), dehydroepiandrosterone-sulfate (DHEAS), cortisol, and the AR gene cytosine, adenine, guanine (CAG)-repeat length polymorphism were determined. Free testosterone and free estradiol were calculated. Questionnaires included GDS, 36 item Short Form, and Rapid Disability Rating Scale-2. RESULTS: Median age was 75.3 years (interquartile range=73.5-78.5). A GDS score of 11 or greater was found in 30 (12.7%) men. Age and GDS score were significantly interrelated (P<.01), as were all health-assessment scores. GDS scores were not related to (free) testosterone or AR polymorphism in 1997 or 2000. In 1997 only (n=236), higher GDS scores were related to higher estradiol, free estradiol, and DHEAS levels. CONCLUSION: The data did not support a role for testosterone in depression in elderly community-based men as assessed using the GDS. PMID- 15817011 TI - Comparison of brain natriuretic peptide and probrain natriuretic peptide in the diagnosis of cardiogenic pulmonary edema in patients aged 65 and older. AB - OBJECTIVES: Differentiating cardiogenic pulmonary edema (CPE) from respiratory causes of dyspnea is difficult in elderly patients. The aim of this study was to compare the usefulness of B-type natriuretic peptide (BNP) and amino-terminal fragment BNP (proBNP), to diagnose CPE in patients aged 65 and older. DESIGN: Prospective study. SETTING: Medical emergency department of a 2,000-bed urban teaching hospital. PARTICIPANTS: Patients aged 65 and older presenting with acute dyspnea and a respiratory rate of 25 breaths/min or greater, a partial pressure of oxygen of 70 mmHg or less, or an oxygen saturation of 92% or less were included. MEASUREMENTS: Rapid BNP and proBNP assays, performed blind at admission, were compared with the final diagnosis (CPE or no CPE) as defined by an expert team. RESULTS: Two hundred two patients (mean age+/-standard deviation 80+/-9) were included; 88 (44%) had CPE. There was a strong correlation between proBNP and BNP values (correlation coefficient=0.91, P<.001). The median BNP and proBNP were higher in the group of patients with CPE (377 vs 74 pg/mL, P<.001, and 3,851 vs 495 pg/mL, P<.001, respectively). The best threshold values of BNP and proBNP were 250 pg/mL and 1,500 pg/mL, respectively. The area under the receiver operating characteristic curve was greater with BNP than with proBNP (0.85 vs 0.80, P<.05). BNP assay was more accurate in diagnosis than the emergency physician, whereas proBNP was not. Higher values of BNP and proBNP were associated with greater in-hospital mortality. CONCLUSION: BNP assay is a more useful diagnostic indicator for CPE than proBNP in patients aged 65 and older. PMID- 15817012 TI - Subclinical brain magnetic resonance imaging abnormalities predict physical functional decline in high-functioning older adults. AB - OBJECTIVES: To determine whether severity of subclinical brain magnetic resonance imaging (MRI) abnormalities predicts incident self-reported physical impairment or rate of decline in motor performance. DESIGN: Longitudinal analysis, average follow-up time: 4.0 years. SETTING: Cardiovascular Health Study (CHS). PARTICIPANTS: CHS participants with modified Mini-Mental State Examination (3MS) score of 80 or greater, no self-reported disability, no history of stroke, and at least one assessment of mobility (n=2,450, mean age=74.4). MEASUREMENTS: Brain MRI abnormalities (ventricular enlargement, white matter hyperintensities, subcortical and basal ganglia small brain infarcts), self-reported physical impairment (difficulty walking half a mile or with one or more activities of daily living), and motor performance (gait speed, timed chair stand). RESULTS: After adjusting for demographics, cardiovascular risk factors, and diseases, risk of incident self-reported physical impairment was 35% greater for those with severe ventricular enlargement than for those with minimal ventricular enlargement, 22% greater for those with moderate white matter hyperintensities than for those with minimal white matter hyperintensities, and 26% greater for participants with at least one brain infarct than for those with no infarcts. Those with moderate to severe brain abnormalities experienced faster gait speed decline (0.02 m/s per year) than those with no MRI abnormalities (0.01 m/s per year). Further adjustment for incident stroke, incident dementia, and 3MS score did not substantially attenuate hazard ratios for incident self-reported physical impairment or coefficients for decline in gait speed. CONCLUSION: Subclinical structural brain abnormalities in high-functioning older adults can increase the risk of developing physical disabilities and declining in motor performance. PMID- 15817013 TI - People aged over 75 in atrial fibrillation on warfarin: the rate of major hemorrhage and stroke in more than 500 patient-years of follow-up. AB - OBJECTIVES: To determine the incidence of major hemorrhage and stroke in people aged 76 and older with atrial fibrillation on adjusted-dose warfarin who had been recently been admitted to hospital. DESIGN: A retrospective observational cohort study. SETTING: A major healthcare network involving four tertiary hospitals. PARTICIPANTS: Two hundred thirty-five patients aged 76 and older admitted to a major healthcare network between July 1, 2001, and June 30, 2002, with atrial fibrillation on warfarin were enrolled. MEASUREMENTS: Information regarding major bleeding episodes, strokes, and warfarin use was obtained from patients, relatives, primary physicians, and medical records. RESULTS: Two hundred twenty eight patients (42% men) with a mean age of 81.1 (range 76-94) were included in the analysis. Total follow-up on warfarin was 530 years (mean 28 months). There were 53 major hemorrhages, for an annual rate of 10.0%, including 24 (45.3%) life threatening and five (9.4%) fatal bleeds. The annual stroke rate after initiation of warfarin was 2.6%. CONCLUSION: The rate of major hemorrhage was high in this old, frail group, but excluding fatalities, resulted in no long-term sequelae, and the stroke rate on warfarin was low, demonstrating how effective warfarin treatment is. PMID- 15817014 TI - Comorbidity and 1-year mortality risks in nursing home residents. AB - OBJECTIVES: To investigate the effect of chronic diseases and disease combinations on 1-year mortality in nursing home residents. DESIGN: Retrospective cohort study using electronically submitted Minimum Data Set (MDS) information and Missouri death certificate data. SETTING: Five hundred twenty-two nursing homes in Missouri. PARTICIPANTS: Forty-three thousand five hundred ten nursing home residents with a full MDS assessment in 1999. MEASUREMENTS: Information about chronic diseases, age, sex, and performance in activities of daily living (ADLs) available from the first full MDS 2.0 assessment in 1999; death within 1 year after the first full MDS-assessment in 1999. RESULTS: After adjustment for age and sex, eight variables were predictive for 1-year mortality: seven chronic diseases (dementia, cancer, heart failure, renal failure, emphysema/chronic obstructive pulmonary disease, diabetes mellitus, and anemia) and an interaction variable containing age and cancer. Adding terms for disease combinations (e.g., diabetes mellitus and heart failure) did not enhance survival prediction. When there was also adjustment for ADL performance as measured using the MDS-ADL Short Form, dementia and anemia were not included, because they had no prognostic value above that of the other variables. CONCLUSION: Several chronic diseases were associated with 1-year mortality in the institutionalized elderly after adjustment for ADL performance, age, and sex. Evidence of a synergistic effect of disease combinations on mortality is lacking. PMID- 15817015 TI - Determinants of self-management strategies to reduce out-of-pocket prescription medication expense in homebound older people. AB - OBJECTIVES: To determine the extent to which homebound older people adopt strategies to reduce out-of-pocket prescription medication cost and the factors associated with level of cost-related medication management. DESIGN: Cross sectional study. SETTING: Home-delivered meals programs in four North Carolina counties. PARTICIPANTS: Random sample of 222 home-delivered meal recipients aged 60 and older. MEASUREMENTS: The use of six different management strategies to reduce medication expenses was reported at the in-home assessment. Associations between level of cost-related medication management and sample characteristics, drug coverage, behaviors to cope with out-of-pocket medication expense, and payment difficulty were examined. RESULTS: Forty-five (20.3%) participants used one or more behaviors that restricted medication use; another 47 (21.2%) used one or more strategies to reduce out-of-pocket medication cost. Using medication restriction to reduce medication expense was more likely in older people who had difficulty paying for medications (odds ratio (OR)=8.2, 95% confidence interval (CI)=1.4-50.3), or used a strategy to cope with out-of-pocket expenses (choose food or medications (OR=5.1, 95% CI=1.7-15.7) or borrowed money or had another person pay for medications (OR=5.5, 95% CI=2.6-11.6)). Income, drug coverage, and medication use (prescribed and over-the-counter) increased the likelihood of having increased difficulty paying for medications. CONCLUSION: Clinicians should attempt to identify patients who are at risk for medication restriction and develop strategies for minimizing any unintended consequences of cost-related medication management behaviors. Provider-patient communication should include discussion of medication cost and appropriate medication management strategies. PMID- 15817016 TI - Dissemination of an evidence-based multicomponent fall risk-assessment and management strategy throughout a geographic area. AB - OBJECTIVES: To report on the penetration of, and identified barriers to and facilitators of, efforts to incorporate evidence-based fall risk assessment and management into clinical practice throughout a defined geographic area. DESIGN: Dissemination project. SETTING: North central Connecticut. PARTICIPANTS: Hospitals, home care agencies, primary care providers, and outpatient rehabilitation facilities. INTERVENTION: Multiple professional behavior-change strategies were used to encourage providers to incorporate evidence-based fall assessment and management into their practices. MEASUREMENTS: Penetration of dissemination efforts over 36 months; barriers and facilitators identified by provider working groups during the first 2 years of the project. RESULTS: All seven hospitals and 26 home care agencies in the area, 119 of 130 rehabilitation facilities, and 138 of 212 primary care offices participated. Most provider working groups expressed similar barriers and facilitating factors. Reported barriers specific to fall risk management included lack of awareness of fall morbidity and preventability, perceived lack of expertise and Medicare coverage, inadequate referral patterns among providers, and lack of a federal mandate for physicians. Facilitating factors specific to falls included the opportunity to market new services and to develop new networks of professional relationships across disciplines and the Medicare mandate that home care agencies focus on functional outcomes. CONCLUSION: Dissemination efforts showed notable successes as well as challenges. Although many of the barriers were general to diffusing new practices, several were specific to fall assessment and management that span disciplines and sites. Project results have implications for efforts to diffuse evidence-based practices for multifactorial geriatric conditions such as falls. PMID- 15817017 TI - Near vision impairment predicts cognitive decline: data from the Hispanic Established Populations for Epidemiologic Studies of the Elderly. AB - OBJECTIVES: To estimate the association between sensory impairment and cognitive decline in older Mexican Americans. DESIGN: A prospective cohort study. SETTING: The Hispanic Established Populations for Epidemiologic Studies of the Elderly from five southwestern states. PARTICIPANTS: The sample consisted of 2,140 noninstitutionalized Mexican Americans aged 65 and older followed from 1993/1994 until 2000/2001. MEASUREMENTS: The outcome, cognitive function decline, was assessed using the Mini-Mental State Examination blind version (MMSE-blind) at baseline and at 2, 5, and 7 years of follow-up. Other variables were near vision, distance vision, hearing, demographics (age, sex, marital status, living arrangements, and education), depressive symptoms, hypertension, diabetes mellitus, stroke, heart attack, and functional status. A general linear mixed model was used to estimate cognitive decline at follow-up. RESULTS: In a fully adjusted model, MMSE-blind scores of subjects with near vision impairment decreased 0.62 points (standard error (SE)=0.29, P=.03) over 2 years and decreased (slope of decline) 0.13 points (SE=0.07, P=.045) more per year than scores of subjects with adequate near vision. Other independent predictors of cognitive decline were baseline MMSE-blind score, age, education, marital status, depressive symptoms, and number of activity of daily living limitations. CONCLUSION: Near vision impairment, but not distance vision or hearing impairments, was associated with cognitive decline in older Mexican Americans. PMID- 15817018 TI - Identifying the activities affected by chronic nonmalignant pain in older veterans receiving primary care. AB - OBJECTIVES: To identify the specific types of activities affected by chronic pain in older persons and the extent to which older individuals modify, perform less frequently, or terminate activities because of pain. DESIGN: Cross-sectional survey. SETTING: Primary care practice at a Veterans Affairs Medical Center in New England. PARTICIPANTS: Two hundred forty-four patients (aged 65-90) with chronic nonmalignant pain. MEASUREMENTS: Open-ended questions were used to identify the activities affected by pain; participants' responses were subsequently organized into distinct categories (e.g., climbing stairs under higher-order physical activities and going out to dinner under social/recreational activities). Participants were also asked to indicate whether they had modified, performed less frequently, or terminated these activities because of pain. RESULTS: Participants had a mean age+/-standard deviation of 75.4+/-5.2, were mostly male (84%), and had an average pain intensity score of 6.2+/-1.9 on a 0- to 10-scale. Two hundred three participants (83%) reported that pain affected one or more higher-order physical activities, and the corresponding percentages for the categories of social/recreational activities, instrumental activities of daily living, and basic activities of daily living were 74%, 57%, and 3%, respectively. The proportions of participants who modified, performed less frequently, or terminated one or more activities because of pain were 71%, 69%, and 22%, respectively. CONCLUSION: Assessing the effects of chronic pain across multiple functional domains is indicated in older primary care patients, particularly higher-order physical and social/recreational activities. Inquiring about whether the activities are modified, reduced, or terminated may also help to expand understanding of pain-related disability in older persons. PMID- 15817019 TI - The Montreal Cognitive Assessment, MoCA: a brief screening tool for mild cognitive impairment. AB - OBJECTIVES: To develop a 10-minute cognitive screening tool (Montreal Cognitive Assessment, MoCA) to assist first-line physicians in detection of mild cognitive impairment (MCI), a clinical state that often progresses to dementia. DESIGN: Validation study. SETTING: A community clinic and an academic center. PARTICIPANTS: Ninety-four patients meeting MCI clinical criteria supported by psychometric measures, 93 patients with mild Alzheimer's disease (AD) (Mini Mental State Examination (MMSE) score > or =17), and 90 healthy elderly controls (NC). MEASUREMENTS: The MoCA and MMSE were administered to all participants, and sensitivity and specificity of both measures were assessed for detection of MCI and mild AD. RESULTS: Using a cutoff score 26, the MMSE had a sensitivity of 18% to detect MCI, whereas the MoCA detected 90% of MCI subjects. In the mild AD group, the MMSE had a sensitivity of 78%, whereas the MoCA detected 100%. Specificity was excellent for both MMSE and MoCA (100% and 87%, respectively). CONCLUSION: MCI as an entity is evolving and somewhat controversial. The MoCA is a brief cognitive screening tool with high sensitivity and specificity for detecting MCI as currently conceptualized in patients performing in the normal range on the MMSE. PMID- 15817020 TI - Assessing environmentally determined mobility disability: self-report versus observed community mobility. AB - OBJECTIVES: To examine the test-retest reliability and concurrent validity of a new self-report measure of mobility function by comparing it with observed mobility, self-reported activity of daily living (ADL) function, and performance based measures of gait and balance. DESIGN: Cross-sectional study involving two groups of older adults. SETTING: Community sites in Seattle, Washington, and Waterloo, Ontario, Canada. PARTICIPANTS: Fifty-four adults aged 70 and older, recruited. MEASUREMENTS: Subjects completed the Environmental Analysis of Mobility Questionnaire (EAMQ), reporting frequency of encounter and avoidance of 24 features of the physical environment, grouped into eight dimensions, on two occasions 1 week apart. Subjects were observed and videotaped during six trips into the community; frequency of encounters with environmental features within the eight dimensions was recorded. EAMQ encounter and avoidance scores were compared with observed environmental encounters, with disability in ADLs and instrumental ADLs (IADLs), and lower extremity functional measures including the Short Physical Performance Battery (SPPB) and the Berg Balance Test. RESULTS: EAMQ test-retest reliability was high for all eight dimensions (intraclass correlation coefficient range=0.81-1.0) and for summary encounter (0.98) and avoidance (0.96) scores. Observed mobility was significantly correlated (Spearman correlation = r) with EAMQ summary encounter (r=0.66) and avoidance (r=-0.58) scores. Moderate correlations were present between the EAMQ (encounter or avoidance) and observed mobility in the distance, temporal, terrain, posture, load, and density dimensions but not in the attention and ambient dimensions. EAMQ encounter/avoidance was significantly associated with ADL and IADL ability and performance on the SPPB and Berg Balance Test. CONCLUSION: Self-reported frequency of encounter and avoidance of specific environmental features appears to be a valid method for determining environmentally specific mobility disability but needs to be confirmed in a larger sample. PMID- 15817021 TI - There is hope for the future: national survey results reveal that geriatric medicine fellows are well-educated in end-of-life care. AB - OBJECTIVES: To assess the status of geriatric medicine (GM) fellows' training experiences in end-of-life care via self-report. DESIGN: Anonymous surveys completed by mail, Web access, and telephone. SETTING: U.S. accredited GM fellowship training programs. PARTICIPANTS: Two hundred ninety-six surveys were sent to graduating GM fellows in 1- and 2-year programs across the Unites States. MEASUREMENTS: Measurements assessed self-reported attitudes, quantity and quality of end-of-life care education, preparation to provide care, and perceived value of caring for dying patients. RESULTS: Response rate was 74%. Ninety-five percent or more of respondents held positive views about physicians' responsibility and ability to help dying patients. Seventy percent of fellows had completed a rotation focused on end-of-life care. Fellows who had done such rotations rated their end-of-life care education as highly as their overall geriatrics training. Fellows frequently received teaching in many end-of-life care topics, with lower rates of teaching how to say goodbye and responding to requests for assisted suicide. Overall, fellows felt well prepared to care for dying patients. Four factors independently predicted such preparedness: having had a palliative or end of-life care rotation, being female, having been taught how to say goodbye to patients, and perceiving that it is important to attending physicians that fellows learn to care for dying patients. CONCLUSION: GM fellows feel their end of-life care education is excellent and feel prepared to take care of dying patients. It is critical that geriatricians in training have access to and take advantage of palliative and end-of-life care rotations. PMID- 15817022 TI - The influence of spiritual beliefs and practices on the treatment preferences of African Americans: a review of the literature. AB - Spirituality is an important part of African-American culture and is often cited as an explanation for the more-aggressive treatment preferences of some African Americans at the end of life. This paper reviews the literature on spiritual beliefs that may influence the treatment decisions of African Americans. Medline 1966 to February 2003, Psych Info 1872 to February 2003, and CINAHL 1982 to February 2003 were searched for studies exploring spiritual beliefs that may influence the treatment preferences of African Americans. All candidate papers were examined for quality, and data were extracted on study population, design, analysis, and results to identify recurrent themes. Forty studies met inclusion criteria. Recurrent themes describing spiritual beliefs that may influence the treatment preferences of African Americans throughout the course of illness include the following: spiritual beliefs and practices are a source of comfort, coping, and support and are the most effective way to influence healing; God is responsible for physical and spiritual health; and the doctor is God's instrument. Spiritual beliefs specifically addressing treatment preferences at the end of life include: only God has power to decide life and death, there are religious prohibitions against physician-assisted death or advance directives limiting life-sustaining treatments, and divine intervention and miracles occur. For some African Americans, spiritual beliefs are important in understanding and coping with illness and may provide a framework within which treatment decisions are made. Given the growing ethnic diversity of the United States, some understanding of the complexities of culture and spirituality is essential for healthcare providers. PMID- 15817025 TI - Education, prevention, and the translation of research into practice. PMID- 15817026 TI - Improving osteoporosis management in at-risk fracture clinic patients. PMID- 15817027 TI - Long-term practice of Jaques-Dalcroze eurhythmics prevents age-related increase of gait variability under a dual task. PMID- 15817028 TI - Nonagenarians and trauma: an increasingly common combination. PMID- 15817029 TI - Tiapride may accelerate lung cancer in older people: a case report. PMID- 15817030 TI - Paroxetine and improvement of visual hallucinations in patients with dementia with lewy bodies. PMID- 15817031 TI - No positive correlation between anemia and disability in older people in Japan. PMID- 15817032 TI - Decreased serum substance P concentration in the acute phase of intracranial hemorrhage. PMID- 15817033 TI - Walking while talking: a dopamine-responsive task in early Parkinson's disease. PMID- 15817034 TI - Age and acute mountain sickness: examining the data. PMID- 15817035 TI - Prevalence of overweight and obesity in older U.S. adults: estimates from the 2003 Behavioral Risk Factor Surveillance System survey. PMID- 15817036 TI - In search of affordable prescription medications. PMID- 15817038 TI - Art, dreams and active imagination: a post-Jungian approach to transference and the image. AB - The term active imagination is sometimes applied rather uncritically to describe all forms of creative activity that take place in depth psychology. Whilst there are many forms of expression that evoke or are evoked by active imagination, they cannot automatically be classed as active imagination. In this article investigation of visualized mental imagery, dreams and art reveals three distinct forms of image-based psychological activity. Integrated and mediated within the transference and countertransference dynamic, it is proposed that the engagement in active imagination reflects and is influenced by the transference. Distinctions between sign and symbol, simple and big dreams as well as diagrammatic and embodied imagery clarify the differences. Examples from clinical practice demonstrate each mode in action within the analytic frame. PMID- 15817039 TI - An under-active or over-active internal world? An exploration of parallel dynamics within psyche and soma, and the difficulty of internal regulation, in patients with Chronic Fatigue Syndrome and Myalgic Encephalomyelitis. AB - This paper explores the dynamics brought into analytic work when there is a symmetric fusion between psyche and soma within the patient. It will consider how such a fusion may emerge from reverberations between physical constitution and a lack of maternal attunement, containment and reflective function. I will describe the work with a patient, Jane, who was diagnosed with Myalgic Encephalomyelitis (ME) during the course of her analysis. The dynamic of her physical symptoms within the analytic work, and the impact of her internal affects and internal 'objects' within the transference and countertransference, indicated a difficulty in finding an homeostatic balance resulting in overactivity and underactivity at both somatic and psychological levels. Using the clinical work with Jane this paper will also examine the interrelationship between mother-infant attachment, an inadequate internalized maternal reflective function, affect dysregulation, unconscious fusion, the lack of psyche-soma differentiation and the impact of the latter in relation to internal regulation systems, or lack of, in patients with Chronic Fatigue Syndrome (CFS) and Myalgic Encephalomyelitis (ME). I will draw on similar work carried out by Holland (1997), Simpson (1997) and Simpson et al. (1997). The paper will also employ the concept of the reflective function (Fonagy 2001; Knox 2003), and consider Matte-Blanco's (1999) concepts of generalization and unconscious symmetry in relation to the patient's internal world. I go on to consider how analysis provides a point outside the 'fusion' that can enable the 'deadlock' to be broken. PMID- 15817040 TI - The influence of complexes on implicit learning. AB - A century ago, Jung looked into the unconscious through complexes by using word association tests. Jung wrote, 'modern psychology with its investigation of complexes has opened up a psychic taboo area riddled with hopes and fears', and complexes remain an unexplored taboo area of research. In the present study, we have investigated the influence of complexes on unconscious cognitive processing, in particular on implicit learning. We have found that complexes shown to disturb conscious cognitive processing in fact enhanced the attention of the subjects and their performance on an implicit learning task. These results suggest that complexes are not just abstractions, but have various actual influences on both conscious and unconscious processing. PMID- 15817042 TI - The place of the 17th century in Jung's encounter with China. AB - After recounting several dreams and related alchemical interests of Jung's tied to the 17(th) century, a contextualizing look at select scientific and philosophical developments of that century is presented. Several precursors of the contemporary debates on the mind/body relation are noted, with special reference to the work of Antonio Damasio. This in turn leads to a reconsideration of the work of the 17(th) century polymath Gottfried Wilhelm Leibniz, which Jung read as a major precursor to his formulation of synchronicity (via Leibniz's concept of 'pre-established harmony'). Leibniz was the first philosopher to articulate the mind/body relationship in terms of supervenience, sharing an accord with those contemporary philosophers and scientists who see the mind as being an emergent property of the body-brain. Similarly, these ideas are also consistent with a reformulation of synchronicity in terms of emergence. Tracing Leibniz's interest in China reveals another set of links to Jung and to emergentism. Jung's use of Taoist concepts in developing the synchronicity principle is well known. According to scholars, Leibniz was the first major Western intellect to study the I-Ching, through the assistance of a Jesuit missionary in Beijing, Fr. Joachim Bouvet. Some details of the Leibniz-Bouvet correspondence are discussed here. Despite Helmut Wilhelm's presenting aspects of this correspondence at an Eranos conference, Jung does not appear to have integrated it into his writing on synchronicity--a possible reason for this omission is suggested. PMID- 15817043 TI - Some reflections on the influence of Chinese thought on Jung and his psychological theory. AB - Jung claimed that Richard Wilhelm, whose masterful translations of Chinese wisdom literature into a European language (German) and thence into Western consciousness have brought Chinese modes of thinking to so many, was one of the most important influences on his own life and work. The contacts between the two men, which took place from the early 1920's until Wilhelm's death in 1930, were few but intense and for Jung decisive in several ways. Wilhelm's translations of the I Ching and The Secret of the Golden Flower opened new avenues for Jung that had far-reaching consequences on his research and writing after 1930. The latter opened the door to the study of alchemy as a key to the archetypal process of individuation as rooted in the collective unconscious. 'Synchronicity' is a term that grew out of his contact with Chinese thought, in particular with the I Ching. From his contact with Chinese thought, additionally, he received confirmation of the view, independently arrived at, that adult psychological development is not linear but rather circular and spiral-like. The letters between Jung and Wilhelm illuminate the great importance Jung ascribed to Wilhelm's contribution toward bridging East and West and the potential value of Chinese philosophy for psychotherapy. PMID- 15817044 TI - Synchronicity and the I Ching: Jung, Pauli, and the Chinese woman. AB - The capacity of the human mind to discover and invent both imagistic analogies and mathematical structures to represent reality is strikingly juxtaposed in the ancient Chinese text of the I Ching. Its emphasis on containing all sorts of opposites and its plastic appeal to multi-valenced experience has kept it alive through millennia and across cultures. Jung was introduced to its Taoist wisdom by the Sinologist Richard Wilhelm. The Nobel Laureate quantum physicist Wolfgang Pauli became familiar with its philosophy and mathematics through his reading of Schopenhauer and Leibniz. In their correspondence about the nature of the unconscious and synchronicity, Pauli and Jung also exchanged their musings on Pauli's dreams of a Chinese woman, her role in his psyche and his scientific theories(1). PMID- 15817045 TI - The I Ching and the psyche-body connection. AB - Carl G. Jung's fateful meeting with Richard Wilhelm in 1929 has helped to build a bridge of depth psychological understanding between the East and the West. When Jung emerged from his 'confrontation with the unconscious', he felt validated by Wilhelm in his discovery of the healing power of medieval alchemical symbolism for the European psyche. Analytical psychology however offers a scientific, psychological understanding of Chinese wisdom as contained in the I Ching and Taoist alchemy. The Taoist alchemical tradition (also known as the Inner Elixir tradition of which 'The Secret of the Golden Flower' is a sample text) is based on the premise that psychological experience of the Tao can be achieved through mental and physiological means such as breathing and meditative techniques, gymnastics, dietary regimens such as fasting, consumption of medicinal herbs and minerals, and special sexual practices. This tradition incorporates the I Ching and traditional Chinese medicine in the alchemical opus. Taoist alchemy assumes the primacy of the physical body in the process of self-realization. The psychological and cosmic forces of the trigrams of the I Ching are stored in the internal organs of the body and are the basic material for the experience of Tao. The internal organs are the foundation of the material and subtle bodies and through cultivation, the body becomes spiritualized as the spirits are embodied. The body as a reflection of the entire cosmos becomes the residence of the gods. The realization of a new consciousness is symbolized by the hexagram Fu, meaning rebirth. The Chinese notion of Tao coincides with Jung's postulation of the unus mundus, the unity of existence which underlies the duality of psyche and matter, the psycho-physical background of existence. In this light, in the world of inner experience, East and West follow similar paths symbolically. PMID- 15817048 TI - Risk factors for coronary heart disease among inpatients who have mild intellectual disability and mental illness. AB - BACKGROUND: Coronary heart disease (CHD) is a major cause of morbidity and mortality in the UK. The aim of this study was to screen inpatients with mild or borderline intellectual disability, many of whom also have mental illness, for risk factors for CHD. METHODS: Cross sectional survey. Participants were interviewed, measured and had blood samples taken. RESULTS: Of the 53 participants, 20 (37.7%) were overweight and 18 (34.0%) obese. The mean body mass index (BMI) of those participants prescribed regular antipsychotics was higher than those who were not. Nine (20.9%) had waist circumference measurements placing them at increased risk of CHD and 21 (48.8%) were at substantially increased risk. Twenty-eight (52.8%) were current smokers. Of the 49 participants who had their blood pressure measured, 3 (6.1%) had readings above the reference range. Of the 19 participants who had random blood tests, one (5.3%) had an elevated cholesterol level. CONCLUSIONS: In this population there was a high prevalence of two risk factors for CHD (obesity and smoking), requiring ongoing monitoring and long-term measures to reduce risk. PMID- 15817049 TI - Social skills in children with intellectual disabilities with and without autism. AB - BACKGROUND: Social skills were studied in 363 children with mild intellectual disabilities (ID) and 147 with moderate ID with and without autism (age 4 through 18). The objective was to investigate the value of the Children's Social Behaviour Questionnaire (CSBQ), as a measure of subtle social skills, added to a measure of basic social skills with the Vineland Adaptive Behaviour Scales (VABS), in identifying children with ID with or without autism. METHOD: Children with mild and moderate ID, with and without autistic symptomatology were compared on basic social skills, measured with the Communication and Socialization domains of the VABS, and subtle social skills, measured with the CSBQ. RESULTS: Measuring basic social skills is not sufficient in differentiating between levels of ID. Communicative skills and subtle social skills, that concern overlooking activities or situations and fear of changes in the existing situation, seem to play a far greater role. Additionally, with respect to identifying autistic symptomatology, basic social skills do not contribute, as opposed to communicative skills and the tendency to withdraw from others. CONCLUSIONS: The results implicate that the CSBQ not only has specific value as a measure of subtle social skills to identify pervasive developmental disorders, but that the instrument also has a specific contribution to differentiating between the two levels of ID. Furthermore, our outcomes imply a slight difference between limitations in subtle social skills as mentioned by the AAMR (American Association on Mental Retardation 2002) and limitations in subtle social skills as seen in milder forms of pervasive developmental disorders. Clinical and theoretical implications will be discussed. PMID- 15817050 TI - Assessment of anger coping skills in individuals with intellectual disabilities. AB - Recent controlled studies have supported the effectiveness of anger management training for people with intellectual disabilities (IDs). This report describes an evaluation instrument designed to assess their usage of specific anger coping skills. The Profile of Anger Coping Skills (PACS) is designed for completion by a staff member or carer. Three situations are first elicited in which a client frequently displays anger. The respondent then rates each situation for the extent to which the client deploys each of eight behavioural and cognitive coping skills. In a preliminary reliability study, 20 users of a day service for people with IDs were rated independently by two staff members, with one of them completing the assessment on two separate occasions: the PACS showed good test retest reliability and lower, but still acceptable, interrater reliability. The PACS was subsequently used, in a different day service, as part of the assessment pack administered before and after a 12-week anger management group, with a parallel assessment of an untreated control group. The treated group showed substantial decreases in measures of anger, which were maintained at 6-month follow-up. Increases in PACS-rated anger coping skills were also seen in all participants in the anger management group, but not in the control group. There were differences in the extent to which different coping skills were acquired by the treated group, and there were also individual differences in the profile of specific skills acquired. It is concluded that the PACS is a reliable instrument for assessing anger coping skills, particularly when used repeatedly with the same informant. It provides information that is useful for both individual care planning and the design of future anger management programmes. PMID- 15817051 TI - Person Focused Training: a model for delivering positive behavioural supports to people with challenging behaviours. AB - BACKGROUND: Person Focused Training is introduced as a model of service delivery for people with severe challenging behaviours. It is defined as training and supporting staff to conduct functional assessments and to design and implement positive behavioural support for specific individuals with challenging behaviours. METHOD: Longitudinal outcome data are presented from 138 behaviour support plans developed by staff over a seven year period were analysed to determine reductions in frequency of challenging behaviours. Degree of behaviour change was determined across topography of behaviour, gender, age, level of disability, location of residence and role of course participant. RESULTS: Results indicate that the implementation by staff of behaviour support plans are associated with significant improvement in 77% of cases at an average follow-up of 22 months after implementation of support plans. Only location of residence was identified as related to reduction in challenging behaviours with large residential centres being associated with lower rates of behavioural improvement. CONCLUSIONS: It is argued that Person Focused training may represent an alternative to existing models of supporting individuals with challenging behaviours. The implications of front-line staff designing and implementing behaviour support plans for the organisation of services and the role of the clinical psychologist are considered. PMID- 15817052 TI - Expressive and receptive vocabulary in children with Williams and Down syndromes. AB - BACKGROUND: Williams (WS) and Down syndromes (DS) are two genetic disorders that involve intellectual disability (ID) and have been extensively studied over the past decades because of the unique linguistic profiles they exhibit. Recent investigations seek to explore the fractionation of linguistic components within the cognitive system using genetically based neurodevelopmental disorders such as WS and DS and to identify different profiles of linguistic function in these two groups of individuals. METHOD: The 'expressive vocabulary', 'receptive vocabulary', 'word opposites' and 'word definitions' subtests (Level 1) of the Test of Word Knowledge (TOWK) were used to assess lexical skills in six children with WS and five children with DS. RESULTS: Our findings indicate that the two syndromes exhibit substantial differences on linguistic tasks with individuals with WS performing at a higher level compared to those with DS and producing atypical responses in word definitions. The pattern of errors for each syndrome is qualitatively different suggesting that their underlying linguistic mechanisms are distinctive even though ID is similar. CONCLUSIONS: This study supports the differential outcome of two chromosomal disorders with similar ID. It also argues in favour of the large within group variability of the two syndromes that is not related to mental age but rather to different underlying mechanisms supporting language. These findings are discussed in the light of the current evidence concerning linguistic knowledge of neurodevelopmental and genetic disorders. PMID- 15817053 TI - A measure of subjective burden for dementia care: the Caregiving Difficulty Scale -Intellectual Disability. AB - BACKGROUND: It has been suggested in the literature on family caregiving for persons with Alzheimer's dementia (AD) that levels of objective and subjective burden among carers often predict institutionalization of the persons with AD. There is a paucity of measures to assess whether perceived burden among formal caregivers may also predict movement to more restrictive settings for persons with intellectual disabilities (ID) and AD. This study focused upon the development of a measure of subjective burden, The Caregiving Difficulty Scale - Intellectual Disability (CDS-ID) as a first step in addressing this measurement deficit. METHODS: An existing caregiver subjective burden scale, the Caregiving Hassles Scale (CHS) was adapted for use with 203 staff caregivers of persons with ID and AD. Preliminary testing of existing CHS items and proposed new items was carried out in two countries, Ireland and the USA. Confirmatory factor analysis with the existing items and exploratory factor analysis with existing and proposed new items for the scale was used to establish the content and test the psychometric properties of a revised scale, the CDS-ID. RESULTS: On the existing CHS items, staff carers appeared to experience greater subjective burden than has been reported for family caregivers. However, the psychometric properties of the CHS found with this population were poor. Factor analysis produced a revised scale, the CDS-ID with three subscales with Cronbach alphas ranging from 0.75 to 0.93 and 38 items overall. CONCLUSIONS: This new scale when used with objective burden and other scales offers an opportunity to more systematically measure the difficulties staff experience when caring for persons with ID who present with symptoms of AD. PMID- 15817054 TI - Theory of mind deficits in children with fragile X syndrome. AB - BACKGROUND: Given the consistent findings of theory of mind deficits in children with autism, it would be extremely beneficial to examine the profile of theory of mind abilities in other clinical groups such as fragile X syndrome (FXS) and Down syndrome (DS). Aim The aim of the present study was to assess whether boys with FXS are impaired in simple social situations that require them to understand their own and others' mental states - in essence: do they have a 'theory of mind'? METHOD: Well-standardized tasks of theory of mind, the location change false belief task and the appearance-reality tasks were employed to examine whether any impairment might be specific to the FXS or part of a more generalized developmental deficit. RESULTS: The results suggest that children with FXS do have impairment in theory of mind that is comparable to the deficit reported in other groups with learning disabilities such as DS. However, closer inspection of the impairment between these groups revealed qualitative differences in error types (realist vs. phenomenist), suggestive of atypical development that goes beyond general cognitive delay. CONCLUSION: The findings are discussed in terms of the teasing apart of different components of social cognition in order to identify syndrome-specific deficiencies and proficiencies. PMID- 15817055 TI - Reflection on SARS precautions in a severe intellectual disabilities hospital in Hong Kong. AB - BACKGROUND: Hong Kong went through a battle with a new respiratory disease, severe acute respiratory syndrome (SARS), from March to June 2003. All clinical settings, including rehabilitative and infirmary setting, have actively involved in fighting against the infection. The intent of this paper was to reflect on the SARS precautionary measures that had been taken in a severe intellectual disabilities hospital in Hong Kong. METHODS: A review on six SARS precautionary measures were conducted. They were assessment of risk, formulation of operational guidelines, implementation of infection control measures, education and training of staff, conducting audits and carrying out environmental improvement work. RESULTS: Patients were at risk of getting infected from carers, visitors, volunteers, and staff and patients of general hospitals. A SARS Quarantine Unit, isolation ward, was opened to isolate patients who might have had close contact with SARS patients during a stay in a general hospital or when they returned from home leave. Undoubtedly, both staff and relatives participated in preventing the patients from being infected. No day leave and home leave was reported and the number of hospitalization in general hospital was decreased during the critical period. Three infection control audits were conducted and improvement work was carried out subsequently. CONCLUSION: The practice of grouping within a standard isolation room is recommended to continue in the future. Moreover, intensive infection control training for all staff is of highest importance to safeguard the health of both staff and patient. PMID- 15817057 TI - Comments on the shortcomings of predicting the precision of Cavalieri volume estimates based upon assumed measurement functions. AB - It is well known that the estimation of an object's volume by means of serial cross-sections, the so-called Cavalieri method, yields an unbiased estimate. But by itself it provides no means by which to estimate how precise this estimate is unless the shape of the volume is fully known beforehand. This knowledge can only be partially determined from the serial section information that is collected. Methods have been developed that claim to surmount this difficulty by using the serial section data to create a mathematical model of the volume's shape properties. The model then is used to estimate (predict) the precision of the volume estimate (its CE) from the single set of data available. Unfortunately, the theory underlying the model is flawed and so the model itself amounts to no more than an unsubstantiated guess about the shape of the volume. Therefore, the precision of the volume estimates that one obtains from the method is only as good as the model and this cannot be ascertained from the single set of acquired data. In this letter I explain the inadequacies of the modelling method. I suggest that it be used only with caution, if at all. Instead I suggest two alternative ways to predict the CE, one that is based upon a rule-of-thumb approach to the object's shape, and another that is based upon spectral analysis of the measurement function and that is easy to implement with available computer software. PMID- 15817059 TI - Full-field strain mapping by optical correlation of micrographs acquired during deformation. AB - Optical correlation is an emerging strain-mapping technique that allows full field surface strain mapping by comparing the images of the same region before, during and after deformation. The fundamental aspects of optical correlation are presented, with emphasis on the applicability of the technique to the analysis of micrographs obtained during in situ deformation studies. Without considering specific algorithms, this paper discusses important practical issues such as accuracy and spatial resolution and how these are affected by image quality and other experimental difficulties. The technique was used to analyse image sequences obtained during in situ deformation tensile tests on two very different materials: antler bone and ferritic steel. As the technique does not require patterns or coatings to be applied on the surface of interest, the strain maps obtained could be used to relate strain heterogeneity to the underlying microstructure. PMID- 15817060 TI - Simultaneous degradation estimation and restoration of confocal images and performance evaluation by colocalization analysis. AB - A novel method for joint restoration and estimation of the degradation of confocal microscope images is presented. The observed images are degraded due to two sources: blurring due to the band-limited nature of the optical system [modelled by the point spread function (PSF)], and Poisson noise contaminates the observations due to the discrete nature of the photon detection process. The proposed method iterates noise reduction, blur estimation and deblurring, and applies these steps in two phases, i.e. a training phase and a restoration phase. In the first phase, these three steps are iterated until the blur estimation converges. Noise reduction and blur estimation are performed using steerable pyramids, and the deblurring is performed by the Richardson-Lucy algorithm. The second phase is the actual restoration. From then on, the blur estimation is used as a criterion to measure the image quality. The iterations are stopped when this measure converges, a result that is guaranteed. The integrated method is completely automatic, and no prior information on the image is required. The method has been given the name SPERRIL (Steerable Pyramid-based Estimation and Regularized Richardson-Lucy restoration). Compared with existing techniques by both objective measures and visual observation, in the SPERRIL-restored images noise is better suppressed. PMID- 15817061 TI - Imaging orientational order and lipid density in multilamellar vesicles with multiplex CARS microscopy. AB - Multiplex coherent anti-Stokes Raman scattering (CARS) microscopy is used to measure the width of the orientational order distribution of lipid acyl chains within a three-dimensionally confined microscopic probing volume. A theoretical model is developed to describe and simulate the polarization-dependent measurements. We observe that the orientational order in phosphatidylcholine multilammellar vesicles increases significantly upon addition of small amounts (95%). An oligonucleotide probe was designed to corroborate the assignment of the retrieved sequence to the symbiont and to detect similar bacteria rapidly. Antibiotic experiments showed that the elimination of the bacteria stops the reproductive cycle in E. harpa, as has been shown for the freshwater Euplotes species. PMID- 15817117 TI - Microgemma vivaresi n. sp. (Microsporidia, Tetramicridae), infecting liver and skeletal muscle of sea scorpions, Taurulus bubalis (Euphrasen 1786) (Osteichthyes, Cottidae), an inshore, littoral fish. AB - The ultrastructure of a new microsporidian species Microgemma vivaresi n. sp. causing liver cell xenoma formation in sea scorpions, Taurulus bubalis, is described. Stages of merogony, sporogony, and sporogenesis are mixed in the central cytoplasm of developing xenomas. All stages have unpaired nuclei. Uninucleate and multinucleate meronts lie within vacuoles formed from host endoplasmic reticulum and divide by binary or multiple fission. Sporonts, no longer in vacuoles, deposit plaques of surface coat on the plasma membrane that cause the surface to pucker. Division occurs at the puckered stage into sporoblast mother cells, on which plaques join up to complete the surface coat. A final binary fission gives rise to sporoblasts. A dense globule, thought to be involved in polar tube synthesis, is gradually dispersed during spore maturation. Spores are broadly ovoid, have a large posterior vacuole, and measure 3.6 microm x 2.1 microm (fresh). The polar tube has a short wide anterior section that constricts abruptly, then runs posteriad to coil about eight times around the posterior vacuole with granular contents. The polaroplast has up to 40 membranes arranged in pairs mostly attached to the wide region of the polar tube and directed posteriorly around a cytoplasm of a coarsely granular appearance. The species is placed alongside the type species Microgemma hepaticusRalphs and Matthews 1986 within the family Tetramicridae, which is transferred from the class Dihaplophasea to the class Haplophasea, as there is no evidence for the occurrence of a diplokaryotic phase. PMID- 15817118 TI - Electron tomographic and ultrastructural analysis of the Cryptosporidium parvum relict mitochondrion, its associated membranes, and organelles. AB - Sporozoites of the apicomplexan Cryptosporidium parvum possess a small, membranous organelle sandwiched between the nucleus and crystalloid body. Based upon immunolabelling data, this organelle was identified as a relict mitochondrion. Transmission electron microscopy and tomographic reconstruction reveal the complex arrangement of membranes in the vicinity of this organelle, as well as its internal organization. The mitochondrion is enveloped by multiple segments of rough endoplasmic reticulum that extend from the outer nuclear envelope. In tomographic reconstructions of the mitochondrion, there is either a single, highly-folded inner membrane or multiple internal subcompartments (which might merge outside the reconstructed volume). The infoldings of the inner membrane lack the tubular "crista junctions" found in typical metazoan, fungal, and protist mitochondria. The absence of this highly conserved structural feature is congruent with the loss, through reductive evolution, of the normal oxidative phosphorylation machinery in C. parvum. It is proposed that the retention of a relict mitochondrion in C. parvum is a strategy for compartmentalizing away from the cytosol toxic ferrous iron and sulfide, which are needed for iron sulfur cluster biosynthesis, an essential function of mitochondria in all eukaryotes. PMID- 15817119 TI - Morphological and molecular investigations of Tubulinosema ratisbonensis gen. nov., sp. nov. (Microsporidia: Tubulinosematidae fam. nov.), a parasite infecting a laboratory colony of Drosophila melanogaster (Diptera: Drosophilidae). AB - A new species of microsporidia from Drosophila melanogaster was investigated by light and electron microscopy and by ribosomal RNA (rRNA) sequencing. This microsporidium and the previously described Nosema kingi and Nosema acridophagus have been transferred to the new genus Tubulinosema gen. nov. with the following characters: nuclei are in diplokaryotic arrangement during the life cycle. All stages are in direct contact with the host cell cytoplasm, slightly anisofilar polar tube with the last coils being smaller in diameter arranged in one or two rows on both sides of the diplokaryon and small tubuli on the surface of late meronts. Spores are oval or slightly pyriform. Thick endospore wall, thinner over anchoring disc. This new genus and the genus Brachiola have been placed in a new family Tubulinosematidae fam. nov. Phylogenetic analysis of small subunit rRNA sequences by different methods placed Tubulinosema spp. in one clade with the genus Brachiola forming its sister clade, which is distant from the clade containing the true Nosema spp. including Nosema bombycis. PMID- 15817120 TI - Ultrastructural study of encystation and excystation in Acanthamoeba castellanii. AB - Encystation and excystation of Acanthamoeba castellanii were studied by transmission electron microscopy. The differentiation process was induced in asynchronous cultures grown axenically. Cytoplasmic vesicles containing a dense fibrous material very similar in appearance to the cyst wall were observed in trophozoites induced to encyst. When these trophozoites were incubated with calcofluor white m2r, fluorescence was observed in cytoplasmic vesicles, suggesting that the material contained in these vesicles corresponded to cyst wall precursors. Semithin cryosections of mature cysts with the same treatment showed fluorescence in the ectocyst and a less intense fluorescence in the endocyst, suggesting the presence of cellulose in both structures of the cyst wall. In mature cysts induced to excystation, small structures very similar to electron-dense granules (EDG) previously described in other amoebae were frequently observed. The EDGs were either sparsely distributed in the cytoplasm or associated with the cytoplasmic face of the plasma membrane. Many of them were located near the ostiole. In advanced phases of excystation, endocytic activity was suggested by the formation of endocytic structures and the presence of vacuoles with fibrous content similar to that of the cyst wall. Electron-dense granules in the process of dissolution were also observed in these vacuoles. Furthermore, the formation of a pseudopod suggests a displacement of the amoeba toward the ostiole. PMID- 15817114 TI - Protistan diversity estimates based on 18S rDNA from seawater incubations in the Western North Atlantic. AB - Cloning/sequencing and fragment analysis of ribosomal RNA genes (rDNA) are becoming increasingly common methods for the identification of microbial taxa. Sequences of these genes provide many additional taxonomic characters for species that otherwise have few distinctive morphological features, or that require involved microscopy or laboratory culture and testing. These same approaches are now being applied with great success in ecological studies of natural communities of microorganisms. Extensive information on the composition of natural microbial assemblages is being amassed at a rapid pace through genetic analyses of environmental samples and comparison of the resulting genetic information with well-established (and rapidly growing) public databases. We examined microbial eukaryote diversity in a natural seawater sample from the coastal western North Atlantic Ocean using two molecular biological approaches: the cloning and sequencing of rRNA genes and by fragment analysis of these genes using the terminal restriction fragment length polymorphism (T-RFLP) method. A simple experiment was carried out to examine changes in the overall eukaryote (largely protistan) diversity and species composition (phylotype diversity) of a natural microbial assemblage when a seawater sample is placed in a container and incubated at ambient light and temperature for 72 h. Containment of the natural seawater sample resulted in relatively minor changes in the overall eukaryote diversity (species richness) obtained by either molecular method at three time points (time-zero, time-24 h, time-72 h). However, substantial changes in the dominance of particular eukaryote phylotypes took place between the three sampling times. Only 18% of the total number of phylotypes observed in the study were observed at all three time points, while 65% (108 of 165) phylotypes were observed only at a single time point (54 unique phylotypes initially, 37 more unique phylotypes at 24 h, and 17 more at 72 h). The results of this study indicate that a high diversity of protistan taxa existed in the original seawater sample at very low abundance, and thus were not observed in the initial characterization of community structure. Containment resulted in significant shifts in the dominance of these taxa, enabling the presence of previously unobserved phylotypes to be documented after 24 or 72 h of incubation. PMID- 15817121 TI - Taxonomic characterization of two marine peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp. (Ciliophora: Peritrichia), from North China. AB - The morphology, infraciliature, and silverline system of two new peritrichous ciliates, Pseudovorticella clampi n. sp. and Zoothamnium pararbuscula n. sp., have been investigated based on both living and silver-impregnated specimens. Partial sequence of 18S-ITS1-5.8S rDNA of Z. pararbuscula is also determined in order to compare it with the closely related congener, Zoothamnium arbuscula. Zoothamnium pararbuscula can be distinguished from its close form Z. arbuscula by the different habitats, the appearance of the main stalk, the position of the contractile vacuole, and the information derived from 18S-ITS1-5.8S rDNA sequence analysis. Pseudovorticella clampi n. sp. is distinguished from its congeners by its body shape and size, pellicle granules, habitat, and number of transverse silverlines. PMID- 15817122 TI - Frontiers in genomics: insights into protist evolutionary biology, University of Iowa, May 19-21, 2004. AB - Protists constitute the bulk of eukaryotic diversity yet their genomes remain relatively unexplored. To address this issue, a workshop entitled, "Frontiers in Genomics: Insights into Protist Evolutionary Biology", was convened at the University of Iowa on June 19-21, 2004. The specific aims of the workshop were to define the role of genomics in the eukaryotic tree of life, to identify challenges in characterizing protist (i.e. microbial eukaryote) genomes, and in proposing specific solutions to these challenges. The findings of the workshop are presented here and in a white paper that provide a set of guidelines for organizing the protist community and for planning and executing a protist genome project. PMID- 15817124 TI - Combined Utilisation of Rapid Assessment Procedures for Loiasis (RAPLOA) and Onchocerciasis (REA) in Rain forest Villages of Cameroon. AB - BACKGROUND: Individuals with high microfilarial loads of Loa loa are at increased risk of neurologic serious adverse (SAE) events following ivermectin treatment against onchocerciasis. RAPLOA (Rapid Assessment Procedure for loiasis), a newly developed rapid assessment procedure for loiasis that relates the prevalence of key clinical manifestation of loiasis (history of eye worm) to the level of endemicity of the infection (prevalence of high intensity), is a very useful tool to identify areas at potential risk of L. loa post ivermectin treatment encephalopathy. In a perspective of treatment decision making in areas of co endemicity of loiasis/onchocerciasis, it would be advantageous (both in time and cost savings) for national onchocerciasis control programmes to use RAPLOA and the Rapid epidemiologic assessment for onchocerciasis (REA), in combination in given surveys. Since each of the two rapid assessment tools have their own specificities, the workability of combining the two methods needed to be tested. METHODS: We worked in 10 communities of a forest area presumed co-endemic for loiasis and onchocerciasis in the North-West Province of Cameroon where the mass treatment with ivermectin had not been carried out. A four-step approach was used and comprised: (i) generating data on the prevalence and intensity of loiasis and onchocerciasis in an area where such information is scarce; (ii) testing the relationship between the L. loa microfilaraemia prevalence and the RAPLOA prevalence, (iii) testing the relationship between the O. volvulus microfiladermia prevalence and the REA prevalence, (iv) testing the workability of combining RAPLOA/REA by study teams in which a single individual can perform the interview for RAPLOA and the nodule palpation for REA. RESULTS: The microfilaraemia prevalence of loiasis in communities ranged from 3.6% to 14.3%. 6 (0.61%) individuals had L. loa microfilarial loads above 8000 mf/ml but none of them attained 30,000 mf/ml, the threshold value above which the risk of developing neurologic SAE after ivermectin treatment is very high. None of the communities surveyed had RAPLOA prevalence above 40%. All the communities had microfiladermia prevalence above 60%. The microfiladermia results could be confirmed by the rapid epidemiologic method (nodule palpation), with all the 10 communities having REA prevalence above 20%. For the first time, this study has demonstrated that the two rapid assessment procedures for loiasis and onchocerciasis can be carried out simultaneously by a survey team, in which a single individual can administer the questionnaire for RAPLOA and perform the nodule palpation for REA. CONCLUSION: This study has: (i) Revealed that the Momo valley of the North West province of Cameroon is hyperendemic for onchocerciasis, but is of lower level of endemicity for L. loa. (ii) Confirmed the previous relationships established between RAPLOA and the L. loa microfilaraemia prevalence in one hand and between the REA and the O. volvulus microfiladermia prevalence in another hand (iii) Shown that RAPLOA and REA could be used simultaneously for the evaluation of loiasis and onchocerciasis endemicity in areas targeted by the African Programme for onchocerciasis Control for community directed treatment with ivermectin (CDTI). PMID- 15817123 TI - Enhanced levels of Hsulf-1 interfere with heparin-binding growth factor signaling in pancreatic cancer. AB - Hsulf-1 is a newly identified enzyme, which has the ability to decrease the growth of hepatocellular, ovarian, and head and neck squamous cell carcinoma cells by interfering with heparin-binding growth factor signaling. Since pancreatic cancers over-express a number of heparin-binding growth factors and their receptors, the expression and function of this enzyme in pancreatic cancer was analyzed. RESULTS: Pancreatic cancer samples expressed significantly (22.5 fold) increased Hsulf-1 mRNA levels compared to normal controls, and Hsulf-1 mRNA was localized in the cancer cells themselves as well as in peritumoral fibroblasts. 4 out of 8 examined pancreatic cancer cell lines expressed Hsulf-1, whereas its expression was below the level of detection in the other cell lines. Stable transfection of the Hsulf-1 negative Panc-1 pancreatic cancer cell line with a full length Hsulf-1 expression vector resulted in increased sulfatase activity and decreased cell-surface heparan-sulfate proteoglycan (HSPG) sulfation. Hsulf-1 expression reduced both anchorage-dependent and -independent cell growth and decreased FGF-2 mediated cell growth and invasion in this cell line. CONCLUSION: High expression of Hsulf-1 occurs in the stromal elements as well as in the tumor cells in pancreatic cancer and interferes with heparin binding growth factor signaling. PMID- 15817125 TI - A freely accessible, evidence based, objective system of analysis of posterior capsular opacification; evidence for its validity and reliability. AB - BACKGROUND: The aim of this study was to develop a system of computerised analysis of digital images of posterior capsule opacification (PCO) that is evidence based, objective and freely available. The paper will present evidence for the reliability and validity of the developed system. METHODS: The system of PCO analysis was developed considering current published evidence on visual significance of PCO and additional investigative analysis of PCO images. Details of the image processing and analysis steps are discussed and a final system that measures an entropy score weighted toward proximity to central areas is described. In order to assess validity, the systems ability to measure PCO progression is assessed along with the visual significance of its final computerised scores. Reliability of the system is also assessed. RESULTS: The final system runs successfully and is simple to use. Analyses of PCO by the system show an ability to detect early progression of PCO as well as detection of visually significant PCO. Images with no clinical PCO produce very low scores in the analysis. Reliability of the system of analysis is shown to be satisfactory. CONCLUSION: This paper presents a system of PCO analysis that is evidence based, objective and clinically useful. Substantial evidence is provided for its validity and reliability. PMID- 15817126 TI - Valine 738 and lysine 735 in the fifth transmembrane domain of rTas1r3 mediate insensitivity towards lactisole of the rat sweet taste receptor. AB - BACKGROUND: The sweet taste inhibitor lactisole acts on the human sweet taste receptor heteromer TAS1R2-TAS1R3 but not on its rodent counterpart. Recently, it was shown that the lactisole sensitivity of the human sweet taste receptor involves the part of TAS1R3 encompassing the seven transmembrane regions but not the huge N-terminal domain. Using mutational analysis we investigated which amino acid residues distinguish lactisole insensitive rat from sensitive human T1R3 receptors. RESULTS: The functional analysis of specific receptor mutants in HEK293T cells revealed that the exchange of valine 738 in the fifth transmembrane domain of rTas1r3 by an alanine is sufficient to confer lactisole sensitivity to the rat sweet taste receptor. The sensitivity of this receptor mutant is approximately 2 fold lower than the sensitivity of the human sweet taste receptor. Additional substitution of lysine 735 by phenylalanine in rTas1r3 results in a rat sweet taste receptor that is as sensitive to lactisole as its human counterpart. The exchange of valine 738 to alanine was accompanied by a approximately 50% reduction in receptor efficacy. This effect was seen with all six different sweet compounds examined. CONCLUSION: The lactisole insensitivity of rat sweet taste receptor is caused by only two amino acids in transmembrane region five, which is critical for the interaction of lactisole with the sweet taste receptor. The observation that the mutant receptor simultaneously displays a generally reduced sensitivity towards all agonists suggests that the lactisole insensitivity of the rodent receptor might be more likely caused by the inaccessibility of the lactisole binding site rather then by its direct disruption. PMID- 15817127 TI - Combination of qualitative and quantitative methods for developing a new Health Related Quality of Life measure for patients with anogenital warts. AB - BACKGROUND: Anogenital warts are the most easily recognized sign of genital Human Papilloma Virus infection. The objective was to develop a short, valid and reliable questionnaire to measure Health Related Quality of Life (HRQL) in patients with anogenital warts. METHODS: First a literature review was performed to identify relevant papers describing the impact of anogenital warts in HRQL; second the main domains were identified by some experts in a focus group, and third in-depth-semi-structured interviews were conducted in patients with anogenital warts to identify the initial set of items. A qualitative reduction of the initial set of items was performed based on the mean scoring of the experts for the three scales: clarity, frequency and importance. The initial questionnaire was pilot tested in 135 patients. Rasch analysis was performed with the results of the questionnaire in order to refine the instrument. Spearman's correlation was calculated between the initial questionnaire and the reduced version. Additionally the measurement properties (validity and reliability) of the resulting final questionnaire were tested and compared using standard procedures (Cronbach's Alpha and item-total correlation). RESULTS: the main domains identified as affected in patient's life were: sexual, colleagues and partner relationships. After a proper qualitative reduction the initial set of 134 items was reduced to 22. The questionnaire was pilot tested in 135 patients and two dimensions were identified after the multifactorial analysis: emotional dimension and sexual activity dimension. As a result of the Rasch analysis the questionnaire was reduced to 10 items. High correlation was found between the initial and the reduced version for the two dimensions. Cronbach's alpha values were acceptable (0.86). CONCLUSION: The initial 22 items questionnaire was reduced by Rasch analysis to a version of 10 items, with two dimensions: emotional and sexual. The results suggest the adequacy of the 10 items to evaluate HRQL of patients with anogenital warts in a valid and reliable way. PMID- 15817128 TI - A method for aligning RNA secondary structures and its application to RNA motif detection. AB - BACKGROUND: Alignment of RNA secondary structures is important in studying functional RNA motifs. In recent years, much progress has been made in RNA motif finding and structure alignment. However, existing tools either require a large number of prealigned structures or suffer from high time complexities. This makes it difficult for the tools to process RNAs whose prealigned structures are unavailable or process very large RNA structure databases. RESULTS: We present here an efficient tool called RSmatch for aligning RNA secondary structures and for motif detection. Motivated by widely used algorithms for RNA folding, we decompose an RNA secondary structure into a set of atomic structure components that are further organized by a tree model to capture the structural particularities. RSmatch can find the optimal global or local alignment between two RNA secondary structures using two scoring matrices, one for single-stranded regions and the other for double-stranded regions. The time complexity of RSmatch is O(mn) where m is the size of the query structure and n that of the subject structure. When applied to searching a structure database, RSmatch can find similar RNA substructures, and is capable of conducting multiple structure alignment and iterative database search. Therefore it can be used to identify functional RNA motifs. The accuracy of RSmatch is tested by experiments using a number of known RNA structures, including simple stem-loops and complex structures containing junctions. CONCLUSION: With respect to computing efficiency and accuracy, RSmatch compares favorably with other tools for RNA structure alignment and motif detection. This tool shall be useful to researchers interested in comparing RNA structures obtained from wet lab experiments or RNA folding programs, particularly when the size of the structure dataset is large. PMID- 15817129 TI - Building a protein name dictionary from full text: a machine learning term extraction approach. AB - BACKGROUND: The majority of information in the biological literature resides in full text articles, instead of abstracts. Yet, abstracts remain the focus of many publicly available literature data mining tools. Most literature mining tools rely on pre-existing lexicons of biological names, often extracted from curated gene or protein databases. This is a limitation, because such databases have low coverage of the many name variants which are used to refer to biological entities in the literature. RESULTS: We present an approach to recognize named entities in full text. The approach collects high frequency terms in an article, and uses support vector machines (SVM) to identify biological entity names. It is also computationally efficient and robust to noise commonly found in full text material. We use the method to create a protein name dictionary from a set of 80,528 full text articles. Only 8.3% of the names in this dictionary match SwissProt description lines. We assess the quality of the dictionary by studying its protein name recognition performance in full text. CONCLUSION: This dictionary term lookup method compares favourably to other published methods, supporting the significance of our direct extraction approach. The method is strong in recognizing name variants not found in SwissProt. PMID- 15817130 TI - Add-on topiramate reduces weight in overweight patients with affective disorders: a clinical case series. AB - BACKGROUND: The weight-gain caused by many psychotropic drugs is a major cause for poor compliance with such medications and could also increase cardio-vascular morbidity among psychiatric patients. Recent reports have shown that the anticonvulsant topiramate causes weight loss in various patient groups. The drug has also shown effectiveness in open trials as a mood stabilizer in patients with affective disorders, but not in controlled trials in the acute treatment of mania. We used topiramate to treat 12 patients with affective disorders who had a body-mass index > 30 kg/m2. METHODS: Topiramate was prescribed as part of our routine clinical practice, as an add-on medication, or as a replacement of a mood stabilizer. Patients' weight was recorded in 1 to 2 monthly intervals. Patients were followed up for between 6 and 12 months. The final dose of topiramate varied from 200 to 600 mg/day. RESULTS: Topiramate was effective in reducing the weight in 10 out of the 12 patients. At six months the 12 patients had lost a mean of 7.75 kg (SD = 6.9 kg, p < 0.001) and at 12 months 9 patients had lost a mean of 9.61 kg (SD = 6.7 kg, p = 0.003). Three patients stopped the treatment: one due to side effects, one due to possible side effects, and one suffered a manic relapse and showed no sustained weight loss. There were no other clear changes in the course of illness of the patients. CONCLUSION: The evidence of a strong weight-reducing potential of topiramate is indisputable and clinically significant. Topiramate could be considered in the treatment of bipolar patients who are overweight, or whose concerns about weight gain compromise their compliance with long-term prophylactic medication. So far there is no evidence that topiramate has anti-manic effect and it should not be used as monotherapy. PMID- 15817131 TI - Exogenous recombinant human growth hormone effects during suboptimal energy and zinc intake. AB - BACKGROUND: Energy and Zinc (Zn) deficiencies have been associated with nutritional related growth retardation as well as growth hormone (GH) resistance. In this study, the relationship between suboptimal energy and/or Zn intake and growth in rats and their response to immunoreactive exogenous recombinant human GH (GHi), was determined. RESULTS: Rats treated with GHi and fed ad-libitum energy and Zn (100/100) had increased IGFBP-3 (p < 0.05) as compared with NSS (215 +/- 23 vs. 185 +/- 17 ng/ml) along with similar body weight gain. Rats treated with GHi and fed suboptimal energy and full Zn (70/100) had significantly increased weight gain (109.0 +/- 18.2 vs. 73.8 +/- 11.0 g) and serum IGF-I levels (568 +/- 90 vs. 420 +/- 85 ng/ml), along with decreased total body water (TBW; 61.0 +/- 1.6 vs. 65.7 +/- 2.1%) as compared to NSS controls. However, body weight gain was reduced (p < 0.05) as compared with rats fed ad-libitum energy. Growth hormone treated rats fed only suboptimal Zn (100/70), had increased weight gain (217.5 +/- 13.2 vs. 191.6 +/- 17.9 g; p < 0.05) compared to those given NSS. These rats gained weight in similar amounts to those fed full Zn. Rats treated with GHi and fed both suboptimal energy and Zn (70/70) showed similar results to those fed suboptimal energy with appropriate Zn (70/100), along with significant increases in IGFBP-3 levels (322 +/- 28 vs. 93 +/- 28 ng/ml). All restricted rats had reduced 24-h EE (kcal/100 g BW) and physical activity index (oscillations/min/kg BW) and GHi did not overcome these effects. CONCLUSION: These results suggest that GHi enhances weight gain in rats with suboptimal energy and Zn intake but does not modify energy expenditure or physical activity index. Suboptimal Zn intake did not exacerbate the reduced growth or decrease in energy expenditure observed with energy restriction. PMID- 15817132 TI - Diseased muscles that lack dystrophin or laminin-alpha2 have altered compositions and proliferation of mononuclear cell populations. AB - BACKGROUND: Multiple types of mononucleate cells reside among the multinucleate myofibers in skeletal muscles and these mononucleate cells function in muscle maintenance and repair. How neuromuscular disease might affect different types of muscle mononucleate cells had not been determined. In this study, therefore, we examined how two neuromuscular diseases, dystrophin-deficiency and laminin-alpha2 deficiency, altered the proliferation and composition of different subsets of muscle-derived mononucleate cells. METHODS: We used fluorescence-activated cell sorting combined with bromodeoxyuridine labeling to examine proliferation rates and compositions of mononuclear cells in diseased and healthy mouse skeletal muscle. We prepared mononucleate cells from muscles of mdx (dystrophin-deficient) or Lama2-/- (laminin-alpha2-deficient) mice and compared them to cells from healthy control muscles. We enumerated subsets of resident muscle cells based on Sca-1 and CD45 expression patterns and determined the proliferation of each cell subset in vivo by BrdU incorporation. RESULTS: We found that the proliferation and composition of the mononucleate cells in dystrophin-deficient and laminin alpha2-deficient diseased muscles are different than in healthy muscle. The mdx and Lama2-/- muscles showed similar significant increases in CD45+ cells compared to healthy muscle. Changes in proliferation, however, differed between the two diseases with proliferation increased in mdx and decreased in Lama2-/- muscles compared to healthy muscles. In particular, the most abundant Sca-1-/CD45- subset, which contains muscle precursor cells, had increased proliferation in mdx muscle but decreased proliferation in Lama2-/- muscles. CONCLUSION: The similar increases in CD45+ cells, but opposite changes in proliferation of muscle precursor cells, may underlie aspects of the distinct pathologies in the two diseases. PMID- 15817133 TI - Differential effect of obesity on bone mineral density in White, Hispanic and African American women: a cross sectional study. AB - Osteoporosis is a major public health problem with low bone mass affecting nearly half the women aged 50 years or older. Evidence from various studies has shown that higher body mass index (BMI) is a protective factor for bone mineral density (BMD). Most of the evidence, however, is from studies with Caucasian women and it is unclear to what extent ethnicity plays a role in modifying the effect of BMI on BMD.A cross sectional study was performed in which records of postmenopausal women who presented for screening for osteoporosis at 2 urban medical centres were reviewed. Using logistic regression, we examined the interaction of race and BMI after adjusting for age, family history of osteoporosis, maternal fracture, smoking, and sedentary lifestyle on BMD. Low BMD was defined as T-score at the lumbar spine < -1.Among 3,206 patients identified, the mean age of the study population was 58.3 +/- 0.24 (Years +/- SEM) and the BMI was 30.6 kg/m2. 2,417 (75.4%) were African Americans (AA), 441(13.6%) were Whites and 348 (10.9%) were Hispanics. The AA women had lower odds of having low BMD compared to Whites [Odds ratio (OR) = 0.079 (0.03-0.24) (95% CI), p < 0.01]. The odds ratio of low BMD was not statistically significant between White and Hispanic women. We examined the interaction between race and BMD. For White women; as the BMI increases by unity, the odds of low BMD decreases [OR = 0.9 (0.87-0.94), p < 0.01; for every unit increase in BMI]. AA women had slightly but significantly higher odds of low BMD compared to Whites [OR 1.015 (1.007-1.14), p <0.01 for every unit increase in BMI]. This effect was not observed when Hispanic women were compared to Whites.There is thus a race-dependent effect of BMI on BMD. With each unit increase in BMI, BMD increases for White women, while a slight but significant decrease in BMD occurs in African American women. PMID- 15817134 TI - Individual sequences in large sets of gene sequences may be distinguished efficiently by combinations of shared sub-sequences. AB - BACKGROUND: Most current DNA diagnostic tests for identifying organisms use specific oligonucleotide probes that are complementary in sequence to, and hence only hybridize with the DNA of one target species. By contrast, in traditional taxonomy, specimens are usually identified by 'dichotomous keys' that use combinations of characters shared by different members of the target set. Using one specific character for each target is the least efficient strategy for identification. Using combinations of shared bisectionally-distributed characters is much more efficient, and this strategy is most efficient when they separate the targets in a progressively binary way. RESULTS: We have developed a practical method for finding minimal sets of sub-sequences that identify individual sequences, and could be targeted by combinations of probes, so that the efficient strategy of traditional taxonomic identification could be used in DNA diagnosis. The sizes of minimal sub-sequence sets depended mostly on sequence diversity and sub-sequence length and interactions between these parameters. We found that 201 distinct cytochrome oxidase subunit-1 (CO1) genes from moths (Lepidoptera) were distinguished using only 15 sub-sequences 20 nucleotides long, whereas only 8-10 sub-sequences 6-10 nucleotides long were required to distinguish the CO1 genes of 92 species from the 9 largest orders of insects. CONCLUSION: The presence/absence of sub-sequences in a set of gene sequences can be used like the questions in a traditional dichotomous taxonomic key; hybridisation probes complementary to such sub-sequences should provide a very efficient means for identifying individual species, subtypes or genotypes. Sequence diversity and sub-sequence length are the major factors that determine the numbers of distinguishing sub-sequences in any set of sequences. PMID- 15817135 TI - Atomoxetine blocks motor hyperactivity in neonatal 6-hydroxydopamine-lesioned rats: implications for treatment of attention-deficit hyperactivity disorder. AB - We recently reported that selective inhibitors of neuronal transport of norepinephrine (NE), desipramine and nisoxetine, reversed motor hyperactivity in an animal model of attention-deficit hyperactivity disorder (ADHD). In this study, we examined behavioural effects of atomoxetine, a potent new NE reuptake blocker, in juvenile male rats with neonatal 6-hydroxydopamine (6-OHDA) lesions of dopamine projections to the forebrain. 6-OHDA (100 microg) was administered intracisternally on postnatal day (PD) 5 following desipramine (25 mg/kg s.c.) pretreatment to protect noradrenergic neurons. Atomoxetine (1 mg/kg) was given intraperitoneally before recording motor activity for 90 min at PD 23-26 in a novel environment. Atomoxetine greatly reduced motor hyperactivity in 6-OHDA lesioned rats while exhibiting transient sedative effects in sham controls. The observed effects in this animal model for ADHD are consistent with the emerging clinical use of atomoxetine as a novel, non-stimulant treatment for ADHD. PMID- 15817136 TI - Prediction of the ability of clozapine to treat negative symptoms from plasma glycine and serine levels in schizophrenia. AB - We previously reported that plasma levels of glycine, a co-agonist at N-methyl-D asparate (NMDA)-type glutamate receptors, are decreased in patients with schizophrenia, and that glycine levels are negatively correlated with negative symptoms. The aim of the present study was to determine if glycine, or its ratio to serine, a precursor of glycine, predicts change in negative symptoms in subjects with schizophrenia during treatment with clozapine, an atypical antipsychotic drug with multiple effects on glutamatergic activity. Plasma levels of glycine, serine, and their ratio, were measured in 44 patients with schizophrenia who were subsequently treated with clozapine. Baseline glycine levels or glycine/serine ratios predicted the Scale for the Assessment of Negative Symptoms - Sum of the Global Scales and Avolition-Apathy after 6 wk of clozapine treatment. These results indicate the association of these amino acid measures with response to clozapine in terms of negative symptoms in patients with schizophrenia. PMID- 15817137 TI - Bupropion SR for the treatment of postpartum depression: a pilot study. AB - Despite the prevalence of postpartum depression, few studies have assessed the efficacy of antidepressants for the treatment of this disorder. Failure to treat postpartum depression (PPD) places the woman at risk for chronic depression and may have adverse effects on child wellbeing and development. Eight female outpatients aged 18-45 yr were enrolled in an 8-wk open-label trial of bupropion SR for PPD. All patients met DSM-IV criteria for major depression with onset within 3 months of delivery and scored 17 or greater on the Hamilton Depression Rating Scale (HAMD) at baseline. Those with onset of depressive symptoms during pregnancy, psychotic symptoms, or significant medical illness were excluded. Median scores on the HAMD declined from 20.5 (range 15-38) at baseline to 10.0 (range 1-20) at end-point (p<0.05, Wilcoxon signed-ranks test; LOCF). Six out of the eight subjects demonstrated a > or =50% decrease in HAMD scores from baseline; three subjects achieved remission (HAMD score of < or =7) at week 8. Median final dosage of bupropion SR was 262.5 (range 37.5-300). Bupropion SR was well tolerated, and no subjects discontinued treatment as a result of medication side-effects. Bupropion SR represents an effective and well-tolerated antidepressant for the treatment of PPD. PMID- 15817138 TI - Limitations to the measurement of oxygen concentrations by HRTEM imposed by surface roughness. PMID- 15817140 TI - Electron microscopy study of exotic nanostructures of cadmium sulfide. AB - In this article, two simple methods, evaporation-condensation and catalytic thermal evaporation, were used to investigate the synthesis of CdS nanostructures for nanoscale optoelectronic applications. To understand their growth mechanisms, various electron microscopy and microanalysis techniques were utilized in characterizing their morphologies, internal structures, growth directions and elemental compositions. The electron microscopy study reveals that when using the evaporation-condensation method, branched CdS nanorods and self-assembled arrays of CdS nanorods were synthesized at 800 degrees C and 1000 degrees C, respectively. Instead of morphological differences, both types of CdS nanorods grew along the [0001] direction. However, when using the catalytic thermal evaporation method (Au as the catalyst), patterned CdS nanowires and nanobelts were formed at the temperature region of 500-600 degrees C and 600-750 degrees C, respectively. Their growth direction was along the direction [1010] instead of [0001]. Based on the microscopy and microanalysis results, we propose some growth mechanisms in relation to the growth processes of those exotic CdS nanostructures. PMID- 15817141 TI - A scanning electron and atomic force microscopy study of the surface morphology and composition of CsI films as affected by evaporation rate and humid-air exposure. AB - Evaporation rate and subsequent exposure to humid air affect the surface morphology and composition of cesium iodide (CsI) films and, in turn, their photoemissive efficiency when used as photocathodes. The surface morphology and elemental composition of 300-nm-thick CsI films grown at two different rates (1 nm/s and 0.04 nm/s), both freshly evaporated and after 24-h exposure to humid air were investigated by means of atomic force microscopy and scanning electron microscopy/electron diffraction spectroscopy. The CsI film freshly evaporated at a slow rate exhibited a granular surface presenting circular holes or craters where the CsI material was moved from the center to the boundaries. After 24-h exposure to humid air, this film coalesced in large grain showing a marked increase of surface roughness. Conversely, the CsI film grown at a fast rate mostly retained its original surface uniformity and homogeneity with no presence of holes and craters after 24-h exposure to humid air. Further, surface roughness and average peak height decreased, but the surface coalesced in large grains spaced by small fractures where the CsI coverage was almost lost. In conclusion, the films grown at a fast evaporation rate were affected by 24-h exposure to humid air less than those grown at a slow rate, and are thus expected to possess a greater long-term stability. PMID- 15817142 TI - Transmission electron microscopy and X-Ray diffraction analysis of aluminum induced crystallization of amorphous silicon in alpha-Si:H/Al and Al/alpha-Si:H structures. AB - Solid phase crystallization of plasma-enhanced chemical-vapor-deposited (PECVD) amorphous silicon (alpha-Si:H) in alpha-Si:H/Al and Al/alpha-Si:H structures has been investigated using transmission electron microscopy (TEM) and X-ray diffraction (XRD). Radiative heating has been used to anneal films deposited on carbon-coated nickel (Ni) grids at temperatures between 200 and 400 degrees C for TEM studies. alpha-Si:H films were deposited on c-Si substrates using high vacuum (HV) PECVD for the XRD studies. TEM studies show that crystallization of alpha Si:H occurs at 200 degrees C when Al film is deposited on top of the alpha-Si:H film. Similar behavior was observed in the XRD studies. In the case of alpha-Si:H deposited on top of Al films, the crystallization could not be observed at 400 degrees C by TEM and even up to 500 degrees C as seen by XRD. PMID- 15817143 TI - The distribution of light elements in biological cells measured by electron probe X-ray microanalysis of cryosections. AB - The intracellular distribution of the elements carbon, nitrogen, and oxygen was measured in cultured rat hepatocytes by energy dispersive electron probe X-ray microanalysis of 100-nm-thick freeze-dried cryosections. Electron irradiation with a dose up to 106 e/nm2 caused no or merely negligible mass loss in mitochondria and in cytoplasm. Cell nuclei lost carbon, nitrogen, and-to a clearly higher extent-oxygen with increasing electron irradiation. Therefore, electron doses less than 3 x 105 e/nm2 were used to measure the subcellular compartmentation of carbon, nitrogen, and oxygen in cytoplasm, mitochondria, and nuclei of the cells. The subcellular distribution of carbon, nitrogen, and oxygen reflects the intracellular compartmentation of various biomolecules. Cells exposed to inorganic mercury before cryofixation showed an increase of oxygen in nuclei and cytoplasm. Concomitantly the phosphorus/nitrogen ratio decreased in mitochondria. The data suggest mercury-induced production of ribonucleic acid (RNA) and decrease of adenosine triphosphate (ATP). Although biomolecules cannot be identified by X-ray microanalysis, measurements of the whole element spectrum including the light elements carbon, nitrogen, and oxygen can be useful to study specific biomolecular activity in cellular compartments depending on the functional state of the cell. PMID- 15817144 TI - The role of microfilaments in early meiotic maturation of mouse oocytes. AB - Mouse oocyte microfilaments (MF) were perturbed by depolymerization (cytochalasin B) or stabilization (jasplakinolide) and correlated meiotic defects examined by confocal microscopy. MF, microtubules, and mitochondria were vitally stained; centrosomes (gamma-tubulin), after fixation. MF depolymerization by cytochalasin in culture medium did not affect central migration of centrosomes, mitochondria, or nuclear breakdown (GVBD); some MF signal was localized around the germinal vesicle (GV). In maturation-blocking medium (containing IBMX), central movement was curtailed and cortical MF aggregations made the plasma membrane wavy. Occasional long MF suggested that not all MF were depolymerized. MF stabilization by jasplakinolide led to MF aggregations throughout the cytoplasm. GVBD occurred (unless IBMX was present) but no spindle formed. Over time, most oocytes constricted creating a dumbbell shape with MF concentrated under one-half of the oocyte cortex and on either side of the constriction. In IBMX medium, the MF containing half of the dumbbell over time sequestered the GV, MF, mitochondria, and one to two large cortical centrosomes; the non-MF half appeared empty. Cumulus processes contacted the oocyte surface (detected by microtubule content) and mirrored MF distribution. Results demonstrated that MF play an essential role in meiosis, primarily through cortically mediated events, including centrosome localization, spindle (or GV) movement to the periphery, activation of (polar body) constriction, and establishment of oocyte polarity. The presence of a cortical "organizing pole" is hypothesized. PMID- 15817145 TI - Microscopic examination of chitosan-polyphosphate beads with entrapped spores of the biocontrol agent, Streptomyces melanosporofaciens EF-76. AB - Spores of the biocontrol agent, Streptomyces melanosporofaciens EF-76, were entrapped by complex coacervation in beads composed of a macromolecular complex (MC) of chitosan and polyphosphate. A proportion of spores entrapped in beads survived the entrapment procedure as shown by treating spores from chitosan beads with a dye allowing the differentiation of live and dead cells. The spore-loaded chitosan beads could be digested by a chitosanase, suggesting that, once introduced in soil, the beads would be degraded to release the biocontrol agent. Spore-loaded beads were examined by optical and scanning electron microscopy because the release of the biological agent depends on the spore distribution in the chitosan beads. The microscopic examination revealed that the beads had a porous surface and contained a network of inner microfibrils. Spores were entrapped in both the chitosan microfibrils and the bead lacuna. PMID- 15817146 TI - Interrelations between the parasitophorous vacuole of Toxoplasma gondii and host cell organelles. AB - Toxoplasma gondii, the causative agent of toxoplasmosis, is capable of actively penetrating and multiplying in any nucleated cell of warm-blooded animals. Its survival strategies include escape from fusion of the parasitophorous vacuole with host cell lysosomes and rearrangement of host cell organelles in relation to the parasitophorous vacuole. In this article we report the rearrangement of host cell organelles and elements of the cytoskeleton of LLCMK2 cells, a lineage derived from green monkey kidney epithelial cells, in response to infection by T. gondii tachyzoites. Transmission electron microscopy made on flat embedded monolayers cut horizontally to the apical side of the cells or field emission scanning electron microscopy of monolayers scraped with scotch tape before sputtering showed that association of mitochondria to the vacuole is much less frequent than previously described. On the other hand, all parasitophorous vacuoles were surrounded by elements of the endoplasmic reticulum. These data were complemented by observations by laser scanning microscopy using fluorescent probes from mitochondria and endoplasmic reticulum and reinforced by three dimensional reconstruction from serial sections observed by transmission electron microscopy and labeling of mitochondria and endoplasmic reticulum by fluorescent probes. PMID- 15817147 TI - Immunolocalization of MAP-2 in routinely formalin-fixed, paraffin-embedded guinea pig brain sections using microwave irradiation: a comparison of different combinations of antibody clones and antigen retrieval buffer solutions. AB - The present study was designed to evaluate the efficacy of different microwave pretreatment methods to retrieve microtubule-associated protein 2 (MAP-2) immunoreactivity in formalin-fixed, paraffin-embedded guinea pig brain sections. Brain sections, microwave pretreated in boiling sodium citrate, citric acid, Tris hydrochloride, and EDTA buffers of pH 4, 6, and 8, were labeled with four different clones of MAP-2 monoclonal antibodies. No MAP-2 immunoreactivity was observed in control sections processed without microwave pretreatment. Optimal MAP-2 immunoreactivity was observed only when MAP-2 antibody clone AP18 was used in conjunction with citric acid buffer of pH 6.0. Using this combination, brain sections from nerve agent soman-exposed guinea pigs were found to exhibit marked reduction in MAP-2 immunostaining in the hippocampus. These observations suggest that the clone of the antibody in addition to the type and pH of antigen retrieval (AR) solution are important variables to be considered for establishing an optimal AR technique. When studying counterpart antigens of species other than that to which the antibodies were originally raised, different antibody clones must be tested in combination with different microwave-assisted AR (MAR) methods. This MAR method makes it possible to conduct retrospective studies on archival guinea pig brain paraffin blocks to evaluate changes in neuronal MAP-2 expression as a consequence of chemical warfare nerve agent toxicity. PMID- 15817150 TI - Decreased cell survival and DNA repair capacity after UVC irradiation in association with down-regulation of GRP78/BiP in human RSa cells. AB - In contrast to extensive studies on the roles of molecular chaperones, such as heat shock proteins, there are only a few reports about the roles of GRP78/BiP, an endoplasmic reticulum (ER) stress-induced molecular chaperone, in mammalian cell responses to DNA-damaging stresses. To investigate whether GRP78/BiP is involved in resistance to a DNA-damaging agent, UVC (principally 254 nm in wavelength), we established human cells with down-regulation of GRP78/BiP by transfection of human RSa cells with antisense cDNA for GRP78/BiP. We found that the transfected cells showed higher sensitivity to UVC-induced cell death than control cells transfected with the vector alone. In the antisense-cDNA transfected cells, the removal capacities of the two major types of UVC-damaged DNA (thymine dimers and (6-4) photoproducts) in vivo and DNA synthesis activity of whole cell extracts to repair UVC-irradiated plasmids in vitro were remarkably decreased compared with those in the control cells. Furthermore, the antisense cDNA transfected cells also showed slightly higher sensitivity to cisplatin induced cell death than the control cells. Cisplatin-induced DNA damage is primarily repaired by nucleotide excision repair, like UVC-induced DNA damage. The present results suggest that GRP78/BiP plays a protective role against UVC induced cell death possibly via nucleotide excision repair, at least in the human RSa cells tested. PMID- 15817149 TI - Expression of progesterone receptor B is associated with G0/G1 arrest of the cell cycle and growth inhibition in NIH3T3 cells. AB - Previously, we found a significant reduction of progesterone receptor B (PR-B) expression levels in the Ras-mediated NIH3T3 cell transformation, and re expression of exogenous PR-B eliminated the tumorigenic potential. We hypothesized that this reduction is of biological significance in cell transformation. In the present study, we determined the correlation between PR-B expression and cell cycle progression. In synchronized NIH3T3 cells, we found an increase in PR-B protein and p27 CDK inhibitor levels in the G0/G1 phase and a reduction due to redistribution in the S and G2/M phases. The MEK inhibitor or cAMP stimulation arrested NIH3T3 cells in the G0/G1 phase of the cell cycle. The expression of PR-B and p27 CDK inhibitors was up-regulated by treatment with both the MEK inhibitor and cAMP. Treatment of synchronized cells with a PKA inhibitor in the presence of 1% calf serum resulted in a significant reduction in both PR-B and p27 levels. The decrease in the PR-B levels caused by anti-sense oligomers or siRNA corresponded to the reduction in p27 levels. PR-B overexpression by adenovirus infection induced p27 and suppressed cell growth. Finally, we showed that PR-B modulation involved in the regulation of NIH3T3 cell proliferation was independent of nuclear estrogen receptor (ER) activity but dependent on non genomic ER activity. PMID- 15817151 TI - The catenin p120ctn inhibits Kaiso-mediated transcriptional repression of the beta-catenin/TCF target gene matrilysin. AB - The POZ-zinc finger transcription factor Kaiso was first identified as a specific binding partner for the Armadillo catenin and cell adhesion cofactor, p120ctn. Kaiso is a unique POZ protein with bi-modal DNA-binding properties; it associates with a sequence-specific DNA consensus Kaiso binding site (KBS) or methylated CpG dinucleotides, and regulates transcription of artificial promoters containing either site. Interestingly, the promoter of the Wnt/beta-catenin/TCF target gene matrilysin possesses two conserved copies of the KBS, which suggested that Kaiso might regulate matrilysin expression. In this study, we demonstrate using chromatin immunoprecipitation analysis that Kaiso associates with the matrilysin promoter in vivo. Minimal promoter assays further confirmed that Kaiso specifically repressed transcription of the matrilysin promoter; mutation of the KBS element or RNAi-mediated depletion of Kaiso abrogated this effect. More importantly, Kaiso blocked beta-catenin-mediated activation of the matrilysin promoter. Consistent with our previous findings, both Kaiso-DNA binding and Kaiso mediated transcriptional repression of the matrilysin promoter were inhibited by overexpression of wild-type p120ctn, but not by a p120ctn mutant exhibiting impaired nuclear import. Collectively, our data establish Kaiso as a sequence specific transcriptional repressor of the matrilysin promoter, and suggest that p120ctn and beta-catenin act in a synergistic manner, via distinct mechanisms, to activate matrilysin expression. PMID- 15817152 TI - The Ku70-binding site of Ku80 is required for the stabilization of Ku70 in the cytoplasm, for the nuclear translocation of Ku80, and for Ku80-dependent DNA repair. AB - Ku plays a key role in multiple nuclear processes, e.g., DNA repair, transcription regulation, and replication. It is believed that heterodimerization between Ku70 and Ku80 is essential for Ku-dependent DNA repair, although its role is poorly understood. We previously identified the Ku70-binding site of Ku80. In this study, to understand the role of heterodimerization in the function of Ku, we generated and/or analyzed cell lines stably expressing the EGFP-tagged-wild type human Ku80, its Ku70-binding mutant, its NLS-dysfunctional mutant, or its double mutant in Ku80-deficient cells. Our results show that the Ku70-binding site of Ku80 is required for the stabilization of Ku70 in the cytoplasm and for the nuclear translocation of Ku80 through its heterodimerization with Ku70. In addition, our results suggest that the nuclear translocation of Ku80 through the Ku70-binding site as well as through the NLS of Ku80 play, at least in part, a role in Ku80-dependent DNA repair. Furthermore, our results suggest the possibility that Ku80 has a DNA DSB repair function independent of Ku70 in the nuclei, in addition to that dependent on Ku70. PMID- 15817153 TI - Vascular smooth muscle cell polyploidization involves changes in chromosome passenger proteins and an endomitotic cell cycle. AB - Vascular smooth muscle cell polyploidization occurs during normal development and is enhanced under physiologic stress, but the mechanism of this cell cycle has not been explored. We show via time-lapse video imaging and immunofluorescence analyses that primary vascular smooth muscle cells (VSMC) undergo an endomitotic type cell cycle, including a normal progression through part of mitosis. Mononuclear polyploid cells are generated by defects in sister chromatid separation and/or segregation, and cellular binucleation occurs by reversal of cytokinesis. To obtain further leads to regulators involved, we examined the chromosomal passenger proteins, Aurora B, inner centromere protein and Survivin, and concluded that Aurora B and inner centromere protein are normally colocalized in centromeres, the midzone, and the midbody during mitosis. Survivin, however, is dim and diffused; it does not colocalize with either Aurora B or inner centromere protein in VSMC, which could account for defects in sister chromatid separation and/or segregation and reversal of cytokinesis. In accordance with the reported dependency of Aurora B activity on Survivin, the Aurora B substrate, vimentin, is not phosphorylated during cytokinesis. Finally, the data show that ectopically expressed Survivin inhibits polyploidization in vascular smooth muscle cells. Hence, aberrant chromosome passenger protein activity and endomitosis are associated with VSMC polyploidization. PMID- 15817154 TI - Expression of Rac1b stimulates NF-kappaB-mediated cell survival and G1/S progression. AB - The small GTPase Rac1 can stimulate various signaling pathways following a tightly controlled GDP-GTP exchange. A splicing variant designated Rac1b was found to exist predominantly in the active GTP-bound state but the functional consequences of its expression remain unknown. Here we used mouse fibroblasts as a model to assess the signaling properties of Rac1b. We show that, in contrast to Rac1, expression of wild-type Rac1b is sufficient to stimulate cyclin D1 accumulation and G1/S progression in these cells. Moreover, expression of wild type Rac1b, but not of wild-type Rac1, dramatically increased cell survival in the presence of only minimal growth stimuli. Both cellular responses were blocked by the NF-kappaB super-repressor IkappaBalpha(A32A36). Active Rac1b induced the phosphorylation and membrane translocation of IkappaBalpha, a prerequisite for the activation of NF-kappaB. These data demonstrate that Rac1b is a highly active Rac1 variant that stimulates cell cycle progression and cell survival in pathways involving NF-kappaB. PMID- 15817155 TI - Survivin enhances telomerase activity via up-regulation of specificity protein 1- and c-Myc-mediated human telomerase reverse transcriptase gene transcription. AB - Suppression of apoptosis is thought to contribute to carcinogenesis. Survivin, a member of the inhibitor-of-apoptosis family, blocks apoptotic signaling activated by various cellular stresses. Since elevated expression of survivin observed in human cancers of varied origin was associated with poor patient survival, survivin has attracted growing attention as a potential target for cancer treatment. Immortalization of cells also is required for carcinogenesis; telomere length maintenance by telomerase is required for cancer cells to proliferate indefinitely. Yet how cancer cells activate telomerase remains unclear. We therefore examined possible interrelationships between survivin expression and telomerase activity. Correlation between survivin and human telomerase reverse transcriptase (hTERT) expression was observed in colon cancer tissues, and overexpression of survivin enhanced telomerase activity by up-regulation of hTERT expression in LS180 human colon cancer cells. DNA-binding activities of specificity protein 1 (Sp1) and c-Myc to the hTERT core promoter were increased in survivin gene transfectant cells. Phosphorylation of Sp1 and c-Myc at serine and threonine residues was enhanced by survivin, while total amounts of these proteins were unchanged. Further, "knockdown" of survivin by a small inhibitory RNA decreased Sp1 and c-Myc phosphorylation. Thus survivin participates not only in inhibition of apoptosis, but also in prolonging cellular lifespan. PMID- 15817156 TI - NF45/ILF2 tissue expression, promoter analysis, and interleukin-2 transactivating function. AB - NF45/ILF2 associates with NF90/ILF3 in the nucleus and regulates IL-2 gene transcription at the antigen receptor response element (ARRE)/NF-AT DNA target sequence (P.N. Kao, L. Chen, G. Brock, J. Ng, A.J. Smith, B. Corthesy, J. Biol. Chem. 269 (1994) 20691-20699). NF45 is widely expressed in normal tissues, especially testis, brain, and kidney, with a predominantly nuclear distribution. NF45 mRNA expression is increased in lymphoma and leukemia cell lines. The human and murine NF45 proteins differ only by substitution of valine by isoleucine at amino acid 142. Fluorescence in situ hybridization localized the human NF45 gene to chromosome 1q21.3, and mouse NF45 gene to chromosome 3F1. Promoter analysis of 2.5 kB of the murine NF45 gene reveals that significant activation is conferred by factors, possible including NF-Y, that bind to the CCAAT-box sequence. The function of human NF45 in regulating IL-2 gene expression was characterized in Jurkat T-cells stably transfected with plasmids directing expression of NF45 cDNA in sense or antisense orientations. NF45 sense expression increased IL-2 luciferase reporter gene activity 120-fold, and IL-2 protein expression 2-fold compared to control cells. NF45 is a highly conserved, regulated transcriptional activator, and one target gene is IL-2. PMID- 15817157 TI - Positive regulation of Fas gene expression by MSSP and abrogation of Fas-mediated apoptosis induction in MSSP-deficient mice. AB - MSSP has been identified as a transcription factor that regulates the c-myc gene. MSSP was later found to positively or negatively regulate a variety of genes, including alpha-smooth actin, MHC class I, MHC class 2 and the thyrotropin receptor. The knockout mice for the Mssp gene developed by us revealed that these mice became partially embryonic lethal due to a low concentration of progesterone at E2.5. In this study, we further analyzed Mssp-knockout mice and found that the expression of the Fas gene was repressed, resulting in abrogation of Fas-mediated induction of apoptosis both in Mssp-knockout mice and primary thymocytes. MSSP was then found to stimulate promoter activity of the Fas gene by binding to a region spanning -1035 to -635 in chromatin immunoprecipitation assays. Binding of MSSP in the MSSP-binding sequence, TCTAAT, located in this region was confirmed by mobility shift assays, and deletion of this sequence from the Fas promoter was found to result in loss of MSSP-dependent stimulating activity. The results suggest that MSSP is an important mediator for Fas-induced apoptosis in vivo and in vitro. PMID- 15817158 TI - Functional characterization of the C. elegans nephrocystins NPHP-1 and NPHP-4 and their role in cilia and male sensory behaviors. AB - Autosomal dominant polycystic kidney disease (ADPKD) and nephronophthisis (NPH) share two common features: cystic kidneys and ciliary localized gene products. Mutation in either the PKD1 or PKD2 gene accounts for 95% of all ADPKD cases. Mutation in one of four genes (NPHP1-4) results in nephronophthisis. The NPHP1, NPHP2, PKD1, and PKD2 protein products (nephrocystin-1, nephrocystin-2 or inversin, polycystin-1, and polycystin-2, respectively) localize to primary cilia of renal epithelia. However, the relationship between the nephrocystins and polycystins, if any, is unknown. In the nematode Caenorhabditis elegans, the LOV 1 and PKD-2 polycystins localize to male-specific sensory cilia and are required for male mating behaviors. To test the hypothesis that ADPKD and NPH cysts arise from a common defect in cilia, we characterized the C. elegans homologs of NPHP1 and NPHP4. C. elegans nphp-1 and nphp-4 are expressed in a subset of sensory neurons. GFP-tagged NPHP-1 and NPHP-4 proteins localize to ciliated sensory endings of dendrites and colocalize with PKD-2 in male-specific sensory cilia. The cilia of nphp-1(ok500) and nphp-4(tm925) mutants are intact. nphp-1; nphp-4 double, but not single, mutant males are response defective. We propose that NPHP 1 and NPHP-4 proteins play important and redundant roles in facilitating ciliary sensory signal transduction. PMID- 15817159 TI - Constitutively active Notch1 induces growth arrest of HPV-positive cervical cancer cells via separate signaling pathways. AB - Notch signaling plays a key role in cell-fate determination and differentiation in different organisms and cell types. Several reports suggest that Notch signaling may be involved in neoplastic transformation. However, in primary keratinocytes, Notch1 can function as a tumor suppressor. Similarly, in HPV positive cervical cancer cells, constitutively active Notch1 signaling was found to cause growth suppression. Activated Notch1 in these cells represses viral E6/E7 expression through AP-1 down-modulation, resulting in increased p53 expression and a block of pRb hyperphosphorylation. Here we show that in cervical cancer cell lines in which Notch1 ability to repress AP-1 activity is impaired, Notch1-enforced expression elicits an alternative pathway leading to growth arrest. Indeed, activated Notch1 signaling suppresses activity of the helix-loop helix transcription factor E47, via ERK1/2 activation, resulting in inhibition of cell cycle progression. Moreover, we found that RBP-Jkappa-dependent Notch signaling is specifically repressed in cervical cancer cells and this repression could provide one such mechanism that needs to be activated for cervical carcinogenesis. Finally, we show that inhibition of endogenous Notch1 signaling, although results in a proliferative advantage, sensitizes cervical cancer cell lines to drug-induced apoptosis. Together, our results provide novel molecular insights into Notch1-dependent growth inhibitory effects, counteracting the transforming potential of HPV. PMID- 15817160 TI - Macaque trophoblast migration is regulated by RANTES. AB - In human and non-human primates, migratory trophoblasts penetrate the uterine epithelium, invade the endometrium, enter the uterine vasculature, and migrate within the arteries. The mechanisms that regulate this directional migration are unknown. We have used early gestation macaque trophoblasts to test the hypothesis that trophoblast migration is regulated by the chemokine, Regulated on Activation T-Cell Expressed and Secreted (RANTES). Immunohistochemical analysis of cryosections of endometrial tissue showed expression of RANTES by stromal cells and vascular cells. Isolated endothelial cells expressed RANTES as determined by immunocytochemistry and RT-PCR analyses. Immunohistochemical analysis of endometrial cryosections showed that the RANTES receptor, CCR5, was expressed by trophoblasts on anchoring villi and by cells within the trophoblastic shell. Cytokeratin-positive/CCR5-positive cells, consistent with trophoblasts, were also found scattered within the stroma and were often clustered around blood vessels. Isolated trophoblast cells expressed CCR5 as determined by immunocytochemistry and RT-PCR analyses. Isolated trophoblasts migrated towards RANTES when cultured in migration chambers and migration was reduced in the presence of anti-CCR5 antibody. When trophoblasts were cultured on dishes coated with recombinant RANTES, expression of beta1 integrin was increased. The RANTES-induced increase in beta1 integrin expression was inhibited by pertussis toxin. These data suggest a role for RANTES and CCR5 in the regulation of trophoblast migration within the endometrium and within the uterine vasculature. PMID- 15817161 TI - Rapid changes in transcription and chromatin structure of ribosomal genes in yeast during growth phase transitions. AB - Transcription of ribosomal genes is coordinated with cellular growth. Changes in transcription may be influenced by an alteration in the number of active ribosomal genes and/or a change in the transcription rate of active genes. We measured changes in rDNA transcription during growth phase transitions in the yeast Saccharomyces cerevisiae and the concomitant changes in chromatin structure of the ribosomal genes. A quantitative transcription run-on (TRO) assay was developed to monitor transcription of ribosomal genes, and rDNA chromatin was separated into active (non-nucleosomal) and inactive (nucleosomal) genes using psoralen photo-crosslinking. TRO indicates that transcription levels of ribosomal genes drop dramatically as cells enter stationary phase, but are rapidly restored when cells are diluted into fresh medium. However, changes in the proportion of active genes during these transitions, although equally rapid, represented only a small fraction of the total rDNA. We conclude that changes in rDNA chromatin structure are temporally coordinated with growth rate, but quantitatively insufficient to account for changes in transcription. These results support the model that regulation of rRNA synthesis occurs mainly by altering the transcription rate of active ribosomal genes, and changes in the number of active rDNA gene copies contribute much less to this regulation. PMID- 15817162 TI - New insights into the nanometer-scaled cell-surface interspace by cell-sensor measurements. AB - The culture of adherent cells on solid surfaces is an established in vitro method, and the adhesion process of a cell is considered as an important trigger for many cellular processes (e.g., polarity and tumor genesis). However, not all of the eliciting biochemical or biophysical reactions are yet understood. Interestingly, there are not much experimental data about the impact that the interspace between an adherent cell and the (solid) substrate has on the cell's behavior. This interspace is mainly built by the basolateral side of epithelial cells and the substrate. This paper gives some new results of non-invasive and non-optical measurements in the interspace. The measurements were made with silicon cell-sensor hybrids. Measurements of acidification, adhesion, and respiration are analyzed in view of the situation in the interspace. The results show that, in general, the release of an ion or molecule on the basolateral side can have much more influence on the biophysical situation than a release of an ion or molecule on the apical side. In particular, the apical acidification (i.e., amount of extruded protons) of, e.g., epithelial tumor cells is several orders of magnitude higher than the basolateral acidification. These experimental results are a simple consequence of the fact that the basolateral volume of the interspace is several orders of magnitudes smaller than the apical volume. These results have the following consequences for the cell adhesion: PMID- 15817163 TI - Single cell analysis of G1 check points-the relationship between the restriction point and phosphorylation of pRb. AB - Single cell analysis allows high resolution investigation of temporal relationships between transition events in G1. It has been suggested that phosphorylation of the retinoblastoma tumor suppressor protein (pRb) is the molecular mechanism behind passage through the restriction point (R). We performed a detailed single cell study of the temporal relationship between R and pRb phosphorylation in human fibroblasts using time lapse video-microscopy combined with immunocytochemistry. Four principally different criteria for pRb phosphorylation were used, namely (i) phosphorylation of residues Ser795 and Ser780, (ii) degree of pRb-association with the nuclear structure, a property that is closely related with pRb phosphorylation status, (iii) release of the transcription factor E2F-1 from pRb, and (iv) accumulation of cyclin E, which is dependent on phosphorylation of pRb. The analyses of individual cells revealed that passage through R preceded phosphorylation of pRb, which occurs in a gradually increasing proportion of cells in late G1. Our data clearly suggest that pRb phosphorylation is not the molecular mechanism behind the passage through R. The restriction point and phosphorylation of pRb thus seem to represent two separate check point in G1. PMID- 15817164 TI - Binding of Par-4 to the actin cytoskeleton is essential for Par-4/Dlk-mediated apoptosis. AB - Prostate apoptosis response-4 (Par-4) is a 38-kDa protein originally identified as a gene product upregulated in prostate cancer cells undergoing apoptosis. Cell death mediated by Par-4 and its interaction partner DAP like kinase (Dlk) is characterized by dramatic changes of the cytoskeleton. To uncover the role of the cytoskeleton in Par-4/Dlk-mediated apoptosis, we analyzed Par-4 for a direct association with cytoskeletal structures. Confocal fluorescence microscopy revealed that endogenous Par-4 is specifically associated with stress fibers in rat fibroblasts. In vitro cosedimentation analyses and in vivo FRET analyses showed that Par-4 directly binds to F-actin. Actin binding is mediated by the N terminal 266 amino acids, but does not require the C-terminal region of Par-4 containing the leucine zipper and the death domain. Furthermore, the interaction of Par-4 with actin filaments leads to the formation of actin bundles in vitro and in vivo. In rat fibroblasts, this microfilament association is essential for the pro-apoptotic function of Par-4, since both disruption of the actin cytoskeleton by cytochalasin D treatment and overexpression of Par-4 constructs impaired in actin binding result in a significant decrease of apoptosis induction by Par-4 and Dlk. We propose a model, in which Par-4 recruits Dlk to stress fibers, leading to enhanced phosphorylation of the regulatory light chain of myosin II (MLC) and to the induction of apoptosis. PMID- 15817165 TI - Direct metabolic regulation of beta-catenin activity by the p85alpha regulatory subunit of phosphoinositide 3-OH kinase. AB - Class IA phosphoinositide 3-OH kinases (PI3K) are lipid kinases composed of catalytic and regulatory subunits. These lipid kinases can regulate the metabolic stability and signaling activity of beta-catenin, a central component of the E cadherin/catenin cell-cell adhesion complex, and of the Wnt signaling pathway. This regulation occurs at the level of glycogen synthase kinase 3 (GSK3), a serine/threonine kinase that marks beta-catenin to enter a destruction pathway. In addition, the regulatory subunit p85alpha directly binds beta-catenin, but the role of this interaction in the context of the lipid kinase regulation of beta catenin signaling is unknown. Here we report that expression of exogenous p85alpha in mouse keratinocytes increases the metabolic stability and has a strong synergistic effect on the transcriptional activity of beta-catenin. Both effects are associated to the formation of beta-catenin/p85alpha and inhibition of beta-catenin/APC complexes and are independent of GSK3 and PI3K activities. These findings suggest that p85alpha can act as a direct metabolic regulator of beta-catenin activity. PMID- 15817166 TI - A secreted form of P-cadherin is expressed in malignant melanoma. AB - Cadherins are Ca-dependent homophilic cell-cell adhesion molecules which are responsible for correct location of cells and tissue integrity. They are critical for development and maintenance of epithelial architecture. Aberrantly expressed cadherins are known to be involved in malignant transformation of different types of tissues. In this study, we show the expression of a short truncated 50 kDa form of the N-terminal part of P-cadherin in seven melanoma cell lines compared to melanocytes and keratinocytes. In vitro protein analysis on cell culture supernatant as well as immunohistochemistry of primary and metastatic melanoma tissue revealed the expression of this short form of P-cadherin. Furthermore, analysis showed that this short 50 kDa form of P-cadherin is secreted by melanoma cells in contrast to the membrane bound form in melanocytes. Analysis on mRNA level detected only exon 1 to 10 of P-cadherin resulting in the 50 kDa form missing the transmembrane and cytoplasmatic region. Genomic sequence analysis did not show any mutations in melanoma cells neither in the exons nor in the exon intron boundaries. Furthermore, there was no loss of exons 11-16 on the genomic level. Functionally, the secreted form of P-cadherin could play a role as regulator of the homophilic interaction between P-cadherin molecules by antagonizing their biological role acting as a dominant negative form to interrupt cell-cell attachment. PMID- 15817167 TI - Proteolysis leads to the appearance of the long form of beta3-endonexin in human platelets. AB - After vessel injury, platelets adhere to the subendothelial matrix. Platelet adhesion leads to activation of the platelet integrin alpha(IIb)beta3, which then binds to fibrinogen, leading to platelet aggregation. It has been shown that a beta3-integrin binding protein, beta3-endonexin, can activate the integrin alpha(IIb)beta3 expressed in transfected CHO cells. Several isoforms of beta3 endonexin are known but it is not clear which isoforms are expressed in platelets and what role they may play during haemostasis. Here, we show that the long form of beta3-endonexin (EN-L) can be detected in platelet lysates several hours after thrombus formation, after long-term storage of platelets and after glucose deprivation. After subcellular fractionation, EN-L is found in the detergent insoluble fraction suggesting that it might be associated with the cytoskeleton. EN-L generation is temperature and Ca++ dependent and requires physiological salt concentrations. Proteolysis is responsible for the appearance of EN-L since a calpain inhibitor prevents its formation and the addition of calpain to platelet lysates induces its formation. The appearance of EN-L seems to be linked to apoptotic events occurring during long-term storage of platelets and, possibly, during late steps of haemostasis after thrombus formation. PMID- 15817168 TI - Changes in memory for emotional material in amyotrophic lateral sclerosis (ALS). AB - The current study sought to examine the performance of non-demented ALS patients on neuropsychological tests involving emotional perception and memory. Nineteen non-demented patients with ALS were compared with 20 healthy controls on assessments of facial expression recognition, social judgement ratings of faces and recognition memory for emotional words. Patients and controls were well matched to exclude a range of potentially confounding variables. The patients and controls demonstrated significant differences on only one test of cognitive functioning. The ALS group demonstrated a failure to show the normative pattern of enhanced recognition memory for emotional words compared to neutral words and produced higher scores than controls on recognition memory for neutral words. These findings suggest that patients with ALS show a different pattern of cognitive performance with respect to memory for emotional material when compared to healthy adults. PMID- 15817169 TI - Brain regions associated with successful and unsuccessful retrieval of verbal episodic memory as revealed by divided attention. AB - Which brain regions are implicated when words are retrieved under divided attention, and what does this tell us about attentional and memory processes needed for retrieval? To address these questions we used fMRI to examine brain regions associated with auditory recognition performed under full and divided attention (DA). We asked young adults to encode words presented auditorily under full attention (FA), and following this, asked them to recognize studied words while in the scanner. Attention was divided at retrieval by asking participants to perform either an animacy task to words, or odd-digit identification task to numbers presented visually, concurrently with the recognition task. Retrieval was disrupted significantly by the word-, but not number-based concurrent task. A corresponding decrease in brain activity was observed in right hippocampus, bilateral parietal cortex, and left precuneus, thus demonstrating, for the first time, involvement of these regions in recognition under DA at retrieval. Increases in activation of left prefrontal cortex (PFC), associated with phonological processing, were observed in the word- compared to number-based DA condition. Results suggest that the medial temporal lobe (MTL) and neo-cortical components of retrieval, believed to form the basis of episodic memory traces, are disrupted when phonological processing regions in left PFC are engaged simultaneously by another task. Results also support a component-process model of retrieval which posits that MTL-mediated retrieval does not compete for general cognitive resources but does compete for specific structural representations. PMID- 15817170 TI - Evaluating a split fovea model of visual word recognition: effects of case alternation in the two visual fields and in the left and right halves of words presented at the fovea. AB - Two experiments are reported exploring the effect of cAsE aLtErNaTiOn on lexical decisions to words and nonwords presented laterally or centrally. In line with previous research, Experiment 1 found that case alternation slowed lexical decision responses to words more in the right visual field (RVF) than in the left visual field (LVF). In Experiment 2, the words and nonwords were all presented centrally. There were three conditions, a condition in which the word and nonwords were presented in lower case letters, a condition in which the letters to the left of the central fixation were case alternated (e.g., aMbItion, mOdLants) and a condition in which the letters to the right of fixation were case alternated (e.g., collApSe, pireNtOl). Alternating the case of letters to the right of fixation slowed lexical decision responses more than alternating letter case to the left of fixation. The results provide further support for a split fovea account of visual word recognition according to which those letters of a centrally-fixated word that fall to the left of fixation are processed initially by the right cerebral hemisphere while those letters that fall to the right of fixation are processed initially by the left cerebral hemisphere, with the characteristics of the left and right hemispheres being revealed in the processing of initial and final letters in centrally presented words. PMID- 15817171 TI - Representational neglect and navigation in real space. AB - Topographical orientation relies on several cognitive strategies adopted by humans to move within the environment. In the present study, we investigate whether mental representation disorders affect specific cognitive mechanisms subserving human orientation. In order to differentiate distinct cognitive mechanisms involved in topographical orientation, we created a human version of the well-known "Morris Water Maze" and tested left and right brain damaged patients in a place-learning task. The test required the subjects to explore an experimental room in which no visual cues were present, find a target location, and then reach it in different conditions. The ability to memorize target locations in short- and long-term memory was also assessed. We found that all participants were able to reach the target location by using idiothetic cues (vestibular inputs, motor efferent copy, etc.). On the other hand, when starting position changed and re-orientation was necessary to reach the target location, in order to compute a new trajectory, only patients affected by representational neglect got lost. These results provide the first neuropsychological evidence of involvement of mental representation in a specific cognitive process allowing humans to reach a target place from any location in the environment. PMID- 15817172 TI - Action (verb) generation in HIV-1 infection. AB - It has been proposed that verb generation is primarily associated with left fronto-basal ganglia circuits, whereas the generation of nouns is principally mediated by dominant left temporo-parietal networks. Consistent with this premise, action (verb) fluency - a verbal fluency task requiring the spontaneous generation of verbs - has shown greater sensitivity to frontal-basal ganglia pathophysiology (e.g., dementia in Parkinson's disease (PDD)) than noun fluency. The present study examined action and noun fluency in persons with HIV-1 infection-a disease known to be associated with a frontal-basal ganglia circuit neuropathogenesis. Action and noun ("animals") verbal fluency protocols were administered to 97 persons with HIV-1 infection and 20 demographically comparable healthy comparison (HC) subjects. A significant interaction emerged between verbal fluency task and HIV-1 serostatus such that the HIV+ group generated significantly fewer actions (verbs) relative to the HC sample. Findings indicate that persons infected with HIV-1 experience difficulty rapidly generating verbs, but not nouns from semantic memory. Considering the prominent frontal-basal ganglia circuit neuropathophysiology of HIV-1 infection, these data are consistent with the hypothesized dissociation between noun and verb generation as pertains to generative fluency. PMID- 15817173 TI - Electrophysiological indices of strategic episodic retrieval processing. AB - Event-related potentials (ERPs) were acquired during test phases of a recognition memory exclusion task, in order to contribute to current understanding of the processes responsible for the ways in which memory retrieval can be controlled strategically. Participants were asked to endorse old words from one study task (targets) and to reject new test words as well as those from a second study task (non-targets). The study task designated as the target category varied across test phases. The left-parietal ERP old/new effect--the electrophysiological signature of recollection--was reliable for targets only in all test phases, consistent with the view that participants control recollection strategically in service of task demands. The contrast between the ERPs evoked by new test words separated according to target designation revealed reliable differences at midline, anterior and right-hemisphere locations. These differences likely reflect processes that form part of a retrieval attempt and are interpreted here as indices of processes that are important for the strategic regulation of episodic retrieval. PMID- 15817174 TI - Memory for time intervals is impaired in left hemi-Parkinson patients. AB - The basal ganglia have been proposed as one of the neural correlates of timekeeping functions. Both encoding and memory retrieval components for time perception are impaired in Parkinson's disease (PD). The aim of our study was to investigate in hemi-Parkinsonian patients the existence of a specific alteration in memory for time depending on the affected side, to better understand the contribution of the left or right basal ganglia circuits in different components of time perception. Right and left hemi-PD patients performed a time reproduction task in which they were required to reproduce in the same session short (5 s) and long (15 s) time intervals, in off- and on-therapy condition. While the right hemi-PD patients overestimated the shorter interval, only the left hemi-PD group showed the memory migration effect, overestimating the shorter and underestimating the longer time intervals. These results argue for a critical involvement of the right basal ganglia in memory retrieval for time intervals, in the range of seconds. PMID- 15817175 TI - Dissociation of conscious and unconscious repetition priming effect on event related potentials. AB - Although the neural basis of the unconscious priming effect has previously been investigated, the results of these studies have possibly been contaminated by a conscious priming effect. The aim of the present study was to dissociate the effects of conscious and unconscious priming on event-related potential (ERP) by using the process-dissociation procedure. A prime word was presented briefly, followed by a word-stem, in each trial. Under the inclusion condition, subjects were instructed to complete the word-stem using the prime word, while under the exclusion condition subjects were asked to complete the word-stem with a word not seen as prime. The behavioral priming effect was obtained under both conditions, indicating that the prime words were processed unconsciously and influenced the word-stem completion task. We found that two ERP components were affected by repetition priming. First, the N400 amplitude was decreased by word repetition under the inclusion condition, but not under the exclusion condition. This result suggests that N400 would reflect conscious lexical processing, but not unconscious lexical activation. Second, the negativity at left front lateral region was enhanced by word repetition under the exclusion condition. We discuss this finding herein in relation to the activity of the left inferior prefrontal cortex with regard to word semantic processing. PMID- 15817176 TI - "I can write seven but I can't say it": a case of domain-specific phonological output deficit for numbers. AB - While it is established that numbers represent a category-specific semantic domain being dissociable from other semantic domains, it is a debated issue whether the category-specific dissociability persists at a level of lexical processing or whether words and number words are processed by the same lexical mechanisms. In most of the documented cases of brain damage, lexical processing of words and number words was equally spared or impaired. Here, we describe the case of a patient who after recovering from his naming disorders, showed a selective deficit in the spoken production of numbers with spared number writing and number comprehension abilities. After having ruled out alternative interpretations, we can conclude that these results indicate that words and numbers are not only categorically organised at the semantic level but that they are also processed by different lexical mechanisms. In addition, the results argue in favour of the existence of different modality-specific output lexicons for the written and spoken production of numerals. PMID- 15817177 TI - Attentional control parameters following parietal-lobe damage: evidence from normal subjects. AB - Attentional control involves the factors, or cognitive parameters, that determine which environmental inputs receive attention and which do not. Cognitive studies of attentional control have highlighted two general classes of control parameters, bottom-up (data driven or exogenous) parameters and top-down (goal driven or endogenous) parameters. Which of these control parameters is affected following parietal-lobe damage? In parietal-damaged patients, it is possible that a disorder in one control parameter (e.g. goal driven) would appear as a disorder in another parameter (e.g. data driven). To investigate the control parameters that might be affected in parietal patients, we simulated neglect in normal participants by disrupting data-driven information processing. When half of a computer monitor was degraded by translucent tracing paper while normal participants performed a cued spatial attention task (Experiment 1), the normal participants showed a pattern of results similar to patients with unilateral parietal-lobe damage--the so-called "disengage deficit." This pattern of results replicated when neutral attentional cues were included in the experiment (Experiment 2). However, the disengage deficit was not simulated in normal participants with predictive central symbolic cues (Experiment 3) or predictive peripheral cues (Experiment 4). Because perceptual degradation influences data driven attentional control parameters, we suggest that these control parameters may be disrupted following parietal-lobe damage. PMID- 15817178 TI - Brain-based mechanisms underlying complex causal thinking. AB - We use functional magnetic resonance imaging (fMRI) and behavioral analyses to study the neural roots of biases in causal reasoning. Fourteen participants were given a task requiring them to interpret data relative to plausible and implausible causal theories. Encountering covariation-based data during the evaluation of a plausible theory as opposed to an implausible theory selectively recruited neural tissue in the prefrontal and occipital cortices. In addition, the plausibility of a causal theory modulated the recruitment of distinct neural tissue depending on the extent to which the data were consistent versus inconsistent with the theory provided. Specifically, evaluation of data consistent with a plausible causal theory recruited neural tissue in the parahippocampal gyrus, whereas evaluating data inconsistent with a plausible theory recruited neural tissue in the anterior cingulate, left dorsolateral prefrontal cortex, and precuneus. We suggest that these findings provide a neural instantiation of the mechanisms by which working hypotheses and evidence are integrated in the brain. PMID- 15817180 TI - A 3-year follow-up study of 'orientation agnosia'. AB - Dissociation between the ability to recognize misoriented objects and to determine their orientation has been reported in a small number of patients, but the long-term course of this deficit has not been reported so far. Here, we describe the case of a 32-year-old female who had bilateral occipito-temporal damage caused by a cerebrovascular accident. Neuropsychological assessment performed at 6 months after the occurrence of the cerebrovascular accident revealed that she was almost generally agnostic for object orientation. The patient was then re-tested 3 years later, when she showed apparently striking recovery in her ability to determine object orientation. However, closer examination revealed that she still displayed the same impairment, although at this time, it was only for objects presented in non-cardinal angles. Moreover, she had problems mostly discriminating orientations that differed by small amounts. The ability of patients to discriminate a variety of orientations should be further tested in future investigations in this field. PMID- 15817179 TI - Dissociable elements of human foresight: a role for the ventromedial frontal lobes in framing the future, but not in discounting future rewards. AB - Impaired future thinking may be a core aspect of impulsive decision making. Recent efforts to understand the brain processes that underlie impulsivity have suggested a role for the frontal lobes. However, future thinking is unlikely to be a unitary process, and the frontal lobes are not a homogeneous entity. The present study contrasted the effects of dorsolateral and ventromedial frontal lobe damage on two distinct aspects of future thinking in humans. Temporal discounting, the subjective devaluation of reward as a function of delay, is not affected by frontal lobe injury. In contrast, a normal future time perspective (a measure of the length of an individual's self-defined future) depends on the ventromedial, but not dorsolateral, frontal lobes. Furthermore, investigation of the relationship of these two measures with classical symptoms of frontal lobe damage indicates that future time perspective correlates with apathy, not impulsivity. Apathy may deserve more attention in understanding both impaired future thinking and the impaired decision making that may result. PMID- 15817181 TI - Allothetic orientation and sequential ordering of places is impaired in early stages of Alzheimer's disease: corresponding results in real space tests and computer tests. AB - Spatial disorientation and learning problems belong to the integral symptoms of Alzheimer's disease (AD). A circular arena for human subjects (2.9 m diameter, 3 m high) was equipped with a computerized tracking system, similar to that used in animals. We studied navigation in 11 subjects diagnosed with early stages of Alzheimer's disease (AD), 27 subjects with subjective problems with memory or concentration, and 10 controls. The task was to locate one or several unmarked goals using the arena geometry, starting position and/or cues on the arena wall. Navigation in a real version and a computer map view version of the tests yielded similar results. The AD group was severely impaired relative to controls in navigation to one hidden goal in eight rotated positions. The impairment was largest when only the cues on the wall could be used for orientation. Also, the AD group recalled worse than controls the order of six sequentially presented locations, though they recalled similarly to controls the positions of the locations. The group with subjective problems was not impaired in any of the tests. Our results document the spatial navigation and non-verbal episodic memory impairment in the AD. Similar results in real and map view computer tests support the use of computer tests in diagnosis of cognitive disturbances. PMID- 15817182 TI - Spatial orientation strategies in Morris-type virtual water task for humans. AB - The present study characterized frequent motion patterns (search strategies) that occurred during spatial navigation in a virtual maze. The research focused on identifying and characterizing some search strategies, the temporal progression of strategy-use, and their role in spatial performance. Participants were 112 undergraduate students (42 males and 70 females). We identified three search strategies that predicted spatial performance. Enfilading refers to an approach withdrawal pattern of active exploration near a target location. Thigmotaxis refers to a search strategy that involves continuous contact with the circular wall of the maze. Visual scan involves active visual exploration while the subject remains in a fixed spatial location and turns round. In addition to identifying these motion patterns, some significant points of the spatial learning process were also detailed where strategies appeared to shift systematically. The applied search strategies in these transitional points have determined overall spatial performance. PMID- 15817183 TI - Blockade of adenosine A2A receptors reverses short-term social memory impairments in spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) exhibit impairment across several cognitive domains such as attention, short-term memory and spatial reference memory. These cognitive deficits have been variously attributed to disrupted dopaminergic, cholinergic and adenosinergic neurotransmitter function. However, social memory in SHR has not been investigated. In the present study, we therefore evaluated whether SHR exhibit altered short-term social memory abilities compared to normotensive Wistar rats (WIS) through two experimental paradigms (social recognition and habituation-dishabituation tests). We also compared the performance of SHR and WIS rats in the object recognition test. SHR exhibited significantly impaired performance in both models of social memory, but not in the object recognition test, demonstrating a selective deficit in the ability to recognize a juvenile rat after a short period of time. The administration of acute doses of the non-selective adenosine receptor antagonist caffeine (3.0 or 10.0 mg/kg, i.p.) and the adenosine A2A receptor 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, 0.5 or 1.0 mg/kg, i.p.) but not the adenosine A1 receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX, 1.0 or 3.0 mg/kg, i.p.) reversed this social memory impairment in SHR, but these treatments did not alter the hypertension state. These results demonstrate an impairment of short-term social memory in SHR and the involvement of the adenosine A2A receptors in this alteration. PMID- 15817184 TI - Early environmental experience alters baseline and ethanol-induced cognitive impulsivity: relationship to forebrain 5-HT1A receptor binding. AB - The relationship between impulsivity and drug abuse is poorly understood despite evidence that impulsive behaviour both predicts, and is a consequence of, drug use. Moreover, although there are clear individual differences in the propensity to addiction, this relationship has not been investigated with respect to impulsive behaviour. We tested whether early environmental experience would influence behavioural measures of impulsivity, and further, whether this experience would alter impulsive choice following ethanol intoxication. Thirty six male, Long-Evans rats were reared in either isolated (1 rat/cage), standard (2 rats/cage), or enriched (group housed with toys) conditions. After a 3-month rearing period, animals were tested in two operant tasks measuring either motor (go/no-go) or cognitive (delay-to-reinforcement) impulsivity. Rats were then re tested following 0, 0.3, 0.6, 0.9, and 1.2 g/kg ethanol. Forebrain 5-HT1A binding was assessed post-mortem using in vitro receptor autoradiography with the agonist [3H]8-OH-DPAT (3H-8-hydroxy-2-[di-n-propylamino]tetralin). Rearing condition did not influence baseline motor impulsivity, but isolation rearing led to decreased baseline cognitive impulsivity. Ethanol did not affect motor impulsivity, but dose-dependently increased impulsive choice in the delay-to-reinforcement task. Enriched rats were more impulsive overall, and isolation-reared rats only showed a shift in impulsive behaviour after 1.2 g/kg. Isolation rearing decreased, and enrichment rearing increased 5-HT1A binding in the frontal pole of the cortex following experience in the delay-to-reinforcement task. Isolation-reared rats also showed a significant decrease in 5-HT1A binding in the dentate gyrus of the ventral hippocampus following experience in the delay-to-reinforcement relative to the go/no-go task. These data indicate that differential rearing has a significant influence on cognitive impulsivity, and that altered serotonergic function may underlie these differences. PMID- 15817185 TI - Conditioned suppression and freezing as measures of aversive Pavlovian conditioning: effects of discrete amygdala lesions and overtraining. AB - Freezing and suppression are measures of conditioned fear that correlate in unlesioned animals. Both the basolateral (BLA) and central (CeN) nuclei of the amygdala are required for conditioned freezing, though there can be recovery with overtraining. The neuroanatomical substrates of conditioned suppression are less clear, with evidence both for a specific requirement of the CeN and for disruption by BLA lesions. The present study investigated the impact of selective excitotoxic lesions of the BLA and CeN upon the acquisition and expression of conditioned fear, measured by freezing and both on-baseline and off-baseline conditioned suppression in the same rats. BLA and CeN lesions both abolished all measures of conditioned fear after 9 trials of fear conditioning. However, when conditioning was extended to 33 trials, whereas rats with combined lesions of both the BLA and CeN continued to show no conditioned fear responses, there was a pattern of recovery observed after selective lesions. There was a partial recovery of freezing with both lesions, and full recovery of conditioned suppression, except for off-baseline suppression in CeN lesioned rats. These results indicate that with few conditioning trials, both the BLA and CeN are required in a serial manner for conditioned fear responses, but that overtraining can mitigate such impairments, likely involving parallel pathways in and through the amygdala. PMID- 15817186 TI - Piracetam counteracts the effects of amitriptyline on inhibitory avoidance in CD1 mice. AB - The purpose of the present work was to study the effects of amitriptyline on animal cognition in relation to some characteristics of its therapeutic effects. The modulation of acute and chronic effects of amitriptyline on inhibitory avoidance in male and female mice by piracetam was investigated. In Experiment 1, mice were subjected to the training phase of inhibitory avoidance conditioning 60 min after acute piracetam (100 mg/kg) or physiological saline administration. Immediately after the behavioural task, they received a single injection of the tricyclic antidepressant amitriptyline (30 mg/kg) or physiological saline. Twenty four hours later, subjects were tested for avoidance. In Experiment 2, the same doses of amitriptyline and piracetam were chronically administered. Mice were subjected to the training phase of inhibitory avoidance on the 22nd day, and to the test phase 24 h later. Forty-five minutes after test, subjects explored the elevated plus-maze for 5 min in order to assess whether the effects of amitriptyline on avoidance performance may reflect general behavioural changes. Results obtained were that: (a) acute and chronic amitriptyline impaired inhibitory avoidance of male and female mice, (b) piracetam counteracted the effect of acutely administered amitriptyline on inhibitory avoidance, and (c) piracetam counteracted the effects of chronically administered amitriptyline in males but not females in the same learning task. These effects do not seem to be mediated by non-specific drug effects on spontaneous motor activity or anxiety. PMID- 15817187 TI - Familiarity increases consistency in animal tests. AB - This experiment investigated the effect of identity of the experimenter and familiarity to their test animals on results obtained from a standard test of anxiety. We found that having different experimenters perform the same test (i.e. elevated plus maze) using the same equipment and rats from the same breeding colony within the same room of the same laboratory significantly affects the results if the experimenters are unfamiliar to the animals, but not if they are familiar to them. Familiarity of the test animals with their experimenters should therefore increase consistency in results from animal tests. PMID- 15817188 TI - Detailed analysis of the behavior of Lister and Wistar rats in anxiety, object recognition and object location tasks. AB - The present study, examines some issues in the measure and analysis of behavior in animals. Two strains of rats of both genders were used to illustrate and discuss these issues. We examined to what extent various behavioral measures reflect different or identical emotional or cognitive factors and, how sensitive are the various parameters of a task to differences between strains and genders. Wistar and Lister males and females rats were tested in an anxiety test then in the object recognition task followed by the object location task. Taking advantage of a simple computer program it is possible to: (1) record several parameters of theses tasks and examine the pattern of animal responses toward novelty and/or familiarity; (2) examine whether different measurements of the same response would reflect anxiety response to novelty and, can they discriminate between novelty and familiarity responses to objects; and (3) examine if changes in the pattern of animal responses are reflected by these measurements and, whether anxiety or discrimination is evident mainly during the first minute of the test. The results on the anxiety test show that different measures of the same response proved concordant and revealed significant differences between Lister males and Wistar males. Lister males approached more frequently an object and spent more time on an object in each approach compared to Wistar males in the first 5 min of test and in the total 10 min. They have also shorter latencies between approaches compared to Wistar males. The examination of performance over different time bins was significant with the measure of frequency. Lister male rats approached less frequently the object in the last 5 min of the test compared to the first 5 min. Their performance, however, did not differ from that of the other groups in this last 5 min. In the memory tasks, the measure of the frequency of approaches suggests that Lister male rats were able to discriminate between novel and familiar objects and, between novel and familiar location of objects. The measure of latency of first approach shows that Wistar female rats were able to discriminate between objects only in the spatial memory test. Discrimination in the object recognition task was observed in the first and second minute, and in the total 3 min sessions. Discrimination in the object location task was observed with the measure of frequency of approaches, in the first minute, and in the total 3 min sessions. Results from the total 3 min sessions were more concordant between the different measures of discrimination than results from separate 1 min bins. The results from the two memory tasks show that novelty prevented habituation to re-exposure to the testing environment. In many cases, novelty increased exploration of the objects in the choice phase compared to the sample phase. However, this lack of habituation or increased exploration in the choice phase is not concordant with most results of discrimination between novelty and familiarity from the same type of measurements. PMID- 15817189 TI - Learned irrelevance is disrupted in first-episode but not chronic schizophrenia patients. AB - Learned irrelevance (LIrr) is a pre-exposure effect in which uncorrelated presentations of a conditioned stimulus (CS) and an unconditioned stimulus (US) retard subsequent CS-US association. LIrr is closely related to the phenomenon of latent inhibition (LI). LI refers to the retarding effects of inconsequential stimulus pre-exposure on subsequent conditioning to that stimulus, and is considered to reflect the organism's capacity to ignore irrelevant stimuli. LI is disrupted in schizophrenia patients, due to faster learning of the association between the preexposed CS and the US. A new within-subject target-recognition LIrr procedure was applied. The target was either cued by a priming signal or appeared at random, and priming signals were novel or preexposed cues. Schizophrenia patients were compared to age- and sex-matched control subjects. Normal subjects (n = 24) have shown robust LIrr, namely, faster cue-target associations of novel compared to preexposed cues. Schizophrenia patients at the early stages of their first episode (n = 7) showed LIrr disruption, namely, cue target associations to preexposed cues were as fast as for novel cues. Chronic patients during an acute phase (n = 18) did not show LIrr as they failed to learn the cue-target association. In addition to the LIrr paradigm the same subjects were tested in a covert-orientation task. No differences were observed between the groups on this task. The possible advantages of the new LIrr paradigm are discussed. PMID- 15817190 TI - Persistent activation of select forebrain regions in aggressive, adolescent cocaine-treated hamsters. AB - Hamsters repeatedly exposed to cocaine throughout adolescence display highly escalated offensive aggression compared to saline-treated littermates. The current study investigated whether adolescent cocaine exposure activated neurons in areas of hamster forebrain implicated in aggressive behavior by examining the expression of FOS, i.e., the protein product of the immediate early gene c-fos shown to be a reliably sensitive marker of neuronal activation. Adolescent cocaine-treated hamsters and saline-treated littermates were scored for offensive aggression and then sacrificed 1 day later and examined for the number of FOS immunoreactive (FOS-ir) cells in regions of the hamster forebrain important for aggression control. When compared with non-aggressive, saline-treated controls, aggressive cocaine-treated hamsters showed persistent increases in the number of FOS-ir cells in several aggression regions, including the anterior hypothalamus, nucleus circularis, lateral hypothalamus (i.e., the hypothalamic attack area), lateral septum, and medial and corticomedial amygdaloid nuclei. Conversely, aggressive cocaine-treated hamsters showed a significant decrease in FOS-ir cells in the medial supraoptic nucleus, bed nucleus of the stria terminalis, and central amygdala when compared with controls. However, no differences in FOS-ir cells were found in other areas implicated in aggression such as the paraventricular hypothalamic nucleus, or in a number of non-aggression areas. These results suggest that adolescent cocaine exposure may constitutively activate neurons in select forebrain areas critical for the regulation of aggression in hamsters. A model for how persistent activation of neurons in one of these brain regions (i.e., the hypothalamus) may facilitate the development of the aggressive phenotype in adolescent cocaine-exposed animals is presented. PMID- 15817191 TI - The 5-HT1A receptor and recognition memory. Possible modulation of its behavioral effects by the nitrergic system. AB - Functional activation of the 5-HT1A receptor inhibits cognition, although discrepant findings have also been reported. The present study was designed to investigate the role of the 5-HT1A receptor on recognition memory in the rat. For this purpose, the effects induced by the 5-HT1A agonist R-(+)-8-hydroxy-2-(di-n propylamino)tetralin hydrobromide (8-OH-DPAT) and the 5-HT1A antagonist WAY 100635 on memory were evaluated by using the object recognition task. In addition, the possible involvement of the nitrergic system on 5-HT1A receptor's effects was also assessed by using the same behavioral procedure. In the first dose-response study, post-training administration of 8-OH-DPAT (0.1 and 0.3 mg/kg, subcutaneously (s.c.)) dose-dependently impaired animals' performance in this test. WAY 100635 (0.3 and 1 mg/kg, intraperitoneally (i.p.)) successfully antagonized these 8-OH-DPAT-induced performance deficits. The NO donor molsidomine (2 and 4 mg/kg, i.p.) counteracted cognition deficits produced by the highest dose of 8-OH-DPAT (0.3 mg/kg). Our findings indicate (a) that the 5-HT1A receptor is involved in recognition memory, and (b) that a NO component modulates the effects of the 5-HT1A receptor on learning and memory. PMID- 15817192 TI - Effects of histamine H3 receptor antagonists in two models of spatial learning. AB - Despite the well-described attention and short-term memory enhancing effects of H3 receptor antagonists, and evidence to suggest a close relationship between central histaminergic and cholinergic systems, there is a paucity of evidence for a role for H3 receptor blockade in spatial learning. To address this, we investigated two H3 receptor antagonists in a visual discrimination water maze in rats, and in a Barnes circular maze in mice. Thioperamide and ciproxifan significantly attenuated a scopolamine-induced deficit in the water maze task, while only ciproxifan showed a modest attenuation in the Barnes maze. Taken together, these data suggest a role for H3 receptors in spatial learning that appears to be task-dependent. PMID- 15817193 TI - Differences in affective behaviors and hippocampal allopregnanolone levels in adult rats of lines selectively bred for infantile vocalizations. AB - Allopregnanolone, 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP), a progesterone metabolite, is an endogenous neurosteroid mediating affective behaviors via its positive modulation of GABA(A) receptors. In order to better understand the role of this neurosteroid in individual differences in affective behavior, we used an animal model based on selective breeding for an infantile affective trait, ultrasonic vocalizations (USV). Adult male and female (in either proestrus or diestrus) rats that had been bred for low (low line) or high (high line) rates of USV after maternal separation were tested in a series of affective behavioral tests: open field, emergence, social interaction, defensive freezing, and the Porsolt forced swim task. Concentrations of allopregnanolone in combined hippocampus and amygdala tissue were then measured. low line subjects showed significantly lower anxiety and depression responses in the emergence, open field, and Porsolt forced swim tasks than did high line subjects. Proestrus females exhibited less affective behaviors than diestrus females or males. Allopregnanolone levels in hippocampus/amygdala were significantly higher in low line subjects compared to high line subjects, and in proestrus females compared to diestrus females and males. These data indicate that: (1) affective behaviors in lines selectively bred for an infantile anxiety trait exhibit selection persistence into adulthood; and (2) levels of allopregnanolone in the limbic system parallel selected disparities in affective behavior, suggesting a selection for alterations in the neurosteroid/GABA(A) receptor system in these lines. PMID- 15817194 TI - Implicit learning in a serial choice visual discrimination task in the operant 9 hole box by intact and striatal lesioned mice. AB - Within a broader programme developing murine models of Huntington's disease (HD), we have sought to develop a test of implicit learning for the mouse. Mice were trained in a novel serial visual discrimination task in the '9-hole box' operant test apparatus, followed by retesting after either bilateral quinolinic acid striatal lesions or sham lesions. In the task, each trial involves two sequential responses: an initial light stimulus is presented randomly in one of five holes, to which a nose-poke response results in the first light being extinguished and a second light is illuminated in a different hole. Response to the second light results in food reward, followed by a brief interval before the next trial. When the first light was in one of three of the five holes, the location of the second light was unpredictable in any of the remaining four holes; by contrast, if the first light occurred in one of the other two of the five holes, then the location of the second light was entirely predictable, being the hole two steps to the left or to the right, respectively. Reaction times and accuracy of responding were recorded to both stimuli. The mice learned the task with a degree of accuracy, and they demonstrated clear implicit learning, as measured by increased accuracy and reduced latency to respond to the presentation of the predictable stimulus. Striatal lesions disrupted performance, reducing accuracy for both the first and second stimuli and increasing response latencies for the second stimuli. The decrease in accuracy by the lesioned animals was accompanied by increases in perseverative nose-poking and inappropriate magazine entries throughout the trials, but the lesioned mice still showed a similar benefit (albeit, against a lower baseline of performance) from the implicit knowledge provided on predictable trials. The data validates the task as a sensitive probe for determining implicit learning deficits in the mouse, and suggests that the consequences of striatal lesions, while disrupting performance of skilled stimulus-response habits, are not selective to the process underlying implicit learning. PMID- 15817195 TI - Attention-deficit and hyperactive neurobehavioural characteristics induced by perinatal hypothyroidism in rats. AB - Thyroid hormone is essential for the proper development of the mammalian central nervous system (CNS). In the present study, we examined behavioural alterations caused by transient perinatal hypothyroidism induced by an anti-thyroid drug, propylthiouracil (PTU). This drug produces perinatal disruption of the thyroid system and subsequent behavioural changes, which we investigated using a series of behavioural tests and focusing particularly on attention-deficit/hyperactivity disorder (ADHD)-like behaviours. In the open field test, both male and female rats that had experienced perinatal hypothyroidism (HT rats) showed an increased percent of locomotion behaviour and reduced grooming behaviour, suggesting that HT rats may be hyperactive and show fewer anxiety characteristics. Neither male nor female HT rats showed retention in the passive avoidance test. Male HT rats showed a significantly lower rate of correct avoidance responses than control rats in earlier sessions in the active avoidance test. In addition, we observed significant increases in the number of times that rats crossed the partition during inter-trial intervals and the percent of failure of avoidance during 5 s electrical stimuli in HT rats, suggesting that HT rats are restless, have a shortened attention span and panic easily. In measuring spontaneous motor activity during a period of darkness, male HT rats appeared to plunge into active phase with short, quick steps, while male control rats showed only long active phases during a stress-free period of darkness. These abnormal behavioural characteristics in HT rats might coincide with those found in some cases of ADHD. PMID- 15817196 TI - Epidemiology of Eimeria infections in an Austrian milking sheep flock and control with diclazuril. AB - In a flock of milking sheep from Styria (Austria), the dynamics of Eimeria oocyst excretion was monitored in the lambing season 2003-2004 and a treatment trial with 1 mg/kg diclazuril (single dose on day 0 of study) was conducted in two groups of early weaned lambs. Adult animals (n = 30 ewes, 30 yearlings) excreted oocysts of different species (weekly prevalences from week -7 ante partum to week 5 post-partum: ewes 20-60%, yearlings 38-73%) in low intensities (< or = 6000 oocyst per gram of faeces, highest values in weeks -1 and -4) without clinical signs. Ewes excreted significantly fewer oocysts than yearlings. Lambs in the first group (n = 32 treated animals, 32 controls) excreted oocysts from the seventh day of sampling (average age on day 0: 28 days), those in the consecutive group (n = 32 treated animals, 30 controls) already excreted oocysts on day 0 (average age: 34 days). Treatment resulted in significant reduction of the overall excretion rates on days 7-21 of study compared with untreated controls. Similarly, the excretion intensities were significantly reduced in the treated groups. Overall excretion rates of the pathogenic species (Eimeria ovinoidalis, Eimeria bakuensis, Eimeria weybridgensis/Eimeria crandallis, Eimeria ahsata) were significantly reduced after treatment. Soft faeces and anal soiling in the lambs were significantly reduced and weight gain was increased. A single treatment of lambs with diclazuril before or shortly after the onset of oocyst shedding was sufficient to control oocyst excretion and improve animal health. PMID- 15817197 TI - Immunization protocols against Cryptosporidium parvum in ovines: protection in suckling lambs. AB - Ovine colostrum and milk from immunized ewes were tested for their ability to prevent cryptosporidiosis in the lambs experimentally infected with 10(6) oocysts of Cryptosporidium parvum at 36-48 h of age (day 0 post-infection). All lambs became infected and developed clinical cryptosporidiosis. However, lambs fed by immunized dams have shown shedding involved, significantly, fewer oocysts and lasted for a shorter period than in control lambs. In addition, diarrhoea was less severe. The best results emerged in lambs of ewes immunized by intramuscular injection of an emulsion of 2 ml of Freund's complete adjuvant and 2 ml of C. parvum antigen in sterile phosphate buffered saline solution, administrated four weeks before parturition, together with an intramammary infusion of 25 microg of antigen in 2 ml of sterile PBS emulsified in 2 ml of Freund's incomplete adjuvant, which showed the highest anti-C. parvum titres in lacteal secretions. In their case, the onset of output of oocysts was delayed by two days, the patent period was shortened by three days, their diarrhoea continued for only three days, and the quantity of oocysts shed decreased by 77%. The outcome was that at the end of the study they had a live weight gain of 2 kg more than the lambs in the control group. These results indicate that lactogenic immunoprophylaxis should help mitigate the financial losses caused by cryptosporidiosis in small ruminants, as well as reducing the risk of infection of humans through the decreased contamination of the environment with oocysts. PMID- 15817198 TI - Serological responses to Neospora caninum in experimentally and naturally infected water buffaloes (Bubalus bubalis). AB - The water buffalo (Bubalus bubalis) is an important natural host for Neospora caninum. Serologic responses to N. caninum were studied in experimentally and naturally infected water buffaloes in Brazil. Antibodies were assayed by the indirect fluorescent antibody test (IFAT) using a cut off value of 1:25. Six buffaloes were each inoculated subcutaneously with 5 x 10(6) live culture-derived tachyzoites of the cattle Illinois strain of N. caninum, and two calves were kept as uninoculated controls. Post-inoculation (p.i.) blood samples were collected weekly for 8 weeks and then monthly until 1 year p.i. All inoculated buffaloes developed IFAT titers of 1:100 or more between 7 and 11 days p.i. and the titers remained elevated until 7 weeks p.i. Antibody titers peaked to 1:1600 in 1, 1:800 in 3 and 1:400 in 2, usually by 3 weeks p.i. Antibody titers declined to 1:25 or 1:50 in all the six buffaloes by 12 months p.i. IFAT titers to N. caninum remained at an undetectable level (< 1:25) in both control uninoculated buffaloes. To follow the dynamics of N. caninum antibodies, sera from 29 buffaloes and their calves were collected for 1 year and assayed for N. caninum antibodies; 23 of 29 calves were seropositive (IFAT of 1:100 or more) at 1-2 day of age. Of these 23 calves, 17 remained seropositive during the study, while six became seronegative at four (two calves), six (one calf) seven (two calves) and eight (one calf) months of age. These findings suggest a high rate of neonatal transmission of N. caninum in buffaloes. PMID- 15817199 TI - Immunisation of mice against neosporosis with recombinant NcSRS2 iscoms. AB - The coccidian parasite Neospora caninum is an intracellular protozoan, causing abortion in cattle in many countries around the world. In this study, the protective potential of the major N. caninum surface antigen NcSRS2, expressed in Escherichia coli and formulated into immunostimulating complexes (iscoms), was investigated in an experimental mouse model. The recombinant protein was specially designed for binding to iscoms via biotin-streptavidin interaction. Two groups of 10 BALB/c mice were immunised twice, on days 0 and 28 with iscoms containing either the recombinant NcSRS2 (NcSRS2 iscoms) or similar iscoms with NcSRS2 substituted by an unrelated recombinant malaria peptide (M5) as a control (M5 iscoms). A third group of 10 age-matched BALB/c mice served as an uninfected control group. Immunisation with recombinant NcSRS2 iscoms resulted in production of substantial antibody titres against N. caninum antigen, while the mice immunised with M5 iscoms produced only very low levels of antibodies reacting with N. caninum antigen. After challenge infection with N. caninum tachyzoites on day 69, mice immunised with NcSRS2 iscoms showed only mild and transient symptoms, whereas the group immunised with M5 iscoms showed clinical symptoms until the end of the experiment at 31 days post inoculation. A competitive PCR assay detecting Nc5-repeats was applied to evaluate the level of parasite DNA in the brain. The amount of Nc5-repeats in the group vaccinated with NcSRS2 iscoms was significantly lower than in the control group given M5 iscoms. In conclusion, it was found that the recombinant NcSRS2 iscoms induced specific antibodies to native NcSRS2 and immunity sufficient to reduce the proliferation of N. caninum in the brains of immunised mice. PMID- 15817200 TI - Individual intake and antiparasitic efficacy of free choice mineral containing fenbendazole for grazing steers. AB - A 95-day study (June 25-September 27, 2001) was conducted using 120 steers (311.9+/-2.4 kg) randomly allocated to two treatments: (1) mineral containing 0.55% fenbendazole (FBZ) and (2) control, no FBZ in the mineral. Animals in the FBZ group were individually identified by an electronic tag that was read each time an animal attended the mineral feeder. The feeder was equipped with load cells that enabled individual mineral intakes to be estimated. The FBZ group was provided with non-medicated mineral during a 14-day adaptation period (July 23 August 5) and an 8-day post-medication period (September 17-24). The intake of FBZ was monitored for 14 days during each of the two treatment periods; August 6 19 and September 3-16, separated by a 14-day non-medicated period, August 20 September 2. Control animals had access to non-medicated mineral for the entire 95-day study period. All steers were grazed on alfalfa-grass pasture for the duration of the study and had free access to flocculated, filtered and chlorinated water via an automatic waterer. Fecal samples were collected from steers three times during the experiment July 23, August 27 and September 27, and analyzed for nematode eggs and Giardia sp. cysts. Seventy-five and 83% of the steers in the FBZ group visited the mineral feeder during the first and second treatment periods, respectively. Individual daily mineral and FBZ intake for the first and second treatment periods was 52.9+/-6.6g per day and 10.1+/-1.2mg/kg BW; 72.3+/-8.4 g per day and 11.8+/-1.4 mg/kg BW, respectively. FBZ animals were separated into three groups during each treatment period based on the recommended dose (RD) of FBZ (5 mg/kg/BW), those that received > the RD, those that received < RD but > 50% RD and those that received < 50% of RD. Nematode egg counts and the number of animals infected with nematodes was reduced (p < 0.05) in all cattle that consumed FBZ as compared to control animals. In contrast to nematode eggs, numbers of Giardia cysts was not reduced (p > 0.05) by FBZ as compared to controls in either treatment period. These results may be a reflection of Giardia re-infection occurring following treatment and highlight the need for variation in treatment regimes specifically targeted at the parasite of interest. PMID- 15817201 TI - Development of a single-round and multiplex PCR method for the simultaneous detection of Babesia caballi and Babesia equi in horse blood. AB - With the aim of developing more simple diagnostic alternatives, a differential single-round and multiplex polymerase chain reaction (PCR) method was designed for the simultaneous detection of Babesia caballi and Babesia equi, by targeting 18S ribosomal RNA genes. The multiplex PCR amplified DNA fragments of 540 and 392 bp from B. caballi and B. equi, respectively, in one reaction. The PCR method evaluated on 39 blood samples collected from domestic horses in Mongolia yielded similar results to those obtained from confirmative PCR methods that had been established earlier. Thus, the single-round and multiplex PCR method offers a simple tool for the differential diagnosis of B. caballi and B. equi infections in routine diagnostic laboratory settings as well as in epidemiological studies. PMID- 15817202 TI - Osteolytic osteomyelitis associated with visceral leishmaniasis in a dog. AB - A dog was examined with a history of weight loss and lameness of the left hind limb. A painful response to examination of the left hip joint, and lymphadenopathy were noted. Amastigote forms of Leishmania sp. were observed by cytology in samples from the popliteus lymph node, and anti-Leishmania sp. antibodies at a titer of 1:640 were detected in serum by indirect immunofluorescence. Radiological changes included osteolysis and a periosteal proliferative reaction in the left femoral greater trochanter. These changes were histologically characterized as an osteolytic granulomatous osteomyelitis associated with amastigotes within macrophages. Non-decalcified fragments of the periosteum were processed for immunohistochemistry, observed with prominent immunolabelling of amastigotes of Leishmania sp. within macrophages. The diagnosis was further confirmed by positive PCR for Leishmania sp., belonging to the Leishmania donovani complex. PMID- 15817203 TI - The influence of colostrum on infection of calves around 7 months of age with Schistosoma mattheei. AB - Studies have indicated that the intake of colostrum could modulate the offspring reaction towards early schistosome infections. The effect of colostrum (containing immunoglobulins, parasite antigens, immune cells and other cell related products) on late Schistosoma infections is to our knowledge not documented. The objective of the present study is to determine whether the intake of colostrum from Schistosoma mattheei infected cows will modify late S. mattheei infection patterns in their offspring. Six calves born to confirmed non-infected cows and 10 calves born to confirmed infected mothers were purchased after intake of colostrum. All calves were exposed to a total experimental challenge of 2500 cercariae around the age of 7 months. Serum samples were collected before and after intake of colostrum and monthly thereafter for the determination of specific antibody levels. Faecal samples were collected monthly from 42 days after infection for the determination of faecal egg counts. Six calves of each group were slaughtered around the age of 15 months for worm recovery and tissue egg counting. No differences between both groups were observed in immunoglobulin levels and faecal egg counts after infection, and in worm counts and tissue egg counts at necropsy. In conclusion colostral effects, which were noticed at an early age, are no longer present around the age of 7 months. As such calves which are born during a season of high Schistosoma transmission will still be under colostral influence and therefore be more protected against a primary challenge than calves born during a low transmission season, as the latter will only receive their first challenge when colostral protective effects have disappeared. PMID- 15817204 TI - Performance characteristics of an enzyme-linked immunosorbent assay performed in milk for the detection of liver fluke (Fasciola hepatica) infection in cattle. AB - AIM: To determine the performance characteristics of an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against liver fluke (Fasciola hepatica) in bovine milk. METHODS: Serum and milk from liver fluke infected and non-infected cattle was assayed in a commercially available enzyme linked immunosorbent assay. Serum test results were used to determine the "gold standard" infection status of cattle and milk ELISA results assessed by ROC analysis. RESULTS: ROC analysis suggested changes to the ELISA protocol, arriving at milk dilutions assayed considerably higher than those suggested by the manufacturer. With those changes, the ELISA performed with high sensitivity and specificity, 95 and 98.2%, respectively, for individual bovine milks (relative to sera). For bovine tank milks, sensitivity was lower, with bulk milks only testing positive if 60% or more of cattle milking in the herd were infected. CONCLUSIONS: The analysis of the ELISA's performance when used on individual bovine milks demonstrated high sensitivity and specificity. ROC analyses optimised the assay conditions and cut-off point suggested by the manufacturer for this commercial diagnostic assay. This would help with the identification and control of fasciolosis, enabling simpler sample collection. PMID- 15817205 TI - A survey to determine relationships between bulk tank milk antibodies against Ostertagia ostertagi and milk production parameters. AB - In temperate climate regions, gastrointestinal nematodes are still widespread in adult dairy cows, but until now there exists no reliable diagnostic tool that can identify herds where the infection interferes with productivity. The objective of this study was to investigate the relationships between levels of antibodies against Ostertagia ostertagi in bulk tank milk and milk production. Bulk tank milk samples of 2553 dairy herds were obtained in spring and 2104 of these herds were sampled a second time in autumn. The antibody levels against O. ostertagi were determined with a milk ELISA and test results were expressed as an optical density ratio (ODR). The effect of bulk tank milk ODR on three different production parameters, kg milk, % and kg fat, % and kg protein was assessed by a multivariable linear regression model on the herds for which production data were available (n = 1063 and 867 in spring and autumn, respectively). The mean and standard deviation for ODRautumn (0.972+/-0.238) were higher than for ODRspring (0.825+/-0.201). Significant negative relationships were found between ODR and milk yield. An increase in ODRspring and ODRautumn from the 25th to the 75th percentile of the available ODR data was associated with a drop in the annual milk yield of 1.1 kg/cow/day, respectively 0.9 kg/cow/day. When a herd's ODR increased between spring and autumn with 0.142, it produced on average 0.4 kg/cow/day less in September than in April, in comparison with herds where the ODR did not increase. A significant negative association was found between ODRautumn and % protein averaged over the period of a year. No significant associations were found between ODR and % fat averaged over the year. When protein and fat production of September were expressed in kg an increase in ODRautumn from the 25th to the 75th percentile was associated with a decrease of 0.037 kg protein/cow/day and 0.042 kg fat/cow/day. PMID- 15817206 TI - Exsheathment of Ostertagia ostertagi infective larvae following exposure to bovine rumen contents derived from low and high roughage diets. AB - The objective of this study was to characterize the exsheathment kinetics of Ostertagia ostertagi infective larvae (L3) following in vivo exposure to bovine rumen contents derived from low and high roughage diets. O. ostertagi L3 were placed in disposable dialysis bags and incubated for various time points between 0 and 360 min in the rumen of a fistulated steer maintained on a 71% grain diet or a 100% grass diet. The maximum percentage of exsheathed L3 was observed 120 min post-exposure to grass-derived rumen contents, while maximum exsheathment for L3 exposed to grain-derived rumen contents did not occur until 360 min. This work provides the first report of the in vivo exsheathment kinetics for O. ostertagi in its bovine host. Results of this study also support earlier reports that rumen pH may affect the exsheathment efficiency of abomasal trichostrongylids. PMID- 15817207 TI - The effect of free and polyethylene glycol-liposome-entrapped albendazole on larval mobility and number in Toxocara canis infected mice. AB - As part of our exploratory drug research on the larva migrans that causes roundworm in dogs and cats, this study was carried out to clarify the effect of free and liposome-entrapped (LE) albendazole in Toxocara canis infected mice. In infected mice, evaluation of mobility and number of larva were examined in detail in the brain, skeletal muscle and liver. Larva mobility was evaluated by using the relative mobility (RM) value. Albendazole was LE as one of the drug delivery systems (DDSs). Polyethylene glycol (PEG) was added to the liposome in order to avoid evoking a response by the reticuloendothelial system (RES). By using the albendazole PEG-LE delivery system, it was possible to target the larvae in the mouse brain and liver resulting in a decrease in the number of larvae. In the skeletal muscle of the infected mice, the intraperitoneal dosages of PEG-LE albendazole did not cause a complete decrease in the number of larvae, even though free albendazole did cause the number to decrease. Therefore, it is necessary to take into consideration the migrating stage of the larvae before the initiation of any drug administration. PMID- 15817208 TI - Treatment of dog thelaziosis caused by Thelazia callipaeda (Spirurida, Thelaziidae) using a topical formulation of imidacloprid 10% and moxidectin 2.5%. AB - Canine thelaziosis caused by Thelazia callipaeda infects dogs, cats, foxes, rabbits, and humans resulting in conjunctivitis, pain and excessive lacrimation. T. callipaeda live in the eyes under the nictitating membrane and females release first stage larvae which are ingested by flies which act as intermediate hosts. Control of canine thelaziosis is currently based on the removal of nematodes directly from the eyes of affected dogs or on the local instillation of antiparasitic drugs. With the aim of evaluating the efficacy against T. callipaeda of an association of imidacloprid 10% and moxidectin 2.5% by spot-on formulation administered via dermal application, three groups of naturally infected animals were selected: i.e. group A (21 dogs) received a single dose of imidacloprid 10% and moxidectin 2.5% by spot-on; group B (21 dogs) received a single dose of imidacloprid 10% by spot-on and group C (20 dogs) were left untreated. The efficacy of treatments was established by eye inspection and parasite viability and vitality after 1, 5 and 9 days after animal treatments (groups A and B) and after 9 days only in untreated animals (group C). Imidacloprid 10% and moxidectin 2.5% in spot-on formulation showed to be effective with regards the control of dog thelaziosis within 5 (90.47%) to 9 (95.23%) days after treatment. Only one dog from group A presented nematodes after treatment. The presence of parasites in the eyes of dogs from groups B (imidacloprid 10%) and C confirm that the anthelmintic efficacy against T. callipaeda in animal from group A (imidacloprid 10% and moxidectin 2.5%) was most likely attributable to moxidectin 2.5%. The spot-on formulation containing imidacloprid 10% and moxidectin 2.5% is very easy to apply and helps overcome problems linked to the mechanical removal of parasites or to the restraining of the animals for the local instillation of drugs in the eyes. PMID- 15817209 TI - The possibilities and limitations of evasive grazing as a control measure for parasitic gastroenteritis in small ruminants in temperate climates. AB - Between 1999 and 2003 studies were done at Utrecht University on the population dynamics of gastrointestinal nematodes and the potential to use evasive grazing for the control of nematode infections in small ruminants. On most pastures grazed in May and June it took at least 3 weeks before high pasture infectivity levels for Haemonchus contortus developed while it took at least 2 weeks on pastures grazed in July, August and September. Occasionally, larvae emerged earlier on pasture when high temperatures coincided with rain. Pasture infectivity levels for H. contortus subsequently increased and the highest levels were found between 5 and 9 weeks on pastures grazed in May-June and between 3 and 9 weeks on pastures grazed in July, August and September. Pasture infectivity usually had decreased again to low levels after approximately 3 months. Larvae of the other trichostrongylids emerged later on pasture. However, patterns of Teladorsagia circumcincta, Trichostrongylus spp. and Cooperia curticei were basically similar to H. contortus. Strongyloides papillosus larvae emerge within 2 weeks on pasture and survival is short. On the other hand Nematodirus larvae took a lot longer to emerge on pasture than the other trichostrongylids. The results imply that only on a small proportion of farms (mixed dairy cattle/sheep farms; some organic dairy goat farms) evasive grazing can be recommended as only control measure for parasitic gastroenteritis. However, the present data offer possibilities to combine evasive grazing with other methods in an integrated control scheme. PMID- 15817210 TI - Effects of protein supply and reproductive status on local and systemic immune responses to Teladorsagia circumcincta in sheep. AB - The effects of protein supply and reproductive status on circulating antibody responses and local inflammatory cell counts were investigated in parasitized sheep, with local immune responses assessed through a recently refined abomasal cannulation methodology. We hypothesized that if breakdown of immunity has a nutritional basis, then protein scarcity would result in a breakdown of immunity to Teladorsagia circumcincta in both periparturient and non-reproducing (barren) ewes. Twin-bearing and barren, abomasally cannulated ewes were fed at either 0.8 or 1.3 times protein requirements from 3 weeks before until 6 weeks after parturition (n = 6). All sheep were trickle infected at a rate of 10,000 infective larvae (L3) per day, for 3 days per week throughout the experiment. Faecal egg counts remained virtually zero in all barren ewes, whilst protein supplementation reduced faecal egg counts in the periparturient ewes during most of the periparturient period. Final worm burdens, taken at 6 weeks into lactation, were lower for the barren ewes than for the lactating ewes, whilst protein supplementation reduced worm burdens in the latter. Protein supply did not affect mucosal mast cell counts, which were consistently higher for the barren ewes than the periparturient ewes, but were temporarily decreased around parturition. Barren ewes and protein supplemented lactating ewes had higher globule leukocyte counts than the unsupplemented lactating ewes. Protein supplementation increased eosinophil counts in the lactating ewes though only during the later part of the lactation period. Plasma IgA anti-L3 antibody was similar for all ewes, but IgE anti-L3 antibody was higher for the protein supplemented periparturient ewes compared to the unsupplemented periparturient ewes and all barren ewes. It is likely that the combination of low protein requirements and large body protein reserves did not result in breakdown of immunity to T. circumcincta for the barren ewes. These results suggest that changes in mucosal mast cell and eosinophil counts are not necessarily associated with changes in host resistance to T. circumcincta. However, the data support the view that increased globule leukocyte counts and plasma IgE anti-L3 antibody may be associated with nutritionally improved expression of immunity in periparturient ewes. PMID- 15817212 TI - Ixodid tick species attaching to dogs in Hungary. AB - A survey was carried out to investigate the occurrence of hard tick species (Acari: Ixodidae) infesting domestic dogs in Hungary. Forty veterinary clinics from a wide geographical area were asked to collect hard ticks from dogs and to complete a questionnaire. In total, 25 veterinary clinics submitted 900 ticks from 310 dogs. Intensity of infestation ranged from one to 78 per dog. The most preferred sites of tick attachment in decreasing order were head, neck and legs. The majority of ticks (91.7%) were adults, which were identified to species level, the others were nymphs. Six species were found: Dermacentor reticulatus (48.9%), Ixodes ricinus (43.2%), Ixodes canisuga (5.6%), Haemaphysalis concinna (2%) and there was one specimen of both Dermacentor marginatus and Ixodes hexagonus. Single species infestation with I. ricinus or D. reticulatus was found on 145 (46.8%) and 120 animals (38.7%), respectively. Mixed infestation caused by these two species was detected on 24 dogs (7.7%). I. canisuga and H. concinna were found on seven and five dogs, respectively. D. reticulatus and I. ricinus were collected almost throughout the year, except for a single month. The activity peaks were in spring and in autumn for both species. Based on clinical signs, canine babesiosis was diagnosed by the veterinarians in 66 (21.3%) tick infested dogs. These dogs were more frequently infested with D. reticulatus than the others. Our data contribute to the understanding of geographical and seasonal distribution of ixodid tick species infesting dogs in Hungary. The implication of these data, for the risk of canine tick borne diseases is discussed. PMID- 15817211 TI - Detection of antibody to Toxocara vitulorum perieneteric fluid antigens (Pe) in the colostrum and serum of buffalo calves and cows by Western blotting. AB - Toxocara vitulorum, a nematode parasite in the small intestine of cattle and water buffaloes, causes high morbidity and mortality of 1-3 months old buffalo calves. This research evaluated the specific perieneteric antigens (Pe) reactivity of anti-T. vitulorum-Pe antibody (Tv-Pe-Ab) in both immune sera and colostrum from buffalo cows immediately post-partum from buffalo cows. The presence of Tv-Pe-Ab in sera of buffalo newborn calves was also examined at 1 day before and after suckling the colostrum as well as in sera from naturally infected calves at the beginning and peak of the maximum infection and then again during the period of rejection and post-rejection of the parasite. Pe antigens were characterized for Tv-Pe-Ab by SDS-PAGE and Western blot (WB). The SDS-PAGE showed that Pe contained nine protein bands (11, 14, 31, 38, 58, 76, 88, 112 and 165 kDa). All Pe bands were recognized by Tv-Pe-Ab in sera and colostrum of buffalo cows. Only the serum antibodies of buffalo calves at 1 day of age after suckling the colostrum and during the beginning of T. vitulorum infection recognized Pe antigen's nine bands. In contrast, serum antibodies from 1-day-old buffalo calves, taken before suckling colostrum, did not react with any protein band. In suckling calves, which reached peak egg output, rejection and post rejection stages of the infection, serum Tv-Pe-Ab reactivity with lower molecular weight protein bands (11-76 kDa) was lost and only reactivity with the Pe protein bands of higher molecular weight (88, 112 and 165 kDa) remained. PMID- 15817213 TI - Sero-epidemiological surveillance of hypodermosis in yaks and cattle in north China by ELISA. AB - A large-scale serological surveillance was conducted for hypodermosis in the north of China. A total of 4175 sera samples of yaks and cattle were collected from five provinces in north China that included Xinjiang, Inner Mongolia, Heilongjiang, Jiling and Gansu provinces, between 2001 and 2002, and were examined for anti-hypoderma antibody by ELISA. The results indicate that the naturally infested positive rates of Hypoderma spp. in the above provinces were 51.77%, 27.02%, 13.00%, 6.03% and 44.41%, respectively. These results suggest a significant warble fly population in the north of China. Positive sera were collected from infested cattle in Pingliang county of the Gansu province monthly between August 2001 and July 2002. These sera were used to evaluate the seasonal kinetics of anti-hypoderma antibody. The kinetics demonstrated that anti hypoderma antibody was elevated from October to December. Thus, serological surveillances associated with low infestation rates and chemical therapy would best be performed from October to December in the region. PMID- 15817214 TI - Comparison of the external morphology of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks from Brazil and Argentina. AB - In the present study the external morphology of semi-engorged Rhipicephalus sanguineus females ticks from Brazil and Argentina were compared by scanning electron microscopy. Many differences were detected among the R. sanguineus collected at the two localities, such as body size, shape of the genital pore, and morphology of the sensory structures. All these characteristics are fundamental for the diagnosis of species of the genus Rhipicephalus and thus indicate the need for further comparisons and, the taxonomical revision of this species of tick in the Neotropics. PMID- 15817215 TI - Association of the kdr and superkdr sodium channel mutations with resistance to pyrethroids in Louisiana populations of the horn fly, Haematobia irritans irritans (L.). AB - Pyrethroid resistance in three horn fly populations in Louisiana was monitored by weekly fly counts, filter paper bioassays, and diagnostic PCR assays for the presence of pyrethroid resistance-associated mutations in the sodium channel gene coding region. The PCR assay for the knockdown resistance (kdr) and superkdr sodium channel mutations was used to determine the frequency of the target site insensitivity mechanism in the populations of horn flies, which possessed varying degrees of insecticide resistance. The bioassays and frequency of homozygous resistant (RR) kdr genotypes were relative predictors of the fly control subsequently observed. Flies exposed to filter paper impregnated with a discriminating concentration of one of four different insecticides were collected when 50% mortality was estimated. Genotypes for the dead flies and the survivors were determined by the PCR assay. The results of the PCR assays indicated that the genotype at the kdr locus of the flies exposed to the two pyrethroids had an effect upon whether the flies were considered to be alive or dead at the time of collection. The kdr genotype of flies exposed to either diazinon or doramectin was unrelated to whether the flies were considered to be alive or dead, except for a single comparison of flies exposed to diazinon. When possible interactions of the kdr and superkdr mutations were compared, we found that there were no associations with the response to diazinon and doramectin. For one location, there were no survivors of the 75 flies with the SS-SS (superkdr-kdr) homozygous susceptible wild type genotype exposed to pyrethroids, while there were survivors in all of the other five genotypes. The SS-RR genotype flies were more susceptible to the pyrethroids than the SR-RR flies, but that was not the case for exposure to diazinon or doramectin. In the St. Joseph population, there were an adequate number of flies to demonstrate that the SS-SR genotype was more susceptible to pyrethroids than the SS-RR and that flies with the SR-SR genotype were more susceptible to pyrethroids than the flies with the SR-RR genotype. PMID- 15817216 TI - Relationship between type 1/type 2 immune responses and occurrence of vertical transmission in BALB/c mice infected with Neospora caninum. AB - To examine the relationship between occurrence of vertical transmission and type 1/type 2 immune responses induced by Neospora caninum infection in BALB/c mice, pregnant (group 1 p) and non-pregnant mice (group 1 np) were inoculated with 2 x 10(6) of the N. caninum parasites and then we examined the vertical transmission rate and production of IFN-gamma and IL-4. We also studied chronically infected mice, which were bred at 4 weeks or more after infection (group 2), and mice inoculated during pregnancy and re-bred at 4 weeks or more after delivery (group 3). In groups 1p, 2 and 3, vertical transmission was observed in 27.4, 41.4, and 50% of the offspring, respectively. The serum IFN-gamma level increased on days 1 and 5 post-inoculation (p.i.) in groups 1 p and 1 np, while no increase level was observed in groups 2 and 3 during pregnancy or after delivery. When the mice in groups 2 and 3 were re-inoculated, all mice showed a transient increase in serum IFN-gamma on day 1 post-re-inoculation. The serum IL-4 level in both of groups 1p and np increased in a similar manner following infection. In group 3, the serum IL-4 level was somewhat higher than that in group 2 after re-inoculation. The anti-N. caninum antibody IgG1 titer in group 3 increased on day 10 post-re inoculation. These results suggest that the mice infected during pregnancy may acquire a weaker immune response to the parasite than mice infected when they are not pregnant, and that mice infected during pregnancy may show an enhanced type 2 immune response in the recrudescence of the infection. PMID- 15817217 TI - Infectivity of a novel type of Cryptosporidium andersoni to laboratory mice. AB - Previously, we reported 'a novel type' of Cryptosporidium andersoni detected from cattle in Japan, and showed that the isolate was infective to mice. In the present study, we examined the patterns of oocyst shedding in both immunocompromised and immunocompetent mice, as well as pathological lesions in the infected mice. After oral inoculation with 1 x 10(6) oocysts, all five severe combined immunodeficiency (SCID) mice began to shed endogenously produced oocysts on day 6 post-inoculation (p.i.). The number of oocysts per day (OPD) reached 1 x 10(6) on day 17 p.i., and an OPD level of 1 x 10(6) to 10(7) was maintained until 91 days p.i. when the mice were sacrificed. In the five immunocompetent mice inoculated with 1 x 10(6) oocysts, the pre-patent and patent periods were 6 and 19 days, respectively, and the maximal OPD level was 1.5 x 10(5) on average. On histological examinations of infected SCID mice, a large number of parasites were present on the surface of the gastric glands of the stomach, but not in other organs examined. In conclusion, the novel type of C. andersoni, which genetically coincides with C. andersoni reported in other countries, is infective to mice, but susceptibility was lower than that of Cryptosporidium muris infecting rodents from the perspective of infectivity to immunocompetent mice. PMID- 15817218 TI - Observations on helminth infections of free-living and captive rheas (Rhea americana) in Brazil. AB - The present work describes helminth infection of eight free-living and 12 captive rheas (Rhea americana) from, respectively, Pantanal of Mato Grosso do Sul State, and Jaboticabal, Sao Paulo State, Brazil. Captive birds were young and had a high mortality rate, while free-living birds were adult and apparently healthy. Infections were evaluated by post-mortem examination of internal organs and recovery of helminths using standard parasitological procedures. Seven species of nematodes (Sicarius uncinipenis, Torquatoides crotophaga, Deletrocephalus dimidiatus, D. cesarpintoi, Paradeletrocephalus minor, Capillaria venteli and Dicheilonema rheae) and two species of cestodes (Houttuynia struthionis and Chapmania tauricolis) were identified. P. minor, which inhabits the large intestine, was the most common helminth in free-living birds (63.9%). In captive rheas, a mean parasitic load of 173 helminths per host was found. The gizzard of these birds was the most parasitized organ and S. uncinipenis was most common (92.5%). Parasitism of free-living and captive birds and associated pathology are discussed. PMID- 15817219 TI - Acaricidal properties of the essential oil of Melaleuca alternifolia Cheel (tea tree oil) against nymphs of Ixodes ricinus. AB - The aim of the study was to examine the acaricidal effect of essential oil of Melaleuca alternifolia (tea tree oil, TTO) at different doses (4, 6, 8 and 10 microl) and for different exposure times (30, 60, 90 and 120 min) on nymphs of Ixodes ricinus. A dose of 8 microl TTO was lethal for more than 70% of ticks when inhaled and this effect was enhanced when the dose was increased to 10 microl (> 80%). The effect was correlated with the duration of exposure of ticks to TTO, with a significant effect being observed after 90 min exposure. The findings show that TTO has acaricidal properties and could be extremely useful in controlling ticks that are efficient vectors of pathogens. PMID- 15817220 TI - Eya1 acts upstream of Tbx1, Neurogenin 1, NeuroD and the neurotrophins BDNF and NT-3 during inner ear development. AB - Cell fate specification during inner ear development is dependent upon regional gene expression within the otic vesicle. One of the earliest cell fate determination steps in this system is the specification of neural precursors, and regulators of this process include the Atonal-related basic helix-loop-helix genes, Ngn1 and NeuroD and the T-box gene, Tbx1. In this study we demonstrate that Eya1 signaling is critical to the normal expression patterns of Tbx1, Ngn1, and NeuroD in the developing mouse otocyst. We discuss a potential mechanism for the absence of neural precursors in the Eya1-/- inner ears and the primary and secondary mechanisms for the loss of cochleovestibular ganglion cells in the Eya1bor/bor hypomorphic mutant. PMID- 15817221 TI - XNGNR1-dependent neurogenesis mediates early neural cell death. AB - Early neural cell death is programmed cell death occurring within proliferating and undifferentiated neural progenitors. Little is known about the regulation and role of early neural cell death. In Xenopus embryos, primary neurogenesis is disrupted following the inhibition of early neural cell death, indicating that it is required for normal primary neurogenesis. Here we show that early neural cell death is dependent on primary neurogenesis. Overexpression of XSoxD concomitantly reduced N-Tubulin expression and early neural cell death, as seen by reduced TUNEL staining in stage 15 embryos. Conversely, overexpression of XNgnr1 led to ectopic N-Tubulin expression and TUNEL staining. However, XNeuroD overexpression, which induces ectopic N-Tubulin expression downstream of XNgnr1, had no effect on early neural cell death. E1A12S differentially inhibits the differentiation pathway induced by XNGNR1 protein. E1A12S-mediated inhibition of XNGNR1 neurogenic activity resulted in the reduction of N-Tubulin expression and TUNEL staining. Taken together, our data establish that primary neurogenesis induced by XNGNR1 promotes early neural cell death. This indicates that XNgnr1 positively regulates early neural cell death. We propose that early neural cell death might eliminate cells with abnormally high levels of XNGNR1, which can result in pre mature neuronal differentiation. PMID- 15817222 TI - Nanos downregulates transcription and modulates CTD phosphorylation in the soma of early Drosophila embryos. AB - nanos (nos) specifies posterior development in the Drosophila embryo by repressing the translation of maternal hb mRNA. In addition to this somatic function, nos is required in the germline progenitors, the pole cells, to establish transcriptional quiescence. We have previously reported that nos is required to keep the Sex-lethal establishment promoter, Sxl-Pe, off in the germline of both sexes. We show here that nos also functions to repress Sxl-Pe activity in the surrounding soma. Sxl-Pe is inappropriately activated in the soma of male embryos from nos mothers, while Sxl-Pe can be repressed in female embryos by ectopic Nos protein. nos appears to play a global role in repressing transcription in the soma as the effects of nos on promoter activity are correlated with changes in the phosphorylation status of the carboxy terminal domain (CTD) repeats of the large RNA polymerase II subunit. Finally, we present evidence indicating that the suppression of transcription in the soma by Nos protein is important for normal embryonic development. PMID- 15817223 TI - Phox2b function in the enteric nervous system is conserved in zebrafish and is sox10-dependent. AB - Zebrafish lacking functional sox10 have defects in non-ectomesenchymal neural crest derivatives including the enteric nervous system (ENS) and as such provide an animal model for human Waardenburg Syndrome IV. Here, we characterize zebrafish phox2b as a functionally conserved marker of the developing ENS. We show that morpholino-mediated knockdown of Phox2b generates fish modeling Hirschsprung disease. Using markers, including phox2b, we investigate the ontogeny of the sox10 ENS phenotype. As previously shown for melanophore development, ENS progenitor fate specification fails in these mutant fish. However, in addition, we trace back the sox10 mutant ENS defect to an even earlier time point, finding that most neural crest cells fail to migrate ventrally to the gut primordium. PMID- 15817224 TI - Notch signaling modulates the nuclear localization of carboxy-terminal phosphorylated smad2 and controls the competence of ectodermal cells for activin A. AB - Loss of mesodermal competence (LMC) during Xenopus development is a well known but little understood phenomenon that prospective ectodermal cells (animal caps) lose their competence for inductive signals, such as activin A, to induce mesodermal genes and tissues after the start of gastrulation. Notch signaling can delay the onset of LMC for activin A in animal caps [Coffman, C.R., Skoglund, P., Harris, W.A., Kintner, C.R., 1993. Expression of an extracellular deletion of Xotch diverts cell fate in Xenopus embryos. Cell 73, 659-671], although the mechanism by which this modulation occurs remains unknown. Here, we show that Notch signaling also delays the onset of LMC in whole embryos, as it did in animal caps. To better understand this effect and the mechanism of LMC itself, we investigated at which step of activin signal transduction pathway the Notch signaling act to affect the timing of the LMC. In our system, ALK4 (activin type I receptor) maintained the ability to phosphorylate the C-terminal region of smad2 upon activin A stimulus after the onset of LMC in both control- and Notch activated animal caps. However, C-terminal-phosphorylated smad2 could bind to smad4 and accumulate in the nucleus only in Notch-activated animal caps. We conclude that LMC was induced because C-terminal-phosphorylated smad2 lost its ability to bind to smad4, and consequently could not accumulate in the nucleus. Notch signal activation restored the ability of C-terminal-phosphorylated smad2 to bind to smad4, resulting in a delay in the onset of LMC. PMID- 15817225 TI - Co-operation between enhancers modulates quantitative expression from the Drosophila Paramyosin/miniparamyosin gene in different muscle types. AB - The distinct muscles of an organism accumulate different quantities of structural proteins, but always maintaining their stoichiometry. However, the mechanisms that control the levels of these proteins and that co-ordinate muscle gene expression remain to be defined. The paramyosin/miniparamyosin gene encodes two thick filament proteins transcribed from two different promoters. We have analysed the regulatory regions that control expression of this gene and that are situated in the two promoters, the 5' and the internal promoters, both in vivo and in silico. A distal muscle enhancer containing three conserved MEF2 motifs is essential to drive high levels of paramyosin expression in all the major embryonic, larval and adult muscles. This enhancer shares sequence motifs, as well as its structure and organisation, with at least four co-regulated muscle enhancers that direct similar patterns of expression. However, other elements located downstream of the enhancer are also required for correct gene expression. Other muscle genes with different patterns of expression, such as miniparamyosin, are regulated by other basic mechanisms. The expression of miniparamyosin is controlled by two enhancers, AB and TX, but a BF modulator is required to ensure the correct levels of expression in each particular muscle. We propose a mechanism of transcriptional regulation in which similar enhancers are responsible for the spatio-temporal expression of co-regulated genes. However, it is the interaction between enhancers which ensures that the correct amounts of protein are expressed at any particular time in a cell, adapting these levels to their specific needs. These mechanisms may not be exclusive to neural or muscle tissue and might represent a general mechanism for genes that are spatially and temporally co-regulated. PMID- 15817226 TI - Structural and functional evidences for a type 1 TGF-beta sensu stricto receptor in the lophotrochozoan Crassostrea gigas suggest conserved molecular mechanisms controlling mesodermal patterning across bilateria. AB - The transforming growth factor beta (TGFbeta) superfamily includes bone morphogenetic proteins, activins and TGF-betasensu stricto (s.s.). These ligands have been shown to play a key role in numerous biological processes including early embryonic development and immune regulation. They transduce their signal through a hetromeric complex of type I and type II receptors. Such receptors have been identified in ecdysozoans but none have been found as yet in the other major protostomal clade, the lophotrochozoans. Here, we report the identification of the first lophotrochozoan TGFbetas.s. type I receptor (Cg-TGFbetaRI) from the mollusk Crassostrea gigas. The phylogenetic and structural analyses as well as the expression pattern during early development suggest Cg-TGFbetaRI to belong to the TGFbetas.s./activin type I receptor clade and functional studies corroborate these deductions. The use of the zebrafish embryo as a reporter organism reveals that either Cg-TGFbetaRI or its dominant negative acting truncated form, when overexpressed during gastrulation, resulted in a range of phenotypes displaying severe disturbance of anterioposterior patterning due to a strong modulation of ventrolateral mesoderm patterning. Finally, a Cg-TGFbetaRI cytokine activity during immune regulation in C. gigas has been investigated by real-time PCR in haemocytes and mantle edge during an in vivo bacterial LPS challenge. One piece of evidence from this study suggests that the molecular mechanisms controlling mesodermal patterning and some immune regulations across all bilateria could be conserved through a functional TGF-beta s.s. pathway in lophotrochozoans. PMID- 15817227 TI - RNA helicase A is important for germline transcriptional control, proliferation, and meiosis in C. elegans. AB - RNA helicase A (RHA) is a multifunctional protein with established roles in chromatin regulation. The protein is conserved in worms, Drosophila, and mammals, but its role in worms has not been previously studied. We found that a deletion mutant lacking rha-1 has a temperature-sensitive defect in germline transcriptional silencing, consistent with RHA-1 having a function in transcription regulation. Transcriptional desilencing in these rha-1(tm329) mutants was associated with a loss of lysine 9 methylation on histone H3 that is normally associated with silenced chromatin. Other histone modifications are also mis-localized in the germ cells in the mutants. These defects in histone modifications suggest that there is a general transcription regulation defect in the mutant worms that results in a temperature-sensitive sterile phenotype. At the restrictive temperature, the extent of germ cell mitoses is reduced, and the mutants are sterile due to defects in meiosis and gametogenesis. Our results suggest that RHA-1 is a conserved transcription regulation protein that controls germline proliferation and development in C. elegans. PMID- 15817228 TI - Regeneration and asexual reproduction share common molecular changes: upregulation of a neural glycoepitope during morphallaxis in Lumbriculus. AB - Neural morphallaxis is a regenerative process characterized by wide-spread anatomical and physiological changes in an adult nervous system. During segmental regeneration of the annelid worm, Lumbriculus variegatus, neural morphallaxis involved a reorganization of sensory, interneuronal, and motor systems as posterior fragments gained a more anterior body position. A monoclonal antibody, Lan 3-2, which labels a neural glyco-domain in the leech, was reactive in Lumbriculus. In the worm, this antibody labeled neural structures, particularly axonal tracts and giant fiber pathways of the central nervous system. A 60kDa protein, possessing a lumbriculid mannose-rich glycoepitope, was upregulated during neural morphallaxis, peaking in its expression at 3 weeks post-amputation. Peak upregulation of the Lan 3-2 epitope, or the protein possessing it, corresponded to the time of major neurobehavioral plasticity during regeneration. Analyses of asexually reproducing animals also revealed induction of the Lan 3-2 epitope. In this developmental context, Lan 3-2 epitope upregulation was also confined to segments expressing both changes in positional identity and neurobehavioral plasticity, but these molecular and behavioral changes occurred prior to body fragmentation. These results suggest that the lumbriculid Lan 3-2 glycoepitope and proteins that bear them have been co-opted for neural morphallactic programs, induced both in anticipation of reproductive fragmentation and in compensation for injury-induced fragmentation. PMID- 15817229 TI - Reverse genetic analyses of gamete-enriched genes revealed a novel regulator of the cAMP signaling pathway in Dictyostelium discoideum. AB - Sexual development in Dictyostelium discoideum is initiated by the fusion of opposite mating type cells to form zygote giant cells, which subsequently gather and phagocytose surrounding cells for nutrition to form macrocysts. Here we performed the targeting of 24 highly gamete-enriched genes we previously isolated, and successfully generated knockout mutants for 16 genes and RNAi mutants for 20 genes including 6 genes without disruptants. In the knockout mutants of two genes, cell aggregation toward the giant cells was much less extensive and many cells remained around poorly formed macrocysts. We named these genes tmcB and tmcC. Although macrocyst formation of wild type cells was suppressed by the addition of exogenous cAMP, that of knockout mutants of tmcB was much less sensitive. The mRNA level of phosphodiesterase (pde) was higher and that of its inhibitor (pdi) was lower in the latter cells compared to the parental strains during sexual development. Thus, tmcB appeared to be a novel regulator of the cAMP signaling pathway specific to sexual development. Knockout mutants of tmcC were indistinguishable from the wild type cells with respect to the cAMP response, suggesting that this gene is relevant to other processes. PMID- 15817232 TI - Current management of rectal cancer: total mesorectal excision (nerve sparing) technique and clinical outcome. PMID- 15817233 TI - Proctectomy with coloanal anastomosis. PMID- 15817234 TI - Local excision: some reality testing. PMID- 15817235 TI - Surgical management of pelvic malignancy: role of extended abdominoperineal resection/exenteration/abdominal sacral resection. PMID- 15817236 TI - Total pelvic exenteration with distal sacrectomy for fixed recurrent rectal cancer. PMID- 15817237 TI - New treatment concepts for gynecologic pelvic malignancies: neoadjuvant therapies. PMID- 15817238 TI - Cervical cancer: current management of early/late disease. PMID- 15817239 TI - Surgical staging of gynecologic malignancies: the role of laparoscopy and sentinel node technology. PMID- 15817240 TI - Pelvic exenteration of gynecologic malignancy: indications, and technical and reconstructive considerations. PMID- 15817241 TI - Surgical management of prostate cancer: optimizing patient selections and clinical outcome. PMID- 15817242 TI - Bladder cancer--resection/ablation. PMID- 15817243 TI - Laparoscopic lymph node dissection in urologic cancer. PMID- 15817244 TI - Standard reconstruction techniques: techniques of ureteroneocystostomy during urinary diversion. PMID- 15817245 TI - Internal hemipelvectomy for the management of pelvic sarcomas. PMID- 15817246 TI - Extended pelvic resection for sarcoma or visceral tumors invading musculoskeletal pelvis. PMID- 15817247 TI - Image-guided ablative techniques in pelvic malignancies: radiofrequency ablation, cryoablation, microwave ablation. PMID- 15817249 TI - A glimpse of the human field of gastroenterology and applications for veterinary medicine. PMID- 15817250 TI - The ferret gastrointestinal tract and Helicobacter mustelae infection. PMID- 15817251 TI - Gastrointestinal diseases of the ferret. PMID- 15817252 TI - Amphibian gastroenterology. PMID- 15817253 TI - Gastroenterology for the piscine patient. PMID- 15817254 TI - Clinical reptile gastroenterology. PMID- 15817255 TI - Diagnosis and management of Macrorhabdus ornithogaster (formerly megabacteria). PMID- 15817256 TI - Raptor gastroenterology. PMID- 15817257 TI - Disorders of the psittacine gastrointestinal tract. PMID- 15817258 TI - Rabbit gastroenterology. PMID- 15817259 TI - Thalidomide influences the function of macrophages and increases the survival of Plasmodium berghei-infected CBA mice. AB - Malaria remains a major cause of morbidity and mortality in vast areas of the world, mainly due to the severe forms of Plasmodium falciparum infection. The exacerbated immune response, with increased production of TNF and reactive nitrogen and oxygen intermediates, plays a role in the complex pathogenesis of the disease. It is recognised that thalidomide decreases TNF production and may modulate several functions of the immune system. This work evaluated the influence of thalidomide on macrophage functions, and its ability to protect against severe disease. Plasmodium berghei ANKA-infected mice were (n=11) or were not (n=10) intra-gastric treated with thalidomide (150 mg/kg per day), and two other control groups not infected with the parasite were (n=8) or were not (n=10) treated with the drug, and macrophage production of hydrogen peroxide and nitric oxide, and phagocytosis were assessed on the eighth day post-infection. Thalidomide increased the survival time of infected mice, in parallel with a 26.5% increase of the mean of macrophage phagocytic index, and augmented in 13% the mean of the production of hydrogen peroxide and in 45% the mean of nitric oxide production by macrophages related to the non-treated P. berghei-infected mice. Our data indicate that thalidomide improves the outcome of P. berghei ANKA infected CBA mice and suggest that this drug could represent a new alternative to be associated to antimalarial drugs to decrease the morbidity and mortality of severe malaria in non-pregnant individuals. PMID- 15817261 TI - Cellular localization and enzymatic activity of cathepsin B after spinal cord injury in the rat. AB - Mechanical spinal cord injury (SCI) initiates a cascade of pathochemical and pathophysiological events, collectively known as the secondary injury. There has been a long-standing interest in understanding the activation and involvement of proteases in this secondary injury process. Several proteases including the calpains, caspases and matrix metalloproteinases are activated by perturbations to the spinal cord and have been linked to cell death following SCI and in other models of CNS disease and insult. Cathepsin B (Cath B), a potent lysosomal protease, has also been implicated in the pathology of CNS diseases including brain tumors, Alzheimer's disease, amyotrophic lateral sclerosis and stroke. Previously, we reported significant increases in Cath B mRNA and protein expression following contusion-SCI. This characterization of Cath B continues with the experiments reported herein, which were designed to examine Cath B enzymatic activity and cellular localization following contusion-SCI in the rat. Cath B enzymatic activity was significantly increased in the injury epicenter at 5 and 7 days post-injury and was highly correlated with increases in the active forms of the Cath B protein reported earlier. Furthermore, the immunohistochemical analyses revealed that the post-injury increases in expression and enzymatic activity at the injury epicenter were due to the presence of a large and diverse population of inflammatory cells. However, in areas adjacent to the injury epicenter, it appears that parenchymal neurons may also contribute to these increases. Our findings coupled with the documented role of Cath B in other CNS pathologies make this potent protease an attractive candidate for involvement in the tissue destruction associated with the secondary injury cascade following SCI. PMID- 15817260 TI - CD8+-dependent CNS demyelination following ocular infection of mice with a recombinant HSV-1 expressing murine IL-2. AB - Demyelinating diseases comprise a spectrum of immunopathologic syndromes in which myelin, the fatty covering of nerve cell fibers in the brain and spinal cord, is destroyed. In this study, we have shown for the first time that ocular infection of BALB/c mice with a recombinant herpes simplex virus type 1 (HSV-1) expressing IL-2 (HSV-IL-2) results in CNS demyelination as determined by light microscopy and EM. The demyelinated lesions involve periventricular white matter, brain stem, and spinal cord white matter. Demyelination was detected in the CNS of infected mice up to 75 days (the longest time point tested) post HSV-IL-2 infection. In contrast, mice infected with HSV-IFN-gamma or HSV-IL-4, which are identical to HSV-IL-2 but express IFN-gamma or IL-4 instead of IL-2, did not exhibit demyelination. Control mice infected with wild-type HSV-1 or parental virus also remained free of these symptoms. During early times (days 3-7), post infection with HSV-IL-2 virus, a T(H)1 + T(H)2 pattern of cytokines was produced by lymphocytes of infected mice while mice infected with HSV-IFN-gamma or control viruses produced a T(H)1 pattern of cytokine. By day 21 post-infection, all infected groups exhibited a T(H)1 pattern of response. Immunohistochemistry and FACS analyses of infiltrates in the brains and spinal cords of HSV-IL-2-infected mice showed elevations in CD4+ and CD8+ T cells and macrophages. However, T cell depletion studies suggest that only central memory CD8+ T cells are directly involved in the demyelination process, with macrophages being involved through a bystander effect. PMID- 15817262 TI - Serotonergic fiber sprouting to external anal sphincter motoneurons after spinal cord contusion. AB - The present study analyzed the anatomical plasticity of serotonergic immunoreactive projections to external anal sphincter (EAS) motoneurons, and the behavioral plasticity of EAS reflexes, penile erection, and locomotion in rats with spinal contusion injury (SCI) or complete spinal cord transection (TX). Electromyographic activity of the EAS, penile erection latency, and BBB locomotor score exhibited parallel recovery over the 6-week recovery period after contusion SCI. This pattern of recovery was not observed in TX animals. While locomotor scores demonstrated a small increase after TX, erectile and anorectal function remained at abnormal levels established immediately after injury. Serotonergic immunofluorescent (5-HT-IF) staining at the lesion site identified a small number of fibers spared after SCI that may provide a substrate for functional recovery. Pixel density measurements of 5-HT-IF in the vicinity of retrogradely labeled EAS and unlabeled pudendal motoneurons necessary for penile erection provide indirect evidence of serotonergic sprouting that parallels the observed functional recovery in animals with SCI. No 5-HT-IF was detected caudal to the injury site in TX animals. These studies indicate: (1) lumbosacral eliminative and reproductive reflexes provide a valid means of studying the mechanisms of post SCI plasticity; (2) the similar recovery curves suggest similar return of descending control, perhaps through sprouting of descending serotonergic fibers; (3) the observed deficits after TX likely represent the permanent removal of descending inhibition and reflect reorganization of segmental circuitry. PMID- 15817263 TI - Acute and separate modulation of motor and cognitive performance in parkinsonian rats by bilateral stimulation of the subthalamic nucleus. AB - The subthalamic nucleus (STN) is involved in motor and cognitive performance through its key role in the basal ganglia-thalamocortical circuits, but how these different modalities (motor and cognition) are controlled (similar vs. dissimilar) has not yet been elucidated. In the present study, the effects of bilateral STN deep brain stimulation (DBS) on motor and cognitive performance were investigated in a rat model of Parkinson disease (PD). After being trained in a choice reaction time (CRT) task, rats received bilateral injections of 6 hydroxydopamine (6-OHDA) into the striatum. One group of 6-OHDA animals was implanted bilaterally with stimulation electrodes at the level of the STN. Stimulations were performed at 130 Hz (frequency), 60 micros (pulse width), and varying amplitudes of 1, 3, 30, and 150 microA during the CRT task. Finally, rats were sacrificed and the brains processed for staining to determine the dopaminergic lesion (TH immunohistochemistry) and localization of the electrode tip (HE histochemistry). Bilateral 6-OHDA infusion significantly decreased (70%) the number of dopaminergic cells in the substantia nigra pars compacta (SNc) and increased motor time (MT), proportion of premature responding (PR), and reaction time (RT). Bilateral STN stimulation with an amplitude of 3 microA normalized 6 OHDA-induced deficits in PR and RT. Simulation with an amplitude of 30 microA reversed the lesion-induced deficits in MT and RT. Our data show for the first time that bilateral STN stimulation differentially affected the 6-OHDA-induced motor and cognitive deficits. This means that basal ganglia-thalamocortical motor and associative circuits responsible for specific motor and cognitive performance, which are processed through the STN, have unique physiological properties that can acutely and separately be modulated by specific electrical stimuli. PMID- 15817264 TI - Seizure suppression and lack of adenosine A1 receptor desensitization after focal long-term delivery of adenosine by encapsulated myoblasts. AB - Adenosine is an important inhibitory modulator of brain activity. In a previous ex vivo gene therapy approach, local release of adenosine by encapsulated fibroblasts implanted into the vicinity of an epileptic focus, was sufficient to provide transient protection from seizures (Huber, A., Padrun, V., Deglon, N., Aebischer, P., Mohler, H., Boison, D., 2001. Grafts of adenosine-releasing cells suppress seizures in kindling epilepsy. Proc. Natl. Acad. Sci. U. S. A. 98, 7611 7616). Long-term seizure suppression beyond 2 weeks was precluded by limited life expectancy of the encapsulated fibroblasts. To study the feasibility for long term seizure suppression by adenosine releasing brain implants, in the present contribution, mouse C2C12 myoblasts were engineered to release adenosine by genetic inactivation of adenosine kinase. After encapsulation, the myoblasts were grafted into the lateral brain ventricles of epileptic rats kindled in the hippocampus. While seizure activity in animals with wild-type implants remained unaltered, 1 week after grafting all rats with adenosine-releasing implants (n = 25) displayed complete protection from convulsive seizures and a corresponding reduction of afterdischarges in EEG-recordings. The duration of seizure suppression was maintained for a period of 3 weeks in 50% of the animals ranging to a maximum of 8 weeks in one animal. During the course of these experiments, adenosine A1 receptors remained responsive to selective agonists and antagonists indicating a lack of desensitization of A1 receptors after local long-term exposure to adenosine. Furthermore, local release of adenosine did not affect locomotor activity, whereas systemic application of the A1 agonist 2-chloro-N6 cyclopentyladenosine caused strong sedation. Thus, the local release of adenosine by cellular implants provides a feasible option for a potential side-effect free approach for the long-term treatment of focal epilepsies. PMID- 15817265 TI - Lentiviral gene delivery of GDNF into the striatum of R6/2 Huntington mice fails to attenuate behavioral and neuropathological changes. AB - Transgenic R6/2 mice, which express exon 1 of the human mutant Huntington disease gene, develop behavioral and neuropathological changes that bear some resemblance to the human disease. Several studies have shown that elevated glial cell line derived neurotrophic factor (GDNF) levels can exert neuroprotective effects in animal models of Huntington disease that are based on intrastriatal injections of excitotoxins. Therefore, the aim of the present study was to examine whether intrastriatal delivery of the GDNF gene by lentivirus (LV-GDNF) could provide structural and functional protection in R6/2 transgenic mice. Four- to 5-week-old mice were left untreated or alternatively received intrastriatal injections of either LV-GDNF or the same viral vector encoding green fluorescent protein (GFP) (LV-GFP) as a control. During the 4-week follow-up period, there was the expected deterioration in performance of the R6/2 mice in paw clasping, rotarod, and open field tests, and the LV-GDNF treated mice showed no improvement over controls. ELISA showed that the LV-GDNF-injected animals had a significant increase in GDNF level in the striatum, and immunohistochemical analysis revealed that GDNF was also overexpressed in brain regions receiving striatal projections. However, GDNF overexpression had no effect on the neuropathological changes examined. Thus, there were no significant differences in the number of EM-48-positive intraneuronal huntingtin inclusions, number of BrdU-positive cells and size of striatal neuronal cross-sectional area. These results suggest that intrastriatal lentiviral vector transfer of GDNF, performed at 5 weeks of age, does not ameliorate neurological and behavioral impairments in the R6/2 transgenic mice model of HD. Further studies are, however, needed to investigate if GDNF given at earlier time points is beneficial. PMID- 15817266 TI - Dietary supplementation with blueberries, spinach, or spirulina reduces ischemic brain damage. AB - Free radicals are involved in neurodegenerative disorders, such as ischemia and aging. We have previously demonstrated that treatment with diets enriched with blueberry, spinach, or spirulina have been shown to reduce neurodegenerative changes in aged animals. The purpose of this study was to determine if these diets have neuroprotective effects in focal ischemic brain. Adult male Sprague Dawley rats were fed with equal amounts of diets (blueberry, spinach, and spirulina) or with control diet. After 4 weeks of feeding, all animals were anesthetized with chloral hydrate. The right middle cerebral artery was ligated with a 10-O suture for 60 min. The ligature was later removed to allow reperfusional injury. Animals were sacrificed and brains were removed for caspase 3 enzymatic assays and triphenyltetrazolium chloride staining at 8 and 48 h after the onset of reperfusion. A subgroup of animals was used for locomotor behavior and biochemical assays. We found that animals which received blueberry, spinach, or spirulina enriched diets had a significant reduction in the volume of infarction in the cerebral cortex and an increase in post-stroke locomotor activity. There was no difference in blood biochemistry, blood CO2, and electrolyte levels among all groups, suggesting that the protection was not indirectly mediated through the changes in physiological functions. Animals treated with blueberry, spinach, or spirulina had significantly lower caspase-3 activity in the ischemic hemisphere. In conclusion, our data suggest that chronic treatment with blueberry, spinach, or spirulina reduces ischemia/reperfusion induced apoptosis and cerebral infarction. PMID- 15817267 TI - Tumor-induced injury of primary afferent sensory nerve fibers in bone cancer pain. AB - Bone is the most common site of chronic pain in patients with metastatic cancer. What remains unclear are the mechanisms that generate this pain and why bone cancer pain can be so severe and refractory to treatment with opioids. Here we show that following injection and confinement of NCTC 2472 osteolytic tumor cells within the mouse femur, tumor cells sensitize and injure the unmyelinated and myelinated sensory fibers that innervate the marrow and mineralized bone. This tumor-induced injury of sensory nerve fibers is accompanied by an increase in ongoing and movement-evoked pain behaviors, an upregulation of activating transcription factor 3 (ATF3) and galanin by sensory neurons that innervate the tumor-bearing femur, upregulation of glial fibrillary acidic protein (GFAP) and hypertrophy of satellite cells surrounding sensory neuron cell bodies within the ipsilateral dorsal root ganglia (DRG), and macrophage infiltration of the DRG ipsilateral to the tumor-bearing femur. Similar neurochemical changes have been described following peripheral nerve injury and in other non-cancerous neuropathic pain states. Chronic treatment with gabapentin did not influence tumor growth, tumor-induced bone destruction or the tumor-induced neurochemical reorganization that occurs in sensory neurons or the spinal cord, but it did attenuate both ongoing and movement-evoked bone cancer-related pain behaviors. These results suggest that even when the tumor is confined within the bone, a component of bone cancer pain is due to tumor-induced injury to primary afferent nerve fibers that innervate the tumor-bearing bone. Tumor-derived, inflammatory, and neuropathic mechanisms may therefore be simultaneously driving this chronic pain state. PMID- 15817268 TI - Human immunodeficiency virus type 1 transport across the in vitro mouse brain endothelial cell monolayer. AB - Human immunodeficiency virus type 1 (HIV-1) is associated with a neuroinflammatory dementia. Cognitive impairment remains a common complication of late-stage HIV-1 infection. Previous studies have shown that entry of HIV-1 into the central nervous system (CNS) occurs soon after infection. For these reasons, it is important to understand how HIV-1 crosses the BBB. We used primary mouse brain microvessel endothelial cell (MBEC) monolayer models to study interactions between brain endothelial cells and radioactively labeled HIV-1 CL4 (131I-HIV-1), which had been rendered noninfectious with aldithiol, and compared to radioactively labeled bovine serum albumin (131I-BSA or 125I-BSA) and detected HIV-1 on MBEC monolayer with electron microscopic analysis. The permeability of the monolayers to HIV-1 was measured by determining the percent material transported (PMT). Luminal to abluminal PMT of 131I-HIV-1 was 4.65 times greater than that of the much smaller 131I-BSA, showing that the MBEC monolayer is more permeable to HIV-1 than to BSA. Electron microscopy showed that HIV-1 was transported through a trans-cellular pathway from luminal side to basolateral space with some virus associated with the nucleus. Unlabeled HIV-1 did not affect the transport of 131I-HIV-1 or break down the MBEC monolayer. Wheatgerm agglutinin (WGA) increased 131I-HIV-1 penetration across the MBEC monolayer, consistent with absorptive endocytosis as the mechanism for HIV-1 penetration. The enhanced transport of HIV-1 was unidirectional, as the abluminal to luminal PMT of 131I-HIV-1 was not different from that of BSA nor enhanced by WGA. Characterization of the radioactivity transported from the luminal to abluminal chamber on Sepharose 4B-200 columns showed the transported radioactivity represented intact virus. MBEC monolayers preloaded from the luminal surface with 131I-HIV-1 showed most of the virus was retained by the endothelial cells, while the remainder was effluxed mainly to the luminal surface. MBEC monolayers preloaded from the abluminal surface with 131I-HIV-1 retained little virus and most of the virus was effluxed mainly to the abluminal surface. In conclusion, cell-free, intact 131I-HIV-1 crossed brain endothelial cell monolayers unidirectionally in the luminal to abluminal direction through an adsorptive endocytotic pathway. HIV-1 taken up from luminal side by monolayers of brain endothelial cells was mainly released to the luminal side. HIV-1 efflux mechanisms are different from influx mechanisms. PMID- 15817269 TI - Effect of GABA(A) receptor stimulation in the subthalamic nucleus on motor deficits induced by nigrostriatal lesions in the rat. AB - Inhibition of the subthalamic nucleus by lesions or GABAergic agonists improves motor symptoms in monkeys or humans with a loss of nigrostriatal dopaminergic neurons, a characteristic of Parkinson's disease. In rats, nigrostriatal lesions induce deficits in a variety of motor tests that are ameliorated by dopaminergic agonists. However, the validity of these tests to predict the beneficial effects of subthalamic inhibition is not known. We have examined the effects of an intrasubthalamic injection of the GABA(A) receptor agonist muscimol (0.1 microg/0.1 microL) in intact rats and in rats with a unilateral nigrostriatal lesion. Muscimol induced a mild ipsiversive rotation in sham-operated (control) rats and blocked contraversive rotations induced by apomorphine in lesioned rats. In addition, in the cylinder test of limb use asymmetry, muscimol decreased the ipsilateral bias after lesion without inducing any significant effect in sham operated controls. In the forced-step test, however, 0.1 microg (but not 0.01 microg) of muscimol into the subthalamic nucleus induced a behavioral bias by markedly decreasing the number of adjusting steps of the contralateral limb in control rats, similar to the effect of a nigrostriatal lesion. Neither dose improved performance in this test in rats with lesions, and the higher dose exacerbated the deficit. The data support a beneficial role of stimulating subthalamic GABA(A) receptors for akinesia but also reveal negative behavioral effects of this treatment and suggest that the cylinder and forced-step tests measure different aspects of behavioral deficits after dopaminergic lesions. PMID- 15817270 TI - Intraventricular injections of tachykinin NK3 receptor agonist reduce the gain of the baroreflex in unrestrained rats. AB - The tachykinin neuropeptides acting at NK3 receptors affect mean arterial pressure (MAP) through both neuroendocrine and neural mechanisms. NK3 receptors are found in brainstem nuclei that mediate the baroreflex, but the effects of NK3 receptor stimulation on baroreflex function are unknown. The present study tests the effects of intraventricular injections of senktide, a selective NK3 receptor agonist, on the sensitivity of the baroreflex in three stains of rats: Charles River Laboratory, Long-Evans, and Brattleboro rats, which lack the ability to synthesize vasopressin. Rats with lateral ventricle cannulas were administered injections of isotonic saline, 100 ng, or 200 ng senktide, and 5 min later arterial baroreceptor-heart rate (HR) function was examined by constructing full range blood pressure-HR curves using alternating doses (5-20 microg kg min) of phenylephrine and nitroprusside to raise and decrease blood pressure approximately 50 mm Hg over a period of 1 min, respectively. Intraventricular injections of 200 ng senktide had no significant effect on baseline MAP, but significantly decreased the gain of the baroreflex in all three rat strains whereas the 100 ng dose had no effect on the baroreflex. These results show that NK3 receptor stimulation modulates the baroreflex that is independent of any action of vasopressin. PMID- 15817271 TI - Regional distribution of fluoro-jade B staining in the hippocampus following traumatic brain injury. AB - Fluoro-Jade B (FJB) is an anionic fluorescein derivative that has been reported to specifically stain degenerating neurons. We were interested in applying FJB staining in a well-characterized model of traumatic brain injury (TBI) in order to estimate the total number of neurons in different regions of the hippocampus that die after a mild or moderate injury. Rats were subjected to a mild or moderate unilateral cortical contusion (1.0- or 1.5-mm displacement from the cortical surface) with a controlled cortical impacting device. Animals were allowed to survive for 1, 2, or 7 days and the total number of FJB-positive neurons in hippocampal areas CA1, CA3, and the dentate gyrus granule layer was estimated using sterological methods. The region that had the highest number of FJP-positive neurons after TBI was the dentate gyrus. In both 1- and 1.5-mm injuries, FJB-positive granule cells were observed throughout the rostro-caudal extent of the dentate. In contrast, labeled pyramidal neurons of area CA3 were most numerous after the 1.5-mm injury. The area that had the fewest number of FJB labeled cells was area CA1 with only scattered neurons seen in the 1.5-mm group. In both injury groups and in all hippocampal regions, more FJB-positive neurons were seen at the earlier times post injury (1 and 2 days) than at 7 days. FJB appears to be a reliable marker for neuronal vulnerability following TBI. PMID- 15817272 TI - Apomorphine-induced alterations in striatal and substantia nigra pars reticulata glutamate following unilateral loss of striatal dopamine. AB - We have reported time-dependent changes in extracellular glutamate within the striatum at 1 and 3 months following a unilateral lesion of the nigrostriatal pathway using the neurotoxin, 6-hydroxydopamine (6-OHDA) (Meshul, C.K., Emre, N., Nakamura, C.M., Allen, C., Donohue, M.K., Buckman, J.F., 1999. Time-dependent changes in striatal glutamate synapses following a 6-hydroxydopamine lesion. Neurosci. 88, 1-16.). The aim of the present study was to determine the effects of such a lesion on glutamate within the substantia nigra pars reticulata (SN-PR) and the effect of subchronic administration of the dopamine D-1/D-2 agonist, apomorphine, on extracellular glutamate within both the striatum and the SN-PR using in vivo microdialysis. One month after the lesion, there is an increase in extracellular glutamate within the striatum and apomorphine treatment leads to a further increase. Within the SN-PR, a loss of striatal dopamine leads to a decrease in extracellular glutamate, while apomorphine treatment leads to a further decrease in nigral glutamate. Three months after a 6-OHDA lesion, there is a decrease in extracellular striatal glutamate, with apomorphine administration leading to essentially no further change in glutamate. The loss of striatal dopamine increased extracellular glutamate within the SN-PR while apomorphine administration resulted in a decrease in extracellular glutamate back to the value observed in the control group. The data suggests that the increase in striatal glutamate 1 month following a 6-OHDA lesion alone or following subchronic apomorphine is consistent with the hypothesis that a decrease in glutamate within the SN-PR leads to activation of the thalamo-cortico-striatal pathway. The decrease in striatal glutamate 3 months after a nigrostriatal lesion is also consistent with the observed increase in extracellular glutamate within the SN-PR, thus leading to a decrease in output of the thalamo-cortico-striatal pathway. PMID- 15817273 TI - Calbindin distribution in male, female and lactating rat pituitary. AB - Calcium binding proteins such as calbindin and calretinin have been studied in the pituitary gland, but information on them is still incomplete. To investigate the localization, distribution and role of calbindin in the pituitary, several antibodies to calbindin and to other pituitary markers, such as calretinin and tyrosine-hydroxylase, have been used in male, female and lactating rats. Calbindin has not been localised to a specific endocrine cell population unlike calretinin in the thyrotrophs. There was occasional localization in somatotrophs, thyrotrophs and luteotrophs, but not in corticotrophs or lactotrophs. However, there are sex differences in the expression of this protein as the number of calbindin-immunoreactive cells is higher in the male than in the female pituitary. Furthermore, the number of calbindin containing cells, not lactotrophs, increases in lactating rats and decreases after removal of the pups. It is concluded that calbindin expression may be altered by physiological and endocrine events such lactation, even though it is still unclear why the protein is not related to a specific cell type. The simultaneous use of monoclonal and polyclonal antisera to calbindin revealed that the rabbit antibody recognizes nuclear and cytoplasmic calbindin, while the monoclonal one binds only to the cytoplasmic calbindin. The suggestion is that calbindin may have a secondary role that is not simply to bind calcium. PMID- 15817274 TI - The effect of hypoxic-ischemic brain injury in perinatal rats on the abundance and proteolysis of brevican and NG2. AB - Oligodendrocyte (OL) progenitor cells are particularly susceptible to perinatal hypoxia/ischemia (H-I) resulting in decreased myelination and attenuated development of white matter fiber tracts. Brevican is an aggregating chondroitin sulfate proteoglycan (CSPG) secreted by OLs and their progenitors prior to and during active developmental myelination whereas neuron-glia antigen 2 (NG2) is a transmembrane CSPG produced by early OL progenitors. Although both proteoglycans are associated with maturation of OLs, it is not known if they are altered by H-I brain injury in the neonate. We have therefore examined the time course of changes in brevican and NG2 abundance and proteolysis in the neonatal rat hippocampus after H-I. In a standard H-I model of unilateral carotid artery ligation and exposure to hypoxia, a cavitary infarct involving the ipsilateral parietal and temporal regions of cerebral cortex, hippocampus, and striatum of most rat pups was clearly evident 4 days after H-I. The abundance of total extractable brevican was markedly reduced in the ipsilateral hippocampus at 1 and 14 days after H-I (relative to the contralateral side). At these times, the total G1 proteolytic fragment of brevican was lower in the ipsilateral hippocampus and the level of a protease-generated brevican fragment was significantly diminished in the OL-rich hippocampal fimbria. Hippocampal NG2 levels were also lower at 1 and 4 days after H-I, but were not different from the contralateral side at 14 days. Since brevican, brevican G1 fragment, and NG2 loss occur around the time of progressive cell death and the appearance of the infarct, it may be that H-I rapidly induces a cellular response that actively depletes these proteoglycans from the hippocampal matrix. While the mechanism of this loss is unclear, it would appear to be an early event in the process that could be involved in apoptotic cell death and/or tissue injury. PMID- 15817275 TI - Simvastatin treatment does not protect retinal ganglion cells from degeneration in a rat model of autoimmune optic neuritis. AB - In patients with multiple sclerosis (MS), non-remitting deficits are mainly caused by axonal and neuronal damage. We demonstrated previously that myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis in rats provokes severe axonal and neuronal injury even before clinical manifestation of the disease. In our present study, we investigated effects of simvastatin treatment on degeneration of retinal ganglion cell (RGC) bodies as well as their axons during MOG-induced optic neuritis. Electrophysiological functions of optic nerves and RGCs were analyzed in vivo. Although neuroprotective effects of simvastatin have been demonstrated before in other experimental settings, we did not observe an increase in RGC survival nor an improvement of visual functions. As we could not reproduce the anti inflammatory effects that were observed under statin therapy in other EAE models, we hypothesize that patients suffering from optic neuritis might not take advantage of simvastatin applications. PMID- 15817276 TI - MRI guidance improves accuracy of stereotaxic targeting for cell transplantation in parkinsonian monkeys. AB - Accuracy of targeting is critical for the success of cell transplantation in the central nervous system. We compared the accuracy of conventional atlas-guided stereotaxis to magnetic resonance imaging (MRI)-guided stereotaxic targeting in various basal ganglia nuclei in parkinsonian monkeys. 28 monkeys underwent unilateral striatal transplantation. High-resolution 3D MR images of the brain were used in 15 monkeys fitted with a MRI-compatible stereotaxic frame for target localization. This was immediately followed by cranial surgery with the frame "in situ". 13 additional monkeys underwent stereotaxic atlas-guided cranial surgery for placement of cell transplants. Following extensive behavioral testing and microelectrode recordings, all animals were perfused. The brains were sectioned coronally and stained to determine the morphology of needle tracts as an accuracy measure of stereotaxic placements. MRI-guided stereotaxy was completely accurate in 80% as compared to 38.5% in atlas-guided stereotaxis. The chance of missing a target completely was as high as 38.5% in atlas-guided stereotaxis, which was reduced to 6.67% when MRI was used for guidance. Targeting error occurred mostly in the anterior caudate and posterior putamen as against better accuracy in the anterior putamen. These results suggest that accuracy of stereotaxic unilateral cranial targeting into the putamen and the caudate in monkeys can be improved with high-resolution 3D MR imaging. PMID- 15817277 TI - Treadmill training ameliorates dopamine loss but not behavioral deficits in hemi parkinsonian rats. AB - The purpose of this study was to investigate whether locomotor training could ameliorate neurochemical changes and behavioral deficits in the 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease. It has been recently demonstrated that forelimb motor training, or brief treadmill training, can attenuate dopamine loss and some deficits in forelimb usage in this animal model. Nevertheless, it is not known whether locomotor training could result in an amelioration of locomotor deficits. Rats were lesioned with 6-OHDA injected intracerebrally and randomly assigned to one of 3 groups: early treadmill trained, late treadmill trained and untrained. Animals in the early trained group underwent 2 x 20 min treadmill sessions daily for 30 days, beginning 24 h after 6-OHDA injection. Late trained animals underwent the same training regime beginning 7 days post-injection. All animals were assessed on their abilities to perform several behavioral tasks designed to test locomotor and forelimb movement abilities prior to 6-OHDA injection and at 3 and 6 weeks post-injection. Treadmill training resulted in the attenuation of dopamine depletion in the striatum compared to non-treadmill trained animals, as measured by in vivo apomorphine-induced rotations and post mortem dopamine analysis. Nevertheless, treadmill training produced essentially no difference in behavioral deficits on most tests compared to untrained animals. We discuss the possible reasons for the discrepancies with previous studies, including differences in lesioning, training regimes and methods of behavioral assessment. We conclude that treadmill training does not ameliorate locomotor deficits in the 6-OHDA model of Parkinson's disease, even though this same training results in attenuation of dopamine loss in the striatum. PMID- 15817278 TI - Stereologic analysis of hippocampal Alzheimer's disease pathology in the oldest old: evidence for sparing of the entorhinal cortex and CA1 field. AB - Several neuropathologic analyses postulate that Alzheimer disease (AD) in the oldest-old is associated with substantial neurofibrillary tangle (NFT) formation in the CA fields of the hippocampus and neuronal loss confined to the entorhinal cortex. All of these studies have measured densities, rather than absolute numbers, and most do not take into account the potential interaction between the above pathological hallmarks in a global multivariate analysis. We present here a stereologic analysis of AD-related pathology in 12 oldest-old individuals including a complete assessment of total NFT, neuron numbers and amyloid volume in entorhinal cortex, CA fields, and dentate gyrus. The progression of NFT numbers and amyloid volume across the different Clinical Dementia Rating (CDR) groups was significantly slower in these cases compared to previously reported younger cases. Although patients with mild and moderate dementia showed significantly lower mean neuron numbers compared to CDR 0-0.5 cases, there was a marked overlap in individual values among CDR groups. A modest proportion of the variability in CDR scores was explained by NFT numbers in the CA2 field (18.1%) and the dentate gyrus (17.3%). In contrast, neither Nissl-stained neuron numbers nor total amyloid volume in the areas studied significantly predicted cognitive status. These data indicate that the occurrence and progression of AD-related pathologic changes are not an unavoidable consequence of aging. They also suggest that dementia in extreme aging depends more on the damage of hippocampal subdivisions commonly less affected than on severe NFT formation and neuronal loss in the CA1 field and entorhinal cortex. PMID- 15817279 TI - Development of interlaminar astroglial processes in the cerebral cortex of control and Down's syndrome human cases. AB - Glial cytoarchitecture in human cerebral cortex is constituted by two overlapping layouts: the (general mammalian) "glial syncytium" and the (primate-specific) "interlaminar glial palisade" (IGP) composed by astroglial cells, with long, radial processes that traverse several supragranular layers. In this study, the emergence and early organization of the IGP was analyzed using immunocytochemical procedures in postmortem infantile human control and age matched, Down's syndrome (DS) cases. In control cases, first signs of a radial array of unbranched astroglial processes were apparent at the end of the period of "physiological astrocytosis" (20-40 days of postnatal life), and its general profile (except perhaps the density of cell processes) reached the adult-like configuration by the second month of life. The initial organization of the IGP was similar in control and DS cases, although a breakdown in DS became manifest by the first year of age, or earlier, albeit with individual variations. These changes tended to evolve in a "mosaic" fashion and included partial disruption of the palisade, or persistence of the "physiological astrocytosis". These observations were compared against samples from elder DS cases with an Alzheimer's type of dementia (AtD). Collectively, results suggest that DS also involves astroglial alterations during early stages of brain development, and that those changes progress with age, until an AtD ensues during adult life. PMID- 15817280 TI - Permeability of the blood-brain barrier to HIV-1 Tat. AB - Infection with human immunodeficiency virus-1 (HIV-1) is associated with dysfunctions of the central nervous system (CNS). HIV-1 induces its effects on the CNS by a variety of mechanisms, including by shedding the neurotoxic viral proteins such as gp120 and Tat. Both HIV-1 and gp120 have been shown to cross the blood-brain barrier (BBB). It is has not been determined, however, whether blood borne Tat can cross the BBB. Here, we found that Tat crosses the BBB by a nonsaturable mechanism with a unidirectional influx rate of about 0.490 microl/g/min. About 0.126% of an intravenous dose of Tat enters each g of brain. Radioactively labeled albumin injected simultaneously did not cross the BBB. The hypothalamus, occipital cortex, and hippocampus were the regions of the brain most permeable to Tat. Nonsaturable brain-to-blood efflux also occurred, most likely with reabsorption into the blood of the cerebrospinal fluid. In conclusion, we found that Tat crossed the BBB bidirectionally. Such permeability could provide a mechanism by which Tat produced on one side of the BBB could affect neural or immune function on the other side. PMID- 15817281 TI - Are tuberomammillary histaminergic neurons involved in CO2-mediated arousal? AB - An increase in arousal in response to hypercapnia [elevated arterial PCO2 (partial pressure of CO2) levels] during awake or sleep states is an important component of mechanisms designed to maintain acid-base homeostasis. Since central histaminergic neurons are crucial for maintaining waking states and vigilance, a nonresponsive or dysfunctional histaminergic system could contribute to the lack of arousal in response to hypercapnia in some sleep-related disorders [e.g., sudden infant death syndrome (SIDS) and Ondine's curse]. Therefore, the present study attempted to determine if histaminergic neurons display functional responses to acute exposure to hypercapnic gas (i.e., gas with elevated CO2 concentrations). Healthy adult male rats were placed in flow cages during the light cycle, or inactive phase, and exposed to either atmospheric air or to environmental CO2 concentrations increasing from baseline up to 20% CO2 over a 5 min period. The expression of the protein product of the immediate-early gene c fos was used as a measure of functional cellular responses within subpopulations of histaminergic neurons. Among the histaminergic subgroups (E1-E5), only the ventral tuberommamillary nucleus (VTMn)/E2 cell group showed significant increases in c-Fos expression following brief exposure to hypercapnic gas. These data are consistent with the hypothesis that histaminergic neuronal cell groups are heterogeneous and are involved in physiological and/or behavioral responses to acute hypercapnic challenge, potentially increasing vigilance during active waking and awakening from sleep during hypercapnic states. PMID- 15817282 TI - Modulation of parvalbumin expression in the motor cortex of parkinsonian rats. AB - Dopamine deficiency in Parkinson's disease leads to numerous molecular changes in basal ganglia. However, the consequences of these changes on the motor cortex remain unclear. Here we show that the immunoreactivity of parvalbumin, which is expressed in GABAergic interneurons, increases in the primary motor cortex of parkinsonian rats. This increase can be reversed by a subsequent lesion of the subthalamic nucleus. These results suggest that dopamine deficiency induces reversible changes in GABAergic cortical cells, which might be linked with parkinsonian symptoms. PMID- 15817283 TI - Circadian rhythms. Preface. PMID- 15817284 TI - Analysis of circadian rhythms in Neurospora: overview of assays and genetic and molecular biological manipulation. AB - The eukaryotic filamentous fungus Neurospora crassa is a tractable model system that has provided numerous insights into the molecular basis of circadian rhythms. In the core circadian clock feedback loop, WC-1 and WC-2 interact via PAS domains to heterodimerize, and this complex acts both as the circadian photoreceptor and, in the dark, as a transcription factor that promotes the expression of the frq gene. In the negative step of the loop, dimers of FRQ feed back to block the activity of the WC-1/WC-2 complex (WCC) and, in a positive step, to promote the synthesis of WC-1. Several kinases phosphorylate FRQ, leading to its ubiquitination and turnover, releasing the WC-1/WC-2 dimer to reactivate frq expression and restart the circadian cycle. Light and temperature entrainment of the clock arise from rapid light induction of frq expression and from the effect of elevated temperatures in driving higher levels of FRQ. Noncircadian candidate slave oscillators, termed FRQ-less oscillators (FLOs), have been described, each of which appears to regulate aspects of Neurospora growth or development. Overall, the core FRQ/WCC feedback loop coordinates the circadian system by regulating downstream clock-controlled genes either directly or via regulation of driven FLOs. This article provides a brief synopsis of the system and describes current assays for the Neurospora clock. Methods for genetic and molecular manipulation of the core clock are summarized, and accompanying chapters address more specifically aspects of photobiology and output. PMID- 15817285 TI - Circadian genetics in the model higher plant, Arabidopsis thaliana. AB - In recent decades, most research on the circadian rhythms of higher plants has been driven by molecular genetics. A wide variety of experimental approaches have discovered mutants in the plant circadian clock, yet the screens are far from saturated and there must still be important clock-related genes to identify. Direct methods to screen for circadian mutants include the original assay of rhythmic luminescence from promoter:luciferase constructs in planta or a recently developed assay based on stomatal rhythms. Mutants found through simpler screens of processes only partially controlled by the clock are still identifying novel and interesting circadian phenotypes when their rhythms are tested, while the sequenced genome and the large range of mutant stocks available have made reverse genetics increasingly powerful. PMID- 15817286 TI - Genetic screens for clock mutants in Drosophila. AB - The isolation and analysis of mutant flies (Drosophila melanogaster) with altered circadian rhythms have led to an understanding of circadian rhythms at the molecular level. This molecular mechanism elucidated in fruit flies is similar to the mechanism of the human circadian clock, which confers 24-h rhythmicity to our sleep/wake behavior, as well as to many other aspects of our cellular and organismal physiology. In fruit flies, genes can be mutated to abolish circadian rhythms (i.e., produce arrhythmia) or alter the period of the circadian rhythm; these genes encode key components of the circadian oscillator mechanism. Other mutations have identified components of the input pathways (by which light and temperature synchronize the circadian clock to environmental cycles) or output pathways (which connect the circadian oscillator to the physiological response). Mutations in genes are typically generated by chemical mutagenesis or mutagenesis with transposable elements. Flies with mutagenized chromosomes are processed in a series of genetic crosses, which allow specific chromosomes to be screened for semidominant mutations, recessive mutations, enhancer/suppressor mutations, or genes that can be overexpressed to alter circadian rhythms. Circadian phenotypes, which are assayed to identify mutants, include eclosion (emergence of the adult from the pupal case), locomotor activity (similar to human sleep?wake behavior), and circadian oscillations of gene expression. It is argued that screens for new circadian genes will continue to reveal novel components of the circadian mechanism. PMID- 15817287 TI - Systems approaches to biological rhythms in Drosophila. AB - The chronobiological system of Drosophila is considered from the perspective of rhythm-regulated genes. These factors are enumerated and discussed not so much in terms of how the gene products are thought to act on behalf of circadian-clock mechanisms, but with special emphasis on where these molecules are manufactured within the organism. Therefore, with respect to several such cell and tissue types in the fly head, what is the "systems meaning" of a given structure's function insofar as regulation of rest-activity cycles is concerned? (Systematic oscillation of daily behavior is the principal overt phenotype analyzed in studies of Drosophila chronobiology). In turn, how do the several separate sets of clock-gene-expressing cells interact--or in some cases act in parallel--such that intricacies of the fly's sleep-wake cycles are mediated? Studying Drosophila chrono-genetics as a system-based endeavor also encompasses the fact that rhythm related genes generate their products in many tissues beyond neural ones and during all stages of the life cycle. What, then, is the meaning of these widespread gene-expression patterns? This question is addressed with regard to circadian rhythms outside the behavioral arena, by considering other kinds of temporally based behaviors, and by contemplating how broadly systemic expression of rhythm-related genes connects with even more pleiotropic features of Drosophila biology. Thus, chronobiologically connected factors functioning within this insect comprise an increasingly salient example of gene versatility--multi faceted usages of, and complex interactions among, entities that set up an organism's overall wherewithal to form and function. A corollary is that studying Drosophila development and adult-fly actions, even when limited to analysis of rhythm-systems phenomena, involves many of the animal's tissues and phenotypic capacities. It follows that such chronobiological experiments are technically demanding, including the necessity for investigators to possess wide-ranging expertise. Therefore, this chapter includes several different kinds of Methods set-asides. These techniques primers necessarily lack comprehensiveness, but they include certain discursive passages about why a given method can or should be applied and concerning real-world applicability of the pertinent rhythm-related technologies. PMID- 15817288 TI - Analysis of circadian rhythms in zebrafish. AB - The zebrafish probably constitutes the best animal system to study the complexity of the circadian clock machinery and the influence that light has on it. The possibilities of producing transgenic fishes, to establish light-responsive cultured cells, and to directly explore light phototransduction on single clock cells are all remarkable features of this circadian system. This article describes some of the most useful methodologies to analyze the behavioral, cellular, and molecular aspects of the zebrafish circadian clock system. PMID- 15817289 TI - Genetic manipulation of circadian rhythms in Xenopus. AB - Xenopus laevis retina is an important experimental model system for the study of circadian oscillator mechanisms, as light input pathways, central oscillator mechanisms, and multiple output pathways are all contained within this tissue. These retinas continue to exhibit robust circadian rhythms even after being maintained in culture for many days. The usefulness of this system has been improved even further by the development of a technique for simple genetic manipulation of these animals, which is complemented by expanded genomics resources (Xenopus genome project, microarray, etc.). By taking advantage of the transgenic technique in Xenopus described in this article, many types of analysis can be done on the primary transgenic animals within a couple of weeks after transgenesis. The availability of many cell-type-specific promoters and well characterized cell types within the Xenopus retina provides the advantage of cell specific modification of clock function using this method; in other words, contributions of different cell types within the circadian system can be analyzed independently by "molecular dissociation" of these cells. This article describes both how this transgenic technique is useful and various considerations that should be taken into account when these types of experiments are planned and interpreted. Application of these new techniques to studies of clock function provide an opportunity to rapidly assess gene expression and?or function in the context of the intact retina. PMID- 15817290 TI - Forward genetic screens to identify circadian rhythm mutants in mice. AB - This article describes the methods and techniques used to produce mutagenized mice to conduct high-throughput forward genetic screens for circadian rhythm mutants in the mouse. In particular, we outline methods to safely prepare and administer the chemical mutagen N-nitroso-N-ethylurea (ENU) to mice. We also discuss the importance of selecting mouse strain and outline breeding strategies, logistics, and throughput to produce these mutant mice. Finally, we discuss the breeding strategies that we use to confirm mutation heritability. PMID- 15817291 TI - Methods to record circadian rhythm wheel running activity in mice. AB - Forward genetic approaches (phenotype to gene) are powerful methods to identify mouse circadian clock components. The success of these approaches, however, is highly dependent on the quality of the phenotype--specifically, the ability to measure circadian rhythms in individual mice. This article outlines the factors necessary to measure mouse circadian rhythms, including choice of mouse strain, facilities and equipment design and construction, experimental design, high throughput methods, and finally methods for data analysis. PMID- 15817292 TI - Genetic approaches to human behavior. AB - Tremendous progress in the field of human genetics has made a major impact over the last two decades into our understanding of many Mendelian disorders affecting humans. It is much more difficult to approach the genetics of complex disorders and is particularly challenging in the case of dissecting genetics of the wide range of behavioral variation in the general population. Knowledge of the biology and genetics of Mendelian traits is beginning to inform studies in complex genetics. Furthermore, the convergence of this growing knowledge with increasingly powerful new tools now put complex genetics of behavior within reach. The bottleneck in such studies and the greatest challenge to investigators is the rigorous phenotyping of human research subjects. PMID- 15817293 TI - Enhanced phenotyping of complex traits with a circadian clock model. AB - Models of biological systems are increasingly used to generate and test predictions in silico. This article explores the basic workings of a multifeedback network model of a circadian clock. In a series of in silico experiments, we investigated the influence of the number of feedbacks by adding and removing one or more. We further explore the possibilities of testing in silico models in classic "circadian" protocols. In addition, we performed an in silico mutagenesis screen (by altering parameters throughout the network), creating a library of mutants (based on "phenotype," not "genotype"), and subjected them to a variety of straightforward "circadian" protocols. The results of this mutant "taxonomy" are surprising. While most mutants can be identified (separated) using a limited set of experimental protocols, some resist such a separation, even when "mutations" are at vastly different locations within the complex model. Furthermore, some protocols distinguish similar alleles of the same component, which would be counterproductive. The described taxonomy invites experimental verification, in vivo, and may ultimately streamline genotyping of complex traits, which may have been based previously on imprecise phenotypes. PMID- 15817294 TI - Real-time reporting of circadian-regulated gene expression by luciferase imaging in plants and mammalian cells. AB - Luciferase enzymes have been used as reporters of circadian rhythms in organisms as diverse as cyanobacteria, plants, fruit flies, and mice. This article details methodology for real-time reporting of circadian-regulated gene expression by imaging of luciferase bioluminescence in plants and mammalian cells. PMID- 15817295 TI - Real-time luminescence reporting of circadian gene expression in mammals. AB - Luminescence reporters have been used successfully in studies of circadian rhythms. Real-time measurements of circadian variations in gene expression were made in living cells, cultured tissues, and whole organisms. Because this technique is relatively easy and continuous noninvasive measurement from tissue cultures allows for a drastic reduction in the number of experimental animals, we believe this method will become a common technique for studying circadian rhythms. Using a multichannel recording apparatus, it may also become a powerful tool for the discovery of new drugs. In the past, measurements were done using hand-made apparatuses or by modifying commercially available equipment. We, along with other investigators, have developed user-friendly equipment for performing circadian rhythms experiments, and these systems are now available commercially. This article describes the use of luminescence reporters in circadian research and provides detailed methods used in these experiments. One of our goals in this article is to reduce experimental variability in different laboratories by proposing standard protocols. PMID- 15817296 TI - Transgenic cAMP response element reporter flies for monitoring circadian rhythms. AB - The cAMP response Element (CRE)-binding protein (CREB) is involved in many adaptive behaviors, including circadian rhythms. In order to assess CREB activity in vivo, we made transgenic flies carrying a CRE-luciferase reporter and showed that this reporter is CRE and dCREB2 responsive. dCREB2 is the Drosophila homolog of mammalian CREB?CREM. The transgenic luciferase activity cycles with a 24-h periodicity, suggesting that dCREB2 and period are somehow linked. The CRE luciferase reporter is a useful monitor of circadian activity, and mutations can be found that affect its periodicity, baseline activity, or amplitude. Analysis of such mutations should reveal information about how particular genes affect the molecular machinery of circadian cycling and how different genes affect the activity of dCREB2. PMID- 15817297 TI - Analysis of circadian output rhythms of gene expression in Neurospora and mammalian cells in culture. AB - The true biology of chronobiology lies in the spectrum of processes that are controlled by the circadian clock. Although this biology plays out at the level of the whole organism, it derives, finally, from clock-driven changes in physiology, and frequently in gene expression, that occur at the level of individual cells. For this reason, analysis of gene expression rhythms measured in cell culture or in organisms that elaborate only a few cell types provides insights not possible in multicellular organisms. In this context we have used mammalian fibroblasts in culture as well as the eukaryotic filamentous fungus Neurospora crassa to study circadian output, in particular the output rhythms in gene expression that underlie so much of circadian biology. Each cell type has its own advantages: Data from mammalian cells are obviously immediately pertinent to animal cell rhythms, but the system allows little genetics and only limited amounts of material can be collected. Alternatively, Neurospora allows genetic and molecular analyses and is useful for developing concepts and models of output that can be examined in other contexts. This methods article focuses on these two systems for analysis, providing an overview of how control is presently viewed followed by current methods for its analysis. PMID- 15817298 TI - Molecular and statistical tools for circadian transcript profiling. AB - This article describes methods used to evaluate mRNA expression patterns on microarrays and their application in circadian biology. With the intention of complementing rather than duplicating the existing literature, particular emphasis is placed on experimental design, data analysis techniques, and independent verification. Both comparative and temporal study designs are discussed, and their use in circadian research is illustrated with examples. Data analysis methods to assess periodic components in time series data are outlined in detail. PMID- 15817299 TI - RNA profiling in circadian biology. AB - DNA arrays have become indispensable tools in the study of transcriptional output of the circadian clock and have enabled an increase in the number of outputs from tens to thousands. However, despite their widespread use, many challenges exist in accurately and sensitively identifying all transcriptional targets of the clock. This article discusses aspects of experimental design, including RNA amplification strategies, data condensation methods, and other aspects that impact the use of these tools for the study of circadian biology. PMID- 15817300 TI - Analysis of posttranslational regulations in the Neurospora circadian clock. AB - Posttranslational modification of circadian clock proteins by phosphorylation is an essential regulatory process in the control of eukaryotic circadian clocks. In the Neurospora circadian clock, the key clock protein FREQUENCY (FRQ) is progressively phosphorylated. The phosphorylation of FRQ is regulated by both kinases and phosphatases, and the phosphorylation is important for regulating FRQ stability and its function in the circadian negative feedback loop. The degradation of FRQ is mediated by the ubiquitin/proteasome pathway. This article discusses posttranslational regulations of the Neurospora clock and describes the methods used in the studies of FRQ phosphorylation, FRQ kinases and phosphatases, and FRQ degradation. PMID- 15817301 TI - Analyzing the degradation of PERIOD protein by the ubiquitin-proteasome pathway in cultured Drosophila cells. AB - Time-of-day specific changes in the levels of key clock proteins are critical for the normal progression of circadian pacemakers. Evidence indicates a major role for the ubiquitin-proteasome pathway (UPP) in the temporal control of clock protein stability. A conserved feature of animal clocks is that PERIOD (PER) proteins undergo daily rhythms in abundance. The stability of PER proteins is regulated by differential phosphorylation, whereby hyperphosphorylated isoforms are selectively degraded by the UPP. The use of transformed stable cell lines has been instrumental in advancing our understanding of the mechanisms underlying the intersection of the UPP and clock protein metabolism. This article describes several standard methodologies used to analyze the UPP-mediated degradation of Drosophila PER (dPER) expressed in cultured Drosophila cells (Ko et al., 2002). Although this article focuses on dPER as a case study, general issues are discussed that should have broad application to other cell culture-based systems and clock proteins. For example, we discuss (i) advantages?disadvantages of cultured cells, (ii) types of expression vectors and "peptide tags" for recombinant protein production and surveillance, and (iii) standard approaches to determine whether a protein of interest is modified by ubiquitin and degraded by the proteasome. Prior to the discussion on methodologies, the article provides a brief overview of diverse strategies by which clock proteins in a variety of systems are regulated by the UPP. PMID- 15817302 TI - Casein kinase I in the mammalian circadian clock. AB - The circadian clock is characterized by daily fluctuations in gene expression, protein abundance, and posttranslational modification of regulatory proteins. The Drosophila PERIOD (dPER) protein is phosphorylated by the serine?threonine protein kinase, DOUBLETIME (DBT). Similarly, the murine PERIOD proteins, mPER1 and mPER2, are phosphorylated by casein kinase I epsilon (CKI), the mammalian homolog of DBT. CKIepsilon also phosphorylates and partially activates the transcription factor BMAL1. Given the variety of potential targets for CKIepsilon and other cellular kinases, the precise role of phosphorylation is likely to be a complex one. Biochemical analysis of these and other circadian regulatory proteins has proven to be a fruitful approach in determining how they function within the context of the molecular clockworks. PMID- 15817303 TI - Nucleocytoplasmic shuttling of clock proteins. AB - The mammalian circadian clock in the neurons of suprachiasmatic nuclei (SCN) in the brain and in cells of peripheral tissues is driven by a self-sustained molecular oscillator, which generates rhythmic gene expression with a periodicity of about 24?h (Reppert and Weaver, 2002). This molecular oscillator is composed of interacting positive and negative transcription/translation feedback loops in which the heterodimeric transcription activator CLOCK?BMAL1 promotes the transcription of E-box containing Cryptochrome (Cry1 and Cry2) and Period (Per1 and Per2) genes, as well as clock-controlled output genes. After being synthesized in the cytoplasm, CRY and PER proteins feedback in the nucleus to inhibit the transactivation mediated by positive regulators. The mPER2 protein acts at the interphase between positive and negative feedback loops by indirectly promoting the circadian transcription of the Bmal1 gene (through RevErbalpha) (Preitner et al., 2002; Shearman et al., 2000) and by interacting with mCRY proteins (Kume et al., 1999; Yagita et al., 2002) (for a detailed review, see Reppert and Weaver, 2002). In addition to cyclic transcription of clock genes, immunohistochemical studies on SCN neurons have revealed that mCRY1, mCRY2, mPER1, and mPER2 proteins undergo near synchronous circadian patterns of nuclear abundance (Field et al., 2000). The delay of approximately 6h between the peak in clock mRNA production and maximal levels of protein expression in the nucleus is believed to originate from posttranslational modification steps involving phosphorylation, ubiquitination, and proteosomal degradation. Thus, the timing of entry, as well as the residence time of core clock proteins into the nucleus, is a critical step in maintaining the correct pace of the circadian clock. Several clock proteins have been shown to contain nuclear export signal, sequences, on top of nuclear import signals, that facilitate their cellular trafficking (Chopin Delannoy et al., 2003; Miyazaki et al., 2001; Yagita et al., 2002). This type of dynamic intracellular movement not only regulates protein localization, but also often affects functions by determining interactive partners and protein turnover. Because most of the clock genes have been identified by genetic screening in Drosophila and by gene knockdown in mammals, the development of innovative cellular techniques is essential in learning the structure-function and regulation of the corresponding proteins. This article discusses approaches, limitations, and applicable protocols to study the regulation of cellular localization of mammalian clock proteins, with a particular focus on mammalian CRY1 and PER2 proteins. PMID- 15817304 TI - Techniques that revealed the network of the circadian clock of Drosophila. AB - The techniques are reviewed that revealed the neuronal network of the circadian clock in the brain of the fruit fly as well as the function and localization of peripheral oscillators. Three principal techniques helped characterize the circadian clock network of Drosophila consisting of pacemaker centers in the brain and oscillators in peripheral tissues: (1) Immunolabeling with antibodies raised against specific clock proteins detected the tissues and cells that express the clock proteins, revealed the subcellular localization of clock molecules, and illuminated their abundance at different time points during the day; (2) reporter genes unraveled the network of clock neurons and reported the circadian cycling of the clock genes in vivo; and (3) genetic manipulations of clock gene expression elucidated the function of specific clock genes and clock cells. These techniques and the results gained by them are reviewed briefly. PMID- 15817306 TI - Transplantation of mouse embryo fibroblasts: an approach to study the physiological pathways linking the suprachiasmatic nucleus and peripheral clocks. AB - One of the unresolved issues in the field of circadian biology is dissection of the communication pathways between central and peripheral oscillators. We have developed an experimental procedure in which an implant of mouse embryo fibroblasts of a specific genotype can be successfully grafted into a host animal of a different genotype. This methodology provides an excellent tool to study how peripheral clocks are entrained under various physiological settings and the contribution of individual signaling effectors in this process. PMID- 15817305 TI - The suprachiasmatic nucleus is a functionally heterogeneous timekeeping organ. AB - Ever since the locus of the brain clock in the suprachiasmatic nucleus (SCN) was first described, methods available have both enabled and encumbered our understanding of its nature at the level of the cell, the tissue, and the animal. A combination of in vitro and in vivo approaches has shown that the SCN is a complex heterogeneous neuronal network. The nucleus is composed of cells that are retinorecipient and reset by photic input; those that are reset by nonphotic inputs; slave oscillators that are rhythmic only in the presence of the retinohypothalamic tract; endogenously rhythmic cells, with diverse period, phase, and amplitude responses; and cells that do not oscillate, at least on some measures. Network aspects of SCN organization are currently being revealed, but mapping these properties onto cellular characteristics of electrical responses and patterns of gene expression are in early stages. While previous mathematical models focused on properties of uniform coupled oscillators, newer models of the SCN as a brain clock now incorporate oscillator and gated, nonoscillator elements. PMID- 15817307 TI - Mouse chimeras and their application to circadian biology. AB - Chimeric mice are versatile model systems for the study of mammalian circadian biology. In chimeras, genetically different cells are combined within single animals, making them useful for assessing how normal cells interact with genetically altered cells in intact biological systems. In particular, the primary circadian pacemaker in the suprachiasmatic nucleus is amenable to analysis using series of chimeras that incorporate cells carrying mutations in circadian genes. The study of chimeras carrying circadian mutations can contribute to a better understanding of the function of the altered genes and of the fundamental physiology of circadian timing. Chimera analysis is a valuable approach for studying network properties in complex, integrated biological systems like that which controls circadian behavior in mammals. PMID- 15817308 TI - Measuring circadian rhythms in olfaction using electroantennograms. AB - Circadian clocks control daily rhythms in many behavioral, physiological, and metabolic processes. Despite remarkable advances in our understanding of the circadian timekeeping mechanism and how it responds to environmental cycles, relatively little is known about how the timekeeping mechanism regulates behavior, physiology, and metabolism. One of the most extensively characterized timekeeping mechanisms is that of Drosophila melanogaster. In this species, autonomous circadian clocks are found in many neuronal and nonneuronal tissues, including essentially all sensory structures. We have shown that sensory neurons in the antenna mediate a robust rhythm in electrophysiological responses to the food odorant ethyl acetate. This article describes how rhythms in olfactory responses are measured and provides a perspective on the generality of these rhythms and their regulation by the clock. PMID- 15817309 TI - Circadian effects of timed meals (and other rewards). AB - Mammals organize many of their activities around rhythmic events in their environments. Primary among these events is the daily light-dark cycle. However, for many animals, food availability is rhythmic or quasi-rhythmic and is therefore a potential synchronizing cue. While circadian rhythms in both behavior and physiological activity can be entrained in animals via meal-feeding schedules, the mechanism by which this occurs remains poorly understood. Similarities between the circadian effects of restricted feeding and the effects of chronic methamphetamine treatment may be indicative of a common mechanism. This article argues that reward (or the arousal that accompanies it) may be the final common pathway for such nonphotic circadian inputs. PMID- 15817310 TI - Peripheral clocks and the regulation of cardiovascular and metabolic function. AB - Circadian rhythms generated by cell autonomous biological clocks allow for the appropriate temporal synchronization of physiology and behavior, optimizing the efficiency of biological systems. Circadian oscillators and functions have been uncovered in both central and peripheral tissues. This article describes methodology, experimental design, and technical challenges pertaining to studies of circadian rhythms in the periphery. Experimental approaches are focused upon revealing the role of peripheral clocks in cardiovascular and metabolic function using in vitro and in vivo techniques. PMID- 15817311 TI - Circadian gene expression in cultured cells. AB - In mammals, circadian oscillators not only exist in specialized neurons of the suprachiasmatic nucleus, but in almost all peripheral cell types. These oscillators are operative even in established fibroblast cell lines, such as Rat 1 cells or NIH3T3 cells, and in primary fibroblasts from mouse embryos or adult animals. This can be demonstrated by treating such cells for a short time period with high concentrations of serum or chemicals that activate a large number of known signaling pathways. The possibility of studying circadian rhythms in cultured cells should facilitate the biochemical and genetic dissection of the circadian clockwork and should promote the discovery of new clock components. PMID- 15817312 TI - Cell culture models for oscillator and pacemaker function: recipes for dishes with circadian clocks? AB - Primary cell cultures of avian pinealocytes and the mammalian suprachiasmatic nucleus (SCN), immortalized cell lines derived from the SCN (SCN2.2), and fibroblasts derived from mice and rats have been employed as in vitro models to study the cellular and molecular mechanisms underlying circadian biological clocks. This article compares and contrasts these model systems and describes methods for avian pinealocyte cultures, immortalized SCN2.2 cells, and mouse fibroblast culture. Each of these culture models has advantages and disadvantages. Avian pinealocytes are photoreceptive, contain a circadian pacemaker, and produce rhythms of an easily assayed endocrine output-melatonin. However, the molecular mechanisms underlying pinealocyte function are not understood. SCN2.2 cells express metabolic and molecular rhythms and can impose rhythmicity on cocultured cells as well as rat behavior when transplanted into the brain. Yet, the entrainment pathways are not experimentally established in these cells. Fibroblast cultures are simple to produce and express molecular clock gene rhythms, but they express neither physiological rhythmicity nor pacemaker properties. The relative merits of these culture systems, as well as their impact on understanding circadian organization in vivo, are also considered. PMID- 15817313 TI - Analysis of circadian mechanisms in the suprachiasmatic nucleus by transgenesis and biolistic transfection. AB - Analysis of the cellular and molecular mechanisms that underlie the circadian pacemaker of the suprachiasmatic nuclei (SCN) requires in vitro preparations amenable to genetic manipulation that can provide dynamic measures of circadian activity in real time over multiple circadian cycles. This article focuses on the value of the SCN organotypic slice for such studies. Specifically, it describes the use of tissues from genetically modified mice in which the circadian promoter of the mPer1 gene is used to drive the expression of either firefly luciferase or destabilized green fluorescent protein optical reporters. Furthermore, we describe a procedure for biolistic (particle-mediated) transfection of SCN organotypic slices with fluorescent reporters that can be used to explore the cis acting elements and trans-acting factors that control circadian patterning, and also the interactions between subpopulations of neuronal oscillators within the SCN assemblage. PMID- 15817314 TI - Oligodeoxynucleotide methods for analyzing the circadian clock in the suprachiasmatic nucleus. AB - The recent identification of specific genes responsible for the generation of endogenous circadian rhythmicity in the suprachiasmatic nucleus presents a new level of investigation into endogenous rhythmicity and mechanisms of synchronization of this circadian clock with the environmental light?dark cycle. This article describes techniques that employ antisense and decoy oligodeoxynucleotides (ODN) to determine the roles of specific molecular substrates both in endogenous rhythmicity and in regulating the effects of light on the mammalian circadian clock. Application of antisense ODN technology has revealed a role for timeless (Tim) in the core clock mechanism and established that induction of period1 (Per1) is required for light responsiveness. Likewise, a decoy ODN designed to sequester activated CREB protein definitively demonstrated a requirement for CRE-mediated transcription in light signaling. Experiments designed with these molecular tools offer new insights on the interaction of cellular processes and signaling with the molecular clockworks. PMID- 15817315 TI - Assaying the Drosophila negative feedback loop with RNA interference in S2 cells. AB - Transcriptional negative feedback loops play a critical role in the molecular oscillations of circadian genes and contribute to robust behavioral rhythms. In one key Drosophila loop, CLOCK and CYCLE (CLK/CYC) positively regulate transcription of period (per). The period protein (PER) then represses this transcriptional activation, giving rise to the molecular oscillations of per RNA and protein. There is evidence that links molecular oscillations with behavioral rhythms, suggesting that PER also regulates the expression of downstream genes, ultimately resulting in proper behavior rhythmicity. Phosphorylation of PER has also been shown to be critical for rhythms. Doubletime (DBT) and casein kinase II (CKII) have been implicated in the phosphorylation of PER, which affects its stability as well as nuclear localization. We investigated the role of these kinases on PER transcriptional repression using the Drosophila S2 cell line in combination with RNA interference (RNAi) to knock down specific gene expression. This article describes the methods used to study PER repression activity in the S2 cell system as well as to exploit RNAi in this system. We also include protocols for immunocytochemistry and the application of leptomycin to differentiate direct effects on repression from indirect effects on subcellular localization. Finally, we discuss the generation of stable cell lines in the S2 cell system; these will be useful for experiments requiring homogeneous cell populations. PMID- 15817316 TI - Role of neuronal membrane events in circadian rhythm generation. AB - Circadian clock systems are composed of an input or "entrainment" pathway by which synchronization to the external environment occurs, a pacemaker responsible for generating rhythmicity, and an output or "expression" pathway through which rhythmic signals act to modulate physiology and behavior. The circadian pacemaker contains molecular feedback loops of rhythmically expressed genes and their protein products, which, through interactions, generate a circa 24-h cycle of transcription and translation of clock and clock-controlled genes. Neuronal membrane events appear to play major roles in entrainment of circadian rhythms in mollusks and mammals. In mammals, the suprachiasmatic nuclei of the hypothalamus receive photic information via the retinohypothalamic tract. Retinal signals, mediated by glutamate, induce calcium release and activate a number of intracellular cascades involved in photic gating and phase shifting. Membrane events are also involved in rhythm expression. Calcium and potassium currents influence the electrical output of pacemaker neurons by altering shape and intervals of impulse prepotentials, afterhyperpolarization periods, and interspike intervals, as well as altering membrane potentials and thereby shaping the spontaneous rhythmic spiking patterns. Unlike the involvement of membrane events in circadian entrainment and expression, it is less clear whether electrical activity, postsynaptic events, and transmembrane ion fluxes also are essential elements in rhythm generation. Studies, however, suggest that neuronal membrane activity may indeed play a crucial role in circadian rhythm generation. PMID- 15817317 TI - A screen for secreted factors of the suprachiasmatic nucleus. AB - The circadian clock in the suprachiasmatic nucleus (SCN) drives daily locomotor activity rhythms presumably by secreting diffusible factors whose target sites are accessible from the third ventricle of the hypothalamus. This article describes the methodology of a systematic molecular and behavioral screen to identify "locomotor factors" of the SCN. To find SCN-secreted factors not previously documented, a hamster SCN cDNA library was screened in a yeast signal sequence trap. In a subsequent behavioral screen, newly identified and previously documented SCN factors were tested for an effect on locomotor activity rhythms by chronic infusion into the third ventricle of hamsters. Using this approach combined with further experiments, we identified transforming growth factor-alpha (TGF-alpha) as a likely SCN inhibitor of locomotion. PMID- 15817318 TI - Genetic and biochemical strategies for identifying Drosophila genes that function in circadian control. AB - Explicit biochemical models have been elaborated for the circadian oscillators of cyanobacterial, fungal, insect, and mammalian species. In contrast, much remains to be learned about how such circadian oscillators regulate rhythmic physiological processes. This article summarizes contemporary genetic and biochemical strategies that are useful for identifying gene products that have a role in circadian control. PMID- 15817319 TI - Membranes, ions, and clocks: testing the Njus-Sulzman-Hastings model of the circadian oscillator. AB - Current circadian clock models based on interlocking autoregulatory transcriptional?translational negative feedback loops have arisen out of an explosion of molecular genetic data obtained over the last decade (for review, see Stanewsky, 2003; Young and Kay, 2001). An earlier model of circadian oscillation was based on feedback interactions between membrane ion transport systems and ion concentration gradients (Njus et al., 1974, 1976). This membrane model was posited as a more plausible alternative at the time to the even earlier "chronon" model, which was based on autoregulatory genetic feedback loops (Ehret and Trucco, 1967). The membrane model has been tested in a number of experimental systems by pharmacologically manipulating either ionic gradients across the plasma membrane or ion transport systems, but with inconsistent results. In the meantime, the scope and explanatory power of the genetic models overshadowed inquiries into the role of membrane ion fluxes in clock function. However, several recently developed techniques described in this article have provided a new glimpse into the essential role that membrane ion fluxes play in the mechanism of the core circadian oscillator and indicate that a complete understanding of the clock must include both genetic and membrane-based feedback loops. PMID- 15817320 TI - Mammalian photoentrainment: results, methods, and approaches. AB - Research on circadian biology over the past decade has paid increasing attention to the photoreceptor mechanisms that align the molecular clock to the 24-h light/dark cycle, and some of the results to emerge are surprising. For example, the rods and cones within the mammalian eye are not required for entrainment. A population of directly light-sensitive ganglion cells exists within the retina and acts as brightness detectors. This article provides a brief history of the discovery of these novel ocular photoreceptors and then describes the methods that have been used to study the photopigments mediating these responses to light. Photopigment characterization has traditionally been based on a number of complementary approaches, but one of the most useful techniques has been action spectroscopy. A photopigment has a discrete absorbance spectrum, which describes the probability of photons being absorbed as a function of wavelength, and the magnitude of any light-dependent response depends on the number of photons absorbed by the photopigment. Thus, a description of the spectral sensitivity profile (action spectrum) of any light-dependent response must, by necessity, match absorbance spectra of the photopigment mediating the response. We provide a step-by-step approach to conducting action spectra, including the construction of irradiance response curves, the calculation of relative spectral sensitivities, and photopigment template fitting, and discuss the underlying assumptions behind this approach. We then illustrate action spectrum methodologies by an in-depth analysis of action spectra obtained from rodless/coneless (rd/rd cl) mice and discuss, for the first time, the full implications of these findings. PMID- 15817321 TI - Cryptochromes and circadian photoreception in animals. AB - Cryptochromes are flavin- and folate-containing blue-light photoreceptors with a high degree of similarity to DNA photolyase, which repairs ultraviolet-induced DNA damage using blue light to initiate the repair reaction. Cryptochromes play essential roles in the maintenance of circadian rhythms in mice and Drosophila, and genetic data indicate that cryptochromes function as circadian photoreceptors in these and other animals. However, the photochemical reactions carried out by cryptochromes are not known at present. PMID- 15817322 TI - Nonvisual ocular photoreception in the mammal. AB - Rodents blind from outer retinal (rod and cone) degeneration still retain several light-dependent phenomena, including entrainment of the circadian clock and pupillary light responsiveness. This paradox is explained by the presence of intrinsically photosensitive retinal ganglion cells in the inner retina. These cells have unique properties, including a novel action spectrum, resistance to bleaching and adaptation under continuous light, and resistance to vitamin A depletion. Two candidate classes of photopigment have been proposed: melanopsin and cryptochromes. Physiologic analysis of circadian entrainment and pupillary light responsiveness in mice lacking these proteins leads to three conclusions: (1) outer and inner retinal photoreceptors provide partially redundant information to the inner retina, (2) melanopsin is required for inner retinal phototransduction in the absence of rod and cone signaling, and (3) cryptochromes contribute to the amplitude of inner retinal phototransduction but are not strictly required. PMID- 15817323 TI - Essentials of sleep recordings in Drosophila: moving beyond sleep time. AB - The power of Drosophila genetics can be used to facilitate the molecular dissection of sleep regulatory mechanisms. While evaluating total sleep time and homeostatic processes provides valuable information, other variables, such as sleep latency, sleep bout duration, sleep cycle length, and the time of day when the longest sleep bout is initiated, should also be used to explore the nature of a genetic lesion on sleep regulatory processes. Each of these variables requires that the recording interval used to identify periods of sleep and waking be determined accurately and empirically. This article describes the procedures for recording sleep in Drosophila and associated methodological constraints. In addition, it provides results from a normative data set of 1037 Canton-S female flies and 639 male flies to illustrate the nature and variability of sleep variables that one can extract from 24 h of data collection in Drosophila. PMID- 15817324 TI - Drosophila melanogaster: an insect model for fundamental studies of sleep. AB - In 2000, Drosophila melanogaster joined the ranks of vertebrates and invertebrates with a defined behavioral sleep state. The characterization of this sleep state revealed striking similarities to sleep in humans: sleep in flies has both circadian and homeostatic components, it is influenced by sex and age, and it is affected by pharmacological agents such as caffeine and antihistamines. As in mammals, arousal thresholds in flies increase with sleep deprivation. Furthermore, changes in brain electrical activity accompany the change from wake to sleep states. Not only do flies and vertebrates share these behavioral and physiological traits of sleep, but they are likely to share at least some genetic mechanisms underlying the regulation of sleep as well. This article reviews the methods currently used to identify and characterize the Drosophila sleep state. As these methods become more refined and our understanding of Drosophila sleep more detailed, the powerful techniques afforded by this organism are likely to unveil deep insights into the function(s) and regulatory mechanisms of sleep. PMID- 15817325 TI - Molecular evolution and population genetics of circadian clock genes. AB - This article discusses a number of common methodologies used in the field of population genetics and evolution and reviews their application within circadian rhythm research. We examine the basic principles behind phylogenetic analysis and how these can be used to illuminate clock gene evolution. We then discuss genetic variation between and within species and show how neutrality tests can reveal the signatures of selection or drift on clock genes. These tests are particularly important for moving beyond "just so" stories when discussing the evolution of clock phenotypes, and we provide relevant circadian examples. We also focus on methods that can be used to study genetic variation, such as quantitative trait loci analysis. We discuss the various bootstrapping or resampling techniques that can be applied to generate confidence intervals in the various methodologies and then examine the use of interspecific transformation studies, which can, and have, provide some useful insights, not only into clock gene evolution in particular, but "behavioral" gene evolution in general. Finally, we assess gene/protein alignments and protein structure predictions and their implicit evolutionary bases. PMID- 15817326 TI - Testing the adaptive value of circadian systems. AB - Circadian clocks are thought to enhance reproductive fitness. However, most of the evidence that supports the adaptiveness of clocks is not rigorous and falls into the category of "adaptive storytelling." Approaches that an evolutionary biologist would consider appropriate to address this issue are described along with an analysis of the evidence-past and present-that has been evoked to demonstrate the adaptive value of circadian systems. PMID- 15817327 TI - A "bottom-counting" video system for measuring cocaine-induced behaviors in Drosophila. AB - Cocaine exposure elicits a set of stereotypic behaviors in Drosophila that are strikingly similar to the cocaine-induced behaviors observed in vertebrates. This provides a valuable model for the study of cocaine abuse and has led to the discovery of a connection between the cocaine response pathway and the circadian system. This article describes a simplified assessment of cocaine-induced behavior combined with an image acquisition system, which allows the assay to be semiautomated. With this new system, cocaine response can be evaluated in a fraction of the time required by the previous assay, and subjectivity in scoring is reduced dramatically. PMID- 15817328 TI - The circadian clock and tumor suppression by mammalian period genes. AB - Period (Per) genes are key circadian rhythm regulators in mammals. Expression of mouse Per (mPer) genes has a diurnal pattern in the suprachiasmatic nucleus and in peripheral tissues. Genetic ablation mPER1 and mPER2 function results in a complete loss of circadian rhythm control based on wheel-running activity in mice. In addition, these animals also display apparent premature aging and a significant increase in neoplastic and hyperplastic phenotypes. When challenged by gamma radiation, mPer2-deficient mice respond by rapid hair graying, are deficient in p53-mediated apoptosis in thymocytes, and have robust tumor occurrences. Studies have demonstrated that the circadian clock function is very important for cell cycle, DNA damage response, and tumor suppression in vivo. The temporal expression of genes involved in cell cycle regulation and tumor suppression, such as c-Myc, Cyclin D1, Cyclin A, Mdm-2, and Gadd45alpha, is deregulated in mPer2 mutant mice. Genetic studies have demonstrated that many key regulators of cell cycle and growth control are also important circadian clock regulators, confirming the critical role of circadian function in organismal homeostasis. PMID- 15817329 TI - Androgen suppression adjuvant to definitive radiotherapy in prostate carcinoma- long-term results of phase III RTOG 85-31. AB - PURPOSE: Radiation Therapy Oncology Group protocol 85-31 was designed to evaluate the effectiveness of adjuvant androgen suppression, using goserelin, in unfavorable prognosis carcinoma of the prostate treated with definitive radiotherapy (RT). METHODS AND MATERIALS: Eligible patients were those with palpable primary tumor extending beyond the prostate (clinical Stage T3) or those with regional lymphatic involvement. Patients who had undergone prostatectomy were eligible if penetration through the prostatic capsule to the margin of resection and/or seminal vesicle involvement was documented histologically. Stratification was based on histologic differentiation, nodal status, acid phosphatase status, and prior prostatectomy. The patients were randomized to either RT and adjuvant goserelin (Arm I) or RT alone followed by observation and application of goserelin at relapse (Arm II). In Arm I, the drug was to be started during the last week of RT and was to be continued indefinitely or until signs of progression. RESULTS: Between 1987 and 1992, when the study was closed, 977 patients were entered: 488 to Arm I and 489 to Arm II. As of July 2003, the median follow-up for all patients was 7.6 years and for living patients was 11 years. At 10 years, the absolute survival rate was significantly greater for the adjuvant arm than for the control arm: 49% vs. 39%, respectively (p = 0.002). The 10-year local failure rate for the adjuvant arm was 23% vs. 38% for the control arm (p <0.0001). The corresponding 10-year rates for the incidence of distant metastases and disease-specific mortality was 24% vs. 39% (p <0.001) and 16% vs. 22% (p = 0.0052), respectively, both in favor of the adjuvant arm. CONCLUSION: In a population of patients with unfavorable prognosis carcinoma of the prostate, androgen suppression applied as an adjuvant after definitive RT was associated not only with a reduction in disease progression but in a statistically significant improvement in absolute survival. The improvement in survival appeared preferentially in patients with a Gleason score of 7-10. PMID- 15817330 TI - Biochemical failure and the temporal kinetics of prostate-specific antigen after radiation therapy with androgen deprivation. AB - PURPOSE: The accuracy of the American Society of Therapeutic Radiation Oncology consensus definition of biochemical failure (BF) after radiation therapy (RT) and androgen deprivation (AD) has been questioned, because posttreatment prostate specific antigen (PSA) levels typically rise after release from AD, and misclassification of BF may be made. The temporal kinetics of posttreatment PSA levels was examined to define the error in the classification of BF. METHODS AND MATERIALS: Between December 1, 1991 and April 30, 1998, 688 men with T1c-T3 NX/0 M0 prostate cancer received three-dimensional conformal RT alone (n = 586) or in combination with either short-term (STAD: 3 to 12 months, n = 82) or long-term (LTAD: 12 to 36 months, n = 20) AD. Follow-up, calculated from the end of all treatment, was >/=48 months. The mean posttreatment PSA was calculated in 3-month intervals. RESULTS: The median posttreatment clinical follow-up period was 76 months (range, 48-152 months). The posttreatment PSA values from the end of all treatment for the RT+STAD-BF group showed an initial period of rise followed by a period of decline at 30 months and then a continued rise again. The decline in the mean posttreatment PSA is explained in part by stabilization in PSA level after 3 consecutive rises. Nonbiochemical failures (NBF) after RT+STAD had a relatively constant mean PSA over time of approximately 0.5 ng/mL. Unlike the RT+STAD-NBF profile, the RT+LTAD-NBF profile rose continuously and steadily to a level approaching 1 ng/mL. The RT+LTAD-BF profile rose continuously but at a slower rate over time. Nine RT+STAD-NBF patients (22%) and 2 RT+LTAD-BF (29%) patients experienced 3 consecutive rises followed by a subsequent decline and stabilization of PSA compared to 10 RT-BF patients (5%). Redistributing these misclassified patients to their respective NBF groups changed the mean posttreatment PSA profiles as follows: The RT+LTAD-BF profile rose constantly and steadily with a doubling time of approximately 16 months, and the RT+LAD-NF initially rose to a value of approximately 0.5 ng/mL, then at 36 months began to decline. CONCLUSIONS: The temporal kinetics of posttreatment PSA after RT+AD and RT alone are different. The American Society of Therapeutic Radiation Oncology definition for biochemical failure overestimates BF in 20-30% after RT+AD compared to 5% after RT alone. PMID- 15817331 TI - Impact of short course hormonal therapy on overall and cancer specific survival after permanent prostate brachytherapy. AB - PURPOSE: To review the impact of prior hormonal therapy on 10-year overall and prostate cancer specific survival after primary brachytherapy. METHODS AND MATERIALS: A retrospective review was performed on the Arizona Oncology Services tumor registry for 2,378 consecutive permanent prostate brachytherapy cases from 1988 through 2001. Hormonal therapy was administered before the implant in 464 patients for downsizing of the prostate or at the discretion of the referring physician. All deceased patients with known clinical recurrence were considered to have died of prostate cancer, irrespective of the immediate cause of death. Risk groups were defined, with 1,135 favorable (prostate-specific antigen [PSA] < 10, Gleason < 7, Stage T1-T2a), 787 intermediate (single adverse feature), and 456 unfavorable (two or more adverse features) patients. Kaplan-Meier actuarial survival curves were generated for both overall and cause-specific survival from the time of treatment. Multivariate analysis was performed to assess the impact of hormonal intervention in comparison with known risk factors of grade, PSA, and age. RESULTS: With follow-up ranging up to 12.6 years and a median of 4.1 year, a total of 474 patients died, with 67 recorded as due to prostate cancer. Overall and cause-specific 10-year survival rates are 43% and 88%, respectively. Overall survival is 44% for the hormone naive patients, compared with 20% for the hormone treated cohort (p = 0.02). The cancer-specific survival is 89% vs. 81% for the same groups (p = 0.133). Multivariate analysis confirms the significance of age > 70 years (p = 0.0013), Gleason score >/= 7 (p = 0.0005), and prior hormone use (p = 0.0065) on overall survival. CONCLUSIONS: At 10 years, in prostate cancer patients receiving brachytherapy, overall survival is worse in men receiving neoadjuvant hormonal therapy, compared with hormone naive patients. This does not appear to be due to other known risk factors for survival (i.e., stage, grade, PSA, age) on multivariate analysis. The leading causes of death were cardiovascular, prostate cancer, and other cancers with no obvious discrepancy between the two groups. This finding is unexpected and requires confirmation from other centers. PMID- 15817332 TI - High-dose-rate intensity-modulated brachytherapy with external beam radiotherapy for prostate cancer: California endocurietherapy's 10-year results. AB - PURPOSE: To present the long-term outcome and morbidity of high-dose-rate brachytherapy (HDR-BT) combined with external beam radiotherapy (EBRT) for localized prostate cancer. METHODS AND MATERIALS: Between September 1991 and December 1998, 209 consecutive patients with no prior androgen suppression were treated with HDR-BT plus EBRT. The median follow-up was 7.25 years (range, 5-12 years). The patients were stratified into three risk groups: low (Stage T2a or less, Gleason score 20). Four definitions of PSA progression were compared with the general clinical failure outcome: the American Society for Therapeutic Radiology and Oncology (ASTRO) definition, nadir plus 2.0 ng/mL, two consecutive rises >/=0.5 ng/mL, and PSA level >0.2 ng/mL. Morbidity was scored using Radiation Therapy Oncology Group criteria. RESULTS: The general clinical control rate was 90% (188 of 209), and the general clinical failure rate was 10% (21 of 209). The overall survival rate was 79%, and the cause-specific survival rate was 97%. The PSA progression-free survival (ASTRO definition) rate was 90%, 87%, and 69% for the low-, intermediate-, and high-risk groups, respectively. The nadir plus 2 ng/mL and two rises >/=0.5 definitions correlated better with the actual clinical outcome than did the ASTRO and PSA >0.2 ng/mL definitions. The rate of Grade 3 and 4 late urinary morbidity was 6.7% and 1%, respectively, mostly occurring in patients who had undergone post-RT transurethral prostate resection. No late Grade 3 or 4 rectal morbidity developed. The sexual potency preservation rate was 67%. CONCLUSION: Our 10-year results have demonstrated HDR-BT plus EBRT is a proven treatment for all stages of localized prostate cancer. The morbidity was low, but post-RT transurethral resection should be avoided. PMID- 15817333 TI - Conservative surgery and radiotherapy for stage I/II breast cancer using lung density correction: 10-year and 15-year results. AB - PURPOSE: Radiotherapy (RT) planning for breast cancer using lung density correction improves dose homogeneity. Its use obviates the need for a medial wedge, thus reducing scatter to the opposite breast. Although lung density correction is used at many centers in planning for early-stage breast cancer, long-term results of local control and survival have not been reported. Since 1984, we have used lung density correction for dose calculations at the University of Michigan. We now present our 10-year and 15-year results. METHODS AND MATERIALS: The records of 867 patients with Stage I/II breast cancer treated with breast-conserving surgery and RT with or without systemic therapy were reviewed. Tangential fields delivering 45-50 Gy to the whole breast calculated using lung density correction were used. A boost was added in 96.8% of patients for a total median dose of 61.8 Gy. RESULTS: With a median follow-up of 6.6 years (range, 0.2-18.9 years), 5-, 10-, and 15-year actuarial rates of in-breast tumor recurrence as only first failure were 2.2%, 3.6%, and 5.4%, respectively. With surgical salvage, the 15-year cumulative rate of local control was 99.7%. Factors that significantly predicted for increased rate of local recurrence in multivariate analysis were age 20% was identified in women with one to three positive nodes plus at least one of the following factors: age <45 years, Stage T2, histologic Grade 3, ER negative disease, medial location, more than one positive node, or >25% of nodes positive (all p < 0.05 on univariate analysis). On multivariate analysis, age <45 years, >25% of nodes positive, medial tumor location, and ER-negative status were statistically significant predictors of isolated LRR and LRR +/- SDR. In the classification model, the first split was according to age (<45 years vs. >/=45 years), with 29.3% vs. 13.7% developing LRR +/- SDR (p < 0.0001). Of 123 women <45 years, the presence of >25% of nodes positive was associated with a risk of LRR +/- SDR of 58.0% compared with 23.8% for those with 45 years, the presence of >25% of nodes positive also conferred a greater LRR +/- SDR risk (26.7%) compared with women with 45 years with 25% of nodes positive, a medial tumor location, and ER-negative status were statistically significant independent factors associated with greater LRR, meriting consideration and discussion of PMRT. Combinations of these factors further augmented the LRR risk, warranting recommendation of PMRT to optimize locoregional control and potentially improve survival. The absence of high-risk factors identifies women who may reasonably be spared the morbidity of PMRT. PMID- 15817336 TI - Breast radiotherapy in the lateral decubitus position: A technique to prevent lung and heart irradiation. AB - PURPOSE: To present an original technique for breast radiotherapy, with the aim of limiting lung and heart irradiation, satisfying quality assurance criteria. METHODS AND MATERIAL: An original radiotherapy technique for breast irradiation has been developed at the Institute Curie in January 1996. It consists of isocentric breast irradiation in the lateral decubitus position (isocentric lateral decubitus [ILD]). This technique is indicated for voluminous or pendulous breasts needing breast irradiation only. Thin carbon fiber supports and special patient positioning devices have been developed especially for this technique. In vivo measurements were performed to check the dose distribution before the routine use of the technique. RESULTS: ILD has been successfully implemented in routine practice, and 500 patients have been already treated. Breast radiotherapy is performed using a dose of 50 Gy at ICRU point in 25 fractions. ILD shows good homogeneity of the dose in breast treatment volume, treatment fields are perpendicular to the skin ensuring its protection, and extremely low dose is delivered to the underlying lung and heart. CONCLUSION: In cases of voluminous breasts or patients with a history of lung and heart disease, our technique provides several advantages over the conventional technique with opposing tangential fields. This technique improves the dose homogeneity according to the ICRU recommendations. PMID- 15817337 TI - Dose-volume histogram analysis as predictor of radiation pneumonitis in primary lung cancer patients treated with radiotherapy. AB - PURPOSE: To determine the relationship between various parameters derived from lung dose-volume histogram analysis and the risk of symptomatic radiation pneumonitis (RP) in patients undergoing radical radiotherapy for primary lung cancer. METHODS AND MATERIALS: The records of 156 patients with lung cancer who had been treated with radical radiotherapy (>/=45 Gy) and for whom dose-volume histogram data were available were reviewed. The incidence of symptomatic RP was correlated with a variety of parameters derived from the dose-volume histogram data, including the volume of lung receiving 10 Gy (V(10)) through 50 Gy (V(50)) and the mean lung dose (MLD). RESULTS: The rate of RP at 6 months was 15% (95% confidence interval 9-22%). On univariate analysis, only V(30) (p = 0.036) and MLD (p = 0.043) were statistically significantly related to RP. V(30) correlated highly positively with MLD (r = 0.96, p < 0.001). CONCLUSION: V(30) and MLD can be used to predict the risk of RP in lung cancer patients undergoing radical radiotherapy. PMID- 15817338 TI - Phase I trial of Orzel (UFT plus leucovorin), cisplatin, and radiotherapy in the treatment of potentially resectable esophageal cancer. AB - PURPOSE: Fluorinated pyrimidines have been established as radiosensitizers in the combined modality therapy of esophageal cancer. UFT, an oral combination of a 5 fluorouracil pro-drug (uracil) and a dihydropyrimidine dehydrogenase inhibitor (ftorafur), may provide improvement in the ease of administration with equal efficacy. This Phase I study was designed to determine the maximal tolerated dose and dose-limiting toxicity of UFT, leucovorin, and cisplatin when given with radiotherapy in the neoadjuvant treatment of resectable esophageal cancer. METHODS: Chemotherapy consisted of i.v. cisplatin 80 mg/m(2) (Days 1 and 22) and UFT with leucovorin orally on Days 1-35. UFT was escalated in 50-mg/m(2) increments, starting at 200 mg/m(2)/d. Radiotherapy consisted of 4500 cGy in 25 fractions. Patients underwent resection 4-6 weeks after chemoradiotherapy. RESULTS: Ten patients with resectable esophageal cancer were enrolled. Of the 7 patients entered at dose level 1, 1 developed a dose-limiting toxicity of nausea. All 3 patients entered at dose level 2 developed dose-limiting toxicity. The maximal tolerated dose for UFT was the starting level, 200 mg/m(2)/d. Of the 10 patients enrolled, 8 underwent esophagectomy and 2 developed progressive disease and did not undergo surgery. The disease of 6 of the 8 patients was downstaged at surgery. CONCLUSION: The recommended UFT dose for Phase II studies is 200 mg/m(2)/d given orally in two divided doses when given with leucovorin, cisplatin, and radiotherapy. PMID- 15817339 TI - Preoperative radiotherapy for rectal adenocarcinoma: Which are strong prognostic factors? AB - PURPOSE: This retrospective 12-year study evaluated the prognostic value of initial and postoperative staging of rectal tumors. METHODS AND MATERIALS: Between 1985 and 1996, 297 patients were treated with preoperative radiotherapy (39 Gy in 13 fractions) and surgery for Stage T2-T4N0-N1M0 rectal adenocarcinoma. Pretreatment staging included a clinical examination and endorectal ultrasonography (EUS) since 1988. Clinical staging was performed by digital rectal examination and rigid proctoscopy. EUS was performed in 236 patients. Postoperative staging was performed by examination of the pathologic specimen. RESULTS: The median follow-up was 49 months. The overall 5-year survival rate was 67%, with a local failure rate of 9%. The rate of sphincter preservation was 65%. The clinical examination findings were strong prognostic factor for both cT stage (p < 0.001) and cN stage (p < 0.006) but had poor specificity for cN stage (only 25 lymph nodes detected). In both univariate and multivariate analyses, EUS had a statistically significant prognostic value for uT (p < 0.014) but not for uN (p < 0.47) stage. In contrast, pT and pN stages were strong prognostic factors (p < 0.001 and p < 0.001, respectively). CONCLUSION: Pretreatment staging, including clinical examination and EUS, seemed accurate enough to present a high prognostic value for the T stage. EUS was insufficient to stage lymph node involvement. Owing to its lack of specificity, uN stage was not a reliable prognostic factor. An improvement in N staging is necessary and essential. Despite downstaging, postoperative staging remained a very strong prognostic factor for both T and N stages. PMID- 15817340 TI - Preoperative chemoradiation with oral tegafur within a multidisciplinary therapeutic approach in patients with T3-4 rectal cancer. AB - PURPOSE: The aim of this study was to evaluate the activity in terms of downstaging histologic patterns of residual tumor and clinical tolerance of a neoadjuvant chemoradiation program with oral tegafur for rectal cancer. METHODS AND MATERIALS: From May 1998 to May 2001, 62 consecutive patients with cT(3-4) or cN(+) rectal cancer, or both, were treated with 45-50 Gy (1.8 Gy/day; 25 fractions) and oral tegafur 1200 mg/day. Surgery was performed 6 weeks after the completion of chemoradiation. All patients received a boost with intraoperative electron beam radiotherapy (IOERT) over the presacral space. RESULTS: Grade 3-4 hematologic toxicity consisted on Grade 3 anemia in 1 patient. Nonhematologic toxicity was mild. Fifteen patients (23%) had Grade 3 dermatitis, 16 (25%) had Grade 3, and 2 (3%) had Grade 4 proctitis. The median dose of radiotherapy was 50.4 Gy. Surgery consisted on anterior resection in 38 patients (61%) and abdominoperineal amputation in 24 (39%). Five complete pathologic responses were observed (8%), and 29 patients (47%) had a minimal microscopic residual tumor (mic category). The total downstaging rate was 68%. With a median follow-up of 46 months, the pelvic control rate was 95%, disease-free survival 74.1%, and overall survival 76.5%. CONCLUSIONS: Neoadjuvant chemoradiation with oral tegafur is feasible, well tolerated, and active, with the additional advantage of offering the convenience of oral chemotherapy. PMID- 15817341 TI - Comparison of CT- and FDG-PET-defined gross tumor volume in intensity-modulated radiotherapy for head-and-neck cancer. AB - PURPOSE: To compare the gross tumor volume (GTV) identified on CT to that obtained from fluorodeoxyglucose (FDG) positron emission tomography (PET) and determine the differences in volume and dose coverage of the PET-GTV when the CT GTV is used for radiotherapy planning. METHODS AND MATERIALS: A total of 40 patients with intact squamous cell carcinoma arising in the head-and-neck region underwent intensity-modulated radiotherapy (IMRT) at one department. All patients underwent CT simulation for treatment planning followed by PET-CT in the treatment position. CT simulation images were fused to the CT component of the PET-CT images. The GTV using the CT simulation images was contoured (CT-GTV), as was the GTV based on the PET scan (PET-GTV). The IMRT plans were obtained using the CT-GTV. RESULTS: The PET-GTV was smaller, the same size, and larger than the CT-GTV in 30 (75%), 3 (8%), and 7 (18%) cases respectively. The median PET-GTV and CT-GTV volume was 20.3 cm(3) (range, 0.2-294) and 37.2 cm(3) (range, 2-456), respectively. The volume of PET-GTV receiving at least 95% of the prescribed dose was 100% in 20 (50%), 95-99% in 10 (25%), 90-94% in 3 (8%), 85-89% in 1 (3%), 80 84% in 2 (5%), 75-79% in 1 (3%), and <75% in 3 (8%) cases. The minimal dose received by 95% of the PET-GTV was >/=100% in 19 (48%), 95-99% in 11 (28%), 90 94% in 5 (13%), 85-89% in 2 (5%), and <75% in 3 (8%) cases. CONCLUSION: The PET GTV was larger than the CT-GTV in 18% of cases. In approximately 25% of patients with intact head-and-neck cancer treated using IMRT, the volume of PET-GTV receiving at least 95% of the prescribed dose and minimal dose received by 95% of the PET-GTV were less than optimal. PMID- 15817342 TI - Prospective study of inner ear radiation dose and hearing loss in head-and-neck cancer patients. AB - PURPOSE: To determine the relationship between the radiation dose to the inner ear and long-term hearing loss. METHODS AND MATERIALS: Eligible patients included those receiving curative radiotherapy (RT) for head-and-neck cancer. After enrollment, patients underwent three-dimensional conformal RT planning and delivery (180-200 cGy/fraction) appropriate for their disease site and stage. The inner ear was contoured on axial CT planning images. Dose-volume histograms, as well as the mean and maximal dose for each structure, were calculated. Patients underwent pure tone audiometry at baseline (before treatment) and 1, 6, 12, 24, and 36 months after RT. The threshold level (the greater the value, the more hearing loss) in decibels was recorded for 250, 500, 1000, 2000, 4000, and 8000 Hz. For patients receiving predominantly unilateral RT, the contralateral ear served as the de facto control. The differences in threshold level between the ipsilateral and contralateral ears were calculated, and the temporal pattern and dose-response relation of hearing loss were analyzed using statistical methods that take into account the correlation between two ears in the same subject and repeated, sequential measurements of each subject. RESULTS: Of the 40 patients enrolled in this study, 35 qualified for analysis. Four patients who received concurrent chemotherapy and RT were analyzed separately. The 31 unilaterally treated patients received a median dose of 47.4 Gy (range, 14.1-68.8 Gy) to the ipsilateral inner ear and 4.2 Gy (range, 0.5-31.3 Gy) to the contralateral inner ear. Hearing loss was associated with the radiation dose received by the inner ear (loss of 210dB was observed in ears receiving >/=45 Gy) and was most appreciable in the higher frequencies (>/=2000 Hz). For a 60-year-old patient with no previous hearing loss in either ear, after receiving 45 Gy, the ipsilateral ear, according to our clinical model, would have a 19.3-dB (95% confidence interval [CI], 15.5-23.0) and 5.4-dB (95% CI, 3.5-7.5) hearing decrement compared with the contralateral ear for 8000 Hz and 1000 Hz, respectively. Age and an initial hearing difference within an ear pair also affected hearing loss. The baseline hearing threshold was inversely related to radiation-induced hearing loss. The degree of hearing loss was dependent on the frequency tested, age, baseline hearing, and baseline difference in hearing between a patient's two ears. CONCLUSION: High-frequency (>/=2000 Hz) hearing acuity worsens significantly after RT in a dose-dependent fashion. A larger number of patients needs to be studied to validate these results. This knowledge can be applied to create guidelines regarding future dose limits to the auditory apparatus for patients undergoing head-and-neck RT. PMID- 15817343 TI - Postradiotherapy quality of life for head-and-neck cancer patients is independent of xerostomia. AB - PURPOSE: To determine the relationship between quality of life (QOL) and xerostomia over time for patients undergoing radiotherapy (RT) for head-and-neck cancer in a prospective clinical trial. METHODS AND MATERIALS: Patients with head and-neck cancer were randomized to pilocarpine (n = 65) vs. placebo (n = 65) during RT. QOL was measured using the McMaster Head and Neck Radiotherapy Questionnaire (HNRQ). Xerostomia was measured on a linear analog scale. No statistically significant differences were observed between arms; all 130 patients were analyzed together. RESULTS: Baseline QOL data were obtained for 98.5% of participants. The baseline HNRQ score of 5.7 declined significantly to 4.0 (p <0.0001) by RT Week 6 and returned to baseline (5.8) by 6 months after treatment. This represents a large, clinically important change of 1.7 of 7 (24%; effect size 1.34). The decline in HNRQ score during RT paralleled the onset of xerostomia on the linear analog scale (r = 0.36 at 1 month). After treatment, the QOL scores recovered without improvement in xerostomia. The trajectory of the linear analog scale score resembled that of the HNRQ's single xerostomia question (r = 0.75 at 1 month). CONCLUSION: Quality of life recovers to baseline after RT, despite persistent xerostomia. Either a response shift occurs or xerostomia in the absence of acute mucositis has a relatively small influence on overall QOL. PMID- 15817344 TI - Nasopharyngeal cancer in the Middle East: experience of the American University of Beirut Medical Center. AB - PURPOSE: To review the data of nasopharyngeal carcinoma (NPC) treated at the American University of Beirut Medical Center and reflect on the characteristics and treatment outcome of NPC in the Middle East compared with those of Western countries and countries in which NPC is endemic. METHODS AND MATERIALS: Between 1966 and 1998, 151 patients with the diagnosis of NPC received definitive radiotherapy at the American University of Beirut Medical Center. Of the 151 patients, 111 were males (gender ratio, 2.78); the median age was 45 years (range, 11-75 years). Most (95%) patients (n = 144) were Lebanese, 4 were Syrians, and 3 were from the Gulf countries. Most (60%) patients (n = 91) had Stage IV disease, 27% had Stage III, and 13% had Stage I or II disease; nodal disease was present in 117 patients (77%). The pathologic type was predominantly lymphoepithelioma or World Health Organization type III (95 patients, 63%). Treatment consisted of definitive radiotherapy alone for 116 patients (77%). All others received induction chemotherapy, primarily with cisplatin-containing regimens. The median radiation dose was 66 Gy (range, 47-73 Gy) to the primary and 67 Gy (range, 49-85 Gy) to involved neck nodes given at 2 Gy/fraction. The average follow-up was 3.02 years (range, 0.1-24.5 years). RESULTS: The 5-year and 10-year disease-free survival (DFS) rate was 46%. Using univariate analyses, the following factors significantly affected DFS: node size (<3 vs. 3-6 vs. >6 cm; p = 0.01), node level (upper vs. mid vs. lower neck; p = 0.004), and duration of radiotherapy (p = 0.002). However, T stage, age, gender, radiation dose, use of chemotherapy, and histologic features had no statistically significant influence on DFS. The actuarial rate of local control at 5 and 10 years was 81% and 73%. T stage, N stage, and histologic features were statistically significant variables for local control in the univariate analyses. Using a Cox regression model, N stage (N1-N2 vs. N3; relative risk 2.09, p = 0.004) was identified as an independent variable for DFS, and N stage and pathologic features were identified as independent variables for local control. The actuarial rate of distant metastases was 32% at both 5 and 10 years. Distant metastases were only affected by N stage (upper-mid vs. lower neck; p = 0.004). Six patients (4%) were reported to have Grade 4 late complications. CONCLUSION: Our results indicate that the characteristics of NPC patients in Lebanon and their parameters of outcome are comparable to those reported in Western series, particularly for the relative frequency and effect of lymphoepithelial histologic type. Because of potential confounding factors, no definite conclusions about induction chemotherapy could be drawn from this retrospective study. PMID- 15817345 TI - Results of treatment intensification for progressive locoregional disease in head and-neck cancer patients undergoing postoperative radiotherapy. AB - PURPOSE: Patients who develop progressive locoregional disease during radical surgery and postoperative radiotherapy for squamous cell carcinoma of the head and neck represent a management dilemma. We present our experience using treatment intensification for such patients. METHODS AND MATERIALS: A prospective record of eligible patients was kept between May 1998 and December 2001. The study included 15 patients, 11 men and 4 women (median age, 60 years); 67% had Stage III-IV disease. The sites of progression were primary in 3, nodes/scar in 10, and both primary and nodes in 2. Relative to the initial plan, treatment intensification was achieved by an increased radiation dose in 7 (using accelerated fractionation in 5), an increased radiation dose and the addition of concomitant chemotherapy in 7, and the addition of concomitant chemotherapy alone in 1 patient. RESULTS: The median follow-up was 40 months. Eight patients had a complete response to intensified treatment. At the closeout date, 6 patients were alive with no evidence of disease. Eight patients had died with locoregional disease; two also had distant metastases. One patient was lost to follow-up after achieving a complete response. The median failure-free survival for all patients was 6 months, but for those with a complete response, it was 37 months. The median overall survival time was 28 months. The 2-year and 3-year overall survival rate was 50% and 42%, respectively. Acute mucosal and skin toxicity was increased relative to standard postoperative radiotherapy but was not dissimilar to that expected after radical definitive chemoradiotherapy. CONCLUSION: Intensification of treatment in patients who develop progressive locoregional disease is warranted, because it can lead to long-term disease control in a subset of patients with significant but acceptable toxicity. PMID- 15817346 TI - Reirradiation alternating with docetaxel and cisplatin in inoperable recurrence of head-and-neck cancer: a prospective phase I/II trial. AB - PURPOSE: Inoperable locoregional recurrences of head-and-neck cancer in a previously irradiated volume represent a therapeutic dilemma. Chemotherapy alone has no curative potential, whereas reirradiation and concurrent chemoradiation can salvage a small fraction of patients. Mucosal toxicity of concurrent chemoradiation requires substantial dose reduction of chemotherapy. Alternating chemoradiation offers the chance to give both full-dose chemotherapy and radiotherapy. The latter may provide a particular advantage for recurrent, potentially radiation resistant tumors. The feasibility and efficacy of a full dose docetaxel containing alternating chemoradiation schedule was tested. PATIENTS AND METHODS: Twenty-seven patients (Karnofsky performance status score >/=70%) with histologically proven recurrent squamous cell cancer that occurred >/= 6 months in a previously irradiated area (>/= 60 Gy) were considered unresectable and unsuitable for brachytherapy. Alternating chemoradiation consisted of 3 cycles of docetaxel 60 mg/m(2) d1 and cisplatin 15 mg/m(2) d2-5, q d22, and involved field radiotherapy 2.0 Gy every day d8-12, d15-19, d29-33, and d36-40 (40.0 Gy total dose). Dose reduction of docetaxel to 50 mg/m(2) was necessary, because of hematologic toxicity in the first 12 patients. RESULTS: Alternating chemoreirradiation was applied as planned in 12 of 27 patients, with reirradiation completed per protocol in 81%. Overall, patients received 83% of the intended dose of docetaxel and 73% of cisplatin. Third-degree common toxicity criteria mucositis occurred in 15%, leukopenia of >/= third degree by common toxicity criteria in 37%, and 3 early deaths were observed. Median time to follow up, time to local progression, median survival, and 3-year survival rates were 42 months, 10 months, 10 months, and 18%, respectively. CONCLUSIONS: Alternating chemoreirradiation in recurrences of head-and-neck cancer resulted in 80% overall response with acceptable toxicity. A significant minority of patients had durable tumor control with a chance of long-term survival. PMID- 15817347 TI - Intensity-modulated radiotherapy for head-and-neck rhabdomyosarcoma. AB - PURPOSE: To determine the preliminary results of intensity-modulated radiotherapy (IMRT) for head-and-neck rhabdomyosarcoma. METHODS AND MATERIALS: Twenty-eight patients underwent IMRT as a part of multimodality therapy. Twenty-one tumors were parameningeal, three were orbital, and four were in other sites. The median age was 8 years (range, 1-29 years). Most (89%) had Group III disease. Intracranial extension was present in 71% of parameningeal tumors. A 1.5-cm margin was used, and the median dose was 50.4 Gy (range, 30-55.8 Gy). RESULTS: The actuarial 3-year survival rate for patients with parameningeal tumors was 65%. The 3-year actuarial freedom from failure rate was 95% locally, 90% in regional nodes, 88% in the central nervous system, and 80% at distant sites. No failures occurred among patients with orbit tumors; a single central nervous system failure occurred in 1 patient with a lip/cheek tumor. Disease-free survival was significantly worse for patients with alveolar histologic features (p = 0.01). Acute radiation toxicity was similar to that reported by the Intergroup Rhabdomyosarcoma Study Group. Late radiation toxicity was recorded and was mild. CONCLUSION: IMRT with image fusion results in outstanding local control despite the use of a reduced margin. However, survival among patients with alveolar histologic findings or intracranial extension remains unacceptably low. PMID- 15817348 TI - Histologic assessment of peritumoral edema in soft tissue sarcoma. AB - PURPOSE: To evaluate whether satellite tumor cells can be identified histologically in the tissues surrounding a soft tissue sarcoma and whether their presence correlates with increased T(2)-weighted signal intensity on MRI. METHODS AND MATERIALS: Fifteen patients with a high-grade extremity or truncal soft tissue sarcoma underwent preoperative MRI. The extent of high T(2)-weighted signal changes in the tissues surrounding tumor, thought to represent peritumoral edema/reactive changes, was determined. Twelve patients received i.v. gadolinium, and contrast enhancement was determined. All patients underwent surgical resection in the absence of preoperative chemotherapy or radiotherapy. The presence of tumor cells in the surrounding tissues was determined histologically in representative paraffin-embedded sections and correlated with the MRI findings. RESULTS: The extent of peritumoral T(2)-weighted MRI signal changes ranged from 0 to 7.1 cm (mean, 2.5 cm); contrast enhancement ranged from 0 to 5.3 cm (mean, 1.1 cm). Sarcoma cells were identified histologically in the tissues beyond the tumor in 10 of 15 cases. In 6 cases, tumor cells were located within 1 cm of the tumor margin, and in 4 cases, malignant cells were found at a distance >1 cm and up to a maximum of 4 cm. The location of tumor cells beyond the margin did not correlate with tumor size nor did it correlate with the location or extent of peritumoral changes. CONCLUSION: The ability to identify tumor cells beyond the margin of a soft tissue sarcoma has important implications in planning appropriate targets for treatment. This could influence the use of new radiotherapy technologies such as intensity-modulated radiotherapy that aim to minimize treatment volumes through conformal planning. PMID- 15817349 TI - Angiosarcoma of the scalp treated with curative radiotherapy plus recombinant interleukin-2 immunotherapy. AB - PURPOSE: To evaluate the effectiveness of curative radiotherapy (RT) plus recombinant interleukin-2 (rIL-2) immunotherapy regarding the treatment results for angiosarcoma of the scalp. Curative resection of angiosarcoma of the scalp is usually difficult because of the diffuse, clinically undetectable local spread. RT is a rational therapeutic approach, because a wide region of the dermis can be treated, while sparing the underlying normal tissues. Recently, the effectiveness of immunotherapy with rIL-2 has also been reported in the treatment of angiosarcoma of the scalp. METHODS AND MATERIALS: The data of 20 patients with angiosarcoma of the scalp treated with curative RT plus rIL-2 immunotherapy between January 1988 and June 2002 were retrospectively analyzed. The total radiation dose was 70.3 +/- 6.9 Gy. The fractions were 2-3 Gy daily, given 5 d/wk. rIL-2 immunotherapy was performed by transcatheter arterial administration in 10 patients, systemic administration in 11 during the course of RT, and intratumoral injection in 10 during and/or after RT; 12 patients received a combination of two. Five patients underwent limited surgery, and concomitant pacilitaxel chemotherapy was also used in 2 patients. RESULTS: The median survival time for overall, local recurrence-free, and distant metastasis-free survival was 36.2, 11.1, and 17.8 months, respectively. Local recurrence developed in 7 patients (35%), 4 of whom also had evidence of distant metastases. An additional 7 patients (35%) developed distant metastases alone. Recurrence within the radiation field was recognized in 2 patients with systemic rIL-2 administration alone (p < 0.05). Arterial or intratumoral administration combined with systemic administration of rIL-2 resulted in better distant metaststasis free survival rates (p < 0.05). CONCLUSION: Curative RT plus rIL-2 immunotherapy provided an efficient, effective means of treating angiosarcoma of the scalp. Arterial or intratumoral administration combined with systemic administration of rIL-2 may prolong survival. Additional studies with detailed treatment protocols are recommended. PMID- 15817350 TI - A phase II study of concurrent temozolomide and cis-retinoic acid with radiation for adult patients with newly diagnosed supratentorial glioblastoma. AB - PURPOSE: This Phase II study was designed to determine the median survival time of adults with supratentorial glioblastoma treated with a combination of temozolomide (TMZ) and 13-cis-retinoic acid (cRA) given daily with conventional radiation therapy (XRT). METHODS AND MATERIALS: This was a single arm, open labeled, Phase II study. Patients were treated with XRT in conjunction with cRA and TMZ. Both drugs were administered starting on Day 1 of XRT, and chemotherapy cycles continued after the completion of XRT to a maximum of 1 year. RESULTS: Sixty-one patients were enrolled in the study. Time to progression was known for 55 patients and 6 were censored. The estimated 6-month progression-free survival was 38% and the estimated 1-year progression-free survival was 15%. Median time to progression was estimated as 21 weeks. The estimated 1-year survival was 57%. The median survival was 57 weeks. CONCLUSIONS: The combined therapy was relatively well tolerated, but there was no survival advantage compared with historical studies using XRT either with adjuvant nitrosourea chemotherapy, with TMZ alone, or with the combination of TMZ and thalidomide. Based on this study, cRA does not seem to add a significant synergistic effect to TMZ and XRT. PMID- 15817351 TI - Hypofractionated conformal stereotactic radiotherapy alone or in combination with whole-brain radiotherapy in patients with cerebral metastases. AB - PURPOSE: The aim was to evaluate treatment of cerebral metastases with hypofractionated conformal stereotactic radiotherapy (HCSRT) or whole-brain radiotherapy (WBRT) in combination with a stereotactic boost. METHODS AND MATERIALS: Forty-seven patients were treated with HCSRT and 14 patients with WBRT in combination with a stereotactic boost. Radiation doses were 40 Gy (5 fractions) in HCSRT or 30 Gy (WBRT) combined with a mean dose of 17 Gy stereotactically (1-3 fractions). RESULTS: The median survival time in the HCSRT as well as the WBRT group was 5.0 months, and 87% died of extracranial disease. Radiologic follow-up (mean, 3.7 months after treatment) showed local control in the HCSRT group in 84% and in the WBRT group in 100%. Patients treated with HCSRT developed new brain metastases distant from the irradiated area in 25%. Two patients treated with HCSRT deteriorated neurologically during treatment, and in 2 patients radionecrosis developed. CONCLUSIONS: Although there may be a higher risk of distant new metastases, HCSRT as a treatment for brain metastases seems to be as effective as WBRT in combination with a stereotactic boost. Complications are in the range of what has been reported previously. PMID- 15817352 TI - Initial clinical experience with frameless radiosurgery for patients with intracranial metastases. AB - PURPOSE: To review the initial clinical experience with frameless stereotactic radiosurgery (SRS) for treating intracranial metastatic disease. METHODS AND MATERIALS: Sixty-four patients received frameless SRS for intracranial metastatic disease. Minimum follow-up was 6 months with none lost to follow-up. Patients had a median of 2 metastases and a maximum of 4. The median number of isocenters was 2 with median arcs of 10 and median dose of 17.5 Gy. Thirteen patients were treated for progressive/recurrent disease after surgical resection or whole brain radiotherapy (WBRT). Fifty-one patients were treated with frameless SRS as an an adjunct to initial treatment. Of the total treated, 17 were treated with SRS alone, 20 were treated with WBRT plus SRS, 16 were treated with surgical resection plus SRS, and the remaining 11 were treated with surgical resection plus WBRT plus SRS. RESULTS: With a median actuarial follow-up period of 8.2 months, ultimate local control was 88%. The median time to progression was 8.1 months. The median overall survival was 8.7 months. Of the 17 patients treated with SRS alone, 86% had ultimate local control with mean overall survival of 7.1 months. Of the 13 patients who received surgical resection plus SRS without WBRT as primary treatment, there was 85% ultimate local control with an overall survival of 10.3 months. Three patients treated with initial surgery alone had recurrence treated with SRS 2-3 months after resection. All these patients obtained local control and median survival was >10 months. Of the 13 patients who received WBRT followed by SRS as boost treatment, 92% had local control and mean overall survival was 7.3 months. Of 7 patients who received SRS after recurrence after WBRT, 100% had local control with median survival of 8.2 months. For 8 patients who received surgery followed by WBRT and SRS, local control was 50%; however, ultimate intracranial control was achieved in 7 of 8 patients with repeat SRS and surgical resection. The overall survival in this group of patients was 14.7 months. No patient had a serious (Grade 3 or higher) complication requiring intervention. CONCLUSIONS: Frameless optically guided radiosurgery is less invasive, can be performed as a standard radiotherapy-based simulation procedure, and maintains submillimetric accuracy. Our initial results with frameless SRS for metastatic disease suggest survival times and local control (88%) eqiuvalent to frame-based methodologies. Practical noninvasive delivery makes treatment and potential retreatment to avoid WBRT more feasible. PMID- 15817353 TI - Treatment of bone metastases with palliative radiotherapy: patients' treatment preferences. AB - PURPOSE: To determine the proportion of patients undergoing palliative radiotherapy (RT) for bone pain who would like to participate in the decision making process, and to determine their choice of palliative RT regimen (2000 cGy in five fractions vs. 800 cGy in one fraction) for painful bone metastases. METHODS AND MATERIALS: Eligible patients were approached and all patients agreeing to participate provided written informed consent. Patients' decisional preferences were studied using a five-statement preference instrument. A decision board was used to help patients decide their preferred palliative RT regimen. Factors influencing patients' choices were studied using a visual analog scale. RESULTS: A total of 101 patients were enrolled in the study (55 women and 46 men). The preferences for decision-making were as follows: 30 active, 47 collaborative, and 24 passive. Most (55 [76%] of 72) patients favored one fraction of palliative RT (95% confidence interval, 65-86%). Patients were more likely to select the 800 cGy in one fraction because of the convenience of the treatment plan (odds ratio, 1.024; 95% confidence interval, 1.004-1044) but were less likely to choose it because of the chance of bone fracture (odds ratio, 0.973; 95% confidence interval, 0.947-1.000) compared with 2000 cGy in five fractions. CONCLUSION: Most participating patients preferred to decide either by themselves or with the radiation oncologists which treatment option they preferred. An 800-cGy-in-one-fraction regimen was favored, independent of the treated site. The convenience of the treatment plan and the likelihood of bone fracture were the most important factors influencing patients' choice. PMID- 15817354 TI - The bronchiolar epithelium as a prominent source of pro-inflammatory cytokines after lung irradiation. AB - PURPOSE: To study in detail the temporal and spatial release of the pro inflammatory cytokines tumor necrosis factor alpha, interleukin (IL)-1alpha, and IL-6 in the lung tissue of C57BL/6 mice after thoracic irradiation with 12 Gy. METHODS AND MATERIALS: C57BL/6J mice were exposed to either sham irradiation or a single fraction of 12 Gy delivered to the thorax. Treated and sham-irradiated control mice were killed at 0.5 h, 1 h, 3 h, 6 h, 12 h, 24 h, 48 h, 72 h, 1 week, 2 weeks, 4 weeks, 8 weeks, 16 weeks, and 24 weeks post-irradiation (p.i.). Real time multiplex reverse transcriptase polymerase chain reaction was established to evaluate the relative messenger RNA (mRNA) expression of TNF-alpha, IL-1alpha, and IL-6 in the lung tissue of the mice (compared with nonirradiated lung tissue). Immunohistochemical detection methods (alkaline phosphatase anti alkaline phosphatase, avidin-biotin-complex [ABC]) and automated image analysis were used to quantify the protein expression of TNF-alpha, IL-1alpha, and IL-6 in the lung tissue (percentage of the positively stained area). RESULTS: Radiation induced release of the pro-inflammatory cytokines TNF-alpha, IL-1alpha, and IL-6 in the lung tissue was detectable within the first hours after thoracic irradiation. We observed statistically significant up-regulations for TNF-alpha at 1 h p.i. on mRNA (4.99 +/- 1.60) and at 6 h p.i. on protein level (7.23% +/- 1.67%), for IL-1alpha at 6 h p.i. on mRNA (11.03 +/- 0.77) and at 12 h p.i. on protein level (27.58% +/- 11.06%), for IL-6 at 6 h p.i. on mRNA (6.0 +/- 3.76) and at 12 h p.i. on protein level (7.12% +/- 1.93%). With immunohistochemistry, we could clearly demonstrate that the bronchiolar epithelium is the most prominent source of these inflammatory cytokines in the first hours after lung irradiation. During the stage of acute pneumonitis, the bronchiolar epithelium, as well as inflammatory cells in the lung interstitium, produced high amounts of TNF-alpha (with the maximal value at 4 weeks p.i.: 9.47% +/- 1.78%), IL-1alpha (with the peak value at 8 weeks p.i.: 14.76% +/- 7.77%), and IL-6 (with the peak value at 8 weeks p.i.: 4.28% +/- 1.33%). CONCLUSIONS: In the present study we have clearly demonstrated the immediate expression of the pro-inflammatory cytokines TNF-alpha, IL-1alpha, and IL-6 in the bronchiolar epithelium in the first hours after lung irradiation. A second, long-lasting release of these cytokines by the bronchiolar and alveolar epithelium, as well as by inflammatory cells, was observed at the onset of acute pneumonitis. Therefore, we postulate that lung irradiation causes immediate epithelial reaction, with the bronchiolar epithelium becoming a significant source of pro-inflammatory cytokines capable of promoting inflammation through recruitment and activation of inflammatory cells. PMID- 15817355 TI - Assessment of regional tumor hypoxia using 18F-fluoromisonidazole and 64Cu(II) diacetyl-bis(N4-methylthiosemicarbazone) positron emission tomography: Comparative study featuring microPET imaging, Po2 probe measurement, autoradiography, and fluorescent microscopy in the R3327-AT and FaDu rat tumor models. AB - PURPOSE: To compare two potential positron emission tomography (PET) tracers of tumor hypoxia in an animal model. METHODS AND MATERIALS: The purported hypoxia imaging agents (18)F-fluoromisonidazole (FMISO) and (64)Cu(II)-diacetyl-bis(N4 methylthiosemicarbazone) (Cu-ATSM) were compared by serial microPET imaging of Fisher-Copenhagen rats bearing the R3327-AT anaplastic rat prostate tumor. Probe measurements of intratumoral Po(2) were compared with the image data. At the microscopic level, the relationship between the spatial distributions of (64)Cu (assessed by digital autoradiography) and tumor hypoxia (assessed by immunofluorescent detection of pimonidazole) was examined. (18)F-FMISO and (64)Cu ATSM microPET images were also acquired in nude rats bearing xenografts derived from the human squamous cell carcinoma cell line, FaDu. RESULTS: In R3327-AT tumors, the intratumoral distribution of (18)F-FMISO remained relatively constant 1-4 h after injection. However, that of (64)Cu-ATSM displayed a significant temporal evolution for 0.5-20 h after injection in most tumors. In general, only when (64)Cu-ATSM was imaged at later times (16-20 h after injection) did it correspond to the distribution of (18)F-FMISO. Oxygen probe measurements were broadly consistent with (18)F-FMISO and late (64)Cu-ATSM images but not with early (64)Cu-ATSM images. At the microscopic level, a negative correlation was found between tumor hypoxia and (64)Cu distribution when assessed at early times and a positive correlation when assessed at later times. For the FaDu tumor model, the early and late (64)Cu-ATSM microPET images were similar and were in general concordance with the (18)F-FMISO scans. CONCLUSION: The difference in behavior between the R3327-AT and FaDu tumor models suggests a tumor-specific dependence of Cu-ATSM uptake and retention under hypoxic conditions. PMID- 15817356 TI - Increased oxygenation of intracranial tumors by efaproxyn (efaproxiral), an allosteric hemoglobin modifier: In vivo EPR oximetry study. AB - PURPOSE: To determine quantitatively the changes in oxygenation of intracranial tumors induced by efaproxiral, an allosteric hemoglobin modifier. Efaproxiral reduces hemoglobin-oxygen binding affinity, which facilitates oxygen release from hemoglobin into surrounding tissues and potentially increases the pO(2) of the tumors. METHODS AND MATERIALS: The study was performed on 10 male Fisher 344 rats with 9L intracranial tumors. Electron paramagnetic resonance (EPR) oximetry was used to measure quantitatively the changes in the pO(2) in the tumors. Lithium phthalocyanine (LiPc) crystals were implanted in the tumors and in the normal brain tissue in the opposite hemispheres. We monitored the cerebral pO(2) starting 7 to 10 days after the tumor cells were implanted. NMR imaging determined the position and size of tumor in the brain. After an initial baseline EPR measurement, efaproxiral (150 mg/kg) was injected intravenously over 15 minutes, and measurements of tumor and normal brain oxygen tension were made alternately at 10-minute intervals for the next 60 minutes; the procedure was repeated for 6 consecutive days. RESULTS: Efaproxiral significantly increased the pO(2) of both the intracranial tumors and the normal brain tissue on all days. The maximum increase was reached at 52.9 to 59.7 minutes and 54.1 to 63.2 minutes after injection, respectively. The pO(2) returned to baseline values at 106 to 126.5 minutes after treatment. The maximum tumor and normal tissue pO(2) values achieved after efaproxiral treatment from Day 1 through Day 6 ranged from 139.7 to 197.7 mm Hg and 103.0 to 135.9 mm Hg, respectively. The maximum increase in tumor tissue pO(2) values from Day 2 to Day 5 was greater than the maximum increase in normal tissue pO(2). CONCLUSION: We obtained quantitative data on the timing and extent of efaproxiral-induced changes in the pO(2) of intracerebral 9L tumors. These results illustrate a unique and useful capability of in vivo EPR oximetry to obtain repeated noninvasive measurements of tumor oxygenation over a number of days. The information on the dynamics of tumor pO(2) after efaproxiral administration illustrates the ability of efaproxiral to increase intracranial tumor oxygenation. PMID- 15817357 TI - Estimation of radiation-induced cancer from three-dimensional dose distributions: Concept of organ equivalent dose. AB - PURPOSE: Estimates of secondary cancer risk after radiotherapy are becoming more important for comparative treatment planning. Modern treatment planning systems provide accurate three-dimensional dose distributions for each individual patient. These data open up new possibilities for more precise estimates of secondary cancer incidence rates in the irradiated organs. We report a new method to estimate organ-specific radiation-induced cancer incidence rates. The concept of an organ equivalent dose (OED) for radiation-induced cancer assumes that any two dose distributions in an organ are equivalent if they cause the same radiation-induced cancer incidence. METHODS AND MATERIALS: The two operational parameters of the OED concept are the organ-specific cancer incidence rate at low doses, which is taken from the data of the atomic bomb survivors, and cell sterilization at higher doses. The effect of cell sterilization in various organs was estimated by analyzing the secondary cancer incidence data of patients with Hodgkin's disease who were treated with radiotherapy in between 1962 and 1993. The radiotherapy plans used at the time the patients had been treated were reconstructed on a fully segmented whole body CT scan. The dose distributions were calculated in individual organs for which cancer incidence data were available. The model parameter that described cell sterilization was obtained by analyzing the dose and cancer incidence rates for the individual organs. RESULTS: We found organ-specific cell radiosensitivities that varied from 0.017 for the mouth and pharynx up to 1.592 for the bladder. Using the two model parameters (organ-specific cancer incidence rate and the parameter characterizing cell sterilization), the OED concept can be applied to any three-dimensional dose distribution to analyze cancer incidence. CONCLUSION: We believe that the concept of OED presented in this investigation represents a first step in assessing the potential risk of secondary cancer induction after the clinical application of radiotherapy. PMID- 15817358 TI - Preferential action of arsenic trioxide in solid-tumor microenvironment enhances radiation therapy. AB - PURPOSE: To investigate the effect of arsenic trioxide, Trisenox (TNX), on primary cultures of endothelial cells and tumor tissue under varying pH and pO(2) environments and the effects of combined TNX and radiation therapy on experimental tumors. METHODS AND MATERIALS: Human dermal microvascular endothelial cells were cultured in vitro and exposed to TNX under various combinations of aerobic, hypoxic, neutral, or acidic conditions, and levels of activated JNK MAP kinase were assessed by Western blotting. FSaII fibrosarcoma cells grown in the hind limb of female C3H mice were used to study the effect of TNX on tumor blood perfusion and oxygenation. The tumor-growth delay after a single or fractionated irradiation with or without TNX treatment was assessed. RESULTS: A single intraperitoneal injection of 8 mg/kg TNX reduced the blood perfusion in FSaII tumors by 53% at 2 hours after injection. To increase the oxygenation of the tumor vasculature during TNX treatment, some animals were allowed to breathe carbogen (95% O(2)/5% CO(2)). Carbogen breathing alone for 2 hours reduced tumor perfusion by 33%. When carbogen breathing was begun immediately after TNX injection, no further reduction occurred in tumor blood perfusion at 2 hours after injection. In vitro, TNX exposure increased activity JNK MAP kinase preferentially in endothelial cells cultured in an acidic or hypoxic environment. In vivo, the median oxygenation in FSaII tumors measured at 3 or 5 days after TNX injection was found to be significantly elevated compared with control tumors. Subsequently, radiation-induced tumor-growth delay was synergistically increased when radiation and TNX injection were fractionated at 3 day or 5-day intervals. CONCLUSIONS: Trisenox has novel vascular-damaging properties, preferentially against endothelium in regions of low pH or pO(2), which leads to tumor cell death and enhancement of the response of tumors to radiotherapy. PMID- 15817359 TI - Alteration of cancer pain-related signals by radiation: proteomic analysis in an animal model with cancer bone invasion. AB - PURPOSE: Although radiotherapy is highly effective in relieving bone pain due to cancer invasion, its mechanism remains unclear. The aim of this study was to explore this mechanism in an animal model system. METHODS AND MATERIALS: A hind paw model of cancer pain was developed by transplanting a murine hepatocarcinoma, HCa-1, into the periosteal membrane of the foot dorsum of C3H/HeJ mice. Bone invasion from HCa-1 was histopathologically confirmed from sequential tumor sampling. For three experimental groups, a control (N), tumor without radiation (T), and tumor with radiation (TR), the development and level of pain were objectively examined in mice with a growing tumor by assessing pain-associated behavior. The differential expression of pain-related signals in the spinal cord was analyzed by proteomic analysis using high-resolution two-dimensional gel electrophoresis (2-DE) and mass spectrometry, and those of proteins by Western blotting. The pain-mediating neurotransmitters in the spinal cord were also examined by immunohistochemical staining for calcitonin gene-related peptide (CGRP) and substance P. RESULTS: In the histopathologic examinations, bone invasion from HCa-1 was seen from Day 7 and was evident at Day 14 after transplantation, and measurable pain-associated behaviors were developed from Day 7. After 25 Gy of radiation to the tumors, the objective level of pain in the TR group decreased, with higher thresholds to mechanical and thermal stimulation than in the T group. From the 2-DE of spinal cord, 107 spots were identified; 12 proteins were changed more than fivefold because of tumor formation but then reversed after radiation in the tumor-bearing mice. The proteins involved included secretagogin, syntenin, P2X purinoreceptor 6 (P2X6), and Ca(2+)/Calmodulin-dependent protein kinase 1 (CaM kinase 1), the functions of which have been known to be involved in the Ca(2+)-signaling cascade, ATP mediated fast synaptic transmission, or control of vesicular trafficking. Validations using Western blotting were successful for the CaM kinase and P2X6. In immunohistochemical staining of the spinal cord, a significant decrease after irradiation was shown in the expression of CGRP, but not in substance P. CONCLUSIONS: We developed a novel model for bone pain due to cancer invasion, which was confirmed by histopathologic examination and measurable pain-associated behaviors. Radiotherapy decreased the objective level of pain. The underlying mechanism seems to be related to the Ca(2+)-signaling cascade or control of vesicular trafficking. PMID- 15817360 TI - Four-dimensional image-based treatment planning: Target volume segmentation and dose calculation in the presence of respiratory motion. AB - PURPOSE: To describe approaches to four-dimensional (4D) treatment planning, including acquisition of 4D-CT scans, target delineation of spatio-temporal image data sets, 4D dose calculations, and their analysis. METHODS AND MATERIALS: The study included patients with thoracic and hepatocellular tumors. Specialized tools were developed to facilitate visualization, segmentation, and analysis of 4D-CT data: maximum intensity volume to define the extent of lung tumor motion, a 4D browser to examine and dynamically assess the 4D data sets, dose calculations, including respiratory motion, and deformable registration to combine the dose distributions at different points. RESULTS: Four-dimensional CT was used to visualize and quantitatively assess respiratory target motion. The gross target volume contours derived from light breathing scans showed significant differences compared with those extracted from 4D-CT. Evaluation of deformable registration using difference images of original and deformed anatomic maps suggested the algorithm is functionally useful. Thus, calculation of effective dose distributions, including respiratory motion, was implemented. CONCLUSION: Tools and methods to use 4D-CT data for treatment planning in the presence of respiratory motion have been developed and applied to several case studies. The process of 4D-CT-based treatment planning has been implemented, and technical barriers for its routine use have been identified. PMID- 15817361 TI - The correlation between internal and external markers for abdominal tumors: implications for respiratory gating. AB - PURPOSE: The correlation of the respiratory motion of external patient markers and abdominal tumors was examined. Data of this type are important for image guided therapy techniques, such as respiratory gating, that monitor the movement of external fiducials. METHODS AND MATERIALS: Fluoroscopy sessions for 4 patients with internal, radiopaque tumor fiducial clips were analyzed by computer vision techniques. The motion of the internal clips and the external markers placed on the patient's abdominal skin surface were quantified and correlated. RESULTS: In general, the motion of the tumor and external markers were well correlated. The maximum amount of peak-to-peak craniocaudal tumor motion was 2.5 cm. The ratio of tumor motion to external-marker motion ranged from 0.85 to 7.1. The variation in tumor position for a given external-marker position ranged from 2 to 9 mm. The period of the breathing cycle ranged from 2.7 to 4.5 seconds, and the frequency patterns for both the tumor and the external markers were similar. CONCLUSIONS: Although tumor motion generally correlated well with external fiducial marker motion, relatively large underlying tumor motion can occur compared with external marker motion and variations in the tumor position for a given marker position. Treatment margins should be determined on the basis of a detailed understanding of tumor motion, as opposed to relying only on external-marker information. PMID- 15817362 TI - Real-time monitoring of a digestive tract marker to reduce adverse effects of moving organs at risk (OAR) in radiotherapy for thoracic and abdominal tumors. AB - PURPOSE: To evaluate the feasibility of real-time monitoring of a fiducial marker in/near the digestive tract and to analyze the motion of organs at risk to determine a reasonable internal margin. METHODS AND MATERIALS: We developed two methods to insert a fiducial marker into/near the digestive tract adjacent to the target volume. One method involves an intraoperative insertion technique, and the other involves endoscopic insertion into the submucosal layer of the normal digestive tract. A fluoroscopic real-time tumor-tracking radiotherapy system was used to monitor the marker. RESULTS: Fourteen markers (2 in the mediastinum and 12 in the abdomen) were implanted intraoperatively in 14 patients with no apparent migration. Seventeen of 20 markers (13/14 in the esophagus, 1/2 in the stomach, and 3/4 in the duodenum) in 18 patients were implanted using endoscopy without dropping. No symptomatic adverse effects related to insertion were observed. The mean/standard deviation of the range of motion of the esophagus was 3.5/1.8, 8.3/3.8, and 4.0/2.6 mm for lateral, craniocaudal and anteroposterior directions, respectively, in patients with intrafractional tumor motion less than 1.0 cm. CONCLUSION: Both intraoperative and endoscopic insertions of a fiducial marker into/near the digestive tract for monitoring of organs at risk were feasible. The margin for internal motion can be individualized using this system. PMID- 15817363 TI - Treatment planning of stereotactic radiotherapy for solitary lung tumor. AB - PURPOSE: To analyze the stereotactic radiotherapy (SRT) plans in terms of internal target volume (ITV) and organs at risk (OARs). METHODS AND MATERIALS: Treatment planning and dose distributions were analyzed using dose-volume histograms (DVHs) of ITV and OARs in 37 patients, who were treated for a solitary lung tumor with SRT. The stereotactic body frame (SBF) was used for immobilization and accurate setup. Prescription dose was 48 Gy in four fractions at the isocenter. RESULTS: Use of SBF limits the extent of the noncoplanar beam directions to prevent a collision with the Linac gantry. DVH analyses showed that the homogeneity index, defined as the ratio of maximum and minimum dose to ITV, ranged from 1.03 to 1.25 (mean, 1.12). The volume irradiated with 20 Gy or more (V(20)) of the lung ranged from 0.3 to 11.6% (mean, 4.4%) of the whole lung volume. The maximum dose to the other OARs ranged from 0 to 11.8 Gy (mean, 0.5 2.7) per fraction. No clinically significant complications were encountered. CONCLUSIONS: Despite the limitation of the beam arrangement, a homogeneous target dose distribution, while avoiding high doses to normal tissues, was obtained. PMID- 15817364 TI - Impact of IMRT and leaf width on stereotactic body radiotherapy of liver and lung lesions. AB - PURPOSE: The present study explored the impact of intensity-modulated radiotherapy (IMRT) on stereotactic body RT (SBRT) of liver and lung lesions. Additionally, because target dose conformity can be affected by the leaf width of a multileaf collimator (MLC), especially for small targets and stereotactic applications, the use of a micro-MLC on "uniform intensity" conformal and intensity-modulated SBRT was evaluated. METHODS AND MATERIALS: The present study included 10 patients treated previously with SBRT in our institution (seven lung and three liver lesions). All patients were treated with 3 x 12 Gy prescribed to the 65% isodose level. The actual MLC-based conformal treatment plan served as the standard for additional comparison. In total, seven alternative treatment plans were made for each patient: a standard (actual) plan and an IMRT plan, both calculated with Helax TMS (Nucletron) using a pencil beam model; and a recalculated standard and a recalculated IMRT plan on Helax TMS using a point dose kernel approach. These four treatment plans were based on a standard MLC with 1-cm leaf width. Additionally, the following micro-MLC (central leaf width 3 mm)-based treatment plans were calculated with the BrainSCAN (BrainLAB) system: standard, IMRT, and dynamic arc treatments. For each treatment plan, various target parameters (conformity, coverage, mean, maximal, and minimal target dose, equivalent uniform doses, and dose-volume histogram), as well as organs at risk parameters (3 Gy and 6 Gy volume, mean dose, dose-volume histogram) were evaluated. Finally, treatment efficiency was estimated from monitor units and the number of segments for IMRT solutions. RESULTS: For both treatment planning systems, no significant difference could be observed in terms of target conformity between the standard and IMRT dose distributions. All dose distributions obtained with the micro-MLC showed significantly better conformity values compared with the standard and IMRT plans using a regular MLC. Dynamic arc plans were characterized by the steepest dose gradient and thus the smallest V(6 Gy) values, which were on average 7% smaller than the standard plans and 20% lower than the IMRT plans. Although the Helax TMS IMRT plans show about 18% more monitor units than the standard plan, BrainSCAN IMRT plans require approximately twice the number of monitor units relative to the standard plan. All treatment plans optimized with a pencil beam model but recalculated with a superposition method showed significant qualitative, as well as quantitative, differences, especially with respect to conformity and the dose to organs at risk. CONCLUSION: Standard conformal treatment techniques for SBRT could not be improved with inversely planned IMRT approaches. Dose calculation algorithms applied in optimization modules for IMRT applications in the thoracic region need to be based on the most accurate dose calculation algorithms, especially when using higher energy photon beams. PMID- 15817365 TI - Quantifying IOHDR brachytherapy underdosage resulting from an incomplete scatter environment. AB - PURPOSE: Most brachytherapy planning systems are based on a dose calculation algorithm that assumes an infinite scatter environment surrounding the target volume and applicator. Dosimetric errors from this assumption are negligible. However, in intraoperative high-dose-rate brachytherapy (IOHDR) where treatment catheters are typically laid either directly on a tumor bed or within applicators that may have little or no scatter material above them, the lack of scatter from one side of the applicator can result in underdosage during treatment. This study was carried out to investigate the magnitude of this underdosage. METHODS: IOHDR treatment geometries were simulated using a solid water phantom beneath an applicator with varying amounts of bolus material on the top and sides of the applicator to account for missing tissue. Treatment plans were developed for 3 different treatment surface areas (4 x 4, 7 x 7, 12 x 12 cm(2)), each with prescription points located at 3 distances (0.5 cm, 1.0 cm, and 1.5 cm) from the source dwell positions. Ionization measurements were made with a liquid-filled ionization chamber linear array with a dedicated electrometer and data acquisition system. RESULTS: Measurements showed that the magnitude of the underdosage varies from about 8% to 13% of the prescription dose as the prescription depth is increased from 0.5 cm to 1.5 cm. This treatment error was found to be independent of the irradiated area and strongly dependent on the prescription distance. Furthermore, for a given prescription depth, measurements in planes parallel to an applicator at distances up to 4.0 cm from the applicator plane showed that the dose delivery error is equal in magnitude throughout the target volume. CONCLUSION: This study demonstrates the magnitude of underdosage in IOHDR treatments delivered in a geometry that may not result in a full scatter environment around the applicator. This implies that the target volume and, specifically, the prescription depth (tumor bed) may get a dose significantly less than prescribed. It might be clinically relevant to correct for this inaccuracy. PMID- 15817366 TI - An ultrasonic device for source to skin surface distance measurement in patient setup. AB - PURPOSE: To develop an ultrasound-based source to skin surface distance (SSD) measurement technique and device for patient setup and test its feasibility and accuracy. METHODS AND MATERIALS: The ultrasonic SSD measurement device (USD) prototype consists of two main parts: a probe plate with an ultrasonic transducer in the center and a control unit that displays the SSD in millimeters. The probe plate can be slid into the block tray accessory slot of any treatment machine at the time of the SSD measurement. The probe plate contains an ultrasonic transducer as both the source and the detector for measuring the distance between the transducer and the target surfaces on the basis of an echo-detecting technique. The device was calibrated by a mechanical ruler with an accuracy of 0.01 mm and corrected by an offset of 601.7 mm, which is the distance from the radiation source to the ultrasonic transducer surface for the Siemens Primus linear accelerator (Linac). The ultrasound device provided digital readout with an accuracy of +/-0.1 mm for a flat surface after calibration. The SSD measurement experiments were done with the USD, an optical distance indicator (ODI), and an AKTINA 53-104 Mechanical Front Pointer (FP) on a Siemens Primus Linac with a full-sized female phantom. Ten measurements were carried out at each gantry angle of 0 degrees , 52 degrees , 85 degrees , 90 degrees , and 227 degrees for anatomic locations of head, thorax, breast, and pelvis, to obtain the average values and standard deviations. RESULTS: The comparison study with the ODI and FP showed that the USD had an accuracy of less than +/-1.0 mm and that USD measurements had the minimum standard deviations among the three methods; therefore, USD gave more consistent and accurate readouts for SSD measurement. When considering the FP as a reference, the USD yields smaller deviations than the ODI for all measured locations (less than +/-2 mm). The variation of USD digital readout with a room temperature change of +/-2 degrees C is +/-0.1 mm, which is sufficiently accurate for SSD measurement. CONCLUSIONS: The USD method has the following advantages. First, it decreases patient setup time by avoiding problems related to the blocking of the device by the patient or by the immobilization device. Second, it is more accurate than the other two methods currently used, as the test data show. Last, the digital readout eliminates the possibility of human reading error associated with the visual scales. PMID- 15817367 TI - Error in the delivery of radiation therapy: results of a quality assurance review. AB - PURPOSE: To examine error rates in the delivery of radiation therapy (RT), technical factors associated with RT errors, and the influence of a quality improvement intervention on the RT error rate. METHODS AND MATERIALS: We undertook a review of all RT errors that occurred at the Princess Margaret Hospital (Toronto) from January 1, 1997, to December 31, 2002. Errors were identified according to incident report forms that were completed at the time the error occurred. Error rates were calculated per patient, per treated volume (>/=1 volume per patient), and per fraction delivered. The association between tumor site and error was analyzed. Logistic regression was used to examine the association between technical factors and the risk of error. RESULTS: Over the study interval, there were 555 errors among 28,136 patient treatments delivered (error rate per patient = 1.97%, 95% confidence interval [CI], 1.81-2.14%) and among 43,302 treated volumes (error rate per volume = 1.28%, 95% CI, 1.18-1.39%). The proportion of fractions with errors from July 1, 2000, to December 31, 2002, was 0.29% (95% CI, 0.27-0.32%). Patients with sarcoma or head-and-neck tumors experienced error rates significantly higher than average (5.54% and 4.58%, respectively); however, when the number of treated volumes was taken into account, the head-and-neck error rate was no longer higher than average (1.43%). The use of accessories was associated with an increased risk of error, and internal wedges were more likely to be associated with an error than external wedges (relative risk = 2.04; 95% CI, 1.11-3.77%). Eighty-seven errors (15.6%) were directly attributed to incorrect programming of the "record and verify" system. Changes to planning and treatment processes aimed at reducing errors within the head-and-neck site group produced a substantial reduction in the error rate. CONCLUSIONS: Errors in the delivery of RT are uncommon and usually of little clinical significance. Patient subgroups and technical factors associated with errors can be identified. The introduction of new technology can produce new ways for errors to occur, necessitating ongoing evaluation of RT errors for quality assurance. Modifications to processes of care can produce important reductions in error rates. PMID- 15817368 TI - The effect of finite patient dimensions and tissue inhomogeneities on dosimetry planning of 192Ir HDR breast brachytherapy: a Monte Carlo dose verification study. AB - PURPOSE: To evaluate the accuracy of clinical dosimetry planning using commercially available treatment planning systems in (192)Ir high-dose-rate (HDR) breast brachytherapy, with emphasis on skin dose, in view of potential uncertainties owing to the patient finite dimensions and the presence of the lung. METHODS AND MATERIALS: A patient-equivalent mathematical phantom was constructed on the basis of the patient computed tomography scan used in the clinical treatment planning procedure. The actual treatment plan delivered to the patient, involving an implant of six plastic catheters and 26 programmed source dwell positions, was simulated by means of the Monte Carlo method. Results are compared with corresponding dose calculations of a commercially available treatment planning system in the form of prescribed dose percentage isodose contours and cumulative dose-volume histograms. RESULTS: The comparison of Monte Carlo results and treatment planning system calculations revealed that all percentage isodose contours greater than 60% of the prescribed dose are not affected by the finite breast dimensions or the presence of the lung. Treatment planning system calculations overestimate dose in the lung as well as lower isodose contours at points lying both close to the breast or lung surface and relatively away from the implant. In particular, skin dose is overestimated by 5% in the central breast region and within 10% at all other points. CONCLUSIONS: Dose-volume histogram and all other relevant planning quality indices for the planning target volume calculated by the treatment planning system are credible. Skin and lung dose calculations by the treatment planning system can be thought of as a conservative approach in view of the reported dose overestimation. PMID- 15817369 TI - In vivo measurements with MOSFET detectors in oropharynx and nasopharynx intensity-modulated radiation therapy. AB - PURPOSE: To evaluate the feasibility of in vivo measurements with metal oxide semiconductor field effect transistor (MOSFET) dosimeters for oropharynx and nasopharynx intensity-modulated radiation therapy (IMRT). METHODS AND MATERIALS: During a 1-year period, in vivo measurements of the dose delivered to one or two points of the oral cavity by IMRT were obtained with MOSFET dosimeters. Measurements were obtained during each session of 48 treatment plans for 21 patients, all of whom were fitted with a custom-made mouth plate. Calculated and measured values were compared. RESULTS: A total of 344 and 452 measurements were performed for the right and left sides, respectively, of the oral cavity. Seventy percent of the discrepancies between calculated and measured values were within +/-5%. Uncertainties were due to interfraction patient positions, intrafraction patient movements, and interfraction MOSFET positions. Nevertheless, the discrepancies between the measured and calculated means were within +/-5% for 92% and 95% of the right and left sides, respectively. Comparison of these discrepancies and the discrepancies between calculated values and measurements made on a phantom revealed that all differences were within +/-5%. CONCLUSION: Our experience demonstrates the feasibility of in vivo measurements with MOSFET dosimeters for oropharynx and nasopharynx IMRT. PMID- 15817370 TI - Tumor hypoxia versus blood flow: In regard to Lehtio et al. (Int J Radiat Oncol Biol Phys 2004;59:971-982). PMID- 15817372 TI - Mitochondrial fusion and fission in the control of apoptosis. AB - Mitochondrial outer membrane permeabilization (MOMP) determines the point-of-no return of most if not all signal-transduction cascades leading to cell death. It has been postulated that the molecular mechanism leading to MOMP could depend on the activation of the mitochondrial fission machinery mediated by proteins from the dynamin superfamily. However, recent work suggests that, depending on the specific apoptosis induction pathway, mitochondrial fission can occur independently or downstream from MOMP. Moreover, fragmentation of the mitochondrial network can inhibit MOMP and apoptosis in response to a particular range of lethal stimuli, namely those relying on Ca(2+) waves. Failure to transmit the Ca(2+) wave through disconnected mitochondria then interrupts the propagation of the pro-apoptotic signal. Thus, mitochondrial fission can either enhance or reduce the probability of MOMP and consequent cell death, depending on the initial lethal stimulus. PMID- 15817373 TI - Neuronal microtubules: when the MAP is the roadblock. AB - Recent studies shed new light on a potential cascade of events by which neurological diseases such as Alzheimer's lead to axonal degeneration. In this model, the pathology starts with an elevation in microtubule-associated proteins (MAPs) such as tau. This renders the microtubules less accessible to motor proteins, which impairs their capacity to sustain anterograde axonal transport of proteins and organelles. In response, the neuron hyperphosphorylates tau so that it dissociates from the microtubules. Unfortunately, the hyperphosphorylated tau forms abnormal filaments that are deleterious to the axon, and the tau-depleted microtubules become highly sensitive to microtubule-severing proteins such as katanin. PMID- 15817374 TI - Macrophage fusion: are somatic and cancer cells possible partners? AB - Macrophages are present in all tissues and can fuse with themselves to differentiate into multinucleate osteoclasts or giant cells that play a central role in osteoporosis and chronic inflammatory diseases, respectively. Yet, the mechanism by which they fuse remains uncharacterized. The macrophage fusion receptor (MFR) and its ligand CD47 might mediate homotypic fusion of macrophages and allow for their recognition as 'self' before fusion. Although a novel process and controversial idea, macrophages might exploit a similar mechanism for fusion with somatic cells or tumor cells, with resultant organ repair or metastasis, respectively. Hence, macrophages might be the 'double-edged swords' of tissues. PMID- 15817375 TI - Birth of a nucleolus: the evolution of nucleolar compartments. AB - In eukaryotes, ribosome synthesis largely takes place in a specialized nuclear domain - the nucleolus. It has recently become apparent that this organelle is involved in the biogenesis of most cellular ribonucleoprotein particles (RNPs), as well as in cell-cycle regulation, making it central to gene expression. The field has traditionally acknowledged that each nucleolus is organized in three morphologically distinct compartments. Here, however, we discuss our view that in fact many eukaryotes have bipartite nucleoli. We propose that, during evolution, a third nucleolar compartment emerged at the transition between the anamniotes and the amniotes, following a substantial increase in size of the rDNA intergenic region. We believe that these conclusions have important implications for understanding the structure-function relationships within this key cellular organelle. PMID- 15817376 TI - The stress and strain of cytokinesis. AB - The ultimate goal of all signaling pathways in cytokinesis is to control the mechanical separation of the mother cell into two daughter cells. Because of the intrinsic mechanical nature of cytokinesis, it is essential to understand fully how cell shapes and the material properties of the cell are generated, how these shapes and material properties create force, and how motor proteins such as myosin-II modify the system to achieve successful cytokinesis. In this review (which is part of the Cytokinesis series), we discuss the relevant physical properties of cells, how these properties are measured and the basic models that are used to understand cell mechanics. Finally, we present our current understanding of how cytokinesis mechanics work. PMID- 15817377 TI - Fluorescence microscopy with super-resolved optical sections. AB - The fluorescence microscope, especially its confocal variant, has become a standard tool in cell biology research for delivering 3D-images of intact cells. However, the resolution of any standard optical microscope is at least 3 times poorer along the axis of the lens that in its focal plane. Here, we review principles and applications of an emerging family of fluorescence microscopes, such as 4Pi microscopes, which improve axial resolution by a factor of seven by employing two opposing lenses. Noninvasive axial sections of 80-160 nm thickness deliver more faithful 3D-images of subcellular features, providing a new opportunity to significantly enhance our understanding of cellular structure and function. PMID- 15817378 TI - Synaptic contact dynamics controlled by cadherin and catenins. AB - A synapse is the connection between neurons that joins an axon of one neuron to the dendrite of another. One class of synapses is formed at the contact point between an axon and a small protrusion from a dendrite, called a dendritic spine. These spines are motile and deformable, which indicates that synaptic functions are controlled, at least in part, by their morphological changes. Recent studies show that the cadherin cell-adhesion molecules and their cytoplasmic partners, catenins, can modulate axon-spine contacts in a manner that responds to neural activity. These observations indicate that cadherins, which are essential for general cell-cell adhesion, also play a role in the control of synaptic dynamics. PMID- 15817379 TI - The building blocks for basolateral vesicles in polarized epithelial cells. AB - After the discovery of basolateral sorting signals for polarized delivery in epithelial cells in the early 1990s, it was only about a decade later that the epithelial-cell-specific sorting adaptor AP-1B was discovered. AP-1B decodes a subclass of basolateral sorting signals and localizes to the recycling endosomes as opposed to the trans-Golgi network, suggesting that this is its major site of action. Furthermore, AP-1B does not simply select its cargo but also facilitates the recruitment of the exocyst complex needed for subsequent fusion with the plasma membrane. This review discusses our current knowledge of AP-1B function in cargo sorting to the basolateral membrane and its impact on our understanding of the similarities and differences between AP-1B-minus fibroblasts and AP-1B positive epithelial cells. PMID- 15817380 TI - Host factors regulating post-integration latency of HIV. AB - A recent study has provided important clues towards the identity of the host genes that conspire to promote post-integration latency of human immunodeficiency virus (HIV). Various genes controlling transcription, histone deacetylation and proteasome-mediated protein degradation have emerged as potential players. If the desired, but difficult, goal of complete virus eradication in HIV-infected patients is ever to be realized, the latent reservoir of HIV proviruses must be cleared. Understanding the molecular basis for viral latency is the key first step. PMID- 15817381 TI - Molecular gymnastics: distortion of an RNA polymerase sigma factor. AB - A recent structure obtained by nuclear magnetic resonance (NMR) spectroscopy shows that the binding of a small phage factor to the sigma(70) subunit of Escherichia coli RNA polymerase induces an unprecedented remodeling of a region of sigma(70), converting a DNA-binding helix-turn-helix into a continuous pseudohelix. This conformational change suggests how the phage factor can function both as an inhibitor and co-activator of transcription. PMID- 15817382 TI - Bacterial flagellar diversity in the post-genomic era. AB - Flagellar biosynthesis has been studied most thoroughly in laboratory strains of Escherichia coli and Salmonella enterica. However, genome sequencing has uncovered flagellar loci in distantly related bacteria. We have used homology searches to determine how far the E. coli/S. enterica paradigm can be generalised to other flagellar systems. Numerous previously unrecognized homologues of flagellar components were discovered, including novel FlgM, FlgN, FliK and FliO homologues. Homology was found between the FliK proteins and a molecular ruler, YscP, from a virulence-associated type-III secretion system. Also described is a new family of flagellar proteins, the FlhX proteins, which resemble the cytoplasmic domain of FlhB. PMID- 15817383 TI - PIRSpred: a web server for reliable HIV-1 protein-inhibitor resistance/susceptibility prediction. PMID- 15817384 TI - Bacterial observations: a rudimentary form of intelligence? AB - Genome sequencing has revealed that signal transduction in bacteria makes use of a limited number of different devices, such as two-component systems, LuxI-LuxR quorum-sensing systems, phosphodiesterases, Ser-Thr (serine-threonine) kinases, OmpR-type regulators, and sigma factor-anti-sigma factor pathways. These systems use modular proteins with a large variety of input and output domains, yet strikingly conserved transmission domains. This conservation might lead to redundancy of output function, for example, via crosstalk (i.e. phosphoryl transfer from a non-cognate sensory kinase). The number of similar devices in a single cell, particularly of the two-component type, might amount to several dozen, and most of these operate in parallel. This could bestow bacteria with cellular intelligence if the network of two-component systems in a single cell fulfils the requirements of a neural network. Testing these ideas poses a great challenge for prokaryotic systems biology. PMID- 15817385 TI - Hepatitis C virus, ER stress, and oxidative stress. AB - Hepatitis C virus (HCV) replication is associated with the endoplasmic reticulum (ER), where the virus causes stress. Cells cope with ER stress by activating an adaptive program called the unfolded protein response (UPR), which alleviates this stress by stimulating protein folding and degradation in the ER and down regulating overall protein synthesis. Recent work suggests that HCV also alters ER calcium homeostasis, inducing oxidative stress. Future progress in understanding the control that HCV exerts over the ER will provide insight into viral strategies for pathogenesis and persistence in chronically infected patients. PMID- 15817386 TI - Insights into viral transmission at the uterine-placental interface. AB - During human gestation, viruses can cause intrauterine infections associated with pregnancy complications and fetal abnormalities. The ability of viruses to spread from the infected mother to the fetus arises from the architecture of the placenta, which anchors the fetus to the uterus. Placental cytotrophoblasts differentiate, assume an endothelial phenotype, breach uterine blood vessels and form a hybrid vasculature that amplifies the maternal blood supply for fetal development. Human cytomegalovirus - the major cause of congenital disease - infects the uterine wall and the adjacent placenta, suggesting adaptation for pathogen survival in this microenvironment. Infection of villus explants and differentiating and/or invading cytotrophoblasts offers an in vitro model for studying viruses associated with prenatal infections. PMID- 15817387 TI - Export of complex cofactor-containing proteins by the bacterial Tat pathway. AB - The twin-arginine (Tat) protein translocase is a highly unusual protein transport machine that is dedicated to the movement of folded proteins across the bacterial cytoplasmic membrane. Proteins are targeted to the Tat pathway by means of N terminal signal peptides harbouring a distinctive twin-arginine motif. In the model organism Escherichia coli, many of the Tat substrates bind redox cofactors that are inserted into apo-proteins before they engage with the Tat machinery. Here we review recent advances in understanding the events involved in the coordination of cofactor insertion with the export process. Current models for Tat protein transport are also discussed. PMID- 15817388 TI - Cortactin: an Achilles' heel of the actin cytoskeleton targeted by pathogens. AB - Cortactin is an actin-binding protein and a central regulator of the actin cytoskeleton. Importantly, cortactin is also a common target exploited by microbes during infection. Its involvement in disease development is exemplified by a variety of pathogenic processes, such as pedestal formation [enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC and EHEC)], invasion (Shigella, Neisseria, Rickettsia, Chlamydia, Staphylococcus and Cryptosporidium), actin-based motility (Listeria, Shigella and vaccinia virus) and cell scattering (Helicobacter). Recent progress turns our attention to how cortactin function can be regulated by serine and tyrosine phosphorylation. This has an important impact on how pathogens abuse cortactin to modulate the architecture of the host actin cytoskeleton. PMID- 15817389 TI - Larger than Dbl: new structural insights into RhoA activation. AB - Dbl homology (DH) domains are almost always followed immediately by pleckstrin homology (PH) domains in Dbl family proteins, and these DH-PH fragments directly activate GDP-bound Rho GTPases by catalyzing the exchange of GDP for GTP. New crystal structures of the DH-PH domains from leukemia-associated Rho guanine nucleotide exchange factor (RhoGEF) and PDZ-RhoGEF bound to RhoA reveal how DH-PH domains cooperate to specifically activate Rho GTPases. PMID- 15817390 TI - The voltage-sensor structure in a voltage-gated channel. AB - A recent electron paramagnetic resonance study of KvAP, a prokaryotic voltage gated channel, in its lipid native environment has revealed the location of the transmembrane segments, the connecting loops and the relative position of the voltage-sensing charges. The results confirm that the previously reported crystal structure does not represent a native conformation and give us structural constraints that will help in determining the molecular structure of the voltage sensor. PMID- 15817391 TI - C1, see them all. AB - Recent structural analysis of crystalline beta2-chimaerin shows the central protein kinase C-like, diacylglycerol (DAG)-binding C1 domain to be masked by its intramolecular interactions with the N-terminal SH2 and GAP domains, and linker regions. A mechanism of activation has been derived from modelling of a GAP-Rac GTPase complex--the auto-inhibitory constraints are released via membrane engagement, unmasking the C1 domain to enable DAG binding and subsequent GAP stimulation of Rac GTPase catalytic activity. PMID- 15817392 TI - Centromeric chromatin makes its mark. AB - Post-translational modifications of core histones, particularly in the N terminus, seem to define different chromatin states in the genome. For instance, actively transcribed regions of euchromatin have a completely different set of 'marks' compared with silent heterochromatin. Recent analyses surprisingly demonstrate that, in addition to containing the histone H3 variant CENP-A, the centromeric chromatin that underlies the kinetochore bears a distinct combination of histone H3 modifications. PMID- 15817393 TI - Fixing the Q cycle. AB - Mitchell's key insight that all bioenergetic membranes run on the conversion of redox energy into transmembrane electrical and proton gradients took the form 30 years ago of a working model of the Q cycle of cytochrome bc1, which operates reversibly on coupled electron and proton transfers of quinone at two binding sites on opposite membrane faces. His remarkable model still stands today, but he had no structural information to provide understanding into how dangerous short circuit redox reactions were avoided. Now, it is clear that the Q cycle must be fixed with a special mechanism that avoids semiquinone-mediated short circuits. Either the redox states of the quinone electron-transfer partners double-gate the semiquinone-intermediate stability, or semiquinone is avoided altogether in concerted double-electron transfer. PMID- 15817394 TI - Multiubiquitylation by E4 enzymes: 'one size' doesn't fit all. AB - Selective protein degradation by the 26S proteasome requires the covalent attachment of several ubiquitin molecules in the form of a multiubiquitin chain. Ubiquitylation usually involves three classes of enzymes: a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2) and a ubiquitin ligase (E3). However, in some cases, multiubiquitylation requires the additional activity of certain ubiquitin-chain elongation factors. Yeast UFD2 (ubiquitin fusion degradation), for example, binds to oligoubiquitylated substrates (proteins modified by only a few ubiquitin molecules) and catalyses multiubiquitin-chain assembly in collaboration with E1, E2 and E3. Enzymes possessing this specific activity have been proposed to be termed 'E4 enzymes'. Recent studies have provided accumulating evidence that has led some researchers in the field to conclude that E4, indeed, represents a distinct and novel class of enzymes. PMID- 15817395 TI - How receptor mosaics decode transmitter signals. Possible relevance of cooperativity. AB - It has been demonstrated that receptor-receptor interactions between G-protein coupled receptors (GPCRs) occur at the plasma-membrane level. It has also been shown that clustering of GPCRs in aggregates or receptor mosaics (RMs) results in the reciprocal modulation of their binding and decoding characteristics. It is hypothesized that cooperativity plays an important part in the decoding of signals processed by RMs of GPCRs. Thus, the binding of the ligand at one receptor alters the likelihood of the same ligand binding at the next site, in the case of RMs, formed by identical receptors and/or by iso-receptors (receptors that bind the same ligand). PMID- 15817396 TI - Signalling by PI3K isoforms: insights from gene-targeted mice. AB - Phosphoinositide 3-kinases (PI3Ks) generate lipids that control a wide variety of intracellular signalling pathways. Part of this diversity in PI3K actions stems from the broad range of protein effectors of the PI3K lipids. A further layer of complexity is added by the existence of multiple isoforms of PI3K. Gene-targeting studies in the mouse have recently uncovered key roles for specific PI3K isoforms in immunity, metabolism and cardiac function. Remarkably, some of these actions do not require PI3K catalytic activity. In addition, loss-of-expression of certain PI3K genes leads to increased PI3K signalling following insulin stimulation. PI3K gene targeting has, in many cases, led to altered expression of the non-targeted PI3K subunits, making it difficult to exclude that some of the reported phenotypes result from 'knock-on' effects of PI3K gene deletion. Targeting strategies that take into account the complex interplay between members of the PI3K family will be crucial to gain a full understanding of the physiological roles of the isoforms of PI3K. PMID- 15817397 TI - Chopped, trapped or tacked--protein translocation into the IMS of mitochondria. AB - All proteins of the intermembrane space (IMS) of mitochondria are synthesized in the cytosol. The mechanisms by which these polypeptides are transported into the IMS are strikingly different from other protein-translocation processes in the cell. Recent studies suggest that IMS proteins reach their destination by three alternative principles that differ in the energy sources employed to drive the translocation reactions. The first class of proteins uses both hydrolysis of matrix ATP and the electrochemical potential of the inner membrane. The second class depends on the energy gain of protein folding, and the third on the association of the proteins to high-affinity binding sites in the IMS. PMID- 15817398 TI - When X-rays modify the protein structure: radiation damage at work. AB - The majority of 3D structures of macromolecules are currently determined by macromolecular crystallography, which employs the diffraction of X-rays on single crystals. However, during diffraction experiments, the X-rays can damage the protein crystals by ionization processes, especially when powerful X-ray sources at synchrotron facilities are used. This process of radiation damage generates photo-electrons that can get trapped in protein moieties. The 3D structure derived from such experiments can differ remarkably from the structure of the native molecule. Recently, the crystal structures of different oxidation states of horseradish peroxidase and nickel-containing superoxide dismutase were determined using crystallographic redox titration performed during the exposure of the crystals to the incident X-ray beam. Previous crystallographic analyses have not shown the distinct structures of the active sites associated with the redox state of the structural features of these enzymes. These new studies show that, for protein moieties that are susceptible to radiation damage and prone to reduction by photo-electrons, care is required in both the design of the diffraction experiment and the analysis and interpretation. PMID- 15817400 TI - Cutaneous lesions: benign and malignant. AB - This article reviews the clinical characteristics, histology, biologic behavior, and recommended treatment for several benign and malignant lesions that may arise on the head and neck. Nevus sebaceus and congenital melanocytic nevus are two benign lesions that can present at a size of several centimeters. Surgical excision may be considered for cosmetic purposes and to reduce the small risk for the development of malignancy within each lesion. Basal and squamous cell carcinoma, lentigo maligna and lentigo maligna melanoma, dermatofibrosarcoma protuberans, and Merkel cell carcinoma are malignant lesions for which surgical excision is the recommended treatment. Local flap reconstruction may be used to address the surgical defects resulting from excision of these benign and malignant conditions. PMID- 15817401 TI - Wound healing and physiology of skin flaps. AB - This article focuses on the dynamic process of wound healing, including the three phases of wound healing and the different types of wound healing. The physiology of skin flaps is described, with a focus on the vascular supply and biomechanics of skin flaps. The article reviews older and newer concepts of skin flap design. PMID- 15817402 TI - Minimally invasive options for cutaneous defects: secondary intention healing, partial closure, and skin grafts. AB - Several wound management options are available for defects of the head and neck, and choosing the best option requires consideration of several variables. The physical characteristics of the defect, the experience and preferences of the surgeon, and the desires and medicosocial situation of the patient may influence the final reconstructive decision. As the concepts and techniques in the field of reconstructive surgery advance, conservative wound management options should not be overlooked. This article reviews the minimally invasive options for the management of cutaneous defects, including second intention healing, partial closures, and skin grafts. The authors review the basic concepts of wound healing. PMID- 15817403 TI - Reconstruction of the auricle. AB - The intricate and delicate anatomy of the external ear (auricle) makes reconstruction a daunting task for beginning as well as experienced surgeons. The complex framework of the auricle is composed of graceful curves, peaks, and valleys of a thin pliable uniform thickness of cartilage. This article discusses techniques for reconstruction of the auricle. PMID- 15817404 TI - Soft tissue reconstruction of the forehead and temple. AB - Soft tissue reconstruction of the forehead and temple challenges facial plastic surgeons to balance esthetic goals with functional concerns. Understanding the muscular and neurovascular anatomy is essential to achieve these ends. Reconstructive considerations include maintaining hairlines and eyebrows, minimizing scarring, using relaxed skin tension lines, and preserving motor and sensory function. Reconstructive options range from healing by secondary intention to primary closure, skin grafts, local flaps, island flaps, or any combination of these techniques. PMID- 15817405 TI - Periorbital reconstruction: brows and eyelids. AB - The eyes and periorbital structures are principle components of the upper region of the face. A thorough understanding of orbital and periorbital anatomy and function is important for anyone wishing to perform reconstructive surgery in this region. This article discusses the unique challenges presented to the surgeon when performing reconstruction of the eyelids and periorbital structures. PMID- 15817406 TI - Current concepts in cheek reconstruction. AB - For cutaneous cheek defects, the reconstructive surgeon should strive to optimize tissue match and scar camouflage while minimizing the distortion to neighboring facial landmarks. The surgeon must have an intimate understanding of the local anatomy and must be able to use the appropriate flap for the given characteristics of each defect and patient. Knowledge of several types of flaps and versatility in modifying these flaps from their "textbook" geometry are required. This article highlights an approach to local flap reconstruction with a review of flap modification for particular situations. PMID- 15817407 TI - Repair of small nasal defects. AB - Nasal reconstruction is a challenging yet rewarding endeavor. Repair of nasal defects requires an appreciation of variations in nasal skin thickness and the influence of these differences on potential reconstructive methods. Multiple factors help determine the optimal method of repair, including the size of the defect relative to the amount of remaining skin, the depth and location of the defect, and the strength of the underlying nasal framework. Maintaining symmetry, contour, and function is essential for a successful nasal reconstruction. PMID- 15817408 TI - Lip reconstruction. AB - The lip presents a unique challenge to the facial plastic surgeon because of its prominent location, elegant form, and important functions. The anatomy and pathology relevant to the lip are considered herein, along with general concepts to guide lip surgery. The more common approaches to reconstructing lip defects are outlined, illustrated, and discussed. PMID- 15817409 TI - Tissue expansion in the head and neck. AB - Tissue expansion can be a valuable adjunct in reconstruction of large defects of the head and neck and is particularly useful in the scalp and forehead region. This article discusses the differences between conventional long-term expansion and rapid intraoperative expansion. Careful expander selection and application can allow extensive reconstructions with adjacent tissues that might otherwise be impossible. Complications are rare and usually avoidable with careful attention and technique. PMID- 15817410 TI - Scar revision. AB - Classic techniques of scar revision in general are ablative and remain the gold standard, although newer nonablative laser therapies, such as the frequency doubled neodymium:yttrium-aluminum-garnet laser, can help. Ablative therapies and techniques are examined. PMID- 15817411 TI - Complex nasal defects: structure and internal lining. AB - The development of nasal reconstruction concepts such as the three-layered (lining, structure, and covering) repair has led to remarkable esthetic and functional improvements. The authors present a variety of techniques for providing nasal structure and internal lining and discuss the application of these methods to specific nasal defects. PMID- 15817412 TI - Scalp reconstruction. AB - The unique properties of the human scalp affect the surgeon's ability to reconstruct defects in this area. The scalp has a rich vascular supply, the ability to withstand wounds of greater tension than other regions, and often has hair that can camouflage resulting scars or asymmetries. Nevertheless, most of the scalp is relatively inelastic and is ideally reconstructed with hair-bearing skin. These properties compromise the ease of effective reconstruction. However, multiple reconstructive options exist. The selection depends on the size and thickness of the defect, its location on the scalp, limiting comorbidities of the patient, and the need for immediate or delayed reconstruction. These options are reviewed in a systematic manner, organized by the factors outlined previously. PMID- 15817413 TI - The energetics of the chloroplast Tat protein transport pathway revisited. PMID- 15817414 TI - No need to shift the paradigm on the metabolic pathway to transitory starch in leaves. PMID- 15817416 TI - The ins and outs in membrane dynamics: tubulation and vesiculation. AB - Living cells constantly adjust the composition and size of their membrane systems to accommodate the demands for the housekeeping activities, to expand and reduce cell size, and to commit the cell for division. Although it is well known that vesicles are the vehicles to deliver and retrieve lipids and proteins to and from the membranes, the mechanisms allowing vesicles to pinch off from membranes or fuse into a flat lipid bilayer have been poorly understood, particularly in plants. Recent studies on dynamins and dynamin-related proteins in animals and plants now allow new concepts in membrane dynamics to be considered. PMID- 15817417 TI - Isoprenoids: an evolutionary pool for photoprotection. AB - Plants have evolved several mechanisms for getting rid of excess energy in photosynthetic membranes, some of which involve isoprenoid compounds. In all photosynthetic organisms, the carotenoids beta-carotene and zeaxanthin, and tocopherols serve an important photoprotective role, either by dissipating excess excitation energy as heat or by scavenging reactive oxygen species (ROS) and suppressing lipid peroxidation. Isoprene and some monoterpenes, diterpenes and other carotenoids also occur in some plant lineages. Compelling evidence indicates that these non-ubiquitous isoprenoids might be particularly relevant in adapting plants to adverse climatic conditions by serving as additional and/or alternative protection mechanisms. PMID- 15817418 TI - The PIN auxin efflux facilitators: evolutionary and functional perspectives. AB - It is widely believed that the PIN proteins are crucial for proper cellular coordination. Since the analysis of the Arabidopsis pin-formed mutant in 1991, and the subsequent cloning of AtPIN1, a further seven members of the family have been discovered. Here, we present an overview of this family of auxin efflux facilitators in monocot and dicot plants, summarizing their evolutionary history, expression profiles and, where appropriate, relating them to protein function. PMID- 15817419 TI - Related enzymes solve evolutionarily recurrent problems in the metabolism of carotenoids. AB - Chemical transformations of isoprenoids in plants and some bacteria and fungi lead to the production of various conjugated products, including carotenoids. Carotenoids can be cleaved to generate apocarotenoid precursors for signaling molecules such as abscisic and retinoic acids, and for the photosensory pigment retinal. The enzymes that catalyze the various transformations of carotenoids and apocarotenoids are closely related. This evolutionarily distant conservation is unexpected and intriguing. Many aspects of the metabolism of retinoids in vertebrates remain controversial and poorly understood. Because few chemical reactions are possible for this group of compounds, furthering our knowledge of isoprenoid transformation in plants could be beneficial to our understanding of how retinoids and carotenoids are transformed in vertebrates. PMID- 15817420 TI - Oxidative tailoring of carotenoids: a prospect towards novel functions in plants. AB - Carotenoids not only play a crucial role in their intact form but also are an important reservoir of lipid-derived bioactive mediators. The process is initiated by tailoring enzymes that cleave carotenoids into apocarotenoids. Apocarotenoids act as visual or volatile signals to attract pollinating and seed dispersal agents, and are also key players in allelopathic interactions and plant defense. Recent studies show that the loss of these cleavage enzymes induces the development of axillary branches, indicating that apocarotenoids convey signals that regulate plant architecture. Here, we describe these molecules and the current understanding of their biosynthesis and functions. PMID- 15817421 TI - New insights into nitric oxide metabolism and regulatory functions. AB - Nitric oxide (NO) has been intensively studied to elucidate the role of this enigmatic signaling molecule in plant development, metabolism and disease responses. Many studies using pharmacological and biochemical tools have demonstrated that NO functions in hormone responses, programmed cell death, defense gene induction and signal transduction pathways. NO originates from two sources in plants: nitrite and arginine. Recent studies using mutants and transgenic plants have confirmed these key findings and have gone further to identify (i) a new mechanism to modulate NO bioactivity involving hemoglobin, (ii) a gene involved in arginine-dependent NO synthesis, and (iii) a novel function for NO signaling in flowering. These findings continue to elucidate the expanding role of NO in plant biology. PMID- 15817422 TI - Image fusion of CT and MRI for the visualization of the auditory and vestibular system. AB - The aim of this study was to perform a realistic visualization of the auditory and vestibular system using volume data sets from high-resolution computed tomography (HR-CT) and high-resolution magnetic resonance imaging (HR-MRI). - In 10 patients with conductive and/or sensorineural hearing loss, vertigo and tinnitus, HR-CT and HR-MRI of the petrous bone were performed consecutively. CT was performed with a 16-slice computed tomography scanner using a high spatial resolution. MRI was performed with a 3.0 Tesla scanner using a three-dimensional constructive interference in steady state (3D-CISS) gradient-echo, and T2 weighted, unenhanced and gadolinium (GD)-enhanced T1-weighted turbo spin-echo sequences. The middle ear structures were interactively segmented and visualized with a color-coded shaded-surface rendering method using the HR-CT volume data sets. The inner ear structures were interactively segmented and visualized with a color-coded shaded-surface rendering method using the high-resolution 3D-CISS MRI volume data sets. Finally, both shaded-surface rendered models were superimposed semi-automatically using a commercial available software program to visualize the auditory and vestibular system. - The representation of the middle and inner ear structures with image fusion of HR-CT and HR-MRI takes advantage of both the high bony contrast of HR-CT and the high soft tissue contrast discrimination and sensitivity to fluids of HR-MRI, as well as the high spatial resolution of both modalities. In comparison to the fused axial CT/MRI, the images of 3D CT/MRI fusion facilitates a clear representation and better spatial orientation. - The middle and inner ear consists of bony structures, soft tissue structures and fluid-filled spaces. For this reason, the image fusion of volume data sets from HR-CT and HR-MRI allowed an optimized and realistic visualization of the auditory and vestibular system. PMID- 15817423 TI - Long-term consequences of treatment interruptions in chronically HIV-1-infected patients. AB - OBJECTIVE: To evaluate the long-term effects of antiretroviral treatment (ART) interruptions on metabolic, immunological, virological and clinical outcomes in chronically HIV-1 infected patients. METHODS: Multi-centric, prospective, controlled 24-month cohort study in HIV-1 infected patients interrupting ART once or several times and for at least two weeks. Patients were compared to a frequency-matched control group continuing on ART. RESULTS: A total of 399 HIV-1 infected patients were included, among them 133 patients with treatment interruption (TI) and 266 control patients. Baseline characteristics were well matched. Median baseline CD4 cell count was 379/microl in TI-patients and 410/microl in control patients (p = ns). Median duration of the first TI was 1.1 months, and 37 % of patients had two or further TI's. Whereas CD4 cell count in control patients had increased significantly by a median of 67/microl at month 24 (p<0.0001), median CD4 cell count at month 24 in the TI-patients did not differ significantly from baseline. However, two-year AIDS-free survival was not significantly different between TI- and control patients. Liver enzymes and blood lipids improved significantly during TI. CONCLUSION: TI was associated with a significant immunological disadvantage at 24-month follow-up compared to continued ART. In this relatively immunocompetent cohort, however, TI's did not lead to an increased risk of disease progression within two years of follow-up. PMID- 15817424 TI - Cutaneous mucinosis and skin necrosis complicates interferon alfacon-1 (consensus interferon) treatment of chronic hepatitis C. AB - A 59-year-old Caucasian suffering from chronic hepatitis C started daily subcutaneous self-injections of interferon alfacon-1 (consensus interferon) according to the protocol of a randomised multicentre study. At week 10, he developed painful erythematous lesions at two injection sites. Consensus interferon dosage was reduced, and eventually, the lesions healed with small areas of scarring due to central necrosis. At week 51, again large areas of erythematous thickened skin arose at two other injection sites followed by substantial ulceration and central necrosis. Skin biopsy revealed excessive dermal mucin deposition so that cutaneous mucinosis was diagnosed. After 3-6 months, the lesions healed with central scarring. Due to a persistently negative HCV-PCR from serum (from week 12 onwards), consensus interferon treatment was not stopped but continued until week 60 according to the study protocol. - Cutaneous mucinosis has not been previously reported to complicate treatment with consensus interferon in patients with chronic hepatitis C and should therefore be added to the list of dermatological side-effects associated with interferon alfacon-1 therapy. PMID- 15817425 TI - Oral silymarin for chronic hepatitis C - a retrospective analysis comparing three dose regimens. AB - To investigate the effects of silymarin on aminotransferase levels in patients with chronic hepatitis C, a standardized treatment with 420mg, 840mg or 1260mg per day was performed in patients of our clinic, who were not eligible for treatment with pegylated interferon and ribavirin. Aminotransferase levels were determined before, at 3-6 week intervals during and at the end of treatment. Predefined inclusion criteria for the retrospective analysis were persistently elevated alanine aminotransferase (ALT) levels (at least 6 months prior to and at beginning of the treatment) and treatment duration of at least three weeks. Liver cirrhosis CHILD B or C, interferon therapy within the last three months before treatment with silymarin, alcohol use >30 g/d, coinfection with hepatitis B virus or other severe diseases were exclusion criteria. According to these criteria 40 patients (13 with 420mg, 20 with 840mg and 7 with 1260mg silymarin per day) were eligible for the analysis. The mean treatment period was 125 +/- 78 days. ALT, aspartate aminotransferase and gamma glutamyltransferase levels did not change significantly from baseline in any group and there were no differences between the treatment groups. Bilirubin and prothrombine time were normal in all but one patient and remained unchanged. Silymarin therapy had no side effects. Silymarin at the doses used, does not improve elevated aminotransferases in patients with chronic hepatitis C. PMID- 15817426 TI - Epidemiological and health economical evaluation of intraoperative antibiosis as a protective agent against endophthalmitis after cataract surgery. AB - OBJECTIVE: To evaluate risk factors for endophthalmitis after cataract surgery and to retest recent findings on the protective effect of intraoperative antibiosis and the promoting effect of the clear corneal as compared to sclerocorneal incision. In addition, the economics of intraocular intraoperative antibiosis as a prophylaxis in cataract surgery are asketched. DESIGN: Survey study. PARTICIPANTS: Five hundred thirty-eight ophthalmosurgical centres in Germany. MAIN OUTCOME MEASURE: epidemiological evaluation: responder specific endophthalmitis incidence in year 2000; economical evaluation: direct cost analysis based on incidence data and local cost estimates (health service's perspective). RESULTS: A total of 310 (58%) questionnaires were computed resulting in an overall count of 404,356 cataract surgeries and 291 self-reported endophthalmitis cases (crude rate 0.072%). The risk of postoperative endophthalmitis for sclerocorneal versus clear corneal incisions was not significantly reduced (relative risk 0.97, 99% confidence interval 0.69-1.38). The hypothesis of a protective effect of intraocular antibiosis could be confirmed by a significantly decreased risk ratio of 0.69 (99% confidence interval 0.48-0.99) indicating a significant benefit from intraoperative intraocular antibiosis. A similar tendency was observed for an intraoperative periocular antibiosis with a significantly reduced risk ratio of 0.68 (99% confidence interval 0.49-0.96). These risk estimates had been adjusted for the size of the surgical centre: a significantly reduced risk ratio of 0.70 (99% confidence interval 0.49-0.98) for postoperative endophthalmitis was observed for local centres. Cost evaluation for the prophylactic use of intraocular intraoperative antibiosis in cataract surgery revealed an economically relevant decrease in direct endophthalmitis associated costs. CONCLUSIONS: Whereas this 2000 appraisal of a recent survey in 1996 could not reproduce the benefit of sclerocorneal incision, the protective effect of intraoperative intraocular antibiotic prophylaxis could be confirmed. However, the results of this survey have to be interpreted with care, since it is not based on individual case information, but rather on aggregate questionnaire data. PMID- 15817427 TI - Effects of endotoxin on serum chemokines in man. AB - AIM: Endotoxin is known to be a primary initiator of sepsis and septic shock. Migration of immunocompetent cells due to chemotactic attraction plays a central role in the initiation of the immune response. Two major groups of chemokines can be distinguished: C-x-C chemokines like Interleukin-8 attract mainly neutrophils, C-C chemokines (e.g. RANTES) attract monocytes and T-cells. The aim of this study was to get further insight into chemokine profiles after a single endotoxin bolus in man. MATERIALS AND METHODS: We investigated the effect of systemically administered endotoxin (4ng/kg BW i.v.) in 8 healthy volunteers. Clinical data (heart rate, mean arterial pressure, temperature), serum levels of IL-8, and RANTES, as well as white blood cell count were obtained before and hourly for five hours after endotoxin administration. RESULTS: Heart rate and MAP showed significant changes (p<0.05) after 2-3 hours. All volunteers presented with low grade fever after 2 hours. WBC was elevated 43% and 63% after 4 and 5 hours, respectively. Both chemokines were significantly different from baseline two hours after endotoxin challenge: While IL-8 was significantly increased RANTES serum levels were diminished. CONCLUSION: From our data we conclude that this endotoxin model was effective to mimic the clinical appearance of sepsis. Chemokines like IL-8 and RANTES are integrated in the early immune response to endotoxin challenge in man. PMID- 15817428 TI - Hemofiltration does not influence early S-100B serum levels in septic shock patients receiving stress doses of hydrocortisone or placebo. AB - BACKGROUND: The prognosis in patients with hyperdynamic septic shock correlates with the presence and the severity of septic encephalopathy. However, the neurological evaluation is considerably influenced by the use of analgesia sedation during mechanical ventilation. An early concentration peak of the neuroprotein S-100B in serum reflects both cellular damage at an increased permeability of the blood-brain-barrier and a delayed renal elimination. Thus, the objective of this study was to analyze the effect of continuous veno-venous hemofiltration (CVVH) on early S-100B serum levels in septic shock patients, who were treated with either stress doses of hydrocortisone or placebo. METHODS: Twenty-four consecutive patients, who met the ACCP / SCCM criteria for septic shock, were enrolled in this prospective, randomised, double-blind, single-center trial. The severity of illness at recruitment was graded using the APACHE II and SAPS II scoring systems; the MODS was described by the SOFA score. All patients were prospectively randomised to receive either stress doses of hydrocortisone or placebo. Hydrocortisone was started in 12 patients with a loading dose of 100 mg and followed by a continuous infusion of 0.18 mg/kg/h for 6 days. RESULTS: Median S-100B serum levels of the hydrocortisone group decreased from 0.32 ng/ml (0.19 3.60) at study entry to 0.07 ng/ml (0.04 - 0.32) 6 days later without significant differences compared to the placebo group. Patients undergoing CVVH showed significantly higher S-100B serum values compared to patients without CVVH (p<0.001). However, initial median S-100B serum levels of the CVVH group even increased from 0.92 ng/ml (0.16 - 4.63) to 2.33 ng/ml (0.59 - 2.44) 30 hours after study entry, reaching data ranges already known in patients with out-of hospital cardiac arrest or severe traumatic brain injury. CONCLUSION: Early S 100B serum levels in septic shock patients receiving either stress doses of hydrocortisone or placebo were not influenced by CVVH. For the first time, we observed a similar extent of S-100B serum increase in CVVH patients, who had significantly higher S-100B serum values compared to those without CVVH, as reported for out-of-hospital cardiac arrest or severe traumatic brain injury. Hypercortisolemia induced by the infusion of stress doses of hydrocortisone did not significantly reduce early S-100B serum concentrations with time. PMID- 15817429 TI - Staging laparoscopy in gastric cancer. AB - BACKGROUND: As patients with gastric cancer are offered choices between surgical resection, neoadjuvant or palliative chemotherapy, or symptomatic relief alone, the need for accurate preoperative staging becomes apparent. Laparoscopy has been suggested as an appropriate staging modality in a variety of upper gastrointestinal malignancies. METHODOLOGY: Staging laparoscopy was performed in 45 patients with potentially resectable gastric cancer as determined by physical examination, gastroscopy, endosonography, transcutaneous ultrasonography and current generation computed tomography. Conventional clinic staging and laparoscopic staging according to the Tumor-Node-Metastases classification of the International Union against Cancer were registered separately on a database. Results were then compared to evaluate the agreement of both staging procedures. RESULTS: Compared to conventional staging, laparoscopy resulted in up staging of 23 patients (51.1%). In 10 patients of them peritoneal seeding was first found during laparoscopy, whereas additional liver metastases were detected in 3 patients and Krukenberg's tumor in one. As a consequence, the therapy planning was changed and laparotomy was avoided in 14 of these patients as the first operative procedure. Sensitivity of clinical staging was especially poor for stage IV tumors (5.3%) and for the majority of stage IIIB tumors (42.9%). Cytologic examination of peritoneal fluid had no additional information in our series. CONCLUSIONS: The value of laparoscopy in staging patients with gastric carcinoma could be demonstrated in this study. It is a safe and effective staging modality, helping to avoid unnecessary laparotomies and providing new means of directing appropriate treatment strategy. PMID- 15817430 TI - Directionally compliant legs influence the intrinsic pitch behaviour of a trotting quadruped. AB - Limb design is well conserved among quadrupeds, notably, the knees point forward (i.e. cranial inclination of femora) and the elbows point back (i.e. caudal inclination of humeri). This study was undertaken to examine the effects of joint orientation on individual leg forces and centre of mass dynamics. Steady-speed trotting was simulated in two quadrupedal models. Model I had the knee and elbow orientation of a quadruped and model II had a reversed leg configuration in which knees point back and elbows point forward. The model's legs showed directional compliance determined by the orientation of the knee/elbow. In both models, forward pointing knees/elbows produced a propulsive force bias, while rearward pointing knees/elbows produced a braking force bias. Hence, model I showed the same pattern of hind-leg propulsion and fore-leg braking observed in trotting animals. Simulations revealed minimal pitch oscillations during steady-speed trotting of model I, but substantially greater and more irregular pitch oscillations of model II. The reduced pitch oscillation of model I was a result of fore-leg and hind-leg forces that reduced pitching moments during early and late stance, respectively. This passive mechanism for reducing pitch oscillations was an emergent property of directionally compliant legs with the fore-hind configuration of model I. Such intrinsic stability resulting from mechanical design can simplify control tasks and lead to more robust running machines. PMID- 15817431 TI - Ecological speciation in tropical reef fishes. AB - The high biodiversity in tropical seas provides a long-standing challenge to allopatric speciation models. Physical barriers are few in the ocean and larval dispersal is often extensive, a combination that should reduce opportunities for speciation. Yet coral reefs are among the most species-rich habitats in the world, indicating evolutionary processes beyond conventional allopatry. In a survey of mtDNA sequences of five congeneric west Atlantic reef fishes (wrasses, genus Halichoeres) with similar dispersal potential, we observed phylogeographical patterns that contradict expectations of geographical isolation, and instead indicate a role for ecological speciation. In Halichoeres bivittatus and the species pair Halichoeres radiatus/brasiliensis, we observed strong partitions (3.4% and 2.3% divergence, respectively) between adjacent and ecologically distinct habitats, but high genetic connectivity between similar habitats separated by thousands of kilometres. This habitat partitioning is maintained even at a local scale where H. bivittatus lineages are segregated between cold- and warm-water habitats in both Bermuda and Florida. The concordance of evolutionary partitions with habitat types, rather than conventional biogeographical barriers, indicates parapatric ecological speciation, in which adaptation to alternative environmental conditions in adjacent locations overwhelms the homogenizing effect of dispersal. This mechanism can explain the long-standing enigma of high biodiversity in coral reef faunas. PMID- 15817432 TI - Muscle directly meets the vast power demands in agile lizards. AB - Level locomotion in small, agile lizards is characterized by intermittent bursts of fast running. These require very large accelerations, often reaching several times g. The power input required to increase kinetic energy is calculated to be as high as 214 W kg(-1) muscle (+/-20 W kg(-1) s.e.; averaged over the complete locomotor cycle) and 952 W kg(-1) muscle (+/-89 W kg(-1) s.e.; instantaneous peak power). In vitro muscle experiments prove that these exceptional power requirements can be met directly by the lizard's muscle fibres alone; there is no need for mechanical power amplifying mechanisms. PMID- 15817433 TI - A simple model for the within-host dynamics of a protozoan parasite. AB - The dynamics of parasite-host systems can be complicated if the parasite life cycle contains an obligatory environmental stage and if the hosts' immunity increases upon re-infection. The dynamics then greatly depend on the relation between infection history and parasite uptake and excretion of individual hosts. In an effort to better understand such systems, we study Eimeria spp. in chickens as our model. In this paper we take a first step and study the within-host dynamics of Eimeria spp. transmitted through oocysts in the environment, with a mathematical model for the parasite life cycle in discrete time, interacting with a single variable describing the immune response. The model can explain various types of oocyst input-output behaviour as described in previous experiments, in particular the characteristic crowding effect, which causes a decreasing oocyst production with increasing single dose oocyst uptake. Oocyst excretion during constant oocyst uptake (trickle infection) and the immunizing effect of single and trickle infections also appears in accordance with published experiments. The model seems a good description of oocyst input-output behaviour in individual hosts; it provides a solid basis for the study of between-host dynamics, where individuals interact in a common environment, thereby affecting their own and each other's infection pattern. PMID- 15817434 TI - Compensatory response 'defends' energy levels but not growth trajectories in brown trout, Salmo trutta L. AB - Compensatory growth is an organism's reaction to buffer deviations from targeted trajectories. We explored the compensatory patterns of juvenile brown trout under field and laboratory conditions. Divergence of size and condition trajectories was induced by manipulating food levels in the laboratory and then releasing the trout into a river. In the stream, the length trajectories of food-restricted and control fish were parallel, but food-restricted fish exhibited partial compensation for mass and rapid recovery of condition. A laboratory experiment on similar sized fish did not provide evidence for compensatory growth in length or mass. In contrast, data matched the compensatory patterns shown in the stream: length trajectories were parallel and the convergence of mass trajectories ceased as soon as food-restricted fish recovered condition to the level of controls. These results show that (i) brown trout did not compensate for depression in structural growth and (ii) mass recovery was targeted to reinstate condition or energy reserves, but not size at a given age. This does not support the common view that compensatory growth can be a general response to growth depression. Rather, compensation in other salmonids could be related to size thresholds associated with developmental switches at the onset of sexual maturation and migration. PMID- 15817435 TI - Morphological evolution, ecological diversification and climate change in rodents. AB - Among rodents, the lineage from Progonomys hispanicus to Stephanomys documents a case of increasing size and dental specialization during an approximately 9 Myr time-interval. On the contrary, some contemporaneous generalist lineages like Apodemus show a limited morphological evolution. Dental shape can be related to diet and can be used to assess the ecological changes along the lineages. Consequently, size and shape of the first upper molar were measured in order to quantify the patterns of morphological evolution along both lineages and compare them to environmental trends. Climatic changes do not have a direct influence on evolution, but they open new ecological opportunities by changing vegetation and allow the evolution of a specialist like Stephanomys. On the other hand, environmental changes are not dramatic enough to destroy the habitat of a long term generalist like Apodemus. Hence, our results exemplify a case of an influence of climate on the evolution of specialist species, although a generalist species may persist without change. PMID- 15817436 TI - Bite club: comparative bite force in big biting mammals and the prediction of predatory behaviour in fossil taxa. AB - We provide the first predictions of bite force (BS) in a wide sample of living and fossil mammalian predators. To compare between taxa, we calculated an estimated bite force quotient (BFQ) as the residual of BS regressed on body mass. Estimated BS adjusted for body mass was higher for marsupials than placentals and the Tasmanian devil (Sarcophilus harrisii) had the highest relative BS among extant taxa. The highest overall BS was in two extinct marsupial lions. BFQ in hyaenas were similar to those of related, non-osteophagous taxa challenging the common assumption that osteophagy necessitates extreme jaw muscle forces. High BFQ in living carnivores was associated with greater maximal prey size and hypercarnivory. For fossil taxa anatomically similar to living relatives, BFQ can be directly compared, and high values in the dire wolf (Canis dirus) and thylacine (Thylacinus cynocephalus) suggest that they took relatively large prey. Direct inference may not be appropriate where morphologies depart widely from biomechanical models evident in living predators and must be considered together with evidence from other morphological indicators. Relatively low BFQ values in two extinct carnivores with morphologies not represented among extant species, the sabrecat, Smilodon fatalis, and marsupial sabretooth, Thylacosmilus atrox, support arguments that their killing techniques also differed from extant species and are consistent with 'canine-shear bite' and 'stabbing' models, respectively. Extremely high BFQ in the marsupial lion, Thylacoleo carnifex, indicates that it filled a large-prey hunting niche. PMID- 15817437 TI - Mineralization of soft-part anatomy and invading microbes in the horseshoe crab Mesolimulus from the Upper Jurassic Lagerstatte of Nusplingen, Germany. AB - A remarkable specimen of Mesolimulus from the Upper Jurassic (Kimmeridgian) of Nusplingen, Germany, preserves the musculature of the prosoma and associated microbes in three dimensions in calcium phosphate (apatite). The musculature of Mesolimulus conforms closely to that of modern horseshoe crabs. Associated with the muscles are patches of mineralized biofilm with spiral and coccoid forms. This discovery emphasizes the potential of soft-bodied fossils as a source for increasing our knowledge of the diversity of fossil microbes in particular settings. PMID- 15817438 TI - The distributions of the duplicate oestrogen receptors ER-beta a and ER-beta b in the forebrain of the Atlantic croaker (Micropogonias undulatus): evidence for subfunctionalization after gene duplication. AB - Teleost fishes have three distinct oestrogen receptor (ER) subtypes: ER-alpha, ER beta a (or ER-gamma) and ER-beta b. ER-beta a and ER-beta b arose from a duplication of an ancestral ER-beta gene early in the teleost lineage. Here, we describe the distribution of the three ER mRNAs in the hypothalamus and cerebellum of the Atlantic croaker to address two issues: the specific functions of multiple ERs in the neuroendocrine system and the evolution and fate of duplicated genes. ER-alpha was detected in nuclei of the preoptic area (POA) and hypothalamus previously shown to possess ER-alphas in teleosts. AcER-beta b, but not ER-beta a, labelling was detected in the magnocellular neurons of the POA, nucleus posterior tuberis, the nucleus recessus posterior and cerebellum. By contrast, acER-beta a, but not ER-beta b, was detected in the dorsal anterior parvocellular POA and suprachiasmatic nucleus. Both ER-betas were found in posterior parvocellular and ventral anterior POA nuclei, the ventral hypothalamus, and periventricular dorsal hypothalamus. The differences we observed in ER subtype mRNA distribution within well-characterized brain nuclei suggest that ER-beta a and ER-beta b have distinct functions in the neuroendocrine control of reproduction and behaviour, and provide evidence that the teleost ER-beta paralogues have partitioned functions of the ancestral ER beta gene they shared with tetrapods. PMID- 15817439 TI - Cadherins in maternal-foetal interactions: red queen with a green beard? AB - Cadherins are homophilic cell surface adhesion proteins, some of which mediate interactions between maternal and foetal tissues during mammalian pregnancy. David Haig suggested that these proteins may exhibit 'green-beard gene' effects, whereby the nature of binding between identical alleles in mother and foetus leads to differential levels of resource transfer. The selfish effects of such self-recognizing alleles should, however, be suppressed over evolutionary time by unlinked genes, which is expected to lead to antagonistic coevolution between placentally expressed cadherins and unlinked modifiers. Such molecular coevolution should leave a signature of positive selection, with high ratios of non-synonymous to synonymous amino acid substitution. We present evidence that three placentally expressed cadherin genes, E-cadherin, P-cadherin and VE cadherin, have been subject to positive selection. By contrast, a 'control' cadherin that is not expressed in the placenta, H-cadherin, showed no evidence of selection. These results provide support for the hypothesis that the cadherin genes involved in maternal-foetal interactions have been subject to green-beard effect mutations over the course of evolutionary history, leading to antagonistic coevolution with suppressing elements from the parliament of genes. PMID- 15817440 TI - Ambient noise and the design of begging signals. AB - The apparent extravagance of begging displays is usually attributed to selection for features, such as loud calls, that make the signal costly and hence reliable. An alternative explanation, however, is that these design features are needed for effective signal transmission and reception. Here, we test the latter hypothesis by examining how the begging calls of tree swallow (Tachycineta bicolor) nestlings and the response to these calls by parents are affected by ambient noise. In a field study, we found that call length, amplitude and frequency range all increased with increasing noise levels at nests. In the laboratory, however, only call amplitude increased in response to the playback of noise to nestlings. In field playbacks to parents, similar levels of noise abolished parental preferences for higher call rates, but the preference was restored when call amplitude was increased to the level that nestlings had used in the laboratory study. Our results show that nestling birds, like other acoustic signallers, consistently increase call amplitude in response to ambient noise and this response appears to enhance discrimination by receivers. Thus, selection for signal efficacy may explain some of the seemingly extravagant features of begging displays. PMID- 15817441 TI - Genetic variation for an aphid wing polyphenism is genetically linked to a naturally occurring wing polymorphism. AB - Many polyphenisms are examples of adaptive phenotypic plasticity where a single genotype produces distinct phenotypes in response to environmental cues. Such alternative phenotypes occur as winged and wingless parthenogenetic females in the pea aphid (Acyrthosiphon pisum). However, the proportion of winged females produced in response to a given environmental cue varies between clonal genotypes. Winged and wingless phenotypes also occur in males of the sexual generation. In contrast to parthenogenetic females, wing production in males is environmentally insensitive and controlled by the sex-linked, biallelic locus, aphicarus (api). Hence, environmental or genetic cues induce development of winged and wingless phenotypes at different stages of the pea aphid life cycle. We have tested whether allelic variation at the api locus explains genetic variation in the propensity to produce winged females. We assayed clones from an F2 cross that were heterozygous or homozygous for alternative api alleles for their propensity to produce winged offspring. We found that clones with different api genotypes differed in their propensity to produce winged offspring. The results indicate genetic linkage of factors controlling the female wing polyphenism and male wing polymorphism. This finding is consistent with the hypothesis that genotype by environment interaction at the api locus explains genetic variation in the environmentally cued wing polyphenism. PMID- 15817442 TI - Background-matching and disruptive coloration, and the evolution of cryptic coloration. AB - Cryptic prey coloration typically bears a resemblance to the habitat the prey uses. It has been suggested that coloration which visually matches a random sample of the background maximizes background matching. We studied this previously untested hypothesis, as well as another, little studied principle of concealment, disruptive coloration, and whether it could, acting in addition to background matching, provide another plausible means of achieving camouflage. We presented great tits (Parus major) with artificial background-matching and disruptive prey (DP), and measured detection times. First, we studied whether any random sample of a background produces equally good crypsis. This turned out to not be the case. Next, we compared the DP and the best background-matching prey and found that they were equally cryptic. We repeated the tests using prey with all the coloration elements being whole, instead of some of them being broken by the prey outline, but this did not change the result. We conclude that resemblance of the background is an important aspect of concealment, but that coloration matching a random visual sample of the background is neither sufficient nor necessary to minimize the probability of detection. Further, our study lends empirical support to the principle of disruptive coloration. PMID- 15817444 TI - Acute paralysis following "a bad potato": a case of botulism. AB - PURPOSE: Intensivists often encounter patients with respiratory failure as a result of neuromuscular disease, however, acute neuro-muscular syndromes are less common. We present a case of food borne Clostridium botulism and discuss the diagnostic and therapeutic considerations. CLINICAL FINDINGS: A 35-yr-old healthy male presented with abdominal pain and blurred vision 12 hr after ingesting a "bad" potato. During the next 17 hr, the patient demonstrated a gradual descending paralysis which ultimately resulted in no cranial nerve function and 0/5 strength in all extremities. Sensation was intact. The patient required intubation and mechanical ventilation. His blood count, biochemical profile, computerized tomography and magnetic resonance imaging of the head were normal. A lumbar puncture revealed no abnormalities. Due to the rapid deterioration and presentation of 'descending' paralysis, botulism was suspected. The patient was treated empirically with botulinum anti-toxin. Samples of blood, stool and gastric contents were cultured for the presence of Clostridium botulinum and its toxin and these tests were positive for botulinum toxin A 12 days later. The patient's neuromuscular function gradually improved over a prolonged period of time. Six and one-half months after his initial presentation, the patient was discharged home after completing an aggressive rehabilitation program. CONCLUSIONS: Botulism is a rare syndrome and presents as an acute, afebrile, descending paralysis beginning with the cranial nerves. If suspected, botulinum anti-toxin should be considered, particularly within the first 24 hr of onset of symptoms. Confirmation of the presence of botulinum requires days therefore the diagnosis and management rely on history and physical examination. PMID- 15817443 TI - Bone marrow clonogenic capability, cytokine production, and thymic output in patients with common variable immunodeficiency. AB - In patients with primary Ab deficiencies, hematological and immunological abnormalities are frequently observed. A regenerative failure of hemopoietic stem/progenitor cells has been hypothesized. We evaluated in the bone marrow (BM) of 11 patients with common variable immunodeficiency, the phenotype of BM progenitors and their in vitro growth by colony-forming cell (CFC) and long-term culture (LTC) assays. A significant decrease in erythroid and mixed CFC and, to a greater extent, in primitive LTC-CFC progenitors was observed in patients compared with healthy controls. The frequency of BM pre-B and pro-B cells correlated directly with the absolute number of CD19+ lymphocytes. BM cells cultured in vitro produced spontaneously lower amounts of IL-2 and elevated levels of TNF-alpha compared with controls, indicating a skewing toward a proapoptotic cytokine pattern. In addition, stromal cells generated after BM LTC secreted less IL-7 and displayed by immunohistochemistry an altered phenotype. These findings were associated with a significant decrease in naive Th cells coexpressing CD31 in the peripheral blood. These results indicate an impaired growth and differentiation capacity of progenitor cells in patients with common variable immunodeficiency. PMID- 15817445 TI - Percutaneous transhepatic biliary dilatation under thoracic epidural analgesia in a patient with a recent myocardial infarction. PMID- 15817446 TI - Artificially lipid-anchored proteins can elicit clustering-induced intracellular signaling events in Jurkat T-lymphocytes independent of lipid raft association. AB - We have incorporated artificial lipid-anchored streptavidin conjugates with fully saturated or polyunsaturated lipid anchors into the plasma membranes of Jurkat T lymphocytes to assess previous conclusions that the activation of signaling processes induced in these cells by clustering of endogenous glycosylphosphatidylinositol-anchored proteins or ganglioside GM1 depends specifically on the association of these membrane components with lipid rafts. Lipid-anchored streptavidin conjugates could be incorporated into Jurkat or other mammalian cell surfaces by inserting biotinylated phosphatidylethanolamine polyethyleneglycols (PE-PEGs) and subsequently binding streptavidin to the cell incorporated PE-PEGs. Saturated dipalmitoyl-PE-PEG-streptavidin conjugates prepared in this manner partitioned substantially into the detergent-insoluble membrane fraction isolated from Jurkat or fibroblast cells, whereas polyunsaturated dilinoleoyl-PE-PEG-anchored conjugates were wholly excluded from this fraction, consistent with the differences in the affinities of the two types of lipid anchors for liquid-ordered membrane domains. Remarkably, however, antibody-mediated cross-linking of either dipalmitoyl- or dilinoleoyl-PE-PEG anchored streptavidin conjugates in Jurkat cells induced elevation of cytoplasmic calcium levels and tyrosine phosphorylation of the scaf-folding protein linker of T-cell activation in a manner similar to that observed upon cross-linking of endogenous CD59 or ganglioside GM1. The amplitude of the cross-linking-stimulated elevation of cytoplasmic calcium moreover showed an essentially identical dependence on the level of incorporated streptavidin conjugate for either type of lipid anchor. Confocal fluorescence microscopy revealed that PE-PEG-streptavidin conjugates with saturated versus polyunsaturated anchors showed very similar surface distributions vis a vis GM1 or CD59 under conditions where one or both species were cross-linked. These results indicate that cross-linking of diverse proteins anchored only to the outer leaflet of the plasma membrane can induce activation of Jurkat T-cell-signaling responses, but they appear to contradict previous suggestions that this phenomenon rests specifically on the association of such species with lipid rafts. PMID- 15817447 TI - Thrombin modulates the expression of a set of genes including thrombospondin-1 in human microvascular endothelial cells. AB - Thrombospondin-1 (THBS1) is a large extracellular matrix glycoprotein that affects vasculature systems such as platelet activation, angiogenesis, and wound healing. Increases in THBS1 expression have been liked to disease states including tumor progression, atherosclerosis, and arthritis. The present study focuses on the effects of thrombin activation of the G-protein-coupled, protease activated receptor-1 (PAR-1) on THBS1 gene expression in the microvascular endothelium. Thrombin-induced changes in gene expression were characterized by microarray analysis of approximately 11,000 different human genes in human microvascular endothelial cells (HMEC-1). Thrombin induced the expression of a set of at least 65 genes including THBS1. Changes in THBS1 mRNA correlated with an increase in the extracellular THBS1 protein concentration. The PAR-1-specific agonist peptide (TFLLRNK-PDK) mimicked thrombin stimulation of THBS1 expression, suggesting that thrombin signaling is through PAR-1. Further studies showed THBS1 expression was sensitive to pertussis toxin and protein kinase C inhibition indicating G(i/o)- and G(q)-mediated pathways. THBS1 up-regulation was also confirmed in human umbilical vein endothelial cells stimulated with thrombin. Analysis of the promoter region of THBS1 and other genes of similar expression profile identified from the microarray predicted an EBOX/EGRF transcription model. Expression of members of each family, MYC and EGR1, respectively, correlated with THBS1 expression. These results suggest thrombin formed at sites of vascular injury increases THBS1 expression into the extracellular matrix via activation of a PAR-1, G(i/o), G(q), EBOX/EGRF-signaling cascade, elucidating regulatory points that may play a role in increased THBS1 expression in disease states. PMID- 15817448 TI - Dissecting the structural determinants of the stability of cholesterol oxidase containing covalently bound flavin. AB - Cholesterol oxidase from Brevibacterium sterolicum is a monomeric flavoenzyme catalyzing the oxidation and isomerization of cholesterol to cholest-4-en-3-one. This protein is a class II cholesterol oxidases, with the FAD cofactor covalently linked to the enzyme through the His(69) residue. In this work, unfolding of wild type cholesterol oxidase was compared with that of a H69A mutant, which does not covalently bind the flavin cofactor. The two protein forms do not show significant differences in their overall topology, but the urea-induced unfolding of the H69A mutant occurred at significant lower urea concentrations than wild type (approximately 3 versus approximately 5 M, respectively), and the mutant protein had a melting temperature approximately 10-15 degrees C lower than wild type in thermal denaturation experiments. The different sensitivity of the various spectroscopic features used to monitor protein unfolding indicated that in both proteins a two-step (three-state) process occurs. The presence of an intermediate was more evident for the H69A mutant at 2 m urea, where catalytic activity and tertiary structure were lost, and new hydrophobic patches were exposed on the protein surface, resulting in protein aggregation. Comparative analysis of the changes occurring upon urea and thermal treatment of the wild type and H69A protein showed a good correlation between protein instability and the elimination of the covalent link between the flavin and the protein. This covalent bond represents a structural device to modify the flavin redox potentials and stabilize the tertiary structure of cholesterol oxidase, thus pointing to a specific meaning of the flavin binding mode in enzymes that carry out the same reaction in pathogenic versus non-pathogenic bacteria. PMID- 15817449 TI - A role for the protease-sensitive loop region of Shiga-like toxin 1 in the retrotranslocation of its A1 domain from the endoplasmic reticulum lumen. AB - Shiga-like toxin I (Slt-I) is a ribosome-inactivating protein that undergoes retrograde transport to the endoplasmic reticulum to exert its cytotoxic effect on eukaryotic cells. Its catalytically active A(1) domain subsequently migrates from the endoplasmic reticulum (ER) lumen to the cytoplasm. To study this final retrotranslocation event, a suicide assay was developed based on the cytoplasmic expression and ER-targeting of the cytotoxic Slt-I A(1) fragment in Saccharomyces cerevisiae. Expression of the Slt-I A(1) domain (residues 1-251) with and without an ER-targeting sequence was lethal to the host and demonstrated that this domain can efficiently migrate from the ER compartment to the cytosol. Deletion analyses revealed that residues 1-239 represent the minimal A(1) segment displaying full enzymatic activity. This fragment, however, accumulates in the ER lumen when directed to this compartment. The addition of residues 240-251 restores the translocation property of the A(1) chain in yeast. However, single mutations within this region do not significantly alter this function in the context of the 251-residue long A(1) domain or affect the toxicity of the resulting Slt-I variants toward Vero cells in the context of the holotoxin. Since this mechanism of retrotranslocation is common to other protein toxins lacking a peptide motif similar in sequence to residues 240-251, the present results suggest that the ER export mechanism may involve the recognition of a more universal structural element, such as a misfolded or altered peptide domain localized at the C terminus of the A(1) chain (residues 240-251) rather than a unique ER export signal sequence. PMID- 15817450 TI - Defects in SUMO (small ubiquitin-related modifier) conjugation and deconjugation alter cell sensitivity to DNA topoisomerase I-induced DNA damage. AB - Eukaryotic DNA topoisomerase I (Top1p) has important functions in DNA replication, transcription, and recombination. This enzyme also constitutes the cellular target of camptothecin (CPT), which induces S-phase-dependent cytotoxicity. To define cellular pathways that regulate cell sensitivity to Top1p induced DNA lesions, we described a yeast genetic screen for conditional tah (top1T722A-hypersensitive) mutants with enhanced sensitivity to low levels of the CPT mimetic mutant top1T722A (Reid, R. J., Fiorani, P., Sugawara, M., and Bjornsti, M. A. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 11440-11445; Fiorani, P., Reid, R. J., Schepis, A., Jacquiau, H. R., Guo, H., Thimmaiah, P., Benedetti, P., and Bjornsti, M. A. (2004) J. Biol. Chem. 279, 21271-21281). Here we report that tah mutant ubc9-10 harbors a hypomorphic allele of UBC9, which encodes the essential SUMO (small ubiquitin-related modifier) E2-conjugating enzyme. The same conditional ubc9P123L mutant was also isolated in an independent screen for enhanced sensitivity to a distinct Top1p poison, Top1N726Hp. The ubc9-10 mutant exhibited a decrease in global protein sumoylation and increased sensitivity to a wide range of DNA-damaging agents independent of Top1p. Deletion of the Ulp2 SUMO protease failed to restore ubc9-10 cell resistance to Top1p poisons or hydroxyurea yet adversely affected wild-type TOP1 cell genetic stability and sensitivity to hydroxyurea. Moreover, although mutation of different consensus SUMO sites in the N terminus and linker region of yeast Top1p failed to recapitulate ubc9-10 mutant phenotypes, they revealed distinct and subtle effects on cell sensitivity to CPT. These results provide insights into the complexities of SUMO conjugation and the confounding effects of SUMO modification on Top1p function and cell sensitivity to genotoxic agents. PMID- 15817451 TI - Caveolin-1 enhances tissue factor pathway inhibitor exposure and function on the cell surface. AB - Tissue factor pathway inhibitor (TFPI) blocks tissue factor-factor VIIa (TF FVIIa) activation of factors X and IX through the formation of the TF-FVIIa-FXa TFPI complex. Most TFPI in vivo associates with caveolae in endothelial cells (EC). The mechanism of this association and the anticoagulant role of caveolar TFPI are not yet known. Here we show that expression of caveolin-1 (Cav-1) in 293 cells keeps TFPI exposed on the plasmalemma surface, decreases the membrane lateral mobility of TFPI, and increases the TFPI-dependent inhibition of TF FVIIa. Caveolae-associated TFPI supports the co-localization of the quaternary complex with caveolae. To investigate the significance of these observations for EC we used RNA interference to deplete the cells of Cav-1. Functional assays and fluorescence microscopy revealed that the inhibitory properties of TFPI were diminished in EC lacking Cav-1, apparently through deficient assembly of the quaternary complex. These findings demonstrate that caveolae regulate the inhibition by cell-bound TFPI of the active protease production by the extrinsic pathway of coagulation. PMID- 15817452 TI - X-ray structure of human acid-beta-glucosidase covalently bound to conduritol-B epoxide. Implications for Gaucher disease. AB - Gaucher disease is an inherited metabolic disorder caused by mutations in the lysosomal enzyme acid-beta-glucosidase (GlcCerase). We recently determined the x ray structure of GlcCerase to 2.0 A resolution (Dvir, H., Harel, M., McCarthy, A. A., Toker, L., Silman, I., Futerman, A. H., and Sussman, J. L. (2003) EMBO Rep.4, 704-709) and have now solved the structure of Glc-Cerase conjugated with an irreversible inhibitor, conduritol-B-epoxide (CBE). The crystal structure reveals that binding of CBE to the active site does not induce a global conformational change in GlcCerase and confirms that Glu340 is the catalytic nucleophile. However, only one of two alternative conformations of a pair of flexible loops (residues 345-349 and 394-399) located at the entrance to the active site in native GlcCerase is observed in the GlcCerase-CBE structure, a conformation in which the active site is accessible to CBE. Analysis of the dynamics of these two alternative conformations suggests that the two loops act as a lid at the entrance to the active site. This possibility is supported by a cluster of mutations in loop 394-399 that cause Gaucher disease by reducing catalytic activity. Moreover, in silico mutational analysis demonstrates that all these mutations stabilize the conformation that limits access to the active site, thus providing a mechanistic explanation of how mutations in this loop result in Gaucher disease. PMID- 15817453 TI - Regulation of macrophage cholesterol efflux through hydroxymethylglutaryl-CoA reductase inhibition: a role for RhoA in ABCA1-mediated cholesterol efflux. AB - The cholesterol biosynthetic pathway produces numerous signaling molecules. Oxysterols through liver X receptor (LXR) activation regulate cholesterol efflux, whereas the non-sterol mevalonate metabolite, geranylgeranyl pyrophosphate (GGPP), was recently demonstrated to inhibit ABCA1 expression directly, through antagonism of LXR and indirectly through enhanced RhoA geranylgeranylation. We used HMG-CoA reductase inhibitors (statins) to test the hypothesis that reduced synthesis of mevalonate metabolites would enhance cholesterol efflux and attenuate foam cell formation. Preincubation of THP-1 macrophages with atorvastatin, dose dependently (1-10 microm) stimulated cholesterol efflux to apolipoprotein AI (apoAI, 10-60%, p < 0.05) and high density lipoprotein (HDL(3)) (2-50%, p < 0.05), despite a significant decrease in cholesterol synthesis (2 90%). Atorvastatin also increased ABCA1 and ABCG1 mRNA abundance (30 and 35%, p < 0.05). Addition of mevalonate, GGPP or farnesyl pyrophosphate completely blocked the statin-induced increase in ABCA1 expression and apoAI-mediated cholesterol efflux. A role for RhoA was established, because two inhibitors of Rho protein activity, a geranylgeranyl transferase inhibitor and C3 exoenzyme, increased cholesterol efflux to apoAI (20-35%, p < 0.05), and macrophage expression of dominant-negative RhoA enhanced cholesterol efflux to apoAI (20%, p < 0.05). In addition, atorvastatin increased the RhoA levels in the cytosol fraction and decreased the membrane localization of RhoA. Atorvastatin treatment activated peroxisome proliferator activated receptor gamma and increased LXR-mediated gene expression suggesting that atorvastatin induces cholesterol efflux through a molecular cascade involving inhibition of RhoA signaling, leading to increased peroxisome proliferator activated receptor gamma activity, enhanced LXR activation, increased ABCA1 expression, and cholesterol efflux. Finally, statin treatment inhibited cholesteryl ester accumulation in macrophages challenged with atherogenic hypertriglyceridemic very low density lipoproteins indicating that statins can regulate foam cell formation. PMID- 15817454 TI - In situ photoactivation of a caged phosphotyrosine peptide derived from focal adhesion kinase temporarily halts lamellar extension of single migrating tumor cells. AB - Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, mediates integrin based cell signaling by transferring signals regulating cell migration, adhesion, and survival from the extracellular matrix to the cytoplasm. Following autophosphorylation at tyrosine 397, FAK binds the Src homology 2 domains of Src and phosphoinositide 3-kinase, among several other possible binding partners. To further investigate the role of phosphorylated FAK in cell migration in situ, peptides comprising residues 391-406 of human FAK with caged phosphotyrosine 397 were synthesized. Although the caged phosphopeptides were stable to phosphatase activity, the free phosphopeptides showed a half-life of approximately 10-15 min in cell lysates. Migrating NBT-II cells (a rat bladder tumor cell line) were microinjected with the caged FAK peptide and locally photoactivated using a focused laser beam. The photoactivation of caged FAK peptide in 8-microm diameter spots over the cell body led to the temporary arrest of the leading edge migration within approximately 1 min of irradiation. In contrast, cell body migration was not inhibited. Microinjection of a non-caged phosphorylated tyrosine 397 FAK peptide into migrating NBT-II cells also led to lamellar arrest; however, this approach lacks the temporal control afforded by the caged phosphopeptides. Photoactivation of related phosphotyrosine peptides with altered sequences did not result in transient lamellar arrest. We hypothesize that the phosphorylated FAK peptide competes with the endogenous FAK for binding to FAK effectors including, but not limited to, Src and phosphoinositide 3-kinase, causing spatiotemporal misregulation and subsequent lamellar arrest. PMID- 15817455 TI - The N-acetyl-D-glucosaminylphosphatidylinositol De-N-acetylase of glycosylphosphatidylinositol biosynthesis is a zinc metalloenzyme. AB - The de-N-acetylation of N-acetyl-D-glucosaminylphosphatidylinositol (GlcNAc-PI) is the second step of mammalian and trypanosomal glycosylphosphatidylinositol biosynthesis. Glycosylphosphatidylinositol biosynthesis is essential for Trypanosoma brucei, the causative agent of African sleeping sickness, and GlcNAc PI de-N-acetylase has previously been validated as a drug target. Inhibition of the trypanosome cell-free system and recombinant rat GlcNAc-PI de-N-acetylase by divalent metal cation chelators demonstrates that a tightly bound divalent metal cation is essential for activity. Reconstitution of metal-free GlcNAc-PI de-N acetylase with divalent metal cations restores activity in the order Zn(2+) > Cu(2+) > Ni(2+) > Co(2+) > Mg(2+). Site-directed mutagenesis and homology modeling were used to identify active site residues and postulate a mechanism of action. The characterization of GlcNAc-PI de-N-acetylase as a zinc metalloenzyme will facilitate the rational design of anti-protozoan parasite drugs. PMID- 15817456 TI - A novel dimeric structure of the RimL Nalpha-acetyltransferase from Salmonella typhimurium. AB - RimL is responsible for converting the prokaryotic ribosomal protein from L12 to L7 by acetylation of its N-terminal amino group. We demonstrate that purified RimL is capable of posttranslationally acetylating L12, exhibiting a V(max) of 21 min(-1). We have also determined the apostructure of RimL from Salmonella typhimurium and its complex with coenzyme A, revealing a homodimeric oligomer with structural similarity to other Gcn5-related N-acetyltransferase superfamily members. A large central trough located at the dimer interface provides sufficient room to bind both L12 N-terminal helices. Structural and biochemical analysis indicates that RimL proceeds by single-step transfer rather than a covalent-enzyme intermediate. This is the first structure of a Gcn5-related N acetyltransferase family member with demonstrated activity toward a protein N(alpha)-amino group and is a first step toward understanding the molecular basis for N(alpha)acetylation and its function in cellular regulation. PMID- 15817457 TI - DNA polymerase kappa is specifically required for recovery from the benzo[a]pyrene-dihydrodiol epoxide (BPDE)-induced S-phase checkpoint. AB - Previously we identified an intra-S-phase cell cycle checkpoint elicited by the DNA-damaging carcinogen benzo[a]pyrene-dihydrodiol epoxide (BPDE). Here we have investigated the roles of lesion bypass DNA polymerases polkappa and poleta in the BPDE-induced S-phase checkpoint. BPDE treatment induced the re-localization of an ectopically expressed green fluorescent protein-polkappa fusion protein to nuclear foci containing sites of active DNA synthesis in human lung carcinoma H1299 cells. In contrast, a similarly expressed yellow fluorescent protein-poleta fusion protein showed a constitutive nuclear focal distribution at replication forks (in the same cells) that was unchanged in response to BPDE. BPDE-induced formation of green fluorescent protein-polkappa nuclear foci was temporally coincident with checkpoint-mediated S-phase arrest. Unlike "wild-type" cells, Polk(-/-) mouse embryonic fibroblasts (MEFs) failed to recover from BPDE-induced S-phase arrest, while exhibiting normal recovery from S-phase arrest induced by ionizing radiation and hydroxyurea. XPV fibroblasts lacking poleta showed a normal S-phase checkpoint response to BPDE (but failed to recover from the UV light-induced S-phase checkpoint), in sharp contrast to Polk(-/-) MEFs. The persistent S-phase arrest in BPDE-treated Polk(-/-) cells was associated with increased levels of histone gammaH2AX (a marker of DNA double-strand breaks (DSBs)) and activation of the DSB-responsive kinases ATM and Chk2. These data suggest that in the absence of polkappa, replication forks stall at sites of damage and collapse and generate DSBs. Therefore, we conclude that the trans lesion synthesis enzyme polkappa is specifically required for normal recovery from the BPDE-induced S-phase checkpoint. PMID- 15817458 TI - 3-hydroxykynurenine-mediated modification of human lens proteins: structure determination of a major modification using a monoclonal antibody. AB - Tryptophan can be oxidized in the eye lens by both enzymatic and non-enzymatic mechanisms. Oxidation products, such as kynurenines, react with proteins to form yellow-brown pigments and cause covalent cross-linking. We generated a monoclonal antibody against 3-hydroxykynurenine (3OHKYN)-modified keyhole limpet hemocyanin and characterized it using 3OHKYN-modified amino acids and proteins. This monoclonal antibody reacted with 3OHKYN-modified N(alpha)-acetyl lysine, N(alpha) acetyl histidine, N(alpha)-acetyl arginine, and N(alpha)-acetyl cysteine. Among the several tryptophan oxidation products tested, 3OHKYN produced the highest concentration of antigen when reacted with human lens proteins. A major antigen from the reaction of 3OHKYN and N(alpha)-acetyl lysine was purified by reversed phase high pressure liquid chromatography, which was characterized by spectroscopy and identified as 2-amino-3-hydroxyl-alpha-((5S)-5-acetamino-5 carboxypentyl amino)-gamma-oxo-benzene butanoic acid. Enzyme-digested cataractous lens proteins displayed 3OHKYN-derived modifications. Immunohistochemistry revealed 3OHKYN modifications in proteins associated with the lens fiber cell plasma membrane. The low molecular products (<10,000 Da) isolated from normal lenses after reaction with glucosidase followed by incubation with proteins generated 3OHKYN-derived products. Human lens epithelial cells incubated with 3OHKYN showed intense immunoreactivity. We also investigated the effect of glycation on tryptophan oxidation and kynurenine-mediated modification of lens proteins. The results showed that glycation products failed to oxidize tryptophan or generate kynurenine modifications in proteins. Our studies indicate that 3OHKYN modifies lens proteins independent of glycation to form products that may contribute to protein aggregation and browning during cataract formation. PMID- 15817459 TI - Down-regulation of human DAB2IP gene expression mediated by polycomb Ezh2 complex and histone deacetylase in prostate cancer. AB - Human DAB2IP (hDAB2IP), a novel GTPase-activating protein modulating the Ras mediated signaling and tumor necrosis factor-mediated apoptosis, is a potent growth inhibitor in human prostate cancer (PCa). Loss of hDAB2IP expression in PCa is due to altered epigenetic regulation (i.e. DNA methylation and histone modification) of its promoter region. The elevated polycomb Ezh2, a histone methyltransferase, has been associated with PCa progression. In this study, we have demonstrated that an increased Ezh2 expression in normal prostatic epithelial cells can suppress hDAB2IP gene expression. In contrast, knocking down the endogenous Ezh2 levels in PCa by a specific small interfering RNA can increase hDAB2IP expression. The association of Ezh2 complex (including Eed and Suz12) with hDAB2IP gene promoter is also detected in PCa cells but not in normal prostatic epithelial cells. Increased Ezh2 expression in normal prostatic epithelial cells by cDNA transfection facilitates the recruitment of other components of Ezh2 complex to the hDAB2IP promoter region accompanied with the increased levels of methyl histone H3 (H3) and histone deacetylase (HDAC1). Consistently, data from PCa cells transfected with Ezh2 small interfering RNA demonstrated that reduced Ezh2 levels resulted in the dissociation of Ezh2 complex accompanied with decreased levels of both methyl H3 and HDAC1 from hDAB2IP gene promoter. We further unveiled that the methylation status of Lys-27 but not Lys-9 of H3 in hDAB2IP promoter region is consistent with the hDAB2IP levels in both normal prostatic epithelial cells and PCa cells. Together, we conclude that hDAB2IP gene is a target gene of Ezh2 in prostatic epithelium, which provides an underlying mechanism of the down-regulation of hDAB2IP gene in PCa. PMID- 15817460 TI - Endo180 binds to the C-terminal region of type I collagen. AB - Type I collagen is a fibril-forming heterotrimer composed of two alpha1 and one alpha2 chains and plays a crucial role in cell-matrix adhesion and cell differentiation. Through a comprehensive differential display screening of oncogenic ras target genes, we have shown that the alpha1 chain of type I collagen (col1a1) is markedly down-regulated by the ras oncogene through the mitogen-activated protein kinase pathway. Although ras-transformed cells are no longer able to produce and secrete endogenous collagen, they can still adhere to exogenous collagen, suggesting that the cells express a collagen binding factor(s) on the cell surface. When the region of col1a1 encompassing the C terminal glycine repeat and C-prodomain (amino acids 1000-1453) was affinity labeled with human placental alkaline phosphatase, the secreted trimeric fusion protein could bind to the surface of Ras-transformed cells. Using biochemical purification followed by matrix-assisted laser desorption/ionization mass spectrometry analysis, we identified this collagen binding factor as Endo180 (uPARAP, CD280), a member of the mannose receptor family. Ectopic expression of Endo180 in CosE5 cells followed by in situ staining and quantitative binding assays confirmed that Endo180 indeed recognizes and binds to placental alkaline phosphatase. The interaction between Endo180 and the C-terminal region of type I collagen appears to play an important role in cell-matrix adhesion. PMID- 15817461 TI - Functional properties of the p33 and p55 domains of the Helicobacter pylori vacuolating cytotoxin. AB - Helicobacter pylori secretes an 88-kDa vacuolating cytotoxin (VacA) that may contribute to the pathogenesis of peptic ulcer disease and gastric cancer. VacA cytotoxic activity requires assembly of VacA monomers into oligomeric structures, formation of anion-selective membrane channels, and entry of VacA into host cells. In this study, we analyzed the functional properties of recombinant VacA fragments corresponding to two putative VacA domains (designated p33 and p55). Immunoprecipitation experiments indicated that these two domains can interact with each other to form protein complexes. In comparison to the individual VacA domains, a mixture of the p33 and p55 proteins exhibited markedly enhanced binding to the plasma membrane of mammalian cells. Furthermore, internalization of the VacA domains was detected when cells were incubated with the p33/p55 mixture but not when the p33 and p55 proteins were tested individually. Incubation of cells with the p33/p55 mixture resulted in cell vacuolation, whereas the individual domains lacked detectable cytotoxic activity. Interestingly, sequential addition of p55 followed by p33 resulted in VacA internalization and cell vacuolation, whereas sequential addition in the reverse order was ineffective. These results indicate that both the p33 and p55 domains contribute to the binding and internalization of VacA and that both domains are required for vacuolating cytotoxic activity. Reconstitution of toxin activity from two separate domains, as described here for VacA, has rarely been described for pore-forming bacterial toxins, which suggests that VacA is a pore-forming toxin with unique structural properties. PMID- 15817462 TI - The forkhead box m1 transcription factor is essential for embryonic development of pulmonary vasculature. AB - Transgenic and gene knock-out studies demonstrated that the mouse Forkhead Box m1 (Foxm1 or Foxm1b) transcription factor (previously called HFH-11B, Trident, Win, or MPP2) is essential for hepatocyte entry into mitosis during liver development, regeneration, and liver cancer. Targeted deletion of Foxm1 gene in mice produces an embryonic lethal phenotype due to severe abnormalities in the development of liver and heart. In this study, we show for the first time that Foxm1(-/-) lungs exhibit severe hypertrophy of arteriolar smooth muscle cells and defects in the formation of peripheral pulmonary capillaries as evidenced by significant reduction in platelet endothelial cell adhesion molecule 1 staining of the distal lung. Consistent with these findings, significant reduction in proliferation of the embryonic Foxm1(-/-) lung mesenchyme was found, yet proliferation levels were normal in the Foxm1-deficient epithelial cells. Severe abnormalities of the lung vasculature in Foxm1(-/-) embryos were associated with diminished expression of the transforming growth factor beta receptor II, a disintegrin and metalloprotease domain 17 (ADAM-17), vascular endothelial growth factor receptors, Polo-like kinase 1, Aurora B kinase, laminin alpha4 (Lama4), and the Forkhead Box f1 transcription factor. Cotransfection studies demonstrated that Foxm1 stimulates transcription of the Lama4 promoter, and this stimulation requires the Foxm1 binding sites located between -1174 and -1145 bp of the mouse Lama4 promoter. In summary, development of mouse lungs depends on the Foxm1 transcription factor, which regulates expression of genes essential for mesenchyme proliferation, extracellular matrix remodeling, and vasculogenesis. PMID- 15817463 TI - Actopaxin interacts with TESK1 to regulate cell spreading on fibronectin. AB - The focal adhesion protein actopaxin contributes to integrin-actin associations and is involved in cell adhesion, spreading, and motility. Herein, we identify and characterize an association between actopaxin and the serine/threonine kinase testicular protein kinase 1 (TESK1), a ubiquitously expressed protein previously reported to regulate cellular spreading and focal adhesion formation via phosphorylation of cofilin. The interaction between actopaxin and TESK1 is direct and the binding sites were mapped to the carboxyl terminus of both proteins. The association between actopaxin and TESK1 is negatively regulated by adhesion to fibronectin, and a phosphomimetic actopaxin mutant that promotes cell spreading also exhibits impaired binding to TESK1. Binding of actopaxin to TESK1 inhibits TESK1 kinase activity in vitro. Expression of the carboxyl terminus of actopaxin has previously been reported to retard cell spreading. This effect was reversed following overexpression of TESK1 and was found to be dependent on an inability of actopaxin carboxyl terminus expressing cells to promote cofilin phosphorylation upon matrix adhesion and caused by retention of TESK1 by this actopaxin mutant. Thus, the association between actopaxin and TESK1, which is likely regulated by phosphorylation of actopaxin, regulates TESK1 activity and subsequent cellular spreading on fibronectin. PMID- 15817464 TI - Glucose-dependent insulinotropic polypeptide (GIP) stimulation of pancreatic beta cell survival is dependent upon phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signaling, inactivation of the forkhead transcription factor Foxo1, and down-regulation of bax expression. AB - The hormone glucose-dependent insulinotropic polypeptide (GIP) potently stimulates insulin secretion and promotes beta-cell proliferation and cell survival. In the present study we identified Forkhead (Foxo1)-mediated suppression of the bax gene as a critical component of the effects of GIP on cell survival. Treatment of INS-1(832/13) beta-cells with GIP resulted in concentration-dependent activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB)/Foxo1 signaling module. In parallel studies, GIP decreased bax promoter activity. Serial deletion analysis of the bax promoter demonstrated that the region -682 to -320, containing FHRE-II (5AAAACAAACA), was responsible for GIP-mediated effects. Foxo1 bound to FHRE-II in gel mobility shift assays, and Foxo1-FHRE-II interactions conferred GIP responsiveness to the bax promoter. INS-1 cells incubated under proapoptotic and glucolipotoxic conditions demonstrated increased nuclear localization of Foxo1 and bax promoter activity and decreased cytoplasmic phospho-PKB/Foxo1. GIP partially restored expression PKB/Foxo1 and bax promoter activity. Similar protective effects were found with dispersed islet cells from C57BL/6 mice, but not with those from GIP receptor knock-out (GIPR(-/-)) mice. GIP treatment reduced glucolipotoxicity induced cell death in C57 BL/6 and Bax(-/-) islets, but not GIPR(-/-) mouse islets. Chronic treatment of Vancouver diabetic fatty Zucker rats with GIP resulted in down-regulation of Bax and up-regulation of Bcl-2 in pancreatic beta cells. The results show that PI3K/PKB/Foxo1 signaling mediates GIP suppression of bax gene expression and that this module is a key pathway by which GIP regulates beta-cell apoptosis in vivo. PMID- 15817465 TI - Insights into hydrophobicity and the chaperone-like function of alphaA- and alphaB-crystallins: an isothermal titration calorimetric study. AB - Alpha-crystallin, composed of two subunits, alphaA and alphaB, has been shown to function as a molecular chaperone that prevents aggregation of other proteins under stress conditions. The exposed hydrophobic surfaces of alpha-crystallins have been implicated in this process, but their exact role has not been elucidated. In this study, we quantify the hydrophobic surfaces of alphaA- and alphaB-crystallins by isothermal titration calorimetry using 8-anilino-1 napthalenesulfonic acid (ANS) as a hydrophobic probe and analyze its correlation to the chaperone potential of alphaA- and alphaB-crystallins under various conditions. Two ANS binding sites, one with low and another with high affinity, were clearly detected, with alphaB showing a higher number of sites than alphaA at 30 degrees C. In agreement with the higher number of hydrophobic sites, alphaB crystallin demonstrated higher chaperone activity than alphaA at this temperature. Thermodynamic analysis of ANS binding to alphaA- and alphaB crystallins indicates that high affinity binding is driven by both enthalpy and entropy changes, with entropy dominating the low affinity binding. Interestingly, although the number of ANS binding sites was similar for alphaA and alphaB at 15 degrees C, alphaA was more potent than alphaB in preventing aggregation of the insulin B-chain. Although there was no change in the number of high affinity binding sites of alphaA and alphaB for ANS upon preheating, there was an increase in the number of low affinity sites of alphaA and alphaB. Preheated alphaA, in contrast to alphaB, exhibited remarkably enhanced chaperone activity. Our results indicate that although hydrophobicity appears to be a factor in determining the chaperone-like activity of alpha-crystallins, it does not quantitatively correlate with the chaperone function of alpha-crystallins. PMID- 15817466 TI - Identification of amino acids required for the functional up-regulation of human dihydrofolate reductase protein in response to antifolate Treatment. AB - Human dihydrofolate reductase (DHFR) protein levels rapidly increase upon exposure to methotrexate, a potent inhibitor of this enzyme. A model to explain this increase proposes that DHFR inhibits its own translation by binding to its cognate mRNA and that methotrexate disrupts the DHFR protein-mRNA complex allowing its translation to resume. In the present study, Chinese hamster ovary cells lacking DHFR were transfected with wild type and mutants of human DHFR to identify amino acids that are essential for increases in DHFR in response to methotrexate. Glu-30, Leu-22, and Ser-118 were involved in the up-regulation of DHFR protein levels by methotrexate and certain other antifolates. Cells transfected with E30A, L22R, and S118A mutants that did not respond to methotrexate up-regulation had higher basal levels of DHFR, consistent with the model, i.e. lack of feedback regulation of these enzymes. Although cells containing the S118A mutant enzyme had higher levels of DHFR and had catalytic activity similar to that of wild type DHFR, they had the same sensitivity to the cytotoxicity of methotrexate, as were cells with wild type DHFR. This finding provides evidence that the adaptive up-regulation of DHFR by methotrexate contributes to the decreased sensitivity to this drug. Based on these observations, a new model is proposed whereby DHFR exists in two conformations, one bound to DHFR mRNA and the other bound to NADPH. The mutants that are not up regulated by methotrexate are unable to bind their cognate mRNA. PMID- 15817467 TI - Graded levels of GATA-1 expression modulate survival, proliferation, and differentiation of erythroid progenitors. AB - Transcription factor GATA-1 plays an important role in gene regulation during the development of erythroid cells. Several reports suggest that GATA-1 plays multiple roles in survival, proliferation, and differentiation of erythroid cells. However, little is known about the relationship between the level of GATA 1 expression and its nature of multifunction to affect erythroid cell fate. To address this issue, we developed in vitro embryonic stem (ES) culture system by using OP9 stromal cells (OP9/ES cell co-culture system), and cultured the mutant (GATA-1.05 and GATA-1-null) and wild type (WT)ES cells, respectively. By using this OP9/ES cell co-culture system, primitive and definitive erythroid cells were developed individually, and we examined how expression level of GATA-1 affects the development of erythroid cells. GATA-1.05 ES-derived definitive erythroid cells were immature with the appearance of proerythroblasts, and highly proliferated, compared with WT and GATA-1-null ES-derived erythroid cells. Extensive studies of cell cycle kinetics revealed that the GATA-1.05 proerythroblasts accumulated in S phase and expressed lower levels of p16(INK4A) than WT ES cell-derived proerythroblasts. We concluded that GATA-1 must achieve a critical threshold activity to achieve selective activation of specific target genes, thereby influencing the developmental decision of an erythroid progenitor cell to undergo apoptosis, proliferation, or terminal differentiation. PMID- 15817468 TI - The glucagon-like peptide-2 receptor C terminus modulates beta-arrestin-2 association but is dispensable for ligand-induced desensitization, endocytosis, and G-protein-dependent effector activation. AB - Classic models of receptor desensitization and internalization have been largely based on the behavior of Family A G-protein-coupled receptors (GPCRs). The glucagon-like peptide-2 receptor (GLP-2R) is a member of the Family B glucagon secretin GPCR family, which exhibit significant sequence and structural differences from the Family A receptors in their intracellular and extracellular domains. To identify structural motifs that regulate GLP-2R signaling and cell surface receptor expression, we analyzed the functional properties of a series of mutant GLP-2Rs. The majority of the C-terminal receptor tail was dispensable for GLP-2-induced cAMP accumulation, ERK1/2 activation, and endocytosis in transfected cells. However, progressive truncation of the C terminus reduced cell surface receptor expression, altered agonist-induced GLP-2R trafficking, and abrogated protein kinase A-mediated heterologous receptor desensitization. Elimination of the distal 21 amino acids of the receptor was sufficient to promote constitutive receptor internalization and prevent agonist-induced recruitment of beta-arrestin-2. Site-directed mutagenesis identified specific amino acid residues within the distal GLP-2R C terminus that mediate the stable association with beta-arrestin-2. Surprisingly, although the truncated mutant receptors failed to interact with beta-arrestin-2, they underwent homologous desensitization and subsequent resensitization with kinetics similar to that observed with the wild-type GLP-2R. Our data suggest that, although the GLP-2R C terminus is not required for coupling to cellular machinery regulating signaling or desensitization, it may serve as a sorting signal for intracellular trafficking. Taken together with the previously demonstrated clathrin and dynamin independent, lipid-raft-dependent pathways for internalization, our data suggest that GLP-2 receptor signaling has evolved unique structural and functional mechanisms for control of receptor trafficking, desensitization, and resensitization. PMID- 15817469 TI - Regulation of inflammatory response in neural cells in vitro by thiadiazolidinones derivatives through peroxisome proliferator-activated receptor gamma activation. AB - In most neurodegenerative disorders, including multiple sclerosis, Parkinson disease, and Alzheimer disease, a massive neuronal cell death occurs as a consequence of an uncontrolled inflammatory response, where activated astrocytes and microglia and their cytotoxic agents play a crucial pathological role. Current treatments for these diseases are not effective. In the present study we investigate the effect of thiadiazolidinone derivatives, which have been recently suggested to play a role in neurodegenerative disorders. We have found that thiadiazolidinones are potent neuroprotector compounds. Thiadiazolidinones inhibited inflammatory activation of cultured brain astrocytes and microglia by diminishing lipopolysaccharide-induced interleukin 6, tumor necrosis factor alpha, inducible nitric-oxide synthase, and inducible cyclooxygenase type 2 expression. In addition, thiadiazolidinones inhibited tumor necrosis factor-alpha and nitric oxide production and, concomitantly, protected cortical neurons from cell death induced by the cell-free supernatant from activated microglia. The neuroprotective effects of thiadiazolidinones are completely inhibited by the peroxisome proliferator-activated receptor gamma antagonist GW9662. In contrast the glycogen synthase kinase 3beta inhibitor LiCl did not show any effect. These findings suggest that thiadiazolidinones potently attenuate lipopolysaccharide induced neuroinflammation and reduces neuronal death by a mechanism dependent of peroxisome proliferator-activated receptor gamma activation. PMID- 15817470 TI - Functional analyses of oxygenases in jadomycin biosynthesis and identification of JadH as a bifunctional oxygenase/dehydrase. AB - A novel angucycline metabolite, 2,3-dehydro-UWM6, was identified in a jadH mutant of Streptomyces venezuelae ISP5230. Both UWM6 and 2,3-dehydro-UWM6 could be converted to jadomycin A or B by a ketosynthase alpha (jadA) mutant of S. venezuelae. These angucycline intermediates were also converted to jadomycin A by transformant of the heterologous host Streptomyces lividans expressing the jadFGH oxygenases in vivo and by its cell-free extracts in vitro; thus the three gene products JadFGH are implicated in catalysis of the post-polyketide synthase biosynthetic reactions converting UWM6 to jadomycin aglycone. Genetic and biochemical analyses indicate that JadH possesses dehydrase activity, not previously associated with polyketide-modifying oxygenase. Since the formation of aromatic polyketides often requires multiple dehydration steps, bifunctionality of oxygenases modifying aromatic polyketides may be a general phenomenon. PMID- 15817471 TI - Degradation of the tumor suppressor Smad4 by WW and HECT domain ubiquitin ligases. AB - Smad4 mediates signaling by the transforming growth factor-beta (TGF-beta) superfamily of cytokines. Smad signaling is negatively regulated by inhibitory (I) Smads and ubiquitin-mediated processes. Known mechanisms of proteasomal degradation of Smads depend on the direct interaction of specific E3 ligases with Smads. Alternatively, I-Smads elicit degradation of the TGF-beta receptor by recruiting the WW and HECT domain E3 ligases, Smurfs, WWP1, or NEDD4-2. We describe an equivalent mechanism of degradation of Smad4 by the above E3 ligases, via formation of ternary complexes between Smad4 and Smurfs, mediated by R-Smads (Smad2) or I-Smads (Smad6/7), acting as adaptors. Smurfs, which otherwise cannot directly bind to Smad4, mediated poly-ubiquitination of Smad4 in the presence of Smad6 or Smad7. Smad4 co-localized with Smad7 and Smurf1 primarily in the cytoplasm and in peripheral cell protrusions. Smad2 or Smad7 mutants defective in Smad4 interaction failed to induce Smurf1-mediated down-regulation of Smad4. A Smad4 mutant defective in Smad2 or Smad7 interaction could not be effectively down-regulated by Smurf1. We propose that Smad4 is targeted for degradation by multiple ubiquitin ligases that can simultaneously act on R-Smads and signaling receptors. Such mechanisms of down-regulation of TGF-beta signaling may be critical for proper physiological response to this pathway. PMID- 15817472 TI - Differentiation of epithelial Na+ channel function. An in vitro model. AB - Confluent monolayers of epithelial cells grown on nonporous support form fluid filled hemicysts called domes, which reflect active ion transport across the epithelium. Clara-like H441 lung adenocarcinoma cells grown on glass supports and exposed to 50 nM dexamethasone developed domes in a time-dependent fashion. Uplifting of small groups of cells occurred within 6-12 h, well formed domes appeared between 24 and 48 h, and after 7 days, individual domes started to merge. Cells inside of domes compared with those outside domes, or with monolayers not exposed to dexamethasone, differed by higher surfactant production, an increased cytokeratin expression, and the localization of claudin 4 proteins to the plasma membrane. In patch clamp studies, amiloride-blockable sodium currents were detected exclusively in cells inside domes, whereas in cells outside of domes, sodium crossed the membrane through La3+-sensitive nonspecific cation channels. Cells grown on permeable support without dexamethasone expressed amiloride-sensitive currents only after tight electrical coupling was achieved (transepithelial electrical resistance (R(t)) > 1 kilohm). In real-time quantitative PCR experiments, the addition of dexamethasone increased the content of claudin-4, occludin, and Na+ channel gamma-subunit (gamma-ENaC) mRNAs by 1.34 , 1.32-, and 1.80-fold, respectively, after 1 h and was followed by an increase at 6 h in the content of mRNA of alpha- and beta-ENaC and of alpha1- and beta1 Na,K-ATPase. In the absence of dexamethasone, neither change in gene expression nor cell uplifting was observed. Our data suggest that during epithelial differentiation, coordinated expression of tight junction proteins precedes the development of vectorial transport of sodium, which in turn leads to the fluid accumulation in basolateral spaces that is responsible for dome formation. PMID- 15817473 TI - Nuclear localization of flavonoid enzymes in Arabidopsis. AB - Flavonoids represent one of the oldest, largest, and most diverse families of plant secondary metabolites. These compounds serve a wide range of functions in plants, from pigmentation and UV protection to the regulation of hormone transport. Flavonoids also have interesting pharmacological activities in animals that are increasingly being characterized in terms of effects on specific proteins or other macromolecules. Although flavonoids are found in many different locations both inside and outside the cell, biosynthesis has long been believed to take place exclusively in the cytoplasm. Recent reports from a number of different plant species have documented the presence of flavonoids in nuclei, raising the possibility of novel mechanisms of action for these compounds. Here we present evidence that not only flavonoids, but also at least two of the biosynthetic enzymes, are located in the nucleus in several cell types in Arabidopsis. This is the first indication that differential targeting of the biosynthetic machinery may be used to regulate the deposition of plant secondary products at diverse sites of action within the cell. PMID- 15817474 TI - Synthesis of sphingolipids with very long chain fatty acids but not ergosterol is required for routing of newly synthesized plasma membrane ATPase to the cell surface of yeast. AB - The proton pumping H(+)-ATPase, Pma1p, is an abundant and very long-lived polytopic protein of the Saccharomyces cerevisiae plasma membrane. Pma1p constitutes a major cargo of the secretory pathway and thus serves as an excellent model to study plasma membrane biogenesis. We have previously shown that newly synthesized Pma1p is mistargeted to the vacuole in an elo3Delta mutant that affects the synthesis of the ceramide-bound C26 very long chain fatty acid (Eisenkolb, M., Zenzmaier, C., Leitner, E., and Schneiter, R. (2002) Mol. Biol. Cell 13, 4414-4428) and now describe a more detailed analysis of the role of lipids in Pma1p biogenesis. Remarkably, a block at various steps of sterol biosynthesis, a complete block in sterol synthesis, or the substitution of internally synthesized ergosterol by externally supplied ergosterol or even by cholesterol does not affect Pma1p biogenesis or its association with detergent resistant membrane domains (lipid "rafts"). However, a block in sphingolipid synthesis or any perturbation in the synthesis of the ceramide-bound C26 very long chain fatty acid results in mistargeting of newly synthesized Pma1p to the vacuole. Mistargeting correlates with a lack of newly synthesized Pma1p to acquire detergent resistance, suggesting that sphingolipids with very long acyl chains affect sorting of Pma1p to the cell surface. PMID- 15817475 TI - Surface calreticulin mediates muramyl dipeptide-induced apoptosis in RK13 cells. AB - Calreticulin (CRT) is a binding protein for apoptotic N-acetylmuramyl-L-alanyl-D isoglutamine (L,D-MDP) or peptidoglycan in RK(13) cells. CRT on RK(13) cell surface (srCRT) forms complex(es) with tumor necrosis factor receptor 1 (TNFR1) and TNFR-associated death domain (TRADD) protein of the cell membrane. CRT polyclonal or monoclonal antibody binding to RK(13) srCRT dose-dependently inhibited L,D-MDP-induced apoptosis. In RK(13) cells, L,D-MDP up-regulated the TNFR1.TRADD complex of the plasma membrane and subsequently induced cytosolic TRADD-Fas-associated death domain protein complex. Biotinylated srCRT was capable of calcium-dependent binding of Sepharose-immobilized L,D-MDP or peptidoglycan. However, Toll-like receptors TLR-2 and TLR-4, Nod2, and CD14 of RK(13) cells did not specifically bind Sepharose-immobilized L,D-MDP. High concentrations (5-40 mm) of EGTA dose-dependently inhibited free L,D-MDP binding to purified RK(13) cell CRT and promoted free L,D-MDP dissociation from RK(13) cell CRT.MDP complex. Different concentrations of EGTA (0-40 mm) added to Dulbecco's modified essential medium with 1.8 mm calcium or phosphate-buffered saline with 0.18 mm calcium have different effects on medium free calcium concentrations but have identical inhibiting effects on L,D-MDP-induced apoptosis. More inhibition of the L,D-MDP induced apoptotic DNA ladders and caspase-3 activity in RK(13) cells was obtained with EGTA pretreatment (83%) than just EGTA + L,D-MDP (47%). The knocking down of srCRT by antisense oligonucleotide CRTAS121 (250 nmol/ml) and stealth small interfering RNA CRT_siR479 (150 pm/ml) for 2 days (44 and 66%, respectively), resulted in the inhibition of L,D-MDP-induced caspase-3 activity (47 and 65%, respectively). The results suggest that (a) the binding of L,D-MDP to srCRT is calcium-dependent, i.e. on srCRT-bound calcium, and (b) it is srCRT, not TLR-2, TLR-4, Nod2 or CD14, that mediates L,D-MDP-induced RK(13) cell apoptosis through activating the TNFR1. TRADD-Fas-associated death domain protein apoptotic pathway. PMID- 15817476 TI - p130cas but not paxillin is essential for Caco-2 intestinal epithelial cell spreading and migration on collagen IV. AB - We have previously observed that collagen IV regulates Caco-2 intestinal epithelial cell spreading and migration via Src kinase and stimulates Src dependent tyrosine phosphorylation of p130cas. We observed that collagen IV also stimulated Src-dependent phosphorylation of both paxillin Tyr31 and paxillin Tyr118. Caco-2 transfection with paxillin or p130cas siRNAs inhibited expression of these proteins by more than 90% for at least 5 days after transfection. Although p130cas siRNA inhibited cell spreading on collagen IV by 33%, three different paxillin siRNAs did not inhibit cell spreading. p130cas siRNA did not affect Src Tyr416 or Src Tyr527 phosphorylation, FAK Tyr397 phosphorylation, or Src-dependent phosphorylation of FAK Tyr925, suggesting that p130cas did not inhibit cell spreading by altering FAK or Src activity. Rat p130cas expression after siRNA knock-out of endogenous human p130cas in Caco-2 cells reduced cell spreading inhibition by 71%. In contrast, expression of rat p130cas from which the Src-phosphorylated substrate domain was deleted did not rescue siRNA inhibition of cell spreading. Combined treatment with siRNAs to Crk and CrkL, which bind to the p130cas substrate domain, inhibited cell spreading by 54%. Both p130cas siRNA and the combined Crk/CrkL siRNAs strongly inhibited (52 and 46% inhibition, respectively) Caco-2 sheet migration on collagen IV and noticeably inhibited lamellipodial extension, whereas paxillin siRNA only inhibited migration by 18% and did not noticeably affect lamellipodial extension. These results suggest that Src may regulate Caco-2 migration on collagen IV via both p130cas and paxillin but that Src phosphorylation of p130cas is more important for this process. PMID- 15817477 TI - Dynamic regulation of Tec kinase localization in membrane-proximal vesicles of a T cell clone revealed by total internal reflection fluorescence and confocal microscopy. AB - Tec family tyrosine kinases are key regulators of lymphocyte activation and effector function. Several Tec family kinases (Tec, Itk, Rlk/Txk) are expressed in T cells, but it is still not clear to what degree these are redundant or have unique functions. We recently demonstrated that Tec alone, among the Tec kinase family members examined, can induce nuclear factor of activated T cell-dependent transcription. This unique functional characteristic correlated with a unique pattern of subcellular localization, as Tec (but not other family members) was found in small vesicles, the appearance of which requires signaling through the T cell receptor for antigen. Here we report on our studies of these Tec-containing structures in live T cells, using total internal reflection fluorescence microscopy. With this technique, we showed that, in live T cells, the Tec vesicles are located at the plasma membrane, the vesicles are unique to Tec (and not the related kinase Itk), and their formation and maintenance require T cell receptor signaling through Src family kinases and PI 3-kinase. Finally, we have imaged isolated T cell membranes by confocal microscopy, confirming the membrane proximal location of Tec vesicles, as well as demonstrating overlap of these vesicles with the tyrosine kinase Lck, the Tec substrate PLC-gamma1, and the early endosomal antigen 1 marker EEA1. PMID- 15817478 TI - Reversible inhibition of alpha-synuclein fibrillization by dopaminochrome mediated conformational alterations. AB - Previous studies demonstrated that alpha-synuclein (alpha-syn) fibrillization is inhibited by dopamine, and studies to understand the molecular basis of this process were conducted (Conway, K. A., Rochet, J. C., Bieganski, R. M., and Lansbury, P. T., Jr. (2001) Science 294, 1346-1349). Dopamine inhibition of alpha syn fibrillization generated exclusively spherical oligomers that depended on dopamine autoxidation but not alpha-syn oxidation, because mutagenesis of Met, His, and Tyr residues in alpha-syn did not abrogate this inhibition. However, truncation of alpha-syn at residue 125 restored the ability of alpha-syn to fibrillize in the presence of dopamine. Mutagenesis and competition studies with specific synthetic peptides identified alpha-syn residues 125-129 (i.e. YEMPS) as an important region in the dopamine-induced inhibition of alpha-syn fibrillization. Significantly, the dopamine oxidation product dopaminochrome was identified as a specific inhibitor of alpha-syn fibrillization. Dopaminochrome promotes the formation of spherical oligomers by inducing conformational changes, as these oligomers regained the ability to fibrillize by simple denaturation/renaturation. Taken together, these data indicate that dopamine inhibits alpha-syn fibrillization by inducing structural changes in alpha-syn that can occur through the interaction of dopaminochrome with the 125YEMPS129 motif of alpha-syn. These results suggest that the dopamine autoxidation can prevent alpha-syn fibrillization in dopaminergic neurons through a novel mechanism. Thus, decreased dopamine levels in substantia nigra neurons might promote alpha-syn aggregation in Parkinson's disease. PMID- 15817479 TI - Bcl-2 rescues ceramide- and etoposide-induced mitochondrial apoptosis through blockage of caspase-2 activation. AB - Recent studies indicate that caspase-2 is involved in the early stage of apoptosis before mitochondrial damage. Although the activation of caspase-2 has been shown to occur in a large protein complex, the mechanisms of caspase-2 activation remain unclear. Here we report a regulatory role of Bcl-2 on caspase-2 upstream of mitochondria. Stress stimuli, including ceramide and etoposide, caused caspase-2 activation, mitochondrial damage followed by downstream caspase 9 and -3 activation, and cell apoptosis in human lung epithelial cell line A549. When A549 cells were pretreated with the caspase-2 inhibitor benzyloxycarbonyl Val-Asp(-OMe)-Val-Ala-Asp(-OMe)-fluoromethyl ketone or transfected with caspase-2 short interfering RNA, both ceramide- and etoposide-induced mitochondrial damage and apoptosis were blocked. Overexpression of Bcl-2 prevented ceramide- and etoposide-induced caspase-2 activation and mitochondrial apoptosis. Furthermore, caspase-2 was activated when A549 cells were introduced with Bcl-2 short interfering RNA or were treated with Bcl-2 inhibitor, which provided direct evidence of a negative regulatory effect of Bcl-2 on caspase-2. Cell survival was observed when caspase-2 was inhibited in Bcl-2-silencing cells. Blockage of the mitochondrial permeability transition pore and caspase-9 demonstrated that Bcl-2 modulated caspase-2 activity occurred upstream of mitochondria. Further studies showed that Bcl-2 was dephosphorylated at serine 70 after ceramide and etoposide treatment. A protein phosphatase inhibitor, okadaic acid, rescued Bcl-2 dephosphorylation and blocked caspase-2 activation, mitochondrial damage, and cell death. Taken together, ceramide and etoposide induced mitochondria-mediated apoptosis by initiating caspase-2 activation, which was, at least in part, regulated by Bcl-2. PMID- 15817480 TI - Characterization of a novel positive transcription regulatory element that differentially regulates the insulin-like growth factor binding protein-3 (IGFBP 3) gene in senescent cells. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is a well documented growth inhibitor and pro-apoptotic factor. IGFBP-3 mRNA and its protein are overexpressed by senescent human diploid fibroblasts. However, the mechanism responsible for the up-regulation of its expression is still unclear. This report describes a novel transcriptional regulatory element, IGFBP-3 enhancer element (IEE), identified in the 5' untranslated region of the IGFBP-3 gene. This element differentially activates IGFBP-3 expression in senescent versus young fibroblasts. Electrophoretic mobility shift assays revealed abundant complexes in senescent cell nuclear extracts compared with young cell nuclear extracts. Similar to young proliferative cells, young quiescent cells showed reduced binding activity; enhancement of this activity was specific to senescent cells and not an effect of cell cycle arrest. The DNase I footprint revealed the protein-binding core sequence within the IEE through which the protein binds the IEE. Site-directed mutagenesis within IEE abolished binding activity and selectively decreased IGFBP-3 promoter activity in senescent (but not young) cells. Furthermore, introduction of an IEE decoy suppressed the endogenous IGFBP 3 gene expression specifically in senescent cells. These results point to the IEE as being a positive transcription regulatory element that contributes to the up regulation of IGFBP-3 during cellular senescence. PMID- 15817481 TI - Modeling and experimental validation of the binary complex of the plectin actin binding domain and the first pair of fibronectin type III (FNIII) domains of the beta4 integrin. AB - The binding of plectin to the beta4 subunit of the alpha6beta4 integrin is a critical step in the formation of hemidesmosomes. An important interaction between these two proteins occurs between the actin-binding domain (ABD) of plectin and the first pair of fibronectin type III (FNIII) domains and a small part of the connecting segment of beta4. Previously, a few amino acids, critical for this interaction, were identified in both plectin and beta4 and mapped on the crystal structures of the ABD of plectin and the first pair of FNIII domains of beta4. In the present study, we used this biochemical information and protein protein docking calculations to construct a model of the binary complex between these two protein domains. The top scoring computational model predicts that the calponin-homology 1 (CH1) domain of the ABD associates with the first and the second FNIII domains of beta4. Our mutational analysis of the residues at the proposed interface of both the FNIII and the CH1 domains is in agreement with the suggested interaction model. Computational simulations to predict protein motions suggest that the exact model of FNIII and plectin CH1 interaction might well differ in detail from the suggested model due to the conformational plasticity of the FNIII domains, which might lead to a closely related but different mode of interaction with the plectin-ABD. Furthermore, we show that Ser-1325 in the connecting segment of beta4 appears to be essential for the recruitment of plectin into hemidesmosomes in vivo. This is consistent with the proposed model and previously published mutational data. In conclusion, our data support a model in which the CH1 domain of the plectin-ABD associates with the groove between the two FNIII domains of beta4. PMID- 15817482 TI - Expression of bovine F1-ATPase with functional complementation in yeast Saccharomyces cerevisiae. AB - The mitochondrial F(1)F(0)-ATP synthase is a multimeric enzyme complex composed of at least 16 unique peptides with an overall molecular mass of approximately 600 kDa. F(1)-ATPase is composed of alpha(3)beta(3)gammadeltaepsilon with an overall molecular mass of 370 kDa. The genes encoding bovine F(1)-ATPase have been expressed in a quintuple yeast Saccharomyces cerevisiae deletion mutant (DeltaalphaDeltabetaDeltagammaDeltadeltaDeltaepsilon). This strain expressing bovine F(1) is unable to grow on medium containing a non-fermentable carbon source (YPG), indicating that the enzyme is non-functional. However, daughter strains were easily selected for growth on YPG medium and these were evolved for improved growth on YPG medium. The evolution of the strains was presumably due to mutations, but mutations in the genes encoding the subunits of the bovine F(1) ATPase were not required for the ability of the cell to grow on YPG medium. The bovine enzyme expressed in yeast was partially purified to a specific activity of about half of that of the enzyme purified from bovine heart mitochondria. These results indicate that the molecular machinery required for the assembly of the mitochondrial ATP synthase is conserved from bovine and yeast and suggest that yeast may be useful for the expression, mutagenesis, and analysis of the mammalian F(1)- or F(1)F(0)-ATP synthase. PMID- 15817483 TI - PYPAF3, a PYRIN-containing APAF-1-like protein, is a feedback regulator of caspase-1-dependent interleukin-1beta secretion. AB - PYPAF3 is a member of the PYRIN-containing apoptotic protease-activating factor-1 like proteins (PYPAFs, also called NALPs). Among the members of this family, PYPAF1, PYPAF5, PYPAF7, and NALP1 have been shown to induce caspase-1-dependent interleukin-1beta secretion and NF-kappaB activation in the presence of the adaptor molecule ASC. On the other hand, we recently discovered that PYNOD, another member of this family, is a suppressor of these responses. Here, we show that PYPAF3 is the second member that inhibits caspase-1-dependent interleukin 1beta secretion. In contrast, PYPAF2/NALP2 does not inhibit this response but rather inhibits the NF-kappaB activation that is induced by the combined expression of PYPAF1 and ASC. Both PYPAF2 and PYPAF3 mRNAs are broadly expressed in a variety of tissues; however, neither is expressed in skeletal muscle, and only PYPAF2 mRNA is expressed in heart and brain. They are also expressed in many cell lines of both hematopoietic and non-hematopoietic lineages. Stimulation of monocytic THP-1 cells with lipopolysaccharide or interleukin-1beta induced PYPAF3 mRNA expression. Furthermore, the stable expression of PYPAF3 in THP-1 cells abrogated the ability of the cells to produce interleukin-1beta in response to lipopolysaccharide. These results suggest that PYPAF3 is a feedback regulator of interleukin-1beta secretion. Thus, PYPAF2 and PYPAF3, together with PYNOD, constitute an anti-inflammatory subgroup of PYPAFs. PMID- 15817484 TI - Probing the beta2 adrenoceptor binding site with catechol reveals differences in binding and activation by agonists and partial agonists. AB - The beta(2) adrenergic receptor (beta(2)AR) is a prototypical family A G protein coupled receptor (GPCR) and an excellent model system for studying the mechanism of GPCR activation. The beta(2)AR agonist binding site is well characterized, and there is a wealth of structurally related ligands with functionally diverse properties. In the present study, we use catechol (1,2-benzenediol, a structural component of catecholamine agonists) as a molecular probe to identify mechanistic differences between beta(2)AR activation by catecholamine agonists, such as isoproterenol, and by the structurally related non-catechol partial agonist salbutamol. Using biophysical and pharmacologic approaches, we show that the aromatic ring of salbutamol binds to a different site on the beta(2)AR than the aromatic ring of catecholamines. This difference is important in receptor activation as it has been hypothesized that the aromatic ring of catecholamines plays a role in triggering receptor activation through interactions with a conserved cluster of aromatic residues in the sixth transmembrane segment by a rotamer toggle switch mechanism. Our experiments indicate that the aromatic ring of salbutamol does not activate this mechanism either directly or indirectly. Moreover, the non-catechol ring of partial agonists does not interact optimally with serine residues in the fifth transmembrane helix that have been shown to play an important role in activation by catecholamines. These results demonstrate unexpected differences in binding and activation by structurally similar agonists and partial agonists. Moreover, they provide evidence that activation of a GPCR is a multistep process that can be dissected into its component parts using agonist fragments. PMID- 15817485 TI - Identification of a subversive substrate of Trichomonas vaginalis purine nucleoside phosphorylase and the crystal structure of the enzyme-substrate complex. AB - Trichomonas vaginalis is an anaerobic protozoan parasite that causes trichomoniasis, a common sexually transmitted disease with worldwide impact. One of the pivotal enzymes in its purine salvage pathway, purine nucleoside phosphorylase (PNP), shows physical properties and substrate specificities similar to those of the high molecular mass bacterial PNPs but differing from those of human PNP. While carrying out studies to identify inhibitors of T. vaginalis PNP (TvPNP), we discovered that the nontoxic nucleoside analogue 2 fluoro-2'-deoxyadenosine (F-dAdo) is a "subversive substrate." Phosphorolysis by TvPNP of F-dAdo, which is not a substrate for human PNP, releases highly cytotoxic 2-fluoroadenine (F-Ade). In vitro studies showed that both F-dAdo and F Ade exert strong inhibition of T. vaginalis growth with estimated IC(50) values of 106 and 84 nm, respectively, suggesting that F-dAdo might be useful as a potential chemotherapeutic agent against T. vaginalis. To understand the basis of TvPNP specificity, the structures of TvPNP complexed with F-dAdo, 2 fluoroadenosine, formycin A, adenosine, inosine, or 2'-deoxyinosine were determined by x-ray crystallography with resolutions ranging from 2.4 to 2.9 A. These studies showed that the quaternary structure, monomer fold, and active site are similar to those of Escherichia coli PNP. The principal active site difference is at Thr-156, which is alanine in E. coli PNP. In the complex of TvPNP with F-dAdo, Thr-156 causes the purine base to tilt and shift by 0.5 A as compared with the binding scheme of F-dAdo in E. coli PNP. The structures of the TvPNP complexes suggest opportunities for further improved subversive substrates beyond F-dAdo. PMID- 15817486 TI - The calpain system is involved in the constitutive regulation of beta-catenin signaling functions. AB - Beta-catenin is a multifunctional protein serving both as a structural element in cell adhesion and as a signaling component in the Wnt pathway, regulating embryogenesis and tumorigenesis. The signaling fraction of beta-catenin is tightly controlled by the adenomatous polyposis coli-axin-glycogen synthase kinase 3beta complex, which targets it for proteasomal degradation. It has been recently shown that Ca(2+) release from internal stores results in nuclear export and calpain-mediated degradation of beta-catenin in the cytoplasm. Here we have highlighted the critical relevance of constitutive calpain pathway in the control of beta-catenin levels and functions, showing that small interference RNA knock down of endogenous calpain per se (i.e. in the absence of external stimuli) induces an increase in the free transcriptional competent pool of endogenous beta catenin. We further characterized the role of the known calpain inhibitors, Gas2 and Calpastatin, demonstrating that they can also control levels, function, and localization of beta-catenin through endogenous calpain regulation. Finally we present Gas2 dominant negative (Gas2DN) as a new tool for regulating calpain activity, providing evidence that it counteracts the described effects of both Gas2 and Calpastatin on beta-catenin and that it works via calpain independently of the classical glycogen synthase kinase 3beta and proteasome pathway. Moreover, we provide in vitro biochemical evidence showing that Gas2DN can increase the activity of calpain and that in vivo it can induce degradation of stabilized/mutated beta-catenin. In fact, in a context where the classical proteasome pathway is impaired, as in colon cancer cells, Gas2DN biological effects accounted for a significant reduction in proliferation and anchorage independent growth of colon cancer. PMID- 15817487 TI - Evidence that interspecies polymorphism in the human and rat cholecystokinin receptor-2 affects structure of the binding site for the endogenous agonist cholecystokinin. AB - The cholecystokinin (CCK) receptor-2 exerts very important central and peripheral functions by binding the neuropeptides cholecystokinin or gastrin. Because this receptor is a potential therapeutic target, great interest has been devoted to the identification of efficient antagonists. However, interspecies genetic polymorphism that does not alter cholecystokinin-induced signaling was shown to markedly affect activity of synthetic ligands. In this context, precise structural study of the agonist binding site on the human cholecystokinin receptor-2 is a prerequisite to elucidating the molecular basis for its activation and to optimizing properties of synthetic ligands. In this study, using site-directed mutagenesis and molecular modeling, we delineated the binding site for CCK on the human cholecystokinin receptor-2 by mutating amino acids corresponding to that of the rat homolog. By doing so, we demonstrated that, although resembling that of rat homolog, the human cholecystokinin receptor-2 binding site also displays important distinct structural features that were demonstrated by susceptibility to several point mutations (F120A, Y189A, H207A). Furthermore, docking of CCK in the human and rat cholecystokinin receptor-2, followed by dynamic simulations, allowed us to propose a plausible structural explanation of the experimentally observed difference between rat and human cholecystokinin-2 receptors. PMID- 15817488 TI - Post-transcriptional regulation of the yeast high affinity iron transport system. AB - Saccharomyces cerevisiae transcriptionally regulates the expression of the plasma membrane high affinity iron transport system in response to iron need. This transport system is comprised of the products of the FET3 and FTR1 genes. We show that Fet3p and Ftr1p are post-translationally regulated by iron. Incubation of cells in high iron leads to the internalization and degradation of both Fet3p and Ftr1p. Yeast strains defective in endocytosis (Deltaend4) show a reduced iron induced loss of Fet3p-Ftr1p. In cells with a deletion in the vacuolar protease PEP4, high iron medium leads to the accumulation of Fet3p and Ftr1p in the vacuole. Iron-induced degradation of Fet3p-Ftr1p is significantly reduced in strains containing a deletion of a gene, VTA1, which is involved in multivesicular body (MVB) sorting in yeast. Sorting through the MVB can involve ubiquitination. We demonstrate that Ftr1p is ubiquitinated, whereas Fet3p is not ubiquitinated. Iron-induced internalization and degradation of Fet3p-Ftr1p occurs in a mutant strain of the E3 ubiquitin ligase RSP5 (rsp5-1), suggesting that Rsp5p is not required. Internalization of Fet3p-Ftr1p is specific for iron and requires both an active Fet3p and Ftr1p, indicating that it is the transport of iron through the iron permease Ftr1p that is responsible for the internalization and degradation of the Fet3p-Ftr1p complex. PMID- 15817489 TI - Identification of the minimal domain structure of bone morphogenetic protein-1 (BMP-1) for chordinase activity: chordinase activity is not enhanced by procollagen C-proteinase enhancer-1 (PCPE-1). AB - Bone morphogenetic protein 1 (BMP-1), which is a tolloid member of the astacin like family of zinc metalloproteinases, is a highly effective procollagen C proteinase (PCP) and chordinase. On the other hand, mammalian tolloid like-2 (mTLL-2) does not cleave chordin or procollagen; procollagen is cleaved by mTLL-2 in the presence of high levels of procollagen C-proteinase enhancer-1 (PCPE-1), for reasons that are unknown. We used these differences in activity between BMP-1 and mTLL-2 to narrow in on the domains in BMP-1 that specify PCP and chordinase activity. Using a domain swap approach, we showed that: 1) the metalloproteinase and CUB2 domains of BMP-1 are absolutely required for PCP activity; swaps with either of the corresponding domains in BMP-1 and mTLL-2 did not result in procollagen cleavage and 2) the proteinase domain of mTLL-2 can cleave chordin if coupled to the CUB1 domain of BMP-1. Therefore, the minimal structure for chordinase activity comprises a metalloproteinase domain (either from BMP-1 or from mTLL-2) and the CUB1 domain of BMP-1 (the CUB1 domain of mTLL-2 cannot substitute for the CUB1 domain of BMP-1). We showed that the minimal procollagen C-proteinase (BMP-1 lacking the EGF and CUB3 domain) was enhanced by PCPE-1 but not as well as BMP-1 retaining the CUB3 domain. Further studies showed that PCPE 1 had no effect on the ability of BMP-1 to cleave chordin. The data support a previously suggested mechanism of PCPE-1 whereby PCPE-1 interacts with procollagen, but in addition, the CUB3 domain of BMP-1 appears to augment the interaction. PMID- 15817490 TI - Regulation of phospholipase C-delta1 through direct interactions with the small GTPase Ral and calmodulin. AB - Second messengers generated from membrane lipids play a critical role in signaling and control diverse cellular processes. Despite being one of the most evolutionarily conserved of all the phosphoinositide-specific phospholipase C (PLC) isoforms, a family of enzymes responsible for hydrolysis of the membrane lipid phosphatidylinositol bisphosphate, the mechanism of PLC-delta1 activation is still poorly understood. Here we report a novel regulatory mechanism for PLC delta1 activation that involves direct interaction of the small GTPase Ral and the universal calcium-signaling molecule calmodulin (CaM) with PLC-delta1. In addition, we have identified a novel IQ type CaM binding motif within the catalytic region of PLC-delta1 that is not found in other PLC isoforms. Binding of CaM at the IQ motif inhibits PLC-delta1 activity, while addition of Ral reverses the inhibition. The overexpression of various Ral mutants in cells potentiates PLC-delta1 activity. Thus, the Ral-CaM complex defines a multifaceted regulatory mechanism for PLC-delta1 activation. PMID- 15817492 TI - Identification of pressure passive cerebral perfusion and its mediators after infant cardiac surgery. PMID- 15817491 TI - Defining a minimal motif required to prevent connexin oligomerization in the endoplasmic reticulum. AB - In contrast to most multimeric transmembrane complexes that oligomerize in the endoplasmic reticulum (ER), the gap junction protein connexin43 (Cx43) oligomerizes in an aspect of the Golgi apparatus. The mechanisms that prevent oligomerization of Cx43 and related connexins in the ER are not well understood. Also, some studies suggest that connexins can oligomerize in the ER. We used connexin constructs containing a C-terminal dilysine-based ER retention/retrieval signal (HKKSL) transfected into HeLa cells to study early events in connexin oligomerization. Using this approach, Cx43-HKKSL was retained in the ER and prevented from oligomerization. However, another ER-retained HKKSL-tagged connexin, Cx32-HKKSL, had the capacity to oligomerize. Because this suggested that Cx43 contains a motif that prevented oligomerization in the ER, a series of HKKSL-tagged and untagged Cx32/Cx43 chimeras was screened to define this motif. The minimal motif, which prevented ER oligomerization, consisted of the complete third transmembrane domain and the second extracellular loop from Cx43 on a Cx32 backbone. We propose that charged residues present in Cx43 and related connexins help prevent ER oligomerization by stabilizing the third transmembrane domain in the membrane bilayer. PMID- 15817493 TI - Controversial nutrients that potentially affect preterm neurodevelopment: essential fatty acids and iron. PMID- 15817494 TI - Endothelin-1 in congenital heart disease. AB - Endothelin-1 (ET-1) is a 21-amino acid polypeptide produced primarily by vascular endothelial cells. First discovered in 1988 as a potent vasoconstrictor, it has subsequently been appreciated to participate in several biologic activities, including vascular smooth muscle proliferation, fibrosis, cardiac and vascular hypertrophy, and inflammation. Increasing data demonstrate alterations in ET-1 signaling in newborns, infants, and children with congenital heart defects that are associated with alterations in pulmonary blood flow. This review outlines the pathophysiologic role of the ET-1 cascade in the development of altered pulmonary vascular tone and reactivity that occurs with congenital heart disease and its repair, following the use of cardiopulmonary bypass. In addition, therapeutic implications for the use of novel ET receptor antagonists will be emphasized. PMID- 15817495 TI - NPHS2 (Podocin) mutations in nephrotic syndrome. Clinical spectrum and fine mechanisms. AB - Nephrotic syndrome (NS) is the most frequent cause of proteinuria in children and is emerging as a leading cause of uremia. Molecular studies in families with recessive NS have led to the discovery of specialized molecules endowed in podocytes that play a role in proteinuria. This review focalizes the key position of podocin (NPHS2 gene) in this rapidly evolving field and furnishes a compendium to those involved in clinics and genetics of NS. Screening for NPHS2 mutations have been done in sporadic NS and familial cases with recessive inheritance, documenting a mutation detection rate of 45-55% in families and 8-20% in sporadic NS according to the different groups and considering all the clinical phenotypes. Almost 50 NPHS2 mutations have been reported and variants and/or non silent polymorphisms potentially involved in proteinuria were recognized. Personalized data on clinical aspects related to responsiveness to drugs, evolution to end stage renal failure and post-transplant outcome are reported. Functional studies and cell sorting experiments demonstrated retention in the endoplasmic reticulum of most mutants involving the stomatin domain. Pull-down experiments with the common R229Q polymorphism demonstrated an altered interaction with nephrin that affects the stability of the functional unit. Overall, data are here presented that underscore a major role of inherited defects of NPHS2 in NS in children (including a relevant impact in sporadic cases) and give the functional rationale for the association. A practical compendium is also given to clinicians involved in the management of NS that should modify the classic therapeutic approach. PMID- 15817496 TI - Protein and energy requirements of the preterm/low birthweight (LBW) infant. PMID- 15817497 TI - Notch signaling, brain development, and human disease. AB - The Notch signaling pathway is central to a wide array of developmental processes in a number of organ systems, including hematopoiesis, somitogenesis, vasculogenesis, and neurogenesis. These processes involve maintenance of stem cell self-renewal, proliferation, specification of cell fate or differentiation, and apoptosis. Recent studies have led to the recognition of the role of the Notch pathway in early neurodevelopment, learning, and memory, as well as late life neurodegeneration. This review summarizes what is currently known about the role of the Notch pathway in neural stem cells, gliogenesis, learning and memory, and neurologic disease. PMID- 15817499 TI - Control mechanisms of lung alveolar development and their disorders in bronchopulmonary dysplasia. AB - Bronchopulmonary dysplasia (BPD) is a chronic lung disease that occurs in very premature infants and is characterized by impaired alveologenesis. This ultimate phase of lung development is mostly postnatal and allows growth of gas-exchange surface area to meet the needs of the organism. Alveologenesis is a highly integrated process that implies cooperative interactions between interstitial, epithelial, and vascular compartments of the lung. Understanding of its underlying mechanisms has considerably progressed recently with identification of structural, signaling, or remodeling molecules that are crucial in the process. Thus, the pivotal role of elastin deposition in lung walls has been demonstrated, and many key control-molecules have been identified, including various transcription factors, growth factors such as platelet-derived growth factor, fibroblast growth factors, and vascular endothelial growth factor, matrix remodeling enzymes, and retinoids. BPD-associated changes in lung expression/content have been evidenced for most of these molecules, especially for signaling pathways, through both clinical investigations in premature infants and the use of animal models, including the premature baboon or lamb, neonatal exposure to hyperoxia in rodents, and maternal-fetal infection. These findings open therapeutic perspectives to correct imbalanced signaling. Unraveling the intimate molecular mechanisms of alveolar building appears as a prerequisite to define new strategies for the prevention and care of BPD. PMID- 15817498 TI - Fetal fatty acid oxidation disorders, their effect on maternal health and neonatal outcome: impact of expanded newborn screening on their diagnosis and management. AB - Mitochondrial fatty acid oxidation disorders (FAOD) are recessively inherited errors of metabolism. Newborns with FAOD typically present with hypoketotic hypoglycemia, metabolic acidosis, hepatic failure, and cardiomyopathy. Late presentations include episodic myopathy, neuropathy, retinopathy, and arrhythmias. Sudden unexpected death can occur at any age and can be confused with sudden infant death syndrome. Some FAOD are associated with intrauterine growth restriction, prematurity, and pregnancy complications in the heterozygous mother, such as severe preeclampsia, acute fatty liver of pregnancy (AFLP), or hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome. Maternal pregnancy complications occur primarily in mothers carrying a fetus with long chain l-3-hydroxyacyl CoA dehydrogenase deficiency or general trifunctional protein deficiencies. FAOD as a group represent the most common inborn errors of metabolism, and presymptomatic diagnosis of FAOD is the key to reduce morbidity and avoid mortality. The application of tandem mass spectrometry to newborn screening provides an effective means to identify most FAOD patients presymptomatically. At the beginning of 2005, 36 state newborn screening programs have mandated or adopted this technology resulting in a marked increase in the number of asymptomatic neonates with FAOD diagnosed. To ensure the long-term benefits of such screening programs, pediatricians and other health care providers must be educated about these disorders and their treatment. PMID- 15817500 TI - Developmental expression of vasoactive and growth factors in human lung. Role in pulmonary vascular resistance adaptation at birth. AB - The factors that mediate the postnatal fall in pulmonary vascular resistance, which is crucial for normal gas exchange, are not fully understood. The endothelium has been implicated in this phenomenon, through the release of vasorelaxant factors such as nitric oxide (NO). Human pulmonary expression of endothelial NO synthase increases up to 31 wk of gestation, together with vascular endothelial growth factor (VEGF), and both factors potently mediate pulmonary angiogenesis and vasorelaxation. During the perinatal period, when pulmonary vasodilatation is maximal, endothelial NO synthase and VEGF are weakly expressed. This raises the involvement of vasorelaxant factors other than NO at birth. One candidate is endothelial-derived hyperpolarizing factor, which induces smooth muscle cell hyperpolarization by activating K(ATP) channels. The marked vasorelaxation induced by activation of these channels in newborn animals, and their strong perinatal expression in the human lung, suggest their involvement during this phase. Another candidate is endothelin (ET)-1, together with its receptors ET-A and ET-B. ET-A receptors are located exclusively on smooth muscle cells and mediate vasoconstriction, whereas ET-B receptors mediate vasoconstriction when located on smooth muscle cells and vasodilatation when located on endothelial cells. ET-B receptors, which are strongly expressed in the human fetal lung both at the end of gestation and after birth, may be involved in perinatal pulmonary vasodilatation. Thus, in human fetal lung, K(ATP) channels and ET-B receptors could be important in mediating the perinatal pulmonary vasodilatation crucial for adapting the pulmonary circulation to extrauterine life. PMID- 15817501 TI - Genetic approaches to studying common diseases and complex traits. AB - Most common diseases and most quantitative traits that can be measured in human populations are complex genetic traits. That is, many genetic and nongenetic factors interact to determine the final phenotype, whether that phenotype is susceptibility to disease, or a quantifiable trait such as height, weight, serum cholesterol, or blood pressure. Identifying the genes that underlie the population variation in these phenotypes has been challenging. Recently, databases of common genetic variants, recognition of the patterns of genetic variation, and rapid genotyping methodologies have emerged, and the combination of these tools and resources will greatly facilitate genetic association studies, a potentially powerful method to map the genes for complex traits. However, care will be required in performing and interpreting these association studies. Until genome-wide studies are feasible, choosing candidate genes will be necessary. In addition, the choice of phenotype will likely influence the success of these gene mapping efforts. Finally, population genetic methods, including searching for genes under selection, may provide clues to the location of the genes for common disease and complex traits. PMID- 15817502 TI - New members of the insulin family: regulators of metabolism, growth and now ... reproduction. AB - Insulin, IGF, and relaxin are established members of the insulin protein superfamily. The application of the techniques of cellular, molecular, and computational biology has permitted the identification of new insulin-like ligands and their cognate receptors. Information regarding the biologic role is available for some of these newly identified ligand-receptor systems and indicates novel roles in diverse processes such as testicular descent, germ cell function, and cell migration. PMID- 15817503 TI - Water channels (aquaporins) and their role for postnatal adaptation. AB - Birth is a transition from an underwater life in the uterus to a terrestrial life in a milieu where supply of water is limited. Rapid adaptation to the new environment is crucial for survival and health of infants. The discovery of a family of molecules-aquaporin (AQP) water channels-that are responsible for regulated water transport across cell membranes has made it possible to identify the molecular mechanisms behind the postnatal homeostatic adaptation and to better understand water imbalance-related disorders in infancy and childhood. Thirteen mammalian AQP isoforms have been identified, most of them having a unique tissue-specific pattern of expression. Most mammalian AQPs can be dynamically regulated, which makes them potential targets for the development of new drugs for diseases associated with disturbances in water homeostasis. This review deals with AQP in kidney, lung, and brain. Evidence is presented that AQPs are expressed in a specific age-dependent manner and that the timed expression of AQPs may have a crucial role during the early postnatal period. PMID- 15817504 TI - Potential for protection and repair following injury to the developing brain: a role for erythropoietin? AB - Perinatal brain injury is a major contributor to perinatal morbidity and mortality, and a considerable number of these children will develop long term neurodevelopmental disabilities. Despite the severe clinical and socio-economic significance and the advances in neonatal care over the past twenty years, no therapy yet exists that effectively prevents or ameliorates detrimental neurodevelopmental effects in cases of perinatal/neonatal brain injury. Our objective is to review recent evidence in relation to the pervading hypothesis for targeting time-dependent molecular and cellular repair mechanisms in the developing brain. In addition we review several potential neuroprotective strategies specific to the developing nervous system, with a focus on erythropoietin (Epo) because of its potential role in protection as well as repair. PMID- 15817505 TI - Molecular mechanisms of early lung specification and branching morphogenesis. AB - The "hard wiring" encoded within the genome that determines the emergence of the laryngotracheal groove and subsequently early lung branching morphogenesis is mediated by finely regulated, interactive growth factor signaling mechanisms that determine the automaticity of branching, interbranch length, stereotypy of branching, left-right asymmetry, and finally gas diffusion surface area. The extracellular matrix is an important regulator as well as a target for growth factor signaling in lung branching morphogenesis and alveolarization. Coordination not only of epithelial but also endothelial branching morphogenesis determines bronchial branching and the eventual alveolar-capillary interface. Improved prospects for lung protection, repair, regeneration, and engineering will depend on more detailed understanding of these processes. Herein, we concisely review the functionally integrated morphogenetic signaling network comprising the critical bone morphogenetic protein, fibroblast growth factor, Sonic hedgehog, transforming growth factor-beta, vascular endothelial growth factor, and Wnt signaling pathways that specify and drive early embryonic lung morphogenesis. PMID- 15817506 TI - Unraveling the pathogenesis and etiology of biliary atresia. AB - Biliary atresia (BA) is the most common and important neonatal hepatobiliary disorder. Because current treatment is inadequate, there is an urgent need to better understand the etiology and pathogenesis of BA. Two major forms of BA are recognized: an embryonic form associated with other congenital anomalies and a perinatal form in which bile ducts were presumably formed normally but underwent fibro-obliteration in the perinatal period. There are currently several proposed pathogenic pathways leading to the phenotype of BA, including an immune or autoimmune response to a perinatal insult (e.g. cholangiotropic viral infection) and dysregulated embryonic development of the extra- or intrahepatic biliary system. Recent advances in developmental biology, genomics and genetics, and cell immunology and biology, coupled with the development of appropriate animal models, have provided support for these postulated mechanisms. Future investigations combining animal model work and evaluation of clinical specimens holds the promise of identifying the etiology of BA and providing a scientific basis for treatment and preventative interventions. PMID- 15817507 TI - Molecular basis of adrenal insufficiency. AB - Defective production of adrenal steroids due to either primary adrenal failure or hypothalamic-pituitary impairment of the corticotrophic axis causes adrenal insufficiency. Depending on the etiologies of adrenal insufficiency, clinical manifestations may be severe or mild, have gradual or sudden onset, begin in infancy or childhood/adolescence. Adrenal crisis represents an endocrine emergency, and thus the rapid recognition and prompt therapy for adrenal crisis are critical for survival even before the diagnosis is made. The recognition of various disorders that cause adrenal insufficiency, either at a clinical or molecular level, often has implications for the management of the patient. Recent molecular-genetic analysis for the disorder that causes adrenal insufficiency gives valuable insights into the adrenal organogenesis, the regulation of steroid hormone biosynthesis, and the developmental and reproductive endocrinology. In this review we present the latest information on the molecular basis of adrenal insufficiency, with special emphasis on congenital lipoid adrenal hyperplasia, P450-oxidoreductase deficiency, and adrenal hypoplasia congenita. PMID- 15817508 TI - Soluble Fms-like tyrosine kinase 1 and endothelial dysfunction in the pathogenesis of preeclampsia. AB - Preeclampsia, a pregnancy-specific syndrome of hypertension and proteinuria, is characterized by defective placental vasculogenesis and widespread maternal endothelial dysfunction. Although the manifestations of preeclampsia are primarily maternal, the burden of morbidity and mortality is often on the neonate, since the only effective treatment-delivery of the fetus and placenta often results in iatrogenic prematurity. In this review, we summarize recent advances in our understanding of the pathophysiology of preeclampsia, including normal and aberrant placental vascular development and evidence for endothelial dysfunction. We describe recent evidence that supports a novel mechanism in which a maladaptive shift in placental production of angiogenic factors such as soluble fms-like tyrosine kinase 1 (a circulating antiangiogenic protein) may play an important role in the pathogenesis of preeclampsia. PMID- 15817509 TI - Nuclear envelopathies--raising the nuclear veil. AB - The nuclear envelope separates the chromosomes from cytoplasm in eukaryotic cells and consists of three main domains: inner and outer nuclear membranes and nuclear pore complexes. The inner nuclear membrane maintains close associations with the underlying chromatin and nuclear lamina. For many years, the nuclear envelope was thought to function mainly as an architectural stabilizer of the nucleus, participating in assembly and disassembly processes during mitosis. However, recent findings demonstrate that nuclear envelope proteins are involved in fundamental nuclear functions, such as gene transcription and DNA replication, and that inherited or de novo mutated proteins cause human diseases, termed "nuclear envelopathies." These findings emphasize the importance of understanding the functions of this cellular domain, in both physiologic and pathologic states. To date, mutations in the genes encoding the nuclear envelope proteins emerin, MAN1, lamin A/C, and lamin B receptor were found to cause nuclear envelopathies. The diseases that are caused by mutations in LMNA gene are collectively called "laminopathies." Nuclear envelopathies have diverse clinical phenotypes, ranging from cardiac and skeletal myopathies to partial lipodystrophy, peripheral neuropathy, and premature aging. This raises the question of how do such ubiquitously expressed proteins give rise to tissue-specific disease phenotypes. One possible explanation is the involvement of nuclear envelope proteins in the regulation of gene transcription, a novel mechanism that has been the focus of research in our lab in recent years. In this review, we describe recent discoveries in the field of nuclear envelopathies and discuss current proposed pathophysiological mechanisms underlying these diseases. PMID- 15817510 TI - Somatosensory activations during the observation of touch and a case of vision touch synaesthesia. AB - In this study, we describe a new form of synaesthesia in which visual perception of touch elicits conscious tactile experiences in the perceiver. We describe a female subject (C) for whom the observation of another person being touched is experienced as tactile stimulation on the equivalent part of C's own body. Apart from this clearly abnormal synesthetic experience, C is healthy and normal in every other way. In this study, we investigate whether C's 'mirrored touch' synesthetic experience is caused by overactivity in the neural system that responds to the observation of touch. A functional MRI experiment was designed to investigate the neural system involved in the perception of touch in a group of 12 non-synesthetic control subjects and in C. We investigated neural activity to the observation of touch to a human face or neck compared with the observation of touch to equivalent regions on an object. Furthermore, to investigate the somatosensory topography of the activations during observation of touch, we compared activations when observing a human face or neck being touched with activations when the subjects themselves were touched on their own face or neck. The results demonstrated that the somatosensory cortex was activated in the non synesthetic subjects by the mere observation of touch and that this activation was somatotopically organized such that observation of touch to the face activated the head area of primary somatosensory cortex, whereas observation of touch to the neck did not. Moreover, in non-synesthetic subjects, the brain's mirror system-comprising premotor cortex, superior temporal sulcus and parietal cortex-was activated by the observation of touch to another human more than to an object. C's activation patterns differed in three ways from those of the non synesthetic controls. First, activations in the somatosensory cortex were significantly higher in C when she observed touch. Secondly, an area in left premotor cortex was activated in C to a greater extent than in the non synesthetic group. Thirdly, the anterior insula cortex bilaterally was activated in C, but there was no evidence of such activation in the non-synesthetic group. The results suggest that, in C, the mirror system for touch is overactive, above the threshold for conscious tactile perception. PMID- 15817511 TI - Grey and white matter volume changes in early primary progressive multiple sclerosis: a longitudinal study. AB - We have recently reported brain atrophy in the early stages of primary progressive multiple sclerosis (PPMS), affecting both grey and white matter (GM and WM). However, to date no clinical or radiological predictors of GM and WM atrophy have been identified. The aim was to investigate short-term changes in GM and WM volumes and to assess the predictive value of demographic, clinical and radiological variables in order to gain a better understanding of the pathological substrate underlying these changes. Thirty-one subjects with PPMS within 5 years of symptom onset were studied at baseline and after 1 year. At baseline, patients underwent neurological examination and were scored on the Expanded Disability Status Scale (EDSS) and Multiple Sclerosis Functional Composite. They had 3D inversion-prepared fast spoiled gradient recalled (FSPGR), dual-echo and triple-dose post-contrast T1-weighted spin echo MRI scans. Proton density and enhancing lesion loads were determined. The 3DFSPGR sequence was repeated after 1 year and brain volume changes were calculated using two techniques, SPM99 (statistical parametric mapping) and SIENA (structural image evaluation, using normalization, of atrophy). Stepwise linear regression models were applied to baseline variables to identify independent predictors of atrophy development. Using SPM99, a decrease in brain parenchymal fraction (-1.03%; P < 0.001) and GM fraction (-1.49%; P < 0.001) was observed. The number of enhancing lesions independently predicted decrease in brain parenchymal fraction (P = 0.019) and decrease in WM fraction (P = 0.002). No independent predictors of GM fraction decrease were found. A mean brain volume change of -0.63% (range -4.27% to +1.18%; P = 0.002) was observed using SIENA, which was independently predicted by EDSS (P = 0.004). Global and GM atrophy can be detected over a 1-year period in early PPMS. The former may be predicted by the degree of inflammation, while the latter seems to be independent of it. SIENA and SPM-based methods appear to provide complementary information. PMID- 15817512 TI - The familial risk of subarachnoid haemorrhage. AB - Relatives of people with aneurysmal subarachnoid haemorrhage (SAH) may be at increased risk of SAH, but precise data on the level of risk and which relatives are most likely to be affected are lacking. We studied two samples: 5478 relatives of patients from the whole of Scotland who had a SAH in one year and 3213 relatives of patients with a SAH admitted to the West of Scotland regional neurosurgical unit 10 years previously. Overall, 2% of all relatives in each sample had a SAH. In the Scotland-wide sample, the absolute lifetime risk of SAH (from birth to 70 years) was higher for first-degree relatives [4.7%; 95% confidence interval (CI): 3.1-6.3%] than for second-degree (1.9%; 95% CI: 1.0 2.9%). In the West of Scotland sample, the lifetime risks were very similar to the Scotland-wide sample. The 10-year prospective risk for first-degree relatives alive at the time of the index patient's SAH was 1.2% (95% CI: 0.4-2%) and for second-degree was 0.5% (95% CI: 0.1-0.8%). There was a trend for risk to be highest in families with two first-degree relatives affected and lowest with only one second-degree affected. Most living relatives of patients who suffer a SAH are at low absolute risk of a future haemorrhage; screening is inappropriate except for the few families in whom two or more first-degree relatives, i.e. index case plus one extra have been affected. PMID- 15817513 TI - Functional MRI study of diencephalic amnesia in Wernicke-Korsakoff syndrome. AB - Anterograde amnesia in Wernicke-Korsakoff syndrome is associated with diencephalic lesions, mainly in the anterior thalamic nuclei. Whether diencephalic and temporal lobe amnesias are distinct entities is still not clear. We investigated episodic memory for faces using functional MRI (fMRI) in eight controls and in a 34-year-old man with Wernicke-Korsakoff syndrome and diencephalic lesions but without medial temporal lobe (MTL) involvement at MRI. fMRI was performed with a 1.5 tesla unit. Three dual-choice tasks were employed: (i) face encoding (18 faces were randomly presented three times and subjects were asked to memorize the faces); (ii) face perception (subjects indicated which of two faces matched a third face); and (iii) face recognition (subjects indicated which of two faces belonged to the group they had been asked to memorize during encoding). All activation was greater in the right hemisphere. In controls both the encoding and recognition tasks activated two hippocampal regions (anterior and posterior). The anterior hippocampal region was more activated during recognition. Activation in the prefrontal cortex was greater during recognition. In the subject with Wernicke-Korsakoff syndrome, fMRI did not show hippocampal activation during either encoding or recognition. During recognition, although behavioural data showed defective retrieval, the prefrontal regions were activated as in controls, except for the ventrolateral prefrontal cortex. fMRI activation of the visual cortices and the behavioural score on the perception task indicated that the subject with Wernicke-Korsakoff syndrome perceived the faces, paid attention to the task and demonstrated accurate judgement. In the subject with Wernicke-Korsakoff syndrome, although the anatomical damage does not involve the MTL, the hippocampal memory encoding has been lost, possibly as a consequence of the hippocampal-anterior thalamic axis involvement. Anterograde amnesia could therefore be the expression of damage to an extended hippocampal system, and the distinction between temporal lobe and diencephalic amnesia has limited value. In the subject with Wernicke-Korsakoff syndrome, the preserved dorsolateral prefrontal cortex activation during incorrect recognition suggests that this region is more involved in either the orientation or attention at retrieval than in retrieval. The lack of activation of the prefrontal ventrolateral cortex confirms the role of this area in episodic memory formation. PMID- 15817514 TI - Three to four years after diagnosis: cognition and behaviour in children with 'epilepsy only'. A prospective, controlled study. AB - A 3.5-year follow-up study of cognition and behaviour in 42 children with newly diagnosed idiopathic or cryptogenic epilepsy ('epilepsy only') attending mainstream education and 30 healthy gender-matched classmate controls was carried out to identify differences between groups, to detect factors that contribute to the difference and its change over time, and to establish the proportion of poorly performing children. The neuropsychological battery covered the major domains of cognition, mental and motor speed and academic language skills. Children were tested at the time of diagnosis (before any anti-epileptic drug treatment started) and 3, 12 and approximately 42 months later. Parents and teachers completed behaviour checklists, for which the scoring was adapted to prevent any influence of epilepsy-related ambiguity. Based on parental interviews at the time of diagnosis, children with epilepsy were categorized as having longstanding behavioural and/or learning problems, as belonging to a troubled family, as being exposed to 'off-balance' parenting starting at the time of epilepsy onset and/or as reacting maladaptively to the changes in relation to the onset of epilepsy. Throughout follow-up, the group of children with epilepsy only performed less well than healthy classmates on measures of learning, memory span for words, attention and behaviour. After controlling for school delay, proactive interference (number of responses to the same images as in the learning trials, but now presented in reordered locations) was the only remaining variable that distinguished the group of children with epilepsy only. Group-wise, no changes in cognitive and behavioural differences over time were found, but instability in individual performances appeared to characterize children with epilepsy only. Rather than intrinsically epilepsy-related variables, such as idiopathic versus cryptogenic aetiology, seizure control or anti-epileptic drug treatment, the child's prediagnostic learning and behavioural histories and the parents' ability to continue their habitual parenting in the face of the diagnosis of epilepsy only were shown by both group-wise and case-by-case analyses to be important for understanding the cognitive and behavioural functioning of the children with epilepsy only. PMID- 15817515 TI - Brain tissue damage in dementia with Lewy bodies: an in vivo diffusion tensor MRI study. AB - The aim of the present study was to apply diffusion tensor MRI (DT-MRI), a quantitative MRI measure which reflects tissue organization, to dementia with Lewy bodies (DLB). DT-MRI scans were obtained from 15 patients with probable DLB and 10 sex- and age-matched healthy controls. Abnormalities were found in the corpus callosum, pericallosal areas and the frontal, parietal, occipital and, less prominently, temporal white matter of patients compared with controls. Abnormalities were also found in the caudate nucleus and the putamen. The average grey matter volume was lower in patients than in controls. These findings of concomitant grey matter atrophy and white matter abnormalities (as detected by DT MRI) in regions with a high prevalence of long connecting fibre tracts might suggest the presence of neurodegeneration involving associative cortices. The modest involvement of the temporal lobe fits with the relative preservation of global neuropsychological measures and memory tasks in the early stage of DLB. The selective involvement of parietal, frontal and occipital lobes might explain some of the clinical and neuropsychological features of DLB, providing a possible distinctive marker for this disease. The abnormalities found in the subcortical grey matter may indicate that DLB and Parkinson's disease share a similar nigrostriatal involvement caused by common pathophysiological mechanisms. PMID- 15817516 TI - Lack of oestrogen protection in amyloid-mediated endothelial damage due to protein nitrotyrosination. AB - Amyloid beta-peptide (Abeta) cytotoxicity, the hallmark of Alzheimer's disease, implicates oxidative stress in both neurons and vascular cells, particularly endothelial cells. Consequently, antioxidants have shown neuroprotective activities against Abeta-induced cytotoxicity. Among the different antioxidants used in both in vitro and in vivo studies, 17beta-oestradiol (E2) has garnered the most attention. Oestrogen attenuated Abeta(E22Q)-induced toxicity in neurons but failed to protect endothelial cells. Here we show that E2-mediated activation of endothelial nitric oxide synthase (eNOS) increases the production of nitric oxide (NO), which, under Abeta(E22Q)-induced oxidative damage, results in the formation of peroxynitrite and increased nitration of tyrosine residues. Inhibition of eNOS prevents nitrotyrosination and permits E2-mediated protection against Abeta(E22Q) on endothelial cells. The main nitrotyrosinated proteins in the presence of E2 and Abeta(E22Q) were identified by MALDI-TOF mass spectrometry. These proteins are key players in the regulation of energy production, cytoskeletal integrity, protein metabolism and protection against oxidative stress. Our data highlight the potential damaging consequences of E2 in vascular disorders dealing with oxidative stress conditions, such as cerebral amyloid angiopathy, stroke and ischaemia-reperfusion conditions. PMID- 15817517 TI - Electrocorticographic high gamma activity versus electrical cortical stimulation mapping of naming. AB - Subdural electrocorticographic (ECoG) recordings in patients undergoing epilepsy surgery have shown that functional activation is associated with event-related broadband gamma activity in a higher frequency range (>70 Hz) than previously studied in human scalp EEG. To investigate the utility of this high gamma activity (HGA) for mapping language cortex, we compared its neuroanatomical distribution with functional maps derived from electrical cortical stimulation (ECS), which remains the gold standard for predicting functional impairment after surgery for epilepsy, tumours or vascular malformations. Thirteen patients had undergone subdural electrode implantation for the surgical management of intractable epilepsy. Subdural ECoG signals were recorded while each patient verbally named sequentially presented line drawings of objects, and estimates of event-related HGA (80-100 Hz) were made at each recording site. Routine clinical ECS mapping used a subset of the same naming stimuli at each cortical site. If ECS disrupted mouth-related motor function, i.e. if it affected the mouth, lips or tongue, naming could not be tested with ECS at the same cortical site. Because naming during ECoG involved these muscles of articulation, the sensitivity and specificity of ECoG HGA were estimated relative to both ECS-induced impairments of naming and ECS disruption of mouth-related motor function. When these estimates were made separately for 12 electrode sites per patient (the average number with significant HGA), the specificity of ECoG HGA with respect to ECS was 78% for naming and 81% for mouth-related motor function, and equivalent sensitivities were 38% and 46%, respectively. When ECS maps of naming and mouth related motor function were combined, the specificity and sensitivity of ECoG HGA with respect to ECS were 84% and 43%, respectively. This study indicates that event-related ECoG HGA during confrontation naming predicts ECS interference with naming and mouth-related motor function with good specificity but relatively low sensitivity. Its favourable specificity suggests that ECoG HGA can be used to construct a preliminary functional map that may help identify cortical sites of lower priority for ECS mapping. Passive recordings of ECoG gamma activity may be done simultaneously at all electrode sites without the risk of after-discharges associated with ECS mapping, which must be done sequentially at pairs of electrodes. We discuss the relative merits of these two functional mapping techniques. PMID- 15817518 TI - Fast modulation of heat-activated ionic current by proinflammatory interleukin 6 in rat sensory neurons. AB - The pro-inflammatory cytokine interleukin-6 (IL-6) together with its soluble receptor (sIL-6R) induces and maintains thermal hyperalgesia. It facilitates the heat-induced release of calcitonin gene-related peptide from rat cutaneous nociceptors in vivo and in vitro. Here we report that exposure of nociceptive neurons to the IL-6-sIL-6R complex or the gp130-stimulating designer IL-6-sIL-6R fusion protein Hyper-IL-6 (HIL-6) resulted in a potentiation of heat-activated inward currents (I(heat)) and a shift of activation thresholds towards lower temperatures without affecting intracellular calcium levels. The Janus tyrosine kinase inhibitor AG490, the selective protein kinase C (PKC) inhibitor, bisindolylmaleimide 1 (BIM1), as well as rottlerin, a selective blocker of the PKCdelta isoform, but not the cyclooxygenase inhibitor indomethacin, effectively reduced the effect. Reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization revealed expression of mRNA for the signal-transducing beta subunit of the receptor gp130 in neuronal somata, rather than satellite cells in rat dorsal root ganglia. Together, the results suggest that IL-6-sIL-6R acts directly on sensory neurons. It increases their susceptibility to noxious heat via the gp130/Jak/PKCdelta signalling pathway. PMID- 15817519 TI - Voxel-wise analysis of [123I]beta-CIT SPECT differentiates the Parkinson variant of multiple system atrophy from idiopathic Parkinson's disease. AB - To investigate the cerebral dopamine transporter status in the early stages of the parkinson-variant of multiple system atrophy (MSA-P), 15 patients with MSA-P and a disease duration up to 3 years were studied with [123I]beta-CIT single photon emission computed tomography (SPECT). Data were compared with 13 age matched healthy control subjects and 15 patients with idiopathic Parkinson's disease (IPD), matched for age and disease duration. Parametric SPECT images of the specific-to-nondisplaceable equilibrium partition coefficient (V3''), which is proportional to the receptor density (Bmax) have been generated. To objectively localize focal changes in dopaminergic function throughout the entire brain volume without having to make an a priori hypothesis as to their location, statistical parametric mapping (SPM) was applied to our [123I]beta-CIT SPECT study. Both MSA-P and IPD patients showed significant decreases in striatal [123I]beta-CIT SPECT uptake. However, in MSA-P patients an additional reduction in midbrain [123I]beta-CIT signal was localized with SPM compared with control subjects (MSA-P, V3'': 0.89 +/- 0.37 versus controls V3'': 1.81 +/- 0.38; P < 0.001) and patients with IPD (V3'': 1.84 +/- 0.26; P < 0.001). Stepwise linear discriminant analysis of mean [123I]beta-CIT uptake in the putamen, caudate and midbrain identified the caudate and midbrain as indices to classify correctly 95.2% of subjects as either normal, patients with MSA-P or IPD. Voxel-wise analysis of [123I]beta-CIT SPECT revealed more widespread decline of monoaminergic transporter availability in MSA-P compared with IPD, matching the underlying pathological features. We suggest that the quantification of midbrain DAT signal should be included in the routine clinical analysis of [123I]beta-CIT SPECT in patients with uncertain parkinsonism. PMID- 15817520 TI - Adult chronic sleepwalking and its treatment based on polysomnography. AB - Adult sleepwalking affects 2.5% of the general population and may lead to serious injuries. Fifty young adults with chronic sleepwalking were studied prospectively. Clinical evaluation, questionnaires from patients and bed partners, and polysomnography were obtained on all subjects in comparison with 50 age-matched controls. Subjects were examined for the presence of psychiatric anxiety, depression and any other associated sleep disorder. Isolated sleepwalking or sleepwalking with psychiatric disorders was treated with medication. All other patients with other sleep disorders were treated only for their associated problem. Prospective follow-up lasted 12 months after establishment of the most appropriate treatment. Patients with only sleepwalking, treated with benzodiazepines, dropped out of follow-up testing and reported persistence of sleepwalking, as did patients with psychiatric-related treatment. Chronic sleepwalkers frequently presented with sleep-disordered breathing (SDB). All these patients were treated only for their SDB, using nasal continuous positive airway pressure (CPAP). All nasal CPAP-compliant patients had control of sleepwalking at all stages of follow-up. Non-compliant nasal CPAP patients had persistence of sleepwalking. They were offered surgical treatment for SDB. Those successfully treated with surgery also had complete resolution of sleepwalking. Successful treatment of SDB, which is frequently associated with chronic sleepwalking, controlled the syndrome in young adults. PMID- 15817521 TI - Acute treatment with the PPARgamma agonist pioglitazone and ibuprofen reduces glial inflammation and Abeta1-42 levels in APPV717I transgenic mice. AB - Neuritic plaques in the brain of Alzheimer's disease patients are characterized by beta-amyloid deposits associated with a glia-mediated inflammatory response. Non-steroidal anti-inflammatory drug (NSAID) therapy reduces Alzheimer's disease risk and ameliorates microglial reactivity in Alzheimer's disease brains; however, the molecular mechanisms subserving this effect are not yet clear. Since several NSAIDs bind to and activate the nuclear receptor peroxisome proliferator activated receptor-gamma (PPARgamma) which acts to inhibit the expression of proinflammatory genes, this receptor appears a good candidate to mediate the observed anti-inflammatory effects. Recent data in vitro suggested that NSAIDs negatively regulate microglial activation and immunostimulated amyloid precursor protein processing via PPARgamma activation. We report that an acute 7 day oral treatment of 10-month-old APPV717I mice with the PPARgamma agonist pioglitazone or the NSAID ibuprofen resulted in a reduction in the number of activated microglia and reactive astrocytes in the hippocampus and cortex. Drug treatment reduced the expression of the proinflammatory enzymes cyclooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS). In parallel to the suppression of inflammatory markers, pioglitazone and ibuprofen treatment decreased beta secretase-1 (BACE1) mRNA and protein levels. Importantly, we observed a significant reduction of the total area and staining intensity of Abeta1-42 positive amyloid deposits in the hippocampus and cortex. Additionally, animals treated with pioglitazone revealed a 27% reduction in the levels of soluble Abeta1-42 peptide. These findings demonstrate that anti-inflammatory drugs can act rapidly to inhibit inflammatory responses in the brain and negatively modulate amyloidogenesis. PMID- 15817522 TI - Biological basis for human capacitation. AB - More than 50 years ago Austin and Chang defined mammalian sperm capacitation as a period of time that sperm must reside in the female reproductive tract before they acquire the ability to fertilize oocytes. Since then numerous investigations have attempted to more clearly define the molecules and processes that are a part of capacitation. The data that have provided a more clear definition of capacitation were primarily derived from in vitro experiments. This is particularly true for studies on human sperm capacitation. While ethical constraints have limited an equal balance of in vivo studies there are those data that when coupled with some of the in vitro data allow for the formulation of a biological framework for human sperm capacitation in vivo. This review will put forth the biological basis for human capacitation. PMID- 15817523 TI - Possible applications of a non-contact 1.48 microm wavelength diode laser in assisted reproduction technologies. AB - Recently, one laser system has been introduced in IVF fulfilling all safety requirements, while achieving a high standard of reproducibility in terms of ablation diameter. This 1.48 microm wavelength indium-gallium-arsenic-phosphorus (InGaAsP) semiconductor laser offers a variety of laser applications to the embryologist. On the one hand, zona pellucida of oocytes or embryos can be manipulated in order to facilitate ICSI or biopsy and assist hatching, and on the other, spermatozoa may be paralysed or immobilized prior to usage. To conclude, the 1.48 microm diode laser provides a promising tool for the microdissection of subcellular targets. The diode laser stands out due to the rapidity, the simplicity and the safety of the procedure which is supported by healthy offspring after laser application. PMID- 15817524 TI - Sex hormones and the immune response in humans. AB - In addition to their effects on sexual differentiation and reproduction, sex hormones appear to influence the immune system. This results in a sexual dimorphism in the immune response in humans: for instance, females produce more vigorous cellular and more vigorous humoral immune reactions, are more resistant to certain infections, and suffer a higher incidence of autoimmune diseases. Disease expression is also affected by the reproductive status of the female. As sex steroids--estrogens, progesterone and testosterone--differ between gender and within different reproductive stages, a lot of research has focussed on the effects of sex hormones on immune responses. Although there is also a vast literature on the effects of sex hormones on immune responses in animals, in this review we will focus on the most intriguing effects and mechanisms by which sex hormones affect different components of the immune system in humans. PMID- 15817525 TI - Efforts to create an artificial testis: culture systems of male germ cells under biochemical conditions resembling the seminiferous tubular biochemical environment. AB - Induction of meiotic and post-meiotic alterations of male germ cells in vitro has been the target of several research efforts since 1960. However, to date, the establishment of an ideal culture system in which spermatogonial stem cells can be maintained and directed to proliferate and undergo meiosis and complete spermiogenesis does not exist. This is attributed to the difficulties concerning the isolation and purification of defined subpopulations of germ cells and the establishment of male germ cell lines. In addition, there is no adequate knowledge regarding the optimal biochemical conditions that promote the survival and differentiation of germ cells in long-term cultures. This review focuses on the methodologies that have been proved sufficient to achieve differentiation of cultured male germ cells. Furthermore, the factors regulating spermatogenesis and the technical prerequisites to achieve differentiation of cultured male germ cells are described. Finally, the role of in vitro cultures of immature diploid germ cells in the therapeutic management of men negative for haploid cells in their testes and the subsequent potential genetic and epigenetic risks are discussed. PMID- 15817526 TI - Assessment of methodological quality of primary studies by systematic reviews: results of the metaquality cross sectional study. AB - OBJECTIVES: To describe how the methodological quality of primary studies is assessed in systematic reviews and whether the quality assessment is taken into account in the interpretation of results. DATA SOURCES: Cochrane systematic reviews and systematic reviews in paper based journals. STUDY SELECTION: 965 systematic reviews (809 Cochrane reviews and 156 paper based reviews) published between 1995 and 2002. DATA SYNTHESIS: The methodological quality of primary studies was assessed in 854 of the 965 systematic reviews (88.5%). This occurred more often in Cochrane reviews than in paper based reviews (93.9% v 60.3%, P < 0.0001). Overall, only 496 (51.4%) used the quality assessment in the analysis and interpretation of the results or in their discussion, with no significant differences between Cochrane reviews and paper based reviews (52% v 49%, P = 0.58). The tools and methods used for quality assessment varied widely. CONCLUSIONS: Cochrane reviews fared better than systematic reviews published in paper based journals in terms of assessment of methodological quality of primary studies, although they both largely failed to take it into account in the interpretation of results. Methods for assessment of methodological quality by systematic reviews are still in their infancy and there is substantial room for improvement. PMID- 15817527 TI - Comparison of descriptions of allocation concealment in trial protocols and the published reports: cohort study. AB - OBJECTIVES: To compare how allocation concealment is described in publications of randomised clinical trials and corresponding protocols, and to estimate how often trial publications with unclear allocation concealment have adequate concealment according to the protocol. DESIGN: Cohort study of 102 sets of trial protocols and corresponding publications. SETTING: Protocols of randomised trials approved by the scientific and ethical committees for Copenhagen and Frederiksberg, 1994 and 1995. MAIN OUTCOME MEASURES: Frequency of adequate, unclear, and inadequate allocation concealment and sequence generation in trial publications compared with protocols, and the proportion of protocols where methods were reported to be adequate but descriptions were unclear in the trial publications. RESULTS: 96 of the 102 trials had unclear allocation concealment according to the trial publication. According to the protocols, 15 of these 96 trials had adequate allocation concealment (16%, 95% confidence interval 9% to 24%), 80 had unclear concealment (83%, 74% to 90%), and one had inadequate concealment. When retrospectively defined loose criteria for concealment were applied, 83 of the 102 trial publications had unclear concealment. According to their protocol, 33 of these 83 trials had adequate allocation concealment (40%, 29% to 51%), 49 had unclear concealment (59%, 48% to 70%), and one had inadequate concealment. CONCLUSIONS: Most randomised clinical trials have unclear allocation concealment on the basis of the trial publication alone. Most of these trials also have unclear allocation concealment according to their protocol. PMID- 15817528 TI - "Right to die". PMID- 15817529 TI - Managing depression in primary care. PMID- 15817530 TI - Aortic stenosis. PMID- 15817531 TI - Long term sickness absence. PMID- 15817532 TI - New professional roles in surgery. PMID- 15817534 TI - NHS criticised for lax control over drugs industry. PMID- 15817535 TI - Government pledges 2.5m pounds sterling for academic medicine. PMID- 15817536 TI - 23% of babies in England are delivered by caesarean section. PMID- 15817538 TI - Assisted dying legislation must wait until after UK election. PMID- 15817539 TI - US scheme prevents some hospital admissions. PMID- 15817540 TI - Commission rejects mass genetic screening at birth. PMID- 15817543 TI - Sex inequalities in health care need tackling. PMID- 15817542 TI - International community has failed Sudan. PMID- 15817544 TI - Global ecological disaster predicted in next 50 years. PMID- 15817547 TI - Surgeons' ties to device manufacturers under investigation. PMID- 15817549 TI - Taking a collaborative approach to research. PMID- 15817550 TI - Prophylaxis and follow-up after possible exposure to HIV, hepatitis B virus, and hepatitis C virus outside hospital: evaluation of policy 2000-3. AB - PROBLEM: Prophylactic treatment and follow-up after exposure to HIV, hepatitis B, and hepatitis C outside hospital needs to be improved. BACKGROUND AND SETTING: Until January 2000, people in Amsterdam could report exposure outside hospital to either a hospital or the municipal health service. If they reported to the municipal health service, they were then referred to hospitals for HIV prophylaxis, whereas the municipal health service handled treatment and follow-up related to hepatitis B and hepatitis C and traced sources. For cases reported to a hospital, hospital staff often did not trace HIV sources or follow up patients for hepatitis B and hepatitis C. KEY MEASURES FOR IMPROVEMENT: Providing adequate treatment for HIV, hepatitis B and hepatitis C after exposure for all reported exposures outside hospital. STRATEGIES FOR CHANGE: On 1 January 2000, a new protocol was introduced in which three Amsterdam hospitals and the municipal health service collaborated in the treatment and follow-up of exposures outside hospital. Both municipal health service and hospitals can decide whether HIV prophylaxis is necessary and prescribe accordingly. All people exposed in the community who report to hospitals are subsequently referred to the municipal health service for further treatment and follow-up. EFFECTS OF CHANGE: The protocol is effective in that most people comply with treatment and follow-up. When indicated, HIV prophylaxis is started soon after exposure. In nearly two thirds of cases the municipal health service traced and tested the source. LESSONS LEARNT: Provision of treatment and follow-up in one place enables treatment, tracing and testing sources, and follow-up, including counselling and registration of all reported exposures in Amsterdam, which allows for swift identification of emerging epidemiological trends. Since May 2004 all Amsterdam hospitals have participated in the protocol. PMID- 15817551 TI - Necrotising fasciitis. PMID- 15817552 TI - Postoperative hypoxia in a woman with Down's syndrome: case presentation. PMID- 15817553 TI - Common mental health problems. PMID- 15817554 TI - Managing common mental health disorders in primary care: conceptual models and evidence base. PMID- 15817555 TI - Treatment allocation by minimisation. PMID- 15817556 TI - Non-specific effects of vaccination: survival bias may explain findings. PMID- 15817557 TI - Non-specific effects of vaccination: vaccines have non-specific (heterologous) effects. PMID- 15817558 TI - DTP in low income countries: improved child survival or survival bias? PMID- 15817559 TI - Use of chaperones in general practice: chaperones protect both parties. PMID- 15817560 TI - Use of chaperones in general practice: GPs try to balance doctors' and patients' needs. PMID- 15817561 TI - Phenobarbital for epilepsy: much is still to be learnt. PMID- 15817562 TI - Ethics and research governance in a multicentre study: add 150 days to your study protocol. PMID- 15817564 TI - Species-independent detection of RNA virus by representational difference analysis using non-ribosomal hexanucleotides for reverse transcription. AB - A method for the isolation of genomic fragments of RNA virus based on cDNA representational difference analysis (cDNA RDA) was developed. cDNA RDA has been applied for the subtraction of poly(A)(+) RNAs but not for poly(A)(-) RNAs, such as RNA virus genomes, owing to the vast quantity of ribosomal RNAs. We constructed primers for inefficient reverse transcription of ribosomal sequences based on the distribution analysis of hexanucleotide patterns in ribosomal RNA. The analysis revealed that distributions of hexanucleotide patterns in ribosomal RNA and virus genome were different. We constructed 96 hexanucleotides (non ribosomal hexanucleotides) and used them as mixed primers for reverse transcription of cDNA RDA. A synchronous analysis of hexanucleotide patterns in known viral sequences showed that all the known genomic-size viral sequences include non-ribosomal hexanucleotides. In a model experiment, when non-ribosomal hexanucleotides were used as primers, in vitro transcribed plasmid RNA was efficiently reverse transcribed when compared with ribosomal RNA of rat cells. Using non-ribosomal primers, the cDNA fragments of severe acute respiratory syndrome coronavirus and bovine parainfluenza virus 3 were efficiently amplified by subtracting the cDNA amplicons derived from uninfected cells from those that were derived from virus-infected cells. The results suggest that cDNA RDA with non-ribosomal primers can be used for species-independent detection of viruses, including new viruses. PMID- 15817563 TI - Proximity extension of circular DNA aptamers with real-time protein detection. AB - Multivalent circular aptamers or 'captamers' have recently been introduced through the merger of aptameric recognition functions with the basic principles of DNA nanotechnology. Aptamers have strong utility as protein-binding motifs for diagnostic applications, where their ease of discovery, thermal stability and low cost make them ideal components for incorporation into targeted protein assays. Here we report upon a property specific to circular DNA aptamers: their intrinsic compatibility with a highly sensitive protein detection method termed the 'proximity extension' assay. The circular DNA architecture facilitates the integration of multiple functional elements into a single molecule: aptameric target recognition, nucleic acid hybridization specificity and rolling circle amplification. Successful exploitation of these properties is demonstrated for the molecular analysis of thrombin, with the assay delivering a detection limit nearly three orders of magnitude below the dissociation constants of the two contributing aptamer-thrombin interactions. Real-time signal amplification and detection under isothermal conditions points towards potential clinical applications, with both fluorescent and bioelectronic methods of detection achieved. This application elaborates the pleiotropic properties of circular DNA aptamers beyond the stability, potency and multitargeting characteristics described earlier. PMID- 15817565 TI - Substrate discrimination in RNase P RNA-mediated cleavage: importance of the structural environment of the RNase P cleavage site. AB - Like the translational elongation factor EF-Tu, RNase P interacts with a large number of substrates where RNase P with its RNA subunit generates tRNAs with matured 5' termini by cleaving tRNA precursors immediately 5' of the residue at +1, i.e. at the position that corresponds to the first residue in tRNA. Most tRNAs carry a G+1C+72 base pair at the end of the aminoacyl acceptor-stem whereas in tRNA(Gln) G+1C+72 is replaced with U+1A+72. Here, we investigated RNase P RNA mediated cleavage as a function of having G+1C+72 versus U+1A+72 in various substrate backgrounds, two full-size tRNA precursors (pre-tRNA(Gln) and pre tRNA(Tyr)Su3) and a model RNA hairpin substrate (pATSer). Our data showed that replacement of G+1C+72 with U+1A+72 influenced ground state binding, cleavage efficiency under multiple and single turnover conditions in a substrate-dependent manner. Interestingly, we observed differences both in ground state binding and rate of cleavage comparing two full-size tRNA precursors, pre-tRNA(Gln) and pre tRNA(Tyr)Su3. These findings provide evidence for substrate discrimination in RNase P RNA-mediated cleavage both at the level of binding, as previously observed for EF-Tu, as well as at the catalytic step. In our experiments where we used model substrate derivatives further indicated the importance of the +1/+72 base pair in substrate discrimination by RNase P RNA. Finally, we provide evidence that the structural architecture influences Mg2+ binding, most likely in its vicinity. PMID- 15817566 TI - Vertebrate telomere repeat DNAs favor external loop propeller quadruplex structures in the presence of high concentrations of potassium. AB - The circular dichroism, CD, spectra of the telomere repeats of vertebrates, d(TTAGGG), indicate that parallel type quadruplex structures or disordered single stranded structures are formed in low salt. Anti-parallel quadruplex structures are favored in the presence of high concentrations, 140 mM, of sodium. External loop, also known as propeller, parallel type structures are favored in the presence of high concentrations, 100 mM, of potassium in the presence of either 5 or 140 mM sodium. The cation dependence of the CD spectra of the vertebrate telomere repeat DNAs is distinctly different from that of the telomere repeats of Tetrahymena and Oxytricha as well as that of the thrombin binding aptamer. These results indicate that the external loop structures may be present in vertebrate telomeres under the conditions of high potassium and low sodium concentration found in nuclei. PMID- 15817567 TI - Retropseudogenes derived from the human Ro/SS-A autoantigen-associated hY RNAs. AB - We report the characterization in the human genome of 966 pseudogenes derived from the four human Y (hY) RNAs, components of the Ro/SS-A autoantigen. About 95% of the Y RNA pseudogenes are found in corresponding locations on the chimpanzee and human chromosomes. On the contrary, Y pseudogenes in mice are both infrequent and found in different genomic regions. In addition to this rodent/primate discrepancy, the conservation of hY pseudogenes relative to hY genes suggests that they occurred after rodent/primate divergence. Flanking regions of hY pseudogenes contain convincing evidence for involvement of the L1 retrotransposition machinery. Although Alu elements are found in close proximity to most hY pseudogenes, these are not chimeric retrogenes. Point mutations in hY RNA transcripts specifically affecting binding of Ro60 protein likely contributed to their selection for direct trans retrotransposition. This represents a novel requirement for the selection of specific RNAs for their genomic integration by the L1 retrotransposition machinery. Over 40% of the hY pseudogenes are found in intronic regions of protein-coding genes. Considering the functions of proteins known to bind subsets of hY RNAs, hY pseudogenes constitute a new class of L1 dependent non-autonomous retroelements, potentially involved in post transcriptional regulation of gene expression. PMID- 15817568 TI - Site-specific reverse splicing of a HEG-containing group I intron in ribosomal RNA. AB - The wide, but scattered distribution of group I introns in nature is a result of two processes; the vertical inheritance of introns with or without losses, and the occasional transfer of introns across species barriers. Reversal of the group I intron self-splicing reaction, termed reverse splicing, coupled with reverse transcription and genomic integration potentially mediate an RNA-based intron mobility pathway. Compared to the well characterized endonuclease-mediated intron homing, reverse splicing is less specific and represents a likely explanation for many intron transpositions into new genomic sites. However, the frequency and general role of an RNA-based mobility pathway in the spread of natural group I introns is still unclear. We have used the twin-ribozyme intron (Dir.S956-1) from the myxomycete Didymium iridis to test how a mobile group I intron containing a homing endonuclease gene (HEG) selects between potential insertion sites in the small subunit (SSU) rRNA in vitro, in Escherichia coli and in yeast. Surprisingly, the results show a site-specific RNA-based targeting of Dir.S956-1 into its natural (S956) SSU rRNA site. Our results suggest that reverse splicing, in addition to the established endonuclease-mediated homing mechanism, potentially accounts for group I intron spread into the homologous sites of different strains and species. PMID- 15817569 TI - Argonaute protein PIWI controls mobilization of retrotransposons in the Drosophila male germline. AB - Proteins of the Argonaute family have been identified as key components of RNA interference (RNAi) pathway. RNAi-related mechanisms are implicated in the regulation of gene expression and repression of transposable elements in eukaryotes. The piwi gene encoding protein of the Drosophila Argonaute family was shown to be required for the germ stem cells maintenance. Here, we show that piwi is involved in silencing of LTR retrotransposons in testes. piwi mutations led to derepression of endogenous retrotransposon copia as well as to upregulation of the reporter gene driven by copia LTR. piwi mutation causes accumulation of retrotransposon mdg1 transcripts at the apical tip of testes, including germinal proliferative center where PIWI protein was shown to be expressed. We applied inverse PCR approach to detect the newly arisen insertions of the mdg1 retrotransposon in the progeny of individual piwi mutant males. Owing to piwi mutation a high rate of mdg1 transpositions was revealed. Thus, piwi is involved in the silencing of retrotransposons in the precursors of male gametes. Our results provide the first evidence that protein of the Argonaute family prevents retrotranspositions. It is supposed that the disturbance of RNA silencing system in germinal cells might cause transposition burst. PMID- 15817571 TI - Comparative genotoxic evaluation of 2-furylethylenes and 5-nitrofurans by using the comet assay in TK6 cells. AB - The genotoxicity of three 2-furylethylene derivatives and four 5-nitrofurans was evaluated by using the comet assay in human lymphoblastoid cultured TK6 cells. The 2-furylethylene derivatives were 2-furyl-1-nitroethene, 1-(5-bromofur-2-yl)-2 nitroethene and 1-(5-bromofur-2-yl)-2-bromo-2-nitroethene, while the 5 nitrofurans were nitrofurantoin, nitrofurazone, furazolidone and 5-nitro-2 furanacrolein. The treatments lasted for 3 h in the absence of metabolic activation. No genotoxic effects were observed for two of the 2-furylethylene compounds, while the derivative 1-(5-bromofur-2-yl)-2-nitroethene showed a statistically significant response mainly at the highest concentration tested; this effect was considered biologically relevant and the compound was classified as slightly genotoxic. On the other hand, for the classical 5-nitrofurans tested there is a tendency towards a dose-related increase of the DNA damage in the comet assay and the observed increases for the parameters analysed (Olive tail moment, tail % DNA and tail length) were significant for all compounds. Then, the four 5-nitrofurans tested were considered genotoxic. These results show that the position of the nitro group influences the genotoxicity of the assayed compounds. Thus, in this comet assay, the 2-furylethylene derivatives having the nitro group attached outside the furan ring appear to be much less genotoxic than the 5 nitrofurans. PMID- 15817572 TI - Influence of caspase activity on micronuclei detection: a possible role for caspase-3 in micronucleation. AB - Aneugenic and clastogenic agents are good inducers of both micronuclei and apoptosis. In its turn, apoptosis may modify the threshold values for the induction of micronuclei. This is of major concern for accurate assessment of hazard related to exposure to mutagens. In the present work we studied the influence of caspases, the key regulators of the apoptotic process, on the induction of micronuclei in the cytokinesis block micronucleus assay. For this, we applied a combined approach in which both human peripheral blood mononucleated cells (PBMC) and the paired human breast carcinoma cell lines MCF-7, which is caspase-3 deficient, and the caspase-3 transfected MCF-7 (MCF-7casp-3) were used to study the influence of caspase activity on micronuclei. When nocodazole induced apoptosis was inhibited by the use of inhibitors of the two main apical caspases-8 and -9 in PBMC, the frequencies of micronucleated binucleates (MNCB) increased with inhibition of these caspases confirming that apoptosis can eliminate micronucleated cells. On the contrary when caspase-3 was inhibited, the frequencies of MNCB was lower, suggesting a role of caspase-3, also in micronuclei formation. To verify this hypothesis, we compared the induction of apoptosis and micronuclei by the aneugen nocodazole, the clastogen methyl methane sulfonate (MMS) and the non-mutagenic apoptogen staurosporin in MCF-7 and MCF 7casp-3 cells. The results showed that when caspase-3 activity was impaired, in the parental MCF-7 cell line or in the MCF-7casp-3 cells in the presence of the caspase-3 inhibitor, the frequencies of nocodazole or MMS induced micronuclei decreased. These results suggest that caspase-3, besides its function as an effector caspase in the apoptotic pathway, is also involved in the formation of micronuclei. PMID- 15817573 TI - Incidence of cytogenetic damage in lead-zinc mine workers exposed to radon. AB - The purpose of this study was to detect cytogenetic damage in mine workers working in a lead-zinc mine, which could be associated with a combined exposure to radon and heavy metals. Our study involved 70 mine workers from the lead-zinc mine. We used peripheral blood lymphocytes as the target material. The total share of structural chromosome aberration (SCA) decreased significantly over the 3 years of monitoring, from 5.08/200 analyses of metaphases in 1995 to 3.28 in 1997, owing to the decrease in exposure during the process of mine closure. The share of SCA was significantly different from the group of local people, who had never worked in the mine (1.43), as well as from the control group of Slovene residents (1.88). The share of micronuclei (MN) in mine workers also decreased in the monitored period, from 14.65/500 cytokinesis-blocked cells in 1995 to 11.77 in 1997, while the sister chromatic exchange (SCE) level did not change much (from 8.105/50 analysed cells in 1995 to 7.73 in 1997). Owing to the closure activities, the received concentrations of contaminants were falling constantly, particularly concentrations of radon. This was particularly evident in the level of SCA and the MN incidence, while the SCE values remained nearly on the same level. This indicates that the incidence of SCE is probably more strongly influenced by heavy metals than by radon. PMID- 15817574 TI - DNA damage induced by a quinoxaline 1,4-di-N-oxide derivative (hypoxic selective agent) in Caco-2 cells evaluated by the comet assay. AB - The DNA damage induced by 7-chloro-3-[[(N,N-dimethylamino)propyl]amino]-2 quinoxalinecarbonitrile 1,4-di-N-oxide hydrochloride (Q-85 HCl) in Caco-2 cells under hypoxic and well-oxygenated conditions has been studied by using the comet assay. This compound has shown a good in vitro profile of high selective toxicity in hypoxia, but its mechanism of action is unknown. The DNA damage has been evaluated by performing the comet assay after a 2-h treatment with Q-85 HCl (0.1, 0.2, 0.4 microM in hypoxia; 20, 40 microM in well-oxygenated conditions). The number of cells in apoptosis has also been assessed by flow cytometry analysis of Annexin V-FITC staining. The capability of the cells to repair the DNA damage and the proliferation rate was evaluated at different times after the treatment (24 168 h). Under hypoxic conditions, a clear dose-dependent increase in the number of nuclei with a comet was observed (comet score: 132 +/- 13, 343 +/- 30 and 399 +/- 1; control comet score: 42 +/- 14). Under well-oxygenated conditions, the number of nuclei with comet increased significantly with respect to the control (comet score: 273 +/- 14 and 312 +/- 9; control comet score: 27 +/- 4). Cells in apoptosis were not detected by the comet assay nor by flow cytometry. The recovery from DNA damage was time- and concentration-dependent in hypoxia (cells treated with the highest concentration still showed DNA damage after 72 h) and rather time-dependent in well-oxygenated conditions (DNA was completely repaired after 24 h). In conclusion, Q-85 HCl acts by DNA damage and not only the reduced intermediate is genotoxic but also some other derivatives and Q-85 HCl itself may be acting. PMID- 15817575 TI - Interaction between cadmium and aromatic DNA adducts in hprt mutagenesis during foetal development. AB - The foetus is exposed to multiple xenobiotics through the mother's circulation and this is possibly involved in the development of diseases in later life. Heavy metals and lipophilic genotoxins in umbilical cord blood of newborns may have synergistic effects on mutagenesis in the hypoxanthine-phosphoribosyltransferase (HPRT) reporter gene. Concentrations of zinc (Zn), lead (Pb) and cadmium (Cd) were determined in the peripheral and cord blood of 16 non-smoking and 9 smoking healthy mothers by atomic absorption spectrometry. Lipophilic DNA adducts in lymphocytes were determined in the same subjects by 32P-postlabelling and the HPRT-variant frequency was assessed by the evaluation of 6-thioguanine resistant cells. Although the Cd/Zn ratio was 2.5-fold higher in the blood of smoking women than in non-smoking women (1.0 +/- 0.2 and 0.4 +/- 0.1, respectively, P = 0.007), this difference could not be observed in umbilical cord blood (0.3 +/- 0.1 and 0.3 +/- 0.1, respectively, P = 0.66). Similarly, mean DNA adduct levels were increased in the lymphocytes of smoking women compared with non-smoking controls (0.99 +/- 0.31 adducts/10(8) nt and 0.43 +/- 0.12, respectively, P = 0.009), but were only marginally higher in the neonates of smokers than in their non-smoking counterparts (0.57 +/- 0.29 and 0.24 +/- 0.09, respectively, P = 0.38). Since Cd is known to effectively inhibit DNA repair, we hypothesized that concomitant exposure of neonates to Cd and genotoxic compounds may result in an increased fixation of DNA damage into somatic mutations. Indeed, the number of HPRT variants per adduct (i.e. the mutagenic efficiency of adducts) correlated positively with the Cd concentrations in cord blood (r = 0.61, P = 0.001). These data suggest a molecular link between DNA damage, inhibition of DNA repair by Cd and in vivo mutagenesis during foetal development. Thus, exposure to heavy metals may enhance the mutagenic potential of DNA-damaging compounds and results in biologically relevant genotoxic effects in neonates. PMID- 15817576 TI - Measurements of finger doses in x-ray guided surgery, nuclear medicine and research. AB - The Norwegian Radiation Protection Authority has performed measurements of finger doses to nuclear medicine staff exposed to 99Tc(m), researchers handling 32P, surgeons performing X-ray guided orthopaedic surgery and surgeons and radiologists performing X-ray guided endovascular treatment of abdominal aortic aneurysms (AAA). Calibrations were done with X-ray qualities N-40, N-60 and N-300 and with the beta source 90Sr + 90Y. Annual doses were estimated for the nuclear medicine staff and the orthopaedic surgeons. The mean annual finger dose to nuclear medicine staff exposed to 99Tc(m) was estimated to be 18.8 mSv, and the mean annual finger dose to surgeons performing X-ray guided orthopaedic surgery was 13.7 mSv. The surgeons and radiologists performing X-ray guided endovascular treatment of AAA received a mean finger dose of 0.35 mSv per treatment. The majority of researchers handling 32P received no finger dose at all, and the maximum reading was 1.65 mSv. All occupational groups received finger doses well below the annual finger dose limit of 500 mSv. PMID- 15817577 TI - Comparative study of Dutch and German emergency-management models for near border nuclear accidents. AB - Four institutes, all of which are involved in nuclear-emergency management in the Dutch-German border region, have compared their short-range dispersion and radiological dose models using scenarios consisting of single-station meteorology and two dispersed radionuclides. After adjustment of some of the parameters, the consequence of the differences in parameters on the effective dose was quantified at several stages from source to exposure. Results for the neutral stability class agreed within a factor of four. Variations in wet deposition of radioactive material, giving rise to external radiation from the ground, can cause significant variations to the effective dose. Furthermore, the way the different emergency-management tools model the atmospheric dispersion for a stable stability class in the horizontal plane can generate large differences. Finally, the methodology of calculating cloudshine is not comparable among the models, which causes the effective dose near the source to show large deviations for high emission sources. PMID- 15817578 TI - Evaluation of etching correction factor for LR115 cellulose nitrate films from track parameters. AB - LR115 cellulose nitrate films efficiency is strongly dependent on the conditions of etching. In this paper a new method to evaluate a correction factor for this effect is given. The film is scanned with an high-resolution scanner, the diameter of the tracks is measured, and the correction factor is calculated from the average diameter of the tracks. The sensitivity of the film is found to have a linear dependence on the average diameter, and the classical correction, based on the residual thickness, can be substituted by a method based on average track diameter. PMID- 15817579 TI - Measurements of activity concentrations of naturally occurring radionuclides in soil samples from Punjab province of Pakistan and assessment of radiological hazards. AB - The natural level of radioactivity in soil is one of the major causes of external gamma rays exposure. It was considered imperative to measure gamma-ray activity concentrations due to naturally occurring, potentially hazardous radionuclides from 232Th, 226Ra and 40K in soil samples from Punjab, the most populated province of Pakistan. The activity concentrations were measured in soil samples collected from all 34 district headquarters of Punjab province using a P-type coaxial high purity germanium (HPGe)-based gamma-ray spectrometer having a relative efficiency of 60%. The mean activity concentrations for 232Th, 226Ra and 40K were found to be 41 +/- 8, 35 +/- 7 and 615 +/- 143 Bq kg(-1), respectively in soil samples from all 34 district headquarters of Punjab. The mean values of radium equivalent index (Ra(eq)) and total annual external gamma radiation dose were found to be 141 +/- 27 Bq kg(-1)and 0.38 mSv, respectively. Since these estimated radiological impact assessment factors are lower than the recommended values there seems to be no potential radiological health hazard associated with the soils from the district headquarters in the Punjab province of Pakistan. Geological studies show that most of the soils in Punjab have developed on the extensive fluviatile deposits of five rivers and a stable ratio of natural activity concentrations is observed in soil samples of the areas under study. PMID- 15817580 TI - Effects of subfertility cause, smoking and body weight on the success rate of IVF. AB - BACKGROUND: We investigated the separate and combined effects of smoking and body mass index (BMI) on the success rate of IVF for couples with different causes of subfertility. METHODS: The success rate of IVF was examined in 8457 women. Detailed information on reproduction and lifestyle factors was combined with medical record data on IVF treatment. All IVF clinics in The Netherlands participated in this study. The main outcome measures were live birth rate per first cycle of IVF differentiated for the major predictive factors. RESULTS: For male subfertility the delivery rate per cycle was significantly lower than unexplained subfertility, OR of 0.70 (95% CI 0.57-0.86); for tubal pathology, the delivery rate was slightly lower, OR = 0.86 (95% CI 0.70-1.01). Smoking was associated with a significantly lower delivery rate was slightly lower; for OR = 0.72 (95% CI 0.61-0.84) and a significantly higher abortion rate compared to non smoking delivery rates of 21.4% and 16.4%, respectively (P=0.02). Women with a BMI of > or = 27 kg/m2 had a significantly lower delivery rate, with an OR of 0.67 (95% CI 0.48-0.94), compared with normal weight women (BMI > or = 20 and <27 kg/m2). CONCLUSIONS: Both smoking and overweight unfavourably affect the live birth rate after IVF. The devastating impact of smoking on the live birth rate in IVF treatment is comparable with an increase in female age of >10 years from age 20 to 30 years. Subfertile couples may improve the outcome of IVF treatment by lifestyle changes. PMID- 15817581 TI - Acute renal failure following IVF: case report. AB - IVF is one of the most comprehensively registered interventions in clinical medicine. IVF is regarded as safe with very few complications. We report a woman who developed acute renal failure due to compression of both ureters from enlarged stimulated ovaries. The condition was diagnosed using ultrasound and magnetic resonance imaging (MRI). The condition was treated with insertion of double-J stents in both ureters and resolved without need of dialysis. Compression of the ureters due to enlarged ovaries should be considered if a patient develops acute renal failure following IVF. PMID- 15817582 TI - Low-dose aspirin does not improve ovarian responsiveness or pregnancy rate in IVF and ICSI patients: a randomized, placebo-controlled double-blind study. AB - BACKGROUND: Poor ovarian and endometrial responses to gonadotrophin stimulation in assisted reproduction techniques lead to decreased pregnancy rates. The aim of the present study was to test the hypothesis that low-dose aspirin started prior to controlled ovarian stimulation improves ovarian responsiveness, pregnancy rate (PR) and pregnancy outcome. METHODS: A total of 374 women who were to undergo IVF/ICSI were randomized to receive 100 mg of aspirin (n=186) or placebo (n=188) daily. Treatment was started on the first day of controlled ovarian stimulation. It was continued until menstruation or a negative pregnancy test. Pregnant women continued the medication until delivery. The main outcome measures were the number of oocytes, number and quality of embryos, the clinical PR and pregnancy outcome. RESULTS: The mean (+/-SD) number of oocytes (12.0+/-7.0 versus 12.7+/ 7.2), the total mean number of embryos (5.82+/-4.35 versus 5.99+/-4.66), the mean number of top quality embryos (0.99+/-1.39 versus 1.18+/-1.51) and the number of embryos transferred (1.64+/-0.64 versus 1.63+/-0.71) did not differ in the aspirin and placebo groups. Between the aspirin and placebo group, there was no statistically significant difference in clinical PR per embryo transfer (25.3%, n=44 out of 174 versus 27.4%, n=48 out of 175) or clinical PR per cycle initiated (23.7% versus 25.5%). Birth rate per embryo transfer did not differ significantly between the aspirin (18.4%) and placebo (21.1%) groups. The incidence of poor responders [12 (6.5%) versus 13 (6.9%)] was similar in both groups. CONCLUSIONS: The present results indicate that low-dose aspirin treatment does not have any beneficial effect on ovarian responsiveness, PR and pregnancy outcome in unselected women undergoing IVF/ICSI. PMID- 15817583 TI - Fertility and ovarian function are preserved in women treated with an intensified regimen of cyclophosphamide, adriamycin, vincristine and prednisone (Mega-CHOP) for non-Hodgkin lymphoma. AB - BACKGROUND: Intensive chemotherapy is widely used to improve the outcome of aggressive non-Hodgkin lymphoma (NHL). Since these regimens may cause premature ovarian failure (POF), the ovarian function was studied in 13 consecutive women aged < or =40 years, treated with four cycles of intensified CHOP (cyclophosphamide 2000-3000 mg/m2 per cycle doxorubicin 50 mg/m2, vincristine 1.4 mg/m2 (maximum 2 mg) and prednisone 100 mg/day were given every 3 weeks). METHODS: Patients aged <60 years with aggressive NHL were eligible for participating in a non-randomized phase II study if they had stage I, II, B, bulky, or stages III, IV disease with the age-adjusted international prognostic index of low-intermediate to high-risk score. Seven patients were concomitantly treated with D-TRP6-GnRH analogue (Decapeptyl; Ferring, Germany) for minimizing gonadal toxicity. RESULTS: With a median follow-up of 70 months only one patient had POF, while 12 patients retained fertility and eight conceived spontaneously delivering 12 healthy babies. CONCLUSION: It appears that high-dose cyclophosphamide does not affect the ovarian function or fertility in patients exposed to this medication during four consecutive cycles of intensified CHOP. PMID- 15817584 TI - A longitudinal, prospective study on emotional adjustment before, during and after consecutive fertility treatment cycles. AB - BACKGROUND: A longitudinal study into the course of the emotional response to IVF from pre-treatment to 6 months post-treatment and factors that contributed to that course. METHODS: A total of 148 IVF patients and 71 partners completed self report questionnaires on anxiety, depression, personality characteristics, meaning of fertility problems, coping, marital relationship and social support at pre-treatment. Assessments of anxiety and depression were repeated immediately following the final treatment cycle and again 6 months later (follow-up). RESULTS: Women showed an increase of both anxiety and depression after unsuccessful treatment and a decrease after successful treatment. Men showed no change in anxiety and depression either after successful or after unsuccessful treatment. In the 6 months after unsuccessful treatment, women showed no recovery. At follow-up, >20% of the women showed subclinical forms of anxiety and/or depression. Personality characteristics, meaning of the fertility problems, and social support determined the course of the emotional response. CONCLUSIONS: Most women adjusted well to unsuccessful treatment, but at follow up, a considerable proportion still showed substantial emotional problems. Personality characteristics, pre-treatment meaning of the fertility problems and social support have demonstrated the adjustment to unsuccessful IVF in women. This allows early identification of women at risk as well as tailored interventions. PMID- 15817585 TI - Clinical adaptation of the sperm ubuquitin tag immunoassay (SUTI): relationship of sperm ubiquitylation with sperm quality in gradient-purified semen samples from 93 men from a general infertility clinic population. AB - BACKGROUND: The proteolytic chaperone peptide ubiquitin accumulates in defective human spermatozoa. Immunodetection of ubiquitin in human sperm samples correlates with semen quality and male fertility. METHODS: Semen samples from 93 men from couples seeking infertility treatment were separated on a PureSperm density gradient and screened by immunofluorescence microscopy with anti-ubiquitin antibodies. The percentage of spermatozoa with head ubiquitylation was recorded and compared with clinical semen evaluation and embryo development data after IVF or ICSI. Subjects were divided into the following four groups based on the initial clinical diagnosis of the couples; group 1, male factor; group 2, idiopathic infertility; group 3, female infertility with neither partner having children previously; and group 4, female infertility with male partners having children from previous relationships. RESULTS: The percentage of sperm with ubiquitylated heads remaining after PureSperm separation in the respective groups was 4.0% (male factor), 2.5% (idiopathic infertility), 0.7% (female infertility and presumed fertile male) and 0.9% (female infertility with established fertile male). Negative correlations between sperm ubiquitin and several parameters reflective of embryo development after assisted fertilization were found within the male factor group. CONCLUSIONS: Use of this simplified ubiquitin-based sperm quality assay is feasible in a clinical environment. Since the gradient separation does not completely deplete the defective spermatozoa, the modified light microscopic sperm ubiquitin tag immunoassay could add a new level of stringency to the selection of human spermatozoa for ICSI. PMID- 15817586 TI - The 2004 Italian legislation regulating assisted reproduction technology: a multicentre survey on the results of IVF cycles. AB - BACKGROUND: The new Italian law, passed in 2004, regulating assisted reproduction technology imposes that no more than three oocytes can be fertilized at one time and that all embryos obtained must be transferred simultaneously. Oocyte cryopreservation is allowed while embryo cryostorage is banned. The aim of this study was to evaluate the clinical impact of these limitations. METHODS: Seven Italian infertility centres were invited to collect data on IVF cycles performed over the first 4 months of application of the new legislation. As a control, all centres provided data on cycles performed in the same solar period, 1 year before. RESULTS: Data from 1861 cycles were obtained, 961 in the pre-law period and 900 in the post-law period. Pregnancy rate per oocyte retrieval and rate of multiple pregnancies in the pre- and post-law periods were 27.0 and 24.2% (P=0.18) and 25.8 and 20.9% (P=0.11) respectively. However, the prohibition to freeze embryos does appear to have markedly reduced the cumulative rate of success. CONCLUSIONS: The rate of success of IVF-ICSI cycles using fresh embryos is not significantly influenced by the new legislation while the prohibition to freeze embryos seems to result in a more relevant impact. PMID- 15817587 TI - Periovulatory increase in temperature difference within the rabbit oviduct. AB - BACKGROUND: Earlier studies demonstrated a small temperature difference between the sperm storage and fertilization sites within the oviducts of rabbits and pigs. Our aim was to reveal the time dependence of this temperature difference relative to ovulation, and to determine how this difference is generated-by temperature elevation at one of these sites or by temperature decrease at the other site. METHODS: The temperature at the sperm storage site (at the isthmus near the uterotubal junction) and at the fertilization site (the isthmic ampullary junction) of rabbit oviducts were measured before, during, and after ovulation by two probes, connected to digital thermometers. Rectal temperature was constantly measured and served as a control for body temperature. RESULTS: The temperature difference between the fertilization site and the storage site was 0.8+/-0.2 degrees C before ovulation. This difference increased at ovulation, reaching 1.6+/-0.1 degrees C after ovulation (P<0.03). This increased difference was mainly due to temperature decrease in the sperm storage site. CONCLUSION: The temperature-difference increase within the rabbit oviduct is generated at ovulation by a reduced temperature at the sperm storage site. This temperature gradient may play a role in mammalian reproduction via sperm thermotaxis. PMID- 15817588 TI - Developmental incompetency of denuded mouse oocytes undergoing maturation in vitro is ooplasmic in nature and is associated with aberrant Oct-4 expression. AB - BACKGROUND: Germinal vesicle (GV) oocytes constitute a potential resource but their developmental competence is questionable especially when surrounding cumulus cells are removed. The intercellular factors/mechanisms underlying such poor embryonic competence may originate at a nuclear and/or ooplasmic level. METHODS: Immature or mature oocytes were obtained from three mouse strains following pregnant mare serum gonadotropin (PMSG) or PMSG+ human chorionic gonadotropin (hCG) treatment. Immature oocytes were denuded of cumulus cells prior to in vitro maturation. Pronuclear (PN) transfer was used to examine nuclear-ooplasmic interplay on resultant embryonic development and Oct-4 immuno staining patterns. RESULTS: Embryos arising from ooplasts of in vivo matured oocytes displayed significant increases in blastocyst formation rates and total blastomere numbers when compared to those created from ooplasts of denuded oocytes. Oct-4 staining was more pronounced and restricted to the inner cell mass (ICM) in blastocysts arising from the ooplasm of in vivo matured zygotes than in those created from denuded oocytes. CONCLUSIONS: Developmental defect(s) appear to develop primarily in the ooplasm of oocytes that are denuded of their cumulus cells prior to in vitro maturation. Such oocytes result in embryos with poor developmental competence. These defects result in anomalies in cell number and Oct-4 expression during the morula-blastocyst developmental transition. PMID- 15817589 TI - Oxidative stress and endometriosis. AB - BACKGROUND: Little is known about the aetiology of endometriosis; however, in the presence of oxidative stress, reactive oxygen species might increase growth and adhesion of endometrial cells in the peritoneal cavity, leading to endometriosis and infertility. Within a study investigating persistent organic compounds and endometriosis, the authors evaluated the association between oxidative stress and endometriosis. METHODS: Women aged 18-40 years who were undergoing laparoscopy were contacted to participate in the study (n = 100); 84 were eligible and agreed to be interviewed; 78 provided blood specimens. Four markers of oxidative stress and antioxidant status were measured in serum for 61 women. Multiple imputation of missing data was used to generate values for the missing oxidative stress data. RESULTS: Thirty-two women had visually confirmed endometriosis at laparoscopy while 52 did not, including 22 undergoing tubal ligation and 30 with idiopathic infertility. There was a weak association between thiobarbituric acid reactive substances (nmol/ml) and endometriosis, after adjusting for age, body mass index, current smoking, hormone use in the past 12 months, gravidity, serum vitamin E, serum estradiol, and total serum lipids (beta = 1.18; 95% CI-0.04, 2.39). CONCLUSIONS: These results suggest that oxidative stress might play a role in the development and progression of endometriosis, which should be evaluated in larger studies. PMID- 15817590 TI - Oral and pulmonary delivery of FSH-Fc fusion proteins via neonatal Fc receptor mediated transcytosis. AB - BACKGROUND: The alpha and beta subunits of FSH were fused to the Fc domain of IgG1 either in a single chain or a heterodimer format. These molecules were absorbed through the epithelium in lung and intestine by neonatal Fc receptor (FcRn)-mediated transcytosis. METHODS AND RESULTS: Single chain and heterodimer FSH-Fc were made recombinantly in Chinese hamster ovary cells. Treatment of rats with a single s.c. dose of single chain or heterodimer FSH-Fc resulted in greater stimulation of ovarian weight (20.8+/-3.9 and 26.9+/-6.1 mg respectively) compared to those receiving vehicle (12.1+/-1.0 mg) or an equimolar dose of recombinant human FSH (14.3+/-1.7 mg). Both FSH-Fc fusion proteins were absorbed after oral dosing of newborn rats with long terminal half-lives of approximately 60 h, and pulmonary delivery in four cynomolgus monkeys produced maximum serum concentrations between 69 and 131 ng/ml with long terminal half-lives between 55 and 210 h. Serum inhibin levels increased after pulmonary dosing with single chain FSH-Fc (1.3- and 1.4-fold) and heterodimer FSH-Fc (5.9- and 7.1-fold) and remained elevated for >12 days after treatment with heterodimer FSH-Fc. CONCLUSIONS: We have shown that FSH-Fc fusion proteins have increased stability in blood and improved bioactivity in vivo, and that heterodimer FSH-Fc is more active in rats and monkeys than single chain FSH-Fc. These data suggest that Fc fusion proteins offer the potential for oral and pulmonary delivery of FSH. PMID- 15817591 TI - Laboratory safety during assisted reproduction in patients with blood-borne viruses. AB - For couples where one or both partners are infected with human immunodeficiency virus or hepatitis C, the doors to receiving fertility care are opening as a result of better antiviral medication, better long-term prognosis and consequent changes in attitude. In line with this, fertility centres electing to treat couples with blood-borne viral (BBV) infection need to re-examine their policies and procedures to ensure the safety of their staff and both non-infected and infected patients during assisted reproduction treatments. At a time when the European Tissue Directive aims to introduce quality standards for assisted reproduction throughout Europe, we highlight the risks involved when treating patients with known BBV infections and argue that safety cannot be met with any certainty unless samples from such patients are handled within a separate high security laboratory or laboratory area, technically adapted to ensure minimal cross-contamination risk to uninfected gametes and embryos. PMID- 15817592 TI - Conservative treatment in epithelial ovarian cancer: results of a multicentre study of the GCCLCC (Groupe des Chirurgiens de Centre de Lutte Contre le Cancer) and SFOG (Societe Francaise d'Oncologie Gynecologique). AB - BACKGROUND: Results of conservative management of epithelial ovarian cancer (EOC) remain controversial in the literature. The aim of this study was to assess the clinical outcomes and fertility following fertility-sparing surgical management of EOC in a retrospective multicentre study. METHODS: A multicentre retrospective study was performed by members of two French groups. Six inclusion criteria were defined: (i) Histological review by the same pathologist; (ii) age < or =40 years; (iii) conservative management; (iv) complete peritoneal staging; (v) delivery of a platinum-based chemotherapy in stage > or = IC; and (vi) follow-up >1 year. RESULTS: Thirty-four patients fulfilled the inclusion criteria: 30 had stage IA disease; three had stage IC and one had stage IIA. Eleven patients had recurrence: 10 patients had invasive disease and one had borderline recurrence. Among 10 patients with invasive recurrence, initial stage and grade were: stage IA G1, n = 1; stage IA G2, n = 4; stage IA G3, n = 1; and stage> or = IC, n = 4. All patients with stage > IA had recurrence. Ten pregnancies were observed in nine patients. CONCLUSION: Conservative surgery for patients with EOC could be considered in young patients with stage IA G1 disease. This procedure should not be performed in patients with FIGO stage > IA. PMID- 15817593 TI - Timing of CMF chemotherapy in combination with tamoxifen in postmenopausal women with breast cancer: role of endocrine responsiveness of the tumor. AB - BACKGROUND: Controversy persists about whether chemotherapy benefits all breast cancer patients. PATIENTS AND METHODS: In the International Breast Cancer Study Group (IBCSG) trial VII, 1212 postmenopausal patients with node-positive disease were randomized to receive tamoxifen for 5 years or tamoxifen plus three concurrent courses of cyclophosphamide, methotrexate and 5-fluorouracil ('classical' CMF) chemotherapy, either early, delayed or both. In IBCSG trial IX, 1669 postmenopausal patients with node-negative disease were randomized to receive either tamoxifen alone or three courses of adjuvant classical CMF prior to tamoxifen. Results were assessed according to estrogen receptor (ER) content of the primary tumor. RESULTS: For patients with node-positive, ER-positive disease, adding CMF either early, delayed or both reduced the risk of relapse by 21% (P=0.06), 26% (P=0.02) and 25% (P=0.02), respectively, compared with tamoxifen alone. There was no difference in disease-free survival when CMF was given prior to tamoxifen in patients with node-negative, ER-positive tumors. CONCLUSIONS: CMF given concurrently (early, delayed or both) with tamoxifen was more effective than tamoxifen alone for patients with node-positive, endocrine responsive breast cancer, supporting late administration of chemotherapy even after commencement of tamoxifen. In contrast, sequential CMF and tamoxifen for patients with node-negative, endocrine-responsive disease was ineffective. PMID- 15817594 TI - Adjuvant treatment for elderly patients with stage III colon cancer in the southern Netherlands is affected by socioeconomic status, gender, and comorbidity. AB - BACKGROUND: Adjuvant 5-fluorouracil-based chemotherapy significantly decreases mortality among patients with stage III colon cancer, but is less prescribed with rising age. We were interested in the pattern of adjuvant treatment and possible effects on survival among elderly patients. PATIENTS AND METHODS: All resected patients aged 65-79 with stage III colon carcinoma, diagnosed between 1995 and 2001 in the Comprehensive Cancer Centre South registry area in the Netherlands were included (n=577). We examined determinants of receipt of adjuvant chemotherapy and their relation to survival. RESULTS: The proportion of elderly patients receiving adjuvant chemotherapy increased from 19% in 1995 to 50% in 2001, but a large inter-hospital variation remained. In a multivariable analysis, females [odds ratio (OR) 0.5, P=0.006], patients with comorbidity [OR 0.5, P=0.005], and patients with a low socioeconomic status [OR 0.5, P=0.02] received less adjuvant therapy. Between 1995 and 2001 survival of elderly patients improved (hazard ratio 0.8, P=0.04). CONCLUSION: Although an increasing proportion of elderly patients with colon cancer are treated with adjuvant chemotherapy, many elderly patients still do not receive this treatment. As expected, receipt of adjuvant treatment decreased in the presence of comorbidity, but the clinical rationale for undertreatment of women and patients with low socioeconomic status is not clear. PMID- 15817595 TI - The influence of letrozole on serum lipid concentrations in postmenopausal women with primary breast cancer who have completed 5 years of adjuvant tamoxifen (NCIC CTG MA.17L). AB - BACKGROUND: The purpose of this study was to evaluate changes in serum lipid parameters {cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides and lipoprotein(a) [Lp(a)]}, in postmenopausal women receiving letrozole or placebo after adjuvant tamoxifen for early stage breast cancer (NCIC CTG MA.17L). PATIENTS AND METHODS: MA.17L is a substudy of MA.17, a randomized, double-blind, placebo-controlled trial of letrozole 2.5 mg taken daily for 5 years in postmenopausal women with primary breast cancer completing approximately 5 years of prior adjuvant tamoxifen. Patients consenting to participate in this companion study had blood drawn and lipid parameters (total cholesterol, HDL cholesterol, LDL cholesterol, Lp(a), triglycerides) evaluated at baseline, 6 months, 12 months and yearly thereafter until completion of protocol therapy. It was required that women be non hyperlipidemic and not taking lipid-lowering drugs at time of entry on this trial. RESULTS: Three hundred and forty seven women were enrolled in the study. The letrozole and the placebo groups demonstrated marginally significant differences in the percentage change from baseline in HDL cholesterol at 6 months (P=0.049), in LDL cholesterol at 12 months (P=0.033) and triglycerides at 24 months (P=0.036). All comparisons of lipid parameters at other time points were not significantly different between the two treatment groups. No statistically significant differences in the number of patients exceeding the thresholds defined for the lipid parameters were found between the two treatment groups. CONCLUSIONS: The MA.17 trial demonstrated a significant improvement in disease free survival with the use of letrozole as extended adjuvant therapy post tamoxifen. Results from this study suggests that letrozole does not significantly alter serum cholesterol, HDL cholesterol, LDL cholesterol, triglycerides or Lp(a) in non-hyperlidiemic postmenopausal women with primary breast cancer treated up to 36 months following at least 5 years of adjuvant tamoxifen therapy. These findings further support the tolerability of extended adjuvant letrozole in postmenopausal women following standard tamoxifen therapy. PMID- 15817596 TI - An implantable drug delivery system (IDDS) for refractory cancer pain provides sustained pain control, less drug-related toxicity, and possibly better survival compared with comprehensive medical management (CMM). AB - BACKGROUND: The randomized clinical trial of implantable drug delivery systems (IDDS) plus comprehensive medical management (CMM) versus CMM alone showed better clinical success at 4 weeks for IDDS patients. This 'as treated' analysis assessed if improvements in pain control, drug toxicity and survival were maintained over time. PATIENTS AND METHODS: We compared those who received IDDS with those who did not receive IDDS (non-IDDS). All patients had Visual Analogue Scores (VAS) for pain > or =5/10 on at least 200 mg morphine or equivalent daily. RESULTS: At 4 weeks, 46 of 52 (88.5%) IDDS patients achieved clinical success compared with 65 of 91 (71.4%; P=0.02) non-IDDS patients, and more often achieved > or =20% reduction in both pain VAS and toxicity [35 of 52 (67.3%) versus 33 of 91 patients (36.3%); P=0.0003]. By 12 weeks, 47 of 57 (82.5%) IDDS patients had clinical success compared with 35 of 45 (77.8%; P=0.55) non-IDDS patients, and more often had a > or =20% reduction in both pain VAS and toxicity [33 of 57 (57.9%) versus 15 of 45 patients (33.3%); P=0.01]. At 12 weeks the IDDS VAS pain scores decreased from 7.81 to 3.89 (47% reduction) compared with 7.21 to 4.53 for non-IDDS patients (42% reduction; P=0.23). The 12 week drug toxicity scores for IDDS patients decreased from 6.68 to 2.30 (66% reduction), and for non-IDDS patients from 6.73 to 4.13 (37% reduction; P=0.01). All individual drug toxicities improved with IDDS at both 4 and 12 weeks. At 6 months, only 32% of the group randomized to CMM and who did not cross over to IDDS were alive, compared with 52%-59% for patients in those groups who received IDDS. CONCLUSIONS: IDDS improved clinical success, reduced pain scores, relieved most toxicity of pain control drugs, and was associated with increased survival for the duration of this 6 month trial. PMID- 15817597 TI - Oxaliplatin combined with irinotecan and 5-fluorouracil/leucovorin (OCFL) in metastatic colorectal cancer: a phase I-II study. AB - BACKGROUND: A phase I-II multicenter trial was conducted to define the maximal tolerated dose and describe the activity of an OCFL combination using oxaliplatin (OHP), irinotecan (CPT-11) and 5-fluorouracil (FU)/leucovorin (LV) in metastatic colorectal cancer (CRC). PATIENTS AND METHODS: CRC patients not pretreated with palliative chemotherapy, with performance status < or =1 and adequate haematological, kidney and liver function, were eligible. Treatment consisted in weekly 24-h infusion 5-FU (2300 mg/m(2))/LV (30 mg) and alternating OHP (70-85 mg/m(2), days 1 and 15) and CPT-11 (80-140 mg/m(2), days 8 and 22) repeated every 5 weeks. OHP and CPT-11 were escalated in cohorts of three to six patients. RESULTS: Thirty patients received a median of five cycles. Dose-limiting toxicity occurred at dose level 3, and the recommended dose was OHP 70 mg/m(2), CPT-11 100 mg/m(2), LV 30 mg and 5-FU 2300 mg/m(2)/24 h. Grade > or =3 toxicities were diarrhea 23%, neutropenia 20%, fatigue 7%, and neurologic 7%. Two febrile neutropenia episodes (one fatal) were recorded. Among 28 patients with measurable disease (90%), we observed two complete and 20 partial responses; overall RR was 78% (95% CI, 59% to 92%). Median time to progression and overall survival were 9.5 and 25.4 months, respectively. Seven patients underwent liver metastases resection. CONCLUSION: OCFL is an overall well tolerated regimen with very high efficacy, which makes it most suitable for tumour control before surgery of metastatic disease. PMID- 15817598 TI - Tobacco dependence in the general population in Italy. AB - BACKGROUND: Measures of tobacco dependence are mainly used in the clinical setting, but limited information is available on tobacco dependence on a population level. MATERIALS AND METHODS: To obtain estimates of tobacco dependence on a population level, a six-item Fagerstrom questionnaire was used in two surveys, conducted in 2002-2003 on a sample of 6773 individuals aged 15 years or over, representative of the Italian adult population. RESULTS: Overall, 27.1% of Italian adults described themselves as current cigarette smokers (32.2% of men, 22.4% of women). Of all smokers, 42.8% were classified as very low dependent, 28.6% as low dependent, 11.0% as intermediate, 13.8% as high and only 3.8% as very high dependent. The proportions of very low/low dependent were 67.4% in men and 76.8% in women. Those of high/very high dependent smokers were 21.4% in men and 12.5% in women, but only 2.8% at age 15-17 years and 8.4% at age 18-24 years. Only 23% of smokers, moreover, found it difficult to avoid smoking in places where smoking was forbidden. CONCLUSIONS: The observation that over two thirds of smokers on a population level in Italy report low or very low dependence has useful implication for intervention on stopping smoking, particularly in the young, who appear to be low dependent. However, in this age group cessation rates were comparatively low. PMID- 15817599 TI - Predictors of quality of life in newly diagnosed melanoma and breast cancer patients. AB - BACKGROUND: Quality of life (QoL) is a multidimensional construct, and several psychosocial and medical factors can predict a patient's QoL. We investigated the impact of factors in the psychological stress processes on QoL 3 months after diagnosis of melanoma or breast cancer. PATIENTS AND METHODS: In consecutive melanoma and breast cancer patients with localized disease, cancer-specific social support, coping with cancer, personality trait anger expression, non cancer stress, sociodemographic variables, cancer and treatment variables, and QoL (evaluated as self-reported psychological and physical symptoms, self perceived QoL, and depression) were assessed with validated quantitative questionnaires. The associations between QoL and the other factors were investigated with multivariate methods. RESULTS: Only a few factors differed between melanoma and breast cancer, in terms of treatment modalities and gender. The amount of received social support was higher among breast cancer patients. The predicting power of psychological stress factors on all the QoL measures was strong, but differed between the treatment groups (surgery only versus adjuvant treatment). Psychosocial factors were the strongest predictors of QoL, not cancer type or treatment. Behavioral Escape-Avoidance coping was associated with worse QoL. However, non-cancer life stresses showed the strongest QoL-decreasing influence. QoL-enhancing factors, including social support, were seen clearly only within the breast cancer patients receiving adjuvant treatment. CONCLUSIONS: QoL of newly diagnosed cancer patients is highly associated with psychosocial factors. Non-cancer life stresses seem to be very important in the QoL of newly diagnosed cancer patients. Adjuvant treatment may comprise supportive psychosocial factors that enhance QoL in cancer. PMID- 15817600 TI - National and international nicotine dependence. PMID- 15817601 TI - Tailoring a tailored treatment: the importance of fine-tuning on the patient. PMID- 15817602 TI - High dose chemotherapy and autologous stem cell transplantation as adjuvant therapy for primary breast cancer patients with four or more lymph nodes involved: long-term results of an international randomised trial. AB - BACKGROUND: The purpose of this study was to assess whether a short course of anthracycline containing chemotherapy followed by high dose therapy with autologous stem-cell support improves disease-free and overall survival as compared with conventional, anthracycline containing chemotherapy, in patients with primary breast cancer and four or more histologically involved lymph nodes. PATIENTS AND METHODS: Two hundred and eighty one patients entered into a randomised clinical trial were allocated to receive standard, conventional treatment (5-fluorouracil, epirubicin and cyclophosphamide-FEC for six cycles) or FEC for three cycles followed by high dose therapy consisting of cyclophosphamide, thiotepa and carboplatin and stem cell rescue (HDT). To be eligible, patients had to be free of overt metastatic disease and be < or =60 years of age. Analyses were according to intention to treat. RESULTS: At a median follow up of 68 months, 118 patients have experienced a relapse or death from breast cancer (62 in the FEC followed by HDT arm and 56 in the conventional FEC arm) and a total of 100 patients have died (54 in the FEC followed by HDT arm and 46 in the conventional FEC arm). No significant difference was observed in relapse-free survival [hazard ratio 1.06, 95% CI 0.74-1.52, p = 0.76] or overall survival [hazard ratio 1.18, 95% CI 0.80-1.75, p = 0.40]. Five patients died from treatment related causes, three as a consequence of HDT and two in the conventional FEC arm. CONCLUSIONS: At the present time, no benefit has been observed from replacing three cycles of conventional chemotherapy with the HDT regimen described here. Patients should continue to receive conventional chemotherapy as adjuvant therapy for breast cancer. PMID- 15817603 TI - Management of anaemia in patients with breast cancer: role of epoetin. AB - Many patients with breast cancer suffer from anaemia, as a consequence of the disease itself or its treatment. Anaemia has a negative impact on treatment outcome and overall survival, and affects the quality of life (QoL) of patients with cancer. Previously, cancer-related anaemia was treated with blood transfusion, but this is inconvenient, offers only temporary improvement in haemoglobin (Hb) level and is associated with several risks. Consequently, blood transfusion is usually reserved for patients with severe anaemia (Hb levels <8 g/dl). Recombinant human erythropoietin (epoetin) is an effective and convenient treatment for cancer-related anaemia without the risks associated with red blood cell transfusion. Epoetin therapy effectively increases Hb levels, thereby reducing the need for emergency blood transfusion and improving the QoL of patients with anaemia and breast cancer. Epoetin beta is also effective for the prevention of anaemia and reduction of transfusion requirements in patients with a high risk of developing anaemia during chemotherapy. With the increased use of dose-intensified chemotherapy in an attempt to improve response rates, administration of epoetin to prevent anaemia could potentially benefit many patients with breast cancer. PMID- 15817604 TI - Randomized phase II trial of carboplatin versus paclitaxel and carboplatin in platinum-sensitive recurrent advanced ovarian carcinoma: a GEICO (Grupo Espanol de Investigacion en Cancer de Ovario) study. AB - BACKGROUND: The aim of this study was to determine whether the response rate for the paclitaxel-carboplatin combination is superior to carboplatin alone in the treatment of patients with platinum-sensitive recurrent ovarian carcinoma. PATIENTS AND METHODS: Patients with recurrent ovarian carcinoma, 6 months after treatment with a platinum-based regimen and with no more than two previous chemotherapy lines, were randomized to receive carboplatin area under the curve (AUC) 5 (arm A) or paclitaxel 175 mg/m(2) + carboplatin AUC 5 (arm B). The primary end point was objective response, following a 'pick up the winner' design. Secondary end points included time to progression (TTP), overall survival, tolerability and quality of life (QoL). RESULTS: Eighty-one patients were randomized and included in the intention-to-treat analysis. The response rate in arm B was 75.6% [26.8% complete response (CR) + 48.8% partial response (PR)] [95% confidence interval (CI) 59.7% to 87.6%] and 50% in arm A (20% CR + 30% PR) (95% CI 33.8% to 66.2%). No significant differences were observed in grade 3-4 hematological toxicity. Conversely, mucositis, myalgia/arthralgia and peripheral neurophaty were more frequent in arm B. Median TTP was 49.1 weeks in arm B (95% CI 36.9-61.3) and 33.7 weeks in arm A (95% CI 25.8-41.5). No significant differences were found in the QoL analysis. CONCLUSIONS: Paclitaxel carboplatin combination is a tolerable regimen with a higher response rate than carboplatin monotherapy in platinum-sensitive recurrent ovarian carcinoma. PMID- 15817605 TI - Immediate and mid-term outcomes of sirolimus-eluting stent implantation for chronic total occlusions. AB - AIMS: To evaluate the outcomes of sirolimus-eluting stent (SES) implantation for the treatment of chronic total occlusion (CTO). METHODS AND RESULTS: We identified 122 patients who underwent revascularization in CTO lesions with SES from April 2002 to April 2004 (SES group). A control group was composed of 259 consecutive patients with CTO lesions treated with bare metal stents (BMS) in the 24 months immediately before the introduction of SES (BMS group). At 6-month follow-up, the cumulative rate of major adverse cardiac events (MACE) was 16.4% in the SES group and 35.1% in the BMS group (P<0.001). The incidence of restenosis was 9.2% in the SES group and 33.3% in the BMS group (P<0.001). The need for revascularization in the SES group was significantly lower, both target lesion revascularization (7.4 vs. 26.3%, P<0.001) and target vessel revascularization (9.0 vs. 29.0%, P<0.001). BMS implantation (HR: 2.97; 95% CI: 1.80-4.89; P<0.001), lesion length (>20 mm) (HR: 2.02; 95% CI: 1.37-2.99; P=0.0004), and baseline reference vessel diameter (>2.8 mm) (HR: 0.62; 95% CI: 0.42-0.92; P=0.02) were identified as predictors of MACE during 6-month follow up. CONCLUSION: Compared with BMS, SES implantation in CTO lesions appears to be effective in reducing the incidence of restenosis and the need for revascularization at 6 months. PMID- 15817606 TI - Associations between cigarette smoking, pipe/cigar smoking, and smoking cessation, and haemostatic and inflammatory markers for cardiovascular disease. AB - AIMS: To examine the associations between cigarette smoking, pipe/cigar smoking, and years since quitting smoking, and inflammatory and haemostatic markers. METHODS AND RESULTS: A study in 2920 men aged 60-79 with no history of myocardial infarction, angina, stroke, or diabetes, and who were not on warfarin, from general practices in 24 British towns. After adjustment for other major cardiovascular risk factors, compared with never smokers, current cigarette smokers showed significantly higher levels of C-reactive protein (2.53 vs. 1.35 mg/L), white cell count (7.92 vs. 6.42 x 10(9)/L), and fibrinogen (3.51 vs. 3.13 g/L). They also showed higher levels of haematocrit, blood and plasma viscosity, tissue plasminogen activator antigen, and fibrin D-dimer, and lower levels of albumin. Primary pipe/cigar smokers showed levels similar to never smokers. Ex cigarette smokers and secondary pipe/cigar smokers showed intermediate levels although secondary pipe/cigar smokers showed higher odds of having elevated white cell count and fibrinogen than ex-cigarette smokers. Most inflammatory and haemostatic levels improved within 5 years of smoking cessation but took over 20 years to revert to levels of never smokers. CONCLUSION: These findings suggest that activation of inflammation and haemostasis may be potential mechanisms by which cigarette and pipe/cigar smoking increase cardiovascular risk. PMID- 15817607 TI - Non-invasive assessment of myocardial ischaemia using new real-time three dimensional dobutamine stress echocardiography: comparison with conventional two dimensional methods. AB - AIMS: Although two-dimensional-dobutamine stress echocardiography (2D-DSE) is useful for the diagnosis of myocardial ischaemia, it requires the acquisition of multiple cross-sections at each stage. The introduction of new real-time three dimensional echocardiography (RT3DE) offers rapid acquisition and 3D display of the entire left ventricle (LV). The purpose was to evaluate real-time three dimensional-dobutamine stress echocardiography (RT3D-DSE) for the diagnosis of ischaemia using exercise (201)Tl single-photon emission computed tomography (SPECT) as the reference standard, in comparison with 2D-DSE. METHODS AND RESULTS: We performed DSE in 56 consecutive patients who had undergone SPECT because of suspected ischaemia. 3D images by RT3DE were acquired from the apical window after the acquisition of cross-sectional images at every stage of 2D-DSE. Wall motion analysis in RT3DE was performed from anatomical images by cropping the acquired full volume data sets. Mean scanning time for adequate image acquisition at peak stress by RT3D-DSE was shorter than that by 2D-DSE (29+/-4 vs. 68+/-6 s, P<0.0001). RT3DE provided adequate images at success rate of 92% at rest and 89% at peak stress, whereas two-dimensional echocardiography did at 94 and 90%, respectively. The sensitivity, specificity, and accuracy of RT3D-DSE for the detection of coronary artery disease are 86, 80, and 82%, respectively. Those of 2D-DSE are 86, 83, and 84%, respectively. There were no significant differences in the sensitivity, specificity, and accuracy between these two methods (P=1.000). CONCLUSION: RT3D-DSE offers rapid and simple acquisition of the entire LV wall motion and provides feasible and accurate assessment of myocardial ischaemia. PMID- 15817608 TI - The p53 story: layers of complexity. AB - This report is a summary of the presentations at the 12th International p53 Workshop, held in November 2004, Dunedin, New Zealand. The Convenor of the organizing committee was Antony Braithwaite, University of Otago, Dunedin. Janice Royds and Paul Jackson were also members of the organizing committee. There were 61 oral presentations and 101 posters. PMID- 15817609 TI - Significant increase in risk of gastroesophageal cancer is associated with interaction between promoter polymorphisms in thymidylate synthase and serum folate status. AB - Thymidylate synthase (TS) catalyzes the 5,10-methylene-tetrahydrofolate-mediated conversion of deoxyuridine monophosphate to deoxythymydine monophosphate, a nucleotide required for DNA synthesis and repair. The impaired TS expression has been shown to be related to 28 bp tandem repeats and a G-->C SNP in the 5'-UTR of TS. Folate deficiency has been demonstrated to play a role in gastroesophageal carcinogenesis. This case-control study was to examine the hypothesis that the TS polymorphisms, alone or in combination with serum folate status, may confer susceptibility of the hosts to gastroesophageal cancer. We analyzed TS genotype and serum folate concentration in 324 patients with esophageal squamous cell carcinoma (ESCC), 231 patients with gastric cardia adenocarcinoma (GCA) and 492 controls. It was found that compared with the normal expression TS genotype, the low expression TS genotype alone was significantly associated with increased risk of ESCC [adjusted odds ratio (OR) 1.47; 95% confidence interval (CI) 1.03-2.10] but not GCA (OR=0.98, 95% CI=0.68-1.40). More importantly, a significant interaction between the TS polymorphisms and serum folate status in risk of ESCC and GCA was observed. Among subjects with low serum folate concentration (<3 ng/ml), the ORs of ESCC and GCA for the low expression genotype were 22.63 (95% CI=10.44-49.05) and 4.08 (95% CI=1.94-8.59), which were greater than respective 9.97 (95% CI=5.67-17.53) and 1.88 (95% CI=1.18-3.24) for the normal expression genotype (P=0.002 and 0.029). These results suggest an important role for folate deficiency and impaired TS activity in the etiology of ESCC and GCA. PMID- 15817610 TI - Associations between apoE genotype and colon and rectal cancer. AB - Apolipoprotein E (apoE) plays a major role in the metabolism of bile acids, cholesterol and triglycerides, and has recently been proposed as being involved in the carcinogenic process. Given the potential role of bile acids in colorectal cancer etiology, it is reasonable that colorectal cancer risk might be modified by apoE genotype. We used data collected from a case-control study of colon cancer (n=1556 cases and 1948 controls) and rectal cancer (n=777 cases and 988 controls). The absence of an e3 apoE allele significantly increased the risk of colon cancer (OR=1.37 95% CI 1.00-1.87), particularly among those diagnosed when older than 64 years (OR=1.88 95% CI 1.17-3.04; P interaction between age and apoE genotype equal to 0.05). A significant three-way interaction was detected for family history of colorectal cancer, age at diagnosis and apoE genotype (P = 0.05), in those diagnosed when older, not having an e3 allele and having a significantly increased risk of colon cancer with family history of colorectal cancer (OR=3.93 95% CI 1.23-12.6). This was compared with the risk associated with family history of colorectal cancer among those diagnosed when older, with an e3 allele of 1.61 (95% CI 1.17-2.23) or those diagnosed when younger without an e3 allele (OR=2.40 95% CI 0.56-10.3). Among those diagnosed when older than 64 years, associations of BMI and prudent diet with colon cancer were stronger among individuals without an e3 allele, although the P for interaction was not significant. We did not detect any significant associations between apoE genotype and rectal cancer, survival after diagnosis with colorectal cancer, stage of disease at diagnosis or type of tumor mutation. These findings suggest those apoE genotypes that do not include the e3 allele, the same genotypes that are associated with increased risk of coronary heart disease, may influence development of colon cancer among those who are older at diagnosis. PMID- 15817611 TI - Administration of green tea or caffeine enhances the disappearance of UVB-induced patches of mutant p53 positive epidermal cells in SKH-1 mice. AB - Irradiation of female SKH-1 hairless mice with UVB (30 mJ/cm2) twice a week for 10-20 weeks resulted in the formation of a large number of cellular patches (>8 adjacent cells/patch) that are recognized with an antibody (Pab240) which recognizes mutated but not wild-type p53 protein. These patches are not recognized by an antibody (Pab1620) to wild-type p53 protein. The patches, which are considered putative early cellular markers of the beginning of tumor formation, started appearing after 4-6 weeks of UVB treatment, and multiple patches were observed after treatment for 10 weeks. The number and size of the patches increased progressively with continued UVB treatment. Discontinuation of UVB for 4 weeks resulted in an 80-90% decrease in the number of these patches. The number of the remaining patches did not decrease any further but remained relatively constant for at least 4-9 weeks. Oral administration of green tea (6 mg tea solids/ml) or caffeine (0.4 mg/ml) as the sole source of drinking fluid during irradiation with UVB, twice a week for 20 weeks, inhibited UVB-induced formation of mutant p53 positive patches by approximately 40%. Oral administration of green tea (6 mg tea solids/ml) as the sole source of drinking fluid or topical applications of caffeine (6.2 micromol) once a day 5 days a week starting immediately after discontinuation of UVB treatment enhanced the rate and extent of disappearance of the mutant p53-positive patches. Topical applications of caffeine to the dorsal skin of mice pretreated with UVB for 20 weeks resulted in enhanced apoptosis selectively in focal basal cell hyperplastic areas of the epidermis (putative precancerous lesions), but not in areas of the epidermis that only had diffuse hyperplasia. Our studies indicate that the chemopreventive effect of caffeine or green tea may occur by a proapoptotic effect preferentially in early precancerous lesions. PMID- 15817612 TI - MnSOD inhibits proline oxidase-induced apoptosis in colorectal cancer cells. AB - Proline oxidase (POX), localized on inner mitochondrial membranes, is encoded by a p53-induced gene and metabolically participates in p53-induced apoptosis. Previously, we showed that POX catalyzed the generation of reactive oxygen species (ROS). We and others have demonstrated that overexpression of POX, independent of p53, causes apoptotic cell death in a variety of cancer cells. But a necessary role for ROS remains uncertain. Therefore, we asked whether superoxide dismutases (SOD) and catalase (CAT), important antioxidant enzymes, might interfere with the POX-dependent induction of apoptosis. In this study, we used DLD-1 colorectal cancer cells stably transfected with the POX gene under the control of a tetracycline-inducible promoter. When doxycycline was removed from the culture medium and the expression of POX was induced, apoptotic cell death was initiated. To examine the importance of the ROS-dependent component of the pathway, we infected DLD-1 POX cells with recombinant adenoviruses containing MnSOD, CuZnSOD, CAT or varying combinations of these adenoviruses followed by induced expression of POX. The expression of MnSOD inhibited POX-induced apoptosis, but others did not. Mechanistically, mitochondria-localized MnSOD dramatically reduced the release of cytochrome c to cytosol by POX. Compared with control cells, MnSOD-expressing DLD-1 POX cells generated a higher concentration of H2O2 owing to dismutation of superoxide radicals, which was elevated by POX. Thus, these data further suggest that the generation of superoxide radicals plays a crucial role in POX-induced apoptosis and the process is partially blocked by MnSOD. PMID- 15817613 TI - Hemoglobin adducts, urinary metabolites and health effects in 2,4,6 trinitrotoluene exposed workers. AB - 2,4,6-Trinitrotoluene (TNT) is an important occupational and environmental pollutant. In TNT exposed humans, the notable toxic manifestations have included aplastic anemia, toxic hepatitis, cataract, hepatomegaly and liver cancer. Therefore, we developed methods to biomonitor workers exposed to TNT. The workers were employed in a typical ammunition factory in China. The controls were recruited from the same factory. We determined hemoglobin (Hb) adducts and urine metabolites of TNT. Hb-adducts of TNT, 4-amino-2,6-dinitrotoluene (4ADNT) and 2 amino-4,6-dinitrotoluene (2ADNT), and the urine metabolites of TNT, 4ADNT and 2ADNT were found in all the workers and in a few controls. 4ADNT was the main product. Although the levels of 2ADNT correlated well with 4ADNT, 2ADNT was not found in all the samples. Therefore, 4ADNT was the best marker of exposure for Hb adducts and urine metabolites. The levels of the urine metabolites and Hb-adducts were related to the health status of the workers. The Hb-adduct 4ADNT was statistically significantly associated with risk of hepatomegaly, splenomegaly and cataract. The odds ratio (OR) for cataract, splenomegaly and hepatomegaly were 6.4 [95% confidence interval (CI) = 1.4-29.6], 9.6 (1.1-85.3) and 7.6 (1.3 43.7), respectively. No correlation was found between urine metabolites and health effects. These results were tested for confounding factors like age, workyears, smoker status, smoke years, cigarettes per day and hepatitis B status using stepwise forward logistic regression analysis. In the case of splenomegaly, hepatitis B status is a confounder. In the case of cataract, age is a confounder. The Hb-adduct, 4ADNT, is a good biomarker of exposure and biomarker of biological effect. PMID- 15817614 TI - Prostate cancer risk and DNA damage: translational significance of selenium supplementation in a canine model. AB - Daily supplementation with the essential trace mineral selenium significantly reduced prostate cancer risk in men in the Nutritional Prevention of Cancer Trial. However, the optimal intake of selenium for prostate cancer prevention is unknown. We hypothesized that selenium significantly regulates the extent of genotoxic damage within the aging prostate and that the relationship between dietary selenium intake and DNA damage is non-linear, i.e. more selenium is not necessarily better. To test this hypothesis, we conducted a randomized feeding trial in which 49 elderly beagle dogs (physiologically equivalent to 62-69-year old men) received nutritionally adequate or supranutritional levels of selenium for 7 months, in order to mimic the range of dietary selenium intake of men in the United States. Our results demonstrate an intriguing U-shaped dose-response relationship between selenium status (toenail selenium concentration) and the extent of DNA damage (alkaline Comet assay) within the prostate. Further, we demonstrate that the concentration of selenium that minimizes DNA damage in the aging dog prostate remarkably parallels the selenium concentration in men that minimizes prostate cancer risk. By studying elderly dogs, the only non-human animal model of spontaneous prostate cancer, we have established a new approach to bridge the gap between laboratory and human studies that can be used to select the appropriate dose of anticancer agents for large-scale human cancer prevention trials. From the U-shaped dose-response, it follows that not all men will necessarily benefit from increasing their selenium intake and that measurement of baseline nutrient status should be required for all individuals in prevention trials to avoid oversupplementation. PMID- 15817615 TI - Guest editorial: what do prospective randomized clinical trials tell us about the treatment of class II malocclusions? A personal viewpoint. AB - The prospective randomized clinical trial (RCT) is seen by many to be the 'gold standard' for analysing treatment outcome and the only valid source of clinical data. In orthodontics, most RCTs have been designed to resolve the controversy surrounding the ability of functional appliances to significantly modify dentofacial growth. Given the variability in the timing, magnitude and duration of pubertal dentofacial growth, differing levels of motivation and patient compliance, the inherent inaccuracy of cephalometry and the questionable validity of the measurements themselves used to quantitate change, it is not surprising that the conclusions have not been as clear-cut as anticipated. Unlike a laboratory experiment, in which it is possible to limit the differences between experimental and control groups to the single factor being investigated, in a clinical trial an orthodontic appliance is just one of several variables affecting the outcome. Furthermore, RCTs are expensive and time-consuming; by the time the end-point of the study has been reached, the appliance may no longer be in widespread use, the fate of more than one recent well-publicized RCT. What RCTs have shown is that functional appliances such as the Herbst and twin block, based on the principle of 'jumping the bite', are more effective at modifying dentofacial growth and reducing overjets than headgear and more passive appliances such as the Andresen activator and its variants. However, if one asks whether RCTs have achieved their objective, or provided knowledge not previously available from retrospective studies or animal experimentation, then the answer would have to be no; it is also hard to justify the cost. What is particularly interesting is that knowledge based on years of clinical experience has been disregarded and then announced as if it was something completely new. PMID- 15817616 TI - Permissible limit for mandibular expansion. AB - In recent years, mandibular expansion has been increasingly performed in conjunction with orthodontic treatment. Lateral tipping of the molars associated with mandibular expansion should, however, be considered, because excessive expansion may result in excessive buccal tooth inclination, which may disturb the occlusal relationship. This study was conducted to quantitatively clarify molar movement during mandibular expansion using the Schwarz appliance to determine the permissible limit of mandibular expansion as a clinical index for inclination movement. Inclinations in the masticatory surface of the first molar and intermolar width were measured before expansion (T1), after expansion (T2), and before edgewise treatment (T3). Lower plaster models from 29 subjects treated with expansion plates were used and compared with models from 11 control subjects with normal occlusion. The average treatment change (T1-T2) in intermolar width was 5.42 mm (standard deviation 1.98), and the average angle of buccal tooth inclination was 10.16 degrees (standard deviation 3.83). No significant correlation was found between age prior to treatment and the treatment period when they were compared with the intermolar width increments and inclination angles. There was a significant positive correlation between retention duration and the amount of expansion. The regression coefficient of the angle of buccal tooth inclination during expansion to the increment of the intermolar width was approximately 0.2. This means that 1 mm of expansion is accompanied by 5 degrees of molar lateral tipping. This coefficient is clinically useful for estimating the permissible limit for mandibular expansion. PMID- 15817617 TI - Changes in clinical crown height as a result of transverse expansion of the maxilla in adults. AB - The risk of developing bony dehiscence and gingival recession may lead clinicians to prefer extraction to expansion in borderline cases. The purpose of this research was to compare changes in clinical crown height that occur at the buccal aspect of the maxillary lateral teeth of adult patients in which the transverse dimension was increased with those occurring if no expansion had been performed. Secondly it was the intention to identify factors increasing the risk of development of gingival recessions. The material comprised the pre- and post treatment study casts from two groups of 50 adult patients. In one group an average transverse expansion of 3 mm was performed while in the other no change in the arch width was generated during treatment. Clinical crown heights of the two groups before and after treatment were compared with a Student's t-test and correlation analysis was used to determine whether any demographic or treatment related parameters could be applied to predict an increase in clinical crown height. The increase in transarch width was greater in males (2.4-3.4 mm) than in females (1.8-2.5 mm), and greater at the level of the premolars than at the molars. No significant increase in buccal crown height could be identified. The increase in width was, however, related to buccal tipping as a positive correlation was found between the amount of expansion and tipping (P < 0.01). No detrimental effect of slow maxillary expansion could be demonstrated. PMID- 15817618 TI - The influence of mandibular prominence on facial attractiveness. AB - This study examined the attractiveness of facial profiles. One hundred and two social science students (28 males and 74 females) rated the attractiveness of a series of silhouettes with normal, Class II or Class III profiles. A random sequence of 10 images included an image with the Eastman normal SNB value of 78 degrees, and images with SNB values of 2.5, 5, 7.5 and 10 degrees above and below normal. A duplicate image in each sequence was used to assess reproducibility. The participants scored the attractiveness of each image and also indicated whether they would seek treatment if each image was their own profile. The profile with the normal SNB angle of 78 degrees was rated as the most attractive. Attractiveness scores reduced as the mandibular profile diverged from the normal SNB value. The +5 degree profile (SNB = 83 degrees) was rated as significantly more attractive than the -5 degree profile (SNB = 73 degrees; P = 0.004). No other significant differences between the scores for Class II and Class III profile pairs of equal severity were found. At 10 degrees below the normal SNB (Class II), 74 per cent of the sample would elect to have treatment, while 78 per cent would elect to have treatment at 10 degrees above the normal SNB (Class III). PMID- 15817619 TI - Attractiveness ratings of anterior open bites and reverse overjets using the aesthetic component of the Index of Orthodontic Treatment Need. AB - This study was carried out to determine the level of attractiveness of anterior open bites (AOB) and reverse overjets of varying severity. A sample of 180 non dental students (101 females and 79 males; average age 20 +/- 0.75 years) and 45 dental professionals (12 females and 33 males; average age 35.5 +/- 5.07 years) was asked to complete a questionnaire to rate the level of attractiveness of AOB and reverse overjets of varying severity using the aesthetic component (AC) of the Index of Orthodontic Treatment Need (IOTN). Photographs of mild and severe AOB and reverse overjets were projected onto a white screen in a lecture theatre (15 seconds each with a 5 second interval between images). The participants were asked to record the AC grade at which they thought the projected picture of the AOB or reverse overjets had similar attractiveness. Dental awareness of non dental students was determined by asking them their opinion on how important it was to have straight teeth (very unimportant, unimportant, important, very important), about their personal or close family members' orthodontic experiences and if they thought they were in need of any orthodontic treatment. A chi-square test was applied to record any differences between sexes and between the different groups. Backward stepwise linear regression analysis was used to determine the relationship between the students' ratings of the photographs and their dental awareness. The majority of non-dental students rated a mild AOB (93 per cent) and mild reverse overjet (96 per cent) to be aesthetically acceptable. A mild AOB and mild reverse overjet were found to be acceptable by 40 and 58 per cent of dental professionals, respectively. The differences in the ratings between dental professionals and non-dental students were significant at P < 0.001.A severe AOB was considered unattractive by both students and dental professionals. However, dental professionals rated it at the more unattractive end of the scale (P < 0.001). A severe reverse overjet was rated by the majority of the subjects as aesthetically unacceptable (85 per cent of the non-dental students and 78 per cent of the dental professionals). PMID- 15817620 TI - Self-perceived orthodontic treatment need and culturally related differences among adolescents in Sweden. AB - The aim of this investigation was to compare Swedish and immigrant groups of 12- and 13-year-old boys and girls with respect to: (1) self-perceived need for and attitude to orthodontic treatment, (2) attitude to own teeth and general appearance, (3) behaviour pattern and psychosocial functioning, and (4) self perceived need for orthodontic treatment in relation to psychosocial functioning. The subjects comprised 508 students, aged 12 and 13 years, living in Sweden, who were grouped according to nationality: (A) both parents born in Sweden (139 girls and 131 boys); at least one parent born in: (B) eastern Europe (27 girls and 34 boys); (C) Asia (66 girls and 61 boys) and (D) other countries (23 girls and 27 boys). Each student answered a questionnaire in the classroom. The questions concerned demographic data, self-perceived treatment need, attitude to orthodontic treatment, own teeth and general appearance, behaviour pattern and psychosocial functioning. The results showed that, on average, 20 per cent of the students had a self-perceived treatment need, more girls than boys, 24 per cent of Swedes (A), 12 per cent from eastern Europe (B), 18 per cent from Asia (C) and 14 per cent from other countries (D). Seventy-two per cent of the Swedish students were prepared to undergo fixed appliance therapy, compared with 58 per cent of immigrant students. Nine per cent of the Swedish students considered their general appearance to be less favourable compared with 7 per cent of their peers (not significant). While some differences in behaviour pattern were observed, these did not seem to influence the well-being of the subjects. The conclusion is that perceived orthodontic treatment need is lower in immigrant students than in Swedish students. PMID- 15817621 TI - Attitudes towards orthodontic treatment: a comparison of treated and untreated subjects. AB - The aims of the present study were to evaluate treated and untreated subjects' attitudes towards orthodontic treatment and to examine possible determinants of these attitudes. It was hypothesized that orthodontically treated individuals would differ from untreated respondents in their attitude towards orthodontists and orthodontic treatment, and that female subjects would have a more positive attitude towards orthodontics than male subjects. Untreated individuals (n = 220) were used as a comparison group in the evaluation of orthodontic health care by previously treated subjects (n = 246). Two questionnaires were completed. The first, based on the Dental Attitude Questionnaire, contained 32 items about general attitude towards orthodontic treatment and was completed by both groups. The second questionnaire contained 46 negatively and positively based statements concerning different aspects of orthodontic treatment, and was completed by previously treated subjects. The reliability of both questionnaires was satisfactory. Previously treated subjects were found to have a significantly more positive attitude towards orthodontics than untreated subjects. The subject's attitude towards the relationship with the orthodontist, satisfaction with the treatment result and experiences with follow-up appointments predicted the general attitude towards orthodontics. Age, but not gender, was found to be a significant predictor for a subject's general attitude towards orthodontics. PMID- 15817622 TI - Does orthodontic treatment affect patients' and parents' anxiety levels? AB - The aims of this study were (1) to determine and compare the anxiety levels of two groups of patient and parents, (2) to identify possible gender differences between male and female subjects and (3) to evaluate any changes in anxiety levels after 1 year of treatment. The first group consisted of 40 subjects with a mean age of 15.6 +/- 1.2 years awaiting orthodontic treatment, plus one parent of each subject (mean age 43.4 +/- 2.3 years). The second group comprised 43 patients with a mean age of 16.0 +/- 1.1 years who had been undergoing treatment for a period of 1 year, plus one parent of each patient (mean age 41.0 +/- 1.9 years).Personal information forms and Spielberger's 'State and Trait Anxiety Inventory' (STAI) were applied to both groups. To compare the two groups and to determine the differences between males and females, independent-sample t-tests were used. Internal consistencies for the two scales of the STAI were evaluated with Cronbach's alpha coefficient. Trait anxiety levels of parents (51.05 +/- 5.1) and state anxiety levels of subjects (58.57 +/- 6.73) who were about to start orthodontic treatment were both high. The difference between the groups was statistically significant (P < 0.05). In patients who had undergone treatment for 1 year, the scores were found to be normal (43.28 +/- 5.91). However, their parents' high levels of trait anxiety remained unchanged (50.41 +/- 4.2). PMID- 15817623 TI - Evaluation of the aesthetic component of the Index of Orthodontic Treatment Need by Swedish orthodontists. AB - The aim of the present study was to evaluate if the majority of orthodontists in Sweden agree with the ranking of the photographs in the aesthetic component (AC) of the Index of Orthodontic Treatment Need (IOTN), and its treatment need classification. Ten separate colour photographs (originals from the AC) and two questionnaires were sent to 272 orthodontists in Sweden. Eighty-one per cent (219) returned one or both questionnaires. One hundred and ninety-four orthodontists answered the AC 'grading of dental attractiveness' and 217 the AC 'need for orthodontic treatment' (nine of the integrated questions had to be excluded because more than one alternative had been chosen). The aesthetic ranking by the orthodontists was compared with the original AC and the deviation was estimated. The results showed that the grading of photographs 2-9 varied greatly among the orthodontists, but the agreement was almost complete for photographs 1 and 10. The establishment of treatment need for each photograph had good agreement with the original index for photographs 1, 2, and 8-10, while the majority of the orthodontists considered that photographs 5 and 7 should be treated and that photograph 6 showed a borderline case. The participating Swedish orthodontists' aesthetic ranking of the photographs was: 1, 2, 3, 4, 6, 5, 7, 9, 8, and 10. Regarding treatment need, no need for treatment was set for photographs 1-4, borderline for photograph 6 and a need for treatment for photographs 5, 7-10. Further studies are needed to evaluate if laymen in Sweden make the same judgements as Swedish orthodontists. PMID- 15817624 TI - Differentiation of developmental and post-orthodontic white lesions using image analysis. AB - The aim of this study was to investigate differences in shape and size characteristics between developmental opacities and post-orthodontic white enamel lesions using computerized image analysis. Material, in the form of 35 mm slides, was obtained from the archive of photographic patient records in the orthodontic clinic at the Charles Clifford Dental Hospital. Images of 30 teeth with developmental white lesions and 30 teeth with post-orthodontic white lesions were selected using strict inclusion and exclusion criteria. The slides were converted to a digital format, coded, placed in a random order and analysed blindly using a computerized image analysis system by one clinician. After a 2 week interval, the images were recorded, placed in a new random order and the measurements repeated. The outcome measures were: area and luminance proportionality, and the shape of the perimeter line (expressed as the mathematical factor, roundness). Reproducibility was assessed by a paired samples t-test for systematic error and the intra-class correlation coefficient (ICC) for random error. Differences between groups were tested using the Mann-Whitney U-test for non-parametric data. Reproducibility was substantial for all measurements except for developmental white lesion roundness, which was moderate. There was a statistically significant difference between developmental white opacities and post-orthodontic white lesions for measurements of luminance intensity, proportionality (P = 0.002) and roundness (P = 0.001). Developmental white opacities had a higher luminance (i.e. were whiter) and the boundaries were more circular in shape than the post orthodontic lesions. Roundness is a useful measure when distinguishing developmental and post-orthodontic demineralization. PMID- 15817625 TI - Degree of conversion of two lingual retainer adhesives cured with different light sources. AB - The aim of this study was to evaluate the degree of conversion (DC) of two lingual retainer adhesives, Transbond Lingual Retainer (TLR) and Light Cure Retainer (LCR), cured with a fast halogen light, a plasma arc light and a light emitting diode (LED) at various curing times. A conventional halogen light served as the control. One hundred adhesive samples (five per group) were cured for 5, 10 or 15 seconds with an Optilux 501 (fast halogen light), for 3, 6 or 9 seconds with a Power Pac (plasma arc light), or for 10, 20 or 40 seconds with an Elipar Freelight (LED). Samples cured for 40 seconds with the conventional halogen lamp were used as the controls. Absorbance peaks were recorded using Fourier transform infrared (FT-IR) spectroscopy. DC values were calculated. Data were analysed using Kruskal-Wallis and Mann-Whitney U-tests. For the TLR, the highest DC values were achieved in 6 and 9 seconds with the plasma arc light. Curing with the fast halogen light for 15 seconds and with the LED for 40 seconds produced statistically similar DC values, but these were lower than those with the plasma arc light. All of these light exposures yielded a statistically significantly higher DC than 40 seconds of conventional halogen light curing. The highest DC value for the LCR was achieved in 15 seconds with the fast halogen light, then the plasma arc light curing for 6 seconds. These two combinations produced a statistically significantly higher DC when compared with the 40 seconds of conventional halogen light curing. The lowest DC for the LCR was achieved with 10 seconds of LED curing. The overall DC of the LCR was significantly higher than that of the TLR. The results suggest that a similar or higher DC than the control values could be achieved in 6-9 seconds by plasma arc curing, in 10-15 seconds by fast halogen curing or in 20 seconds by LED curing. PMID- 15817626 TI - The variable anchorage straight wire technique compared with the straight wire technique in deep overbite correction. AB - The aim of this retrospective study was to compare the treatment results and the number of required treatment appointments between the variable anchorage straight wire technique (VAST) and the straight wire (SW) technique. The subjects were 53 Caucasian patients of both sexes (25 males and 28 females, mean age 13.5 years at the start of treatment), with an overjet > or = 4 mm and an overbite > or = 3 mm. The patients were divided into four groups, VAST (n = 31) or SW (n = 22), extraction or non-extraction, and were treated by the same orthodontist. Variables from two lateral cephalograms obtained before and at the completion of active treatment, and the number of scheduled appointments were compared between the two techniques. The main difference between the two techniques was the bracket design. With the VAST, the bracket allowed both tipping and parallel movements with the possibility to combine double archwires. Due to the influence of the Begg technique, no extra-oral traction was needed in the VAST groups and Class II elastics were used at the start of treatment. Both techniques seemed to produce equal treatment results. However, in this study, it was shown that in deep overbite correction, the VAST required fewer scheduled appointments than the SW technique. PMID- 15817627 TI - Association between ectopic eruption of maxillary canines and first molars. AB - The purpose of this study was to elucidate a possible association between ectopic first molar eruption causing root resorption on the distal root of the primary maxillary second molar, and ectopic canine eruption causing root resorption on the permanent maxillary incisors. The subjects consisted of 30 patients, 22 females and eight males in the age range 8.3-15.0 years in whom root resorption of the permanent maxillary lateral and/or central incisor caused by the erupting permanent canine was diagnosed, and clinical and/or radiographic information concerning maxillary first molar eruption existed. It was found that of the 30 patients, seven (23.3 per cent) also had pathological root resorption of the second primary molar caused by ectopic molar eruption. It is suggested that patients with ectopic maxillary first molar eruption leading to pathological root resorption of the maxillary second primary molar are followed closely during the period of premolar and canine eruption, as the ectopic first molar could be an early warning of an increased risk of ectopic canine eruption leading to root resorption of the maxillary permanent incisors. PMID- 15817628 TI - Validation of speed of sound for the assessment of cortical bone maturity. AB - Bone changes its structural and physical properties during maturation. In order to validate ultrasound measurements with regard to their usefulness in assessing cortical bone maturation, speed of sound (SOS) data were compared with mechanical properties (elastic modulus, bending strength, and cortical surface hardness), density and water content. Thirty pig mandibles were selected from three different age groups. Rectangular specimens of the buccal cortical bone of the body of the mandible were prepared. SOS was measured with pulsed ultrasound at a frequency of 2 MHz in all three dimensions, bone mineral density (BMD) by quantitative computed tomography, breaking strength and apparent elastic modulus in a three-point bending test to failure, water content using the lyophilization technique, and micro-indentation hardness using a modified Vickers' technique. While SOS in all three directions, BMD, surface hardness, and bending strength increased significantly (P < 0.001), bone tissue water content decreased significantly (P < 0.001) with age. Changes in the elastic modulus were not significant. Changes in SOS in the antero-posterior and apico-occlusal directions can be partly explained by BMD. In a bucco-lingual direction the increase is inadequately explained by the physical parameters investigated, and has possibly to be attributed to structural differences. Maturation of the mandible implies changes in its architectural organization, in material composition, and in the mechanical properties of cortical bone. In vitro SOS measurements reflect different structural and physical properties that are all age dependent. It thus seems feasible that age-related changes in bone maturation could be monitored by SOS measurements. PMID- 15817629 TI - Clinical evaluation of the centre of resistance of the upper incisors during retraction. AB - The aim of this study was to evaluate the movement of anterior teeth during retraction with a force applied through the assumed centre of resistance (CRe).Twenty-two subjects with a Class I or II malocclusion were included. Each subject had the two upper first premolars extracted, resulting in a symmetrical extraction space of at least 3 mm between the upper laterals and canines. The force was applied through the assumed CRe, located 9 mm gingival to the lateral tooth bracket. To examine the type of anterior tooth movement, 10 parameters were measured. A Wilcoxon test was used to determine the differences between pre- and post-retraction values, and a Mann-Whitney U-test to determine the mean differences between groups. In spite of the force application through the CRe, tipping of the anterior teeth was observed in 19 subjects and parallel movement in three patients. Consequently, the subjects were divided into two groups according to the location of the centre of rotation (CRo). In group 1 (nine cases), the CRo was located coronal to the root apex, and in group 2 (13 cases), apical to the root apex. Both groups showed a significant decrease in inclination (P < 0.01) and posterior crown movement (P < 0.01 for group 1 and P < 0.001 for group 2) of the anterior teeth. A significant posterior movement of the root apex was observed in group 2 (P < 0.001). Significant differences were found between the groups for anterior tooth inclination (P < 0.05) and root apex movement (P < 0.001). The reasons for these differences could not be conclusively determined. Even though experimental studies provide information regarding CRe location, factors such as bone support, root morphology and incisor inclination should be taken into consideration. The observation of tooth movement occurring during treatment and changes in treatment mechanics would be helpful in obtaining desired tooth movement. PMID- 15817630 TI - Matrix metalloproteinase-1 and -8 in gingival crevicular fluid during orthodontic tooth movement: a pilot study during 1 month of follow-up after fixed appliance activation. AB - The role of matrix metalloproteinases (MMPs) in response to mechanical forces in orthodontic tooth movement has only been partially clarified. In the present in vivo pilot study, the presence, levels, and degree of activation of MMP-1 and -8 were measured daily for 1 month in gingival crevicular fluid (GCF) of patients treated with orthodontic fixed appliances. GCF samples were collected from five orthodontic patients and three controls from one upper or lower central incisor or from one upper canine before fixed appliance activation and every 24 hours for 1 month thereafter. The molecular forms and activation degrees of MMP-1 and -8 in GCF were analysed by Western blotting, and MMP-8 levels determined by immunofluorometric assay (IFMA).IFMA revealed, during the study period, on average 12-fold higher levels (56 +/- 50 versus 4.6 +/- 4 microg/l) of MMP-8 in orthodontic GCF than in control GCF. The MMP-8 levels in orthodontic GCF were lower than those detected in gingivitis and periodontitis GCF, but significantly higher than in control GCF. IFMA analysis was confirmed by Western blot analysis showing elevated MMP-8 levels from orthodontic GCF relative to control GCF. Forty one per cent of total MMP-8 immunoreactivities were high-molecular weight complexes (>100 kDa), 32 per cent in the 75 kDa pro-polymorphonuclear (PMN)-MMP-8 form, 14 per cent in the 60 kDa active-PMN-MMP-8 form, and 13 per cent in the 55 kDa fibroblast-type pro-MMP-8 form. In the GCF of orthodontic patients no MMP-1 immunoreactivities were detected. MMP-8 and -1 levels in the control GCF were low and not detectable. These results demonstrate that in vivo in human GCF, elevation and partial activation of multiple species of PMN- and fibroblast-type MMP-8 reflect periodontal remodelling during orthodontic tooth movement. PMID- 15817631 TI - Local recovery of Ca2+ release in rat ventricular myocytes. AB - Excitation-contraction coupling in the heart depends on the positive feedback process of Ca2+-induced Ca2+ release (CICR). While CICR provides for robust triggering of Ca2+ sparks, the mechanisms underlying their termination remain unknown. At present, it is unclear how a cluster of Ca2+ release channels (ryanodine receptors or RyRs) can be made to turn off when their activity is sustained by the Ca2+ release itself. We use a novel experimental approach to investigate indirectly this issue by exploring restitution of Ca2+ sparks. We exploit the fact that ryanodine can bind, nearly irreversibly, to an RyR subunit (monomer) and increase the open probability of the homotetrameric channel. By applying low concentrations of ryanodine to rat ventricular myocytes, we observe repeated activations of individual Ca2+ spark sites. Examination of these repetitive Ca2+ sparks reveals that spark amplitude recovers with a time constant of 91 ms whereas the sigmoidal recovery of triggering probability lags behind amplitude recovery by approximately 80 ms. We conclude that restitution of Ca2+ sparks depends on local refilling of SR stores after depletion and may also depend on another time-dependent process such as recovery from inactivation or a slow conformational change after rebinding of Ca2+ to SR regulatory proteins. PMID- 15817632 TI - The nitric oxide donor sodium nitroprusside stimulates the Na+-K+ pump in isolated rabbit cardiac myocytes. AB - Nitric oxide (NO) affects the membrane Na(+)-K(+) pump in a tissue-dependent manner. Stimulation of intrinsic pump activity, stimulation secondary to NO induced Na(+) influx into cells or inhibition has been reported. We used the whole-cell patch clamp technique to measure electrogenic Na(+)-K(+) pump current (I(p)) in rabbit ventricular myocytes. Myocytes were voltage clamped with wide tipped patch pipettes to achieve optimal perfusion of the intracellular compartment, and I(p) was identified as the shift in holding current induced by 100 microm ouabain. The NO donor sodium nitroprusside (SNP) in concentrations of 1, 10, 50 or 100 microm induced a significant increase in I(p) when the intracellular compartment was perfused with pipette solutions containing 10 mm Na(+), a concentration near physiological levels. SNP had no effect when the pump was near-maximally activated by 80 mm Na(+) in pipette solutions. Stimulation persisted in the absence of extracellular Na(+), indicating its independence of transmembrane Na(+) influx. The SNP-induced pump stimulation was abolished by inhibition of soluble guanylyl cyclase (sGC) with 1H-[1,2,4]oxadiazole[4,3 a]quinoxalin-1-one, by inhibition of protein kinase G (PKG) with KT-5823 or by inhibition of protein phosphatase with okadaic acid. Inclusion of the non hydrolysable cGMP analogue 8pCPT-cGMP, activated recombinant PKG or the sGC activator YC-1 in patch pipette filling solutions reproduced the SNP-induced pump stimulation. Pump stimulation induced by YC-1 was dependent on the Na(+) concentration but not the K(+) concentration in pipette filling solutions, suggesting an altered sensitivity of the Na(+)-K(+) pump to intracellular Na(+). PMID- 15817633 TI - Taurine activates excitatory non-synaptic glycine receptors on dopamine neurones in ventral tegmental area of young rats. AB - The physiological and pharmacological properties of taurine-induced responses were investigated in dopaminergic (DA) neurones from the ventral tegmental area (VTA) of young rats aged 1-13 postnatal days, either in acute brain slices or acutely dissociated neurones. When whole-cell responses were recorded from current-clamped neurones using the gramicidin-perforated technique, the application of taurine (0.01-30 mm) accelerated firings and induced membrane depolarization. In voltage-clamped neurones, taurine induced a current which was antagonized by strychnine and by picrotoxin, but not by bicuculline. In addition, taurine-induced current showed complete cross-desensitization with glycine activated currents but not with gamma-aminobutyric acid (GABA)-activated currents. Thus, taurine is a full agonist of the glycine receptors (GlyRs) in the VTA. Further studies found that taurine acted mainly on non-synaptic GlyRs. The application of 20 microm bicuculline abolished the spontaneous inhibitory post synaptic currents (IPSCs) in 40/45 neurones, and 93% of the evoked IPSCs. The addition of 1 microm strychnine completely eliminated the remaining IPSCs. These results suggest that GABAergic IPSCs predominate, and that functional glycinergic synapses are present in a subset of the VTA neurones. The application of 1 mum strychnine alone induced an outward current, suggesting that these neurones were exposed to tonically released taurine/glycine. In conclusion, by activating non synaptic GlyRs, taurine may act as an excitatory extra-synaptic neurotransmitter in the VTA during early development. PMID- 15817634 TI - Critical amino acid residues involved in the electrogenic sodium-bicarbonate cotransporter kNBC1-mediated transport. AB - We have previously reported a topological model of the electrogenic Na(+)-HCO(3)( ) cotransporter (NBC1) in which the cotransporter spans the plasma membrane 10 times with N- and C-termini localized intracellularly. An analysis of conserved amino acid residues among members of the SLC4 superfamily in both the transmembrane segments (TMs) and intracellular/extracellular loops (ILs/ELs) provided the basis for the mutagenesis approach taken in the present study to determine amino acids involved in NBC1-mediated ion transport. Using large-scale mutagenesis, acidic and basic amino acids putatively involved in ion transport mediated by the predominant variant of NBC1 expressed in the kidney (kNBC1) were mutated to neutral and/or oppositely charged amino acids. All mutant kNBC1 cotransporters were expressed in HEK-293T cells and the Na(+)-dependent base flux of the mutants was determined using intracellular pH measurements with 2',7'-bis (carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Critical glutamate, aspartate, lysine, arginine and histidine residues in ILs/ELs and TMs were detected that were essential for kNBC1-mediated Na(+)-dependent base transport. In addition, critical phenylalanine, serine, tyrosine, threonine and alanine residues in TMs and ILs/ELs were detected. Furthermore, several amino acid residues in ILs/ELs and TMs were shown to be essential for membrane targeting. The data demonstrate asymmetry of distribution of kNBC1 charged amino acids involved in ion recognition in putative outward-facing and inward-facing conformations. A model summarizing key amino acid residues involved in kNBC1-mediated ion transport is presented. PMID- 15817635 TI - Opposing muscarinic and nicotinic modulation of hypoglossal motor output to genioglossus muscle in rats in vivo. AB - The genioglossus (GG) muscle of the tongue, innervated by the hypoglossal motor nucleus (HMN), helps maintain an open airway for effective breathing. In vitro studies in neonatal rodents have separately characterized muscarinic and nicotinic receptor influences at the HMN but the net effects of combined nicotinic and muscarinic receptor activation and increased endogenous acetylcholine have not been determined in adult animals in vivo. Urethane anaesthetized, tracheotomized and vagotomised rats were studied. Microdialysis perfusion of acetylcholine into the HMN significantly decreased respiratory related GG activity (28.5 +/- 11.0% at a threshold dose of 0.1 mm). Application of the cholinergic agonists carbachol and muscarine have similar suppression effects (GG activity was decreased 11.8 +/- 4.3 and 20.5 +/- 5.8%, respectively, at 0.01 microm). Eserine, an acetylcholinesterase inhibitor, also decreased the amplitude of respiratory-related GG activity (36.4 +/- 11.3% at 1.0 microm) indicating that endogenous acetylcholine modulates GG activity. Although these results showed that suppression of GG activity predominates during cholinergic stimulation at the HMN, application of the nicotinic receptor agonist dimethyl-4 phenylpiperazinium iodide significantly increased tonic and respiratory-related GG activity (156 +/- 33% for respiratory activity at 1.0 mm) showing that excitatory responses are also present. Consistent with this, 100 microm carbachol decreased GG activity by 44.2 +/- 7.5% of control, with atropine (10 microm) reducing this suppression to 13.8 +/- 4.0% (P < 0.001). However, the nicotinic receptor antagonist dihydro-beta-erythroidine (100 microm) increased the carbachol-mediated suppression to 69.5 +/- 5.9% (P = 0.011), consistent with a role for nicotinic receptors in limiting the overall suppression of GG activity during cholinergic stimulation. Application of eserine to increase endogenous acetylcholine also showed that inhibitory muscarinic and excitatory nicotinic receptors together determine the net level of GG activity during cholinergic stimulation at the HMN. The results suggest that acetylcholine has mixed effects at the HMN with muscarinic-mediated GG suppression masking nicotinic excitation. PMID- 15817636 TI - Functional differentiation and organization of feline midlumbar commissural interneurones. AB - Interneurones interconnecting the two sides of the spinal cord (commissural interneurones) are critically important for interlimb coordination, but little is known about their organization. We have examined the inputs to commissural interneurones located in the midlumbar segments with projections to contralateral motor nuclei, aiming to determine whether they form distinct subpopulations. Based on intracellular records from 78 interneurones, two major non-overlapping subpopulations were identified: one monosynaptically excited by group II muscle afferents (n=10), the other monosynaptically excited by reticulospinal neurones (n=52). Monosynaptic input from group I muscle afferents and/or from vestibulospinal tract neurones was found in those with monosynaptic reticulospinal, but not group II input, and in a few other neurones (n=6). Only disynaptic input from these sources was found in the remaining 10 interneurones. Disynaptic excitatory input from ipsilateral and contralateral muscle afferents and from descending tracts was distributed less selectively and might mediate coexcitation of interneurones with monosynaptic afferent or descending input. The dominant disynaptic and polysynaptic input was, however, inhibitory. IPSPs were evoked from the descending tracts in a high proportion of the commissural interneurones that were monosynaptically excited by group II afferents (55%) and from group II afferents in a high proportion of the commissural interneurones that were monosynaptically excited by reticulospinal fibres (78%). This distribution suggests that the two subpopulations are activated differentially, rather than being coactivated, in either centrally initiated movements or reflex adjustments. This would be consistent with the previous demonstration that noradrenaline differentially affects commissural neurones of the two subpopulations. PMID- 15817637 TI - Elevated soluble ICAM-1 levels induce immune deficiency and increase adiposity in mice. AB - Elevated soluble intercellular adhesion molecule-1 (sICAM-1) levels have been found in many pathological conditions, including obesity. To determine the effects of elevated sICAM-1 on immune responses and metabolism, we generated a transgenic mouse model overexpressing the extracellular domain of mouse ICAM-1 in the liver. The mice, showing 10-fold higher sICAM-1 levels than wild-type mice, presented elevated neutrophil count. Despite this, after intraperitoneal injection of thioglycollate, neutrophil recruitment into the peritoneal cavity was reduced, and the delayed macrophage recruitment was also affected in the transgenic mice compared with wild-type mice. Inhibition of contact hypersensitivity response in the sICAM-1 transgenic mice was comparable to ICAM-1 deficient mice and characterized by significantly less ear swelling and inflammatory cell infiltration than in wild-type mice. sICAM-1transgenic mice were more susceptible to weight gain on a Western-type diet than wild-type mice, and older animals showed excessive fat accumulation, again reminiscent of ICAM-1 deficient mice. Together, these data indicate that sICAM-1 interferes with ICAM-1 mediated cell-cell interactions, which could produce immune-suppressant effects and alteration of metabolism in persons with high levels of this soluble adhesion receptor. PMID- 15817638 TI - Beta2-adrenoceptor regulation of the K+ channel iKCa1 in human mast cells. AB - Human mast cells express the intermediate conductance Ca2+-activated K+ channel iKCa1, which opens following IgE-dependent activation. This results in cell membrane hyperpolarization and potentiation of both Ca2+ influx and degranulation. Mast cell activation is attenuated following exposure to beta2 adrenoceptor agonists such as salbutamol, an effect postulated to operate via intracellular cyclic AMP. In this study, we show that salbutamol closes iKCa1 in mast cells derived from human lung and peripheral blood. Salbutamol (1-10 microM) inhibited iKCa1 currents following activation with both anti-IgE and the iKCa1 opener 1-EBIO, and was reversed by removing salbutamol or by the addition of the selective beta2-adrenoceptor antagonist and inverse agonist ICI 118551. Interestingly, ICI 118551 consistently opened iKCa1 in quiescent cells, suggesting that constitutive beta2-receptor signaling suppresses channel activity. Manipulation of intracellular cAMP, Galphai, and Galphas demonstrates that the beta2-adrenergic effects are consistent with a membrane-delimited mechanism involving Galphas. This is the first demonstration that gating of the iKCa1 channel is regulated by a G protein-coupled receptor and provides a clearly defined mechanism for the mast cell "stabilizing" effect of beta2-agonists. Furthermore, the degree of constitutive beta2-receptor "tone" may control the threshold for human mast cell activation through the regulation of iKCa1. PMID- 15817639 TI - C1-TEN is a negative regulator of the Akt/PKB signal transduction pathway and inhibits cell survival, proliferation, and migration. AB - We have previously identified C1 domain-containing phosphatase and TENsin homologue (C1-TEN) as being an intracellular binding partner for Axl receptor tyrosine kinase (RTK). C1-TEN is a tensin-related protein that houses an N terminal region with predicted structural similarity to PTEN. Here, we report our observations on the effects of ectopic expression of C1-TEN in HEK293 cells, which resulted in profound molecular and phenotypic changes. Stable expression of C1-TEN altered cellular morphology, with less cell spreading and weaker filamentous actin staining. Cells overexpressing C1-TEN were inhibited greatly in their proliferation and migration rates as compared with mock-transfected cells. Furthermore, serum starvation-induced apoptosis caused a twofold increase in caspase 3 activity in C1-TEN-overexpressing cells vs. mock cells. In addition, C1 TEN-overexpressing cells showed a markedly reduced phosphorylation of Akt/PKB kinase and its substrate GSK3, as well as reduced Akt enzymatic activity. No such effects on JNK were observed. Also, serum-stimulated activation of Akt was delayed in C1-TEN-overexpressing cells, while no difference in profile of ERK activation was observed. Furthermore, cells expressing a C1-TEN mutant where the putative phosphatase active site cysteine at position 231 was substituted for a serine displayed full restoration of both cell proliferation and Akt activation. In conclusion, C1-TEN appears to be a novel intracellular phosphatase that negatively regulates the Akt/PKB signaling cascade, and is similar to its relative PTEN in this respect. However, the particular domain organization of C1 TEN may enable it to regulate RTK and other signaling complexes that are linked to Akt/PKB signaling in a unique manner. PMID- 15817640 TI - Oxygen supply and nitric oxide scavenging by myoglobin contribute to exercise endurance and cardiac function. AB - Recent studies of myoglobin (Mb) knockout (myo-/-) mice have extended our understanding of Mb's diverse functions and have demonstrated a complex array of compensatory mechanisms. The present study was aimed at detailed analysis of cardiac function and exercise endurance in myo-/- mice and at providing evidence for Mb's functional relevance. Myo-/- isolated working hearts display decreased contractility (dP/dtmax 3883+/-351 vs. 4618+/-268 mmHg/sec, myo-/- vs. WT, P<0.005). Due to a shift in sympathetic/parasympathetic tone, heart rate is reduced in conscious myo mice-/- (615+/-33 vs. 645+/-27 bpm, myo-/- vs. WT, P<0.001). Oxygen consumption (VO2) under resting conditions (3082+/-413 vs. 4452+/-552 ml x kg(-1) x h(-1), myo-/- vs. WT, P<0.001) and exercise endurance, as determined by spiroergometry, are decreased (466+/-113 vs. 585+/-153 m, myo-/- vs. WT, P<0.01). Conscious myo-/- mice evaluated by echocardiography display lowered cardiac output (0.64+/-0.06 vs. 0.75+/-0.09 ml x min(-1) x g(-1), myo-/- vs. WT, P<0.001), impaired systolic shortening (60+/-3.5 vs. 65+/-4%, myo-/- vs. WT, P<0.001) and fail to respond to beta1-stimulation. Strikingly, the latter cardiac effects of Mb deficiency can be partially attenuated by NOS inhibition. Loss of Mb results in a distinct phenotype, even under resting conditions, and the importance of oxygen supply and nitric oxide scavenging by Mb is clearly demonstrated at the conscious animal level. PMID- 15817641 TI - Eczema drugs tacrolimus (Protopic) and pimecrolimus (Elidel): cancer concerns. PMID- 15817643 TI - Topographic maps of visual spatial attention in human parietal cortex. AB - Functional magnetic resonance imaging (fMRI) was used to measure activity in human parietal cortex during performance of a visual detection task in which the focus of attention systematically traversed the visual field. Critically, the stimuli were identical on all trials (except for slight contrast changes in a fully randomized selection of the target locations) whereas only the cued location varied. Traveling waves of activity were observed in posterior parietal cortex consistent with shifts in covert attention in the absence of eye movements. The temporal phase of the fMRI signal in each voxel indicated the corresponding visual field location. Visualization of the distribution of temporal phases on a flattened representation of parietal cortex revealed at least two distinct topographically organized cortical areas within the intraparietal sulcus (IPS), each representing the contralateral visual field. Two cortical areas were proposed based on this topographic organization, which we refer to as IPS1 and IPS2 to indicate their locations within the IPS. This nomenclature is neutral with respect to possible homologies with well-established cortical areas in the monkey brain. The two proposed cortical areas exhibited relatively little response to passive visual stimulation in comparison with early visual areas. These results provide evidence for multiple topographic maps in human parietal cortex. PMID- 15817644 TI - Topographic organization for delayed saccades in human posterior parietal cortex. AB - Posterior parietal cortex (PPC) is thought to play a critical role in decision making, sensory attention, motor intention, and/or working memory. Research on the PPC in non-human primates has focused on the lateral intraparietal area (LIP) in the intraparietal sulcus (IPS). Neurons in LIP respond after the onset of visual targets, just before saccades to those targets, and during the delay period in between. To study the function of posterior parietal cortex in humans, it will be crucial to have a routine and reliable method for localizing specific parietal areas in individual subjects. Here, we show that human PPC contains at least two topographically organized regions, which are candidates for the human homologue of LIP. We mapped the topographic organization of human PPC for delayed (memory guided) saccades using fMRI. Subjects were instructed to fixate centrally while a peripheral target was briefly presented. After a further 3-s delay, subjects made a saccade to the remembered target location followed by a saccade back to fixation and a 1-s inter-trial interval. Targets appeared at successive locations "around the clock" (same eccentricity, approximately 30 degrees angular steps), to produce a traveling wave of activity in areas that are topographically organized. PPC exhibited topographic organization for delayed saccades. We defined two areas in each hemisphere that contained topographic maps of the contra-lateral visual field. These two areas were immediately rostral to V7 as defined by standard retinotopic mapping. The two areas were separated from each other and from V7 by reversals in visual field orientation. However, we leave open the possibility that these two areas will be further subdivided in future studies. Our results demonstrate that topographic maps tile the cortex continuously from V1 well into PPC. PMID- 15817645 TI - Stimulation of melatonin receptors decreases calcium levels in xenopus tectal cells by activating GABA(C) receptors. AB - To investigate the physiological effects of melatonin receptors in the Xenopus tectum, we have used the fluorescent indicator Fluo-4 AM to monitor calcium dynamics of cells in tectal slices. Bath application of KCl elicited fluorescence increases that were reduced by melatonin. This effect was stronger at the end of the light period than at the end of the dark period. Melatonin increased gamma aminobutyric acid-C (GABA(C))-receptor activity, as demonstrated by the ability of the GABA(C)-receptor antagonists, picrotoxin and TPMPA, to abolish the effects of melatonin. In contrast, neither the GABA(A)-receptor antagonist bicuculline nor the GABA(B)-receptor antagonist CGP 35348 diminished the effects of melatonin. RT-PCR analyses revealed expression of the 3 known melatonin receptors, MT1 (Mel1(a)), MT2 (Mel1(b)), and Mel1(c). Because the effect of melatonin on tectal calcium increases was antagonized by an MT2-selective antagonist, 4-P-PDOT, we performed Western blot analyses with an antibody to the MT2 receptor; the data indicate that the MT2 receptor is expressed primarily as a dimeric complex and is glycosylated. The receptor is present in higher amounts at the end of the light period than at the end of the dark period, in a pattern complementary to the changes in melatonin levels, which are higher during the night than during the day. These results imply that melatonin, acting by MT2 receptors, modulates GABA(C) receptor activity in the optic tectum and that this effect is influenced by the light-dark cycle. PMID- 15817646 TI - Cyclic AMP cascade mediates the inhibitory odor response of isolated toad olfactory receptor neurons. AB - Odor stimulation may excite or inhibit olfactory receptor neurons (ORNs). It is well established that the excitatory response involves a cyclic AMP (cAMP) transduction mechanism that activates a nonselective cationic cyclic nucleotide gated (CNG) conductance, accompanied by the activation of a Ca2+-dependent Cl(-) conductance, both causing a depolarizing receptor potential. In contrast, odor inhibition is attributed to a hyperpolarizing receptor potential. It has been proposed that a Ca2+-dependent K+ (K(Ca)) conductance plays a key role in odor inhibition, both in toad and rat isolated olfactory neurons. The mechanism underlying odor inhibition has remained elusive. We assessed its study using various pharmacological agents and caged compounds for cAMP, Ca2+, and inositol 1,4,5-triphosphate (InsP3) on isolated toad ORNs. The odor-triggered K(Ca) current was reduced on exposing the cell either to the CNG channel blocker LY83583 (20 microM) or to the adenylyl cyclase inhibitor SQ22536 (100 microM). Photorelease of caged Ca2+ activated a Cl- current sensitive to niflumic acid (10 microM) and a K+ current blockable by charybdotoxin (20 nM) and iberiotoxin (20 nM). In contrast, photoreleased Ca2+ had no effect on cells missing their cilia, indicating that these conductances are confined to the cilia. Photorelease of cAMP induced a charybdotoxin-sensitive K+ current in intact ORNs. Photorelease of InsP3 did not increase the membrane conductance of olfactory neurons, arguing against a direct role of InsP3 in chemotransduction. We conclude that a cAMP cascade mediates the activation of the ciliary Ca2+-dependent K+ current and that the Ca2+ ions that activate the inhibitory current enter the cilia through CNG channels. PMID- 15817647 TI - Processing of modulated sounds in the zebra finch auditory midbrain: responses to noise, frequency sweeps, and sinusoidal amplitude modulations. AB - The avian auditory midbrain nucleus, the mesencephalicus lateralis, dorsalis (MLd), is the first auditory processing stage in which multiple parallel inputs converge, and it provides the input to the auditory thalamus. We studied the responses of single MLd neurons to four types of modulated sounds: 1) white noise; 2) band-limited noise; 3) frequency modulated (FM) sweeps, and 4) sinusoidally amplitude-modulated tones (SAM) in adult male zebra finches. Responses were compared with the responses of the same neurons to pure tones in terms of temporal response patterns, thresholds, characteristic frequencies, frequency tuning bandwidths, tuning sharpness, and spike rate/intensity relationships. Most neurons responded well to noise. More than one-half of the neurons responded selectively to particular portions of the noise, suggesting that, unlike forebrain neurons, many MLd neurons can encode specific acoustic components of highly modulated sounds such as noise. Selectivity for FM sweep direction was found in only 13% of cells that responded to sweeps. Those cells also showed asymmetric tuning curves, suggesting that asymmetric inhibition plays a role in FM directional selectivity. Responses to SAM showed that MLd neurons code temporal modulation rates using both spike rate and synchronization. Nearly all cells showed low-pass or band-pass filtering properties for SAM. Best modulation frequencies matched the temporal modulations in zebra finch song. Results suggest that auditory midbrain neurons are well suited for encoding a wide range of complex sounds with a high degree of temporal accuracy rather than selectively responding to only some sounds. PMID- 15817648 TI - Sensitivity of auditory cortical neurons to the locations of leading and lagging sounds. AB - We recorded unit activity in the auditory cortex (fields A1, A2, and PAF) of anesthetized cats while presenting paired clicks with variable locations and interstimulus delays (ISDs). In human listeners, such sounds elicit the precedence effect, in which localization of the lagging sound is impaired at ISDs less, similar10 ms. In the present study, neurons typically responded to the leading stimulus with a brief burst of spikes, followed by suppression lasting 100-200 ms. At an ISD of 20 ms, at which listeners report a distinct lagging sound, only 12% of units showed discrete lagging responses. Long-lasting suppression was found in all sampled cortical fields, for all leading and lagging locations, and at all sound levels. Recordings from awake cats confirmed this long-lasting suppression in the absence of anesthesia, although recovery from suppression was faster in the awake state. Despite the lack of discrete lagging responses at delays of 1-20 ms, the spike patterns of 40% of units varied systematically with ISD, suggesting that many neurons represent lagging sounds implicitly in their temporal firing patterns rather than explicitly in discrete responses. We estimated the amount of location-related information transmitted by spike patterns at delays of 1-16 ms under conditions in which we varied only the leading location or only the lagging location. Consistent with human psychophysical results, transmission of information about the leading location was high at all ISDs. Unlike listeners, however, transmission of information about the lagging location remained low, even at ISDs of 12-16 ms. PMID- 15817649 TI - Internal model of gravity for hand interception: parametric adaptation to zero gravity visual targets on Earth. AB - Internal model is a neural mechanism that mimics the dynamics of an object for sensory motor or cognitive functions. Recent research focuses on the issue of whether multiple internal models are learned and switched to cope with a variety of conditions, or single general models are adapted by tuning the parameters. Here we addressed this issue by investigating how the manual interception of a moving target changes with changes of the visual environment. In our paradigm, a virtual target moves vertically downward on a screen with different laws of motion. Subjects are asked to punch a hidden ball that arrives in synchrony with the visual target. By using several different protocols, we systematically found that subjects do not develop a new internal model appropriate for constant speed targets, but they use the default gravity model and reduce the central processing time. The results imply that adaptation to zero-gravity targets involves a compression of temporal processing through the cortical and subcortical regions interconnected with the vestibular cortex, which has previously been shown to be the site of storage of the internal model of gravity. PMID- 15817650 TI - Powerful, onset inhibition in the ventral nucleus of the lateral lemniscus. AB - The function of the ventral nucleus of the lateral lemniscus (VNLL), a secondary processing site within the auditory brain stem, is unclear. It is known to be a major source of inhibition to the inferior colliculus (IC). It is also thought to play a role in coding the temporal aspects of sound, such as onsets and the periodic components of complex stimuli. In vivo intracellular recordings from VNLL neurons (n = 56) in urethane anesthetized rats revealed the presence of large-amplitude, short-duration, onset inhibition in a subset of neurons (14.3%). This inhibition occurred before the first action potential (AP) elicited by noise or tone bursts, was broadly tuned to tonal frequency and was shown to delay the first AP. Our data suggest it is a result of an intrinsic circuit activated by the octopus cell pathway originating in the contralateral cochlear nucleus; this pathway is known to convey exquisitely timed and broadly tuned onset information. This powerful inhibition within the VNLL appears to control the timing of this structure's inhibitory output to higher centers, which has important auditory processing outcomes. The circuit also provides a pathway for fast, broadly tuned, onset inhibition to the IC. PMID- 15817651 TI - Peptidergic contribution to posttetanic potentiation at a central synapse of aplysia. AB - Posttetanic potentiation (PTP)-like phenomena appear to be mediated by a variety of mechanisms. Although neuropeptides are located in a large number of neurons and many neuropeptides, like PTP, can enhance synaptic transmission, there is a paucity of studies indicating that peptides may actually participate in PTP. Here, we utilize a single central synapse in the feeding circuit of Aplysia to investigate a possible peptidergic contribution to PTP in the CNS. The cholinergic command-like interneuron, cerebral-buccal interneuron 2 (CBI-2), contains two neuropeptides, feeding circuit activating peptide (FCAP) and cerebral peptide 2 (CP2). Previous studies showed that tetanic prestimulation or repeated stimulation of CBI-2, as well as perfusion of FCAP and CP2, increase the size of the cholinergic excitatory postsynaptic potentials (EPSPs) that CBI-2 evokes in the motoneurons B61/62 and shorten the latency to initiate B61/62 firing in response to CBI-2 stimulation. We used temperature-dependent suppression of peptide release and occlusion experiments to examine the possible contribution of FCAP and CP2 to PTP at the CBI-2 to B61/62 synapse. When peptide release was suppressed, perfusion of exogenous peptides increased the size of posttetanic EPSPs. In contrast, when peptide release was not suppressed, exogenous peptides did not enhance the size of posttetanic EPSPs, thus indicating occlusion. Temperature manipulation and occlusion experiments also indicated that peptides extend PTP duration. This peptide-dependent prolongation of PTP has functional consequences in that it extends the duration of time during which the latency to initiate B61/62 firing in response to CBI-2 stimulation is shortened. PMID- 15817652 TI - Neuronal activity dependent on anticipated and elapsed delay in macaque prefrontal cortex, frontal and supplementary eye fields, and premotor cortex. AB - In macaque monkeys performing a memory-guided saccade task for a reward of variable size, neuronal activity in several areas of frontal cortex is stronger when the monkey anticipates a larger reward. This effect might depend on either the size or the value of the reward. To distinguish between these possibilities, we recorded from neurons in frontal cortex while controlling value through a manipulation of time rather than amount. A cue presented at the beginning of each trial, predicted the length of the delay during which the monkey would have to maintain fixation before performing a saccade and receiving a reward of fixed size. Predicting a short delay had effects closely similar to those of predicting a large reward: 1) monkeys were more motivated when working for a reward at short delay, 2) neurons tended to fire more strongly before a short delay, 3) individual neurons firing more strongly before a short delay tended also to fire more strongly before a large reward, and 4) the tendency to fire more strongly before a short delay was far more pronounced in premotor areas caudal to the arcuate sulcus than in association areas rostral to it. The association areas, in contrast, were marked by a tendency for neurons to fire more strongly at the end of the long delay. We conclude that predicting a short delay, like predicting a large reward, induces an enhancement of neuronal activity related to motivational modulation of the monkey's preparatory state. PMID- 15817653 TI - p38 Mitogen-activated protein kinase (MAPK) is a key mediator in glucocorticoid induced apoptosis of lymphoid cells: correlation between p38 MAPK activation and site-specific phosphorylation of the human glucocorticoid receptor at serine 211. AB - Glucocorticoids (GCs) induce apoptosis in lymphoid cells through activation of the GC receptor (GR). We have evaluated the role of p38, a MAPK, in lymphoid cell apoptosis upon treatment with the synthetic GCs dexamethasone (Dex) or deacylcortivazol (DAC). The highly conserved phosphoprotein p38 MAPK is activated by specific phosphorylation of its threonine180 and tyrosine182 residues. We show that Dex and DAC stimulate p38 MAPK phosphorylation and increase the mRNA of MAPK kinase 3, a specific immediate upstream activator of p38 MAPK. Enzymatic assays confirmed elevated activity of p38 MAPK. Pharmacological inhibition of p38 MAPK activity was protective against GC-driven apoptosis in human and mouse lymphoid cells. In contrast, inhibition of the MAPKs, ERK and cJun N-terminal kinase, enhanced apoptosis. Activated p38 MAPK phosphorylates specific downstream targets. Because phosphorylation of the GR is affected by MAPKs, we examined its phosphorylation state in our system. We found serine 211 of the human GR to be a substrate for p38 MAPK both in vitro and intracellularly. Mutation of this site to alanine greatly diminished GR-driven gene transcription and apoptosis. Our results clearly demonstrate a role for p38 MAPK signaling in the pathway of GC induced apoptosis of lymphoid cells. PMID- 15817656 TI - Does increasing the grades of the knee osteoarthritis line drawing atlas alter its clinimetric properties? AB - OBJECTIVES: To (a) develop further logically derived line drawing atlases (LDAs) for grading radiographic knee osteoarthritis (OA); and (b) determine which is superior using metrological criteria. METHODS: A series of LDAs (-3 to +3, -4 to +4, and -5 to +5) were produced by (a) incorporating additional grades for osteophyte and joint space width (JSW) above the 0-3 pilot LDA, over an equivalent range of disease; and (b) adding negative grades for JSW. 121 sets of bilateral knee radiographs (standing, anteroposterior plus flexed skyline), plus serial views of 68 tibiofemoral joints (TFJs) and 36 patellofemoral joints were scored twice by one observer for each LDA. Minimum JSW of 50 radiograph sets was directly measured and awarded a categorical grade dependent upon the boundaries of each LDA grade. Time taken to grade 30 randomly selected knee radiograph sets was measured. RESULTS: Intraobserver reproducibility was similar for all LDAs, (weighted kappa: JSW = 0.85-0.87; osteophyte = 0.77-0.79), with no deterioration with increasing grades. Criterion validity favoured the -5 to +5 LDA, which was also quickest to use. All atlases showed similar responsiveness (standardised response mean: medial TFJ JSW = 0.78-0.83; medial femoral osteophyte = 0.61 0.73), with most sites compromised by small sample size, little change in score, and high variation between subjects. CONCLUSIONS: A set of LDAs was created illustrating the full range of normality/abnormality likely to be encountered in a community study of knee pain or OA. Despite superior validity and equivalent reproducibility, improved responsiveness of the -5 to +5 LDA was not confirmed. PMID- 15817655 TI - Antiphospholipid antibody tests: spreading the net. AB - OBJECTIVE: To examine the hypothesis that testing for new antiphospholipid antibody specificities may help to identify the antiphospholipid syndrome (APS) in patients with systemic lupus erythematosus (SLE) with thrombosis who are repeatedly negative for anticardiolipin antibodies (aCL) and/or lupus anticoagulant (LA). METHODS: Three groups of patients with SLE were studied: (a) SLE/APS (n = 56): 51 female, mean (SD) age 46 (11) years, fulfilling 1999 Sapporo criteria for the APS; (b) SLE/thrombosis (n = 56): 53 female, age 42.6 (12) years, all with a history of thrombosis and persistently negative for aCL and/or LA; (c) SLE only (n = 56): 53 female, age 40 (11) years, without a history of thrombotic events. aCL and LA were retested in all samples. All patients were tested for anti-beta(2)-glycoprotein I (anti-beta(2)GPI) and antiprothrombin antibodies (aPT) by coating prothrombin on irradiated plates or using phosphatidylserine-prothrombin complex as the antigen (aPS-PT). RESULTS: Anti beta(2)GPI were only present in patients from the SLE/APS group, all of whom were also positive for aCL. aPT and aPS-PT were also more commonly found in SLE/APS than in SLE/thrombosis or SLE only groups (54% v 5%, p<0.0001 or v 16%, p<0.0001 for aPT and 63% v 2%, p<0.0001 or v 11%, p<0.0001 for aPS-PT, respectively). No differences were found between SLE/thrombosis and SLE only groups (p = 1.5 for beta(2)GPI, p = 0.1 for aPT, and p = 0.1 for aPS-PT). CONCLUSION: Testing for aPT in patients with SLE with thrombosis, but persistently negative for aCL and LA, may be helpful in some selected cases. Anti-beta(2)GPI are not present in patients who are negative for aCL. PMID- 15817654 TI - Silencing of Fshr occurs through a conserved, hypersensitive site in the first intron. AB - Expression of the FSH receptor (Fshr) is restricted to testicular Sertoli cells and ovarian granulosa cells, thereby limiting the direct targets of FSH action to these somatic cells of the gonads. Earlier studies indicate that transcription of Fshr in the gonads requires elements outside the gene's immediate 5' flanking sequence. To help uncover candidate regulatory sequences, comparative genomics and deoxyribonuclease I hypersensitivity mapping were employed. A total of 156 evolutionarily conserved sequences were found, and partial deoxyribonuclease I hypersensitivity mapping across 45 kb of 5' flanking sequence and the first intron identified four hypersensitive sites, DHS1-4. Notably, DHS1 and DHS2 localized to conserved sites in the promoter region and exon 1 and correlated with the active state of the gene. DHS3 also corresponded to a conserved site (site 7) but was more pronounced in nonexpressing myoid cells, suggesting a role in gene silencing. Transient transfection analysis of DHS3 confirmed its role in gene silencing, a function that was promoter, cell type, and position dependent. Protein-DNA binding studies on DHS3 revealed that octamer transcription factor 1 (OCT-1) and GATA-4 bound site 7, in vitro, and transient transfection analysis showed that their binding sites were required for silencing activity. Furthermore, chromatin immunoprecipitation revealed that OCT-1 bound to site 7 in the endogenous gene, but only in myoid cells. In contrast, GATA-1 bound site 7 predominantly in Sertoli cells, suggesting that it attenuates silencer activity. The findings reveal that OCT-1 binds within DHS3 to silence Fshr transcription and implicate members of the GATA family in the modulation of this activity. PMID- 15817657 TI - Involvement of neurotrophins and their receptors in spondyloarthritis synovitis: relation to inflammation and response to treatment. AB - OBJECTIVE: To investigate whether expression of the four members of the neurotrophin (NT) family and their four corresponding receptors is related to synovial inflammation in patients with spondyloarthritis (SpA). MATERIAL AND METHODS: Synovial fluid (SF) and serum NTs and their receptors were measured by ELISA. Immunohistochemistry was used for synovial tissue biopsy specimens from patients with SpA, rheumatoid arthritis, and osteoarthritis (OA). In SpA synovium, immunoreactivity of the receptors trkA and NGFRp75 was also assessed before and after 12 weeks of treatment with the monoclonal anti-tumour necrosis factor alpha antibody, infliximab. RESULTS: mRNA transcripts of all NTs and receptors were expressed in the inflamed synovium. At the protein level, brain derived neurotrophic factor and NT-3 were significantly higher in the SF of patients with SpA than in those with OA. In contrast, ELISA of serum samples showed that the highest member in SpA was NT-4. Immunohistochemistry demonstrated that the NT receptors trkA and NGFRp75 were highly expressed in the inflamed synovium of patients with SpA, correlating with vascularity and lymphoid aggregates, respectively. Additionally, immunoreactivity of both receptors was significantly decreased after infliximab treatment. CONCLUSIONS: NTs and their receptors are expressed in inflamed peripheral joints of patients with SpA. Their expression is not constitutive but related to inflammation and they may be involved in the local disease processes. PMID- 15817659 TI - Salivary gland and peripheral blood T helper 1 and 2 cell activity in Sjogren's syndrome compared with non-Sjogren's sicca syndrome. AB - OBJECTIVES: To investigate whether differences in T helper (Th) 1 and Th2 cell activity in salivary glands ("local") or ("peripheral") blood can discriminate between Sjogren's syndrome (SS) and non-Sjogren's sicca syndrome (nSS-sicca). Additionally, to study relationships of local and peripheral Th cell activities with each other and with disease activity measures. METHODS: 62 sicca patients (32 with SS, 30 with nSS-sicca) were studied. Local Th1 (interferon gamma (IFNgamma)) and Th2 (interleukin (IL) 4) activity were determined using immunohistochemistry. T cell production of IFNgamma and IL4 in peripheral blood (PB) was determined by ELISA. Erythrocyte sedimentation rate (ESR) and serum IgG were considered disease activity measures. RESULTS: ESR and serum IgG were higher in patients with SS than in patients with nSS-sicca. Local Th1 cell activity was higher and PB Th1 activity lower in patients with SS than in those with nSS sicca. Th2 cell activity did not differ significantly between the patient groups. The ratio IFNgamma/IL4 was higher in salivary glands and lower in PB in patients with SS than in patients with nSS-sicca. Local and peripheral Th1 and Th2 cell activities correlated with ESR and serum IgG levels. ESR, serum IgG, and local or peripheral Th1 or Th2 cell activity did not discriminate between patients with SS and nSS-sicca. CONCLUSIONS: An imbalance between Th1 and Th2 activity in sicca patients is clearly related to the severity of disease, but cannot be used to distinguish between patients with SS and those with nSS-sicca. PMID- 15817658 TI - Oestrogen receptor {alpha} gene polymorphisms in systemic lupus erythematosus. AB - OBJECTIVE: To analyse associations of two oestrogen receptor alpha (ORalpha) gene polymorphisms in 260 patients with SLE from northern Sweden. The two polymorphisms, PvuII T/C and the XbaI A/G, are located in the first intron of the ORalpha gene. METHODS: All patients fulfilling at least four of the ACR criteria for SLE were consecutively recruited during one year. The SLEDAI score and SLICC damage index were recorded. 670 individuals from the same geographical area served as controls. DNA from the patients and controls was extracted and genotyped using the 5' nuclease assay with an ABI PRISM 7900HT instrument. The genotype/phenotype relationships were calculated using SPSS. RESULTS: The unusual PvuII C allele was associated with malar rash and the unusual XbaI G allele with photosensitivity (p = 0.001, OR = 2.53, 95% CI = 1.43 to 4.47 and p = 0.007, OR = 2.12, 95% CI = 1.22 to 3.66, respectively). The common XbaI AA genotype was associated with serositis (p = 0.013, OR = 1.92, 95% CI = 1.15 to 3.22). Based on the SLICC damage index associations of the common TT genotype and AA genotype with cognitive impairment were identified (p = 0.018, OR = 2.47, 95% CI = 1.17 to 5.25 and p = 0.018, OR = 2.75, 95% CI = 1.19 to 6.38 respectively). There was also an association of the XbaI AA genotype with the angina/coronary artery bypass variable (p = 0.042, OR = 2.58, 95% CI = 1.03 to 6.43). Of the variables describing disease severity and duration it was found that carriers of the unusual PvuII C allele showed a later onset of SLE (p = 0.02) and carriers of the unusual XbaI G allele a lower SLICC damage index. CONCLUSIONS: The unusual PvuII C and XbaI G alleles were associated with a milder form of SLE characterised by skin manifestations, later onset, and less organ damage. PMID- 15817660 TI - Occurrence and risk factors for falls in rheumatoid arthritis. AB - OBJECTIVES: To determine the one year period prevalence of falls by age and sex in patients with rheumatoid arthritis and the influence of concurrent medical treatment and disability on the occurrence of falls in this group. METHODS: A consecutive series of rheumatoid patients aged 35 years and over, attending hospital outpatient clinics at Hope hospital, Salford, were asked to complete an interview assisted questionnaire which asked about the occurrence and number of falls in the previous 12 months. SUBJECTS: who took part were asked about current treatment with antihypertensive agents, diuretics, sedatives or hypnotics, antidepressants, and a history of previous hip/knee surgery. They also completed the health assessment questionnaire (HAQ). Logistic regression was used to determine the association between these variables and falls in the previous 12 months. RESULTS: 253 men and women, mean age 62 years, were studied, and 84 (33%) reported falling in the previous year (36% of women and 26% of men). Of these, 52% had fallen on more than one occasion. There was no important increase in the frequency of falls with age. After adjusting for age and sex, those who had fallen in the previous year were more likely to report taking antidepressant treatment (odds ratio (OR) = 2.09) and to have impairment in both walking (OR = 1.37) and rising (OR = 1.41). The HAQ score was higher in those who reported a fall than those who did not, though the difference was not statistically significant. CONCLUSIONS: In this hospital based survey, one in three patients with rheumatoid arthritis reported falling in the previous 12 months. Falls were associated with self reported impairment in lower limb function. PMID- 15817661 TI - Prevalence of spondyloarthropathies in France: 2001. AB - OBJECTIVE: To estimate the prevalence of spondyloarthropathies (SpAs) in France in a multiregional representative sample in the year 2001. METHODS: A two stage random sample was constituted in seven areas from the national telephone directory and the next birthday method in each household. Interviewers were patient-members of self help groups trained to administer telephone surveys using a validated questionnaire for detecting inflammatory joint disease. Quality of data collection was controlled periodically. SpA was confirmed by the patient's rheumatologist or by clinical examination. Prevalence estimates after probability sampling correction were standardised for age and sex (1999 national census). RESULTS: Among the 15 219 anonymous telephone numbers selected, 3.6% were places of work or secondary residences and were excluded. The phone interview participation rate ranged across regions from 55.1 to 69.9%. 3554 men and 5841 women were included in the study. Twenty nine cases of SpA were confirmed. All but one fulfilled ESSG criteria. Mean age was 47 years (range 21-78). The overall prevalence standardised for age and sex was 0.30% (95% confidence interval (CI) 0.17 to 0.46). Prevalence was similar in women (0.29% (95% CI 0.14 to 0.49)) and men (0.31 % (95% CI 0.12 to 0.60)). Geographical analysis by department clustering found no significant differences. The prevalence of SpA was as high as that of rheumatoid arthritis. CONCLUSION: Prevalence of SpA in France was 0.30% in 2001, with no difference between women and men. Ankylosing spondylitis and psoriatic arthritis were the most common SpA subsets. PMID- 15817662 TI - Complete inhibition of goiter in mice requires combined gene therapy modification of angiopoietin, vascular endothelial growth factor, and fibroblast growth factor signaling. AB - In goiter, increased expression of growth factors and their receptors occurs. We have inhibited the action of some of these growth factors, alone and in combination, to determine which are important in goitrogenesis. Recombinant adenovirus vectors (RAds) expressing truncated, secreted forms of human Tie2 (RAd sTie2) and vascular endothelial growth factor receptor 1 (RAd-sVEGFR1) or a truncated, dominant-negative fibroblast growth factor receptor 1 (RAdDN-FGFR1) were used. Goiters in mice were induced by feeding an iodide-deficient diet, containing methimazole and sodium perchlorate. RAds were administered to mice simultaneously with the goitrogenic regimen, which was continued for 14 d. RAd treatment did not significantly affect increases in TSH or reductions in thyroid hormone or thyroid hyperactivity seen in goitrogen-treated controls mice, suggesting no effect on pituitary or thyroid responses to hypothyroidism. In control goiters, a 4-fold increase in vascular volume accompanied a 2-fold increase in thyroid mass. Complete inhibition of these increases was found when animals were treated with the three RAds in combination. In thyroids from three RAd-treated animals, there was marked, significant inhibition of Tie2, FGFR1, VEGFR1, FGF-2, and VEGF expression, compared with control goiters. When used individually, RAdDN-FGFR1 partially prevented goiter and RAd-sVEGFR1 partially reduced vascular volume. Their effects were not additive. RAd-sTie2 did not reduce goiter mass or vascular volume when used alone but was essential for complete goiter inhibition. VEGF and VEGFR1 expression was reduced in these thyroids. Limitation of physiologic organ growth is complex, requiring inhibition of multiple, interdependent growth factor axes. PMID- 15817663 TI - Luteinizing hormone-induced extracellular-signal regulated kinase activation differently modulates progesterone and androstenedione production in bovine theca cells. AB - It has been reported that gonadotropins promoted phosphorylation of ERK/MAPK in granulosa cells. However, little is known about the effects of gonadotropin on ERK activity in theca cells. This study explores how LH/forskolin controls ERK phosphorylation in cultured bovine theca cells. Effects of ERK on steroidogenesis were also investigated. Phosphorylation of ERK in bovine theca cells was augmented by LH and forskolin in 5 min; it decreased thereafter below basal levels in 20 min. Nevertheless, phosphorylation of the ERK kinase, MEK, was unaffected. Addition of H89 (a protein kinase A inhibitor) significantly reduced the effect of LH/forskolin on ERK phosphorylation. A potent MEK inhibitor PD98059 eliminated ERK phosphorylation and augmented progesterone production concomitantly with the elevation of intracellular steroidogenic acute regulatory protein mRNA in LH/forskolin-stimulated theca cells. In contrast to progesterone production, androgen production was diminished significantly by inhibition of ERK with decreased intracellular P450c17 mRNA levels. Taking these results together, we conclude that LH/cAMP leads to phosphorylation of ERK in a biphasic manner through MEK-independent pathway in bovine theca cells. Protein kinase A-induced phosphatase could possibly contribute to the phosphorylation process. Furthermore, modulation of ERK phosphorylation involves control of thecal steroidogenesis via modulation of the expression of steroidogenic acute regulatory protein and P450c17. PMID- 15817664 TI - Follicular cells acquire sertoli cell characteristics after oocyte loss. AB - Although it has been suggested that in mammals the loss of female germ cells may induce the masculinization of the ovarian compartment, there has been as yet no conclusive demonstration. To directly address that question, the present study has been designed to determine the fate of follicular cells after oocyte loss. Using gamma-irradiation to selectively deplete oocytes in nongrowing follicles in female rats, we show that follicular cells in oocyte-depleted follicles survive, proliferate, and subsequently acquire morphological characteristics of Sertoli cells: elongated cytoplasm, basal location of the nucleus, and specific Sertoli cell junctions, the ectoplasmic specializations. These Sertoli-like cells express, however, the female-specific marker FOXL2 (Forkhead L2) but not the male sex-specific marker SOX-9 (Sry-type high-mobility-group box transcription factor 9) underlying the maintenance of molecular characteristics of granulosa cells. Before transdifferentiating into Sertoli-like cells, follicular cells of oocyte depleted follicles initiate the expression of anti-Mullerian hormone and inhibin alpha-subunit that are typically synthesized by granulosa cells from the onset of follicular growth. Experimental modifications of the endocrine balance of the irradiated females show that there is a close relationship between plasma FSH levels and the occurrence of Sertoli-like cells. In addition to providing experimental evidence for the crucial role of the oocyte in granulosa cell phenotype maintenance, these results emphasize that the transdifferentiation of granulosa cells into Sertoli cells occurs in a multistep fashion, requiring the maturation of granulosa cells and depending on the endocrine milieu. PMID- 15817665 TI - Estrogen can act via estrogen receptor alpha and beta to protect hippocampal neurons against global ischemia-induced cell death. AB - Estradiol at physiological concentrations intervenes in apoptotic death cascades and ameliorates neuronal death in experimental models of focal and global ischemia. The cellular targets that mediate estradiol protection of hippocampal neurons in global ischemia are, however, unclear. The present study examined the hypothesis that estradiol protects hippocampal neurons in ovariectomized rats via estrogen receptor (ER)alpha and/or beta. Estradiol (14 d pretreatment) afforded robust protection of CA1 neurons against global ischemia-induced death. The broad spectrum ER antagonist ICI 182,780 (intracerebroventricularly, 0 and 12 h after ischemia) abolished estrogen protection, consistent with a role for ERs. To evaluate the potential roles of ERalpha vs. ERbeta in estrogen protection, we administered subtype-selective agonists for 14 d before and 7 d after ischemia. The ERalpha-selective agonist propyl pyrazole triol (PPT, 10 mg/kg) and ERbeta selective agonist WAY 200070-3 (1 mg/kg) produced nearly complete protection of CA1 neurons in approximately 50% of the animals. PPT, but not WAY 200070-3, at doses used for protection, elicited lordosis, induced negative feedback inhibition of LH release, and reduced weight gain. These findings establish the efficacy of the PPT dose in neuroendocrine assays and specificity of WAY 200070-3 for ERbeta. We also examined the ability of estradiol and neuronal injury to regulate ERalpha and ERbeta expression. Both estradiol and global ischemia markedly increased ERalpha, but not ERbeta, protein in CA1. These data indicate that estradiol can act via ERalpha and ERbeta to protect CA1 neurons from global ischemia-induced death and that both estradiol and global ischemia modulate ERalpha expression in hippocampal CA1. PMID- 15817666 TI - Increased pituitary vascular endothelial growth factor-a in dopaminergic D2 receptor knockout female mice. AB - Vascular endothelial growth factor (VEGF)-A is an important angiogenic cytokine in cancer and pathological angiogenesis and has been related to the antiangiogenic activity of dopamine in endothelial cells. We investigated VEGF expression, localization, and function in pituitary hyperplasia of dopamine D2 receptor (D2R)-knockout female mice. Pituitaries from knockout mice showed increased protein and mRNA VEGF-A expression when compared with wild-type mice. In wild-type mice, prolonged treatment with the D2R antagonist, haloperidol, enhanced pituitary VEGF expression and prolactin release, suggesting that dopamine inhibits pituitary VEGF expression. VEGF expression was also increased in pituitary cells from knockout mice, even though these cells proliferated less in vitro when compared with wild-type cells, as determined by 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium proliferation assay, proliferating cell nuclear antigen expression, and [(3)H]thymidine incorporation. In contrast to other animal models, estrogen did not increase pituitary VEGF protein and mRNA expression and lowered serum prolactin secretion in vivo and in vitro in both genotypes. VEGF (10 and 30 ng/ml) did not modify pituitary cell proliferation in either genotype and increased prolactin secretion in vitro in estrogen-pretreated cells of both genotypes. But conditioned media from D2R(-/-) cells enhanced human umbilical vein cell proliferation, and this effect could be partially inhibited by an anti VEGF antiserum. Finally, using dual-labeling immunofluorescence and confocal laser microscopy, we found that in the hyperplastic pituitaries, VEGF-A was mostly present in follicle-stellate cells. In conclusion, pituitary VEGF expression is under dopaminergic control, and even though VEGF does not promote pituitary cellular proliferation in vitro, it may be critical for pituitary angiogenesis through paracrine actions in the D2R knockout female mice. PMID- 15817667 TI - Differential expression of procollagen lysine 2-oxoglutarate 5-deoxygenase and matrix metalloproteinase isoforms in hypothyroid rat ovary and disintegration of extracellular matrix. AB - Hypothyroid-induced reproductive malfunction in both the sexes is a common phenomenon of global concern. In an attempt to characterize the differentially expressed genes that might be responsible for these disorders, we have identified a number of clones in hypothyroid rat ovary by subtractive hybridization. One such clone is procollagen lysyl hydroxylase2 (Plod-2), the key enzyme for the first step of collagen biosynthetic pathway, which was down-regulated in hypothyroid condition. We have also demonstrated the reduced expression of other isoforms of Plods, namely Plod-1 and -3 in hypothyroid rat ovary. The current studies are the first of their kind to report that thyroid hormone regulates the Plod gene in rat ovary. Moreover, we have shown the up-regulation of matrix degrading enzyme(s), matrix metalloproteinase(s) in the hypothyroid rat ovary, whereas the tissue-inhibitory metalloproteinase is down-regulated. Finally, the results of the present studies indicate that in hypothyroid condition, collagen biosynthesis in ovary seems to be disturbed with concomitant enhancement in collagen degradation, resulting in disintegration of overall ovarian structure. PMID- 15817668 TI - Differential regulation of sodium/iodide symporter gene expression by nuclear receptor ligands in MCF-7 breast cancer cells. AB - The sodium/iodide symporter (NIS) mediates iodide uptake in lactating breast tissue and is expressed in some breast cancers. We have previously demonstrated that all-trans retinoic acid (tRA) stimulates NIS gene expression and the selective cytotoxic effect of beta-emitting radioiodide-131 ((131)I) in both in vitro and in vivo MCF-7 breast cancer cell systems. We studied the ability of natural and synthetic retinoids, in combination with other nuclear receptor ligands, to achieve greater and more sustained induction of NIS in MCF-7 cells and enhance (131)I-mediated cytotoxicity. Selective stimulation of retinoic acid receptor (RAR) beta/gamma produced marked NIS induction; and selective stimulation of RARalpha, RARgamma, or retinoid X receptor produced more modest induction. Maximal NIS induction was seen with 9-cis retinoic acid and AGN190168, a RAR beta/gamma-agonist. Dexamethasone (Dex), but not the other nuclear receptor ligands, in combination with tRA synergistically induced iodide uptake and NIS mRNA expression, predominantly by prolonging NIS mRNA half-life. The addition of Dex reduced the EC(50) of tRA for NIS stimulation to approximately 7%, such that 10(-7) m tRA with addition of Dex enhanced iodide uptake and selective cytotoxicity of (131)I greater than 10(-6) m tRA alone. AGN190168 combined with Dex synergistically increased iodide uptake and significantly prolonged induction (5 d) of iodide uptake compared with that induced by the combination of tRA/Dex or 9-cis retinoic acid/Dex. The addition of Dex reduced the effective dose of retinoid and prolonged the induction of NIS, especially with AGN190168, suggesting higher efficacy of (131)I after combination treatment. PMID- 15817669 TI - Human growth hormone-releasing factor (hGRF)1-29-albumin bioconjugates activate the GRF receptor on the anterior pituitary in rats: identification of CJC-1295 as a long-lasting GRF analog. AB - In vivo bioconjugation to the free thiol on Cys34 of serum albumin by a strategically placed reactive group on a bioactive peptide is a useful tool to extend plasma half-life. Three maleimido derivates of human GH-releasing factor (hGRF)(1-29) were synthesized and bioconjugated to human serum albumin ex vivo. All three human serum albumin conjugates showed enhanced in vitro stability against dipeptidylpeptidase-IV and were bioactive in a GH secretion assay in cultured rat anterior pituitary cells. When the maleimido derivatives were individually administered sc to normal male Sprague Dawley rats, an acute secretion of GH was measured in plasma. The best compound, CJC-1295, showed a 4 fold increase in GH area under the curve over a 2-h period compared with hGRF(1 29). CJC-1295, a tetrasubstituted form of hGRF(1-29) with an added N epsilon-3 maleimidopropionamide derivative of lysine at the C terminus, was selected for further pharmacokinetic evaluation, where it was found to be present in plasma beyond 72 h. A Western blot analysis of the plasma of a rat injected with CJC 1295 showed the presence of a CJC-1295 immunoreactive species on the band corresponding to serum albumin, appearing after 15 min and remaining in circulation beyond 24 h. These results led to the identification of CJC-1295 as a stable and active hGRF(1-29) analog with an extended plasma half-life. PMID- 15817670 TI - Potential role for human cytochrome P450 3A4 in estradiol homeostasis. AB - Previously, a human CYP3A4-transgenic (Tg-CYP3A4) mouse line was reported to exhibit enhanced metabolism of midazolam by cytochrome P450 3A4 (CYP3A4) expressed in small intestine. Here we show that expression of CYP3A4 and murine cyp3a and cyp2b was both age and sex dependent. CYP3A4 was expressed in the livers of male and female Tg-CYP3A4 mice at 2 and 4 wk of age. Since 6 wk, CYP3A4 was undetectable in male livers, whereas it was constitutively expressed in female livers at decreased levels (3- to 5-fold). Pregnenolone 16alpha carbonitrile markedly induced hepatic CYP3A4 expression, and the level was higher in females than males. Induction of intrinsic murine cyp3a and cyp2b was also sex dependent. Tg-CYP3A4 females were found to be deficient in lactation, leading to a markedly lower pup survival. The mammary glands of the Tg-CYP3A4 lactating mothers had underdeveloped alveoli with low milk content. Furthermore, beta casein and whey acidic protein mRNAs were expressed at markedly lower levels in Tg-CYP3A4 pregnant and nursing mouse mammary glands compared with wild-type mice. This impaired lactation phenotype was associated with significantly reduced serum estradiol levels in Tg-CYP3A4 mice. A pharmacokinetic study revealed that the clearance of iv administrated [(3)H]estradiol was markedly enhanced in Tg-CYP3A4 mice compared with wild-type mice. These results suggest that CYP3A4 may play an important role in estradiol homeostasis. This may be of concern for treatment of pregnant and lactating women because CYP3A4 gene expression and enzymatic activity can be potentially modified by CYP3A4 inhibitors or inducers in medications, supplements, beverages, and diet. PMID- 15817671 TI - Antiobesity effect of a melanin-concentrating hormone 1 receptor antagonist in diet-induced obese mice. AB - Melanin-concentrating hormone (MCH) is a cyclic orexigenic peptide expressed in the lateral hypothalamus, which plays an important role in regulating energy balance. To elucidate the physiological role of MCH in obesity development, the present study examined the effect of a selective MCH1 receptor (MCH1R) antagonist in the diet-induced obesity mouse model. The MCH1R antagonist has high affinity and selectivity for MCH-1R and potently inhibits intracerebroventricularly injected MCH-induced food intake in Sprague Dawley rats. Chronic intracerebroventricular infusion of the MCH1R antagonist (7.5 microg/d) completely suppressed body weight gain in diet-induced obese mice during the treatment periods and significantly decreased cumulative food intake, by 14%. Carcass analysis showed that the MCH1R antagonist resulted in a selective decrease of body fat in the diet-induced obese mice. In addition, the MCH1R antagonist ameliorated the obesity-related hypercholesterolemia, hyperinsulinemia, hyperglycemia, and hyperleptinemia. These results indicate that MCH has a major role in the development of diet-induced obesity in mice and that a MCH1R antagonist might be a useful candidate as an antiobesity agent. PMID- 15817672 TI - Estrogen--the good, the bad, and the unexpected. PMID- 15817673 TI - Selective depletion of alloreactive donor lymphocytes: a novel method to reduce the severity of graft-versus-host disease in older patients undergoing matched sibling donor stem cell transplantation. AB - We have selectively depleted host-reactive donor T cells from peripheral blood stem cell (PBSC) transplant allografts ex vivo using an anti-CD25 immunotoxin. We report a clinical trial to decrease graft-versus-host disease (GVHD) in elderly patients receiving selectively depleted PBSC transplants from HLA-identical sibling donors. Sixteen patients (median age, 65 years [range, 51-73 years]), with advanced hematologic malignancies underwent transplantation following reduced-intensity conditioning with fludarabine and either cyclophosphamide (n = 5), melphalan (n = 5), or busulfan (n = 6). Cyclosporine was used as sole GVHD prophylaxis. The allograft contained a median of 4.5 x 10(6) CD34 cells/kg (range, 3.4-7.3 x 10(6) CD34 cells/kg) and 1.0 x 10(8)/kg (range, 0.2-1.5 x 10(8)/kg) selectively depleted T cells. Fifteen patients achieved sustained engraftment. The helper T-lymphocyte precursor (HTLp) frequency assay demonstrated successful (mean, 5-fold) depletion of host-reactive donor T cells, with conservation of third-party response in 9 of 11 cases tested. Actuarial rates of acute GVHD were 46% +/- 13% for grades II to IV and 12% +/- 8% for grades III to IV. These results suggest that allodepletion of donor cells ex vivo is clinically feasible in older patients and may reduce the rate of severe acute GVHD. Further studies with selectively depleted transplants to evaluate graft versus-leukemia (GVL) and survival are warranted. PMID- 15817674 TI - A clinically relevant SCID-hu in vivo model of human multiple myeloma. AB - We developed a novel in vivo multiple myeloma (MM) model by engrafting the interleukin 6 (IL-6)-dependent human MM cell line INA-6 into severe combined immunodeficiency (SCID) mice previously given implants of a human fetal bone chip (SCID-hu mice). INA-6 cells require either exogenous human IL-6 (huIL-6) or bone marrow stromal cells (BMSCs) to proliferate in vitro. In this model, we monitored the in vivo growth of INA-6 cells stably transduced with a green fluorescent protein (GFP) gene (INA-6GFP+ cells). INA-6 MM cells engrafted in SCID-hu mice but not in SCID mice that had not been given implants of human fetal bone. The level of soluble human IL-6 receptor (shuIL-6R) in murine serum and fluorescence imaging of host animals were sensitive indicators of tumor growth. Dexamethasone as well as experimental drugs, such as Atiprimod and B-B4-DM1, were used to confirm the utility of the model for evaluation of anti-MM agents. We report that this model is highly reproducible and allows for evaluation of investigational drugs targeting IL-6-dependent MM cells in the human bone marrow (huBM) milieu. PMID- 15817676 TI - Arf6: a new player in FcgammaRIIIA lymphocyte-mediated cytotoxicity. AB - The activation of phosphoinositide metabolism represents a critical step in the signaling pathways leading to the activation of cytolytic machinery, but its regulation is partially understood. We report here that the stimulation of the low-affinity receptor for immunoglobulin G (IgG) (FcgammaRIIIA, CD16) on primary human natural killer (NK) cells induces a phosphatidylinositol 3-kinase (PI3K) dependent activation of the small G protein Arf6. We first demonstrate a functional role for Arf6-dependent signals in the activation of the antibody dependent cellular cytotoxicity (ADCC) attributable to the control of secretion of lytic granule content. We also show that Arf6 couples CD16 to the lipid modifying enzymes phosphatidylinositol4phosphate 5-kinase type I alpha (PI5KIalpha) and phospholipase D (PLD) that are involved in the control of granule secretion; Arf6, but not Rho family small G proteins RhoA and Rac1, is required for receptor-induced PI5KIalpha membrane targeting as well as for PI5KIalpha and PLD activation. Our findings suggest that Arf6 plays a crucial role in the generation of a phosphatidylinositol4,5-bisphosphate (PIP2) plasma membrane pool required for cytolytic granule-mediated target cell killing. PMID- 15817675 TI - Tie2 activation contributes to hemangiogenic regeneration after myelosuppression. AB - Chemotherapy- or radiation-induced myelosuppression results in apoptosis of cycling hematopoietic cells and induces regression of bone marrow (BM) sinusoidal vessels. Moreover, timely regeneration of BM neovessels is essential for reconstitution of hematopoiesis. However, the identity of angiogenic factors that support reconstitution of BM's vasculature is unknown. Here, we demonstrate that angiopoietin/tyrosine kinase with immunoglobulin and epidermal growth factor homology domains-2 (Tie2) signaling contributes to the assembly and remodeling of BM neovessels after myelosuppression. Using transgenic mice where the Tie2 promoter drives the reporter LacZ gene (Tie2-LacZ), we demonstrate that at steady state, there was minimal expression of Tie2 in the BM vasculature. However, after 5-fluorouracil (5-FU) treatment, there was a rapid increase in plasma vascular endothelial growth factor A (VEGF-A) levels and expansion of Tie2-positive neovessels. Inhibition of Tie2 resulted in impaired neoangiogenesis, leading to a delay in hematopoietic recovery. Conversely, angiopoietin-1 (Ang-1) stimulated hematopoiesis both in wild-type and thrombopoietin-deficient mice. In addition, Ang-1 shortened the duration of chemotherapy-induced neutropenia in wild-type mice. Exogenous VEGF-A and Ang-1 stimulated Tie2 expression in the BM vasculature. These data suggest that VEGF-A-induced up-regulation of Tie2 expression on the regenerating vasculature after BM suppression supports the assembly of sinusoidal endothelial cells, thereby promoting reconstitution of hematopoiesis. Angiopoietins may be clinically useful to accelerate hemangiogenic recovery after myelosuppression. PMID- 15817677 TI - Distinctive gene expression pattern in VH3-21 utilizing B-cell chronic lymphocytic leukemia. AB - The usage of the immunoglobulin (Ig) V(H)3-21 gene is associated with poor prognosis in B-cell chronic lymphocytic leukemia (B-CLL) despite V(H) gene mutation status. Many V(H)3-21+ patients also display restricted heavy- and light chain Ig gene rearrangements, implying a role of antigen selection in disease development. To explore the specific phenotypic/genotypic features among V(H)3 21+ B-CLLs, we compared gene expression patterns in 15 V(H)3-21+ and 24 non-V(H)3 21 patients (11 with unmutated and 13 with mutated V(H) genes) using Affymetrix microarray analysis (approximately 12,500 genes). A distinct expression profile was identified for V(H)3-21+ patients in contrast to the Ig-unmutated and mutated groups. By applying different algorithms, the data enabled an efficient class discrimination of the V(H)3-21+ subset based on 27 or 57 genes. A set of genes was sorted out which, using different analytical methods, consistently gave a distinction between V(H)3-21+ and non-V(H)3-21 samples. Several of these genes are involved in regulation of DNA replication/cell-cycle control, transcription and protein kinase activity, which may render the V(H)3-21+ cells with a higher proliferative drive. However, no clear evidence of increased B-cell receptor signaling was found in the V(H)3-21+ group. Altogether, our identification of a specific V(H)3-21 profile may provide insights into the pathogenesis of the V(H)3 21+ subgroup. PMID- 15817678 TI - A crucial role for reactive oxygen species in RANKL-induced osteoclast differentiation. AB - Signaling by receptor activator of NF-kappaB (nuclear factor-kappaB) ligand (RANKL) is essential for differentiation of bone marrow monocyte-macrophage lineage (BMM) cells into osteoclasts. Here, we show RANKL stimulation of BMM cells transiently increased the intracellular level of reactive oxygen species (ROS) through a signaling cascade involving TNF (tumor necrosis factor) receptor associated factor (TRAF) 6, Rac1, and NADPH (nicotinamide adenine dinucleotide phosphate) oxidase (Nox) 1. A deficiency in TRAF6 or expression of a dominant interfering mutant of TRAF6 blocks RANKL-mediated ROS production. Application of N-acetylcysteine (NAC) or blocking the activity of Nox, a protein leading to the formation of ROS, with diphenylene iodonium (DPI) inhibits the responses of BMM cells to RANKL, including ROS production, activation of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein (MAP) kinase, and extracellular signal regulated kinase (ERK), and osteoclast differentiation. Moreover, both RANKL mediated ROS production and osteoclast differentiation were completely blocked in precursors depleted of Nox1 activity by RNA interference or by expressing a dominant-negative mutant of Rac1. Together, these results indicate that ROSs act as an intracellular signal mediator for osteoclast differentiation. PMID- 15817679 TI - Early molecular response to posttransplantation imatinib determines outcome in MRD+ Philadelphia-positive acute lymphoblastic leukemia (Ph+ ALL). AB - In adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL), minimal residual disease (MRD) after stem cell transplantation (SCT) is associated with a relapse probability exceeding 90%. Starting imatinib in the setting of MRD may decrease this high relapse rate. In this prospective multicenter study, 27 Ph+ ALL patients received imatinib upon detection of MRD after SCT. Bcr-abl transcripts became undetectable in 14 (52%) of 27 patients, after a median of 1.5 months (0.9-3.7 months) ((early)CR(mol)). All patients who achieved an (early)CR(mol) remained in remission for the duration of imatinib treatment; 3 patients relapsed after imatinib was discontinued. Failure to achieve polymerase chain reaction (PCR) negativity shortly after starting imatinib predicted relapse, which occurred in 12 (92%) of 13 patients after a median of 3 months. Disease-free survival (DFS) in (early)CR(mol) patients is 91% +/- 9% and 54% +/- 21% after 12 and 24 months, respectively, compared with 8% +/- 7% after 12 months in patients remaining MRD+ (P < .001). In conclusion, approximately half of patients with Ph+ ALL receiving imatinib for MRD positivity after SCT experience prolonged DFS, which can be anticipated by the rapid achievement of a molecular complete remission (CR). Continued detection of bcr abl transcripts after 2 to 3 months on imatinib identifies patients who will ultimately experience relapse and in whom additional or alternative antileukemic treatment should be initiated. PMID- 15817680 TI - Letter to authors and readers regarding the new NIH policy on public access. PMID- 15817681 TI - On the molecular origins of the chronic myeloproliferative disorders: it all makes sense. PMID- 15817682 TI - Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. AB - Hepatic transdifferentiation of bone marrow cells has been previously demonstrated by intravenous administration of donor cells, which may recirculate to the liver after undergoing proliferation and differentiation in the recipient's bone marrow. In the present study, to elucidate which cellular components of human bone marrow more potently differentiate into hepatocytes, we fractionated human bone marrow cells into mesenchymal stem cells (MSCs), CD34+ cells, and non-MSCs/CD34- cells and examined them by directly xenografting to allylalcohol (AA)-treated rat liver. Hepatocyte-like cells, as revealed by positive immunostaining for human-specific alpha-fetoprotein (AFP), albumin (Alb), cytokeratin 19 (CK19), cytokeratin 18 (CK18), and asialoglycoprotein receptor (AGPR), and by reverse transcription-polymerase chain reaction (RT-PCR) for expression of AFP and Alb mRNA, were observed only in recipient livers with MSC fractions. Cell fusion was not likely involved since both human and rat chromosomes were independently identified by fluorescence in situ hybridization (FISH). The differentiation appeared to follow the process of hepatic ontogeny, reprogramming of gene expression in the genome of MSCs, as evidenced by expression of the AFP gene at an early stage and the albumin gene at a later stage. In conclusion, we have demonstrated that MSCs are the most potent component in hepatic differentiation, as revealed by directly xenografting into rat livers. PMID- 15817683 TI - Impaired interferon-gamma production as a consequence of STAT4 deficiency after autologous hematopoietic stem cell transplantation for lymphoma. AB - Production of interferon gamma (IFN-gamma) is critical for optimal antitumor immunotherapy in several preclinical animal models. Interleukin-12 (IL-12) induced IFN-gamma production is markedly defective after autologous stem cell transplantation. Quantitative deficiency in CD4 T cells, relative increase in CD25+CD4+ T cells, and bias toward T helper 2 (Th2) differentiation are not the primary mechanisms of defective IFN-gamma production. IL-12 receptor beta1 (IL 12Rbeta1) and IL-12Rbeta2 are expressed at equivalent or higher levels on posttransplantation patient peripheral blood mononuclear cells (PBMCs) as compared with control PBMCs. IL-12-induced tyrosine phosphorylation of signal transducer and activator of transcription 4 (STAT4) was undetectable or barely detectable in posttransplantation patient PBMCs, whereas IL-4-induced tyrosine phosphorylation of STAT6 did not differ in posttransplantation patient and control PBMCs. Levels of STAT4 protein were decreased by 97% in posttransplantation patient PBMCs. Levels of STAT4 mRNA were also significantly decreased in posttransplantation patient PBMCs. Incubation with IL-12 and IL-18 in combination partially reversed the defective IFN-gamma production by posttransplantation patient PBMCs. IFN-gamma production in response to IL-12 plus IL-18 did not require increased expression of STAT4 but was dependent on the activity of p38 mitogen-activated protein kinase (MAPK). These results indicate that defective IFN-gamma production is due to an intrinsic deficiency in STAT4 expression by posttransplantation patient lymphocytes and suggest strategies for circumventing this deficiency in cancer immunotherapy. PMID- 15817684 TI - Defective Vav expression and impaired F-actin reorganization in a subset of patients with common variable immunodeficiency characterized by T-cell defects. AB - Common variable immunodeficiency (CVID) is a primary immune disorder characterized by impaired antibody production, which is in many instances secondary to defective T-cell function (T-CVID). We have previously identified a subset of patients with T-CVID characterized by defective T-cell receptor (TCR) dependent protein tyrosine phosphorylation. In these patients, ZAP-70 fails to be recruited to the TCR as the result of impaired CD3zeta phosphorylation, which is, however, not dependent on defective Lck expression or activity. Here we show that neither Fyn nor CD45 is affected in these patients. On the other hand, T-CVID T cells show dramatic defects in the Vav/Rac pathway controlling F-actin dynamics. A significant deficiency in Vav protein was indeed observed; in 3 of 4 patients with T-CVID, it was associated with reduced VAV1 mRNA levels. The impairment in Vav expression correlated with defective F-actin reorganization in response to TCR/CD28 co-engagement. Furthermore, TCR/CD28-dependent up-regulation of lipid rafts at the cell surface, which requires F-actin dynamics, was impaired in these patients. The actin cytoskeleton defect could be reversed by reconstitution of Vav1 expression in the patients' T cells. Results demonstrate an essential role of Vav in human T cells and strongly suggest Vav insufficiency in T-CVID. PMID- 15817685 TI - Reverse recruitment: the Nup84 nuclear pore subcomplex mediates Rap1/Gcr1/Gcr2 transcriptional activation. AB - The recruitment model for gene activation presumes that DNA is a platform on which the requisite components of the transcriptional machinery are assembled. In contrast to this idea, we show here that Rap1/Gcr1/Gcr2 transcriptional activation in yeast cells occurs through a large anchored protein platform, the Nup84 nuclear pore subcomplex. Surprisingly, Nup84 and associated subcomplex components activate transcription themselves in vivo when fused to a heterologous DNA-binding domain. The Rap1 coactivators Gcr1 and Gcr2 form an important bridge between the yeast nuclear pore complex and the transcriptional machinery. Nucleoporin activation may be a widespread eukaryotic phenomenon, because it was first detected as a consequence of oncogenic rearrangements in acute myeloid leukemia and related syndromes in humans. These chromosomal translocations fuse a homeobox DNA-binding domain to the human homolog (hNup98) of a transcriptionally active component of the yeast Nup84 subcomplex. We conclude that Rap1 target genes are activated by moving to contact compartmentalized nuclear assemblages, rather than through recruitment of the requisite factors to chromatin by means of diffusion. We term this previously undescribed mechanism "reverse recruitment" and discuss the possibility that it is a central feature of eukaryotic gene regulation. Reverse recruitment stipulates that activators work by bringing the DNA to an nuclear pore complex-tethered platform of assembled transcriptional machine components. PMID- 15817686 TI - Stress-induced enhancement of leukocyte trafficking into sites of surgery or immune activation. AB - Effective immunoprotection requires rapid recruitment of leukocytes into sites of surgery, wounding, infection, or vaccination. In contrast to immunosuppressive chronic stressors, short-term acute stressors have immunoenhancing effects. Here, we quantify leukocyte infiltration within a surgical sponge to elucidate the kinetics, magnitude, subpopulation, and chemoattractant specificity of an acute stress-induced increase in leukocyte trafficking to a site of immune activation. Mice acutely stressed before sponge implantation showed 200-300% higher neutrophil, macrophage, natural killer cell, and T cell infiltration than did nonstressed animals. We also quantified the effects of acute stress on lymphotactin- (LTN; a predominantly lymphocyte-specific chemokine), and TNF-alpha (a proinflammatory cytokine) stimulated leukocyte infiltration. An additional stress-induced increase in infiltration was observed for neutrophils, in response to TNF-alpha, macrophages, in response to TNF-alpha and LTN, and natural killer cells and T cells in response to LTN. These results show that acute stress initially increases trafficking of all major leukocyte subpopulations to a site of immune activation. Tissue damage-, antigen-, or pathogen-driven chemoattractants subsequently determine which subpopulations are recruited more vigorously. Such stress-induced increases in leukocyte trafficking may enhance immunoprotection during surgery, vaccination, or infection, but may also exacerbate immunopathology during inflammatory (cardiovascular disease or gingivitis) or autoimmune (psoriasis, arthritis, or multiple sclerosis) diseases. PMID- 15817687 TI - pFind: a novel database-searching software system for automated peptide and protein identification via tandem mass spectrometry. AB - SUMMARY: Research in proteomics requires powerful database-searching software to automatically identify protein sequences in a complex protein mixture via tandem mass spectrometry. In this paper, we describe a novel database-searching software system called pFind (peptide/protein Finder), which employs an effective peptide scoring algorithm that we reported earlier. The pFind server is implemented with the C++ STL, .Net and XML technologies. As a result, high speed and good usability of the software are achieved. PMID- 15817688 TI - twilight; a Bioconductor package for estimating the local false discovery rate. AB - twilight is a Bioconductor compatible package for analysing the statistical significance of differentially expressed genes. It is based on the concept of the local false discovery rate (FDR), a generalization of the frequently used global FDR. twilight implements the heuristic search algorithm for estimating the local FDR introduced in our earlier work. In addition to the raw significance measures, it produces diagnostic plots, which provide insight into the extent of differential expression across genes. AVAILABILITY: http://www.bioconductor.org CONTACT: stefanie.scheid@molgen.mpg.de SUPPLEMENTARY INFORMATION: Please visit our software webpage on http://compdiag.molgen.mpg.de/software. PMID- 15817689 TI - Rapid and selective surveillance of Arabidopsis thaliana genome annotations with Centrifuge. AB - Centrifuge is a user-friendly system to simultaneously access Arabidopsis gene annotations and intra- and inter-organism sequence comparison data. The tool allows rapid retrieval of user-selected data for each annotated Arabidopsis gene providing, in any combination, data on the following features: predicted protein properties such as mass, pI, cellular location and transmembrane domains; SWISS PROT annotations; Interpro domains; Gene Ontology records; verified transcription; BLAST matches to the proteomes of A.thaliana, Oryza sativa (rice), Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. The tool lends itself particularly well to the rapid analysis of contigs or of tens or hundreds of genes identified by high-throughput gene expression experiments. In these cases, a summary table of principal predicted protein features for all genes is given followed by more detailed reports for each individual gene. Centrifuge can also be used for single gene analysis or in a word search mode. AVAILABILITY: http://centrifuge.unil.ch/ CONTACT: edward.farmer@unil.ch. PMID- 15817690 TI - Convergent Island Statistics: a fast method for determining local alignment score significance. AB - MOTIVATION: Background distribution statistics for profile-based sequence alignment algorithms cannot be calculated analytically, and hence such algorithms must resort to measuring the significance of an alignment score by assessing its location among a distribution of background alignment scores. The Gumbel parameters that describe this background distribution are usually pre-computed for a limited number of scoring systems, gap schemes, and sequence lengths and compositions. The use of such look-ups is known to introduce errors, which compromise the significance assessment of a remote homology relationship. One solution is to estimate the background distribution for each pair of interest by generating a large number of sequence shuffles and use the distribution of their scores to approximate the parameters of the underlying extreme value distribution. This is computationally very expensive, as a large number of shuffles are needed to precisely estimate the score statistics. RESULTS: Convergent Island Statistics (CIS) is a computationally efficient solution to the problem of calculating the Gumbel distribution parameters for an arbitrary pair of sequences and an arbitrary set of gap and scoring schemes. The basic idea behind our method is to recognize the lack of similarity for any pair of sequences early in the shuffling process and thus save on the search time. The method is particularly useful in the context of profile-profile alignment algorithms where the normalization of alignment scores has traditionally been a challenging task. CONTACT: aleksandar@eidogen.com SUPPLEMENTARY INFORMATION: http://www.eidogen-sertanty.com/Documents/convergent_island_stats_sup.pdf. PMID- 15817691 TI - Use of multiple profiles corresponding to a sequence alignment enables effective detection of remote homologues. AB - MOTIVATION: Position specific scoring matrices (PSSMs) corresponding to aligned sequences of homologous proteins are commonly used in homology detection. A PSSM is generated on the basis of one of the homologues as a reference sequence, which is the query in the case of PSI-BLAST searches. The reference sequence is chosen arbitrarily while generating PSSMs for reverse BLAST searches. In this work we demonstrate that the use of multiple PSSMs corresponding to a given alignment and variable reference sequences is more effective than using traditional single PSSMs and hidden Markov models. RESULTS: Searches for proteins with known 3-D structures have been made against three databases of protein family profiles corresponding to known structures: (1) One PSSM per family; (2) multiple PSSMs corresponding to an alignment and variable reference sequences for every family; and (3) hidden Markov models. A comparison of the performances of these three approaches suggests that the use of multiple PSSMs is most effective. CONTACT: ns@mbu.iisc.ernet.in. PMID- 15817692 TI - Stochastic reaction-diffusion simulation with MesoRD. AB - MesoRD is a tool for stochastic simulation of chemical reactions and diffusion. In particular, it is an implementation of the next subvolume method, which is an exact method to simulate the Markov process corresponding to the reaction diffusion master equation. AVAILABILITY: MesoRD is free software, written in C++ and licensed under the GNU general public license (GPL). MesoRD runs on Linux, Mac OS X, NetBSD, Solaris and Windows XP. It can be downloaded from http://mesord.sourceforge.net. CONTACT: johan.elf@icm.uu.se; johan.hattne@embl hamburg.de SUPPLEMENTARY INFORMATION: 'MesoRD User's Guide' and other documents are available at http://mesord.sourceforge.net. PMID- 15817693 TI - Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary. AB - MOTIVATION: High-throughput technologies such as DNA sequencing and microarrays have created the need for automated annotation of large sets of genes, including whole genomes, and automated identification of pathways. Ontologies, such as the popular Gene Ontology (GO), provide a common controlled vocabulary for these types of automated analysis. Yet, while GO offers tremendous value, it also has certain limitations such as the lack of direct association with pathways. RESULTS: We demonstrated the use of the KEGG Orthology (KO), part of the KEGG suite of resources, as an alternative controlled vocabulary for automated annotation and pathway identification. We developed a KO-Based Annotation System (KOBAS) that can automatically annotate a set of sequences with KO terms and identify both the most frequent and the statistically significantly enriched pathways. Results from both whole genome and microarray gene cluster annotations with KOBAS are comparable and complementary to known annotations. KOBAS is a freely available stand-alone Python program that can contribute significantly to genome annotation and microarray analysis. PMID- 15817694 TI - A protocol for building and evaluating predictors of disease state based on microarray data. AB - MOTIVATION: Microarray gene expression data are increasingly employed to identify sets of marker genes that accurately predict disease development and outcome in cancer. Many computational approaches have been proposed to construct such predictors. However, there is, as yet, no objective way to evaluate whether a new approach truly improves on the current state of the art. In addition no 'standard' computational approach has emerged which enables robust outcome prediction. RESULTS: An important contribution of this work is the description of a principled training and validation protocol, which allows objective evaluation of the complete methodology for constructing a predictor. We review the possible choices of computational approaches, with specific emphasis on predictor choice and reporter selection strategies. Employing this training-validation protocol, we evaluated different reporter selection strategies and predictors on six gene expression datasets of varying degrees of difficulty. We demonstrate that simple reporter selection strategies (forward filtering and shrunken centroids) work surprisingly well and outperform partial least squares in four of the six datasets. Similarly, simple predictors, such as the nearest mean classifier, outperform more complex classifiers. Our training-validation protocol provides a robust methodology to evaluate the performance of new computational approaches and to objectively compare outcome predictions on different datasets. PMID- 15817695 TI - Comment on 'Evaluation of the gene-specific dye bias in cDNA microarray experiments'. PMID- 15817696 TI - NodMutDB: a database for genes and mutants involved in symbiosis. AB - Functional genomics research is producing large amounts of data on the functions of individual genes related to symbiosis. We have developed a relational database, NodMutDB (Nodulation Mutant Database), to provide a comprehensive resource for depositing, organizing and retrieving information on symbiosis related genes, mutants and published literature. NodMutDB brings together new studies and existing mutant-based literature to facilitate our understanding of how genes function in symbiotic processes in both Rhizobia and their host plants. AVAILABILITY: http://nodmutdb.vbi.vt.edu CONTACT: cmao@vbi.vt.edu SUPPLEMENTARY INFORMATION: Database schema and data curation model are available at http://nodmutdb.vbi.vt.edu. PMID- 15817697 TI - Modeling and simulation of cancer immunoprevention vaccine. AB - We present an in silico model that simulates the immune system responses to tumor cells in naive and vaccinated mice. We have demonstrated the ability of this model to accurately reproduce the experimental results. MOTIVATION: In vivo experiments on HER-2/neu mice have shown the effectiveness of Triplex vaccine in the protection of mice from mammary carcinoma. Full protection was conferred using chronic (prophylactic) vaccination protocol while therapeutic vaccination was less efficient. Our in silico model was able to closely reproduce the effects of various vaccination protocols. This model is the first step towards the development of in silico experiments searching for optimal vaccination protocols. RESULTS: In silico experiments carried out on two large statistical samples of virtual mice showed very good agreements with in vivo experiments for all experimental vaccination protocols. They also show, as supported by in vivo experiments, that the humoral response is fundamental in controlling the tumor growth and therefore suggest the selection and timing of experiments for measuring the activity of T cells. CONTACT: francesco@dmi.unict.it SUPPLEMENTARY INFORMATION: http://www.dmi.unict.it/CIG/suppdata_bioinf.html. PMID- 15817698 TI - A boosting approach for motif modeling using ChIP-chip data. AB - MOTIVATION: Building an accurate binding model for a transcription factor (TF) is essential to differentiate its true binding targets from those spurious ones. This is an important step toward understanding gene regulation. RESULTS: This paper describes a boosting approach to modeling TF-DNA binding. Different from the widely used weight matrix model, which predicts TF-DNA binding based on a linear combination of position-specific contributions, our approach builds a TF binding classifier by combining a set of weight matrix based classifiers, thus yielding a non-linear binding decision rule. The proposed approach was applied to the ChIP-chip data of Saccharomyces cerevisiae. When compared with the weight matrix method, our new approach showed significant improvements on the specificity in a majority of cases. PMID- 15817699 TI - Angiotensin II increases human monocyte matrix metalloproteinase-1 through the AT2 receptor and prostaglandin E2: implications for atherosclerotic plaque rupture. AB - Angiotensin II (Ang II)-mediated hypertension increases the risk for acute coronary syndrome, a consequence of atherosclerotic plaque rupture, which may be caused by matrix metalloproteinases (MMPs). Here, we show that human primary monocytes stimulated with tumor necrosis factor alpha (TNF-alpha) and granulocyte macrophage-colony stimulating factor (GM-CSF) release Ang II, which is an integral component of the signal transduction pathway that leads to MMP-1 production. An Ang II-mediated increase in MMP-1 synthesis occurred only in conjunction with cytokine stimulation. Moreover, Ang II mediated its effect through the Ang II type 2 (AT(2)) receptor, as demonstrated by enhancement of MMP 1 production by an AT(2) agonist, CGP-42112A, and inhibition of MMP-1 production by PD1233319, an AT(2) antagonist. Additionally, exogenous Ang II caused a significant enhancement in MMP-1 production by cytokine-stimulated monocytes, and the most effective enhancement occurrred when Ang II was added 6 h after stimulation. Furthermore, Ang II and the AT(2) agonist increased prostaglandin E(2) (PGE(2)), which in turn mediated the increase in MMP-1, as shown by the inhibition of MMP-1 by indomethacin or aspirin. In contrast, the AT(2) antagonist inhibited the PGE(2) production induced by TNF-alpha and GM-CSF. Ang II, through its interaction with the AT(2) receptor, has a central role in mediating the PGE(2)-dependent production of MMP-1 by monocytes stimulated with TNF-alpha and GM-CSF. These observations provide insight into the association between hypertension and acute coronary syndrome and a possible mechanism by which Ang converting enzyme inhibitor and aspirin may reduce the risk for heart attacks. PMID- 15817700 TI - Mucosal application of plasmid-encoded IL-15 sustains a highly protective anti Herpes simplex virus immunity. AB - In a DNA immunization against Herpes simplex virus (HSV), we examined the ability of plasmid-encoded interleukin-15 (pIL-15) to induce and maintain the mucosal B and T cell immune response. pIL-15 generated memory CD8(+) T cell responses that were threefold higher and mainly maintained in the spleen, but high levels of immunoglobulin A antibodies were induced and maintained long-term in the vaginal mucosa. Both of these enhanced components of the immune responses were recalled rapidly upon challenge with a lethal dose of HSV McKrae, affording protection in mice immunized with codelivery of pIL-15. Our results show for the first time that intranasal administration of pIL-15 along with plasmid-encoded glycoprotein B of HSV leads to enhancement of primary and memory CD8(+) T cell responses as well as humoral immune response. Therefore, a mucosal immunization strategy that incorporates a potent cytokine such as IL-15 as an adjuvant might induce protective mucosal immune responses that constitute the initial barrier at mucosal portals of pathogen entry. PMID- 15817701 TI - TNFR1-induced sphingomyelinase activation modulates TCR signaling by impairing store-operated Ca2+ influx. AB - Tumor necrosis factor alpha (TNF-alpha) is a potent, pleiotrophic cytokine, which is proinflammatory but can also suppress T lymphocyte function. In chronic inflammatory disease such as rheumatoid arthritis, exposure of T cells to TNF alpha alters their ability to mount a response by modulating the T cell receptor (TCR) signaling pathway, but the mechanisms involved remain obscure. Here, we investigated the specific role of TNF receptor 1 (TNFR1) signaling in the modulation of the TCR signaling pathway. We observed a down-regulation of the intracellular calcium ([Ca(2+)](i)) signal in Jurkat T cells after just 30 min exposure to TNF-alpha, and maximum suppression was reached after 3 h. This effect was transient, and signals returned to normal after 12 h. This depression of [Ca(2+)](i) was also observed in human CD4+ T lymphocytes. The change in Ca(2+) signal was related to a decrease in the plasma membrane Ca(2+) influx, which was apparent even when the TCR signal was bypassed using thapsigargin to induce a Ca(2+) influx. The role of TNF-alpha-induced activation of the sphingolipid cascade in this pathway was examined. The engagement of TNFR1 by TNF-alpha led to a time-dependent increase in acid sphingomyelinase (SMase; ASM) activity, corresponding with a decrease in cellular sphingomyelin. In parallel, there was an increase in cellular ceramide, which correlated directly with the decrease in the magnitude of the Ca(2+) response to phytohemagglutinin. Exogenous addition of SMase or ceramide mimicked the effects of TNFR1 signals on Ca(2+) responses in Jurkat T cells. Direct evidence for the activation of ASM in this pathway was provided by complete abrogation of the TNF-alpha-induced inhibition of the Ca(2+) influx in an ASM-deficient murine T cell line (OT-II(+/+)ASM(-/-)). This potent ability of TNF-alpha to rapidly modulate the TCR Ca(2+) signal via TNFR1-induced ASM activation can explain its suppressive effect on T cell function. This TNFR1 signaling pathway may play a role as an important regulator of T cell responses. PMID- 15817702 TI - Minocycline attenuates T cell and microglia activity to impair cytokine production in T cell-microglia interaction. AB - Minocycline, a tetracycline with anti-inflammatory properties, has been reported to down-regulate the activity of microglia, whose activation occurs in inflammatory and degenerative diseases of the central nervous system, such as multiple sclerosis and Alzheimer's disease. In these disorders, a T cell component is also evident, and we have demonstrated previously that the interaction of activated T cells with microglia led to the substantial increase in tumor necrosis factor alpha (TNF-alpha) levels. Here, we report that minocycline decreases TNF-alpha levels produced in human T cell-microglia interaction. This effect is mediated by a direct action of minocycline on the activated T cells and on microglia, which resulted in the decreased ability of T cells to contact microglia. In correspondence, minocycline decreased the expression on T cells of the CD40 ligand (CD40L), a key molecule regulating the contact-mediated interaction of T cells with microglia. These results demonstrate that the mechanism of action of minocycline involves not only microglia but also T cells and their subsequent activation of microglia. The capacity of minocycline to down-regulate CD40L on T cells may provide a new means to target the CD40 CD40L pathway, which regulates several inflammatory processes. PMID- 15817703 TI - Asymmetrical protein kinase A activity establishes neutrophil cytoskeletal polarity and enables chemotaxis. AB - Neutrophil chemotaxis requires precise spatial organization of the actin cytoskeleton and integrin activation to polarize the cell and enable migration. Protein kinase A (PKA) activity regulates integrin activation and actin cytoskeletal organization, suggesting that PKA is a key element in the mechanism regulating neutrophil chemotaxis. Our hypothesis is that asymmetrical PKA activity is critical for establishing neutrophil adhesive and cytoskeletal polarity required for migration during chemotaxis. To test this hypothesis, we first determined that global treatment with the PKA inhibitor KT5720 decreased formylated Met-Leu-Phe (fMLF)-induced migration. The ability of PKA inhibitors to reduce migration correlated with increased overall beta2 integrin cell-surface expression, affinity activation, and cellular adhesion. We next determined whether asymmetrical PKA activity was sufficient to induce migration. Exposure to gradient of the PKA inhibitors KT5720 or H-89 or a stearated, cell-permeant peptide (St-Ht31), which inhibits PKA binding to anchorage proteins, stimulated neutrophil migration in a chemotaxis chamber. Global treatment with KT5720 abolished the ability of fMLF to polarize the neutrophil actin cytoskeleton. In contrast to global treatment with KT5720, a point source of KT5720 was sufficient to polarize the actin cytoskeleton. The ability of KT5720 and St-Ht31 to stimulate migration was abolished by pretreatment with the phosphatidylinositol-3 kinase (PI-3K) inhibitors wortmannin and LY294002. These data suggest that asymmetrical PKA activity is necessary and sufficient for actin cytoskeletal polarization and migration during neutrophil chemotaxis. In addition, our data suggest PI-3K is an effector of PKA during chemotaxis. PMID- 15817704 TI - Fas-mediated death and sensory adaptation limit the pathogenic potential of autoreactive T cells after strong antigenic stimulation. AB - The ability of autoreactive T cells to induce autoimmune pathology is dependent on their ability to respond to the level of autoantigen presented in the target organ. Emerging evidence suggests that at the population level, T cell sensitivity for self can be reduced by deletion of those cells bearing high affinity T cell receptors (TCRs) or by sensory adaptation of individual cells. Here, we have investigated the mechanisms that prevent the induction of experimental autoimmune encephalomyelitis (EAE) when myelin basic protein (MBP) reactive T cells are exposed to a strong, antigenic stimulus. Stimulation of MBP reactive TCR transgenic T cells with a superagonist peptide led to extensive activation-induced cell death (AICD) through Fas signaling. Using T cells lacking Fas, we found that disruption of this deletional mechanism only partially increased EAE in response to superagonist, failing to restore susceptibility to the level found in response to the wild-type MBP peptide. A significant fraction of the MBP-reactive T cells was able to avoid AICD in response to superagonist, but these cells had a reduced sensitivity for an antigen that correlated with elevated levels of CD5. Therefore, when TCR affinity is fixed, autoreactive T cell sensitivity can be shifted to below a threshold for harm by a combination of AICD and sensory adaptation. PMID- 15817705 TI - T cells from paroxysmal nocturnal haemoglobinuria (PNH) patients show an altered CD40-dependent pathway. AB - Paroxysmal nocturnal haemoglobinuria (PNH) is a haematopoiesis disorder characterized by the expansion of a stem cell bearing a somatic mutation in the phosphatidylinositol glycan-A (PIG-A) gene, which is involved in the biosynthesis of the glycosylphosphatidylinositol (GPI) anchor. A number of data suggest the inability of the PIG-A mutation to account alone for the clonal dominance of the GPI-defective clone and for the development of PNH. In this context, additional immune-mediated mechanisms have been hypothesized. We focused on the analysis of T lymphocytes in three PNH patients bearing a mixed GPI(+) and GPI(-) T cell population and showing a marked cytopenia. To analyze the biological mechanisms underlying the control of T cell homeostasis in PNH, we addressed the study of CD40-dependent pathways, suggested to be of crucial relevance for the control of autoreactive T cell clones. Our data revealed significant, functional alterations in GPI(+) and GPI(-) T cell compartments. In the GPI(-) T cells, severe defects in T cell receptor-dependent proliferation, interferon-gamma production, CD25, CD54, and human leukocyte antigen-DR surface expression were observed. By contrast, GPI(+) T lymphocytes showed a significant increase of all these parameters, and the analysis of CD40-dependent pathways revealed a functional persistence of CD154 expression on the CD48(+)CD4(+) lymphocytes. The alterations of the GPI(+) T cell subset could be involved in the biological mechanisms underlying PNH pathogenesis. PMID- 15817706 TI - HLA-DO transduced in human monocyte-derived dendritic cells modulates MHC class II antigen processing. AB - Through the regulation of human leukocyte antigen (HLA)-DM (DM) in B cells, HLA DO (DO) modulates positively or negatively the presentation of specific peptides. Transduction of DO into human blood monocyte-derived dendritic cells (MoDC) has been proposed as a mean of modifying the peptide repertoire of major histocompatibility complex class II molecules. However, maturation of DC induced by inflammatory stimuli or possibly the adenoviral vector itself triggers acidification of vesicles and shuts down transcription of the class II transactivator gene as well as de novo biosynthesis of class II-related molecules and DM activity. In these conditions, it is unclear that transduced DO could alter the peptide repertoire. Our Western blot and reverse transcriptase polymerase chain reaction analyses revealed that human DC derived from blood monocytes express small amounts of DOalpha. Transduction of DObeta alone resulted in the accumulation of a small pool of DO in DM(+) CD63(+) vesicles and at the plasma membrane of mature DC. The cell-surface increase in class II-associated invariant chain peptide (CLIP)/class II complexes is in line with an inhibitory role of DO on DM. Cotransduction of DOalpha and DObeta only slightly increased CLIP and DO levels at the cell surface. Together with the fact that a large fraction of transduced DO remains in the endoplasmic reticulum, this suggests that DM is limiting in these conditions. DO expression did not affect a mixed lymphocyte reaction but reduced presentation of the exogenous gp100 antigen to a specific T cell clone. These results show that transduced DO modulates antigen presentation in human mature MoDC, evoking the possible use of this chaperone for immunotherapy. PMID- 15817707 TI - Leukocyte apoptosis and its significance in sepsis and shock. AB - Sepsis and multiple organ failure continue to be significant problems among trauma, burn, and the critically ill patient population. Thus, a number of laboratories have focused on understanding the role of altered apoptotic cell death in contributing to immune and organ dysfunction seen in sepsis and shock. Immune cells that undergo altered apoptotic changes include neutrophils, macrophages, dendritic cells, as well as various lymphocyte populations. Evidence of epithelial as well as endothelial cell apoptotic changes has also been reported. Although mediators such as steroids, tumor necrosis factor, nitric oxide, C5a, and Fas ligand (FasL) appear to contribute to the apoptotic changes, their effects are tissue- and cell population-selective. As inhibiting Fas-FasL signaling (e.g., gene deficiency, Fas fusion protein, or Fas short interfering RNA administration), caspase inhibition (caspase mimetic peptides), and/or the overexpression of downstream antiapoptotic molecules (e.g., Bcl-2, Akt) improve survival of septic mice, it not only demonstrates the pathological significance of this process but points to novel targets for the treatment of sepsis. PMID- 15817708 TI - Augmented IL-10 production and redox-dependent signaling pathways in glucose-6 phosphate dehydrogenase-deficient mouse peritoneal macrophages. AB - Glucose-6-phosphate dehydrogenase (G6PD) supports cellular antioxidant pathways. G6PD deficiency is associated with malaria protection but was shown to worsen the clinical course to injury. This study tested whether G6PD deficiency manifests in altered cytokine responses using peritoneal macrophages from a G6PD-deficient mouse model with a degree of defect similar to the common type A(-) human G6PD deficiency. Lipopolysaccharide (LPS)-induced interleukin (IL)-10 and IL-12 production was doubled in G6PD-deficient macrophages compared with wild-type (WT). Protein kinase C (PKC) activation by phorbol-ester prior to LPS resulted in a fivefold greater IL-10 production in G6PD-deficient macrophages compared with WT. Interferon-gamma treatment prior to LPS augmented IL-12 production in G6PD deficient and WT macrophages and partially inhibited IL-10 production by G6PD deficient macrophages. The antioxidants (N-acetyl-L-cysteine and glutathione ethyl-ester) blunted IL-10 and IL-12 production, indicating a role for oxidative stress in the observed response differences between deficient and WT macrophages. LPS-induced activation of nuclear factor-kappaB, cyclic adenosine monophosphate response element-binding protein, and specificity protein 3 was augmented in G6PD deficient cells compared with WT. The PKCdelta inhibitor Rottlerin inhibited IL 10 and IL-12 production at different 50% effective-dose concentrations between deficient and WT macrophages, indicating elevated PKCdelta activity in deficient cells. This study reveals that activated G6PD-deficient macrophages display an augmented production of cytokines with a prominent impact on IL-10 production. The altered cytokine responses are associated with augmented activation of redox dependent transcription factors and PKCdelta. Alterations in signaling pathways and associated changes in cytokine production may play a role in modulating the inflammatory responses following bacterial or malarial infections in G6PD deficiency. PMID- 15817709 TI - Plasma levobupivacaine concentrations following scalp block in patients undergoing awake craniotomy. AB - BACKGROUND: Levobupivacaine is an effective local anaesthetic agent for nerve blockade with less systemic toxicity than racemic bupivacaine. The safety and efficacy of levobupivacaine for scalp blockade during awake craniotomy have not been addressed previously. METHODS: Serial arterial plasma levobupivacaine concentrations following scalp blockade were measured to 2 h in 10 patients booked for awake craniotomy for epilepsy or tumour surgery. Bilateral scalp blockade providing surgical anaesthesia was achieved with a mean dose of 177 mg (2.5 mg kg(-1), range 1.6-3.2 mg kg(-1)) of levobupivacaine (0.5%, 5 mg ml(-1)) with epinephrine (5 microg ml(-1)) added immediately before the block insertion. RESULTS: The maximum measured plasma levobupivacaine concentration was 1.58 (0.44) microg ml(-1) [mean (SD)] with a mean time to peak plasma concentration of 12 (4) min. There were no episodes in any of the 10 patients of symptoms or signs suggestive of either CNS or CVS toxicity. CONCLUSIONS: This study demonstrated a relatively rapid rise of plasma levobupivacaine concentration without evidence of cardiovascular or central nervous system sequelae in a sample population of patients who may be particularly prone to perioperative seizures. PMID- 15817710 TI - Incidence of lower thoracic ligamentum flavum midline gaps. AB - BACKGROUND: Lower thoracic epidural anaesthesia and analgesia (EDA) has gained increasing importance in perioperative pain therapy. The loss-of-resistance technique used to identify the epidural space is thought to rely on the penetration of the ligamentum flavum. Investigations at the cervical and lumbar regions have demonstrated that the ligamentum flavum frequently exhibits incomplete fusion at different vertebral levels. Therefore, the aim of this study was to directly investigate the incidence of lower thoracic ligamentum flavum midline gaps in embalmed cadavers. METHODS: Vertebral column specimens were obtained from 47 human cadavers. Ligamentum flavum midline gaps were recorded between the vertebral levels T6 and L1. RESULTS: The incidence of midline gaps/number of viable specimens at the following levels was: T6-7: 2/45 (4.4%), T7-8: 1/47 (2.1%), T8-9: 2/45 (4.4%), T9-10: 7/39 (17.9%), T10-11: 12/34 (35.2%), T11-12: 10/35 (28.5%), T12/L1: 6/38 (15.8%). CONCLUSIONS: In the present study we have determined the frequency of lower thoracic ligamentum flavum midline gaps. Gaps are less frequent than at cervical levels, but more frequent than at lumbar levels. Peak incidence was found in the region between T10 and T12. Using a strict midline approach, one cannot therefore rely on the ligamentum flavum to impede entering the epidural space in all patients. PMID- 15817711 TI - Methodological approaches to anaesthetists' workload in the operating theatre. AB - This review examines the basic concepts of workload and methods of measuring them. The components of anaesthetists' operating room activities, and the factors contributing to workload, are analysed using an ergonomic-based model for technological environments. The available evidence on the relationship between workload and training and supervision is presented and the effect of workload on the quality and safety of anaesthetic care is reviewed. There is, as yet, only a small body of work examining workload and its effects in anaesthesia. While studies have identified the general pattern of workload in relation to the different stages of the perioperative period, measurement, particularly of overlapping tasks, is still evolving. It is clear, however, that induction and emergence are the most intense periods of both practical and 'non-technical' aspects of work. Allocation of attention to a range of tasks simultaneously is a key characteristic of anaesthetic practice. Experienced staff appear to show 'spare capacity' in performance during routine cases, which, we suggest, allows them an attentional 'safety margin' should adverse events occur. The effects of production pressure and mental 'overload' remain speculative and so practical recommendations for anaesthetic staffing, both in terms of numbers and matching skills to surgical demand, cannot be made. The potential for delegation of tasks, for instance to non-physician anaesthetists, can also not be made on evidence based grounds. Strategies for active management of workload may be useful in practice. PMID- 15817712 TI - Comparison of the tyrosine aminotransferase cDNA and genomic DNA sequences of normal mink and mink affected with tyrosinemia type II. AB - Type II tyrosinemia, designated Richner-Hanhart syndrome in humans, is a hereditary metabolic disorder with autosomal recessive inheritance characterized by a deficiency of tyrosine aminotransferase activity. Mutations occur in the human tyrosine aminotransferase gene, resulting in high levels of tyrosine and disease. Type II tyrosinemia occurs in mink, and our hypothesis was that it would also be associated with mutation(s) in the tyrosine aminotransferase gene. Therefore, the transcribed cDNA and the genomic tyrosine aminotransferase gene were sequenced from normal and affected mink. The gene extended over 11.9 kb and had 12 exons coding for a predicted 454-amino-acid protein with 93% homology with human tyrosine aminotransferase. FISH analysis mapped the gene to chromosome 8 using the Mandahl and Fredga (1975) nomenclature and chromosome 5 using the Christensen et al. (1996) nomenclature. The hypothesis was rejected because sequence analysis disclosed no mutations in either cDNA or introns that were associated with affected mink. This suggests that an unlinked gene regulatory mutation may be the cause of tyrosinemia in mink. PMID- 15817713 TI - Attempted replication of reported chronic obstructive pulmonary disease candidate gene associations. AB - Case-control studies have successfully identified many significant genetic associations for complex diseases, but lack of replication has been a criticism of case-control genetic association studies in general. We selected 12 candidate genes with reported associations to chronic obstructive pulmonary disease (COPD) and genotyped 29 polymorphisms in a family-based study and in a case-control study. In the Boston Early-Onset COPD Study families, significant associations with quantitative and/or qualitative COPD-related phenotypes were found for the tumor necrosis factor (TNF)-alpha -308G>A promoter polymorphism (P < 0.02), a coding variant in surfactant protein B (SFTPB Thr131Ile) (P = 0.03), and the (GT)(31) allele of the heme oxygenase (HMOX1) promoter short tandem repeat (P = 0.02). In the case-control study, the SFTPB Thr131Ile polymorphism was associated with COPD, but only in the presence of a gene-by-environment interaction term (P = 0.01 for both main effect and interaction). The 30-repeat, but not the 31 repeat, allele of HMOX1 was associated (P = 0.04). The TNF -308G>A polymorphism was not significant. In addition, the microsomal epoxide hydrolase "fast" allele (EPHX1 His139Arg) was significantly associated in the case-control study (P = 0.03). Although some evidence for replication was found for SFTPB and HMOX1, none of the previously published COPD genetic associations was convincingly replicated across both study designs. PMID- 15817715 TI - Brain stem mechanisms underlying acupuncture modality-related modulation of cardiovascular responses in rats. AB - The present study was designed to investigate brain stem responses to manual acupuncture (MA) and electroacupuncture (EA) at different frequencies at pericardial P (5-6) acupoints located over the median nerve. Activity of premotor sympathetic cardiovascular neurons in the rostral ventral lateral medulla (rVLM) was recorded during stimulation of visceral and somatic afferents in ventilated anesthetized rats. We stimulated either the splanchnic nerve at 2 Hz (0.1-0.4 mA, 0.5 ms) or the median nerve for 30 s at 2, 10, 20, 40, or 100 Hz using EA (0.3 0.5 mA, 0.5 ms) or at approximately 2 Hz with MA. Twelve of 18 cells responsive to splanchnic and median nerve stimulation could be antidromically driven from the intermediolateral columns of the thoracic spinal cord, T2-T4, indicating that they were premotor sympathetic neurons. All 18 neurons received baroreceptor input, providing evidence of their cardiovascular sympathoexcitatory function. Evoked responses during stimulation of the splanchnic nerve were inhibited by 49 +/- 6% (n = 7) with EA and by 46 +/- 4% (n = 6) with MA, indicating that the extent of inhibitory effects of the two modalities were similar. Inhibition lasted for 20 min after termination of EA or MA. Cardiovascular premotor rVLM neurons responded to 2-Hz electrical stimulation at P 5-6 and to a lesser extent to 10-, 20-, 40-, and 100-Hz stimulation (53 +/- 10, 16 +/- 2, 8 +/- 2, 2 +/- 1, and 0 +/- 0 impulses/30 stimulations, n = 7). These results indicate that rVLM premotor sympathetic cardiovascular neurons that receive convergent input from the splanchnic and median nerves during low-frequency EA and MA are inhibited similarly for prolonged periods by low-frequency MA and EA. PMID- 15817714 TI - Polyspecific cation transporters mediate luminal release of acetylcholine from bronchial epithelium. AB - In airway epithelia, non-neuronal cholinergic regulations have been described; however, the route for acetylcholine (ACh) release has not been verified. To investigate whether organic cation transporters (OCTs) serve this function, we studied the expression of OCTs in airway epithelia and their capability to translocate ACh. Using immunohistochemistry in rats and humans, OCT1, OCT2, and OCT3 were localized to the luminal membrane of ciliated epithelial cells. In humans, OCT2 showed the strongest expression in the luminal membrane. We expressed the OCT isoforms in oocytes of Xenopus laevis and measured uptake and efflux of ACh. Tracer flux measurements showed that ACh is transported by OCT1 and OCT2 but not by OCT3. Two-electrode-voltage-clamp measurements revealed that OCT2 mediates electrogenic uptake and efflux of ACh. For ACh uptake by human OCT2, a K(M) value of approximately 0.15 mM was determined. At -50 mV, ACh efflux by human OCT2 was trans-inhibited by micromolar concentrations of the inhalational glucocorticoid budesonide, which is used in treatment of asthma (K(i) approximately 2.7 microM). The data show that OCT1 and OCT2 mediate luminal ACh release in human airways and suggest that ACh release is blocked after inhalation of budesonide. PMID- 15817716 TI - Comparison of the symptoms of exercise-induced muscle damage after an initial and repeated bout of plyometric exercise in men and boys. AB - The purpose of this study was to compare symptoms of exercise-induced muscle damage after an initial and repeated bout of plyometric exercise in men and boys. Ten boys (9-10 yr) and 10 men (20-29 yr) completed two bouts of eight sets of 10 plyometric jumps, 2 wk apart. Perceived soreness (0-10, visual analog scale), isometric strength of the quadriceps at six knee flexion angles, and countermovement jump and squat jump height were assessed before and at 30 min, 24 h, 48 h, and 72 h after each bout. All variables followed the expected patterns of change in men, with soreness peaking at 24-48 h (5.8 +/- 1.7) and decrements in muscle function peaking at 30 min after the first bout (73-85% of baseline scores). Symptoms remained for 72 h after the first bout in men. In boys, symptoms were much less severe and peaked at 30 min (visual analog scale = 2.1 +/ 1.8, functional decrements 87-92% of baseline) and, with the exception of soreness, returned to baseline after 24 h. After the second bout of plyometric exercise, the level of soreness and decrements in countermovement jump, squat jump, and isometric strength were lower, although the effect was stronger in men, in all cases. The results of this study suggest that although children may experience symptoms of muscle damage after intensive plyometric exercise, they are much less severe. A prior bout of plyometric exercise also appears to provide children with some protection from soreness after a subsequent bout of plyometric exercise. Explanations for milder symptoms of exercise-induced muscle damage in children include greater flexibility leading to less overextension of sarcomeres during eccentric exercise, fewer fast-twitch muscle fibers, and greater and perhaps more varied habitual physical activity patterns. PMID- 15817717 TI - Developmental changes in airway and tissue mechanics in mice. AB - Most studies using mice to model human lung diseases are carried out in adults, although there is emerging interest in the effects of allergen, bacterial, and viral exposure early in life. This study aims to characterize lung function in BALB/c mice from infancy (2 wk) through to adulthood (8 wk). The low-frequency forced oscillation technique was used to obtain impedance data, partitioned into components representing airway resistance, tissue damping, tissue elastance, and hysteresivity (tissue damping/tissue elastance). Measurements were made at end expiratory pause (transrespiratory system pressure = 2 cmH2O) and during relaxed slow expiration from 20 to 0 cmH2O. Airway resistance decreased with age from 0.63 cmH2O x ml(-1) x s at 2 wk to 0.24 cmH2O x ml(-1) x s at 8 wk (P < 0.001). Both tissue damping and tissue elastance decreased with age (P < 0.001) from 2 to 5 wk, then plateaued through to 8 wk (P < 0.001). This pattern was seen both in measurements taken at end-expiratory pause and during expiration. There were no age-related changes seen in hysteresivity when measured at end-expiratory pause, but the pattern of volume dependence did differ with the age of the mice. These changes in respiratory mechanics parallel the reported structural changes of the murine lung from the postnatal period into adulthood. PMID- 15817718 TI - Tyrosine hydroxylase expression and activity in the rat brain: differential regulation after long-term intermittent or sustained hypoxia. AB - Tyrosine hydroxylase, a hypoxia-regulated gene, may be involved in tissue adaptation to hypoxia. Intermittent hypoxia, a characteristic feature of sleep apnea, leads to significant memory deficits, as well as to cortex and hippocampal apoptosis that are absent after sustained hypoxia. To examine the hypothesis that sustained and intermittent hypoxia induce different catecholaminergic responses, changes in tyrosine hydroxylase mRNA, protein expression, and activity were compared in various brain regions of male rats exposed for 6 h, 1 day, 3 days, and 7 days to sustained hypoxia (10% O(2)), intermittent hypoxia (alternating room air and 10% O(2)), or normoxia. Tyrosine hydroxylase activity, measured at 7 days, increased in the cortex as follows: sustained > intermittent > normoxia. Furthermore, activity decreased in the brain stem and was unchanged in other brain regions of sustained hypoxia-exposed rats, as well as in all regions from animals exposed to intermittent hypoxia, suggesting stimulus-specific and heterotopic catecholamine regulation. In the cortex, tyrosine hydroxylase mRNA expression was increased, whereas protein expression remained unchanged. In addition, significant differences in the time course of cortical Ser(40) tyrosine hydroxylase phosphorylation were present in the cortex, suggesting that intermittent and sustained hypoxia-induced enzymatic activity differences are related to different phosphorylation patterns. We conclude that long-term hypoxia induces site-specific changes in tyrosine hydroxylase activity and that intermittent hypoxia elicits reduced tyrosine hydroxylase recruitment and phosphorylation compared with sustained hypoxia. Such changes may not only account for differences in enzyme activity but also suggest that, with differential regional brain susceptibility to hypoxia, recruitment of different mechanisms in response to hypoxia will elicit region-specific modulation of catecholamine response. PMID- 15817719 TI - Mechanical stimulation of the plantar foot surface attenuates soleus muscle atrophy induced by hindlimb unloading in rats. AB - Unloading-induced muscle atrophy occurs in the aging population, bed-ridden patients, and astronauts. This study was designed to determine whether dynamic foot stimulation (DFS) applied to the plantar surface of the rat foot can serve as a countermeasure to soleus muscle atrophy normally observed in hindlimb unloaded (HU) rats. Forty-four mature (6 mo old), male Wistar rats were randomly assigned to ambulatory control, HU alone, HU with active DFS (i.e., plantar contact with active inflation), HU with passive DFS (i.e., plantar contact without active inflation), and HU while wearing a DFS boot with no plantar contact groups. Application of active DFS during HU significantly counteracted the atrophic response by preventing approximately 85% of the reduction in type I myofiber cross-sectional area (CSA) in the soleus while preventing approximately 57% of the reduction in type I myofiber CSA and 43% of the reduction in type IIA myofiber CSA of the medial gastrocnemius muscle. Wearing of a DFS boot without active inflation prevented myofiber atrophy in the soleus of HU animals in a fashion similar to that observed in HU animals that wore an actively inflated DFS boot. However, when a DFS boot without plantar surface contact was worn during HU, no significant protection from HU-induced myofiber atrophy was observed. These results illustrate that the application of mechanical foot stimulation to the plantar surface of the rat foot is an effective countermeasure to muscle atrophy induced by HU. PMID- 15817720 TI - Kinetics of .VO2 and femoral artery blood flow during heavy-intensity, knee extension exercise. AB - It has been suggested that, during heavy-intensity exercise, O(2) delivery may limit oxygen uptake (.VO2) kinetics; however, there are limited data regarding the relationship of blood flow and .VO2 kinetics for heavy-intensity exercise. The purpose was to determine the exercise on-transient time course of femoral artery blood flow (Q(leg)) in relation to .VO2 during heavy-intensity, single leg, knee-extension exercise. Five young subjects performed five to eight repeats of heavy-intensity exercise with measures of breath-by-breath pulmonary .VO2 and Doppler ultrasound femoral artery mean blood velocity and vessel diameter. The phase 2 time frame for .VO2 and Q(leg) was isolated and fit with a monoexponent to characterize the amplitude and time course of the responses. Amplitude of the phase 3 response was also determined. The phase 2 time constant for .VO2 of 29.0 s and time constant for Q(leg) of 24.5 s were not different. The change (Delta) in .VO2 response to the end of phase 2 of 0.317 l/min was accompanied by a DeltaQ(leg) of 2.35 l/min, giving a DeltaQ(leg)-to-Delta.VO2 ratio of 7.4. A slow component .VO2 of 0.098 l/min was accompanied by a further Q(leg) increase of 0.72 l/min (DeltaQ(leg)-to-Delta.VO2 ratio = 7.3). Thus the time course of Q(leg) was similar to that of muscle .VO2 (as measured by the phase 2 .VO2 kinetics), and throughout the on-transient the amplitude of the Q(leg) increase achieved (or exceeded) the Q(leg)-to-.VO2 ratio steady-state relationship (ratio approximately 4.9). Additionally, the .VO2 slow component was accompanied by a relatively large rise in Q(leg), with the increased O(2) delivery meeting the increased Vo(2). Thus, in heavy-intensity, single-leg, knee-extension exercise, the amplitude and kinetics of blood flow to the exercising limb appear to be closely linked to the .VO2 kinetics. PMID- 15817721 TI - Na+-K+-ATPase properties in rat heart and skeletal muscle 3 mo after coronary artery ligation. AB - This study was designed to determine whether chronic heart failure (CHF) results in changes in Na(+)-K(+)-ATPase properties in heart and skeletal muscles of different fiber-type composition. Adult rats were randomly assigned to a control (Con; n = 8) or CHF (n = 8) group. CHF was induced by ligation of the left main coronary artery. Examination of Na(+)-K(+)-ATPase activity (means +/- SE) 12 wk after the ligation measured, using the 3-O-methylfluorescein phosphatase assay (3 O-MFPase), indicated higher (P < 0.05) levels in soleus (Sol) (250 +/- 13 vs. 179 +/- 18 nmol.mg protein(-1).h(-1)) and lower (P < 0.05) levels in diaphragm (Dia) (200 +/- 12 vs. 272 +/- 27 nmol.mg protein(-1).h(-1)) and left ventricle (LV) (760 +/- 62 vs. 992 +/- 16 nmol.mg protein(-1).h(-1)) in CHF compared with Con, respectively. Na(+)-K(+)-ATPase protein content, measured by the [(3)H]ouabain binding technique, was higher (P < 0.05) in white gastrocnemius (WG) (166 +/- 12 vs. 135 +/- 7.6 pmol/g wet wt) and lower (P < 0.05) in Sol (193 +/- 20 vs. 260 +/ 8.6 pmol/g wet wt) and LV (159 +/- 10 vs. 221 +/- 10 pmol/g wet wt) in CHF compared with Con, respectively. Isoform content in CHF, measured by Western blot techniques, showed both increases (WG; P < 0.05) and decreases (Sol; P < 0.05) in alpha(1). For alpha(2), only increases [red gastrocnemius (RG), Sol, and Dia; P < 0.05] occurred. The beta(2)-isoform was decreased (LV, Sol, RG, and WG; P < 0.05) in CHF, whereas the beta(1) was both increased (WG and Dia; P < 0.05) and decreased (Sol and LV; P < 0.05). For beta(3), decreases (P < 0.05) in RG were observed in CHF, whereas no differences were found in Sol and WG between CHF and Con. It is concluded that CHF results in alterations in Na(+)-K(+)-ATPase that are muscle specific and property specific. Although decreases in Na(+)-K(+) ATPase content would appear to explain the lower 3-O-MFPase in the LV, such does not appear to be the case in skeletal muscles where a dissociation between these properties was observed. PMID- 15817722 TI - Long-term electromyographic activity in upper trapezius and low back muscles of women with moderate physical activity. AB - The habitual activity patterns of trapezius and postural back muscles (multifidus, iliocostalis, longissimus) of 23 female subjects with moderate physical activity were studied. Bilateral surface electromyographic (sEMG) recordings from start of work until bedtime were analyzed. The activity level was calibrated as percentage of root mean square-detected muscle activity at maximal voluntary contraction (EMG(max)). Sixty-six previous trapezius recordings of women with moderate physical activity were included in some analyses to pursue the full range of variation in trapezius activity. Twenty-six of these were recorded twice, separated by 16-28 mo. Median activity level and duration of periods with sEMG activity of <0.5% EMG(max) ("rest time"; only trapezius) and exceeding 2 ("burst time"), 10, 30, and 50% EMG(max) was determined. The trapezius median activity level ranged from 0.6 to 8.8% EMG(max), burst time from 9 to 84%, and rest time from 2 to 84%. The activity patterns of the back muscles showed similar large interindividual variation. Repeated trapezius recordings of the same subject showed high consistency; intraclass correlation coefficients ranged from 0.62 to 0.79 for different sEMG variables. Periods with high sEMG amplitude were of short duration; 7% of the trapezius recordings did not present time intervals (0.2-s duration) above 50% EMG(max). The activity patterns of the postural muscles, despite large interindividual variability, were distinctly different from activity patterns of upper and lower limb muscles reported by others (e.g., mean burst time 40-50 vs. 10-20%). We conclude that postural trunk muscles show idiosyncratic activity patterns with large interindividual variation. High-threshold motor units are activated to a very minor extent. PMID- 15817723 TI - Glycerol hyperhydration: physiological responses during cold-air exposure. AB - Hypohydration occurs during cold-air exposure (CAE) through combined effects of reduced fluid intake and increased fluid losses. Because hypohydration is associated with reduced physical performance, strategies for maintaining hydration during CAE are important. Glycerol ingestion (GI) can induce hyperhydration in hot and temperate environments, resulting in greater fluid retention compared with water (WI) alone, but it is not effective during cold water immersion. Water immersion induces a greater natriuresis and diuresis than cold exposure; therefore, whether GI might be effective for hyperhydration during CAE remains unknown. This study examined physiological responses, i.e., thermoregulatory, cardiovascular, renal, vascular fluid, and fluid-regulating hormonal responses, to GI in seven men during 4 h CAE (15 degrees C, 30% relative humidity). Subjects completed three separate, double-blind, and counterbalanced trials including WI (37 ml water/l total body water), GI (37 ml water/l total body water plus 1.5 g glycerol/l total body water), and no fluid. Fluids were ingested 30 min before CAE. Thermoregulatory responses to cold were similar during each trial. Urine flow rates were higher (P = 0.0001) with WI (peak 11.8 ml/min, SD 1.9) than GI (5.0 ml/min, SD 1.8), and fluid retention was greater (P = 0.0001) with GI (34%, SD 7) than WI (18%, SD 5) at the end of CAE. Differences in urine flow rate and fluid retention were the result of a greater free water clearance with WI. These data indicate glycerol can be an effective hyperhydrating agent during CAE. PMID- 15817724 TI - Respiratory system responsiveness in rabbits in vivo is reduced by prolonged continuous positive airway pressure. AB - Active, nonanesthetized, tracheotomized rabbits were subjected to continuous positive airway pressure (CPAP) for 4 days to determine the effects of chronic mechanical strain on lung and airway function. Rabbits were maintained for 4 days at a CPAP of 6 cmH(2)O (high CPAP), at a CPAP of 0 cmH(2)O (low CPAP), or without tracheostomy (no CPAP). After treatment with CPAP, changes in respiratory resistance in response to increasing concentrations of inhaled ACh were measured during mechanical ventilation to evaluate respiratory system responsiveness in vivo. Intraparenchymal bronchial segments were isolated from the lungs of all animals to evaluate airway smooth muscle responsiveness and bronchial compliance in vitro. Rabbits maintained for 4 days at high CPAP demonstrated significantly lower responsiveness to ACh compared with rabbits that were maintained at low CPAP or with no CPAP. Airways isolated from the lungs of animals subjected to the chronic application of high CPAP were also less responsive to ACh in vitro than the airways isolated from animals subjected to low CPAP or no CPAP. The persistence of the decreased responsiveness in the excised airway tissues suggests that the decreased respiratory system responsiveness observed in vivo results primarily from direct effects on the airways. The results demonstrate that the application of prolonged mechanical strain in vivo can reduce airway reactivity. PMID- 15817725 TI - ACTN3 and MLCK genotype associations with exertional muscle damage. AB - Strenuous exercise results in damage to skeletal muscle that is manifested in delayed muscle pain, prolonged strength loss, and increases in muscle proteins in the blood, especially creatine kinase (CK) and myoglobin (Mb). Some individuals experience profound changes in these variables in response to standard laboratory exercise or recreational activities. We proposed that variations in genes coding for two myofibrillar proteins [alpha-actinin 3 (ACTN3) and myosin light chain kinase (MLCK)] may explain the large variability in the response to muscle damaging exercise. We hypothesized that subjects with specific single nucleotide polymorphisms (SNPs) in ACTN3 and MLCK would show a greater loss in muscle strength and/or a greater increase in blood CK and Mb in response to eccentric exercise. Blood from 157 subjects who performed a standard elbow flexion eccentric exercise protocol was tested for association between genotypes of ACTN3 (1 SNP tested: R577X) and MLCK (2 SNPs tested: C49T and C37885A) and changes in blood CK and Mb and isometric strength. Subjects possessing the ACTN3-deficient genotype (XX) had lower baseline CK compared with the heterozygotes (P = 0.035). After the eccentric exercise, those subjects homozygous for the MLCK 49T rare allele had a significantly greater increase in CK and Mb (P < 0.01) compared with the heterozygotes, and those heterozygous for MLCK C37885A had a significantly greater increase in CK compared with the homozygous wild type (P < 0.05). There was only one subject homozygous for the rare MLCK 37885A allele. MLCK C37885A was also associated with postexercise strength loss (P < 0.05); the heterozygotes demonstrated greater strength loss compared with the homozygous wild type (CC). These results show that variations in genes coding for specific myofibrillar proteins influence phenotypic responses to muscle damaging exercise. PMID- 15817726 TI - In vivo evidence against a role for adenosine in the exercise pressor reflex in humans. AB - The pressor response to exercise is of great importance in both physiology and pathophysiology. Whether endogenous adenosine is a trigger for this reflex remains controversial. Muscle interstitial adenosine concentration can be determined by microdialysis. However, there are indications that local muscle cell damage by the microdialysis probe confounds these measurements in exercising muscle. Therefore, we used the nucleoside uptake inhibitor dipyridamole as pharmacological tool to bypass this confounding. We used microdialysis probes to measure endogenous adenosine in forearm skeletal muscle of healthy volunteers during two cycles of 15 min of intermittent isometric handgripping. During the second contraction, dipyridamole (12 microg.min(-1).dl forearm(-1)) was administered into the brachial artery. Dipyridamole potentiated the exercise induced increase in dialysate adenosine from 0.30 +/- 0.08 to 0.48 +/- 0.10 micromol/l (n = 9, P < 0.05), but it did not potentiate the exercise-induced increase in blood pressure. A time-control study without dipyridamole revealed no difference in exercise-induced increase in adenosine between both contractions (n = 8). To exclude the possibility that the dipyridamole-induced increase in dialysate adenosine originates from extravasation of increased circulating adenosine, we simultaneously measured adenosine with microdialysis probes in forearm muscle and antecubital vein. In a separate group of nine volunteers, simultaneous intrabrachial infusion of 100 microg.min(-1).dl(-1) dipyridamole and 5 microg.min(-1).dl(-1) adenosine increased dialysate adenosine from the intravenous but not the interstitial probe, indicating preserved endothelial barrier function for adenosine. We conclude that dipyridamole significantly inhibits uptake of interstitial adenosine without affecting the pressor response to exercise, suggesting that interstitial adenosine is not involved in the pressor response to rhythmic isometric exercise. PMID- 15817727 TI - Cardiac structure and function in monozygotic twin pairs discordant for physical fitness. AB - Cross-sectional studies in athletes and untrained subjects suggest that exercise training induces adaptations in cardiac structure and function. However, the role of genetic variation on the results has largely been ignored in these studies. The purpose of this study was to investigate the effects of long-term volitionally increased physical activity on electrocardiographic and echocardiographic parameters in male monozygotic twin pairs discordant for physical activity and fitness. On the basis of the mailed questionnaires, a telephone interview, and the inclusion criteria, 12 pairs of young adult male monozygotic twins were recruited from a Finnish twin cohort. All subjects completed a maximal oxygen uptake (.VO2 (max)) test and electrocardiography and echocardiography studies. Nine pairs had at least 9% difference in .VO2(max) and were selected for further analysis and for a second echocardiography study. Twins were divided into the more (MAG) and less active group (LAG), according to their VO2(max). On average, MAG had 18% higher VO2(max) compared with LAG. In electrocardiography, MAG had 29% (P = 0.02) higher Cornell voltage and 37% (P = 0.01) higher right-side hypertrophy index. In echocardiography, no significant differences were observed between the groups, and left ventricular mass index was only 7% (P = 0.16) higher in MAG. These results show that the volitionally increased physical activity that has led to an 18% increase in cardiorespiratory fitness induces greater changes in electro- than echocardiographic parameters. Electrocardiographic changes were suggestive of left ventricular hypertrophy, and echocardiography showed a similar but statistically nonsignificant trend. PMID- 15817728 TI - Why is violence more common where inequality is greater? AB - The most well-established environmental determinant of levels of violence is the scale of income differences between rich and poor. More unequal societies tend to be more violent. If this is a relation between institutional violence and personal violence, how does it work and why is most of the violence a matter of the poor attacking the poor rather than the rich? This paper begins by showing that the tendency for rates of violent crime and homicide to be higher where there is more inequality is part of a more general tendency for the quality of social relations to be poorer in more hierarchical societies. Research on the social determinants of health is used to explore these relationships. It is a powerful source of insights because health is also harmed by greater inequality. Because epidemiological research has gone some way towards identifying the nature of our sensitivity to the social environment and to social status differentials in particular, it provides important insights into why violence is related to inequality. The picture that emerges substantiates and explains the common intuition that inequality is socially corrosive. With an evolutionary slant, and informed by work on ranking systems in non-human primates, this paper focuses on the sharp distinction between competitive social strategies appropriate to dominance hierarchies and the more affiliative social strategies associated with more egalitarian social structures. The implications for policy seem to echo the importance to the quality of life of the three inter-related dimensions of the social environment expressed in the demand for "liberty, equality, fraternity." PMID- 15817729 TI - Dangerous and endangered youth: social structures and determinants of violence. AB - Structural violence is violence that is permissible, even encouraged. It refers to the invisible social machinery of inequality that reproduces social relations of exclusion and marginalization via ideologies, stigmas, and dangerous discourses (such as "youth violence" itself) attendant to race, class, sex, and other invidious distinctions. Structural violence "naturalizes" poverty, sickness, hunger, and premature death, erasing their social and political origins so that they are taken for granted and no one is held accountable except the poor themselves. Structural violence also refers to the ease with which humans are capable of reducing the socially vulnerable (even those from their own class and community) into expendable non-persons, thus allowing the licence--even the duty- to kill them. I exemplify this through two ethnographic critical case studies: the operation of a virulent death squad in Northeast Brazil that mobilized the support of ordinary people in an almost genocidal attack against Afro-Brazilian street kids and young "marginals"; and the uneasy truce with, and incomplete integration of "dangerous and endangered" youth still living in squatter camps and shack communities of urban South Africa. PMID- 15817730 TI - Social stratification, health, and violence in the very young. AB - Inequalities in human social groups contribute in important ways to the initiation and perpetuation of violence. Because both social inequalities and interpersonal aggression emerge in early life, it is important to study their developmental origins in the collective and individual behavior of young children. As young as 2 years of age, children assemble stable, linearly transitive dominance hierarchies when brought together in novel social groups. Just as the consequences of socioeconomic status may be due in part to experiences of social ordering per se, subjective childhood experiences of occupying a lesser or greater position on a scale of social influence may similarly affect health, safety, and behavior in early development. Children's experiences with social ordering may represent a first, formative encounter with the hierarchical social relations that affect health and susceptibility to violence over the human lifespan. We have studied the health correlates of group stressors, dominance positions, and biobehavioral reactivity within cohorts of preschool children and semi free-ranging rhesus macaques. Social position is ascertained using naturalistic observations of critical dyadic events, and stress reactivity is measured using assessments of behavioral and biological responses to standardized challenges. Outcome measures, including indicators of mental and physical health and interpersonal injuries, have been assessed--in the case of children--with parent- and teacher-report questionnaires, child self-report protocols, and frequent, objective physical examinations of the child and--in the case of rhesus monkeys--with daily observations of violence-related injuries. Results to date have suggested the following provisional conclusions: (1) Children and monkeys form stable, linearly transitive social hierarchies in which identifiable subgroups of individuals occupy dominant and subordinate social positions. (2) Individuals occupying subordinate positions within the social group show exaggerated adrenocortical and/or autonomic reactivity to challenge and disproportionate rates of chronic medical conditions or violent injuries. (3) Stress-reactivity, naturally occurring stressors, and social position are interactively predictive of rates of morbidity. Monkeys who displayed high biobehavioral reactivity sustained disproportionate numbers of violent injuries during a confinement stressor. Children who were high in the dominance hierarchy and low in reactivity showed significantly higher rates of externalizing mental health symptoms, while those who were in subordinate social positions and high in reactivity showed higher prevalences of internalizing symptoms. In conclusion, the prevention of youth violence--and other forms of early disorder--will require a deeper, developmental understanding of the "headwaters" of aggression and victimization in early childhood and a stronger accounting of how early social inequalities set trajectories toward healthy or disordered behavior. PMID- 15817731 TI - The discourse on violence prevention: what are the implications for smaller schools? AB - A seemingly rigorous positivistic scientific research that claims only to record the "objective facts" about school violence (e.g., by identifying "persistently dangerous schools" or recording the numbers of "violent incidents" in a particular school) in fact constructs the very subject (the violent student) that it purports to describe. This dominant discourse, reinforced by funding agencies that insist on "evidenced-based" results and randomized controlled trials (said to be the "gold standard" for establishing "what works"), have reduced the unit of analysis to the individual adolescent (who finally becomes defined as the "perpetrator") instead of investigating the societal institutions (e.g., large overcrowded schools, the prevalence of guns, violent films) which undergird the violence. This paper rejects such simplistic and reductionistic explanations for youth violence and argues that psychoanalysis, ethnography, and social interactionist disciplines reveal a much more complex picture of the contexts in which youth violence occurs. The small school movement in conjunction with the appearance of the initiative known as social and emotional education are two hopeful phenomena that have emerged to address some of these social inequities and injustices. PMID- 15817732 TI - Social cognition as a mediator of the influence of family and community violence on adolescent development: implications for intervention. AB - Several studies have shown that exposure to family and community violence during childhood and adolescence is associated with an increased risk for development of externalizing behavior problems, but less is known about specific mechanisms that mediate this relation. Variations in social cognition serve as one possible mechanism by which these environmental experiences influence aggressive behavior during adolescence. Children who have been maltreated tend to display negatively biased social-cognitive processing styles, which may in turn increase their likelihood of reacting aggressively in ambiguous social situations. Similarly, witnessing community violence is associated with aspects of social cognition, including beliefs that support aggressive responses to threat. Recent studies also suggest that exposure to extreme forms of stress and violence can produce changes in children's neurobiology that may increase their hypersensitivity and reactivity to interpersonal threat. Some of the strongest evidence of the role of social cognition as a mechanism in this association comes from intervention studies which reduced aggressive behavior by targeting negatively biased social cognitive processing styles. PMID- 15817733 TI - Biological aspects of social bonding and the roots of human violence. AB - The brain systems that motivate humans to form emotional bonds with others probably first evolved to mobilize the high-quality maternal care necessary for reproductive success in placental mammals. In these species, the helplessness of infants at birth and their dependence upon nutrition secreted from their mothers' bodies (milk) and parental body heat to stay warm required the evolution of a new motivational system in the brain to stimulate avid and sustained mothering behavior. Other types of social bonds that emerged subsequently in placental mammals, in particular monogamous bonds between breeding pairs, appear to have evolved from motivational brain systems that stimulate maternal behavior. This chapter focuses on aspects of the evolution and neurobiology of maternal and pair bonding and associated behavioral changes that may provide insights into the origins of human violence. The roles of the neuropeptides oxytocin and vasopressin as well as the neurotransmitter dopamine will be emphasized. Maternal and pair bonding are accompanied by increased aggressiveness toward perceived threats to the object of attachment as well as diminished fear and anxiety in stressful situations. The sustained closeness with mother required for the survival of infant mammals opened a new evolutionary niche in which aspects of the mother's care became increasingly important in regulating development in offspring. The quantity and quality of maternal care received during infancy determines adult social competence, ability to cope with stress, aggressiveness, and even preference for addictive substances. Indeed, the development of neurochemical systems within the brain that regulate mothering, aggression, and other types of social behavior, such as the oxytocin and vasopressin systems, are strongly affected by parental nurturing received during infancy. Evidence will be reviewed that the neural circuitry and neurochemistry implicated in studies of lower mammals also facilitate primate/human interpersonal bonding. It is hypothesized that neural bonding systems may also be important for the development in individuals of loyalty to the social group and its culture. Neglect and abuse during early life may cause bonding systems to develop abnormally and compromise capacity for rewarding interpersonal relationships and commitment to societal and cultural values later in life. Other means of stimulating reward pathways in the brain, such as drugs, sex, aggression, and intimidating others, could become relatively more attractive and less constrained by concern about violating trusting relationships. The ability to modify behavior based on negative experiences may be impaired. Unmet needs for social bonding and acceptance early in life might increase the emotional allure of groups (gangs, sects) with violent and authoritarian values and leadership. Social neurobiology has the potential to provide new strategies for treating and preventing violence and associated social dysfunction. PMID- 15817734 TI - Strengthening healthy resistance and courage in children: a gender-based strategy for preventing youth violence. AB - Beginning with the distinction between core consciousness or a core sense of self and a self that is wedded to a story about itself, this paper suggests that we have collectively been wedded to a false story about ourselves, a story that the core self resists. The gender disparity with respect to times in development when children's resilience is at heightened risk highlights the costs of an initiation that occurs for boys in early childhood and for girls at adolescence. Because of this difference in the timing, girls can become informants about a process of psychic splitting and dissociation that impedes the relational capacities of children and opens the way to violence. The articulateness of girls' resistance to losses that are psychologically and socially consequential illuminates a resistance in boys that may otherwise be overlooked. Evidence drawn from studies of girls' development leads to the suggestion that by joining a healthy resistance in children, we can act to prevent youth violence. The paper ends with a case study of a suicidal adolescent girl to illustrate how a relational framework shifts the interpretation of violent behavior and informs a strategy of response. PMID- 15817735 TI - Male adolescent rites of passage: positive visions of multiple developmental pathways. AB - Unlike the separation-based, stereotyped views of boys' developmental movement into adulthood, this paper will argue that there are more modern and relational models, as well as multiple pathways, for young males to journey through such rites of passage. Indeed, it will be suggested and supported by both qualitative and quantitative data that the more classic models depend on a "boy code" of traumatic separation from mother and the feminine, a process that is not only negative rather than positive in its developmental trajectory, but also likely to create a premature traumatic separation, leaving boys at risk for emotional maladjustment, everyday sadness, increased incidence of depression and the potential for violence toward the self, suicide, as well as violence toward others. More-positive visions and versions of male rites of passage will be posited and described. The definition of emotional "resilience" during this significant period will be re-addressed as one of "healthy vulnerability," sustained through connection to loving adults, rather than a classic belief in stoicism and release from relational ties. Attachment theory will be brought to bear and the desperate yearnings of adolescent males not only for connection to adult mentors, but also for non-romanticized friendships with adolescent females, will be discussed. Finally, the understanding and substitution of these new, more positive, developmental pathways will be linked to the prevention of violence. PMID- 15817736 TI - Age, puberty, and exposure to intimate partner violence in adolescence. AB - This paper links sociological and epidemiologic research on violence and the life course to biosocial perspectives on pubertal maturation to examine risk factors associated with exposure to intimate partner violence in adolescence. While prior research has established early puberty as a risk factor for delinquent behavior, studies to date have not yet investigated whether early puberty is also linked to intimate partner violence in adolescence. Prior epidemiologic research has found that increasing age in adolescence is a risk factor for dating violence, but this work has not yet incorporated the element of pubertal maturation. The present study examines the relative effects of chronological age and maturational age in a biosocial model predicting risk for intimate partner violence among adolescent females, net of established control variables, using three waves of data from the National Longitudinal Study of Adolescent Health. These findings indicate that early maturation in females is an additional risk factor for exposure to intimate partner violence in adolescence. The importance of disentangling types of age effects as raised in the developmental literature and as supported by these findings is discussed in relation to the prevention of youth violence. PMID- 15817737 TI - Epigenetic programming of stress responses through variations in maternal care. AB - Early life experiences shape an individual's physical and mental health across the lifespan. Not surprisingly, an upbringing that is associated with adversity can produce detrimental effects on health. A central theme that arises from studies in human and nonhuman species is that the effects of adversity are mediated by the interactions between a mother and her young. In this review we describe some of the long-term effects of maternal care on the offspring and we focus on the impact of naturally occurring variations in the behavior of female rats. Of particular interest are mothers that engage in high or low amounts of licking/grooming (LG) and arched-back nursing (ABN) of their pups, but do so within the normal range for this species. Such variations in LG-ABN can alter the function of the hypothalamic-pituitary-adrenal (HPA) axis, and cognitive and emotional development by directly affecting the underlying neural mechanisms. At the heart of these mechanisms is gene expression. By studying the hippocampal glucocorticoid receptor gene, we have identified that maternal care regulates its expression by changing two processes: the acetylation of histones H3-K9, and the methylation of the NGFI-A consensus sequence on the exon 1(7) promoter. Sustained "maternal effects" appear elsewhere in biology, including plants, insects, and lizards, and may have evolved to program advantages in the environments that the offspring will likely face as adults. Given the importance of early life and parent-child interactions to later behavior, prevention and intervention programs should target this critical phase of development. PMID- 15817738 TI - Early-life trauma and the psychogenesis and prevention of violence. AB - This article considers the development of violence with particular reference to family factors in violence such as the quality of the parent-child relationship. In taking a developmental approach to violence, a link is established between the maltreatment of children in an attachment context and the risk of violence via the child's capacity to envision mental states in the other. Evidence from epidemiology and neuroscience is brought to bear on this link. Finally, some studies of prevention of violence that are likely to enhance attachment and mentalizing are considered. PMID- 15817739 TI - Cost-effective violence prevention through targeted family interventions. AB - Efforts in violence prevention can focus on individual youth, their families, their schools, or the communities in which they live. Among the small number of program models that have been proven effective in repeated replications, those that focus on improving family management and child care have been found to be the most cost-effective. The four model programs include Nurse Home Visitation, Functional Family Therapy, Multi-systemic Therapy, and Multidimensional Foster Care. All of these programs involve detailed protocols, extensive staff training and supervision, and quality-assurance procedures. The factors limiting their wider adoption include staff resistance to their structured approach, cost sharing issues between local and state levels of government, and the political power of existing programs. PMID- 15817740 TI - The role of the bystander in the social architecture of bullying and violence in schools and communities. AB - The bystander is defined as an active and involved participant in the social architecture of school violence, rather than a passive witness. Bullying is redefined from a triadic (bully-victim-bystander) rather than dyadic (bully victim) perspective. Teachers, including administrators, and students can promote or ameliorate bullying and other forms of violence when in this social role. Cases are used to illustrate this phenomenon, including one in which a teacher is murdered. Data are presented from a study of teachers' perceptions of other teachers who bully students, suggesting that bullying of students by teachers and bullying of teachers by students is a factor in the aggravation of school bullying and violence that needs to be more openly discussed. An intervention in nine elementary schools involving 3,600 students is outlined to illustrate how a focus on reflective mentalizing and awareness of the importance of the helpful bystander role can promote a peaceful school-learning environment for students and teachers. The paper concludes with an outline for research into how communities and schools adopt bystanding roles when faced with complex problems like youth violence, and how they may avoid facing the problems by blaming law enforcement and educators. PMID- 15817741 TI - Collective violence: comparisons between youths and chimpanzees. AB - Patterns of collective violence found among humans include similarities to those seen among chimpanzees. These include participation predominantly by males, an intense personal and group concern with status, variable subgroup composition, defense of group integrity, inter-group fights that include surprise attacks, and a tendency to avoid mass confrontation. Compared to chimpanzee communities, youth gangs tend to be larger, composed of younger individuals, occupying smaller territories and having a more complex organization. Youth gangs also differ from chimpanzee communities as a result of numerous cultural and environmental influences including complex relations with non-gang society. These relations are governed in important ways by such factors as perceived economic and personal constraints, policing, family structure, and levels of poverty, crime, and racism. Nevertheless, the concepts that sociologists use to account for collective violence in youth gangs are somewhat similar to those applied by anthropologists and biologists to chimpanzees. Thus in both cases collective violence is considered to emerge partly because males are highly motivated to gain personal status, which they do by physical violence. In the case of youth gangs, the reasons for the prevalence of physical violence in status competition compared to non-gang society are clearly context-specific, both culturally and historically. By contrast, among chimpanzees the use of physical violence to settle status competition is universal (in the wild and captivity). The use of physical violence in individual status competition therefore has different sources in youth gangs and chimpanzees. Regardless of its origin, however, its combination with an intense concern for status can explain: (1) why individual males form alliances among each other; and hence (2) how such alliances generate social power, closed groups, and a capacity for defense of territory or pre emptive attacks on rivals. This comparison suggests that the use of physical violence to resolve individual status competition is an important predictor of collective violence at the gang level. We therefore view the similarities in aggression between humans and chimpanzees that we review here as being adaptive responses to local conditions, predicated on an inherent male concern for social status. PMID- 15817742 TI - Genetic influences on aggressive behaviors and arousability in animals. AB - In a variety of species, strain differences in aggressive behaviors strongly indicate genetic influences. In people, as suggested, for example, by the work of Kagan, Rothbart, Cloninger, and their collaborators, long-lasting differences in temperament argue for genetic as well as environmental effects. After well controlled experimentation in mice, we have learned five lessons about gene/behavior causal relations bearing on sociosexual and aggressive behaviors. The effect of a given gene on a given behavior depends upon: (1) exactly when and where that gene is expressed in the brain; (2) the gender of the animal in which it is expressed; (3) the age of the animal; (4) the nature of the opponent; and (5) the form of aggression (e.g., testosterone-facilitated aggression vs. maternal aggression). Moreover, in female mice, better social recognition is correlated with lower levels of aggression. We have gathered evidence for a four gene micronet involving estrogen receptors alpha and beta, oxytocin, and the oxytocin receptor as expressed in the hypothalamus and amygdala. Normal performance of this micronet is required for social recognition and thus for control over aggression. Underlying certain genetic influences on sociosexual behaviors and aggression may be alterations in generalized brain arousal. PMID- 15817743 TI - On the nature and nurture of antisocial behavior and violence. AB - This article focuses on the contribution that behavioral genetic research can make to further the understanding of how antisocial and violent behavior develops. Genetically informative study designs are particularly useful for investigating etiological heterogeneity and can refine the search for developmental pathways to persistent antisocial conduct. While the current data are not yet directly translatable for prevention programs, behavioral genetic research will have far-reaching implications for prevention and treatment. As we find genes associated with risk for antisocial behavior and develop better understanding of the mechanics of the interplay between genes and the environment, we can expect to tailor prevention and treatment to serve the specific needs of etiologically distinct subgroups of children. Furthermore, researchers involved in sharpening the knowledge base have the responsibility of trying to ensure that such findings are not misused. PMID- 15817744 TI - Role of alcohol consumption in escalation to violence. AB - No other drug has been associated with aggressive and violent behavior more than alcohol has. A major characteristic of the link between alcohol and social interactions is the very large variation in who becomes more aggressive while drinking and who does not. Tracing the origins of these individual differences has led to a focus on predispositions, such as the antisocial behavior of Type 2 alcoholics. Successful development of an experimental procedure to model heightened aggressive behavior after voluntary consumption of alcohol has facilitated the neurobiologic analysis of the link between alcohol and aggression. From a pharmacologic perspective, consumption of low to moderate doses of alcohol engenders heightened aggressive behavior in a significant minority of individuals before the circulation of appreciable amounts of the aldehyde metabolite. Ionophoric receptors such as NMDA, 5-HT(3) and GABA(A) have been identified in the brain as major sites of action for alcohol in the dose range that is relevant for engendering heightened aggression. Actions at the GABA(A) receptor complex that depend on particular GABA(A) subunits appear to be necessary for alcohol-heightened aggression. Genes that encode the synthesis of these alpha and gamma subunits are potentially significant markers for those individuals that are prone to engage in heightened aggressive behavior after the consumption of alcohol. Of particular importance are the reciprocal interactions between GABA and serotonin. Activating specific serotonin receptor subtypes such as 5-HT(1B) receptors reduces alcohol-heightened aggressive behavior. How these GABAergic and serotonergic corticolimbic mechanisms for alcohol-heightened aggression develop during the adolescent period remains an area of urgent study. PMID- 15817745 TI - The discrete character of high-lethality youth violence. AB - The purpose of this paper is to derive lessons about high-lethality adolescent violence from study of the extraordinary increase, and then decrease, in youth homicide arrests over the period 1985-2001 in the United States. This effort continues a long tradition of exploring the possible causes of shifts in homicide rates by examining patterns of violence to see whether there are important changes in types of violence. The first section of the paper discusses two dimensions of criminal youth violence over the whole range of reported offenses. The second section shows that one subset of attacks--firearms cases--accounts for the major changes in youth homicide when it increased from 1985-1993 and during the years of decrease, 1994-2001. A third section explores the linkage between rates of adolescent homicide in various cultures and the relative rate of homicide among all ages. My central conclusion is that high-lethality violence among youth is not representative of the fighting and group assaults that are relatively frequent among adolescents. Instead, the attacks that often lead to death differ with respect to the use of weapons and, to some extent, geography. But there is usually a strong relationship between homicide rates for all ages and youth homicide rates in a particular location. PMID- 15817746 TI - Beyond the prison paradigm: from provoking violence to preventing it by creating "anti-prisons" (residential colleges and therapeutic communities). AB - Prisons were supposedly created for the purpose of the tertiary prevention of violence (i.e., reducing the frequency and severity of future violence on the part of people who have already become violent). However, there is much evidence that this method of attempting to prevent violence is often, though not always, either ineffectual or counterproductive, in which case it is either a waste of money or actually exacerbates the problem it was ostensibly intended to solve. This article reviews evidence concerning those questions including an analysis of the effect of punishment (one of the main purposes of prisons) on violent behavior. Punishment--the infliction of pain--will be distinguished from restraint (incapacitation, separation from the community). Successful examples of violence prevention in unconventional prison programs, emphasizing therapy and education rather than punishment, and restorative rather than retributive justice, will be summarized, together with evidence that these programs reduce re incarceration rates so substantially that they actually save the taxpayers more money than they cost, in addition to enhancing the safety of the general public. The position is taken that traditional prisons provoke more violence than they prevent and are so fundamentally flawed that they cannot be reformed; we argue that they should instead be abolished and replaced by "anti-prisons," that is, locked, secure residential colleges, therapeutic communities, and centers for human development. Prisons will come to be seen as a well-meaning experiment that failed, rather like the use of leeches in medicine. PMID- 15817747 TI - The behavioral economics of violence. AB - From the viewpoint of teleological behaviorism the first question to ask in attempting to understand any behavior, including violent behavior, is: What are its contingencies of reward and punishment? Or, to put the question in economic terms: What are the short-term and long-term costs and benefits that such behavior entails? Let us therefore consider the costs and benefits of youth violence. Among the short-term costs of violent behavior are the physical effort of the act, the possibility of immediate physical retaliation, immediate social disapproval, and the opportunity cost of other social acts that the violent behavior takes the place of (you can't be affectionate and violent at the same time, for instance). Among the immediate benefits of violent behavior are the intrinsic satisfaction of the violent act itself and any extrinsic benefit; if A violently appropriates B's new sneakers then obtaining the sneakers reinforces A's violence. These immediate benefits may well outweigh the costs in many contexts. Among the long-term costs of violent behavior are delayed retaliation, possible social disapproval and loss of social support, rejection from a social group, job loss, and health risks associated with a violent lifestyle. Among the long-term benefits are long-term intimidation of others (your neighbor is less likely to build a fence on your property if you have a reputation for violence), and a possibly exciting lifestyle. These long-term benefits may well be outweighed by the long-term costs. Opposition of long-term net costs to short term net benefits, where it exists, creates a personal self-control trap: Overall satisfaction may decrease monotonically with rate of the target behavior but, regardless of its rate, the immediate satisfaction of doing it is always higher than that of not doing it. In the case of violent behavior, this trap is exacerbated by the fact that as a person's violence increases, net immediate reinforcement also increases (due to membership in violence-reinforcing subgroups). This contingency fits the "primrose path" addiction model of Prelec and Herrnstein. Violence is thus a paradigm case of behavioral addiction. I consider three ways of controlling such addictive behavior: by punishment, by extinction, and by substitution. The problem with punishment in the case of violence is that physical punishment tends to increase violent behavior while incarceration drives the punished person into the very social subgroup (the prison culture) where violence is maximally reinforced. The problem with extinction is that the immediate benefits of violent behavior are largely intrinsic and some costs (immediate retaliation by unidentified others) are difficult to control. The best way to control violent behavior, as well as other addictive behaviors, is by decreasing the price of economic substitutes. There is much evidence that addictions, such as to cocaine, heroin, alcohol, and tobacco, may be reduced by decreasing the price of social support. The same is predicted for violent behavior--either by providing social support directly or by training in social skills. In addition, in considering control of violent behavior, we need to examine the immediate benefits and long-term costs to society of having violent individuals and violence-reinforcing subcultures among us. And we need to act to reduce our own dependence on those benefits. PMID- 15817748 TI - Escalated aggressive behavior: new pharmacotherapeutic approaches and opportunities. AB - Psychopharmacologic studies of aggressive behavior in animals under controlled laboratory conditions have been instrumental in developing and evaluating specific and effective novel drug treatments that reduce aggressive behavior. An initial contribution of this research is to create experimental conditions that enable the display of aggressive and defensive acts and postures in species that engage in either dominance or territorial or maternal aggression. Quantitative ethological analyses allow the precise delineation of the sequential organization of aggressive bursts, providing a benchmark for assessing excessive or pathological forms of aggressive behavior. A second contribution of preclinical research is the development of experimental models of escalated forms of aggressive behavior, such as focusing on genetic predispositions or social provocations and frustrative experiences. A critical role of preclinical research is in the pharmacological and neurochemical analysis of aggressive behavior; for example, a host of undesirable side effects prompted a shift from classic dopaminergic neuroleptic compounds to the more recently developed atypical neuroleptics with effective and more specific anti-aggressive effects. The long established role of brain serotonin in impulsive and escalated forms of aggressive behavior continues to be a focus of preclinical studies. New evidence differentiates dynamic state changes in corticolimbic serotonergic neurons during the termination of aggressive behavior from the deficient-serotonin trait in violence-prone individuals. It can be anticipated that currently developed tools for targeting the genes that code for specific subtypes of serotonin receptors will offer new therapeutic options for reducing aggressive behavior, and the 5 HT(1B) receptor appears to be a promising target. The modulation of GABA and GABA(A) receptors by 5-HT in corticolimbic neurons promises to be particularly relevant for specific forms of escalated aggressive behavior such as alcohol heightened aggression. PMID- 15817749 TI - The psychopharmacologic treatment of violent youth. AB - Aggressive violence has been described as the greatest problem and the most frequent reason for referrals in child and adolescent psychiatry. In this country we have only partially emerged from an epidemic of violence that was really an epidemic of youth violence. Thus it is hardly surprising that psychiatrists are being asked more and more frequently whether psychiatric medications might help to diminish the toll from this behavioral plague. Medications are useful and appropriate for only a small minority of the people who commit serious violence. Even when they are indicated, they can never be the sole treatment modality, but should be supplemented by psychological and social therapies. When the violence is a byproduct or symptom of an underlying mental illness, treating that illness is generally the most effective method of preventing future violence on a long term basis. However, most violence is not committed by those who are mentally ill, and most of the mentally ill never commit a serious act of violence. That is why many attempts have been made to discover whether there are drugs that diminish the symptom, violence, even when there is no underlying mental illness for which drugs would normally be prescribed. In fact there are several, and their indications and use are reviewed here. Different principles govern the acute short-term emergency treatment of a violent crisis and the long-term treatment of those who are chronically and repetitively violent, and these differences are also summarized here. PMID- 15817750 TI - Serotonin and aggression. AB - The neurotransmitter serotonin (5-HT) has been implicated in the modulation of aggression in animals and humans. A longstanding dogma that aggression and serotonergic activity are inversely related has to be abandoned in light of many new findings. Trait and state aggression are differentially regulated by the 5-HT system and different 5-HT receptors seem to be involved. Of the 14 different 5-HT receptors, the 5-HT(1B) receptor, particularly the postsynaptically located 5 HT(1B) heteroreceptor, plays a highly selective role in the modulation of offensive aggression. We are still far from understanding the complex role played by the serotonergic system in the modulation of a complex set of behaviors like aggression. PMID- 15817751 TI - Catecholamines and aggression: the role of COMT and MAO polymorphisms. AB - Catecholaminergic systems are involved in the regulation of aggressive behavior; this regulation is implemented in interactions with other neurobiological mechanisms. Most of the available evidence indicates that norepinephrine and dopamine lower the threshold for an aggressive response to environmental stimuli. Two major enzymes are responsible for catecholamine catabolism in the brain: catechol-O-methyltransferase (COMT) and monoamine oxidase A (MAOA). The transcriptional activity of the genes coding for these enzymes is governed by common functional polymorphisms. If aggressive behavior is enhanced by catecholaminergic activity, then the lower activity of COMT and MAOA (resulting in a slower inactivation of catecholamines) should indirectly enhance aggression. This prediction has been supported by most (but not all) observations in rodents and humans. Male mice that have either the COMT or the MAOA gene knocked out show elevated aggression. The allele that codes for the lower enzymatic activity of COMT has been associated with elevated aggressive behavior in several samples of psychiatric patients. Similarly, the alleles that code for the lower activity of MAOA were associated with the development of aggressive behavior in maltreated male children in a large birth cohort study. Collectively, these results suggest that COMT and MAOA polymorphisms represent a basic neurobiological mechanism that contributes to the regulation of aggressive behavior. PMID- 15817752 TI - Anabolic androgenic steroids and aggression: studies using animal models. AB - The use of anabolic androgenic steroids (AASs) has escalated in teenagers and is associated with increased violence. Adolescent exposure to chronic high levels of AASs is of particular concern because puberty is a hormonally sensitive period during which neural circuitry for adult male patterns of behavior develop. Thus, teenage AAS use may have long-term repercussions on the potential for displaying aggression and violence. Animal models have contributed valuable information on the effects of AAS use. For example, studies in rodents confirmed that exposure to the AASs testosterone and nandrolone, but not stanozolol, does indeed increase aggression. A side effect of AAS use reported in humans is "'roid rage," characterized by indiscriminate and unprovoked aggression. Results of animal studies demonstrated that pubertal rats receiving AASs respond appropriately to social cues as they are more aggressive toward intact males than are castrates. Also, testosterone-treated males recognize appropriate environmental cues as they are most aggressive in their home cage. Thus, adolescent AAS exposure increases aggressive behaviors, but does not induce indiscriminate aggression. To assess whether AAS exposure increases aggression after provocation, rats were tested following a mild tail-pinch. In adolescent males, provocation increased aggression after withdrawal from testosterone, nandrolone, and stanozolol, an effect which persisted for many weeks. The data suggest that AASs sensitize animals to their surroundings and lower the threshold to respond to provocation with aggression. Thus, in humans, pubertal AAS exposure may not cause violent behaviors, but may increase the likelihood that aggressive acts will result in violence. This may persist into adulthood. PMID- 15817753 TI - Simplified fluorescent multiplex PCR method for evaluation of the T-cell receptor V beta-chain repertoire. AB - RATIONALE: Evaluation of the T-cell receptor (TCR) V beta-chain repertoire by PCR based CDR3 length analysis allows fine resolution of the usage of the TCR V beta repertoire and is a sensitive tool to monitor changes in the T-cell compartment. A multiplex PCR method employing 24 labeled upstream V beta primers instead of the conventionally labeled downstream C beta primer is described. METHOD: RNA was isolated from purified CD4 and CD8 T-cell subsets from umbilical cord blood and clinical samples using TRI reagent followed by reverse transcription using a C beta primer and an Omniscript RT kit. The 24 V beta primers were multiplexed based on compatibility and product sizes into seven reactions. cDNA was amplified using 24 V beta primers (labeled with tetrachloro-6-cardoxyfluorescein, 6 carboxyfluorescein, and hexachloro-6-carboxyfluorescein), an unlabeled C beta primer, and Taqgold polymerase. The fluorescent PCR products were resolved on an automated DNA sequencer and analyzed using the Genotyper 2.1 software. RESULTS: V beta spectratypes of excellent resolution were obtained with RNA amounts of 250 ng using the labeled V beta primers. The resolution was superior to that obtained with the labeled C beta primer assay. Also the numbers of PCRs were reduced to 7 from the 12 required in the C beta labeling method, and the sample processing time was reduced by half. CONCLUSION: The method described for T-cell receptor V beta-chain repertoire analysis eliminates tedious dilutions and results in superior resolution with small amounts of RNA. The fast throughput makes this method suitable for automation and offers the feasibility to perform TCR V beta repertoire analyses in clinical trials. PMID- 15817754 TI - Identification of the gene encoding a 38-kilodalton immunogenic and protective antigen of Streptococcus suis. AB - In our continued effort to search for a Streptococcus suis protein(s) that can serve as a vaccine candidate or a diagnostic reagent, we constructed and screened a gene library with a polyclonal antibody raised against the whole-cell protein of S. suis type 2. A clone that reacted with the antibody was identified and characterized. Analysis revealed that the gene encoding the protein is localized within a 2.0-kbp EcoRI DNA fragment. The nucleotide sequence contained an open reading frame that encoded a polypeptide of 445 amino acid residues with a calculated molecular mass of 46.4 kDa. By in vitro protein synthesis and Western blot experiments, the protein exhibited an electrophoretic mobility of approximately 38 kDa. At the amino acid level the deduced primary sequence shared homology with sequences of unknown function from Streptococcus pneumoniae (89%), Streptococcus mutans (86%), Lactococcus lactis (80%), Listeria monocytogenes (74%), and Clostridium perfringens (64%). Except for strains of serotypes 20, 26, 32, and 33, Southern hybridization analysis revealed the presence of the gene in strains of other S. suis serotypes and demonstrated restriction fragment length differences caused by a point mutation in the EcoRI recognition sequence. We confirmed expression of the 38-kDa protein in the hybridization-positive isolates using specific antiserum against the purified protein. The recombinant protein was reactive with serum from pigs experimentally infected with virulent strains of S. suis type 2, suggesting that the protein is immunogenic and may serve as an antigen of diagnostic importance for the detection of most S. suis infections. Pigs immunized with the recombinant 38-kDa protein mounted antibody responses to the protein and were completely protected against challenge with a strain of a homologous serotype, the wild-type virulent strain of S. suis type 2, suggesting that it may be a good candidate for the development of a vaccine that can be used as protection against S. suis infection. Analysis of the cellular fractions of the bacterium by Western blotting revealed that the protein was present in the surface and cell wall extracts. The functional role of the protein with respect to pathogenesis and whether antibodies against the antigen confer protective immunity against diseases caused by strains of other pathogenic S. suis capsular types remains to be determined. PMID- 15817755 TI - Prospective evaluation of a whole-blood test using Mycobacterium tuberculosis specific antigens ESAT-6 and CFP-10 for diagnosis of active tuberculosis. AB - A new immunodiagnostic test based on the Mycobacterium tuberculosis-specific antigens CFP-10/ESAT-6(QFT-RD1) has been launched as an aid in the diagnosis of latent tuberculosis (TB) infection (LTBI). The aim of this study was to evaluate this test for the diagnosis of active TB. Eighty-two patients with suspicion of TB and 39 healthy BCG-vaccinated persons were enrolled. Forty-eight had active TB, 25 did not, and 9 were excluded. Sensitivity and specificity of the test for active TB were evaluated in a prospective blinded manner in patients suspected of TB. The sensitivity of the QFT-RD1 was 85% (40/48; confidence interval [CI], 75 to 96), and it was higher than the sensitivity of microscopy, 42% (20/48; CI, 27 to 56; P = 0.001), and culture, 59% (27/46; CI, 44 to 73; P = 0.009). Of patients with extrapulmonary TB, 92% (12/13) were QFT-RD1 positive, whereas only 31% (4/13) were positive by microscopy and 42% (5/12) by culture (P < 0.05), and 87% (13/15) of those who were negative by both microscopy and culture were QFT-RD1 positive. By combining microscopy and culture with the QFT-RD1 test, sensitivity increased to 96% (CI, 90 to 102). Ten of 25 (40%) non-TB patients were QFT-RD1 positive, resulting in a specificity of 60%. However, 80% (8/10) of these had risk-factors for TB, indicating latent infection in this group. In healthy controls, only 3% (1/39) were QFT-RD1 positive. In conclusion, the QFT-RD1 test is sensitive for diagnosis of TB, especially in patients with negative microscopy and culture. The accuracy of the QFT-RD1 test will vary with the prevalence of LTBI. We suggest that the QFT-RD1 test could be a very useful supplementary tool for the diagnosis of TB. PMID- 15817756 TI - Korean red ginseng slows depletion of CD4 T cells in human immunodeficiency virus type 1-infected patients. AB - We have previously showed that long-term intake of Korean red ginseng (KRG) delayed disease progression in human immunodeficiency virus type 1 (HIV-1) infected patients. In the present study, to investigate whether this slow progression was affected by KRG intake alone or in combination with HLA factor, we analyzed clinical data in 68 HIV-1-infected patients who lived for more than 5 years without antiretroviral therapy. The average KRG intake over 111.9 +/- 31.3 months was 4,082 +/- 3,928 g, and annual decrease in CD4 T cells was 35.0 +/- 28.7/microl. Data analysis showed that there are significant inverse correlations between the HLA prognostic score (0.29 +/- 1.19) and annual decrease in CD4 T cells (r = -0.347; P < 0.01) as well as between the amount of KRG intake and annual decrease in CD4 T cells (r = -0.379; P < 0.01). In addition, KRG intake significantly slowed the decrease in CD4 T cells even when influence of HLA class I was statistically eliminated (repeated-measure analysis of variance; P < 0.05). We also observed significant correlation between KRG intake and a decrease in serum-soluble CD8 antigen level (r = 0.62; P < 0.001). In conclusion, these data show that KRG intake independently and significantly affected the slow depletion of CD4 T cells irrespective of HLA class I. PMID- 15817757 TI - Assessment by flow cytometry of cytokine production in malnourished children. AB - Malnutrition in children is associated with an increased risk of infection and death. Multiple abnormalities in the immune response, including cytokine production, in protein energy-malnourished children have been described and could account for the increased severity and frequency of infections. In this study, we used flow cytometry to investigate the effects of malnutrition on the production of cytokines (interleukin-2 [IL-2], gamma interferon [IFN-gamma], IL-4, and IL 10) in CD4+ and CD8+ cells and the activation capability (as indicated by CD69+ and CD25+ cells). CD4+ and CD8+ cells from malnourished children showed increased production of IL-4 and IL-10 cytokines and decreased production of IL-2 and IFN gamma cytokines compared to that in cells from well-nourished, uninfected and well-nourished, infected children. In addition, malnourished children showed impaired activation capability, since the fluorescence intensity of CD69+ and CD25+ cells was lower than that in cells from well-nourished, uninfected and well nourished, infected children. These results indicate that malnutrition alters the capacity of CD4+ and CD8+ cells to produce IL-2, IFN-gamma, IL-4, and IL-10 in response to stimulus. We concluded that both cytokine production and activation capacity were impaired in malnourished children. This functional impairment may be involved in the failure to develop a specific immune response and the predisposition to infection in these children. PMID- 15817758 TI - Point seroprevalence survey of Ehrlichia ruminantium infection in small ruminants in The Gambia. AB - Using the MAP1-B enzyme-linked immunosorbent assay, we tested 1,318 serum samples collected from sheep and goats at 28 sites in the five divisions of The Gambia to determine the Ehrlichia ruminantium seroprevalence rates and to assess the risk for heartwater. About half (51.6%) of 639 sheep were positive, with seroprevalence rates per site varying between 6.9% and 100%. The highest seroprevalence was detected in the western part of the country (88.1% in the Western Division and 62.1% in the Lower River Division). Sheep in the two easterly divisions (Central River and Upper River divisions) showed the lowest seroprevalence of 29.3% and 32.4%, respectively, while those in the North Bank Division showed an intermediate prevalence of 40.6%. In goats, less than one third (30.3%) of 679 animals tested were positive. The highest seroprevalence was detected in goats in the North Bank Division (59%) and Western Division (44.1%). Goats in the Lower River Division showed an intermediate level of 21.9%, whereas the lowest rates were found in the eastern part of the country (4.8% in the Central River Division and 2.3% in the Upper River Division). At nearly all sites, seroprevalence rates were higher in sheep than in goats. The results show a gradient of increasing heartwater risk for susceptible small ruminants from the east to the west of The Gambia. These findings need to be taken into consideration when future livestock-upgrading programs are implemented. PMID- 15817759 TI - Simultaneous serodetection of 10 highly prevalent mouse infectious pathogens in a single reaction by multiplex analysis. AB - Under current practices of mouse colony maintenance, sera from mice are analyzed for antibodies against several widespread infectious pathogens by conventional immunoassays, generally enzyme-linked immunosorbent assay (ELISA). To test for multiple agents, these methods consume large volumes of mouse serum and are laborious and time-consuming. More efficient immunoassays, using small amounts of sample, are therefore needed. Accordingly, we have developed a novel multiplex diagnostic system that employs fluorescent microbeads, coated with purified antigens, for simultaneous serodetection of 10 mouse infectious agents. Individually identifiable, fluorescent microbeads were coated with antigens from Sendai virus, mouse hepatitis virus, Theiler's mouse encephalomyelitis virus/GDVII strain, mouse minute virus, mouse cytomegalovirus, respiratory enteric orphan virus (Reo-3 virus), mouse parvovirus, calf rotavirus for epizootic diarrhea virus of infant mice, vaccinia virus for ectromelia virus, and Mycoplasma pulmonis. Standard sera, singly positive for antibodies to individual infectious agents, were generated by inoculation of BALB/cj and C57BL/6j mice. Sera from these experimentally infected mice, as well as sera from naturally infected mice, were analyzed using a mixture of microbeads coated with antigens of the 10 infectious agents listed above. Results demonstrated that the multiplex assay was at least as sensitive and specific as ELISA for serodetection. Importantly, the multiplex assay required only 1 microliter of serum for simultaneous serodetection of the 10 mouse infectious agents in one reaction vessel. Thus, this multiplex microbead assay is a reliable, efficient, and cost effective diagnostic modality that will impact serosurveillance of mice used in research. PMID- 15817760 TI - Persisting humoral antiviral immunity within the Japanese population after the discontinuation in 1976 of routine smallpox vaccinations. AB - Concerns have arisen recently about the possible use of smallpox for a bioterrorism attack. Routine smallpox vaccination was discontinued in Japan in 1976; however, it is uncertain exactly how long vaccination-induced immunity lasts. We sought to evaluate the seroprevalence and intensity of anti-smallpox immunity among representatives of the present Japanese population. The subjects included 876 individuals who were born between 1937 and 1982. Vaccinia virus specific immunoglobulin G (IgG) levels were measured by enzyme-linked immunosorbent assay (ELISA), and 152 of 876 samples were also tested for the presence of neutralizing antibodies. Of the subjects who were born before 1962, between 1962 and 1968, and between 1969 and 1975, 98.6, 98.6, and 66.0%, respectively, still retained the vaccinia virus-specific IgG with ELISA values for optical density at 405 nm (OD(405)) of > or = 0.10. The corresponding figures for retained IgGs with OD405 values of > or = 0.30 were 91.0, 90.3, and 58.2%, respectively. Neutralizing antibodies were also maintained. The sera with OD(405) values of > or = 0.30 showed 89% sensitivity and a 93% positive predictive value for detection of neutralizing antibodies (> or = 4). Thus, approximately 80% of persons born before 1969 and 50% of those born between 1969 and 1975 were also found to have maintained neutralizing antibodies against smallpox. A considerable proportion of the previous vaccinated individuals still retain significant levels of antiviral immunity. This long-lasting immunity may provide some protective benefits in the case of reemergence of smallpox, and the disease may not spread as widely and fatally as generally expected. PMID- 15817761 TI - Reference values of CD4 T lymphocytes in human immunodeficiency virus-negative adult Nigerians. AB - A cross-sectional study that involved secondary analysis of data collected from 681 pregnant women and 183 miners (94 men and 89 women; ratio of men to women, 1:0.95) in Jos, Nigeria, was carried out to determine the reference ranges for CD4(+)-cell counts in healthy HIV-negative adult Nigerians. The main results of interest were CD4(+)-cell counts and odds ratios (ORs) of low CD4(+)-cell counts, defined as below 350 cells per microl. CD4(+)-cell counts were similar in men and nonpregnant women, with a mean (standard deviation) of 828 (203) cells per microl, but pregnant women had a lower value of 771 (250) cells per microl. None of the factors assessed was related to the odds of having a low CD4(+)-cell count among men and nonpregnant women, but age, age of marriage, and alcohol usage were significant predictors in pregnant women. Compared to pregnant women less than 20 years old, older women had significantly lower odds of a low CD4(+)-cell count (ORs were 0.06 for women aged 20 to 29 years and 0.22 for those aged 30 to 39 years). When compared with those pregnant women who were married before 20 years of age, those who married at 20 to 29 years and 30 to 39 years had odds ratios of 6.41 and 9.40, respectively. Previous alcohol use was also associated with low CD4(+)-cell counts (OR, 5.15). The 95% confidence interval for CD4(+)-cell counts in healthy adult Nigerians is 547 to 1,327 cells per microl, and this is the first time this has been determined. PMID- 15817762 TI - Differential detection of five mouse-infecting helicobacter species by multiplex PCR. AB - Several species of helicobacter have been isolated from laboratory mice, including H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, which appear to be the most common. The most widely used published method for molecular detection of these agents is PCR amplification of a conserved region of 16S rRNA, but differential speciation requires restriction enzyme digestion of the amplicons. This study was undertaken to determine PCR conditions that would simultaneously and specifically identify each of the five common species without restriction enzyme analyses. First, we designed novel and specific PCR primers for H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, using sequences from the heterologous regions of 16S rRNA. Because of comigration of amplified products, we next identified P17, an H. bilis-specific protein; P25, an H. hepaticus-specific protein; and P30, an H. muridarum-specific protein by screening genomic DNA expression libraries of each species. Primers were designed from these three genes, plus newly designed, species-specific 16S rRNA primers for H. rodentium and H. typhlonius that could be utilized for a five-plex PCR. The sizes of the amplicons from H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius were 435, 705, 807, 191, and 122 bp, respectively, allowing simultaneous detection and effective discrimination among species. PMID- 15817763 TI - Real-time reverse transcription-PCR quantitation of substance P receptor (NK-1R) mRNA. AB - The substance P (SP)-preferring receptor, neurokinin-1 receptor (NK-1R), has an important role in inflammation, immune regulation, and viral infection. We applied a newly developed real-time reverse transcription (RT)-PCR assay to quantify NK-1R mRNA in human neuronal cell line (NT-2N), a human B-cell line (IM9), monocyte-derived macrophages (MDM), peripheral blood lymphocytes (PBL), and human astroglioma cells (U87 MG). The NK-1R real-time RT-PCR assay has a sensitivity of 100 mRNA copies, with a dynamic range of detection between 10(2) and 10(7) copies of NK-1R gene transcripts per reaction. This assay is highly reproducible, with an intraassay coefficient variation of threshold cycle (Ct) of less than 1.9%. The NK-1R real-time RT-PCR is highly sensitive for quantitative determination of NK-1R mRNA in human immune cells (MDM and PBL) that express low levels of NK-1R mRNA. In addition, the assay has the ability to accurately quantitate the dynamic changes in NK-1R mRNA expression in interleukin-1beta stimulated U87 MG. These data indicate that the NK-1R real-time RT-PCR has potential for a wide application in investigation of NK-1R expression at the mRNA level under physiological and pathological conditions in both the central nervous system and the immune system. PMID- 15817764 TI - Rapid competitive enzyme-linked immunosorbent assay for detection of antibodies to peste des petits ruminants virus. AB - Peste des petits ruminants (PPR) is a contagious viral disease of small ruminants that is of economic importance in Africa, the Middle East, and Asia. We developed a rapid competitive enzyme-linked immunosorbent assay (rapid c-ELISA) for the diagnosis and surveillance of PPR. This assay detects PPR virus (PPRV) antibodies in serum samples by quantifying the amount of monoclonal antibody (MAb) P-3H12 after 30 min of incubation of a serum-MAb conjugate mixture on plates coated with a PPRV recombinant nucleocapsid protein (rPPRV-N). We tested 249 PPRV-positive serum samples and 733 PPRV-negative serum samples from field ruminants. The threshold of percent inhibition (PI) was determined to be <50 on the basis of the mean PI plus 3 standard deviations for sera from PPRV-negative ruminants. The relative specificity and sensitivity of the rapid c-ELISA were 98.5% (722 of 733 serum samples) and 93.4% (234 of 249 serum samples), respectively. The rapid c ELISA sensitively detected PPRV antibodies in hyperimmune sera (virus neutralization test [VNT] titer, >512), even at dilutions > or = 512 in normal goat serum, and as early as 6 to 13 days postinfection from 12 goats, each of which was infected with one of the four PPRV lineages. Hyperimmune sera from animals experimentally vaccinated with rinderpest virus gave positive results by the rapid c-ELISA when the rinderpest virus VNT titers were >512, although the rapid c-ELISA titers were very low (2 to 16). However, the rapid c-ELISA was negative when the rinderpest virus VNT titer was < or = 128. The rapid c-ELISA developed in the present work provides a short turnaround time and could be a useful tool for the diagnosis of PPR and screening for PPRV in the field. PMID- 15817765 TI - Development and evaluation of a Western blot kit for diagnosis of schistosomiasis. AB - We evaluated the performance of Western blot (WB) analysis using commercially available antigen strips and compared the results with those of indirect hemagglutination (IHA) and indirect immunofluorescence (IFAT) for the serodiagnosis of human schistosomiasis. The antigen preparation was a crude extract of Schistosoma mansoni. The WB profile characteristics of schistosomiasis were characterized by comparing the results for 58 serum samples from patients with parasitologically proven S. mansoni (n = 12) and S. haematobium (n = 46) infections and 37 individuals with probable cases of schistosomiasis but with only positive serology results. The specificity of WB analysis was assessed by testing 12 serum samples from healthy subjects, 67 serum samples from patients with other proven helminthic and protozoan infections, and 16 serum samples from patients with autoantibodies. Six immunodominant bands (65, 70, 80, 95, 110, and 120 kDa) were revealed with sera from patients with schistosomiasis. The presence of three or more bands in the range 65 to 120 kDa, with the exception of the 100 kDa band, was considered diagnostic for Schistosoma infection and had a specificity of 100% in our series. In patients with proven schistosomiasis, the sensitivity of WB analysis was 84.5%, whereas those of IFAT and IHA were 65.5 and 72.9%, respectively. For serologically proven cases, the sensitivity of WB analysis was 97.3%. The overall sensitivity and specificity for both groups of patients were 89.5 and 100%, respectively, with positive and negative predictive values of 100 and 91.3%, respectively. We conclude that WB analysis is a useful technique for the immunological diagnosis of schistosomiasis. PMID- 15817766 TI - Retrospective serological investigation of severe acute respiratory syndrome coronavirus antibodies in recruits from mainland China. AB - Different assays were used to analyze 1,621 serum specimens collected from military recruits from the People's Republic of China in 2002 for severe acute respiratory syndrome (SARS) coronavirus antibodies. The results demonstrated that the subjects either had rarely been exposed to the virus before the 2003 SARS outbreak or had not been exposed but the nucleocapsid protein cross-reacted with other antibodies in humans. PMID- 15817767 TI - Rapid immunofluorescence microscopy for diagnosis of melioidosis. AB - An immunofluorescent (IF) method that detects Burkholderia pseudomallei in clinical specimens within 10 min was devised. The results of this rapid method and those of an existing IF method were prospectively compared with the culture results for 776 specimens from patients with suspected melioidosis. The sensitivities of both IF tests were 66%, and the specificities were 99.5 and 99.4%, respectively. PMID- 15817768 TI - Immunization with recombinant surface antigen P50 of Babesia gibsoni expressed in insect cells induced parasite growth inhibition in dogs. AB - This is a report of a vaccine trial directed against Babesia gibsoni infection in dogs with the use of the recombinant antigen P50. Dogs immunized with P50 showed partial protection manifested as a significantly low level of parasitemia. The results indicated that P50 is a primary vaccine candidate molecule against canine B. gibsoni infection. PMID- 15817769 TI - Oxcarbazepine-induced immunoglobulin deficiency. PMID- 15817770 TI - Genome Update: Protein secretion systems in 225 bacterial genomes. PMID- 15817771 TI - Cataloguing our current genome collection. PMID- 15817772 TI - The epipolythiodioxopiperazine (ETP) class of fungal toxins: distribution, mode of action, functions and biosynthesis. AB - Epipolythiodioxopiperazines (ETPs) are toxic secondary metabolites made only by fungi. The best-known ETP is gliotoxin, which appears to be a virulence factor associated with invasive aspergillosis of immunocompromised patients. The toxicity of ETPs is due to the presence of a disulphide bridge, which can inactivate proteins via reaction with thiol groups, and to the generation of reactive oxygen species by redox cycling. With the availability of complete fungal genome sequences and efficient gene-disruption techniques for fungi, approaches are now feasible to delineate biosynthetic pathways for ETPs and to gain insights into the evolution of such gene clusters. PMID- 15817773 TI - The Pbs2 MAP kinase kinase is essential for the oxidative-stress response in the fungal pathogen Candida albicans. AB - The human fungal pathogen Candida albicans responds to stress by phosphorylation of the Hog1 MAP kinase. PBS2 was cloned and shown to encode the MAP kinase kinase that is involved in this activation, as determined by immunoblot analyses using antibodies that recognize the active form of the target Hog1 protein. Characterization of pbs2 mutants revealed that they were sensitive to both osmotic and oxidative stress and that they, interestingly, displayed differential behaviour from that of hog1 mutants, losing viability when exposed to an oxidative challenge more rapidly than the hog1 strain. Hog1 and Pbs2 were also shown to be involved in the mechanism of adaptation to oxidative stress, as evidenced by the enhanced susceptibility to oxidants of pbs2 and hog1 mutants, compared with the wild-type strain, when cells were previously exposed to a low, sub-lethal concentration of hydrogen peroxide and by the PBS2-dependent diminished activation of Hog1 MAP kinase in the adaptive process. Studies with a chimaeric Hog1-green fluorescent protein fusion revealed that this protein was localized throughout the cell (being excluded from the vacuole), but concentrated in the nucleus in response to NaCl stress, a process that was dependent on the Pbs2 protein. Both Hog1 and Pbs2 also play a role in controlling the phosphorylation state of the other MAP kinases Mkc1 and Cek1, involved respectively in cell-wall integrity and invasive growth. Furthermore, it is demonstrated that PBS2 plays a role in cell-wall biogenesis in this fungal pathogen, as its deletion renders cells with an altered susceptibility to certain cell wall-interfering compounds. PMID- 15817774 TI - Construction and real-time RT-PCR validation of Candida albicans PALS-GFP reporter strains and their use in flow cytometry analysis of ALS gene expression in budding and filamenting cells. AB - The gene encoding yeast-enhanced green fluorescent protein (GFP) was placed under control of ALS gene promoters in Candida albicans. The PALS-GFP reporter strains were validated using various techniques including a new real-time RT-PCR assay to quantify ALS gene expression. The PALS-GFP reporter strains were grown in media that promoted yeast or germ tube forms, and the resulting fluorescence was measured by flow cytometry. In addition to results that indicate differences in ALS gene expression due to growth medium, growth stage and developmental programme, new data show large differences in transcriptional level among the ALS genes. Expression of ALS1 was associated with transfer of the PALS1-GFP strain to fresh growth medium. ALS3 expression increased markedly when germ tubes were visible microscopically and ALS7 expression exhibited a transient peak between 2 and 3 h following inoculation into fresh YPD medium. Transcription from the ALS1 and ALS3 promoters was strongest among those tested and contrasted markedly with the weaker promoter strength at the ALS5, ALS6, ALS7 and ALS9 loci. These weaker transcriptional responses were also observed using real-time RT-PCR measurements on wild-type C. albicans cells. Assuming a positive correlation between transcriptional level and protein production, these results suggest that some Als proteins are abundant on the C. albicans cell surface while others are produced at a much lower level. PMID- 15817775 TI - Flanking direct repeats of hisG alter URA3 marker expression at the HWP1 locus of Candida albicans. AB - HWP1 encodes an adhesin of Candida albicans and has been implicated in filamentation and virulence. URA3, an often-used transformation selection marker, is apparently incorrectly expressed when integrated at the HWP1 locus, which results in an attenuated virulence phenotype. Expression of URA3 is compromised by ectopic integration at other loci as well. In contrast, prior studies from the authors' laboratory had demonstrated that the filamentation deficiency and attenuated virulence of hwp1Delta mutants were fully restored in rescued strains in which URA3 was integrated at the HWP1 locus. This discrepancy prompted a reinvestigation of these mutants. A series of congenic strains were constructed which demonstrated that the filamentation and virulence defects of a homozygous hwp1Delta mutant could be rescued without introduction of a functional HWP1 allele. Despite the absence of detectable differences in URA3 expression, analysis of suppressor mutations suggested that reduced URA3 expression gave rise to the mutant phenotypes. Several independent spontaneous suppressor mutations that restored filamentation to strains of genotype hwp1Delta : : hisG-URA3 hisG/hwp1Delta : : hisG had acquired a tandem duplication of the hisG-URA3-hisG marker cassette. The hwp1 null mutant and rescued strains differed by the presence or absence of flanking hisG sequence. Substitution of the hisG-URA3-hisG insert of the hwp1 null mutant with URA3 alone largely rescued the filamentation and virulence phenotypes. The presence of a single copy of hisG adjacent to URA3 had no effect. It is concluded that flanking direct repeats of hisG, present as part of a recyclable disruption cassette, negatively influenced URA3 expression and are responsible for the previously reported phenotypes of the hwp1 mutants. PMID- 15817776 TI - Phenotype switching affects biofilm formation by Candida parapsilosis. AB - Generation of biofilms by the pathogenic yeast Candida parapsilosis is correlated closely with disease. The phenomenon of phenotype switching in 20 isolates of C. parapsilosis was examined and the relationship with biofilm development was investigated. Four stable and heritable phenotypes were identified--crepe, concentric, smooth and crater. Cells from crepe and concentric phenotypes are almost entirely pseudohyphal, whilst cells from smooth and crater phenotypes are mostly yeast-like. The pseudohyphae from concentric phenotypes are approximately 45 % wider than those from crepe cells. The cell size of the smooth phenotype is smaller than those of the other three phenotypes. On polystyrene surfaces, the concentric phenotype generates up to twofold more biofilm than the crepe and crater phenotypes. Smooth phenotypes generate the least biofilm. Concentric phenotypes also invade agar surfaces more than the crepe and crater phenotypes, whilst smooth phenotypes do not invade at all. The smooth phenotype, however, grows significantly faster than the others. The quorum-sensing molecule farnesol inhibits formation of biofilms by the crepe, concentric and crater phenotypes. PMID- 15817777 TI - Elucidation of stability determinants of cold-adapted monomeric isocitrate dehydrogenase from a psychrophilic bacterium, Colwellia maris, by construction of chimeric enzymes. AB - To elucidate determinants of differences in thermostability between mesophilic and psychrophilic monomeric isocitrate dehydrogenases (IDHs) from Azotobacter vinelandii (AvIDH) and Colwellia maris (CmIDH), respectively, chimeric enzymes derived from the two IDHs were constructed based on the recently resolved three dimensional structure of AvIDH, and several characteristics of the two wild-type and six chimeric IDHs were examined. These characteristics were then compared with those of dimeric IDH from Escherichia coli (EcIDH). All recombinant enzymes with a (His)(6)-tag attached to the N-terminal were overexpressed in the E. coli cells and purified by Ni(2+)-affinity chromatography. The catalytic activity (k(cat)) and catalytic efficiency (k(cat)/K(m)) of the wild-type AvIDH and CmIDH were higher than those of EcIDH, implying that an improved catalytic rate more than compensates for the loss of a catalytic site in the former two IDHs due to monomerization. Analyses of the thermostability and kinetic parameters of the chimeric enzymes indicated that region 2, corresponding to domain II, and particularly region 3 located in the C-terminal part of domain I, are involved in the thermolability of CmIDH, and that the corresponding two regions of AvIDH are important for exhibiting higher catalytic activity and affinity for isocitrate than CmIDH. The relationships between the stability, catalytic activity and structural characteristics of AvIDH and CmIDH are discussed. PMID- 15817778 TI - The 120 592 bp IncF plasmid pRSB107 isolated from a sewage-treatment plant encodes nine different antibiotic-resistance determinants, two iron-acquisition systems and other putative virulence-associated functions. AB - The antibiotic-multiresistance IncF plasmid pRSB107 was isolated by a transformation-based approach from activated-sludge bacteria of a wastewater treatment plant. It confers resistance to ampicillin, penicillin G, chloramphenicol, erythromycin, kanamycin, neomycin, streptomycin, sulfonamides, tetracycline and trimethoprim and against mercuric ions. Complete sequencing of this plasmid revealed that it is 120 592 bp in size and has a G+C content of 53.1 mol%. The plasmid backbone is composed of three replicons, RepFIA, RepFIB and RepFII, which are almost identical to corresponding regions located on the F plasmid and on R100. The three replicons encode replication initiation (rep) and replication control, multimer resolution (mrs), post-segregational killing of plasmid-free cells (psk) and active plasmid partitioning (sopABC locus). Part of the F-leading region and remnants of the F-homologous DNA-transfer (tra) module complete the pRSB107 backbone. Plasmid pRSB107 contains a complex, highly mosaic 35 991 bp antibiotic-resistance region consisting of a Tn21- and a Tn10 derivative and a chloramphenicol-resistance module. The Tn21 derivative is composed of a mercury-resistance region (mer), a Tn4352B-like kanamycin/neomycin resistance transposon, a streptomycin/sulfonamide-resistance module, remnants of the beta-lactam-resistance transposon Tn1, a macrolide-resistance module flanked by copies of IS26 and IS6100, remnants of Tn402 integrating a class 1 integron and the Tn21-specific transposition module. A truncated version of the tetracycline-resistance transposon Tn10 and the chloramphenicol acetyltransferase gene catA complete the pRSB107 resistance region. In addition to antibiotic resistance, pRSB107 encodes the following putative virulence-associated functions: (i) an aerobactin iron-acquisition siderophore system (iuc/iut); (ii) a putative high-affinity Fe(2+) uptake system which was previously identified on a pathogenicity island of Yersinia pestis and in the genome of the phytopathogen Erwinia carotovora subsp. atroseptica SCRI1043; (iii) an sn-glycerol-3-phosphate transport system (ugp); and (iv) the virulence-associated genes vagCD having a possible function in stable plasmid inheritance. All the accessory modules are framed by insertion sequences, indicating that pRSB107 was gradually assembled by integration of different horizontally acquired DNA segments via transposition or homologous recombination. PMID- 15817779 TI - The MexGHI-OpmD multidrug efflux pump controls growth, antibiotic susceptibility and virulence in Pseudomonas aeruginosa via 4-quinolone-dependent cell-to-cell communication. AB - In Pseudomonas aeruginosa the production of multiple virulence factors depends on cell-to-cell communication through the integration of N-acylhomoserine lactone (AHL)- and 2-heptyl-3-hydroxy-4(1H)-quinolone (PQS)- dependent signalling. Mutation of genes encoding the efflux protein MexI and the porin OpmD from the MexGHI-OpmD pump resulted in the inability to produce N-(3-oxododecanoyl)-L homoserine lactone (3-oxo-c12-hsl) and pqs and a marked reduction in n-butanoyl-L homoserine lactone levels. Both pump mutants were impaired in growth and exhibited enhanced rather than reduced antibiotic resistance. Provision of exogenous PQS improved growth and restored AHL and virulence factor production as well as antibiotic susceptibility, indicating that the pump mutants retained their capacity to respond to PQS. RT-PCR analysis indicated that expression of the PQS biosynthetic genes, phnA and pqsA, was inhibited when the mutants reached stationary phase, suggesting that the pleiotropic phenotype observed may be due to intracellular accumulation of a toxic PQS precursor. To explore this hypothesis, double mexI phnA (unable to produce anthranilate, the precursor of PQS) and mexI pqsA mutants were constructed; the improved growth of the former suggested that the toxic compound is likely to be anthranilate or a metabolite of it. Mutations in mexI and opmD also resulted in the attenuation of virulence in rat and plant infection models. In plants, addition of PQS restored the virulence of mexI and opmD mutants. Collectively, these results demonstrate an essential function for the MexGHI-OpmD pump in facilitating cell-to-cell communication, antibiotic susceptibility and promoting virulence and growth in P. aeruginosa. PMID- 15817780 TI - Expression of the quorum-sensing regulatory protein LasR is strongly affected by iron and oxygen concentrations in cultures of Pseudomonas aeruginosa irrespective of cell density. AB - The expression of the transcriptional regulatory protein LasR, a main component of the quorum-sensing (QS) system in Pseudomonas aeruginosa, was recently found to be sensitive to several environmental factors in addition to its dependency on cell density. However, the inherent effects of the different factors have seldom been separately demonstrated due to concurrent changes of culture conditions in typical experimental settings. Furthermore, the interplays of the different factors are unknown. In this work, the effects and interplay of iron concentration and dissolved oxygen tension (pO(2)) on the expression of lasR in P. aeruginosa were studied in defined growth media with varied iron concentration and pO(2) values in computer-controlled batch and continuous cultures. beta Galactosidase activity in a recombinant P. aeruginosa PAO1 (NCCB 2452) strain with a lasRp-lacZ fusion was used as a reporter for lasR expression. In batch culture with a constant pO(2) approximately 10 % air saturation, a strong correlation between the exhaustion of iron and the increase of lasR expression was observed. In continuous culture with nearly constant cell density but varied pO(2) values, lasR expression generally increased with increasing oxidative stress with the exception of growth under O(2)-limited conditions (pO(2) approximately equal to 0 %). Under O(2) limitation, the expression of lasR strongly depended on the concentration of iron. It showed a nearly twofold increase in cells grown under iron deprivation in comparison with cells grown in iron-replete conditions and reached the expression level seen at high oxidative stress. A preliminary proteomic analysis was carried out for extracellular proteins in samples from batch cultures grown under different iron concentrations. Several of the extracellular proteins (e.g. AprA, LasB, PrpL) which were up-regulated under iron-limited conditions were found to be QS regulated proteins. Thus, this study clearly shows the links between QS and genes involved in iron and oxygen regulation in P. aeruginosa. PMID- 15817781 TI - Protein expression by a Beijing strain differs from that of another clinical isolate and Mycobacterium tuberculosis H37Rv. AB - The Beijing strain family has often been associated with tuberculosis (TB) outbreaks and drug resistance worldwide. In this study the authors have compared the protein expression and antigen recognition profiles of a local Beijing strain with a less prevalent clinical isolate belonging to the family 23 strain lineage, and the laboratory strain Mycobacterium tuberculosis H37Rv. Using two-dimensional electrophoresis, liquid chromatography tandem mass spectrometry and Western blot analysis several proteins were identified as quantitatively increased or decreased in both clinical strains compared to H37Rv. Remarkably, the Beijing strain showed increased expression of alpha-crystallin and decreased expression of Hsp65, PstS1, and the 47 kDa protein compared to the other clinical strain and H37Rv. One- and two-dimensional Western blot analysis of antigens expressed by the three strains, using plasma from TB patients, confirmed differential antigen expression by strains and patient-to-patient variation in humoral immunity. These observed protein differences could aid the elucidation of mechanisms underlying the success of the Beijing strain family, measured by global dissemination, compared to other M. tuberculosis strains. PMID- 15817782 TI - Identification of a conserved Moraxella catarrhalis haemoglobin-utilization protein, MhuA. AB - Moraxella catarrhalis is a leading cause of acute otitis media in children and is a cause of respiratory disease in adults with underlying lung disease. This organism is a strict human pathogen that has an absolute requirement for iron in order to grow and cause disease. Previous studies identified transferrin and lactoferrin receptors used by M. catarrhalis to obtain iron from the human host, yet other iron-acquisition systems remain undefined. In this study, it is demonstrated that this strict mucosal pathogen can utilize haemoglobin (Hb) as a sole source of iron for growth. A novel 107 kDa outer-membrane protein involved in Hb utilization by this pathogen was also identified. An isogenic mutant defective in this Moraxella Hb-utilization protein (MhuA), 7169 : : mhuA, showed a significant lag during growth in the presence of Hb as the sole iron source. This protein appears to be expressed constitutively, regardless of growth conditions, and a mAb directed to MhuA demonstrated that this protein contains highly conserved, surface-exposed epitopes. Data demonstrating that expression of MhuA may be highly specific to isolates of M. catarrhalis are also presented, suggesting a potential role as a diagnostic marker. To our knowledge, this is the first report demonstrating that M. catarrhalis expresses an Hb-binding protein and that this bacterium can utilize Hb as a sole iron source for growth. PMID- 15817783 TI - PrpZ, a Salmonella enterica serovar Typhi serine/threonine protein phosphatase 2C with dual substrate specificity. AB - Genes encoding eukaryotic-type protein kinases and phosphatases are present in many bacterial genomes. An ORF encoding a polypeptide with homology to protein phosphatases 2C (PP2Cs) was identified in the genomes of Salmonella enterica serovar Typhi strains CT18 and Ty2. This protein, termed PrpZ, is the first PP2C to be identified in enterobacteria. Analysis of the amino acid sequence revealed two distinct domains: the N-terminal segment containing motifs of the catalytic domain of PP2Cs and the C-terminal segment with unknown function. PrpZ was expressed in Escherichia coli as a histidine-tagged fusion protein (PrpZ(His)) and the purified protein was analysed for its ability to dephosphorylate various substrates. Using p-nitrophenyl phosphate as a substrate, optimal PrpZ(His) activity was observed at pH 9.5, with a strong preference for Mn(2+) over Mg(2+). Activity of PrpZ(His) was inhibited by EDTA, sodium fluoride, sodium phosphate and sodium pyrophosphate but unaffected by okadaic acid, indicating that PrpZ is a PP2C. Using synthetic phosphopeptides as substrates, PrpZ(His) could hydrolyse phosphorylated serine, threonine or tyrosine residues, with the highest catalytic efficiency (k(cat)/K(m)) for the threonine phosphopeptide. With phosphorylated myelin basic protein (MBP) as the substrate, Mn(2+) was only twofold more efficient than Mg(2+) in stimulating PrpZ(His) activity at pH 8.0. The ability of PrpZ(His) to remove the phosphoryl group from phosphotyrosine residues was confirmed by measuring the release of inorganic phosphate from phospho-Tyr MBP. Together, these data indicate that PrpZ has all the features of a PP2C with dual substrate specificity in vitro. PMID- 15817784 TI - Biochemical evidence that the pduS gene encodes a bifunctional cobalamin reductase. AB - Salmonella enterica degrades 1,2-propanediol (1,2-PD) by a pathway that requires coenzyme B(12) (adenosylcobalamin; AdoCbl). The genes specifically involved in 1,2-PD utilization (pdu) are found in a large contiguous cluster, the pdu locus. Earlier studies have indicated that this locus includes genes for the conversion of vitamin B(12) (cyanocobalamin; CNCbl) to AdoCbl and that the pduO gene encodes an ATP : cob(I)alamin adenosyltransferase which catalyses the terminal step of this process. Here, in vitro evidence is presented that the pduS gene encodes a bifunctional cobalamin reductase that catalyses two reductive steps needed for the conversion of CNCbl into AdoCbl. The PduS enzyme was produced in high levels in Escherichia coli. Enzyme assays showed that cell extracts from the PduS expression strain reduced cob(III)alamin (hydroxycobalamin) to cob(II)alamin at a rate of 91 nmol min(-1) mg(-1) and cob(II)alamin to cob(I)alamin at a rate of 7.8 nmol min(-1) mg(-1). In contrast, control extracts had only 9.9 nmol min(-1) mg( 1) cob(III)alamin reductase activity and no detectable cob(II)alamin reductase activity. Thus, these results indicated that the PduS enzyme is a bifunctional cobalamin reductase. Enzyme assays also showed that the PduS enzyme reduced cob(II)alamin to cob(I)alamin for conversion into AdoCbl by purified PduO adenosyltransferase. Moreover, studies in which iodoacetate was used as a chemical trap for cob(I)alamin indicated that the PduS and PduO enzymes physically interact and that cob(I)alamin is sequestered during the conversion of cob(II)alamin to AdoCbl by these two enzymes. This is likely to be important physiologically, since cob(I)alamin is extremely reactive and would need to be protected from unproductive by-reactions. Lastly, bioinformatic analyses showed that the PduS enzyme is unrelated in amino acid sequence to enzymes of known function currently present in GenBank. Hence, results indicate that the PduS enzyme represents a new class of cobalamin reductase. PMID- 15817785 TI - ibeA, a virulence factor of avian pathogenic Escherichia coli. AB - The presence of ibeA, a gene encoding a known virulence factor of Escherichia coli strains responsible for neonatal meningitis in humans, was investigated in the genome of 213 avian pathogenic E. coli (APEC) strains and 55 non-pathogenic E. coli strains of avian origin. Fifty-three strains were found to be ibeA(+), all of which belonged to the APEC group. The ibeA gene is therefore positively linked to the pathogenicity of strains (P<0.0001). Analysis of the serogroup of strains revealed a positive association of ibeA with serogroups O18, O88 and O2. On the contrary, only 1/59 O78 strains are ibeA(+), indicating a negative association of ibeA with this serogroup (P<0.0001). The role of ibeA in the virulence of the APEC strain BEN 2908 was investigated by constructing an ibeA mutant. Challenge assays on 3-week-old chickens showed a reduced virulence for the ibeA mutant. Furthermore, the APEC strain BEN 2908 was able to invade brain microvascular epithelial cells, this invasion being significantly reduced upon inactivation of ibeA. Altogether, these results suggest a role of ibeA in the pathogenicity of some APEC strains and confirm the close relationship between APEC and other human extraintestinal pathogenic E. coli isolates. PMID- 15817786 TI - The expression profile of Escherichia coli K-12 in response to minimal, optimal and excess copper concentrations. AB - The gene expression profile of Escherichia coli K-12 MG1655 grown in minimal medium supplemented with elevated copper concentrations (as copper-glycine) has been analysed using whole-genome oligonucleotide microarrays. At 750 muM copper glycine, the expression of both the cue and cus copper-export systems is evident. At near-lethal copper concentrations (2 mM copper-glycine), the expression of these two regulons increases significantly. Other regulons with increased transcription in response to elevated concentrations of copper-glycine include those for the superoxide stress response, iron homeostasis, and envelope stress. Furthermore, a variety of ORFs with decreased expression in response to increased copper-glycine has been identified, including the zinc ABC transporter and genes involved in the chemotactic response. PMID- 15817787 TI - The H-NS protein represses transcription of the eltAB operon, which encodes heat labile enterotoxin in enterotoxigenic Escherichia coli, by binding to regions downstream of the promoter. AB - Heat-labile enterotoxin, a major virulence determinant of enterotoxigenic Escherichia coli, is encoded by the eltAB operon. To elucidate the molecular mechanism by which the heat-stable nucleoid-structural (H-NS) protein controls transcription of eltAB, the authors constructed an eltAB-lacZ transcriptional fusion and performed beta-galactosidase analysis. The results showed that H-NS protein exerts fivefold repression on transcription from the eltAB promoter at 37 degrees C and 10-fold repression at 22 degrees C. Two silencer regions that were required for H-NS-mediated repression of eltAB expression were identified, both of which were located downstream of the start site of transcription. One silencer was located between +31 and +110, the other between +460 and +556, relative to the start site of transcription, and they worked cooperatively in repression. DNA sequences containing the silencers were predicted to be curved by in silico analysis and bound H-NS protein directly in vitro. Repression of eltAB transcription by H-NS was independent of promoter strength, and the presence of H NS protein did not affect promoter opening in vitro, indicating that repression was achieved by inhibiting promoter clearance or blocking transcription elongation, probably via DNA looping between the two silencers. PMID- 15817788 TI - Generation of transposon insertion mutant libraries for Gram-positive bacteria by electroporation of phage Mu DNA transposition complexes. AB - Transposon mutagenesis is a powerful technique for generating collections of insertion mutants for genetic studies. This paper describes how phage Mu DNA transposition complexes, transpososomes, can be exploited for gene delivery to efficiently introduce selectable markers to genomes of Gram-positive bacteria. Mu transpososomes were assembled in vitro with custom-designed mini-Mu transposons, concentrated, and electroporated into cells of three Gram-positive bacterial species: Staphylococcus aureus, Streptococcus pyogenes and Streptococcus suis. Within cells, the complexes reproduced an authentic DNA transposition reaction and integrated the delivered transposons into the bacterial genomes, yielding single-copy insertions. The integration efficiency among different species and strains of Gram-positive bacteria ranged from 1x10(1) to 2x10(4) c.f.u. (mug introduced transposon DNA)(-1). The strategy should be applicable to a variety of other Gram-positive species after initial optimization of certain key factors affecting transposon delivery, such as the preparation method of competent cells and physical parameters of electroporation. This study extends the scope of the Mu transpososome delivery-based genomic DNA integration strategy to Gram-positive bacteria. Thus, a straightforward generation of sizeable mutant banks is feasible for these bacteria, potentiating several types of genomic-level approaches for studies of a variety of important bacterial processes, such as pathogenicity. PMID- 15817789 TI - Sequencing, characterization and transcriptional analysis of the histidine decarboxylase operon of Lactobacillus buchneri. AB - The amplification of an internal fragment of the hdcA gene for histidine decarboxylase in Lactobacillus buchneri showed the gene to be located on the bacterial chromosome. Reverse PCR was then used to amplify both it and its adjacent genes. The histidine decarboxylase cluster was found to be composed of four genes: hdcC (expressed in Lactococcus lactis, the product of which is located in the membrane, suggesting it to be a histidine/histamine antiporter), hdcA (which encodes histidine decarboxylase), hdcB (of unknown function but co transcribed as bicistronic mRNA together with hdcA) and hisS (the only copy of a gene encoding a histidyl-tRNA synthetase in Lb. buchneri). The expression of hisS depends on the histidine concentration of the growth medium, and it can be transcribed as monocistronic or hdcA-hdcB-hisS polycistronic mRNA. PMID- 15817790 TI - Genome-based in silico detection of putative manganese transport systems in Lactobacillus plantarum and their genetic analysis. AB - Manganese serves an important function in Lactobacillus plantarum in protection against oxidative stress and this bacterium can accumulate Mn(2+) up to millimolar levels intracellularly. Although the physiological role of Mn(2+) and the uptake of this metal ion have been well documented, the only uptake system described so far for this bacterium is the Mn(2+)- and Cd(2+)-specific P-type ATPase (MntA). Recently, the genome of L. plantarum WCFS1 has been sequenced allowing in silico detection of genes potentially encoding Mn(2+) transport systems, using established microbial Mn(2+) transporters as the query sequence. This genome analysis revealed that L. plantarum WCFS1 encodes, besides the previously described mntA gene, an ABC transport system (mtsCBA) and three genes encoding Nramp transporters (mntH1, mntH2 and mntH3). The expression of three (mtsCBA, mntH1 and mntH2) of the five transport systems was specifically derepressed or induced upon Mn(2+) limitation, supporting their role in Mn(2+) homeostasis in L. plantarum. However, in contrast to previous reports, mntA expression remains below detection levels in both Northern and real-time RT-PCR analysis in both Mn(2+) excess and starvation conditions. Growth of WCFS1 derivatives mutated in mntA, mtsA or mntH2, or both mtsA and mntH2 appears unaffected under Mn(2+) excess or Mn(2+) limitation. Moreover, intracellular Mn(2+) concentrations remained unaltered in these mutants compared to the wild type. This may suggest that this species is highly adaptive in response to inactivation of these genes or, alternatively, that other transporters that have not yet been identified as Mn(2+) transporters in bacteria are involved in Mn(2+) homeostasis in L. plantarum. PMID- 15817791 TI - The hydrogenases of Geobacter sulfurreducens: a comparative genomic perspective. AB - The hydrogenase content of the genome of Geobacter sulfurreducens, a member of the family Geobacteraceae within the delta-subdivision of the Proteobacteria, was examined and found to be distinct from that of Desulfovibrio species, another family of delta-Proteobacteria on which extensive research concerning hydrogen metabolism has been conducted. Four [NiFe]-hydrogenases are encoded in the G. sulfurreducens genome: two periplasmically oriented, membrane-bound hydrogenases, Hya and Hyb, and two cytoplasmic hydrogenases, Mvh and Hox. None of these [NiFe] hydrogenases has a counterpart in Desulfovibrio species. Furthermore, the large and small subunits of Mvh and Hox appear to be related to archaeal and cyanobacterial hydrogenases, respectively. Clusters encoding [Fe]-hydrogenases and periplasmic [NiFeSe]-hydrogenases, which are commonly found in the genomes of Desulfovibrio species, are not present in the genome of G. sulfurreducens. Hydrogen-evolving Ech hydrogenases, which are present in the genomes of at least two Desulfovibrio species, were also absent from the G. sulfurreducens genome, despite the fact that G. sulfurreducens is capable of hydrogen production. Instead, the G. sulfurreducens genome contained a cluster encoding a multimeric Ech hydrogenase related (Ehr) complex that was similar in content to operons encoding Ech hydrogenases, but did not appear to encode a hydrogenase. Phylogenetic analysis revealed that the G. sulfurreducens ehr cluster is part of a family of related clusters found in both the Archaea and Bacteria. PMID- 15817793 TI - Neurotoxins in axenic oscillatorian cyanobacteria: coexistence of anatoxin-a and homoanatoxin-a determined by ligand-binding assay and GC/MS. AB - Two neurotoxic alkaloids, anatoxin-a and its homologue homoanatoxin-a, were purified from the filamentous cyanobacteria Oscillatoria sp. strain 193 (PCC 9240) and Oscillatoria formosa NIVA CYA-92 (PCC 10111), respectively, and characterized by mass spectrometry. Biological activity was determined by examining the capacity of the toxins to competitively inhibit the binding of radiolabelled bungarotoxin to acetylcholine receptors, using post-synaptic membrane fractions of Torpedo electric tissue. Inhibition was concentration dependent, with a K(i) of 5.4+/-1.1x10(-8) M for anatoxin-a and 7.4+/-0.9x10(-8) M for homoanatoxin-a. Their high affinities for the nicotinic cholinergic receptors were exploited to adapt the radioligand-binding assay for routine detection of this class of neurotoxins directly in low-molecular-mass cell extracts of cyanobacteria. Confirmation of the results and toxin identification were achieved by coupled gas chromatography-mass spectrometry (GC/MS). Seventy six axenic strains, representative of 13 genera, were analysed. Five strains of the genus Oscillatoria, hitherto unknown for their toxicity, inhibited bungarotoxin binding. GC/MS revealed that Oscillatoria sp. strains PCC 6407, PCC 6412 and PCC 9107 synthesized exclusively anatoxin-a, whereas both anatoxin-a and homoanatoxin-a were produced by strain PCC 9029. Oscillatoria sp. strain PCC 6506, an isolate co-identic with strain PCC 9029, also produced both neurotoxins, but their respective presence depended upon growth conditions. The latter results suggest that regulatory differences in at least some of the cyanobacterial strains may account for the preferential synthesis of only one of the two neurotoxins or for their simultaneous occurrence. PMID- 15817792 TI - Biochemical and molecular characterization of a periplasmic hydrolase for oxidized polyvinyl alcohol from Sphingomonas sp. strain 113P3. AB - Oxidized polyvinyl alcohol hydrolase (OPH) and polyvinyl alcohol dehydrogenase were found to be constitutively present in the periplasm of Sphingomonas sp. strain 113P3 (formerly Pseudomonas sp. 113P3). The OPH was purified to homogeneity with a yield of 40 % and a 5.9-fold increase in specific activity. The enzyme was a homodimer consisting of 35 kDa subunits. Its activity was inhibited by PMSF, Hg(2+) and Zn(2+). The enzyme hydrolysed oxidized polyvinyl alcohol (oxidized PVA) and p-nitrophenyl acetate (PNPA), but did not hydrolyse any of the mono- or diketones tested. K(m) and V(max) values for oxidized PVA and PNPA were 0.2 and 0.3 mM, and 0.1 and 3.4 micromol min(-1) mg(-1), respectively. The gene for OPH was cloned and sequenced. Sequencing analysis revealed that the open reading frame consisted of 1095 bp, corresponding to a protein of 364 amino acids residues, encoding a signal peptide and a mature protein of 34 and 330 amino acids residues, respectively. The presence of a serine-hydrolase motif (a lipase box; Gly-X-Ser-X-Gly) strongly suggested that the enzyme belongs to the serine-hydrolase family. The protein exhibited homology with OPH of the Pseudomonas sp. strain VM15C (63 % identity) and the polyhydroxybutyrate depolymerases from Mesorhizobium loti, Rhizobium sp. and Sinorhizobium meliloti (29-32 % identity). The oph gene was expressed in Escherichia coli under the control of the lac promoter. The recombinant protein had the same molecular mass and N-terminal amino acid sequence as the purified OPH from strain 113P3. PMID- 15817794 TI - Protein phosphatase PphA from Synechocystis sp. PCC 6803: the physiological framework of PII-P dephosphorylation. AB - The phosphorylated signal transduction protein P(II) (P(II)-P) in the cyanobacterium Synechocystis sp. strain PCC 6803 is dephosphorylated by PphA, a protein phosphatase of the 2C family (PP2C). In this study, the physiological conditions of P(II)-P dephosphorylation were investigated with respect to the in vivo specificity of P(II)-P towards PphA and the cellular abundance of PphA in cells growing under different nitrogen regimes. Furthermore, the consequences of impaired P(II)-P dephosphorylation with respect to short-term inhibition of glutamine synthetase (GS) were studied. With a contribution of approximately 15 % of total Mn(2+)-dependent p-nitrophenyl phosphate hydrolysis activity, PphA has only a minor impact on the total PP2C activity in Synechocystis extracts. Nevertheless, residual P(II)-P dephosphorylation in PphA-deficient cells could only be observed after prolonged incubation in the presence of ammonium. The abundance of PphA correlates with the phosphorylation state of P(II) under nitrogen-replete conditions and is specifically enhanced by nitrite. Regulation of pphA expression operates at the post-transcriptional level. In the presence of nitrate/nitrite, PphA is present in molar excess over P(II)-P, enabling the cells to rapidly dephosphorylate P(II)-P in response to changing environmental conditions. A PphA-deficient mutant is not impaired in short-term inhibition of GS activity following ammonium treatment. Down-regulation of GS occurs by induction of gif genes (encoding GS inactivating factors 7 and 17), which is controlled by NtcA-mediated gene repression. Thus, impaired P(II)-P dephosphorylation does not affect ammonium-prompted inactivation of NtcA. PMID- 15817795 TI - Sulfoacetaldehyde is excreted quantitatively by Acinetobacter calcoaceticus SW1 during growth with taurine as sole source of nitrogen. AB - Eighteen enrichment cultures with taurine (2-aminoethanesulfonate) as the sole source of combined nitrogen under aerobic conditions were all successful, and 24 pure cultures were obtained. Only three of the cultures yielded an inorganic product, sulfate, from the sulfonate moiety of taurine, and the others were presumed to yield organosulfonates. Sulfoacetate, known from Rhodopseudomonas palustris CGA009 under these conditions, was not detected in any culture, but sulfoacetaldehyde (as a hydrazone derivative) was tentatively detected in the outgrown medium of nine isolates. The compound was stable under these conditions and the identification was confirmed by MALDI-TOF-MS. Most sulfoacetaldehyde releasing isolates were determined to be strains of Acinetobacter calcoaceticus, and a representative organism, strain SW1, was chosen for further work. A quantitative enzymic determination of sulfoacetaldehyde and its bisulfite addition complex was developed: it involved the NAD-coupled sulfoacetaldehyde dehydrogenase from R. palustris. A. calcoaceticus SW1 utilized taurine quantitatively and concomitantly with growth in, for example, an adipate-salts medium, and the release of sulfoacetaldehyde was stoichiometric. The deamination reaction involved a taurine dehydrogenase. Enrichment cultures to explore the possible release of organophosphonates from the analogous substrate, 2 aminoethanephosphonate, led to 33 isolates, all of which released inorganic phosphate quantitatively. PMID- 15817796 TI - The Ruminococcus albus pilA1-pilA2 locus: expression and putative role of two adjacent pil genes in pilus formation and bacterial adhesion to cellulose. AB - Ruminococcus albus produces fimbria-like structures that are involved with the bacterium's adhesion to cellulose. The subunit protein has been identified in strain 8 (CbpC) and strain 20 (GP25) and both are type IV fimbrial (Pil) proteins. The presence of a pil locus that is organized similarly in both strains is reported here together with the results of an initial examination of a second Pil protein. Downstream of the cbpC/gp25 gene (hereafter referred to as pilA1) is a second pilin gene (pilA2). Northern blot analysis of pilA1 and pilA2 transcripts showed that the pilA1 transcript is much more abundant in R. albus 8, and real-time PCR was used to measure pilA1 and pilA2 transcript abundance in R. albus 20 and its adhesion-defective mutant D5. Similar to the findings with R. albus 8, the relative expression of pilA1 in the wild-type strain was 73-fold higher than that of pilA2 following growth with cellobiose, and there were only slight differences between the wild-type and mutant strain in pilA1 and pilA2 transcript abundances, indicating that neither pilA1 nor pilA2 transcription is adversely affected in the mutant strain. Western immunoblots showed that the PilA2 protein is localized primarily to the membrane fraction, and the anti-PilA2 antiserum does not inhibit bacterial adhesion to cellulose. These results suggest that the PilA2 protein plays a role in the synthesis and assembly of type IV fimbriae-like structures by R. albus, but its role is restricted to cell associated functions, rather than as part of the externalized fimbrial structure. PMID- 15817797 TI - Sau42I, a BcgI-like restriction-modification system encoded by the Staphylococcus aureus quadruple-converting phage Phi42. AB - The serotype F phage Phi42 of Staphylococcus aureus is a triple-converting bacteriophage that encodes the staphylokinase gene (sak) and the enterotoxin A gene (entA). Lysogeny results in loss of expression of the chromosomal beta haemolysin gene (hlb) (negative conversion), the expression of staphylokinase and enterotoxin A (positive conversion), and the acquisition of resistance to lysis by all 23 phages of the International Basic Set (IBS) of S. aureus typing phages. Until this study, the basis of Phi42 resistance to lysis by exogenous phages was unknown. The authors report here that phage Phi42 encodes a restriction modification (R-M) system, termed Sau42I, adjacent to and in the same orientation to the phage integrase gene int. The genes encoding Sau42I were cloned and sequenced, and found to consist of two overlapping reading frames, ORF S (specificity) and ORF RM (restriction-modification), in the same orientation. The ORFs share a high degree of DNA and amino acid sequence homology with the previously characterized BcgI R-M system of Bacillus coagulans. Expression of the cloned Sau42I ORF S and ORF RM in S. aureus 80CR3 transformants from a plasmid vector conferred resistance to lysis by all 23 IBS phages. Similarly, transformants of S. aureus RN4220 harbouring recombinant plasmids containing both ORFs were resistant to lysis by the IBS typing phages. However, transformants harbouring plasmids encoding either ORF S or ORF RM were susceptible to lysis by the IBS phages, and they had the same phage-susceptibility pattern as the respective parental isolates. In vitro analysis of crude and partially purified extracts of S. aureus transformants harbouring both the Phi42 ORF S and ORF RM genes indicated that Sau42I has endonuclease activity and requires co-factors Mg(2+) and S-adenosylmethionine in order to function, and activity is optimized at pH 8, although the precise recognition sequence has yet to be determined. The findings of this study confirm that Phi42 is a quadruple-converting phage, believed to be the first described for S. aureus, and show that it encodes a novel R-M system termed Sau42I. PMID- 15817798 TI - Antiproliferative effects of phosphodiesterase type 5 inhibition in human pulmonary artery cells. AB - RATIONALE: Phosphodiesterase Type 5 (PDE5) inhibition represents a novel strategy for the treatment of pulmonary hypertension. OBJECTIVES: Our aim was to establish the distribution of PDE5 in the pulmonary vasculature and effects of PDE5 inhibition on pulmonary artery smooth muscle cells (PASMCs). METHODS AND MEASUREMENTS: PDE5 expression was examined by immunohistochemistry and Western blotting, in both normal and hypertensive lung tissues. DNA synthesis, proliferation, PDE activity, and apoptosis were measured in distal human PASMCs treated with soluble guanylyl cyclase activators (nitric oxide donors and BAY41 2272) and sildenafil. MAIN RESULTS: Cells containing PDE5 and alpha-smooth muscle actin occurred throughout the pulmonary vasculature, including obstructive intimal lesions. Three molecular forms of PDE5 were identified and protein expression was greater in hypertensive than control lung tissue. Most cyclic guanosine monophosphate hydrolysis (about 80%) in cultured cells was attributed to PDE5. Sildenafil induced a greater elevation of intracellular cyclic guanosine monophosphate levels compared with nitric oxide donors and BAY41-2272 (about 10 fold versus about 2-fold) and cotreatment had a synergistic effect, increasing cyclic nucleotide levels up to 50-fold. Dual stimulation of soluble guanylyl cyclase and inhibition of PDE5 activities also had significant downstream effects, increasing phosphorylation of vasodilator-stimulated phosphoprotein, reducing DNA synthesis and cell proliferation, and stimulating apoptosis, and these effects were mimicked by cyclic guanosine monophosphate analogs. CONCLUSIONS: Phosphodiesterase Type 5 is the main factor regulating cyclic guanosine monophosphate hydrolysis and downstream signaling in human PASMCs. The antiproliferative effects of this signaling pathway may be significant in the chronic treatment of pulmonary hypertension with PDE5 inhibitors such as sildenafil. PMID- 15817799 TI - Variation in ITGB3 is associated with asthma and sensitization to mold allergen in four populations. AB - RATIONALE: Recent genetic studies have implicated integrins in asthma and atopy susceptibility. We therefore evaluated the integrin-beta3 gene (ITGB3), an integrin gene within an asthma linkage peak on chromosome 17, as a candidate for susceptibility to asthma- and atopy-related phenotypes. METHODS AND MEASUREMENTS: We genotyped and performed association tests on 19 single nucleotide polymorphisms in ITGB3 in the Hutterites, a founder population, and in three outbred replication populations. MAIN RESULTS: Variation in ITGB3 was strongly associated with susceptibility to bronchial hyperresponsiveness and protection from allergic sensitization to mold allergens in this population. Three independent case-control populations representing Caucasians and African Americans were used to replicate this finding, also revealing ITGB3 alleles that are associated with asthma susceptibility and protection from mold allergen sensitization. CONCLUSIONS: This study provides evidence that ITGB3 plays a role in the pathogenesis of asthma and sensitization to mold allergens. PMID- 15817801 TI - A century of control of breathing. PMID- 15817800 TI - The mysterious pulmonary brush cell: a cell in search of a function. AB - Brush cells, also termed tuft, caveolated, multivesicular, and fibrillovesicular cells, are part of the epithelial layer in the gastrointestinal and respiratory tracts. The cells are characterized by the presence of a tuft of blunt, squat microvilli (approximately 120-140/cell) on the cell surface. The microvilli contain filaments that stretch into the underlying cytoplasm. They have a distinctive pear shape with a wide base and a narrow microvillous apex. The function of the pulmonary brush cell is obscure. For this reason, a working group convened on August 23, 2004, in Bethesda, Maryland, to review the physiologic role of the brush (microvillous) cell in normal airways and alveoli and in respiratory diseases involving the alveolar region (e.g., emphysema and fibrosis) and airway disease characterized by either excessive or insufficient amounts of airway fluid (e.g., cystic fibrosis, chronic bronchitis, and exercise-induced asthma). The group formulated several suggestions for future investigation. For example, it would be useful to have a panel of specific markers for the brush cell and in this way separate these cells for culture and more direct examination of their function (e.g., microarray analysis and proteomics). Using quantitative analysis, it was suggested to examine the number and location of the cells in disease models. Understanding the function of these cells in alveoli and airways may provide clues to the pathogenesis of several disease states (e.g., cystic fibrosis and fibrosis) as well as a key for new therapeutic modalities. PMID- 15817802 TI - Oxygenation index predicts outcome in children with acute hypoxemic respiratory failure. AB - To define outcome and time dependence of predictors of outcome in pediatric acute hypoxemic respiratory failure, 131 patients (age range, 1 month to 18 years) were prospectively followed. Parametric models were used to describe time-related events, and competing risks analysis was performed for mortality estimates. Multiple logistic analysis was applied to describe time-related predictors of ventilation time and mortality. Overall mortality was 27%. Peak oxygenation index (OI) measured at any time point (p < 0.001, 91% reliability in bootstrapping, after inverse transformation) and Pediatric Risk of Mortality, or PRISM, score within the first 12 hours of mechanical ventilation (p < 0.001, 63% reliability in bootstrapping, after square transformation) were identified as independent predictors of mortality. Peak OI, younger age, and need for renal replacement therapy were significantly associated with a longer time to extubation. Although OI was less reliable as outcome predictor within the first 12 hours of intubation, it still predicted duration of mechanical ventilation. No clear-cut threshold of OI was identified that could accurately predict mortality. Survival was characterized by a peak rate of extubations at approximately 1 week, with a more gradual decline thereafter, whereas death appeared as a constant risk over time, which exceeded chances of survival at approximately 4 weeks. Severity of oxygenation failure at any point in time during acute hypoxemic respiratory failure correlates with duration of mechanical ventilation and mortality. This is best reflected by the OI, which shows a direct correlation to outcome in a time independent manner. PMID- 15817804 TI - The end-of-life family conference: communication empowers. PMID- 15817803 TI - Lung volume and continuous positive airway pressure requirements in obstructive sleep apnea. AB - Previous studies have demonstrated that lung volume during wakefulness influences upper airway size and resistance, particularly in patients with sleep apnea. We sought to determine the influence of lung volume on the level of continuous positive airway pressure (CPAP) required to prevent flow limitation during non REM sleep in subjects with sleep apnea. Seventeen subjects (apnea-hypopnea index, 42.6 +/- 6.2 [SEM]) were studied during stable non-REM sleep in a rigid head-out shell equipped with a positive/negative pressure attachment for manipulation of extrathoracic pressure. An epiglottic pressure catheter plus a mask/pneumotachometer were used to assess flow limitation. When lung volume was increased by 1,035 +/- 22 ml, the CPAP level could be decreased from 11.9 +/- 0.7 to 4.8 +/- 0.7 cm H(2)O (p < 0.001) without flow limitation. The decreased CPAP at the same negative extrathoracic pressure yielded a final lung volume increase of 421 +/- 36 ml above the initial value. Conversely, when lung volume was reduced by 732 +/- 74 ml (n = 8), the CPAP level had to be increased from 11.9 +/ 0.7 to 17.1 +/- 1.0 cm H(2)O (p < 0.001) to prevent flow limitation, with a final lung volume decrease of 567 +/- 78 ml. These results demonstrate that relatively small changes in lung volume have an important effect on the upper airway in subjects with sleep apnea during non-REM sleep. PMID- 15817805 TI - From quantitative trait locus to gene: a work in progress. PMID- 15817807 TI - Influenza vaccination in children with asthma: no reason to change current recommendations. PMID- 15817806 TI - ATS/ERS recommendations for standardized procedures for the online and offline measurement of exhaled lower respiratory nitric oxide and nasal nitric oxide, 2005. PMID- 15817809 TI - Effects of pancreatic polypeptide on pancreas-projecting rat dorsal motor nucleus of the vagus neurons. AB - We investigated the pre- and postsynaptic effects of pancreatic polypeptide (PP) on identified pancreas-projecting neurons of the rat dorsal motor nucleus of the vagus in thin brain stem slices. Perfusion with PP induced a TTX- and apamin sensitive, concentration-dependent outward (22% of neurons) or inward current (21% of neurons) that was accompanied by a decrease in input resistance; PP was also found to affect the amplitude of the action potential afterhyperpolarization. The remaining 57% of neurons were unaffected. PP induced a concentration-dependent inhibition in amplitude of excitatory (n = 22 of 30 neurons) and inhibitory (n = 13 of 17 neurons) postsynaptic currents evoked by electrical stimulation of the adjacent nucleus of the solitary tract, with an estimated EC(50) of 30 nM for both. The inhibition was accompanied by an alteration in the paired pulse ratio, suggesting a presynaptic site of action. PP also decreased the frequency, but not amplitude, of spontaneous excitatory (n = 6 of 11 neurons) and inhibitory currents (n = 7 of 9 neurons). In five neurons, chemical stimulation of the area postrema (AP) induced a TTX-sensitive inward (n = 3) or biphasic (outward and inward) current (n = 2). Superfusion with PP reversibly reduced the amplitude of these chemically stimulated currents. Regardless of the PP-induced effect, the vast majority of responsive neurons had a multipolar somata morphology with dendrites projecting to areas other than the fourth ventricle or the central canal. These results suggest that pancreas projecting rat dorsal motor nucleus of the vagus neurons are heterogeneous with respect to their response to PP, which may underlie functional differences in the vagal modulation of pancreatic functions. PMID- 15817810 TI - A 5-HT4 agonist, mosapride, enhances intrinsic rectorectal and rectoanal reflexes after removal of extrinsic nerves in guinea pigs. AB - Distension-evoked reflex of rectorectal (R-R) contractions and rectointernal anal sphincter (R-IAS) relaxations can be generated in guinea pigs through an extrinsic sacral excitatory neural pathway (pelvic nerves) as well as intrinsic cholinergic excitatory and nitrergic inhibitory pathways. The aim of the present study was to create intrinsic R-R and R-IAS reflex models by pithing (destruction of the lumbar and sacral cords; PITH) and to evaluate whether the prokinetic benzamide mosapride, a 5-HT(4) receptor agonist, enhances these reflexes. The mechanical activities of the R-R and R-IAS were recorded in the anesthetized guinea pig on days 2-9 after PITH. Although the basal rectal pressure at distension after PITH was significantly lower than control, the reflex indexes of R-R contractions and synchronous R-IAS relaxations were unchanged between days 4 and 9 after PITH. The frequency of spontaneous rectal and IAS motility were also unchanged. Immunohistochemical studies revealed that the distribution of myenteric and intramuscular interstitial cells of Cajal (ICC) were not altered after PITH. Mosapride (0.1-1.0 mg/kg iv) dose-dependently increased both intrinsic R-R (maximum: 1.82) and R-IAS reflex indexes (maximum: 2.76) from control (1.0) 6-9 days after PITH. The 5-HT(4) receptor antagonist, GR-113808 (1.0 mg/kg iv) decreased the R-R and R-IAS reflex indexes by approximately 50% and antagonized the effect of mosapride (1.0 mg/kg iv). The present results indicate that mosapride moderately enhanced intrinsic R-R and R-IAS reflexes functionally compensated after deprivation of extrinsic nerves, mediated through endogenously active intrinsic 5-HT(4) receptors. PMID- 15817811 TI - Acute hepatic steatosis in mice by blocking beta-oxidation does not reduce insulin sensitivity of very-low-density lipoprotein production. AB - Accumulation of triglycerides (TG) in the liver is generally associated with hepatic insulin resistance. We questioned whether acute hepatic steatosis induced by pharmacological blockade of beta-oxidation affects hepatic insulin sensitivity, i.e., insulin-mediated suppression of VLDL production and insulin induced activation of phosphatidylinositol 3-kinase (PI3-kinase) and PKB. Tetradecylglycidic acid (TDGA), an inhibitor of carnitine palmitoyl transferase-1 (CPT1), was used for this purpose. Male C57BL/6J mice received 30 mg/kg TDGA or its solvent intraperitoneally and were subsequently fasted for 12 h. CPT1 inhibition resulted in severe microvesicular hepatic steatosis (19.9 +/- 8.3 vs. 112.4 +/- 25.2 nmol TG/mg liver, control vs. treated, P < 0.05) with elevated plasma nonesterified fatty acid (0.68 +/- 0.25 vs. 1.21 +/- 0.41 mM, P < 0.05) and plasma TG (0.39 +/- 0.16 vs. 0.60 +/- 0.10 mM, P < 0.05) concentrations. VLDL TG production rate was not affected on CPT1 inhibition (74.9 +/- 15.2 vs. 79.1 +/ 12.8 mumol TG.kg(-1).min(-1), control vs. treated) although treated mice secreted larger VLDL particles (59.3 +/- 3.6 vs. 66.6 +/- 4.5 nm diameter, P < 0.05). Infusion of insulin under euglycemic conditions suppressed VLDL production rate in control and treated mice by 43 and 54%, respectively, with formation of smaller VLDL particles (51.2 +/- 2.5 and 53.2 +/- 2.8 nm diameter). Insulin induced insulin receptor substrate (IRS)1- and IRS2-associated PI3-kinase activity and PKB-phosphorylation were not affected on TDGA treatment. In conclusion, acute hepatic steatosis caused by pharmacological inhibition of beta oxidation is not associated with reduced hepatic insulin sensitivity, indicating that hepatocellular fat content per se is not causally related to insulin resistance. PMID- 15817812 TI - Bile acid signaling through FXR induces intracellular adhesion molecule-1 expression in mouse liver and human hepatocytes. AB - Previous studies have demonstrated a dramatic induction of inflammatory gene expression in livers from mice fed a high-fat, high-cholesterol diet containing cholate after 3-5 wk. To determine the contribution of cholate in mediating these inductions, C57BL/6 mice were fed a chow diet supplemented with increasing concentrations of cholic acid (CA) for 5 days. A dose-dependent induction in the hepatic levels of TNF-alpha, VCAM-1, ICAM-1, and SAA-2 mRNA were observed. As positive controls, a dose-dependent repression of cholesterol 7alpha-hydroxylase and a dose-dependent induction of small heterodimer partner (SHP) expression were also observed, suggesting that farnesoid X receptor (FXR) was activated. In addition, ICAM-1 and SHP mRNA levels were also induced in primary human hepatocytes when treated with chenodeoxycholic acid or GW4064, a FXR-selective agonist. The involvement of FXR in CA-induced inflammatory gene expression was further investigated in the human hepatic cell line HepG2. Both ICAM-1 and SHP expression were induced in a dose- and time-dependent manner by treatment with the FXR-selective agonist GW4064. Moreover, the induction of ICAM-1 by GW4064 was inhibited by the FXR antagonist guggulsterone or with transfection of FXR siRNA. Finally, the activity of FXR was mapped to a retinoic acid response element (RARE) site containing an imbedded farnesoid X response element (FXRE) on the human ICAM-1 promoter and FXR and retinoid X receptor were demonstrated to bind to this site. Finally, FXR-mediated activation of ICAM-1 could be further enhanced by TNF-alpha cotreatment in hepatocytes, suggesting a potential cooperation between cytokine and bile acid-signaling pathways during hepatic inflammatory events. PMID- 15817813 TI - Kinesin and dynein move a peroxisome in vivo: a tug-of-war or coordinated movement? AB - We used fluorescence imaging with one nanometer accuracy (FIONA) to analyze organelle movement by conventional kinesin and cytoplasmic dynein in a cell. We located a green fluorescence protein (GFP)-tagged peroxisome in cultured Drosophila S2 cells to within 1.5 nanometers in 1.1 milliseconds, a 400-fold improvement in temporal resolution, sufficient to determine the average step size to be approximately 8 nanometers for both dynein and kinesin. Furthermore, we found that dynein and kinesin do not work against each other in vivo during peroxisome transport. Rather, multiple kinesins or multiple dyneins work together, producing up to 10 times the in vitro speed. PMID- 15817814 TI - Functional genomic analysis of the Wnt-wingless signaling pathway. AB - The Wnt-Wingless (Wg) pathway is one of a core set of evolutionarily conserved signaling pathways that regulates many aspects of metazoan development. Aberrant Wnt signaling has been linked to human disease. In the present study, we used a genomewide RNA interference (RNAi) screen in Drosophila cells to screen for regulators of the Wnt pathway. We identified 238 potential regulators, which include known pathway components, genes with functions not previously linked to this pathway, and genes with no previously assigned functions. Reciprocal-Best Blast analyses reveal that 50% of the genes identified in the screen have human orthologs, of which approximately 18% are associated with human disease. Functional assays of selected genes from the cell-based screen in Drosophila, mammalian cells, and zebrafish embryos demonstrated that these genes have evolutionarily conserved functions in Wnt signaling. High-throughput RNAi screens in cultured cells, followed by functional analyses in model organisms, prove to be a rapid means of identifying regulators of signaling pathways implicated in development and disease. PMID- 15817815 TI - High-resolution NMR spectroscopy with a portable single-sided sensor. AB - We report construction of a portable nuclear magnetic resonance sensor with a single-sided open probe design. The resulting magnetic field inhomogeneity is compensated by a pulse sequence that takes advantage of parallel inhomogeneity in the applied radio frequency field. We can thereby acquire fluorine-19 spectra of liquid fluorocarbons with 8 parts per million resolution, surmounting the long standing obstacle of obtaining chemical shift information with open probe instruments. PMID- 15817816 TI - A hydrogen-rich early Earth atmosphere. AB - We show that the escape of hydrogen from early Earth's atmosphere likely occurred at rates slower by two orders of magnitude than previously thought. The balance between slow hydrogen escape and volcanic outgassing could have maintained a hydrogen mixing ratio of more than 30%. The production of prebiotic organic compounds in such an atmosphere would have been more efficient than either exogenous delivery or synthesis in hydrothermal systems. The organic soup in the oceans and ponds on early Earth would have been a more favorable place for the origin of life than previously thought. PMID- 15817817 TI - Analysis of minor hemoglobins by matrix-assisted laser desorption/ionization time of-flight mass spectrometry. AB - BACKGROUND: Hemoglobin (Hb) heterogeneity arises mainly from posttranslational modifications of the globin chains, and cation-exchange chromatography reveals falsely increased concentrations of some minor Hbs in the presence of abnormal Hbs. Here we describe a method for identification of the globin chains and their posttranslational modifications contained in the Hb fractions. METHODS: We used cation-exchange HPLC (PolyCAT A column) for separation of Hb fractions and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for analysis of the separated globin chains. Globin chains were identified by their molecular masses. Posttranslational modifications of globin chains were identified by digestion of the proteins with endoproteinase V8 before MALDI-TOF MS of the resulting peptides. RESULTS: Analysis of the HbA2 fractions of patients with HbS revealed 4 different globin chains. We found, in addition to the expected alpha- and delta-chains, the carbamylated alpha- and the betaS-chains. Additionally, we analyzed HbH, Hb Barts, HbA 1b, pre-HbA 1c, HbA 1c, HbF1, HbF, HbA 1d3a, HbA 1d3b, HbA2, and HbC1 fractions from control and pathologic blood samples. We identified several posttranslational modifications of the globin chains, such as pyruvatization, glycation, acetylation, carbamylation, and acetaldehyde adduct formation. CONCLUSIONS: The native and posttranslationally modified globin chains in minor and major Hbs are unambiguously identified by MALDI-TOF MS. A minor Hb containing the carbamylated alpha- and the betaS-chain elutes at the same time as normal HbA2 (alpha2delta2) and thus leads to falsely increased HbA2 values in patients with HbS when blood is analyzed with PolyCAT A chromatography. PMID- 15817818 TI - Monitoring of clopidogrel action: comparison of methods. AB - BACKGROUND: Clopidogrel is a potent drug for prevention of adverse effects during and after coronary intervention. Increasing experience indicates that a significant proportion of patients do not respond adequately to clopidogrel. Because failure of antiplatelet therapy can have severe consequences, there is need for a reliable assay to quantify the effectiveness of clopidogrel treatment. METHODS: Of 24 healthy volunteers admitted to the study, 18 were treated for 1 week with clopidogrel (300-mg loading dose and 75-mg maintenance dose), and 6 with placebo. Platelet function was monitored by 2 assays, based on flow cytometry and enzyme immunoassay, that measure the phosphorylation status of vasodilator-stimulated phosphoprotein (VASP) and by aggregometry, flow cytometry of P-selectin, and the platelet function analyzer at baseline, on days 1-5, and on day 9 of treatment. RESULTS: Aggregometry and VASP phosphorylation revealed a loss of platelet response to ADP within 12 h after clopidogrel intake. The phosphorylation status of VASP correlated with the inhibition of platelet aggregation. In contrast, neither P-selectin expression nor PFA-100 closure time was a clear indicator of clopidogrel effects on platelets. CONCLUSIONS: VASP phosphorylation assays are reliable for quantifying clopidogrel effects. Because the VASP assay directly measures the function of the clopidogrel target, the P2Y12 receptor, the assay is selective for clopidogrel effects rather than effects of other platelet inhibitors commonly in use. PMID- 15817819 TI - Identification of subtypes of CYP2D gene rearrangements among carriers of CYP2D6 gene deletion and duplication. AB - BACKGROUND: Cytochrome P450 2D6 (CYP2D6) is one of the best-known polymorphic drug-metabolizing enzymes. Rapidly evolving genotyping techniques permit the identification of single-nucleotide polymorphisms (SNPs) and thereby a prediction of individual metabolic capacities for CYP2D6 substrates. A considerable part of interindividual variability in CYP2D6 enzyme activity, however, is not related to SNPs but to gene deletions and duplications. Currently used genotyping methods assume that these gene rearrangements are homogeneous. METHODS: We analyzed the interindividual variability in CYP2D6 gene arrangements in genomic DNA from 740 Caucasian individuals by allele-specific PCR to identify common SNPs of the CYP2D6 gene that correspond to the variant alleles CYP2D6*3, *4, and *9. We investigated the presence and variability of CYP2D6*5 (gene deletion), CYP2D6x2 (gene duplication), and CYP2D6xn (gene amplification) by EcoRI and XbaI restriction fragment length polymorphism analyses and by long PCR plus KpnI and BamHI digestion. The presence of new mutations at the CYP2D locus was analyzed by sequencing. RESULTS: CYP2D6 gene rearrangements were present in >12% of individuals. Variability in the rearrangements regarding both gene deletion and gene duplication existed, and one of the unusual arrangements led to incorrect phenotype prediction. The frequency for carriers of unusual gene rearrangements was <0.3% (95% confidence interval, 0%-0.6%) in the population studied. CONCLUSIONS: Heterogeneity in CYP2D6 gene rearrangement exists, but the allele frequency indicates that the risk for an erroneous phenotype prediction related to such variability is extremely low and that this risk can be neglected in routine analyses. PMID- 15817821 TI - Evaluation of a new CA15-3 protein assay method: optical protein-chip system for clinical application. PMID- 15817820 TI - Multimarker quantitative real-time PCR detection of circulating melanoma cells in peripheral blood: relation to disease stage in melanoma patients. AB - BACKGROUND: Detection of melanoma cells in circulation may be important in assessing tumor progression. The objective of this study was to develop a specific, reliable multimarker quantitative real-time reverse transcription-PCR (qRT) assay for detecting melanoma cells in patients' blood. METHODS: We developed qRT assays for the mRNA of four melanoma-associated markers: MART-1, GalNAc-T, PAX-3, and MAGE-A3. In optimization studies, we tested 17 melanoma cell lines and 49 peripheral blood leukocyte (PBL) samples from volunteers. We performed RNA and melanoma cell dilution studies to assess the detection limits and imprecision of the assays. We measured the mRNAs in blood specimens from 94 melanoma patients [American Joint Committee on Cancer (AJCC) stage I, n = 20; II, n = 20; III, n = 32; IV, n = 22]. RESULTS: All markers were frequently detected in melanoma cell lines, whereas none of the markers was detected in PBLs from volunteers. The qRT assay could detect 1 melanoma cell in 10(7) PBLs in the melanoma cell-dilution studies. Markers were detected in 15%, 30%, 75%, and 86% of melanoma patients with AJCC stage I, II, III, and IV disease, respectively. The number of positive markers and AJCC stage were significantly correlated (Spearman correlation coefficient = 0.58; P < 0.0001). CONCLUSIONS: Multimarker qRT can detect circulating melanoma cells in blood. Measurement of the studied molecular markers in blood may be useful in detection of metastasis and monitoring treatment response of melanoma patients. PMID- 15817822 TI - Role of the RIP140 corepressor in ovulation and adipose biology. AB - RIP140 is a ligand-dependent corepressor for most, if not all, nuclear receptors. It is expressed widely in many different tissues, but the phenotype of mice devoid of RIP140 indicates that it plays a crucial role in the ovary and in adipose biology. Ovarian expression of RIP140 is cell-type-specific during follicular development and it is essential for oocyte release during ovulation, but not for luteinization of mature ovarian follicles. In adipose tissue, RIP140 is essential for normal fat accumulation and RIP140-null mice show decreased lipid storage even on a high-fat diet, with upregulation of mitochondrial uncoupling protein (UCP1) in some fat depots. Thus RIP140 plays a crucial role in female fertility and in energy homeostasis, and could be a target for infertility treatment, new contraceptive strategies or prevention of obesity. PMID- 15817823 TI - Interdependence of oestrogen and insulin-like growth factor-I in the brain: potential for analysing neuroprotective mechanisms. AB - The actions of oestradiol in the brain involve interaction with growth factors, such as insulin-like growth factor-I (IGF-I). Many cells in the brain co-express receptors for oestradiol and IGF-I and both factors interact to regulate neural function. The relationship of oestrogen receptor alpha with IGF-I receptor through the mitogen-activated protein kinase and the phosphoinositide 3-kinase signalling pathways may represent the point of convergence used by these two factors to cooperatively modulate neuritic growth, synaptic plasticity, neuroendocrine events, reproductive behaviour and neuronal survival. In addition, Akt and glycogen synthase kinase 3beta are key molecular targets to explain the interaction of oestrogen and IGF-I receptor signalling in the promotion of neuroprotection. PMID- 15817824 TI - Protein tyrosine phosphatases and signalling. AB - A cornerstone of many cell-signalling events rests on reversible phosphorylation of tyrosine residues on proteins. The reversibility relies on the coordinated actions of protein tyrosine kinases and protein tyrosine phosphatases (PTPs), both of which exist as large protein families. This review focuses on the rapidly evolving field of the PTPs. We now know that rather than simply scavenging phosphotyrosine, the PTPs specifically regulate a wide range of signalling pathways. To illustrate this and to highlight current areas of agreement and contention in the field, this review will present our understanding of PTP action in selected areas and will present current knowledge surrounding the regulatory mechanisms that control PTP enzymes themselves. It will be seen that PTPs control diverse processes such as focal adhesion dynamics, cell-cell adhesion and insulin signalling, and their own actions are in turn regulated by dimerisation, phosphorylation and reversible oxidation. PMID- 15817825 TI - Substitution of the cysteine 438 residue in the cytoplasmic tail of the glucagon like peptide-1 receptor alters signal transduction activity. AB - Several G-protein-coupled receptors contain cysteine residues in the C-terminal tail that may modulate receptor function. In this work we analysed the substitution of Cys438 by alanine in the glucagon-like peptide-1 (GLP-1) receptor (GLPR), which led to a threefold decrease in cAMP production, although endocytosis and cellular redistribution of GLP-1 receptor agonist-induced processes were unaffected. Additionally, cysteine residues in the C-terminal tail of several G-protein-coupled receptors were found to act as substrates for palmitoylation, which might modify the access of protein kinases to this region. His-tagged GLP-1 receptors incorporated 3H-palmitate. Nevertheless, substitution of Cys438 prevented the incorporation of palmitate. Accordingly, we also investigated the effect of substitution of the consensus sequence by protein kinase C (PKC) Ser431/432 in both wild-type and Ala438 GLP-1 receptors. Substitution of Ser431/432 by alanine did not modify the ability of wild-type receptors to stimulate adenylate cyclase or endocytosis and recycling processes. By contrast, the substitution of Ser431/432 by alanine in the receptor containing Ala438 increased the ability to stimulate adenylate cyclase. All types of receptors were mainly internalised through coated pits. Thus, cysteine 438 in the cytoplasmic tail of the GLP-1 receptor would regulate its interaction with G proteins and the stimulation of adenylyl cyclase. Palmitoylation of this residue might control the access of PKC to Ser431/432. PMID- 15817826 TI - Growth factor-induced signaling of the pancreatic epithelium. AB - Activated signaling proteins regulate diverse processes, including the differentiation of the pancreatic islet cells during ontogeny. Here we uncover the in vivo phosphorylation status of major growth factor-activated signaling proteins in normal adult mice and during pancreatic islet regeneration. We report elevated phospho-mitogen-activated protein kinase (phospho-MAPK), phospho-c-Jun NH2-terminal kinase (phospho-JNK), and phospho-p38 MAPK expression during pancreatic regeneration. Immunoblotting experiments demonstrated elevated phosphorylation of p52 Src-homology/collagen (SHC) in the ductal network as well, substantiating the activation of this pathway. Furthermore, protein kinase B (PKB/Akt), a key signaling protein in the anti-apoptotic pathway, was phosphorylated to a greater extent in the ductal network from regenerating pancreas. We observed fibroblast growht factor (FGF)10 and platelet-derived growth factor (PDGF)AA expression in embryonic as well as regenerating adult pancreas. Epidermal growth factor (EGF) and PDGFAA stimulated MAPK and Akt phosphorylation, while FGF10 stimulated MAPK but not Akt phosphorylation in a time-dependent manner in freshly isolated cells from the adult ductal network. These data suggest that a heightened level of expression and stimulation of key signaling proteins underlie the expansion and differentiation processes that support pancreatic ontogeny and regeneration. PMID- 15817827 TI - Intragranular targeting of syncollin, but not a syncollinGFP chimera, inhibits regulated insulin exocytosis in pancreatic beta-cells. AB - Several proteins play a role in the mechanism of insulin exocytosis. However, these 'exocytotic proteins' have yet to account for the regulated aspect of insulin exocytosis, and other factors are involved. In pancreatic exocrine cells, the intralumenal zymogen granule protein, syncollin, is required for efficient regulated exocytosis, but it is not known whether intragranular peptides similarly influence regulated insulin exocytosis. Here, this issue has been addressed using expression of syncollin and a syncollin-green fluorescent protein (syncollinGFP) chimera in rat islet beta-cells as experimental tools. Syncollin is not normally expressed in beta-cells but adenoviral-mediated expression of both syncollin and syncollinGFP indicated that these were specifically targeted to the lumen of beta-granules. Syncollin expression in isolated rat islets had no effect on basal insulin secretion but significantly inhibited regulated insulin secretion stimulated by glucose (16.7 mM), glucagon-like peptide-1 (GLP-1) (10 nM) and glyburide (5 microM). Consistent with specific localization of syncollin to beta-granules, constitutive secretion was unchanged by syncollin expression in rat islets. Syncollin-mediated inhibition of insulin secretion was not due to inadequate insulin production. Moreover, secretagogue-induced increases in cytosolic intracellular Ca2+, which is a prerequisite for triggering insulin exocytosis, were unaffected in syncollin-expressing islets. Therefore, syncollin was most likely acting downstream of secondary signals at the level of insulin exocytosis. Thus, syncollin expression in beta-cells has highlighted the importance of intralumenal beta-granule peptide factors playing a role in the control of insulin exocytosis. In contrast to syncollin, syncollinGFP had no effect on insulin secretion, underlining its usefulness as a 'fluorescent tag' to track beta-granule transport and exocytosis in real time. PMID- 15817828 TI - External ATP triggers Ca2+ signals suited for synchronization of pancreatic beta cells. AB - External ATP is supposed to trigger short-lived increases (transients) of cytoplasmic Ca2+ important for entraining insulin-secreting beta-cells into a common rhythm. To get insight into this process, rises of the cytoplasmic Ca2+ concentration ([Ca2+]i) induced by external ATP were compared with those obtained with acetylcholine, another neurotransmitter with stimulatory effects on the inositol trisphosphate (IP3) production. A ratiometric fura-2 technique was used for measuring [Ca2+]i in individual beta-cells and small aggregates isolated from ob/ob mouse islets and superfused with a medium containing methoxyverapamil. ATP and acetylcholine induced temporary rises of [Ca2+]I from a basal level manifested as solitary transients (<20 s) and bumps (> or =20 s) superimposed or not with transients. Addition of ATP (1-100 microM) usually triggered transients whereas acetylcholine induced bumps lacking superimposed transients. After the initial rise there was a steady-state elevation of [Ca2+]i in beta-cells exposed to acetylcholine but not to ATP. Similar differences were seen comparing the responses of rat beta-cells to 100 microM ATP and acetylcholine. Inhibition of the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) pump (with 50 microM cyclopiazonic acid) prevented both the ATP-induced rise of [Ca2+]i and the spontaneous firing of transients. Similar effects were seen after activation of protein kinase C (10 nM phorbol-12-myristate-13-acetate), whereas an inhibitor of this enzyme (2 microM bisindolylmaleimide) promoted the generation of transients. The results indicate that ATP fulfils the demands for a coordinator of the secretory activity of beta-cells by generating distinct [Ca2+]i transients without sustained elevation of basal [Ca2+]i. PMID- 15817829 TI - 1,25-dihydroxyvitamin D3 stimulates vesicular transport within 5 s in polarized intestinal epithelial cells. AB - Controversy remains regarding whether the seco-steroid hormone 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) enhances calcium and phosphate movement across the intestinal epithelial cell by facilitated diffusion or a vesicular transport mechanism. In this study we investigated whether membrane trafficking, as judged by confocal microscopy, was sufficiently rapid in comparison to hormone stimulated uptake of phosphate (32P). Primary cultures of chick intestinal cells were established overnight either in Petri dishes (uptake studies) or chambered coverslips (confocal microscopy). Addition of 130 pM 1,25(OH)2D3 resulted in an apparent increase in 32P uptake within 1 min, relative to controls, that was statistically significant from 3-10 min of incubation. Using the endocytic marker dye, FM1-43, confocal microscopy revealed a profound decrease in membrane associated fluorescence (apical> basal) within 10 s of hormone treatment, a return of fluorescence at 15-65 s, followed by another round of decreasing and increasing fluorescence. Between 3-9 min of incubation, fluorescence intensity increased 50% (apical region) and 20% (basal region) over control conditions. An antibody (Ab 099) directed against a putative membrane receptor for 1,25(OH)2D3 (1,25D3-MARRS) inhibited both 32P uptake, and changes in fluorescence. In addition, the protein kinase C (PKC) inhibitor, calphostin C, inhibited both 32P uptake and the observed 1,25(OH)2D3-mediated changes in fluorescence. At the microscopic level, calphostin C pretreatment abolished the very rapid redistribution of the endocytic marker dye, although a slight increase in fluorescence was still observed. We conclude that 1,25(OH)2D3-stimulated vesicular trafficking is mediated by the 1,25D3-MARRS protein, implicates a PKC signaling mechanism, and occurs in a time frame that is commensurate with a role in ion transport. PMID- 15817830 TI - Thyroid hormone export from cells: contribution of P-glycoprotein. AB - Verapamil inhibits tri-iodothyronine (T3) efflux from several cell types, suggesting the involvement of multidrug resistance-associated (MDR) proteins in T3 transport. The direct involvement of P-glycoprotein (P-gp) has not, however, been investigated. We compared the transport of 125I-T3 in MDCKII cells that had been transfected with mdr1 cDNA (MDCKII-MDR) versus wild-type MDCKII cells (MDCKII), and examined the effect of conventional (verapamil and nitrendipine) and specific MDR inhibitors (VX 853 and VX 710) on 125I-T3 efflux. We confirmed by Western blotting the enhanced expression of P-gp in MDCKII-MDR cells. The calculated rate of 125I-T3 efflux from MDCKII-MDR cells (around 0.30/min) was increased twofold compared with MDCKII cells (around 0.15/min). Overall, cellular accumulation of 125I-T3 was reduced by 26% in MDCKII-MDR cells compared with MDCKII cells, probably reflecting enhanced export of T3 from MDCKII-MDR cells rather than reduced cellular uptake, as P-gp typically exports substances from cells. Verapamil lowered the rate of 125I-T3 efflux from both MDCKII and MDCKII MDR cells by 42% and 66% respectively, while nitrendipine reduced 125I-T3 efflux rate by 36% and 48% respectively, suggesting that both substances inhibited other cellular T3 transporters in addition to P-gp. The specific MDR inhibitors VX 853 and VX 710 had no effect of 125I-T3 efflux rate from wild-type MDCKII cells but reduced 125I-T3 export in MDCKII-MDR cells by 50% and 53% respectively. These results have provided the first direct evidence that P-gp exports thyroid hormone from cells. PMID- 15817831 TI - Reciprocal regulation of activin A and inhibin B by interleukin-1 (IL-1) and follicle-stimulating hormone (FSH) in rat Sertoli cells in vitro. AB - In several biological systems, the inhibin beta(A) homodimer activin A is stimulated by, and in turn, inhibits the action of interleukin (IL)-1 (both IL 1alpha and IL-1beta) and IL-6. The possibility that a similar regulatory relationship operates within the testis was investigated. Sertoli cells from immature (20-day-old) rats were cultured with human IL-1alpha or IL-1beta, human IL-6 and/or ovine FSH or dibutyryl cAMP. Activin A and the inhibin dimers, inhibin A and inhibin B, were measured by specific ELISA. Immunoreactive inhibin (ir-inhibin) was measured by RIA. Activin/inhibin subunit mRNA expression was measured by quantitative real-time PCR. Both IL-1 isoforms, but not IL-6, stimulated activin A secretion through increased synthesis of beta(A)-subunit mRNA. IL-1 also stimulated activin A secretion by testicular peritubular cells. In contrast to the effect on activin A, IL-1 suppressed inhibin beta(B)-subunit and, to a lesser extent, alpha-subunit mRNA expression, thereby reducing basal and FSH-stimulated inhibin B secretion by the Sertoli cells. Conversely, FSH inhibited basal activin A secretion and antagonised the stimulatory effects of IL 1. Dibutyryl cAMP partially inhibited the action of IL-1 on activin A secretion, but had no significant effect on basal activin A secretion. Secretion of inhibin A was low in all treatment groups. These data demonstrate that IL-1 and FSH/cAMP exert a reciprocal regulation of activin A and inhibin B synthesis and release by the Sertoli cell, and suggest a role for activin A as a potential feedback regulator of IL-1 and IL-6 activity in the testis during normal spermatogenesis and in inflammation. PMID- 15817832 TI - Thyroid status and nitric oxide in rat arterial vessels. AB - Thyroid disease has profound effects on cardiovascular function. Hypo- and hyperthyroidism, for example, are associated with reduced and increased maximal endothelium-dependent vasodilation respectively. We therefore hypothesized that the capacity for vascular nitric oxide (NO) formation is decreased in hypothyroidism and increased in hyperthyroidism. To test this hypothesis, rats were made hypothyroid (HYPO) with propylthiouracil or hyperthyroid (HYPER) with triiodothyronine over 3-4 months. Compared with euthyroid control rats (EUT), HYPO exhibited blunted growth and lower citrate synthase activity in the soleus muscle; HYPER exhibited left ventricular hypertrophy and higher citrate synthase activity in the soleus muscle (P<0.05 for all effects). The capacity for NO formation was determined in aortic extracts by formation of [3H]L-citrulline from [3H]L-arginine, i.e. NO synthase (NOS) activity. Thyroid status modulated NOS activity (EUT, 36.8 +/- 5.5 fmol/h per mg protein; HYPO, 26.0 +/- 7.9; HYPER, 64.6 +/- 12.7; P<0.05, HYPER vs HYPO). Expression of endothelial and neural isoforms of NOS was modulated by thyroid status in a parallel fashion. Capacity for responding to NO was also determined via measuring cGMP concentration in aortae incubated with sodium nitroprusside. Stimulated cGMP formation was also modulated by thyroid status (EUT, 73.0 +/- 20.2 pmol/mg protein; HYPO, 152.4 +/- 48.7; HYPER, 10.4 +/- 2.6; P<0.05, HYPER vs HYPO). These data indicate that thyroid status alters capacities for both formation of and responding to NO. The former finding may contribute to previous findings concerning vascular function in thyroid disease states. PMID- 15817833 TI - Nongenomic effect of thyroid hormone on free-radical production in human polymorphonuclear leukocytes. AB - Over the past few years increasing evidence has suggested the nongenomic effects of thyroid hormone, such as the activation of the signal transduction pathways and the activation of nuclear factor-kappaB by the induction of oxidative stress. The present study was undertaken to investigate the effect of thyroid hormone on human polymorphonuclear leukocytes (PMNLs) which are known as important sources of reactive oxygen species in the circulation. The production of superoxide anion (O2-) and the activity of myeloperoxidase were determined in the presence and absence of several inhibitors of the signalling pathway. L-thyroxine (T4) l 3,5,3'-tri-iodothyronine (T3) and L-3,5-di-iodothyronine (T2) stimulated O2- production in PMNLs in a dose-dependent manner within a few minutes of addition to cells. Thyroid hormone-stimulated O2- production was partially inhibited by pertussis toxin, an inhibitor of GTP-binding G protein, and was completely abolished by the protein kinase C inhibitors calphostin C and Ro-32-0432, and by a calcium chelator (BAPTA; bis-(o-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid). Thyroid hormone stimulated myeloperoxidase activity and induced 125I- incorporation into PMNLs. Furthermore, thyroid hormone pre-incubation enhanced O2 production for n-formyl-methionyl-leucyl- phenylalanine (FMLP) stimulation. In conclusion, novel nongenomic actions of thyroid hormone, the induction of superoxide anion production and the stimulation of myeloperoxidase activity in PMNLs were demonstrated. The induction of O2- production requires calcium and is mediated by a pertussis toxin-sensitive G protein via stimulation of protein kinase C(s). These results suggest the existence of a membrane-bound binding site for thyroid hormone in PMNLs and a physiological role for thyroid hormone in the cellular defence mechanisms by stimulating free-radical production. PMID- 15817834 TI - Dexamethasone suppresses Smad3 pathway in osteoblastic cells. AB - Central in the pathogenesis of glucocorticoid (GC)-induced osteoporosis is the effects of GC on bone formation. However, the mechanism of GC-inhibited bone formation is not well known. Transforming growth factor (TGF)-beta is most abundant in bone matrix compared with other tissues, and we have recently proposed that Smad3, a TGF-beta signaling molecule, is important for promoting bone formation. However, no reports have been available about the effects of GC on Smad3 in osteoblasts. In the present study, we investigated whether dexamethasone (Dex), an active GC analog, would affect the expression and activity of Smad3 in mouse osteoblastic MC3T3-E1 and rat osteoblastic UMR-106 cells. Dex significantly suppressed Smad3-stimulated alkaline phosphatase (ALP) activity, although it did not affect TGF-beta-inhibited ALP activity in MC3T3-E1 cells. Moreover, pretreatment with Dex suppressed TGF-beta-enhanced expression of type I collagen in MC3T3-E1 and UMR-106 cells. In the luciferase assay using p3TP Lux with a Smad3-specific response element, Dex significantly suppressed the transcriptional activity induced by TGF-beta as well as Smad3. However, Dex did not affect the expression of Smad3 in these cells at both mRNA and protein levels. In conclusion, the present study indicates that Dex inhibits ALP activity and type I collagen expression, presumably by suppressing Smad3-induced transcriptional activity but not by modulating Smad3 expression in osteoblastic cells. PMID- 15817835 TI - Effects of age and pregnancy on the circulatory activin response of sheep to acute inflammatory challenge by lipopolysaccharide. AB - The release of activin A in response to intravenous injection of the bacterial cell-wall component lipopolysaccharide (LPS) was investigated in an ovine model of acute inflammatory challenge in newborn and adult sheep, and in non-pregnant and pregnant ewes. Neonatal lambs (<20 days of age) showed a quantitatively similar response in terms of circulating concentrations of activin A, its binding protein follistatin and the cytokine interleukin-6 compared with adult ewes challenged with an equivalent dose (300 ng/kg bodyweight) of LPS. The fever response and plasma tumour necrosis factor-alpha release in response to LPS, however, were significantly (P < 0.01) less in lambs than in the adult group. Pregnant ewes in the last trimester of gestation had similar responses to LPS, in all aspects measured, compared with their non-pregnant counterparts, apart from an ablated fever response. Although the adult and neonatal sheep responded to LPS, a similar response was not apparent in the fetal circulation, possibly due to a protective effect of the placenta. A 10-fold increase in the dose of LPS (from 300 ng to 3 microg/kg bodyweight) given to neonatal lambs elicited an increase in several cytokine responses measured, with a significant (P< 0.05) increase in follistatin release. In contrast, the amount of activin released by the increased dose of LPS was similar to that invoked by the lower dose. The effect of tolerance to LPS was investigated by giving a second challenge of LPS 5 days after the initial injection. In all animals studied, there was an ablated (P < 0.05) response to the subsequent LPS injection, apart from a similar temperature-response profile. These data provide further evidence that activin A concentrations in the bloodstream are acutely responsive to inflammatory challenge in post-natal life and suggest that the response forms a significant component of the innate immune system. PMID- 15817836 TI - Transgenic overexpression of neuromedin U promotes leanness and hypophagia in mice. AB - Recent work has shown that neuromedin U (NmU), a peptide initially identified as a smooth muscle contractor, may play a role in regulating food intake and energy homeostasis. To further evaluate this putative function, we measured food intake, body weight, energy expenditure and glucose homeostasis in transgenic mice that ubiquitously overexpress murine proNmU. NmU transgenic mice were lighter and had less somatic and liver fat, were hypophagic, and had improved insulin sensitivity as judged by an intraperitoneal insulin tolerance test. Transgenic mice had higher levels of hypothalamic NPY, POMC and MCH mRNA. There was no difference in O2 consumption between genotypes; however, NmU transgenic mice displayed a modest increase in respiratory quotient during food deprivation and refeeding. There were no behavioral disturbances in the NmU transgenic mice that could account for the results (e.g. changes in locomotor activity). When placed on a high-fat diet, transgenic mice remained lighter than wild-type mice and ate less, but gained weight at a rate similar to wild-type mice. Despite the increased weight gain with high-fat feeding, glucose tolerance was significantly improved in the transgenic mice. These findings support the hypothesized role of NmU as an endogenous anorexigenic peptide. PMID- 15817837 TI - Dehydroepiandrosterone (DHEA) modulates GHRH, somatostatin and angiotensin II action at the pituitary level. AB - In view of the present controversy related to the potential beneficial effects of clinical dehydroepiandrosterone (DHEA) treatments, and considering our own previous results that reveal an influence of this steroid in pituitary hyperplasia development in vivo in rats, we decided to evaluate the role of DHEA in prolactin and GH secretion, as well as in second messengers involved, in cultured rat anterior pituitary cells. DHEA (1 x 10(-5) to 1 x 10(-7) M) did not modify basal GH or prolactin release, and a prolactin inhibitory effect was observed only for androstenediol, a metabolite of DHEA. DHEA partially prevented dopamine (1 x 10(-6) M)-induced prolactin inhibition and facilitated the prolactin-releasing effect of 10(-8) M Ang II, without modifying the resulting Ca2+(i) mobilization. Furthermore, DHEA potentiated the GH release and cAMP production induced by 1 x 10(-8) M GHRH. Finally, DHEA partially reversed the inhibitory effect of 1 x 10(-8) M somatostatin on GH, but not prolactin, release. We conclude that DHEA in vitro, directly or indirectly through conversion into metabolites, is able to modulate the hormonal response of the pituitary to hypothalamic regulators. It can enhance pituitary prolactin release and induce GH secretion. These effects could help explain some of the side effects observed in prolonged DHEA treatments in vivo and should be taken into account when considering its use in human clinical trials. PMID- 15817838 TI - Pit-1beta reduces transcription and CREB-binding protein recruitment in a DNA context-dependent manner. AB - Many transcription factors are expressed as multiple isoforms with distinct effects on the regulation of gene expression, and the functional consequences of structural differences between transcription factor isoforms may allow for precise control of gene expression. The pituitary transcription factor isoforms Pit-1 and Pit-1beta differentially regulate anterior pituitary hormone gene expression. Pit-1 is required for the development of and appropriate hormone expression by anterior pituitary somatotrophs and lactotrophs. Pit-1beta differs structurally from Pit-1 by the splice-insertion of the 26-residue beta-domain in the trans-activation domain, and it differs functionally from Pit-1 in that it represses expression of the prolactin promoter in a cell-type specific manner. In order to identify signal and promoter context requirements for repression by Pit 1beta, we examined its function in the presence of physiological regulatory signals as well as wild-type and mutant Pit-1-dependent target promoters. Here, we demonstrate that Pit-1beta impairs recruitment of cAMP response element binding protein (CREB)-binding protein to the promoters that it represses. In addition, we show that repression of target promoter activity, reduction in promoter histone acetylation, and decrease of CREB-binding protein recruitment all depend on promoter context. These findings provide a mechanism for promoter specific repression by Pit-1beta. PMID- 15817839 TI - A steroidogenic cell line with differentiation potential from mouse granulosa cells, transfected with Ad4BP and SV40 large T antigen genes. AB - Several steroidogenic cell lines of granulosa cells (GC) have been used to elucidate differentiation mechanisms of GC during folliculogenesis. These cell lines, however, are of limited usefulness since they have lost some of their differentiation potential. The transcription factor adrenal-4 binding protein (Ad4BP), also known as steroidogenic factor-1 or NR5A1, is essential for the expression of all P-450 steroidogenic enzymes. By transfection with the Ad4BP gene together with SV40 DNA, we have generated several steroidogenic cell lines. One selective clone, named 4B2, retained its steroidogenic potential and was therefore analyzed in depth. This cell line responded to 8-Br-cAMP by displaying differentiation characteristics similar to those occurring in the differentiation process of primary cultured GC, including enhanced progesterone secretion, a cell shape change from a fibroblastic to epithelioid conformation, elongated mitochondria, increased gap junction formation and inhibition of cell proliferation. Prostaglandin E2 (PGE2), an intraovarian regulator of GC, stimulated cAMP production, and this eicosanoid, like 8-Br-cAMP, induced differentiation properties with the exception of cell conformation in 4B2 cells. These results suggest that expression of Ad4BP may provide the basis for a repertoire of cAMP-sensitive differentiation properties, including morphological alterations and growth inhibition. Thus, the 4B2 cell line may serve as a tool for elucidation of differentiation mechanisms that are under the control of Ad4BP. PMID- 15817840 TI - Production of recombinant porcine IGF-binding protein-5 and its effect on proliferation of porcine embryonic myoblast cultures in the presence and absence of IGF-I and Long-R3-IGF-I. AB - IGF-binding protein-5 (IGFBP-5) is produced by porcine embryonic myogenic cell (PEMC) cultures and is secreted into the medium. IGFBP-5 may play some role in myogenesis and/or in changes in myogenic cell proliferation that accompany differentiation. IGFBP-5 reportedly may either suppress or stimulate proliferation or differentiation of cultured cells depending on cell type and culture conditions. Additionally, IGFBP-5 has been shown to possess both IGF dependent and IGF-independent actions in some cell types. The goal of this study was to produce recombinant porcine IGFBP-5 (rpIGFBP-5) and assess its IGF-I dependent and IGF-I-independent actions on the proliferation of PEMCs. To accomplish this, we have expressed porcine IGFBP-5 in the baculovirus system, purified and characterized the expressed rpIGFBP-5 and produced an anti-porcine IGFBP-5 antibody that neutralizes the biological activity of porcine IGFBP-5. rpIGFBP-5, purified to 98% homogeneity using nickel affinity chromatography and IGF-I affinity chromatography, suppressed IGF-I-stimulated proliferation of PEMCs in a concentration-dependent manner (P>0.05). rpIGFBP-5 also suppressed Long-R3 IGF-I-stimulated proliferation of PEMCs (P>0.05), even in the presence of significant molar excess of Long-R3-IGF-I compared with rpIGFBP-5, demonstrating the IGF-independent activity that rpIGFBP-5 possesses in PEMCs, since Long-R3-IGF I is an IGF analog that has very low affinity for the IGFBPs but retains its ability to bind to the type I IGF receptor and thereby can stimulate proliferation. The anti-rpIGFBP-5 IgY produced against rpIGFBP-5 specifically recognized native porcine IGFBP-5 in PEMC media that also contained porcine IGFBP 2, -3, and -4. This antibody is capable of neutralizing the effects of both rpIGFBP-5 and endogenously produced porcine IGFBP-5 on PEMCs as well as detecting IGFBP-5 in Western blots. The production of rpIGFBP-5 and a neutralizing antibody to porcine IGFBP-5 provides a powerful tool to investigate the role of IGFBP-5 in porcine myogenic cell proliferation and differentiation. The data provided here demonstrated that IGFBP-5 has the potential to affect proliferation of PEMCs during critical periods of in vitro muscle cell development and therefore may impact the capacity for ultimate postnatal muscle mass development in vivo. PMID- 15817841 TI - Ghrelin is an orexigenic and metabolic signaling peptide in the arcuate and paraventricular nuclei. AB - Ghrelin is a 28-amino acid acylated peptide and is the endogenous ligand for the growth hormone secretagogue receptor (GHS-R). The GHS-R is expressed in hypothalamic nuclei, including the arcuate nucleus (Arc) where it is colocalized with neuropeptide Y (NPY) neurons. In the present study, we examined the effects of ghrelin on feeding and energy substrate utilization (respiratory quotient; RQ) following direct injections into either the arcuate or the paraventricular nucleus (PVN) of the hypothalamus. Ghrelin was administered at the beginning of the dark cycle at doses of 15-60 pmol to male and female rats. In feeding studies, food intake was measured 2 and 4 h postinjection. Separate groups of rats were injected with ghrelin, and the RQ (VCO(2)/VO(2)) was measured using an open circuit calorimeter over a 4-h period. Both Arc and PVN injections of ghrelin increased food intake in male and female rats. Ghrelin also increased RQ, reflecting a shift in energy substrate utilization in favor of carbohydrate oxidation. Because these effects are similar to those observed after PVN injection of NPY, we then assessed the impact of coinjecting ghrelin with NPY into the PVN. When rats were pretreated with very low doses of ghrelin (2.5-10 pmol), NPY's (50 pmol) effects on eating and RQ were potentiated. Overall, these data are in agreement with evidence suggesting that ghrelin functions as a gut brain endocrine hormone implicated in the regulation of food intake and energy metabolism. Our findings are also consistent with a possible interactive role of hypothalamic ghrelin and NPY systems. PMID- 15817843 TI - Mechanisms of insulin-dependent glucose transport into porcine and bovine skeletal muscle. AB - Euglycemic, hyperinsulinemic clamp tests have shown that adult ruminants are less insulin-sensitive than monogastric omnivores. The present study was carried out to elucidate possible cellular mechanisms contributing to this impaired insulin sensitivity of ruminants. Western blotting was used to measure glucose transporters 1 and 4 (GLUT1, GLUT4) in oxidative (musculus masseter and diaphragm) and glycolytic (musculus longissimus dorsi and semitendinosus) skeletal muscle in the crude membranes of pigs and cows. Muscles were characterized biochemically. To determine insulin-stimulated 3-O-D-[(3)H] methylglucose (3-O-MG) uptake and GLUT4 translocation, porcine and bovine musculus semitendinosus strips were removed by open muscle biopsy and incubated without and with 0.1 or 20 mIU insulin/ml. GLUT4 translocation was analyzed using subcellular fractionation techniques to isolate partially purified plasma membranes and cytoplasmic vesicles and using Western blotting. GLUT4 protein contents were significantly higher in oxidative than in glycolytic muscles in pigs and cows. GLUT1 protein contents were significantly higher in glycolytic than in oxidative muscles in bovines but not in porcines. The 3-O-MG uptake into musculus semitendinosus was similar in both species. Maximum insulin-induced GLUT4 translocation into musculus semitendinosus plasma membrane was significantly lower in bovines than in porcines. These results indicate that GLUT1 is the predominant glucose transporter in bovine glycolytic muscles and that a reinforced insulin-independent glucose uptake via GLUT1 may compensate for the impaired insulin-stimulated GLUT4 translocation, resulting in a similar 3-O MG uptake in bovine and porcine musculus semitendinosus. These findings may explain at least in part the impaired in vivo insulin sensitivity of adult ruminants compared with that of omnivorous monogastric animals. PMID- 15817844 TI - Racial differences in fat distribution: the importance of intermuscular fat. PMID- 15817845 TI - Isoflavones, protein, and bone. PMID- 15817846 TI - Vitamins E and C are safe across a broad range of intakes. AB - A robust database shows that dietary supplements of vitamins E and C are safe for the general population. Because these nutrients supply antioxidant and other functions for homeostasis and protection against free radical damage, supplementation has been intensively studied. Because of perceived benefits, many persons consume quantities of vitamins E and C well above the recommended dietary allowances. As safety guidance, tolerable upper intake levels have been established by the Food and Nutrition Board, Institute of Medicine, at 1000 mg for vitamin E and 2000 mg for vitamin C in adults. Many clinical trials with these vitamins have involved subjects with various diseases, and no consistent pattern of adverse effects has occurred at any intake. Numerous studies of vitamin C supplementation have provided no pattern of evidence to support concerns about safety other than occasional gastrointestinal upset or mild diarrhea resulting from the osmotic effects of unabsorbed quantities of vitamin C. Evidence of bleeding effects and other potential adverse effects of high vitamin E intakes in humans is not convincing. Evidence of adverse effects of vitamin C that result from its effects on iron absorption and metabolism has not been confirmed in clinical trials. Thus, we conclude from clinical trial evidence that vitamin E supplements appear safe for most adults in amounts 88 cm in women and >102 cm in men) and 260 with peripheral fat distribution (PFD). Fat-free mass (FFM) and fat mass (FM) were measured by bioelectric impedance. Body composition was estimated as FM/FFM. Echocardiographic stroke volume (SV) and cardiac output (CO) were measured. RESULTS: Hypertension was present in 73% of the subjects with PFD and in 78% with CFD. Overweight with CFD was associated with greater FM/FFM in both normotensive and hypertensive participants. After FFM, age, sex, and race were controlled for, SV and CO were higher in subjects overweight with CFD than in those with PFD, whereas peripheral resistance was not significantly different. Differences in CO between CFD and PFD were reduced after further adjustment for FM. After the covariates were controlled for, hypertensive subjects had higher peripheral resistance and lower arterial compliance than did normotensive participants, but cardiac output was not significantly different. CONCLUSION: CFD is associated with more severe abnormalities in body composition and with higher CO independently of FFM in overweight, nonobese subjects. PMID- 15817850 TI - Carbohydrate-restricted diets high in either monounsaturated fat or protein are equally effective at promoting fat loss and improving blood lipids. AB - BACKGROUND: When substituted for carbohydrate in an energy-reduced diet, dietary protein enhances fat loss in women. It is unknown whether the effect is due to increased protein or reduced carbohydrate. OBJECTIVE: We compared the effects of 2 isocaloric diets that differed in protein and fat content on weight loss, lipids, appetite regulation, and energy expenditure after test meals. DESIGN: This was a parallel, randomized study in which subjects received either a low fat, high-protein (LF-HP) diet (29 +/- 1% fat, 34 +/- 0.8% protein) or a high fat, standard-protein (HF-SP) diet (45 +/- 0.6% fat, 18 +/- 0.3% protein) during 12 wk of energy restriction (6 +/- 0.1 MJ/d) and 4 wk of energy balance (7.4 +/- 0.3 MJ/d). Fifty-seven overweight and obese [mean body mass index (in kg/m(2)): 33.8 +/- 0.9] volunteers with insulin concentrations >12 mU/L completed the study. RESULTS: Weight loss (LF-HP group, 9.7 +/- 1.1 kg; HF-SP group, 10.2 +/- 1.4 kg; P = 0.78) and fat loss were not significantly different between diet groups even though the subjects desired less to eat after the LF-HP meal (P = 0.02). The decrease in resting energy expenditure was not significantly different between diet groups (LF-HP, -342 +/- 185 kJ/d; HF-SP, -349 +/- 220 kJ/d). The decrease in the thermic effect of feeding with weight loss was smaller in the LF HP group than in the HF-SP group (-0.3 +/- 1.0% compared with -3.6 +/- 0.7%; P = 0.014). Glucose and insulin responses to test meals improved after weight loss (P < 0.001) with no significant diet effect. Bone turnover, inflammation, and calcium excretion did not change significantly. CONCLUSION: The magnitude of weight loss and the improvements in insulin resistance and cardiovascular disease risk factors did not differ significantly between the 2 diets, and neither diet had any detrimental effects on bone turnover or renal function. PMID- 15817851 TI - Dietary fat intake and early age-related lens opacities. AB - BACKGROUND: Dietary fat may affect lens cell membrane composition and function, which are related to age-related cataract. OBJECTIVE: The objective of the study was to examine the association between long-term dietary fat intake and the prevalence of age-related nuclear, cortical, and posterior subcapsular lens opacities. DESIGN: Women (n = 440) aged 53-73 y from the Boston area without previously diagnosed cancer, diabetes, or cataract were selected from the Nurses' Health Study cohort. Intakes of total fat and selected fatty acids were calculated as the average of intake data from 5 food-frequency questionnaires collected between 1980 and the study eye examination (1993-1995). Nuclear opacity was defined as grade >/=2.5, cortical opacity as grade >/=1.0, and posterior subcapsular opacity as grade >/=0.5 according to the Lens Opacities Classification System III. RESULTS: There were significant positive associations between linoleic and linolenic acid intakes and the prevalence of nuclear opacity. The odds ratios for nuclear opacity in women with intakes in the highest quartile and women with intakes in the lowest quartile were 2.2 (95% CI: 1.1, 4.6; P for trend = 0.02) for linoleic acid and 2.2 (95% CI: 1.1, 4.5; P for trend = 0.05) for linolenic acid. There were no significant associations between intakes of any type of fat and either cortical or posterior subscapular opacity. CONCLUSIONS: High intake of the 18-carbon polyunsaturated fatty acids linoleic acid and linolenic acid may increase the risk of age-related nuclear opacity. Further study is needed to clarify the relation between dietary fat and cataract risk. PMID- 15817852 TI - Association between dietary factors and plasma adiponectin concentrations in men. AB - BACKGROUND: Adiponectin, an adipocyte-derived peptide, improves insulin sensitivity, has antiinflammatory and antiatherogenic effects, and is associated with a lower risk of ischemic heart disease (IHD) and type 2 diabetes. However, little is known about dietary predictors of plasma adiponectin concentrations in humans. OBJECTIVE: Our objective was to examine cross-sectionally the association between dietary factors and plasma adiponectin in men. DESIGN: Our study included 532 male participants of the Health Professionals Follow-Up Study who were selected as control subjects for an investigation of biological predictors of IHD. Diet, lifestyle, and anthropometric data were assessed by questionnaires. RESULTS: After multivariable adjustment, adiponectin was significantly inversely related to glycemic load (-1.3 mg/L per 1-SD increase; P = 0.02) and tended to be positively associated with total fat intake (0.7 mg/L per 0.5% of energy from fat instead of carbohydrates; P = 0.06). We also found a significant nonlinear association between plasma adiponectin concentrations and alcohol intake (P for quadratic trend = 0.01). Thus, whereas nondrinkers had mean plasma adiponectin concentrations of 16.48 mg/L, those who consumed 0.1-4.9, 5.0-14.9, 15.0-29.9, or >/=30 g alcohol/d had mean concentrations of 16.79 (P = 0.77 compared with nondrinkers), 18.97 (P = 0.02), 19.11 (P = 0.01), and 18.39 (P = 0.10) mg/L, respectively. CONCLUSIONS: Moderate alcohol intake is associated with higher adiponectin concentrations, whereas a carbohydrate-rich diet with a high glycemic load is associated with lower adiponectin concentrations in men with no history of cardiovascular disease. Although the strength of these associations was modest, our observations highlight the hypothesis that dietary factors may modulate plasma adiponectin concentrations-a potential mediator related to a reduced IHD risk. PMID- 15817853 TI - Iron, zinc, and alcohol consumption and mortality from cardiovascular diseases: the Iowa Women's Health Study. AB - BACKGROUND: The relation between iron status and atherosclerosis has long been a topic of debate. OBJECTIVE: We examined associations of cardiovascular disease (CVD) mortality with dietary intakes of iron (a possible prooxidant), zinc (a possible antioxidant), and alcohol (a disruptor of iron homeostasis). DESIGN: Postmenopausal women (n = 34 492) aged 55-69 y at baseline, who completed a food frequency questionnaire, were followed for CVD mortality over 15 y. RESULTS: Among women who consumed >/=10 g alcohol/d, after adjustment for CVD risk factors in a model that contained dietary heme iron, nonheme iron, and zinc intakes, dietary heme iron showed a positive association, dietary nonheme iron showed a U shaped association, and dietary zinc showed an inverse association with CVD mortality. For example, the relative risks (RRs) for categories of dietary heme iron were 1.0, 1.46, 1.52, 1.73, and 2.47 (P for trend = 0.04); corresponding RRs for dietary nonheme iron were 1.0, 0.93, 0.63, 0.83, and 1.20 (P for quadratic term = 0.02). The corresponding RRs for dietary zinc were 1.0, 0.61, 0.59, 0.57, and 0.37 (P for trend = 0.07). In an analysis restricted to those who consumed >/=30 g alcohol/d, the risk gradients strengthened. CONCLUSIONS: Our results suggest that a higher intake of heme iron might be harmful, whereas a higher intake of zinc might be beneficial in relation to CVD mortality in the presence of a trigger that can disturb iron homeostasis, such as alcohol consumption. PMID- 15817854 TI - Cholesterol-lowering effects of bovine serum immunoglobulin in participants with mild hypercholesterolemia. AB - BACKGROUND: The consumption of milk products has been shown to lower cholesterol. The mechanism of action surrounding this observation has been attributed to the protein fraction of milk and serum. OBJECTIVE: We examined the effect of an oral serum bovine immunoglobulin protein fraction (bIg) derived from US Department of Agriculture-approved beef (aged <30 mo) on lipid indexes in humans. DESIGN: Participants included men and women (aged 25-70 y) with hypercholesterolemia (5.44-6.99 mmol/L) who were not receiving cholesterol-lowering medication. Treatment consisted of the randomized, double-blind, parallel-group, placebo controlled administration of 5 g bIg for 6 wk in 52 participants (n = 26 each in treatment and control groups). RESULTS: Mean (+/-SD) total cholesterol (TC) at baseline was 6.33 +/- 0.1 mmol/L for bIg and 6.16 +/- 0.1 mmol/L for placebo. A repeated-measures multivariate analysis of covariance covaried for change in total energy and alcohol intake and Tukey's post hoc examination of our data showed that the bIg-treated group had a significant reduction in TC at 3 wk (5.98 +/- 0.5 mmol/L; P < 0.05) and 6 wk (5.97 +/- 0.7 mmol/L; P < 0.05). The concentration at 6 wk was significantly different from that in the placebo group (P < 0.05). This reduction was largely due to a decrease in LDL cholesterol in the bIg-treated group from baseline (4.12 +/- 0.6 mmol/L) at 3 wk (3.92 +/- 0.7 mmol/L, P < 0.05) and at 6 wk (3.84 +/- 0.6 mmol/L, P < 0.05); the 6-wk concentration differed significantly between the treatment and placebo groups (P < 0.05). We observed no significant changes in the placebo group or in any other lipid indexes or markers associated with hepatorenal or cardiovascular function. CONCLUSION: Consumption of bIg appears to positively modulate the primary lipid indexes associated with cardiovascular disease. PMID- 15817855 TI - A cellular-level approach to predicting resting energy expenditure across the adult years. AB - BACKGROUND: We previously derived a whole-body resting energy expenditure (REE) prediction model by using organ and tissue mass measured by magnetic resonance imaging combined with assumed stable, specific resting metabolic rates of individual organs and tissues. Although the model predicted REE well in young persons, it overpredicted REE by approximately 11% in elderly adults. This overprediction may occur because of a decline in the fraction of organs and tissues as cell mass with aging. OBJECTIVE: The aim of the present study was to develop a cellular-level REE prediction model that would be applicable across the adult age span. Specifically, we tested the hypothesis that REE can be predicted from a combination of organ and tissue mass, the specific resting metabolic rates of individual organs and tissues, and the cellular fraction of fat-free mass. DESIGN: Fifty-four healthy subjects aged 23-88 y had REE, organ and tissue mass, body cell mass, and fat-free mass measured by indirect calorimetry, magnetic resonance imaging, whole-body (40)K counting, and dual-energy X-ray absoptiometry, respectively. RESULTS: REE predicted by the cellular-level model was highly correlated with measured REE (r = 0.92, P < 0.001). The mean difference between measured REE (x+/- SD: 1487 +/- 294 kcal/d) and predicted REE (1501 +/- 300 kcal/d) for the whole group was not significant, and the difference between predicted and measured REE was not associated with age (r = 0.009, NS). CONCLUSION: The present approach establishes an REE-body composition link with the use of a model at the cellular level. The combination of 2 aging-related factors (ie, decline in both the mass and the cellular fraction of organs and tissues) may account for the lower REE observed in elderly adults. PMID- 15817856 TI - Use of mathematical modeling to study copper metabolism in humans. AB - BACKGROUND: An improved understanding of copper metabolism is needed to derive more precise estimates of dietary requirements. OBJECTIVES: The objectives were to validate a method for estimating endogenous losses of copper, test whether a simple model can predict true absorption from the plasma appearance of labeled copper, and develop a compartmental model for copper metabolism by using stable isotopes. DESIGN: A stable isotope of copper was intravenously administered to 6 men, and fecal samples were collected for 14 d. Four weeks later the study was repeated, but with an oral dose, and blood samples were collected for 7 d and fecal samples for 14 d. RESULTS: There was no significant difference (P = 0.48) in the estimated endogenous loss of copper calculated by using either the excreted intravenous dose (x +/- SD: 32 +/- 5%) or the absorbed and excreted oral dose (35 +/- 2%). A simple mathematical model fitted to plasma isotope appearance data estimated true absorption to be 8 +/- 2% compared with 48-49% measured by fecal monitoring. A more complicated compartmental model predicted that, when newly absorbed copper first enters the blood, 74% is removed by the liver and 99% is bound to ceruloplasmin in the plasma. The exchangeable pool of copper was estimated to be 43 +/- 30 mg. Daily endogenous losses were predicted to be 2.4 mg. CONCLUSIONS: The results showed that fecal monitoring is the only method that can reliably measure labeled copper absorption, and it is not necessary to administer an intravenous dose of copper to estimate endogenous losses. The compartmental model provides new insights into human copper metabolism. PMID- 15817857 TI - Iron absorption in male C282Y heterozygotes. AB - BACKGROUND: The suggestion that carriers of the HFE C282Y mutation absorb nonheme iron more efficiently than do carriers of the wild type has public health implications for countries where the C282Y mutation is common and foods are fortified with iron. OBJECTIVE: We investigated the effect of C282Y heterozygosity on nonheme-iron absorption from a diet high in bioavailable iron and from iron-fortified cereals. DESIGN: The subjects were recruited from a parallel study investigating the relation between HFE mutations, habitual diet, and iron status. Iron absorption was measured in 15 wild-type carriers and 15 C282Y heterozygotes aged >/=40 y. Each subject consumed 3 meals of high iron bioavailability (labeled with Fe-57) for 2 d and 2 meals with fortified cereal products (labeled with Fe-54) for the next 3 d. Iron absorption was measured from isotope incorporation into red blood cells 14 d after the last labeled meal and was corrected for utilization of absorbed iron by means of an intravenous infusion of Fe-58. RESULTS: Absorption of Fe-57 with the high-iron bioavailability diet was 6.8 +/- 6.8% (0.6 +/- 0.6 mg/d) in the wild-type carriers and 7.6 +/- 3.2% (0.7 +/- 0.3 mg/d) in the C282Y heterozygotes. Absorption of Fe-54 with cereal products was 4.9 +/- 2.0% (0.7 +/- 0.3 mg/d) in the wild-type carriers and 5.3 +/- 1.3% (0.8 +/- 0.2 mg/d) in the C282Y heterozygotes. CONCLUSIONS: There was no overall significant difference between C282Y heterozygotes and wild-type men in iron absorption from either dietary nonheme iron or fortified cereal products. PMID- 15817858 TI - Long-term high copper intake: effects on copper absorption, retention, and homeostasis in men. AB - BACKGROUND: Numerous studies have examined the effect of low and adequate intakes of copper on absorption and retention, but little information is available on the regulation of absorption and retention of copper when intake is high. OBJECTIVE: A study was conducted in men to determine the effect of long-term high copper intake on copper absorption, retention, and homeostasis. DESIGN: Nine men were confined to a metabolic research unit (MRU) for 18 d and were fed a 3-d rotating menu containing an average of 1.6 mg Cu/d. They continued the study under free living conditions for 129 d, supplementing their usual diets with 7 mg Cu/d. They then returned to the MRU for 18 d and consumed the same diet as during the first period, except that copper intake was 7.8 mg/d. The stable isotope (63)Cu was fed to 3 subjects and infused into the other 6 on day 7 of each MRU period, and complete urine and stool collections were made throughout the study. Total copper and (63)Cu were determined by inductively coupled plasma mass spectrometry. Copper absorption, excretion, and retention were calculated on the basis of dietary, urinary, and fecal copper and (63)Cu. RESULTS: Results were as follows when comparing the high copper intake with the usual intake: fractional copper absorption was significantly lower, but the amount absorbed was significantly higher; excretion of the infused (63)Cu was significantly faster; and total retention was significantly higher. CONCLUSIONS: Homeostatic regulation of copper absorption and retention helped to minimize the amount of copper retained with high copper intake but was not sufficient to prevent retention of >0.6 mg Cu/d. PMID- 15817859 TI - Effectiveness of selenium supplements in a low-selenium area of China. AB - BACKGROUND: Selenium is an essential micronutrient with a recommended dietary allowance for adults of 55 mug/d. It functions as an essential constituent of selenoproteins. Although there is no evidence of selenium deficiency in the United States, people in many other areas of the world are selenium deficient, with the consequence that they are unable to express their selenoproteins fully. OBJECTIVE: We carried out a supplementation trial in a selenium-deficient population in China to assess the requirement for selenium as selenite and as selenomethionine. DESIGN: One hundred twenty subjects with an average selenium intake of 10 mug/d were randomly assigned and administered tablets containing no selenium or amounts as high as 66 mug Se/d for 20 wk. Plasma was sampled before supplementation and at 4-wk intervals during supplementation and was assayed for the 2 plasma selenoproteins, glutathione peroxidase and selenoprotein P. RESULTS: Full expression of glutathione peroxidase was achieved with 37 mug Se/d as selenomethionine and with 66 mug/d as selenite. Full expression of selenoprotein P was not achieved at the highest doses of either form. CONCLUSIONS: Full expression of selenoprotein P requires a greater selenium intake than does full expression of plasma glutathione peroxidase. This suggests that selenoprotein P is a better indicator of selenium nutritional status than is glutathione peroxidase and that the recommended dietary allowance of selenium, which was set with the use of glutathione peroxidase as the index of selenium status, should be revised. Selenium as selenomethionine had nearly twice the bioavailability of selenium as selenite. PMID- 15817860 TI - Validity of the relative-dose-response test and the modified-relative-dose response test as indicators of vitamin A stores in liver. AB - BACKGROUND: Our group and many others have used the relative-dose-response (RDR) test and the modified-RDR (MRDR) test as proxy indicators of liver stores of vitamin A. However, we have become concerned about the validity of these indicators. OBJECTIVE: Simulation models were used to assess effects of random variations in serum retinol concentration on the RDR and to assess effects of group differences in serum retinol concentration on the distribution of RDR and MRDR values. DESIGN: Random and independent samples were drawn from normally distributed, computer-generated numbers whose distributions simulated serum concentrations of retinol and 3,4-didehydroretinol as obtained from published reports. The resulting data sets were used to compute surrogate RDR or MRDR values. In model 1, the relation between serum concentrations of retinol and RDR was examined within a fictitious population. In models 2 and 3, fictitious populations with different distributions of serum retinol concentration were compared with respect to their RDR and MRDR values. RESULTS: Simulated RDR values and serum retinol concentrations were negatively related. Models 2 and 3 showed that group differences in serum retinol concentrations necessarily produced group differences in mean RDR or MRDR values. A mathematical artifact may explain the negative relation reported between MRDR and serum retinol concentration, and it dictates that this relation will necessarily vary between populations with different degrees of vitamin A deficiency. CONCLUSION: A continued search for alternative blood indicators of liver stores of vitamin A is needed. PMID- 15817861 TI - High thyroid volume in children with excess dietary iodine intakes. AB - BACKGROUND: There are few data on the adverse effects of chronic exposure to high iodine intakes, particularly in children. OBJECTIVE: The objective of the study was to ascertain whether high dietary intakes of iodine in children result in high thyroid volume (Tvol), a high risk of goiter, or both. DESIGN: In an international sample of 6-12-y-old children (n = 3319) from 5 continents with iodine intakes ranging from adequate to excessive, Tvol was measured by ultrasound, and the urinary iodine (UI) concentration was measured. Regressions were done on Tvol and goiter including age, body surface area, sex, and UI concentration as covariates. RESULTS: The median UI concentration ranged from 115 microg/L in central Switzerland to 728 microg/L in coastal Hokkaido, Japan. In the entire sample, 31% of children had UI concentrations >300 microg/L, and 11% had UI concentrations >500 microg/L; in coastal Hokkaido, 59% had UI concentrations >500 microg/L, and 39% had UI concentrations >1000 microg/L. In coastal Hokkaido, the mean age- and body surface area-adjusted Tvol was approximately 2-fold the mean Tvol from the other sites combined (P < 0.0001), and there was a positive correlation between log(UI concentration) and log(Tvol) (r = 0.24, P < 0.0001). In the combined sample, after adjustment for age, sex, and body surface area, log(Tvol) began to rise at a log(UI concentration) >2.7, which, when transformed back to the linear scale, corresponded to a UI concentration of approximately 500 microg/L. CONCLUSIONS: Chronic iodine intakes approximately twice those recommended-indicated by UI concentrations in the range of 300-500 microg/L-do not increase Tvol in children. However, UI concentrations >/=500 microg/L are associated with increasing Tvol, which reflects the adverse effects of chronic iodine excess. PMID- 15817862 TI - Racial differences in calcium retention in response to dietary salt in adolescent girls. AB - BACKGROUND: Sodium is an important determinant of urinary calcium excretion, and race is an important determinant of calcium retention. The effect of dietary sodium on calcium retention and the influence of race have not been studied in adolescence, the life stage during which peak bone mass is accrued. OBJECTIVE: The study reported here was undertaken to compare racial differences in calcium retention as a function of dietary salt intake. DESIGN: A total of 35 adolescent girls (22 black and 13 white) participated in two 20-d metabolic summer camps, separated by 2 wk, that simulated a free-living environment. The effect of changes in dietary sodium on calcium retention was tested in a randomized-order, crossover design with 2 concentrations of sodium-1.30 g/d (57 mmol/d) and 3.86 g/d (168 mmol/d)-and a constant calcium intake of 815 mg/d (20 mmol/d). RESULTS: Both race and sodium intake significantly affected calcium retention (P < 0.01). Calcium retention was significantly greater in black girls than in white girls, regardless of dietary sodium intake (P < 0.001). The high-sodium diet significantly reduced calcium retention in both whites and blacks (P < 0.01), primarily through a decrease in net calcium absorption. Black girls excreted significantly less calcium in the urine than did white girls, regardless of diet (P < 0.05). CONCLUSIONS: Calcium retention is significantly greater in black girls than in white girls but is significantly reduced in girls of both races in response to salt loading. PMID- 15817863 TI - Elevated serum concentrations of beta-glucuronide metabolites and 4-oxoretinol in lactating sows after treatment with vitamin A: a model for evaluating supplementation in lactating women. AB - BACKGROUND: The effects of large doses of preformed vitamin A, such as those provided in supplementation programs for the prevention of deficiency, on total serum vitamin A have been inadequately investigated. OBJECTIVE: This study characterized the time course of serum vitamin A metabolites in lactating sows after single high doses of retinyl ester. DESIGN: Lactating sows were fitted with jugular catheters and subsequently fed either 1.05 or 2.1 mmol retinyl ester (n = 6/group) or a corn oil vehicle (n = 3). Blood was collected at baseline and at intervals to 48 h and analyzed by gradient HPLC for retinol, retinyl esters, and metabolites. RESULTS: The mean (+/-SD) total serum vitamin A concentration peaked at 1 h (3.69 +/- 4.0 mumol/L) and 2 h (7.70 +/- 6.8 mumol/L) in the low- and high dose groups, respectively (P < 0.05). Retinyl esters accounted for most of the serum vitamin A in both groups at peak time points. Mean serum retinol concentrations changed little and accounted for most of the serum vitamin A at baseline (94% and 97% for the low- and high-dose groups, respectively) but for only 22% and 14% at peak times for the low- and high-dose groups, respectively. Postdosage increases were observed for total vitamin A and retinyl esters, 4 oxoretinol, retinoyl beta-glucuronide, and retinyl beta-glucuronide but not for retinoic acid. CONCLUSIONS: Serum retinol concentration remains relatively static after a large dose of preformed vitamin A and therefore is not an appropriate measure of intervention efficacy. The increases in beta-glucuronide metabolites and 4-oxoretinol suggest a preventive role against a rise in retinoic acid and retinol. PMID- 15817864 TI - Vitamin C supplementation to prevent premature rupture of the chorioamniotic membranes: a randomized trial. AB - BACKGROUND: Vitamin C is involved in the synthesis and degradation of collagen and is important for maintenance of the chorioamniotic membranes. Inadequate availability of ascorbic acid during pregnancy has been proposed as a risk factor for premature rupture of the chorioamniotic membranes (PROM). OBJECTIVE: The objective of the study was to evaluate the effectiveness of 100 mg vitamin C/d in preventing PROM. DESIGN: A controlled double-blind trial was performed. Pregnant women (n = 126) in their 20th wk of gestation were invited; 120 accepted and were randomly assigned to 2 groups (100 mg vitamin C/d or placebo). Every 4 wk, plasma and leukocyte vitamin C concentrations were measured, and each subject was evaluated for cervicovaginal infection. The incidence of PROM was recorded for each group as an indicator of the protective effect of vitamin C supplementation. RESULTS: One hundred nine patients finished the study. Mean plasma vitamin C concentrations decreased significantly throughout the pregnancy in both groups (P = 0.001), and there were no significant differences between groups. Between weeks 20 and 36, mean leukocyte vitamin C concentrations decreased from 17.5 to 15.23 microg/10(8) cells in the placebo group and increased from 17.26 to 22.17 microg/10(8) cells in the supplemented group (within- and between-group differences: P = 0.001). The incidence of PROM was 14 per 57 pregnancies (24.5%) in the placebo group and 4 per 52 pregnancies (7.69%) in the supplemented group (relative risk: 0.26; 95% CI: 0.078, 0.837). CONCLUSION: Daily supplementation with 100 mg vitamin C after 20 wk of gestation effectively lessens the incidence of PROM. PMID- 15817865 TI - Comparison of home-based therapy with ready-to-use therapeutic food with standard therapy in the treatment of malnourished Malawian children: a controlled, clinical effectiveness trial. AB - BACKGROUND: Childhood malnutrition is common in Malawi, and the standard treatment, which follows international guidelines, results in poor recovery rates. Higher recovery rates have been seen in pilot studies of home-based therapy with ready-to-use therapeutic food (RUTF). OBJECTIVE: The objective was to compare the recovery rates among children with moderate and severe wasting, kwashiorkor, or both receiving either home-based therapy with RUTF or standard inpatient therapy. DESIGN: A controlled, comparative, clinical effectiveness trial was conducted in southern Malawi with 1178 malnourished children. Children were systematically allocated to either standard therapy (186 children) or home based therapy with RUTF (992 children) according to a stepped wedge design to control for bias introduced by the season of the year. Recovery, defined as reaching a weight-for-height z score > -2, and relapse or death were the primary outcomes. The rate of weight gain and the prevalence of fever, cough, and diarrhea were the secondary outcomes. RESULTS: Children who received home-based therapy with RUTF were more likely to achieve a weight-for-height z score > -2 than were those who received standard therapy (79% compared with 46%; P < 0.001) and were less likely to relapse or die (8.7% compared with 16.7%; P < 0.001). Children who received home-based therapy with RUTF had greater rates of weight gain (3.5 compared with 2.0 g . kg(-1) . d(-1); difference: 1.5; 95% CI: 1.0, 2.0 g . kg(-1) . d(-1)) and a lower prevalence of fever, cough, and diarrhea than did children who received standard therapy. CONCLUSION: Home-based therapy with RUTF is associated with better outcomes for childhood malnutrition than is standard therapy. PMID- 15817866 TI - Visual maturation of term infants fed long-chain polyunsaturated fatty acid supplemented or control formula for 12 mo. AB - BACKGROUND: Several studies found a benefit of long-chain polyunsaturated fatty acid (LCP) supplementation for visual or mental development, but others found no benefit. Likely contributors to differences among studies are the amount of LCP supplementation, functional outcomes, and sample size. OBJECTIVE: We evaluated LCP supplementation in amounts typical for human milk (based on local and worldwide surveys) in a large cohort of infants by using sweep visual evoked potential (VEP) acuity as the functional outcome. DESIGN: The study was a double masked, randomized, controlled clinical trial in 103 term infants. By age 5 d, infants were randomly assigned to receive either formula with no docosahexaenoic acid (DHA) or arachidonic acid (ARA) or formula supplemented with DHA and ARA as 0.36% and 0.72%, respectively, of total fatty acids. Sweep VEP acuity was the primary outcome. Random dot stereoacuity, blood lipid profile, growth, and tolerance were secondary outcomes. RESULTS: VEP acuity in the LCP-supplemented group was significantly better than that in the control group at ages 6, 17, 26, and 52 wk. Stereoacuity in the LCP-supplemented group was significantly better than that in the control group at age 17 wk but not at ages 39 and 52 wk. By ages 17 and 39 wk, the red blood cell DHA concentration in the LCP-supplemented group was more than double and more than triple, respectively, that in the control group. Growth of infants fed LCP-supplemented and control formulas did not differ significantly, and both diets were well tolerated. CONCLUSION: LCP supplementation of term infant formula during the first year of life yields clear differences in visual function and in total red blood cell lipid composition. PMID- 15817867 TI - Vitamin supplementation of HIV-infected women improves postnatal child growth. AB - BACKGROUND: Linear growth retardation and wasting are common in children born to HIV-infected women. Inexpensive interventions that could improve the postnatal growth pattern of such children are needed. OBJECTIVE: The objective was to examine the effect of supplementing HIV-infected women with multivitamins or vitamin A and beta-carotene, during and after pregnancy, on the growth of their children during the first 2 y of life. DESIGN: We conducted a randomized placebo controlled trial in 886 mother-infant pairs in Tanzania. At the first prenatal visit, HIV-infected women were randomly assigned to 1 of 4 daily oral regimens in a 2 x 2 factorial fashion: multivitamins (MV: thiamine, riboflavin, vitamin B-6, niacin, vitamin B-12, vitamin C, vitamin E, and folic acid), preformed vitamin A + beta-carotene (VA/BC), MV including VA/BC, or placebo. Supplementation continued during the first 2 y postpartum and thereafter. Children were weighed and measured monthly, and all received vitamin A supplements after 6 mo of age per the standard of care. RESULTS: Multivitamins had a significant positive effect on attained weight (459 g; 95% CI: 35, 882; P = 0.03) and on weight-for age (0.42; 95% CI: 0.07, 0.77; P = 0.02) and weight-for-length (0.38; 95% CI: 0.07, 0.68; P = 0.01) z scores at 24 mo. VA/BC seemed to reduce the benefits of MV on these outcomes. No significant effects were observed on length, midupper arm circumference, or head circumference. CONCLUSION: Supplementation of HIV infected women with multivitamins (vitamin B complex, vitamin C, and vitamin E) during pregnancy and lactation is an effective intervention for improving ponderal growth in children. PMID- 15817868 TI - Leptin and body weight regulation in patients with anorexia nervosa before and during weight recovery. AB - BACKGROUND: Leptin has been considered a starvation hormone, but its role in malnourished patients is unknown. OBJECTIVE: We aimed to characterize the role of leptin in metabolic adaptation in women with anorexia nervosa (AN). DESIGN: In a cross-sectional study, 57 women with AN [mean (+/-SD) body mass index (kg/m(2)) on admission: 15.2 +/- 1.5] were compared with 49 healthy, normal-weight women (mean body mass index: 22.3 +/- 2.3). Nineteen patients were reinvestigated during weight gain 43 and 84 d after baseline. We measured serum concentrations of leptin, soluble leptin receptor, insulin, ghrelin, and thyroid hormones [thyrotropin, triiodothyronine (T(3)), and thyroxine]; fat mass (FM) and fat-free mass (FFM); resting energy expenditure (REE); energy intake; and eating behavior. RESULTS: Compared with values in the control women, leptin, T(3), REE, FM, and FFM were lower in the women with AN, but the leptin secretion rate was not significantly different. Leptin correlated with FM (r = 0.83, P < 0.001), T(3) (r = 0.68, P < 0.001), respiratory quotient (r = -0.47, P < 0.001), and REE (r = 0.58, P < 0.001). The association with REE weakened after adjustment for FFM and disappeared after further adjustment for T(3). Hunger and appetite had positive, whereas satiety and restraint had negative, associations with leptin. During weight gain (9.0 +/- 3.3 kg in 84 d), serum leptin and the leptin secretion rate increased. Changes in leptin secretion were associated with energy intake and REE. The initial changes in the leptin secretion rate (ie, the difference between baseline and 43 d) were negatively associated with changes in body weight from 43 to 84 d. CONCLUSIONS: Leptin contributes to metabolic adaptation in women with AN. The leptin response is associated with weight gain. PMID- 15817869 TI - Cognitive impairment and composition of drinking water in women: findings of the EPIDOS Study. AB - BACKGROUND: The concentration of aluminum or silica in drinking water may be a potential environmental risk factor for Alzheimer disease (AD). OBJECTIVES: The objective was to investigate at baseline the potential association between the composition of drinking water and the level of cognitive function in women taking part in the Epidemiology of Osteoporosis (EPIDOS) Study and to determine during follow-up the effects of the composition of drinking water on the risk of AD. DESIGN: Women aged >/=75 y (n = 7598) were recruited between 1992 and 1994 in 5 geographic areas of France. The participants from one center (n = 1462) were followed for T polymorphism in determining hepatic lipase activity: the Hoorn Study. AB - BACKGROUND: Gene-nutrient interactions affecting hepatic lipase (HL) activity may contribute to the interindividual variability of the cardiovascular disease risk associated with dietary fat intake. OBJECTIVE: We determined the associations of dietary fat intake with postheparin HL activity and the possible modifying effect of the HL -480C-->T polymorphism on these associations. DESIGN: Subjects were recruited from participants in the 2000-2001 follow-up examination of the Hoorn Study. HL activity was determined in postheparin plasma in a sample of 211 men and 218 women aged 60-87 y. Information about dietary intake of the participants was obtained with a validated food-frequency questionnaire. Linear regression was performed, adjusted for age. RESULTS: Total dietary fat was positively associated with HL activity (standardized beta: 0.11; 95% CI: 0.02, 0.21), and this association was also seen for saturated fat (0.10; 0.01, 0.20) and monounsaturated fatty acid (0.10; 0.01, 0.19). We observed a significant interaction of the HL polymorphism with the relation between total fat intake and HL activity. The association of total fat with HL activity was stronger in subjects with CT (0.27; 0.11, 0.43) and TT (0.39; -0.22, 1.00) genotypes than in subjects with the CC genotype (0.06; -0.06, 0.18; P for interaction < 0.10). The interaction remained statistically significant in models that included age, sex, carbohydrate and protein intakes, and insulin or body mass index. CONCLUSIONS: Higher intakes of total and saturated fat were positively associated with higher HL activity. In addition, the observed association of total fat with HL activity was modified by the HL-480C-->T polymorphism, after adjustment for age, sex, carbohydrate and protein intakes, and insulin or body mass index. PMID- 15817872 TI - The effect of soy protein and soy isoflavones on calcium metabolism in postmenopausal women: a randomized crossover study. AB - BACKGROUND: Evidence suggests that soy isoflavones act as estrogen agonists and have beneficial skeletal effects, but the effects on calcium metabolism in humans are not known. OBJECTIVE: This study tested whether soybean isoflavones, soy protein, or both alter calcium metabolism in postmenopausal women. DESIGN: Calcium metabolism in 15 postmenopausal women was studied by using metabolic balance and kinetic modeling in a randomized, crossover design of three 1-mo controlled dietary interventions: soy protein isolate enriched with isoflavones (soy-plus diet), soy protein isolate devoid of isoflavones (soy-minus diet), and a casein-whey protein isolate (control diet). RESULTS: There was no significant difference between the diets in net acid excretion (P = 0.12). Urinary calcium excretion was significantly (P < 0.01) less with consumption of either of the soy diets (soy-plus diet: 85 +/- 34 mg/d; soy-minus diet: 80 +/- 34 mg/d) than with consumption of the control diet (121 +/- 63 mg/d), but fractional calcium absorption was unaffected by treatment. Endogenous fecal calcium was significantly (P < 0.01) greater with consumption of the soy-minus diet than with consumption of the other diets. Total fecal calcium excretion, bone deposition and resorption, and calcium retention were not significantly affected by the dietary regimens. CONCLUSIONS: The lower urinary calcium seen with the consumption of an isolated soy protein than with that of an isolated milk protein was not associated with improved calcium retention. This finding reinforces the importance of evaluating all aspects of calcium metabolism. Soy isoflavones did not significantly affect calcium metabolism. PMID- 15817873 TI - Low dietary potassium intakes and high dietary estimates of net endogenous acid production are associated with low bone mineral density in premenopausal women and increased markers of bone resorption in postmenopausal women. AB - BACKGROUND: The Western diet may be a risk factor for osteoporosis. Excess acid generated from high protein intakes increases calcium excretion and bone resorption. Fruit and vegetable intake could balance this excess acidity by providing alkaline salts of potassium. Algorithms based on dietary intakes of key nutrients can be used to approximate net endogenous acid production (NEAP) and to explore the association between dietary acidity and bone health. OBJECTIVE: We investigated the relation between dietary potassium and protein, NEAP (with an algorithm including the ratio of protein to potassium intake), and potential renal acid load (with an algorithm including dietary protein, phosphorous, potassium, magnesium, and calcium) and markers of bone health. DESIGN: Measurements of bone mineral density (BMD) (n = 3226) and urinary bone resorption markers (n = 2929) at the lumbar spine and femoral neck were performed in perimenopausal and early postmenopausal women aged 54.9 +/- 2.2 y (x +/- SD) in 1997-1999. BMD (g/cm(2)), free pyridinoline (fPYD), and free deoxypyridinoline (fDPD) were expressed relative to creatinine. Dietary intake was assessed with a food-frequency questionnaire. RESULTS: Comparison of the highest with the lowest quartile of potassium intake or the lowest with the highest NEAP showed a 6-8% increase in fPYD/creatinine and fDPD/creatinine. A difference of 8% in BMD was observed between the highest and lowest quartiles of potassium intake in the premenopausal group (n = 337). CONCLUSIONS: Dietary potassium, an indicator of NEAP and fruit and vegetable intake, may exert a modest influence on markers of bone health, which over a lifetime may contribute to a decreased risk of osteoporosis. PMID- 15817874 TI - Ratio of n-6 to n-3 fatty acids and bone mineral density in older adults: the Rancho Bernardo Study. AB - BACKGROUND: Several lines of evidence suggest that n-3 fatty acids reduce the risk of some chronic diseases, including heart disease, diabetes, and cancer. Other research, mainly in animals, also suggests a role in bone health. OBJECTIVE: We aimed to investigate the association between the ratio of dietary n 6 to n-3 fatty acids and bone mineral density (BMD) in 1532 community-dwelling men and women aged 45-90 y. DESIGN: Between 1988 and 1992, dietary data were obtained through self-administered food-frequency questionnaires, and BMD was measured at the hip and spine with the use of dual-energy X-ray absorptiometry. A medical history was obtained and current medication use was validated. Age- and multiple-adjusted linear regression analyses were performed. RESULTS: There was a significant inverse association between the ratio of dietary linoleic acid to alpha-linolenic acid and BMD at the hip in 642 men, 564 women not using hormone therapy, and 326 women using hormone therapy; these results were independent of age, body mass index, and lifestyle factors. An increasing ratio of total dietary n-6 to n-3 fatty acids was also significantly and independently associated with lower BMD at the hip in all women and at the spine in women not using hormone therapy. CONCLUSIONS: A higher ratio of n-6 to n-3 fatty acids is associated with lower BMD at the hip in both sexes. These findings suggest that the relative amounts of dietary polyunsaturated fatty acids may play a vital role in preserving skeletal integrity in older age. PMID- 15817875 TI - Can the amount of Helicobacter pylori in the stomach be kept low through probiotic intake? PMID- 15817876 TI - The role of a low-glycemic-index diet in the management of obesity. PMID- 15817877 TI - Sodium content of processed foods: 1983-2004. PMID- 15817878 TI - Need to establish threshold soy protein intake for cholesterol reduction. PMID- 15817879 TI - Vitamin A equivalency estimates: understanding apparent differences. PMID- 15817880 TI - High-density lipoproteins prevent the oxidized low-density lipoprotein-induced epidermal [corrected] growth factor receptor activation and subsequent matrix metalloproteinase-2 upregulation. AB - OBJECTIVE: The atherogenic oxidized low-density lipoprotein (oxLDL) induces the formation of carbonyl-protein adducts and activates the epidermal [corrected] growth factor receptor (EGFR) signaling pathway, which is now regarded as a central element for signal transduction. We aimed to investigate whether and by which mechanism the anti-atherogenic high-density lipoprotein (HDL) prevents these effects of oxLDL. METHODS AND RESULTS: In vascular cultured cells, HDL and apolipoprotein A-I inhibit oxLDL-induced EGFR activation and subsequent signaling by acting through 2 separate mechanisms. First, HDL, like the aldehyde scavenger dinitrophenyl hydrazine, prevented the formation of oxLDL-induced carbonyl protein adducts and 4-hydroxynonenal (HNE)-EGFR adducts. Secondly, HDL enhanced the cellular antioxidant defenses by preventing (through a scavenger receptor class B-1 (SR-BI)-dependent mechanism) the increase of intracellular reactive oxygen species (ROS) and subsequent EGFR activation triggered by oxLDL or H2O2. A pharmacological approach suggests that this protective effect of HDL is independent of cellular glutathione level and glutathione peroxidase activity, but it requires catalase activity. Finally, we report that oxLDL upregulates both membrane type 1 (MT1)-matrix metalloproteinase-1 (MT1-MMP) and MMP-2 through an EGFR-dependent mechanism and that HDL inhibits these events. CONCLUSIONS: HDLs block in vitro oxLDL-induced EGFR signaling and subsequent MMP-2 activation by inhibiting carbonyl adducts formation and cellular oxidative stress. These effects of HDL may participate to reduce cell activation, excessive remodeling, and alteration of the vascular wall. PMID- 15817881 TI - Platelet degranulation is associated with progression of intima-media thickness of the common carotid artery in patients with diabetes mellitus type 2. AB - BACKGROUND: Platelets play a key role in atherogenesis and thromboembolic complications in patients with type 2 diabetes. METHODS AND RESULTS: We prospectively examined the relationship between systemic platelet activation and progression of carotid wall thickness within 1 year in 105 patients with type 2 diabetes. The intima-media thickness (IMT) of the common carotid artery was measured bilaterally at study entry and after 1 year. Platelet activation was assessed with the use of immunologic markers of platelet activation (CD62P, CD63, and CD40L) and flow cytometry. The prevalence for progression of atherosclerotic carotid disease in this population was 55.2%. We found that platelet degranulation (CD63 and CD40L) correlated with progression of IMT within 1 year (CD63: r=0.231, P=0.022; CD40L: r=0.230, P=0.029). Diabetic patients with progression of IMT had a significantly increased expression of CD63 compared with patients with stable carotid disease (mean intensity of immunofluorescence; median, interquartile range: 17.1 [12.4, 25.8] versus 11.9 [7.7, 19.8]; P=0.004). Multivariate logistic regression analysis revealed that degranulation of platelet CD63 is a predictor for progression of IMT independently of classical cardiovascular risk factors and hemoglobin A1c in diabetic patients (P=0.017). CONCLUSIONS: Enhanced systemic platelet degranulation is associated with progression of carotid artery disease in type 2 diabetes. PMID- 15817882 TI - Vascular implications of the Kruppel-like family of transcription factors. AB - The Kruppel-like factor (KLF) family is a recently highlighted group of zinc finger transcription factors given their important biological roles which include the vasculature. KLF2, KLF4, KLF5, and KLF6 are notable factors that have been implicated in developmental as well as pathological vascular processes. In this brief review, we provide an up-to-date summary of the physiological functions and cellular effects as well as transcriptional regulatory mechanisms of the vascular KLFs. Through such, we aim to provide a working view for understanding the pathological actions of KLFs in the vasculature. PMID- 15817883 TI - Long-term inhibition of RhoA attenuates vascular contractility by enhancing endothelial NO production in an intact rabbit mesenteric artery. AB - RhoA plays a critical role in regulating NO production in cultured endothelial cells. To determine its role in in situ endothelial cells, we investigated the effects of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase inhibitors and a RhoA binding domain of Rho-kinase (RB) on vascular contractility in the isolated rabbit mesenteric artery. Ex vivo treatment of the strips with 3x10(-5) mol/L simvastatin and fluvastatin for approximately 24 to 30 hours significantly attenuated the contractile response to phenylephrine and high K+ in the presence of endothelium. The addition of N(omega)-nitro-L-arginine methyl ester and the removal of endothelium abolished the attenuation of the contractile response. The cotreatment with geranylgeranyl pyrophosphate prevented the statin-induced attenuation of the contractile response, whereas geranylgeranyl transferase inhibitor mimicked the effect of simvastatin. Treatment with simvastatin enhanced the bradykinin-induced endothelium-dependent relaxation in the mesenteric artery, whereas it had no effect on the bradykinin-induced [Ca2+]i elevation in endothelial cells of the aortic valves. Introduction of RB to the strips using a cell-penetrating peptide of Tat protein (TATHA-RB) attenuated the contractile responses in a NO-dependent manner. However, a Rac1/Cdc42-binding fragment of p21 activated protein kinase, RB without Tat peptide or TATHA-protein A had no effect. The in vivo treatment of rabbit with simvastatin and TATHA-RB attenuated the contractility in a NO-dependent manner. Simvastatin and TATHA-RB significantly upregulated eNOS in the rabbit mesenteric artery. The present study provides the first evidence that RhoA plays a physiological role in suppressing NO production in in situ endothelial cells. PMID- 15817884 TI - Lack of myoglobin causes a switch in cardiac substrate selection. AB - Myoglobin is an important intracellular O2 binding hemoprotein in heart and skeletal muscle. Surprisingly, disruption of myoglobin in mice (myo-/-) resulted in no obvious phenotype and normal cardiac function was suggested to be mediated by structural alterations that tend to steepen the oxygen pressure gradient from capillary to mitochondria. Here we report that lack of myoglobin causes a biochemical shift in cardiac substrate utilization from fatty acid to glucose oxidation. Proteome and gene expression analysis uncovered key enzymes of mitochondrial beta-oxidation as well as the nuclear receptor PPAR to be downregulated in myoglobin-deficient hearts. Using FDG-PET we showed a substantially increased in vivo cardiac uptake of glucose in myo-/- mice (6.7+/ 2.3 versus 0.8+/-0.5% of injected dose in wild-type, n=5, P<0.001), which was associated with an upregulation of the glucose transporter GLUT4. The metabolic switch was confirmed by 13C NMR spetroscopic isotopomer studies of isolated hearts which revealed that [1,6-13C2]glucose utilization was increased in myo-/- hearts (38+/-8% versus 22+/-5% in wild-type, n=6, P<0.05), and concomitantly, [U 13C16]palmitate utilization was decreased in the myoglobin-deficient group (42+/ 6% versus 63+/-11% in wild-type, n=6, P<0.05). Because of the O2-sparing effect of glucose utilization, the observed shift in substrate metabolism benefits energy homoeostasis and therefore represents a molecular adaptation process allowing to compensate for lack of the cytosolic oxygen carrier myoglobin. Furthermore, our data suggest that an altered myoglobin level itself may be a critical determinant for substrate selection in the heart. The full text of this article is available online at http://circres.ahajournals.org. PMID- 15817885 TI - Reprogramming of the human atrial transcriptome in permanent atrial fibrillation: expression of a ventricular-like genomic signature. AB - Atrial fibrillation is associated with increased expression of ventricular myosin isoforms in atrial myocardium, regarded as part of a dedifferentiation process. Whether reexpression of ventricular isoforms in atrial fibrillation is restricted to transcripts encoding for contractile proteins is unknown. Therefore, this study compares atrial mRNA expression in patients with permanent atrial fibrillation to atrial mRNA expression in patients with sinus rhythm and to ventricular gene expression using Affymetrix U133 arrays. In atrial myocardium, we identified 1434 genes deregulated in atrial fibrillation, the majority of which, including key elements of calcium-dependent signaling pathways, displayed downregulation. Functional classification based on Gene Ontology provided the specific gene sets of the interdependent processes of structural, contractile, and electrophysiological remodeling. In addition, we demonstrate for the first time a prominent upregulation of transcripts involved in metabolic activities, suggesting an adaptive response to increased metabolic demand in fibrillating atrial myocardium. Ventricular-predominant genes were 5 times more likely to be upregulated in atrial fibrillation (174 genes upregulated, 35 genes downregulated), whereas atrial-specific transcripts were predominantly downregulated (56 genes upregulated, 564 genes downregulated). Overall, in fibrillating atrial myocardium, functional classes of genes characteristic of ventricular myocardium were found to be upregulated (eg, metabolic processes), whereas functional classes predominantly expressed in atrial myocardium were downregulated (eg, signal transduction and cell communication). Therefore, dedifferentiation with adoption of a ventricular-like signature is a general feature of the fibrillating atrium. PMID- 15817886 TI - Adenovirus-mediated overexpression of O-GlcNAcase improves contractile function in the diabetic heart. AB - To examine whether excessive protein O-GlcNAcylation plays a role in the dysfunction of the diabetic heart, we delivered adenovirus expressing O-GlcNAcase (Adv-GCA) into the myocardium of STZ-induced diabetic mice. Our results indicated that excessive cellular O-GlcNAcylation exists in the diabetic heart, and that in vivo GCA overexpression reduces overall cellular O-GlcNAcylation. Myocytes isolated from diabetic hearts receiving Adv-GCA exhibited improved calcium transients with a significantly shortened T(decay) (P<0.01) and increased sarcoplasmic reticulum Ca2+ load (P<0.01). These myocytes also demonstrated improved contractility including a significant increase in +dL/dt and -dL/dt and greater fractional shortening as measured by edge detection (P<0.01). In isolated perfused hearts, developed pressure and -dP/dt were significantly improved in diabetic hearts receiving Adv-GCA (P<0.05). These hearts also exhibited a 40% increase in SERCA2a expression. Phospholamban protein expression was reduced 50%, but the phosphorylated form was increased 2-fold in the diabetic hearts receiving Adv-GCA. We conclude that excess O-GlcNAcylation in the diabetic heart contributes to cardiac dysfunction, and reducing this excess cellular O GlcNAcylation has beneficial effects on calcium handling and diabetic cardiac function. PMID- 15817890 TI - Inhibition of NADPH oxidase attenuates vasospasm after experimental subarachnoid hemorrhage in rats. AB - BACKGROUND AND PURPOSE: The purpose of this study is to investigate whether apocynin, an NADPH oxidase inhibitor, attenuates vasospasm after experimental subarachnoid hemorrhage (SAH) in rats. METHODS: Rats were subjected to endovascular perforation of the right anterior cerebral artery or sham surgery. Beginning 2 hours after SAH, rats were administered 50 mg/kg apocynin or vehicle by intraperitoneal injection 3 times daily for 2 days. RESULTS: In SAH rats, apocynin treatment enlarged basilar artery diameter (SAH/apocynin=253+/-71 microm, SAH/saline=191+/-60 microm, P<0.01; SAH=190+/-58 microm, sham=276+/-52 microm, P<0.01), reduced neurological deficits (SAH/apocynin=24+/-6.5, SAH/saline=18+/-5.3, P<0.05; SAH=18+/-4.7, sham=27+/-0, P<0.01), decreased NADPH oxidase activity (SAH/apocynin=18.4+/-3.7, SAH/saline=25.7+/-5.2, P<0.05; SAH=27.5+/-5.8, sham=15.4+/-4.5 nmol/min per mg protein, P<0.05), decreased superoxide level (SAH/apocynin=6.5+/-1.8, SAH/saline=9.6+/-2.2, P<0.05; SAH=9.8+/ 1.9, sham=4.9+/-0.9 arbitrary units, P<0.05), and lowered membrane translocation of NADPH oxidase subunit p47phox. CONCLUSIONS: Inhibition of NADPH oxidase attenuates delayed cerebral vasospasm after experimental SAH, suggesting that the inhibition of NADPH oxidase may provide a therapeutic strategy for vasospasm after SAH. PMID- 15817891 TI - A digital map of middle cerebral artery infarcts associated with middle cerebral artery trunk and branch occlusion. AB - BACKGROUND AND PURPOSE: Knowledge of the topographic distribution of infarcts of the middle cerebral artery (MCA) may give insight into the limits of the arterial territory and infarct mechanism and may influence the decision to use thrombolytic therapy. We describe the creation of a digital atlas of MCA (DA-MCA) infarction associated with MCA branch and trunk occlusion using magnetic resonance (MR) techniques. METHODS: Hemispheric infarcts, with evidence of MCA trunk or branch occlusion, were manually segmented into binary images, linearly registered into a common stereotaxic coordinate space, and averaged to yield the probability of involvement by infarction at each voxel. Comparisons were made with existing maps of the MCA territory. RESULTS: Twenty-eight patients with median age of 74 years (range, 26 to 87 years) were studied. On the DA-MCA, the highest frequency of infarction was within the striatocapsular region, centrum semiovale, and the insula. The mean and maximal MCA infarct volumes were 195.5 cm3 and 366.3 cm3, respectively. Comparison with published maps showed that the most common difference from the DA-MCA was in the superomedial extent of the MCA territory. Some maps showed the MCA territory reaching the interhemispheric fissure, whereas in the DA-MCA it did not. There was a lower variability in the anterior boundary of the MCA territory compared with its posterior counterpart. CONCLUSIONS: We have created a digital atlas of MCA infarction using MR imaging techniques. This approach may be useful to establish the distribution of the MCA and other arterial territories and the border zones between them with greater certainty. PMID- 15817892 TI - Critical role of angiotensin II in excess salt-induced brain oxidative stress of stroke-prone spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: The detailed role of angiotensin II in salt-exacerbated stroke is unclear. We examined the role of angiotensin II in salt-accelerated stroke of stroke-prone spontaneously hypertensive rats (SHRSP). METHODS: Salt loaded SHRSP were orally given the angiotensin II type 1 (AT1) receptor blocker candesartan (0.3 to 3 mg/kg per day) and calcium channel blocker amlodipine (1 mg/kg per day), and the effects on stroke (n=61) and brain superoxide were compared between them. We also examined the effect of angiotensin II infusion (200 ng/kg per min) on brain superoxide production and blood-brain barrier. RESULTS: Despite the comparable hypotensive effect between candesartan and amlodipine, candesartan prolonged survival of salt-loaded SHRSP much more than amlodipine (P<0.01), being associated with more improvement of cerebral arteriolar thickening, cerebral arteriolar cell proliferation, and hippocampal CA1 neuronal cell reduction (1024.9+/-20.6 versus 724.9+/-22.8 cells/mm2; P<0.01; n=7 to 10 in each group) in SHRSP by candesartan (P<0.05) than amlodipine. Salt loading increased superoxide and NADPH oxidase activity in brain cortex and hippocampus of SHRSP, and this increase was prevented by candesartan (P<0.01) but not amlodipine. Angiotensin II infusion, via AT1 receptor, directly increased brain superoxide by 1.8-fold (P<0.05; n=6 to 7 in each group) and impaired blood brain barrier in salt-loaded SHRSP by 1.7-fold (P<0.05), and this increase in brain superoxide and blood-brain barrier impairment was prevented by tempol as well as candesartan. CONCLUSIONS: Excess salt, via oxidative stress, accelerates stroke, and angiotensin II, via AT1 receptor, plays a pivotal role in brain superoxide production of SHRSP by excess salt. PMID- 15817893 TI - Impaired cerebral vasoreactivity in a transgenic mouse model of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy arteriopathy. AB - BACKGROUND AND PURPOSE: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited small vessel disease causing stroke and dementia. The disease is caused by highly stereotyped mutations in NOTCH3, which is restrictively expressed in vascular smooth muscle cells (VSMCs). The mechanisms of compromised cerebral hemodynamics in CADASIL remain to be elucidated. We tested the hypothesis that mutant NOTCH3 impairs the vasomotor function of cerebral vessels. METHODS: Vasomotor function was examined in vivo in transgenic mice expressing a mutant NOTCH3 in VSMCs (TgNotch3R90C). Mice develop an age-dependent arteriopathy similar to that seen in CADASIL, without brain parenchyma lesions. Using laser-Doppler flowmetry, we assessed in awake TgNotch3R90C mice and wild-type littermates the cerebrovascular reactivity to 2 potent vasodilator stimuli (acetazolamide and hypercapnia) and cerebral blood flow (CBF) autoregulation during stepwise blood pressure elevations and reductions. Mice were studied at 18 months of age, when the CADASIL features are apparent, and at 10 months of age, before their appearance. RESULTS: Eighteen month-old TgNotch3R90C mice showed reduced responses to hypercapnia and acetazolamide, higher cerebrovascular resistance during hypertension, and their lower limit of CBF autoregulation was shifted to higher blood pressures. Cerebrovascular responses were similarly impaired in 10-month-old TgNotch3R90C mice. CONCLUSIONS: Cerebrovascular reactivity is compromised early in TgNotch3R90C mice. The data show an impaired autoregulation and are suggestive of a decreased relaxation or increased resistance of cerebral vessels. Our findings indicate that vascular dysfunction is an early pathogenic event that may promote the subsequent development of brain ischemia in CADASIL. PMID- 15817894 TI - Absolute stroke mortality burden for non-Hispanic non-Latino whites was disproportionately higher than expected simply based on the US population in 2001. AB - BACKGROUND AND PURPOSE: The absolute burden of stroke is a major determinant of health care costs and should also be considered when developing and implementing effective health policy. This study evaluated the impact of specific racial ethnic categorization on absolute stroke mortality burden and population percentages. METHODS: In this population-based analysis, 2001 US data was used to compute absolute values of population and stroke mortality burden for white and black, and other racial-ethnic groups. To test the effect of age-mix, values were age-adjusted using the 2000 US standard population. The z test statistic was computed and a 2-tailed P value of <0.05 was considered significant. RESULT: Whites comprised a majority of the 2001 absolute US stroke mortality burden and US population (86% and 82%, respectively). Surprisingly, nHnL whites comprised a much higher percentage of the absolute US stroke mortality burden than expected based on their percentage of the US population alone (81% and 69%, respectively; P<0.001). Age-adjustment indicated a contribution by age-mix, however, an age independent residual component remained. CONCLUSIONS: Specific race-ethnicity categorization significantly influences comparisons of the proportion of absolute stroke mortality burden to the population proportion. Accordingly, appropriate caution and care are needed when estimating the societal impact of conditions such as stroke. PMID- 15817895 TI - Oxygen glucose deprivation switches the transport of tPA across the blood-brain barrier from an LRP-dependent to an increased LRP-independent process. AB - BACKGROUND AND PURPOSE: Despite uncontroversial benefit from its thrombolytic activity, the documented neurotoxic effect of tissue plasminogen activator (tPA) raises an important issue: the current emergency stroke treatment might not be optimum if exogenous tPA can enter the brain and thus add to the deleterious effects of endogenous tPA within the cerebral parenchyma. Here, we aimed at determining whether vascular tPA crosses the blood-brain barrier (BBB) during cerebral ischemia, and if so, by which mechanism. METHODS: First, BBB permeability was assessed in vivo by measuring Evans Blue extravasation following intravenous injection at 0 or 3 hours after middle cerebral artery electrocoagulation in mice. Second, the passage of vascular tPA was investigated in an in vitro model of BBB, subjected or not to oxygen and glucose deprivation (OGD). RESULTS: We first demonstrated that after focal permanent ischemia in mice, the BBB remains impermeable to Evans Blue in the early phase (relative to the therapeutic window of tPA), whereas at later time points massive Evans Blue extravasation occurs. Then, the passage of tPA during these 2 phases, was investigated in vitro and we show that in control conditions, tPA crosses the intact BBB by a low-density lipoprotein (LDL) receptor-related protein (LRP) dependent transcytosis, whereas OGD leads to an exacerbation of tPA passage, which switches to a LRP-independent process. CONCLUSIONS: We evidence 2 different mechanisms through which vascular tPA can reach the brain parenchyma, depending on the state of the BBB. As discussed, these data show the importance of taking the side effects of blood-derived tPA into account and offer a basis to improve the current thrombolytic strategy. PMID- 15817896 TI - Screening for aspirin responsiveness after transient ischemic attack and stroke: comparison of 2 point-of-care platelet function tests with optical aggregometry. AB - BACKGROUND AND PURPOSE: Recent studies suggest that patients who do not respond to aspirin (ASA) therapy may be at increased risk of ischemic vascular events. The availability of simple to use point-of-care (POC) platelet function tests now potentially allows aspirin nonresponsiveness to be identified in routine clinical practice. However, there are very few data on whether the different tests produce consistent results. We therefore compared 2 POC tests (PFA-100 device and the Ultegra-RPFA [RPFA]) with conventional light transmission aggregometry (LTA). METHODS: Platelet function was assessed by all 3 tests in 100 patients receiving low-dose ASA therapy after transient ischemic attack (TIA) or ischemic stroke. RESULTS: The incidence of ASA nonresponsiveness was 17% by the RPFA and 22% by the PFA-100, compared with only 5% by LTA (ie, as defined with both arachidonic acid and ADP). Agreement between the RPFA and the PFA-100 and arachidonic acid induced LTA was poor (kappa=0.16, 95% CI, -0.08 to 0.39, P=0.11; and kappa=0.09 0.12 to 0.30, P=0.32, respectively). Agreement between the 2 POC tests was also poor (kappa=0.14, -0.08 to 0.36, P=0.15). Only 2% of patients were aspirin nonresponders by all 3 tests. CONCLUSIONS: The prevalence of apparent ASA nonresponsiveness was higher with both the POC tests than with LTA. However, agreement between the tests was poor and very few patients were ASA nonresponsive by all 3 tests. Aspirin nonresponsiveness is therefore highly test-specific and large prospective studies will be required to determine the prognostic value of each of the separate tests. PMID- 15817897 TI - Asymptomatic embolization detected by Doppler ultrasound predicts stroke risk in symptomatic carotid artery stenosis. AB - BACKGROUND AND PURPOSE: Asymptomatic cerebral emboli can be detected using transcranial Doppler ultrasound (TCD). These embolic signals have potential as a marker of stroke risk and as a surrogate marker to evaluate antiplatelet agents. Small studies have demonstrated that they predict the combined endpoint of stroke and transient ischemic attack (TIA), but no studies have shown that they predict the more important endpoint of stroke alone. METHODS: TCD was used to record for 1 hour from the ipsilateral middle cerebral artery in 200 patients with >50% symptomatic carotid stenosis. The Doppler audio signal was recorded for later analysis blinded to clinical details. Subjects were followed-up prospectively until surgical intervention, stroke, or study end at 90 days. RESULTS: Embolic signals (ES) were detected in 89 (44.5%). During follow-up, 31 subjects experienced recurrent ipsilateral ischemic events: 7 strokes and 24 TIAs. The presence of ES predicted stroke alone (P=0.001) and the combined endpoint of stroke and TIA (P=0.00001). This remained significant, with an odds ratio of 4.67 (95% CI, 1.99 to 11.01; P<0.0001) after Cox regression to control for age, sex, smoking, hypertension, time from last symptoms, and degree of stenosis. The absence of ES identified a group at low risk for stroke alone and stroke and TIA during follow-up: 0% and 7.5%, respectively, versus 3.5% and 15.5% in all 200 subjects. CONCLUSIONS: Asymptomatic embolization in carotid stenosis predicts short-term ipsilateral stroke risk. This supports use of the technique to identify patients at high-risk for recurrent stroke for therapeutic interventions and as a surrogate marker to evaluate antithrombotic medication. PMID- 15817898 TI - Development and validation of the stroke action test. AB - BACKGROUND AND PURPOSE: Accurately assessing the public's readiness to respond to stroke is important. Most published measures are based on recall or recognition of stroke symptoms, or knowledge of the best action for stroke when the diagnosis is provided. The purpose of this study was to develop and evaluate a new written instrument whose items require the respondent to associate individual symptoms with the most appropriate action. METHODS: The Stroke Action Test (STAT) contains 21 items that name or describe stroke symptoms from all 5 groups of warning signs and 7 items that are nonstroke symptoms. For each item, the respondent selects 1 of 4 options: call 911, call doctor, wait 1 hour, or wait 1 day. The instrument validation sample included 249 subjects from community-based organizations. Score reliability and validity were analyzed using multiple data and information sources. RESULTS: The mean overall STAT score (all 28 items) for the lay people was 36.8%. On average, they chose call 911 for 34.1% of the stroke symptoms. They chose call doctor for 39.4% of the stroke symptoms, wait 1 hour for 20.1%, and wait 1 day for 6.0%. Score reliability is good (alpha=0.83). Evidence confirming score validity is presented based on analysis of item content and response patterns, and examination of the relationships between test scores and key variables related to stroke knowledge. CONCLUSIONS: STAT directly assesses a critical aspect of practical stroke knowledge that has been largely overlooked and provides scores with good reliability and validity. PMID- 15817899 TI - The value of XYZ/2 technique compared with computer-assisted volumetric analysis to estimate the volume of chronic subdural hematoma. AB - BACKGROUND AND PURPOSE: A simple estimation method of intracerebral hematoma volume known as XYZ/2 method has been described previously. This method has also been shown to be valid for the estimation of acute subdural hematoma volume. However, chronic subdural hematomas differ in shape and extension from acute subdural hematomas, which makes the validity of the same method in the estimation of hematoma volume questionable. We aimed to determine the value of XYZ/2 method to estimate the volume of chronic subdural hematoma when compared with computer assisted volumetric analysis. METHODS: Computed tomography scans of 28 patients with unilateral hemispheric chronic subdural hematoma were reviewed. Hematoma volumes were measured using 5 different XYZ/2 formulas and compared with volumes measured by computer-assisted analysis. Nonparametric correlation coefficient (Spearman's rho) was used in statistical comparison. RESULTS: All 5 formulas showed excellent correlation with the gold standard, proving the validity of XYZ/2 method in the estimation of chronic subdural hematoma volume (level of significance <0.001). Our results suggest that maximum hematoma length and width, which are not necessarily on the same slice, should be used rather than length and width of hematoma on the central slice when using XYZ/2 method in patients with chronic subdural hematoma. CONCLUSIONS: This study proves the validity of XYZ/2 technique for the estimation of chronic subdural hematoma volume as well. PMID- 15817901 TI - Inter-religious perspectives on hope and limits in cancer treatment. PMID- 15817903 TI - Smoking and hormones in health and endocrine disorders. AB - Smoking has multiple effects on hormone secretion, some of which are associated with important clinical implications. These effects are mainly mediated by the pharmacological action of nicotine and also by toxins such as thiocyanate. Smoking affects pituitary, thyroid, adrenal, testicular and ovarian function, calcium metabolism and the action of insulin. The major salient clinical effects are the increased risk and severity of Graves' hyperthyroidism and opthalmopathy, osteoporosis and reduced fertility. Smoking also contributes to the development of insulin resistance and hence type 2 diabetes mellitus. An important concern is also the effect of smoking on the foetus and young children. Passive transfer of thiocyanate can cause disturbance of thyroid size and function. Furthermore, maternal smoking causes increased catecholamine production, which may contribute to under perfusion of the foetoplacental unit. PMID- 15817904 TI - Gonadal dysfunction in systemic diseases. AB - Gonadal function is significantly affected in many acute and chronic systemic diseases. As the function of the testes and the ovaries is determined by the integrity of the hypothalamic-pituitary-gonadal axis, it is obvious that a systemic disease may affect one or more levels of the axis in such a manner that the gonadal dysfunction may have various clinical and laboratory manifestations. In this brief review, the most common disturbances seen in the main systemic diseases will be discussed. PMID- 15817905 TI - Early manifestation of calcinosis cutis in pseudohypoparathyroidism type Ia associated with a novel mutation in the GNAS gene. AB - OBJECTIVE: To clarify the molecular defect for the clinical finding of congenital hypothyroidism combined with the manifestation of calcinosis cutis in infancy. CASE REPORT: The male patient presented with moderately elevated blood thyrotropin levels at neonatal screening combined with slightly decreased plasma thyroxine and tri-iodothyronine concentrations, necessitating thyroid hormone substitution 2 weeks after birth. At the age of 7 months calcinosis cutis was seen and the patient underwent further investigation. Typical features of Albright's hereditary osteodystrophy (AHO), including round face, obesity and delayed psychomotor development, were found. METHODS AND RESULTS: Laboratory investigation revealed a resistance to parathyroid hormone (PTH) with highly elevated PTH levels and a reduction in adenylyl cyclase-stimulating protein (Gsalpha) activity leading to the diagnosis of pseudohypoparathyroidism type Ia (PHP Ia). A novel heterozygous mutation (c364T > G in exon 5, leading to the amino acid substitution Ile-106 --> Ser) was detected in the GNAS gene of the patient. This mutation was not found in the patient's parents, both of whom showed normal Gsalpha protein activity in erythrocytes and no features of AHO. A de novo mutation is therefore likely. CONCLUSIONS: Subcutaneous calcifications in infancy should prompt the clinician to a thorough search for an underlying disease. The possibility of AHO and PHP Ia should be considered in children with hypothyroidism and calcinosis cutis. Systematic reviews regarding the frequency of calcinosis in AHO are warranted. PMID- 15817906 TI - [18F]fluoro-2-deoxy-D-glucose ([18F]FDG) positron emission tomography imaging of thymic carcinoid tumor presenting with recurrent Cushing's syndrome. AB - We report a case of a young woman with Cushing's syndrome (CS), in whom although endocrine investigations and negative pituitary imaging were suggestive of ectopic ACTH secretion, the results of inferior petrosal sinus (IPS) sampling after coricotropin-releasing hormone (CRH) stimulation were suggestive of pituitary ACTH hypersecretion. (111)In-labelled octreotide and high-resolution computer tomography (CT) revealed a lesion possibly responsible for the ACTH source in the thymus. Thymectomy confirmed concomitant ectopic CRH and probable ACTH production by a thymic neuroendocrine carcinoma. After an 8-year remission period the patient developed a clinical and biochemical relapse. A high resolution computed tomography (CT) scan of the thorax showed a 2-cm nodule in the thymic bed, which was positive on a [(18)F]fluoro-2-deoxy-D-glucose ([(18)F]FDG) positron emission tomography (PET) scan. However, a repeated thymectomy did not result in remission. A repeat [(18)F]FDG PET study showed persistent disease in the thymic bed and also uptake in the adrenals. The patient underwent bilateral adrenalectomy, which resulted in clinical remission. A further [(18)F]FDG PET scan 8 months later showed no progression of the thymic tumor and confirmed complete excision of the adrenals. This is a rare case of concomitant CRH and ACTH secretion from a thymic carcinoid tumor; the case illustrates the usefulness of functional imaging with [(18)F]FDG PET in the diagnosis, management and follow-up of neuroendocrine tumors. PMID- 15817907 TI - Cardiac function in borderline hypothyroidism: a study by pulsed wave tissue Doppler imaging. AB - OBJECTIVE: In subclinical hypothyroidism (SH), impaired diastolic function has been documented at rest and on effort, while systolic dysfunction has only been assessed on effort. DESIGN: The aim of the present study was: (a) to further assess systolic function at rest in SH; and (b) to ascertain whether cardiac dysfunction could precede TSH increase in euthyroid patients with a high risk of developing SH. METHODS: We studied 32 patients with classical Hashimoto's thyroiditis (22 with increased serum TSH (> 3 mU/ml - group A), and 10 with normal serum TSH (< 3 mU/ml - group B)); a third group (C), which included 13 healthy controls. All subjects underwent pulsed wave tissue Doppler imaging (PWTDI) to accurately quantify the global and regional left ventricular function. RESULTS: When compared with group C, PWTDI indices showed that in both groups A and B there was a significant impairment of systolic ejection (P < 0.001 and P < 0.05, respectively), a delay in diastolic relaxation (P < 0.001 and P < 0.05, respectively) and a decrease in the compliance to the ventricular filling (P < 0.05). Several significant correlations were found between PWTDI parameters and serum-free T(3) and T(4) and TSH concentrations. CONCLUSION: PWTDI is a sensitive technique that allows detection of both diastolic and systolic abnormalities, not only in patients with SH, but also in euthyroid subjects with a high risk of developing thyroid failure. Futhermore, the significant correlations of several PWTDI indices with serum FT(3) and TSH concentrations strongly support the concept of a continuum spectrum of a slight thyroid failure in autoimmune thyroiditis extending to subjects with serum TSH still within the normal range. PMID- 15817908 TI - Increased risk of affective disorder following hospitalisation with hyperthyroidism - a register-based study. AB - OBJECTIVE: The pattern of comorbidity between thyroid disorders and affective disorder is not fully known. We assessed whether hospitalisation with hyperthyroidism was a risk factor for hospitalisation with affective disorder and evaluated the temporal relationship between these events. DESIGN: A historical prospective cohort study comparing patients with hyperthyroidism with patients with non-toxic goitre or osteoarthritis, using existing data from Danish registers. The observational period was from 1 January 1977 to 31 December 1999. METHODS: Three study cohorts were identified by their International Classification of Diseases (ICD) diagnoses at discharge from hospital and consisted of all patients with a first hospital admission with the index diagnoses of hyperthyroidism, osteoarthritis, or non-toxic goitre. Later admissions to psychiatric hospital wards with discharge ICD diagnoses of affective disorder were used as events of interest. Rates of re-admission were estimated using competing risks models in survival analyses. Age, sex, substance abuse, and calendar time were included as co-variables. RESULTS: A study sample of 183 647 patients discharged with an index diagnosis was identified. In total 1374 events occurred in the observational period. An index diagnosis of hyperthyroidism was associated with an increased risk of hospitalisation with affective disorder for both sexes and for all age-bands investigated, compared with the other index diagnoses. The risk was greatest in the first six months after index hospitalisation (rate ratio, 95% confidence interval: 3.60 (2.58 5.04)). CONCLUSIONS: Patients hospitalised with hyperthyroidism are at greater risk of re-admission with depressive disorder or bipolar disorder than control patients. This suggests that hyperthyroidism is associated with long-term mood disturbances. PMID- 15817909 TI - High prevalence of cyclooxygenase 2 expression in papillary thyroid carcinoma. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) seems to play a role in the development and carcinogenesis of papillary thyroid carcinoma. Its incidence of expression and potential application as a tumor marker remain to be elucidated. MATERIALS AND METHODS: Immunohistochemical staining for COX-2 expression was performed for 30 papillary thyroid carcinoma (PTC) and 40 benign thyroid specimens. COX-2 mRNA expression was analyzed using a reverse transcriptase-polymerase chain reaction (RT-PCR) for paired fresh frozen tissues removed from surgically resected PTC specimens. RESULTS: COX-2 expression was detected by immunohistochemistry in 27 of 30 (90%) PTC but was absent in 40 benign thyroid specimens, including 27 nodular hyperplasia, 7 follicular adenoma and 6 lymphocytic thyroiditis. Two of the three COX-2 negative carcinomas were follicular variant of PTC. RT-PCR analysis confirmed COX-2 mRNA over-expression in 14 of 20 (70%) paired specimens of PTC. Real-time quantitative RT-PCR showed that the level of COX-2 mRNA expression was significantly higher in PTC than in both the adjacent non cancerous tissues and the benign thyroid specimens. CONCLUSION: COX-2 is frequently expressed in PTC but not in benign thyroid specimens. COX-2 expression may serve as a useful molecular marker for PTC in cases of diagnostic difficulty. PMID- 15817910 TI - Nitric oxide mediates abnormal responsiveness of thyroid arteries in methimazole treated patients. AB - OBJECTIVE: We studied the intervention of nitric oxide (NO), prostacyclin and endothelium-derived hyperpolarizing factor (EDHF) in mediating responses to acetylcholine in thyroid arteries from euthyroid and methimazole-treated (MT) patients. DESIGN AND METHODS: Branches of the superior thyroid artery were obtained from 19 euthyroid patients and 17 MT patients (euthyroid at the time of surgery) undergoing total thyroidectomy or hemithyroidectomy. Artery rings were suspended in organ baths for isometric recording of tension. RESULTS AND CONCLUSIONS: Acetylcholine caused endothelium-dependent relaxation of greater magnitude in arteries from MT patients (pD(2) (-log EC(50)) values were 7.68 +/- 0.19 in euthyroid and 8.17 +/- 0.26 in MT patients, P < 0.05). The relaxation was unaffected by indomethacin and was partially reduced by the NO-synthase inhibitor NG-monomethyl-L-arginine (L-NMMA). This reduction was higher in arteries from MT patients (50 +/- 6%) as compared with euthyroid patients (36 +/- 6%) (P < 0.05). Inhibition of K(+) channels using apamin combined with charybdotoxin or high K(+) solution abolished the relaxation resistance to L-NMMA and indomethacin. The maximal contraction response to noradrenaline (as a percentage of the response to 100 mM KCl) was lower in MT than in euthyroid patients (57 +/- 10 and 96 +/- 8 respectively, P < 0.05). The hyporesponsiveness to noradrenaline in arteries from MT patients was corrected by L-NMMA. The results indicate that: (i) thyroid arteries from MT patients show an increased relaxation response to acethylcholine and a decreased contraction response to noradrenaline due to overproduction of NO; (ii) EDHF plays a prominent role in acetylcholine-induced relaxation through activation of Ca(2+)-activated K(+) channels; (iii) the abnormal endothelium dependent responses in arteries from MT patients are not corrected by medical treatment. PMID- 15817911 TI - The clinical, metabolic and endocrine features and the quality of life in adults with childhood-onset craniopharyngioma compared with adult-onset craniopharyngioma. AB - BACKGROUND: Craniopharyngioma is a parasellar tumour that, although benign, tends to behave aggressively. It can occur at any age but most commonly presents in childhood or adolescence. OBJECTIVES: To investigate the frequency and severity of problems associated with craniopharyngioma, using the large international database (KIMS) for adult patients with GH deficiency (GHD), and to assess the differences between the adult onset (AO, aged 18 or above) disease and adults with childhood onset (CO) craniopharyngioma. DESIGN: Inclusion criteria were: an established diagnosis of craniopharyngioma, severe GHD and no recent GH treatment. These criteria were fulfilled by 393 (184 female, 209 male) patients; 241 had AO (mean age 28.7 +/- 8.7 years) and 152 had CO disease (age 42.0 +/- 12.3 years). Disease history, clinical features and anthropometric data were recorded at the time of enrolment in the database, and body composition, serum IGF-I, serum lipids and quality of life (QoL) were assessed. RESULTS: Peak age at onset of craniopharyngioma was 15-20 years. Ninety percent of patients had been treated surgically. CO patients were shorter than AO patients and had much lower IGF-I standard deviation scores (SDS). The majority had hypopituitarism and over 60% had diabetes insipidus. Body mass index (BMI) was higher in AO males (30.2 +/ 5.5) than in CO males (28.5 +/- 7.5); waist circumference was also greater. Obesity was more common in AO patients (51.8% vs 39.1%). Body composition did not differ between groups. Cholesterol and triglycerides were higher in AO than in CO patients, but high density lipoprotein (HDL)- and low density lipoprotein (LDL) cholesterol did not differ. Quality of life, assessed by Quality of Life Assessment of Growth Hormone Deficiency in Adults (QoL-AGHDA) and the Nottingham Health Profile, was markedly reduced in all groups with no significant differences between them; the QoL-AGHDA score correlated with age at onset of both craniopharyngioma and GHD, and also with BMI in AO patients. CONCLUSIONS: These data emphasise the generally poor state of health of patients treated for craniopharyngioma, with respect to endocrine and metabolic function, and also the markedly reduced quality of life. In addition to GHD, most patients have evidence of hypothalamic damage with associated obesity, diabetes insipidus and hypopituitarism. Adults with CO craniopharyngioma were shorter, had lower IGF-I, lower BMI, less obesity and slightly lower blood lipid levels than patients with AO craniopharyngioma. PMID- 15817912 TI - Effectiveness of adding dopamine agonist therapy to long-acting somatostatin analogues in the management of acromegaly. AB - BACKGROUND: The excess mortality and morbidity associated with acromegaly are secondary to prolonged elevation of GH and IGF-I. Vigorous control of these biochemical parameters results in improved morbidity and mortality. Somatostatin analogues (SAs) allow adequate control of GH and IGF-I in approximately 65% of subjects, leaving a significant cohort uncontrolled. Dopamine agonists (DAs), a cheap alternative to SAs, allow control of GH and IGF-I in less than 20% of patients with acromegaly. AIMS: To assess the effectiveness of adding DA therapy to SA in the biochemical control of acromegaly. SUBJECTS: One hundred and twenty cases from the Sheffield Acromegaly Register were reviewed; 24 (20%) did not require medical treatment following pituitary surgery alone; 16 (13%) had safe GH levels following surgery and radiotherapy; and 58 (48%) required medical treatment despite having had surgery, radiotherapy or both. The remaining 22 (18%) received only medical treatment. METHODS: In nine subjects a DA (three bromocriptine, six cabergoline) was added to an SA to control active disease. GH day curves and IGF-I levels were compared before and after the addition of a DA to existing SA treatment. All were on stable maximum-dose treatment with an SA, with inadequate biochemical control prior to addition of DA therapy. Mean duration of treatment on a DA before biochemical assessments were made was 10.3 months. Six subjects had previously been treated with either transsphenoidal surgery, radiotherapy or both. In three subjects SA was the primary therapy. RESULTS: All subjects exhibited a fall in median GH and IGF-I levels. Introduction of a DA resulted in a 36.1% reduction in median GH levels (8.3 vs 5.3 mIU/l; P = 0.008) on a GH day curve and a 35.2% reduction in IGF-I levels (387.2 vs 251.0 microg/l; P = 0.018). Only four subjects had elevated prolactin levels prior to the addition of a DA (>368 mIU/l). CONCLUSION: Addition of DAs to SAs is of benefit in the biochemical control of acromegaly and should be considered in those inadequately controlled. Furthermore, the beneficial effects of DAs occur even when pre-treatment prolactin levels are within the normal range. PMID- 15817913 TI - The GHRH/GHRP-6 test for the diagnosis of GH deficiency in elderly or severely obese men. AB - OBJECTIVE AND DESIGN: Ageing and obesity result in decreased activity of the GH/IGF-I axis and concomitant impaired GH responses to secretory stimuli. We therefore determined the validity of the GH cut-off value of 15.0 microg/l in the GH-releasing hormone (GHRH)/GH releasing peptide-6 (GHRP-6) test for the diagnosis of GH deficiency in elderly or severely obese men. METHODS: We performed a combined GHRH/GHRP-6 test in ten elderly men (mean age 74 years; mean body mass index (BMI) 24.6 kg/m(2)), nine obese men (mean age 47 years; mean BMI 40.6 kg/m(2)) and seven healthy male controls (mean age 51 years, mean BMI 24.3 kg/m(2)). After assessment of fasting plasma GH, IGF-I and IGF-binding protein-3 (IGFBP-3), GHRH (100 microg) and GHRP-6 (93 microg) were given intravenously as a bolus injection. Repeated GH measurements were performed for two hours. RESULTS: Both peak GH levels and areas under the curve (AUC) were significantly lower in the obese than in the controls (peak 13.2 vs 53.4 microg/l, P = 0.001; AUC 707 vs 3250 microg/l x 120 min; P = 0.001). Mean GH response in the elderly was lower than in the controls (peak 35.0 microg/l; AUC 2274 microg/l x 120 min), but this was not statistically significant. In contrast, GH peak levels in seven obese men remained below the cut-off level of 15.0 microg/l associated with severe GH deficiency. All others had GH peak levels exceeding this threshold. IGFBP-3 levels were significantly lower in the elderly than in the controls (1.35 vs 2.05 mg/l, P = 0.001). Baseline GH or IGF-I did not differ significantly between groups. CONCLUSIONS: GH responses following GHRH/GHRP-6 administration were significantly reduced in severely obese men, but were not significantly reduced in elderly men, despite a negative trend. Our data indicate that the cut-off GH level of 15.0 microg/l after GHRH + GHRP-6 administration for the diagnosis of severe GH deficiency cannot be used in severely obese men. PMID- 15817914 TI - Sheehan's syndrome: baseline characteristics and effect of 2 years of growth hormone replacement therapy in 91 patients in KIMS - Pfizer International Metabolic Database. AB - OBJECTIVE: Sheehan's syndrome occurs as a result of ischaemic pituitary necrosis due to severe postpartum haemorrhage. It is one of the most important causes of hypopituitarism, and hence growth hormone deficiency (GHD), in developing countries. However, little is known about the effects of growth hormone (GH) replacement therapy in patients with Sheehan's syndrome. DESIGN: The demographic background characteristics of 91 GH-deficient patients with Sheehan's syndrome (mean age +/- s.d., 46.3 +/- 9.4 years) were compared with those of a group of 156 GH-deficient women (mean age +/- s.d., 51.5 +/- 13.1 years) with a non functional pituitary adenoma (NFPA). The baseline characteristics and the effects of 2 years of GH replacement therapy were also studied in the 91 patients with Sheehan's syndrome and an age-matched group of 100 women with NFPA (mean age +/- s.d. 44.5 +/- 10.2 years). RESULTS: All patients were enrolled in KIMS (Pfizer International Metabolic Database). Patients with Sheehan's syndrome were significantly younger at pituitary disorder onset, diagnosis of GHD and at entry into KIMS than patients with NFPA (P < 0.01), and had significantly lower insulin like growth factor I levels (P < 0.001). At baseline, quality of life (QoL) was significantly (P < 0.05) reduced in patients with Sheehan's syndrome compared with those with NFPA (P < 0.001). With regard to treatment effects, lean body mass increased significantly (P < 0.05), QoL improved significantly (P < 0.05) and total and low-density lipoprotein-cholesterol decreased significantly (P < 0.05) in patients with Sheehan's syndrome after 1 year of GH replacement therapy. Similar significant changes in QoL and lipid profiles occurred in patients with NFPA after 2 years of GH replacement. Blood pressure remained unchanged in patients with Sheehan's syndrome, but decreased significantly (P < 0.01) in the group with NFPA after 1 year, before returning to pretreatment levels at 2 years. CONCLUSIONS: In conclusion, patients with Sheehan's syndrome have more severe GHD compared with individuals with NFPA. GH replacement therapy in patients with Sheehan's syndrome may have beneficial effects on QoL, body composition and lipid profile. PMID- 15817915 TI - Diagnosis of GH deficiency in the transition period: accuracy of insulin tolerance test and insulin-like growth factor-I measurement. AB - OBJECTIVE: A consensus exists that severe growth hormone deficiency (GHD) in adults is defined by a peak GH response to insulin-induced hypoglycemia (insulin tolerance test, ITT) of less than 3 microg/l based on a cohort of subjects with a mean age of 45 years. DESIGN AND METHODS: By considering one of the following two criteria for the diagnosis of probable permanent GHD, i.e. the severity of GHD (suggested by the presence of multiple pituitary hormone deficiencies (MPHD)) or the magnetic resonance (MR) imaging identification of structural hypothalamic pituitary abnormalities, 26 patients (17 males, 9 females, mean age 20.8 +/- 2.3 years, range 17-25 years) were selected for re-evaluation of the GH response to ITT and their IGF-I concentration. Eight subjects had isolated GHD (IGHD) and 18 had MPHD. Normative data for peak GH were obtained after ITT in 39 healthy subjects (mean age 21.2 +/- 4.4 years, range 15.1-30.0 years) and the reference range for IGF-I was calculated using normative data from 117 healthy individuals. RESULTS: Mean peak GH response to ITT was significantly lower in the 26 patients (1.8+/-2.0 microg/l, range 0.1-6.1 microg/l) compared with the 39 controls (18.5 +/- 15.5 microg/l, range 6.1-84.0 microg/l; P < 0.0001). One subject with septo optic dysplasia had a peak GH response of 6.1 microg/l that overlapped the lowest peak GH response obtained in normal subjects. There was an overlap for IGF-I SDS between subjects with IGHD and MPHD, as well as with normal controls. The diagnostic accuracy of a peak GH response of 6.1 microg/l showed a 96% sensitivity with 100% specificity. The maximum diagnostic accuracy with IGF-I SDS was obtained with a cut-off of -1.7 SDS (sensitivity 77%, specificity 100%) while an IGF-I < or = - 2.0 SDS showed a sensitivity of 62%. CONCLUSION: Our data show that the cut-off value of the peak GH response to ITT of less than 3 microg/l or 5 microg/l and of IGF-I of less than -2.0 SDS are too restrictive for the diagnosis of permanent GH deficiency in the transition period. We suggest that permanent GHD could be investigated more accurately by means of an integrated analysis of clinical history, the presence of MPHD, IGF-I concentration and the MR imaging findings of structural hypothalamic-pituitary abnormalities. PMID- 15817917 TI - Mini-Mental State Examination is superior to plasma glucose concentrations in monitoring patients with suspected hypoglycaemic disorders during the 72-hour fast. AB - OBJECTIVE: To determine whether systematic evaluation of cognitive function by the Mini-Mental State Examination (MMSE) allows the objective detection and documentation of cognitive deterioration in patients referred for evaluation of suspected hypoglycaemic disorders by the 72-h fast. DESIGN: Prospective case series. METHODS: In 50 patients referred for evaluation of suspected hypoglycaemic disorders, the MMSE score (maximum 30 points) was assessed at the start and at the end of the fast. RESULTS: The fast was terminated before 72 h in 14 patients because they developed neuroglycopenic symptoms due to hypoglycaemic disorders. Their MMSE score fell from a median of 29 points (range 20-30) at the beginning to 17 points (range 0-24) at the termination of the fast. The score dropped by > or =6 points in all patients with hypoglycaemic disorders. Median (range) plasma glucose concentration at the end of the fast was 2.1 (1.1-2.5) mmol/l. Thirty-six individuals developed no neuroglycopenic symptoms throughout the 72-h fast, their MMSE score remained between 27 and 30 throughout the fast and their median plasma glucose concentration dropped to 2.9 (2-3.6) mmol/l. CONCLUSIONS: Systematic evaluation of cognitive function by the MMSE at the beginning and at the termination of the fast allows objective determination and documentation of the deterioration of the cognitive state in patients with hypoglycaemic disorders. A decline in the cognitive performance by > or =6 points in the MMSE score rather than a distinct plasma glucose concentration should be used as the criterion to terminate the prolonged fast before 72 h. PMID- 15817916 TI - Ethinylestradiol and testosterone have divergent effects on circulating IGF system components in adolescents with constitutional tall stature. AB - OBJECTIVE: Pharmacological doses of estrogens or testosterone are used to limit the final height of girls or boys with constitutional tall stature but the mechanism behind this growth inhibition is still debated. We therefore studied the changes in the circulating components of the insulin-like growth factor (IGF) system during high dose sex steroid therapy. DESIGN AND METHODS: Twenty three girls and twenty boys with constitutional tall stature were treated with 100 microg ethinylestradiol per day or 250 mg testosterone ester every 14 days respectively. In 19 girls and 18 boys, the levels of IGF-I, free IGF-I, IGF-II, acid-labile subunit (ALS) and IGF binding proteins (IGFBP)-2 to -6 were measured before and 3-6 months after the start of therapy (group 1). In 18 girls and 11 boys, samples were collected at the end of therapy and 3 to 6 months afterwards (group 2). Fourteen girls and nine boys belonged to both groups. All parameters were measured by radioimmunoassay or ELISA. RESULTS: Levels of IGF-I were decreased significantly by estrogen treatment but remained unchanged during testosterone treatment. Free IGF-I decreased during estrogen treatment but increased during testosterone therapy. Estrogens increased IGF-II and testosterone reduced it. The important reduction of IGFBP-2 during estrogen therapy is not reproduced by androgen therapy, neither is the stimulation by estrogens of IGFBP-4. IGFBP-3 is not modulated by either sex steroid. We found that IGFBP-6 is up-regulated by testosterone but not by estrogens; the reverse is true for ALS, which increased during estrogen treatment but remained unchanged during testosterone treatment. CONCLUSIONS: Our findings demonstrate that androgens and estrogens exert differential effects on the circulating levels of several IGF components. PMID- 15817918 TI - Does metformin modify ovarian responsiveness during exogenous FSH ovulation induction in normogonadotrophic anovulation? A placebo-controlled double-blind assessment. AB - OBJECTIVE: To assess whether the addition of metformin to gonadotrophin ovulation induction in insulin-resistant, normogonadotrophic, anovulatory women alters ovarian responsiveness to exogenous FSH. DESIGN: Placebo-controlled double-blind assessment in an academic hospital. RESULTS: After a progestagen withdrawal bleeding, patients were randomised for either metformin (n = 11) or placebo (n = 9) treatment. In cases of absent ovulation, exogenous FSH was subsequently administered to induce ovulation. Only during metformin treatment did body mass index and androgen (androstenedione and testosterone) levels decrease, whereas FSH and LH levels increased significantly. In the metformin group, a single patient ovulated before the initiation of exogenous FSH. Significantly more monofollicular cycles and lower preovulatory oestradiol concentrations were observed in women receiving FSH with metformin compared with FSH alone. CONCLUSIONS: Metformin co-treatment in a group of insulin-resistant, normogonadotrophic, anovulatory patients resulted in normalization of the endocrine profile and facilitated monofollicular development during the FSH induction of ovulation. PMID- 15817919 TI - Forskolin and dexamethasone synergistically induce aromatase (CYP19) expression in the human osteoblastic cell line SV-HFO. AB - OBJECTIVE: Recent progress supports the importance of local estrogen secretion in human bone tissues to increase and maintain bone mineral density. In a previous study, we reported that the expression of aromatase (CYP19) is dexamethasone (Dex)-dependent and oncostatin M (OSM) increases the expression synergistically with Dex. In the present study, we examined the effects of forskolin (FSK) as another potential synergistic factor. RESULTS: Co-administration of 100 nM Dex and 10 microM FSK increased aromatase activity 4-fold compared with Dex alone. The results of reverse transcriptase (RT)-PCR suggest that the amount of CYP19 gene transcript was also up-regulated by FSK synergistically with Dex, and that promoter I.4, which is not activated by FSK alone, is activated by FSK synergistically with Dex. The results of RT-PCR also suggest that promoter II, which responds to FSK, was not activated even in the presence of FSK in SV-HFO. The promoter I.4 sequence that was transfected into SV-HFO was activated by FSK synergistically with Dex. CONCLUSIONS: Synergistic up-regulation of aromatase activity, CYP19 gene transcript, and promoter I.4 activity were Dex-dependent and not up-regulated by FSK alone. The results of this work may form the basis of bone-specific estrogen-replacement therapy that increases the estrogen concentration in bone tissue only. PMID- 15817920 TI - Interactions between the extracellular domain and the extracellular loops as well as the 6th transmembrane domain are necessary for TSH receptor activation. AB - OBJECTIVE: The molecular mechanisms of TSH receptor (TSHR) activation and intramolecular signal transduction are largely unknown. Deletion of the extracellular domain (ECD) of the TSHR results in increased constitutive activity, which suggests a self-inhibitory interaction between the ECD and the extracellular loops (ECLs) or the transmembrane domains (TMDs). To investigate these potential interactions and to pursue the idea that mutations in the ECD affect the constitutive activity of mutants in the ECLs or TMDs we generated double mutants between position 281 in the ECD and mutants in all three ECLs as well as the 6th TMD. DESIGN: We combined mutation S281D, characterized by an impaired TSH-stimulated cAMP response, with the constitutively activating in vivo mutations I486F (1st ECL), I568T (2nd ECL), V656F (3rd ECL) and D633F (6th TMD). Further, we constructed double mutants containing the constitutively activating mutation S281N and one of the inactivating mutations D474E, T477I (1st ECL) and D633K (6th TMD). RESULTS: The cAMP level of the double mutants with S281N and the inactive mutants in the 1st ECL was decreased below the level of the inactive single mutants, demonstrating that a constitutively activating mutation in the ECD cannot bypass disruption of signal transduction in the serpentine domain. In double mutants with S281D, basal and TSH-induced cAMP and inositol phosphate production of constitutively active mutants was reduced to the level of S281D. CONCLUSION: The dominance of S281D and the dependence of constitutively activating mutations in the ECLs on the functionally intact ECD strongly suggest that interactions between these receptor domains are required for TSHR activation and intramolecular signal transduction. PMID- 15817921 TI - CXCR6 within T-helper (Th) and T-cytotoxic (Tc) type 1 lymphocytes in Graves' disease (GD). AB - OBJECTIVE: In Graves' disease (GD), stimulating anti-TSH receptor antibodies are responsible for hyperthyroidism. T-helper 2 (Th2) cells were expected to be involved in the underlying immune mechanism, although this is still controversial. The aim of this study was to examine the expression of CXCR6, a chemokine receptor that marks functionally specialized T-cells within the Th1 and T-cytotoxic 1 (Tc1) cell pool, to gain new insights into the running immune processes. METHODS: CXCR6 expression was examined on peripheral blood lymphocytes (PBLs) and thyroid-derived lymphocytes (TLs) of GD patients in flow cytometry. CXCR6 cDNA was quantified in thyroid tissues affected by GD (n = 16), Hashimoto's thyroiditis (HT; n = 2) and thyroid autonomy (TA; n = 11) using real-time reverse transcriptase PCR. RESULTS: The percentages of peripheral CXCR6(+) PBLs did not differ between GD and normal subjects. CXCR6 was expressed by small subsets of circulating T-cells and natural killer (NK) cells. CXCR6(+) cells were enriched in thyroid-derived T-cells compared with peripheral CD4(+) and CD8(+) T-cells in GD. The increase was evident within the Th1 (CD4(+) interferon-gamma(+) (IFN gamma(+))) and Tc1 (CD8(+)IFN-gamma(+)) subpopulation and CD8(+) granzyme A(+) T cells (cytotoxic effector type). Thyroid-derived fibro-blasts and thyrocytes were CXCR6(-). There was no significant difference between the CXCR6 mRNA levels in GD compared with HT and normal TA tissues. The lowest CXCR6 mRNA levels were obtained from thyroid nodules from TA patients and GD patients with low thyroid peroxidase autoantibody levels. CONCLUSIONS: CXCR6 was overexpressed in Th1 and Tc1 TLs compared with PBLs in GD. CXCR6 could be a marker for lymphocytes that have migrated into the thyroid and assist in the thyroid, independently of the bias of the underlying disease. PMID- 15817922 TI - The multi-ligand somatostatin analogue SOM230 inhibits ACTH secretion by cultured human corticotroph adenomas via somatostatin receptor type 5. AB - OBJECTIVE: Currently, there is no effective medical treatment for patients with pituitary-dependent Cushing's disease. A novel somatostatin (SS) analogue, named SOM230, with high binding affinity to SS receptor subtypes sst(1), sst(2), sst(3) and sst(5) was recently introduced. We compared the in vitro effects of the sst(2)-preferring SS analogue octreotide (OCT) and the multi-ligand SOM230 on ACTH release by human and mouse corticotroph tumour cells. METHODS: By quantitative RT-PCR the sst subtype expression level was determined in human corticotroph adenomas. In vitro, the inhibitory effect of OCT and SOM230 on ACTH release by dispersed human corticotroph adenoma cells and mouse AtT20 corticotroph adenoma cells was determined. In addition, the influence of dexamethasone on the responsiveness to OCT and SOM230 was studied. RESULTS: Corticotroph adenomas expressed predominantly sst(5) mRNA (six out of six adenomas), whereas sst(2) mRNA expression was detected at significantly lower levels. In a 72 h incubation with 10 nmol/l SOM230, ACTH release was inhibited in three out of five cultures (range -30 to -40%). Ten nmol/l OCT slightly inhibited ACTH release in only one of five cultures (- 28%). In AtT20 cells, expressing sst(2), sst(3) and sst(5), SOM230 inhibited ACTH secretion with high potency (IC(50) 0.2 nmol/l). Dexamethasone (10 nmol/l) pre-treatment did not influence the sensitivity of the cells to the inhibitory effect of SOM230, suggesting that sst(5) is relatively resistant to negative control by glucocorticoids. CONCLUSIONS: The selective expression of sst(5) receptors in corticotroph adenomas and the preferential inhibition of ACTH release by human corticotroph adenoma cells by SOM230 in vitro, suggest that SOM230 may have potential in the treatment of patients with pituitary-dependent Cushing's disease. PMID- 15817923 TI - Norethisterone enanthate has neither a direct effect on the testis nor on the epididymis: a study in adult male cynomolgus monkeys (Macaca fascicularis). AB - OBJECTIVE: Norethisterone enanthate (NETE) is evaluated in trials of hormonal male contraception. It has been speculated that progestins may exert their contraceptive effects not only by suppressing gonadotropins but also by direct effects on male organs. NETE was given to monkeys in which endogenous gonadotropin secretion was suppressed by a gonadotropin releasing hormone (GnRH) antagonist, and replaced by human follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG). If NETE has a direct effect on spermatogenesis and/or epididymal function, some changes in testicular histology, sperm motility and/or morphology should occur soon after exposure to NETE. METHODS: Fifteen adult intact male monkeys were grouped and treated for a 38-day period. Group I received GnRH antagonist, FSH, hCG and NETE while group II received a regime identical to group I without NETE and group III received only NETE and vehicle. Ejaculates, body weight, testicular biopsies and volume, and hormones were evaluated. RESULTS: There was a similar pattern of serum FSH and testosterone in groups I and II. Testicular volume and the proportion of tubuli exhibiting spermatids was significantly decreased in group III. There were no significant differences between group I and group II in any parameters measured. The forward progression of sperm was not affected by NETE treatment. The consistently low percentages of grade c sperm indicated no sign of hyperactivation. No changes in the gross morphology of the acrosome were detected. CONCLUSIONS: Short-term NETE treatment has neither a direct effect on the testis nor on the epididymis in this nonhuman primate model and its contraceptive effects appear to be exerted exclusively through gonadotropin suppression. PMID- 15817924 TI - Differential effects of all-trans-retinoic acid (RA) on Erk1/2 phosphorylation and cAMP accumulation in normal and malignant human prostate epithelial cells: Erk1/2 inhibition restores RA-induced decrease of cell growth in malignant prostate cells. AB - OBJECTIVE: All-trans-retinoic acid (RA) regulates cellular growth, differentiation and apoptosis in human prostate by binding to RA receptors. Non genomic retinoid effects on signal transduction kinases in the cytoplasm are also described in several cells but they are still unknown in prostate cells. METHODS: Using an epithelial cell line derived from normal human prostate (EPN), and normal (NPEC) and malignant (CPEC) epithelial primary cultures of human prostate, we have examined effects of RA on both extracellular signal-regulated kinase 1/2 (Erk1/2) and cAMP accumulation. Then we have verified the effect of the inhibition of Erk1/2 on RA-induced growth arrest and apoptosis in malignant cells. RESULTS: In NPEC and in EPN treated with RA for up to 24 h, Western blot analyses of Erk1/2 phosphorylation show that RA causes a rapid activation of Erk1/2 within 5 min, which is maintained for 30 min, followed by a return to basal levels. In CPEC, the activated phosphorylation levels persist up to 24 h. While basal cAMP levels are not affected by 30 min treatment with RA in both EPN and NPEC, levels are increased in CPEC. Forskolin-induced cAMP levels are decreased by RA in all cell types. CPEC were incubated for up to 96 h with RA with and without the inhibitor of Erk1/2, UO126. CPEC incubated with RA and UO126 for 72 h showed a significant arrest of cell growth and after 96 h apoptosis in 11% of cells. CONCLUSIONS: We show rapid effects of RA on cytoplasmic messenger pathways in human prostate, and that responses can differ between normal and malignant cells. The inhibition of these pathways could improve the efficiency of RA in prostate cancer growth control. PMID- 15817936 TI - IJMR special issue on HIV/AIDS: editorial overview. PMID- 15817925 TI - Glucose triggers protein kinase A-dependent insulin secretion in mouse pancreatic islets through activation of the K+ATP channel-dependent pathway. AB - OBJECTIVE: To assess the significance of protein kinase A (PKA) in glucose triggering of ATP-sensitive K(+) (K(+)(ATP)) channel-dependent insulin secretion and in glucose amplification of K(+)(ATP) channel-independent insulin secretion. METHODS: Insulin release from cultured perifused mouse pancreatic islets was determined by radioimmunoassay. RESULTS: In islets cultured at 5.5 mmol/l glucose, and then perifused in physiological Krebs-Ringer medium, the PKA inhibitors, H89 (10 micromol/l) and PKI 6-22 amide (30 micromol/l) did not inhibit glucose (16.7 mmol/l)-induced insulin secretion, but inhibited stimulation by the adenylyl cyclase activator, forskolin (10 micromol/l). In the presence of 60 mmol/l K(+) and 250 micromol/l diazoxide, which stimulates maximum Ca(2+) influx independently of K(+)(ATP) channels, H89 (10 micromol/l) inhibited Ca(2+)-evoked insulin secretion, but failed to prevent glucose amplification of K(+)(ATP) channel-independent insulin secretion. In the presence of 1 mmol/l ouabain and 250 micromol/l diazoxide, which cause modest Ca(2+) influx, glucose amplification of K(+)(ATP) channel-independent insulin secretion was observed without concomitant Ca(2+) stimulation of PKA activity. In islets cultured at 16.7 mmol/l glucose, glucose (16.7 mmol/l)-induced insulin secretion in physiological Krebs-Ringer medium was augmented and now inhibited by H89 (10 micromol/l), implicating that culture at 16.7 mmol/l glucose may increase Ca(2+) sensitive adenylyl cyclase activity and hence PKA activity. In accordance, Ca(2+) evoked insulin secretion at 60 mmol/l K(+) and 250 micromol/l diazoxide was improved, whereas glucose amplification of K(+)(ATP) channel-independent insulin secretion was unaffected. CONCLUSIONS: Glucose may activate PKA through triggering of the K(+)(ATP) channel-dependent pathway. Glucose amplification of K(+)(ATP) channel-independent insulin secretion, on the other hand, occurs by PKA independent mechanisms. PMID- 15817937 TI - TRIPS, patents & HIV/AIDS drugs. PMID- 15817938 TI - Detection of Enterocytozoon bieneusi (Microsporidia) by polymerase chain reaction (PCR) using species-specific primer in stool samples of HIV patients. AB - BACKGROUND & OBJECTIVE: Microsporidia are obligate intracellular parasites causing infections predominantly in immunocompromised patients. Enterocytozoon bieneusi is the most important microsporidian causing chronic diarrhoea in AIDS patients. The current method used for diagnosing the microsporidia spores is based on light microscopy using stained smears, which do not differentiate spores at species level. The present study was undertaken to detect microsporidia and confirm at species level (E. bieneusi) by PCR from stool samples of HIV positive patients. METHODS: During September 2002 to April 2003, stool samples from 153 HIV-positive patients (with chronic diarrhoea n = 105; without diarrhoea n=48) were collected and examined microscopically for microsporidia spores using modified Weber's chromotrope stain. Stool samples were subjected to PCR assay using species-specific primer EBIEFI/EBIER1, which amplifies small subunit ribosomal RNA (SSU rRNA) of this microsporidian RESULTS: A total of 10 HIV positive patients with chronic diarrhoea were positive for microsporidia by microscopic analysis and confirmed as Enterocytozoon bieneusi by PCR. No false positive results were observed. A diagnostic DNA fragment of 607 bp of the unique SSU rRNA was amplified from all samples infected with E. bieneusi. INTERPRETATION & CONCLUSION: The study revealed that polymerase chain reaction is a useful tool for accurate species identification of microsporidia in stool samples, which serves the benefit of treatment to the patients. PMID- 15817939 TI - Serum neopterin levels in HIV infected patients with & without tuberculosis. AB - BACKGROUND & OBJECTIVE: Three categories of prognostic markers are best documented as having significance in relation to prognosis of HIV infection. These include HIV viral load, CD4 T-cell levels and plasma levels of soluble markers of immune activation. The plasma activation markers, like neopterin, tumor necrosis factor alpha (TNF-alpha), interleukins etc., are products of cytokine activity and represent immunologic changes throughout the body. There is not much information available on serum neopterin estimation in patients infected with both HIV and tuberculosis (TB), though neopterin levels are known to be elevated in pulmonary TB patients. In this study we attempted to correlate neopterin levels with the presence of tuberculosis in HIV infected and uninfected individuals and studied the changes after antituberculosis treatment. METHODS: Serum neopterin concentrations were measured by high performance liquid chromatography (HPLC) in 25 HIV-seropositive (HIV-TB) and 10-seronegative (TB) patients with tuberculosis before, during and at the end of antituberculosis therapy (ATT). S-neo was also measured in 10 HIV-seropositive asymptomatic individuals and 10 healthy controls. The results were correlated with clinical, bacteriological and immunological status. RESULTS: All TB patients regardless of HIV status had elevated s-neo concentrations at diagnosis, which declined gradually during treatment. Patients with HIV/TB with CD4 counts < 200/mm(3) had the highest levels at baseline with a steep fall during treatment. The median level at the end of treatment was significantly higher in HIV/TB than in TB patients, despite clinical improvement and bacteriological clearance of Mycobacterium tuberculosis. HIV infected asymptomatic individuals had neopterin levels that were higher than healthy controls but lower than HIV-TB patients. INTERPRETATION & CONCLUSION: Serum neopterin levels are elevated in HIV-positive patients, with the highest levels in those with tuberculosis and CD4 counts < 200/mm(3). Though the levels decrease with anti tuberculosis therapy, persistently elevated levels indicate progressive HIV disease and a poor prognosis. PMID- 15817940 TI - Subtype & cytokine profiles of HIV infected individuals from south India. AB - BACKGROUND & OBJECTIVE: The global surveillance of human immunodeficiency virus (HIV) subtypes (clades) helps understand the global distribution and incidence of different HIV subtypes. As knowledge about subtypes circulating in an area is needed for developing a candidate vaccine, prevalence of the subtypes HIV-1 and HIV-2 were studied in south India. The profile of cytokines interleukin 10 (IL10) and interferon gamma (IFNgamma) in both types of infection were also analysed as these are considered indicators of disease progression. METHODS: Patients who belonged to the 4 south Indian States i.e. Tamil Nadu, Kerala, Karnataka and Andhra Pradesh were included. HIV-1 subtyping was carried out by the heteroduplex mobility analysis (HMA) while that of HIV-2 was done by direct sequencing. The quantitation of IFNgamma and IL-10 was carried out using commercial ELISA kits. RESULTS: Among the 82 HIV-1 infected individuals subtyped, 78 (95.1%) were subtype C while all 12 HIV-2 strains were subtype A. IL-10 concentration was significantly higher among HIV infected individuals compared to normal healthy controls. IFNgamma was significantly higher among symptomatic and AIDS groups compared to asymptomatic HIV-1 infected individuals. INTERPRETATION & CONCLUSION: HIV-1 subtype C and the HIV-2 subtype A are the major subtypes circulating in south India. The study showed a trend towards a shifting of the cytokine profile from Th1 to Th2/Th0 in HIV-1, HIV-2 infections, and HIV-1 and HIV-2 dual infected individuals as the disease progresses. This trend observed is not unlike that reported from the West, despite the difference in subtype profile. PMID- 15817941 TI - Drug resistance pattern of Mycobacterium tuberculosis in seropositive and seronegative HIV-TB patients in Pune, India. AB - BACKGROUND & OBJECTIVE: Tuberculosis is the commonest opportunistic disease in persons infected with human immunodeficiency virus (HIV). Emergence of drug resistant isolates of M. tuberculosis highlights the need for continuous monitoring of drug resistance to antituberculosis drugs. Considering the reported high prevalence of drug resistance in HIV infected tuberculosis patients, we studied the anti-tuberculosis drug resistance pattern of M. tuberculosis in HIV seropositive and seronegative tuberculosis patients in Pune, Maharashtra, India. METHODS: A total of 70 M. tuberculosis isolates, 30 from HIV seropositive and 40 from HIV seronegative tuberculosis patients with no previous history of anti tuberculosis treatment, were isolated from sputum samples on Lowenstein-Jensen (LJ) medium, confirmed by conventional biochemical tests and stored at -70 degrees C. They were revived by subculturing on LJ medium and tested for drug resistance against four first-line antitubercular drugs by BACTEC Mycobacterial growth indicator tube 960 (MGIT 960) system. RESULTS: Of the 30 isolates from HIV infected patients, 10 per cent were resistant to isoniazid (H), 6.6 per cent to streptomycin (S), 6.6 per cent to ethambutol (E) and 10 per cent were multi drug resistant (MDR). Of the 40 M. tuberculosis isolates from HIV uninfected individuals, 10 per cent were resistant to H, 2.5 per cent to S, 2.5 per cent to E, and 2.5 per cent isolates were MDR. INTERPRETATION & CONCLUSION: The prevalence of drug resistant M. tuberculosis isolates among HIV seropositive tuberculosis patients was similar to that of HIV seronegative TB patients, indicating HIV infection may not be associated with drug resistant tuberculosis. However, considering the results from other studies and a high prevalence of HIV TB infection in the country, monitoring of drug resistance in M. tuberculosis isolates needs prioritization to ensure success in national tuberculosis control programme. PMID- 15817942 TI - Immunopathogenesis of HIV infection. AB - Progression of HIV infection is largely dependent on the interaction between the viral factors and host factors. HIV primarily infects the CD4 lymphocytes in the body. It brings about the destruction of CD4 cells through multiple mechanisms including apoptosis. The loss of CD4 cell population ultimately leads to the inability of infected person to deal with opportunistic organisms. Host genetic factors such as HLA polymorphism and HIV co-receptor polymorphism may influence either susceptibility to infection or disease progression. Innate immune mechanisms may play a role in disease progression. However, adaptive immune response is the most critical component of immune system for control of HIV infection. HIV-specific CD4 helper response and HIV-specific CTL responses have clearly emerged as the most important host factors that may decide the rate of disease progression. However, the role of neutralizing antibodies still remains to be understood in context with the disease progression. One of the gray areas is the role of mucosal immune response in HIV infection. However, it is clear that it is not a single component but orchestrated action of different immune mechanisms will decide the outcome of HIV infection. The studies in persons exposed to HIV infection but who are uninfected and the long term non-progressors will be critical for understanding the immunopathogenesis of HIV infection. PMID- 15817943 TI - HAART & the molecular biology of AIDS dementia complex. AB - The era of highly active antiretroviral therapy (HAART) has led to a considerable decline in the HIV disease progression rates and HIV-1-related opportunistic infections especially in developed countries. Unfortunately, antiretroviral treatment for almost 90 per cent of the HIV-infected population is not available because of cost concerns. Although a number of studies have shown uniform impact of HAART on disease progression, its effect on treating HIV infection of the brain and its manifestations, such as AIDS dementia complex (ADC), remains unclear. Along with the reasons why AIDS dementia complex continues to be a problem in the era of HAART, this review also discusses the changes in ADC patterns with HAART and its relevance in developing countries such as India. In addition, an overview of various biological, molecular and therapeutic aspects that may influence HIV dementia (HIV-D) is provided. PMID- 15817944 TI - HIV-1 viral protein R (Vpr) & host cellular responses. AB - During infection of host cells by HIV-1, active host-pathogen interactions take place. The final balance between these interactions determines the efficiency of viral infection and subsequent disease progression. HIV-infected cells respond to viral invasion with various antiviral strategies such as innate, cellular and humoral immune antiviral defense mechanisms. On the other hand, the virus has also developed tactics to suppress these host cellular responses. Among the many viral offensive strategies, viral protein R (Vpr) plays a particularly active role. Vpr involved in nuclear transport of the viral pre-integration complex, activation of viral transcription, induction of cell cycle G2/M arrest and apoptosis of the host cells. However, specific roles of these Vpr activities in viral pathogenesis and their contribution to disease progression are not fully understood. HIV-1 defective for some or all of these Vpr activities have been associated with slow disease progression in some patients. With regard to the host responses to vpr gene expression, studies show that Vpr is specifically targeted by CD8 T-lymphocytes during acute viral infection and that the host innate immune response may also play a crucial role in suppressing the effects of Vpr on various cellular activities. The effect of host cellular responses to vpr gene expression and its roles in nuclear transport, cell cycle G2/M regulation and induction of apoptosis are discussed in this review. Strategies with potential application for future antiviral therapies directed at suppressing Vpr activities are described. PMID- 15817945 TI - Impact of genetic diversity of HIV-1 on diagnosis, antiretroviral therapy & vaccine development. AB - HIV-1 strains have diversified extensively through mutation and recombination since their initial transmission to human beings many decades ago in central Africa. The high error rate of HIV reverse transcriptase combined with the estimated in vivo HIV-1 replication rate of ten billion new virions each day leads to extraordinary genetic diversity of HIV. Twenty seven circulating genetic forms of the HIV-1 group M are presently recognized, including 11 subtypes and sub-subtypes, and 16 circulating recombinant forms (CRF). Genotypic analyses have provided a better understanding of the molecular diversity of HIV-1, enabling the detection of emerging HIV-1 variants and improving the tracking of the epidemic worldwide. The rapid evolution of HIV within infected hosts contributes significantly to the elusiveness of this pathogen from host antiviral responses. The complex nature of HIV envelope glycoprotein that is inherently resistant to neutralization, the selective infection, progressive destruction and impaired regeneration of CD4+ T helper cells, generation of cytotoxic T lymphocyte (CTL) escape mutants, together with high genetic diversity with continually evolving HIV variants worldwide, makes design of an effective vaccine a formidable task. Given the rapidity and unpredictability with which HIV-1 genetic forms may propagate in future, a vaccine protective against all major HIV-1 circulating genetic forms is desirable, which could require multivalent formulations. Understanding the kinetics and directions of this continuing adaptation and its impact on viral fitness, immunogenicity and pathogenicity are crucial to the successful design of effective HIV vaccines. In this review, we focus on extensive diversity of HIV-1, emergence of recombinant forms and their impact on diagnosis, antiretroviral therapy, disease progression, transmission, and vaccine development. PMID- 15817946 TI - Biology of the HIV Nef protein. AB - The accessory Nef protein is expressed by all primate lentiviruses--HIV-1,HIV-2 and simian immune deficiency virus (SIV). Its expression in the early stages of the viral life cycle ensures two basic attributes of HIV infection. These are T cell activation and the establishment of a persistent state of infection. Nef has a positive effect on viral infection and replication by promoting the survival of infected cells. Its role in HIV persistence is based largely on the ability of Nef to downmodulate the surface levels of important molecules at the immune synapse. These include major histocompatibility complex-I (MHC I) and (MHC II) present on antigen-presenting cells (APCs) and target cells, and CD4 and CD28 present on helper T cells. In this review we present these biological properties of Nef from a mechanistic point of view, and relate them to the structural attributes and interactions of the Nef protein. A brief outline of the limited studies on Nef from Indian subtype C HIV-1 isolates is also presented. PMID- 15817947 TI - Molecular epidemiology of HIV. AB - The phylogenic analysis of the nucleotide sequences of the env gene has enabled classification of HIV-1 into three groups. The group M of HIV-1 infection has been classified into 9 different genetic subtypes A-K, with E and I being classified as circulating recombinants forms (CRFs). The groups O and N are less frequently encountered in human infections. Presently group M of HIV-1 globally causes 99.6 per cent of all human infections. The epidemiological trends suggest that subtype C strains would dominate the HIV pandemic in the coming years. The geographic spread of subtype C strains is also very diverse with prevalence in Africa, Latin America and Asia. Data from India show a high prevalence of subtype C. In north and western India, 78.4 and 96 per cent of HIV-1 strains respectively were shown to be subtype C. Among female sex workers in Kolkata 95 per cent of the HIV-1 strains were subtype C. The south Indian subtype data are very similar to the data from the rest of India. The HIV-2 groups (subtypes) recognized are A H. Unlike HIV-1, HIV-2 strains are predominantly found in Africa. The Indian HIV 2 strains identified till date are subtype A. This is also the predominant strain circulating in western African countries. This group (subtype) is estimated to cause 0.11 per cent of all HIV infections in humans. PMID- 15817948 TI - Low-cost monitoring of HIV infected individuals on highly active antiretroviral therapy (HAART) in developing countries. AB - The standard methods to monitor HIV infection are flow cytometry-based for CD4+ T lymphocyte count and molecular assays to quantify plasma viral load of HIV. Few laboratories in resource-limited countries can run these tests as a majority of the HIV infected individuals are poor. A number of currently available low-cost assays which require less expensive equipment and reagents, may be well-suited to such countries. These include manual and ELISA based CD4 cell assays, and ultrasensitive reverse transcriptase quantitation (Cavidi) and p24 (ELAST) assays to monitor virus load. But better internal quality assurance and quality control (QA/QC) programmes are essential. This review discusses the low-cost assays and their role in clinical monitoring of HIV infected individuals in resource-limited countries like as India. PMID- 15817949 TI - HIV/AIDS epidemic in India: risk factors, risk behaviour & strategies for prevention & control. AB - Since the first report of HIV infection in India in 1986, the virus has spread all over the country although there is geographic variation. There are estimated 5.1 million people infected with HIV with an overall estimated adult prevalence below 1 per cent. Surveys carried out in different sub-populations have yielded prevalence estimates, but data on HIV incidence are limited. Both HIV serotypes 1 and 2 exist in India and HIV-1 C is the commonest subtype reported. Sexual transmission of HIV is most predominant. Spread of HIV in intravenous drug use settings is localized mostly in the north eastern region and metropolitan cities and parent to child transmission is on the rise. Dual epidemics of HIV and tuberculosis, increase in the number of infected women, stigma and discrimination are the main concerns in the Indian HIV/AIDS scenario. There is an increasing political will and commitment for HIV prevention and control efforts in India. A multi-disciplinary approach combining targeted interventions like early identification and treatment of STDs, condom promotion, blood safety, drug de addiction programs and expanding and strengthening VCTCs and long-term strategies like awareness oriented to behavioural change especially among vulnerable populations, young people and women, steps towards improvement of literacy, status of women and overall development, reduction in poverty and development of primary prevention interventions like vaccines and microbicides will have to be considered for effective prevention and control of AIDS in India. PMID- 15817950 TI - Human immunodeficiency virus (HIV) & sexually transmitted diseases (STDs). AB - Among various factors associated with the sexual transmission of HIV, sexually transmitted diseases (STDs) seem to contribute significantly. Hence, the efforts for prevention and control of HIV have to rely largely on STD control measures. In the developing countries, both prevalence and incidence of STDs are very high. STDs impact women's health adversely for a variety of reasons such as more susceptibility than men, asymptomatic nature of infection, etc. Enormous evidence is available indicating that both ulcerative and inflammatory STDs increase the risk of HIV infection. STDs promote HIV transmission by facilitating HIV shedding in the genital tract, causing disruption of normal epithelial barrier and by deploying and activating HIV susceptible cells at the site. The effect of STD treatment intervention was studied in three controlled clinical trials conducted in Africa. Though the STD treatment intervention was shown to reduce the STD incidence in all the three studies, the decline in HIV incidence was noticed in only one of them, which, might be attributed to the differences in the study designs. To obtain reliable epidemiological data on various STDs in different communities and regions, countrywide development of adequate laboratory infrastructure for accurate diagnosis of STDs, increased outreach of awareness programmes to communities in the rural areas are some of the key issues in the fight against HIV epidemic in India. PMID- 15817951 TI - Clinical profile of HIV in India. AB - The clinical course of human immunodeficiency virus (HIV) disease and pattern of opportunistic infections varies from patient to patient and from country to country. The clinical profile of HIV disease in India includes a wide range of conditions like tuberculosis, cryptococcal meningitis, popular pruritic eruptions, and cytomegalovirus retinitis, among others. Tuberculosis is the most common opportunistic infection in Indian patients with HIV. Occurrence of various AIDS-associated illnesses determines disease progression. Mean survival time of Indian patients after diagnosis of HIV is 92 months. In this review, we discuss the clinical profile of HIV disease through an organ system-based approach. With the availability of antiretroviral therapy at lower cost, the clinical profile of HIV disease in India is now changing to include drug-related toxicities and immune reconstitution syndrome. PMID- 15817952 TI - Progress in diagnosis of opportunistic infections in HIV/AIDS. AB - The impact of HIV/AIDS is different in developed and developing countries including India. Limited access to health care facilities, lack of infrastructure for diagnostic set up and cost of anti retroviral therapy are some of the reasons. Early accurate diagnosis of opportunistic infections (OIs), the common presenting symptom of the patients, is the key for success of effective management. This review attempts an overview of few of the important OIs with which majority of Indian patients present in the clinics. Emphasis has been on conventional method of diagnostic approach, which is possible in most of the diagnostic laboratory set up in India. Awareness of the disease and maintenance of high index of clinical suspicion are required. An integrated approach to patient management with active interaction between clinicians and microbiologists would be highly beneficial. Introduction of routine in vitro antimicrobial testing system especially for Candida sp, Cryptococcus sp and Mycobacterium sp, is also important, in order to obtain a baseline data on the susceptibility pattern, which not only have therapeutic relevance, but also can predict in advance, any shift in these patterns in the Indian population. Administrative support for the skill development of personnel, facility for data preservation and telemedicine can extend the diagnostic expertise to the remote areas, without affecting the patients mobility. PMID- 15817953 TI - Diagnosis & management of leishmania/HIV co-infection. AB - Leishmaniasis, a globally prevalent parasitic disease occurs in three forms viz., visceral, cutaneous and mucocutaneous, transmitted by the bite of infected female Phlebotomus sandflies. Visceral leishmaniasis (VL) has 100 per cent fatality rate, if left untreated. India has the largest burden of this disease. HIV infection is also increasing worldwide and several reports indicate rising trend of VL/ HIV co-infection, modifying the traditional anthroponotic pattern of VL transmission. Both VL and HIV tend to lower the cell mediated immunity (CMI) resulting in poor drug response and opportunistic infections involving gastrointestinal, cutaneous, respiratory tract and central nervous system (CNS) may occur. Diagnosis of such co-infected cases is quite difficult. However, newer tests like nested PCR, rk39 immunochromatographic test etc., can be of help. Response to different antileishmanial drugs like sodium antimony gluconate (SAG), amphotericin B is far from satisfactory. However, a new oral drug miltefosine has been found to be promising. Highly active antiretroviral therapy (HAART) need to be given for management of HIV infection along with treatment of other opportunistic infections. PMID- 15817954 TI - Preventive therapy for tuberculosis in HIV infected individuals. AB - The increased risk of developing tuberculosis (TB) among those infected with HIV has prompted a need to reconsider the institution of preventive therapy/chemoprophylaxis with one or more antituberculosis drugs. Prior to the initiation of preventive therapy for tuberculosis, it is essential to rule out active TB. The target population for chemoprophylaxis among HIV seropositives includes all Mantoux (PPD) positive individuals who do not have active tuberculosis and could include all PPD negative individuals living in high prevalence region for TB. The optimal duration of preventive therapy with single drug isoniazid, daily or twice weekly, should be greater than six months to provide the maximum degree of protection against tuberculosis. The effectiveness of preventive therapy should be evaluated at regular intervals by monitoring patients for drug adherence, drug toxicity and for the development of tuberculosis. Though the impact of preventive therapy on an individual basis may be rather small, widespread implementation would have substantial impact on morbidity due to tuberculosis and some impact on mortality. Till the vast majority of HIV positive individuals in the world can access antiretroviral therapy, preventive therapy for tuberculosis should be offered at voluntary counselling and testing centres, as part of a package of care that includes prophylaxis and treatment of opportunistic infections, nutritional support and counselling. PMID- 15817955 TI - Hepatitis B & hepatitis C in HIV-infection. AB - Human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) are the three most common chronic viral infections seen in the world. All three viruses share modes of transmission and hence co-exist in the same host at significantly high rates. HIV-induced immunosuppression has deleterious effects on the natural history, pathophysiology, diagnosis, therapeutic responses to hepatitis viruses. Responses to HBV vaccination are impaired in persons with HIV infection. Co-infection with the hepatitis viruses and HIV is likely to become a major health care catastrophe in the coming years. This review discusses the current trends in the understanding of the biology of co-infection and implications for treating these viruses effectively. PMID- 15817956 TI - HIV & psychiatric disorders. AB - HIV infection and psychiatric disorders have a complex relationship. Being HIV infected could result in psychiatric disorders as a psychological consequence of the infection or because of the effect of the HIV virus on the brain. Disorders may be as varied as depression, post-traumatic stress disorders, AIDS phobias, grief and the whole gamut of cognitive disorders. In addition, several psychiatric conditions may predispose individuals to acquiring HIV infection as a consequence of their influence on behaviour. There is also strong evidence of the relationship of substance use disorders and severe mental illnesses with HIV infection. HIV related psychiatric disorders also offer a challenge to clinicians in issues of differential diagnosis and management. Majority of the work in India has focused on substance use and HIV, and to a lesser extent on the psychiatric effects of HIV infection. Given the magnitude of the problem in the country and the multiple physical and psychological stressors that persons with HIV face in India, more research is needed. PMID- 15817957 TI - Neuropathology of HIV/AIDS with an overview of the Indian scene. AB - Neurological manifestations of HIV infection and AIDS are being recognized with a frequency that parallels the increasing number of AIDS cases. Next to sub-Saharan Africa, India has the second largest burden of HIV related pathology, essentially caused by HIV-1 clade C in both the geographic locales, in contrast to USA and Europe. But the true prevalence of HIV related neuroinfections and pathology is not available due to inadequate medical facilities, social stigma and ignorance that lead to underdiagnosis. Neurotuberculosis, followed by cryptococcosis and toxoplasmosis in various combinations are the major neuropathologies reflecting the endemicity and manifesting clinically by reactivation of latent infection. Discordance in the clinical prevalence of various infections, when compared to pathological studies highlight similarities in clinical, radiological modalities of diagnosis and inherent problems in establishing definitive diagnosis. Viral infections appear to be relatively rare. Inspite of heavy burden of HIV/AIDS, HIV associated neoplasia is infrequent, including primary CNS lymphomas. HIV encephalitis and HIV associated dementia are considered infrequent, though systematic studies have just been initiated in various centres. Peripheral neuropathy characteristically manifests with vasculitic neuropathy while diffuse infiltrative lymphocytosis syndrome (DILS) involving nerves has not been reported from India. Spinal cord pathology including vacuolar myelopathy is rare, even in asymptomatic cases. Till now the AIDS cases in India were drug naive but a new cohort of cases following initiation of HAART therapy as a national policy is soon emerging, altering the biology and evolution of HIV/AIDS in India. Lacunae in the epidemiology, diagnosis and study of biology of HIV/AIDS are outlined for future research. PMID- 15817958 TI - Prevention of mother-to-child transmission of HIV--an overview. AB - With the human immunodeficiency virus (HIV) epidemic showing a shift towards women and young people, the increasing seroprevalence among women will result in an increase in the mother-to-child transmission of HIV. The vast majority of HIV positive children worldwide acquire the infection through vertical transmission. The discovery of successful interventions that interrupt this transmission has been one of the greatest successes in AIDS research. The transmission of HIV from an infected mother to her child can be reduced to less than 2 peer cent by intensive interventions in the antenatal, intranatal and postnatal periods. To achieve this low rate, primary prevention of HIV infection in parents-to-be, early identification of seropositivity in pregnant women, prevention of unwanted pregnancies, prevention of mother-to-child transmission of HIV by appropriate antiretroviral therapy, special interventions in maternal management during labour, appropriate care and follow up of the newborn, all play an important role. However, these approaches are not always possible in developing countries wherein currently 95 per cent of vertical transmission occurs. Several questions and challenges remain. These include choice, availability, affordability, duration, long-term safety of optimal antiretroviral agents to be used during pregnancy and early neonatal life and the issue of transmission via breastfeeds in situations where alternatives to breastfeeding are not available. The challenge is to find the most cost-effective and feasible intervention to achieve zero per cent transmission of HIV from an infected mother to her child. PMID- 15817959 TI - Preparing for efficacy trials of vaginal microbicides in Indian women. AB - Vaginal microbicides are topical compounds that are expected to protect women against vaginal transmission of HIV and other sexually transmitted pathogens. A large number of compounds are being evaluated as possible microbicides. Considering the spread of the HIV epidemic among women in India, clinical trials on microbicides starting from the Phase I safety and acceptability studies to Phase III efficacy trials are important. Conducting efficacy trials is a major endeavour and this review discusses challenges and issues and the preparatory steps to make such efficacy trials possible in India. PMID- 15817960 TI - Application of laboratory methods for estimation of HIV-1 incidence. AB - Estimating HIV-1 incidence (rate of new HIV-1 infections) in various populations is important to understand the current status of transmission dynamics, identify high-risk populations, monitor prevention efforts and target resources on programmes that are most effective in reducing transmissions. Recent developments in our ability to detect and distinguish recent and longterm HIV-1 infections using laboratory tests have made the measurement of HIV-1 incidence realistic and practical. These approaches most commonly rely on the properties of early HIV-1 antibodies after seroconversion as characterized by their levels, antibody avidity/affinity or antibody classes/subclasses or epitope specificity. The sensitive/less-sensitive testing strategy provided simple laboratory tools to detect recent seroconversion in a cross-sectional population. These assays are based on differences in antibody titres in recent versus long-term infections and have been used for sometime for estimating population incidence. However, recent work demonstrated limitations of this approach which included subtype-dependent performance and significant variability of "window periods", precluding its use in many areas of the world. Recently an IgG-Capture BED-EIA was developed in our laboratory which detects the increasing HIV-IgG as proportion of total IgG following seroconversion and can be used to detect recent seroconversion. The format of the assay, which includes a multi-subtype derived antigen, allows high consistency and similar "window periods" in different subtypes. This assay is now available commercially and is made specifically for population estimates of HIV-1 incidence. Due to the presence of divergent HIV-1 subtypes and the rapidly expanding HIV epidemic, it is important that the method selected is robust, performs similarly in different subtypes and is widely applicable for meaningful incidence estimates, trend analysis and comparison between populations. PMID- 15817961 TI - HIV testing technologies after two decades of evolution. AB - Over the past two decades, HIV diagnostics have been essential in detecting and monitoring infection, and continue to play a major role in saving lives throughout the world. As technology evolved, screening, confirmatory, and HIV monitoring assays have been improved and offer better alternatives to address blood screening, surveillance, diagnosis, and patient management. Molecular methods are critical in detecting early infection and for managing patients on anti retroviral therapy whose viral infection may become resistant to therapy. In addition, modifications to conventional methods have introduced new assays, such as sensitive/less sensitive (detuned) assays that can estimate when someone was infected, thereby providing a useful tool for epidemiologic incidence estimates and enrollment into specific intervention programmes for recently infected persons. Many of the newly evolving technologies are essential for use in resource-limited countries because they can address cost issues, limited infrastructure, and a lack of formally trained personnel. Newer rapid HIV kits can be stored in a wide range of temperatures (2-30 degrees C) to address cold chain issues, can use easily-collected fingerstick blood and oral fluids, and have one-step procedures that are relatively foolproof. Manual CD4 lymphocyte count assays that require only a light microscope and haemacytometer and more simple assays to estimate viral load are appropriate for developing countries where sophisticated instrumentation cannot be supported. Technologic advances with HIV diagnostics continue to address outstanding and new issues associated with diagnosis and the monitoring of infection by providing more simplified, cost effective, and accurate testing throughout the world. PMID- 15817962 TI - CD4+ T cell count as a tool to monitor HIV progression & anti-retroviral therapy. AB - The CD4+ T lymphocytes are the crucial cells in the orchestral events in forming immune response to the foreign antigen and it is also the primary target cells for human immunodeficiency virus (HIV). The progressive loss of these cells eventually results in the loss of an ability to mount desirable immune response to any pathogen and death of the patients in the terminal stage of HIV infection, i.e., acquired immune deficiency syndrome (AIDS). The longitudinal monitoring of CD4+ counts is used as a monitoring tool for disease progression and effectiveness of antiretroviral treatment. Various methods are being used for determination of absolute CD4+ T lymphocytes from the peripheral blood. To date, the flow cytometry is considered as the gold standard. Different modifications have been tried in the conventional flow cytometry to increase accuracy and cost effectivity especially for adapting in resource-poor settings. Principles of the conventional and modified methodologies are discussed. The non-flow cytometric methodologies are also there that might be available soon widely. The choice of the methodology should depend upon the purpose of the assay, the age group of the patients, sample turnover and available resources. Importance of maintaining both internal and external quality control systems in every laboratory performing CD4 count estimation are discussed. PMID- 15817963 TI - HIV-TB co-infection: epidemiology, diagnosis & management. AB - HIV/AIDS pandemic has caused a resurgence of TB, resulting in increased morbidity and mortality worldwide. HIV and Mycobacterium tuberculosis have a synergistic interaction; each accentuates progression of the other. Clinical presentation of TB in early HIV infection resembles that observed in immunocompetent persons. In late HIV infection, however, TB is often atypical in presentation, frequently causing extrapulmonary disease. These factors coupled with low sputum smear positivity, often result in a delayed diagnosis. HIV-infected patients respond well to the standard 6-month antituberculosis treatment regimens, although mortality is high. Antituberculosis treatment is complicated by frequent drug interactions with highly active antiretroviral therapy (HAART) and adverse drug reactions are more common among HIV-infected patients. Guidelines for the management of patients co-infected with HIV and TB are still evolving. Timely institution of antituberculosis treatment using the directly observed treatment, short-course (DOTS) strategy and HAART markedly improves the outcome of HIV infected patients with TB. PMID- 15817964 TI - AIDS vaccine development: perspectives, challenges & hopes. AB - The worldwide quest for an AIDS vaccine represents an unprecedented scientific and human challenge for the 21st century. Preventive vaccines represent our only long-term hope to stop the epidemic. AIDS vaccines must be seen as the ultimate prevention tool that will complement the existing prevention strategies in place. The acceleration of vaccine development through the parallel exploration of several scientific approaches and implementation of clinical trials are the best and probably only way to reach this goal, and the best vaccines have moved into phase II and efficacy trials. Ideally an AIDS vaccine should induce both neutralizing antibodies against HIV-1 primary isolates and cell-mediated responses. AIDS vaccines could prevent either HIV infection or progression to disease and decrease transmission by reducing the HIV viral load. Most of the vaccine approaches developed so far aim at inducing cell-mediated immune responses. New vector-based vaccines include modified vaccinia Ankara, adeno associated virus, adenovirus and alpha viruses. Considerable efforts are on to develop vaccines that would induce neutralizing antibodies. All vaccines tested so far in humans have proven to be safe. This long-term endeavour requires strong and renewed political leadership and commitment, flexibility of processes, medical and scientific dedication and collaboration on a mission mode along with community participation for immediate action. Recent developments in India highlight clearly the commitment of the Government of India and the scientific community to a long-term global effort to develop an AIDS vaccine. PMID- 15817965 TI - HIV/AIDS epidemic in India: an economic perspective. AB - Whilst the international community's focus has been on the region most devastated by HIV/ AIDS, namely sub-Saharan Africa, India now appears on the brink of a significant AIDS epidemic. In thinking about the implications of HIV/AIDS, considerable attention was initially drawn to its clinical aspects. More recently, other dimensions of HIV, including economic, have been explored. The primary objective of this review is to elaborate on the major elements of the national and international economic research to data on HIV/AIDS, and to infer lessons from it, for India. It also examines the evidence on the aggregate and household-level economic impacts of HIV, the economic roots that drive its transmission and the methods economists use to assess the efficacy of alternative interventions to address HIV and AIDS. Available evidence suggests that whereas aggregate impacts may be limited, the adverse household-level economic implications of AIDS may be serious; public resources that are available for health are also likely to be put under strain. Paucity of economic research on HIV and AIDS relating to India is highlighted. PMID- 15817967 TI - Web watch: some Web-based resources for HIV/AIDS. PMID- 15817966 TI - Ethical issues in HIV/AIDS research. AB - Globally the HIV/AIDS epidemic has presented unique health challenges to populations, including a host of ethical and moral issues related to human life and dignity. The disease has most affected the vulnerable groups of people in the world often leading to stigma and discrimination. Currently the critical areas of concern include access to treatment and developing newer, more effective therapeutic and prevention methods while taking care of ethical values in health care and research. The ethical issues mainly revolve around the standard of care, informed consent across cultures, privacy and confidentiality, stigma and discrimination, protection of vulnerable groups, community consultation, ethical review mechanisms, international collaboration, epidemiological studies, clinical trials and also sociobehavioural studies on HIV/AIDS. In addition major concerns are raised regarding HIV/AIDS vaccine trials, microbicidal trials as well as prevention of parent-to-child transmission (PPTCT) trials especially in resource poor countries. For international collaborative research appropriate standard of care, community benefits, and host country needs must be kept in mind. Improving our understanding of the various ethical and societal concerns related to HIV/AIDS treatment and research would help in development of appropriate policies for disease control and prevention. PMID- 15817968 TI - Protecting IMS Act: balancing trade interest and child health. PMID- 15817969 TI - Late Hony. Surg. Cmde. Dr. Shantilal C. Sheth Oration. Presentation during PEDICON 2005 Kolkata, January 7th, 2005. Changing trends--a challenge to the "already trained". PMID- 15817970 TI - Randomized evaluation of fluid resuscitation with crystalloid (saline) and colloid (polymer from degraded gelatin in saline) in pediatric septic shock. AB - OBJECTIVE: To compare the efficacy of crystalloid (Normal saline) and colloid (polymer from degraded Gelatin in saline Haemaccel) intravenous fluid in restoration of circulating volume in children with septic shock. DESIGN: Prospective, randomized, open-label trial. SETTING: Pediatric Emergency and Intensive Care Unit of a tertiary care referral and teaching hospital. SUBJECTS AND INCLUSION CRITERIA: Sixty patients, between 1 month to 12 years of age, with septic shock, without clinical evidence of organ failure at admission or underlying immunodeficiency. INTERVENTION: Resuscitation with normal saline or polymer from degraded gelatin (Haemaccel) in the boluses rate of 20 mL/kg till hemodynamic stabilization or if central venous pressure (CVP) exceeded 10 mmHg (fluid requirement beyond 40 mL/kg guided by BP and CVP). METHODS: Hemodynamic parameters (heart rate, capillary filling time, pulse volume, and blood pressure) were recorded before and during resuscitation, and then 2 hourly for 12 hours. Central venous pressure line was placed within first hour, soon after starting fluids. Estimation of plasma volume and body water was done at the end of first hour of fluid resuscitation. OUTCOME MEASURES: Hemodynamic stabilization (heart rate, capillary refill time, systolic BP in normal range), plasma volume at the end of fluid resuscitation and incidence of organ dysfunction. RESULTS: 31 patients were randomized to normal saline and 29 to gelatin polymer. Both the groups were similar with respect to age, gender, primary diagnosis, initial hemodynamic parameters and PRISM score. Pneumonia (n = 22; 36%), gut-associated sepsis (n = 13), and dengue hemorrhagic fever (n = 11) were the common primary diagnosis. Initial hemodynamic stabilization was achieved in all. The mean (SD plasma volume (saline--53.4 (2.0 mL/kg, gelatin polymer--53.2 (1.9 mL/kg), extracellular fluid volume, total body water and interstitial fluid volume at the end of first hour of resuscitation were similar. The requirement of inotropes, incidence of organ dysfunction and case fatality rate (Saline--29%, gelatin polymer--31%), were similar in two groups. CONCLUSION: Both normal saline and gelatin polymer solution were equally effective as resuscitation fluid with respect to restoration of plasma volume and hemodynamic stability. Normal saline upto 110 mL/kg, and gelatin polymer solution upto 70 mL/kg may be required in first hour for successful fluid resuscitation of septic shock in children. PMID- 15817971 TI - All slums are not equal: child health conditions among the urban poor. AB - Increasing urbanization has resulted in a faster growth of slum population. Various agencies, especially those in developing countries are finding it difficult to respond to this situation effectively. Disparities among slums exist owing to various factors. This has led to varying degrees of health burden on the slum children. Child health conditions in slums with inadequate services are worse in comparison to relatively better served slums. Identification, mapping and assessment of all slums is important for locating the hitherto missed out slums and focusing on the neediest slums. In view of the differential vulnerabilities across slums, an urban child health program should build context appropriate and community-need-responsive approaches to improve children's health in the slums. PMID- 15817972 TI - Role of shunt surgery in pediatric tubercular meningitis with hydrocephalus. AB - This study was designed to evaluate the indications for ventriculoperitoneal shunting in cases of children with tubercular meningitis, presenting with hydrocephalus. Thirty seven children (less than 18 years of age) of tubercular meningitis with hydrocephalus (TBMH) who underwent ventriculoperitoneal shunting over a three year period (1999 to 2001) were included in the study. Sixteen (43%) children were Palur stage II, 15 (40%) stage III, and 6 (16%) stage IV. Fifteen (40%) children had received antitubercular therapy for less than 4 months and 17 (46%) received therapy for more than 4 months prior to presentation. Five (14%) children had not previously received antitubercular therapy. Shunt related complications occurred in 11 (30%) children and 3 children had undergone revision of the shunt multiple times. Good outcome was seen in 16 (43%) children. Thirteen (35%) had moderate disability and 6 (16%) had severe disability at 3 months of follow up. 62% (n =10) children in grade II had a good outcome compared to 40% (n = 6) in grade III. All six children in grade IV had a poor outcome. 2 children, both having multiple infarcts, died and the remaining 4 were left with severe disability. We recommend shunt placement in all children of grade II and III TBMH as this policy has yielded the best results. For grade IV children external ventricular drainage, followed by shunting if improvement occurs remains the most cost-effective procedure. PMID- 15817973 TI - Correlation of serum parathormone level with biochemical parameters in chronic renal failure. AB - A prospective study was carried out to assay the level of serum intact parathormone and its correlation with biochemical parameters in patients with chronic renal failure (CRF). The study included 64 children (44 with CRF, and 20 age and sex matched controls). Serum intact parathormone (iPTH), serum creatinine, urea, calcium, inorganic phosphate and alkaline phosphatase were estimated. Creatinine clearance (Ccr) was estimated by Schwartz formula. Patients with CRF were divided into four groups based on their Ccr (mild CRF with mean Ccr 59.17 +/- 1:18.53 mL/min/1.73 m2 (n = 6) moderate CRF with mean Ccr 34.98 +/- 7.75 mL/min/1.73 m2 (n = 7); severe CRF with mean Ccr 17.71 +/- 5.40 mL/min/1.73 m2 (n = 15); and end-stage renal disease with mean Ccr 6.46 +/- 1.71 mL/min/1.73 m2 (n = 16). Mean serum iPTH levels were 93.00 +/- 46.62 pg/mL in CRF and 16.52 +/- 9.35 pg/mL in controls. Groupwise mean serum (iPTH) levels were 48.50 +/- 4.76, 67.29 +/- 7.91, 82.42 +/- 9.67 and 130.66 +/- 58.74 pg/mL in mild, moderate, severe CRF and endstage renal failure respectively. Mean serum iPTH level of CRF (93.00 +/- 46.42 pg/mL) negatively correlated with mean Ccr (22.02 +/- 18.53 mL/min/l.73 m2) (P < 0.001) and mean serum calcium (7.30 +/- 1.02 mg/dL) (P < 0.001) and positively correlated with mean inorganic phosphate (5.76 +/- 1.1 mg/dL) (P < 0.05) and mean alkaline phosphatase (355.14 +/- 185.53 UL) (P < 0.001). We conclude that increased iPTH level occur even early in the course of CRF and progressive hypocalcemia and hyperphosphatemia are the initiating factors for the development of hyperparathyroidism. PMID- 15817974 TI - Transcutaneous absorption of oil in preterm babies--a pilot study. AB - This study was conducted to determine transcutaneous absorption of oil in preterm neonates. A mixture of coconut oil and Meadowfoam oil which contains unique fatty acids, which acted as marker fatty acids was applied to the skin of babies. One mL blood was collected before and one hour after post oil application. Both pre and post oil application serum samples were hydrolysed and derivatised with 2 phenyl hydrazine hydrochloride in order to detect fatty acids by HPLC analysis on C-8 column. None of the pre oil application serum sample showed the presence of the marker fatty acids. The post oil application serum sample of all the 12 babies showed the presence of marker fatty acids of Meadowfoam oil which indicates transcutaneous absorption of oil in preterm babies. PMID- 15817975 TI - Computerized tomography detects pulmonary lesions in children with normal radiographs diagnosed to have tuberculosis. AB - This report is based on observations during the conduct of a larger study to develop diagnostic criteria for childhood tuberculosis (TB). Of 201 children confirmed to have pulmonary or lymph node TB, 84 had normal chest radiographs. Computerized tomography (CT) of the chest was performed in nine of them, seven of whom had normal chest radiographs while two had visible calcification. Eight of the nine children had definitive lesions detected by computerized tomography of the chest. While five children had primarily hilar lymph node enlargement, three had pulmonary parenchymal lesions. The use of more sensitive diagnostic tests like computed tomography helps to detect tuberculosis lesions not otherwise visualized on chest radiographs. This report highlights the difficulty in excluding active tuberculosis in children. More studies are required on the role of CT scans in the diagnosis of tuberculosis in children. PMID- 15817976 TI - Body mass index in Bengali adolescents. AB - We describe a mixed longitudinal analysis of body mass index (BMI) in a group of Bengali adolescents (age 11-17 years) from a middle income family background and compare this against existing national and international data. Healthy school children, comprising of 416 boys and 343 girls were consented for annual repeat measurements of weight, height and pubertal staging between the years 1998 and 2001. The LMS method was used to construct smoothed BMI mean and standard deviation (SD) curves. Bengali adolescents have lower BMI than affluent Indian children and are -1 to -2 SD below US children. BMI increases in adolescence (boys: r = 0.49, p < 0.001, girls: r = 0.54, P < 0.001) with age but SMR does not have an independent effect on BMI. PMID- 15817977 TI - Ten commandments for Child Friendly School Initiative. PMID- 15817978 TI - Sternal malformation: vascular dysplasia complex. PMID- 15817979 TI - Sclerocornea. PMID- 15817980 TI - Van der Woude syndrome. PMID- 15817981 TI - Neutropenic enteropathy. AB - Neutropenic enteropathy (NE) is used to describe the inflammation of the bowel in neutropenic patients under aggressive chemotherapy, mainly for lymphoproliferative and hematologic malignancies. Surgical intervention may be required in patients with the advent of the disease. We report our experience in 7 children with NE who had to be treated surgically. Absolute neutrophil counts were less than 1000/mm3 in all, with positive blood cultures in five patients. Four patients recovered with rapid resolution of neutropenia, while three patients died with persistent neutropenia. PMID- 15817982 TI - Hereditary sensory autonomic neuropathy Type IV. AB - Hereditary sensory autonomic neuropathy Type IV is an autosomal recessive disorder due to lack of maturation of small myelinated and unmyelinated fibers of peripheral nerves, which convey sensation of pain and temperature, therefore, resulting in self mutilation. There is anhidrosis due to lack of innervation of normal sweat glands resulting in recurrent episodes of hyperpyrexia. The clinical presentation of two children with this rare disease is described. PMID- 15817983 TI - Fanconi's anemia in newborn. AB - Fanconi's anemia (FA) is a paradigm for congenital anomalies, aplastic anemia and predisposition to malignancies. Identification of the disease at birth is based on characteristic physical malformations, as hematologic manifestations at birth are extremely rare. We report a case of FA in a newborn who presented with anophthalmia, unilateral radial ray defect, hemivertebrae and thrombocytopenia. PMID- 15817984 TI - Massive powder aspiration in a toddler. AB - We report a case of aspiration of calcium carbonate powder by a toddler. Bronchoscopic removal of aspirated contents resulted in favourable outcome. PMID- 15817985 TI - Pyridoxine dependent and pyridoxine responsive seizures. PMID- 15817986 TI - Transverse testicular ectopia. PMID- 15817987 TI - Neonatal hypocalcemia due to asymptomatic maternal primary hyperparathyroidism. PMID- 15817988 TI - Nephrocalcinosis in a child with nephrotic syndrome. PMID- 15817989 TI - Tongue biting and epilepsy. PMID- 15817990 TI - Double fistulae in esophageal atresia. PMID- 15817992 TI - DOTS in pediatric patients. PMID- 15817993 TI - Glutathione peroxidase mimics as novel antioxidants from vegetables. AB - Vegetables are generally recognized as rich sources of dietary antioxidants for inhibiting lipid peroxidation. Here we investigated lipid hydroperoxide (LOOH) reducing activity of several vegetables to estimate their role on the prevention of lipid peroxidation in food and the digestive tract. By using HPLC analysis, we screened vegetables possessing the ability to convert 13 hydroperoxyoctadecadienoic acid (13-HPODE) to its reduced derivative, 13 hydroxyoctadecadienoic acid (13-HODE). Welsh onion (Allium fistulosum L.) was found to be highly active in the reduction of 13-HPODE among tested vegetables. There was no relationship between 13-HPODE reducing activity and GSH peroxidase (GPX) activity in the tested vegetables. 13-HPODE-reducing activity of welsh onion was enhanced by the addition of sulfhydryl compounds including glutathione (GSH). Neither GPX inhibitor nor heat treatment suppressed 13-HPODE-reducing activity effectively. These results suggest that welsh onion and other vegetables contain GPX mimics responsible for the reduction of LOOH. GPX mimics may be helpful in the attenuation of harmful effect of LOOH from food. PMID- 15817994 TI - Treatment of massive hypertriglyceridemia resistant to PUFA and fibrates: a possible role for the coenzyme Q10? AB - OBJECTIVE: To describe the effect of CoQ10 (added to either a fibrate, or PUFA or association of both) in patients affected by massive hypertriglyceridemia (MHTG) resistant to fibrates and PUFA. DESIGN: Open, sequential, comparative intervention study. SETTING: Specialised centres for dyslipidemia management. SUBJECTS: 15 subjects (mean age: 45.1 +/- 12.5 years) affected by MHTG and hyporesponsive to either fibrates, or PUFA, or fibrates-PUFA association, and 15 age-matched subjects regularly responders to PUFA and fenofibrate treatment. INTERVENTIONS: Treatment for periods of 6 weeks each with the following consecutive treatments: CoQ10 150 mg/day, PUFA 3000 mg/day, fenofibrate 200 mg/day, PUFA 3000 mg/day + fenofibrate 200 mg/day, PUFA 3000 mg/day + CoQ10 150 mg/day, fenofibrate 200 mg/day + CoQ10 150 mg/day, and finally, fenofibrate 200 mg/day + PUFA 3000 mg/day + CoQ10 150 mg/day. RESULTS: CoQ10 supplementation improved, in the control group, systolic and diastolic blood pressure, creatinine and Lp(a) plasma levels, both during fenofibrate and/or PUFA treatment. In MHTG group, CoQ10 supplementation significantly improved TG, TC, Lp(a), uric acid and blood pressure during fenofibrate treatment, but only Lp(a) and blood pressure during PUFA treatment. Fenofibrate appeared to have better effect on hsCRP and gamma-GT plasma levels than PUFA. No significant change was observed in any group and under any treatment in regards to homocysteinemia, PAI-1, or t-PA. CONCLUSION: Even though the mechanism of action through which the effects were obtained is yet to be elucidated, adding CoQ10 to fenofibrate could improve the drug's efficacy in MHTG patients not responding to fenofibrate alone. PMID- 15817995 TI - Minoxidil, a K(ATP) channel opener, accelerates DNA synthesis following partial hepatectomy in rats. AB - A large number of studies have reported the action of K(ATP) channel openers in accelerating the proliferation of hepatocytes and many other cell types in vitro. Few studies, however, have examined the proliferative effect of K(ATP) channel openers in vivo. The aim of this study was to determine whether the K(ATP) channel opener minoxidil accelerates liver regeneration after partial hepatectomy (PH) in vivo. Male Wistar rats underwent a 70% partial hepatectomy (PH) after receiving a subcutaneous injection of minoxidil (0.01 mg/kg or 0.03 mg/kg). Some of the rats were intravenously treated with 5-hydroxydecanoic acid (5-HD, 10 mg/kg) just before the minoxidil injection. Seventy-two hours after PH, DNA synthesis was immunohistochemically assessed by bromodeoxyuridine (BrdU) incorporation into the nuclei. Minoxidil induced significant and dose-dependent increase in the BrdU labeling index after PH, and 5-HD reversed this minoxidil induced change. Minoxidil did not significantly affect the changes in liver weight and liver function after PH. The hepatic levels of prealbumin decreased by about 60% after PH and minoxidil inhibited the decrease. In conclusion, the K(ATP) channel opener minoxidil enhanced DNA synthesis after PH without affecting the liver function. PMID- 15817996 TI - EGCG, a major component of green tea, inhibits VEGF production by swine granulosa cells. AB - Vascular Endothelial Growth Factor (VEGF) plays a pivotal role in the physiological ovarian angiogenic process: its production appears to be stimulated by the hypoxic environment which takes place during follicle development. Recently, epigallocatechin-3-gallate (EGCG) from green tea has been used in livestock nutrition as an alternative to antibiotics. However, despite many potential benefits of EGCG consumption, it is also important to get an insight on the possible reproductive-related consequences of feeding supplementation: in fact this substance has been found to inhibit angiogenesis, a process fundamental for follicle development. Therefore, we evaluated the effect of EGCG (5 and 50 microg/ml) on the production of the main angiogenetic factor, VEGF, by swine granulosa cells cultured in normoxia (19% O2), partial (5% O2) or total hypoxia (1% O2). In addition, we studied the effect of the catechin on cell proliferation. Our data demonstrate that both partial and total hypoxia stimulated VEGF production. EGCG reduced VEGF production independently of the O2 condition: 50 microM was the most effective doses. Granulosa cell proliferation was inhibited by EGCG even if only by the highest concentration. This effect might possibly be due to the decrease induced in VEGF production. Therefore feeding supplementation with EGCG should be carefully considered. PMID- 15817997 TI - Lipid peroxidation in plasma of rats treated with ferric-nitrilotriacetate, in relation to kidney and liver modifications. AB - Intraperitoneal injection of the iron chelate ferric-nitrilotriacetate (Fe-NTA) induces in rodents renal and hepatic suffering, associated with oxidative damage. We investigated the oxidation pattern in plasma of treated rats in relation to liver and kidney, monitoring the variation of the lipid components more susceptible to oxidation, unsaturated fatty acids (UFA) and alpha-tocopherol, as biomarkers of the oxidative damage. A sublethal dose of Fe-NTA induced a strong and extremely significant decrease of UFA levels at 1 h after injection in the plasma compartment and at 3 h in the kidney, with reductions up to 40-50% of the control values, together with an increase of conjugated dienes fatty acids hydroperoxides and a consumption of alpha-tocopherol. The same modifications were observed in the liver, but to a lesser extent. Histological observation proved that biochemical changes in the lipid fraction were a direct consequence of an ongoing membrane lipid peroxidation process. Our data show that oxidative damage to the lipid fraction is initially evident in the plasma compartment, where Fe NTA toxicity is assumed to be caused by the elevation of serum free iron concentration, and proceeds with different speed and severity in the kidney and liver. PMID- 15817998 TI - Increase in selenium requirements with physical activity loads in well-trained athletes is not linear. AB - Selenium requirements in athletes are supposed to be increased with energy expenditure (EE) to preserve selenium status and an optimal antioxidant balance. The question of whether selenium intakes are related to EE and whether plasma selenium status induces up-regulation in erythrocyte endogenous antioxidant defense and decreases plasma oxidative damage markers in athletes was addressed. 118 well-trained athletes completed 7 d food and activities records in a cross sectional study. Blood was sampled on day 8. Among the athletes, 23% of the males and 66% of the females had selenium intakes below two-third of the French RDA. Plasma selenium concentrations in most of less trained athletes were lower than the postulated concentration to be required to maximize erythrocyte GSH-Px activity. Athletes with the highest daily EE had the highest selenium intakes, percentage of vegetal protein intakes and plasma selenium concentrations. Only 2.6% of the athletes exhibited low plasma selenium concentrations (< 0.75 micromol/l). The relation between plasma selenium and EE was polynomial (r = 0.50; P < 0.005). Erythrocyte GSH-Px activity in athletes was not linked to selenium status. Selenium requirements are increased in athletes without being linearly related to EE. PMID- 15817999 TI - [Complete denture occlusion considered from occlusal contacts during mastication]. AB - Few reports describe the functional superiority of full balanced occlusion to that of lingualized occlusion. Recently, a bilateral balanced scheme has been much more generally applied than a unilateral balanced scheme in lingualized occlusion as well as in full balanced occlusion. Occlusal contacts on the non chewing side occur earlier than on the chewing side;in the order of second molar, first molar, and then premolars. The contact on the balancing side contributes to the prevention of denture dislocation and guidance from eccentric positions to the centric occlusal position during mastication. Therefore, bilateral balanced occlusion was shown to be effective for denture stability during mastication with complete dentures. Today's typical edentulous patients have greater mean age than in the past, and thus are seen with severe alveolar bone resorption, thin mucosa and sometimes abnormalities of the maxillomandibular relation. Their occlusal positions are often unstable and changeable after insertion of complete dentures. Because lingualized occlusion allows for easier accommodation and correction, lingualized occlusion is more suitable for such cases than full balanced occlusion, which requires a strict occlusal relationship. In light of the standard Japanese diet, there is some doubt about why lingualized occlusion might be suitable for Japanese edentulous patients in terms of the sense of mastication. A method of evaluating the sense of mastication needs to be established. PMID- 15818000 TI - [Selection of occlusal scheme on the basis of pressure distribution on supporting structures under complete dentures]. AB - In case of making complete dentures, we have to consider not only denture stability but also the restoration of aesthetics and function such as mastication and speech. However these are contradictory theoretically from the point of view of denture stability, and it is very difficult to satisfy both requirements in the case of a patient who has poor upper and lower alveolar ridges. We investigated the effect of artificial posterior teeth form and occlusal scheme on the distribution of pressure on supporting structures under complete dentures during mastication with upper and lower edentulous simulators. In this report, a guideline for the selection of occlusal scheme for complete dentures, based on our previous investigations, is described. The occlusal scheme remarkably affected the distribution of pressure under simulated complete dentures, as shown by comparing the distribution of pressure using two different occlusal schemes:fully balanced occlusion and lingualized occlusion. However other factors such as posterior teeth form and position affect the distribution of pressure as well, and are related to each other. Therefore, not only occlusal scheme but also posterior artificial teeth form has to be considered, and the form of posterior teeth should be carefully and comprehensively decided when making complete dentures. PMID- 15818001 TI - [Selection of occlusal scheme on masticatory function in denture wearers]. AB - The characteristics of full balanced occlusion (FBO) and lingualized occlusion (LO), which are occlusions that make up removable dentures, were investigated from the standpoint of masticatory function, and an attempt was made to clarify criteria for selecting and configuring occlusions to suit individual cases. Since there have been few studies that have quantitatively compared FBO and LO from the aspect of masticatory function, we decided to organize and present the characteristics of both in this paper based on our findings from studies carried out in our department. We found that LO offered a higher ability of food crushing, showed higher masticatory performance in the case of hard foods, displayed faster as well as smoother masticatory movement, and showed chewing patterns that were closer to the chopper type compared with FBO. Moreover, subjective evaluation by subjects indicated that "it was easier to eat and the food tasted better" with the LO compared with the FBO in the case of general foods. Furthermore, a comparison of anatomical form and bladed form of upper lingual cusps in the case of LO showed no difference between the anatomical form and the bladed form in terms of masticatory performance in the case of any of the test foods, but the bladed form offered a higher ability of food crushing, displayed faster and smoother masticatory movement, and showed chewing patterns that were closer to the chopper type. In addition, subjective evaluation by subjects showed that it was easier for them to eat raw carrots and pickled radish, which are hard foods generally thought to be difficult to eat with dentures, with the bladed form compared with the anatomical form. PMID- 15818002 TI - [Criteria for applying lingualized occlusion by the condition of residual alveolar ridge]. AB - Lingualized occlusion is applied to patients with alveolar ridge resorption, and it is considered that this occlusion is functionally useful and should be applied to cases with alveolar ridge resorption. However, selection criteria supported by evidence for what conditions of the alveolar ridge lingualized occlusion should be selected do not exist. For that reason, we conducted an investigation on choosing lingualized occlusion. As a result, no differences were noted in subjects with a good alveolar ridge condition while a sufficient lingualized occlusion effect was obtained in subjects with a poor alveolar ridge condition, suggesting that efficient mastication could be attained. The following indexes were examined as the criteria for applying lingualized occlusion: The ratio of the residual alveolar ridge should be less than approximately 0.5, in which the distance between the inferior border of the mental foramen and the superior border of the mandibular bone should be less than half the distance between the inferior border of the mental foramen and the inferior border of the mandibular bone. The supporting area of the denture base should be less than approximately 2,000 mm(2). Though the effect can be expected when applying lingualized occlusion under these criteria, they are one standard, and cannot be applied in all cases. In clinical cases, it is important to grasp the local and the whole body conditions of the patient in order to choose denture occlusion. PMID- 15818003 TI - [Chromatic study of all-ceramic crown--IPS Empress: difference of color by manufacturing technique and cements]. AB - PURPOSE: In recent years,aesthetic appearance and function are called for and all ceramic crowns are spreading. By choosing an all-ceramic crown the problem of metal ceramics is avoided. There are difficulties of color tone reproducibility of cervical margin and darkness of gingival margin. We examined IPS Empress also in various all-ceramic crowns. IPS Empress has high permeability a ceramic ingot of various color tones and excellent color tone reproducibility of natural teeth. Generally a layering technique is used for an anterior tooth and the staining technique is used for a molar. However the details are unknown We examined how differences of manufacturing method and cement affect the color tone of all ceramics clinically. METHODS: Two kinds of Empress crown were fabricated for a 27 year-old woman's upper left-side central incisors:the staining technique of IPS Empress and the layering technique of IPS Empress II. Various try-in pastes(transparent opaque white white and yellow) of VariolinkII of the IPS Empress System were used for cementing. Color was measured using a spectrophotometer CMS 35FS. The L*a*b* color system was used for showing a color. The right-side central incisors on the opposite side of the same name teeth were used for comparison. RESULTS: We analyzed the color difference (DeltaE* ab)with a natural tooth. Consequently when it had no cement of staining technique and was tranceparent small values were obtained. CONCLUSIONS: It is considered that the color tone can be adjusted by color cement. It is effective to use the staining technique for an anterior tooth crown depending on the case. The crown manufactured using the layering technique is not easily influenced by cement. The crown manufactured by the staining technique tends to be influenced by cement. PMID- 15818004 TI - [Human papillomavirus infection in the oral cavity of denture wearers]. AB - PURPOSE: The purpose of this study was to determine the frequency of human papillomavirus(HPV)infections in the oral cavity of middle-aged and elderly dental patients with dentures. METHODS: In this study, 47 patients (20 men and 27 women), aged from 50 to 78 years, were randomly selected from the patients in the Department of Prosthodontics, Aichi-Gakuin University Affiliated Dental Hospital. Oral squamous cells were collected from swabs of the buccal mucosa. For this procedure, informed consent was obtained. Extracted DNA was evaluated for HPV infections by PCR methods, using consensus and specific primers, and direct DNA sequencing analysis. RESULTS: Twenty-four of 47 specimens (51.1%) were positive for HPV DNA. A statistically significant association was not found in the HPV positivity between men and women. The high rate of infection was recognized from 60 or more years old. A statistically significant association was found in the HPV positivity between non-denture wearers and denture wearers. Frequent HPV types in the specimens of all were HPV11, 4 and 16. Frequent HPV types in the specimens of non-denture wearers and denture wearers were HPV4, 11 and HPV11, 16 and 4, respectively. CONCLUSIONS: The results of the present investigation indicate that HPV is present in the oral cavity, especially those of denture wearers, of middle-aged and elderly patients. It is suggested, therefore, that the oral cavity of middle-aged and elderly patients with dentures is a reservoir of HPVs where later HPV-associated diseases, such as oral cancer and other oral lesions, may develop. PMID- 15818005 TI - [Differences in whole salivary total protein concentration and protein fractions among the groups of dentulous subjects, edentulous subjects and periodontitis patients]. AB - PURPOSE: To apply salivary proteins to the diagnosis of periodontal disease in partial denture supporting teeth, total protein concentrations and protein fractions in whole saliva were compared among dentulous subjects, edentulous subjects and periodontitis patients. MATERIALS AND METHODS: Eighty-five subjects/patients in total were studied, who consisted of 52 dentulous subjects with diagnosed normal periodontal tissue, 18 edentulous subjects using complete dentures and 15 patients with diagnosed periodontal disease. Total protein concentration in whole saliva was measured using dot blotting-silver staining, and protein fractions were analyzed using cellulose acetate membrane electrophoresis and silver staining. Based on the results obtained and some oral cavity examination items, the correlation between the periodontal pocket depth and the number of residual teeth was studied. RESULTS: Total protein concentration was highest in the group of periodontal disease patients and significantly different from that in the group of dentulous subjects (p<0.01). The percentage area and concentrations of albumin fraction were largest in periodontal patients, followed by dentulous subjects and edentulous subjects in this order. A significant difference (p<0.01) was observed in albumin concentration between periodontal patients and edentulous or dentulous subjects. Concentrations of immunoglobulin A and Gamma-globulin were significantly higher in periodontal patients than in dentulous subjects (p<0.01). In addition, among oral cavity examination items, the total periodontal pocket depth, the number of residual teeth and the number of teeth with periodontal pocket depth of over 4 mm were significantly correlated with the percentage area of albumin fraction or albumin concentration. CONCLUSIONS: Albumin concentration in whole saliva was suggested to be most effective for diagnosing periodontal disease in supporting teeth. PMID- 15818006 TI - [The effectiveness of prosthodontic mandibular advancement for obstructive sleep apnea syndrome]. AB - PURPOSE: This study investigated the hypothesis of a direct association between the shift of incisal point and the diameter of the oropharynx space, and nocturnal oximetry with and without prosthodontic mandibular advancement (PMA) which is one type of oral appliance used to treat obstructive sleep apnea syndrome (OSAS), and to indicate the effects of PMA. METHODS: A total of 19 patients with OSAS were subjected to finger pulse oximetry recording and lateral cephalometric radiography: with and without PMA. The distance of the oropharynx space (PNS-AS, SPAS, MAS, and IAS) and the shift of incisal point with PMA were measured. The oxygen desaturation index (ODI 4%) and the cumulative percentages of time spent at saturations below 90% (CT(90)) during sleeping were calculated. Spearman's rank correlation test was used to measure the relationship between the increase of the velopharyngeal space and the distance of the mandible. The ODI 4%, CT(90) and distance of the velopharyngeal space were compared for all subjects with and without PMA by Wilcoxon's linking test. RESULTS: There was a strong relationship between the increase of the lower part (IAS) and the distance of anterior shift of incisal point (r=0.61, p=0.01). ODI 4% and CT(90) significantly decreased (p<0.01) and the three liners of the oropharynx(SPAS, MAS, IAS) significantly increased with PMA (p<0.01). CONCLUSIONS: It was shown that prosthodontic mandibular advancement could reduce the ODI 4% and CT(90) value in patients with OSAS, by enlarging the oropharynx space. PMID- 15818007 TI - [Prediction of mandibular movement by observation of facial skin markers]. AB - PURPOSE: Various methods have been used for observation of mandibular movement. However, many methods disturb physiological movement as the subject must wear a sensor. Therefore, to predict mandibular movement from the motion of facial skin, the correlation between the movements of mandibular incisor-tooth point and facial skin markers was evaluated by multiple regression analysis. METHODS: Nine retro-reflective markers were placed on the lower facial skin of 8 dentulous subjects and three markers on the wire frame attached to the mandibular tooth, respectively. Movement of markers was repeatedly recorded by 6 video cameras from centric occlusion to maximal opening position. Data was analyzed and the movements were constructed in 3D. In order to predict movement (opening, vertical, anterior-posterior) at an incisor-tooth point, multiple regression analysis was performed by making the 27 amounts of 3D movements (X, Y, Z components) of nine facial skin markers as explanatory variables. The difference between the actual value and the predicted value by multiple regressions was calculated, and compared in the 5 ranges of analysis (0-5 mm, 0-10 mm, 0-15 mm, 5 10 mm, 10-15 mm). RESULTS: The movements of mouth opening and vertical direction could be predicted with the accuracy of 0.76 mm (3.7%)and 0.69 mm (4.9%) in the range of 0-15 mm with nine variables, respectively. However, the accuracy of movements in the anterior-posterior direction was 0.40 mm (9.7%) with ten variables. The ratio was larger than these in the movements of mouth opening and vertical direction. CONCLUSIONS: It was suggested that movement of mandibular incisor-tooth point in mouth opening and vertical direction to 15 mm could be predicted from the facial skin markers. PMID- 15818008 TI - [An evaluation of virtual waxing up system with haptic interface]. AB - PURPOSE: The purpose of this experiment was to construct a system of waxing pattern using virtual reality in the odontological field. The success of operation with feedback on the maniphalanx sense of waxing is reported. METHODS: The constructed device was a system based on PHANToM DESKTOP (SensAble Technologies Inc.). This device includes six-degrees-of-freedom input and reproduction of the sense of touch by anti-power feedback. The software which controls this device enables four operations such as "Cutting down", "Piling up", "Melting", and "Finishing" to be applied to the virtual waxing pattern by virtual reality. Finally, the virtual waxing was examined by applying this system to a virtual abutment tooth. RESULTS: With six-degrees-of-freedom input and anti-power feedback, a virtual waxing and the designer's highly developed designs were successfully expressed regardless of the virtual space. CONCLUSIONS: 1. The virtual waxing up could be done by six-degrees-of-freedom input with the stylus in a virtual space. 2. The time taken for all these numerical values of corresponding rate of volume, occlusal, mesio-distal and bucco-lingual views to reach 100+/-5%ranged from 50 to 75 minutes, with an average of 63 minutes. 3. The virtual wax improvement showed the tendency to trace the passage of first arranging the volume, and straightening the outlook on the occlusal view at the end. 4. This system, which provides an interface between man machines, six degrees-of-freedom input and anti-power feedback, has much in common with the existing waxing up and crafting. The demand for special technologies by technical persons and engineers can be kept to a minimum and application of the method can be expanded into education and to objective evaluations. PMID- 15818009 TI - [Verification of precision by 4-dimensional analysis of mandibular movement]. AB - PURPOSE: The 4-dimensional human body model that combines reconstructed 3 dimensional image data (CT and MRI) and exercise data of the human body is widely applied in the medical field. We have applied this technique to assess mandibular movement, particularly in dentistry, and developed a 4-dimensional system of analyzing mandibular movement. The aim of this study was to determine the accuracy of this analysis system. METHODS: A dry skull mandible was fixed at several mandibular positions and the skull-mandible relationship was measured at each position by two procedures: CT scanning was performed and a 3-dimensional image was reconstructed from image data that were regarded as true values; then, optical measurement markers attached to the dental arch were measured 3 dimensionally and calculated as positional measurement values. The measurement values were compared with the corresponding true values to identify measurement errors of the analysis system. RESULTS: The root-mean-square (RMS) errors of the 4-dimensional analysis system of mandibular movement were, on average, 0.41 mm and 0.43 degrees. CONCLUSIONS: These findings suggest that our 4-dimensional analysis system has sufficient measurement accuracy for mandibular movements. PMID- 15818010 TI - [An analysis of angles related to gingival cone and marginal gingiva in natural dentition]. AB - PURPOSE: There are no reports on numerical investigation of suitable shapes of restoration. This study examines the relationship between gingival cone and marginal gingiva in natural dentition, and discusses the standard angles for giving a suitable shape at the gingival cone of restoration. METHODS: Twenty dental casts of healthy subjects with normal dentition were examined. The angles between gingival cone and marginal gingiva (CG angle), and the angles between gingival cone and the long axis of the tooth crown (CA angle) were measured. RESULTS: The mean CG angles of each site were measured between 96.7 degrees (palatal surface of the upper first molar) and 123.9 degrees (labial surface of the lower central incisor). Within the same kind of teeth, lower significant differences were observed among upper and lower, or buccal and lingual surfaces. The CG angles of anterior teeth were 9.5 to 19.2 degrees larger than those of posterior teeth. The CA angles of anterior teeth were negative, and the apex of gingival cone of anterior teeth was directed toward the incisal edge. Conversely, the CA angles of posterior teeth were positive, and the apex was directed toward the root apex. Significant negative correlation was found between CA angle and CG angle (p<0.001). CONCLUSIONS: These results suggested that the relationship between gingival cone and marginal gingiva is fairly constant throughout their angles, and that the cervical shape could affect the shape of marginal gingiva. PMID- 15818011 TI - [The effect of woven glass fiber reinforced composites on the transverse strength of three denture base resins]. AB - PURPOSE: This study evaluated the effect of woven glass fiber reinforced composite (FRC) on transverse strength of denture base materials. METHODS: Three denture base resins (heat-activated resin, autopolymerizing resin and microwave activated resin) and FRC (Vectris Frame, Ivoclar-Vivadent) were selected for the study. The FRC was light-cured and sandwiched in the middle of denture base resin and then the specimen was polymerized according to manufacturers' instructions in a gypsum mold with a cavity of 65 mm x 10 mm x 2.5 mm. Each specimen was immersed in 37 degrees C distilled water for 50 hours. The control group was without FRC. Ten specimens were fabricated for each group. The transverse strength test was performed on a universal testing machine at a crosshead speed of 5 mm/min. RESULTS: The transverse strengths of all denture base resins reinforced with FRC were significantly higher than that of denture base resin bulk. CONCLUSIONS: Within the limitation of the current study, the FRC has an effect upon reinforcement of denture base resin. PMID- 15818012 TI - [Manufacture of a titanium full crown by CAD/CAM--tolerance of the cutting tool]. AB - PURPOSE: We investigated the durability of the cutting tool(bar)that is used when creating a dental crown by using the CAD/CAM system of a computer-assisted NC machine tool. METHODS: We used the chamfer type as the shape of the dental cervical margin of the support tooth model. We also used a mold model whose shaft surfaces on both sides had an 8-degree taper. We used the CAD/CAM system of DENTAL Cadim (Advance Inc.). First, we manufactured a nonocclusion dental crown of the resin-pattern on top of the support dental model, and measured its 3 dimensional shape. Upon measuring the 3-dimensional shape, we created a pure titanium block by using cutting bars continuously for rough processing and for finishing processing separately. Upon completion of the pure-titanium block, we measured the time required for each processing and measured the surface roughness after each processing. RESULTS: The surface roughness Ra of the rough processing became significantly worse after the fifth processing (p<0.01). Rmax became significantly worse after the sixth processing(p<0.05)and after the seventh processing (p<0.01). The surface roughness of both Ra and Rmax of the finishing processing became significantly worse after the fifth processing (p<0.05) and after the seventh processing (p<0.01). CONCLUSIONS: From the viewpoint of quality of the crown's finished surface and compatibility fitness, the number of cuttings, the number of rough processing and the number of finishing processing using the same cutting tool for making a full crown of a molar tooth should be limited to four times. PMID- 15818013 TI - [Influence of cavity lining on marginal leakage beneath cast restoration]. AB - PURPOSE: This study investigated the effect of a cavity lining on marginal leakage of a cast restoration. METHODS: MOD cavities were prepared on human extracted upper third molars. Additional hemispherical cavities (1.6 mm diameter), simulating deep caries, were prepared on axial walls, and they were restored with lining materials including a glass-ionomer, a resin-modified glass ionomer, and a low viscosity composite for a lined group. Metal inlays were cast with a 12% Au-Ag-Pd alloy and luted with four luting agents including a zinc phosphate(HZ), a conventional glass ionomer (FI), a resin-modified glass ionomer (VL), and an adhesive resin (SB). The specimens were stored in 37 degrees C distilled water for 24 hours and subjected to 5,000 thermo-cycles, and then, dye penetration was evaluated after 24 hr immersion in 2% erythrosine solution. Metal disks were luted to dentin disks, which have the same-sized spherical cavities in the central area, with the same luting agents. Shear bond strengths were evaluated for both lined and non-lined specimens after the same storing and thermo-cycle processes. RESULTS: The inlay specimens luted with SB exhibited no dye penetration, while the specimens luted with HZ presented dye penetration reaching to the pulp chamber. The specimens luted with FI and VL showed moderate dye penetration and the values of dye penetration for the cavity-lined group were significantly less compared to those for the lined group (p<0.05). The shear bond strength of the specimens luted with SB and VL was significantly affected by the lining material. The shear bond strength of the specimens luted with HZ or FI was 0 MPa. CONCLUSIONS: The marginal leakage of a cast restoration is affected by the cavity lining but can be improved by using adhesive resin. PMID- 15818014 TI - [The application of metal framework obturator to a patient with maxillary defect]. AB - PATIENT: A 55-year-old patient with left maxillary defect was referred to the clinic in 1987, and an obturator prosthesis was made, but frequent denture base fracture occurred. In 1998, she came to the clinic complaining of unfitted obturator. A metal framework obturator prosthesis was constructed for her maxillofacial rehabilitation. Since delivery of the prosthesis, normal masticatory and speech function and appearance were successfully achieved. DISCUSSION: Oral functions were improved by the obturator prosthesis, but the remaining part of the obturator was broken and caries of the remaining teeth were observed after 3 years. It was considered that frequent recalls were necessary to keep the remaining teeth and obturator prosthesis in good conditions. CONCLUSION: This article describes the prosthetic rehabilitation of left maxillary defect. The metal framework obturator improved function and esthetics. PMID- 15818015 TI - [A clinical case report of complete dentures using resilient denture liners]. AB - PATIENTS: A man (68 years total edentulous) visited us because of masticatory soreness around the mandible. He also suffered from frequent fracture of the dentures. As the poor fitting and severe absorption of the mandible area was considered to be the main cause of the trouble, soft denture lining materials were used. After trying different types of soft denture liners over the new acrylic base dentures, the dentures were finally relined with fluoroelastomer. DISCUSSION: The condition was good and the patient was very satisfied with the lined dentures. The soft liner contributed to the improvement of the QOL of the edentulous patient. CONCLUSION: The use of soft liner was effective for this difficult edentulous case. This case concerns the guideline for the relining of soft liners. PMID- 15818016 TI - [A case report of treatment by complete dentures for a patient with reciprocal clicking]. AB - PATIENT: This case report describes a treatment method for a 68-year-old female with reciprocal clicking on her left temporomandibular joint. Wearing complete dentures for a long time and her unilateral habitual mastication caused this condition. As a countermeasure, an interocclusal record was taken at a position without clicking and a new complete denture set was fabricated. DISCUSSION: In such a case, we usually expect to adopt a new jaw position by adjusting the old dentures. If the degree of disc displacement or transformation of the disc is severe, mandibular repositioning may be difficult. Furthermore, many aged patients have limitated ability to adapt to a new mandibular position. In this case, the degree of disc displacement was slight, so the patient could accept a new position. CONCLUSION: After wearing the new dentures, her clicking disappeared and MRI showed reduction of the disc. She was able to accept the new position. PMID- 15818017 TI - [A case of occlusal reconstruction for a patient with severe chronic periodontitis]. AB - PATIENT: 65 years old, female. Chief complaint:Esthetic problem of upper right incisor because of labial displacement. After initial periodontal therapy, a hawley bite plane was inserted to reposition the mandible and retract the anterior teeth. The anterior teeth were stabilized with pin ledge materials. DISCUSSION: For a patient with severe chronic periodontitis, it is difficult to decide whether to replace or retain the remaining teeth. The hawley bite plane is effective in these cases in order to control occlusal trauma, to reposition the mandible, and to reduce teeth movements. CONCLUSION: It is critical to evaluate the prognosis of each tooth through initial periodontal diagnostics in cases of severe chronic periodontitis. A treatment plan can then be created which will address all known etiologic factors. PMID- 15818018 TI - Malnutrition in the developing world: the lack of food scenario. PMID- 15818019 TI - Are older people starving to death in a world of plenty? PMID- 15818020 TI - Physiological vs. pathological changes of nutritional status over life time. PMID- 15818021 TI - Home enteral nutrition. Demographics and utilization in the United States. PMID- 15818022 TI - Home enteral nutrition. Epidemiology and legislation in Europe. PMID- 15818023 TI - When does malnutrition become a risk? PMID- 15818024 TI - What are the goals of nutritional support? The example of home enteral nutrition. PMID- 15818025 TI - Oral protein and energy supplementation in older people: a systematic review of randomized trials. PMID- 15818026 TI - Efficacy of enteral and parenteral nutrition in cancer patients. PMID- 15818027 TI - Ethics and economics in nutritional support. PMID- 15818028 TI - Pathophysiology of weight loss in older persons. PMID- 15818029 TI - Interaction between nutrition, intestinal flora and the gastrointestinal immune system. PMID- 15818030 TI - Psychoimmunology of nutrition. PMID- 15818031 TI - How can we impact the immune system with pre- and probiotics? PMID- 15818032 TI - How can we modulate cytokine production and action? PMID- 15818033 TI - How can we improve functional outcomes? PMID- 15818034 TI - Withdrawal of warfarin prior to a surgical procedure: time to follow the guidelines? AB - BACKGROUND AND OBJECTIVE: Patients with cardiogenic sources of embolism may be at increased risk of cerebral infarction when anticoagulation therapy is suspended for surgical procedures. The purpose of this study was to determine frequency of cardioembolic cerebral infarction during periprocedural warfarin withdrawal. METHODS: Retrospective analysis of prospective cerebral infarction registry data from two tertiary medical centers. RESULTS: Over a 12-month period, 14 cases of cardioembolic cerebral infarction occurring during the period of warfarin withdrawal for a medical procedure were observed, accounting for 7.1% of the 197 cardioembolic cerebral infarctions encountered. Across all patients, cerebral infarctions developed an average of 5.4 days after the last dose of warfarin (range 3-8). Among the 14 patients (8 males and 6 females) with warfarin cessation-related infarcts, age ranged from 54 to 91 years. Each had been on chronic anticoagulation with warfarin for more than 1 year. Retrospective analysis suggested that all these cerebral infarctions had been potentially preventable. In each case, either the planned procedure did not require discontinuation of warfarin or, when withdrawal was required, no bridging, parenteral anticoagulation was provided to lessen the risk during the warfarin free period. CONCLUSION: Patients at high risk of cardioembolic cerebral infarction may benefit from more intensive management strategies to reduce cerebral infarction risk during periprocedural periods. PMID- 15818035 TI - Hemoglobin-based oxygen carrier distribution inside vascular wall and arterial pressure evolution: is there a relationship? AB - The hemoglobin-based oxygen carriers (HBOC), like dextran-benzene tetracarboxylate-hemoglobin (Dex-BTC-Hb), which are present at high concentrations in plasma disturb arterial pressure and induce hypertension. To study if the increase of mean arterial pressure (MAP) is due to the presence of cell-free hemoglobin (Hb) inside abdominal aortic wall, we followed on a model of 50% isovolemic exchange transfusion (IET) in anesthetized guinea pigs, the kinetic of Dex-BTC-Hb distribution inside abdominal aortic wall and we investigated the relationship between arterial pressure modifications and modified Hb distribution. The administration of Dex-BTC-Hb induced instantaneously an increase of MAP that reached its maximum (53% of hypertension from baseline) at 17 min after the end of the IET and was maintained maximally up to 30 min. A significantly decrease of MAP (45% of hypertension from baseline) was observed after 60 min and the baseline level was recovered at 180 min. The investigation of tissue at 17 min by confocal microscopy showed the presence of free Hb in or upon endothelial cells (EC) in intima and in vasa vasorum. At 180 min, the free Hb was found in or upon EC and inside all abdominal aortic wall meanwhile MAP recovered its basal value. These results suggest for the first time that Hb in intima seems to induce the hypertension observed upon IET but can not sustain it even if Hb stayed present in intima and in abdominal aortic wall. PMID- 15818036 TI - Improved ventricular function during inhalation of PGI(2) aerosol partly relies on enhanced myocardial contractility. AB - Inhaled prostacyclin (PGI(2)) aerosol induces selective pulmonary vasodilation. Further, it improves right ventricular (RV) function, which may largely rely on pulmonary vasodilation, but also on enhanced myocardial contractility. We investigated the effects of the inhaled PGI(2) analogs epoprostenol (EPO) and iloprost (ILO) on RV function and myocardial contractility in 9 anesthetized pigs receiving aerosolized EPO (25 and 50 ng.kg(-1).min(-1)) and, consecutively, ILO (60 ng.kg(-1).min(-1)) for 20 min each. We measured pulmonary artery pressure (PAP), RV ejection fraction (RVEF) and RV end-diastolic-volume (RV-EDV), and left ventricular end-systolic pressure-volume-relation (end-systolic elastance, E(es)). EPO and ILO reduced PAP, increased RVEF and reduced RVEDV. E(es) was enhanced during all doses tested, which reached statistical significance during EPO(25 ng) and ILO, but not during EPO(50 ng). PGI(2) aerosol enhances myocardial contractility in healthy pigs, contributing to improve RV function. PMID- 15818037 TI - Thoracoscopic decortication: first-line therapy for pediatric empyema. AB - Video-assisted thoracoscopic surgery (VATS) has become a popular and well-refined procedure. It has several advantages over open thoracotomy in terms of invasiveness and cosmetics. The aim of this study is to consider if VATS can serve as primary management for pediatric empyema. Between October 2000 and December 2002, 13 children with empyema receiving VATS were retrospectively reviewed. Of them, 5 had tube thoracostomy before VATS intervention (group T), and 8 had VATS as the initial treatment (group V). Their mean (+/- SD) age of groups V and T were 5.75 +/- 4.43 and 4.0 +/- 1.58 years, respectively. The children of group V had a shorter length of hospital stay (group V 10.7 +/- 3.54 days; group T 28.2 +/- 8.32 days), a shorter period of chest tube drainage (group V 5 +/- 1.87 days; group T 25 +/- 11.08 days), less transfusion (group V 0.4 +/- 0.17 units; group T 0.9 +/- 0.27 units) and less radiation exposure (group V 10.3 +/- 3.49; group T 23.4 +/- 11.64). No mortality was noted in the entire series. We conclude that primary VATS is a safe, effective and definitive method which can serve as first-line therapy for children with empyema. PMID- 15818038 TI - Serum procalcitonin levels are elevated in esophageal cancer patients with postoperative infectious complications. AB - BACKGROUND: The normal systemic inflammatory response to surgical stimuli often makes early diagnosis of postoperative infections difficult. PURPOSE: We investigated whether serum procalcitonin (PCT) levels may be a useful marker of bacterial infections in patients after invasive surgery. SUBJECTS AND METHODS: The subjects were 40 patients who had undergone radical surgery for esophageal carcinoma by a right thoracoabdominal approach. Nine patients were diagnosed to have a postoperative infection during the first 7 days after surgery. Changes in serum PCT levels were compared between the group diagnosed to have postoperative infection (infection group) and the group without infection (noninfection group). RESULTS: The postoperative serum PCT levels were significantly higher in the infection group than in the noninfection group (ANOVA: p < 0.01). Serum PCT peaked on postoperative day (POD) 5 in the infection group (8.7 +/- 8.2 ng/ml, mean +/- SD) and on POD 1 in the noninfection group (0.5 +/- 0.5 ng/ml). No significant differences were found between the two groups in leukocyte count, serum CRP or cytokine levels. The receiver operating characteristics (ROC) curve was constructed for infection identification. The area under the ROC curve for peak postoperative PCT was 0.968, and at a cutoff value of 2.0 ng/ml, the sensitivity was 89% and the specificity was 93%. CONCLUSIONS: Serum PCT levels may be useful for the early diagnosis of postoperative infectious complications. PMID- 15818039 TI - Clinicopathological study of cholelithiasis following gastric cancer surgery. AB - The purpose of this study was to compare liver function tests preoperatively and postoperatively in 2 cohorts of patients, those that developed gallstones after gastrectomy for cancer and those that did not develop gallstones. The cohorts were taken from 698 consecutive patients who underwent curative gastrectomy for gastric cancer between April 1980 and March 1995. In comparison with 698 patients, the gallstone group was characterized by the findings that the incidence of cholelithiasis was significantly higher in totally-gastrectomized patients and patients with upper stomach cancer. Comparison of the perioperative status revealed a significantly higher rate of complications in the gallstone patients. Analysis of the changes in liver function showed significantly higher values of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and alkaline phosphatase at time points of 1 and 3 months after surgery in the gallstone patients. The results suggest that the postoperative development of liver dysfunction and complications is associated with the formation of gallstones. PMID- 15818040 TI - Preclinical application of radioimmunoguided surgery using anti-carcinoembryonic antigen biparatopic antibody in the colon cancer. AB - Radioimmunoguided surgery (RIGS) has been known as a sophisticated tool to detect micrometastasis intraoperatively. A preclinical model of RIGS was designed to test the possible clinical applicability of the biparatopic antibody in detecting colorectal cancer. The biparatopic antibody was constructed using two anti carcinoembryonic antigen (CEA)-specific antibodies, T84.66 and PR1A3, reacting against two different epitopes. (125)I-labeled biparatopic antibody was introduced via the principal colonic arteries at the end of operation in 10 operable patients with colon cancer. After 24 h, the radioactivities of the tumors and lymph nodes were counted using the gamma-detecting probe. The radioactivity count was performed ex vivo. The accurate detection in the primary tumors and metastatic lymph nodes were 100 and 88.7% respectively. False-positive detections occurred in 24 of 256 lymph nodes (9.4%), whereas false-negative detections occurred in 5 of them (2%). The most frequent cause of false-positive detection was dissociated radionuclides trapped in the lymphatic tissues. False negative detections occurred mainly from weak targeting by radiolabeled antibody, probably due to weak expression of tumor CEA. Conclusively, as most detection errors appear to be reduced within 3 days in vivo, the biparatopic antibody can efficiently be applied to the clinical RIGS, thereby facilitating accurate detection and removal of occult cancer foci in colorectal cancer. PMID- 15818041 TI - Influence of gadolinium-induced kupffer cell blockade on portal venous tolerance in rat skin allograft transplantation. AB - BACKGROUND: Intraportal injection of donor antigens delays rejection of allografts (portal venous tolerance). The study aimed to investigate the possible influence of prior gadolinium chloride (Gd)-induced Kupffer cell blockade on tolerance to non-vascularized skin allografts induced by means of donor-specific intraportal blood transfusion. MATERIALS AND METHODS: Wistar rats (n = 10) were used as donors and Sprague-Dawley rats (n = 70) as recipients of a non vascularized skin graft. Recipients were divided into groups according to the manipulations prior to transplantation, as follows: (1) no manipulation; (2) donor-specific intrajugular blood transfusion; (3) donor-specific intraportal blood transfusion; (4) Gd administration and donor-specific intrajugular blood transfusion; (5) Gd administration and donor-specific intraportal blood transfusion; (6) Gd administration, and (7) intraportal saline infusion. In a first set of experiments, these manipulations were performed once. In a second set of experiments, the same manipulations were performed twice. Skin allograft was performed 7 days after the last manipulation in all groups. RESULTS: Group 3 showed the highest skin graft survival, particularly after repeated blood transfusion. Graft survival in this group was significantly higher than in any other group. Conversely, group 5 showed the lowest graft survival, particularly after repeated blood transfusion. Graft survival in this group was significantly lower than that of groups 1, 2, 3 and 7. CONCLUSIONS: In this model of skin allograft transplantation, Gd administration abrogates and can even reverse the tolerogenic effect of repeated donor-specific intraportal blood transfusion. PMID- 15818042 TI - Evaluation of sympathetic blockade after intrathecal and epidural lidocaine in rats by laser Doppler perfusion imaging. AB - The widespread use of neuraxial anaesthesia increases the need for animal models to evaluate therapeutic prospects, mechanisms and risks of this technique. As a methodological prerequisite, we characterised the sympathetic blockade after different modes of neuraxial anaesthesia with regard to segments supplying the splanchnic region. Under haemodynamic monitoring, lidocaine 2% or saline were infused via intrathecal (10 microl), lumbar epidural (10 and 30 microl) or thoracic epidural (10 and 30 microl) catheters. Segmental spread of neuraxially infused local anaesthetic was assessed using methylene blue. Mean arterial blood pressure decreased more severely after neuraxial lidocaine in thoracic epidural (10 and 30 microl) compared to high-volume (30 microl anaesthesia animals. Determination of the sympathetic blockade by means of laser Doppler perfusion imaging was restricted to the paws due to a higher density of subcutaneous blood vessels as compared to the abdominal wall (mean +/- SD: 3.93 +/- 0.06 vs. 1.35 +/ 0.05/384 mm(2), p < 0.05). Only high-volume (30 microl) lumbar and thoracic epidural anaesthesia (10 and 30 microl) increased skin perfusion in both hind and front paws. This extensive sympathetic blockade was demonstrated to include splanchnic segments using thermography. Segmental spread of methylene blue did not closely correspond to laser Doppler findings and should be interpreted as minimum rather than exact epidural spread of local anaesthetic. PMID- 15818043 TI - Preservation of vein allograft viability during long-term storage. AB - BACKGROUND: The aim of this study was to compare the vein allograft viability following cryopreservation with that remaining after prolonged refrigerated storage. MATERIALS AND METHODS: Great saphenous vein biopsies had been cryopreserved, and the samples were divided into two matched groups and stored in tissue culture medium for 42 days at +4 degrees C, either with or without regular medium replacement. Each vein allograft was biopsied and assayed for viability every third day by the methyltetrazolium reduction assay. Viability indexes of vein allografts harvested from brain-dead multi-organ donors and from cadavers whose warm ischemic periods were maximally 24 h were also compared. RESULTS: Vein allografts stored for 42 days at +4 degrees C showed a similar viability (58.9 +/ 1.2%) to that of cryopreserved veins (59.7 +/- 2.3%). This was true even when cryopreserved and thawed allografts were subjected to 3 days of post-thaw incubation under presumably favorable conditions (58.7 +/- 1.6%). There was no viability index difference between the samples with medium replaced and not replaced and the specimens harvested from two different donor groups. CONCLUSIONS: Long-term storage of vein allografts at +4 degrees C is a valuable option for regular banking practice. Sufficient amounts can be procured from cadavers similar to tissue donors. PMID- 15818044 TI - Effect of a matrix metalloproteinase activity and TNF-alpha converting enzyme inhibitor on intra-abdominal adhesions. AB - BACKGROUND: Formation of intra-abdominal adhesions depends, in part, on the activity of serine proteinases. Matrix metalloproteinases (MMP) are required for epithelialization of skin wounds but their involvement in mesothelialization of peritoneal wounds and in adhesion pathogenesis is not known. Early tumor necrosis factor-alpha (TNF-alpha) levels have been proposed to reflect propensity to adhesion formation. OBJECTIVE: The impact of MMP activity and secreted TNF-alpha on peritoneal adhesion formation and healing was investigated through systemic administration of the synthetic broad-spectrum MMP and TNF-alpha-converting enzyme (TACE) inhibitor GM 6001. METHODS: Female Sprague-Dawley rats of 4-6 weeks of age were injected subcutaneously daily with GM 6001 100 mg/kg (n = 12) or vehicle (n = 10) starting two days before surgery. In each rat, two standardized peritoneal wounds, 20 mm x 5 mm, were made. One peritoneal wound was sutured whereas the contralateral wound healed by secondary intention. Adhesion formation and peritoneal healing, cell proliferation, and hydroxyproline concentrations were evaluated on postoperative day 7. RESULTS: Total serum TNF-alpha levels increased in vehicle-treated rats (p = 0.019) while GM 6001 treatment effectively prevented the rise in the postoperative phase (p < 0.001). No significant differences were observed in the extent of adhesion formation (p = 0.67) between control (65.0%) and GM 6001-treated (61.5%) animals, or peritoneal wound healing or cell proliferation. Hydroxyproline levels increased in the wounds (p = 0.014) but were not different between the two groups (p = 0.14). CONCLUSIONS: Lack of a striking effect of the MMP and TACE antagonist GM 6001 on postoperative adhesions suggests that MMP activity and TNF-alpha might not be major adhesiogenic factors. PMID- 15818045 TI - Thoracic splenosis twenty-nine years after traumatic splenectomy mimicking intrathoracic neoplasm. AB - Thoracic splenosis refers to a condition of ectopic splenic tissue in the thoracic cavity. It is usually a consequence of splenic tissue seeding in the pleural cavity after thoracoabdominal trauma. A rare case of thoracic splenosis, in a 62-year-old man who had had a traumatic splenectomy due to thoracoabdominal trauma 29 years earlier, is reported. The patient, a heavy smoker, was admitted for evaluation of a left-side thoracic lesion discovered on a plain chest film. Bronchoscopy, CT scan and needle biopsy proved inconclusive for the diagnosis. Exploratory thoracotomy was necessary to establish the diagnosis. During the operation, a thoracic splenosis was confirmed. To date, only 28 cases of thoracic splenosis have been reported in the literature. The purpose of this report is to present a new case of splenosis of the thoracic cavity simulating intrathoracic neoplasm. PMID- 15818047 TI - [Obstetric/Prenatal ultrasound: blessing or burden?]. AB - Sonography has led to tremendous advances in prenatal care, but these advances have given rise to expectations that cannot always be fulfilled. In consequence, women who hae had children with malformations not detected antenatally have sought legal recourse. By establishing a system of training, certification and quality control at European level, physicians can actively contribute to the advancement of our field while limiting the potential for unnecessary problems. PMID- 15818048 TI - [Significance and limitations of obstetrical ultrasound]. AB - Obstetrical ultrasound is practised in Germany, Austria and Switzerland as a screening examination for all pregnancies. Although the reasons for its introduction in Germany in 1981 were dating of pregnancy, detection for multiple pregnancies and detection of intrauterine growth retardation, its aims has shifted towards the detection of fetal malformations when the 3-point obstetrical screening was introduced in 1995. To produce a realistic quality for this large number of examiners, a multi-level system was introduced. Today, contents and quality requirements of the 3 screening examinations (at 10/20/30 weeks), the 3 levels of examination and of the ultrasound equipment can be defined. This should help optimize the still insufficient quality of obstetrical ultrasound and reduce unrealistic expectations. PMID- 15818049 TI - [The development of obstetric ultrasound in Switzerland]. AB - Ultrasound has conquered obstetrics during the last 40 years. Today it is an integral part of antenatal care. Its broad use as a screening method has pushed critics who found open doors at health authorities facing short resources. In Switzerland in early 1996, routine ultrasound as a health technology was temporarily excluded from reimbursement by the health insurances. Under the pressure of the public, the health authorities had to reintroduce reimbursement within a few months. However, reimbursement was linked with several conditions: the ultrasound examination has to be performed by physicians with adequate training and experience; routine ultrasound needs a strict informed consent, and its benefit has to be evidenced. This decision has had a positive impact on quality. After 7 years, Switzerland has a good training program; guidelines for prenatal ultrasound already exist in their second edition, and spot checks of performance showed that quality in Switzerland meets international standards. Ultrasound mainly has a positive cost-effectiveness ratio due to the detection of fetal malformations with consecutive termination of pregnancy. Since termination of pregnancy is ethically questionable, the discussion with respect to reimbursement will most probably go on. In this light, a comprehensive informed consent of the pregnant women is essential. PMID- 15818050 TI - [Training, practice and the increasing dominance of legal aspects in ultrasound in obstetrics and gynecology in Austria]. AB - During the past few years, the Austrian Supreme Court has hanged down two landmark rulings concerning the use of ultrasound in obstetrics and gynecology. After a woman had died of ovarian carcinoma, her family sued the doctor for not having diagnosed the carcinoma by transvaginal ultrasound she had performed 8 months before the tumor was diagnosed. The plaintiffs won the case at the supreme court level, this making Austria the first country with implicit court-ordered ovarian cancer screening. A wrongful life case concerning a baby that had been born with bone deformities was also decided in favor of the plaintiffs, declaring ultrasound in pregnancy to be an instrument intended to help women decide whether or not to have an abortion. The impact of these two cases on the teaching and clinical application of ultrasound in obstetrics and gynecology in Austria is discussed. PMID- 15818051 TI - [Screening of chromosome anomalies during the first trimester]. AB - In view of today's knowledge, it is evident that a very efficient screening for chromosome anomalies can be carried out during the first trimester. Prospective studies of a total of 200,868 pregnancies-among them 871 fetuses with trisomy 21 have shown that measuring the nuchal transparency can identify 76.8% of fetuses with trisomy 21, with a false-positive rate of 4.2%. If the measurement of nuchal transparency is combined with that of the maternal serum concentrations of free human beta-choriogonadotropin and pregnancy-associated plasma A, the detection rate is 87.0% with a false-positive rate of 5% (prospective studies of altogether 44,630 pregnancies with 215 fetuses suffering from trisomy 21). At present, further signs of Down syndrome in the first trimester are being investigated, such as the missing fetal nasal bone, the maxilla and the blood flow pattern in the ductus venosus. Well-known signs of trisomy 13 and 18, which are already visible in the first trimester, are megacystis, omphalocele, polydactyly and holoprosencephaly. Most pregnant women prefer being screened during the first instead of the second trimester. Therefore every expectant mother should be offered an appropriate examination during the first trimester. It is essential for the effectiveness of the screening that the examiners be suitably trained and that the results of the ultrasound and laboratory examinations be subjected to a regular external quality control. In Austria, there is a general consent to follow the guidelines of the Fetal Medicine Foundation. PMID- 15818052 TI - [Are the swiss dying out? Demographic facts and perspectives concerning the current situation of child-bearing in Europe]. AB - In the last decades, the birthrates in most European countries have fallen considerably below the target value needed for the maintenance of a constant population. In Switzerland, the total fertility is 1.37 children per woman between 15 and 48 years of age (target 2.1), and in Swiss women along-excluding foreigners-it has reached the historical low of 1.21 children. The birth excess of the native population has been negative since 1998, which means that the number of inhabitants in Switzerland is only kept constant by immigrants. If the low birthrate continues for another 200 years, the native population-without immigration-will melt down to 2% of today's size, in other words: it will become extinct. The reasons for this deplorable evolution and possible countermeasures will briefly be discussed. PMID- 15818053 TI - [Morbidity of dilatation and evacuation in the second trimester: an analysis]. AB - OBJECTIVE: To compare the rates of complications of dilatation and evacuation (D&E) in the second trimester of pregnancy. STUDY DESIGN: Retrospective analysis of early complications after D&E in the second trimester (group A: 1988-1994) in comparison with D&E in the first trimester (group B: 1997) and with the induction of abortion by extra-amniotic application of prostaglandins in the second trimester of pregnancy (literature). RESULTS: The overall complication rate of D&E was statistically significantly higher in the second than in the first trimester (p < 0.0001). The most frequent complication in both groups was a blood loss > 500 ml (p < 0.009). As with blood loss, there was a statistically significant increase in the incidence of postoperative bleeding (p < 0.001) and fever > 38 degrees C (p = 0.042) with rising gestational age, too. We registered similar rates for incomplete abortion, infection, cervical injury, uterine perforation and thrombosis. CONCLUSION: Second-trimester pregnancy termination with D&E is associated with higher morbidity rates than in the first trimester. Overall, the rates of complications of D&E in our study were acceptable and comparable with the results of previous studies. Compared with the induction of abortion by extra-amniotic prostaglandins, D&E shows lower morbidity rates. In particular, the advantages of D&E are in the early second trimester. PMID- 15818054 TI - [Sentinel lymphonodectomy in breast cancer - our own experience. Part 2]. AB - The aim of this investigation was to document the personal learning curve of the sentinel lymphonodectomy in breast surgery and to compare it with the experiences of other authors. RESULTS: Between July 1999 and May 2004, the authors performed 218 sentinel lymphonodectomies, 48 of them during the 24-month evaluation period with consecutive full axillary dissection. The overall detection rate was 212/218 or 97.2%, while during the evaluation period it was 38/48 or 89.6% (failures were procedures No. 6, 7, 22, 24 and 47). Among the first 48 cases, 24 had histologically proven metastasis to one or more of the axillary lymph nodes. In 3 of these cases, we were not able to detect a sentinel lymph node, and 4 times the sentinel lymph node was false-negative after rapid section. The false-negative results during the learning period belonged to cases No. 2, 10, 23 and 29 (4/21 or 19%). From September 2001 until May 2004, the detection rate was over 99% (165/166). DISCUSSION: Our own experience documents the initial difficulties and insecurities with this operative procedure. The common recommendations, i.e. that sentinel lymphonodectomy without axillary clearance should not be offered before having completed one's own learning curve with the aid of experienced surgeons, are to be supported. PMID- 15818055 TI - Neonatal screening for congenital adrenal hyperplasia in North-Eastern Italy: a report three years into the program. AB - AIMS: To evaluate the incidence of congenital adrenal hyperplasia (CAH) in the Northern Italian population and the efficiency of the North-Eastern Italy screening program. To adjust cut-off levels for 17-hydroxyprogesterone (17-OHP) in relation to gestational age and birth weight, comparing the benefits in terms of reduction of recall rates with the two approaches and ultimately choosing the better of the two. SUBJECTS AND METHODS: Since September 2001, blood samples from neonates born in North-Eastern Italy have been screened with a fluoroimmunoassay method for 17-OHP determination (DELFIA). A preliminary cut-off level of > or = 30 nmol/l was set both for term and preterm newborns. The values of 17-OHP were analysed using statistical methods in relation to gestational age and birth weight in order to modify the cut-off on the basis of our data. RESULTS: After 33 months of screening we screened 128,282 newborns and detected 6 affected babies. During the first 8 months of screening among the recalled babies, 89.6 and 78.1% were preterm and low-birth-weight newborns, respectively, with a recall rate of 2.59% for premature neonates and of 4.94% for babies with birth weights < 2,500 g. We chose a new cut-off value of 50 nmol/l for preterm newborns only and, after 4 months, the recall rate was reduced to 0.83% for these infants and to 1.83% for low-birth-weight infants. CONCLUSION: After 33 months of screening for CAH in North-Eastern Italy, we report an incidence of 1:21,380. In 5 out of 6 affected babies, the diagnosis was established only after a positive screening test, which prevented a severe salt-wasting crisis in these babies. The cut-off level related to gestational age led to a significant reduction in the number of false positives among preterm babies.We therefore intend to continue with the screening program for CAH in North-Eastern Italy, keeping a gestational-age-related cut-off in the hope that our data may encourage a national screening program for CAH. PMID- 15818056 TI - Urinary excretion of transferrin and orosomucoid are increased after acute protein loading in healthy subjects. AB - BACKGROUND/AIMS: The aim of this study was to elucidate what kind of plasma proteins would change their urinary excretions when the glomerular filtration rate (GFR) was increased. METHODS: We measured urinary excretions of three plasma proteins with different molecular radii (MR) and isoelectric points (pI): albumin, orosomucoid (OM) and transferrin (Tf), after acute protein loading in healthy subjects. RESULTS: Urinary excretion of OM with more anioic charge and smaller MR than albumin, and Tf with more cationic charge and slightly larger molecular weight than albumin, significantly increased in parallel with increased creatinine clearances after acute protein loading. These renal responses returned to basal levels 9 h after protein ingestion. In contrast, increases in urinary excretion of albumin were not observed. CONCLUSION: Because these findings could not be explained by changes in either size or charge selectivity of shunt pores in the glomerular capillary wall, it is suggested that urinary excretion of albumin may have a special property that distinguishes it from other plasma proteins and may be a less sensitive marker to reflect changes in renal hemodynamics than the other plasma proteins. PMID- 15818057 TI - Influence of intravenous drug abuse on vascular access placement and survival in HIV-seropositive patients. AB - BACKGROUND: The influence of intravenous drug abuse (IVDA) on hemodialysis access placement practices and access survival in HIV-infected patients is unknown. METHODS: We conducted a retrospective study of 60, HIV seropositive, maintenance hemodialysis patients. Type of access and assisted access survival (measured from date of placement) were compared in those with (77%) and without (23%) a history of IVDA. RESULTS: Mean age was 37.8 years, mean baseline serum albumin was 2.9 g/dl and median CD4 count was 222 cells/mm3. Fifteen patients, all IVDA, were dialyzed using only tunneled catheters (median number of catheters per person (range): 2.5 (1-11)). There were longer delays in creation of a permanent access (p = 0.08), but no difference in the type of permanent access placed in IVDA versus the non-IVDA group. Over 1,051 cumulative months of access follow-up, 134 tunneled catheters, 28 grafts and 19 fistulae were placed, with observed failure rates of 1 per 4.7 access-months, 1 per 19.7 access-months, 1 per 38.2 access months, respectively. The adjusted relative hazard of access failure for grafts versus catheters was 0.41 (95% CI: 0.23, 0.72; p = 0.002) and for fistulae versus catheters was 0.21 (95% CI: 0.08, 0.52; p = 0.001). Thirty-two percent of accesses were removed due to infection, an infection removal rate for catheters of 1 per 7.8 access-months and for grafts of 1 per 62.5 access-months; all graft infections occurred in the IVDA group. No fistula was removed due to infection. CONCLUSION: Fistulae are the first line of choice for hemodialysis access in HIV seropositive patients regardless of IVDA history; if not feasible, graft placement in non-IVDA or abstinent IVDA patients is recommended. In those with active IVDA, the optimal method of renal replacement therapy and type of hemodialysis access remain uncertain. PMID- 15818058 TI - Limitations of CA125 as an index of peritoneal mesothelial cell mass. AB - BACKGROUND: CA125 is commonly used as an index of the mesothelial cell mass in patients treated with peritoneal dialysis. However, we have no data that show a direct relationship between the number of mesothelial cells, their functional properties, and the amount of CA125 produced in these cells. METHODS: Experiments were performed on primary in vitro cultures of human peritoneal mesothelial cells obtained from 32 donors of various ages and of both sexes. Spontaneous release of CA125 from the confluent mesothelial cells was measured and correlated with the number of cells in monolayers and with their functional properties. We also studied acute effects of cytokines (IL-1beta, TNF-alpha, and INF-gamma) and the chronic effects of glucose (45 mM) on the CA125 content in mesothelial cells and the release of this antigen from their cytosol. RESULTS: Cells from older donors released more CA125, but we found no correlation between the number of cells and the amount of CA125 released from their cytosol. The synthesis of CA125 in mesothelial cells does not correlate with the amount of monocyte chemoattractant protein 1 or interleukin-6 produced in these cells. Acute exposure to cytokines did not modify CA125 content or its release from mesothelial cells. Chronic exposure of mesothelial cells for 4 weeks to glucose (45 mM) decreased the CA125 content of their cytosol and the release of this antigen into the culture medium. Mannitol, at the same concentration and under the same conditions, did not produce these effects, namely a decrease in the CA125 content in the cytosol or its release into the cultural medium. CONCLUSIONS: The amount of CA125 released from mesothelial cells is not a good index of their number or their functional properties, because the CA125 release depends not only on the number of cells, but also on their properties. Furthermore, the process is affected by the age of the cell donor and environmental factors such as a high glucose content. The results of this study show the limitations of CA125 as an index of the mesothelial cell mass and viability. PMID- 15818059 TI - CA125: holy grail or a poisoned chalice. PMID- 15818060 TI - The synthetic PPARgamma agonist troglitazone inhibits IL-5-induced CD69 upregulation and eosinophil-derived neurotoxin release from eosinophils. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a nuclear receptor that regulates lipid metabolism. Recently, PPARgamma was reported to be a negative regulator in the immune system. Eosinophils also express PPARgamma, however, the role of PPARgamma in eosinophil functions is not well understood. Surface expression of CD69 and eosinophil-derived neurotoxin (EDN) release are well-known activation markers of eosinophils. We investigated the effect of a PPARgamma agonist on human eosinophil functions such as IL-5-induced CD69 surface expression and EDN release. IL-5 significantly induced eosinophil CD69 surface expression analyzed using flow cytometry and EDN release measured by ELISA. IL-5 induced eosinophil CD69 surface expression and EDN release were significantly inhibited by the synthetic PPARgamma agonist troglitazone, and these effects were reversed by a PPARgamma antagonist. The PPARgamma agonist troglitazone has a potent inhibitory effect on activation and degranulation of eosinophils, and it may be a therapeutic modality for the treatment of allergic diseases. PMID- 15818070 TI - Functional capacity testing in patients with chronic pain. PMID- 15818071 TI - The differential influence of pain and fatigue on physical performance and health status in ambulatory patients with human immunodeficiency virus. AB - OBJECTIVES: The purposes of this study were to: 1) characterize physical performance in individuals with human immunodeficiency virus; and 2) examine group differences by pain and fatigue on a multivariate profile of disease, physical, and psychologic symptoms. METHODS: One hundred outpatients, 78 men and 22 women (mean age 40.70 +/- 7.49 years) participated. Patients completed a battery of physical performance tests in which the time taken or the distance reached or walked was measured. Self-report questionnaires included measures of pain (0-10 numerical rating scale), fatigue (Brief Fatigue Inventory), and perceived health status (Medical Outcomes Survey-HIV scale). RESULTS: Physical performance was compromised in a task specific manner. Patients took twice as long as healthy individuals on a belt-tie and 4 times as long on a sit-to-stand task and in 6 minutes walked 75% of the distance covered by healthy individuals. Fifty percent of patients (n = 50) had pain at the time of testing (mean 6.3 +/- 2.4), and 98% had fatigue (mean 5.4 +/- 2.3). Multivariate analysis of variance showed pain had a greater influence on performance than fatigue. Pain, distance walked in 6 minutes, and unloaded forward reach accounted for 26% of the variability in quality of life (r = 0.51, P < or = 0.0001). DISCUSSION: Pain has a substantial impact on physical performance and quality of life among ambulatory human immunodeficiency virus patients. Fatigue also impacts physical performance. Compromised ability to perform certain physical tasks affects quality of life. Further investigation of the roles of these relevant variables should be investigated in path analyses. PMID- 15818072 TI - Functional capacity evaluation as a performance measure: evidence for a new approach for clients with chronic back pain. AB - OBJECTIVES: To report the research and development of a new approach to Functional Capacity Evaluation, the Gibson Approach to Functional Capacity Evaluation (GAPP FCE) for chronic back pain clients. METHODS: Four studies, including pilot and feasibility testing, expert review, and preliminary interrater reliability examination, are described here. Participants included 7 healthy young adults and 19 rehabilitation clients with back pain who underwent assessment using the GAPP FCE. Thirteen therapists were trained in the approach and were silently observed administering the Functional Capacity Evaluations by at least 1 other trained therapists or the first investigator or both. An expert review using 5 expert occupational therapists was also conducted. RESULTS: Study 1, the pilot with healthy individuals, indicated that the GAPP FCE was a feasible approach with good utility. Study 2, a pilot using 2 trained therapists assessing 5 back pain clients, supported the clinical feasibility of the approach. The expert review in Study 3 found support for GAPP FCE. Study 4, a trial of the approach with 14 rehabilitation clients, found support for the interrater reliability of recommendations for return to work based on performance in the GAPP FCE. DISCUSSION: The evidence thus far available supports the GAPP FCE as an approach that provides a sound method for evaluating the performance of the physical demands of work with clients with chronic back pain. The tool has been shown to have good face and content validity, to meet acceptable test standards, and to have reasonable interrater reliability. Further research is occurring to look at a larger interrater reliability study, to further examine content validity, and to examine predictive validity. PMID- 15818073 TI - Working through the pain: a controlled study of the impact of persistent pain on performing a computer task. AB - OBJECTIVES: A large percentage of employees experience persistent pain while at work. This situation can become costly to employers with large amounts of lost production-time, absenteeism, and long-term disability. The link or transition between working through (ignoring) pain and disabling pain is unknown. This paper presents the results of a controlled study examining the impact of persistent pain on performance in a working population. Benefits of early detection are discussed. METHODS: This was a controlled, repeated measures study using 3 types of measures: questionnaires (pain, pain anxiety, daily memory, and attention mistakes); actigraphic monitoring to assure the absence of sleep deprivation; and the Performance Assessment Battery, a computer-based series of tests. Participants were studied during 3 time periods (9:00 AM, 3:00 PM, and 9:00 PM). RESULTS: Forty participants (20 pain, 20 controls) were studied. For all tasks, pain participants were slower than controls with significant findings on 2 tasks and less accurate with significant differences on 1 task. DISCUSSION: Unlike other studies that either induced pain or used persons with complex pain conditions, this study used participants with a low level of pain intensity and had a majority still engaged in full-time employment. Our results found that people with persistent low-level pain demonstrate a reduction in performance compared with controls. Our study revealed that using a sensitive tool to detect minor performance deficits could indicate pain interference. The early detection of pain interference would provide an opportunity for prevention programs to have a pre-emptive effect on work-related musculoskeletal disorders. PMID- 15818074 TI - Self-report and pain behavior among patients with chronic pain. AB - OBJECTIVES: To determine the relationship between pain behaviors and self-report of pain and disability in patients with chronic pain. METHODS: Thirty-nine patients (59% women), aged 19 to 79 years, admitted to a Multidisciplinary Pain Center with chronic noncancer pain, were assessed on the following: 1) pain intensity using a 0 to 10 Numerical Graphic Rating Scale; 2) the Pain Disability Index; 3) the Pain Self-Efficacy Questionnaire; 4) a 10-minute videotaped session involving sitting, standing, walking, and reclining, after which pain behaviors were coded using a standardized observational protocol. RESULTS: Interrelationships among pain intensity, pain disability, self-efficacy, and pain behavior were tested using the Pearson product-moment correlations. Significant relationships were found between pain behavior frequencies and self-reported pain intensity (r = 0.29, P < 0.05), self-reported pain disability (r = 0.54, P < 0.0005), and reported self-efficacy (r = -0.42, P < 0.005). Multiple regression analyses were performed to further investigate these interrelationships. Only the Pain Disability Index score was found to make a significant unique contribution (semipartial correlation of 15%, P < 0.008) to the prediction of total pain behavior score. DISCUSSION: Findings suggest that pain behavior observation is a valid and reliable assessment tool for use with a heterogeneous chronic pain population. Significant associations were found between pain behaviors and self report measures of pain intensity, pain disability, and self-efficacy; pain intensity scores displayed a weak relationship; and pain disability scores the strongest relationship with pain behavior. Viewed with previous research, the results of this study indicate the value of a multimodal, cognitive-behavioral approach to assessing patients with chronic pain. PMID- 15818075 TI - Pain-related factors contributing to muscle inhibition in patients with chronic low back pain: an experimental investigation based on superimposed electrical stimulation. AB - OBJECTIVE: To evaluate muscle strength, as a component of physical deconditioning, and central activation ratio, representing the performance level during testing, in patients with chronic low back pain as compared to healthy controls, and to evaluate the contribution of cognitive-behavioral and pain related factors to the central activation ration of patients with chronic low back pain. METHODS: Twenty-five patients with chronic low back pain and 25 age and gender-matched controls participated. Muscle strength, that is, peak torque of the quadriceps muscle, was measured on a Cybex dynamometer. During peak torque, the quadriceps muscle was percutaneously stimulated using superimposed electrical stimulation, generating an additional twitch torque in case of submaximal performance. The central activation ratio was calculated as peak torque/(peak torque + additional twitch torque). To evaluate cognitive-behavioral and pain-related factors influencing the central activation ratio, measures of fear of injury, pain catastrophizing, psychologic distress, and pain intensity were used. Differences between groups were tested using either T tests or Mann Whitney U tests. Associations were tested by partial correlation coefficients controlling for gender. RESULTS: The male:female ratio was 15:10. Mean age and chronic low back pain duration were 42.7 (+/-9.5) and 9.9 (+/-8.3) years, respectively. Mean muscle torque (per kg lean body mass) in patients (1.95 Nm/kg +/-0.8) was less than in controls (3.16 Nm/kg +/-0.7) (P < 0.01). Median central activation ratio was lower in patients (P < 0.05). Patients experiencing increased psychologic distress and patients with a higher current pain level showed a lower central activation ratio (P < 0.05). CONCLUSIONS: When interpreting decreased muscle strength in terms of physical deconditioning in patients with chronic low back pain, submaximal performance has to be taken into account. The results suggest that patients with chronic low back pain who report increased psychologic distress and a higher level of current pain tend to show increased inhibition of muscle activity, leading to submaximal performance. PMID- 15818076 TI - The time to onset and overall analgesic efficacy of rofecoxib 50 mg: a meta analysis of 13 randomized clinical trials. AB - OBJECTIVE: To determine the time to onset of analgesia of rofecoxib based on a patient-level meta-analysis of randomized, placebo-controlled, postoperative oral surgery pain studies. METHODS: A search on MEDLINE and of Merck data on file was conducted to identify studies that met the inclusion criteria. Meta-analysis inclusion criteria required that patients were treated with a single oral dose of rofecoxib 50 mg when they experienced moderate or severe pain after surgical extraction of > or = 2 third molars; study design involved patient randomization, double-blinding, and matching placebo, and onset data from individual patients were available. The meta-analysis of time to onset also required that studies used the two-stopwatch method. Eleven studies fulfilled the onset criteria and included patients who received a single dose of rofecoxib 50 mg (N = 1220) or placebo (N = 483). These studies were analyzed to determine time to onset of analgesia, time to perceptible pain relief, percentage of patients achieving onset of analgesia, and duration of analgesia. Six of the 11 studies included a nonselective nonsteroidal anti-inflammatory drug (N = 303) and were included in the onset meta-analysis for comparison. The meta-analysis of overall efficacy also required that data on total pain relief scores over 8 hours were available. Over-all effectiveness of analgesia was based on analysis of 13 studies involving 1330 rofecoxib patients and 570 placebo patients on the endpoints of total pain relief scores over 8 hours and patient global assessment of response to therapy at 24 hours. Eight of the 13 studies with a nonselective nonsteroidal anti inflammatory drug comparator (N = 391) were included for the efficacy meta analysis. RESULTS: Patient demographics and baseline characteristics were similar across treatment groups in each study. Median time to onset of analgesia for rofecoxib was 34 minutes (95% CI, 31-38 minutes), significantly faster than placebo, which did not achieve onset within the 4 hours the assessment was conducted (P < 0.001). Duration of analgesia for rofecoxib 50 mg was > 24 hours. Rofecoxib achieved a greater mean total pain relief score over 8 hours than placebo (17.4 versus 4.4; P < 0.001) and a greater patient response rate on patient global assessment of response to therapy at 24 hours than placebo (73% versus 16%; P < 0.001). Outcomes were similar between the rofecoxib group and the nonselective nonsteroidal anti-inflammatory drug group. CONCLUSION: In this meta analysis of over 1200 rofecoxib-treated patients, a single dose of rofecoxib 50 mg demonstrated a rapid onset of analgesia in approximately half an hour combined with sustained effectiveness, supporting its use as a treatment of acute pain. PMID- 15818077 TI - Chronic pain and posttraumatic stress symptoms in litigating motor vehicle accident victims. AB - OBJECTIVE: There has been little research examining chronic pain and posttraumatic stress symptoms in persons injured in motor vehicle accidents. The purpose of this study was to evaluate differences in physical injury and impairment, psychological distress, and pain coping strategies in litigating chronic pain patients low and high in motor vehicle accident-related posttraumatic stress symptoms. DESIGN: A total of 160 consecutive chronic pain patients referred for psychological-legal assessment underwent semistructured interview and testing. The testing battery included the Minnesota Multiphasic Personality Inventory-2, the Multidimensional Pain Inventory, the Sickness Impact Profile, and the Coping Strategies Questionnaire. Using the sample-specific median split of 18 posttraumatic stress symptoms on the Minnesota Multiphasic Personality Inventory-2 Posttraumatic Stress Disorder scale, chronic pain patients were categorized as evidencing low or high levels of posttraumatic stress symptoms. RESULTS: The findings indicate that participants evidencing high posttraumatic stress symptoms had more physical impairment, psychological distress, and maladaptive pain coping strategies and were more likely to be treated with antidepressants, other medications, and psychological management than participants evidencing low posttraumatic stress symptoms. A discriminant function analysis was performed using the full combination of physical injury and impairment, psychological distress, and pain coping variables in the prediction of posttraumatic stress symptom-defined group membership. The resulting discriminant function accounted for 61% of the between-group variance and correctly classified 92% of participants who were low in posttraumatic stress symptoms and 88% of participants who were high in posttraumatic stress symptoms. CONCLUSIONS: Chronic pain and posttraumatic stress symptoms in litigating motor vehicle accident victims are associated with increased physical and psychological morbidity. PMID- 15818078 TI - Pain report and pain-related evoked potentials operantly conditioned. AB - OBJECTIVE: The purpose of the present study was to answer the question whether pain report can be increased and decreased by operant conditioning. We predicted that the conditioned pain effects would remain significant after correction for social desirability and fantasy proneness. Furthermore, we tried to show that the neurophysiologic basis of verbal pain report, defined by pain (event)-related potentials, was affected by the conditioning procedure. Specifically, it was expected that the central recording site N150-P260 pain (event)-related potentials peak-to-peak amplitude would show the largest effect. METHODS: There were 4 groups: an up-conditioning group, a down-conditioning group, an at-random conditioning, and no-feedback control group. Healthy patients received 45 calibrated pain stimuli of equal physical intensity and were asked to rate the pain intensity they experienced. Up-conditioning was established by rewarding the subject if pain report increased compared with the previous trial. Down conditioning of pain report was achieved by rewarding a decrease in the pain score. RESULTS: Results of the subjective pain reports clearly indicated that both forms of conditioning succeeded. Up-conditioning resulted in the highest pain scores and down-conditioning in the lowest scores with the two control groups in between them. Controlling for level of social desirability and fantasy proneness did not negatively influence these results. The N150-P260 pain (event) related potentials peak-to-peak central recording site component also showed the predicted effect and reached statistical significance. DISCUSSION: We concluded that the subjective report of pain as well as a specific pain-related potentials component can be operantly conditioned. PMID- 15818079 TI - Physicians' assessments versus measured symptoms of complex regional pain syndrome type 1: presence and severity. AB - OBJECTIVE: To assess the validity of physician's judgements of symptoms associated with Complex Regional Pain Syndrome Type 1. METHODS: The validity of physicians' judgments was assessed using measurements with regard to presence and severity of pain, temperature and volume asymmetry, and reduction in active range of motion in 66 Complex Regional Pain Syndrome Type 1 outpatients. Measurements were performed using Visual Analog Scales and McGill (number of words chosen total) for pain, infrared thermography for temperature differences, water displacement volumeters for volume differences, and hand-held goniometers for active range of motion. Physicians were blind to the outcomes of the measurements. RESULTS: In general, physicians were capable of determining presence or absence of measured symptoms and indicate the direction of the symptom asymmetry. Establishing presence of temperature and volume asymmetries was, however, inadequate. Poor to moderate correspondence was found for the severity of individual symptoms between physicians' judgments and measurements. For the total number of assessments, correlation coefficients ranged from 0.39 for Volume to 0.68 for Pain. In general, lower correlations and percentages of association for Volume and Temperature were found. Monitoring changes between consecutive patient assessments showed poor correspondence between both assessment methods, with correlation coefficients ranging from 0.25 for Volume to 0.37 for Pain. CONCLUSIONS: We conclude that establishing the presence of Complex Regional Pain Syndrome Type 1 symptoms, except for temperature and volume asymmetries, and monitoring of disease progression based on these symptoms can be performed by clinical judgment. The severity of the individual symptoms evaluated in this study should be measured with reliable and valid measurement instruments. PMID- 15818080 TI - Faces scales for the measurement of postoperative pain intensity in children following minor surgery. AB - OBJECTIVES: Faces scales are commonly used to obtain self-reports of pain intensity from children. Previous research using hypothetical vignettes and pain following venepuncture has found differences in children's pain ratings as a function of the type of faces scale used. The purpose of the present study was to determine whether scales beginning with a smiling rather than neutral "no pain" face would produce higher ratings in the assessment of postoperative pain intensity in children and to compare ratings using different faces scales to those reported with an additional independent measure of pain intensity. METHODS: Participants were 78 children between the ages of 5 and 13 years undergoing surgery, one of their parents, and their postoperative care nurse. Following surgery, children were asked to provide a rating of their current pain intensity using a set of 5 successively administered faces scales and the Colored Analog Scale (CAS). Parents and nurses provided independent ratings using the same measures. RESULTS: Results showed that parents and nurses rated significantly more pain when using scales with a smiling rather than a neutral "no pain" face. This pattern was not as clear for the children's ratings, although their highest ratings were provided when using a smiling "no pain" faces scale. Children's and nurses' ratings on the CAS were generally more similar to their ratings using scales with neutral "no pain" faces, whereas parents' CAS ratings tended to fall in between ratings provided on the smiling and neutral "no pain" faces scales. Scale preference, age and sex differences in pain ratings, and child-parent-nurse agreement in pain ratings are also examined. DISCUSSION: Children's and parents' ratings of postoperative pain intensity are influenced by the presence of a smiling "no pain" face at the beginning of faces scales, with such scales producing significantly higher ratings than scales with neutral "no pain" faces. Ratings on the independent CAS measure were more comparable to those provided on faces scales with neutral "no pain" faces. Nurses are also susceptible to the influencing effect of a smiling face at the beginning of a faces scale. PMID- 15818081 TI - Severe coronary artery spasm can be associated with hyperthyroidism. AB - BACKGROUND: Coronary artery spasm is not infrequently seen in Korea. Most of the patients with coronary spasm show a focal spasm in coronary angiography. However, the cause of the disease is not well known. There have been a few anecdotal case reports of coronary artery spasm associated with hyperthyroidism, but there has not been a report concerning a large series of such patients. Over a period of 5 years and 8 months, we experienced eight patients having the diffuse or severe type of coronary artery spasm in association with hyperthyroidism. METHODS: We investigated the characteristics of the patients with coronary artery spasm, which was diagnosed by coronary angiography or by provocation with an intracoronary injection of acetylcholine or ergonovine. The demographic data, coronary angiographic findings, thyroid function test results, and the follow-up clinical data of the eight patients having coronary artery spasm associated with hyperthyroidism were analyzed. RESULTS: All eight patients had Graves' disease. In six patients, the coronary arterial vasoconstriction developed during the coronary angiography without an injection of ergonovine. In three patients, the left main stem coronary artery was involved in the spasm. Among these eight patients, five were female, and all of these female patients were < or = 51 years old. All of the patients were treated with anti-thyroid medications, calcium channel blockers, and long-acting nitroglycerines; they all remained free of chest pain during the median follow-up period of 5 years. CONCLUSIONS: A severe form of coronary artery spasm could be associated with hyperthyroidism. A high level of suspicion and the thyroid function study should be mandatory for patients with coronary artery spasm, especially for the young female patients. PMID- 15818082 TI - Post-systolic shortening during dobutamine stress echocardiography predicts cardiac survival in patients with severe left ventricular dysfunction. AB - BACKGROUND: Patients with severe left ventricular dysfunction and myocardial viability by dobutamine stress echocardiography (DSE) or F18-fluorodeoxyglucose single-photon emission computed tomography (FDG-SPECT), experience improved survival after coronary revascularization. Pulsed wave-tissue Doppler imaging (PW TDI)-derived ejection phase shortening (EPS) and post-systolic shortening (PSS) velocities may help to quantify DSE. We assessed these variables in a prospective long-term follow-up. METHODS: Eighty patients (58 men, mean age 63+/-9 years) with left ventricular dysfunction (radionuclide ventriculography mean ejection fraction, 34+/-11%) underwent both DSE and FDG-SPECT for myocardial viability. Viable myocardium was improvement from rest to low dose or worsening of wall motion at peak DSE and normal perfusion, mildly reduced perfusion with FDG uptake or severely reduced or absent perfusion with increased FDG uptake (mismatch) at FDG-SPECT. EPS, PSS velocities and EPS/PSS ratio during DSE were analysed using a six-segment model. Coronary revascularization bypass grafting was performed in 62 patients. All patients completed a long-term (9-year) follow-up for cardiac death. RESULTS: The segmental prevalence of severe dyssynergy was 77%. On a patient basis myocardial viability was detected by EPS/PSS ratio (31%), FDG-SPECT (34%) and DSE (26%). A significant improvement of Kaplan-Meier survival was predicted in viable compared with nonviable revascularized patients (P < 0.01). Both EPS/PSS ratio and FDG-SPECT, compared to DSE alone, tended to allocate more accurately univariate prediction of death-free outcome (odds ratio, 2.5 and 2.7 compared with 2.1). CONCLUSIONS: TDI adds objective variables to DSE, helping to recognize viable myocardium and optimize prediction of death-free outcome in long term follow-up, with favorable comparison with nuclear techniques. PMID- 15818083 TI - Tumour necrosis factor-alpha and interleukin-6 release during primary percutaneous coronary intervention for acute myocardial infarction is related to coronary collateral flow. AB - OBJECTIVES: We tested the hypothesis that there was an association between tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) release and measured coronary collateral flow in patients undergoing primary percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI). BACKGROUND: Tumour necrosis factor-alpha and IL-6 increase during acute myocardial infarction (AMI). However, their relation to coronary collateral flow is unknown. METHODS: Twelve patients with AMI due to complete thrombotic coronary occlusion underwent primary PCI within 12 h of symptom onset. Doppler-derived collateral flow index (CFI) was measured during first balloon inflation. TNF-alpha, IL-6, creatine kinase (CK), CK-MB fraction were measured from venous plasma samples serially for 24 h. Area at risk was determined off-line by coronary arteriography. Ejection fraction (EF) was measured using biplane left ventricular angiography. RESULTS: Maximal CK release varied between 569 and 6276 U/l and area at risk varied between 7 and 47% of myocardium. Tumour necrosis factor-alpha (peak 4.4+/-0.5 pg/ml) and IL-6 (peak 35.5+/-3.0 pg/ml) increased in all patients. Peak TNF-alpha and IL-6 release was independent of CK, CKMB. No minimal threshold of myocardial necrosis for cytokine expression could be detected. Similarly, TNF-alpha and IL-6 release was also independent of time to reperfusion, area at risk or EF. Using univariate regression analysis, peak TNF-alpha inversely correlated with CFI (r = 0.67, P = 0.017) whereas IL-6 positively correlated with CFI (r = 0.76, P = 0.004). CONCLUSIONS: Acute myocardial infarction is associated with a significant rise in TNF-alpha and IL-6 levels independent of infarct size or myonecrosis. Tumour necrosis factor-alpha and IL-6 correlate dichotomously with CFI indicating differing roles in reperfused AMI. PMID- 15818084 TI - Plasma homocysteine levels and the left ventricular systolic function in coronary artery disease patients. AB - BACKGROUND: Numerous studies have shown a relationship between hyperhomocysteinemia, atherothrombosis and cardiovascular mortality. However, an association between hyperhomocysteinemia and the extent of coronary artery disease (CAD) remains controversial whereas its relationship with left ventricular systolic function has not been established. METHODS: One hundred and fifty-seven patients with angiographically defined CAD were included. The relationships between hyperhomocysteinemia, severity of CAD and left ventricular systolic function were studied. Left ventricular systolic function was determined primarily by ventriculography. The severity of CAD was determined through coronary angiography using the Gensini score and the number of vessels with > or = 50% stenosis. RESULTS: The mean fasting plasma homocysteine level was 13.4 mumol/l+/-0.5 SE. Elevated levels of homocysteine correlated significantly with increased severity of CAD both by the Gensini scores (r-value = 0.344, P < 0.0005) and the total number of diseased vessels (r-value = 0.387, P < 0.0005). The patients with hyperhomocysteinemia were found to have significantly reduced left ventricular ejection fraction (r-value = -0.382, P < 0.0005). A multivariate regression analysis revealed homocysteine level to be an independent predictor of left ventricular systolic function. In addition, adjusted analysis revealed hyperhomocysteinemia to be associated with global left ventricular dysfunction. CONCLUSION: In patients with CAD, homocysteine levels correlate independently with left ventricular systolic function. The mechanism of this association between homocysteine and left ventricular systolic function is unknown but may be due to a direct effect of homocysteine on myocardial function separate from its effects on coronary atherosclerosis. PMID- 15818085 TI - Correlation between endocardial voltage mapping and myocardial perfusion: implications for the assessment of myocardial ischemia. AB - BACKGROUND: Recent clinical studies using a non-fluoroscopic three-dimensional (3D) left ventricular (LV) mapping system suggested reduced endocardial voltage amplitudes measured in zones sustaining myocardial ischemia or infarction. However, the direct relationship between myocardial perfusion and endomyocardial voltage amplitudes has not been fully elucidated. METHODS: In a pig model of chronic myocardial ischemia (n = 20), LV endocardial unipolar voltage (UpV) mapping was performed using the Biosense 3D navigation system (Johnson and Johnson, Warren, New Jersey, USA) 4 weeks after ameroid constrictor placement around the left circumflex coronary artery. Echocardiography was used to assess regional changes in myocardial wall thickening (MT) and fluorescent microspheres (4 x 10/injection) were used to quantify rest regional myocardial blood flow (MBF) in ischemic (left circumflex) and remote non-ischemic (left anterior descending) regions. RESULTS: UpV measurements were reduced in ischemic compared to non-ischemic zones (9.9+/-3.3 compared with 13.3+/-3.3 mV, P = 0.03). This corresponded to changes in endocardial MBF and MT, which were both noted to be significantly reduced in the ischemic compared to the non-ischemic area (MBF, 0.50+/-0.16 compared with 0.74+/-0.15 ml/g per min, P = 0.001; MT, 26.1+/-12.0 compared with 37.4+/-10.1%, P=0.003). A positive linear correlation was found between UpV at rest and endomyocardial (but not epicardial) perfusion: UpV (mV) = 7.8+5.9xMBFendocardial (r = 0.32, P = 0.05). CONCLUSIONS: Chronic myocardial ischemia, resulting in reduced perfusion and function at rest (that is, hibernating myocardium), is characterized by a significant reduction ( approximately 25%) in endocardial UpV potentials, which correlates with reduced endomyocardial blood flow and tissue perfusion at rest. PMID- 15818086 TI - Long-term follow-up after deferral of coronary intervention based on myocardial fractional flow reserve measurement. AB - OBJECTIVE: To assess long-term results after deferring coronary intervention (percutaneous coronary intervention (PCI)) of an intermediate lesion with a value of myocardial fractional flow reserve (FFR) > or = 0.75 in a 'real life' patient population with no respect to results of stress tests (if performed) or coronary disease extent. METHODS: PCI of an intermediate lesion was deferred in a group of 85 consecutive patients (54 men, 61+/-10 years) on the basis of the result of FFR > or = 0.75 (mean FFR, 0.89+/-0.06%). FFR was measured in 111 stenoses (mean diameter stenosis, 54+/-8%, left anterior descending coronary artery, 65 (58%), left circumflex coronary artery, 24 (22%), right coronary artery, 22 (20%). Multi vessel disease (defined as visually assessed diameter reduction of more than 50% in at least two arteries of more than 1.5 mm diameter, supplying at least two of the three major coronary artery perfusion territories) was present in 67% of patients (one-vessel disease, 28 patients (33%), two-vessel disease, 39 patients (46%), three-vessel disease, 18 patients (21%). Recorded events during follow-up were as follows: all-cause death, cardiac death, non-fatal myocardial infarction, ischemia-driven target lesion transcatheter revascularization (TLR) and coronary artery bypass graft (CABG). Angina class (Canadian Cardiovascular Society (CCS) classification) and the need for anti-anginal drugs were recorded. RESULTS: Follow-up was completed in 85 patients (100%). Mean duration of follow-up was 22.6+/-6.6 months (range 4-33 months). Events occurred in 11 patients (13%). Seven patients died; this included two cardiac deaths. A non-fatal myocardial infarction occurred in one patient, one patient needed TLR and three patients underwent CABG. Estimated 33 month cardiac-event-free survival (Kaplan-Meier) was 91+/-4%. Angina class decreased [1.6+/-1.2 compared with 0.8+/-0.8 (P < 0.0001)] without difference with respect to the use of anti-anginal drugs (1.7+/-0.8 compared with 1.7+/-0.9, P = NS). CONCLUSIONS: Deferring coronary interventions of intermediate stenosis based on FFR measurement is safe with respect to long term follow-up, irrespective of the extent of coronary artery disease. PMID- 15818087 TI - Determinants of endothelial dysfunction and carotid intima-media thickness in combined hyperlipidemia. AB - BACKGROUND: Early functional and morphological changes in the course of the atherosclerotic process are manifested as endothelial dysfunction and increased intima-media thickness (IMT) of the arterial wall. These are both associated with various atherosclerotic risk factors. We investigated whether the same factors are associated with functional and morphological changes of the arterial wall in men with combined hyperlipidemia. METHODS: Flow-mediated dilatation (FMD) of the brachial artery and carotid IMT were measured in 72 male patients aged 46+/-5 years with combined hyperlipidemia. Serum lipoproteins, fibrinolytic and coagulation parameters, blood glucose, proinflammatory cytokines and C-reactive protein were also measured. RESULTS: Tumor necrosis factor-alpha, interleukin 6 and apolipoprotein (apo) B were found to be independent predictors of FMD, explaining 87% of FMD variability in multivariate analysis. On the other hand, total tissue factor pathway inhibitor and apo B were independent predictors of increased carotid IMT, explaining 82% of the variation in carotid IMT. CONCLUSIONS: Apo B, which is a marker for the presence of the atherogenic lipoproteins, is associated with both functional and morphological changes of the artery wall. In addition, in asymptomatic overweight middle-aged men with combined hyperlipidemia, functional changes are associated with proinflammatory cytokines, while morphological changes are associated with coagulation parameters. PMID- 15818088 TI - Risk prediction in patients with chest pain: early assessment by the combination of troponin I results and electrocardiographic findings. AB - OBJECTIVE: To evaluate the prognostic value of point of care troponin I (TnI) results in combination with findings from the admission electrocardiogram (ECG) in patients with chest pain. METHODS: Rapid measurements of TnI were performed in 191 consecutive patients with chest pain and a non-diagnostic ECG for myocardial infarction. RESULTS: Within 6 h from admission, maximum TnI elevations of > or = 0.07 microg/l and > or = 0.1 microg/l were noted in 59 and 39% of all patients, respectively. TnI elevations in the range of 0.07-0.09 microg/l were found in many patients with diagnoses other than acute coronary syndrome. By 6-month follow-up, cardiac death had occurred in 7.1 and 11% of patients with maximum TnI > or = 0.07 microg/l and > or = 0.1 microg/l, respectively and myocardial reinfarction was documented in 12 and 15%, respectively. ST-segment depression on the admission ECG was present in 16% of all patients and was the electrocardiographic abnormality with the highest risk (cardiac death 7.7%, myocardial reinfarction 15%). The combination of TnI > or = 0.1 microg/l and ST segment depression or an abnormal admission ECG in general allowed the identification of patients at low, intermediate and high cardiac risk, 3 h after admission. CONCLUSION: A threshold of TnI > or = 0.1 microg/l corresponding to the 10% coefficient of variation is prognostically most suitable for prediction of cardiac events in patients with chest pain. The combination of TnI results and findings from the admission ECG improves prognostic assessment and allows early and reliable risk stratification in this patient population. PMID- 15818089 TI - Apoptosis and angiogenesis are induced in the unstable coronary atherosclerotic plaque. AB - OBJECTIVE: Apoptosis and angiogenesis may be involved in the pathogenesis of atherosclerosis and plaque destabilization. In this study, we investigated if apoptosis and angiogenesis were induced in the unstable human coronary atherosclerotic plaque compared to stable atherosclerotic plaque. METHODS: Atherosclerotic plaques from patients with stable (n = 9) and unstable angina (n = 13) were obtained by directional coronary atherectomy performed during percutaneous transluminal coronary angioplasty. Apoptosis was detected by terminal deoxynucleotidyl transferase end labelling (TUNEL), as well as by immunostaining for caspase 3, Bax and Bcl-2. Neovascularization was determined by immunostaining for the endothelial cell-specific CD31, vascular endothelial growth factor (VEGF-A), angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), hypoxia inducible factor-1alpha (HIF-alpha), and the sections were quantified blindly. RESULTS: The apoptotic nuclei were more frequently found in the unstable coronary atherosclerotic plaques. When the number of apoptotic cells was quantified, an increased apoptotic index was found in the unstable plaques (P = 0.04). The positive staining for caspase-3 was increased in the unstable plaques (P = 0.0008), while no difference in either Bax or Bcl-2 was found between groups. Neovascularization, as evidenced by lumens surrounded by a CD31 positive endothelial layer, was more frequently present in the plaques from patients with unstable angina (P = 0.04). The number of cells with positive staining for VEGF-A was increased in unstable plaques (P = 0.005). No difference of Ang I, Ang II, HIF1-alpha was found between groups. CONCLUSIONS: In unstable human coronary plaques, apoptosis probably involving caspase 3 was found. The plaques had an increased neovascularization, probably induced by VEGF-A. These factors may contribute to explaining plaque destabilization and intraplaque haemorrhage. PMID- 15818090 TI - Vitamin K1 intake and coronary calcification. AB - OBJECTIVES: The activity of matrix Gla-protein (MGP), a potent inhibitor of vascular calcification, is dependent on carboxylation using vitamin K as a co factor. In animals, low intake of total vitamin K has been shown to accelerate vascular calcification via the MGP mechanism. This has led to the hypothesis that low levels of dietary vitamin K intake may be a risk factor for accelerated vascular calcification in humans due to decreased MGP activity. Additionally, some authors have suggested that current recommended daily intake values for vitamin K might be insufficient to fully inhibit vascular calcification via the MGP mechanism. The aim of this study was to examine the relationship between dietary vitamin K1 (the most prevalent dietary form of vitamin K) intake and premature coronary artery calcification (CAC) in an asymptomatic screening population. METHODS: We conducted a prospective study of 807 consecutive active duty US Army personnel, 39-45 years of age, without known coronary heart disease. Vitamin K1 intake was measured with the Block Dietary Questionnaire and CAC was identified using electron-beam computed tomography (EBCT). RESULTS: We found no significant correlation between CAC score and vitamin K1 intake (r = 0.132, P = 0.106). Multivariate analysis with adjustment for cardiac risk factors showed no association between dietary vitamin K1 intake and CAC. CONCLUSIONS: Dietary vitamin K1 (phylloquinone) intake appears to be unrelated to premature coronary calcification in a screening population. Further investigation into the relationship of vascular calcification and other forms of vitamin K1 (menaquinones) is indicated. PMID- 15818091 TI - Bibliography current world literature. PMID- 15818092 TI - Adverse effect on a referral intensive care unit's performance of accepting patients transferred from another intensive care unit. AB - OBJECTIVE: To determine whether observed and predicted mortality for intensive care unit (ICU) transfer admissions is different from non-ICU transfer admissions and how that might affect ICU performance evaluation. DESIGN, SETTING, AND PATIENTS: We retrospectively analyzed the charts of 3,416 patients admitted to our tertiary referral ICU from January 1995 to December 2001 and evaluated the effect on our performance (based on the Simplified Acute Physiology Score II risk model) of accepting patients transferred from another hospital's ICU. MAIN RESULTS: During the study period, 597 patients (17%) had been transferred from a non-ICU setting in another hospital (hospital transfer) and 408 (12%) from another hospital's ICU (ICU transfer). ICU mortality and standardized mortality ratios were significantly higher for ICU-transfer patients than for hospital transfer or directly admitted patients: 34% vs. 23% vs. 17% (p < .0001) and 0.95 (95% confidence interval, 0.83-1.08), 0.82 (95% confidence interval, 0.71-0.95), and 0.62 (95% confidence interval, 0.55-0.68), respectively. ICU-transfer patients had 3.6-fold longer mean ICU stays and 1.9-fold longer durations of mechanical ventilation than directly admitted patients. Hospital-transfer (odds ratio = 1.89) and ICU-transfer patients (odds ratio = 2.41) had significantly higher mortality rates, even after adjustment for case mix and disease severity. Consequently, a benchmarking program adjusting only for these latter variables, but not admission source, would penalize our ICU by 39 excess deaths per 1,000 admissions as compared with another ICU admitting no transfer patients. Finally, patients transferred from the ward of another hospital had significantly higher mortality rates (odds ratio = 1.56) as compared with patients directly admitted from the ward of our hospital, confirming the "transfer effect" for this homogeneous patients' subgroup. CONCLUSIONS: Admission source remains a strong and independent predictor of ICU death, despite adjustment for case mix and disease severity at ICU admission. Specifically, accepting numerous ICU-transfer patients, for whom the probability of ICU death is the most underestimated by a system adjusting only for case mix and disease severity, can adversely affect the evaluation of referral centers' performance. Future benchmarking and profiling systems should evaluate and adequately account for the ICU-transfer factor to provide healthcare payers and consumers with more accurate and valid information on the true performance of referral centers. PMID- 15818093 TI - Critical illness polyneuropathy and myopathy in patients with acute respiratory distress syndrome. AB - OBJECTIVE: Critical illness polyneuropathy/myopathy (CIP/CIM) is frequently described in critically ill patients who survive severe sepsis. Clinically relevant paresis is major symptom of CIP/CIM. We aimed at determining risk factors and diagnostic value of electrophysiologic testing for CIP/CIM in patients with acute respiratory distress syndrome (ARDS). DESIGN: Single-center, retrospective analysis, using charts. SETTING: University medical center. PATIENTS: Fifty consecutive ARDS patients in our intensive care unit. INTERVENTIONS: Patient characteristics and clinical course were analyzed. All patients received early electrophysiologic testing. CIP/CIM was diagnosed by the presence of clinical relevant paresis. MEASUREMENTS AND MAIN RESULTS: Clinically relevant paresis was confirmed in 27 ARDS patients (60%), whereas in 18 patients no paresis was determined (controls); five patients died before clinical assessment of paresis was feasible. Patients with paresis were older, had more days on mechanical ventilation, and had increased intensive care unit length of stay compared with controls. Patients who developed paresis had elevated daily peak blood glucose levels during 28 days of intensive care unit treatment: 166 (134, 200) mg/dL in CIP/CIM patients vs. 144 (132, 161) mg/dL in controls (median, quartiles). Twenty-five of 27 patients with paresis revealed reduced motor unit potentials, fibrillation potentials, or positive sharp waves on early electrophysiologic testing indicating CIP/CIM, whereas 16 of 18 control patients did not. CONCLUSIONS: In ARDS patients, paresis is a frequent complication causing prolonged mechanical ventilation and intensive care unit length of stay. An association between hyperglycemia and CIP/CIM has been found. However, since this is a retrospective survey, a causal relation is not clearly supported. In this study, the use of early electrophysiologic testing in ARDS patients was a valuable diagnostic tool for detecting CIP/CIM. PMID- 15818094 TI - Immune dysfunction in endotoxicosis: role of nitric oxide produced by inducible nitric oxide synthase. AB - OBJECTIVE: To investigate whether stimulation with lipopolysaccharide (LPS) alters cytokine production by splenocytes in mice and whether it changes the T helper 1 (Th1)/Th2 balance. The role of nitric oxide in such immunologic changes was also explored using mice with genetic lack of inducible nitric oxide synthase (iNOS). DESIGN: : Prospective animal study with concurrent controls. SETTING: University research laboratory. SUBJECTS: iNOS knockout mice and wild-type littermates. INTERVENTIONS: iNOS knockout mice or wild-type mice were injected with LPS or saline with or without anti-interleukin (IL)-6 antibody, and survival was monitored for 7 days. MEASUREMENTS AND MAIN RESULTS: At 24 and 48 hrs after administration, blood samples and splenocytes were obtained to examine immunologic variables. Cell surface markers and cytokine expression of splenocytes were used to characterize the Th1/Th2 balance and were measured by flow cytometry. At 48 hrs after LPS administration, the Th1/Th2 balance shifted toward Th2 predominance in wild-type mice, irrespective of the IL-6 level, whereas it showed Th1 predominance in iNOS knockout mice, and the increase of IL 6 and IL-10 in response to LPS persisted in these animals. After LPS administration, the mortality rate was significantly higher in iNOS knockout mice than in wild-type mice, irrespective of the IL-6 level. CONCLUSIONS: These findings suggest that nitric oxide produced by iNOS during endotoxemia may be involved in down-regulation of Th1 cytokines and up-regulation of Th2 cytokines, whereas IL-6 has no such role. The increased lethality of LPS in iNOS knockout mice suggests that nitric oxide may be protective against proinflammatory cytokine-induced damage. Nitric oxide excess may increase susceptibility to nosocomial infections, so-called immunoparalysis. PMID- 15818095 TI - Transfusion-related acute lung injury: definition and review. AB - BACKGROUND: Transfusion-related acute lung injury (TRALI) is now the leading cause of transfusion-associated mortality, even though it is probably still underdiagnosed and underreported. NATIONAL HEART, LUNG, AND BLOOD INSTITUTE ACTION: The National Heart, Lung, and Blood Institute convened a working group to identify areas of research needed in TRALI. The working group identified the immediate need for a common definition and thus developed the clinical definition in this report. MAJOR CONCEPTS IN THE DEFINITION: The major concept is that TRALI is defined as new acute lung injury occurring during or within 6 hrs after a transfusion, with a clear temporal relationship to the transfusion. Also, another important concept is that acute lung injury temporally associated with multiple transfusions can be TRALI, because each unit of blood or blood component can carry one or more of the possible causative agents: antileukocyte antibody, biologically active substances, and other yet unidentified agents. RECOMMENDATION: Using the definition in this report, clinicians can diagnose and report TRALI cases to the blood bank; importantly, researchers can use this definition to determine incidence, pathophysiology, and strategies to prevent this leading cause of transfusion-associated mortality. PMID- 15818096 TI - The effect of prompt physician visits on intensive care unit mortality and cost. AB - OBJECTIVE: To determine the effect on mortality, length of stay, and direct variable cost of physician response time to seeing patients after intensive care unit admission. DESIGN: Retrospective analysis of the intensive care unit database. SETTING: Medical center. PATIENTS: Subjects were 840 patients who had complete direct variable cost data and a subset of 316 patients who were matched by propensity scores. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Median time to first visit by a physician was 6 hrs. One hundred thirty-five patients (16.1%) died in hospital compared with 25.0% predicted by Acute Physiology and Chronic Health Evaluation risk (p < .001). Higher Acute Physiology and Chronic Health Evaluation risk, older age, mechanical ventilation on arrival in the intensive care unit, and longer time until seen by a physician were predictors of hospital mortality. Each 1-hr delay in seeing the patient was associated with a 1.6% increased risk of hospital death, which further increased to 2.1% after including propensity score. However, patients seen more promptly (<6 vs. >6 hrs) had greater hospital direct variable cost ($11,992 +/- $12,043 vs. $10,355 +/- $10,368, p = .04), before controlling for acuity of illness and other factors that may have affected time to evaluation. In the subpopulation of propensity matched patients, patients seen promptly (<6 vs. >6 hrs) had shorter hospital length of stays (11 +/- 11 vs. 13 +/- 14 days, p = .03) but similar direct variable costs ($10,963 +/- 10,778 vs. $13,016 +/- 13,006, p = .16) and similar mortality rates (24 vs. 30, p = .46). CONCLUSIONS: In the total patient population, delay in seeing patients was associated with an increased risk of death. In the propensity-matched patients, promptly seen patients had shorter hospital stays but similar direct variable costs. PMID- 15818098 TI - Clinical evaluation of an inspiratory impedance threshold device during standard cardiopulmonary resuscitation in patients with out-of-hospital cardiac arrest. AB - OBJECTIVE: To determine whether an impedance threshold device, designed to enhance circulation, would increase acute resuscitation rates for patients in cardiac arrest receiving conventional manual cardiopulmonary resuscitation. DESIGN: Prospective, randomized, double-blind, intention-to-treat. SETTING: Out of-hospital trial conducted in the Milwaukee, WI, emergency medical services system. PATIENTS: Adults in cardiac arrest of presumed cardiac etiology. INTERVENTIONS: On arrival of advanced life support, patients were treated with standard cardiopulmonary resuscitation combined with either an active or a sham impedance threshold device. MEASUREMENTS AND MAIN RESULTS: We measured safety and efficacy of the impedance threshold device; the primary end point was intensive care unit admission. Statistical analyses performed included the chi-square test and multivariate regression analysis. One hundred sixteen patients were treated with a sham impedance threshold device, and 114 patients were treated with an active impedance threshold device. Overall intensive care unit admission rates were 17% with the sham device vs. 25% in the active impedance threshold device (p = .13; odds ratio, 1.64; 95% confidence interval, 0.87, 3.10). Patients in the subgroup presenting with pulseless electrical activity had intensive care unit admission and 24-hr survival rates of 20% and 12% in sham (n = 25) vs. 52% and 30% in active impedance threshold device groups (n = 27) (p = .018, odds ratio, 4.31; 95% confidence interval, 1.28, 14.5, and p = .12, odds ratio, 3.09; 95% confidence interval, 0.74, 13.0, respectively). A post hoc analysis of patients with pulseless electrical activity at any time during the cardiac arrest revealed that intensive care unit and 24-hr survival rates were 20% and 11% in the sham (n = 56) vs. 41% and 27% in the active impedance threshold device groups (n = 49) (p = .018, odds ratio, 2.82; 95% confidence interval, 1.19, 6.67, and p = .037, odds ratio, 3.01; 95% confidence interval, 1.07, 8.96, respectively). There were no statistically significant differences in outcomes for patients presenting in ventricular fibrillation and asystole. Adverse event and complication rates were also similar. CONCLUSIONS: During this first clinical trial of the impedance threshold device during standard cardiopulmonary resuscitation, use of the new device more than doubled short-term survival rates in patients presenting with pulseless electrical activity. A larger clinical trial is underway to determine the potential longer term benefits of the impedance threshold device in cardiac arrest. PMID- 15818099 TI - Low spatial resolution computed tomography underestimates lung overinflation resulting from positive pressure ventilation. AB - OBJECTIVE: In acute lung injury, lung overinflation resulting from mechanical ventilation with positive end-expiratory pressure (PEEP) can be assessed using lung computed tomography. The goal of this study was to compare lung overinflation measured on low and high spatial resolution computed tomography sections. DESIGN: Lung overinflation was measured on thick (10-mm) and thin (1.5 mm) computed tomography sections obtained at zero end-expiratory pressure (ZEEP) and PEEP 10 cm H2O using a software including a color-coding system. SETTING: A 20-bed surgical intensive care unit of a university hospital. PATIENTS: Thirty mechanically ventilated patients with acute lung injury. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Overinflated lung volume was measured as the end expiratory volume of lung regions with computed tomography attenuations <-900 Hounsfield units. Lung overinflation, expressed in percentage of the total lung volume, was significantly underestimated by thick computed tomography sections compared with thin computed tomography sections (0.4 +/- 1.6% vs. 3.0 +/- 4.0% in ZEEP and 1.9 +/- 4% vs. 6.8 +/- 7.3% in PEEP, p < .01). In patients with a diffuse loss of aeration, the overinflated lung volumes of thick and thin computed tomography sections were, respectively, 0.6 +/- 0.8 mL vs. 16 +/- 10 mL in ZEEP (p < .01) and 8 +/- 9 mL vs. 73 +/- 62 mL in PEEP (p < .05). In patients with a focal loss of aeration, this underestimation was more pronounced: 18 +/- 56 mL vs. 127 +/- 140 mL in ZEEP (p < .01) and 85 +/- 161 mL vs. 322 +/- 292 mL in PEEP (p < .01). CONCLUSIONS: In patients with acute lung injury, an accurate computed tomography estimation of lung overinflation resulting from positive pressure mechanical ventilation requires high spatial resolution computed tomography sections, particularly when the lung morphology shows a focal loss of aeration. PMID- 15818100 TI - Predictors of intensive care unit refusal in French intensive care units: a multiple-center study. AB - OBJECTIVE: To identify factors associated with granting or refusing intensive care unit (ICU) admission, to analyze ICU characteristics and triage decisions, and to describe mortality in admitted and refused patients. DESIGN: Observational, prospective, multiple-center study. SETTING: Four university hospitals and seven primary-care hospitals in France. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Age, underlying diseases (McCabe score and Knaus class), dependency, hospital mortality, and ICU characteristics were recorded. The crude ICU refusal rate was 23.8% (137/574), with variations from 7.1% to 63.1%. The reasons for refusal were too well to benefit (76/137, 55.4%), too sick to benefit (51/137, 37.2%), unit too busy (9/137, 6.5%), and refusal by the family (1/137). In logistic regression analyses, two patient-related factors were associated with ICU refusal: dependency (odds ratio [OR], 14.20; 95% confidence interval [CI], 5.27-38.25; p < .0001) and metastatic cancer (OR, 5.82; 95% CI, 2.22-15.28). Other risk factors were organizational, namely, full unit (OR, 3.16; 95% CI, 1.88-5.31), center (OR, 3.81; 95% CI, 2.27-6.39), phone admission (OR, 0.23; 95% CI, 0.14-0.40), and daytime admission (OR, 0.52; 95% CI, 0.32-0.84). The Standardized Mortality Ratio was 1.41 (95% CI, 1.19-1.69) for immediately admitted patients, 1.75 (95% CI, 1.60-1.84) for refused patients, and 1.03 (95% CI, 0.28-1.75) for later-admitted patients. CONCLUSIONS: ICU refusal rates varied greatly across ICUs and were dependent on both patient and organizational factors. Efforts to define ethically optimal ICU admission policies might lead to greater homogeneity in refusal rates, although case-mix variations would be expected to leave an irreducible amount of variation across ICUs. PMID- 15818101 TI - Ventilator-associated pneumonia: Increased bacterial counts in bronchoalveolar lavage by using urea as an endogenous marker of dilution. AB - OBJECTIVE: Invasive diagnostic procedures such as bronchoalveolar lavage (BAL) with quantitative microbiological cultures are currently recommended for the diagnosis of nosocomial pneumonia. Commonly, in clinical practice, a threshold of > or =10 colony forming units/mL is used for therapeutic decisions. The use of these measurements in daily practice assumes that their repeatability is acceptable. However, many variations among the positive results have been noted. One of the most important is dilution of BAL, which may influence the quantitative results by minimizing bacterial counts. Knowledge of the extent of dilution may increase dramatically the value of quantitative cultures. The aim of this study was to determine to what extent specimens are diluted in BAL by measuring urea in BAL and blood. Furthermore, the impact of a potential dilution effect on the diagnosis of ventilator-associated pneumonia was studied. PATIENTS AND SETTING: A total of 47 patients with ventilator-associated pneumonia in two medical intensive care units at the Vienna General Hospital, a university affiliated facility. DESIGN: Prospective study performed between January 2001 and July 2002. METHODS: BAL fluid was divided immediately into two samples: one for direct microscopic examination of cytocentrifuge preparations for Gram staining to determine percentages of cells containing intracellular bacteria and one for quantitative cultures according to the Cumitech 7A guidelines. Epithelial lining fluid volume was calculated using urea as a marker of dilution and correlated with colony forming units per milliliter. RESULTS: Nineteen out of 47 patients (40%) revealed significant bacterial growth (> or =10 colony forming units/mL). Eight additional patients (17%) would have reached the cutoff level after correction of the dilution effect, which varied between 1.8- and 130-fold. CONCLUSIONS: Data suggest a great variation of dilution during BAL procedures, which influences quantitative results. Using urea to determine the dilution quotient could increase the value of bacterial thresholds in the diagnosis and therapeutic decision of ventilator-associated pneumonia. PMID- 15818102 TI - Randomized, double-blind comparison of immediate-release omeprazole oral suspension versus intravenous cimetidine for the prevention of upper gastrointestinal bleeding in critically ill patients. AB - OBJECTIVE: To demonstrate that a new immediate-release omeprazole oral suspension is effective in preventing upper gastrointestinal bleeding in critically ill patients. DESIGN: A noninferiority analysis was used to compare rates of clinically significant upper gastrointestinal bleeding in a prospective, phase 3, double-blind trial with parallel omeprazole suspension and cimetidine treatment groups. SETTING: A total of 47 intensive care units in the United States. PATIENTS: A total of 359 critically ill patients who required mechanical ventilation for > or =48 hrs, had an Acute Physiology and Chronic Health Evaluation score of > or =11 at baseline, had an intact stomach with a nasogastric or orogastric tube in place, and had at least one additional risk factor for upper gastrointestinal bleeding. INTERVENTIONS: Patients were randomized to treatment with omeprazole suspension (two 40-mg doses on day 1, via orogastric or nasogastric tube, and 40 mg each day thereafter) or intravenous cimetidine (300-mg bolus and 50 mg/hr thereafter) for up to 14 days. Gastric aspirates were sampled for bleeding and pH. Medication doses were doubled for failure of pH control (two successive aspirates with pH < or = 4). MEASUREMENTS AND MAIN RESULTS: Clinically significant upper gastrointestinal bleeding (bright red blood not clearing after 5-10 mins of lavage or persistent Gastroccult positive "coffee-grounds" material for 8 hrs on days 1-2 or for 2-4 hrs on days 3 14 and not clearing with > or =100 mL of lavage) was the primary end point of the trial. The rate of clinically significant bleeding in the per-protocol population was 4.5% with omeprazole suspension and 6.8% with cimetidine, meeting the criteria for the noninferiority of omeprazole suspension. Median gastric pH was > or =6 on all trial days with omeprazole suspension treatment and on 50% of days with cimetidine treatment (p < .001, all trial days). In the omeprazole suspension group, median gastric pH was >4 on each trial day in 95% of patients. CONCLUSIONS: Immediate-release omeprazole suspension is effective in preventing upper gastrointestinal bleeding and more effective than intravenous cimetidine in maintaining gastric pH of >4 in critically ill patients. PMID- 15818103 TI - Low tidal volume ventilation does not increase sedation use in patients with acute lung injury. AB - OBJECTIVE: Low tidal volume ventilation is associated with reduced mortality in patients with acute lung injury, yet concerns exist about patient comfort and the levels of sedation and analgesia required during its use. We compared the doses and duration of sedatives and opioid analgesics in patients receiving low vs. traditional tidal volumes at our institution. DESIGN: Secondary analysis of a randomized clinical trial. SETTING: University-affiliated county hospital in Seattle, WA. PATIENTS: Sixty-one patients with acute lung injury enrolled in the National Heart, Lung and Blood Institute's ARDS Network tidal volume trial. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Thirty-three patients were randomized to the lower tidal volume arm (6 mL/kg of predicted body weight) and 28 patients were randomized to the higher tidal volume arm (12 mL/kg of predicted body weight). There were no significant differences in the percentage of study days patients received sedatives, opioids, or neuromuscular blockade. When specific study days were examined, there were no significant differences in the proportion of patients receiving benzodiazepines, propofol, haloperidol, and opioids on days 1, 2, 3, and 7 of mechanical ventilation, nor were there differences in the doses of benzodiazepines and opioids on those days. CONCLUSIONS: At our institution, low tidal volume ventilation was not associated with increased dose or duration of sedatives in patients with acute lung injury. Sedation administration should not be considered a barrier to implementing a lung protective ventilation strategy. PMID- 15818104 TI - Phospholipases A2 and platelet-activating-factor acetylhydrolase in patients with acute respiratory distress syndrome. AB - OBJECTIVE: Phospholipases A2 (PLA2) comprise a family of enzymes probably implicated in the development of acute respiratory distress syndrome (ARDS). The aim was to investigate PLA2 activities and characteristics in bronchoalveolar lavage (BAL) fluid, BAL cells, and plasma from patients with ARDS by a fluorometric method. DESIGN: Prospective, controlled study. SETTING: Fourteen-bed polyvalent intensive care unit in a university hospital. PATIENTS: A total of 31 mechanically ventilated patients, 20 with and 11 without ARDS, were studied. INTERVENTION: BAL was performed by fiberoptic bronchoscopy in mechanically ventilated patients with a controlled mechanical ventilation mode. MEASUREMENTS: PLA2 and platelet-activating-factor acetylhydrolase were determined in BAL fluid, cells, and plasma. For the classification of PLA2-specific inhibitors, Western blot analysis and their biochemical characteristics were used. RESULTS: In ARDS patients, increased PLA2 levels were detected in BAL fluid, BAL cells, and plasma compared with the control patients. PLA2 in BAL fluid was mainly type IIA secretory and cytosolic types. In plasma, type IIA secretory and cytosolic and a Ca-independent PLA2 were found. In BAL cells, a cytosolic form, probably a Ca independent intracellular form, and a low activity of type IIA secretory PLA2 was also observed. Total PLA2 activity correlated inversely with Pao2/Fio2 ratio and positively with the mortality rate. Patients with direct ARDS exhibited higher PLA2 activity compared with patients with indirect ARDS. Platelet-activating factor acetylhydrolase activity was higher in BAL fluid and plasma, but it was lower in BAL cells. CONCLUSION: Ca-dependent, secretory, cytosolic, and Ca independent forms of PLA2 and platelet-activating-factor acetylhydrolase could play important roles in the development or down-regulation of inflammation in ARDS, respectively. PMID- 15818105 TI - Increasing mean arterial pressure in patients with septic shock: effects on oxygen variables and renal function. AB - OBJECTIVE: To measure the effects of increasing mean arterial pressure on oxygen variables and renal function in septic shock. DESIGN: Prospective, open-label, randomized, controlled study. SETTING: Medical-surgical intensive care unit of a tertiary care teaching hospital. PATIENTS: Twenty-eight patients with a diagnosis of septic shock who required fluid resuscitation and pressor agents to increase and maintain mean arterial pressure > or =60 mm Hg. INTERVENTIONS: Patients were treated with fluid and norepinephrine to achieve and maintain a mean arterial pressure of 65 mm Hg. Then they were randomized in two groups: In the first group (control group, n = 14), mean arterial pressure was maintained at 65 mm Hg, and in the second group (n = 14), mean arterial pressure was increased to 85 mm Hg by increasing the dose of norepinephrine. MEASUREMENTS AND MAIN RESULTS: Hemodynamic variables (mean arterial pressure, heart rate, mean pulmonary artery pressure, pulmonary artery occlusion pressure, cardiac index, systemic vascular resistance index, pulmonary vascular resistance index, left and right ventricular stroke indexes), metabolic variables (oxygen delivery, oxygen consumption-calorimetric method, arterial lactate), and renal function variables (urine flow, serum creatinine, creatinine clearance) were measured. After introduction of norepinephrine, similar values of hemodynamic, metabolic, and renal function variables were obtained in both groups. No changes were observed in group 1 during the study period. Increasing mean arterial pressure from 65 to 85 mm Hg with norepinephrine in group 2 resulted in a significant increase in cardiac index from 4.8 (3.8-6.0) to 5.8 (4.3-6.9) L.min.m. Arterial lactate and oxygen consumption did not change. No changes were observed in renal function variables: urine flow, 63 (14-127) and 70 (15-121) mL; serum creatinine, 170 (117-333) and 153 (112-310) mumol.L; and creatinine clearance, 50 (12-77) and 67 (13-89) mL.min.1.73 m. CONCLUSIONS: Increasing mean arterial pressure from 65 to 85 mm Hg with norepinephrine neither affects metabolic variables nor improves renal function. PMID- 15818106 TI - In situ diagnosis of intravascular catheter-related bloodstream infection: a comparison of quantitative culture, differential time to positivity, and endoluminal brushing. AB - OBJECTIVE: To compare the accuracy of three techniques that do not require central venous catheter removal to diagnose catheter-related bloodstream infection. DESIGN: Prospective cohort study of central venous catheters from suspected cases of catheter-related bloodstream infection. SETTING: University teaching hospital. PATIENTS: One hundred and twenty-five central venous catheters from patients with suspected catheter-related bloodstream infection (a raised peripheral white blood cell count, temperature >37 degrees C, and/or local signs of infection at the catheter skin entry site) in intensive care and surgical patients in a large teaching hospital were assessed. INTERVENTIONS: None. MEASUREMENTS: Three techniques were compared: the differential time to positivity of central venous catheter vs. peripheral-blood cultures, quantitative culture of central venous catheter vs. peripheral blood, and the endoluminal brush with peripheral blood culture. MAIN RESULTS: Central venous catheters with a median dwell time of 11 days were examined. There were 36 episodes of catheter-related bloodstream infection, defined as a positive result from at least two of the three tests in the presence of a peripheral blood culture growing the same microorganism and without an identifiable alternative source of sepsis. The sensitivities of the endoluminal brush, quantitative culture, and differential time to positivity techniques were 100%, 89%, and 72%, respectively, with corresponding specificities of 89%, 97%, and 95%. Blood could be directly aspirated from only 231 of 312 (74%) lumens. In the 20 cases of catheter-related bloodstream infection associated with multiple-lumen central venous catheters, endoluminal brushing was positive for one, two, and three lumens in nine (45%), six (30%), and five (25%) cases, respectively. CONCLUSIONS: All three techniques had relatively high sensitivity. However, inability to obtain samples via central venous catheters is a major drawback of the differential time to positivity and quantitative blood culture approaches. Differential time to positivity is simple to perform and has high specificity and therefore could be used as a first line approach, with the endoluminal brush reserved for cases where blood cannot be obtained. All lumens of multiple-lumen central venous catheters must be sampled to ensure maximal sensitivity. PMID- 15818107 TI - Time-course of sTREM (soluble triggering receptor expressed on myeloid cells)-1, procalcitonin, and C-reactive protein plasma concentrations during sepsis. AB - OBJECTIVE: To describe the course of plasma sTREM (soluble triggering receptor expressed on myeloid cells)-1, procalcitonin (PCT), and C-reactive protein (CRP) concentrations during sepsis and their clinical informative value in predicting outcome. DESIGN: Prospective, noninterventional study. SETTING: Medical adult intensive care unit at a university hospital in France. PATIENTS: Sixty-three critically ill patients with sepsis, severe sepsis, or septic shock. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Soluble TREM-1 concentrations were significantly lower at admission in nonsurvivors (n = 21) than in survivors (n = 42) (94 [30-258] vs. 154 [52-435] pg/mL, p = .02), whereas PCT levels were higher among nonsurvivors (19.2 [0.3-179] vs. 2.4 (0-254) pg/mL, p = .001). CRP levels did not differ between the two groups of patients. Plasma PCT and CRP decreased during the 14-day period of study in both survivors and nonsurvivors. Conversely, sTREM-1 plasma concentrations remained stable or even increased in nonsurviving patients and decreased in survivors. An elevated baseline sTREM-1 level was found to be an independent protective factor with an odds of dying of 0.1 (95% confidence interval, 0.1-0.8). CONCLUSION: A progressive decline of plasma sTREM-1 concentration indicates a favorable clinical evolution during the recovery phase of sepsis. In addition, baseline sTREM-1 level may prove useful in predicting outcome of septic patients. PMID- 15818108 TI - Delayed neutralization of interferon-gamma prevents lethality in primate Gram negative bacteremic shock. AB - OBJECTIVE: Whether anticytokine therapies have a place in the treatment of severe sepsis and septic shock remains a question. Although a number of preclinical studies have shown efficacy in primate models of bacteremic shock when administered prophylactically, these same therapies have a significantly diminished effectiveness when administered therapeutically. This study investigated whether delayed administration of a novel anti-human interferon gamma monoclonal antibody could improve outcome and reduce organ injury in a lethal model of Escherichia coli bacteremia, when administered after the onset of shock. DESIGN: Randomized, prospective, double-blinded intervention study. SUBJECTS: Cynomolgus monkeys. INTERVENTIONS: Treatment with a humanized monoclonal antibody directed against human interferon-gamma (INNO 202), administered after the onset of shock, induced by the infusion of live E. coli. MEASUREMENTS AND MAIN RESULTS: Five of the six vehicle-treated monkeys died or were killed within 24-72 hrs after E. coli administration, and all died within 5 days. In contrast, six of the eight animals treated with the anti-interferon gamma survived for 7 days, and three of the eight animals survived 14 days (p = .013 vs. vehicle). Delayed treatment with the anti-interferon-gamma monoclonal antibody did not restore hemodynamics or reduce the amount of crystalloid containing fluid required to resuscitate the animals but did attenuate renal failure (p < .05) and the magnitude of the inflammatory cytokine response (p < .05). CONCLUSIONS: In a primate model of E. coli bacteremic shock, delayed neutralization of interferon-gamma after the onset of shock improved survival and attenuated the pathologic changes associated with the development of organ dysfunction. These findings suggest that interferon-gamma blockade represents a potentially effective mode of late intervention in lethal septic shock. PMID- 15818109 TI - New generation of hemoglobin-based oxygen carriers evaluated for oxygenation of critically ischemic hamster flap tissue. AB - OBJECTIVES: The aim of this study was to investigate and compare the effects of a traditionally formulated, low-viscosity, right-shifted polymerized bovine hemoglobin solution and a highly viscous, left-shifted hemoglobin vesicle solution (HbV-HES) on the oxygenation of critically ischemic peripheral tissue. DESIGN: Randomized, prospective study. SETTING: University laboratory. SUBJECT: A total of 40 male golden Syrian hamsters. INTERVENTIONS: Island flaps were dissected from the back skin of anesthetized hamsters. The flap included a critically ischemic, hypoxic area that was perfused via a collateralized vasculature. One hour after completion of the preparation, the animals received a 33% blood exchange with 6% hydroxyethyl starch 200/0.5 (HES, n = 9), HbV suspended in HES (HbV-HES, n = 8), or polymerized bovine hemoglobin solution (n = 9). MEASUREMENTS AND MAIN RESULTS: Three hours after the blood exchange, microcirculatory blood flow (laser-Doppler flowmetry) was increased to 262% of baseline for HbV-HES (p < .01) and 197% for polymerized bovine hemoglobin solution (p < .05 vs. baseline and HbV-HES). Partial tissue oxygen tension (bare fiber probes) was only improved after HbV-HES (9.4 torr to 14.2 torr, p < .01 vs. baseline and other groups). The tissue lactate/pyruvate ratio (microdialysis) was elevated to 51 in the untreated control animals, and to 34 +/- 8 after HbV-HES (p < .05 vs. control) and 38 +/- 11 after polymerized bovine hemoglobin solution (not significant). CONCLUSIONS: Our study suggests that in critically ischemic and hypoxic collateralized peripheral tissue, oxygenation may be improved by normovolemic hemodilution with HbV-HES. We attributed this improvement to a better restoration of the microcirculation and oxygen delivery due to the formulation of the solution. PMID- 15818110 TI - Resuscitation from experimental heatstroke by hyperbaric oxygen therapy. AB - OBJECTIVE: Heatstroke is characterized by hyperthermia, vasoplegic shock, and cerebral ischemia and hypoxia. Hyperbaric oxygen (HBO) has been shown to reduce brain ischemia and behavioral dysfunction during cerebral artery occlusion. The efficacy of HBO therapy for resuscitation from heatstroke remains to be determined in the laboratory. DESIGN: Anesthetized rats were randomized to several groups and administered: 1) no resuscitation (normobaric air) after onset of heatstroke, 2) HBO for 1 hr (100% oxygen at 253 kPa for 1 hr), 3) cyclic HBO intermitted by a 5-min air break for 1 hr of treatment (100% oxygen at 253 kPa), 4) hyperbaric air (air at 253 kPa for 1 hr), 5) normobaric hyperoxia (100% oxygen at 101 kPa for 1 hr), or 6) 8% HBO (hyperbaric 8% oxygen at 253 kPa for 1 hr). SETTING: Laboratory investigation. SUBJECTS: Sprague-Dawley rats (300- to 400-g males). INTERVENTIONS: Rats were exposed to an ambient temperature of 43 degrees C to induce heatstroke. Their colonic temperature; mean arterial pressure; heart rate; arterial blood levels of pH, Paco2, Pao2, So2%, and tumor necrosis factor alpha; the cortical levels of ischemic and damage markers, and cortical neuronal damage scores were determined. The moment at which mean arterial pressure began to decrease from peak levels was arbitrarily taken as the onset of heatstroke. MAIN RESULTS: Survival time (interval between onset of heatstroke and animal death) was 19 +/- 1 (n = 10), 131 +/- 18 (n = 14), 159 +/- 28 (n = 13), 72 +/- 14 (n = 10), 68 +/- 12 (n = 10), and 45 +/- 11 (n = 10) mins, respectively, for normobaric air, HBO for 1 hr, cyclic HBO, hyperbaric air, normobaric hyperoxia, and 8% HBO groups. The heatstroke induced arterial hypotension and bradycardia, decreased arterial levels of pH, Pao2, and So2%, increased arterial levels of tumor necrosis factor-alpha, and increased values of cellular ischemia and damage markers. In addition, neuronal damage scores in the cortex were significantly reduced by HBO for 1 hr and cyclic HBO resuscitation. CONCLUSION: We successfully demonstrated that HBO and, to some extent, hyperbaric air, normobaric hyperoxia, or HBO 8% was found beneficial in resuscitating rats with experimental heatstroke. HBO effectively reduced heatstroke-induced arterial hypotension, hypoxia, plasma tumor necrosis factor-alpha overproduction, and cerebral ischemia and damage and improved survival. PMID- 15818111 TI - Adrenomedullin reduces Staphylococcus aureus alpha-toxin-induced rat ileum microcirculatory damage. AB - OBJECTIVE: Increased microvascular permeability and perfusion mismatch are hallmarks of sepsis or septic shock. The intestinal mucosa is very sensitive to tissue hypoxia. Intestinal mucosa dysfunction may allow translocation of bacteria and their products, thereby perpetuating sepsis and inflammation. Staphylococcus aureus alpha-toxin is a major pathogenicity determinant of this bacterium, provoking cardiovascular collapse. Current evidence suggests that the endogenous peptide adrenomedullin stabilizes circulatory homeostasis in systemic inflammatory response. Using alpha-toxin as a well-defined strong initiator of an inflammatory reaction, we tested the hypothesis that exogenously applied adrenomedullin stabilizes gut microcirculation. DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university hospital. SUBJECTS: Isolated, perfused ileum from male Sprague-Dawley rats and human umbilical vein endothelial cells. INTERVENTIONS: Administration of S. aureus alpha-toxin before or after infusion of adrenomedullin. MEASUREMENTS AND MAIN RESULTS: Injection of a bolus of 1 microg of alpha-toxin in the superior mesenteric artery in a constant-flow, blood-perfused preparation of rat ileum increased perfusion pressure and relative hemoglobin concentration and decreased mucosal hemoglobin oxygen saturation. Continuous infusion of adrenomedullin (0.1 micromol/L) significantly reduced these alpha-toxin-related effects. Severe microvascular hyperpermeability observed in alpha-toxin-exposed ileum was abolished by adrenomedullin pretreatment. In addition, adrenomedullin blocked alpha-toxin-induced endothelial myosin light chain phosphorylation, endothelial cell contraction, and subsequent loss of endothelial barrier function in vitro. Treatment of alpha-toxin (infusion of 0.05 microg/mL)-exposed ileum with adrenomedullin (0.1 micromol/L) started 10 mins after onset of toxin application also significantly reduced superior mesenteric artery pressure and permeability increase. CONCLUSIONS: In summary, these data suggest that exogenous adrenomedullin protects ileum by reducing alpha toxin-induced microcirculatory disturbances and by stabilizing endothelial barrier function. PMID- 15818112 TI - Apoptosis inhibition during preservation by fructose-1,6-diphosphate and theophylline in rat intestinal transplantation. AB - OBJECTIVE: This study evaluated the effect of fructose-1,6-diphosphate (FDP), theophylline, or the addition of both together to the preservation solution (University of Wisconsin [UW]) on apoptosis during preservation and the effect of apoptosis minimization on the early reperfusion period after transplantation. DESIGN: Prospective, randomized, and controlled animal study. SETTING: Laboratory of a research institute. SUBJECT: Male Wistar rats. INTERVENTIONS: The jejunum was isolated and preserved for 6 hrs in UW solution. FDP and theophylline were added to the UW solution to evaluate their effects on apoptosis both alone and together. The role of adenosine with respect to FDP was examined by increasing endogenous adenosine. In addition, rats were subjected to intestinal transplantation for the evaluation of the effect of apoptosis on bacterial translocation, histology, and neutrophil infiltration after reperfusion. MEASUREMENTS AND MAIN RESULTS: Caspase-3 activity, assayed both in vitro or by cleaved caspase-3 levels in Western blots or immunohistochemically, and the number of terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick-end labeling (TUNEL)-positive cells decreased with FDP and with theophylline addition to UW solution. Increase of endogenous adenosine reversed the antiapoptotic effect of FDP. FDP and theophylline together demonstrated a more pronounced antiapoptotic effect and prevented bacterial translocation after transplantation. CONCLUSION: Supplementary FDP to UW solution decreased apoptosis through an adenosine-independent mechanism. Addition of theophylline to UW solution decreased both apoptosis and bacterial translocation. Concomitant theophylline and FDP addition to preservation solution is recommended to maintain low levels of apoptosis during intestinal hypothermic preservation and to decrease bacterial translocation. PMID- 15818113 TI - Protective effects of low respiratory frequency in experimental ventilator associated lung injury. AB - OBJECTIVE: To determine whether ventilator-associated lung hyperinflation injury can be attenuated by a reduction in respiratory frequency. DESIGN: Prospective comparative laboratory investigation. SETTING: University medical center research laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Eight groups of isolated, perfused rat lungs were exposed to cyclic ventilation at different respiratory frequencies and tidal volumes. Each group of six to eight lung preparations was assigned to one of four respiratory frequencies (10, 20, 40, or 80 breaths/min) and one of two tidal volumes (5 or 20 mL.kg). Measurement of capillary filtration coefficient (Kf,c), a sensitive index of lung microvascular permeability and injury, was made at baseline and at 30, 60, and 90 mins of the experimental conditions. MEASUREMENTS AND MAIN RESULTS: Lungs exposed to 5 mL.kg tidal volume had no elevation in Kf,c at any time point regardless of respiratory frequency. Lungs exposed to 20 mL. kg tidal volume and a respiratory frequency of 80 had significant elevations in Kf,c at all times after baseline compared with lungs exposed to respiratory frequencies of 10, 20, or 40 (0.14 +/- 0.03, 0.16 +/ 0.02, 0.31 +/- 0.05 vs. 0.76 +/- 0.16). Furthermore, the Kf,c at 90 mins was significantly higher than permeability at baseline in this group (1.53 +/- 0.45 vs. 0.12 +/- 0.02 mL.min.cm H2O.100 g of lung tissue). CONCLUSIONS: Reduction in respiratory frequency to values much lower than normal ameliorated experimental ventilator-induced hyperinflation lung injury as determined by pulmonary capillary filtration coefficient. PMID- 15818114 TI - Effects of hyperbaric treatment in cerebral air embolism on intracranial pressure, brain oxygenation, and brain glucose metabolism in the pig. AB - OBJECTIVE: To evaluate the effects of hyperbaric oxygen treatment after cerebral air embolism on intracranial pressure, brain oxygenation, brain glucose/lactate metabolism, and electroencephalograph. DESIGN: Prospective animal study. SETTING: Hyperbaric chamber. SUBJECTS: Eleven Landrace/Yorkshire pigs. INTERVENTIONS: In 11 anesthetized pigs, intracranial pressure and brain oxygenation were measured with microsensor technology, brain glucose/lactate by microdialysis, and electroencephalograph by conventional methods. After injection of air into the internal carotid artery, animals were treated immediately (at 3 mins; t = 3) or at 60 mins (t = 60) with U.S. Navy Treatment Table 6 for 4.48 hrs. RESULTS: At the end of hyperbaric oxygen treatment, intracranial pressure in the t = 60 group (39 +/- 8 mm Hg) was significantly higher than in the t = 3 group (27 +/- 6 mm Hg), brain oxygenation values for group t = 3 and t = 60 were 66 +/- 14 and 52 +/ 15 mm Hg, respectively (no significant difference from baseline), and there were no pathologic scores in the visually assessed electroencephalograph. However, there was a significant decrease in brain glucose and a significant increase in brain lactate in both groups at the end of the 5-hr study period. CONCLUSIONS: Hyperbaric oxygen treatment initiated at both 3 and 60 mins after embolization decreased the deleterious effects of cerebral air embolism on intracranial pressure and brain metabolism. Therefore, this model appears suitable to test the application of hyperbaric oxygen treatment with a delay >60 mins after embolization, as is often the case in the clinical situation. PMID- 15818115 TI - Role of sensory neuron in reduction of endotoxin-induced hypotension in rats. AB - OBJECTIVE: We attempted to determine whether activation of the sensory neuron contributes to reduction of endotoxin-induced hypotension by inhibiting tumor necrosis factor (TNF)-alpha production via calcitonin gene-related peptide (CGRP) release in rats. DESIGN: Prospective, randomized, controlled study. SETTING: Research laboratory at a university medical center. SUBJECTS: Wistar rats weighing 220-280 g. INTERVENTIONS: Mean arterial blood pressure was measured in rats administered endotoxin intravenously. Animals were pretreated with capsazepine (a vanilloid receptor antagonist), CGRP(8-37) (a CGRP receptor antagonist), and indomethacin before endotoxin administration. Levels of CGRP, 6 keto-prostaglandin F1alpha, TNF-alpha, and cytokine-induced neutrophil chemoattractant (CINC) were measured by enzyme immunoassay methods. The concentration of NO2/NO3 was measured using the Griess reagent. Tissue levels of messenger RNA of the inducible form of nitric oxide synthase (iNOS) and TNF-alpha were determined by reverse transcription polymerase chain reaction. MEASUREMENTS AND MAIN RESULTS: Both lung levels of CGRP and plasma levels of 6-keto prostaglandin F1alpha were increased after intravenous administration of endotoxin (5 mg/kg), peaking at 90 mins after endotoxin administration. Increases in plasma levels of 6-keto-prostaglandin F1alpha at 90 mins after endotoxin administration (766 +/- 134 pg/mL) were inhibited by pretreatment with capsazepine (373 +/- 44 pg/mL, p < .05), CGRP(8-37) (406 +/- 64 pg/mL, p < .05), and indomethacin (154 +/- 40 pg/mL, p < .05). Although none of the pretreatments affected a series of endotoxin-induced responses, including increases in lung tissue levels of TNF-alpha, CINC, and iNOS and the resultant hypotension in animals given 5 mg/kg endotoxin, such pretreatments enhanced these pathologic responses in animals given a smaller dose of endotoxin (1 mg/kg) to the same extent as those induced by 5 mg/kg of endotoxin, suggesting that shock responses induced by 5 mg/kg endotoxin are maximum responses and activation of sensory neurons in endotoxin-treated rats is essentially a reparative response. CONCLUSION: Activation of sensory neurons might contribute to reduction of endotoxin-induced hypotension by releasing CGRP, which is capable of promoting endothelial production of prostacyclin. PMID- 15818116 TI - Absolute and relative adrenal insufficiency in children with septic shock. AB - OBJECTIVE: Corticosteroid replacement improves outcome in adults with relative adrenal insufficiency and catecholamine-resistant septic shock. We evaluated the relationship of absolute and relative adrenal insufficiency to catecholamine resistant septic shock in children. DESIGN: Prospective cohort study. SETTING: University hospital pediatric intensive care unit in Brazil. PATIENTS: Fifty seven children with septic shock. Children with HIV infection, those with a history of adrenal insufficiency, and those submitted to any steroid therapy or etomidate within the week before diagnosis of septic shock were excluded. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A short corticotropin test (250 microg) was performed, and cortisol levels were measured at baseline and 30 and 60 mins posttest. Adrenal insufficiency was defined by a response < or =9 microg/dL. Absolute adrenal insufficiency was further defined by a baseline cortisol <20 microg/dL and relative adrenal insufficiency by a baseline cortisol >20 microg/dL. Absolute adrenal insufficiency was observed in 18% of children, all of whom had catecholamine-resistant shock. Relative adrenal insufficiency was observed in 26% of children, of whom 80% had catecholamine-resistant and 20% had dopamine/dobutamine-responsive shock. All children with fluid-responsive shock had a cortisol response >9 microg/dL. Children with adrenal insufficiency had an increased risk of catecholamine-resistant shock (relative risk, 1.88; 95% confidence interval, 1.26-2.79). However, mortality was independently predicted by chronic illness or multiple organ failure (p < .05), not adrenal insufficiency. CONCLUSIONS: Absolute and relative adrenal insufficiency is common in children with catecholamine-resistant shock and absent in children with fluid responsive shock. Studies are warranted to determine whether corticosteroid therapy has a survival benefit in children with relative adrenal insufficiency and catecholamine-resistant septic shock. PMID- 15818117 TI - Role of vascular endothelial growth factor receptor 1 and vascular endothelial growth factor receptor 2 in the vasodilator response to vascular endothelial growth factor in the neonatal piglet lung. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF) regulates vascular proliferation and causes vasodilation. In the pulmonary circulation, the vasorelaxing effect of VEGF has been attributed to nitric oxide, whereas in other vascular beds, prostacyclin and other mechanisms are also involved. This vascular effect follows binding to two receptors, VEGF receptor 1 (VEGFR1) and VEGF receptor 2 (VEGFR2), the latter of which is thought to be the main receptor responsible for the vasorelaxing effect of VEGF. The role of VEGFR1 in the neonatal pulmonary vasculature remains to be determined. DESIGN: Prospective randomized laboratory investigation. SETTING: Animal laboratory. SUBJECTS: Newborn Yorkshire-Landrace piglets. INTERVENTIONS: To determine the mechanisms of action of VEGF in the neonatal pulmonary vasculature, the effect of VEGF (10-10 M) was tested in isolated perfused piglet lungs, alone and in the presence of a VEGFR2 kinase inhibitor, N-nitro-l-arginine (L-NNA), indomethacin (Indo), L-NNA + Indo, and GF109203X, a protein kinase C inhibitor. The effect of a VEGFR1 agonist, placenta growth factor (PlGF), was also studied with or without L-NNA. Perfusate was collected, and cyclic guanosine monophosphate (cGMP), as well as 6 keto prostaglandin F1alpha and thromboxane B2, the stable metabolites of prostacyclin and thromboxane, respectively, was measured. MEASUREMENTS AND MAIN RESULTS: VEGF caused vasorelaxation with a concomitant increase in cGMP. PlGF also decreased vascular tone and increased cGMP. VEGFR2 kinase inhibitor did not prevent the reduction in perfusion pressure seen with VEGF but blocked the increase in cGMP. Pretreatment with L-NNA completely inhibited VEGF and PlGF vasodilation and prevented the increase in cGMP seen with both agonists. Pretreatment with Indo or GF109203X did not reduce the dilator response to VEGF. CONCLUSIONS: VEGF vasodilation may follow nitric oxide release in the piglet pulmonary circulation. VEGF vasorelaxation may not only occur through binding to VEGFR2, since PlGF, the specific VEGFR1 agonist, also causes vasodilation. Therefore, vasodilator response to VEGF may involve both types of receptor in the neonatal piglet pulmonary vasculature. PMID- 15818118 TI - Recommendations for informed consent forms for critical care clinical trials. AB - BACKGROUND: Many subjects enrolled in research studies have a limited understanding of the research to which they consented. OBJECTIVE: To develop recommendations to enhance comprehensiveness and understanding of informed consent forms used in critical care clinical trials. DESIGN: Consensus process. RECOMMENDATIONS: We provide recommendations regarding the U.S. federally required basic and additional elements of informed consent as applied to critical care clinical trials. We also identify issues that investigators need to address, if relevant, in the informed consent forms of critical care clinical trials. These include the description, in understandable language, of complex and detailed experimental protocols that are the focus of the clinical trial, disclosure of death as a risk factor if mortality is an outcome variable, and the identification of who can legally serve as the prospective subject's surrogate. We also offer suggestions to enhance subjects' understanding of informed consent forms. CONCLUSIONS: The literature on informed consent forms suggest that shorter informed consent forms written at a lower reading level, when read carefully, might provide better subject understanding. Prospective evaluation is needed to determine whether our recommendations enhance the informed consent process. PMID- 15818119 TI - Efficacy and safety of recombinant factor VIIa for treatment of severe bleeding: a systematic review. AB - BACKGROUND: Recombinant activated factor VII (factor VIIa) is a prohemostatic agent that can be used for patients with complicated coagulation disorders. Recombinant factor VIIa is, however, increasingly used for several other indications, including patients with a preexistent normal coagulation system but who experience serious bleeding, for example, after major surgery or trauma. DATA SOURCE: We performed a systematic review of all published and unpublished clinical studies using MEDLINE (1966-2004) and all other sources available to assess the available evidence on the efficacy and safety of recombinant factor VIIa in patients with or without coagulation disorders. STUDY SELECTION: We found 483 articles related to the pharmacologic use of recombinant factor VIIa, including 28 clinical trials, 124 case series, and 176 case reports, which were all considered for this review. DATA SYNTHESIS: Recombinant factor VIIa is an effective and relatively potent prohemostatic agent in approximately 90% of patients with hemophilia and inhibiting antibodies and other types of complex coagulation disorders. The application of recombinant factor VIIa in other patients who experience severe bleeding is promising, and although sound evidence from controlled clinical trials is only scarcely available so far, forthcoming trials are likely to provide more substantiation for this use. Recombinant factor VIIa appears to be relatively safe with a 1-2% incidence of thrombotic complications based on published trials. CONCLUSIONS: More randomized controlled clinical trials are required to assess the efficacy and safety of recombinant factor VIIa for patients without a preexistent coagulation disorder and with severe bleeding. In the meantime, off-label use of recombinant factor VIIa may be considered in patients with life-threatening bleeding. PMID- 15818120 TI - Case of fulminant hepatic failure due to unrecognized peripartum cardiomyopathy. AB - OBJECTIVE: To describe a postpartum patient who presented with fulminant hepatic failure and hepatic coma as a result of unrecognized peripartum cardiomyopathy. DESIGN: Case report. SETTING: Medical intensive care unit of a tertiary care academic medical center. PATIENT: A 35-yr-old woman 5 wks postpartum from an uneventful spontaneous vaginal delivery who was transferred to our institution with fulminant hepatic failure and worsening hepatic encephalopathy of unknown etiology for consideration of liver transplantation. INTERVENTIONS: An echocardiogram was obtained as part of an evaluation for refractory shock and the patient was found to have severe global hypokinesis with an ejection fraction of approximately 15%. She was diagnosed with peripartum cardiomyopathy and treatment with digoxin and afterload reduction was initiated. MEASUREMENTS AND MAIN RESULTS: After initiation of appropriate treatment for dilated cardiomyopathy, the patient's hepatic failure resolved and she made a full recovery. CONCLUSIONS: Congestive heart failure is one of the few treatable causes of fulminant hepatic failure. Congestive heart failure must always be included in the differential diagnosis of fulminant hepatic failure of unknown pathogenesis. PMID- 15818121 TI - Scoring systems: is a fix needed? PMID- 15818122 TI - Critical illness polyneuropathy and myopathy in acute respiratory distress syndrome: more common than we realize! PMID- 15818123 TI - Does nitric oxide control the counter-inflammatory response in endotoxic shock? PMID- 15818124 TI - The quest to improve cardiac arrest survival: Overcoming the hemodynamic effects of ventilation. PMID- 15818125 TI - Acute lung injury: new insights from computed tomography. PMID- 15818126 TI - Still a black box: what do we really know about the intensive care unit admission process and its consequences? PMID- 15818127 TI - Another excuse bites the dust: low tidal volume ventilation does not increase sedation use. PMID- 15818128 TI - Phospholipase A2 and acute lung injury: it's just not that simple. PMID- 15818129 TI - Arterial pressure, vasopressors and septic shock: higher is not necessarily better. PMID- 15818130 TI - Ventilator-induced lung injury: do dynamic factors also play a role? PMID- 15818131 TI - Bubbles in the brain: what to do for arterial gas embolism? PMID- 15818132 TI - One step forward: an advance in understanding of adrenal insufficiency in the pediatric critically ill. PMID- 15818133 TI - Is clinical research and ethics a zero-sum game? PMID- 15818134 TI - Human albumin meta-analysis. PMID- 15818135 TI - Conclusions are misleading. PMID- 15818137 TI - Morbidity in hospitalized patients receiving human albumin: a meta-analysis of randomized, controlled trials. PMID- 15818138 TI - Single-beat evaluation of right ventricular contractility. PMID- 15818140 TI - Decreased mortality with the use of the pulmonary artery catheter? PMID- 15818142 TI - Tobramycin in the medical intensive care unit. PMID- 15818145 TI - Hepatobiliary pathology. AB - PURPOSE OF REVIEW: This review highlights recent publications on hepatobiliary pathology concerning several unusual types of hepatitis, fatty liver disease, disorders of the biliary tree and other topics that have a substantial impact on liver biopsy interpretation. RECENT FINDINGS: In the outbreak of severe acute respiratory syndrome (SARS), many patients had abnormalities in liver function tests. Liver biopsy findings in three cases were reported that showed a generic picture of hepatitis, with exceptionally increased mitotic activity. The role of portal myofibroblasts in cirrhosis was examined in several studies. A newly described lesion, isolated ductular hyperplasia (IDH) was found in patients with prolonged abnormalities of liver function tests of uncertain origin. Hyperplastic, well-differentiated bile ductules were seen on liver biopsy in the absence of any identifiable biliary disease. Hereditary hemochromatosis is now a complex entity with various clinicopathological forms based on mutations in the HFE gene and other iron-homeostatic genes such as transferrin receptor 2 and ferroportin 1. In some of these heritable forms of primary iron overload, stainable iron is present in both hepatocytes and Kupffer cells. After liver transplantation, differentiating recurrent HCV infection from acute rejection on liver biopsy is problematic, with exceptionally low inter- and intra-observer reliability shown in one study. SUMMARY: The hepatitis associated with the SARS coronavirus, Isolated Ductular Hyperplasia in patients with liver function test abnormalities and other topics with pathologic relevance are reviewed. PMID- 15818146 TI - Cholestasis and cholestatic syndromes. AB - PURPOSE OF REVIEW: This review highlights recent advances in understanding the regulation of bile acid transport in cholestasis and the pathogenesis and treatment of a variety of cholestatic conditions. RECENT FINDINGS: Highlights include new understanding of the role of Mrp4 in bile acid homeostasis in cholestasis, new insights into the pathogenesis of specific cholestatic syndromes including primary biliary cirrhosis, primary sclerosing cholangitis, biliary atresia, and progressive familial intrahepatic cholestasis, and clinical trials of therapies for primary biliary cirrhosis, primary sclerosing cholangitis and intrahepatic cholestasis. SUMMARY: Our understanding of the molecular mechanisms of cholestasis is advancing. These advances will hopefully lead to more effective therapies for specific cholestatic conditions. PMID- 15818147 TI - Inherited metabolic liver disease. AB - PURPOSE OF REVIEW: The purpose of this review is to identify and discuss recent findings related to inherited metabolic disorders of the liver that increase our understanding of the pathophysiology and treatment for hemochromatosis and other iron overload disorders, Wilson disease and alpha one antitrypsin deficiency. RECENT FINDINGS: The main theme in the recent discoveries for both iron overload disorders and Wilson disease is our increasing understanding that the phenotypic expression of these disorders are greatly influenced by genes involved in the metabolic pathways for these metals, or influence the progression of liver disease independent of metal metabolism. For example, the role of hepcidin dysregulation in hemochromatosis has been a surprising discovery that provides some mechanistic understanding for the increased iron absorption that is present in this disorder. SUMMARY: Given the recent explosion of information on iron and copper metabolism and the cellular processing of alpha one antitrypsin, the highlights reviewed in this article will help the reader keep up to date with the current understanding of these diseases and potential future approaches to their treatment. PMID- 15818148 TI - Drug-induced liver disease 2004. AB - PURPOSE OF REVIEW: To summarize the salient reviews, studies and case reports and series that dealt with clinical, pathological, methodological, and epidemiological descriptions of drug-induced liver disease in the calendar year 2004. RECENT FINDINGS: While no new causes of drug-induced liver injury were reported for 2004, several new reports of previously recognized hepatotoxins, including herbal products, were published. These include the antiretroviral drugs for HIV and agents to manage tuberculosis. Acetaminophen (APAP) retained its preeminent position as the leading cause of drug-induced acute liver failure, currently accounting for nearly 50% of cases according to the latest figures from the U.S. Acute Liver Failure Study Group. Not surprisingly, APAP also heads the list of drugs and toxins leading to liver transplantation for acute hepatic failure. Efforts to reduce the number of cases of intentional APAP poisonings by restricting the number of tablets sold at any one time in the UK are ongoing, but the success of the program may be lessening, as was pointed out this year. The use of potentially hepatotoxic medications in patients with underlying liver disease was examined with the statins, and they emerged as a safe class for use in this setting. SUMMARY: Given the apparent increasing incidence of acute liver failure attributable to APAP in the US, additional efforts are still needed to better define the risks associated with its use and to further reduce the incidence of severe liver injury from this widely used agent. PMID- 15818149 TI - Autoimmune liver disease. AB - PURPOSE OF REVIEW: This review aims to demonstrate how recent insights into disease behavior and mechanisms and the availability of new drugs can be assimilated into evolving concepts of diagnosis and treatment. RECENT FINDINGS: Autoimmune hepatitis has a global distribution, and its clinical manifestations are similar in different regions. Concurrent immune diseases are common, and immunoglobulin A nephropathy may explain ascites in some patients. Subclinical celiac disease can cause cryptic liver dysfunction or be associated with autoimmune hepatitis. A fulminant presentation that is associated with de novo rather than exacerbated pre-existent disease is possible, and these patients may have centrilobular zone 3 necrosis. Bile duct injury as a background histologic finding should not change the diagnosis or therapy, and the clinical significance of autoantibodies can be determined only by examination of liver biopsy tissue. Molecular mimicry may be important in breaking self-tolerance, and a murine model based on DNA immunization with self-antigens supports this hypothesis. Corticosteroid therapy reduces or prevents hepatic fibrosis, and noninvasive techniques promise to facilitate the development of treatments that enhance this effect. Mycophenolate mofetil is a possible salvage therapy that requires clinical trial, and liver transplantation has a 5-year patient survival of 78% but is frequently followed by recurrent disease and acute rejection. SUMMARY: Autoimmune hepatitis should be considered in all patients with acute or chronic liver disease. Therapy can reduce or prevent fibrosis. The new immunosuppressive drugs should undergo clinical trial before empiric use. Animal models promise to identify key pathogenic sites that can be targeted by therapies. PMID- 15818150 TI - Update on viral hepatitis in 2004. AB - PURPOSE OF REVIEW: This article highlights recent advances in viral hepatitis published from December 2003 to November 2004. Studies reporting novel and clinically relevant findings were selected after a PubMed search. The aim is to provide an up-to-date summary of important developments in viral hepatitis. RECENT FINDINGS: Lamivudine was shown to reduce the rate of long-term complications of hepatitis B virus-induced cirrhosis. Adefovir was effective in suppressing lamivudine-resistant hepatitis B virus. Pegylated interferon alone was as effective as pegylated interferon plus lamivudine in the management of HBeAg-negative chronic hepatitis B. A 24-week course of pegylated interferon plus low-dose ribavirin was optimal in patients with hepatitis C virus infected with genotype 2 or 3, but a 48-week course and standard dose of ribavirin were needed in patients with genotype 1. Pegylated interferon plus ribavirin was fairly well tolerated in HIV-hepatitis C virus coinfected patients with stable HIV infection and resulted in response rates that were only slightly lower than that in patients with hepatitis C virus infection only. A dramatic reduction in hepatitis C virus RNA level was observed after 2 days of treatment with an hepatitis C virus protease inhibitor. SUMMARY: The optimal management of chronic viral hepatitis is evolving rapidly. Newer treatment options for hepatitis B, including pegylated interferon, tenofovir, and combination regimens were shown to be effective in treatment-naive and treatment-experienced patients. In addition, impressive gains were made in the treatment of hepatitis C virus infection in difficult-to-treat patients, including African Americans and those with HIV hepatitis C virus coinfection. PMID- 15818151 TI - Hepatocellular carcinoma. AB - PURPOSE OF REVIEW: The incidence of hepatocellular carcinoma (HCC) is expected to continue to increase over the next two decades. The risk factors for the development of HCC are unknown and there is a lack of standardization of the diagnostic criteria for HCC. Our aim is to review the latest information regarding the risk factors, surveillance and diagnosis of this tumor. RECENT FINDINGS: Alcohol, tobacco, obesity, diabetes and viral etiology interact together to increase the risk of hepatocellular carcinoma in patients with cirrhosis, which may allow us to identify a high-risk group for HCC among patients with cirrhosis. Several studies showed that surveillance of cirrhotic patients is cost-effective and leads to an overall improvement in survival. This year a study from the United Network of Organ Sharing in the United States indicated that 30% of patients were understaged by imaging pre-transplant and 31% of patients with a diagnosis of stage 1 HCC (single lesion < 2 cm in diameter) did not have a tumor on the explant examination. Another study showed that washout of arterially enhancing lesions is very sensitive and specific for a diagnosis of HCC. SUMMARY: Not all patients with cirrhosis have an equal risk for developing hepatocellular carcinoma. Therefore, further studies should stratify the risk of HCC so surveillance is tailored to those at the highest risk. Standardization of the diagnostic criteria of HCC is critically important for better patient care and future research. Washout of arterially enhancing lesions should be important criteria of HCC. PMID- 15818152 TI - Portal hypertension. AB - PURPOSE OF REVIEW: Report on significant advances in the pathophysiology, diagnosis, and management of the complications of portal hypertension that have occurred in the last year. RECENT FINDINGS: The specific areas reviewed refer to experimental studies aimed at modifying the factors that lead to portal hypertension (increased intrahepatic vascular resistance and splanchnic vasodilatation) and recent advances in the diagnosis and management of the complications of portal hypertension. The specific complications reviewed in this paper are varices and variceal bleeding (primary prophylaxis, treatment of the acute episode, and secondary prophylaxis), ascites and some of its complications (hyponatremia, hepatic hydrothorax), hepatorenal syndrome, spontaneous bacterial peritonitis, and hepatic encephalopathy. SUMMARY: Important studies, mostly prospective, regarding the management of the complications of portal hypertension are reviewed, including trials that demonstrate the value of the hepatic venous pressure gradient in predicting these complications, a trial of beta-blockers in patients with small varices, a randomized trial of transjugular intrahepatic portosystemic shunt using covered stents and another pilot study using this shunt in the treatment of hepatorenal syndrome, a trial of antibiotic prophylaxis in preventing early variceal rebleeding, and a trial of synbiotic therapy in hepatic encephalopathy. These trials will contribute to advancing the practice of hepatology and defining future research areas. PMID- 15818153 TI - Alcohol and the liver. AB - PURPOSE OF REVIEW: To highlight salient recent discoveries and results of clinical trials in alcoholic liver disease (ALD). The burden of care for ALD patients is hefty and the prevalence of alcohol abuse may be increasing in both the developed and the underdeveloped world. RECENT FINDINGS: Molecular mechanisms of alcoholism are being identified but not of the predisposition to alcoholic liver injury, except perhaps for polymorphism of a cytotoxic T-cell antigen. The Mayo End-stage Liver Disease (MELD) score performs well in assessing the prognosis of ALD; serological biomarkers for predicting ALD outcome are of uncertain value. Concomitant liver disease (e.g., obesity, hepatitis C, and iron overload) aggravates the severity of ALD; conversely, alcohol abuse may be a cryptic co-factor in some cases of non-alcoholic fatty liver. For alcoholic hepatitis, nutritional support is the mainstay of treatment; steroids are considered by some (but not all) as safe and effective therapy, whereas manipulations of tumor necrosis factor-alpha activity have been disappointing, or of unproven benefit at best. In liver transplantation for ALD, methods are being devised to monitor recidivism and to ameliorate its risk and that of co-morbid psychiatric conditions. SUMMARY: Much of the pathogenesis of ALD has been identified and headway has been made in predicting its prognosis. However, much remains to be done to elucidate the molecular genetics of the risk of developing ALD and in formulating safe, effective therapies for alcoholic hepatitis. PMID- 15818154 TI - Liver transplantation. AB - PURPOSE OF REVIEW: Liver transplantation continues to change as we further define appropriate criteria for allocation and utilization of this scarce resource. The following review highlights new trends and ideas in this evolving field. RECENT FINDINGS: Although the model for end-stage renal disease (MELD) scoring system appears to fairly accurately predict mortality while waiting for transplant, the system may be less accurate in predicting outcomes following transplantation. MELD scores offer an additional advantage to patients with hepatocellular carcinoma (HCC), bringing them to transplant sooner with overall better survivals. However, despite its advantages, the MELD scoring system does not resolve the disparity in the allocation of organs between various organ procurement organizations. Several variables appear to affect patients with hepatitis C undergoing liver transplantation. Selection of appropriate donors appears to be important when transplanting patients with hepatitis C virus (HCV) infection as increasing donor age is associated with poorer outcomes. However, the controversy over whether a living donor liver transplant (LDLT) results in poorer outcomes in HCV infected patients remains. Post-transplant medical treatment of HCV may result in both a sustained virologic response and improved histology. With improved overall survival in patients undergoing orthotopic liver transplant (OLT), increasing attention has been focused on the medical complications following transplant. Identifying specific contributing factors in the development of renal dysfunction and devising strategies to prevent its occurrence are critical to further improvements in outcome following OLT. SUMMARY: As the gap between patients and available organs remains, continued investigation into appropriate allocation and maximization of outcomes following liver transplant will continue. PMID- 15818155 TI - Cholangiocyte biology. AB - PURPOSE OF REVIEW: Cholangiocytes are increasingly recognized as biologically important because of the diverse array of cellular processes in which they participate. Collectively, these processes define normal function and, when disturbed, account for abnormalities that cause disease. Advances in animal models and sophisticated technology in imaging and gene silencing have allowed progress in defining the roles that cholangiocytes play in signaling, transport of water, ions and solutes, and alterations that result in cholestasis. The pace of advances in technology justifies a yearly summary to identify the most important developments in cholangiocyte biology. RECENT FINDINGS: The main areas of recent progress include insights into the molecular mechanisms of bile secretion and the development of new experimental models and technologies. SUMMARY: Understanding the critical components and key biologic processes in cholangiocytes responsible for regulation of ductal bile secretion is an initial and required step in generating hypotheses relevant to disease. With regard to the pathologic relevance of this work, cholestatic liver diseases represent a broad group of hepatobiliary disorders with which hepatologists must deal. In addition to genetic defects, the study of the normal and altered trafficking of cholangiocyte transport systems involved in bile secretion may provide a molecular correlate for the functional changes that occur in disease. Critical to this understanding is the ongoing development of experimental models and techniques to interpret data to answer key hypothesis-driven questions. Second, the collegial sharing and exchange of novel concepts, ideas, reagents, and probes promotes positive advances in the field. PMID- 15818156 TI - Pediatric hepatobiliary disease. AB - PURPOSE OF REVIEW: This review summarizes recent advancements occurring over the past year in the field of pediatric hepatology. They have helped in the understanding and treatment of several liver and biliary tract disorders of childhood. RECENT FINDINGS: Recent advancements in the areas of childhood primary sclerosing cholangitis, nonalcoholic fatty liver disease, acute liver failure, liver transplantation, neonatal hemochromatosis, and the Biliary Atresia Research Consortium have been summarized. SUMMARY: Developments and continued research in these areas have the potential to bring significant benefits to children affected by these disorders. PMID- 15818157 TI - Sclerosing cholangitis. AB - PURPOSE OF REVIEW: Primary sclerosing cholangitis is a chronic cholestatic liver disease characterized by strictures of the biliary tree. It is immune mediated, although the precise cause remains unknown. Recent reports have shown a higher prevalence and burden of disease than was previously suspected. RECENT FINDINGS: The research into the etiopathogenesis, epidemiology, diagnosis of cholangiocarcinoma, medical and surgical therapy, and timing and outcome of liver transplantation is discussed. SUMMARY: Genetic heterogeneity among patients with primary sclerosing cholangitis is supported, and further gene polymorphisms associated with protection against primary sclerosing cholangitis have been elucidated. Bile duct injury seems to be a multistep process. Magnetic resonance cholangiopancreatography is a cost-effective and accurate way of diagnosing primary sclerosing cholangitis in comparison with endoscopic retrograde cholangiopancreatography. Ursodeoxycholic acid may have a role as a colorectal and hepatobiliary cancer chemopreventive agent. Liver transplantation remains the only treatment in end-stage disease. The 5-year and 10-year patient and graft survival rates are comparable with those in patients without primary sclerosing cholangitis, but there is a higher rate of retransplantation for primary sclerosing cholangitis in most centers. Hepatobiliary malignancy is found in a minority of patients at transplantation, although 5-year survival rates for these patients are still promising. PMID- 15818158 TI - Update on biliary endoscopy. AB - PURPOSE OF REVIEW: Since the introduction of endoscopic retrograde cholangiopancreatography, there have been great improvements in the technique, equipment, and clinical utility of biliary endoscopy. In many cases, therapeutic endoscopic retrograde cholangiopancreatography can take the place of invasive surgery, including common bile duct exploration, thereby decreasing the patient's morbidity and recuperation time. Even with advances, such as stent placement for biliary strictures in a patient after liver transplantation, there is still room for improvement. RECENT FINDINGS: Research over the past year has been focused on areas including improved imaging and tissue sampling of the biliary tree through endoscopic ultrasound techniques, better identification of patients at risk for pancreatitis after endoscopic retrograde cholangiopancreatography, and refinements in photodynamic therapy for the treatment of cholangiocarcinoma. International research has continued to emphasize biliary sphincterotomy compared with balloon dilation for the management of choledocholithiasis. Further developments in other fields, such as living related liver transplantation, provide challenges for the biliary endoscopist. SUMMARY: This review focuses on some of the work being performed in the field of biliary endoscopy. The key studies were chosen to highlight some of the areas currently being investigated as well as to indicate certain fields that need further development. PMID- 15818159 TI - The role of toll-like receptor 9 in the intestine. PMID- 15818160 TI - Bibliography. Current world literature. PMID- 15818162 TI - Molecular markers, molecular-targeted therapies and taxanes: how to integrate the progress into clinical research and practice for the management of head and neck cancers. PMID- 15818163 TI - Monoclonal antibodies targeting epidermal growth factor receptor and vascular endothelial growth factor with a focus on head and neck tumors. AB - PURPOSE OF REVIEW: To provide an overview of recent clinical trials with monoclonal antibodies targeting epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) in head and neck cancer (HNC) and in other tumors. To discuss future therapeutic strategies. RECENT FINDINGS: Cetuximab, a chimeric monoclonal antibody targeting EGFR, has induced improved locoregional control and survival in combination with radiotherapy in a phase III study in locally advanced inoperable HNC. The recent Bowel Oncology with Cetuximab Antibody (BOND) study has shown that the combination of irinotecan and cetuximab yields a better response rate and a longer time to progression with respect to cetuximab monotherapy in irinotecan-refractory metastatic colorectal cancer, pointing to both a cetuximab single-agent activity and a cetuximab potential for reversal of irinotecan resistance. Non-small cell lung cancer and pancreatic cancer represent further areas for cetuximab development. Bevacizumab is a humanized monoclonal antibody that targets VEGF. It is the first antiangiogenic drug to have induced a survival advantage in cancer therapy, within a randomized trial of irinotecan, 5-fluorouracil, leucovorin (IFL) combined with bevacizumab or placebo in metastatic colorectal cancer. The use of bevacizumab in HNC is supported by preclinical evidence of an angiogenic loop; a few clinical trials are exploring the feasibility and the therapeutic potential of a combination of bevacizumab and EGFR-targeted drugs. SUMMARY: Monoclonal antibodies targeting EGFR and VEGF represent exciting therapeutic strategies that should be further evaluated both in combination with drugs acting on the same target at a different level and in combination with other antisignaling agents acting on different pathways. PMID- 15818164 TI - Taxanes in the treatment of head and neck cancer. AB - PURPOSE OF REVIEW: This review presents new data on the role of paclitaxel and docetaxel in the management of squamous cell carcinoma of the head and neck. Recently both agents have been tested in squamous cell carcinoma of the head and neck in combination with other chemotherapeutic agents, targeted drugs, and radiotherapy in in-vitro experiments and in the clinic as first-line treatment of patients with metastatic/recurrent and locally advanced squamous cell carcinoma of the head and neck. RECENT FINDINGS: The combination of taxanes with standard or accelerated radiotherapy is feasible and induction chemotherapy followed by chemoradiation is active and feasible without excessive toxicity in patients with locally advanced squamous cell carcinoma of the head and neck. The use of low dose fractionated radiotherapy shows promising in-vitro and clinical results and is further explored. SUMMARY: Both docetaxel and paclitaxel can be combined with chemotherapeutic agents and radiotherapy, but phase III studies are needed to prove the superiority of these approaches compared to standard treatment. The final results of the combination study of cisplatin and 5-fluorouracil with or without docetaxel may change the standard chemotherapeutic regimen for induction chemotherapy in patients with locally advanced squamous cell carcinoma of the head and neck. PMID- 15818165 TI - Advances in nasopharyngeal carcinoma. AB - PURPOSE OF REVIEW: Intergroup 0099 established the role of concurrent chemotherapy with radiation therapy in the treatment of locally advanced nasopharyngeal carcinoma, but its reproducibility was unclear and chemotherapy compliance was poor. Multiple concurrent chemoradiation phase III trials were initiated in response to the Intergroup trial, and technologic advances in radiotherapy were explored to improve the therapeutic ratio. This review highlights recent advances in the management of nasopharyngeal carcinoma as a result of these endeavors. RECENT FINDINGS: Five randomized phase III trials confirmed the benefit of concurrent chemoradiation over radiation therapy alone, firmly establishing concurrent chemoradiation as the standard of care in locally advanced nasopharyngeal carcinoma. Each of these studies used conventional radiation therapy and noted an increase in toxicity over radiation therapy alone. Intensity-modulated radiation therapy is an advanced form of three-dimensional conformal radiotherapy which allows delivery of high doses of radiation to the tumor while sparing adjacent normal tissues, leading to improved local control and decreased radiation therapy-induced toxicities. Distant metastasis remains a significant problem despite intensity-modulated radiation therapy. Taxane-based induction chemotherapy seems promising in phase II studies. Targeted therapies remain a major area of interest and require further investigation. SUMMARY: Cisplatin-based concurrent chemoradiation followed by adjuvant chemotherapy is the standard of care for locally advanced nasopharyngeal carcinoma. Intensity modulated radiation therapy has undergone a rapid evolution and is replacing conventional radiation therapy in many institutions. A multidisciplinary effort is under way to explore more effective systemic therapy to improve the distant metastasis free rates. PMID- 15818166 TI - Intensity-modulated radiation therapy in the management of head and neck cancer. AB - PURPOSE OF REVIEW: Intensity-modulated radiation therapy (IMRT) represents a potentially significant new advance in the radiotherapeutic treatment of head and neck cancer patients. The capacity of IMRT to produce highly conformal dose distributions affords the opportunity to decrease the spectrum of toxicities associated with head and neck radiation. This review updates recent advances regarding the use of this emerging technology. RECENT FINDINGS: Mature clinical data demonstrating head and neck IMRT safety and efficacy remain relatively limited to date. Recent publications have sought to clarify target selection and delineation. Refinements in target design driven by clinical outcomes are beginning to emerge. Technical aspects of IMRT delivery such as the impact of daily set-up variation, planning methods, and optimal fractionation regimens are progressing. SUMMARY: Stepwise refinement in the practice of IMRT for head and neck cancer patients is advancing worldwide. However, mature clinical data remains relatively limited and specific aspects regarding the practice of head and neck IMRT remain heterogeneous. PMID- 15818167 TI - Low-level laser for prevention and therapy of oral mucositis induced by chemotherapy or radiotherapy. AB - PURPOSE OF REVIEW: Oral mucositis is a common morbid condition associated with chemotherapy or radiotherapy for which there is no standard prophylaxis or treatment. There is increasing evidence that the use of low-level laser can reduced the severity of mucositis associated with chemotherapy or radiation therapy. The purpose of this review is to examine the available evidence for it. RECENT FINDINGS: For most approaches commonly used to prevent or treat chemotherapy-associated or radiotherapy-associated oral mucositis, a recent panel of experts could not find sufficient levels of evidence to recommend or suggest their use. As for low-level laser therapy, the results are difficult to assess and compare because of interoperator variability and because clinical trials are difficult to conduct in that field. Nevertheless, there is accumulating evidence in support of low-level laser therapy. SUMMARY: On the basis of literature data, it is reasonable to conclude that the evidence that low-level laser therapy may be useful in decreasing the severity of chemotherapy-associated or radiotherapy associated mucositis is substantial, even though there have been few controlled studies in the field of prevention. PMID- 15818168 TI - The evolving role of surgery in the management of head and neck tumors. AB - PURPOSE OF REVIEW: This paper reviews recent surgical contributions to the multidisciplinary approach to head and neck cancer treatment as well as published data on pertinent questions in the field. RECENT FINDINGS: The role of surgery in the strategy of organ preservation via chemoradiation, including treatments of complications and locoregional recurrences, such as when N1-N3 necks should be operated on and the role of CT and positron emission tomography (PET) scanning. Also covered are the role of sentinel lymph node biopsy for the detection of occult micrometastases in the N0 neck and the value of fluorodeoxyglucose PET scanning combined with the former. Transoral laser surgery for tumors of the larynx or of the hypopharynx is discussed, as is a new technique of reconstruction involving the anterolateral thigh free flap. Also included are quality of swallowing after resections and reconstruction of the oral cavity and orpharyngeal tumors. SUMMARY: Surgery must define its role in the multidisciplinary treatment of advanced cancers, which currently often favors (chemo)radiotherapy protocols. Organ sparing by transoral laser surgery for laryngeal and hypopharyngeal cancer has established itself within our armamentarium and its exact role will be further refined. Larger and well executed studies on sentinel lymph node biopsy in the clinically negative neck are needed before this procedure can be implemented in our routines. The latest addition, the anterolateral thigh flap, to further decrease patients' morbidity is unlikely to completely replace the currently favored radial forearm flap in soft tissue replacement of the head and neck. Outcome analysis, including swallowing, is a major step that surgeons should further investigate. PMID- 15818169 TI - 18F-fluoro-2-deoxy-D-glucose positron emission tomographic imaging: recent developments in head and neck cancer. AB - PURPOSE OF REVIEW: Positron emission tomography using 18F-fluoro-2-deoxy-D glucose (18FDG-PET) is well established in clinical routine as a metabolism-based whole-body imaging tool for cancer diagnosis and follow-up. Several reports have appeared indicating the potential and limitations of this technique in head and neck cancer (HNC). This review limits its scope to the recent advances using 18FDG-PET in the clinical management of HNC. RECENT FINDINGS: The combination of 18FDG-PET and sentinel node biopsy has been explored for the surgical treatment planning of oral and oropharyngeal cancer. Recent reports indicate that multimodality imaging combining PET with high-end CT scanning increases the diagnostic accuracy. 18FDG-PET has a potential for use in radiation treatment planning and for the prediction of response and early evaluation of treatment efficacy. SUMMARY: Increasingly 18FDG-PET is used as a clinical imaging modality in the different stages of the management of HNC. In particular, its clinical value in initial staging of neck lymph nodes and in the evaluation of recurrent or residual disease is well established. In these settings 18FDG-PET has been shown to be more accurate than conventional imaging. Recent studies indicate that 18FDG-PET could be of additional value in staging the N0 neck, in radiation treatment planning, and in prediction of treatment efficacy. PMID- 15818170 TI - Questions without answers. PMID- 15818171 TI - Progress in detection and treatment of prostate cancer. AB - PURPOSE OF REVIEW: The purpose of this review is to highlight the most important developments in the diagnosis, prevention, and management of prostate cancer reported in the past year that have been published in the medical literature. RECENT FINDINGS: Recent research has shown that clinically meaningful prostate cancer can develop in patients with a PSA value <4 ng/dL and has suggested that 'watchful waiting' approaches may not be appropriate for young patients or those with a life expectancy of greater than 15 years. The contribution of obesity to high-grade disease and a higher rate of relapse has also been described. There is now randomized data suggesting that adjuvant hormonal therapy for as little as six month's time confers a survival benefit for those receiving radiation therapy. Secondary hormonal manipulation with ketoconazole is now supported by randomized phase III data. Most importantly, this year the results of two randomized phase III clinical trials of chemotherapy in hormone refractory prostate cancer have shown that survival can be increased by 20% with the use of docetaxel. SUMMARY: Recent data reported in the past year have significant implications for those involved in the management of prostate cancer. The results of the chemotherapy trials have resulted in new standards of care for this disease. PMID- 15818172 TI - Renal cell carcinoma. AB - PURPOSE OF REVIEW: This review focuses on recent developments in the biology and clinical therapeutics of renal cell carcinoma. Given historically limited advances in this disease, a more thorough understanding and testing of rationally targeted agents is needed. RECENT FINDINGS: Von Hippel-Lindau gene inactivation is observed in most clear cell renal carcinoma, driving the malignant phenotype. The resulting vascular endothelial growth factor overexpression has been targeted though various approaches, with a clear signal of anti-tumor activity. In addition, immunotherapy remains a therapeutic standard in renal cell carcinoma and an area of ongoing investigation. Observation of small renal masses may represent a viable clinical option. SUMMARY: Renal cell carcinoma has become a model disease for rationally targeted therapeutics based on significant understanding of the underlying biology. Recent advances have increased the potential for meaningful improvements in clinical outcomes for renal cell carcinoma patients. PMID- 15818173 TI - Germ cell tumors. AB - PURPOSE OF REVIEW: This review provides an overview of some of the recent pre clinical and clinical developments in germ cell tumors. RECENT FINDINGS: Recent epidemiological studies highlight the variations in the geographic and ethnic distribution of germ cell tumors and the changing incidence of seminoma versus nonseminoma in the population. Additional studies are continuing to identify risk factors for germ cell tumors. Expression profiling, both at the gene and protein levels, is beginning to identify, at the molecular level, some of the factors associated not only with germ cell pluripotency but also with the different histologic subtypes of germ cell tumors. Work in the area of identifying potentially new serum tumor markers in germ cell tumor, as well as the role of the traditional tumor markers in predicting outcome to therapy is ongoing. Data is emerging on the role of positron emission tomography in evaluating residual lesions in seminoma. Evolving data on chemotherapy, radiation, and surgery further complements and clarifies information on these treatment modalities, and their potential toxicities, in the management of germ cell tumors. SUMMARY: Ongoing preclinical and epidemiological studies highlight the complexities underlying germ cell tumor pathogenesis. With enhanced understanding of some of these processes, treatments, particularly for advanced stages, will continue to evolve. PMID- 15818174 TI - Bladder cancer. AB - PURPOSE OF REVIEW: This article reviews the recent literature concerning important issues in the management of patients with bladder cancer. A brief overview of all aspects of bladder cancer including the etiology, diagnosis, and treatment are discussed with a focus on recent advances. RECENT FINDINGS: Bladder cancer is a significant cause of morbidity and mortality. The treatment for bladder cancer should be based on individual patient risk assessment and should include a multidisciplinary approach. In patients with superficial bladder cancer, research has focused on improving and optimizing intravesical therapy to reduce tumor recurrence and progression as well as on methods to better select the most appropriate treatment for patients with high-risk features. The important prognostic and therapeutic role of lymphadenectomy during radical cystectomy has become apparent and recent work has attempted to better define what should be considered the standard for lymph node dissection. Finally, in an attempt to improve survival, advances have been made using systemic chemotherapy in both the perioperative settings as well as for treatment of metastatic bladder cancer. SUMMARY: Research continues to improve our understanding of bladder cancer. This ongoing investigation is currently being translated to the bedside with refinements in the diagnosis and treatment of patients with bladder cancer. PMID- 15818175 TI - Bibliography. Current world literature. Head and neck. PMID- 15818177 TI - Towards improving outcomes of fatal pulmonary infections. PMID- 15818178 TI - Recent advances in the diagnosis and management of tuberculosis. AB - PURPOSE OF REVIEW: Tuberculosis (TB) remains one of the leading infectious killers of adults in the world today. This paper will review recent advances and understanding in the epidemiology, diagnosis, and management of TB. RECENT FINDINGS: Tuberculosis remains a significant global threat, particularly in regions of the world heavily affected by HIV. Diagnosis of TB infection and disease rely on outdated and problematic methods, but newer immunologic and nucleic acid-based techniques are emerging. Treatment of latent TB infection still relies mainly on the use of isoniazid but several treatment-shortening strategies are being studied. Treatment of active disease has advanced little since the introduction of short-course chemotherapy with rifampin, but several new drugs are being developed and studied. Timely initiation of HIV treatment in co-infected patients is increasingly seen as important and strategies for initiating therapy for both diseases are being refined. The existence of immune reconstitution inflammatory reactions is also becoming more widely appreciated. Other populations at risk for TB such as those receiving TNF-alpha antagonists are being recognized and improved screening and control measures implemented. SUMMARY: The global epidemiology of TB has been shaped in recent decades by HIV, urbanization and poverty. Diagnosis and treatment remain challenging, however, improved screening of at-risk populations and new diagnostic modalities and treatment strategies offer hope to the millions who suffer from tuberculosis. PMID- 15818179 TI - Utility of the antigen-specific interferon-gamma assay for the management of tuberculosis. AB - PURPOSE OF REVIEW: The tuberculin skin test, now over a century old, is prone to reader variability, and outcomes are influenced by cross-reactivity with environmental mycobacteria, previous bacillus Calmette-Guerin (BCG) vaccination, and anergy in immunosuppressed individuals. More recently, T-cell-based interferon-gamma responses to Mycobacterium tuberculosis - specific antigens have been investigated for their role in diagnosing latent tuberculosis infection. RECENT FINDINGS: We review the evidence supporting the utility of the interferon gamma assay for the diagnosis of latent tuberculosis infection (LTBI) in low prevalence countries. We discuss the principle of the test, technical factors related to performance, and its utility in active tuberculosis, in specialised subgroups such as immunocompromised patients, and its applicability in developing countries. SUMMARY: Compared with the tuberculin skin test, the antigen-specific interferon-gamma assay, when used in a standardised protocol (overnight incubation assay using a combination of two antigens) for the diagnosis of LTBI, has greater specificity in BCG-vaccinated individuals, displays a stronger association with exposure, and is less biased by environmental mycobacteria such as Mycobacterium avium. Prospective studies are required, however, to confirm that treating LTBI, as defined by the interferon-gamma assay, will reduce the tuberculosis burden in low-prevalence countries and whether interferon-gamma responses are predictive of those who have a high risk of progression to active tuberculosis. Further studies are required to address the utility of the interferon-gamma assay in specialised subgroups of patients, in developing countries, and as a marker of disease activity. PMID- 15818180 TI - Infectious lung complications in patients with HIV/AIDS. AB - PURPOSE OF REVIEW: The aim of this article is to review recent observations in the area of infectious lung complications in individuals with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS). Since the immunodeficiency was first characterized, it has been associated with enhanced susceptibility to opportunistic infection, and life-threatening infections of the lung in particular. In the past few years there have been a large number of reports documenting the changes to this disease profile in the age of highly active antiretroviral therapy (HAART). Furthermore, there have been considerable advances in our understanding of the immunology and vaccinology of many of the pathogens implicated in pulmonary infections in this context, including Mycobacterium tuberculosis, Streptococcus pneumoniae and Pneumocystis pneumonia. RECENT FINDINGS: In considering the time-course and spectrum of HIV-associated respiratory infections, the field must now be divided into studies undertaken in parts of the world where HAART is accessible and those where it is not. Despite the enormous impact of HAART, it has brought with it a new set of concerns, including the effects of immune restoration disease (IRD), and the complex interplay between HAART and tuberculosis therapy. SUMMARY: The overriding conclusion from recent experience is that HAART, in those parts of the world where it is readily available, has changed the clinical picture of infections associated with HIV, and needs to be available to patients across the developing world as well. Furthermore there have been important developments in vaccination programs against pathogens involved in HIV-associated pneumonia. PMID- 15818181 TI - Noninfectious pulmonary complications of HIV/AIDS. AB - PURPOSE OF REVIEW: This article reviews recent findings on noninfectious pulmonary complications of HIV/AIDS, with a focus on HIV/AIDS-related lung malignancies and pulmonary hypertension, and discusses their incidence in the highly active antiretroviral therapy (HAART) era. RECENT FINDINGS: Noninfectious pulmonary complications of HIV/AIDS are now recognized as important contributors to morbidity and mortality in HIV-infected patients. This is especially the case for HIV-related lung cancer and other non-AIDS-defining malignancies, which are now being diagnosed with increased frequency in HIV-infected patients. The incidence of Kaposi sarcoma and AIDS-related lymphoma has decreased in the HAART era, but compared with the general population, the risk of these malignancies and pulmonary hypertension is still very high in HIV-infected patients. Concurrent use of HAART and chemotherapy improves prognosis and survival of patients with AIDS-related lymphoma. For patients with HIV-related pulmonary hypertension, some studies show no beneficial effect of HAART whereas other reports show that HAART improves patient survival and response to antihypertensive treatment. SUMMARY: The beneficial effect of HAART and improved immune response on the treatment of Kaposi sarcoma and AIDS-related lymphoma suggests that HIV or viral-induced immunosuppression plays an important role in the development of these malignancies. Evidence from current studies suggests that HAART does not protect against HIV-related lung cancer. The full impact of HAART on HIV pulmonary hypertension remains to be determined. PMID- 15818182 TI - Pulmonary infections in non-HIV-immunocompromised patients. AB - PURPOSE OF REVIEW: Pulmonary infections are the most frequent complications in non-HIV-immunocompromised patients and portend a high mortality. This scenario represents a challenging task for clinicians and an important subject of clinical research from different perspectives. This review comments on the results of relevant original articles in this area published from 2003 to the present. RECENT FINDINGS: The present review addresses the etiology of the pulmonary infiltrates in immunocompromised patients, the use of new emerging diagnostic tools and medical devices in the clinical management of these infiltrates, and the greater understanding of the inflammatory immune response associated with infection in this setting. SUMMARY: Advances in diagnostic tests and therapeutic devices are facilitating the clinical management of pulmonary infections. New challenges are emerging, however, such as the growing evidence regarding the important role of respiratory viruses as a common cause of lower respiratory tract infections. Finally, new insights into the mechanisms of the inflammatory response associated with pulmonary complications can help understanding their pathogenesis, improve prevention and diagnosis, and anticipate future therapeutic modalities. PMID- 15818183 TI - Community-acquired pneumonia in children. AB - PURPOSE OF REVIEW: This review highlights recent developments in the diagnosis, etiology, therapy, and prevention of community-acquired pneumonia in children. RECENT FINDINGS: Sensitive new diagnostic methods have increased the detection rate of the causative agent up to 94%. Streptococcus pneumoniae is the most prevalent bacterial pathogen in all ages. Polymerase chain reaction is a rapid and sensitive method for the detection of Chlamydia pneumoniae and Mycoplasma pneumoniae, which have gained greater importance in recent years. During the period covered by this review, two new agents causing pneumonia were extensively studied. Human metapneumonovirus detected in young children is a leading cause of respiratory disease during the first years of life. A novel coronavirus was identified as the causative agent of severe respiratory syndrome, a new respiratory illness that affects adults and children. One multicenter trial concluded that nonsevere pneumonia can be treated with a short course of oral amoxicillin and a multicenter international study showed that children with severe pneumonia have similar outcomes whether treated with oral amoxicillin or parenteral penicillin, but more data are needed to demonstrate the safety and efficacy of such regimens. SUMMARY: The continued evolution of bacterial resistance highlights the need for appropriate use of antibacterials. Improved diagnostic techniques will aid the treatment of children with community-acquired pneumonia. Aggressive vaccination with the pneumococcal conjugate vaccine and other available vaccines as well as the development of new vaccines will aid the prevention of respiratory disease in children. PMID- 15818184 TI - Pneumonia in the elderly (geriatric) population. AB - PURPOSE OF REVIEW: Review of recent studies (2003 and 2004) concerning pneumonia in the very old. RECENT FINDINGS: Hospitalisation for community-acquired pneumonia (CAP) in the elderly is associated with a high mortality and with a high rate of readmission within the following year. Functional status, altered mental status, number of comorbidities, aspiration pneumonia, renal failure, and nutritional status are all indicators of adverse prognosis. Although Streptococcus pneumoniae, Haemophilus influenzae, Enterobacteriacae, and Staphylococcus aureus are the most frequently identified causative microorganisms, viruses account for up to 26% of hospital admissions for CAP. Chlamydia pneumoniae is also implicated in CAP and in nursing-home-acquired pneumonia (NHAP), with recent reports of outbreaks in nursing homes. Aspiration pneumonia is frequent in the elderly and occurs with increased frequency in patients with nasogastric tubes or percutaneous enterogastric tubes. In severe aspiration pneumonia, 20% of organisms implicated are anaerobic and 80% aerobic, most of which are gram-negative Enterobacteriaceae. Poor oral hygiene increases subsequent risk of pneumonia: dental plaque may act as a reservoir for pathogenic organisms implicated in CAP or NHAP. Prevention of CAP and NHAP relies on the combined use of influenza and pneumococcal vaccination, which decreases hospital admissions and in-hospital mortality for CAP. SUMMARY: Recent studies stress the importance of aspiration as a frequent mechanism of CAP, provide new insights as to causative organisms in this setting, and underline the contribution of combined vaccination in reducing morbidity and mortality. PMID- 15818185 TI - Nosocomial pulmonary infections in HIV-positive patients. AB - PURPOSE OF REVIEW: Nosocomial infections (NI) constitute a significant public health problem and contribute to prolonged hospitalization, additional healthcare costs, and excess morbidity and mortality. Immunocompromised patients, including HIV-infected individuals, are at increased risk for NI, and 15-18.3% of them are represented by lower respiratory tract infections. Nosocomial pulmonary infections (NPI) appear to be more common in patients with acquired immunodeficiency syndrome (AIDS), as a result of the degree of immunosuppression, prior use of antibiotics, and exposure to invasive procedures. RECENT FINDINGS: This article reviews the epidemiologic and clinical evidences and reports on the occurrence of NPI in HIV-infected inpatients. SUMMARY: Although underestimated, NI occur commonly in HIV-infected patients, and among them nosocomial pneumonia, including tuberculosis and bacterial pneumonia, are associated with significant morbidity and mortality. The improvement of antiretroviral therapeutic options in developed countries has resulted in a decreased hospitalization rate of HIV infected individuals. Healthcare delivery in the in- and outpatient setting represents a potential for infections, including lower respiratory tract ones, according to the degree of immunosuppression and the intensity of invasive procedures. To minimize the risk of acquisition of healthcare associated low respiratory tract infections, adherence of healthcare workers to common infection practices, specific respiratory precautions, and early identification of persons who have tuberculosis or are at high risk for active tuberculosis, should be strengthened. PMID- 15818186 TI - Ventilator-associated pneumonia. AB - PURPOSE OF REVIEW: This review summarises some of the notable papers on ventilator-associated pneumonia (VAP) from January 2003 to October 2004. RECENT FINDINGS: Ventilator-associated pneumonia remains an important drain on hospital resources. All population groups are affected, but patients with VAP are more likely to be older, sicker, and male, with invasive medical devices in situ. Early VAP diagnosis is desirable to reduce VAP mortality and to retard emergence of multidrug-resistant microbes. This may be possible using preliminary culture results or intracellular organism in polymorphonuclear cells. In most intensive care units, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii are the commonest organisms isolated in VAP. However, causative organisms vary between and within hospitals. Consequently, individual intensive care units should develop empirical antibiotic policies to target the pathogenic bacteria prevalent in their patient populations. Preventative strategies aimed at reducing aerodigestive tract colonisation by pathogenic organisms, and also their subsequent aspiration, are becoming increasingly important. Educating medical staff about these simple measures is therefore pertinent. To reduce the occurrence of multidrug-resistant organisms, limiting the duration of antibiotic treatment to 8 days and antimicrobial rotation should be contemplated. Empirical therapy with antipseudomonal penicillins plus beta-lactamase inhibitors should be considered. If methicillin-resistant Staphylococcus aureus VAP is a possibility, linezolid may be better than vancomycin. SUMMARY: Prevention remains the key to reducing VAP prevalence. PMID- 15818187 TI - Pulmonary fungal infections. AB - PURPOSE OF REVIEW: Invasive fungal infections of the lung have historically been associated with an extremely high mortality. This review aims to disseminate the most recent advances in the diagnosis and management of fungal infections of the lung. RECENT FINDINGS: The number and diversity of immunosuppressed populations are growing rapidly. Transplant immunosuppression is becoming more aggressive early in the posttransplant period, potentially increasing the risk of invasive fungal infections. The galactomannan antigen test and the beta-D-glucan test have emerged as methods of serially monitoring at-risk patients for invasive aspergillosis. Their utility has been established in some neutropenic populations but not in solid organ transplant recipients. In-vitro studies, animal studies, and retrospective human studies support the use of combination antifungal therapy for invasive aspergillosis. Unfortunately no randomized clinical trials exist. SUMMARY: Invasive pulmonary aspergillosis will continue to be a major problem in immunocompromised patients in the future. Immense advances in the last 2-3 years are sure to improve outcome. Well-designed multicenter evaluations are still necessary, however, to optimize management as management options widen. PMID- 15818188 TI - Progressive and nonresolving pneumonia. AB - PURPOSE OF REVIEW: Patients with progressive and/or nonresolving community acquired pneumonia are at risk for increased morbidity and mortality. It is critical to be able to identify patients at risk to institute early appropriate therapy. The purpose of this review is to summarise the most updated developments in this area. RECENT FINDINGS: This review will glean from the recent literature clinical, laboratory, and radiologic findings that help identify patients at risk for such complications of their pneumonia. New studies will be reviewed that have identified some of the causes for treatment failures including the type of pathogen and discordant antimicrobial therapy. It will also discuss newly recognised and emerging infectious diseases that may result in progressive or nonresponding pneumonia including severe acute respiratory syndrome, avian influenzae, severe group A streptococcal disease, and community-acquired methicillin-resistant Staphylococcus aureus. Promising treatments have been identified for patients with progressive pneumonia due to an overwhelming host immune response including activated protein C and intravenous immunoglobulin. SUMMARY: Both progressive and nonresolving pneumonia represent treatment failure as a result of inappropriate initial therapy, a noninfectious cause, or an overwhelming immune response. It is critical to be able to identify patients with nonresponding pneumonia and to identify patients at risk for progressive pneumonia to institute appropriate therapy. PMID- 15818189 TI - Bibliography. Current world literature. Infectious diseases. PMID- 15818190 TI - Cardiac rehabilitation and exercise training programs in metabolic syndrome and diabetes. PMID- 15818191 TI - Commentary: frequently asked questions ... action packed answers. PMID- 15818192 TI - Prevalence of depressive disorders in men and women enrolled in cardiac rehabilitation. AB - PURPOSE: Epidemiological studies have demonstrated that depression is an independent risk factor for the development and recurrence of coronary heart disease (CHD). The prevalence of depressive disorders, however, is not well documented in CHD patients enrolled in cardiac rehabilitation (CR). The purpose of this study was to estimate the prevalence of current and lifetime depressive disorders in the CR setting. METHODS: One hundred ten men and women diagnosed with CHD and enrolled in a phase II CR program were screened via a psychiatric, structured interview to assess current and lifetime history of major depressive disorder, minor depression, and dysthymic disorder. RESULTS: A total of 17 (15.5%) individuals screened positive for a current depressive disorder at entry into the CR program, with 10 (9.1%) individuals screening positive for major depressive disorder, 7 (6.4%) for minor depression, and 6 (5.5%) for dysthymic disorder. With respect to lifetime prevalence of mood disorders, 29 (26.4%) individuals met the diagnostic criteria for at least one depressive disorder during their lifetime. The lifetime prevalence of major depressive disorder, minor depression, and dysthymic disorder was 11.8%, 14.5%, and 10.9%, respectively. Female CR participants evidence significantly higher current and lifetime prevalence rates for depressive disorders compared to their male counterparts. CONCLUSIONS: The results of this study suggest that a substantial number of CHD patients enrolled in CR report a clinically significant history of depression. Efforts to assess and treat depression are needed in the CR setting and may be associated with better adherence to lifestyle modification programs. PMID- 15818194 TI - Depression and cardiac risk. PMID- 15818195 TI - Geographic distribution of cardiac rehabilitation programs in the United States. PMID- 15818196 TI - Addressing the problem of cardiac rehabilitation program distribution. PMID- 15818197 TI - Nonparticipation in cardiac rehabilitation: an analysis utilizing classification and regression trees. PMID- 15818198 TI - Women's hearts--stress management for women with ischemic heart disease: explanatory analyses of a randomized controlled trial. AB - PURPOSE: This randomized controlled study aimed to evaluate the effects on psychosocial variables of a 1-year group-based cognitive-behavioral stress management program developed specifically for women with ischemic heart disease. METHODS: The present explanatory (per protocol) analyses include 80 women who were randomized to a 1-year cognitive-behavioral stress management program and 86 who were randomized to usual care (age = 35-77 years). Data were obtained before randomization and after 1 year, when the intervention group had completed the program. RESULTS: There were no statistically significant differences between the intervention and usual care groups in the psychosocial endpoints at randomization. Both groups improved in all psychosocial variables during the 1 year study period, but the rate of improvement was significantly greater in the intervention group for self-rated stress behavior (P = .006) and vital exhaustion (P = .03). Although changes were in favor of the treatment group also for depressive mood and quality of life, the rates of improvement between the 2 groups did not reach statistical significance (P = .23 and P = .10, respectively). CONCLUSION: A 1-year cognitive-behavioral stress management program designed specifically for women improved psychological well-being in some aspects in comparison with usual care. PMID- 15818199 TI - Long-term effects of cardiac rehabilitation and the paradigms of cardiac rehabilitation. AB - BACKGROUND: The benefits of exercise training for postmyocardial infarction and postcoronary artery bypass surgery patients are well established, but little is known about the effects of rehabilitation in the months or years following the program. The purpose of this study was to assess exercise capacity, blood lipids, and physical activity patterns 2 years after completing a concentrated residential rehabilitation program in Switzerland. METHODS: Seventy-eight patients (86% males, mean age = 56 +/- 10, mean ejection fraction = 64% +/- 12%) were referred to a residential rehabilitation program after a myocardial infarction or coronary artery bypass surgery between January 2001 and June 2001. Patients lived at the center for 1 month, during which time they underwent educational sessions, consumed a low-fat diet, and exercised 2 hours daily. Two years after completing the program, patients returned to the hospital and underwent a maximal exercise test, an assessment of recent and adulthood physical activity patterns, and evaluation of blood lipids. RESULTS: During the 2-year follow-up period, there were 5 deaths, and 70 of the remaining 73 patients returned for repeat testing. Mean exercise capacity increased 27% during the rehabilitation program (P < .01). Gains in exercise capacity during rehabilitation were maintained after the follow-up period; mean exercise capacity after 2 years was 34% higher compared with that at baseline (P < .01). At the 2 year evaluation, patients were expending a mean of 3127 +/- 1689 kcals/wk during recreational activities compared with 977 +/- 842 kcals/wk during adulthood prior to their cardiac event (P < .001). Between the completion of rehabilitation and the 2-year follow-up, total cholesterol, total cholesterol/high-density lipoprotein ratio, and triglycerides increased significantly. CONCLUSIONS: Two years after a cardiac event and participation in a concentrated residential rehabilitation program, patients maintained their exercise capacity and engaged in physical activities that exceed the levels recommended by guidelines for cardiovascular health. These observations suggest that a relatively intensive rehabilitation program provided a catalyst to maintain physical activity patterns and exercise tolerance in the 2 years following a cardiac event. PMID- 15818200 TI - Seattle Obstructive Lung Disease Questionnaire: sensitivity to outcomes in pulmonary rehabilitation in severe pulmonary illness. AB - PURPOSE: To evaluate the ability of the Seattle Obstructive Lung Disease Questionnaire (SOLDQ) to detect change in health-related quality of life (HRQOL) following a pulmonary rehabilitation program (PRP) and to determine associations with other functional indicators. METHODS: A prospective pre- and post-test design. A PRP was offered for 60 minutes, 2 times per week for 8 weeks. Measured at entry and exit to the PRP: the SOLDQ (physical, emotional, and coping skills), 6-minute walk, Medical Outcomes Study Short Form Health Survey-36 Veterans (SF 36V) (physical and mental), Lareau Functional Status and Dyspnea Questionnaire, and the Multidimensional Assessment of Fatigue (MAF) scale. Setting was an outpatient PRP in a VA Medical Center. RESULTS: Participants included 58 adults who completed the PRP (age mean = 66, SD +/- 9.8; FEV1% predicted mean = 39, SD +/- 15.9). At baseline, significant correlations were found between SOLDQ physical functioning and SF-36V physical, dyspnea, fatigue, 6-minute walk distance, SF-36V mental, and spirometry. SOLDQ emotional functioning significantly correlated with fatigue, dyspnea, and SF-36V mental and physical. SOLDQ coping skills significantly correlated with fatigue, dyspnea, and SF-36V mental and physical. Paired, 2-tailed, t tests comparing the SOLDQ subscales pre- and post-program were significant, with the biggest treatment effect seen with emotional functioning. Other measures showing significant improvement included 6 minute walk, SF-36V physical, dyspnea, and fatigue. CONCLUSIONS: After participation in a PRP, all 3 SOLDQ subscales improved significantly and were associated with other indicators of function. These findings validate the SOLDQ as a measure sensitive to functional changes associated with participation in pulmonary rehabilitation. PMID- 15818201 TI - Usefulness of the human activity profile, a functional performance measure, in people with chronic obstructive pulmonary disease. AB - PURPOSE: To further evaluate the usefulness of the Human Activity Profile (HAP) as a functional performance measure for those with chronic obstructive pulmonary disease (COPD). METHODS: Phase 1. The sample consisted of 53 outpatients (51 male) with stable COPD (forced expiratory volume in 1 second% predicted (FEV1% pred) = 38 +/- 14 (mean +/- standard deviation) at a university-affiliated veterans affairs medical center. Variables were functional performance (HAP Maximal Activity Score [MAS], HAP Adjusted Activity Score [AAS]), dyspnea (modified Borg category-ratio scale and Shortness of Breath Questionnaire [SOBQ]), and exercise capacity (6-minute walk distance [6MWD]). Phase 2. The HAP was administered pre-post pulmonary rehabilitation in 48 (14 male) outpatients with stable COPD (FEV1% pred = 35 +/- 15) at an outpatient pulmonary rehabilitation program at a large private hospital. RESULTS: Phase 1. Scores for MAS and AAS were, respectively, 62 +/- 12 and 44 +/- 15. The 6MWD, modified Borg, and SOBQ were, respectively, 1055 +/- 372 ft, 4.1 +/- 1, and 64.4 +/- 22 ft. Correlations of the 6MWD and dyspnea scores with the AAS were strong and explained 40% of the AAS variation. Phase 2. The baseline MAS and AAS scores were 55 +/- 14 and 42 +/- 15, respectively, with 29/48 (60%) classified as "low fitness" on the basis of the AAS. Post-rehabilitation, there was significant improvement in the AAS score (48 +/- 13) and significant reduction for those classified as "low fitness." CONCLUSIONS: The utility of the HAP as a measure of functional performance improvement in COPD is supported by its relationship with both 6MWD and dyspnea and its responsiveness to pulmonary rehabilitation. PMID- 15818203 TI - Unusual manifestations of metastatic pulmonary calcification: high-resolution CT and pathological findings. AB - The aim of this report is to describe the high-resolution CT and pathologic findings of 3 patients with unusual manifestations of metastatic pulmonary calcification. These include a case that presented with extensive dense consolidation, a case of metastatic calcification that improved spontaneously, and a case of metastatic calcification in a patient with no demonstrable biochemical abnormality or underlying disease. We conclude that metastatic calcification may present with dense bilateral consolidation, may improve spontaneously, and may rarely present in patients with no apparent underlying biochemical abnormality. PMID- 15818204 TI - Herpes simplex virus type 2 pneumonia after bone marrow transplantation: high resolution CT findings in 3 patients. AB - Bone marrow transplant recipients have increased prevalence of viral infections, including Herpes simplex pneumonia. The majority of infections are due to HSV type 1. We report the high-resolution CT findings in 3 bone marrow transplant patients with herpes simplex type 2 pneumonia. The most common CT features were focal areas of consolidation seen in 3 patients, and small centrilobular nodules and areas of ground-glass attenuation seen in 2 patients. PMID- 15818205 TI - Churg-Strauss syndrome: high resolution CT and pathologic findings. AB - OBJECTIVES: The purpose of this study was to evaluate high-resolution CT findings in 7 patients with Churg-Strauss syndrome and to compare the CT with the histopathologic findings. MATERIALS AND METHODS: High-resolution CT scans of 7 asthmatic patients (4 women, 3 men, age range, 34-62 years, mean 49 years) with Churg-Strauss syndrome were reviewed by 2 observers. Histologic specimens of lung obtained at surgical (n = 3) or transbronchial (n = 3) biopsy or autopsy (n = 1) were reviewed by an expert lung pathologist. The diagnosis of Churg-Strauss was based on clinical, laboratory, and histologic findings. RESULTS: Parenchymal and airway abnormalities included ground-glass opacities (n = 5), areas of air-space consolidation (n = 4), centrilobular nodules (n = 5), nodules 1-3 cm in diameter (n = 3), interlobular septal thickening (n = 4), bronchial wall thickening (n = 4), and areas of atelectasis (n = 1). Surgical biopsy (n = 3) and autopsy (n = 1) specimens demonstrated airspace disease in 3 patients, interlobular septal thickening in 3 patients, and airway abnormalities in 2 patients. Histologically, the airspace disease included eosinophilic pneumonia (n = 2) and small foci of organizing pneumonia (n = 1). The septal thickening was due to edema combined with numerous (n = 2) or few (n = 1) eosinophils. The airway abnormalities (n = 2) included muscle hypertrophy and large airway wall necrosis (n = 1) and eosinophilic infiltration of the airway walls (n = 1). Transbronchial biopsy (n = 3) demonstrated increased eosinophils. CONCLUSION: The main high-resolution CT findings of Churg-Strauss syndrome consist of airspace consolidation or ground glass opacities, septal lines, and bronchial wall thickening. These reflect the presence of eosinophilic infiltration of the airspaces, interstitium, and airways, and interstitial edema. PMID- 15818206 TI - Multislice CT evaluation of airway stents. AB - Tracheobronchial stents are playing an increasing role in the palliative treatment of large airway obstruction due to a variety of conditions, including extrinsic compression, intraluminal disease, and malacia. Computed tomography (CT) plays an important role in aiding planning of stent placement and in detecting various stent complications. In this pictorial essay, we illustrate and review the role of multislice CT in the pre- and post-stent placement settings. A special emphasis is placed upon the characteristic CT appearance of specific stent complications and upon the role of CT multiplanar reformations and 3 dimensional (3-d) reconstruction techniques. PMID- 15818207 TI - Spectrum of blunt chest injuries. PMID- 15818208 TI - Lymphangioleiomyomatosis: clinical, pathologic, and radiologic manifestations. PMID- 15818209 TI - Unusual late pulmonary complication in a child after umbilical cord blood transplantation: high-resolution CT-pathologic correlation. AB - We encountered a late pulmonary complication after umbilical cord blood transplantation (UCBT) that has not been previously reported. High-resolution CT (HRCT) findings of this disease were compared with the pathology. HRCT obtained on inspiration showed dilated thick-walled bronchioli, and innumerable centrilobular linear and branching structures in the bilateral middle and lower lobes. Neither mosaic perfusion nor air-trapping was seen in HRCT on inspiration and expiration. These HRCT findings were atypical compared with those of former bronchiolitis obliterans (BO) after bone marrow transplant (BMT). Pathologic specimens obtained by open lung biopsy showed thickening of the wall from the distal bronchioli to the alveolar ducts due to submucosal and intraepithelial infiltration of lymphocytes, histiocytes and foamy macrophages, which was not accompanied by organizing changes. These changes resemble lymphocytic bronchiolitis in lung transplant recipients, which was well correlated with HRCT findings. We think that our case was a new late pulmonary complication after UCBT. PMID- 15818210 TI - Cervical thoracic duct cyst. AB - Thoracic duct cysts are rare lesions presenting as mediastinal or supraclavicular masses. We report a case of a 77-year-old female who presented with a left supraclavicular mass. A cervical thoracic duct cyst was suspected after contrast enhanced computed tomography (CT) of the neck and chest. Diagnosis was confirmed by ultrasound-guided cyst aspiration and lymphangiography. Therapeutic embolization of the thoracic duct was unsuccessful. Definitive treatment was achieved with thoracic duct ligation using video-assisted thoracoscopic surgery (VATS) followed by surgical excision of the cyst. This is believed to be the first report of using minimally invasive surgery for thoracic duct ligation in the treatment of a thoracic duct cyst. PMID- 15818211 TI - Cystic bronchioloalveolar cell carcinoma. AB - Bronchioloalveolar cell carcinomas (BACs), a subset of primary lung adenocarcinomas, are uncommon. Similar to other non-small cell lung cancers, patients with BAC are usually 40-70 years of age. Distinguishing features relative to other non-small cell lung cancers include occurrence in young patients, a higher relative predominance in women, and weaker association with smoking. Histologically, they are a diverse group of malignancies that grow in a lepidic manner. Common radiologic presentations of BAC include a solitary nodule, airspace disease, and multiple nodules. Rare manifestations include cystic and cavitary disease. We report a case of a young woman with BAC that manifested as a chronic cough and bilateral air-containing opacities, initially misinterpreted as infection. Knowledge of the spectrum of radiologic manifestations of BAC and correlation with clinical history are important in suggesting the diagnosis and preventing misinterpretation. PMID- 15818212 TI - Large cryptococcoma mimicking lung cancer in an HIV-negative, type 2 diabetic patient. AB - Pulmonary cryptococcoma is an unusual disease in immunocompetent patients. The typical radiologic findings are single or multiple small nodules. We describe a case of a large pulmonary cryptococcoma in a 69-year-old patient, who presented with generalized weakness and poor appetite. Computed tomography of the chest showed a large mass that was strongly suggestive of lung cancer. Pulmonary cryptococcal infection was diagnosed by transthoracic needle biopsy. Human immunodeficiency virus testing was negative. The mass resolved with 200 mg daily of oral fluconazole over the course of 6 months. PMID- 15818213 TI - F-18 FDG uptake in a malignant localized fibrous tumor of the pleura. AB - In the case presented here, FDG-PET was performed to evaluate the possibility of malignancy. High FDG accumulation, with a standardized uptake ratio (SUR) of 3.0, was noted in an upper nodular compartment of the mass that exhibited malignant features histopathologically. It was suggested that FDG-PET is helpful to know which parts of lesions are benign or malignant in patients with LFTP whose prognoses are usually difficult to predict. PMID- 15818215 TI - Notes from the 2004 annual meeting of the Korean Society of Thoracic Radiology. PMID- 15818214 TI - Welder's lung associated with pneumothorax. AB - A 23-year-old male working in welding for 8 years was admitted to the hospital with chest pain. A large right pneumothorax with complete lung collapse was seen radiographically and a tube thoracostomy performed. At high-resolution computed tomography, multiple bilateral well-marginated nodules with surrounding ground glass opacity were seen. Welder's lung was considered in this patient because of his work in welding for 8 years with iron dust exposure, multiple nodules with ground-glass opacity were seen on HRCT, and serum ferritin and ferritin levels in bronchoalveolar lavage fluid were elevated. The granules in alveolar macrophages obtained from parenchymal biopsy and bronchoalveolar lavage stained strongly with iron dyes. Following avoiding exposure to welding fumes, the lung lesions disappeared without additional therapeutic interventions after a 3-week period. Welder's lung with associated lung lesions has been described, although complicating pneumothorax is unique to the case presented herein. PMID- 15818223 TI - What causes sudden hypokalemia? PMID- 15818226 TI - Pain management standards, the law, and you. PMID- 15818227 TI - Sounding the alarm for i.v. infiltration. PMID- 15818228 TI - Tailoring drug therapy with pharmacogenetics. PMID- 15818232 TI - Myths and factors...about endometriosis. PMID- 15818250 TI - Straight talk about ovarian cancer. AB - Learn how to help patients recognize and defeat this insidious malignancy. PMID- 15818252 TI - Mothering Becky. PMID- 15818253 TI - Patient education series. Lyme disease. PMID- 15818254 TI - Measure bladder volume without catheterization. AB - Learn how noninvasive ultrasound scanning can save your patient discomfort and infection risk. PMID- 15818255 TI - Assisted suicide: unraveling a complex issue. AB - If a terminally ill patient asked you for assistance in dying, how would you respond? Think about the ethical and legal considerations discussed here, then join the debate by sharing your opinions. PMID- 15818256 TI - Burned out? Here's help. PMID- 15818257 TI - When your patient has acute facial paralysis. AB - Learn to distinguish between Bell's palsy and Ramsay Hunt syndrome. PMID- 15818258 TI - 10 ways to help students grow. AB - Share your expertise with future nurses and watch them blossom-and prepare to reap the benefits of their enthusiasm. PMID- 15818259 TI - When your patient is in liver failure. AB - Find out what leads up to liver failure, how it's diagnosed, and how to care for your patient. PMID- 15818264 TI - When to use a midline catheter. PMID- 15818265 TI - Sizing up wounds accurately. PMID- 15818268 TI - Methamphetamine overdose. PMID- 15818271 TI - The importance of advanced directives. PMID- 15818272 TI - Evaluating chronic kidney disease risk. PMID- 15818274 TI - New rheumatoid arthritis treatments. PMID- 15818276 TI - Working out the complexities of severe sepsis. PMID- 15818277 TI - Asthma. PMID- 15818278 TI - NPs vs. IT for effective colposcopy patient education. PMID- 15818281 TI - Postauricular pain with Bell's palsy. PMID- 15818282 TI - Preventing teen hate crimes: our role as NPs. PMID- 15818287 TI - Randomized, investigator-blinded, multicenter study of gatifloxacin versus amoxicillin/clavulanate treatment of recurrent and nonresponsive otitis media in children. AB - BACKGROUND: Decreased susceptibility of pathogens to currently used agents for recurrent otitis media has provided the impetus for identifying new antimicrobial options. OBJECTIVE: To compare gatifloxacin with amoxicillin/clavulanate in children with recurrent or nonresponsive acute otitis media (AOM). METHODS: Included in this multicenter randomized trial were 413 patients, ranging in age from 6 months to 7 years, who had recurrent AOM (at least 3 episodes in the previous 6 months or 4 episodes in the previous 12 months) and/or had failed antibiotic therapy for AOM within 14 days of enrollment. Diagnosis required evidence of acute inflammation and otoscopic findings of middle ear effusion; baseline tympanocentesis was optional and encouraged. Children were randomly assigned (2:1) to 10 days of oral therapy with gatifloxacin suspension (10 mg/kg of body weight once daily) or amoxicillin/clavulanate suspension (45/6.4 mg/kg/d in 2 divided doses). RESULTS: : Clinical cure was obtained in 90.2% (222 of 246) of patients in the gatifloxacin group and 84.3% (102 of 121) of those in the amoxicillin/clavulanate group (95% confidence interval, -1.9-12.9) 3-10 days after treatment ended. Gatifloxacin was associated with higher clinical cure rates than was amoxicillin/clavulanate in children younger than 2 years of age (92.0% versus 80.0%, respectively). Cure rates by pretreatment pathogen in the gatifloxacin and amoxicillin/clavulanate groups were 92.1% (35 of 38) versus 88.9% (16 of 18) for Streptococcus pneumoniae infections and 88.2% (30 of 34) versus 92.3% (12 of 13) for Haemophilus influenzae infections, respectively. Sustained clinical cures 3-4 weeks after treatment ended were obtained in 74.4% (183 of 246) of patients treated with gatifloxacin and 72.7% (88 of 121) of those treated with amoxicillin/clavulanate. Adverse events considered drug-related occurred with similar frequency in the 2 groups. Six patients (2.2%) in the gatifloxacin group and 2 patients (1.5%) in the amoxicillin/clavulanate group developed transient symptoms of mild or moderate arthralgia. CONCLUSIONS: In this comparative evaluation of fluoroquinolone therapy in children with AOM, gatifloxacin was similar in clinical efficacy to amoxicillin/clavulanate 45/6.4 mg/kg/d for treatment of recurrent/nonresponsive infections. PMID- 15818288 TI - Randomized, investigator-blinded, multicenter, comparative study of gatifloxacin versus amoxicillin/clavulanate in recurrent otitis media and acute otitis media treatment failure in children. AB - BACKGROUND: Recurrent otitis media and acute otitis media treatment failure are commonly encountered in the pediatric population. OBJECTIVE: To compare the clinical efficacy of gatifloxacin with amoxicillin/clavulanate for the treatment of acute otitis media treatment failure and recurrent otitis media. METHODS: Three hundred fifty-four children 6 months-7 years with recurrent otitis media and/or acute otitis media failure were stratified according to age (younger than 2 years versus 2 years or older) and then randomly assigned to 10 days of treatment with gatifloxacin 10 mg/kg once daily or amoxicillin/clavulanate 90 mg/6.4 mg in 2 divided doses. Tympanocentesis was performed in 116 children with acute otitis media treatment failure and 52 with recurrent otitis media at study entry to validate the clinical diagnosis and provide microbiologic data. The primary outcome measure was clinical resolution of infection at the test-of-cure visit 3-10 days after completing treatment. RESULTS: Clinical resolution of acute otitis media was observed in 79.0% (49 of 62) of clinically evaluable children younger than 2 years and 90.3% (56 of 62) of those 2 years or older who were treated with gatifloxacin as compared with 77.6% (45 of 58) of children younger than 2 years and 79.7% (47 of 59) of children 2 years or older treated with amoxicillin/clavulanate. In patients with acute otitis media treatment failure, clinical response rates for children younger than 2 years and those 2 years or older were 87.5% (21 of 24) and 97.0% (32 of 33) with gatifloxacin versus 63.6% (14 of 22) and 83.9% (26 of 31) with amoxicillin/clavulanate. The corresponding clinical response rates in patients with recurrent otitis media were 79.2% (19 of 24) and 85.7% (18 of 21) with gatifloxacin and 90.5% (19 of 21) and 76.0% (19 of 25) with amoxicillin/clavulanate. Clinical success in those subjects having pretreatment middle ear fluid pathogens was similar for the 2 regimens [80.0% (24 of 30) gatifloxacin, 77.1% (27 of 35) amoxicillin/clavulanate]. Emergence of fluoroquinolone-resistant strains was not observed. Both drugs were generally well-tolerated. Diarrhea was the most common drug-related adverse event (10% gatifloxacin, 18% amoxicillin/clavulanate). No evidence of abnormal joint or gait findings was found during a 12-month follow-up. CONCLUSIONS: Gatifloxacin once daily is at least as effective and well-tolerated as amoxicillin/clavulanate twice daily in children with acute otitis media treatment failure or recurrent otitis media. There was no evidence of arthrotoxicity or emergence of fluoroquinolone-resistant bacteria in gatifloxacin-treated children. PMID- 15818289 TI - Risk factors for mortality resulting from bloodstream infections in a pediatric intensive care unit. AB - BACKGROUND: Bloodstream infections (BSIs) are prevalent in the critical care setting and have high attributable morbidity and mortality. The purpose of this study was to identify factors that significantly contribute to immediate as well as eventual mortality in patients with bloodstream infections at a pediatric intensive care unit (PICU). METHODS: Retrospective review of 2097 clinical records from admissions to our PICU in a 2-year period. Two separate case-control models were used. In the first model, eventual mortality (EM CASES) reflected those patients with eventual mortality, and EM CONTROLS were those who survived. In the second, infection-related mortality (IRM) cases were those with infection related mortality, defined as death within 7 days of BSI, and IRM CONTROLS were survivors past 7 days. Logistic regression was used to adjust for differences for 3 categories: patient characteristics, microbiology and treatment variables. RESULTS: We identified 74 separate episodes of bacteremia. Having an underlying malignancy or immunodeficiency was the only independently significant predictor of eventual mortality for BSI isolated within the PICU. Patients with infection related mortality more likely had Gram-negative bacteremia and/or fungemia, were older and had inadequate initial empiric antibiotic treatment at the time BSI was diagnosed. CONCLUSIONS: Targeted and aggressive early interventions should guide the empiric treatment of BSIs, whereas prolonged broad spectrum treatment should be minimized to avoid the emergence of resistant pathogen organisms. PMID- 15818290 TI - Postoperative mediastinitis in children: epidemiology, microbiology and risk factors for Gram-negative pathogens. AB - BACKGROUND: Mediastinitis, although an infrequent complication of median sternotomy, represents a significant source of morbidity and mortality. OBJECTIVE: To determine the incidence and describe the epidemiology and microbiology of mediastinitis in children after cardiac surgery and to identify risk factors for the development of Gram-negative mediastinitis. STUDY DESIGN: This was a retrospective case-control study nested within the cohort of children, birth to 18 years of age, undergoing median sternotomy between January 1, 1995 and December 31, 2003. RESULTS: Forty-three cases of mediastinitis were identified. The incidence of mediastinitis was 1.4%. Median patient age at time of inciting sternotomy was 32 days (interquartile range, 5 days-9 months). Twenty three (54%) cases occurred in girls. Median time to onset of infection after surgery was 11 days (range, 4-34 days). Overall Gram-positive organisms were present in 29 (67%) cases, and Gram-negative organisms were present in 13 (30%) cases. The organisms most commonly isolated from mediastinal culture were Staphylococcus aureus (46%), coagulase-negative staphylococci (17%) and Pseudomonas aeruginosa (17%). The rate of concurrent bacteremia was 53% (95% confidence interval, 38-69%). In multivariable analysis, delayed sternal closure was an independent risk factor for the development of Gram-negative mediastinitis (odds ratio, 9.3; 95% confidence interval, 1.5-56.8; P = 0.016). CONCLUSIONS: Although Gram-positive organisms were the most common cause of infection, Gram negative organisms accounted for one-third of all isolates. More than one-half of patients with mediastinitis had concurrent bacteremia. Delayed sternal closure was an independent risk factor for Gram-negative mediastinitis. PMID- 15818291 TI - Comparison of incidence of invasive Streptococcus pneumoniae disease among children before and after introduction of conjugated pneumococcal vaccine. AB - OBJECTIVES: The objective was to determine whether there has been a significant decrease in the incidence of invasive Streptococcus pneumoniae disease in the Louisville, KY, area since heptavalent pneumococcal conjugate vaccine was introduced in the winter of 1999-2000. A secondary objective was to collect demographic data regarding invasive S. pneumoniae disease in the Louisville, KY, area during the 1997-2002 period. METHODS: Data on cases of invasive S. pneumoniae disease (defined by a positive culture for S. pneumoniae from the blood, pleural fluid or cerebrospinal fluid) were collected from the microbiology laboratories and medical records departments of all hospitals that treat children in the Louisville area, for 1997-2002. Trends in case rates for all cases of invasive S. pneumoniae disease from 1999 to 2002 were observed, and demographic and other factors regarding this illness were recorded for each patient. RESULTS: The trend for rates of invasive pneumococcal disease showed a significant decrease during the study period for all ages combined (P < 0.001), for children younger than 2 years of age (P = 0.002) and for children 2 to 5 years of age (P = 0.002). The mean age for children was 2.22 years, with a male/female ratio of 1.8:1. The most common final diagnoses for the patients were bacteremia without focus, pneumonia and meningitis. Forty-eight percent of the organisms were resistant to penicillin. There was a significantly higher rate of resistance to penicillin among S. pneumoniae strains that caused cerebrospinal fluid infection than among strains that caused non-cerebrospinal fluid infection (P < 0.001). The case fatality rate was 2.6%. CONCLUSION: Case rates for invasive S. pneumoniae disease among children decreased significantly in the 2-year period after introduction of the heptavalent S. pneumonia protein conjugate vaccine into this community. PMID- 15818292 TI - Epidemiology of bloodstream infections in the first year after pediatric lung transplantation. AB - BACKGROUND: Substantial morbidity and mortality rates are associated with infections in the first year after pediatric lung transplantation. To understand better the clinical significance of bloodstream infections (BSIs), we evaluated systematically the epidemiologic features of BSIs in the first year after transplantation. METHODS: A retrospective case-cohort study of pediatric primary lung transplant recipients was performed. The frequency of BSIs and the organisms isolated were determined through medical and laboratory record review. We assessed variations in causative organisms and rates of BSIs in 3 time periods after transplantation, ie, early (0-30 days), intermediate (31-90 days) and late (91-365 days). RESULTS: Between July 1990 and November 2000, 190 pediatric patients received primary lung transplants. Twenty-six percent (49 of 190) of recipients had at least 1 BSI. The most commonly isolated organisms were coagulase-negative Staphylococcus (n = 25, 28.4%), Pseudomonas aeruginosa (n = 14, 16.0%) and Candida spp. (n = 9, 10.2%). The overall rate of BSIs was 2.1 episodes per 1000 catheter-days. The highest rate of BSIs occurred in the early period, compared with the intermediate and late periods (5.5, 1.3 and 1.6 episodes per 1000 catheter-days, respectively; P = 0.21). Early BSIs were associated with death in the first year after transplantation (relative risk, 3.9; 95% confidence interval, 1.6-9.4; P = 0.002). CONCLUSIONS: BSIs occur frequently after primary pediatric lung transplantation, with the highest rate being in the first 30 days after transplantation. Early BSIs are associated with death in the first year after transplantation. PMID- 15818293 TI - Predictors of unfavorable outcomes in enterovirus 71-related cardiopulmonary failure in children. AB - BACKGROUND: Enterovirus 71 (EV71) can sometimes cause fatal or disabling diseases in children; therefore EV71-infected children with cardiopulmonary failure were investigated at Chang Gung Children's Hospital to discover the prognostic predictors. METHODS: We investigated 27 EV71-infected children with cardiopulmonary failure from May 2000 to September 2001 and analyzed their clinical data to find predictors associated with unfavorable outcomes of deaths or ventilator dependence. RESULTS: Of the 27 patients, 8 (30%) died and 10 (37%) were ventilator-dependent. Troponin I levels correlated most strongly with fatality, with 5 of the 6 children with troponin I levels >40 ng/ml dying (P = 0.001). Other factors correlated with fatality were cerebrospinal fluid white blood cell count > or =100/microL (P = 0.002) and initial systolic pressure < or =100 mm Hg (P = 0.05). Of the 19 survivors, 10 (53%) were left with central hypoventilation, dysphagia and/or limb weakness plus atrophy. The factors associated with ventilator dependence included higher inotrope equivalent (P < 0.001), duration of hypotension > or =40 hours, initial blood systolic pressure < or =100 mm Hg, positive EV71 isolation and age > or =12 months. CONCLUSIONS: Poor prognostic factors were related to cardiovascular and neurologic damage; therefore physicians may consider advanced cardiovascular support for EV71 infected children with cardiopulmonary failure. PMID- 15818295 TI - Lower respiratory tract infections among american Indian and Alaska Native children and the general population of U.S. Children. AB - BACKGROUND AND OBJECTIVE: Lower respiratory tract infections (LRTIs) cause substantial childhood morbidity. This study characterizes and compares LRTI associated morbidity among American Indian/Alaska Native (AI/AN) children and the general population of U.S. children. METHODS: Hospitalization and outpatient records with a diagnosis indicating LRTIs were evaluated for children aged younger than 5 years during 1990-2001. RESULTS: For 1999-2001, the LRTI associated hospitalization rate was significantly higher for AI/AN children than for U.S. children (116.1 versus 63.2/1000, respectively), with the disparity being greater for infants than for 1- to 4-year-old children. Also the rate of LRTI-associated outpatient visits among AI/AN infants was higher than that for all U.S. infants (737.7 versus 207.2/1000, respectively). LRTI hospitalization and outpatient visit rates were highest in the Alaska and Southwest Indian Health Service regions. During 1990-2001, the LRTI hospitalization rate among AI/AN infants in the Alaska region and among the general U.S. infant population increased. Bronchiolitis-associated hospitalization rates increased for AI/AN and U.S. infants, whereas the pneumonia-associated hospitalization rate decreased among AI/AN infants and remained stable among U.S. infants. CONCLUSIONS: LRTIs continue to be an important cause of morbidity in children, especially among AI/AN infants in the Alaska and Southwest regions. Strategies to reduce LRTI hospitalizations and outpatient visits are warranted for all infants, but the greatest potential impact would be among AI/AN infants. PMID- 15818294 TI - Simple clinical criteria to identify sepsis or pneumonia in neonates in the community needing treatment or referral. AB - BACKGROUND: Sepsis, meningitis and pneumonia annually kill 1.1 million neonates in developing countries; most deaths occur at home. OBJECTIVES: To develop simple clinical criteria, enabling health workers in communities to identify neonates with potentially fatal sepsis; and to identify the danger signs alerting mothers to seek care. METHODS: In a field trial in 39 villages in Gadchiroli, India, trained health workers visited all neonates at home 8 times during the first 28 days of life, recording signs and outcome without interventions during 1995-1996 and with home-based management of sick neonates during 1996-1999. An independent neonatologist assigned the cause of death. We use the term "sepsis" to include sepsis, meningitis and pneumonia. We evaluated 31 signs as predictors of 43 sepsis deaths among 3567 neonates. We also evaluated mothers' observations as the danger signs to seek care. RESULTS: Simultaneous presence of any 2 of 7 signs (reduced or stopped sucking; weak or no cry; limbs becoming limp; vomiting or abdominal distension; baby cold to touch; severe chest indrawing; umbilical infection) predicted sepsis death with sensitivity 100%, specificity 92%, positive predictive value 27.2% and negative predictive value 100% in the nonintervention period. The criteria identified 10.6% of the neonates in the community as suspected sepsis, at a mean of 5.4 days before death. The criteria remained valid in the postintervention period. Any 1 of the 5 maternally observed danger signs (reduced sucking, drowsy or unconscious, baby cold to touch, fast breathing and chest indrawing) gave 100% sensitivity and identified 23.9% neonates for seeking care. CONCLUSION: These criteria identify neonates in the community who are at risk for dying of infection with excellent sensitivity, specificity and negative predictive value but a moderate positive predictive value. They can be used by health workers to select sick neonates for treatment or referral. One potentially fatal case would be treated per 4 presumptive cases treated. PMID- 15818296 TI - Growth of human immunodeficiency virus-infected children receiving highly active antiretroviral therapy. AB - BACKGROUND: Weight and height growth of HIV-infected children tends to lag behind that of uninfected children of similar age. Previous reports of the effect of highly active antiretroviral therapy (HAART) on the growth of HIV-infected children have been contradictory. METHODS: Age- and gender-adjusted height and weight z scores were studied for 192 HIV-infected children, 4 months to 17 years of age, who had been treated with antiretroviral therapy for at least 16 weeks. These children, in clinically and immunologically stable condition, were enrolled into one of 4 HAART regimens and evaluated for 96 weeks. RESULTS: At baseline, these HIV-infected children were significantly shorter than uninfected children (mean z score, -0.57; 95% confidence interval, -0.73 to -0.41; P < 0.001). Children with greater viral loads at baseline were significantly shorter and lighter than children with smaller viral loads (both P < 0.001). Administration of HAART led to an increase in mean weight z scores to normal values (mean z score increase, from -0.16 to >0) by week 48 and an increase in mean height z scores of 72% toward normal values (mean z score increase, from -0.57 to -0.16) by week 96. Younger children gained height more rapidly (P < 0.001), and children with greater baseline viral loads gained weight more rapidly (P < 0.001). There was no evidence of differential height or weight changes in 48 weeks between children with different degrees of virologic control. CONCLUSIONS: HAART improved the average weight gain of HIV-infected children from subnormal to normal after 1 year and improved average height growth to nearly normal after 2 years. PMID- 15818297 TI - Pediatric aspergillosis: disease and treatment differences in children. AB - Invasive aspergillosis is an increasing problem, yet there are few specific data on pediatric aspergillosis. The underlying patient diseases and treatments differ in children and adults, and there is a heterogeneity of invasive aspergillosis that extends to children. Although new diagnostic tools hold great promise for adult patients, it appears that they have limited usefulness in children without the appropriate testing. Antifungal management of pediatric aspergillosis also requires unique dosing schemes that are not used in adult patients. PMID- 15818298 TI - Actinomycosis presenting as osteomyelitis in the pediatric population. AB - Osteomyelitis attributable to Actinomyces often results in recurrent disease and inconsistent responses to antimicrobial agents. We present data for 4 patients and a review of the 15 previously described pediatric cases of actinomycosis presenting as osteomyelitis. Fourteen cases involved the mandible and 5 cases involved other sites. All mandibular cases required at least 1 debridement, with 4 of the cases requiring multiple debridements. PMID- 15818299 TI - A prion primer: transmissible spongiform encephalopathies. PMID- 15818300 TI - Clinical sin nombre hantaviral infections in children. PMID- 15818301 TI - Disseminated Scedosporium prolificans infection and survival of a child with acute lymphoblastic leukemia. AB - Scedosporium prolificans is a saprophytic fungus responsible for an increasing number of infections among immunocompromised hosts. Historically, disseminated infection with this organism has resulted in death. We report on a pediatric patient who developed overwhelming S. prolificans sepsis after induction chemotherapy for acute lymphoblastic leukemia. She is well 18 months after the diagnosis of fungal sepsis and continues to receive chemotherapy for leukemia, which remains in remission. PMID- 15818302 TI - Congenital and neonatal measles during an epidemic in Sao Paulo, Brazil in 1997. AB - During a measles outbreak, 2 mothers with measles gave birth at University Hospital in Sao Paulo City, Brazil. Blood, saliva and urine were collected from the mothers and newborns. Measles virus genome and IgM antibodies against measles were detected. In 1 infant, measles virus genome persisted in peripheral blood mononuclear cells for 157 days after birth. PMID- 15818303 TI - Neonatal suppurative submandibular sialadenitis. AB - Neonatal sialadenitis of the submandibular gland caused by Staphylococcus aureus is described in 2 full term newborns without any risk factors. Fifteen other newborns with isolated submandibular sialadenitis have been reported in the English language literature. Sialadenitis was caused by S. aureus in 87% and occurred primarily in premature gavage-fed neonates (87%). PMID- 15818304 TI - Breast milk as a source of late onset neonatal sepsis. AB - Neonatal sepsis can be a life-threatening complication in preterm neonates. We present the clinical course of 3 preterm neonates, 1 with recurrent sepsis and 2 with late onset sepsis attributed to ingestion of breast milk containing pathogenic organisms. Breast milk should be considered as a potential source of infection in neonates with recurrent infections or when infections occur simultaneously in siblings. PMID- 15818305 TI - CD8 T lymphocytes do not express cytotoxic proteins in coronary artery aneurysms in acute Kawasaki disease. AB - Coronary arterial inflammation in acute Kawasaki disease (KD) is characterized by transmural infiltration of CD8 T lymphocytes, suggesting that CD8 T lymphocyte cytotoxic activity may be important in the pathogenesis of coronary arterial damage in acute KD. We performed immunohistochemistry for the cytotoxic proteins perforin and granzyme B on paraffin-embedded, formalin-fixed coronary artery aneurysm tissue from 6 children who died in the acute stage of KD. Neither perforin nor granzyme B was detected in the KD coronary aneurysm wall. We speculate that the etiologic agent of KD interferes with expression of these cytotoxic proteins by CD8 T lymphocytes, prolonging inflammation in the arterial wall and leading to coronary artery aneurysm formation. PMID- 15818306 TI - Suppurative intracranial complications of sinusitis in previously healthy children. AB - In this retrospective case series, we describe the clinical features and microbiology of 16 previously healthy children with suppurative intracranial complications of sinusitis. Most cases were male adolescents who presented with vomiting in addition to typical sinusitis symptoms. Neurologic signs were often absent. Streptococcus milleri group organisms, along with anaerobes, were common pathogens. PMID- 15818307 TI - Meningitis in a young infant. PMID- 15818308 TI - What is the safe dose of vitamin A in children with measles? PMID- 15818309 TI - Enfurvirtide safety in human immunodeficiency virus-infected children. PMID- 15818311 TI - Acute aseptic meningitis as the only presenting feature of leptospirosis. PMID- 15818312 TI - Neisseria elongata subsp. nitroreducens endocarditis in a seven-year-old boy. PMID- 15818313 TI - Lopinavir/r versus nelfinavir as salvage therapy. PMID- 15818314 TI - A uniform clinical trial registration policy for journals of kidney diseases, dialysis, and transplantation. PMID- 15818316 TI - Adoptive transfer of primed CD4+ T-lymphocytes induces pattern of chronic allograft nephropathy in a nude rat model. AB - BACKGROUND: Chronic allograft nephropathy (CAN) remains the most common cause of late graft loss in renal transplantation. Presensitized patients have a specifically increased risk to lose their graft. To analyze the immunological factors involved, a new experimental rat model was created with nude athymic LEW.RNU rats as recipients of F344 renal allografts. METHODS: Adoptive transfer of CD4+ T-lymphocytes (2x, 3.5x, or 5 x 10(7) cells) primed against donor skin grafts was performed one week after transplantation. The animals were monitored for renal function, graft infiltrating cells, and the development of donor specific alloantibodies for 20 weeks or until graft loss. RESULTS: Survival of the animals was dose dependent; rats suffered from renal failure with severe albuminuria and developed various lesions typical for CAN including interstitial fibrosis and tubular atrophy. The cell infiltrate in the graft increased with the amount of CD4+ T-cells transferred and consists predominantly of CD4+ T-cells and macrophages/monocytes. More than half of the grafts showed histological signs of glomerulopathy consistent with CAN. 9/12 rats with CAN had antibodies against the donor major histocompatibility complex (MHC)-I and in all rats donor specific anti-glomerular basement membrane (GBM) antibodies were detected. CONCLUSION: Adoptive transfer of primed CD4+ T-cells results in a severe infiltrate of CD4+ cells in the graft and production of anti-MHC and GBM antibodies in this nude rat model. Histological changes are consistent with CAN with frequent glomerular changes. In conclusion, the induction of donor specific alloantibodies by primed CD4+ T-lymphocytes may play an important role in the pathogenesis of CAN. PMID- 15818317 TI - Overexpression of human Bcl-2 in syngeneic rat donor lungs preserves posttransplant function and reduces intragraft caspase activity and interleukin 1beta production. AB - BACKGROUND: A significant cause of primary graft failure in lung transplantation is ischemia-reperfusion (I/R). I/R injury generates proinflammatory cytokines, such as interleukin (IL)-1beta, and activates the caspase-mediated pathways of alveolar epithelial apoptosis. The authors investigated whether gene transfer of the human antiapoptotic protein Bcl-2 by means of intratracheal adenoviral administration would preserve posttransplant lung function and reduce intragraft activated caspase activity and IL-1beta production in syngeneic rat donor lung grafts. METHODS: First, 1.0 x 10(9) plaque-forming units of AdvBcl-2 in phosphate buffered saline (PBS), AdvNull empty vector in PBS, or PBS alone was administered intratracheally to ACI (RT1(a)) rats. Then, the left lungs were procured after 24 hr of in vivo incubation and orthotopically transplanted after 1 hr of cold ischemia into syngeneic recipients. After 2 hr of reperfusion, peak inspiratory pressures (PIP) and donor pulmonary vein PaO(2) was measured in all grafts; grafts were then excised and protein extracts were analyzed by enzyme-linked immunosorbent assay (ELISA) and activated caspase-3 and caspase-9 assays. RESULTS: Human Bcl-2 transgene overexpression in donor lung grafts was demonstrated by ELISA of tissue homogenates. Pretreatment of donor lungs with AdvBcl-2 resulted in reduced PIP and increased lung isograft pulmonary vein PaO(2) compared with AdvNull or PBS-alone treated controls. In addition, AdvBcl-2 pretreatment led to diminished cytochrome c release into cytosolic extracts and reduced intragraft IL-1beta production and inhibited intragraft caspase-3 and caspase-9 activity. CONCLUSIONS: Adenoviral overexpression of human Bcl-2 protein limits I/R injury in rat lung isografts. These data suggest that the use of Bcl-2 gene transfer to human lung donors may reduce the oxidative stress of pulmonary grafts after transplantation in clinical lung transplantation. PMID- 15818318 TI - Assessment of pancreatic islet mass after islet transplantation using in vivo bioluminescence imaging. AB - BACKGROUND: Pancreatic islet transplantation is an emerging therapy for type 1 diabetes, but it is difficult to assess islets after transplantation and thus to design interventions to improve islet survival. METHODS: To image and quantify islets, the authors transplanted luciferase-expressing murine or human islets (by adenovirus-mediated gene transfer) into the liver or beneath the renal capsule of immunodeficient mice and quantified the in vivo bioluminescence imaging (BLI) of mice using a cooled charge-coupled device camera and digital photon-counting image analysis. To account for variables that are independent of islet mass such as transplant site, animal positioning, and wound healing, the BLI of transplanted islets was calibrated against measurement of luminescence of an implanted bead emitting a constant light intensity. RESULTS: BLI of mice bearing islet transplants was seen in the expected anatomic location, was stable for more than 8 weeks after transplantation, and correlated with the number of islets transplanted into the liver or kidney. BLI of the luminescent bead and of transplanted islets in the kidney was approximately four times greater than when transplanted in the liver, indicating that photon emission is dependent on optical absorption of generated light and thus light source location. CONCLUSION: In vivo BLI allows for quantitative, serial measurements of pancreatic islet mass after transplantation and should be useful in assessing interventions to sustain or increase islet survival of transplanted islets. PMID- 15818319 TI - Lack of cross-species transmission of porcine endogenous retrovirus in pig-to baboon xenotransplantation with sustained depletion of anti-alphagal antibodies. AB - BACKGROUND: Nonhuman primates are potential permissive animals for studying the risk of in vivo infection with porcine endogenous retrovirus (PERV). Anti alphaGal natural antibodies are considered one of the barriers for preventing PERV infection, and it has been postulated that reduction of these antibodies could increase the risk of this infection. The aim of this study was to investigate the role of GAS 914, which depletes anti-alphaGal antibodies, in the potential in vivo transfer of PERV after pig-to-baboon organ xenotransplantation. METHODS: Twenty-seven baboons underwent xenotransplantation with hDAF or hMCP/hDAF transgenic pig organs, including heterotopic heart (n = 14) and kidney (n = 13) transplants. All of them received GAS 914 along with different immunosuppression protocols. PERV sequences were investigated by reverse transcriptase polymerase chain reaction and by polymerase chain reaction assays in samples obtained at autopsy. The presence of PERV-specific antibodies and/or pig xenomicrochimerism was also evaluated. RESULTS: PERV RNA was not detected in any baboon plasma sample. In addition, all plasma samples were negative for PERV antibodies. However, PERV DNA sequences were detected in peripheral blood mononuclear cells from 6 of 14 (43%) animals investigated. Porcine mitochondrial DNA was also found in all of these positive samples and in six of the eight (75%) samples with negative PERV DNA, indicating that the detection of PERV sequences was attributable to xenochimerism. PERV-positive cells as a result of xenochimerism were also found in eight of nine (89%) spleen and lymph node tissue samples tested. CONCLUSIONS: Sustained depletion of anti-alphaGal antibodies does not augment the risk of PERV infection in pig-to-baboon organ transplantation. PMID- 15818320 TI - Intrasplenic transplantation of encapsulated hepatocytes decreases mortality and improves liver functions in fulminant hepatic failure from 90% partial hepatectomy in rats. AB - BACKGROUND: Encapsulated cell therapy might be a promising approach to enable cell transplantation without immunosuppression. This study investigates the viability and hepatic function of hepatocytes encapsulated with alginate/poly-L lysine in vitro and the effect of the intrasplenic transplantation of cultured encapsulated hepatocytes on survival in 90% hepatectomized rats as a preliminary step toward allogeneic hepatocyte transplantation without immunosuppression. MATERIALS AND METHODS: Rat hepatocytes were isolated and encapsulated using alginate/poly-L-lysine. Encapsulated hepatocytes were cultured for 28 days to measure cell viability, liver function, and morphology. Rats were treated with a 90% partial hepatectomy and then immediately underwent the intrasplenic transplantation of the cultured encapsulated hepatocytes, the capsule alone, or the allogeneic hepatocytes without the capsule. The survival rate, liver function, and cell morphology were assessed after transplantation. RESULTS: The cultured encapsulated hepatocytes maintained their viability and showed better metabolic activity than day 0 cultured encapsulated hepatocytes. The encapsulated cells strongly expressed albumin and were positive for periodic acid-Schiff staining. Electron microscopy demonstrated that the microencapsulated hepatocytes retained the structural elements of hepatic cytoplasm and nuclei. Intrasplenic transplantation of the encapsulated hepatocytes increased the survival rate and improved the hepatic function. Encapsulated hepatocytes transplanted into rat spleen survived well and retained their hepatic function. Moreover, dramatic liver regeneration was observed 48 hr after transplantation in the group that received intrasplenic transplantations of encapsulated hepatocytes. CONCLUSIONS: The intrasplenic transplantation of cultured encapsulated hepatocytes improved the survival rate of an acute liver failure rat model induced by a 90% partial hepatectomy. PMID- 15818321 TI - Immunosuppression by the JAK3 inhibitor CP-690,550 delays rejection and significantly prolongs kidney allograft survival in nonhuman primates. AB - BACKGROUND: Janus kinase 3 (JAK3) mediates signal transduction from cytokine receptors using the common chain (gammac). Because mutations in genes encoding gammac or JAK3 result in immunodeficiency, we investigated the potential of a rationally designed inhibitor of JAK3, CP-690,550, to prevent renal allograft rejection in nonhuman primates. METHODS: Life-supporting kidney transplantations were performed between mixed leukocyte reaction-mismatched, ABO blood group matched cynomolgus monkeys. Animals were treated with CP-690,550 (n = 18) or its vehicle (controls, n = 3) and were euthanized at day 90 or earlier if there was allograft rejection. RESULTS: Mean survival time (+/- standard error of mean) in animals treated with CP-690,550 (53 +/- 7 days) was significantly longer than in control animals (7 +/- 1 days, P=0.0003) and was positively correlated with exposure to the drug (r = 0.79, P < 0.01). Four treated animals were euthanized at 90 days with a normal renal function and low-grade rejection at final pathology. Occurrence of rejection was significantly delayed in treated animals (46 +/- 7 days from transplantation vs. 7 +/- 1 days in controls, P = 0.0003). Persistent anemia, polyoma virus-like nephritis (n = 2), and urinary calcium carbonate accretions (n = 3) were seen in animals with high exposure. Natural killer cell and CD4 and CD8 T-cell numbers were significantly reduced in treated animals. Blood glucose, serum lipid levels, and arterial blood pressure were within normal range in treated animals, and no cancers were demonstrated. CONCLUSIONS: CP-690,550 is the first reported JAK3 inhibitor combining efficacy and good tolerability in a preclinical model of allotransplantation in nonhuman primates and thus has interesting potential for immunosuppression in humans. PMID- 15818322 TI - Results of a three-year prospective study of C2 monitoring in long-term renal transplant recipients receiving cyclosporine microemulsion. AB - BACKGROUND: The clinical utility of C(2) monitoring of cyclosporine A microemulsion (CsA-ME) was evaluated in a prospective study of 110 patients more than 12 months posttransplant who demonstrated stable graft function and were receiving CsA-ME and steroids. METHODS: Patients were converted to C(2) monitoring with the CsA-ME dose adjusted to a target C(2) range of 800 to 1,000 ng/mL and followed for 40 + 11 months. RESULTS: At the time of conversion, 57% of patients exceeded the C(2) target, 20% of patients were below the C(2) target, and 23% of patients were within the C(2) target range. The mean dose of CsA-ME decreased from 258 + 88 to 202 + 76 mg/day (P < 0.0001), and the mean C(2) level decreased from 1,052 + 292 ng/mL to 896 + 233 ng/mL (P < 0.0002). There were no episodes of rejection. At last follow-up, 7.3% of patients had developed chronic renal allograft dysfunction. Use of antihypertensive agents decreased significantly (P = 0.0004), and mean total cholesterol decreased from 6.4 + 1.3 to 5.8 + 1.1 (P = 0.0009) after adoption of C(2) monitoring. CONCLUSION: These findings suggest that conversion of maintenance renal transplant recipients from C(0) to C(2) monitoring of CsA-ME offers the clinical benefits of better control of hypertension and dyslipidemia, with effective protection against chronic renal allograft dysfunction. A target C(2) range of 800 to 1,000 ng/mL in maintenance patients receiving CsA-ME dual therapy seems appropriate. PMID- 15818323 TI - Corticosteroid-free immunosuppression with tacrolimus, mycophenolate mofetil, and daclizumab induction in renal transplantation. AB - BACKGROUND: Corticosteroid-free maintenance immunosuppression after organ transplantation eliminates the well-known corticosteroid-related side effects and may help to improve long-term outcome. We investigated whether a corticosteroid free tacrolimus (Tac)/mycophenolate mofetil (MMF) regimen, in combination with daclizumab (Dac) induction therapy, provides adequate immunosuppression after renal transplantation. METHODS: This 6-month, open-label, multicenter, parallel group study involved 538 renal patients randomized (1:1) to a Dac/Tac/MMF regimen (n = 260) or a Tac/MMF/corticosteroids regimen (n = 278) as a control group. RESULTS: Of the patients who completed the study, 88.8% in the Dac/Tac/MMF group were free from corticosteroid therapy at month 6. The incidence of biopsy-proven acute rejection was 16.5% in both treatment groups; the incidence of biopsy proven corticosteroid-resistant acute rejection was 4.3% and 5.0% with Tac/MMF/corticosteroids and Dac/Tac/MMF, respectively (P = NS for both comparisons). Renal function was also similar in both groups: median serum creatinine at month 6 was 125.0 micromol/L (Tac/MMF/corticosteroids) and 131.0 microml/L (Dac/Tac/MMF), P = 0.277. The overall safety profile was similar with both regimens. However, compared with the Tac/MMF/steroid regimen, a significantly reduced incidence of new-onset insulin-dependent diabetes mellitus (5.4% vs. 0.4%, P = 0.003) was found with steroid-free immunosuppression. Moreover, mean total cholesterol concentrations increased from baseline in the Tac/MMF/corticosteroids group by 0.19 mmol/L, whereas in the Dac/Tac/MMF group, levels decreased by 0.19 mmol/L, P = 0.005. CONCLUSIONS: Corticosteroid-free immunosuppression with a Dac/Tac/MMF regimen is as effective at preventing acute rejection after renal transplantation as a standard triple regimen of Tac/MMF/corticosteroids. Furthermore, the safety benefits reported with Dac/Tac/MMF treatment may help improve the long-term outcome for renal-transplant patients. PMID- 15818324 TI - HSP70-hom gene polymorphism in allogeneic hematopoietic stem-cell transplant recipients correlates with the development of acute graft-versus-host disease. AB - BACKGROUND: A number of genetic polymorphisms have been shown to be associated with the outcome after allogeneic hematopoietic stem-cell transplantation (HSCT). In the present study, HSP70-hom polymorphism (+2763 G/A) was analyzed in the patients and donors of allogeneic HSCT in relation to transplantation outcome, susceptibility for generation of severe toxic lesions, and acute (a) graft-versus host disease (GVHD). METHODS: One hundred thirty-three recipients of allogeneic hematopoietic stem cells and 64 haploidentical and matched unrelated donors were investigated. All these individuals were typed for dimorphism within the HSP70 hom gene (+2763 G/A) with the use of amplification refractory mutation system technique. RESULTS.: Patients with the HSP-AA homozygous genotype presented more frequently with grade II to IV toxic lesions (12 of 14 vs. 61 of 105, P = 0.039) and aGVHD (12 of 16 vs. 56 of 114, P = 0.045). Conversely, DRB1*11 was associated with a lower risk of aGVHD manifestation (10 of 31 vs. 58 of 99, P = 0.009). These contrary associations of HSP-AA and DRB1*11 with the risk of aGVHD were confirmed using logistic regression modeling in multivariable analysis (HSP-AA, odds ratio [OR] = 3.833, P = 0.004; DRB1*11, OR = 0.224, P = 0.048). None of donor HSP genotypes or patient-donor incompatibility within HSP alleles was associated with susceptibility to toxic complications or aGVHD. CONCLUSIONS: Polymorphism of the HSP70-hom gene is associated with the development of posttransplant complications. Recipient HSP-AA homozygous genotype is a risk factor for aGVHD. PMID- 15818326 TI - A study of the metabolites of ischemia-reperfusion injury and selected amino acids in the liver using microdialysis during transplantation. AB - BACKGROUND: Preservation and ischemia-reperfusion injury still impact the outcome of orthotopic liver transplantation. The authors used microdialysis with a view to monitoring its effect on graft function. METHODS: A microdialysis catheter was inserted into the graft immediately after reperfusion and perfused with an isotonic solution for 48 hr. Metabolites of the ischemia-reperfusion injury and selected amino acids were studied. There were 18 patients, with a median age of 52 years (range, 38-62 years), 8 of whom were men. Lactate, pyruvate, glycerol, and glucose levels were measured. In addition, alanine, arginine, citrulline, gamma-aminobutyric acid (GABA), glutamate, glutamine, glycine, and taurine were determined. RESULTS: All grafts functioned well. High lactate, pyruvate, and glycerol levels were observed in the immediate postoperative period. These showed a significant rapid decrease and stabilized to baseline levels. Alanine, glutamate, GABA, and taurine levels declined significantly to baseline values. Arginine levels were low immediately postreperfusion and then increased, reaching significantly higher values beyond 19 hr. CONCLUSIONS: These data may represent "normal" changes seen in the immediate posttransplant period because all grafts functioned well. Two important metabolic fates of arginine in the liver are in the detoxification of ammonia by means of the urea cycle, and in the synthesis of nitric oxide (NO). Low extracellular arginine may reflect influx of the amino acid into hepatocytes, resulting in formation of NO in the presence of inducible NO synthase or conversion to ornithine in the presence of arginase in the urea cycle. As the organ stabilizes, restriction of arginine uptake may give rise to the observed increase in extracellular arginine. PMID- 15818325 TI - Graft-versus-tumor effect against advanced pancreatic cancer after allogeneic reduced-intensity stem cell transplantation. AB - BACKGROUND: The prognosis of advanced pancreatic cancer is extremely poor and therefore a novel treatment strategy is desired. The authors thus started a prospective study of allogeneic reduced-intensity hematopoietic stem cell transplantation (RIST) for patients with advanced pancreatic cancer to evaluate the feasibility and efficacy of this approach for such patients. METHODS: Only patients with pathologically proven pancreatic cancer that was locally advanced or metastatic and not amenable to curative resection were included. The conditioning regimen consisted of gemcitabine, fludarabine, and busulfan. RESULTS: In the first stage of this study, the authors treated seven patients. Treatment-related mortality before day 100 was observed in one patient. The median survival after RIST was 229 days. An objective response on computed tomographic scan was observed in two patients and another had a tumor marker response. Marked tumor shrinkage was observed in one of the remaining patients after donor lymphocyte infusion. These antitumor effects appeared after the effect of the conditioning regimen had disappeared. In addition, some of these responses were associated with an increase in the serum anticarcinoembryonic antigen antibody level. CONCLUSIONS: Pancreatic cancer appeared to be sensitive to a graft-versus-tumor effect; therefore, a larger clinical study with a refined strategy is warranted. PMID- 15818327 TI - The relative importance of cytokine gene polymorphisms in the development of early and late acute rejection and six-month renal allograft pathology. AB - BACKGROUND: Acute rejection episodes and 6-month protocol biopsy acute pathology are highly correlated with long-term outcomes in renal transplant recipients. Recurrent, vascular, and late rejections are particularly deleterious. METHODS: We determined the relative contribution of human leukocyte antigen matching, cytokine genotypes, delayed graft function (DGF), and baseline immunosuppression to the development of acute rejection and allograft pathology in 118 renal transplant recipients. RESULTS: Multivariate logistic regression modeling demonstrated that the adjusted odds ratio and 95% confidence interval for recurrent (> or =2) early rejections (0-3 months) increased linearly for high (H) > intermediate (I) > low (L) interferon-gamma (1.8; 1.1-3.2) and tumor necrosis factor (TNF)alpha (3.0; 1.3-6.9) genotype, whereas every 1 microg/L increase in the cyclosporine A level was protective (0.991; 0.984-0.999). The odds ratio for recurrent late rejections (4-6 months) increased for H > I > L TNFalpha (5.1; 1.8 14.7) genotype and DGF (7.1; 1.6-30.2), whereas H > I/L transforming growth factor-beta1 genotype decreased the relative risk (0.09: 0.02-0.49). Vascular rejection was only predicted by H > I > L TNFalpha phenotype (3.0; 1.2-7.9). The odds ratio for the 6-month Banff Acute Score (6A > or= 4) increased for H > I > L TNFalpha (2.7; 1.1-6.7) and interleukin-10 (3.4; 1.2-6.2) genotype, and DGF (3.4; 1.1-11.5). Treatment of early subclinical rejection decreased the relative risk (0.20; 0.07-0.62). CONCLUSIONS: High transforming growth factor-beta1 producer phenotype seems to be protective against acute inflammation, whereas H and I interferon-gamma, TNFalpha, and interleukin-10 producer genotypes correlate with adverse outcomes. Cytokine genotyping identifies individuals who may benefit from more intensive surveillance and treatment posttransplant. PMID- 15818328 TI - Measurement of apoptosis of intact human islets by confocal optical sectioning and stereologic analysis of YO-PRO-1-stained islets. AB - Apoptosis is an established pathway for islet cell demise. Current protocols for assessment of islet cell apoptosis are time-consuming (as with terminal deoxynucleotide transferase-mediated dUTP nick-end labeling reaction) and involve disruption of the islet architecture (as with flow cytometry) or destruction of cell integrity (as with enzyme-linked immunosorbent assay). The membranes of apoptotic cells, but not those of live cells, are permeant to the DNA-intercalant dye YO-PRO-1. We report a novel methodology for the rapid quantification of apoptosis of human islets: confocal laser optical sectioning and stereologic analysis of intact human islets stained with YO-PRO-1 and Hoechst 33342. The advantages include (1) rapid quantification of apoptosis without disrupting islet architecture and (2) identification of significant heterogeneity in the extent of apoptosis among islets from the same isolate. Confocal laser scanning microscopy microscopic imaging of YO-PRO-1-stained islets may advance investigation of islet cell apoptosis and help develop islet parameters predictive of posttransplant function. PMID- 15818329 TI - The immunological effects of extracorporeal photopheresis unraveled: induction of tolerogenic dendritic cells in vitro and regulatory T cells in vivo. AB - Extracorporeal photopheresis (ECP) may represent an alternative to immunosuppression, as a means of reducing rejection after thoracic organ transplantation. The mechanism by which ECP exerts its protective effects has, until now, remained elusive. We analyzed peripheral blood mononuclear cells of four children with chronic heart and lung transplant rejection, who received ECP in addition to conventional immunosuppressive treatment. The effects of ECP were evaluated at each cycle, comparing blood samples from the same patient collected before and after treatment. In vitro, peripheral blood mononuclear cells treated with ECP undergo apoptosis and are phagocytosed by immature dendritic cells, which, in turn, acquire a tolerogenic phenotype. The frequency of T cells, with a regulatory phenotype and strong suppressive activity, was significantly increased in the blood of ECP-treated patients. The immunomodulatory effects of ECP may be explained by its ability to increase the frequency of regulatory T cells with inhibitory action on transplant immune rejection. PMID- 15818330 TI - Portal vein arterialization in a patient with acute liver failure. PMID- 15818331 TI - Hypereosinophilia in an islet transplant recipient. PMID- 15818332 TI - Successful hepatitis C virus treatment in patients coinfected with HIV after liver transplantation. PMID- 15818333 TI - Complete remission of postransplant lung metastases from hepatocellular carcinoma under therapy with sirolimus and mycophenolate mofetil. PMID- 15818334 TI - Renal transplantation among South Asians: a North American perspective. PMID- 15818335 TI - CD25 saturation rate in heart transplant patients receiving two-dose daclizumab induction. PMID- 15818336 TI - Importance of metastatic site analysis in determining tumor lineage in a lung transplant recipient. PMID- 15818338 TI - Moral absolutes and cultural influences on case management. PMID- 15818339 TI - Twelve years after Eisenberg: do we understand the rationale for CAM use? PMID- 15818340 TI - National Council on Disability seeks to restore ADA. PMID- 15818341 TI - Founder's Syndrome. PMID- 15818343 TI - Think you have a compliance problem? Think again. PMID- 15818344 TI - Medication adherence: factors influencing compliance with prescribed medication plans. PMID- 15818345 TI - The challenge of noncompliance for case managers. PMID- 15818346 TI - Evidence-based assessment and intervention strategies to increase adherence to prescribed medication plans. PMID- 15818347 TI - Understanding and overcoming the challenges of effective case management for patients with chronic wounds. PMID- 15818348 TI - The environmental culture of practice. PMID- 15818349 TI - Advance care planning along the continuum. PMID- 15818352 TI - Reducing risk in infant cardiopulmonary bypass: the use of a miniaturized circuit and a crystalloid prime improves cardiopulmonary function and increases cerebral blood flow. AB - Advances in perfusion strategies have played an important role in improving outcomes following repair of complex congenital heart defects. The influence of cooling strategy, temperature, duration of circulatory arrest, and specific method of cerebral perfusion on neurologic morbidity have been extensively characterized. Similarly, the ability of pharmacologic agents to modulate the post-cardiopulmonary bypass (CPB) inflammatory response has been previously elucidated in both the laboratory and clinical arena. However, modification of the circuit and priming components have received comparably less attention. We recently showed that employment of a miniaturized circuit and a bloodless prime reduce inflammation and have salutary effects on cardiopulmonary function following hypothermic low-flow perfusion (HLF), and that this circuit may also improve cerebral protection following both deep hypothermic circulatory arrest and HLF. The current report, therefore, reviews current strategies utilized to minimize post-CPB inflammation and highlights the empirical evidence from our laboratory demonstrating the beneficial role of a miniaturized extracorporeal circuit in this context. PMID- 15818353 TI - Noninvasive assessment of cardiac output. AB - Improved outcome from shock depends on early detection and correction of circulatory abnormalities. Global cardiac output and oxygen delivery must be adequate and distributed appropriately to meet metabolic demands to prevent the development of multiple organ system dysfunction, prolonged morbidity, and death. Circulatory assessment using standard monitors gives incomplete and sometimes misleading information. This article focuses on the available and emerging technologies that emphasize assessment of blood flow and regional tissue oxygenation. PMID- 15818354 TI - Postoperative management: the role of mixed venous oxygen saturation monitoring. AB - The results of the Norwood operation have improved considerably over the last decade, due in part to improvement in postoperative care. Mixed venous oxygen saturation (MvO2) monitoring has been an important addition to postoperative management. Our use of MvO2 monitoring in Norwood patients has included 96 infants operated from 1996 to the present. This strategy has proven to be technically straightforward and adds information not provided by monitoring systemic saturation alone. MvO2 has a nadir at 6-12 hours after surgery and below a value of 30% is associated with anaerobic metabolism. It identifies patients at risk for early mortality. It also allows evaluation of management of treatment strategies that evolve over time and of specific interventions in individual patients. Optimizing MvO2 constitutes an important goal of postoperative management after the Norwood procedure. PMID- 15818355 TI - Advances in perioperative pacing. AB - Cardiac resynchronization therapy with biventricular pacing has become a significant management tool in adults with heart failure. In children, right rather than just left ventricular failure, is a key problem in the postoperative period. Also, congenital heart defects vary widely in their nature and prognosis. There are now preliminary reports in the literature of the use of multiple temporary pacing sites after congenital heart surgery and acute comparison of the effects of unsynchronized versus synchronized pacing in the postoperative period. These studies support the use of cardiac resynchronization pacing at least as a temporary measure in cases of acute heart failure after surgery in patients with congenital heart disease. PMID- 15818356 TI - Extracorporeal membrane oxygenation for cardiac disease: no longer a mistaken diagnosis. AB - Extracorporeal membrane oxygenation (ECMO) has become a valuable adjunct in caring for infants and children with heart disease. Since the initial reports of ECMO support for cardiac failure in children, the number of centers providing cardiac support and the number of cases of cardiac ECMO have steadily increased. The International Registry for Extracorporeal Life Support Organization has reported survival statistics for cardiac cases in neonates, children, and adults ranging from 33% to 43%. These numbers are similar to the survival from recent reports by Morris (39%) and Chaturvedi (49%). Survival is influenced by ability to be weaned from bypass in the operating room and by residual structural disease and multi-organ system failure but not by cardiac arrest and single ventricle physiology. To improve results in the future, we need to focus on better predicting the need for support and avoiding multi-organ system failure before initiating ECMO. Rapid deployment of ECMO may further improve results for patients who deteriorate suddenly in the intensive care unit. PMID- 15818357 TI - Decision making for mechanical cardiac assist in pediatric cardiac surgery. AB - The practice of pediatric cardiac surgery has evolved to the point where the majority of patients operated on represent the most complex end of the spectrum of congenital heart disease. Given this, the potential role for mechanical cardiac assist will continue to expand. Although extracorporeal membrane oxygenation remains the mainstay of mechanical circulatory assist, the increased use of centrifugal ventricular assist devices is changing the approach to the treatment of acute cardiac failure. A range of newly developed implantable and paracorporeal devices is beginning to make its way into the clinical practice of pediatric cardiac surgery. This article addresses the different types of support available for mechanical cardiac assist and the clinical considerations in selecting the appropriate device. PMID- 15818359 TI - The double switch operation with accent on the Senning component. AB - In congenitally corrected transposition (ccTGA) the most common configuration is atrial situs solitus with left ventricular loop and left transposition of the great arteries (SLL). Less common is ccTGA with atrial siti inversus (IDD). In both configurations there is a high incidence of ventricular septal defect, pulmonary stenosis, or atresia and some anatomic abnormality of the morphologic tricuspid valve (mTV). The morphologic right ventricle (mRV) is the systemic ventricle and prone to premature failure, particularly in the presence of early TV regurgitation, atrial arrhythmias conduction defects, and prior surgical ventricular septal defect closure. With a long experience with the Senning operation and then the arterial switch, it seemed feasible that these could be combined in ccTGA to restore the mLV to the systemic circuit. This was first attempted in 1989 by the author and was successful. Many of the more recently graduated congenital heart surgeons have little or no experience with the inflow switch. For this reason, the author was asked to write this article, accenting the technical details of the inflow switch. The author uses the Senning operation, with those modifications needed to accommodate the differences between the morphologic right atrium, conduction system, and quite frequent discordance between the atrial situs and the position of the apex of the heart, in ccTGA as compared with TGA. PMID- 15818358 TI - The double switch for atrioventricular discordance. AB - Conventional surgery for atrioventricular discordance usually associated with ventricular arterial discordance leaves the morphologic right ventricle in the systemic circulation. Long-term follow-up results with this approach reveal a high incidence of right ventricular failure. The double switch procedure was introduced to restore the morphologic left ventricle to the systemic circulation. This operation is performed in two main ways: the atrial-arterial switch and the atrial switch plus Rastelli procedure. This double switch approach has been successful at least in the medium term in abolishing morphologic right ventricular failure and its associated tricuspid valve regurgitation. In the atrial-arterial switch group, there is an incidence of morphologic left ventricular dysfunction, sometimes associated with neoaortic valve regurgitation, and the minority of cases need aortic valve replacement. The long-term function of the morphologic left ventricle and the aortic valve need careful surveillance in the future. The atrial-Rastelli group of patients has not in the medium term shown evidence of ventricular dysfunction but will require change on a regular basis of their ventricular to pulmonary artery valved conduits. PMID- 15818360 TI - Off pump palliation for hypoplastic left heart syndrome: surgical approach. AB - The ongoing effort to improve the outcomes of patients with hypoplastic left heart syndrome and the awareness about their neurologic function has prompted consideration for a palliative strategy that does not subject these neonates to a major intervention associated with the use of cardiopulmonary bypass with or without a period of circulatory arrest. Ductal stenting and pulmonary artery banding, initially reported over a decade ago, offers an alternative form of initial palliation with minimal intervention during the newborn period while keeping all treatment options available. We describe some of the recent experiences and discuss indications, results, and issues to be addressed to further this approach. PMID- 15818361 TI - Catheter-based interventional palliation for hypoplastic left heart syndrome. AB - Catheter-based interventional palliation of infants with hypoplastic left heart syndrome (HLHS) has been successfully used to stabilize infants awaiting orthotopic heart transplantation. The three critical palliative requirements for HLHS are ductal patentcy, left heart decompression, and the regulation of pulmonary flow. We reviewed our experience in 40 infants with ductal stenting, and 17 of these 40 infants underwent placement of internal pulmonary artery bands. The infants ranged in age from 2 days to 7 months and from 2.1 to 6.6 kg in weight. There was one procedure-related death after balloon septostomy. The duration of palliation was from 3 days to 1 year. Late survival was 75% with long term palliation or transplantation. Experience and anatomy of the RV outflow tract/ductus arteriosus was related to technical complications. Infants with mesoversion (12%) were more likely to have procedural and late complications (65%). Internal pulmonary artery bands provide predictable and effective protection of the distal pulmonary artery bed. Pulmonary artery reconstruction was not required, and there were no late pulmonary artery stenoses. We conclude that interventional catheter-based palliation of HLHS has been proven in principle, and further gains in experience and technology will likely make this desirable in the majority of infants with hypoplastic left heart syndrome. PMID- 15818362 TI - Hybrid procedures in pediatric cardiac surgery. AB - Hybrid pediatric cardiac surgery is an emerging field that combines skills and techniques used by pediatric cardiac surgeons and interventional pediatric cardiologists. This article describes the emerging indications and techniques in hybrid pediatric cardiac surgery and discusses potential future applications. It focuses on peratrial and perventricular septal defect closure, intraoperative stenting, hybrid stage I palliation for hypoplastic left heart syndrome, and percutaneous valve implantation. PMID- 15818363 TI - Septal myectomy for obstructive hypertrophic cardiomyopathy. AB - Septal myectomy effectively relieves left ventricular outflow tract obstruction (LVOTO) and cardiac symptoms in both adults and children with obstructive hypertrophic cardiomyopathy (HCM). Abnormal attachments of the papillary muscles and chordae and other cardiac lesions can be repaired at the same time. Early mortality for isolated septal myectomy in both children and adults is low (0% to 2.5%). Median echocardiographic LVOT gradients at rest on late follow-up have been as low as 0 to 5 mm Hg. Symptomatic improvements after myectomy occurs in the majority; 90% of patients improve by at least one functional class, and most remain improved on late follow-up. Late survivorship compares very favorably with the natural history of nonoperated patients with symptomatic obstructive HCM. These results serve as a basis for comparison with newer nonsurgical alternatives. PMID- 15818364 TI - Interrupted aortic arch: surgical decision making. AB - Interrupted aortic arch (IAA) is defined as the loss of luminal continuity between the ascending and descending aorta. It is associated with a multitude of lesions ranging from isolated ventricular septal defects to complex ones. Although results have improved in the modern era, repair of IAA is associated with a significant mortality and morbidity. In recent years, the move to a one stage repair has become well established, and the optimal technique for aortic repair seems to be partial direct anastomosis with patch augmentation. Left ventricular outflow tract obstruction (LVOTO) continues to be an important factor affecting survival and re-intervention rates after IAA repair. Great variability exists with regard to definition and diagnosis of LVOTO. To guide the decision for left ventricular outflow tract (LVOT) intervention and which type to use, we propose a simple formula based on the baby's weight. We advocate a conservative approach when the LVOT diameter is greater than the baby's weight + 2 mm and a LVOT bypass procedure (Yasui or Norwood) if the LVOT diameter is less than the baby's weight in millimeters. If the LVOT diameter falls in between, no definitive recommendation can be made, and the surgical approach is based on the surgeon's experience and overall philosophy. PMID- 15818365 TI - Valve-sparing aortic root replacement in Marfan syndrome. AB - Marfan syndrome is the most common inherited connective tissue disorder, affecting approximately 1 in 10,000 live births. The cardinal features of Marfan syndrome are the abnormalities of the skeleton (tall stature, arachnodactyly, and joint hyperelasticity), eye (lens subluxation), and aorta (root aneurysm with proclivity toward rupture and dissection). Aortic catastrophe accounts for most of the premature mortality among Marfan patients, a risk that climbs steeply during adolescence and results in death of half of Marfan patients by the age of 40 years. Most of the improvement in life expectancy that has been achieved in Marfan syndrome is attributable to early recognition of aortic root aneurysms and prophylactic replacement with composite grafts (mechanical valve prostheses within Dacron conduits) before rupture or dissection occurs. Despite the excellent early and late results with composite grafts, there has been growing interest in operative procedures that replace the sinuses but preserve the aortic valve leaflets, to avoid anticoagulation and minimize the risk of prosthesis related endocarditis. These procedures are still in evolution and late results are not yet known, but as with mitral repair in the setting of myxomatous disease, valve-sparing procedures in Marfan syndrome have weathered a storm of initial criticism and skepticism and are steadily gaining acceptance. PMID- 15818366 TI - Aortic valve repair. AB - Aortic valve replacement options are limited in children, and all of them have disadvantages. Aortic valve repair techniques have evolved slowly and have not gained wide acceptance; however, large series using a variety of techniques demonstrate that valve repair is possible with excellent early hemodynamics and satisfactory intermediate durability. The results of aortic valve repair at the Children's Hospital of Wisconsin are presented. Simple repairs (blunt valvotomy, commissurotomy, or commissurotomy with leaflet thinning) directed at congenital aortic stenosis resulted in 86% +/- 5% freedom from reintervention at 10 years. Repair of aortic insufficiency with ventricular septal defect (VSD) resulted in 93.3% +/- 6% freedom from reoperation at 10 years. Complex repairs included a combination of techniques and yielded 5-year freedom from reintervention of 83% +/- 7% compared with 73% +/- 11% for patients undergoing aortic valve replacement (P = .62). Aortic valve repair provides an alternative to aortic valve replacement in selected patients. The utility of aortic valve repair and aortic valve replacement must be measured not only in freedom from reintervention but also in regression of left ventricular mass and exercise testing. Improvement in outcome depends on better patient selection and suitable bioprosthetic materials. PMID- 15818367 TI - Surgical therapy for anomalous aortic origin of the coronary arteries. AB - Anomalous aortic origin of the coronary arteries is a rare defect and is associated with an increased risk of cardiac symptoms including sudden death. The most important subtypes are those in which the anomalous coronary artery arises from the wrong sinus as in anomalous left main coronary artery from the right sinus and anomalous right coronary artery from the left sinus. Ischemia can result when the flap-like orifice becomes obstructed, usually during exercise. Indications for surgical intervention are evolving. Currently, the favored approach for repair is an unroofing type of procedure that results in relocation of the coronary artery into the appropriate sinus of Valsalva. PMID- 15818368 TI - An apology for primary repair of tetralogy of Fallot. AB - The first repair of tetralogy of Fallot (TOF) was 50 years ago, so it would seem that the details for optimal management strategies would be clear. Timing of repair and operative repair strategy for TOF are still a source of debate. Varying institutions have published excellent outcomes with a primary repair strategy or a selective staged repair strategy. In this article, the current and historic strategy for repair of TOF at the Hospital for Sick Children in Toronto is delineated along with associated outcomes. Data from our institution indicate a clear survival advantage for primary repair of TOF. PMID- 15818369 TI - Repair of tetralogy of Fallot with absent pulmonary valve using a new approach. AB - Although several techniques for the correction of tetralogy of Fallot (TOF) with absent pulmonary valve have been described, the method of choice is controversial. Symptomatic infants, in particular, have a poor prognosis because of severe central pulmonary artery dilatation and bronchial compression. An alternative technique for primary repair of TOF with absent pulmonary valve syndrome is suggested. Apart from correction of TOF, this approach includes translocation of the pulmonary artery anterior to the aorta and away from the airways. This technique has a potential to reduce or eliminate bronchial compression by the pulmonary artery. Insertion of valve homograft with anterior and/or posterior plication of the pulmonary artery is considered especially in symptomatic newborns and infants. The intermediate-term functional outcomes have been encouraging, with zero mortality events even in the youngest age group of patients and with the disappearance of respiratory symptoms in the majority of them. PMID- 15818370 TI - Surgical management of unbalanced atrioventricular canal defect. AB - Approximately 10% of endocardial cushion defects exhibit unbalance at the atrioventricular inlet. When the atrioventricular valve sits more over one ventricle than the other, the contralateral ventricle is typically hypoplastic. Surgical intervention for unbalanced atrioventricular canal has a much higher morbidity and mortality than for the balanced form of the defect. With unbalanced atrioventricular canal to the right, no universal criteria are in place to choose single versus biventricular repair. In many cases, risk factors have been extrapolated from other lesions with left ventricular hypoplasia. Even if biventricular repair is successful, the reoperation rate is high for this lesion. Little data exist in the literature regarding left unbalanced atrioventricular canal. In general, right ventricular hypoplasia is better tolerated than left ventricular hypoplasia, and biventricular repair is usually possible. If cyanosis or high systemic venous pressure results, the one and one half ventricle repair (biventricular repair with bidirectional Glenn anastomosis) is an option. This article reviews the present understanding of unbalanced atrioventricular canal and discusses diagnostic and surgical strategies for this complex lesion. PMID- 15818371 TI - Late outcomes in patients with surgically treated congenital heart disease. AB - Optimizing late outcomes should be the end result of improvements in medical and surgical care for congenital heart disease (CHD). In addition to mortality, significant morbidities after surgery for CHD need to be considered. These include the need for reintervention, cardiovascular complications, exercise limitations, neurocognitive morbidities, effects on pregnancy, difficulty obtaining insurance, need for chronic medications, and impaired functional status and quality of life. Long-term outcome studies are difficult to perform, and their interpretation is complicated by intervening changes in management. Specific discussion of long-term follow-up of tetralogy of Fallot, D transposition of the great arteries, and hypoplastic left heart syndrome illustrates the myriad management changes over the last three decades, the challenges in predicting outcomes for recent patients, and the need for ongoing initiation of long-term follow-up studies. PMID- 15818372 TI - Evaluation of quality of care for congenital heart disease. AB - There is widespread recognition that surgical outcomes differ by surgeon and institution; however, the definition and measurement of quality in pediatric cardiac surgery is in its infancy. This article discusses the definition of quality, what has been done to define and measure quality of pediatric cardiac surgery, and how to proceed. Descriptions of assessment of quality by evaluating structure, process, and outcome measures; efforts to establish a global congenital heart surgery database; and a comparison of risk-adjusted mortality rates using the Risk Adjustment for Congenital Heart Surgery method are included. PMID- 15818373 TI - Late results and quality of life after pediatric cardiac surgery in Finland: a population-based study of 6,461 patients with follow-up extending up to 45 years. AB - This population-based study characterizes the history and progress of pediatric cardiac surgery in Finland. For the first time in the literature, all defects and procedures are included in an outcome study, reflecting the true effectiveness of operative treatment. All the patients operated on since the beginning of pediatric cardiac surgery in Finland in 1953 and through the end of 1989 were included in this population-based, long-term outcome study. The survival was defined from the Finnish Population Registry Center. The survival rates were compared with those of an age- and sex-matched general population. During the study period, 6,461 patients underwent surgery; 96% of them were traced. A questionnaire was used to study the quality of life. The 45-year survival of patients was 15% less than that of the general population. The survival and the number of necessary operations varied widely with the defect. The patients coped well with their defects when compared with the general population. The education level was similar, and the employment level was higher than expected. Patients were living in a steady relationship as often as the general population, but parenthood was less frequent than expected. The incidence of congenital heart disease among the children of the patients was 2.4%. The overall survival of children operated on for heart defects is fairly good. The patients, especially those with simpler defects, do not have increased risk of death years after successful operation. The majority of patients are feeling well, and their life situation is similar to the general population. PMID- 15818374 TI - The Ross procedure is an ideal aortic valve replacement operation for the teen patient. AB - The Ross procedure is an ideal aortic valve replacement for the teenage patient because the pulmonary autograft is durable, is nonthrombogenic, has excellent hemodynamics, and grows. Since 1992, our center has performed 194 Ross procedures, and 53 of these were in teenagers (10 to 21 years of age). In this group, there have been no perioperative deaths, hospital length of stay was 4 days, and re-operation for autograft failure was only 2% at mean follow-up of 69 months. All patients are in NYHA heart failure class I. Because of its proven efficacy, the Ross operation is our preferred aortic valve replacement for the teenage patient. PMID- 15818375 TI - The Ross procedure is not the procedure of choice for the teenager requiring aortic valve replacement. AB - The Ross procedure was a remarkable technical achievement when it was introduced in 1967. Although the long-term results for the procedure when performed in adults using the technique of intra-aortic subcoronary freehand implantation have been satisfactory, the results for children using the technique of complete aortic root replacement have been less satisfactory. Not only have early outgrowth, calcification, and shrinkage of the pulmonary homograft used to reconstruct the right ventricular outflow tract been a problem but in addition some children have experienced excessive dilation of the neoaortic root as well as neoaortic valve regurgitation. In contrast to the Ross procedure, aortic valve repair and other forms of aortic valve replacement do not exclude the possibility of reoperation in the future using more advanced options such as tissue engineered leaflets or valves. Until more information is available regarding long term performance of the pulmonary root when implanted as a neoaortic root, as well as improved results for reconstruction of the right ventricular outflow tract, the Ross procedure should be used rarely. PMID- 15818376 TI - Modification to the Ross procedure to prevent autograft dilatation. AB - Dilatation of the pulmonary autograft is a problem described following Ross procedure for aortic valve replacement. Patients at risk seem to be those with aortic insufficiency, bicuspid aortic valves, and those with aneurismal ascending aortas. We describe a technique for encasing the pulmonary autograft in a Dacron tube to prevent dilatation in these patients. This technique is reproducible and includes sewing the coronary arteries to all layers of the autograft and Dacron construct. Short-term follow-up shows excellent outcomes with respect to autograft valve function and lack of annular or sinotubular dilatation. This procedure may be useful for extending the Ross procedure to young adults, where autograft growth is no longer needed, to provide a non-dilatable neoaortic root. PMID- 15818377 TI - Surgeons should provide minimally invasive approaches for the treatment of congenital heart disease. AB - The basis for pursuing techniques for less invasive surgery on children with congenital heart disease is to reduce the known long-term morbidities of thoracotomy and sternotomy. In addition, rapid return to normal activities, reduced length of stay, and better pain control may be achieved. Several congenital lesions have been successfully treated with innovative minimally invasive techniques; however, further technique and technology development is required to accomplish repair of the more complex defects safely and effectively. PMID- 15818378 TI - The case against minimally invasive cardiac surgery. AB - Stellar outcomes have been achieved for patent ductus arteriosus (PDA), atrial septal defect (ASD), and ventricular septal defect (VSD) repairs by using the traditional surgical methods. Over the last decade, minimally invasive procedures have been introduced that promise excellent results with an improved cosmetic appearance, shorter rehabilitation period, less pain, and decreased hospital cost. We reviewed various minimally invasive procedures that are used in PDA, ASD, and VSD to assess their safety and efficacy. These techniques use limited approaches to the heart (partial sternotomies, transxiphoid, anterolateral thoracotomy, and mini-thoracotomy) that reduce the surgeons access and control of the cardiac structures. Cannulation sites for the establishment of cardiopulmonary bypass have been altered. Minimally invasive therapies for congenital heart surgery cannot be adopted until evidence-based data has proven them to be equal or better than the traditional procedure. PMID- 15818379 TI - Opinions from the audience response survey at the first joint meeting of the Congenital Heart Surgeons' Society and the European Congenital Heart Surgeons Association. PMID- 15818380 TI - Preclinical full-scale evaluation of dendritic cells transfected with autologous tumor-mRNA for melanoma vaccination. AB - Most cancer vaccines to date have made use of common tumor antigens or allogenic cancer cell lines. The majority of tumor antigens may, however, be unique patient specific antigens. Dendritic cells (DCs) are the most potent antigen-presenting cells known. The present report is a full-scale preclinical evaluation of autologous DCs transfected with autologous tumor-mRNA (tDCs) for vaccination in malignant melanoma. By using autologous tumor-mRNA, we intend to make the DCs present a broad spectrum of tumor-associated antigens relevant to each individual patient. Previously, we have described effective methods for mRNA-transfection into DCs by square-wave electroporation and for generating large numbers of DCs. Here, we demonstrate the ability of tDCs, made under full-scale vaccine conditions, to generate in vitro T-cell responses specific for antigens encoded by the transfected tumor-mRNA. T-cell proliferation assays demonstrated tDC specific responses for all six patients tested. Responses were further studied by IFNgamma ELISPOT and Bioplex cytokine assays (two patients) and by experiments on isolated CD4(+) and CD8(+) T cells, including HLA-blockage (one patient). Moreover, we describe the results of extensive tumor-RNA analysis using Agilent Bioanalyser, a method that we have implemented in the clinical protocol. Based on this preclinical evaluation, a vaccine trial has been started. PMID- 15818382 TI - The potential of oncolytic virus therapy for pancreatic cancer. AB - The objective of this paper was to review a new category of gene therapy using oncolytic viruses for the treatment of pancreatic cancer. The eligibility and feasibility of oncolytic virus therapy as a novel therapeutic agent against pancreatic cancer are discussed as well as basic research for clinical trials, including a historical perspective and the current status of these novel agents. Even combination therapy, such as surgery with radiation and chemotherapy, has not significantly improved the survival rate of pancreatic cancer. Recently, a clinical trial (phase I and II) using an oncolytic adenovirus, ONYX-015, was completed in patients with pancreatic cancer. The phase II trial yielded beneficial results (tumor reduction or stabilization) in about 50% of the patients. A phase I study of the efficacy of oncolytic herpes viruses, G207, OncoVEX GM-CSF, and 1716 against a variety of tumors has been completed, and G207 is in phase II trials for use against brain tumors. In addition, a phase I trial using the herpesvirus showed good tolerance at all dosages. We discuss the basic scientific principles and current results of the above clinical trials with respect to these oncolytic viruses, and then compare the relative advantages and disadvantages of adenoviruses and herpesviruses as oncolytic agents. We also review the published literature on newly developed oncolytic viruses. The concept of oncolytic therapy has been studied for a century. Recent technological developments have made these oncolytic viruses more tumor-specific by exploiting the tumor cell environments. In addition, these viruses have been reported to increase the immunosusceptibility of the tumor cells, and have been designed to express other genes to increase the susceptibility of tumor cells to other therapeutic agents. Oncolytic virus therapy certainly appears to be a feasible treatment for pancreatic cancer. PMID- 15818381 TI - Gene therapy vectors containing CArG elements from the Egr1 gene are activated by neutron irradiation, cisplatin and doxorubicin. AB - Combining gene therapy with radiotherapy and chemotherapy holds potential to increase the efficacy of cancer treatment, while minimizing side effects. We tested the responsiveness of synthetic gene promoters containing CArG elements from the Early Growth Response 1 (Egr1) gene after neutron irradiation, doxorubicin and cisplatin. Human MCF-7 breast adenocarcinoma and U373-MG glioblastoma cells were transfected with plasmids containing CArG promoters controlling the expression of the green fluorescent protein (GFP). Exposing the cells to neutrons, doxorubicin or cisplatin resulted in a significant induction of transgene expression. Therapeutic advantage was demonstrated by replacing the reporter with the herpes simplex virus thymidine kinase (HSVtk), able to convert the prodrug ganciclovir (GCV) into a cytotoxin. A 1.3 Gy neutron dose caused 49% growth inhibition in MCF-7 cells, which increased to 63% in irradiated CArG-HSVtk transfectants treated with GCV. Exposure to 0.5 microM cisplatin or 0.01 microM doxorubicin induced a growth inhibition of 25-30% in MCF-7 cells. In the presence of GCV, this value increased to 65-70% in cells transfected with the CArG promoter constructs driving the expression of HSVtk. These data indicate that combining CArG-mediated HSVtk/GCV suicide gene therapy with radio- and chemotherapy can enhance antitumor toxicity, and validates future in vivo investigations. PMID- 15818383 TI - In vivo targeting of vaccinating tumor cells to antigen-presenting cells by a gene therapy method with adenovirus containing the alpha1,3galactosyltransferase gene. AB - Poor uptake by antigen-presenting cells (APC) is a major reason for low immunogenicity of autologous tumor vaccines. This immunogenicity may be increased by exploiting the natural anti-Gal antibody that is present in humans as approximately 1% of circulating IgG. Anti-Gal binds to alpha-gal epitopes (Galalpha1-3Galbeta1-4GlcNAc-R) on vaccinating tumor cells and opsonizes them for effective uptake by APC. This epitope is synthesized in human tumor cells by transduction with AdalphaGT- a replication deficient adenovirus containing the alpha1,3galactosyltransferase (alpha1,3GT) gene. Protection against tumors by immunization with AdalphaGT-transduced tumor cells was studied in alpha1,3GT knockout (KO) mice, challenged with the highly tumorigenic BL6 melanoma cells. These mice lack alpha-gal epitopes and can produce anti-Gal. Immunization of KO mice with AdalphaGT-transduced BL6 cells protects many of the mice against challenge with live BL6 cells lacking alpha-gal epitopes. Immunization with AdalphaGT transduced autologous tumor cells may serve as adjuvant immunotherapy delivered after completion of standard therapy. This method may complement another gene therapy method in which GM-CSF-secreting vaccinating tumor cells recruit APC to vaccination sites. Anti-Gal-opsonized vaccinating tumor cells will be effectively internalized by GM-CSF recruited APC and transported to draining lymph nodes for processing and presentation of tumor antigens. Alternatively, injection of AdalphaGT directly into solid tumor masses of cancer patients may result in anti-Gal-mediated destruction of the transduced tumor cells in a manner similar to xenograft rejection. The subsequent uptake of anti-Gal-opsonized tumor membranes by APC results in their effective transportation to lymph nodes where processed tumor antigens may elicit a protective antitumor immune response. PMID- 15818384 TI - X-chromosome inactivation: a hypothesis linking ontogeny and phylogeny. AB - In mammals, sex is determined by differential inheritance of a pair of dimorphic chromosomes: the gene-rich X chromosome and the gene-poor Y chromosome. To balance the unequal X-chromosome dosage between the XX female and XY male, mammals have adopted a unique form of dosage compensation in which one of the two X chromosomes is inactivated in the female. This mechanism involves a complex, highly coordinated sequence of events and is a very different strategy from those used by other organisms, such as the fruitfly and the worm. Why did mammals choose an inactivation mechanism when other, perhaps simpler, means could have been used? Recent data offer a compelling link between ontogeny and phylogeny. Here, we propose that X-chromosome inactivation and imprinting might have evolved from an ancient genome-defence mechanism that silences unpaired DNA. PMID- 15818385 TI - Co-evolution of X-chromosome inactivation and imprinting in mammals. AB - Recent studies have revealed mechanistic parallels between imprinted X-chromosome inactivation and autosomal imprinting. We suggest that neither mechanism was present in ancestral egg-laying mammals, and that both arose when the evolution of the placenta exerted selective pressure to imprint growth-related genes. We also propose that non-coding RNAs and histone modifications were adopted for the imprinting of growth suppressors on the X chromosome and on autosomes. This provides a unified hypothesis for the evolution of X-chromosome inactivation and imprinting. PMID- 15818386 TI - Expression of growth factors in the conjunctiva from patients with active trachoma. AB - PURPOSE: The blinding complications of trachoma are associated with progressive conjunctival fibrosis due to excessive accumulation of extracellular matrix (ECM) components. We studied the processes involved in the regulation of fibrosis in trachoma by investigating the expression of the fibrogenic and angiogenic connective tissue growth factor (CTGF) and basic fibroblast growth factor (bFGF), the angiogenic vascular endothelial growth factor (VEGF), the angiogenesis associated endothelial cell marker CD105 (endoglin), and the ECM protein tenascin in the conjunctiva. METHODS: Conjunctival biopsy specimens from six patients with active trachoma, and six control subjects were studied by immunohistochemical techniques using monoclonal and polyclonal antibodies directed against CTGF, bFGF, VEGF, CD105, and tenascin. RESULTS: In the normal conjunctiva, weak immunoreactivity for VEGF was observed in epithelial cells. There was no immunoreactivity for the other antibodies. In all trachoma specimens, immunoreactivity for CTGF and bFGF was localized in monocytes/macrophages, positive for the CD68 marker. Strong immunoreactivity for VEGF was observed in epithelial cells and on vascular endothelial cells. CD105 immunoreactivity was observed on vascular endothelial cells. Immunoreactivity for tenascin was noted in the upper substantia propria. CONCLUSIONS: These findings suggest that macrophages play an active role in conjunctival scarring, upregulated local production of CTGF, bFGF, and VEGF contributes to both fibrous tissue growth and angiogenesis, vascular endothelial cells are activated and are undergoing active angiogenesis, and deposition of tenascin reflect remodelling of the conjunctiva in trachomatous conjunctivitis. PMID- 15818387 TI - Churg-Strauss syndrome in association with proliferative retinopathy. PMID- 15818388 TI - Finding cases of angle-closure glaucoma in clinic setting using a newly developed instrument. AB - PURPOSE: To validate the applicability of a newly developed, noncontact scanning peripheral anterior chamber depth analyzer (SPAC) for screening eyes at the risk of angle-closure glaucoma (ACG). SUBJECTS AND METHODS: All glaucoma patients who visited the University of Yamanashi Hospital from February through May 2003 were enrolled, except those with aphakic eye or pseudophakic eye. Of the 552 enrolled patients, 48 with ACG or narrow angles requiring laser iridotomy (LI) were categorized as patients with high-risk ACG eyes, and those with open angle were categorized as patients with control eyes. In all, 20 patients with ACG or narrow angles requiring prophylactic LI, who were followed up by an independent private ophthalmic clinic, were enrolled for threshold analysis. Nonophthalmologists measured anterior chamber depth and the averaged values of three measurements were employed for analysis. Threshold analysis and discriminant analysis were employed for determining the sensitivity and specificity of SPAC for diagnosing eyes with high-risk ACG. RESULTS: SPAC distinguished well the high-risk ACG eyes from the control eyes, and one of the most useful criteria for screening is as follows: any of the four measured points should exceed 95% confidence interval, and sensitivity and specificity should be 97.6 and 83.5%, respectively. CONCLUSION: SPAC is thought to be useful for detecting eyes at the risk of ACG by nonophthalmologists. PMID- 15818389 TI - Cystoid macular oedema and changes in retinal thickness after phacoemulsification with optical coherence tomography. AB - AIMS: To study the incidence of cystoid macular oedema and changes in retinal thickness after phacoemulsification with optical coherence tomography (OCT). METHODS: In all, 131 eyes of 131 patients were studied at the ophthalmology clinic at Tung Wah Eastern Hospital from September 2001 to October 2002. All the patients had clinical assessment and OCT preoperatively and at weeks 2, 4, and 8 postoperatively. The incidence of postoperative cystoid macular oedema (CMO) was evaluated. The foveal thickness (FT) and central 1 mm retinal thickness (CT) at different time intervals were analysed. RESULTS: Four (3.05%) patients developed CMO after phacoemulsification, which was evident clinically and tomographically. Fluorescein angiogram confirmed leakage in all cases. For other patients, the mean preoperative FT was 189.36 +/- 26.83 microm. The mean FT, were 175.74 +/- 26.79 microm, 180.25 +/- 27.13 microm, 176.58 +/- 26.45 microm at 2 weeks, 4 weeks, and 8 weeks postoperatively, respectively. The preoperative FT was significantly thicker than those in the postoperative period. The same trend was noted for CT. CONCLUSION: OCT is useful for detecting and confirming clinical CMO after cataract surgery; however, its use in detecting subtle changes in retinal thickness is limited by the normal variation in retinal thickness. The measurement of retinal thickness with OCT may also be affected by the status of the lens. PMID- 15818390 TI - Comparison of amplitude of apparent accommodation in pseudophakic eyes with that of normal accommodation in phakic eyes in various age groups. AB - PURPOSE: To compare the amplitude of apparent accommodation in eyes with monofocal intraocular lenses with that of normal accommodation in the phakic fellow eyes in various age groups. PATIENTS AND METHODS: In all, 130 eyes of 130 patients scheduled to undergo cataract surgery, and 130 fellow eyes that had little cataract and good visual acuity of 20/33 or better were studied. The following groups were studied: in their 40s and younger (n = 20) or in their 50s (n = 30), 60s (n = 30), 70s (n = 30), and 80s (n = 20). Using an accommodometer (Kowa HS-9E), the accommodative amplitude was measured at 1 month after surgery. RESULTS: In the patients in their 40s and younger or in their 50s, the amplitude of apparent accommodation was significantly less than that of normal accommodation; no significant difference was observed in the patients in their 60s, 70s, or 80s. The incidence of patients in whom the amplitude of apparent accommodation was more than that of normal accommodation was greater in the patients in their 60s, 70s, and 80s than in the patients in their 40s and younger and in their 50s. CONCLUSION: The amplitude of apparent accommodation is virtually equivalent to that of normal accommodation in patients older than 60 years of age. PMID- 15818391 TI - Computer image analysis of ultrasound biomicroscopy of primate accommodation. AB - PURPOSE: To assess and correct images of the eye for movements that can confound the evaluation of the presence, direction, and magnitude of intraocular movement of the crystalline lens equator during centrally induced ciliary muscle contraction (accommodation). METHOD: Ultrasound biomicroscopic (UBM) video images of a cynomologus monkey crystalline lens were obtained from an independent source. The images, prior to, during, and following electrical stimulation of the Edinger-Westphal (EW) nucleus were compared for evidence of movement of the crystalline lens equator. Extraocular eye movements were assessed by use of objective computer imaging analysis techniques. RESULTS: Extraocular eye movements were identified and reduced by using objective computer imaging analysis techniques to register and realign the corneal images. Highly significant corrections are required to effect corneal realignment. Analysis of paired and registered images from this data source indicates that any movements of the primate lens equator are not detectable when maximum accommodation was induced by EW stimulation. CONCLUSION: The displacement of the edge of the primate crystalline lens equator during electrically induced contraction of the ciliary muscle is a small displacement phenomenon, only analysable after confounding extraocular movements are removed from the compared images. PMID- 15818392 TI - Combined trabeculotomy and trabeculectomy in advanced primary developmental glaucoma with corneal diameter of 14 mm or more. AB - PURPOSE: To report the safety and efficacy of combined trabeculotomy and trabeculectomy (CTT) in advanced primary developmental glaucoma with corneal diameter 14 mm or more. PATIENTS AND METHODS: A total of 74 (121 eyes) consecutive patients aged 3 days-100 months (median, 10 months) with advanced developmental glaucoma were included. Primary CTT was performed by a single surgeon at a tertiary eye care centre in India over a 13-year period. The main outcome measures were changes in and final levels of intraocular pressure (IOP), corneal clarity, visual acuity, and refractive status. RESULTS: Mean preoperative IOP was 29.4+/-7.6 mmHg (range, 10-56 mmHg) and mean postoperative IOP was 15.5+/ 6.6 mmHg (range, 6-38 mmHg) (P<0.0001) with percentage reduction of 44.5+/-27.1. Kaplan-Meier survival analysis revealed 3-, 6-, 9-, 12-, 24-, 48-, and 72-month success rates of 86.5, 80.5, 78.1, 75.5, 71.1, and 60.5%, respectively. Majority (80.5%) of the patients were myopes with mean spherical equivalent of 5.8+/-4.5D (range, 0.75-22.0D). Using various age-appropriate testing procedures, at the final follow-up visit, 14 patients (41.2%) had age-appropriate normal visual acuity and 20 patients (58.8%) had subnormal visual acuity. Best-spectacle corrected visual acuity of > or =20/60 was achieved in seven patients (26.9%). There were no sight-threatening intraoperative and postoperative complications. CONCLUSION: CTT is safe and effective in patients with advanced primary developmental glaucoma. It offers good IOP control with a low complication rate, with about one-third of patients achieving a visual acuity of 20/60 or better. PMID- 15818393 TI - Preschool vision filtering and amblyopia. PMID- 15818395 TI - The thioredoxin system in retroviral infection and apoptosis. AB - Human thioredoxin (TRX) was first identified in human T-cell leukemia virus type I (HTLV-I)-positive T-cell lines and is associated with the pathophysiology of retroviral infections. TRX is a vital component of the thiol-reducing system and regulates various cellular function (redox regulation). Members of the TRX system regulate apoptosis through a wide variety of mechanisms. A family of thioredoxin dependent peroxidases (peroxiredoxins) protects against apoptosis by scavenging hydrogen peroxide. Thioredoxin 2 is a critical regulator of cytochrome c release and mitochondrial apoptosis; transmembrane thioredoxin-related molecule (TMX) has a protective role in endoplasmic reticulum (ER) stress-induced apoptosis. TRX interacts with apoptosis signal-regulating kinase 1 (ASK1) and is a sensor of oxidative stress. Thioredoxin binding protein-2/vitamin D(3) upregulated protein 1 is a growth suppressor and its expression is suppressed in HTLV-I-transformed cells. Studies of these molecules of the TRX system provide novel insights into the apoptosis associated with retroviral diseases. PMID- 15818396 TI - Human spindle checkpoint kinase Bub1 is cleaved during apoptosis. PMID- 15818397 TI - Apoptosis of human endothelial cells is accompanied by proteolytic processing of latent TGF-beta binding proteins and activation of TGF-beta. AB - Transforming growth factors beta (TGF-betas) are multifunctional cytokines that modulate cell growth, differentiation and apoptosis. Numerous effects initiated by TGF-betas in vitro have been described, but the role of TGF-beta targeting and activation under physiological conditions has gained very little attention and understanding. We report here that apoptosis of human umbilical vein endothelial cells (HUVECs) is accompanied by release of truncated large latent TGF-beta complexes from the pericellular matrix followed by activation of TGF-beta. The activation of TGF-beta during apoptosis was accompanied by enhanced secretion of beta1-LAP protein, and apoptotic HUVECs acquired the capacity to induce the release of latent TGF-beta-binding proteins (LTBPs) from extracellular matrices. Activated TGF-beta, in turn, attenuated apoptotic death of HUVECs. Current results indicate that the activation of TGF-beta accompanies the apoptosis of HUVECs, and may play a protective feedback role against apoptotic cell death. The results suggest a role for TGF-beta as a putative extracellular modulator of apoptosis. PMID- 15818398 TI - Morphogenetic plasticity of adult human pancreatic islets of Langerhans. AB - The aim of this study was to investigate the phenotypic plasticity of pancreatic islets of Langerhans. Quiescent adult human islets were induced to undergo a phenotypic switch to highly proliferative duct-like structures in a process characterized by a loss of expression of islet-specific hormones and transcription factors as well as a temporally related rise in the expression of markers of both duct epithelial and progenitor cells. Short-term treatment of these primitive duct-like structures with the neogenic factor islet neogenesis associated protein (INGAP104-118) induced their reconversion back to islet-like structures in a PI3-kinase-dependent manner. These neoislets resembled freshly isolated human islets with respect to the presence and topological arrangement of the four endocrine cell types, islet gene expression and hormone production, insulin content and glucose-responsive insulin secretion. Our results suggest that adult human islets possess a remarkable degree of morphogenetic plasticity. This novel observation may have important implications for understanding pancreatic carcinogenesis and islet neogenesis. PMID- 15818400 TI - Carcinogenesis, cancer therapy and chemoprevention. AB - Carcinogenesis and cancer therapy are two sides of the same coin, such that the same cytotoxic agent can cause cancer and be used to treat cancer. This review links carcinogenesis, chemoprevention and cancer therapy in one process driven by cytotoxic agents (carcinoagents) that select either for or against cells with oncogenic alterations. By unifying therapy and cancer promotion and by distinguishing nononcogenic and oncogenic mechanisms of resistance, I discuss anticancer- and chemopreventive agent-induced carcinogenesis and tumor progression and, vice versa, carcinogens as anticancer drugs, anticancer drugs as chemopreventive agents and exploiting oncogene-addiction and drug resistance for chemoprevention and cancer therapy. PMID- 15818399 TI - In vivo upregulation of CD95 and CD95L causes synergistic inhibition of angiogenesis by TSP1 peptide and metronomic doxorubicin treatment. AB - Antiangiogenic thrombospondin-1 (TSP1) induces endothelial cell death via a CD95 mediated cascade. We used this signaling pathway, where CD95/Fas is a rate limiting intermediate, as a target to optimize the efficacy of TSP1 active peptide, DI-TSP. Like TSP1, DI-TSP upregulated endothelial CD95L in vivo. To modulate CD95 levels, we chose chemotherapy agent doxorubicin (DXR). DXR caused sustained upregulation of CD95 in the activated endothelium at 1/100 of the maximal tolerated dose. DI-TSP and DXR synergistically induced endothelial apoptosis in vitro, and in vivo, in developing murine vessels. Fas decoy, TSP1 receptor antibody and Pifithrin, a p53 inhibitor, severely decreased apoptosis and restored angiogenesis by DXR-DI-TSP combination, evidencing critical roles of CD95 and TSP1. Combined therapy synergistically blocked neovascularization and progression of the bladder and prostate carcinoma. Such informed design of a complex antiangiogenic therapy based on the rate-limiting molecular targets is a novel concept, which may yield new approaches to cancer treatment. PMID- 15818401 TI - DAP5 and IRES-mediated translation during programmed cell death. PMID- 15818402 TI - Role of poly(ADP-ribose) polymerase activity in imatinib mesylate-induced cell death. AB - Imatinib targets Bcr-Abl, the causative event of chronic myelogenous leukemia (CML), and addresses leukemic cells to growth arrest and cell death. The exact mechanisms responsible for imatinib-induced cell death are still unclear. We investigated the role of poly(ADP-ribose) polymerase (PARP) activity in imatinib induced cell death in Bcr-Abl-positive cells. Imatinib leads to a rapid increase of poly(ADP-ribosyl)ation (PAR) preceding loss of integrity of mitochondrial membrane and DNA fragmentation. The effect of imatinib on PAR can be mimicked by inhibition of phosphatidylinositol 3-kinase (PI3-K) implicating a central role of the PI3-K pathway in Bcr-Abl-mediated inhibition of PAR. Importantly, inhibition of PAR in imatinib-treated cells partially prevented cell death to an extent comparable to that observed after caspase inhibition. Simultaneous blockade of both caspases and PAR revealed additive cytoprotective effects indicating that both pathways function in parallel. In conclusion, our results suggest that in addition to the well-documented caspase-dependent pathway, imatinib also induces a PARP-mediated death process. PMID- 15818403 TI - Redundant cell death mechanisms as relics and backups. AB - Here we review recent observations indicating the existence of redundant cell death mechanisms. We speculate that this redundancy reflects a particular evolutionary history for cellular demise. Autophagic or apoptotic elements might have been added to a primordial death mechanism, initially improving cell dismantling and later acquiring the ability to act themselves as death effectors. The resulting redundancy of cell death mechanisms has pathophysiological implications. PMID- 15818404 TI - Protein kinase CK2 phosphorylates and upregulates Akt/PKB. AB - Treatment of Jurkat cells with specific inhibitors of protein kinase CK2 induces apoptosis. Here we provide evidence that the anti-apoptotic effect of CK2 can be at least partially mediated by upregulation of the Akt/PKB pathway. Such a conclusion is based on the following observations: (1) inhibition of CK2 by cell treatment with two structurally unrelated CK2 inhibitors induces downregulation of Akt/PKB, as judged from decreased phosphorylation of its physiological targets, and immunoprecipitate kinase assay; (2) similar results are observed upon reduction of CK2 catalytic subunit by the RNA-interference technique; (3) Akt/PKB Ser129 is phosphorylated by CK2 in vitro and in vivo; (4) such a phosphorylation of activated Akt/PKB correlates with a further increase in catalytic activity. These data disclose an unanticipated mechanism by which constitutive phosphorylation by CK2 may be required for maximal activation of Akt/PKB. PMID- 15818405 TI - Polycystic kidney disease prevented by transgenic RNA interference. PMID- 15818406 TI - IRES in distress: translational regulation of the inhibitor of apoptosis proteins XIAP and HIAP2 during cell stress. PMID- 15818407 TI - p53, apoptosis and axon-guidance molecules. AB - The p53 tumor-suppressor gene regulates apoptosis through the transcriptional activation of its target genes. The expression of the axon-guidance molecule UNC5B (also designated p53RDL1), which is a receptor for netrin-1, is directly regulated by p53. In the absence of netrin-1, UNC5B mediates p53-dependent apoptosis. Conversely, in the presence of netrin-1, p53-induced apoptosis is inhibited through the signaling pathway activated by the interaction between netrin-1 and UNC5B. A number of other molecules that are involved in axon guidance are inactivated in human cancers and are also regulated by p53. These findings suggest a close link between axon-guidance molecules and tumorigenesis. PMID- 15818408 TI - Apoptosis in SIV infection. AB - Pathogenic human immunodeficiency virus (HIV)/Simian immunodeficiency virus (SIV) infection is associated with increased T-cell apoptosis. In marked contrast to HIV infection in humans and SIV infection in macaques, the SIV infection of natural host species is typically nonpathogenic despite high levels of viral replication. In these nonpathogenic primate models, no observation of T-cell apoptosis was observed, suggesting that either SIV is less capable of directly inducing apoptosis in natural hosts (likely as a result of coevolution/coadaptation with the host) or, alternatively, that the indirect T cell apoptosis plays the key role in determining the HIV-associated T-cell depletion and progression to acquired immune deficiency syndrome (AIDS). Understanding the molecular and cellular mechanisms responsible for the disease free equilibrium in natural hosts for SIV infection, including those determining the absence of high levels of T-cell apoptosis, is likely to provide important clues regarding the mechanisms of AIDS pathogenesis in humans. PMID- 15818409 TI - The voltage-dependent anion channel-1 modulates apoptotic cell death. AB - The role of the voltage-dependent anion channel (VDAC) in cell death was investigated using the expression of native and mutated murine VDAC1 in U-937 cells and VDAC inhibitors. Glutamate 72 in VDAC1, shown previously to bind dicyclohexylcarbodiimide (DCCD), which inhibits hexokinase isoform I (HK-I) binding to mitochondria, was mutated to glutamine. Binding of HK-I to mitochondria expressing E72Q-mVDAC1, as compared to native VDAC1, was decreased by approximately 70% and rendered insensitive to DCCD. HK-I and ruthenium red (RuR) reduced the VDAC1 conductance but not that of E72Q-mVDAC1. Overexpression of native or E72Q-mVDAC1 in U-937 cells induced apoptotic cell death (80%). RuR or overexpression of HK-I prevented this apoptosis in cells expressing native but not E72Q-mVDAC1. Thus, a single amino-acid mutation in VDAC prevented HK-I- or RuR-mediated protection against apoptosis, suggesting the direct VDAC regulation of the mitochondria-mediated apoptotic pathway and that the protective effects of RuR and HK-I rely on their binding to VDAC. PMID- 15818410 TI - NF-kappaB factor c-Rel mediates neuroprotection elicited by mGlu5 receptor agonists against amyloid beta-peptide toxicity. AB - Opposite effects of nuclear factor-kappaB (NF-kappaB) on neuron survival rely on activation of diverse NF-kappaB factors. While p65 is necessary for glutamate induced cell death, c-Rel mediates prosurvival effects of interleukin-1beta. However, it is unknown whether activation of c-Rel-dependent pathways reduces neuron vulnerability to amyloid-beta (Abeta), a peptide implicated in Alzheimer's disease pathogenesis. We show that neuroprotection elicited by activation of metabotropic glutamate receptors type 5 (mGlu5) against Abeta toxicity depends on c-Rel activation. Abeta peptide induced NF-kappaB factors p50 and p65. The mGlu5 agonists activated c-Rel, besides p50 and p65, and the expression of manganese superoxide dismutase (MnSOD) and Bcl-X(L). Targeting c-Rel expression by RNA interference suppressed the induction of both antiapoptotic genes. Targeting c Rel or Bcl-X(L) prevented the prosurvival effect of mGlu5 agonists. Conversely, c Rel overexpression or TAT-Bcl-X(L) addition rescued neurons from Abeta toxicity. These data demonstrate that mGlu5 receptor activation promotes a c-Rel-dependent antiapoptotic pathway responsible for neuroprotection against Abeta peptide. PMID- 15818411 TI - Impaired homeostasis and phenotypic abnormalities in Prdx6-/-mice lens epithelial cells by reactive oxygen species: increased expression and activation of TGFbeta. AB - PRDX6, a member of the peroxiredoxins (PRDXs) family, is a key player in the removal of reactive oxygen species (ROS). Using targeted inactivation of the Prdx6 gene, we present evidence that the corresponding protein offsets the deleterious effects of ROS on lens epithelial cells (LECs) and regulates gene expression by limiting its levels. PRDX6-depleted LECs displayed phenotypic alterations and elevated alpha-smooth muscle actin and betaig-h3 expression (markers for cataractogenesis), indistinguishable from transforming growth factor beta (TGFbeta)-induced changes. Biochemical assays disclosed enhanced levels of ROS, as well as high expression and activation of TGFbeta1 in Prdx6-/- LECs. A CAT assay revealed transcriptional repression of lens epithelium-derived growth factor (LEDGF), HSP27, and alphaB-crystallin promoter activities in these cells. A gel mobility shift assay demonstrated the attenuation of LEDGF binding to heat shock or stress response elements present in these genes. A supply of PRDX6 toPrdx6-/- LECs reversed these changes. Based on the above data, we propose a rheostat role for PRDX6 in regulating gene expression by controlling the ROS level to maintain cellular homeostasis. PMID- 15818412 TI - Apoptosis of HIV-specific CD8+ T cells: an HIV evasion strategy. PMID- 15818413 TI - The eukaryotic initiation factor 4E-binding proteins and apoptosis. PMID- 15818414 TI - Role of cardiolipin on tBid and tBid/Bax synergistic effects on yeast mitochondria. AB - The apoptotic effector Bid regulates cell death at the level of mitochondria. Under its native state, Bid is a soluble cytosolic protein that undergoes proteolysis and yields a 15 kDa-activated form tBid (truncated Bid). tBid translocates to mitochondria and participates in cytochrome c efflux by a still unclear mechanism, some of them at least mediated by Bax. Using mitochondria isolated from wild-type and cardiolipin (CL)-synthase-less yeast strains, we observed that tBid perturbs mitochondrial bioenergetics by inhibiting state-3 respiration and ATP synthesis and that this effect was strictly dependent on the presence of CL. In a second set of experiments, heterologous coexpression of tBid and Bax in wild-type and CL-less yeast strains showed that (i) tBid binding and the subsequent alteration of mitochondrial bioenergetics increased Bax-induced cytochrome c release and (ii) the absence of CL favors Bax effects independently of the presence of t-Bid. These data support recent views suggesting a dual function of CL in mitochondria-dependent apoptosis. PMID- 15818415 TI - Human immunodeficiency virus (HIV)-1 proteins and cytoskeleton: partners in viral life and host cell death. AB - Cytoskeletal components play a major role in the human immunodeficiency virus-1 (HIV-1) infection. A wide variety of molecules belonging to the microfilament system, including actin filaments and actin binding proteins, as well as microtubules have a key role in regulating both cell life and death. Cell shape maintenance, cell polarity and cell movements as well as cytoplasmic trafficking of molecules determining cell fate, including apoptosis, are in fact instructed by the cytoskeleton components. HIV infection and viral particle production seem to be controlled by cytoskeleton as well. Furthermore, HIV-associated apoptosis failure can also be regulated by the actin network function. In fact, HIV protein gp120 is able to induce cytoskeleton-driven polarization, thus sensitizing T cells to CD95/Fas-mediated apoptosis. The microfilament system seems thus to be a sort of cytoplasmic supervisor of the viral particle, the host cell and the bystander cell's very fate. PMID- 15818416 TI - tBid interaction with cardiolipin primarily orchestrates mitochondrial dysfunctions and subsequently activates Bax and Bak. AB - TNFR1/Fas engagement results in the cleavage of cytosolic Bid to truncated Bid (tBid), which translocates to mitochondria. We demonstrate that recombinant tBid induces in vitro immediate destabilization of the mitochondrial bioenergetic homeostasis. These alterations result in mild uncoupling of mitochondrial state-4 respiration, associated with an inhibition the adenosine diphosphate (ADP) stimulated respiration and phosphorylation rate. tBid disruption of mitochondrial homeostasis was inhibited in mitochondria overexpressing Bcl-2 and Bcl-XL. The inhibition of state-3 respiration is mediated by the reorganization of cardiolipin within the mitochondrial membranes, which indirectly affects the activity of the ADP/ATP translocator. Cardiolipin-deficient yeast mitochondria did not exhibit any respiratory inhibition by tBid, proving the absolute requirement for cardiolipin for tBid binding and activity. In contrast, the wild type yeast mitochondria underwent a similar inhibition of ADP-stimulated respiration associated with reduced ATP synthesis. These events suggest that mitochondrial lipids rather than proteins are the key determinants of tBid induced destabilization of mitochondrial bioenergetics. PMID- 15818418 TI - [Risk factors for hepatitis C in hemodialysis and its impact on the waiting list for kidney transplantation]. AB - OBJECTIVE: To determine the prevalence and annual seroconversion of Hepatitis C (HCV) in private hemodialysis centers in Lima, correlating them with the transfusion history and permanence time in hemodialysis (HD), identifying, in addition, the hepatic damage and its repercussion in the waiting list for kidney transplants. MATERIAL AND METHODS: Transversal-type descriptive study taken from terminal chronic renal insufficiency (IRCT) carriers (128) who received HD treatment during December 2000. The hepatic biopsies of 20 HCV carriers were collected in the kidney pre-transplant evaluation. RESULTS: Anti-HCV antibodies were present in 76/128 (59%) of the patients. In 6/128 (4.5%) the infection was mixed with the Hepatitis B (HbsAg) virus. The annual seroconversion was 13% (6/48). The positive serology to the HCV was 56% in those who received from one to three transfusions, 66% in those who received from four to nine and 85% for those who received more than ten. The patients with positive serology averaged 54 months of permanence in HD vs. the seronegative patients with an average of 26 months. Thirteen of the twenty (65%) hepatic biopsies showed chronic lesions. From the histological lesions observed, the Persistent Chronic Hepatitis was the most frequent variety, followed by the Hepatic Cirrhosis and the Active Chronic Hepatitis. CONCLUSIONS: The HCV infection is elevated in the Hemodialysis Units of the country. A high percentage of patients excluded from kidney transplants for having necroinflammatory activity and/or hepatic fibrosis is deduced. PMID- 15818419 TI - [Overwhelming course of HCV disease in patients with hypogammaglobulinemia associated with cellular immunity deficiency]. AB - Knowledge of the physiopathological basis of the fibrogenesis in the hepatopathy by hepatitis C virus (HCV) is critical. We describe the evolution of the infection by HCV after a ten-year follow-up in patients with antibody immunodeficiency (common variable immunodeficiency (n=3) (IDVC), IgG subclasses deficiency (n=2), specific deficiency of antibodies formation (n=1). The patients were treated with a prepared intravenous immunoglobulin that was associated later with an HCV hepatitis outbreak. Five of the six patients had a positive overwhelming course (CRP) for HCV and all have changes in their hepatic biochemistry during the exposure period [ Analine Aminotransferase (ALT) (from 280 to 2720 U/L) and Aspartate Aminotransferase (AST) (from 400 to 2600) U/L)]. In less than one year, two patients with IDVC developed cirrhosis and the other patient with IDVC, an active chronic hepatitis while the other patients cured the infection without the treatment. The patients with IDVC presented lower IgG levels than the patients with antibodies deficiency before the exposure (average: seric IgG = 697 mg/dl and 1480 mg/dl respectively) and had, in addition, lower T CD4+ lymphocytes [average: T CD4+ lymphocytes = 22% (413 x 106 cells/l) and 33% (869 x 106 cells/l) respectively)]. One combination of components of humoral and cellular immunodeficiency could play a role in the accelerated evolutive course of the hepatopathy by HCV in patients with IDVC. PMID- 15818420 TI - [New ultrashort scheme for helicobacter pylori infection eradication using tetracyline, furazolidone and colloidal bismuth subcitrate in dyspeptic patients with or without peptic ulceration in the National Hospital Cayetano Heredia]. AB - BACKGROUND: Helicobacter pylori (Hp) infection has been associated with the presence of duodenal ulcer, gastric ulcer and chronic active gastritis. It is also speculated that Hp may have a major role in gastric cancer development. Due to rising antibiotic resistance, probably lack of compliance and the expense of the currently used antimicrobial regimens, it's important to develop efficacious, short-duration and low cost therapies, especially for the treatment of low-income populations from underdeveloped countries. The goal of the present study is to asses the efficacy of two ultrashort antibiotic schemes against Hp infection. METHODS: Patients with diagnosis of Hp infection, found in antral gastric biopsies, were included. They were randomly assigned to receive one of the following therapeutic schemes: tetracycline 500 mg qid, furazolidone 100 mg qid and colloidal bismuth subcitrate 120 mg qid for 3 days (Scheme I) or 4 days (Scheme II). Patients were instructed to come back for follow-up at least 8 weeks after starting medication. At the control visit, an upper endoscopy was performed and an average of 3 antral biopsies was taken. Biopsies were stained with hematoxylin-eosin for histological assessment and with Warthin-Starry silver staining for Hp diagnosis. A single experienced pathologist read all biopsies. In both, the initial biopsy and the control one, we evaluated: presence of Hp; presence, depth and grade of chronic gastritis; presence and grade of inflammatory activity; presence, grade and extent of mucinous damage; presence of glandular atrophy, intestinal metaplasia and lymphoid follicles. We also evaluated dyspeptic symptoms prior and after the treatment, and the presence of adverse events. RESULTS: 80 patients were enrolled, 2 were excluded because of intense nausea and vomits, 4 patients didn't follow the indications properly and 8 patients couldn't be contacted for the control visit. From the remaining 66 patients, 32 were assigned to Scheme I and 34 to Scheme II, both groups were comparable. Eradication rate was 68.8% (22/32) (CI = 52.1% - 82.7%) for Scheme I and 88.2% (30/34) (CI = 74.9% - 96.2%), significant higher, for Scheme II. There was decrease of dyspeptic symptoms and significant improvement of the histological pattern for both groups, except for presence of chronic gastritis, intestinal metaplasia, glandular atrophy and lymphoid follicles. Hp eradication was associated with significant symptoms decrease, normal endoscopy raising and improvement of all the histological parameters, except for presence of intestinal metaplasia and glandular atrophy. Treatment was well tolerated, 57.6% of the patients reported only mild adverse events, nausea was the most frequent (19.7%) and there was no difference between schemes. CONCLUSIONS: The triple ultrashort duration scheme including tetracycline, furazolidone and bismuth for 4 days is efficacious against Hp, with a high eradication rate (88.2%). The Hp disappearance is followed by improvement in every histological parameter that we evaluated, except for glandular atrophy and intestinal metaplasia; and it's also accompanied by a decrease in dyspeptic symptoms. PMID- 15818421 TI - Radical surgical treatment for cystic neoplasms of the pancreas: personal experience. AB - BACKGROUND: Cystic neoplasms comprise 1-10% of all pancreatic neoplasms. Most authors recommend resection due to the difficulties in differentiating benign from malignant tumors prior to operation. We performed an analysis on the radical surgical handling of patients with cystic neoplasms of the pancreas. PATIENTS AND METHODS: We analyzed data of all patients with pancreatic neoplasms who were treated by potentially curative surgical treatment by a single surgeon (A.D.C.). Postoperative survival and complications were recorded. RESULTS: Between August 1983 and November 2003, the aforementioned surgeon performed 77 pancreatic resections for pancreatic tumors. Nine of those operations were performed for cystic neoplasms of the pancreas. All patients submitted to partial pancreatic resections were discharged in good health. The patient that underwent a total pancreatectomy died nine days after the procedure. At a mean follow up of 44 months, there has been one local recurrence with death. CONCLUSIONS: Cystic tumors of the pancreas accounted for 11.7 % of the neoplasms studied in our series. Resection of those neoplasms attained by distal pancreatectomy is a safe procedure. PD is a risky procedure and should be viewed cautiously. Total pancreatectomy remains as a high-risk procedure and should be avoided. PMID- 15818422 TI - [Gastric Cancer in Early Stage: study of 371 lesions in 340 patients in the E. Rebagliati National, Lima-Peru]. AB - The first concrete description of the Gastric Cancer in Early Stage is attributed to the German, M. Verse in 1903, in his book Die Histogenese der Schleimhautcarcinome, but the first to use the term, was the French, Bayle in 1833, calling it First Stage Gastric Cancer. From 1963 to 2002, 5118 gastric cancers were histologically diagnosed, in our hospital from which 2337 (46%) were resected in 340 of the resected specimens, 371 early gastric cancers (EGC) (15.87%) were found. DISTRIBUTION: For every five periods, the number and proportion of early cancers increases, from 10 (6.8%) in the first five years to 78 (21.0%) in the last one. AGE: The average is: 64 years old for men and 58 for women with a global average of 61 years old. The youngest case was in a woman of 23 years old. The oldest case in men was 93 years old, and in women, 81 years old. SEX: 235 males and 105 females, with a ratio of 2,2:1. MACROSCOPY: elevated type: Type l 70 (18.9%) Ila 66 (17.8%) Ila+Ilc 26 (7.0%) lla+l 1 (0.3%) I+lla 0 (0.0) IIa+III 1 (0.3%) IIa+IIb 1 (0.3%). Global average of the elevated type: 44.6%. SUPERFICIAL TYPE: llb 17 (4.6%), DEPRESSED TYPE: llb+llc 10 (2.7%), llc 127 (34.2%), llc+l 0 (0.0%) llc+lll 43 (11.6%) lll 3 (0.8%) lll+llc 5 (1.3%) llc+lla 1 (0.3%) lla+llb 0 (0.0%). The global average of the depressed type is: 50.9%. LOCATION: Antrum 171 (46.1%) body 138 (37.3%) bottom 46 (12.3%) antrum body 12 (3%) and body-bottom 3 (1%). SIZE: Maximum 90 mm, minimum 1.5 mm, average 24.9 mm; 25 (8%) diminute (microcarcinomas), 58 (21%) small. HISTOLOGY: Differentiated 219 (64%) undifferentiated 121 (36%). Degree of differentiation in men 74% and in women 26%. Degree of differentiation in diminutes 92%, in small 79% and in large 69%. DEPTH: 204(55%) in mucous, 167(45%) in mucous/submucous. In differentiated 182 (63%) in mucous, 108 (37%) in mucous/submucous. GANGLION METASTASIS: In the 187 located in mucous, 13 (3.8%) were positive, in the 153 located in mucous/submucous 30 (8.8%) were positive. Global percentage of positive ganglions: 12.3%. SYNCHRONIC LESIONS: General in synchronic lesions in 54.6% in other types of cancer 12.7%. INTESTINAL METAPLASIA: Present in 65%, of which 50% of them were uncompleted. SYMPTOMS AND SIGNS: Dyspepsia 82.2%, hyperoxia 32.9%, loss of weight 23.3%, non-characteristic pain 23.3%, ulcer pain 20.5%, melena 8.2%, anemia 7.7%, diarrhea and/or constipation 7.4%, vomiting 2.2%, hematemesis 1.9%. BLOOD TYPE: O (63.2%) A (24.7%) B (11.3%) AB (0.6%). GASTRIC SECRETION: This was determined in 70 patients: In 55 with ulcer, none showed achlorhydria 0%, 20 (36%) hypochlorhydria, 20 (36%) normochlorhydria and 15 (28%) hyperacidity. In 16 in an elevated form 13 (87.5%) with achlorhydria, 1 (6.5%) with hypochlorhydria, 1 (6.5%) with normal acidity. Average in ulcerated cases DAB 3.04 +/- 1.25 mEqh and DAM 12.31 +/- 8.00. In elevated cases: DAB 0.89 +/- 0.32 and DAM 2.20 +/- 0.95 mEqh. PMID- 15818423 TI - [Endoscopic treatment of early gastric cancer: from Endoscopic Mucosal Resection (EMR) to Endoscopic Submucosal Dissection (ESD)]. AB - Surgery has been the traditional treatment for early gastric cancer, but it is known that the endoscopic treatment offers the same results with a lower mortality rate. The first endoscopic therapy used was the polypectomy in cases of pediculate tumors. However, in depressed or flat tumors, tissue-destructive methods are used together with sclerosant substances, laser or plasma argon; their disadvantage is that the procedure cannot be properly evaluated since the tissue is destroyed. On the other hand, the Endoscopic Mucous Resection (EMR) techniques, such as strip biopsy, the injecting and cutting technique, the transparent capsule or the varices ligation, does permit the evaluation of the results of the endoscopic treatment. Progresses in EMR are due to the use of the chromoscopy and the submucous injection of various substances (normal and hypertonic saline solution, hyaluronic acid, etc.). The problem of the EMR is local relapse, attributed to the resection in lesions fragments of more than 10 mm. To resolve this problem, the Endoscopic Submucous Dissection (ESD) has been developed, using the endoscopic scalpel or its modifications; such as the endoscopic scalpel with the isolated point (IT-Knife). The last ones require more operating time, better skills and have more complications. The efforts to make a better diagnosis and treatment of these lesions continue day by day, by using the magnification endoscopes, the endoscopes with multiple inflexions, etc. PMID- 15818424 TI - [Xanthogranulomatous cholecystitis: retrospective analysis of 6 cases]. AB - Xanthogranulomatous cholecystitis (CX) is a rare kind of chronic cholecystitis, not yet reported in our media, characterized by the presence of chronic, inflammatory infiltration, formation of granulomas, with fibrosis and severe histiocytic reaction with macrophages rich in foam cells. The object of this study is to establish the clinical, radiological and histopathological pattern of CX, by means of the analysis of 6 cases identified in a retrospective check of 191 medical histories of cholecystectomized patients suffering from anatomopathological diagnosis of chronic cholecystitis, in the Department of Abdomen of the Institute of Neoplastic Diseases, from 1939 to 2004. The clinical presentation was characterized by the presence of a palpable mass on physical examination and weight loss. There were complications in two patients. The ultrasonigraph, tomograph and/or laparotomy scans of the vesicle were similar in appearance to a locally advanced vesicular cancer. In none of the specimens was the coexistence of a vesicular carcinoma identified. The vesicle was dried out in block with adjacent hepatic parenchyma in all cases. The CX can simulate a hepatobiliary malignant neoplasia and require suitable oncological surgical treatment. In cases of vesicular tumors, which can be considered inoperable there is the possibility of being faced with a xanthogranulomatous cholecystitis (CX), a benign condition treatable with surgery. PMID- 15818425 TI - [Malignant acanthosis nigricans: case report and literature review]. AB - We present a senile patient that arrived as an outpatient because of marked pigmentation of his face and lower limbs and later develops ascites and loss weight. Biopsies of the skin showed the presence of Acanthosis Nigricans and we later showed its presence as a paraneoplastic syndrome when we discovered a gastric cancer. We discuss the association between Acanthosis Nigricans and cancer as well as the most common neoplasia that present with this entity. PMID- 15818426 TI - [Transduodenal ampullectomy: an useful alternative for the treatment of Vater's ampulla lesions. Case reports of an unfrecuent surgical technique]. AB - The Vater's ampulla neoplasias are not very frequent lesions. The ampullectomy consists on the total resection of the Vater's ampulla and part of the duodenal wall with the later reconstruction and anastomosis of the common bile duct and the Wirsung's duct to the duodenum. The local resection of the ampulla is an alternative to the duodenopancreatectomy that has to be taken into account, especially in benign lesions as in the case of adenomas of the Vater's ampulla. Here we describe two cases one with adenoma of the Vater's ampulla, with a high degree dysplasia, who first underwent an ampullectomy and finally ended up in a duodenopancreatectomy due to the infiltration of the adenoma into the edge of section and other case of Lymphangioma of Vater's ampulla with no infiltration in the edge of section. PMID- 15818427 TI - Immunohistochemical study of severe acute respiratory syndrome-associated coronavirus in tissue sections of patients. AB - BACKGROUND AND PURPOSE: Severe acute respiratory syndrome (SARS) is a life threatening and highly contagious disease caused by the novel SARS coronavirus (SARS-CoV). Immunohistochemical staining for SARS-CoV in the tissue sections of SARS patients is helpful in investigations of the biologic behavior of this virus in human tissue, and to determine the target cells of this virus in different organs. METHODS: We studied the pathologic specimens from 6 SARS patients by immunohistochemical staining using a specific antibody against SARS-CoV. RESULTS: Positive viral staining was only found in the lung tissue taken from the patients who died in the early stage of the disease, usually less than 10 days after symptom onset. No positive staining was found in the lung tissue specimens collected in the mid-to-late stage of the disease. The SARS-CoV-infected cells had a patchy distribution and tended to be present in the periphery of the lung. Immunohistochemically, these viral-infected cells were located mainly along the alveolar space, had a cuboidal appearance, and were reactive to cytokeratin and surfactant protein C. This suggested that type II pneumocytes are the main target cell of SARS-CoV in the lung. Occasional intra-alveolar macrophages were also weakly reactive to SARS-CoV antibody. In addition to the lung, positive viral staining was also found in the mucosal epithelium of the large intestine in another patient who had the clinical symptom of diarrhea. No evidence of viral staining was found in other organs including kidney, liver, lymph node and spleen. The skeletal muscle specimens of 2 patients who had rhabdomyolysis were also negative for SARS-CoV. CONCLUSIONS: SARS-CoV is mainly present in the cytoplasm of type II pneumocytes and can only be detected in the lung tissue during the early stage of the disease. In the patient who had symptoms of diarrhea, SARS-CoV staining was also identified in the mucosal epithelium of the colon. PMID- 15818428 TI - Accuracy of diabetes diagnosis in health insurance claims data in Taiwan. AB - BACKGROUND AND PURPOSE: There are limited data from Taiwan about the accuracy of National Health Insurance (NHI) claims data. This study assessed the accuracy of NHI claims data for diabetes and its associated factors. METHODS: Insurance claims data for patients with a diagnosis of diabetes were extracted from the records of the Bureau of National Health Insurance, including detailed files of the outpatient, emergency, inpatient and pharmacy treatment records from January 1, 2000 to December 31, 2000. A stratified, 2-staged, probability proportional to size and equal probability method was used to sample 9000 diabetes patients. The accuracy of the diabetes diagnosis was assessed based on patient responses to questionnaire items. Subjects with negative or uncertain questionnaire answers who had hypoglycemic agents in pharmacy claims data were also classified as diabetic. RESULTS: A total of 1350 questionnaires were returned and an accurate diagnosis was verified from data in 1007 (74.6%) of these subjects. Univariate analysis showed that level of accreditation of the hospital, age, gender, follow up department, type of complication, number of outpatient visits, emergent visit, as well as hospitalization were significant factors associated with an accurate diagnosis of diabetes. Multivariate logistic regression analysis revealed that number of outpatient visits, hospitalization, age, and the level of accreditation of the hospital were significant independent factors. The odds ratio of an accurate diagnosis increased with the number of outpatient visits and hospitalization. The probability of accurate diagnosis of diabetes among patients with >/= 4 outpatient visits was 99.16 times greater than that of patients with /= 1 hospitalization was 5.26 times that of patients who had not been hospitalized. CONCLUSIONS: This study found that the accuracy of diabetes diagnosis in NHI claims data in Taiwan was 74.6%. Further attention to the association of inaccurate claims in cases with infrequent outpatient visits, young age and those attending non-accredited hospitals is needed in order to address the efficiency of diagnosis and surveillance of diabetes in Taiwan. PMID- 15818429 TI - Value of the pneumonia severity index in assessment of community-acquired pneumonia. AB - BACKGROUND AND PURPOSE: The value of the Pneumonia Severity Index (PSI) in predicting the mortality of patients with community-acquired pneumonia has not been reported in Taiwan. This study investigated the value of this scoring system in estimating mortality of inpatients with community-acquired pneumonia. METHODS: This was a prospective observational study of 118 inpatients and a retrospective chart review of 115 inpatients with radiographically-confirmed community-acquired pneumonia treated at a tertiary referral medical center in southern Taiwan. Patients were stratified into 5 risk classes according to PSI score. Data on demographic characteristics, comorbidities, baseline clinical and laboratory features, in-hospital mortality and length of hospital stay were analyzed. RESULTS: The mortality rates according to risk classification were 0% for class I and II, 2.5% for class III, 8.2% for class IV, and 31.2% for class V. A significant correlation was found between these risk classes and medical outcome (p < 0.001). The length of hospital stay was significantly associated with risk class, and ranged from 6.3 days for class I patients to 18 days for class V (p < 0.001). CONCLUSION: The PSI provided a useful prediction of medical outcome in patients with community-acquired pneumonia. To decrease unnecessary admission, further prospective studies are needed to determine whether outpatient therapy is appropriate for class I or class II patients with community-acquired pneumonia. PMID- 15818430 TI - The role of bronchoscopic assessment in esophageal cancer - clinical and survival analysis in 153 patients. AB - BACKGROUND AND PURPOSE: The diagnostic value and indications for fiberoptic bronchoscopy in the preoperative assessment of patients with esophageal cancer have not been fully studied. We evaluated the role of fiberoptic bronchoscopic examination in the stage work-up of patients with esophageal cancer and correlated the results with survival time analysis. METHODS: The medical records of 153 patients with an initial diagnosis of esophageal cancer were reviewed. Clinical data, bronchoscopic findings, treatment courses, and survival time of these patients were analyzed. RESULTS: On initial bronchoscopic examinations, distortion/compression of the normal structure and protrusion at the posterior wall of the trachea or bronchus were the most common bronchoscopic findings (35.9%). We stratified patients into 3 subgroups according to bronchoscopic findings of direct invasion, external compression, and negative findings. The symptoms of dyspnea, hoarseness, aspiration and fever were more frequent in patients with direct airway invasion compared with patients with external compression and negative bronchoscopic findings (p < 0.02). Washing and brushing cytology examinations were all negative in patients with external compression of the airway. There was a significant difference of survival time among these 3 groups of patients (direct invasion: 5.6 +/- 0.6 months; external compression: 12.3 +/- 0.9 months; negative findings: 13.3 +/- 1.1 months, p < 0.01). Direct airway invasion and original cancer stage were the most important variables for survival in the multivariate analysis, and the hazard ratio for prognosis was 2.5 (95% confidence interval [CI], 1.1-4.6) and 4.2 (95% CI, 1.5-9.3), respectively. Twelve patients (80%) with tracheoesophageal (TE) fistulae died within 3 months after diagnosis due to aspiration pneumonia and septic shock. CONCLUSIONS: The role of bronchoscopic examination in patients with esophageal cancer for preoperative evaluation resides in its ability to predict airway invasion and its impact on survival. Advanced cancer stage (stage IV) and direct airway invasion (especially TE fistula) were significantly associated with poor prognosis. These results suggest that patients suffering from dyspnea, hoarseness, aspiration and fever, implicating a high probability of airway invasion, are more likely to benefit from bronchoscopic examination and proper management in order to prevent aspiration or complications. PMID- 15818431 TI - Invasive fungal infection in children with persistent febrile neutropenia. AB - BACKGROUND AND PURPOSE: Children with persistent febrile neutropenia (fever > 96 hours) refractory to antibacterial therapy carry a high risk of invasive fungal infection (IFI). However, epidemiologic data in such patient populations is lacking in Taiwan. METHODS: A retrospective study was conducted to identify risk factors and outcome of IFI in children with persistent febrile neutropenia (> 96 hours) hospitalized at National Taiwan University Hospital from January 1, 1999 through December 31, 1999. They were categorized into proven, probable, possible, or no IFI according to host, microbiologic and clinical criteria. RESULTS: A total of 29 episodes (35.4%) of IFI occurred in 82 episodes of persistent febrile neutropenia and included 1 proven, 12 probable and 16 possible cases. Pneumonia was most common (69.0%), followed by disseminated infection (17.2%), paranasal sinusitis (6.9%), and hepatosplenic abscess (6.9%). Multivariate analysis revealed 4 risk factors independently associated with IFI: prolonged neutropenia (> 30 days; odds ratio [OR], 10.4; 95% confidence interval [CI], 1.8-60.5), prolonged steroid therapy (OR 2.0, 95% CI 1.0-10.8), allogeneic bone marrow transplantation (OR 5.3, 95% CI 1.0-28.0), and relapsed malignancy (OR 3.5, 95% CI 1.1-11.1). Patients with IFI carried higher in-hospital mortality than those without (51.7% vs 3.8%, p < 0.001). Among those with IFI, delayed antifungal therapy (persistent fever > 7 days) was associated with higher mortality rate than earlier treatment (73.3% vs 28.6%, p = 0.024) and was an independent adverse prognostic factor. CONCLUSION: IFI was common in patients with persistent febrile neutropenia and delayed antifungal therapy was an independent adverse prognostic factor. PMID- 15818432 TI - Prognostic significance of six-month estimated glomerular filtration rate in cadaveric renal transplantation. AB - BACKGROUND AND PURPOSE: We hypothesized a linear relative-risk model for graft survival of cadaveric renal transplantation, with a 6-month estimated glomerular filtration rate (GFR) employed as the linear parameter quantifying functioning renal mass and the detrimental effects of early chronic allograft nephropathy. METHODS: A retrospective study was conducted in a 15-year series of cadaveric renal transplantations (n = 227) with cyclosporin-based immunosuppression in a single transplant center. The Cockcroft-Gault formula was used for estimation of GFR, with a correction factor of 0.85 used for women. Stepwise Cox's regression analyses were applied to examine the prognostic significance of the 6-month estimated GFR. RESULTS: The Kaplan-Meier 5- and 10-year graft survival rates for this study were 80.67% and 59.43%, respectively. From univariate analysis, recipient gender, acute rejection and 6-month estimated GFR were significantly associated with graft survival (p < 0.05). Acute rejection became less significant (p = 0.0033) than 6-month estimated GFR (p = 0.0001) when 6-month estimated GFR was introduced in the stepwise regression procedures. CONCLUSION: We demonstrated that 6-month estimated GFR is a significant prognostic indicator in cadaveric renal transplantation and is an essential parameter in the regression modeling of long-term graft survival. PMID- 15818433 TI - Prevalence of lower urinary tract symptoms during pregnancy in Taiwan. AB - BACKGROUND AND PURPOSE: Women often complain of symptoms related to urination during their pregnancy, but data are limited on the clinical profile and lower urinary tract symptoms (LUTS) of pregnant women in Taiwan. This study assessed the prevalence and predisposing factors of LUTS in Taiwanese women during pregnancy. METHODS: A cross-sectional survey was designed to collect data on the prevalence of LUTS during pregnancy. Women attending the antenatal clinic of a medical center in central Taiwan were recruited and asked to complete a LUTS questionnaire including 12 questions on symptoms related to urination. RESULTS: 799 normal pregnant women were included in this study. The most common LUTS were nocturia (60.2%) and stress urinary incontinence (SUI; 46.1%), followed by urgency (34.1%), frequency (27.8%), incomplete emptying (26.2%), a bearing-down sensation (23.8%), and voiding difficulty (12.6%). The prevalence of nocturia, SUI, urgency, and frequency generally increased as gestational age advanced. There was a significantly higher prevalence of SUI in multiparous women than in nulliparous women; however, the prevalence of urgency, frequency, incomplete emptying, bearing-down sensation, and straining was significantly higher in nulliparous women than in multiparous women. High parity, high body weight before pregnancy and advanced gestational age were the predisposing factors of SUI during pregnancy. Only 6.5% of the pregnant women studied complained that SUI caused a social or hygienic problem based on the International Continence Society (ICS) criteria. CONCLUSIONS: This study provides a profile of LUTS in Taiwanese women during pregnancy. Nocturia was the most common lower urinary tract symptom. Nocturia, SUI, urgency, and frequency increased as gestational age advanced. Parity was a predisposing factor for LUTS during pregnancy, with multiparous women experiencing significantly more SUI while nulliparous women had higher prevalence of other LUTS. PMID- 15818434 TI - Spontaneous pneumothorax after steroid treatment in a patient with bronchiolitis obliterans organizing pneumonia. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is an uncommon pulmonary disorder which is a clinicopathologic syndrome. The typical symptoms include cough, fever and dyspnea. Corticosteroid therapy is effective. Death from progressive disease is infrequent. We report a case of BOOP in a 41-year-old male non-smoker, proved by pathologic findings. After steroid therapy (prednisolone 30 mg daily) for 10 days, he developed spontaneous pneumothorax of bilateral lung fields. Responses to steroid and chest tube thoracostomy were excellent. BOOP may be complicated with spontaneous pneumothorax which may adversely affect prognosis. BOOP should be included in the differential diagnosis of patients presenting with secondary spontaneous pneumothorax. PMID- 15818435 TI - Hyperemesis gravidarum presenting as jaundice and transient hyperthyroidism complicated with acute pancreatitis. AB - Hyperemesis gravidarum is an extreme form of nausea and vomiting during pregnancy. Its presenting symptoms include vomiting, disturbed nutrition, electrolyte imbalance, ketosis, extreme weight loss, renal and/or liver damage. It is rare for a hyperemesis gravidarum patient to present with jaundice, hyperthyroidism and idiopathic acute pancreatitis during the same hospitalization period. Here, we report such a case. A 25-year-old pregnant woman without underlying liver or thyroid disease was admitted due to jaundice noted for 2 days at 8 weeks of gestational age. Hyperthyroidism symptoms of tachycardia and finger tremor also bothered her. After treatment with parenteral fluid and antithyroid agents, her clinical condition improved. However, an episode of idiopathic pancreatitis occurred after nausea and vomiting subsided. Bowel rest with parenteral fluid and nutrition supplement was given and the increased pancreatic enzyme level gradually subsided. Follow-up liver and thyroid function were normal after gestational age of 26 weeks. She delivered a healthy female baby without low birth body weight at gestational age of 39 weeks. Rapid diagnosis and supportive care are important for the hyperemesis gravidarum patient with the complication of acute pancreatitis. PMID- 15818436 TI - Novel PHEX gene mutations in two Taiwanese patients with hypophosphatemic rickets. AB - Hypophosphatemic rickets is a genetic disorder commonly associated with renal phosphate wasting and bone deformities. The PHEX gene (phosphate regulating gene with homologies to endopeptidases on the X chromosome) encodes a 749-amino acid protein that putatively consists of an intracellular, transmembrane, and extracellular domain. PHEX mutations have been observed in 60-80% of hypophosphatemic rickets patients. In this study, we report 2 de novo novel mutations in 2 Taiwanese girls with clinical characteristics of hypophosphatemic rickets. The presenting phenotype of lower extremity deformities and short stature was suggestive of the diagnosis. Primers flanking 22 exons were used to amplify DNA by polymerase chain reaction. The results by direct DNA sequencing of case 1 revealed a C to T transition changing glutamine at codon 224 in exon 6 to a stop codon (Q224X). The result of case 2 showed a 2-base pair deletion (2090delGA) and resulted in a frameshift and premature termination of codon (PTC+19aa). Both mutations presumably result in a truncated protein, leading to loss of function of PHEX. This is the first report of PHEX gene mutation in the Taiwanese population. PMID- 15818437 TI - Fetus in fetu in an adult: diagnosis by computed tomography imaging. AB - The term "fetus-in-fetu" refers to a monozygotic, diamniotic twin that is incorporated into the body of another sibling early in embryonic development. A fetus in fetu is a differentiated mass that, in contrast to a teratoma, has a discrete spinal column and well developed internal organs. Most cases of fetus-in fetu are discovered as a retroperitoneal mass in the first year of life, and the condition is unusual in adulthood. We describe a case of fetus-in-fetu incidentally discovered in a 39-year-old man who complained of mild epigastralgia. Abdominal ultrasound revealed a mass at the right upper quadrant of the abdomen. Fetus-in-fetu was diagnosed by computed tomography as an incidental finding, which was not clearly related to his abdominal discomfort. The fetus-in-fetu was treated conservatively. His radiologic as well as clinical condition remained unchanged during 4 years of follow-up. PMID- 15818438 TI - Isolation and identification of Escherichia coli O157:H7 in a Taiwanese patient with bloody diarrhea and acute renal failure. AB - Escherichia coli O157:H7 is a frequent foodborne pathogen in many developed countries. Outbreaks of this infection have been reported in countries all over the world. The first clinical case of E. coli O157:H7 infection from Taiwan was reported in a 6- year-old boy who had returned from USA in August 2001. In this paper, we describe the results of the isolation and identification of this strain, and molecular typing for comparison with previously reported strains. Biochemical and molecular biological tests were used to confirm that this patient, who developed bloody diarrhea and kidney failure as a result of the infection, was indeed infected with E. coli O157:H7. None of the patients' close contacts were affected. Molecular typing by use of pulsed-field gel electrophoresis revealed this clinical strain to have a unique genotype, which is different from all available clinical strains reported from Japan and environmental strains reported from Taiwan. America Type Culture Collection reference strains and an out-break strain from USA had the nearest relationships with this clinical isolate. Molecular typing showed that this infection by E. coli O157:H7 was not derived from the local environmental strains and was acquired during overseas travel. PMID- 15818439 TI - Androgenetic alopecia and current methods of treatment. AB - Androgenetic alopecia (AGA) is a common dermatological condition affecting both men and women. In the case of men, up to 30% over the age of 30 and more than 50% over the age of 50 are affected. AGA also affects women although clinical signs are usually milder and associated with diffuse thinning of the scalp hair. AGA invariably causes serious psychological problems especially in women. By far the most promising approaches to the treatment of baldness in men are drug therapies, such as topical minoxidil and finasteride administered systemically. Mild to moderate AGA in women can be treated with antiandrogens and/or topical minoxidil with good results in many cases. PMID- 15818440 TI - Dermatological aspects of cerebrovascular diseases. AB - Neurological symptoms are sometimes triggered by the same mechanisms as are skin manifestations. They include genetic conditions like the epidermal nevus syndrome, the Sneddon syndrome, Fabry disease and others, as well as certain inflammatory disorders like erythematous lupus, Bechet disease. Basically all conditions giving rise to anticoagulation processes may cause simultaneously neurological and cutaneous manifestations. Cerebrovascular stroke is the third most common condition of death in the developed world after cancer and ischemic heart disease. The mechanisms responsible for development of skin manifestations in patients afflicted by stroke are shortly reviewed. Stroke may also influence the already existent skin diseases. PMID- 15818441 TI - Fabry disease. A case report. AB - Fabry disease is an under-recognized X-linked recessive lysosomal storage disorder resulting from the deficient activity of the enzyme alpha-galactosidase A (alpha-Gal A). The first case of Fabry disease in Slovenia was diagnosed in 1991. This 46 year-old male was referred for dermatologic evaluation of a purpura on his abdomen. He was being treated for proteinuria and cardiac symptoms. The diagnosis of angiokeratoma corporis diffusa (Fabry disease) was made clinically and confirmed by demonstration of the deficient leukocyte alpha-Gal A activity. The patient subsequently developed cerebrovascular symptoms, coronary disease, and renal failure, and died from a recurrent myocardial infarction. Family studies identified several other affected males and carrier female relatives with this X-linked recessive disorder. This case illustrates the typical multi manifestations of this inherited disease which now can be safely and effectively treated by enzyme replacement therapy. Early diagnosis is important for the most effective treatment of this disease. PMID- 15818442 TI - Angiosarcoma arising in sclerodermatous skin. AB - We report a case of cutaneous angiosarcoma in a 77-year-old female patient with systemic sclerosis. The tumor developed around a large telangiectasia in the left temporal region. Later on, extensive asymptomatic redness and edema with several nodules over the whole left side of the face developed. Since the conservative therapy failed to produce any improvement, a diagnostic skin biopsy was performed. The pathohistological diagnosis was moderately differentiated angiosarcoma. The patient was treated with a combination of chemotherapy and electron beam irradiation. Despite a notable cutaneous improvement, the control check-up revealed the presence of metastases of the lung and liver. This is the first report of cutaneous angiosarcoma occurring in sclerodermatous skin. The possible pathogenesis of this rare tumorogenic transformation of soft tissues is discussed. PMID- 15818443 TI - Peutz-Jeghers syndrome. A case report. AB - Peutz-Jeghers syndrome (PJS) is an unusual hamartomatous polyposis of the gastro intestinal (GI) tract, with pigmentation around lips and macules on the buccal mucosa. The case of a 10-year-old girl who presented with intussusception is reported. A polyp was found to be the cause of an invagination. Histologically it was a hamartoma. PJS is a rare syndrome inherited in an autosomal dominant pattern. Most patients have recurrent episodes of polyp induced bowel intussusception which requires repeated laparotomies. In addition, these patients have an increased risk of malignant disease in gastrointestinal and also non gastrointestinal sites. To prevent cancer and short bowel syndrome, aggressive screening is recommended. Upper and lower endoscopy should be performed every two years from 10 years of age. Extra-intestinal surveillance for cancers, including abdominal and pelvic ultrasound, as well as testicular and breast examinations once yearly should be introduced in the second decade of life. PMID- 15818444 TI - Idiopathic eruptive macular pigmentation. AB - We present the case of a 10-year-old girl with a six months history of disseminated asymptomatic, brown pigmented macules on the trunk and proximal parts of the extremities. The clinical picture, histological findings, and the course of disease were similar to those of idiopathic eruptive macular pigmentation. The cutaneous lesions gradually disappeared over the next two years without any treatment, and no relapse occurred. The knowledge of this disease is important in order to avoid unnecessary treatment as spontaneous resolution of the lesions may be expected within months or a few years. The spontaneous regression without any treatment is an additional diagnostic criterion. PMID- 15818445 TI - Association of variants in angiotensin-converting enzyme and endothelin-1 genes with phototherapy in cutaneous T-cell lymphoma. AB - BACKGROUND: The cutaneous T-cell lymphoma (CTCL) is a disease characterised by cutaneous infiltrates of malignant, clonally expanded T-cells. Because individual genetic determination of angiogenetic and antioxidant properties of blood vessels could be partly responsible for phototherapy in CTCL patients, three polymorphisms in angiotensin converting enzyme and endothelin-1 genes were determined. METHODS: 77 patients with CTCL, diagnosed and treated at the First Dermatological Clinic of St. Ann's Faculty Hospital Brno (46 men and 31 women, median age 62, range 26-80 years) were compared to a control non-CTCL group of the similar age and gender distribution (n=203: 137 men and 66 women, median age 54, range 27-86 years) with negative family history of severe skin diseases and without signs of malignancy. Diagnosis of CTCL was verified according to the clinical picture and histologically. The genotype distributions and allelic frequencies between CTCL with phototherapy and those without phototherapy were compared. RESULTS: Significant differences were found in genotype distributions of I/D ACE polymorphism between CTCL patients treated with phototherapy and those treated without it. Heterozygote ID was more frequent in the group treated with phototherapy (25/13 vs. 12/27, OR=4.33, 95% confidential interval 1.67-11.24, P=0.02. The 4A4A variant of -3A/-4A EDN1 is more frequent in patients treated with phototherapy (8/30 vs. 1/38, OR=10.13, 95% confidential interval 1.20-85.55, P=0.01). The GA and AA genotypes of G8002A EDN1 polymorphism are more frequent in CTCL patients treated with phototherapy compared to those without it (15/23 vs. 7/32, OR=2.98, 95% confidential interval 1.05-8.48, P=0.03). DISCUSSION: Some polymorphic variants in ACE and EDN1 genes (a heterozygote ID in I/D ACE, a homozygote -4A-4A in -3A /-4A EDN1 and genotypes GA and GG in G8002A EDN1) seem to carry an advantage for phototherapy effectiveness in patients with CTCL. PMID- 15818446 TI - Successful treatment of chronic venous leg ulcers with lyophilized cultured epidermal allografts. AB - The case of a 67-year-old woman with chronic venous leg ulcers and severe gonarthrosis is described. In spite of intensive therapy, the leg ulcers had persisted for 4 years and made the intended orthopaedic operation of the right knee impossible. The patient was treated with lyophilized cultured epidermal allografts and her leg ulcers healed within 40 days. Lyophilized cultured epidermal allografts represent a modern type of active wound dressing that leads to rapid healing of chronic venous leg ulcers and enables patients to undergo surgical intervention. PMID- 15818447 TI - The multifunctional role of leukaemia inhibitory factor in cutaneous biology. AB - Leukaemia Inhibitory Factor (LIF) is a polyfunctional cytokine, that belongs to the family of haemopoietic growth factors. LIF plays a role in growth-promotion and differentiation, regulates calcium and bone metabolism, induces acute phase proteins and causes cachexia in organisms with neoplastic disorders. LIF is also to be found in normal skin, where it may be involved in the differentiation process of keratinocytes. In addition, recent data in medical literature indicates that LIF is engaged in the pathogenesis of some skin disorders as well. It has been clearly demonstrated that LIF may act as a proinflammatory cytokine. In allergic contact dermatitis, the expression of LIF mRNA is augmented to a significant degree, indicating that LIF may play a role in the early phase of allergic contact dermatitis. LIF also plays an important role in psoriatic lesions. As the mechanism is not yet fully understood, however, it is hypothesized that the LIF function in psoriatic processes is solely connected with IL-8, as it is known that LIF is able to induce the release of IL-8. Also, some reports have suggested that LIF may also play a role in the carcinogenesis of the skin. PMID- 15818448 TI - Lipoid proteinosis. AB - The case of a 37-year-old woman with lipoid proteinosis is presented. The first symptoms of hoarseness of voice occurred in puberty. Additional symptoms included beaded eyelid papules, macroglossia with yellowish papules, yellowish deposits of soft palate, popular eruption in the axillary area and tuberous nodules on the elbows. PMID- 15818449 TI - Photo-onycholysis: two cases induced by doxycycline. AB - Photo-onycholysis is a phototoxic reaction, which is usually drug-induced. It consists of the separation of the nail from the nail bed due to ultraviolet radiation. We report two cases of female patients who developed distal onycholysis while receiving doxycycline. Among the drugs that cause photo onycholysis, the most frequently cited are tetracyclines, psoralens and fluoroquinolones. Photo-onycholysis is often distal, half-moon shaped and can be surrounded by pigmentation. Spontaneous recovery follows within a few months of discontinuing the drug. PMID- 15818450 TI - Statins: novel weapons against granulomatous disorders and HIV infection? PMID- 15818451 TI - Proposal for an annual international day for the best journals of dermatology, and the best dermatological articles. PMID- 15818452 TI - Safety of contrast venography prior to caval interruption in patients with renal insufficiency. AB - We undertook this study to determine whether the use of contrast venography would adversely affect renal function in patients with renal insufficiency requiring caval interruption. We conducted a retrospective review of all inferior vena cava (IVC) filters inserted at our institution over a 2-year period (January 2002 to January 2004). The indication for caval interruption, insertion technique, type of filter used, pre- and postintervention creatinine level, and the presence of diabetes and hypertension were analyzed. A total of 282 IVC filters were inserted, with 38 of them placed in patients with renal insufficiency as defined by a serum creatinine level of > 1.5 mg/dL. Contrast venography with 15 to 30 mL of iohexol (Omnipaque 300) was used in all cases, and no special measures other than proper hydration were used for renal protection. All filters were successfully deployed. The mean +/- SD preintervention creatinine level was 2.38 +/- 0.79 mg/dL. The mean +/- SD postintervention creatinine levels at 2 and 30 days were 2.26 +/- 0.45 mg/dL and 2.12 +/- 0.94 mg/dL, respectively. No patients required hemodialysis following caval interruption, and no adverse effect on renal function was noted. Contrast venography accurately delineates venous anatomy and facilitates proper caval filter placement with no apparent adverse effect on renal function. We believe contrast venography is safe even in the presence of renal insufficiency. PMID- 15818453 TI - Diameter of the infrarenal abdominal aorta in children with cerebral palsy. AB - The diameters of the infrarenal abdominal aorta by ultrasonography have not yet been reported in children with cerebral palsy (CP). Because the demand for the surgical applications of the abdominal aorta probably increases in the CP population owing to their prolonged life-span, we planned a study to assess the diameter of the infrarenal abdominal aorta by ultrasonography in children with CP and to compare them with those of healthy subjects. The study comprised 60 children with CP and 95 age- and sex-matched healthy children. The diameter of the infrarenal abdominal aorta was measured by B-mode ultrasonography. The correlation between the aortic diameter, weight, height, body surface area (BSA), and body mass index was studied. The diameter of the infrarenal abdominal aorta was found to be smaller in children with CP than in healthy children (p < 0.05). There was a significant correlation between the aortic diameter, weight, height, and BSA in both groups (p < 0.05). When a vascular surgical treatment is needed in children with CP, the probability of a small-diameter infrarenal abdominal aorta should be considered. PMID- 15818454 TI - Management of infected grafts and aneurysms of the aorta. AB - Seventeen patients treated for infected grafts (11/17) or aneurysms (6/17) of the aorta between 1998 and 2003 were reviewed to evaluate our experience with aortic infection. The causative organisms were identified in 12 patients (71%), with 5 (29%) having methicillin-resistant Staphylococcus aureus. A periaortic abscess occurred in eight patients, and all of them were associated with infected grafts. Surgical treatment included cryopreserved allograft replacement in eight patients, prosthetic graft replacement in four patients, and drainage with or without omental wrapping in five patients. One patient was still hospitalized at the end of the study period. Five patients with infected grafts died after the operation during the initial hospitalization. No early mortality occurred in the aneurysm group. The early mortality rate was 31% for all patients, 50% for the graft group, and 63% for patients with a periaortie abscess. Another patient with an infected aneurysm died of arrhythmia after discharge from the initial hospitalization, Ten patients are still alive without evidence of reinfection. The early mortality rate for patients with infected aortic grafts is higher than that for those with infected aneurysms, especially when a periaortic abscess accompanies them. However, the late outcome is favorable, with no reinfection or late treatment-related deaths. PMID- 15818455 TI - Prognostic factors for mesenteric infarction: multivariate analysis of 187 patients with regard to patient age. AB - Between 1980 and 2002 a total of 187 patients (105 women, 82 men) with a mean age of 71.5 years underwent surgery for acute mesenteric infarction (AMI) in our department. Arterial occlusion was found in 140 patients, splanchnic vein thrombosis in 22, and nonocclusive AMI in 25. We performed bowel resection in 61 patients, vascular surgery in 32, a combination of the two in 15, and an exploratory laparotomy in 79. The mortality rate was 70.6% (73.9% in patients >70 years and 65.2% in those <70). AMI is still a disease with a consistently high mortality rate (60-80%). Multivariate prognostic factors include the time until surgery and a markedly elevated white blood cell count and serum lactate level. Mortality and secondary disease rates are higher in the elderly, and the fact that they have to wait much longer for an operation than younger patients may be a factor. If surgery is performed early, the survival rate increases regardless of age. The most important prognostic factor and the only one that can be influenced by the surgeon remains the time interval between the onset of symptoms and surgery, Therefore diagnostic and therapeutic intervention should be performed as early as possible in patients with suspected AMI. PMID- 15818456 TI - Vascular closure stapler clips versus polypropylene sutures in end-to-end anastomoses of growing arteries and veins. AB - Alternatives are still being sought in vascular surgery to avoid the complications caused in growing vessels by the various suture techniques and materials currently in use. A major complication is arrested growth following anastomosis. Opinions differ widely regarding the best approach to this problem, and research is currently underway at our center on the latest generation of clip appliers (vascular closure stapler, or VCS) and their potential application as an alternative to the conventional suture technique in pediatric vascular surgery. This study compared vascular growth and reendothelialization following anastomosis of growing arteries and veins using VCS clips versus the conventional interrupted polypropylene suture technique. Thirty 55-day-old lambs underwent end to-end anastomosis of the carotid artery and jugular vein. Serial ultrasonography and angiography were carried out over the ensuing 6-month growth period, after which the lambs were euthanized. Gross examination of the anastomosis site was followed by histologic examination involving light microscopy, immunohistochemical techniques, and transmission electron microscopy. Although both VCS clips and polypropylene sutures allowed transverse vessel growth, the clips did not penetrate the intima and thus provided enhanced reendothelialization. The satisfactory results obtained here, as in earlier studies, increasingly point to VCS clips as the method of choice in pediatric vascular surgery. PMID- 15818457 TI - Overexpression of MMP9 and tissue factor in unstable carotid plaques associated with Chlamydia pneumoniae, inflammation, and apoptosis. AB - Tissue remodeling by matrix metalloproteinases (MMPs) and plasminogen activators such as tissue factor (TF) is postulated to be involved in the pathogenesis of atherosclerosis. The in situ expression of MMP9 and TF in unstable atherosclerotic plaques has not been examined in detail. Moreover, interference of tissue remodeling by vascular inflammation, apoptosis, and Chlamydia pneumoniae inside plaque subregions is unclear. A total of 40 autopsy carotid arteries (controls) and 20 atherosclerotic carotid endarterectomy specimens (with type VI lesions, according to the American Heart Association classification) from stroke patients were analyzed for expression of MMP9 and TF using in situ techniques. The data on tissue remodeling were correlated with the presence of inflammatory cells (T cells, B-cells, macrophages), apoptosis, and the presence of C. pneumoniae using immunohistochemistry and Western blot analyses. We found a significant overexpression of MMP9 and TF in progressive atherosclerotic carotid arteries, especially in the shoulder and cap subregions (both p < 0.05). Expression of MMP9 and TF correlated significantly with T-cell and macrophage infiltrates as well as with apoptosis (p < 0.05). C. pneumoniae infection was significantly associated with elevated TF expression (p < 0.01) but not with MMP9. MMP9 and TF are thus significantly overexpressed in progressive atherosclerotic plaques, and their relevant subregions (shoulder and cap) are involved in plaque instability. This process is associated with local inflammatory cell infiltrates and apoptosis, which might be influenced by infectious agents such as C. pneumoniae. PMID- 15818458 TI - Aneurysm sac thrombus load predicts type II endoleaks after endovascular aneurysm repair. AB - Type II endoleaks are associated with the absence of aneurysm shrinkage after endovascular abdominal aortic aneurysm repair (EVAR). This study aims at determining the predictability of this complication, whose potential risk factors have been the subject of conflicting reports. Preoperative computed tomography (CT) scans of 178 patients who underwent EVAR for true infrarenal abdominal aortic aneurysms between January 20, and April 17, 2003, with a minimum follow-up of 30 days, were reviewed. The following information was retrieved: maximum aneurysm diameter, aneurysm thrombus load (maximum thickness, percentage of sac circumference wall coverage, percentage of maximum sac area occupancy); number, diameter, and nature (lumbar, inferior mesenteric, accessory renal, middle sacral) of patent aortic side-branch arteries; thrombus thickness at each aortic branch ostium, and aneurysm diameter at that level. Postoperative CT and duplex scans supplemented with angiography in selected cases were reviewed for the presence of a type II endoleak observable beyond the 30th postoperative day. Logistic regression was used to assess the association of each variable with this outcome. There were 38 (21.3%) patients with type II endoleaks after the 30th postoperative day. The median follow-up was 12 months (range 1-65 months). By univariate analysis, the following variables significantly decreased the risk of a type II endoleak: thrombus maximum thickness [odds ratio (OR) 0.77 for a 5 mm increase, p = 0.009], mean thrombus thickness at aortic side-branches ostia (OR 0.65 for a 1 mm increase, p = 0.0006), thrombus-occupied percentage of maximum aneurysm area (OR 0.72 for a 10% increase, p < 0.0001), percentage of thrombus lined aneurysm wall (OR 0.53 for a 25% increase, p < 0.0001). The presence of a patent inferior mesenteric artery (OR 6.84, p < 0.01) and the number of patent aortic side-branches (OR 1.37 for each additional vessel, p = 0.002) significantly increased the risk of detecting a late type II endoleak. Aneurysm and aortic side-branch diameters did not have any impact. In a multiple logistic regression model (whole model p < 0.0001), the thrombus-occupied percentage of maximum aneurysm area (OR 0.74 for a 10% increase, p < 0.0005) and the number of patent aortic side-branches (OR 1.31 for each additional vessel, p = 0.009) remained independent predictors of type II endoleaks. The simple measure of the proportion of maximum aneurysm area occupied by thrombus may be a useful way to identify patients at high risk of a persistent type II endoleak. Patients with low preoperative sac thrombus load should be followed with a high degree of suspicion for this complication. PMID- 15818459 TI - Effects of cilostazol on human venous smooth muscle. AB - In this study, we evaluated the effect of therapeutic doses of cilostazol on human venous smooth muscle. Saphenous vein rings (two to four per patient sample) were suspended in tissue baths for isometric tension recordings. At the beginning of the experiment, optimal tension for isometric contraction was achieved for each ring in a stepwise fashion in the presence of norepinephrine (10(-2) M). Norepinepherine was then added cumulatively in half-molar increments and isometric tension developed by the rings was measured, thereby obtaining a dose response curve. Following washout and reequilibration, the rings were precontracted with a 30-50% submaximal dose of norepinepherine determined from the dose-response curve and allowed to contract until a stable plateau was reached. Cilostazol was then added in a cumulative manner (680-2,720 microg/L), and the tension generated was recorded. A total of 76 venous rings were tested, and all relaxed in the presence of cilostazol. The amount of relaxation increased as the concentration of cilostazol increased. Relaxation of 15 +/- 1.9% (mean +/- SEM) at low cilostazol doses (680 microg/L) to 37+/-3% at high cilostazol doses (2,720 microg/L) was demonstrated. A second finding of this study was demonstrated when the patient samples were divided according to the presence or absence of risk factors for arteriosclerosis. The specific risk factors examined included diabetes mellitus, smoking, hypercholesterolemia, and hypertension. The presence or absence of hypertension (n = 52) or hypercholesterolemia (n = 18) did not affect the amount of relaxation of the venous rings. Smokers (n = 46) had less relaxation 16 +/- 2.4% (680 microg/L) to 41 +/- 3.6% (2,720 microg/L) compared to nonsmokers (n = 53) who relaxed 22 +/- 3.5% (680 microg/L) to 48 +/- 5.7% (2720 microg/L). This did not reach statistical significance at any concentration cilostazol (p = 0.11-0.18). Diabetics (n = 53) did have statistically significantly less relaxation at every concentration of cilostazol compared to nondiabetics (n = 11, p < 0.05). All venous rings relaxed in the presence of cilostazol. Veins of nondiabetics relaxed statistically significantly more than those of diabetics. Smokers had less relaxation than non-smokers, but this was not statistically significant. We are the first to demonstrate that human venous smooth muscle cells undergo relaxation when exposed to therapeutic concentrations of cilostazol. PMID- 15818460 TI - Nitric oxide mediates the effect of fluvastatin on intercellular adhesion molecule-1 and platelet endothelial cell adhesion molecule-1 expression on human endothelial cells. AB - Leukocyte and platelet adhesion to endothelial cells, an early step in the pathogenesis of atherosclerosis, is mediated through adhesion molecules. It has been shown that statins decrease adhesion molecule expression. We examined the hypothesis that fluvastatin decreased intercellular adhesion molecule-1 (ICAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) expression through a nitric oxide-mediated pathway. Human iliac artery endothelial cells were exposed to fluvastatin in the presence or absence of 2 mM N-monomethyl-L-arginine (L NMMA). Flow cytometry analysis was used to measure ICAM-1 and PECAM-1 expression. In a separate experiment, confluent cell cultures were exposed in a serum-free medium to fluvastatin 20 microM, and the supernatant was collected for nitrate/nitrite determination after 6 and 48 hr of incubation. Protein was isolated and processed for immunoblotting with monoclonal antibodies specific for endothelial nitric oxide synthase (eNOS), Ser(1177)-phosphorylated eNOS, and AMP kinase. Relative band intensity was assessed with densitometry. Results are presented as the mean +/- standard deviation (SD), and p < 0.05 was considered significant. ICAM-1 and PECAM-1 were expressed constitutively. Human iliac artery endothelial cells (HIAECS) treated with 5 microM fluvastatin did not exhibit reduced expression of PECAM-1 or ICAM-1. Incubation with 10 microM fluvastatin reduced basal expression of both ICAM-1 and PECAM-1. Fluorescence intensity (FI) for these substance was as follows: 3638 +/- 1671, p = 0.01 and PECAM-1 vs. control FI 276 +/- 52 vs. 522 +/- 78, p = 0.02. In the presence of 2 mM L-NMMA, fluvastatin failed to decrease the expression of ICAM-1 (fluvastatin 10 microM + L-NMMA: FI was 3042 +/- 1378 vs. 3638 +/- 1671 for the control p = 0.01) or PECAM 1 (fluvastatin 10 microM + L-NMMA: FI was 415 +/- 188 vs. 522 +/- 78 for the control, p = 0.1). Incubation with 20 microM fluvastatin similarly reduced ICAM-1 expression (FI was 2014 +/- 1595 vs. 3638 +/- 1671 for the control, p = 0.02) and PECAM-1 expression (FI was 196 +/- 109 vs. 522 +/- 78 for the control, p = 0.02). This reduction was prevented in the presence of 2 mM L-NMMA. L-NMMA in a concentration of 2 mM had no significant effect on adhesion molecule expression (p > 0.05 for all comparisons of the control FI versus 2 mM L-NMMA mean FI). After a 48 hr incubation with 20 microM fluvastatin there was a 219 +/- 35% increase in the cell eNOS protein content (p = 0.01) and a 170 +/- 26% increase in the cell AMPK protein content (p = 0.02). Ser(1177)-phosphorylated eNOS protein levels were increased by 41 +/- 8% (p = 0.03). The nitric oxide concentration in the medium of the HIAEC treated with 20 microM fluvastatin for 48 hr was significantly higher than that in the control (p = 0.0004), pointing to increased production during the incubation period. Fluvastatin thus decreases basal expression of ICAM-1 and PECAM-1. Competitive inhibition of eNOS with L NMMA abolishes the effect of fluvastatin on ICAM-1 and PECAM-1 expression. The statin up-regulates eNOS and AMP kinase, one of the enzymes that activates eNOS via phosphorylation at Ser(1177). We have shown that after a 48-hr exposure to fluvastatin there is an increased amount of the phosphorylated enzyme in the endothelial cells. PMID- 15818461 TI - Percutaneous transluminal revascularization for iliac occlusive disease: long term outcomes in TransAtlantic Inter-Society Consensus A and B lesions. AB - Percutaneous transluminal intervention for atherosclerotic iliac occlusive disease is now commonplace. We examine the long-term outcomes of TransAtlantic Inter-Society Consensus (TASC) A and B lesions. We performed a retrospective anonymous analysis of records from patients who underwent iliac artery angioplasty with or without stenting between January 1990 and June 1999. Indications for intervention were symptomatic claudication (77%) or critical ischemia (23%). Altogether, 276 patients (all men; average age 64 +/- 11 years range 32-87 years) underwent 394 interventions. Co-morbidities included hypertension (61%), hypercholesterolemia (45%), diabetes (28%), and chronic renal insufficiency (26%). A total of the 62% of the lesions were TASC category A, and the remainder were category B. Of the 394 primary interventions, 51% included placement of stents. Technical success (defined by < 30% residual stenosis) was achieved in 98% of treated vessels. The procedure-related mortality rate was 1.8% at 30 days and 4.7% at 90 days; the procedure-related complication rate was 7%. Hemodynamic success (defined as a rise in the ankle/branchial index > 0.15) was achieved in 82%. The average Society for Vascular Surgery symptom score was 3.4 +/- 0.9 before intervention, which improved to 1.9 +/- 0.8 following intervention. Within 3 months, 84% of patients demonstrated clinical improvement. Patient survival by life-table analysis was 38% at 10 years. The cumulative assisted patency rate was 71 +/- 7% at 10 years. The presence of two-vessel femoral runoff, two or more patent tibial vessels, or both was associated with improved patency. Limb salvage was 95 +/- 2% and 87 +/- 9% at 5 and 10 years, respectively. Using Cox proportional hazards analysis, the presence of hypertension, hypercholesterolemia, or chronic renal insufficiency was associated with the occurrence of primary failure, whereas increased patency intervals were associated with the presence of immediate hemodynamic improvement. Use of a stent did not influence outcome. Endoluminal iliac intervention for TASC A and B lesions is a safe, durable intervention in patients with good femoral and tibial runoff. The presence of hypertension, hypercholesterolemia, or poor tibial runoff is associated with failure. PMID- 15818462 TI - Surgical treatment of inflammatory abdominal aortic aneurysms: a long-term follow up of 19 patients. AB - The prevalence of inflammatory abdominal aortic aneurysms (IAAA) in autopsy material ranges between 2.5 and 10% of all aneurysms. Clinical findings, the distinction between inflammatory and degenerative aneurysms, and epidemiological data are uncertain, and only a few long-term follow-up studies of patients after surgical treatment of IAAAs exist. In this study, 19 patients underwent either emergency or elective surgery for IAAA during the 10-year period between 1983 and 1993 at Helsinki University Central Hospital. Demographics, symptoms, and operative and follow-up data were collected retrospectively with emphasis on the long-term outcome of IAAA. Causes of late death were available from hospital records and the central statistical office of Finland. For survival analysis we compared ruptured versus nonruptured and emergency versus elective cases of IAAAs. Mean follow-up for the 18 surviving patients (1 hospital death) was 7.4 years. One patient (5%) died of a long-term complication of the aneurysmal disease. There was no statistically significant difference in survival rates for emergency versus elective surgery cases or ruptured versus nonruptured aneurysms. The most common cause of late death was myocardial infarction. The hospital stay mortality (5%) and morbidity (31%), and the survival rate of 26% at 5 years for ruptured and 65% and 43% for nonruptured IAAAs at 5 years and 10 years, respectively, are comparable to normal AAA survival rates. These findings show that surgery is recommended, especially as ruptures also occur in this subgroup of aneurysms. PMID- 15818463 TI - Should SAGES take the lead in a little known resident training challenge? PMID- 15818464 TI - Very close relationship of the chloroplast genomes among Saccharum species. AB - We recently determined the complete sequence of the sugarcane chloroplast genome. Here, we have used the information for a comprehensive phylogenetic analysis of the genus Saccharum, using all six species (13 accessions). The polymorphisms between sugarcane and maize in 26 chloroplast genome regions were used for the analysis. In 18 of the 26 regions (a total of 5,381 bp), we found 41 mutations involving 17 substitutions, three inversions, six insertion/deletion mutations, and 15 simple sequence repeat length polymorphisms. Based on these results, we calculated a phylogenetic tree of the genus Saccharum, in which all six species are clearly separated. By the analysis, (1) S. sinense and S. barberi, which have identical sequences, belong to the same clade, whereas the other four species, S. officinarum, S. robustum, S. edule, and S. spontaneum, form an independent clade; (2) S. spontaneum has a paraphyletic relationship with the other five species; and (3) no or very low intraspecific variation was observed in S. officinarum, S. robustum, S. sinense, S. barberi, and S. edule, whereas higher intraspecific variation was observed in S. spontaneum. Based on the number of nucleotide substitutions, the divergence time between S. officinarum and S. spontaneum, and between S. officinarum and maize were calculated to be about 730-780 thousand years ago and about 5.9 million years ago, respectively. These results suggest that the cytoplasm of Saccharum species are very closely related. PMID- 15818465 TI - Prospective, randomized, double-blind study of safety and tolerability of intravesical resiniferatoxin (RTX) in interstitial cystitis (IC). AB - OBJECTIVE: To determine the safety and tolerability of intravesical resiniferatoxin (RTX) in interstitial cystitis (IC) patients. MATERIALS AND METHODS: IC patients were instilled with 50 cc of test solution containing either placebo, 0.05 microM or 0.10 microM RTX in the bladder. Plasma concentration of RTX and its degradant resiniferonol 9-, 13-, 14-orthophenylacetate was measured. Immediate post-treatment blood sampling and cystoscopy were performed. Symptoms were evaluated before treatment, at 4- and at 12-week follow-ups, using VAS indicator for pain, voiding diary, and O'Leary's IC symptom/problem indices. RESULTS: Among 22 patients observed (ten in 0.10 microM RTX, eight in 0.05 microM RTX, and four in placebo groups), the most commonly reported adverse event was pain during instillation (80.0%, 87.5%, and 25.0%). No serious adverse events were reported. CONCLUSIONS: Use of intravesical RTX in IC patients is associated with important tolerability issues but safe at 0.10 microM and 0.05 microM. PMID- 15818466 TI - Plexins: axon guidance and signal transduction. AB - Axon guidance represents a key stage in the formation of neuronal network. Axons are guided by a variety of guidance factors, such as semaphorins, ephrins and netrin. Plexins function as receptors for the repulsive axonal guidance molecules semaphorins. Intracellular domains of plexins are responsible for initiating cellular signal transduction inducing axon repulsion. Recent advances have revealed molecular mechanisms for plexin-mediated cytoskeletal reorganization, leading to repulsive responses, and small GTPases play important roles in this signaling. Plexin-B1 activates Rho through Rho-specific guanine nucleotide exchange factors, leading to neurite retraction. Plexin-B1 possesses an intrinsic GTPase-activating protein activity for R-Ras and induces growth cone collapse through R-Ras inactivation. In this review we survey current understanding of the signaling mechanisms of plexins. PMID- 15818467 TI - Characterization of the rdar morphotype, a multicellular behaviour in Enterobacteriaceae. AB - The rdar morphotype, a multicellular behaviour of Salmonella enterica and Escherichia coli is characterized by the expression of the adhesive extracellular matrix components cellulose and curli fimbriae. The response regulator CsgD, which transcriptionally activates the biosynthesis of the exopolysaccharide cellulose and curli, also transforms cell physiology to the multicellular state. However, the only role of CsgD in cellulose biosynthesis is the activation of AdrA, a GGDEF domain protein that mediates production of the allosteric activator cyclic-di-(3'-5')guanylic acid (c-di-GMP). In S. enterica serovar Typhimurium a regulatory network consisting of 19 GGDEF/EAL domain-containing proteins tightly controls the concentration of c-di-GMP. c-di-GMP not only regulates the expression of cellulose, but also stimulates expression of adhesive curli and represses various modes of motility. Functions of characterized GGDEF and EAL domain proteins, as well as database searches, point to a global role for c-di GMP as a novel secondary messenger that regulates a variety of cellular functions in response to diverse environmental stimuli already in the deepest roots of the prokaryotes. PMID- 15818468 TI - Decision-making at the surface of the intact or barrier disrupted skin: potential applications for vaccination or therapy. AB - The skin is a highly accessible organ and constitutes an active immunological site. Both these properties make this surface an attractive route for what promises to be a cost-effective, simple, practical and needle-free delivery of vaccines and immunomodulators. Less obvious is the fact that the state of the skin barrier can influence quantitative and qualitative aspects of antigen specific immune responses. The everyday decision-making at the skin epithelium concerns the choice between the induction of an immune response and the establishment of a state of non-responsiveness (tolerance). This decision is influenced by various factors such as the dose, the route (intact vs barrier disrupted skin), the cytokine microenvironment and the nature of the antigenic stimulus. By increasing our understanding of how immune responses are regulated in the epidermis we can envisage the development of immunisation protocols aimed at eliciting a protective immune response or inducing tolerance, with direct applications to preventive or therapeutic vaccination, respectively. PMID- 15818469 TI - Familial hypobetalipoproteinemia: genetics and metabolism. AB - Familial hypobetalipoproteinemia (FHBL), an autosomal dominant disorder, is defined as <5th percentile LDL-cholesterol or apolipoprotein (apo) B in the plasma. FHBL subjects are generally heterozygous and asymptomatic. Three genetic forms exist: (i) premature stop codon specifying mutations of APOB; (ii) FHBL linked to a susceptibility locus on the chromosome 3p21; and (iii) FHBL linked neither to APOB nor to the chromosome 3p21. In heterozygous apoB-defective FHBL, the hepatic VLDL export system is defective because apoB 100, the product of the normal allele, is produced at approximately 25% of normal rate, and truncated apoB is cleared too rapidly. The reduced capacity for hepatic triglyceride export increases hepatic fat three-fold. Indexes of adiposity and insulin action are similar to controls. 'Knock-in' mouse models of apoB truncations resemble human FHBL phenotypes. Liver fat in the chromosome 3p21-linked FHBL is normal. Elucidation of the genetic basis of the non-apoB FHBL could uncover attractive targets for lipid-lowering therapy. (See note added in proof.). PMID- 15818470 TI - Paraphyly of Cyrtomium (Dryopteridaceae): evidence from rbcL and trnL-F sequence data. AB - Cyrtomium is an Asiatic genus characterized by anastomosing veins with included veinlets, and comprises about 40 species. We sequenced rbcL and trnL-F sequences of 19 species of Cyrtomium and eight species from related genera in order to elucidate a molecular phylogeny of the genus using maximum-parsimony methods. The phylogenetic trees did not agree with traditional classifications. Cyrtomium was resolved as paraphyletic, and a clade including subseries Balansana of Cyrtomium, Cyrtogonellum, Polystichum subacutidens and Cyrtomidictyum (the BCPC clade) and a second one containing Cyrtomium sensu stricto were monophyletic. The results also implied that: (1) C. uniseriale was synonymous with C. balansae; (2) C. falcatum was likely the female parent of C. devexiscapulae; and (3) based on the rbcL and trnL-F sequence data, C. nephrolepioides and C. grossum were the female parents of C. shingianum and C. chingianum, respectively, although other evidence is needed for the confirmation of this hypothesis. PMID- 15818471 TI - A Festschrift honoring Norman M. Rich. AB - A Festschrift honoring forty years of government services, twenty-five years as Founding Chairman of the Department of Surgery at the F. Edward Hebert School of Medicine of the Uniformed Services University of the Health Sciences, Bethesda, Maryland and the seventieth birthday of Norman M. Rich, MD, Facs, DMCC, was held on Friday 26 March, 2004 in conjunction with the 24th Annual USU Surgical Associates Day. This article describes that event and introduces the articles written in celebration of it. PMID- 15818472 TI - Virchow's Legacy: deep vein thrombosis and pulmonary embolism. PMID- 15818474 TI - Competition and coexistence of aerobic ammonium- and nitrite-oxidizing bacteria at low oxygen concentrations. AB - In natural and man-made ecosystems nitrifying bacteria experience frequent exposure to oxygen-limited conditions and thus have to compete for oxygen. In several reactor systems (retentostat, chemostat and sequencing batch reactors) it was possible to establish co-cultures of aerobic ammonium- and nitrite-oxidizing bacteria at very low oxygen concentrations (2-8 microM) provided that ammonium was the limiting N compound. When ammonia was in excess of oxygen, the nitrite oxidizing bacteria were washed out of the reactors, and ammonium was converted to mainly nitrite, nitric oxide and nitrous oxide by Nitrosomonas-related bacteria. The situation could be rapidly reversed by adjusting the oxygen to ammonium ratio in the reactor. In batch and continuous tests, no inhibitory effect of ammonium, nitric oxide or nitrous oxide on nitrite-oxidizing bacteria could be detected in our studies. The recently developed oxygen microsensors may be helpful to determine the kinetic parameters of the nitrifying bacteria, which are needed to make predictive kinetic models of their competition. PMID- 15818475 TI - Selection strategies for the establishment of recombinant Chinese hamster ovary cell line with dihydrofolate reductase-mediated gene amplification. AB - To evaluate the efficacy of selection strategies for recombinant Chinese hamster ovary (rCHO) clones undergone with dihydrofolate reductase-mediated gene amplification, rCHO cell lines producing a chimeric antibody were established using two strategies, one based on individual clones and the other based on cell pools. In a selection based on individual clones, cell cloning by limiting dilution method was performed twice, once after a round of selection of parental cell clones and once after obtaining high-producer clones. Thirty parental clones selected from 300 parental clones were cultivated independently throughout the gene amplification procedure. Using this labor-intensive strategy, it took approximately 17 weeks to obtain high-producing clones such as CS11-8 and CS18-3 clones. A selection based on cell pools, in which cell cloning was performed once at the final selection stage, required less effort and time to amplify large numbers of individual parental clones within the pool. However, high-producing clones were lost during the amplification procedure. The antibody expression level of high-producing clones such as PS7-2 and PS7-32 chosen on the basis of cell pools was less than one third of that of CS11-8 and CS18-3 clones. Taken together, a selection strategy based on individual clones is favored for establishment of high-producing rCHO clones because it is more efficient to perform cell cloning at the initial selection stage of parental cell clones. PMID- 15818476 TI - Quantification of an Eikelboom type 021N bulking event with fluorescence in situ hybridization and real-time PCR. AB - Primers targeting 16S rRNA genes were designed to detect and quantify Eikelboom type 021N organisms by real-time PCR. Eikelboom type 021N filamentous bulking was induced in a laboratory-scale sequencing batch reactor and the evolution of Eikelboom type 021N 16S rRNA and 16S rRNA genes was monitored. A significant correlation was found between the sludge volume index and the amount of these filamentous organisms present in the sludge (r (2)=94.6%, n=10, P<0.01), as measured by real-time PCR. The amount of Eikelboom type 021N 16S rRNA genes increased by a factor of 21 during the experiment, while the 16S rRNA increased by a factor of 33. Moreover, Eikelboom type 021N 16S rRNA increased with increased feeding frequency. It was observed that the RNA:DNA ratio peaked before the sludge volume index increased. In parallel, a fluorescence in situ hybridization study indicated a factor of four increase in the length of Eikelboom type 021N filaments, due to a factor of two increase in both length and number of Eikelboom type 021N filaments. Further, an increase in the fraction of filaments extending outside the activated sludge flocs was observed (19-55%). Monitoring of 16S rRNA genes and 16S rRNA of Eikelboom type 021N was shown to be valuable in evaluating activated sludge settling characteristics; and measuring RNA:DNA ratios may be used as an early warning tool for sludge bulking. PMID- 15818477 TI - Curdlan and other bacterial (1-->3)-beta-D-glucans. AB - Three structural classes of (1-->3)-beta-D-glucans are encountered in some important soil-dwelling, plant-associated or human pathogenic bacteria. Linear (1 ->3)-beta-glucans and side-chain-branched (1-->3,1-->2)-beta-glucans are major constituents of capsular materials, with roles in bacterial aggregation, virulence and carbohydrate storage. Cyclic (1-->3,1-->6)-beta-glucans are predominantly periplasmic, serving in osmotic adaptation. Curdlan, the linear (1- >3)-beta-glucan from Agrobacterium, has unique rheological and thermal gelling properties, with applications in the food industry and other sectors. This review includes information on the structure, properties and molecular genetics of the bacterial (1-->3)-beta-glucans, together with an overview of the physiology and biotechnology of curdlan production and applications of this biopolymer and its derivatives. PMID- 15818478 TI - Phase-inversion tissue harmonic imaging compared to fundamental B-mode ultrasound in the evaluation of the pathology of large and small bowel. AB - Our purpose was to compare phase-inversion harmonic imaging (PIHI) with conventional B-mode ultrasound (US) regarding image quality in the evaluation of bowel pathology. Forty-one patients prospectively underwent intestinal ultrasound scans with US and PIHI in randomly chosen order. Crucial technical parameters were standardized. Bowel morphology as well as perienteric pathology and complications were documented. In 24 cases, the ultrasound results were compared to those of other imaging modalities. Three radiologists evaluated (1) overall image quality, (2) lesion conspicuity and diagnostic confidence, and (3) detection of free fluid on hardcopy films. The ratings for image quality were compared using the two-sample paired t test for means and Bowker's test for symmetry (p=0.05). Compared to US, PIHI provided significantly better overall image quality, lesion conspicuity and diagnostic confidence, as well as better detection of free fluid (p< 0.05). Bowel wall pathology, detected by both modalities, showed good correlation to additional imaging modalities. In 12 patients (29.3%), a gain of crucial diagnostic information was observed with PIHI when compared to US. PIHI significantly enhances sonography of the intestine by offering better overall image quality, better visualization of bowel pathology and associated changes. Additionally, PIHI adds crucial diagnostic information in several patients. PMID- 15818479 TI - Optical quality of the ocular lens of the sea lamprey (Petromyzon marinus) during the mature and transformer periods of life. AB - While larval sea lampreys exist as eyeless filter feeders for several years, they transform into free-swimming juveniles (transformers) that attach parasitically to prey fish as they develop sexual maturity. This study examines lamprey lens development and optics and, since the lens is often the only refractive component of an aquatic eye, the data also provide an indication of visual ability during transformer and adult periods of life. Seven adult sea lampreys (0.40-0.55 m) and eight transformers (0.15-0.18 m) were sacrificed, the eyes removed and lenses dissected, measured, and placed in an automated laser scanning instrument. Back vertex focal length (spherical aberration) was measured for 14 beam positions across each lens by using a digital camera to record the position of the refracted beam. Transformer lenses exhibit positive spherical aberration, with average focal lengths varying from about 2.40 mm near the lens center and 1.06 mm at the lens periphery. On the other hand, the lenses from adults are largely corrected for spherical aberration, with average focal lengths varying from 2.19 mm to 2.44 mm. This result indicates that the younger lenses do not have a gradient refractive index necessary to mitigate the aberration and that further study of this model may reveal the relation between lens embryology and the development of such a gradient. PMID- 15818480 TI - Decrease of total activity with time at long distances from a nuclear accident or explosion. AB - Two data groups were analyzed: (1) the exposure rate in the former Czechoslovakia after the Chernobyl accident in 1986, and (2) the decrease of beta activity of an atmospheric fallout sample taken in Bratislava during 24 h on 30 May 1965. Both quantities decreased with the first power of time. This pattern of decrease is explained by applying the same mathematical formalism as is also used to describe the decrease in postnatal mortality with age. Following this formalism, the decrease of total activity with the first power of time could be seen as a consequence of a log-normal distribution of decay constants in the fallout. This differs slightly from earlier results that show the total activity decreasing with a power of 1.2 immediately after the nuclear explosion. PMID- 15818481 TI - Chromosomal aberrations in peripheral lymphocytes of patients treated with radium 224 for ankylosing spondylitis. AB - The aim of this study was to investigate the in vivo frequency of chromosomal aberrations (primarily dicentric chromosomes and chromatid breaks) potentially induced by (224)Ra alpha-radiation in peripheral lymphocytes. The study was designed to serve as a cytogenetic analysis along with the therapeutic procedure of ankylosing spondylitis patients who were undergoing a treatment with (224)Ra chloride. The total administered activity was 10 MBq, and the treatment followed a schedule of 10 i.v. injections per week, each with a dose of 1 MBq of (224)Ra. The calculation of absorbed doses delivered to the blood used the models suggested by the ICRP and yielded a value of 4.7 mGy/MBq. The frequency of chromosomal aberrations observed during the course of therapy was related to the blood dose. The frequency of dicentric chromosomes induced in vivo was found to agree well with the corresponding value of dicentrics induced in vitro. However- given that peripheral lymphocytes are in the cell cycle's G(0) stage--an unexpected increase with dose in the yield of chromatid breaks was observed, with about 95% of them occurring in cells without any other chromosome-type aberrations. Reasons for the production of chromatid breaks are discussed. PMID- 15818482 TI - Eggs and embryos in Xenoturbella (phylum uncertain) are not ingested prey. AB - Xenoturbella is an enigmatic animal that has puzzled science for almost a century. The eggs and embryos found in Xenoturbella have recently been interpreted as ingested prey. However, PCR on individual eggs as well as in situ hybridisation and in situ PCR unambiguously show that they are Xenoturbella's own. The eggs and embryos are individually enclosed within follicles with the same ultrastructure. The cortical granules in oocytes and eggs from Xenoturbella but not Nucula stained positively with an antiserum against Reissner's substance. The embryos incorporated 5-bromodeoxyuridine in vivo, i.e. they replicate their genome and are living. PMID- 15818483 TI - Divergent expression patterns of Sox9 duplicates in teleosts indicate a lineage specific subfunctionalization. AB - Sry-related HMG-box genes are key regulators of several developmental processes. Sox9 encodes a transcription factor required for cartilage formation and testis determination in mammals. In zebrafish (Danio rerio) and stickleback (Gasterosteus aculeatus) two co-orthologs of Sox9 are present. To date, only one Sox9 had been identified in medaka (Oryzias latipes). We have now isolated the second Sox9 gene. Sequence analysis, phylogenetic data, linkage mapping as well as expression pattern all together suggest that the medaka Sox9a and Sox9b are co orthologs. During embryogenesis, the expression pattern of Sox9a and Sox9b are distinct but overlap considerably in craniofacial cartilage elements. Comparing the zebrafish Sox9a and Sox9b expression patterns with medaka Sox9a and Sox9b expression domains revealed that some are identical but others are clearly different. We conclude that Sox9 regulatory subfunctions were not partitioned before divergence of the teleosts and evolved to lineage-specific expression domains. PMID- 15818484 TI - Analysis of the transcriptional activation domain of the Drosophila tango bHLH PAS transcription factor. AB - Basic-helix-loop-helix-PAS transcription factors play important roles in diverse biological processes including cellular differentiation and specification, oxygen tension regulation and dioxin metabolism. Drosophila tango is orthologous to mammalian Arnt and acts as a common dimerization partner for bHLH-PAS proteins during embryogenesis. A transient transfection assay using Drosophila S2 tissue culture cells and wild-type and mutant Drosophila tango cDNAs was used to localize the activation domain of the Tango protein. An activation domain was identified in the C-terminus of TGO consisting of poly-glutamine and histidine proline repeats. Transcriptional activation of the fibroblast growth factor receptor (breathless) gene required an intact TGO C-terminus, in vitro. Co expression assays of trachealess and tgo in the developing eye imaginal disc showed a requirement for the C-terminal transactivation domain of TGO for a cellular response. Genetic analysis of tgo(3) shows that the paired repeat is necessary for tracheal tubule formation in all branches. Lastly, expression of a C-terminal truncated tgo transgene specifically in the CNS midline and trachea resulted in reductions in the number of breathless-expressing cells. These results together identify TGO's transactivation domain and establish its importance for proper target gene regulation and cellular specification. PMID- 15818485 TI - Characterisation of Fmrp in zebrafish: evolutionary dynamics of the fmr1 gene. AB - Fragile X syndrome is the most common inherited form of mental retardation. It is caused by the lack of the Fragile X Mental Retardation Protein (FMRP), which is encoded by the FMR1 gene. Although Fmr1 knockout mice display some characteristics also found in fragile X patients, it is a complex animal model to study brain abnormalities, especially during early embryonic development. Interestingly, the ortholog of the FMR1 gene has been identified not only in mouse, but also in zebrafish (Danio rerio). In this study, an amino acid sequence comparison of FMRP orthologs was performed to determine the similar regions of FMRP between several species, including human, mouse, frog, fruitfly and zebrafish. Further characterisation of Fmrp has been performed in both adults and embryos of zebrafish using immunohistochemistry and western blotting with specific antibodies raised against zebrafish Fmrp. We have demonstrated a strong Fmrp expression in neurons of the brain and only a very weak expression in the testis. In brain tissue, a different distribution of the isoforms of Fmrp, compared to human and mouse brain tissue, was shown using western blot analysis. Due to the high similarity between zebrafish Fmrp and human FMRP and their similar expression pattern, the zebrafish has great potential as a complementary animal model to study the pathogenesis of the fragile X syndrome, especially during embryonic development. PMID- 15818486 TI - A validation study of the WHO analgesic ladder: a two-step vs three-step strategy. AB - GOALS OF WORK: The aims of the present study were to verify whether an innovative therapeutic strategy for the treatment of mild-moderate chronic cancer pain, passing directly from step I to step III of the WHO analgesic ladder, is more effective than the traditional three-step strategy and to evaluate the tolerability and therapeutic index in both strategies. METHODS: Patients aged 18 years or older with multiple viscera or bone metastases or with locally advanced disease were randomized. Pain intensity was assessed using a 0-10 numerical rating scale based on four questions selected from the validated Italian version of the Brief Pain Inventory. Treatment-specific variables and other symptoms were recorded at baseline up to a maximum follow-up of 90 days per patient. RESULTS: Fifty-four patients were randomized onto the study, and pain intensity was assessed over a period of 2,649 days. The innovative treatment presented a statistically significant advantage over the traditional strategy in terms of the percentage of days with worst pain > or =5 (22.8 vs 28.6%, p < 0.001) and > or =7 (8.6 vs 11.2%, p = 0.023). Grades 3 and 4 anorexia and constipation were more frequently reported in the innovative strategy arm, although prophylactic laxative therapy was used less in this setting. CONCLUSIONS: Our preliminary data would seem to suggest that a direct move to the third step of the WHO analgesic ladder is feasible and could reduce some pain scores but also requires careful management of side effects. PMID- 15818487 TI - [Main symptoms of acute abdomen]. PMID- 15818488 TI - Quantification of phase synchronization phenomena and their importance for verbal memory processes. AB - In the past years, interest in brain oscillations and their possible role in perceptual and cognitive processes has greatly increased. The two oscillations that have received the most attention are the theta and the gamma rhythm. In this study, the functioning and properties of phase synchronization parameters for these two frequency bands estimated by means of Gabor expansion were demonstrated with simulations for the phase-locking index (PLI) and the 1:1 as well as n:m phase synchronization indices. In order to demonstrate the importance of phase synchronization phenomena for memory performance, power, PLI and the 1:1 as well as n:m phase synchronization indices were calculated for EEG data on verbal memory encoding. These parameters showed various dissociations for recalled versus not-recalled nouns. In particular, the calculation of phase synchronization among different frequencies either at the same electrode or at different electrodes provided a completely new picture of dynamic neuronal interaction accompanying memory processing. PMID- 15818489 TI - Influence of methyl jasmonate on podophyllotoxin and 6-methoxypodophyllotoxin accumulation in Linum album cell suspension cultures. AB - The accumulation of podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (6MPTOX) was enhanced about twofold in the suspension culture of Linum album line 2-5 aH following the addition of methyl jasmonate (MeJas) to the cultivation medium, reaching 7.69+/-1.45 mg/g dry weight and 1.11+/-0.09 mg/g dry weight, respectively. There was no increase in 6MPTOX accumulation following the addition of MeJas to suspension cells of L. album line X4SF, whereas PTOX accumulation was enhanced about tenfold to 0.49+/-0.10 mg/g dry weight. Phenylalanine ammonia lyase activity increased immediately after the addition of MeJas to a cell suspension culture of line X4SF, reaching a maximum between 4 h and 1 day after elicitation, while cinnamyl alcohol dehydrogenase activity and the lignin content of the cells were not affected. PMID- 15818491 TI - Dispersed plug flow model for upflow anaerobic sludge bed reactors with focus on granular sludge dynamics. AB - A new approach to model upflow anaerobic sludge bed (UASB)-reactors, referred to as a one-dimensional dispersed plug flow model, was developed. This model focusses on the granular sludge dynamics along the reactor height, based on the balance between dispersion, sedimentation and convection using one-dimensional (with regard to reactor height) equations. A universal description of both the fluid hydrodynamics and granular sludge dynamics was elaborated by applying known physical laws and empirical relations derived from experimental observations. In addition, the developed model includes: (1) multiple-reaction stoichiometry, (2) microbial growth kinetics, (3) equilibrium chemistry in the liquid phase, (4) major solid-liquid-gas interactions, and (5) material balances for dissolved and solid components along the reactor height. The integrated model has been validated with a set of experimental data on the start-up, operation performance, sludge dynamics, and solute intermediate concentration profiles of a UASB reactor treating cheese whey [Yan et al. (1989) Biol Wastes 27:289-305; Yan et al. (1993) Biotechnol Bioeng 41:700-706]. A sensitivity analysis of the model, performed with regard to the seed sludge characteristics and the key model parameters, showed that the output of the dispersed plug flow model was most influenced by the sludge settleability characteristics and the growth properties (especially mu(m)) of both protein-degrading bacteria and acetotrophic methanogens. PMID- 15818495 TI - The cytogenetics of preimplantation human development: insights provided by traditional and novel techniques. AB - Our understanding of the incidence and origin of chromosome abnormalities in human preimplantation embryos is very limited due to the necessary ethical constraints involved in studying such material and the limited data ultimately produced. Several studies have addressed this issue, however, using techniques such as interphase fluorescence in situ hybridisation, modified G-banding preparation and the use of single-cell comparative genomic hybridisation (CGH). This review discusses the use of these techniques in assessing chromosome abnormalities in this, the earliest of human developmental stages. In addition, the prospects for the clinical use of CGH are discussed. PMID- 15818496 TI - Stimulus and response chunking in the Hebb Digits task. AB - Using the Hebb Digits task, an incidental sequential learning paradigm, the effects of chunking of both the presentation and response phases of performance were examined. In the first experiment, consistent stimulus chunking increased learning, and performance was at an equivalent level to this when consistent chunking of both stimuli and responses was present. Consistent chunking of the responses alone did not significantly improve learning over a baseline condition where neither stimuli nor responses were chunked. The disruption of response organization in a second experiment, through a random response condition, failed to impact learning in non-chunked and stimulus chunked conditions. A third experiment found that response chunking did benefit learning in a condition where stimuli were presented in random chunks. A final experiment suggested extended processing of the digits could not account for performance gains in the stimulus chunking condition. Overall, the results suggest that the enhanced effects of chunking on learning were stimulus-driven rather than response-driven, except under conditions that constrained a consistent pattern of stimulus organization. PMID- 15818497 TI - Activity-dependent modulation: a non-linearity in the unilateral vestibulo-ocular reflex pathways. AB - It is well established that the vestibulo-ocular reflex (VOR) depends not only on sensory stimulation but also on the behavioral context associated with the stimulation. Recent modeling studies suggested that including a non-linearity in the activation function of the VOR neurons achieves the desired context dependence for the VOR without resorting to currently assumed complex cortical computations. With the non-linearity, neurons operate as non-linear summers of incoming activity with sensitivities modulated by their activation levels. In this study we examined whether such a non-linearity exists in the unilateral VOR pathways in behaving monkeys. Acoustic clicks were employed to evoke unilateral VOR responses during fixation, head motion and smooth pursuit. We found that the click-evoked unilateral VOR responses did not simply sum in a linear manner with the eye movements initiated by head or target motion. Instead, the same acoustic click evoked larger eye movements if the ongoing eye movements were in the same direction. We also showed that the interaction between the ongoing eye movement and the click-evoked response was close to being multiplicative. These results revealed a previous unknown non-linearity in the unilateral VOR pathways, which may have important implications on the neural implementation of the context dependence for the VOR. PMID- 15818498 TI - Perceptual distortion of face deletion by local anaesthesia of the human lips and teeth. AB - As visual guidance of facial movements is impossible, accurate movements for speech and mastication require an established body image that is formed via the information from mechanoreceptors in the skin, mucosa, periodontium, and proprioceptors in the facial and masticatory muscles and in the jaw joints. In this study we aimed to investigate how the acute deafferentation of lips and teeth alters the established image of lips, teeth and the thumb. We used a psychophysical method to determine whether the perceived sizes of the upper lip and front teeth change when the sensory input from the lips and front teeth is fully blocked. We also examined the perceived size of the thumb to test for acute interactions between the thumb and facial structures. Local anaesthetic blocking of upper lip and upper front teeth significantly increased the perceived size of the upper lip by as much as 100% (range 21-100%) in ten out of eleven subjects tested (overall mean 52%; p=0.001). The perceived size of the upper teeth also significantly increased by as much as 155% (range 30-155%) in eight of the eleven subjects during anaesthesia (overall mean 41%; p=0.035). When the region of anaesthesia was increased and both upper and lower teeth and lips were anaesthetised, the perceived size of the upper lip again increased, by 53% (p=0.040), but the change in perceived size of the upper front teeth (18%) was not significant (p=0.206). In both studies there was no change in perceived size of the thumb. The results illustrate the labile central interaction between sensory inputs and the importance of feedback from peripheral afferents in generating the subjective facial image. The timing, level, and area of anaesthesia may be important modifiers of these interactions. PMID- 15818499 TI - Hand movement observation by individuals born without hands: phantom limb experience constrains visual limb perception. AB - Increasing evidence suggests that the visual analysis of other people's actions depends upon the observer's own body representation or schema. This raises the question of how differences in observers' body structure and schema impact their perception of human movement. We investigated the visual experiences of two persons born without arms, one with and the other without phantom sensations. These participants, plus six normally-limbed control observers, viewed depictions of upper limb movement under conditions of apparent motion. Consistent with previous results (Shiffrar M, Freyd JJ (1990) Psychol Sci 1:257), normally-limbed observers perceived rate-dependent paths of apparent human movement. Specifically, biologically impossible motion trajectories were reported at rapid display rates while biologically possible trajectories were reported at slow display rates. The aplasic individual with phantom experiences showed the same perceptual pattern as control participants, while the aplasic individual without phantom sensations did not. These preliminary results suggest that phantom experiences may constrain the visual analysis of the human body. These results further suggest that it may be time to move beyond the question of whether aplasic phantoms exist and instead focus on the question of why some people with limb aplasia experience phantom sensations while others do not. In this light, the current results suggest that somesthetic representations are not sufficient to define body schema. Instead, neural systems matching action observation, action execution and motor imagery likely contribute to the definition of body schema in profound ways. Additional research with aplasic individuals, having and lacking phantom sensations, is needed to resolve this issue. PMID- 15818500 TI - Effects of diazepam on the latency of saccades for luminance and binocular disparity defined stimuli. AB - Saccadic latency is composed of separate sensory and motor processing delays. Therefore, any alteration in the sensory processing should effect the saccadic latency. Because the highest density of benzodiazepine (Bz) binding sites is located in cerebral cortex, sensory processing of stimuli in this cortical area is expected to be substantially effected by administration of Bzs. It is well known that sensory processing of binocular disparity occurs in the cerebral cortical areas and therefore the latency of saccades to stimuli defined by binocular disparity should be substantially affected by Bz intake. In this study, we tested this prediction by comparing the latency of saccadic eye movements for binocular disparity defined stimuli (stereo stimuli) with those for luminance contrast defined stimuli (luminance stimuli), after diazepam or placebo. Eye movements were mainly recorded by use of the magnetic search coil technique, and the study was performed in a randomized, double-blind way. Although diazepam prolonged the latency of saccades for stereo and luminance stimuli, the percentage increases in saccadic latency for the stereo stimuli were significantly larger than those for the luminance stimuli. Saccadic peak velocity, and saccadic amplitude, also significantly decreased after diazepam under conditions of stereo and luminance stimuli. However, there was no significant difference for either saccadic peak velocity or amplitude between the two types of target. The results suggest that the latency of saccades to binocular disparity defined random-dot stimuli could more sensitively reflect the pharmacodynamic effects of Bzs on the cerebral cortex. PMID- 15818501 TI - Saccadic adaptation shifts the pre-saccadic attention focus. AB - The well-documented phenomenon of the spatial coupling between saccadic programming and the orienting of attention refers to the fact that visual attention is directed toward the location that the eyes are aiming for. However, the question remains open as to whether saccades and attention are two independent processes that can be directed concurrently toward a common goal, or whether their relationship is tighter, with the motor components of the saccade program influencing the selection of the position towards which visual attention is directed. To investigate this issue, an experiment was carried out in which the initial saccade goal was dissociated from the final executed motor vector. This was done by using a saccadic adaptation paradigm and a discrimination task. Results showed that best perceptual performance, which is taken to be an indicator of the locus of visual attention, followed the motor modifications arising from saccadic adaptation. This suggests that visual attention is directed toward the actual saccade landing position and that the perceptual system must have access to information regarding the motor vector before saccade execution. PMID- 15818502 TI - Protective effect of beta-glucan on lung injury after cecal ligation and puncture in rats. AB - OBJECTIVE: Understanding the biological mediators involved in the complex inflammatory response of sepsis and acute lung injury offers the possibility of future investigations targeting treatment based on these mediators. This study investigated whether macrophage activator beta-glucan has a protective effect on acute lung injury in an experimental model of sepsis. DESIGN AND SETTING: Experimental study in an experimental research center. MATERIALS: 30 rats randomized into three groups (sham, sepsis, and beta-glucan). INTERVENTIONS: Cecal ligation and puncture were performed in the beta-glucan and sepsis groups. The beta-glucan group was given a single intraperitoneal dose of beta-glucan (4 mg/kg) following cecal ligation. MEASUREMENTS AND RESULTS: Rats treated with beta glucan had fewer circulating neutrophils, more blood monocytes, and higher serum interleukin 6 levels than septic animals. The percentages of neutrophils and lymphocytes from the bronchoalveolar lavage fluid and the myeloperoxidase activity measured in the lung tissue were lower in the beta-glucan group than in the sepsis group. Less alveolar hemorrhage and neutrophil infiltration were observed in lungs from animals in the beta-glucan group in the septic groups. CONCLUSIONS: In this rat model of intra-abdominal sepsis beta-glucan treatment partially protected against secondary lung injury, decreased lung hemorrhages, and lung neutrophilia. These results suggest that beta-glucan protects against sepsis-associated lung damage. PMID- 15818503 TI - Enzymes as occupational and environmental respiratory sensitisers. AB - A literature review shows that airborne enzymes occurring in the general environment and in purified form in industrial production have a high allergenic potential to the airways, causing rhinitis, conjunctivitis and asthma. It can be assumed that this also applies to the increasing number of enzymes manufactured by the cloning of fast-growing genetically engineered microorganisms. Cross sectional studies demonstrate exposure-response relations for IgE-mediated sensitisation and airway disorders. Atopic individuals are more susceptible to enzyme allergy than non-atopic individuals. Skin prick testing and measurement of specific IgE antibodies have been shown to be useful diagnostic tools. Very high concentrations of proteases may lead to emphysema. There is also evidence for non allergic airway inflammation by proteases, probably via protease-activated receptor-2 and intracellular Ca(2+) release. It is recommended that all enzymes be classified with the risk phrase R42 (may cause sensitisation by inhalation) and that their inhalative uptake be totally avoided. PMID- 15818505 TI - Induction of tumor immunity and cytotoxic t lymphocyte responses using dendritic cells transduced by adenoviral vectors encoding HBsAg: comparison to protein immunization. AB - Dendritic cells (DC) are specialized antigen-presenting cells with powerful immunostimulatory properties. Their use for induction of anti-tumor immunity has been limited by several factors, including identification of appropriate tumor associated antigens, delivery of antigens to DC, and maintaining DC in a highly activated state. Here, DC propagated in vitro were transduced with an adenoviral (Ad) vector to express hepatitis B surface antigen (HBsAg), an antigen present in hepatocellular carcinoma (HCC). Many patients with HCC demonstrate evidence of prior HBV exposure, suggesting that the presence of the virus in a quiescent state may promote tumorigenesis. Ad-HBsAg-transduced DC stimulated strong cytotoxic T lymphocyte (CTL) responses to HBsAg-expressing tumor cells, and protected mice from lethal tumor challenge. Immunity was antigen-specific, as wild-type tumor (HBsAg -) grew normally. Furthermore, DC transduced with an irrelevant vector had no effect. Vaccination with HBsAg protein, a clinically utilized preparation that confers immunity to HBV infection, did not protect against tumor challenge even though it induced a strong antibody response. These studies describe for the first time the contributions of humoral and cellular immune responses to tumor immunity induced by Ad-transduced DC compared to protein vaccination. PMID- 15818506 TI - [Correct diagnosis for indication in gastroesophageal reflux disease]. AB - For diagnosis and quantitation of gastroesophageal reflux disease, a number of different examinations are available. The simplest are the patient's history and complaints as well as the proton pump inhibitor test; both methods have a positive predictive value of 65% to 70%. Esophagogastroduodenoscopy is the gold standard for differentiation between erosive and nonerosive reflux disease. Biopsy does not enhance the significance of endoscopy except for proof of Barrett's epithelium or malignant degeneration. Twenty-four-hour pH monitoring has the highest sensitivity and specificity for the diagnosis of gastroesophageal reflux disease. Barium swallow provides only additional information concerning hiatal hernia or stenosis. Manometry can clarify lower esophageal sphincter insufficiency or motility disorders of the tubular esophagus. However, the available studies show that manometry has no influence on postoperative outcome after fundoplication. The indication for medical therapy can primarily be based only on the history and complaints of the patient. As surgical therapy affords a high degree of diagnostic accuracy, at least endoscopy and 24-h pH monitoring are necessary for indication. PMID- 15818507 TI - Development of a pharmacokinetic limited sampling model for temozolomide and its active metabolite MTIC. AB - PURPOSE: To develop a pharmacokinetic limited sampling model (LSM) for temozolomide and its metabolite MTIC in infants and children. METHODS: LSMs consisting of either two or four samples were determined using a modification of the D-optimality algorithm. This accounted for prior distribution of temozolomide and MTIC pharmacokinetic parameters based on full pharmacokinetic sampling from 38 patients with 120 pharmacokinetic studies (dosage range 145-200 mg/m(2) per day orally). Accuracy and bias of each LSM were determined relative to the full sampling method. We also assessed the predictive performance of the LSMs using Monte-Carlo simulations. RESULTS: The four strategies generated from the D optimality algorithm were as follows: LSM 1=0.25, 1.25, and 3 h; LSM 2=0.25, 1.25, and 6 h; LSM 3=0.25, 0.5, 1.25, and 3 h; LSM 4=0.25, 0.5, 1.25, and 6 h. LSM 2 demonstrated the best combination of low bias [0.1% (-8.9%, 11%) and 11% (4.3%, 15%)] and high accuracy [-1.0% (-12%, 24%) and 14% (7.9%, 37%)] for temozolomide clearance and MTIC AUC, respectively. Furthermore, adding a fourth sample (e.g., LSM 4) did not substantially decrease the bias or increase the accuracy for temozolomide clearance or MTIC AUC. Results from Monte-Carlo simulations also revealed that LSM 2 had the best combination of lowest bias (0.1+/-6.1% and -0.8+/-6.5%), and the highest accuracy (4.5+/-4.1% and 5.0+/ 4.3%) for temozolomide clearance and MTIC apparent clearance, respectively. CONCLUSIONS: Using data derived from our population analysis, the sampling times for a limited sample pharmacokinetic model for temozolomide and MTIC in children are prior to the temozolomide dose, and 15 min, 1.25 h and 6 h after the dose. PMID- 15818508 TI - A phase I, dose-finding study of irinotecan (CPT-11) short i.v. infusion combined with fixed dose of 5-fluorouracil (5-FU) protracted i.v. infusion in adult patients with advanced solid tumours. AB - PURPOSE: Irinotecan (CPT-11) and 5-fluorouracil (5-FU) are effective cytotoxic agents in the treatment of solid tumours. Continuous i.v. infusion (CI) of 5-FU is significantly more active and better tolerated than bolus i.v. 5-FU. This phase I pharmacokinetic and clinical study evaluated escalating CPT-11 doses administered every 3 weeks combined with a fixed dose of 5-FU CI over 14 days to find the maximum tolerated dose (MTD) of this combined chemotherapy. PATIENTS AND METHODS: Patients with solid tumours showing failure with previous standard treatment or for whom no established curative therapy existed received CPT-11 i.v. over 90 min (six dose levels were evaluated: 150, 175, 200, 250, 300 and 350 mg/m(2)) plus a fixed dose of 5-FU CI 250 mg/m(2) per day over 14 days. If the MTD was not reached at CPT-11 level 6, then 5-FU was increased to 300 mg/m(2). In step 2, 5-FU was administered as a true protracted infusion at the recommended dose found during step 1. In step 3, the recommended dose of CPT-11 was divided and administered in a weekly schedule for 4 weeks combined with a fixed dose of 5 FU CI 250 mg/m(2), and then followed by 2-5 weeks rest. RESULTS: Neutropenia and diarrhoea were the main toxicities, leading to early termination of infusion in three of six patients in level 7. Therefore, CPT-11 350 mg/m(2) + 5-FU 250 mg/m(2) CI over 14 days was identified as the recommended dose. In step 2, CPT-11 dose had to be reduced to 300 mg/m(2) due to toxicity. The weekly schedule of CPT 11 75 mg/m(2) + 5-FU 250 mg/m(2) CI was feasible with only one patient experiencing severe diarrhoea. No interactions were found in the kinetics parameters of CPT-11 or 5-FU for the different dose levels studied. CONCLUSION: CPT-11 300 mg/m(2) + 5-FU 250 mg/m(2) protracted infusion is the recommended dose for phase II trials, neutropenia and diarrhoea being the dose-limiting toxicities. PMID- 15818509 TI - Histone hypoacetylation is involved in 1,10-phenanthroline-Cu2+-induced human hepatoma cell apoptosis. AB - The 1,10-orthophenanthroline (OP)-Cu(2+) combination, one generally used reactive oxygen species (ROS) generation system, is known to induce cell apoptosis, but the mechanism of ROS generation in this process remains unclear. Here we found that in the presence of 5 microM Cu(2+), OP inhibited histone acetyltransferase (HAT) activity, resulting in decreased acetylation in both histone H3 and H4. This inhibition of histone acetylation and HAT activity was significantly attenuated by preventing or scavenging ROS generation with the Cu(2+) chelator of bathocuproine disulfonate, or the antioxidants of N-acetyl-cysteine and mannitol, respectively, indicating the involvement of ROS generation in OP-Cu(2+) -induced histone hypoacetylation. At the same time, this ROS generation is found to be involved in OP-Cu(2+) -induced apoptosis in human hepatoma Hep3B cells. The important role of histone hypoacetylation in the induction of apoptosis was also proven by the marked diminution of apoptosis by 100 nM trichostatin A, a specific inhibitor of histone deacetylase, or the overexpression of p300, an HAT protein. Collectively, these observations suggest that histone hypoacetylation represents one unrevealed mechanism involved in the in vivo function of OP-Cu(2+) -generated ROS, at least in their induction of cell apoptosis. PMID- 15818512 TI - Takayasu's arteritis and chronic autoimmune thyroiditis in a patient with type 1 diabetes mellitus. AB - Takayasu's arteritis is a rare, chronic inflammatory disease of unknown origin, affecting the walls of the aorta and its main branches, as well as coronary and pulmonary arteries. The inflammation of the arteries may lead to stenosis, occlusions, dilatations, and aneurysms of involved vessels. It is relatively common in Asia and the Far East but is rare in the Western Hemisphere. We present the case of a 36-year-old white woman with a history of type 1 diabetes mellitus and chronic autoimmune thyroiditis who complained of easy fatigability in the upper limbs, with absent arterial pulses in the upper limbs and audible bruits over both subclavian and left common carotid arteries. Intra-arterial digital subtraction angiography revealed complete or subtotal obliteration of the aortic arch's branches, with the brain supplied with blood only by the left vertebral artery originating directly from the aortic arch. We diagnosed Takayasu's arteritis with abnormal origin of the left vertebral artery. To the best of our knowledge, our case of Takayasu's arteritis and chronic autoimmune thyroiditis in a type 1 diabetic patient with abnormal origin of the left vertebral artery is the first one ever described. PMID- 15818511 TI - Lymphocyte apoptosis and macrophage function: correlation with disease activity in systemic lupus erythematosus. AB - Increased lymphocyte apoptosis and defects in macrophage removal of apoptotic cells have been suggested to contribute to the development of systemic lupus erythematosus (SLE). The aim of this study was to investigate the relationship between peripheral lymphocyte apoptosis, macrophage function as determined by the serum levels of neopterin and interferon-gamma (IFN-gamma), and SLE disease activity. Peripheral apoptotic lymphocytes (AL) were detected by annexin V fluorescein isothiocyanate (FITC) staining and flow cytometry. Serum levels of neopterin and IFN-gamma were measured by enzyme-linked immunosorbent assay (ELISA). SLE disease activity was determined using the systemic lupus activity measure (SLAM) and the serum titer of anti-dsDNA antibodies. The percentage of AL in the peripheral blood of active SLE patients was significantly higher (13.07+/ 7.39%, n=30) than that of the inactive SLE patients (4.08+/-3.55%, n=8, p<0.01) and normal controls (5.13+/-3.37%, n=11, p<0.01). Serum levels of neopterin in active SLE patients were significantly higher (1.39+/-1.10 microg/dl, n=22) than in controls (0.26+/-0.19 microg/dl, n=20, p<0.01). Serum levels of IFN-gamma in active SLE patients were elevated (58.97+/-34.52 ng/l, n=15) when compared with controls (28.06+/-2.35 ng/l, n=16, p<0.05). The percentage of AL correlated significantly with serum levels of neopterin (r=0.446, p<0.05, n=22) and SLAM score (r=0.533, p<0.001, n=38), but not with the serum levels of IFN-gamma. The SLAM score also correlated with the serum levels of neopterin (r=0.485, p<0.05, n=22), but not with those of IFN-gamma. Our study supported the hypothesis that increased lymphocyte apoptosis has a pathogenic role in SLE. The increased levels of serum neopterin may suggest an attempt of the patients' macrophage system to remove the apoptotic cell excess. Since serum levels of neopterin correlated with the overall lupus disease activity, they may be regarded as an index of SLE disease activity. PMID- 15818513 TI - Marked increase of serum hepatitis C virus (HCV) RNA titer during treatment with high-dose prednisolone in a case of polymyositis. AB - A 70-year-old Japanese woman with hepatitis C virus (HCV) infection was diagnosed with polymyositis and treated with high-dose prednisolone (PSL). The serum alanine aminotransferase (ALT) level increased from 78 to 345 U/l 1 week after initiating treatment, although the polymyositis settled promptly. Furthermore, the serum HCV RNA level increased markedly from 110 to 850 kIU/ml 3 weeks after starting treatment. Previously, the patient had suffered an occlusion of the left branch of the retinal vein secondary to hyperviscosity syndrome resulting from Sjogren's syndrome and low-dose PSL treatment had been commenced. The serum HCV RNA and transaminase levels had not increased during this low-dose PSL treatment. Although intensive immunosuppression is necessary as an initial treatment of several collagen diseases including polymyositis, high-dose PSL therapy may markedly augment the serum HCV RNA level and therefore careful observation is necessary in HCV-infected patients. PMID- 15818514 TI - [Motor disturbances in psychosis]. PMID- 15818515 TI - [Motor disturbances in psychiatric patients]. AB - Motor disturbances are common symptoms in schizophrenia. They can be caused by the illness itself or by antipsychotic treatment. The author gives an overview of the history of research in this topic and discusses the relationship of motor disturbances with many other aspects of psychopathology and subjective well-being of the patients, such as cognitive, emotional and social functioning. PMID- 15818516 TI - [Catatonia]. AB - Catatonia is a neuropsychiatric syndrome characterized by mental, motor and behavioral symptoms. It occurs in up to 18 % of acute admissions and is most frequently associated with affective and psychotic disorders. It is also seen in dissociative disorders, pervasive developmental disorders, mental retardation and organic psychiatric disorders. Catatonic syndromes are impressive states that can be reliably and validly diagnosed both clinically and with psychometric measurements and they can be treated effectively. Despite this, they are often not recognized in clinical practice, are not part of the therapeutic strategy and thus remain untreated. The following article is intended to give a review of the most pertinent questions related to the diagnosis and treatment of catatonia in order to improve clinicians' ability to recognize and treat catatonic symptoms and syndromes adequately. PMID- 15818517 TI - [Extrapyramidal side-effects: avoidance, diagnosis and treatment]. AB - Acute extrapyramidal side-effects (EPS) are a common phenomenon of treatment with antipsychotics. They are associated with a substantial reduction of the patient's quality of life and of compliance with the treatment. This article gives an overview of the different forms of EPS. It outlines strategies to avoid these side-effects and discusses problems in diagnosis and treatment. In comparison to conventional compounds, the clearest advantage of new generation antipsychotics is a reduced risk of the occurrence of extrapyramidal side-effects (EPS). Many studies have found that newer antipsychotics have a lesser propensity to causes EPS. Nevertheless, there are some conflicting results on this topic. This article discusses some of these results and sheds light on methodological problems in the evaluation of side-effects of antipsychotic treatment. PMID- 15818518 TI - [Kinematic movement analyses and their application in psychiatry]. AB - There is a long tradition to develop valid instruments for the exact assessment of psycho-motor dysfunctions in psychiatry. However, progress is hampered by the complexity of emotionally driven movements in psychiatric patients. Methods used up to now either remains unspecific due to only qualitative measurements or focus on the neurophysiological aspects too much. Thus, the results accomplished so far are only very general unspecific concerning different groups of psychiatric patients. In this paper, two own methods are presented which are aimed to avoid the two poles above mentioned. Kinematic analyses of facial expressions as well as handwriting movements provide quantitative and quite specific informations about psycho-motor dysfunctions of psychiatric patients and the effects of psychotropic substances. Thus, these methods are well suitable for relating them to other neurobiological parameters in order to contribute to the pathophysiological understanding of psycho-motor symptoms in psychiatric patients. PMID- 15818519 TI - [Three-dimensional ultrasonic movement analysis of gait and hand-movements in schizophrenic patients]. AB - Neurological signs and disturbed psychomotor performance are consistently confirmed by clinical studies on schizophrenic patients. They could either be caused by the illness itself or could occur as side-effects of antipsychotic medication. Based on the clinical observation, differential diagnosis between catatonic symptoms of schizophrenia and extrapyramidal side-effects often remains unclear. Apart from their diagnostic value, motor disturbances can also be predictive parameters for prognosis of the disease and clinical response to antipsychotic treatment. Motor disturbances are mainly assessed by use of clinical ratings. In the last years, new approaches like infrared or ultrasonic movement analysis systems have been introduced to objectively determine motor disturbances in schizophrenic patients. Ultrasonic movement analysis systems calculate the three-dimensional positions of tiny markers, which are attached to moving body parts, with high spatial and temporal resolution. Thus, key parameters of gait and hand-movements can be determined exactly. Results of several studies using these new methods indicate that schizophrenia causes a primary disturbance of motor performance. Treatment with conventional antipsychotics intensifies many of the disabling motor deficiencies, whereas the influence of atypical antipsychotics on motor performance is either not detectable or much smaller. All in all, most results of studies using objective methods for the assessment of motor performance underline the advantages of treatment with atypical antipsychotics compared to conventional antipsychotics, particularly with regard to the patient's motor performance. PMID- 15818520 TI - [The neuronal level of motor activity: determination of motor cortex excitability by TMS]. AB - Transcranial magnetic stimulation as mapping method offers the possibility to measure aspects of motor cortex excitability painlessly and non-invasively. Using this neurophysiological tool, new insights into the effects of central-acting drugs are possible. Particularly striking seems to be the potential of this approach to gain new insights into neurobiological processes associated with neuropsychiatric diseases like schizophrenia or major depression. In combination with genetic aspects, TMS is able to bridge the gap between molecular research and clinical approach. PMID- 15818521 TI - [Functional magnetic resonance tomography of central motor areas in schizophrenic patients]. AB - Functional Magnetic Resonance Tomography (FMRT) is a safe and noninvasive method to study human brain function by measuring task induced changes of the regional cerebral blood flow. Recent fMRT studies showed that stimulation method, psychiatric disorders and neuroleptic agents influence motor induced brain activation patterns specifically. Since these factors have been insufficiently controlled in most of the recent fMRT studies in particular on psychiatric disorders, results are often controversial. Using an intrapersonal study design and a standardized stimulation apparatus, we explored differential effects of typical and atypical neuroleptic treatment in schizophrenia. In addition to fMRT, 3D ultrasonic assisted motor analysis was used to investigate gait and hand movements. FMRT method, fMRT studies on human motor system, fMRT data on schizophrenia as well as on the influence of neuroleptic treatment are given. In particular, the own study design is presented and preliminary data are given. PMID- 15818522 TI - [On the importance of care for the mentally ill and options for legal health care insurance]. PMID- 15818523 TI - [Political decisions and implementation: efficient management of a psychiatric department be successful despite working time law, practice fee and other innovations?]. PMID- 15818524 TI - [Symposium on the measurement of the quality of life in medicine and health ecology on December 3rd 2004 in Leipzig]. PMID- 15818526 TI - [Quo Vadis? Health policy decisions and their consequences for mental health care]. PMID- 15818527 TI - [The direct costs of depressive disorders in Germany]. AB - OBJECTIVE: The purpose of this study was to estimate the direct costs of depression in Germany from the social perspective. METHODS: The data came from the European Study of the Epidemiology of Mental Disorders (ESEMeD), in which a representative sample of 3555 non-institutionalised adults aged 18 years or older was interviewed in Germany. Using the most recent version of the Composite International Diagnostic Interview (WMH-CIDI), 131 persons with a 12-month prevalence of major depression, minor depression and dysthymia were identified. These patients reported their mental health care consumption of the last 12 months retrospectively, which was valued by prices of 2002. RESULTS: The average direct costs for treating depressive disorders were 686 EUR per patient and year. The total direct costs of depression in Germany was estimated at approximately 1.6 billion EUR. CONCLUSIONS: Because the sample was restricted to the non institutionalised population, calculated costs may be underestimated. The costs per inhabitant in Germany exceed those reported by studies from the United Kingdom but are lower than those reported for the USA. PMID- 15818528 TI - [Measuring preferences for depressive health states]. AB - OBJECTIVE: The purpose of this study was to analyse the construct validity and the concurrent validity of the EuroQol instrument (EQ-5D self-classifier, EQ VAS and EQ-5D indices), time trade-off (TTO) and contingent valuation (willingness-to pay) for preference-based valuation of depressive health states. METHODS: A survey of a representative sample (N = 3522) of the non-institutionalised population aged 18 and above in Germany was conducted using the EQ-5D, TTO and contingent valuation as well as the SF-12 questionnaire. Construct validity was analysed by comparing 130 respondents in whom 12-month prevalence for major depression, minor depression, or dystimia according to DSM-IV was identified based on a version of the Composite International Diagnostic Interview (WMH-CIDI) to respondents without depressive disorders. Concurrent validity was analysed by assessing the correlation between these measures in the group with depressive disorders. RESULTS: In the group with depressive disorders, problems in all EuroQol dimension except for mobility were significantly more frequent, and mean values of all analysed preference measures except for willingness-to-pay were less favourable than in the group without depressive disorders (EQ VAS score 67 vs. 78; EQ-5D index [VAS] 83 vs. 94; EQ-5D index [TTO] 0.81 vs. 0.91; TTO 0.89 vs. 0.95; always p < 0.001). In the group with depressive disorders, EQ VAS score and EQ-5D indices were significantly correlated one with another (ranging from r = 0.63 to r = 0.92) as well as with the mental and the physical component scale of the SF-12 (ranging from r = 0.26 to r = 0.45) and with willingness-to-pay (ranging from r = - 0.20 to r = - 0.22); besides, willingness-to-pay was significantly correlated with TTO (r = - 0.46). CONCLUSIONS: The results of this study point to the construct validity and concurrent validity of preference measures in patients with depressive disorders. PMID- 15818529 TI - [Direct costs of acute day hospital care: results from a randomized controlled trial]. AB - OBJECTIVE: So far only five randomized controlled trials on acute day hospital care have assessed direct health care costs and compared these with costs of conventional inpatient treatment. This paper aims to close this research gap for German speaking countries. METHOD: Another trial was conducted at the Department of Psychiatry and Psychotherapy, Dresden University of Technology. Using the Client Service Receipt Inventory interviews with patients at three time-points (admission, discharge, three months after discharge) assessed their utilization of health care services. Based on the results of a separate regional cost calculation project, costs of individual care packages could be calculated. 144 patients (day care: n = 75, inpatient care: n = 69) with complete data sets for all time-points of assessment were included in the intention-to-treat-analysis. Means of direct costs (given in Euro and referred to Deutsche Mark prices in 2000) were computed, and t-tests and bootstrap-procedures were used for group comparison. Furthermore, missing and sensitivity analyses were conducted. RESULTS: Patients in the acute day hospital caused mean direct health care costs of 12 401 per person within the entire period assessed. Thus, their cost level falls below the mean costs of inpatient care (15,924 euro per person) by 22.1 %. Missing analyses showed no selection effects on cost results caused by patients who could not be assessed at all defined time-points. Costs for inpatient and day care services were the most sensitive parameters for maintaining the statistically significant differences of cost means demonstrated between the two study groups. CONCLUSION: For German-speaking countries, this study shows for the first time that acute day care -- which has been demonstrated to be clinically at least as effective as inpatient care - is the less expensive option if these two settings are comparatively assessed. PMID- 15818530 TI - [Cost-effectiveness of galantamine in a german context]. AB - OBJECTIVE: The application of acetylcholinesterase (AChE) inhibitors seems to be constrained in the German health care system. We investigate the cost effectiveness of the AChE inhibitor Galantamine for patients with Alzheimer's disease (AD) in a German context. METHOD: A Markov model as the decision theoretic framework is applied to compare two treatment scenarios (Placebo vs. Galantamine). The incremental cost-utility ratio based on different sources of data is calculated as decision criteria. RESULTS: In the base case, the application of the AChE inhibitor Galantamine is a dominant scenario with cost savings along with gained quality adjusted life years. It is not cost saving from the perspective of third party payers in Germany. Results are sensitive regarding assumptions about efficacy of the drug and mortality in the analysed cohort. CONCLUSION: The German reimbursement system places an obstacle to the application of Galantamine. Further research is needed to tackle uncertainties in the model. PMID- 15818531 TI - In silico analysis of disease-association mapping strategies using the coalescent process and incorporating ascertainment and selection. AB - We present a new method for simulating samples of marker haplotypes, genotypes, or diplotypes in case-control studies in which the markers are linked to a disease locus in any specified region of the genome. The method allows realistic features to be incorporated into the simulations, including selection acting on disease alleles, sample ascertainment of disease chromosomes and polymorphic markers, a genetic dominance model of disease expression that allows incomplete penetrance and phenocopies, and an accurate genetic map of recombination rates and hotspots for recombination in the human genome (or, alternatively, an improved method for simulating the distribution of hotspots). The new method uses an approach that combines simulation of the coalescent process for the sampled chromosomes with a diffusion process used to model the evolution of the disease mutation frequency over time. Examples illustrate how the method may be used to study the expected power of a marker-disease association study. PMID- 15818532 TI - Pemetrexed (Alimta) in small cell lung cancer. AB - Small cell lung cancer (SCLC) comprises approximately 13% of all lung cancers. In limited stage (LS)-SCLC, combined-modality therapy represents the standard of care. Therapy should be approached curatively in fit patients with a good performance status because 5-year survival rates approach 26% in aggressively treated patients. In contrast, cure is not possible in extensive stage (ES)-SCLC with median 2-year survival rates with current therapy remaining at less than 10%. Pemetrexed (Alimta; Eli Lilly and Co, Indianapolis, IN) is a novel, multi targeted antifolate that inhibits several folate-dependent enzymes involved in purine and pyrimidine synthesis, and is active as a single-agent or in combination with a platinum in both non-small cell lung cancer and malignant pleural mesothelioma. Pemetrexed/platinum combinations appear active in ES-SCLC based on objective response rates observed in a randomized phase II trial. However, no survival data is yet available from this trial. The toxicity profile of both cisplatin and carboplatin in combination with pemetrexed was extremely favorable, as was the ability to deliver full doses of each of the component drugs. Given the limited options available for patients in the relapsed setting, the activity of single-agent pemetrexed is interesting. Also, preliminary data indicates that full doses of carboplatin/pemetrexed can be administered with thoracic radiation therapy, supporting a future clinical trial initiative in LS SCLC. PMID- 15818533 TI - The evolving role of pemetrexed (Alimta) in lung cancer. AB - Pemetrexed (Alimta; Eli Lilly and Co, Indianapolis, IN) is a multitargeted antifolate that inhibits several folate-dependent enzymes that play roles in purine and pyrimidine synthesis. The principal toxicities of pemetrexed are neutropenia, diarrhea, nausea/vomiting, mucositis, and skin rash. These toxicities are more frequent in vitamin-deficient (folate and vitamin B 12 ) patients, and can be ameliorated by the co-administration of folate and vitamin B 12 . The use of prophylactic dexamethasone is also recommended to reduce the frequency of severe skin rash. Pemetrexed has significant single-agent activity in previously treated and untreated patients with non-small cell lung cancer (NSCLC). A recent phase III trial comparing pemetrexed with docetaxel in previously treated NSCLC patients showed equivalent efficacy with less bone marrow toxicity (eg, neutropenia) in the pemetrexed group. These results were pivotal in the approval of pemetrexed for the treatment of refractory NSCLC. Pemetrexed has been combined with the platinums (ie, cisplatin, carboplatin, and oxaliplatin) in NSCLC to yield clinical activity similar to that of other platinum-based doublets. A comparative phase III trial of cisplatin/pemetrexed against cisplatin/gemcitabine (Gemzar; Eli Lilly and Co) is under way. Pemetrexed has also been evaluated in combination with gemcitabine, and although the optimal dose and schedule of this combination has not been defined, clinical activity similar to other nonplatinum-based doublets has been observed. Preliminary evidence suggests that pemetrexed can be combined with thoracic radiation therapy, but more data are needed to evaluate the potential advantage(s) pemetrexed may have in this setting. Pemetrexed/platinum doublets also appear to possess activity in extensive stage small cell lung cancer. A phase II trial of single-agent pemetrexed is under way in both sensitive- and refractory-relapsed small cell lung cancer. Given the activity and excellent tolerability of pemetrexed, further studies in lung cancer are warranted. PMID- 15818537 TI - Pemetrexed plus carboplatin or oxaliplatin in advanced non-small cell lung cancer. AB - Over the past decade many large randomized trials have been conducted in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC); however, no single regimen has shown clear superiority. Typical response rates of 17% to 32%, median survival times of 7 to 10 months, and 1-year survival rates of 30% to 45% are common in such studies. Nevertheless, metastatic NSCLC remains an incurable disease, with the development of drug resistance as a major impediment. New treatment regimens, such as the broadly used antifolate antimetabolites, target multiple metabolic functions and may lead to improvements in clinical outcome. Pemetrexed (Alimta; Eli Lilly and Co, Indianapolis, IN) is a novel antifolate, distinguished by a unique nuclear structure and its ability to inhibit multiple folate-dependent enzymatic pathways. Typically, vitamin B 12 and folic acid supplementation are incorporated into the treatment regimen to ameliorate untoward toxicities. When used either as a single agent or in combination with cis-diamine platinums (eg, cisplatin, carboplatin) or diaminocyclohexane platinums (eg, oxaliplatin), pemetrexed has shown significant antitumor activity, with mild, manageable toxicity in patients with advanced NSCLC. PMID- 15818539 TI - Oral water load and the 'Internal Tsunami'. PMID- 15818538 TI - Adjuvant and neoadjuvant therapy in non-small cell lung cancer. AB - The 5-year survival rates for patients with non-small cell lung cancer (NSCLC) ranges from 9% to 61% following resection, depending on clinical stage; survival rates post-surgery (pathologic stage) range from 25% to 67%. Most stage I and II patients eventually experience recurrent disease: two thirds occur systemically, one third locally. Surgical resection remains the standard of care in early stage NSCLC, although the role of surgery in stage IIIA [N 2 ] disease is controversial. Despite resection, the vast majority of lung cancer patients will experience recurrent and/or metastatic disease; therefore, supplementing surgery with adjuvant therapy is a rational treatment strategy. Recent data indicate that adjuvant chemotherapy should now be considered the standard of care for the treatment of patients with completely resected early stage NSCLC, with the single exception of patients with stage IA disease, where the prognosis is relatively favorable and there is currently no evidence supporting the efficacy of adjuvant therapy. While recent data from trials of adjuvant chemotherapy have shown promising results, no study has yet compared the utility of adjuvant versus neoadjuvant, or induction, chemotherapy. From the current data, more than 90% of patients receiving neoadjuvant chemotherapy undergo the planned surgical resection. Neoadjuvant chemotherapy may also downstage the disease before surgery and decrease perioperative tumor seeding, and molecularly targeted approaches with neoadjuvant therapy appear promising. PMID- 15818540 TI - Experimental evaluation of transport force in the rabbit ureter. AB - BACKGROUND: Cleaning the urinary tract by so-called 'wash-out effect' and promoting high diuresis has long been advocated but has had very little scientific backing and few prospective studies in international journals. AIM: To verify whether the physical laws describing the transport force of water in rivers and pipes are also valid for urinary outflow. METHODS: A laboratory model for measuring transport force, given liquid and solid capacity, was adapted to create an in vivo model based on the rabbit urinary tract. RESULTS: Fluid flow in the rabbit renal pelvis and ureters was found similar to flow in pipes, obeying the physical laws of water transport to some extent. When the quantity of liquid flowing in the urinary tract in unit time was doubled, the transport force increased by various orders of magnitude. When the liquid increased by a larger factor, the transport force became enormous. CONCLUSIONS: The results confirm the utility of maintaining high diuresis in patients with renal calculus, but stress the utility of drinking 1-2 liters of hypotonic water in a short time to obtain an enormous increase in transport force which increases the probability of a cleansing effect. PMID- 15818541 TI - A mathematical approach to the evaluation of sediment transport capacity in the human urinary tract. AB - BACKGROUND: It is not yet clear whether forced diuresis is useful for flushing out of the urinary tract, especially in cases of renal calculi or infections. A series of experiments conducted initially in vitro and then in vivo in rabbits, using sand and small glass spheres, showed that sediments can be eliminated from the urinary tract by forced diuresis. When distilled water was used, the effect was greater, both as weight of sand and as number of spheres eliminated, than when normal saline was used. The different characteristics of the two liquids gave rise to different intrapelvic pressures for a given flow. MATERIALS AND METHODS: Intrapelvic pressures, and liquid and solid discharges in vitro and in rabbits were used in a fluid mechanics analysis of the phenomenon to obtain non dimensional formulae so that the results in rabbits could be extended to humans. RESULTS: The results made it possible to evaluate solid transport capacity induced in a healthy human of given weight by drinking a given quantity of water in an hour. CONCLUSIONS: The mathematical model makes it possible to determine the quantity of water a patient must drink to induce a transport force sufficient for urinary tract wash-out. PMID- 15818542 TI - Hyperhomocysteinemia in hemodialysis patients: is it a risk factor for cardiovascular disease? PMID- 15818543 TI - Lamivudine in the treatment of HBV-related liver disease after renal transplantation: an update. AB - Diminished survival due to hepatitis B has been observed after renal transplantation (RT). Lamivudine, a second-generation nucleoside analogue, has been approved for the treatment of chronic hepatitis B virus (HBV) infection in patients with normal renal function. Numerous clinical experiences with lamivudine after RT have been recently published. Despite numerous shortcomings, all of these reports have shown encouraging results. The rate of clearance of HBV viremia ranged between 67% and 100%, and the frequency of ALT normalization was between 50% and 100% with lamivudine use. Even patients with fibrosing cholestatic hepatitis, a serious form of HBV-related liver disease with ominous course, have been successfully treated with lamivudine. Lamivudine therapy significantly improved the survival of HBsAg positive renal allograft recipients. However, numerous issues concerning the treatment of hepatitis B after RT remain unclear: the optimal time to initiate lamivudine, the appropriate duration of antiviral therapy after RT, and the role for pre-transplantation liver biopsy. Also, the management of lamivudine resistance remains a concern for physicians. Clinical trials are under way. PMID- 15818544 TI - Low-dose citrate continuous veno-venous hemofiltration (CVVH) and acid-base balance. AB - OBJECTIVE: To evaluate the acid-base effect of low-dose regional citrate anticoagulation (RCA) during continuous veno-venous hemofiltration (CVVH). DESIGN: Prospective observational study. SETTING: ICUs of tertiary public and private hospitals. SUBJECTS: Thirty critically ill patients with acute renal failure at risk of bleeding or with a major contraindication to heparin-CVVH and/or short filter life. METHODS: We used a commercial citrate-based fluid (11 mmol/L, sodium: 140 mmol/L, chloride: 108 mmol/L and 1 mol/L of potassium) as pre dilution replacement fluid during CVVH. Further potassium was added according to serum potassium levels. We measured all relevant variables for acid-base analysis according to the Stewart-Figge methodology. RESULTS: Before treatment, study patients had a slight metabolic acidosis, which worsened over 6 hours of RCA-CVVH (pH from 7.39 to 7.38, p < 0.005; bicarbonate from 23.2 to 21.6 mmol/L, p < 0.0001 and base excess from -2.0 to -3.0 mEq/L, p < 0.0001) due to a significant increase in SIG (from 5.8 to 6.6 mEq/L, p < 0.05) and a decrease in SIDa (from 37.5 to 36.6 mEq/L, p < 0.05). These acidifying effects were attenuated by hypoalbuminemia and a decrease in lactate (from 1.48 to 1.34 mmol/L, p < 0.005) and did not lead to progressive acidosis. On cessation of treatment, this acidifying effect rapidly self-corrected within six hours. CONCLUSIONS: Low dose RCA-CVVH induces a mild acidosis secondary to an increased strong ion gap and decreased SIDa which fully self-corrects at cessation of therapy. Clinicians need to be aware of these effects to correctly interpret changes in acid-base status in such patients. PMID- 15818545 TI - Serum potassium and acid-base parameters in severe dialysis-associated hyperglycemia treated with insulin therapy. AB - We analyzed the changes in serum potassium concentration ([K]) and acid-base parameters in 43 episodes of dialysis-associated hyperglycemia (serum glucose level > 33.3 mmol/L), 22 of which were characterized as diabetic ketoacidosis (DKA) and the remaining 21 as nonketotic hyperglycemia (NKH). All episodes were treated with insulin therapy only. Age, gender, initial and final serum values of glucose, sodium, chloride, tonicity and osmolality did not differ between DKA and NKH. At presentation, serum values of [K] (DKA 6.2 +/- 1.3 mmol/L; NKH 5.2 +/- 1.5 mmol/L) and anion gap [AG] (DKA 27.2 +/- 6.4 mEq/L; NKH 15.4 +/- 3.5 mEq/L) were higher in DKA, whereas serum total carbon dioxide content [TCO2 ] (DKA 12.0 +/- 4.6 mmol/L; NKH 22.5 +/- 3.1 mmol/L), arterial blood pH (DKA 7.15 +/- 0.09; NKH 7.43 +/- 0.07) and arterial blood PaCO2 (DKA 26.2 +/- 12.3 mm Hg; NKH 34.5 +/ 6.7 mm Hg) were higher in NKH. At the end of insulin treatment, serum values of [K] (DKA 4.0 +/- 0.7 mmol/L, NKH 4.0 +/- 0.5 mmol/L), [AG] (DKA 16.3 +/- 5.4 mEq/L, NKH 14.9 +/- 3.0 mEq/L), [TCO2 ] (DKA 23.5 +/- 5.0 mmol/L, NKH 24.1 +/- 4.2 mmol/L), arterial blood pH (DKA 7.42 +/- 0.09, NKH 7.51 +/- 0.14) and arterial blood PaCO2 (DKA 31.8 +/- 6.7 mm Hg, NKH 34.2 +/- 8.3 mm Hg) did not differ between the two groups. Linear regression of the decrease in serum [K] value during treatment, (Delta[K]), on the presenting serum [K] concentration,([K]2 ), was: DKA, Delta[K] = 2.78 - 0.81 x [K]2 , r = -0.85, p < 0.001; NKH, Delta[K] = 2.44 - 0.71 x [K]2 , r = -0.90, p < 0.001. The slopes of the regressions were not significantly different. Stepwise logistic regression including both DKA and NKH cases identified the presenting serum [K] level and the change in serum [TCO2 ] value during treatment as the predictors of Delta[K] (R2 = 0.81). Hyperkalemia is a feature of severe hyperglycemia (DKA or NKH) occurring in patients on dialysis. Insulin administration brings about correction of DKA and return of serum [K] concentration to the normal range in the majority of the hyperglycemic episodes without the need for other measures. The initial serum [K] value and the change in serum [TCO2 ] level during treatment influence the decrease in serum [K] value during treatment of dialysis-associated hyperglycemia with insulin. PMID- 15818546 TI - A trial of detecting impending access-graft failure by simplified weekly flow monitoring. AB - Access graft failure is a major problem in hemodialysis. Monitoring the flow through the access so that impending failure can be detected and prevented seems reasonable, but recent clinical trials have failed to show any benefit of such monitoring. Described here are plans for a clinical trial of a new flow monitoring procedure that measures access flow weekly instead of monthly and, being performed before dialysis, avoids the dialysis-induced changes in graft flow that may have affected earlier trials. The planned trial is to be carried out in two stages, the first to establish the sensitivity and specificity of the new method, and the second (if the results of the first stage warrant it) a controlled trial comparing access-costs and hospitalization days between a monitored group and a matched standard care control group. It is hoped that this trial of the new method will establish it as an effective means of extending access-graft life. PMID- 15818547 TI - New fiber configuration for intravenous gas exchange. AB - Implantation of a membrane oxygenator (IO) into the vena cava for blood oxygenation in patients with acute lung failure has been researched for the last 25 years. Compared to the extra corporeal blood oxygenation, where blood is handled outside the body, IO doesn't present tubes, housings or heat exchangers, thus reducing considerably blood contact surface and setting priming volume to zero. Otherwise, restricted space in the vena cava and unadvantageous blood flow conditions represent so far a limitation for sufficient gas exchange. A new fiber configuration for intravenous use is being developed, which increases the implantable fiber surface and enhances gas exchange due to the increased blood convection. This is made possible by new fiber bundles, which are free to slide on a catheter and after implantation assume a twisted shape characterized by high homogeneity and fiber density. PMID- 15818548 TI - Early results of valved bovine jugular vein conduit for right ventricular outflow tract reconstruction. AB - OBJECTIVE: Many congenital cardiac anomalies present with accompanying severe right ventricular outflow tract (RVOT) obstruction or interruption requiring surgical correction. RVOT reconstruction by means of a conduit is often necessary in the majority of these patients in the early years of life and there are several proposed conduits for this purpose. METHODS: Fourteen patients with different congenital cardiac pathologies underwent RVOT reconstruction with the newly developed bovine valved jugular vein conduit (The Contegra conduit). The function of the conduit is observed by echocardiographic examinations at the hospital discharge and at follow-up visits with special attention to the function of the venous valve and to any gradient on the RVOT. RESULTS: There were two perioperative mortalities. All the surviving patients are followed for a mean period of 8.07 months (range 2 to 33 months). The function of the venous valve was determined, in 4 patients (33.33%) as without regurgitation, in 7 patients (58.3%) as mild regurgitation and in 1 patient (8.33%) as mild-to-moderate regurgitation. The reconstructed RVOT was free of any significant gradient at the hospital discharge (mean 10.83 +/-10.18 mmHg) and at the follow-ups (mean 12.916 +/-12.33 mmHg). There was not a trend towards an increase in the gradients following discharge. CONCLUSION: The early results of the Contegra valved conduit are very satisfactory. This graft can be a good alternative for RVOT reconstruction, particularly in the neonatal and infant patient group. These patients can be managed by homograft replacement of their conduit in later years. PMID- 15818549 TI - Design and in vitro performance of a novel bileaflet mechanical heart valve prosthesis. AB - Design and in vitro performance of a novel bileaflet mechanical heart valve prosthesis are presented. The novel heart valve exhibits three main design characteristics: (i) The leaflets form a Venturi passage in open position. Thus, a beneficial pressure distribution is obtained and the leaflets are stabilised in opened position. (ii) The orifice inlet is nozzle-shaped. Flow is convectively accelerated and flow separation at the orifice inlet is avoided. (iii) The hinge design facilitates an additional axial movement of the leaflets and leads to a self cleaning effect and enhances washout of the hinges. The design of the leaflet hinges is of main importance for the functional reliability and durability of mechanical heart valves. After manufacturing first prototypes from titanium and polymeric materials the hydrodynamic performance was evaluated according to ISO 5840 and FDA guidelines. Hydrodynamic performance is comparable with the results of commonly available bileaflet mechanical heart valve prostheses. Initial durability tests showed suitable material couples for further long term studies. PMID- 15818550 TI - Feasibility of continuous CO2 removal with hydrophilic membranes at low blood flow rates. AB - In the conventional treatment of acute respiratory distress syndrome (ARDS), high O2 concentrations and mechanical ventilation may damage the lung tissue. Extracorporeal membrane oxygenation limits damage, provides the needed O2 supply and improves survival of ARDS neonates, but not of adults. Hydrophilic membranes used in hemodialysis are more non-thrombogenic and biocompatible than those used in blood oxygenation, but their O2 transport capacity is not as high. In recent years, CO2 removal at low blood flow rates combined with apneic oxygenation and low frequency ventilation has proved promising in the treatment of ARDS. This approach makes O2 supply across ECMO membranes unnecessary; it also makes hydrophilic membranes candidates for extracorporeal CO2 removal to minimize anticoagulation and immune system activation. This paper reports on the in vitro capacity of hydrophilic polysulphone membranes to remove CO2 from carbonated pig blood into an oxygen-rich gas stream. Experiments were performed on clinical-size dialysis modules and their capacity to remove CO2 as a function of blood flow rate and membrane surface area was investigated. Membranes effectively removed CO2 , more so at increasing blood flow rates and membrane surface areas, at rates of up to 15% of the CO2 metabolic production rate. The specific CO2 removal rate was comparable to that of blood oxygenators equipped with microporous hydrophobic membranes. It is concluded that CO2 removal from slowly flowing blood with hydrophilic membranes is feasible. PMID- 15818551 TI - Phosphorus-enriched hemodialysis for the treatment of patients with severe methanol intoxication. AB - Severe methanol poisoning requires treatment with prolonged and intensive hemodialytic therapy. Such treatment can engender either the de novo development of hypophosphatemia or the worsening of pre-existing hypophosphatemia. Phosphorus enriched hemodialysis therapy can prevent the occurrence of this complication. We report three patients with severe methanol poisoning who were treated with phosphorus-enriched hemodialysis. Prevention or treatment of hypophosphatemia was successfully achieved with this dialytic technique. PMID- 15818552 TI - The dachshund gene is required for the proper guidance and branching of mushroom body axons in Drosophila melanogaster. AB - The dachshund gene encodes a transcription factor required for the proper development of Drosophila eyes, legs, and mushroom bodies. The mushroom bodies of dachshund mutants exhibit a marked reduction in the size of the vertical lobes and disorganization of the horizontal lobes. In mosaic animals, mutant axons fail to contribute significantly to the mushroom body alpha lobe. Here we show that this defect is due to the misrouting of alphabeta axons to the region normally occupied by alpha'beta' axons. This defect is pronounced for clones generated in larval stages but not clones generated after pupariation, indicating that dachshund function is particularly important around the time of puparium formation. In addition, mushroom body axons exhibit excessive branching in dachshund mutant clones. Thus, dachshund is required in mushroom body neurons for proper axon guidance and branching. PMID- 15818554 TI - Synaptic thermoprotection in a desert-dwelling Drosophila species. AB - Synaptic transmission is a critical mechanism for transferring information from the nervous system to the body. Environmental stress, such as extreme temperature, can disrupt synaptic transmission and result in death. Previous work on larval Drosophila has shown that prior heat-shock exposure protects synaptic transmission against failure during subsequent thermal stress. This induced thermoprotection has been ascribed to an up-regulation of the inducible heat shock protein, Hsp70. However, the mechanisms mediating natural thermoprotection in the wild are unknown. We compared synaptic thermosensitivity between D. melanogaster and a desert species, D. arizonae. Synaptic thermosensitivity and the functional limits of the related locomotor behavior differed significantly between closely related, albeit ecologically distinct species. Locomotory behavior of wandering third instar D. arizonae larvae was less thermosensitive and the upper temperature limit of locomotory function exceeded that of D. melanogaster by 6 degrees C. Behavioral results corresponded with significantly lower synaptic thermosensitivity at the neuromuscular junction in D. arizonae. Prior heat-shock protected only D. melanogaster by increasing relative excitatory junctional potential (EJP) duration, the time required for EJP failure at 40 degrees C, and the incidence of EJP recovery following heat-induced failure. Hsp70 induction profiles following heat-shock demonstrate up-regulation of inducible Hsp70 in D. melanogaster but not in D. arizonae. However, expression of Hsp70 under control conditions is greater in D. arizonae. These results suggest that the mechanisms of natural thermoprotection involve an increase in baseline Hsp70 expression. PMID- 15818553 TI - D-Hillarin, a novel W180-domain protein, affects cytokinesis through interaction with the septin family member Pnut. AB - By database searches of the Drosophila genome project we have identified D-hil as the fly member of a novel family of W180-domain containing proteins. Immunocytochemistry demonstrated that D-hil is localized to the neuropil of the embryonic CNS, to the cellular cortex of dividing neuroblasts from larval brains, and that it is up-regulated in the cleavage furrow of S2 cells. We show that D hil distribution overlaps extensively with that of the septin family member Pnut. Cross-immunoprecipitation experiments further indicated that the two proteins may be members of the same protein complex. Analysis of a severe hypomorphic P element mutation in the D-hil locus suggested that D-hil is a nonessential protein. However, by creating double mutant flies we show that the D-hil locus acts as a modulator of Pnut function by increasing the level of polyploidy of neuroblasts in Pnut(KG00478)/Pnut(KG00478) larval brains. Based on these results we propose that D-hil may function as a regulator of septin function during cytokinesis in the developing nervous system. PMID- 15818555 TI - Seasonal-like plasticity of spontaneous firing rate in a songbird pre-motor nucleus. AB - Many animals exhibit seasonal changes in behavior and its underlying neural substrates. In seasonally breeding songbirds, the brain nuclei that control song learning and production undergo substantial structural changes at the onset of each breeding season, in association with changes in song behavior. These changes are largely mediated by photoperiod-dependent changes in circulating concentrations of gonadal steroid hormones. Little is known, however, about whether changes in the electrophysiological activity of neurons accompany the dramatic morphological changes in the song nuclei. Here we induced seasonal-like changes in the song systems of adult white-crowned sparrows and used extracellular recording in acute brain slices from those individuals to study physiological properties of neurons in the robust nucleus of the arcopallium (RA), a pre-motor nucleus necessary for song production. We report that: RA neurons from birds in breeding condition show a more than twofold increase in spontaneous firing rate compared to those from nonbreeding condition; this change appears to require both androgenic and estrogenic actions; and this change is intrinsic to the RA neurons. Thus, neurons in the song circuit exhibit both morphological and physiological adult seasonal plasticity. PMID- 15818556 TI - Estrogen provision by reactive glia decreases apoptosis in the zebra finch (Taeniopygia guttata). AB - Upregulation of aromatase (estrogen synthase) in glia around the site of neural injury may limit neural degeneration. Systemic administration of estrogen limits neural damage, but the specific role of local estrogen provision in this effect is unclear. In male zebra finches, we tested the effect of local aromatase inhibition and estrogen replacement on type of cellular degeneration and the distance of this degeneration from the source of insult. Subjects received injections of the aromatase inhibitor fadrozole into one telencephalic lobe and fadrozole and estradiol into the contralateral lobe. Seventy-two hours later, we used Fluoro-Jade B and TUNEL to label dying and apoptotic cells, respectively. Since each subject was its own control, we were able to assess the influence of local estrogen replacement in relative distinction from circulating steroids and constitutive aromatization. Cellular degeneration around the lesion was measured with Fluoro-Jade B, TUNEL, and indirectly with aromatase expression. Additionally, the glial nature of aromatase-positive cells around the injury was queried by co-localization with vimentin. The estrogen replaced injury had fewer apoptotic cells clustered more closely around the injury compared to the hemisphere injected with fadrozole alone. Since Fluoro-Jade B and TUNEL labeled similar numbers of cells, and the distance of these cells from the injection was identical, we suggest that estrogen replacement functions primarily to restrict apoptosis in the current paradigm. Lastly, aromatase-positive cells around injuries co-localize vimentin, establishing their glial nature. Thus, glial estrogen provision at sites of neural insult may be critical in limiting the cellular degeneration caused by injury via an inhibition of apoptosis. PMID- 15818557 TI - Trifunctional bispecific antibody-induced tumor cell lysis of squamous cell carcinomas of the upper aerodigestive tract. AB - BACKGROUND: The trifunctional bispecific antibody Removab (tbAB) bridges and activates CD3 positive T cells to EpCAM on carcinoma cells and simultaneously binds to an accessory immune-cell inducing tumor cell lysis. tbAB-induced tumor cytotoxicity was assessed in an autologous human ex vivo system. METHODS: One hundred forty tumor samples and autologous peripheral blood mononuclear cells from a total of 36 patients with head and neck squamous cell carcinomas (HNSCCs) were incubated on a chicken embryo chorioallantois membrane with Removab. Tumor cells coincubated with cisplatin or cell culture medium served as positive and negative controls. Tumor cell lysis was assessed by acridine orange staining or by fluorescence-activated cell sorting of propidium iodide-marked cells after 24 and 48 hours (T24/T48) coincubation. RESULTS: Coincubation of HNSCC cells with tbAB and autologous peripheral blood mononuclear cells resulted in a 49% +/- 6% decrease of viable cells at T24 (p < .005) and in a decrease of 56% +/- 8% at T48 (p < .005) compared with the control. The tumor cytotoxicity was similar to that of cisplatin (49% +/- 7% decrease at T24 and 49% +/- 8% at T48). CONCLUSION: In an autologous human ex vivo system, the tbAB-induced tumor cell lysis was comparable to that by cisplatin. PMID- 15818558 TI - Effects of hyperbaric oxygen exposure on experimental head and neck tumor growth, oxygenation, and vasculature. AB - BACKGROUND: Hyperbaric oxygen (HBO2) is used to promote healing in irradiated tissues, but concern persists about the possibility that it may promote residual tumor growth. METHODS: The tumor growth of SQ20B and Detroit 562 head and neck squamous cell carcinoma xenografts were studied after single-dose irradiation and 5x/week HBO2 treatment at 2.4 atm absolute for 90 minutes. The effect of HBO2 treatment on tumor hypoxia and vasculature was also examined by immunohistochemical analysis. RESULTS: HBO2 treatment increased tumor oxygenation during the treatment interval but did not promote the growth of either irradiated or unirradiated tumors. No increase in tumor vascular endothelial growth factor expression or vascularization was detected. CONCLUSIONS: This study found no evidence for persistent changes in tumor microenvironment or tumor growth promotion caused by hyperbaric oxygen exposure. PMID- 15818559 TI - Analysis of bacterial diversity in acidic pond water and compost after treatment of artificial acid mine drainage for metal removal. AB - The microbial population of a sludge amended leaf compost material utilized for treatment of artificial acid mine drainage was studied by culture-independent molecular methods. Iron-rich and sulfurous wastewater (artificial acid mine drainage) was circulated through a column bioreactor for 16 months. After 12 months the column was inoculated with a mixed culture from an acidic pond receiving acid mine drainage from a tailings impoundment at a decommissioned site in Kristineberg, North Sweden. Hydrogen sulfide odor and the formation of black precipitates indicated that sulfate-reduction occurred in the column. 16S rDNA gene analysis by denaturing gradient gel electrophoresis, cloning, and sequencing as well as fluorescent in situ hybridization confirmed the presence of microorganisms closely related to sulfate-reducing bacteria and microorganisms from the genera Pseudoxanthmonas, Dechlorosoma, Desulfovibrio, Agrobacterium, Methylocapsa, Rhodococcus, Sulfobacillus, and some unidentified bacteria. Sulfate reducing bacteria were found in the column bioreactor 2 weeks after inoculation, but not thereafter. This suggests they were in low abundance, even though sulfate remediation rates were significant. Instead, the population contained species similar to those previously found to utilize humic substances released from the compost material. PMID- 15818560 TI - Gene-expression profiles for five key glycosylation genes for galactose-fed CHO cells expressing recombinant IL-4/13 cytokine trap. AB - Recombinant protein glycosylation profiles have been shown to affect the in-vivo half-life, and therefore the efficacy and economics, for many therapeutics. While much research has been conducted correlating the effects of various stimuli on recombinant protein glycosylation characteristics, relatively little work has examined glycosylation-related gene-expression profiles. In this study, the effects of galactose feeding on the gene-expression profiles for five key glycosylation-related genes were determined for Chinese hamster ovary cells producing a recombinant IL-4/13 cytokine trap fusion. The genes investigated were sialidase, a putative alpha2,3-sialyltransferase, CMP-sialic acid transporter, beta1,4-galactosyltransferase, and UDP-galactosyltransferase. Additionally, the sialic acid content (sialylation) of the recombinant protein was examined. The peak sialic acid content of the IL-4/13 cytokine trap fusion protein was observed to be similar for the control and galactose-fed cultures. The gene-expression profiles for four of the glycosylation genes were observed to be sensitive to the glucose concentration and not significantly different for the control and galactose-fed cultures prior to glucose depletion. However, the sialidase gene expression profiles were different for the control and galactose-fed cultures. The sialidase gene-expression profile increased significantly for the galactose fed cultures prior to glucose depletion, whereas for the control cultures, the sialidase gene-expression profiles did not increase until the late stationary phase. The intracellular sialidase enzyme activity decreased exponentially with time for the control cultures; however, for the galactose-fed cultures, the intracellular sialidase enzyme activity decreased initially and then remained relatively high compared to the control cultures. These results indicate that the galactose feeding may increase the potential for desialylation, which offsets any improvements in the sialylation rate due to increased substrate levels. Thus, galactose feeding is an unnecessary expense for the production of the IL-4/13 cytokine trap fusion protein in a batch process. PMID- 15818561 TI - High-efficiency protein expression mediated by enterovirus 71 internal ribosome entry site. AB - An internal ribosome entry site (IRES) has been used to facilitate the expression of more than one protein in a single transcript. In this study, we examined the translational activities of several IRES elements derived from different RNA viruses. The protein expression of encephalomyocarditis virus (EMCV) IRES is similar to that of hepatitis C virus (HCV) IRES in mammalian cells. Notably, the protein expression of enterovirus 71 (EV71) IRES was 23-fold higher than the efficiency of EMCV IRES following normalization of mRNA transcriptional level. Thus, expression of the secreted alkaline phosphatase (SEAP) reporter protein in mammalian cells may be controlled at desirable levels by using appropriate IRES in the expression vector. PMID- 15818562 TI - Stabilization of enzymes by multipoint immobilization of thiolated proteins on new epoxy-thiol supports. AB - The controlled and partial modification of epoxy groups of Eupergit C and EP Sepabeads with sodium sulfide has permitted the preparation of thiol-epoxy supports. Their use allowed not only the specific immobilization of enzymes through their thiol groups via thiol-disulfide interchange, but also enzyme stabilization via multipoint covalent attachment. Penicillin G acylase (PGA) from Escherichia coli and lipase from Rhizomucor miehei were used as model enzymes. Both enzymes lacked exposed cysteine residues, but were introduced via chemical modification under very mild conditions. In the first moments of the immobilization, a certain percentage of immobilized protein could be released from the support by incubation with DTT; this confirms that the first step was via a thiol-disulfide interchange. Moreover, the promotion of some further epoxy enzyme bonds was confirmed because no enzyme release was detected after some immobilization time by incubation with DTT. In the case of the heterodimeric PGA, it was possible to demonstrate the formation of at least one epoxy bond per enzyme subunit by analyzing with SDS-PAGE the supernatants obtained after boiling the enzyme derivatives in the presence of mercaptoethanol and SDS. Thermal inactivation studies showed that these multipoint enzyme-support attachments promoted an increase in the stability of the immobilized enzymes. In both cases, the stabilization factor was around 12-15-fold comparing optimal derivatives with their just-thiol immobilized counterparts. PMID- 15818563 TI - Production of recombinant plant gum with tobacco cell culture in bioreactor and gum characterization. AB - Many plant gums, such as gum arabic, contain hydroxyproline-rich glycoproteins (HRGPs), which are also abundant components of the plant cell extracellular matrix. Here we expressed in transgenic BY2 Nicotiana tabacum (tobacco) cells, a synthetic gene encoding a novel HRGP-based gum, designated gum arabic-8 or (GA)(8). (GA)(8) encoded eight repeats of the consensus polypeptide sequence of gum arabic glycoprotein (GAGP): Gly-Pro-His-Ser-Pro-Pro-Pro-Pro-Leu-Ser-Pro-Ser Pro-Thr-Pro-Thr-Pro-Pro-Leu, in which most of the Pro residues were posttranslationally modified to hydroxyproline (Hyp). (GA)(8) was expressed as a green fluorescent protein (GFP) fusion protein targeted to the culture medium, (GA)(8)GFP. The culture of the transgenic cells in a 5-L bioreactor showed that the production of (GA)(8)GFP was cell growth-associated. The extracellular yield of (GA)(8)GFP was 116.8 mg/L after 14 days of culture and accounted for 87% of the total fusion protein expressed. (GA)(8)GFP was purified from the culture medium by a combination of hydrophobic interaction, gel permeation, and reversed phase chromatography. Biochemical characterization indicated that the amino acid composition of the (GA)(8) module, after removal of GFP by proteolysis, was virtually identical to that of predicted by the GAGP consensus sequence and that carbohydrate, which occurred as arabinogalactan polysaccharides and small oligoarabinosides O-linked through the Hyp residues, accounted for 84% of the molecules' dry weight. Functional assays showed that (GA)(8) exhibited low viscosity in aqueous solution similar to native GAGP. However, neither GFP alone nor the (GA)(8) module could emulsify orange oil. However, the fusion protein (GA)(8)GFP possessed 1.28-fold better emulsification properties than native GAGP. This work demonstrates the feasibility and potential of a synthetic gene approach to the de novo design of novel glycoprotein-based gums and emulsifiers. PMID- 15818564 TI - Mechanistic studies on aggregation of polyethylenimine-DNA complexes and its prevention. AB - Aggregation of polyethylenimine (PEI)-DNA complexes severely undermines their utility for gene delivery into mammalian cells. Herein we undertook to elucidate the mechanism of this deleterious phenomenon and to develop rational strategies for its prevention. The effect of temperature, surfactants, complex concentration, ionic strength, viscosity, and pH on the time course of this aggregation was systematically examined. The aggregation process was completely inhibited by 2.5% polyoxyethylene (100) stearate (POES) and to a lesser degree by other nonionic surfactants. Importantly, POES preserved the transfection efficiency of the complexes without inducing toxicity. The aggregation was also reduced by lowering the temperature and pH, diluting the complexes, and increasing the solution viscosity. It is concluded that PEI-DNA complexes aggregate primarily due to hydrophobic interactions, while electrostatic attractions play little role. PMID- 15818565 TI - Reduction of Cr(VI) by immobilized cells of Desulfovibrio vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776. AB - Hexavalent chromium, a carcinogen and mutagen, can be reduced to Cr(III) by Desulfovibrio vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776. This study examined Cr(VI) reduction by immobilized cells of the two strains in a common solution matrix using various entrapment matrices. Chitosan and PVA-borate beads did not retain integrity and supported low or no reduction of Cr(VI) by the cells. A commercial preparation (Lentikats) was stable but also did not support Cr(VI) reduction. K-carrageenan beads were stable in batch suspensions but gel integrity was lost after only 5 h in a flow-through system in the presence of 100 microM Cr(VI). The best immobilization matrices were agar and agarose, where the initial rates of reduction of Cr(VI) (from 500 microM solution) for D. vulgaris NCIMB 8303 and Microbacterium sp. NCIMB 13776 were 127 (agar) and 130 (agarose), and 15 (agar) and 12 (agarose) nmol h(-1) mg dry cell wt(-1), respectively. The higher removal of Cr(VI) by D. vulgaris was also seen in 14-mL packed-bed flow through columns, where, at a flow rate of 2.4 mL h(-1), the percentage removal of Cr(VI) was approximately 95% and 60% for D. vulgaris and Microbacterium sp., respectively (agar-immobilized cells). The Cr(VI) reducing activities of D. vulgaris and Microbacterium sp. were lost after 159 and 140 h, respectively. Examination of the beads for structural integrity within the columns in situ using magnetic resonance imaging after 24 and 100 h of continuous operation against Cr(VI) (with negligible Cr retained within the columns) showed that agar beads were more stable with time. The most appropriate system for development of a continuous bioprocess is thus the use of D. vulgaris NCIMB 8303 immobilized in an agar gel matrix. PMID- 15818566 TI - Synthesis and structure-activity relationship of 4-substituted benzoic acids and their inhibitory effect on the biosynthesis of fatty acids and sterols. AB - The synthesis of 4-[3-(substituted phenyl)-2-oxo-5-oxazolidinyl]methoxybenzoic acids and their inhibitory effects on the biosynthesis of fatty acids and sterols is described. IC50 values in vitro were 10(-6) and 10(-5) M, respectively. Though the in vitro inhibitory activity of all these compounds toward sterol biosynthesis was inferior to that of pravastatin, several compounds had a stronger reducing effect in Sprague-Dawley (SD) rat on both, cholesterol (TC) and triglyceride (TG), than pravastatin and bezafibrate. The potent compounds were present at high concentrations in rat liver. The enantiomers of the potent racemic compounds (4-[3-(4-bromo-2-fluorophenyl)-2-oxo-5 oxazolidinyl]methoxybenzoic acid) were prepared and their activity was examined in vivo and in vitro. In vivo, each enantiomer possessed more activity than the racemic compound. Further, in Watanabe hereditable hyperlipidemic (WHHL) rabbit, optically active (R)-4-[3-(4-bromo-2-fluorophenyl)-2-oxo-5 oxazolidinyl]methoxybenzoic acid also potently reduced the effect of both TC and TG on serum levels, compared with pravastatin and bezafibrate. PMID- 15818567 TI - Using the diphosphanyl radical as a potential spin label: effect of motion on the EPR spectrum of an R1(R2)P--PR1 radical. AB - The EPR spectrum of the novel radical Mes*(CH3)P--PMes* (Mes*=2,4,6-(tBu)3C6H2) was measured in the temperature range 100-300 K, and was found to be drastically temperature dependent as a result of the large anisotropy of the 31P hyperfine tensors. Below 180 K, a spectrum of the liquid solution is accurately simulated by calculating the spectral modifications due to slow tumbling of the radical. To achieve this simulation, an algorithm was developed by extending the well-known nitroxide slow-motion simulation technique for the coupling of one electron spin to two nuclear spins. An additional dynamic process responsible for the observed line broadening was found to occur between 180 K and room temperature; this broadening is consistent with an exchange between two conformations. The differences between the isotropic 31P couplings associated with the two conformers are shown to be probably due to an internal rotation about the P--P bond. PMID- 15818568 TI - C-linked disaccharide analogue of the Thomsen-Friedenreich (T)-epitope alpha-O conjugated to L-serine. AB - Condensation of a silylated beta-D-galactopyranosylaldehyde (3) with isolevoglucosenone (4) in the presence of Et(2)AlI provided bicyclic enone 5. Subsequent addition of BnNHOMe gave adduct 6, which was converted into 4-O-acetyl 1,6-anhydro-3-C-[(1 R)-1,3,4,5,7-penta-O-acetyl-2,6-anhydro-D-glycero-L-manno heptitol-1-C-yl]-2-azido-2,3-dideoxy-beta-D-galacto-hexopyranose after liberation of the 2-amino group, its transformation into a 2-azido moiety, desilylation, and peracetylation. Ring-opening of the 1,6-anhydro galactopyranosyl unit and O glycosidation with Fmoc-Ser-O-tBu afforded a 5:1 mixture of alpha- and beta galactosides. Treatment with CH(3)COSH gave pure N-[(9H-fluoren-9 ylmethoxy)carbonyl]-{4,6-di-O-acetyl-3-C-[(1 R)-2,6-anhydro 1,3,4,5,7-penta-O acetyl-D-glycero-L-manno-heptitol-1-C-yl]-2-[(N-acetyl)amino]-2,3-dideoxy-alpha-D galactopyranosyl}-l-serine tert-butyl ester (2), a protected form of a C disaccharide analogue of the Thomsen-Friedenreich (or T) epitope (beta-D-Galp-(1- >3)-alpha-D-GalNAcp) alpha-O-conjugated to L-serine. PMID- 15818569 TI - Electronic structural changes between nickel(II)-semiquinonato and nickel(III) catecholato states driven by chemical and physical perturbation. AB - The selective synthesis of tetracoordinate square-planar low-spin nickel(II) semiquinonato (Ni(II)-SQ) and nickel(III)-catecholato (Ni(III)-Cat) complexes, 1 and 2, respectively, was achieved by using bidentate ligands with modulated nitrogen-donor ability to the nickel ion. The electronic structures of 1 and 2 were revealed by XPS and EPR measurements. The absorption spectra of 1 and 2 in a noncoordinating solvent, dichloromethane (CH2Cl2), are completely different from those in tetrahydrofuran (THF), being a coordinating solvent. As expected from this result, the gradual addition of N,N-dimethylformamide (DMF), which is also a coordinating solvent like THF, into a solution of 1 or 2 in CH2Cl2 leads to color changes from blue (for 1) and brown (for 2) to light green, which is the same color observed for solutions of 1 or 2 in THF. Furthermore, the same color changes are induced by varying the temperature. Such spectral changes are attributable to the transformation from square-planar low-spin Ni(II)-SQ and Ni(III)-Cat complexes to octahedral high-spin Ni(II)-SQ ones, caused by the coordination of two solvent molecules to the nickel ion. PMID- 15818570 TI - NMR spectral assignments of three aspidofractinine alkaloids, kopsine, fruticosine and fruticosamine. AB - 1D and 2D NMR techniques were used to assign fully the spectra of three aspidofractinine alkaloids, kopsine (1), fruticosamine (2) and fruticosine (3), isolated from a cultivated specimen of the plant Kopsia fruticosa (Apocynaceae). The assignment of the NMR data for 1, 2 and 3 will help in the future assignments of related alkaloids. PMID- 15818571 TI - A prospective study of ultrasound screening for molar pregnancies in missed miscarriages. AB - OBJECTIVE: To examine the relationship between ultrasound and histological features in the screening for molar changes in missed miscarriage. METHODS: A prospective cohort study was conducted on all missed miscarriages, with features suspicious of molar pregnancy, on transvaginal ultrasound and/or on histological examination over a 5-year period. All cases of molar pregnancy diagnosed histologically were examined and cross-referenced with cases diagnosed on ultrasound and with the supplementary report from the regional referral center. When available, maternal serum beta-human chorionic gonadotropin (hCG) levels were recorded. RESULTS: Fifty-one cases of suspected molar pregnancy were referred to the regional center for further histological opinion and follow-up, and five cases were subsequently excluded from the final analysis because of the diagnosis of hydropic abortion (HA). In 33 cases a molar pregnancy was suspected at the initial scan. Of these, 26 (78.8%) were confirmed on histology, resulting in a 56% detection rate using ultrasound alone. In 15 cases hCG results were available, of which nine were greater than two multiples of the median. CONCLUSIONS: The diagnosis of both complete (CHM) and partial (PHM) hydatidiform moles in first-trimester miscarriages is difficult. hCG is significantly higher in both CHM and PHM and, in conjunction with transvaginal ultrasound, could provide the screening test required to enable clinicians to counsel women more confidently towards non-surgical methods of management of their miscarriage, where histopathological examination is not available. PMID- 15818572 TI - Down-regulation of a morphogen (sonic hedgehog) gradient in the gastric epithelium of Helicobacter pylori-infected Mongolian gerbils. AB - Sonic hedgehog (Shh) is a morphogen involved in many aspects of patterning of the gut during embryogenesis and in gastric fundic gland homeostasis in the adult. Intestinal metaplastic change of the gastric epithelium is associated with the loss of Shh expression, and mice that lack Shh expression show intestinal transformation of the gastric mucosa. The present study was designed to investigate the alteration of Shh expression in the stomach of an experimental model of Helicobacter pylori (H. pylori) colonization. Male Mongolian gerbils were inoculated with H. pylori and examined 4 and 51 weeks later. The level of Shh mRNA expression was determined by quantitative RT-PCR and in situ hybridization. Shh protein expression was determined by immunoblotting and immunohistochemistry. Shh was expressed in the parietal cells, zymogenic cells, and mucous neck cells of the gastric fundic glands of gerbils. Prolonged colonization by H. pylori led to extension of the inflammation from the antrum to the corpus of the stomach, with loss of Shh expression. Loss of Shh expression correlated with loss of parietal cells, disturbed maturation of the mucous neck cell-zymogenic cell lineage, and increased cellular proliferation. Shh expression is significantly reduced in H. pylori-associated gastritis. These data show for the first time that H. pylori infection leads to down-regulation of the expression of a morphogen with an established role (Shh) in gastric epithelial differentiation. PMID- 15818573 TI - Homozygosity for a CHEK2*1100delC mutation identified in familial colorectal cancer does not lead to a severe clinical phenotype. AB - It has recently been suggested that the frequency of the germline CHEK2*1100delC mutation is higher among breast cancer families with colorectal cancer, although the mutation does not seem to be significantly associated with familial colorectal cancer. Five hundred and sixty-four familial colorectal tumours were studied for expression of CHEK2 using tissue microarrays and an antibody against the NH2-terminal SQ regulatory domain of the CHEK2 protein. Normal colonic tissue from patients whose tumours showed loss of CHEK2 expression was investigated further using fragment and sequence analysis for the presence of a CHEK2*1100delC mutation and five other (R117G, R137Q, R145W, I157T, and R180H) known germline variants in CHEK2. Twenty-nine tumours demonstrated loss of expression for CHEK2. Analysis of matched normal colonic tissue from these patients revealed germline CHEK2*1100delC mutation in three cases. In two of these, the mutation was heterozygous but, interestingly, the third patient proved to be homozygous for the deletion, using six different primer pair combinations. None of the other tested germline variants were identified. No CHEK2*1100delC mutations were found in patients whose tumours stained positive. Homozygosity for the CHEK2*1100delC mutation appears not to be lethal in humans. No severe clinical phenotype was apparent, although the patient died from colonic carcinoma at age 52 years. This observation is in line with recent knockout mouse models, although in the latter, cellular defects in apoptosis and increased resistance to irradiation seem to exist. It is also concluded that CHEK2 protein abrogation is not caused by the CHEK2 germline variants R117G, R137Q, R145W, I157T, and R180H in familial colorectal cancer. PMID- 15818574 TI - Analytical potential of 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein for the determination of amino compounds by capillary electrophoresis with laser-induced fluorescence detection. AB - The analytical potential of a fluorescein analogue, 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein (SAMF), for the first time synthesized in our laboratory, as a labeling reagent for the labeling and determination of amino compounds by capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection was investigated. Biogenic monoamines and amino acids were chosen as model analytes to evaluate the analytical possibilities of this approach. The derivatization conditions and separation parameters for the biogenic amines were optimized in detail. The derivatization was performed at 30 degrees C for 6 min in boric acid buffer (pH 8.0). The derivatives were baseline-separated in 15 min with 25 mM boric acid running buffer (pH 9.0), containing 24 mM SDS and 12.5% v/v acetonitrile. The concentration detection limit for biogenic amines reaches 8 x 10(-11) mol.L(-1) (signal-to-noise ratio = 3). The application of CE in the analysis of the SAMF derivatized amino acids was also exploited. The optimal running buffer for amino acids suggested that weak acidic background electrolyte offered better separation than the basic one. The proposed method was applied to the determination of biogenic amines in three different beer samples with satisfying recoveries varying from 92.8% to 104.8%. Finally, comparison of several fluorescein-based probes for amino compounds was discussed. With good labeling reaction, excellent photostability, pH-independent fluorescence (pH 4-9), and the resultant widely suited running buffer pH, SAMF has a great prospect in the determination of amino compounds in CE. PMID- 15818575 TI - Trace analysis of oxidized, nitrated, and chlorinated aromatic amino acids by capillary electrophoresis with electroosmotic flow modification allowing large volume sample stacking. AB - A capillary electrophoresis method has been developed for the simultaneous analysis of the oxidized, nitrated, and chlorinated aromatic amino acids, as well as their parent compounds. These modifications of the aromatic amino acids in proteins or free form are induced by the attack of reactive, mainly free radical species generated during cell stress, and these stable products may serve as biomarkers of cell damage. The analytes tyrosine, phenylalanine, dihydroxyphenylalanine, tryptophan, 3-nitrotyrosine, 3-chlorotyrosine, ortho tyrosine, meta-tyrosine, 3-hydroxyphenylacetic acid (internal standard 1), and alpha-methyltyrosine (internal standard 2) were separated in their anionic forms in alkaline borate buffer. The polyamine spermine was used as electroosmotic flow (EOF) modifier. Adsorbing to the capillary wall, spermine can either suppress or even reverse the EOF depending on its concentration and the pH. The effects of the pH of the separation buffer, the spermine concentration, the temperature, and the applied field strength on the separation were examined. The modified aromatic amino acids are present in biological fluids in a much lower concentration than their parent compounds, thus high detection sensitivity of the analytical method is required. To achieve good detection sensitivity, field-amplified sample stacking of large injection volumes was applied. Omitting polyamine from the sample buffer allowed local reversal of the EOF, thus removal of the low conductivity sample buffer at the capillary inlet. In this way, 100% of the capillary to the detection window could be filled with the sample, and the detection limits achieved for the modified aromatic amino acids were in the range of 2.5-10 nM. PMID- 15818576 TI - Synthesis and characterization of quaternary ammonium dicarboxylic acid isoelectric buffers and their use in pH-biased isoelectric trapping separations. AB - Two approaches are described in this paper for the synthesis of isoelectric buffers that have pI values in the 1.5 < pI < 4.3 range. The first synthesis relies on the alkylation of existing aminodicarboxylic acids and recovery of the ampholyte as an inner salt. The second synthesis method forms low-pI ampholytes by reacting a secondary amine with two equivalents of an alkylester of a haloalkanecarboxylic acid, followed by hydrolysis of the intermediate in an alkaline solution and recovery of the ampholyte as an inner salt. The new ampholytes have been analytically characterized by capillary electrophoresis, high-resolution electrospray ionization-mass spectrometry, one- and two dimensional nuclear magnetic resonance (NMR) spectroscopy, and X-ray crystallography. The isoionic solutions of the new ampholytes have high buffering capacity and conductivity, making them good pH biasers in the receiving stream in preparative-scale pH-biased isoelectric trapping separations. PMID- 15818577 TI - Conductivity detection in capillary zone electrophoresis: inspection by PeakMaster. AB - A simple rule stating that the signal in conductivity detection in capillary zone electrophoresis is proportional to the difference between the analyte mobility and mobility of the background electrolyte (BGE) co-ion is valid only for systems with fully ionized electrolytes. In zone electrophoresis systems with weak electrolytes both conductivity signal and electromigration dispersion of analyte peaks depend on the conductivity and pH effects. This allows optimization of the composition of BGEs to give a good conductivity signal of analytes while still keeping electromigration dispersion near zero, regardless of the injected amount of sample. The demands to achieve minimum electromigration dispersion and high sensitivity in conductivity detection can be accomplished at the same time. PeakMaster software is used for inspection of BGEs commonly used for separation of sugars (carbohydrates, saccharides) at highly alkaline pH. It is shown that the terms direct and indirect conductivity detection are misleading and should not be used. PMID- 15818578 TI - Synthesis of heptakis(2-O-methyl-3-O-acetyl-6-O-sulfo)-cyclomaltoheptaose, a single-isomer, sulfated beta-cyclodextrin carrying nonidentical substituents at all the C2, C3, and C6 positions and its use for the capillary electrophoretic separation of enantiomers in acidic aqueous and methanolic background electrolytes. AB - The sodium salt of heptakis(2-O-methyl-3-O-acetyl-6-O-sulfo)cyclomaltoheptaose (HMAS), the first single-isomer, sulfated beta-cyclodextrin carrying nonidentical substituents at all of the C2, C3, and C6 positions, has been synthesized, analytically characterized, and used for the capillary electrophoretic separation of the enantiomers of a group of 24 weak base pharmaceuticals in acidic aqueous and acidic methanolic background electrolytes. HMAS interacted more strongly with most of the analytes studied than heptakis(2,3-di-O-methyl-6-O sulfo)cyclomaltoheptaose, but less strongly than heptakis(2,3-di-O-acetyl-6-O sulfo)cyclomaltoheptaose, the respective analogs with identical substituents at the C2 and C3 positions. The good separation selectivities and favorable normalized electroosmotic flow mobilities allowed for rapid, efficient separation of the enantiomers of 19 of the 24 weak base analytes in the aqueous and methanolic background electrolytes. The trends in the effective mobilities and separation selectivities as a function of the HMAS concentration closely followed the predictions of the ionic strength-corrected charged resolving agent migration model. PMID- 15818579 TI - Thermosensitive biodegradable polydepsipeptide. AB - A poly(N-isopropylacrylamide) (PNIPAAm)-like biodegradable thermosensitive polydepsipeptide, poly[Glc-Asn(N-isopropyl)], was synthesized by introducing an isopropyl amide group into poly[Glc-Asn]. Poly[Glc-Asn(N-isopropyl)] was degraded in vitro by cleavage of the ester bonds in the main chain in water at room temperature. The non-toxic nature of the polymer and its degradation products, coupled with a cloud point at 29 degrees C in water, make this polymer attractive for biomedical implant applications. PMID- 15818580 TI - Synthesis and chemical recycling of novel poly(ester-urethane)s using an enzyme. AB - A series of enzymatically recyclable poly(ester-urethane)s consisting of a biodegradable diurethane moiety as a hard segment and an ester moiety as an enzymatically cleavable linkage was chemo-enzymatically prepared by two routes. The poly(ester-urethane) was prepared by a) the ring-opening polymerization of a cyclic ester-urethane monomer synthesized via the transesterification reaction of biodegradable diurethanediol and dicarboxylate ester using lipase and b) the direct polycondensation of a diurethanediol and a dicarboxylate ester. A significantly higher molecular-weight poly(ester-urethane) having the highest molecular weight (Mw) of 101,000 was produced by the ring-opening polymerization of the cyclic ester-urethane monomer when compared with that produced by the polycondensation of the dicarboxylate ester with diurethanediol. The poly(ester urethane) was readily degraded by lipase into the corresponding cyclic oligomers; the oligomers were readily repolymerized by the ring-opening polymerization using lipase for chemical recycling. PMID- 15818581 TI - Controlled release of goserelin from microporous polyglycolide and polylactide. AB - Two microporous biodegradable polyesters, i.e., PGA and PDLLA, were obtained by solid-state polymerization reaction from the sodium salts of the corresponding alpha-hydroxycarboxylic acids after washing out the by-product sodium chloride. The polymers were shaped by cold uniaxial pressing, by hot uniaxial pressing, and by extrusion at elevated temperature. Due to the special microporosity of the polymers, the introduction of drugs is possible at moderate temperature. The release kinetics of the model drug Phe and of the anti-tumor drug goserelin (an LH-RH agonist) from compacted polymer samples were fast (approx. 2 d). The release kinetics of goserelin were corrected for the decomposition of the drug. External coatings with PDLLA or PLLA obtained by immersion in polymer solution strongly slowed down the release kinetics in the case of the PDLLA coating, giving an almost linear release during 100 d. A coating with PLLA was unsuitable to slow down the release kinetics. PMID- 15818582 TI - Polymeric micelles and nanoparticles from block and statistical poly((RS)-3,3 dimethylmalic acid) derivatives: preparation and characterization. AB - Amphiphilic and biodegradable micelles and nanoparticles designed as potential drug carriers were prepared from biodegradable statistical and block copolyesters obtained by a living anionic ring-opening process. These novel materials display amphiphilic properties arising from the joint presence of hydrophilic poly((RS) 3,3-dimethylmalic acid) and hydrophobic poly(hexyl (RS)-3,3-dimethylmalate) segments. Micelles obtained from a well-defined block copolymer have been characterized by their critical aggregation concentration, and nanoparticles derived from statistical copolymer have been analyzed by transmission electron microscopy (TEM). PMID- 15818583 TI - Associative pH sensitive system. AB - New associative pH sensitive systems composed of a modified dextran bearing 2 carboxycyclohexyl carboxyl groups and neutral (p betaCD) or positively charged (p betaCDN(+))beta-cyclodextrin-co-epichlorhydrin copolymers have been synthesized and their properties were investigated as a function of pH by phase diagrams and viscosimetry. The affinity between the CD cavities and the hydrophobic guests (2 carboxycyclohexyl carboxyl groups) has been studied as a function of pH. The system with the neutral p betaCD shows a pH dependent behavior due to the lowest affinity of the guest at pH above 5 for the cavity. Associative phase separation is produced at low pH whereas soluble complexes occur at higher pH. The opposite trend is observed with the system containing the cationic p betaCDN(+). This behavior is due to the combination of electrostatic and inclusion complex interactions in this system. Addition of salt, by screening the electrostatic interactions, also strongly influences the response of the system. PMID- 15818584 TI - Structure of water incorporated in sulfobetaine polymer films as studied by ATR FTIR. AB - The structure and hydrogen bonding of water in the vicinity of a thin film of a sulfobetaine copolymer (poly[(N,N-dimethyl-N-(3-sulfopropyl)-3' methacrylamidopropanaminium inner salt)-ran-(butyl methacrylate)], poly(SPB-r BMA)), were analyzed with band shapes of O-H stretching of attenuated total reflection infrared (ATR-IR) spectra. The copolymer could be cast as a thin film, of approximate thickness 10 microm, on a ZnSe crystal for the ATR-IR spectroscopy. At an early stage of sorption of water into the polymer film, the O H stretching band of the IR spectra for the water incorporated in the film was similar to that for free water. This is consistent with the tendency for another zwitterionic polymeric material, poly[(2-methacryloyloxyethylphosphorylcholine) ran-(butyl methacrylate)] (poly(MPC-r-BMA). It is, however, contradictory to the drastic change in the O-H stretching band for water incorporated into films of polymers such as poly(2-hydroxyethyl methacrylate), poly(methyl methacrylate) and poly(butyl methacrylate). These results suggest that polymers with a zwitterionic structure do not significantly disturb the hydrogen bonding between water molecules incorporated in the thin films. The investigation into the blood compatibility of both the poly(SPB-r-BMA) and the poly(MPC-r-BMA) films indicate a definite correlation between the blood-compatibility of the polymers and the lack of effect of the polymeric materials on the structure of the incorporated water. PMID- 15818585 TI - Novel acrylonitrile-based copolymers containing phospholipid moieties: synthesis and characterization. AB - Novel acrylonitrile-based copolymers containing phospholipid moieties were synthesized by a three-step process, which included the copolymerization of acrylonitrile and 2-hydroxyethyl methacrylate (HEMA) in water and the reaction of the resulting poly[acrylonitrile-co-(2-hydroxyethyl methacrylate)]s (PANCHEMA) with 2-chloro-2-oxo-1,3,2-dioxaphospholane (COP) followed by the ring-opening reaction of COP with trimethylamine. The chemical structure of PANCHEMA and the phospholipid-containing acrylonitrile-based copolymers (PLCANCP) was analyzed with FT-IR spectroscopy, (1)H and (31)P NMR, and XPS. Surface properties of the studied copolymers were evaluated by the pure water contact angle, protein adsorption and platelets adhesion measurements. The water contact angle measured by sessile drop method decreased for the polymers in the following sequence: PAN, PANCHEMA, and PLCANCP. The adsorption amount of bovine serum albumin and the adhesive number of platelet followed the same decline sequence. These results demonstrate that the biocompatibility of the acrylonitrile-based copolymer membranes could be improved efficiently by the introduction of phospholipid moieties. PMID- 15818586 TI - Small-angle X-ray scattering on solutions of carboxymethylcellulose and bovine serum albumin. AB - The existence of attractive interaction between CMC and BSA was evidenced in solution at pH higher than the protein isoelectric point by means of SAXS. Mixtures of BSA at 10 x 10(-3) g . mL(-1) and CMC at the concentration range of 1 x 10(-3) g . mL(-1) to 10 x 10(-3) g . mL(-1) were investigated. Upturns in the very low q range revealed the presence of aggregates when the CMC concentration was higher than 2 x 10(-3) g . mL(-1). The comparison between the calculated form factor with the experimental curves at intermediate and long q values indicated that the BSA molecules keep their native form in mixtures with CMC concentrations lower than 5 x 10(-3) g . mL(-1). Therefore, for CMC concentrations higher than 2 x 10(-3) g . mL(-1) the mixtures start to present aggregates and free BSA molecules coexisting in solution. PMID- 15818588 TI - Electrochemical addressing of the optical properties of a monolayer on a transparent conducting substrate. PMID- 15818587 TI - The properties of covalently immobilized trypsin on soap-free P(MMA-EA-AA) latex particles. AB - The covalent immobilization of trypsin onto poly[(methyl methacrylate)-co-(ethyl acrylate)-co-(acrylic acid)] latex particles, produced by a soap-free emulsion polymerization technique, was carried out using the carbodiimide method. The catalytic properties and kinetic parameters, as well as the stability of the immobilized enzyme were compared to those of the free enzyme. Results showed that the optimum temperature and pH for the immobilized trypsin in the hydrolysis of casein were 55 degrees C and 8.5, both of which were higher than that of the free form. It was found that K(m) (Michaelis constant) was 45.7 mg . ml(-1) and V(max) (maximal reaction rate) was 793.0 microg . min(-1) for immobilized trypsin, compared to a K(m) of 30.0 mg . ml(-1) and a V(max) of 5 467.5 microg . min(-1) for free trypsin. The immobilized trypsin exhibited much better thermal and chemical stabilities than its free counterpart and maintained over 63% of its initial activity after reusing ten times. PMID- 15818589 TI - Ga10Br10(4-tert-Butylpyridine)10: a mixed-valent gallium(I) subhalide as an intermediate during the formation of elemental gallium? PMID- 15818590 TI - From racemic primary aminoalkyl(phosphanyl)ferrocene complexes to a lithium phosphorus closo cluster. PMID- 15818592 TI - Good communication with patients receiving bad news about cancer in Japan. AB - PURPOSE: Despite a recent increase in the attention given to improving communication when disclosing bad news to cancer patients, understanding of good patient-physician communication is still lacking in Japan. The aim of the present study is to describe attributes of good communication with patients receiving bad news about cancer in Japan. METHODS: Forty-nine participants, including 42 cancer patients and seven oncologists, were requested to undergo an in-depth interview, and the data obtained were qualitatively analyzed. RESULTS: Seventy-one attributes of good communication with patients receiving bad news about cancer were obtained from 619 opinions, and classified into four domains. (1) SETTING: a private room and sufficient time. (2) Information given: impact on daily activities and alternative therapy. (3) Method of disclosure: facilitation of patient understanding. (4) Emotional support: encouragement and allowing expression of emotions. CONCLUSIONS: The four domains of good patient-physician communication in Japan are apparently similar to those in Western countries. The domain involving information given and method of disclosure in previous Western studies was further divided into two domains in the present study. Japanese physicians should perhaps pay careful attention to these issues when disclosing bad news about cancer. There are some cultural differences related to the details of communication attributes (e.g. using euphemisms and showing consideration for the patient's family). PMID- 15818594 TI - Evidence that prostate gonadotropin-releasing hormone receptors mediate an anti tumourigenic response to analogue therapy in hormone refractory prostate cancer. AB - Gonadotropin-releasing hormone analogue (GnRHa) therapy is an established method of androgen withdrawal in the treatment of prostate cancer. The present study investigated if the expression of prostate GnRH receptors (GnRHRs) might influence the response to GnRHa. GnRHR protein expression was first studied in a panel of prostate cancer cell lines. In androgen-dependent cells, GnRHR expression was unchanged following acute or chronic androgen withdrawal. In these cells, GnRHa significantly inhibited androgen-induced cell proliferation (p = 0.01). In contrast, GnRHa was unable to further suppress basal levels of cell proliferation induced by androgen withdrawal. In androgen-independent prostate cancer cells, variable levels of GnRHR expression were observed. In these cells, GnRHa treatment blocked cell proliferation (p = 0.001) and invasion (up to 70%) induced by fibroblast growth factor stimulation. Crucially, this effect was only evident in cells that expressed high levels of the GnRHR. GnRHa treatment also significantly inhibited the ability of these cells to recover from a cytotoxic insult (50% inhibition). The clinical significance of prostate GnRHR was tested by immunohistochemistry in a preliminary cohort of patients treated with GnRHa or surgical castration. There was no association between GnRHR expression and pathological grade, clinical stage, time to PSA nadir (p = 0.82) (n = 35) or progression to hormone refractory disease (p = 0.22) (n = 21), irrespective of the treatment method. GnRHa therapy in the presence of high GnRHR expression however, was found to be associated with longer disease-specific survival (mean survival 85 months, p = 0.002). In contrast, high GnRHR expression was not associated with survival among surgically castrated patients (mean survival 50 months, p = 0.7). Taken together, these data support the notion of a functional interaction between GnRHa and the GnRHR, which results in an anti-tumourigenic effect on prostate cancer cells. Findings from this report have direct implications for the use of GnRHR as a novel therapeutic target in hormone refractory prostate cancer. PMID- 15818593 TI - Diagnostic value of OCT3/4 for pre-invasive and invasive testicular germ cell tumours. AB - Human testicular germ cell tumours of adolescents and adults (TGCTs), the seminomatous and non-seminomatous germ cell tumours, show morphological and biological similarities to normal embryonic development, presumably determined by their supposed cell of origin, the primordial germ cell/gonocyte. Based on this knowledge, OCT3/4, also known as POU5F1, was recently defined as a diagnostic marker for these tumour types. In the adult testis, positive immunohistochemistry for OCT3/4 is an absolute indicator for the presence of the TGCT precursor carcinoma in situ/intratubular germ cell neoplasia undifferentiated (CIS/ITGCNU), seminoma, and/or embryonal carcinoma. Several studies have confirmed this observation, using the same polyclonal antibody. The present study demonstrates the usefulness of OCT3/4 immunohistochemistry in a diagnostic setting of a consecutively collected series of more than 200 testicular tumours and over 80 testicular biopsies. Moreover, it is shown that a monoclonal antibody directed against OCT3/4 is as informative as the polyclonal antibody, both in immunohistochemistry and in western blot analysis. The antibodies are robust and applicable with different methods of pretreatment and storage of tissue. This allows routine application of this diagnostic marker. PMID- 15818595 TI - Electron ionization fragmentation mechanisms of different substituted 2,3-dihydro and 2,3,4,5-tetrahydro-1,5-benzothiazepines. AB - The electron ionization induced fragmentations of ten biologically significant 2,3-dihydro-1,5-benzothiazepines and the corresponding 2,3,4,5-tetrahydro-1,5 benzothiazepines have been studied by low- and high-resolution mass spectrometry. The fragmentations follow a general pattern, the details of which are discussed with respect to the nature and position of the substituent in the aromatic ring. The dihydro- and tetrahydro-1,5-benzothiazepines both undergo fragmentation through four routes (A-D). However, the most significant fragmentation takes place through route A, leading to the elimination of ring A or ring B of the molecule. The difference between the fragmentation patterns of dihydro- and tetrahydro-1,5-benzothiazepines appears mainly in route E where a phenylallylhydroxybenzene cation appears in all tetrahydro-1,5-benzothiazepines but is not observed in the corresponding dihydro derivatives. PMID- 15818596 TI - Families of long-term cancer survivors: health maintenance advocacy and practice. AB - Research on cancer and aging has addressed health maintenance issues for older adults in long-term cancer survivorship, but not their family members. The current study focused on two aspects of health maintenance for families: advocacy and practice. Regression analyses explored the effects of demographic characteristics and perceptions during diagnoses and treatments on family members' health maintenance in long-term survivorship. Family members were acting as health maintenance advocates for their surviving relatives. Advocacy was related to being a caregiver during diagnosis and treatment. In terms of their own health, family members were practicing a high number of health maintenance activities. Practice was associated with being a caregiver during diagnosis and treatment and with being older and White. Implications for interventions and future research with family members are discussed. PMID- 15818597 TI - Impaired capacity for upregulation of MHC class II in tumor-associated microglia. AB - Immunotherapy for malignant gliomas is being studied as a possible adjunctive therapy for this highly fatal disease. Thus far, inadequate understanding of brain tumor immunology has hindered the design of such therapies. For instance, the role of microglia and macrophages, which comprise a significant proportion of tumor-infiltrating inflammatory cells, in the regulation of the local anti-tumor immune response is poorly understood. To study the response of microglia and macrophages to known activators in brain tumors, we injected CpG oligodeoxynucleotide (ODN), interferon-gamma (IFN-gamma), and IFN-gamma/LPS into normal and intracranial RG2 glioma-bearing rodents. Microglia/macrophage infiltration and their surface expression of MHC class II B7.1 and B7.2 was examined by flow cytometry. Each agent evaluated yielded a distinct microglia/macrophage response: CpG ODN was the most potent inducer of microglia/macrophage infiltration and B7.1 expression, while IFN-gamma resulted in the highest MHC-II expression in both normal and tumors. Regardless of the agent injected, however, MHC-II induction was significantly muted in tumor microglia/macrophage as compared with normal brain. These data suggest that microglia/macrophage responsiveness to activators can vary in brain tumors when compared with normal brain. Understanding the mechanism of these differences may be critical in the development of novel immunotherapies for malignant glioma. PMID- 15818598 TI - Transactivation of cyclin E gene by EWS-Fli1 and antitumor effects of cyclin dependent kinase inhibitor on Ewing's family tumor cells. AB - Chromosomal translocation t(11; 22)(q24; q12) is detected in approximately 90% of Ewing's family tumors (EFTs) including Ewing's sarcoma and primitive neuroectodermal tumor. This results in the formation of the EWS-Fli1 fusion gene, which produces EWS-Fli1 fusion protein. This chimerical gene product acts as an aberrant transcriptional activator, which may be responsible for the tumorigenesis of EFTs. We have previously reported that cyclin E expression was upregulated in EFT cells and in EWS-Fli1 transformed fibroblastic cells. However, the mechanism of the overexpression of cyclin E by EWS-Fli1 is still unknown. In our study, we investigated the mechanism of transactivation of the cyclin E gene in EFT cells. We found that EWS-Fli1 enhanced the activity of the cyclin E gene promoter partially through E2F binding sites in the promoter. In addition, the basic transcriptional factor, Sp1, might also be involved in the transactivation of the cyclin E gene by EWS-Fli1. To study the biological significance of cyclin E overexpression in EFT cells, we used flavopiridol, a pan-cyclin-dependent kinase (CDK) inhibitor and found that flavopiridol efficiently suppressed the growth of EFT cells in vitro and in vivo by the inhibition of cyclinE/CDK2 kinase activity and the induction of apoptosis. These results suggest that targeting of the cyclin/CDK complex may provide new insight into treatment of EFTs. PMID- 15818599 TI - Evidence-based medical review update: pharmacological and surgical treatments of Parkinson's disease: 2001 to 2004. AB - The objective of this study is to update a previous evidence-based medicine (EBM) review on Parkinson's disease (PD) treatments, adding January 2001 to January 2004 information. The Movement Disorder Society (MDS) Task Force prepared an EBM review of PD treatments covering data up to January 2001. The authors reviewed Level I (randomized clinical trials) reports of pharmacological and surgical interventions for PD, published as full articles in English (January 2001-January 2004). Inclusion criteria and ranking followed the original program and adhered to EBM methodology. For Efficacy Conclusions, treatments were designated Efficacious, Likely Efficacious, Non-Efficacious, or Insufficient Data. Four clinical indications were considered for each intervention: prevention of disease progression; treatment of Parkinsonism, as monotherapy and as adjuncts to levodopa where indicated; prevention of motor complications; treatment of motor complications. Twenty-seven new studies qualified for efficacy review, and others covered new safety issues. Apomorphine, piribedil, unilateral pallidotomy, and subthalamic nucleus stimulation moved upward in efficacy ratings. Rasagiline, was newly rated as Efficacious monotherapy for control of Parkinsonism. New Level I data moved human fetal nigral transplants, as performed to date, from Insufficient Data to Non- efficacious for the treatment of Parkinsonism, motor fluctuations, and dyskinesias. Selegiline was reassigned as Non-efficacious for the prevention of dyskinesias. Other designations did not change. In a field as active in clinical trials as PD, frequent updating of therapy-based reviews is essential. We consider a 3-year period a reasonable time frame for published updates and are working to establish a Web-based mechanism to update the report in an ongoing manner. PMID- 15818600 TI - Integrating the criminal justice system into mental health service delivery: the worcester diversion experience. AB - The substantial number of persons with mental illness encountered in many sectors of the criminal justice system has spurred actors from various agencies within that system to take actions aimed at reducing the growth of this population. These actions have included the development of specialty police units, jail diversion programs, and other mechanisms for channeling persons with mental illness out of the criminal justice system and into mental health treatment. The courts, too, have become involved in this effort with the recent development of the "mental health court," the latest of the "specialty" or "problem solving courts." These courts have not been without their critics, however, nor are they the only feasible approach to court-based diversion. This paper identifies and explores a range of options for structuring the relationship between criminal courts and local mental health systems. Beginning with a discussion of the rationale motivating the development of mental health courts, two alternatives to this specialty court model are discussed. One involves judges dealing with defendants having mental illness and substance abuse on a case-by-case basis. The other takes advantages of linkages that may already exist between most courts and the mental health providers who conduct their forensic assessments, expanding the role of these providers to serve as boundary spanners between courts and the components of local mental health systems. Regardless of the model adopted, however, appropriate linkages must exist between the courts and relevant providers. A case study is provided that demonstrates how the status of a locale's linkages can be evaluated and how the information derived from such evaluation can be used to improve the linkages between police, courts, and health and human services agencies. PMID- 15818601 TI - XMR, a dual location protein in the XY pair and in its associated nucleolus in mouse spermatocytes. AB - Xlr and Xmr are sex-specific genes which are expressed during the meiotic prophase I in the mouse. In spermatocytes, XMR concentrates on the asynapsed regions of the XY chromosomes, suggesting that XMR plays a role in sex chromosome condensation and silencing. The present study shows that in the mouse, XMR also concentrates in the nucleolus which is closely associated with the XY chromosome pair. In this species, the formation of a large fibrillo-granular nucleolus signals the activation of the ribosomal genes, but release of pre-ribosomal particles is inhibited. Using laser confocal microscopy we characterized the distribution of XMR in the XY body relative to the XY chromatin and the nucleolus. Immunoelectron microscopy showed that XMR concentrates in the fibrillo granular component and the granular component (GC) of the nucleolus. In (T[X;16]16H) mouse spermatocytes, the nucleolus displays little or no activity and does not associate with the XY pair. XMR concentrated only on the XY chromosomes in (T[X;16]16H) mouse spermatocytes. These data suggest that XMR could play a role both in the XY pair and the nucleolus associated to the sex chromosomes. PMID- 15818602 TI - Introduction to this issue: diversion from the criminal justice system. PMID- 15818603 TI - Evaluating the efficiency and community safety goals of the Broward County Mental Health Court. AB - Mental health courts have developed as one response to persons with mental illness who are involved with the criminal justice system. This study investigated the efficiency and safety goals of one such court in Broward County, FL. Mental health court (MHC) clients spent significantly fewer days in jail for the index arrest associated with study enrollment than a comparison group. MHC clients had similar survival time to re-arrest up to one year after study enrollment. MHC clients did not significantly differ from the comparison group in self-reported aggressive acts over an 8 month follow-up period, while they did self-report significantly fewer acts of violence than the comparison group at the 8 month follow-up. These findings suggest that some of the benefits associated with the MHC reported in prior studies were not achieved at the expense of efficiency and safety. PMID- 15818604 TI - From referral to disposition: case processing in seven mental health courts. AB - The number of mental health courts in the United States is rapidly increasing, from one in 1997 to nearly 100 in 2004. However, to date there is comparatively little research regarding these specialty courts. The present study reports data on the referral and disposition decision-making processes of seven mental health courts. Information on all referrals to the seven courts over a three-month period was gathered. Results show that, in comparison with individuals involved in the criminal justice system, mental health court clients are more likely to be older, White, and women than individuals in the general criminal justice system. Furthermore, this over-representation occurs at the point of referral, rather than at the point of the court's decision to accept or reject a referral. In addition, the length of time from referral to diversion is much longer in these mental health courts than in other types of diversion programs. Implications of these findings are discussed. PMID- 15818605 TI - Accuracy of self-reported arrests among a forensic SPMI population. AB - Self-reports are prone to response error even in the most structured and standardized environment, but the highly stigmatizing nature of illegal behavior and the possible cognitive problems inherent to the SPMI population increase the likelihood of such error. Using administrative data as the gold standard, the validity of self-reported arrests was examined among 85 mentally ill substance abusing individuals with justice involvement. The overall accuracy of self reported arrests during a three-month recall period was 84.7%. Validity was higher among individuals who at the time of the follow-up interview were either in a jail diversion program or incarcerated. Individuals with a mood disorder appeared to be somewhat less accurate in their self-reports than subjects with schizophrenia, despite the implication of cognitive distortion accompanying this latter diagnostic category. Further research on issues such as the accuracy of self-reports over longer recall periods, or the accuracy of self-reported frequency of arrest, is necessary, given the reliance on self-report methodology in outcome studies. PMID- 15818606 TI - Victim-centric diversion? The electronic monitoring of domestic violence cases. AB - This article discusses the electronic monitoring of domestic violence cases as a form of diversion. The data derive from a two and a half year study of two "bilateral" electronic monitoring programs geared towards domestic violence offenders, mostly defendants, and their former or current intimate partners. Analysis consists of an examination of how the programs under study manifest the aims and tensions of diversion, and highlights how a victim-centered approach to diversion can amplify the tensions of diversion. PMID- 15818607 TI - Assessing the effectiveness of jail diversion programs for persons with serious mental illness and co-occurring substance use disorders. AB - In recent years, jail diversion programs for people with serious mental illness and co-occurring substance use disorders have received increasing attention and have rapidly grown in number. Previous studies suggest that jail diversion programs have the potential to achieve positive outcomes. The present study reports findings from 6 jail diversion programs (3 pre-booking and 3 post booking) participating in a federally-funded research initiative to assess the effectiveness of jail diversion programs for people with co-occurring disorders. Diverted and non-diverted groups were compared on self-reported outcomes at 12 months following diversion. The findings suggest that jail diversion reduces time spent in jail without increasing the public safety risk, while linking participants to community-based services. Jail diversion costs and the implications of these results for jail diversion programs and future research are discussed. PMID- 15818608 TI - Contrasting jail diversion and in-jail services for mental illness and substance abuse: do they serve the same clients? AB - Baseline data from a study of jail diversion services and in-jail behavioral health services were used to examine the differences in clients served by these two models of responding to people with co-occurring mental health and substance abuse problems in the criminal justice system. Clients of the diversion service had more acute psychiatric symptoms and were more likely to have a diagnosis of psychosis NOS. Clients of the in-jail service were more likely to have been on probation or parole in the past and to have received substance abuse treatment. Different service models may attract and serve different populations of clients. Diversion services may cast a wider net that includes clients who may not have otherwise been involved in forensic services. PMID- 15818609 TI - Efficacy of a Mental Health Treatment Court with assertive community treatment. AB - This study examined the efficacy of a Mental Health Treatment Court (MHTC) with diversion to treatment supported by an assertive community treatment (ACT) model of case management. A total of 235 participants were randomly assigned to either MHTC or treatment as usual (TAU) and assessed over a 2 year period. It was hypothesized that participants in the MHTC would decrease their criminal activity and improve their psychosocial functioning relative to participants receiving TAU. While there were offenders for whom neither treatment was effective, a majority in both groups decreased jail days and improved psychosocial functioning, with MHTC participants demonstrating greater gains in most areas. The impact of implementing the MHTC on community practices, and the value of integrating criminal justice and mental health systems, is discussed. PMID- 15818610 TI - A cluster randomized controlled trial of visual, cytology and human papillomavirus screening for cancer of the cervix in rural India. AB - The impact of screening by visual inspection with acetic acid (VIA), cytology or HPV testing on cervical cancer incidence and mortality is investigated in a cluster randomized controlled trial in India. We report findings after the screening phase, when 52 clusters, with a total of 142,701 women aged 30-59 years in Osmanabad District, India, were randomized into 4 arms for a single round of screening by trained midwives with either VIA, cytology or HPV testing as well as a control group. All laboratory tests were done locally. Test-positive women underwent investigations (colposcopy/biopsy) and treatment in the base hospital. Data on participation, test positivity, positive predictive value and detection rates of cervical neoplasia were analyzed using cluster design methodology. Of the eligible women, 72-74% were screened. Test positivity rates were 14.0% for VIA, 7.0% for cytology and 10.3% for HPV. The detection rate of high-grade lesions was similar in all intervention arms (0.7% for VIA, 1.0% for cytology and 0.9% for HPV testing) (p = 0.06, Mann-Whitney test). While the detection rate for VIA dropped to 0.5% with declining test positivity during the course of the study, it remained constant for cytology and HPV testing. Over 85% of women with high-grade lesions received treatment. Our results show that a high level of participation and good-quality cytology can be achieved in low-resource settings. VIA is a useful alternative but requires careful monitoring. Detection rates obtained by HPV testing were similar to cytology, despite higher investments. PMID- 15818611 TI - Perceived deterrence and outcomes in drug court. AB - According to perceived-deterrence theory, the likelihood that an offender will engage in drug use or illegal activity is influenced by the perceived certainty of being detected for infractions or recognized for accomplishments, the perceived certainty of receiving sanctions for infractions or rewards for accomplishments, and the anticipated magnitude of the sanctions and rewards. This study evaluated drug court participants' perceived deterrence at monthly intervals during their enrollment in drug court. Exploratory cluster analysis (N=255) on the longitudinal scores yielded five subtypes of drug offenders characterized either by consistently elevated perceived-deterrence scores, consistently moderate scores, consistently low scores, increasing scores, or decreasing scores. The best outcomes were associated with consistently elevated scores, whereas the worst outcomes were associated with scores that declined over time as the participants became accustomed to the program. The clusters also differed in predicted directions on demographic variables. The correlational design does not permit inferences of causality; however, the results lend credence to perceived deterrence as a potential explanatory mechanism for the effects of drug courts. PMID- 15818612 TI - Peroxisome proliferator-activated receptor gamma agonist troglitazone induces colon tumors in normal C57BL/6J mice and enhances colonic carcinogenesis in Apc1638 N/+ Mlh1+/- double mutant mice. AB - The role of the nuclear peroxisome proliferator-activated receptor-gamma (PPAR gamma) in colon tumorigenesis remains controversial. Notwithstanding evidence that PPAR-gamma ligands impede murine colorectal carcinogenesis, PPAR-gamma agonists have been shown to enhance in vivo tumor formation in mouse models of human colon cancer. Our study was designed to determine whether troglitazone (TGZ) induces colonic tumor formation in normal C57BL/6J mice and enhances colorectal carcinogenesis in double mutant Apc1638N/+ Mlh1+/- mice fed a standard AIN-76A diet. We report herein that not only does TGZ enhance carcinogenesis in the large intestine of mutant mice predisposed to intestinal carcinogenesis but TGZ also induces colonic tumors in normal mice without gene targeting or carcinogen administration. This observation indicates that preexisting mutational events are not necessary for induction of colonic tumors by activated PPAR-gamma in vivo. PMID- 15818613 TI - DNA damage uncouples the mitogenic response to IGF-I in MCF-7 malignant breast cancer cells by switching the roles of PI3 kinase and p21WAF1/Cip1. AB - In addition to its mitogenic and survival actions, recent evidence indicates that IGF-I can enhance DNA repair, implying IGF activity may limit the efficacy of many therapeutic strategies that rely on induction of DNA damage. Although the individual pathways by which DNA damage and IGF-I act are well understood, the cross-talk between these signaling events is not well defined. We examined the effects of DNA damage on the IGF-I response of MCF-7 breast cancer cells. Cells were exposed to the UV mimetic, 4-NQO, or the gamma-irradiation mimetic and chemotherapeutic drug, bleomycin; cellular proliferation was assessed by cell counting, tritiated thymidine incorporation and FACS cell cycle analysis. Although IGF-I acutely suppressed the p53 response to both agents, it subsequently enhanced the chronic increase in p53 and p21(WAF1/Cip1), resulting in cell cycle arrest; however, no apoptosis was observed. Use of specific inhibitors demonstrated that PI3 kinase was utilized with p38 MAPK to induce the p53 response to DNA damage, but was also utilized by IGF-I to diminish the acute p53 response. In addition, p21WAF1/Cip1 was increased by IGF-I, which has previously been shown to contribute to the mitogenic response. Here we demonstrate that with DNA damage IGF-I can also enhance the chronic p53-dependent increase in p21WAF1/Cip1, culminating in growth arrest. Overall, we have shown that PI3 kinase and p21WAF1/Cip1 play dual roles in mediating the mitogenic response to IGF-I, but these are both switched by cellular DNA damage to mediate a growth arrest. PMID- 15818614 TI - Intracellular anti-E7 human antibodies in single-chain format inhibit proliferation of HPV16-positive cervical carcinoma cells. AB - The E7 tumor antigen of human papillomavirus type 16 (HPV16) is a validated target for immunodiagnosis and immunotherapy of HPV-associated cervical cancer. Anti-HPV16 E7 antibodies in scFv format were isolated from a human antibody phage display library and characterized. With the aim of interfering with the oncogenic activity of E7 protein, the most reactive of the selected antibody fragments was expressed by eukaryotic vectors in different compartments of the HPV16-positive cervical carcinoma SiHa cell line. The intracellular antibodies (intrabodies) were tested for their ability of inhibiting cell proliferation. A significant inhibition was obtained targeting the intrabodies to the nuclear and secretory compartments whereas no significant effect was observed in case of cytoplasmic localization. Inhibition was highly specific as no antiproliferative effect was obtained either with the E7-specific intrabodies in HPV-negative cells nor with irrelevant intrabodies in SiHa cells. PMID- 15818615 TI - Family history as a co-factor for adenocarcinoma and squamous cell carcinoma of the uterine cervix: results from two studies conducted in Costa Rica and the United States. AB - Previous work suggests that cervical cancer may aggregate in families. We evaluated the association between a family history of gynecological tumors and risk of squamous cell and adenocarcinomas of the cervix in 2 studies conducted in Costa Rica and the United States. The Costa Rican study consisted of 2,073 women (85 diagnosed with CIN3 or cancer, 55 diagnosed with CIN2 and 1,933 controls) selected from a population-based study of 10,049 women. The U.S. study consisted of 570 women (124 with in situ or invasive adenocarcinomas, 139 with in situ or invasive squamous cell carcinomas of the cervix and 307 community-based controls) recruited as part of a multicentric case-control study in the eastern part of the United States. Information on family history of cervical and other cancers among first-degree relatives was ascertained via questionnaire. Information on other risk factors for cervical cancer was obtained via questionnaire. Human papillomavirus (HPV) exposure was assessed in both studies using broad spectrum HPV L1-based PCR testing of exfoliated cervicovaginal cells and in Costa Rica by additional testing of plasma collected from participants for antibodies against the L1 protein of HPV types 16, 18, 31 and 45 by ELISA. A family history of cervical cancer in a first-degree relative was associated with increased risk of squamous tumors in both studies (odds ration [OR] = 3.2 for CIN3/cancer vs. controls; 95% confidence interval [CI] = 1.1-9.4 in Costa Rica; OR = 2.6 for in situ/invasive squamous cell carcinoma cases vs. controls, 95% CI = 1.1-6.4 in the Eastern United States study). These associations were evident regardless of whether the affected relative was a mother, sister or daughter of the study participant. Furthermore, observed effects were not strongly modified by age. In Costa Rica, the effect persisted in analysis restricted to HPV-exposed individuals (OR = 3.0; 95% CI = 1.0-9.0), whereas in the Eastern United States study there was evidence of attenuation of risk in analysis of squamous carcinoma cases restricted to HPV positive women (OR = 1.4; 95% CI = 0.29-6.6). No significant association was observed between a family history of cervical cancer in a first-degree relative and adenocarcinomas (OR = 1.3; 95% CI = 0.43-3.9). History of gynecological tumors other than cervical cancer in a first-degree relative was not significantly associated with risk of disease in either study. These results are consistent with a role of host factors in the pathogenesis of squamous cell cervical cancer, although familial aggregation due to shared environmental exposures cannot be ruled out. PMID- 15818616 TI - hTERT gene copy number is not associated with hTERT RNA expression or telomerase activity in colorectal cancer. AB - In a majority of malignant human tumors telomerase activity can be detected, suggesting an immortal phenotype. Expression of the reverse transcriptase subunit, hTERT, in the human telomerase complex is required for telomerase activity. The regulation of hTERT, from gene level to a fully functional protein, is still a poorly understood process. Increased copy number of the hTERT gene has been demonstrated in a significant portion of established cell lines and tumors of different origin but its relevance for telomerase activity levels is unclear. In the present study, we examined the hTERT gene copy number using fluorescence in situ hybridization (FISH) in samples from 64 colorectal carcinomas and an increased copy number (> or = 3 hTERT gene copies/nucleus) was observed in 31 cases (48%). No statistical association existed between hTERT gene copy number and hTERT RNA expression or telomerase activity. However, a significant relationship was found between an increase in hTERT gene copy number and p53 protein accumulation (p = 0.002) and aneuploidy (p = 0.036). Only 4 tumors showed microsatellite instability, 3 of which had a normal hTERT gene copy number. The data indicated that the increased copy number of the hTERT gene in colorectal carcinoma was a result of genomic instability with no obvious consequence for telomerase activity levels. PMID- 15818617 TI - Loss of Cyclin B1 followed by downregulation of Cyclin A/Cdk2, apoptosis and antiproliferation in Hela cell line. AB - Recent studies have shown that Cyclin B1 is overexpressed in various tumor types but present at low levels in normal tissues. To explore the possibility of employing Cyclin B1 as an anticancer target, we knocked down Cyclin B1 in an HeLa cell line using RNA interference (RNAi). Subsequently, we monitored cell cycle related molecules by Western blot together with immunofluorescence and determined cell cycle distribution by flow cytometry. XTT and soft agar colony growth experiments were performed to detect cell viability and proliferation. Furthermore, we analyzed cell apoptosis by measuring Bcl-2 and Bax protein level and DNA-ladder assay. After performing Cyclin B1 RNAi, Cyclin B1, Cyclin A and Cdk2 protein levels were found to be markedly downregulated, whereas Cdc2 was almost unaffected; S-phase fraction increased significantly; HeLa cell viability and cell colony forming ability were markedly diminished after the RNAi; Bcl-2 was noticeably attenuated but Bax was hardly changed; and HeLa cells displayed typical DNA ladder. The loss of Cyclin B1 resulted in the downregulation of Cyclin A and Cdk2, S-phase delay and eventually led to cell apoptosis and the decrease of cell viability and proliferation. Our studies suggest that Cyclin B1 may be a promising anticancer target. PMID- 15818618 TI - High-throughput protein expression analysis using tissue microarray technology of a large well-characterised series identifies biologically distinct classes of breast cancer confirming recent cDNA expression analyses. AB - Recent studies on gene molecular profiling using cDNA microarray in a relatively small series of breast cancer have identified biologically distinct groups with apparent clinical and prognostic relevance. The validation of such new taxonomies should be confirmed on larger series of cases prior to acceptance in clinical practice. The development of tissue microarray (TMA) technology provides methodology for high-throughput concomitant analyses of multiple proteins on large numbers of archival tumour samples. In our study, we have used immunohistochemistry techniques applied to TMA preparations of 1,076 cases of invasive breast cancer to study the combined protein expression profiles of a large panel of well-characterized commercially available biomarkers related to epithelial cell lineage, differentiation, hormone and growth factor receptors and gene products known to be altered in some forms of breast cancer. Using hierarchical clustering methodology, 5 groups with distinct patterns of protein expression were identified. A sixth group of only 4 cases was also identified but deemed too small for further detailed assessment. Further analysis of these clusters was performed using multiple layer perceptron (MLP)-artificial neural network (ANN) with a back propagation algorithm to identify key biomarkers driving the membership of each group. We have identified 2 large groups by their expression of luminal epithelial cell phenotypic characteristics, hormone receptors positivity, absence of basal epithelial phenotype characteristics and lack of c-erbB-2 protein overexpression. Two additional groups were characterized by high c-erbB-2 positivity and negative or weak hormone receptors expression but showed differences in MUC1 and E-cadherin expression. The final group was characterized by strong basal epithelial characteristics, p53 positivity, absent hormone receptors and weak to low luminal epithelial cytokeratin expression. In addition, we have identified significant differences between clusters identified in this series with respect to established prognostic factors including tumour grade, size and histologic tumour type as well as differences in patient outcomes. The different protein expression profiles identified in our study confirm the biologic heterogeneity of breast cancer and demonstrate the clinical relevance of classification in this manner. These observations could form the basis of revision of existing traditional classification systems for breast cancer. PMID- 15818619 TI - Ribonucleotide reductase inhibitors enhance cidofovir-induced apoptosis in EBV positive nasopharyngeal carcinoma xenografts. AB - In nasopharyngeal carcinoma (NPC), Epstein-Barr virus (EBV) infection is mainly latent, and the tumor cells contain episomal viral DNA. We have shown that the acyclic nucleoside phosphonate analog, cidofovir [(S)-1-(3-hydroxy-2 (phosphonylmethoxypropyl))cytosine] (HPMPC), inhibits growth of NPC xenografts in nude mice by causing apoptosis. The ribonucleotide reductase (RR) inhibitors, hydroxyurea and didox (3,4-dihydroxybenzohydroxamic acid), have been demonstrated to inhibit neoplastic growth and are used as antiviral and anticancer agents. Here we show that RR inhibitors enhance the antitumor effect of cidofovir in EBV transformed epithelial cells. MTT assays indicate that hydroxyurea and didox enhance cidofovir-induced cell toxicity in NPC-KT cells, an EBV-positive epithelial cell line derived from NPC. The effect is due to enhancement of apoptosis through the caspase cascade as shown by pronounced cleavage of poly(ADP ribose) polymerase. Finally, hydroxyurea strikingly enhanced the cidofovir induced growth-inhibitory effect on NPC grown in athymic mice. The results suggest that RR inhibitors should enhance the antitumor effect of acyclic nucleoside phosphonate analogs on NPC. PMID- 15818620 TI - Identification of 20 genes aberrantly methylated in human breast cancers. AB - Aberrant methylation of CpG islands (CGI) not only plays a role in gene silencing, but is also a potential cancer biomarker. To identify more CGI aberrantly methylated in human breast cancers, we carried out a genome-wide search for aberrant methylation, using methylation-sensitive-representational difference analysis. CGI in 5' upstream regions of 20 genes, TSPAN-2, AK5, LOC284999, HOXD11, FLJ25161, XT3, PCDH10, PCDHGB6, SIM1, LOC346978, COE2, TDH (FLJ25033), LOC346419, FLJ33790, GJB2, AMN, LOC201164, DLX4, DCC and FOXA2, were found to be methylated in at least one of 8 breast cancer cell lines. Fifteen of the 20 genes were methylated in more than one of 21 primary breast cancers in Stages I or II, and especially, those of LOC346978, HOXD11, SIM1, PCDHGB6 and FLJ25161 were methylated in more than 10 cancers. All the breast cancers had some aberrant methylation. Among the 13 genes whose CGI were completely methylated in one or more cell lines, FOXA2 and XT3 were expressed in normal human mammary epithelial cells (HMEC) and were not expressed in cancer cell lines with complete methylation. The other 11 genes examined were barely expressed, or not expressed even in HMEC. Our results showed that breast cancer cells accumulate aberrant methylation of the CGI identified here. This may serve as markers for early-stage breast cancers and suggests that aberrant methylation targets transcriptionally inactive genes in vivo. PMID- 15818621 TI - Endogenous immune response to gangliosides in patients with confined prostate cancer. AB - Our study investigated whether endogenous IgM antibodies to gangliosides occur in patients with early stages of prostate cancer (CaP) patients, after defining ganglioside profiles of CaP cell lines. Immune and resorcinol staining detected the presence of gangliosides GM3, GM2, GD3, GD2 and GD1a but not GM1a, GD1b or GT1b in the extracts of normal prostatic epithelial cells (PrEC) and neoplastic androgen-insensitive (PC-3, DU145) and -sensitive (LNCaP-FGC and LNCaP-FGC-10) CaP cells. Using a sensitive ELISA, developed and validated in our laboratory, the titers of IgM against 8 gangliosides from sera of patients with benign prostatic hyperplasia (BPH) (n = 11), organ-confined (T1/T2, n = 36) and unconfined (T3/T4, n = 27) CaP and age-matched healthy men (n = 11) were determined double-blinded. Using ANOVA and Fisher's least significant difference (LSD) methods, the log-titers among different groups were compared. CaP patients differed from healthy and BPH patients in increased titers against GD1a and decreased titers against GD3. Titers of antibodies to other gangliosides exhibited no difference between CaP patients and others. The specific augmentation of anti-GD1a IgM in patients with organ-confined CaP (stage T1/T2) but not in patients with unconfined CaP (stage T3/T4) or BPH or in healthy controls is striking. This finding together with identification of GD1a as a major ganglioside in CaP cell lines and with the accruing studies on the immunosuppressive nature of GD1a indicates that augmentation of anti-GD1a IgM in confined CaP may signify an early endogenous immune response to eliminate a "danger signal" from tumor microenvironment and circulation. PMID- 15818622 TI - Novel platinum(IV) complexes induce rapid tumor cell death in vitro. AB - The anticancer activity of platinum complexes has been known since the discovery of classical Pt(II)-based drug cisplatin. However, Pt(IV) complexes have greater inertness than corresponding Pt(II) complexes, thus allowing the oral administration and reducing the toxicity associated with platinum-based chemotherapy. Here, we describe the in vitro antitumor activity of some novel Pt(IV)-based agents against mouse fibrosarcoma L929 cells and human astrocytoma U251 cells. The cytotoxicity of 2 Pt(IV) complexes with bidentate ethylenediamine N,N'-di-3-propanoato esters was found to be markedly higher than that of their Pt(II) counterparts and comparable to the antitumor action of cisplatin. In contrast to cisplatin, which caused oxidative stress-independent apoptotic cell death of tumor cells, these Pt(IV) complexes induced oxygen radical-mediated tumor cell necrosis. Importantly, the cytotoxic action of novel Pt(IV) complexes was markedly more rapid than that of cisplatin, indicating their potential usefulness in anticancer therapy. PMID- 15818623 TI - Risk factors for thyroid cancer: a prospective cohort study. AB - Given the higher incidence rate of thyroid cancer among women compared to men and evidence that smoking and alcohol consumption may be inversely related to thyroid cancer risk, we examined thyroid cancer risk in association with menstrual, reproductive and hormonal factors, and cigarette and alcohol consumption, in a prospective cohort study of 89,835 Canadian women aged 40-59 at recruitment who were enrolled in the National Breast Screening Study (NBSS). Linkages to national cancer and mortality databases yielded data on cancer incidence and deaths from all causes, respectively, with follow-up ending between 1998 and 2000. Cox proportional hazards models (using age as the time scale) were used to estimate hazard ratios and 95% confidence intervals for the association between each of the potential risk factors and risk of thyroid cancer overall and by the main histologic subtypes. During a mean of 15.9 years of follow-up, we observed 169 incident thyroid cancer cases. There was no evidence of altered overall thyroid cancer risk with any of the menstrual, reproductive, or hormonal factors. There was evidence of a decreased risk of papillary thyroid cancer among women with 5 or more live births (vs. nulliparous). Age at which smoking commenced, duration of smoking, number of cigarettes smoked per day, pack-years of smoking and alcohol consumption were not associated with altered thyroid cancer risk. The present study provides little support for associations with hormonal factors, smoking, or alcohol consumption, but there is a need for additional prospective data. PMID- 15818624 TI - Serum matrix metalloproteinase-9 in head and neck squamous cell carcinoma is a prognostic marker. AB - The aim of this study was to determine whether serum matrix metalloproteinase-9 (MMP-9) could predict cause-specific and relapse-free survival in patients with squamous cell carcinoma of head and neck. Furthermore, this study was designed to investigate whether there is an association between MMP-9 immunohistochemical staining and serum MMP-9 levels. Pretreatment serum levels of MMP-9 were quantitatively measured by ELISA assay in 67 patients presenting with a primary head and neck squamous cell carcinoma. The results were compared with the corresponding immunohistochemical staining results, clinical data and the patients' outcome. The follow-up time for all of the patients was at least 5 years. There was a statistically significant correlation between circulating MMP 9 and MMP-9 immunohistochemical staining in the corresponding tumors (p = 0.028). The cause-specific and relapse-free survival rates were clearly lower among patients with high MMP-9 serum levels (> 73 ng/ml). The 5-year cause-specific survival-rate was 40% in a patient group with high serum MMP-9, and 69% for patients with a low MMP-9 level (p = 0.027). In the same follow-up period, the cumulative relapse-free survival rate was 36% in patients presenting with a high serum MMP-9 and 66% in those with a low MMP-9 level. No correlation was found between MMP-9 serum levels and the traditional clinical or histopathologic factors. The results suggest for the first time that pretreatment serum MMP-9 level could serve as a prognostic factor in head and neck squamous cell carcinoma. PMID- 15818625 TI - Epidemiology of testicular cancer: an overview. AB - Testicular cancer is a rare disease, accounting for 1.1% of all malignant neoplasms in Canadian males. Despite the low overall incidence of testicular cancer, it is the most common malignancy among young men. The incidence rate of testicular cancer has been increasing since the middle of the 20th century in many western countries. However, the etiology of testicular cancer is not well understood. A search of the peer-reviewed literature was conducted to identify important articles for review and inclusion in this overview of the epidemiology of testicular cancer. Most of the established risk factors are related to early life events, including cryptorchidism, carcinoma in situ and in utero exposure to estrogens. Occupational, lifestyle, socioeconomic and other risk factors have demonstrated mixed associations with testicular cancer. Although there are few established risk factors for testicular cancer, some appear to be related to hormonal balance at various life stages. Lifestyle and occupational exposures occurring later in life may play a role in promoting the disease, although they are not likely involved in cancer initiation. In addition to summarizing the current epidemiologic evidence on risk factors for testicular cancer, we suggest future research directions that may elucidate the etiology of testicular cancer. PMID- 15818626 TI - 2,4-Dinitrobenzenesulfonyl fluoresceins as fluorescent alternatives to Ellman's reagent in thiol-quantification enzyme assays. PMID- 15818627 TI - Corroboration of a familial chordoma locus on chromosome 7q and evidence of genetic heterogeneity using single nucleotide polymorphisms (SNPs). AB - Chordoma, a rare bone tumor originating from notochordal remnants, has a genetic predisposition in some families. Previously, we performed linkage analysis using microsatellite (STR) markers on 3 unrelated chordoma kindreds (16 patients with chordoma) and reported significant evidence for linkage to chromosome 7q33 (Z(max) = 4.78) with a minimal disease gene region of 11 cM. In our present study, we performed linkage analysis in these 3 families using chromosome 7 single nucleotide polymorphisms (SNPs). Parametric and nonparametric multipoint analyses showed significant linkage to 7q markers with p < 0.001 and Z(max) = 2.77, respectively. The minimal disease gene region was not reduced by combined SNP and STR haplotype analysis compared to the previous STR haplotype analysis alone. We genotyped members of a fourth chordoma family with SNP and STR markers for chromosome 7q and for 1p36, the location of a previously reported chordoma locus. Affected members of this family did not share a common haplotype on 7q, and the family did not show evidence of linkage to 1p36. Thus, we corroborated a chordoma locus on chromosome 7q in the 3 original families and demonstrated evidence for genetic heterogeneity in the fourth family. Our study also provided insights into some limitations and analytical complexities associated with using a dense SNP marker set in linkage analysis of complex pedigrees. PMID- 15818628 TI - High-resolution NMR spectroscopy in solids by truly magic-angle spinning. PMID- 15818629 TI - Birnbaumin A and B: two unusual 1-hydroxyindole pigments from the "flower pot parasol" Leucocoprinus birnbaumii. PMID- 15818630 TI - Molecular recognition by calix[4]arene-modified gold nanoparticles in aqueous solution. PMID- 15818631 TI - Rational design of a coordination cage with a trigonal-bipyramidal shape constructed from 33 building units. PMID- 15818632 TI - Nanoporous polyelectrolyte spheres prepared by sequentially coating sacrificial mesoporous silica spheres. PMID- 15818634 TI - The role of competency to stand trial in mental health courts. AB - Concerns have been raised in the literature about the competency to stand trial and competency to make treatment decisions of defendants referred to mental health courts. However, there is little information reported about the evaluation and prevalence of incompetence, characteristics of incompetent mental health court defendants, and disposition of mentally ill defendants too disturbed to be diverted from the criminal justice system through mental health courts. This study reports on the 85 potential mental health court defendants referred for trial competency evaluations during the first three years of operation of the Akron Ohio Mental Health Court. Of the 80 defendants who could be located for evaluation, 77.5% were found incompetent, and 53% of the incompetent defendants were not restored to competence even after an average of 49 days of treatment in a state psychiatric hospital. The implications of these findings in terms of the diversion potential of mental health courts for the severely mentally ill are discussed. PMID- 15818635 TI - What is the role of muscle receptors in proprioception? AB - The role of muscle afferents is discussed in terms of their contribution to kinesthesia, the senses of position and movement of the limbs. It is argued that muscle spindles are not well suited as position sensors, on several grounds. Yet we know from muscle vibration experiments that they do contribute to kinesthesia. A number of recent experiments have shown that positional information is of particular significance to the central nervous system. In other experiments it has been demonstrated that a disturbance to kinesthesia follows fatigue from exercise. Fatigue of elbow flexor muscles led subjects to make significant positional errors in a forearm matching task. The size of the errors correlated with the fall in force from fatigue. These data suggest that we derive a positional cue from the effort required to hold a limb against the force of gravity. A challenge for the future will be to reveal how the centrally derived sense of effort and peripherally derived afferent information interact to give us our kinesthetic sense. PMID- 15818636 TI - Communicating safety information to physicians: an examination of dear doctor letters. AB - PURPOSE: Recent research suggests that letters to healthcare providers may not effectively communicate safety-related information. This study examined whether content, organization, and formatting of dear doctor letters (DDLs) influences physicians' responses to the letters. METHODS: Drugs with warning label changes during 2000 and 2001 were identified, and corresponding DDLs were obtained. Letters were coded for content, organization, and formatting. Ten physicians rated letters on presentation, criticalness of the information, and likelihood of changing practice as a result of the letter. Areas of deficiency were identified. The relationships between key characteristics of the letters and physicians' ratings were examined using correlations; reliability of physicians' ratings was estimated using generalizability theory. RESULTS: For 2000 and 2001, 124 drugs were identified as having had changes to the warning section of the label; DDLs were sent in 32 (25.8%) instances. Letters varied in terms of the placement of key information, use of formatting, and length. Physicians' ratings suggested 25% of the letters were deficient in clarity, 28% in readability, 36% in the ratio of relevant information to supporting information, 36% in key information easily discernable, and 28% in overall effectiveness of communication. Letters with formatting highlighting key information were preferred. Letter length and placement of key information were not correlated with physicians' ratings. CONCLUSIONS: Many DDLs do not communicate labeling changes clearly and effectively. If DDLs are used to communicate safety information to physicians, special formatting and explicit wording should be used to emphasize new information. PMID- 15818637 TI - Regional variation in the prescribing for diabetes and use of secondary preventative therapies in Ireland. AB - PURPOSE: To compare the prescribing of secondary preventative therapies for patients with both insulin-dependent diabetes mellitus (IDDM) and non-insulin dependent diabetes mellitus (NIDDM) in the eight health board regions of Ireland. METHODS: We utilized data from the national general medical services (GMS) prescribing database to examine the variability of prescribing for diabetes and associated secondary therapies between regions in those aged 45 years or more. Age-sex standardized prescribing rates of six secondary preventative therapies (aspirin, beta-blockers, statins, ACE inhibitors, angiotensin receptor (AT2) antagonists, and fibrates) were calculated for each region. RESULTS: Variations exist between regions for treated NIDDM (1.5-fold) and IDDM (1.5-fold). Wide variations were observed between regions for prescribing of secondary preventative therapies with the highest variability observed for statin prescribing (1.5- to 1.6-fold) and for AT2 antagonist prescribing (2.0-fold) in NIDDM patients. In those with NIDDM, men were more likely to receive aspirin OR=1.26 (1.21--1.31), ACE inhibitors 1.14 (1.101.18), and fibrates OR=1.55 (1.23- 1.96) than women and those aged over 75 years were less likely to receive statins OR=0.60 (0.56--0.65) and fibrates OR=0.25 (0.17--0.37) than those aged 45--74. Similar results were also shown for patients with IDDM. CONCLUSIONS: The results suggest that access to secondary preventative therapy in diabetes patients is not equitable across regions, gender, and age in Ireland. While much of the variability remains unexplained, it may be due to differences in screening and health promotion between regions, prescriber uncertainty, variability in clinical need, or may be derived from a socioeconomic disparity among regions. PMID- 15818638 TI - Antimicrobial prescribing at a university hospital: justified or 'just in case': testing a new scoring system as a key quality indicator. AB - OBJECTIVE: To evaluate the quality of antimicrobial drug prescribing at a university hospital in the Department of Medicine, by using a new scoring system as a quality indicator. METHODS: DESIGN: a prospective, longitudinal survey, during a 21-week period. The necessity of antimicrobial treatment of all in patients at the Department of Medicine, to whom new antimicrobials were prescribed, was assessed by using a scoring system based on the presence of signs and symptoms of an infection. If the sum's total score was 3 or more, the antimicrobial treatment was deemed justifiable; if the score was less than 3, the antimicrobial treatment was regarded as questionable. SETTING: Department of Medicine (279 hospital-beds) at the University Hospital Center Rijeka, a 1200 patient-bed teaching hospital in Croatia. RESULTS: Antimicrobials were prescribed to 15% of the total patients. They were given as a treatment to 89% of the patients, and in 67% of the cases, this treatment was administered empirically. According to the scoring system, 29% of the patients did not have a justified indication for antimicrobial treatment. CONCLUSION: The proposed quality indicator (scoring system) that we used is a simple method for the quality assessment of antimicrobial use. It has indicated areas that require in-depth analysis. PMID- 15818639 TI - A study of adherence to drug recommendations by providing feedback of outpatient prescribing patterns to hospital specialists. AB - PURPOSE: To study the effect of feedback using prescribing profiles combined with interactive group discussions on hospital specialists' adherence to evidence based guidelines for drug treatment of common diseases issued by a regional Drug and Therapeutics Committee. METHODS: Intervention study performed at 17 clinics at a university hospital in a Swedish metropolitan health region with comparative clinics at a second university hospital as a control. Prescribing profiles based on aggregate pharmacy dispensing data were presented for the physicians in interactive group discussions. Deviations from the guidelines were discussed at each clinic and specific goals of improvement were formulated. The effect was assessed by pre- and post-intervention comparison of the adherence to guidelines for all drugs and within 11 selected therapeutic areas. The credibility and usefulness of the prescribing profiles were evaluated by a questionnaire. RESULTS: The adherence to pharmaceutical products within the pharmacological groups stated in the guideline increased by 2.8%-units at the intervention hospital compared with 0.8%-units at the control hospital. The adherence to drug substance increased by 0.4%-units at the intervention hospital while it decreased by 1.8%-units at the control hospital. For 8 of 11 pre-defined specific goals of improvement, the change in adherence was more positive at the intervention hospital. Most doctors considered the feedback provided clear and relevant. CONCLUSIONS: Interactive group discussions with prescribing profiles were found to be useful in improving hospital specialists' adherence to guidelines. However, the effect on the overall adherence was modest, indicating the importance of clear messages for improvement and relevant guidelines for the prescribing of specialist drugs but also more precise methods for evaluating the effect of real life-interventions. PMID- 15818640 TI - Effectiveness of manual therapy or pulsed shortwave diathermy in addition to advice and exercise for neck disorders: a pragmatic randomized controlled trial in physical therapy clinics. AB - OBJECTIVE: To determine whether manual therapy or pulsed shortwave diathermy, in addition to advice and exercise, provide better clinical outcome at 6 months than advice and exercise alone in primary care patients with nonspecific neck disorders. METHODS: This was a multicenter, 3-arm randomized controlled trial in 15 physical therapy departments. Of the 735 screened patients, 350 were recruited to the study (mean age 51 years) from July 2000 to June 2002. Participants were randomized to advice and exercise plus manual therapy, advice and exercise plus pulsed shortwave, or advice and exercise alone. Assessments were undertaken at baseline, 6 weeks, and 6 months. The primary outcome was the Northwick Park Neck Pain Questionnaire. Analysis was by intention to treat. RESULTS: Of the participants, 115 were allocated to advice and exercise, 114 to advice and exercise plus manual therapy, and 121 to advice and exercise plus pulsed shortwave; 98% received the allocated treatment. There was 93% followup at 6 months. The mean +/- SD fall in Northwick Park score at 6 months was 11.5 +/- 15.7 for advice and exercise alone, 10.2 +/- 14.1 for advice and exercise plus manual therapy, and 10.3 +/- 15.0 for advice and exercise plus pulsed shortwave. There were no statistically significant differences in mean changes between groups. CONCLUSION: The addition of pulsed shortwave or manual therapy to advice and exercise did not provide any additional benefits in the physical therapy treatment of neck disorders. PMID- 15818641 TI - Critical appraisal of qualitative research in clinical journals challenged. PMID- 15818642 TI - Remitting seronegative symmetrical synovitis with pitting edema of the hands: ultrasound, color doppler ultrasound, and magnetic resonance imaging findings. AB - OBJECTIVE: To assess the value of gray-scale ultrasound (US), color Doppler ultrasound (CDUS), contrast-enhanced CDUS, and magnetic resonance imaging (MRI) in the diagnostic evaluation of the hands in patients with remitting seronegative symmetrical synovitis with pitting edema (RS3PE). METHODS: Eight patients (5 men, 3 women; mean +/- SD age 69.3 +/- 7.2 years) with clinical diagnosis of RS3PE syndrome underwent US, CDUS, contrast-enhanced CDUS, and MRI. US was performed with a linear array transducer operating at 12 MHz. The US contrast agent (SHU 508; Levovist, Schering, Germany) was intravenously infused in a concentration of 300 mg/ml at a rate of 1 ml/minute. RESULTS: All patients showed symmetric subcutaneous edema and synovitis of tendons and finger joints on both US and MRI. Vascularity was detected subcutaneously in tendon sheaths and in the joint synovia on CDUS and MRI. Detection of increased vascularity was improved after contrast administration compared with unenhanced CDUS (P < 0.01). CONCLUSION: Ultrasound, CDUS, contrast-enhanced CDUS, and MRI are valuable tools in the diagnostic evaluation of involved anatomic structures in patients with RS3PE. Contrast-enhanced CDUS is superior to CDUS in assessment of inflammatory edema, effusion, and synovitis. PMID- 15818643 TI - Magnetic resonance imaging in optic neuritis due to giant cell arteritis. PMID- 15818644 TI - Redesigning the care of fragility fracture patients to improve osteoporosis management: a health care improvement project. AB - OBJECTIVE: To develop new processes that assure more reliable, population-based care of fragility fracture patients. METHODS: A 4-year clinical improvement project was performed in a multispecialty, community practice health system using evidence-based guidelines and rapid cycle process improvement methods (plan-do study-act cycles). RESULTS: Prior to this project, appropriate osteoporosis care was provided to only 5% of our 1999 hip fracture patients. In 2001, primary physicians were provided prompts about appropriate care (cycle 1), which resulted in improved care for only 20% of patients. A process improvement pilot in 2002 (cycle 2) and full program implementation in 2003 (cycle 3) have assured osteoporosis care for all willing and able patients with any fragility fracture. Altogether, 58% of 2003 fragility fracture patients, including 46% of those with hip fracture, have had a bone measurement, have been assigned to osteoporosis care with their primary physician or a consultant, and are being monitored regularly. Only 19% refused osteoporosis care. Key process improvements have included using orthopedic billings to identify patients, referring patients directly from orthopedics to an osteoporosis care program, organizing care with a nurse manager and process management computer software, assigning patients to primary or consultative physician care based on disease severity, and monitoring adherence to therapy by telephone. CONCLUSION: Reliable osteoporosis care is achievable by redesigning clinical processes. Performance data motivate physicians to reconsider traditional approaches. Improving the care of osteoporosis and other chronic diseases requires coordinated care across specialty boundaries and health system support. PMID- 15818645 TI - Comprehensive assessment of clinical outcome and quality of life after resection interposition arthroplasty of the thumb saddle joint. AB - OBJECTIVE: To explore the biometric and psychometric properties of clinical, generic, and condition-specific instruments and to assess quantitatively the outcome after resection interposition arthroplasty (RIAP) of the thumb saddle joint. METHODS: One hundred three patients requiring 112 arthroplasties were assessed in a 4.5-7.7-year cross-sectional catamnesis by means of 4 widely used questionnaires and clinical and radiographic examinations. RESULTS: In all dimensions of the Short Form 36 (SF-36), the outcome was equal or significantly better than expected by the norm. The Disability of the Arm, Shoulder and Hand questionnaire (DASH) revealed some small, mainly functional limitations (mean score 78.4, norm 86.4). The SF-36, the DASH, and the Patient Related Wrist Evaluation form (PRWE) correlated highly and loaded on the same factor. The Hand Function Index was independent of the clinical measurements (range of motion, strength, etc. on the specially designed Custom Form) and of the self rating. CONCLUSION: Long-term followup of 112 RIAP patients showed excellent health and quality of life. A questionnaire set consisting of the SF-36, the DASH (or alternatively the short PRWE), and the Custom Form is proposed for the comprehensive and specific assessment of thumb joint conditions. PMID- 15818646 TI - Relationship of pain-coping strategies and pain-specific beliefs to pain experience in children with juvenile idiopathic arthritis. AB - OBJECTIVE: To examine whether pain-specific beliefs and coping strategies of patients with juvenile idiopathic arthritis (JIA) independently predict their reported pain, while controlling for relevant demographic variables, disease activity, and parent-rated disability. To compare use of pain-coping strategies and pain-related beliefs of a selected subgroup of patients with high pain and low disease activity (high pain group) with the remaining patients. METHODS: Children with JIA (n = 56) completed the Pain Coping Questionnaire, a revised version of the Survey of Pain Attitudes, and a 3-week pain diary. The parents completed the Childhood Health Assessment Questionnaire (CHAQ). Second order principal component analyses were conducted in order to reduce the number of independent variables. Regression analyses of the dependent measure were performed. The use of coping strategies and health beliefs were compared using t tests for independent samples. Pearson's correlation coefficients were calculated to examine the direct associations between each individual coping and belief scale, and the pain measure. RESULTS: Only the CHAQ and the cognitive belief composite factor score made statistically significant contribution to the prediction of pain after controlling for other variables. Significant differences were found between the scores of high pain patients and the rest of the group for the health belief subscale of disability (mean +/- SD 2.0 +/- 0.6 and 1.2 +/- 0.7, respectively), and for the health belief subscale of harm (mean +/- SD 2.7 +/- 0.6 and 1.8 +/- 0.7, respectively). Significant correlations were obtained between the pain measure and the pain-coping subscale of catastrophizing, the pain belief subscales of disability, harm, solicitude (inverse), control, and medical cure. CONCLUSION: These results support a model of pain experience in patients with JIA where psychological factors are strongly influential. It may be efficient to focus behavioral interventions on a subgroup of children where the pain experience seems to be in discordance with the disease activity. PMID- 15818647 TI - Minimizing complications from nonsteroidal antiinflammatory drugs: cost effectiveness of competing strategies in varying risk groups. AB - OBJECTIVE: To appraise the cost-effectiveness of competing therapeutic strategies in patient cohorts eligible for aspirin prophylaxis with varying degrees of gastrointestinal (GI) and cardiovascular risk. METHODS: Cost-effectiveness and cost-utility analyses were performed to evaluate 3 competing strategies for the management of chronic arthritis: 1) a generic nonselective nonsteroidal antiinflammatory drug (NSAID(NS)) alone; 2) NSAID(NS) plus a proton pump inhibitor (PPI); and 3) a cyclooxygenase 2-selective inhibitor (coxib) alone. Cost estimates were from a third-party payer perspective. The outcomes were incremental cost per ulcer complication avoided and incremental cost per quality adjusted life year (QALY) gained. Sensitivity analysis was performed to evaluate the impact of varying patient GI risks and aspirin use. RESULTS: In average-risk patients, the NSAID(NS) + PPI strategy costs an incremental 45,350 US dollars per additional ulcer complication avoided and 309,666 US dollars per QALY gained compared with the NSAID(NS) strategy. The coxib strategy was less effective and more expensive than the NSAID(NS) + PPI strategy. Sensitivity analysis revealed that the NSAID(NS) + PPI strategy became the dominant approach in patients at high risk for an NSAID adverse event (i.e., patients taking aspirin with > or =1 risk factor for a GI complication). CONCLUSION: Generic nonselective NSAIDs are most cost-effective in patients at low risk for an adverse event. However, the addition of a PPI to a nonselective NSAID may be the preferred strategy in patients taking aspirin or otherwise at high risk for a GI or cardiovascular adverse event. PMID- 15818648 TI - Opportunistic infections in polymyositis and dermatomyositis. AB - OBJECTIVE: To assess prevalence and characteristics of opportunistic infections in patients with polymyositis/dermatomyositis (PM/DM). To determine the predictive values for opportunistic infections on clinical presentation, biochemical findings, and paraclinical features of PM/DM to detect patients at risk of opportunistic infections. METHODS: The medical records of 156 consecutive PM/DM patients in 3 medical centers were reviewed. RESULTS: Eighteen PM/DM patients (11.5%) developed opportunistic infections. The majority of patients exhibited an opportunistic infection after the onset of PM/DM (89% of cases). Opportunistic infections occurred most frequently during the first year following PM/DM diagnosis (62.5%). The pathogen microorganisms responsible for opportunistic infections were various, i.e., Candida albicans, Pneumocystis carinii, Aspergillus fumigatus, Geotrichum capitatum, Mycobacterium avium intracellulare complex, M. xenopi, M. marinum, M. tuberculosis, Helicobacter heilmanii, cytomegalovirus, and herpes simplex virus. Mortality rates were as high as 27.7% in these PM/DM patients. Higher mean daily doses of steroids, lymphopenia, and lower serum total protein levels were significantly more frequent in the group of PM/DM patients with opportunistic infections. CONCLUSION: Our study underscores the high frequency of opportunistic infections in PM/DM, resulting in an increased mortality rate. It also indicates that a great variety of microorganisms are responsible for opportunistic infections, although they were more often due to fungi (>50% of cases). Our series highlights a predominance of both lung and digestive opportunistic infections (89% of cases). In addition, our results suggest that PM/DM patients presenting with factors predictive of opportunistic infection may require closer monitoring. PMID- 15818649 TI - Positron emission tomography (PET): evaluation of chronic periaortitis. AB - OBJECTIVE: To evaluate the presence and extent of large-vessel inflammation in patients with chronic periaortitis (CP) using (18)F-fluorodeoxyglucose-positron emission tomography (FDG-PET). METHODS: A consecutive case series consisting of 7 patients with CP seen over a 3-year period and a control group of 14 patients with malignancy were evaluated with FDG-PET. For every case we selected 2 age- and sex-matched controls who underwent PET imaging for malignancy. The diagnosis of CP was made by means of computed tomography. PET imaging was performed at diagnosis before therapy was started. Measurement of vascular uptake was graded using a 4-point semiquantitative scale. RESULTS: All patients had evidence of grade 2+ or 3+ vascular uptake in the abdominal aorta and/or iliac artery. No controls showed vascular uptake greater than 1+. Vascular uptake in the thoracic aorta and/or in its branches was seen in 3 (43%) of 7 patients. Vascular uptake in abdominal aorta and/or iliac artery was observed in patients with CP but not in controls (100% versus 0%). There was also a significantly more frequent FDG uptake in the large thoracic arteries in case-patients compared with controls (43% versus 0%; P = 0.03). CONCLUSION: FDG-PET scan shows in patients with CP the presence of a large-vessel vasculitis involving abdominal aorta and common iliac arteries, which in some patients is also extended to thoracic aorta and/or its branches. PMID- 15818650 TI - Two major spondylarthropathy phenotypes are distinguished by pattern analysis in multiplex families. AB - OBJECTIVE: To examine whether spondylarthropathy (SpA) disease manifestations would combine in any ordered pattern among patients from SpA multiplex families. METHODS: SpA patients (n = 540) belonging to 190 multiplex families were thoroughly investigated. Clinical data was collected, systematic pelvic radiographs were taken, and HLA-B27 status was determined. The patterns of SpA manifestations were examined by several methods, including multiple correspondence analysis, nonhierarchical and hierarchical clustering, and discriminant analysis. RESULTS: The nonhierarchical cluster analysis allowed us to classify patients, independent of disease duration, into 2 major groups of comparable size. Group A contained a majority of the women, whereas group B predominantly consisted of men. The 2 groups were very similar regarding axial symptoms, radiographic sacroiliitis, and uveitis. Group B was characterized by a younger age at onset and a higher frequency of clinical enthesitis, peripheral arthritis, dactylitis, psoriasis, and inflammatory bowel disease than group A. Patients belonging to those groups exhibited some degree of familial aggregation, thereby supporting their intrinsic validity. CONCLUSION: Pattern analysis of SpA manifestations among familial SpA allowed us to recognize 2 main clusters independent of disease duration. Furthermore, there was a trend toward aggregation by cluster among families, suggesting that they are determined by specific genetic factors. These clusters may indeed correspond to different severity patterns. PMID- 15818651 TI - Clinical academic rheumatology: getting more than you pay for. AB - OBJECTIVE: Rheumatology is among the least compensated specialties in medicine today. This is a significant problem for clinical rheumatologists in academic medicine who are often expected to earn their salaries through clinical practice alone. Additionally, academic rheumatologists usually cannot generate revenue through office laboratory monitoring, radiographs, or bone densitometry to supplement their income (i.e., downstream income). The purpose of our study was to examine revenue generated from downstream income to a university by a clinical academic rheumatologist. METHODS: Consecutive outpatients (n = 127) seen predominantly by one academic rheumatologist over one month of clinic were followed for 18 months. The total physician compensation for patient visits was calculated and compared with the revenue generated from laboratory tests, radiologic studies, consultations, and specific rheumatologic treatments and procedures performed or ordered. Medicare reimbursement rates for 2003 were used as compensation standards for all charges. RESULTS: Physician office visit billing generated 36,297 US dollars from 730 office visits. The total amount of downstream income from these office visits was 363,813 US dollars (47,386 US dollars from laboratory tests, 35,582 US dollars from radiologic studies, 8,159 US dollars from rheumatologic procedures, 261,584 from rheumatologic infusions, and 11,101 US dollars from initial consultations). Therefore, 10.02 US dollars of downstream revenue was generated for every 1.00 of office visit compensation applied to the academic rheumatologist's salary. CONCLUSION: Although academic rheumatologists struggle to bill their salaries through seeing more patients, they are clearly a bargain for a university hospital because they generate >10.00 US dollars for every 1.00 US dollars they receive for an office visit. PMID- 15818652 TI - Impact of ultrasound imaging on local corticosteroid injections of symptomatic ankle, hind-, and mid-foot in chronic inflammatory diseases. AB - OBJECTIVE: To assess the impact of ultrasonography (US) on local corticosteroid (CS) injections of painful ankle, hindfoot, and midfoot in chronic inflammatory diseases. METHODS: Consecutive patients with chronic rheumatic diseases admitted to the hospital for local CS injections of painful ankle, hindfoot, or midfoot were enrolled in this study. Clinical and radiographic evaluation was performed by the physician in charge of CS injections, and US examination was performed by an independent examiner blinded to the clinical and radiologic findings. According to a randomized weekly-periods design, the physician planned CS injections either aware (G1 group) or unaware (G2 group) of US results. In the latter case, he was nonetheless informed of US results after he had performed the injections. Impact of this information on the treatment planning was assessed in all cases. Prognostic impact of US was also evaluated by comparing the change in global assessment of efficacy of CS injections, in activity of the disease, and in the Western Ontario and McMaster Universities (WOMAC) subscales after 1 and 3 months, between G1 and G2 groups. RESULTS: The knowledge of US findings led the physician to change his decision of local CS injections in 56 (82%) of 68 patients studied. Among 1,131 assessed sites, by clinical, radiographic, and US evaluation, injection was cancelled in 37 (15%) of 242 proposed sites, whereas it was decided in 74 (8%) additional sites. After 1 month, there was an improvement in G1 as compared with G2 groups. The mean +/- SD change in WOMAC physical function subscale was 15.6 +/- 17.5 in G1 versus 8 +/- 13 in G2 (P = 0.0305). After 3 months, only the global assessment of efficacy of CS injections was statistically greater in G1 than in G2 group (P = 0.0170). CONCLUSION: US frequently led the physician to change his diagnosis of inflammatory lesions in painful foot, and consequently his planning of CS injections. Moreover, US could improve the response to local treatment. PMID- 15818653 TI - Determinants of sleep quality in women with systemic lupus erythematosus. AB - OBJECTIVE: To characterize sleep complaints in women with systemic lupus erythematosus (SLE) and to identify correlates of sleep quality. METHODS: Sleep quality in 100 women with SLE was assessed using the Pittsburgh Sleep Quality Index (PSQI). Participants completed standardized questionnaires assessing depressed mood, leisure time physical activity, functional disability, and pain severity. A clinical examination determined disease activity, cumulative damage, and whether patients fulfilled the American College of Rheumatology criteria for fibromyalgia. A series of hierarchical multiple regressions were computed. RESULTS: The mean +/- SD global PSQI score was 6.98 +/- 4.03, with moderate to severe sleep impairment reported by 56% of the sample. The first model testing the importance of demographic factors was not statistically significant. In the disease-related model, the use of prednisone and functional disability both contributed to poor sleep quality (P < 0.001). The addition of level of exercise participation to the demographic set significantly added to the model (P = 0.001). Depression significantly added to the demographic set, explaining 29% of the variance (P < 0.0001). When these variables, along with disease related variables, were simultaneously regressed on the PSQI Global Score, only depressed mood appeared as a significant independent determinant of global sleep quality (P < 0.001). However, the point estimates for the Beta coefficients were consistent with effects for lack of exercise and prednisone use. CONCLUSION: A significant proportion of women with SLE suffer from poor sleep quality. The findings suggest that depressed mood, prednisone use, and lack of exercise contribute to decreased overall sleep quality. PMID- 15818654 TI - History of infection before the onset of juvenile dermatomyositis: results from the National Institute of Arthritis and Musculoskeletal and Skin Diseases Research Registry. AB - OBJECTIVE: To obtain data concerning a history of infection occurring in the 3 months before recognition of the typical weakness and rash associated with juvenile dermatomyositis (JDM). METHODS: Parents or caretakers of children within 6 months of JDM diagnosis were interviewed by the registry study nurse concerning their child's symptoms, environment, family background, and illness history. Physician medical records were reviewed, confirming the JDM diagnosis. RESULTS: Children for which both a parent interview and physician medical records at diagnosis were available (n = 286) were included. Diagnoses were as follows: definite/probable JDM (n = 234, 82%), possible JDM (n = 43, 15%), or rash only (n = 9, 3%). The group was predominantly white (71%) and had a girl:boy ratio of 2:1. Although the mean age at onset was 6.7 years for girls and 7.3 years for boys, 25% of the children were < or =4 years old at disease onset. In the 3 months before onset, 57% of the children had respiratory complaints, 30% had gastrointestinal symptoms, and 63% of children with these symptoms of infection were given antibiotics. CONCLUSION: This study provides evidence that JDM affects young children. The symptoms of the typical rash and weakness often follow a history of respiratory or gastrointestinal complaints. These data suggest that the response to an infectious process may be implicated in JDM disease pathogenesis. PMID- 15818655 TI - Gaps in care for rheumatoid arthritis: a population study. AB - OBJECTIVE: Treatment guidelines for rheumatoid arthritis (RA) now recommend early, aggressive, and persistent use of disease-modifying antirheumatic drugs (DMARDs) to prevent joint damage in all people with active inflammation, and evaluation by a rheumatologist, when possible. This research assesses whether care for RA, at a population level, is consistent with current treatment guidelines. METHODS: Using administrative billing data from the Ministry of Health in 1996-2000, all prevalent RA cases in British Columbia, Canada were identified. Data were obtained on all medications and all provincially-funded health care services. RESULTS: We identified 27,710 RA cases, yielding a prevalence rate of 0.76%, consistent with epidemiologic studies. DMARD use was inappropriately low. Only 43% of the entire RA cohort received a DMARD at least once over 5 years, and 35% over 2 years. When used, DMARDs were started in a timely fashion, but were not used consistently. Care by a rheumatologist increased DMARD use 31-fold. Yet, only 48% and 34% saw a rheumatologist over 5 and 2 years, respectively. DMARD use was significantly more frequent, persistent, and more often used as combination therapy with continuous rheumatologist care. DMARDs were used by 84% and 73%, 40%, and 10% of people followed by rheumatologists continuously and intermittently, internists, and family physicians, respectively (P < 0.001). NSAID use, physiotherapy, and orthopedic surgeries were similar across these 4 care groups. CONCLUSION: RA care in the British Columbia population was not consistent with current treatment guidelines. Efforts to educate family physicians and consumers about the shift in RA treatment paradigms and to improve access to rheumatologists are needed. PMID- 15818656 TI - Are health care professionals advising adults with arthritis to become more physically active? AB - OBJECTIVE: To describe the proportion and characteristics of adults with self reported, doctor-diagnosed arthritis who report ever having been advised by a health professional to become more physically active and to assess whether the advice was associated with recent physical activity. METHODS: Using population based data from the 2003 Behavioral Risk Factor Surveillance Survey, respondents were classified according to their arthritis status, whether or not they were ever advised by a health professional to increase their physical activity to help them better manage their arthritis, and whether they engaged in exercise or physical activity within the past month. RESULTS: Overall, 42% of respondents with arthritis reported ever being advised to increase their physical activity to help their arthritis. Respondents who were more likely to have ever received the advice were female, middle-aged or older, African American, Hispanic, overweight or obese, sedentary or insufficiently physically active, and had higher levels of education. Persons who reported ever receiving the advice were less likely to report that they had exercised within the past month. A higher level of education was the only variable associated with recent exercise or physical activity among those advised to be more active. CONCLUSION: Less than 50% of adults with arthritis report ever being advised by a health professional to become more physically active. Advice alone appears insufficient to promote increased physical activity in adults with arthritis. PMID- 15818657 TI - Effects of supervised aerobic exercise in patients with systemic lupus erythematosus: a pilot study. PMID- 15818658 TI - Validity of the 6-minute walking test in juvenile idiopathic arthritis. PMID- 15818659 TI - Fibroblast expression of the coactivator p300 governs the intensity of profibrotic response to transforming growth factor beta. AB - OBJECTIVE: Transforming growth factor beta (TGFbeta) induces profibrotic responses in normal fibroblasts, and plays a fundamental role in the pathogenesis of fibrosis in scleroderma (systemic sclerosis [SSc]). The intensity of cellular responses elicited by cytokines is modulated by transcriptional coactivators such as the histone acetylase p300. The objective of these studies was to delineate the physiologic role of p300 in Smad-dependent profibrotic responses elicited by TGFbeta. METHODS: Ectopic p300 was transiently expressed in normal dermal fibroblasts. Cellular p300 levels were suppressed using p300-specific ribozymes. The regulation of gene expression was examined by transient transfection assays, Northern blotting, and immunoblot analysis. The expression of p300 in normal and scleroderma fibroblasts was evaluated by confocal microscopy and immunoblotting, and p300 levels in skin from mice with experimental scleroderma were assessed by immunohistochemistry. RESULTS: In normal fibroblasts, TGFbeta induced an increase in the levels of p300. Forced expression of ectopic p300 in these cells dramatically enhanced the magnitude of TGFbeta responses, whereas selective depletion of p300 using ribozyme resulted in abrogation of TGFbeta-induced collagen synthesis and promoter activity. Furthermore, TGFbeta lost its ability to induce Smad-dependent transcription in p300-depleted fibroblasts. These responses could be fully rescued with ectopic p300. Abrogation of Smad-mediated TGFbeta signaling was not due to alterations in the levels or the ligand dependent phosphorylation or intracellular trafficking of endogenous Smads. Immunohistochemical analysis demonstrated substantially increased p300 expression in lesional skin from mice with chronic graft-versus-host disease, an animal model of scleroderma. Furthermore, levels of p300 were 2-3-fold higher in cultured fibroblasts derived from SSc patients than in fibroblasts from matched normal controls. CONCLUSION: These results establish, for the first time, that the coactivator histone acetylase p300, itself a target of TGFbeta regulation, is an essential component of the cellular TGFbeta signal transduction pathways mediating stimulation of collagen synthesis in fibroblasts. Since the cellular abundance of p300 appears to govern the intensity of profibrotic responses elicited by TGFbeta, elevated p300 expression in lesional tissue may contribute to the progression of skin fibrosis in scleroderma. PMID- 15818660 TI - Moraxella catarrhalis-induced septic arthritis of a prosthetic knee joint in a patient with rheumatoid arthritis treated with anakinra: comment on the article by Schiff et al. PMID- 15818661 TI - Pain, stiffness, and fatigue in juvenile polyarticular arthritis: contemporaneous stressful events and mood as predictors. AB - OBJECTIVE: To analyze patterns of stress, mood, disease symptoms, and activity reduction in children with polyarticular arthritis, using a prospective daily diary method. METHODS: For a 2-month period, 51 children with polyarticular arthritis (mean age 12.4 years, 65% female) completed daily diaries that included measures of symptoms of pain, stiffness, and fatigue, as well as stress, mood, and activity reduction. Functional status and disease activity were assessed at the initial and followup evaluations with use of the Childhood Health Assessment Questionnaire, physician global assessment, joint count, and laboratory testing. RESULTS: Children reported having pain, stiffness, and fatigue on >70% of days, with significant variability in symptom levels. Results revealed significant same day relationships between stress, mood, and disease symptoms, after controlling for covariates. Specifically, daily fluctuations in both stress and mood were predictive of increased pain, stiffness, and fatigue. Increases in daily stress, mood, and disease symptoms were also significantly related to decreased participation in social activities on a day-to-day basis. Only mood and stiffness were predictors of a cutback in school attendance. CONCLUSION: Stress and mood are important predictors of daily disease symptoms in children with polyarticular arthritis. Moreover, daily fluctuations in stress, mood, and disease symptoms are predictive of aspects of daily function, including participation in school and social activities. Thus, health care providers should solicit daily symptom reports when making decisions regarding clinical management. Nonpharmacologic interventions such as cognitive-behavioral therapy and stress management may be useful adjuvant therapy when treating the disease symptoms of children with polyarticular arthritis. PMID- 15818662 TI - Differential effects of the immunosuppressant FK-506 on human alpha2(I) collagen gene expression and transforming growth factor beta signaling in normal and scleroderma fibroblasts. AB - OBJECTIVE: To investigate the effects of FK-506 on the expression of the human alpha2(I) collagen gene and transforming growth factor beta (TGFbeta) signaling in normal and scleroderma fibroblasts. METHODS: The expression levels of type I procollagen protein and alpha2(I) collagen messenger RNA (mRNA) were analyzed by immunoblotting and Northern blotting, respectively. The promoter activities of alpha2(I) collagen gene and 3TP-Lux were determined by transient transfection assay. Interaction between TGFbeta receptor type I and FK-506 binding protein 12 (FKBP12) was evaluated by immunoprecipitation. RESULTS: FK-506 did not affect the basal expression of type I procollagen protein or alpha2(I) collagen mRNA, but it significantly reduced the TGFbeta1-induced expression of type I procollagen protein and alpha2(I) collagen mRNA in normal fibroblasts. The effect of FK-506 was regulated posttranscriptionally, but not transcriptionally. In scleroderma fibroblasts, FK-506 significantly reduced the expression of type I procollagen protein and alpha2(I) collagen mRNA through posttranscriptional regulation, but not transcriptional regulation. FK-506 increased the basal activity of the 3TP Lux promoter, but it did not affect the TGFbeta1-induced promoter activity in normal fibroblasts. In contrast, FK-506 did not affect the basal or the TGFbeta1 induced 3TP-Lux promoter activity in scleroderma fibroblasts. Furthermore, FKBP12, which protects TGFbeta receptor type I from ligand-independent activation by TGFbeta receptor type II, constitutively dissociated from TGFbeta receptor type I in scleroderma fibroblasts. CONCLUSION: FK-506 inhibits alpha2(I) collagen gene expression by reducing the stability of mRNA without exhibiting its activation effect on TGFbeta signaling in scleroderma fibroblasts. PMID- 15818663 TI - New classification of HLA-DRB1 alleles supports the shared epitope hypothesis of rheumatoid arthritis susceptibility. AB - OBJECTIVE: The shared epitope hypothesis was formulated to explain the involvement of HLA-DRB1 in rheumatoid arthritis (RA). However, several studies, which considered only the HLA-DRB1 alleles shown to be associated with RA risk, rejected this hypothesis. In this report, we propose that a different classification of HLA-DRB1 alleles be considered, based on the amino acid sequence at position 70-74. METHODS: The fit of both HLA-DRB1 classifications was tested in 2 groups of RA patients. All subjects were recruited through the European Consortium on Rheumatoid Arthritis Families, and included 100 patients with isolated RA and 132 patients with at least 1 affected sibling. RESULTS: The new classification produced risk estimates that fit all of the observed data, i.e., the distribution of the HLA-DRB1 genotype in the 2 patient groups, and the distribution of parental alleles shared by affected sibpairs. The risk of developing RA under this new classification depends on whether the RAA sequence occupies position 72-74 but is modulated by the amino acid at position 71 (K confers the highest risk, R an intermediate risk, A and E a lower risk) and by the amino acid at position 70 (Q or R confers a higher risk than D). CONCLUSION: A new classification based on amino acid sequence allows us to show that the shared epitope RAA sequence at position 72-74 explains the data, with the risk of developing RA modulated by the amino acids at positions 70 and 71. PMID- 15818664 TI - Association of sporadic chondrocalcinosis with a -4-basepair G-to-A transition in the 5'-untranslated region of ANKH that promotes enhanced expression of ANKH protein and excess generation of extracellular inorganic pyrophosphate. AB - OBJECTIVE: Certain mutations in ANKH, which encodes a multiple-pass transmembrane protein that regulates inorganic pyrophosphate (PPi) transport, are linked to autosomal-dominant familial chondrocalcinosis. This study investigated the potential for ANKH sequence variants to promote sporadic chondrocalcinosis. METHODS: ANKH variants identified by genomic sequencing were screened for association with chondrocalcinosis in 128 patients with severe sporadic chondrocalcinosis or pseudogout and in ethnically matched healthy controls. The effects of specific variants on expression of common markers were evaluated by in vitro transcription/translation. The function of these variants was studied in transfected human immortalized CH-8 articular chondrocytes. RESULTS: Sporadic chondrocalcinosis was associated with a G-to-A transition in the ANKH 5' untranslated region (5'-UTR) at 4 bp upstream of the start codon (in homozygotes of the minor allele, genotype relative risk 6.0, P = 0.0006; overall genotype association P = 0.02). This -4-bp transition, as well as 2 mutations previously linked with familial and sporadic chondrocalcinosis (+14 bp C-to-T and C-terminal GAG deletion, respectively), but not the French familial chondrocalcinosis kindred 143-bp T-to-C mutation, increased reticulocyte ANKH transcription/ANKH translation in vitro. Transfection of complementary DNA for both the wild-type ANKH and the -4-bp ANKH protein variant promoted increased extracellular PPi in CH-8 cells, but unexpectedly, these ANKH mutants had divergent effects on the expression of extracellular PPi and the chondrocyte hypertrophy marker, type X collagen. CONCLUSION: A subset of sporadic chondrocalcinosis appears to be heritable via a -4-bp G-to-A ANKH 5'-UTR transition that up-regulates expression of ANKH and extracellular PPi in chondrocyte cells. Distinct ANKH mutations associated with heritable chondrocalcinosis may promote disease by divergent effects on extracellular PPi and chondrocyte hypertrophy, which is likely to mediate differences in the clinical phenotypes and severity of the disease. PMID- 15818666 TI - Uncoupling of inflammation and destruction in rheumatoid arthritis: myth or reality? PMID- 15818667 TI - Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis. AB - OBJECTIVE: RNA interference is a process in which genes can be silenced sequence specifically. In mammals, RNA interference can be invoked by introduction of small (19-21-nucleotide) double-stranded RNA molecules known as small interfering RNA (siRNA) into cells. Thereby, siRNA offers promise as a novel therapeutic modality. However, siRNA is a relatively large, highly charged molecule and does not readily enter cells. This study was undertaken to investigate the use of electroporation for in vivo transfection of siRNA into joint tissue in arthritic mice to achieve local RNA interference. METHODS: Proof of principle that siRNA is able to inhibit gene expression in vivo in the mouse joint was studied by local injection and electroporation of siRNA designed to silence reporter genes. In mice with collagen-induced arthritis (CIA), the disease-modulating activity of siRNA designed to silence tumor necrosis factor alpha (TNFalpha) was investigated. RESULTS: Luciferase activity could be reduced by >90% with luciferase-specific siRNA as compared with the activity measured after electroporation without siRNA or with irrelevant siRNA. The effect was observed only locally. In mice with CIA, electroporation of siRNA designed to inhibit TNFalpha strongly inhibited joint inflammation, whereas electroporation of irrelevant siRNA or injection of siRNA against TNFalpha without electroporation failed to produce therapeutic effects. CONCLUSION: Local electroporation of siRNA in joint tissue can inhibit CIA in mice. These results offer promise for the use of siRNA as a new strategy for therapeutic intervention in rheumatoid arthritis and may serve as a tool to study arthritis disease pathways through loss-of function phenotypes. PMID- 15818669 TI - Association of the Frizzled-related protein gene with symptomatic osteoarthritis at multiple sites. AB - OBJECTIVE: To confirm the association of 2 variants of the Frizzled-related protein gene (FRZB) with osteoarthritis (OA) of the hip, and to investigate whether these variants also associate with other heritable generalized OA phenotypes. METHODS: An association analysis of 2 variants (R200W and R324G) of FRZB was performed in a random sample of 1,369 subjects (ages 55-70 years) from a population-based cohort (the Rotterdam Study) scored for radiographic characteristics of OA in the hip, hand, spine, and knee and in a patient population of Caucasian probands (ages 40-70 years) and their siblings selected for the presence of primary symptomatic OA at multiple sites. RESULTS: The allele frequency of the 2 variants was not significantly different between subjects with hip radiographic OA (ROA) and controls. The frequency of the G allele of the R324G variant was significantly increased in subjects with generalized ROA from the Rotterdam Study (0.10) and in subjects from the Genetics, osteoARthritis and Progression study (0.11) compared with that in controls from the Rotterdam Study (0.08). Carriers of this G allele had increased susceptibility for generalized ROA (odds ratio [OR] 1.4, 95% confidence interval [95% CI] 0.9-1.9, P = 0.10) or familial symptomatic OA at multiple sites (OR 1.6, 95% CI 1.1-2.3, P = 0.02). CONCLUSION: Our results confirm that the R324G variant of the FRZB gene is involved in OA and indicate a role of this variant in several generalized OA phenotypes. A more extended OA phenotype may indeed be expected from genetic variation in an essential pathway of skeletal development such as Wnt signaling. PMID- 15818670 TI - Innate immunity and arthritis: neutrophil Rac and toll-like receptor 4 expression define outcomes in infection-triggered arthritis. AB - OBJECTIVE: The role of innate immunity in Chlamydia-induced arthritis has not been defined. The purpose of this study was to examine the role of neutrophils in experimental arthritis in mice with targeted elimination of the small GTPases Rac1 and Rac2, as well as the role of Toll-like receptors (TLRs) in this model. METHODS: Arthritis was induced by intraarticular inoculation of synoviocyte packaged Chlamydia trachomatis. The degree of arthritis was assessed according to joint swelling and pathology scores. The persistence of Chlamydia in joints was assessed by immunoassay. The expression of TLR-2 and TLR-4 in neutrophils was detected by semiquantitative reverse transcription-polymerase chain reaction. RESULTS: In the acute phase, wild-type mice developed more severe arthritis than did Rac-deficient mice, with abundant infiltration of neutrophils into the joint. In the chronic phase, the Rac-deficient mice developed more severe arthritis and demonstrated defective clearance of the pathogen from the joint. In vitro stimulation of neutrophils with Chlamydia up-regulated the expression of TLR-4, but not TLR-2, in wild-type mice. However, neutrophils from Rac-deficient mice did not show this up-regulation of TLR-4. Sustained TLR-4 expression in neutrophils was found to be dependent on the expression of Rac. Mice genetically deficient in TLR-4 demonstrated more severe arthritis than did the controls. Thus, Rac expression plays a profound role in infection-triggered arthritis and demonstrates a bimodal influence on the disease process, exacerbating acute joint inflammation but controlling chronic arthritis. Rac deficiency was associated with diminished TLR-4 expression, impaired host clearance of the pathogen, and more severe chronic arthritis. CONCLUSION: In infection-triggered arthritis, innate immunity plays a critical role. Effective host clearance of an arthritogenic pathogen depends on intact expression of Rac and appropriate expression of TLR-4 by neutrophils. PMID- 15818671 TI - Reduced p53 in peripheral blood mononuclear cells from patients with rheumatoid arthritis is associated with loss of radiation-induced apoptosis. AB - OBJECTIVE: Patients with autoimmune disorders exhibit highly reproducible gene expression profiles in their peripheral blood mononuclear cells. This profile includes, at least in part, a collection of underexpressed genes that encode proteins that inhibit cell cycle progression and stimulate apoptosis. We aimed to determine whether this gene expression profile confers functional liability on lymphocytes from patients with rheumatoid arthritis (RA). METHODS: Viability studies in response to a panel of proapoptotic stimuli revealed that T lymphocytes from patients with RA were resistant to gamma radiation-induced apoptosis, a process known to be dependent on p53. To assess p53 function in RA peripheral blood mononuclear cells, baseline levels of p53 protein and TP53 transcript were measured in patients with RA and controls. The cellular p53 response to gamma radiation was also assessed by immunoblotting. RESULTS: Lymphocytes from patients with RA had lower baseline levels of TP53 messenger RNA (mRNA) and p53 protein than did those from control subjects and were deficient in their ability to increase p53 after exposure to gamma radiation. A subgroup of patients with RA had a second biochemical defect characterized by expression of very low baseline levels of checkpoint kinase 2 mRNA and protein. CONCLUSION: We conclude that defects in the expression of TP53 mRNA and, in a subgroup, defects in expression of CHK2 mRNA, lead to severe defects in apoptosis in patients with RA. We hypothesize that this liability may contribute to autoimmunity. PMID- 15818672 TI - A new haplotype of PDCD1 is associated with rheumatoid arthritis in Hong Kong Chinese. AB - OBJECTIVE: The programmed death 1 (PD-1) molecule is a negative regulator of T cells, and a genetic association between PD-1 and systemic lupus erythematosus and rheumatoid arthritis (RA) in Caucasians has been reported. The aim of this study was to investigate the association of PDCD1 polymorphisms and haplotypes with RA in the Chinese population. METHODS: Three single-nucleotide polymorphisms (SNPs), PD-1.1 G/A, PD-1.3 G/A, and PD-1.5 C/T, were genotyped in 180 patients with RA and 647 healthy controls in a case-control association study. Analyses of the association of genotypes and alleles with disease, haplotype construction, and linkage disequilibrium (LD) were performed. RESULTS: We constructed haplotypes with the alleles of markers PD-1.1 G/A and PD-1.5 C/T and found that the GT haplotype was overrepresented in patients with RA (31%) compared with controls (23%) (P = 0.001, odds ratio [OR] 1.54, 95% confidence interval [95% CI] 1.18-1.99). Among GT double homozygotes the risk of RA was increased even further (OR 2.31, 95% CI 1.31-4.08, P = 0.006). We also observed that the AA genotype of SNP PD-1.1 was associated with a decreased risk for developing RA (OR 0.38, 95% CI 0.15-0.99, P = 0.034). No association for SNP PD-1.5 in RA was found, and SNP PD-1.3 was nonpolymorphic in the Chinese population. CONCLUSION: Our results support the involvement of PDCD1 as a susceptibility gene for RA in the Chinese population. PMID- 15818675 TI - Activation of the type I interferon system in primary Sjogren's syndrome: a possible etiopathogenic mechanism. AB - OBJECTIVE: The etiopathogenesis of primary Sjogren's syndrome (SS) is largely unknown. In other autoimmune diseases, type I interferon (IFN) may play a pivotal role by triggering and sustaining the disease process. We therefore aimed to determine whether patients with primary SS had an activated type I IFN system. METHODS: Salivary gland biopsy specimens and sera from patients with primary SS were investigated for the occurrence of IFNalpha-producing cells and measurable IFNalpha levels, respectively. The ability of primary SS sera together with apoptotic or necrotic cells to induce IFNalpha production in normal peripheral blood mononuclear cells was examined. The IFNalpha inducer was characterized, and IFNalpha-producing cells were identified. Clinical data were correlated with the IFNalpha-inducing capacity of primary SS sera. RESULTS: Numerous IFNalpha producing cells were detected in salivary gland biopsy specimens, despite low serum IFNalpha levels. Autoantibodies to RNA-binding proteins, combined with material released by necrotic or late apoptotic cells, were potent inducers of IFNalpha production in plasmacytoid dendritic cells (PDCs). This appeared to be attributable to RNA-containing immune complexes triggering PDCs by means of RNA and interaction with Fcgamma receptor IIa. The IFNalpha-inducing capacity of sera was associated with positive results of a labial salivary gland biopsy (focus score >/=1) and with dermatologic, hematologic, and pulmonary manifestations. CONCLUSION: Patients with primary SS have an activated type I IFN system. Although virus may initiate the production of IFN, the continued IFNalpha synthesis is caused by RNA-containing immune complexes that activate PDCs to prolong IFNalpha production at the tissue level. This IFNalpha promotes the autoimmune process by a vicious circle-like mechanism, with increased autoantibody production and formation of more endogenous IFNalpha inducers. PMID- 15818676 TI - Reduction of arthritis severity in protease-activated receptor-deficient mice. AB - OBJECTIVE: Protease-activated receptor 1 (PAR-1) is the cell surface receptor for thrombin. It is unclear whether thrombin contributes to inflammation other than by effects on coagulation. We investigated the proinflammatory participation of PAR-1 in antigen-induced arthritis (AIA). METHODS: Arthritis was induced by intraarticular injection of methylated bovine serum albumin (mBSA) in preimmunized PAR-1-deficient (PAR-1(-/-)) and wild-type (WT) mice. The disease was assessed after 7 days by histologic examination of knee joints after decalcification and Safranin O/toluidine blue staining. Serum levels of anti-mBSA IgG, interferon-gamma, and interleukin-4 (IL-4) were determined by enzyme-linked immunosorbent assay. T cell proliferation response was determined by measuring the incorporation of (3)H-thymidine. Cytokine messenger RNA (mRNA) expression was detected in synovial tissues and peritoneal cells by real-time polymerase chain reaction. RESULTS: Arthritis severity was significantly reduced in PAR-1(-/-) mice compared with WT mice (P = 0.017). Analysis of individual aspects of joint histology revealed significant reductions in synovial exudates (P < 0.001), cartilage degradation (P < 0.01), and bone damage (P = 0.05) in PAR-1(-/-) mice. Synovial IL-1, IL-6, and matrix metalloproteinase 13 (MMP-13) mRNA was significantly reduced in PAR-1(-/-) mice. The titers of antigen-specific serum anti-mBSA total IgG, IgG1, and IgG2a were significantly reduced, and serum IL-4 was significantly increased in arthritic PAR-1(-/-) mice. In contrast, no difference was observed in antigen-induced T cell proliferation between PAR-1(-/ ) and WT mice. In vitro, thrombin-induced (but not lipopolysaccharide-induced) IL 1, IL-6, and MMP-13 mRNA expression was significantly impaired in PAR-1(-/-) mice compared with WT controls. CONCLUSION: These data demonstrate the requirement of PAR-1 for the expression of AIA, the development of an antigen-specific Ig response, thrombin-induced macrophage cytokine and MMP expression, and the inhibitory effect of PAR-1 on serum IL-4. We conclude that PAR-1 plays a significant role in this model of arthritis. PMID- 15818677 TI - Folic acid and folinic acid supplements and methotrexate therapy: comment on the article by Morgan et al. PMID- 15818678 TI - The challenge of diagnosis and classification in early ankylosing spondylitis: do we need new criteria? PMID- 15818679 TI - Unexpected onset of psoriasis during infliximab treatment: comment on the article by Beuthien et al. PMID- 15818680 TI - Clinical infection with Strongyloides sterocoralis following etanercept use for rheumatoid arthritis. PMID- 15818681 TI - Bilateral subchondral insufficiency fracture of the femoral head after renal transplantation: a case report. AB - This report describes the case of a 47-year-old woman who was found to have subchondral insufficiency fractures in both femoral heads after renal transplantation. Initially, plain radiographs showed no obvious changes, but magnetic resonance imaging (MRI) revealed an irregular, discontinuous, low intensity band on the T1-weighted image of both hips. The patient was treated nonsurgically. Ten months after onset, the pain in both hips disappeared, and plain radiographs and MRI showed no abnormalities. Subchondral insufficiency fracture should be considered in the diagnosis of patients who have hip pain after renal transplantation. PMID- 15818682 TI - An opposing time-dependent immune-modulating effect of the sympathetic nervous system conferred by altering the cytokine profile in the local lymph nodes and spleen of mice with type II collagen-induced arthritis. AB - OBJECTIVE: The sympathetic nervous system (SNS) seems to play a proinflammatory role in the early asymptomatic phase of arthritis, but its role in the late stages of chronic arthritis is not well known. The purpose of this study was to examine the effects of the SNS on late-stage chronic arthritis in mice with type II collagen-induced arthritis (CIA). METHODS: We tested the effects of the SNS by ablating sympathetic nerves at different time points in mice with CIA. Early sympathectomy was performed 7 days before immunization. Late sympathectomy was performed on day 56. Cytokine stimulation assays were performed on local lymph node cells and spleen cells, and levels of interleukin-10 (IL-10), IL-4, tumor necrosis factor alpha (TNFalpha), and interferon-gamma (IFNgamma) were determined. RESULTS: Animals with CIA that underwent early sympathectomy showed significantly lower arthritis scores than the controls. In contrast, animals that underwent late sympathectomy had significantly increased arthritis scores compared with controls. On day 0, lymph node cells from animals subjected to early sympathectomy had increased levels of IL-10 and IL-4 and unchanged levels of TNFalpha and IFNgamma compared with those from untreated animals. This indicates an immune-stimulating property of the SNS in draining lymph nodes. On day 80, lymph node cells and spleen cells from animals subjected to late sympathectomy showed increased levels of TNFalpha and IFNgamma compared with those from nonsympathectomized controls with CIA. This indicates an immune depressing property of the SNS in draining lymph nodes and spleen. Arthritis per se largely diminished sympathetic nerve fiber density in synovium on day 80 (P < 0.01). CONCLUSION: The effect of the SNS is bimodal, enhancing or depressing levels of proinflammatory and antiinflammatory cytokines. This feature is dependent on the time point of immune system activation and the respective compartment. The SNS supports inflammation during the asymptomatic phase of CIA, whereas it inhibits inflammation during the chronic symptomatic phase. PMID- 15818684 TI - Pregabalin for the treatment of fibromyalgia syndrome: results of a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: Fibromyalgia syndrome (FMS) is characterized by widespread musculoskeletal pain and lowered pain threshold. Other prominent symptoms include disordered sleep and fatigue. FMS affects an estimated 2% of the population, predominantly women. This trial was designed to evaluate the efficacy and safety of pregabalin, a novel alpha(2)-delta ligand, for treatment of symptoms associated with FMS. METHODS: This multicenter, double-blind, 8-week, randomized clinical trial compared the effects of placebo with those of 150, 300, and 450 mg/day pregabalin on pain, sleep, fatigue, and health-related quality of life in 529 patients with FMS. The primary outcome variable was the comparison of end point mean pain scores, derived from daily diary ratings of pain intensity, between each of the pregabalin treatment groups and the placebo group. RESULTS: Pregabalin at 450 mg/day significantly reduced the average severity of pain in the primary analysis compared with placebo (-0.93 on a 0-10 scale) (P /=50% improvement in pain at the end point (29%, versus 13% in the placebo group; P = 0.003). Pregabalin at 300 and 450 mg/day was associated with significant improvements in sleep quality, fatigue, and global measures of change. Pregabalin at 450 mg/day improved several domains of health-related quality of life. Dizziness and somnolence were the most frequent adverse events. Rates of discontinuation due to adverse events were similar across all 4 treatment groups. CONCLUSION: Pregabalin at 450 mg/day was efficacious for the treatment of FMS, reducing symptoms of pain, disturbed sleep, and fatigue compared with placebo. Pregabalin was well tolerated and improved global measures and health-related quality of life. PMID- 15818683 TI - MRL/Mp CD4+,CD25- T cells show reduced sensitivity to suppression by CD4+,CD25+ regulatory T cells in vitro: a novel defect of T cell regulation in systemic lupus erythematosus. AB - OBJECTIVE: To investigate the hypothesis that loss of suppression mediated by peripheral CD4+,CD25+ regulatory T cells is a hallmark of systemic lupus erythematosus (SLE). METHODS: Mice of the MRL/Mp strain were studied as a polygenic model of SLE. Following immunomagnetic selection, peripheral lymphoid CD25+ and CD25- CD4+ T cells were cultured independently or together in the presence of anti-CD3/CD28 monoclonal antibody-coated beads. Proliferation was assessed by measuring the incorporation of tritiated thymidine. RESULTS: While MRL/Mp CD4+,CD25+ regulatory T cells showed only subtle abnormalities of regulatory function in vitro, syngeneic CD4+,CD25- T cells showed significantly reduced sensitivity to suppression, as determined by crossover experiments in which MRL/Mp CD4+,CD25- T cells were cultured with H-2-matched CBA/Ca CD4+,CD25+ regulatory T cells in the presence of a polyclonal stimulus. CONCLUSION: Our findings highlight a novel defect of peripheral tolerance in SLE. Identification of this defect could open new opportunities for therapeutic intervention. PMID- 15818685 TI - Elevated expression of the Brn-3a and Brn-3b transcription factors in systemic lupus erythematosus correlates with antibodies to Brn-3 and overexpression of Hsp90. AB - OBJECTIVE: Important developmental and antiapoptotic roles have been described for the Brn-3 family of transcription factors in mammalian cells. Following a report of pathogenic autoantibody-inducing T cell reactivity to the Brn-3 transcription factors in murine lupus, we undertook this study to investigate serum levels of antibodies to Brn-3 and levels of expression of Brn-3 in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE). METHODS: Serum and PBMC samples were obtained from 87 SLE patients and 30 normal control subjects. Serum antibodies to the Brn-3a and Brn 3b transcription factors were measured by enzyme-linked immunosorbent assay. Levels of Brn-3a and Brn-3b messenger RNA (mRNA) in PBMCs were measured by reverse transcription and real-time quantitative polymerase chain reaction. RESULTS: Elevated serum levels of antibodies to Brn-3a and Brn-3b were found in 43% and 32%, respectively, of SLE patients. This elevation paralleled enhanced expression of Brn-3a and Brn-3b in PBMCs of 44% and 31%, respectively, of SLE patients. Furthermore, we observed a significant correlation (P = 0.002) between elevated levels of anti-Brn-3b antibodies and elevated levels of Brn-3b mRNA in individual patients. A preliminary analysis of possible target genes for Brn-3a and Brn-3b revealed a significant correlation (P = 0.01) between the level of Brn 3a mRNA and the level of Hsp90 protein (90-kd heat-shock protein, which is overexpressed in SLE) in PBMCs of SLE patients. In addition, we observed that overexpression of Brn-3a and Brn-3b in cultured cells enhanced expression of Hsp90 protein and transcription of Hsp90 promoter-reporter constructs. Finally, we observed an association between elevated levels of Brn-3a mRNA and active SLE (P = 0.002). CONCLUSION: Expression of both Brn-3a and Brn-3b was found to be enhanced in SLE, and this correlated with enhanced levels of autoantibodies to these proteins and with the previously reported overexpression of Hsp90, which was shown to be a novel gene regulated by Brn-3a and Brn-3b. The overexpression of Brn-3a correlated with active disease, suggesting that it may play a role in the disease process via its targeting by the immune system and its ability to induce the expression of specific genes. PMID- 15818686 TI - Expression of interleukin-22 in rheumatoid arthritis: potential role as a proinflammatory cytokine. AB - OBJECTIVE: Interleukin-22 (IL-22) is a novel cytokine of the IL-10 family. Although its pathophysiologic function is largely unknown, induction of acute phase responses by IL-22 has suggested proinflammatory properties. In this study, we sought to examine whether IL-22 plays a role in the pathogenesis of rheumatoid arthritis (RA). METHODS: Expression of IL-22 and IL-22 receptor 1 (IL-22R1) was examined by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunohistochemical analysis. The effects of recombinant IL-22 (rIL-22) on cultured synovial fibroblasts derived from RA patients (RASF), with regard to the proliferation of synovial fibroblasts and production of monocyte chemoattractant protein 1 (MCP-1), were examined by alamer blue assay and enzyme linked immunosorbent assay, respectively. RESULTS: IL-22 messenger RNA was detected by RT-PCR in RA synovial tissues and mononuclear cells isolated from RA synovial fluid samples. High levels of IL-22 were expressed both in the lining and the sublining layers of RA synovial tissues. Staining for vimentin and CD68, as markers of synovial fibroblasts and macrophages, respectively, showed that the majority of IL-22-positive cells were synovial fibroblasts and macrophages. IL 22R1 was also expressed in both the lining and the sublining layers of RA synovial tissues. The majority of cells expressing IL-22R1 were positive for vimentin, but not for CD68. Expression of IL-22 and IL-22R1 in RASF was confirmed by RT-PCR and Western blot analysis. In vitro, rIL-22 significantly increased proliferation of RASF and production of MCP-1 by RASF above the value of medium controls. Moreover, MAPK activation was induced in RASF in response to IL-22 stimulation. CONCLUSION: These data suggest that IL-22, produced by synovial fibroblasts and macrophages, promotes inflammatory responses in RA synovial tissues by inducing the proliferation and chemokine production of synovial fibroblasts. PMID- 15818687 TI - The treatment of rheumatoid arthritis: getting better all the time? PMID- 15818688 TI - Familial aggregation of systemic lupus erythematosus, rheumatoid arthritis, and other autoimmune diseases in 1,177 lupus patients from the GLADEL cohort. AB - OBJECTIVE: To determine whether there is familial aggregation of systemic lupus erythematosus (SLE) and/or other autoimmune diseases in SLE patients and to identify clinical differences between patients with and those without familial autoimmunity. METHODS: We interviewed members of the Grupo Latinoamericano de Estudio del Lupus Eritematoso (GLADEL) inception cohort of 1,214 SLE patients to ascertain whether they had relatives with SLE and/or other autoimmune diseases. Identified relatives were studied. Familial aggregation was tested using reported highest and intermediate population prevalence data for SLE, rheumatoid arthritis (RA), or all autoimmune diseases, and studies were performed to identify the genetic model applicable for SLE. RESULTS: We identified 116 first-, second-, or third-degree relatives with SLE, 79 with RA, 23 with autoimmune thyroiditis, 3 with scleroderma, 1 with polymyositis, and 16 with other autoimmune diseases, related to 166 of the 1,177 SLE patients in the GLADEL cohort who agreed to participate. Forty-two SLE patients had 2 or more relatives with an autoimmune disease. We found a lambda(sibling) of 5.8 and 29.0 for SLE and of 3.2-5.3 for RA, when comparing with their reported high or intermediate population prevalence, respectively. We also found familial aggregation for autoimmune disease in general (lambda(sibling) = 1.5) and determined that for SLE, a polygenic additive genetic model, rather than a multiplicative one, is applicable. CONCLUSION: In SLE there is familial aggregation of SLE, RA, and autoimmune disease in general. A polygenic additive model applies for SLE. American Indian-white Mestizo SLE patients and those with higher socioeconomic level were more likely to have familial autoimmunity. PMID- 15818689 TI - Prophylactic adenovirus-mediated human kallistatin gene therapy suppresses rat arthritis by inhibiting angiogenesis and inflammation. AB - OBJECTIVE: Kallistatin has been shown to be an angiogenesis inhibitor. In this study, we investigated whether adenovirus-mediated kallistatin gene delivery has a prophylactic effect in a rat arthritis model. METHODS: Adenovirus containing the human kallistatin gene (AdHKBP) was injected intraarticularly into ankle joints before the onset of arthritis in a rat model. The effect of kallistatin gene transfer on endothelial cell proliferation in joint extracts was assayed. The response to kallistatin treatment was determined according to clinical parameters, including ankle circumference, articular index, and radiographic scores. Hematoxylin and eosin staining was performed in order to score joint tissues and count neutrophil numbers. In addition, small vessels were quantified by identification of von Willebrand factor-positive endothelial cells. The inflammatory responses were determined by measuring tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta) levels in ankle homogenates. RESULTS: The expression of recombinant human kallistatin in rat ankle joints after gene transfer was identified by immunohistochemical analysis and Western blotting. Significant reductions in the ankle circumference, articular index, and radiographic score were observed in AdHKBP-treated rats compared with control rats treated with the adenoviral plasmid carrying green fluorescent protein. Kallistatin gene transfer also significantly ameliorated the histologic scores in ankle joints and reduced vessel density and neutrophil numbers. The inhibitory effect of kallistatin on the accumulation of inflammatory cells in ankle joints was accompanied by reduced TNFalpha and IL-1beta levels in joint homogenates. Furthermore, an in vitro experiment showed that the proliferation of endothelial cells was markedly inhibited by the addition of AdHKBP-treated joint extract to the culture media, supporting a role of kallistatin in inhibiting angiogenesis. CONCLUSION: This study demonstrates that kallistatin gene therapy has a prophylactic effect in inhibiting arthritis in the rat ankle. Kallistatin inhibits arthritis through its antiangiogenesis and antiinflammation activities. These results implicate potential therapeutic applications for suppression of arthritis by kallistatin gene therapy. PMID- 15818690 TI - Development of fatal tuberculosis in a patient with rheumatoid arthritis after three years of treatment with infliximab: comment on the article by Wolfe et al. PMID- 15818691 TI - Risk factors for functional decline in older adults with arthritis. AB - OBJECTIVE: Functional limitation is a major factor in medical costs. This study was undertaken to evaluate the prevalence of functional limitation among adults with arthritis and the frequency of functional decline over 2 years, and to investigate factors amenable to public health intervention that predict functional decline. METHODS: Longitudinal data (1998-2000) from a cohort of 5,715 adults ages 65 years or older with arthritis from a national probability sample were analyzed. Function was defined based on ability to perform basic activities of daily living (ADL) tasks and instrumental ADL. Adjusted odds ratios (ORs) from a multiple logistic regression model were used to estimate the associations between functional decline and comorbid conditions, health behaviors, and economic factors. RESULTS: Overall, 19.7% of this cohort had functional limitation at baseline, including 12.9% with ADL limitations. Over the subsequent 2 years, function declined in 13.6% of those at risk. Functional decline was most frequent among women (15.0%) and minorities (18.0% Hispanics, 18.7% African Americans) with arthritis. Lack of regular vigorous physical activity, the most prevalent risk factor (64%), almost doubled the odds of functional decline (adjusted OR 1.9, 95% confidence interval 1.5-2.4) after controlling for all risk factors. It was found that if all subjects engaged in regular vigorous physical activity, the expected functional decline could be reduced as much as 32%. Other significant predictors included older age, cognitive impairment, depressive symptoms, diabetes, physical limitations, no alcohol use, stroke, and vision impairment. CONCLUSION: Lack of regular vigorous physical activity is a potentially modifiable risk factor that could substantially reduce functional decline and associated health care costs. Prevention/intervention programs should include regular vigorous physical activity, weight maintenance, and medical intervention for health needs. (c) 2005, American College of Rheumatology. PMID- 15818692 TI - Reduced tumor necrosis factor signaling in primary human fibroblasts containing a tumor necrosis factor receptor superfamily 1A mutant. AB - OBJECTIVE: Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is an autoinflammatory syndrome associated with mutations in the gene that encodes tumor necrosis factor receptor superfamily 1A (TNFRSF1A). The purpose of this study was to describe a novel TNFRSF1A mutation (C43S) in a patient with TRAPS and to examine the effects of this TNFRSF1A mutation on tumor necrosis factor alpha (TNFalpha)-induced signaling in a patient-derived primary dermal fibroblast line. METHODS: TNFRSF1A shedding from neutrophils was measured by flow cytometry and enzyme-linked immunosorbent assay (ELISA). Primary dermal fibroblast lines were established from the patient with the C43S TRAPS mutation and from healthy volunteers. Activation of NF-kappaB and activator protein 1 (AP-1) was evaluated by electrophoretic mobility shift assays. Cytokine production was measured by ELISA. Cell viability was measured by alamar blue assay. Apoptosis was measured by caspase 3 assay in the fibroblasts and by annexin V assay in peripheral blood mononuclear cells. RESULTS: Activation-induced shedding of the TNFRSF1A from neutrophils was not altered by the C43S TRAPS mutation. TNFalpha-induced activation of NF-kappaB and AP-1 was decreased in the primary dermal fibroblasts with the C43S TNFRSF1A mutation. Nevertheless, the C43S TRAPS fibroblasts were capable of producing interleukin-6 (IL-6) and IL-8 in response to TNFalpha. However, TNFalpha-induced cell death and apoptosis were significantly decreased in the samples from the patient with the C43S TRAPS mutation. CONCLUSION: The C43S TNFRSF1A mutation results in decreased TNFalpha-induced nuclear signaling and apoptosis. Our data suggest a new hypothesis, in that the C43S TRAPS mutation may cause the inflammatory phenotype by increasing resistance to TNFalpha-induced apoptosis. PMID- 15818693 TI - Effect of oral glucosamine on cartilage degradation in a rabbit model of osteoarthritis. AB - OBJECTIVE: To determine whether oral glucosamine alleviates cartilage degradation in an animal model of osteoarthritis (OA). METHODS: The effect of 8 weeks of daily oral glucosamine hydrochloride on degeneration of articular cartilage was evaluated in rabbits in which anterior cruciate ligament transection (ACLT) was performed to induce OA. Animals were treated with glucosamine (n = 16) or a placebo (n = 16) and necropsied at 11 weeks. Seven unoperated rabbits served as controls. The articular cartilage was evaluated macroscopically and histologically and analyzed for total type II collagen and glycosaminoglycan (GAG) content. RESULTS: Histologic analysis revealed that loss of proteoglycan, based on Safranin O-fast green staining, was significantly reduced in the lateral tibial plateau cartilage of ACL-transected limbs in the glucosamine group compared with ACL-transected limbs in the placebo group, with a similar, but not significant, trend for the lateral femoral condylar cartilage. Likewise, macroscopic analysis of cartilage showed that the lateral tibial plateau alone had a significantly lower rate of disease in the glucosamine group, which was consistent with the results of the independent histologic assessment. However, no significant treatment effect was detected when composite histologic scores were analyzed. A significant reduction in GAG content was observed in the femoral condyles of placebo-treated ACL-transected joints, but not in the same region of glucosamine-treated ACL-transected joints, compared with their respective contralateral unoperated joints. CONCLUSION: Oral administration of glucosamine had a detectable, site-specific, partial disease-modifying effect in this model of OA. From a clinical perspective, the administration of glucosamine did not prevent fibrillation and/or erosions of the articular cartilage in all of the treated animals, and no effects were detected in the medial joint compartments. PMID- 15818694 TI - Magnetic resonance imaging examinations of the spine in patients with ankylosing spondylitis before and after therapy with the tumor necrosis factor alpha receptor fusion protein etanercept. AB - OBJECTIVE: To assess spinal inflammation by magnetic resonance imaging (MRI) before and after treatment with the tumor necrosis factor receptor fusion protein etanercept compared with placebo. METHODS: As part of a recently published randomized, controlled trial, 40 patients with ankylosing spondylitis (AS) underwent MRI of the lower thoracic and lumbar spine at 4 different time points: baseline, 12 weeks, 24 weeks, and 48 weeks. Nineteen patients received subcutaneous etanercept twice weekly (25 mg twice daily) for 1 year, and 21 patients received placebo for 6 months before being switched to etanercept. The mean age of the patients was 39.7 years, 75% were male, 89% were HLA-B27 positive, and the mean disease duration was 13 years. MRI examinations included T1-weighted sequences before and after application of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) and T2-weighted fat-saturated sequences. MRI sequences were scored with an adjusted AS spinal MRI score, using predefined vertebral units (VUs) as a basis. RESULTS: After 12 weeks, spinal inflammation (as assessed by T2-weighted MRI with fat saturation) regressed by 54% in the etanercept group (mean score 1.33 per VU at baseline and 0.61 per VU at 12 weeks; P = 0.002) but worsened by 13% in the placebo group (0.94 at baseline and 1.06 at 12 weeks) (P < 0.001 between groups). After switching to etanercept, placebo patients improved similarly. T1-weighted Gd-DTPA MRI sequences performed equally well in detecting spinal changes. At baseline, >50% of all active lesions were detected in the thoracic spine. Deterioration of chronic changes was significant only in patients treated with placebo. CONCLUSION: Etanercept treatment of patients with active AS results in regression of spinal inflammation as assessed by spinal MRI. Inclusion of the thoracic spine in MRI examinations of patients with AS may be of particular importance. PMID- 15818695 TI - Factors associated with hip cartilage volume measured by magnetic resonance imaging: the Tasmanian Older Adult Cohort Study. AB - OBJECTIVE: To compare associations between anthropometric and lifestyle factors and femoral head cartilage volume/thickness and radiographic features of osteoarthritis (OA) and to provide evidence of construct validity for magnetic resonance imaging (MRI) assessment of femoral cartilage volume and thickness. METHODS: We studied a cross-sectional sample of 151 randomly selected subjects (79 men, 72 women; mean age 63 years) from the Tasmanian Older Adult Cohort Study. A sagittal T1-weighted fat-suppression MRI scan of the right hip was performed to determine femoral head cartilage volume, cartilage thickness, and size. An anteroposterior radiograph of the pelvis with weight bearing was performed and scored for radiographic evidence of OA in the right hip. Other factors measured were height, weight, leg strength, serum vitamin D levels, and bone mineral density. RESULTS: Hip cartilage volume was significantly associated with female sex, body mass index, and femoral head size, whereas hip cartilage thickness was significantly associated only with the size of the femoral head. Only female sex was significantly associated with the total radiographic OA score and the joint space narrowing (JSN) score, but not the osteophyte score. Radiographic JSN of the hip, especially axial JSN (but not osteophytes), was significantly correlated with hip cartilage volume and thickness. CONCLUSION: Femoral head cartilage volume and thickness have modest but significant construct validity when correlated with radiographic findings. Furthermore, the generally stronger associations with volume compared with radiographic OA suggest that MRI may be superior at identifying risk factors for hip OA. PMID- 15818696 TI - Inhibition of integrative cartilage repair by proteoglycan 4 in synovial fluid. AB - OBJECTIVE: To determine the effects of the articular cartilage surface, as well as synovial fluid (SF) and its components, specifically proteoglycan 4 (PRG4) and hyaluronic acid (HA), on integrative cartilage repair in vitro. METHODS: Blocks of calf articular cartilage were harvested, some with the articular surface intact and others without. Some of the latter types of blocks were pretreated with trypsin, and then with bovine serum albumin, SF, PRG4, or HA. Immunolocalization of PRG4 on cartilage surfaces was performed after treatment. Pairs of similarly treated cartilage blocks were incubated in partial apposition for 2 weeks in medium supplemented with serum and (3)H-proline. Following culture, mechanical integration between apposed cartilage blocks was assessed by measuring adhesive strength, and protein biosynthesis and deposition were determined by incorporated (3)H-proline. RESULTS: Samples with articular surfaces in apposition exhibited little integrative repair compared with samples with cut surfaces in apposition. PRG4 was immunolocalized at the articular cartilage surface, but not in deeper, cut surfaces (without treatment). Cartilage samples treated with trypsin and then with SF or PRG4 exhibited an inhibition of integrative repair and positive immunostaining for PRG4 at treated surfaces compared with normal cut cartilage samples, while samples treated with HA exhibited neither inhibited integrative repair nor PRG4 at the tissue surfaces. Deposition of newly synthesized protein was relatively similar under conditions in which integration differed significantly. CONCLUSION: These results support the concept that PRG4 in SF, which normally contributes to cartilage lubrication, can inhibit integrative cartilage repair. This has the desirable effect of preventing fusion of apposing surfaces of articulating cartilage, but has the undesirable effect of inhibiting integrative repair. PMID- 15818697 TI - Evidence of radiographic benefit of treatment with infliximab plus methotrexate in rheumatoid arthritis patients who had no clinical improvement: a detailed subanalysis of data from the anti-tumor necrosis factor trial in rheumatoid arthritis with concomitant therapy study. AB - OBJECTIVE: To assess the relationship between inflammation and joint destruction in rheumatoid arthritis (RA) patients who have not responded clinically to treatment. METHODS: Changes from baseline to week 54 in clinical variables and measures of radiographic progression were compared between patients who received infliximab (3 mg/kg or 10 mg/kg every 4 or 8 weeks) plus methotrexate (MTX) and those who received MTX plus placebo in the Anti-Tumor Necrosis Factor Trial in RA with Concomitant Therapy trial. RESULTS: At week 54, patients who did not show 20% improvement by American College of Rheumatology criteria (ACR20 nonresponders) while receiving infliximab plus MTX exhibited mild but statistically significant improvement in clinical variables, including the 28 joint Disease Activity Score (DAS28) (P < 0.001), tender joint count (P = 0.014), swollen joint count (P < 0.001), and C-reactive protein (CRP) level (P < 0.001). Whereas the clinical and CRP changes among ACR20 nonresponders to infliximab plus MTX were small and much lower than among ACR20 responders to this treatment, radiographic progression among ACR20 nonresponders to infliximab plus MTX was significantly inhibited (P < 0.001) compared with ACR20 nonresponders to MTX plus placebo. Radiographic progression was much greater in patients receiving MTX plus placebo than in patients receiving infliximab plus MTX, irrespective of ACR response status (mean change in modified Sharp/van der Heijde score 6.0 in ACR20 responders and 7.2 in ACR20 nonresponders in the MTX plus placebo-treated group, versus 0.1 in ACR20 responders and 1.2 in ACR20 nonresponders in the infliximab plus MTX-treated group). Furthermore, among patients who were ACR20 nonresponders through week 54, patients who were DAS nonresponders at weeks 30 and 54, and patients without any improvement in individual clinical variables, those receiving infliximab plus MTX still demonstrated inhibition of structural damage that was statistically significant compared with inhibition in patients who received MTX plus placebo (P < 0.05 to P < 0.001). CONCLUSION: Even in patients without clinical improvement, treatment with infliximab plus MTX provided significant benefit with regard to the destructive process, suggesting that in such patients these 2 measures of disease are dissociated. PMID- 15818698 TI - Continuous indices of core data set measures in rheumatoid arthritis clinical trials: lower responses to placebo than seen with categorical responses with the American College of Rheumatology 20% criteria. AB - OBJECTIVE: To describe indices that are continuous counterparts of categorical responses to the American College of Rheumatology 20% improvement criteria (ACR20), ACR50, and ACR70, which extend rheumatoid arthritis (RA) clinical trial results and recognize clinical worsening (as well as improvement) with active and placebo treatments. METHODS: Data from a clinical trial of leflunomide, methotrexate, and placebo treatment over 1 year were reanalyzed. Percent change was computed for each of the 7 components of the ACR core set of outcome measures. Four continuous indices were computed: 1) ACR-N (lowest of 3 values: number of swollen joints, number of tender joints, and median of the other 5 measures); 2) composite (median of all 7 measures [3 patient and 3 assessor measures plus erythrocyte sedimentation rate]); 3) patient-only (median of physical function, pain, and global status); and 4) assessor-only (median of number of swollen joints, number of tender joints, and global status). Means, medians, categorical 20%, 50%, and 70% responses, and continuous probability plots were computed according to each index for the 3 treatment groups and were compared with one another and with standard ACR20, ACR50, and ACR70 responses. RESULTS: Mean levels of improvement calculated using the different methods, in patients taking leflunomide, placebo, and methotrexate, respectively, were as follows: ACR-N 20%, -12%, and 13%; composite 43%, 9%, and 33%; patient-only 36%, 0%, and 26%; assessor-only 50%, 20%, and 44%; and ACR20 52%, 26%, and 46%. Differences between leflunomide and placebo were 30-36%, and differences between methotrexate and placebo were 24-26%. CONCLUSION: Continuous indices may be an informative addition to categorical ACR 20%, 50%, or 70% responses to compare efficacies of various treatments in RA, and to describe lower responses to placebo by recognizing worsening as well as improvement. PMID- 15818699 TI - Sustained benefits of infliximab therapy for dermatologic and articular manifestations of psoriatic arthritis: results from the infliximab multinational psoriatic arthritis controlled trial (IMPACT). AB - OBJECTIVE: To investigate the efficacy and tolerability of infliximab therapy for the articular and dermatologic manifestations of active psoriatic arthritis (PsA). METHODS: One hundred four patients with PsA in whom prior therapy with at least 1 disease-modifying antirheumatic drug (DMARD) had failed were recruited into this investigator-initiated, multicenter, randomized, double-blind, placebo controlled clinical trial. During the initial blinded portion of the study, patients received infusions of infliximab (5 mg/kg) or placebo at weeks 0, 2, 6, and 14. After week 16, patients initially assigned to receive placebo crossed over to receive infliximab 5 mg/kg every 8 weeks through week 50, while patients initially randomized to infliximab continued to receive active treatment at the same dose through week 50. The primary efficacy outcome was achievement of the American College of Rheumatology 20% criteria for improvement in rheumatoid arthritis (ACR20) at week 16. Additional predefined clinical efficacy assessments included the Psoriasis Area and Severity Index (PASI) score, the ACR50 and ACR70 criteria, the Disease Activity Score in 28 joints, the Health Assessment Questionnaire, ratings of enthesitis and dactylitis, and the Psoriatic Arthritis Response Criteria score. RESULTS: The proportion of infliximab-treated patients who achieved an ACR20 response at week 16 (65%) was significantly higher than the proportion of placebo-treated patients who achieved this response (10%). In addition, 46% of infliximab-treated patients achieved an ACR50 response, and 29% achieved an ACR70 response; no placebo-treated patient achieved these end points. Among patients who had PASI scores of >/=2.5 at baseline, 68% of infliximab treated patients achieved improvement of >/=75% in the PASI score at week 16 compared with none of the placebo-treated patients. Continued therapy with infliximab resulted in sustained improvement in articular and dermatologic manifestations of PsA through week 50. The incidence of adverse events was similar between the treatment groups. CONCLUSION: Therapy with infliximab at a dose of 5 mg/kg significantly improved the signs and symptoms of arthritis, psoriasis, dactylitis, and enthesitis in patients with active PsA that had been resistant to DMARD therapy. With continued infliximab treatment, benefits were sustained through 50 weeks. The benefit-to-risk ratio appeared favorable in this study population. PMID- 15818701 TI - Generation of novel covalent RNA-protein complexes in cells by ultraviolet B irradiation: implications for autoimmunity. AB - OBJECTIVE: To determine whether ultraviolet B (UVB) irradiation induces novel modifications in autoantigens targeted during experimental photoinduced epidermal damage. METHODS: To search for novel UVB-induced autoantigen modifications, lysates made from UVB-irradiated human keratinocytes or HeLa cells were immunoblotted using human autoantibodies that recognize ribonucleoprotein autoantigens. Novel autoantigen structures identified were further characterized using nucleases and RNA hybridization. RESULTS: Human sera that recognize U1-70 kd (U1-70K) and La by immunoblotting also recognized multiple novel species when they were used to immunoblot lysates of UVB-irradiated keratinocytes or HeLa cells. These species were not present in control cells and were not observed when apoptosis was induced by Fas ligation or cytotoxic lymphocyte granule contents. Biochemical analysis using multiple assays revealed that these novel UVB-induced molecular species result from the covalent crosslinking between the U1 RNA and the hYRNA molecules with their associated proteins, including U1-70K, La, and likely components of the Sm particle. CONCLUSION: These data demonstrate that UVB irradiation of live cells can directly induce covalent RNA-protein complexes, which are recognized by human autoantibodies. As previously described for other autoantigens, these covalent complexes of RNA and proteins may have important consequences in terms of antigen capture and processing. PMID- 15818702 TI - Evaluation of the efficacy of etoricoxib in ankylosing spondylitis: results of a fifty-two-week, randomized, controlled study. AB - OBJECTIVE: To assess the efficacy, safety, and tolerability of etoricoxib, a cyclooxygenase 2 (COX-2) selective inhibitor, administered continuously over 52 weeks for the treatment of ankylosing spondylitis (AS). METHODS: This 2-part, multicenter, double-blind, parallel-group, 52-week study evaluated 2 doses of etoricoxib (90 and 120 mg) compared with naproxen at 1,000 mg. A 6-week, active comparator- and placebo-controlled period (part I) was followed by a 46-week active-comparator-controlled period (part II). The primary outcome measures (on 100-mm visual analog scales) were patient's assessment of spine pain, patient's global assessment of disease activity, and the Bath Ankylosing Spondylitis Functional Index. RESULTS: Of the 387 patients randomized to receive treatment, 301 (77.8%) completed part I and 284 (75.9%) completed part II. Compared with placebo over 6 weeks, those receiving 90 mg etoricoxib, 120 mg etoricoxib, and naproxen demonstrated significantly (P < 0.001) greater improvement in all primary end points; treatment effects (expressed as the difference in least squares mean change versus placebo) were 21-29 mm for spine pain, 18-25 mm for disease activity, and 11-15 mm for function. Compared with patients receiving naproxen, significantly greater improvement in all primary end points was demonstrated in the combined group receiving either 90 mg etoricoxib or 120 mg etoricoxib over 6 weeks, in each individual etoricoxib treatment group over 6 weeks, and in the combined etoricoxib group over 1 year (all P < 0.05); results for secondary and exploratory end points were generally consistent with those from the primary analysis. Among all groups, there were no significant differences in the incidence of overall clinical, drug-related, or serious adverse experiences (AEs) and discontinuations due to AEs. Safety observations during part II were generally consistent with those in part I. CONCLUSION: Etoricoxib at doses of 90 mg and 120 mg demonstrated superior efficacy compared with placebo over 6 weeks, and compared with naproxen over 1 year. These study results demonstrate that etoricoxib is generally safe, well-tolerated, and efficacious for the treatment of AS. PMID- 15818703 TI - Urinary type II collagen helical peptide (HELIX-II) as a new biochemical marker of cartilage degradation in patients with osteoarthritis and rheumatoid arthritis. AB - OBJECTIVE: Type II collagen, which consists of a large helical domain and telopeptides at each end, is the most abundant protein of cartilage matrix. The aim of this study was to develop a biochemical marker reflecting the degradation of the helical region of type II collagen and to evaluate its clinical performance in patients with osteoarthritis (OA) and rheumatoid arthritis (RA). METHODS: We developed a competitive polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) using the 622-632 peptide derived from the sequence of the alpha1 chain of human type II collagen (HELIX-II) as immunogen and standard. We measured urinary levels of HELIX-II peptide and C-terminal crosslinking telopeptide of type II collagen (CTX-II) in 90 patients with knee OA (73% women; mean +/- SD age 63.0 +/- 8.0 years, mean +/- SD disease duration 6.1 +/- 6.8 years), 89 patients with early RA (disease duration /=0.5 units/year), with an odds ratio (OR) of 5.9 (95% confidence interval [95% CI] 2.0-17.2) after adjustment for serum C-reactive protein (CRP) levels and baseline joint damage. Patients with increased levels of both urinary HELIX-II and CTX-II had the highest risk of progression (OR 17.5 [95% CI 3.1-99]). CONCLUSION: The HELIX-II ELISA is specific for type II collagen degradation, has adequate technical performance, and can distinguish patients with knee OA or RA from healthy controls. Elevated HELIX-II levels are associated with increased risk of radiographic progression in RA independently of CRP levels, baseline joint damage, and urinary CTX-II levels. The HELIX-II ELISA should be useful for the clinical investigation of patients with arthritis and for identifying RA patients at higher risk of progression. PMID- 15818704 TI - Homeostatic effects of the metalloproteinase disintegrin ADAM15 in degenerative cartilage remodeling. AB - OBJECTIVE: The membrane-anchored metalloproteinase disintegrin ADAM15 is up regulated in osteoarthritis and has been implicated in proteolysis and cell matrix interactions. To address its role in cartilage metabolism, we performed an analysis of joint morphology in aging mice with a targeted inactivation of the ADAM15 gene (ADAM15(-/-)). In addition, a human chondrocyte cell line overexpressing ADAM15 was used to investigate the role of ADAM15 in an in vitro model of chondrocyte-matrix interactions. METHODS: Knee joint sections from 3-, 6 , and 12-14-month-old ADAM15(-/-) and wild-type (WT) 129/SvJ mice were examined for synovial hyperplasia, cartilage degradation, and osteophyte formation. Stable transfection of the human T/C28a4 chondrocyte cell line with full-length human ADAM15 complementary DNA led to the establishment of ADAM15-overexpressing chondrocytes that were further analyzed for their capability to adhere to and to survive on cartilage matrix molecules (fibronectin and types II and VI collagen) under conditions of serum starvation. ADAM15 expression was investigated by reverse transcription-polymerase chain reaction and Western blotting. RESULTS: Aging ADAM15(-/-) mice exhibited accelerated development of osteoarthritic lesions compared with WT mice, and the difference was statistically significant at age 12 months. The osteoarthritic changes preferentially affected male ADAM15( /-) mice. ADAM15 overexpression in T/C28a4 cells led to the specific reinforcement of chondrocyte adhesion to cartilage types II and VI collagen, and this was associated with enhanced cell viability under conditions of serum starvation. CONCLUSION: The accelerated development of murine osteoarthritis in ADAM15 deficiency as well as the proadhesive and cell survival-promoting in vitro effect of ADAM15 overexpression suggest a homeostatic rather than a destructive role of ADAM15 in cartilage remodeling. PMID- 15818705 TI - Identifying an early marker for congenital heart block: when is a long PR interval too long? Comment on the article by Sonesson et al. PMID- 15818706 TI - Patients seen for standard rheumatoid arthritis care have significantly better articular, radiographic, laboratory, and functional status in 2000 than in 1985. AB - OBJECTIVE: A comprehensive quantitative analysis of measures of disease activity and joint damage has not been available to compare patients in different eras in the same clinical setting. This study was undertaken to determine whether the clinical status of patients with rheumatoid arthritis (RA) has improved on average in recent years. METHODS: A quantitative cross-sectional evaluation, which included joint count, radiographic, laboratory, patient questionnaire, and physical function measures, and therapies, was performed in 125 consecutive RA patients seen from 1984 through 1986 ("1985 cohort"). A virtually identical assessment was performed in 150 patients seen from 1999 through 2001 ("2000 cohort"), in the same weekly academic clinic. Measures were compared using descriptive statistics and a median regression model, adjusted for age, duration of disease, level of formal education, and rheumatoid factor. RESULTS: Patients in 1985 had significantly poorer status compared with those in 2000: median 12 versus 5 swollen joints, Larsen radiographic score 20 versus 3, erythrocyte sedimentation rate 33 mm/hour versus 20, and modified Health Assessment Questionnaire 1.0 versus 0.4 (P < 0.019). Severe Disease Activity Scores >5.1 were seen in 69% of 1985 patients, compared with 30% in 2000. Methotrexate was taken by 10% of patients in 1985, versus 76% in 2000. The proportion of patients not taking any disease-modifying antirheumatic drugs was 66% in 1985 versus 13% in 2000. CONCLUSION: Patients receiving standard care for RA in this setting had significantly better status, including radiographic scores, in 2000 than in 1985, associated with aggressive treatment strategies, prior to the introduction of biologic agents. PMID- 15818707 TI - Interleukin-1 blockade by anakinra improves clinical symptoms in patients with neonatal-onset multisystem inflammatory disease. AB - Neonatal-onset multisystem inflammatory disease (NOMID) is a rare, childhood onset disease that is characterized by chronic, systemic inflammation. The purpose of this report is to describe the effects of interleukin-1 (IL-1) blockade on the clinical symptoms of 2 patients with NOMID. At the time of this report, the patients had been treated with anakinra (Kineret), a recombinant human IL-1 receptor antagonist, for 1.5 and 2 years, respectively. Both patients demonstrated rapid improvement in clinical symptoms and laboratory markers of inflammation. The use of anakinra in these patients seemed to be effective, without any safety concerns. These observations suggest that IL-1 plays a critical role in the pathogenesis of this inflammatory disease, and that blockade with anakinra should be further studied as a treatment for patients with NOMID and related disorders. PMID- 15818708 TI - Is cigarette smoking a risk factor for systemic lupus erythematosus? Comment on the article by Costenbader et al. PMID- 15818709 TI - Successful treatment with intraarticular infliximab for resistant knee monarthritis in a patient with spondylarthropathy: a role for scintigraphy with 99mTc-infliximab. AB - Positive experiences with intraarticular infliximab have been reported in patients with rheumatoid arthritis, ankylosing spondylitis, and Behcet's disease. We used intraarticular infliximab to treat resistant knee monarthritis in a patient with spondylarthropathy. Clinical and laboratory improvement was associated with improvement in scintigraphic findings. This approach is less expensive than intravenous administration of infliximab. We suggest that selection of candidates for this innovative therapy should be guided by anti tumor necrosis factor alpha scintigraphy. PMID- 15818710 TI - The incidence and natural history of Raynaud's phenomenon in the community. AB - OBJECTIVE: Raynaud's phenomenon (RP) is a common disorder, yet its incidence and natural history are unknown. Our objective was to determine the incidence and natural history of RP not associated with a connective tissue disease in a large, community-based population. METHODS: Using serial examinations of the Framingham Heart Study offspring cohort, we collected data regarding RP symptoms for 717 women and 641 men over a 7-year period. We used validated criteria for RP classification and categorized participants as having incident, persistent, or remitted RP. We performed sex-specific analyses of RP status by age, body mass index, vibratory tool use, season of examination, state of residence, use of antihypertensive medications, and smoking status. RESULTS: The mean +/- SD age of participants was 53.5 +/- 10 years. The incidence of RP was 2.2% in women (n = 14) and 1.5% in men (n = 9). Of the 78 women and 50 men who had RP at baseline, 36% of women (n = 28) and 36% of men (n = 18) had persistent RP. RP remitted in 64% of women (n = 50) and 64% of men (n = 32), with 41 women and 25 men meeting no or only 1 RP criterion at followup. RP episodes were infrequent and rarely interfered with daily activities. CONCLUSION: This is the first prospective study to determine the incidence and natural history of RP in a community-based cohort. Our data demonstrate that RP not associated with a connective tissue disease is frequently a transient phenomenon and rarely interferes with daily activities. PMID- 15818711 TI - Relationship between anti-double-stranded DNA antibodies and exacerbation of renal disease in patients with systemic lupus erythematosus. AB - OBJECTIVE: To examine the relationship between changes in anti-double-stranded DNA (anti-dsDNA) antibody levels and the risk of renal flare in patients with systemic lupus erythematosus (SLE), using data from 2 randomized, controlled trials. METHODS: Analyses were based on 487 patients with SLE and a history of lupus nephritis who had an anti-dsDNA antibody titer >/=15 IU/ml at baseline, as measured by Farr assay. Results are presented for the combined population of patients, the placebo arms, and the drug treatment arms in which a dsDNA-based bioconjugate (abetimus sodium; LJP 394) was used. RESULTS: Changes in anti-dsDNA antibody levels were inversely correlated with changes in the C3 level (P < 0.0001 in both trials). Cox proportional hazards regression models showed that changes in anti-dsDNA antibody levels correlated with the risk of renal flare. The models predicted that a point estimate of a 50% reduction in anti-dsDNA antibody levels is associated with a 52% reduction (95% confidence interval [95% CI] 26-68%, nominal P = 0.0007) and a 53% reduction (95% CI 33-69%, nominal P < 0.0001) in the risk of renal flare in the 2 trials, respectively. In the 2 trials, the incidence of renal flare was lower in patients with sustained reductions in anti-dsDNA antibodies (3.0% and 4.1%, respectively) than in patients with stable or increasing antibody levels (21.3% and 20.3%, respectively). CONCLUSION: Changes in anti-dsDNA antibody levels were directly correlated with the risk of renal flare and inversely correlated with changes in the C3 level. Reducing anti-dsDNA antibody levels may represent a therapeutic objective in SLE patients with lupus nephritis, because it is associated with a reduced risk of renal flare. PMID- 15818712 TI - Association of Epstein-Barr virus with systemic lupus erythematosus: effect modification by race, age, and cytotoxic T lymphocyte-associated antigen 4 genotype. AB - OBJECTIVE: Epstein-Barr virus (EBV) is hypothesized to play a role in the development of systemic lupus erythematosus (SLE). Cytotoxic T lymphocyte associated antigen 4 (CTLA-4) is important in regulating T cell-mediated immunity, encompassing the first line of response to viral infections, and genetic variation in CTLA-4 has been associated with SLE. This study examined the seroprevalence of EBV in a population-based study of SLE patients from the southeastern United States, and potential interactions with CTLA-4 polymorphisms were assessed. METHODS: Cases comprised 230 subjects recently diagnosed as having SLE (144 African American and 86 white) from university and community-based clinics, and controls comprised 276 age-, sex-, and state-matched subjects (72 African American and 204 white) recruited from driver's license registries. Antibodies to EBV capsid antigen were determined by enzyme-linked immunosorbent assay, with results expressed as positive or negative using the international standardized ratio (ISR) (a ratio of the sample absorbance to a known standard). CTLA-4 genotypes were identified by polymerase chain reaction-based methods. RESULTS: In African Americans, EBV-IgA seroprevalence was strongly associated with SLE (odds ratio [OR] 5.6, 95% confidence interval [95% CI] 3.0-10.6). In whites, the modest association of SLE with EBV-IgA (OR 1.6) was modified by age, in that the strongest association was observed in those older than age 50 years (OR 4.1, 95% CI 1.6-10.4). The seroprevalence of EBV-IgM and that of EBV-IgG were not associated with SLE. Higher EBV-IgG absorbance ratios were observed in SLE patients, with a significant dose response across units of the ISR in African Americans (P < 0.0001). Allelic variation in the CTLA-4 gene promoter (-1661A/G) significantly modified the association between SLE and EBV-IgA (P = 0.03), with a stronger association among those with the -1661AA genotype. CONCLUSION: These findings suggest that repeated or reactivated EBV infection, which results in increased EBV-IgA seroprevalence and higher IgG antibody titers, may be associated with SLE, and that the CTLA-4 genotype influences immune responsiveness to EBV in SLE patients. The observed patterns of effect modification by race, age, and CTLA-4 genotype should be examined in other studies and may help frame new hypotheses regarding the role of EBV in SLE etiology. PMID- 15818713 TI - Prevention of early renal disease, dyslipidaemia and lipid peroxidation in STZ diabetic rats by LR-9 and LR-74, novel AGE inhibitors. AB - BACKGROUND: Increased formation of advanced glycation/lipoxidation endproducts (AGEs/ALEs) has been implicated in the pathogenesis of various diabetic complications. Several compounds have been developed as inhibitors of AGE/ALE formation. We examined the effects of two new AGE/ALE inhibitors, LR-9 and LR-74, on the development of early renal disease and lipid metabolism in streptozotocin (STZ)-induced diabetic rats. METHODS: Diabetic Sprague-Dawley rats were treated with either of the LR compounds for 32 weeks. Progression of renal disease was evaluated by measurements of urinary albumin and plasma creatinine concentrations. AGE/ALE and nitrotyrosine levels in kidneys were determined by immunohistochemistry. AGE-induced chemical modification of the tail tendon collagen and levels of Nepsilon-(carboxymethyl) and (carboxyethyl)- lysines (CML and CEL) in skin collagen were measured. Plasma lipids and their lipid hydroperoxide concentrations were also determined. In vitro, both compounds were tested for inhibiting lipid peroxidation reactions. RESULTS: Treatment of either LR compounds significantly inhibited the increase in albuminuria, creatinaemia, hyperlipidaemia and lipid peroxidation in diabetic rats without any effect on hyperglycaemia. Both compounds also reduced CML-AGE and nitrotyrosine accumulation in kidney glomeruli and tubules, AGE-linked fluorescence and cross linking of tail collagen, and levels of CML and CEL in skin collagen. In vitro, LR compounds inhibited the oxidation of human low-density lipoprotein (LDL). CONCLUSION: Both compounds can inhibit the progression of renal disease and also prevent dyslipidaemia in type-1 diabetic animals. These compounds may have an additional beneficial effect as an antioxidant against lipid peroxidation, and thus may provide alternative therapeutic options for the treatment of various diabetic macrovascular complications. PMID- 15818714 TI - Insulin therapy in pregnancy complicated by diabetes: are insulin analogs a new tool? AB - It has been demonstrated that good metabolic control maintained throughout pregnancy can reduce maternal and fetal complications in diabetes. To achieve good metabolic control, before conception and throughout pregnancy, insulin therapy needs to be optimized, and, in this context, the new insulins currently on the market may help. We therefore review here what is known about the potential benefits and risks related to the use of these new insulins in pregnancy. Clinical and experimental data on insulin lispro strongly suggest that lispro does not have adverse maternal or fetal effects during pregnancy in women with preexisting diabetes, and also that its use in these women results in improved glycemic control, fewer hypoglycemic episodes, and improved patient satisfaction. In women with gestational diabetes mellitus (GDM), the use of insulin lispro is efficient in reducing postprandial hyperglycemia and some neonatal features related to hyperglycemia, thus stressing its usefulness in this condition. As for insulin aspart, clinical data on GDM patients shows the same efficacy as insulin lispro in lowering postprandial hyperglycemia, indicating that insulin aspart may be used in GDM when this condition is characterized by postprandial hyperglycemia. The results of a multicentric study now in progress on the efficacy and safety of insulin aspart in type 1 pregnant diabetic patients will definitely be useful in establishing whether this insulin is safe in pregnancy. For the moment, the use of insulin glargine during pregnancy is not recommended owing to the lack of data on maternal and fetal effects. PMID- 15818715 TI - Need for information and for involvement in decision making among patients with rheumatoid arthritis: a questionnaire survey. AB - OBJECTIVE: To measure the need for information about rheumatoid arthritis (RA) and the level of desire for involvement in treatment decisions among patients with RA. To examine the relationship between these preferences and what factors (sociodemographic, disease, treatment, level of disability, and level of knowledge about RA) associate with these preferences. METHODS: Questionnaire surveys were mailed to a randomly selected group of 600 patients with RA. Need for information and desire for involvement in decision making were measured using a validated tool (the Autonomy Preference Index). RESULTS: The response rate was 57.3%. The need for information was very high. Information seeking preference scores (median 82.5, interquartile range 80.0-92.5) were significantly higher (P < 0.001) than decision making preference scores (mean +/- SD 56.4 +/- 13.6). Need for information and for decision making were both higher in women than men, and associations with these needs differed in men and women. However, younger age and greater knowledge of RA predicted greater need for decision making. There was no correlation between need for information and for involvement in treatment decisions for either sex (women: r(s) = 0.09, P = 0.19; men: r(s) = -0.06, P = 0.54). CONCLUSION: There was a high level of need for information among patients with RA. Desire for involvement in treatment decision making was significantly lower and did not correlate with need for information. Associations with these needs differed for men and women. PMID- 15818717 TI - Hepatitis C virus infection in psoriatic arthritis. AB - OBJECTIVE: To evaluate the prevalence of hepatitis C virus (HCV) infection in patients with psoriatic arthritis (PsA), compared with patients affected by non HCV-related rheumatic degenerative disorders. METHODS: One-hundred consecutive subjects with PsA, and a statistically comparable group of 100 consecutive patients with peripheral osteoarthritis (OA) or sciatica due to L4-L5 or L5-S1 herniated disc were tested for HCV infection with a third-generation microparticle enzyme immunoassay (MEIA). Positive cases were submitted to a third generation recombinant immunoblot assay (RIBA) confirmatory test. Comparison between the HCV prevalence obtained in the 2 enrolled groups was performed using Fisher's exact test. RESULTS: Anti-HCV antibodies were found with the MEIA method, in 1 patient with PsA, and in 4 patients with OA or sciatica. The RIBA method confirmed MEIA results in all positive patients. The difference in HCV prevalence detected in the PsA group and in the control group was not statistically significant (P = 0.68). Furthermore, HCV prevalence in PsA patients was lower than the ones reported in different geographic areas of Italy. CONCLUSION: Our present report does not confirm previous data that indicated an increased prevalence of HCV in PsA patients, and as a consequence, does not sustain a possible trigger role of HCV in cases of PsA. The absence of clinical or instrumental resources that consent a definite differential diagnosis between PsA and HCV-related arthritis was outlined and analyzed. PMID- 15818718 TI - Indirect cost assessment in patients with rheumatoid arthritis (RA): comparison of data from the health economic patient questionnaire HEQ-RA and insurance claims data. AB - OBJECTIVE: To render information on the accuracy of patient-reported indirect cost data compared with payer-derived data of the real indirect costs on a patient-by-patient basis concerning disease-related productivity losses in rheumatoid arthritis (RA). METHODS: The assessment of indirect cost data was part of a clinical, multicenter, randomized RA trial. A total of 234 patients of working age with a diagnosis of RA (according to 1987 American College of Rheumatology criteria) were recruited. Demographics of the cohort were mean age 53 years, mean disease duration 8 years, 76% were women, and all had membership in the regional statutory health insurance plan. Every 3 months corresponding indirect cost data were derived for the cohort from a health economic questionnaire for cost assessment in patients with RA and the payer's database over a period of 18 months. Comparative statistical analyses were performed between patient-reported and insurance claims data. RESULTS: The mean annual productivity losses due to sick leave amounted to 14 and 17 days per patient (questionnaire versus payer data), and productivity losses due to work disability amounted to 3 days (both); monetary valuation renders overall costs of 1,240 and 1,590, respectively. The difference of 17% in overall productivity losses is not significant. Comparison of productivity losses reveals a strong correlation of r = 0.83 in those due to sick leave and of kappa = 0.84 in those due to work disability between questionnaire and payer data. CONCLUSION: The comparison of questionnaire and payer data shows that RA patients report their productivity losses adequately. Indirect cost assessment should therefore be included in further RA trials and observational studies. PMID- 15818719 TI - The health assessment questionnaire disability index and scleroderma health assessment questionnaire in scleroderma trials: an evaluation of their measurement properties. AB - OBJECTIVE: To evaluate the measurement properties of the Health Assessment Questionnaire (HAQ) disability index (DI) for group comparisons in scleroderma trials, and to determine if the Scleroderma Health Assessment Questionnaire (SHAQ) visual analog scales confer any measurement advantage over the HAQ DI. METHODS: A computer search for articles describing the use of the HAQ DI and SHAQ in scleroderma was performed. Evidence supporting the sensibility, reliability, validity, and responsiveness of these measures was evaluated. RESULTS: The SHAQ has incremental face and content validity over the HAQ DI because it addresses scleroderma-specific manifestations that also contribute to disability. The HAQ DI has good concurrent validity, construct validity, and predictive validity. Whether SHAQ confers incremental construct, concurrent, or predictive validity over the HAQ DI is uncertain. The HAQ DI appears more reliable than the SHAQ; however, reliability studies provide insufficient data to ascertain if minimum standards have been achieved. Responsiveness of the HAQ DI subscales has been demonstrated. CONCLUSION: The SHAQ has incremental face and content validity over the HAQ DI. The HAQ DI has greater reliability and demonstrated construct, concurrent, and predictive validity. Further investigation into the measurement properties of the HAQ DI and SHAQ visual analog scales, and their relation to the required standards of measurement is needed. PMID- 15818720 TI - Anaerobic exercise capacity in patients with juvenile-onset idiopathic inflammatory myopathies. AB - OBJECTIVE: To 1) report the feasibility of an "all-out" 30-second cycling exercise test (Wingate Anaerobic Exercise Test [WAnT]) in juvenile-onset idiopathic inflammatory myopathy (JIIM) patients, 2) describe the anaerobic exercise capacity in juvenile dermatomyositis patients, and 3) determine if the anaerobic exercise capacity could be related to disease duration or disease phase. METHODS: Twenty patients (age 14.13 +/- 5.4 years) with JIIM participated in this study. All patients were able to perform the WAnT without adverse events. RESULTS: Comparison with healthy controls revealed a -29.3 +/- 26.58% (P = 0.001) and -27.6 +/- 25.7% (P = 0.002) impairment in mean power and peak power on the WAnT, respectively. The WAnT correlated with disease phase and with knee extensor muscle strength. CONCLUSION: The WAnT might be a valuable adjunct next to other assessment tools in the followup of JIIM patients. PMID- 15818721 TI - Electron ionization induced mass spectral study of 2-alkylthio- and 4-alkykthio-5 bromouracils. PMID- 15818722 TI - Risk factors for visual loss in an Italian population-based cohort of patients with giant cell arteritis. AB - OBJECTIVE: To evaluate the frequency of visual manifestations at presentation in an Italian population-based cohort of patients with biopsy-proven giant cell arteritis (GCA), and to investigate predictors for the development of permanent visual loss. METHODS: We identified 136 Reggio Emilia (Italy) residents with biopsy-proven GCA diagnosed between 1986 and 2002. Medical records of these 136 patients were reviewed, and demographic, clinical, and laboratory data were collected.Multivariate analysis with multiple logistic regression models was performed to identify the best predictors of visual loss. RESULTS: Visual manifestations developed in 41 patients (30.1%). Partial or total visual loss was observed in 26 patients (19.1%). Anterior ischemic optic neuropathy was seen in 24 patients, and 2 patients had central retinal artery occlusion. Unilateral vision loss occurred in 19 patients, and bilateral visual loss in 7. In 25 patients, visual loss developed before glucocorticoid therapy for GCA was started. The age at disease onset was significantly higher in patients with permanent visual loss compared with those without it. The frequency of systemic signs/symptoms and erythrocyte sedimentation rate (ESR) and C-reactive protein values at diagnosis were significantly lower in patients with permanent visual loss. By multivariate logistic regression, the only statistically significant predictor for the development of permanent visual loss was the absence of high levels of ESR at diagnosis (tertile 2: Odds ratio [OR] 0.08; tertile 3: OR 0.11). Other predictors included in the model were the absence of systemic manifestations (OR 0.24), an older age at disease diagnosis (quintile 5: OR 5.60), and the presence of an elevated platelet count at diagnosis (OR 4.99), however they were only of borderline statistical significance. CONCLUSION: The proportion of Italian patients with GCA that developed visual loss was similar to that reported from other countries. The patients with low inflammatory response had a higher risk of visual loss. PMID- 15818723 TI - Study of peptide on-line complexation with transition-metal ions generated from sacrificial electrodes in thin-chip polymer microsprays. AB - A miniaturized polymer electrospray-type interface is used to study metal-ion chelation with model peptides. Taking advantage of the intrinsic electrochemical behavior of electrospray, a sacrificial electrode is used to generate at the same time electrospray and transition-metal ions coming from the anodic dissolution of the electrode. The microspray interface provides enhanced mass transport due to its small dimensions, increasing the yield of possible reactions, in particular complex formation. Transition-metal electrodes, e.g. copper, zinc, nickel, iron and silver, are used to obtain on-line complexation with model peptides. It is demonstrated that the use of in-reservoir sacrificial electrodes is an efficient way to generate metal ions in order to form and study complexes with peptides, avoiding the addition of metallic salts. PMID- 15818724 TI - Validation of a high-throughput absorption, distribution, metabolism, and excretion (ADME) system and results for 60 literature compounds. AB - A high-throughput analytical system for the support of absorption, distribution, metabolism, and excretion (ADME) was constructed in collaboration with Gubbs Inc. to commercialize corresponding software. We sought to quickly build and validate this system for microsomal clearance assessment using 60 commercially available non-proprietary compounds that are non-DEA-restricted in addition to 36 proprietary Millennium compounds that had already been assessed using a low throughput infrastructure. The system was constructed such that a approximately 45 second total cycle time was achieved injection-to-injection. Software was successfully coded to enable the analyst to submit multiple batches, and modify multiple methods very quickly for use with Applied Biosystems Analyst 1.3. After acquisition the software was used to simultaneously integrate multiple injection chromatograms, regress the data, and calculate clearance such that all of the data could be easily and immediately reviewed by both bioanalytical and enzymology personnel. Unfortunately, despite an exhaustive search of the literature, we were unable to find a large number of non-proprietary compound data for validation, so we provide such a source of data here. Results are presented for the 60 literature compounds that were assessed. A good correlation was observed between literature results for 16 compounds that we were able to find and the results obtained using the system. The Millennium proprietary compounds that we assessed using both low- and high-throughput approaches also correlated well. We present here a system for the support of high-throughput in vitro ADME analysis and also present the results of 60 non-proprietary, non-DEA scheduled compounds to facilitate the validation efforts of others. Finally, we present commercially available software to facilitate high-throughput ADME systems in the community. PMID- 15818725 TI - Bayesian model choice and infection route modelling in an outbreak of Norovirus. AB - A data set from an outbreak of gastroenteritis in a school is analysed using a stochastic transmission model. The causative agent of the outbreak is believed to be a Norovirus, spread through person-to-person contact. Particular attention is given to the question of whether or not vomiting episodes enhance the spread of the virus via aerosol transmission. The methodology developed uses Bayesian model choice, implemented with reversible-jump Markov chain Monte Carlo methods. The methodology appears to be highly sensitive to assumptions made concerning the data, which provides some assurance that the conclusions are driven by observations rather than the underlying model and methodology. PMID- 15818726 TI - Determination of 2-methoxyestradiol in human plasma, using liquid chromatography/tandem mass spectrometry. AB - A liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay was developed for the quantitative determination of 2-methoxyestradiol (2ME2) in human plasma. Sample pretreatment involved liquid-liquid extraction with ethyl acetate of 0.3 mL aliquots of plasma spiked with the internal standard, deuterated 2ME2 (2ME2 d5). Separation was achieved on a Zorbax Eclipse C18 column (2.1 x 50 mm, i.d., 5 microm) at room temperature using a gradient elution with methanol and water at a flow rate of 0.25 mL/min. Detection was performed using atmospheric pressure chemical ionization MS/MS by monitoring the ion transitions from m/z 303.1 --> 136.8 (2ME2) and m/z 308.1 --> 138.8 (2ME2-d5). Calibration curves were linear in the concentration range of 1-100 ng/mL. The accuracy and precision values, obtained from three different sets of quality control samples analyzed in quintuplicate on four separate occasions, ranged from 105-108% and from 3.62 5.68%, respectively. This assay was subsequently used for the determination of 2ME2 concentration in plasma of a patient with cancer after a single oral administration of 2ME2 at a dose of 2200 mg. PMID- 15818727 TI - Surgical resection for esophageal carcinoma: speaking the language. AB - The terminology used to describe esophagectomy for carcinoma can be confusing, even for specialists in gastrointestinal disease. As a result, specific terms are often used out of their intended context. To simplify the nomenclature, two points regarding procedures for surgical resection of the esophagus are critical: the extent of resection (radical vs standard) and the operative approach (choice of incisions). It is important to understand that the radicality of the resection may have little to do with the operative approach, with the exception of esophagectomy without thoracotomy (transhiatal esophagectomy), which mandates the performance of a standard or non-radical resection. Esophagectomy has emerged as the standard curative treatment option for patients with esophageal carcinoma; however, unlike the surgical resection of other types of solid tumors, many different surgical options and/or approaches exist for these patients. This heterogeneity of care may result from the fact that the esophagus is accessible through more than one body cavity (left hemithorax, right hemithorax, abdomen). In addition, and partially as a result of its accessibility, different types of surgical specialists harbor this operation in their armamentarium, including general surgeons, thoracic surgeons, and surgical oncologists. Despite this enthusiasm amongst surgeons, little consensus exists as to which option is most oncologically sound. Further, the details of the various surgical approaches and procedures for resection of the esophagus are often difficult to comprehend, even for specialists in gastrointestinal disease, with much of the relevant terminology used out of its intended context. To facilitate the understanding of the surgical options for esophageal carcinoma, it is useful to view the operation from two angles: the extent of resection (Aradical@ vs Astandard@) and the operative approach (choice of incisions). PMID- 15818728 TI - Transcription factor Sp1 expression in gastric cancer and its relationship to long-term prognosis. AB - AIM: To explore the expression of Sp1 in gastric carcinoma as well as its association with other clinicopathologic features, and to evaluate the role of Sp1 as a prognostic indicator of gastric carcinoma. METHODS: By using immunohistochemistry, we examined the Sp1 expression patterns in 65 cases of human gastric cancer, and 40 normal gastric mucosa specimens. Simultaneously, the correlation between Sp1 expression and clinical outcome or clinicopathologic features was investigated. RESULTS: The percentage of Sp1 expression was 12.5% (5/40) in normal gastric mucosa, and the Sp1 protein was mainly expressed in the nuclei of cells located in the mucous neck region. In sharp contrast, strong Sp1 expression was detected in tumor cells, whereas no or faint Sp1 staining was detected in stromal cells and normal glandular cells surrounding the tumors. The expression rate of Sp1 in gastric cancer lesions was 53.85% (35/65). The medium survival duration in patients who had a tumor with negative, weak and strong Sp1 expressions was 1 700, 1 560 and 1 026 d, respectively (P<0.05). Sp1 protein expression was closely related to the depth of tumor infiltration (chi(2) = 13.223, P<0.01) and TNM stage (chi(2) = 11.009, P<0.05), but had no relationship with the number of lymph nodes and Lauren's classification (P>0.05). Cox regression model for multivariate analysis revealed that high Sp1 expression (P<0.05) and advanced stage (P<0.01) were independent predictors of poor survival. CONCLUSION: Normal and malignant gastric tissues have unique Sp1 expression patterns. Sp1 might serve as an independent prognostic factor, by influencing the tumor infiltration and progression. PMID- 15818729 TI - Expression of p16 gene and Rb protein in gastric carcinoma and their clinicopathological significance. AB - AIM: To analyze the correlation between the protein expression of p16 and Rb genes in gastric carcinoma (GC), to investigate the role of p16 gene in invasion and lymph node metastasis of GC, and to examine the deletion and mutation in exon 2 of p16 gene in GC. METHODS: The protein expression of p16 and Rb genes was examined by streptavidin-peroxidase conjugated method (S-P) in normal gastric mucosa, dysplastic gastric mucosa and GC. The deletion and mutation of p16 gene were examined by polymerase chain reaction (PCR) and polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) respectively in normal gastric mucosa and GC. RESULTS: The positive rates of P16 and Rb protein expression respectively were 96% (77/80) and 99% (79/80) in normal gastric mucosa, 92% (45/50) and 80% (40/50) in dysplastic gastric mucosa, 48% (58/122) and 60% (73/122) in GC. The positive rates of P16 and Rb protein expression in GC were significantly lower than that in normal gastric mucosa and dysplastic gastric mucosa (P<0.05). The positive rate of P16 protein expression in mucoid carcinoma (10%, 1/10) was significantly lower than that in poorly differentiated carcinoma (51%, 21/41), undifferentiated carcinoma (58%, 15/26) and signet ring cell carcinoma (62%, 10/16) (P<0.05). The positive rates of P16 protein in 30 cases of paired primary and lymph node metastatic GC were 47% (14/30) and 17% (5/30) respectively, being significantly lower in the later than in the former (P<0.05). There was no mutation in exon 2 of p16 gene in the 25 freshly resected primary GCs. But five cases in the 25 freshly resected primary GCs displayed deletion in exon 2 of p16 gene. The positive rate of both P16 and Rb proteins was 16% (14/90), and the negative rate of both P16 and Rb proteins was 8% (7/90) in 90 GCs. The rate of positive P16 protein with negative Rb protein was 33% (30/90). The rate of negative P16 protein with positive Rb protein was 43% (39/90). There was reverse correlation between P16 and Rb expression in 90 GCs (P<0.05). CONCLUSION: The loss protein expression of p16 and Rb genes is related to GC. The loss expression of P16 protein is related to the histopathologic subtypes and lymph node metastasis of GC. Expression of P16 and Rb proteins in GC is reversely correlated. The deletion but not mutation in exon 2 of p16 gene may be involved in GC. PMID- 15818730 TI - Suppression of gastric cancer growth by adenovirus-mediated transfer of the PTEN gene. AB - AIM: To investigate the tumor-suppressive effect of the phosphatase and tensin homologue deleted from chromosome (PTEN) in human gastric cancer cells that were wild type for PTEN. METHODS: Adenoviruses expressing PTEN or luciferase as a control were introduced into gastric cancer cells. The effect of exogenous PTEN gene on the growth and apoptosis of gastric cancer cells that are wtPTEN were examined in vitro and in vivo. RESULTS: Adenovirus-mediated transfer of PTEN (Ad PTEN) suppressed cell growth and induced apoptosis significantly in gastric cancer cells (MGC-803, SGC-7901) carrying wtPTEN in comparison with that in normal gastric epithelial cells (GES-1) carrying wtPTEN. This suppression was induced through downregulation of the Akt/PKB pathway, dephosphorylation of focal adhesion kinase and mitogen-activated protein kinase and cell-cycle arrest at the G2/M phase but not at the G1 phase. Furthermore, treatment of human gastric tumor xenografts (MGC-803, SGC-7901) with Ad-PTEN resulted in a significant (P<0.01) suppression of tumor growth. CONCLUSION: These results indicate a significant tumor-suppressive effect of Ad-PTEN against human gastric cancer cells. Thus, Ad PTEN may be used as a potential therapeutic strategy for treatment of gastric cancers. PMID- 15818731 TI - Inhibitory effect of human telomerase antisense oligodeoxyribonucleotides on the growth of gastric cancer cell lines in variant tumor pathological subtype. AB - AIM: To investigate the inhibitory effect of specialized human telomerase antisense oligodeoxyribonucleotides on the growth of well (MKN-28), moderately (SGC-7901) and poorly (MKN-45) differentiated gastric cancer cell lines under specific conditions and its inhibition mechanism, and to observe the correlation between the growth inhibition ratio and the tumor pathologic subtype of gastric cancer cells. METHODS: Telomerase activity in three gastric cancer cell lines of variant tumor pathologic subtype was determined by modified TRAP assay before and after the specialized human telomerase antisense oligodeoxyribonucleotides were dealt with under specific conditions. Effect of antisense oligomer under specific conditions of the growth and viability of gastric cancer cell lines was explored by using trypan blue dye exclusion assay, and cell apoptosis was detected by cell morphology observation, flow cytometry and TUNEL assay. RESULTS: Telomerase activity was detected in well, moderately and poorly differentiated gastric cancer cell lines (the quantification expression of telomerase activity was 43.7TPG, 56.5TPG, 76.7TPG, respectively). Telomerase activity was controlled to 30.2TPG, 36.3TPG and 35.2TPG for MKN-28, SGC-7901 and MKN-45 cell lines respectively after treatment with human telomerase antisense oligomers at the concentration of 5 mumol/L, and was entirely inhibited at 10 mumol/L, against the template region of telomerase RNA component, whereas no inhibition effect was detected in missense oligomers (P<0.05). After treatment with antisense oligomers at different concentrations under specific conditions for 96 h, significant growth inhibition effects were found in MKN-45 and SGC-7901 gastric cancer cell lines (the inhibition ratio was 40.89% and 71.28%), but not in MKN-28 cell lines (15.86%). The ratio of inactive SGC-7901 cells increased according to the prolongation of treatment from 48 to 96 h. Missense oligomers could not lead to the same effect (P<0.05). Apoptosis of SGC-7901 and MKN-45 cells was detected not only by morphology and TUNEL assay but also by flow cytometry. The apoptotic rate reached 33.56% for SGC-7901 cells and 44.75% for MKN-45 cells. CONCLUSION: The viability and proliferation of gastric cancer cells can be inhibited by antisense telomerase oligomers. The growth inhibition of gastric cancer cells is correlated with concentration, time and sequence specialty of antisense oligomers. The inhibition mechanism of antisense human telomerase oligomers depends not only on the sequence specialty but also on the biological characteristics of gastric cancer cell lines. PMID- 15818732 TI - Hepatitis B virus genotypes and hepatocellular carcinoma in Thailand. AB - AIM: The role of hepatitis B virus (HBV) genotypes on the clinical features and prognosis of patients with hepatocellular carcinoma (HCC) is currently unknown. The aim of the present study was to evaluate the distribution of HBV genotypes and their clinical relevance in Thai patients. METHODS: HBV genotypes were determined by PCR-RFLP in stored sera of 93 asymptomatic carriers, 103 patients with chronic hepatitis, 60 patients with cirrhosis and 76 patients with HCC. The clinical data were analyzed in relation to the HBV genotype. RESULTS: HBV genotypes C and B were predominant in Thailand, accounting for 73% and 21%, respectively. The distributions of genotypes B and C were similar in HCC patients compared to the other groups. Genotype C was significantly more common in HCC patients who were under 40 years old than genotype B (18% vs 0%, P = 0.03), but was significantly less common in patients older than 60 years (26% vs 56.5%, P = 0.01). The positive rate of hepatitis B e antigen (HBeAg) in patients with genotype C was significantly higher than that in patients with genotype B (71.6% vs 44.4%, P = 0.03 in chronic hepatitis; 56.8% vs 11.1%, P = 0.01 in cirrhosis). There were no differences between HCC patients with genotypes B and C regarding tumor staging by CLIP criteria and the overall median survival. Multivariate analyses showed that HBV genotype was not an independent prognostic factor of survival in HCC patients. CONCLUSION: Patients with genotype C had a higher positive rate of HBeAg and exhibited earlier progression of cirrhosis and HCC than those with genotype B. However, there were no differences in the risk of developing HCC and its prognosis between patients with these genotypes. PMID- 15818733 TI - Ornithine decarboxylase gene is overexpressed in colorectal carcinoma. AB - AIM: To investigate the ornithine decarboxylase (ODC) gene expression in colorectal carcinoma, ODC mRNA was assayed by RT-PCR and ODC protein was detected by a monoclonal antibody against fusion of human colon ODC prepared by hybridoma technology. METHODS: Total RNA was extracted from human colorectal cancer tissues and their normal counterpart tissues. ODC mRNA levels were examined by RT-PCR. ODC genes amplified from RT-PCR were cloned into a prokaryotic vector pQE-30. The expressed proteins were purified by chromatography. Anti-ODC mAb was prepared with classical hybridoma techniques and used to determine the ODC expression in colon cancer tissues by immunohistochemical and Western blotting assay. RESULTS: A cell line, which could steadily secrete anti-ODC mAb, was selected through subcloning four times. Western blotting reconfirmed the mAb and ELISA showed that its subtype was IgG2a. RT-PCR showed that the ODC mRNA level increased greatly in colon cancer tissues (P<0.01). Immunohistochemical staining showed that colorectal carcinoma cells expressed a significantly higher level of ODC than normal colorectal mucosa (98.6+/-1.03% vs 5.26+/-5%, P<0.01). CONCLUSION: ODC gene overexpression is significantly related to human colorectal carcinoma. ODC gene expression may be a marker for the gene diagnosis and therapy of colorectal carcinoma. PMID- 15818734 TI - Rectosigmoid findings are not associated with proximal colon cancer: analysis of 6 196 consecutive cases undergoing total colonoscopy. AB - AIM: To review the risk of proximal colon cancer in patients undergoing colonoscopy. METHODS: We estimated the risk of advanced proximal adenomas and cancers in 6 196 consecutive patients that underwent colonoscopy (mean age 60 years, 65% males, without prior history of colorectal examination). Neoplasms were classified as diminutive adenoma (5 mm or less), small adenoma (6-9 mm), advanced adenoma (10 mm or more, with villous component or high-grade dysplasia) and cancer (invasive adenocarcinoma). The sites of neoplasms were defined as rectosigmoid (rectum and sigmoid colon) and proximal colon (from cecum to descending colon). RESULTS: The trend of the prevalence of advanced proximal adenoma was to increase with severe rectosigmoid findings, while the prevalence of proximal colon cancer did not increase with severe rectosigmoid findings. Among the 157 patients with proximal colon cancer, 74% had no neoplasm in the rectosigmoid colon. Multivariate logistic-regression analysis revealed that age was the main predictor of proximal colon cancer and existence of rectosigmoid adenoma was not a predictor of proximal colon cancer. CONCLUSION: Sigmoidoscopy is inadequate for colorectal cancer screening, especially in older populations. PMID- 15818735 TI - In vitro growth inhibition of human colonic tumor cells by Verapamil. AB - AIM: To investigate the effects and mechanisms of Verapamil on cultured human colonic tumor (HCT) cells. METHODS: HCT cells were treated with different concentrations of Verapamil, and their proliferation was examined by MTT assay. The areas of sub-diploid peak were measured by flow cytometry, and the DNA ladder was found by agarose gel electrophoresis. The characteristic changes in morphology were observed under light microscopy. The cell nuclei (propidium iodide labeled, PI-labeled) and cellular distribution and concentration of calcium (Fluo-3-labeled) were studied by using laser confocal scanning microscope. RESULTS: The proliferation of HCT cells was inhibited by different concentrations of Verapamil. With the increase in concentration of Verapamil, the percent of G0-G1 phase cells in HCT cells increased and that of S phase cells decreased. After treating with different concentrations of Verapamil, flow cytometry showed that HCT cells were enlarged in areas of sub-diploid in a dose dependent manner. Gel electrophoresis results displayed a typical DNA ladder. On staining with Wrights-Giemsa, the typical cellular apoptosis morphologic changes were also observed. PI-labeled cell nuclei were found markedly changed. In addition, we inspected that the 100 micromol/L Verapamil could increase the intracellular calcium ion concentration [Ca(2+)](i) in HCT cells. CONCLUSION: Verapamil can inhibit proliferation of HCT cells via inducing cell apoptosis. PMID- 15818736 TI - Expression of Helicobacter pylori AlpA protein and its immunogenicity. AB - AIM: To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori (H pylori) and to study the immunogenicity of adhesin AlpA. METHODS: Gene Ab, which was amplified from H pylori chromosomal DNA by PCR technique, was sequenced and the biological information was analyzed, and inserted into the Nco I and Not I restriction fragments of the expression vector pET-22b(+) using T4 DNA ligase. The resulting plasmid pET-AlpA was transformed into competent E.coli BL21(DE3) cells using ampicillin resistance for selection. Recombinant strains were incubated in 5 mL LB with 100 mug/mL ampicillin overnight at 37 degrees. Sonication of BL21(DE3)pET-22b(+)/AlpA was analyzed by Western blot to detect AlpA immunogenicity. RESULTS: The gene encoding AlpA protein was amplified by PCR with chromosomal DNA of H pylori Sydney strain (SS1) as templates. It revealed that AlpA DNA fragment amplified by PCR had approximately 1 500 nucleotides, compatible with the previous reports. The recombinant plasmid pET-22b(+)/AB was successfully constructed. DNA sequencing showed one open reading frame with the length of 588 bp. It encoded seven conservative regions that showed good antigenicity and hydrophobicity by Parker and Welling method. Furthermore, INTERNET EXPASY, NNPREDICT and ISREC predicted that it was a porin-like structure consisting of beta-pleated sheets that were embedded in the outer membrane. BLAST analyzed 836 767 protein sequences and found that the similar sequences were all belonging to H pylori OMP sequences. SDS-PAGE and scan analysis showed that the molecular weight of AB was 22.5 ku and recombinant protein amounted to 29% of the total bacterial protein, among which dissolved expression amounted to 21.9% of sonicated supernatant. The rAB purity amounted to 96% through affinity chromatography. Western blot analysis of rAB confirmed that it could be specially recognized by serum form rabbit immunized with AlpA and H pylori infected. CONCLUSION: Adhesin AlpA recombinant protein may be a potential vaccine for control and treatment of H pylori infection. PMID- 15818737 TI - Effects of fibril- or fixed-collagen on matrix metalloproteinase-1 and tissue inhibitor of matrix metalloproteinase-1 production in the human hepatocyte cell line HLE. AB - AIM: Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic stellate cells (HSC), but not in hepatocytes. We investigated the effects of fibril- and fixed-collagen on MMP-1 and TIMP-1 production in hepatocytes, using the HLE cell line. METHODS: Fibril type I and IV collagen were prepared by HCl digestion of type I and IV collagen, respectively. For fixed-collagen, culture dishes were coated with fibril type I or IV collagen and fixed by ultraviolet. Type I collagenase activity was measured using fluorescein isothiocyanate-labeled type I collagen. MMP-1 and TIMP-1 in HLE cells were measured by a one-step sandwich enzyme immunoassay. RESULTS: Both fibril type I and IV collagen significantly increased type I collagenase activity about two-fold compared with no fibril collagen. The effects of the fibril collagen were not affected by the coating condition. There was no significant difference in the effects on collagenase activity between cells cultured in medium containing fibril type I collagen and those cultured in the presence of type IV collagen. Both types of fibril collagen significantly increased MMP-1 production, and showed more than 10-fold higher levels of MMP-1 than the control. The enhanced MMP-1 production by fibril collagens was unaffected by the coating condition. By contrast, TIMP-1 production was not changed by the addition of fibril type I or IV collagen, and neither was it affected by the coating conditions. Coating with type I collagen significantly suppressed MMP-1 production by almost one-tenth compared with no coating. By contrast, TIMP-1 production was not affected by either the absence of a collagen coat or by increasing the concentration of the coating collagen. CONCLUSION: These results indicated that, in HLE cells, fibril- and fixed-collagen have opposite effects on MMP-1 production without affecting TIMP production. Fibril collagen induced collagenase activity by up-regulation of MMP-1 production without affecting TIMP-1 production. By contrast, fixed collagen reduced MMP-1 production. Our results suggest that hepatocytes might also play an important role in the regulation of the hepatic fibrosis alongside HSC. PMID- 15818738 TI - Effects of Chinese traditional compound, JinSanE, on expression of TGF-beta1 and TGF-beta1 type II receptor mRNA, Smad3 and Smad7 on experimental hepatic fibrosis in vivo. AB - AIM: The transforming growth factor-beta (TGF-beta)/Smad signaling pathway system plays a prominent role in the control of cell growth and extracellular matrix formation in the progression of liver fibrogenesis. Smad proteins can either positively or negatively regulate TGF-beta responses. In this study, the therapeutic effects of Chinese traditional compound decoction, JinSanE, and the changes of TGF-beta/Smad signaling pathway system in carbon tetrachloride (CCl(4))-induced rat experimental liver fibrosis were examined. METHODS: Seventy two healthy Wistar rats were assigned to groups including normal control group, CCl(4) model group, JinSanE treatment group I and JinSanE treatment group II. Each group contained 18 rats. All groups, except the normal control group, received CCl(4) subcutaneous injection for 8 wk. Rats in JinSanE groups I and II were orally treated with JinSanE daily at the 1(st) and 5(th) wk, respectively, after exposure to CCl(4). The expression of TGF-beta1 and TGF-beta1 type II receptor (TRII) mRNA in the liver was determined by reverse transcription polymerase chain reaction, and the expression of TGF-beta1, Smad3 and Smad7 by immunohistochemistry. The liver histopathology was also examined by HE staining and observed under electron microscope. The activities of several serum fibrosis associated enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), the levels of serum hyaluronic acid (HA) were assayed. RESULTS: Hepatic fibrosis caused by CCl(4) was significantly inhibited in the JinSanE-treated groups. The degrees of necrosis/degeneration and fibrosis scores were significantly lower in the JinSanE-treated groups than in the model control group. The expression of TGF-beta1, TRII and Smad3 was significantly higher in the model group than that in the JinSanE-treated groups, and the active/total TGF beta1 ratio in the JinSanE groups was suppressed. Expression of TRII mRNA and Smad3 proteins showed a distribution pattern similar to that of TGF-beta1 with a direct correlation in terms of the degree of hepatic fibrosis. The amount of positive staining Smad7 cells was significantly less in the model group than in the JinSanE-treated groups and the normal group. The contents of ALT, AST and HA were significantly lower in the JinSanE-treated groups than those in the model group. CONCLUSION: Traditional Chinese medicine, JinSanE, prevents the progression of hepatic damage and fibrosis through the inhibition of TGF-beta1, TRII and Smad3 signal proteins, and increases expression of Smad7 signal protein in vivo. PMID- 15818739 TI - Heterogeneity in predisposition of hepatic cells to be induced into pancreatic endocrine cells by PDX-1. AB - AIM: The role of Pancreatic and Duodenal Homeobox-1 (PDX-1) as a major regulator of pancreatic development determines the function and phenotype of beta cell. In this study, potential plasticity of liver cells into pancreatic endocrine cells induced by PDX-1 was evaluated. METHODS: Human hepatoma cell line HepG2 was stably transfected with mammalian expression plasmid pcDNA3-PDX encoding human PDX-1 gene. Ectopic expression of PDX-1 and insulin were detected by RT-PCR, Western blot and/or immunostaining. PDX-1(+) HepG2 cells were transplanted under renal capsule of STZ-induced diabetic nude mice (n = 16) to examine the inducing effect in vivo. RESULTS: Exogenous PDX-1 transgene was proved to express effectively in HepG2 cell at both mRNA and protein levels. The expression of endogenous insulin and some beta cell-specific differentiation markers and transcription factors were not induced in PDX-1(+) HepG2 cells. When transplanted under renal capsule of STZ-induced diabetic nude mice, PDX-1(+) HepG2 cells did not generate insulin-producing cells. These data indicated that stable transfected PDX-1 could not convert hepatoma cell line HepG2 to pancreatic cells in vitro or in vivo. Mature hepatocytes might need much more complicated or rigorous conditions to be shifted to insulin-producing cells. CONCLUSION: The expression of exogenous PDX-1 is not sufficient to induce relatively mature hepatocytes differentiating into insulin-producing cells. PMID- 15818740 TI - Differential gene expression during capillary morphogenesis in a microcarrier based three-dimensional in vitro model of angiogenesis with focus on chemokines and chemokine receptors. AB - AIM: To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogenesis system with affymetrix oligonucleotide array. METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed. RESULTS: A total of 1 961 genes were up- or down-regulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-alpha, CXCL2/GRO-beta, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, CX3CL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis. CONCLUSION: The present study demonstrates a global profile of gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be conducted. PMID- 15818741 TI - Expression of intestinal trefoil factor, proliferating cell nuclear antigen and histological changes in intestine of rats after intrauterine asphyxia. AB - AIM: To study the expressions of intestinal trefoil factor (ITF) and proliferating cell nuclear antigen (PCNA) and histologic changes in intestine, to investigate the relationship between ITF and intestinal damage and repair after intrauterine hypoxia so as to understand the mechanism of intestinal injury and to find a new way to prevent and treat gastrointestinal diseases. METHODS: Wistar rats, pregnant for 21 d, were used to establish animal models of intrauterine asphyxia by clamping one side of vessels supplying blood to uterus for 20 min, another side was regarded as sham operation group. Intestinal tissues were taken away at 0, 24, 48 and 72 h after birth and stored in different styles. ITF mRNA was detected by RT-PCR. PCNA expression was measured by immunohistochemistry. Intestinal tissues were studied histologically by HE staining in order to observe the areas and degree of injury and to value the intestinal mucosa injury index (IMDI). RESULTS: ITF mRNA appeared in full-term rats and increased with age. After ischemia, ITF mRNA was decreased to the minimum (0.59+/-0.032) 24 h after birth, then began to increase higher after 72 h than it was in the control group (P<0.01). PCNA positive staining located in goblet cell nuclei. The PCNA level had a remarkable decline (53.29+/-1.97) 48 h after ischemia. Structure changes were obvious in 48-h group, IMDI (3.40+/-0.16) was significantly increased. Correlation analyses showed that IMDI had a negative correlation with ITF mRNA and PCNA (r = -0.543, P<0.05; r = -0.794, P<0.01, respectively). CONCLUSION: Intrauterine ischemia can result in an early decrease of ITF mRNA expression. ITF and PCNA may play an important role in the damage and repair of intestinal mucosa. PMID- 15818742 TI - Identification of expressed genes in regenerating rat liver in 0-4-8-12 h short interval successive partial hepatectomy. AB - AIM: To identify the genes differentially expressed in the regenerating rat liver of 0-4-8-12 h short interval successive partial hepatectomy (SISPH) and to analyze their expression profiles. METHODS: Five hundred and fifty-one elements screened from subtractive cDNA libraries were made into a cDNA microarray (cDNA chip). Extensive gene expression analysis following 0-4-8-12 h SISPH was conducted by microarray. RESULTS: One hundred and eighty-three elements were selected, which were either up- or down-regulated more than 2-fold at one or more time points after SISPH. Cluster analysis and generalization analysis showed that there were five distinct temporal patterns of gene expression. Eighty-six genes were unreported, associated with liver regeneration (LR). CONCLUSION: Microarray analysis shows that the down regulated genes are much more than the up-regulated ones in SISPH; the numbers of genes expressed consistently are fewer than that expressed immediately; the genes expressed in high abundance are much fewer than that increased 2-5-fold. The comparison of SISPH with partial hepatectomy (PH) shows that the expression trends of most genes in SISPH and in PH are similar, but the expression of 43 genes is specifically altered in SISPH. PMID- 15818743 TI - Vascular contractile response and signal transduction in endothelium-denuded aorta from cirrhotic rats. AB - AIM: The mechanism of decreased vascular reactivity to vasoconstrictors in portal hypertension is still unclear. In addition to nitric oxide, defects in post receptor signal transduction pathway have been suggested to play a role. However, substantial evidences observed equivocal changes of vascular reactivity following different agonists that challenged the hypothesis of the post-receptor defect. The current study was to evaluate the vascular reactivity to different agonists and the inositol trisphosphate (IP(3)) changes in signal transduction cascade from cirrhotic rats with portal hypertension. METHODS: The endothelial denuded aortic rings from cirrhotic and sham-operated rats were obtained for ex vivo tension study and measurement of the corresponding [(3)H] IP(3) formation following different receptor and nonreceptor-mediated agonists' stimulation. Additionally, iNOS protein expression was measured in thoracic aorta. The contractile response curves to phenylephrine were performed in endothelial denuded aortic rings with and without preincubation with a specific iNOS inhibitor (L-N(6)-(1-iminoethyl)-lysine, L-NIL). RESULTS: In endothelial denuded aortic rings of cirrhotic rats, the vascular responses were reduced with phenylephrine and arginine vasopressin (AVP) stimulation but were normal with U 46619, NaF/AlCl(3), and phorbol esterdibutyrate (PdBU) stimulation. Compared to the corresponding control groups, the degree of the increment of [(3)H] IP(3) formation from basal level was also decreased with phenylephrine and AVP stimulation, but was normal with U-46619 and NaF/AlCl(3) stimulation. The preincubation with L-NIL did not modify the hyporesponsiveness to phenylephrine. Additionally, the iNOS protein expression in thoracic aorta was not different in cirrhotic and sham-operated rats. CONCLUSION: Without the influence of nitric oxide, vascular hyporeactivity to vasoconstrictors persisted in cirrhotic rats with portal hypertension. However, the decreased vascular reactivity is an agonist-specific phenomenon. In addition, G-protein and phospholipase C pathway associated with the IP(3) productions may be intact in cirrhotic rats with portal hypertension. PMID- 15818744 TI - Impact of comorbid anxiety and depression on quality of life and cellular immunity changes in patients with digestive tract cancers. AB - AIM: A study was performed to investigate the impact of comorbid anxiety and depression (CAD) on quality of life (QOL) and cellular immunity changes in patients with digestive tract cancers. METHODS: One hundred and fifty-six cases of both sexes with cancers of the digestive tract admitted between March 2001 and February 2004 in the Department of Medical Oncology, First Affiliated Hospital of Xi'an Jiaotong University were randomly enrolled in the study. Depressive and anxiety disorder diagnoses were assessed by using the Structured Clinical Interview for DSM-IV. All adult patients were evaluated with the Hamilton depressive scale (HAMD, the 24-item version), the Hamilton anxiety scale (HAMA, a modified 14-item version), quality of life questionnaire-core 30 (QLQ-C30), social support rating scale (SSRS), simple coping style questionnaire (SCSQ), and other questionnaires, respectively. In terms of HAMD > or = 20 and HAMA > or = 14, the patients were categorized, including CAD (n = 31) in group A, anxiety disorder (n = 23) in group B, depressive disorder (n = 37) in group C, and non disorder (n = 65) in group D. Immunological parameters such as T-lymphocyte subsets and natural killer (NK) cell activities in peripheral blood were determined and compared among the four groups. RESULTS: The incidence of CAD was 21.15% in patients with digestive tract cancers. The average scores of social support was 43.67+/-7.05 for 156 cases, active coping 20.34+/-7.33, and passive coping 9.55+/-5.51. Compared with group D, subjective support was enhanced slightly in group A, but social support, objective support, and utilization of support reduced, especially utilization of support with significance (6.16 vs 7.80, P<0.05); total scores of active coping decreased, while passive coping reversed; granulocytes proliferated, monocytes declined, and lymphocytes declined significantly (32.87 vs 34.00, P<0.05); moreover, the percentage of CD3, CD4, CD8 and CD56 in T lymphocyte subsets was in lower level, respectively, and CD56 showed a significant decline in group A (26.02 vs 32.20, P<0.05), however, CD4/CD8 ratio increased. Physical function, role function, fatigue, sleeplessness and constipation had significant changes among different groups by one-way ANOVA, and group A was in poor QOL. It revealed that global health-related quality of life (QL) were positively correlated with active coping and CD56; CAD was negatively correlated with QL, active coping and CD56. Furthermore, the step-wise regression analysis suggested that utilization of support, CD56, active coping, fatigue, sleeplessness and depression were significant factors contributing to QOL. CONCLUSION: CAD, which can impair QOL and cellular immunity, occurs with a higher incidence in patients with digestive tract cancers. Hence, it is essential to improve mental health for them with specifically tailored interventions. PMID- 15818745 TI - Nitroester drug's effects and their antagonistic effects against morphine on human sphincter of Oddi motility. AB - AIM: To evaluate the effects of nitroester drugs on human sphincter of Oddi (SO) motility and their antagonistic effects against morphine which shows excitatory effect on Oddi's sphincter motility. METHODS: The effects of these drugs on SO were evaluated by means of choledochofiberoscopy manometry. A total of 67 patients having T-tubes after cholecystectomy and choledochotomy were involved in the study, they were randomly divided into glyceryl trinitrate (GTN) group, isosorbide dinitrate (ISDN) group, pentaerythritol tetranitrate (PTN) group, morphine associated with GTN group, morphine associated with ISDN group and morphine associated with PTN group. Basal pressure of Oddi's sphincter (BPOS), amplitude of phasic contractions (SOCA), frequency of phasic contractions (SOF), duration of phasic contractions (SOD), duodenal pressure (DP) and common bile duct pressure (CBDP) were scored and analyzed. Morphine was given intramuscularly while nitroester drugs were applied sublingually. RESULTS: BPOS and SOCA decreased significantly after administration of ISDN and GTN, BPOS reduced from 10.95+/-7.49 mmHg to 5.92+/-4.04 mmHg (P<0.05) evidently after application of PTN. BPOS increased from 7.37+/-5.58 mmHg to 16.60+/-13.87 mmHg, SOCA increased from 54.09+/-38.37 mmHg to 100.70+/-43.51 mmHg, SOF increased from 7.15+/-3.20 mmHg to 10.38+/-2.93 mmHg and CBDP increased 3.75+/-1.95 mmHg to 10.49+/-8.21 mmHg (P<0.01) evidently after injection of morphine. After associated application of ISDN and GTN, the four indications above decreased obviously. As for application associated with PTN, SOCA and SOF decreased separately from 100.64+/ 44.99 mmHg to 66.17+/-35.88 mmHg and from 10.70+/-2.76 mmHg to 9.04+/-1.71 mmHg (P<0.05) markedly. CONCLUSION: The regular dose of GTN, ISDN and PTN showed inhibitory effect on SO motility, morphine showed excitatory effect on SO while GTN, ISDN and PTN could antagonize the effect of morphine. Among the three nitroester drugs, the effect of ISDN on SO was most significant. PMID- 15818746 TI - Multi-detector CT enterography with iso-osmotic mannitol as oral contrast for detecting small bowel disease. AB - AIM: To assess the feasibility and usefulness of multi-detector CT enterography with orally administered iso-osmotic mannitol as negative contrast in demonstrating small bowel disease. METHODS: Thirteen volunteers and 38 patients with various kinds of small bowel disease were examined. We administered about 1 500 mL iso-osmotic mannitol as negative contrast agent and then proceeded with helical CT scanning on a Siemens Sensation 16 scanner. All volunteers and patients were interviewed about their tolerance of the procedure. Two radiologists post-processed imaging data with MPR, thin MIP, VRT and INSPACE when necessary and then interpreted the scans, and adequacy of luminal distention was evaluated on a four-point scale. Demonstration of features of various kinds of small bowel disease was analyzed. RESULTS: The taste of iso-osmotic mannitol is good (slightly sweet) and acceptable by all. Small bowel distention was excellent and moderate in most volunteers and patients. CT features of many kinds of diseases such as tumors, Crohn's disease,and small bowel obstruction, etc. were clearly displayed. CONCLUSION: Multi-detector CT enterography with iso-osmotic mannitol as negative contrast to distend the small bowel is a simple, rapid, noninvasive and effective method of evaluating small bowel disease. PMID- 15818747 TI - Effect of ZVAD-fmk on hepatocyte apoptosis after bile duct ligation in rat. AB - AIM: Retention and accumulation of toxic hydrophobic bile salts within hepatocyte may cause hepatocyte toxicity by inducing apoptosis. Apoptosis is a pathway of cell death orchestrated by a family of proteases called caspases. Z-Val-Ala-Asp (OMe)-fluoromethyl ketone (ZVAD-fmk) is a cell-permeable irreversible inhibitor of caspase. The purpose of this study was to evaluate the possible effect of ZVAD fmk on hepatocyte apoptosis after bile duct ligation in the rat. METHODS: Male Sprague-Dawley rats, weighing 250-300 g, were randomized to five groups of five rats each. Group 1 underwent common bile duct ligation and simultaneous treatment with ZVAD-fmk (dissolved in dimethylsulfoxide (DMSO)). Group 2 underwent common bile duct ligation and simultaneous treatment with Z-Phe-Ala-fluoromethyl ketone ( ZFA-fmk, dissolved in DMSO). Group 3 underwent sham operation and simultaneous treatment with the same amount of DMSO. Group 4 underwent sham operation and simultaneous treatment with the same amount of normal saline. Group 5 underwent common bile duct ligation without other manipulation. After three days, liver tissue was harv-ested for histopathologic analysis and measurements of apoptosis. RESULTS: When compared with sham operation, common bile duct ligation significantly increased hepatocyte apoptosis (P = 0.008) and ductular proliferation (P = 0.007). ZVAD-fmk significantly diminished the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation (P = 0.008 and P = 0.007, respectively). ZFA did not show the same effects. CONCLUSION: Hepatocyte apoptosis and ductular proliferation significantly increased after common bile duct ligation. ZVAD-fmk effectively diminished the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation, whereas ZFA-fmk did not. PMID- 15818748 TI - Influence of various proton pump inhibitors on intestinal metaplasia in noneradicated Helicobacter pylori patients. AB - AIM: Intestinal metaplasia (IM) is more often found in patients with Helicobacter pylori (H pylori) infection, while eradication of H pylori results in significant reduction in the severity and activity of chronic gastritis. We aimed to determine in patients with unsuccessful eradication of H pylori the role of various proton pump inhibitors (PPIs) having different mechanisms in the resolution of IM. METHODS: We confirmed endoscopically and pathohistologically (Sydney classification) the IM in 335 patients with gastritis before and after medication for eradication of H pylori (Maastricht Protocol 2002). H pylori infection was determined by using histology, urease test and culture. Control endoscopy and histology were done after 30 d and thereafter (within 1 year). Unsuccessful eradication was considered if only one of the three tests (histology, urease and culture) was negative after therapy protocol. We used omeprazole, pantoprazole, lansoprazole in therapy protocols (in combination with two antibiotics). RESULTS: We found no significant difference in resolution of IM by using different PPI between the groups of eradicated and noneradicated patients (P<0.4821 and P<0.4388, respectively). CONCLUSION: There is no significant difference in resolution of intestinal metaplasia by different proton pump inhibitors. PMID- 15818749 TI - Low gradient ascites: a seven-year course review. AB - AIM: To study the patients with low gradient ascites in hospitals of Guilan Province (northern Iran). METHODS: Patients admitted in hospitals of Guilan Province with low gradient ascites from 1993 to 2000 were enrolled in the study. Serum and ascitic fluid albumin levels were determined by biochemical reactions. The serum-ascitic albumin gradient (SAAG) less than 1.1 g/dL was considered low. Statistical analysis was performed with SPSS 9.0 software and P<0.05 was considered statistically significant. RESULTS: Of the 148 patients enrolled in the study, 72 (48.6%) were males and 76 (51.4%) were females with a mean age of 59.03+/-13.54 years. Tuberculous peritonitis was the most frequent cause of low gradient ascites in 68 (45.9%). Other most frequent causes were cancer in 62 (41.9%), nephrotic syndrome in 9 (6%), pancreatitis in 6 (4%). Peritoneal cancer was found in 22 (35%), ovarian and gastric cancers were found in 14 (22.5%) and 12 (19.3%), respectively. All of which were the causes of ascites. The mean SAAG was 0.68+/-0.19 g/dL. The mean serum and ascitic fluid albumin concentrations were higher in tuberculous patients (P<0.006), but lactate dehydrogenase (LDH) level was higher in cancer patients (P<0.0001). In peritoneal tuberculosis, mean ascitic glucose concentration was significantly lower than other patients (P<0.0001). CONCLUSION: Tuberculosis should be considered in all patients with low gradient ascites especially in developing countries (like Iran), as the first cause of ascites. In the approach to patients with low gradient ascites, ascitic fluid glucose, and LDH level are useful indicators for decision making. PMID- 15818750 TI - Oxidative stress and nitric oxide in rats with alcohol-induced acute pancreatitis. AB - AIM: Oxygen free radical mediated tissue damage is well established in pathogenesis of acute pancreatitis (AP). Whether nitric oxide (NO) plays a deleterious or a protective role is unknown. In alcohol-induced AP, we studied NO, lipooxidative damage and glutathione in pancreas, lung and circulation. METHODS: AP was induced in rats (n = 25) by injection of ethyl alcohol into the common biliary duct. A sham laparatomy was performed in controls (n = 15). After 24 h the animals were killed, blood and tissue sampling were done. RESULTS: Histopathologic evidence confirmed the development of AP. Marked changes were observed in the pulmonary tissue. Compared with controls, the AP group displayed higher values for NO metabolites in pancreas and lungs, and thiobarbituric acid reactive substances in circulation. Glutathione was lower in pancreas and in circulation. Glutathione and NO were positively correlated in pancreas and lungs of controls but negatively correlated in circulation of experimental group. In the experimental group, plasma thiobarbituric acid reactive substances were negatively correlated with pancreas thiobarbituric acid reactive substances but positively correlated with pancreas NO. CONCLUSION: NO increases in both pancreas and lungs in AP and NO contributes to the pathogenesis of AP under oxidative stress. PMID- 15818751 TI - Effects of ursodeoxycholic acid and/or low-calorie diet on steatohepatitis in rats with obesity and hyperlipidemia. AB - AIM: To evaluate the effects of ursodeoxycholic acid (UDCA) and/or low-calorie diet (LCD) on a rat model of nonalcoholic steatohepatitis (NASH). METHODS: Fifty five Sprague-Dawley rats were divided into five groups. The control group (n = 9) was fed with standard rat diet for 12 wk, NASH group (n = 10) was fed with high fat diet consisted of normal diet, 10% lard oil and 2% cholesterol for 12 wk, UDCA group (n = 10) was fed with high-fat diet supplemented with UDCA at a dose of 25 mg/(kg.d) in drinking water for 12 wk, LCD group (n = 10) was fed with high fat diet for 10 wk and then LCD for 2 wk, and UDCA+LCD group (n = 15) was fed with high-fat diet for 10 wk, followed by LCD+UDCA for 2 wk. At the end of the experiment, body weight, serum biochemical index, and hepatopathologic changes were examined. RESULTS: Compared with the control group, rats in the NASH group had significantly increased body weight, liver weight, and serum lipid and aminotransferase levels. All rats in the NASH group developed steatohepatitis, as determined by their liver histology. Compared with the NASH group, there were no significant changes in body weight, liver weight, blood biochemical index, the degree of hepatic steatosis, and histological activity index (HAI) score in the UDCA group; however, body and liver weights were significantly decreased, and the degree of steatosis was markedly improved in rats of both the LCD group and the UDCA+LCD group, but significant improvement with regard to serum lipid variables and hepatic inflammatory changes were seen only in rats of the UDCA+LCD group, and not in the LCD group. CONCLUSION: LCD might play a role in the treatment of obesity and hepatic steatosis in rats, but it exerts no significant effect on both serum lipid disorders and hepatic inflammatory changes. UDCA may enhance the therapeutic effects of LCD on steatohepatitis accompanied by obesity and hyperlipidemia. However, UDCA alone is not effective in the prevention of steatohepatitis induced by high-fat diet. PMID- 15818752 TI - c-src activating mutation analysis in Chinese patients with colorectal cancer. AB - AIM: To investigate the occurrence of cellular src (c-src) activating mutation at codon 531 in colorectal cancer patients from Chinese mainland. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay followed by sequencing and single-strand conformation polymor-phism analysis were carried out to screen 110 samples of primary colorectal cancer and 20 colorectal liver metastases. RESULTS: Only one sample showed PCR-RFLP-positive results and carried somatic codon 531 mutations. No additional mutation of c-src exon 12 was found. CONCLUSION: c-src codon 531 mutation in colorectal cancer is not the cause of c-src activation. PMID- 15818753 TI - Endoscopic findings in a patient with Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is a systemic vasculitis of the small vessels of the skin, joints, GI tract, and kidney. It preferentially affects children but may also occur in adults. We report a 60-year-old man with HSP who presented with colicky abdominal pain, bloody diarrhea, arthralgia, and skin rash. The gastrointestinal tract was viewed by upper endoscopy and colonoscopy. We found characteristic endoscopic findings in the stomach, cecum and sigmoid colon, the combination of which has rarely been demonstrated in one patient. Histologic examination of skin biopsy specimens revealed leukocytoclastic vasculitis with positive staining for IgA in the capillaries. Endoscopy appears to have substantial diagnostic utility in patients suspected of having HSP, especially when abdominal symptoms precede the cutaneous lesions. PMID- 15818754 TI - Pulmonary embolization as primary manifestation of hepatocellular carcinoma with intracardiac penetration: a case report. AB - Intracardiac manifestation of hepatocellular carcinoma (HCC) is a rare condition and an uncommon finding even at autopsy. Pulmonary tumor embolism as a presenting feature of HCC has been published only twice previously. In our case report, a 63 year-old man presented with high fever and six episodes of recurrent pneumonias during the last half year. Echocardiography was performed, a solid mass was found in the right atrium. Transesophageal echocardiography proved a tumor mass in the inferior vena cava (IVC) extending into the right atrium, abdominal ultrasound revealed tumor mass in the IVC and a solid tumor in the liver. Combined liver and heart surgery was attempted in order to remove the tumor mass from both the liver and the right atrium. Acute cor pulmonale occurred during tumor removal from the right atrium and the patient expired. In addition to local factors the possibility of embolization should arise in the background of recurrent pneumonia. Occult carcinoma must be included in possible causes of recurrent pulmonary embolism. Searching for primary malignancy should include HCC as frequent cause of hypercoagulability. In case of HCC, echocardiography is suggested because of the possibility of expansion in IVC or right atrium and tumor-embolization. PMID- 15818755 TI - Multifocal intraportal invasion of breast carcinoma diagnosed by laparoscopy assisted liver biopsy. AB - Hepar lobatum carcinomatosum (HLC) is defined as an acquired hepatic deformity consisting of an irregularly lobulated hepatic contour caused by intravascular infiltration of metastatic carcinoma. To date, only nine cases of HLC have been reported in the literature. We report a case of a 68-year-old woman showing hepatic metastasis of breast carcinoma in radiologically unidentified form. Initially, she received left partial mastectomy for breast cancer but solid hepatic metastases were identified in S(2) and S(6), 9 mo after surgery. Then, they responded to chemotherapy and radiologically disappeared. After radiological disappearance of the liver tumors, the patient's blood chemistry showed abnormal liver function. A CT scan demonstrated heterogeneous enhancement effect in the liver in the late phase, suggesting uneven hepatic blood supply. Hepatic deformity was not obvious. Laparoscopy revealed a slightly deformed liver surface with multiple indentations and shallow linear depressions. Furthermore, a wide scar was observed on the surface of S(2) possibly at the site where the metastatic tumor existed before chemotherapy. Liver biopsy from the wide scar lesion showed intraportal tumor thrombi with desmoplastic change. Because of its similarity to the histology of the original breast cancer, we concluded that the hepatic functional abnormalities and slightly deformed liver surface were derived from the circulatory disturbance caused by microscopic tumor thrombi. Besides, since the wide scar was located at the site of the pre-existing tumor, it is probable that chemotherapy was an important cause of fibrous scarring as a result of tumor regression. These morphologic findings are compatible with those of HLC. Laparoscopy-assisted liver biopsy was useful to make definite diagnosis, even though the hepatic deformity was radiologically undetectable. PMID- 15818757 TI - Metastatic low-grade endometrial stromal sarcoma of the sigmoid colon three years after hysterectomy. AB - A 49-year-old woman, who had undergone hysterectomy for low-grade endometrial stromal sarcoma (ESS) 3 years ago, presented with a 2-wk history of lower abdominal pain. Barium enema and sigmoidoscopy disclosed a polypoid submucosal tumor. Histopathologic features of biopsy specimens from the lesion were similar to those of the resected uterine ESS. Under the diagnosis of metastatic ESS of the sigmoid colon, sigmoidectomy was performed. Microscopic examination demonstrated dense proliferation of spindle cells with little nuclear atypia, which were sometimes arranged in whorled pattern around abundant arterioles. Mitotic count is below 1 in 10 high-power fields. Immunohistochemically, the neoplastic cells were strongly positive for vimentin, estrogen receptor and progesterone receptor but negative for alpha-smooth muscle actin, S-100 protein and CD34. Thus, a final diagnosis of low-grade ESS metastasis to the sigmoid colon was made. Her postoperative course was uneventful and hormonal therapy with progestational agents is entertained. PMID- 15818756 TI - Severe chronic diarrhea and weight loss in cholesteryl ester storage disease: a case report. AB - AIM: An inherited deficiency of human lysosomal acid lipase (LAL) results in the rare conditions of Wolman disease and cholesteryl ester storage disease (CESD). We want to present the rare case of CESD in an adult. METHODS: We report about an adult female patient with severe chronic diarrhea and weight loss as a consequence of CESD. Clinical examination revealed signs of malabsorption and slightly elevated liver enzymes. RESULTS: Histopathologic changes in the liver tissue and DNA sequence analysis confirmed the diagnosis of CESD due to homozygosity for the most common CESD mutation, a G934A splice site defect encoded by exon 8 of the lysosomal acid lipase (LIPA) gene. CONCLUSION: It is the first case in the literature with diarrhea as a putative symptom of CESD in adult patients. PMID- 15818758 TI - Gains for new heart disease marker. PMID- 15818759 TI - Migraine and stroke. PMID- 15818760 TI - FDA panel rules COX-2 pain drugs should remain on the market. PMID- 15818762 TI - Radioactive seeds treat prostate cancer. PMID- 15818761 TI - Common spice may slow Alzheimer's. PMID- 15818763 TI - Safety of calcium channel blockers for hypertension questioned. PMID- 15818765 TI - Common scents key to Alzheimer's diagnosis. PMID- 15818764 TI - Powerful HDL booster available OTC. PMID- 15818766 TI - Acid-blockers may increase pneumonia risk. PMID- 15818767 TI - High doses of vitamin E may increase risk of death. Talk to your doctor before taking supplements containing more than 200 IUs. PMID- 15818768 TI - I hear more and more about lumbar stenosis being a common problem with seniors. What exactly is it and what is the suggested therapy or cure if one exists? PMID- 15818769 TI - When a colon polyp is found and removed on colonoscopy, when should the next colonoscopy be done? PMID- 15818770 TI - Neurognostics question: knowledge ought to direct the hands. PMID- 15818771 TI - Post-call accidents. PMID- 15818772 TI - Post-call accidents. PMID- 15818773 TI - Post-call accidents. PMID- 15818774 TI - Coronary revascularization before vascular surgery. PMID- 15818775 TI - Coronary revascularization before vascular surgery. PMID- 15818776 TI - Immune signatures in follicular lymphoma. PMID- 15818777 TI - When doctors go to war. PMID- 15818778 TI - When doctors go to war. PMID- 15818779 TI - Acute toxicity of intravenously administered microfabricated silicon dioxide drug delivery particles in mice: preliminary findings. AB - BACKGROUND AND OBJECTIVE: Microfabricated particles with nanosized features may serve an important role in the next generation of drug delivery vehicles. Microfabrication (micro-electromechanical systems) technologies offer the promise of both structural elements (e.g., pores, reservoirs) and electromechanical features (e.g., timers, valves, actuators) built into a single particle. In order to serve as carriers to deliver drugs to systemic sites of action, such as tumors, the particles must be safe to administer intravenously. An acute safety study was performed in a mouse model, using intravenous injection of solid silicon dioxide particles created to simulate the size and shape of potential targeted drug delivery vehicles. DESIGN: Two-micron thick, square and circular, parallelepiped-shaped particles were produced with varying sizes of 2 microm, 5 microm and 10 microm using microfabrication techniques and injected into groups of mice (six mice per group) over a range of doses. End-points included acute lethality, clinical signs of toxicity and weight loss. Sections of major organs were sampled for histological examination. RESULTS: At dose levels of 1 x 10(8) particles per mouse, circular particles of 2 microm and 5 microm showed no signs of acute toxicity. Similar results were obtained with the 2 microm and 5 microm square silicon dioxide particles; however, 14-day necropsy indicates fewer 5 microm circular particles in the lung than 5 microm square particles, indicating that the shape of the particles may impact on safety. Acute lethality was observed for 10 microm particles; none of the mice injected with the 10 microm particles survived except at very low dose levels of 6 x 10(5) particles per mouse. CONCLUSIONS: Solid silicon particles greater than 5 microm in their largest dimension are cleared in the lungs and are not safe for intravenous delivery. Particles of 2-5 microm in size do not lodge predominantly in the lung and do not cause acute toxicity, but accumulate in organs such as the liver and spleen. Possible chronic toxicities associated with organ uptake of such non biodegradable particles have yet to be addressed. PMID- 15818780 TI - A bitter pill: the new Medicare prescription drug benefit. PMID- 15818781 TI - Alternative medicines need scientific grounding. PMID- 15818783 TI - Weight-loss programs don't work long term. PMID- 15818782 TI - New drug for severe pain gains approval. PMID- 15818784 TI - Ibuprofen tops acetaminophen for treating arthritis. PMID- 15818785 TI - Got hypertension? PMID- 15818786 TI - Back pain, no gain--you lose brain size. PMID- 15818787 TI - Shark cartilage cancer treatments are pseudoscience. PMID- 15818788 TI - Beta-carotene supplements pose risks. PMID- 15818789 TI - I'm a 72-year-old man, and I often wake up four or five times a night to urinate. Why is this? PMID- 15818790 TI - I've always heard that eating fish is good for my heart. Why is this so, how much should I eat during a given week, and what types are best? PMID- 15818791 TI - Produce safety action plan available. PMID- 15818792 TI - New drug to treat most common lung cancer. PMID- 15818793 TI - HHS 'Quitline' helps Americans stop smoking. PMID- 15818794 TI - Labeling changes for mifepristone. PMID- 15818795 TI - Two dietary supplements contain undeclared ingredient. PMID- 15818796 TI - New treatment for multiple sclerosis. PMID- 15818797 TI - The past and future of extant amphibians. PMID- 15818798 TI - The source of the Lisbon earthquake. PMID- 15818799 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818800 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818801 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818802 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818803 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818804 TI - Gene symbol: SEDL. Disease: spondyloephiphyseal dysplasia, late. PMID- 15818805 TI - Gene symbol: TP53. Disease: squamous cell carcinoma of head and neck. PMID- 15818806 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818807 TI - Gene symbol: SLC7A9. Disease: cystinuria, untyped. PMID- 15818809 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818810 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818811 TI - Gene symbol: GJA1. Disease: oculodentodigital dysplasia. PMID- 15818812 TI - Gene symbol: GTP cyclohydrolase I (GCH-I). Disease: dopa-responsive dystonia. PMID- 15818813 TI - Gene symbol: CYBB. Disease: X-linked chronic granulomatous disease. PMID- 15818814 TI - Gene symbol: CLN3. Disease: juvenile neuronal ceroid lipofuscinosis (Batten disease). PMID- 15818815 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818816 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818817 TI - Gene symbol: SLC7A9. Disease: cystinuria, type I. PMID- 15818818 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818819 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818820 TI - Gene symbol: SLC7A9. Disease: cystinuria, untyped. PMID- 15818822 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818823 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818824 TI - Gene symbol: SLC7A9. Disease: cystinuria, untyped. PMID- 15818825 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818828 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818829 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818831 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818832 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818833 TI - Gene symbol: NOTCH3. Disease: cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy. PMID- 15818834 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818835 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818836 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818837 TI - Gene symbol: SLC7A9. Disease: cystinuria, type non-I. PMID- 15818838 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818840 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818843 TI - Gene symbol: SLC3A1. Disease: cystinuria. PMID- 15818844 TI - [Respect for natural conditions in experimentation. Marcel Baltazard in the lineage of Charles Nicolle]. PMID- 15818845 TI - [Marcel Baltazard and typhus]. PMID- 15818846 TI - [Marcel Baltazard and rabies]. PMID- 15818847 TI - [Marcel Baltazard and the plague]. PMID- 15818848 TI - [Resistance to the plague by certain wild rodent species. I. Limitation to the epizootic process. 1963]. PMID- 15818849 TI - [B. Systematic study of a "mesofoyer" for wild animal plague in Iranian Kurdistan. 1963]. PMID- 15818851 TI - What's #1, and does it really matter? Cancer surpasses heart disease among "young" Americans, but isn't tops overall. PMID- 15818850 TI - [C. Interepizootic conservation of the plague in inveterate reservoir. Hypotheses and work. 1963]. PMID- 15818852 TI - New diet guidelines: a few steps forward, a few back. For your heart and overall health, don't follow all of the government's healthy eating recommendations. PMID- 15818853 TI - Ounces of prevention? Women--go for more! Heart disease is harder on women, making prevention efforts extra important for them at all ages. PMID- 15818854 TI - Another reason to hate Mondays. PMID- 15818855 TI - Working while sick. PMID- 15818856 TI - Ask the doctor. My doctor is focused on keeping my LDL in a good range, but she doesn't seem too concerned that my HDL is on the low side. Shouldn't it get the same attention? PMID- 15818857 TI - Neuroendoscopic observation of a large anterior communicating artery aneurysm. PMID- 15818858 TI - [Kallikrein-kinin system and organ protection]. PMID- 15818859 TI - Multiple-wave lateral shearing interferometry for wave-front sensing. AB - Multiple-wave achromatic interferometric techniques are used to measure, with high accuracy and high transverse resolution, wave fronts of polychromatic light sources. The wave fronts to be measured are replicated by a diffraction grating into several copies interfering together, leading to an interference pattern. A CCD detector located in the vicinity of the grating records this interference pattern. Some of these wave-front sensors are able to resolve wave-front spatial frequencies 3 to 4 times higher than a conventional Shack-Hartmann technique using an equivalent CCD detector. Its dynamic is also much higher, 2 to 3 orders of magnitude. PMID- 15818860 TI - Improved retrievals of the optical properties of cirrus clouds by a combination of lidar methods. AB - We focus on improvement of the retrieval of optical properties of cirrus clouds by combining two lidar methods. We retrieve the cloud's optical depth by using independently the molecular backscattering profile below and above the cloud [molecular integration (MI) method] and the backscattering profile inside the cloud with an a priori effective lidar ratio [particle integration (PI) method]. When the MI method is reliable, the combined MI-PI method allows us to retrieve the optimal effective lidar ratio. We compare these results with Raman lidar retrievals. We then use the derived optimal effective lidar ratio for retrieval with the PI method for situations in which the MI method cannot be applied. PMID- 15818861 TI - [Pseudolymphoma of the breast nipple. The problem overview]. AB - Clinical, pathological, and molecular-genetic features as well as etiology of cutaneous pseudolymphoma (CPL, cutaneous lymphoid hyperplasia, lymphocytoma cutis) of the breast nipple are summarized. CPL presents as a nipple induration and it is often suspected to be Paget carcinoma pre-operatively. Histologically, atypical microscopic features of a dense lymhoid infiltrate with follicles often mislead to the diagnosis of a malignant lymphoma. However, CPL runs a benign course. Rare cases of CPL contain a clonal lymphoid population. A substantial number of CPL in the breast nipple is caused by antigenic stimulation by Borrelia burgdorferi. In some patients a tick bite is documented. CPL of the breast is commonly treated by excision, but some patients may be cured by antibiotic therapy. The presence of Borrelia burgdorferi should be detected using methods of serology, culture, and molecular biology. Beside CPL, the differential diagnosis of the breast nipple lesions further includes Paget carcinoma, eczema, and florid papillomatosis. PMID- 15818862 TI - [Virtual colonoscopy as a part of the algorithm of the facultative examinations prior to the colorectal carcinoma procedure]. AB - The authors present their own initial experience with virtual colonoscopy. Both benign and malignant stenosing processes of the colon and rectum, which make examining oral parts of the colon using colonoscopy or irrigography impossible, are the main indication for the procedure described. The authors' first experience with this method is totally positive. The more experienced the examiners, the better interpretation of the findings not only by radiologists but also by operating surgeons. In certain indications, the examination has its firm place in the algorithm of the facultative examinations prior to the colorectal carcinoma surgical procedures. PMID- 15818863 TI - Three-dimensional localization and optical imaging of objects in turbid media with independent component analysis. AB - A new approach for optical imaging and localization of objects in turbid media that makes use of the independent component analysis (ICA) from information theory is demonstrated. Experimental arrangement realizes a multisource illumination of a turbid medium with embedded objects and a multidetector acquisition of transmitted light on the medium boundary. The resulting spatial diversity and multiple angular observations provide robust data for three dimensional localization and characterization of absorbing and scattering inhomogeneities embedded in a turbid medium. ICA of the perturbations in the spatial intensity distribution on the medium boundary sorts out the embedded objects, and their locations are obtained from Green's function analysis based on any appropriate light propagation model. Imaging experiments were carried out on two highly scattering samples of thickness approximately 50 times the transport mean-free path of the respective medium. One turbid medium had two embedded absorptive objects, and the other had four scattering objects. An independent component separation of the signal, in conjunction with diffusive photon migration theory, was used to locate the embedded inhomogeneities. In both cases, improved lateral and axial localizations of the objects over the result obtained by use of common photon migration reconstruction algorithms were achieved. The approach is applicable to different medium geometries, can be used with any suitable photon propagation model, and is amenable to near-real-time imaging applications. PMID- 15818864 TI - A comparative reliability analysis of computer-generated bitemark overlays. AB - This study compared the reliability of two methods used to produce computer generated bitemark overlays with Adobe Photoshop (Adobe Systems Inc., San Jose, CA). Scanned images of twelve dental casts were sent to 30 examiners with different experience levels. Examiners were instructed to produce an overlay for each cast image based on the instructions provided for the two techniques. Measurements of the area and the x-y coordinate position of the biting edges of the anterior teeth were obtained using Scion Image software program (Scion Corporation, Frederick, MD) for each overlay. The inter- and intra-reliability assessment of the measurements was performed using an analysis of variance and calculation of reliability coefficients. The assessment of the area measurements showed significant variances seen in the examiner variable for both techniques resulting in low reliability coefficients. Conversely, the results for the positional measurements showed no significant differences in the variances between examiners with exceptionally high reliability coefficients. It was concluded that both techniques were reliable methods to produce bitemark overlays in assessing tooth position. PMID- 15818865 TI - Author responds to letters on "Children in clinical research: a conflict of moral values" (AJOB 3:1). PMID- 15818866 TI - Author responds to "Review of Carl Elliott, Better than well: American medicine meets the American dream" by Paul Root Wolpe (AJOB 3:3). PMID- 15818867 TI - [Japanese translation of common terminology criteria for adverse events (CTCAE), and instructions and guidelines]. PMID- 15818868 TI - Urinary incontinence in prostate cancer patients treated with external beam radiotherapy. AB - BACKGROUND AND PURPOSE: To describe the incidence of urinary incontinence among prostate cancer patients treated with external beam radiotherapy (RT) and to investigate associated risk factors. PATIENTS AND METHODS: One thousand and hundred ninety-two patients with >or=24 months follow-up were the subjects of this series. All patients received between 50 and 72 Gy in 20-37 fractions (median 66 Gy/33#). Post-RT urinary incontinence was scored by direct patient interviewing according to the modified RTOG/SOMA scale: Grade 1--occasional use of incontinence pads, Grade 2--intermittent use of incontinence pads, Grade 3- persistent use of incontinence pads, and Grade 4--permanent catheter. Risk factors investigated were: age, diabetes, TURP prior to RT, elapsed time from TURP to RT, clinical stage, RT dose and presence of Grade >or=2 acute GU and GI toxicity. Non-parametric, actuarial univariated (Kaplan-Meier) and multivariated tests (MVA, Cox regression) were performed. RESULTS: Median follow-up for the group is 52 months (24-109). Thirty-four patients (2.9%) had incontinence prior to RT, which was more common in TURP patients (7.8% vs 1.6% P<0.001). These are excluded from further analysis. Fifty-seven patients (4.9%) developed Grade 1 incontinence, 7 (0.6%) Grade 2, and 7 (0.6%) Grade 3. There was no Grade 4 incontinence. Actuarial rates for Grade >or=1 and >or=2 incontinence at 5 years are 7 and 1.7%, respectively. Risk factors on MVA associated with the development of Grade 1 or worse incontinence are pre-RT TURP (5-year rates 10% vs 6%, P=0.026), presence of Grade >or=2 acute GU toxicity (5-year rates 11% vs 5%, P=0.002). Age, diabetes, clinical stage, elapsed time from TURP to RT, RT dose or fraction size, acute GI toxicity were not significant. Patients who underwent post-RT TURP or dilatation for obstructive symptoms (4.3%), were more likely to develop Grade 2-3 incontinence (5-year rate 8 vs 1.5%, P=0.0015). CONCLUSIONS: Grade 2 or greater urinary incontinence is rare among patients who have been treated with external beam radiotherapy. Associated risk factors are pre-RT TURP and the presence of increased acute GU toxicity. Post-radiaton TURP increases the risk of incontinence five-fold. PMID- 15818869 TI - Aneurysm check urged for some men. For older smokers, a quick test for a bulging aorta could save lives. PMID- 15818871 TI - [Measles and medical care of the Edo era]. AB - In Bunkyu 2, measles was prevelant very much in all over Japan and on that occasion resembles it, and a measles picture and a called ukiyoe print are published in large quantities in a town of Edo, and taboo of measles and medical care information such as prescription of medicine were pictured in a measles picture, but, by a conventional study, they are considered to depend on private enterprise medical care and a supersitition, the medicine book that it is easy. Because outlook on authority of assumed the history of measles medical care before reaching it in Bunkyu 2 it, this report lectured on intellect of medicine and medical care showing what they developed extensively in people level to the measles picture for this. PMID- 15818870 TI - The involvement of professional medical writers in medical publications: results of a Delphi study. AB - OBJECTIVE: Using a Delphi Consultation process, a group of medical writers established by the European Medical Writers Association (EMWA) set out to determine the current thinking on the problems of ghostwriting in medical publications and what should be done about them. In this context, ghostwriting is where a professional medical writer prepares a manuscript on behalf of a named author, but the writer is not listed as an author. METHODS: A 4-round Delphi consultation process was conducted via email to generate statements about the main issues in ghostwriting. Participants rated their agreement with the statements on a scale of 0-10. RESULTS AND CONCLUSIONS: Members of the task force strongly believed that professional medical writers can improve the quality of scientific papers, but that fact is often not recognised outside the medical writing profession. At least in part, this is because of a perception that ghostwritten papers may have been inappropriately influenced by pharmaceutical companies. One theme that emerged strongly from the discussions was transparency. Members thought it very important that the existence of a ghostwriter should always be made clear to the reader. Another strong theme was the importance of defining in detail what practices relating to ghostwriting are ethical, and what practices are not. This definition of ethical ghostwriting should be widely known, and unethical ghostwriting should be strongly condemned. Use of the term 'ghostwriting' itself was questioned. Members of the task force felt that use of a more neutral term should be encouraged. The task force suggested various activities for ensuring that above the objectives could be met, including discussions with other interested parties, such as journal editors and pharmaceutical companies, educating medical writers about ethical practices, further research into ghostwriting, and developing guidelines for ethical medical writing. PMID- 15818872 TI - [Questions relating to Manase Gensaku's medical writings. Did Manase Gensaku write "Sankyoshiyo Bassui" and "Saiminki"?]. AB - This study investigated the texts attributed to Manase Gensaku (1549-1631, the second Mamase Dosan), successor to the first Manase Dosan (1507-1594). The investigation yielded the following results:(1) "Saiminki," "Sankyoshiyo Bassui" and "Iden Fukyuroku," said to have been authored by Manase Gensaku, constitute the first Manase Dosan's works. Manase Gensaku is thought to have revised "Saiminki," recompiled "Sankyoshiyo Bassui," and published "Iden Fukyuroku."(2) Manase Gensaku's medical writings, "Denshin Hoho," "Keitokuho" and "Jozanho," have the same contents.(3) Late in his life, Manase Gensaku wrote the pharmaceutical volumes, "Kyushin Zappo," "Iho Joboku" and "Iho Kenko." These books described pharmaceuticals that were described by Manase Dosan, as well as pharmaceuticals that were being developed at that time.(4) "Iho Meikan," the most important work of Manase Gensaku, contains descriptions of almost all of the pharmaceuticals described by the first Manase Dosan.(5) The principal aim of Manase Gensaku was the production of straightforward, compact and convenient medical texts that could be used by clinicians. This was because most of Manase Dosan's medical texts were too extensive to be useful for practicing clinicians of the time.(6) In addition to working on aspects of internal medicine investigated by Manase Dosan, Manase Gensaku made radical improvements in disciplines such as materia medica, regimens, acupuncture and moxibustion. PMID- 15818873 TI - [Transition of the doctrine from ancient to medieval China for embryo and fetus on each month during pregnancy period]. AB - There are written some doctrines about growth image of embryo and fetus, hygiene for woman on each month during pregnancy period etc. in ancient Chinese medical book the Taichan Shu, which compiled over 2200 years ago. Following results were obtained by a study on the transition doctrines from that of Taichan Shu to those of medieval Chinese and Japanese medical books such as the Zhubing Yuanhou Lun, the Qianjin Fang and the Ishin Po. (1) Doctrines about growth and hygiene on embryo and fetus had been developed on each coming book from the contents of the Taichan Shu descent. Doctrines about hygiene were mainly for pregnant woman, and some methods of them were told by using figure image and the five elements theory in the part. (2) There could be found additional doctrines about meridian and internal organs theory in the Zhubing Yuanhou Lun, the Qianjin Fang and the Ishin Po. And those additional doctrines were made under the influence of the Maijing and the Suwen. New doctrines about hygiene for pregnant woman found in the Qianjin Fang and the Ishin Po also have relationships with meridian and internal organs theory. (3) Growing organs of embryo and fetus described in the Zhubing Yuanhou Lun and the Qianjin Fang correspond with the internal organs theory. Purpose of those descriptions is estimated to be equipped much more organs for embryo and fetus than those of the Taichan Chu. PMID- 15818875 TI - [Andreas Vesalius in the Spanish Court]. AB - After the publication of "Fabrica," Andreas Vesalius entered the Spanish court and became a court physician to Charles the Fifth, Holy Roman Emperor, and then to Philip the Second, Spanish king. The author studied this process and its historical background. The ancestors of Vesalius had close relations with the Hapsburgs and the dukes of BUrgundy, and served them as court physician or a court pharmacist. Vesalius was born in Brussels, obtained his degree at the University of Padua, Italy, became professor of anatomy and surgery there, and published "Tabulae Anatomicae Sex" and "Fabrica."In the ear of the Spanish court, the treatments of Henry the Second, French king, and of Don Carlos, Spanish crown prince, are famous among Vesalius' medical contributions. In the year of his resignation, Charles the Fifth conferred the title of count palatine on Vesalius. PMID- 15818874 TI - Death anxiety in clinical and non-clinical groups. AB - The Arabic Scale of Death Anxiety (ASDA) was administered, individually, to 7 groups (N = 765) of Egyptian normal participants (non-clinical), anxiety disorder patients, and patients suffering from schizophrenia (males and females), and addicts (males only). They were generally matched as groups according to age, occupation, and education. The female and male anxiety disorder patients means were, respectively, significantly higher than the means of the other 5 groups on the ASDA, while male schizophrenics attained the lowest mean score in proportion to all the other 6 groups, including the non-clinical 2 groups. All female groups have higher mean scores than their male counterparts. PMID- 15818876 TI - [The documents of 1794 medical examination at Igakkan]. PMID- 15818877 TI - Martin David Kamen (27 August 1913 - 31 August 2002). PMID- 15818878 TI - Robert K. Merton (4 July 1910 - 23 February 2003). AB - Robert K. Merton, one of the greatest sociologists of our time, and the doyen of the sociology of science, died in Manhattan on 23 February 2003, at the age of ninety-two. He was an exemplary discipline-builder who formulated key concepts with which to perceive and solve sociological problems, a masterful teacher, and a kind colleague. His passing left a large void in the intellectual world of social science. PMID- 15818879 TI - Trumpeting racial differences among healthcare providers is harmful. PMID- 15818880 TI - Political and financial agendas now set the "norm" for Standards of Care. PMID- 15818881 TI - Unusual nerve-artery relationship in microvascular exploration for tic douloureux. PMID- 15818882 TI - Results of multimodality treatment for 141 patients with brain arteriovenous malformations and seizures: factors associated with seizure incidence and seizure outcomes. PMID- 15818883 TI - Death after late failure of endoscopic third ventriculostomy: a potential solution. PMID- 15818884 TI - Rate of resolution of histologically verified intracranial tuberculomas. PMID- 15818885 TI - Stimulation of the subthalamic nucleus in Parkinson's disease does not produce striatal dopamine release. PMID- 15818886 TI - Detection of intracranial aneurysms with two-dimensional and three-dimensional multislice helical computed tomographic angiography. PMID- 15818887 TI - Depression in patients with high-grade glioma: results of the Glioma Project. PMID- 15818889 TI - Predicting function. PMID- 15818888 TI - Grading of subarachnoid hemorrhage: modification of the World Federation of Neurosurgical Societies scale on the basis of data for a large series of patients. PMID- 15818891 TI - On unhealthy pleasures. PMID- 15818890 TI - Evolution of the brain: from behavior to consciousness in 3.4 billion years. PMID- 15818892 TI - Livelihood hazards. PMID- 15818893 TI - Using live FRET imaging to reveal early protein-protein interactions during T cell activation. AB - The emerging challenge for proteomics in general and lymphocyte biology in particular is to understand protein-protein interactions in the dynamic context of the living cell. Particularly interesting are the molecular dynamics of the T cell receptor-CD3 complex and other immunoreceptors in immune synapses. Fluorescence (or Forster) resonance energy transfer (FRET) is one of the few techniques that are capable of giving dynamic information about the nanometer range proximity between molecules, as opposed to simply the subcellular co localization that is provided by fluorescence microscopy. Spectral changes in fluorescence intensity and down modulation of donor lifetime are the basis for rapidly developing approaches to real-time FRET imaging. With two-photon excitation, FRET can now be extended to in vivo imaging. PMID- 15818895 TI - Great contributions of E. Donnall Thomas to the development of clinical applications of bone marrow transplantation, leading to the 1990 Nobel Prize in Medicine/Physiology. PMID- 15818898 TI - [Association between diversity in the Src homology 2 domain-containing tyrosine phosphatase binding site of Helicobacter pylori CagA protein and gastric atrophy and cancer. J Infect Dis 2004;189:820-827]. PMID- 15818899 TI - [Spectroscopic study of the 1pig Rydberg states of Na2]. AB - Six highly excited 1pig states of the Na2 molecule have been investigated by optical-optical double resonance (OODR) flourescence excitation spectroscopy. Absolute vibrational numberings are determined and molecular constants and RKR potential curves are obtained. The lowest 10 1pig states have been relabeled according to the hydrogenic nllambda characters of the Rydberg orbital and classified into core-penetrating and core-nonpenetrating Rydberg states. PMID- 15818900 TI - [Observation of autoionization levels in uranium I]. AB - A number of Rydberg and autoionization levels of U I have been studied using three-step resonant ionization methods with three pulsed tunable dye lasers. Energy levels of uranium atom have been measured, which were located in the 49898 50880 cm(-1) energy interval. PMID- 15818901 TI - [Gas-sensing transmittivity spectrum of two-dimension SnO2: TiO2 compound film in ammonia vapor]. AB - In this paper, we studied the gas-sensing transmittivity spectrum of the two dimension SnO2: TiO2 compound film in ammonia vapor. It was observed that when the gas concentration was relatively low the transmittivity decreased as the gas concentration decreased, and when the gas concentration was relatively high, the transmittivity increased as the gas concentration increased. We considered the effect of the wet vapor, and explained this phenomenon. PMID- 15818902 TI - [The application of ion detection in the study of spectroscopy of Na2]. AB - Ion detection technique has been successfully used in the study of Rydberg states of Na2 by pulse optical-optical double resonance. High-lying Rydberg states near ionization limit have been observed and high sensitive and high gain spectra have been obtained. This technique is very useful for molecular spectroscopy. PMID- 15818903 TI - [Photoacoustic spectroscopy investigation of intramolecular energy transfer in Eu beta-disketones]. AB - The complex crystals of Eu(III) with AA, HFA and TTA have been synthesized. The absorption intensities of Eu-AA, Eu-HFA and Eu-TTA are different while the photoacoustic (PA) signals of the corresponding solid complexes are almost the same, which shows that the probabilities for nonradiative transition of these chelates are different. The results of the fluorescence spectra are identical. The PA signal was obtained by detecting the heat generated through the nonradiative processes after absorption, and the intramolecular energy transfer processes of Eu-beta-diketones were studied from a new point of view. PMID- 15818904 TI - [Trace analysis of Zn-doped compound semiconductor material by resonant ionization mass spectrometry]. AB - An experimental setup and the procedure for the laser resonant ionization mass spectrometry (RIMS) have been described. Both an optical spectrum and a mass spectum have been shown. The detection limit that can be reached by using this procedure has been estimated. PMID- 15818905 TI - [Primary study on damage effects of low energy ions on tyrosine aqueous solution]. AB - Low energy ions were produced in gas arc discharge, and the ions were accelerated into tyrosine (Tyr) aqueous solution. Changs in UV and infrared spectra of Tyr after this kind of ions' action were investigated. It was showed that the ions caused various damages on Tyr molecules. The ions not only destroyed the molecules, but also synthesize were new chemical groups with elements in the solution, then the new groups were inserted into the damage fragments to generate new substances. This is called "ions deposition" damage effects of low energy ions on substances. PMID- 15818906 TI - [Photocurrent spectra of undoped LECSIGaAs]. AB - Extrinsic photocurrent spectra of undoped LECSIGaAs have been measured to investigate the deep levels in this material. A broad photocurrent response band M1 ranging from 0.40 to 0.70 eV has been observed. M1 band reveals five peaks at 0.46, 0.49, 0.56, 0.65 and 0.69 eV, respectively. The onset thresholds for the 0.46, 0.49, 0.56 and 0.69 eV peaks are 0.44, 0.47, 0.51 and 0.67 eV, respectively. It is likely that M1 band is due to different deep levels and each of these deep levels has a concentration larger than 10(14) cm(-3). The origin of M1 band is discussed and possible associations of the 0.44, 0.47 and 0.51 eV thresholds with copper acceptor, EL3 and oxygen donor are presented. The relationship between M1 band and the photoresponse behavior of drain current of GaAs field effect transistor was also investigated. PMID- 15818907 TI - [Spectroscopic assay, chemical modification and activity of three kinds of polyoses]. AB - Infrared spectra and nuclear magnetic resonance spectra of chitosan, pachyman and amorphalls mannan were studied. The difference among the infrared spectra of chitosan under different conditions was disscused. The modification of Methylolation of the three kinds of polyoses was made and the relationship between the chemical modification and the antitumor activity was also studied. PMID- 15818908 TI - [Joint rotational analysis of twenty six bands of the H2(80) Se molecule]. AB - This paper expressed the theory of Global fit model. Global fit of all known up today rotation-vibration energies of twenty four states of the H2(80) Se molecule had been made, and the corresponding set of 112 parameters, which reproduce the initial 3516 upper energies with the mean accuracy of 10.6 x 10(-3) cm(-1), were obtained. On this base fine rotational structures of the local-mode bands (500, A1) and (401, B2) were analyzed with 252 upper energies, which were analyzed in the normal-molde and local-mode also. In the end, with the results above, joint rotation-vibration energies of twenty six states of H2(80) Se molecule were made, and the corresponding set of 114 parameters obtained, which reproduce the initial 3768 upper energies with the mean accuracy of 9.62 x 10(-3) cm(-1). PMID- 15818909 TI - [The substituent structures and characteristic infrared spectra of alpha-furan esters]. AB - The characteristic infrared bands of the alpha-furoic esters, alpha-furancarbinol esters and alpha-furanacrylic esters were assigned, and the changing rules of the IR absorption frequency of the alpha-furan esters with alpha-substituent structure were discussed. The results indicated there are three characteristic bands in alpha-furan ring. The strong absorption at 1641 cm(-1) is assigned to the C=C double bond stretching mode in the alpha-furanacrylic esters, while the absorption at 973 cm(-1) is the out-of-plane =C-H bending mode for a trans di substituted alkene, the strong absorption at about 1713 cm(-1) is assigned to a C=O stretch, and the three strong absorptions near 1305, 1260 and 1165 cm(-1) are due to the symmetric and assymmetric stretching vibrations of C-O-C bond in the ester groups. PMID- 15818910 TI - [UV-Vis spectroscopic characterization of LB films of hydroxyl-substituted porphyrin derivatives]. AB - The LB films forming properties of two hydroxyl-substituted porphyrins with double hexadecyl chains have been characterized. UV-Vis spectra show that the two porphyrins possess various aggregation behaviors in LB films. The porphyrin with symmetrically attached chains tend to form J-aggregate and the other asymmetric porphyrin can form hydrogen-bonded aggregate in LB films. The orientation of porphyrin rings was examined by polarized UV-Vis spectra. The results indicate that the situation of side-chain attached on porphyrin ring has no influence on porphyrin ring orientation, but has great influence on mean molecular areas or the distanses between porphyrin rings, and so as to affect the aggregation behaviors of porphyrin in LB films. PMID- 15818911 TI - [The visible spectra study of the conformation equilibrium of iron-metal-free porphyrin dimers]. AB - A series of iron(III)-metal-porphyrin dimers linked with flexible alkoxyl chain at para position of two phenyl rings have been synthesized and characterized. An equilibrium of two conformations of porphyrin dimers (open and closed) has been studied by visible spectra. The intensity of absorption band 680 nm relates to equilibrium of two conformations. With the increase of the length of the alkoxyl chain,the closed conformation forms more easily. With the addition of axial ligand, the ferric ion of the dimers tends to bind with imidazole, and the intensity of the band 680 nm gradually decreases and disappears. PMID- 15818912 TI - [The study on the structure of 5,10,15,20-tetra-(4 hexadecylpyridiniumyl)porphyrin LB films]. AB - The film-forming properties of 5,10,15,20-tetra-(4 hexadecylpyridiniumyl)porphyrin bromide (TC16PyP) on the air/water interface were studied. LB films of the amphiphilic porphyrin were prepared on the glass, quartz and SnO2 OTE substrates. The morphology and structure of TC16PyP LB films were characterized by scanning tunnelling microscope, UV-Vis absorption spectra, fluorescence spectra and low-angle X-ray diffraction. The experimental results indicated that the amphiphilic porphyrin TC16PyP has good film-forming properties on the air/water interface, its LB films are stable, in TC16PyP monolayer or LB films the long alkyl chains are not straight and the porphyrin macroring is lying nearly parallel to the substrate surface. PMID- 15818913 TI - [Analysis of Cu-O bond stretching vibration in CuO2 plane of Bi-system high temperature superconductor]. AB - Infrared absorption spectra of Bi-system high zero resistance temperature (Tc) superconductors were measured. The relation between Cu-O bond stretching vibration in CuO2 plane and superconducting properties of Bi-system high Tc superconductors were analysed and discussed. The result of analysis indicated that the change of absorption peak strength of Cu-O bond stretching vibration was similar to the change of Tc. The reason was related to the coupling between the phonon and free carrier. And according to the theory of molecule vibration, the frequencies of Cu-O bond stretching vibration in CuO2 plane were calculated on the base of simplified model. PMID- 15818914 TI - [Studies on decomposed crop residues by FTIR differential analysis]. AB - The decomposing process of corn leaf residues (CLR) was studied by FTIR differential analysis, and the differential spectra were compared with normal spectra. The result showed that the purification process to remove inorganic matters from decomposed CLR could be omitted when differential analysis is used, and the differential spectra were clear and distinct. As far as the studies of decomposed crop residues, the FTIR differential analysis was a convenient and forthright method. PMID- 15818915 TI - [Omega-mercaptohexanoic acid self-assemble-modified gold electrode and its catalytic property for redox reaction of chlorophylls]. AB - Omega-mercaptohexanoic acid (6-MHA) self-assemble-modified gold electrode has been made by the authors. The SERS spectrum and the variations of double-layer capacitance (Cd) which occurred in the formation of 6-MHA self-assembled monolayers (SAM) on gold electrode surface were studied. The structure model of this SAM was deduced. The results show that 6-MHA can form stable SAM on gold electrode, and this 6-MHA SAM electrode has obvious catalysis for the redox reaction of chlorophylls. PMID- 15818916 TI - [Determination of 14 rare earth impurities in Y2O3 by petroleum sulfoxide extraction chromatography]. AB - Petroleum sulfoxide-NH4SCN extraction chromatography was developed for the separation of Y from other rare earth elements. Some chromatographic parameters were chosen and separation factors between Y and other rare earth elements were determined. A column of resin coated with petroleum sulfoxide was used. The Y in the sample was eluted with NH4SCN, then titrated with EDTA. The recovery was 67% 120% and relative standard deviation +/- 4.0%-19.4%. This method can be applied to the determination of trace amounts of rare earth impurities in Y2O3 with a purity of 99.9999%-99.99999%. PMID- 15818917 TI - [Determination of rare-earth impurities in ultra-highly pure Er2O3 and Ho2O3 by [HEH(HEP)] extraction chromatography-atomic emission spectrometry]. AB - This article applied [HEH(HEP)] (2-ethyhexyldrogen-2-ethylhexyl phosphonate) extraction chromatography to separate 14 rare earth impurities from ultra-highly pure Er2O3 and Ho2O3, and then the impurities were determined by atomic emission spectrometry. The average percentage recovery for each element is in the range of 70%-140%. The relative standard deviations of the determination are +/- 3.3% 2.2%. This method can be applied to the determination of the trace amounts of rare eath impurities in Er2O3 and Ho2O3 with a purity of 99.9999%-99.99999%. PMID- 15818918 TI - [Determination of trace aluminium by ETV-ICP-AES]. AB - The effect of mixed halogenation agent (polytrifluorochloroethylene (PTFCE)/hexachloroethane (C2Cl6)) on the vaporization of aluminium has been studied in electrothermal vaporization inductively coupled plasma atomic emission spectrometry (ETV-ICP-AES). The mixed halogenation agent hastens the vaporization of Al to the plasma at low temperature compared with individual halogenation agent, and the metallic material can be prevented affectively from forming in the furnace. The detection limit of this method was 0.12 ng with RSD of 3.8% (Al: 0.3 microg x mL(-1), n = 7). The analytical results of real samples were in good agreement with those of pheumatic nebulization (PN)-ICP-AES, and the recoveries were in the range of 94.1%-102.5%. PMID- 15818919 TI - [Multi-channel probing for emission spectra of metals stimulated by LD pumping laser]. AB - Emission spectra of several metals stimulated at a tiny spot by an YAG laser pumped by a LD were obtained analyzed by means of a CCD optical multi-channel analyzer. The emission spectra collected and analyzed by multi-channel with high signal to noise ratio and high dynamic range were obtained, and the wavelengths accord well with the standard spectra of these metals. Application of this method was discussed. PMID- 15818920 TI - [Study on the mutual interference of rare earth elements by ICP-AES]. AB - In this paper, the mutual interference of rare earth elements have been studied by ICP-AES. Thirty seven analyte lines of sixteen rare earth elemnts have been investigated. It is found useful to correctly select analyte lines of rare earth elements and overcome their mutual interference. PMID- 15818921 TI - [Determination of gold-beryllium alloy by inductively coupled plasma-atomic emission spectrometry]. AB - An ICP-AES method was established for the determination of Be and impurity elements in Au-Be Alloy, such as platinum, palladium, rhodium, iridium, silver etc. The interference was corrected by using the equivalant concentration technique. The detection limit in 2.5 g sample varied from 5 x 10(-5) to 2 x 10( 4%). The ralative error for Be was less than 2%. PMID- 15818922 TI - [Study of the background absorption of ErCl3 and Er(NO3) matrices in GFAAS]. AB - The investigation shows that the background absorption of Er matrices is significantly wavelength dependent. The effects of ashing temperature, heating mode, atomization temperature, recording time and matrix modifiers on the non specific absorption of Er matrices have also been studied. PMID- 15818923 TI - [Study on application of crosslinked chitosan to the speciation analysis of selenium]. AB - The adsorption of Se(IV)/Se(VI) by crosslinked chitosan was studied and the adsorption mechanism was concerned. The results indicated that the adsorption of Se(VI) was 95% at pH=4, while almost no adsorption for Se(IV). The adsorbed Se(VI) was quantitatively desorbed with 1 mol/L HCl after adsorption of Se(VI). A new method for the determination of trace Se(IV)/Se(VI) and organic selenium in environmental samples with hydride generation and atomic absorption spectphotometry has been developed. The detection limit of this method was 20 ng/L, the recovery was 93%-106% and the coefficient of variation was 2.0%-5.0%. PMID- 15818924 TI - [Determination of trace chromium and nickel in human hair using FAAS with flow injection extraction system]. AB - Trace amount of chromium and nickel was determined using a flow injection on-line extraction FAAS system. The measurement sensitivity for Cr and Ni increased by a factor of 20.1 and 14.3 respectively. For the national standand human hair sample and peach leafage sample, the measurement results are satisfactory. PMID- 15818925 TI - [Determination of trace arsenic using GFAAS with metal-coated graphite tube]. AB - The sensitivity, precision and disturbance-resisting property were studied by using metal-coated graphite tube for electrically heated atomic absorption spectrometry. The surface characteristic of the metal-coated graphite tube was studied by using X-ray diffraction meter and scanning electron microscope. The mechanism of satisfactory performance was discussed. PMID- 15818926 TI - [Simultaneous determination of europium and copper in rare earth oxide by use AAS PLS method]. AB - Partial least squares regression was used to compensate for spectral "overlap" interference of Eu 324. 753 nm with Cu 324.754 nm in atomic absorption spectrometry. We could only use the copper element hollow-cathode lamp to simultaneous determine Eu and Cu in synthetic samples and rare earth oxide, and obtained satisfactory results. PMID- 15818927 TI - [Determination of trace zinc in milk using pulse nebulization atomic absorption spectrophotometry technique]. AB - A rapid, diect, pulse nebulization method for the determination of trace zinc in milk by FAAS with model WYX-402A atomic absorption spectrometer is described. The influence of pulse nebulization volume, uptake rate of solution and other factors on the uptake signal has been discussed. The milk sample was diluted with 1.5% HCl solution. A 100 microL aliquot of the solution was then nebulied and the peak height of the transient output pulse was monitored. The result of the determination of the 1+4 diluted sample is in agreement with that of digested sample with HNO3-HClO4. The average recovery of 99.0% with relative standard deviation of 3.3% was achieved and the detection limit was 0.028 microg/mL. The method is simple, rapid, accurate and precise. PMID- 15818928 TI - [Principle and application of scattering power function method in X-ray fluorescence analysis. II. Incoherent scattering radiation]. AB - It was discovered that the relationship between mass attenuation coefficient and incoherent scattering is not an inverse proportion but a power function in a wide range of matrix compositon. Experimental results are in good agreement with this theoretical predition. A new method is recommended for matrix absorption correction according to the discovery and used for the determination of microelement Sr in geological samples as an application example. The accuracy of the results of heavy absorption samples obtained by this method is much better than those obtained by the traditional scattering internal method. PMID- 15818929 TI - [A simple method for the determination of tungsten ores by XRF spectrometer]. AB - A method for the determination of the major elements in tungsten ore using X-ray fluorescence spectrometer (PW 1404) was proposed. Tungsten ore and pure WO3 were fused, of which glass disc was made with the mixing flux of both Li2B2O7 and Na2B2O7. WO3 (%) was calculated by the relative intensity of the analytical line. Satisfactory analytical results were obtained. PMID- 15818930 TI - [The mechanism of absorbance increase and decrease in liquid-core optical fiber spectrophotometry]. AB - In this paper, the mechanism of absorbance increase and decrease in liquid-core optical fiber spectrophotometry is studied based on optical fiber theory and the results demonstrate that the absorbance increase and decrease result from the light scatter and propagation in optical fiber and depend on the scatter of samples and solvents. The experimental results are given. PMID- 15818931 TI - [Study on the surface diffuse reflectance ultraviolet-visible spectra of the multicomponent metal catalysts]. AB - In this paper, the surface diffuse reflectance ultraviolet-visible spectra (DRUVS) of two and three components supported metal catalysts for methanation of CO2 by hydrogenation were studied, and the relationship between the activity of catalytic-hydrogenation and the interaction among the components of the catalysts was revealed. Based on the analysis of the DRUVS of the catalysts with defferent amount of promoters in two component catalysts system, the dependence relationship between the characteristic strength of the DRUVS and the catalytic activity was worked out. PMID- 15818932 TI - [Flotation spectrophotometric determination of copper on isochromatic dye ion pair with crystal violet and bromopnenol blue]. AB - Bromophenol blue (BPB) was produced and entered into the aqueous phase when NaOH solution of pH = 10 was added to Cu(biq)2BPB by trichloromethane and isoamyl alcohol (20:1) extractive. And then CV x BPB was floated by crystal violet (CV) with benzene solution. The flotation was dissolved in ethanol and the absorbance of the solution measured at 590 nm. The sensitivity was raised because of the two dyes assistant effect. The molar absorptivity was 1.45 x 10(5) L x mol(-1) x cm( 1). Copper in the sample was separated first by extracting the Cu(biq)2BPB complex with trichloromethane and isoamyl alcohol, thus achieving a high selectivity. Beer's law was obeyd in the range of 0-0. 4 mg/L with a relative standard deviation of 3.6%. For 4.8 x 10(-8) g/mL copper solution, the recoveries were 97.8%-101.7%. PMID- 15818933 TI - [Luminescence characteristics of simple fluorides containing Ce3+ and Tb3+ ions]. AB - A series of phosphors containing Ce3+, Tb3+ ions in CaF2 and AlF3 were synthesized in a solution reaction and solid reaction method. Their excitation and emission spectra were determined. The results indicated that Ce3+ could not effectively transfer energy to Tb3+ in matrix CaF2 and AlF3 and enhance the emission of Tb3+ significantly. PMID- 15818934 TI - [A fluorometric method for the determination of norepinephrine in tissue]. AB - This paper reported the determination of norepinephrine (NE) by spectrofluorometry in tissue. The linear relationship between the flecoroscence intensity and the content of NE was in the range of 10-100 ng (lambdaex/lambdaem = 410 nm/505 nm). The rate of recovery is 91.68%-96.64% and RSD (n = 10) is 6.1%. This method is simple, rapid and sensitive. PMID- 15818935 TI - [Study on the molar absorption coefficients of Bi(III)-thiocyanate complexes in UV spectrum]. AB - The absorption spectra of bismuth(II)-thiocyanate in the Tween-80-H2O solution were studied in this paper, in which the maximum wavelength lambdamax = 335 nm and the molar extinction coefficient epsilon335 = 2.61 x 10(4), similar to those of complexes of bismuth with PAR or azoarsenic(III), epsilon610 = 2.70 x 10(4). A new tribasic complex was found and used to explain that bismuth(III)-thiocyanate is a sensitive colour reaction in the Tween-80-H2O solution. PMID- 15818936 TI - [Study on the thermodynamical molar absorptivity of the interaction of cobalt(II) and the histidine]. AB - In this paper UV-Vis absorption spectrum of Cobalt(II) with the histidine and the thermodynamical molar absorptivity of the complex reaction were determined and the complexes of Cobalt(II) with the histidine were compared in terms of stability. PMID- 15818937 TI - [Study on the action force of dimerization of cobalt tetrasulfonate Ph thalocyanine in aqueous solotion]. AB - The dimerization of cobalt tetrasulfonate phthalocyanine (CoTSPc) was studied in the microemulsion, micelles and aqueous alcoholic solutions. The coordination reaction for the formation of complexes CoTSPc (L), which was formed by cobalt tetrasulfonate phthalocyanine (CoTSPc) plus ligand L(L = NH3, CN-, en), was also studied. The reaction kinetics was discussed and the stability constant K of CoTSPc(L)2 was also calculated from absorbance measurement. PMID- 15818938 TI - [Association and isochromatization of ion-pair of the acid and basic]. AB - In this paper, the method for forming the isochromatic dye ion-pair was described, and the mechanism and the types of the association of the ion-pair studied. The isochromatization of the associated reaction and the solvent effect were discussed by the electron theory with the absorption spectrum and energy diagram. The results of the extraction spectrophotometric method of the isochromatic dye ion-pairs were shown in a table, indicating the high sensitivity of the isochromatic dye ion-pairs. It is proven that the method of the extraction spectrophotometry, the flotation spectrophotometry and the extraction fluorometry was of high and superhigh sensitivity. PMID- 15818939 TI - [A new method for the determination of captopril]. AB - Captopril was determined in medicine by a new photometric method based on the color formation of Cu(I)-neocuproine complex from neocuproine and Cu(II) which could be reduced by captopril. The method was simple and rapid, and the results obtained were in agreement with those of tungsten blue method. PMID- 15818940 TI - [A multichannel laser Raman spectral detecting system]. AB - A multichannel laser Raman spectral detecting system is described. It consists of Ar+ laser, outer optics, polychrometer and optical multichannel analyzer (OMA II). Raman spectra of CCl4, C6H6 and C6-H12 were detected. Tests showed that the time required to acquire a Raman spectrum with this/multichannel system is greatly reduced, but the integral time per channel can be expanded to 10-50 s in contrast with a conventional single channel Raman spectrometer. When the working temperature of the multichannel detector is dropped to 5 degrees C, the Raman spectra with high signal to noise ratio, high sensitivity and good quality can be obtained. Processing Raman spectral curves with differential software, the Raman lines which are nearly submerged in Rayleigh line can be picked up. PMID- 15818941 TI - [Detection of the optimum resolution of monochromator]. AB - The relationship between resolution limit and step-motor parameter in monochromator has been discussed by optical method, which offers an experimental method for optimum utilization of the resolution limit and some reference example for improving resolution by using a step-motor. PMID- 15818942 TI - [The 1. 54 microm laser and upconversion luminescence of YELPP noncrystalline induced by 969 nm LD laser]. AB - In this paper, the absorption of Er3+, Yb3+ penta-phosphate noncrystalline (YELPP) was measured and the basic spectral parameters were calculated. Adopting the longitudinal-pump method with a domestic diode laser (LD), we achieved CW 1.5 microm chipped laser in China for the first time. The power of 1.5 microm laser was quite stable. The target of 1.5 microm laser reached the international level of early 1990s' in this field. In addition, the up-conversion luminescence was measured under the condition that 1.5 microm laser was either oscillating or not. The relationship between 1.5 microm chipped laser and up-conversion luminescence was analyzed initially. PMID- 15818943 TI - [The energy levels and oscillator strengths for transitions 3d(10)4s-3d94s4p in ions from Cu I -As V]. AB - The energy levels of configurations 3d(10)4s and 3d(9)4s4p of copper-like ions from Cu I to As V were calculated by means of the configuration interaction theory and parametric fit interpolation method. Calculated energy levels of 3d(9)4s4p are all in good agreement with experimental values . Most energy levels of ions Zn II and Ga III are sheer predicated ones. Also, the transition wavelengths and corresponding oscillator strengths of 3d(10)4s-3d(9)4s4p were calculated. PMID- 15818944 TI - [Study on laser flash spectroscopy of laser-induced interaction of DNA and polynucleotides with SO4*-]. AB - The interactions of DNA and polynucleotides (poly[G],poly[A] and poly[C]) with SO4*- were investigated by 248 nm laser flash photolysis. The results show that, the reactions of poly[G] or poly[A] with SO4*- yield predominantly the corresponding one-electron-oxidated base radicals. With poly[C], it is suggested that SO4*- radical reacts predominantly by addition to the base moiety to produce C5-yl and C6-yl sulphate radical adducts,and subsequently H-abstraction yields C2'-sugar radical. The predominant species resulting from the reaction of DNA with SO4*- is DNA guanyl radical. PMID- 15818945 TI - [Double resonance multiphoton spectrum of high excited state of iodine]. AB - The F0u(+) ion-pair state of I2 was studied in detail by using optical-optical double resonance multiphoton ionization spectroscopy technique. The OODR-MPI spectra were analyzed and identified based on the molecular constants published. The progression with a spacing of 37 cm(-1) at T(e) = 52 400 cm(-1) is identified to be the highly vibrational levels of F0u(+) state, and the transition pathway of excitation and ionization by multiphoton was determined. PMID- 15818946 TI - [The synthesis and absorption spectrum of [Fe(C60)] x ClO]. AB - The complexes of [Fe(C60)] x C10 and [Fe(C70)] x ClO containing C60 (C70) were obtained by mixing and grinding solid C60 (C70 contained) and FeCl3 x 6H2O. The complexes are dark brown, and dissolve in water. PMID- 15818947 TI - [Progress of surface enhanced Raman spectroscopic studies in nonaqueous systems]. AB - In this report, we describe the applications and theoretical aspects of SERS phenomenon in nonaqueous systems, which mainly involves the structure of solid/nonaqueous system, the adsorption of some neutral and organic molecules, inorganic ions and other complex compound molecules, the process and mechanism of some electrochemical oxidation reduction reactions in organic solvents etc.. In this review, we also depict the similarities and differences in SER spectra of aqueous and non-aqueous systems. PMID- 15818948 TI - [The investigation for lysozyme conformation changes in deuteromethanol solution by FT-Raman spectrometer]. AB - Lysozyme conformation in deuteromethanol-deuteroxide solution has been studied by FT-Raman spectrometer. By using amide I mode in the Raman spectra and curve fitting method, information of Lysozyme's secondary structure is easily detected in details, and tertiary structure changes can be suggested by relative intensity changes of some bands. It is concluded that lysozyme conformation may be changed to loose structure with incrasing deuteromethanol concentration, and it seems like that increasing loose beta-sheet content lead to the gel's appearing in the solution. PMID- 15818949 TI - [Surface enhanced Raman scattering of thiourea in acidic solution]. AB - Surface enhanced Raman scattering of thiourea adsorbed on the plated silver surface in an acidic solution has been studied. The results show that the thiourea molecules are inclined to adsorbed on the silver surface via sulphur. When the concentration of HCl in the solution is increased, the plane of the thiourea molecule deviates more from the normal line of the silver surface. The experimental results also suggest that the rather strong chemical enhanced mechanism would exist in this system. PMID- 15818950 TI - [Direct determination of raw plant drugs by FTIR]. AB - Twenty-five common raw plant drugs were determined quickly and directly with diffuse reflectance drawing sample of FTIR for the first time. The results show that the drugs with different chemical compounds are characteristics of different peaks. The category of plant drugs may be firmly identified through their respective IR characteristic absorption spectrum. Furthermore, the approach can be achieved quickly and accurately without extraction and separation of samples. PMID- 15818951 TI - [A Raman spectral studies of rare-earth (REE) fluoro-carbonate minerals]. AB - The Raman spectra of REE fluoro-carbonate minerals were studies by a type of RIT 30 laser Raman spectrometer. On the basis of Raman spectral properties, REE fluoro-carbonate minerals can be divided into three groups: Ba-REE fluoro carbonate mineral, Ca-REE fluoro-carbonate mineral and REE fluoro-carbonate mineral. Obviously, a differential kind of mineral is different in the vibration frequency of the Raman spectra. They are as follows: Cordylite v1-1 088, v2-967, v3-1 538, v4-720, 628; Hauheite v1-1 089,v3-1 525, 1 596, v4-720, 649; Cerbaite v1-1 088, v2-911, v3-1 516, v4-718,625; Parisite v1-1 095,1 075, v2-915, v3-1 461, v4-744, 732; Bastnesite v1-1 098, v2-835, v3-1 476, 1 447, v4-732. PMID- 15818952 TI - [Spectral properties of broad emission bands of Cr3+ in A3M2Ge3O12 (A = Cd2+, Ca2+; M = AI3+, Ga3+, Sc3+) garnets]. AB - The spectral properties of the Cr3+ broad emission band in A3M2Ge3O2 : Cr(A = Cd2+ ,Ca3+; M = Al3+,Ga3+,Sc3+) germanate garnets at room temperature are reported for the first time. In the order of Al3+ --> Ga3+ --> Sc3+ and Cd2+ --> C2z+ ions which occupy the octahedral and duodeahedral sites, respectively, R-NIR broad emission band of 4T2 --> 4A2 level transition, emission peak, and the spectral long wavelength and short wavelength sides of Cr3+ shift to long wavelengths having lower energy. The reason is that the crystal field strengths in the octahedral decrease, while the cation ionic radii in the A3M2Ge3O12 system garnets increase gradually. The R-NIR emission intensity of Cr3+ is increased with some Sc3+ ions substitution for Al3+ or Ga3+ ions in octahedral sites. PMID- 15818953 TI - [The spectral study of the surface modified medical rubber]. AB - In this article ,the drug-resistance of two kinds of medical rubber whose surfaces have been modified were investigated by ATR-FTIR and XPS. The experimental results show that the compositions of the two samples'surface and body are different. The surface is fluorinated rubber although the body is butyl rubber. The ratio of fluorine to carbon atom in sample Ii -1 is higher than that in sample I -1. The principal join between F and C is the form--CF2--in sample II -1,but in sample I -1 it is the form--CF2-- and--CHF--. The change for F/C of the different depth in sample II- 1 was relatively less than that in sample I -1 when they were etched by argon ion bundle in the same conditions. PMID- 15818955 TI - [Detection of low temperature rheologic behavior of lubricating oil by FTIR]. AB - The composition and structure of lubricating oil expressed by FTIR were used for its characterization. Artificial nervous network was used as a mathematical model. CCS-15C viscosity of base oil was forecasted and good result was obtained. The result has shown that the artificial nervous network is an effective mathematical model for studying the relation between the composition and structure of lubricating oil and its performance. PMID- 15818954 TI - [Effects of anion ions on the structures of lanthanum glycine amino acid complex compounds]. AB - Two lanthanum-glycine amino acid complexes, La(Gly)3 (ClO4)3 x H2O and La(Gly)3Cl3 x 3H2O, were synthesized with different anion ions. The infrared spectra of two complexes were characterized. The results show that the two complexes have different structures with different anion ions and the water molecules exit with different status in the two complexes. The results also indicate that one of the perchlorate ions is involved in the coordination in La(Gly)a(ClO4)3 x H2O. PMID- 15818956 TI - [Studies on the infrared and fluorescence spectra of europium lanthanum-benzoate complexes]. AB - A series of mixed rare earth complexes of benzoic acid with europium and lanthanum were synthesized by the reaction of trivalent rare earth chlorides with ammonium benzoate in aqueous solution. Their composition is expressed with the general formula (Eu(1-chi)La(chi)) L3, where L = C6H5COO(-1), chi = 0.0-1.0. Their infrared and fluorescence spectra were investigated. The characteristic asymmetric (v(as)) and symmetric (v(s)) stretching vibrations of carboxyl group for rare earth complexes shift to lower frequency in comparison with sodium benzoate, and both the absorption bands show splitting. In addition, the difference between v(as) and v(s), deltav (120-125) cm(-1), is lees than that of sodium benzoate (deltav =137 cm(-1)). These situations show that the coordinating pattern of the carboxyl groups in the complexes is the form of chelation bridging, therefore, it may be suggested that the rare earth (Eu, La) benzoates are a heteronuclear polymers. The dependence of the relative fluorescence intensity of the mixed rare earth complexes upon the content for La3+ ions was discussed. The results show that the emission intensity for Eu3+ ions can be greatly enhanced if a part of Eu3+ ions in europium benzoate were substituted by La3+ inos. When chi = 0.4, 0.5 or 0.7, the fluorescence intensity of the rare earth complexes can rise above 40%. PMID- 15818957 TI - [Optical spectral verification of the phthalanilic acid]. AB - The structure of an unknown compound was studied by NMR spectrum and IR spectrum. The compound was proved to be phthalanilic acid. The result provids useful information for studying biological activity and biological course of this matter. PMID- 15818958 TI - [Fluorimetric study of Tb-EDTA-quercetin and determination of quercetin]. AB - Tb-EDTA-Quercetin complexes were formed under basic conditions. Intramolecular energy transfer occurs in the Tb-EDTA complex with quercetin (QCT) and the terbium (III) ion fluorescence can be sensitized by QCT. Accordingly a fluorimetric method was used for the determination of QCT in urine, serum and mixed substance of rutin-quercetin. The method is simple, sensitive and fast. The linear range is 5.0 x (-7)-1.0 x10(-5) mol x L(-1) and the detection limit is 5.0 x 10(-8) mol x L(-1). This method can be applied to the determination of QCT in non-pretreated urine and serum successfully. The recoveries are in the range of 103%-96%. PMID- 15818959 TI - [Study on the ternary complex of CPFX, copper (II) and DNA by the method of fluorescence]. AB - Using the method of fluorescence spectrometry, the formation of binary complexes of CPFX-Cu(II) and CPFX-eucaryote DNA, and ternary complexes of CPFX-Cu(II) eucaryote DNA was studied. The results show that the fluorescent intensity of CPFX can be quenched in the presence of Cu(II ) or DNA in CPFX system. In the presence of Cu (II), the fuorescence quenching action of DNA on CPFX was enhanced strongly. Based on the fluorescent intensity, the formation constants of CPFX-Cu (II) and CPFX-DNA in either the presence or absence were calculated,and the mechanism of interaction among CPFX,Cu(II) and DNA was presented. PMID- 15818960 TI - [Spectroscopic study on procaine hydrochloride and its cyclodextrins inclusion complexes]. AB - In this paper, by means of the ultraviolet spectrum of procaine, we showed the existence of two types of procaine (molecular and ion types) and determined their pH values. The variations of fluorescence intensity of the two types in the presence or absence beta-CD, HP-beta-CD and Methyl-beta-CD were studied. PMID- 15818961 TI - [Fluorescence quenching method for the determination of trace zirconium with 5' nitor-salicylfluorone and hexadecyltrimethylammonium bromide]. AB - A new fluorometric method has been developed for the determination of trace zirconium, based on the fluorescence quenching effect of Zr (IV)-5'-nitro salicylfluorone (5'-N-SAF)-hexadecyltrimethylammonium bromide (CTMAB). In the H2SO4 medium and in the presence of CTMAB Zr (IV) reacts on 5'-N-SAF to form a 14 orange complex. Its maximum excitation wavelengths are 475 and 505 nm,and the maximum emission wavelength is 525 nm. The linear range is 2.0-80 microg/L, and the detection limit is 2.0 microg/L. The method has been applied to the direct determination of trace Zr (IV) in the aluminium alloy reference sample with satisfactory results. PMID- 15818962 TI - [A new absorbance ratio-derivative method and its application to simultaneous determination of two components in a mixture]. AB - In this paper, the principle of a new absorbance ratio-derivative method was described. The experimental methods of derivation with subtraction technique on ordinary spectrophotometer and simultaneous determination of phenol and resorcin were studied. The result is satisfying. The recovery of resorcin and phenol is in the range of 99.4%-104.2% (n = 25, RSD = 1.1%) and 97.1%-102.8% (n = 25, RSD = 1.6%) respectively. PMID- 15818963 TI - [Determination of GeO2 in germanium-132 by phenylfuorone]. AB - Inorganic (GeO2) in carboxyethyl germanium sesquioxide (germanium-132) was determined by phenylfuorone spectrometry. The method is convenient and has good precision and accuracy. The recovery of GeO2 is 93%-107%. PMID- 15818964 TI - A new generation of instrumentation and capabilities for atomic mass spectrometry. PMID- 15818965 TI - [Information system for ICP-AES in windows environment]. AB - An information system for inductively coupled plasma atomic emission spectrometry (ICP-AES) in MS Windows environment was developed based on the previous work in the labroatory. The system contains the data of about 28 000 spectral lines and a function of ICP spectral simulation, so it would be very helpful for line selection. The system also contains the Kalman filter and factor analysis programmes written with MS Visual Basic(version 4.0), which can be used for spectral interference correction and peak position optimization. A large amount of real spectral scanning data of rare earth elements were included in the system for user's references. All these characteristics made the system more useful and practical. PMID- 15818966 TI - [Determination of lead in tinplate by ICP-AES]. AB - The determination of lead in tinplate by ICP-AES was presented in this paper. The tinplate samples were pretreated with NaOH-H2O2 for dissolving tin and lead,and the spectral interferences from iron could thus be avoided. The lead content in the sample was determined in the range of 0.14-0.38 microg/cm2. The recovery and the relative standard deviation (RSD) were in the ranges of 96.1%-103.2% and 2.6% 3.8%, respectively. The method was simple,rapid and efficient. PMID- 15818967 TI - [Determination of 14 elements in chromium-rich brewer's yeast by ICP-AES]. AB - An ICP-AES method was established for the determination of 14 elements in chromium-rich and normal brewer's yeast. The RSDs of two standard reference materials were 0. 5%-3.2% and 0.4%-3.4%, respectively. The results showed that chromium contents could affect the concentrations of other elements in chromium rich brewer's yeast. The concentrations of K, Mn, P, V, Mg and Ca in chromium rich brewer's yeast were significantly lower than those in normal brewer's yeast. PMID- 15818968 TI - [Study on the elements in cosmetics by microwave digestion-iCP-AES I. The determination of Pb, As, Cr, Cd, Sr, Bi and Se in solid state cosmetics]. AB - A safe and effective method for the microwave digestion of solid state cosmetic samples is described. The overall digestion conditions, including pressure, power, acids and time of digestion were evaluated by analysing Pb, As, Cr, Cd, Sr, Bi and Se of routine samples with ICP-AES. This method was applied to the determination of certified materials with good agreement. PMID- 15818969 TI - [Microwave acid-digestion ICP-MS method for the determination of sixteen elements in marine products]. AB - Microwave acid-digestion samples and ICP-MS method were established for the determination of Na, Mg, Al, P, Cr, Mn, Fe, Ni, Cu, Zn, As, Sr, Mo, Cd, Pb and Hg in marine product. According standard sample GBW08571 mussel, we cheek the method accuracy and precision. The accuracy average values are 96.7%. These results are very suitable for the reference values. The RSDs of all elements are less than 5%. We use several internal standard correction method to correct the matrix interference. The method is simple, rapid, accurate and reliable. PMID- 15818970 TI - [Method for the determination of trace Li in serum of cancer patients by tungstate-coated graphite furnace atomic absorption spectrometry]. AB - Using a tungstate coated graphite tube. The sensitivity of Li was significantly increased and trace Li in serum samples was determined by Zeeman graphite AAS with (NH4)2SO4 as a matrix modifier. The effects of different metal ions and biological matrix were eliminated. The characteristic concentration is 5.53 x 10( 3) microg x mL(-/1)%. The recovery is greater than 93.72%. The relative standard deviation is less than 8.93%. PMID- 15818971 TI - [Determination of beryllium in river water by graphite furnace atomic absorption spectrometry]. AB - The graphite tube such as Lanthanum-coated or Barium-coated have been tested for the determination of Be in water by GFAAS. The effects of different metal ions have been eliminated by adding Al (III) or Ca (III) as matrix modifier. The method is handy and fast. The recovery is 92.6%-105.0% and RSD is less than 10.24%. PMID- 15818972 TI - [Determination of trace selenium in spirulina by continuous-flow hydride generation atomic absorption spectrometry]. AB - The mixed solutions of the hydrogenized element-NaOH-NaBH4 and HCI solution were fed into hydride vapor generator at the same rate by multi-flow peristaltic pump. Selenium element in the aqueous solution reacted on the nascent hydrogen to generate gaseous hydride. No separation was needed and the method was highly sensitive with little interference. The selenium content in spirulina was determined and a reliable analytical method was established. The determination limit of the method was 0.017 microg x g(-1), and the recovery of the samples was 98.89%-101.20%. PMID- 15818973 TI - [Measurement of cadmium content in soils and vegetables by FAAS]. AB - The analytical and testing centre the total cadmium (Cd) and the available Cd contents in different Cd-polluted soils were measured by FAAS. The Cd content in the stems and leaves of cole was tested in the same way. The methods of sample pretreatment for the total Cd and available Cd in soils and in cole were compared respectively. The standard sample recovery rates were all above 96 percent, and the relative standard deviations were less than 3.97 percent. The FAAS is rapid and accurate. PMID- 15818974 TI - [A Zeeman graphite furnace atomic absorption spectrometric method for the determination of trace copper and chromium in drinking water]. AB - The determination of trace copper and chromium in drinking water is described in this paper using transverse heated graphite atomizer (THGA) with the technique of Zeeman effect background correction without any other matrix modifiers. The method is fast, and simple with low detection limit which makes it possible to be used for routine analysis of drinking water. PMID- 15818975 TI - [Study on the determination of trace caesium in bittern by extraction separation graphite furnace atomic absorption spectrometry]. AB - The atomizatiom behavior of Cs in bittern and the mechanism were studied by graphite furnace atomic absorption spectrometry. Interferences of inorganic acid from the bittern on absorption signal of Cs were examined. It is shown that there are gaseous interferences and molecular absorption interferences and ionization interferences with regard to the interference of chlorides on absorption signal of Cs. The effects of 3 crown ethers, 5 big-anion reagents and chlorides on extractability Cs have been compared. A extraction separation GFAAS method for determining Cs in bittern has been established by pyrolytic graphite tube, KNO3 NH4NO3 matrix modifier and 18-C-6, bromothymol blue, chloroform extraction separation. The method exhibits high sensitivity and selectivity, it has been used for the determination of trace Cs in bittern. The characteristic mass is 1.4 x 10(-10) g/0.004 4 A. The adding recovery is 89%-105% and the relative standard deviation is 4.13% (n = 13). PMID- 15818976 TI - [Study on pretreatment for the determination of several nutrition element in eggs with AAS]. AB - In this paper, 0.1 mol x L(-1) HCl, 6 mol x L(-1) HCl, pH 7.0 1 mol x L(-1) NH4Ac was oscillated and 6 mol x L-' HCI was boilled to extract and determine K, Na, Ca, Zn, Fe, Mn, Mg and Cu. Comparing with classical method that popularly used HNO3-HCl04 to degest and ash samples,the results indicated that for macroelement K,Na and Mg several extraction methods are similar to classical method,the recovery was 90%-115%,for microelement Zn, Mn and Fe could be extracted using pH7.0 1 mol x L(-1) NH4Ac, 0.1 mol x L(-1) HCI, 6 mol x L(1) HCI oscillating,the recovery was 100%-115%. Cu could be analysed using 6 mol x L(-1) HCI boiling. But the results of Ca were not better because of the enhancement effect. The results with ash sample were lower because many elements were lost when ash temperature was up to 450 degrees C. These methods of exatraction pretreatment are simple and rapid. The recovery and precision are satisfactory. PMID- 15818977 TI - [Determination of K, Na and Ca in alizarin by flame atomic absorption spectrophotometry]. AB - This paper reports the development of the determination of trace metals K, Na and Ca in alizarin. The method is convenient to determine elements in alizarin by flame atom absorption spectrophotometry. The results show that the method has good precision and accuracy. The relative standard deviations of K, Na and Ca are 4.4%, 2.5% and 3.7%, respectively, and the rates of recovery are 97.5%, 98.3% and 98.1% respectively. PMID- 15818978 TI - [Continuous determination of Ca and Mg in tea by flame atmic absorption spectrophotometry]. AB - In this paper the Ca and Mg in tea were detected using continuous determination by FAAS. The method is simple, rapid and has good precision and accuracy. The relative standard deviation is 1.87%-2.75%. The recovery rate is 92.5%-105.1%. PMID- 15818979 TI - [Research into simultaneous determination of five components by ridge regression spectrophotometry]. AB - Ridge regression spectrophotometry (LHG) is one of the computational spectrophotometry developed recently. It is used in this paper to the analysis of five component s--acetaminophen, p -aminophenol. p -nitrophenol. p acetaminophenol acetic acid ester and p -nitrophenol acetic acid ester. The choose of medium and the determination of the amount of mixed standard solution were described in detail. The experimental results show that in the medium of 0.1 mol x L(-1) HCI, the recovery ratio of each component ranges from 95.5% to 106.1% and the RSD ranges from 0.8% to 9.8%. This approach is simple and suitable for the computer-aid analysis. The approach has been used to the analysis of the sample in the course of synthetic acetaminophen. PMID- 15818980 TI - [Determination of 2,4-dinitrochlorobenzene in environmental water by primary secondary wavelengths new spectrophotometry]. AB - In basic solution, dinitrochlorobenzene may react on pyridine to form a purplish red compound. In this paper with the absorption property of a suspended liquid against light a new analytical method as primary-secondary wavelengths spectrophotometry was studied to determine trace dinitrochlorobenzene in environmental water. In 0-50 microg dinitrochlorobenzene/25 mL, the calculation curve is very stable and hardly affected by environment conditions. Experiments for practical waters have shown that the detection limit was 0.7 microg/25 mL dinitrochlorobenzene and RSD < or = 2.7% and the recovery rate was 99.2%-101.0%. PMID- 15818981 TI - [Study on catalytic spectrophotometic determination of nitrite with catechol violet system]. AB - A new sensitive method for the determination of trace nitrite was developed, based on nitrite as a catalyst. The optimum kinetic conditions were discussed and the concentration of nitrite was linear in the rang of 0.2-3.5 microg/10mL. The proposed method is simple, rapid, selective and convenient to determine nitrite in water and vegetables with good results. PMID- 15818982 TI - [Determination of analginum in medicine by ironic rhodanate spectrophotometry]. AB - Analginum in preparations was determined by the ironic rhodanate fading spectrophotometric method, based on Fe3+ reduced into Fe2+ by analginum and reacting on potassium rhodanate. The obtained results were in agreement with those of the iodine titration method and the ultraviolet photometric method. PMID- 15818983 TI - [The two-step three-photon resonant ionization of atom Cu]. AB - This paper discusses the process of resonant laser ablation (RLA), where the two step three-photon resonant ionization of copper is considered as the best scheme. The rate equations of resonant ionization of copper are calculated and the factors that affect the ionization efficiency are discussed. The results prove the potential advantages of RLA especially the low energy threshold. PMID- 15818984 TI - [Recursion formula of energy levels for 2p(3)3s and 2p(3)3d of O I isoelectronic series]. AB - Based on the perturbation expansion and the observed energy levels for 2p(3)3s and 2p(3)3d of O I -like ions, a recursion formula for the energy levels has been derived. It is found that this method is considerably simple and accurate. PMID- 15818985 TI - [Pattern recognition applied to the Pu I even-parity energy level]. AB - The partial least square (PLS)-back propagation network (BPN) method is applied to assigning the electronic configurations of Pu I even-parity levels and the assignments agree well with the experimental results. It is shown that PLS-BPN can be used as a tool to assign complicated spectral levels. PMID- 15818986 TI - [Studies on data compression of 1-D NMR spectra using wavelet transform]. AB - The multiresolution signal decomposition (MRSD) algorithm based on wavelet transform was applied to the compression of 1-D NMR spectra. A NMR spectrum which was composed of 32768 data points was investigated and the spectrum can be completely reconstructed with the residual square deviation (RSD) being as small as 2.7769 x 10(-4) when the spectrum was compressed to 512 data points. The relationship among the RSD and the different wavelet function, number of decomposiion and the compression ratio was also discussed. PMID- 15818987 TI - [X-ray photoelectron spectroscopy and mass spectrum of organic tin compounds]. AB - The relationship between the chemical displacement of the binding energy and the different chemical environment for 12 organic tin compounds was studied by means of X-ray photoelectron spectronscopy. The different substituents in the compounds have influence on the tin outer electron and Sn-O bond, which was discussed by X ray photoelectron spectroscopy and mass spectrum. PMID- 15818988 TI - [Stimulated radiation from high-lying state of Li2 generated with multiphoton in a wide wavelength region]. AB - The experimental results of ultraviolet diffuse-band of stimulated radiation from the high-lying state of Li2 generated with any pumping wavelength between 620.0 665.0 nm were reported in this paper. In Li2-Li system, the high-lying state of Li2 was generated by two-photon excitation, ionization recombination process and collision energy pooling process, and the ultraviolet diffuse-band radiation was stimulated. The radiation was due to the transition from one of the comparable high-lying treble states to the low-lying repelling state a 3sigma(+)u. The excitation mechanisms involved were discussed in detail. PMID- 15818989 TI - [The Raman spectra of monohydrate citric acid]. AB - The Raman Spectra of monohydratecitric acid in different geometric arrangements have been reported in this paper. From the shift of some spectra, the anisotropic Raman scattering and depoearization effects, we can conclude that there exist hydrogen bonds in the crystal. Because of the effect of hydrogen bonds, electrical coupling between molecules causes the C-C, C-O bonds to possess some double bond character. On the other hand, the C=O bond possesses some single bond character. PMID- 15818990 TI - [Studies on tailed metalloporphyrin coordination compounds. IX. Synthesis of tailed porphyrin with covalently linked phenylalanine and its fluorescence property]. AB - A new tailed porphyrin with covalently linked phenylalanine 2 and its zinc complex 3 were synthesized. Intramolecular energy transfer (from singlet excited phenylalanine to porphyrin) was observed in compound 2 and 3. The influence of coordination chemistry factor on the fluorescence of porphyrin was also discussed. PMID- 15818991 TI - [The investigation of photoacoustic spectroscopy for polydiacetylene Langmuir Blodgett films. (I) Structure characterization and photoacoustic absorption coefficient spectra]. AB - The polydiacetylene (PAD) Langmuir-Blodgett (LB) films, which have potential applications in industry, were studied by using X-ray low angle diffraction and photoacoustic chopping frequency effects and photoacoustic absorption coefficients obtained. The X-ray low angle diffraction of PDA LB films prepared by different sequence of polymerization/transferring shows that the two kinds of PDA LB films have different array structures. The chopping frequency effects of PDA LB films of different thickness are consistent with Roscenwaig-Gersho theory and show that the thermal property of PDA LB films gradually becomes obvious with the increase of thickness. A new method of acquiring photoacoustic absorption coefficient from photoacoustic spectroscopy was also developed. The achievement of this paper shows that the PAS technique is a valid means for studying LB films. PMID- 15818992 TI - [Artificial neural network application in spectral recognition]. AB - An artificial neural network was devised and established for spectral matching. It was proved that when training sample spectra and unknown spectra had the same magnitude calibration, the network recognized spectra very well and had some advantages over other conventional spectral matching methods. PMID- 15818993 TI - [FTIR and FT-Raman studies on interactions between water and surfactants in water/KDEHP (75%)-HDEHP (25%)/n-HEPTANE microemulsion systems]. AB - The conformation of the hydrocarbon chains in water/KDEHP(75%)-HDEHP(25%)/n HEPTANE microemulsion systems was studied by FT-Raman spectroscopy. The increase in the intensity ratio Ir (I2937/I2875) showed trans to gauche transformation. The hydration of the polar heads in microemulsions was investigated by FT-IR spectroscopy. Some probable structures of KDEHP and HDEHP in microemulsions were proposed. PMID- 15818994 TI - [SERS of bilirubin and its complexes]. AB - SERS spectra of bilirubin and its complexes were determined in Ag sol by the technique of the connection between NIR-FT-Raman and SERS. The spectra showed that this kind of biological complexes, which is closely related to gallstones, differ in coordination pattens and adsorptive orientation as well. The reason why the black-pigment gallstone exhibits that peculiar dark color was also given from the coordination-chemistry point of view. PMID- 15818995 TI - [Water structure in water/KDEHP-HDEHP/n-Heptane microemulsion by FTIR spectroscopy]. AB - The structure of water in microemulsion is one of the basic subjects in the field of surface science. FTIR spectroscopy is considered as a powerful tool in the study on water structure. In this paper, we investigated water structure in the water/KDEHP (potassium bis (2-ethylhexyl) phosphate) -HDEHP/n-Heptane system. By using the Fourier self-deconvolution and curve-fitting method, the OH stretching band of water in the KDEHP-HDEHP microemulsion was resolved into four subpeaks which were tentatively assigned to absorbances of four different components of water. In order to explain the variations of peak positions of subpeaks with water content some probable hydration models of surfactants were proposed. PMID- 15818996 TI - [Study on the Co(DC)2-NaDC mixed micelle system]. AB - Eight coordinated complex samples of CoCl2-NaDC were obtained,using various concentration and initial molar ratio of CoCl2 and NaDC. They exhibited distinct appearances and characters. Their different structures and components were investigated by FTIR spectroscopy, X-ray diffraction analysis and ICP analysis. We can give the following conclusions: (1) when the concentration of NaDC is smaller than C.M.C, Co2+ ions react with mono-molecular NaDC and the component of the pink wadding-like resultant is Co(DC)2 x 3H2O. (2) When the concentration of NaDC is larger than C.M.C, and the concentration of CoCl2 is higher (0.5 mol/L), Co2+ ions destroy NaDC micelles, and the component of the pink wadding-like resultant is also Co(DC)2 x 3H2O. (3) When the concentration of NaDC is larger than C.M.C, and the concentration of CoCl2 is lower (0.1 mol/L or more lower), Co(DC)2 and NaDC may form mixed micelles, and some of NaDC micelles connect with each other through the Co2+ bridges to produce complicate Co(DC)2-NaDC complex with gel character and macromolecular network structure. In human body,the concentration of metal ions is usually lower and the concentration of NaDC is larger than C.M.C, So the third kind is suggested as an ideal model of the interaction between Co2+ ions and NaDC in vivo. PMID- 15818997 TI - [The absorption spectrum of C70]. AB - With the methyl benzene as solution,ultraviolet and visible absorption spectra of the mixture of C60 and C70's methyl benzene saturant solution as specimen, infrared absorption spectrum of the mixture of C60 and C70 was measured. The absorption spectrum of C70 was determined by comparison. PMID- 15818998 TI - [The formation and oxidization of C60/C70 in graphite are discharge process]. AB - It was found by means of infrared absorption spectrum that in the course of producing C60/C70 by graphite is discharge, a large quantity of C60/C70's oxide was produced, and such oxide features semiconduction and has advantage of good thermostability in vacuum. PMID- 15818999 TI - [Investigation of the reaction of iso-propylamine on propylene oxide by Fourier transform infrared spectroscopy]. AB - Fourier transform infrared spectroscopy method has been developed for in-situ monitoring the reaction of iso-propylamine on propylene oxide without the need to sample the reaction medium. The profile of the reaction and the influences of conditions can be directly reflected in the feature absorbance changes of the reactant and product at 826 cm(-1) and 1069 cm(-1), respectively. It is shown that the in situ infrared spectroscopy is a powerful technique to investigate the mechanism and kinetics of the reaction and to optimize the control conditions. PMID- 15819000 TI - [Study on spectral emission characteristics of infrared lamps]. AB - The spectral characteristics at wavelength ranges of 0.35-0.85 microm and 1.72 16.7 microm (5800-600 cm(-1)) for both domestic and imported infrared lamps used in scientific research, industry, medical service, home electrical appliance etc. have heen studied and compared. This paper has provided the theoreical and experimental bases for their applications and improvement in designs, technology of manufacture, quality and performance. PMID- 15819001 TI - [The study of castor oil sulphate synthesis process by infrared spectroscopy]. AB - The castor oil sulphate synthesis process was studied by infrared spectroscopy. Based on the determination of iodone value the reaction mechanism was discussed. Both the duplicated bond and the hydroxide ion in castor oil took part in sulphate reaction and a hydrolysis also took place in the castor acid. PMID- 15819002 TI - [Quick determination of lubricating oil additive by infrared spectrum method]. AB - Some additives were measured by PE-FTIR 1725 X instrument and "Quant" Software (PE company), with the relative error less than 10%. Several oil samples such as 15W/40SE, 15W/30 SG/CD, 15W/40CD and 15W/30CC were analysed in which the additive and base oil ranged from 0.2%-90%. PMID- 15819003 TI - [Ultra trace determination of platinum, palladium, gold and rhodium in minerals]. AB - Our work studied the minerals dissolved in aqua regia. The trace Pt(II), Pd(II) Au (III) and Rh(I) complexes were loaded by the mixed carbon powder which carried diphenyl thiourea when aqua regia or hydrochloric acid media as well as stannous chloride were present. We added some carbon powder in ashen loader and determined the elements with atomic emission spectrometry. The determination sensitivity counted with 10 grams of samples is 1 x 10(-9). The RSD for 0.05 x 10(-6) standard Pt, Pd, Au and Rh solutions are 6.0%, 5.1%, 8.5% and 11.4%, respectively. The method is simple and operation is easy. The results satisfy our requirement. PMID- 15819004 TI - [Simultaneous determination of phosphorus in hair by ICP-AES]. AB - A useful ICP-AES method for the determination of microelements P, Zn, Cu, Fe, Ca, Mg, Pb, Mn and Al in hair is described. The JY38 spectrometer in the JY70PII ICP AES system,equipped with a 3600 g/mm holographic diffraction grating (1 m focal length) was used for the determination of phosphorus. The detection limit value (at 3 standard deviations of background) for spectral line 213.618 nm is 0.022 microg x mL(-1). PMID- 15819005 TI - [Study of selenite bio-transformation in nature water by hydride generation atomic fluorescence]. AB - This paper studied selenite bio-transformation in nature water by hydride generation-atomic fluorescence. Production was primarily element selenium, also with volatile selenium. The added C sources accelerated the transformation. PMID- 15819006 TI - [Mechanism of arsenic hydride generation and atomization in low temperature atomic fluorescence spectrometry]. AB - With a hollow cathode lamp as exciting source, the optimum working parameters for the determination of As were selected. By adopting two hydride-generators to generate hydride and H2 simultaneously, the mechanism of AsH3 generation was studied. Experimental results show that the atomization process of AsH3 might be carried out mainly by collision with hydrogen radical. PMID- 15819007 TI - [Study on the determination method of Mo in the internal organs of body by graphite furnace atomic absorption spectrophotometry]. AB - A new method for the determination of Mo in the heart by tantalum-lining graphite tube was developed. The tantalum-lining graphite tube can entirely eliminate effects of stable carbide formed. The sensitivity was enhanced to compare with the conventional graphite tube. At the same time, it also climinates memory effect completely and reduces the interferences of coexistent elements such as Fe, Na, K etc. The method is simple, accurate and rapid. PMID- 15819008 TI - [Determination of ten Chinese traditional medicines made in Jiangxi province by AAS]. AB - The quantities of 21 microelements were determined in 10 Chinese traditional medicines, using both flame and graphite oven atomic absorption spectrometry. We studied the efficacies of such Chinese medicaments. The result showed the efficacies related to the inorganic microelements. PMID- 15819009 TI - [Determination of trace Co in XF-210 phosphono-acid dirt preventing agent by flame atomic absorption spectrophotometry]. AB - This paper reports the development of the determination of trace metal Co in XF 210 phosphono-acid dirt preventing agent by flame atomic absorption spectrophotometry. The method is convenient and the results show good precision and accuracy. The relative standard deviation was 1.42% and the rate of recovery was 95.7%. PMID- 15819010 TI - [Coprecipitation for preconcentration and determination of trace cadmium in solid waste phosphogypsum by FAAS]. AB - CuS was used as a collector precipitate to preconcentrate and separate trace cadmium in phosphogypsum. The optimum coprecipitation conditions were studied, it was found that Cd could be quantitatively coprecipitated with CuS in sample solution at pH 5.8-7.0 and that 100 microg of Cu was enough to coprecipitate Cd in 50 mL of sample solution. No interferences of coexistent elements were found. A sample was detected for 6 times. The RSD was 7.97% and the recovery was between 88% and 110%. The method was rapid, simple and satisfactory for detecting Cd in phosphogypsum. PMID- 15819011 TI - [Determination of potassium, calcium, manganese, chromium and cadmium in 10 kinds of (tobacco) cigarettes by flame atomic absorption spectrophotometer]. AB - We used the flame atomic absorption spectrophometer to determine the contents of potassium,calcium,manganese, chromium and cadmium in 10 kinds of (tobacco) cigarettes. The results showed that there were different contents of five kinds of metals in different cigarettes, providing useful data for plant and dryness of tobacco as well as the reduction of harm of smoking to human body. PMID- 15819012 TI - [Study of a new kind of spectrometer coupled with flow injection and CCD diode array detection for multi-components simultaneous determination]. AB - A new kind of spectrophotometric instrument combined with a flow injection device, a prism spectrography, a CCD diode array detection system and a popular microcomputer was established for multi-components simultaneous determination. The measurements of lead and cadmuium with meso-tetra (4-trimethylammoniumphenyl) porphine were completed with the instrument. The detection limit was 0.014 microg/mL for Pb(II) and 0.015 microg/mL for Cd(II). The sampling rate was 60 samples/h. PMID- 15819013 TI - [FPMULTI--a software for multi-layer composition and thickness analysis and its applications]. AB - Main features of FPMULTI, a software for analyzing composition and thickness of multi-layer samples simultaneously using XRF, are briefly described in this paper. Based on fundamental parameter method, the software has the capability of determining multi-layer samples containing up to 10 layers and 25 elements. Calibration standards can be bulk or multi-layer of pure element or multi-element standards. Application examples for tin-plates, hot dip galvanized zinc-plates and 'fingerprint-resistant' plates were given. FPMULTI is used to predict the relationship between intensities of different X-ray lines and the thickness first and then to analyze those samples. When using only few calibration standards, the results from FPMULTI are much better than those from linear regression method, and this reflects the advantages of the fundamental parameter approach. PMID- 15819014 TI - [Calculation and computer program of modified proportional factor method in XRF]. AB - This paper describes the principles and computer program of modified proportional factor method in XRF. All calculations were performed by computer program assembled using the true BASIC language, It is suited to analyzing major and minor elements in alloys. PMID- 15819015 TI - [Principle and application of scattering power function method in X-ray fluorescence analysis. III. Continuous scattering radiation]. AB - The study in this paper shows that the relationship between mass attenuation coefficient and continuous scattering radiation at wavelength 0.081 nm is not an inverse proportion supposed as the scattering internal standard method but a power function in a wide range of matix composition. Experimental results are in good agreement with that of theoretical prediction. A new method is recommended for matrix absorption correction using the 1.22 power function of continuous scattering radiation at wavelength 0.081 nm and used for microelement Sr determination in geological samples as an application example. The accuracy of the results by this method is nearly 4 times better than that from the traditional scattering internal method in a wide range of matrix composition. PMID- 15819016 TI - [Study on the human serums by absorption spectra]. AB - The absorption spectra of normal human and cancer patient serums were measured, and those of human serum after adding some cancer cells were also measured. The results showed some difference in absorbance among the three kinds of serums. Some useful results were obtained. PMID- 15819017 TI - [Fluorescence spectra of lanthanide coordination compounds]. AB - In this paper, the recent progress in fluorescence spectra of lanthanide coordination compounds including photophysics and photochemistry is briefly reviewed and the application of lanthanide complexes as fluorescent probes to structural analysis of coordination compounds and biomolecular systems is summed up. PMID- 15819018 TI - [Fluorescence studies on amoxicillin]. AB - The effects of pH, various organic solvents such as acetone, methanol, ethanol, surface-active agent on amoxicillin have been studied in detail. Put in pH=8 buffer solution, amoxicillin has strong fluorescence emission, excitaion wavelength of 277 nm, and emission wavelength of 303 nm. Calibration curve of amoxicillin was linear between 2 x 10(-5) and 2 x 10(-4) mol/L, with a regression coefficient of 0.9996. CTAB can enhance its fluorescence intensity. The results of fluorescence spectra of amoxicillin added with metal ions (Mg2+, Ca2+, Al3+, Ga3+) at pH=12 showed that there are interactions between amoxicillin and metal ions. PMID- 15819019 TI - [Absorption characteristics of the isochromatic dye ion-pair formed by tetrabromofluorescein and buthylrhodamine B]. AB - Absorption properties of tetrabromofluorescein (TBF) and buthylrhodamine B (BRB) and the isochromatic dye ion-pair formed by TBF and BRB were studied. The experiment showed that the proportion of TBF: BRB was 1:2 at pH=7 with Ksp = 2.16 x 10(-18), and the extraction rate was 84.2%. This provides a basis for the application of TBF and BRB to extraction spectrophotometry. PMID- 15819020 TI - [Determination of diclofenac sodium and codeine phosphate in shuerfen tablets by CPA-matrix method]. AB - The contents of diclofenac sodium and codeine phosphate in shuerfen tablets may be determined simultaneously by CPA-matrix method without separation of its components. The program was edited by BASIC. The average recoveries and RSD were 99.85%, 0.46% for diclofenac sodium and 99.80%, 0.50% codeine phosphate, respectively. This method is simple and rapid, and the result is accurate. PMID- 15819021 TI - [Study on spectrophotometric determination of trace nitrite with brilliant cresol blue]. AB - A new spectrophotometric method for the determination of trace nitrite was studied. The method is based on the diazo-reaction of brilliant cresol blue with nitrite. The linear range for the determination is 0.5-12 microg NO2(-)/25 mL. The detection limit is 0.5 microg NO2(-)/25 mL. The method has been used for the determination of trace nitrite in water. The mechanism of the reaction was discussed. PMID- 15819022 TI - [Spectrophotometric determination of iron using eriochrome cyanine R as extractant with phase-separation]. AB - Spectrophotometric determination of iron by using eriochrome cyanine R as extractant and color-developing agent in organic solvents was studied. The complex was extracted by PEG phase from the water solutions of pH4.5-6.5 (NaAc HAc). The maximum absorption of the complex is at 560 nm and the molar absorptivity is 5.36 x 10(4) L x mol(-1) x cm(-1). Beers law was obeyed in the range of 0-40 microg Fe. Iron and eriochrome cyanine R formed a stable complex with a molar ratio of 1:2. The proposed method has been applied to the determination of copper in aluminium alloys with satisfactory results. PMID- 15819023 TI - [Simultaneous determination of molybdenum and chromium by artificial neural networks spectrophotometry]. AB - In this paper, a three-layer artificial neural networks were applied to simultaneous determination of molybdenum and chromium by spectrophotometry. The apparent molar absorptivities of the complex of molybdenum and chromium are 1.4 x 10(5) L x mol(-1) x cm(-1) and 3.7 x 10(4) L x mol(-1) x cm(-1), respectively. A method has been developed for simultaneous determination of molybdenum and chromium in steel with satisfactory results. The paralysis in the procedure of training about ANN has been avoided with the improved back propagation algorithms. The simple transformation method for target vectors and criterion function convenient for selection of the parameters about the networks has been put foward. PMID- 15819024 TI - [Research into determination of tricomponent sample by ridge regression spectrophotometry]. AB - In this paper ridge regression spectrophotometry was used to analyze three components--Acetaminophen, P-Ni-trophenol and P-Aminophenol with absolute ethanol as the solvent. The basic principle and analytical steps of the method were described in detail. The experimental results show that the recovery of each component ranges from 99.24% to 101.31% with the medium of absolute ethanol. As compared with conventional methods, this method is simple, rapid and suitable for computer-aid analysis. PMID- 15819025 TI - [Fluorescence study on phenothiazine and its halogenate derivatives]. AB - The fluorescence spectra of 3-bromine-N-ethyl-phenothiazine with various acid degrees and different polarity surfactants in aqueous solutions were studied in detail. The reasons why fluorescence spectra of Br-Pwere very sensitive to environmental polar and acid changes were discussed based on Lewis acid-base theory. Meanwhile the spectrum of Br-P under ultraviolet ray illumination showed that Br-P had kinetic fluorescence. The method is not only simple but also sensitive with low limit of detection. The calibration curves were linear between 1 x 10(-7) and 1 x 10(-4) mol/L, with a regression coefficient of 0.9995 and RSD<2.8%. PMID- 15819026 TI - [Effect of non-uniform heating of ordinary microwave oven upon the determination results of sample digested in the oven]. AB - This article studies the heating variation of the sample placed on various relative positions of the turning tray in the microwave oven cavity. We find the distribution regularity of heating points that are of insignificant variation statistically, and determine Cu content in the sample GBW07233 and Cu and As contents in the sample NIES SRM No. 1 that are digested on these points. The determinations prove that the precision and accuracy of the determination results will not reduce, even though there are insignificant variations of temperature on these points. PMID- 15819027 TI - [The free searchable patent databases on Internet]. AB - The names and the addresses of the free searchable patent databases on Internet were provided in this paper. The basic ways of information search methods were also introduced. PMID- 15819028 TI - [Femtosecond time-resolved fluorescence spectra of LDS751]. AB - The femtosecond time-resolved fluorescence depletion method has been introduced. In this method two beams of femtosecond laser was used. One excited the sample molecular to emit fluorescence. Then after specified time delay the second laser beam was introduced to deplete the fluorescence through the stimulate emission process. This method has been applied to LDS751 molecules in solution and its time-resolved fluorescence spectrum has been obtained. PMID- 15819029 TI - [A preliminary comparision of the abilities of factor analysis and wavelet analysis in reducing noise of absorption spectra]. AB - In this paper, the abilities of factor analysis and wavelet analysis for reducing noise of the absorption spectra are compared. Also, the different joint ways of these two techniques are used to reduce the noise of the same spectra. Some preliminary results are obtained. PMID- 15819030 TI - [Time-resolved fluorescence investigations of aggregation in Langmuir-Blodgett multilayers of hemicyanine dye]. AB - UV-visible absorption and steady-state and time-resolved fluorescence techniques were used to investigate the aggregation behavior in Y-type hemicyanine Langmuir Blodgett (LB) multilayers. Hemicyanine molecules formed H aggregates in LB films leading to blue-shifts of the absorption and fluorescence bands with respect to those of monomeric hemicyanine in chloroform solutions. The aggregates in LB films were dissociated by mixing with arachidic acid. The measurement of fluorescence decay profiles could be used to quantitatively characterize the lifetime, weights and size of aggregates. PMID- 15819031 TI - [Experimental study of the properties of two-order FADOF at Cs 455 nm line under Nd:YAG pulse laser condition]. AB - This paper experimentally studied some important properties of two-order FADOF at Cs 455 nm line under Nd:YAG pulse laser condition. At a selected temperature the properties of both the bandwidth and the transmission related to the magnetic field have been obtained. In this study, the bandwidth of two-order FADOF was increased linearly with the increment of the magnetic field from 200 to 700 Gauss and is obviously narrower than that of the two single-order FADOFs. The transmission of two-order FADOF was also linearly increased with the increment of the magnetic field in the same range and is lower than that of the single FADOF. Fortunately, at a slightly higher cell temperature, such as T = 160 degrees C, the transmission of two-order FADOF can reach 40%. Furthermore, it can reach theoretically up to 80% because 90% can be obtained in the case of single FADOF. The most important result in the study is that the out-of-bandwidth rejection, which is one of the three parameters, was improved from 10(-3) to 10(-5). The structure of the two-order FADOF is relatively simpler than that of the single order FADOF. PMID- 15819032 TI - [Bonding behavior of 4f orbital]. AB - In this paper, the bonding behavior of 4f orbital in lanthanide coordination compounds was discussed and corresponding theory and research methods were summed up with 36 references. PMID- 15819033 TI - [Calculation for Franck-Condon factors of the two band systems beta(B2pi(r) - X2pi(r)) and gamma(A2sigma+ - X2Pi(r)) of NO in the air at high temperature]. AB - From the nuclear Schrodinger equation of a diatomic molecule with vibration rotation interaction term involved in it, we derive the wavefunction relevant not only to vibrational quantum number, but also to rotational one. The Franck-Condon factors of the two band systems beta(B2pi(r) - X2pi(r)) and gamma(A2sigma+ - X2Pi(r)) of NO in the air at high temperature are calculated in this paper, using the derived wavefunction. In the calculation, the rotational quantum number J takes values from 0 up to 200. The results apply to both low or high temperature conditions. PMID- 15819034 TI - [Microbeam nuclear reaction analysis and its application to individual fluid inclusions]. AB - The principles and analytical technique of nuclear analysis of scanning microprobe (micro-NRA) was introduced. It is a useful tool for nondestructive light elements analysis which can not be detected by micro-PIXE. Using 19F (p, alphagamma)16O reaction at 872 keV resonance energy, the fluorine concentration in trapped fluid inclusions of mantle derived olived-evidence from Cenozoic basaltic rocks in Eastern-China was determined. PMID- 15819035 TI - [Investigation of photoacoustic spectroscopy for polydiacetylene Langmuir Blodgett films (II) synthetical analyse of the spectra and fluorescence quantum efficiency]. AB - In this paper the polydiacetylene (PDA) Langmuir-Blodgett (LB) films, which have potential applications in industry, are studied by using photoacoustic spectrum (PAS) combined with optical absorption and fluorescence spectra. The synthetical analysis of PAS and optical absorption and fluorescence spectra for PDA LB films that underwent environment with different kinds of gases shows that the information about relaxation process of excitation state and feature of elementary excitation can be achieved. Besides, fluorescence quantum efficiency was discussed. The achievment of this paper shows that the PAS technique is a valid means for studying LB films and can capture information which cannot be achieved by other optical spectroscopy. PMID- 15819036 TI - [Improving spectral property of light source to increase signal-to-noise ratio of optical multichannel analyzer in measurement of absorbances]. AB - In the present paper we report that the spectral property of light radiation was improved by using the method of increasing intensity of short wave radiation for increasing signal-to-noise ratio (S/N) of CCD-OMA in measurement of sample's absorbance in visible light-region especially short wave region. The signal-to noise ratio resulting from the recording of short wave region for improved light source was increased by 8 times as high as that of a general Wlamp. As demonstrated by the experiment, absorbance spectrum with high S/N ratio can be obtained in the entire visible light region by using the improved light source in the measurement of absorbance. PMID- 15819037 TI - [FTIR and fluorescence emission spectra of sulfonated PS and its ionomers]. AB - The FTIR spectra of sulfonated polystyrene and Zn and La ionomers change with sulfonation degree and the kind of metal ion. With increasing the sulfonation degree, the stretching vibrational strength of -SO3H and -SO3 increases. Because of the interaction of polar group, the FTIR spectra around 1200 cm(-1) or so split. On the other hand, the degree of sulfonation influences somewhat on the intensity and position of fluorescence spectra of sulfonation polystyrene and its ionomers. PMID- 15819038 TI - [Absorption spectra or hypersensitive transition in the system of neodymium of erbium with amino-acid and 1,10-phenanthroline]. AB - The absorption spectra of f-f hypersensitive transition for the complexes of Nd3+ or Er3+ with some alpha-amino-acids and 1,10-phenanthroline in 95% ethanol solution were studied. The oscillator strength (P) and the spectral parameters (beta, delta, b(1/2)) were calculated. The characteristic of the coordinate bonds of Nd3+ or Er3+ and alpha-amino-acids and the covalent character of the complexes were discussed. The result of the experiment shows that the oscillator strength (P) is related to the numbers of the atoms which form the coordinate bonds. PMID- 15819039 TI - [INDO/CI studies on the electronic spectra of tetraphenylporphyrin and its derivatives]. AB - MNDO calculations and group theory were carried out on tetraphenylporphyrine (TPP) to obtain its optimum molecular geometry. We employed an INDO/CI method to study the electronic structures of tetraphenylporphyrine and its derivatives and identify their main electonic absorption bands. Five-orbital model was advanced to explain the typical characteristics of electronic spectra of H2TPP and its derivatives. The calculated wavelengths were in good agreement with the experimental values. The effects of the substitution of variant group were discussed and some helpful conclusions were drawn. PMID- 15819040 TI - [The polymerization of C60/C70 in air at high temperature]. AB - C60(C70 contained) was sealed in air and heated for 10 minutes at high temperature (850-900 degrees C). It was found from absorption spectrum analysis and SEM observation that the C60(C70) produced polymerization under the action of nitrogen and oxygen. PMID- 15819041 TI - [Study on the interaction of Cu2Zn2SOD with CoCl2, Co(His)n by spectral and magnetic resonance analysis]. AB - Direct interaction of Cu2Zn2SOD with inorganic metal compound (CoCl2) and organic metal compound [Co(II)(His)n] was studied by ICP, VIS, NMR and measurement of enzyme activity. It has been found that in aqueous solution, there exists a direct interaction of the metal ions of the active center in the metalloenzyme (Cu2Zn2SOD) with external added CoCl2, Co(His)n. As a result, part of the metal ions in metalloenzyme were replaced and the corresponding metalloenzyme derivatives were produced and the catalytic activity of enzyme were affected. Moreover, the interaction of Co(II)(His)n with Cu2Zn2SOD is stronger and quicker, than that of CoCl2 with this enzyme, as well as the Co(II) from Co(II)(His)n is easier to enter the active center of enzyme. PMID- 15819042 TI - [An algorithm for real-time quantitative analysis of remote detection spectrum of chemical vapor with passive Fourier-transform infrared spectroscopy]. AB - The algorithm for quantitative analysis of passive Fourier-transform infrared spectum (FTIR) of a chemical vapor is given. It can be applied to both real-time processing for chemical vapor and quantitative measurements of chemical vapor column density. The algorithm simultaneously achieves quantitation of the vapor spectral transmission, background removal, real-time calibration for FTIR. Finally, we discuss in detail an example of the use of the algorithm to a simulant DMMP. PMID- 15819044 TI - [The theoretical study of Raman imaging for pendent drop adsorbed by uniaxial orientated molecules]. AB - In this paper, Raman imaging method used to determine molecular orientation of LB film on pendent drop is proposed. For the molecules which is of uniaxial orientation, all of Raman activity matrix elements have been calculated. The intensity distribution of Raman imaging for pendent drop and the ratio distribution of polarized Raman intensity used to determining molecular orientation have also been calculated and drawn out. PMID- 15819043 TI - [Prediction the octane number of gasoline by near infrared spectra analysis]. AB - The feasibility of predicting the octane number by the near infrared spectra analysis technique in the quantitative analysis was investigated. 12 calibration gasoline samples and 4 prediction samples were collected and their absorption spectra were measured with a high precision model Lambda 19 scanning spectrophotometer. Based on the multivariate statistic model, stagewise regression and partial least squares were selected. A model of the correlation behavior between the spectra and octane number was established and calibrated, and then the prediction of octane number is made. The prediction error is +/-1.0 octane number in the best analysis results. PMID- 15819045 TI - [Research into the mechanism of fragile sealing on the composition packet]. AB - The research into the fragility of the sealing on two composition films for ham packages by using Fourier infrared spectrometer, DTG and melting index detector is reported. The research shows that the cause of the fragility of the sealing is the differences in the materials and their softening points and melting indexes between the inner film and the outer film. The inner film is made of adhesive polypropylene, whose softening point is 148.8 degrees C and melting index (MI) is 6.049 g/10 min. However, the outer film is made of polyamide 610, whose softening point is 221 degrees C, 72.2 degrees C higher than that of inner film, and melting index is 3.09 g/10 min, almost half of the MI of inner film. The mechanism of the fragile sealing on the composition packet is also discussed. PMID- 15819046 TI - [Synthesis and spectroscopic characterization of O,O,O',O'-tetramethyl-S,S'-(2 N,N-dimethylamino-trimethylene)-bis-phosphonodithionate oxalate]. AB - A new organophosphorus insecticide, shacanlin, was synthesized by 1-dimethylamino 2,3-dichloropropane reacting on O,O-dimethyl-dithiophosphate and its oxalate was prepared. The molecular structure of shacanlin oxalate was characterized by 1H and 13C NMR spectrometry, IR and MS. The results indicate that the reaction product is 1,3-disubstituted product. The molecular structure of shacanlin oxalate is (CH3)2N+ HCH[CH2SP(S)(OCH3)2]2 x C2O4H-[O,O,O',O'-tetramethyl-S,S'-(2 N,N-dimethylamino-trimethylene)-bis phosphonodithionate oxalate]. PMID- 15819047 TI - [Study on syntheses and spectroscopic property of complexes of quinoxaline-2,3 dicarboxamide with transition metals]. AB - Three new solid complexes of transition metal Co(II), Ni(II) and Cu(II) with quinoxaline-2,3-dicarboxamide were synthesized, and the composition of the complexes was confirmed to be M(Qxda)2Cl2 [M = Co(II), Ni(II), Cu(II)] by elemental analysis, molar conductance and thermal analysis. The coordination pattern and structure were discussed by infrared spectra, electronic spectra. PMID- 15819048 TI - [The Raman spectra of rubidium titanyl arsenate]. AB - The Raman spectra of rubidium titanyl arsenate (RTA) single crystal with different geometric arrangements were obtained and compared with those of rubidium titanyl phosphate (RTP), potassium titanyl arsenate (KTA) and potassium titanyl phosphate (KTP) single crystals. The assignments of various vibration modes were made according to the experimental results. The strong Raman scattering intensity implies that RTA crystal possesses large nonlinear optical property and the strongest scattering is from the TiO6 octahedron. From the mode splitting of AsO4 tetrahedron, we conclude that the distortion of AsO4 in RTA is larger than that of PO4 in KTP. PMID- 15819049 TI - [Synthesis and spectroscopic characterization of complexes of Y(III) and Nd(III)]. AB - The complexes of Y(III) and Nd(III) with bis-Schiff bases from 2-hydroxy-3 methoxy benzaldehyde and benzidine were synthesized and characterized by elemental analysis, molar conductivity, IR, UV and 1H NMR. The possible structure and some properties were studied. PMID- 15819050 TI - [Analysis of odoriferous pollutants in cooling water by on-line LC-GC/FTIR method]. AB - The integration of two dimensional chromatography system (LC-GC) has ability for clean-up, enrichment, pre-separation and high resolution separation for the sample. The coupling of LC-GC with FTIR can notably improve the detection ability of conventional (light pipe) GC/FTIR system. The interface of LC-GC system employs retention gap technique (15 m x 0.53 mm i.d. retention gap column and 4 m x 0.25 mm i.d. precolumn) with on-column injection and partially concurrent solvent evaporation with early vapor exit. The applicability of the method is demonstrated by the analysis of odoriferous pollutants (phenolic compounds) in the cooling water with 15 cm x 2 mm i.d. silica LC column and 35 m x 0.25 mm i.d. OV-101GC column under the conditions of haxane/dichloromethane (80:20) as the mobile phase (0.3 mL/min) of LC, N2 as the carrier gas in GC and solvent evaporation temperature 80 degrees C, inlet pressure of 0.2 MPa etc. The method of on-line coupled LC-GC/FTIR is a reliable analytical tool for microcomponents in complex mixtures. PMID- 15819051 TI - [Interatomic distances of molecule for orthohydrogen and parahydrogen]. AB - The determination of rotational Raman spectra for hydrogen gas at 354.9, 587.4, 814.7 and 1035.1 cm(-1) by FT-Raman spectrometry is described in this paper. The Raman bands at 354.9 and 814.7 cm(-1) were assigned to rotational levels J = 0 - > J = 2 and J = 2 --> J = 4 for parahydrogen, and those at 587.4 and 1035.1 cm( 1) were assigned to rotational levels J = 1 --> J = 3 and J = 3 --> J = 5 for orthohydrogen. The interatomic distances determined for orthohydrogen and parahydrogen molecules with nonrigid rotator model are 75.03 and 74.98 pm, respectively. PMID- 15819052 TI - [Analysis of Chinese moxibustion in visible and infrared spectra in process of clinical application]. AB - By use of the testing compartment which was made up and the spectrometer of visible or infrared light give expression to electromagnetic spectrum of that the heat is given by Chinese moxibustion in process of clinical application. Experimental results show that the heat wich is given when Chinese moxibustion is burnt is the continuous wave electromagnetic spectrum from about 600 millimicron to far infrared and diffrent electromagnetic spectrum are given out by diffrent kinds of Chinese moxibustion. PMID- 15819053 TI - [Hyphenation of capillary electrophoresis with nuclear magnetic resonance and its applications]. AB - Direct on-line coupling between capillary electrophoresis and nuclear magnetic resonance spectroscopy (CE-NMR) is one of the most powerful and novel techniques for simultaneous separation and structural elucidation of biomacromolecules. Recent advances and the most crucial technique of measuring samples on nanoliter scale in CE-NMR are described in detail. PMID- 15819054 TI - [Development and evaluation of a system for pulse generation and signal processing control in a plasma atomic fluorescence spectrometer with hollow cathode lamp as excitation source]. AB - A Z80 processor was replaced with a 386 microcomputer to control pulse generation for hollow cathode lamp and fluorescence signal processing in a Baird Plasma AFS 2000 atomic fluorescence spectrometer. An interface board was designed for control purpose. The system was evaluated with detection limits, relative standard deviations and linear regression coefficients of 15 elements, which were in good agreement with the corresponding values in the manufacturer's manual. With the higher-level microprocessor in the system, the performance of the instrument was improved. PMID- 15819055 TI - [Atomic/ionic fluorescence in microwave plasma torch discharge excited by high current microsecond pulsed hollow cathode lamp-europium atomic/ionic fluorescence spectrometry]. AB - Eu atomic and ionic fluorescence spectrometry in microwave plasma torch discharge excited by high current microsecond pulsed hollow cathode lamp (HCMP HCL-MPT AFS/IFS) was studied. Operating conditions were optimized. The best detection limits for AFS and IFS obtained with a desolvated ultrasonic nebulization system were 42.0 ng/mL for Eu I 462.7 nm and 21.8 ng/mL for Eu II 381.97 nm, respectively, both were better than those given by the instruction manual of a Baird ICP AFS-2000 spectrometer using pneumatic concentric nebulizer with desolvation for AFS, but were significantly higher than those obtained by using the Baird spectrometer with a mini-monochromator and a ultrasonic nebulzer system. PMID- 15819056 TI - [Determination of electron density in Ar-air mixed coolant gas ICP]. AB - The lateral distribution of electron number density at 0, 2, 5, 10 mm above the load coil in Ar-air mixed coolant gas ICP was measured with Stark broadening method of Hbeta line (486.1 nm) and some of the lateral distribution was converted into radial distribution by Abel inversion under the condition similar to that in the low power Ar-ICP. The results show that the distribution of electron number in Ar-air mixed coolant gas ICP is similar to that in Ar-ICP and exhibit off axis maxima. When a lot of air was added to the coolant, the positions of the off axis maxima would near the axis. The electron number density reached maximum and was higher than that in Air-ICP as 5%-10% air was added to the coolant. PMID- 15819057 TI - [A comparison of Fe, Zn, Cu, Cr, Cd and Pb concentration in serum determined by ICP-AES using different sample preparation procedures]. AB - Fe, Zn, Cu, Cr, Cd and Pb concentrations in serum of 21 arteriosclerosis (AS) and 18 healthy control (CK) were determined by ICP-AES, and sample solution was prepared using acid digestion (Dig.) and acid dilution (Dil.) procedure. Serum total Fe, Zn and Cr were significantly higher using Dig. than that using Dil. Serum Cu and Pb were of no significant differences using the both procedures. Serum Cd was significantly lower using Dig. than that using Dil. The results of Fe, Cu, Zn and Cr were positively correlated between Dig. and Dil., but those of Pb and Cd were of no significant correlation. Six elements concentrations in serum determined were significantly different between AS and CK using Dig. The digestion procedure was recommended for serum trace elements analysis using ICP AES. PMID- 15819058 TI - [Determination of Ca, Cu, Zn, Mg, Fe, P, Ni, Mn, Mo and Ba in the secretion of Psylla Chinese Yang et Li by ICP-AES]. AB - A method of simultaneous and direct determination of Ca, Cu, Zn, Mg, Fe, P, Ni, Mn, Mo and Ba in secretion of Psylla Chinese Yang et Li by ICP-AES is reported. The results show that the method has good precision and accuracy. The relative standard deviation is less than 5.8%. The recovery is 94%-106%. This method is also simple and rapid. PMID- 15819059 TI - [Comparative studies on five pretreatment methods in the determination of elements in plant standard sample by ICP-AES]. AB - Direct determination of the contents of 19 major or trace elements: K, Na, Ca, Mg, Si, Al, Fe, P, Sr, Ti, Mn, Zn, Pb, B, Ba, Cu, Li, V and Ni in plant samples by ICP-AES are reported in this paper. The solution of plant standard sample GBW07603 was prepared by five pretreatment methods: dry-ashing, HNO3 + H2SO4 + HClO4, H2SO4 + H2O, HNO3 + H2SO4 + HClO4 + HF and ashing + NaOH method. Suitable pretreatment methods were found for the determination of the elements by comparing standard deviations and relative errors of the results, thus providing a scientific basis for the selection of suitable pretreatment of chemical element analysis in plants. PMID- 15819060 TI - [Determination of Si, P, Nb and Ti in steels by ICP-AES with offset profile additional lines]. AB - This article introduced the increment of measurement channels for the elements silicon, phosphorus, niobium and titanium by changing the entrance slit angle, based on the fixed channels of the ICP-AES. The entrance slit angle was controlled by a motor. We determined trace silicon, phosphorus, niobium and titanium in steels, and examined the detection limits, accuracy and precision, and the results were satisfactory. With this method, we could also determine other elements, and widened the applied range of ICP-AES. PMID- 15819061 TI - [Study on the determination of iodine by ICP-AES]. AB - A method for the determination of iodine in iodo-calcium pieces by ICP-AES was studied. I- was turned to I2 by HClO4 and H2O2. The detection limit of iodine is 1 ng/mL. The sample was dissolved with water and was directly determined without seperating the precipitate in the bottom of volume flask. It was found that the precipitate attracted with I2 and the quantity of attracting would increase with delay time. PMID- 15819062 TI - [Flow injection analysis of 3,4-dihydroxybenzoic acid with inhibited chemiluminescent detection]. AB - It was found that 3,4-dihydroxybenzoic acid (protocatechuric acid) inhibited chemiluminescence of luminol-H2O2 reaction catalyzed by Co2+. On this basis, a flow injection method has been developed for the determination of protocatechuric acid. The method is simple, convenient and sensitive with detection limit of 2.7 x 10(-7) mol/L and is effective to determine protocatechuric acid in the range of 1 x 10(-6)-1 x 10(-5) mol/L. The variation coefficient of eleven determinations for 8 x 10(-6) mol/L protocatechuric acid is 2.3%. PMID- 15819063 TI - [Background absorption and its correction in the determination of Co in steel with flame atomic absorption spectrometry]. AB - In this paper, a study of background absorption and interference of Fe in the determination of Co with flame atomic absorption spectrometry was done. The influence of background absorption on Co 240.7 nm could be completely eliminated when Au 242.8 nm was used as a background correction line, and the results of the determination of micro-amount Co in iron and high-alloy steel with standard addition method were satisfactory. The mechanism of background absorption and interference was also discussed. PMID- 15819064 TI - [Determination of serum and erythrocyte iron, serum ferritin, MCH and MCHC related with aged hypertension and coronary heart disease]. AB - Using AAS method, we detected serum and erythrocyte iron, serum ferritin (SF), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) in the elders with hypertension and coronary heart disease. The results showed that the levels of serum iron, erythrocyte iron and SF were significantly higher than those in the healthy control (P < 0.01). It is concluded that iron metabolic abnormality may be one of the factors resulting in hypertension and coronary heart disease. PMID- 15819065 TI - [Determination of metals in atmospheric particulates by atomic absorption spectrometric method (AAS) with CA-CN micropore membrane filters used for sampling]. AB - A new sampling method of atmospheric particulates using 0.22 microm pore diameter micropore membrane filters was introduced. Copper, lead, zinc cadmium, manganese, iron, nickel and chromium in atmospheric particulates were determined by atomic absorption spectrometry. This sampling method has shown several advantages: easy, complete and fast digestion; low membrane blank; increased flexibility to surrounding temperature change; high elution recovery, standard addition recovery, precision and accuracy. PMID- 15819066 TI - [GFAAS determination of trace lead in toothpaste using L'vov platform]. AB - Sensitivity in determining low melting point metals is highly improved by means of platform technique in graphite furnace AAS. In this paper we describe a method of determining trace lead in toothpaste by this technique. The method of making the platform and the selection of the platform coating were discussed and the results with and without the platform were compared. The optimum ashing temperature, atomization temperature and experimental conditions were investigated. The interference of large amount Al and Ca in the matrix on determination of trace lead can be minimized by using platform technique. The precision of about 0.9% and detection limit of about 15 ng obtained by this improved method are much better than those by methods without platform. PMID- 15819067 TI - [Determination of As, Hg and Mo in niuhuang jiedu tablets and other Chinese traditional medicines with graphite furnace AAS]. AB - The determination of As, Hg and Mo in the composite Chinese traditional medicines such as niuhuang jiedu tablets etc. Indicates the existence of As, Hg and Mo in these medicines. The relation between these elements and the efficacy of the medicines was studied, showing that these elements play an important role in the efficacy of the drugs. PMID- 15819068 TI - [Determination of mercury in food by microwave oven digestion-hydride generation atomic absorption spectrometry]. AB - Mercury in food was determined by microwave oven digestion-hydride generation atomic absorption spectrometry. The best condition of digestion was also studied. Furthermore, the way of doing digestion was compared to standard method with satisfactory results. The method is rapid and simple, and of good precision and high accuracy. The sensitivity is 0.43 microg/L. The detection limit is 0.35 microg/L. The relative standard deviation is 2.8% and the recovery is 93.5% 103.0%. PMID- 15819069 TI - [Determination of selenium in human hair with graphite furnace atomic absorption spectrometry]. AB - Determination of selenium in human hair by GFAAS has been described in this paper, Hairs from 120 middle school teachers were determined. The results are 0.15-1.00 mg/kg and the median is 0.43 mg/kg. The sensitivity of this method is 2.96 ng/mL and the relative standard deviation is 2.4%-6.7%. PMID- 15819070 TI - [Experimental study on the mechanism of chemical interference of tellurium with selenium by hydride-AFS in the presence of Fe3+]. AB - This paper describes the coaction of tellurium and Fe3+ on selenium in hydrogenization by experiment. The experimental result proved that certain amount of Fe3+ can eliminate the interference. Under optimum conditions, the reaction mechanism was discussed. PMID- 15819071 TI - [Study on the relationship between the concentration of iron manganese and turbid degree in river by AAS]. AB - The concentration of Fe, Mn and the turbid degree at different section in the head waters of Fen River were determined by AAS and turbid meter. A quantitative model parameters were obtained based on the treatment of mathematical and physical statistics. The relationship between the concentration of Fe, Mn and turbid degree was established exactly. PMID- 15819072 TI - [Determination of zinc in vegetable oils by FAAS with trace injection method]. AB - A method for the determination of zinc in the soybean, cottonseed and sesame oils by FAAS with trace injection technique was discribed in this paper. The injection volume was 30 microL. The characteristic concentration and the detection limit were 0.098 and 0.00368 microg/g, respectively. The relative standard deviation was 2.6% and the recoveries were in the range of 92.9%-107.8%. The contents of zinc in the samples were 9.67, 1.42 and 1.77 microg/g, respectively. PMID- 15819073 TI - [Determination of potassium, sodium, chromium and nickel in 11 Chinese traditional medicines by FAAS]. AB - Four metallic elements in 11 Chinese traditional medicines such as potassium, sodium, chromium and nickel were determined by flame atomic absorption spectrophotometer. The results showed that there are rich metal elements, but the contents are different in 11 Chinese traditional medicines, providing useful data for discussing relations between trace elements in Chinese traditional medicines and the cure for CHD. PMID- 15819074 TI - [Continuous determination of Zn, Cu and Mn in tea by flame atomic absorption spectrophotometry]. AB - This paper reports the determination of trace metals Zn, Cu and Mn in tea by flame atomic absorption spectrophotometry. The method is simple and rapid with good precision and accuracy. The characteristic concentration is 0.009 microg/mL/1% for Zn, 0.031 microg/mL/1% for Cu and 0.017 microg/mL/1% for Mn, respectively. The relative standard deviation is between 0.9% and 5.0%. The recovery rate is between 92.0% and 106.0%. PMID- 15819075 TI - [Study on the determination of aluminum in soil samples by air-acetylene flame atomic absorption spectrometry]. AB - This paper reported a method for the direct determination of aluminum in soil samples by air-acetylene flame atomic absorption spectrometry. The enhancement effect on aluminum of adding tetramethylammonium chloride was studied. 43 microg/mL/1% of characteristic concentration was reached. This method is simple, rapid and useful, and has been applied to the determination of aluminum in standard soil samples, and a good agreement was achieved. PMID- 15819076 TI - [Determination of Cu, Zn and Fe in different muscle tissues of pigs and chickens by FAAS]. AB - The contents of Cu, Zn and Fe in different muscle tissues of pigs and chickens were determined by flame atomic absorption spectrophotometry (FAAS). Mixed acid of HNO3 and HClO4 was used to digest wet samples of the tissues. A comparative analysis of sample preparations was carried out in this paper. PMID- 15819077 TI - [Determination of 11 metal elements in niuhuang jiangya wan by atomic absorption spectrophotometer]. AB - Eleven metal elements in niuhuang jiangya wan, including sodium, calcium, magnesium, potassium, iron, chromium, manganese, nickel, cadmium, copper and zinc, were determined by flame atomic absorption spectrophotometer. The results showed that there are rich metal elements in niuhuang jiangya wan and provided useful data for discussing the relation between trace elemtnts in niuhuang jiangya wan and cure for high blood pressure. PMID- 15819078 TI - [Direct determination of zinc and iron in maize flour using suspension sample introduction by flame atomic absorption spectrometry]. AB - A new FAAS method for the determination of zinc and iron in maize flour using suspension sample introduction is described. The maize flour was suspended in agar sol, and injected into air-acetylene flame by using standard addition method to determine zinc and iron. The results are consistent with those obtained by ashing method. This method is convenient and rapid. PMID- 15819079 TI - [Indirect determination of trace silicon in cobaltosic oxide by extraction-FAAS]. AB - After the heteropolymobdate of silicon and phosphorus was formed, the heteropolymobdate of phosphorus was extracted into CH3COOH4H9 phase and P as interference element was separated. The heteropolymobdate of silicon was extracted by MIBK. Indirect determination of Si was achieved with Mo in MIBK determined by FAAS. Applying this method to the determination of silicon in cobaltosic oxide, the interferences of matrix cobalt and other coexistence elements were avoided. Compared to spectrophotometric method, the result is satisfactory. PMID- 15819080 TI - [Determination of calcium and magnesium in wheat flour by suspension sampling flame atomic absorption spectrometry]. AB - Suspension sampling technique was applied to flame atomic absorption spectrometry and was successfully used to determine calcium and magnesium in wheat flour. The wheat flour was suspended in agar sol containing dibutyl phthalate and made into suspension. Choice of suspension agent and elimination of chemical interference were studied. The test solution was injected into air-acetylene flame to determine calcium and magnesium by standard addition method. Determination results were consistent with those obtained by ashing method. The t-test showed that no difference was found between the two methods. Displacement of ashing method by suspension sampling method for the sample pretreatment is possible. This method is convenient, rapid and accurate. PMID- 15819081 TI - [Neural cluster structure with single component prediction in multiple variable systems for X-ray fluorescence spectrometry]. AB - A neural cluster structure with single component prediction (NCSCP) was proposed for X-ray fluorescence spectrometry in a multivariable system. The neural cluster structure is built by the collection of a group of neurons which have close relationships among one another. In X-ray fluorescence analysis, the structure is constructed by choosing the elements in which there exist serious matrix effects, and deleting the components containing large noise. The predictability of the neural cluster structure was compared with that of the classical backward error propagation algorithm with single component prediction. The results show that the neural cluster structure is significantly superior to the classical algorithm in prediction accuracy, antidisturbance and the predictabilty to outliers. PMID- 15819082 TI - [TXRF technique and quantitative analysis of mollusc teeth]. AB - Total reflection X-ray fluorescence (TXRF) analysis technique and the instrument with a short path, high efficiency, low power and small volume are briefly presented. The detection limit of the system are at pg-level for Cu and Mo target excitation. Teeth of a marine mollusc were measured quantitatively and the spectrum and analysis results were given. PMID- 15819083 TI - [X-ray diffraction spectrum of heroin]. AB - In this paper, practical measured X-ray diffraction spectra of heroin and opium are given and the parameters of each diffraction peak of the heroin are listed. The heroin belongs to orthorhombic crystal system; the basic vectors of the primitive cell are: a = 8.003, b = 14.373, c = 16.092 x 10(-10) m. As compared with the standard spectra of pure heroin and sucrose, the main doped additive checked by us, is sugar affirmatively. PMID- 15819085 TI - [Determination of P in superhigh-P cast iron by EDXRF]. AB - In this paper preparation of standard samples of superhigh-P cast iron has been studied and P in the samples has been determined by EDXRF and powder compacting. In the process the procedure of powder compacting and the software of a multifunctional Lucas-Tooth intenstity correction have been developed. The method is rapid and simple. Analytical results were satisfactory. PMID- 15819084 TI - [Analysis of carbon in low alloy steel by XRF]. AB - In this paper optimum conditions for the analysis of carbon have been found and a new sampling method introduced. The detection limit is 0.0179% and the standard deviation is 0.0039% as C=0.18%. The method has been used for the determination of carbon in low alloy steel with satisfactory results. PMID- 15819086 TI - [Determination of protein in urine by spectral probe of molybdenum(VI) salicylfluoronate complex]. AB - The absorption spectral behavior of binding reaction of molybdenum(VI) salicylfluoronate on bovine serum albumin was investigated. A novel method for the determination of protein in urine using Mo(VI)-salicylfluoronate complex as a spectroprobe was developed. The maximum binding number (n) of molybdenum complex to BSA was found to be 100. The molar absorptivity of binding product at 575 nm was 2.86 x 10(6) L x mol(-1) x cm(-1). The method possesses high sensitivity as well as high selectivity. It could be used to determine protein in human urine directly. The relative standard deviation for the determination is 1.5%. PMID- 15819087 TI - [Study of the fluorescence of terbium-pipemidic acid system and the determination of pipemidic acid in urine and serum]. AB - Intramolecular energy transfer occurs in the terbium complex with pipemidic acid (PPA) and the terbium(III) ion fluorescence can be sensitized by PPA. The fluorescence intensity of the Tb3+-PPA system is proportional to the amount of PPA. Accordingly, a fluorimetric method for the determination of PPA is proposed. The characteristics of the method include low background noise, high selectivity and long fluorescence lifetime. The method can be applied to the determination of PPA in urine and serum. The recovery is in the range of 94.6%-102%. Its detection limit and linear range for PPA are 3.6 ng/mL and 5.0 x 10(-8)-4.0 x 10(-6) mol/L, respectively. PMID- 15819088 TI - [Synthesis and characterization of lanthanum(III) with O-chlorophenoxyacetic acid]. AB - This work has synthesized a binary solid complex using rare earth ion La3+ with the ligand HL (HL = O-chlorophenoxyacetic acid). The chemical formula of the complex was determined to be LaL3 x H2O by elemental analysis. The composition, structure and properties of this type of complex were tentatiovely investigated by the analysis of molar conductivity, IR, UV, TG-DTA and 1H NMR. The center ion in the complex was considered to be chelated by a bidentate ligand HL. PMID- 15819089 TI - [Fluorescence study on the interaction of salicylic acid and human serum albumin]. AB - The change in fluorescence of human serum albumin (HSA) and salicylic acid due to their interaction has been studied in 0.05 mol/L, pH 9.50 borate buffer (contains 0.01 mol/L KCl). The dissociation constant of salicylic acid from HSA was obtained by using two different methods. PMID- 15819090 TI - [Sensitizing effect of nonionic microemulsion on phosphorus in crystal violet heteropoly molybdophosphoric acid]. AB - A highly sensitive and convenient spectrophotometric method for phosphorus was developed. The sensitizing effect of nonionic microemulsion Triton X-100/n C5H11OH/n-C0H20/H2O on the ion-association of heteropoly molybdophosphoric acid with crystal violet was higher than that of micelle Triton X-100/H2O. This result showed, by the measurement of distribution constant KD, that the binding constant K and the number of binding sites n of CV were related to the greater solubilization capacity of CV-PMo in nonionic microemulsion system than that in micelle. PMID- 15819091 TI - [Fluorescence enhancement of the dysprosium-sulfosalicylic acid system by lanthanum and its application]. AB - A fluorescence enhancement produced by adding lanthanum(III) to dysprosium sulfosalicylic acid system was observed. La3+ enhanced the fluorescence intensity by two orders of magnitude compared with the system without La3+. The system was used for the determination of trace amount of dysprosium in the range of 3.0 x 10(-7)-1.0 x 10(-5) mol/dm3, and the detection limit was 8.0 x 10(-8) mol/dm3. The procedure was applied to the determination of dysprosium in synthetic rare earth samples and standard rare earth samples with satisfactory results. PMID- 15819092 TI - [Columinescence effect of dysprosium-lanthanum-tiron system and its applicaton]. AB - The columinescence effect of dysprosium-Tiron system by lanthanum was discovered and the characteristics of fluorescence spectra of the system were studied. The effect of the experimental conditions on the fluorescence intensity was defined. By adding suitable amount of lanthanum to dysprosium-Tiron system, the fluorescence intensity can be enhanced by about 40 times. The fluorescence intensity is a linear function of the concentration of dysprosium in the range of 5.0 x 10(-8)-9.0 x 10(-6) mol/L. The detection limit is 1.0 x 10(-10) mol/L. Standard addition method is used for the determination of dysprosium in synthetic rare earth sample with satisfactory results. PMID- 15819093 TI - [Flow injection analysis for trace phosphorus(V) and arsenic(V) determination]. AB - A flow injection analysis method for trace amount P(V) and As(V) determination has been established based on the fluorescence quenching effect of rhodamine 6G due to the formation of an unfluorescence ion-association complex. Rhodamine 6G has a maximum emission wavelength of 555 nm (excitation at 350 nm). The calibration curves were found linear in the range of 0-80 microg/L of P(V) and 0 100 microg/L of As(V). The method has been used for the determination of P(V) and As(V) in copper alloys with satisfactory results. PMID- 15819094 TI - [New solution of simultaneous equations in spectrophotometry for the determination of aspirin and acetaminophen in xiaoer tuishao pian]. AB - The contents of aspirin and acetaminophen in xiaoer tuishao pian were determined by using the new solution of simultaneous equations without any preliminary separation. The average recoveries and variation coefficients of the analysis are 99.95% and 1.74% for aspirin and 100.1% and 0.93% for acetaminophen, respectively. The method was simple, rapid and accurate. PMID- 15819095 TI - [Determination of copper in waste water by beta-correction new spectrophotometry]. AB - In pH 5-6 solution copper [Cu(II)] can react on p-dimethylaminobenzylidene rodamine (PDR) to form red compound. Beta-correction spectrophotometry was studied to determine trace copper. It can remove the absorption interference of surplus PDR in reaction solution to raise the analytical sensitivity. Experiments for samples showed that RSD = 2.9% and the recovery rate was 87.7%-112%. The minimium concentration of the determination is 0.55 microg/25 mL. The method is suited to analyse copper in metals and waste water. PMID- 15819096 TI - [Spectrophotometric determination of trace amount of cadmium in marine products]. AB - The colour reaction of Cd(II) on xylend orange in the presence of hexamethylenetetramine-KNO3-HNO3 has been studied. Cd(II) forms the complex at pH = 6.2-6.4 (lambda(max) = 578.4 nm) with an apparent molar absorptivity of 1.6 x 10(4) L x mol(-1) x cm(-1). Beer's law is obeyed for Cd(II) in the range of 0.5-6 microg x mL(-1). The detection limit is 0.007 microg x mL(-1) with the recovery of 90%-105%. The method is simple and highly sensitive, and has been applied to the analysis of sample with satisfactory results. PMID- 15819097 TI - [Assay of diclofenac sodium and codeine phosphate in tablet by multiwavelength K factor spectrophotometry]. AB - Assay of diclofenac sodium and codeine phosphate in tablet is reported in this paper. The calibration curves were linear over the range of 11.18-25.40 microg/mL of diclofenac sodium and over the range of 6.785-15.42 microg/mL of codeine phosphate. The average recoveries and relative standard deviations of diclofenac sodium and codeine phosphate were 100.1%, 0.36% and 100.7%, 0.80%, respectively. The method is simple and rapid, and the results are satisfactory. PMID- 15819098 TI - [Spectrophotometric assay of polyethyene glycol in urine using ammonium ferrothiocyanate aqueous-chloroform system]. AB - A spectrophotometric method for the quantitative determination of polyethylene glycol-600 (PEG-600) in urine is based on partitioning a chromophore present in ammonium ferrothiocyanate reagent into an aqueous phase and a chloroform phase in the presence of PEG. The method is simple and reproducible, and can detect PEG in a concentration of 0.25 g/L. It gives a linear response over a range of 0.25-2.5 g/L. The recovery of PEG-600 in urine is 107% and relative standard deviation is less than 5%. PMID- 15819099 TI - [Determination and analysis of physical factors affecting insulin fluorescent spectrum]. AB - Insulin is an important factor which regulates cell metabolism and growth. The influences on insulin fluorescent spectrum by pulse electrical field, concentration and temperature were determined in this paper. It was proved that pulse electrical field, concentration and temperature could influence insulin fluorescent spectrum, and thus influence insulin confirmation and insulin biological activity. The mechanism of insulin influenced by pulse electrical field is different to temperature. It is possible that the electrical field influences the insulin fluorescent spectrum by changing Tyr position and the temperature influences the insulin fluorescent spectrum by molecule thermal motion. The insulin confirmation influenced by electrical field is steady. PMID- 15819100 TI - [Synthesis, spectroscopic property and fluorescence of ternary complex of Eu(III) with nicotinic acid and phenanthroline]. AB - The new complex Eu(Phen)L3 of rare earth element Eu(III) with nicotinic acid (abb. HL) and 1,10-phenanthroline (abb. Phen) was synthesized and then characterized by elemental analysis, IR, UV, 1H NMR, fluorescence spectra, X-ray powder analysis, molar conductance, TG and DTA. PMID- 15819101 TI - [Floatation spectrophotometric determination of metals with isochromatic dye ion pairs tetrabromofluorescein-rhodamine 6G system]. AB - The floatation spectrophotometric determination of germanium, platinum and ferrum with isochromatic dye ion-pairs formed by tetrabromofluorescein (TBF) and rhodamine 6G (R6G) is described. The association complexes of these metallic ions with rhodamine 6G floated with organic solvent were first allowed to react on basicity, and then on tetrabromofluorescein which is of the same color as rhodamine 6G. The isochromatic dye ion-pairs of R6G x TBF were floated with toluene. The floatation R6G x TBF in acetone was dissolved and the absorbance of the solution at 530 nm was measured. The sensitivity is raised because of the 2 m dyes. The molar absorptivities for Ge, Pt and Fe are greater than 10(5) L mol(-1) cm(-1). PMID- 15819102 TI - [Study on the double fluorescers fluorometry and its application]. AB - Butylrhodamine B (BRB) was produced and entered into aqueous phase when pH=8.7 buffer solution was added to toluene extractive solution of GaCl4BRB+. And then BRB2 x TBF was extractred by tetrabromofluorescein (TBF). N-butanol (toluene:n butanol 1:1) was added to toluene extractive solution of BRB2 x TBF. The sensitivity was raised because of the assistant effect of BRB2 x TBF. The detection limit was 0.1 microg/L. Beer's law was obeyed in the range of 0-0.15 mg/L with a relative standard deviation of 3.9% for 0.0027% gallium in metal and the recoveries were 94.7%-105.2%. PMID- 15819103 TI - [Indirect determination of industrial chlordane by ultraviolet spectrometry]. AB - Toxicity in chlordane is in proportion to the content of hexachlorocyclopentadiane. Choosing acetone as the solvent, we determined that content by ultraviolet spectrometry, and then calculated roughly the content of this industrial chlordane. The method is cheaper, more rapid than gas chromatography, and suited to rapid testing industrial chlordane. PMID- 15819104 TI - [The thermodynamic molar absorptivity of coordination color reaction of indium(III) with 4,5-dihydroxybenzene-1,3-disufornic acid]. AB - The thermodynamic molar absorptivity of complex formed by 1:1 coordination color reaction of tiron with highly charged cation In3+ has been measured spectrophotometrically at 20.0+/-0.1 degrees C, using NaClO4 as supporting electrolyte. The influence of a on linear relation between molar absorptivity and ionic strength has been investigated. At a = 6.5, the thermodynamic molar absorptivity lg epsilon(o) was found to be 3.69. PMID- 15819105 TI - [Study on the catalytic spectrophotometric determination of trace manganese]. AB - A new catalytic spectrophotometric method for determining trace amounts of manganese has been developed. The method is based on catalytic oxidation of morin by hydrogen peroxide. Manganese has catalytic effect. The detection limit for manganese is 0.004 microg/mL. The linear range of the determination is 5-80 ng/mL of manganese. The method has been used to determine trace manganese in wine. PMID- 15819106 TI - [Simultaneous determination of tungsten, molybdenum and titanium by k-factor dual wavelength standard addition method]. AB - In this paper, a new method for simultaneous spectropthotometric determination of tungsten, molybdenum and titanium is proposed. The method is based on the difference of absorption spectra of the three elements forming coloured complexes with PF in acidic medium and in the presence of nonionic surfactant Tween-80. The absorbance of sample was measured at wavelengths of 503, 530 and 560 nm, and the data were treated by the principle of the k coefficients and dual-wavelength standard addition method. The method has been applied to the determination of tungsten, molybdenum and titanium in steels, and satisfactory results were obtained. PMID- 15819107 TI - [Study on the new determination of analginum]. AB - Analginum in medicine was determined by a new photometric method based on the color formation of Fe(II)-alpha,alpha'-dipyridine complex from alpha,alpha' dipyridine and Fe (III) which could be reduced by analginum. The method was simple and rapid, and the results obtained were in agreement with those of iodine titration method and ultraviolet photometric method. PMID- 15819108 TI - [Determination of ascorbic acid in medicine by sulfosalicylic acid spectrophotometry]. AB - Ascorbic acid in medicine was determined by fading spectrophotometric method, based on Fe3+ reduced into Fe2+ by ascorbic acid and reacting on sulfosalicylic acid. The results obtained were in agreement with those of Chinese pharmacopoeia's method and background correction method. PMID- 15819109 TI - Ensuring ethical standards in an international scientific journal: an evolutionary process. PMID- 15819110 TI - Community Dental Health: the road to excellence. PMID- 15819111 TI - Comparison of a generic and a specific measure of oral health related quality of life. AB - OBJECTIVES: The aims of the study were to compare the discriminant validity of a generic health state measure, the EuroQol, and a specific oral health measure, the Oral Health Impact Profile. PARTICIPANTS: Patients treated by a random sample of South Australian dentists. BASIC RESEARCH DESIGN: Data were collected in 2001 02 using mailed self-complete questionnaires. Dentists recorded the diagnosis of dental conditions and provided patients with self-complete questionnaires to record the nature, severity and duration of symptoms using the EuroQol (EQ-5D+) and 14-item version of the Oral Health Impact Profile (OHIP-14) instruments. Data were available from 375 patients (response rate = 72%). MAIN OUTCOME MEASURES: Three measures (simple counts, additive scores and scale scores) were used for each instrument and compared by oral health, patient demographic and visit factors, and main presenting dental condition. RESULTS: Across all three measures the OHIP-14 was associated with the oral health factors of dentate status, number of teeth, denture wearing and number of decayed teeth while the EuroQol was associated with decayed teeth. For patient demographics both the EuroQol and OHIP 14 were associated with age of patient. For visit factors both the EuroQol and OHIP-14 were associated with visit type and insurance status, while only the EuroQol was associated with time since last visit. Both the EuroQol and OHIP-14 were associated with main dental condition. CONCLUSIONS: There was little difference in the number of associations of either EuroQol or OHIP-14 scores regardless of whether simple counts, additive scores or scale score measures were used. In bivariate analyses the OHIP-14 was more sensitive to oral health factors, but the EuroQol performed as well or better than the OHIP-14 for patient demographics, visit factors and main dental condition, and was associated with the oral health factor of number of decayed teeth. Both measures performed similarly in multivariate analysis. PMID- 15819112 TI - Relationship between oral Candida species and risk factors with reference to drugs with xerostomic side effects among institutionalised elderly in Aichi Pref., Japan. AB - OBJECTIVE: To clarify the relationship between Candida carriage and drugs which have reported xerostomic side effects in the elderly. DESIGN: Cross-sectional study. Setting Two long-term care facilities in Aichi Pref., Japan. PARTICIPANTS: Eight men and 63 women with a mean age of 86 years (range 62 to 95 years) with at least some of their natural teeth. METHOD: Oral examination by dentists, microbiological test by microbiologists, questionnaire interviews, and data obtained from medical files. The Candida species (CFU) was adopted as an objective variable with risk indicators being age, number of teeth, saliva flow rate, denture wearing, xerostomic drugs, activities of daily living (ADL), frequency of brushing and type of meals. RESULTS: Bivariate analysis confirmed that participants with a higher number of Candida species (CFU) fell into the following categories: denture wearing (p < 0.05), older age (p < 0.05), xerostomic drugs (p < 0.10), more teeth, poor ADL, soft/liquid meals, and brushing once a day. This was in contrast to the categories of no denture wearing, younger age, no xerostomic drugs, fewer teeth, good ADL, normal meals, and brushing two or more times a day, in that order of significant probability on an ascending scale. A multiple logistics regression model confirmed that the variables of older age (80 years and over) and more teeth (six or more) had statistically significant (p < 0.05) effects on the number of Candida species present (CFU). Xerostomic drugs and the other variables had no significant effect. CONCLUSION: Older age and more teeth had a significant effect on the number of oral Candida species in the elderly. The results of this study did not support a role of those drugs as a risk indicator for oral Candida carriage. Larger trials are needed to assess the effect of drugs on the presence of oral Candida. PMID- 15819113 TI - Categorisation of dental care provided in the Netherlands. AB - OBJECTIVE: To describe the dental care provided in the Netherlands and investigate to what extent variations can be explained in accordance with certain general characteristics of patients. METHOD: Besides the more usual classification of treatment into groups of dental procedures, in this study it is done using constructed categorisation of dental care based on characteristics such as 'preventive', 'curative', 'new', 'time-consuming', 'complicated' and 'lucrative' treatment. The data were collected from various groups of dentists. Two smaller groups of dentists-informants were asked to categorise dental procedures: 16 out of 55 (29%) and 57 out of 150 (38%). Finally this resulted in six type-scores for each procedure. From another larger group of dentists (n = 607) data were collected about the procedures they performed on a 25% random sample of their patients. Categorisations of dental care per patient were calculated by combining the data on the factual procedures performed with the six 'type-scores' for the procedures concerned. Furthermore, the financial returns dentists generated per patient were calculated from the care they provided. RESULTS: Multilevel analysis shows that considerable variation exists in the categorisations of dental care provided according to age, gender, insurance situation and income level of patients. In youths, for example, relatively more 'preventive' and less 'curative', 'complicated' and 'lucrative' treatment is done and 'older' adults receive relatively less 'preventive' and more 'curative', 'time-consuming' and 'complicated' treatment. CONCLUSIONS: By classifying the dental care provided in certain categories, a general view is obtained of the variations between patients in the care provided. The age of patients appears to be the most critical factor, but there are differences among dentists in the way the age of patients influences their conduct in the provision of dental care. PMID- 15819114 TI - Fluorosis prevalence among schoolchildren in a fluoridated community in Malaysia. AB - OBJECTIVES: This study aims to assess the magnitude of the problem of fluorosis among 10-11 year old schoolchildren in a fluoridated area in Malaysia. METHODS: An analytical cross-sectional design was adopted. Sampling of subjects was by a 2 stage systematic random sampling technique in Selangor, a fully fluoridated area. 1,343 10-11 year olds were assessed for fluorosis using the Dean's index. RESULTS: The prevalence of fluorosis was 58.7% (788 subjects); 478 (35.6%) subjects exhibited very mild fluorosis, 196 (14.6%) mild, 102 (7.6%) moderate, 12 (0.9%) severe and 555 (41.3%) no fluorosis. Tooth prevalence was 30.1%. Overall, the Community Fluorosis Index (CFI) was 0.96 and ranged from 0.23 to 1.72. Fifteen out of 30 schools had CFI > 1.0 (medium public health significance). CONCLUSIONS: The prevalence of dental fluorosis in Malaysia (mostly very mild to mild) at 58.7% is indicative of slightly above optimal levels of exposure. There were pockets of areas where fluorosis were of medium public health significance (CFI > 1.0). It must be cautioned that, fluorosis if not monitored closely, can become an increasing public health concern. PMID- 15819115 TI - Psychometric properties of the modified dental anxiety scale: an independent replication. AB - AIM: To estimate the internal consistency, test-retest reliability, discriminant and concurrent validity of the Modified Dental Anxiety Scale (MDAS). METHOD: Survey of a cohort of 135 dental patients in three equal groups (non-anxious; treated anxious; highly anxious). The average age of the total sample was 39.2 years (SD = 13.21), 101 patients (75%) were female. All patients were asked to complete the MDAS, together with the brief form (six items) of the Speilberger Trait Anxiety Inventory (STAI). Thirty-three participants completed the MDAS on a second occasion 2-3 months after the first testing session. FINDINGS: The MDAS is highly internally consistent (Cronbach alpha = 0.93) and shows good reliability over time (Intra-class correlation coefficient = 0.93). The MDAS was found to discriminate between the three groups of participants defined by self-reported anxiety level, and correlates highly with the Speilberger Trait Anxiety Inventory. CONCLUSIONS: The MDAS is a highly consistent and reliable measure which has good discriminant and concurrent validity. PMID- 15819116 TI - The relationship between children's dental caries experience and the prescription of general anaesthetics. an evaluation of the effectiveness of a Personal Dental Service Pilot. AB - OBJECTIVES: To determine if there was a high correlation between the prescription of a general anaesthetic for dental treatment in young children by a Personal Dental Services (PDS) general anaesthetic pilot and the dental health of children at a ward level. BASIC RESEARCH DESIGN: Postcode data from an audit of the single dental general anaesthetic provider in an area was extracted and mapped to individual wards. A proxy for the oral health population was obtained by using the results of a large-scale dental prevalence study of five-year-olds. The wards were ranked into quintiles by dmft and a mean dmft calculated for each. There were 1,406 patients who had a general anaesthetic, 74% were aged 11 years or less. SETTING: A personal dental service dental general anaesthetic pilot in the north of England providing services for a population of approximately 300,000 residents in a non-fluoridated area. RESULTS There was a positive correlation between increasing levels of dental decay in the population and increasing intervention ratios for dental treatment under general anaesthesia (r2 = 0.90). CONCLUSIONS: The PDS pilot provided dental care under general anaesthesia for more patients from wards with poor dental health than from wards with better dental health. PMID- 15819117 TI - The dental caries experience of 5-year-old children in England and Wales (2003/4) and in Scotland (2002/3). Surveys co-ordinated by the British Association for the Study of Community Dentistry. AB - OBJECTIVE: This paper reports the results of standardized clinical caries examinations of 5 year old children from across England and Wales in 2003/4 and Scotland in 2002/3. These co-ordinated surveys are the latest in a series which seek to monitor the dental health of children and to assess the delivery of dental services. METHOD: The criteria and conventions of the British Association for the Study of Community Dentistry were used. Representative samples were drawn from participating strategic health authorities (SHAs), primary care trusts (PCTs) and health boards (HBs). Caries was diagnosed at the caries into dentine threshold using a visual method without radiography or fibre-optic transillumination. Data for Jersey and the Isle of Man are also included. RESULTS: The results again demonstrated a wide variation in disease prevalence and care strategies across Great Britain. Mean values for d3mft within the current English Strategic Health Authorities ranged from 0.47 in Maidstone Weald (South) to 3.69 in North Kirklees (North); in Wales mean values ranged from 1.48 in Flintshire (NW) to 3.73 in Merthyr (SE); while in Scotland they ranged from 1.29 in Borders to 3.67 in Argyll & Clyde. Mean d3mft across England was 1.49 (d3t = 1.12, mt = 0.19, ft = 0.18), across Wales it was 2.42 (d3t = 1.70, mt = 0.43, ft = 0.29) and across Scotland values were 2.76 (d3t = 1.87, mt = 0.65, ft = 0.24). Overall, 39.6% of children in England & Wales and 55.4% of children inspected in Scotland had evidence of caries experience in dentine (d3mft > 0, including visual dentine caries). The distribution of caries was highly skewed. Thus the mean caries experience for those with dentinal decay in England and Wales was 3.90, as opposed to the overall mean of 1.55; in Scotland the corresponding values were 4.98 and 2.76. Trends over time demonstrate virtually no change in the overall mean d3mft for England and Wales since 2001/2, although the mean value for those with dentine decay experience increased marginally from 3.83 to 3.90. In Scotland there had been deterioration in the overall mean (2.76 as compared to the 2.55 reported in 1999). The care index has also fallen (for example in England & Wales to 12% from 13.2% in 2001/2002 and 14.3% in 1999/2000). CONCLUSION: There has been no overall improvement in the dental health of 5 year old children over the last 2 years. Geographic variation in oral health is marked at both the local and national levels. Overall, the provision of operative care for those with dentinal decay has again decreased slightly. While many children enjoy good oral health, sizable groups remain within the population of 5 year old children who have a clinically significant burden of preventable dental disease. PMID- 15819118 TI - We feel obliged to clarify some of the issues raised in Aubrey Sheiham's editorial. PMID- 15819119 TI - Patient safety. PMID- 15819120 TI - Monitoring the effectiveness of anticoagulation control. AB - PURPOSE: To assess the quality of anticoagulation control at an out-patient clinic and to investigate patterns of deviation from therapeutic ranges. DESIGN/METHODOLOGY/APPROACH: Records for 36,157 clinic visits (2050 patients) were studied. The quality of anticoagulation control was assessed by comparing the measurement of pro-thrombin time recorded at each clinic visit, expressed as an international normalised ratio (INR), with the target therapeutic range for that patient, also recorded at the time of the clinic visit. Each INR measurement was classified according to the relevant patient's therapeutic range and the signed difference between the INR measurement and the centre of the therapeutic range was calculated. For each patient the percentage of their INR measurements that lay within their therapeutic range was calculated. FINDINGS: Of the measurements, 52.3 per cent were within the relevant therapeutic range. The proportion of individual patients' INR measurements within range varied greatly (median 52 per cent, inter-quartile range 40-65 per cent). The quality of anticoagulation control, as measured by the proportion of patients within their therapeutic range, changed little with patient follow-up time. ORIGINALITY/VALUE: The quality of anticoagulation control reported is comparable with that at other centres. The vast majority of patients spend periods outside the therapeutic range for their condition. There may be considerable room for improvement. PMID- 15819121 TI - Managing patients with identical names in the same ward. AB - PURPOSE: To review the experience of managing two patients with identical names in the same ward during a five-month period. DESIGN/METHODOLOGY/APPROACH: The records of the patients were reviewed to look for incorrect entries, errors in specimens sampling, administration of blood products and chemotherapy, and misplacement of clinical notes. Doctors and nurses involved were also invited to complete a questionnaire study to comment on the usefulness of the measures implemented for correct patient identification. A random sample of 60 patients was also selected to see if their full names were shared with other patients attending the same hospital. FINDINGS: Among the 1442 sheets of hospital records from the two patients, no errors pertaining to the clinical activities were found. However, 13 (0.9 per cent) sheets of the hospital records were misplaced. The 21 doctors and nurses participating in the questionnaire study gave positive support to all the additional measures implemented for safeguarding patient identification, of which the automated alerting feature in the electronic clinical management system received the highest scores. A total of 32 (53 per cent) of the 60 sampled patients shared a common full name with one to 101 other patients attending the same hospital. ORIGINALITY/VALUE: Patients with identical names staying in the same ward present a unique challenge to acute health-care settings. The situation is especially relevant in communities where most people's names are not unique. Specific guidelines and measures are needed to prevent patient misidentification. Errors in filing of patient notes and laboratory reports to the hospital record deserve further attention. PMID- 15819122 TI - A volunteer companion-observer intervention reduces falls on an acute aged care ward. AB - PURPOSE: Falls are the most frequently reported adverse event in hospitalised patients and carry a risk of great harm for the frail elderly. This intervention aimed to prevent high-risk in-patients on an acute aged care ward from falling. DESIGN/METHODOLOGY/APPROACH: Patients assessed at high falls risk were accommodated in a room staffed by volunteer companion-observers. The volunteers engaged them in conversation, played cards, opened meals and used the call bell to summon nurses if patients attempted to move from the bed or chair without assistance. Because of occupational health and safety considerations, the volunteers did not assist patients to ambulate. FINDINGS: The falls rate in the acute aged care ward decreased by 44 percent (p < 0.000). No patients fell in the observation room when volunteers were present. Relatives of participating in patients expressed appreciation of the volunteer role, in terms of increased safety and also companionship. Volunteers exercised initiative in determining their pattern of work and developing resources to support their role. RESEARCH LIMITATIONS/IMPLICATIONS: Because volunteers are not present around the clock, other strategies are needed to prevent wandering, frequently confused older in patients from falling during the night. PRACTICAL IMPLICATIONS: In a context where frail elderly patients need constant supervision, using volunteers is a reasonable strategy. ORIGINALITY/VALUE: This intervention used an inexpensive, human resources-based approach to significantly reduce the incidence of falls in the population at highest risk of falling. The additional benefits to patients in terms of cognitive improvement bear further investigation. PMID- 15819123 TI - Quality management and employee commitment: illustrated with examples from Dutch health care. AB - PURPOSE: To assess the relationship between quality management and employee commitment. DESIGN/METHODOLOGY/APPROACH: Analysis of the state of affairs with respect to (TQM) programs in The Netherlands based on the literature and interviews with key informants. FINDINGS: Description of an approach that tries to integrate employee commitment and quality management based on the concept of employees' psychological contracts with their organization (ideas about mutual obligations between them and their employer). RESEARCH LIMITATIONS/IMPLICATIONS: The interviews with key informants provide limited information; the study is done in only one country. Research in a broader context and on a larger scale would give a more general overview. PRACTICAL IMPLICATIONS: The crucial factor in making quality management work can be better described by "quality fails when employees' psychological contracts are neglected" than by "quality fails when system fails". ORIGINALITY/VALUE: This paper highlights the essential relationship between quality management and employee commitment and offers suggestions on how to approach this issue. PMID- 15819124 TI - A generative response to palliative service capacity in Canada. AB - PURPOSE: This paper situates a large-scale learning and service development capacity-building initiative for hospice palliative care services within the current Canadian policy context for use by international readers. DESIGN/METHODOLOGY/APPROACH: In 2000 a national initiative using action research as its design was crafted to support continuing professional development and knowledge management in primary-health care environments. FINDINGS: The Canadian health policy context is complex and requires innovative solutions to achieve desired changes in response to emerging population health demands for quality end of-life care. Employment of educational and social science constructs, including complexity theory, communities of practice, transformative learning theory, and workplace learning methods, has proven helpful in supporting the creation of national capacity for hospice palliative care. RESEARCH LIMITATIONS/IMPLICATIONS: There is a significant contribution for social scientists to make in aiding a better understanding of the complexity in health systems. At the same time, an aging population in industrial countries demands more active engagement of legal and bioethical scholars in a range of emerging policy and legislative questions about quality end-of-life care. Educational research is also required to understand better and reform curricula to prepare an emerging generation of health science practitioners for the demands of an aging population. PRACTICAL IMPLICATIONS: Changing health service delivery environments demand rethinking of the knowledge and skills leaders require to influence desired change. A broader understanding of where and how learning takes place is essential for enhancing the quality of patient care. ORIGINALITY/VALUE: The Pallium Project represents a generative response to facilitating learning and building longer-term system capacity. The journey of project development to date illustrates some important lessons that can be adopted from hospice palliative care to inform other primary health care initiatives, including, potentially, mental health, cardiology, diabetes, geriatrics, where productive change can result from productively linking specialists and primary-care colleagues. PMID- 15819125 TI - Convergence of the health industry. AB - PURPOSE: The purpose of this paper is to define the roles of the key health industry constituents patients, employers, health insurers, hospitals, physicians, Government, pharmaceutical companies and other industry suppliers, plagued with the decline of the health status and affordability in the USA. DESIGN/METHODOLOGY/APPROACH: Reviews literature to back up the author's views. FINDINGS: As health-care demands increase and the constituent roles blur, the industry will experience a gradual convergence. Convergence will redefine and create overlap in the boundaries between the traditional health industry suppliers and in the end this convergence will reshape the future of the health industry. ORIGINALITY/VALUE: predicts that the drivers of change--deteriorating public health, poor quality and safety, and unaffordable health care--will continue to accelerate. Convergence will be gradual and the impact may not be immediately obvious, but the long-term outcome will dramatically reshape the industry. For the major suppliers in the industry, recognizing convergence is only the first step. Navigating convergence so the decisions made by an organization are both productive and profitable, rather than debilitating, is a far more complicated endeavor. The decisions made will not only affect the future of each supplier but also significantly impact the constituent at the center of this industry the patient. PMID- 15819126 TI - The psychological contract. PMID- 15819127 TI - The effects of appreciative inquiry interviews on staff in the U.K. National Health Service. AB - PURPOSE: To obtain preliminary data on the short- and medium-term effects and personal acceptability of appreciative inquiry (AI) in staff development in health care. DESIGN/METHODOLOGY/APPROACH: AI is a non-problem-solving management approach focusing on developing current successes into the future through reflection at individual and group level. Individual one-hour interviews were undertaken with nursing staff on a national paediatric liver in-patient ward. They were asked to recount stories based on their experiences of successful delivery of health care, with active listening, followed by reflection on the process. A total of 32 staff members took part with only two refusals. Data were written and analysed by an open coding method. Follow up was obtained two years later using a written, open question method. FINDINGS: The process was emotional but well received. Staff described quality in interpersonal interactions, preventing errors and engaging their personal values in their work. No improvement in recruitment or retention was shown but a high level of sickness absence fell significantly during the period of the project. Two years later, significant positive effects were recalled and attributed to the interviews by many respondents. AI appears a cost-effective way of connecting professionals' motivation toward quality in their work with strategic intentions. RESEARCH LIMITATIONS/IMPLICATIONS: The interviewer was a medical consultant and ward manager, implying either that the interviews could have worked as a form of managerial supervision or improvements could be a Hawthorne effect. Other unknown influences were likely to be occurring on the ward during the study period. PRACTICAL IMPLICATIONS: Short AI interventions on an individual basis can change sickness absence, at least while the interventions are continuing. It is an important tool for staff motivation with the potential for connecting strategic with micro-operational levels. AI is an approach to NHS management with wide application including appraisal, personal development and mentoring. It can be a positive introduction to reflective practice. ORIGINALITY/VALUE: AI is gaining recognition for its value in staff and service development in health care. The paper shows service and personal effects, cost-effectiveness and illustrates how to use AI for these purposes. PMID- 15819128 TI - International comparisons of patients' views on quality of care. AB - PURPOSE: To compare patients' views on quality of care in different countries using a theory-based instrument, while at the same time controlling for the following potential confounders: type of care system (private vs public), type of care (kind of health problem), gender, age, and subjective wellbeing. DESIGN/METHODOLOGY/APPROACH: Patients capable of communicating in wards (medical and surgical departments) and day surgery departments in England, France, Norway, and Sweden were recruited consecutively, to participate in a programme run by the health-care company Capio. Ward patients: England (n=1236), France (n=1051), Norway (n=226), and Sweden (n=428). Day surgery patients: England (n=887), France (n=544), Norway (n=101), and Sweden (n=742). Average response rate across settings: approximately 75 per cent. Patients evaluated the quality of the care they actually received and the subjective importance they ascribed to different aspects of care. The questionnaire "Quality from the patient's perspective" (QPP) was used (modified short version). FINDINGS: Cross-national comparisons were made within each of the two care contexts (wards and day surgery) separately for men and women. Quality of care evaluations were adjusted for age and subjective wellbeing. English and French patients scored significantly higher than Norwegian and Swedish on both kinds of ratings (perceived reality and subjective importance), in both kinds of care contexts, and in both sexes. ORIGINALITY/VALUE: Cross-national comparisons of patients' views on care can give meaningful guidance for practitioners only if they are context-specific and if well-known confounders are controlled for. PMID- 15819130 TI - [Low dosages of tricyclic antidepressants]. AB - In a meta-analysis of 39 controlled studies, tricyclic antidepressants (TCAs) were found to be effective in the treatment of depressive disorders at dosages under 100 mg/day. No advantage was found for the generally recommended higher dosages. However, the authors ofthe analysis did not pay attention to the problems of diagnostic and therapeutic heterogeneity in the studies that were included. After a closer look, it seems plausible that TCAs do have an effect at lower dosages. Due to the diagnostic heterogeneity, however, it remains unclear whether the effect is on the core symptoms of depressive disorders or rather on symptoms such as anxiety, pain or sleep disorders. Due to the therapeutic heterogeneity, no claims with regard to the effectiveness of lower dosages are justified. The meta-analysis provides no reason to adjust the current guidelines for the dosage of TCAs in the treatment of depressive disorders. PMID- 15819129 TI - [Disorders of concentration and memory in young adults and middle-aged persons]. AB - Four patients, two women aged 29 and 52 and two men aged 46 and 25, respectively, consulted a neurologist for attention and memory disorders. Further investigation revealed that the symptoms were caused by metachromatic leucodystrophy, Graves' disease, Huntington's disease, and a psychological background, respectively. The first patient became dependent in 1.5 years, the second recovered after treatment, the third was independent with slowly progressive symptoms after 1 year, and the fourth was advised to consult a psychologist. Disorders of attention and memory in relatively young people deserve a detailed evaluation at the very first visit, involving the elaboration of an initially extensive differential diagnosis. Too early a separation between a psychic and an organic pathogenesis should be avoided. Indications for the presence of a neurological condition include: consultation at the initiative of others, a relatively brief duration of symptoms without a clear provoking factor, the absence of a psychiatric history or life event, cognitive dysfunction in several areas, abnormal behaviour and an incriminating family history. PMID- 15819131 TI - [Can emerging zoonoses be controlled?]. AB - The report 'Emerging zoonoses' of the Health Council of the Netherlands was written on request from the Minister of Health, Welfare and Sport. It gives an expert view on how to anticipate zoonoses that could emerge in the near future by addressing questions regarding risk assessment, prevention, early detection, control and communication. Further emphasis in the report is given to developments within the European Union (EU). The report had to serve as the basis for a European policy conference on 16 and 17 September 2004 during the Dutch presidency of the EU. Some important developments announced by the Minister are: establishment of a National Centre for Infectious Diseases to advise the Minister and coordinate crisis control during outbreaks, measures to strengthen the regional structure of public health in the Netherlands, and a budget for the strengthening of public health research. PMID- 15819132 TI - [Rheumatoid arthritis: early detection may nip it in the bud]. AB - The finding that IgM-rheumatoid factor and anti-cyclic citrullinated peptide were present in stored blood samples in a significant number of patients with rheumatoid arthritis years before the manifestation of the disease, suggests the disease has a long pre-clinical phase. It might thus be possible to prevent clinical disease by initiating treatment in this pre-clinical phase. Recently a Dutch clinical trial was started with this objective. Patients at risk will be randomised to receive dexamethasone or placebo treatment. PMID- 15819133 TI - [The predictive value of autoantibodies in disseminating lupus erythematosus and rheumatoid arthritis]. AB - In some conditions e.g. osteoporosis, hypertension and hypercholesterolaemia, certain phenomena precede manifestation of the disease and preventive measures can be taken long before the disease presents itself. In the same way systemic lupus erythematosus (SLE) and rheumatoid arthritis can be explored. Recently, studies have been published on healthy blood donors, who later developed SLE or rheumatoid arthritis, and in whom specific autoantibodies could be demonstrated. In SLE the autoantibodies are not specific enough to develop preventive strategies, but in rheumatoid arthritis in particular there are specific antibodies against cyclic citrullinated peptides (anti-CCP-antibodies) which are very specific. A clinical trial has been initiated in which HLA-DR4-positive people with raised autoantibody concentrations are given 1-2 intramuscular injections of dexamethasone with the aim of halving the antibody concentration and in the long term lowering the incidence of rheumatoid arthritis. PMID- 15819134 TI - [Diagnosis of diabetes mellitus: limited use for portable blood-glucose measuring devices]. AB - In the Netherlands, guidelines for the diagnosis of diabetes mellitus are confusing and differ from the international guidelines. Capillary blood-glucose testing using a blood-glucose device is allowed used as a diagnostic tool, although this test is imprecise. The Dutch laboratories measure blood-glucose concentrations by a more precise accurate method, but sometimes measure glucose levels in capillary whole blood and sometimes in venous plasma. These results are not comparable, because the results of capillary measurements are lower than the plasma measurements. In daily practice, health-care professionals are using different methods and are often not aware of the differences in glucose values that may result. They do not realise that glucose devices and laboratory glucose measurements may differ and that capillary- and plasma-glucose values are not interchangeable. Uniformity within the Dutch laboratories with regard to the glucose measurements is urgently needed, as is revision of the Dutch guidelines concerning the diagnosis of diabetes mellitus. This should be based solely on venous plasma-glucose values determined in a laboratory. Portable blood-glucose devices should not be used as a diagnostic tool for diabetes mellitus. These should only be used for blood-glucose control monitoring during treatment or as a screening tool. PMID- 15819135 TI - [Withdrawal symptoms of antidepressants]. AB - Withdrawal symptoms are encountered with both the classical anti-depressants, i.e. the tricyclic antidepressants and monoamineoxidase inhibitors, the modern antidepressants, i.e. the selective serotonin re-uptake inhibitors, and the new antidepressants such as venlafaxine and mirtazapine. The symptoms that are reported following the withdrawal of these drugs can be classified into 8 groups: influenza-like symptoms, psychic symptoms, gastrointestinal symptoms, sleep disorders, equilibrium disorders, sensory disturbances, extrapyramidal symptoms and other symptoms. It is characteristic of these symptoms that they appear 1-4 days after reduction of the dose or the last administration of the drug. They may also appear in the infants of mothers who used an antidepressant during the last phase of the pregnancy. It is important that the patient be informed accurately and carefully at the start and termination of any treatment with an antidepressant. In order to prevent withdrawal symptoms, it is advisable to reduce the dosage slowly if possible when the treatment is to be terminated. PMID- 15819136 TI - [Diagnostic image (233). A girl with nail furrows]. AB - In a 2-year-old girl, transverse furrows of the fingernails were noticed: Beau's lines associated with Kawasaki's syndrome. PMID- 15819137 TI - [Systematic review of the effects of therapy in infants with the KISS-syndrome (kinetic imbalance due to suboccipital strain)]. AB - OBJECTIVE: To establish the effects of manual therapy, chiropractic, or osteopathic treatment of the KISS-syndrome (kinetic imbalance due to suboccipital strain) in infants with positional preference, plagiocephaly, and colic. DESIGN: Systematic review of the literature. METHOD: PubMed, Embase and the Cochrane Library were searched for articles on the effects of manual therapy, chiropractic and osteopathy on the KISS-syndrome. Experts in the field of manual medicine and osteopathy were asked to provide relevant articles. The bibliography in a textbook of manual therapy for children was hand-searched for additional references to the KISS-syndrome. RESULTS: No clinical trials were found that evaluated the effects of manual therapy or osteopathy on either the KISS-syndrome or its symptoms. Pooled analysis of two randomised clinical trials on the effects of chiropractic in infantile colic showed no statistically significant difference between active and control treatments. In addition, we found that 22% of infants showed short episodes of apnoea during manual therapy of the spine, and that one case has been described in which such apnoea resulted in death. CONCLUSION: Given the absence of evidence of beneficial effects of spinal manipulation in infants and in view of its potential risks, manual therapy, chiropractic and osteopathy should not be used in infants with the KISS-syndrome, except within the context of randomised double-blind controlled trials. PMID- 15819138 TI - [Aeromonas caviae infection mimicking inflammatory bowel disease in a child]. AB - A 10-year-old girl presented with a pancolitis with watery bloody diarrhoea. Her family history revealed the presence of inflammatory bowel disease. Whilst awaiting the diagnostic tests results, the stool cultures showed Aeromonas caviae. The disease resolved spontaneously. After 24 months there were no further signs of illness, notably no inflammatory bowel disease. This case represents a more severe form than the mild diarrhoea that is usually described in the literature. In a retrospective search of the hospital database, covering the previous 5 years, 188 patients with positive Aeromonas cultures were found, but the patient described was the only one with bloody diarrhoea with a monoculture positive for A. caviae. PMID- 15819139 TI - [Transitory headaches caused by a colloid cyst of the third ventricle]. AB - A 34-year-old man had a history of short-lasting episodes of rotatory vertigo followed by severe headache, provoked by sudden movements of the head and body. MRI of the brain revealed hydrocephalus secondary to a colloid cyst at the level of the foramen of Monro. The patient underwent microsurgery, after which he remained without symptoms. Colloid cysts are rare, benign tumours accounting for 0.5-1.0% of all primary brain tumours. They are attached by a stalklike appendage to the roof of the third ventricle between the fornices. Typical symptoms include intermittent headache, vomiting, occasional dizziness and blurred vision. These symptoms may be secondary to intermittent obstruction of cerebrospinal-fluid outflow through the foramen of Monro. The results of clinical and neurological examination are usually normal. In any patient with short-lasting episodes of severe headache, provoked by changes in position, an MRI of the brain should be done to exclude a colloid cyst. In general, these patients do not fulfil the criteria of the International Headache Society for migraine because of the short lasting nature of the pain. PMID- 15819140 TI - [Increased risk of stroke during the use of olanzapine or risperidone in patients with dementia]. PMID- 15819141 TI - [Increased risk of stroke during the use of olanzapine or risperidone in patients with dementia]. PMID- 15819142 TI - Building bridges for the scientific advancement of prosthodontics. PMID- 15819143 TI - The profile of patients referred to an outreach restorative dentistry clinic over the last 10 years has changed. AB - The aim of the study was to compare the profile of the patients referred to a Restorative Dentistry consultant clinic at Scarborough General Hospital (SGH) in 1991 and 2001. The clinical records from 80% of a cohort of the new patients referred for consultation and who attended in 1991 and 2001 were examined. In total 55 clinical records were examined from 1991 and 84 from 2001. Data on reason for referral, time of wait before assessment and outcome of the consultation was recorded. The number of patients seen for consultation increased considerably between 1991 and 2001. Mean waiting times from the date of referral to the date when assessed trebled (p = 0.016). Referral for denture problems (notably complete dentures) was the main reason for referral in 2001 (25%). Tooth wear was the most common reason for referral in 1991 (22%). It was concluded that the demand for the services of a Restorative Dentistry consultant at Scarborough General Hospital increased considerably between 1991 and 2001. This resulted in considerably longer waiting times for a consultation. In 2001 there was a greater percentage of referrals for appearance, complete dentures and implants. A higher percentage of patients in this study were referred back to their GDP for treatment as compared to other studies. PMID- 15819144 TI - A study to assess the colour stability of acetal resins subjected to thermocycling. AB - The purpose of this study was to compare colour stability of pink and white acetal resin with one conventional poly (methylmethacrylate) resin. Five specimen disks 50 +/- 0.1 mm in diameter and approximately 1.0 mm thick were prepared from each material. The discs were hydrated for a period of 30 days in distilled water and were then desiccated to a constant weight. Total of fifteen specimens were thermocycled for 100 and 300 hours (4000-12000 cycle) between water baths of 5-55 degrees C. Changes from baseline colour readings were recorded at each interval. Data were analysed with descriptive statistic, repeated measured analysis of variance and single factor analysis of variance. The colour changes for all materials at 100 hours were in the categories of slight, based upon the National Bureau of Standard System (NBS) for expressing colour difference. At 300 hours, the colour changes of materials were significantly different (p = 0.0001). All the materials tested showed clinically acceptable discolouration values. PMID- 15819145 TI - The role of implant surface modifications, shape and material on the success of osseointegrated dental implants. A Cochrane systematic review. AB - We tested the hypothesis of no difference in implant failures between various dental implant types. We searched for all randomised clinical trials comparing different implant types/systems with a follow up of at least one year on four databases. Screening of eligible trials, quality assessment and data extraction were conducted in duplicate. Thirty-one trials were identified. Twelve trials, reporting results of 512 patients, were included. No significant differences were observed for implant failures. There were minor statistically significant differences for peri-implant bone level changes. Turned surfaces had a 20% reduction in risk of being affected by perimplantitis over a 3-year period. PMID- 15819146 TI - Retention of quartz-fibre endodontic posts with a self-adhesive dual cure resin cement. AB - This study examines retention of a quartz-fibre endodontic post cemented in extracted human teeth using two cement types. Fifty-four extracted buman central incisor teeth were prepared for a tapered quartz-fibre post. Posts were cemented in 28 teeth using conventional resin cement (RelyX ARC), and 26 teeth using self adhesive resin cement (RelyX Unicem). Posts were cyclically-loaded. Bond strengths were tested and dependability assessment was carried out. Although there was no significant difference in Weibull modulus between groups (p < 0.05), greater dependability was displayed by RelyX Unicem group. Easy application and greater dependability of self-adhesive cement may improve bonding of fibre-based posts in root canals. PMID- 15819147 TI - Retrospective study on the performance of porcelain laminate veneers delivered by undergraduate dental students. AB - The aim of this study was to retrospectively evaluate the clinical performance of porcelain laminate veneer restorations delivered by undergraduate students at the Department of Restorative Dentistry, University Dental School and Hospital Cork and to analyze the factors that influence their prognosis and their effect on periodontal health. Patients who had veneers fitted over a period of 5 yrs. were recalled for evaluation. The recall protocol took the form of a questionnaire and a full clinical examination of the teeth involved and contra lateral control teeth. In all, 29 patients with 62 porcelain veneers were examined, only 2 of which were in the mandible. 77% of patients had veneers fitted due to discolouration, 13% in order to alter the shape or size of teeth and 10% due to palatal erosion. On the date of recall 89% of veneers had never debonded or fractured, 6% had to be rebonded since the date of insertion and 5% presented with a fracture. From this study it appears that porcelain laminate veneers are successful in the treatment of discoloured and irregular configured anterior teeth. It shows that undergraduate students can deliver satisfactory veneer restorations. PMID- 15819148 TI - Instant orthodontics? PMID- 15819149 TI - Why does local anaesthesia not work everytime? AB - This paper describes the reasons why local anaesthesia may fail. It offers strategies to help overcome failure of local anaesthesia in dentistry. PMID- 15819150 TI - A review of gagging problems in dentistry: 2. Clinical assessment and management. AB - A pronounced gag reflex can be a severe limitation to a patient's ability to accept dental care and for a clinician's ability to provide it. It can compromise all aspects of dentistry from diagnostic procedures to active treatment and can be distressing for all concerned. Many 'management' techniques have been described. This paper describes the different categories of treatment used to manage people with pronounced gag reflexes. PMID- 15819151 TI - Management of inhaled and swallowed foreign bodies. AB - Although accidental inhalation or ingestion episodes are very rare, the consequences can be very serious for the patient, so it is important when the total number of courses of dental treatment performed each year are considered. This article discusses the causes, likely sequelae and management strategies adopted following such incidents. PMID- 15819152 TI - Relaxation and complementary therapies: an alternative approach to managing dental anxiety in clinical practice. AB - Dental anxiety is a widespread problem and has a significant impact on the provision of dental care for the general population. Anxiety leads to avoidance of dental treatment and increased stress for dental practitioners. Traditionally, dental anxiety has been managed using pharmacological techniques. This article reviews alternative treatment approaches, including psychological and complementary therapies which can be used in managing dental anxiety and facilitating dental treatment. Many of these approaches are currently being used within the dental profession. Others are gaining a wider acceptance as to their usage. Where available, the evidence of their efficacy will be reviewed. PMID- 15819153 TI - Burkitt's lymphoma: an unusual cause of lip numbness. AB - A 38-year-old man was referred by his dental surgeon with a painful swelling in the right mandibular sulcus and an ipsilateral numb lip. Dental infection secondary to caries was initially diagnosed but biopsy revealed this to be a Burkitt's lymphoma. After intense chemotherapy, the patient is still in remission three years later. This case is a reminder that malignancy should always be high on a clinician's list of differential diagnoses of jaw swelling associated with neurological abnormalities and prompt referral is essential. PMID- 15819154 TI - An overview of bleaching techniques: 3. In-surgery or power bleaching. AB - The use of in-surgery bleaching techniques has increased recently within the profession following the introduction of various new systems. Many dentists and patients alike prefer the in-surgery procedures to lighten teeth as it takes the responsibility for the procedure firmly away from the patient to the dentist; a situation with which both sides may be more comfortable. This article will review various in-surgery or power bleaching procedures. In addition, it will detail the procedure involved and how and when to combine bleaching techniques. PMID- 15819155 TI - Radicular cyst arising from a traumatized primary incisor: a case report of a rare complication that emphasizes the need for regular follow up. AB - Radicular cysts associated with the deciduous dentition are rare. The literature is sparse regarding cysts associated with primary incisor teeth, which is surprising given the frequency with which these teeth are traumatized. Most cases reported involve molar teeth associated with apical infection following on from caries. A case of a 3-year-old boy who developed a large radicular cyst associated with a previously traumatized upper primary incisor is reported. PMID- 15819156 TI - Restorative materials. PMID- 15819157 TI - Oral cancer risk in diabetic patients. PMID- 15819160 TI - Last-minute survey preparation can pay off with big dividends. PMID- 15819159 TI - Physical signs for the general dental practitioner. PMID- 15819161 TI - Data processes: JCAHO's 'data-driven' agenda. PMID- 15819162 TI - Discharge planning advisor. Patients placed quickly with software combo. PMID- 15819163 TI - Evaluating QI efforts through collection tools. PMID- 15819164 TI - Performance measures depend on effective use of good data. PMID- 15819165 TI - Patient safety alert. Study targets errors in ambulatory setting. PMID- 15819166 TI - Fecal leukocyte proteins in inflammatory bowel disease and irritable bowel syndrome. AB - The aim of this prospective study was to compare five different leukocyte proteins in feces of patients with chronic inflammatory bowel disease (IBD), irritable bowel syndrome (IBS) and healthy persons who underwent prophylactic colonoscopy. METHODS: The leukocyte proteins calprotectin, lactoferrin, lysozyme, myeloperoxidase, and PMN-elastase were determined with immunoassays in fecal samples of three consecutive feces (e.g. three days) in 40 healthy persons, 39 patients with chronic IBD (of these 21 with Crohn's disease and 18 with ulcerative colitis), and 40 patients with IBS. RESULTS: ROC curves calculated for healthy persons and patients with IBD yielded the following areas under the curves (AUCs): PMN-elastase 0.916, calprotectin 0.872, myeloperoxidase 0.750, lysozyme 0.726, and lactoferrin 0.693. The AUCs of PMN-elastase and calprotectin were not significantly different (p = 0.327), whereas PMN-elastase or calprotectin vs. the other proteins were significantly different (p < 0.001). PMN elastase and calprotectin correlated with the endoscopically classified severity of inflammation. All fecal leukocyte markers in IBS were found in the range of the healthy persons. Data on storage stability of leukocyte proteins in fecal supernatants are given. CONCLUSION: Fecal PMN-elastase and calprotectin support the differentiation of chronic IBD from IBS and correlate with the severity of inflammation. PMID- 15819167 TI - Clinical use of serum prostate-specific antigen: a review. AB - Serum Prostate-Specific Antigen (PSA), in conjunction with digital rectal examination, is a common first-line clinical tool for the diagnosis of prostate cancer; therefore, it is essential that the clinical use of PSA be further developed to make it a more definitive tool. There are issues in screening with PSA, with regard to its lack of specificity (and sensitivity) and solutions involving various PSA parameters and derivatives have been suggested to address these issues. The literature on these parameters and research into factors found to influence levels of PSA are discussed in this review. PMID- 15819168 TI - Dysgammaglobulinaemia in the elderly--a review and case studies. AB - An understanding of the possible causes of dysgammaglobulinaemia in the elderly helps to direct further investigation to establish a diagnosis. In this review we provide brief case studies to illustrate some of the disorders associated with dysgammaglobulinaemia in the elderly. We consider both hypergammaglobulinaemia (polyclonal, characteristic of chronic inflammatory disorders or autoimmunity, and monoclonal, often with an associated malignant disorder) and hypogammaglobulinaemia (including immunodeficiency, immune paresis secondary to malignancy and protein loss). Where dysgammaglobulinaemia is noted in the elderly the most useful laboratory tools to help discern the pathogenesis are serum and urine electrophoresis, autoantibody investigations and measurement of liver and renal function. PMID- 15819169 TI - Stability of insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-3 measured by the IMMULITE automated chemiluminescence assay system in different blood specimens. AB - Insulin-like growth factor I (IGF-I) and IGF binding protein 3 (IGFBP-3) are measured to diagnose disorders of the somatotropic axis in children and adults. In clinical studies samples for IGF-I and IGFBP-3 measurement must often be stored for months and sent to specialized laboratories. Therefore, we tested the stability of IGF-I and IGFBP-3 in whole blood, serum and plasma from 12 volunteers at 4 degrees, 22 degrees, and 37 degrees C for several hours and at 25 degrees C for several months. The effect of only one protease inhibitor (Aprotinin = Trasylol, Bayer, Germany ) on IGF-I and IGFBP-3 measured in the automated IMMULITE assay system (DPC, Los Angeles) was tested. IGF-I and IGFBP-3 were stable in heparinized whole blood, plasma and serum at 22 degrees C up to 24 hours. IGF-I was stabilized by aprotinin for up to 72 hours at 37 degrees C. Factor concentrations were not altered after storage at -25 degrees C for at least 12 months. Recognition of IGFBP-3 fragments by the antibody used in the automated IGFBP-3 IMMULITE was excluded by measurement in 26 sera from pregnant women which usually contain IGFBP-3 fragments. In conclusion, samples for measurement of IGF-I and IGFBP-3 should be kept on ice and cooled if shipment takes more than 48 hours or alternatively 5000 IU/ml aprotinin should be added. IMMULITE assays are also valid to measure IGF-I and IGFBP-3 after at least 12 months storage at -25 degrees C. PMID- 15819170 TI - "Method-specific" stability of serum C-peptide in a multicenter clinical study. AB - A Central laboratory that participates in multicenter clinical studies must consider and study the parameters that condition the in vitro stability of the analytes. We evaluated the effect of temperature on serum C-peptide storage during transport from clinical research centers to the Central laboratory. In particular, the stability for storage lengths from 0 to 24-48 hours at temperatures of between -20 degrees C and +37 degrees C were studied: the C peptide assay was performed by means of a chemiluminescence and a RIA method. The tests confirmed that sample freezing is the gold standard for accurate determination of serum C-peptide and that storage at 37 degrees C may decrease the analyte levels. Instead, C-peptide stability at 2-8 degrees C appeared "method-specific"; while no apparent alteration was obtained with the chemiluminescence method up to 24 hours of storage, the RIA showed an early slight increase in C-peptide that is proportionate to storage time. Our work highlights that before starting up a multicenter clinical study it is always necessary to optimize and standardize biological sample storage and transport conditions to guarantee a high quality sample for analysis. Beyond this, it is even very useful to check the reliability of technical and instrumental resources that the Central laboratory will use during the study because molecular alterations of the analytes due to variable storage conditions can cause misleading results. PMID- 15819171 TI - Effect of heparins on thrombin generation in hemophilic plasma supplemented with FVIII, FVIIa, or FEIBA. AB - Central venous catheters assist infusion of coagulation factors in hemophiliacs but can be problematic due to mechanical dysfunction, infection, and thrombosis. The effect of low molecular weight heparin, unfractionated heparin, or covalent antithrombin-heparin complex on thrombin generation in factor concentrate supplemented hemophilic plasma were studied. Thrombin levels were similar to normal plasma after the addition of factor eight inhibitor bypassing agent to hemophilic plasma. At 0.2 U/ml of heparin, covalent antithrombin-heparin inhibited free thrombin generation to a greater degree than heparin and low molecular weight heparin. Covalent anti-thrombin-heparin may give a greater anticoagulant response in hemophilic plasma supplemented with factor VIII or factor VIIa than with factor eight inhibitor bypassing agent. Requirements for heparin in hemophilic patients with thrombosis may depend on the procoagulant treatment used. PMID- 15819172 TI - N-terminal pro-brain natriuretic peptide (NT-proBNP) in healthy blood donors and in patients from general practitioners with and without a diagnosis of cardiac disease. AB - BACKGROUND: The natriuretic peptides and especially the N-terminal pro-brain natriuretic peptide (NT-proBNP) are increased in conditions with cardiac ventricular volume and pressure overload. Early stages of ventricular volume and pressure overload are often without signs and symptoms and therefore difficult to diagnose. On the contrary, a normal level of a natriuretic peptide excludes congestive heart failure as a cause of dyspnea with high probability. In addition, natriuretic peptide levels predict the risk of death and cardiovascular events after adjustment for traditional risk factors. A few studies suggest that age, gender and renal function may influence circulating natriuretic peptide levels. This study was therefore initiated to a) assess reference values for the N-terminal pro-brain natriuretic peptide (NT-proBNP) in a group of blood donors and healthy elderly individuals and to relate these levels to age, sex, and creatinine and b) to measure the levels of NT-proBNP in a population of patients presenting to general practitioners and to check the quality of the diagnoses congestive heart failure and dyspnea of other causes (heart failure patients usually present with breathlessness but the low specificity of dyspnea often leads to misdiagnoses). Finally, the percentage of patients with other diagnoses and elevated NT-proBNP as an indicator of an increased cardiovascular risk or up to now unknown cardiac disease was determined. STUDY DESIGN AND METHODS: N=1981 blood donors, N=283 individuals from general practitioners (GP) without cardiac disease and N=570 consecutive patients from GPs were recruited and tested for the presence of NT-proBNP using a newly developed electrochemiluminescence immunoassay run on an automated analyzer (Elecsys, Roche Diagnostics). RESULTS: NT-proBNP was detected at relatively homogenous levels in all individuals below the age of 50 years. NT-proBNP values increased with increasing age which was due to the increasing number of outliers in that group. Females had higher NT-proBNP levels than males. CONCLUSIONS: Based on the assumption that individuals below the age of 50 years are healthy, reference values based on the 97.5th percentile were established. These values were considered to be normal. The presented data and data from the literature suggest that also in the elderly population a cut off level of 125 pg/ml is useful either to exclude cardiac dysfunction in symptomatic individuals or to risk stratify elderly individuals in terms of the necessity for intervention. PMID- 15819173 TI - Thrombosis and bleeding: when opposites are not so far apart. AB - Thrombotic events in patients with inherited bleeding disorders occur only rarely. However, in some cases, the co-existence of acquired or inherited prothrombotic risk factors may overcome the hypocoagulative state, modulating the clinical phenotype to a decrease in bleeding symptoms or even to an increase in the likelihood of developing thrombotic complications. This review summarizes the cases of thrombosis reported in the literature and analyzes the most important risk factors for thrombosis in patients with a congenital bleeding tendency. Data were identified by searches of the published literature, including PubMed, references from reviews and abstracts from the most important meetings on this topic. There is increasing evidence that thrombotic complications in patients with hereditary bleeding disorders have a multifactorial pathogenesis, depending on acquired (coagulation factor replacement therapy, central venous catheters, surgery, viral infections) and/or inherited (thrombophilic gene mutations) prothrombotic risk factors. PMID- 15819174 TI - Viral diarrhea in Japanese children: results from a one-year epidemiologic study. AB - A total of 557 fecal specimens from infants and children with acute gastroenteritis in five places (Maizuru, Tokyo, Sapporo, Saga and Osaka) in Japan from July 2002 to June 2003 were tested for the presence of diarrheal viruses by RT-PCR, PRHA, RNA-PAGE and latex agglutination methods. Of these, 56.4% (314) were found positive for diarrheal viruses. Among them, group A rotavirus was the most prevalent (43.6%, 137 of 314) followed by norovirus (29.9%, 94 of 314), adenovirus (7.6%, 24 of 314), group C rotavirus (6.4%, 20 of 314), sapovirus (5.1%, 16 of 314) and astrovirus (1.6%, 5 of 314), respectively. A high rate (7.4%, 19 of 314) of viral mixed infections, including one triple infection (adenovirus, norovirus and astrovirus) was demonstrated. Norovirus infection that usually has a peak during November and January in Japan was detected year-round and highest in September in our study. Norovirus was subjected to molecular genetic analysis by sequencing. The results clearly indicated that norovirus group II was a dominant genogroup (94.3%, 100 of 106). It is noteworthy that noroviruses detected in this study were classified into 8 genotypes (GI/1, GI/4, GII/2, GII/3, GII/4, GII/5, GII/6 and GII/12). Of these, NVGII/4 was the predominant genotype, followed by NVGII/6, and these presented 75.6% (80 of 106) and 11.3% (12 of 106), respectively. Another interesting feature in our study was the sudden appearance and disappearance of SaitamaU16-like strains belonging to NVGII/6 in the short period (January 2003 to June 2003). Our findings confirmed the presence of many diarrheal viruses co-circulating among Japanese infants and children and showed the great genetic diversity among norovirus. PMID- 15819175 TI - The latest from the IVD industry December 2004. Chlamydia--pathogens that are still often underestimated. AB - Chlamydiae are bacteria living as intracellular parasites. Contemporary differentiation distinguishes nine species, of which three are relevant to human medicine. Infections with Chlamydia trachomatis are among the most widespread sexually transmitted diseases. Chlamydophila pneumoniae invade the respiratory tract, chlamydophila psittaci is the pathogen of ornithosis. Chlamydial infections frequently progress asymptomatically and hence remain untreated. Serious chronic secondary diseases are the outcome; they are protracted and cost intensive in treatment. Besides the detection of the pathogen, serology plays a vital part in diagnostics. The antibiotic therapy of chlamydial infections involves the application of tetracyclines, macrolides or chinolons. Chlamydophila pneumoniae in particular has become focus of medical interest as one of the causes of arteriosclerosis. PMID- 15819176 TI - The latest from the IVD industry. December 2004. Campylobacter widespread among wild and domestic animals. AB - Infection with campylobacter is a worldwide anthropozoonosis and considered to be the most frequent frequent bacterial cause of infectious enteritis [1]. The disease takes a variety of courses and in past years has actually overtaken the number of salmonellal infections in some industrialized countries. PMID- 15819177 TI - [Fall 2004 Congress of the German Society of Laboratory Physicians]. PMID- 15819178 TI - Promoting health in children and young people: identifying priorities. PMID- 15819179 TI - Breastfeeding for longer: what works? PMID- 15819180 TI - Promoting the social and emotional health of children: where to now? AB - In advanced industrial societies such as the UK, the burden of disease is shifting from physical to mental health problems--emotional and behavioural problems currently being the major cause of disability in children. Evidence concerning the role of parents in promoting children's social and emotional health, in conjunction with the benefits of supporting parents in this role, has focused attention on the need for greater clarity concerning the role of both parents and governments in supporting and protecting children's social and emotional health. This paper examines the evidence base concerning children's social and emotional development and suggests some steps that may be necessary to ensure the future promotion of children's social and emotional health. PMID- 15819181 TI - Supporting young people who repeatedly self-harm. AB - This paper explores the views and experiences of a group of 74 young people aged 16-22 who were interviewed following presentation in accident and emergency (A&E) departments after intentionally harming themselves. It focuses on a sub-group of 38 young people with a history of self-harm behaviour that extended from when they were under the age of 16, one-third of whom had been or were currently in care. Whilst some had kept their self-harming hidden and had not received any professional intervention until they reached adulthood, others had been in touch with services, although their treatment had not prevented them from continuing to self-harm. Medication was perceived as 'fobbing off', particularly when unaccompanied by other treatments. The young people described their encounters with counsellors and clinicians, some of whom they perceived not to understand or to listen to their perspective. Whilst not representative of all young people who self-harm, these views are important and deserve attention if interventions for children and adolescents are to prevent repetition of self-harm. PMID- 15819182 TI - A review of low birth weight: predictors, precursors and morbidity outcomes. AB - This paper reviews the global evidence concerning infant low birth weight (LBW) (2.500kg and below) in relation to suggested causative factors, such as undernutrition, stress, smoking, drug abuse and deprived living environments. In addition, health promotion strategies in place to reduce LBW incidence in both developed and underdeveloped countries are reviewed. The paper also focuses on the high incidence of LBW in the UK in comparison to other European and developed countries. Predictors of LBW are then considered comparing findings of a study conducted in the UK with findings in underdeveloped countries. In addition, LBW risk is discussed in the context of vulnerability factors, such as teen pregnancy, child abuse and domestic violence. In conclusion, the author argues for a health promotion policy in the UK to reduce incidence of LBW in areas of deprivation and suggests that the improvement of maternal health is key in improving the health of today's infants and children who will one day become the adults of tomorrow. PMID- 15819183 TI - Risks and benefits of the single versus the triple MMR vaccine: how can health professionals reassure parents? AB - Measles, mumps and rubella (MMR) are all preventable but infectious diseases caused by viruses. A particular study by Wakefield et al suggests that there are potentially adverse effects of having the triple MMR vaccine. This has been reported widely by the media and has caused alarm to parents of young children, probably contributing to the decline in its uptake. In order to provide the context for the debate regarding the single versus the triple vaccine, this paper briefly appraises firstly, the Wakefield et al research paper that has led to public health concerns and secondly, a more rigorous research study (Madsen et al) that contradicts the findings; the paper then explores the risks and benefits of the single and the triple MMR vaccine programmes, finally providing a short discussion on factors that might influence the decision-making process by parents when faced with the dilemma of not having their child vaccinated, or opting for either the single or triple vaccination programme. PMID- 15819184 TI - Anxiety, depression and smoking in schoolchildren--implications for smoking prevention. AB - The prevalence of smoking among 14 to 18-year-old adolescents (one out of three) is as high as in the general population in Hungary. Depression and anxiety disorders are also major public health problems. Our objective was to investigate the correlations between smoking status and anxiety and depressive symptoms, as well as to identify potential implications for the prevention of nicotine dependence. We used a self-developed questionnaire to survey smoking behaviour, and standard self-evaluation tests (the Hungarian version of the Spielberger's State-Trait Anxiety Inventory for Children and the Children's Depression Inventory) to capture anxiety and depressive symptoms in 215 students from randomly selected technical and high schools. Smoking students showed significantly more anxiety and depressive symptoms compared to their non-smoking peers. Importantly, occasional smokers appeared to be more anxious than non smokers, whereas daily smokers scored higher on the depression inventory than either non-smokers or occasional smokers. The majority of students with high scores suggestive of anxiety or depressive disorders were smokers. Interestingly, Levels of anxiety and depression were associated with smoking status in boys but not in girls. The strong association found between anxiety and depressive symptoms and youth smoking, particularly in boys, suggests that reducing these symptoms (especially high, school-related anxiety) may be preventive against the onset of addictive smoking. This, together with the high level of anxiety and depression found in girls, makes it essential for all professionals dealing with adolescents and education to recognise these symptoms and their significance, and to be aware of the ways of reducing them by changes in the educational system, or medically, if necessary. PMID- 15819185 TI - Fetal gender determination and the male:female population ratio in Bangladesh. AB - Gender determination of the fetus can be an important issue to parents, especially in Bangladesh and other South Asian countries. This study was conducted to determine how often we can detect the gender of the fetus and whether the gender ratio of the fetus correlated with the gender ratio of the population as determined by the census. Fetal gender ratio of male and female was found to be 1.04:1; the population gender ratio of the same period was 1.038:1. This study was conducted from March 1999 to December 2001. PMID- 15819186 TI - Atmospheric processes may create perchlorate. PMID- 15819187 TI - Common arsenical pesticide under scrutiny. PMID- 15819188 TI - Proposed selenium standard under attack. PMID- 15819189 TI - Allocating cleanup costs at hazardous waste sites. PMID- 15819190 TI - On-line analysis of gas-phase composition in the combustion chamber and particle emission characteristics during combustion of wood and waste in a small batch reactor. AB - The emission of particulate matter and gaseous compounds during combustion of wood and refuse-derived fuel in a small batch reactor is investigated by laser mass-spectrometric on-line measurement techniques for gas-phase analysis and simultaneous registration of physical aerosol properties (number size distribution). The gas-phase composition is addressed by a laser-based mass spectrometric method, namely, vacuum-UV single-photon ionization time-of-flight mass spectrometry (VUV-SPI-TOFMS). Particle-size distributions are measured with a scanning mobility particle sizer. Furthermore, a photoelectric aerosol sensor is applied for detection of particle-bound polycyclic aromatic hydrocarbons. The different phases of wood combustion are distinguishable by both the chemical profiles of gas-phase components (e.g., polycyclic aromatic hydrocarbons, PAH) and the particle-size distribution. Furthermore, short disturbances of the combustion process due to air supply shortages are investigated regarding their effect on particle-size distribution and gas-phase composition, respectively. It is shown that the combustion conditions strongly influence the particle-size distribution as well as on the emission of particle-bound polycyclic aromatic hydrocarbons. PMID- 15819191 TI - Temporal changes in U.S. benzene emissions inferred from atmospheric measurements. AB - The 1990 Clean Air Act Amendments required the United States Environmental Protection Agency (U.S. EPA) to enact stricter regulations aimed at reducing benzene emissions. In an effort to determine whether these new regulations have been successful in reducing atmospheric benzene concentrations, we have evaluated benzene-to-acetylene ratios from data sets spanning nearlythree decades, collected during several field studies and from the U.S. EPA's Photochemical Assessment Monitoring Station (PAMS) network. The field-study data indicate a decrease in benzene relative to acetylene of approximately 40% from 1994 to 2002. This corresponds to a decrease in benzene alone of approximately 56% over the same period. In contrast, the PAMS data exhibit high interannual variability with no discernible trend. This discrepancy is attributed to measurement problems in the PAMS data sets. PMID- 15819192 TI - Single-particle characterization of four "Asian Dust" samples collected in Korea, using low-Z particle electron probe X-ray microanalysis. AB - A single-particle analytical technique, named low-Z particle electron probe X-ray microanalysis (low-Z particle EPMA), employing an ultrathin window X-ray detector and enabling the quantitative determination of even low-Z elements such as C, N, and O, is applied to characterize "Asian Dust" samples, collected in ChunCheon, Korea, during four Asian Dust storm events on March 7, 2000, April 7, 2000, March 22, 2001, and May 17, 2001. In this study, it is demonstrated that single particle analysis using the low-Z particle EPMA provides detailed information on various types of chemical species in the samples. The most abundantly encountered particles, both in coarse and fine fractions, are aluminosilicates. The relative abundances of those particles on the basis of their size are different between the four Asian Dust samples. The sample collected on March 7, 2000 did not experience any chemical modification during its transport because the sample does not contain particles of chemical species that result from atmospheric reactions. The sample collected on April 7, 2000 contains both genuine and reacted sea-salt particles. The genuine sea-salts are in the form of a mixture of NaCl and MgCl2 entrained during their passage over the Yellow Sea. The reacted sea-salts particles are encountered very much in fine fraction. The sample collected on March 22, 2001 shows somewhat significant chemical modification both for CaCO3 and sea-salts particles. For this sample, a significant number of reacted CaCO3 and sea-salt particles, such as those containing nitrate and/or sulfate, are encountered, implying that CaCO3 and sea-salts particles have reacted with sulfur or nitrogen oxide species during their long-range transport. The sample collected on May 17, 2001 experienced the most extensive chemical modification during its transport. In addition to the observation of the extensively reacted CaCO3 and sea-salt particles, reacted K2CO3 particles are also extensively encountered in this sample, which implies that K2CO3 species should be regarded as an additional important chemical species in the study of the chemical modification of Asian Dust particles during long-range transport. PMID- 15819193 TI - Co-occurrence of triclocarban and triclosan in U.S. water resources. AB - Triclocarban (TCC) and triclosan (TCS) are antimicrobial additives in personal care products. Whereas TCS has been studied extensively, the environmental fate of TCC remains largely unknown. To address this data gap, we performed quantitative structure-activity relationship (QSAR) analyses that suggested a propensity of TCC to persist in various environmental compartments with predicted half-lives ranging from 0.75 days in air to 540 days in sediment. Moreover, concentrations of both antimicrobials were measured in 42 environmental samples from the Greater Baltimore region using a combination of solid-phase extraction, liquid chromatography/mass spectrometry, and isotope dilution. The co-occurrence of TCC and TCS was observed, owing to similar properties, usage, disposal, and environmental half-lives. A linear empirical correlation (R2 = 0.9882) fit the log-log-transformed data from diverse aquatic media and spanned 5 orders of magnitude in concentration. Occurrences of TCC predicted for 85 U.S. streams were statistically indistinguishable from experimental regional data (alpha < or = 0.05). Annual loading of antimicrobials to water resources probably is dominated by activated sludge treatment plants (39-67%), followed by trickling filters (31 54%) and combined and sanitary sewer overflows (2-7% and <0.2%, respectively). Study results suggest that TCC is a previously unrecognized contaminant of U.S. water resources nationwide, likely ranking in the top 10 in occurrence rate and in the top 20 in maximum concentration among 96 organic pollutants considered. The magnitude and frequency of TCC contamination (regional, 6750 ng/L, 68%; predicted nationwide for 1999--2000, 1150 ng/L, 60%) were markedly higher than non-peer-reviewed numbers (240 ng/L, 30%, U.S.) currently used by the U.S. Environmental Protection Agency for evaluating TCC's ecological and human health risks. PMID- 15819194 TI - Estrogenic activity in sediments from European mountain lakes. AB - Superficial and bottom sediment samples from 83 European mountain lakes, ranging from Norway to the Pyrenees and East Europe, were tested for estrogenic compounds by the recombinant yeast assay. The results showed widespread potential estrogenic activity arriving at remote lakes. Tatra Mountains (Slovakia) and Scotland Highlands were the regions with the highest prevalence of lakes with high estrogenic values. Comparison of the estrogenic activity in the superficial layer of sediments with pre-industrial age sections showed that estrogenic compounds were predominantly deposited in recent times. Chemical analysis showed that highly estrogenic sediments were significantly enriched in both polycyclic aromatic hydrocarbons (PAH) and organochlorine compounds. For PAH, enrichment ratios in highly estrogenic samples versus nonestrogenic ones were inversely correlated with the vapor pressure value for each compound, indicating a significant relationship between estrogenicity and accumulation of less volatile PAH. Two PAH of predominantly diagenetic origin, retene and perylene, did not show specific enrichment in estrogenic samples. Principal component analysis revealed a strong correlation between estrogenic activity and the presence of contaminants of anthropogenic origin. These data reveal significant amounts of estrogenic compounds in remote lakes, relate them to the overall human activity, and suggest that they may affect organisms inhabiting these ecosystems. PMID- 15819195 TI - Emission factors and importance of PCDD/Fs, PCBs, PCNs, PAHs and PM10 from the domestic burning of coal and wood in the U.K. AB - This paper presents emission factors (EFs) derived for a range of persistent organic pollutants (POPs) when coal and wood were subject to controlled burning experiments, designed to simulate domestic burning for space heating. A wide range of POPs were emitted, with emissions from coal being higher than those from wood. Highest EFs were obtained for particulate matter, PM10, (approximately 10 g/kg fuel) and polycyclic aromatic hydrocarbons (approximately 100 mg/ kg fuel for sigmaPAHs). For chlorinated compounds, EFs were highest for polychlorinated biphenyls (PCBs), with polychlorinated naphthalenes (PCNs), dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) being less abundant. EFs were on the order of 1000 ng/kg fuel for sigmaPCBs, 100s ng/ kg fuel for sigmaPCNs and 100 ng/kg fuel for sigmaPCDD/Fs. The study confirmed that mono- to trichlorinated dibenzofurans, Cl1,2,3DFs, were strong indicators of low temperature combustion processes, such as the domestic burning of coal and wood. It is concluded that numerous PCB and PCN congeners are routinely formed during the combustion of solid fuels. However, their combined emissions from the domestic burning of coal and wood would contribute only a few percent to annual U.K. emission estimates. Emissions of PAHs and PM10 were major contributors to U.K. national emission inventories. Major emissions were found from the domestic burning for Cl1,2,3DFs, while the contribution of PCDD/F-sigmaTEQ to total U.K. emissions was minor. PMID- 15819196 TI - Comparison of toxaphene congeners levels in five seal species from eastern Canada: what is the importance of biological factors? AB - Environmentally relevant chlorobornanes (CHBs) were measured in blubber samples of harbor (Phoca vitulina), gray (Halichoerus grypus), harp (Phoca groenlandica), and hooded seals (Cystophora cristata) sampled in different part of the St. Lawrence marine ecosystem (SLME) and ringed seals (Phoca hispida) sampled in the eastern Canadian Arctic waters. The purpose of this study was to compare the levels of six CHBs (Parlar-26, -40/-41, -44, -50, and -62) among the five seal species. Seal species could be separated into three groups based on their respective sigmaCHB mean concentrations (+/-standard error): gray (49+/-3.9 ng/g lipid weight) and harbor (80+/-20 ng/g lipid weight) seals were more contaminated than ringed seals (18+/-7.6 ng/g lipid weight) but less contaminated than harp (370+/-87 ng/g lipid weight) and hooded (680+/-310 ng/g lipid weight) seals. These differences are not expected to be related to different sources of toxaphene contamination, since both the SLME and the eastern Canadian Arctic environments are thought to be mainly contaminated via atmospheric transportfrom the southeastern part of the United States. Thus, biological factors such as sex, age, nutritive condition, metabolism capacity, and diet of the animals collected were considered. Results reported in this study indicated that the diet is likely the main factor accounting for interspecies variations in toxaphene contamination in seals from eastern Canada. PMID- 15819197 TI - Freshwater mussel shells as environmental chronicles: geochemical and taphonomic signatures of mercury-related extirpations in the North Fork Holston River, Virginia. AB - This study utilized freshwater mussel shells to assess mercury (Hg) contamination in the North Fork Holston River that extirpated (caused local extinctions of) a diverse mussel fauna. Shells (n = 366) were collected from five sites situated upstream (two sites), just below (one site), and downstream (two sites) of the town of Saltville, Virginia, where Hg was used to produce chlorine and caustic soda from 1950 to 1972. Shell samples were used to test the (1) utility of geochemical signatures of shells for assessing the spatial variation in Hg levels in the river relative to the contamination source and (2) value of taphonomy (post-mortem shell alteration) for distinguishing sites that differ in extirpation histories. Geochemical signatures of 40 shells, analyzed using atomic absorption spectroscopy, indicated a strong longitudinal pattern. All shells from the two upstream sites had low Hg concentrations (<5-31 microg/kg), shells directly below Saltville had variable, but dramatically higher concentrations (23 4637 microg/kg), and shells from the two downstream sites displayed intermediate Hg levels (<5-115 microg/kg) that declined with distance from Saltville. Two pre industrial shells, collected at Saltville in 1917, yielded very low Hg estimates (5-6 microg/kg). Hg signatures were consistent among mussel species, suggesting that Hg concentrations were invariant to species type; most likely, highly variable Hg levels, both across sites and through time, overwhelmed any interspecific differences in Hg acquisition. Also, a notable postmortem incorporation of Hg in mussel shells seemed unlikely, as the Hg content was not correlated with shell taphonomy (r= 0.18; p = 0.28). The taphonomic analysis (n = 366) showed that the degree of shell alteration reliably distinguished sites with different extirpation histories. At Saltville, where live mussels have been absent for at least 30 years, shells were most heavily altered and fragmented. Conversely, fresh-looking shells abounded upstream, where reproducing mussel populations are still present. In summary, relic shells offered valuable spatio temporal data on Hg concentrations in a polluted ecosystem, and shell taphonomic signatures discriminated sites with different extirpation histories. The shell based strategies exemplified here do not require sampling live specimens and may augment more standard strategies applied to environmental monitoring. The approach should prove especially useful in areas with unknown extirpation and pollution histories. PMID- 15819198 TI - Spatial variation of streamwater chemistry in two Swedish boreal catchments: implications for environmental assessment. AB - To evaluate the scale-dependent spatial variability of water chemistry within two Swedish boreal catchments (subcatchment areas 0.01-78 km2), samples were taken at every junction in the stream network during June 2000 and August 2002. The values of most chemical constituents spanned more than an order of magnitude, and the range was similar to that found in all of Northern Sweden by the national stream survey in 2000. According to the official assessment tools used in Sweden, the entire range of environmental status (for pH, absorbance, alkalinity, dissolved organic carbon (DOC)) and human acidification influence existed within these two study catchments. The water chemistry parameters were relatively stable at catchment areas greater than 15 km2. Sampling at that scale may be adequate if generalized values for the landscape are desired. However the chemistry of headwaters, where much of the stream length and aquatic ecosystem is found would not be characterized. Map parameters correlated to the variability in a key chemical parameter, DOC, but the best predictive map parameters differed markedly between catchments. This study highlights the importance of accounting for headwater spatial variability in environmental assessments of running waters, even in relatively pristine areas. The nature of drainage networks with many headwaters and progressively fewer downstream watercourses makes this a considerable challenge. PMID- 15819199 TI - PCBs and OCPs in sediment cores from the Lower St. Lawrence Estuary, Canada: evidence of fluvial inputs and time lag in delivery to coring sites. AB - Three sediment cores were collected along the longitudinal axis of the Laurentian Trough in the Lower St. Lawrence Estuary (LSLE) and an additional one at the junction of the Estuary and the Gulf of St. Lawrence. After core-slicing, each sediment layer was analyzed for polychlorinated biphenyls (PCBs) and some organochlorine pesticides (OCPs) including p,p'-dichlorodiphenyltrichloroethane (DDT) and its metabolites, hexachlorobenzene (HCB) and Mirex. 210Pb activity was also measured in these sediments, which allowed us to confirm that these cores were too much affected by the overall impact of surface mixing to be dated. Nevertheless, POP sedimentary profiles in cores from the LSLE upstream stations showed well-defined subsurface peak concentrations. Apparently, the peak inputs of POPs to these sediment cores had occurred after the years of maximum sales and production of these chemicals in North America, suggesting a time lag in the delivery of POPs to the LSLE sediments. Concentrations of POPs in the LSLE surface sediments as well as POP inventories in sediment cores decreased in the seaward direction, confirming that the head of the LSLE acts as a sink for sediments and associated constituents. Surface concentrations of sigmaPCBs, sigmaDDTs, and HCB in the most upstream core were on average similar to those reported in two fluvial lakes of the St. Lawrence River but were between 12 and 39 times lower than those from Lake Ontario. For Mirex, the surface concentration in that core was 5 and 130 times lower than the average values found in the fluvial lakes and Lake Ontario, respectively. Differences between Lake Ontario sediment cores and the most upstream core from the LSLE were much smaller on the basis of POP inventories than surface concentrations of POPs, but were still important. The total burdens of POPs in LSLE sediments below the 200 m isobath were 8704 kg for sigmaPCBs, 1825 kg for sigmaDDTs, 319 kg for HCB, and 27.5 kg for Mirex. These values represent between 1% and 10% of the total burdens of these POPs in Lake Ontario sediments. The estimated contribution of POPs by direct atmospheric deposition into the water column area of the LSLE represented at most 30% of the total sediment burdens of POPs in the LSLE, illustrating the importance of fluvial inputs. PMID- 15819200 TI - Molecular characterization of biodegradable dissolved organic matter using bioreactors and [12C/13C] tetramethylammonium hydroxide thermochemolysis GC-MS. AB - Little is known about the molecular composition of the biodegradable fraction of dissolved organic matter (BDOM) in stream ecosystems. We combined plug-flow biofilm reactors, tetramethylammonium hydroxide (TMAH) thermochemolysis GC-MS, and 13C-labeled TMAH thermochemolysis GC-MS to study the molecular composition of BDOM from two stream ecosystems. TMAH products derived from fatty acids, lignin, and other aromatic molecules were quantified using an internal standard approach. We applied the 13C-TMAH thermochemolysis procedure to differentiate between compounds in dissolved organic matter (DOM) that had natural methoxyl groups from those that acquired methoxyl groups during the TMAH reaction. In Rio Tempisquito, a stream draining a tropical evergreen forest, and White Clay Creek, a stream draining a temperate deciduous woodlands, carbohydrates, fatty acids, and lignin contributed to the DOM and BDOM molecular composition. We observed 97 different peaks in the chromatograms of streamwater, with 57% of the peaks common to both streams. The DOM and BDOM pools from each site also contained a unique suite of compounds. Our combined use of TMAH and 13C-TMAH thermochemolysis revealed that heterotrophic bacteria can selectively degrade and demethylate different types of compounds in the lignin residues of DOM. This demonstration of bacterial demethylation of lignin, an abundant and refractory plant molecule, has potential implications for global carbon cycling. PMID- 15819201 TI - Use of transplanted Zebra mussels (Dreissena polymorpha) to assess the bioavailability of microcontaminants in Flemish surface waters. AB - Zebra mussels (Dreissena polymorpha) were translocated in cages to 56 water bodies in Flanders (Belgium) during summer 2001. After six weeks, concentrations of polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), p,p'-DDE, and trace metals were measured in the transplanted mussels. It was investigated whether total dissolved water and sediment pollutant levels or bioaccumulation factors (BAFs) and biota-sediment accumulation factors (BSAFs) were predictive for mussel tissue levels. The sample sites covered a broad range both in terms of the type and concentration of the pollutants, and this was reflected in large differences in tissue concentrations of all pollutants among the sites. The highest pollutant levels in mussels were among the highest reported in the literature. For Cd and Zn levels up to 33 and 1994 microg/g dry wt. respectively were found. The lowest levels were comparable to those from uncontaminated sites in Europe and the U.S. For Cd and Zn respectively 51 and 75% of the variation in tissue levels was described. For both metals, dissolved and particulate metal contributed to the variation in accumulation. For other pollutants, relationships between tissue concentration and water or sediment concentration were weak or nonsignificant. Then the measured environmental factors (dissolved calcium, pH, oxygen, organic carbon and clay content in the sediment) were taken into account applying multiple regression analysis, and no increase in the described variation of pollutant accumulation was observed. The BAF and BSAF for all pollutants varied up to 1000-fold even after TOC-normalization. Clear negative relationships were found between BAFs/ BSAFs and environmental levels. However, even at constant environmental concentrations a 10- to 100-fold variation in BAFs/BSAFs was observed. This study illustrated the need for biological monitoring since neither environmental PMID- 15819202 TI - Colloid stability in vadose zone Hanford sediments. AB - We experimentally determined colloid stability of natural colloids extracted from vadose zone sediments from the U.S. Department of Energy's Hanford Reservation. We also used reference minerals, kaolinite, montmorillonite, and silica,for comparative purposes. Colloid stability was assessed with two different methods: the batch turbidity method and dynamic light scattering. Critical coagulation concentrations (CCCs) were determined for pure Na and pure Ca electrolyte solutions, as well for mimicked Hanford vadose zone pore waters with varying sodium adsorption ratios (SARs). Critical coagulation concentrations obtained from the batch turbidity method were sensitive to initial colloid mass concentrations, settling time, and CCC criteria. The lower the initial colloid concentration and the shorter the settling times were, the larger was the CCC. The CCCs determined from the dynamic light scattering, where diluted colloidal suspensions are used, were not dependent on settling time and arbitrary CCC criteria, so dynamic light scattering is therefore the preferred method to determine colloid stability. The CCC values determined from dynamic light scattering ranged from 90 to 200 mmol/L for Na systems and 1.7 to 3.8 mmol/L for Ca systems. The stability of natural colloids was intermediate between that of pure kaolinite and montmorillonite. The results indicate that colloids in the Hanford vadose zone form stable suspensions, i.e., are in the slow aggregation regime. Nonetheless, due to the long travel times in the vadose zone, nearly all colloids will aggregate and be removed from the water column before reaching groundwater levels. PMID- 15819203 TI - Growth dilution of metals in microalgal biofilms. AB - Despite the key role microalgae play in introducing toxicants into aquatic food webs, little is known about the effects of environmental factors on metal accumulation by these primary producers. Environmental factors such as light and nutrients alter growth rates and may consequently influence metal concentrations in microalgae through growth dilution. Laboratory experiments suggested that metal uptake and elimination by microalgal biofilms were gradual enough to enable dilution of metals within the biofilms by photosynthetically accrued carbon, and a simple kinetic model of metal accumulation predicted significant variation in metal content due to growth dilution over the natural range of microalgal growth rates. The ratio of metal uptake to carbon uptake by microalgal biofilms decreased exponentially with increasing light in short-term laboratory experiments because photosynthesis was much more sensitive to a light gradient than was metal uptake. The effect of light on biofilm metal concentrations was confirmed in situ with a long-term experiment in which experimental shading of biofilms in a metal-contaminated stream decreased biofilm growth rates and caused a 3x increase in biofilm concentrations of twelve metals, including methylmercury. Slow growth at the primary producer level is a likely contributor to higher biotic metal concentrations in shaded, oligotrophic, or cold ecosystems. PMID- 15819204 TI - Evidence for anthropogenic 210Po in the urban atmosphere of Seoul, Korea. AB - We have measured the concentrations of 210Po, 210Pb, SO4(2-), Na+, and 34S in precipitation samples from two metropolitan cities, Seoul and Busan, Korea. The delta34S values ranged from 0 to 10% in most Seoul and Busan precipitation samples, indicating major contributions from industrial sources to S levels. A high level of excess 210Po, which is not produced by 210Pb decay in the troposphere, was observed in both regions. The excess 210Po activities in some samples from Busan, a harbor city, were influenced strongly by sea salt (i.e., sea-surface microlayer) which could be traced using [Na+] and delta34S values. In Seoul precipitations, we observed a good correlation between non-sea-salt SO4(2-) and excess 210Po, suggesting that both species are controlled mainly by the same factor. This correlation and the delta34S values indicate that the major source for both species in this region is likely to be anthropogenic, rather than from traditionally suggested sources such as soil resuspension, stratospheric air intrusion, sea sprays, volcanic emissions, and biogenic emissions. PMID- 15819205 TI - Occurrence and solid-liquid partition of sulfonated naphthalene-formaldehyde condensates in the aquatic environment. AB - Sulfonated naphthalene-formaldehyde condensates (SNFC) are high production volume chemicals used in a variety of applications, for example, as concrete plasticizers, tanning agents, or dye dispersants. They enter the aquatic environment primarily by the wastewater path. The occurrence and fate of the monomers, which are different isomers of mono- and disulfonated naphthalene, was intensively investigated in previous studies. However, the environmental fate of the persistent higher molecular SNFC is so far widely unknown. This paper describes an ultrasonic extraction under alkaline conditions, followed by ion pair HPLC with fluorescence detection for the analysis of SNFC oligomers from solid environmental matrixes such as sewage sludge, suspended solids, and river sediments. Limits of quantification of about 0.1 mg kg-1 d.m. were well below the measured concentrations in environmental samples. SNFC were adsorbed to suspended solids and river sediments in three major German rivers (Rhine, Neckar, and Danube) in concentrations typically up to several mg kg(-1) d.m. A total content of about 4 g kg(-1) d.m. was measured in a sewage sludge of a municipal wastewater treatment plant, which receives wastewater from a textile dyeing plant. Furthermore, the first quantitative field data on the partition of SNFC and their monomers between the aqueous phase and solid environmental compartments are presented. Solid-liquid partition coefficients (Kd) of oligomers with a chain length ranging from three to six naphthalenesulfonate units were derived from the analysis of corresponding wastewater and sewage sludge samples and from suspended solids and river water samples, respectively. Determined Kd values were in the range from 10(2) to 10(4) L kg(-1). PMID- 15819206 TI - Heterogeneous reactions of glyoxal on particulate matter: identification of acetals and sulfate esters. AB - Reactive uptake of glyoxal onto particulate matter has been studied in laboratory experiments in a 2 m3 Teflon reaction chamber. Inorganic seed particles of different composition were utilized, including (NH4)2SO4, (NH4)2SO4/ H2SO4, NaNO3, and simulated sea salt, while the relative humidity and acid concentration were varied. The organic composition of the growing particles was measured in situ with an aerosol mass spectrometer, providing particle mass spectra as a means of product identification. Aerosol physical characteristics were also measured with a differential mobility analyzer and condensation nucleus counter. Regardless of seed composition, particle growth was rapid and continuous over the course of several hours. Identification of several mass fragments greater than the glyoxal monomer suggested that heterogeneous reactionsto form glyoxal adducts of lowvolatility had occurred. Temporal analysis of the mass fragments was consistent with a proposed acid-catalyzed mechanism whereby glyoxal is first hydrated, followed by self-reaction to form cyclic acetal structures. Increased relative humidity slowed the formation of higher order oligomers, also consistent with the proposed mechanism. The relative contribution of various oligomers to the overall organic composition was strongly dependent on the relative humidity and hence the particulate water concentration. A mild acid catalysis was also observed upon increasing the acidity of the seed particles. Specific mass fragments were found that could only arise from sulfate esters and were not present on the non-sulfur-containing seed particles. This first evidence of the formation of organic sulfates in particles is presented together with a proposed mechanism and molecular structure. These results suggest that the formation of these products of glyoxal uptake can contribute significantly to secondary organic aerosol. PMID- 15819207 TI - Copper toxicity to larval stages of three marine invertebrates and copper complexation capacity in San Diego Bay, California. AB - Temporal and spatial measurements of the toxicity (EC50), chemical speciation, and complexation capacity (Cu-CC) of copper in waters from San Diego Bay suggest control of the Cu-CC over copper bioavailability. While spatial distributions of total copper (CuT) indicate an increase in concentration from the mouth toward the head of San Diego Bay, the distribution of aqueous free copper ion (Cu(II)aq) shows the opposite trend. This suggests that the bioavailability of copper to organisms decreases toward the head of the bay, and is corroborated by the increase in the amount of copper needed to reach an EC50, observed for larval stages of three marine invertebrates (Mediterranean mussel, Mytilus galloprovincialis, sand dollar, Dendraster excentricus, and purple sea urchin, Strongylocentrotus purpuratus), and by the increase in Cu-CC heading into the head of the bay. The amount of Cu(II)aq required to produce a 50% reduction in normal larval development (referred to here as pCuTox,) of the mussel, the most sensitive of the three marine invertebrates, was generally at or above approximately 1 x 10(-11) mol L(-1) equivalents of Cu (i.e., pCuTox approximately 11 = -(log [Cu(II)aq])). These results suggest that the copper complexation capacity in San Diego Bay controls copper toxicity by keeping the concentration of Cu(II)aq at nontoxic levels. PMID- 15819208 TI - Speciation of mercury and mode of transport from placer gold mine tailings. AB - Historic placer gold mining in the Clear Creek tributary to the Sacramento River (Redding, CA) has highly impacted the hydrology and ecology of an important salmonid spawning stream. Restoration of the watershed utilized dredge tailings contaminated with mercury (Hg) introduced during gold mining, posing the possibility of persistent Hg release to the surrounding environment, including the San Francisco Bay Delta. Column experiments have been performed to evaluate the extent of Hg transport under chemical conditions potentially similar to those in river restoration projects utilizing dredge tailings such as at Clear Creek. Physicochemical perturbations, in the form of shifts in column influent ionic strength and the presence of a low molecular weight organic acid, were applied to coarse and fine sand placer tailings containing 109-194 and 69-90 ng of Hg/g, respectively. Significant concentrations of mercury, up to 16 microg/L, leach from these sediments in dissolved and particle-associated forms. Sequential chemical extractions (SCE) of these tailings indicate that elemental Hg initially introduced during gold mining has been transformed to readily soluble species, such as mercury oxides and chlorides (3-4%), intermediately extractable phases that likely include (in)organic sorption complexes and amalgams (75-87%), and fractions of highly insoluble forms such as mercury sulfides (6-20%; e.g., cinnabar and metacinnabar). Extended X-ray absorption fine structure (EXAFS) spectroscopic analysis of colloids obtained from column effluent identified cinnabar particles as the dominant mobile mercury-bearing phase. The fraction of intermediately extractable Hg phases also likely includes mobile colloids to which Hg is adsorbed. PMID- 15819209 TI - Hygroscopicity of water-soluble organic compounds in atmospheric aerosols: amino acids and biomass burning derived organic species. AB - Amino acids and organic species derived from biomass burning can potentially affect the hygroscopicity and cloud condensation activities of aerosols. The hygroscopicity of seven amino acids (glycine, alanine, serine, glutamine, threonine, arginine, and asparagine) and three organic species most commonly detected in biomass burning aerosols (levoglucosan, mannosan, and galactosan) were measured using an electrodynamic balance. Crystallization was observed in the glycine, alanine, serine, glutamine, and threonine particles upon evaporation of water, while no phase transition was observed in the arginine and asparagine particles even at 5% relative humidity (RH). Water activity data from these aqueous amino acid particles, except arginine and asparagine, was used to revise the interaction parameters in UNIQUAC functional group activity coefficients to give predictions to within 15% of the measurements. Levoglucosan, mannosan, and galactosan particles did not crystallize nor did they deliquesce. They existed as highly concentrated liquid droplets at low RH, suggesting that biomass burning aerosols retain water at low RH. In addition, these particles follow a very similar pattern in hygroscopic growth. A generalized growth law (Gf = (1 - RH/100)-0.095) is proposed for levoglucosan, mannosan, and galactosan particles. PMID- 15819210 TI - Vapor-phase exchange of perchloroethene between soil and plants. AB - Tree core concentrations of tetrachloroethylene (perchloroethene, PCE) at the Riverfront Superfund Site in New Haven, MO, were found to mimic the profile of soil phase concentrations. The observed soil-tree core relationship was stronger than that of groundwater PCE to tree core concentrations atthe same site. Earlier research has shown a direct, linear relationship between tree core and groundwater concentrations of chlorinated solvents and other organics. Laboratory scale experiments were performed to elucidate this phenomenon, including determining partitioning coefficients of PCE between plant tissues and air and between plant tissues and water, measured to be 8.1 and 49 L/kg, respectively. The direct relationship of soil to tree core PCE concentrations was hypothesized to be caused by diffusion between tree roots and the soil vapor phase in the subsurface. The central findings of this research are discovering the importance of subsurface vapor-phase transfer for VOCs and uncovering a direct relationship between soil vapor-phase chlorinated solvents and uptake rates that impact contaminant translocation from the subsurface and transfer into the atmosphere. PMID- 15819211 TI - The origin of naturally occurring perchlorate: the role of atmospheric processes. AB - Perchlorate, an iodide uptake inhibitor, is increasingly being detected in new places and new matrices. Perchlorate contamination has been attributed largelyto the manufacture and use of ammonium perchlorate (the oxidizer in solid fuel rockets) and/or the earlier use of Chilean nitrate as fertilizer (approximately 0.1% perchlorate). However, there are regions such as the southern high plains (Texas Panhandle) where there is no clear historical or current evidence of the extensive presence of rocket fuel or Chilean fertilizer sources. The occurrence of easily measurable concentrations of perchlorate in such places is difficult to understand. In the southern high plains groundwater, perchlorate is better correlated with iodate, known to be of atmospheric origin, compared to any other species. We show that perchlorate is readily formed by a variety of simulated atmospheric processes. For example, it is formed from chloride aerosol by electrical discharge and by exposing aqueous chloride to high concentrations of ozone. We report that perchlorate is present in many rain and snow samples. This strongly suggests that some perchlorate is formed in the atmosphere and a natural perchlorate background of atmospheric origin should exist. PMID- 15819212 TI - Organic complexation of Fe(II) and its impact on the redox cycling of iron in rain. AB - More than 80% of the iron(II) present in a dilute (pH 4.5) H2SO4 solution was oxidized by hydrogen peroxide (3 microM) in 24 h, whereas in rainwater Fe(II) remained stable for days indicating that a complexed form of Fe(II) exists in rainwater that protects it against oxidation. When a rain sample was irradiated for 2 h with simulated sunlight, there was a 57 nM increase in Fe(II) resulting from photoreduction of organic Fe(III) complexes. Once irradiation ceased, the photoproduced Fe(II) rapidly oxidized back to its initial concentration of 32 nM prior to irradiation, but not to zero. These photochemical studies demonstrate that during the daytime when sunlight is present there are dynamic interconversions between complexed and uncomplexed Fe(II) and Fe(III) species in rainwater. During the night, after the photochemically produced Fe(II) is reoxidized to Fe(III), virtually all remaining Fe(II) is complexed by ligands which resist further oxidation. Rain samples oxidized under intense UV light lost their ability to stabilize Fe(II), suggesting the ligands stabilizing Fe(II) are organic compounds destroyed by UV-irradiation. Additional UV-irradiation studies demonstrated that on average 25% of the Fe-complexing ligands in rainwater are extremely strong and cannot be detected by spectrophotometric analysis using ferrozine. The stability of organically complexed Fe(II) has important implications for the bioavailability of rainwater-derived Fe in the surface ocean. PMID- 15819213 TI - Bioaerosol emission rate and plume characteristics during land application of liquid class B biosolids. AB - This study investigated bioaerosol emission rates and plume characteristics of bioaerosols generated during land application of liquid Class B biosolids. In addition, it compared the rate of aerosolization of coliphages and total coliform bacteria during land application of liquid Class B biosolids to the rate of aerosolization during land application of groundwater inoculated with similar concentrations of Escherichia coli and coliphage MS2. Air samples were taken immediately downwind of a spray applicator as it applied liquid (approximately 8% solids) biosolids to farmland near Tucson, Arizona. Air samples were also collected immediately downwind of groundwater seeded with MS2 and E. coli applied to land in an identical manner. Air samples, collected with liquid impingers, were taken in horizontal and vertical alignment with respect to the passing spray applicator. Vertical and horizontal sample arrays made it possible to calculate the flux of microorganisms through a virtual plane of air samplers, located 2 m downwind of the passing spray applicator. Neither coliphages nor coliform bacteria were detected in air downwind of spray application of liquid Class B biosolids. Based on limits of detection for the methodology, the rate of aerosolization during land application of liquid biosolids was calculated to be less than 33 plaque forming units (PFU) of coliphage and 10 colony forming units (CFU) of coliform bacteria per meter traveled by the spray applicator. The rate of aerosolization during land application of seeded groundwater was found to be, on average, 2.02 x 10(3) CFU E. coli and 3.86 x 10(3) PFU MS2 aerosolized per meter traveled by the spray applicator. This is greater aerosolization than was observed during land application of biosolids. Because concentrations of coliphages and coliforms were similar in the liquid biosolids and the seeded water, itwas concluded that some property of biosolids reduces aerosolization of microorganisms relative to groundwater. Additional experiments utilizing a novel air sampling protocol showed that the duration of bioaerosol exposure immediately (2 m) downwind of biosolids spray application is brief and the plume of bioaerosols generated is discrete. Additional air samples showed that aerosolization of coliphages and coliform bacteria after liquid biosolids have been applied to land does not occur at detectable levels. PMID- 15819214 TI - Desorption kinetics of Cd, Zn, and Ni measured in soils by DGT. AB - DGT (diffusive gradients in thin films) was used to measure the distribution and rates of exchange of Zn, Cd, and Ni between solid phase and solution in five different soils. Soil texture ranged from sandy loam to clay, pH ranged from 4.9 to 7.1, and organic carbon content ranged from 0.8% to 5.8%. DGT devices continuously remove metal to a Chelex gel layer after passage through a well defined diffusion layer. The magnitude of the induced remobilization flux from the solid phase is related to the pool size of labile metal and the exchange kinetics between dissolved and sorbed metal. DGT devices were deployed over a series of times (4 h to 3 weeks), and the DIFS model (DGT induced fluxes in soils) was used to derive distribution coefficients for labile metal (Kdl) and the rate at which the soil system can supply metal from solid phase to solution, expressed as a response time. Response times for Zn and Cd were short generally (<8 min). They were so short in some soils (<1 min) that no distinction could be made between supply of metal being controlled by diffusion or the rate of release. Generally longer response times for Ni (5-20 min) were consistent with its slow desorption. The major factor influencing Kdl for Zn and Cd was pH, but association with humic substances in the solid phase also appeared to be important. The systematic decline, with increasing pH, in both the pool size of Ni available to the DGT device and the rate constant for its release is consistent with a part of the soil Ni pool being unavailable within a time scale of 1-20 min. This kinetic limitation is likely to limit the availability of Ni to plants. PMID- 15819215 TI - Remediation of polycyclic aromatic hydrocarbon compounds in groundwater using poplar trees. AB - A seven-year study was conducted to assess the effectiveness of hybrid poplar trees to remediate polycyclic aromatic hydrocarbon (PAH) compounds in soil and groundwater at a creosote-contaminated site. A reduction in the areal extent of the PAH plume was observed in the upper half of the 2-m-thick saturated zone, and PAH concentration levels in the groundwater declined throughout the plume. PAH concentrations began to decline during the period between the third and fourth growing seasons, which coincided with the propagation of the tree roots to the water table region. Remediation was limited to naphthalene and several three-ring PAHs (acenaphthylene and acenaphthene). PAH concentrations in soil and aquifer sediment samples also declined over time; however, levels of four-ring PAHs persisted at the lower depths during the study period. The naphthalene to total PAH concentration ratio in the most contaminated groundwater decreased from >0.90 at the beginning of the second growing season to approximately 0.70 at the end the study. Remediation in the lower region of the saturated zone was limited bythe presence of a 0.3-m-thick layer of creosote present as a dense nonaqueous phase liquid (DNAPL). The nearly steady-state condition of the PAH concentrations observed during the last three years of the study suggests that the effectiveness of the phytoremediation system is limited by the rate of PAH dissolution from the DNAPL source. PMID- 15819216 TI - Characterization of charcoal adsorption sites for aromatic compounds: insights drawn from single-solute and bi-solute competitive experiments. AB - Charcoal, the residue of incomplete biomass burning that is found in many soils and sediments, is considered a high affinity sorbent for organic pollutants. However, little is known about the microscopic processes controlling sorption. The purpose of this study was to gain molecular-scale insight into the sorption on a charcoal of three weakly soluble aromatic compounds [benzene (BEN), toluene (TOL), and nitrobenzene (NBZ)] by conducting both single-solute and bi-solute experiments. The charcoal (420 m2 g(-1)) was produced from maple wood shavings by oxygen-limited pyrolysis at 673 K. Solute affinity for charcoal followed the order NBZ > TOL > BEN. Commonly employed sorption models did not adequately describe the single-solute isotherms. Competition in both TOL-BEN and the TOL-NBZ bi-solute systems was strong. Normalization of the isotherms for the hydrophobic driving force by using an existing free energy correlation between sorption and partitioning to an inert solvent (benzene or n-hexadecane) with a nonpolar aromatic compound calibration set resulted in a finding of enhanced sorption of NBZ relative to the coalesced BEN and TOL isotherms, indicating some specificity in the interaction of NBZ. The competitive data indicated 1:1 molar competition between BEN and TOL and between NBZ and TOL, showing conclusively that this specificity was not due to a subpopulation of sorption sites unique to NBZ. H bonding was ruled out, as the relative affinity for the sorbent among the solutes did not change at all when increasing the solution pH from 6.5 to 11. 1H NMR experiments showed molecular complexation in chloroform between NBZ and model graphene polycyclic aromatic units (naphthalene, phenanthrene, and pyrene) which was absentfor BEN and TOL. This result, in combination with the results of a companion study (Zhu and Pignatello, Environ. Sci. Technol. (in press)), is used to support the existence of pi-pi electron donor-acceptor interactions between NBZ (electron acceptor) and the polycyclic aromatic charcoal surface (electron donor) as the cause of enhanced NBZ sorption. PMID- 15819217 TI - Biodegradability of fractions of dissolved organic carbon leached from decomposing leaf litter. AB - Dissolved organic matter leached from decomposing organic matter is important in the leaching of nutrients from the root zone of ecosystems, eluviation of metals, and transport of hydrophobic pollutants. The objective of this study was to compare microbial mineralization rates in intact soil cores of various fractions of water-soluble dissolved organic matter. Uniformly 14C-labeled Populus fremontii leaf litter that had decomposed for 1 year was extracted in water and this extract was fractionated into phenolic, humic acid, fulvic acid, hydrophilic acid, and hydrophilic neutral fractions. Fulvic acid comprised 42.1% of C in dissolved organic carbon (DOC) extracted from the litter. These fractions were added to intact cores of soil or sand, and respired 14CO2 was collected. The percentage of labeled substrate C mineralized in soil at the end of 1 year was, in order from least to greatest, hydrophilic acid (30.5), fulvic acid (33.8), humic acid (39.0), whole, unfractionated DOC (43.5), unseparated hydrophilic acid and neutral (44.7), phenolic (63.3), glucose (66.4), and hydrophilic neutral (70.2). In acid-washed nutrient-amended sand that was inoculated with soil microbes, mineralization rates of fulvic acid and glucose were lower. The fractionation appeared to separate the DOC into components with widely different rates of mineralization. Results also supported the ideas that the dissolved humic substance and hydrophilic acid fractions are inherently difficult for microbes to mineralize, and this property can contribute to movement of refractory C in soil and into aquatic ecosystems. PMID- 15819218 TI - Solubility of hydrophobic organic pollutants in binary and multicomponent aqueous solvents. AB - The present paper deals with the application of fluctuation theory of solutions to the solubility of poorly soluble substances of environmental significance in aqueous mixed solvents. The fluctuation theory of ternary solutions was first used to derive an expression for the activity coefficient of a solute at infinite dilution in a binary mixed solvent. This equation contains the activity coefficients of the constituents of the solute-free mixed solvent and the molar volume of the solute-free mixed solvent. Further, the derived expression for the activity coefficient of a solute at infinite dilution was used to generate a number of expressions for the solubility of solids in aqueous mixed solvents. Several expressions for the activity coefficients of the components were considered: first, the mixed solvent was considered an ideal mixture; second, the activity coefficients of the constituents of the binary solvent were expressed using the two-suffix Margules equations; third, the activity coefficients of the constituents of the binary solvent were expressed using the Wilson equations. The obtained expressions were applied to 25 experimental data sets pertaining to the solubilities of hydrophobic organic pollutants (HOP) in aqueous mixed solvents. It was found that the suggested equations can be used for an accurate and reliable correlation of the solubilities in aqueous mixed binary solvents. The best results were obtained by combining our expression for the activity coefficient of a solute at infinite dilution in a mixed solvent with the Wilson equations for the activity coefficients of the constituents of a solute-free mixed solvent. The derived equations can also be used for predicting the solubilities of poorly soluble environmentally important compounds in aqueous mixed solvents using for the Wilson parameters those obtained from vapor-liquid equilibrium data. A similar methodology was applied to the solubility of poorly soluble substances of environmental significance in multicomponent (ternary and higher) aqueous mixed solvents. The expression for the activity coefficient of a solute in an ideal multicomponent mixed solvent was used to derive an equation for the solubility of a poorly soluble solute in an ideal multicomponent mixed solvent in terms of its solubilities in two subsystems of the multicomponent solvent and their molar volumes. Ultimately the solubility could be expressed in terms of those in binary or even in the individual constituents of the solvent and their molar volumes. The computational method was applied to predict the solubilities of naphthalene and anthracene in ternary, quaternary and quinary aqueous mixed solvents. The results were compared with experiment and good agreement was obtained. PMID- 15819219 TI - Atmospheric chemistry of hydrazoic acid (HN3): UV absorption spectrum, HO reaction rate, and reactions of the N3 radical. AB - Processes related to the tropospheric lifetime and fate of hydrazoic acid, HN3, have been studied. The ultraviolet absorption spectrum of HN3 is shown to possess a maximum near 262 nm with a tail extending to at least 360 nm. The photolysis quantum yield for HN3 is shown to be approximately 1 at 351 nm. Using the measured spectrum and assuming unity quantum yield throughout the actinic region, a diurnally averaged photolysis lifetime near the earth's surface of 2-3 days is estimated. Using a relative rate method, the rate coefficient for reaction of HO with HN3 was found to be (3.9 +/-0.8) x 10(-12) cm3 molecule(-1) s(-1), substantially larger than the only previous measurement. The atmospheric HN3 lifetime with respect to HO oxidation is thus about 2-3 days, assuming a diurnally averaged [HO] of 10(6) molecule cm(-3). Reactions of N3, the product of the reaction of HO with HN3, were studied in an environmental chamber using an FTIR spectrometer for end-product analysis. The N3 radical reacts efficiently with NO, producing N2O with 100% yield. Reaction of N3 with NO2 appears to generate both NO and N2O, although the rate coefficient for this reaction is slower than that for reaction with NO. No evidence for reaction of N3 with CO was observed, in contrast to previous literature data. Reaction of N3 with O2 was found to be extremely slow, k < 6 x 10(-20) cm3 molecule(-1) s(-1), although this upper limit does not necessarily rule out its occurrence in the atmosphere. Finally, the rate coefficient for reaction of Cl with HN3 was measured using a relative rate method, k = (1.0+/-0.2) x 10(-12) cm3 molecule(-1) s(-1). PMID- 15819220 TI - Influence of addition order and contact time on thorium(IV) retention by hematite in the presence of humic acids. AB - The influence of addition order and contact time in the system hematite (alpha Fe2O3)-humic acid (HA)-thorium(IV) (Th(IV)) was studied in batch experiments. Th(IV) is considered here as a chemical analogue of other actinides (IV). The sorption isotherms were acquired varying pH in the range 2-10 and HA concentration in the range 1-100 mg/L. As already observed by numerous authors, Th(IV) retention was hindered when HA and hematite were equilibrated beforehand during 24 h. As it has been observed in a previous study, this effect was drastic when the ratio between humic and surface (iron oxide) sites exceeds a critical value. However, when HA was added after a 24-h equilibration of the hematite Th(IV) system, Th(IV) was barely desorbed from the iron oxide surface. Furthermore, no drastic effect of the ratio between humic and surface sites could be evidenced, as the increase of HA concentration only results in a slight monotonic decrease in Th(IV) retention. Increasing contact time between components of the systems only indicated slight Th(IV) retention variation. This was interpreted as a consequence of slow kinetic controls of both the Th(IV)-HA complexation and HA-hematite sorption. PMID- 15819221 TI - Sorption kinetics of organic contaminants by sandy aquifer and its kerogen isolate. AB - Long-term sorption behaviors of phenanthrene (Phen) on the Borden sand from 1 min to 365 days and of Phen and 1,2-dichlorobenzene (DCB) on the isolated kerogen from 1 to 120 days were characterized by examining the time dependence of solute phase distribution relationships (PDRs) and compared with the prior reported sorption of tetrachlorobenzene (TeCB) and tetrachloroethene (PCE) on the pulverized and/or acid-treated bulk sand and size fractions. The sorption kinetics for Phen on the bulk sand and its kerogen isolate can well be described by the fractional power kinetics equation (q(e) = kt(b)). The similar rate parameter b for q(e)(t) vs t at 5-7 levels of initial concentrations of Phen on the two sorbents, respectively, ranging from 0.077 to 0.099 and from 0.072 to 0.086, indicates the similar sorption kinetics rate. The modified-Freundlich parameters of TeCB on the pulverized or acid-treated 0.3-mm size fraction match those of Phen on the isolated kerogen, suggesting the same natural organic matter (NOM) property of the two sorbents. As the prior investigation underestimated the Koc value for the TeCB sorption on the acid-treated 0.3-mm size fraction by a factor of 1.76, the estimated time to reach 95% of sorption equilibrium is much longer than the prior estimation (over 10 years vs about 2.5 years). The estimated times to reach 95% of sorption equilibrium at three levels of relative solubility for Phen on the bulk sand and its isolated kerogen are, respectively, longer than one decade, demonstrating the similar diffusion length for Phen on the two sorbents. The observed slow sorption kinetics is related to nanometer pore diffusion within kerogen matrix. The investigation supplies new clues for explaining the often observed much longer persistence of organic contaminants in soils and sediments than the prediction based on the short-term laboratory experiment. PMID- 15819222 TI - Nonmonotonic variations in deposition rate coefficients of microspheres in porous media under unfavorable deposition conditions. AB - The transport of carboxylate-modified polystyrene latex microspheres was examined in packed quartz sand under a variety of environmentally relevant ionic strength and flow conditions. The retained concentrations of microspheres in the sediment increased first, and then decreased with transport distance, indicating that the deposition rate coefficient changed nonmonotonically over the transport distance. This finding demonstrates the ubiquity of spatial variation in deposition rate coefficients under unfavorable deposition conditions, and in addition indicates that the previously recognized monotonic decrease with transport distance is not the sole form of spatial variations in deposition rate coefficients. In contrast, the deposition rate coefficients of similarly sized microspheres with different surface group densities were shown to decrease monotonically with transport distance in the same porous media, indicating that the form of spatial variation in deposition rate coefficient is highly sensitive to system conditions. The ubiquity and sensitivity of the spatial variation of deposition rate coefficients indicate that current practices that utilize log-linear extrapolation of discreet measurements of colloid attenuation to determine colloid removal with distance from source are not valid (for both biological and nonbiological colloids). The retained colloid profiles hold the promise to reveal processes governing colloid deposition under unfavorable conditions that are yet to be identified. PMID- 15819223 TI - Effect of algal and bacterial diet on methyl mercury concentrations in zooplankton. AB - We studied the effect of zooplankton diet on MeHg accumulation in different zooplankton size-fractions from lakes of different trophic status. Using fatty acid biomarkers, we tested the hypotheses that (a) variations of MeHg concentrations are determined bythe taxonomic composition of zooplankton and (b) concentrations of dietary algal and bacterial compounds can predict MeHg concentrations of seston (10-64 microm), micro- (100-200 microm), meso- (200-500 microm), and macrozooplankton (>500 microm) in lakes on Vancouver Island, Canada. MeHg concentrations increased from seston (4-48 ng g dry weight(-1)) to macrozooplankton (94-240 ng g dry weight(-1)), indicating that MeHg accumulated as a function of plankton size. Results from linear regression analysis showed that MeHg concentrations were not significantly related to the taxonomic composition of zooplankton. However, using dietary lipid biomarkers, we demonstrated that bacterial diet (R2 = 0.50; p < 0.01) could better predict variations of MeHg concentrations in zooplankton than essential algal diet (R2 = 0.35; p < 0.01). Because MeHg accumulation within the planktonic food web was higher (20x) than the observed accumulation of total bacterial (6.5x) and algal (4.7x) diet biomarkers, zooplankton retained dietary MeHg more efficiently than bacterial and algal diet compounds. These results indicate that MeHg of macrozooplankton, the preferred prey size of planktivorous fish, is more efficiently transferred than essential diet compounds to organisms at higher trophic levels. PMID- 15819224 TI - Protein nitration by polluted air. AB - The effects of air pollution on allergic diseases are not yetwell-understood. Here, we show that proteins, in particular birch pollen proteins including the allergen Bet v 1, are efficiently nitrated by polluted air. This posttranslational modification of proteins is likely to trigger immune reactions and provides a molecular rationale for the promotion of allergies bytraffic related air pollution. Enzyme immunoassays have been used to determine equivalent degrees of nitration (EDN) for protein samples exposed to urban outdoor air and synthetic gas mixtures. The observed rates of nitration were governed by the abundance of nitrogen oxides and ozone, and concentration levels typical for summer smog conditions led to substantial nitration within a few hours to days (EDN up to 20%). Moreover, nitrated proteins were detected in urban road dust, window dust, and fine air particulate matter (EDN up to 0.1%). PMID- 15819225 TI - Determination of mercury evasion in a contaminated headwater stream. AB - Evasion from first- and second-order streams in a watershed may be a significant factor in the atmospheric recycling of volatile pollutants such as mercury; however, methods developed for the determination of Hg evasion rates from larger water bodies are not expected to provide satisfactory results in highly turbulent and morphologically complex first- and second-order streams. A new method for determining the Hg evasion rates from these streams, involving laboratory gas indexing experiments and field tracer tests, was developed in this study to estimate the evasion rate of Hg from Gossan Creek, a first-order stream in the Upsalquitch River watershed in northern New Brunswick, Canada. Gossan Creek receives Hg-contaminated groundwater discharge from a gold mine tailings pile. Laboratory gas-indexing experiments provided the ratio of gas-exchange coefficients for zero-valent Hg to propane (tracer gas) of 0.81+/-0.16, suggesting that the evasion mechanism in highly turbulent systems can be described by the surface renewal model with an additional component of enhanced gas evasion probably related to the formation of bubbles. Deliberate field tracer tests with propane and chloride tracers were found to be a reliable and practical method for the determination of gas-exchange coefficients for small streams. Estimation of Hg evasion from the first 1 km of Gossan Creek indicates that about 6.4 kg of Hg per year is entering the atmosphere, which is a significant fraction of the regional sources of Hg to the atmosphere. PMID- 15819226 TI - Sorption of ionic surfactants to estuarine sediment and their influence on the sequestration of phenanthrene. AB - The sorption of an anionic surfactant (sodium dodecyl sulfate; SDS) and a cationic surfactant (hexadecyl trimethylammonium bromide; HDTMA) to estuarine sediment has been studied in river water and seawater. Sorption isotherms for SDS were essentially linear in both waters, suggesting a nonspecific, hydrophobic interaction between the SDS tail and particle surface. Sorption of HDTMA was considerably greater, more nonlinear, and more sensitive to water composition. These observations were attributed to a combination of both electrostatic and hydrophobic interactions between the surfactant and particle surface, the formation of admicelles, and salinity-induced structural alteration of the hydrophobic tail of the HDTMA molecule. Presence of SDS caused a reduction in the sorption of phenanthrene to estuarine sediment because of the competitive effects of the surfactant tail for hydrophobic sorption sites on the particle surface. Conversely, the presence of HDTMA caused significant enhancement in phenanthrene sequestration because of head-on sorption of surfactant molecules and a resulting, more hydrophobic particle surface. The most persistent feature of our results was an inverse dependence of unit sorption on particle concentration, and an empirical algorithm defining the effect was used to calculate the sediment water fractionation of realistic concentrations of reactants in the estuarine water column. The results of these calculations, and the more general findings of this study, significantly improve our understanding of both the transport and fate of ionic surfactants in the estuarine environment, and the effects that these surfactants have on the partitioning of hydrophobic organic micropollutants. PMID- 15819227 TI - Dissolution-induced contact angle modification in dense nonaqueous phase liquid/water systems. AB - The contact angle between DNAPL, water, and aquifer material interfaces influences the spatial distribution of DNAPLs as they infiltrate into the aquifer, and may ultimately influence their remediation. The objective of this work was to evaluate the effects of dissolution on contact angle. Just as physically retracting a sessile drop reduces its contact angle with a surface, it was speculated that dissolution could cause contact angles to be reduced. Long term dissolution experiments were conducted over the course of days to weeks, examining the dissolution of sessile drops of two DNAPLs, trichloroethylene (TCE) and tetrachloroethylene (PCE), in water and low concentration surfactant solutions, on glass surfaces. Experiments found that dissolution led to a continuous decrease of contact angle measured through the DNAPL drop, in most cases to near 0 degrees, far lower than angles achievable through measurements of receding contact angles for the same systems. Pinning of drop contact diameter was observed in most experiments. A model developed on the basis of the Bashforth Adams equation to predict the effect of dissolution on contact angle for drops with a pinned contact diameter showed very good agreement with experimental observations. PMID- 15819228 TI - Use of personal-indoor-outdoor sulfur concentrations to estimate the infiltration factor and outdoor exposure factor for individual homes and persons. AB - A study of personal, indoor, and outdoor exposure to PM2.5 and associated elements has been carried out for 37 residents of the Research Triangle Park area in North Carolina. Participants were selected from persons expected to be at elevated risk from exposure to particles, and included 29 persons with hypertension and 8 cardiac patients with implanted defibrillators. Participants were monitored for 7 consecutive days in each of four seasons. One goal of the study was to estimate the contribution of outdoor PM2.5 to indoor concentrations. This depends on the infiltration factor Finf, the fraction of outdoor PM2.5 remaining airborne after penetrating indoors. After confirming with our measurements the findings of previous studies that sulfur has few indoor sources, we estimated an average Finf for each house based on indoor/outdoor sulfur ratios. These estimates ranged from 0.26 to 0.87, with a median value of 0.55. Since these estimates apply only to particles of size similar to that of sulfur particles (0.06-0.5 microm diameter), and since larger particles (0.5-2.5 microm) have lower penetration rates and higher deposition rates, these estimates are likely to be higher than the true infiltration factors for PM2.5 as a whole. In summer when air conditioners were in use, the sulfur-based infiltration factor was at its lowest (averaging 0.50) for most homes, whereas the average Finf for the other three seasons was 0.62-0.63. Using the daily estimated infiltration factor for each house, we calculated the contribution of outdoor PM2.5 to indoor air concentrations. The indoor-generated contributions to indoor PM2.5 had a wider range (0-33 microg/m3) than the outdoor contributions (5-22 microg/m3). However, outdoor contributions exceeded the indoor-generated contributions in 27 of 36 homes. A second goal of the study was to determine the contribution of outdoor particles to personal exposure. This is determined by the "outdoor exposure factor" Fpex, the fraction of outdoor PM2.5 contributing to personal exposure. As with Finf, we estimated Fpex by the personal/outdoor sulfur ratios. The estimates ranged from 0.33 to 0.77 with a median value of 0.53. Outdoor air particles were less important for personal exposures than for indoor concentrations, with the median outdoor contribution to personal exposure just 49%. We regressed the outdoor contributions to personal exposures on measured outdoor PM2.5 at the central site. The regressions had R2 values ranging from 0.19 to 0.88 (median = 0.73). These values provide an indication of the extent of misclassification error in epidemiological estimates of the effect of outdoor particles on health. PMID- 15819229 TI - Comparison between back-trajectory based modeling and Lagrangian backward dispersion modeling for locating sources of reactive gaseous mercury. AB - Reactive gaseous mercury (RGM) was measured using an annular denuder coated with potassium chloride at three rural sites (Potsdam, Stockton, and Sterling) in New York State from April 2002 to April 2003. Concentrations of RGM ranged from 0.1 to 84.6 pg m(-3) with large spatial and temporal variation. Potential source contribution function (PSCF), a common receptor modeling tool, was used with these measurements, and source-receptor relationships were calculated using back dispersion and deposition as well as back-trajectories. Modeling results were compared with the RGM emissions inventory, and Spearman rank-order correlation coefficients were calculated. PSCF results incorporating backward dispersion and deposition were better correlated with the emissions inventory than PSCF based on back-trajectories alone. This difference was determined to be mainly due to the inclusion of dispersion rather than deposition. The main sources of RGM were suggested to be coal-fired power plants in New York and Pennsylvania, the large copper smelter in Quebec, and the taconite mining areas around the Great Lakes. PMID- 15819230 TI - Calculations of incremental secondary organic aerosol reactivity. AB - The incremental secondary organic aerosol reactivity (ISOAR) of a species j is defined as the relative incremental change in secondary organic aerosol (SOA) formed per relative incremental change in the amount of species jemitted. The California Institute of Technology three-dimensional air quality model is used in conjunction with the Caltech Atmospheric Chemistry Mechanism (CACM) and the Model to Predict the Multiphase Partitioning of Organics to calculate spatially and temporally averaged ISOAR values for the South Coast Air Basin of California (SoCAB). The base case SOA concentrations are derived for September 9, 1993. The South Coast Air Quality Management District of California provided emission and meteorological data. ISOAR values are calculated for the lumped surrogate compounds considered by CACM: isoprene, low-yield monoterpenes, high-yield monoterpenes, high-yield aromatics, etc. This work presents basin-wide ISOAR values determined through regression analysis. In addition, ISOAR values are reported at individual locations within the SoCAB. Modeled data are compared with ISOAR values calculated using smog chamber data. Results indicate that long-chain alkanes present the highest ISOAR. On the other hand, short-chain organics present the lowest ISOAR. PMID- 15819232 TI - Structure of the toxaphene compound 2,5-endo,6-exo,8,9,9,10,10-octachlorobornene 2: a temperature-dependent formation of two rotamers. AB - The irradiation of 2,2,3-exo,5-endo,6-exo,8,9,9,10,10-decachlorobornane in n hexane at 254 nm leads to a spontaneous Cl2 elimination as the major reaction pathway. This results finally in the main product 2,5-endo,6-exo,8,9,9,10,10 octachlorobornene-2, of which the structure could be elucidated with the help of X-ray, 1H and 13C NMR, IR, and MS. Temperature-dependent 1H NMR spectroscopic investigations have shown that the -CHCl2 groups located at C1 and C7 are able to rotate slowly under normal circumstances. If such measurements, however, are exerted at low temperatures (-10 to -60 degrees C), so can be seen that two rotamers are formed due to the hindrance of the free rotation about the bonds C1 C10, C7-C8, and C7-C9, which for the first time could be revealed for a toxaphene compound. Furthermore, as all 1H NMR chlorobornane spectra known so far show only sharp and clear signals, it can be assumed that chlorobornane compounds as main toxaphene components have fixed bonds, which requires to indicate chlorine atoms within the tentacles such as "a", "b", and "c" for characterizing the correct position. Those fixed tentacles are probably the reason that many toxaphene congeners remain very stable in environmental compartments, and particularly the biotic and abiotic transformation may strongly be hindered by the inflexibility of the tentacles. PMID- 15819231 TI - Comparison of centrifugation and filtration techniques for the size fractionation of colloidal material in soil suspensions using sedimentation field-flow fractionation. AB - Sedimentation field-flow fractionation (SdFFF) with UV detection is used to systematically investigate the effect of traditional membrane filtration and centrifugation procedures on the isolation of specific size fractions from soil suspensions. Both procedures were used to isolate the nominal <0.45 and <0.2 microm fractions from a clay soil suspension. Results showed that the membrane filtration approach seriously underestimated the total mass of particulate matter present as compared to the centrifugation approach. This has serious implications forthe interpretation of results for "colloidal" and "soluble" fractions from soil suspensions and other environmental matrices obtained using the standard membrane approach. The results also show that sedimentation FFF has great potential as a robust and relatively mild technology for studying size distributions in the "colloidal" range for soil suspensions and other aquatic matrices. PMID- 15819233 TI - Validation and interpretation of CALUX as a tool for the estimation of dioxin like activity in marine biological matrixes. AB - Among the different analytical tools proposed as an alternative to the very expensive gas chromatography high-resolution mass spectrometry (GC-HRMS) analyses of polychlorodibenzo-p-dioxin and polychlorodibenzofurans, Chemically Activated LUciferase gene eXpression (CALUX) in vitro cell bioassay is very promising. It allows the analyses of a high number of samples since it is relatively fast, inexpensive, and sensitive. However, this technique is not yet widely applied for screening or environmental monitoring. The main reasons are probably the lack of validation and the difficulty in interpreting the global biological response of the bioassay. In this paper, the strict quality control criteria set up for the validation of CALUX are described. The validation has shown good repeatability (relative standard deviation (RSD) = 9%) and good within-lab reproducibility (RSD = 15%) of the results. The quantification limit, in the conditions applied in this paper, is 1.25 pg CALUX-TEQ/g fat. Comparison of CALUX and GC-HRMS analysis was made forvarious marine matrixes (fishes, mussels, starfishes, sea birds, and marine mammals). Good correlations are usually observed, but there are systematic differences between the results. Attempts are made to identify the origin of the discrepancy between the two methods. PMID- 15819234 TI - Modified isotope pairing technique to study N transformations in polluted aquatic systems: theory. AB - Denitrification of nitrate in sediments of polluted aquatic systems has the potential to release considerable nitrogen amounts into the atmosphere in the form of dinitrogen (N2) and/or nitrous oxide (N2O). Nitrate may diffuse into the anoxic sediment layer either from the oxic sediment layer (after being produced there by nitrifiers) or directly from the overlying water. Currently, the most common technique that explicitly distinguishes between the two main nitrate sources is the Isotope Pairing Technique (IPT). 15N-labeled nitrate is added to the water column, and formation of 15N labeled N2 molecules is subsequently monitored. The main shortcoming of the IPT is that the formation of N20 is ignored, thus resulting in an underestimation of sediment denitrification. Another limitation is the inability to account for a possible influx of nitrate through the hyporheic zone (e.g., nitrate-polluted groundwater) into the anoxic sediment layer. We have further developed and analyzed the theoretical basis of the original IPT. The two important factors that we took into account are the isotopic composition of N20 and the input of an external source of nitrate. We also examined the option of adding 15N-labeled ammonium to the water column to specifically adjustthe technique to field studies. The presented modified technique allows us to (i) improve the estimation of sediment denitrification capacity, (ii) gain an insight into the N20 formation mechanism(s) and fluxes, and (iii) assess inputs of nitrate-polluted water through the hyporheic zone. PMID- 15819235 TI - Cu and Cd effects on the earthworm Lumbricus rubellus in the laboratory: multivariate statistical analysis of relationships between exposure, biomarkers, and ecologically relevant parameters. AB - This study sets out to examine the potential of a suite of novel molecular biomarkers as early warning indicators of environmental state and damage. Transcriptional responses of four genes, metallothionein 1 and 2, amine oxidase, and the lysosomal associated glycoprotein, were measured in the earthworm Lumbricus rubellus exposed to increasing concentrations of cadmium and copper in OECD soil. These responses were compared to metal body concentrations and lifecycle parameters: survival, cocoon production, and growth. Adverse physiological effects were observed at concentrations 1/3rd to 1/10th those of the artificial soil LC50. Multivariate statistics, principal component analysis (PCA), was used to investigate the correlations between the different variables. Three key components were derived explaining 77.6% of the variance, with component 1 contributing 32.4%, component 2 contributing 26.7%, and component 3 contributing 18.5%. These components were interpreted in terms of population health, pollutant exposure, and detoxification pathways, respectively. It is proposed that the use of such a suite of biomarkers could serve as indicators of the "health" of the soil environment and provide early warning signals of potential danger to the biota or as a means of monitoring soil remediation. PMID- 15819236 TI - Positron annihilation spectroscopic evidence to demonstrate the flux-enhancement mechanism in morphology-controlled thin-film-composite (TFC) membrane. AB - In this study, positron annihilation lifetime spectroscopy (PALS) is applied to explain the flux-enhancement mechanism in thin-film-composite (TFC) membranes prepared by using dimethyl sulfoxide (DMSO) as an additive in the interfacial polymerization. The TFC membranes show a large increase in water flux, up to 5 fold, compared to nonadditive membrane. Atomic force microscopy (AFM) shows that surface roughness and surface area increase when DMSO in the aqueous phase solution phase works to increase miscibility of the aqueous and the organic phase by reducing the solubility difference of two immiscible solutions. X-ray photoelectron spectroscopy (XPS) reveals the variation of the chemical compositions to the extent that there is a considerable increase in the cross linked amide linkages of the flux-enhanced TFC membranes. The effects of these structural changes on the molecular-size free volume properties are evaluated by PALS studies. The PALS results are the first to experimentally show that the thin films of cross-linked aromatic polyamide RO membranes are composed of two types of pores having radii of about 2.1-2.4 A from tau3, network pore, and 3.5-4.5 A from tau4, aggregate pore. The increase in the size and number of network pores by means of DMSO addition during interfacial polymerization enhances the water flux notably. The size of aggregate pores also increases and may contribute to enhance water flux, although their number inevitably decreases as the number of network pores becomes increased. Details on the correlations between RO performances and o-Ps lifetime parameters are clearly described based on the pore flow model of reverse osmosis developed by Sourirajan et al. PMID- 15819237 TI - Field trial of transgenic Indian mustard plants shows enhanced phytoremediation of selenium-contaminated sediment. AB - Three transgenic Indian mustard [Brassica juncea (L.) Czern.] lines were tested under field conditions for their ability to remove selenium (Se) from Se- and boron-contaminated saline sediment. The transgenic lines overexpressed genes encoding the enzymes adenosine triphosphate sulfurylase (APS), gamma-glutamyl cysteine synthetase (ECS), and glutathione synthetase (GS), respectively. The APS, ECS, and GS transgenic plants accumulated 4.3, 2.8, and 2.3-fold more Se in their leaves than wild type, respectively (P < 0.05). GS plants significantly tolerated the contaminated soil better than wild type, attaining an aboveground biomass/area almost 80% of that of GS plants grown on clean soil, compared to 50% for wild type plants. This is the first report showing that plants genetically engineered for phytoremediation can perform successfully under field conditions. PMID- 15819238 TI - Pilot-scale demonstration of surfactant-enhanced PCE solubilization at the Bachman Road site. 1. Site characterization and test design. AB - A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted to recover dense nonaqueous phase liquid (DNAPL) tetrachloroethene (PCE) from a sandy glacial outwash aquifer underlying a former dry cleaning facility at the Bachman Road site in Oscoda, MI. Part one of this two-part paper describes site characterization efforts and a comprehensive approach to SEAR test design, effectively integrating laboratory and modeling studies. Aquifer coring and drive point sampling suggested the presence of PCE-DNAPL in a zone beneath an occupied building. A narrow PCE plume emanating from the vicinity of this building discharges into Lake Huron. The shallow unconfined aquifer, characterized by relatively homogeneous fine-medium sand deposits, an underlying clay layer, and the absence of significant PCE transformation products, was judged suitable for the demonstration of SEAR. Tween 80 was selected for application based upon its favorable solubilization performance in batch and two dimensional sand tank treatability studies, biodegradation potential, and regulatory acceptance. Three-dimensional flow and transport models were employed to develop a robust design for surfactant delivery and recovery. Physical and fiscal constraints led to an unusual hydraulic design, in which surfactant was flushed across the regional groundwater gradient, facilitating the delivery of concentrations of Tween 80 exceeding 1% (wt) throughout the treatment zone. The potential influence of small-scale heterogeneity on PCE-DNAPL distribution and SEAR performance was assessed through numerical simulations incorporating geostatistical permeability fields based upon available core data. For the examined conditions simulated PCE recoveries ranged from 94to 99%. The effluent treatment system design consisted of low-profile air strippers coupled with carbon adsorption to trap off-gas PCE and discharge of treated aqueous effluent to a local wastewater treatment plant. The systematic and comprehensive design methodology described herein may serve as a template for application at other DNAPL sites. PMID- 15819239 TI - Pilot-scale demonstration of surfactant-enhanced PCE solubilization at the Bachman Road site. 2. System operation and evaluation. AB - A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted during the summer of 2000 at the Bachman Road site in Oscoda, MI. Part two of this two-part paper describes results from partitioning and nonpartitioning tracer tests, SEAR operations, and post-treatment monitoring. For this field test, 68 400 L of an aqueous solution of 6% (wt) Tween 80 were injected to recover tetrachloroethene-nonaqueous phase liquid (PCE-DNAPL) from a shallow, unconfined aquifer. Results of a nonreactive tracer test, conducted prior to introducing the surfactant solution, demonstrate target zone sweep and hydraulic control, confirming design-phase model predictions. Partitioning tracer test results suggest PCE-DNAPL saturations of up to 0.74% within the pilot-scale treatment zone, consistent with soil core data collected during site characterization. Analyses of effluent samples taken from the extraction well during SEAR operations indicate that a total of 19 L of PCE and 95% of the injected surfactant were recovered. Post-treatment monitoring indicated that PCE concentrations at many locations within the treated zone were reduced by as much as 2 orders of magnitude from pre-SEAR levels and had not rebounded 450 days after SEAR operations ceased. Pilot-scale costs ($365 900) compare favorably with design-phase cost estimates, with approximately 10% of total costs attributable to the intense sampling density and frequency. Results of this pilot-scale test indicate that careful design and implementation of SEAR can result in effective DNAPL mass removal and a substantial reduction in aqueous concentrations within the treated source zone under favorable geologic conditions PMID- 15819240 TI - Sulfide-oxidizing activity and bacterial community structure in a fluidized bed reactor from a zero-discharge mariculture system. AB - In the present work we describe a comprehensive analysis of sulfide oxidation in a fluidized bed reactor (FBR) from an environmentally sustainable, zero-discharge mariculture system. The FBR received oxygen-depleted effluent from a digestion basin (DB) that is responsible for gasification of organic matter and nitrogen. The FBR is a crucial component in this recirculating system because it safeguards the fish from the toxic sulfide produced in the DB. Microscale sulfide oxidation potential and bacterial community composition within FBR biofilms were correlated to biofilter performance by integrating bulk chemical, microsensor (O2, pH, and H2S), and molecular microbial community analyses. The FBR consistently oxidized sulfide during two years of continuous operation, with an estimated average sulfide removal rate of 1.3 g of sulfide-S L(FBR)(-1) d(-1). Maximum sulfide oxidation rates within the FBR biofilms were 0.36 and 0.21 mg of sulfide-S cm(-3) h(-1) in the oxic and anoxic layers, respectively, indicating that both oxygen and nitrate serve as electron acceptors for sulfide oxidation. The estimated anoxic sulfide removal rate, as extrapolated from bench scale, autotrophic, nitrate-amended experiments, was 0.7 g of sulfide-S L(FBR)(-1) d(-1), which is approximately 50% of the total estimated sulfide removal in the FBR. Community composition analyses using denaturing gradient gel electrophoresis (DGGE) of bacterial 16S rRNA gene fragments from FBR samples taken at six-month intervals revealed several sequences that were closely affiliated with sulfide-oxidizing bacteria. These included the denitrifying, sulfide-oxidizing bacteria Thiomicrospira denitrificans, members of the filamentous Thiothrix genus, and sulfide-oxidizing symbionts from the Gammaproteobacteria. In addition, marine Alphaproteobacteria and Bacteroidetes species were present in all of the DGGE profiles examined. DGGE analyses showed significant shifts in the bacterial community composition between profiles over two years of sampling, indicating the presence of a diverse and dynamic microbial community within the functionally stable FBR. The FBR's combined capacity for both oxic and anoxic sulfide oxidation, as indicated by bulk chemical, microsensor, and molecular microbial analyses, gives it significant functional elasticity, which is crucial for proper performance in the dynamic environment of this mariculture system. PMID- 15819241 TI - Kinetics and modeling of the Fe(III)/H2O2 system in the presence of sulfate in acidic aqueous solutions. AB - This work examined the effect of sulfate ions on the rate of decomposition of H2O2 by Fe(III) in homogeneous aqueous solutions. Experiments were carried out at 25 degrees C, pH < or = 3 and the concentrations of sulfate ranged from 0 to 200 mM ([Fe(III)]0 = 0.2 or 1 mM, [H2O2]0 = 10 or 50 mM). The spectrophometric study shows that addition of sulfate decreased the formation of iron(III)-peroxo complexes and that H2O2 does not form complexes with iron(III)-sulfato complexes. The rates of decomposition of H2O2 markedly decreased in the presence of sulfate. The measured rates were accurately predicted by a kinetic model based on reactions previously validated in NaClO4/HClO4 solutions and on additional reactions involving sulfate ions and sulfate radicals. At a fixed pH, the pseudo first-order rate constants were found to decrease linearly with the molar fraction of Fe(II) complexed with sulfate. The model was also able to predict the rate of oxidation of a probe compound (atrazine) by Fe(III)/H2O2. Computer simulations indicate that the decrease of the rate of oxidation of organic solutes by Fe(III)/H2O2 can be mainly attributed to the complexation of Fe(III) by sulfate ions, while sulfate radicals play a minor role on the overall reaction rates. PMID- 15819242 TI - Dual electrodes oxidation of dye wastewater with gas diffusion cathode. AB - The high energy cost of an electrochemical method is the fatal drawback that hinders its large scale application in wastewater treatment. In traditional single-chamber electrolysis cell, only direct oxidation at an anode exists. Although a small amount of hydrogen peroxide is produced at the cathode by reduction, it is transferred to the anode and destroyed there without adding much benefit to organic decomposition. A two-chamber electrolytic cell, connected with an electrolyte bridge, was developed in this work. In this new reactor, direct oxidation at anode and indirect oxidation by hydrogen peroxide at cathode can occur simultaneously. Therefore "dual electrodes oxidation" in one electrochemical reactor was achieved successfully. Compared to a traditional one cell reactor, this reactor cuts the energy cost by 50%, and thus might lead to reconsideration of the electrochemical role in wastewater treatment. A Pt/C gas diffusion electrode (GDE) is fabricated and used as a cathode fed with oxygen containing gases to produce hydrogen peroxide. When purified air diffuses through the active layer on the GDE, oxygen is reduced to hydrogen peroxide with a high yield to decompose organics. It has been found that the direct oxidation process at an anodic zone is slightly affected by factors such as pH variation, Fe(II) existence and aeration, while indirect oxidation at the cathodic zone is strongly affected. Dye used as a model pollutant was oxidized into small organic acids in both anode and cathode regions in this electrolytic reactor. GC-MS and IR spectrum were employed to analyze the intermediates formed during the degradation. Twenty intermediates have been detected, including 14 esters, 3 acids and 3 compounds with NO2 or N-OH groups. Thereafter, the degradation pathways of dye Acid Red B are proposed. PMID- 15819243 TI - Photocatalytic oxidation of arsenic(III): evidence of hydroxyl radicals. AB - Arsenic contamination has been found in the groundwater of several countries. Photocatalysis can rapidly oxidize arsenite (As(III)) to less labile and less toxic arsenate (As(V)), which then can be removed by adsorption onto photocatalyst surfaces. This study investigates the photocatalytic oxidation of As(III) to As(V) as a function of As(III) concentration, pH, catalyst loading, light intensity, dissolved oxygen concentration, type of TiO2 surfaces, and ferric ions to understand the kinetics and the mechanism of As(III) oxidation in the UV/TiO2 system. Photocatalytic oxidation of As(III) to As(V) takes place in minutes and follows zero-order kinetics. Benzoic acid (BA) was used as a hydroxyl radical (.OH) scavenger to provide evidence for the .OH as the main oxidant for oxidation of As(III). The .OH radical was independently generated by nitrate photolysis, and kinetics of As(III) oxidation by the .OH radical was determined. Formation of salicylic acid (SA) from the oxidation of BA by .OH also demonstrates the involvement of .OH in the mechanism of As(III) oxidation. The effect of Fe(III) on As(III) oxidation at different pH values with and without TiO2 under UV light was examined. The results suggest that .OH is the dominant oxidant for As(III) oxidation. Two commercially available TiO2 suspensions, Degussa P25 and Hombikat UV100, were tested for the removal of arsenic through oxidation of As(III) to As(V) followed by adsorption of As(V) onto TiO2 surfaces. Results showed that complete removal of arsenic below the World Health Organization drinking water limit of 10 microg/L could be achieved. PMID- 15819244 TI - Catalytic ozonation of gaseous reduced sulfur compounds using wood fly ash. AB - The feasibility of reusing wood ash as an inexpensive catalyst in a catalytic ozonation process has been demonstrated. Catalytic ozonation was demonstrated to oxidize H2S, methanethiol (MT), dimethyl sulfide (DMS), and dimethyl disulfide (DMDS) at low temperatures (23-25 degrees C). The process oxidized 25-50% of an inlet MT stream at 70 ppmv without the formation of DMDS (contrary to ash plus oxygen in air), oxidized 90-95% of an 85 ppmv stream of DMS, and oxidized 50% of a 100 ppmv DMDS stream using 2 g of wood ash at a space velocity of 720 h(-1) using ozone concentrations ranging from 100 to 300 ppmv. Similarly, 60-70% conversion of a 70 ppmv H2S stream was achieved with 2 g of ash in 1.1 s without catalytic deactivation (approximately 44 h). The overall oxidation rate of H2S, DMS, and DMDS increased with increasing ozone concentration contrary to the oxidation rate of MT, which was independent of ozone concentration. Dimethyl sulfoxide and dimethyl sulfone were identified as the primary end products of DMS oxidation, and SO2 was the end product of H2S and MT oxidation. PMID- 15819245 TI - Mineralization of aromatics in water by sunlight-assisted electro-fenton technology in a pilot reactor. AB - The viability of the degradation of aqueous solutions of aniline, nitrobenzene, and 4-chlorophenol by the so-called Photoelectro-Fenton process in a pilot reactor is reported. The Electro-Fenton process stage, based on the flow of oxygen through a gas diffusion cathode to produce H2O2, allows mineralization of about a half of the TOC content in ca. 1 h, because anodic oxidation is coupled with Fenton reaction in the presence of Fe2+ catalyst. An intensity of 20 A was applied in a flow cell comprising a Ti/Pt anode and a carbon-PTFE cathode. After electrolysis, samples of the effluent were exposed to sunlight, and almost complete mineralization was reached after ca. 50 min. Effect of parameters such as electrolysis time, pH, initial concentration, and solar or UVA irradiation on the process efficiencies and the running costs are studied. The Photoelectro Fenton process using sunlight effectively diminishes these costs. The analogous behavior of the three aromatics suggests similar degradation pathways, regardless of the different groups attached to the ring. PMID- 15819246 TI - In situ burning of oil in coastal marshes. 1. Vegetation recovery and soil temperature as a function of water depth, oil type, and marsh type. AB - In-situ burning of oiled wetlands potentially provides a cleanup technique that is generally consistent with present wetland management procedures. The effects of water depth (+10, +2, and -2 cm), oil type (crude and diesel), and oil penetration of sediment before the burn on the relationship between vegetation recovery and soil temperature for three coastal marsh types were investigated. The water depth over the soil surface during in-situ burning was a key factor controlling marsh plant recovery. Both the 10- and 2-cm water depths were sufficient to protect marsh vegetation from burning impacts, with surface soil temperatures of <35 and 48 degrees C, respectively. Plant survival rate and growth responses at these water depth burns were not significantly different from the unburned control. In contrast, a water table 2 cm below the soil surface during the burn resulted in high soil temperatures, with 90-200 degrees C at 0 0.5 cm soil depth and 55-75 degrees C at 1-2 cm soil depth. The 2-cm soil exposure to fire significantly impeded the post-burn recovery of Spartina alterniflora and Sagittaria lancifolia but did not detrimentally affect the recovery of Spartina patens and Distichlis spicata. Oil type (crude vs diesel) and oil applied to the marsh soil surface (0.5 L x m(-2)) before the burn did not significantly affect plant recovery. Thus, recovery is species-specific when no surface water exists. Even water at the soil surface will most likely protect wetland plants from burning impact. PMID- 15819247 TI - In-situ burning of oil in coastal marshes. 2. Oil spill cleanup efficiency as a function of oil type, marsh type, and water depth. AB - In-situ burning of spilled oil, which receives considerable attention in marine conditions, could be an effective way to cleanup wetland oil spills. An experimental in-situ burn was conducted to study the effects of oil type, marsh type, and water depth on oil chemistry and oil removal efficiency from the water surface and sediment. In-situ burning decreased the totaltargeted alkanes and total targeted polycyclic aromatic hydrocarbons (PAHs) in the burn residues as compared to the pre-burn diesel and crude oils. Removal was even more effective for short-chain alkanes and low ring-number PAHs. Removal efficiencies for alkanes and PAHs were >98% in terms of mass balance although concentrations of some long-chain alkanes and high ring-number PAHs increased in the burn residue as compared to the pre-burn oils. Thus, in-situ burning potentially prevents floating oil from drifting into and contaminating adjacent habitats and penetrating the sediment. In addition, in-situ burning significantly removed diesel oil that had penetrated the sediment for all water depths. Furthermore, in situ burning at a water depth 2 cm below the soil surface significantly removed crude oil that had penetrated the sediment. As a result, in-situ burning may reduce the long-term impacts of oil on benthic organisms. PMID- 15819248 TI - Emission factors for carbonaceous particles and polycyclic aromatic hydrocarbons from residential coal combustion in China. AB - Emission factors of carbonaceous particles, including black carbon (BC) and organic carbon (OC), and polycyclic aromatic hydrocarbons (PAHs) were determined for five coals, which ranged in maturity from sub-bituminous to anthracite. They were burned in the form of honeycomb briquettes in a residential coalstove, one of the most common fuel/stove combinations in China. Smoke samples were taken through dilution sampling equipment, with a high volume sampler that could simultaneously collect emissions in both particulate and gaseous phases, and a cascade impactor that could segregate particles into six fractions. Particulate BC and OC were analyzed by a thermal-optical method, and PAHs in emissions of both phases were analyzed by GC-MS. Burning of bituminous coals produced the highest emission factors of particulate matter (12.91 g/kg), BC (0.28 g/kg), OC (7.82 g/kg), and 20 PAHs (210.6 mg/kg) on the basis of burned dry ash-free (daf) coal, while the anthracite honeycomb-briquette was the cleanest household coal fuel. The size-segregated results show that more than 94% of the particles were submicron, and calculated mass median aerodynamic diameters (MMAD) of all particles were under 0.3 microm. Based on the coal consumption in the residential sector of China, 290.24 Gg (gigagrams) of particulate matter, 5.36 Gg of BC, 170.33 Gg of OC, and 4.72 Gg of 20 PAHs mass were emitted annually from household honeycomb-briquette burning during 2000. Anthracite coal should be selected preferentially and more advanced burning conditions should be applied in domestic combustion, from the viewpoint of both climate change and adverse health effects. PMID- 15819249 TI - Major anion effects on the kinetics and reactivity of granular iron in glass encased magnet batch reactor experiments. AB - The relative effects of sulfate (SO4(2-)), chloride (Cl-), nitrate (NO3-), and bicarbonate (HCO3-) (8 mM ionic strength solutions, adjusted to pH 10) on the reactivity of Master Builders iron was investigated using a low-abrasion batch reactor with a glass-encased magnet (GEM). Reactivity of the granular iron surface was assessed by measuring the reduction rate of 4-chloronitrobenzene (4ClNB) as a function of initial 4CINB concentration and anion type. Relative to a similarly prepared perchlorate (ClO4-) solution, in which perchlorate was assumed not to interact with the iron surface, nitrate and bicarbonate inhibited the reduction of the probe compound (4ClNB). Chloride and sulfate enhanced reactivity. Thus, the anions were ranked SO4(2-) > Cl- > or = ClO4- > NO3- > HCO3 (from most enhanced to most inhibited) in their influence on granular iron reactivity toward 4ClNB. Kinetic studies of 4CINB were conducted under conditions that caused the iron surface to saturate with the reacting compound (saturation kinetic studies). These experiments, conducted in the various anion solutions indicated above, showed that the gains in reactivity that occurred in the presence of Cl- and SO4(2-) were due to either increased surface reactivity or sorption capacity. The losses in reactivity that occurred in the presence of NO3- were due to decreases in one or both of these same two factors. However, reactivity declines in the presence of CO3(2-) appear to have been due, in large part, to a reduced affinity of 4ClNB for the iron surface. PMID- 15819250 TI - Electroreduction of Cr(VI) to Cr(III) on reticulated vitreous carbon electrodes in a parallel-plate reactor with recirculation. AB - The reduction of Cr(VI) to Cr(III) is achieved in a flow-by, parallel-plate reactor equipped with reticulated vitreous carbon (RVC) electrodes;this reduction can be accomplished by the application of relatively small potentials. Treatment of synthetic samples and field samples (from an electrodeposition plant) results in final Cr(VI) concentrations of 0.1 mg/L (i.e., the detection limit of the UV vis characterization technique used here) in 25 and 43 min, respectively. Such concentrations comply with typical environmental legislation for wastewaters that regulate industrial effluents (at presenttime = 0.5 mg/L for discharges). The results show the influence of the applied potential, pH, electrode porosity, volumetric flow, and solution concentration on the Cr(VI) reduction percentage and on the required electrolysis time. Values for the mass transfer coefficient and current efficiencies are also obtained. Although current efficiencies are not high, the fast kinetics observed make this proposed treatment an appealing alternative. The lower current efficiency obtained in the case of a field sample is attributed to electrochemical activation of impurities. The required times for the reduction of Cr(VI) are significantly lower than those reported elsewhere. PMID- 15819251 TI - TiO2-photocatalyzed As(II) oxidation in aqueous suspensions: reaction kinetics and effects of adsorption. AB - Oxidation of arsenite, As(III), to arsenate, As(V), is required for the efficient removal of arsenic by many water treatment technologies. The photocatalyzed oxidation of As(III) on titanium dioxide, TiO2, offers an environmentally benign method for this unit operation. In this study, we explore the efficacy and mechanism of TiO2-photocatalyzed As(III) oxidation at circumneutral pH and over a range of As(III) concentrations approaching those typically encountered in water treatment systems. We focus on the effect of As adsorption on observed rates of photooxidation. Adsorption (in the dark) of both As(III) and As(V) on Degussa P25 TiO2 was examined at pH 6.3 over a range in dissolved arsenic concentrations, [As]diss, of 0.10-89 microM and 0.2 or 0.05 g L(-1) TiO2 for As(III) and As(V), respectively. Adsorption isotherms generally followed the Langmuir-Hinshelwood model with As(III) exhibiting an adsorption maxima of 32 micromol g(-1). As(V) adsorption did not reach a plateau under the experimental conditions examined; the maximum adsorbed concentration observed was 130 micromol g(-1). The extent of As(III) and As(V) adsorption observed at the beginning and end of the kinetic studies was consistent with that observed in the adsorption isotherms. Kinetic studies were performed in batch systems at pH 6.3 with 0.8-42 microM As(III) and 0.05 g L(-1) TiO2; complete oxidation of As(III) was observed within 10-60 min of irradiation at 365 nm. The observed effect of As(III) concentration on reaction kinetics was consistent with surface saturation at higher concentrations. Addition of phosphate at 0.5-10 microM had little effect on either As(III) sorption or its photooxidation rate but did inhibit adsorption of the product As(V). The selective use of hydroxyl radical quenchers and superoxide dismutase demonstrated that superoxide, O2-, plays a major role in the oxidation of As(III) to As(V). PMID- 15819252 TI - Influence of diesel engine combustion parameters on primary soot particle diameter. AB - Effects of engine operating parameters and fuel composition on both primary soot particle diameter and particle number size distribution in the exhaust of a direct-injected heavy-duty diesel engine were studied in detail. An electrostatic sampler was developed to deposit particles directly on transmission electron microscopy (TEM) grids. Using TEM, the projected area equivalent diameter of primary soot particles was determined. A scanning mobility particle sizer (SMPS) was used for the measurement of the particle number size distribution. Variations in the main engine operating parameters (fuel injection system, air management, and fuel properties) were made to investigate soot formation and oxidation processes. Primary soot particle diameters determined by TEM measurements ranged from 17.5 to 32.5 nm for the diesel fuel and from 24.1 to 27.2 nm for the water diesel emulsion fuel depending on the engine settings. For constant fuel energy flow rate, the primary particle size from the water-diesel emulsion fuel was slightly larger than that from the diesel fuel. A reduction in primary soot particle diameter was registered when increasing the fuel injection pressure (IP) or advancing the start of injection (SOI). Larger primary soot particle diameters were measured while the engine was operating with exhaust gas recirculation (EGR). Heat release rate analysis of the combustion process revealed that the primary soot particle diameter decreased when the maximum flame temperature increased for the diesel fuel. PMID- 15819253 TI - Controlling hydrothermal reaction pathways to improve acetic acid production from carbohydrate biomass. AB - A two-step hydrothermal process to improve the production of acetic acid was discussed. The first step was to accelerate the formation of 5-hydroxymethyl-2 furaldehyde (HMF), 2-furaldehyde (2-FA), and lactic acid (LA), and the second step was to further convert the furans (HMF, 2-FA) and LA produced in the first step to acetic acid by oxidation with newly supplied oxygen. The acetic acid obtained by the two-step process had not only a high yield but also better purity. The contribution of two pathways via furans and LA in the two-step process to convert carbohydrates into acetic acid was roughly estimated as 85 90%, and the ratio of the contributions of furans and LA to yield acetic acid was estimated as 2:1. The fact that WO of carbohydrates is not capable of producing a large amount of acetic acid, while the two-step process can enhance the acetic acid yield, can be explained because formic acid is a basic product of direct oxidation of carbohydrate, and acetic acid in WO of carbohydrates may come from the oxidation of dehydration products of aldose. PMID- 15819254 TI - Emissions and energy efficiency assessment of baseload wind energy systems. AB - The combination of wind energy generation and energy storage can produce a source of electricity that is functionally equivalent to a baseload coal or nuclear power plant. A model was developed to assess the technical and environmental performance of baseload wind energy systems using compressed air energy storage. The analysis examined several systems that could be operated in the midwestern United States under a variety of operating conditions. The systems can produce substantially more energy than is required from fossil or other primary sources to construct and operate them. By operation at a capacity factor of 80%, each evaluated system achieves an effective primary energy efficiency of at least five times greater than the most efficient fossil combustion technology, with greenhouse gas emission rates less than 20% of the least emitting fossil technology currently available. Life-cycle emission rates of NOx and SO2 are also significantly lower than fossil-based systems. PMID- 15819255 TI - Assessing quantities and disposal routes for household hazardous products in the United Kingdom. AB - The disposal of household products containing hazardous substances (household hazardous wastes; HHW) is of concern due to possible health and environmental effects as a consequence of environmental pollution. The potential risks of disposal are proportional to the amounts of products used and waste generated, but much of the data relating to quantities are old, inconsistent, or nonexistent. Hence, full-scale risk assessment is not yet feasible. This pilot study was aimed at an initial assessment of the amounts of hazardous products used or stored within the household and potential disposal routes. Representatives of 400 households from southeast England were interviewed about socio-demographic factors, perception of the risks associated with the use and disposal of hazardous waste generated in households, quantities of particular products currently in use or stored within the household, and times and methods of disposal of such products. The estimates of quantities obtained were compared with sales figures and waste estimates to improve understanding of product flow through to the HHW stream. The disposal routes investigated demonstrated that most householders claim to use the entire product priorto disposal in the general refuse bin. The relationship with socio-demographic factors demonstrated a difference between neighborhood size and length of residence in a household with regard to product quantities possessed and the disposal habits adopted. PMID- 15819256 TI - Both sides of the story. Tackling abuse; professional regulation. PMID- 15819257 TI - Independent living. PMID- 15819258 TI - Spot the fakes. Nureses are being put on worldwide alert over the sale of counterfeit drugs. PMID- 15819259 TI - Pushing for equality. PMID- 15819260 TI - Emotional rescue. The meaning of 'emotional labour'--the new management buzzwords. PMID- 15819261 TI - On call with care. Work-related injuries do not have to mean the end of a nursing career. PMID- 15819262 TI - Safer working. PMID- 15819263 TI - A solid grounding. PMID- 15819264 TI - Why choose nursing? PMID- 15819265 TI - Common psychological challenges for patients with newly acquired disability. AB - A patient's attitude to a newly acquired disability can often be the determining factor in successful rehabilitation. This attitude is determined by his or her perceived ability to manage symptoms, deal with hospital procedures, form and maintain relationships and manage his or her feelings about the disability. Because of this complexity, psychological aspects of patient rehabilitation are often regarded as vague and difficult to negotiate. This article describes seven common psychological challenges that patients experience with a newly acquired disability or during recovery from major illness (Moos and Schaefer 1984). Strategies are outlined to assist patients to come to terms with their disability and to cope more effectively during rehabilitation. PMID- 15819266 TI - Clinical guidance in diabetes: using the Internet. AB - An internet site has been developed to help nurses caring for patients with diabetes. It aims to offer evidence-based guidance and help to nurses who are charged with implementing the National Service Framework for Diabetes (Department of Health (DH) 2001a) to improve care for patients and reduce variations in practice throughout the UK. This article describes the setting up process for the site and gives an overview of its contents. PMID- 15819267 TI - Management and prevention of abnormal scars. AB - This article examines the formation of abnormal scars and describes how the different types can be treated. Psychological issues may result from scarring. The authors discuss how nurses can help patients to come to terms with altered body image. PMID- 15819268 TI - Step by step progress. Focus on forensic mental health nursing. PMID- 15819269 TI - Physicians' claims of retaliatory action assessed by courts. PMID- 15819270 TI - In pursuit of wellness. PMID- 15819271 TI - New robot technology indicates bright future for prosthetics. PMID- 15819272 TI - Above and beyond. PMID- 15819273 TI - Gait recovery. PMID- 15819274 TI - Liquid assets. PMID- 15819275 TI - Intervening care. PMID- 15819276 TI - Navigating the options. PMID- 15819277 TI - Growing confusion. PMID- 15819278 TI - Show me the impact. PMID- 15819279 TI - Cecilia Makiwane--pioneer African nurse. PMID- 15819280 TI - Identifying patients with sleep apnea. PMID- 15819281 TI - Use of ambulatory overnight oximetry to investigate sleep apnea in a general internal medicine practice. AB - OBJECTIVE: To examine how clinical factors and results from ambulatory overnight oximetry related to recommendations for further sleep evaluation in patients with clinically suspected obstructive sleep apnea syndrome. PATIENTS AND METHODS: We reviewed 100 medical records selected randomly from 375 consecutive patients for whom ambulatory overnight oximetry had been ordered by internists and completed between September 1, 2001, and May 1, 2002. We analyzed relationships among clinical Information, oximetry results, resultant recommendations, and patient follow-up data. RESULTS: Only 21 of 100 patients had normal results from ambulatory overnight oximetry; 5 were referred for further sleep consultation. Abnormal results from ambulatory overnight oximetry were seen in 79 patients, but only 51 were referred for further sleep evaluation. Abnormal results from ambulatory overnight oximetry were associated with only a small increased likelihood of referral (likelihood ratio, 2.7; confidence interval, 1.2-6.0). Those with an oxygen desaturation index (number of desaturation events per hour of recording time) of greater than 15 received sleep consultation at a median of 8 days after completion of oximetry, whereas those with an oxygen desaturation index of 6 to 10 were evaluated in a median of 42 days (P=.60). All 17 patients who had minimum oxygen saturation of less than 80% were referred for further evaluation. CONCLUSIONS: Abnormal results from ambulatory overnight oximetry per se may not substantially influence internist referral of patients with clinically suspected sleep apnea for further sleep evaluation. Rather, severity of oximetry abnormalities is used along with other patient-related factors and sleep study accessibility to prioritize the need and urgency of further evaluation. PMID- 15819282 TI - Sarcomeric genotyping in hypertrophic cardiomyopathy. AB - OBJECTIVE: To pool results from studies of patients with hypertrophic cardiomyopathy (HCM) to elucidate important phenotypic differences among genotypes. MATERIAL AND METHODS: Data published from November 1998 through November 2004 were gathered and compared from unrelated study population genotyping studies from the Mayo Clinic (Rochester, Minn), Harvard Medical School (Boston, Mass), France, Germany, Sweden, Finland, and Spain. Standard statistical analysis techniques were used to pool and compare data across genotypes with respect to frequency of mutations, age at diagnosis, and degree of hypertrophy (left ventricular wall thickness). RESULTS: The French study population harbored the highest frequency of mutations (61%), followed by the Mayo Clinic (38%), Harvard Medical School (36%), and Swedish (30%) study populations. For every study population, mutations in myosin binding protein C (MYBPC3) were the most common cause of HCM. Patients with a family history of HCM had mutations more frequently than those without. This pooled analysis revealed no statistically significant differences in left ventricular wall thickness or in mean age at diagnosis across all genotypes. CONCLUSIONS: Differentiation of sarcomeric genotypes, such as MYBPC3-HCM and MYH7-HCM, is not possible on the basis of currently reported phenotypic data. A myriad of genetic and/or environmental modifiers in addition to the primary disease-causing genetic substrate must play an important role in determining a patient's particular phenotype. PMID- 15819283 TI - Evaluation of the comparative efficacy of etoricoxib and ibuprofen for treatment of patients with osteoarthritis: A randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To directly compare the efficacy and safety of etoricoxib, 30 mg once daily, ibuprofen, 800 mg 3 times daily, and placebo for treatment of osteoarthritis (OA) of the hip and knee. PATIENTS AND METHODS: A randomized, double-blind, placebo-controlled trial of patients with OA of the knee or hip was performed between February 2003 and November 2003 in 61 medical centers in the United States. Qualified patients aged 40 to 89 years were randomized to receive placebo, etoricoxib, 30 mg once daily, or ibuprofen, 800 mg 3 times daily, for 12 weeks. Primary efficacy end points Included the Western Ontario and McMaster Universities Osteoarthritis Index pain and physical function subscales and Patient Global Assessment of Disease Status. Response to treatment was assessed by the time-weighted average change from baseline over 12 weeks. RESULTS: In 528 patients, baseline values for the 3 primary end points ranged from 67.78 to 72.60 mm (0-100 mm visual analog scale). Near-maximal efficacy was achieved by week 2 with both active treatments and sustained over the course of the trial. During the 12-week period, least squares mean changes in the primary end points (Western Ontario and McMaster Universities Osteoarthritis Index and Patient Global Assessment of Disease Status subscales) ranged from -16.53 to -13.55 mm, -27.89 to -23.68 mm, and -26.53 to -22.97 mm in the placebo, etoricoxib, and Ibuprofen groups, respectively. Both etoricoxib and ibuprofen were more effective (P<.001) than placebo for all primary end points. Etoricoxib and ibuprofen treatment responses for the primary end points were determined to be comparable with use of prespecified comparability criteria. Results for all other efficacy end points were consistent with responses observed for the primary end points. Etoricoxib and ibuprofen generally were well tolerated. CONCLUSION: For patients with OA, treatment with etoricoxib, 30 mg/d, is well tolerated and provides sustained clinical effectiveness that is superior to placebo and comparable to ibuprofen, 2400 mg/d. PMID- 15819284 TI - Diagnostic yield and utility of neurovascular ultrasonography in the evaluation of patients with syncope. AB - OBJECTIVE: To determine the diagnostic utility of neurovascular ultrasonography (transcranial Doppler and carotid ultrasonography) in patients with syncope. PATIENTS AND METHODS: We retrospectively identified consecutive patients who underwent neurovascular ultrasonography for the diagnosis of syncope or presyncope at an academic hospital in 1997 and 1998. From medical records we abstracted patient demographic and clinical information, results and consequences of testing, and follow-up data for 3 years. RESULTS: A total of 140 patients participated in the study. The median age of the study patients was 74 years (interquartile range, 66-80 years), and 49% were male. Severe extracranial or Intracranial cerebrovascular disease was found on neurovascular ultrasonography in 20 patients (14%; 95% confidence interval [CI], 9.5%-21%). Focal neurologic signs or symptoms or carotid bruits were found in 19 (95%) of 20 patients with positive test results compared with 46 (38%) of 120 patients without severe disease (P<.001). Ultrasonography identified cerebrovascular lesions that may have contributed to the syncopal process in only 2 (1.4%) of 140 patients (95% CI, 0.39%-5.1%), but the lesions were unlikely to have been the primary cause of syncope in either patient. CONCLUSION: In this predominantly stroke-age population, neurovascular ultrasonography had a low yield for diagnosing vascular lesions that contributed to the pathophysiology of syncope. However, in patients with focal signs or symptoms or carotid bruits, it detected incidental lesions that typically require treatment or follow-up. In patients with syncope, neurovascular ultrasonography should be reserved for this subset. The data suggest enhancements to the American College of Physicians guideline for the use of neurovascular ultrasonography in patients with syncope. PMID- 15819285 TI - Risk of anaphylaxis after reexposure to intravenous lepirudin in patients with current or past heparin-induced thrombocytopenia. AB - OBJECTIVE: To retrospectively assess the signs and symptoms indicative of adverse reactions to repeated exposures to lepirudin in patients with heparin-induced thrombocytopenia who received at least 2 courses of lepirudin therapy. PATIENTS AND METHODS: Medical records were retrospectively assessed from adult patients who received at least 2 courses of lepirudin therapy separated by at least 1 week between January 1999 and June 2002 at The Cleveland Clinic, Cleveland, Ohio. We evaluated the list of 289 low-level terms for possible signs and symptoms of anaphylactic reactions In the medical dictionary for regulatory activities as well as patient vital signs to detect manifestations of immediate hypersensitivity reactions. Vital signs from the day before initiation of lepirudin therapy were compared with those from days 1 and 2 after exposure. RESULTS: No cases of anaphylaxis or allergic reaction related to lepirudin administration were identified among 43 adult patients. On day 1 of lepirudin, 10 patients had lower systolic blood pressures (by > or =20 mm Hg) than pre lepirudin values, and 4 patients had systolic blood pressures of less than 100 mm Hg. CONCLUSIONS: Isolated asymptomatic decreases in blood pressure after patient reexposure to lepirudin most likely do not reflect anaphylaxis due to lepirudin. We believe that isolated and uncommon cases of anaphylaxis temporally related to lepirudin exposure should not preclude its use in patients with heparin-induced thrombocytopenia and past lepirudin exposure. PMID- 15819286 TI - Lipid-lowering therapy in patients with peripheral arterial disease: are guidelines being met? AB - OBJECTIVES: To determine the proportion of patients with lower extremity peripheral arterial disease (PAD) who reach recommended low-density lipoprotein cholesterol (LDL-C) levels (<100 mg/dL) and to identify the patient characteristics that are independently associated with attaining the LDL-C goal (<100 mg/dL). PATIENTS AND METHODS: Eligible patients were identified from a roster of patients who had undergone testing at a nonvascular laboratory between September 1, 2001, and January 31, 2002, and were found to have evidence of PAD, defined as an ankle-brachial index of 0.9 or less. We thoroughly reviewed patients' electronic medical records. Backward elimination multivariate logistic regression modeling was used to Identify factors associated with reaching the goal LDL-C level. RESULTS: Among 143 patients with PAD, 105 (73%) met the goal LDL-C level. Lipid-lowering therapy was prescribed for 109 (76%). Lower diastolic blood pressure and lower weight were independently associated with an LDL-C level of less than 100 mg/dL. CONCLUSION: We found higher rates of lipid-lowering therapy in patients with PAD than reported previously. Patients with diabetes mellitus or coronary artery disease were not more likely to meet the goal LDL-C level than those without these comorbidities. Clinical practice may be catching up to clinical guidelines. PMID- 15819287 TI - Rheumatoid pachymeningitis presenting with left-sided weakness. PMID- 15819288 TI - Somatostatin analogues in the treatment of gastroenteropancreatic neuroendocrine tumors. AB - Gastroenteropancreatic neuroendocrine tumors constitute a heterogeneous group of neoplasms that are often associated with typical symptoms due to excessive and uncontrolled release of diverse hormones. Because these tumors are usually slow growing, surgery is the cornerstone of treatment. However, these rare tumors can present with rapid progression that requires aggressive systemic therapy or diffuse metastatic disease not amenable to surgical palliation. For most patients, medical approaches are necessary at some point in the course of their disease, especially since most tumors are at an advanced stage at the time of diagnosis. Most gastroenteropancreatic neuroendocrine tumors express high levels of somatostatin receptors, which are bound by somatostatin or its synthetic analogues. These agents, alone or combined with other therapies, such as Interferon or radioisotopes, are therefore used frequently to control hormone related symptoms and, for some patients, the growth of the disease Itself. This article reviews the evidence for the use of somatostatin analogues in the treatment of gastroenteropancreatic neuroendocrine tumors based on a MEDLINE search of literature published from January 1970 to July 2003. PMID- 15819289 TI - Hypnosis in contemporary medicine. AB - Hypnosis became popular as a treatment for medical conditions in the late 1700s when effective pharmaceutical and surgical treatment options were limited. To determine whether hypnosis has a role in contemporary medicine, relevant trials and a few case reports are reviewed. Despite substantial variation in techniques among the numerous reports, patients treated with hypnosis experienced substantial benefits for many different medical conditions. An expanded role for hypnosis and a larger study of techniques appear to be indicated. PMID- 15819290 TI - Action on obesity: report of a mayo clinic national summit. AB - In May 2004, representatives from local, state, and national public and private organizations met in Rochester, Minn, for the Action on Obesity Summit hosted by Mayo Clinic. The overall goal of this summit was to identify creative and effective strategies to Increase the US population's physical activity and improve nutrition to reverse the increasing prevalence of obesity. Ideas generated from selected abstract presentations and breakout sessions were prioritized and incorporated into an action model (available at www.actiononobesity.org) deemed feasible for implementation into most communities. Highlights of the presentations included a company that reported lower than expected health care expenditures secondary to a work site wellness program, a national initiative to increase physical activity (www.americaonthemove.org), and innovative work site nutritional strategies. The implementation model that emerged contained certain themes. Coordinated action at all levels will be required to substantially impact the increasing prevalence of obesity. Educational messages should be simple, consistent, tailored, and linked to benefits. Healthy food options in vending machines and restaurants and increased opportunities for daily physical activity should be available in schools, work sites, and communities. Legislative and policy changes should promote physical activity and improve nutrition. Support for research should be encouraged and outcome measures for interventions documented. A second Action on Obesity Summit is planned for June 9 and 10, 2005, that will review the progress made in the intervening year and continue to refine the implementation model to help address the obesity epidemic, one of the greatest public health problems facing the United States. PMID- 15819291 TI - 68-year-old man with chronic diarrhea, nausea, and vomiting. PMID- 15819292 TI - Trousseau syndrome and the unknown cancer: use of positron emission tomographic imaging in a patient with a paraneoplastic syndrome. AB - Trousseau syndrome is defined as a migratory thrombophlebitis found typically in patients with an underlying malignancy. Conventional diagnostic testing and imaging can be used to successfully diagnose a primary malignancy in approximately 85% to 95% of patients. However, along with a comprehensive medical history and physical examination, numerous tests are frequently required, including blood tests, tumor markers, chest radiography, upper endoscopy, and computed tomography of the chest, abdomen, and pelvis. We present a case in which positron emission tomographic imaging was important for diagnosing the malignancy underlying Trousseau syndrome. Positron emission tomography may play an important role in the efficient evaluation of such cases. PMID- 15819293 TI - Ischemic stroke associated with use of an ephedra-free dietary supplement containing synephrine. AB - In response to concerns regarding the safety of ephedra-containing dietary supplements, manufacturers have marketed "ephedra-free" products. Many of these contain synephrine, a sympathomimetic amine from the plant Citrus aurantium. Synephrine is structurally similar to ephedrine and has vasoconstrictor properties. We describe a 38-year-old patient with ischemic stroke associated with an ephedra-free dietary supplement containing synephrine and caffeine. The patient presented with memory loss and unsteady gait after taking 1 or 2 capsules per day of a dietary supplement (Stacker 2 Ephedra-Free) for 1 week. He had no notable medical history or major atherosclerotic risk factors and took no other medications. Physical examination showed a mildly ataxic gait and substantial Impairment of both concentration and memory. Computed tomography and magnetic resonance Imaging of the brain showed subacute infarctions in the left thalamus and left cerebellum in the distribution of the vertebrobasilar circulation. Other causes of ischemic stroke were evaluated, and findings were unremarkable; a vasospastic origin was considered most likely. The patient was discharged with nearly complete resolution of symptoms. Synephrine, a sympathomimetic amine related to ephedrine, may be associated with Ischemic stroke. Consumers and clinicians need to be Informed about the potential risks of ephedra-free products. PMID- 15819294 TI - Laparoscopic biopsy and ureterolysis in Erdheim-Chester disease. AB - Erdheim-Chester disease (ECD) is a rare, progressive, non-Langerhans cell histiocytosis that portends a poor prognosis. Misdiagnosis is common until adequate tissue analysis reveals the infiltrative histiocytic process. The disease process affects various organ systems, and effective treatment options are limited. Knowledge of the constellation of signs and symptoms associated with ECD is important to avoid misdiagnosis. We describe a patient who underwent laparoscopic bilateral ureterolysis and laparoscopic biopsy for presumed retroperitoneal fibrosis confirmed previously by percutaneous needle biopsy findings. The final pathologic diagnosis based on laparoscopic biopsy results was ECD. As evidenced by this case, ureterolysis offers little benefit to patients with ECD. PMID- 15819295 TI - Sweet syndrome. PMID- 15819296 TI - Subarachnoid hemorrhage: neurointensive care and aneurysm repair. AB - Aneurysmal subarachnoid hemorrhage (SAH) is often a neurologic catastrophe. Diagnosing SAH can be challenging, and treatment is complex, sophisticated, multidisciplinary, and rarely routine. This review emphasizes treatment in the intensive care unit, surgical and endovascular therapeutic options, and the current state of treatment of major complications such as cerebral vasospasm, acute hydrocephalus, and rebleeding. Outcome assessment in survivors of SAH and controversies in screening of family members are discussed. PMID- 15819297 TI - Likelihood ratio reporting. PMID- 15819298 TI - Likelihood ratio reporting. PMID- 15819299 TI - Antecedent hypertension interacts with adverse ventricular remodeling after myocardial infarction. AB - BACKGROUND: Ventricular remodeling is an independent predictor of left ventricular function in patients with myocardial infarction (MI). Accelerated ventricular remodeling is a probable contributor to the increased mortality observed after MI in hypertensive patients. The purpose of this study is to assess the relationship of antecedent hypertension to ventricular remodeling after MI. METHODS: Seventy-nine patients presenting with a first acute MI (AMI) who were treated and attained Thrombolysis In Myocardial Infarction (TIMI) flow grade 3 with re-perfusion therapy within 12 hours of their symptom onset were enrolled in this study. All of them underwent left ventriculography (LVG) immediately after re-perfusion therapy and again at 6 months after the occurrence of MI. Patients were divided into two groups; hypertensive group and normotensive group. End-diastolic volume index (EDVI), end-systolic volume index (ESVI), and EF were compared between acute phase and 6 months after AMI in each group. RESULTS: The hypertensive group showed a significant increase in both EDVI and ESVI after 6 months, whereas the normotensive group did not. In addition, there was no change in EF in the hypertensive group, whereas EF increased significantly after 6 months in the normotensive group. As a result, percent change in ESVI and EF showed significantly differences between the hypertensive group and normotensive group, whereas percent change in EDVI showed no significant difference between the two groups. CONCLUSIONS: Antecedent hypertension interacts with ventricular remodeling after MI. PMID- 15819300 TI - Impact of HMG-CoA reductase inhibitors for non-treated coronary segments. AB - BACKGROUND: HMG-CoA reductase inhibitors (statin) have been reported to decrease coronary artery events in several angiographic studies. However, the mechanism by which statin achieve this is still unclear. The purpose of this study was to identify the effect of statin on coronary plaque using serial intravascular ultrasound analysis. METHODS AND RESULTS: In this study, 48 patients with 48 lesions were divided into the prescribed group (statin group, n = 22) or the non prescribed group (control group, n = 26) after successful coronary artery stenting. IVUS images were obtained at consecutive 5 mm segments, 5 mm from the proximal stent edge, immediately after stenting and at 6 months follow up. External elastic membrane volume (EEMV), lumen volume (LV) and plaque volume (PV) were measured using Simpson's method. The control group revealed no significant serial change in EEMV, PV, and LV during 6 months. On the other hand, the statin group revealed significant reductions of PV (35.5 +/- 12.7 mm3 vs 30.9 +/- 15.6 mm3, p = 0.001), resulting in increase of LV (47.7 +/- 19.8 mm3 vs 52.5 +/- 22.2 mm3, p = 0.003) without EEMV change (82.8 +/- 21.8 mm3 vs 83.9 +/- 25.7 mm3, p = NS). Although percent EEMV and percent LV changes showed no differences between the two groups, a larger percent PV change was observed in the statin group compared to the control group (control; 5.8 +/- 20.3% vs statin; -20.4 +/- 21.8%, p = 0.02). CONCLUSION: The results of this study suggest that statin administration for 6 months reduces coronary plaque without positive vessel remodeling. PMID- 15819301 TI - The relationship between left ventricular geometric remodeling and cardiac function in essential hypertension. AB - BACKGROUND: Abnormal left ventricular (LV) diastolic relaxation is an early sign of hypertensive heart disease. Whether diastolic dysfunction differs between LV geometric patterns remains controversial. METHODS AND RESULTS: We examined 220 hypertensive patients by echocardiography to assess the relationship between diastolic function and LV geometric pattern. We identified four LV geometric patterns: normal geometry, concentric remodeling, concentric hypertrophy, and eccentric hypertrophy. Diastolic function was evaluated by the E/A ratio, E wave deceleration time (Dct), and isovolumic relaxation time (IRT). The E/A ratio was below one and Dct was prolonged in each group. The duration of IRT in the concentric and eccentric hypertrophy groups was significantly longer than that in normal geometry and concentric remodeling groups (p < 0.05). CONCLUSIONS: Hypertensive patients may be possible to have diastolic dysfunction, regardless of the differences in their structural geometry. The degree of impairment of cardiac diastolic function differed between hypertensive patients with different LV geometric patterns. PMID- 15819302 TI - The impact of IVUS guided bare metal stent implantation for non-small vessel. AB - Previous studies have shown larger target vessel or greater stent area contribute to reduced restenosis rate. Thus, intravascular ultrasound (IVUS)-guided bare metal stent (BMS) implantation for selected lesions might achieve drug-eluting stent-like outcomes. The aim was to examine the long-term outcomes of BMS using IVUS-guided optimization. METHODS: Consecutive 103 coronary artery lesions suitable for IVUS were enrolled. Using IVUS, final stent balloon size selected was 90 percent of media-to-media diameter at the lesion or distal reference. The balloon size was listed as follows: group A (3.0-3.5 mm, n = 15), group B (3.5 3.75 mm, n = 64), and group C (3.75-4.5 mm, n = 24). RESULTS: At post intervention, average stent area increased by the balloon size (A: 7.2 +/- 1.4 mm2, B: 8.9 +/- 1.5 mm2, and C: 12.1 +/- 2.0 mm2, respectively p < 0.001). At 6 month follow up, the average lumen area increased by balloon size (A: 4.1 +/- 1.7 mm2, B: 5.7 +/- 1.2 mm2, and C: 8.1 +/- 2.0 mm2, respectively p < 0.001). Accordingly, group B and C revealed lower restenosis, compared to group A (A: 46.7%, B: 10.9%, C: 8.3%, A vs B: p = 0.001; C vs A: p = 0.015). Moreover, target lesion revascularization was less in group B and C than group A (A 26.6%, B 3.1%, C 0%, A vs B: p = 0.011; C vs A: p = 0.017). CONCLUSIONS: For non-small vessels, IVUS-guided BMS implantation showed less restenosis and target lesion revascularization compared to small vessels, mainly due to larger initial gain. These study results suggest that IVUS-guided optimal BMS implantation for selected lesions might result in favorable long-term outcomes similar to those seen using drug-eluting stents. For a decade, coronary stenting has become a standard therapy for coronary artery disease due to favorable long-term outcomes and simple treatment procedure. Furthermore, for the last two years, drug-eluting stents (DES), releasing antiproliferative agents from bare metal backbone, revealed the restenosis rates less than half of those seen using conventional bare metal stents (BMS). While target lesions especially suitable for DES continue to be identified, earlier BMS studies showed that larger target vessel or greater stent area contributed to less restenosis. Thus, optimal IVUS-guided BMS implantation for selected lesions might achieve DES-like long-term outcomes. This study was designed to examine the long-term outcomes of BMS with intravascular ultrasound (IVUS)-guided optimization, using coronary angiography and IVUS data. PMID- 15819303 TI - Assessment of coronary artery disease with nicorandil stress magnetic resonance imaging. AB - BACKGROUND: Although dipyridamole and adenosine have been used as vasodilator agents, we believe they are inadequate for vasodilator perfusion magnetic resonance imaging, due to adverse effects (flushing, warmth, headaches, and arrhythmia). Nicorandil, a potassium channel opener, has been reported to increase coronary blood flow and it was associated with fewer adverse effects than adenosine or dipiridamole. We set out to investigate whether the coronary artery stenosis could be assessed by nicorandil stress perfusion magnetic resonance imaging. METHODS: First-pass contrast-enhanced magnetic resonance images of the left ventricle acquired from 50 patients at rest and during intravenous administration of nicorandil using multi-slice turbo field echo with multi shot echo-planar-imaging. Coronary angiography was performed within 1 week. RESULTS: There was no adverse effects during nicorandil stress in any patients. The overall sensitivity and specificity of magnetic resonance imaging in identifying patients with significant stenosis of at least one coronary artery were 93.9% (31 of 33 patients) and 94.1% (16 of 17 patients), respectively. The sensitivity of magnetic resonance imaging for detecting significant stenosis in the left anterior descending artery was 87.5%; the sensitivity in the left circumflex artery was 80%; the sensitivity in the right coronary artery was 92.3%. Similar sensitivities were observed for all 3 vascular regions, indicating that all myocardial segments were visualized with similar image quality. CONCLUSIONS: The present study shows that nicorandil stress perfusion magnetic resonance imaging is a safe, feasible technique for assessing coronary artery stenosis severity in a totally-noninvasive manner. PMID- 15819304 TI - Nonoprerative management of obstructive jaundice caused by a benign hepatic cyst. AB - A 64-year-old man with a 9.0 cm benign hepatic cyst near the hepatic hilum presented with obstructive jaundice caused by the compression of the hepatic ducts. The cyst was aspirated percutaneously, and minocycline hydrochloride was instilled as a sclerosant through a catheter inserted into the cyst. The cyst gradually became smaller, relieving the obstructive jaundice. There were no complications or side-effects. The instillation of minocycline hydrochloride is a useful alternative to ethanol instillation for treating benign hepatic cysts, even when the cyst is causing obstructive jaundice. PMID- 15819305 TI - A neurocognitive and socioecological model of self-awareness. AB - In the past, researchers have focused mainly on the effects and consequences of self-awareness; however, they have neglected a more basic issue pertaining to the specific mechanisms that initiate and sustain self-perception. The author presents a model of self-awareness that proposes the existence of 3 sources of self-information. First, the social milieu includes early face-to-face interactions, self-relevant feedback, a social comparison mechanism that leads to perspective taking, and audiences. Second, contacts with objects and structures in the physical environment foster self-world differentiation in infants; this environment also contains self-focusing and reflecting stimuli, such as mirrors and video cameras. Third, the self can develop bodily awareness through proprioception and can reflect on itself using imagery and inner speech. Furthermore, self-awareness is mainly mediated by the prefrontal lobes. The author establishes various links among these different neurological, social, ecological, and cognitive elements of the model. PMID- 15819306 TI - Reducing the drop-out rates of at-risk high school students: the Effective Learning Program (ELP). AB - The purpose of the present study was to evaluate the effectiveness of the Effective Learning Program (ELP) with students who are at high risk for dropping out of high school. The intervention sought to change external control expectancies to more internal ones, improve students' skills in building relationships with peers and adults, and increase graduation rates. Thirty-eight students considered at risk for dropping out of high school received the intervention. In addition, ELP-eligible students who did not receive the ELP intervention (n = 36) and regular education students (n = 50) from the same high school were compared with the students who received the ELP. A significantly greater percentage of ELP students graduated, became more internally controlled, achieved more as shown by standardized tests, and developed greater social skills and better relationships than their ELP-eligible but nonparticipating peers. The authors suggest that researchers identify which aspects of the ELP are responsible for the improvement in retention, achievement, and personal attributes. PMID- 15819307 TI - Dimensions of premorbid functioning in schizophrenia: a review of neuromotor, cognitive, social, and behavioral domains. AB - After a brief discussion of methods for studying the premorbid period in schizophrenia, the authors review relevant experimental literature to assess the validity of current theories. A critical review is provided that includes representative empirical investigations on premorbid functioning in schizophrenia in the domains of neuromotor, intellectual, academic, cognitive, social, emotional, and behavioral functioning. Limitations of past research, such as lack of specificity to schizophrenia populations, high incidences of false positives, questionable definitions of the premorbid period, inconsistent measures of premorbid functioning, and problems with the current method of DSM classification are discussed. Alternative approaches to studying early functioning in schizophrenia are highlighted. PMID- 15819308 TI - Nurses must use their voice for the good of nursing. PMID- 15819309 TI - We may lose dementia drugs that work. PMID- 15819310 TI - Practice-based commissioning: what is it? PMID- 15819311 TI - A strong and flexible nursing community. PMID- 15819312 TI - Modernising prison mental health care. AB - Some 90% of prisoners in England and Wales have a mental health poblem. This article outlines several initiatives, including health screening on admission and suicide prevention, to improve the care of this population. PMID- 15819313 TI - A centre for advice and treatment. AB - The first wave of NHS walk-in centres opened in 2000. Since then, a total of 57 centres have opened in England with 25 more planned in the next few months. These centres do not aim to replace existing primary or secondary care services but to complement them and provide the public with an alternative means of accessing health care for minor illnesses, minor injuries and emergency contraception. Two models of walk-in centre exist: those based in primary care and those based in secondary care. PMID- 15819314 TI - Diabetes and nutrition: the latest thinking on dietary management. AB - Dietary management is an important part of the care of people with both Type 1 and Type 2 diabetes. This article examines the latest thinking on diet, including use of the plate model, based on recommended percentages of foods from the five food groups, and dispels some common myths about food and diabetes. PMID- 15819315 TI - Improving the management and quality of life of patients with allergic rhinitis. AB - Allergic rhinitis affects up to a fifth of the population in the West, yet there is little specialist care to support and manage patients, and medical and nursing curicula fail to address the issue of allergies. This paper looks at the main causes of allergic rhinitis and its symptoms, and describes the available treatments to manage the conditon. PMID- 15819316 TI - The use of honey-derived dressings to promote effective wound management. AB - Clinical observations suggest that honey holds significant promise as an effective treatment for a number of medical conditions and particularly in the management of non-healing wounds. This article explores the healing properties of honey and illustrates the effectiveness of using honey in the management of chronic leg ulcers. PMID- 15819317 TI - Reducing the risks related to the handling and disposal of health-care waste. AB - Guidelines on the management of hospital waste aim not only to minimise the risks posed to staff at all levels but also to contain potential environmental hazards, such as infection and sharps injuries. It is therefore important for all personnel to be vigilant and to ensure that they dispose of clinical waste safety and appropriately. PMID- 15819318 TI - Diet will help reduce cholesterol levels. AB - A cardio-protective diet, rich in omega-3 oils and fresh fruit and vegetables, will help lower cholesterol levels. In this article, a dietitian provides answers to frequently asked questions and advice on good dietary practice. PMID- 15819319 TI - How the use of electric profiling beds can reduce the prevalence of pressure ulcers. AB - Investment in electric profiling beds and a change in patient-assessment procedures have had a significant impact on the prevalence of hospital-acquired pressure ulcers in one trust, while at the same time increasing patient independence and reducing pain, length of hospital stay and the risk of potentially costly litigation. PMID- 15819320 TI - Pharmacological treatments for obesity. AB - Carol McLoughlin looks at the advantages and drawbacks of using drugs to treat obesity and describes new areas of research that may offer new solutions to tackling this rapidly growing health and medical problem. PMID- 15819321 TI - Reshaping delivery of care. PMID- 15819322 TI - Interpreting policy in the community. Interview by Carolyn Scott. PMID- 15819323 TI - Not knowing is the worst thing. PMID- 15819324 TI - Testing research. PMID- 15819325 TI - Tokyo story. A British nurse is setting up an infection control service in Japan. AB - Tokyo is an amazing capital city, just as you would expect: tall buildings, bright lights, overcrowded and fast paced. Japan was a bit of a culture shock at first, but things have settled down since I learned a few key Japanese phrases for such questions as 'Can I have... ?' and 'Where is... ?' There are many places to visit in this city of almost 15 million people. Getting around is straightforward, as there is an efficient underground system with the station names in English as well as Japanese. The trains are on time and fully air conditioned, but during rush hours it can get very uncomfortable with so many people squashed inside the carriages. The food ranges from inexpensive pavement cafes serving batter-fried octopus to exclusive sushi restaurants. Every type of food is available in the many restaurants, but I enjoy trying traditional Japanese dishes. The food is always fresh, well presented and tasty. Some of the more unusual dishes include raw horse, raw eggs, sea urchins and live fish. I have been taken to a karaoke bar by my work colleagues and it was great, but very different from what I had expected. You only sing to the people you are with and there are a number of rooms in the bar, depending on how many of you there are, which reduces the public humiliation factor. PMID- 15819326 TI - In the public interest. PMID- 15819327 TI - One jab benefits all. Assessing the impact of extending the vaccination programme. PMID- 15819329 TI - Face the fats. PMID- 15819330 TI - Lost generation. Student role models are disappearing fast. PMID- 15819328 TI - On the trail. Three parlimentary nurse hopefuls outline their personal manifestos. Interviews by Alison Moore. PMID- 15819331 TI - Self-assessment and reflection in nurse education. AB - It is widely argued that nurse learners should learn to become reflective practitioners and that self-assessment plays a part in this (Ekebergh et al 2004, Thorpe 2004). Problems arise, however, when consideration is given to just how this right be achieved. This article examines the challenges of assessing reflection where self-assessment has played a part. It highlights the challenges that can arise when reflection and self-assessment are used for different educational purposes. It identifies why this work is challenging and suggests some parameters that may assist colleagues in making learning more successful. PMID- 15819332 TI - Common food allergies. AB - The incidence of allergic disease, including food allergy, appears to be increasing in the UK (Gupta et al 2003). Although any food has the potential to cause an allergic reaction, certain foods are more common causes of allergy than others. If diagnosed, food allergy is manageable. Correct diagnosis is important to ensure optimal management and a nutritionally balanced diet. PMID- 15819333 TI - Diet and menopausal health. AB - This article outlines the evidence for exercise and dietary influences on many of the major disease risks for women around the menopause and after, including cardiovascular diseases, cancer and osteoporosis. PMID- 15819334 TI - Leading the charge. The role of NVQ facilitator. PMID- 15819335 TI - Building a collaborative research agenda: drug abuse and HIV/AIDS in the Caribbean 2002-2004. PMID- 15819336 TI - When pigs fly. Confronting the new era of disease transmission. PMID- 15819337 TI - Changing the perception of caries. PMID- 15819338 TI - Heart rate variability. PMID- 15819339 TI - Parecoxib versus ibuprofen. PMID- 15819340 TI - Dental entrepreneurship. PMID- 15819341 TI - Fluoride varnish application. PMID- 15819342 TI - Voices from the front: dentists share their stories of war. PMID- 15819343 TI - Dr. John Caulfield's report from the front. PMID- 15819344 TI - A three-year clinical evaluation of two-bottle versus one-bottle dentin adhesives. AB - BACKGROUND: The authors conducted an in vivo investigation to compare the clinical performance of two commercial one-bottle adhesives and a two-bottle adhesive for restoration of noncarious cervical lesions (NCCLs). METHOD: The patient pool consisted of 57 patients and 171 teeth (three teeth per patient), with one NCCL per tooth. Each patient received three resin-based composite restorations, each with a different adhesive: one tooth with a two-bottle, water based adhesive as the control; another tooth with a one-bottle, ethanol-based adhesive; and a third tooth with a one-bottle, solvent-free adhesive. The authors assessed restorations in terms of retention, marginal integrity, margin discoloration and air sensitivity at baseline, six months, one year, two years and three years after initial placement. RESULTS: The retention rates at 36 months were 88 percent for the first adhesive, 81 percent for the second adhesive and 90 percent for the third adhesive. No statistically significant differences in retention rates could be shown, with 86 percent of restorations retained overall. Measures of marginal integrity, marginal discoloration and sensitivity also had no statistically significant differences between the three adhesives (P > .05). CONCLUSIONS: All three adhesives performed with acceptable outcomes after a 36-month period, with small differences between the one- and two-bottle systems and between the various solvents. Retention rate was moderately high and air sensitivity was markedly reduced; however, superficial marginal discoloration and marginal degradation was notable. Certain lesion, tooth and patient characteristics may predispose restorations to retention failure. CLINICAL IMPLICATIONS: The type of solvent may not be a major factor in retention of Class V restorations in NCCLs. Both single-bottle adhesives and conventional two-bottle adhesives performed acceptably. PMID- 15819345 TI - Assessing the accuracy of caries diagnosis via radiograph. Film versus print. AB - BACKGROUND: The authors conducted this study to determine if proximal caries diagnoses made using bitewing radiographic images printed on photographic paper were comparable with diagnoses made using traditional radiographic film images. METHODS: The authors digitized 15 posterior bitewing radiographs that contained 74 carious and 127 sound unrestored proximal surfaces and printed them on photographic paper. Fourteen dentists evaluated the radiographs and two printed image formats (4 x 3 centimeters and 8 x 6 cm) for evidence of caries. The diagnostic accuracy and interobserver agreement for caries diagnoses obtained in the two printed image formats were compared with those for radiographic film images. RESULTS: Overall, the diagnostic accuracy of printed images did not differ significantly from radiographic film images for dentinal caries. However, for caries limited to the enamel surface, a decrease in sensitivity was noted in six of the 14 observers for the smaller print images, while no significant differences in the diagnoses of enamel caries were observed among any of the observers in the enlarged print format. CONCLUSION: This study provides evidence that printed images can be used to diagnose dental caries reliably. CLINICAL IMPLICATIONS: The results of this study indicate that the diagnostic information obtained by viewing printed images is equivalent to that obtained by viewing standard radiographs. Size of the printed image also may be important in caries diagnosis and care must be taken to print bitewing radiographic images at a size that optimizes interpretation. Other factors that must be considered are the type of printer, printer resolution, paper quality and type of ink used. With careful consideration of printing parameters, clinicians can be assured of diagnostic quality in printed images. PMID- 15819346 TI - The clinical significance and management of apical accessory canals in maxillary central incisors. AB - BACKGROUND: The maxillary central incisor is considered to be the least difficult subject for a clinical endodontic experience. However, the internal anatomy of maxillary central incisors can present a number of variations, including multiple accessory canals. CASE DESCRIPTION: This article highlights the clinical significance and management of accessory canals located in the apical one-third of maxillary central incisors. The authors present two case reports in which failure to detect the accessory canals led to root canal failure and subsequent surgical intervention. Another two case reports present the serendipitous discovery and nonsurgical management of accessory canals during the initial treatment of maxillary incisors. CONCLUSIONS AND CLINICAL IMPLICATIONS: It is important for the clinician to be able to detect the signs suggesting the presence of accessory canals in maxillary central incisors. Failure to do so may lead to a less-than-optimal endodontic treatment outcome. PMID- 15819347 TI - Root resorption and ankylosis associated with guided tissue regeneration. AB - BACKGROUND: Root resorption and ankylosis have been reported rarely as sequelae to guided tissue regeneration (GTR). The authors describe a clinical case of root resorption following GTR that involved the use of a bioabsorbable membrane. CASE DESCRIPTION: Two years after GTR was performed on a Class II furcation defect, the clinical examination revealed root resorption reaching the pulp chamber. The furcation defect was filled with epithelium and connective tissue, which contained inflammatory infiltrate and fragments of the membrane. The authors also observed areas of the tooth that exhibited points of ankylosis and root resorption. CLINICAL IMPLICATIONS: Clinical trials have reported favorable clinical and histologic results with GTR. However, this case report, along with other case reports and studies in animals, suggests a high risk of root resorption and ankylosis after GTR, which could limit the indications for this technique. PMID- 15819348 TI - The state of fixed prosthodontic impressions: room for improvement. AB - Impressions for crowns and fixed prostheses could be better. I have offered several suggestions to improve their quality. None of the suggestions is difficult or time-consuming. The result of making better impressions will be greater longevity for the resultant restorations and happier patients, dentists and laboratory technicians. PMID- 15819349 TI - The patient with a heart murmur: evaluation, assessment and dental considerations. AB - BACKGROUND: Heart murmurs, a common finding in dental patients, are of major concern to dental professionals because certain dental procedures occasionally can induce severe cardiovascular complications. Murmurs may indicate existing heart disease that is a risk factor for infective endocarditis following a dental procedure, as well as more severe heart conditions such as congenital heart disease, atrial fibrillation or congestive heart failure. TYPES OF STUDIES REVIEWED: This review article is based on data published in peer-reviewed journals, including practice guidelines published by major dental and medical professional organizations. RESULTS: Echocardiography is the primary means of evaluating heart murmurs, and all dental professionals should become familiar with major aspects of an echocardiogram. Understanding the medical evaluation and assessment of a heart murmur fosters better communication with other health care professionals and results in improved patient care. CLINICAL IMPLICATIONS: Beyond the need to administer antibiotic prophylaxis, the dentist also needs to address the underlying causes of a patient's heart murmur. By providing dental care to such patients, oral health care providers become part of the patient's overall health care team. PMID- 15819350 TI - Dental practice: prices, production and profits. AB - BACKGROUND: This article explores the relationship of dental fee increases, physicians' fee increases and overall price increases. The author uses the applicable consumer price indexes (CPIs) for the past 30 years to determine whether any similarities in behaviors exist. The relationship between office operating costs, production and net income also are explored. CONCLUSIONS: Although there are quantitative differences, the CPIs move in parallel fashion. An abrupt change in the overall trend of all CPIs to a decrease in the degree of increases occurred in the 1980s and has continued. Increased production by dental offices is the key strategy being used to maintain office net income. PRACTICE IMPLICATIONS: Dentists would be wise to track the CPIs for trend information that could be useful to them as they manage their practices and devise strategies to maintain net income. PMID- 15819351 TI - Measuring patient satisfaction. AB - Many businesses use customer satisfaction surveys successfully. You may notice that you find one in almost every restaurant or hotel room. I do not think it is a coincidence that the hotel industry provides some of the finest customer service available. When it comes to providing excellent customer service, dental practices can learn from businesses that regularly assess customer satisfaction. PMID- 15819352 TI - Parental perceptions of their preschool-aged children's oral health. AB - BACKGROUND: Parents have an important role in making decisions about their children's oral health. The purpose of the authors' study was to determine parental perceptions of their children's oral health status and factors correlated with these perceptions of health. METHODS: The authors analyzed data for 3,424 children (2-5 years of age) from the Third National Health and Nutrition Examination Survey. They based the dependent variable on a question asked of primary caregivers: "How would you describe the condition of [child's name]'s natural teeth?" Explanatory variables included demographic variables, dental visits, perception of child's general health, need for dental care and presence of tooth caries. RESULTS: Eighty-nine percent of parents rated their child's oral health as excellent, very good or good, and 11 percent rated it as fair or poor (mean = 2.7 on a five-point scale, with 1 being excellent and 5 being poor). Tooth caries, perceived need for dental cleaning and treatment, lower income and poorer general health perceptions were associated with poorer parental ratings. CONCLUSIONS: Actual disease and perceived need are associated significantly with parents' perceptions of their children's oral health. PRACTICE IMPLICATIONS: Understanding parents' perceptions of their children's oral health and factors that motivate these perceptions can help dentistry overcome barriers that parents encounter in accessing dental care for their children. PMID- 15819353 TI - Assessing oral cancer knowledge among dental students in South Carolina. AB - BACKGROUND: Because South Carolina has the fourth highest mortality rate for oral cancer among the 50 states, dental students in the state must be knowledgeable about prevention and early detection of the disease. METHODS: In 2002, the authors surveyed 163 students using a written questionnaire (response rate, 79.1 percent). The questionnaire included questions about oral cancer risk and nonrisk factors as well as oral cancer diagnostic signs, symptoms and examination procedures. The authors performed univariate and bivariate analyses (alpha < or = .025). RESULTS: At least 93 percent of the students replied that tobacco, alcohol and previous oral cancer lesions were risk factors. One hundred six students (65 percent) knew that the most likely site for oral cancer is the ventrolateral border of the tongue. Students differed in their overall knowledge of risk factors (P = .002), nonrisk factors (P < .001) and diagnostic procedures (P < .001). CONCLUSION: Although students' level of knowledge increased with academic year, educators and policy-makers need to place greater emphasis on oral cancer education and training in dental schools. PRACTICE IMPLICATIONS: Morbidity and mortality are likely to be reduced if dentists know how to prevent and detect oral cancer. PMID- 15819354 TI - The clinical performance of professionally dispensed bleaching gel with added amorphous calcium phosphate. AB - BACKGROUND: The authors undertook a study to measure how the addition of amorphous calcium phosphate (ACP) to a professionally dispensed 16 percent carbamide peroxide equivalent bleaching gel affects tooth color and dentinal hypersensitivity. METHODS: The authors assigned two groups to use either the test gel containing ACP or a control gel. Both groups used their respective products for three hours daily for 14 days. At checkpoints during the treatment period, the authors studied tooth color, gingival health and three measures of hypersensitivity. They performed double-blinded clinical measurements on days three, seven, 14 and on the fifth day post-treatment. RESULTS: The test group demonstrated significantly lower (P < .05) mean thermal sensitivity scores compared with baseline (day 14: 0.21 versus 0.31; fifth posttreatment day: 0.06 versus 0.18). Tactile sensitivity also was substantially lower (P < .05) for test subjects (day 14: 0.26 versus 0.48; fifth posttreatment day: 0.06 versus 0.19). Furthermore, at the conclusion of the study, twice as many subjects were free of thermal sensitivity (test group, 80 percent, compared with control group, 40 percent; P < .001) and there was a similar significant (P < .001) percentage difference for tactile sensitivity. Both groups demonstrated equivalent and significant tooth color enhancement as compared with baseline (control: -7.73 shade change versus test: -8.12; P < .05). CONCLUSIONS: This study demonstrates that ACP could be added to a 16 percent carbamide peroxide equivalent bleaching gel and result in a significant reduction of clinical measures of dentinal hypersensitivity, both during and after treatment. CLINICAL IMPLICATIONS: The results of this study offer evidence in support of clinical decisions to treat patients with bleaching gel containing ACP when uncompromised tooth whitening efficacy is desired, yet dentinal hypersensitivity may be a concern. PMID- 15819355 TI - Antitrust revisited: the pitfalls of the messenger model. PMID- 15819356 TI - Extreme makeovers. PMID- 15819357 TI - For the dental patient... Canker sores and cold sores. PMID- 15819358 TI - Ten years of family change. PMID- 15819359 TI - School lunches: England starts to improve standards. PMID- 15819360 TI - Atopic eczema: nurse-led care--1. Making the most of the consultation. AB - Nurses are now increasingly involved in diagnosing, prescribing and initiating treatments for clients with eczema. In the first of a two-part article, a nurse consultant in dermatology focuses on the clinical criteria for deciding whether the client has eczema and advises on how both nurse and client can make optimum use of the consultation. Ways of assessing clients' eczema and its impact on their daily life are discussed. Part 2 of the article, to be published in the next issue of Journal of Family Health Care, will examine treatments for eczema. PMID- 15819361 TI - Healthier lifestyles series: 1. Exercise for children. AB - Statistics indicate that 39% of boys and 58% of girls aged 7-18 do not achieve the recommended levels of exercise, eg spending at least one hour each day in a physical activity of at least moderate intensity. This paper summarises the physical and psychological health benefits of exercise for children and young people and emphasises that the promotion of exercise and healthy food choices should go hand in hand. Practical tips for young people on exercise and nutrition are included. PMID- 15819362 TI - Sexually transmitted infections. Genital chlamydia: practical management in primary care. AB - As the number of infected people increases, more nurses outside the specialist sexual health services are being called upon to provide advice and clinical care for genital chlamydial infection. This article provides non-specialist nurses with practical information on the diagnosis, treatment, management and follow-up of clients with genital chlamydial infections. PMID- 15819363 TI - "A worrying year for community child health: roll on, 2005". PMID- 15819364 TI - School nursing: your views are wanted. PMID- 15819365 TI - Differences in child rearing. Cultural contrasts: bringing up children the Honduran way. AB - The author, a health visitor now living in Honduras, Central America, describes child-rearing practices in Honduras and contrasts them with those in the UK. PMID- 15819366 TI - [Image of the month. An endoscopic image of tubal pregnancy]. PMID- 15819367 TI - [Image of the month. Aortic quadricuspid, a rare cause of aortic insufficiency]. PMID- 15819368 TI - [How I treat...varicosities by sclerotherapy]. AB - Sclerotherapy is a technique for the treatment of varicosities, vascular ectasia, and sometimes, leg varicose veins. This method is aesthetic and in some cases therapeutics and uses sclerosing agents, electrocoagulation and exceptionally LASER. These relatively simple methods are safe and efficient if they are applied with a rigorous procedure by well trained physicians. PMID- 15819369 TI - [Clinical case of the month. Peritonitis after spontaneous rupture of pyonephrosis. A case report]. AB - Peritonitis after spontaneous rupture of pyonephrosis is a rare complication, usually diagnosed intraoperatively. We report the case of a 35 year old woman who was admited for an acute abdomen. Ultrasound showed features of acute peritonitis, with left pyonephrosis, confirmed by a CT-SCAN. Treatment consisted of laparotomy with nephrectomy and abdominal washing and draining. The outcome was favourable. PMID- 15819370 TI - [The "top" of cardiology literature in 2004: an annual selection of the American Heart Association]. AB - The American Heart Association has once again released its top ten research advances in heart disease and stroke for the past year. As always, the selection is assorted. It contains very pointed, at times preliminary, research, as well as major clinical trials; it also reports findings that seem of particular interest for an American audience or come as a support for a special programme proposed by the association. Reading these ten abstracts is, however, invariably rich of enlightenment, and stimulation for everyone. PMID- 15819371 TI - [PPAR-gamma receptors, new therapeutic target in metabolic and cardiovascular diseases]. AB - PPARs ("Peroxisome Proliferator-Activated Receptors") belong to a superfamily of nuclear receptors with several isoforms, among which PPAR-alpha mainly located in the liver and PPAR-gamma mainly located in the adipose tissue. These receptors are considered as major pharmacological targets since the discovery of their activation by specific agonists, which lead to various favourable metabolic effects. Improvement of lipid profile by fibrates is explained by the activation of liver PPAR-alpha receptors. However, PPAR-gamma receptors have focused most fundamental and clinical research in recent years after the demonstration of their activation by thiazolidinediones (pioglitazone, rosiglitazone), a new class of antidiabetic agents. Beyond their effects on insulin sensitivity, glitazones exert pleiotropic effects that may result in cardiovascular protection in high risk patients. It has been recently demonstrated that certain angiotensin AT1 receptor blockers (sartans) can also exert a partial agonist activity on PPAR gamma. Among the molecules of this class, telmisartan appears to exert this effect at the lower concentrations. Thus, PPAR-y, as common pharmacological target, may, at least partially, explain some of the effects observed with both thiazolidinediones and inhibitors of the renin-angiotensin system, in particular the improvement in insulin sensitivity (in particular via an increase in adiponectin levels), the protection against type 2 diabetes, the reduction in arterial blood pressure and the prevention of cardiovascular complications. There is currently a major interest from the pharmaceutical industry in the development of new molecules able to activate both PPAR-alpha and PPAR-gamma. PMID- 15819372 TI - [Primary orthostatic tremor]. AB - Primary orthostatic tremor is a particular tremor exclusively present when a subject is standing. Patients experience a severe disabling sense of unsteadiness. Walking, sitting and lying down are unaffected Neurological examination and cerebral imagery are normal most of the time. Electromyography in standing position confirms the diagnosis in showing a regular rapid tremor with a frequency of 12 to 18 Hz. Its physiopathology is partially unknown. A few symptomatic therapies can be proposed. PMID- 15819373 TI - [Clinical value of cardiac resynchronization therapy in patients with congestive heart failure]. AB - Besides major progress in the pharmacologic treatment of severe chronic heart failure, cardiac resynchronization therapy (CRT) has developped over the last ten years. We report the follow-up of the 36 first patients with a CRT device implanted from July 2000 to November 2002 at the CHR de la Citadelle Hospital in Liege. After a mean follow up of 6 months, no death was observed. The functional benefit of resynchronization is validated by a significant improvement in the NYHA class, an increase in the walking distance measured by the six minute walk test from 268 +/- 103 to 342 +/- 106 meters (p < 0.004) and by a not significant rise in the VO2 max (from 11.1 +/- 2.8 to 14 +/- 10 ml/kg/min; P=0.1). The quality of life, assessed by the Minnesota-Living-In-CHF score, improves from 49 +/- 20 to 35 +/- 22 after the six month follow-up (P=0.02) The echocardiogram also shows a better left ventricular ejection fraction at six months, from 24 +/- 7% to 31 +/- 7% (P<0.05). Based on a better NYHA functional class, responders (n=24; 71%) and non responders (n=10; 29%) were compared; a correlation between the functional class change and the improvement of the ejection fraction was documented, but not with the reduction in QRS width. Our registry, with the potential pitfalls of a monocentric prospective study, confirms the feasability, safety and efficacy of CRT in severe chrbnic heart failure uncompletely corrected pharmocalogically. It remains however approximately 30% of non responding patients, in whom the current clinical studies should help identify the right criteria to predict and discriminate responders. PMID- 15819374 TI - [Compartment syndrome and sport traumatology]. AB - Compartment syndrome is defined as an increased intracompartmental pressure within inelastic fascia which surround muscular compartments. That pathology can be either acute or chronic. The acute situation is generally a medical emergency. Unless a fasciotomy is rapidly performed, increasing pressure may completely cut off blood flow and cause necrosis of the affected limb. The chronic compartment syndrome, more frequent, is characterized by exercise-induced pain and swelling that is relieved by rest. It results from an excessive intra-muscular pressure increase. The anterior compartment of the leg is most commonly involved. To accurately diagnose the compartment syndrome, physicians perform measurements of compartment pressure. By means of miniature digital instruments, exercise and rest intramuscular pressure are recorded. If pressures reach the critical threshold (30 mm Hg) and remain high five minutes after exercise, compartment syndrome is present. Treatment is nearly exclusively surgical. It consists in an fasciotomy allowing a complete recovery and a return to sport activities. PMID- 15819375 TI - [How I explore...a disorder with an apparently healthy microscopic aspect of the skin]. AB - Some dermatologic disorders named "Invisible dermatoses" are defined by obvious clinical signs while histologic alterations remain discrete. This situation may lead to clinico-pathological discrepancies impeding the establishment of a diagnosis. In other cases, the skin looks clinically normal but the histological examination allows to disclose some systemic diseases. Systematic analysis of the skin biopsy complemented by histochemical and immunohistochemical techniques may help reach the diagnosis. PMID- 15819376 TI - [Bariatric surgery: 10-year results of the Swedish Obese Subjects Study]. AB - The 10-year results of the prospective, controlled Swedish Obese Subjects Study were recently reported in the New England Journal of Medicine by L. Sjostrom and colleagues. This trial compared obese subjects who underwent gastric surgery and contemporaneously matched, conventionally treated obese control subjects. The follow-up rate for laboratory examinations was 74.5 percent at 10 years. At that time, data of 627 patients of the control group (mean age of 48 years, body mass index of 41 kg/m2) were compared to those of 641 patients who were submitted to surgery (banding n = 156, vertical banded gastroplasty n = 451 and gastric bypass n =34). At 10 years, the body weight had increased by 1.6 percent in the control group and decreased by 16.1 percent in the surgery group (p < 0.001), and similar changes were observed for waist circumference (+2.8 percent versus -10.1 percent, respectively, p < 0.001). Energy intake was lower and the proportion of physically active subjects higher in the surgery group than in the control group throughout the observation period. Ten-year rates of recovery from diabetes, hypertriglyceridaemia, low levels of high-density lipoprotein cholesterol, hypertension, and hyperuricaemia were more favourable in the surgery than in the control group. The surgery group had lower 10-year incidence rates of diabetes, hypertriglyceridaemia, and hyperuricaemia (but not of hypertension) than the control group. In conclusion, as compared with conventional therapy, bariatric surgery appears to be a valuable option for the treatment of severe obesity, resulting in long-term weight loss, improved lifestyle, and, except for hypercholesterolaemia that was not significantly affected, amelioration in cardiovascular risk factors that were elevated at baseline. Obtaining long-term data concerning the effect of weight loss on overall mortality and on the incidence rates of myocardial infarction, stroke, and cancer remains a key objective of this landmark study. PMID- 15819377 TI - Cardiovascular prevention in type 2 diabetic patients: review of efficacious treatments. AB - BACKGROUND: Type 2 diabetes (t2DM) is a chronic and complex metabolic condition requiring continuing medical care in order to reduce the risk of long-term complications. Macrovascular complications cause about 65% of deaths in subjects with t2DM and are responsible for severe co-morbidity. Many studies have addressed cardiovascular (CV) risk reduction in t2DM subjects. OBJECTIVES: To summarise the evidence concerning the impact of lifestyle and of medical interventions in t2DM patients on CV risk (myocardial infarction, stroke, CV death, or a combination of these). METHODS: We successively reviewed the recent guidelines addressing CV prevention in t2DM and searched the Cochrane Controlled Trials Register (CCTR), Medline & Embase to find systematic reviews and original articles on CV events in t2DM patients. We selected original studies which included solely t2DM patients or a large t2DM subgroup (n>100), tested lifestyle habits or drug treatments, and analysed CV endpoints. Their design had to be a randomised controlled trial for drug interventions, and a prospective cohort for lifestyle habits. MAIN STUDIES: We found 4 major guidelines focusing on CV prevention in t2DM patients, all released in 2003, and 10 reviews and meta analyses: one dealing with dietary intervention, three with blood pressure lowering, one with ACE-inhibitors, one (with update) with platelet-inhibitors , three with cholesterol-inhibitors and one that dealt with both cholesterol inhibition and blood pressure lowering. We included cohort studies on cigarette smoking (1), physical exercise (3) and weight control (1), as well as randomised trials on treatment of glycaemia (1), lipidaemia (13), blood pressure (12) and platelet aggregation (4). We also included one open randomised trial dealing with a multifactorial treatment. CONCLUSIONS: Global CV risk management in t2DM should aim at changes in lifestyle habits and daily use of multiple drugs. Treatment should be long-term and target-driven with intensified interventions aimed at all validated targets. Lifestyle approach is of primary importance. Five targets are supported by strong clinical evidence (Table 4): reduction of blood pressure and of LDL-Cholesterol to normal values, and the use of three types of drugs which inhibit, respectively, platelet aggregation, angiotensin pathway and cholesterol synthesis. PMID- 15819378 TI - Patterns of physical activity defined by continuous heart rate monitoring among children from Liege. AB - Health benefits of a physically active lifestyle are well documented. We therefore investigated the physical activity patterns of 200 children from Liege. They were monitored continuously using a 24-hour Holter monitoring system during normal weekdays and the percentage of heart rate reserve (%HRR) was used to measure the amounts of physical activity at different intensities. Preschool children attained 184.3+/-54.2, 40.7+/-16.1, 15.8+/-6.9 and 6.0+/-7.2 minutes/day (mean+/-SD) between 20% to 40%, 40% to 50%, 50% to 60%, and greater than 60% of HRR, respectively. At the same %HRR intensities, schoolchildren attained 165.6+/ 74.6, 32.1+/-12.1, 15.8+/-6.7 and 7.0+/-5.9 minutes/day, and teenagers attained 159.2+/-68.3, 32.1+/-23.5, 13.1+/-6.0 and 6.1+/-6.3 minutes/day. Age was a significant predictor of the intercept and slope of the time spent in physical activity and %HRR relationship. In Liege the average youth accumulates +/-30 to 40 minutes/day of moderate-intensity physical activity and +/-20 minutes/day of high-intensity physical activity. Those children meet the classical revised guidelines for physical activity but do not compare favourably with children from elsewhere. On the other hand, they get more than 2 1/2 to 3 hours/day of low intensity physical activity. Our findings suggest that children from Liege are not engaged in sedentary behaviour but do not experience the ideal amount and type of physical activity classically believed to benefit the cardiopulmonary system. Public health strategies should be adapted to our findings. PMID- 15819379 TI - Cost consequence analysis of fondaparinux versus enoxaparin in the prevention of venous thromboembolism after major orthopaedic surgery in Belgium. AB - Fondaparinux, a selective inhibitor of activated factor X, has been shown to reduce further the risk of venous thromboembolism (VTE) in major orthopaedic surgery compared to the low molecular weight heparin enoxaparin, when both were applied for 7 days after surgery. To compare the expected costs and clinical outcomes of fondaparinux with enoxaparin applied for 7 days after surgery, we conducted a cost-consequence analysis in patients undergoing major orthopaedic surgery, i.e. total hip replacement, total knee replacement and hip fracture repair. Our decision model included endpoints relevant in routine clinical practice and the natural history of VTE over a long term period of 5 years. Costs for prevention, diagnosis and treatment of VTE and its complications were estimated from the Belgian health care payer perspective. Analyses were conducted for different time horizons and for the three indications, separately, and then combined. Overall, our results indicated that the initial investment in fondaparinux (cost per day: 10.39 euros versus 3.74 euros for enoxaparin) was soon compensated by savings due to avoided VTE events, with cost neutrality being achieved after 90 days and further savings being incurred over longer time periods mainly due to avoided post-thrombotic syndromes. These findings were most pronounced in patients undergoing hip fracture repair. Sensitivity analyses showed these findings to be robust for the three indications separately, and combined. We conclude that our analysis of health and economic consequences over a long term period, demonstrates the value for money of fondaparinux versus enoxaparin for the prevention of VTE events after total hip replacement, total knee replacement and hip fracture repair. PMID- 15819380 TI - Anaphylaxis to an urethral lubricant: chlorhexidine as the "hidden" allergen. AB - We report on a severe allergic reaction during urethral catheterisation, initially erroneously attributed to natural rubber latex (NRL). However, the negative investigations for latex, the uneventful further applications of NRL, the positive skin test and basophil activation test for chlorhexidine strongly support diagnosis of chlorhexidine anaphylaxis. This case illustrates that it is not widely appreciated that "innocent" products such as urethral lubricants may elicit serious life-threatening reactions. Careful inspection of the medical records is once again re-emphasised to be mandatory in order to avert subsequent anaphylaxis. PMID- 15819381 TI - Severe hypophosphatemia in a patient with anorexia nervosa during enteral refeeding. AB - Hypophosphatemia is a seldom but potentially fatal complication of the nutritional recovery or refeeding syndrome in patients with protein-calorie malnutrition or starvation. We report here the case of a 35-year-old anorexic patient who presented a severe but uncomplicated hypophosphatemia during enteral refeeding, despite phosphorus supplementation. Serum phosphorus monitoring is recommended in severely malnourished anorexic patients, particularly during the first week of refeeding, be it parenteral or enteral. PMID- 15819382 TI - Staphyloccocal scalded skin syndrome in an adult. AB - We report a case of staphyloccocal scalded skin syndrome due to an oxacillin susceptible Staphylococcus aureus in an 81-year-old woman. The patient was admitted to the emergency room with arthritis of the left shoulder, ten days after an intra-articular injection of corticosteroids. The shoulder's puncture showed a purulent liquid and gram positive cocci in cluster suggesting the presence of Staphylococcus sp. on the Gram-stain. The culture confirmed the identification of an oxacillin-susceptible Staphylococcus aureus. Before administration of any dose of oxacillin, blisters appeared on the skin, that quickly ruptured, particulary in areas of friction. The exfoliated areas were extensive and resolution of all the lesions was reached after 3 weeks. The skin biopsy showed superficial epidermolysis confirming the diagnosis. Staphyloccocal scalded skin syndrome is usually described in neonates and young children, often in outbreaks. Few cases have been reported in adults, most often associated with severe underlying diseases. The mortality rate is low in children but can reach almost 60% in adults. The most important diagnosis to exclude is Lyell's syndrome which can be done by the skin biopsy. PMID- 15819383 TI - Streptococcus milleri-sepsis with lung and brain abscesses. PMID- 15819384 TI - Hepatic necrosis resulting in leak of amiodarone and refractory cardiogenic shock. PMID- 15819385 TI - Pharmacokinetics and disposition of DY-9760e, a novel calmodulin antagonist, in rats and monkeys. AB - DY-9760e (3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1 4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate, CAS 162496-41-5) is a novel calmodulin antagonist that is being evaluated for the treatment of ischemia. The objective of this study was to characterize the pharmacokinetics and disposition of DY-9760e in rats and monkeys. After a 6 h continuous infusion at 1 mg/kg/h to male rats, the plasma concentration of unchanged DY-9760, as the anhydrous free base of DY-9760e, declined with a terminal half-life of 3.0 h. In monkeys, the plasma concentration of DY-9760 following a 4 h continuous infusion at 1 mg/kg/h declined with a terminal half-life of 3.8 h. Total clearance was 18.3 ml/min/kg in rats and 16.7 ml/min/kg in monkeys. The pharmacokinetics of DY 9760e was linear within a dose range from 1 mg/kg to 16 mg/kg in monkeys. After intravenous bolus administration of 14C-DY-9760e to rats, the radioactivity was widely distributed throughout the body except for the brain and testis. In the brain, which is the target organ of this compound, the concentrations of unchanged DY-9760 in rats were much lower than the corresponding plasma concentrations. These results indicated that the permeability of DY-9760 into the brain was restricted. In contrast, the brain concentrations of the N-dealkylated metabolite DY-9836 were approximately 2- to 3-fold higher than those observed in the plasma. The administered radioactivity was excreted mostly in the feces (95.2% in rats, 83.6% in monkeys), and the biliary excretion of the radioactivity in bile duct-cannulated rats was 86.2% within 48 h, part of which (11.1%) was re absorbed. The urinary excretion of unchanged DY-9760 was less than 0.5% in both species. The metabolic profile characterized by thin-layer chromatography demonstrated that most of the radioactivity in the urine and bile referred to many polar metabolites. These results indicate that DY-9760e is eliminated mainly through hepatic metabolic clearance in both rats and monkeys. PMID- 15819386 TI - Interaction of arylpiperazines with the dopamine receptor D2 binding site. AB - The docking of several 1-benzyl-4-arylpiperazines to the dopamine receptor (DAR) D2 was examined. The results demonstrated that the interaction of protonated N1 of the piperazine ring with Asp 86 (III.32) and edge-to-face interactions of the aromatic ring of the arylpiperazine part of the ligand with Phe 178 (VI.44), Trp 182 (VI.48) and Tyr 216 (VII.58) of the receptor, represent the major stabilizing forces. Besides, the hydrogen bond acceptor group in position 2 of the phenylpiperazine aromatic ring could build one more hydrogen bond with Trp 182 (VI.48). Bulky substituents in position 4 were not tolerated due to the unfavorable sterical interaction with Phe 178 (VI.44). Substituents in position 2 and 3 were found to be sterically well tolerated. Introduction of electron attractive -NO2 group in position 3 of aryipiperazines decreased, while electron donors (-OMe) and the second aromatic ring (naphthyl) increased the binding affinity comparing to that of the phenylpiperazine 1. This can be explained in terms of favoured edge-to-face interactions in ligands with a high negative electrostatic surface potential (ESP) in the centre of aromatic residue of arylpiperazines. Thus, besides the salt bridges and hydrogen bonds, edge-to-face interactions significantly contribute to arylpiperazine ligands to form complexes with the DAR D2. Phe 178 (VI.44), Trp 182 (VI.48) and Tyr 216 (VII.58) can be considered as a part of the ancillary DAR D2 pocket preserved in most G protein coupled receptors of the A class and obviously, the arylpiperazine structural motif represents one of the privileged structures that bind to this pocket. PMID- 15819387 TI - Antidepressant and memory affecting influence of estrogen and venlafaxine in ovariectomized rats. AB - The experiments presented in this paper aimed to investigate whether estrogen level changes in oviariectomized rats (OVX) may lead to depression and memory disorders, and whether the effects of such changes may be reversible following administration of a new antidepressant, venlafaxine (CAS 9930-78-4, VEN, Efectin). The Porsolt forced swimming test and Morris water maze test were carried out on female Wistar rats after ovariectomy and in sham-ovariectomized rats. VEN 20 mg/kg was administered orally 30 min before the tests for the period of 14 days. Estradiol (17beta-estradiol benzoate, CAS 50-28-2, E2) administration (5 microg E2/0.2 ml sesame oil s.c.) was started 24 h after ovariectomy and was continued for 14 days--each dose was administered 180 min before the test. In the immobility test, which reflects antidepressant drug activity, it was found that VEN shortened immobility time (IT) after the 1st, 7th and 14th administration (days 1, 7, 14, respectively) in ovariectomized rats, whereas in the control group (sham-ovariectomized rats) VEN exerted antidepressant action only after single administration (day 1) and after 7 days of administration. E2 significantly reduced immobility behaviour both after single and chronic treatment in ovariectomized rats. After joint administration of VEN and E2 potentiation of the antidepressant activity of VEN could be observed in both groups except for concurrent administration of VEN and E2 after 14 days in sham ovariectomized rats. VEN improved the spatial memory in the Morris water maze test, whereas E2 did not affect the memory of the tested animals. Joint administration of VEN and E2 maintained the memory improving effect induced by VEN. The regulatory role of the steroid hormone and the new antidepressant drug (VEN) in antidepressant activity and memory function could be related to the interactions between noradrenergic and serotoninergic systems. PMID- 15819388 TI - Effects of a mononitrate, a beta1-blocker and a dihydropyridine calcium channel blocker on cardiovascular responsiveness to passive orthostasis: a placebo controlled double-blind study in normotensive volunteers. AB - OBJECTIVE: The aim of this study was to compare the influences of antianginal drugs such as mononitrate, beta-blocker and calcium channel blocker on cardiovascular responsiveness to orthostasis. METHODS: The responses to passive orthostasis (tilt provocation at 60 degrees for 3 min) were measured in normotensive healthy volunteers with whole-body impedance cardiography and finger blood-pressure monitoring after a single moderate oral dose of isosorbide-5 mononitrate (CAS 16051-77-7, 10 mg), the beta1-blocker bisoprolol fumarate (CAS 104344-23-2, 5 mg), the dihydropyridine calcium channel blocker nisoldipine (CAS 63675-72-9, 5 mg), and placebo in a randomised, double-blind fashion. RESULTS: In supine position, none of the drugs altered pre-tilt arterial pressure or heart rate (HR) when compared to placebo. Nisoldipine decreased systemic vascular resistance index (SVRI) when compared to either placebo or bisoprolol, and increased the cardiac index (CI) when compared to placebo. During the passive orthostasis, the mononitrate decreased SVRI when compared to placebo or bisoprolol. The mononitrate increased HR and pulse wave velocity (PWV) when compared to the other study groups, and decreased the stroke index when compared to placebo. In the bisoprolol group, the tilt responses of diastolic arterial pressure, HR, CI, left cardiac work index, and PWV decreased significantly compared to those in the placebo group. Nisoldipine did not alter the responses to orthostasis when compared to placebo. When compared to the mononitrate, both nisoldipine and bisoprolol decreased CI response to orthostasis. CONCLUSIONS: The mononitrate adversely affects the cardiovascular responsiveness to orthostasis. The beta-blocker reduces the responses and thus probably oxygen demand during orthostasis. The dihydropyridine calcium blocker seems to influence the responsiveness less than the mononitrate or beta-blocker. PMID- 15819389 TI - Effects of 15-deoxy-delta(12,14)-prostaglandin J2 on nuclear localization of GATA 3 in the murine lung in the presence of lipopolysaccharide. AB - 15-Deoxy-delta(12, 14)-prostaglandin J2 (15d-PG J2) is a regulator of a nuclear transcriptional factor, peroxisome proliferator-activated receptor (PPAR)-gamma. A previous study has demonstrated that 15d-PG J2 enhanced acute lung injury induced by lipopolysaccharide (LPS) in mice. 15d-PG J2 induced mucin-producing cells in the bronchial epithelium, especially in the presence of LPS. The present study investigated the effects of 15d-PG J2 on the activation of GATA-3 and Signal Transducer and Activator of Transcription (STAT) 6, important transcriptional factors in mucus secretion, in the lung in the presence or absence of LPS. ICR mice were divided into 4 experimental groups that intratracheally received vehicle, lipopolysaccharide (LPS: 125 microg/kg), 15d-PG J2 (1 mg/kg), or 15d-PG J2 + LPS. The nuclear localization of GATA-3 and phosphorylated STAT 6 was evaluated 2 h after the intratracheal administration. 15d-PG J2 enhanced the nuclear localization of GATA-3 in the presence of LPS, whereas the nuclear localization of phosphorylated STAT 6 was not altered in the groups. These results suggest that the enhancing effects of 15d-PG J2 on the production of mucin-producing cells might be related, at least in part, to the activation of GATA-3. PMID- 15819390 TI - Synthesis with improved yield and study on the analgesic effect of 2 hydroxyphencyclidine. AB - Phencyclidine (1-(1-phenylcyclohexyl)piperidine, CAS 956-90-1, PCP) has shown analgesic effects. Some of its derivatives have been synthesized and their biological properties were studied. Since a hydroxyl group has been added to the position 2 of the cyclohexane ring of PCP, this compound would be more hydrophilic than PCP. This compound was synthesized using a different and improved method with a higher yield. Its analgesic effect was studied using the tail-flick test on rats and was compared with that of ketamine (CAS 1867-66-9). The results showed that 2-hydroxyphencyclidine can increase tail-flick latencies as compared to the control group and indicate that the maximum analgesic effect of this compound occurs 2-5 min after its injection while the effect of ketamine is observed 10-25 min after injection. PMID- 15819391 TI - Selective toxicity of a quinoxaline 1,4-di-N-oxide derivative in human tumour cell lines. AB - The presence of hypoxic cells in human solid tumours is one of the causes of tumour resistance to conventional therapy, and is also associated with processes that promote the tumour progression. Different chemical agents have been designed in order to take advantage of the particular metabolic characteristics of hypoxic regions. These drugs, called bioreductive agents, are activated inside the hypoxic cells to give active species that, in the presence of oxygen, are oxidised back to the non-toxic parent compound. Several quinoxaline 1,4-di-N oxides have been described as potential bioreductive agents, and among them, 7 cloro-3-[[(N,N-dimethylamino)propy]amino]-2-quinoxalinecarbonitrile 1,4-di-N oxide hydrocloride (Q-85 HCl) appeared to be the most promising one. In the present work, the selective cytotoxicity of Q-85 HCl was studied in several human tumour cell lines of different origin (Caco2, MCF-7, HT-29 and Tk-10). Cell viability was calculated after 2 h treatment under hypoxic and well-oxygenated conditions. The potency (the concentration that gives 1% of cell survival) in hypoxia and hypoxia cytotoxicity ratio (HCR = potency in oxygenated conditions/potency in hypoxia) were calculated after a 14-day clonogenic assay. Q 85 HCl was more toxic in hypoxia than in well-oxygenated cells in all the tumour cell lines. The best profile of potency in hypoxia (0.4 micromol/L) and selectivity (HCR=155) was found in CaCo-2 cells. Altogether, these results suggest an in vitro biological profile for Q-85 HCl that makes it an interesting candidate for the development as a bioreductive agent. PMID- 15819392 TI - Visualizing genetic activity of mammalian cells: looking back to the year 1972. PMID- 15819393 TI - The retinoblastoma protein--from bench to bedside. AB - The retinoblastoma tumour suppressor protein (Rb) has come a long way since its initial discovery in 1986. Encoded by the first candidate tumour suppressor gene it has emerged a versatile and context-dependent modulator of cell behaviour. Its activity is managed by signalling networks sensing intra- and extracellular cues. These cues are relayed to hold or permit inactivation of Rb by phosphorylation. Loss or mutation of the retinoblastoma gene is rare in sporadic cancers but defects in the pathways that license inactivation of Rb are found in the majority of them, suggesting that loss of Rb control is central to tumour development and arguing that its reinstatement might reverse tumour formation. Furthermore, mouse models with engineered defects in the Rb-phosphorylating kinases provide evidence that moderation of Rb inactivation may be a strategy for the prevention of tumour formation. The rationale behind these arguments, their underlying molecular concepts and strategies towards therapeutic application will be discussed. PMID- 15819394 TI - Histone deacetylase inhibitors and cancer: from cell biology to the clinic. AB - Aberrant gene regulation plays an important role in tumor initiation and progression, and the acetylation of histones is a well understood key component of gene regulation. Histone acetylation involves the opposing activities of the histone acetyltransferases (HATs) and histone deacetylases (HDACs)--histone acetylation is associated with increased transcriptional activity while histone deacetylation is associated with repression of gene expression. In addition, the modification of non-histone proteins by HATs and HDACs is also an important process in regulating gene expression. Several lines of evidence suggest that inappropriate transcriptional activation and repression mediated by HATs and HDACs is a common occurrence in the formation of many different types of cancer. These enzymes thus represent novel molecular targets for which inhibitors are sought that could reprogram transcription and inhibit tumor cell growth and progression. Much of the research has focused on HDAC inhibitors, where several agents have demonstrated in vitro and in vivo activity against different tumor cell models and have entered Phase I clinical trials. HDAC inhibitors are believed to exert their antiproliferative effects by inducing a small set of genes involved in regulating cellular activities such as proliferation and differentiation. Future research is expected to lead to a better understanding of the molecular targets of HDACs and facilitate the development of more potent inhibitors of these enzymes. First results from clinical trials will help to determine the optimal strategy for utilizing these agents for the treatment of cancer patients. PMID- 15819395 TI - Chromatin, epigenetics and stem cells. AB - Epigenetics is a term that has changed its meaning with the increasing biological knowledge on developmental processes. However, its current application to stem cell biology is often imprecise and is conceptually problematic. This article addresses two different subjects, the definition of epigenetics and chromatin states of stem and differentiated cells. We describe mechanisms that regulate chromatin changes and provide an overview of chromatin states of stem and differentiated cells. Moreover, a modification of the current epigenetics definition is proposed that is not restricted by the heritability of gene expression throughout cell divisions and excludes translational gene expression control. PMID- 15819396 TI - Organization of the genome and gene expression in a nuclear environment lacking histones and nucleosomes: the amazing dinoflagellates. AB - Dinoflagellates are fascinating protists that have attracted researchers from different fields. The free-living species are major primary producers and the cause of harmful algal blooms sometimes associated with red tides. Dinoflagellates lack histones and nucleosomes and present a unique genome and chromosome organization, being considered the only living knockouts of histones. Their plastids contain genes organized in unigenic minicircles. Basic cell structure, biochemistry and molecular phylogeny place the dinoflagellates firmly among the eukaryotes. They have G1-S-G2-M cell cycles, repetitive sequences, ribosomal genes in tandem, nuclear matrix, snRNAs, and eukaryotic cytoplasm, whereas their nuclear DNA is different, from base composition to chromosome organization. They have a high G + C content, highly methylated and rare bases such as 5-hydroxymethyluracil (HOMeU), no TATA boxes, and form distinct interphasic dinochromosomes with a liquid crystalline organization of DNA, stabilized by metal cations and structural RNA. Without histones and with a protein:DNA mass ratio (1:10) lower than prokaryotes, they need a different way of packing their huge amounts of DNA into a functional chromatin. In spite of the high interest in the dinoflagellate system in genetics, molecular and cellular biology, their analysis until now has been very restricted. We review here the main achievements in the characterization of the genome, nucleus and chromosomes in this diversified phylum. The recent discovery of a eukaryotic structural and functional differentiation in the dinochromosomes and of the organization of gene expression in them, demonstrate that in spite of the secondary loss of histones, that produce a lack of nucleosomal and supranucleosomal chromatin organization, they keep a functional nuclear organization closer to eukaryotes than to prokaryotes. PMID- 15819397 TI - Nuclear envelopes from amphibian oocytes--from morphology to protein inventory. PMID- 15819398 TI - The nuclear lamina in Heidelberg and Wurzburg: a personal view. PMID- 15819399 TI - The mechanism of granulocyte nuclear shape determination: possible involvement of the centrosome. AB - Mature blood neutrophils (polymorphonuclear granulocytes) have characteristically complex nuclear shapes. The human neutrophil nucleus generally possesses 3-4 lobes; the mouse neutrophil nucleus frequently resembles a twisted toroid with a central hole. Myeloid tissue culture systems (e.g., human HL-60 and murine MPRO) can be induced to differentiate in vitro towards neutrophils by addition of retinoic acid, exhibiting the characteristic nuclear shape changes. Confocal immunostaining and thin-section transmission electron microscopic image data from differentiated HL-60 and MPRO cells clearly demonstrate proximity of the centrosomal region (containing dynein, gamma-tubulin and C-Nap1) to regions of granulocytic nuclear indentations. In addition, the centrosomal region, flanked by the Golgi apparatus, is shown to be present within the central hole of the toroidal mouse granulocyte nucleus. A role for the centrosomal region and associated microtubules in molding granulocytic nuclear shape is suggested. PMID- 15819400 TI - Plakophilins--hard work in the desmosome, recreation in the nucleus? AB - The linkage of the different types of cytoskeletal proteins to cell adhesion structures at the cytoplasmic membrane and the connection of these contact sites to corresponding sites of adjacent cells is a prerequisite for integrity and stability of cells and tissues. The structurally most prominent types of such cell-cell adhesion complexes are the desmosomes (maculae adhaerentes), which are found in all epithelia and certain non-epithelial tissues. As an element of the cytoskeleton, intermediate filaments are connected to the adhesive desmosomal transmembrane proteins by the cytoplasmic desmosomal plaque proteins. At least three different types of proteins are found in the desmosomal plaque, one of which is represented by the plakophilins, a recently described sub-family of sequence-related armadillo-repeat proteins. Consisting of three isoforms, plakophilins (plakophilin 1 to 3, PKP 1 to 3) are located in all desmosomes in a differentiation-dependent manner. While PKP 2 and PKP 3 are part of almost all desmosome-bearing cell types (PKP 2 except for differentiated cells of stratified epithelia and PKP 3 for hepatocytes and cardiomyocytes), PKP 1 is restricted to desmosomes of cells of stratified and complex epithelia. Besides the architectural function that plakophilins seem to fulfill in the desmosomes, at least PKP 1 and 2 are also localized in the nucleus independently of any differentiation-related processes and with an up to now enigmatic function in this compartment. In the following article we want to summarize the current knowledge concerning structure, function and regulation of the plakophilins that has been achieved during the last decade. PMID- 15819401 TI - The p120 family of cell adhesion molecules. AB - p120 is the prototypic member of the p120 subfamily of armadillo-related proteins that includes p0071, delta-catenin/NPRAP, ARVCF and the more distantly related plakophilins 1-3. Like armadillo, beta-catenin and plakoglobin these proteins are involved in mediating cell-cell adhesion. Besides their junctional localization they also reveal a cytoplasmic and nuclear localization. Non-cadherin-associated, cytoplasmic p120 functions in Rho signaling and regulation of cytoskeletal organization and actin dynamics. The nuclear function remains largely unsolved. Some characteristics seem to be shared by the various members of the family but it seems unlikely that p120-related proteins have solely redundant functions and compete for interactions with identical binding partners. Stabilization of cadherins at the membrane seems a common function of p120, p0071, delta-catenin and ARVCF but it is not yet known if and how these proteins confer distinct properties to cellular junctions. Moreover, p0071, NPRAP and ARVCF have a C terminal PDZ-binding motif that is lacking in p120 pointing to distinct roles of these proteins. PDZ domains are found in a series of proteins involved in establishing cell polarity in epithelial cells. Thus, p120 proteins may not only be master regulators of cadherin abundance and activity but play additional roles in regulating cell polarity. This review focuses on the putative roles of p120 proteins in cell polarity. PMID- 15819402 TI - Desmosomal cell adhesion in mammalian development. AB - Defects in desmosome-mediated cell-cell adhesion can lead to tissue fragility syndromes. Both inherited and acquired diseases caused by desmosomal defects have been described. The two organs that appear most vulnerable to these defects are the skin with its appendages, and the heart. Furthermore, the analysis of genetically engineered mice has led to the discovery that desmosomal proteins are also required for normal embryonic development. Knockout mice for several desmosomal proteins die in utero. Depending on the protein studied, death occurs either around the time of implantation, at mid-gestation or shortly before birth. So far, it appears that structural defects leading to abnormal histo-architecture and tissue fragility are the main cause of death, i.e. there is no evidence that loss of a desmosomal protein would abort specific cell lineages or differentiation programs. Nevertheless, we are only beginning to understand the functions of individual desmosomal proteins during development. This review focuses on the role of desmosomes during mouse embryonic development. PMID- 15819403 TI - Cadherin dimers in cell-cell adhesion. AB - While the critical function of classic cadherin in cell-cell junctions is well established, the molecular mechanism of cadherin-based adhesion remains unclear. The elusive but principal part of this adhesion process is the cadherin-cadherin interaction maintaining the intercellular contacts. This interaction is believed to be weak, suggesting that the adhesive contacts are strengthened by the cytoskeleton-dependent clustering of numerous cadherin molecules. An examination of cadherin homodimers in living cells has shown, however, that cadherin adhesive interaction is surprisingly strong. This observation implies that the strength of the adhesive contacts is regulated by the processes disintegrating cadherin dimers. The molecular structure of these dimers and mechanisms potentially responsible for their dynamics in living cells are discussed in this review. PMID- 15819404 TI - Signaling from adherens-type junctions. PMID- 15819405 TI - Intracellular origin and secretion of milk fat globules. AB - The cream or fat fraction of milk consists of fat droplets composed primarily of triacylglycerols that are surrounded by cellular membranes. In this review we discuss what is known about how these droplets are formed in and secreted by mammary epithelial cells during lactation. This secretion mechanism, which appears to be unique, is unlike the exocytotic mechanism used by other cell types to secrete lipids. Milk fat globules originate as small, triacylglycerol-rich, droplets that are formed on or in endoplasmic reticulum membranes. These droplets are released from endoplasmic reticulum into the cytosol as microlipid droplets coated by proteins and polar lipids. Microlipid droplets can fuse with each other to form larger cytoplasmic lipid droplets. Droplets of all sizes appear to be unidirectionally transported to apical cell regions by as yet unknown mechanisms that may involve cytoskeletal elements. These lipid droplets appear to be secreted from the cell in which they were formed by being progressively enveloped in differentiated regions of apical plasma membrane. While plasma membrane envelopment appears to be the primary mechanism by which lipid droplets are released from the cell, a mechanism involving exocytosis of lipid droplets from cytoplasmic vacuoles also has been described. As discussed herein, while we have a general overview of the steps leading to the fat globules of milk, virtually nothing is known about the molecular mechanisms involved in milk fat globule formation, intracellular transit, and secretion. PMID- 15819406 TI - Human Merkel cells--aspects of cell biology, distribution and functions. AB - Human Merkel cells were first described by Friedrich S. Merkel in 1875 and named "Tastzellen" (touch cells) assuming a sensory touch function within the skin. Only ultrastructural research revealed their characteristics such as dense-core granules, plasma membrane spines and dendrites as well as a loosely arranged cytoskeleton. Biochemical analysis identified the expression of very specific cytokeratins (most notably CK 20) allowing the immunohistochemical detection of Merkel cells. In humans, they occur within the basal epidermis, being concentrated in eccrine glandular ridges of glabrous skin and in Haarscheiben of hairy skin, within belt-like clusters of hair follicles, and in certain mucosal tissues. Within the human skin, the dense-core granules contain heterogeneously distributed neuropeptides, some of which might work as neurotransmitters through which Merkel cells and their associated nerves exert their classical function as slowly adapting mechanoreceptors type I. This is the case in the Haarscheiben, small sensory organs containing keratinocytes with a special program of differentiation that includes the expression of CK 17 and Ber-EP4. Other peptides may act as growth factors and thus might participate in growth, differentiation and homeostasis of cutaneous structures. It is not yet clear whether the Merkel cell carcinomas, aggressive skin carcinomas, indeed arise from Merkel cells. We summarize and discuss data on the distribution, function and heterogeneity of human Merkel cells in normal and diseased skin. PMID- 15819407 TI - Immune dysregulation in lichen sclerosus. AB - Lichen sclerosus (LS) is a chronic localized lymphocyte-mediated dermatosis of genital skin with a presumed autoimmune origin. LS is characterized by localized dense lymphocytic tissue infiltrates, vasculitic processes and extensive tissue destruction. The lymphocytic infiltrate of LS biopsies contains between 1.4% and 21% of T-cells with monoclonally rearranged T-cell receptor gamma-chain gene, and the immunophenotype is dominated by B-cells, CD4-positive T-cells and antigen presenting dendritic cells. Antigen-driven selection of T-cells and restricted T cell receptor usage reflects prolonged exposure of the host immune system to a local (putative LS-associated) antigen. It is presently unclear at which time point in the evolution of LS the T-cell clones emerge. All investigators of LS agree on the non-neoplastic nature of the infiltrate. However, a small percentage of LS patients show serological (systemic) evidence of T-cell immune deficiencies. The lack of long-term follow up of patients with known monoclonally rearranged T-cell receptor gamma-chain gene in their LS biopsies, however, defers a final judgement on the clinical significance of our observations. PMID- 15819408 TI - Protein NO52--a constitutive nucleolar component sharing high sequence homologies to protein NO66. AB - The nucleolus is the most prominent intranuclear structure of almost all protein synthesizing cells. It compromises a well-defined functional compartmentalization and a high complexity of molecular constituents. Here, we report on the identification and molecular characterization of a novel constitutive nucleolar component--protein NO52--that is present in diverse species from Xenopus laevis to human. The cDNA-deduced amino acid sequence of protein NO52 defines a polypeptide of a calculated mass of 52.8 kDa and an isoelectric point of 6.7. Inspection of the primary sequence disclosed that the protein contains a JmjC domain and is highly sequence-related to the recently described nucleolar protein NO66. Immunolocalization studies revealed that protein NO52 is highly concentrated in the granular component of nucleoli and this characteristic intranuclear distribution is significantly affected by treatment of cells with (i) RNase A, (ii) actinomycin D and (iii) serum starvation. Interestingly, protein NO52 has been identified as a constituent of free preribosomal particles but is absent from cytoplasmic ribosomes. Analyses of immunocomplexes isolated from cellular extracts with an NO52-specific antibody by MALDI mass spectrometry further confirmed the interaction of protein NO52 with various ribosomal proteins as well as with a distinct set of non-ribosomal nucleolar proteins. The dependence of the nucleolar accumulation of the protein on ongoing rRNA transcription and the cellular metabolic state strongly suggest that protein NO52 is directly involved in ribosome biogenesis, most likely during the assembly process of preribosomal particles. PMID- 15819409 TI - Emerin expression in early development of Xenopus laevis. AB - Emerin is an integral protein of the inner nuclear membrane in the majority of differentiated vertebrate cells. In humans, deficiency of emerin causes a progressive muscular dystrophy of the Emery-Dreifuss type. The physiological role of emerin is poorly understood. By screening and sequencing of EST clones we have identified two emerin homologues in Xenopus laevis, Xemerin1 and Xemerin2. Xemerins share with mammalian emerins the N-terminal LEM domain and a single transmembrane domain at the C-terminus. As shown by immunoblot analysis with Xemerin-specific antibodies, both proteins have an apparent molecular mass of 24 kDa but differ in their isoelectric points. Xemerin1 and Xemerin2 proteins are not detectable in oocytes nor during early embryogenesis. Protein expression is first found at stage 43 and persists in somatic cells. However, RT-PCR and Northern blot analysis show Xemerin mRNAs of approximately 4.0 kb to be present in oocytes and throughout embryogenesis. During embryogenesis the level of Xemerin mRNAs increases at stage 22 and is particularly abundant in mesodermal and neuro-ectodermal regions of the embryo. These data provide the necessary background to further investigate the role of emerin in nuclear envelope assembly, gene expression and organ development of X. laevis as a model organism. PMID- 15819410 TI - Dissection of keratin dynamics: different contributions of the actin and microtubule systems. AB - It has only recently been recognized that intermediate filaments (IFs) and their assembly intermediates are highly motile cytoskeletal components with cell-type- and isotype-specific characteristics. To elucidate the cell-type-independent contribution of actin filaments and microtubules to these motile properties, fluorescent epithelial IF keratin polypeptides were introduced into non epithelial, adrenal cortex-derived SW13 cells. Time-lapse fluorescence microscopy of stably transfected SW13 cell lines synthesizing fluorescent human keratin 8 and 18 chimeras HK8-CFP and HK18-YFP revealed extended filament networks that are entirely composed of transgene products and exhibit the same dynamic features as keratin systems in epithelial cells. Detailed analyses identified two distinct types of keratin motility: (I) Slow (approximately 0.23 microm/min), inward directed, continuous transport of keratin filament precursor particles from the plasma membrane towards the cell interior, which is most pronounced in lamellipodia. (II) Fast (approximately 17 microm/min), bidirectional and intermittent transport of keratin particles in axonal-type cell processes. Disruption of actin filaments inhibited type I motility while type II motility remained. Conversely, microtubule disruption inhibited transport mode II while mode I continued. Combining the two treatments resulted in a complete block of keratin motility. We therefore conclude that keratin motility relies both on intact actin filaments and microtubules and is not dependent on epithelium specific cellular factors. PMID- 15819411 TI - Interaction of stress proteins with misfolded keratins. AB - Misfolded and aggregated proteins are a characteristic feature of a variety of chronic diseases. Examples include neurofibrillary tangles in Alzheimer disease, Lewy bodies in Parkinson disease and Mallory bodies (MBs) in chronic liver diseases, particularly alcoholic and non-alcoholic steatohepatitis (ASH and NASH). MB formation is at least in part the result of chronic oxidative cell stress in hepatocytes and can be induced in mice by long-term intoxication with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC). Proteomic analysis revealed that MBs consist of ubiquitinated keratins and the stress proteins Hsp70, Hsp25, and p62. Furthermore, marked overexpression of clusterin, which shares functional properties with small heat shock proteins, was identified by gene expression profiling of DDC-treated mice livers. To investigate whether clusterin has a function in the stress response to misfolded keratins, we performed transfection studies utilizing expression constructs encoding ubiquitin, p62, Hsp27, clusterin, keratin 8, and keratin 18. Ubiquitin was found in a strong and constant association with keratin aggregates, whereas binding of p62 to keratin was variable. Hsp27 did not colocalize with keratin aggregates under these experimental conditions. In contrast, clusterin associated with misfolded keratin only if its signal peptide was deleted and its secretion inhibited. This suggests that clusterin has ability to bind misfolded proteins, including keratins but its physiological function is restricted to the extracellular space. The extracellular localization of clusterin was underlined by immunohistochemical studies in Alzheimer disease brains, where clusterin was constantly found in association with amyloid plaques; in contrast, cytoplasmic inclusions such as neurofibrillary tangles as well as MBs in ASH were negative. Furthermore, we found clusterin in association with elastic fibers in the extracellular matrix in several chronic liver diseases, including ASH and alpha1-antitrypsin deficiency, implying a possible role of clusterin in liver fibrosis. PMID- 15819413 TI - Rescue of keratin 18/19 doubly deficient mice using aggregation with tetraploid embryos. AB - We have previously shown that the targeted deletions of both type I keratins (K) 18 and 19 cause lethality by embryonic day (e) 9.5 due to fragility and cytolysis of trophoblast giant cells. The development of the embryo proper appeared to be unaffected and its death was caused by nutrient deficiency. In order to address the function of keratins within the embryo proper, lethality due to extraembryonic tissue failure must be overcome. One approach to rescue doubly deficient embryos is by aggregating knockout embryos with tetraploid wild-type embryos. As a general tool, tetraploid aggregation can be used to rescue embryonic lethality caused by defects in extraembryonic tissues like the placenta, trophoblast or yolk sac. We rescued K18-/- K19-/- embryos until e11.5, using this approach, proving that the loss of the keratin cytoskeleton causes defects in the trophoblast giant cell layer, but has no effect on early development of the embryo proper. PMID- 15819412 TI - Identification of a stem cell candidate in the normal human prostate gland. AB - Stem cells of the human prostate gland have not yet been identified utilizing a structural biomarker. We have discovered a new prostatic epithelial cell phenotype-expressing cytokeratin 6a (Ck6a+ cells). The Ck6a+ cells are present within a specialized niche in the basal cell compartment in fetal, juvenile and adult prostate tissue, and within the stem cell-enriched urogenital sinus. In adult normal prostate tissue, the average abundance of Ck6a+ cells was 4.9%. With proliferative stimuli in the prostate organ culture model, in which the epithelial-stromal interaction was maintained, a remarkable increase of Ck6a expression was noticed to up to 64.9%. The difference in cytokeratin 6a expression between the normal adult prostate and the prostate organ culture model was statistically significant (p<0.0001). Within the prostate organ culture model the increase of cytokeratin 6a-expressing cells significantly correlated with increased proliferation index (r = 0.7616, p = 0.0467). The Ck6a+ cells were capable of differentiation as indicated by their expression of luminal cell markers such as ZO-1 and prostate specific antigen (PSA). Our data indicate that Ck6a+ cells represent a prostatic epithelial stem cell candidate possessing high potential for proliferation and differentiation. Since the development of benign prostatic hyperplasia and prostate carcinogenesis are disorders of proliferation and differentiation, the Ck6a+ cells may represent a major element in the development of these diseases. PMID- 15819414 TI - Evolution of tissue-specific keratins as deduced from novel cDNA sequences of the lungfish Protopterus aethiopicus. AB - Lungfishes are possibly the closest extant relatives of the land vertebrates (tetrapods). We report here the cDNA and predicted amino acid sequences of 13 different keratins (ten type I and three type II) of the lungfish Protopterus aethiopicus. These keratins include the orthologs of human K8 and K18. The lungfish keratins were also identified in tissue extracts using two-dimensional polyacrylamide gel electrophoresis, keratin blot binding assays and immunoblotting. The identified keratin spots were analyzed by peptide mass fingerprinting which assigned seven sequences (inclusively Protopterus K8 and K18) to their respective protein spot. The peptide mass fingerprints also revealed the fact that the major epidermal type I and type II keratins of this lungfish have not yet been sequenced. Nevertheless, phylogenetic trees constructed from multiple sequence alignments of keratins from lungfish and distantly related vertebrates such as lamprey, shark, trout, frog, and human reveal new insights into the evolution of K8 and K18, and unravel a variety of independent keratin radiation events. PMID- 15819415 TI - Characterization of the in vitro co-assembly process of the intermediate filament proteins vimentin and desmin: mixed polymers at all stages of assembly. AB - We have investigated the co-assembly properties of the intermediate filament (IF) proteins vimentin and desmin. First, the soluble complexes formed by both proteins separately in 5 mM Tris-HCl, pH 8.4, were characterized by analytical ultracentrifugation. In both cases, s-values of around 5 S were obtained corresponding to the formation of tetramers. However, at pH 7.5 and in the presence of 1 mM EDTA, both proteins behaved quite differently; whereas vimentin sedimented at 7.2 S, desmin assembled into much larger complexes of about 13 S. A mixture of equimolar amounts of vimentin and desmin in 8 M urea yielded, after reconstitution into 5 mM Tris-HCl, pH 7.5, and 1 mM EDTA, complexes exhibiting a sharp peak at 10.9 S. This intermediate s-value indicated that co-assembly into a distinct new set of complexes had occurred. As judged by electron microscopy and viscometry, these mixtures assembled into IFs with characteristics similar to those of pure vimentin and desmin. Furthermore, when vimentin and desmin tetramers were mixed in 5 mM Tris-HCl, pH 8.4, and subsequently subjected to IF assembly conditions, again "hybrid" filaments were obtained. Most interestingly, after 10 min of assembly, mass-per-length (MPL) measurements by scanning transmission electron microscopy yielded IFs with an MPL-peak value of 36 +/- 5 kDa/nm, hence closer to that of vimentin IFs (33 +/- 4 kDa/nm) than to that of desmin IFs (48 +/- 8 kDa/nm). Finally, when unit length-filaments (ULF) of vimentin and desmin were mixed and assembled further, the diameters of individual mature IFs formed exhibited a significantly higher degree of width inhomogeneity along their length than vimentin and desmin IFs as might be expected for a modular mode of assembly. Last but not least, atomic force microscopy provided further direct evidence that desmin IFs are able to fuse end-to-end with vimentin IFs. In summary, we have shown that vimentin and desmin are able to co-assemble at the dimer, tetramer, ULF and even the mature IF level. PMID- 15819416 TI - Urothelial umbrella cells of human ureter are heterogeneous with respect to their uroplakin composition: different degrees of urothelial maturity in ureter and bladder? AB - Urothelial umbrella cells are characterized by apical, rigid membrane plaques, which contain four major uroplakin proteins (UP Ia, Ib, II and III) forming UPIa/UPII and UPIb/UPIII pairs. These integral membrane proteins are thought to play an important role in maintaining the physical integrity and the permeability barrier function of the urothelium. We asked whether the four uroplakins always coexpress in the entire human lower urinary tract. We stained immunohistochemically (ABC-peroxidase method) paraffin sections of normal human ureter (n = 18) and urinary bladder (n = 10) using rabbit antibodies against UPIa, UPIb, UPII and UPIII; a recently raised mouse monoclonal antibody (MAb), AU1, and two new MAbs, AU2 and AU3, all against UPIII; and mouse MAbs against umbrella cell-associated cytokeratins CK18 and CK20. Immunoblotting showed that AU1, AU2 and AU3 antibodies all recognized the N-terminal extracellular domain of bovine UPIII. By immunohistochemistry, we found that in 15/18 cases of human ureter, but in only 2/10 cases of bladder, groups of normal-looking, CK18 positive umbrella cells lacked both UPIII and UPIb immunostaining. The UPIb/UPIII negative cells showed either normal or reduced amounts of UPIa and UPII staining. These data were confirmed by double immunofluorescence microscopy. The distribution of the UPIb/UPIII-negative umbrella cells was not correlated with localized urothelial proliferation (Ki-67 staining) or with the distribution pattern of CK20. Similar heterogeneities were observed in bovine but not in mouse ureter. We provide the first evidence that urothelial umbrella cells are heterogeneous as some normal-looking umbrella cells can possess only one, instead of two, uroplakin pairs. This heterogeneity seems more prominent in the urothelium of human ureter than that of bladder. This finding may indicate that ureter urothelium is intrinsically different from bladder urothelium. Alternatively, a single lineage of urothelium may exhibit different phenotypes resulting from extrinsic modulations due to distinct mesenchymal influence and different degrees of pressure and stretch in bladder versus ureter. Additional studies are needed to distinguish these two possibilities and to elucidate the physiological and pathological significance of the observed urothelial and uroplakin heterogeneity. PMID- 15819417 TI - Both type-I hemidesmosomes and adherens-type junctions contribute to the cell substratum adhesion system in myoepithelial cells. AB - Myoepithelial cells present in exocrine glands cause secretion from the glands by contraction. They have mixed characteristics with regard to cytoskeletal elements, containing both epithelial-type intermediate filaments and smooth muscle-type myofilaments. For further characterization, myoepithelial cells from bovine apocrine sweat glands and tracheal glands were here examined with special attention to the cell-substratum adhesion system. Immunofluorescence microscopy using a panel of antibodies against adherens-type junctional and hemidesmosomal proteins demonstrated two types of cell-substratum junctions in myoepithelial cells from both glands. Type-I hemidesmosomes (HDs) consisting of plectin, BP230, integrin alpha6beta4, and BP180 were thus observed as punctate arrays longitudinally arranged along myoepithelial cell surfaces, while adherens-type junctions were similarly evident as linear rib-like structures. Double-label immunofluoresence revealed the two junctions to be distributed in a mutually exclusive or independent manner. Electron microscopy further demonstrated that apocrine myoepithelial cells surround secretory epithelial cells completely, without any gaps, HDs being abundant along the basement membrane, but with no distinct structures in the inter-hemidesmosomal regions. Immunoelectron microscopy, however, revealed an interhemidesmosomal localization of vinculin, pointing to the existence of adherens-type junctions. Secretory epithelial cells in tracheal glands were found not to be completely covered with myoepithelial cells, so that more than half of them are directly attached to the basement membrane, where they form type II-HDs lacking BP230 and BP180, but no detectable adherens junctions, like epidermal basal cells and sebaceous gland cells. These observations demonstrate that, in addition to their cytoskeleton, myoepithelial cells have both epithelial- and smooth muscle-type cell-substratum adhesion structures, i.e. HDs and dense plaque-like adherens junctions. PMID- 15819418 TI - Synthesis of junctional proteins in metastasizing colon cancer cells. AB - Various authors have reported reduced synthesis of epithelial junctional proteins during dedifferentiation, tumorigenesis and metastasis in a great variety of tumors. Consequently, it is generally accepted that loss of adhesive molecules and adhesion structures is implicated in the development of an invasive phenotype and poor patient prognosis. Colon carcinomas, on the other hand, were shown to behave differently as synthesis of main adhesive proteins continues despite the development of an invasive phenotype. In this study we used cultured cells grown under conditions that inhibited intercellular adhesion (low Ca2+ concentration) and compared these results with data obtained from metastasizing colon cancer cells (signet ring cell carcinoma). Characterization of these proteins and their structures were performed by immunoprecipitations, Western blot analysis, immunohistochemistry, pre-embedding immuno-electron microscopy, and a new method to perform immuno-electron microscopy on paraffin-embedded material, which we present in this paper. We demonstrate that synthesis carries on for both, the desmosomal and the proteins of the zonula adhaerens. While, however, the assembly of desmosomal structures in the form of half-desmosomes at the cell surface continues, those of the zonula adhaerens did not. Instead E-cadherin was found, although associated with alpha-catenin, beta-catenin, and plakoglobin, evenly distributed at the plasma membrane of the cultured cells and also at the surface of the dissociated tumor cells. We conclude from our observations that continued expression and synthesis of junctional proteins do not necessarily contribute to the suppression of tumor invasion and metastasis of colon cancer. PMID- 15819419 TI - Transcript and proteome analysis reveals reduced expression of calgranulins in head and neck squamous cell carcinoma. AB - The calcium-binding proteins of the S100 and the annexin protein families have been implicated in a variety of important physiological functions including membrane remodeling, calcium-related intracellular signaling, cytoskeleton dynamics, tissue homeostasis, and formation of the cornified envelope in differentiating keratinocytes. Deregulated expression of members of these families has been reported in different types of neoplasia and other diseases, but the results were not consistent. Here we have applied a combination of cDNA microarrays, quantitative reverse transcriptase-PCR (qRT-PCR) and surface enhanced laser desorption ionisation-time of flight mass spectrometry (SELDI-TOF MS) to study differential expression of these genes in head and neck squamous cell carcinoma (HNSCC). The calgranulins A and B and annexins 1 and 2 were found to be down-regulated in HNSCC, compared with normal mucosa, at both the mRNA and protein level. Upon validation of the differential gene expression by tissue microarray immunohistochemistry, we detected novel expression patterns of calgranulins A and B both in normal mucosa as well as in HNSCC. In contrast to squamous cancer of skin and other cancers in which the calgranulins were found to be up-regulated, most HNSCC showed reduced and widely deranged staining patterns including heterogeneous nuclear, cytoplasmic and membranous staining, and even enhanced staining in the tumor stroma. These observations suggest that the normal function of the calgranulins A and B in mucosa might be different from that in skin. PMID- 15819420 TI - The cancer/testis antigen CAGE-1 is a component of the acrosome of spermatids and spermatozoa. AB - Cancer/testis antigens (CTAs) are characterized by their restricted expression pattern. In normal individuals their expression is largely restricted to the testis. In the case of cancer patients, CTA expression has also been frequently observed in the tumoral cells. CTAs are considered to be promising targets for immunotherapy. However, almost nothing is known about the properties defined by the vast majority of CTAs. Here, we have investigated the expression pattern and localization of the CTA CAGE-1 during mouse spermatogenesis. We show that protein CAGE-1 is 849 amino acids long. Analysis of the first spermatogenic wave of pubertal mice by RT-PCR and immunoblotting showed that CAGE-1 is predominantly expressed during postmeiotic stages. CAGE-1 localizes to the acrosomal matrix and acrosomal granule, as demonstrated by immunocytochemistry at the light and electron microscopic level. Taken together, our results allowed to define protein CAGE-1 as a novel component of the acrosome of mammalian spermatids and spermatozoa. PMID- 15819421 TI - TuJ1 (class III beta-tubulin) expression suggests dynamic redistribution of follicular dendritic cells in lymphoid tissue. AB - Follicular dendritic cells (FDCs) play central roles in the B cell survival, proliferation, and differentiation into memory cells. Here, we show that TuJ1 (class III beta-tubulin) is expressed strongly in FDCs of human lymphoid tissue. TuJ1 has been a marker of neurons in the central and peripheral nervous systems from the early stage of neural differentiation. FDCs expressed TuJ1 protein diffusely in both light and dark zones of germinal centers in all human lymphoid tissues. In contrast, CD21 expression was relatively concentrated to the light zone, suggesting that TuJ1 was a marker for FDCs with broader spectrum than CD21. In addition to the germinal center, there were single TuJ1-expressing cells scattered in the mantle zone, blurring the border of the FDC network. In human tonsils, single scattered TuJ1-positive cells were also present in the crypt epithelium, suggesting a dynamic redistribution of FDCs among the antigen-rich epithelium, mantle zone, and germinal center. Such migration of FDCs could reflect a way of direct transport of various antigens carried on their surface to the germinal center, and a basis for the polarity of lymphoid follicles toward the epithelium in mucosa-associated lymphoid tissues. HK cells, cultured FDCs, also expressed TuJ1. The expression of TuJ1 by FDCs suggests that they may share certain biological characteristics of the neural system. PMID- 15819422 TI - From science to application. PMID- 15819423 TI - Influence of temperature on Ranavirus infection in larval salamanders Ambystoma tigrinum. AB - Temperature strongly influenced percent mortality and time to death of salamanders exposed to the Ambystoma tigrinum virus (iridovirus) (ATV). Most salamanders survived when exposed at 26 degrees C, whereas all died at 18 degrees C and nearly all died at 10 degrees C. Some asymptomatic salamanders that survived 60 d at 10 or 26 degrees C were found to be carrying virus. Polymerase chain reaction (PCR) confirmed the presence of virus in ATV-exposed salamanders but was found to be less sensitive than cell culture in detecting ATV at low concentrations. PCR products were 100% identical to ATV in the major capsid protein sequence. Virus titer was higher in salamanders held at 10 degrees C than at 18 degrees C but little virus, if any, was present in the small number of salamanders that died at 26 degrees C. These results may help explain periodic viral epizootics in field populations of A. tigrinum where water temperatures fluctuate widely. PMID- 15819424 TI - Taura syndrome virus (TSV) in Thailand and its relationship to TSV in China and the Americas. AB - The cultivation of exotic Penaeus vannamei in Thailand began on a very limited scale in the late 1990s, but a Thai government ban on the cultivation of P. monodon in freshwater areas in 2000 led many Thai shrimp farmers to shift to cultivation of P. vannamei. Alarmed by the possibility of Taura syndrome virus (TSV) introduction, the Thai Department of Fisheries required that imported stocks of P. vannamei be certified free of TSV by RT-PCR (Reverse Trasciption Polymerase Chain Reaction) testing. During the interval of allowed importation, over 150,000 broodstock shrimp were imported, 67% of these from China and Taiwan. Despite the safeguards, TSV outbreaks occurred and we confirmed the first outbreak by RT-PCR in early 2003. This resulted in a governmental ban on all shrimp broodstock imports from February 2003, but TSV outbreaks have continued, possibly due to original introductions or to the continued illegal importation of stocks. To determine the origin of the TSV in Thailand, the viral coat protein gene VP1 was amplified by RT-PCR from several shrimp specimens found positive for TSV by RT-PCR from January to November 2003. These included 7 samples from P. vannamei disease outbreaks in Thailand, 3 other non-diseased shrimp samples from Thailand and Burma and 6 samples including P. vannamei and P. japonicus from China. Comparison revealed that the Thai, Burmese and Chinese TSV types formed a clade distinct from a clade of TSV types from the Americas. PMID- 15819425 TI - Polychaete worms--a vector for white spot syndrome virus (WSSV). AB - The present work provides the first evidence of polychaete worms as passive vectors of white spot syndrome virus (WSSV) in the transmission of white spot disease to Penaeus monodon broodstocks. The study was based on live polychaete worms, Marphysa spp., obtained from worm suppliers/worm fishers as well as samples collected from 8 stations on the northern coast of Tamilnadu (India). Tiger shrimp Penaeus monodon broodstock with undeveloped ovaries were experimentally infected with WSSV by feeding with polychaete worms exposed to WSSV. Fifty percent of polychaete worms obtained from worm suppliers were found to be WSSV positive by 2-step PCR, indicating high prevalence of WSSV in the live polychaetes used as broodstock feed by hatcheries in this area. Of 8 stations surveyed, 5 had WSSV positive worms with prevalence ranging from 16.7 to 75%. Polychaetes collected from areas near shrimp farms showed a higher level of contamination. Laboratory challenge experiments confirmed the field observations, and > 60% of worms exposed to WSSV inoculum were proved to be WSSV positive after a 7 d exposure. It was also confirmed that P. monodon broodstock could be infected with WSSV by feeding on WSSV contaminated polychaete worms. Though the present study indicates only a low level infectivity in wild polychaetes, laboratory experiments clearly indicated the possibility of WSSV transfer from the live feed to shrimp broodstock, suggesting that polychaete worms could play a role in the epizootiology of WSSV. PMID- 15819426 TI - Nodavirus infection causes mortalities in hatchery produced larvae of Lates calcarifer: first report from India. AB - Larvae (15 to 21 d post hatch, dph) of the Asian sea bass Lates calcarifer (Bloch) suffered heavy mortalities (60 to 90%) during the hatchery-rearing phase. Darkened and moribund larvae showed no evidence of bacterial or parasitic infections. Tissue sections of brain and spinal cord showed clear necrotic vacuolation. Electron microscopy revealed membrane-bound viral particles in the cytoplasm of the nerve cells. The viral particles measured 28 to 30 nm in diameter. Primer sets, designed for the amplification of the RNA2 segment of the piscine nodavirus coat protein gene, were used in the RT-PCR analysis of moribund larvae of 20 and 21 dph which produced the amplified product of 430 bp. The clinical manifestations, pathology and electron microscopy observations supported by the RT-PCR analysis suggest that the nerve necrosis was due to nodavirus infection in the larvae. This is the first report of piscine nodavirus infection from the Indian sub-continent. PMID- 15819427 TI - Application of surveillance data in evaluation of diagnostic tests for infectious salmon anemia. AB - Infectious salmon anemia (ISA) is a viral disease in farmed Atlantic salmon Salmo salar, characterized by lethargy, anorexia, anemia and death. Test methods used for regulatory decisions to remove infected cages include the indirect fluorescent antibody test (IFAT), the reverse transcription-polymerase chain reaction test (RT-PCR), and virus isolation (VI). However, no thorough evaluation of these diagnostic tests has been carried out on field samples. The objective of this study was to evaluate the sensitivity and specificity of ISA diagnostic tests as individual tests and in combinations, using data collected by the provincial government surveillance program. Because a 'gold standard' reference test for ISA was not available, cage status was based on clinical disease records. A pool of fish from negative farms that had never had clinical ISA and a pool of fish from positive cages that were experiencing an outbreak of clinical ISA were obtained and assumed to be uninfected and infected respectively. A total of 1071 fish were used in this study. Depending on the test's cut-off value, the sensitivity and specificity for histopathology ranged from 30 to 73 and 72 to 99% respectively. IFAT had sensitivities and specificities in the range of 64 to 83 and 96 to 100% respectively. For the RT-PCR, sensitivity and specificity were 93 and 98% respectively. Test performances were also evaluated in series and in parallel combinations. Sensitivities are maximized when tests are evaluated in parallel, and ranged from 75 to 98%. Specificities are maximized when the tests are evaluated in series, and ranged from 99 to 100%. Current surveillance testing protocols should be reviewed to capitalize on this newly available information on test characteristics. PMID- 15819428 TI - Transmission and detection of Flavobacterium columnare in channel catfish Ictalurus punctatus. AB - A specific and rapid PCR detection method for Flavobacterium columnare based on the 16S-23S rDNA intergenic spacer region (ISR) of the ribosomal RNA operon has been developed. The ISR of 30 F. columnare strains and other Flavobacterium species was amplified using universal primers and sequenced. Once F. columnare specific sequences within the ISR were recognized, specific PCR primers were designed against them (FCISRFL and FCISRR1). The primers were sensitive and able to detect as low as 7 colony forming units from pure culture by PCR. The new PCR detection method was applied to experimentally infected channel catfish. Two different experiments in which channel catfish fingerlings were infected by intramuscular injection or by immersion bath showed the advantage of the PCR method over standard culture techniques. F. columnare was detected by PCR in both tank water and catfish tissue samples with a higher frequency and in less time than standard microbiological methods. Furthermore, PCR detection confirmed that F. columnare can be transmitted horizontally indirectly through the water column without fish-to-fish contact. The newly developed PCR detection method for F. columnare was more sensitive and rapid than standard culture on bacteriological media for detection of F. columnare in channel catfish tissues and in tank water. PMID- 15819429 TI - Genetic characterization and experimental pathogenesis of Piscirickettsia salmonis isolated from white seabass Atractoscion nobilis. AB - An intracellular bacterium originally isolated from hatchery-reared juvenile white seabass Atractoscion nobilis in southern California, USA, was identified by sequences of the small and large subunit ribosomal (16S and 23S) DNA and the internal transcribed spacer (ITS) as Piscirickettsia salmonis. Considering all rDNA sequences compared, the white seabass isolate (WSB-98) had a 96.3 to 98.7% homology with 4 previously described strains of P. salmonis isolated from salmon in Chile, Norway, and British Columbia, Canada. Experimental infections induced by intraperitoneal injections of juvenile white seabass with WSB-98 resulted in disease and mortality similar to that observed in P. salmonis infections in salmon. After 60 d, the cumulative mortality among P. salmonis-injected white seabass was 82 and 40%, respectively, following a high (1.99 x 10(4) TCID50) or low (3.98 x 10(2) TCID50) dose-challenge with WSB-98. The bacterium was recovered by isolation in cell culture or was observed in stains from tissues of injected white seabass but not from control fish. There were no external signs of infection. Internally, the most common gross lesion was a mottled appearance of the liver, sometimes with distinct nodules. Microscopic lesions were evident in both the capsule and parenchyma of the liver and were characterized by multifocal necrosis, often with infiltration of mononuclear leukocytes. Macrophages filled with bacteria were present at tissue sites exhibiting focal necrosis. Foreign body-type granulomas were prevalent in livers of experimentally infected white seabass, but not in control fish. Similar granulomatous lesions were observed in the spleen, kidney, intestine and gills, but these organs were considered secondary sites of infection, with significantly fewer and less severe histologic lesions compared to the liver. The results from this study clearly indicate that infections with P. salmonis are not restricted to salmonid fishes and that the bacterium can cause a disease similar to piscirickettsiosis in nonsalmonid hosts. PMID- 15819430 TI - A cohabitation challenge to compare the efficacies of vaccines for bacterial kidney disease (BKD) in chinook salmon Oncorhynchus tshawytscha. AB - The relative efficacies of 1 commercial and 5 experimental vaccines for bacterial kidney disease (BKD) were compared through a cohabitation waterborne challenge. Groups of juvenile chinook salmon Oncorhynchus tshawytscha were vaccinated with one of the following: (1) killed Renibacterium salmoninarum ATCC 33209 (Rs 33209) cells; (2) killed Rs 33209 cells which had been heated to 37 degrees C for 48 h, a process that destroys the p57 protein; (3) killed R. salmoninarum MT239 (Rs MT239) cells; (4) heated Rs MT239 cells; (5) a recombinant version of the p57 protein (r-p57) emulsified in Freund's incomplete adjuvant (FIA); (6) the commercial BKD vaccine Renogen; (7) phosphate-buffered saline (PBS) emulsified with an equal volume of FIA; or (8) PBS alone. Following injection, each fish was marked with a subcutaneous fluorescent latex tag denoting its treatment group and the vaccinated fish were combined into sham and disease challenge tanks. Two weeks after these fish were vaccinated, separate groups of fish were injected with either PBS or live R. salmoninarum GL64 and were placed inside coated-wire mesh cylinders (liveboxes) in the sham and disease challenge tanks, respectively. Mortalities in both tanks were recorded for 285 d. Any mortalities among the livebox fish were replaced with an appropriate cohort (infected with R. salmoninarum or healthy) fish. None of the bacterins evaluated in this study induced protective immunity against the R. salmoninarum shed from the infected livebox fish. The percentage survival within the test groups in the R. salmoninarum challenge tank ranged from 59% (heated Rs MT239 bacterin) to 81% (PBS emulsified with FIA). There were no differences in the percentage survival among the PBS-, PBS/FIA-, r-p57- and Renogen-injected groups. There also were no differences in survival among the bacterin groups, regardless of whether the bacterial cells had been heated or left untreated prior to injection. PMID- 15819431 TI - Production of monoclonal antibodies for detection of Vibrio harveyi. AB - Monoclonal antibodies (MAbs) against Vibrio harveyi were produced from mice immunized with heat-killed and SDS-mercaptoethanol-treated highly virulent V. harveyi 639. Fifteen MAbs were selected and sorted into 6 groups according to their specificity to various proteins of apparent molecular weight ranging from 8 to 49 kDa. Some antibodies were used for detection of V. harveyi at concentrations as low as 10(4) CFU ml(-1) using immunodot blots. Most of the selected MAbs did not show cross-reactivity to other Vibrio species and other gram-negative bacteria tested. Only 1 MAb (VH39-4E) showed slight cross reactivity to Aeromonas hydrophila. Another MAb (VH24-8H) bound lightly to V. harveyi 1526 but strongly to V. harveyi 639, allowing rapid differentiation. Two of the MAb groups were used to localize V. harveyi in tissues of infected black tiger shrimp Penaeus monodon by immunohistochemistry. This study demonstrates the versatility of a highly specific immunological tool for the detection of V. harveyi in aquaculture and opens the way for further development of convenient test kits. PMID- 15819432 TI - Effects of gill abrasion and experimental infection with Tenacibaculum maritimum on the respiratory physiology of Atlantic salmon Salmo salar affected by amoebic gill disease. AB - The effects of gill abrasion and experimental infection with Tenacibaculum maritimum were assessed in Atlantic salmon Salmo salar with underlying amoebic gill disease. The respiratory and acid-base parameters arterial oxygen tension (P(a)O2), arterial whole blood oxygen content (C(a)O2), arterial pH (pHa), haematocrit and haemoglobin concentrations were measured at intervals over a 48 h recovery period following surgical cannulation of the dorsal aorta. Mortality rates over the recovery period were variable, with gill abrasion and inoculation with T. maritimum causing the highest initial mortality rate and unabraded, uninoculated controls showing the lowest overall mortality rate. Fish with abraded gills tended to show reduced P(a)O2 and lower C(a)O2 compared with unabraded fish. Infection with T. maritimum had no effect on P(a)O2 or C(a)O2. All fish showed an initial alkalosis at 24 h post-surgery/inoculation which was more pronounced in fish inoculated with T. maritimum. There were no significant effects of gill abrasion or infection upon the ratio of oxygen specifically bound to haemoglobin or mean cellular haemoglobin concentration. Histologically, 48 h following surgery, abraded gills showed multifocal hyperplastic lesions with pronounced branchial congestion and telangiectasis, and those inoculated with T. maritimum exhibited focal areas of branchial necrosis and erosion associated with filamentous bacterial mats. All fish examined showed signs of amoebic gill disease with multifocal hyperplastic and spongious lesions with parasome containing amoeba associated with the gill epithelium. The results suggest that respiratory compromise occurred as a consequence of gill abrasion rather than infection with T. maritimum. PMID- 15819433 TI - Questionnaire-based risk assessment for amoebic gill disease (AGD) and evaluation of freshwater bathing efficacy of reared Atlantic salmon Salmo salar. AB - Factors causing amoebic gill disease (AGD), the main disease affecting the salmonid industry in Tasmania, are largely unknown. Managers of 57 sites, from 4 countries, in both the Southern and Northern Hemisphere were questioned regarding the 2000 smolt year-class Atlantic salmon. Questions included site characteristics, fish population characteristics, husbandry and management programs, through to freshwater bathing characteristics. Possible risk factors for AGD outbreaks that were identified in this study using farm managers' opinions were mixed sex stocks (Tasmania), downstream positioning from other salmonid farming sites (pooled), tidal sites (Tasmania), depth of site (pooled), rate of salinity change in halocline and depth of halocline (pooled, Tasmania), phytoplankton blooms (pooled, Tasmania), antifouling paints (Tasmania, overseas) and automated feeding (pooled, overseas). The information from farm managers suggested that freshwater bathing is less effective with smaller bathing cages and lower levels of saturated oxygen in the bathing water. Risk factors could not always be determined due to the complex interactions between the biological and chemical environment, host and pathogenic factors, coupled with low sample numbers. PMID- 15819434 TI - Anguillicola papernai (Nematoda: Anguillicolidae) and other helminths parasitizing the African longfin eel Anguilla mossambica. AB - The swim bladder nematode Anguillicola papernai Moravec & Taraschewski, 1988 has been investigated as regards its occurrence in longfin eels Anguilla mossambica (Peters) in rivers in South Africa. A. papernai revealed a prevalence of around 50% and a mean intensity of about 6 adult worms at 1 sampling site but were less abundant in 3 others. Field observations suggest a more narrow habitat preference than that of Anguillicola crassus and a seasonal pattern of abundance. African longfin eels harboured a poor helminth community. In addition to A. papernai, 2 gastro-intestinal nematodes occurred, the stomach worm Heliconema longissimum Ortlepp, 1923 as the dominant species, and the intestinal Paraquimperia africana Moravec, Boomker & Taraschewski, 2000. Experiments were undertaken using European eels Anguilla anguilla (Linnaeus) and copepods as laboratory hosts. The morphology of larvae and adult parasites obtained from these experimental hosts is described. The ultrastructure of adult worms recovered from wild longfin eels was studied. The 'papilla-like excrescences of fibrous structure' on the adult worms' cuticle, as mentioned in the original description, are in fact the attachment points of thick cords of fibers interconnecting the epicuticle with the hypodermis. Such a structure has not yet been described from any other species of Anguillicola Yamaguti, 1935. At present in South Africa, Mozambique and Madagascar attempts are on the way to establish an eel management like in Asia and Europe including eel farming. In this context, care should be taken to prevent the introduction of non-endemic eel parasites into Africa and Madagascar. On the other hand, the future commercial management of African eel species should not lead to the spread of A. papernai or other parasites of African eel species to Europe or elsewhere. In this study A. papernai has been experimentally demonstrated to be capable of reproducing in the European eel and of using European copepods as intermediate hosts. PMID- 15819435 TI - Parvicapsula pseudobranchicola (Myxosporea) in farmed Atlantic salmon Salmo salar. Tissue distribution, diagnosis and phylogeny. AB - Parvicapsula pseudobranchicola infections in farmed Atlantic salmon in Norway are associated with low-grade to significant mortalities. The parasite is found as mature spores in pseudobranchs, but has also been detected in the gills, liver and kidney. Diagnosis has relied on the detection of Parvicapsula spores, with the pseudobranch being the preferred organ. A better understanding of the epizootiology of this myxosporean is a prerequisite for appropriate management and control. Hence, early detection of infections and life cycle studies are needed. We sequenced the small subunit (ssu) rDNA (18S) from P. pseudobranchicola and developed a sensitive diagnostic PCR protocol. This allowed us to (1) identify appropriate tissues for diagnostic assays, (2) examine the intraspecific variation in ssu rDNA in the parasite's Norwegian range, (3) examine annelid potential primary hosts and (4) obtain additional ssu rDNA sequences of marine Parvicapsula species to perform a phylogenetic study. Primers were constructed targeting the ssu rDNA from P. minibicornis. With these we obtained a partial ssu sequence of the P. pseudobranchicola type isolate. A new set of primers (PCF3/PCR3) was constructed for diagnostic purposes. These were tested against DNA from the host and several myxozoan species infecting Norwegian salmon. The primers give a positive product of 203 bp and pick out P. pseudobranchicola in salmnonids. They also amplify the congeners P. unicornis and P. asymmetrica infecting unrelated fish. The PCR protocol developed showed a greater sensitivity than light microscopy. The pseudobranchs were always positive and are the recommended organ for PCR diagnostics. There was no sequence variation between geographic isolates from farmed salmon. Preliminary examinations of marine polychaetes and oligochaetes collected from farm sites with parvicapsulose problems were negative. A comparison of the sequence of the ssu rDNA from P. pseudobranchicola with that of other myxozoans shows that it groups closely together with P. unicornis and P. asymmetrica. The closest relative to this group is P. minibicornis. PMID- 15819436 TI - Revision of Myxobolus heterosporus Baker, 1963 (syn. Myxosoma Heterospora) (Myxozoa: Myxosporea) in African records. AB - There is uncertainty regarding the validity of Myxobolus heterosporus Baker, 1963. The present study revises the taxonomy, using specimens isolated from plasmodia situated in the infected cornea of Oreochromis aureus, O. niloticus or Tilapia zillii inhabiting the River Nile, Egypt. In addition, histological effects of the parasite on the infected tissue were examined. The spores of M. heterosporus had a variety of shapes expressing remarkable heteromorphism. Five main Myxobolus-like spore types and tailed-spores were found. All forms were photographed, measured, sketched and described. Light and electron microscopy supported that spores of a Myxobolus-like morphology co-existed with so-called tailed-spores in one plasmodium. Some transitional stages from Myxobolus-like spore types to tailed-spores were observed. Therefore, some tailed-spores may be simply heteromorphs of Myxobolus. PMID- 15819437 TI - Myxidium mackiei (Myxosporea) in Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii: parasite-host interaction and ultrastructure. AB - Myxosporeans are common parasites of fish, and uncommon parasites of amphibians, reptiles and invertebrates, that can cause significant morbidity and mortality. The common genus Myxidium infects the excretory system of turtles, yet knowledge of its pathogenicity in these hosts is limited. We offer new knowledge of morphological and ultrastructural aspects of host-parasite interactions in Myxidium infections from our recent diagnostic investigations on captive freshwater turtles listed in CITES (Appendix II). We investigated the cause of death of 2 adult Indo-Gangetic flap-shelled turtles Lissemys punctata andersonii from a zoo collection. After post-mortem examination, tissues were processed for histopathology, and special stains were used to demonstrate morphology of myxosporean spores. Additional kidney tissue, immersion-fixed in formalin, was processed for transmission electron microscopy. Both turtles were infected with a myxosporidian, Myxidium mackiei, in the kidney, which occluded 5 to 10% of the renal proximal convoluted tubules. The polysporic plasmodia contained pairs of developing and mature spores. Each mature, spindle-shaped spore had 2 asymmetric valves (1 overlapping, 1 overlapped), with 10 to 13 and 10 to 14 longitudinal ridges per valve, and 2 polar capsules each containing a polar filament with 4 to 5 turns. A pair of spores, each surrounded by a membrane-bound electron-lucent matrix, lay in an enclosing cell within the plasmodium. Regions of the parasite host interface consisted of undulations of the parasite surface, with intense pinocytotic activity beneath, intermingled with the hosts' microvilli, and endocytotic channels at the apex of renal epithelial cells. The microvilli of the renal epithelial cells of infected tubules were frequently sheared or compressed, or occasionally missing; we did not detect other pathology induced by the parasite. Our report of M. mackiei in L. punctata is a new host record. Both individuals also had disseminated pale yellow nodules (bacterial granulomas) present in lung, heart, kidney, and skeletal muscle, and both were infected with coccidia (tentatively identified as Eimeria sp.) in multiple organs. The cause of death for one turtle was septicemia, but remained unknown for the other individual. PMID- 15819438 TI - Occurrence of the haemocyte parasite Bonamia sp. in flat oysters Ostrea puelchana farmed in San Antonio Bay (Argentina). AB - Culture of native flat oysters Ostrea puelchana d'Orbigny in San Antonio Bay (San Matias Gulf, Argentina) began in 1995. After elevated mortality (33%) occurred in September 1996, 18 mo after immersion, histopathological analysis and evaluation of parasitic prevalence was carried out. In October 1997, after 31 mo of cultivation, cumulative mortality was 80%, and in December of the same year, when individuals reached marketable size, mortality was 95% and culture was discontinued. The present study describes the haemocytic parasitism that affected O. puelchana, and suggests that a Bonamia sp. was the etiological agent. This parasite should be considered as a different species from Bonamia sp. detected in Australia and New Zealand until more studies are made to determine the correct taxonomy. This work constitutes the first record of this haemocyte parasite in flat oysters from the Argentinean coast. PMID- 15819439 TI - Genetic characterization of the lobster pathogen Aerococcus viridans var. homari by 16S rRNA gene sequence and RAPD. AB - A combination of 16S rRNA sequencing and random amplified polymorphic DNA (RAPD) analysis was used to evaluate the genetic diversity within Aerococcus viridans var. homari, the causative agent of gaffkemia in lobsters. A collection of 7 A. viridans var. homari strains and 2 avirulent A. viridans-like cocci isolated from homarid lobsters harvested from different regions on the Atlantic Coast of North America were analyzed. The isolates are separated geographically and temporally between the years 1947 and 2000. Sequencing of 16S rRNA genes confirmed the inclusion of all 9 isolates in the monophyletic A. viridans clade (99.8 to 100% similarity). RAPD analysis revealed that the 9 A. viridans var. homari isolates could be separated into 2 distinct subtypes. Subtype 1 included the 7 pathogenic lobster isolates and constituted a homogeneous group regardless of their geographical, temporal or virulence differences. Subtype 2 contained the 2 avirulent A. viridans-like cocci that had distinct RAPD patterns and clustered separately with the non-marine A. viridans. RAPD analysis represented a useful method for determining molecular subtyping for the intraspecific classification and epidemiological investigations of A. viridans var. homari. PMID- 15819440 TI - Dermatitis, branchitis and mortality in empire gudgeon Hypseleotris compressa exposed naturally to runoff from acid sulfate soils. AB - Severe dermatitis and branchitis are described in a wild population of empire gudgeon Hypseleotris compressa, an Australian eleotrid, exposed naturally to runoff from acid sulfate soils (ASS) in a drained estuarine embayment in eastern Australia. After at least 2 d exposure to pH < 4, and up to 7 d exposure to pH < 6, approximately 50% of the fish sampled had moderate to severe diffuse epidermal hyperplasia, usually at scale margins, and scattered areas of moderate to severe, focal to locally extensive, subacute, necrotising dermatitis. Saprolegnia spp. had invaded epidermis in some inflamed areas. In gills, there was moderate to severe hyperplasia and necrosis of secondary lamellar epithelium, with fusion of adjacent secondary lamellae. Inorganic monomeric aluminium and calcium concentrations in water at the site during the event were 27.7 and 16.6 mg l(-1), respectively. Large numbers of empire gudgeons at the study site had died after at least 8 d exposure to pH < 4, and up to 13 d exposure to pH < 6. These findings provide clear evidence that acidification of estuarine systems by runoff from ASS has deleterious effects on aquatic biota. Furthermore, study findings suggest a mechanism whereby lesions of epizootic ulcerative syndrome (EUS) may be initiated in estuarine fishes by a combination of sublethal exposure to ASS runoff and Aphanomyces invadans infection, a suggestion consistent with the geographic and temporal distribution of EUS outbreaks in Australian estuaries. PMID- 15819441 TI - Detection of hepatopancreatic parvovirus (HPV) in wild shrimp from India by nested polymerase chain reaction (PCR). AB - The prevalence of hepatopancreatic parvovirus (HPV) in wild penaeid shrimp samples from India was studied by nested polymerase chain reaction (PCR) using primers designed in our laboratory. The virus could be detected in 9 out of 119 samples by non-nested PCR. However, by nested PCR 69 out of 119 samples were positive. The PCR results were confirmed by hybridization with digoxigenin labelled DNA probe. Shrimp species positive by non-nested PCR included Penaeus monodon, Penaeus indicus and Penaeus semisulcatus and by nested PCR Parapenaeopsis stylifera, Penaeus japonicus, Metapenaeus monoceros, M. affinis, M. elegans, M. dobsoni, M. ensis and Solenocera choprai. This is the first report on the prevalence of HPV in captured wild shrimp from India. PMID- 15819442 TI - In situ hybridization demonstrates that Litopenaeus vannamei, L. stylirostris and Penaeus monodon are susceptible to experimental infection with infectious myonecrosis virus (IMNV). AB - Infectious myonecrosis virus (IMNV) was recently found to be the cause of necrosis in the skeletal muscle of farm-reared Litopenaeus vannamei from northeastern Brazil. Nucleic acid extracted from semi-purified IMN virions showed that this virus contains a 7.5 kb RNA genome. A cDNA library was constructed, and a clone, designated as IMNV-317, was labeled with digoxigenin-11-dUTP and used as a gene probe for in situ hybridization (ISH). This probe specifically detected IMNV in infected tissues. To determine the susceptibility of 3 species of penaeid shrimp (L. vannamei, L. stylirostris, Penaeus monodon) to IMNV infection, juveniles were injected with purified virions and observed for clinical signs of infection and mortality over a 4 wk period. All L. vannamei exhibited typical lesions after 6 d, and lesions were visible in all L. stylirostris by Day 13. The clinical signs of opaque muscle were not seen in P. monodon, due to their highly pigmented exoskeleton precluding visual detection of lesions. Moderate mortality (20%) occurred in infected L. vannamei. No mortalities were observed in either L. stylirostris or P. monodon. Histological examination and ISH indicated that all 3 species are susceptible to IMNV infection. Using ISH, IMNV was detected in tissues including the skeletal muscle, lymphoid organ, hindgut, and phagocytic cells within the hepatopancreas and heart. In all 3 species, skeletal muscle cells produced the strongest ISH reactions. Based on the onset of clinical signs of infection and mortality, L. vannamei appears to be the most susceptible of these 3 species to IMNV infection. PMID- 15819443 TI - Epitopes associated with mature spores not recognized on Kudoa thyrsites from recently infected Atlantic salmon smolts. AB - Atlantic salmon Salmo salar skeletal muscle was examined for Kudoa thyrsites by polymerase chain reaction (PCR) and positive fish were further examined by in situ hybridization (ISH) and immunohistochemistry (IHC). The infection was detected in 42% of salmon by PCR following a 60 d exposure to infective seawater at a temperature of 10 degrees C (= 600 degree-days, degreeD). The parasite was detected by ISH in skeletal and cardiac muscle but not in gill, kidney, spleen, liver, stomach, intestine, pyloric caeca and skin. None of 4 monoclonal antibodies (2F4, 4H2, 1H2, 3E8) raised against mature K. thyrsites spores reacted with the stages identified by ISH following a 600 degreeD exposure, but they did react with ISH-identified stages following a 1600 degreeD exposure. In contrast, a polyclonal antibody reacted with K. thyrsites stages in salmon with both 600 and 1600 degreeD exposures, suggesting that the parasite observed in 600 degreeD infections represents an antigenically distinct developmental stage of K. thyrsites. PMID- 15819444 TI - Finding the right medicine: skillful means. PMID- 15819445 TI - Alternative therapies--a moving target. PMID- 15819446 TI - Naturopathic medicine--a 10-year perspective (from a 35-year view). AB - The decade of Alternative Therapies in Health and Medicine's existence has seen dramatic advancement of the naturopathic profession in all areas: research, quality of practice, quality of education, licensure, inclusion in the political process, and public acceptance. Naturopathic doctors are now valued in the marketplace and in policy as a legitimate part of an overall system of healthcare. Momentum, infrastructure, acceptance, and conceptualization are all foundational to the future. Alternative Therapies in Health and Medicine was one of the first journals to invite naturopathic doctors onto its Editorial Board (i.e., Pizzorno, Standish). Our profession thanks and applauds you for inviting us. PMID- 15819447 TI - Homeopathy: increasing scientific validation. PMID- 15819448 TI - Meditation can reduce habitual responding. AB - CONTEXT: Although cognitive aspects of meditation underlie much of its clinical application, very little research has examined meditation's cognitive consequences. This investigation provides experimental support for the idea that meditation leads to a reduction in habitual responding using randomly selected subjects, a secular form of meditation, and a full experimental design. OBJECTIVE: To test the hypothesis that meditation leads to a reduction in habitual responding. DESIGN: Studies 1 and 2 each incorporated pre-test and post test designs with a 20-minute intervening attention task (meditation, rest, or a cognitive control). SETTING: Yale University in New Haven, Conn, and the University of California, Berkeley. PARTICIPANTS: One hundred and twenty and 90 undergraduates participated in Studies 1 and 2, respectively. MAIN OUTCOME MEASURES: Stroop and Word Production (category generation and stem-completion) tasks assessed habitual responding in Study 1. Galvanic Skin response measured arousal in Study 1. The category generation task assessed habitual responding in Study 2. Tellegen's Absorption Scale (TAS) measured attention ability. RESULTS: In Study 1, meditation participants showed a reduction in habitual responding on the Stroop task as compared to controls. Study 1 revealed no statistically significant effects in the word production task. Stroop task performance was not mediated by arousal reduction. In Study 2, meditation participants showed a reduction in habitual responding on the category production task. Specifically, when participants generated either typical or atypical items, on average, meditation participants produced more atypical items than controls. Category production performance was not mediated by Tellegen's Absorption Scale (TAS) scores. Overall, high TAS scores were related to atypical responding. CONCLUSION: Across cognitive tasks, when participants understood that the goal was to respond non-habitually, meditation reduced habitual responding. PMID- 15819449 TI - Tai Chi for vestibulopathic balance dysfunction: a case study. PMID- 15819450 TI - Jeremy Geffen, MD: advancing the concept of multi-dimensional medicine. Interview by Karolyn A. Gazella. PMID- 15819451 TI - Bone lesions in multiple myeloma--the OPG/RANK-ligand system. AB - Multiple myeloma has recently been found to induce considerable imbalance in the newly identified system of osteoprotegerin (OPG), receptor activator of nuclear factor KB ligand (RANKL) and RANK. The binding of RANKL to RANK on the surface of osteoclastic precursors in the presence of m-CSF activates the signalling pathways for differentiation and proliferation of an osteoclastic line. OPG is a decoy circulating receptor for RANKL which blocks its binding to RANK. There are at least three mechanisms by which myeloma cells affects the OPG/ RANKL/RANK system: 1: The adhesion between the myeloma / stromal cells and the osteoblastic precursors stimulates the system by increasing the production of RANKL. 2: Some myeloma lines produce independently membrane-bound or free RANKL. 3: The normal and mutated plasma cells bind, degrade and block the OPG production from the stromal cells. The OPG/RANKL/RANK system is the latest therapeutic target in the treatment of myeloma bone disease. The first results from the application of a synthetic analogue of OPG, as well as of RANKL antagonists or RANK inhibitors show decrease of the number of osteoclasts, osteolytic lesions and M-gradient. PMID- 15819452 TI - Long-term changes in dyspnea, lung function, and exercise capacity in COPD patients. AB - OBJECTIVE: To examine longitudinal changes in dyspnea, lung function, and exercise capacity in COPD patients and to compare baseline data of frequent and infrequent exacerbators at trial entry. METHODS: Nineteen stable COPD patients without significant co-morbidity (age, 58.4 +/- 8.4 years; FEV1% = 33 +/- 12%; mean +/- SD) participated in the study. After a mean period of 36 months (range = 24 - 49) the patients were retested using an identical protocol. RESULTS: Repeated measures analysis showed that there was significant deterioration of FEV1 L (from 1.028 +/- 0.349 to 0.928 +/- 0.307; p = 0.007), PImax cm H2O (from 61.9 +/- 24.2 to 42.0 +/- 22.1; p = 0.007), PaO2 mm Hg (from 69.0 +/- 8.6 to 60.1 +/- 6.8; p = 0.003), PaCO2 mm Hg (from 43.1 +/- 4.9 to 47.3 +/- 4.5; p = 0.001), ATS (from 2.4 +/- 1.0 to 2.8 +/- 0.8; p = 0.031), and 6MWD m (from 389 +/- 130 to 341 +/- 135; p = 0.014). There were also changes in IC, T(L,CO)/V(A), PAP and Borg, but they were not statistically significant. Differentiation of patients by frequency of exacerbations per year of observation (> 2 < or =) discriminated them with respect to functional parameters (FEV1, FVC, IC), dyspneic indices (ATS, VAS and Borg) and exercise capacity (6MWD) at the time of enrollment. CONCLUSIONS: 1) Lung function parameters, blood-gas and dyspneic indices, and exercise capacity decline over a mean period of 36 month in patients with COPD; 2) Patients with frequent exacerbations experience more dyspnea and have lower levels of lung function and exercise capacity at trial entry. PMID- 15819453 TI - Polymorphism of the serum alpha-amylase in the population of the South Central and South Eastern regions of Bulgaria and its relationship with the erythrocyte blood group systems ABO and MNSs. AB - INTRODUCTION: The goal of the study was to investigate the relationship between the polymorphism of alpha-amylase and the erythrocyte systems--ABO, MNSs, and determine their population distribution and individual phenotype and gene incidence. MATERIAL AND METHODS: Parallel examinations of identical blood samples were carried out to determine the phenotypes of alpha-amylase and ABO and MNSs blood group systems. Blood was obtained by venipuncture in the Serological Laboratory of the Department of Forensic Medicine at the Medical University of Plovdiv. Amylase phenotypes were determined by the authors' own modification of Kompf's method using continuous horizontal electrophoresis of blood serum on agarose gel. Only the classical phenotypes of the examined blood group systems- alpha-amylase, ABO and MNSs were determined in the study. RESULTS. The analysis of the data by chi-square analysis using Pearson criterion found no relationship between the polymorphism of the serum alpha-amylase and ABO and MNSs erythrocyte systems. The allele frequencies determined were 1(a) = 0.9595, 1(b) = 0.0388, 1(o) = 0.0017 for Amyl and 2(a) = 0.9579 and 2(b) = 0.0421 for Amy2. CONCLUSION: The difference between the observed and expected phenotypes of the examined blood group systems alpha-amylase, ABO, MNSs is short of statistical significance. This indicates genetic stability of the examined blood group systems in the studied population. PMID- 15819454 TI - An epidemiologic study of some characteristics of lung cancer. AB - OBJECTIVE: The principal aim of this study was to determine lung cancer distribution according to some epidemiologic and demographic variables in patients diagnosed to have this disease. METHODS: The study included 1731 patients diagnosed to have lung cancer between January 1, 1990 and December 31, 1999. Data were gathered from the patients' history of disease and from the bronchoscopy Record book. Establishment of significance in analyzed differences between some statistical series was done using Pearson's chi2 test and Kolmogoroff-Smirnoff test (K-S test) for one and two samples. RESULTS: The majority of male patients (out of a total of 1523 males) were in the age range of 60 to 64 years (21.7%). The age range of 65 to 69 was predominant in the female patients (16.3%). The distribution by sex and age showed significant differences (K-S; p < 0.05). 81.4% of the male patients and 39% of the female patients were smokers. This characteristic was significantly associated with the patients' sex (chi2 = 258.12; DF = 1; p < 0.01). Bronchoscopic localization of the pathological process was not significantly correlated with the habit of cigarette smoking (Kruskal-Wallis chi2 = 1.36; DF = 2; p > 0.05). Pathohistological diagnosis and smoking were significantly correlated (chi2 = 49.37; DF = 2; p < 0.01). CONCLUSIONS: In the study period, the men fell ill seven times as frequently as the women. The pathological process was most commonly localized in the right and left upper lobes. The predominant histologic type in men was squamous cell carcinoma (SQC), and in women--adenocarcinoma (ADC). Smokers were the greatest number in the patients with SQC and small cell carcinoma. ADC was most common in nonsmokers. PMID- 15819455 TI - Tidal breathing analysis in school-age children. Comparison with the parameters of forced expiration. AB - Tidal breathing analysis is a method which has the potential to be used for distinguishing and follow-up of airflow obstruction (AFO) in infants, children and critically ill patients. The aim of the present study was to analyse the tidal breathing parameters (TBP) in healthy and in asthmatic school-age children and to compare them with the parameters of forced expiration. SUBJECTS: Two hundred and twenty five healthy children and 100 asthmatics (7 to 14 years- old) took part in the present study. RESULTS: The results show that TBPs exhibit great inter- and intraindividual variability, even if the mean value of 10 consecutive breathing cycles is used. Parameters that reflect the tidal expiratory flow pattern--V(PTEF)/V(E) and T(PTEF)/T(E) demonstrate high variability and no correlation with age, sex and anthropometric parameters in healthy children. These indices are useful for detection of acute changes in bronchomotor tonus in asthmatics - V(PTEF)/V(E) = 36.1 +/- 6.6% vs. 32.6 +/- 6.2% (methacholine) vs. 37.4 +/- 7.5% (salbutamol) and T(PTEF)/T(E) = 34.2 +/- 6.2% vs. 28.6 +/- 7.8% vs. 35.3 +/- 7.5%, resp. (P < 0.05 everywhere; n = 34) as well as for discriminating a group of subjects with AFO vs. controls (V(PTEF)/V(E) = 30.9 +/- 6.5% vs. 35.3 +/- 8.0%; P = 0.005, and T(PTEF)/T(E) = 29.0 +/- 6.7% vs. 32.8 +/- 7.6%; P = 0.016). The evaluation of the area under the ROC curves (AUC) in the asthmatic group showed weak discriminative capacity of T(PTEF)/T(E) and V(PTEF)/V(E) in comparison to FEV1 (AUC of T(PTEF)/T(E) = 0.62; 95%CI 0.51-0.74). CONCLUSIONS: Tidal breathing parameters could add insight t.o the functional profile but are not capable of substituting forced expiration regarding detection of overt airflow obstruction in school-age children. PMID- 15819456 TI - Diagnostic value of the muscle fiber conduction velocity for evaluation of muscle hypotrophy. AB - AIM: To find an objective method of evaluation of the thigh muscle hypothrophy in gonarthrosis. MATERIAL AND METHODS: The subjects of the study were 24 patients with gonarthrosis (mean age 58.4 +/- 2.6 years). Of these 8 (33%) were males and 16 (67%) females. The following methods of examination were used: computed tomography (CT) of the thigh musculature--linear measurement and area of the cross-section of the vastus medialis muscle; electromyography--muscle fiber conduction velocity; and measurement of thigh girth. RESULTS: Significant reduction of the cross-section area and linear measurement of the vastus medialis muscle and significant delay of the muscle fiber conduction velocity was found in the symptomatic side compared with the asymptomatic side. No correlation was found between the tape measure of the thigh and cross-section area of the muscle. A significant correlation was found between the cross-section area of the muscle and the fast fiber conduction velocity as well as between the linear measurement of the vastus medialis muscle cross-section and fast fiber conduction velocity. CONCLUSION: The correlation between the macromorphological changes of the vastus medialis muscle assessed by computed tomography and the functional changes detected by electromyography allows the conclusion that the muscle fiber conduction velocity can be used as an objective method of evaluation of muscle atrophy in degenerative joint diseases. PMID- 15819457 TI - Fluconazole (Fungolon) in the treatment of onychomycoses. AB - Onychomycoses are fungal infections of the hand and foot nails, caused by dermatophytes, yeasts and molds. These conditions are difficult to treat, spontaneous remissions are rare and recurrences are not infrequent. AIM: The present study aimed at establishing the efficacy of pulse fluconazole therapy (200 mg once weekly for 4 months) in cases of hand nails infections and 200 mg once weekly for 6 months in cases of feet nail infections. MATERIAL AND METHODS: The study comprised 36 patients--23 with hand onychomycosis and 13 with feet onychomycosis (patients' age ranging from 22 to 67 years). More than 25% of the nail bed was affected in all cases. Diagnosis was based on direct microscopy of KOH treated specimens and by culturing on Sabouraud medium. RESULTS: Treatment outcome was categorized as follows: clinical cure--in 91.3% of the cases with hand onychomycosis and improvement--in 8.7% of the cases; eradication of fungal infection--in 95.6% of the cases. Feet onychomycosis--clinical cure--61.5% of the patients, improvement--in 38.5% and eradication of fungal infection--in 69.2 % of the cases. Fluconazole (Fungolon) has only been recently used in the treatment of nail fungal infections. So far, no standard dosage has been established. CONCLUSIONS: On the basis of the accumulated evidence it is concluded that pulse fluconazole (Fungolon) therapy--200 mg once weekly is effective and safe. However, for better disease outcome, especially in cases of foot nail infections- dosage of 300 mg once weekly and 300-450 mg once weekly (if molds are the offending agents) is recommended. PMID- 15819458 TI - Low back pain in physical education teachers. AB - The purpose of this study was to investigate the factors related to occupational low back pain (LBP) occurring in male physical education teachers (PET). The location was set in Athens and we used questionnaires to collect information. Analysis revealed that the prevalence of LBP during the survey was 63%. The correlation of personal factors and LBP showed that the prevalence of LBP was significantly higher in those PET (odds ratio, 2.5) who answered yes to the item "no personal training" than in the PET who did not. Moreover, many occupational factors are related to LBP. These were the items "lifting gym instruments" (odds ratio, 2.6) and "helping students into flexing posture" (odds ratio, 3.0). From the working conditions it was confirmed that the PET (odds ratio, 2.5) who "spent more than 35 hours per week in teaching physical education" were correlated with LBP. Our results suggest that PET, under the current working condition, are vulnerable to low back pain. An improvement of these conditions must be taken into account in order to prevent LBP. PMID- 15819459 TI - Sport related plantar fasciitis. Current diagnostic and therapeutic advances. AB - Plantar fasciitis is the most common diagnosis for pain in the inferior aspect of the heel among runners, accounting for 10% of injuries that occur in connection with running. The etiology of pain is multifactorial. The aim of the present study is to report our experience in the treatment of plantar fasciitis in athletes and compare our diagnostic strategy and treatment modalities with the current practice. MATERIAL AND METHODS: We treated 32 athletes with plantar fasciitis from 1997-2002. The diagnostic procedure included detailed history, clinical examination and imaging techniques. Conservative treatment consisted of anti-inflammatory drugs, stretching exercises, suitable training routines and special orthotic insoles, which offer a good chance of complete resolution of symptoms. Surgical fasciotomy should be reserved for use in patients in whom conservative measures have failed. RESULTS: Conservative treatment consisting of rest, anti-inflammatory medications, stretching exercises and special orthotic insoles was efficient in 26 patients (81%), while only 6 (19%) had to be treated surgically. CONCLUSION: Plantar fasciitis is a common disorder among athletes. Patient's history, clinical examination and common imaging techniques help to make the correct diagnosis. The patient should be treated conservatively at first and only in severe cases surgically. PMID- 15819460 TI - A reproducible model of cardiac hypertrophy in rats. Problems and obstacles we faced. AB - AIM: To develop a reproducible model of significant left ventricular hypertrophy in order to study the role played by the tumor suppressor protein 53 (P53) in the mechanisms of cardiac hypertrophy, the cross-talk with the other factors and the connection between expression and activity of P53, cardiac myocyte apoptosis and heart hypertrophy; to discuss the problems and obstacles we faced. METHODS: Male Wistar rats were used in the experiments. Left ventricular hypertrophy was induced by banding the abdominal aorta. Three series of the experiment were performed differing in the place of banding and the size of constriction in order to find the most suitable model for our further research. RESULTS: Forty five days after banding the abdominal aorta just below the diaphragm the cardiac index increased by 30% in comparison with that in the controls. CONCLUSION: We have developed a reproducible model of significant left ventricular hypertrophy in rats. The main advantages of the model are: a) it is technically simple; b) it may be realised in any laboratory all over the world; c) it allows the most common lab animals (male Wistar rats) to be used for studies without having to be subjected to thoracotomy and hence requiring a long recovery period. PMID- 15819461 TI - Absolute bioavailability of quinine formulations in Nigeria. AB - This study compared the absolute bioavailability of quinine sulphate as capsule and as tablet against the intravenous (i.v.) infusion of the drug in twelve male volunteers. Six of the volunteers received intravenous infusion over 4 h as well as the capsule formulation of the drug in a cross-over manner, while the other six received the tablet formulation. Blood samples were taken at predetermined time intervals and plasma analysed for quinine (QN) using reversed-phase HPLC method. QN was rapidly absorbed after the two oral formulations with average t(max) of 2.67 h for both capsule and tablet. The mean elimination half-life of QN from the i.v. and oral dosage forms varied between 10 and 13.5 hr and were not statistically different (P > 0.05). On the contrary, the maximum plasma concentration (C(max)) and area under the curve (AUC) from capsule were comparable to those from i.v. (P > 0.05), while these values were markedly higher than values from tablet formulation (P < 0.05). The therapeutic QN plasma levels were not achieved with the tablet formulation. The absolute bioavailability (F) were 73% (C.l., 53.3 - 92.4%) and 39 % (C.I., 21.7 - 56.6%) for the capsule and tablet respectively and the difference was significant (P < 0.05). The subtherapeutic levels obtained from the tablet form used in this study may cause treatment failure during malaria and caution should be taken when predictions are made from results obtained from different formulations of QN. PMID- 15819462 TI - Tooth size symmetry--a comparative analysis of tooth sizes among secondary school children. AB - A study was carried out to determine the mesio-distal crown dimensions of permanent teeth, and to find out if there were any symmetry of the tooth sizes in a population of Nigerian children. The sample consisted of 250 secondary school children (125 males and 125 females) randomly selected from 3 secondary schools within Ibadan metropolis. Their age ranged from 12-15 years the mean age was 12 +/- 0.5 years. The results showed that there were no significant differences (p > 0.05) between the right and left sides of the dental arch with the exception of the maxillary second premolars and the female mandibular canines. PMID- 15819463 TI - Determinants of survival in Nigerians with Burkitt's lymphoma. AB - There is paucity of information on the factors influencing survival of patients with Burkitt's lymphoma (BL) in Nigeria. This work was undertaken to examine the roles of presenting clinical and laboratory features in the survival of a number of Nigerian patients with BL. Confirmed cases of BL diagnosed in the hospital between January 1986 and December 1999 were studied. Diagnosis of the tumour was based on fine needle aspiration cytology of easily accessible mass in a large majority of patients. Median survival was calculated for all the dead patients using Kaplan-Meier technique. SPSS 10 statistical software and EPI 6.04b were used for all statistical calculations. A total of 213 patients were recruited over the 13-year period; with ages ranging from 3-45 years (median = 9) at diagnosis. The male-female ratio was 1.8:1. At the time of analysis in December 2001, 166 (77.9%) of patients had defaulted, 44 were known to have died and 3 were still alive. Only 81 (38%) of the patients had adequate chemotherapy (4-6 cycles). Fifty one (23.9%) patients had an initial complete remission. The mean survival of the 44 patients that died was 10.5 weeks (95% Cl = 3.9-17.1). It is concluded that survival in Burkitt's lymphoma was significantly better in patients who had adequate therapy, mean 40.8 +/- 12.2 (SE) weeks compared to those who had less than adequate cycles of chemotherapy, mean 3.7 +/- 1.0 (SE) weeks (p = 0.0000). Inadequate therapy and high default rate were partly responsible for the poor survival results obtained in this study. PMID- 15819464 TI - Nigerian back school model: development and effect on industrial workers' knowledge of back pain and back care. AB - Back schools are health education programmes on back pain. Many back schools have been developed for different populations since 1969 when the first one was developed in Sweden, but none for the Nigerian setting. The need to develop a back school that would be appropriate for the Nigerian environment was therefore identified. As a step towards its development, this preliminary study was carried out to determine the effect of a back school model (BSM) on some Nigerian industrial workers' knowledge of low back pain and back care. The subjects were 110 workers of a soap industry in Lagos, Nigeria. Apre-test, post-test quasi experimental design was used. The BSM consisted of two 45 minute teaching sessions on structure and functions of the back, epidemiology and causes of back pain, correct postures and demonstration of exercises that may prevent/alleviate back pain. Data on demographic information, low back pain (LBP) experience, knowledge of back structure and back care were collected using a questionnaire with closed ended questions, which was completed before, immediately after and 8 weeks after BSM administration. The results showed that the subjects' mean knowledge score increased significantly from an initial value of 16.1 + 5.3 to 24.0 + 5.6 (p < 0.05) immediately after and 23.1 + 3.9 (p < 0.05) 8 weeks after BSM administration. Educational attainment had no influence on subjects' knowledge scores before or 8 weeks after BSM administration. Reported experience of LBP and duration on the job had no significant influence on subjects' knowledge scores before, immediately or 8-weeks following BSM administration. It was concluded that the BSM was effective in improving the workers' knowledge of LBP and back care. We believe that this findingjustifies the effort to develop the Nigerian model of back school. PMID- 15819465 TI - Sexual networking among married men with wives of child bearing age in Ibadan City, Nigeria: report of a pilot study. AB - Following the Beijing Conference, it is desirable to empower men to play a more active and responsive role in promoting the health of family members and preventing disease. This cross sectional, community-based, ex-post factor, pilot study was designed to find out if traditional norms affect marital sexuality and also to identify sociodemographic factors associated with sexual networking among men. A total of 416 married men whose wives had delivered a baby in the last 36 months prior to the study were interviewed from randomly selected clusters in Ibadan, Nigeria. Of this number, the majority 336 (80.8%) had sex with pregnant wife in the last pregnancy and proportion of married men who had sex in the pregnancy with wives reduced modestly with increasing age of the men. With regards to sexual networking in pregnancy, 207 (49.8%) men reported having sex with someone else when wife was pregnant. Of this number 95 (45.9%) had it with steady girl friends, 56 (27.0%) with new girl friends, 50 (24.2%) with another wife and 6 (2.9%) with commercial sex workers. The prevalence for having sex with someone else in this period was lower in men from the higher socio-economic class (HSEC) when compared with the lower socio-economic class (LSEC)(chi2 = -9.89, P < 0.001). The middle socio-economic class also had a lower rate than the lower socio-economic class (chi2 = 6.28, P < 0.01). In addition, men with post secondary/University education had significantly lower rates for networking when their wives were pregnant compared with men of lower educational attainment (P < 0.05). Three hundred and eleven men (74.8%) reported that they observed some period ofpostpartum abstinence (PPA) with recently delivered wife, which ranged from 5 days to 72 months (Median was 7.5 months). The highest PPA rates were seen in men with no formal education, those from lower SEC and in men who embraced traditional religions. Issues that have to be addressed in more detail in the follow up study include understanding why men network as traditional proscriptions are generally not adhered to. Determining the relationship between length of postpartum abstinence and sexual networking and implication of this behaviour in the spread of sexually transmitted diseases and fertility control if postpartum abstinence period is significantly reduced or increased. In this regard, qualitative research as well as quantitative research should be carried out so that the entire study is not left within the realm of a quantitative study, which may be inadequate for explaining social and demographic data. PMID- 15819466 TI - Socio-demographic determinants of anaemia in pregnancy at primary care level: a study in urban and rural Oyo State, Nigeria. AB - Anaemia in pregnancy is a serious condition contributing to maternal mortality, morbidity and fetal morbidity. Data however available on this condition in Nigeria is mainly hospital based and biased towards women with high-risk pregnancies. This study was carried out to determine the prevalence of anaemia and identify sociodemographic factors contributing to anaemia among low risk pregnant women attending primary care facilities. Five hundred and ninety seven pregnant women attending randomly selected primary care centres in urban and rural areas in Oyo State, Nigeria were enrolled and followed up till delivery of their babies. One hundred and ninety six (32.8%) of the mothers were anaemic. Prevalence of anaemia decreased with increasing maternal age in both urban and rural areas. Regression analysis showed that urban mothers (P = 0.003) and those who booked late in pregnancy (P = 0.048) were significantly more likely to be anaemic. Mothers with birth intervals 24 -35 months and women between the ages 20 29 years (P = 0.011) had a lower risk for anaemia. Prevalence of anaemia in pregnancy is still high in Nigeria. Effort should be made to ensure that mothers attend antenatal clinic early. Longer birth spacing should be encouraged through the use of effective contraception. PMID- 15819467 TI - Preliminary studies on the prevalence and distribution of urinary schistosomiasis in Ondo State, Nigeria. AB - The prevalence of S. haematobium infection in 18 Local Government Area (LGA) in Ondo state was determined by examining pupils from randomly selected schools in the state. A total of 4266 urine samples were examined and 1303 (30.5%) were infected with the parasite. The infection was recorded in both rural and urban communities in each LGA. The overall prevalence was significantly higher in males (33.7%) than in females (26.0%) (P = -0.001). The LGA exhibiting the highest overall prevalence of infection was Akure north (86.2%; Cl, 0.82-0.9). The infection was most common in subjects between the ages of 11 and 15 years. Seven species of aquatic snails were encountered and B. (P). globosus was the most abundant and widespread species. The Owena river/dam in Idanre LGA was richest in molluscan fauna. B. (P). globosus and Biomphalaria pfeifferi shedding cercariae were recovered from Owena river and dam. The study shows that there were spatial heterogeneities in the prevalence of infections in both human population and the snail intermediate hosts. The implications of the clustering nature of the disease, for a successful control program in the state, were discussed. PMID- 15819468 TI - Nasal foreign bodies in the African children. AB - Nasal foreign bodies are very common in daily clinical practice. Their simplicity in pathology and diagnosis often gives the wrong impression of little or no risk of complications. A 5-year (1998 - 2002) prospective study of 106 patients with nasal foreign bodies was done to evaluate and present the patterns, possible challenges and complications or problems in the management of this condition in the Nigerian Africans. The male to female ratio was 1:1.26 (M 47; F 59) and with an average age of 3 years. The duration of symptoms ranged from degrees hour to 4 years with 74 (69.8%) presenting within 24 hours and 27 (25.5%) presenting after 24 hours. The most common nasal foreign bodies were seeds 34 (32.1%), polyurethane foams 12 (11.3%), stones 11 (10.4%), plastic 10 (9.4%), beads 6 (5.7%) and erasers 6 (5.7%). The objects were found in the right nasal cavity in 63 (59.4%) cases while 43 (40.6%) in the left nasal cavity. The various clinical presentations were history of insertion of foreign bodies 91 (85.8%), 15(14.2%) with no history of insertion, mucopurulent nasal discharge 25 (23.6%), foul nasal odour 10 (9.4%), epistaxis 6 (5.7%), nasal obstruction and mouth breathing 3 (2.8%) and 2 (1.9%) cases respectively. The main complications were nasal infections (23.6%), epistaxis (5.7%), and purulent maxillary sinusitis (1.9%) seen in this study. These are preventable complications if the patients present early to the hospital. The absence of enough E.N.T. specialists however still plagues developing countries like Nigeria. A call is therefore made for more specialists in this area for early detections and care of these cases. PMID- 15819469 TI - Risk factors for HIV infection among special treatment clinic attendees in Ibadan, Nigeria. AB - Control of sexually transmitted infections (STIs) is an important factor in the prevention of sexual transmission of HIV infection. Despite a close link between the two disease entities, not enough data are however available on risk factors for HIV infection among STI clinic patients in Nigeria. This information is important for planning preventive measures. All consenting patients that attended special treatment clinic (STC), University College Hospital (UCH), Ibadan from March to November 2001 were enrolled into the study. Using a structured interview assisted questionnaire, information on socio-demographic characteristics, sexual and reproductive history and other potential risk factors for HIV infection were obtained. Urethral swab, endocervical swab and high vaginal swab were also obtained and processed by standard laboratory methods. HIV tests were carried out by double ELISA tests. Of the 210 subjects, 180 (85.7%) were diagnosed as having STI while 46 (21.9%) screened positive for HIV. Co-infection was found in 41(19.5%) of all subjects. The risk of HIV was higher in subjects with STIs. Multivariate logistic regression analysis indicated that subjects who were not infected with gonorrhoea (OR = 0.685, 95% CI = 0.47- 0.99, p = 0.045) and those with negative history of blood transfusion (OR= 0.1,95% CI = 0.02-0.5, p=0.009) were less at risk of HIV infection while those with lower occupation had a higher risk (OR = 1.126,95% CI = 0.32-3.9, p = 0.085). The results of this study emphasize the need for adequate management of STIs and the importance of upgrading blood transfusion services in order to curtail the spread of HIV infection. PMID- 15819470 TI - Femoral head diameter in subcapital fracture of the femur in Ibadan, Nigeria. AB - Subcapital fracture of the femur is common in the elderly patients though the incidence is less in our environment than in the western world. Primary prosthetic replacement is the method of treatment in the majority of the patients. To facilitate this, a foreknowledge of the femoral head diameter is necessary. To determine the diameter of the femoral head and its magnification on radiograph among patients with hip fractures seen in South-West Nigeria, all patients with subcapital fracture of the neck of femur seen between March 1997 and February 2002 were included in the study. The femoral heads were measured on the radiographic film using a transparent ruler and after extraction of the femoral head during surgery using callipers. There were 25 patients in all, the mean age of patients was 73.2 years. Fall at home accounted for 70% of the cases. The femoral head diameter was between 42 mm and 50 mm in 92% of patients while magnification of femoral head diameter on radiograph was 10-14% in 92% of cases. The usaof 10-14% magnification of femoral head on radiograph would help the surgeon determine the size of prosthetic head before commencement of surgery. PMID- 15819471 TI - Utilization of obstetric care services in a rural community in southwestern Nigeria. AB - Utilization of Obstetric Services in Nigeria is very low with only a third of the deliveries being conducted under supervision of trained health personnel. Consequently maternal and infant mortality rates are unacceptably high at 1000/100,000 and 100/1000 live births per year respectively. We conducted a cross sectional survey of 100 randomly sampled women in a rural community in Oyo State in Nigeria to study the pattern of utilization of antenatal, delivery and postnatal care services in the community. Results showed that utilization of antenatal care services to be relatively high but most of the respondents delivered at home without the supervision of trained health personnel. This poor utilization of delivery services was attributed to advanced labour and perceived poor quality of the health facilities in the community. Although postnatal care was given to the respondents, it did not include advice on family planning/child spacing. The variables found to have statistically significant association with seeking antenatal care were age and educational attainment (P < 0.0005). Educational attainment also significantly affects the respondents' choice of the place of delivery (P < 0.005). We recommend operations research to assess and improve the quality of existing health facilities and training/retraining of antenatal care providers on interpersonal communication skills, early recognition of labour and seeking delivery care. This training should also include providing advice on child spacing and use of obstetrics services provided. PMID- 15819472 TI - Alterations in the expression of androgen receptor, wild type-epidermal growth factor receptor and a mutant epidermal growth factor receptor in human prostate cancer. AB - Prostatic carcinogenesis has been associated with alterations in the expression of the androgen receptor (AR) and the epidermal growth factor receptor (WT-EGFR), and over-expression of the constitutively active variant epidermal growth factor receptor (EGFRvIII). Changes in the expression of AR, WT-EGFR and EGFRvIII were evaluated in serial sections from 26 normal and 26 benign hyperplastic and 50 prostate cancer tissues using specific immunostaining techniques. The loss of AR expression in peri-epithelial stroma as prostatic tissues de-differentiated correlated strongly with the depletion of WT-EGFR and with increasing expression of the EGFRvIII in the adjacent epithelium. In contrast, changes in epithelial AR immunopositivity in these tissues correlated weakly with the changes in normal and variant EGFR levels. This is the first report correlating the changes in the expression of these three proteins in archival material from the different human prostatic tissue histotypes. The loss of expression of proteins that contribute to the regulation of prostatic homeostasis (AR and WT-EGFR) correlates strongly with the expression of a constitutively active variant EGF receptor (EGFRvIII) in human prostate cancer. These changes occur at an early stage of neoplastic transformation and may contribute to the progression of the disease to hormone independence. PMID- 15819473 TI - Fine needle aspiration cytology diagnosis of liver diseases in the University of Maiduguri Teaching Hospital, Maiduguri. AB - The study is aimed at classifying liver diseases diagnosed by fine needle aspiration cytology (FNAC) in our environment and to emphasize the importance of the procedure for management of our patients. Forty-two patients were referred by the physicians for fine needle aspiration cytology diagnosis in Histopathology Department, University of Maiduguri Teaching Hospital between January 2001 and December 2003. There were 30 males and 12 females (2.5:1) with mean age of 44.7 years. The peak age is in the 5th decade of life. The commonest hepatic disease is primary hepatic malignancy, which accounted for 78.6% (33 patients); 3 suspicious of malignancy, one case each of metastatic carcinoma, amoebic liver abscess; liver storage disease and large cell dysplasia. The technique has gained popularity in our center because it is cheap, convenient, minimally invasive, quick and has high precision in obtaining samples. We therefore recommend this procedure in centers where liver diseases are common as a means of initial investigation. PMID- 15819474 TI - Evaluation of modified short course chemotherapy in active pulmonary tuberculosis patients with human immunodeficiency virus infection in University College Hospital, Ibadan, Nigeria--a preliminary report. AB - Over the period, 1st October 1999 to 30th April 2002 a clinical trial of the modified short-course chemotherapy (SCC) in newly diagnosed cases of pulmonary tuberculosis with human immunodeficiency virus (HIV) infection in Ibadan, Nigeria was carried out. The modified SCC used was adopted by World Health Organisation (WHO)/International Union against Tuberculosis and Lung Diseases (IUALTD) for developing countries and also by the Nigerian National Tuberculosis and Leprosy Control Programmed (NTLCP). The regimen used consisted of ethambutol (E), isoniazid (H), rifampicin (R) and pyrazinamide (Z) in the intensive phase of 2 months. The continuation phase was 6 months of ethambutol (E) and isoniazid(H), i.e. 2EHRZ/6EH. Sputum conversion was 90% at the second month of treatment and there was no bacteriological relapse after 18 months of follow-up. Side effects were few and consisted mainly of acne vulgaris in 20 (22.5%) of 89 patients during the continuation phase. It is concluded that this modified 8-month chemotherapy regimen adopted by NTLCP is efficacious in treatment of smear positive pulmonary tuberculosis (PTB) patients with background HIV infection. PMID- 15819475 TI - Symptomatic depression after long-term steroid treatment: a case report. AB - Good results of the treatment of mood disturbances and psychoses induced with steroid administration are in agreement with the fact described in literature of serotonin release reduction caused by steroids and possible relationship between depressive and psychotic manifestations and low seretonin level in cerebrospinal fluid. The patient had no head trauma in the past. He denied any loss of consciousness. The patient at the age of six years had type C virus hepatitis which changed into chronic hepatitis with a slight autoimmune component. He received six months oral steroid (prednisone) treatment in 20 mg daily dose resulting in symptomatic depression. The patient was treated by the author with sertraline from low doses up to 150 mg daily. A remission of the depressive illness and OCD was obtained. The author decided to treat the patient with sertraline in view of its high safety in somatic diseases and good tolerance. PMID- 15819476 TI - Chronic pyonephrosis associated with renal neovascularisation. AB - The spectrum of renal tract infections is wide. When the kidney has been severely damaged, the radiological findings may suggest a malignancy. To report a case of chronic pyonephrosis, which even at exploration appeared to be a malignancy. The case record of the patient as well as the literature were reviewed and reported. A 25 year-old woman presented with a 5-year history of left lumbar pain, urinary frequency and intermittent total haematuria. The intravenous urography showed non function in the left kidney harbouring a calculus. Treatment was delayed for poor finances. At laparotomy a huge renal mass invading the colonic mesentery and showing neovascularisation was removed. The final diagnosis was chronic pyonephrosis. She recovered from postoperative septicaemia. Neovascularisation is a feature of malignant disease mediated by angiogenesis factors. These factors are probably present in chronic inflammation. It is suggested that for nephrectomy, prophylactic antibiotics should be used. There is also a need for histopathological examination of every specimen removed at operation. PMID- 15819477 TI - Pregnancy after heart valve replacement--case report. AB - There is an increased risk of thromboembolism, anticoagulant-related haemorrhage, foetal-wastage, and congestive cardiac failure in pregnant women with mechanical heart valves. Pregnancy in such patients is a high risk venture. In order to have a good outcome, the care of such patients must necessarily be multidisciplinary and in a well-equipped centre with adequate support services. One such patient who had mechanical mitral valve replacement in 1986 and was on warfarin anticoagulant presented in April 2003 with a first trimester pregnancy. She was in stable haemodynamic state and went through pregnancy without event. Delivery was by an elective caesarian section at 38 weeks gestation. The care of this patient during pregnancy, delivery, and puerperium is the basis of this report. PMID- 15819478 TI - A review of incidents of rape from police records in Ibadan, Nigeria. PMID- 15819479 TI - Design aspects of compliant, soft layer bearings for an experimental hip prosthesis. AB - Currently, an artificial hip joint can be expected to last, on average, in excess of 15 years with failure due, in the majority of cases, to late aseptic loosening of the acetabular component. A realistic alternative to the problem of wear in conventional joints is the introduction of bearing surfaces that exhibit low wear and operate in the full fluid-film lubrication regime. Contact analyses and friction tests were performed on compliant layer joints (metal-on-polyurethane) and the design of a prototype ovine arthroplasty model was investigated. When optimized, these components have been shown to achieve full fluid-film lubrication. PMID- 15819480 TI - Analysis of the contact interactions between fingertips and objects with different surface curvatures. AB - Previous experimental observations indicated that the contact interactions between finger and tool handle interfere with the grasp stability, affecting the comfort and manipulations of handheld tools. From a biomechanical point of view, the curvature of the contact surface should affect the contact pressure and contact area, and thereby the comfort and manipulations of hand tools. The current authors analysed, via a finite element model, the contact interactions between fingertips and objects with different curvatures. The effects of the curvature on the contact stiffness, fingertip deformations, contact pressure distributions, and stress/strain distributions within the soft tissues were analysed. The simulation results indicated that the curvature of the contact interface influences the contact characteristics significantly. For a given contact force, the contact area and the contact stiffness increase but the contact pressure and the fingertip deformation decrease with the decrease of the contact surface curvature. The present simulation results will be useful for ergonomic designers in their aim to improve the design of tool handles. PMID- 15819481 TI - NeuFlex metacarpophalangeal prostheses tested in vitro. AB - This paper describes the testing of three single-piece silicone NeuFlex metacarpophalangeal prostheses in a finger function simulator and describes the resulting modes of prosthesis failure. In all cases, failure was due to imminent fracture of the prosthesis across the pivot of the central hinge section. This result is in contrast with previous in-vitro and in-vivo experience with single piece silicone Swanson and Sutter metacarpophalangeal prostheses, which both tend to fracture at the junction of the distal stem and the hinge. In comparison with earlier in-vitro simulator tests of the Sutter metacarpophalangeal prosthesis, the NeuFlex prostheses showed a greater longevity before fracture. To date, no other reports of fracture of the NeuFlex metacarpophalangeal prosthesis have been reported, either in vitro or in vivo. PMID- 15819482 TI - Personalised image-based templates for intra-operative guidance. AB - The high rate of unplanned perforation, poor fixation, and nerve injury with freehand pedicle screw insertion has led to the use of image-guided navigation systems. Although these improve accuracy, they have several drawbacks that could be overcome by using image-based drilling guide templates. The accuracy of such templates was tested in a cadaveric study of screw placement in the lumbar, thoracic, and cervical regions of the spine. The dimensional stability with autoclaving of duraform polyamide, to be used for manufacturing the guides, was first determined using test specimens. Computed tomography (CT) images were acquired of 4 cadaveric spines, and placement of 4 cervical, 32 thoracic, and 14 lumbar screws was planned. Eighteen personalized drilling guide templates, in four different designs, were built. Orthopaedic surgeons experienced in the freehand techniques used the templates. CT images were acquired to assess placement position with respect to the pedicle. Duraform polyamide was found to be unaffected by sterilization. Two of the template designs facilitated the placement of 20/20 screws without error. Templates can lead to successful screw placement, even in small pedicles, providing their design is optimized for the application area, e.g. with enhanced rotational stabilization. PMID- 15819483 TI - Microstrain fields for cortical bone in uniaxial tension: optical analysis method. AB - This study employed an optical strain measurement method, called microdisplacements by machine vision photogrammetry (DISMAP), to measure both the global and local strain fields in microtensile specimens of cortical bone subjected to controlled uniaxial tension. The variation of local maximum principal strains was measured within the gauge region of samples as a function of applied tensile stress during testing. High gradients of local strain appeared around microstructural features in stressed bone even while the global strain for the entire gauge region showed a strong linear correlation with increasing tensile stress (r2 = 0.98, p < 0.0001). The highest local strain around microstructural features in bone was 11.5-79.5 times higher than the global strain. PMID- 15819484 TI - A three-dimensional active shape model for the detection of anatomical landmarks on the back surface. AB - In this study relations between anatomical landmarks on the dorsal surface of the human torso corresponding to underlying skeletal structures are established. By examining the statistics of the positions of the landmarks in a training set of subjects a point distribution model is derived. Rotations of the pelvis are simulated in order to show that the main mode shapes of variation are consistent with rotations of the pelvis relative to the trunk. The parameters of these mode shapes can therefore be used as independent measures of clinical parameters such as pelvic inclination, pelvic tilt, etc. The point distribution model is further applied to improve reliability and robustness for an automatic and objective detection of the anatomical landmarks on the back surface (active shape model). The results show that it is possible to replace radiographs by surface measurements in order to measure position and orientation of the pelvis, which is particularly valuable in the case of functional examinations that normally involve a large number of radiographs (e.g. to measure the position of the pelvis in a scoliosis). PMID- 15819485 TI - Leaflet geometry extraction and parametric representation of a pericardial artificial heart valve. AB - Reverse engineering technology was used to reconstruct the complex leaflet geometry of a commercial pericardial valve in our study. Results show that the three-dimensional computer-aided design model of the leaflet surface can be rendered by fitting the surface either to cloud points or by a group of B-splines fitted to a set of cloud points that had been obtained by the process of laser scanning digitizing. However, an acceptable smooth surface is usually not guaranteed and additional manipulation is required. An alternative method is introduced in this paper, which involves the fitting of an equation to the leaflet geometry to create a smooth surface. The geometrical profile of a pericardial artificial heart valve was scanned using a laser digitizing system. The leaflet profile is represented as a set of cloud points. A quadric surface is fitted to a set of unique points, which were located on the set of cloud points. A mathematical equation is obtained by solving a least-squares fit. The geometry of the fitted leaflet surface has been proven to be closely represented by an elliptical hyperboloid. The quadratic equations of the leaflet curvatures, calculated along both the circumferential and the radial directions, resulted in simple hyperbolic curvatures. The advantages of using elliptical hyperboloid geometry for the leaflet surface are discussed and compared with other types of conicoid geometries. The concepts of parametric representation of the leaflet geometry and parametric design for leaflets are discussed. A smooth surface without inflection points and with an adjustable surface area suitable for a series of stent sizes with incremented diameters is created by this method of a single parametric design. Finally, a generic method to apply the geometry extraction and parametric representation to most pericardial heart valve prostheses was discussed. The application to valves with natural shape was introduced, challenges were identified, and a technical solution was proposed. PMID- 15819486 TI - Aerobiology in the operating room and its implications for working standards. AB - Two novel operating room (OR) ventilation concepts, i.e. the upward displacement or thermal convection system and the exponential ultra-clean laminar air flow (LAF) designed to function without extra walls, were evaluated from a bacteriological point of view. The thermal convection system (17 air changes/h) was compared with conventional ventilation (16 air changes/h) with an air inlet at the ceiling and evacuation at floor level. The exponential LAF was compared with the vertical ultra-clean LAF and the horizontal ultra-clean LAF, both with extra side walls. The comparison was made using strictly standardized simulated operations and, except for the horizontal LAF, it was performed in the same OR where the type of ventilation was changed. In the different areas important for surgical asepsis, the thermal system resulted in a twofold to threefold increase in bacterial air and surface counts compared to the conventional system (statistical significance = p < 0.05-0.0001). The bacteriological efficiency of the exponential LAF was equal to the horizontal and vertical LAF units with extra walls in the OR, and all three systems easily fulfilled the criteria for ultra clean air, i.e. bacteria-carrying particles < 10/m3. In the areas important for surgical asepsis the turbulent ventilation systems yielded highly significant correlation between air and surface contamination (p < 0.02-0.0006). No such correlation existed in the LAF systems. PMID- 15819487 TI - Helen Gibbs a voice from young nursing. PMID- 15819488 TI - Managing violent patients. PMID- 15819489 TI - Time for change. PMID- 15819490 TI - The national service framework for long-term conditions. AB - The new national service framework sets quality criteria for the care of patients with long-term neurological conditions. It aims to ensure early recognition of neurological conditions, increase self-management and independent living, and improve palliative care and support for families and carers. PMID- 15819491 TI - Gonorrhoea. PMID- 15819492 TI - The positive effect of exercise as a therapy for clinical depression. AB - The positive effect of exercise on physical health has been given much attention. Research has shown it is also beneficial for mental health, and there is increasing evidence to suggest it could be used as an alternative therapy to antidepressants and psychotherapy. PMID- 15819493 TI - Nurse practitioner-led consent in day case cataract surgery. AB - This article explores issues relating to non-medical personnel gaining informed patient consent for procedures in nurse-led clinics. It examines the results of two snap-shot audits, which indicate that nurse-led consent may be effective in increasing the efficiency of clinics, while also bringing about benefits in terms of a more streamlined service for patients, and increased job satisfaction for nurses. PMID- 15819494 TI - Docusate sodium. PMID- 15819495 TI - Five steps for sustaining effective patient partnership working. AB - NHS organisations are to recruit and involve patients in planning and delivering health care services. This paper gives practical examples of recruiting and involving patients in this process and a 'five-step cycle' for sustaining effective partnership working between patients and NHS organisations. PMID- 15819496 TI - Improving oral health care of patients who have cancer. PMID- 15819497 TI - How to support students effectively. PMID- 15819498 TI - UK nurses are in demand overseas. PMID- 15819499 TI - It's a good time to make political demands. PMID- 15819500 TI - Ischemic mitral regurgitation: more knowledge, less fear, better care. PMID- 15819501 TI - Frequent mistakes in the statistical inference of biomedical data. AB - Although there is a plethora of books, reviews and articles defining the meaning of the p value, many investigators make errors when reaching conclusions from their work based on this p value. Most report their data using rigid sentences such as "the results are not statistically significant, p > 0.05", frequently misunderstood by the authors and the readers. Many professionals are not aware of their limitations in this field rendering the problem even more complicated. In this article we include advice from experts against mistakes frequently made by investigators, such as the plea made by Rothman that a correct interpretation of the data should not be replaced by sentences such as "statistically significant" or "statistically non-significant". Detailed comments on the more frequent mistakes as well as the reasons for their appearance and persistence during the decades are presented. Finally, a comprehensive explanation of the p value is included, to emphasize that to avoid these mistakes there is no need to learn the mathematical basis of tests but that logic alone would suffice. PMID- 15819502 TI - Assessment of dyssynchrony in patients with severe heart failure by nuclear imaging: paradise lost and regained or lost and gone forever? AB - Cardiac resynchronization therapy, based on biventricular and/or left ventricular preexcitation, is a recently introduced therapeutic option for patients with severe heart failure and intraventricular conduction disturbances. The invasive nature and expense of resynchronization therapy has highlighted the need to prospectively identify optimal candidates, because of the poor correlation of QRS duration with patient response. Scintigraphy and positron emission tomography made it possible the research investigation of the pathophysiological consequences of cardiac conduction disturbances on myocardial contraction, metabolism, and perfusion. Increasing evidence shows that nuclear imaging techniques allow a comprehensive evaluation of the candidates to resynchronization. In fact, phase analysis of equilibrium radionuclide angiography enables a simple, quick and reliable measurement of both of inter- and intraventricular mechanical dyssynchrony, affording an optimal predictive accuracy of the response. In addition, being scintigraphic data highly reproducible, they are suitable for sequential longitudinal follow-up of the ventricular performance and mechanical dyssynchrony in patients implanted with devices. PMID- 15819504 TI - Progression of aortic valve sclerosis and aortic valve stenosis: what is the role of statin treatment? AB - BACKGROUND: It has recently been suggested that statins could slow the progression of aortic stenosis, but this hypothesis has not been validated in large series. Moreover, there is little information about the role of statin treatment in patients with aortic valve sclerosis. METHODS: From our database 1988--2002, we retrospectively identified 1136 consecutive patients with aortic valve sclerosis (peak aortic velocity [Vmax] > 1.5 and < 2 m/s), or mild to moderate aortic stenosis (Vmax 2.0-3.9 m/s) and with > or = 2 echocardiographic studies > or = 6 months apart; 121 (11 %) were treated with statins. As a control group we randomly selected 121 age-gender-matched patients not treated with statins, with similar initial Vmax. RESULTS: The mean follow-up duration was 54+/ 34 months in the statin group, and 50+/-33 months in controls (p = 0.35). There were no differences between statin-treated patients and controls with respect to age, gender, and prevalence of hypertension. More patients in the statin group had documented hypercholesterolemia, diabetes, or had proven coronary artery disease. Overall, the rate of change of Vmax was not different between statin treated patients and controls (0.13+/-0.24 vs 0.14+/-0.19 m/s/year, p = 0.72). However, in the subgroup of patients with aortic valve sclerosis (n = 52, 26 statin-treated, 26 controls), the rate of change of Vmax was significantly lower in statin-treated patients (0.04+/-0.04 vs 0.08+/-0.06 m/s/year, p = 0.007). CONCLUSIONS: The results of our retrospective study show that statins could be beneficial in retarding the progression of valvular aortic sclerosis to aortic stenosis. This suggests that statins retard the progression of aortic valve lesion in its early stage, a finding that may have important implications in the management of this very common disease. PMID- 15819503 TI - Drug therapy during percutaneous coronary interventions in stable and unstable coronary artery disease: the Italian Drug Evaluation in Angioplasty (IDEA) study. AB - BACKGROUND: Although periprocedural drug therapy has been shown to improve the outcome of percutaneous coronary intervention (PCI), information regarding its use in daily clinical practice is limited. METHODS: We conducted a national survey on periprocedural drug therapy across the spectrum of PCI practice in Italy. Seventy-nine centers (41% of the Italian interventional cath labs) with a fair distribution across the country volunteered to enroll consecutive patients undergoing PCI for any indication from September 15 to 29, 2003. RESULTS: Of the 1517 patients enrolled, 745 (49 %) had stable coronary disease and 772 (51%) acute coronary syndromes (ACS): 457 without and 315 with ST-segment elevation. Stenting was used in 89% of cases. N-acetylcysteine was used in 23% of the patients with preexisting renal dysfunction. Thienopyridine (63% clopidogrel) pretreatment was given in 49 % of the cases and, at logistic regression analysis, was independently associated with prior myocardial infarction (p < 0.001), prior PCI (p = 0.007), stable coronary disease (p = 0.005), and treatment in northern Italy (p < 0.05). Platelet glycoprotein (GP) IIb/IIIa receptor blockers (50% abciximab 50% tirofiban) were used in 22% of the stable patients and 40 % of those with ACS, a proportion increasing to 62 % when PCI was undertaken as an emergency procedure. Off-label use of these drugs was frequent (direct cath lab use of tirofiban in 55% of the cases; bailout use: 16% with abciximab and 26% with tirofiban). At logistic regression analysis, independent predictors of GP IIb/IIIa receptor blocker use were emergency procedure (odds ratio 3.6, 95 % confidence interval 2.6 to 5.0, p < 0.0001) and treatment for an ACS (odds ratio 1.6, 95% confidence interval 1.3 to 2.1, p = 0.0002). An emergency procedure was the only independent predictor for the use of abciximab instead of tirofiban (odds ratio 4.1, 95% confidence interval 2.6 to 6.5, p < 0.0001). Triple periprocedural antiplatelet therapy, including aspirin, a thienopyridine and a GP IIb/IIIa receptor blocker was administered in only 21% of cases. At discharge, all stented patients received aspirin and a thienopyridine. Despite complete procedural success in > 90% of cases, 50% of the patients were discharged on symptomatic anti-ischemic therapy. CONCLUSIONS: A wide gap exists between guideline recommendations and periprocedural drug therapy in PCI, the only exception being full prescription of aspirin and a thienopyridine at discharge after stenting. In patients with ACS, thienopyridine pretreatment is often used as a surrogate for GP IIb/IIIa blockade, whose use rather is associated with emergency procedures. Off-label use of drugs is not uncommon. PMID- 15819505 TI - Correlations between clinical presentation, brain natriuretic peptide, big endothelin-1, tumor necrosis factor-alpha and cardiac troponins in heart failure patients. AB - BACKGROUND: The term "biochemical marker" of heart failure is used to define a biochemical substance whose plasma levels correlate with the clinical and hemodynamic status and predict the prognosis of patients with heart failure. The aim of this study was to prospectively evaluate, in a single population of patients with heart failure, the correlations between the plasma levels of brain natriuretic peptide (BNP), big endothelin-1 (BET-1), tumor necrosis factor-alpha (TNF-alpha), cardiac troponin I (cTnI) and T (cTnT), the clinical presentation, and the left ventricular function. METHODS: The study population included a series of 120 patients (97 males, 81%, mean age 56+/-12 years) in NYHA functional class I (49%), II (20%), III (26%), IV (5%) who were admitted to our institution or followed up as outpatients. All patients underwent cardiologic evaluation, standard electrocardiography, two-dimensional echocardiography, and venous blood sampling on the same day. RESULTS: At univariate analysis the following correlations were found to be significant: all the laboratory parameters correlated with the NYHA class (BNP r = 0.63, BET-1 r = 0.56, cTnI r = 0.25, cTnT r = 0.24, TNF-alpha r = 0.23); BNP (r = -0.39) and BET-1 (r = -0.27) with left ventricular ejection fraction; BNP (r = 0.37) and BET-1 (r = 0.21) with the degree of mitral insufficiency; BNP (r = -0.39), BET-1 (r = 0.25) and TNF-alpha (r = -0.19) with systolic blood pressure; cTnT (r = 0.34), cTnI (r = 0.33), BNP (r = 0.22) and BET-1 (r = 0.19) with heart rate; BNP with age (r = 0.33) and body mass index (r = -0.28). The plasma levels of BNP, BET-1, cTnT and cTnI were significantly higher in case of systemic or pulmonary congestion. At multiple regression analysis the following correlations were still present: BNP with the NYHA functional class (p < 0.005) and with pulmonary venous congestion (p < 0.05); BET-1 with the presence of pulmonary venous congestion (p < 0.005); TNF alpha with the NYHA class (p < 0.05) and systolic blood pressure (p < 0.001); cardiac troponins with heart rate (p < 0.05). CONCLUSIONS: The plasma concentrations of BNP and BET-1 showed the best and comparable correlations with parameters describing the clinical status of patients with heart failure, in particular with the presence of pulmonary venous congestion. The value of the plasma concentration of TNF-alpha and those of cardiac troponins were found to be limited in patients with relatively stable heart failure. PMID- 15819506 TI - Contrast-enhanced magnetic resonance imaging assessment of scar size in patients with chronic myocardial infarction. AB - BACKGROUND: In the assessment of myocardial infarction (MI) mass, contrast enhanced magnetic resonance imaging (CE-MRI) is comparable to single-photon emission computed tomography (SPECT). The aim of the present study was to determine whether the MI area, as assessed at CE-MRI and SPECT, is comparable to mass evaluation. We also compared CE-MRI and SPECT estimates of the MI area with functional evaluations made at echocardiography and kinetic MRI (cine-MRI). METHODS: We used a 1.0 Tesla MRI scanner and an inversion-recovery turboFLASH sequence, a tomographic gamma-camera and second-harmonic ultrasound systems. Two blinded operators assessed the extent of scarring, expressed as a percentage of the whole left ventricle (LV), using a 16-segment model. We studied 55 consecutive patients with a clinically stable healed MI (50 Q wave, 5 non-Q wave). RESULTS: The scar mass was 19+/-23% of the LV at CE-MRI and 21+/-25% at SPECT; the scar area was 29+/-23% of the LV at CE-MRI, 41+/-28% at SPECT, 29+/ 31% at cine-MRI, and 32+/-29% at echocardiography. The Bland-Altman bias between CE-MRI and SPECT mass estimations was -2% of the LV with a+/-23% limit of agreement (LOA), while the bias between the area assessments was -12% with a+/ 42% LOA. Bias between CE-MRI and functional evaluation by cine-MRI and echocardiography was 0% with a+/-39% LOA and -3% with a+/-36% LOA respectively. Comparing SPECT with cine-MRI and echocardiography the bias was 12% with a+/-52% LOA and 9% with a+/-56% LOA respectively. CONCLUSIONS: CE-MRI has proved to be comparable to SPECT in the assessment of the healed MI mass. Conversely, a high systematic error (high bias and LOA) renders CE-MRI and SPECT assessments of the MI area incomparable. Similarly (high bias and/or LOA) CE-MRI and SPECT estimations of the MI area cannot be compared with functional evaluation by echocardiography or cine-MRI. PMID- 15819507 TI - Angiographic patterns of in-stent restenosis in men and women. AB - BACKGROUND: The angiographic classification of in-stent restenosis (ISR) has been described. However, no comparative analysis of the morphologic ISR patterns between sexes has been performed. We sought to assess the ISR rate and the morphology of the different ISR patterns in the two genders. METHODS: The study population comprised 487 patients (231 male, 256 female) who underwent systematic angiographic follow-up 6 months after coronary stent implantation. RESULTS: The angiographic patterns of ISR were defined as: a) focal (length < or = 10 mm), b) diffuse (length > 10 mm and within the stent margins), c) proliferative (length > 10 mm extending beyond the stent margins), and d) totally occlusive. Angiographic ISR had a significantly higher incidence in women as compared to men (35.9 vs 29.4%, p = 0.04). In addition, angiographic analysis showed that, although all ISR patterns were present in both sexes, more males had focal ISR (59.7 vs 28.2%, p < 0.001). Conversely, females showed a higher incidence of diffuse ISR (71.8 vs 40.3%, p < 0.001), including intrastent, proliferative and occlusive patterns. The only predictors of a specific ISR pattern were diabetes mellitus and hypertension for diffuse ISR in both sexes. CONCLUSIONS: Females seem to be at higher risk of angiographic restenosis and higher levels of ISR class after coronary stent implantation. These findings may be prognostically important and should be considered for the appropriate use of newer revascularization strategies. PMID- 15819508 TI - Left ventricular aneurysm repair: early survival. AB - BACKGROUND: The aim of this study was to evaluate the early survival in patients submitted to left ventricular (LV) repair and concomitant myocardial revascularization. METHODS: We retrospectively reviewed the records of 51 patients who were submitted to LV repair and concomitant myocardial revascularization between January 1998 and June 2003. Of 51 patients (44 males with a mean age of 60+/-9.2 years, and 7 females with a mean age of 61+/-6.5 years), 29 (56.9 %) were submitted to the McCarthy technique, 16 (31.3 %) to the technique that was described by Jatene and modified by Dor, and 6 (11.8%) to the Cooley technique (linear repair). The mean preoperative LV ejection fraction was 36.5+/-7.7 %, the mean preoperative LV end-diastolic diameter was 61.8+/-3.9 mm, the mean preoperative LV end-systolic diameter was 49.9+/-5.1 mm, the mean preoperative interventricular septal thickness was 9.7+/-1.7 mm, and finally, the mean posterior wall thickness was 8.9+/-1 mm. The mean follow-up was 30.7+/-23.4 months (range 11-82 months). RESULTS: One patient died during surgery (1.9%) and one early postoperatively (1.9%). The causes of death were respectively irreversible ventricular fibrillation and low cardiac output syndrome. The overall survival at follow-up was 98% (49 patients). One patient died during follow-up of myocardial infarction. At follow-up, all patients presented with improved clinical symptoms, and had a better mean NYHA functional class with respect to the preoperative value (3.3+/-0.3 vs 2.0+/-0.5, p < 0.05). Besides, the mean CCS angina class decreased in all patients (3.4+/-0.2 vs 1.9+/-0.3, p < 0.05). The average LV ejection fraction increased from 36.3+/-7.7 to 44.3+/-4.9% (p < 0.001), the average LV end-diastolic diameter decreased from 61.7+/-3.9 to 55.5+/-5.6 mm (p < 0.001), and the average LV end-systolic diameter decreased from 49.9+/-5.1 to 40.4+/-5.1 mm (p < 0.001). No statistically significant difference was found between the preoperative and postoperative data regarding the interventricular septal thickness (9.7+/-1.7 vs 10.3+/-1.6 mm, p = NS), and the posterior wall thickness (9.7+/-1 vs 8.8+/-1.3 mm, p = NS). CONCLUSIONS: LV aneurysm repair and concomitant myocardial revascularization may be performed with an acceptable surgical risk and a good early survival. PMID- 15819509 TI - Pulmonary artery stenting without angiographic imaging. AB - Pulmonary artery stenosis is a frequent complication seen after surgical repair of tetralogy of Fallot. In this setting, endovascular stent implantation is now accepted as the first-choice therapeutic option. However, angiographic imaging still being held as mandatory to check the stent position before final deployment, this procedure is not considered suitable for patients who cannot be submitted to angiography. In this paper, we report a novel method for the correct implantation of an endovascular stent without angiographic imaging. A 9-year-old boy underwent cardiac catheterization to relieve a severe left pulmonary artery stenosis. A previous attempt had been aborted due to a life-threatening anaphylactic reaction to the contrast medium. To avoid angiography, a contrast medium-filled compliant atrial septal defect sizing balloon (Amplatzer Sizing Balloon, AGA Medical Corporation) was used to image the vessel stenosis and successfully guide stent deployment. After the procedure, the transstenotic pressure gradient disappeared and the left-to-right pulmonary perfusion imbalance almost completely reverted. PMID- 15819510 TI - Usefulness of transthoracic color Doppler echocardiography for the identification of left anterior descending coronary artery stenosis. AB - We briefly describe our experience with transthoracic Doppler echocardiography for the direct evaluation of mid-distal left anterior descending coronary artery (LAD) stenosis. Three patients with previous myocardial infarction, scheduled for coronary flow reserve evaluation, underwent Doppler analysis of the mid-distal LAD. In all 3 cases, the mid-distal LAD stenosis was accurately quantified by the Doppler spectrum as confirmed by quantitative coronary angiography. Our study demonstrated the feasibility of transthoracic Doppler echocardiography in the discrimination of significant and non-significant mid-distal LAD stenosis. Limitations of such a technique could be related to the variable coronary anatomy and to the severity of the atherosclerotic process. PMID- 15819511 TI - Quadricuspid aortic valve as a cause of severe aortic regurgitation. AB - Quadricuspid aortic valves (QAVs) constitute a rare congenital malformation, with an incidence ranging from 0.008 to 0.048%. We report a case of severe aortic regurgitation associated with a QAV, which was diagnosed intraoperatively using transesophageal echocardiography. Since the first case described in 1862, 186 QAVs have been reported. In most cases, QAVs are associated with valve regurgitation, with a concurrent stenosis in some patients, while only a small number of QAVs are functionally normal. Once the diagnosis has been made, echocardiographic follow-up is recommended, as progression to severe valve regurgitation is common. Antibiotic prophylaxis is advisable for dental, and "dirty" surgical procedures, to minimize the risk of infective endocarditis. PMID- 15819512 TI - Unusual tumor-like calcification of the mitral annulus: diagnosis and tissue characterization by ultrasound, computed tomography and magnetic resonance imaging. AB - Calcification of the mitral annulus is a common echocardiographic finding in the elderly, particularly in females. Calcium deposits are generally located in the posterior mitral ring, sometimes extending to the whole mitral annulus and involving the mitral valve apparatus. The present report refers to 2 patients with a very atypical mass-like calcification of the mitral annulus resembling a cardiac tumor. A detailed evaluation of the mass was obtained at transthoracic and transesophageal echocardiography; the differential diagnosis with other intracardiac masses was aided by the use of computed tomography and magnetic resonance imaging. To our knowledge there has been no prior report of such a lesion evaluated at cardiac magnetic resonance imaging. PMID- 15819513 TI - Platypnea-orthodeoxia induced by fenestrated atrial septal aneurysm. AB - Platypnea-orthodeoxia is a peculiar syndrome characterized by a right-to-left shunt, which occurs in the upright position. The diagnosis is made by contrast transesophageal echocardiography, paying attention to include contrast visualization in the orthostatic decubitus. The association of this syndrome with a fenestrated atrial septal aneurysm is rare and probably underlies a peculiar and also rare mechanism of shunting in presence of normal pulmonary pressure. We report of a case of a 58-year-old man with a fenestrated atrial septal aneurysm and platypnea-orthodeoxia syndrome treated by surgical closure of the atrial defect. PMID- 15819514 TI - [Orthotopic murine model of head and neck squamous cell carcinoma]. AB - PURPOSE: To develop a reliable animal model able to reproduce the behavior of head and neck squamous cell carcinomas (HNSCC). This model should facilitate our understanding of the molecular mechanisms of tumorigenicity and progression of these tumors, as well as the evaluation of novel therapies. MATERIAL AND METHODS: 20 nude mice nu/nu were injected intraorally and submucosally with a cell line derived from a human squamous cell carcinoma of the glottis. RESULTS: 90% of the mice developed locally agressive squamous cell carcinomas, invading the surrounding muscle fibers and into loose connective tissue structures. All the tumors showed perineural growth. Four (22%) of the 18 mice showed bone destruction, and 22% vascular invasion. Tumor cells invaded lymphatic vessels in all the specimens, and 100% of the mice developed regional lymph node metastases. None of the animals developed haematogenous metastases. CONCLUSIONS: We present a metastasing model of HNSCC that resembles its human counterpart in many aspects. PMID- 15819515 TI - [Controlling retests in a universal hearing screening program]. AB - INTRODUCTION: To evaluate the influence of several factors in the need for retesting in the first stage of a universal auditory screening program, a prospective and statistic study is presented, specially remarking the differences found between two health systems (public and private). PATIENTS AND METHODS: 18,073 children born in the 1999-2004 period were included in the study, in the context of a universal screening protocol based on Otoacoustic emissions and ABR, and distributed into three groups depending on their place of birth (A: public hospital; B: private hospital; C: children from other region). RESULTS: Significant differences were found between the groups (7.3% retesting in the public system and 2.4% in the private), explained by the fact that in the first group the test was performed in 74.9% of cases within 3 days after birth (when the need for repetition was 7,7%), and in the other groups the exploration was delayed up to 4-10 days in most cases (when retesting was performed only in 4.9%). DISCUSSION AND CONCLUSIONS: Proportion of retesting in the first stage of a universal screening program is an important factor for the cost estimation of these protocols. We assume that the age at testing varies significantly this factor, and on the contrary, we know that precocious exploration (before the child leaves the hospital) increases the program covering. Therefore we conclude that the test should be delayed only in health systems needing to save resources or not able to test before discharge, and not in those wanting to guarantee the maximum covering. PMID- 15819516 TI - [Kinetic tests representativity in vestibular function]. AB - INTRODUCTION: The most frecuently tests used to study the vestibular system, with videonystagmographic register, are the caloric and rotatory ones. MATERIAL AND METHODS: We have carried out a comparative study of the kinetic tests through a sample of 41 patients without vestibular pathology, stratified by age and gender. The performed rotational tests have been: Constant/cycles, postrrotatory test, increasing, decreasing and constant test. RESULTS: We have not observed age or gender influence on the results. We give the numeric results of these tests. CONCLUSION: The obtained data allow to conclude that the information given by the increasing and decreasing pendulous test is similar, phenomenon that doesn't happen in the rest of the practiced tests. We also study the habituation phenomenon to a know stimuli. We point out the importance of each laboratory to have their tests done previously in a normal control group in order to compare the results when performing them in pathological patients. PMID- 15819517 TI - [Postural analysis of the test "timed-up-and-go" in patients with vertigo]. AB - INTRODUCTION AND OBJECTIVES: To characterize and analyze the "Timed-up-and-go" test in non-fallers patients with peripheral vertigo. To assess if turning to the side of the lesion induces a more intense instability. MATERIAL AND METHODS: The test was performed after carefully explanation to the patient who was allowed to do some attempts. Postural measurements were the angle and angular speed of trunk sway in roll and pitch. It was done with a special device adapted to the trunk with two sensors specially built for that purpose. CONCLUSIONS: Dizzy, non fallers patients perform this test in a period of time considered as normal. Canal paresis influences in roll trunk sway when the patient performs the TUG only when turning to the normal side. PMID- 15819518 TI - [Sublingual immunotherapy in seasonal allergic rhinitis. Review of 30 cases]. AB - INTRODUCTION: Specific immunotherapy consists in administering increasing doses of antigen, to which the patient is sensitive, to determine a protective immune response. PATIENTS AND METHODS: A descriptive and retrospective review of 30 patients diagnosed of seasonal allergic rhinitis, all treated with specific immunotherapy. Data on age, gender, symptoms, prick test before and after the immunotherapy administration, drugs, efficacy and side effects, were recorded using the SPSS statistical program. RESULTS: 90% had moderated symptoms and 10% had severe symptoms and asthma. 33% were sensitized to grass pollen. 60% needed antihistamines and intranasal corticosteroids before the immunotherapy. In a lot of cases (80%) there were no side effects. The complete desensitization occurred in 53.3% and the reduction of symptoms in 73% of the cases. 54% of the patients didn't need treatment after specific immunotherapy. CONCLUSIONS: The specific immunotherapy is the only curative therapy. Sublingual therapy is one of the most accepted because is well tolerated, it uses a higher allergen concentration, it reduces the symptoms and the needs of drugs and side effects are uncommon. PMID- 15819519 TI - [Development of an olfactory screening test based on the Connecticut test (CCCRC)]. AB - OBJECTIVE: To create a screening olfactory test based on the CCCRC (Connecticut Chemosen-sory Clinical Research Center). MATERIAL AND METHODS: We compare the screening test based on CCCRC with PST (Pocket Smell Test) based on UPSIT in 40 patients with nasal poliposis, in order to de-termine the specificity, sensitivity, positive predictive and negative predictive value. The validity index was 95% and accuracy rate was 10%. We determine unit cost, the time required to perform the test in outpatients office and how difficult it is to do the test. RESULTS: Sensibility was 88%, specificity was 77%. The positive predictive value was 34% and the negative predictive value was 94%. The unitary cost of our screening test was 0.57 euros when it is performed by a nurse. The unitary cost of PST is 1.76 euros. To perform our screening test took 2.8 +/- 0.4 minutes. The 96% of the patients thought the test was easy to do. CONCLUSION: Our test is a valid screening test to be used in patients with nasal poliposis. PMID- 15819520 TI - [Goya, his deafness and his time]. AB - We have made a bibliographyc review of the most important otology books from the end of the XVIII to the begining of the XIX century. We propose several hipothesis of the diagnoses and treatment of Goya's deafness in relation to the contemporary otological advances. PMID- 15819521 TI - [Cochlear implant in a patient with dilatation of the endolymphatic sac and vestibular aqueduct]. AB - We present an 8 years patient old with a giant dilatation of the vestibular aqueduct associated with profound and progressive bilateral sensorineural hearing loss. The patient received a cochlear implant and during the surgery a pulsatile clear fluid via the cochleostomy was observed, without gusher. We present all the problems, and offer some advices for this kind of surgery. PMID- 15819522 TI - [Papillary carcinoma of the thyroid metastatic to the parapharyngeal space]. AB - This is a case report of a papillary thyroid carcinoma metastatic to the parapharyngeal space, that presented with local obstructive symptoms (dysphagia and displacement of the ipsilateral tonsil). The diagnosis was suspected by imaging studies (CT and MRI) but not confirmed until histological examination. It is not common for such tumors to metastasize to the parapharyngeal space. This is the reason why, to our knowledge, there have been described only five similar cases previously. PMID- 15819523 TI - Etiology and management of implant complications. Interview. PMID- 15819524 TI - Ready steady... Scotland gears up for the implementation of its new Mental Health Act. PMID- 15819525 TI - Graduation day. People with long-standing severe mental illness have good reason to dread their 65th birthday. PMID- 15819526 TI - Zero tolerance. Has women's safety on psychiatric wards fallen off the government's radar? PMID- 15819527 TI - You can ring our bell. No Panic runs its comprehensive support service for people with anxiety disorders entirely by telephone. PMID- 15819528 TI - To be out about his mental health problems and his sexuality was almost unimaginable. PMID- 15819529 TI - What will they do when there's someone insisting, 'I'm fine' and the needle's flying off the scale? PMID- 15819530 TI - In the global village we in the west have much to learn from our non industrialised neighbours. PMID- 15819531 TI - Tackling the attitude problem. The achievements to date of Scotland's 'see me' anti-stigma campaign. PMID- 15819532 TI - How do we treat unhappiness? The potential for supported self-help in primary care settings. PMID- 15819533 TI - Torn up with anger. What happens to male survivors of childhood sexual abuse? PMID- 15819534 TI - Teachers don't know a lot. Who better to teach young people about mental health than young people themselves? PMID- 15819535 TI - This life. PMID- 15819536 TI - Hydrothermal synthesis of ultralong and single-crystalline Cd(OH)2 nanowires using alkali salts as mineralizers. AB - Ultralong and single-crystalline Cd(OH)(2) nanowires were fabricated by a hydrothermal method using alkali salts as mineralizers. The morphology and size of the final products strongly depend on the effects of the alkali salts (e.g., KCl, KNO(3), and K(2)SO(4) or NaCl, NaNO(3), and Na(2)SO(4)). When the salt is absent, only nanoparticles are observed in TEM images of the products. The 1D nanostructure growth method presented herein offers an excellent tool for the design of other advanced materials with anisotropic properties. In addition, the Cd(OH)(2) nanowires might act as a template or precursor that is potentially converted into 1D cadmium oxide through dehydration or into 1D nanostructures of other functional materials (e.g., CdS, CdSe). PMID- 15819537 TI - Spin density distribution in five- and six-coordinate iron(II)-porphyrin NO complexes evidenced by magnetic circular dichroism spectroscopy. AB - Using magnetic circular dichroism (MCD) spectroscopy together with DFT calculations, the spin density distributions in five-coordinate [Fe(TPP)(NO)] (I) and six-coordinate [Fe(TPP)(MI)(NO)] (II, MI = 1-methylimidazole) are defined. In the five-coordinate complex, a strong Fe-NO sigma bond between pi(*)(h) and d(z)(2) is present that leads to a large transfer of spin density from the NO ligand to Fe(II) corresponding to an electronic structure with noticeable Fe(I) NO(+) character. Consequently, the MCD spectrum is dominated by paramagnetic C term contributions. On coordination of the sixth ligand, the spin density is pushed back from the iron toward the NO ligand, resulting in an Fe(II) NO(radical) type of electronic structure. This is reflected by the fact that the MCD spectrum is dominated by diamagnetic contributions. PMID- 15819538 TI - Pb(II) coordination and synergistic ion-exchange extraction by combinations of sulfonamide chelates and 2,2'-bipyridine. AB - The disulfonamide ligands 1,2-C(6)H(4)(NH(2)SO(2)C(6)H(5))(2) (1) and 1,2 C(6)H(4)(NH(2)SO(2)C(6)H(4)-p-Bu(t))(2) (2), which are readily available in good yields from o-phenylenediamine and the corresponding sulfonyl chlorides, efficiently extract Pb(II) from water into 1,2-dichloroethane when used in synergistic combinations with 2,2'-bipyridine via an ion-exchange mechanism. The extraction was shown to proceed via the formation of a ternary Pb-sulfamido-2,2' bipyridine complex. The X-ray crystal structure of the binary Pb-sulfamido complex 3 shows a coordination polymer with a stereochemically active lone pair on Pb formed by S=O-Pb axial coordination. PMID- 15819539 TI - Preparation and solid-state characterization of the novel mixed biradical *NSNSC CNSSN*. AB - Reduction of the radical-cation [*NSSNC-CNSNS][AsF(6)] with ferrocene affords the novel biradical *NSNSC-CNSSN* containing both 1,2,3,5- and 1,3,2,4-isomeric dithiadiazolyl rings. Biradicals form centrosymmetric dimers with pi(*)-pi(*) interactions between different isomeric rings. Biradical *NSNSC-CNSSN* is diamagnetic in the solid state (C = 0.00035, TIP = 6.5 x 10(-)(5) emu/Oe.mol); however, an increase in paramagnetism was observed upon grinding (C = 0.003, TIP = 4.2 x 10(-)(4) emu/Oe.mol). PMID- 15819540 TI - High-resolution X-ray emission spectroscopy of molybdenum compounds. AB - High-resolution molybdenum K-edge X-ray emission spectroscopy (XES) was used to characterize the K beta(4) and K beta' ' valence-to-core transition bands in the oxo-Mo compounds K(2)MoO(4), MoO(S(2)CNEt(2))(2), and MoO(2)(S(2)CNEt(2))(2). The K beta(4) and K beta' ' emission bands are attributed to transitions to the Mo 1s core hole from molecular orbitals possessing primarily molybdenum 4d and oxygen 2s character, respectively. This communication describes the first assignment of the K beta' ' interatomic band in the emission spectra of molybdenum complexes. Additionally, the K beta(4) and K beta' 'transitions are shown to be sensitive to the chemical and electronic environment of the metal, suggesting that high resolution XES might be an effective method for elucidating the nature of the molybdenum centers in biological systems. PMID- 15819541 TI - Photosensitizer in a molecular bowl: steric protection enhancing the photonuclease activity of copper(II) scorpionates. AB - Steric encumbrance caused by the tripodal ligand in the ternary tris(3 phenylpyrazolyl)borate copper(II) heterocyclic base complexes [Cu(B)(Tp(Ph))](ClO(4)) (B = dipyridoquinoxaline, dipyridophenazine) leads to efficient cleavage of supercoiled DNA to its relaxed form upon exposure to red light at 632.8 and 694 nm as a result of protection of the photosensitizer in the molecular bowl of the {Cu(Tp(Ph))} moiety, which generates singlet oxygen as the reactive species in a type-II process. PMID- 15819542 TI - Self-assembly of a fluorescent chiral zinc(II) complex that leads to supramolecular helices. AB - The chiral Zn(II) complex [ZnLCl(2)], 1 {L = 4-methyl-2,6-di[(S)-(+)-1 phenylethyliminomethyl] phenol}, self-assembles via C-H...Cl hydrogen bonding into supramolecular helices. Complex 1 exhibits emission in solution at room temperature in the visible range. Crystal data for 1: orthorhombic space group P2(1)2(1)2(1), a = 9.614(2) A, b = 13.825(3) A, c = 18.667(3) A, V = 2481.1(8) A(3), Z = 4. PMID- 15819543 TI - Binding of propylene oxide to porphyrin- and salen-M(III) cations, where M = Al, Ga, Cr, and Co. AB - The binding of propylene oxide (PO) to a series of metal cations LM(III)(+), where for L = tetraphenylporphyrin (TPP) M = Al, Ga, Cr, and Co, and for L = (R,R)-N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexenediamine (salen) M = Al and Cr, was studied in the gas phase by electrospray tandem mass spectroscopy, and the relative stabilities of LM(PO)(2)(+) and LM(PO)(+) cations were determined. The chromium(III) and aluminum(III) cations most tenaciously bind PO, and for M = Al, coordination to the TPP ligated metal center was favored relative to salen. For (TPP)M(PO)(2)(+), the dissociation of PO followed the order M = Al > Cr, but for (TPP)M(PO)(+) the dissociation was M = Cr > Al. The single-crystal structural determinations on (R,R-salen)AlOCHMe(S)CH(2)Cl.0.5PO and (R,R salen)AlO(2)CMe.1.5py grown in neat PO and pyridine, respectively, reveal five coordinate aluminum(III) centers with the alkoxide/acetate ligands in the axial position of a square-based pyramid. These results are discussed in terms of the reactivity of these metal complexes in ring-opening polymerizations and copolymerizations with PO and CO(2), respectively. PMID- 15819544 TI - Mononuclear titanium(IV)-citrate complexes from aqueous solutions: pH-specific synthesis and structural and spectroscopic studies in relevance to aqueous titanium(IV)-citrate speciation. AB - Titanium is a metal frequently employed in a plethora of materials supporting medical applications. In an effort to comprehend the involvement of titanium in requisite biological interactions with physiological ligands, synthetic efforts were launched targeting aqueous soluble species of Ti(IV). To this end, aqueous reactions of TiCl(4) with citric acid afforded expediently, under pH-specific conditions, the colorless crystalline materials Na(6)[Ti(C(6)H(4.5)O(7))(2)(C(6)H(5)O(7))].16H(2)O (1) and Na(3)(NH(4))(3)[Ti(C(6)H(4.5)O(7))(2)(C(6)H(5)O(7))].9H(2)O (2). Complexes 1 and 2 were characterized by elemental analysis, FT-IR, (13)C-MAS solid state and solution NMR, cyclic voltammetry, and X-ray crystallography. 1 crystallizes in the triclinic space group P, with a = 15.511(9) A, b = 15.58(1) A, c = 9.848(5) A, alpha = 85.35(2) degrees, beta = 76.53(2) degrees, gamma = 61.97(2) degrees, V = 2042(2) A(3), and Z = 2. 2 crystallizes in the triclinic space group P, with a = 12.437(5) A, b = 12.440(5) A, c = 12.041(5) A, alpha = 83.08(2) degrees, beta = 81.43(2) degrees, gamma = 67.45(2) degrees, V = 1697(2) A(3), and Z = 2. The X ray structures of 1 and 2 reveal the presence of a mononuclear complex, with Ti(IV) coordinated to three citrate ligands in a distorted octahedral geometry around Ti(IV). The citrates employ their central alkoxide and carboxylate groups to bind Ti(V), while the terminal carboxylates stay away from the Ti(IV)O(6) core. Worth noting in 1 and 2 is the similar mode of coordination but variable degree of protonation of the bound citrates, with the locus of (de)protonation being the noncoordinating terminal carboxylates. As a result, this work suggests the presence of a number of different Ti(IV)-citrate species of the same nuclearity and coordination geometry as a function of pH. This is consistent with the so far existing pool of mononuclear Ti(IV)-citrate species and provides a logical account of the aqueous speciation in the requisite binary system. Such information is vital in trying to delineate the interactions of soluble and bioavailable Ti(IV) forms promoting biological interactions in humans. To this end, chemical properties, structural attributes, and speciation links to potential ensuing biological effects are dwelled on. PMID- 15819545 TI - Low-coordinate chromium siloxides: the "box" [Cr(mu-Cl)(mu-OSitBu3)]4, distorted trigonal [(tBu3SiO)3Cr][Na(benzene)] and [(tBu3SiO)3Cr][Na(dibenzo-18-c-6)], and trigonal (tBu3SiO)3Cr. AB - Treatment of CrCl(2)(THF)(2) with NaOSi(t)Bu(3) afforded the tetrameric "box" [Cr(mu-Cl)(mu-OSi(t)Bu(3))](4) (1, X-ray). THF cleaved 1 to provide trans (silox)ClCr(THF)(2) (2), whereas degradation of 1 with 4-picoline caused disproportionation and the generation of trans-Cl(2)Cr(4-pic)(2) and trans (silox)(2)Cr(4-pic)(x) (n = 2, 3; 3, 3-4-pic). Chromous centers in 1 were antiferromagnetically coupled, and density functional calculations on the high spin (multiplicity = 17) model [Cr(mu-Cl)(mu-OH)](4) (1') revealed that its singly occupied 3d orbitals spanned an energy range of approximately 2 eV. The addition of 8 equiv of Na(silox) to 1 yielded [((t)Bu(3)SiO)Cr(mu OSi(t)Bu(3))(2)]Na.C(6)H(6) (4, Y shaped, angle OCrO(Na) = 91.28(7) degrees), and treatment of 4 with dibenzo-18-crown-6 produced [(silox)(3)Cr][Na(dibenzo-18 crown-6)] (5, angle OCrO = approximately 120 degrees, (120 + alpha) degrees, (120 - alpha) degrees). Calculations of [((t)Bu(3)SiO)Cr(mu-OSi(t)Bu(3))(2)]Na (4') and Cr(silox)(3)(-) (5') provided reasonable matches with the experimental geometries (X-ray). The trigonal chromic derivative (silox)(3)Cr (6) was synthesized from CrCl(3)(THF)(3) for structural and calculational comparisons to the chromous derivatives. PMID- 15819546 TI - A platinum acetylide polymer with sterically demanding substituents: effect of aggregation on the triplet excited state. AB - The effect of interchain interaction on the triplet excited state is explored in two Pt-acetylide polymers of the type [-trans-Pt(PBu(3))(2)-C triple bond C-Ar-C triple bond C-](n), where Ar is either 1,4-phenylene or is based on the pentiptycene unit (polymers 2 and 3, respectively). To explore the effect of interchain interaction in Pt-acetylide materials, the optical properties of parent polymer 2 are compared with those of polymer 3 in which interchain interaction is precluded by the sterically bulky pentiptycene moiety. Insight into the effect of the pentiptycene unit on packing in the solid state comes from the X-ray structure of monomer 1b, Ph-C triple bond C-[trans-Pt(PBu(3))(2)]-C triple bond C-Ar-C triple bond C-[trans-Pt(PBu(3))(2)]-C triple bond C-Ph. Spectroscopic studies indicate that weak phosphorescence emission from an interchain aggregate is observed from parent polymer 2, both in solution and in the solid state. By contrast, the photophysics of 3 is dominated by the intrachain triplet exciton. Interestingly, the phosphorescence emission of polymer 3 in the solid state is nearly superimposable with that of a single crystal of monomer 1b, suggesting that the solid polymer experiences an environment that is similar to that of the monomer in the crystal. PMID- 15819547 TI - Facile synthesis, structure, and luminescence properties of Pt(diimine)bis(arylacetylide) chromophore-donor dyads. AB - The luminescent complex Pt(dpphen)bis(arylacetylide) complex (1) (dpphen = 4,7 diphenylphenanthroline and arylacetylide = 4-ethynylbenzaldehyde) has been synthesized and characterized structurally and spectroscopically. Complex 1 has been employed in the synthesis of donor-chromophore (D-C) dyads through Schiff base condensations of different anilines to give imine-linked dyads 2-4 and through imine reduction with borohydride, to give the corresponding amine-linked dyads 2a-4a. Crystal structure determinations of 1-4 and 4a establish a distorted square-planar geometry around the Pt(II) ion in each system with cis arylacetylide ligands and a diimine-constrained N-Pt-N bond angle of ca. 79.5 degrees. Complex 1 is strongly emissive having a relative quantum yield (phi) of 36% and an excited-state lifetime of 3.1 micros. In accord with the notion of photoinduced electron transfer from the aniline-based donor to the photoexcited chromophore, the emission of dyads 2-4 and 2a-4a is effectively quenched in all solvents tested. The intense absorption at 400 nm (30000-70000 L/mol.cm) for 2 and 2a has been assigned as an intraligand pi-pi* transition, whereas the lowest energy transitions for all other dyads correspond to Pt-to-pi(diimine) MLCT transitions. Although the dyads can be synthesized in a facile manner, photolysis experiments reveal that both the imine and amine linkages are photochemically unstable, resulting in hydrolysis and regeneration of the aldehyde-containing chromophore 1. PMID- 15819548 TI - Crystal growth and magnetic properties of lanthanide-containing osmium double perovskites, Ln2NaOsO6 (Ln = La, Pr, Nd). AB - A series of double perovskite oxides, Ln(2)NaOsO(6) (Ln = La, Pr, Nd), has been prepared as single crystals from acidic molten NaOH. All three oxides crystallize in the monoclinic space group P2(1)/n (Glazer tilt system #10, a(-)a(-)b(+)), forming a 1:1 ordered rock salt lattice of the Na(+) and Os(5+) cations. Magnetic susceptibility measurements show evidence of antiferromagnetic correlations in La(2)NaOsO(6) and of a spin-flop transition from an antiferromagnetic to ferromagnetic-like state in both Pr(2)NaOsO(6) and Nd(2)NaOsO(6). PMID- 15819549 TI - Coordination polymers from silver(I) and bifunctional pyridyl ligands. AB - Coordination polymers and a macrocycle formed from the reactions between flexible bis(2-pyridyl) ligands and AgCF(3)SO(3) are reported. The type of structure formed depends on the choice of ligand and the stoichiometry of the reaction. When 1 equiv of 1,4-bis(pyridin-2-ylmethoxy)benzene (L2), 4,4'-bis(pyridin-2 ylmethoxy)biphenyl (L4), or bis((4-pyridin-2-ylmethoxy)phenyl)methane (L5) is used, 1D chain coordination polymers held together via Ag-N bonds are generated. When a 2:1 ratio of L2 and silver ion is used, a 2D porous network is formed. The reaction between silver ions with a mixture of ligands (L1 and L2 in 1:1 ratio, L1 = 1,4-bis((pyridin-2-yl-methyl)thio)benzene) results in a novel 1D ABAB type coordination copolymer where L1 and L2 act as a bis-bidentate and a bis monodentate ligand, respectively. The reaction of 1-(pyridin-2-ylmethoxy)-4 ((pyridin-2-yl-methyl)thio)benzene (L3) with silver ions in a 1:1 ratio gives a bimetallic macrocycle rather than a polymeric species. Structural analyses of the polymeric compounds suggest that interactions between the aromatic rings play a significant role in stabilizing the polymeric structures. PMID- 15819550 TI - Ligand-free deltahedral clusters of silicon in solution: synthesis, structure, and electrochemistry of Si9(2-). AB - Deltahedral nine-atom clusters of silicon, Si(9)(2-), were synthesized by mild oxidation of a liquid ammonia solution of K(12)Si(17) with Ph(3)GeCl in the presence of 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane) or 2,2,2-crypt (4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8.8.8]hexacosane). The clusters were structurally characterized in [K(18-crown-6)](2)Si(9).C(5)H(5)N (yellow; orthorhombic, Pnma; a = 14.013(1), b = 18.108 (1), c = 18.320 (1) A; Z = 4) crystallized from a pyridine solution of the product of the aforementioned reaction in liquid ammonia. Si(9)(2-) is the first unequivocally characterized nine-atom cluster of group 14 with a charge of 2-. In addition to pyridine, the product from the reaction in liquid ammonia is also soluble in DMF, and the Si(9)(2-) clusters were characterized by mass spectrometry in such a solution. The more reduced clusters Si(9)(3-) have also been crystallized from pyridine solution. Cyclic voltammetry in both pyridine and DMF solutions clearly shows the Si(9)(2-)/Si(9)(3-) redox couple as one-electron reversible process. The structural similarities and differences between Si(9)(3-) and Si(9)(2-) are discussed herein. PMID- 15819551 TI - The satellite-shaped Co-15 Polyoxotungstate, [Co6(H2O)30{Co9Cl2(OH)3(H2O)9(beta SiW8O31)3}]5-. AB - The 15-cobalt-substituted polyoxotungstate [Co(6)(H(2)O)(30){Co(9)Cl(2)(OH)(3)(H(2)O)(9)(beta-SiW(8)O(31))(3)}](5-) (1) has been characterized by single-crystal XRD, elemental analysis, IR, electrochemistry, magnetic measurements, and EPR. Single-crystal X-ray analysis was carried out on Na(5)[Co(6)(H(2)O)(30){Co(9)Cl(2)(OH)(3)(H(2)O)(9)(beta SiW(8)O(31))(3)}].37H(2)O, which crystallizes in the hexagonal system, space group P6(3)/m, with a = 19.8754(17) A, b = 19.8754(17) A, c = 22.344(4) A, alpha= 90 degrees, beta = 90 degrees, gamma = 120 degrees, and Z = 2. The trimeric polyanion 1 has a core of nine Co(II) ions encapsulated by three unprecedented (beta-SiW(8)O(31)) fragments and two Cl(-) ligands. This central assembly {Co(9)Cl(2)(OH)(3)(H(2)O)(9)(beta-SiW(8)O(31))(3)}(17-) is surrounded by six antenna-like Co(II)(H(2)O)(5) groups resulting in the satellite-like structure 1. Synthesis of 1 is accomplished in a simple one-pot procedure by interaction of Co(II) ions with [gamma-SiW(10)O(36)](8-) in aqueous, acidic NaCl medium (pH 5.4). Polyanion 1 was studied by cyclic voltammetry as a function of pH. The current intensity of its Co(II) centers was compared with that of free Co(II) in solution. Our results suggest that 1 keeps its integrity in solution. Magnetic susceptibility results show the presence of both antiferro- and ferromagnetic coupling within the (Co(II))(9) core. A fully anisotropic Ising model has been employed to describe the exchange-coupling and yields g = 2.42 +/- 0.01, J(1) = 17.0 +/- 1.5 cm(-1), and J(2) = -13 +/- 1 cm(-(1). Variable frequency EPR studies reveal an anisotropic Kramer's doublet. PMID- 15819552 TI - Group 13 and lanthanide complexes with mixed O,S anionic ligands derived from maltol. AB - Four mixed O,S binding ligand precursors derived from maltol (3-hydroxy-2-methyl 4-pyrone) have been chelated to gallium(III), indium(III), and lanthanide(III) ions to yield a series of metal complexes. The four ligand precursors include two pyranthiones, 3-hydroxy-2-methyl-4-pyranthione, commonly known as thiomaltol (Htma), and 2-ethyl-3-hydroxy-4-pyranthione, commonly known as ethylthiomaltol (Hetma), and two pyridinethiones, 3-hydroxy-2-methyl-4(H)-pyridinethione (Hmppt) and 3-hydroxy-1,2-dimethyl-4-pyridinethione (Hdppt). Dimeric forms of the pyridinethiones, Hmppt dimer and Hdppt dimer, were also isolated and characterized. Complete characterization of the monomeric organic compounds is reported including acidity constants and crystal structures of Htma, Hetma, and Hdppt dimer. Reacting the four monomeric ligand precursors with Ga(3+) and In(3+) ions yielded new tris(bidentate ligand) complexes. X-ray-quality crystals of the fac isomer of Ga(tma)(3) were also obtained. New complexes with a range of lanthanides (Ln(3+)) were also synthesized with the two pyranthiones, Htma and Hetma. The synthesis reactions yielded complexes of the type LnL(3).xH(2)O and LnL(2)(OH).xH(2)O, as indicated by elemental analysis and spectroscopic evidence such as mass spectral data and IR and NMR spectra. PMID- 15819553 TI - Vanadium complexes with mixed O,S anionic ligands derived from maltol: synthesis, characterization, and biological studies. AB - Four mixed O,S binding bidentate ligand precursors derived from maltol (3-hydroxy 2-methyl-4-pyrone) have been chelated to vanadium to yield new bis(ligand)oxovanadium(IV) and tris(ligand)vanadium(III) complexes. The four ligand precursors include two pyranthiones, 3-hydroxy-2-methyl-4-pyranthione, commonly known as thiomaltol (Htma), and 2-ethyl-3-hydroxy-4-pyranthione, commonly known as ethylthiomaltol (Hetma), as well as two pyridinethiones, 3 hydroxy-2-methyl-4(H)-pyridinethione (Hmppt) and 3-hydroxy-1,2-dimethyl-4 pyridinethione (Hdppt). Vanadium complex formation was confirmed by elemental analysis, mass spectrometry, and IR and EPR (where possible) spectroscopies. The X-ray structure of oxobis(thiomaltolato)vanadium(IV),VO(tma)(2), was also determined; both cis and trans isomers were isolated in the same asymmetric unit. In both isomers, the two thiomaltolato ligands are arranged around the base of the square pyramid with the V=O linkage perpendicular; the vanadium atom is slightly displaced from the basal plane [V(1) = 0.656(3) A, V(2) = 0.664(2) A]. All of the new complexes were screened for insulin-enhancing effectiveness in streptozotocin-induced diabetes in rats, and VO(tma)(2) was profiled metabolically for urinary vanadium and ligand clearance by GFAAS and ESIMS, respectively. The new vanadium complexes did not lower blood glucose levels acutely, possibly because of rapid dissociation and excretion. PMID- 15819554 TI - Coordination chemistry and insulin-enhancing behavior of vanadium complexes with maltol C6H6O3 structural isomers. AB - Syntheses of vanadium complexes using the naturally occurring ligands isomaltol (Hima) and allomaltol (Hama), as well as a newly synthesized, potentially tetradentate diaminodipyrone [H(2)(en(ama)(2)], are reported. Complete characterization of the resulting compounds [trans-VO(ima)(2)(H(2)O), VO(ama)(2), V(ima)(3), V(ama)(3) and VO(en(ama)(2))], including X-ray crystallography analyses for trans-VO(ima)(2)(H(2)O) and V(ima)(3), are presented herein. Potentiometric titrations (25 degrees C, I = 0.16 M NaCl) were used to measure stability constants in the V(IV)-Hima system; these data were compared to previous data collected on the V(IV)-L (L = Hma, Hama) systems. The in vivo efficacy of these compounds to lower the blood glucose levels of STZ-diabetic rats was tested; all but VO(en(ama)(2)) produced significant decreases in plasma glucose levels. The results were compared to those of the benchmark compound BMOV [VO(ma)(2), bis(maltolato)oxovanadium(IV)], a known insulin-enhancing agent. PMID- 15819555 TI - Carbohydrate-appended 2,2'-dipicolylamine metal complexes as potential imaging agents. AB - Three discrete carbohydrate-appended 2,2'-dipicolylamine ligands were complexed to the {M(CO)(3)}(+) (M = (99m)Tc/Re) core: 2-(bis(2-pyridinylmethyl)amino)ethyl beta-d-glucopyranoside (L(1)()), 2-(bis(2-pyridinylmethyl)amino)ethyl-beta-D xylopyranoside (L(2)()), and 2-(bis(2-pyridinylmethyl)amino)ethyl-alpha-d mannopyranoside (L(3)). An ethylene spacer is used to separate the carbohydrate moiety and the dipicolylamine (DPA) function in all three ligands. The Re complexes [Re(L(1-3))(CO)(3)]Br were characterized by (1)H and (13)C 1D/2D NMR spectroscopies, which confirmed the pendant nature of the carbohydrate moieties in solution. NMR measurements also established the long-range asymmetric effect of the carbohydrate functions on the chelating portion of the ligand. One analogue, [Re(L(1))(CO)(3)]Cl, was characterized in the solid state by X-ray crystallography. Further characterization was provided by IR spectroscopy, elemental analysis, conductivity, and mass spectrometry. Radiolabeling of L(1) L(3) with [(99m)Tc(H(2)O)(3)(CO)(3)](+) afforded high yield compounds of identical character to the Re analogues. The radiolabeled compounds were found to be stable toward ligand exchange in the presence of a large excess of either cysteine or histidine over a 24-h period. PMID- 15819556 TI - Release of iron from ferritin by aceto- and benzohydroxamic acids. AB - The release of iron from ferritin by aceto- and benzohydroxamic acids was studied at two different iron chelator concentrations (100 and 10 mM), at two pH values (7.4 and 5.2), and in the presence or absence of urea. Collectively, the results demonstrate that both aceto- and benzohydroxamic acids remove iron from ferritin. Aceto- and benzohydroxamic acids penetrate the ferritin shell and react directly with the iron core of the ferritin cavity probably forming mono(hydroxamate) iron(III) complexes which exit ferritin and react with the excess hydroxamate in the solution to produce bis(hydroxamate) iron(III) complexes. The sizes of both the benzohydroxamic acid and the mono(benzohydroxamate) iron(III) complex, 6 and 7 A, respectively, are larger than that of the ferritin channels which indicates the flexibility of the channels to allow the entry and exit of these molecules. The size of the hydroxamic acid influenced the effectiveness of the iron release from ferritin following the expected trend with smaller iron chelators showing greater effectiveness. Likewise, the percentage of iron removed from ferritin was pH-dependent; the percentage of iron removed at pH 5.2 was greater than that at pH 7.4. Finally, the presence of urea, capable of opening the ferritin channels, dramatically increased the effectiveness of the iron chelator in removing iron from ferritin, especially at pH 7.4. PMID- 15819557 TI - Bis(cyclopentadienyl)titanium complexes of naphthalene-1,8-dithiolates, biphenyl 2,2'-dithiolates, and related ligands. AB - Titanocene 1,8-dithiolato-naphthalene and titanocene 2,2'-dithiolato biphenyl are produced by the reaction of naphtho[1,8-cd]-1,2-dithiole [or the biphenyl] with titanocene dicarbonyl (Ti(II)) in toluene at room temperature. The pro-ligands 2,7-di(tert-butyl)naphtho[1,8-cd]-1,2-dithiole, 5,6-dihydroacenaphtho[5,6-cd]-1,2 dithiole, 4,5-dithioacephenanthrylene, and 13,14-dithiapicene have been used in similar reactions with titanocene dicarbonyl to investigate the effect of steric bulk and of varying the naphthalene backbone on the final complex. The resulting Cp(2)TiS(2)Ar complexes (Ar = naphthalene) have been shown by temperature dependent (1)H NMR spectroscopy to exist in solution in an envelope conformation with the six-membered TiS(2)C(3) rings undergoing inversion on the NMR time scale while the similar Cp(2)TiS(2)Ar complexes (Ar = biphenyl, binaphthalene) interconvert more rapidly. Titanocene 2,2'-disulfinato biphenyl has been synthesized by the salt elimination reaction of titanocene dichloride (Ti(IV)) and the disodium salt of biphenyl 2,2'-disulfinic acid. Finally, the effect of using pro-ligands where the sulfur atoms have been mono- or di-oxidized has been studied, and an interesting oxygen elimination reaction is observed for the S=O fragments but not for the SO(2) groups. All complexes have been characterized spectroscopically and seven X-ray structures are reported. PMID- 15819558 TI - Five new zinc phosphite structures: tertiary building blocks in the construction of hybrid materials. AB - The syntheses and structures of five new zinc phosphites [Zn(HPO(3))(C(4)H(6)N(2))] (1), [Zn(2)(HPO(3))(2)(C(10)H(10)N(2))(2)](2) (2), [Zn(HPO(3))(C(14)H(14)N(4))(0.5)] (3), [Zn(2)(HPO(3))(2)(C(14)H(14)N(4))].0.4H(2)O (4), and [Zn(2)(HPO(3))(2)(C(14)H(14)N(4))] (5) are reported. In compounds 1-3, the zinc atoms are ligated by 1-methylimidazole, 1-benzylimidazole, and 1,4-bis(imidazol-1 ylmethyl)benzene, respectively, while compounds 4 and 5 are synthesized in the presence of the same bifunctional ligand, 1,3-bis(imidazol-1-ylmethyl)benzene. The inorganic framework of compound 1 is composed of vertex-shared ZnO(3)N and HPO(3) tetrahedra that form 4-rings, which, in turn, are linked to generate a one dimensional ladder structure. In 2, the inorganic framework is composed of 4 rings and 8-rings to form the well-known 4.8(2) 2D network. This is connected via C-H...pi interactions between 1-benzylimidazole ligand to generate a pseudo pillared-layer structure. In 3, the inorganic framework again has the 4.8(2) topology pillared by the bis(imidazole) ligand, 1,3-bis(imidazol-1 ylmethyl)benzene. In 4, a new layer pattern is observed. Specifically, three edge sharing 4-rings form triple-fused 4-rings. These tertiary building units are further connected to form 12-rings. The alternating triple 4-rings and 12-rings form a previously unknown 2D inorganic sheet. The sheets are joined together by the bis(imidazole) ligand, 1,3-bis(imidazol-1-ylmethyl)benzene, to generate a 3D pillared-layer structure. In 4, benzene rings and imidazole rings stack in a zigzag pattern in the interlayer space. A significant role for the triple 4-ring tertiary building unit in the formation of hybrid inorganic/organic metal phosphite structures is proposed for 4 and 5. In 5, the triple 4-rings fuse to give a 1D stair-step structure. Calculations show that the triple 4-ring pattern observed in the linear ladder structure of 1 is more stable than that in the stair step pattern of 5. PMID- 15819560 TI - Isophthalato-based 2D coordination polymers of Eu(III), Gd(III), and Tb(III): enhancement of the terbium-centered luminescence through thiophene derivatization. AB - We present here the first examples of lanthanide ion complexes with only isophthalic acid or thiophenylisophthalic acid ligands. The complexes of isophthalic acid with Eu(3+) (1) and Tb(3+) (2) and the moderately soluble complexes of 5-thiophen-3-ylisophthalate with Eu(3+) (3), Gd(3+) (4), and Tb(3+) (5) were isolated as single crystals through gel crystallization. X-ray diffraction studies confirm the cross-linking structure of these complexes, which, in case of the thiophenyl derivatives, results in low solubility in common solvents. The two-dimensional isophthalato complex of Eu(3+) (1) crystallizes in the C2/c space group, with a = 22.154(4), b = 12.649(3), and c = 15.921(3) A, beta = 112.34(3) degrees, and V = 4126.7(14) A(3), while the one-dimensional Tb(3+) complex of the same ligand, 2, crystallizes in the space group P2(1)/c with a = 11.921(2), b = 10.838(2), and c = 17.499(4) A, beta = 92.44(3) degrees, and V = 2258.9(8) A(3). The thiophenylisophthalato complexes of Eu(3+) (3) and Gd(3+) (4) are two-dimensional and crystallize in the P2/n space group with parameters for 3 of a = 14.139(3), b = 10.684(2), and c = 15.138(3) A, beta = 102.51(3) degrees, and V = 2232.3(8) A(3) and parameters for 4 of a = 14.1195(13), b = 10.6594(10), and c = 15.1149(14) A, beta = 102.529(2) degrees, and V = 2220.7(4) A(3), while the Tb(3+) complex, 5, also two-dimensional, crystallizes in the P space group with a = 11.051(2), b = 14.528(3), and c = 15.041(3) A, alpha = 77.63(3), beta = 87.86(3), and gamma = 83.51(3) degrees, and V = 2343.48 A(3). All complexes of Eu(3+) and Tb(3+) luminesce in aqueous solution, and the luminescence lifetimes and quantum yields are 123.8 +/- 7.4, 0.14% (1), 475.1 +/- 14.5, 3.58% (3), 129.3 +/- 3.5, 0.19% (4), and 213.9 +/- 2.2 micros, 7.46% (5). PMID- 15819559 TI - A sugar's choice: coordination to a mononuclear or a dinuclear copper(II) complex? AB - We proposed a decisive role of the number of metal ions at the sugar binding site for carbohydrate-coordinating copper(II) complexes. To verify this hypothesis, we studied the binding of the representatively chosen carbohydrates D-ribose (7), D mannose (8), D-glucose (9), and D-maltose (10) to structurally related mono- and dinuclear copper(II) complexes in alkaline solution. All carbohydrates coordinate to the metal complexes in a 1:1 molar ratio. Coordination of 7 or 8 to the dinuclear copper(II) complex 1 is about 0.5 order of magnitude stronger than the complex formation with related mononuclear complexes. On contrast, 9, which is an epimer of 8, coordinates stronger to either one of the mononuclear copper(II) complexes in alkaline aqueous solution. PMID- 15819561 TI - Wheel-shaped [Mn12] single-molecule magnets. AB - The reaction of [Mn(12)O(12)(O(2)CCH(3))(16)(H(2)O)(4)].4H(2)O.2CH(3)COOH with n methyldiethanol amine (H(2)mdea), n-ethyldiethanol amine (H(2)edea), or n butyldiethanol amine (H(2)bdea) leads to the formation of wheel-shaped Mn(III)(6)Mn(II)(6) complexes with the general formula [Mn(12)(R)(O(2)CCH(3))(14)] (1, R = mdea; 2, R = edea; and 3, R = bdea). Complex 1 crystallizes in the triclinic space group P1, whereas complex 3 crystallizes in the monoclinic space group C(2/c). Complex 1a has the same molecular structure as complex 1 but crystallizes in the monoclinic space group P2(1/n). Complex 3a has the same molecular structure as complex 3 but crystallizes in the triclinic space group P1. Variable-temperature magnetic susceptibility data collected for complexes 1, 2, and 3 indicate that antiferromagnetic exchange interactions are present. The spin ground states of complexes 1, 2, and 3 were determined by fitting variable-field magnetization data collected in the 2-5 K temperature range. Fitting of these data yielded the spin ground-state parameters of S = 8, g = 2.0, and D = -0.47 cm(-1) for complex 1; S = 8, g = 2.0, and D = -0.49 cm(-1) for complex 2; and S = 8, g = 2, and D = -0.37 cm(-1) for complex 3. The ac magnetic susceptibility data were measured for complexes 1, 2, and 3 at temperatures between 1.8 and 10 K with a 3 G ac field oscillating in the range 50 1000 Hz. Slow kinetics of magnetization reversal relative to the frequency of the oscillating ac field were observed as frequency-dependent out-of-phase peaks for complexes 1, 2, and 3, and it can be concluded that these three complexes are single-molecule magnets. PMID- 15819562 TI - Temperature dependence of the crystal structure and g-values of [(HC(Ph2PO)3)2Cu](ClO4)2.2H2O: influence of dynamic Jahn-Teller coupling and lattice strain interactions. AB - The temperature dependence of the X-ray crystal structure and powder EPR spectrum of [(HC(Ph(2)PO)(3))(2)Cu](ClO(4))(2).2H(2)O is reported, and the structure at room temperature confirms that reported previously. Below approximately 100 K, the data imply a geometry with near elongated tetragonal symmetry for the [(HC(Ph(2)PO)(3))(2)Cu](2+) complex, but on warming the two higher Cu-O bond lengths and g-values progressively converge, and by 340 K the bond lengths correspond to a compressed tetragonal geometry. The data may be interpreted satisfactorily assuming an equilibrium among the energy levels of a Cu-O(6) polyhedron subjected to Jahn-Teller vibronic coupling and a lattice strain. However, agreement with the experiment is obtained only if the orthorhombic component of the lattice strain decreases to a negligible value as the temperature approaches 340 K. PMID- 15819563 TI - Field-induced magnetic transitions in metal phosphonates with ladderlike chain structures: (NH3C6H4NH3)M2(hedpH)2.H2O [M = Fe, Co, Mn, Zn; hedp = C(CH3)(OH)(PO3)2]. AB - This paper reports the syntheses and characterization of four isomorphous compounds (NH(3)C(6)H(4)NH(3))M(2)(hedpH)(2).H(2)O [M = Fe (1), Co (2), Mn (3), Zn (4); hedp = C(CH(3))(OH)(PO(3))(2)]. Each contains two crystallographically different kinds of {M(2)(hedpH)(2)}(n) double chains, where the {M(2)(mu-O)(2)} dimer units are connected by O-P-O bridges. The double chains are connected through extensive hydrogen bonds, hence generating a three-dimensional supramolecular network. The temperature-dependent magnetic susceptibility measurements show dominant antiferromagnetic interactions in compounds 1-3, mediated through the mu-O and/or O-P-O bridges between the metal(II) centers. The magnetization measurements reveal that compounds 1-3 experience field-induced magnetic transitions at low temperatures. PMID- 15819564 TI - Transformation of mu4-phosphinidenes at an Ru5 center: isolation and structural characterization of hydroxyphosphinidene cluster acids, fluorophosphinidenes, and a novel mu5-phosphide. AB - Acid hydrolysis of [Ru(5)(CO)(15)(mu(4)-PN(i)Pr(2))] (2) or protonation of the anionic PO cluster [Ru(5)(CO)(15)(mu(4)-PO)](-) (3) affords the hydroxyphosphinidene complex [Ru(5)(CO)(15)(mu(4) POH)].1.[H(2)N(i)()Pr(2)][CF(3)SO(3)], which cocrystallizes with a hydrogen bonded ammonium triflate salt. Reaction of [Ru(5)(CO)(15)(mu(4)-PN(i)Pr(2))] (2) with bis(diphenylphosphino)methane (dppm) leads to [Ru(5)(CO)(13)(mu-dppm)(mu(4) PN(i)Pr(2))] (4). Acid hydrolysis of 4 leads to the dppm-substituted hydroxyphosphinidene [Ru(5)(CO)(13)(mu-dppm)(mu(4)-POH)] (5), which is analogous to 1, but unlike 1, can be readily isolated as the free hydroxyphosphinidene acid. Compound 5 can also be formed by reaction of 3 with dppm and acid. The cationic hydride cluster [Ru(5)(CO)(13)(mu-dppm)(mu(3)-H)(mu(4)-POH)][CF(3)SO(3)] (6) can be isolated from the same reaction if chromatography is not used. Compound 4 also reacts with HBF(4) to form the fluorophosphinidene cluster [Ru(5)(CO)(13)(mu-dppm)(mu(4)-PF)] (7), while reaction with HCl leads to the mu chloro, mu(5)-phosphide cluster [Ru(5)(CO)(13)(mu-dppm)(mu-Cl)(mu(5)-P)] (8). PMID- 15819565 TI - Synthesis and Characterization of Ge(II), Sn(II), and Pb(II) Monoamides with -NH2 Ligands. AB - The synthesis and characterization of the first divalent germanium, tin, and lead monoamide derivatives of the parent amide group -NH(2) are presented. They have the general formula (ArMNH(2))(2) (M = Ge, Ar = Ar'(C(6)H(3)-2,6-Pr(i)(2)) or Ar* (C(6)H(3)-2,6(C(6)H(2)-2,4,6-Pr(i)(3))); M = Sn, Ar = Ar*; M = Pb, Ar = Ar*). For germanium and tin, they were obtained by reacting the corresponding terphenyl halides of the group 14 elements with liquid ammonia in diethyl ether. The lead amide derivative (Ar*PbNH(2))(2) was synthesized by reaction of LiNH(2) with Ar*PbBr in diethyl ether. The compounds were characterized by IR and multinuclear NMR spectroscopies and by X-ray crystallography in the case of the (Ar'GeNH(2))(2) or (Ar*SnNH(2))(2) derivatives. They possess dimeric structures with two -NH(2) groups bridging the germanium and tin centers. For lead, the reaction with ammonia led to isolation of a stable ammine complex of formula Ar*PbBr(NH(3)) which was characterized by IR and NMR spectroscopies and by X-ray crystallography. It is the first structural characterization of a divalent lead ammine complex. PMID- 15819566 TI - Release of NO from reduced nitroprusside ion. Iron-dinitrosyl formation and NO disproportionation reactions. AB - The kinetics and mechanism of the thermal decomposition of the one-electron reduction product of [Fe(CN)(5)NO](2-) (nitroprusside ion, NP) have been studied by using UV-vis, IR, and EPR spectroscopy and mass-spectrometric and electrochemical techniques in the pH range of 4-10. The reduction product contains an equilibrium mixture of [Fe(CN)(4)NO](2-) and [Fe(CN)(5)NO](3-) ions. The first predominates at pH <8 and is formed by the rapid release of trans cyanide from [Fe(CN)(5)NO](3-), which, in turn, is the main component at pH >9 10. Both nitrosyl complexes decay by first-order processes with rate constants around 10(-5) s(-1) (pH 6-10) related to the dissociation of NO. The decomposition is enhanced at pH 4 by 2 orders of magnitude with protons (and also metal ions) favoring the release of cyanides from the [Fe(CN)(4)NO](2-) ions and the ensuing rapid delivery of NO. At pH 7, an EPR-silent intermediate I(1) is detected (nu(NO), 1695 and 1740 cm(-1)) and assigned to the trans [Fe(II)(CN)(4)(NO)(2)](2-) ion, an {Fe(NO)(2)}(8) species. At pH 6-8, I(1) induces a disproportionation process with formation of N(2)O and the regeneration of nitroprusside in a 1:2 molar ratio. At lower pHs, I(1) leads, competitively, to a second paramagnetic (S = 1/2) dinitrosyl intermediate I(2), [Fe(CN)(2)(NO)(2)](1-), a new member of a series of four-coordinate {Fe(L)(2)(NO)(2)} complexes (L = thiolates, imidazole, etc.), described as {Fe(NO)(2)}(9). Other decomposition products are hexacyanoferrate(II) or free cyanide, depending on the pH, and precipitates of the Prussian-Blue type. This study throws light on the conditions favoring rapid release of NO, to promote vasodilatory effects upon NP injection, and describes new processes related to dinitrosyl formation and NO disproportionation, which are also relevant to the diverse biological processes associated with NO and N(2)O processing. PMID- 15819568 TI - Synthesis and structures of cis- and trans-bis(alkyneselenolato)platinum(II) complexes. AB - The reaction of 2 equiv of LiSeCC-n-C(5)H(11) (1) with cis-PtCl(2)(Ph(3)P)(2) (2) gives a mixture of the cis and trans isomers of Pt(Ph(3)P)(2)(SeCC-n C(5)H(11))(2) (3), which slowly isomerizes in CH(2)Cl(2) to the preferred trans form trans-3. The closely related cis-[Pt(dppf)(2)(SeCC-n-C(5)H(11))(2)] (4) (dppf = bis(diphenylphosphino)ferrocene) was prepared by a similar metathetical reaction using the platinum chloride complex of the chelating dppf to impose the cis geometry. The structures of the cis and trans complexes have been investigated in solution by heteronuclear NMR ((31)P, (77)Se, and (195)Pt) and, in the cases of trans-3 and 4, characterized in the solid state by single-crystal X-ray diffraction. Changing the coordination geometry from cis to trans induces significant changes in the structural and spectroscopic parameters, which do not comply with the previously anticipated donor-acceptor properties of selenolate ligands. PMID- 15819567 TI - Lewis acid properties of zinc(II) in Its cyclen complex. The structure of [Zn(cyclen)(S=C(NH2)2](ClO4)2 and the bonding of thiourea to metal ions. Some implications for zinc metalloenzymes. AB - The structure of the complex [Zn(cyclen)Tu](NO(3))(2) (1) is reported (cyclen = 1,4,7,10-tetraazacyclododecane; Tu = thiourea): orthorhombic, space group P2(1)2(1)2(1), a = 11.4170(11) A, b = 12.1995(11) A, c = 12.5299(12) A, Z = 4, R = 0.0504. The coordination of the cyclen is the same as that found for other similar Zn(II) complexes, with square pyramidal coordination around the Zn(II) and mean Zn-N bond lengths of 2.16 A. The coordinated Tu occupies the axial coordination site, with Zn-S = 2.31 A. The Zn-S-C-N torsion angle, involving the coordinated Tu, of 75.4 degrees is unusually large, because such torsion angles involving coordinated Tu are normally closer to 0 degrees. The bonding between Zn and S is discussed in terms of overlap with the p orbitals on S, which favors the eclipsed (Zn-S-C-N torsion = 0 degrees) mode of coordination of Tu. The energies of eclipsed and staggered modes (Zn-S-C-N = 90 degrees) of coordination of Tu to metal ions are examined by means of ab initio calculations, using the STO-3G basis set. It is concluded that the rather low formation constant for the Tu complex with Zn(II)/cyclen reported in this work was due to steric effects in 1, which prevent the adoption of the lower energy eclipsed conformation. These steric effects, because of clashes that would occur between Tu in the eclipsed conformation and the cyclen ring, cause the coordination of Tu with a higher energy conformation, with Zn-S-C-N = 75.4 degrees. The latter approaches the high energy staggered conformation that has Zn-S-C-N = 90 degrees. log K(1) values for Cl(-), Br(-), I(-), and CN(-) are reported and shown to be consistent with the binding site on the Zn(II) in the Zn(II)/cyclen complex being softer in the hard and soft acids and bases (HSAB, Pearson 1997) sense than the Zn(II) aqua ion, but not as soft as Zn(II) in triaza macrocycles that promote tetrahedral coordination. The change in HSAB character from intermediate in the Zn(II) aqua ion to softer in the cyclen complex, and softer still in tridentate N-donor ligands in model complexes, and in the Zn(II) active site of carbonic anhydrase as representative of Zn(II) metalloenzymes in general, is discussed in terms of the role of such effects in the functioning of metalloenzymes. PMID- 15819569 TI - Six-coordinate titanium complexes of a tripodal aminetris(phenoxide) ligand: synthesis, structure, and dynamics. AB - The five-coordinate titanium(IV) alkoxide LTi(O(t)Bu) (LH(3) = tris(2-hydroxy-3,5 di-tert-butylbenzyl)amine) is protonolyzed readily by the conjugate acids of monoanionic bidentate ligands, both symmetrical (tropolone, acetylacetone, di-p toluoylmethane) and unsymmetrical (8-hydroxyquinoline, salicylaldehyde, 2,6 diformyl-p-cresol, anthrarufin). The geometry of these complexes, which is pseudo octahedral with the tripodal ligand adopting a chiral, propeller-like conformation, has been confirmed in four cases by X-ray crystallography. Variable temperature NMR spectroscopy indicates that the six-coordinate complexes undergo two dynamic processes. First, the ligands undergo a twisting motion that results in racemization, a process which is over 10(4) times faster than in five coordinate complexes. The rate acceleration upon binding of an equatorial ligand is ascribed to steric repulsions with one of the cis phenoxides; the dynamics of a binuclear dibenzyl phosphate-bridged compound, which has a unique conformation of the tripodal ligand, indicates that flexing the cis phenoxide is the rate limiting step in racemization. Second, the complexes undergo a process that interchanges the inequivalent arms of the tripodal ligand. This process involves a trigonal twist that shifts the bidentate ligand between clefts in the tripod. The intermediate geometry in the reaction appears to be a transition state and not a long-lived intermediate, as judged from the relative rates of interconversion of tripod arms and chelate ends in the ditoluoylmethane complex. Tripod arm interchange takes place without partial dissociation of the bidentate chelate, a reaction that has been observed on a slower time scale in one case. PMID- 15819570 TI - Reactions of the dianion [1,1,1-(CO)3-2-Ph-closo-1,2-MnCB9H9]2- with transition metal cations: facile insertion and then extrusion of cluster vertexes. AB - The manganacarborane dianion in [N(PPh(3))(2)][NEt(4)][1,1,1-(CO)(3)-2-Ph-closo 1,2-MnCB(9)H(9)] (1b) reacts with cationic transition metal-ligand fragments to give products in which the electrophilic metal groups (M') are exo-polyhedrally attached to the {closo-1,2-MnCB(9)} cage system via three-center two-electron B-H --> M' linkages and generally also by Mn-M' bonds. With {Cu(PPh(3))}(+), the Cu Mn-Cu trimetallic species [1,6-{Cu(PPh(3))}-1,7-{Cu(PPh(3))}-6,7-(mu-H)(2)-1,1,1 (CO)(3)-2-Ph-closo-1,2-MnCB(9)H(7)] (3a) is formed, whereas reactions with {M'(dppe)}(2+) (M' = Ni, Pd; dppe = Ph(2)PCH(2)CH(2)PPh(2)) give [1,3-{Ni(dppe)} 3-(mu-H)-1,1,1-(CO)(3)-2-Ph-closo-1,2-MnCB(9)H(8)] (5a) and [1,3,6-{Pd(dppe)}-3,6 (mu-H)(2)-1,1,1-(CO)(3)-2-Ph-closo-1,2-MnCB(9)H(7)] (5b), both of which contain M'-Mn bonds. The latter reaction with M' = Pt affords [3,6-{Pt(dppe)}-3,6-(mu H)(2)-1,1,1-(CO)(3)-2-Ph-closo-1,2-MnCB(9)H(7)] (6), which lacks a Pt-Mn connectivity. Compound 6 itself spontaneously converts to [1-Ph-2,2,2-(CO)(3)-8,8 (dppe)-hypercloso-8,2,1-PtMnCB(9)H(9)] (7b) and thence to [3,6,7-{Mn(CO)(3)}-3,7 (mu-H)(2)-1-Ph-6,6-(dppe)-closo-6,1-PtCB(8)H(6)] (8). This sequence occurs via initial insertion of the {Pt(dppe)} unit and then extrusion of {Mn(CO)(3)} and one {BH} vertex. In the presence of alcohols ROH, compound 6 is transformed to the 7-OR substituted analogues of 7b. X-ray diffraction studies were essential in elucidating the structures encountered in compounds 5-8 and hence in understanding their behavior. PMID- 15819571 TI - Synthesis, characterization, and photochemical behavior of {Ru(arene)}2+ derivatives of alpha-[PW11O39]7-: an organometallic way to ruthenium-substituted heteropolytungstates. AB - Reaction of [Ru(arene)Cl(2)](2) (arene = benzene, toluene, p-cymene, hexamethylbenzene) with K(7)[PW(11)O(39)].14H(2)O provided two series of organometallic derivatives of heteropolytungstates: type-1 and type-2 complexes of general formulas [PW(11)O(39){Ru(arene)(H(2)O)}](5-) and [{PW(11)O(39){Ru(arene)}}(2){WO(2)}](8-), respectively. All compounds were characterized by infrared and multinuclear NMR ((1)H, (31)P, (183)W) spectroscopies. The crystal structures of Na(4)K(4)[{PW(11)O(39){Ru(benzene)}}(2){WO(2)}].6H(2)O (NaK-2a.6H(2)O), K(7)H[{PW(11)O(39){Ru(toluene)}}(2){WO(2)}].4H(2)O (K-2b.4H(2)O), and Cs(3)K(2)[PW(11)O(39){Ru(p-cymene)(H(2)O)}].4H(2)O (CsK-1c.4H(2)O) were obtained and revealed that the {Ru(arene)} fragment is supported on the oxometallic framework. Photochemical reactivity of [PW(11)O(39){Ru(arene)(H(2)O)}](5-) (arene = toluene, p-cymene) in the presence of various ligands L (L = H(2)O, dimethyl sulfoxide, tetramethylene sulfoxide, and diphenyl sulfoxide) was investigated, and led to the formation of [PW(11)O(39){Ru(L)}](5-), in which the ruthenium is incorporated into the lacunary [PW(11)O(39)](7-) anion. PMID- 15819572 TI - Synthesis, characterization, and photophysical properties of a free base and a biszinc(II) complex of 1,3-bisporphyrincalix[4]arene: evidence for "tunable intramolecular open and closed conformations". AB - The bismacrocycle 5,17-bis[5-(2,8,13,17-tetraethyl-3,7,12,18 tetramethylporphyrinyl)]-25,26,27,28-tetrapropoxycalix[4]arene (4) was synthesized in three steps from the corresponding bisaldehyde 5,17-diformyl 25,26,27,28-tetrapropoxycalix[4]arene. The biszinc(II) complex (5) was prepared as well, and the photophysical properties were measured using 2-MeTHF as solvent at 298 and 77 K. While computer modeling for 5 predicts that both pinched cone conformers, closed (porphyrins near each other) and open (porphyrins away from each other), may exist in the "gas phase", the experimental data indicate clearly that no zinc porphyrin...zinc porphyrin interactions are present in solution at 298 K, favoring the open conformer, where the two macrocycles are placed away from each other. On the other hand, clear evidence for a closed conformer is observed at 77 K. Variable-temperature (1)H NMR experiments show that 5 is fluxional between 298 and 183 K, while 4 keeps the open conformation for the whole temperature range. This behavior is unprecedented for calix[4]areneporphyrin compounds, and the relative porphyrin ring rigidity is postulated to explain this difference. PMID- 15819573 TI - The redox series [M(bpy)2(q)]n+, M = Ru or Os, Q = 3,5-di-tert-butyl-n-phenyl-1,2 benzoquinonemonoimine. Isolation and a complete X and W band EPR study of the semiquinone states (n = 1). AB - The complexes [M(bpy)(2)(Q)](PF(6)) (bpy = 2,2'-bipyridyl; M = Ru, Os; Q = 3,5-di tert-butyl-N-phenyl-1,2-benzoquinonemonoimine) were isolated and studied by X and W band EPR in a dichloromethane solution at ambient temperatures and at 4 K. For M = Ru, the (14)N hyperfine splitting confirms the Ru(II)/semiquinone formulation, although at a > 1 mT, the (99,101)Ru satellite coupling is unusually high. W band EPR allowed us to determine the relatively small g anisotropy Delta g = g(1) - g(3) = 0.0665 for the ruthenium complex. The osmium analogue exhibits a much higher difference Delta g = 0.370, which is attributed not only to the larger spin-orbit coupling constant of Os versus that of Ru but also to a higher extent of metal contribution to the singly occupied molecular orbital. The difference Delta E between the oxidation and reduction potentials of the radical complexes is larger for the ruthenium compound (Delta E = 0.87 V) than for the osmium analogue (Delta E = 0.72), confirming the difference in metal/ligand interaction. The electrochemically generated states [M(bpy)(2)(Q)](n+), n = 0, 1, 2, and 3, were also characterized using UV-vis-near-infrared spectroelectrochemistry. PMID- 15819574 TI - Magnetic resonance spectroscopic investigations of the electronic ground and excited states in strongly nonplanar iron(III) dodecasubstituted porphyrins. AB - A series of axially ligated complexes of iron(III) octamethyltetraphenylporphyrin, (OMTPP)Fe(III), octaethyltetraphenylporphyrin, (OETPP)Fe(III), its perfluorinated phenyl analogue, (F(20)OETPP)Fe(III), and tetra-(beta,beta'-tetramethylene)tetraphenylporphyrin, (TC(6)TPP)Fe(III), have been prepared and characterized by (1)H NMR spectroscopy: chloride, perchlorate, bis-4-(dimethylamino)pyridine, bis-1-methylimidazole, and bis-cyanide. Complete spectral assignments have been made using 1D and 2D techniques. The temperature dependences of the proton resonances of the complexes show significant deviations from simple Curie behavior and evidence of ligand exchange, ligand rotation, and porphyrin ring inversion at ambient temperatures. At temperatures below the point where dynamics effects contribute, the temperature dependences of the proton chemical shifts of the complexes could be fit to an expanded version of the Curie law using a temperature-dependent fitting program developed in our laboratory that includes consideration of a thermally accessible excited state. The results show that, although the ground state differs for various axial ligand complexes and is usually fully consistent with that observed by EPR spectroscopy at 4.2 K, the excited state often has S = (3)/(2) (or S = (5)/(2) in the cases where the ground state has S = (3)/(2)). The EPR spectra (4.2 K) of bis-4 (dimethylamino)pyridine and bis-1-methylimidazole complexes show "large-g(max)" signals with g(max) = 3.20 and 3.12, respectively, and the latter also shows a normal rhombic EPR signal, indicating the presence of low-spin (LS) (d(xy))(2)(d(xz),d(yz))(3) ground states for both. The bis-cyanide complex also yields a large-g(max) EPR spectrum with g = 3.49 and other features that could suggest that some molecules have the (d(xz),d(yz))(4)(d(xy))(1) ground state. The EPR spectra of all five-coordinate chloride complexes have characteristic features of predominantly S = (5)/(2) ground-state systems with admixture of 1 10% of S = (3)/(2) character. PMID- 15819575 TI - Kinetics of ring inversion in strongly nonplanar iron(III) octaalkyltetraphenylporphyrinates. AB - The dynamics of porphyrin ring inversion of a number of Fe(III) complexes of octamethyltetraphenylporphyrin, (OMTPP)Fe(III); octaethyltetraphenylporphyrin, (OETPP)Fe(III); octaethyltetra(perfluorophenyl)porphyrin, (F(20)OETPP)Fe(III); and tetra-beta,beta'-tetramethylenetetraphenyl-porphyrin, (TC(6)TPP)Fe(III), having either one (Cl(-), ClO(4-)) or two [4-(dimethylamino)pyridine, 4-Me(2)NPy; 1-methylimidazole, 1-MeIm; tert-butylisocyanide, t-BuNC; or cyanide, CN(-)] axial ligands have been characterized by 1D dynamic NMR (DNMR) and 2D (1)H NOESY/EXSY spectroscopies as a function of temperature. The activation parameters, Delta H++, Delta S++, and Delta G++(298), and the extrapolated rate constants at 298 K for three chloride, one perchlorate, and three bis-(4-Me(2)NPy) complexes as well as [FeOETPP(1-MeIm)(2)]Cl, [FeOETPP(t-BuNC)(2)]ClO(4), and Na[FeOETPP(CN)(2)] have been determined. The results indicate that there is a wide range of flexibility for the porphyrin core (k(ex)(298) = 10-10(7) s(-1)) that decreases in the order TC(6)TPP > OMTPP > F(20)OETPP > or = OETPP, which correlates with increasing porphyrin nonplanarity. To determine the effect of axial ligands, we calculated the free energy of activation, Delta G++(298) for OETPPFe(III) bis ligated with 4-Me(2)NPy, 1-MeIm, or 4-CNPy (approximately 59 kJ mol(-1)), and for complexes with small cylindrical ligands (t-BuNC and CN(-)) (approximately 37 kJ mol(-1)). These data suggest that the Delta G++(298) for planar ligand rotation is roughly 20-25 kJ mol(-1). PMID- 15819576 TI - New titanium imido synthons: syntheses and supramolecular structures. AB - Reactions of Ti(NMe(2))(2)Cl(2) with a wide range of primary alkyl and arylamines RNH(2) afforded the corresponding 5-coordinate imido titanium compounds Ti(NR)Cl(2)(NHMe(2))(2) (R = (t)Bu (1), (i)Pr (2), CH(2)Ph (3), Ph (4), 2,6 C(6)H(3)Me(2) (5), 2,6-C(6)H(3)(i)Pr(2) (6), 2,4,6-C(6)H(2)F(3) (7), 2,3,5,6 C(6)HF(4) (8), C(6)F(5) (9), 4-C(6)H(4)Cl (10), 2,3,5,6-C(6)HCl(4) (11), 2 C(6)H(4)CF(3) (12), 2-C(6)H(4)(t)Bu (13)). The compounds 1-13 are monomeric in solution but in the solid state form either N-H...Cl hydrogen bonded dimers or chains or perfluorophenyl pi-stacked chains, depending on the imido R-group. The compound 13 was also prepared in a "one-pot" synthesis from RNH(2) and Ti(NMe(2))(4) and Me(3)SiCl. Reaction of certain Ti(NR)Cl(2)(NHMe(2))(2) compounds with an excess of pyridine afforded the corresponding bis- or tris pyridine analogues [Ti(NR)Cl(2)(py)(x)](y) (x = 3, y = 1; x = y = 2), and the structure of Ti(2)(NC(6)F(5))(2)Cl(2)(mu-Cl)(2)(py)(4) shows pi-stacking of perfluorophenyl rings. Reaction of Ti(NMe(2))(2)Cl(2) with cross-linked aminomethyl polystyrene gave quantitative conversion to the corresponding solid supported titanium imido complex. This paper represents the first detailed study of how supramolecular structures of imido compounds may be influenced by simple variation of the imido ligand N-substituent. PMID- 15819577 TI - Reactions of (PCP)Ru(CO)(NHPh)(PMe3) (PCP = 2,6-(CH2PtBu2)2C6H3) with substrates that possess polar bonds. AB - The Ru(II) amido complex (PCP)Ru(CO)(PMe(3))(NHPh) (1) (PCP = 2,6 (CH(2)P(t)Bu(2))(2)C(6)H(3)) reacts with compounds that possess polar C=N, C triple bond N, or C=O bonds (e.g., nitriles, carbodiimides, or isocyanates) to produce four-membered heterometallacycles that result from nucleophilic addition of the amido nitrogen to an unsaturated carbon of the organic substrate. Based on studies of the reaction of complex 1 with acetonitrile, the transformations are suggested to proceed by dissociation of trimethylphosphine, followed by coordination of the organic substrate and then intramolecular N-C bond formation. In the presence of ROH (R = H or Me), the fluorinated amidinate complex (PCP)Ru(CO)(N(Ph)C(C(6)F(5))NH) (6) reacts with excess pentafluorobenzonitrile to produce (PCP)Ru(CO)(F)(N(H)C(C(6)F(5))NHPh) (7). The reaction with MeOH also produces o-MeOC(6)F(4)CN (>90%) and p-MeOC(6)F(4)CN (<10%). Details of the solid state structures of (PCP)Ru(CO)(F)(N(H)C(C(6)F(5))NHPh) (7), (PCP)Ru(CO)[PhNC{NH(hx)}N(hx)] (8), (PCP)Ru(CO){N(Ph)C(NHPh)O} (9), and (PCP)Ru(CO){OC(Ph)N(Ph)} (10) are reported. PMID- 15819578 TI - Conformations, protonation sites, and metal complexation of benzohydroxamic Acid. A theoretical and experimental study. AB - A theoretical and experimental study on the structure and deprotonation of benzohydroxamic acid (BHA) has been performed. Calculations at the RHF/cc-pVDZ level, refined by the B3LYP/AUG-cc-pVDZ method, indicate that, in the gas phase, Z amide is the most stable structure of both neutral and deprotonated BHA. (1)H (1)H nuclear Overhauser enhancement spectroscopy and (1)H-(1)H correlation spectroscopy spectra in acetone, interpreted with ab initio interatomic distances, reveal that BHA is split into the Z and E forms, the [E]/[Z] ratio being 75:25 at -80 degrees C. The formation of E-E, Z-Z, and E-Z dimers has been detected; in the presence of water, the dimers dissociate to the corresponding monomers. The rates of proton exchange within the Z and E forms and between E and Z were measured by dynamic (1)H NMR in the -60 to 40 degrees C temperature range; an increase in water content lowers the rate of exchange of the E isomer. The effect of D(2)O on the NMR signals indicates a fast hydrogen exchange between D(2)O and the E and Z amide forms. The sequence of the acid strength at low temperatures is (N)H(E)) approximately (O)H(E) < (O)H(Z) approximately (N)H(Z). The kinetics of complex formation between BHA and Ni(2+), investigated by the stopped-flow method, show that both neutral BHA and its anion can bind Ni(2+). Whereas the anion reacts at a "normal" speed, the rate of water replacement from Ni(H(2)O)(6)(2+) by neutral BHA is about 1 order of magnitude less than expected. This behavior was interpreted assuming that, in aqueous solution, BHA mainly adopts a closed (hydrogen-bonded) Z configuration, which should open (with an energy penalty) for the metal binding process to occur. PMID- 15819579 TI - A ladderlike chain aluminum fluoride ([Al2F8]2-)n with edge-sharing AlF6 octahedra. AB - A new aluminum fluoride, Al(2)F(8).2NC(5)H(6).C(6)H(3)(CO(2)H)(3), was synthesized under mild hydrothermal conditions (200 degrees C, 3 days) in the presence of 1,3,5-benzenetricarboxylic acid (btc) in pyridine/HF (pyr/HF) solvent. Its structure is characterized with single-crystal XRD analysis and high resolution solid-state NMR. The inorganic framework consists of the corner- and edge-shared connections of AlF(6) octahedra. They are linked via a common edge and form a bioctahedral motif which is trans-connected through the corner-shared fluorine. It results in the formation of an original infinite double file of AlF(6) octahedra running along the a axis. A high-power decoupled MAS (27)Al{(19)F} Hahn echo NMR spectrum allowed us to distinguish the two crystallographic hexacoordinated Al sites. Four unresolved (19)F NMR signals are observed in the MAS spectra to account for the eight crystallographic fluorine atoms. Half of the terminal fluorine sites interact via strong hydrogen bonds with the ammonium groups of the pyridine moieties. The resulting mixed pyridine fluoroaluminate chains are intercalated by the btc molecules which are hydrogen bonded to the remaining free terminal fluoride anions through the protonated carboxylic acid function. The (1)H nuclei of both organic molecules are observed in the protonated form. PMID- 15819580 TI - The insertion of carbodiimides into Al and Ga amido linkages. Guanidinates and mixed amido guanidinates of aluminum and gallium. AB - The insertion of carbodiimides into existing metal-heteroatom bonds is an important preparative route for the synthesis of useful ligand systems such as amidinates and guanidinates. Our interest lies in multiple insertions at one metal center and the mechanisms of insertion and rearrangement. We have synthesized and characterized [Me(2)NC(N(i)Pr)(2)](n)M(NMe(2))(3)(-)(n) (n = 1, 2, 3; M = Al, Ga). We have investigated the mechanism of synthesis and discovered a ligand transfer step that is important for the formation of the final products. PMID- 15819582 TI - Kinetic and mechanistic study of the Pt(II) versus Pt(IV) effect in the platinum mediated nitrile-hydroxylamine coupling. AB - The metal-mediated coupling between coordinated EtCN in the platinum(II) and platinum(IV) complexes cis- and trans-[PtCl(2)(EtCN)(2)], trans [PtCl(4)(EtCN)(2)], a mixture of cis/trans-[PtCl(4)(EtCN)(2)] or [Ph(3)PCH(2)Ph][PtCl(n)(EtCN)] (n = 3, 5), and dialkyl- and dibenzylhydroxylamines R(2)NOH (R = Me, Et, CH(2)Ph, CH(2)C(6)H(4)Cl-p) proceeds smoothly in CH(2)Cl(2) at 20-25 degrees C and the subsequent workup allowed the isolation of new imino species [PtCl(n){NH=C(Et)ONR(2)}(2)] (n = 2, R = Me, cis-1 and trans-1; Et, cis-2 and trans-2; CH(2)Ph, cis-3 and trans-3; CH(2)C(6)H(4)Cl p, cis-4 and trans-4; n = 4, R = Me, trans-9; Et, trans-10; CH(2)Ph, trans-11; CH(2)C(6)H(4)Cl-p, trans-12) or [Ph(3)PCH(2)Ph][PtCl(n){NH=C(Et)ONR(2)}] (n = 3, R = Me, 5; Et, 6; CH(2)Ph, 7; CH(2)C(6)H(4)Cl-p, 8; n = 5, R = Me, 13; Et, 14; CH(2)Ph, 15; CH(2)C(6)H(4)Cl-p, 16) in excellent to good (95-80%) isolated yields. The reduction of the Pt(IV) complexes 9-16 with the ylide Ph(3)P=CHCO(2)Me allows the synthesis of Pt(II) species 1-8. The compounds 1-16 were characterized by elemental analyses (C, H, N), FAB-MS, IR, (1)H, (13)C{(1)H}, and (31)P{(1)H} NMR (the latter for the anionic type complexes 5-8 and 13-16) and by X-ray crystallography for the Pt(II) (cis-1, cis-2, and trans 4) and Pt(IV) (15) species. Kinetic studies of addition of R(2)NOH (R = CH(2)C(6)H(4)Cl-p) to complexes [Ph(3)PCH(2)Ph][Pt(II)Cl(3)(EtCN)] and [Ph(3)PCH(2)Ph][Pt(IV)Cl(5)(EtCN)] by the (1)H NMR technique revealed that both reactions are first order in (p-ClC(6)H(4)CH(2))(2)NOH and Pt(II) or Pt(IV) complex, the second-order rate constant k(2) being three orders of magnitude larger for the Pt(IV) complex. The reactions are intermolecular in nature as proved by the independence of k(2) on the concentrations of added EtC triple bond N and Cl(-). These data and the calculated values of Delta H++ and Delta S++ are consistent with the mechanism involving the rate-limiting nucleophilic attack of the oxygen of (p-ClC(6)H(4)CH(2))(2)NOH at the sp-carbon of the C triple bond N bond followed by a fast proton migration. PMID- 15819581 TI - Chromium(V) complexes of hydroxamic acids: formation, structures, and reactivities. AB - A new family of relatively stable Cr(V) complexes, [Cr(V)O(L)(2)](-) (LH(2) = RC(O)NHOH, R = Me, Ph, 2-HO-Ph, or HONHC(O)(CH(2))(6)), has been obtained by the reactions of hydroxamic acids with Cr(VI) in polar aprotic solvents. Similar reactions in aqueous solutions led to the formation of transient Cr(V) species. All complexes have been characterized by electron paramagnetic resonance spectroscopy and electrospray mass spectrometry. A Cr(V) complex of benzohydroxamic acid (1, R = Ph) was isolated in a pure form (as a K(+) salt) and was characterized by X-ray absorption spectroscopy and analytical techniques. Multiple-scattering analysis of X-ray absorption fine structure spectroscopic data for 1 (solid, 10 K) point to a distorted trigonal-bipyramidal structure with trans-oriented Ph groups and Cr-ligand bond lengths of 1.58 A (Cr-O), 1.88 A (Cr O(C)), and 1.98 A (Cr-O(N)). Under ambient conditions, 1 is stable for days in aprotic solvents but decomposes within minutes in aqueous solutions (maximal stability at pH approximately 7), which leads predominantly to the formation of Cr(III) complexes. Complex 1 readily undergoes ligand-exchange reactions with biological 1,2-diols, including D-glucose and mucin, in neutral aqueous solutions. It differs from most other types of Cr(V) complexes in its biological activity, since no oxidative cleavage of plasmid DNA in vitro and no significant bacterial mutagenicity (in the TA 102 strain of Salmonella typhimurium) was observed for 1. In natural systems, stabilization of Cr(V) by hydroxamato ligands from bacterial-derived siderophores (followed by ligand-exchange reactions with more abundant carbohydrate ligands) may occur during the biological reduction of Cr(VI) in contaminated soils. PMID- 15819583 TI - Ground states, excited states, and metal-ligand bonding in rare earth hexachloro complexes: a DFT-based ligand field study. AB - Metal (4f)-ligand (Cl 3p) bonding in LnCl(6)(3-) (Ln = Ce to Yb) complexes has been studied on the basis of 4f-->4f and Cl,3p-->4f charge-transfer spectra and on the analysis of these spectra within the valence bond configuration interaction model to show that mixing of Cl 3p into the Ln 4f ligand field orbitals does not exceed 1%. Contrary to this, Kohn-Sham formalism of density functional theory using currently available approximations to the exchange correlation functional tends to strongly overestimate 4f-3p covalency, yielding, for YbCl(6)(3-), a much larger mixing of Cl 3p-->4f charge transfer into the f(13) ionic ground-state wave function. Thus, ligand field density functional theory, which was recently developed and applied with success to complexes of 3d metals in our group, yields anomalously large ligand field splittings for Ln, the discrepancy with experiment increasing from left to the right of the Ln 4f series. It is shown that eliminating artificial ligand-to-metal charge transfer in Kohn-Sham calculations by a procedure described in this work leads to energies of 4f-4f transitions in good agreement with experiment. We recall an earlier concept of Ballhausen and Dahl which describes ligand field in terms of a pseudopotential and give a thorough analysis of the contributions to the ligand field from electrostatics (crystal field) and exchange (Pauli) repulsion. The close relation of the present results with those obtained using the first principles based and electron density dependent effective embedding potential is pointed out along with implications for applications to other systems. PMID- 15819584 TI - Thermodynamics of the As(III)-thiol interaction: arsenite and monomethylarsenite complexes with glutathione, dihydrolipoic acid, and other thiol ligands. AB - Colorimetric (near-UV absorption spectroscopy) and calorimetric (isothermal titration calorimetry) methods have been used to quantify the equilibrium and thermodynamics of arsenite and monomethylarsenite (MMA) coordinating to glutathione (GSH) and the dithiols dimercaptosuccinic acid (DMSA), dihydrolipoic acid (DHLA), and dithiothreitol (DTT). We found that both arsenite and MMA form moderately stable complexes (beta = 10(6)-10(7)) with GSH; that arsenite forms a particularly stable 2:3 complex (beta approximately 10(18)) with the biological cofactor DHLA; that MMA has a somewhat higher affinity than arsenite for thiol ligands; and that entropic factors modulate the overall stability of As(III) complexes with thiols, which are favored by the exothermic formation of As(III) thiolate bonds. The implications of these results for arsenic toxicity are discussed. PMID- 15819585 TI - Formation of N-I charge-transfer bonds and ion pairs in polyiodides with imidotellurium cations. AB - [((t)BuNH)Te(mu-N(t)Bu)(2)Te(N(t))Bu)][OSO(2)CF(3)] (4a) is obtained in quantitative yields by the treatment of [((t)BuN)Te(mu-N(t)Bu)(2)Te(N(t)Bu)] (1) with HCF(3)SO(3). The reaction of 4a with LiI and iodine in the molar ratio 1:1:4.5 affords a product that, upon recrystallization from acetonitrile, was found to be a solid solution of [((t)BuNH)Te(mu-N(t)Bu)(2)Te(N(t)Bu)](2)I(20) (5a) and [((t)BuNH)Te(mu-N(t)Bu)(2)Te(NH(t)Bu)](2)I(18) (5b). Consequently, the crystal structure is disordered, containing 88.3(1)% of 5a.2MeCN and 11.7(1)% of 5b.2MeCN. The I(20) framework is involved in two symmetry-equivalent N-I-I-I-I fragments, two I(3)(-) ions, and three I(2) molecules that are linked together by I...I secondary bonding interactions. The bonding in the N-I-I-I-I fragment can be considered in terms of the lp(N) --> sigma*(I(2)) and pi(I(2)) --> sigma*(I(2)) charge-transfer interactions involving one [((t)BuNH)Te(mu N(t)Bu)(2)Te(N(t)Bu)](+) cation and two I(2) units. The N-I bond length of 2.131(7) A, the I-I distances of 3.118(1), 3.095(2), and 2.788(2) A, and the angle I(2)-I(2) angle of 84.75(4) degrees are consistent with this bonding scheme. The I-I bond distances in the two symmetry-equivalent I(3)(-) ions are 3.113(1) and 2.792(2) A, and those in two crystallographically independent I(2) molecules are 2.736(2) and 2.743(1) A. The formal I(18)(4)(-) anion in 5b.2MeCN consists of four I(3)(-) anions and three I(2) molecules linked by I...I secondary bonds. One crystallographically independent I(3)(-) anion is connected to the [((t)BuNH)Te(mu-N(t)Bu)(2)Te(HN(t)Bu)](2+) cation by two hydrogen bonds [H...I = 2.823(5) and 2.983(5) A; N...I = 3.697(8) and 3.857(9) A]. The I(3)(-) anions and I(2) molecules in 5b show virtually identical bond parameters to those in 5a. The treatment of 1 with iodine and the reactions of its methylated derivatives, [((t)BuNMe)Te(mu-N(t)Bu)(2)Te(N(t)()Bu)][OSO(2)CF(3)] and [((t)BuNMe)Te(mu-N(t)Bu)(2)Te(MeN(t)Bu)][OSO(2)CF(3)](2), with LiI and iodine also afford highly moisture-sensitive polyiodides, either by the formation of N-I charge-transfer complexes or by ionic interactions. The crystal structures of the partially hydrolyzed products, [((t)BuIN)Te(mu-N(t))Bu)(2)Te(mu-O)](2)(I(3))(2) (3), [((t)BuMeN)Te(mu-N(t)Bu)(2)Te(mu-O)](2)(I(3))(2) (6), and 6.2MeCN, are also reported. PMID- 15819586 TI - Optimal duration of antiplatelet therapy in recipients of coronary drug-eluting stents. AB - Four weeks' therapy with clopidogrel, in addition to aspirin (acetylsalicylic acid), is currently standard care after percutaneous coronary intervention (PCI) with stent implantation. The recent availability of drug-eluting stents (DES), which dramatically reduce restenosis at the site of PCI, has again raised the issue of stent thrombosis. In clinical trials, the risk of stent thrombosis appeared unrelated to the presence of the drug eluting from the stent and was documented within the usual range of < or =1% at 9 months after DES implantation. However, these devices feature delayed strut endothelialisation and there are reports describing late DES thrombosis up to 18 months after PCI, in most cases after clopidogrel has been discontinued. Although infrequent after bare-metal stenting (0.4-2.8%), stent thrombosis is a catastrophic event. Before DES availability, adjunctive intravascular brachytherapy significantly reduced in stent neointimal proliferation, at the price of a higher-than-expected rate of late stent occlusion (6-8%). In such setting, a 12-month aspirin plus clopidogrel regimen showed a beneficial effect on long-term adverse events. An additional consideration is that, among patients undergoing bare-metal stent PCI, combined antithrombotic therapy with aspirin and clopidogrel has been recently associated with favourable effects on cardiovascular outcome beyond stent thrombosis in two large-scale clinical trials. Therefore, we propose that prolonged combination therapy with aspirin and clopidogrel be mandatory up to 1 year after PCI in all patients receiving DES. PMID- 15819587 TI - Improving outcomes in advanced malignant melanoma: update on systemic therapy. AB - Malignant melanoma continues to increase in incidence throughout the developed world. Surgery remains the cornerstone of curative treatment and the use of adjuvant systemic therapy has provoked much debate. Metastatic disease is incurable in most patients. While combination chemotherapy or biochemotherapy may be considered in certain circumstances, it is now clear that single-agent chemotherapy remains the mainstay of treatment for the majority of patients.A number of new agents and novel approaches are under evaluation and show promise. The pro-apoptotic agent oblimersen has shown improved progression-free survival and response rate, although not overall survival, when combined with dacarbazine compared with dacarbazine alone. The BRaf inhibitor sorafenib (BAY 43-9006) has produced encouraging results when administered with chemotherapy and is now being assessed in randomised studies. Thalidomide in combination with chemotherapy is well tolerated and shows a trend towards increased clinical efficacy compared with chemotherapy alone. Other anti-angiogenic drugs, such as bevacizumab, are being investigated in trials. Results with tumour vaccines have been mixed and several large trials are ongoing. This paper discusses recent pivotal studies and promising new agents in systemic therapy for advanced malignant melanoma. PMID- 15819588 TI - Anaesthetic agents for advanced regional anaesthesia: a North American perspective. AB - Interest in the use of regional anaesthesia, particularly peripheral nerve blocks (PNBs) and continuous PNBs, has increased in recent years. Accompanying this resurgence in interest has been the development of new local anaesthetics and additives designed to enhance block duration and quality. This manuscript provides a literature-based review on accepted uses of local anaesthetics and adjuncts for a variety of regional anaesthesia techniques. A brief review of local anaesthetic pharmacodynamics describes the action of these drugs in preventing nerve depolarisation, thus blocking nerve impulses. Toxic adverse effects of local anaesthetics, specifically CNS and cardiac manifestations of excessive local anaesthetic blood concentrations and the direct neurotoxic properties of local anaesthetics, are discussed generally and specifically for many commonly used local anaesthetics. Clinically useful ester and amide local anaesthetics are evaluated individually in terms of their physical properties and toxic potential. How these properties impact on the clinical uses of each local anaesthetic is explored. Particular emphasis is placed on the long-acting local anaesthetic toxic potential of racemic bupivacaine compared with levobupivacaine and ropivacaine, which are both levorotatory stereoisomers. Guidelines for using ropivacaine and mepivacaine, based on the authors' experience using advanced regional anaesthesia in a busy practice, is provided. Finally, epinephrine (adrenaline), clonidine and other local anaesthetic additives and their rationale for use is covered along with other future possibilities. PMID- 15819589 TI - Bronchiolitis obliterans syndrome: risk factors and therapeutic strategies. AB - Post-transplantation bronchiolitis obliterans syndrome (BOS) is a clinicopathological syndrome characterised histologically by obliterative bronchiolitis (OB) and physiologically by airflow limitation. It affects long term survival with no consistently effective treatment strategy.An updated review of risk factors for OB/BOS, and approaches to prevention and treatment was performed through a systematic review of relevant studies between January 1990 and February 2005. The initial search identified 853 publications, with 56 articles reviewed after exclusions. Early acute rejection is the most significant risk factor, with late rejection (> or =3 months) also significant. Lymphocytic bronchitis/bronchiolitis is relevant, with later onset associated with greater risk. Viral infections are identified as significant risk factors. Human leukocyte antigen matching and OB/BOS development is a weaker association, but is stronger with acute rejection. Recipient and donor characteristics have a minor role. There is limited evidence that altering immunosuppression is effective in reducing the rate of decline in lung function. BOS reflects an allo-immunological injury, possibly triggered by cytomegalovirus and respiratory viral infections, or noninfectious injury. Immunological susceptibility may be reflected by more frequent acute rejection episodes. Preventative and therapeutic modifications in immunosuppression remain important. Identifying markers of immunological susceptibility and, hence, risk stratification requires further research. PMID- 15819591 TI - Optimising the use of beta-adrenoceptor antagonists in coronary artery disease. AB - beta-Adrenoceptor antagonists (beta-blockers) provide multiple benefits to patients with coronary artery disease. The 2001 American Heart Association and American College of Cardiology (AHA/ACC) guidelines for secondary prevention of myocardial infarction (MI) recommend initiating beta-adrenoceptor blockade in all post-MI patients and continuing therapy indefinitely. Atenolol and metoprolol have been shown to decrease vascular mortality in the acute-MI period. In the post-MI period timolol provided a 39% reduction in mortality in the Norwegian Multicenter Study group and propranolol was associated with a 26% reduction in mortality in BHAT (Beta-blocker Heart Attack Trial). beta-Adrenoceptor antagonist therapy results in reduction of myocardial oxygen demand and is therefore also effective for the treatment of angina pectoris. In CAST (Cardiac Arrhythmia Suppression Trial) beta-adrenoceptor antagonist therapy was associated with a significant reduction in arrhythmic death or cardiac arrest. In the post-MI amiodarone trials EMIAT (European Myocardial Infarct Amiodarone Trial) and CAMIAT (Canadian Amiodarone Myocardial Infarction Trial) there was a mortality benefit and decreased arrhythmic death in patients who received both amiodarone and beta adrenoceptor antagonist therapy, compared with patients receiving amiodarone therapy alone. In the post-MI defibrillator (implantable cardioverter defibrillator [ICD]) trials, AVID (Antiarrhythmic Versus Implantable Defibrillator) and MUSTT (Multicenter Unsustained Tachycardia Trial), beta adrenoceptor antagonist therapy was independently associated with improved overall survival. The exception was the ICD patients in MUSTT, and the benefit was attenuated in the amiodarone and ICD patients in AVID.AHA/ACC guidelines recommend the use of beta-adrenoceptor antagonists in all patients with symptomatic left ventricular dysfunction, based on several large, controlled heart failure trials. Extended-release metoprolol succinate reduced all-cause mortality by 34% in MERIT-HF (Metoprolol Controlled-Release/Extended-Release Randomized Intervention Trial in Heart Failure). Bisoprolol was associated with a 34% mortality benefit in CIBIS-II (Cardiac Insufficiency Bisoprolol Study II) and carvedilol was associated with a 35% mortality reduction in the COPERNICUS (Carvedilol Prospective Randomized Cumulative Survival) trial. beta-Adrenoceptor antagonists reduce perioperative mortality in patients undergoing cardiac as well as non-cardiac surgery; however, they remain underutilised. Contraindications to beta-adrenoceptor antagonist therapy include severe bradycardia, high-grade atrioventricular block, marked sinus node dysfunction and acute exacerbations of heart failure. Many of the perceived adverse effects of beta-adrenoceptor antagonists have not been substantiated by large clinical trials.beta Adrenoceptor antagonists differ with regard to receptor selectivity, receptor affinity, lipophilicity and intrinsic sympathomimetic activity. Beneficial properties of beta-adrenoceptor antagonists may not always be extrapolated as a class effect, and patient selection and drug preparations should follow trial guidelines. The beneficial effects of beta-adrenoceptor antagonists are clearly proven in cardiac patients and those at risk for cardiac disease. They are indicated for heart failure and proven beneficial in patients undergoing cardiac and non-cardiac surgery. These benefits appear to be consistent across most patient subgroups. beta-Adrenoceptor antagonists are generally well tolerated, yet significant morbidity and mortality result from their continued underutilisation. PMID- 15819590 TI - Sexual dysfunction in male patients with hypertension: influence of antihypertensive drugs. AB - Evidence suggests that arterial hypertension, in addition to being a cardiovascular and renal risk factor, may also be associated with an impairment of male sexual function. Since other cardiovascular risk factors, especially diabetes mellitus, have also been shown to correlate with impaired sexual function it has been proposed that sexual and especially erectile dysfunction may, at least in part, represent just another manifestation of atherosclerotic vascular disease. In addition to hypertension itself, sexual function in male hypertensive patients may also be affected by antihypertensive drug treatment. Available evidence suggests that centrally acting sympatholytic agents, beta adrenoceptor antagonists (beta-blockers) and diuretics may have the potential to further impair sexual function. Calcium channel antagonists and ACE inhibitors may be neutral with respect to this endpoint. Preliminary data from several randomised and open studies have suggested that angiotensin II (AT)(1)-receptor antagonists may even be associated with an improvement of sexual function. However, many aspects of the interaction between hypertension, antihypertensive drug treatment and male sexual function remain unclear. Among other factors, the relative contribution of disease labelling both to the higher incidence of sexual dysfunction in hypertensive versus normotensive males and to the negative impact of treatment remains an open question. Furthermore, dose dependence of the observed effects of antihypertensive agents on sexual function, the role of combination therapy and the anticipation of proposed adverse effects of treatment are unresolved issues. Thus, more data from studies of high quality using standardised definitions and procedures are urgently needed to at least partially resolve some of the many open questions. PMID- 15819592 TI - Mycophenolate sodium delayed release: prevention of renal transplant rejection. AB - Mycophenolate sodium delayed release is an enteric-coated formulation designed to release the active agent (mycophenolic acid) in the small intestine. Mycophenolate sodium delayed release was equivalent in efficacy to, and had a similar tolerability profile as, mycophenolate mofetil in de novo renal transplant patients treated with ciclosporin (cyclosporin) and corticosteroids in a well designed study of 12 month's duration. Maintenance renal transplant patients treated with ciclosporin and corticosteroids were converted from mycophenolate mofetil to mycophenolate sodium delayed release without any alteration in efficacy or tolerability in a well designed study of 12 month's duration. The incidence of gastrointestinal adverse events with mycophenolate sodium delayed release was similar to that with mycophenolate mofetil. PMID- 15819595 TI - Peginterferon-alpha-2a (40kD): a review of its use in the management of patients with chronic hepatitis B. AB - Subcutaneous peginterferon-alpha-2a (40kD) [Pegasys] is an effective and reasonably well tolerated treatment for the management of patients with hepatitis B e antigen (HBeAg)-negative or -positive chronic hepatitis B. It was significantly more effective than lamivudine monotherapy at inducing sustained virological response and ALT normalisation in both HBeAg-negative and -positive patients. Notably, the addition of lamivudine to peginterferon-alpha-2a (40kD) conferred no additional benefit versus peginterferon-alpha-2a (40kD) monotherapy. Moreover, in HBeAg-positive patients significantly more peginterferon-alpha-2a (40kD) recipients experienced HBeAg seroconversion than lamivudine recipients. The position of peginterferon-alpha-2a (40kD) relative to other treatment options remains to be fully determined. In the meantime, this agent appears to be a valuable new option for the management of patients with HBeAg-negative or positive chronic hepatitis B. PMID- 15819597 TI - Valganciclovir: a review of its use in the management of CMV infection and disease in immunocompromised patients. AB - Valganciclovir (Valcyte) is an orally administered prodrug of the standard anti cytomegalovirus (CMV) drug ganciclovir. Valganciclovir is as effective as intravenous ganciclovir for the treatment of AIDS-related CMV retinitis, and oral ganciclovir for the prophylaxis of CMV infection and disease in high-risk solid organ transplant recipients. The drug is generally well tolerated and has a similar tolerability profile to that of oral or intravenous ganciclovir, but is devoid of adverse events related to intravenous or indwelling catheter access associated with the use of intravenous ganciclovir, cidofovir and foscarnet. The simple and convenient once-daily valganciclovir regimen offers potential for improved patient compliance. It provides greater systemic ganciclovir exposure than oral ganciclovir, thus reducing the risk of viral resistance when used for prophylaxis in high-risk solid organ transplant recipients. Furthermore, the use of valganciclovir instead of intravenous ganciclovir may provide significant cost savings, based on data comparing oral versus intravenous regimens for the treatment of AIDS-related CMV retinitis. Overall, valganciclovir appears to have some advantages over ganciclovir. Therefore, when used as prophylaxis against CMV infection and disease in high-risk solid organ transplant recipients or as induction and maintenance therapy of CMV retinitis in patients with AIDS, oral valganciclovir is an attractive alternative to other available anti-CMV drugs. PMID- 15819598 TI - Literacy and health outcomes. PMID- 15819596 TI - Tacrolimus ointment: a review of its use in atopic dermatitis and its clinical potential in other inflammatory skin conditions. AB - Tacrolimus ointment (Protopic) is a topically applied macrolide lactone immunomodulator effective in the treatment of atopic dermatitis. Its mechanism of action primarily involves calcineurin inhibition, which interrupts cytokine gene expression and leads to the downregulation of T-cell activity. Tacrolimus ointment (0.03% and 0.1% for adults and 0.03% for children) is an effective treatment for atopic dermatitis of the trunk and limbs, as well as sensitive skin areas such as the face. Its efficacy is similar to or greater than that of hydrocortisone acetate 1%, hydrocortisone butyrate 0.1% and betamethsone valerate 0.12% ointments and pimecrolimus 1% cream. Systemic absorption of tacrolimus from the ointment is minimal, and adverse events, which are mostly associated with the application site and include skin burning and pruritus, tend to resolve early in treatment. Unlike topical corticosteroids, tacrolimus ointment is not associated with skin atrophy, and it is a well tolerated treatment for adults or children with atopic dermatitis, particularly when long-term treatment is indicated or the face or skin-fold regions are involved. PMID- 15819599 TI - UV doses worldwide. AB - UV radiation affects human health. Human exposure to UV radiation causes a few beneficial health effects like vitamin D3 formation but it causes many detrimental health effects: sunburn, ocular damage, photoaging, immune suppression, DNA damage and skin cancer. In countries with fair-skinned populations, skin cancer is the most diagnosed of all cancers. In the United States in 2002, there were over one million new skin cancer cases. That means one out of every 285 people got skin cancer. Skin cancer of fair-skinned individuals is increasing at an alarming rate (4-6% per year) around the world and has now reached so-called "pandemic" proportions. Thus, it is important to know what UV doses people around the world get throughout their lives. This review covers how the outdoor UV doses are weighted for different biological effects, the most commonly used measuring devices for terrestrial and personal UV doses, the natural and other effects on terrestrial and personal UV doses, the time people spend outside, their ambient exposures and the terrestrial and personal UV doses of adult outdoor and indoor workers as well as children and adolescents around the world. Overall, outdoor-working adults get about 10%, while indoor-working adults and children get about 3% (2-4%) of the total available annual UV (on a horizontal plane). People's UV doses increase with increasing altitude and decreasing latitude; most indoor-working adult Europeans get 10,000-20,000 J/m2 per year, Americans get 20,000-30,000 J/m2 per year and Australians are estimated to get 20,000-50,000 J/m2 per year (excluding vacation, which can increase the dose by 30% or more). PMID- 15819600 TI - The use of wavelength-selective plastic cladding materials in horticulture: understanding of crop and fungal responses through the assessment of biological spectral weighting functions. AB - Plant responses to light spectral quality can be exploited to deliver a range of agronomically desirable end points in protected crops. This can be achieved using plastics with specific spectral properties as crop covers. We have studied the responses of a range of crops to plastics that have either (a) increased transmission of UV compared with standard horticultural covers, (b) decreased transmission of UV or (c) increased the ratio of red (R) : far-red (FR) radiation. Both the UV-transparent and R : FR increasing films reduced leaf area and biomass, offering potential alternatives to chemical growth regulators. The UV-opaque film increased growth, but while this may be useful in some crops, there were trade-offs with elements of quality, such as pigmentation and taste. UV manipulation may also influence disease control. Increasing UV inhibited not only the pathogenic fungus Botrytis cinerea but also the disease biocontrol agent Trichoderma harzianum. Unlike B. cinerea, T. harzianum was highly sensitive to UV A radiation. These fungal responses and those for plant growth in the growth room and the field under different plastics are analyzed in terms of alternative biological spectral weighting functions (BSWF). The role of BSWF in assessing general patterns of response to UV modification in horticulture is also discussed. PMID- 15819601 TI - Virus particles and receptor interaction monitored by fluorescence spectroscopy. AB - Native fluorescence spectroscopy (NFS), primarily from tryptophan (trp), was used for in situ investigation of the virus-receptor attachment process in phi6, a lipid-containing bacteriophage from the Cystoviridae family. NFS allowed us to monitor the viral attachment directly to its receptor, which was isolated from the pseudomonad host. Immediately upon mixing, an increase in tryptophan emission intensity was observed followed by a subsequent decrease in emission intensity. The initial increase in emission intensity reflects changes in trp quantum efficiency as the phi6 surface proteins change their conformation as a result of virus attachment to the pilus. The cystovirus spike protein P3 is responsible for receptor recognition and the fluorescence changes observed are likely to be the consequence of its conformational transition at this initial infection stage, providing a kinetic view of this process. The subsequent decrease in trp emission intensity could be due to changes in viral proteins as a result of disassembly of the pili. The technique may have important applications for the dynamic monitoring of additional stages of the virus replication cycle such as assembly, interaction with nucleic acids and maturation. This work expands on a previous demonstration that fluorescence offered a novel tool to detect virus particle interaction with its host cell. PMID- 15819602 TI - Protection by food-derived antioxidants from UV-A1-induced photodamage, measured using living skin equivalents. AB - In a study of biomarkers of ultraviolet-A1 radiation (UV-A1)-induced skin damage, living skin equivalent cultures (LSE) were treated with the antioxidants hesperetin and quercetin-3-glucoside and irradiated with 25 or 50 J/cm2 UV-A1. Changes in the following biomarkers were measured; Interleukin 1-alpha (IL 1alpha), Heme Oxygenase-1 (HO-1), TdT-mediated dUTP nick end labeling (TUNEL) and 8-hydroxy-2'-deoxyguanosine (8-OHdG). IL-1alpha and HO-1 were analyzed by real time PCR, Western blot, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. TUNEL and 8-OHdG were determined by (immuno)histochemical techniques. Sections were stained with hematoxylin and eosin (H&E). UV-A1 induced keratinocyte and fibroblast vacuolation and nuclear pyknosis, intense TUNEL staining of fibroblasts and increased staining of cells and nuclei for 8-OHdG. Lesser or marginal increases in intensity followed staining for HO-1 and IL 1alpha. The IL-1alpha increase was confirmed by ELISA assays of the medium supernatants. Hesperetin and quercetin-3-glucoside reduced changes in H&E, 8 OHdG, TUNEL and IL-1alpha. Quercetin-3-glucoside reduced the amount of IL-1alpha in LSE media. These observations support the use of the selected biomarkers to monitor UV-A1 damage and provide evidence that dietary ingredients could reduce ultraviolet-A radiation-induced damage. PMID- 15819603 TI - Effect of digitonin on the rhodopsin meta I-meta II equilibrium. AB - Absorbance difference spectra were recorded from 10 micros to 540 ms after photoexcitation of sonicated suspensions of hypotonically washed bovine rod outer segments with varying amounts of the detergent digitonin added (0 to 2%) at 20 degrees C. Metarhodopsin I480 and metarhodopsin II displayed the expected anomalous pH dependence at pH 6 and 8 (i.e. opposite to that expected from direct protonation of the chromophore Schiff base). However, increasing levels of digitonin eliminated the pH dependence of the equilibrium, and at 2% digitonin the pH 6 and pH 8 data were both similar to the data collected at pH 8 without digitonin. Addition of 0.5% azolectin restored approximately 50% of the anomalous pH dependence at pH 6 in the 2% digitonin sample. The possibility that digitonin induced large-scale aggregation of rhodopsin in the disk membrane that could be reversed by azolectin was tested using time-resolved linear dichroism. Those results showed that even 0.3% digitonin disrupted the membrane, and no large aggregates were detectable under any conditions. Thus, digitonin reduces the activity of a component of the disk membrane required for metarhodopsin II formation, and that deficiency can be compensated for by azolectin. PMID- 15819605 TI - As the antarctic ice pack recedes, a fragile ecosystem hangs in the balance. PMID- 15819604 TI - The stability of a stochastic CaMKII switch: dependence on the number of enzyme molecules and protein turnover. AB - Molecular switches have been implicated in the storage of information in biological systems. For small structures such as synapses, these switches are composed of only a few molecules and stochastic fluctuations are therefore of importance. Such fluctuations could potentially lead to spontaneous switch reset that would limit the lifetime of information storage. We have analyzed a model of the calcium/calmodulin-dependent protein kinase II (CaMKII) switch implicated in long-term memory in the nervous system. The bistability of this switch arises from autocatalytic autophosphorylation of CaMKII, a reaction that is countered by a saturable phosphatase-1-mediated dephosphorylation. We sought to understand the factors that control switch stability and to determine the functional relationship between stability and the number of molecules involved. Using Monte Carlo simulations, we found that the lifetime of states of the switch increase exponentially with the number of CaMKII holoenzymes. Switch stability requires a balance between the kinase and phosphatase rates, and the kinase rate must remain high relative to the rate of protein turnover. Thus, a critical limit on switch stability is set by the observed turnover rate (one per 30 h on average). Our computational results show that, depending on the timescale of fluctuations in enzyme numbers, for a switch composed of about 15 CaMKII holoenzymes, the stable persistent activation can span from a few years to a human lifetime. PMID- 15819606 TI - The PLoS community journals. AB - 2005 will see the launch of three new open access journals from the Public Library of Science - PLoS Computational Biology, PLoS Genetics, and PLoS Pathogens. PMID- 15819607 TI - Alchemy and the new age of cardiac muscle cell biology. PMID- 15819608 TI - Perception space--the final frontier. PMID- 15819612 TI - Overexpression of truncated gamma-tubulins disrupts mitotic aster formation in Xenopus oocyte extracts. AB - Mechanisms of spindle pole formation rely on minus-end-directed motor proteins. gamma-Tubulin is present at the centre of poles, but its function during pole formation is completely unknown. To address the role of gamma-tubulin in spindle pole formation, we overexpressed GFP (green fluorescent protein)-fused gamma tubulin (gamma-Tu-GFP) in Xenopus oocytes and produced self-assembled mitotic asters in the oocyte extracts. gamma-Tu-GFP associated with endogenous alpha-, beta- and gamma-tubulin, suggesting that it acts in the same manner as that of endogenous gamma-tubulin. During the process of aster formation, gamma-Tu-GFP aggregated as dots on microtubules, and then the dots were translocated to the centre of the aster along microtubules in a manner dependent on cytoplasmic dynein activity. Inhibition of the function of gamma-tubulin by an anti-gamma tubulin antibody resulted in failure of microtubule organization into asters. This defect was restored by overexpression of gamma-Tu-GFP, confirming the necessity of gamma-tubulin in microtubule recruitment for aster formation. We also examined the effects of truncated gamma-tubulin mutants, which are difficult to solubly express in other systems, on aster formation. The middle part of gamma tubulin caused abnormal organization of microtubules in which minus ends of microtubules were not tethered, but dispersed. An N-terminus-deleted mutant prevented recruitment of microtubules into asters, similar to the effect of the anti-gamma-tubulin antibody. The results indicate possible roles of gamma-tubulin in spindle pole formation and show that the system developed in the present study could be useful for analysing roles of many proteins that are difficult to solubly express. PMID- 15819609 TI - Ongoing spontaneous activity controls access to consciousness: a neuronal model for inattentional blindness. AB - Even in the absence of sensory inputs, cortical and thalamic neurons can show structured patterns of ongoing spontaneous activity, whose origins and functional significance are not well understood. We use computer simulations to explore the conditions under which spontaneous activity emerges from a simplified model of multiple interconnected thalamocortical columns linked by long-range, top-down excitatory axons, and to examine its interactions with stimulus-induced activation. Simulations help characterize two main states of activity. First, spontaneous gamma-band oscillations emerge at a precise threshold controlled by ascending neuromodulator systems. Second, within a spontaneously active network, we observe the sudden "ignition" of one out of many possible coherent states of high-level activity amidst cortical neurons with long-distance projections. During such an ignited state, spontaneous activity can block external sensory processing. We relate those properties to experimental observations on the neural bases of endogenous states of consciousness, and particularly the blocking of access to consciousness that occurs in the psychophysical phenomenon of "inattentional blindness," in which normal subjects intensely engaged in mental activity fail to notice salient but irrelevant sensory stimuli. Although highly simplified, the generic properties of a minimal network may help clarify some of the basic cerebral phenomena underlying the autonomy of consciousness. PMID- 15819613 TI - The spot 14 protein inhibits growth and induces differentiation and cell death of human MCF-7 breast cancer cells. AB - The S14 (spot 14) gene encodes a protein that is predominantly expressed in lipogenic tissues, such as the liver, white and brown adipose tissues and the lactating mammary glands. Accumulated evidence suggests that S14 could play an important role in the induction of lipogenic enzymes. In humans, the S14 locus resides in the chromosome region 11q13, which is frequently amplified in breast tumours, and as a result, it has been suggested that this protein could play a role in the metabolism and growth of these kinds of tumours. In the present study, we have examined the effects of S14 overexpression in MCF-7 human breast cancer cells. We found that S14 causes (i) an inhibition of cell proliferation and of anchorage-independent growth, (ii) a marked reduction in the number of viable cells and (iii) the induction of differentiation and cell death of these cells. The inhibition of cell growth was associated with a decrease in the expression of cyclin D1 and a reduction of cyclin D1 promoter activity. Increased expression of S14 also caused the accumulation of cytochrome c in the cytosol and loss of mitochondrial membrane potential. These findings suggest that S14 may function as an important modulator of tumorigenesis in human breast by decreasing cell growth and inducing cell death and differentiation. PMID- 15819614 TI - Transforming growth factor-beta receptors localize to caveolae and regulate endothelial nitric oxide synthase in normal human endothelial cells. AB - Caveolae (sphingolipid- and cholesterol-rich, 100 nm flask-shaped invaginations of the cell membrane) serve as a nexus of cell signalling. In the present study caveolin-rich lipid raft domains were extracted from HUVEC (human umbilical-vein endothelial cells) using both density gradient and immunoprecipitation techniques, and demonstrated localization of the TGF-beta (transforming growth factor-beta) receptors TbetaRI and TbetaRII to the Cav-1 (caveolin-1)-enriched raft fractions of these normal, human endothelial cells. Immunoprecipitation demonstrated an association between TbetaRI and TbetaRII, as well as an association of the TbetaRs receptors with Cav-1 and eNOS (endothelial nitric oxide synthase), suggesting a mutual co-localization to caveolae; after treatment of HUVEC with 5 ng/ml TGF-beta1 for 15 min, however, co-precipitation of eNOS with TbetaRI, TbetaRII and Cav-1 was diminished. The loss of immunoprecipitable eNOS from Cav-1-enriched fractions was accompanied by a decrease both in phosphorylation of eNOS and in enzymatic activity (conversion of arginine into citrulline). No change in the localization of eNOS to morphologically distinct caveolae could be detected by electron microscopy after treatment of HUVEC with TGF-beta1 for 20 min. The results of these investigations provide evidence that TbetaRI interacts with eNOS in the caveolae of normal, human endothelial cells and has a regulatory function on basal eNOS enzymatic activity. PMID- 15819615 TI - Oiling the key hole. AB - Many bacteria have been found to interact with specialized domains, rich in cholesterol and sphingolipids, of the host plasma membrane, termed lipid rafts. The mechanisms that underlie this interaction are starting to be unravelled. In this issue, Hayward et al. show that early effector proteins secreted by type III secretion harbouring Gram-negative bacteria are in fact cholesterol-binding proteins. Combined with other recent findings, this work shows that multiple steps leading to infection by these bacteria depend on raft components: activation of secretion, binding, perforation of the host cell membrane and signalling to trigger bacterial engulfment. PMID- 15819616 TI - Differential gene expression in genetically identical sister cells: the initiation of sporulation in Bacillus subtilis. AB - Early in sporulation, the cell divides asymmetrically to give two sister compartments, a smaller prespore and a larger mother cell. Differential gene expression in these compartments depends on the regulation of the first sporulation-specific sigma factor, sigma(F), which is activated only in the prespore. Regulation relies on the interactions of four proteins -sigma(F), its antisigma SpoIIAB (which also has protein kinase activity), the anti-antisigma SpoIIAA and the protein phosphatase SpoIIE. Before asymmetric division, and in the mother cell after division, sigma(F) is held in an inactive complex with SpoIIAB and ATP; SpoIIAA is in its phosphorylated form. To disrupt the complex so as to liberate sigma(F) in the prespore, dephosphorylated SpoIIAA is needed, and this is made available by SpoIIE. Thereafter, SpoIIAB and SpoIIE are active simultaneously in the prespore, cycling SpoIIAA through phosphorylated and non phosphorylated forms. This cycle detains SpoIIAB in a state in which it cannot inhibit sigma(F). Results from biophysical techniques, mathematical simulations and enzyme kinetics have now helped to elucidate the dynamics of the protein protein interactions involved. An understanding of these dynamics largely accounts for the regulation of sigma(F). We show that the system is tuned to be highly efficient in its use of components and extremely economical in conserving ATP. PMID- 15819617 TI - Cholesterol binding by the bacterial type III translocon is essential for virulence effector delivery into mammalian cells. AB - A ubiquitous early step in infection of man and animals by enteric bacterial pathogens like Salmonella, Shigella and enteropathogenic Escherichia coli (EPEC) is the translocation of virulence effector proteins into mammalian cells via specialized type III secretion systems (TTSSs). Translocated effectors subvert the host cytoskeleton and stimulate signalling to promote bacterial internalization or survival. Target cell plasma membrane cholesterol is central to pathogen-host cross-talk, but the precise nature of its critical contribution remains unknown. Using in vitro cholesterol-binding assays, we demonstrate that Salmonella (SipB) and Shigella (IpaB) TTSS translocon components bind cholesterol with high affinity. Direct visualization of cell-associated fluorescently labelled SipB and parallel immunogold transmission electron microscopy revealed that cholesterol levels limit both the amount and distribution of plasma membrane integrated translocon. Correspondingly, cholesterol depletion blocked effector translocation into cultured mammalian cells by not only the related Salmonella and Shigella TTSSs, but also the more divergent EPEC system. The data reveal that cholesterol-dependent association of the bacterial TTSS translocon with the target cell plasma membrane is essential for translocon activation and effector delivery into mammalian cells. PMID- 15819618 TI - Stripping Bacillus: ComK auto-stimulation is responsible for the bistable response in competence development. AB - In Bacillus subtilis competence for genetic transformation develops only in a subpopulation of cells in an isogenic culture. The molecular mechanisms underlying this phenotypic heterogeneity are unknown. In this study, we stepwise simplify the signal transduction cascade leading to competence, yielding a strain devoid of all regulatory inputs for this process that have been identified so far. We demonstrate that auto-stimulation of ComK, the master regulator for competence development, is essential and in itself can be sufficient to generate a bistable expression pattern. We argue that transcriptional regulation determines the threshold of ComK to initiate the auto-stimulatory response, and that the basal level of ComK (in a wild-type strain governed by MecA-mediated proteolytic control) determines the fraction of cells that reach this threshold, and thus develop competence. PMID- 15819619 TI - Bistability in the Bacillus subtilis K-state (competence) system requires a positive feedback loop. AB - High expression of the transcriptional activator ComK occurs in 10-20% of the cells in stationary phase cultures of Bacillus subtilis strain 168. ComK drives the expression of more than 100 genes constituting the semidormant K-state, distinct from sporulation and vegetative growth. Among the genes so activated are those that permit competence for genetic transformation. We have addressed the origin of bistability in expression of ComK. We show that bistability requires positive autoregulation at the promoter of comK, but not a potential toggle switch, in which ComK represses the promoter of rok and Rok represses the promoter of comK. We further address the source of the noise that results in the stochastic selection of cells that will express comK. A revised model for the regulation of comK expression is proposed that partially explains bistability. PMID- 15819620 TI - Promoter architecture and response to a positive regulator of archaeal transcription. AB - The archaeal transcription apparatus is chimeric: its core components (RNA polymerase and basal factors) closely resemble those of eukaryotic RNA polymerase II, but the putative archaeal transcriptional regulators are overwhelmingly of bacterial type. Particular interest attaches to how these bacterial-type effectors, especially activators, regulate a eukaryote-like transcription system. The hyperthermophilic archaeon Methanocaldococcus jannaschii encodes a potent transcriptional activator, Ptr2, related to the Lrp/AsnC family of bacterial regulators. Ptr2 activates rubredoxin 2 (rb2) transcription through a bipartite upstream activating site (UAS), and conveys its stimulatory effects on its cognate transcription machinery through direct recruitment of the TATA binding protein (TBP). A functional dissection of the highly constrained architecture of the rb2 promoter shows that a 'one-site' minimal UAS suffices for activation by Ptr2, and specifies the required placement of this site. The presence of such a simplified UAS upstream of the natural rubrerythrin (rbr) promoter also suffices for positive regulation by Ptr2 in vitro, and TBP recruitment remains the primary means of transcriptional activation at this promoter. PMID- 15819621 TI - A bZIP transcription factor from Phytophthora interacts with a protein kinase and is required for zoospore motility and plant infection. AB - Zoospores are critical in the disease cycle of Phytophthora infestans, a member of the oomycete group of fungus-like microbes and the cause of potato late blight. A protein kinase induced during zoosporogenesis, Pipkz1, was shown to interact in the yeast two-hybrid system with a putative bZIP transcription factor. This interaction was confirmed in vitro using a pull-down assay. The transcription factor gene, Pibzp1, was single copy and expressed in all tissues. Transformants of P. infestans stably silenced for Pibzp1 were generated using plasmids expressing its coding region in sense or antisense orientations. A protoplast transformation method induced silencing more efficiently than transformation by an electroporation scheme. Wild-type and silenced strains exhibited no differences in hyphal growth or morphology, mating, sporangia production or zoospore release. However, zoospores from the mutants spun in tight circles, instead of exhibiting the normal pattern of straight swimming punctuated by turns. Zoospore encystment was unaffected by silencing, but cysts germinated more efficiently than controls. Germinated cysts from the mutants failed to develop appressoria and were unable to infect plants; however, they could colonize wounded tissue. These phenotypes indicate that Pibzp1 is a key regulator of several stages of the zoospore-mediated infection pathway. PMID- 15819622 TI - The Mep2p ammonium permease controls nitrogen starvation-induced filamentous growth in Candida albicans. AB - Nitrogen starvation is one of the signals that induce Candida albicans, the major fungal pathogen of humans, to switch from yeast to filamentous growth. In response to nitrogen starvation, C. albicans expresses the MEP1 and MEP2 genes, which encode two ammonium permeases that enable growth when limiting concentrations of ammonium are the only available nitrogen source. In addition to its role as an ammonium transporter, Mep2p, but not Mep1p, also has a central function in the induction of filamentous growth on a solid surface under limiting nitrogen conditions. When ammonium is absent or present at low concentrations, Mep2p activates both the Cph1p-dependent mitogen-activated protein (MAP) kinase pathway and the cAMP-dependent signalling pathway in a Ras1p-dependent fashion via its C-terminal cytoplasmic tail, which is essential for signalling but dispensable for ammonium transport. In contrast, under ammonium-replete conditions that require transporter-mediated uptake Mep2p is engaged in ammonium transport and signalling is blocked such that C. albicans continues to grow in the budding yeast form. Mep2p is a less efficient ammonium transporter than Mep1p and is expressed at much higher levels, a distinguishing feature that is important for its signalling function. At sufficiently high concentrations, ammonium represses filamentous growth even when the signalling pathways are artificially activated. Therefore, C. albicans has established a regulatory circuit in which a preferred nitrogen source, ammonium, also serves as an inhibitor of morphogenesis that is taken up into the cell by the same transporter that mediates the induction of filamentous growth in response to nitrogen starvation. PMID- 15819623 TI - Budding yeast PDS5 plays an important role in meiosis and is required for sister chromatid cohesion. AB - Budding yeast PDS5 is an essential gene in mitosis and is required for chromosome condensation and sister chromatid cohesion. Here we report that PDS also is required in meiosis. Pds5p localizes on chromosomes at all stages during meiotic cycle, except anaphase I. PDS5 plays an important role at first meiotic prophase. Failure in function of PDS5 causes premature separation of chromosomes. The loading of Pds5p onto chromosome requires the function of REC8, but the association of Rec8p with chromosome is independent of PDS5. Mutant analysis and live cell imaging indicate that PDS5 play a role in meiosis II as well. PMID- 15819624 TI - Mutational analysis of the group A streptococcal operon encoding streptolysin S and its virulence role in invasive infection. AB - The pathogen group A Streptococcus (GAS) produces a wide spectrum of infections including necrotizing fasciitis (NF). Streptolysin S (SLS) produces the hallmark beta-haemolytic phenotype produced by GAS. The nine-gene GAS locus (sagA-sagI) resembling a bacteriocin biosynthetic operon is necessary and sufficient for SLS production. Using precise, in-frame allelic exchange mutagenesis and single-gene complementation, we show sagA, sagB, sagC, sagD, sagE, sagF and sagG are each individually required for SLS production, and that sagE may further serve an immunity function. Limited site-directed mutagenesis of specific amino acids in the SagA prepropeptide supports the designation of SLS as a bacteriocin-like toxin. No significant pleotrophic effects of sagA deletion were observed on M protein, capsule or cysteine protease production. In a murine model of NF, the SLS-negative M1T1 GAS mutant was markedly diminished in its ability to produce necrotic skin ulcers and spread to the systemic circulation. The SLS toxin impaired phagocytic clearance and promoted epithelial cell cytotoxicity, the latter phenotype being enhanced by the effects of M protein and streptolysin O. We conclude that all genetic components of the sag operon are required for expression of functional SLS, an important virulence factor in the pathogenesis of invasive M1T1 GAS infection. PMID- 15819625 TI - Constitutive expression of yeast phospholipid biosynthetic genes by variants of Ino2 activator defective for interaction with Opi1 repressor. AB - Regulated expression of structural genes involved in yeast phospholipid biosynthesis is mediated by inositol/choline-responsive element (ICRE) upstream motifs, bound by the heterodimeric activator complex Ino2 + Ino4. Gene repression occurs in the presence of sufficient inositol and choline, requiring an intact Opi1 repressor which binds to Ino2. For a better understanding of interactions among regulators, we mapped an 18 aa repressor interaction domain (RID, aa 118 135) within Ino2 necessary and sufficient for binding by Opi1. By alanine scanning mutagenesis of the entire RID we were able to identify nine residues critical for Opi1-dependent repression of Ino2 function. Consequently, the corresponding dominant Ino2 variants conferred constitutive expression of an ICRE dependent reporter gene and were no longer inhibited even by overproduction of Opi1. Interestingly, Ino2 RID partially overlaps with transcriptional activation domain TAD2. As certain mutations exclusively affect repression while others affect both repression and activation, both functions of Ino2 can be functionally uncoupled. Correspondingly, we mapped the RID-binding activator interaction domain (AID, aa 321-380) at the C-terminus of Opi1 and introduced missense mutations at selected positions. An Opi1 variant simultaneously mutated at three highly conserved positions showed complete loss of repressor function, confirming RID-AID interaction as the crucial step of regulated expression of ICRE-dependent genes. PMID- 15819626 TI - Control of speed modulation (chemokinesis) in the unidirectional rotary motor of Sinorhizobium meliloti. AB - Swimming cells of Sinorhizobium meliloti are driven by flagella that rotate only clockwise. They can modulate rotary speed (achieve chemokinesis) and reorient the swimming path by slowing flagellar rotation. The flagellar motor is energized by proton motive force, and torque is generated by electrostatic interactions at the rotor/stator (FliG/MotA-MotB) interface. Like the Escherichia coli flagellar motor that switches between counterclockwise and clockwise rotation, the S. meliloti rotary motor depends on electrostatic interactions between conserved charged residues, namely, Arg294 and Glu302 (FliG) and Arg90, Glu98 and Glu150 (MotA). Unlike in E. coli, however, Glu150 is essential for torque generation, whereas residues Arg90 and Glu98 are crucial for the chemotaxis-controlled variation of rotary speed. Substitutions of either Arg90 or Glu98 by charge neutralizing residues or even by their smaller, charge-maintaining isologues, lysine and aspartate, resulted in top-speed flagellar rotation and decreased potential to slow down in response to tactic signalling (chemokinesis-defective mutants). The data infer a novel mechanism of flagellar speed control by electrostatic forces acting at the rotor/stator interface. These features have been integrated into a working model of the speed-modulating rotary motor. PMID- 15819627 TI - SspA is required for acid resistance in stationary phase by downregulation of H NS in Escherichia coli. AB - The stringent starvation protein A (SspA) is a RNA polymerase-associated protein and is required for transcriptional activation of bacteriophage P1 late promoters. However, the role of SspA in gene expression in Escherichia coli is essentially unknown. In this work, we show that SspA is essential for cell survival during acid-induced stress. Apparently, SspA inhibits stationary-phase accumulation of H-NS, a global regulator which functions mostly as a repressor, thereby derepressing multiple stress defence systems including those for acid stress and nutrient starvation. Consequently, the gene expression pattern of the H-NS regulon is altered in the sspA mutant, leading to acid-sensitive and hypermotile phenotypes. Thus, our study indicates that SspA is a global regulator, which acts upstream of H-NS, and thereby plays an important role in the stress response of E. coli during stationary phase. In addition, our results indicate that the expression of the H-NS regulon is sensitive to small changes in the cellular level of H-NS, enabling the cell to response rapidly to environment cues. As SspA and H-NS are highly conserved among Gram-negative bacteria, of which many are pathogenic, the global role of SspA in the stress response and pathogenesis is discussed. PMID- 15819628 TI - Two acid-inducible promoters from Lactococcus lactis require the cis-acting ACiD box and the transcription regulator RcfB. AB - We previously characterized three Lactococcus lactis promoters, P170, P1 and P3, which are induced by low pH. Here, we identified a novel 14 bp regulatory DNA region centred at around -41.5 and composed of three tetranucleotide sequences, boxes A, C and D. Boxes A and C contribute to P1 activity, whereas box D and the position of boxes ACD (renamed ACiD-box) are essential to P1 activity and acid response. We also identified a trans -acting protein, RcfB, which is involved in P170 and P1 basal activity and is essential for their pH induction. The regulator belongs to the Crp-Fnr family of transcription regulators. Overexpression of rcfB resulted in increased beta-galactosidase activities and lantibiotic lacticin 481 production from P170- and P1-controlled genes, respectively, in acid condition. RcfB is thus probably activated when cells encounter an acid environment. rcfB is co-transcribed with genes encoding an universal stress-like protein and a multidrug transporter. RcfB plays a role in acid adaptation, as the survival rate of an rcfB mutant after a lethal acid challenge was 130-fold lower than that of the wild-type strain, when the bacteria were first grown in acidic medium. The groESL promoter includes a sequence resembling an ACiD-box and the chaperone GroEL production is partly RcfB dependent in acid condition. Our results suggest that the ACiD-box could be the DNA target site of RcfB. PMID- 15819629 TI - Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene. AB - Clostridium perfringens type A isolates carrying an enterotoxin (cpe) gene are an important cause of human gastrointestinal diseases, including food poisoning, antibiotic-associated diarrhoea (AAD) and sporadic diarrhoea (SD). Using polymerase chain reaction (PCR), the current study determined that the cpb2 gene encoding the recently discovered beta2 toxin is present in <15% of food poisoning isolates, which typically carry a chromosomal cpe gene. However, >75% of AAD/SD isolates, which usually carry a plasmid cpe gene, tested cpb2(+) by PCR. Western blot analysis demonstrated that >97% of those cpb2(+)/cpe(+) AAD/SD isolates can produce CPB2. Additional PCR analyses, sequencing studies and pulsed field gel electrophoresis experiments determined that AAD/SD isolates carry cpb2 and cpe on the same plasmid when IS1151 sequences are present downstream of cpe, but cpb2 and cpe are located on different plasmids in AAD/SD isolates where IS1470-like sequences are present downstream of cpe. Those analyses also demonstrated that two different CPB2 variants (named CPB2h1 or CPB2h2) can be produced by AAD/SD isolates, dependent on whether IS1470-like or IS1151 sequences are present downstream of their cpe gene. CPB2h1 is approximately 10-fold more cytotoxic for CaCo-2 cells than is CPB2h2. Collectively, these results suggest that CPB2 could be an accessory toxin in C. perfringens enterotoxin (CPE)-associated AAD/SD. PMID- 15819630 TI - The exbD gene of Photorhabdus temperata is required for full virulence in insects and symbiosis with the nematode Heterorhabditis. AB - Photorhabdus are bacteria found colonizing the gut of a specialized stage of the nematode Heterorhabditis, called the infective juvenile (IJ). The IJ is a free living stage of the nematode that seeks out and infects insect larvae. Once inside the insect the IJ release Photorhabdus into the haemolymph where the bacteria rapidly proliferate, killing the insect within 48-72 h. The nematodes grow and reproduce in the insect cadaver by feeding on the Photorhabdus biomass. In this study we use Photorhabdus temperata K122 to show that genes involved in iron acquisition play a key role during the course of the tripartite bacteria nematode-insect interaction. We show that a strain carrying a mutation in a gene with homology to exbD, encoding a component of the TonB complex, is unable to grow well in conditions where iron is not freely available. In addition, this mutant, BMM417, requires a longer time to kill the insect larvae than the wild type bacteria and this defect in pathogenicity is complemented by the co injection of iron. Moreover, the increase in LT(50) observed with BMM417 is correlated with a significantly slower in vivo growth rate suggesting that iron is limiting in the insect. We also show that BMM417 is unable to support the growth and development of the Heterorhabditis nematode. Addition of exogenous iron to the growth media restores nematode growth and development on BMM417, suggesting that aspects of iron metaboism in Photorhabdus are important during the symbiosis with the nematode. PMID- 15819631 TI - Broad analysis reveals a consistent pattern of var gene transcription in Plasmodium falciparum repeatedly selected for a defined adhesion phenotype. AB - Transcription of the majority of the members of the Plasmodium falciparum var multigene family were analysed in two isolates by a quantitative approach. Both of these isolates had been repeatedly selected for adhesion to chondroitin sulphate A (CSA) and one had also been selected for adhesion to hyaluronic acid (HA). These adhesion phenotypes are expressed by many parasites isolated from placentae and are associated with malaria disease in pregnancy. Increased transcription of the var gene var2csa, or its homologue IT4 var4, was associated with the CSA and HA adhesion phenotypes in all parasites suggesting that it was the dominant, if not the only, var gene that encoded adhesion to CSA in these allogeneic isolates. Some var genes were consistently transcribed at higher levels than others regardless of expressed adhesion phenotypes suggesting a transcriptional hierarchy. Unspliced or partial transcripts were detected for most var genes tested. These atypical var gene transcripts may have implications for the regulation of var gene transcription. PMID- 15819632 TI - The alternative sigma factor sigma is required for resistance of Salmonella enterica serovar Typhimurium to anti-microbial peptides. AB - The enteric pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) encounters a variety of anti-microbial peptides during the course of infection. We report here that the extracytoplasmic sigma factor sigma(E) (RpoE) is required for Salmonella resistance to killing by the bactericidal/permeability-increasing protein (BPI)-derived peptide P2 and the murine alpha-defensin cryptdin-4 (Crp4). Moreover, sigma(E)-deficient S. Typhimurium is attenuated for virulence after oral infection of immunocompromised gp91phox(-/-) mice that lack a functional NADPH phagocyte oxidase, suggesting that sigma(E) plays an important role in resistance to non-oxidative mucosal host defences such as anti-microbial peptides. Although both P2 and Crp4 target the cell envelope, bacterial killing by these peptides appears to occur by distinct mechanisms. Formate enhances bacterial resistance to P2, as previously demonstrated, but not to Crp4. Both sigma(E) and cytoplasmic membrane-associated formate dehydrogenase are required for the protective effect of formate against P2. In contrast to P2, Crp4 does not inhibit bacterial respiration at lethal concentrations. However, both peptides induce expression of rpoE, suggesting that they trigger a common mechanism for sensing extracytoplasmic stress. PMID- 15819633 TI - The initial step of glycerolipid metabolism in Leishmania major promastigotes involves a single glycerol-3-phosphate acyltransferase enzyme important for the synthesis of triacylglycerol but not essential for virulence. AB - The synthesis of the major phospholipids, including those that play an essential role in Leishmania virulence, initiates with the acylation of glycerol-3 phosphate and dihydroxyacetonephosphate at the sn-1 position by glycerol-3 phosphate and dihydroxyacetonephosphate acyltransferases respectively. In this study, we show that Leishmania major promastigotes express a single glycerol-3 phosphate acyltransferase activity important for triacylglycerol synthesis but not essential for virulence. The encoding gene, LmGAT, expressed in yeast results in full complementation of the lethality of a mutant, gat1Deltagat2Delta, lacking glycerol-3-phosphate activity. Biochemical analyses revealed that LmGAT is a low affinity glycerol-3-phosphate acyltransferase and exhibits higher specific activity with unsaturated long fatty acyl-CoA donors. A L. major null mutant, Deltalmgat/Deltalmgat, was created and a thorough analysis of its lipid composition was performed. Deletion of LmGAT resulted in a complete loss of Leishmania glycerol-3-phosphate acyltransferase activity and a major reduction in triacylglycerol synthesis. Consistent with the specificity of LmGAT for glycerol 3-phosphate but not dihydroxyacetonephosphate, Deltalmgat/Deltalmgat mutant expressed normal levels of the ether-lipid derivatives and virulence factors, lipophosphoglycan and GPI-anchored proteins, gp63, and its virulence was not affected in mice. PMID- 15819634 TI - Alternative sigma factor interactions in Salmonella: sigma and sigma promote antioxidant defences by enhancing sigma levels. AB - Hierarchical interactions between alternative sigma factors control sequential gene expression in Gram-positive bacteria, whereas alternative sigma factors in Gram-negative bacteria are generally regarded to direct expression of discrete gene subsets. In Salmonella enterica serovar Typhimurium (S. Typhimurium), sigma(E) responds to extracytoplasmic stress, whereas sigma(H) responds to heat shock and sigma(S) is induced during nutrient limitation. Deficiency of sigma(E), sigma(H) or sigma(S) increases S. Typhimurium susceptibility to oxidative stress, but an analysis of double and triple mutants suggested that antioxidant actions of sigma(E) and sigma(H) might be dependent on sigma(S). Transcriptional profiling of mutant Salmonella lacking sigma(E) revealed reduced expression of genes dependent on sigma(H) and sigma(S) in addition to sigma(E). Further investigation demonstrated that sigma(E) augments sigma(S) levels during stationary phase via enhanced expression of sigma(H) and the RNA-binding protein Hfq, leading to increased expression of sigma(S)-dependent genes and enhanced resistance to oxidative stress. Maximal expression of the sigma(S)-regulated gene katE required sigma(E) in Salmonella-infected macrophages as well as stationary phase cultures. Interactions between alternative sigma factors permit the integration of diverse stress signals to produce coordinated genetic responses. PMID- 15819635 TI - Different proteins bind to the butyrolactone receptor protein ARE sequence located upstream of the regulatory ccaR gene of Streptomyces clavuligerus. AB - Cell-free extracts from Streptomyces clavuligerus, purified by elution from heparin-agarose with an ARE-containing DNA fragment or by salt elution chromatography, bind to a 26 nt ARE sequence, for butyrolactone receptor proteins (ARE(ccaR)). This sequence is [corrected] located upstream of the ccaR gene, encoding [corrected] the activator protein CcaR required for clavulanic acid and cephamycin C biosynthesis. The binding is specific for the ARE sequence as shown by competition with a 34 nt unlabelled probe identical to the ARE sequence. A brp gene, encoding a butyrolactone receptor protein, was cloned from S. clavuligerus. Sixty-one nucleotides upstream of brp another ARE sequence (ARE(brp)) was found, suggesting that Brp autoregulates its expression. Pure recombinant rBrp protein binds specifically to the ARE sequences present upstream of ccaR and brp. A brp deleted mutant, S. clavuligerus Deltabrp::neo1, produced 150-300% clavulanic acid and 120-220% cephamycin C as compared with the parental strain, suggesting that Brp exerts a repressor role in antibiotic biosynthesis. EMSA assays using affinity chromatography extracts from the deletion mutant S. clavuligerus Deltabrp::neo1 lacked a high-mobility band-shift due to Brp but still showed a [corrected] slow-mobility band-shift observed in the wild-type strain. These results indicate that two different proteins bind specifically to the ARE sequence and modulate clavulanic acid and cephamycin C [corrected] biosynthesis by its action on ccaR gene expression. PMID- 15819636 TI - Antibiotic-induced SOS response promotes horizontal dissemination of pathogenicity island-encoded virulence factors in staphylococci. AB - Although mobile genetic elements have a crucial role in spreading pathogenicity determining genes among bacterial populations, environmental and genetic factors involved in the horizontal transfer of these genes are largely unknown. Here we show that SaPIbov1, a Staphylococcus aureus pathogenicity island that belongs to the growing family of these elements that are found in many strains, is induced to excise and replicate after SOS induction of at least three different temperate phages, 80alpha, phi11 and phi147, and is then packaged into phage-like particles and transferred at high frequency. SOS induction by commonly used fluoroquinolone antibiotics, such as ciprofloxacin, also results in replication and high frequency transfer of this element, as well as of SaPI1, the prototypical island of S. aureus, suggesting that such antibiotics may have the unintended consequence of promoting the spread of bacterial virulence factors. Although the strains containing these prophages do not normally contain SaPIs, we have found that RF122-1, the original SaPIbov1-containing clinical isolate, contains a putative second pathogenicity island that is replicated after SOS induction, by antibiotic treatment, of the prophage(s) present in the strain. Although SaPIbov1 is not induced to replicate after SOS induction in this strain, it is transferred by the antibiotic-activated phages. We conclude that SOS induction by therapeutic agents can promote the spread of staphylococcal virulence genes. PMID- 15819637 TI - Polypharmacy in the elderly: a literature review. AB - PURPOSE: To review the body of literature addressing polypharmacy in individuals aged 60 years and older to (a) determine primary care providers' definition of polypharmacy, (b) explore how polypharmacy was assessed in primary care, and (c) seek tested interventions that address polypharmacy. DATA SOURCES: A systematic review of electronic bibliographic databases (e.g. EBSCOHost, InfoTrac, OVID, FirstSearch, and FirstSearch Deluxe) utilizing the search terms "polypharmacy," "polypharmacy and elderly," "polypharmacy and research," and "multiple medications" for the period January 1991 to October 2003 was completed. The search was supplemented with online site searches of relevant journals and review of reference lists of each article. CONCLUSIONS: Results of the literature review revealed that polypharmacy continues to be a significant issue and little research has been conducted regarding the methods primary care providers utilize to assess polypharmacy. Also, there is a gap in the literature regarding the interventions implemented by primary care providers to address polypharmacy. IMPLICATIONS FOR PRACTICE: The following definition of polypharmacy in clinical practice might be more practical: the use of medications that are not clinically indicated. Selecting appropriate limits for numbers of medications may be counterproductive in populations with multiple comorbidities. PMID- 15819638 TI - Primary care needs of patients who have undergone gender reassignment. AB - PURPOSE: The purpose of this article is to inform nurse practitioners (NPs) about the primary care needs of patients who have undergone gender reassignment, either by hormone therapy alone or in conjunction with surgery. DATA SOURCES: Data sources used were mainly from a review of the literature about gender identity disorder and gender reassignment. Information was also gathered from several leading surgeons on gender reassignment surgical procedures and subsequent clinical considerations. CONCLUSIONS: There is very little written on the primary care clinical ramifications of transsexual patients and how clinicians can adapt their approaches to healthcare delivery to accommodate their special situations. IMPLICATIONS FOR PRACTICE: Implications for practice include how an NP can adapt clinical practice approaches to provide for patients who have undergone gender reassignment. Changes that occur in the transsexual process may warrant noncustomary primary healthcare screening and examination. PMID- 15819639 TI - Toward a predictive model of patient satisfaction with nurse practitioner care. AB - PURPOSE: (a) To determine if caring behaviors of nurse practitioners (NPs), gender of NPs, setting (urban or rural), and age, gender, ethnicity, education, and income of patients were predictors of patient satisfaction; (b) to determine which of these characteristics was the best predictor(s) of patient satisfaction; and (c) begin to develop a conceptual model for explaining patient satisfaction with NP care. DATA SOURCES: Responses to the Caring Behaviors Inventory (CBI) and a demographic inquiry by 348 NPs in Louisiana and completion of the Di'Tomasso Willard Patient Satisfaction Questionnaire (DWPSQ) and a demographic inquiry from 817 patients in Louisiana served as data sources. A predictive modeling design explored which variable(s) is the best predictor of patient satisfaction, and multiple regression was used to determine the equation for the best-fitting line and the optimal model for the best predictor(s) of patient satisfaction. CONCLUSIONS: CBI mean scores were high for all NPs. No statistically significant difference was found between male NPs' and female NPs' total mean CBI scores and between urban or rural total mean CBI scores. DWPSQ mean scores and subscale scores indicated high satisfaction with NP care. No statistically significant relationships were found between the NPs' CBI mean scores and the patients' DWPSQ mean scores. There were significant relationships between the DWPSQ subscales, including Wait Time and Patient Management. Stepwise linear regression revealed that patients' age group was a predictor of DWPSQ total mean scores. IMPLICATIONS FOR PRACTICE: NPs need to be aware of developmental differences in all age groups and the differences in perceptions of care. There are many variables to consider when determining patient satisfaction with care, including the patients' sociodemographic and health variables, the healthcare system, and characteristics of the healthcare providers. Awareness of these variables may affect how NPs deliver care and ensure quality care with which the patients are satisfied. PMID- 15819640 TI - An examination of ethnic differences in cigarette smoking intention among female teenagers. AB - PURPOSE: To describe and compare predictors of cigarette smoking intention, as defined by the theory of planned behavior, among African American, Puerto Rican, and non-Hispanic white female teenagers. DATA SOURCES: A convenience sample of 430 English-speaking female teenagers (141 African American, 146 Puerto Rican, and 143 non-Hispanic white teenagers), aged 13-19 years, who were patients at family planning clinics in eastern Pennsylvania. The data were obtained through a self-administered questionnaire. CONCLUSIONS: Regression analyses demonstrated attitude, subjective norm, and perceived behavioral control predicted 70% of the variance in smoking intention in African American, 69% in Puerto Rican, and 88% in non-Hispanic white teenagers. In addition, ethnic group differences were identified. For African Americans, smoking intention was mediated principally by perceived behavioral control, whereas for Puerto Rican and non-Hispanic white teenagers, attitudes were the greatest predictor of intention to smoke. Moreover, non-Hispanic white smokers had a stronger intention to smoke than either the African American or Puerto Rican smokers. IMPLICATIONS FOR PRACTICE: Cigarette smoking is the leading preventable cause of death in the United States, with the majority of smokers initiating the habit in their teen years. Nurse practitioners are in key positions to counsel young women about the dangers of smoking. The identification of cultural-specific predictors of smoking will assist in the development of tailored antismoking messages. PMID- 15819641 TI - Factors influencing women's decisions to self-treat vaginal symptoms. AB - PURPOSE: To review the research on women's self-treatment of vaginal symptoms, describe factors influencing this phenomenon, identify evidence-based interventions, and suggest strategies for nurse practitioners (NPs) to promote safe and effective self-treatment decisions by women. DATA SOURCES: Research articles identified through Medline and CINAHL databases. CONCLUSIONS: The primary factors influencing women's decisions to self-treat vaginal symptoms were personal (attitudes, beliefs, values, knowledge and experience, and emotions) and environmental (culture, social networks and norms, media, and life context). IMPLICATIONS FOR PRACTICE: Women's self-diagnostic skills and decision making for self-treatment can be improved through education and support from NPs. The traditional office visit may not provide an opportunity to teach women appropriate self-care because many women do not access providers for advice or information. Different methods of providing information to large numbers of women through consumer publications need to be developed and evaluated. PMID- 15819643 TI - Practitioner review: cognitive rehabilitation for children with acquired brain injury. AB - BACKGROUND: The need to address acquired cognitive impairments is increasing in child populations seen across a range of settings. However, current clinical practice following brain injury in children does not necessarily incorporate the use of cognitive rehabilitation models or techniques. The aim of this paper is to review the literature in this area. METHODS: All published interventions targeting the cognitive domains of attention, memory and/or executive function that could be identified were reviewed. Different cognitive rehabilitation techniques are briefly described and the clinical and research implications of the findings are discussed. RESULTS: Eleven papers, involving 54 children and adolescents receiving intervention, were identified. This literature describes generalised cognitive rehabilitation programmes as well as more specific strategies targeting focal deficits. CONCLUSIONS: There is an absence of randomised controlled trials and a very limited number of studies using other methodological approaches, providing at this time no conclusive evidence for the efficacy of cognitive rehabilitation for children with acquired brain injury, but a clear need to address a range of methodological difficulties in this field of enquiry. PMID- 15819644 TI - A longitudinal study of adolescent adjustment following family transitions. AB - BACKGROUND: This study examined the impact of family transitions, that is, parental separation, divorce, remarriage and death, upon the lives of Australian children and adolescents in a longitudinal study of temperament and development. METHODS: Using longitudinal and concurrent questionnaire data, outcomes for young people experiencing transitions were compared with those of a random comparison group whose biological parents remained together. RESULTS: No significant group differences were found with regard to behavioural and emotional adjustment concurrently or across time, nor on academic outcomes and social competence. Significant differences between the groups were revealed in measures of parent teen conflict and parent-child attachment. A number of gender differences were found, with female participants displaying both greater adaptive and maladaptive behaviours. Particular dimensions of temperament, as well as the parent's overall rating of their child as easy or difficult, were found to be important predictors of adjustment status for both transitions and comparison groups. CONCLUSIONS: Despite some methodological limitations, this study illustrated the resilience of children experiencing family disruption across childhood and adolescence. PMID- 15819645 TI - Hopelessness and suicidal ideation among adolescents in two cultures. AB - BACKGROUND: This study examines the cross-sectional and longitudinal associations among cognitive variables, depressive symptoms and suicidal ideation in Hong Kong Chinese and Caucasian American adolescents. METHODS: Community adolescents (n = 2,044) ages 14-18 years from Hong Kong and the United States provided information regarding their suicidal ideation, depressive symptoms, and cognitions (self efficacy, cognitive errors and hopelessness), at two surveys, six months apart. RESULTS: Self-efficacy was a weak unique predictor of suicidal ideation in both cultures. Hopelessness was the strongest of cognitive variables in concurrent associations with suicidal ideation in bivariate and multivariate models, in both cultures, and in both boys and girls. Hopelessness continued to offer unique prediction when depressive symptoms were controlled, both concurrently and prospectively. CONCLUSIONS: Our results support hopelessness theories of suicidal ideation and behavior in Hong Kong, and extend the cognitive theory of suicidality to a modernized Asian culture. PMID- 15819646 TI - Genetic influences on early word recognition abilities and disabilities: a study of 7-year-old twins. AB - BACKGROUND: A fundamental issue for child psychology concerns the origins of individual differences in early reading development. METHOD: A measure of word recognition, the Test of Word Reading Efficiency (TOWRE), was administered by telephone to a representative population sample of 3,909 same-sex and opposite sex pairs of 7-year-old twins. Analyses allowing for sex differences in aetiology were used to estimate the extent to which genetic and environmental influences contribute to normal variation in word recognition and word recognition difficulties, defined by scores below the 5th and 10th percentiles of the unselected sample. RESULTS: Both normal variation in word recognition and impaired word recognition abilities were substantially heritable (h2 = .65-.67; h(g)2 = .37-.72). Environmental influences were primarily shared between twins, rather than specific to each individual, and small to moderate in magnitude. There was evidence for qualitative sex differences. Quantitative sex differences were also suggested at the extremes, with genetic influences being more important as a cause of reading difficulties in boys than in girls. CONCLUSIONS: These findings indicate that early individual differences and impairments in word recognition are principally influenced by genetic factors and may involve partly distinct genetic or environmental effects in boys and girls. Crucially, they also provide evidence that reading impairments are linked genetically to the normal distribution. Genetic risk for early impairments in word recognition is continuous rather than discrete. PMID- 15819647 TI - Prevalence of obesity, binge eating, and night eating in a cross-sectional field survey of 6-year-old children and their parents in a German urban population. AB - BACKGROUND: To assess the prevalence of obesity, obesity-related binge eating, non-obesity-related binge eating, and night eating in five- to six-year-old children and to examine the impact of parental eating disturbances. METHODS: When 2020 children attended their obligatory health exam prior to school entry in the city of Aachen, Germany, 1979 parents (97.9%) filled out a questionnaire on their child's eating habits and weight development in a cross-sectional survey. Anthropometric measurements were collected for all children in a standardized form. RESULTS: Episodes of binge eating were found in 2.0% of the children surveyed and night eating in 1.1%. There was a significant relationship between binge eating and obesity but not between night eating and the child's weight. Children's binge eating and night eating were strongly associated with eating disturbances on the part of their mothers (adjusted odds ratios [95% confidence intervals]: 6.1 [2.7-13.5] and 7.8 [2.1-29.4], respectively) and with a non German native language (adjusted odds ratios [95% confidence intervals]: 2.6 [1.2 5.5] and 11.6 [3.5-38.7], respectively). CONCLUSIONS: In concurrence with studies on adulthood, binge eating is linked to obesity already in early childhood. Children of mothers with eating disorders and children of mothers with a non German native language are at increased risk of developing eating disorders themselves. Future studies should focus on obesity and eating disorders in early childhood; prevention programs should seek to target young children at risk as early as possible. PMID- 15819648 TI - Readiness to recover in adolescent anorexia nervosa: prediction of hospital admission. AB - OBJECTIVES: To determine if motivation to change in anorexia nervosa during treatment is a predictor of hospitalisation in adolescent patients. METHOD: The Anorexia Nervosa Stages of Change Questionnaire (ANSOCQ), the Eating Disorders Inventory-2 (EDI-2) and the Beck Depression Inventory (BDI) were administered to a group of 70 anorexia nervosa patients (mean age 15.6 years). They were all receiving treatment at a specialised Eating Disorder Unit and were at different points in the treatment programme. Admission during 6-9 month follow-up was recorded in 63 of these patients who had been admitted to the Unit. The other 7 patients were contacted by phone to determine if they had been hospitalised in another unit during the follow-up period. RESULTS: Patients who needed hospital admission during follow-up had higher mean scores at first evaluation on some of the EDI-2 scales and on the BDI, lower ANSOCQ scores and were more likely to have been outpatients at first evaluation. In the logistic regression analysis a low ANSOCQ score and being an outpatient at first evaluation were shown to be independent predictors of hospitalisation during follow-up. CONCLUSIONS: Low motivation to change, depressive symptomatology and some EDI-2 scales are related to the necessity of hospital admission in adolescent patients with anorexia nervosa. PMID- 15819649 TI - Continuity and change from early childhood to adolescence in autism. AB - BACKGROUND: This longitudinal study of 48 children diagnosed with autism at 2-5 years of age was designed to test the hypothesis that diagnosis would remain stable for most of the sample but that there would be improvements in symptom severity, adaptive behavior, and emotional responsiveness in adolescence. METHODS: A sample of children with autism assessed in both early and middle childhood were observed in late adolescence with the Autism Diagnostic Observation Scale (ADOS) and their parents were administered the Autism Diagnostic Interview-Revised (ADI-R) and the Vineland Adaptive Behavior Scale. RESULTS: All but 2 adolescents (46 of 48) met lifetime criteria for autism according to the ADI-R, and all but 4 adolescents (40 of 44) met criteria for autism spectrum disorder on the ADOS. In contrast to the continuity in diagnosis, parents described improvements in social interactions, repetitive/stereotyped behaviors, adaptive behaviors, and emotional responsiveness to others' distress in adolescence compared to middle childhood. High-functioning adolescents with autism showed more improvement in these domains than low-functioning adolescents with autism. The extent to which the adolescents were observed to be socially engaged with their peers in school in middle childhood predicted adaptive behavior skills even when intelligence level was statistically constrained. CONCLUSIONS: The developmental trajectory of children with autism appears to show both continuity and change. In this sample, most individuals continued to be diagnosed in the autism spectrum but parents reported improvements in adolescence. The results suggest that social involvement with peers improves adaptive behavior skills, and this argues for focusing intervention programs in this area. In addition, it is clear that high-functioning adolescents improve more than low-functioning individuals not only in cognitive abilities but also in social interaction skills. Thus, any early intervention that impacts the cognitive abilities of young children with autism is likely to have a parallel influence on their social skills as they mature into late adolescence and early adulthood. PMID- 15819650 TI - The relationship between vocabulary, grammar, and false belief task performance in children with autistic spectrum disorders and children with moderate learning difficulties. AB - BACKGROUND: The aim of this study was to examine the relationship between language and theory of mind in children with autistic spectrum disorders (ASD) and children with moderate learning difficulties (MLD). Previous studies have found a strong association between language and theory of mind in a range of groups, but mostly have not included measures of both grammar and vocabulary; including these enables us to speculate about the causal direction of the relationship. METHODS: Fifty-eight children with ASD and 118 children with MLD were given standardised assessments of vocabulary and grammar, along with standard theory of mind tasks. RESULTS: The relationship between language and theory of mind was more evident in children with ASD than in those with MLD, and grammar was a particularly strong predictor of theory of mind performance in children with ASD. Children with MLD performed better on false belief (FB) tasks than did children with ASD, and their performance was more predictable across the different theory of mind tasks. CONCLUSIONS: Language, in particular grammar, and theory of mind appear to be more strongly related in children with ASD than in those with MLD. We speculate that this relationship may be causal, with some grammatical understanding being a precursor of theory of mind. The implications of these findings are discussed in relation to possible routes for compensatory strategies for mentalising in children with ASD. PMID- 15819651 TI - Interactions of task and subject variables among continuous performance tests. AB - BACKGROUND: Contemporary models of working memory suggest that target paradigm (TP) and target density (TD) should interact as influences on error rates derived from continuous performance tests (CPTs). The present study evaluated this hypothesis empirically in a typically developing, ethnically diverse sample of children. The extent to which scores based on different combinations of these task parameters showed different patterns of relationship to age, intelligence, and gender was also assessed. METHODS: Four continuous performance tests were derived by combining two target paradigms (AX and repeated letter target stimuli) with two levels of target density (8.3% and 33%). Variations in mean omission (OE) and commission (CE) error rates were examined within and across combinations of TP and TD. In addition, a nested series of structural equation models was utilized to examine patterns of relationship among error rates, age, intelligence, and gender. RESULTS: Target paradigm and target density interacted as influences on error rates. Increasing density resulted in higher OE and CE rates for the AX paradigm. In contrast, the high density condition yielded a decline in OE rates accompanied by a small increase in CEs using the repeated letter CPT. Target paradigms were also distinguishable on the basis of age when using OEs as the performance measure, whereas combinations of age and intelligence distinguished between density levels but not target paradigms using CEs as the dependent measure. CONCLUSIONS: Different combinations of target paradigm and target density appear to yield scores that are conceptually and psychometrically distinguishable. Consequently, developmentally appropriate interpretation of error rates across tasks may require (a) careful analysis of working memory and attentional resources required for successful performance, and (b) normative data bases that are differently stratified with respect to combinations of age and intelligence. PMID- 15819652 TI - Maternal attributions and expressed emotion as predictors of attendance at parent management training. AB - BACKGROUND: The effectiveness of parent management training (PMT) as a treatment for child behaviour problems is reduced by high attrition rates. One difficulty with engaging mothers is that, by definition, PMT is directed at the parent, yet many parents believe the cause of the problem lies within the child. Hence the model of therapy offered contradicts their understanding about the cause and nature of the problem. Moreover, the emotional consequence of holding child responsibility causal attributions is associated with high expressed emotion (EE), a known predictor of poor compliance with therapy in other child psychiatric disorders. METHODS: Seventy-five consecutive referrals of mothers to a PMT programme were recruited. EE was assessed using the Camberwell Family Interview methodology. Spontaneous causal attributions about their child's problem behaviour were collected from the same interview material and independently coded using the Leeds Attributional Coding System. Attendance data at the PMT programme was collected following completion of programmes. RESULTS: Contrary to expectations, mothers who made child-responsibility attributions and were highly critical about their child's behaviour were no more likely than non blaming, low EE parents to drop out prematurely from a course of PMT. However, expressing an understanding of their own role in managing their child's behaviour was predictive of attendance. Two factors were, however, more closely associated with greater attrition: having been offered a clinical diagnosis and being from a lower socio-economic family. CONCLUSIONS: Socio-economic factors should be viewed as barriers to uptake of services and successful strategies for increasing engagement of families are likely to be economical rather than psychological. However, although the causal attributions mothers make about their child's behaviour did not predict whether they were likely to attend PMT, an exploratory analysis found evidence to suggest that successful engagement with PMT begins early in the referral process and that referring clinicians should ensure treatment options are aligned with diagnoses provided. PMID- 15819653 TI - Guest editorial: does age make a difference? PMID- 15819654 TI - Towards rehabilitative handling in caring for patients following stroke: a participatory action research project. AB - AIM OF THE STUDY: This study aimed to facilitate nurses to take ownership of their moving and handling practice. OBJECTIVES: To (i) facilitate nurses in sharing their insights in moving and handling patients following stroke, (ii) enable nurses to identify facilitators of safer moving and handling practice, and (iii) empower nurses in collaboration with physiotherapists to direct changes in their practice. BACKGROUND: Traditional top down moving and handling training has had variable impact on nursing practice. A moving and handling incident in a stroke unit demonstrated that there were professional development needs for staff in the stroke unit in relation to their moving and handling practice in the care of patients following stroke. METHODS: An insider participatory action research approach was utilized. Data were analysed from focus group meetings, brainstorming sessions, observational studies, and from written reflective accounts. FINDINGS: Peer pressure, communication, rehabilitative handling awareness, teamwork between nurses and physiotherapists, equipment and environmental issues were affecting moving and handling practice. Nurses identified that equipment, environment, communication and teamwork strategies would facilitate them in using rehabilitative moving and handling practice. Nurses in collaboration with physiotherapists directed changes in their practice. Participant staff members felt involved and valued, and reported changes in understanding, in their handling practice, and enhanced teamwork. CONCLUSION: Participatory action research creates a supportive environment, where those directly involved in moving and handling patients can investigate and direct changes in their practice. Thus it is a significant vehicle for delivering professional development in moving and handling practice. Relevance to clinical practice. Professional development initiatives in moving and handling practice must incorporate processes that enhance inter-disciplinary teamwork and value and utilize the views and experiences of the staff who move and handle patients. PMID- 15819655 TI - Describing gerontological nursing: an academic exercise or prerequisite for progress? AB - AIMS AND OBJECTIVES: To develop a practitioner-led description of gerontological nursing and articulate the principles which underpin its practice. BACKGROUND: As the population ages expertise in gerontological nursing will become increasingly important. However, a complete and widely accepted delineation of the rationale, philosophy, knowledge base and special skills for gerontological nursing is yet to be developed. METHODS: As one cycle of a larger action research project informed by realistic evaluation, a representative sample of 30 Scottish nurses worked together as a Community of Practice to develop a description of gerontological nursing that would have utility in their practice and in the development of best practice care guidance. Data collection involved group interviews and records of on-line discussions. Data were analysed using content analysis. The emerging description was verified through external consultation with other nursing colleagues and older people. RESULTS: Nursing older people within Scotland was perceived as low status and participants reported that a positive reframing of gerontological nursing was needed. A description and underlying principles were developed by nurses and used as a lens to shape best practice statements. The description embraces a person-centred and enabling approach to evidenced-based gerontological nursing. CONCLUSIONS: Gerontological practice requires an accessible consensus view and description to support its development. Practising nurses developed such a view and its utility has been seen in the development of best practice statements. Relevance to clinical practice. The description and principles of gerontological nursing can be used to guide nursing practice, practice development and research. PMID- 15819656 TI - Factors influencing the decision of older people living in independent units to enter the acute care system. AB - AIMS AND OBJECTIVES: This South Australian-based study explored and described the factors influencing the decision of older people living in the community in independent living units to enter the acute care system. BACKGROUND: Community based older Australians, an increasing population segment, make choices about support needed to optimize health and well-being. This includes when to enter the acute care system. Entering this system has potential risks as well as benefits. The current South Australian Department of Human Services policies of 'keeping older people out' of the acute care system has implications for prevention and early intervention measures and requires an understanding of how and why older people enter the acute care system. METHOD: In-depth interviews were conducted with older people (N = 31) and their families (N = 10), drawn from three South Australian aged care organizations providing independent living unit accommodation, and focus groups (N = 14) were conducted with stakeholders to identify factors influencing the decision of older people living in independent living units to enter the acute care system. FINDINGS: Analysis of the data revealed eight facets influencing this group of older people's decision-making with respect to entering the acute care system; they were: expectations of support in the independent living unit not being met; the presence/absence of safety nets; lack of after-hours support; the desire to remain independent; the general practitioner as pivotal; the influence of others; perceptions of the emergency department; and having access to information. RELEVANCE TO PRACTICE: These facets provide insights into entry processes, links and relationships that form an interface between primary care, community care, the aged care industry and the acute system. Analysis of these insights highlight prevention and early intervention responses that can promote the health and well-being of older people, potential ways to streamline services, as well as gaps in current services. PMID- 15819657 TI - Patient autonomy in physical restraint. AB - Despite initiatives to raise the awareness of patient autonomy among healthcare providers, the use of physical restraints on frail or confused older patients continues to be a common practice in many healthcare settings. This paper examines the relationship between patient autonomy and the use of physical restraints by drawing on the literature contradicting its efficacy and the assumption that its use is necessary to protect the welfare of patients. It argues that the paternalistic use of physical restraints without patient's informed consent is morally unjustified and is an unequivocal violation of their autonomy. The duty to respect individual autonomy should be extended to a duty to respect the autonomy of older people who are being restrained. Only in this way can their human dignity and quality of life be enhanced. PMID- 15819658 TI - Editorial: the involvement of service users in care, services and policy - comments and implications for nursing development. PMID- 15819659 TI - Involvement of older people in care, service and policy planning. AB - In line with other helping professions nursing has moved from traditional views of technical expertise as the prime shaper of health care, to a more equal relationship with service users and patients which respects their expertise, knowledge, goals and choices. This philosophy has underpinned the movement to involve older people in care, policy and planning activities. The literature review presented in this paper highlights the growing literature on this subject with respect to older people and maps the key issues and debates for nursing. PMID- 15819660 TI - Beyond tokenistic involvement of older people in research - a framework for future development and understanding. AB - Research and development work in health and social care are increasingly required to involve users at all stages of the research process yet there is scant empirical evidence to support the justification of this laudable aim. Evidence does suggest that at present efforts to achieve this are primarily tokenistic and that more work is needed, both to examine what user involvement in research activity actually means, and how this can best be supported. This paper sets out to describe developments to support involvement of older people through work at the Royal Bank of Scotland Centre for the Older Person's Agenda and to identify a number of challenges that this has raised for researchers. These challenges have arisen out of a number of assumptions that underpin the process of user involvement. The paper discusses some of the benefits of working in partnership with older people and identifies strategic issues for consideration in order to promote future partnership working. PMID- 15819661 TI - Transfusion medicine illustrated. Visual inspection of blood units, a necessary practice at blood centers. PMID- 15819662 TI - Insights on donor screening for West Nile virus. PMID- 15819663 TI - Human herpesvirus-8: what (not) to do? PMID- 15819664 TI - Homing in on D antigen immunogenicity. PMID- 15819665 TI - The 2003 West Nile virus United States epidemic: the America's Blood Centers experience. AB - BACKGROUND: A detailed assessment of West Nile virus (WNV) yield is needed to evaluate the effectiveness of the WNV nucleic acid amplification technology (NAT) screening implemented in 2003. STUDY DESIGN AND METHODS: WNV NAT screening and donation data were compiled from members of America's Blood Centers, which collect nearly 50 percent of the US blood supply. WNV RNA screening was performed with either the Gen-Probe/Chiron Procleix transcription-mediated amplification assay or the Roche TaqScreen polymerase chain reaction. Results of alternate NAT and WNV immunoglobulin M (IgM) antibody assays conducted on index and follow-up samples were obtained from test manufacturers. Presumed WNV positivity was based on NAT repeat reactivity of the individual index donation whereas confirmatory status was based on additional IgM testing of the index donation and NAT and serology testing of follow-up samples. RESULTS: From July through October 2003, 2.5 million donations were screened for WNV RNA. Of 877 NAT-reactive donations (screening positivity rate of 3.5 per 10,000 units), 430 (49%) were confirmed positive, whereas 68 (8%) lacking follow-up data remained presumed positive. The sensitivity and positive predictive value of a presumed viremic result relative to final confirmatory status were 92 and 99 percent, respectively. WNV activity was highest in the central plains with prevalence per 10,000 peaking August 1 to 15 in Colorado (67.7) and South Dakota (77.5) and August 16 to 31 in Wyoming (74.1) and North Dakota (102.0). CONCLUSIONS: WNV screening interdicted many viremic units, thereby reducing transfusion-transmitted infections. This study demonstrates that a national collaborative effort facilitates timely surveillance of blood donor infectious disease prevalence rates. PMID- 15819666 TI - Detection of West Nile virus RNA and antibody in frozen plasma components from a voluntary market withdrawal during the 2002 peak epidemic. AB - BACKGROUND: The US West Nile virus (WNV) epidemic in the summer and fall of 2002 included the first documented cases of transfusion-transmitted WNV infection. In December 2002, the FDA supported a voluntary market withdrawal by the blood banking community of frozen blood components collected in WNV high-activity areas. At the time, the prevalence of viremia and serologic markers for WNV in the blood supply was undefined. STUDY DESIGN AND METHODS: In collaboration with America's Blood Centers, 1468 frozen plasma components (of approx. 60,000 frozen units voluntarily withdrawn from the market) were selectively retrieved from the peak epidemic regions and season (June 23, 2002-September 28, 2002). These units were unlinked, subaliquoted, and tested by WNV enzyme immunoassays (EIAs; Focus Technologies and Abbott Laboratories) and nucleic acid amplification tests (NATs; Gen-Probe Inc. and Roche Molecular Systems). RESULTS: Of the 1468 EIA results from Abbott and Focus, 7 were anti-immunoglobulin M (IgM)- and anti immunoglobulin G (IgG)-reactive by both assays, 8 and 1 were IgM-only-reactive, and 8 and 23 were IgG-only-reactive, respectively. NAT by Gen-Probe and Roche Molecular Systems yielded one RNA-positive, antibody-negative unit containing approximately 440 RNA copies per mL. An additional 10-fold replicate NAT testing by Gen-Probe on 14 of 15 IgM-reactive specimens yielded 2 additional IgM- and IgG reactive units with low-level viremia (i.e., 7/10 and 2/10 replicates tested reactive). CONCLUSION: The prevalence of acute (RNA-positive) and recent (IgM seroreactive) WNV infections indicates that transfusion risk in high-risk areas could have been considerable and that voluntary market withdrawal of frozen components likely averted some WNV transfusion transmissions. The existence of very-low-level viremic units raises concerns, because WNV minipool NAT screening will miss such units and individual NAT may not completely correct this situation. PMID- 15819667 TI - West Nile virus in Canadian blood donors. AB - BACKGROUND: Blood donation screening for West Nile virus (WNV) RNA by nucleic acid testing (NAT) was implemented in Canada in July 2003, and 14 WNV RNA positive donations were identified. Samples were screened in minipools of six donations with a WNV assay (TaqScreen, Roche). Two of the donors were identified by single-donor screening that was initiated in the province of Saskatchewan, which had the highest prevalence of WNV in the country, in early September 2003. STUDY DESIGN AND METHODS: The original 14 samples and follow-up samples (2-35 days after donation), available from 13 of the 14 donors were tested with an in house, real-time, quantitative WNV NAT assay that was specific for WNV. A Health Canada reference reagent was used for calibration. Immunoglobulin M (IgM) and immunoglobulin G (IgG) levels were determined with commercial enzyme-linked immunosorbent assay kits. RESULTS: All donors tested positive for the presence of WNV with the in-house assay. Two donors, 18 and 19, identified by single-donor testing, had extremely low levels of viremia and that could only be detected in 1:38 or 1:39 replicate tests. The titers of the remaining index samples ranged from below log2.8 (the limit of quantitation) to log4.7 NAT detectable units per mL. Three samples, from Donors 17, 18, and 19, were IgM-positive, whereas samples from Donors 18 and 19 were also IgG-positive. The remaining 10 donors with follow up samples all seroconverted. CONCLUSION: The 14 WNV donor samples detected by routine screening were confirmed as WNV RNA-positive by a WNV RNA-specific in house assay and by demonstration of seroconversion in 13 of the 14 donors. PMID- 15819668 TI - Analytical and clinical sensitivity of West Nile virus RNA screening and supplemental assays available in 2003. AB - BACKGROUND: Transfusion-transmitted West Nile virus (WNV) infections were first reported in 2002, which led to rapid development of investigational nucleic acid amplification tests (NAT). A study was conducted to evaluate sensitivities of WNV screening and supplemental NAT assays first employed in 2003. STUDY DESIGN AND METHODS: Twenty-five member-coded panels were distributed to NAT assay manufacturers. Panels included five pedigreed WNV standards (1, 3, 10, 30, and 100 copies/mL), 15 or 16 donor units with very-low-level viremia identified through 2003 screening, and four or five negative control samples. Samples were tested neat in 10 replicates by all assays; for NAT screening assays, 10 replicates were also performed on dilutions consistent with minipool (MP)-NAT. The viral load distribution for 142 MP-NAT yield donations was characterized, relative to the analytical sensitivity of MP-NAT systems. RESULTS: Analytical sensitivities (50% limits of detection [LoD] based on Poisson model of detection of WNV standards) for screening NAT assays ranged from 3.4 to 29 copies per mL; when diluted consistent with MP pool sizes, the 50 percent LoD of screening NAT assays was reduced to 43 to 309 copies per mL. Analytical sensitivity of supplemental assays ranged from 1.5 to 7.7 copies per mL (50% LoD). Detection of RNA in donor units varied consistent with analytical LoD of assays. Detection of low-level viremia after MP dilutions was particularly compromised for seropositive units, probably reflecting lower viral loads in the postseroconversion phase. Based on the viral load distribution of MP-NAT yield donations (median, 3519 copies/mL; range, < 50-690,000), 13 to 24 percent of units had viral loads below the 50 percent LoD of screening NAT assays run in MP NAT format. CONCLUSION: WNV screening and supplemental assays had generally excellent analytical sensitivity, comparable to human immunodeficiency virus-1 and hepatitis C virus NAT assays. The presence of low-level viremic units during epidemic periods and the impact of MP dilutions on sensitivity, however, suggest the need for further improvements in sensitivity as well as a role for targeted individual-donation NAT in epidemic regions. PMID- 15819669 TI - Possible transmission of human herpesvirus-8 by blood transfusion in a historical United States cohort. AB - BACKGROUND: Transmission of human herpesvirus-8 (HHV-8) by blood transfusion in the United States appears plausible but has not been demonstrated. The objective of this study was to evaluate evidence of HHV-8 transmission via blood transfusion. STUDY DESIGN AND METHODS: Serum specimens were collected before and 6 months after surgery from 406 patients who enrolled in the Frequency of Agents Communicable by Transfusion study (FACTS) in Baltimore, Maryland, from 1986 to 1990. The change in HHV-8 serostatus was measured by a lytic-antigen immunofluorescence assay. RESULTS: Of the 284 patients who were initially HHV-8 seronegative and who received transfusions, 2 seroconverted, 1 with a postsurgery antibody titer of 1:160 and the other with a titer of 1:1280. These patients received 12 and 13 units of blood, respectively. None of the HHV-8-seronegative patients who did not receive transfusions seroconverted. If seroconversion was caused by transfused blood, the transmission risk per transfused component was 0.082 percent. CONCLUSIONS: This is the first report suggesting transmission of HHV-8 via blood components in the United States. Because linked donor specimens were not available, other routes of transmission cannot be excluded; however, the evidence is consistent with infection being caused by transfusion. Future studies should include contemporary US populations with linked donor specimens and populations at higher risk for HHV-8 infection. PMID- 15819670 TI - Variation in concentration of prion protein in the peripheral blood of patients with variant and sporadic Creutzfeldt-Jakob disease detected by dissociation enhanced lanthanide fluoroimmunoassay and flow cytometry. AB - BACKGROUND: A highly sensitive dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) and flow cytometry techniques have previously been developed and employed to characterize soluble cellular prion protein (PrP(c)) expression in whole blood and separated components from healthy adult blood donors. No previous studies with these techniques have evaluated the concentration and expression of PrP in the blood of patients with variant Creutzfeldt-Jakob disease (vCJD). STUDY DESIGN AND METHODS: For blood from vCJD patients, sporadic CJD (sCJD) patients, non-CJD neurological controls, and healthy adults, PrP(c) was measured by DELFIA and cell-associated PrP was measured by flow cytometry. RESULTS: DELFIA analysis identified a significant reduction in the concentration of PrP(c) in the whole blood of vCJD (p = 0.012) and non-CJD neurological patients (p = 0.0004) compared with healthy adults. A significant elevation was found in plasma PrP(c) in sCJD patients compared with healthy adult (p = 0.022) and neurological controls (p = 0.050). Flow cytometry found no significant differences between groups in expression of PrP on platelets and lymphocytes, nor in sensitivity of cellular PrP to proteinase K. Neurological controls show significantly less PrP on red cells than healthy adults. CONCLUSION: There are differences in free and cell associated PrP found in blood of CJD patients and control groups, some of which might be useful with other tests in disease profiling as an aid to diagnoses. PMID- 15819671 TI - Clinical significance of bacteriologic screening in platelet concentrates. AB - BACKGROUND: Despite routine bacterial screening with a bacterial culturing system (BacT/ALERT, bioMerieux) of platelet (PLT) concentrates, two cases of life threatening sepsis attributed to transfused PLT products contaminated with Bacillus cereus were reported to the regional hemovigilance office in the southwest region of the Netherlands. These reports necessitated a retrospective evaluation of the currently applied bacteriologic screening program. STUDY DESIGN AND METHODS: Bacteriologic screening of all PLT concentrates on production was introduced in October 2001. Aliquots (5-10 mL) of pooled PLT concentrates in additive solution were taken for cultures with the BacT/ALERT system 14 to 24 hours after donation. The total culturing period was 7 days and in case of positive signals, identification cultures were taken from the culture bottles. The results of the bacterial screening, identification, and clinical significance of possibly contaminated pooled PLT concentrates were evaluated retrospectively over a 2-year period. RESULTS: In this period, a total of 28,104 pooled PLT concentrates were produced. Positive bacterial screening was found in 0.72 percent (n = 203). Of these, in 184 pooled PLT concentrates bacteria were cultured and identified, and in the remaining 19 (9.4%) identification cultures were negative. Before a positive screening was found, 113 PLT concentrates had already been transfused without the occurrence of clinical significant transfusion reactions. CONCLUSION: Bacterial contamination of pooled PLT concentrates was not related to clinically significant transfusion reactions. Despite negative screening for bacterial contamination, life-threatening transfusion-transmitted infections by contaminated PLT products can still occur. Other strategies should be applied to guarantee a higher degree of bacteriologic safety. PMID- 15819672 TI - Anti-D immunization by DEL red blood cells. AB - BACKGROUND: No data are available on the immunogenicity of extremely weak D variants called DEL. Evaluation of alloanti-D formation in a D- female patient after transfusion of apparently D- blood from an Austrian donor led to discovery of a so far unknown DEL type. STUDY DESIGN AND METHODS: Standard blood group serologic methods were applied. Molecular typing, RHD sequencing, and D epitope mapping was performed and the absolute D antigen density determined. RESULTS: After transfusion of RBCs typed D- by routine serology, the recipient developed alloanti-D. Further evaluation with an indirect antiglobulin test confirmed donor RBCs to be D-. Molecular typing, however, demonstrated the presence of the RHD gene in one donor, and RHD sequencing revealed a deletion of four nucleotides in RHD intron 5 (RHD IVS5-38del4) as the only difference compared to the normal RHD gene. Adsorption-elution techniques demonstrated a DEL phenotype without apparent loss of D epitopes. CONCLUSION: This study documents the clinical significance of the DEL phenotype in blood units that was capable of inducing anti-D in a recipient. Qualitative data are provided on D epitope expression in DEL RBCs. PMID- 15819673 TI - Presence of RHD in serologically D-, C/E+ individuals: a European multicenter study. AB - BACKGROUND: RHD blood group alleles with reduced or absent antigen expression are a clinically significant and heterogeneous group. STUDY DESIGN AND METHODS: To detail population genetics data on apparently D- individuals in central Europe, a six-center study was performed with participants from Austria, Germany, Slovenia, Switzerland, and Russia. A total of 1700 serologically D- samples, positive for C and/or E, were investigated. RESULTS: Observed unexpressed RHD alleles were 59 RHD-CE-D+ hybrid alleles, 9 apparently regular RHD, 1 new RHD(Y401X); DELs were 8 RHD(M295I), 6 RHD(IVS3+1G>A), and 1 new RHD(X418L); and weakly expressed RHDs were 2 weak D type 5, 1 weak D type 1, 1 RHD category VI type 1, and 1 novel weak D type 26. Although weak D type 26 was shown to have one of the lowest D antigen densities ever observed, it gave rise to anti-D immunization in a transfused D- individual. CONCLUSION: The relative occurrence of RHD among serologically D- samples, positive for C and/or E, differed significantly in the investigated central European regions. Considering the growing use of molecular typing techniques, correct identification of blood group alleles with scarce or missing antigen expression is of utmost clinical importance and requires reliable population-based frequency data. PMID- 15819674 TI - Massive antenatal fetomaternal hemorrhage: evidence for long-term survival of fetal red blood cells. AB - BACKGROUND: Massive fetomaternal hemorrhage (FMH) can lead to life-threatening anemia. Quantification based on flow cytometry with anti-hemoglobin F (HbF) is applicable in all cases but underestimation of large fetal bleeds has been reported. A large FMH from an ABO-compatible fetus allows an estimation of the life span of fetal red blood cells (RBCs) in the maternal circulation. CASE REPORT: The mother went to the obstetrician twice antepartum owing to symptoms assumed to be preeclampsia; that, however, was not found. She later delivered by cesarean section owing to diminished fetal movements. No fetal weight gain was observed during the last 2 weeks of pregnancy. STUDY DESIGN AND METHODS: Fetal RBCs were quantified by flow cytometry with anti-HbF, anti-Fy(a), anti-s, and anti-Jk(b) on a regular basis. RESULTS: The infant had anemia at delivery and an FMH was determined to be 314 +/- 17 mL (+/- SD) of whole blood. It is assumed that the two antenatal visits were associated with the FMH. Postpartum follow-up showed that fetal RBCs in the maternal circulation were detectable with anti-HbF up to 119 days. Quantification by flow cytometry based on anti-HbF was in agreement with quantification based on anti-Fy(a), anti-s, and anti-Jk(b), although they were less sensitive. CONCLUSION: ABO-compatible fetal RBCs from an FMH had a life span in the maternal circulation close to that of adult RBCs. PMID- 15819675 TI - Genetic basis of the K(0) phenotype in the Swedish population. AB - BACKGROUND: The absence of all Kell blood group antigens (K(0) phenotype) is very rare. K(0) persons, however, can produce clinically significant anti-Ku (K5) after transfusion and/or pregnancy and require K(0) blood for transfusion. Ten alleles giving rise to the K(0) phenotype have been reported: different populations were studied although none from Scandinavia. STUDY DESIGN AND METHODS: Three K(0) samples were identified by blood banks in Sweden (Uppsala, Umea, and Linkoping) during a 20-year period. Kell antigen typing was performed with standard serologic techniques by the respective blood banks and K(0) status was confirmed by the International Blood Group Reference Laboratory in Bristol, England. Polymerase chain reaction and DNA sequencing of the KEL coding region (exons 1-19) was performed on genomic DNA. RESULTS: The Uppsala K(0) was homozygous for a 1540C>T substitution in exon 13, leading to an immediate stop codon. The Umea K(0) was homozygous for 1023delG in exon 8 that results in a frameshift and a premature stop codon in exon 9. In the Linkoping K(0), a previously reported mutation g>a at +1 of intron 3 was found. CONCLUSION: Two novel and one previously reported null alleles at the KEL locus are described. The identified nonsense mutations abolish expression of the Kell glycoprotein and are thus responsible for the K(0) phenotype in these Swedish families. PMID- 15819676 TI - Fetal-neonatal alloimmune thrombocytopenia and unexpected Glanzmann thrombasthenia carrier: report of two cases. AB - BACKGROUND: Discrepancy between phenotyping and genotyping during the investigation of maternofetal alloimmunization leads to the identification of defects on genes encoding membrane glycoproteins (GPs) IIb and IIIa. STUDY DESIGN AND METHODS: Human platelet (PLT) alloantigen (HPA)-1 and -3 PLT phenotypes and genotypes were performed by use of, respectively, the monoclonal antibody specific immobilization of PLT antigens and the polymerase chain reaction (PCR) sequence-specific priming techniques for up to 2400 families with thrombocytopenic newborn. When a discrepancy was detected, genomic DNA from the mother was amplified for coding sequences of either the GPIIb or GPIIIa genes. The genetic abnormality responsible for the discrepancy was determined by direct sequencing of the PCR products. RESULTS: Two cases of discrepancy were identified among 2400 families tested. In the first case, Mother L, serologically assigned as HPA-1b-homozygous, was genotyped HPA-1a/1b. In the second case, Mother S, serogically defined HPA-3b-homozygous, was genotyped HPA-3a/3b. DNA sequence analysis revealed for Mother L a T(1447)-->C a point mutation within exon 10 of the GPIIIa gene and for Mother S a C(480)-->G point mutation within exon 4 of the GPIIb gene. These mutations reported in Glanzmann thrombasthenia (GT) patients account for nonexpression of the implicated allele on the PLT surface. Thus, the mothers are GT carriers. Alloantibodies, hallmarks of immunization, were not detected in the maternal serum samples. CONCLUSION: Although this situation is rarely encountered, it is important to combine phenotyping and genotyping to avoid false PLT typing assignation and erroneous diagnosis when alloimmunization might occur. PMID- 15819677 TI - The validity of serologic tests for Trypanosoma cruzi and the effectiveness of transfusional screening strategies in a hyperendemic region. AB - BACKGROUND: This study aims at obtaining unbiased estimates of the sensitivity and specificity of existing screening tests for Trypanosoma cruzi and at simulating the effectiveness of alternative screening strategies at different prevalence rates. STUDY DESIGN AND METHODS: A systematic random sample of 400 was taken from 1200 banked serum samples of donors screened between August 1998 and January 1999 in Santa Cruz, Bolivia. Samples were tested with indirect hemagglutination test (IHA), indirect immunofluorescence assay (IFA), and four enzyme-linked immunosorbent assays (ELISAs). Sensitivity and specificity of tests were estimated through latent class analysis. RESULTS: The sensitivity of individual tests ranged from 96.5 to 100 percent, and their specificity from 87.0 to 98.9 percent. Combinations of two tests used in parallel would, even at 40 percent prevalence, only miss approximately 1 infected unit per 10,000 screened. At 5 percent prevalence, however, they would yield 75 to 120 false-positive units per 1000 units screened. Parallel testing with IHA plus ELISA or with IHA plus IFA is marginally more cost-effective, compared to single IHA testing, than single ELISA or single IFA testing, regardless of the T. cruzi prevalence. CONCLUSIONS: Routine blood donor screening for T. cruzi with a single test results in unacceptable numbers of false-negative samples in highly endemic areas or in at risk population groups. Adding a second test seems mandatory, but which one to choose depends on local cost components and feasibility. PMID- 15819678 TI - Development and validation of a mathematical algorithm for quantifying preoperative blood volume by means of the decrease in hematocrit resulting from acute normovolemic hemodilution. AB - BACKGROUND: The interindividual variability of blood volume (BV) is high. The aim of this work was to use the decrease in large-vessel hematocrit (HK(LV)) during acute normovolemic hemodilution (ANH) as an instrument for estimating preoperative BV. STUDY DESIGN AND METHODS: In 39 patients, (Group 1) preoperative ANH was performed. Plasma volume, red cell volume, and HK(LV) were measured before and 30 minutes after ANH, respectively. In a retrospective analysis, a mathematical algorithm was developed for estimating BV before ANH by means of the measured changes in HK(LV) and the amount of blood removed during ANH. To validate the method prospectively, ANH was performed in an additional 10 patients (Group 2). Preoperative BV was estimated and measured in the same way as in Group 1. RESULTS: In Group 1, the mean difference between estimated and measured BV before ANH was 200 +/- 403 mL (5.14 +/- 10.12%; p > 0.05; r = 0.8). In Group 2, the estimated BV before ANH was 41 +/- 348 mL (-0.53 +/- 7.84%) lower than the measured BV (p > 0.05; r = 0.94). In this group, however, two patients with an extraordinarily small (2691 mL) and large (6172 mL) preoperative BV, respectively, were identified correctly by means of the algorithm. CONCLUSIONS: The changes in HK(LV) determined during ANH provide a good "bedside" estimation of preoperative BV. PMID- 15819679 TI - Stored platelets contain residual amounts of tissue factor: evidence from studies on platelet concentrates stored for prolonged periods. AB - BACKGROUND: The advent of new strategies for pathogen reduction has raised the question of whether platelet (PLT) concentrates (PCs) exposed to longer periods of storage retain adequate hemostatic function. STUDY DESIGN AND METHODS: The effects of prolonged storage on adhesive and procoagulant functions of PLTs in PCs have been analyzed. The ability of PLTs to interact with surfaces was assessed by en face electron microscopy. Exposure of anionic phospholipids or tissue factor (TF) antigen on PLTs was assessed by flow cytometry and by immunocytochemical methods at the ultrastructural level. Studies were performed in six different PCs followed 0, 3, 5, 7, and 11 days of storage. RESULTS: A progressive impairment of PLT-adhesive functions was observed after 5 days of storage. A progressive increase in expression of anionic phospholipids and development of procoagulant activity (PCA) measured by a modified prothrombin time (mPT) was observed along the storage. Incubation of PLTs with a specific anti-TF resulted in prolongation of the mPT by approximately 10 to 15 percent. Flow cytometry revealed minimal TF expression at later storage times. Immunocytochemical studies showed minimal TF labeling when studies were performed on PLT whole mounts. Labeling was markedly improved when PLTs were previously exposed to sonication. CONCLUSION: Prolonged storage of PCs was associated with decreased PLT-adhesive capacities and enhanced PCA. Current preparation procedures and storage media have important limitations for preserving PCs for longer than 1 week. PLTs in PCs retain residual amounts of TF as assessed by immunocytochemical and functional assays. The origin and hemostatic significance of this TF should be investigated further. PMID- 15819680 TI - Inactivation of viruses in platelet concentrates by photochemical treatment with amotosalen and long-wavelength ultraviolet light. AB - BACKGROUND: Viral contamination of platelet (PLT) concentrates can result in transfusion-transmitted diseases. A photochemical treatment (PCT) process with amotosalen-HCl and long-wavelength ultraviolet light (UVA), which cross-links nucleic acids, was developed to inactivate viruses and other pathogens in PLT concentrates. STUDY DESIGN AND METHODS: High titers of pathogenic or blood-borne viruses, representing 10 different families, were added to single-donor PLT concentrates containing 3.0 x 10(11) to 6.0 x 10(11) PLTs in approximately 300 mL of 35 percent plasma and 65 percent PLT additive solution (InterSol). After PCT with 150 micromol per L amotosalen and 3 J per cm(2) UVA, residual viral infectivity was assayed by sensitive cell culture or animal systems. RESULTS: Enveloped viruses were uniformly sensitive to inactivation by PCT whereas nonenveloped viruses demonstrated variable inactivation. Log reduction of enveloped viruses for cell-free HIV-1 was >6.2; for cell-associated HIV-1, >6.1; for clinical isolate HIV-1, >3.4; for clinical isolate HIV-2, >2.5; for HBV, >5.5; for HCV, >4.5; for DHBV, >6.2; for BVDV, >6.0; for HTLV-I, 4.2; for HTLV II, 4.6; for CMV, >5.9; for WNV, >5.5; for SARS-HCoV, >5.8; and for vaccinia virus, >4.7. Log reduction of nonenveloped viruses for human adenovirus 5 was >5.2; for parvovirus B19, 3.5->5.0; for bluetongue virus, 5.6-5.9; for feline conjunctivitis virus, 1.7-2.4; and for simian adenovirus 15, 0.7-2.3. CONCLUSION: PCT inactivates a broad spectrum of pathogenic, blood-borne viruses. Inactivation of viruses in PLT concentrates with amotosalen and UVA offers the potential to prospectively prevent the majority of PLT transfusion-associated viral diseases. PMID- 15819681 TI - Efficacy of Epstein-Barr virus removal by leukoreduction of red blood cells. AB - BACKGROUND: Epstein-Barr virus (EBV) infection results in life-long carriage of latent virus in B lymphocytes in the majority of the adult population, including blood donors. The removal of EBV from red blood cell (RBC) components by leukoreduction was assessed. STUDY DESIGN AND METHODS: Sixteen randomly selected fresh AS-5 units were leukoreduced by filtration. B lymphocytes from preleukoreduction specimens and mononuclear cells (MNCs) from postleukoreduction specimens were assayed for EBV DNA with sensitive real-time polymerase chain reaction (PCR). RESULTS: EBV genomes were detected in CD19+ B cells in 14 of 16 preleukoreduced RBC units. EBV genomic copy number in the units ranged from 0.18 to 96.84 per 10(5) B lymphocytes representing approximately 135 to 72,630 total EBV genomes per bag. Leukoreduction rendered all but one unit EBV-negative by PCR. The lone PCR-positive unit after leukoreduction amplified 1.2 EBV genome copies from MNCs recovered from the entire unit of leukoreduced RBCs; this unit had the highest EBV viral load before leukoreduction (72,630 EBV genomes). CONCLUSIONS: These results indicate that a 4-log reduction of EBV genomic copy number can be achieved with leukoreduction of RBC units and renders most RBC units EBV-negative by sensitive PCR. PMID- 15819682 TI - Correlation between in vitro aggregation and thromboxane A2 production in fresh, liquid-preserved, and cryopreserved human platelets: effect of agonists, pH, and plasma and saline resuspension. AB - BACKGROUND: Some of the tests used to assess the quality of fresh and preserved platelets (PLTs) include PLT number, PLT morphology, pH of the PLT medium, PLT response to hypotonic stress, and PLT aggregation to agonists. This study was performed to assess the function of fresh and preserved PLTs by their response to aggregation and their production of thromboxane A2 after in vitro stimulation with agonists. STUDY DESIGN AND METHODS: PLTs isolated by apheresis procedures were stored at 22 degrees C for as long as 5 days and then frozen with 6 percent dimethyl sulfoxide, stored at -80 degrees C, thawed, washed, and resuspended in medium. The effects of agonists and the pH and composition of the medium on PLT aggregation and PLT production of thromboxane A2 after stimulation were measured. RESULTS: The agonists and the pH and composition of the medium affected both the aggregation response and the production of thromboxane A2 by the fresh and preserved PLTs. PLT aggregation response to arachidonic acid (AA) and adenosine diphosphate (ADP) was significantly lower in the cryopreserved PLTs than in the fresh and preserved PLTs. After stimulation with AA and ADP, the cryopreserved PLTs produced more thromboxane than did the fresh and liquid-preserved PLTs. CONCLUSIONS: The agonists and the pH and composition of the medium affected the response to aggregate and produce thromboxane in vitro in both the fresh and the liquid-preserved PLTs. PLT thromboxane A2 production may be a better in vitro test than PLT aggregation to assess PLT function in vivo. PMID- 15819683 TI - A dose-response analysis of lenograstim plus dexamethasone for neutrophil mobilization and collection. AB - BACKGROUND: The objective was to evaluate the dose-response relationship of lenograstim plus dexamethasone for neutrophil mobilization and collection. STUDY DESIGN AND METHODS: In a prospective study, 260 healthy volunteers received oral dexamethasone (8 mg) plus a single subcutaneous injection of glycosylated granulocyte-colony-stimulating factor (G-CSF; lenograstim) at medians of 1.5 (1.0 2.3) microg per kg (n = 43), 3 (2.4-4.1) microg per kg (n = 73), 6 (4.3-7.9) microg per kg (n = 123), and 12 (8.2-17.2) microg per kg (n = 21) and underwent neutrophil collections with a polymorphonucleated neutrophil (PMN) program. White blood cell (WBC) counts and PMN mobilization and collection results were compared, and the severity and clinical significance of donor adverse reactions were evaluated. Fifty-two neutropenic patients (29 children, 23 adults) underwent 271 neutrophil transfusions (GTXs) every other day to maintain WBC levels continuously above 0.5 x 10(9) per L. RESULTS: Within the dose range 1.5, 3, and 6 microg per kg, each doubling step was associated with a 10 to 15 percent PMN increase in peripheral blood up to 32.8 (19.1-49.2) x 10(9) per L (6 microg/kg; p C substitution in three generations of a Chinese family. PMID- 15819692 TI - pH and glucose testing of single-donor apheresis platelets should be discontinued in favor of a more sensitive detection method. PMID- 15819694 TI - Immunology and mathematics: crossing the divide. AB - 'It's high time molecular biology became quantitative, it cries out to a physicist ... for modeling. Modeling isn't a crutch, it's the opposite; it's a way of suggesting experiments to do, to fill gaps in your understanding.' John Maddox, Editor of Nature 1966-73, and 1980-95. PMID- 15819695 TI - Increasing the foreignness of an antigen, by coupling a second and foreign antigen to it, increases the T helper type 2 component of the immune response to the first antigen. AB - It has been proposed that the degree of an antigen's foreignness is important in determining the Th1/Th2 phenotype of the immune response it generates. We test this hypothesis here and partially dissect the underlying mechanism. Immunization of C57BL/6 and hen egg lysozyme (HEL)-transgenic mice, tolerant to HEL at the T cell level, with low doses of sheep red blood cells (SRBC), generated a predominant T helper type 1 (Th1) response in both mouse strains. However, substantial numbers of SRBC-specific Th2 cells were generated when normal, but not HEL-transgenic, mice were immunized with a low dose of the conjugate HEL SRBC. The generation of these anti-SRBC Th2 cells in normal mice required that HEL be coupled to SRBC, since HEL was ineffective in deviating the response to SRBC when present but coupled to another, non-cross-reacting, xenogeneic RBC. This Th2 deviation of the anti-SRBC response by HEL thus requires the operational recognition of HEL epitopes linked to SRBC. Thus increasing the foreignness of an antigen increases its ability to generate Th2 cells. Our findings, in the context of previous observations in related systems, support the proposal that more CD4(+) T-cell/CD4(+) T-cell interactions, mediated by the operational recognition of linked epitopes, are required to generate Th2 cells than Th1 cells. PMID- 15819696 TI - Epicutaneous immunization induces alphabeta T-cell receptor CD4 CD8 double positive non-specific suppressor T cells that inhibit contact sensitivity via transforming growth factor-beta. AB - Since it was previously shown that protein antigens applied epicutaneously in mice induce allergic dermatitis mediated by production of T helper 2 (Th2) cytokines we postulated that this might induce suppression of Th1 immunity. Here we show that epicutaneous immunization of normal mice with a different protein antigen applied on the skin in the form of a patch induces a state of subsequent antigen-non-specific unresponsiveness caused by suppressor T cells (Ts) that inhibit sensitization and elicitation of effector T-cell responses. Suppression is transferable in vivo by alphabeta-T-cell receptor CD4(+) CD8(+) double positive lymphocytes harvested from lymphoid organs of skin patched animals and are not major histocompatibility complex-restricted nor antigen specific. Both CD25(+) and CD25(-) CD4(+) CD8(+) T cells are able to suppress adoptive transfer of Th1 effector cells mediating cutaneous contact sensitivity. In vivo treatment with monoclonal antibodies showed that the cytokines interleukin (IL)-4, IL-10 and transforming growth factor-beta (TGF-beta) are involved in the induction of the Ts cells. Additionally, using IL-10(-/-) mice we found that IL-10 is involved in skin induced tolerance. Further in vitro experiments showed that lymph node cells of skin tolerized mice non-specifically suppress [(3)H]thymidine incorporation by antigen-stimulated immune cells and this effect can be abolished by adding anti-TGF-beta, but not anti-IL-4 nor anti-IL-10 antibodies. These studies indicate the crucial role of TGF-beta in skin induced tolerance due to non-antigen-specific Ts cells and also show that IL-4, IL-10 and TGF-beta play an important role in the induction of epicutaneously induced Ts cell suppression. PMID- 15819693 TI - The intriguing biology of the tumour necrosis factor/tumour necrosis factor receptor superfamily: players, rules and the games. AB - The members of the tumour necrosis factor (TNF)/tumour necrosis factor receptor (TNFR) superfamily are critically involved in the maintenance of homeostasis of the immune system. The biological functions of this system encompass beneficial and protective effects in inflammation and host defence as well as a crucial role in organogenesis. At the same time, members of this superfamily are responsible for host damaging effects in sepsis, cachexia, and autoimmune diseases. This review summarizes recent progress in the immunobiology of the TNF/TNFR superfamily focusing on results obtained from animal studies using gene targeted mice. The different modes of signalling pathways affecting cell proliferation, survival, differentiation, apoptosis, and immune organ development as well as host defence are reviewed. Molecular and cellular mechanisms that demonstrate a therapeutic potential by targeting individual receptors or ligands for the treatment of chronic inflammatory or autoimmune diseases are discussed. PMID- 15819697 TI - The role of macrophages in the in vitro generation of extracellular DNA from apoptotic and necrotic cells. AB - Cell death is a ubiquitous process that occurs by apoptosis or necrosis depending on the triggering event. While apoptotic and necrotic cells differ biochemically, both are cleared by macrophages for elimination. The process is very efficient, although DNA can appear in the blood in various clinical conditions associated with cell death. To define the role of macrophages in the generation of extracellular DNA, in vitro experiments were performed, assessing the release of DNA into the media of apoptotic or necrotic Jurkat cells cultured with RAW264.7 or J774 macrophage cell lines. DNA was measured by a fluorimetric assay using the dye PicoGreen. In these experiments, while necrotic cells alone did not release DNA, in the presence of macrophages, significant amounts of DNA appeared in the medium. This DNA contained sequences from the Jurkat cells and had reduced molecular weight in comparison to cellular DNA. Furthermore, coculture of macrophages with enucleated necrotic Jurkat cells did not release DNA, suggesting that the DNA came from the dead cell. In contrast, Jurkat cells made apoptotic by treatment with either staurosporine or etoposide spontaneously released DNA, while coculture with macrophages caused a decrease in the DNA released. With apoptotic cells, the DNA in the medium showed low molecular weight and laddering whether or not macrophages were present. Together, these results indicate that macrophages play an important role in the generation of extracellular DNA from dead and dying cells, with the effect dependent on how the cell died. PMID- 15819698 TI - Modulated interaction of the ERM protein, moesin, with CD93. AB - CD93 is a cell-surface glycoprotein that has been shown to influence defence collagen-enhanced Fc-receptor or CR1-mediated phagocytosis of suboptimally opsonized targets in vitro, and CD93-deficient mice are defective in the clearance of apoptotic cells in vivo. To investigate the mechanism of CD93 modulation of phagocytic activity, GST fusion proteins containing the 47 amino acid intracellular domain (GST-Cyto), or various mutants of the intracellular domain of CD93, were constructed and used to identify intracellular CD93-binding molecules. The intracellular protein moesin, well characterized for its role in linking transmembrane proteins to the cytoskeleton and in cytoskeletal remodelling, bound to GST-Cyto when either cell lysates or recombinant moesin were used as a source of interacting molecules. An association of moesin with CD93 within intact cells was confirmed by co-capping moesin with CD93 in human monocytes. The moesin-binding site on CD93 mapped to the first four positively charged amino acids in the juxtamembrane region of the CD93 cytoplasmic tail. Interestingly, deletion of the last 11 amino acids from the C terminus of CD93 (GST-Cyto-C11) dramatically increased moesin binding to the cytoplasmic tail of CD93 in the cell lysate assay, but not when the binding of purified recombinant moesin was assessed. Furthermore, moesin binding to CD93 was enhanced by the addition of phosphatidylinositol 4,5-bisphosphate (PIP(2)). Taken together, these data suggest that the interaction of moesin with the CD93 cytoplasmic domain is modulated by binding of other intracellular molecules to the C11 region and implies that a PIP(2) signalling pathway is involved in CD93 function. PMID- 15819699 TI - Cloning and characterization of equine CD89 and identification of the CD89 gene in chimpanzees and rhesus macaques. AB - Immunoglobulin A (IgA) is the major antibody class present in external secretions of mammals. At the vulnerable mucosal surfaces, IgA provides a crucial first-line defence by neutralizing pathogens. Primates also have a substantial level of IgA in serum and although not well understood, the biological role of this IgA depends, at least partly, on its ability to interact with specific receptors (FcalphaRs) on the surface of leucocytes. The human FcalphaR, CD89, was the first IgA Fc receptor to be identified and binding of IgA-coated particles to CD89 triggers numerous cellular effector functions, including phagocytosis, antibody dependent cellular cytotoxicity, and release of inflammatory mediators, all of which may play an important role in both systemic and mucosal immunity. For many years humans were the only species known to express CD89, however, it has recently been cloned from cows and rats. Here, we describe the identification of the CD89 gene in three additional species: horses, chimpanzees, and Rhesus macaques. Equine CD89 was identified at the cDNA level, whereas the chimpanzee and Rhesus macaque genes were identified from the available draft genomic sequence. Interestingly, when compared with humans and other primates, horses, cows and rats have a relatively low concentration of serum IgA, so the role of CD89 in these species is of particular interest. The identification and characterization of CD89 in different species will contribute to a greater understanding of the biological role of IgA and CD89 in mucosal and systemic immunity throughout evolution. PMID- 15819700 TI - Detection of thyroglobulin mRNA as truncated isoform(s) in mouse thymus. AB - Recent studies employing reverse transription-polymerase chain reaction (RT-PCR) have demonstrated the intrathymic presence of mRNA for various autoantigens, including thyroglobulin (Tg). Deliberations on the mechanisms of central tolerance usually assume that this approach detects intact mRNA transcripts that can be translated to express the whole autoantigen in the thymus. In the present study, we tested this assumption using mRNA transcripts of mouse Tg which encode at least 13 pathogenic peptides, scattered over a large (8.5 kb) sequence. We found that mRNA encoding 11 out of these 13 Tg peptides was present in both the thyroid and the thymus of CBA/J mice, with no apparent temporal fluctuations in expression from birth to 12 weeks of age. Interestingly, detection of these sequences was also demonstrable in the liver and kidney, but not in muscle. However, mRNA encoding two pathogenic peptides (amino acids 1-12 and amino acids 1579-1591) was detected intrathyroidally but not in the other tissues. Further analysis by RT-PCR showed that Tg mRNA transcripts in the thymus, liver and kidney lack segments within the 1-915 bp and 961-5013 bp regions, spANNing exons 1-7 and 9-22, respectively. These data strongly suggest that certain known and perhaps other, as yet unmapped, pathogenic T-cell epitopes of Tg cANNot be encoded by the truncated isoform(s) of intrathymic Tg mRNA. These findings also imply that central tolerance to endogenous Tg produced by thymic epithelial cells may be incomplete. PMID- 15819701 TI - Regulation of human neutrophil chemokine receptor expression and function by activation of Toll-like receptors 2 and 4. AB - Neutrophil chemokine receptor expression can be altered by exposure to Toll-like receptor (TLR) agonists, a process that is thought to have the potential to localize neutrophils to sites of infection. In order to investigate this process in more detail, we examined the regulation of highly pure neutrophil CXCR1 and CXCR2 expression and function by selective agonists of TLR2 (Pam(3)CSK(4)) and TLR4 (lipopolysaccharide, LPS). CXCR1 and CXCR2 were down-regulated by TLR engagement. CXCR2 loss was more rapid and showed a dependence upon soluble helper molecules (LPS binding protein and CD14) that was not evident for CXCR1, suggesting differential coupling of LPS signalling to CXCR1 and CXCR2 loss. However, TLR engagement in highly pure neutrophils did not result in complete loss of chemokine receptors, and LPS-treated neutrophils remained able to mount a respiratory burst to CXCL8 and CXCL1, and were able to migrate towards CXCL8 in assays of under-agarose chemotaxis. Thus, although treatment of purified human neutrophils with TLR2 and TLR4 agonists modifies chemokine receptor expression, remaining receptors remain functionally competent. PMID- 15819702 TI - Effect of oestradiol on mouse uterine epithelial cell tumour necrosis factor alpha release is mediated through uterine stromal cells. AB - Oestradiol-17beta (Oe(2)) stimulates uterine epithelial cell proliferation and is critical for normal uterine differentiation and secretory function. Oe(2) can act directly on the epithelium via the epithelial oestrogen receptor (OR) or indirectly via the OR-positive underlying stroma. A primary role for epithelial stromal interactions has been established for mediating steroid hormone action in the uterus. This study was undertaken to determine the mode of Oe(2) action in regulating epithelial cell cytokine release in the uterus. Mouse uterine epithelial and stromal cells were isolated and cultured separately. Transepithelial resistance (TER) was monitored with an EVOM voltohmmeter to determine monolayer polarity and integrity. Epithelial cells grown alone or in coculture with stromal cells were treated with Oe(2). Supernatants collected were assayed for transforming growth factor-beta (TGF-beta) and tumour necrosis factor alpha (TNF-alpha) by bioassay and enzyme-linked immunosorbent assay, respectively. While Oe(2) treatment of epithelial cells led to a significant decrease in TER, the amount of TNF-alpha released was not altered. However, when epithelial cells were cocultured with stromal cells and treated with Oe(2), apical TNF-alpha release was significantly decreased, compared to cells not treated with hormone. As determined by oestrogen receptor antagonist studies, Oe(2) primed epithelial cells for the action of the stromal paracrine factor(s). In contrast, TGF-beta release by epithelial cells was not affected by Oe(2) when grown alone or in the presence of stromal cells. These studies indicate that Oe(2) has both direct and indirect effects on the uterine epithelium. While epithelial monolayer integrity is directly influenced by Oe(2), TNF-alpha release in response to Oe(2) is dependent on the presence of stromal cells, indicating that paracrine communication is necessary for steroid regulation of some but not all cytokines. PMID- 15819703 TI - Neutrophil differentiated HL-60 cells model Mac-1 (CD11b/CD18)-independent neutrophil transepithelial migration. AB - During active intestinal inflammation granulocytes accumulate in the lumen of the gut where they damage the epithelium through the release of various products such as reactive oxygen species and proteolytic enzymes. Previously, using function blocking monoclonal antibodies, we showed that neutrophil migration across intestinal epithelial monolayers in response to various chemoattractants was partially beta(2) integrin Mac-1 (CD11b/CD18)-independent. Here, we show that treating neutrophils with intact monoclonal antibody (mAb) to CD18 activates the cells to express more CD11b. Thus our goal now was to determine whether neutrophil Mac-1-independent transepithelial migration proceeds independently of prior cell activation through Mac-1. We took two approaches, one using blocking Fab' fragments of mAb to CD18 and the second was to develop a neutrophil differentiated HL-60 cell line which is Mac-1 deficient to further study neutrophil/epithelial cell interaction. Anti-CD18 Fab' minimally activated neutrophils but inhibited approximately 75% of transepithelial migration to fMLP while having a minimal effect (70%) and the presence of more than one vascular risk factor in an individual patient were found to be more common in patients with lacunar brain infarcts regardless of aspirin intake. Absence of vascular risk factors was encountered more in IOMP patients. The recurrence of lacunar CVA was significantly higher than recurrence of IOMP. A history of Bell's palsy was more common in IOMP patients than in patients with lacunar CVA. Within the IOMP group, the prevalence of vascular risk factors did not differ between the aspirin and non-aspirin group. Ischemic heart disease (IHD), CVA and recurrence were found more often in the aspirin group. Within the CVA group hypertension, IHD, cardiac arrhythmia and recurrence rate were more common in the aspirin group whereas smoking was found to be more common in the non-aspirin group of patients. CONCLUSIONS: Arteriosclerosis is the main cause of lacunar CVA and IOMP. However, IOMP depends less on the presence of vascular risk factors than does lacunar CVA. Furthermore, aspirin - at least at low doses - does not seem to have a protective effect on either of these conditions, but more extensive prospective studies of homogeneous groups of patients are needed to clarify the preventive role of antiplatelet agents in IOMP. PMID- 15819715 TI - FDG-PET and MRI in potassium channel antibody-associated non-paraneoplastic limbic encephalitis: correlation with clinical course and neuropsychology. AB - OBJECTIVES: We report a patient with potassium channel antibody-associated non paraneoplastic limbic encephalitis (NPLE) in whom repeated fluorodeoxyglucose positron emission tomography (FDG-PET) and magnetic resonance imaging (MRI) are correlated with epileptic activity and memory performance during the course of disease. CASE SUMMARY: A 32-year-old woman suffered from prolonged global amnesia after two generalized tonic-clonic seizures due to NPLE. Initially, MRI showed swelling of the left hippocampus. In FDG-PET, however, bitemporomesial hypermetabolism was seen corresponding to frequent bitemporal independent seizure patterns. Also neuropsychological impairments pointed to a bitemporal involvement at this early stage. In parallel with improved control of electrographic seizure patterns, improvement was seen in FDG-PET and in memory performance. During the whole course, MRI showed only left-sided abnormalities, which correlated with a permanent verbal memory impairment. CONCLUSION: FDG-PET was more sensitive in showing the initial bitemporal involvement and correlated well with EEG findings and neuropsychological impairment in the acute phase of disease. In contrast, structural MRI better reflected persistent neuropsychological deficits. PMID- 15819717 TI - Aberrant methylations in cancer cells: where do they come from? AB - Cancer epigenetics is rapidly moving into a translational phase, and knowledge on how aberrant DNA methylation is induced is becoming important. Aging, chronic inflammation, and viral infections are known to promote methylation of non-core regions of promoter CpG islands (CGI). The non-core methylation and 'seeds of methylation', scattered methylation in a CGI, are considered to serve as triggers for dense methylation of a promoter CGI, which permanently represses expression of its downstream gene. Decreased gene transcription is an important factor that promotes induction of dense methylation. The presence of the CGI methylator phenotype (CIMP), in which methylation of multiple CGI was observed, is under dispute. Some gastric cancer cell lines have increased rates of de novo methylation, and neuroblastoma cases with CIMP show qualitatively different prognosis from those without. This strongly supports the presence of CIMP, but it seems to contain multiple entities. Limited knowledge is available for epimutagens, the chemicals that induce DNA demethylation or methylation. We have developed an assay system to detect demethylating agents, and an assay system for methylating agents is necessary. Efforts in the field on how aberrant methylation is induced will lead to new cancer prevention, diagnostics, and therapeutics. PMID- 15819718 TI - Roles of membrane-type matrix metalloproteinase-1 in tumor invasion and metastasis. AB - Degradation of extracellular matrix (ECM) is one of the first steps in tumor invasion and metastasis. Matrix metalloproteinases (MMP) have been strongly implicated in this step. Membrane-type MMP-1 (MT1-MMP) was first identified as an activator of proMMP-2 expressed on the surface of tumor cells and later, not only ECM macromolecules but also various biologically important molecules, were shown to serve as substrates for MT1-MMP. Accumulated lines of evidence have demonstrated that MT1-MMP expression level is closely associated with invasiveness and malignancy of tumors, suggesting that MT1-MMP is one of the most critical factors for tumor invasion and metastasis. Despite enthusiasm for MMP inhibitors, phase III trials have not yet demonstrated significance in overall survival and side-effects remain an issue. An understanding of the functions of MT1-MMP could supply clues for developing novel therapeutic strategies targeting MT1-MMP. PMID- 15819719 TI - Increase in the frequency of K-ras codon 12 point mutation in colorectal carcinoma in elderly males in Japan: the 1990s compared with the 1960s. AB - The incidence of colorectal carcinomas has been increasing over the last 50 years in Japan. In order to determine whether adenoma-carcinoma sequence (ACS) or de novo cancer development, generally considered to be two separate genetic pathways, might be responsible, K-ras codon 12 mutations, good markers for ACS, were examined in 59 and 84 cases of advanced colorectal cancer surgically resected in Cancer Institute Hospital of the Japanese Foundation for Cancer Research in 1960-1969 and in 1990-1999, respectively. There was no significant difference of K-ras codon 12 mutation between the 25.4% (15/59) in the 1960s and 36.9% (31/84) in the 1990s (P = 0.148), and the reference of distal colon cancer also showed no significant difference between 24.4% (11/45) and 36.4% (20/55). Yet elderly males showed a significant difference: 27.3% (6/22) in the 1960s and 59.3% (16/27) in the 1990s. The references of males, elderly patients (over 75 years old) and distal colon cancer in the 1990s were significantly more likely to demonstrate mutations than their counterparts in the 1960s. There was no variation with the tumor location. The results suggest that the ACS pathway might have primarily contributed to the increased incidence of colorectal cancer in elderly males in Japan. PMID- 15819720 TI - Influence of atrazine administration and reduction of calorie intake on prostate carcinogenesis in probasin/SV40 T antigen transgenic rats. AB - Atrazine, which has been used worldwide as a pesticide, is now known to exert endocrine disrupting (antiandrogenic) effects in mammals. In this study, modifying effects of dietary feeding of 500 and 1000 p.p.m. atrazine on the development of androgen-dependent prostate cancer were investigated using male probasin/SV40 T antigen transgenic (TG) rats. As administration of atrazine has now been identified as causing a decrease in bodyweight, a dietary-restricted TG rat group was also included in order to elucidate the influence of reduction of calorie intake per se on the development of prostate cancer. At week 13, almost the entire lobes of the prostate were occupied with tumor lesions, with no clear intergroup differences in the incidences and multiplicities. Therefore, morphometrical assessment ratios of the prostate epithelial area to the whole prostate tissue area were evaluated. The ratio in the lateral lobe of the 1000 p.p.m. atrazine-treated group was significantly decreased, and there was a tendency to decrease in the ratios in the dorsal lobe of the atrazine-treated groups. However, dietary restriction itself without atrazine treatment caused the same reduction to a similar or greater extent. Testosterone levels were not affected by atrazine administration or dietary restriction. Our results indicate that the observed atrazine-related suppression of prostate carcinogenesis was probably caused by the decrease in calorie intake, rather than by atrazine related endocrine disruption. PMID- 15819721 TI - Expression of RUNX3 protein in human lung adenocarcinoma: implications for tumor progression and prognosis. AB - Runt-related transcription factor 3 belongs to the runt domain family of transcription factors that play a pivotal role during normal tissue development and tumorigenesis in several organs. We directed our attention to the expression of RUNX3 protein in human lung AC and non-neoplastic lung tissues, comparing the results with clinicopathological profiles. We evaluated the expression of RUNX3 protein in 17 pairs of lung AC and non-neoplastic lung tissue. Furthermore, 98 lung AC were studied to examine the frequency of RUNX3-positive cells. Western blot analysis showed a single band at 45 kDa in all 17 AC and non-neoplastic tissues. Immunohistochemistry revealed immunoreactivity in alveolar type II pneumocytes or Clara cells. RUNX3 was expressed more frequently in the carcinomas with a BAC component than in those without (P < 0.01). Lower RUNX3 levels were associated with poorly differentiated types (P = 0.049). The five-year survival rate was significantly higher in the 50 patients with higher levels of RUNX3 expression than in the 48 patients with lower levels (P = 0.027). The expression of RUNX3 protein in lung AC might play a pivotal role in tumor progression and patients' survival. PMID- 15819722 TI - StAR and progesterone producing enzymes (3beta-hydroxysteroid dehydrogenase and cholesterol side-chain cleavage cytochromes P450) in human epithelial ovarian carcinoma: immunohistochemical and real-time PCR studies. AB - Steroidogenic acute regulatory protein (StAR), P450 side-chain cleavage enzyme (P450scc) and 3 beta-hydroxysteroid dehydrogenase enzyme (3beta-HSD) are all involved in the transport of cholesterol and production of progesterone. In situ production of sex steroids including progesterone have been considered to play important roles in pathogenesis and/or development of common epithelial ovarian carcinomas. In this study, StAR, P450scc, and 3beta-HSD were immunolocalized in 100 cases of ovarian carcinoma and results were then correlated with clinicopathological and prognostic parameters of individual patients including status of progesterone receptor (PR) in tumor cells. Results of immunohistochemistry were further characterized by real-time PCR analysis in 20 cases of epithelial ovarian carcinomas in which frozen tissues were available for examination. StAR, P450scc, and 3beta-HSD immunoreactivity was detected predominately in the cytoplasm of carcinoma cells. Results of real-time PCR analysis were correlated with those of immunohistochemical studies. StAR, P450scc, and 3beta-HSD H scores demonstrated significant inversed statistical correlation with FIGO stage, residual size of the tumor, and Ki67 LI. A positive statistically significant correlation was detected between StAR H score and PR-B LI. Multivariate statistical analysis demonstrated that the status of intratumoral StAR, FIGO stage, and residual tumor size all turned out to be independent prognostic factors for the clinical outcome of the patient. The presence of StAR, a cholesterol transporter for steroidogenesis in human epithelial ovarian carcinoma, may reflect the ability of these tumors to produce progesterone in situ that could influence biological behavior of these tumors, especially through progesterone dependent inhibition of tumor cell proliferation. PMID- 15819723 TI - N-acetyl-cysteine enhances growth in BCR-ABL-transformed cells. AB - N-acetyl-cysteine (NAC) has been reported to have anticancer properties such as counteractions against mutagens and prevention of tumor progression by scavenging reactive oxygen species (ROS). However, here we report that NAC can enhance the anchorage-independent growth of cells transformed by activated ABL tyrosine kinases or Ras. This effect was not dependent on loss of focal adhesion kinase activation. NAC rescued cell growth that was suppressed by heat shock protein (Hsp) 90 inhibitors possibly by chemical modification of their quinone moiety. NAC rendered Rat1/BCR-ABL cells resistance to a Ras inhibitor manumycin in soft agar colony formation. In the absence of Hsp90 inhibitors, NAC stimulated the activation of MAP kinase in BCR-ABL-transformed but not in the parental Rat1 cells. We propose that NAC should be used carefully in cancer treatment. PMID- 15819724 TI - Cancer and epigenetics--basic research and clinical implications: Joint Meeting of the Fourteenth International Symposium of the Hiroshima Cancer Seminar and the Eighth Meeting of the Tottori Bioscience Promotion Foundation, October 2004. PMID- 15819725 TI - Psychiatric research in low- and middle-income countries: the need for concrete action. PMID- 15819726 TI - The effectiveness of psychological treatments for treatment-resistant depression: a systematic review. AB - OBJECTIVE: A systematic review of all studies (controlled and uncontrolled) to evaluate psychological interventions with treatment-resistant depression. METHOD: A systematic search to identify studies evaluating a psychological intervention with adults with a diagnosis of major depressive disorder who had not responded to at least one course of antidepressant medication. RESULTS: Twelve studies met inclusion criteria, of which four were controlled and eight uncontrolled. Treatment effect sizes were computable for four studies and ranged from 1.23 to 3.10 with a number of better quality studies demonstrating some improvements in patients following a psychological intervention. CONCLUSION: Psychological treatments for depression are commonly delivered and often recommended following the failure of medication. The paucity of evidence for their effectiveness in these situations is a significant problem. There is a need for studies with a strong controlled design investigating the effectiveness of psychological treatments for patients with treatment-resistant depression. PMID- 15819727 TI - Common mental disorders and use of general health services: a review of the literature on population-based studies. AB - OBJECTIVE: To review population-based studies on the association between common mental disorders and the use of general (non-mental) health care services. METHOD: Literature search in Medline and PsychLit databases. Only studies with a prospective design and correction for somatic morbidity were included for review. RESULTS: On the most general level of outcomes considered and in the majority of studies, mental disorders were associated with higher service use. This general tendency is not consistently reflected in the use of specific health care services, but is materialized in different patterns of out-patient and in-patient service utilization, which vary from study to study. Findings for the elderly were less clear-cut than for other age groups. CONCLUSION: Mental disorders are related to higher general health care service use on a global, aggregated level. These associations are not specific for certain types of services. PMID- 15819728 TI - Mental health needs, service use and costs among Somali refugees in the UK. AB - OBJECTIVE: To assess the mental health needs and service use of Somali refugees living in London. METHOD: Subjects (n = 143) were sampled from conventional and non-conventional sites. Needs and service use were measured using the Camberwell Assessment of Need and the Client Service Receipt Inventory, respectively. Comparisons between sites were made and cost predictors identified. RESULTS: Basic needs occurred frequently but were often not fully addressed. The mean number of needs was around four out of a possible 22. The most used services were GPs, other clinicians and refugee services. Higher non-inpatient costs were associated with length of stay in the UK and lower costs with being at risk of suicide and having panic disorder or agoraphobia. CONCLUSION: Somali refugees living in London have a relatively high level of need but a low level of service use. Refugee characteristics could only account for a limited amount of cost variation. PMID- 15819729 TI - Are immigrants at a disadvantage in psychiatric in-patient care? AB - OBJECTIVE: To assess the utilization of psychiatric in-patient care among immigrants, and to compare immigrants and natives with respect to sociodemographic and clinical characteristics. METHOD: Analysing a sample of 23 377 consecutive referrals to psychiatric hospitals of a catchment area in 1995 2001. RESULTS: Within this sample, 20% were foreign nationals. Rates of psychotic disorders were similar in immigrants and natives. Regarding other diagnoses, sociodemographic and clinical measures, there were significant differences. Most immigrant groups had higher rates of compulsory admission, were more likely to be admitted with lower illness severity and not to be readmitted, and spent significantly shorter time in hospital, compared with Swiss in-patients. Some of these differences were clearly gender-specific. CONCLUSION: Service utilization and psychiatric treatment decisions are not explained merely by illness-related aspects in immigrants. Social and cultural factors have to be recognized in order to prevent disadvantages in psychiatric care. PMID- 15819730 TI - Early behavioral symptoms and course of Alzheimer's disease. AB - OBJECTIVE: To determine if behavioral symptoms detected at initial evaluation relate to cognitive or functional status or survival time in Alzheimer's disease (AD) patients. method: Review, in 100 cases of autopsy-proven AD, of the relationship of behavioral symptoms detected at initial evaluation to cognitive and global function measures and survival time. RESULTS: Behavioral symptoms had occurred in 74% of patients, including apathy (51%), hallucinations (25%), delusions (20%) and depressed mood (6.6%). Verbal aggression was common (36.8%); physical aggression less so (17%). The symptomatic group was more functionally (but not cognitively) impaired and had shorter median survival time (8 years: 95% CI: 7-9 years vs. 10 years: 95% CI: 8-12 years; P = 0.002) than the asymptomatic group. The presence of any one symptom at initial evaluation accounted for 6.1% of the variance in duration of illness. CONCLUSION: Presence of behavioral symptoms at initial evaluation of AD patients is associated with greater functional impairment and shorter survival time. PMID- 15819731 TI - Aversive tension in patients with borderline personality disorder: a computer based controlled field study. AB - OBJECTIVE: This study was designed to examine the subjective appraisal of aversive tension under conditions of daily life in patients with borderline personality disorder (BPD). METHOD: A sample of 63 female subjects with BPD and 40 mentally healthy controls were each given a hand-held computer. For two consecutive days, participants were prompted at hourly intervals to record their current state of aversive tension and prompting events. RESULTS: Compared with controls, states of aversive tension occurred significantly more frequently in BPD patients. The average levels of aversive tension were significantly higher, and the rate of increase in tension was markedly more rapid. Furthermore, states of aversive tension persisted for a longer period. Among BPD subjects three events (rejection, being alone, and failure) account for 39% of all events preceding states of tension. CONCLUSION: The study provides support for the theory that patients with BPD experience more frequent, stronger, and longer lasting states of aversive tension. PMID- 15819732 TI - Assaultive trauma and illness course in psychotic bipolar disorder: findings from the Suffolk county mental health project. AB - OBJECTIVE: Little is known about the relationship of assaultive trauma to clinical and functional outcome in patients with bipolar disorder. METHOD: We assessed trauma histories in a cohort of 109 first-admission bipolar patients with psychosis using structured interviews and medical records. Assaultive trauma included rape, physical attacks, and physical threats. Outcome was assessed using standardized ratings. RESULTS: Forty percent reported a history of assaultive trauma, mostly in childhood (< or =16 years). Exposed patients were more symptomatic at each follow-up than unexposed. Sixteen percent of exposed patients remitted after one episode compared with 38.5% of the non-exposed. Patients exposed as adults were the most symptomatic at 6 months, while patients exposed in childhood were the most symptomatic at 24 months. CONCLUSION: Our findings supported the salient role of trauma history as a risk factor for poor course in severe bipolar disorder. Given the high prevalence of such exposure, clinical awareness in first-admission psychotic bipolar patients is critical. PMID- 15819733 TI - Suicide in classical mythology: cues for prevention. AB - OBJECTIVE: To compare well established antecedents and correlates of completed suicide with the motives and the mechanics reported in Greek mythology. METHOD: A well-known collection of Greek myths, the Book of fables by Hyginus, was explored to investigate the mechanics driving an individual to imagine, design and carry out a suicide attempt. RESULTS: Females outnumber males in the mythographer's list, their favourite methods to die being drowning, hanging, self-burning and throwing themselves down from on high. Some kind of familial recurrence of suicide was accounted for, and a large percentage of these suicides was connected to incest. Shame, sense of guilt and grief for the death of a loved one are the most frequently reported psychological correlates of the act, whereas defeat, failure or a catastrophic change in living conditions and, among females, an unfortunate love affair figure as the main antecedents of suicide. CONCLUSION: Negative life events and emotional reactions to the severing of social ties frequently occur as antecedents of suicide in Greek mythology. PMID- 15819734 TI - CADASIL: underdiagnosed in psychiatric patients? AB - OBJECTIVE: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is exclusively related to symptoms of the central nervous system. Retrospectively in up to 15% the initial presentation is psychiatric disturbances. In these cases the diagnosis often is delayed or missed. METHOD: Two cases of CADASIL diagnosed in a psychiatric hospital are presented. RESULTS: Both patients were admitted to the gerontopsychiatric department (one because of a suicidal attempt and a depressive episode, the other because of cognitive decline and progressive personal neglect). Brain magnetic resonance imaging (MRI) showed severe leukoencephalopathy in the absence of cardiovascular risk factors. In both cases, diagnosis of CADASIL was made by the identification of specific granular osmiophilic material in skin biopsies. CONCLUSION: Brain MRI should be performed in all cases of late onset of severe psychiatric symptoms. CADASIL should be considered as a possible differential diagnosis whenever a marked leukoencephalopathy is detectable. Diagnosis can be verified by taking a skin biopsy or by specific genetic testing. PMID- 15819740 TI - The Israeli Long-Term Care Insurance Law: selected issues in providing home care services to the frail elderly. AB - The paper describes and analyses selected issues related to the provision of home care services to frail elderly people following the Israeli Long-Term Care Insurance Law (1988). The goals and principles of the Law, which mandates the provision of home care services to frail elderly people, are presented. The paper also evaluates its contribution toward enhancing the well-being of elderly clients. Several major dilemmas that arose following implementation of the Law are analysed and evaluated in comparison with other countries that have enacted and implemented similar laws. These dilemmas are community vs institutional care; services in kind vs monetary allowances; service provision through contracting out with nongovernmental agencies; unstable and unskilled labour force; and service quality. Finally, policy implications are discussed, mainly in the following areas: investment in human resources as a condition for achieving high service quality, and the need for coordination between the agencies that provide long-term care services to elderly people. PMID- 15819741 TI - Carers and the digital divide: factors affecting Internet use among carers in the UK. AB - This paper presents data from a cross-sectional survey of 3014 adult carers, examining use of the Internet and factors associated with it. Carers recruited from the databases of three local authorities and other carer organisations within their geographical boundaries and that of Carers UK, a national carers organisation, were sent a postal questionnaire (response rate: 40%). A comparison of our data with national data on carers suggests some under-representation of men and younger adult carers and some over-representation of those who had been caring for long periods and those with substantial caring responsibilities. Two measures of Internet use were used and are presented in this analysis: previous use (ever used vs never previously used) and frequency (less than once a week vs once a week or more). Bivariate analyses identified patterns of Internet use and socio-demographic and socio-economic factors and caring circumstances associated with them. Factors significantly associated with each measure of Internet use were entered into direct logistic regression analyses to identify factors significantly associated with each measure. Half (50%) of all carers had previously used the Internet. Of this group, 61% had used it once a week or more frequently. Factors significantly associated with having previously used the Internet were carer's age, employment status, housing tenure and number of hours per week they spent caring. Frequency of Internet use was significantly associated with carer's age, sex, employment status and number of hours spent caring. Our study suggests that a significant number of carers may not currently be Internet users and that age, gender, socio-economic status and caring responsibilities shape Internet use in particular ways. Given the targets set by government for the development of online services, it is important to address the digital divide among carers and to continue to develop other services and information systems to meet the needs of those who do not access the Internet. PMID- 15819742 TI - Gatekeepers in sickness insurance: a systematic review of the literature on practices of social insurance officers. AB - Decisions concerning entitlement to sickness benefits have a substantial impact on the lives of individuals and on society. In most countries, such decisions are made by staff of private or public insurance organisations. The work performed by these professionals is debated, hence more knowledge is needed on this subject. The aim of the present study was to review scientific studies of the practices of social insurance officers (SIOs) published in English, Danish, Norwegian and Swedish. Studies were searched for in literature databases, in reference lists, and through personal contacts. Analyses were made of type of study, areas investigated, research questions, theories used, and the results. Sixteen studies were included. SIOs and several other actors are responsible for applying measures to minimise sick-leave and promote return to work (RTW). The studies focusing on coordination of such measures revealed that SIOs felt unsure about how to handle their contacts with clients and other actors. One study indicated that the SIOs, partly due to lack of time, accepted the recommendations of physicians instead of making their own judgments about granting sickness benefits. While all SIOs must make decisions concerning entitlement to sickness benefits on a daily basis, few of the reviewed studies scrutinised the actual granting of sickness compensation. The studies were also deficient in that they investigated the decision latitude of the SIOs from a very limited perspective, mainly on an individual level and often primarily in relation to colleagues and/or clients rather than to the laws and regulations of the sickness insurance. The concepts and framework in this area of research need to be developed to facilitate elucidation of the interaction between different actors in local spheres, professionals in different disciplines, and between welfare staff and individual citizens. PMID- 15819743 TI - The self-reported health condition of women after their participation in a stress management programme: a pilot study. AB - From a public health perspective, it is important to develop effective measures to deal with stress which are based on the individual's participation, such as stress management provided in group sessions. Therefore, the aim of the present study was to compare and evaluate the self-reported health condition of women in terms of their general symptoms, stress and sense of coherence (SOC) after participation in a stress management programme. The intervention, which had a modified crossover design and involved 40 women divided into two groups (G1 and G2), comprised eight meetings, the content of which was both theoretical and practical, and included information about stress, stress management, massage and mental training. A questionnaire was filled in on three occasions: before and after the intervention (8 weeks later), and after another 8 weeks (16 weeks later). The questionnaire contained 60 items comprising background factors, general symptoms, stress and SOC. No significant differences existed between the groups at baseline. In favour of the intervention, significant differences were found between the groups with regard to fewer general symptoms (P = 0.035) as well as a tendency to stress reduction (P = 0.060). A comparison within groups showed that both groups had a significant reduction in symptoms after the intervention (G1, P = 0.002; and G2, P = 0.003) and in reduced stress (both P = 0.001). After a further 8 weeks, both groups still showed significantly fewer general symptoms and reduced stress, as well as significant improvements with regard to SOC (G1, P = 0.012; and G2, P = 0.026). These findings indicate that the combination of mental training and massage in this stress management programme had a positive influence on the women's health condition. The pilot study design could be used in a full-scale study with randomised groups. PMID- 15819744 TI - Support for very old people in Sweden and Canada: the pitfalls of cross-cultural studies; same words, different concepts? AB - The Swedish and Canadian 80+ studies are collecting longitudinal data regarding medical, psychological and sociological aspects of the lives of people who are 80 years of age and older. A paper entitled 'Informal and formal support from a multidisciplinary perspective: a Swedish follow-up between 80 and 82 years of age', was based on the Swedish 80+ study. It examined psychosocial and health measures by support type (i.e. no support, and formal and informal support). Support is defined as care given by either friends and family (informal support) or healthcare professionals (formal support). The present paper compares participant characteristics between two groups of 80-year-olds from two study sites (i.e. Lund, Sweden, and Fredericton, NB, Canada), using the Swedish study site data. Only those participants who participated at both 80 and 82 years of age were included in the analyses. The intent of the descriptive analyses was to compare cross-culturally the two groups of participants and the level of support that they received. Between the two groups, the level of support received by the participants was rather dissimilar: Canadians reported receiving far more informal and formal support compared to Swedes. This finding was despite the fact that the Swedish state provides more funded support than that of Canada to its citizens. Therefore, the present authors speculate that the concept of support has a different meaning in Sweden than in Canada. This speculation raises concerns about cross-cultural studies, particularly when survey questions involve culturally ambiguous concepts such as the term 'support'. PMID- 15819745 TI - The mental health of looked after children: matching response to need. AB - Previous research has established high rates of mental health need in looked after children. The study described in this paper aimed to explore in more depth the mental health problems of looked after children and to examine the service response to those needs in two English local authorities. The study utilised a purposive sample of 80 looked after children which was biased towards those with high needs but which reflected the characteristics of looked after children in the two areas with regard to age, gender and type of placement. Key data on children and young people were collected from social services' case files and a set of indicators of mental health need was designed by the research team in order to distinguish different types and levels of mental health problems. High levels of mental health need in the study group were associated with placement disruption. Educational difficulties were also apparent in the group with the highest levels of mental health need. Longer-term input from child and adolescent mental health services (CAMHS) professionals did not appear to be targeted on the group with the highest level of mental health needs. The researchers conclude that longer-term CAMHS interventions could be usefully focused on looked after children who have experienced high levels of placement disruption. As the professional group most likely to be providing substantial intervention to looked after children and their carers, social workers require relevant training in identifying and responding to mental health needs. CAMHS professionals could develop and strengthen their consultative roles with front-line carers, social workers and schools. PMID- 15819746 TI - Providing welfare advice in general practice: referrals, issues and outcomes. AB - General practices in the UK are increasingly hosting welfare advice services on their premises to address patients' social and economic needs. In this paper, the authors present the outcomes of a service providing welfare advice across 30 general practices in inner-city Bradford. A retrospective study of all patients referred for advice during the initial 24 months of the project was conducted. The following information was collected: patient demographics, source of referrals, advice issues raised and income generated through benefit claims. The advice workers saw 2484 patients dealing with over 4000 welfare advice issues. Demand for the service varied widely across practices, reflecting practice list size and engagement with the service by practice staff. The main source of referrals was general practitioners (28%), and disability-related welfare benefits constituted the largest category of advice issues. Sixty-nine per cent of patients seen for advice were of south Asian ethnic origin. The advice workers raised pound sterling 2,389,255 in welfare benefit claims for patients, primarily through disability-related benefits. Approximately one in four patients referred for advice benefited financially. It is concluded that the service is an excellent strategy by which primary care organisations address the social, economic and environmental influences on the health of their population. PMID- 15819747 TI - Keeping warm and staying well: findings from the qualitative arm of the Warm Homes Project. AB - This paper presents findings from the qualitative arm of the Warm Homes Project, a programme of research concerned with the nature of fuel poverty, its alleviation and its relationship to family health. Much of the research into fuel poverty, which results from various combinations of low income and fuel inefficiency, has drawn upon quantitative paradigms. Experiences of, and coping with, fuel poverty have not been well explored. Data for the present study were obtained through qualitative interviews with household members about the above issues. The findings suggest that the expectations of those in fuel poverty about staying warm, and their beliefs about the relationship between warmth and health, vary considerably. Fuel poverty often had wider ramifications, impacting on quality of life in complex ways. The respondents took steps to alleviate cold, but their strategies varied. Coping was affected by informational limitations as well as cost constraints. Measures designed to alleviate fuel poverty should take into account its wider social meaning within the lives of household members. PMID- 15819748 TI - Involving older people in research: methodological issues. AB - The policy imperative to increase public participation in health and social care research, planning and service delivery raises significant questions about optimum approaches, methods and the extent to which this policy can influence change in practice. This paper highlights the key policy literature on user involvement and participatory research methods to establish the context for a partnership research project exploring perceptions of risk in relation to falls from the perspectives of older people, carers, and health and social care professionals. The paper reports the methods used in developing user involvement in the research at a number of levels, including project management, and a consumer panel working alongside the research team and influencing the dissemination in local falls prevention strategies. The authors draw out the issues related to the context and method of involvement, and discuss the impact on research quality and local service development in health and social care. PMID- 15819750 TI - A cross-sectional behavioral genetic analysis of task persistence in the transition to middle childhood. AB - Task persistence, measured by a composite score of independent teacher, tester and observer reports, was examined using behavioral genetic analysis. Participants included 92 monozygotic and 137 same-sex dizygotic twin pairs in Kindergarten or 1st grade (4.3 to 7.9 years old). Task persistence was widely distributed, higher among older children, positively associated with standardized tests of cognitive performance and achievement, and negatively associated with parents', teachers' and observers' reports of behavioral problems. Cross sectional analysis indicated a strong developmental shift from shared environment variance among younger children to additive genetic variance in older children. PMID- 15819751 TI - Young children have difficulty ascribing true beliefs. AB - Using the format of a false belief task (Wimmer & Perner, 1983), we investigated the ability of 88 3- and 4-year-olds to ascribe a previously held true belief to a story protagonist. In an unexpected transfer task, children found true belief ascription as difficult as false belief ascription even though they could answer memory questions about story details. Results are discussed in relation to theoretical accounts of theory of mind development that stress the importance of understanding the falsity of belief, and those accounts that stress the importance of information or executive processes. PMID- 15819752 TI - Doublet challenge: form comes before function in children's understanding of their orthography. AB - Several current spelling models suggest that children cannot have any knowledge of orthographic form before they have acquired knowledge about orthographic function. We evaluated this proposition by using an orthographic choice task to inspect Finnish schoolchildren's knowledge of two aspects of consonant doublet use: the allowed doublet position (an aspect of orthographic form) and the type of phonemic information they represent (an aspect of orthographic function). The results challenged the view of the existing spelling models, since they showed that already at the beginning of the first school year children possessed formal knowledge of doublet use and knew that word-initial doublets are not allowed. However, these children were ignorant of the function of doublets, i.e. that they stand for long consonants. PMID- 15819753 TI - Processing demands in belief-desire reasoning: inhibition or general difficulty? AB - Most 4-year-olds can predict the behavior of a person who wants an object but is mistaken about its location. More difficult is predicting behavior when the person is mistaken about location and wants to avoid the object. We tested between two explanations for children's difficulties with avoidance false belief: the Selection Processing model of inhibitory processing and a General Difficulty account. Children were presented with a false belief task and a control task, in which belief attribution was as difficult as in the false belief task. Predicting behavior in light of the character's desire to avoid the object added more difficulty in the false belief task. This finding is consistent with the Selection Processing model, but not with the General Difficulty account. PMID- 15819754 TI - Breakdown of spatial parallel coding in children's drawing. AB - When drawing real scenes or copying simple geometric figures young children are highly sensitive to parallel cues and use them effectively. However, this sensitivity can break down in surprisingly simple tasks such as copying a single line where robust directional errors occur despite the presence of parallel cues. Before we can conclude that this directional bias is a limiting case it needs to be shown that any given parallel cues are indeed being processed. We achieved this by using a texture-defined contour where parallel cues are integral to the stimulus and thus cannot be ignored. Here we report that even when prolific parallel cues must have been processed, young children make robust directional errors when copying single lines. This finding demonstrates a major constraint on young children's spatial parallel coding. PMID- 15819755 TI - Early development of delinquency within divorced families: evaluating a randomized preventive intervention trial. AB - This paper reports on an experimental test of coercion theory early onset model of delinquency. Results are from the Oregon Divorce Study-II, a randomized preventive intervention trial with a sample of 238 recently separated mothers and their sons in early elementary school. The objective was to experimentally manipulate parenting variables hypothesized to influence development of delinquent behaviors. Multiple-method assessment spanned 36 months. Because the intervention focused on parent training, we expected that any intervention effects on changes in child outcomes would be mediated by hypothesized intervening mechanisms. Linear growth models showed significantly greater reduction in boys' delinquency and deviant peer affiliation in the experimental group relative to the controls. Subsequent models using no method overlap in constructs demonstrated that the intervention effect on delinquency operated through growth in parenting and reduction in deviant peer affiliation. PMID- 15819756 TI - Can chimpanzee infants (Pan troglodytes) form categorical representations in the same manner as human infants (Homo sapiens)? AB - We directly compared chimpanzee infants and human infants for categorical representations of three global-like categories (mammals, furniture and vehicles), using the familiarization-novelty preference technique. Neither species received any training during the experiments. We used the time that participants spent looking at the stimulus object while touching it as a measure. During the familiarization phase, participants were presented with four familiarization objects from one of three categories (e.g. mammals). Then, they were tested with a pair of novel objects, one was a familiar-category object and another was a novel-category object (e.g. vehicle) in the test phase. The chimpanzee infants did not show significant habituation, whereas human infants did. However, most important, both species showed significant novelty-preference in the test phase. This indicates that not only human infants, but also chimpanzee infants formed categorical representations of a global-like level. Implications for the shared origins and species-specificity of categorization abilities, and the cognitive operations underlying categorization, are discussed. PMID- 15819757 TI - Localizing cortical sources of event-related potentials in infants' covert orienting. AB - This study used cortical source analysis to locate potential cortical sources of event-related potentials (ERPs) during covert orienting in infants aged 14 and 20 weeks. The infants were tested in a spatial cueing procedure. The reaction time to localize the target showed response facilitation for valid trials relative to invalid or neutral trials. High-density EEG (126 channels) was recorded during the task, and independent component analysis and equivalent current dipole analysis was used to estimate the cortical sources of the EEG during the task. There was a larger P1 ERP component on the valid trials than the other trials (P1 validity effect). The cortical source analysis suggested that this occurred due to activity in Brodmann's areas 18 and 19. A presaccadic ERP component occurred over the frontal scalp areas (-65 ms) and was larger to a target in a cued location than in uncued locations. A potential cortical source for this ERP component was the superior frontal gyrus on the inferior portion of the prefrontal cortex. Increases from 14 to 20 weeks in amplitude of the P1 validity effect and the presaccadic ERP could be modeled by an increase in activation in the corresponding cortical areas. PMID- 15819758 TI - Forming a stable memory representation in the first year of life: why imitation is more than child's play. AB - Although 9-month-old infants are capable of retaining temporally ordered information over long delays, this ability is relatively fragile. It may be possible to facilitate long-term retention by allowing infants to imitate event sequences immediately after their presentation. The effects of imitation on immediate and delayed recognition and on long-term recall were investigated using event-related potentials (ERPs) and elicited imitation, respectively. Mnemonic facilitation resulting from the opportunity to imitate was apparent using both assessments. ERP assessments at immediate and delayed recognition tests suggested that infants who were allowed to imitate had stronger memory representations of familiar stimuli relative to infants who only viewed the presentation of the events. In addition, infants who were allowed to imitate evidenced higher levels of ordered recall after 1 month relative to infants who only watched the experimenter's demonstration. Therefore, imitation proved to have beneficial effects on explicit memory in 9(1/2)-month-olds, providing evidence of its effectiveness as a tool to augment mnemonic capabilities in infancy. PMID- 15819759 TI - Anaesthetists and sedation in the radiology department: involved or left behind? PMID- 15819760 TI - Evaluation of a non-invasive method of assessing opioid induced respiratory depression. AB - Opioid induced respiratory depression is potentially fatal. The aim of this study was to validate a monitoring system that could be used to assess respiratory depression in postoperative patients. The hypercapnic ventilatory response was estimated non-invasively in 12 volunteers. In two steps, we tested a system which delivered carbon dioxide (CO(2)) challenges through a venturi mask, measuring changes in ventilation with an uncalibrated respiratory inductance plethysmograph (RIP). RIP and pneumotachograph measurements of ventilation, taken at the same time during a CO(2) challenge, were similar; group mean (SD), pneumotachograph 13.9 (3.5) l x min(-1) x kPa(-1), RIP 14.3 (2.9) l x min(-1) x kPa(-1). Bland Altmann analysis showed the variation between these two methods was +/- 5 l x min(-1) x kPa(-1) (2 SD). Second, we confirmed that the venturi mask is suitable for delivering CO(2) challenges. Despite the variability in RIP measurements, a simple multimodal respiratory monitoring system could be developed that incorporates clinical observation and non-invasive measurement of the ventilatory response to CO(2). PMID- 15819761 TI - Physostigmine for the prevention of postanaesthetic shivering following general anaesthesia - a placebo-controlled comparison with nefopam. AB - Physostigmine was studied for its efficacy in the prevention of postanaesthetic shivering compared to nefopam and placebo. We studied 89 patients undergoing abdominal and urological surgery. The study was randomised and double-blind, the patients received physostigmine 2 mg (n = 31), nefopam 10 mg (n = 30) or saline (n = 28). Haemodynamic parameters and temperature were measured at induction of anaesthesia (T0), and 5 min (T1), 15 min (T2), 30 min (T3) and 60 min (T4) after reaching the postanaesthetic care unit (PACU). Significantly less shivering occurred following administration of physostigmine and nefopam (9.7 and 3.3%) compared to placebo (53.6%). The degree of shivering was also significantly reduced following physostigmine and nefopam (p < 0.01). Extubation time, haemodynamic parameters and tympanic temperature were found to be similar in all groups. Aldrete score, duration of PACU stay and postoperative analgesic requirements did not differ between the groups. Only nefopam significantly (p < 0.01) reduced postoperative nausea and vomiting. Physostigmine is a safe alternative to nefopam, significantly reducing the incidence and severity of postanaesthetic shivering without affecting postanaesthetic recovery. PMID- 15819762 TI - The effect of calcium channel blockers on outcome following the surgical treatment of phaeochromocytomas and paragangliomas. AB - The peri-operative management of patients undergoing surgery for phaeochromocytoma or paraganglioma with calcium channel blockers (CCB) and their impact on postoperative morbidity and mortality were studied. The medical records of 105 patients undergoing surgery between 1991 and 2002 were analysed retrospectively. In all patients, the calcium channel blocker nicardipine was used for the peri-operative management of haemodynamic changes. Sixty-five patients (61.9%) showed transient intra-operative hypertension. Systolic blood pressure (SBP) > 220 mmHg and SBP > 180 mmHg for > 10 consecutive minutes was observed in 14 (13%) and four patients (2.8%), respectively. SBP < 80 mmHg for > 10 consecutive minutes occurred in 13 patients (12.3%). Eleven patients (10.4%) developed postoperative complications and three patients died (2.8%). The median (range) ICU and hospital length of stay were, respectively, 1 (0-7) day and 10 (2 35) days. The sole use of calcium channel blockers for the peri-operative management of phaeochromocytoma and paraganglioma resection does not prevent all haemodynamic changes; however, its use was associated with a low morbidity and mortality. PMID- 15819763 TI - A comparison of direct laryngoscopy and jaw thrust to aid fibreoptic intubation. AB - We compared two manoeuvres, jaw thrust and laryngoscopy, to open the airway during fibreoptic intubation in 50 patients after induction of anaesthesia in a crossover study. Patients were randomly allocated to receive either jaw thrust or conventional Macintosh laryngoscopy first. Airway clearance was assessed at both the soft palate and the epiglottis. Direct laryngoscopy provided significantly better airway clearance at the level of the soft palate than jaw thrust (44 (88%) vs 31 (62%), respectively; p = 0.002). At the level of the larynx, airway clearance was equally good in both groups (45 (90%) vs 46 (92%), respectively; p = 0.56). The times to view the larynx (median (interquartile range [range]) 4 (3 5 [2-35]) s vs 3 (3-4 [2-8]) s, respectively) and intubation time (20 (17-23 [11 83]) s vs 18 (15-20 [11-28]) s, respectively) were also similar. PMID- 15819764 TI - A comparison of 0.2 and 0.5 mg intrathecal morphine for postoperative analgesia after total knee replacement. AB - The optimal dose of intrathecal morphine for postoperative analgesia after major surgery is a matter of debate, with some uncertainty concerning the therapeutic potential and safety of intrathecal morphine in the dose range 0.3-1.0 mg. This randomised double-blind study compared the efficacy and side-effect profile of 0.2 mg and 0.5 mg intrathecal morphine in 70 patients undergoing knee replacement surgery. The primary endpoint was the number of patients requiring rescue analgesia (tramadol) during the first 24 h postoperatively. Secondary endpoints included consumption of tramadol and the incidence of adverse effects. Fewer patients in the 0.5-mg group required rescue analgesia in the first 24 h than in the 0.2-mg group (16 (48%) vs 28 (85%), respectively; p = 0.003). Median (IQR [range]) tramadol consumption was lower in the 0.5-mg group than in the 0.2-mg group (0 (0-100 [0-350]) mg vs 100 (50-100 [0-350]) mg, respectively; p = 0.02). The incidence of adverse effects was similar in both groups. This study has demonstrated that 0.5 mg intrathecal morphine produces better analgesia than 0.2 mg after knee replacement without any increase in side-effects. PMID- 15819765 TI - A comparison of intrathecal fentanyl and diamorphine as adjuncts in spinal anaesthesia for Caesarean section. AB - This randomised controlled trial examines the effects of fentanyl and diamorphine, alone and in combination, as adjuncts to spinal anaesthesia for Caesarean section. Ninety-nine women undergoing elective Caesarean section with spinal anaesthesia using 0.5% hyperbaric bupivacaine were randomly allocated to receive fentanyl 15 microg (F), diamorphine 0.25 mg (D), or fentanyl 15 microg plus diamorphine 0.25 mg (FD), intrathecally. All women received morphine via a patient controlled analgesia system after surgery. There was no significant difference between the groups in time to achieve a block, discomfort, ephedrine use, nausea and vomiting, pruritus and sedation during surgery. Significant differences were observed in morphine consumption 4, 8, 12 and 24 h after surgery between both F and D groups, and F and FD groups, and also at 2 h between F and FD groups. There was a significant difference in pruritus at 4 h between the F and FD group. Our results suggest that diamorphine alone provides optimum benefits during and after surgery, when used in combination with hyperbaric bupivacaine for Caesarean section. PMID- 15819766 TI - The effect of high altitude commercial air travel on oxygen saturation. AB - Air travel has increased steadily over the last decade, and its effect on the health of passengers has been the subject of much debate. There is a paucity of evidence on the effects of air travel on oxygen saturation in general populations. The peripheral oxygen saturation and pulse rate of 84 passengers, aged 1-78 years, were measured by pulse oximetry at round level and altitude during air travel. There was a statistically significant reduction in oxygen saturation in all passengers travelling long haul and short haul flights (p < 0.05). The mean [range] (SD) SpO(2) for all flights at ground level was 97% [93 100] (1.33) and at cruising altitude 93% [85-98] (2.33). Fifty-four per cent of passengers had SpO(2) values of 94% or less at cruising altitude. This is a value which may prompt physicians to administer supplemental oxygen in hospital patients. PMID- 15819767 TI - Effect of low dose inhaled anaesthetic agents on the ventilatory response to carbon dioxide in humans: a quantitative review. AB - This paper reviews published studies (identified by a Medline-assisted search) on the effect of 6 months post-transplant. Sera were tested using enzyme-linked immunosorbent assay (ELISA) screening for anti-HLA antibodies. Positive sera were retested with ELISA-specific panel for antibody specificity. A 11.2% of patients had anti-HLA antibodies and 4.4% had DSA. Anti-HLA antibodies were significantly associated with pretransplant sensitization, acute rejection and in multivariate analysis, higher serum creatinine (2.15 +/- 0.98 vs. 1.57 +/- 0.69 mg/dl in negative anti-HLA antibodies group). Allograft biopsies performed in a subset of patients with anti-HLA antibodies revealed that 66% had C4d in peritubular capillaries (0% in patients without antibodies). Anti-HLA antibodies were associated with a worse allograft function and in situ evidence of anti-donor humoral alloreactivity. Long-term RTR with an increase in creatinine could be screened for anti-HLA antibodies and C4d in biopsy. PMID- 15819802 TI - Sequential body composition analysis by bioimpedance early post-kidney transplantation. AB - BACKGROUND: While chronic renal failure patients present disturbed body water composition, few studies have been done on its behavior following kidney grafting (Tx). We report the changes associated with a successful Tx on body composition evaluated by bioelectrical impedance analysis (BIA). METHODS: Twelve Tx (seven males, five females) were studied. The BIA was assessed before Tx, at month 1 and at month 3 post-Tx. Total body water (TBW), extracellular water (ECW), intracellular water (ICW), Na:K exchange rate (Nae:Ke) and phase angle (PA) were studied. An healthy group and a HD group were evaluated three times in a year interval. RESULTS: Comparing before Tx with month 1 post-Tx, TBW, ECW and Nae:Ke increased, while ICW and PA decreased significantly. Comparing month 1 with month 3 post-Tx, ECW decreased, while ICW and PA increased. On comparing month 1 post Tx with the healthy group, Nae:Ke was greater and PA was lower at month 1. CONCLUSIONS: The BIA showed that the different body water compartments of Tx recipients quickly match the constitution of normal individuals, overcoming drug therapy side effects. PMID- 15819803 TI - Differences of perioperative liver function, transfusion, and complications according to the type of hepatectomy in living donors. AB - Numerous living donor hepatectomy are being performed safely. However, donors are still exposed to various complications including hepatic failure. We examined the donor's potential risk and morbidity depending on the type of hepatectomy: left lateral segmentectomy (group LLS, n = 30), left lobectomy (group LL, n = 15), and right lobectomy (group RL, n = 128). The charts and computerized hospital data of 173 donors from March 2000 to September 2003 were retrospectively reviewed. We analyzed liver function tests (LFT), RBC transfusion, and complications. Although the graft weight was greatest, and surgical and anesthetic times were longest in the group RL, there were no significant differences in postoperative hospital stay, RBC transfusion, and major complications among the groups. However, minor complications were significantly higher in group RL than group LLS. Postoperative prothrombin time and total bilirubin were significantly higher in the group RL than the other groups (P < 0.05). Living donor hepatectomy is relatively safe, and it is evidenced by rapid recovery of LFT and low occurrence of major complications. However, noticeable depression of LFT and frequent minor complications occur after hepatectomy, especially RL. PMID- 15819804 TI - Angioplasty treatment of hepatic vein stenosis in pediatric liver transplants: long-term results. AB - We reviewed long-term results of percutaneous venoplasty in children with hepatic vein stenosis after partial liver transplants, of which excellent early results were shown. Percutaneous transjugular hepatic venoplasty using balloon dilatation or stent implantation was performed in six cases with hepatic vein stenosis identified on routine post-transplant Doppler sonography and confirmed by transjugular hepatic venography from 1994 to 2003. Repeated procedure was carried out if necessary. Six of 105 patients with partial liver graft developed hepatic stenosis characterized by low hepatic venous velocity with monophasic waveform with significant pressure gradient (>5 mmHg). The incidence was 4.46% for all 112 pediatric liver transplants. Successful balloon venoplasty was achieved in four cases. Self-expanding stent was used in two cases with absent waisting or angulated balloon catheter during dilatation and persisted pressure gradient (>5 mmHg). Repeated procedure was required in two initially successful cases with additional stent used in one case. Three cases had transient hyperdynamic hepatic venous flow with markedly increased central venous pressure after stent implantation. Nonprocedural-related mortality rate was 16.7%. Patent hepatic vein was maintained in five patients after a mean follow-up of 3.67 years (0.75-9.5). Higher incidence of hepatic vein stenosis was noted in pediatric partial liver transplant. However, encouraging long-term results showed that hepatic venoplasty or stent implantation could be a preferable alterative to surgical revision or retransplantation, which has been the procedure of choice in our hospital. PMID- 15819805 TI - Bronchoscopy in the diagnosis and surveillance of respiratory infections in lung and heart-lung transplant recipients. AB - Fiberoptic bronchoscopy (FOB) with bronchoalveolar lavage (BAL) and transbronchial biopsies (TBB) is a widely used method to detect respiratory infections and to differentiate them from other postoperative complications in lung transplant (LTX) recipients, but the usefulness of surveillance FOBs is not yet established. The aim of this study was to evaluate the usefulness of FOB in the diagnosis and surveillance of infections in LTX recipients. We reviewed all the consecutive 609 FOBs performed on 40 lung or heart-LTX recipients between February 1994 and November 2002. The overall diagnostic yield was 115/190 (61%) and 43/282 (15%) for clinically indicated and surveillance FOBs respectively (P < 0.001). Infection was established by bronchoscopic samples in 96/190 (50.5.%) of the clinically indicated FOBs and 34/282 (12.1%) of the surveillance FOBs (P < 0.001). The diagnostic yield of the clinically indicated FOBs was highest (72%) from 1 to 6 months post-transplant (P = 0.04). Pneumocystis carinii was detected in 23 (4.9%) of the bronchoscopic specimens and 15 (65%) of the P. carinii infections were detected during adequate chemoprophylaxis. To conclude, in LTX recipients clinically indicated FOB has a good diagnostic yield in detecting infections and other postoperative complications, whereas the information received from surveillance FOB has remained less significant. With current prophylaxis and screening strategies FOB is still required to detect P. carinii infections. PMID- 15819806 TI - Predictors of death on the waiting list for liver transplantation characterized by a long waiting time. AB - The number of patients dying while on the liver transplantation (LT) waiting list (WL) has continued to increase in recent years as a result of severe shortage of organs. Therefore, it is important to evaluate the existing models that predict death on the WL and to determine the independent predictors of death. The study cohort comprised 152 adult patients listed for LT in our centre over a period of 2 years (January 2001 to January 2003). The 12-month survival rate has been calculated by Kaplan-Meier method. The survival analysis performed by Cox proportional hazard model has evaluated the three parameters which compose the model for end-stage liver disease (MELD) score. Forty-four patients (28.9%) died while listed for LT. The survival rate was 92% at 3 months, 80% at 6 months and 69% at 12 months. Median survival was not reached. MELD score was found to be an excellent predictor of death at 12 months on our WL--c-statistic (area under curve) 0.84. In our survival analysis, only international normalized (prothrombin) ratio (INR) and serum creatinine were identified as an independent predictors of death (P < 0.0001). A new simplified version of the MELD score, which does not include serum bilirubin, is proposed and its c-statistic as predictor for death on the WL at 12 months is 0.86, as good as the original MELD score, when evaluated on our list. There is a fourfold increase in mortality on our WL for LT between 3 and 12 months after the inclusion. A simplified version of the MELD score, using only serum creatinine and INR might be taken into account when predicting 12 months mortality on WL with longer waiting time, but it has to be confirmed by other prospective studies. PMID- 15819807 TI - Assignment of steatotic livers by the Mayo model for end-stage liver disease. AB - Prognosis after liver transplantation depends on a combination of recipient and donor variables. The purpose of this study is to define an allocation system of steatotic donor livers relative to recipient model for end-stage liver disease (MELD) score. We reviewed 500 consecutive OLT, computing the MELD score for each recipient. Fatty infiltration in grafts was categorized in no steatosis, 10-30%, 30-60% and > or = 60% steatosis. MELD score did not affect preservation injury and graft dysfunction, which were increased with fat content. Recipient and graft survivals lowered when increasing MELD score. Outcome in low-risk recipients (MELD < or = 9) was not altered with steatosis, except those with > or = 60%. Survival functions in moderate-risk recipients (MELD 10-19) were moderately affected with 10-30% steatosis and severely with those with >30. Exactly 30-60% steatotic grafts work poorly in high-risk recipients (MELD > or = 20), and very poorly with > or = 60% steatosis. Prognosis of candidates is optimally influenced when divergence of recipient-donor risks is presented. PMID- 15819808 TI - Microbial surveillance during human pancreatic islet isolation. AB - The aim of the study was to investigate microbiological contamination rate during human pancreatic islet isolation. Between 1996 and 2002, pancreas preservation media and post-purification islet preparations were screened for microbiological contamination. After arrival in the laboratory, pancreata were washed prior to enzyme perfusion with either Hank's balanced salt solution (Group I, n = 170, 1996 to 2001) or decontaminated with polyvidonum-iodine, cefazoline, and amphotericine B (Group II, n = 45, 2001 to 2002). Microbiological contamination of preservation media was observed in 56% and 84% for Groups I and II, respectively. Analysis of contaminants revealed 74% Gram-positive, 21% Gram negative bacteria and 5% fungi. Duration of transport had an influence on the rate of contamination (P < 0.05). After islet isolation, Group I presented microbial contamination of 16 islet preparations (9.4%) [i.e. Gram-positive bacteria (n = 10), Gram-negative bacteria (n = 4), and fungi (n = 2)]. In Group II, only 2 islet preparations (4.4%) presented microbial contamination. Microbial contamination during pancreas procurement occurs frequently. Most microorganisms are eliminated during islet isolation, and de novo contaminations during islet isolation are rare. Pancreas decontamination reduces the risk of infection of the final islet preparation. PMID- 15819809 TI - Immunosuppressive efficacy of mycophenolate mofetil when compared with azathioprine and mizoribine against peripheral lymphocytes from renal transplant recipients. AB - Mycophenolate mofetil is currently used instead of azathioprine in clinical transplantation. However, comparative studies for the immunosuppressive potency of anti-metabolites used for organ transplantation have not been well documented. We compared the pharmacological efficacy of mycophenolic acid (MPA), 6 meraputopurine (6-MP), and mizoribine (MZ) for inhibiting purine synthesis of peripheral blood mononuclear cells (PBMCs) in vitro by a mitogen assay procedure. PBMCs were obtained from 18 renal transplant recipients before operation and 18 healthy subjects. The inhibitory efficacy of 6-MP against concanavalin A-induced PBMC blastogenesis exhibited large variations between subjects in both recipients and healthy subjects. In contrast, the pharmacological efficacy of MPA on PBMC blastogenesis showed the smallest inter-individual variation of all the purine synthesis inhibitors examined. Furthermore, the effects of MPA were almost similar in the recipients and healthy subjects. The pharmacological efficacy of MZ against PBMC blastogenesis was weaker than that of the other two agents and the inter-individual variation of MZ IC50 against PBMCs of the patients was larger than that of MZ IC50 against PBMCs of healthy subjects. Reproducible immunosuppressive efficacy of MPA compared with other purinesynthesis inhibitors could be expected from the viewpoint of MPA pharmacodynamics against PBMCs in renal transplantation. PMID- 15819811 TI - Role of protein synthesis in the protection conferred by ozone-oxidative preconditioning in hepatic ischaemia/reperfusion. AB - The liver is damaged by sustained ischaemia during liver transplantation, and the reperfusion after ischaemia results in further functional impairment. Ozone oxidative preconditioning (OzoneOP) protected the liver against ischaemia/reperfusion (I/R) injury through different mechanisms. The aim of this study was to investigate the influence of the inhibition of protein synthesis on the protective actions conferred by OzoneOP in hepatic I/R. Rats were treated with cycloheximide (CHX) in order to promote protein synthesis inhibition after OzoneOP treatment. Plasma transaminases, malondialdehyde and 4-hydroxyalkenals and morphological characteristics were measured as an index of hepatocellular damage; Cu/Zn-superoxide dismutase (SOD), Mn-SOD, catalase, total hydroperoxides and glutathione levels as markers of endogenous antioxidant system. OzoneOP increased Mn-SOD isoform and ameliorated mitochondrial damage. CHX abrogated the protection conferred by OzonoOP and decreased Mn-SOD activity. Cellular redox balance disappeared when CHX was introduced. Protein synthesis is involved in the protective mechanisms mediated by OzoneOP. Ozone treatment preserved mitochondrial functions and cellular redox balance. PMID- 15819810 TI - Use of different immunosuppressive strategies in recipients of kidneys from nonheart-beating donors. AB - In nonheart-beating donor (NHBD) kidney transplants, immunosuppressive management is difficult mainly because of the high incidence of acute tubular necrosis. This has meant that since the start of our NHBD transplant program, several immunosuppression regimes have been used. The aim of this retrospective study was to evaluate the results obtained over 7 years using different treatment protocols. A total of 172 consecutive NHBD transplants performed between April 1996 and December 2002 were treated as follows: G-I (n = 21), cyclosporine (8 mg/kg/day) plus azathioprine plus steroids; G-II (n = 65), low-dose cyclosporine (5 mg/kg/day) plus mycophenolate plus steroids; G-III (n =17), low-dose tacrolimus (0.1 mg/kg/day) plus mycophenolate plus steroids; and G-IV (n = 69), daclizumab plus low-dose tacrolimus plus mycophenolate plus steroids. Delayed graft function rates were 76.2%, 72.3%, 76.5%, and 42%, respectively, for the four groups (P = 0.000). Rejection-free patient rates were 76.2%, 46.2%, 35.3%, and 71% (P < 0.001). Vascular rejection rates were 19%, 30.8%, 52.9%, and 18.8%, (P = 0.025). Two-year graft survival was 71.4% in group I, 95.4% in group II, 94.1 in group III, and 93.8% in group IV (P =0.004). Patient survival was worse in group I (75.2% in group I, 100% in group II, 100% in group III, and 96.7% in group IV at 2 years; P < 0.001). The use of daclizumab and low-dose tacrolimus could be effective at lowering the incidence of delayed graft function in NHBDT, with no negative repercussions on acute rejection. PMID- 15819812 TI - Fusarium peritonitis concomitant to kidney transplantation successfully managed with voriconazole: case report and review of the literature. AB - Fusarium infections in solid organ transplant recipients are often localized, occur later in the post-transplantation period, and have a better outcome than fusarial infections in patients with hematologic malignancies or bone marrow transplants. We report the first case of proven peritonitis caused by Fusarium species in a renal transplant recipient which is also the first successfully managed with voriconazole. We also review previously reported cases of fusarial infection in solid organ transplant recipients. PMID- 15819813 TI - Effects of pancreas-kidney transplantation on diabetic retinopathy. AB - The effects of pancreas transplantation (PTx) on diabetic retinopathy (DR) are still debated. We studied the course of DR in 48 patients (age: 40 +/- 7 years; males/females 26/22, body mass index (BMI): 23.0 +/- 2.4 kg/m2, duration of diabetes: 24 +/- 8 years) bearing a successful PTx (combined with a kidney). Follow-up ranged 6-60 months (median: 17 months). Before transplantation, according to the Eurodiab Study classification, 12 patients (25%) had nonproliferative retinopathy (NPDR; mild, moderate or severe), and 36 patients (75%) had laser-treated and/or proliferative retinopathy (LT/PDR). During the follow-up, in the NPDR group improvement/deterioration was defined as regression/progression to a lower/higher retinopathy grade; in the LT/PTD group, stabilization was defined as no new neo-vessel formation or development of new lesions requiring laser-treatment. In the NPDR group, five (41.7%) patients improved of one or more lesion grading, three (25%) patients showed no change, and four (33.3%) patients progressed of one grade. In the LT/PDR group, the post transplant data were: stabilization in 35 (97%) patients, and worsening in one (3%) patient. The number of improved/stabilized patients was significantly higher in the transplanted than in a control group of nontransplanted type 1 diabetic patients. In conclusion, despite a relatively short follow-up period, successful PTx in our cohort of patients was associated with improvement and/or stabilization of DR in the majority of recipients. PMID- 15819815 TI - Sjogren's syndrome. AB - Sjogren's syndrome (SS) is a chronic autoimmune disease affecting the exocrine glands, primarily the salivary and lacrimal glands. It has been suggested that exogenous agents may trigger SS in genetically predisposed individuals. However, at present, the etiology of SS is far from being understood, and no direct evidence for any of these triggers has been presented. The salivary and lacrimal glands from patients with SS harbor unique and highly selected T- and B-cell populations. Disturbance in glandular cell apoptosis may be one possible explanation for the sicca symptoms in SS. However, discrepancies between glandular destruction and salivary flow give rise to processes causing glandular dysfunction preceding or triggering glandular cell destruction. Recent reports suggested autoantibodies inhibiting neuronal innervation of acinar cells and defective water transport to be implicated in salivary secretion deficiency observed in SS. Several types of autoantibodies have been suggested to contribute to the pathogenesis of SS. However, how the tolerance to these structures is broken down is unknown at present. Studies on B-cell activating factor indicated that diminished apoptosis and disturbed B-cell maturation could be responsible for the occurrence of autoreactive B-cells and B-cell hyperreactivity. B-cell activation may also provide a basis for lymphoma development observed in up to 5% of the patients with SS. PMID- 15819814 TI - Severe azathioprine-induced myelotoxicity in a kidney transplant patient with thiopurine S-methyltransferase-deficient genotype (TPMT*3A/*3C). PMID- 15819816 TI - Changes in the bucco-lingual thickness of the mandibular alveolar process and skeletal bone mineral density in dentate women: a 5-yr prospective study. AB - After tooth extraction there is a great interindividual variation in the remodelling pattern of the alveolar process in edentulous areas, with some individuals losing little bone and others undergoing extensive resorption. However, little is known about possible longitudinal changes in the dentate region of the alveolar process of adults and if these are related to alterations in the skeletal bone mineral density (BMD). In a prospective study, on two occasions, 5-yr apart, the BMD of 117 women was determined in the distal forearm by using dual-energy X-ray absorptiometry, and the bucco-lingual thickness of the mandibular alveolar process was measured on dental casts by using a dial calliper. A decrease in the mean alveolar thickness, exceeding a cut-off value of 0.1 mm, was found in 60% of the women and an increase was found in 3% of the individuals. This decrease was 0.22 +/- 0.20 mm in the posterior region and 0.16 +/- 0.19 mm in the anterior region. The changes in alveolar thickness in the posterior region were significantly correlated to the BMD changes both on the mid crestal level site and on the cervical level site. We conclude that the bucco lingual thickness decreases with age in the dentate alveolar process, possibly owing to periosteal resorption related to skeletal bone loss. PMID- 15819817 TI - Sense of coherence as a determinant of the oral health-related quality of life: a national study in Finnish adults. AB - Our aim was to investigate the relationship among the oral health-related quality of life, the sense of coherence (SOC), and socio-economic and demographic factors, oral behavioural variables and oral health variables. The sample of this study consisted of 4,039, 30-64-yr-old dentate adults (1,899 men and 2,140 women). The questionnaire and home interview included information about socio economic and demographic factors, behavioural and attitudinal variables (such as oral health behaviours and oral health variables), the SOC (12-item) and the Oral Health Impact Profile scales (OHIP-14). Subjects with a strong or moderate SOC were found to have significantly fewer problems attributed to oral conditions (OHIP) than those with a weak SOC. The SOC was also associated with all of the subscales of the OHIP, and the association was most evident in psychological discomfort, psychological disability and handicap subscales. The SOC appears to be a determinant of the OHIP, independently of oral health, oral health behaviour and socio-economic factors. A psycho-social aspect is strongly embodied in the oral health-related quality of life of individuals. PMID- 15819818 TI - Vascular status in human primary and permanent teeth in health and disease. AB - The present study sought to compare the vascular status of human primary teeth with that of human permanent teeth, and to determine whether caries or painful pulpitis was associated with changes in vascularity. Coronal pulps were removed from 62 primary and 62 permanent mandibular molars with a known pain history. Teeth were categorized as intact, moderately carious or grossly carious. Pulp sections were labelled with Ulex europaeus I lectin (UEIL), which is a marker of human vascular endothelium. Image analysis was then used to quantify the percentage area of UEIL-labelled tissue (vascularity) and the number of blood vessels present within three regions: the pulp horn, the subodontoblastic region, and the mid-coronal pulp. Only the mid-coronal region of the primary tooth pulp was found to be significantly more vascular than the corresponding area of the permanent tooth pulp. Both dentitions showed a significant increase in vascularity within the pulp horn region with caries progression, but this was not accompanied by an increase in vessel number. There was no correlation between vascularity and pain symptoms. These findings suggest that the primary tooth pulp is more vascular than its successor within the mid-coronal region. However, the functional and clinical significance of this finding remains speculative. PMID- 15819819 TI - The accuracy of individual self-reported items to determine periodontal disease history. AB - The objective of this study was to evaluate the validity of self-reported periodontal disease and symptoms to predict periodontal disease history in a German population. We analysed data from 246 patients who answered a self administered questionnaire on self-perceived periodontal disease and self reported symptoms of periodontal disease. Actual periodontal status was assessed from panoramic radiographs and periodontal disease was defined as > or = 3 teeth with radiographic alveolar bone loss (ABL) > 5 mm (prevalence 39%). Sensitivity (SN) and specificity (SP) of self-reported items for diagnosis of periodontal disease history were calculated. The diagnostic accuracy of any individual item was generally low. For example, self-perceived periodontal disease had SN of 49% and SP of 67%. All self-reported items had low sensitivity, while self-reported professionally diagnosed bone loss, tooth loss caused by periodontal disease and mobility had SP > 90%. In conclusion, we did not identify a single individual question that can assess periodontal disease from self-reporting with satisfactory validity. Future studies will evaluate whether multivariate prediction rules using information from several self-reported variables are a valid means to ascertain periodontal disease history. PMID- 15819820 TI - The influence of toothpastes and prophylaxis pastes on fluorescence measurements for caries detection in vitro. AB - The aim of this study was to test the influence of toothpaste and prophylaxis paste remnants as well as powder remnants on DIAGNOdent readings. A total of 68 sites on the occlusal surface of human molars were used for this study. After a cleaning procedure, saliva was spread on the tooth surface, dried for 3 s, and DIAGNOdent (DD) values were measured. After these measurements were performed, 500 mg of the toothpaste or prophylaxis paste was taken and the occlusal surface was cleaned for 3 s with a slow rotating contra-angle handpiece. The paste was then rinsed off with the 3-in-1 syringe for 3 s using water with air. The DD measurements were taken again and the rinsing for 3 s and fluorescence measurements were repeated. The procedure with the powder using the PROPHYflex 2 was, in principle, the same. Whereas only small differences could be detected on sound surfaces, there was a distinct influence on decayed surfaces. It could be concluded that when the DIAGNOdent device is used in practice and special care in rinsing after cleaning is not taken, it is possible that false readings are registered. PMID- 15819821 TI - Multivariate survival analysis for the identification of factors associated with cavity formation in permanent first molars. AB - This investigation used a multivariate survival model, which allowed for dependent caries data with possible censoring, to analyse the impact of timing of tooth emergence, gender and several reported oral hygiene and dietary habits on the incidence of cavity formation in permanent first molars (PFM) in young children. A 6-yr prospective oral health screening project in Flanders, Belgium, provided clinical and questionnaire data on 4,468 children. The results revealed that PFM in children who reported frequent brushing, who had no visible caries experience in the primary dentition, and who presented without occlusal plaque on the PFM had the best survival estimates. Girls had a higher incidence than boys of cavity formation in mandibular molars. The multiple survival analysis confirmed the major impact of the caries status of the deciduous dentition and self-reported oral hygiene habits on the incidence of cavities in permanent first molars. PMID- 15819822 TI - Dynamic protein expression patterns during intraoral wound healing in the rat. AB - Wound healing after cleft palate surgery is often associated with impairment of maxillary growth and dento-alveolar development. Wound contraction and scar tissue formation contribute strongly to these effects. In vitro studies have revealed that fibroblasts isolated during different phases of palatal wound healing show phenotypical differences. They change from a quiescent to an activated state and then partly back to a quiescent state. In this study, we evaluated the existence of fibroblast phenotypes at several time-points during palatal wound healing in the rat. Based on cytoskeletal changes (alpha-sma, vimentin, vinculin), integrin expression (alpha1, alpha2, alpha(v) and beta1) and changes in cellularity, we conclude that phenotypically different fibroblast populations are also present during in vivo wound healing. Alpha-sma and the integrin subunits alpha1 and alpha(v) were significantly up-regulated, and vinculin was significantly down-regulated, at early time-points compared to late time-points in wound healing. These changes point to an activated fibroblast state early in wound healing. Later in wound healing, these activated fibroblasts return only partially to the unwounded situation. These results strongly support the idea that different fibroblast populations with specific phenotypes occur in the course of palatal wound healing. PMID- 15819823 TI - Assessment of dental appearance following changes in incisor proportions. AB - This study aimed to evaluate the assessment of attractiveness of standardized changes in incisor proportions. Two original photographs, showing the maxillary anterior teeth in the frame of a smile, were computer-manipulated. From the so produced symmetrical 'golden standard' images, two sets were constructed. Each set contained one golden standard image and seven images with different standardized changes concerning (i) width-to-length ratios of the central incisors (60-96%) and (ii) tooth-to-tooth proportions between the widths of lateral and central incisors (43-87%). The judges, consisting of three groups of individuals (24 dentists, 24 medical students, and 179 patients), ranked each photo set for attractiveness on a visual analogue scale. The width-to-length ratios were assessed as most attractive within a range of 75-85% (medical students, patients) or 75-80% (dentists), respectively. The tooth-to-tooth proportions showed the best results concerning attractive appearance within a range of 50-74% (medical students, patients) or 56-68% (dentists), respectively. No significant differences between the participant groups were found except for some extreme variations. It was concluded that a most attractive 'golden range' could be identified for both the width-to-length ratios and the tooth-to-tooth proportions of the maxillary incisors. PMID- 15819824 TI - Plasticizers in denture soft-lining materials: leaching and biodegradation. AB - The leaching from soft lining materials into an aqueous buffer with the same esterase activity as that of saliva was compared to leaching into buffer without esterase activity. The buffer contained 0.1% non-ionic detergent, giving the medium a capacity to dissolve phthalates to the same degree as saliva. The hypothesis that esterase in the immersion medium will increase the rate of diffusion of plasticizers from denture soft-lining material was confirmed by the results. The average leaching of phthalates from 5.5 g of one of the materials was 4.5 mg kg(-1) d(-1) within the first 2 d and 1.1 mg kg(-1) d(-1) within the first 28 d. These levels may be compared to a LOAEL (lowest observed adverse effect level) of 52 mg kg(-1). It is concluded that an esterase activity, equivalent to that in saliva, in the immersion medium for soft lining materials increased the rate of diffusion of plasticizer from the materials. The measured levels of phthalates leaching from these materials might in vivo only be slightly less than 1/10 of the LOAEL given above. PMID- 15819825 TI - Colour and translucency of opaque-shades and body-shades of resin composites. AB - The purpose of this study was to evaluate the optical properties--not only the translucency but also the colours--of opaque-shade resin composites. The CIELAB parameters (L*, a* and b*) of disks of A2 and opaque A2 (OA2) shades of Charisma (Heraeus-Kulzer), Solare (GC) and Filtek Supreme (3M) were evaluated on backings of black, white and the material itself to calculate the translucency parameter (TP) and the colour differences (delta E*) between A2 and OA2. A two-way analysis of variance (anova) for the TP indicated a less statistically significant TP value in the OA2 shade than the A2 shade for all products. As for the products, Charisma showed a statistically greater TP value than the other two products. Regarding the delta E* between A2 and OA2, all the products revealed clinically perceptible colour differences (delta E* > 3.3). Hence, we must take the colour differences of opaque-shade resin composites into consideration, as well as the translucency of the materials, for a clinically acceptable colour match of the restoration. PMID- 15819826 TI - Clinical fit of all-ceramic three-unit fixed partial dentures, generated with three different CAD/CAM systems. AB - In this study, the hypothesis was tested that the marginal and internal fit of CAD/CAM fabricated all-ceramic three-unit fixed partial dentures (FPDs) can be as good as in metal-ceramic FPDs. Twenty-four all-ceramic FPDs were fabricated and randomly subdivided into three equally sized groups. Eight frameworks were fabricated using the Digident CAD/CAM system (DIGI), another eight frameworks using the Cerec Inlab system (INLA). Vita Inceram Zirkonia blanks were used for both groups. In a third group frameworks were milled from yttrium-stabilized Zirconium blanks using the Lava system (LAVA). All frameworks were layered with ceramic veneering material. In addition, six three-unit metal-ceramic FPDs served as control group. All FPDs were evaluated using a replica technique with a light body silicone stabilized with a heavy body material. The replica samples were examined under microscope. The medians of marginal gaps were 75 microm for DIGI, 65 microm for LAVA and INLA and 54 microm for the conventional FPDs. Only the DIGI data differed significantly from those of the conventional FPDs. Within the limits of this study, the results suggest that the accuracy of CAD/CAM generated three-unit FPDs is satisfactory for clinical use. PMID- 15819827 TI - X-ray absorption fine structure analysis of the local environment of zinc in dentine treated with zinc compounds. AB - It has been reported that zinc oxide (ZnO) inhibits dentine demineralization. By using the X-ray absorption fine structure (XAFS) technique, our aims in this study were to provide information about the local environment of zinc atoms in dentine that had been treated with zinc compounds. We measured the Zn K-edge X ray absorption near-edge structure (XANES) and the extended X-ray absorption fine structure (EXAFS) of dentine specimens treated with zinc chloride or ZnO. In XAFS analyses, the spectra of dentine specimens treated with ZnO (D-ZO) or with zinc chloride (D-ZC) were similar and obviously different from the reference ZnO spectrum. This suggests that most of the zinc atoms detected in D-ZO are not derived from particles of ZnO. The spectra of D-ZO and D-ZC were similar to the spectrum of the synthetic, zinc-containing hydroxyapatite, but were not similar to that of zinc in ZnCl2-treated collagen. The results of this study suggest that most of the zinc atoms detected were attached to hydroxyapatite and not to collagen. PMID- 15819829 TI - Comment on "A retrospective study of dental behaviour management problems in children with attention and learning problems". PMID- 15819831 TI - Complex nursing systems. AB - The specific aim of this paper is to identify the causes underlying the growth in health care system complexity, discuss challenges nurse administrators face as a result of it and suggest strategies for better decision-making in complex system environments. This paper further discusses the merits of computational modelling and simulation which can assist nurse administrators predict the results of their decisions through a virtual complex system environment. PMID- 15819832 TI - Recording the content of the caring process. AB - In the present study, a theoretical basis for the content of caring and nursing documentation is described. The goal was to find out to what extent documentation based on the theory might reveal the patients' experience and views of their care. Documenting according to Eriksson's (1997, 2001) caring science theory was tried out in an intervention study. The content of nursing care records was evaluated before the intervention and after completing it. After the intervention the nurses' experiences of the theory-based recording were collected by means of questionnaires. According to the evaluation performed after the intervention, the content of the documentation had improved on several wards. The nurses paid more attention to the patients' views than before when documenting care in the way introduced in the study. According to the results nurses need strong support from their managers in order to successfully be able to implement a theoretical basis of documentation of care. PMID- 15819833 TI - First-line managers' views of the long-term effects of clinical supervision: how does clinical supervision support and develop leadership in health care? AB - There have recently been several organizational changes that have challenged nursing managers in the Finnish health care system. First-line managers need support in their work because of organizational changes and scarce economic resources. One of these supportive measures is clinical supervision. A group of first-line managers in a Finnish University hospital participated in a 2-year clinical supervision intervention in 1999-2000. The managers' perceptions of the clinical supervision were followed up twice during the intervention and 1 year after (2001). The aim of this study is to describe how the first-line managers saw the future effects of the clinical supervision intervention 1 year after its termination. At the beginning of the intervention, the number of participating nursing managers was 32. The number of respondents in this study 1 year (2001) after the clinical supervision was 11. Data was collected using empathy-based stories, which involved writing short essays. The respondents received orientation and a script to assist them in the writing of essays. The stories were analysed qualitatively by categorizing the responses by themes. The managers deemed that clinical supervision had, in the 3-year time frame, positive long term effects on their leadership and communication skills, the desire for self development, self-knowledge and coping. Managers believed that in the long run, clinical supervision would provide them with a broader perspective on work and would enhance the use of clinical supervision as a supportive measure among co workers. First-line managers expect clinical supervision to have long-term positive effects on their work and coping. Empathy-based stories, as a method, were found suited to studies, which aim to obtaining future-oriented knowledge. PMID- 15819834 TI - Nurses' satisfaction with their work environment and the outcomes of clinical nursing supervision on nurses' experiences of well-being -- a Norwegian study. AB - BACKGROUND: Various studies have demonstrated that nursing is stressful and that the incidence of occupational stress-related burnout in the profession is high. AIM: This descriptive-correlational study examined nurses' satisfaction with their psychosocial work environment, their moral sensitivity and differences in outcomes of clinical nursing supervision in relation to nurses' well-being by systematically comparing supervised and unsupervised nurses. METHODS: Nurses were selected from two hospitals (n = 71). Data collection was by means of questionnaires and analysed by descriptive and inferential statistics. RESULTS: The nurses' satisfaction with their psychosocial work environment was reflected in six factors: 'job stress and anxiety', 'relationship with colleagues', 'collaboration and good communication', 'job motivation', 'work demands' and 'professional development'. The nurses' perceptions of moral sensitivity comprised seven factors: 'grounds for actions', 'ethical conflicts', 'values in care', 'independence patient-oriented care', 'the desire to provide high-quality care' and 'the desire to provide high-quality care creates ethical dilemmas'. Nurses well-being were reflected in four factors 'physical symptom and anxiety', 'feelings of not being in control', 'engagement and motivation' and 'eye strain sleep disturbance'. The moral sensitivity 'ethical conflicts' were found to have mild negative correlations with psychosocial work environment 'job stress and anxiety professional development' and with 'total score' psychosocial work, moral sensitivity factor 'independence were correlated with psychosocial work factor 'relationships with colleagues' and 'total score', moral sensitivity were mildly correlated with 'collaboration and good communication and had a negative correlation to psychosocial work factor 'work demands'. In addition, significant correlations were found between the nurses' well-being profile and demographic variables, between 'engagement and motivation' and 'absence due to illness' and between 'time allocation for tasks', 'physical symptoms and anxiety' and 'age'. Mild significant differences were found between nurses attending and not attending group supervision and between 'physical symptoms and anxiety' and 'feelings of not being in control'. CONCLUSIONS: We conclude that ethical conflicts in nursing are a source of job-related stress and anxiety. The outcome of supporting nurses by clinical nursing supervision may have a positive influence on their perceptions of well-being. clinical nursing supervision have a positive effect on nurses physical symptoms and their feeling of anxiety as well as having a sense of being in control of the situation. We also conclude that psychosocial work have an influence on nurses experience of having or not having control and their engagement and motivation. PMID- 15819835 TI - Factors influencing nurse supervisor competence: a critical incident analysis study. AB - AIM: The aim of the study was to identify factors related to critical incidents that influence the competence of nurse supervisors. BACKGROUND: Nurse supervisors require considerable competence in order to help supervisees to reflect on their clinical work and to interpret the needs of the patient. METHOD: A qualitative approach involving the critical incident technique was used. Critical incidents were collected by means of self-reports from 25 nurse supervisors. FINDINGS: Two main areas emerged: a professional and a personal stance. The professional stance described the nurse supervisors' awareness of the importance of creating a secure learning environment and facilitating reflection. The supervisors structured the material and created awareness of fundamental nursing values. The second main area, personal stance, described the nurse supervisors' behaviour when they gave the participating nurses the opportunity to work through the experiences gained in the daily provision of nursing care. Although they experienced lack of self assurance during the supervision session, they also expressed security regarding their own performance as nurse supervisors. CONCLUSIONS: Nurse supervisors need to include more nursing theory and focus on the nursing process as well as being aware of their own shortcomings and resources. One way for the supervisor to scrutinize his/her actions is to discuss and examine them with a more experienced nurse supervisor colleague. PMID- 15819836 TI - The influence of standardized languages on nurses' autonomy. AB - AIM: To investigate the influence of standardized languages on nurses' autonomy. BACKGROUND: The uses of standardized languages are expanding. These languages unify communication across health care settings. Autonomy is a favourable concept for nurses. The use of standardized languages may hinder nurses' autonomy; however, these languages may also promote nurses' autonomy. EVALUATION: Review of the literature. KEY ISSUES: The key issue discussed in this paper is whether the use of standardized languages affects nurses' autonomy. CONCLUSIONS: Nurses should use standardized languages in every day practices as it reflects a mastery over their own work. Having control over practice is an important attribute of nurses' autonomy. PMID- 15819837 TI - Stress and verbal abuse in nursing: do burned out nurses eat their young? AB - AIM: The purpose of this study was to explore the types and frequency of verbal abuse of nurses by other nurses. Further, this study explored the components, characteristics, consequences and effects of abuse in an effort to better understand the dynamics of verbal abuse of nurses in the workplace. Nurses who experience occupational burnout are more likely to abuse other nurses. BACKGROUND: It is believed that nurses have been historically subjected to verbal abuse by colleagues, something previously considered to be done primarily by doctors. The effects and consequences of verbal abuse can be devastating and long lasting. Research that has focused on its consequences has found that it is both physiologically and psychologically damaging. METHOD: Participants completed an adapted survey, incorporating the Verbal Abuse Scale and the Verbal Abuse Survey and demographic questions developed by the researchers. Specifically, types of verbal aggression, the frequency and stressfulness of each type, emotional reaction to verbal aggression, cognitive appraisal of verbally aggressive encounters, and similarity and effectiveness of coping behaviours were explored. The long-term negative effects of verbal aggression, including absenteeism and errors in patient treatment, were also evaluated to determine if verbal aggression is a contributing factor. RESULTS: Respondents reported that the most frequent source of abuse was nurses (27%), followed by patients' families (25%), doctors (22%), patients (17%), residents (4%), other (3%) and interns (2%). Of those who selected a nurse as the most frequent source, staff nurses were reported to be the most frequent nursing source (80%) followed by nurse managers (20%). CONCLUSIONS: Verbal abuse in nursing is quite costly to the individual nurses, the hospitals and the patients. Nurses who regularly experience verbal abuse may be more stressed, may feel less satisfied with their jobs, may miss more work and may provide a substandard quality of care to patients. PMID- 15819838 TI - Refocusing health visiting -- sharpening the vision and facilitating the process. AB - AIM: The purpose of this study was to explore the development of public health nursing in a primary care trust that focused around the evaluation of a newly introduced public health nursing role. The intention was to inform role development during the three year evaluation period. BACKGROUND: Despite health visitors having been identified in the recent past as being in prime position to assume the public health role envisioned by the British Government in the New NHS, the vision for the 'new look' health visitor has been rather blurred. METHODS: An action research approach utilised 22 individual and four group (number of participants 33) interviews with a range of stakeholders. Thematic analysis was used on all transcripts. FINDINGS: Four main headings were identified to organise and present the findings -- definitions of public health; continuum of practice; contextual factors in public health development; and inhibitors to public health practice development. The findings suggest that there are a number of factors challenging the health visiting response to the public health agenda. Facilitating clarity of role and purpose must be a management priority. A tool, developed from this study, offers a means to explore practice within a primary care -- public health continuum and provides a schema against which to set and action practice development. PMID- 15819839 TI - Bridging the information gap between hospitals and home care services: experience with a patient admission and discharge form. AB - Hospital care and ambulatory care are institutionally and financially so deeply separated in Germany that discontinuity of individual treatment and, hence, losses in both the quality and efficiency of care are all too often the consequences of this widely deplored systemic defect. In order to improve the communication between home care services and hospitals during admission or discharge of patients in need of long-term nursing care, the research project 'aski' has developed an innovative instrument and procedure for the reciprocal transfer of information between these institutions. After successful testing, the Patient Accompanying Form was offered for public use. Two years later, a written and telephone survey was conducted among the somewhat disappointingly small number of its users. Based on their experience, issues surrounding the implementation of such instruments in the health care system are discussed as well as the chances of enhancing integrative care by promoting coordinated communication between the providers. PMID- 15819840 TI - Estimating resource use and cost of prophylactic management of neutropenia with filgrastim. AB - The study objective is to develop a methodology for the measurement of time, resource use and cost of the prophylactic management of neutropenia with filgrastim in different settings where the drug is routinely used: in-hospital care, outpatient care and home care. The activity-based costing method is used to analyse the cost of managing prophylactically neutropenia and comprises four steps. First, department heads in each of the chosen settings were selected and interviewed to obtain key elements in the workflow that involves the management of neutropenia, followed by the second step involving in-depth, structured interviews of key personnel. The third step was the measurement of the time required for frequently occurring activities in monitoring neutropenia and the administration of filgrastim by a study nurse. Finally, information on resource unit costs and personnel salaries were collected from the administration units to calculate an average cost. Sensitivity analyses were undertaken on estimated variables in the study. A list of eight to 14 consecutive activities linked to the prophylactic management of neutropenia was observed. The number and type of activities do not differ between an in-hospital oncology ward and an outpatient setting except for blood samplings. The difference is more pronounced between hospital and home care settings, as in the latter the patient performs many of the activities him/herself. The cost estimate per setting for prophylactic drug use is 6.30 Euros for in-hospital care, 3.67 Euros for outpatient care and 5.49 Euros for home care. Taking the two most frequently occurring scenarios per chemotherapy cycle (i.e. with or without febrile neutropenia), the following cost estimates are obtained: 60.41 Euros for a patient with febrile neutropenia and 56.77 Euros for a patient without febrile neutropenia, excluding drug costs. With the activity-based costing method it is possible to accurately demonstrate cost savings in the management of neutropenia using the newer drug therapies. PMID- 15819843 TI - Recent advances in sea-ice microbiology. AB - Over the past 50 years there has been much effort invested in the investigation of the ecology of sea ice. Sea ice is an ephemeral feature of the Arctic and Southern Oceans and smaller water bodies such as the Baltic and Caspian Seas. The semisolid ice matrix provides a range of habitats in which a diverse range of microbial organisms thrive. In the past 5 years there has been considerable steps forward in sea-ice research, in particular regarding the analysis of sea-ice microstructure and the investigation of the diversity and adaptation of microbial communities. These studies include: (i) controlled simulated and in situ studies on a micrometer scale to unravel the dynamic of the microhabitat with consequences for the organisms; (ii) the introduction of molecular approaches to uncover the diversity of uncultured still unknown microorganisms; and (iii) studies into the molecular adaptation of selected model organisms to the extreme environment. This minireview presents some of the most recent findings from sea ice studies within the framework of these aims. PMID- 15819844 TI - Effect of humic material on the bacterioplankton community composition in boreal lakes and mesocosms. AB - The bacterioplankton community composition in two Finnish forest lakes with different content of humic substances was studied by denaturing gradient gel electrophoresis (DGGE) and sequencing of the major bands. The same dominant bacterial phylotypes were detected in the bacterioplankton communities of clear water Lake Ahvenlammi and humic Lake Sammalisto. For 4 years, in every water layer, Actinobacteria was the dominant and Verrucomicrobia the second most common phylum. In the hypolimnion, other dominant phyla were also found. We set up a mesocosm experiment to assess the effect of a sudden load of allochthonous humus extract to the bacterioplankton community composition. Changes in the bacterial communities were followed in four control and four humus extract-added mesocosms for 50 days. In the humic mesocosms the phylotypes of allochthonous Proteobacteria arriving with the humus extract were initially prevalent but disappeared during the first weeks. After this the Actinobacteria-dominated communities resembled the bacterioplankton communities of the control mesocosms and Lake Ahvenlammi. Towards the end of the experiment the community patterns in all the mesocosms started to change slightly because of erratic occurrence of new proteobacterial phylotypes. Thus the effects of a sudden load of allochthonous humic material and bacteria to the bacterioplankton community composition were transient. PMID- 15819845 TI - Phenotypic and genotypic characterization of encapsulated Escherichia coli isolated from blooms in two Australian lakes. AB - Escherichia coli has long been used as an indicator organism for water quality assessment. Recently there has been an accumulation of evidence that suggests some strains of this organism are able to proliferate in the environment, a characteristic that would detract from its utility as an indicator of faecal pollution. Phenotypic and genotypic characterization of E. coli isolated from blooms in two Australian lakes, separated by a distance of approximately 200 km, identified that the blooms were dominated by three E. coli strains. A major phenotypic similarity among the three bloom strains was the presence of a group 1 capsule. Genetic characterization of a conserved region of the cps gene cluster, which encodes group 1 capsules, identified a high degree of genetic variation within the bloom isolates. This differs from previously described encapsulated E. coli strains which are highly conserved at the cps locus. The phenotypic or genotypic profiles of the bloom strains were not identified in 435 E. coli strains isolated from vertebrates. The occurrence of these encapsulated strains suggests that some E. coli have evolved a free-living lifestyle and do not require a host in order to proliferate. The presence of the same three strains in bloom events in different geographical regions of a temperate climate, and at different times, indicates that free-living E. coli strains are able to persist in these water reservoirs. This study provides further evidence of circumstances where caution is required in using E. coli as an indicator organism for water quality. PMID- 15819846 TI - Growth of Geobacter sulfurreducens under nutrient-limiting conditions in continuous culture. AB - A system for growing Geobacter sulfurreducens under anaerobic conditions in chemostats was developed in order to study the physiology of this organism under conditions that might more closely approximate those found in the subsurface than batch cultures. Geobacter sulfurreducens could be cultured under acetate-limiting conditions with fumarate or Fe(III)-citrate as the electron acceptor at growth rates between 0.04 and 0.09 h(-1). The molar growth yield was threefold higher with fumarate as the electron acceptor than with Fe(III), despite the lower mid point potential of the fumarate/succinate redox couple. When growth was limited by availability of fumarate, high steady-state concentrations were detected, suggesting that fumarate is unlikely to be an important electron acceptor in sedimentary environments. The half-saturation constant, Ks, for acetate in Fe(III)-grown cultures (10 microM) suggested that the growth of Geobacter species is likely to be acetate limited in most subsurface sediments, but that when millimolar quantities of acetate are added to the subsurface in order to promote the growth of Geobacter for bioremediation applications, this should be enough to overcome any acetate limitations. When the availability of electron acceptors, rather than acetate, limited growth, G. sulfurreducens was less efficient in incorporating acetate into biomass but had higher respiration rates, a desirable physiological characteristic when adding acetate to stimulate the activity of Geobacter species during in situ uranium bioremediation. These results demonstrate that the ability to study the growth of G. sulfurreducens under steady-state conditions can provide insights into its physiological characteristics that have relevance for its activity in a diversity of sedimentary environments. PMID- 15819847 TI - Genetic analysis of housekeeping genes reveals a deep-sea ecotype of Alteromonas macleodii in the Mediterranean Sea. AB - The genetic diversity of 19 strains belonging to Alteromonas macleodii isolated from different geographic areas (Pacific and Indian Ocean, and different parts of the Mediterranean Sea) and at different depths (from the surface down to 3500 m) has been studied. Fragments of the 16S rRNA gene, the internal transcribed spacer (ITS) between 16S and 23S rDNA genes, the gyrB and the rpoB genes, have been sequenced for each strain. Amplified fragment length polymorphisms were used to characterize similarity at the level of the whole genome. Most of the diversity reflected the existence of a cluster of strains isolated from deep Mediterranean waters and two isolates from the Black Sea. Particularly the isolates from the deep sites were consistently different from all the others indicating the existence of a specific ecotype adapted to these conditions. Amplification of gyrB gene and ITS directly from DNA retrieved from deep Mediterreanean waters and one Atlantic sample showed that presence of this deep-sea ecotype is widespread and is not a product of culture bias. On the other hand, strains isolated from surface tropical waters showed a remarkable level of resemblance to the first isolate of this species obtained from Hawaii in 1972. The results indicate the existence of both lineages of global distribution and ecotypes adapted to specific conditions such as deep or more diluted (the Black Sea) waters. PMID- 15819848 TI - Stereospecific effect of hexachlorocyclohexane on activity and structure of soil methanotrophic communities. AB - In the past decades, large amounts of non-insecticidal hexachlorocyclohexane (HCH) isomers (alpha-, beta-, delta- and epsilon-HCH) have been dumped as side products of the insecticide gamma-HCH (lindane). This study investigates the effect of HCH isomers on methane oxidation, an important soil function performed by methanotrophic bacteria. Both activity and structure of the methanotrophic community were assessed, using methane oxidation assays and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) respectively. Methane oxidation assays with historically polluted soils revealed that on the long-term methane oxidation was inhibited by HCH pollution. PCR-DGGE and diversity analysis based on Lorenz curves showed that the type I methanotrophic community was less evenly distributed in historically HCH-polluted soils compared with less polluted reference soils. Short-term experiments with methane-enriched consortia further demonstrated that only gamma- and delta-isomers inhibited methane oxidation. Type I methanotrophs of methane-enriched microbial consortia that received gamma- or delta-HCH evolved towards higher species richness. Apparently, for historically HCH-polluted soils, a narrow community remained after long-term exposure while in case of short-term exposures, methane-enriched consortia were converted into less active, but richer communities when they were stressed by the presence of gamma- or delta-HCH. This work demonstrates the importance of incorporating all isomers and possible other side-products in risk assessment studies of persistent organic pollutants and the use of structural analysis of type I methanotrophic communities as evaluating tool. PMID- 15819849 TI - Retrieval of nearly complete 16S rRNA gene sequences from environmental DNA following 16S rRNA-based community fingerprinting. AB - 16S rRNA-based fingerprinting techniques allow rapid analyses of overall bacterial community structure but suffer from a lack of phylogenetic information hitherto retrievable from the short 16S rRNA gene sequences obtained from excised bands. An approach is presented that allows nearly complete 16S rRNA gene sequences to be retrieved for abundant components of the bacterial community as obtained by the community fingerprint, i.e. those reflected by major fingerprint bands. This was achieved by designing a pair of highly specific primers derived from the sequence of an excised band. Combined with universal 16S rRNA primers, these specific primers were applied directly to environmental DNA serving as template. This procedure allowed the generation of a nearly complete 16S rRNA gene sequence of the target taxon by specific polymerase chain reaction (PCR) followed by cycle sequencing down to a relative abundance of at least 1.5% of the environmental DNA. The procedure was exemplified for an epsilonproteobacterium related to Thiomicrospira denitrificans occurring in the central Baltic Sea. This approach is based only on PCR without any cloning step involved. It allows focussing on specific target taxa and is thus rather efficient. This approach should be applicable in general to 16S rRNA or 16S rRNA gene-based fingerprinting techniques and their respective environmental DNA. PMID- 15819850 TI - Links between ammonia oxidizer species composition, functional diversity and nitrification kinetics in grassland soils. AB - Molecular approaches have revealed considerable diversity and uncultured novelty in natural prokaryotic populations, but not direct links between the new genotypes detected and ecosystem processes. Here we describe the influence of the structure of communities of ammonia-oxidizing bacteria on nitrogen cycling in microcosms containing natural and managed grasslands and amended with artificial sheep urine, a major factor determining local ammonia concentrations in these environments. Nitrification kinetics were assessed by analysis of changes in urea, ammonia, nitrite and nitrate concentrations and ammonia oxidizer communities were characterized by analysis of 16S rRNA genes amplified from extracted DNA using ammonia oxidizer-specific primers. In natural soils, ammonia oxidizer community structure determined the delay preceding nitrification, which depended on the relative abundance of two Nitrosospira clusters, termed 3a and 3b. In batch cultures, pure culture and enrichment culture representatives of Nitrosospira 3a were sensitive to high ammonia concentration, while Nitrosospira cluster 3b representatives and Nitrosomonas europaea were tolerant. Delays in nitrification occurred in natural soils dominated by Nitrosospira cluster 3a and resulted from the time required for growth of low concentrations of Nitrosospira cluster 3b. In microcosms dominated by Nitrosospira cluster 3b and Nitrosomonas, no substantial delays were observed. In managed soils, no delays in nitrification were detected, regardless of initial ammonia oxidizer community structure, most probably resulting from higher ammonia oxidizer cell concentrations. The data therefore demonstrate a direct link between bacterial community structure, physiological diversity and ecosystem function. PMID- 15819851 TI - High diversity of the 'Spumella-like' flagellates: an investigation based on the SSU rRNA gene sequences of isolates from habitats located in six different geographic regions. AB - We isolated 28 strains of 'Spumella-like' flagellates from different freshwater and soil habitats in Austria, People's Republic of China, Nepal, New Zealand, Uganda, Kenya, Tanzania and Hawaii by use of a modified filtration acclimatization method. 'Spumella-like' flagellates were found in all of the samples and were often among the dominant bacterivorous flagellates in the respective environments. The small subunit ribosomal RNA (SSU rRNA) gene sequence of the isolates was determined and aligned with previously published sequences of members belonging to the Chrysophyceae sensu stricto. Phylogenetic analysis of the 28 new sequences confirmed their position within the Chrysophyceae sensu stricto and positioned them within different clades. Most of the sequences grouped within clade C and formed several subclusters separated from each other by green taxa including flagellates belonging to Ochromonas, Dinobryon, Poterioochromonas and others. All soil isolates clustered together (subcluster C1) with the soil strain Spumella elongata and the undescribed soil strain 'Spumella danica'. Aquatic isolates were affiliated with at least two branches (C2 and C3). Sequence similarity to the closest related member of the Chrysophyceae ranged between 92% and 99.6%, sequence divergence among the 'Spumella-like' flagellates was as high as 10%. We conclude that (i) the 'Spumella-like' flagellates are a diverse group both in terms of sequence dissimilarity between isolates and in terms of the number of genotypes, (ii) Spumella and Ochromonas are polyphyletic, and (iii) based on the SSU rRNA gene no biogeographical restriction of certain branches could be observed even though different ecotypes may be represented by the same genotype. PMID- 15819852 TI - Diversity of Bacteria and Archaea associated with a carbonate-rich metalliferous sediment sample from the Rainbow vent field on the Mid-Atlantic Ridge. AB - Two sediment cores were collected in an inactive area of the deep-sea hydrothermal vent field Rainbow (36 degrees N on the Mid-Atlantic Ridge). Metals and carbonates were abundant throughout the cores; calcite (CaCO3) was found throughout the cores while dolomite [CaMg(CO3)2] and siderite (FeCO3) were only found in deeper layers. Using polymerase chain reaction (PCR)-amplified 16S rRNA gene sequence analysis, we examined the bacterial and archaeal diversity in a sediment layer that contained the three carbonates. The retrieved bacterial and archaeal sequences were new and less than 4% of the sequences exhibited 94% or more identity with that of cultured organisms. The analysis of the composition of the bacterial library revealed a high diversity of sequences. Half of the bacterial clones was affiliated to the gamma-Proteobacteria. Most of them had environmental sequences retrieved from deep-sea sediments as closest relatives, some of which being distantly related to free-living and symbiotic sulfur oxidizers. Other sequences clustered in the alpha-, delta- and epsilon Proteobacteria, the 'Bacteroidetes', the 'Planctomycetes', the 'Nitrospirae', the 'Actinobacteria', the 'Chlorobi ' and the 'Verrumicrobia'. Based on clonal abundance and sequence comparisons, phylotype groups putatively involved in the oxydation of sulfur compounds appeared to dominate in the studied sample. The majority of the archaeal sequences clustered in an euryarchaeotic lineage recently identified in the walls of black smokers suggesting a possible thermophilic way of life of these uncultured microorganisms. Oxygen isotopic composition of siderite and dolomite indicated that they were formed at 67 degrees C and 94 degrees C respectively. Together with chemical and microbiological data, this suggested that hydrothermal fluids may have circulated through this sediment. PMID- 15819853 TI - Functional and compositional comparison of two activated sludge communities remediating coking effluent. AB - The success of engineered microbiological systems is evident in the global application of activated sludge communities to remediate coking effluent. However, there is a lack of understanding of the microbiology underlying treatment efficiency and stability. In this study, two functionally distinct activated sludge pools, treating the same effluent and operating under the same conditions, were examined to establish a relationship between overall diversity and/or functional diversity with respect to process stability. Molecular profiling, sequencing and RNA-based stable isotope probing were used to examine the bacterial diversity, general composition and functional composition of the most abundant members of the two communities. The inferior process stability in one of the pools could not be explained by reduced total bacterial diversity or evenness. RNA-based stable isotope probing revealed that both pools harboured an abundant phenol-degrading Acidovorax species, and that the pool of inferior stability accommodated an additional closely related phenol-degrading Acidovorax species at high abundance. These results are discussed in the context of deterministic and stochastic models of microbial community assembly. PMID- 15819854 TI - Bacterial diversity in the bacterioneuston (sea surface microlayer): the bacterioneuston through the looking glass. AB - The bacterioneuston is defined as the community of bacteria present within the neuston or sea surface microlayer. Bacteria within this layer were sampled using a membrane filter technique and bacterial diversity was compared with that in the underlying pelagic coastal seawater using molecular ecological techniques. 16S rRNA gene libraries of approximately 500 clones were constructed from both bacterioneuston and the pelagic water samples and representative clones from each library were sequenced for comparison of bacterial diversity. The bacterioneuston was found to have a significantly lower bacterial diversity than the pelagic seawater, with only nine clone types (ecotaxa) as opposed to 46 ecotaxa in the pelagic seawater library. Surprisingly, the bacterioneuston clone library was dominated by 16S rRNA gene sequences affiliated to two groups of organisms, Vibrio spp. which accounted for over 68% of clones and Pseudoalteromonas spp. accounting for 21% of the library. The dominance of these two 16S rRNA gene sequence types within the bacterioneuston clone library was confirmed in a subsequent gene probing experiment. 16S rRNA gene probes specific for these groups of bacteria were designed and used to probe new libraries of 1000 clones from both the bacterioneuston and pelagic seawater DNA samples. This revealed that 57% of clones from the bacterioneuston library hybridized to a Vibrio sp. specific 16S rRNA gene probe and 32% hybridized to a Pseudoalteromonas sp. specific 16S rRNA gene probe. In contrast, the pelagic seawater library resulted in only 13% and 8% of 16S rRNA gene clones hybridizing to the Vibrio sp. and Pseudoalteromonas sp. probes respectively. Results from this study suggest that the bacterioneuston contains a distinct population of bacteria and warrants further detailed study at the molecular level. PMID- 15819855 TI - Use of monoclonal antibodies to quantify the dynamics of alpha-galactosidase and endo-1,4-beta-glucanase production by Trichoderma hamatum during saprotrophic growth and sporulation in peat. AB - Trichoderma species are ubiquitous soil and peat-borne saprotrophs that have received enormous scientific interest as biocontrol agents of plant diseases caused by destructive root pathogens. Mechanisms of biocontrol such as antibiosis and hyperparasitism are well documented and the biochemistry and molecular genetics of these processes defined. An aspect of biocontrol that has received little attention is the ability of Trichoderma species to compete for nutrients in their natural environments. Trichoderma species are efficient producers of polysaccharide-degrading enzymes that enable them to colonize organic matter thereby preventing the saprotrophic spread of plant pathogens. This study details the use of monoclonal antibodies (mAbs) to quantify the production of two enzymes implicated in the saprotrophic growth of Trichoderma species in peat. Using mAbs specific to the hemicellulase enzyme alpha-galactosidase (AGL) and the cellulase enzyme endo-1,4-beta-glucanase (EG), the relationship between the saprotrophic growth dynamics of a biocontrol strain of Trichoderma hamatum and the concomitant production of these enzymes in peat-based microcosms was studied. Enzyme activity assays and enzyme protein concentrations derived by enzyme-linked immunosorbent assay (ELISA) established the precision and sensitivity of mAb-based assays in quantifying enzyme production during active growth of the fungus. Trends in enzyme activities and protein concentrations were similar for both enzymes, during a 21-day sampling period in which active growth and sporulation of the fungus in peat was quantified using an independent mAb-based assay. There was a sharp increase in active biomass of T. hamatum 3 days after inoculation of microcosms with phialoconidia. After 3 days there was a rapid decline in active biomass which coincided with sporulation of the fungus. A similar trend was witnessed with EG activities and concentrations. This showed that EG production related directly to active growth of the fungus. The trend was not found, however, with AGL. There was a rapid increase in enzyme activities and protein concentrations on day 3, after which they remained static. The reason for the maintenance of elevated AGL probably resulted from secretion of the enzyme from conidia and chlamydospores. ELISA, immunofluoresence and immunogold electron microscopy studies of these cells showed that the enzyme is localized within the cytoplasm and is secreted extracellularly into the surrounding environment. It is postulated that release of oligosaccharides from polymeric hemicellulose by the constitutive spore-bound enzyme leads to AGL induction and could act as an environmental cue for spore germination. PMID- 15819856 TI - Hydrothermal vent gastropods from the same family (Provannidae) harbour epsilon- and gamma-proteobacterial endosymbionts. AB - The discovery of new hydrothermal vent systems in the back-arc basins of the Western Pacific revealed chemosynthesis-based faunal communities distinct from those of other vents. These vents are dominated by two related gastropods (Alviniconcha spp. and Ifremeria nautilei) that harbour symbiotic bacteria in their gills. We used comparative 16S ribosomal RNA (rRNA) gene sequencing and in situ hybridization with rRNA-targeted probes to characterize the bacterial symbionts of Alviniconcha sp. and I. nautilei from the Manus Basin in the Western Pacific. The analyses revealed that these two gastropod species, although affiliated with the same family, harbour phylogenetically distant chemosymbionts, suggesting independent origins of these endosymbioses. The I. nautilei endosymbiont clusters with sulfur-oxidizing bacteria within the gamma Proteobacteria, as is the case for all previously characterized endosymbionts from a wide diversity of host taxa harbouring thioautotrophic prokaryotes. In contrast, the Alviniconcha endosymbiont is affiliated with sulfur-oxidizing bacteria within the epsilon-Proteobacteria. These results show that bacteria from the epsilon-Proteobacteria are also capable of forming endosymbiotic associations with marine invertebrates from chemosynthetic environments. More generally, the endosymbiotic lifestyle is now shown to be distributed throughout all recognized classes of the Proteobacteria. PMID- 15819858 TI - Cutaneous leishmaniasis: an increasing threat for travellers. AB - Analysis of the literature on cutaneous leishmaniasis in low-prevalence countries suggests an increase in imported cases that is attributable to the growing phenomenon of international tourism, migration and military operations in highly endemic regions. Cases of imported cutaneous leishmaniasis are often missed initially, but diagnosis can be made non-invasively by PCR using skin scrapings of lesions as starting material. Cutaneous leishmaniasis is an emerging threat for travellers and should be considered in all patients presenting with slow-to heal ulcers. PMID- 15819859 TI - Shewanella algae and Shewanella putrefaciens: clinical and microbiological characteristics. AB - The occurrence of the two Shewanella species found in clinical specimens, Shewanella algae and Shewanella putrefaciens, correlates with the temperature and salinity of seawater. This means that Shewanella infections occur in warm climates or during especially warm summers in temperate climates. The infections described most commonly involve ears, skin and soft tissue, with or without bacteraemia. Primary bacteraemia with a fulminant course is also seen in immunocompromised patients. Important differential characteristics between the two species include the ability of S. algae to produce mucoid colonies with beta haemolysis on sheep blood agar, to grow at 42 degrees C and in NaCl 6% w/v, and to reduce nitrite, and an inability to produce acid from maltose, all of which are in contrast to the characteristics of S. putrefaciens. Automated identification systems fail to differentiate between S. algae and S. putrefaciens, as S. algae is not included in the databases of these systems. Presumably for this reason, most Shewanella infections reported during recent years have been attributed to S. putrefaciens. However, when extensive phenotypic characterisation is performed, most human infections are seen to be caused by S. algae. As the two species seem to have different pathogenic potential for humans, correct identification is important, and this is possible in routine clinical microbiology laboratories. PMID- 15819860 TI - Rapid identification of Candida spp. in peritonitis patients by Raman spectroscopy. AB - This prospective study evaluated Raman spectroscopy for the identification of clinically relevant Candida spp. in peritonitis patients. A Raman database was developed by measuring spectra from 93 reference strains belonging to ten different Candida spp. Clinical samples were obtained from the surgical department and intensive care unit of a tertiary university hospital. In total, 88 peritoneal specimens from 45 patients with primary, secondary or tertiary peritonitis were included. Specimens were cultured initially on a selective Sabouraud medium that contained gentamicin to suppress bacterial growth. For conventional identification, a chromogenic medium was used for presumptive identification, followed by use of the Vitek 2 system for definitive identification (requiring a total time of 48-96 h). Raman measurements were taken on overnight cultures from Sabouraud-gentamicin medium. Thirty-one samples were positive for Candida by culture. Using multivariate statistical analyses, a prediction accuracy of 90% was obtained for Raman spectroscopy, which appears to offer an accurate and rapid (12-24 h) alternative for the identification of Candida spp. in peritonitis patients. The reduced turn-around time is of great clinical importance for the treatment of critically ill patients with invasive candidiasis in intensive care units. PMID- 15819861 TI - Aspergillus infections in lung transplant recipients: risk factors and outcome. AB - This retrospective study of 251 lung transplant patients aimed to determine the prevalence, clinical presentation and mortality of Aspergillus infection in order to define specific risk factors and to compare survival in patients with and without infection. Aspergillus was isolated from 86 (33%) cases, which involved colonisation (n = 50), tracheobronchial lesions (n = 17) or invasive aspergillosis (n = 19). Overall, aspergillosis had an impact on survival (p < 0.05); in fact the 5-year mortality rate was substantially higher in single lung transplant recipients with bronchial anastomotic infection, and in those with late-onset infections and chronic rejection. A significant association (p < 0.05) was found between acute rejection and the time at which fungal infection was diagnosed. Aspergillus infection was not related to cytomegalovirus infection or treatment with corticosteroids. The mortality rate for invasive infections was 78% and was related to survival (p < 0.0001); invasive aspergillosis was also associated with chronic rejection (p < 0.05), but not with high corticosteroid doses (p 0.49) or use of tacrolimus (p 0.73). In conclusion, Aspergillus infection was associated with a reduction in the 5-year survival rate of lung transplant recipients, and this was particularly true for patients infected with the invasive forms and for patients with single lung transplants, bronchial anastomotic infection and chronic rejection. Isolation of Aspergillus spp. from respiratory samples preceded acute rejection, and may be a marker of graft dysfunction and/or airway inflammation. Close monitoring, or even pre-emptive antifungal therapy, is recommended for patients with chronic rejection or bronchial airway mechanical abnormalities and persistent Aspergillus colonisation. PMID- 15819862 TI - Human metapneumovirus-associated respiratory tract infections in the Republic of Ireland during the influenza season of 2003-2004. AB - Human metapneumovirus (hMPV) is a newly identified paramyxovirus that has been associated with respiratory tract illness in children aged < 5 years, the elderly, and immunocompromised patients. This study determined the frequency of respiratory tract infections (RTIs) associated with hMPV in the Republic of Ireland. Bronchoalveolar lavage (BAL) samples from 168 adult patients and respiratory specimens from 122 children aged < 5 years were collected between September 2003 and May 2004. The virus was detected by reverse-transcription (RT) PCR using hMPV polymerase (L) and matrix (M)-specific primers in four (2.4%) of 171 BAL specimens obtained from 168 adults. No other respiratory virus was detected in these specimens, and no hMPV RNA was detected in respiratory specimens from children during the same time period. In all four adult cases, two of whom had underlying disease, hMPV was associated with mild, self-limiting upper RTIs. The most common clinical findings included fever (3/4 patients), cough (4/4) and rhinorrhoea (3/4). No patient died as a result of these RTI episodes. Phylogenetic analysis was performed using the amplified regions of the M and fusion (F) genes of hMPV. The Irish isolates belonged to cluster 1B, and did not show a separate Irish sub-lineage. PMID- 15819863 TI - Susceptibility testing of Mycobacterium tuberculosis: comparison of the BACTEC TB 460 method and flow cytometric assay with the proportion method. AB - Tuberculosis is a leading cause of morbidity and mortality worldwide. Susceptibility testing of the causative agent, Mycobacterium tuberculosis, is critical for control of the disease. This study compared the flow cytometric susceptibility assay with the proportion method and the BACTEC TB-460 system. There was agreement between the flow cytometric and proportion methods for 73 (94%) of 78 isoniazid tests, and complete agreement for 26 ethambutol and rifampicin tests. In contrast, the proportion and BACTEC methods failed to agree for 22%, 15% and 8% of isoniazid, ethambutol and rifampicin tests, respectively. These findings indicated that susceptibility testing by the flow cytometric assay is accurate, with results available within 24 h of initiation of the testing procedure. PMID- 15819864 TI - Antimicrobial susceptibility patterns of Staphylococcus aureus in Poland obtained by the National Quality Assurance Programme. AB - As part of the Polish external quality assurance scheme, clinical laboratories were asked to send five consecutive isolates of Staphylococcus aureus and the corresponding susceptibility results to the national Centre of Quality Control in Microbiology. Of 1376 isolates submitted as S. aureus from 276 medical centres, 13 (< 1%) had been misidentified by local laboratories. Of 181 (13.5%) methicillin-resistant S. aureus (MRSA) isolates, most were identified correctly (c. 98% of laboratories). Although all MRSA isolates were fully susceptible to vancomycin, teicoplanin and linezolid, they were usually multiresistant; almost 23% were resistant to seven antimicrobial agents. Most (> 90%) MSSA isolates were susceptible to the tested antibiotics, except penicillin (21% susceptible) and tetracycline (62.4% susceptible). In addition to evaluating the proficiency of testing by local laboratories, the study yielded valuable information regarding the susceptibility patterns of S. aureus isolates in Poland. PMID- 15819865 TI - Simultaneous single-tube PCR assay for the detection of Neisseria meningitidis, Haemophilus influenzae type b and Streptococcus pneumoniae. AB - Rapid, accurate and inexpensive diagnosis of bacterial meningitis is critical for patient management. This study describes the development and evaluation of a multiplex PCR assay for the detection of Neisseria meningitidis, Streptococcus pneumoniae and Haemophilus influenzae type b, which globally account for 90% of cases of bacterial meningitis. The single-tube assay, based on the ctrA, ply and bex targets, respectively, enabled detection of 5-10 pg DNA. When the assay was tested with clinical samples (n = 425), its sensitivity for the three targets was 93.9%, 92.3% and 88%, respectively, while the overall specificity and positive predictive value of the assay was 100%. The negative predictive value was 99.1 99.5%. The methodology permits rapid and accurate detection of the three main pathogens that cause bacterial meningitis. PMID- 15819866 TI - Clinical presentation of leptospirosis: a retrospective study of 34 patients admitted to a single institution in metropolitan France. AB - Leptospirosis has a highly variable clinical presentation, which may be related to different infecting serovars, host factors, or a combination of these. This study investigated retrospectively 34 consecutive patients with serologically confirmed leptospirosis admitted during the period 1992-2002. On admission, the most frequent symptoms were fever (100%), headache (75%), myalgia (55%), arthralgia (45%) and vomiting (39%). Pertinent laboratory findings included lymphopenia (85%), thrombocytopenia (75%), elevated liver enzymes (87%) and renal abnormalities (proteinuria, 77%; haematuria, 58%; elevated serum creatinine, 53%). The study confirmed the variable clinical and biological symptoms of leptospirosis, and indicated that lymphopenia is a common feature of leptospirosis cases. PMID- 15819867 TI - Molecular diversity of Proteus mirabilis isolates producing extended-spectrum beta-lactamases in a French university hospital. AB - Between February 1997 and December 2002, 3340 hospitalised patients yielded samples positive for Proteus mirabilis, of whom 45 (1.3%) were colonised/infected by P. mirabilis producing extended-spectrum beta-lactamases (ESBLs). The gross incidence of patients colonised/infected by ESBL-producing P. mirabilis was 1.61/10(5) days of hospitalisation, with 20% of isolates being collected from patients in urology wards, most frequently (53.3%) from urine samples. Seventeen (37.7%) of the 43 isolates were obtained from samples collected within 48 h of hospitalisation, indicating that they were community-acquired. Isoelectric focusing assays and sequencing identified the TEM-24, TEM-92 and TEM-52 ESBLs. Pulsed-field gel electrophoresis revealed eight pulsotypes (I-VIII), with the two most common pulsotypes, IV and VI, comprising ten (23.3%) and 12 (26.6%) isolates, respectively. These pulsotypes were considered to represent epidemic strains and spread in various wards of the hospital. PMID- 15819869 TI - Incidence of glycopeptide hetero-intermediate Staphylococcus aureus strains in Maltese hospitals. AB - The incidence of hetero-intermediate glycopeptide susceptibility among Staphylococcus aureus isolates in Malta, a country with a high incidence of methicillin resistance, was studied by screening 454 non-repetitive S. aureus isolates on teicoplanin-supplemented agar plates, followed by Etests and genotypic studies. All strains were susceptible to vancomycin, but four (0.88%) exhibited teicoplanin MICs of > 12 mg/L. High methicillin-resistant S.aureus endemicity was not an accurate predictor of the emergence of non-susceptibility to glycopeptides. PMID- 15819868 TI - Evaluation of antimicrobial susceptibility of bacteria containing the qnr gene and FOX-5 beta-lactamase by four automated systems. AB - The accuracy and performance of four automated instruments (BD Phoenix, MicroScan WalkAway, VITEK-2 and Wider) were evaluated for susceptibility testing of fluoroquinolones and beta-lactams with four clinical isolates of Klebsiella pneumoniae and the corresponding Escherichia coli transconjugants containing a plasmid carrying the qnr gene and coding for FOX-5 production. No major or very major errors were detected with the MicroScan system. Many of the minor errors for both quinolones and beta-lactams clustered around the intermediate breakpoints. PMID- 15819870 TI - Microbiology of sinusitis and the predictive value of throat culture for the aetiology of sinusitis. AB - A prospective study of throat cultures and maxillary sinus aspirates from children with chronic sinusitis (n = 21), acute sinusitis (n = 28) or a clinical diagnosis of chronic adenoiditis (n = 41) was performed. Seventy-two bacterial pathogens were isolated from sinus aspirates from 52% of the study population. Haemophilus influenzae was most common pathogen, followed by Moraxella catarrhalis, Streptococcus pneumoniae, Staphylococcus aureus, and group A streptococci. Quantitative throat cultures had positive predictive values of 41%, 53% and 75% for H. influenzae, Strep. pneumoniae and M. catarrhalis, respectively, while negative predictive values were 93-98%, indicating that these three pathogens do not cause sinusitis when absent from the throat. PMID- 15819871 TI - No evidence of Legionella infection in general practice patients presenting with acute respiratory infections in The Netherlands. AB - The role of Legionella spp. in the aetiology of acute respiratory infections (ARIs) is largely unknown. In this case-control study, conducted in a general practitioner setting during 2000 and 2001, nose and throat samples from patients presenting with ARIs (n = 230) and controls (n = 200) were analysed for the presence of Legionella spp. by real-time PCR. Legionella DNA was not detected in any of the cases or controls. Thus, Legionella spp. do not seem to play a role in patients presenting with ARIs, nor were they present in patients who visited their general practitioner for complaints other than ARIs. PMID- 15819872 TI - Surveillance of Legionnaires' disease in Austria. AB - Seven foreign cases of travel-associated Legionnaires' disease were associated with two neighbouring tourist villages in Austria between 1992 and 2004. The seven foreigners stayed in four hotels located in a geographical diameter of less than 10 km. Two cases were diagnosed in The Netherlands and five in the UK. The first case occurred in 1992, while the remaining cases clustered between August 2000 and April 2004. None of these cases was reported officially as part of a hotel-associated cluster, thereby highlighting several weaknesses of the infectious disease control systems involved. PMID- 15819873 TI - Clinical features and outcome of patients with community-acquired Pseudomonas aeruginosa bacteraemia. AB - Cases of community-acquired Pseudomonas aeruginosa bacteraemia (n = 39) that occurred at a tertiary-care hospital during a 5-year period were analysed retrospectively. The commonest underlying diseases were solid tumour (41%) and haematological malignancy (18%). Most (44%) of the patients were neutropenic, and 39% had septic shock at initial presentation. The 30-day attributable mortality rate was 39%. Two previously healthy patients were identified with fatal P. aeruginosa pneumonia with bacteraemia. P. aeruginosa bacteraemia is a fatal infection that should be considered in the differential diagnosis of patients presenting from the community with rapidly progressive sepsis. PMID- 15819874 TI - Continuous infusion beta-lactams for intensive care unit pulmonary infections. AB - This study evaluated the pharmacodynamics of continuous infusion beta-lactams against pulmonary isolates of Gram-negative bacteria from patients managed in intensive care units (ICUs) in the USA. Multiple 10,000-patient Monte Carlo simulations were performed by integrating pharmacokinetic data from healthy individuals with 2408 MICs from the 2002 Intensive Care Unit Surveillance System database. These pharmacodynamic simulations suggested that continuous infusion regimens of cefepime, aztreonam, ceftazidime and piperacillin-tazobactam 13.5 g have the greatest likelihood of achieving pharmacodynamic targets against isolates of Enterobacteriaceae in the ICU. Beta-lactams are unlikely to achieve pharmacodynamic targets against Pseudomonas aeruginosa or Acinetobacter baumannii when administered as monotherapy. PMID- 15819875 TI - Both penicillin and amoxycillin should be tested in antimicrobial surveillance for Streptococcus pneumoniae. PMID- 15819876 TI - Performance of the Focus ELISA test for detection of herpes simplex virus type-2 specific antibodies in Chinese STD patients. PMID- 15819877 TI - Immunisation against tick-borne encephalitis by widely used vaccines: short-term history and current recommendations. PMID- 15819878 TI - Short hydrogen bonds in proteins. AB - Short hydrogen bonds are present in many chemical and biological systems. It is well known that these short hydrogen bonds are found in the active site of enzymes and aid enzyme catalysis. This study aims to systematically characterize all short hydrogen bonds from a nonredundant dataset of protein structures. The study has revealed that short hydrogen bonds are commonly found in proteins and are widely present in different regions of the protein chain, such as the backbone or side chain, and in different secondary structural regions such as helices, strands and turns. The frequency of occurrence of donors and acceptors from the charged side chains as well as from the neutral backbone atoms is equally high. This suggests that short hydrogen bonds in proteins occur either due to increased strength or due to geometrical constraints and this has been illustrated from several examples. PMID- 15819879 TI - Adhesion properties of adhesion-regulating molecule 1 protein on endothelial cells. AB - Numerous adhesion molecules have been described, and the molecular mechanisms of lymphocyte trafficking across the endothelium is starting to be elucidated. Identification of the molecules involved in the organoselectivity of this process would help in the targeting of drug therapy to specific tissues. Adhesion regulating molecule-1 (ARM-1) is an adhesion-regulating molecule previously identified on T cells. It does not belong to any known families of adhesion molecules. In this study, we show the presence of ARM-1 in endothelial cells, the adhesion partners of lymphocytes. ARM-1 mRNA was found to be differentially expressed in endothelial cell lines of various tissue origin and lymphocyte cell lines. Interestingly, ARM-1 is absent from skin endothelial cells. In our assay, skin endothelial cells display a distinct capacity to mediate adhesion of activated T lymphocytes. Overexpression of ARM-1 in skin endothelial cells increased adhesion of CEMT4 and NK lymphocytes, confirming that ARM-1 also regulates adhesion in endothelial cells. We also show that ARM-1 is a cytosolic protein associated with the plasma membrane. However, no cell surface expression of the protein was observed. These results suggest an indirect role of ARM-1 in adhesion rather than a direct role as an adhesion molecule itself. PMID- 15819880 TI - 14-3-3 proteins regulate glycogen synthase 3beta phosphorylation and inhibit cardiomyocyte hypertrophy. AB - 14-3-3 proteins are dimeric phophoserine-binding molecules that participate in important cellular processes such as cell proliferation, cell-cycle control and the stress response. In this work, we report that several isoforms of 14-3-3s are expressed in neonatal rat cardiomyocytes. To understand their function, we utilized a general 14-3-3 peptide inhibitor, R18, to disrupt 14-3-3 functions in cardiomyocytes. Cardiomyocytes infected with adenovirus-expressing YFP-R18 (AdR18) exhibited markedly increased protein synthesis and atrial natriuretic peptide production and potentiated the responses to norepinephrine stimulation. This response was blocked by the pretreatment with LY294002, a phosphoinositide 3 kinase (PI3K) inhibitor. Consistent with a role of PI3K in the R18 effect, R18 induced phosphorylation of a protein cloned from the vakt oncogene of retrovirus AKT8 (Akt - also called protein kinase B, PKB) at Ser473 and glycogen synthase 3beta (GSK3beta) at Ser9, but not extracellular signal-regulated kinase 1/2 (ERK1/2). AdR18-induced PKB and GSK3beta phosphorylation was completely blocked by LY294002. In addition, a member of the nuclear factor of activated T cells (NFAT) family, NFAT3, was converted into faster mobility forms and translocated into the nucleus upon the treatment of AdR18. These results suggest that 14-3-3s inhibits cardiomyocytes hypertrophy through regulation of the PI3K/PKB/GSK3beta and NFAT pathway. PMID- 15819881 TI - Light-induced reactions of Escherichia coli DNA photolyase monitored by Fourier transform infrared spectroscopy. AB - Cyclobutane-type pyrimidine dimers generated by ultraviolet irradiation of DNA can be cleaved by DNA photolyase. The enzyme-catalysed reaction is believed to be initiated by the light-induced transfer of an electron from the anionic FADH- chromophore of the enzyme to the pyrimidine dimer. In this contribution, first infrared experiments using a novel E109A mutant of Escherichia coli DNA photolyase, which is catalytically active but unable to bind the second cofactor methenyltetrahydrofolate, are described. A stable blue-coloured form of the enzyme carrying a neutral FADH radical cofactor can be interpreted as an intermediate analogue of the light-driven DNA repair reaction and can be reduced to the enzymatically active FADH- form by red-light irradiation. Difference Fourier transform infrared (FT-IR) spectroscopy was used to monitor vibronic bands of the blue radical form and of the fully reduced FADH- form of the enzyme. Preliminary band assignments are based on experiments with 15N-labelled enzyme and on experiments with D2O as solvent. Difference FT-IR measurements were also used to observe the formation of thymidine dimers by ultraviolet irradiation and their repair by light-driven photolyase catalysis. This study provides the basis for future time-resolved FT-IR studies which are aimed at an elucidation of a detailed molecular picture of the light-driven DNA repair process. PMID- 15819882 TI - Correlation between conformational stability of the ternary enzyme-substrate complex and domain closure of 3-phosphoglycerate kinase. AB - 3-phosphoglycerate kinase (PGK) is a typical two-domain hinge-bending enzyme with a well-structured interdomain region. The mechanism of domain-domain interaction and its regulation by substrate binding is not yet fully understood. Here the existence of strong cooperativity between the two domains was demonstrated by following heat transitions of pig muscle and yeast PGKs using differential scanning microcalorimetry and fluorimetry. Two mutants of yeast PGK containing a single tryptophan fluorophore either in the N- or in the C-terminal domain were also studied. The coincidence of the calorimetric and fluorimetric heat transitions in all cases indicated simultaneous, highly cooperative unfolding of the two domains. This cooperativity is preserved in the presence of substrates: 3 phosphoglycerate bound to the N domain or the nucleotide (MgADP, MgATP) bound to the C domain increased the structural stability of the whole molecule. A structural explanation of domain-domain interaction is suggested by analysis of the atomic contacts in 12 different PGK crystal structures. Well-defined backbone and side-chain H bonds, and hydrophobic and electrostatic interactions between side chains of conserved residues are proposed to be responsible for domain domain communication. Upon binding of each substrate newly formed molecular contacts are identified that firstly explain the order of the increased heat stability in the various binary complexes, and secondly describe the possible route of transmission of the substrate-induced conformational effects from one domain to the other. The largest stability is characteristic of the native ternary complex and is abolished in the case of a chemically modified inactive form of PGK, the domain closure of which was previously shown to be prevented [Sinev MA, Razgulyaev OI, Vas M, Timchenko AA & Ptitsyn OB (1989) Eur J Biochem180, 61-66]. Thus, conformational stability correlates with domain closure that requires simultaneous binding of both substrates. PMID- 15819883 TI - A novel hyperthermostable 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Sulfolobus solfataricus. AB - We report herein the first molecular characterization of 5'-deoxy-5'-methylthio adenosine phosphorylase II from Sulfolobus solfataricus (SsMTAPII). The isolated gene of SsMTAPII was overexpressed in Escherichia coli BL21. Purified recombinant SsMTAPII is a homohexamer of 180 kDa with an extremely low Km (0.7 microm) for 5' deoxy-5'-methylthioadenosine. The enzyme is highly thermophilic with an optimum temperature of 120 degrees C and extremely thermostable with an apparent Tm of 112 degrees C that increases in the presence of substrates. The enzyme is characterized by high kinetic stability and remarkable SDS resistance and is also resistant to guanidinium chloride-induced unfolding with a transition midpoint of 3.3 m after 22-h incubation. Limited proteolysis experiments indicated that the only one proteolytic cleavage site is localized in the C-terminal region and that the C-terminal peptide is necessary for the integrity of the active site. Moreover, the binding of 5'-deoxy-5'-methylthioadenosine induces a conformational transition that protected the enzyme against protease inactivation. By site directed mutagenesis we demonstrated that Cys259, Cys261 and Cys262 play an important role in the enzyme stability since the mutants C259S/C261S and C262S show thermophilicity and thermostability features significantly lower than those of the wild-type enzyme. In order to get insight into the physiological role of SsMTAPII a comparative kinetic analysis with the homologous 5'-deoxy-5' methylthioadenosine phosphorylase from Sulfolobus solfataricus (SsMTAP) was carried out. Finally, the alignment of the protein sequence of SsMTAPII with those of SsMTAP and human 5'-deoxy-5'-methylthioadenosine phosphorylase (hMTAP) shows several key residue changes that may account why SsMTAPII, unlike hMTAP, is able to recognize adenosine as substrate. PMID- 15819884 TI - Plasmodium falciparum hypoxanthine guanine phosphoribosyltransferase. Stability studies on the product-activated enzyme. AB - Hypoxanthine guanine phosphoribosyltransferases (HGPRTs) catalyze the conversion of 6-oxopurine bases to their respective nucleotides, the phosphoribosyl group being derived from phosphoribosyl pyrophosphate. Recombinant Plasmodium falciparum HGPRT, on purification, has negligible activity, and previous reports have shown that high activities can be achieved upon incubation of recombinant enzyme with the substrates hypoxanthine and phosphoribosyl pyrophosphate [Keough DT, Ng AL, Winzor DJ, Emmerson BT & de Jersey J (1999) Mol Biochem Parasitol98, 29-41; Sujay Subbayya IN & Balaram H (2000) Biochem Biophys Res Commun279, 433 437]. In this report, we show that activation is effected by the product, Inosine monophosphate (IMP), and not by the substrates. Studies carried out on Plasmodium falciparum HGPRT and on a temperature-sensitive mutant, L44F, show that the enzymes are destabilized in the presence of the substrates and the product, IMP. These stability studies suggest that the active, product-bound form of the enzyme is less stable than the ligand-free, unactivated enzyme. Equilibrium isothermal unfolding studies indicate that the active form is destabilized by 2-3 kcal x mol(-1) compared with the unactivated state. This presents a unique example of an enzyme that attains its active conformation of lower stability by product binding. This property of ligand-mediated activation is not seen with recombinant human HGPRT, which is highly active in the unliganded state. The reversibility between highly active and weakly active states suggests a novel mechanism for the regulation of enzyme activity in P. falciparum. PMID- 15819885 TI - Differential effects of histone deacetylase inhibitors on phorbol ester- and TGF beta1 induced murine tissue inhibitor of metalloproteinases-1 gene expression. AB - Expression of the tissue inhibitor of metalloproteinases-1 (Timp-1) gene can be induced by either phorbol myristate acetate (PMA) or transforming growth factor beta1 (TGF-beta1), although the signalling pathways involved are not clearly defined. Canonically, histone deacetylase inhibitors (HDACi) such as trichostatin A (TSA) or sodium butyrate (NaB) increase total cellular histone acetylation and activate expression of susceptible genes. Remarkably, PMA and TGF-beta1 stimulation of Timp-1 show a differential response to TSA or NaB. TSA or NaB potentiate PMA-induced Timp-1 expression but repress TGF-beta1-induced Timp-1 expression. The repression of TGF-beta1-induced Timp-1 by TSA was maximal at 5 ng.mL(-1), while for the superinduction of PMA-induced Timp-1 expression, the maximal dose is > 500 ng x mL(-1) TSA. A further HDACi, valproic acid, did not block TGF-beta1-induced Timp-1 expression, demonstrating that different HDACs impact on the induction of Timp-1. For either PMA or TGF-beta1 to induce Timp-1 expression, new protein synthesis is required, and the induction of AP-1 factors closely precedes that of Timp-1. The effects of the HDACi can be reiterated in transient transfection using Timp-1 promoter constructs. Mutation or deletion of the AP-1 motif (-59/-53) in the Timp-1 promoter diminishes PMA-induction of reporter constructs, however, the further addition of TSA still superinduces the reporter. In c-Jun-/- cells, PMA still stimulates Timp-1 expression, but TSA superinduction is lost. Transfection of a series of Timp-1 promoter constructs identified three regions through which TSA superinduces PMA-induced Timp-1 and we have demonstrated specific protein binding to two of these regions which contain either an avian erythroblastosis virus E26 (v-ets) oncogene homologue (Ets) or Sp1 binding motif. PMID- 15819886 TI - Biochemical and spectroscopic characterization of the bacterial phytochrome of Pseudomonas aeruginosa. AB - Phytochromes are photochromic biliproteins found in plants as well as in some cyanotrophic, photoautotrophic and heterotrophic bacteria. In many bacteria, their function is largely unknown. Here we describe the biochemical and spectroscopic characterization of recombinant bacterial phytochrome from the opportunistic pathogen Pseudomonas aeruginosa (PaBphP). The recombinant protein displays all the characteristic features of a bonafide phytochrome. In contrast with cyanobacteria and plants, the chromophore of this bacterial phytochrome is biliverdin IXalpha, which is produced by the heme oxygenase BphO in P. aeruginosa. This chromophore was shown to be covalently attached via its A-ring endo-vinyl group to a cysteine residue outside the defined bilin lyase domain of plant and cyanobacterial phytochromes. Site-directed mutagenesis identified Cys12 and His247 as being important for chromophore binding and photoreversibility, respectively. PaBphP is synthesized in the dark in the red-light-absorbing Pr form and immediately converted into a far-red-light-absorbing Pfr-enriched form. It shows the characteristic red/far-red-light-induced photoreversibility of phytochromes. A chromophore analog that lacks the C15/16 double bond was used to show that this photoreversibility is due to a 15Z/15E isomerization of the biliverdin chromophore. Autophosphorylation of PaBphP was demonstrated, confirming its role as a sensor kinase of a bacterial two-component signaling system. PMID- 15819887 TI - Syndecan-4 is a signaling molecule for stromal cell-derived factor-1 (SDF-1)/ CXCL12. AB - Stromal cell-derived factor-1 (SDF-1)/CXCL12, the ligand for CXCR4, induces signal transduction. We previously showed that CXCL12 binds to high- and low affinity sites expressed by primary cells and cell lines, and forms complexes with CXCR4 as expected and also with a proteoglycan, syndecan-4, but does not form complexes with syndecan-1, syndecan-2, CD44 or beta-glycan. We also demonstrated the occurrence of a CXCL12-independent heteromeric complex between CXCR4 and syndecan-4. However, our data ruled out the glycosaminoglycan-dependent binding of CXCL12 to HeLa cells facilitating the binding of this chemokine to CXCR4. Here, we demonstrate that CXCL12 directly binds to syndecan-4 in a glycosaminoglycan-dependent manner. We show that upon stimulation of HeLa cells by CXCL12, CXCR4 becomes tyrosine phosphorylated as expected, while syndecan-4 (but not syndecan-1, syndecan-2 or beta-glycan) also undergoes such tyrosine phosphorylation. Moreover, tyrosine-phosphorylated syndecan-4 from CXCL12 stimulated HeLa cells physically coassociates with tyrosine phosphorylated CXCR4. Pretreatment of the cells with heparitinases I and III prevented the tyrosine phosphorylation of syndecan-4, which suggests that the heparan sulfate-dependent binding of SDF-1 to this proteoglycan is involved. Finally, by reducing syndecan 4 expression using RNA interference or by pretreating the cells with heparitinase I and III mixture, we suggest the involvement of syndecan-4 and heparan sulfate in p44/p42 mitogen-activated protein kinase and Jun N-terminal/stress-activated protein kinase activation by action of CXCL12 on HeLa cells. However, these treatments did not modify the calcium mobilization induced by CXCL12 in these cells. Therefore, syndecan-4 behaves as a CXCL12 receptor, selectively involved in some transduction pathways induced by SDF-1, and heparan sulfate plays a role in these events. PMID- 15819888 TI - Structures of Phanerochaete chrysosporium Cel7D in complex with product and inhibitors. AB - The cellobiohydrolase Pc_Cel7D is the major cellulase produced by the white-rot fungus Phanerochaete chrysosporium, constituting approximately 10% of the total secreted protein in liquid culture on cellulose. The enzyme is classified into family 7 of the glycoside hydrolases and, like other family members, catalyses cellulose hydrolysis with net retention of the anomeric carbon configuration. Previous work described the apo structure of the enzyme. Here we investigate the binding of the product, cellobiose, and several inhibitors, i.e. lactose, cellobioimidazole, Tris/HCl, calcium and a thio-linked substrate analogue, methyl 4-S-beta-cellobiosyl-4-thio-beta-cellobioside (GG-S-GG). The three disaccharides bind in the glucosyl-binding subsites +1 and +2, close to the exit of the cellulose-binding tunnel/cleft. Pc_Cel7D binds to lactose more strongly than cellobiose, while the opposite is true for the homologous Trichoderma reesei cellobiohydrolase Tr_Cel7A. Although both sugars bind Pc_Cel7D in a similar fashion, the different preferences can be explained by varying interactions with nearby loops. Cellobioimidazole is bound at a slightly different position, displaced approximately 2 A toward the catalytic centre. Thus the Pc_Cel7D complexes provide evidence for two binding modes of the reducing-end cellobiosyl moiety; this conclusion is confirmed by comparison with other available structures. The combined results suggest that hydrolysis of the glycosyl-enzyme intermediate may not require the prior release of the cellobiose product from the enzyme. Further, the structure obtained in the presence of both GG-S-GG and cellobiose revealed electron density for Tris at the catalytic centre. Inhibition experiments confirm that both Tris and calcium are effective inhibitors at the conditions used for crystallization. PMID- 15819889 TI - The multifarious short-term regulation of ammonium assimilation of Escherichia coli: dissection using an in silico replica. AB - Ammonium assimilation in Escherichia coli is regulated through multiple mechanisms (metabolic, signal transduction leading to covalent modification, transcription, and translation), which (in-)directly affect the activities of its two ammonium-assimilating enzymes, i.e. glutamine synthetase (GS) and glutamate dehydrogenase (GDH). Much is known about the kinetic properties of the components of the regulatory network that these enzymes are part of, but the ways in which, and the extents to which the network leads to subtle and quasi-intelligent regulation are unappreciated. To determine whether our present knowledge of the interactions between and the kinetic properties of the components of this network is complete - to the extent that when integrated in a kinetic model it suffices to calculate observed physiological behaviour - we now construct a kinetic model of this network, based on all of the kinetic data on the components that is available in the literature. We use this model to analyse regulation of ammonium assimilation at various carbon statuses for cells that have adapted to low and high ammonium concentrations. We show how a sudden increase in ammonium availability brings about a rapid redirection of the ammonium assimilation flux from GS/glutamate synthase (GOGAT) to GDH. The extent of redistribution depends on the nitrogen and carbon status of the cell. We develop a method to quantify the relative importance of the various regulators in the network. We find the importance is shared among regulators. We confirm that the adenylylation state of GS is the major regulator but that a total of 40% of the regulation is mediated by ADP (22%), glutamate (10%), glutamine (7%) and ATP (1%). The total steady state ammonium assimilation flux is remarkably robust against changes in the ammonium concentration, but the fluxes through GS and GDH are completely nonrobust. Gene expression of GOGAT above a threshold value makes expression of GS under ammonium-limited conditions, and of GDH under glucose-limited conditions, sufficient for ammonium assimilation. PMID- 15819890 TI - Introduction of extended LEC14-type branching into core-fucosylated biantennary N glycan. AB - A series of enzymatic substitutions modifies the basic structure of complex-type biantennary N-glycans. Among them, a beta1,2-linked N-acetylglucosamine residue is introduced to the central mannose moiety of the core-fucosylated oligosaccharide by N-acetylglucosaminyltransferase VII. This so-called LEC14 epitope can undergo galactosylation at the beta1,2-linked N-acetylglucosamine residue. Guided by the hypothesis that structural modifications in the N-glycan alter its capacity to serve as ligand for lectins, we prepared a neoglycoprotein with the extended LEC14 N-glycan and tested its properties in three different assays. In order to allow comparison to previous results on other types of biantennary N-glycans the functionalization of the glycans for coupling and assay conditions were deliberately kept constant. Compared to the core-fucosylated N glycan no significant change in affinity was seen when testing three galactoside specific proteins. However, cell positivity in flow cytofluorimetry was enhanced in six of eight human tumor lines. Analysis of biodistribution in tumor-bearing mice revealed an increase of blood clearance by about 40%, yielding a favorable tumor/blood ratio. Thus, the extended LEC14 motif affects binding properties to cellular lectins on cell surfaces and organs when compared to the core fucosylated biantennary N-glycan. The results argue in favor of the concept of viewing substitutions as molecular switches for lectin-binding affinity. Moreover, they have potential relevance for glycoengineering of reagents in tumor imaging. PMID- 15819891 TI - Small exterior hydrophobic cluster contributes to conformational stability and steroid binding in ketosteroid isomerase from Pseudomonas putida biotype B. AB - A structural motif called the small exterior hydrophobic cluster (SEHC) has been proposed to explain the stabilizing effect mediated by solvent-exposed hydrophobic residues; however, little is known about its biological roles. Unusually, in Delta(5)-3-ketosteroid isomerase from Pseudomonas putida biotype B (KSI-PI) Trp92 is exposed to solvent on the protein surface, forming a SEHC with the side-chains of Leu125 and Val127. In order to identify the role of the SEHC in KSI-PI, mutants of those amino acids associated with the SEHC were prepared. The W92A, L125A/V127A, and W92A/L125A/V127A mutations largely decreased the conformational stability, while the L125F/V127F mutation slightly increased the stability, indicating that hydrophobic packing by the SEHC is important in maintaining stability. The crystal structure of W92A revealed that the decreased stability caused by the removal of the bulky side-chain of Trp92 could be attributed to the destabilization of the surface hydrophobic layer consisting of a solvent-exposed beta-sheet. Consistent with the structural data, the binding affinities for three different steroids showed that the surface hydrophobic layer stabilized by SEHC is required for KSI-PI to efficiently recognize hydrophobic steroids. Unfolding kinetics based on analysis of the Phi(U) value also indicated that the SEHC in the native state was resistant to the unfolding process, despite its solvent-exposed site. Taken together, our results demonstrate that the SEHC plays a key role in the structural integrity that is needed for KSI-PI to stabilize the hydrophobic surface conformation and thereby contributes both to the overall conformational stability and to the binding of hydrophobic steroids in water solution. PMID- 15819892 TI - Poly(ADP-ribose) polymerase-1 protects excessive DNA strand breaks from deterioration during repair in human cell extracts. AB - Base excision repair (BER), a major pathway for the removal of simple lesions in DNA, requires the co-ordinated action of several repair and ancillary proteins, the impairment of which can lead to genetic instability. We here address the role of poly(ADP-ribose) polymerase-1 (PARP-1) in BER. Using an in vitro cross-linking assay, we reveal that PARP-1 is always involved in repair of a uracil-containing oligonucleotide and that it binds to the damaged DNA during the early stages of repair. Inhibition of PARP-1 poly(ADP-ribosyl)ation by 3-aminobenzamide blocks dissociation of PARP-1 from damaged DNA and prevents further repair. We find that excessive poly(ADP-ribosyl)ation occurs when repair intermediates containing single-strand breaks are in excess of the repair capacity of the cell extract, suggesting that repeated binding of PARP-1 to the nicked DNA occurs. We also find increased sensitivity of repair intermediates to nuclease cleavage in PARP deficient mouse fibroblasts and after depletion of PARP-1 from HeLa whole cell extracts. Our data support the model in which PARP-1 binding to DNA single-strand breaks or repair intermediates plays a protective role when repair is limited. PMID- 15819893 TI - Solution structure and internal dynamics of NSCP, a compact calcium-binding protein. AB - The solution structure of Nereis diversicolor sarcoplasmic calcium-binding protein (NSCP) in the calcium-bound form was determined by NMR spectroscopy, distance geometry and simulated annealing. Based on 1859 NOE restraints and 262 angular restraints, 17 structures were generated with a rmsd of 0.87 A from the mean structure. The solution structure, which is highly similar to the structure obtained by X-ray crystallography, includes two open EF-hand domains, which are in close contact through their hydrophobic surfaces. The internal dynamics of the protein backbone were determined by studying amide hydrogen/deuterium exchange rates and 15N nuclear relaxation. The two methods revealed a highly compact and rigid structure, with greatly restricted mobility at the two termini. For most of the amide protons, the free energy of exchange-compatible structural opening is similar to the free energy of structural stability, suggesting that isotope exchange of these protons takes place through global unfolding of the protein. Enhanced conformational flexibility was noted in the unoccupied Ca2+-binding site II, as well as the neighbouring helices. Analysis of the experimental nuclear relaxation and the molecular dynamics simulations give very similar profiles for the backbone generalized order parameter (S2), a parameter related to the amplitude of fast (picosecond to nanosecond) movements of N(H)-H vectors. We also noted a significant correlation between this parameter, the exchange rate, and the crystallographic B factor along the sequence. PMID- 15819894 TI - Actin mutations in hypertrophic and dilated cardiomyopathy cause inefficient protein folding and perturbed filament formation. AB - Hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM) are the most common hereditary cardiac conditions. Both are frequent causes of sudden death and are often associated with an adverse disease course. Alpha-cardiac actin is one of the disease genes where different missense mutations have been found to cause either HCM or DCM. We have tested the hypothesis that the protein-folding pathway plays a role in disease development for two actin variants associated with DCM and six associated with HCM. Based on a cell-free coupled translation assay the actin variants could be graded by their tendency to associate with the chaperonin TCP-1 ring complex/chaperonin containing TCP-1 (TRiC/CCT) as well as their propensity to acquire their native conformation. Some variant proteins are completely stalled in a complex with TRiC and fail to fold into mature globular actin and some appear to fold as efficiently as the wild-type protein. A fraction of the translated polypeptide became ubiquitinated and detergent insoluble. Variant actin proteins overexpressed in mammalian cell lines fail to incorporate into actin filaments in a manner correlating with the degree of misfolding observed in the cell-free assay; ranging from incorporation comparable to wild type actin to little or no incorporation. We propose that effects of mutations on folding and fiber assembly may play a role in the molecular disease mechanism. PMID- 15819895 TI - The structural comparison of the bacterial PepX and human DPP-IV reveals sites for the design of inhibitors of PepX activity. AB - X-prolyl dipeptidyl aminopeptidases (X-PDAP) are enzymes catalysing the release of dipeptides from the amino termini of polypeptides containing a proline or an alanine at the penultimate position. Involved in various mammalian regulation processes, as well as in chronic human diseases, they have been proposed to play a role in pathogenicity for Streptococci. We compared the structure of X-PDAP from Lactococcus lactis (PepX) with its human counterpart DPP-IV. Despite very different overall folds, the residues most implicated for X-PDAP activity are conserved in the same positions and orientations in both enzymes, thus defining a structural signature for the X-PDAP specificity that crosses the species frontiers of evolution. Starting from this observation, we tested some inhibitors of DPP-IV on PepX activity, for which no specific inhibitor is known. We thus found that PepX was highly sensitive to valine-pyrrolidide with a KI of 9.3 microm, close to that reported in DPP-IV inhibition. We finally used the structure of PepX from L. lactis as a template for computer-based homology modeling of PepX from the pathogenic Streptococcus gordonii. Docking simulations of valine-pyrrolidide into the active site of PepX led to the identification of key residues for a rational drug design against PepX from Streptococci. These results could have applications in human health giving new perspectives to the struggle against pathogens. PMID- 15819896 TI - Quaternary structure and functional properties of Penaeus monodon hemocyanin. AB - The hemocyanin of the tiger shrimp, Penaeus monodon, was investigated with respect to stability and oxygen binding. While hexamers occur as a major component, dodecamers and traces of higher aggregates are also found. Both the hexamers and dodecamers were found to be extremely stable against dissociation at high pH, independently of the presence of calcium ions, in contrast to the known crustacean hemocyanins. This could be caused by only a few additional noncovalent interactions between amino acids located at the subunit-subunit interfaces. Based on X-ray structures and sequence alignments of related hemocyanins, the particular amino acids are identified. At all pH values, the p50 and Bohr coefficients of the hexamers are twice as high as those of dodecamers. While the oxygen binding of hexamers from crustaceans can normally be described by a simple two-state model, an additional conformational state is needed to describe the oxygen-binding behaviour of Penaeus monodon hemocyanin within the pH range of 7.0 to 8.5. The dodecamers bind oxygen according to the nested Monod-Whyman-Changeaux (MWC) model, as observed for the same aggregation states of other hemocyanins. The oxygen-binding properties of both the hexameric and dodecameric hemocyanins guarantee an efficient supply of the animal with oxygen, with respect to the ratio between their concentrations. It seems that under normoxic conditions, hexamers play the major role. Under hypoxic conditions, the hexamers are expected not to be completely loaded with oxygen. Here, the dodecamers are supposed to be responsible for the oxygen supply. PMID- 15819897 TI - Hyperthermal stability of neuroglobin and cytoglobin. AB - Neuroglobin (Ngb) and cytoglobin (Cygb), recent additions to the globin family, display a hexa-coordinated (bis-histidyl) heme in the absence of external ligands. Although these proteins have the classical globin fold they reveal a very high thermal stability with a melting temperature (Tm) of 100 degrees C for Ngb and 95 degrees C for Cygb. Moreover, flash photolysis experiments at high temperatures reveal that Ngb remains functional at 90 degrees C. Human Ngb may have a disulfide bond in the CD loop region; reduction of the disulfide bond increases the affinity of the iron atom for the distal (E7) histidine, and leads to a 3 degrees C increase in the T(m) for ferrous Ngb. A similar Tm is found for a mutant of human Ngb without cysteines. Apparently, the disulfide bond is not involved directly in protein stability, but may influence the stability indirectly because it modifies the affinity of the distal histidine. Mutation of the distal histidine leads to lower thermal stability, similar to that for other globins. Only globins with a high affinity of the distal histidine show the very high thermal stability, indicating that stable hexa-coordination is necessary for the enhanced thermal stability; the CD loop which contains the cysteines appears as a critical region in the neuroglobin thermal stability, because it may influence the affinity of the distal histidine. PMID- 15819898 TI - Leaf form and the reconstruction of past climates. PMID- 15819899 TI - Unearthing ectomycorrhizal dynamics. PMID- 15819900 TI - Linking mutation to adaptation: overcoming stress at the spa. PMID- 15819901 TI - Linking tocopherols with cellular signaling in plants. PMID- 15819902 TI - Data-model integration is not magic. PMID- 15819903 TI - Sulfur and phytoplankton: acquisition, metabolism and impact on the environment. AB - Sulfur emission from marine phytoplankton has been recognized as an important factor for global climate and as an entry into the biogeochemical S cycle. Despite this significance, little is known about the cellular S metabolism in algae that forms the basis of this emission. Some biochemical and genetic evidence for regulation of S uptake and assimilation is available for the freshwater model alga Chlamydomonas. However, the marine environment is substantially different from most fresh waters, containing up to 50 times higher free sulfate concentrations and challenging the adaptive mechanisms of primary and secondary S metabolism in marine algae. This review intends to integrate ecological and physiological data to provide a comprehensive view of the role of S in the oceans. PMID- 15819904 TI - Form, function and environments of the early angiosperms: merging extant phylogeny and ecophysiology with fossils. AB - The flowering plants--angiosperms--appeared during the Early Cretaceous period and within 10-30 Myr dominated the species composition of many floras worldwide. Emerging insights into the phylogenetics of development and discoveries of early angiosperm fossils are shedding increased light on the patterns and processes of early angiosperm evolution. However, we also need to integrate ecology, in particular how early angiosperms established a roothold in pre-existing Mesozoic plant communities. These events were critical in guiding subsequent waves of angiosperm diversification during the Aptian-Albian. Previous pictures of the early flowering plant ecology have been diverse, ranging from large tropical rainforest trees, weedy drought-adapted and colonizing shrubs, disturbance- and sun-loving rhizomatous herbs, and, more recently, aquatic herbs; however, none of these images were tethered to a robust hypothesis of angiosperm phylogeny. Here, we synthesize our current understanding of early angiosperm ecology, focusing on patterns of functional ecology, by merging recent molecular phylogenetic studies and functional studies on extant 'basal angiosperms' with the picture of early angiosperm evolution drawn by the fossil record. PMID- 15819905 TI - Ancient plant DNA: review and prospects. AB - Ancient DNA has received much attention since the mid-1980s, when the first sequence of an extinct animal species was recovered from a museum specimen. Since then, the majority of ancient DNA studies have focused predominantly on animal species, while studies in plant palaeogenetics have been rather limited, with the notable exception of cultivated species found in archaeological sites. Here, we outline the recent developments in the analysis of plant ancient DNA. We emphasize the trend from species identification to population-level investigation and highlight the potential and the difficulties in this field, related to DNA preservation and to risks of contamination. Further efforts towards the analysis of ancient DNA from the abundant store of fossil plant remains should provide new research opportunities in palaeoecology and phylogeography. In particular, intraspecific variation should be considered not only in cultivated plants but also in wild taxa if palaeogenetics is to become a fully emancipated field of plant research. PMID- 15819906 TI - Isoprene decreases the concentration of nitric oxide in leaves exposed to elevated ozone. AB - Isoprene reduces visible damage (necrosis) of leaves caused by exposure to ozone but the mechanism is not known. Here we show that in Phragmites leaves isoprene emission was stimulated after a 3-h exposure to high ozone levels. The photosynthetic apparatus of leaves in which isoprene emission was inhibited by fosmidomycin became more susceptible to damage by ozone than in isoprene-emitting leaves. Three days after ozone fumigation, the necrotic leaf area was significantly higher in isoprene-inhibited leaves than in isoprene-emitting leaves. Isoprene-inhibited leaves also accumulated high amounts of nitric oxide (NO), as detected by epifluorescence light microscopy. Our results confirm that oxidative stresses activate biosynthesis and emission of chloroplastic isoprenoid, bringing further evidence in support of an antioxidant role for these compounds. It is suggested that, in nature, the simultaneous quenching of NO and reactive oxygen species by isoprene may be a very effective mechanism to control dangerous compounds formed under abiotic stress conditions, while simultaneously attenuating the induction of the hypersensitive response leading to cellular damage and death. PMID- 15819907 TI - The light-hyperresponsive high pigment-2dg mutation of tomato: alterations in the fruit metabolome. AB - Overall metabolic modifications between fruit of light-hyperresponsive high pigment (hp) tomato (Lycopersicon esculentum) mutant plants and isogenic nonmutant (wt) control plants were compared. Targeted metabolite analyses, as well as large-scale nontargeted mass spectrometry (MS)-based metabolite profiling, were used to phenotype the differences in fruit metabolite composition. Targeted high-performance liquid chromatography with photodiode array detection (HPLC-PDA) metabolite analyses showed higher levels of isoprenoids and phenolic compounds in hp-2dg fruit. Nontargeted GC-MS profiling of red fruits produced 25 volatile compounds that showed a 1.5-fold difference between the genotypes. Analyses of red fruits using HPLC coupled to high resolution quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) in both ESI positive and ESI-negative mode generated, respectively, 6168 and 5401 mass signals, of which 142 and 303 showed a twofold difference between the genotypes. hp-2dg fruits are characterized by overproduction of many metabolites, several of which are known for their antioxidant or photoprotective activities. These metabolites may now be more closely implicated as resources recruited by plants to respond to and manage light stress. The similarity in metabolic alterations in fruits of hp-1 and hp-2 mutant plants helps us to understand how hp mutations affect cellular processes. PMID- 15819908 TI - The rhizosphere signal molecule lumichrome alters seedling development in both legumes and cereals. AB - The stimulatory role of lumichrome, a rhizosphere metabolite, was assessed on the growth of legume and cereal seedlings. At a very low nanomolar concentration (5 nm), lumichrome elicited growth promotion in cowpea, soybean, sorghum, millet and maize, but not in common bean, Bambara groundnut and Sudan grass. In soybean and cowpea only, 5 nm lumichrome caused early initiation of trifoliate leaf development, expansion in unifoliate and trifoliate leaves, increased stem elongation and, as a result, an increase in shoot and plant total biomass relative to control. Lumichrome (5 nm) also increased leaf area in maize and sorghum, and thus raised shoot and total biomass but there was no effect on the leaf area of the other cereals. Root growth was also stimulated in sorghum and millet by the supply of 5 nm lumichrome. By contrast, the application of a higher dose of lumichrome (50 nm) depressed development of unifoliate leaves in soybean, the second trifoliate leaf in cowpea, and shoot biomass in soybean. The 50 nm concentration also consistently decreased root development in cowpea and millet, but had no effect on the other species. These data show that lumichrome is a rhizosphere signal molecule that affects seedling development in both monocots and dicots. PMID- 15819909 TI - Induced plant uptake and transport of mercury in the presence of sulphur containing ligands and humic acid. AB - The induced accumulation of mercury (Hg) by plants was investigated for the species Phaseolus vulgaris (Bush bean), Brassica juncea (Indian mustard), and Vicia villosa (Hairy vetch). All plants were grown in modified Hg-contaminated mine tailings and were treated with sulphur-containing ligands to induce Hg accumulation. The effects of varied substrate Hg concentration and humic acid (HA) level on the induced plant-Hg accumulation for B. juncea were examined. Thiosulphate salts (ammonium and sodium) mobilised Hg in the substrates and caused an increase in the Hg concentration of roots and shoots of all tested plant species. Root Hg accumulation was positively correlated to extractable Hg for (NH4)2S2O3-treated B. juncea plants grown in HA-amended substrates. However, shoot Hg translocation for this species was inhibited at 1.25 g HA kg(-1) of substrate. Mercury-thiosulphate complexes could be translocated and accumulated in the upper parts of the plants up to 25 times the Hg concentration in the substrate. We conclude that shoot Hg accumulation in the presence of thiosulphate salts is dependent upon plant species characteristics (e.g. root surface area) and humic acid content. PMID- 15819910 TI - Genetic characterization of androgenic progeny derived from Lolium perenne x Festuca pratensis cultivars. AB - Androgenesis and genetic characterization of androgenic progeny from Lolium perenne x Festuca pratensis cultivars (2n = 4x = 28) were investigated in order to develop a novel grass that combines favourable attributes of parent plants. A successful androgenesis was obtained using PG-96 medium. The green plant regeneration was 46%, 35% and 17% for Bx350, Bx351 and Prior, respectively, and over 800 green plants have been obtained. Androgenic progeny showed a large variation in freezing tolerance, 7% of 292 progeny exceeding that of freezing hardy F. pratensis despite containing chromosomes of L. perenne. More than 60% of 175 flowering progeny produced dehiscent anthers with pollens. Androgenic plants contained 14 or 28 chromosomes. There were 188 (56%), 204 (77%) and 114 (81%) dihaploids from Bx350, Bx351 and Prior, respectively. The nuclear DNA content varied significantly even between plants with the same chromosome number. High levels of chromosome pairing and recombination were observed by genomic in situ hybridization analysis because of close homology between two genomes. The results indicated that androgenic progeny of Festulolium showed high level of genetic variation, and provide potential for accelerating selection efficiency. PMID- 15819911 TI - Environmental signals from leaves--a physiognomic analysis of European vegetation. AB - Leaf physiognomic traits vary predictably along climatic and environmental gradients. The relationships between leaf physiognomy and climate have been investigated on different continents, but so far an investigation based on European vegetation has been missing. A grid data set (0.5 degrees x 0.5 degrees latitude/longitude) has been compiled in order to determine spatial patterns of leaf physiognomy across Europe. Based on distribution maps of native European hardwoods, synthetic chorologic flora lists were compiled for all grid cells. Every synthetic chorologic flora was characterised by 25 leaf physiognomic traits and correlated with 16 climatic parameters. Clear spatial patterns of leaf physiognomy have been observed, which are statistically significant related to certain, temperature-related climate parameters. Transfer functions for several climatic parameters have been established, based on the observed relationships. The study provides evidence that synthetically generated floras represent a powerful tool for analysing spatial patterns of leaf physiognomy and their relationships to climate. The transfer functions from the European data set indicate slightly different relationships of leaf physiognomy and environment compared with results obtained from other continents. PMID- 15819913 TI - Variation in the degree of coupling between delta13C of phloem sap and ecosystem respiration in two mature Nothofagus forests. AB - Day-to-day variability in the carbon isotope composition of phloem sap (delta13Chd) and ecosystem respiratory CO2 (delta13CR) were measured to assess the tightness of coupling between canopy photosynthesis (delta13Chd) and ecosystem respiration (delta13CR) in two mature Nothofagus solandri (Hook. f.) forests in New Zealand. Abundant phloem-tapping scale insects allowed repeated, nondestructive access to stem phloem sap 1-2 m above ground. delta13Chd was compared with delta13C predicted by an environmentally driven, process-based canopy photosynthesis model. Keeling plots of within-canopy CO2 were used to estimate delta13CR. By including a lag of 3 d, there was good agreement in the timing and direction of variation in delta13Chd and predictions by the canopy photosynthesis model, suggesting that delta13Chd represents a photosynthesis weighted, integrative record of canopy photosynthesis and conductance. Significant day-to-day variability in delta13CR was recorded at one of the two forests. At this site, delta13CR reflected variability in delta13Chd only on days with <2 mm rain. We conclude that the degree of coupling between canopy photosynthesis and ecosystem respiration varies between sites, and with environmental conditions at a single site. PMID- 15819912 TI - Assessing the generality of global leaf trait relationships. AB - Global-scale quantification of relationships between plant traits gives insight into the evolution of the world's vegetation, and is crucial for parameterizing vegetation-climate models. A database was compiled, comprising data for hundreds to thousands of species for the core 'leaf economics' traits leaf lifespan, leaf mass per area, photosynthetic capacity, dark respiration, and leaf nitrogen and phosphorus concentrations, as well as leaf potassium, photosynthetic N-use efficiency (PNUE), and leaf N : P ratio. While mean trait values differed between plant functional types, the range found within groups was often larger than differences among them. Future vegetation-climate models could incorporate this knowledge. The core leaf traits were intercorrelated, both globally and within plant functional types, forming a 'leaf economics spectrum'. While these relationships are very general, they are not universal, as significant heterogeneity exists between relationships fitted to individual sites. Much, but not all, heterogeneity can be explained by variation in sample size alone. PNUE can also be considered as part of this trait spectrum, whereas leaf K and N : P ratios are only loosely related. PMID- 15819914 TI - Atmospheric CO2 enrichment alters energy assimilation, investment and allocation in Xanthium strumarium. AB - Energy-use efficiency and energy assimilation, investment and allocation patterns are likely to influence plant growth responses to increasing atmospheric CO2 concentration ([CO2]). Here, we describe the influence of elevated [CO2] on energetic properties as a mechanism of growth responses in Xanthium strumarium. Individuals of X. strumarium were grown at ambient or elevated [CO2] and harvested. Total biomass and energetic construction costs (CC) of leaves, stems, roots and fruits and percentage of total biomass and energy allocated to these components were determined. Photosynthetic energy-use efficiency (PEUE) was calculated as the ratio of total energy gained via photosynthetic activity (Atotal) to leaf CC. Elevated [CO2] increased leaf Atotal, but decreased CC per unit mass of leaves and roots. Consequently, X. strumarium individuals produced more leaf and root biomass at elevated [CO2] without increasing total energy investment in these structures (CCtotal). Whole-plant biomass was associated positively with PEUE. Whole-plant construction required 16.1% less energy than modeled whole-plant energy investment had CC not responded to increased [CO2]. As a physiological mechanism affecting growth, altered energetic properties could positively influence productivity of X. strumarium, and potentially other species, at elevated [CO2]. PMID- 15819915 TI - Differential effects of interspecific interactions and water availability on survival, growth and fecundity of three congeneric grassland herbs. AB - Fitness of individual plants and of populations depends on the rates of survival, growth and fecundity. This study tested whether vital rates were differentially affected by biotic interactions and water availability. The effects of manipulations of above-ground competition (through clipping) and water availability (through water addition) on the vital rates of seedlings of three species (Viola elatior Fries, Viola pumila Chaix and Viola stagnina Kit.) were analysed in dry, mesic and wet grasslands. Water addition and grassland type had the largest effects on survival (accounting for 41 and 24% of total variation, respectively) across species. Height growth rate was positively affected by grassland type (19%) and water addition (12%) and varied among species (8%), while leaf accumulation rates and reproduction were affected by grassland type and clipping. The data suggested facilitative effects of the canopy on seedling survival in the dry grassland. This study presents evidence that environmental conditions and biotic interactions may have differential effects on seedling survival, growth and reproduction. The findings highlight the complex interplay between spatial and temporal environmental variation and biotic interactions in structuring plant communities. PMID- 15819916 TI - High nitrogen : phosphorus ratios reduce nutrient retention and second-year growth of wetland sedges. AB - Shifts from nitrogen (N)- to phosphorus (P)-limited growth due to high N deposition may alter the functioning of wetland vegetation. This experiment tested how N vs P deficiency affects the growth and nutrient use of wetland sedges. Five wetland Carex species were grown at nine N : P supply ratios (0.6 405) with two absolute levels of N and P. Biomass and nutrient concentrations were determined after one and two growing seasons. Shoot biomass was maximal at N : P supply ratios of 15-26 after one season but 5-15 after two seasons. Photosynthesis after the first season, second-year growth, leaf longevity, and the fraction of nutrient supply retained by plants over two seasons were all negatively related to N : P supply ratios, with small effects of absolute supply. The five Carex species responded similarly to N : P ratios but differed in nutrient resorption efficiency and biomass allocation. Plants treated with high N : P ratios appeared to lose nutrients below ground. Such losses may reduce plant performance in P-limited wetlands affected by high N deposition. PMID- 15819917 TI - Decline of acid-sensitive plant species in heathland can be attributed to ammonium toxicity in combination with low pH. AB - The effects of increasing ammonium concentrations in combination with different pH levels were studied on five heathland plant species to determine whether their occurrence and decline could be attributed to ammonium toxicity and/or pH levels. Plants were grown in growth media amended with four different ammonium concentrations (10, 100, 500 and 1000 micromol l(-1)) and two pH levels resembling acidified (pH 3.5 or 4) and weakly buffered (pH 5 or 5.5) situations. Survival of Antennaria dioica and Succisa pratensis was reduced by low pH in combination with high ammonium concentrations. Biomass decreased with increased ammonium concentrations and decreasing pH levels. Internal pH of the plants decreased with increasing ammonium concentrations. Survival of Calluna vulgaris, Deschampsia flexuosa and Gentiana pneumonanthe was not affected by ammonium. Moreover, biomass increased with increasing ammonium concentrations. Biomass production of G. pneumonanthe reduced at low pH levels. A decline of acid sensitive species in heathlands was attributed to ammonium toxicity effects in combination with a low pH. PMID- 15819918 TI - The importance of nitrogen and carbohydrate storage for plant growth of the alpine herb Veratrum album. AB - We examined whether nitrogen (N) and carbohydrates reserves allow Veratrum album, an alpine forb, to start spring growth earlier than the neighbouring vegetation and to survive unpredictable disturbances resulting in loss of above-ground biomass. * Seasonal dynamics of plant reserves, soil N availability and vegetation growth were monitored. Veratrum album shoots were experimentally removed when carbohydrate reserves were at a seasonal minimum and the subsequent changes in biomass and reserves were compared with those in control plants. Reserves did not give V. album a competitive advantage in spring; however, they did function as a buffer against the impact of calamities. Shoot removal resulted in significantly lower root dry weight, higher N concentration in rhizome and roots and lower starch concentrations in rhizome and roots but no plant mortality was observed. Veratrum album used stored N reserves to supplement N uptake and establish high leaf N concentrations, which facilitated a rapid refilling of depleted carbohydrate reserves. The primary function of N reserves appears to be to allow V. album to complete the growing cycle in as short a period as possible, thus minimizing exposure to above-ground risks. PMID- 15819919 TI - Phyllosphere nitrogen relations: reciprocal transfer of nitrogen between epiphyllous liverworts and host plants in the understorey of a lowland tropical wet forest in Costa Rica. AB - Epiphyllous bryophytes on tropical rainforest plants acquire nutrients from throughfall and free-living N2-fixing organisms, but may also depend directly on host leaf leachates. By contrast, after drying events bryophytes lose significant quantities of nutrients through leaching that can be taken up by host leaves. To assess a potential nutritional interdependency, nitrogen fluxes between epiphyllous liverworts and their host leaves (Carludovica drudei, Costus laevis, Dieffenbachia concinna, Pentagonia wendlandii) were quantified by in situ15N labelling techniques in a lowland rainforest, Piedras Blancas National Park, Costa Rica. Depending on host species, epiphyllous bryophytes met between 1 and 57% of their N demand from host leaf leachates. Externally supplied 15N was taken up both by epiphylls and host leaves, but N from epiphyll leachates accounted for < 2.5% of host leaf N after 14 d. Long-term observations (180 d) demonstrated the highly dynamic nature of phyllosphere N of the investigated tropical rainforest understorey and an intermittent sink capacity of epiphyllous bryophytes. PMID- 15819920 TI - Plant-microbe interactions: identification of epiphytic bacteria and their ability to alter leaf surface permeability. AB - Bacteria were either isolated from leaf surfaces of Hedera helix or obtained from a culture collection in order to analyse their effect on barrier properties of isolated Hedera and Prunus laurocerasus cuticles. On the basis of the 16S rDNA sequences the genera of the six bacterial isolates from Hedera were identified as Pseudomonas sp., Stenotrophomonas sp. and Achromobacter. Water permeability of cuticles isolated from H. helix was measured before and after inoculation with the six bacterial strains. In addition water permeability of cuticles isolated from P. laurocerasus was measured before and after inoculation with the three bacterial strains Pseudomonas aeruginosa, Xanthomonas campestris and Corynebacterium fascians. Rates of water diffusing across isolated cuticles of both species significantly increased by up to 50% after inoculation with all bacterial strains. Obtained results show that epiphytic bacteria have the ability of increasing water permeability of Hedera and Prunus cuticles, which in turn should increase the availability of water and dissolved compounds in the phyllopshere. Consequently, living conditions in the habitat phyllosphere are improved. It can be concluded that the ability to change leaf surface properties will improve epiphytic fitness of leaf surface bacteria. PMID- 15819921 TI - Radiocarbon--a low-impact tool to study nutrient transport by soil fungi under field conditions. AB - Here, we present a new in-situ method to study the uptake of amino acids by soil fungi. We injected 14C-labeled glycine into a marshland soil and measured the rate and the 14C signature of CO2 respired from sporocarps of Pholiota terrestris over 53.5 h and 2 m. We also determined the incorporation of glycine-C into sporocarp tissue. The 14C signature of the CO2 and tissue was quantified by accelerator mass spectrometry. After the label application, the rate of CO2 flux and its 14C signature from chambers with sporocarps were significantly higher than from chambers without sporocarps, and then declined with time. Postlabel, the 14C signature of the sporocarp tissue increased by 35 per thousand. We show that this approach can be used to study below-ground food webs on an hourly time scale while minimizing the perturbation of competitive relationships among soil microorganisms and between plants and soil microorganisms. Additionally we show that care must be taken to avoid confounding effects of sporocarp senescence on rates and radiocarbon signatures of respired CO2. PMID- 15819922 TI - Does the presence of arbuscular mycorrhizal fungi influence growth and nutrient uptake of a wild-type tomato cultivar and a mycorrhiza-defective mutant, cultivated with roots sharing the same soil volume? AB - We investigated the growth and nutrient uptake of the Lycopersicon esculentum symbiosis mycorrhiza-defective plant mutant rmc, challenged with arbuscular mycorrhiza (AM) fungal propagules, in the presence or absence of roots of the commercial wild-type tomato cv. Golden Queen (GQ). Two plants shared the middle (combi) compartment of a horizontal three-compartment split-root pot with one part of their root system; the other part was grown separately in an outer (solo) pot. Combinations of rmc and GQ plants were grown together in soil that was either mycorrhiza-free (-M) or prepared with AM fungal inoculum (+M). Surface colonization of rmc roots was strongly increased in the presence of (+M) GQ roots. AM fungal inoculation increased phosphorus uptake of GQ plants, but decreased growth and P uptake of rmc plants. Growth and P uptake of (+M) GQ plants were reduced when plants were grown in combination with rmc rather than another GQ plant. AM fungi in the (combi) compartment may have preferentially formed hyphae spreading infection rather than functioning in P uptake in (+M) GQ plants grown in combination with rmc. Surface colonization of (+M) rmc roots, in the presence of GQ roots, was probably established at the expense of carbohydrates from associated GQ plants. Possible reasons for a decreased P uptake of rmc plants in response to AM fungal inoculation are proposed. PMID- 15819923 TI - Metabolic activity of Glomus intraradices in Arum- and Paris-type arbuscular mycorrhizal colonization. AB - Colonization of two plant species by Glomus intraradices was studied to investigate the two morphological types (Arum and Paris), their symbiotic interfaces and metabolic activities. Root pieces and sections were stained to observe the colonization and metabolic activity of all mycorrhizal structures. There were no growth responses observed in the plants caused by mycorrhizal symbiosis. The two morphological types had a similar percentage of root colonized, but the Arum-type had higher metabolic activity. Most of the mycorrhizal structures (88%) showed succinate dehydrogenase activity; about half showed acid phosphatase activity; and a small percentage showed alkaline phosphatase activity. Phosphatase activity was highest in arbuscules and low in intercellular hyphae in the Arum-type colonization. In the Paris-type, hyphal coils and arbusculate coils showed a similar intermediate percentage of phosphatase activity. We conclude that acid phosphatase is more important than alkaline phosphatase in both colonization types. We discuss the possibility that, whereas arbuscules in Arum-type are the main site for phosphorus release to the host plant, both the hyphal and arbusculate coils may be involved in the Paris type. PMID- 15819924 TI - Detection of plot-level changes in ectomycorrhizal communities across years in an old-growth mixed-conifer forest. AB - Understanding spatial and temporal patterns present in ectomycorrhizal fungal community structure is critical to understanding both the scale and duration of the potential impact these fungi have on the plant community. While recent studies consider the spatial structure of ectomycorrhizal communities, few studies consider how this changes over time. Ectomycorrhizal root biomass and the similarity of community composition were measured at scales up to 20 cm replicated in nine plots and over 3 yr. Soil cores were additionally stratified into three depths. Annual occurrence of the dominant ectomycorrhizal species was constant at larger spatial scales but varied more across years at a fine spatial scale. Turnover of ectomycorrhizal species between years was observed frequently at scales < 20 cm. The ectomycorrhizal community within a plot was more similar across years than it was to other plots sampled in the same year. Our results demonstrate the dynamic nature of the ectomycorrhizal community even in the absence of large-scale disturbances. The potential role of root turnover and drought stress is discussed. PMID- 15819925 TI - Priority effects determine the outcome of ectomycorrhizal competition between two Rhizopogon species colonizing Pinus muricata seedlings. AB - Competition is often considered a fundamental process influencing assemblage structure, yet little is known about competition among ectomycorrhizal (EM) fungi. Here, we examine competitive interactions between Rhizopogon occidentalis and Rhizopogon salebrosus in a 6-month microcosm study. Pinus muricata seedlings were grown in three EM treatments: R. occidentalis, R. salebrosus, and R. occidentalis and R. salebrosus. At 2, 4, and 6 months, five seedlings per treatment were harvested and the EM root tip biomass of each species was determined. Root tips in the two-species treatment were identified using molecular techniques. R. occidentalis had similar EM root tip biomass when grown alone or in the presence of R. salebrosus. By contrast, R. salebrosus had significantly lower EM root tip biomass when grown with R. occidentalis than when grown alone, indicating it was a competitive inferior under the conditions tested. Competition was driven by differences in timing of colonization resulting in a strong priority effect for R. occidentalis. Our results, together with two earlier studies, indicate competition may play a more important role in EM interactions than previously recognized. PMID- 15819926 TI - Mixotrophy in orchids: insights from a comparative study of green individuals and nonphotosynthetic individuals of Cephalanthera damasonium. AB - Some green orchids obtain carbon (C) from their mycorrhizal fungi and photosynthesis. This mixotrophy may represent an evolutionary step towards mycoheterotrophic plants fully feeding on fungal C. Here, we report on nonphotosynthetic individuals (albinos) of the green Cephalanthera damasonium that likely represent another evolutionary step. Albino and green individuals from a French population were compared for morphology and fertility, photosynthetic abilities, fungal partners (using microscopy and molecular tools), and nutrient sources (as characterized by 15N and 13C abundances). Albinos did not differ significantly from green individuals in morphology and fertility, but tended to be smaller. They harboured similar fungi, with Thelephoraceae and Cortinariaceae as mycorrhizal partners and few rhizoctonias. Albinos were nonphotosynthetic, fully mycoheterotrophic. Green individuals carried out photosynthesis at compensation point and received almost 50% of their C from fungi. Orchid fungi also colonized surrounding tree roots, likely to be the ultimate C source. Transition to mycoheterotrophy may require several simultaneous adaptations; albinos, by lacking some of them, may have reduced ecological success. This may limit the appearance of cheaters in mycorrhizal networks. PMID- 15819927 TI - Adaptation of Spirogyra insignis (Chlorophyta) to an extreme natural environment (sulphureous waters) through preselective mutations. AB - Adaptation of Spirogyra insignis (Chlorophyceae) to growth and survival in an extreme natural environment (sulphureous waters from La Hedionda Spa, S. Spain) was analysed by using an experimental model. Photosynthesis and growth of the alga were inhibited when it was cultured in La Hedionda Spa waters (LHW), but after further incubation for several weeks, the culture survived due to the growth of a variant that was resistant to LHW. A Luria-Delbruck fluctuation analysis was carried out to distinguish between resistant filaments arising from rare spontaneous mutations and resistant filaments arising from other mechanisms of adaptation. It was demonstrated that the resistant filaments arose randomly by rare spontaneous mutations before the addition of LHW (preselective mutations). The rate of spontaneous mutation from sensitivity to resistance was 2.7 x 10(-7) mutants per cell division. Since LHW(resistant) mutants have a diminished growth rate, they are maintained in nonsulphureous natural waters as the result of a balance between new resistants arising from spontaneous mutation and resistants eliminated by natural selection. Thus, recurrence of rare spontaneous preselective mutations ensures the survival of the alga in sulphureous waters. PMID- 15819928 TI - Local genetic differentiation among populations of the mass-flowering tropical shrub Erythroxylum havanense (Erythroxylaceae). AB - This study is aimed to understand the role of life-history traits in determining the genetic structure of populations. We used Erythroxylum havanense, a distylous shrub with synchronous and massive patterns of flowering and fruiting. We suggest that the high concentration of ephemeral resources produced by mass flowering satiates both pollinators and frugivores, restricting gene flow and leading to genetic differentiation among populations. Using random amplified polymorphic DNAs as genetic markers, we estimated genetic diversity and structure statistics to quantify the amount and distribution of genetic variation within and among five populations from the Pacific coast of Mexico. High levels of genetic variation within populations and significant differentiation among populations located very near to each other were found. Furthermore, spatial autocorrelation analysis indicated the presence of significant genetic structure at short spatial distances. We suggest that by influencing the foraging behavior of pollinators and frugivores, mass flowering may produce the observed patterns of genetic structure, while small differences in flowering or fruiting phenology could further reinforce the isolation of nearby populations. PMID- 15819929 TI - Reproductive compensation in the evolution of plant mating systems. AB - Reproductive compensation, the replacement of dead embryos by potentially viable ones, is known to play a major role in the maintenance of deleterious mutations in mammalian populations. However, it has received little attention in plant evolution. Here we model the joint evolution of mating system and inbreeding depression with reproductive compensation. We used a dynamic model of inbreeding depression, allowing for partial purging of recessive lethal mutations by selfing. We showed that reproductive compensation tended to increase the mean number of lethals in a population, but favored self-fertilization by effectively decreasing early inbreeding depression. When compensation depended on the selfing rate, stable mixed mating systems can occur, with low to intermediate selfing rates. Experimental evidence of reproductive compensation is required to confirm its potential importance in the evolution of plant mating systems. We suggest experimental methods to detect reproductive compensation. PMID- 15819930 TI - Maternal effects and heterosis influence the fitness of plant hybrids. AB - Here we tested two possible nonexclusive explanations for the maintenance of a hybrid swarm between Senecio jacobaea and Senecio aquaticus; first, that genotype by-environment interactions involving water and nutrient clines are involved in hybrid fitness, and second, heterosis in early hybrid generations may provide an initial hybrid advantage that contributes to hybrid persistence. In three climate chamber studies, fitness and root growth were measured for parental species and natural and artificial F1 hybrids, in order to determine whether hybrids occur in habitats where they are more fit than parental species. Natural hybrids, which are generally back-crossed to S. jacobaea, always equaled S. jacobaea in growth characteristics. Maternal effects played a role in the fitness of F1 hybrids, with offspring from S. jacobaea mothers exhibiting higher fitness than those from S. aquaticus mothers, and compared with parental species and natural hybrids. Natural hybrids are not distributed in zones where they are most fit with respect to nutrient and water regimes. Superior fitness of early generation hybrids may contribute to hybrid swarm stability. PMID- 15819931 TI - Laser scanning reveals bryophyte canopy structure. AB - We evaluated laser scanning as a method to provide depth measurements for bryophyte canopies at fine spatial scales to derive surface roughness (Lr), a structural parameter. Depths to the first vertical canopy contact were measured on 5 x 5 cm2 areas of 27 bryophyte canopies using a contact probe, a commercial laser scanner and a scanner employing a laser diode striper (LED scanner). Laser scanning adequately distinguished structural types, but scanner configuration led to differences in the magnitude of Lr. LED scanning did not damage photosystem II function in three bryophyte species, Bazzania trilobata, Sphagnum girgensohnii and Pleurozium schreberi, as evidenced by no change in the chlorophyll fluorescence parameter FV/FM following LED scanning, but a decrease when subjected to high light. A previously published boundary layer conductance model was parameterized with surface roughness values determined using a laser scanner and compared with the results obtained with contact probe measures. The resulting parameters of the functional models did not differ significantly from each other. PMID- 15819932 TI - The silent revolution of intensive ground water use: pros and cons. PMID- 15819937 TI - Ground Water Vistas: a graphical user interface for the MODFLOW family of ground water flow and transport models. PMID- 15819938 TI - Plume and lithologic profiling with surface resistivity and seismic tomography. AB - Improved surface-based geophysical technologies that are commercially available provide a new level of detail that can be used to guide ground water remediation. Surface-based multielectrode resistivity methods and tomographic seismic refraction techniques were used to image to a depth of approximately 30 m below the surface at the Natural and Accelerated Bioremediation Research Field Research Center. The U.S. Department of Energy (DOE) established the research center on the DOE Oak Ridge Reservation in Oak Ridge, Tennessee, to conduct in situ field scale studies on bioremediation of metals and radionuclides. Bioremediation studies are being conducted on the saprolite, shale bedrock, and ground water at the site that have been contaminated with nitrate, uranium, technetium, tetrachloroethylene, and other contaminants (U.S. DOE 1997). Geophysical methods were effective in imaging the high-ionic strength plume and in defining the transition zone between saprolite and bedrock zones that appears to have a significant influence on contaminant transport. The geophysical data were used to help select the location and depth of investigation for field research plots. Drilling, borehole geophysics, and ground water sampling were used to verify the surface geophysical studies. PMID- 15819939 TI - Long-term changes in ground water chemistry at a phytoremediation demonstration site. AB - A field-scale demonstration project was conducted to evaluate the capability of eastern cottonwood trees (Populus deltoides) to attenuate trichloroethene (TCE) contamination of ground water. By the middle of the sixth growing season, trees planted where depth to water was <3 m delivered enough dissolved organic carbon to the underlying aquifer to lower dissolved oxygen concentrations, to create iron-reducing conditions along the plume centerline and sulfate-reducing or methanogenic conditions in localized areas, and to initiate in situ reductive dechlorination of TCE. Apparent biodegradation rate constants for TCE along the centerline of the plume beneath the phytoremediation system increased from 0.0002/d to 0.02/d during the first six growing seasons. The corresponding increase in natural attenuation capacity of the aquifer along the plume centerline, from 0.0004/m to 0.024/m, is associated with a potential decrease in plume-stabilization distance from 9680 to 160 m. Demonstration results provide insight into the amount of vegetation and time that may be needed to achieve cleanup objectives at the field scale. PMID- 15819940 TI - Oxidation of fugitive methane in ground water linked to bacterial sulfate reduction. AB - When fugitive methane migrates upward along boreholes of oil and gas wells, it may migrate into shallow ground water or pass through overlying soil to the atmosphere. Prior to this study, there was little information on the fate of fugitive methane that migrates into ground water. In a field study near Lloydminster, Alberta, Canada, we found hydrogeochemical evidence that fugitive methane from an oil well migrated into a shallow aquifer but has been attenuated by dissimilatory bacterial sulfate reduction at low temperature ( approximately 5 degrees C) under anaerobic conditions. Evidence includes spatial and temporal trends in concentrations of methane and sulfate in ground water and associated trends in concentrations of bicarbonate and sulfide. Within 10 m of the oil well, sulfate concentrations were low, and sulfate was enriched in both 34S and 18O. Sulfate concentrations had a strong positive correlation with delta13C values of bicarbonate, and sulfide was depleted in 34S compared to sulfate. These data indicate that bacterial sulfate reduction occurred near the production well. Near the oil well, elevated concentrations of bicarbonate were observed, and the bicarbonate was depleted in 13C. Modeling indicates that the main source of this excess 13C-depleted bicarbonate is oxidized methane. In concert with the sulfate concentration and isotope data, these results support an interpretation that in situ bacterial oxidation of methane has occurred, linked to bacterial sulfate reduction. Bacterial sulfate reduction may play a major role in bioattenuation of fugitive natural gas in ground water in western Canada. PMID- 15819941 TI - Aquifer vulnerability assessment to heavy metals using ordinal logistic regression. AB - A methodology using ordinal logistic regression is proposed to predict the probability of occurrence of heavy metals in ground water. The predicted probabilities are defined with reference to the background concentration and the maximum contaminant level. The model is able to predict the occurrence due to different influencing variables such as the land use, soil hydrologic group (SHG), and surface elevation. The methodology was applied to the Sumas-Blaine Aquifer located in Washington State to predict the occurrence of five heavy metals. The influencing variables considered were (1) SHG; (2) land use; (3) elevation; (4) clay content; (5) hydraulic conductivity; and (6) well depth. The predicted probabilities were in agreement with the observed probabilities under existing conditions. The results showed that aquifer vulnerability to each heavy metal was related to different sets of influencing variables. However, all heavy metals had a strong influence from land use and SHG. The model results also provided good insight into the influence of various hydrogeochemical factors and land uses on the presence of each heavy metal. A simple economic analysis was proposed and demonstrated to evaluate the cost effects of changing the land use on heavy metal occurrence. PMID- 15819942 TI - Spatial weighting functions: transient hydraulic tests and heterogeneous media. AB - To improve understanding of property measurements in heterogeneous media, an energy-based weighting function concept is developed. In (assumed) homogeneous media, the instrument spatial weighting function (ISWF) depends only on the energy dissipation distribution set up by the measurement procedure and it reduces to simply inverse sample volume (uniform weighting) for 1-D parallel flow case (ideal permeameter). For 1-D transient flow in homogeneous media, such as with slug tests, the ISWF varies with position and time, with 95% of the total weighting contained within 115 well radii, even late in the test. In the heterogeneous case, the determination of the ISWF is connected to the problem of determining an equivalent hydraulic conductivity (K), where the criterion for equivalence is based on equal energy dissipation rate rather than equal volume discharge. The discharge-based equivalent K (K(E)) and the energy-based equivalent K in heterogeneous media (K(eh)) are not equal in general, with K(eh) typically above the nodal arithmetic mean K. The possibly more fundamental problem is that as one makes K measurements in heterogeneous media at different locations or on different cores of heterogeneous materials, the ISWF will be heterogeneity dependent, implying that the averaging process resulting in the equivalent K value also varies with position. If the testing procedure is transient, then the averaging process varies with time. This suggests a fundamental ambiguity in the interpretation of hydraulic conductivity measurements in heterogeneous media that may impact how we approach modeling and prediction in a practical sense (Molz 2003). Further research is suggested. PMID- 15819943 TI - High-resolution slug testing. AB - The hydraulic conductivity (K) variation has important ramifications for ground water flow and the transport of contaminants in ground water. The delineation of the nature of that variation can be critical to complete characterization of a site and the planning of effective and efficient remedial measures. Site-specific features (such as high-conductivity zones) need to be quantified. Our alluvial field site in the Kansas River valley exhibits spatial variability, very high conductivities, and nonlinear behavior for slug tests in the sand and gravel aquifer. High-resolution, multilevel slug tests have been performed in a number of wells that are fully screened. A general nonlinear model based on the Navier Stokes equation, nonlinear frictional loss, non-Darcian flow, acceleration effects, radius changes in the wellbore, and a Hvorslev model for the aquifer has been used to analyze the data, employing an automated processing system that runs within the Excel spreadsheet program. It is concluded that slug tests can provide the necessary data to identify the nature of both horizontal and vertical K variation in an aquifer and that improved delineation or higher resolution of K structure is possible with shorter test intervals. The gradation into zones of higher conductivity is sharper than seen previously, and the maximum conductivity observed is greater than previously measured. However, data from this project indicate that well development, the presence of fines, and the antecedent history of the well are important interrelated factors in regard to slug-test response and can prevent obtaining consistent results in some cases. PMID- 15819944 TI - A neural network model for predicting aquifer water level elevations. AB - Artificial neural networks (ANNs) were developed for accurately predicting potentiometric surface elevations (monitoring well water level elevations) in a semiconfined glacial sand and gravel aquifer under variable state, pumping extraction, and climate conditions. ANNs "learn" the system behavior of interest by processing representative data patterns through a mathematical structure analogous to the human brain. In this study, the ANNs used the initial water level measurements, production well extractions, and climate conditions to predict the final water level elevations 30 d into the future at two monitoring wells. A sensitivity analysis was conducted with the ANNs that quantified the importance of the various input predictor variables on final water level elevations. Unlike traditional physical-based models, ANNs do not require explicit characterization of the physical system and related physical data. Accordingly, ANN predictions were made on the basis of more easily quantifiable, measured variables, rather than physical model input parameters and conditions. This study demonstrates that ANNs can provide both excellent prediction capability and valuable sensitivity analyses, which can result in more appropriate ground water management strategies. PMID- 15819945 TI - Empirical relationships for estimating stream depletion by a well pumping near a gaining stream. AB - Siting wells near streams requires an accurate estimate of the quantity of water derived from the river due to pumping. A number of hydrogeological and hydraulic parameters influence this value. This study estimates stream depletion under steady-state conditions for a variety of hydrogeological systems. A finite differences model was used to analyze several hydrogeological situations, and for each of these the stream depletion was estimated using an advective transport method. An empirical equation for stream depletion was obtained for the case of a stream that partially penetrates the aquifer and a pumping well that is screened over a portion of the aquifer. The derived equation, which is valid for both isotropic and anisotropic conditions, expresses stream depletion as a function of the unit inflow to the river, the discharge of the pumping well, the well screen length, the distance between the river and pumping well, the wetted perimeter, and a new parameter called "overlap," which is defined to be the distance between the riverbed and the top of well screen. The overlap parameter makes it possible to consider indirectly the vertical component of flow, which is accentuated when the well is screened below the streambed. The formula proposed here should be useful in deciding where to locate a pumping well and to decide the appropriate length of its screen. PMID- 15819946 TI - Correlating fracture trends and hydraulic head using semivariogram cloud analysis. AB - Fracture trends (defined as kilometer-scale linear features interpolated between field observations of fractures along their strikes) often have a dominant orientation. Finding a correlation between this orientation and hydraulic data could shed light on their hydraulic influence. A significant correlation between head residuals from first-order regional drift and the orientation of 2- to 4-km long fracture trends was found in a study site in the Negev, Israel, using the semivariogram cloud analysis. Correlation of head residuals rather than the head itself implies that the orientation of the fracture trends controls the anisotropy and heterogeneity at this scale, mainly because the fracture trends define the orientation of blocks, which differ in their hydraulic properties. Preferential transmissive pathways are probably shorter than the full extent of the fracture trends, causing a relatively high head difference along the trends on the 2- to 4-km scale. Fracture trend density and additional data from short range hydraulic tests helped characterize two blocks separated by a fault zone. The identification of hydraulic features on a kilometer scale is necessary for better modeling of regional ground water flow and transport. Hydraulic tests at this scale are not feasible, thereby rendering combined analyses of head and structural data, such as the one presented here, essential. PMID- 15819947 TI - Transmissivity variations in mudstones. AB - Many people in sub-Saharan Africa have to rely on meager water resources within mudstones for their only water supply. Although mudstones have been extensively researched for their low permeability behavior, little research has been undertaken to examine their ability to provide sustainable water supplies. To investigate the factors controlling the occurrence of usable ground water in mudstone environments, an area of Cretaceous mudstones in southeastern Nigeria was studied over a 3 yr period. Transmissivity (T) variations in a range of mudstone environments were studied. The investigations demonstrate that within the top 40 m of mudstones, transmissivity can be sufficient to develop village water supplies (T > 1 m2/d). Transmissivity is controlled by two factors: low grade metamorphism and the presence of other, subordinate, lithologies within the mudstones. Largely unaltered mudstones (early diagenetic zone), comprising mainly smectite clays, are mostly unfractured and have a low T of < 0.1 m2/d. Mudstones that have undergone limited metamorphism (late diagenetic zone) comprise mixed layered illite/smectite clays, and ground water is found in widely spaced fracture zones (T > 1 m2/d in large fracture zones; T < 0.1 m2/d away from fracture zones). Mudstones that have been further altered and approach the anchizone comprise illite clays, are pervasively fractured, and have the highest transmissivity values (T > 4 m2/d). Dolerite intrusions in unaltered, smectitic mudstones are highly fractured with transmissivity in the range of 1 < T < 60 m2/d. Thin limestone and sandstone layers can also enhance transmissivity sufficiently to provide community water supplies. PMID- 15819948 TI - Improved water table dynamics in MODFLOW. AB - The standard formulation of a block-centered finite-difference model, such as MODFLOW, uses the center of the cell as the location of a cell node. Simulations of a dynamic water table can be improved if the node of a cell containing the water table is located at the water table rather than at the center of the cell. The LPF package of MOD-FLOW-2000 was changed to position a cell's node at the water table in convertible cells with a water table. Improved accuracy in the upper regions of an unconfined aquifer is demonstrated for pumping from a partially penetrating well. The change introduces a nonlinearity into the solution of the flow equations that results in slightly slower convergence of the flow solution, 7% slower in the presented demonstration. Accuracy of simulations is improved where vertical flow is dominated by a moving water table, but not when a large water table gradient dominates over the water table movement. PMID- 15819949 TI - An improved approach for assigning pumping rates to heterogeneous aquifer models. AB - A common assumption of ground water models formulated using a block-centered finite-difference method is that a well is located at the center of a cell regardless of its actual location. Due to this assumption, errors are introduced in the spatial distribution of simulated heads. This paper presents an alternative approach for assigning the pumping rates of wells that are located off cell centers. This approach consists of assigning the pumping rate not only to the cell in which the well is located but also to adjacent cells, taking into account the length of the well screen, the hydraulic conductivity, and the distance from the well to the center of its cell. The advantage of this alternative approach over the conventional one is illustrated with a test problem of a synthetic aquifer. Statistical measures of error indicate a much better model fit when pumping rates of wells are distributed over several cells. PMID- 15819950 TI - Approximate solution for solute transport during spherical-flow push-pull tests. AB - An approximate analytical solution to the advection-dispersion equation was derived to describe solute transport during spherical-flow conditions in single well push-pull tests. The spherical-flow case may be applicable to aquifer tests conducted in packed intervals or partially penetrating wells. Using results of two-dimensional numerical simulations, we briefly illustrate the applicability of the derived spherical-flow solution and provide a comparison with its cylindrical flow counterpart. Good agreement between simulated extraction-phase breakthrough curves and the spherical-flow solution was found when the length of the injection/extraction region was small compared to both aquifer thickness and maximum solute frontal position at the end of the injection phase. On the other hand, discrepancies between simulated breakthrough curves and the spherical-flow solution increased with increasing anisotropy in hydraulic conductivities. Several inherent limitations embedded in its derivation such as assumptions of isotropy and homogeneity warrant the cautious use of the spherical-flow solution. PMID- 15819951 TI - Hydrogeologic unit flow characterization using transition probability geostatistics. AB - This paper describes a technique for applying the transition probability geostatistics method for stochastic simulation to a MODFLOW model. Transition probability geostatistics has some advantages over traditional indicator kriging methods including a simpler and more intuitive framework for interpreting geologic relationships and the ability to simulate juxtapositional tendencies such as fining upward sequences. The indicator arrays generated by the transition probability simulation are converted to layer elevation and thickness arrays for use with the new Hydrogeologic Unit Flow package in MODFLOW 2000. This makes it possible to preserve complex heterogeneity while using reasonably sized grids and/or grids with nonuniform cell thicknesses. PMID- 15819952 TI - Ultrastructural and rheological properties of bovine vaginal fluid and its relation to sperm motility and fertilization: a review. AB - The tubular genital organs of the mammalian female reproductive tract provide different luminal microenvironments that can act as an aid or a barrier to gamete transport and the first steps of embryonic development. The mechanism of sperm transport through the genital tract has not been fully established but, first of all, sperm cells encounter and need to overcome mucous-like secretions to reach the site where fertilization occurs. One of these secretions, vaginal fluid, is a complex biological product, which is mainly derived from cervical mucus. In this review, we focus on recent studies that describe the relationships between the ultrastructural and rheological properties of bovine vaginal fluid and sperm motion. In the light of this, we propose bovine vaginal fluid as a morphofunctional model for the study of sperm transport in mammals. PMID- 15819953 TI - Number of follicular waves and their effect on pregnancy rate in the cow. AB - In the study, the relationship of follicular growth waves, oestradiol and pregnancy rates were investigated during oestrus cycle in cows. A total of 22, Brown Swiss cows (3-5 years old) were used for the study. The ovaries of animals were examined from sixth day of cycle to next oestrus by ultrasound. The follicles that were present in the ovarium were recorded. Follicular growth was observed every day by means of ultrasound examination. The blood samples were taken for analysis of oestradiol simultaneously with ultrasound examinations. The oestrus animals were inseminated. Each animal that inseminated was examined by ultrasound on day 28 after insemination for pregnancy diagnosis. Two follicular growth waves were observed in nine of 22 (40.9%), three waves in 13 of 22 (59.1%) animals. The oestradiol was found in the same concentration but in different release patterns between two and three waves animals. Pregnancy rate in cows with three and two follicular waves did not differ. In conclusion, emergence of three waves of follicular growth was higher in Brown Swiss cows, the analysis of oestradiol could be used for determination of the wave numbers and the animals with different waves may have had the same pregnancy rates. PMID- 15819954 TI - Effect of alpha-tocopherol and ascorbic acid on bovine oocyte in vitro maturation. AB - In vitro culture results in higher oxygen concentrations than in vivo environments, leading to an increased level of reactive oxygen species (ROS) that cause lipid peroxidation of cellular membranes. Alpha-tocopherol (active form of vitamin E) is an antioxidant that protects mammalian cells against lipid peroxidation, which is regenerated by ascorbic acid. The aim of this study was to determine the effect of the addition of alpha-tocopherol and/or ascorbic acid to the maturation medium on bovine oocyte in vitro maturation (IVM) and subsequently on in vitro fertilization (IVF) and embryo development. Cumulus-oocyte complexes (COCs) were matured in Medium 199 (control), and with the addition of alpha tocopherol and/or ascorbic acid. The concentration of alpha-tocopherol in COCs was determined by high-performance liquid chromatography (HPLC). IVF and in vitro culture (IVC) were carried out in modified synthetic oviductal fluid (mSOF). The quantity of alpha-tocopherol naturally present in COCs diminished by half during IVM (p < 0.05), although in the presence of ascorbic acid it remained constant. A greater amount of alpha-tocopherol was detected in COCs matured in medium supplemented with this antioxidant (p < 0.05), but the addition of alpha tocopherol plus ascorbic acid maintained higher levels of alpha-tocopherol (p < 0.05). Significant differences were not observed in the percentages of nuclear maturation and fertilization among different treatments. The presence of alpha tocopherol or ascorbic acid in the maturation medium failed to modify the percentage of blastocysts obtained, unlike the addition of both antioxidants when a significant decrease was observed (p < 0.05). Absorbic acid maintained the antioxidant capacity of the alpha-tocopherol incorporated to COC membranes during IVM. The active form of vitamin E during maturation impaired the acquisition of oocyte developmental competence. PMID- 15819955 TI - Preliminary evaluation of a unique freezing technology for bovine spermatozoa cryopreservation. AB - Artificial insemination (AI) and semen cryopreservation has significantly improved the breeding potential of male animals. However, current freezing techniques commonly result in reduced semen quality. Ten years ago, a unique freezing technology (UFT) was developed for the freezing of foodstuffs and other materials. Previous work from this laboratory has demonstrated the UFT to be a superior method of freezing for a number of cell types. In a preliminary study, the UFT was compared with the conventional freezing methodology of bovine semen. Semen samples were collected from an angus (Bull A) and a gelbivich bull (Bull B), prepared using a conventional bovine cryoprotectant, and frozen in the UFT or in liquid nitrogen (LN) mist. The samples were stored in LN before being thawed and assessed for the semen parameters of motility and forward progression. Preliminary results suggest the UFT is equivalent to current techniques in the cryopreservation and recovery of bovine semen, and with modification, possibly a superior technique for semen freezing. Further studies using larger sample populations, and using a CASA system to evaluate motility, forward progression and linearity are merited. PMID- 15819956 TI - The effect of oestradiol, progesterone and heparin on bovine spermatozoa function after thawing. AB - The present experiment was designed to determine the effects of various biologically active substances, such as oestradiol (OE), progesterone (P4) and heparin (Hep) alone or in combination on sperm plasma membrane scrambling, capacitation and acrosome reaction (AR) of post-thaw bovine spermatozoa. Spermatozoa were incubated for 180 min in capacitation medium supplemented with (i) 1 mug/ml OE; (ii) 1 mug/ml P4; (iii) 1 mug/ml OE and 1 mug/ml P4; (iv) 1 mug/ml OE and 5 mug/ml Hep; (v) 1 mug/ml P4 and 5 mug/ml Hep; (vi) 1 mug/ml OE, 1 mug/ml P4 and 5 mug/ml Hep. At predetermined time intervals aliquots were taken to assess sperm plasma membrane scrambling, or capacitation (AR induced by lysophosphatidylcholine) in spermatozoa. The second experiment was aimed to study the effects of OE, P4 and OE/P4 as potential inducers of AR in Hep-capacitated spermatozoa. Plasma membrane scrambling was assessed by a flow cytometer, using Merocyanine staining. Acrosomal status and viability of spermatozoa were evaluated under epifluorescence microscope with Ethidium homodimer-1/peanut agglutinin fluorescein isothiocyanate staining method (EthD-1/PNA-FITC). The results show that OE, P4 and a combination of OE/P4 at concentrations used did not affect sperm viability. Heparin significantly (p < 0.001) increased sperm plasma membrane scrambling of OE and P4-treated spermatozoa. P4 significantly affected the rate of sperm capacitation (p < 0.001) and AR (p < 0.05), but OE expressed membrane-stabilizing properties (p < 0.05). It can be concluded that in frozen-thawed bovine spermatozoa OE presents plasma membrane stabilizing properties that can be abolished by Hep, but not by P4. Progesterone possesses capacitating and AR-inducing properties in frozen-thawed bovine spermatozoa that can be alleviated by OE. PMID- 15819957 TI - Responsiveness to progestagen-eCG-cloprostenol treatment in goat food restricted for long period and refed. AB - For 6 months, 10 adult Saanen crossbred goats were fed undernutrition diet (70% maintenance), and finally five goats were refed for 6 weeks with 150% maintenance. In all animals oestrus was synchronized using 45 mg FGA vaginal sponge for 11 days, 300 IU eCG and 50 microg cloprostenol 48 h prior to sponge removal. From oestrus onset, during a 24-h period, blood samples were collected for oestradiol and NEFA assay. Ovulation was verified by laparoscopy 3 days after sponge removal. Body mass loss was 18.62 +/- 3.03% of initial weight and in refed goats body weight recovery was 90.63 +/- 3.56%. NEFA level was higher in restricted goats (p < 0.05). Fifty per cent of underfed goats (2/4) and all refed goats (4/4) exhibited oestrus and ovulation. Significant relationship (p < 0.05) was found between weight loss and the interval sponge removal-oestrus onset (r = 0.91) or ovulation rate (r = 0.70). Only in the refed group was the ovulation rate related to the oestradiol amount (r = 0.99) (p < 0.05). Collectively results showed that a short period of improved feeding re-established the responsiveness of oestrus synchronization in chronically fasted goats. PMID- 15819958 TI - Apoptosis in the antral follicles of swamp buffalo and cattle ovary: TUNEL and caspase-3 histochemistry. AB - The present study was carried out to investigate the pattern of apoptosis in the healthy antral and atretic follicles of Philippine swamp buffaloes (BU) in comparison with Holstein-Friesian (HF) cows. Paraffin sections of healthy follicles and various stages of atretic follicles were stained using the terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine triphosphates (dUTP) nick end-labelling (TUNEL) method to detect DNA fragmentation and cleaved caspase-3 antibody to detect cells committed to undergo apoptosis. Five equidistant areas of a follicle were counted for the presence of TUNEL- and caspase-3-positive cells. Healthy follicles of BU and HF contained no TUNEL-positive cells in the granulosa and theca layer but showed some caspase-3 positivity. The granulosa layer of advanced atretic follicles showed a significantly higher frequency of caspase-3 positivity than the healthy and early atretic follicles in both breeds. The frequency of caspase-3-positive cells of BU was significantly higher than HF in the granulosa layer of healthy, early atretic and advanced atretic follicles. In the theca interna layer, BU and HF showed a significantly lower and higher frequency of TUNEL-positive cells in the late atretic follicles compared with advanced atretic follicle, respectively. However, the frequency of caspase-3-positive cells of both BU and HF in the late atretic follicles was significantly higher than the advanced atretic follicles in the theca interna layer. These results indicate that caspase-3 and DNA fragmentation is involved in the buffalo ovarian apoptotic process. PMID- 15819959 TI - Acute insulin-induced hypoglycaemia does not alter IGF-1 and LH release in cyclic mares. AB - Lactation in the mare is associated with changes in the release of metabolic as well as reproductive hormones. Plasma glucose concentration is constantly reduced in lactating compared with non-lactating mares. Several metabolic signals have been proposed to link nutrition and somatic metabolism with reproductive function. The following experiment was performed to study the effect of acute hypoglycaemia on the release of insulin-like growth factor-1 (IGF-1) and luteinizing hormone (LH) in cyclic mares. Different doses of insulin (0.1 and 0.2 IU/kg body weight) were given to induce a decrease in plasma glucose concentration, as existent in lactating mares. All horses treated with insulin developed a hypoglycaemia over a time period of nearly 10 h. The IGF-1 and LH were analysed before and after insulin administration. At no point of time, a significant difference between the two insulin treatments and the control treatment was observed. Therefore, the hypoglycaemic horse is apparently able to provide the brain with sufficient glucose. Short-term hypoglycaemia does not affect the hypothalamo-pituitary-ovarian axis, and concentrations of IGF-1 and LH remained stable during insulin-induced hypoglycaemia. An acute change in plasma glucose concentration is thus not or at least not the only metabolic signal that links nutrition and somatic metabolism with reproductive function in the horse mare. PMID- 15819960 TI - Endometrial oxytocin receptor concentration and activity in prepubertal ewe lambs. AB - We have measured endometrial oxytocin receptor concentrations during prepuberty in ewe lambs, and have investigated the effect of progesterone on the activity of these receptors. In the first study, oxytocin receptor concentrations were undetectable in 2-week prenatal lambs but had increased immediately following birth and were then maintained throughout prepubertal life. Despite the presence of oxytocin receptors animals showed no prostaglandin F(2alpha) (PGF(2alpha)) release in response to exogenous oxytocin challenge at either 3 or 5 months of age. In a second study in 4-month-old ewe lambs treatment with exogenous progesterone resulted in the appearance of PGF(2alpha) release in response to oxytocin after 10 days of treatment. Thus, during the prepubertal life, ewe lambs possess the prerequisites of a luteolytic mechanism in that they have a dormant population of oxytocin receptors in which progesterone can induce oxytocin stimulated PGF(2alpha) release. PMID- 15819961 TI - Intracytoplasmic glutathione levels in heifer oocytes cultured in different maturation media and its effect on embryo development. AB - The present study was carried out to study the effect of different maturation media on embryo development of heifer oocytes and on their glutathione (GSH) synthesis during in vitro maturation (IVM). Immature heifer oocytes were matured in parallel in one of four maturation media: (i) Tissue Culture Medium (TCM)-199 supplemented with 10 ng/ml of epidermal growth factor (EGF); (i) TCM-199 supplemented with 10 ng/ml of EGF plus 1 microg/ml of FSH; (iii) TCM-199 supplemented with 10% of foetal bovine serum (FBS) and (iv) TCM-199 supplemented with 10% of FBS plus 1 microg/ml of FSH. Cow oocytes were used as control and were matured in TCM-199 supplemented with 10 ng/ml of EGF. No differences were observed in blastocyst rate among the different heifer oocyte groups (8.8, 7.5. 8.4 and 6.8%, respectively) however, the percentage of blastocysts obtained from cow oocytes was significantly higher (30%; p < 0.01) than those obtained from heifer oocytes. De novo GSH synthesis during oocyte maturation of heifer and cow oocytes was detected. No significant differences in intracytoplasmic GSH levels were observed among the experimental heifer oocyte groups or between heifer and cow oocytes both before and after IVM. In conclusion, the blastocyst yield obtained from heifer oocytes was lower than that from cow oocytes and this fact could not be explained by significant differences in intracytoplasmic GSH contents of oocytes before or after IVM. PMID- 15819962 TI - The proportion of beef bulls in Sweden with mature spermiograms at 11-13 months of age. AB - Semen samples collected postmortem from 142 yearling beef bulls (11-13 months old) of three different breeds (Charolais, Hereford and Simmental) were examined to evaluate the proportion of bulls with mature spermiograms. Before slaughter, testes and epididymides were clinically examined and scrotal circumferences were measured. Aliquots of the cauda epididymal contents taken postmortem were used for sperm morphology examination. Sperm head morphology was studied in dry smears stained with carbol-fuchsine. For each preparation, 500 spermatozoa were counted in each smear under light microscope (x 1000). The presence of proximal cytoplasmic droplets, abnormal acrosomes, detached heads and abnormalities of the midpiece and tail were recorded in wet preparations of formol-saline-fixed spermatozoa. For each preparation, 200 spermatozoa were counted in each preparation under a phase-contrast microscope (x 1000). The abnormalities were classified according to a classification system developed by Bane (1961). Morphological abnormalities were recorded as a percentage of the total number of counted spermatozoa. Criteria for a spermiogram to be considered mature included <15% abnormal heads and <15% proximal droplets. According to this definition approximately 48% (68 of 142) of the examined bulls were considered mature. The bulls in this study represent approximately one-fifth of the total amount of performance-tested beef bulls in Sweden during 5 years. Our results indicate that only less than half of the Swedish yearling beef bulls at the testing station appear to have a mature spermiogram at the time they are offered for breeding purposes. PMID- 15819963 TI - In vitro evaluation of in vivo fertilizing ability of frozen rabbit semen. AB - The present work studied different spermatozoa parameters and the ability of frozen rabbit spermatozoa to fertilize, in vitro, in vivo-matured oocytes, as a test to predict their in vivo fertility and prolificacy. Semen from rabbit bucks was frozen using two freezing protocols [in a freezer at -30 degrees C or in liquid nitrogen vapour (LNV)]. For the in vivo trial, females were inseminated with frozen-thawed spermatozoa. Oocytes used for in vitro testing were recovered 14 h after ovulation induction from donors and co-incubated with 2 x 10(6) frozen thawed spermatozoa during 4 h at 37 degrees C in Tyrode's medium under an atmosphere of 5% CO(2) in air with maximal humidity. After co-incubation period, presumptive zygotes were cultured in TCM199 supplemented with 20% foetal bovine serum (FBS), under the same conditions described above. Although no statistical differences were observed between freezing protocols in seminal parameters [motility rate: 40 and 35%, VCL: 35 and 46 microm/s, amplitude of lateral head displacement (ALH): 1.7 and 2.4 mum, for semen frozen at -30 degrees C and in LNV, respectively], significant differences were noted in the fertilizing ability in vivo and in vitro. Semen frozen at -30 degrees C showed the highest fertilizing ability in vitro (26.7% vs 6.2 and 8.7% for semen frozen at -30 degrees C, in LNV and fresh semen, respectively) and the lowest fertility rate in vivo (21.7% vs 64.2% and 70.6% for semen frozen at -30 degrees C, in LNV and fresh semen, respectively). Sperm frozen at -30 degrees C seemed to be more capacitated. PMID- 15819964 TI - Second oestrus synchronization and precocious embryo viability after puberty induction in gilts by the use of gonadotrophin treatment. AB - The use of exogenous gonadotrophins in puberty inducement and ovulation synchronization is a technique that has a positive influence on the management of swine. The purpose of this study was to verify the effects of a second gonadotrophin treatment [equine chorionic gonadotrophin (eCG) and luteinizing hormone (LH), intramuscularly (i.m.)] upon the second oestrus synchronization and fertility in gilts. Seventy-one NAIMA (Pen Ar Lan) gilts had their first oestrus (puberty inducement) induced by a hormonal treatment (eCG and LH). Then, they were randomly distributed into two treatments, with (T1) and without (C) gonadotrophin treatment at the second oestrus. The animals were fed with a single ration (16% of crude protein and 3286.73 kcal ME/kg), and timed artificial insemination performed at the second oestrus. Gilts were slaughtered for embryo recovery and ovary examination about 5 days after insemination. There was no evidence of a difference in the percentage of the second oestrus expression (T1 - 90.90% and C - 86.84%), the duration of the oestrus cycle (T1 - 19.62 +/- 0.82 days and C - 19.67 +/- 4.14 days), the percentage of follicular cysts (T1 - 15.15% and C - 18.42%) and number of ovulations (T1 - 14.60 +/- 5.7 and C - 13.23 +/- 4.8) between treatments (p > 0.05). However, the hormonal treatment (T1) showed minor oestrus dispersion and embryo viability (T1 - 8.4 +/- 5.6 and C - 11.2 +/- 4.6) (p < 0.05). These results indicate that the better synchronization and expression of the second oestrus when using gonadotrophins (eCG and LH) is followed by a lower embryo viability, which is probably the consequence of the heterogeneous follicle recruitment during the injection of eCG. PMID- 15819966 TI - Some characteristics of primary and secondary oestrous signs in high-producing dairy cows. AB - Shortened and weakened oestrous signs in dairy cows may cause a failure of oestrus detection and artificial insemination timing error leading to poor reproductive performance. The aims of this study were to investigate the duration of standing oestrus in high-producing dairy cows under a free stall system, to determine the duration of expression of secondary oestrous signs before and after standing oestrus (Expt 1) and to compare the duration and intensity of oestrus between cows and heifers (Expt 2). Cattle were checked for primary and secondary oestrous signs at an interval of 4 h. Heat detection aids were also used. In Expt 1, of 56 cows which were detected in oestrus, 36 cows (64.3%) showed standing oestrus and other 20 cows (36.6%) showed secondary oestrous signs only. Duration of the standing oestrus was 6.6 +/- 6.3 h on average (+/-SD), ranging between 2 and 32 h. The cows in standing oestrus showed secondary oestrous signs during a period from 9.6 +/- 8.1 h before onset of standing to 18.4 +/- 18.8 h after the end of standing oestrus. In the cows that did not show standing oestrus, expression of secondary oestrous signs were observed for 25.7 +/- 20.5 h, which was 7.5 h shorter than the average duration of oestrus in cows showing standing oestrus. In Expt 2, nine (82%) of the 11 lactating cows in oestrus showed standing, while all the 10 heifers exhibited standing oestrus. Average duration of standing oestrus was 6.4 +/- 4.3 h in cows and 6.2 +/- 3.9 h in heifers, respectively. It may be concluded that the duration of standing oestrus is substantially shortened in lactating dairy cows, and more than one-third of cows did not show standing oestrus. In cows showing standing oestrus, duration of expression of secondary oestrous signs before and after standing is not shortened. Duration of standing oestrus in heifers was as short as that in cows. PMID- 15819965 TI - Ovum pick-up in cycling and lactating postpartum swamp buffaloes (Bubalis bubalis). AB - The objective of this study was to evaluate the efficiency of Ovum Pick Up (OPU) in cycling (n = 5) and lactating, postpartum, swamp buffaloes (n = 6) with and without gonadotropin stimulation. The OPU was performed every two weeks in all groups of animals, for a total of six sessions. Thirty collections were performed in five cycling buffaloes and 36 collections in six lactating postpartum buffaloes. Buffaloes that received hormonal stimulation were given a total of 400 mg, follicle stimulating hormone (FSH), administered twice daily over 3 days in decreasing doses, together with 100 microg of GnRH, 24 h after the last FSH injection. Following a resting period of 1 month, the two groups of buffaloes, were subjected to the same OPU regimen, but without any hormonal treatment for an additional six OPU sessions. The number of aspirated follicles recorded from the hormonal stimulated, cycling animals and lactating, postpartum buffaloes was not significantly different, 7.2 +/- 3.7 and 9.0 +/- 3.2, respectively (p > 0.05). Recovered oocytes collected from the two groups of hormonally stimulated animals were also not statistically different: 3.7 +/- 2.7 in the cycling and 5.9 +/- 3.5 in the lactating postpartum group (p > 0.05). In the two groups of buffaloes not receiving hormonal stimulation, the number of aspirated follicles was not significantly different: 2.1 +/- 1.4 and 1.4 +/- 0.7 in cycling and lactating postpartum buffaloes respectively (p > 0.05). Recovered oocytes in the non treated groups were also similar: 1.4 +/- 1.3 vs 0.7 +/- 0.8 in cycling and lactating buffaloes (p > 0.05). Among stimulated buffaloes, most aspirated follicles were small in size (< or =5 mm), whereas they were mostly medium and large sizes in the non-treated buffaloes. The oocyte recovery rate in both the groups, cycling and lactating postpartum, were 51.6% and 69.5% in stimulated groups and 55.0% and 53.1% in non-stimulated groups (p > 0.05). The majority of recovered oocytes were single- and multi-layered, and the number was greater in the cycling than in the lactating, postpartum buffaloes. The number and quality of recovered oocytes was similar in all groups of buffaloes whether they were received or did not receive hormonal stimulation. Moreover no difference was found in multi- and single-layered oocytes between cycling and lactating, postpartum buffaloes. In conclusion, OPU can be performed successfully in swamp buffalo in different reproductive status and FSH administration was shown to increase the number of aspirated oocytes in both cycling and lactating, postpartum buffaloes. PMID- 15819967 TI - Free oestrone in adipose tissue and its relation to androstenone and skatole in entire male pigs. AB - Relationship between free oestrone and boar taint compounds in adipose tissue were studied in two groups of entire male pigs of different breeds. Group A consisted of 33 entire crossbred male pigs (dam Yorkshire and sire backcross Yorkshire x Wild Boar, generation seven). Group B consisted of 194 entire male pigs of crossbreeds between Swedish Hampshire (H) and Finnish Landrace (L), LH x H, H x LH, LH x LH (dam x sire). The measurements of free oestrone in adipose tissue were performed with a new method developed and validated in our laboratories. The standard curve was linear for concentrations of free oestrone ranging from 0.13 to 5.10 ng/g. The method exhibited parallelism of results between serial dilutions and a mean recovery of 97 +/- 13.7%. Intra-assay variations for samples with concentrations of free oestrone from 0.67 to 2.08 ng/g were from 9.23 to 11.94%. Inter-assay variations for the samples with concentrations of free oestrone from 0.89 to 2.96 ng/g were from 3.78 to 10.11%. The levels of free oestrone in fat from group A were well correlated with fat levels of androstenone (r = 0.66; p < 0.001) and levels of oestrone sulphate in peripheral plasma collected at the same time as the fat (r = 0.74, p < 0.001). The levels of free oestrone in fat from group B were significantly correlated to fat levels of androstenone (r = 0.68, p < 0.001) and skatole (r = 0.29, p < 0.001). In group B, age-related differences in fat levels of free oestrone, androstenone and skatole were studied. Free oestrone and skatole levels increased simultaneously at the age of 22 week (p < 0.05 for both), and androstenone levels increased at the age of 26 week (p < 0.05). It was suggested that the levels of free oestrone in adipose tissue might be used for the evaluation of hormonal status of male pigs as an alternative to plasma levels of testicular hormones. The levels of free oestrone might be involved in the regulation of skatole levels in fat as indicated by both the simultaneous increases in skatole and free oestrone levels in fat and positive correlation between skatole and free oestrone. PMID- 15819968 TI - Impact of growth factors on in vitro development of caprine oocytes at pre-antral stage. AB - The objective of this study was to examine the effect of various growth factors such as the epidermal growth factor (EGF) and insulin-like growth factor-I (IGF I) either individually or in association, in the presence of follicle-stimulating hormone (FSH) on the in vitro growth and viability of caprine oocytes at pre antral stage. Pre-antral follicles were disassociated enzymatically and mechanically from pre-pubertal caprine ovaries after the animals were anaesthetically ovariectomized. Caprine pre-antral follicles in group 1, 2, 3 and 4 were cultured in growth culture medium, growth culture medium + EGF, growth culture medium + IGF-I and growth culture medium + IGF-I + EGF, respectively, for 9 days. The results indicated that EGF (50 mg/l) increased the survival rate of oocytes, but decreased the growth rate of oocytes; IGF-I (100 mg/l) effectively maintained the survival of oocytes and stimulated their growth; IGF-I (100 mg/l) and EGF (50 mg/l) in combination produced a higher effect on both the survival and the growth rate of oocytes than IGF-I or EGF alone. Conclusively, the growth factors can effectively maintain the survival of caprine oocytes from pre-antral follicles and regulate their growth in culture. EGF and IGF-I in association could synergically meliorate the culture system of caprine pre-antral follicles. PMID- 15819969 TI - Effect of the removal of cumulus cells on the nuclear maturation, fertilization and development of porcine oocytes. AB - The present study was conducted to investigate the effects of attachment of cumulus cells to porcine oocytes during the process of maturation and fertilization on the nuclear maturation, fertilization and subsequent development after in vitro fertilization (IVF). In the first experiment, the cumulus cells were removed from cumulus-oocyte complexes (COCs) at 0, 24 and 42 h after the onset of maturation culture and were then cultured until reaching 42 h of cultivation. In the second experiment, COCs were denuded as described in the first experiment, then fertilized and cultured for 7 days. As a control, cumulus cells were allowed to maintain attachment to the oocytes until the end of IVF. The proportion of oocytes reaching metaphase II significantly increased with the delay in the removal treatment of cumulus cells. The proportion of normal fertilization gradually increased with delay in the removal treatment of cumulus cells from COCs until the end of IVF. However, no significant difference in the proportion of normal fertilization was found between the 42-h and control groups. The removal treatment of cumulus cells in the 0- and 24-h group significantly (p < 0.05) decreased the proportion of cleaved embryos when compared with the control, and none of them developed to the blastocyst stage. The proportion of development to the blastocyst stage was significantly higher (p < 0.05) in the control group than in the 42-h group (18.1% vs 12.4%; p < 0.05). The present study indicates that the attachment of cumulus cells to the oocyte during maturation and fertilization is important to support oocyte nuclear maturation, fertilization and subsequent embryo development. Particularly, the attachment of cumulus cells to the oocyte during IVF promotes embryonic development. PMID- 15819970 TI - Effects of confluent, roscovitine treatment and serum starvation on the cell cycle synchronization of bovine foetal fibroblasts. AB - The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors. PMID- 15819971 TI - Expression of retinoid X receptor transcripts and their significance for developmental competence in in vitro-produced pre-implantation-stage bovine embryos. AB - Retinoid X receptors are frequently implicated in their role in embryonic development. However, there is no report regarding their expression in embryos with different qualities. In this study, expression pattern and levels of retinoid X receptor transcripts in different qualities and stages of in vitro produced pre-implantation bovine embryos were examined using real-time polymerase chain reaction (PCR) techniques. Four independent cDNA pools per sample were prepared from mRNAs isolated from different developmental-stage embryos, after classifying them to be of good and bad quality. Primers were also designed to amplify products with PCR. Based on analysis, except retinoid X receptor gamma (RXRG) transcripts, the remaining two types (alpha and beta forms) were expressed in all samples with significantly higher (p < or = 0.05) levels until the eight cell stage compared with the stages thereafter. However, the transcript levels for bad-quality embryos were consistently lower in all cases in comparison with good-quality embryos. RXRG was identified in 39% of the samples, but all in the pre-embryonic genome activation development stage. Therefore, strong expression patterns of these transcripts in earlier stages indicates their possible role during the maternal phase of embryo development, and the variations of copy number in embryos with different qualities substantiates their possible candidacy as potential quality markers. Moreover, identifying the transcript variations in different qualities and expression of RXRG at these stages is a novel input to the pre-implantation-stage knowledge. PMID- 15819972 TI - Bovine posterior pituitary extract stimulates prolactin release from the anterior pituitary gland in vitro and in vivo in cattle. AB - It has been reported that the posterior pituitary (PP) gland contains a potent, unknown prolactin (PRL)-releasing factor (PRF) in rats. PRFs are assumed to be produced in neurones located within the hypothalamus, and to be peptidergic in nature. However, little is known about PRFs in domestic animals. To characterize the PRF in the PP of domestic animals, the present study examined the PRL releasing activity of an acidic extract from bovine PP (bPP) in vitro and in vivo in cattle. First, the PRL-releasing effect of bPP extract was compared with that of PRL-releasing peptide (PrRP), and thyrotropin-releasing hormone (TRH) from cultured bovine anterior pituitary cells. The extract significantly increased PRL concentrations in the culture medium, at doses of 0.002 and 0.02 eq./ml (one eq. is the PP extract from one animal), compared with the control (p < 0.05). PrRP failed to stimulate the release of PRL. TRH significantly increased PRL concentrations in the culture medium, at doses from 10(-9) to 10(-7) M, compared with the control (p < 0.05). The rate of increase in the PRL concentration, by 0.02 eq./ml bPP extract, was significantly greater than that in TRH (p < 0.05). Secondly, plasma PRL responses to the intravenous (i.v.) injection of bPP extract (0.5 eq./head), PrRP [3.59 mug/kg body weight (BW)], TRH (1 mug/kg BW), and a dopamine receptor antagonist (sulpiride, 0.1 mg/kg BW), were examined in calves. PrRP failed to stimulate PRL release; however, plasma PRL increased immediately following the injection of bPP extract, TRH and sulpiride. The PRL-releasing effect of i.v. injections of TRH and sulpiride was more potent than that of bPP extract. Finally, plasma PRL responses to the intra-hypothalamic injection of bPP extract were examined in calves. The intra-hypothalamic infusion (arcuate nucleus) of 0.0625 eq./head of bPP extract strongly stimulated PRL release in calves (p < 0.05). The present results show that PP contains a physiologically potent PRF in cattle. PMID- 15819973 TI - Finite volume analysis of temperature effects induced by active MRI implants with cylindrical symmetry: 1. Properly working devices. AB - BACKGROUND: Active Magnetic Resonance Imaging implants are constructed as resonators tuned to the Larmor frequency of a magnetic resonance system with a specific field strength. The resonating circuit may be embedded into or added to the normal metallic implant structure. The resonators build inductively coupled wireless transmit and receive coils and can amplify the signal, normally decreased by eddy currents, inside metallic structures without affecting the rest of the spin ensemble. During magnetic resonance imaging the resonators generate heat, which is additional to the usual one described by the specific absorption rate. This induces temperature increases of the tissue around the circuit paths and inside the lumen of an active implant and may negatively influence patient safety. METHODS: This investigation provides an overview of the supplementary power absorbed by active implants with a cylindrical geometry, corresponding to vessel implants such as stents, stent grafts or vena cava filters. The knowledge of the overall absorbed power is used in a finite volume analysis to estimate temperature maps around different implant structures inside homogeneous tissue under worst-case assumptions. The "worst-case scenario" assumes thermal heat conduction without blood perfusion inside the tissue around the implant and mostly without any cooling due to blood flow inside vessels. RESULTS: The additional power loss of a resonator is proportional to the volume and the quality factor, as well as the field strength of the MRI system and the specific absorption rate of the applied sequence. For properly working devices the finite volume analysis showed only tolerable heating during MRI investigations in most cases. Only resonators transforming a few hundred mW into heat may reach temperature increases over 5 K. This requires resonators with volumes of several ten cubic centimeters, short inductor circuit paths with only a few 10 cm and a quality factor above ten. Using MR sequences, for which the MRI system manufacturer declares the highest specific absorption rate of 4 W/kg, vascular implants with a realistic construction, size and quality factor do not show temperature increases over a critical value of 5 K. CONCLUSION: The results show dangerous heating for the assumed "worst-case scenario" only for constructions not acceptable for vascular implants. Realistic devices are safe with respect to temperature increases. However, this investigation discusses only properly working devices. Ruptures or partial ruptures of the wires carrying the electric current of the resonance circuits or other defects can set up a power source inside an extremely small volume. The temperature maps around such possible "hot spots" should be analyzed in an additional investigation. PMID- 15819974 TI - Abacavir, efavirenz, didanosine, with or without hydroxyurea, in HIV-infected adults failing initial nucleoside/protease inhibitor-containing regimens. AB - BACKGROUND: Hydroxyurea (HU) is an immunomodulatory agent that has been documented to enhance the antiretroviral activity of nucleoside reverse transcriptase inhibitors, such as abacavir (ABC) and didanosine (ddI), and would be expected to improve virologic efficacy. METHODS: A 48-week, phase IV, multicenter, open-label, proof-of-concept clinical trial was conducted to evaluate second-line, protease inhibitor (PI)-sparing therapy with ABC/efavirenz (EFV)/ddI plus HU or without HU in HIV-infected subjects failing to achieve HIV-1 RNA < or = 400 copies/mL after > or = 16 weeks of treatment with lamivudine/zidovudine or lamivudine/stavudine, plus 1 or 2 PIs. Subjects were assigned to ABC (300 mg twice daily)/ EFV (600 mg once daily)/ ddI (400 mg once daily) plus HU (500 mg twice daily) (n = 30) or this regimen without HU (n = 24). RESULTS: Baseline mean HIV-1 RNA was 3.86 log10 copies/mL and CD4+ cell count was 345 cells/mm3. A similar percentage of subjects in the non-HU arm (58%) and HU arm (53%) completed the study. Intent-to-treat: missing = failure analysis showed no differences in proportions of subjects in the non-HU and HU arms achieving undetectable plasma HIV-1 RNA levels at week 24 (< 400 copies/mL: 58% [14/24] vs 57% [17/30], P = 0.899; < 50 copies/mL (50% [12/24] vs 47% [14/30], P = 0.780). Median change from baseline in CD4+ cell count in the non-HU and HU arms at week 48 was +114 cells/mm3 and -63 cells/mm3 (P = 0.007), respectively. Both regimens were generally well tolerated, although more subjects in the HU arm withdrew prematurely from the study due to adverse events (23% vs 4%). Four cases of possible ABC-related hypersensitivity were observed. CONCLUSION: ABC/EFV/ddI was an effective and well-tolerated second-line regimen for nucleoside/PI-experienced HIV-infected subjects. The addition of HU blunted the CD4+ cell response, did not appear to enhance antiviral activity, and resulted in more treatment-limiting adverse events. PMID- 15819975 TI - Does socioeconomic status affect mortality subsequent to hospital admission for community acquired pneumonia among older persons? AB - BACKGROUND: Low socioeconomic status has been associated with increased morbidity and mortality for various health conditions. The purpose of this study was twofold: to examine the mortality experience of older persons admitted to hospital with community acquired pneumonia and to test the hypothesis of whether an association exists between socioeconomic status and mortality subsequent to hospital admission for community-acquired pneumonia. METHODS: A population based retrospective cohort study was conducted including all older persons patients admitted to Ontario hospitals with community acquired pneumonia between April 1995 and March 2001. The main outcome measures were 30 day and 1 year mortality subsequent to hospital admission for community-acquired pneumonia. RESULTS: Socioeconomic status for each patient was imputed from median neighbourhood income. Multivariate analyses were undertaken to adjust for age, sex, co-morbid illness, hospital and physician characteristics. The study sample consisted of 60,457 people. Increasing age, male gender and high co-morbidity increased the risk for mortality at 30 days and one year. Female gender and having a family physician as attending physician reduced mortality risk. The adjusted odds of death after 30-days for the quintiles compared to the lowest income quintile (quintile 1) were 1.02 (95% CI: 0.95-1.09) for quintile 2, 1.04 (95% CI: 0.97 1.12) for quintile 3, 1.01 (95% CI: 0.94-1.08) for quintile 4 and 1.03 (95% CI: 0.96-1.12) for the highest income quintile (quintile 5). For 1 year mortality, compared to the lowest income quintile the adjusted odds ratios were 1.01 (95% CI: 0.96-1.06) for quintile 2, 0.99 (95% CI: 0.94-1.04) for quintile 3, 0.99 (95% CI: 0.93-1.05) for quintile 4 and 1.03 (95% CI: 0.97-1.10) for the highest income quintile. CONCLUSION: Socioeconomic status is not associated with mortality in the older persons from community-acquired pneumonia in Ontario, Canada. PMID- 15819977 TI - Monochorionic triamniotic triplet pregnancy with a co-triplet fetus discordant for congenital cystic adenomatoid malformation of the lung. AB - BACKGROUND: Spontaneous monochorionic triamniotic pregnancy is rare and is at increased risk for pregnancy complications. The presence of an anomalous fetus further complicates the management. CASE PRESENTATION: We present a case of monochorionic triamniotic triplet pregnancy diagnosed at 15 weeks of gestation with one fetus having developed a multicystic lung lesion, suggestive of congenital cystic adenomatoid malformation (CCAM). At 24 weeks, the largest cyst measured 10 mm in diameter. We managed the pregnancy conservatively and delivered three live male fetuses with birth weights 1560 g, 1580 g and 1590 g at 35 weeks of gestation. Two newborns were admitted to the neonatal intensive care unit with respiratory distress, the third one died due to sepsis 7 days postpartum. One of the newborns was discharged healthy at 24 days postpartum. The newborn with CCAM developed a pneumothorax on the right side, recovered after treatment, and was discharged after one month. Computerized tomography (CT) of the infant at 3 months demonstrated two cystic lesions in the middle lobe of the right lung measuring 25 mm and 15 mm. A repeat CT of the infant at 6 months showed a 30 mm solitary cystic mass. CONCLUSION: Monochorionic triamniotic triplet pregnancy with a co-triplet fetus discordant for CCAM, present rarely and can be managed conservatively. These findings may help in decision making and counselling of parents. PMID- 15819976 TI - A restraint molecular dynamics and simulated annealing approach for protein homology modeling utilizing mean angles. AB - BACKGROUND: We have developed the program PERMOL for semi-automated homology modeling of proteins. It is based on restrained molecular dynamics using a simulated annealing protocol in torsion angle space. As main restraints defining the optimal local geometry of the structure weighted mean dihedral angles and their standard deviations are used which are calculated with an algorithm described earlier by Doker et al. (1999, BBRC, 257, 348-350). The overall long range contacts are established via a small number of distance restraints between atoms involved in hydrogen bonds and backbone atoms of conserved residues. Employing the restraints generated by PERMOL three-dimensional structures are obtained using standard molecular dynamics programs such as DYANA or CNS. RESULTS: To test this modeling approach it has been used for predicting the structure of the histidine-containing phosphocarrier protein HPr from E. coli and the structure of the human peroxisome proliferator activated receptor gamma (Ppar gamma). The divergence between the modeled HPr and the previously determined X ray structure was comparable to the divergence between the X-ray structure and the published NMR structure. The modeled structure of Ppar gamma was also very close to the previously solved X-ray structure with an RMSD of 0.262 nm for the backbone atoms. CONCLUSION: In summary, we present a new method for homology modeling capable of producing high-quality structure models. An advantage of the method is that it can be used in combination with incomplete NMR data to obtain reasonable structure models in accordance with the experimental data. PMID- 15819978 TI - Quantification of collagen and proteoglycan deposition in a murine model of airway remodelling. AB - BACKGROUND: Sub-epithelial extracellular matrix deposition is a feature of asthmatic airway remodelling associated with severity of disease, decline in lung function and airway hyperresponsiveness. The composition of, and mechanisms leading to, this increase in subepithelial matrix, and its importance in the pathogenesis of asthma are unclear. This is partly due to limitations of the current models and techniques to assess airway remodelling. METHODS: In this study we used a modified murine model of ovalbumin sensitisation and challenge to reproduce features of airway remodelling, including a sustained increase in sub epithelial matrix deposition. In addition, we have established techniques to accurately and specifically measure changes in sub-epithelial matrix deposition, using histochemical and immunohistochemical staining in conjunction with digital image analysis, and applied these to the measurement of collagen and proteoglycans. RESULTS: 24 hours after final ovalbumin challenge, changes similar to those associated with acute asthma were observed, including inflammatory cell infiltration, epithelial cell shedding and goblet cell hyperplasia. Effects were restricted to the bronchial and peribronchial regions with parenchymal lung of ovalbumin sensitised and challenged mice appearing histologically normal. By 12 days, the acute inflammatory changes had largely resolved and increased sub epithelial staining for collagen and proteoglycans was observed. Quantitative digital image analysis confirmed the increased deposition of sub-epithelial collagen (33%, p < 0.01) and proteoglycans (32%, p < 0.05), including decorin (66%, p < 0.01). In addition, the increase in sub-epithelial collagen deposition was maintained for at least 28 days (48%, p < 0.001). CONCLUSION: This animal model reproduces many of the features of airway remodelling found in asthma and allows accurate and reproducible measurement of sub-epithelial extra-cellular matrix deposition. As far as we are aware, this is the first demonstration of increased sub-epithelial proteoglycan deposition in an animal model of airway remodelling. This model will be useful for measurement of other matrix components, as well as for assessment of the molecular mechanisms contributing to, and agents to modulate airway remodelling. PMID- 15819979 TI - Deduction of probable events of lateral gene transfer through comparison of phylogenetic trees by recursive consolidation and rearrangement. AB - BACKGROUND: When organismal phylogenies based on sequences of single marker genes are poorly resolved, a logical approach is to add more markers, on the assumption that weak but congruent phylogenetic signal will be reinforced in such multigene trees. Such approaches are valid only when the several markers indeed have identical phylogenies, an issue which many multigene methods (such as the use of concatenated gene sequences or the assembly of supertrees) do not directly address. Indeed, even when the true history is a mixture of vertical descent for some genes and lateral gene transfer (LGT) for others, such methods produce unique topologies. RESULTS: We have developed software that aims to extract evidence for vertical and lateral inheritance from a set of gene trees compared against an arbitrary reference tree. This evidence is then displayed as a synthesis showing support over the tree for vertical inheritance, overlaid with explicit lateral gene transfer (LGT) events inferred to have occurred over the history of the tree. Like splits-tree methods, one can thus identify nodes at which conflict occurs. Additionally one can make reasonable inferences about vertical and lateral signal, assigning putative donors and recipients. CONCLUSION: A tool such as ours can serve to explore the reticulated dimensionality of molecular evolution, by dissecting vertical and lateral inheritance at high resolution. By this, we mean that individual nodes can be examined not only for congruence, but also for coherence in light of LGT. We assert that our tools will facilitate the comparison of phylogenetic trees, and the interpretation of conflicting data. PMID- 15819980 TI - Location of DNA damage by charge exchanging repair enzymes: effects of cooperativity on location time. AB - BACKGROUND: How DNA repair enzymes find the relatively rare sites of damage is not known in great detail. Recent experiments and molecular data suggest that individual repair enzymes do not work independently of each other, but interact with each other through charges exchanged along the DNA. A damaged site in the DNA hinders this exchange. The hypothesis is that the charge exchange quickly liberates the repair enzymes from error-free stretches of DNA. In this way, the sites of damage are located more quickly; but how much more quickly is not known, nor is it known whether the charge exchange mechanism has other observable consequences. RESULTS: Here the size of the speed-up gained from this charge exchange mechanism is calculated and the characteristic length and time scales are identified. In particular, for Escherichia coli, I estimate the speed-up is 50000/N, where N is the number of repair enzymes participating in the charge exchange mechanism. Even though N is not exactly known, a speed-up of order 10 is not entirely unreasonable. Furthermore, upon over expression of all the repair enzymes, the location time only varies as N-1/2 and not as 1/N. CONCLUSION: The revolutionary hypothesis that DNA repair enzymes use charge exchange along DNA to locate damaged sites more efficiently is actually sound from a purely theoretical point of view. Furthermore, the predicted collective behavior of the location time is important in assessing the impact of stress-ful and radioactive environments on individual cell mutation rates. PMID- 15819981 TI - Mutation analysis of genes that control the G1/S cell cycle in melanoma: TP53, CDKN1A, CDKN2A, and CDKN2B. AB - BACKGROUND: The role of genes involved in the control of progression from the G1 to the S phase of the cell cycle in melanoma tumors in not fully known. The aim of our study was to analyse mutations in TP53, CDKN1A, CDKN2A, and CDKN2B genes in melanoma tumors and melanoma cell lines METHODS: We analysed 39 primary and metastatic melanomas and 9 melanoma cell lines by single-stranded conformational polymorphism (SSCP). RESULTS: The single-stranded technique showed heterozygous defects in the TP53 gene in 8 of 39 (20.5%) melanoma tumors: three new single point mutations in intronic sequences (introns 1 and 2) and exon 10, and three new single nucleotide polymorphisms located in introns 1 and 2 (C to T transition at position 11701 in intron 1; C insertion at position 11818 in intron 2; and C insertion at position 11875 in intron 2). One melanoma tumor exhibited two heterozygous alterations in the CDKN2A exon 1 one of which was novel (stop codon, and missense mutation). No defects were found in the remaining genes. CONCLUSION: These results suggest that these genes are involved in melanoma tumorigenesis, although they may be not the major targets. Other suppressor genes that may be informative of the mechanism of tumorigenesis in skin melanomas should be studied. PMID- 15819982 TI - The evolving role of the dynamic thermal analysis in the early detection of breast cancer. AB - It is now recognised that the breast exhibits a circadian rhythm which reflects its physiology. There is increasing evidence that rhythms associated with malignant cells proliferation are largely non-circadian and that a circadian to ultradian shift may be a general correlation to neoplasia.Cancer development appears to generate its own thermal signatures and the complexity of these signatures may be a reflection of its degree of development.The limitations of mammography as a screening modality especially in young women with dense breasts necessitated the development of novel and more effective screening strategies with a high sensitivity and specificity. Dynamic thermal analysis of the breast is a safe, non invasive approach that seems to be sensitive for the early detection of breast cancer.This article focuses on dynamic thermal analysis as an evolving method in breast cancer detection in pre-menopausal women with dense breast tissue. Prospective multi-centre trials are required to validate this promising modality in screening.The issue of false positives require further investigation using molecular genetic markers of malignancy and novel techniques such as mammary ductoscopy. PMID- 15819983 TI - Outcome of infections due to pandrug-resistant (PDR) Gram-negative bacteria. AB - BACKGROUND: The increasing problem of infections due to multidrug-resistant Gram negative bacteria has led to re-use of polymyxins in several countries. However, there are already clinical isolates of Gram-negative bacteria that are resistant to all available antibiotics, including polymyxins. METHODS: We present a case series of patients with infections due to pathogens resistant to all antimicrobial agents tested, including polymyxins. An isolate was defined as pandrug-resistant (PDR) if it exhibited resistance to all 7 anti-pseudomonal antimicrobial agents, i.e. antipseudomonal penicillins, cephalosporins, carbapenems, monobactams, quinolones, aminoglycosides, and polymyxins. RESULTS: Clinical cure of the infection due to pandrug-resistant (PDR) Gram-negative bacteria, namely Pseudomonas aeruginosa or Klebsiella pneumoniae was observed in 4 out of 6 patients with combination of colistin and beta lactam antibiotics. CONCLUSION: Colistin, in combination with beta lactam antibiotics, may be a useful agent for the management of pandrug-resistant Gram-negative bacterial infections. The re-use of polymyxins, an old class of antibiotics, should be done with caution in an attempt to delay the rate of development of pandrug-resistant Gram-negative bacterial infections. PMID- 15819984 TI - Deficiency in type 1 insulin-like growth factor receptor in mice protects against oxygen-induced lung injury. AB - BACKGROUND: Cellular responses to aging and oxidative stress are regulated by type 1 insulin-like growth factor receptor (IGF-1R). Oxidant injury, which is implicated in the pathophysiology of a number of respiratory diseases, acutely upregulates IGF-1R expression in the lung. This led us to suspect that reduction of IGF-1R levels in lung tissue could prevent deleterious effects of oxygen exposure. METHODS: Since IGF-1R null mutant mice die at birth from respiratory failure, we generated compound heterozygous mice harboring a hypomorphic (Igf 1rneo) and a knockout (Igf-1r-) receptor allele. These IGF-1Rneo/- mice, strongly deficient in IGF-1R, were subjected to hyperoxia and analyzed for survival time, ventilatory control, pulmonary histopathology, morphometry, lung edema and vascular permeability. RESULTS: Strikingly, after 72 h of exposure to 90% O2, IGF 1Rneo/- mice had a significantly better survival rate during recovery than IGF 1R+/+ mice (77% versus 53%, P < 0.05). The pulmonary injury was consistently, and significantly, milder in IGF-1Rneo/- mice which developed conspicuously less edema and vascular extravasation than controls. Also, hyperoxia-induced abnormal pattern of breathing which precipitated respiratory failure was elicited less frequently in the IGF-1Rneo/- mice. CONCLUSION: Together, these data demonstrate that a decrease in IGF-1R signaling in mice protects against oxidant-induced lung injury. PMID- 15819985 TI - Effective antiprotease-antibiotic treatment of experimental anthrax. AB - BACKGROUND: Inhalation anthrax is characterized by a systemic spread of the challenge agent, Bacillus anthracis. It causes severe damage, including multiple hemorrhagic lesions, to host tissues and organs. It is widely believed that anthrax lethal toxin secreted by proliferating bacteria is a major cause of death, however, the pathology of intoxication in experimental animals is drastically different from that found during the infectious process. In order to close a gap between our understanding of anthrax molecular pathology and the most prominent clinical features of the infectious process we undertook bioinformatic and experimental analyses of potential proteolytic virulence factors of B. anthracis distinct from lethal toxin. METHODS: Secreted proteins (other than lethal and edema toxins) produced by B. anthracis were tested for tissue-damaging activity and toxicity in mice. Chemical protease inhibitors and rabbit immune sera raised against B. anthracis proteases were used to treat mice challenged with B. anthracis (Sterne) spores. RESULTS: B. anthracis strain delta Ames (pXO1 , pXO2-) producing no lethal and edema toxins secrets a number of metalloprotease virulence factors upon cultivation under aerobic conditions, including those with hemorrhagic, caseinolytic and collagenolytic activities, belonging to M4 and M9 thermolysin and bacterial collagenase families, respectively. These factors are directly toxic to DBA/2 mice upon intratracheal administration at 0.5 mg/kg and higher doses. Chemical protease inhibitors (phosphoramidon and 1, 10 phenanthroline), as well as immune sera against M4 and M9 proteases of B. anthracis, were used to treat mice challenged with B. anthracis (Sterne) spores. These substances demonstrate a substantial protective efficacy in combination with ciprofloxacin therapy initiated as late as 48 h post spore challenge, compared to the antibiotic alone. CONCLUSION: Secreted proteolytic enzymes are important pathogenic factors of B. anthrasis, which can be considered as effective therapeutic targets in the development of anthrax treatment and prophylactic approaches complementing anti-lethal toxin therapy. PMID- 15819987 TI - Tissue Doppler echocardiographic quantification. Comparison to coronary angiography results in Acute Coronary Syndrome patients. AB - BACKGROUND: Multiples indices have been described using tissue Doppler imaging (DTI) capabilities. The aim of this study was to assess the capability of one or several regional DTI parameters in separating control from ischemic myocardium. METHODS: Twenty-eight patients with acute myocardial infarction were imaged within 24-hour following an emergent coronary angioplasty. Seventeen controls without any coronary artery or myocardial disease were also explored. Global and regional left ventricular functions were assessed. High frame rate color DTI cineloop recordings were made in apical 4 and 2-chamber for subsequent analysis. Peak velocity during isovolumic contraction time (IVC), ejection time, isovolumic relaxation (IVR) and filling time were measured at the mitral annulus and the basal, mid and apical segments of each of the walls studied as well as peak systolic displacement and peak of strain. RESULTS: DTI-analysis enabled us to discriminate between the 3 populations (controls, inferior and anterior AMI). Even in non-ischemic segments, velocities and displacements were reduced in the 2 AMI populations. Peak systolic displacement was the best parameter to discriminate controls from AMI groups (wall by wall, p was systematically < 0.01). The combination IVC + and IVR< 1 discriminated ischemic from non-ischemic segments with 82% sensitivity and 85% specificity. CONCLUSION: DTI-analysis appears to be valuable in ischemic heart disease assessment. Its clinical impact remains to be established. However this simple index might really help in intensive care unit routine practice. PMID- 15819988 TI - Bifrontal transcranial direct current stimulation slows reaction time in a working memory task. AB - BACKGROUND: Weak transcortical direct current stimulation (tDCS) applied to the cortex can shift the membrane potential of superficial neurons thereby modulating cortical excitability and activity. Here we test the possibility of modifying ongoing activity associated with working memory by tDCS. The concept of working memory applies to a system that is capable of transiently storing and manipulating information, as an integral part of the human memory system. We applied anodal and cathodal transcranial direct current (tDCS) stimulation (260 microA) bilaterally at fronto-cortical electrode sites on the scalp over 15 min repeatedly (15 sec-on/15 sec-off) as well as sham-tDCS while subjects performed a modified Sternberg task. RESULTS: Reaction time linearly increased with increasing set size. The slope of this increase was closely comparable for real and sham stimulation indicating that our real stimulation did not effect time required for memory scanning. However, reaction time was slowed during both anodal and cathodal stimulation as compared to placebo (p < 0.05) indicating that real stimulation hampered neuronal processing related to response selection and preparation. CONCLUSION: Intermittent tDCS over lateral prefrontal cortex during a working memory task impairs central nervous processing related to response selection and preparation. We conclude that this decrease in performance by our protocol of intermittent stimulation results from an interference mainly with the temporal dynamics of cortical processing as indexed by event-related sustained and oscillatory EEG activity such as theta. PMID- 15819986 TI - Interstitial lung disease in children -- genetic background and associated phenotypes. AB - Interstitial lung disease in children represents a group of rare chronic respiratory disorders. There is growing evidence that mutations in the surfactant protein C gene play a role in the pathogenesis of certain forms of pediatric interstitial lung disease. Recently, mutations in the ABCA3 transporter were found as an underlying cause of fatal respiratory failure in neonates without surfactant protein B deficiency. Especially in familiar cases or in children of consanguineous parents, genetic diagnosis provides an useful tool to identify the underlying etiology of interstitial lung disease. The aim of this review is to summarize and to describe in detail the clinical features of hereditary interstitial lung disease in children. The knowledge of gene variants and associated phenotypes is crucial to identify relevant patients in clinical practice. PMID- 15819989 TI - Shortest triplet clustering: reconstructing large phylogenies using representative sets. AB - BACKGROUND: Understanding the evolutionary relationships among species based on their genetic information is one of the primary objectives in phylogenetic analysis. Reconstructing phylogenies for large data sets is still a challenging task in Bioinformatics. RESULTS: We propose a new distance-based clustering method, the shortest triplet clustering algorithm (STC), to reconstruct phylogenies. The main idea is the introduction of a natural definition of so called k-representative sets. Based on k-representative sets, shortest triplets are reconstructed and serve as building blocks for the STC algorithm to agglomerate sequences for tree reconstruction in O(n2) time for n sequences. Simulations show that STC gives better topological accuracy than other tested methods that also build a first starting tree. STC appears as a very good method to start the tree reconstruction. However, all tested methods give similar results if balanced nearest neighbor interchange (BNNI) is applied as a post processing step. BNNI leads to an improvement in all instances. The program is available at http://www.bi.uni-duesseldorf.de/software/stc/. CONCLUSION: The results demonstrate that the new approach efficiently reconstructs phylogenies for large data sets. We found that BNNI boosts the topological accuracy of all methods including STC, therefore, one should use BNNI as a post-processing step to get better topological accuracy. PMID- 15819990 TI - Power and sample size calculations in the presence of phenotype errors for case/control genetic association studies. AB - BACKGROUND: Phenotype error causes reduction in power to detect genetic association. We present a quantification of phenotype error, also known as diagnostic error, on power and sample size calculations for case-control genetic association studies between a marker locus and a disease phenotype. We consider the classic Pearson chi-square test for independence as our test of genetic association. To determine asymptotic power analytically, we compute the distribution's non-centrality parameter, which is a function of the case and control sample sizes, genotype frequencies, disease prevalence, and phenotype misclassification probabilities. We derive the non-centrality parameter in the presence of phenotype errors and equivalent formulas for misclassification cost (the percentage increase in minimum sample size needed to maintain constant asymptotic power at a fixed significance level for each percentage increase in a given misclassification parameter). We use a linear Taylor Series approximation for the cost of phenotype misclassification to determine lower bounds for the relative costs of misclassifying a true affected (respectively, unaffected) as a control (respectively, case). Power is verified by computer simulation. RESULTS: Our major findings are that: (i) the median absolute difference between analytic power with our method and simulation power was 0.001 and the absolute difference was no larger than 0.011; (ii) as the disease prevalence approaches 0, the cost of misclassifying a unaffected as a case becomes infinitely large while the cost of misclassifying an affected as a control approaches 0. CONCLUSION: Our work enables researchers to specifically quantify power loss and minimum sample size requirements in the presence of phenotype errors, thereby allowing for more realistic study design. For most diseases of current interest, verifying that cases are correctly classified is of paramount importance. PMID- 15819991 TI - Diffusion and dissemination of evidence-based dietary strategies for the prevention of cancer. AB - OBJECTIVE: The purpose was to determine what strategies have been evaluated to disseminate cancer control interventions that promote the uptake of adult healthy diet? METHODS: A systematic review was conducted. Studies were identified by searching MEDLINE, PREMEDLINE, Cancer LIT, EMBASE/Excerpta Medica, PsycINFO, CINAHL, the Cochrane Database of Systematic Reviews, and reference lists and by contacting technical experts. English-language primary studies were selected if they evaluated the dissemination of healthy diet interventions in individuals, healthcare providers, or institutions. Studies of children or adolescents only were excluded. RESULTS: One hundred one articles were retrieved for full text screening. Nine reports of seven distinct studies were included; four were randomized trials, one was a cohort design and three were descriptive studies. Six studies were rated as methodologically weak, and one was rated as moderate. Studies were not meta-analyzed because of heterogeneity, low methodological quality, and incomplete data reporting. No beneficial dissemination strategies were found except one that looks promising, the use of peer educators in the worksite, which led to a short-term increase in fruit and vegetable intake. CONCLUSIONS AND IMPLICATIONS: Overall, the quality of the evidence is not strong and is primarily descriptive rather than evaluative. No clear conclusions can be drawn from these data. Controlled studies are needed to evaluate dissemination strategies, and to compare dissemination and diffusion strategies with different messages and different target audiences. PMID- 15819992 TI - Windows .NET Network Distributed Basic Local Alignment Search Toolkit (W.ND BLAST). AB - BACKGROUND: BLAST is one of the most common and useful tools for Genetic Research. This paper describes a software application we have termed Windows .NET Distributed Basic Local Alignment Search Toolkit (W.ND-BLAST), which enhances the BLAST utility by improving usability, fault recovery, and scalability in a Windows desktop environment. Our goal was to develop an easy to use, fault tolerant, high-throughput BLAST solution that incorporates a comprehensive BLAST result viewer with curation and annotation functionality. RESULTS: W.ND-BLAST is a comprehensive Windows-based software toolkit that targets researchers, including those with minimal computer skills, and provides the ability increase the performance of BLAST by distributing BLAST queries to any number of Windows based machines across local area networks (LAN). W.ND-BLAST provides intuitive Graphic User Interfaces (GUI) for BLAST database creation, BLAST execution, BLAST output evaluation and BLAST result exportation. This software also provides several layers of fault tolerance and fault recovery to prevent loss of data if nodes or master machines fail. This paper lays out the functionality of W.ND BLAST. W.ND-BLAST displays close to 100% performance efficiency when distributing tasks to 12 remote computers of the same performance class. A high throughput BLAST job which took 662.68 minutes (11 hours) on one average machine was completed in 44.97 minutes when distributed to 17 nodes, which included lower performance class machines. Finally, there is a comprehensive high-throughput BLAST Output Viewer (BOV) and Annotation Engine components, which provides comprehensive exportation of BLAST hits to text files, annotated fasta files, tables, or association files. CONCLUSION: W.ND-BLAST provides an interactive tool that allows scientists to easily utilizing their available computing resources for high throughput and comprehensive sequence analyses. The install package for W.ND-BLAST is freely downloadable from http://liru.ars.usda.gov/mainbioinformatics.html. With registration the software is free, installation, networking, and usage instructions are provided as well as a support forum. PMID- 15819994 TI - Research: Is resection of tumours involving the pelvic ring justified? : A review of 49 consecutive cases. AB - INTRODUCTION: Pelvic surgery is challenging and impacts significantly on limb and visceral function, thus, raising the question "is heroic surgery justifiable". This study assessed the functional, oncologic and surgical outcomes following pelvis tumour resections. METHODS: Between 1996-2003, 49 patients (mean age 43 years) underwent pelvic tumour resections- 38 primary malignant tumours, 5 secondary tumours and 6 benign tumours. Bone tumours comprised 5 osteosarcomas, 5 Ewings sarcomas, and 12 chondrosarcomas. Of the soft tumours, 9 were of neural origin. Tumours involved the ilium, acetabulum, pubic bones, sacrum or a combination of these. Functional assessment was performed and no patient had metastases at presentation. RESULTS: There were 41 limb sparing resections and 8 hindquarter amputations. Surgical margins were intralesional (1), marginal (13), wide (26), and radical (3). Of limb sparing surgery, prosthetic reconstructions were performed in 10 patients, biologic reconstructions in 6, a combination of these in 3 and no reconstruction in others. There was 1 intraoperative death, 7 local recurrences and 19 metastases. Death from disease occurred at a mean of 14.2 months with a mean followup of 27 (1-96) months. Amputation and periacetabular resections had worse functional outcomes. Emotional acceptance was surprisingly high. CONCLUSION: Pelvic resections are complex. Functional outcome is significantly affected by surgery. Disease control is similar to limb tumours. Emotional acceptance of surgery in survivors was surprisingly high. Major pelvic resection for malignancy appears justified. PMID- 15819993 TI - Hsp27 and axonal growth in adult sensory neurons in vitro. AB - BACKGROUND: Neurite growth can be elicited by growth factors and interactions with extracellular matrix molecules like laminin. Among the targets of the signalling pathways activated by these stimuli are cytoskeletal elements, such as actin, tubulin and neurofilaments. The cytoskeleton can also be modulated by other proteins, such as the small heat shock protein Hsp27. Hsp27 interacts with actin and tubulin in non-neuronal cells and while it has been suggested to play a role in the response of some neurons to injury, there have been no direct studies of its contribution to axonal regeneration. RESULTS: We have investigated neurite initiation and process extension using cultures of adult dorsal root ganglion (DRG) sensory neurons and a laminin stimulation paradigm. Employing confocal microscopy and biochemical analyses we have examined localization of Hsp27 at early and later stages of neurite growth. Our results show that Hsp27 is colocalized with actin and tubulin in lamellopodia, filopodia, focal contacts and mature neurites and growth cones. Disruption of the actin cytoskeleton with cytochalasin D results in aberrant neurite initiation and extension, effects which may be attributable to alterations in actin polymerization states. Inhibition of Hsp27 phosphorylation in our cultures results in an atypical growth pattern that may be attributable to an effect of pHsp27 on the stability of the actin cytoskeleton. CONCLUSION: We observed colocalization of the phosphorylated and non-phosphorylated forms of Hsp27 with actin and tubulin in both very early and later stages of neurite growth from cultured adult DRG neurons. The colocalization of Hsp27 and pHsp27 with actin in lamellopodia and focal contacts at early stages of neurite growth, and in processes, branch points and growth cones at later stages, suggests that Hsp27 may play a role in neuritogenesis and subsequent neurite extension, and potentially in the patterning of this growth. Hsp27 has been reported to play a key role in modulating actin cytoskeletal dynamics as an actin-capping protein in non-neuronal cells. Our results suggest that this may also be the case in neurons and support a role for Hsp27 in neurite outgrowth via its phosphorylation state-dependent interactions with actin. PMID- 15819995 TI - Absence of association between angiotensin converting enzyme polymorphism and development of adult respiratory distress syndrome in patients with severe acute respiratory syndrome: a case control study. AB - BACKGROUND: It has been postulated that genetic predisposition may influence the susceptibility to SARS-coronavirus infection and disease outcomes. A recent study has suggested that the deletion allele (D allele) of the angiotensin converting enzyme (ACE) gene is associated with hypoxemia in SARS patients. Moreover, the ACE D allele has been shown to be more prevalent in patients suffering from adult respiratory distress syndrome (ARDS) in a previous study. Thus, we have investigated the association between ACE insertion/deletion (I/D) polymorphism and the progression to ARDS or requirement of intensive care in SARS patients. METHOD: One hundred and forty genetically unrelated Chinese SARS patients and 326 healthy volunteers were recruited. The ACE I/D genotypes were determined by polymerase chain reaction and agarose gel electrophoresis. RESULTS: There is no significant difference in the genotypic distributions and the allelic frequencies of the ACE I/D polymorphism between the SARS patients and the healthy control subjects. Moreover, there is also no evidence that ACE I/D polymorphism is associated with the progression to ARDS or the requirement of intensive care in the SARS patients. In multivariate logistic analysis, age is the only factor associated with the development of ARDS while age and male sex are independent factors associated with the requirement of intensive care. CONCLUSION: The ACE I/D polymorphism is not directly related to increased susceptibility to SARS coronavirus infection and is not associated with poor outcomes after SARS coronavirus infection. PMID- 15819996 TI - Nuclear localization of Annexin A7 during murine brain development. AB - BACKGROUND: Annexin A7 is a member of the annexin protein family, which is characterized by its ability to interact with phospholipids in the presence of Ca2+-ions and which is thought to function in Ca2+-homeostasis. Results from mutant mice showed altered Ca2+-wave propagation in astrocytes. As the appearance and distribution of Annexin A7 during brain development has not been investigated so far, we focused on the distribution of Annexin A7 protein during mouse embryogenesis in the developing central nervous system and in the adult mouse brain. RESULTS: Annexin A7 is expressed in cells of the developing brain where a change in its subcellular localization from cytoplasm to nucleus was observed. In the adult CNS, the subcellular distribution of Annexin A7 depends on the cell type. By immunohistochemistry analysis Annexin A7 was detected in the cytosol of undifferentiated cells at embryonic days E5-E8. At E11-E15 the protein is still present in the cytosol of cells predominantly located in the ventricular germinative zone surrounding the lateral ventricle. Later on, at embryonic day E16, Annexin A7 in cells of the intermediate and marginal zone of the neopallium translocates to the nucleus. Neuronal cells of all areas in the adult brain present Annexin A7 in the nucleus, whereas glial fibrillary acidic protein (GFAP) positive astrocytes exhibit both, a cytoplasmic and nuclear staining. The presence of nuclear Annexin A7 was confirmed by extraction of the nucleoplasm from isolated nuclei obtained from neuronal and astroglial cell lines. CONCLUSION: We have demonstrated a translocation of Annexin A7 to nuclei of cells in early murine brain development and the presence of Annexin A7 in nuclei of neuronal cells in the adult animal. The role of Annexin A7 in nuclei of differentiating and mature neuronal cells remains elusive. PMID- 15819997 TI - Ethics and moral philosophy in the initiation of IVF, preimplantation diagnosis and stem cells. AB - Details of the work leading to the introduction of human IVF, animal and human stem cells, and the preimplantation diagnosis of inherited characteristics in blastocysts are outlined briefly in this paper. The progress of these studies is related to ethical issues emerging during these years. The current status of these studies is outlined, together with a brief moral philosophy as practised by the original investigators. PMID- 15819998 TI - Ethics, law and moral philosophy of reproductive biomedicine. Papers from an international conference at the Royal Society. September 30 - October 1, 2004. London, United Kingdom. PMID- 15819999 TI - Unusual families. AB - The introduction of assisted reproduction has led to unusual forms of procreation. This article describes the social consequences of lesbian motherhood and of families headed by single heterosexual mothers. PMID- 15820000 TI - Reproductive liberty, disease and disability. AB - There are many arguments from many sides, which purport to give reasons for limiting access to reproductive technologies and to gene-based reproductive procedures. There is one reason to reject them all, and that is that they do not point to dangers or harms of sufficient seriousness, probability or proximity to justify the limitation on human freedom that they require. This paper shows why this is so and argues that reproductive liberty must also include the liberty to select against disability both for moral and for libertarian reasons. PMID- 15820001 TI - The moral status of the human embryo: implications for IVF. AB - The Human Fertilisation and Embryology Act (1990) accords a special status to human embryos generated in vitro and this has resulted in strict limitations on the nature and extent of embryo research and fertility treatment that can be carried out in the UK. This special status derives from the embryo's recognized potential to become a human being. What do we understand by the concept of "human being", how does this differ from "conscious being" or "person" and how does this affect our attitude towards "microscopic clusters of cells" that may progress by an uncertain but inexorable process from the Petri dish to the crib? PMID- 15820002 TI - Discussion (day 1 session 1): Assisted conception and moral philosophy. PMID- 15820003 TI - Designing babies: what the future holds. AB - Advances in reproductive technology have opened new opportunities to avoid inherited diseases in offspring. The preimplantation genetic diagnosis (PGD) of human embryos permits those embryos carrying gene disorders or a non-diploid chromosome constitution to be identified. Numerous disease genes including those with a late onset have been identified and the conditions averted in children. Risks of abortion have been reduced, and the incidence of live births raised after PGD. It is also possible to select embryos with human leukocyte antigens (HLA) identical to those of a sick elder sibling, and then use stem cells from cord blood at birth to supply the necessary therapy. This form of treatment has alleviated the inherited disease in many recipients. The outlook and ethics of this approach to the alleviation of human disorders are discussed. PMID- 15820004 TI - Germinal choice technology and the human future. AB - This paper examines the likely impacts of emerging technologies that will give prospective parents the potential to directly influence the genetics of their offspring. My primary focus is on advanced prenatal genetic diagnosis (PGD) for both disease and non-disease traits, since this is likely to emerge before such possibilities as direct germline engineering. I place these technologies within the larger context of today's revolution in the life sciences and consider the progress likely to occur in this realm in the next few generations. I take a common sense look at the types of screening choices people are likely to make once these possibilities become possible, their broad consequences for human society, and the advantages and disadvantages of plausible regulatory paths in this realm. I also reflect upon today's debate about cloning and other such issues in the life sciences, looking at the driving forces behind these discussions and the tensions likely to develop in the next few decades. PMID- 15820005 TI - New breeds of humans: the moral obligation to enhance. AB - This paper argues that we have a moral obligation to enhance human beings. It is argued that if one is committed to the moral obligation to treat and prevent disease, one is also committed to genetic and other enhancement in so far as this promotes human well-being. It is argued that this is not eugenic but expresses our fundamental human nature: to make rational decisions and to try to improve ourselves. To be human is to strive to be better. PMID- 15820006 TI - Discussion (day 1 session 2): Designer babies. PMID- 15820007 TI - Reproductive cloning: past, present and future. AB - This brief outline in reproductive cloning describes the background to these studies and then discusses successive aspects of the subject. These include abnormalities in cloned animals, therapeutic cloning and the ethics of this subject. A reference to further reading is provided. PMID- 15820008 TI - The ethics of human reproductive cloning. AB - This article addresses the question of whether human reproductive cloning could be ethically justifiable in at least some cases involving infertile couples who would choose cloning as a way to have a genetically related child. At present, the risk of congenital anomalies constitutes a compelling argument against human reproductive cloning. The article explores whether reproductive cloning could be ethically justifiable if, at some future time, cloning becomes possible without an elevated risk of anomalies. It is argued that freedom to use cloning is a form of procreative freedom and, as such, deserves respect. All of the objections that have been raised against human reproductive cloning fall under three main categories: those that appeal to the interests of the child, those based on consequences for society, and those arising from teleological views. Objections that appeal to the child's interests are, in turn, of two main kinds: consequentialist and deontological. All of these types of objections are examined, and it is found that each involves serious problems that prevent it from being a reasonable objection in the context of the infertility cases considered. It is concluded that human reproductive cloning would be ethically justifiable in at least some cases involving infertile couples, provided that it could be performed without an elevated risk of anomalies. PMID- 15820009 TI - Human cloning and human dignity. AB - Judging from the official documents dealing with the moral and legal aspects of human reproductive cloning there seems to be a nearly worldwide consensus that reproductive cloning is incompatible with human dignity. The certainty of this judgement is, however, not matched by corresponding arguments. Is the incompatibility of reproductive with human dignity an ultimate moral intuition closed to further argument? The paper considers several ways by which the intuition might be connected with more familiar applications of the concept of human dignity, and argues that there is no such connection. It concludes that the central objections to human reproductive cloning are not objections relating to dignity but objections relating to risk, especially the risks imposed on children born in the course of testing the method's safety. PMID- 15820010 TI - Discussion (day 1 session 3): Moral philosophy of human reproductive cloning. PMID- 15820011 TI - Developments in human embryonic stem cells. AB - My task today is to describe the properties of stem cells speaking from the perspective of an investigator familiar with mammalian embryos and also working on human embryonic stem cells. My laboratory is based in the Department of Surgery and the Cambridge Stem Cell Institute, University of Cambridge, where you can find more information about ourselves and our colleagues (www.stemcells.cam.ac.uk). PMID- 15820012 TI - Embryonic stem cell research and the moral status of human embryos. AB - The paper analyses whether the old problem concerning the moral status or value of human embryos is implicated in the moral evaluation of embryonic stem cell research. It briefly outlines the moral status debate and then proceeds to show that all current attempts to bypass the problem fail. PMID- 15820013 TI - Stem cell research, personhood and sentience. AB - In this paper the permissibility of stem cell research on early human embryos is defended. It is argued that, in order to have moral status, an individual must have an interest in its own wellbeing. Sentience is a prerequisite for having an interest in avoiding pain, and personhood is a prerequisite for having an interest in the continuation of one's own existence. Early human embryos are not sentient and therefore they are not recipients of direct moral consideration. Early human embryos do not satisfy the requirements for personhood, but there are arguments to the effect that they should be treated as persons nonetheless. These are the arguments from potentiality, symbolic value and the principle of human dignity. These arguments are challenged in this paper and it is claimed that they offer us no good reason to believe that early human embryos should be treated as persons. PMID- 15820014 TI - Discussion (day 2 session 1): Stem cell outlook. PMID- 15820015 TI - Molecular biology of preimplantation embryos: primer for philosophical discussions. AB - This article is based on a presentation at the First International Conference on Ethics, Science and Moral Philosophy of Assisted Human Reproduction. The goal is to provide scientific background for the discussion of philosophic issues. Recent advances in the systematic molecular analysis of preimplantation embryos are summarized, including the molecular identification of nearly all genes involved in preimplantation development and their detailed expression patterns. Notwithstanding a quantum leap in molecular understanding of preimplantation embryos, molecular evidence seems to provide no decisive definition of a threshold for the beginning of human life during preimplantation development. PMID- 15820016 TI - The problematic in-vitro embryo in the age of epigenetics. AB - During use of many assisted reproductive technologies, the embryo spends time in vitro. The immediate and long-term epigenetic impacts of this exposure to an in vitro environment are discussed in the context of the health of the offspring. Three exemplary types of possible epigenetic impact are discussed: embryonic cell numbers, mitochondria, and genomic imprints. There is evidence that all of these can be affected in the short term and that these short-term impacts can have heritable consequences across developmental cell generations into maturity. There is also evidence of association between the observed impact and pathology, but as yet no unequivocal evidence of causality for humans and mice. The problematic in vitro embryo is considered paradigmatic for a central question facing biology: how does the environment interact epigenetically with the genome to produce variable phenotypic outcomes? PMID- 15820017 TI - Ethics and the future of preimplantation genetic diagnosis. AB - The future growth of preimplantation genetic diagnosis (PGD) will depend on refinements in genetic knowledge and genetic analysis of blastomeres. Equally important, however, is an acceptance of the ethical legitimacy of parents using technologies to select genetic traits of offspring. Objections based on embryo status, the giftedness of reproduction, eugenics, and protecting the child's welfare are not convincing grounds to oppose most uses of PGD. Whether PGD should be accepted for new medical or non-medical uses should depend upon a careful assessment of the proposed use's importance to the person or couple requesting it, and the harmful effects, if any, which it might cause. Such an approach leads to the conclusion that most new medical uses of PGD and some non-medical uses should be permitted. PMID- 15820018 TI - Gay science: assisted reproductive technologies and the sexual orientation of children. AB - There are no technologies at the present time that would allow parents to select the sexual orientation of their children. But what if there were? Some commentators believe that parents should be able to use those techniques so long as they are effective and safe. Others believe that these techniques are unethical because of the dangers they pose to homosexual men and women in general. Both sides point to motives and consequences when trying to analyse the ethics of this question. These arguments are reviewed, and it is concluded that opponents of these technologies have not shown good reason why the law or policy should override parental choice in this matter. In general, therefore, if technologies become available to choose the sexual orientation of children, parents should be allowed to use them, provided they are safe and disrupt no interest of the child. This use will, at the very least, protect homosexual children from parents who do not want them, but it will also allow parents who want homosexual children to make that choice as well. PMID- 15820019 TI - Discussion (day 2 session 2): Modern genetics and the human embryo in vitro. PMID- 15820020 TI - Scientific aspects of preconception gender selection. AB - Although numerous methods have been promoted as having an influence on the gender of offspring, most lack credible scientific evidence of effectiveness. Preconception gender selection has an important application in reducing the risk of having children with X-linked disease. A preconception, flow cytometric sperm sorting method of gender selection (MicroSort) is based upon the detection of differential fluorescence emitted by fluorescently stained X and Y chromosome bearing spermatozoa. Ongoing clinical trial results illustrate the safety and efficacy of the method. Fluorescence in-situ hybridization (FISH) analysis of specimens pre- and post-sort show the 50:50 X:Y ratio in unsorted spermatozoa can be shifted to 90% X or 75% Y after sorting. Embryo gender and fetal/baby gender are consistent with post-sort FISH results. Intrauterine insemination (IUI) and IVF/ICSI (intracytoplasmic sperm injection) pregnancy rates are consistent with those reported in the assisted reproduction literature. Clinical loss rates are similar to those in the general population. The observed major congenital abnormality rate of 2.05% for babies born after using this sperm sorting method coincides with that occurring spontaneously. Current results indicate that the method is a safe and effective process and is a valuable tool for families wishing to balance the gender distribution of their children or to reduce the likelihood of having children with X-linked disease. PMID- 15820021 TI - Preconception gender selection: a threat to the natural sex ratio? AB - This brief paper summarizes a series of postal investigations on the acceptance of selection for X or Y spermatozoa. These were conducted mainly in Germany but also in the UK, the Netherlands and the US. Selected families were approached with a series of questions about their wish to use sperm selection, and their choice of boys or girls. In general, large majorities opposed this approach for family balancing or sex selection on the basis of cost and inconvenience of the treatment. The view was expressed that its use in medicine was more acceptable. Care was taken to compose a series of open questions to particular sections of the population, e.g.men versus women, differing ages, and families with imbalanced sex ratios. More favourable responses arose, for example in pregnant women. Overall, all four nations had large majorities against the use of sex selection. PMID- 15820022 TI - Choosing children: intergenerational justice? AB - In this discussion, we argue that the concept of intergenerational justice, usually used in environmental matters, is applicable to reproductive decisions also. Additionally, we propose that this permits certain reproductive choices to be made prior to conception or during the pregnancy, and that these choices should not be confined to clinical concerns. In particular, we argue that consideration of the interests of future children should be viewed from the perspective of objective well-being. That being the case, decisions about the sex of future offspring can, in terms of intergenerational justice, be legitimate. We do not argue that every reproductive choice is legitimate; for example it would not be legitimate deliberately to choose characteristics that prevent future children from potentially successful participation in social life. PMID- 15820023 TI - Discussion (day 2 session 3): Ethics of choosing the sex of our offspring. PMID- 15820024 TI - The morality of screening for disability. AB - Many people object to preimplantation or prenatal screening for disability on the grounds that it is discriminatory, has pernicious effects on the lives of existing disabled people, expresses a hurtful view of disabled people, and reduces human diversity. I argue that if these objections are held to be strong enough to show that screening is wrong, they must also imply the permissibility of causing oneself to have a disabled rather than a non-disabled child. Indeed, those who object to screening on these grounds and also claim that it is not worse to be disabled than not to be, seem to be committed to accepting the permissibility of deliberately causing disabling prenatal injury, even for frivolous reasons. If we cannot accept these implications, we cannot accept that the objections to screening show that it is wrong. PMID- 15820025 TI - Eugenics: some lessons from the past. AB - Eugenics was first debated by the ancient Greeks, particularly Plato and Aristotle, developed in the nineteenth century by Francis Galton and Charles Darwin, and then abused in the twentieth century by right-wing politicians. With the new methods of assisted conception combined with the use of genetic markers, all the old problems of eugenics have resurfaced. Gender selection, embryo selection, preimplantation genetic diagnosis of common disease, and gene replacement techniques (somatic cells) have added greatly to the power of the modern eugenicist. How are these procedures to be monitored and regulated? What is the role of the State compared with individual families for the implementation of the new methodologies? Some of these issues will be discussed. PMID- 15820026 TI - Godless morality. AB - Because of the transcendental claims they make, debating human ethics with religious figures can be frustrating, but two things might help. First of all, we should refuse to be bullied by them and should remain firm in asserting the fact that though our ethic may differ from theirs, it is an ethic, not the lack of one. Secondly, we should recognize that religions evolve and adapt to new understandings of human nature, but they have to make the changes within the scope of their own self-understanding. Knowing that the best of them do this, we should, where possible, co-operate with them in negotiating the rapids of social and ethical change. PMID- 15820027 TI - Reproductive freedom and the State. AB - This brief paper describes the experiences and attitudes of a UK Member of Parliament to the politics of reproductive biomedicine. It provides insights into how problems in this field strike politicians and governments, and their interactions with the press and the public. It takes examples of four topics which help to decide the attitudes of legislators in such complex ethical matters. It also reveals how they can change their opinions as they became familiar with the ethical issues raised under various situations. PMID- 15820028 TI - Discussion (day 2 session 4): Lessons on ART from philosophy, religion and politics. PMID- 15820031 TI - The dilemma of recurrent ectopic pregnancy after tubal surgery and in-vitro fertilization. What to do? AB - This commentary was written to stimulate a debate on the role of tubal surgery for ectopic pregnancy in women undergoing IVF. PMID- 15820032 TI - Would 'all-inclusive' compensation attract more gamete donors to balance their loss of anonymity? AB - The UK Government has determined that children born from egg donation have the same right to know their biological parent as adopted children on reaching the age of 18. There is concern as to the effect of loss of anonymity on egg donor recruitment, since a wait of 1 or 2 years is not unusual even under present circumstances. Some fertility programmes have introduced egg-sharing (involving 'subfertile donors') to circumvent the delays. However, with this method both the donor and the recipient may have reduced overall success. The shortage of sperm donors has led the Human Fertilisation and Embryology Authority (HFEA) to consider increasing the allowable expenses payment and a consultation exercise is underway to review payments and other issues for sperm, egg and embryo donors. Fundamental new initiatives regarding gamete donation need to be implemented to avoid further restrictions on an already unsatisfactory service. An 'all inclusive' financial allowance is proposed for donors to promote donor recruitment and shorten the waiting time to treatment predicted to increase with the loss of anonymity. PMID- 15820033 TI - Commentary on Craft and Thornhill: new ethical strategies to recruit gamete donors. AB - The main problem for the procedure of gamete donation at present is that the abolition of the donor anonymity rule has destroyed coherence among the guidelines. To restore coherence, a new unified picture of the role of the gamete donor should be constructed. It is doubtful that payment will increase the number of candidates in a system with identifiable donors. Nevertheless, if non financial rewards of donors do not suffice to motivate candidates, an 'all inclusive' financial allowance analogous to volunteers in medical trials could be considered to prevent a complete collapse of the system. PMID- 15820034 TI - Risks and ethics of gene transfer. PMID- 15820035 TI - Assisted reproductive practice: religious perspectives. AB - It is important to those who practise reproductive techniques to learn about different religious perspectives related to reproductive health problems. Religious groups are active in influencing the public regarding bioethical positions, and this is particularly evident with issues concerning procreation, abortion and infertility therapy. The Jewish attitude towards procreation is derived from the first commandment of God to Adam to 'Be fruitful and multiply'. Judaism allows the practice of all techniques of assisted reproduction when the oocyte and spermatozoon originate from the wife and husband respectively. The attitude toward reproductive practice varies among Christian groups. While assisted reproduction is not accepted by the Vatican, it may be practised by Protestant, Anglican and other denominations. According to traditional Christian views, beginning at conception, the embryo has moral status as a human being, and thus most assisted reproductive technologies are forbidden. According to Islam, the procedures of IVF and embryo transfer are acceptable, although they can be performed only for husband and wife. Developments in science and technology and corresponding clinical applications raise new religious questions, often without clear answers. The role of theology in bioethics is integral to clarify perceived attitudes toward these developments for different religious communities. This paper presents the attitude of monotheistic religions to therapeutic procedures, such as IVF-embryo transfer, spermatozoa, oocytes, embryo donation, cryopreservation of genetic material, surrogacy, posthumous reproduction, gender preselection, reproductive and therapeutic cloning. PMID- 15820036 TI - Ethical consideration of intentioned preimplantation genetic diagnosis to enable future tissue transplantation. AB - Recently, a case was presented where a couple with one normal child requested preimplantation genetic diagnosis (PGD) with human leukocyte antigen (HLA) typing in order to conceive children who could serve as mutual donors for future transplantations if needed. While PGD for medical reasons is well defined and accepted, application for non-medical conditions is less obvious and still in debate. The present case calls to further extend the indications for PGD and raises a moral question as to whether to allow such intentioned treatment. Although it seems that all siblings might benefit from the procedure, when weighing the risks benefits and the ethical issues included in performing intentioned PGD, it seems that the procedure harbours risks and ethical issues that outweigh the benefit it offers. Therefore, it is time to constrain the slippery slope and state that intentioned PGD for conceiving children who might serve as siblings for mutual tissue donations should be declined. PMID- 15820037 TI - Will removal of anonymity influence the recruitment of egg donors? A survey of past donors and recipients. AB - The UK Government has determined that children born from egg donation have the same right to know their biological parent as adopted children on reaching the age of 18. There is concern as to the effect of loss of anonymity on egg donor recruitment, since a wait of 1 or 2 years is not unusual for couples awaiting treatment. Some fertility programmes have introduced egg-sharing to circumvent these delays. However, egg-sharing may involve a sub-fertile donor and thus is likely to be less successful overall than egg donation. Recently, the Human Fertilisation and Embryology Authority announced a consultation exercise to consider whether egg donors should receive increased payment or benefits in kind for their donation. To investigate whether or not removal of anonymity would have influenced or would influence their future participation, past egg donors and recipients have been surveyed. Questionnaires were sent to 867 former egg donors (n=504) and recipients (n=363), who were anonymous to each other. Response rates were similar between donors (32.7%) and recipients (39.1%). Of donors, 36.4% would not have participated had donor anonymity been waived, but 69.1% would donate anonymously again. Of recipients, 53.5% would not have proceeded had donor anonymity been waived, whereas 96.5% would receive anonymously donated eggs again. The results indicate that removal of anonymity for egg donors is likely to lead to a further restriction of an already unsatisfactory service to patients requiring donated eggs unless fundamental new initiatives are implemented. PMID- 15820038 TI - Use of antagonists in ovarian stimulation protocols. AB - Gonadotrophin-releasing hormone (GnRH) antagonists have been introduced in IVF to prevent premature LH surge. They bind competitively to pituitary receptors and prevent endogenous GnRH from exerting any stimulus on pituitary cells, avoiding the initial 'flare-up' effect and decreasing gonadotrophin secretion within a few hours. Pituitary reserve and gonadotrophin synthesis are not affected; therefore, the recovery of pituitary function is rapid. Two different regimes have been described. The multiple-dose protocol involves the administration of 0.25 mg cetrorelix (or ganirelix) daily from day 6-7 of stimulation, or when the leading follicle is 14-15 mm, until human chorionic gonadotrophin (HCG) administration. The single-dose protocol involves the single administration of 3 mg cetrorelix on day 7-8 of stimulation. Both antagonists with either regimen seem to be equally effective in the prevention of the LH surge. Compared with a long luteal agonist protocol, the treatment is shorter and requires a smaller amount of gonadotrophins. Pregnancy rate seems to be lower, but a decrease in the incidence of severe ovarian hyperstimulation syndrome (OHSS) is reported by several studies. A promising aspect of antagonists may be the possibility of making treatment less aggressive. Finally, in antagonist cycles, ovulation triggering is possible by GnRH agonists, avoiding the deleterious effect of HCG and thus preventing OHSS. PMID- 15820039 TI - Prolactin receptor mRNA expression in oocytes and preimplantation mouse embryos. AB - Prolactin was first identified as an anterior pituitary lobe hormone, responsible for the regulation of mammary gland growth and development. Prolactin receptors have been localized in a number of peripheral tissues, including tissues involved in reproduction. Studies with knockout animals have shown that prolactin receptor deficient mice present reproductive defects, whereas prolactin promotes the developmental potential of preimplantation mouse and rat embryos in vitro. To better understand the role of prolactin in the process of reproduction and early embryo development in mice, the expression of the four transcript variants of prolactin receptor was examined in the first stages of mouse embryo development. Prolactin long receptor mRNA was expressed in all stages examined, that is in cumulus cells, oocytes, zygotes, 2-cell embryos, 4-cell embryos, morulae and blastocysts. Prolactin receptor type S1 mRNA was observed only in cumulus cells, while S2 mRNA was present in cumulus cells, oocytes, zygotes and 2-cell embryos. S3 mRNA was expressed only in cumulus cells and oocytes. These results indicate that different isoforms of prolactin receptors may be present in the various stages of mouse preimplantation embryo and may play an important role in the control of its growth and development. PMID- 15820040 TI - Achievement of pregnancy three times in the same patient during luteal GnRH agonist administration. AB - Gonadotrophin-releasing hormone agonist (GnRHa) administration from the mid luteal phase onwards is considered the gold standard of ovarian stimulation for IVF treatment. It might, however, coincide with an implanting spontaneous pregnancy. Concerns have therefore been raised with regard to the evolution of the resulting pregnancies and long-term outcome of the children born. The current case report describes the achievement of three pregnancies in the same patient during luteal administration of GnRHa. One pregnancy ended in spontaneous abortion and the other two resulted in the delivery of two female infants. The children have so far been followed for 3.5 and 7 years. The physical examination of both children was unremarkable. However, the older child has recently been diagnosed with attention deficit hyperactivity disorder and dyslexia. PMID- 15820041 TI - Clean technique for cryoprotectant-free vitrification of human spermatozoa. AB - Human spermatozoa can be successfully cryopreserved without the use of cryoprotectants through vitrification at very high warming rates. This is achieved by plunging a small amount of frozen sperm suspension into a warming medium, or a large amount of sperm suspension into an agitated warming medium. The aim of the present study was to compare the motility of human spermatozoa cryopreserved using four different methodologies of cooling and warming: cryoloops, droplets, open-pulled straws and standard open straws. Evaluation of two parameters, motility and viability rate of spermatozoa, suggests that all four methods are suitable for use in assisted reproductive technology. However, only the use of open-pulled straws as well as standard open straws allows the isolation of spermatozoa from liquid nitrogen with low potential risk of microbial contamination during freezing and storage, and is thereby a clean method of vitrification. PMID- 15820043 TI - Culture methods influence haemopoietic and endothelial differentiaion of ES cells. PMID- 15820042 TI - Combinatorial peptide library binding of mammalian spermatozoa identifies a ligand (HIPRT) in the axin protein: putative identification of a sperm surface axin binding protein and intriguing developmental implications. AB - The identification of components in cell-cell interactions is an important research goal in reproductive and developmental biology. Such interactions are critical to gamete development, fertilization, implantation and basic development. Several proteins involved with sperm-oocyte interaction and other developmentally important phenomena have been identified. However, these are obviously only a subset of the molecular components involved in such complex cell cell interactions. One method that has been used to identify binding partners for particular molecular targets is the use of combinatorial libraries accessible on phage surfaces. For the most part, this technique has mainly been applied to screen specific target moieties. However, in some cases whole-cell screening has been attempted. This study describes the first report of screening intact, living mammalian gametes using a proprietary whole-cell combinatorial library binding and analysis protocol. Results from the first screening protocol of mouse spermatozoa strongly identified a putative sperm-binding ligand using proprietary bioinformatic analysis. This amino acid sequence (HIPRT) precisely corresponds with a previously characterized highly conserved protein-protein interaction site in the axin protein. This sequence is found within the binding site for a known sperm surface protein, glycogen synthase kinase-3. This result not only provides proof of the utility of this technique to identify cell surface ligands in mammalian gametes, but it also suggests a potential role for spermatozoa in facilitating developmental axis formation in mammalian embryos. PMID- 15820044 TI - Negligible interchromosomal effect in embryos of Robertsonian translocation carriers. AB - It has been suggested that translocations, and perhaps other chromosome rearrangements, disturb meiotic disjunction of chromosome pairs not involved in the translocation, resulting in non-disjunction in those chromosomes (interchromosomal effect) and predisposition to trisomy offspring. Other reports have suggested an increased risk of mosaicism and chaotic embryos in translocation carriers. This study was designed to determine if such interchromosomal effects are producing significantly more chromosome abnormalities than those expected from unbalanced gametes. For that purpose, two groups of PGD patients were compared, Robertsonian translocation carriers (RBT) and carriers of X-linked diseases (XLI), of similar age. Both groups were analysed by FISH with similar DNA probes. The results indicate that overall, the higher rate of chromosome abnormalities in the RBT group was solely due to unbalanced gametes and not to an interchromosomal effect or higher incidence of mosaicism. If unbalanced and normal were combined, this proportion was 53% in XLI and 59% in RBT. However, when specific RBT translocations were studied, only a slight increase in embryos with aneuploidy for chromosome 22 was found for the t(13;14) translocation carriers, while a higher rate of post-zygotic abnormalities was observed in the more rare RBT. In conclusion, the overall rate of non-translocation related abnormalities was not increased in the RBT group compared with the control group, but a slight interchromosomal effect may exist, as some Robertsonian translocations may be more prone to produce mosaic and chaotic embryos. PMID- 15820045 TI - Paternal effects on cell division in the human preimplantation embryo. AB - Cell divisions in the human preimplantation embryo can be compromised by deficiencies in sperm nuclear genome or sperm-derived developmentally relevant cytoplasmic factors, oocyte activating substance and centriole. Sperm nuclear deficiencies are usually not detected before the 8-cell stage of embryo development, when a major expression of sperm-derived genes has begun. Sperm cytoplasmic deficiencies can be detected as early as the 1-cell zygote and then throughout the preimplantation development. The terms 'late paternal effect' and 'early paternal effect' have been suggested to denote these two pathological conditions. The late paternal effect is associated with an increased incidence of sperm DNA fragmentation. No association with sperm DNA damage has been found for the early paternal effect. The diagnosis of the late paternal effect is thus based on the examination of sperm DNA integrity, which should be performed in cases of repeated assisted reproduction failure even if morphologically normal embryos result from fertilization with the patient's spermatozoa. The only element leading to the diagnosis of the early paternal effect is poor zygote and embryo morphology and low cleavage speed. The absence of increased sperm DNA damage does not exclude the presence of this pathology. ICSI with testicular spermatozoa has recently been shown to be an efficient treatment for the late paternal effect. The use of oral antioxidant treatment in this indication has also given promising results. PMID- 15820046 TI - Clinical application of multiple displacement amplification in preimplantation genetic diagnosis. AB - Multiple displacement amplification (MDA) is a technique used in the amplification of very small amounts of DNA. MDA is reported to yield large quantities of high-quality DNA. The applicability of MDA to single cells was recently demonstrated as a potential technique for preimplantation genetic diagnosis (PGD). This paper shows the first clinical application of MDA in PGD. Two cycles of PGD were performed in two diseases, resulting in two pregnancies. All the diagnoses given on blastomeres were confirmed on the non-transferred whole embryos. The blastomere diagnosis was coupled with short tandem repeat (STR) analysis (16 loci) in all cycles. Allelic drop-out (ADO) assessment and amplification efficiency were evaluated on 40 single lymphocytes derived from parents of each disease. ADO and amplification failure were 10.3 and 2.2% for beta-thalassaemia and 17.9 and 2.2% for cystic fibrosis respectively. HLA matching for A, B and DR was performed successfully on single cell for the beta thalassaemia family using similar methods to genomic DNA. The PGD protocol used in all diseases consists of MDA amplification, followed by a standard polymerase chain reaction protocol. Although HLA matching was not applied to embryos, its feasibility was shown on single cell DNA amplified by MDA. Altogether, these data show the simplicity and reliability of performing PGD in combination with HLA matching and STR analysis using MDA. PMID- 15820047 TI - Preimplantation genetic diagnosis for aneuploidy screening in repeated implantation failure. AB - Chromosomal abnormalities are thought to be responsible for implantation failure, and among chromosomal abnormalities in normally developing embryos, aneuploidy is the most frequent. Genetic testing of preimplantation embryos for chromosomal aneuploidy allows selection of chromosomally normal embryos, and early detection of chromosomal aberration will increase the chance of conceiving. Preimplantation genetic diagnosis for aneuploidy screening (PGD-AS), performed by polar body or blastomere analysis, is used in infertile patients treated with assisted reproduction technologies, especially in those with a poor prognosis, e.g. repeated IVF failure, advanced maternal age, or recurrent spontaneous abortion. The aim of this paper is to clarify the impact of PGD-AS in repeated implantation failure. In this review, the data collected so far regarding PGD-AS in this patient group will be discussed in depth. PMID- 15820048 TI - Episodes of anxiety affect ICSI outcomes. PMID- 15820049 TI - Ovarian cancer. PMID- 15820051 TI - Pregnancy resulting from transfer of repeat vitrified blastocysts produced by in vitro matured oocytes in patient with polycystic ovary syndrome. AB - This report describes a live birth produced from repeat vitrification and thawing of blastocysts derived from in-vitro matured (IVM) oocytes in a woman with polycystic ovarian syndrome. Immature oocyte retrieval was performed on day 12 of her induced menstrual cycle. The patient was administered 10,000 IU of human chorionic gonadotrophin s. c. 36 h before immature oocyte retrieval. A total of 47 immature oocytes were collected. Following IVM of these immature oocytes, 76.6% (36/47) become mature (at metaphase II stage). Thirty oocytes (30/36, 86.1%) were normally fertilized following insemination by intracytoplasmic sperm injection. The fertilized zygotes (two-pronuclear stage) were co-cultured with cumulus cells in YS medium supplemented with 10% human follicular fluid. On day 5 after insemination, three blastocysts were transferred. Unfortunately, fresh embryo transfer did not result in pregnancy. The remaining 10 embryos developed to the expanded blastocyst stage. These remaining blastocysts were vitrified with electron microscope grids following artificial shrinkage. Three months later, three blastocysts were thawed due to a clinical error. Consequently, the embryos were revitrified. After a week, the three blastocysts were warmed again. Two of them developed to hatched blastocysts. Following transfer, a full-term pregnancy resulted in the delivery of healthy twins. PMID- 15820050 TI - Generation of a human embryonic stem cell line encoding the cystic fibrosis mutation deltaF508, using preimplantation genetic diagnosis. AB - Human embryonic stem (hES) cells are pluripotent cells isolated from early human embryos. They can be grown in vitro and made to differentiate into many different cell types. These properties have suggested that they may be useful in cell replacement therapy for many degenerative diseases. However, if hES cells could also be manufactured with mutations significant in human disease, they could provide a powerful in-vitro tool for modelling disease processes and progression in a number of different cell types, as well as providing an ideal system for studying in-vitro toxicity and efficacy of drugs and other therapeutic systems such as gene therapy. Embryos with such mutations are generated as part of routine genetic testing during preimplantation genetic diagnosis, providing the opportunity to generate cell lines with significant mutations. A human embryonic stem cell line homozygous for the most common mutation leading to cystic fibrosis in humans (delta F508) has been generated and characterized. This cell line has the same morphology and expresses proteins typical of other unaffected hES cell lines. This cell line represents an important in-vitro tool for understanding the pathophysiology of cystic fibrosis, and presents exciting opportunities to test the efficacy and toxicity of new therapies relevant to CF. PMID- 15820052 TI - Genomic microarrays provide insights into structural chromosomal rearrangements. PMID- 15820053 TI - Reduction of high order multiples in frozen embryo transfers. AB - The aim of this study was to determine if blastocyst frozen embryo transfers are able to reduce the potential for the rate of high order multiples (HOM) (>2 fetal sacs) without affecting the overall pregnancy and implantation potential. Group A included all frozen blastocyst transfers prior to 1 January 2002. Group B included all frozen blastocyst transfers between 1 January 2002 and 12 December 2003. There was no significant difference for survival for the two time periods (79 versus 80%). A significantly (P<0.05) lower number of embryos were transferred in group B (1.9) compared with group A (2.8). There was a significant (P<0.05) reduction of (HOM) from 31 to 3% for groups A and B respectively. Ongoing pregnancy rates for group A resulted in 52% of 25 embryo transfers and 41% of 75 embryo transfers in group B (not significant). Reducing the number of embryos transferred between groups A and B did not significantly impact implantation rates. It is concluded that blastocyst freezing is effective for overall survival, pregnancy and implantation while reducing the rate of high order multiples. PMID- 15820054 TI - Wider reproductive choices in assisted conception. PMID- 15820055 TI - Paraffin-embedded manipulated blastocysts: a tool to demonstrate stem cell plasticity? AB - One of the big question marks in current stem cell research is whether there is true plasticity of adult progenitor cells (APC) or if cell fusion is the principle source of the supposed plasticity. The generation of chimeras by injecting adult progenitor cells into blastocysts is not new. This paper describes an efficient embedding technique for murine blastocysts injected with human APC. This method could help in establishing a novel tool to analyse the process of plasticity, if it truly exists. If this is the case, this technology could be of great help to characterize surface markers of stem cells in great detail. On the other hand, fusion of cells could also be investigated. A system of embedding blastocysts was set up using paraffin for further analysis by means of light microscopy and immunohistochemistry. The embedding of the chimaeras consists of fixing them first with paraformaldehyde in phosphate-buffered saline (PFA/PBS), embedding them in gelatine, fixing the gelatine block with PFA/PBS and finally fixing the gelatine block in a Petri dish by embedding it in paraffin. Using this protocol, the morphology of the blastocysts is well preserved. PMID- 15820056 TI - Earliest detection of pregnancy. PMID- 15820057 TI - [Differential expression of STRBP8, a new candidate oncogene, in cancerization of human immortalized esophageal epithelial cells]. AB - BACKGROUND & OBJECTIVE: Recently, changes in composition, structure, and function of nuclear matrix proteins (NMPs) in generation and development of tumors evoked more and more attention. Separation and identification of tumor-related NMPs is a new way to search for tumor specific biomarkers, and to study tumor pathogenesis. This study was to analyze differential expression of STRBP8, one of esophageal carcinoma specific NMPs, in cancerization of immortalized human esophageal epithelial cells. METHODS: NMPs were extracted from immortalized human esophageal epithelial cell line SHEE and malignantly transformed esophageal carcinoma cell line SHEEC. Differential expression of STRBP8 was detected by two-dimensional electrophoresis (2-DE), matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS), and reverse transcription-polymerase chain reaction (RT-PCR). STRBP8 cDNA obtained by RT-PCR was linked to pGEM-T easy vector, and introduced into TOP10F' E.coli competent cells. Positive clones were sequenced and analyzed with BLAST. RESULTS: STRBP8 was only detected in SHEEC cells by 2-DE, MALDI-TOF-MS, and RT-PCR. The sequence of positive clones contained STRBP8 cDNA was identical to that in GenBank database. CONCLUSION: STRBP8, as a candidate oncogene, might relate to cancerization of esophageal epithelial cells, which might be a specific biomarker of esophageal carcinoma, and probe into the pathogenesis of esophageal carcinoma. PMID- 15820058 TI - [Antitumor effects of recombinant quail vascular endothelial growth factor receptor 2, as a vaccine, combined with cisplatin on LL/2 tumor model in mice]. AB - BACKGROUND & OBJECTIVE: Bio-chemotherapy is a new trend in cancer therapy. Antiangiogenic therapy represents a new strategy of tumor biotherapy. This study was designed to explore antitumor effect of recombinant quail vascular endothelial growth factor receptor 2 (qVEGFR) combined with cisplatin. METHODS: Lewis lung carcinoma model (LL/2) was established in C57BL/6 mice. Seven days after inoculation of tumor cells, mice were randomized into combination group, qVEGFR group, chemotherapy group, and normal saline (NS) group (10 mice/group), and received relevant treatments. Tumor growth, survival rate of mice, and side effects were observed. Anti-VEGFR-2 antibody-producing B cells (APBCs) was detected by enzyme-linked immunospot (ELISPOT), microvessel density (MVD) of tumor was detected by immunohistochemistry, and tumor cell apoptosis was also detected. RESULTS: Tumor volume of mice was obviously smaller in combination group than in NS group. Complete regression of tumor growth was observed in 3 of the 10 mice in combination group. Seventy days after inoculation of tumor cells, survival rate of mice was significantly higher in combination group than in qVEGFR, cisplatin, and NS groups (90% vs. 60%, 0%, and 0%, P < 0.05). APBCs counts were (156.8+/-19.3)/10(6) spleen cells in combination group, and (143.6+/ 18.6)/10(6) spleen cells in qVEGFR group. MVD was significantly lower in combination group than in cisplatin, and NS groups (11.4+/-1.3 vs. 33.4+/-4.5, and 45.5 +/- 4.5, P < 0.01), MVD in qVEGFR group was 16.4+/-1.6. Tumor cell apoptosis was significantly higher in combination group than in qVEGFR, cisplatin, and NS groups [(36.2+/-3.5)% vs. (15.4+/-2.4)%, (17.6+/-2.6)%, and (4.1+/-1.4)%, P < 0.05]. CONCLUSION: The combination therapy of qVEGFR and cisplatin has synergistic antitumor effect. PMID- 15820059 TI - [Antitumor effects of interferon-gamma-inducible protein 10 combined with gemcitabine]. AB - BACKGROUND & OBJECTIVE: Interferon-gamma-inducible protein 10 (IP-10), a member of alpha-chemoattractant family, is an inhibitor of tumor angiogenesis. Chemotherapy combined with antiangiogenic therapy has synergistic effects. This study was designed to explore antitumor effect of IP-10 combined with gemcitabine in tumor-bearing mice. METHODS: pBLAST-IP-10 was transferred into E.coli to prepare and purify plasmid IP-10. Hepatocarcinoma H22 model was established in BALB/c miceu Lewis lung cancer LL/2 model was established in C57BL/6 mice. The mice were divided into 4 groups, and treated with IP-10, gemcitabine, IP-10 plus gemcitabine, and normal saline (control), respectively. Protein level of IP-10 in serum was detected by ELISA. Survival rate of mice, tumor volume, and side effects of treatment were observed. Microvessel density (MVD) of tumor tissue was detected by immunohistochemistry. Apoptosis of tumor cells was detected by TUNEL method. RESULTS: After treatment of IP-10, IP-10 protein reached the peak level of (16.8+/-3.6) ng/ml at the 14th day, and remained a high level of (14.0+/-2.1) ng/ml at the 35th day. Compared with the controls, tumor volume of mice in the combination therapy group had significant regression, or disappeared. Nine weeks after inoculation of tumor cells, survival rate of mice was significantly higher in combination group than in IP-10, gemcitabine, and control groups (90% vs. 55%, 0%, and 0%, P < 0.005). No obvious side effects were observed. MVD of tumor tissues was significantly lower in combination group than in IP-10, gemcitabine, and control groups (15.8+/-2.4 vs. 45.6+/-2.0, 50.2+/-3.5, and 51.3+/-3.0, P < 0.01). At the 35th day after inoculation of tumor cells, apoptosis index was significantly higher in combination group than in IP-10, gemcitabine, and control groups (85.5+/-10.2 vs. 21.4+/-5.5, 8.4+/-2.0, 4.2+/-0.7, P < 0.01). CONCLUSION: Therapy of IP-10 combined with gemcitabine has significantly synergistic antitumor effect compared with IP-10 or gemcitabine alone. PMID- 15820060 TI - [Enhancement effect of nucleotide excision repair gene xeroderma pigmentosun group a antisense RNA on sensitivity of human lung adenocarcinoma cell line A549 to cisplatin]. AB - BACKGROUND & OBJECTIVE: Enhanced nucleotide excision repair (NER) capacity is an important mechanism of drug-resistance of tumor cells. Xeroderma pigmentosun group A (XPA) gene plays a key role in early stage of NER pathway. This study was to explore correlation between down-regulation of XPA gene induced by antisense RNA transfection and sensitivity of human lung adenocarcinoma cell line A549 to cisplatin. METHODS: A549 cells were stably transfected with XPA antisense RNA. Positive cell clones were selected by limiting dilution. Northern blot and Western blot were applied to evaluate mRNA and protein levels of XPA in positive cell clones. Sensitivity of A549 cells to cisplatin was evaluated by MTT assay. Host cell reactivation (HCR) assay was used to assess NER capacity of cisplatin damaged A549 cells. RESULTS: Six positive cell clones, AS1-AS6, were obtained, mRNA and protein levels of XPA were significantly decreased in AS3-AS6 cells. In dose-dependent experiment, the 50% inhibitory concentrations (IC(50)) of cisplatin to parental A549 cells and AS1-AS6 cells were 8.1, 7.6, 4.7, 3.2, 1.9, 2.8, and 4.1 mug/ml, respectively. Sensitivities of AS3-AS6 cells were significantly higher than that of parental A549 cells (F = 9.75, 9.14, 7.39, 8.91u P = 0.005, 0.006, 0.012, 0.006). In addition, mRNA level of XPA was positively correlated with IC(50) of cisplatin (r = 0.927, P = 0.003). Time effect experiment also showed increases of sensitivity to cisplatin in AS3-AS6 cells. HCR assay showed that NER capacities of AS3-AS6 cells were reduced. When treated with 40, 200, and 1 000 ng/ml of cisplatin, mRNA levels of XPA were positively correlated with cellular NER capacities (r = 0.854, 0.696, 0.858u P = 0.014, 0.082, 0.013). CONCLUSION: The targeted inhibition of XPA by antisense strategy can significantly decrease mRNA level of XPA, reduce cellular NER capacity, and sensitize lung cancer cells to cisplatin. PMID- 15820061 TI - [Impact of hypoxia on taxol-induced apoptosis in human ovarian cancer cell line A2780 and its mechanism]. AB - BACKGROUND & OBJECTIVE: Hypoxia, a feature and important living microenvironment of solid tumors, might be related to drug-resistance of tumors. This study was to establish a hypoxic model of ovarian cancer cell line A2780, and to investigate impacts of hypoxia and hypoxia inducible factor-1alpha (HIF-1alpha) on Taxol induced apoptosis in A2780 cells. METHODS: Cobalt chloride (CoCl(2)), a chemical hypoxia inducer, was added in A2780 cells to develop a hypoxic model. A2780 cells were divided into 4 groups: group A (control), group B (normoxia plus Taxol), group C (hypoxia plus Taxol), and group D (hypoxia, Decoy plus Taxol). Decoy method was used to block the function of HIF-1alpha. Protein and mRNA levels of HIF-1alpha were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL), and flow cytometry (FCM). RESULTS: CoCl(2) evidently increased protein level of HIF-1alpha, but had no effect on mRNA level of HIF 1alpha. Decoy didn't affect its expression. TUNEL showed that apoptotic index (AI) was significantly higher in group B,and group D than in group C after treatment of Taxol [(41.1+/-25.6)%,and (35.2+/-21.7)% vs. (24.1+/-15.2)%, P < 0.05]. The apoptosis rates detected by FCM displayed the same tendency as TUNEL results did. CONCLUSIONS: Hypoxia could weaken the effect of Taxol on inducing apoptosis of A2780 cells. HIF-1alpha might confer to resistance of cell apoptosis induced by Taxol. PMID- 15820062 TI - [Regulatory effect of STAT5 decoy oligonucleotides on trans-activation of bcl-x gene promoter in K562 cells]. AB - BACKGROUND & OBJECTIVE: Constitutive activation of signal transducers and activators of transcription 5 (STAT5) plays an important role in malignant transformation of chronic myeloid leukemia (CML) cells. This study was to explore regulatory effect of STAT5 decoy oligonucleotides (ODNs) on trans-activation of its downstream target bcl-x gene in K562 cells. METHODS: STAT5 decoy ODNs, mismatched ODNs (M-ODNs), and FAM-decoy ODNs were designed and synthesized. FAM decoy ODNs were used as control, and transfected into K562 cells by cationic liposomes, analyzed by flow cytometry (FCM) and fluorescent inversive microscopy. The bcl-x promoter fragment acquired by polymerase chain reaction (PCR) was inserted into pGL3-basic to construct luciferase report plasmid pGL3b-bclxp, which was co-transfected with decoy ODNs or M-ODNs into K562 cells. The activity of luciferase was detected. After transfection of decoy ODNs, and M-ODNs, expression of bcl-xL mRNA in K562 cells was detected by reverse transcription-PCR (RT-PCR),cell apoptosis was detected by FCM. RESULTS: FAM-decoy ODNs were incorporated into K562 cells in a dose-dependent manner. The incorporation efficiency reached 99.1% at the concentration of 4 micromol/L 24 h after transfection, and green fluorescence could be observed in cells under fluorescent inversive microscope. The luciferase activity was significantly lower in STAT5 decoy ODNs group than in control group [(181.48+/-204.46) RLU/microg protein vs. (675.26+/-62.91) RLU/mug protein, P < 0.05], but that of M-ODNs group [(632.07+/ 98.95) RLU/microg protein] has no significant difference with that of control group (P > 0.05). mRNA level of bcl-xL was decreased by STAT5 decoy ODNs, but not by M-ODNs. Moreover, Sub G1 peak was detected in STAT5 decoy ODNs group by FCM. CONCLUSION: Transfection of STAT5 decoy ODNs can down-regulate the trans activation of bcl-x in K562 cells. PMID- 15820063 TI - [In vitro anti-leukemia activity of cord blood original cytotoxic T lymphocytes]. AB - BACKGROUND & OBJECTIVE: Cord blood transplantation (CBT) possesses graft-versus leukemia (GVL) effect. It is possible to cure leukemia if specific cytotoxic T lymphocytes (CTLs) can be induced from lymphocytes in cord blood (CB), and be used to eradicate minimal residual disease (MRD) in leukemia patients. This study was to induce CTLs from cord blood, and explore its in vitro anti-leukemia activity. METHODS: Dendritic cells (DCs) were generated from 8 samples of cord blood mononuclear cells by culturing with multiple cytokines. Immature DCs were pulsed with apoptotic leukemia cells to present leukemic antigens to cord blood original lymphocytes to obtain CTLs. The characteristics of maturation of DCs were evaluated by morphology and flow cytometry. Anti-leukemia effect of CTLs was measured by lactate dehydrogenase release assay. RESULTS: Typical DCs were induced from all of the 8 samples. Expressions of immunologic markers CD1a(+), HLA-DR(+), CD86(+), CD83(+) on DCs were significantly higher after culturing than before culturing [(29.6+/-13.8)% vs. (1.8+/-1.9)%, (81.1+/-17.8)% vs. (19.4+/ 10.6)%, (42.7+/-21.9)% vs. (7.5+/-5.3)%, (8.0+/-6.9)% vs. (1.4+/-1.1)%, respectively, P< 0.05]. When E:T ratio was 50:1, CTLs showed far higher cytotoxicity to uncultured acute myeloid leukemia (AML) cells than to K562 cells, and to mononuclear cells of bone marrow from the corresponding patients in complete remission phase (CR-MNCs) [(52.6+/-21.0)% vs. (18.2+/-20.2)%, and (3.3+/ 6.3)%, P < 0.05]. CONCLUSIONS: Mature DCs derived from cord blood, which loaded leukemia antigens, could induce leukemia-specific CTLs. The CTLs have vigorous cytotoxicity to original leukemia cells rather than CR-MNCs. PMID- 15820064 TI - [In vitro inducing differentiation of bone marrow mononuclear cells of chronic myeloid leukemia]. AB - BACKGROUND & OBJECTIVE: Interferon-alpha (IFN-alpha), an important immunoregulatory cytokine, has been widely used in treating virus hepatitis, lymphoma, and chronic myeloid leukemia (CML), and showed evident effect, but the mechanism is unclear. Dendritic cells (DCs), specialized antigen-presenting cells (APCs), play a pivotal role in activating initial T cells, and maintaining cell immune responses. Does the efficiency of IFN to CML relate to the DCs induced by IFNy What kind of effect do DCs have on IFN therapy for CMLy Up to now, few researches are available. This study aimed to observe whether the DCs were induced through culturing bone marrow mononuclear cells (BMMNCs) of CML in vitro, investigate the mechanism of IFN-alpha therapy for CML, and then provide a new strategy for clinical therapy. METHODS: BMMNCs were obtained from blood of CML patients by Ficoll-Paque density gradient centrifugation, and induced with IFN alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) (IFN-alpha/ GM-CSF group), or interleukin-4 (IL-4) and GM-CSF (IL-4/ GM-CSF group), or IFN alpha, GM-CSF, and IL-4 (IFN-alpha/GM-CSF/IL-4 group) for 7 days. Morphology of BMMNCs was observed under transmissional and optical microscope. The phenotypes [CD1a, CD83, CD86, human leukocyte antigen (HLA)-ABC, HLA-DR, CD54] were assayed by flow cytometry (FCM). The mixed lymphocyte reaction(MLR) of DCs was evaluated by MTT assay. RESULTS: After inducements, BMMNCs showed typical dendritic projections, and highly expressed CD1a, CD83, CD86, HLA-ABC, HLA-DR, and CD54. Positive rates of HLA-ABC and HLA-DR were higher in IFN-alpha/ GM-CSF group and IFN-alpha/GM-CSF/IL-4 group than in IL-4/ GM-CSF group (P<0.05). Positive rate of CD86 and MLR were the highest in IFN-alpha/GM-CSF/IL-4 group (P<0.05). Positive rates of DC antigens and MLR in IFN-resistant group were significantly lower than those in newly diagnosed group and IFN-sensitive group (P<0.05), but positive rate of CD86 and MLR have no significant difference among 3 groups in the presence of IFN-alpha/GM-CSF/IL-4 (P>0.05). CONCLUSIONS: The BMMNCs of CML cultured in the presence of IFN-alpha and other cytokines can be induced into DCs with morphologic and immunophenotypic characteristics, overexpresses major histocompatibility complex (MHC) molecules, co-stimulatory molecules, and adhesion molecules, and have enhancing MLR. The possible mechanism of IFN-alpha therapy for CML may be relate to DCs. PMID- 15820065 TI - [Antitumor effects of chemotherapeutic drugs on fresh human gastric cancer cells and their relationships to expressions of P-glycoprotein and glutathione S transferase-pi]. AB - BACKGROUND & OBJECTIVE: Drug-resistance is a major factor of influencing treatment efficacy of advanced gastric cancer. This study was to evaluate in vitro antitumor effects of different chemotherapeutic drugs on fresh human gastric cancer cells, and explore their relationships with expressions of P glycoprotein (P-gp), and glutathione S transferase -pi (GST-pi) in human gastric cancer tissue. METHODS: A total of 39 specimens of highly purified gastric cancer cells were separately exposed to 5-fluorouracil (5-FU), cisplatin (DDP), mitomycin C (MMC), adriamycin (ADM), and hydroxycamptothecin (HCPT). Inhibitory rate of cells was detected by MTT assay. Metabolic activity of cells was detected by trypan blue exclusive assay. Cell apoptosis was detected by in situ terminal deoxynucleotidyl transferase assay (TdT assay). Expressions of GST-pi and P-gp in gastric cancer tissues were detected by immunohistochemistry. RESULTS: After exposure to antitumor drugs, morphologic changes, decrease of metabolic activity, and apoptosis were appeared in gastric cancer cells. Inhibitory rates of cancer cells exposed to MMC, DDP, 5-FU were significantly higher than those of cells exposed to ADM, and HCPT [(38.6+/-7.7)%, (38.1+/-8.8)%, and (37.8+/-10.3)% vs. (31.9+/-10.4)%, and (29.7+/-10.2)%, P < 0.01]. Apoptosis rate of cancer cells exposed to CDDP, 5-FU, and MMC were significantly higher than those of cells exposed to HCPT, and ADM [(32.1+/-7.7)%, (31.1+/-8.8)%, and (29.8+/-6.3)% vs. (21.9+/-7.4)%, and (19.9+/- 7.4)%, P < 0.05]. Positive rate of GST-pi was 66.7%(26/39), that of P-gp was 59.0%(23/39). GST-pi positive cells showed resistance to DDP and MMC, P-gp positive cells showed resistance to ADM and HCPT. CONCLUSIONS: Overexpressions of P-gp and GST-pi might contribute to drug resistance of tumor. Detection of GST-pi and P-gp, together with MTT chemosensitivity test, might be useful for selecting more effective chemotherapeutic drugs. PMID- 15820066 TI - [Radioprotective effect of aloe polysaccharides on three non-tumor cell lines]. AB - BACKGROUND & OBJECTIVE: Our previous study showed that aloe polysaccharides (AP) could evidently decrease the mortality of irradiated mice mainly through increasing the amount of hemocytes and ameliorating immune function of mice. Whether AP can protect the cells in vitro from irradiation damage is unknown. This study was to explore radioprotective effect of AP on 3 non-tumor cell lines, and its effect on cell cycle. METHODS: MTT assay was used to detect cytotoxicities of AP to normal human liver cell line Chang Liver (C. Liver), normal human embryo kidney cell line 293, and normal human umbilicus vein endothelial cell line ECV304. The 3 cell lines were treated with AP before or after irradiation. After 7-10 days normal culture, survival rate of cells was calculated by clone formation assay. Cell cycle was analyzed by flow cytometry (FCM) at different time points after irradiation. RESULTS: 293 cells were treated with AP at different time points before and after x-ray irradiation. Survival rate of 293 cells treated with AP 30 min before x-ray irradiation was the highest (64.2%) among all groups. Evident dosage-effect relationship of AP appeared in concentration range of 12.5-50 microg/ml. After treatment of 50 microg/ml of AP, survival rates of 293, ECV304, and C. Liver cells increased from 41.5%, 46.5%, and 40.9% to 49.4%, 72.1%, and 89.1%, respectively. Irradiation caused a distinct G(2)/M block and decreased G(0)/G(1) phase population in 293 and C. Liver cells. In C. Liver cells, pretreatment of 50 mug/ml of AP increased G(0)/G(1) phase population from 31.8% to 43.8%, decreased G(2)/M phase population from 38.5% to 13.8% 6 h after irradiation; and decreased G(2)/M phase population from 22.9% to 8.7% 24 h after irradiation. In 293 cells, the same pretreatment increased G(0)/G(1) phase population from 30.1% to 45.9% 6 h after irradiation, and from 40.4% to 45.2% 24 h after irradiation accompanied by decrease of G(2)/M population from 59.6% to 54.1%. CONCLUSIONS: AP has radioprotective effect on non tumor cells. This effect might relate to alleviating of cell cycle turbulence. PMID- 15820067 TI - [Inhibitory effect of cantharidin on invasion and metastasis of highly metastatic ovarian carcinoma cell line HO-8910PM]. AB - BACKGROUND & OBJECTIVE: Cantharidin, a natural toxin, has specific antitumor actions. Many researches confirmed that nuclear factor-kappaB (NF-kappaB) closely relates with invasion and metastasis of tumor. This study was designed to investigate inhibitory effect of cantharidin on metastasis-related ability of human highly metastatic ovarian carcinoma cell line HO-8910PM, and its mechanism. METHODS: MTT assay was used to examine cytotoxicity of cantharidin on HO-8910PM cells. Effect of cantharidin on adhesion potential of HO-8910PM cells was tested by cell-Matrigel adhesion assay. Transwell chamber assay was performed to determine its effect on invasion and migration capacities of HO-8910PM cells. Protein levels of NF-kappaB P65 subunit and vascular endothelial growth factor (VEGF) were assessed by Western blot. RESULTS: After 6-h treatment of 20 micromol/L of cantharidin, inhibitory rate of HO-8910PM cells was (8.4+/-2.2)%, inhibitory rates of invasion, migration, and adhesion capacities of HO-8910PM cells were (38.8+/-1.7)%, (40.3+/-5.6)%, and (55.1+/-6.7)%, respectively; protein levels of NF-kappaB P65 subunit and VEGF were down-regulated. CONCLUSIONS: Cantharidin can inhibit migration, invasion, and adhesion of HO-8910PM cells. Its possible mechanism may be involved in down-regulations of NF-kappaB P65 subunit and VEGF. PMID- 15820068 TI - [Inducement effect of (DIPP-L-Trp)2-L-Lys-OCH3 on apoptosis of K562 cells through mitochondrial-dependent pathway]. AB - BACKGROUND & OBJECTIVE: Phosphorylation of protein and peptide plays an important role in life activity. Some phosphoryl peptides are found to have activities to inhibit proliferation of tumor cells, but the involved mechanisms are unclear. This study was designed to investigate cell apoptosis induced by (O,O-diisopropyl phosphoryl-L-tryptophan)(2)-L-lysine methyl ester [(DIPP-L-Trp)(2)-L-Lys-OCH(3)], and its mechanism in K562 cells. METHODS: K562 cells were double stained by AnnexinV-FITC and propidium iodide (PI) to detect (DIPP-L-Trp)(2)-L-Lys-OCH(3) induced apoptosis by flow cytometry (FCM). After treatment of different concentrations of (DIPP-L-Trp)(2)-L-Lys-OCH(3), K562 cells were stained by rhodamine123 and PI to detect changes in membrane potential (Delta Psi m), or stained by 2',7'-dichlorofluorescein diacetate (DCFH-DA) to detect reactive oxygen species (ROS) of mitochondrial by FCM. RESULTS: When treated with 50 microg/ml of (DIPP-L-Trp)(2)-L-Lys-OCH(3) for 24 h, apoptosis rate of K562 cells was 61.9%, Delta Psi m was decreased in 93.6% of K562 cells,and ROS production was decreased. Both Delta Psi m and ROS production in K562 cells mitochondria were decreased with the increasing concentration and extending treatment time of (DIPP-L-Trp)(2)-L-Lys-OCH(3). CONCLUSION: (DIPP-L-Trp) (2)-L-Lys-OCH(2) could induce apoptosis in K562 cells, which might relate with down-regulation of mitochondrial Delta Psi m and reduction of ROS production. PMID- 15820069 TI - [Changes in apoptosis-related genes expression profile in human breast carcinoma cell line Bcap-37 induced by flavonoids from seed residues of Hippophae Rhamnoides L]. AB - BACKGROUND & OBJECTIVE: Hippophae rhamnoides L. possesses functions of antioxidation and radioprotection. This study was designed to investigate changes in apoptosis-related genes expression profile in human breast carcinoma cell line Bcap-37 induced by flavonoids from seed residues of Hippophae rhamnoides L. (FHR) with cDNA microarray, and to explore possible mechanism of signal transduction on apoptosis. METHODS: Total RNA was extracted from Bcap-37 cells before and after treatment of FHR. Two cDNA probes, labeled by Cy3-dUTP or Cy5-dUTP fluorescent dyes, were synthesized via reverse transcription, and hybridized with a microarray contained 13 824 human 14K cDNA. Differential gene expression profiles of FHR group and control group were analyzed by Genespring software. RESULTS: After treatment of FHR, 305 genes were up-regulated, and 361 were down-regulated; 32 apoptosis-related genes were differentially expressed, and accounted for 0.23% of the total genes in cDNA microarray. Of the 32 apoptosis-related genes, 25 were up-regulated (average Ratio: 3.071), and 7 were down-regulated (average Ratio: 0.418). Bioinformatic analyses showed that the 32 genes, including CTNNB1, TSSC3, IGFBP4, IGFBP6, GADD34, TNFRSF10B, Caspase-9, and PCNA, related with apoptosis of Bcap-37 cells when treated with FHR. CONCLUSION: Apoptosis of Bcap-37 cells induced by FHR relates with various genes through co-regulating of intracellular and extracellular signal transduction pathways. PMID- 15820070 TI - [Clinical randomized comparative trial of ProMACE-CytaBOM regimen and CHOP regimen in treating non-Hodgkin's lymphoma]. AB - BACKGROUND & OBJECTIVE: CHOP regimen is the routine modality for moderately or highly malignant non-Hodgkin's lymphoma (NHL). Previous reports show that ProMACE CytaBOM regimen could elevate complete response rate and survival rate of patients with moderately or highly malignant NHL. This study was to compare efficacies and safeties of ProMACE-CytaBOM and CHOP regimens in treating this disease, to indicate the standard treatment for it. METHODS: A total of 146 patients with moderately or highly malignant NHL, verified by pathology or histology, were randomized into trial group (73 patients treated with ProMACE CytaBOM regimen) and control group (73 patients treated with CHOP regimen). Survival rate was analyzed by Kaplan-Meier method. Chi-square test was performed between groups. RESULTS: The complete response rate, partial response rate, and response rate was significantly higher in ProMACE-CytaBOM group than in CHOP group (39.7% vs. 31.5%, 38.4% vs. 28.8%, and 78.1% vs. 60.3%, respectively, P < 0.05). The 1-, 3-, and 5-year survival rates were significantly higher in ProMACE CytaBOM group than in CHOP group (89.3% vs. 82.1%, 76.2% vs. 51.4%, and 45.7% vs. 32.3%, respectively, P < 0.05). The major side effects, appeared with no differences (P > 0.05) in incidences between 2 groups, were leukopenia, thrombocytopenia, and nausea. Each group had 1 case of treatment-related death. CONCLUSIONS: The results show higher efficacy of ProMACE-CytaBOM regimen over CHOP regimen. ProMACE-CytaBOM regimen may prolong survival time of patients with moderately or highly malignant NHL. PMID- 15820071 TI - [Treatment effect of DICE regimen on patients with relapsed or refractory intermediate and high grade non-Hodgkin's lymphoma]. AB - BACKGROUND & OBJECTIVE: So far, there is still no standard salvage regimen for relapsed or refractory non-Hodgkin's lymphoma (NHL). The response rates (RR) of NHL patients received common salvage regimens, such as DICE, ESHAP, MINE, and EPOCH, are only 30%-70%. This study was to evaluate the efficacy and safety of DICE regimen, as a salvage regimen, in treating patients with relapsed or refractory intermediate and high grade NHL. METHODS: Thirty-five patients with relapsed or refractory intermediate and high grade NHL, who had been pretreated with chemotherapy dominated by CHOP or CHOP-like regimen with a median of 6 cycles (ranged 2-12 cycles), were salvaged by DICE regimen from Jun. 1999 to Jan. 2004. Of the 35 patients, 14 were T-cell original, and 21 were B-cell original. RESULTS: The 35 patients received DICE regimen with a median of 4 cycles (ranged 2-7 cycles). All patients were assessable in the efficacy and adverse events. The total RR was 74.3% with complete response (CR) rate of 31.4%, median response time (MST) of 4 months (ranged 1-30 months), median time to failure (TTF) of 7 months (ranged 2-34 months),median survival time (MST) of 14 months (ranged 3-51 months), and 2-year survival rate of 33.3%. The RRs of T-cell and B-cell NHL were 85.7% and 66.7%. The CR rate was higher in T-cells NHL than in B-cell NHL (50.0% vs. 19.0%, P=0.073). Elevated serum lactate dehydrogenase (LDH) and bulky disease were high risk factors of the efficacy of DICE regimen (P < 0.05). The response to DICE reginmen was an independent prognostic factor of patients with relapsed or refractory NHL (P = 0.001). The major toxicity was myelosuppression. Incidences of neutropenia and thrombocytopenia of grade III-IV were 71.4% and 8.6%. CONCLUSIONS: DICE regimen is a safe and effective salvage regimen for the patients with relapsed or refractory intermediate and high grade advanced NHL. Elevated serum LDH and bulky disease are the adverse prognostic factors. The response to DICE regimen may directly influence survival time of patients with relapsed or refractory NHL. PMID- 15820072 TI - [Survival outcomes of T-cell non-Hodgkin's lymphoma: a report of 111 cases]. AB - BACKGROUND & OBJECTIVE: T-cell non-Hodgkin's lymphoma (NHL) is a group of heterogeneous malignancies with poor prognosis, and without ideal therapeutic regimen. This study was to summarize clinical and pathologic features of T-cell NHL. METHODS: Records of 111 patients with T-cell NHL, treated from Jan. 1994 to Dec. 2001 in Cancer Center of Sun Yat-sen University, were retrospectively analyzed. All the patients were classified according to WHO classification criteria. RESULTS: Median age of the whole group was 37 years (ranged 7-77 years). Of the 111 patients, 82 were men, 29 were women;45 (40.5%) were treated with chemoradiotherapy, 62 (55.8%) were treated with chemotherapy alone, and 4 (3.6%) were treated with radiotherapy alone. The 3-year survival rate of the whole group was 45% with a median follow-up of 28 months. The 3-year survival rates of chemoradiotherapy, chemotherapy, and radiotherapy groups were 56%, 38%, and 25%, respectively. Among all histological type subgroups, the prognosis of NK/T-cell lymphoma was the worst with the 3-year survival rate of only 25%u the 3 year survival rate was 40% in unspecified peripheral T-cell lymphoma group,and 85% in angioimmunoblast T-cell lymphoma group. International prognostic index was a significant factor for predicting overall survival. The 3-year survival rates of low risk,low-intermediate risk,intermediate-high risk, and high risk groups were 60%, 30%, 10%, and 0%, respectively. CONCLUSIONS: Present treatment modalities for T-cell NHL patients, especially the high risk patients, can't achieve satisfactory outcomes. New treatment modality for these patients needs to be explored. PMID- 15820073 TI - [Clinical analysis of 59 cases of primary gastric non-Hodgkin's lymphoma]. AB - BACKGROUND & OBJECTIVE: Gastrointestinal tract is the most common extranodal involvement site of non-Hodgkin's lymphoma (NHL). However, no standard treatment regimen has ever been established for primary gastric NHL (PGNHL). This paper was to summarize the clinical characteristics and treatment results of PGNHL patients. METHODS: Records of 59 PGNHL patients, treated from Jan. 1980 to Jan. 2002 in Cancer Center of Sun Yat-sen University, were reviewed to summarize their clinical characteristics, and influence of treatment modality on their survival. RESULTS: Of the 59 PGNHL patients, 46 (78.0%) were in stage I/II. According to Working Formulation, most of them were in intermediate grade (46, 78.0%). The most common immune phenotype was B-cell lineage (46/49, 93.9%). These patients were treated with chemotherapy plus surgery (37,62.7%), chemotherapy alone (17,28.8%), and surgery alone (5,8.5%), respectively. The 2-, 5-, and 10-year survival rates of the 59 patients were 76.4%,63.7%, and 42.5%, respectively. For those patients in intermediate grade (including immunoblastic cell lymphoma), there was no significant difference in the 5-year survival rate between the patients received chemotherapy plus surgery and the patients received chemotherapy alone (52.5% vs. 57.1%). CONCLUSIONS: Chemotherapy-dominated modality is recommended for patients with PGNHL of intermediate or high grade. The effect of surgery on PGNHL needs to be confirmed by prospective randomized trial. PMID- 15820074 TI - [Efficacy of intraoperative hypotonic peritoneal chemo-hyperthermia combined with early postoperative intraperitoneal chemotherapy on gastric cancer]. AB - BACKGROUND & OBJECTIVE: Abdominal recurrence from exfoliated cancer cells contributes a lot to treatment failure of advanced gastric cancer. Intraperitoneal chemotherapy, which has been proved effective in eliminating exfoliated cancer cells in abdominal cavity, is a hot topic on treatment of gastric cancer. This study was to explore application of combined therapy of intraoperative hypotonic peritoneal chemo-hyperthermia and early postoperative intraperitoneal chemotherapy to gastric cancer. METHODS: A total of 156 gastric cancer patients were randomized into 3 groups, and underwent the combined therapy (treatment group 1), intraoperative chemotherapy (treatment group 2), and peritoneal lavage with distilled water (control group), respectively. RESULTS: The 2-year survival rate of treatment group 1 was significantly higher than that of control group (88.4% vs. 65.2%, P < 0.05). The 3-year survival rate of treatment group 1 was significantly higher than those of treatment group 2, and control group (71.1% vs. 50.0%, and 45.6%, P < 0.05). Occurrence of liver metastasis was significantly lower in treatment groups 1 and 2 than in control group (7.7%, and 10.2% vs. 27.3%, P < 0.05). CONCLUSIONS: Combined therapy of intraoperative hypotonic chemo-hyperthermia and early postoperative intraperitoneal chemotherapy is effective for gastric cancer. Intraperitoneal chemotherapy can be used to prevent postoperative liver metastasis of gastric cancer. PMID- 15820075 TI - [Relationship of serum level of dihydropyrimidine dehydrogenase and serum concentration of 5-fluorouracil to treatment response and adverse events in colorectal cancer patients]. AB - BACKGROUND & OBJECTIVE: Toxicities and response are different when patients with advanced colorectal cancer were treated with standard FOLFOX6 regimen. Serum level of dihydropyrimidine dehydrogenase (DPD) relates to efficacy and toxicities of chemotherapy containing 5-fluorouracil (5-FU). This study was to explore relationship of DPD to serum concentration of 5-FU in colorectal cancer patients treated with FOLFOX6 regimen, and their correlation to treatment response and adverse events. METHODS: Serum level of DPD in 72 patients with colorectal cancer was detected by high-performance liquid chromatography (HPLC) before chemotherapy. Serum concentration of 5-FU at steady state was detected by HPLC after patients received FOLFOX6 regimen. Treatment response and adverse events in the patients were assessed. RESULTS: Serum levels of DPD were normally distributed in 72 patients (ranged 1.55-5.94), while serum concentrations of 5-FU at steady state were not (ranged 141.1-1 741.9 microg/L). Serum level of DPD was negatively correlated with serum concentration of 5-FU (r=-0.460, P < 0.01). Occurrence of adverse events was lower when 5-FU concentration was less than 600 microg/L than when 5-FU concentration was more than 600 microg/L (P < 0.05). The mean serum concentration of 5-FU was significantly higher in patients with complete response and partial response than in patients with steady disease, and progressive disease (513.9 microg/L vs. 409.8 microg/L, and 259.3 microg/L, P < 0.05). Serum level of DPD was lower in patients suffered oral mucositis and diarrhea of grade II-IV than in patients suffered oral mucositis and diarrhea of grade 0-I (P=0.016, P=0.047). Serum level of DPD had no relation with treatment response of the patients (r=0.312, P=0.078). CONCLUSIONS: DPD level and serum 5 FU concentration vary a lot among patients with colorectal cancer. DPD level negatively correlates with serum 5-FU concentration. Serum concentration of 5-FU correlates with treatment effect and toxicities. PMID- 15820076 TI - [Treatment effect of percutaneous vertebroplasty combined with interventional chemotherapy on vertebral metastases]. AB - BACKGROUND & OBJECTIVE: Vertebral metastasis is a common manifestation of patients with advanced cancer without effective treatment. This study was to explore treatment value and efficacy of percutaneous vertebroplasty (PVP) combined with interventional chemotherapy on vertebral metastases. METHODS: Seventy-five patients with vertebral metastases (42 men, 33 women; aged 31-76 years) were divided into 2 groups: 39 were treated by PVP combined with chemotherapy (VPCC group), and 36 were treated by PVP alone (VP group). All procedures were guided by CT scan. The results and complications were evaluated by pain questionnaire and routine follow-up. RESULTS: Response rate was significantly higher in VPCC group than in VP group (93.0% vs. 74.4%, P < 0.05); complete response rates of VPCC group and VP group were 25.6% and 10.3%. Common complication of VPCC was transient aggravation of pain. CONCLUSIONS: PVP may release the pain, and consolidate the vertebrae of patients with vertebral metastases. Its short-term effect may be enhanced by adding drugs into bone cement. PMID- 15820077 TI - [Drug concentration in axillary lymph nodes of patients with breast cancer after lymphatic chemotherapy]. AB - BACKGROUND & OBJECTIVE: Lymph node status is one of the decisive prognostic factors of breast cancer. Chemotherapy targeting regional lymphatic tissues has emerged as a promising therapy for malignancies with high tendency to disseminate lymphatically. This study was to detect drug concentration in axillary lymph nodes of patients with breast cancer after lymphatic chemotherapy (LC), and to investigate effect of LC on accumulation of antitumor drugs in regional lymph nodes through comparing with the effect of intravenous chemotherapy (VC). METHODS: Sixty patients with breast cancer, confirmed by preoperative puncture biopsy, were randomized into 2 groups, 30 (LC group) were subcutaneously injected with 4 ml of carboplatin-activated carbon suspension (containing 20 mg of carboplatin) around the primary tumor, the other 30 (VC group) were intravenously injected with an equal dose of aqueous carboplatin. Every 6 patients from each group received modified radical mastectomy 1, 12, 24, 36, or 48 h after injection. Axillary lymph nodes were removed for pathologic examination. The concentration of carboplatin in nodes was detected by Zeeman atomic absorption spectrometry. RESULTS: A total of 275 axillary lymph nodes were resected, with 154 in LC group and 121 in VC group. Of the 275 lymph nodes, 136(49.5%) were from 23 patients (38.3%) had pathologically detected metastases. The concentrations of carboplatin were significantly higher in LC group than in VC group 1, 12, 24, 36, and 48 h after injection [(11.82+/-3.50) microg/g vs. (0.06+/-0.02) microg/g, (23.58+/-7.34) microg/g vs. (0.11+/-0.05) microg/g, (18.22+/-4.93) microg/g vs. (0.10+/-0.02) microg/g, (16.70+/-5.15) microg/g vs. (0.05+/-0.02) microg/g, and (14.62+/-4.29) microg/g vs. 0, respectively, P < 0.001]. Lymph node metastasis had no correlation with drug concentration (P > 0.05). CONCLUSION: Compared with VC, LC can effectively and continuously improve drug concentration in axillary lymph nodes of patients with breast cancer. PMID- 15820078 TI - [Local failure-related factors of esophageal carcinoma after concurrent chemoradiotherapy]. AB - BACKGROUND & OBJECTIVE: Concurrent chemoradiotherapy is the standard treatment for unresectable esophageal carcinoma. Local failure and recurrence are main reasons of treatment failure. This study was to discuss the factors related with local failure and recurrence of unresectable esophageal carcinoma after concurrent chemoradiotherapy. METHODS: Records of 132 patients with esophageal carcinoma, treated with concurrent chemotherapy in our hospital from Jan. 1996 to Dec. 2003, were reviewed. The patients received the first cycle of chemotherapy at the beginning of radiotherapy, and the second cycle of chemotherapy when radiation dose escalated to 40 Gy. Local failure-related factors were analyzed by binary logistic regression. RESULTS: To the terminal of follow-up, of the 132 patients, 54 had local failure or in-fields recurrence, 20 had in-fields recurrence and lymph nodes or distant metastasis, 5 had out-fields recurrence. Logistic regression analysis showed that short-term effect and irradiation dose were correlated with local failure and recurrence. Local failure and recurrence rate was lower in patients with complete remission (CR) than in patients with partial remission (PR) after chemoradiotherapy (44.9% vs. 79.6%, P < 0.001); and the mean time to recurrence was longer in CR patients than in PR patients (12.9 months vs. 6.1 months, P=0.002). When irradiated with 50-60 Gy,60.1-69.9 Gy, >/=70 Gy of x-ray, the local failure rates of the patients were 69%, 61%, and 52% (P=0.027), respectively, the mean time to recurrence were 5.3, 9.1, and 10.3 months, respectively. CONCLUSION: Radiation dose and short-term effect may influence local control of unresectable esophageal carcinoma. PMID- 15820079 TI - [Effect of expression of coxsackie and adenovirus receptor on antitumor activity of genetically modified adenovirus]. AB - BACKGROUND & OBJECTIVE: The effect of adenovirus, a kind of oncolytic virus and a kind of carrier in gene therapy, depends on the expression of Coxsackie and adenovirus receptor (CAR) on cell surface to a large degree. This study was to evaluate relationship of CAR expression to infectivity and efficacy of adenovirus. METHODS: Pathologic specimens from 29 patients in a clinical trial of genetically modified adenovirus (H101) were collected. Expression of CAR in cancer tissues was detected by immuohistochemistry. Expression of CAR on cancer cell membrane was detected by flow cytometry (FCM). Inhibitory effect of H101 on cells was assessed by MTT assay. RESULTS: Positive rate of CAR was significantly higher in patients achieved complete remission (CR) or partial remission (PR) than in patients had stable disease (SD) or progressive disease (PD) [70.0% (7/10) vs. 31.6% (6/19), P=0.048]. CAR expression on different cells was different, and the amount of CAR was positively related to virus infectivity, presented as the inhibitory rate of H101-infected cells (r=0.986). CONCLUSIONS: The expression of CAR closely relates to antitumor activity and efficacy of adenovirus. Also, the efficacy of adenovirus is higher on cancers with increased CAR expression than on those with decreased CAR expression. PMID- 15820080 TI - [Relationship of serum level of creatine kinase of MB type to cardiac function of patients with advanced tumors and its prognostic value]. AB - BACKGROUND & OBJECTIVE: Patients with advanced tumors usually suffer from cardiac dysfunction. Monitoring cardiac function is important for these patients. Serum creatine kinase of MB type (CK-MB) is a biochemical marker of cardiac damage. This study was to estimate relationship of serum level of CK-MB to cardiac function of patients with advanced tumors, and its prognostic value. METHODS: Serum CK-MB in 68 patients with advanced tumors was detected by immunoinhibition assay. Observation group was composed of 34 patients with elevated serum CK-MB. Control group was composed of 34 patients with normal serum CK-MB level. RESULTS: Incidence of cardiac insufficiency of grade III-IV was significantly higher in observation group than in control group (21 vs. 6, P < 0.05). Mortality of observation group was significantly higher than that of control group (25 vs. 8, P < 0.01). Elevation of serum CK-MB was frequently found in patients with primary liver cancer or liver metastasis (P < 0.05). CONCLUSIONS: Patients with elevated serum CK-MB often suffer from cardiac insufficiency, severe illness status, and have high mortality. Serum level of CK-MB may be a predictor of poor prognosis of patients with advanced tumors. PMID- 15820081 TI - [Molecular mechanism of specific induction of apoptosis in tumor cells by apoptin]. AB - Apoptin, a small protein derived from chicken anemia virus, can specifically induce apoptosis in transformed cells or tumor cells, but not in normal cells. The tumor specificity of Apoptin relates to its subcellular localization. In transformed cells or tumor cells, Apoptin migrates to the nuclei, whereas in non transformed cells, it remains mainly within the cytoplasm. Phosphorylation is responsible for the nuclear localization of Apoptin. In tumor cells, Apoptin is phosphorylated, then translocates into the nuclei, and induces cell apoptosis. Apoptin-induced apoptosis does not depend on functional p53, and can't be inhibited by overexpression of Bcl-2 and Bcl-xL, but caspase-3 activation is necessary for Apoptin-induced rapid apoptosis. Apoptin has a strong tendency to aggregate, and exists as aggregates in living cells, but in vivo formation and dissociation of the aggregates are not required for apoptosis-inducing activity of Apoptin. It is possible that Apoptin's ability to bind DNA closely relates to its ability to induce apoptosis. PMID- 15820082 TI - DNA vaccine and asthma therapy. PMID- 15820083 TI - DNA vaccine encoding Der p 2 allergen generates immunologic protection in recombinant Der p 2 allergen-induced allergic airway inflammation mice model. AB - BACKGROUND: DNA immunization is a promising novel type of immunotherapy against allergy. An estimated 79.2% patients with asthma, wheezing and/or rhinitis suffer from Dermatophagoides pteronyssinus group 2 (Der p 2) allegen. The aim of the present study was to determine whether DNA vaccine encoding Der p 2 could generate immunologic protection in recombinant Der p 2 (rDer p 2) allergen induced allergic airway inflammation mice model and to understand the role of DNA vaccination in specific-allergen immunotherapy for asthma. METHODS: After DNA vaccination, BALB/c mice were sensitized by intraperitoneal injection (i.p) and challenged by intranasal instillation of rDer p 2. The lung tissues were assessed using hematoxylin and eosin. Mucus-producing goblet cells were identifed using periodic acid-Schiff (PAS)/alcian blue. The total cell number and composition of bronchoalveolar lavage samples were determined. The levels of the cytokines IL-4 and IFN-gamma, as well as IgE and IgG2a in the serum were determined by enzyme linked immunosorbent assay. Allergen-specific IL-4 and IFN-gamma production by spleen cells were also measured by enzyme-linked immunosorbent assay. Expression of signal transducer and activator of transcription 6 (STAT6) in splenocytes were determined by Western blot. RESULTS: DNA vaccine encoding Der p 2 allergen inhibited extensive infiltration of inflammatory cells and production of mucin induced by allergen. The influx of eosinophils into the lung interstitium was significantly reduced after administration of DNA vaccine. Significant reductions of IL-4 and increase in levels of IFN-gamma in bronchoalveolar lavage fluid were observed. The allergen-specific IgE was markedly decreased in mice receiving DNA vaccination. Allergen could induce higher IFN-gamma, weaker IL-4 in cultured spleen cells from mice receiving DNA vaccine. DNA vaccination inhibited STAT6 expression of spleen cells induced by allergen. CONCLUSION: These results indicated that DNA vaccine encoding Der p 2 allergen generates immunologic protection in recombinant Der p 2 allergen-induced allergic airway inflammation mice model with regulating the immune response towards a Th1-type reaction. PMID- 15820084 TI - Association of gene polymorphisms of tumour necrosis factor-alpha and interleukin 13 with chronic obstructive pulmonary disease in Han nationality in Beijing. AB - BACKGROUND: Genetic factors are believed to play a role in the individual susceptibility to chronic obstructive pulmonary disease (COPD). A single nucleotide polymorphism (SNP) of tumour necrosis factor-alpha (TNF-alpha) has been reported but inconsistent results may arise from different populations and phenotypes of COPD. There are only a few published studies of interleukin-13 (IL 13) SNPs on COPD. The SNPs of TNF-alpha and IL-13 have not been studied in the Chinese population. This research was conducted to study the frequencies of IL-13 gene promoter 1055 (IL-13-1055) and TNF-alpha gene-308 polymorphisms in the patients with COPD and to investigate the effect of those genetic polymorphisms on COPD in the Chinese population. METHODS: A cohort of COPD patients and age matched controls were recruited from an inpatient hospital service in Beijing. Venous blood was obtained and genomic DNA was extracted from peripheral blood monocytes using standard method. Genomic DNA was used as a template for amplification by polymerase chain reaction (PCR) to determine the polymorphism at -1055 in the IL-13 gene promoter region. PCR restriction fragment length polymorphism (RFLP) was used to determine polymorphisms in the TNF-alpha gene-308 position. The products were investigated by sequence analysis also. RESULTS: One hundred and eleven COPD patients and 97 controls were studied. Seventy-five cases were current smokers in COPD patients and 36 were current smokers in controls. The frequencies of TT genotype in the IL-13 gene promoter region were 11.7% (13/111) in the COPD group and 13.4% (13/97) in the controls (P = 0.713). However, the OR value of TT genotype was significantly increased to 6.4 (95% CI 1.62 - 25.39) in the smokers with COPD. TT genotype was also positively related to family history of COPD, OR = 7.7 (95% CI 1.37 - 43.80). The frequencies of A allele in the TNF-alpha gene were 5.9% in COPD and 3.1% in controls (P = 0.131). The OR value of A allele was 5.0 (95% CI 1.011 to 25.059) in smokers with COPD. CONCLUSIONS: There is no significant difference in the frequencies of the TT genotype of IL-13-1055 or the A allele of the TNF-alpha between Han Chinese patients with COPD versus control. Thus, it does not appear that these SNPs are independent factors in COPD for Han nationality in Beijing. However, these SNPs may increase the risk of COPD among smokers. PMID- 15820085 TI - Correlation between survivin mRNA expression and homoharringtonine induced apoptosis of malignant hematopoietic cells. AB - BACKGROUND: The inhibitor of apoptosis (IAP) gene family is involved in the suppression of apoptotic cell death as well as an increasing number of seemingly unrelated cellular functions. It is not known, however, whether IAP expression in malignant hematopoietic cells is affected by chemotherapeutic agents such as homoharringtonine (HHT). In this study, we investigated mRNA expression levels of IAPs, especially survivin, in various hematopoietic cell lines in relation with apoptosis induced by HHT. METHODS: Semiquantitative reverse transcriptase polymerase chain reaction was used to determine survivin mRNA levels. Cell apoptosis was examined by flow cytometry. Cell viability and proliferation assay was evaluated by MTT. The experiments were performed on the malignant hematopoietic cell lines MUTZ-1, K562, Jurkat, RMPI and HL60, with or without survivin antisense-oligodeoxynucleotides (AS-ODN) and HHT. RESULTS: The expression levels of survivin mRNA were variable in the cell lines and negatively correlated to HHT induced cell apoptosis. Survivin AS-ODN significantly decreased mRNA level of survivin, but not those of bax and bcl-2. Survivin also inhibited MUTZ-1 cell growth and induced apoptosis in a dose dependent manner. AS-ODN and HHT showed synergistic effect on MUTZ-1 cell growth. CONCLUSION: The apoptotic effect of HHT on the hematopoietic cell lines is associated with decreased level of survivin expression. Survivin could be a new marker for drug sensitivity and a new target for cancer treatment. PMID- 15820086 TI - Plasma level and genetic variation of apolipoprotein E in patients with lipoprotein glomerulopathy. AB - BACKGROUND: Lipoprotein glomerulopathy (LPG) is a renal disease characterized by thrombus-like lipoproteins in the glomerular capillaries and its abnormal lipoprotein profiles with marked elevation of apolipoprotein E (apoE). In this study, 15 Chinese patients with LPG were involed in exploring the association of the genetic variation and its plasma level in the pathogenesis of LPG. METHODS: A retrospective analysis of the clinical and pathological features was made in 15 patients with LPG. Plasma concentrations of apoE were measured with radial immunodiffusion assay. Genetic variations of apoE gene were detected using polymerase chain reaction and restriction fragment length polymorphism. Glomerular deposition of apoA, apoB and apoE in these patients were detected by immunofluorescence staining using monoclonal antibodies. RESULTS: Biochemical profiles of lipids and lipoproteins revealed markedly elevated levels of triglyceride, apoB and apoE, but approximately normal levels of total cholesterol, apoA1 and lipoprotein(a) [Lp(a)], which resembled familial hypertriglyceridemia. Genetic analysis demonstrated that the genotype distribution of apoE were 7 cases with epsilon3/epsilon4, 4 cases with epsilon3/epsilon3 and 2 cases with epsilon2/epsilon3. The other 2 cases (a mother and her son) showed a same distinct band. The band pattern of later 2 cases was quite similar to the apoE variant of Tokyo type. The calculated allele frequency of epsilon 4 was relatively high in cases with LPG in comparison with that in the normal controls. We further divided the 13 patients into three groups according to their genotypes of apoE. Patients with the genotype of apoE epsilon2/epsilon3 showed a lower level of plasma apoE as compared to those with apoE epsilon3/epsilon4 (P < 0.05). The serum level of high-density lipoprotein (HDL) was the lowest in patients with the genotype of apoE epsilon3/epsilon4. No difference was found among the patients with different apoE genotype in the other clinical and pathological characteristics. CONCLUSIONS: The genotype of apoE epsilon3/epsilon4 is the predominant one in Chinese patients with LPG. Patients with this genotype tend to have a higher plasma level of apoE and more severe lipid dysmetabolism. No correlation was found between the genotype of apoE and the clinical features in patients with LPG. PMID- 15820087 TI - Inhibition of vascular endothelial growth factor gene expression by T7-siRNAs in cultured human retinal pigment epithelial cells. AB - BACKGROUND: Retinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV. METHODS: Human primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence. RESULTS: Three pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7 sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol). CONCLUSION: T7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV. PMID- 15820088 TI - Potassium channels in airway smooth muscle and airway hyperreactivity in asthma. PMID- 15820089 TI - Studies of coltivirus in China. AB - OBJECTIVE: The purpose of this article is to review the developments of studies of Coltivirus in China. DATA SOURCES: The data used in this review was obtained mainly from the studies of Coltivirus reported from 1990 to 2003 in China. STUDY SELECTION: Relevant articles on studies of Coltivirus in domestic and foreign literature were selected. DATA EXTRACTION: Data were maily extracted from the articles which are listed in the reference section of this review. RESULTS: Many Coltiviruses have been isolated not only from blood samples of patients with unknown fever or from cerebrospinal fluid of patients with encephalitis in Xishuangbanna area in Yunnan province, but also from mosquitoes collected in many areas in China. In some patients diagnosed as Japanese encephalitis or unknown fever, an increase of Coltivirus IgG antibody of fourfold, or more, has been detected using ELISA. Similarly, Coltivirus IgM antibody was positive in some patients with Japanese encephalitis or viral encephalitis. From most Chinese patients, except the northeastern, the isolates of Coltiviruses belong to subgroup B2, according to RT-PCR amplification of the ninth and twelfth segments of the isolates and sequence analysis of their amplicons. Some biological properties of Chinese Coltiviruses isolates are different from that of North American Coltiviruses. CONCLUSIONS: The isolates of Coltiviruses from Chinese patients are one of the common agents causing viral encephalitis and unknown fever in summer-autumn season. It might be an important public health problem due to its high isolation rate and wide distribution in China. Mosquito is the main transmission vector of the virus. PMID- 15820090 TI - Effects of pinacidil on proliferation of cultured rabbit airway smooth muscle cells induced by endothelin-1. PMID- 15820091 TI - Effect of vascular endothelial growth factor and its receptor KDR on human airway smooth muscle cells proliferation. PMID- 15820092 TI - Comparison of antileukemic immunity mediated by dendritic cells derived from multidrug resistant leukemia K562/A02 cells and sensitive K562 cells. PMID- 15820093 TI - Renoprotective effect of combining angiotensin II receptor blockers and statins in diabetic rats. PMID- 15820094 TI - Protective effect of propofol at clinically relevant concentrations on primary cultured newborn rat hippocampus. PMID- 15820095 TI - Protective effect of cardiomyopeptidin on cultured rat hippocampal neurons injured by anoxia reoxygenation. PMID- 15820096 TI - A one-year clinical trial using didanosine, stavudine and nevirapine for highly active antiretroviral therapy. PMID- 15820097 TI - Coronary angiographies of patients with recurrent acute coronary syndrome following coronary artery bypass grafting. PMID- 15820098 TI - Percutaneous closure of ventricular septal defect associated with anomalous inferior vein cava drainage via transjugular approach. PMID- 15820099 TI - Anti-estrogen prevents xenoestrogen-induced testicular pathology of eelpout (Zoarces viviparus). AB - Estrogenic alkylphenols have been shown to affect the reproductive system of male fish causing induction of vitellogenin synthesis and altered testis structure. However, it is still unknown whether the histopathological effects on the testes is mediated by the estrogen receptor or if it represent general toxicopathological effects. In the present study, the effects of different concentrations of the estrogenic chemical 4-tert-octylphenol on vitellogenin (Vtg) synthesis and testicular structure were investigated in the eelpout Zoarces viviparus during spermatogenesis. Adult male eelpout were exposed to 4-tOP (nominal concentrations: 10, 50 or 100 microg l(-1)) or 17beta-estradiol (E2; 0.5 microg l(-1)) in a continuous flow-through system for 3 weeks. A group of fish were exposed to 4-tOP (50 microg l(-1)) concomitantly with the anti-estrogen ZM 189,154 (20 microg g(-1) week(-1), i.p.). The Vtg concentration in plasma was measured by enzyme-linked immunosorbent assay. The testicular structure was examined by light microscopy and gamma-glutamyl transpeptidase (gamma-GTP) activity was measured in the testes. The testicular localization of gamma-GTP was analysed by enzyme histochemistry. A marked increase in the plasma Vtg concentration was observed after exposure to the actual concentration of 35 microg l(-1) 4-tOP (nominal concentration, 50 microg l(-1)), 63 microg l(-1) 4 tOP (nominal concentration, 100 microg l(-1)) or E2. Co-treatment with ZM 189,154 totally abolished the 4-tOP-dependent induction of Vtg synthesis. Exposure to 4 tOP or E2 caused a marked reduction in the testis mass and severely affected the testicular development and structure including the Sertoli cells (based on histology and gamma-GTP activity), resulting in impairment of spermatogenesis and degeneration of lobular structures. Other cellular abnormalities such as accumulations of yellowish-brown pigmented cells and increased interstitial fibrosis in the testes was also observed in the exposed fish. In the groups exposed to the nominal concentrations of 50 or 100 microg l(-1) all fish had severely affected testes, while both normal, moderately and severely affected testes were found in the group exposed to the nominal concentration of 10 microg l(-1). Co-treatment with ZM 189,154 abolished part of these 4-tOP-induced effects on the testicular growth and histological structure. The study demonstrates that an anti-estrogen can abolish effects on the testis caused by estrogenic chemicals, providing evidence that some of the effects are mediated by the estrogen receptor. PMID- 15820100 TI - Parasitism in fish--an endocrine modulator of ecological relevance? AB - Wild bream (Abramis brama) were collected from the river Elbe, and the influences of parasitic infection by the tapeworm Ligula intestinalis on endocrine and related functions (vitellogenin [VTG]; plasma sex steroids: 17beta-estradiol [E2], 11-ketotestosterone [11-KT] and testosterone [T]; relative gonad [GSI] and liver [HSI] growth; maturation stages of germ cells [MS]; prominence of spawning tubercles [STI]) were investigated. Distinct regional differences in infection rates of bream with L. intestinalis were observed along the Elbe with the highest prevalences at the Czech border (up to 45%) and Magdeburg (up to 65%), areas that are heavily contaminated with complex mixtures of organic chemicals and metals. Parasitized fish of both sexes had significantly lower GSIs and poorly developed gonads (low MS). In males, a significant reduction in the prominence of spawning tubercles occurred. Infected females had significantly lower plasma VTG concentrations. A selective suppression of the sex steroids 11-KT and E2 was observed in male and female bream, respectively. Testosterone was not affected in the same manner in fish of both sexes. At sites with an elevated prevalence of L. intestinalis, the extent of the infection of an individual was significantly correlated with the suppression of the measured biomarkers. However, when applying a linear model to compare regional differences in infection prevalence with biological parameters, not all of the observed differences could be explained by parasitization by L. intestinalis. This indicates that other factors such as pollution may have contributed to the effects on reproductive and endocrine processes that occurred along the river. Given that sites with high prevalences of L. intestinalis were also characterized by elevated pollution, it is possible that there exists a combinatory effect of both pollution and parasitization that can have a serious impact on the reproductive capacity of a population, such as was observed at the sampling site at Magdeburg. PMID- 15820101 TI - Molecular mechanism of the aryl hydrocarbon receptor activation by the fungicide iprodione in rainbow trout (Oncorhynchus mykiss) hepatocytes. AB - The dicarboximide fungicide iprodione (Ip) causes oxidative damage as a result of the production of free oxygen radicals, and induces cytochrome P4501A3 (CYP1A3) in cultured rainbow trout hepatocytes. The aim of this study was to characterise some of the molecular mechanisms by means of which Ip activates the aryl hydrocarbon receptor (AhR) and subsequently induces the CYP1A3 gene in rainbow trout (Oncorhynchus mykiss). The study was performed using primary hepatocytes and transfected HepG2 cells with a reporter construct, in which luciferase gene expression is under the transcriptional control of a multimerised xenobiotic response elements (4XREs), or a 2.3 Kb DNA fragment (corresponding to the trout CYP1A3 gene promoter). Ip exposure increased rainbow trout hepatocyte CYP1A3 mRNA over time and increased the expression of reporter gene in HepG2, thus suggesting that Ip induces the CYP1A3 gene by activating the AhR. Genistein, a tyrosine kinase inhibitor, efficiently inhibited the Ip-mediated induction of the CYP1A3 gene as demonstrated by mRNA level decrease and the impaired activation of the luciferase reporter gene constructs. Staurosporine, an inhibitor of protein kinase C, also suppressed the induction by Ip. When the AhR antagonist alpha naphthoflavone was added to the cultures, Ip-mediated CYP1A3 induction was suppressed. These findings are consistent with a mechanism of Ip-mediated CYP1A3 gene induction that involves the activation of the AhR complex via phosphorylation-dephosphorylation reactions. PMID- 15820102 TI - Tentative biomarkers for 2,4,6-trinitrotoluene (TNT) in fish (Oncorhynchus mykiss). AB - 2,4,6-Trinitrotoluene (TNT) is the major explosive in ammunition dumped into lakes and the sea after World War II. To identify useful biomarkers of TNT exposure for forthcoming fish monitoring studies at ammunition dumping sites, rainbow trout (Oncorhynchus mykiss) were intraperitoneal (i.p.) injected with TNT in peanut oil at doses of 0, 100, 200 or 400 mg TNT/kg body weight and sampled 72 h later. The study covered blood parameters, and hepatic antioxidant and detoxifying enzymes. Fish treated with TNT had an increased glutathione S transferase (GST) activity and glutathione reductase (GR) activity, and a decreased percentage of oxidised glutathione (%GSSG) compared to the control group. In addition to increased methemoglobin, the increased glutathione and glutathione dependent enzyme activities indicate that TNT oxidises macromolecules and activates antioxidant defence systems which may be useful as general biomarkers of TNT-exposure. The fish bile was analysed for TNT and its metabolites by gas chromatography-mass spectrometry (GC-MS), and the toxicity of the bile was determined with the cladoceran Ceriodaphnia dubia. A dose-dependent increase in TNT, 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6 dinitrotoluene (4-ADNT) was found in the hydrolysed bile of the TNT-treated fish. These results indicate that the fish are able to detoxify and excrete TNT and suggest that the detection of TNT, 2-ADNT and 4-ADNT in bile may be suitable as a direct marker of exposure to TNT. PMID- 15820103 TI - Do copper tolerant fathead minnows produce copper tolerant adult offspring? AB - The objective of this study was to determine if the relative Cu tolerance of fathead minnow parents determines the relative Cu tolerance of their adult offspring. It was hypothesized that the adult offspring of Cu-tolerant minnows would inherit Cu tolerance from their parents. The relative Cu tolerance of 96 adult fish was determined based upon their reduction in swim performance following a sublethal exposure to 150 microg Cu/l. Control, Cu-tolerant and Cu susceptible lines of fish were produced and fish within each line were allowed to breed. The offspring were raised to adults, then exposed to one of two sublethal Cu concentrations (150 or 225 microg Cu/l) for 8 days. There were no significant differences in relative Cu tolerance, as measured by reduction in swim performance, among the three lines of fish at either dose. However, significant differences in whole body Na+ occurred among the fish lines after exposure to 150 microg Cu/l, but not after exposure to 225 microg Cu/l. Significant differences in whole body Cu occurred between Cu-tolerant and Cu-susceptible fish lines after exposure to either Cu dose. The offspring did not inherit the relative Cu tolerance of their parents, however, the selection lines had diverged from each other, particularly with respect to their whole body Cu concentrations after exposure. PMID- 15820104 TI - Short-term effects of endocrine-disrupting chemicals on the expression of estrogen-responsive genes in male medaka (Oryzias latipes). AB - To evaluate the estrogenic activities of selected estrogenic compounds such as estradiol-17beta (E2), nonylphenol (NP), 4-(1-adamantyl)phenol (AdP), bisphenol A (BPA), BPA metabolite 4-methyl-2,4-bis(4-hydroxyphenyl)pent-1-ene (MBP) and 4,4' dihydroxy-alpha-methylstilbene (DHMS) in the shortest possible time, we investigated the expression of estrogen-responsive genes such as vitellogenin I, vitellogenin II and alpha-type estrogen receptor genes in the liver of male medaka (Oryzias latipes) using reverse transcription-polymerase chain reaction (RT-PCR) techniques. These estrogen-responsive genes responded rapidly to selected estrogenic compounds after 8 h exposure, and the expression of hepatic vitellogenin II and estrogen receptor alpha mRNA was found to be more responsive than that of vitellogenin I mRNA. As a result, the relative estrogenic potencies of tested chemicals descended in the order of E2 (100)>MBP (0.38)>AdP (0.25)>DHMS (0.05)>NP (0.02)>BPA (0.001). Moreover, this preliminary study indicates that AdP and DHMS should be considered as candidate estrogenic compounds with the potential to induce hepatic estrogen-responsive genes in male medaka. These results suggest that vitellogenin I, vitellogenin II and estrogen receptor alpha gene expression patterns alter in male medaka treated with selected estrogenic compounds, and that these genes may be useful molecular biomarkers for screening estrogenic endocrine-disrupting chemicals in the shortest possible time. PMID- 15820105 TI - Bioaccumulation of waterborne selenium in the Asiatic clam Corbicula fluminea: influence of feeding-induced ventilatory activity and selenium species. AB - A set of experiments was performed to investigate the bioavailability and the effect of Se on the ventilatory activity of the bivalve Corbicula fluminea, under different conditions of both algal cell densities and dissolved Se chemical forms and concentrations. A first set of experiments was conducted without selenium to investigate the changes in the ventilatory flow rate as a function of the concentration of the unicellular alga Chlamydomonas reinhardtii (10(5)-10(6) cells mL(-1)). For algal concentrations below 2-3x10(5) cells mL(-1), ventilatory activity was highly stimulated whereas it was independent of algal densities for higher values (up to 10(6) cells mL(-1)). To investigate the influence of this first ventilatory drive on selenium contamination process, bivalves were exposed to waterborne selenium at two different algal concentrations, selected to provide contrasting reference ventilatory activities. Three different selenium forms were studied [selenite Se(+IV), selenate Se(+VI) and selenomethionine SeMet] and were added into the water at concentrations of 50 and/or 500 microg L(-1). Each selenium form induced a specific behavioural response, an increase, a decrease or no change of ventilation being observed for Se(+IV), SeMet and Se(+VI), respectively. Selenium accumulation by the organisms was investigated at the organ level for the different exposure conditions. Selenomethionine was the most bioaccumulated form, followed by selenate and selenite, respectively. Despite the bivalves displaying different ventilatory behaviours at low or high algal density, there was no evidence showing reduction or enhancement of Se uptake in the chemical domain investigated. PMID- 15820106 TI - Toxic cyanobacterial cells containing microcystins induce oxidative stress in exposed tilapia fish (Oreochromis sp.) under laboratory conditions. AB - The effects of microcystins from cyanobacterial cells on various oxidative stress biomarkers in liver, kidney and gill tissues in freshwater tilapia fish (Oreochromis sp.) were investigated under laboratory conditions. Microcystins are a family of cyclic peptide toxins produced by species of freshwater cyanobacteria (blue-green algae). Fish were exposed to the cyanobacterial cells in two ways: mixed with a commercial fish food or crushed into a commercial fish food so that the toxins were released. Two different exposure times were studied: 14 and 21 days. The oxidative status of fish was evaluated by analyzing the level of lipid peroxidation (LPO), as well as the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR). The findings of the present investigation show that microcystins induce oxidative stress in a time-dependent manner and that the type of administration of the cyanobacterial cells influences the extent of these effects. Thus, the crushed cyanobacterial cells (released toxins) induced the antioxidant defences studied and increased the LPO level to a greater extent than the non-crushed cells. The liver was the most affected organ followed by kidney and gills. These results together with reports that fish can accumulate microcystins mean that cyanobacterial blooms are an important health, environmental and economic problem. PMID- 15820107 TI - Experimental results on bioaccumulation of metals and organic contaminants from marine sediments. AB - A test-system for the assessment of bioavailability and bioaccumulation of metals and organic contaminants in marine benthic organisms is described and results from studies where this system has been applied are assessed. Sediments tested were polluted harbour sediment (from Norway), and clean sediments spikes with metal containing weight materials for drilling muds. Contaminants that may bioaccumulate under relevant conditions are indicated. The test-system uses two species of ecological relevance (Nereis diversicolor and Hinia reticulata). Interspecies differences in bioaccumulation were found for several compounds, which show the importance of using species with different modes of living in such tests. Compared to other PAHs, pyrene was found to bioaccumulate to a high degree (BioAccumulation Ratio, BAR=213.5>sediment concentration ratio, SCR=97.4; bioaccumulation factor, organism dw. conc. to sediment dw. conc., BAF=1.02), which shows that extrapolating bioaccumulation results between different substances is difficult. When assessing bioavailability of specific compounds, it is most adequate to perform direct measurements on exposed organisms, such as the experiments described here. The high bioaccumulation of compounds such as pyrene and nickel may in some cases be attributed to manipulation of the sediments and (for pyrene) lack of subsequent aging, thereby overestimating bioavailability. PMID- 15820108 TI - The non-linear effect (determined by the penalised partial-likelihood approach) of milk-protein concentration on time to first insemination in Belgian dairy cows. AB - The time to first insemination in dairy cows depends partly on the energy balance of the cow. Because milk-protein concentration is related to the energy balance, we investigated whether milk-protein concentration predicted the hazard of being inseminated. The main objective of the paper is to demonstrate that the relationship between milk-protein concentration and the hazard of being inseminated was not linear and that this non-linear relationship was modelled adequately using cubic-splines. The semiparametric Cox model was used to introduce protein concentration into the model as a time-varying covariate and additionally herd was added to the model as a frailty term to adjust for the clustering of the cows within a herd. We extended the penalised partial likelihood technique to fit the frailty model with cubic-splines for the effect of the protein concentration. The model was fitted for a large database consisting of 5114 multiparous cows from 181 different farms. Low milk-protein concentration (<2.7%) was associated with a negative energy balance and this probably led to the decreased hazard. On the other hand, high milk-protein concentration (>4.0%) was linked with low milk production and it was probably a farmer's decision not to inseminate such cows, leading to the observed decreased hazard. PMID- 15820109 TI - Factors associated with time to neonatal diarrhoea in French beef calves. AB - A prospective study was carried out on 92 randomly selected beef herds in the Midi-Pyrenees region in France. The objective was to determine factors associated with time to neonatal gastroenteritis. By taking into account the "intra-herd" correlation in failure time (in the semiparametric Cox model), we identified 12 management risk factors associated with hazard of diarrhoea. Some previously have been identified, but "new" risk factors were feeding of corn silage and the incidence of diarrhoea in the last season. We used the two main approaches which are often reviewed: marginal and frailty Cox models. Our results show that these two models give different parameter estimates, so the choice of the model remains crucial. PMID- 15820110 TI - Active circulation of bluetongue vaccine virus serotype-2 among unvaccinated cattle in central Italy. AB - Several seroconversions occurring in 2002 among sentinel cattle during the bluetongue-vaccination campaign in Lazio and Tuscany (central Italy) led to the suspicion of vaccine-virus circulation. Therefore in 2003, 17 seroconverting sentinel herds were investigated for the characteristics of the virus involved. From these farms, 91 unvaccinated animals and 57 Culicoides pools were tested for the presence of the bluetongue vaccine virus (serotype-2) or other strains. The presence of vaccine virus serotype-2 was confirmed by PCR followed by restriction analysis in the whole blood of 17 unvaccinated sentinel cattle and 12 pools of Culicoides imicola or C. obsoletus. Of the 17 herds, five were positive only for vaccine virus serotype-2, four were positive for other strains and two for both the vaccine and other strains; the remaining premises were virologicaly negative. The vaccine virus serotype-2 also was detected in areas not included in the vaccination campaign. PMID- 15820111 TI - Monte Carlo simulation of animal-product violations incurred by air passengers at an international airport in Taiwan. AB - Our aim was to evaluate the inspection of arriving passengers for the illegal transport of animal products through the international airport into Taiwan. Passenger-violation data were retrieved from the Bureau of Animal and Plant Health Inspection and Quarantine (BAPHIQ) in Taiwan. Monte Carlo simulations with 5000 iterations were applied to estimate the predicted violation numbers considering factors such as passenger's nationality/ethnicity, sex, and monthly statistics of violations. The results showed that Chinese-related passengers and passengers from South East Asian countries have a relatively higher potential risk for incurring violations during the period from Christmas to Chinese Lunar New Year. Most monthly estimated violation risks are significantly higher than observed violation risks in 2001 and 2002, except April, July, and August. Due to the low sensitivity of the current detection system (reflected by the higher estimated violation risks and a two-fold detection increase after the implementation of an inspection service using detector dogs), we recommend additional preventive measures be taken. PMID- 15820112 TI - Herd-level risk factors for infectious diseases in Swedish dairy calves aged 0-90 days. AB - The effect of environmental factors and management routines on the risk of diarrhoea, respiratory disease and other infectious diseases was investigated in 3081 heifer calves 0-90 days old in 122 Swedish dairy herds. The farmers kept records on cases of diseases in their heifer calves and in addition, project veterinarians clinically examined all calves every 2-3 months. At each visit, the veterinarians also measured the ammonia concentration and relative air humidity in the housing facilities for the calves. The cleanliness of the animals and their environment was recorded as a measure of the hygienic status of the farm. The presence or absence of draught (i.e. wind velocity>0.5 m/s) was recorded twice during the study period. The effect of these factors, as well as the placing of the calf pens, the nature of the pen walls, air volume per animal, management factors (such as the status of the caretaker and feeding routines) and presence or absence of a bovine viral diarrhoea virus (BVDV) infection in the herd, was evaluated by means of a two-level variance component logistic model. The placing of calf pens along an outer wall was significantly associated with the risk of diarrhoea (odds ratio (OR): 1.92, P<0.01). The risk for respiratory disease was significantly associated with an ammonia concentration below 6 ppm (OR: 0.42, P<0.05) while the odds ratio for moderately to severely increased respiratory sounds was significantly associated with a BVDV infection in the herd (OR: 2.39, P<0.05) and draught (OR: 3.7, P<0.02). Absence of draught was significantly associated with the risk for infectious diseases other than diarrhoea and respiratory disease (OR: 0.42, P<0.01). PMID- 15820113 TI - Estimation of diagnostic-test sensitivity and specificity through Bayesian modeling. AB - We review recent Bayesian approaches to estimation (based on cross-sectional sampling designs) of the sensitivity and specificity of one or more diagnostic tests. Our primary goal is to provide veterinary researchers with a concise presentation of the computational aspects involved in using the Bayesian framework for test evaluation. We consider estimation of diagnostic-test sensitivity and specificity in the following settings: (i) one test in one population, (ii) two conditionally independent tests in two or more populations, (iii) two correlated tests in two or more populations, and (iv) three tests in two or more populations, where two tests are correlated but jointly independent of the third test. For each scenario, we describe a Bayesian model that incorporates parameters of interest. The WinBUGS code used to fit each model, which is available at http://www.epi.ucdavis.edu/diagnos-tictests/, can be altered readily to conform to different data. PMID- 15820114 TI - Factors associated with variation in bulk-tank-milk Salmonella Dublin ELISA ODC% in dairy herds. AB - Our objective was to determine factors that contribute to variation in bulk-tank milk Salmonella Dublin enzyme-linked immunosorbent assays (ELISA) corrected optic density measurements (ODC%) in dairy herds. We constructed hierarchical mixed models with repeated bulk-tank-milk ELISA ODC% in 31 Danish dairy herds. Four models included different combinations of explanatory factors, and we compared how well these models described the variation in the data. Herd was included as a random effect nested within Salmonella status and barn type. Detection of Salmonella Dublin or Salmonella Typhimurium by bacteriological culture of individual faecal samples or of slurry samples was associated with higher bulk tank-milk ELISA ODC%, as was apparent Salmonella prevalence, the mean ELISA ODC% or mean-yield-corrected ELISA ODC% in milk samples collected from all individual cows. However, combinations of risk factors that included number or prevalence of cows with a very high ELISA ODC% provided better models, indicating that the effect of the cow-level explanatory variables on the bulk-tank-milk ELISA ODC% was related to the activity of the infection in the herd. Barn type (loose housing or tie stalls) was not associated with the variation in bulk-tank-milk ELISA ODC% in these models, which might be useful in planning of surveillance programs and intervention strategies. PMID- 15820115 TI - Economic impacts of reduced pork production associated with the diagnosis of Actinobacillus pleuropneumoniae on grower/finisher swine operations in the United States. AB - An examination of the economic impacts of the diagnosis of Actinobacillus pleuropneumoniae on grower/finisher swine operations indicated that reduced pork production, associated with the diagnosis of A. pleuropneumoniae on the operation, diminished consumer surplus by $53+/-52 million, and resulted in a total loss of $32+/-30 million to the US economy in 1995. Most of the economic surplus lost by consumers was transferred to producers, whose economic surplus increased by $21+/-25 million (which was not significantly different from zero). Uncertainty analysis showed that an estimate of the decline in production associated with the diagnosis of A. pleuropneumoniae accounted for most of the uncertainty of the change in consumer surplus and of the total loss to the economy. The estimate of the price elasticity of demand for pork also contributed towards a lot of the uncertainty in the estimated change in producer surplus. PMID- 15820116 TI - Persistence of elevated rabies prevention costs following post-epizootic declines in rates of rabies among raccoons (Procyon lotor). AB - Determining the benefits to cost relationships among different approaches to rabies control and prevention has been hindered by the inherent temporal variability in the dynamics of disease among wildlife reservoir hosts and a tangible and objective measure of the cost of rabies prevention. A major and unavoidable component of rabies prevention programs involves diagnostic testing of animals and the subsequent initiation of appropriate public health responses. The unit cost per negative and positive diagnostic test outcome can be reasonably estimated. This metric when linked to methodologies subdividing the epizootic process into distinct temporal stages provided the requisite detail to estimate benefits derived from rabies control strategies. Oral rabies vaccine (ORV), for prevention of the raccoon-associated variant of rabies, has been distributed in Ohio and adjoining states in an effort to develop an immune barrier to the westward spread of epizootic raccoon rabies. The costs of ORV delivery have been quantified. Herein, the cost structures required to assess the benefits accrued by prevention were developed. A regression model was developed effectively predicting (r2=0.70) the total number of rabies diagnostic tests performed by 53 counties in five northeastern (NE) states from 1992 to 2001. Five temporal stages sufficed to capture the range of variability in the raccoon rabies epizootic process. Unit costs, dollars per diagnostic test outcome, were calculated for negative and positive results from published reports. Ohio counties were matched to NE counties based on similar socioeconomic characters. A "pseudo-epizootic" of raccoon rabies was introduced into Ohio and the costs savings from ORV were derived as the excess costs imposed by epizootic spread throughout the state. At 46 km/year (range modeled, 30-60 km/year), the pseudo epizootic spread, and reached the enzootic stage, in all Ohio counties by year 13 (range modeled, 11-17 years). Cumulative excess costs for Ohio ranged between $11 and $21 million; counties of low socioeconomic status experienced the greatest relative excess costs. The costs for rabies prevention activities reached apices during the epizootic stage of raccoon rabies (2.7-10.8 times baseline) an unforeseen finding indicated elevated costs persisted (1.7-7.2 times baseline) into the enzootic stage. PMID- 15820117 TI - Predicting the probability of abortion in dairy cows: a hierarchical Bayesian logistic-survival model using sequential pregnancy data. AB - Although abortion contributes substantially to poor reproductive health of dairy herds, little is known about the predictability of abortion based on age, previous abortion or gravidity (number of previous pregnancies). A poor understanding of effects of maternal factors on abortion risk exists, in part, because of methodological difficulties related to non-independence of multiple pregnancies of the same cow in analysis of fetal survival data. We prospectively examined sequential pregnancies to investigate relationships between fetal survival and putative dam risk factors for 2991 abortions from 24,706 pregnancies of 13,145 cows in nine California dairy herds. Relative risks and predicted probabilities of abortion (PPA) were estimated using a previously described hierarchical Bayesian logistic-survival model generalized to incorporate longitudinal data of multiple pregnancies from a single cow. The PPA increased with increasing dam age at conception, with increasing number of previous abortions, and if the previous pregnancy was aborted >60 days in gestation. The PPA decreased with increasing gravidity and with increasing number of days open. For cows that aborted, the median time to fetal death decreased slightly as gravidity increased. The study considers several methodological issues faced in epidemiologic investigations of fetal health, including multi-modal hazard functions, extensive censoring and non-independence of multiple pregnancies. The model improves our ability to predict bovine abortion and to characterize fetal survival, which have important applications to herd health management. PMID- 15820118 TI - Survival analysis on aggregate data to assess time to sero-conversion after experimental infection with Bovine Leukemia virus. AB - Bovine Leukemia virus (BLV) is a ubiquitous retrovirus that affects mainly cattle. Knowledge of the precise moment of infection is fundamental for identification and evaluation of factors related to BLV transmission. Systematic reviews and meta-analyses provide good evidence on the effects of medical interventions. The objectives were to estimate time to sero-conversion after experimental infection using data from retrieved literature and to detect factors that may influence the length of that interval using survival analysis on pooled data. An analysis using aggregate data from 36 studies totalling 438 observations was performed. From this, four sets were created and analysed by interval censored accelerated failure time models (AFT) with different distributions (exponential, Weibull, log-logistic, lognormal and generalized gamma), and some variants of the Cox model (Andersen-Gill, smoothing splines) with and without a frailty effect. The AFT gamma model fit best and the estimated median time to sero-conversion in the null model was 57 days (95% confidence interval (CI): 49; 75) using all data and 47 days (95% CI: 39; 55) when only studies using experimental inoculation were considered. Some factors were consistently associated with time to sero-conversion. These included exposure by animal-to animal contact (resulting in a seven-fold increase in time to sero-conversion compared to direct inoculation), diagnostic method to detect sero-conversion (time to sero-conversion was 1.4 times shorter when AGID was used compared to ELISA), and transmission by insect bites (biological media) delayed sero conversion 2.3 times compared transmission via needles or other inanimate media. After fitting a frailty Cox model, results showed that sero-conversion in susceptible animals after infection using donors, in which presence of virus before the experiment started was confirmed, increased the hazard of sero conversion two times in comparison with donors in which virus presence was not confirmed before start of the experiment. Inoculation with blood decreased the hazard 2.5 times in comparison with lymphocyte suspensions. Heterogeneity due to different research groups was also present. Finally, a Cox model with smoothing splines contained three variables: research group, route of inoculation and a non linear spline for infective dose. In conclusion, it can be stated some factors that influence the time to sero-conversion were identified and quantified and that a moderate influence of research centre existed. These results may contribute to the estimation of the most probable times of infection in field conditions and in a better evaluation of control measures. PMID- 15820119 TI - A quantitative assessment of the BSE risk associated with fly ash and slag from the incineration of meat-and-bone meal in a gas-fired power plant in Denmark. AB - It has been recommended that meat-and-bone meal (MBM) be incinerated at 850 degrees C for at least 2s and the ashes and slag disposed of in controlled landfills, to dispose of animal-derived proteins. Most commonly, the MBM is incinerated in cement works or coal-fired power plants and the ashes and slag are incorporated into the cement or concrete. Our goal was to assess with a Monte Carlo simulation model the bovine spongiform encephalopathy (BSE) risk to cattle and humans posed by the ash and slag. The results will be used by decision makers to evaluate the need for disposal of the fly ash in controlled landfills and the feasibility of use of the ash by the phosphate and fertilizer industries. We assumed that all specified risk material (SRM) and MBM produced in Denmark would be incinerated in this gas-fired power plant. Based on observations in 2001, we assumed that, on average, six (range: 0-15) clinical BSE cases each year were rendered into MBM and incinerated. In addition, SRM or carcasses from 0 to 31 (median=10) BSE-infected-but-undetected animals/BSE case were also incinerated. The simulations were run on a 1-week basis. Our results suggest that if the slag is collected and re-incinerated the median BSE infectivity remaining in the fly ash per week would be 3.1E-11 cattle ID(50). A cattle ID50 is the amount of infectivity that will cause infection in 50% of cattle exposed to it. During the weeks when BSE was infected in the SRM-MBM, the median infectivity in the fly ash was estimated as 8.7E-10 cattle ID50 and 2.9E-12 human ID50. The 95th percentiles were 2.1E-08 cattle ID50 and 5.8E-10 human ID50, respectively. One ton of fly ash would contain 0.05). Phagocytic and phenoloxidase activities were significantly higher in abalone fed diets supplemented with 800 mg PN kg(-1) than those fed the PN-free diet or the one with 4-deoxypyridoxine (P < 0.05). Agglutination titre and respiratory burst activity were significantly higher in abalone fed diets supplemented with 40 mg PN kg(-1) than those fed the PN-free diet or the one with 4-deoxypyridoxine (P < 0.05). There were no significant differences in immunological characteristics between the abalone fed the diet containing 40 mg PN kg(-1) and those fed the diet containing 800 mg PN kg(-1) (P > 0.05). L. japonica resulted in significantly lower agglutination titre, respiratory burst and phagocytic activities than the artificial diets supplemented with 40 or 800 mg PN kg(-1) (P < 0.05). Total haemocyte count (THC), serum protein concentration, and the activities of lysozyme and acid phosphatase were not significantly affected by the dietary treatments (P > 0.05). These results demonstrate that dietary deficiency of pyridoxine suppresses the immune functions in H. discus hannai, and further investigations are needed to optimise the dietary level of this vitamin for maintaining the best immune responses in abalone. PMID- 15820126 TI - The rodlet cells of teleostean fish: their potential role in host defence in relation to the role of mast cells/eosinophilic granule cells. AB - The distribution and potential function of the rodlet cells of teleosts were studied by microscopic observations on tissue samples from the digestive tract and adjacent tissues, including the bulbus arteriosus. Fish representing 3-5 genera from each of the families Salmonidae, Cyprinidae, Gadidae and Labridae were included in the study. Great individual variations in the distribution of rodlet cells were found in all species of salmonids, gadids and labrids. The cells seemed to be absent in some individuals of a species and were associated with different epithelial tissues in others, but were not found in vascular endothelia. Their occurrence was common in all salmonids caught in their natural environment, whereas those in aquaculture, kept under controlled conditions with respect to water quality, showed extremely few rodlet cells. In species of the cyprinid family, the picture was different. Rodlet cells were consistently present under the endothelium of the bulbus arteriosus, and were very numerous at this location in individuals infected with blood flukes. In other epithelial tissues of cyprinids, rodlet cells were encountered in fairly high numbers, but in some tissues of individuals from all species they were occasionally absent. In all of the studied families rodlet cells seemed to be recruited when helminths affected epithelial tissues. Mast cells/eosinophilic granule cells were consistently very numerous in tissues of the intestine of cyprinids and labrids. In gadids, mast cells/eosinophilic granule cells seemed to be absent. Present evidence points to a role for the rodlet cells in defence functions, e.g. in combating helminths, and the suggestion earlier made for mast cells/eosinophilic granule cells, that evolution has created a "standing force" in particular tissues of teleosts consistently exposed to pathogens, whereas an efficient "mobilization force" has been an advantage in those living in more pathogen-free environments, may also be applied to rodlet cells, explaining the differences between teleostean families with respect to their distribution pattern. PMID- 15820127 TI - Dopamine depresses the immune ability and increases susceptibility to Lactococcus garvieae in the freshwater giant prawn, Macrobrachium rosenbergii. AB - The total haemocyte count (THC), phenoloxidase activity, respiratory burst (release of superoxide anion), superoxide dismutase activity, and phagocytic activity and clearance efficiency to the pathogen Lactococcus garvieae were measured when freshwater giant prawns Macrobrachium rosenbergii (16.2 +/- 2.1 g) were individually injected with saline, or dopamine at 0.5, 5.0, or 50.0 pmol prawn(-1). The results show that a transient period of immunosuppression occurred between 2 and 8 h after injection of dopamine for all immune parameters except circulating haemocytes and all immune parameters returned to control values within 8-16 h after receiving dopamine. Injection of dopamine also significantly increased the mortality of M. rosenbergii challenged with the pathogen L. garvieae. These results suggest that stress-inducing dopamine suppresses the immune system, which in turn promotes the susceptibility to L. garvieae in M. rosenbergii. PMID- 15820128 TI - Dietary yeast RNA supplementation reduces mortality by Aeromonas hydrophila in rohu (Labeo rohita L.) juveniles. AB - A feeding trial was conducted for 60 days to delineate the effect of dietary ribonucleic acid or chitin on haematological parameters, phagocyte respiratory burst and resistance to Aeromonas hydrophila of Labeo rohita juveniles. One hundred and twenty-six (avg. wt. 13.40 +/- 0.17 g) juveniles were randomly distributed in six treatment groups, each one in three replicates. Six isonitrogenous (crude protein: 34.34-35.37%) and isocaloric (414-425 kcal 100 g( 1)) purified diets were prepared with different concentrations of either ribonucleic acid or chitin except the control group, viz., control, T1 (0.1% ribonucleic acid), T2 (0.2% ribonucleic acid), T3 (0.4% ribonucleic acid), T4 (25 mg chitin kg (-1)) and T5 (50 mg chitin kg (-1)). Weight gain %, specific growth rate, feed efficiency ratio, protein efficiency ratio did not vary significantly (P > 0.05) among the experimental groups. Haemoglobin content and total erythrocyte count were observed within the normal range and were not influenced by the dietary immunostimulants. Highest total leukocyte count was found in the T(3) group. The immunomodulatory effects of dietary immunostimulants were studied by using nitroblue tetrazolium (NBT) assay and serum parameters, namely total protein, albumin, globulin and A/G ratio. The respiratory burst activity (NBT) of blood phagocytes was highest in the T3 group followed by the T2 group, which varied significantly (P < 0.05) from other groups. Significantly (P < 0.05) higher total protein, globulin and lower A/G ratio was observed in the T(3) group. The relative percent survival after challenging with Aeromonas hydrophila was highest in the T3 group, compared to the control group, followed by the T2 group. The results indicate that dietary ribonucleic acid at 0.4% enhances phagocyte respiratory burst and protection of Labeo rohita juveniles to challenge by A. hydrophila. PMID- 15820129 TI - Cloning and nucleotide sequence of a novel 28-kDa protein from the mantle muscle of the squid Todarodes pacificus with homology to tropomyosin. AB - In recent studies, we found autodegradation of collagen from the mantle muscle of the squid Todarodes pacificus and also that the 28- and 25-kDa proteins are closely related to this phenomenon [Connect. Tissue Res. 45 (2004) 109-121]. We obtained partial sequences of three internal portions of this protein, which suggested that 25-kDa protein is a partially degraded form of the 28-kDa protein. We determined the full cDNA sequence of this protein by the degenerate polymerase chain reaction (PCR) using the information of amino acid sequences. The deduced amino acid sequence corresponding to the 212-bp cDNA contained all of the amino acid identified from the 28-kDa protein. Rapid amplification of cDNA ends (RACE) and squid mantle muscle RNA allowed cloning of the full 522-bp sequence, corresponding to a protein of 174 amino acids. A database search indicated that this is a new protein that shares 27-34% identity with tropomyosins from various animals. Structural prediction suggested that it possesses heptad repeats that form coiled-coil structures. We expressed a recombinant protein encoded by the 212-bp cDNA in Escherichia coli and used it to generate a polyclonal antibody. Western blotting with this antibody showed that the 28-kDa protein is expressed in fin, tentacle, and mantle muscle, but not in liver. PMID- 15820130 TI - Cytoskeletal ultrastructure and lipid composition of I-Z-I fraction in muscle from pre- and post-spawned female hake (Merluccius hubbsi). AB - Myofibrils from pre- and post-spawned female hake were depleted of thin and thick filaments by KI-treatment and the cytoskeletal ultrastructure of the preparations was investigated by electron microscopy. Irrespective of the gonadal stage of the fish, transmission (TEM) and scanning (SEM) electron micrographs showed an extensive network of filaments connecting Z-structures, which appeared as two circular rings held together. Z-structures of KI-treated myofibrils from post spawned hake were more compact than those from pre-spawned fish. This ultrastructural difference was absent when KI-treated myofibrils from hake in both gonadal conditions were prepared in the presence of a proteinase inhibitor cocktail (1 mM EDTA+1 mM PMSF+1 mM iodoacetic acid). The total lipid (TL) content of the I-Z-I fraction from fish in pre-spawned condition was about 3.7 mg/g I-Z-I and non-polar lipids (NPL) represent 86.5% of TL. TL and NPL contents of the I-Z I fraction from post-spawned hake were about 50% and 78.2% lower than those obtained from this fraction in pre-spawning fish. No significant changes (p>0.05) were observed in phospholipids (PL) and acidic phospholipids (AP). No significant differences (p<0.01) were observed among the corresponding lipid fractions of I-Z I purified from pre- and post-spawning fish in the presence of a protease inhibitor cocktail. PMID- 15820131 TI - Activation of oxidative burst and degranulation of porcine neutrophils by a homologous spleen galectin-1 compared to N-formyl-L-methionyl-L-leucyl-L phenylalanine and phorbol 12-myristate 13-acetate. AB - Galectins are a family of animal lectins defined by their beta-galactoside binding activities and a consensus sequence in their carbohydrate-recognizing domain (CRD). Relevant roles of galectins are described in adaptive immune response, innate immunity and modulation of the acute inflammatory response. We have extended our previous studies on a porcine spleen galectin-1 in relation to its functional roles such as polymorphonuclear neutrophils (PMNs) stimulation compared to well known PMN activators e.g. N-formyl-L-methionyl-L leucyl-L phenylalanine (fMLP) and phorbol 12-myristate 13-acetate (PMA). Relative to activation of NADPH-oxidase fMLP and PMA are stronger than galectin-1 plus cytochalasin B (CB) when the lectin is employed at low concentrations (gal-1 1 microM, 3.6+0.8 nm O(2)(-)/min/10(7) PMN). Higher doses of galectin-1 (10 microM) plus CB produced a significant activation of NADPH-oxidase (27.9+14.8 nm O(2)( )/min/10(7) PMN) and stimulated PMN degranulation up to 50%. We propose that local galectin-1 concentrations under physiological conditions might reach suitable levels for pig PMN stimulation, and might be a natural inducer of O(2)( ) formation or degranulation. Porcine galectins might produce enhanced responses in vivo when they stimulate neutrophils in combination with some other stimuli. PMID- 15820132 TI - Cysteine and serine protease-mediated proteolysis in body homogenate of a zooplankter, Moina macrocopa, is inhibited by the toxic cyanobacterium, Microcystis aeruginosa PCC7806. AB - The paper describes the characterization of proteases in the whole body homogenate of Moina macrocopa, which can possibly be inhibited by the extracts of Microcystis aeruginosa PCC7806. With the use of oligopeptide substrates and specific inhibitors, we detected the activities of trypsin, chymotrypsin, elastase and cysteine protease. Cysteine protease, the predominant enzyme behind proteolysis of a natural substrate, casein, was partially purified by gel filtration. The substrate SDS-polyacrylamide gel electrophoresis of body homogenate revealed the presence of nine bands of proteases (17-72 kDa). The apparent molecular mass of an exclusive cysteine protease was 60 kDa, whereas of trypsin, it was 17-24 kDa. An extract of M. aeruginosa PCC7806 significantly inhibited the activities of trypsin, chymotrypsin and cysteine protease in M. macrocopa body homogenate at estimated IC(50) of 6- to 79-microg dry mass mL(-1). Upon fractionation by C-18 solid-phase extraction, 60% methanolic elute contained all the protease inhibitors, and these metabolites could be further separated by reverse-phase liquid chromatography. The metabolites inhibitory to M. macrocopa proteases also inhibited the corresponding class of proteases of mammalian/plant origin. The study suggests that protease inhibition may contribute to chemical interaction of cyanobacteria and crustacean zooplankton. PMID- 15820133 TI - Glucose uptake in vivo in skeletal muscles of insulin-injected chicks. AB - Glucose uptake across the plasma membrane in animal cells plays a crucial role in whole-body glucose homeostasis. Insulin-stimulated glucose transport activity in vivo in several tissues was estimated using the 2-deoxy-D-[1-(3)H]glucose ([(3)H]2DG) uptake determination method. A tracer dose of [(3)H]2DG was injected intravenously into 8-day-old chicks (Gallus gallus) administered simultaneously or previously with porcine insulin (40 microg/kg BW). After 10 or 20 min, several major tissues, including skeletal and cardiac muscle, were sampled and their 2 deoxy-D-[1-(3)H]glucose 6-phosphate content analyzed. Plasma glucose concentration and [(3)H]2DG radioactivity were lowered by insulin within 20 min of [(3)H]2DG administration, while the plasma [(3)H]2DG/glucose ratio was not significantly different between chicks injected with insulin and their control counterparts. A marked uptake of 2DG was observed in cardiac tissue and brain, followed by kidney and skeletal muscles. In skeletal muscles, insulin increased the 2DG uptake in soleus, extensor digitorum longus and pectoralis superficialis muscles. On the other hand, no significant increases in insulin-induced 2DG uptake were detected in cardiac muscle or adipose tissue compared to controls. The results show that glucose transport across the plasma membrane in vivo in most skeletal muscles tested, but not cardiac muscle, was increased by insulin administration to chicks. These findings suggest that an insulin-responsive glucose transport mechanism is present in chickens, even though they intrinsically lack GLUT4 homologous gene, the insulin-responsive glucose transporter in mammals. PMID- 15820134 TI - Molecular cloning of cytochrome P450 aromatases in the protogynous wrasse, Halichoeres tenuispinis. AB - P450 aromatase (P450arom, CYP19), a CYP19 gene product, is a member of the cytochrome P450 superfamily that catalyzes the formation of aromatic C(18) estrogen from C(19) androgen. To begin to understand the molecular mechanisms of P450 aromatase action in the protogynous wrasse, we isolated two cDNAs: one encoding CYP19a from ovary and the other encoding CYP19b from brain. The full length cDNA of wrasse CYP19a, isolated from ovary cDNA library, is 2020 bp long and encodes 519 amino acids. The amino acid sequence of CYP19a has 62-83% identity with ovary-type aromatases of other teleosts. The full-length cDNA of wrasse CYP19b obtained using 5' and 3' RACE consists of 2666 bp, and its open reading frame encodes 496 amino acids. The deduced amino acid sequence has 62-83% identity with brain-type aromatases of other teleosts. Northern blot analysis identified a single 2.2-kb transcript in the ovary (CYP19a), and a single 2.6-kb transcript in the brain (CYP19b), suggesting that there are single forms of CYP19a and CYP19b, respectively, in the wrasse. RT-PCR assay showed that two CYP19 genes were expressed ubiquitously in various tissues, although each CYP19 subtype was expressed at highest level in the ovary and brain of the wrasse. These results suggest that CYP19 genes act in diverse tissue types, in addition to their effects on the physiological and reproductive functions of estrogen. PMID- 15820135 TI - Androgen receptor in the oviduct of the turtle, Trachemys scripta. AB - Circulating androgens reach high concentrations in females of some reptiles and amphibians. We are testing the hypothesis that androgens can act directly in female reptilian reproductive tissues, via the androgen receptor. In this study, we sought to determine if androgen receptors are present in the oviduct of the turtle, Trachemys scripta, using radioligand-binding assays and immunological assays. An androgen-binding site was detected in turtle oviductal cytosol and oviductal nuclear extract by radioligand binding assay, using (3)H dihydrotestosterone (DHT) as the ligand. This site was saturable (B(max)=11 pmol/g tissue), had a high affinity (10(-10) M), and showed specificity typical of androgen receptors (DHT>testosterone, progesterone>>estradiol, cortisol). Western blotting using an anti-androgen receptor antibody revealed a band of immunoreactivity in oviductal cytosol at approximately 115 kDa, and a more prominent band at 50 kDa, possibly indicating a truncated form of the androgen receptor. Immunohistochemistry revealed crossreactivity of the androgen receptor antibody against oviductal glandular cells but not against oviductal luminal epithelial or muscularis cells. The presence of androgen receptor in the turtle oviduct suggests that androgens have a role in female reproduction and that their action can be mediated directly by androgen receptor. PMID- 15820136 TI - Phylogenetic analysis and developmental expression of brp-like genes in amphioxus and zebrafish. AB - The brp-like genes in amphioxus (Branchiostoma belcheri tsingtauense) and in zebrafish (Danio rerio) are reported. The putative brp-like proteins are orthologous to the ancestor of the human brain protein 239FB and 239AB. Previous studies showed that human brain protein 239 might play a role in central nervous system development and function. In this study, the transcripts of zebrafish brp like gene are mainly located in the developing central neural system in embryo and larva similar to its orthologous genes in human. In contrast, the developmental expression pattern of amphioxus brp-like gene suggests that the gene might be involved in the development of the notochord, pharynx, gills and gut in amphioxus embryos and larvae. Phylogenetic analysis shows that the amphioxus brp-like gene is closer to the orthologous genes in vertebrates than those in invertebrates. The results suggest that the function of the brain protein 239 family genes may change in evolution. PMID- 15820137 TI - Enzyme activities of intestinal triacylglycerol and phosphatidylcholine biosynthesis in Atlantic salmon (Salmo salar L.). AB - The substitution of fish oil with plant-derived oil in diets for carnivorous fish, such as Atlantic salmon, has previously revealed the potentially deleterious supranuclear accumulation of lipid droplets in intestinal cells (enterocytes) which may compromise gut integrity, and consequently, fish health. This suggests that unfamiliar dietary lipid sources may have a significant impact on intestinal lipid metabolism, however, the mode of lipid resynthesis is largely unknown in teleost fish intestine. The present study aimed at characterising three key lipogenic enzymes involved in the biosynthesis of triacylglycerol (TAG) and phosphatidylcholine (PC) in Atlantic salmon enterocytes: monoacylglycerol acyltransferase (MGAT), diacylglycerol acyltransferase (DGAT), and diacylglycerol cholinephosphotransferase (CPT). Furthermore, to investigate the dietary effect of plant oils on these enzymes, two experimental groups of fish were fed a diet with either capelin (fish oil) or vegetable oil (rapeseed oil:palm oil:linseed oil, 55:30:15 w/w) as the lipid source. The monoacylglycerol (MAG) pathway was highly active in the intestinal mucosa of Atlantic salmon as demonstrated by MGAT activity (7 nmol [1-(14)C]palmitoyl-CoA incorporated min(-1) mg protein(-1)) and DGAT activity (4 nmol [1-(14)C]palmitoyl-CoA incorporated min(-1) mg protein( 1)), with MGAT appearing to also provide adequate production of sn-1,2 diacylglycerol for potential utilisation in PC synthesis via CPT activity (0.4 nmol CDP-[(14)C]choline incorporated min(-1) mg protein(-1)). Both DGAT and CPT specific activity values were comparable to reported mammalian equivalents, although MGAT activity was lower. Nevertheless, MGAT appeared not to be the rate limiting step in salmon intestinal TAG synthesis. The homology between piscine and mammalian enzymes was established by similar stimulation and inhibition profiles by a variety of tested cofactors and isomeric substrates. The low dietary n-3/n-6 PUFA ratio presented in the vegetable oil diet did not significantly affect the activities of MGAT, DGAT, or CPT under optimised assay conditions, or in vivo intestinal mucosa lipid class composition, when compared to a standard fish oil diet. PMID- 15820138 TI - Mass spectral analysis of pig (Sus scrofa) apo HDL: Identification of pig apoA II, a dimeric apolipoprotein. AB - Comparative studies of mammalian high density lipoproteins have clearly indicated that the major apolipoprotein is apoA-I and in some mammals apoA-II is the second major apolipoprotein. However, in pigs, apoA-II has been considered to be either present in trace amounts or absent. Recently, cDNA sequences for pigs A-II have been entered into the database. Translation of these sequences revealed that pig A-II consisted of 77 amino acids and that a cysteine residue was at residue 6. The A-II of three other mammals, chimpanzees, horses and humans, also has a cysteine residue at this position. As a result of a disulfide bond formed between monomers, the A-II in each of these cases circulates as a homodimer. Using electrospray-ionization mass spectrometry (ESI-MS), we obtained molecular mass data demonstrating that dimeric apoA-II is also present in pig plasma. In addition to being the first to report on the presence of apoA-II in pig plasma, we also obtained values for the molecular masses of apoA-I, apoC-III, apoD and serum amyloid A protein. PMID- 15820139 TI - Stearoyl-CoA desaturase expression and fatty acid composition in milkfish (Chanos chanos) and grass carp (Ctenopharyngodon idella) during cold acclimation. AB - Desaturation of fatty acids is an important adaptation mechanism for fish to maintain membrane fluidity under thermal stress. To comprehend the temperature adaptation mechanism in fish, we investigated the difference in the changes of stearoyl-CoA desaturase expression and fatty acid composition between milkfish and grass carp under cold acclimation. We find that in both fish the proportions of unsaturated fatty acids at 15 degrees C are all higher than those at 25 degrees C. In milkfish Delta(9)-desaturation index (ratios of 16:1/16:0 and 18:1/18:0) increases significantly in the beginning of cold acclimation at 15 degrees C and decreases afterward, but in grass carp it increases slightly in the beginning of cold acclimation followed by a sustained dramatic increase. Similarly, activity of stearoyl-CoA desaturase in milkfish increases significantly in the beginning, peaks at day 4, and then decreases constantly, but in grass carp it increases gradually in the first week, rises dramatically afterward, and then maintains a very high level. The change of stearoyl-CoA desaturase activity is parallel to the change of Delta(9)-desaturation index in both milkfish and grass carp, but it is one day earlier than Delta(9) desaturation index in milkfish. The difference of adaptation capability between milkfish and grass carp under cold stress is further evidenced by RT-PCR and Northern blot analysis of stearoyl-CoA desaturase gene expression. PMID- 15820140 TI - A novel cysteine protease inhibitor with lectin activity from the epidermis of the Japanese eel Anguilla japonica. AB - A novel cysteine protease inhibitor (Eel-CPI-1) was isolated from the epidermis of the eel. Eel-CPI-1 was shown to bind strongly to both lactose- and carboxymethylated papain-affinity gels. Its molecular mass under reducing condition was determined to be 18 kDa by SDS-polyacrylamide gel electrophoresis but approximately 30.5 kDa under non-reducing-conditions. Eel-CPI-1 inhibited papain (K(i)=18 nM) and ficin (K(i)=120 nM) competitively. Combined with the data on amino acid and sequence analysis, Eel-CPI-1 is identical to the eel lectin, AJL-2. This is the first report describing a cysteine protease inhibitor with lectin activity. PMID- 15820141 TI - Characterisation of proteins in the milk of fur seals. AB - Milk protein composition was investigated throughout the lactation periods of the Australian fur seal (Arctocephalus pusillus doriferus) and Antarctic fur seal (Arctocephalus gazella). The mean protein content of the milk was found to be 10.9% and 10.6% respectively. The concentration of total protein did not change during lactation, although a decline in casein content of the milk in late lactation was apparent. Milk protein concentration during a foraging/suckling cycle of the Antarctic fur seal analysed at the time of arrival on shore, and 24 h and 72 h after arrival was 12.8%, 11.4% and 12.5% respectively. Re-feeding animals at 72 h resulted in a significant increase in milk protein content to 14.9%. Characterisation of milk protein by SDS-PAGE analysis revealed 5 casein and 10 major whey protein bands. Amino-terminal sequencing indicated that the majority of the whey fraction of the milk is beta-lactoglobulin (beta-LG). The limited amino acid sequence indicated 3 different beta-LGs were secreted in the milk. Subsequently, RT-PCR was used to extend the sequence of one of the beta-LGs and translation of the 464 bp fragment indicated that it shared 79% sequence identity with feline beta-LG II. PMID- 15820142 TI - Partial purification and characterization of pro-phospholipase A2 activating proteases from gill membranes of the red sea bream, Chrysophrys major. AB - We previously reported that gill group IB secretory phospholipase A(2) (sPLA(2)) exists as an inactive pro-sPLA(2) with the dipeptide Ala-Arg, at the N-terminus of mature sPLA(2) in mucous cells. Pro-sPLA(2) should be activated after being secreted to the surface of gill epithelia by trypsin-like protease. To clarify the above hypothesis, we investigated the existence of pro-sPLA(2) activating protease (PAP) in the gills of the red sea bream, using gill pro-sPLA(2) as a substrate. PAP was solubilized from the membrane fraction of the gills with 2% sodium cholate and partially purified by benzamidine-Sepharose chromatography and reversed-phase HPLC. Partially purified proteases, PAP1 and PAP2 showed a high molecular mass of about 200 kDa by gelatin zymography. PAP1 and PAP2 had optimal pH from 7 to 9 and were inhibited by trypsin inhibitors. These properties of PAP1 and PAP2 suggest that both enzymes belong to the membrane-associated trypsin-like serine protease family, such as enteropeptidase and corin. This is the first report verifying the existence of the activating protease of group IB pro-sPLA(2) isoforms in a non-digestive tissue. PMID- 15820143 TI - Transgene copy number-dependent rescue of murine beta-globin knockout mice carrying a 183 kb human beta-globin BAC genomic fragment. AB - We report the generation and characterisation of the first transgenic mice exclusively expressing normal human beta-globin ((hu)beta-globin) from a 183 kb genomic fragment. Four independent lines were generated, each containing 2-6 copies of the (hu)beta-globin locus at a single integration site. Steady state levels of (hu)beta-globin protein were dependent on transgene copy number, but independent of the site of integration. Hemizygosity for the transgene on a heterozygous knockout background ((hu)beta(+/0), (mu)beta(th-3/+)) complemented fully the hematological abnormalities associated with the heterozygous knockout mutation in all four lines. Importantly, the rescue of the embryonic lethal phenotype that is characteristic of homozygosity for the knockout mutation was also demonstrated in two transgenic lines that were homozygous for two copies of the (hu)beta-globin locus, and in one transgenic line, which was hemizygous for six copies of the (hu)beta-globin locus. Our results illustrate the importance of transgene copy number determination and of the hemizygosity/homozygosity status in phenotypic complementation studies of transgenic mice containing large heterologous transgenes. Transgenic mouse colonies with 100% (hu)beta-globin production from the intact (hu)beta-globin locus have been established and will be invaluable in comparative and gene therapy studies with mouse models containing specific beta-thalassemia mutations in the (hu)beta-globin locus. PMID- 15820144 TI - Quantitative change in mitochondrial DNA content in various mouse tissues during aging. AB - In order to systematically characterize age-related changes in the mtDNA content of various tissues during aging, we analyzed the mtDNA content of eight tissues from mice at five different ages from young to senescent by quantitative real time PCR analysis. Obvious variations of mtDNA content among the tissues were detected: There was a 20-fold range in 2-week-old mice and a 50-fold range in 15 month-old mice. The mtDNA contents of the heart, lung, kidney, spleen and skeletal muscle increased gradually with age, whereas those of bone marrow and brain showed no age-related pattern. The expression patterns of mitochondrial transcription factor A (mtTFA) and mitochondrial single-strand DNA binding protein (mtSSB), possible regulatory factors of the mtDNA copy number, were not necessarily linked with the age-related pattern of the mtDNA content, suggesting the existence of other factors that affect the mtDNA content. The Western blot analysis of mtDNA-encoded cytochrome c oxidase subunit III (MTCO3) demonstrated that the expression levels of this protein in the heart and skeletal muscle increase with age in parallel with the mtDNA content. These findings confirm that the mtDNA content of tissues changes during aging. PMID- 15820145 TI - Collaborative action of M-CSF and CTGF/CCN2 in articular chondrocytes: possible regenerative roles in articular cartilage metabolism. AB - It is known that expression of the macrophage colony-stimulating factor (M-CSF) gene is induced in articular chondrocytes upon inflammation. However, the functional role of M-CSF in cartilage has been unclear. In this study, we describe possible roles of M-CSF in the protection and maintenance of the articular cartilage based on the results of experiments using human chondrocytic cells and rat primary chondrocytes. Connective tissue growth factor (CTGF/CCN2) is known to be a potent molecule to regenerate damaged cartilage by promoting the growth and differentiation of articular chondrocytes. Here, we uncovered the fact that M-CSF induced the mRNA expression of the ctgf/ccn2 gene in those cells. Enhanced production of CTGF/CCN2 protein by M-CSF was also confirmed. Furthermore, M-CSF could autoactivate the m-csf gene, forming a positive feed back network to amplify and prolong the observed effects. Finally, promotion of proteoglycan synthesis was observed by the addition of M-CSF. These findings taken together indicate novel roles of M-CSF in articular cartilage metabolism in collaboration with CTGF/CCN2, particularly during an inflammatory response. Such roles of M-CSF were further supported by the distribution of M-CSF producing chondrocytes in experimentally induced rat osteoarthritis cartilage in vivo. PMID- 15820146 TI - Three-dimensional reconstruction of chick calvarial osteocytes and their cell processes using confocal microscopy. AB - Osteocytes are surrounded by hard bone matrix. Therefore, it has not previously been possible to demonstrate the real architecture of the osteocyte network in bone. We previously reported that it is possible to observe osteocytes in bone by labeling the cells with fluorescence and using confocal laser scanning (CLS) microscopy. In this study, we for the first time conducted an extensive analysis of the morphology and morphometry of the three-dimensional (3D) osteocyte structure using three-dimensionally reconstructed fluorescent images. Sixteen-day old embryonic chick calvariae were stained with fluorescently labeled phalloidin and observed using a confocal laser scanning microscope. Morphometry of osteocytes in the calvaria was analyzed using extensive three-dimensional reconstructing software IMARIS, process length measuring software NEURON TRACER and cell surface area-/cell volume-analyzing software SURPASS. From the IMARIS derived images, we found that the average of 10 osteocytes is 52.7 +/- 5.7 processes, and the point-to-point distance between centers of the osteocytes was 24.1 +/- 2.8 microm. In addition, we could calculate that each osteocyte spans an average of 4180 +/- 673 microm3 of bone volume. NEURON TRACER showed that the length of osteocyte processes was 0.26 +/- 0.02 microm per 1 microm3 bone compartment. In addition, SURPASS indicated that the surface area of osteocytes was 0.36 +/- 0.03 microm2 per 1 microm3 bone compartment and that the volume ratio of osteocyte cell body to bone compartment was 9.42% +/- 1.18%. Together, the average total length of the processes, the average surface area, and the average volume of one osteocyte were 1070 +/- 145 microm, 1509 +/- 113 microm2, and 394 +/- 49 microm3, respectively. It is possible to reconstruct the real architecture of the osteocyte network and obtain morphometric data from fluorescently labeled osteocytes in chick calvaria. PMID- 15820147 TI - Convergent loss of an anciently duplicated, functionally divergent RH2 opsin gene in the fugu and Tetraodon pufferfish lineages. AB - We describe the complete opsin gene families from the sequenced fugu and Tetraodon pufferfish genomes. We report the convergent loss of function of an anciently duplicated, functionally divergent RH2 or "green-sensitive" opsin gene in both pufferfish lineages, designated RH2-2. In fugu, RH2-2 apparently ceased to function very recently following a transposon-induced deletion that truncated the N-terminal 115 amino acids from the translated protein. Although a lack of frameshift or nonsense mutations in the fugu RH2-2 pseudogene suggests that the gene was lost very recently in this lineage, we were unable to detect any evidence of a selective sweep associated with the fixation of the truncated allele from population data. Interspecific comparison of the remaining fugu RH2-2 coding sequence paradoxically indicates that the gene was under strong purifying selection until the truncation occurred. PMID- 15820148 TI - Murine pulmonary infection with Listeria monocytogenes: differential susceptibility of BALB/c, C57BL/6 and DBA/2 mice. AB - Murine listeriosis is a paradigm to understand host pathogen interactions. Airway infections with Listeria monocytogenes, although representing a serious problem in early onset neonatal listeriosis, has not been investigated in detail in animal models so far. Here, the susceptibility of BALB/c, DBA/2 and C57BL/6 mice towards an intratracheal (i.t.) infection with virulent L. monocytogenes EGDe and the attenuated variant L. monocytogenes EGD hlyW491A(pERL3-CMVGFP) is reported. The course of infection was characterized by determination of bacterial numbers in the organs and assessment of the health condition of the mice. The distribution and cellular localization of Listeria in the airways was assessed by immunocytochemistry and confocal and electron microscopy. The differential susceptibility of inbred mouse strains to airway infections with L. monocytogenes could be assigned to the major virulence factor listeriolysin O. Resistant C57BL/6 mice were not affected by the two listerial strains. In contrast, BALB/c and DBA/2 mice showed differential susceptibility towards L. monocytogenes EGDe and attenuated bacteria, with all the mice being killed by the wild-type bacteria but rarely by the variant that secretes a listeriolysin of only 10% activity of that of the wild-type toxin. Thus, listeriolysin is a decisive factor for differential susceptibility against Listeria. After i.t. application, bacteria were predominantly localized in the peribronchiolar space and invaded alveolar macrophages but rarely lung epithelial cells. Dissemination from the lung into the deep organs started almost immediately after application, although a pulmonary bacterial reservoir remained during the first 4 days. PMID- 15820149 TI - The role of MAPK signal pathways during Francisella tularensis LVS infection induced apoptosis in murine macrophages. AB - Francisella tularensis is a highly virulent intracellular pathogen responsible for tularemia. This bacterium is capable of infecting many mammalian species and various cell types, but little is known about the mechanisms of survival and interactions with host cells. We examined the number of infected host cells, cytotoxicity and the role of apoptosis or necrosis in infection-induced cell death. Our results demonstrate that F. tularensis LVS induces apoptosis of infected macrophages within 10 h. At later time points we were also able to detect a dramatic increase in the proportion of necrotic macrophages. We investigated the signalling pathways involved in infection-induced cell death by analysing three mitogen-activated protein kinase (MAPK) pathways that are known to be activated by LPS stimulation; p42/p44 MAPK (Erk1/2), transcription factor c Jun and p38 MAPK. We identified post-translational activation of both p42 MAPK and p44 MAPK by phosphorylation at threonine and tyrosine residues after infection. Furthermore, treatment of infected cells with MEK1/2 inhibitors abrogated phosphorylation of p42/p44 MAPK and inhibited macrophage apoptosis and necrosis after infection. In contrast, phosphorylation and kinase activity of p38 MAPK was significantly lower in F. tularensis-infected cells, and inhibition of p38 MAPK activity induced apoptosis in uninfected cells. When we monitored JNK dependent phosphorylation of the transcription factor c-Jun, we did not observe any reactivity with either SAPK/JNK or phospho-SAPK/JNK antibodies at any time point. In conclusion, we demonstrate that F. tularensis LVS infection induces macrophage apoptosis. This process requires activation of the p42/p44 MAPK pathway and is associated with reduced p38 MAPK activity, indicating that infection-induced cell death can be caused by perturbation of these two signalling pathways. PMID- 15820150 TI - Epidemiological impact of vaccination on the dynamics of two childhood diseases in rural Senegal. AB - Measles and pertussis are ubiquitous vaccine-preventable diseases, which remain an important public health problem in developing countries. Hence, developing a deep understanding of their transmission dynamics remains imperative. To achieve this, we compared the impact of vaccination at both individual and population levels in a Senegalese rural community. This study represents the first such comparative study in tropical conditions and constitutes a point of comparison with other studies of disease dynamics in developed countries. Changes in the transmission rates of infections are reflected in their mean ages at infection and basic reproductive ratio calculated before and after vaccination. We explored persistence of both infections in relation to population size in each village and found the inter-epidemic period for the whole area using wavelets analysis. As predicted by epidemiological theory, we observed an increase in the mean age at infection and a decrease in the reproductive ratio of both diseases. We showed for both the pre- vaccination and vaccine eras that persistence depends on population size. After vaccination, persistence decreased and the inter-epidemic period increased. The observed changes suggest that vaccination against measles and pertussis induced a drop in their transmission. Similarities in disease dynamics to those of temperate regions such as England and Wales were also observed. PMID- 15820151 TI - Silencing viruses by RNA interference. AB - Post-transcriptional gene silencing (PTGS) makes possible new approaches for studying the various steps of the viral cycle. Plus-strand RNA viruses appear to be attractive targets for small interfering RNAs (siRNAs), as their genome functions as both mRNA and replication template. PTGS creates an alternative to classic reverse genetics for viruses with either negative-strand or double stranded RNA genomes and for those with a large genome. PTGS allows modification of the expression of a given cellular gene as a means to elucidate its role in the viral cycle and in virus-host cell interactions, and to investigate cellular pathways involved in viral pathogenesis. It also allows the creation of new animal models of human diseases. In addition, PTGS already appears to be a promising new therapeutic tool to fight viral multiplication and dissemination through the host and to prevent inflammation and virus-induced pathogenesis, including virus-induced tumorigenesis. PMID- 15820152 TI - Efficacy of cell-free antigens in evaluating cell immunity and inducing protection in a murine model of histoplasmosis. AB - Histoplasma capsulatum is a dimorphic pathogenic fungus that causes a wide spectrum of disease when mycelial fragments are inhaled. Resistance to H. capsulatum is dependent on cellular immunity mediated by T cells and macrophages. Here we standardized the production of extracts containing cell-free antigens (CFAgs) and observed their efficacy in evaluating cellular immunity during murine histoplasmosis. CFAgs induced a more potent delayed-type hypersensitivity (DTH) response in H. capsulatum-infected mice than did histoplasmin-a classical antigen. This DTH response to CFAgs is able to determine the immune status of infected mice and to predict their death. Moreover, CFAgs stimulated spleen cells from immune mice to produce higher amounts of gamma interferon (IFN-gamma) in vitro. Finally, immunization with CFAgs protected against a lethal inoculum of H. capsulatum. These results demonstrate that CFAgs may be useful for the evaluation of cellular immune response and as a potential source for the development of a vaccine against histoplasmosis. PMID- 15820153 TI - In vivo hierarchy of immunodominant and subdominant HLA-A*0201-restricted T-cell epitopes of HBx antigen of hepatitis B virus. AB - A polyepitopic CD8+ T-cell response is critical for the control of hepatitis B virus (HBV) infection. The HBV X protein (HBx) is a multifunctional protein that is important for the viral life cycle and for host-virus interactions. The aim of this study was to analyze the immunogenicity and dominance of various HLA-A*0201 restricted HBx-derived epitopes. For this purpose, we immunized HLA-A*0201 transgenic mice with HBx-derived peptides and DNA. This is a powerful model for studying the induction of HLA-A*0201-restricted immune responses in vivo, as these mice possess a cytotoxic T lymphocyte (CTL) repertoire representative of HLA-A2.1 individuals. We used cytotoxic tests and enzyme-linked immunosorbent spot (ELISPOT) assays to study the induction of specific cytotoxic and interferon (IFN)-gamma-secreting T cells. This allowed us to classify the HBx epitopes according to their T-cell activation capacity. After endogenous processing of the antigen synthesized in vivo after DNA-based immunization, we found that the HBx specific T-cell response is targeted against one immunodominant epitope. Furthermore, following peptide immunization, we identified six additional novel subdominant T-cell epitopes. Inclusion of well-characterized epitopic sequences of HBx in a new vaccine for chronic HBV infections could help to broaden the T cell response. PMID- 15820154 TI - Quantitative appraisal of murine filariasis confirms host strain differences but reveals that BALB/c females are more susceptible than males to Litomosoides sigmodontis. AB - Litomosoides sigmodontis, a rodent filarial nematode, can infect inbred laboratory mice, with full development to patency in the BALB/c strain. Strains such as C57BL/6 are considered resistant, because although filarial development can occur, circulating microfilariae are never detected. This model system has, for the first time, allowed the power of murine immunology to be applied to fundamental questions regarding susceptibility to filarial nematode infection. As this is a relatively new model, many aspects of the biology remain to be discovered or more clearly defined. We undertook a major analysis of 85 experiments, to quantitatively assess differences in filarial survival and reproduction in male versus female and BALB/c versus C57BL/6 mice over the full course of infection. This large dataset provided hard statistical support for previous qualitative reviews, including observations that the resistant phenotype of C57BL/6 mice is detectable as early as 10 days postinfection (dpi). An unexpected finding, however, was that filarial survival was reduced in male BALB/c mice compared to their female counterparts. Worm recovery as well as the prevalence and density of microfilariae were higher in female compared with male BALB/c mice. Therefore, L. sigmodontis bucks the filarial trend of increased susceptibility in males. This could be partially explained by the different anatomical locations of adult L. sigmodontis versus lymphatic filariae. Interestingly, the effects of BALB/c sex upon microfilaremia were independent of worm number. In summary, this study has significantly refined our understanding of the host-L. sigmodontis relationship and, critically, has challenged the dogma that males are more susceptible to filarial infection. PMID- 15820155 TI - Microarray analysis reveals expression regulation of Wnt antagonists in differentiating osteoblasts. AB - Wnt signaling has been implicated in regulating bone formation by controlling osteoblast proliferation and function. Although stabilization of beta-catenin by Wnt has been shown to increase alkaline phosphatase expression and osteoblast differentiation, the precise role of Wnt signaling during the process of osteoblast differentiation is largely unknown. In this study, we used microarray technology to investigate expression regulation of Wnt signaling components during in vitro osteoblast differentiation. Expression was analyzed during bone morphogenetic protein 2 (BMP2)-induced osteoblast differentiation of murine C2C12 and MC3T3 cells and data were compared with expression in BMP2-treated NIH3T3 fibroblasts. During osteoblast differentiation, particularly strong expression regulation of the Wnt antagonists Sfrp2 (secreted frizzled related protein 2) and Wif1 (Wnt inhibitory factor 1) was observed in the late phase of differentiation. In situ expression analysis in murine tail vertebrae supported Wif1 expression during late phase bone cell differentiation, since Wif1 was found to be expressed in vivo in trabecular, but not in cortical bone. We further analyzed the effects of continuous activation of Wnt signaling by lithium chloride and observed that osteoblast differentiation was reduced, as measured by expression of osteoblast marker genes encoding alkaline phosphatase, osteocalcin, and osterix, as well as by the amount of calcium release. Taken together, our data indicate that endogenous expression of Wnt antagonists by osteoblasts provides a negative Wnt feedback loop which is essential in controlling osteoblast maturation. PMID- 15820156 TI - Which parameters on magnetic resonance imaging determine Q waves on the electrocardiogram? AB - Studies have demonstrated that patients with Q-wave infarctions on the electrocardiogram (ECG) frequently have nontransmural scar formation, whereas non Q-wave infarctions may have transmural scars. The precise pathophysiologic substrate that underlies Q waves remains unclear. Magnetic resonance imaging (MRI) is the preferred technique to evaluate patients who have infarction because information can be obtained on function, contractile reserve (viability), and scar tissue. Consecutive patients (n = 69) who had coronary artery disease and a history of myocardial infarction underwent MRI; the protocol included MRI at rest, small-dose dobutamine MRI, and contrast-enhanced MRI. Parameters included left ventricular ejection fraction, left ventricular volumes, end-diastolic wall thickness and contractile reserve in the infarct region, transmurality and spatial extent of scar tissue, total scar score, and quantified percent left ventricular scar tissue. MRI data were related to the presence/absence of Q waves on the ECG. Q waves were present in 39 patients (57%). Univariate analysis identified transmurality, spatial extent, total scar score, and quantified percent scar tissue as predictors of Q waves. Multivariate analysis demonstrated that quantified percent scar tissue was the single best predictor of Q waves on the ECG. A cut-off value of 17% infarcted tissue of the left ventricle yielded a sensitivity and specificity of 90% to predict the presence/absence of Q waves. When quantified percent scar tissue was removed from the model, spatial extent of infarction was the best predictor. Thus, Q waves on the ECG correlate best with quantified percent scar tissue on contrast-enhanced MR images. PMID- 15820157 TI - Comparison of the usefulness of Doppler-derived deceleration time versus plasma brain natriuretic peptide to predict left ventricular remodeling after mechanical revascularization in patients with ST-elevation acute myocardial infarction and left ventricular systolic dysfunction. AB - The correlation between Doppler deceleration time (DT) and brain natriuretic peptide (BNP) and their predictive value for detecting left ventricular (LV) remodeling in patients who are treated with primary percutaneous intervention for infarction and LV dysfunction are unknown. Fifty-six patients (64 +/- 12 years of age; 11 women) who had a first ST-segment elevation myocardial infarction and systolic dysfunction that was successfully treated with direct primary coronary intervention underwent 2-dimensional Doppler echocardiographic and plasma BNP evaluation at days 1 and 3 and 1 and 6 months after the index infarction. Repeat coronary angiograms were obtained at 1 and 6 months. Because of previous consistent evidence, 3 days after the index infarction was the time point of comparison between BNP and DT values. Echocardiographic LV remodeling was defined as an increase in end-diastolic volume index above baseline values of 2 x SD. Ventricular remodeling occurred in 20 patients (36%). Multivariate analyses that included BNP level, Doppler DT, echocardiographic measurements of systolic function, peak creatine kinase, and anterior infarct location showed Doppler DT to be the only predictor of LV remodeling (odds ratio 0.963, 95% confidence interval 0.936 to 0.990, p = 0.008). The optimal cutoff for DT in the prediction of 6-month LV remodeling was <136 ms (sensitivity 75%, specificity 97%, accuracy 81%, area under receiver-operating characteristic curve 0.90). Thus, in patients who have a first ST-segment elevation myocardial infarction and LV systolic dysfunction that is successfully treated with primary percutaneous coronary intervention, Doppler-derived DT 3 days after index infarction is more effective than BNP level in detecting patients who are at higher risk for 6-month LV remodeling. PMID- 15820158 TI - Usefulness of electroanatomical mapping to differentiate between right ventricular outflow tract tachycardia and arrhythmogenic right ventricular dysplasia. AB - Differentiation between the different right ventricular rhythm disorders and specifically between arrhythmogenic right ventricular dysplasia (ARVD) and right ventricular outflow tract (RVOT) tachycardias has important clinical implications but remains a clinical challenge. We tested the hypothesis that the spatial association of local electrographic parameters may be used to discriminate between these 2 entities. Electroanatomic mapping of the right ventricle was performed in 3 groups: patients who had typical RVOT tachycardia, a control group of patients who had no ventricular arrhythmias, and patients who had a diagnosis of ARVD based on clinical, electrocardiographic, and structural findings. Electroanatomic mapping in the RVOT tachycardia group showed normal electrographic parameters throughout the right ventricle (unipolar electrographic amplitude 9.9 +/- 0.9 mV, duration 55 +/- 1 ms, amplitude/duration 0.193 +/- 0.022) that were no different from those in the control group. In contrast, dysplastic regions in the ARVD group were characterized by significantly lower amplitude (unipolar 3.6 +/- 0.4 mV), prolonged electrographic duration (unipolar 73 +/- 4 ms), and a decreased amplitude/duration ratio (unipolar 0.054 +/- 0.008) compared with the unaffected zones in the same hearts and with all regions in the RVOT and control groups. Thus, endocardial electrographic parameters do not differ between patients who have RVOT and control patients. RVOT tachycardia can be differentiated from ARVD by the absence of abnormal right ventricular electrographic findings. This ability may have important clinical implications and supports the concept of different underlying mechanisms for these 2 entities. PMID- 15820159 TI - Usefulness of Doppler assessment of pulmonary vein and left atrial appendage flow following pulmonary vein isolation of chronic atrial fibrillation in predicting recovery of left atrial function. AB - Atrial fibrillation (AF) is a widespread condition that causes significant morbidity and mortality. Recently, pulmonary venous (PV) isolation using radiofrequency ablation has been used successfully to exclude the pulmonary venous ostia, resulting in correction of AF. Further, miniaturized high-frequency ultrasound phased-array transducers currently provide Doppler and 2-dimensional imaging during the ablation procedure. We examined atrial function and its determinants using intracardiac echocardiography before and after PV isolation in 45 patients who had chronic AF (56 +/- 11 years old). PV, left atrial (LA) appendage, and mitral and tricuspid flows were recorded. Recovery of booster pump function (defined by the presence of mitral inflow A wave, LA appendage a-wave, and PV A-reversal wave velocities >10 cm/s) was observed in 39 of 45 patients (86.6%). PV flow systolic wave before and after ablation correlated with the degree of LA booster pump function after PV isolation. An early systolic PV flow peak velocity >57.47 cm/s predicted "good" LA booster pump function recovery with 96% specificity. Diastolic LA appendage emptying in AF correlated (p <0.001) and predicted good LA booster pump function with 92% specificity for velocities >46.4 cm/s. Thus, monitoring LA function during PV isolation for chronic AF is feasible. Most patients recovered LA booster pump function immediately after PV isolation, and the degree of recovery correlated with LA reservoir function. Preserved reservoir function during AF is predictive of satisfactory recovery of booster pump function after PV isolation. PMID- 15820161 TI - Effect of gender on treatment, resource utilization, and outcomes in congestive heart failure in Quebec, Canada. AB - This study assessed whether a gender gap exists in therapy, procedure use, and outcomes in patients who have a new diagnosis of congestive heart failure (CHF) at the population level. The Quebec hospital discharge database was linked with the physician and drug claims databases to identify a cohort of patients who had a diagnosis of CHF (code 428 in the International Classification of Diseases, Ninth Revision) between January 1998 and December 2002. There were 16,017 men and 16,622 women. Women who had CHF were older (78 +/- 11 vs 73 +/- 11 years, p <0.001), had more hypertension (41% vs 28%, p <0.001) and hyperlipidemia (18% vs 14%, p <0.001) but less frequent myocardial infarction (19% vs 25%, p <0.001). Women were less likely to see a cardiologist (30% vs 34%, p <0.001) and required a longer hospital stay (12.0 +/- 14.8 vs 10.6 +/- 13.3 days, p <0.001). During the first year after a first CHF hospitalization, women were less like to have an assessment of left ventricular function (61% vs 65%, p <0.001), diagnostic cardiac catheterization (11% vs 15%, p <0.001), and revascularization procedure (4% vs 6%, p <0.001). Women were less likely to be prescribed an angiotensin converting enzyme inhibitor (60% vs 66%) and more likely to be prescribed a beta blocker (38% vs 34%). Women and men had similar yearly numbers of rehospitalizations for CHF (1.4 +/- 1.0 vs 1.5 +/- 1.0) and emergency room visits (1.7 +/- 1.2 vs 1.8 +/- 1.3). The adjusted risk of death was minimally higher in men than in women (hazard ratio 1.06, 95% confidence interval 1.03 to 1.1, p <0.01). Thus, despite less frequent cardiologist assessment, fewer cardiac related procedures, and less frequent use of standard medical therapy, clinical outcomes in women and men who had CHF were similar. PMID- 15820160 TI - The N-terminal Pro-BNP investigation of dyspnea in the emergency department (PRIDE) study. AB - The utility of aminoterminal pro-brain natriuretic peptide (NT-proBNP) testing in the emergency department to rule out acute congestive heart failure (CHF) and the optimal cutpoints for this use are not established. We conducted a prospective study of 600 patients who presented in the emergency department with dyspnea. The clinical diagnosis of acute CHF was determined by study physicians who were blinded to NT-proBNP results. The primary end point was a comparison of NT-proBNP results with the clinical assessment of the managing physician for identifying acute CHF. The median NT-proBNP level among 209 patients (35%) who had acute CHF was 4,054 versus 131 pg/ml among 390 patients (65%) who did not (p <0.001). NT proBNP at cutpoints of >450 pg/ml for patients <50 years of age and >900 pg/ml for patients >or=50 years of age were highly sensitive and specific for the diagnosis of acute CHF (p <0.001). An NT-proBNP level <300 pg/ml was optimal for ruling out acute CHF, with a negative predictive value of 99%. Increased NT proBNP was the strongest independent predictor of a final diagnosis of acute CHF (odds ratio 44, 95% confidence interval 21.0 to 91.0, p <0.0001). NT-proBNP testing alone was superior to clinical judgment alone for diagnosing acute CHF (p = 0.006); NT-proBNP plus clinical judgment was superior to NT-proBNP or clinical judgment alone. NT-proBNP measurement is a valuable addition to standard clinical assessment for the identification and exclusion of acute CHF in the emergency department setting. PMID- 15820162 TI - Is operative closure of a small ventricular septal defect required after an episode of infective endocarditis? PMID- 15820163 TI - Cardiovascular benefits of nuts. PMID- 15820164 TI - Minimizing right ventricular pacing. PMID- 15820165 TI - Effect of both elevated troponin-I and peripheral white blood cell count on prognosis in patients with suspected myocardial injury. AB - We found a high white blood cell count (>11,000/mul) to be of additive prognostic value to high troponin-I levels in predicting risk of recurrent nonfatal myocardial infarctions and all-cause mortality in patients who present with acute coronary syndromes and non-ST-elevation myocardial infarctions. A high troponin-I level or white blood cell count increased the odds ratio of an event to 2.2 (95% confidence interval 1.0 to 4.73, p = 0.05), but high values for the 2 markers increased the odds ratio to 4.5 (95% confidence interval 1.42 to 14.21, p = 0.01). PMID- 15820166 TI - Compliance as a critical consideration in patients who appear to be resistant to aspirin after healing of myocardial infarction. AB - The hypothesis that aspirin resistance is often due to noncompliance was investigated. One hundred ninety patients with a history of myocardial infarction were evaluated using arachidonic acid-stimulated light aggregometry at 3 different time points: while receiving their usual daily aspirin, after not receiving aspirin for 7 days, and 2 hours after the observed ingestion of aspirin 325 mg. At the first time point, 17 patients (9%) failed to show aspirin inhibition of platelet aggregation, but 2 hours after observed aspirin ingestion, aspirin inhibition was observed in all but 1 patient. PMID- 15820167 TI - Safety and effectiveness of transdermal nicotine patch in smokers admitted with acute coronary syndromes. AB - An analysis of smokers admitted with acute coronary syndrome who received transdermal nicotine therapy and those who did not was performed. Propensity analysis was used to match patients. Transdermal nicotine therapy appears safe and does not have an effect on the mortality of patients with acute coronary syndromes. PMID- 15820168 TI - Relation of the use of lipid-lowering medications prior to percutaneous coronary intervention to the incidence of intraprocedural adverse angiographic events. AB - The use of lipid-lowering medications at the time of percutaneouscoronary intervention (PCI) has been shown to have a favorable effect on rates of cardiac enzyme elevation and major adverse cardiac events (MACEs), but the effect of these medications on angiographically identifiable intraprocedural coronary events during PCI has not previously been investigated. A retrospective review of 81 patients was performed and demonstrated that the use of lipid-lowering medications at the time of PCI was associated with a reduced incidence of angiographically identifiable intraprocedural events (odds ratio 0.13, 95% confidence interval 0.04 to 0.40). A multivariate analysis revealed that lipid lowering medications and hyperlipidemia did not predict MACEs independent of the occurrence of these angiographic events. PMID- 15820169 TI - Relation of myocardial perfusion to mortality after primary percutaneous coronary intervention. AB - In patients who undergo primary percutaneous coronary intervention (PCI), poor post-PCI myocardial blush is associated with increased mortality, even when epicardial perfusion is adequate. This observation has important implications for the methods of evaluating primary PCI results and the strategies used to improve myocardial reperfusion. PMID- 15820170 TI - Frequency of myopathy in patients receiving lovastatin. AB - Lovastatin (Mevacor) 20 mg is being considered for nonprescription availability. Because the most severe untoward consequence of therapy with any statin is rhabdomyolysis, the clinical data for lovastatin pertaining to this adverse event were reviewed. Evidence to date, based on almost 2 decades of experience, points to an extremely low risk for myopathy and rhabdomyolysis associated with lovastatin. PMID- 15820171 TI - Predicting coronary heart disease risk using multiple lipid measures. AB - Principal component analysis was used to summarize variations among 7 lipid measures included in the Framingham Offspring Study (n = 2,694). An overall measure combining information from the 7 lipids was compared with conventional lipid measures in adjusted survival analyses and was found to be a superior predictor of coronary heart disease risk. PMID- 15820172 TI - Feasibility, safety, and morphologic predictors of outcome of repeat percutaneous balloon mitral commissurotomy. AB - Early and mid-term results of repeat percutaneous balloon mitral commissurotomy (PBMC) were analyzed in 35 patients with symptomatic valvular restenosis: 12 patients (34%) after first successful PBMC and 23 patients (66%) after successful surgical closed mitral commissurotomy. Twenty-one patients had bilateral fused commissures, and 14 patients had unilateral or bilateral split commissures. Mitral valve area gain was significantly greater in the group with fused commissures compared with the group with split commissures (0.6 +/- 0.2 vs 0.3 +/ 0.2 cm(2), respectively, p = 0.04). PMID- 15820173 TI - Long-term follow-up of patients with obstructive hypertrophic cardiomyopathy treated with dual-chamber pacing. AB - In this study, patients with obstructive hypertrophic cardiomyopathy (HC) were treated with dual-chamber pacemaker therapy. Long-term follow-up analysis showed that dual-chamber pacemaker therapy in selected patients resulted in a significant reduction in symptoms and in the left ventricular outflow tract gradient, which was maintained up to 10 years after implantation. Dual-chamber pacing is of potential long-term benefit in selected groups of patients with obstructive HC. PMID- 15820174 TI - Effect on right ventricular volume of percutaneous Amplatzer closure of atrial septal defect in adults. AB - In adult patients with atrial septal defects (ASDs), right ventricular (RV) cavity size may return to normal after operative closure. This study demonstrated improved RV volumes and right atrial areas in 20 adult patients after successful transcatheter closure of large ASDs. RV volumes decreased by 22%, 30%, and 41% at 1 day, 1 month, and 6 months, respectively, after the procedure. Right atrial areas decreased by 5%, 23%, and 26%, respectively, over the same time. PMID- 15820175 TI - Comparison of aortic stiffness in patients with juvenile forms of ascending aortic dilatation with versus without Marfan's syndrome. AB - Aortic stiffness in 78 patients with juvenile forms of ascending aortic dilation with Marfan's syndrome was compared with aortic stiffness in 17 patients without Marfan's syndrome. In the ascending aorta and aortic arch, aortic stiffness was similarly increased in the 2 groups. In the descending aorta, however, aortic stiffness was increased only in patients with Marfan's syndrome. PMID- 15820176 TI - Relation between actual heart rate and autonomic effects of beta blockade in healthy men. AB - This study evaluated the effect of beta blockade on cardiac autonomic control and its dependence on heart rate change. The relations between RR interval duration, heart rate variability (HRV), and baroreflex sensitivity (BRS) were studied in 111 healthy men and in 21 male volunteers before and after a 100-mg oral dose of metoprolol. HRV and BRS correlated significantly with mean RR (r = 0.39, r = 0.57). Metoprolol administration increased both mean RR (from 996 to 1,176 ms, p <0.001), BRS from 24 to 36 ms/mm Hg (p = 0.003), and the SD of RR from 61 to 74 ms (p = 0.05). However, metoprolol-induced changes of HRV and BRS became insignificant (p = 0.69 and 0.48) after they were normalized to the same cycle length, suggesting that the improvement of cardiac autonomic control after beta blockade could be explained by a change of heart rate. PMID- 15820177 TI - Effect of exercise training on VO2peak and left ventricular systolic function in recent cardiac transplant recipients. AB - This study examined the effect of 12 weeks of combined aerobic and resistance training on aerobic capacity, left ventricular ejection fraction, and arterial afterload during submaximal cycle exercise in 18 recent (<1 month from transplantation) cardiac transplant recipients. The main finding of this study is that 12 weeks of combined aerobic and resistance training is an effective intervention to increase aerobic capacity in this population. However, exercise training was not associated with favorable improvements in left ventricular systolic function, because stroke volume and ejection fraction were reduced after training because of an increase in arterial afterload. PMID- 15820178 TI - Relation of decreased ankle-brachial index to prevalence of atherosclerotic risk factors, coronary artery disease, aortic valve calcium, and mitral annular calcium. AB - Coronary artery disease was present in 89 of 118 patients (75%) with a decreased ankle-brachial index (ABI) and in 34 of 118 age- and gender-matched patients (29%) with a normal ABI (p <0.001). Aortic valve calcium or mitral annular calcium was present in 81 of 118 patients (69%) with a decreased ABI and in 43 of 118 patients (36%) with a normal ABI (p <0.001). PMID- 15820179 TI - Effect of light-to-moderate alcohol consumption on age-associated arterial stiffening. AB - Increased thickness and stiffness of large arteries may contribute to why aging is the most important risk for cardiovascular diseases. Arterial stiffness, intimal medial thickness, and alcohol intake were measured in 563 subjects. A U shaped relation was found between alcohol intake and a stiffness index, with the lowest index in moderate drinkers, which may partially explain the relation between alcohol and cardiovascular disease. PMID- 15820180 TI - Comparison of tissue Doppler dynamics to Doppler flow in evaluating left atrial appendage function by transesophageal echocardiography. AB - Tissue Doppler imaging (TDI) patterns from the left atrial appendage (LAA) were evaluated by transesophageal echocardiography. Reproducible, characteristic triphasic or biphasic tissue velocities similar to Doppler flow of the LAA were obtained. Patient peak TDI velocities correlated well with flow and were measurable in atrial fibrillation. Patients with an embolic cerebrovascular accident and in sinus rhythm had higher tissue TDI velocities from the LAA compared with patients without an event, and the groups had similar flow velocities. Hence, Doppler tissue contraction dynamics determined by TDI may complement flow velocities in evaluating LAA function for risk assessment of thromboembolism. PMID- 15820181 TI - Usefulness of left atrial emptying fraction to predict exercise capacity in patients with normal systolic left ventricular function and without myocardial ischemia. AB - The association of left atrial volume at rest and the left atrial emptying fraction with exercise capacity during stress echocardiography was examined in patients with normal left ventricular function and without ischemia. The left atrial emptying fraction, along with body mass index, was found to be an independent predictor of poor exercise capacity (<5 METs). PMID- 15820182 TI - Comparison of tissue Doppler and propagation velocity to invasive measures for measuring left ventricular filling pressures. AB - The utility of tissue Doppler and propagation velocity in identifying patients with abnormal left ventricular filling pressures was assessed in 50 patients who underwent catheterization and echocardiography on the same day. The ratios of the peak velocity of early mitral inflow to early mitral annular velocity (E/Ea) and the velocity of propagation (E/Vp) were compared with invasive measurements of pre-A-wave left ventricular pressures. Echocardiography measures were 77% to 92% accurate in identifying patients with a pre-A-wave pressure >15 mm Hg. Tissue Doppler imaging is more accurate than propagation velocity. PMID- 15820183 TI - Effects of age, gender, and left ventricular mass on septal mitral annulus velocity (E') and the ratio of transmitral early peak velocity to E' (E/E'). AB - The ratio of the transmitral early peak velocity (E) evaluated by conventional Doppler imaging over early diastolic mitral annulus velocity (E') evaluated by tissue Doppler imaging has been proposed as a noninvasive marker for left ventricular filling pressure. We evaluated 174 normal patients and 86 patients with hypertension and LV hypertrophy to assess the effects of age, gender, and left ventricular hypertrophy on E' and E/E'. Age appeared to be the strongest determinant of E' and E/E', suggesting that in normal patients and in those with left ventricular hypertrophy, age-dependent cut-off values should be considered. PMID- 15820184 TI - What about statistical confounding? PMID- 15820185 TI - Fibrillation cycle length and fibrillatory rate obtained from peak frequency of fibrillation waves. PMID- 15820186 TI - Aging and biomedicine 2005: where should we go from here? PMID- 15820187 TI - Aging hearts and vessels: masters of adaptation and survival. PMID- 15820188 TI - Aging-associated changes in cardiac gene expression. AB - Cardiovascular diseases (e.g., vascular diseases, strokes, heart failure) reach epidemic proportions in the elderly and are the primary limits to survival in man. Age-associated changes in heart structure and function represent the major risk factors in heart failure (HF) syndromes and are associated with altered patterns of gene expression that can generally be seen as relative changes in the abundance of gene transcripts. An understanding of the molecular mechanisms underlying these changes should be tantamount to defining a genetic basis for aging; however, the analysis of processes as complicated as aging requires an accounting of biological diversity. Until recently, most of the changes in transcript abundance were identified one at a time, but the advent of gene expression arrays has permitted rapid, large-scale expression profiling. This has provided information about the dynamics of total gene expression, which can be used to identify pathways and elucidate regulatory events that may be affected during senescence or in response to disease. Importantly, very large sample sizes or meta-analyses of studies of smaller sample sizes should be sufficient to account for the diversity of altered gene expression that directs alterations in specific molecular pathways, which underlie changes in cardiac structure and function in senescence and disease. PMID- 15820189 TI - Gene expression profiling studies of aging in cardiac and skeletal muscles. AB - To examine transcriptional alterations associated with aging in skeletal muscle and the heart, we and others have used DNA microarrays to compare the gene expression profile of young and old animals. Aging results in a differential gene expression pattern specific to each tissue, and most alterations can be completely or partially prevented by caloric restriction (CR) in both heart and skeletal muscle. Transcriptional patterns of tissues from calorie-restricted animals suggests that CR retards the aging process by reducing endogenous damage and by inducing metabolic shifts associated with specific transcriptional profiles. These studies demonstrate that DNA microarrays can be used in cardiovascular aging research to generate panels of hundreds of transcriptional biomarkers, providing a new tool to measure biological age of cardiac and skeletal muscles and to test interventions designed to retard aging in these tissues. PMID- 15820190 TI - Aging, telomeres, and atherosclerosis. AB - Although the level and pace of population aging display high geographical variability, virtually all countries have been experiencing growth in their elderly population, particularly in developed nations. Because aging is a major risk factor for atherosclerosis and associated disease, it is of up most importance to unravel the molecular mechanisms involved in vascular aging. Telomeres are specialized DNA-protein structures located at the ends of eukaryotic chromosomes whose length is progressively reduced in most somatic cells during aging. It is accepted that telomere exhaustion contributes to organismal ageing at least by impairing cell proliferation and viability. An emerging question is whether telomere erosion contributes to atherosclerosis. Here we discuss recent advances on the molecular control of telomere length in vascular cells, as well as animal and human studies that address the role of telomeres in vascular pathobiology. Although the interrelationships between telomere length and cardiovascular disease appear obvious, a chief question that remains unanswered is whether telomere ablation is cause of vascular injury or a surrogate phenomenon. PMID- 15820191 TI - Mitochondrial function and myocardial aging. A critical analysis of the role of permeability transition. AB - Mitochondria have been suggested to be causally linked to age-related alterations through respiratory chain dysfunction and formation of reactive oxygen species, leading to damage of mitochondrial DNA. Impaired biosynthesis of respiratory chain and ATP synthase subunits encoded by mitochondrial genes would set up a vicious cycle contributing to the aging process. Mitochondria are also involved in the increased susceptibility to ischemic injury observed in aged hearts, a process where the mitochondrial permeability transition pore (PTP) may play a role. Here, we analyze (i) the possible mechanisms through which PTP opening might contribute to age-related myocardial alterations; (ii) the available evidence of an increased probability of PTP opening in mitochondria isolated from aged tissues; (iii) the current methodological limitations that complicate the elucidation of causal relationships between PTP opening, mitochondrial dysfunction, and myocardial aging. PMID- 15820192 TI - Protection in the aged heart: preventing the heart-break of old age? AB - The aged heart has a diminished functional and adaptive reserve capacity, an increased susceptibility to incur damage (e.g., as a result of ischemia), and a limited practical ability for repair/regeneration. Thus, there has been considerable interest to harness the heart's endogenous capacity to resist such damage, known as ischemic preconditioning (IPC), as well as other cardioprotective mechanisms. However, the translation of basic research findings into clinical practice has largely been inadequate because there have been few if any successful implementations in terms of viable therapies activating cardioprotection mechanisms to limit infarct size. Here, we provide an overview of the general mechanisms of cardioprotection, changes in the structure and function of the aged heart, and the current knowledge regarding cardioprotection in aged heart. The problems and opportunities for successful bench-to-bedside translation of cardioprotection in the elderly are discussed. PMID- 15820193 TI - Adenosine-mediated cardioprotection in the aging myocardium. AB - With aging, it appears the heart's ability to withstand injury declines markedly. Unfortunately, the incidence of ischemic disorders increases dramatically with age. Though the genesis of the ischemia-intolerant phenotype is incompletely understood (and likely multi-factorial), it may involve changes in intrinsic cardioprotective responses. In this respect we and others have interrogated the role of the adenosine receptor (AR) system in dictating ischemic tolerance and the impact of age on AR-mediated cardioprotection. It is intriguing to note ARs impact on many processes implicated in myocardial 'aging': adenosine counters Ca2+ influx and oxidant injury, modifies substrate metabolism to improve tolerance, is pro-angiogenic, inhibits myocardial fibrosis, and can limit apoptosis. Thus, dysregulation of the AR system could contribute to many features of aged hearts (including ischemic intolerance). The latter is borne out by observations that AR-mediated protective responses decline with intrinsic tolerance and that transgenic manipulation of the AR system restores intrinsic tolerance and protective responses in aged hearts. Mechanisms underlying failure in adenosinergic protection remain undefined. Here we review data on the effects of aging on cardiovascular AR transcription and expression, generation of signal (adenosine formation), and protective signaling coupled to ARs. PMID- 15820194 TI - Sympathetic neuronal regulation of the heart in aging and heart failure. AB - The documentation of preferential activation of the cardiac sympathetic outflow in patients with heart failure swept aside an entrenched notion that there was a functional sympathetic denervation of the failing heart and provided a theoretical basis for the clinical evaluation of beta-adrenergic blocker therapy in this condition. The demonstration that heightened sympathetic nervous system activity is central to the pathogenesis and progression of congestive heart failure (CHF) has now led to the rational use of beta-adrenoceptor blockade in CHF. More recently, it has also emerged that the aging heart exhibits some of the characteristic changes in autonomic control which are seen in CHF. Accordingly, alterations in cardiac sympathetic nerve function are now thought to contribute also to the pathophysiology of the aging heart. Furthermore, there is evidence that in humans, sympathoexcitatory rostral projections of brainstem noradrenergic neurons to the forebrain are important in the sympathetic nervous activation of both heart failure and aging. Given these similarities, in this review we compare and contrast the neurobiology of the sympathetic nervous system in the failing heart and the healthy, aging heart, and consider whether the sympathetic activation accompanying aging may, perhaps, underlie and contribute to the neural pathophysiology of heart failure. Our conclusion is, on balance, that this proposition is not supported by the available evidence. PMID- 15820195 TI - Inflammatory markers and cardiovascular health in older adults. AB - In the past decade inflammatory markers have emerged as strong independent risk indicators for cardiovascular disease. Even though adults over the age of 65 experience a high proportion of such events, most epidemiologic data are from middle-aged populations. In this review we examine the role that inflammatory markers play in the prediction of incident cardiovascular disease specifically in older adults. In studies of adults < 65 years, IL-6, TNFalpha and IL-10 levels have been shown to predict cardiovascular outcomes. The data on C-reactive protein are inconsistent, but CRP levels appear to be less useful in old-age than in middle-age. Fibrinogen levels predict mortality but in a non-specific manner. In the elderly inflammatory markers are non-specific measures of health and predict both disability and mortality even in the absence of clinical cardiovascular disease. Thus it is possible that, in older age-groups, interventions designed to prevent cardiovascular disease through the modulation of inflammation would also be helpful in reducing disability and mortality. PMID- 15820196 TI - Aging and plasminogen activator inhibitor-1 (PAI-1) regulation: implication in the pathogenesis of thrombotic disorders in the elderly. AB - Thrombotic cardiovascular diseases increase in incidence in the elderly, a tendency dependent on the age-related changes in vascular and hemostatic systems that include platelets, coagulation, and fibrinolytic factors as well as in the endothelium. The hypercoagulability of and advanced sclerotic changes in the vascular wall may contribute to the increased incidence of thrombosis in the elderly. One of the important key genes for aging-associated thrombosis is plasminogen activator inhibitor-1 (PAI-1), a principal inhibitor of fibrinolysis. The expression of PAI-1 is not only elevated in the elderly but also significantly induced in a variety of pathologies associated with the process of aging. These conditions include obesity, insulin resistance, emotional stress, immune responses, and vascular sclerosis/remodeling. Several cytokines and hormones, including tumor necrosis factor-alpha, transforming growth factor-beta, angiotensin II, and insulin, positively regulate the gene expression of PAI-1. The recent epidemic in obesity with aging in the industrialized society may heighten the risk for thrombotic cardiovascular disease because adipose tissue is a primary source of PAI-1 and cytokines. Emotional or psychosocial stress and inflammation also cause the elevated expression of PAI-1 in an age-specific pattern. Thus, PAI-1 could play a key role in the progression of cardiovascular aging by promoting thrombosis and vascular (athero)sclerosis. Further studies on the genetic mechanism of aging-associated PAI-1 induction will be necessary to define the basis for cardiovascular aging in relation to thrombosis. PMID- 15820197 TI - Endothelial aging. AB - Aging is considered to be the major risk factor for the development of atherosclerosis and, therefore, for coronary artery disease. Apart from age associated remodeling of the vascular wall, which includes luminal enlargement, intimal and medial thickening, and increased vascular stiffness, endothelial function declines with age. This is most obvious from the attenuation of endothelium-dependent dilator responses, which is a consequence of the alteration in the expression and/or activity of the endothelial NO synthase, upregulation of the inducible NO synthase, and increased formation of reactive oxygen species. Aging is also associated with a reduction in the regenerative capacity of the endothelium and endothelial senescence, which is characterized by an increased rate of endothelial cell apoptosis. PMID- 15820198 TI - Vascular consequences of menopause and hormone therapy: importance of timing of treatment and type of estrogen. AB - Premenopausal women have a lower risk for cardiovascular events, and mortality due to coronary vascular disease (CVD) in premenopausal women is rare. These facts suggest that endogenous estrogens, such as estradiol, protect the cardiovascular system, and several observational studies and a few small clinical studies conducted in healthy and younger postmenopausal women support this hypothesis. In contrast, two large randomized clinical trials (RCTs), using conjugated equine estrogens and conducted in older women with established CVD or without overt CVD, failed to demonstrate protection against CVD by exogenous estrogens. These divergent findings have resulted in confusion with regard to the association between estrogen deficiency and CVD in postmenopausal women. In order to reconcile these contradictory findings, it is necessary to examine the pathophysiology associated with age-dependent changes within the vessel wall and to compare the pharmacology of different types of estrogens. Understanding age dependent changes in vascular pathology and the pharmacology of different estrogens may facilitate the development of therapeutic strategies for hormone replacement therapy (HRT) that would be effective in delaying vascular remodeling leading to CVD following menopause. In this review we provide an overview of the impact of menopause and estrogen deficiency on vascular remodeling and emphasize the importance of timing and type of estrogen to achieve maximum benefits with regard to reducing the risk of CVD. PMID- 15820199 TI - Evolution and modulation of age-related medial elastocalcinosis: impact on large artery stiffness and isolated systolic hypertension. AB - Arteriosclerosis, characterized by remodeling and stiffening of large elastic arteries is the most significant manifestation of vascular aging. The increased stiffening is believed to originate from a gradual mechanical senescence of the elastic network, alterations in cross-linking of extracellular matrix components, fibrosis and calcification of elastic fibers (medial elastocalcinosis). The stiffening of large arteries reduces their capacitance and accelerates pulse wave velocity, thus contributing to a widening of pulse pressure and to the increased prevalence of isolated systolic hypertension with age. Current antihypertensive drugs were mainly designed to reduce peripheral resistance and are not adequate to alter the pathological process of vascular stiffening or even to selectively reduce systolic blood pressure in isolated systolic hypertension. This review puts forward the concept that elastocalcinosis is a valuable therapeutic target and presents evidence that this process can be prevented and reversed pharmacologically. PMID- 15820200 TI - In vivo gene transfer of parvalbumin improves diastolic function in aged rat hearts. AB - OBJECTIVE: Diastolic dysfunction is a characteristic finding of the aged mammalian heart. Parvalbumin acts as a Ca2+ sink and enhances relaxation in skeletal muscle, and overexpression of parvalbumin in myocardium increased cardiac relaxation in vitro as well as in vivo. Therefore, the objective of this study is to test the hypothesis that in vivo gene transfer of parvalbumin will improve diastolic dysfunction in aged rat heart. METHODS: We used adenovirus to transfer parvalbumin into two different rat models of aging: the Fischer 344 (F344) and the Fischer 344 x Brown Norway F1 hybrid (F344 x BN). Cardiac function was measured and compared after gene transfer. RESULTS: In vivo overexpression of parvalbumin in both rat aging models had no effect on systolic parameters but reduced left ventricular diastolic pressure and the time course of pressure decline. Overexpression of parvalbumin also improved the force frequency relationship in senescent rats. CONCLUSION: In vivo overexpression of parvalbumin improves diastolic dysfunction in two rat models of senescence, and this effect is independent of the rat strain investigated. The results show promise that gene therapy of parvalbumin may address the impaired Ca2+ homeostasis and diastolic dysfunction without an increase in energy expenditure. PMID- 15820201 TI - Smooth muscle cells deficient in osteopontin have enhanced susceptibility to calcification in vitro. AB - OBJECTIVE: Vascular calcification is an actively regulated process, correlating with cardiovascular morbidity and mortality especially in patients with diabetes and chronic renal diseases. Osteopontin (OPN) is abundantly expressed in human calcified arteries and inhibits vascular calcification in vitro and in vivo. How OPN functions in vascular calcification, however, is less clear. METHODS: Smooth muscle cells (SMCs) were isolated from aortas of OPN knock-out (OPN-/-) and wild type (OPN+/+) mice. RESULTS: OPN-/- SMCs were identical to OPN+/+ SMCs in morphology and stained positively for SM lineage proteins, desmin, smooth muscle alpha-actin and SM22alpha. No spontaneous calcification was observed in OPN-/- SMCs under normal culture conditions or in medium containing 1%, 3%, or 5% fetal bovine serum. However, when cultured in medium containing elevated concentrations of inorganic phosphate, an inducer of vascular calcification, a significantly higher calcification was observed in OPN-/- SMCs compared to OPN+/+ SMCs that, in response to elevated phosphate, synthesized and secreted OPN into the culture. Finally, retroviral transduction of mouse OPN cDNA into OPN-/- SMCs rescued the calcification phenotype of the cells. CONCLUSION: These results are the first to demonstrate an inhibitory role of endogenously produced OPN on SMC calcification, suggesting a novel feedback mechanism where OPN produced locally by the SMCs may serve as an important inducible inhibitor of vascular calcification. PMID- 15820202 TI - Cardiac-specific effects of parathyroid hormone-related peptide: modification by aging and hypertension. AB - OBJECTIVE: Parathyroid hormone-related peptide (PTHrP) improves heart function of post-ischemic and stunned myocardium and is released from the heart under ischemic conditions. Hypertrophic hearts from spontaneously hypertensive rats (SHR) develop a reduced ischemic tolerance, show reduced expression of PTHrP and develop paradoxical effects in regard to PTHrP. We hypothesized that PTHrP is causally involved in reduced ischemic tolerance of hypertrophied hearts. This hypothesis was tested by addition of a cardiac-specific PTHrP agonist or antagonist during ischemia and investigation of the functional recovery during the early phase of reperfusion. METHODS: Hearts from male normotensive adult (6 months) or old (12 months) Wistar and SHR rats were perfused at a constant flow for 20 min and then exposed for 30 or 15 min to global zero-flow ischemia followed by 30 min reperfusion. PTHrP agonist (PTHrP1-36) or antagonist (5Ile,23Trp,36Tyr-PTHrP1-36) (each 100 nmol/l) were added briefly before the onset of ischemia to ensure that they were present at the beginning of reperfusion. Heart function was determined by insertion of a balloon catheter into the left ventricle. Left ventricular developed pressure (LVDP), dP/dtmax, dP/dtmin, left ventricular end-diastolic pressure (LVeDP), heart rate (HR) and coronary resistance (CR) were recorded. RESULTS: Reduced post-ischemic recovery in old SHR was confirmed. Hearts from all four groups responded normally to exogenous PTHrP with a positive chronotropic effect under non-ischemic conditions. In hearts from adult normotensive rats, a beneficial effect of released endogenous PTHrP was confirmed. However, addition of the cardiac specific PTHrP antagonist during ischemia significantly improved post-ischemic recovery in hearts from old normotensive rats and SHR. This beneficial effect of the antagonist was accompanied by a significant reduction in post-ischemic LVeDP and was more pronounced in adult SHR. This effect was also observed when the hearts were paced (4 Hz). Upon short-term ischemia (15 min), in the absence of ischemia-induced rigor contraction, the antagonist improved LVDP recovery in hearts from old normotensive rats. CONCLUSION: In summary, a protective effect of released endogenous PTHrP was confirmed for hearts from adult normotensive rats. This effect is converted into an opposite effect in hearts from SHR and old normotensive rats. Therefore, released endogenous PTHrP can contribute to reduced ischemic tolerance in hypertrophied hearts and during aging. PMID- 15820203 TI - Renovascular effects of sympathetic cotransmitters ATP and NPY are age-dependent in spontaneously hypertensive rats. AB - OBJECTIVE: Hypertension is characterized by sympathetic overactivity. Neuropeptide Y (NPY) and ATP are cotransmitters of norepinephrine (NE) and regulate renovascular resistance. The present study analyzes sympathetic nonadrenergic neurotransmission in hypertensive (SH-SP) and normotensive (WKY) rats. In addition, adult and young hypertensive rats were compared to investigate the role of aging on sympathetic nonadrenergic cotransmission in hypertensive disease. METHODS: Pressor responses to renal nerve stimulations (RNS) and drugs were measured on isolated perfused kidneys of young (8-10 weeks) and adult (18-24 weeks) WKY, and SH-SP rats. RESULTS: RNS evoked contractions at 1 Hz were resistant to blockade by the alpha-adrenoceptor antagonist phentolamine (1 microM) but abolished by the P2 receptor blocker suramin (100 microM). Compared to adult WKY, RNS-induced pressor responses were unchanged in adult SH-SP and young WKY, but significantly greater in young SH-SP rats. The NPY-Y1 receptor antagonist BIBP3226 (1 microM) reduced phentolamine-resistant pressor responses in adult and young WKY, young SH-SP, but not in adult SH-SP rats. In contrast to WKY and young SH-SP rats, exogenously perfused NPY (0.1 microM) was unable to potentiate RNS-induced, phentolamine-resistant pressor responses in adult SH-SP rats. NE and the stable ATP analogue alpha,beta-mATP increased the perfusion pressor response more potently in adult SH-SP than in WKY rats. CONCLUSIONS: Neuronally released NPY plays a major role in potentiating RNS-induced nonadrenergic pressor responses in kidneys of WKY and young SH-SP rats. In adult SH-SP rats NPY fails to enhance these responses. In this hypertensive model ageing seems to be associated with a loss of a modulatory role of renal NPY Y1 receptors. Since pressor responses to NE and ATP are higher in SH-SP animals, functional NPY-Y1 receptor downregulation might be an adaptive mechanism. PMID- 15820204 TI - Age-associated changes in electrophysiologic remodeling: a potential contributor to initiation of atrial fibrillation. AB - OBJECTIVE: Although the incidence of atrial fibrillation (AF) increases with age, the cellular electrophysiological changes that render the atria of aged individuals more susceptible to AF remain poorly understood. We hypothesized that dispersion of atrial repolarization increases with aging, creating a substrate for initiation of AF. METHODS: Four groups of dogs were studied: adult and old dogs in normal sinus rhythm (SR) and adult and old dogs with chronic AF (CAF) induced by rapid atrial pacing. In each dog, action potentials (AP) were recorded with microelectrodes from isolated endocardial preparations of four regions of right atrium and three regions of left atrium. Two indices of AP duration (APD) heterogeneity were obtained in each dog by calculating standard deviation (SD) and the coefficient of variation (COV=[SD/mean] x 100%). RESULTS: In SR groups, APD averaged across all regions was significantly longer in old than in adult tissues. Both indices of APD heterogeneity were higher in old dogs in comparison to adult. At both ages, CAF was associated with significant APD shortening and a decrease in APD adaptation to rate. While CAF significantly increased both indices of APD heterogeneity in adult dogs, it significantly decreased them in old dogs. CONCLUSIONS: The increase of spatial variability in repolarization in old atria may contribute to the initiation of AF in the aged. CAF-induced APD shortening and a decrease in APD adaptation appear to be important for the maintenance of sustained AF in both adult and old atria. The CAF-induced increase in dispersion of repolarization may be important for AF stabilization in adults, while previously reported fibrosis and slowed conduction of premature beats may be important in the old for both AF initiation during SR and subsequent stabilization of AF. PMID- 15820205 TI - Heart function and molecular biological parameters are comparable in young adult and aged rats after chronic myocardial infarction. AB - OBJECTIVE: To test the hypothesis that IL-1beta and IL-6 play a pivotal role after myocardial infarction (MI) particularly in aged rats. METHODS: Chronic MI was induced in young adult (3.5 months) and aged (18 months) female Sprague Dawley rats by ligation of the left coronary artery. Sham-operated animals of corresponding age served as controls. Heart function was measured by catheterization 4 weeks after MI. The expression of IL-1beta, IL-6, TGF-beta isoforms, ANF, and components of the extracellular matrix (pro-collagen I and III, colligin, MMP-2 and TIMP2) was measured by ribonuclease protection assay. RESULTS: Aged control rats differed from young adult rats in that LV-developed pressure (LVDP) was higher (161 vs. 147 mmHg, p<0.05) in response to the elevated total peripheral resistance (0.71 vs. 0.47 mmHg ml min/kg, p<0.05). Contractility was reduced in aged controls as indicated by decreased LV dP/dt (8.106 vs. 10.606 mmHg/s, p<0.05). LV function was severely depressed in both MI groups (reduction in LVDP by about 35% and LV dP/dt by about 30%, increase in LVEDP to 24 mmHg) while RVP and RV dP/dt markedly increased by about 100%. This was not different between both MI groups. ANF expression as a marker of hypertrophy was induced in both MI groups, but less pronounced in the LV of aged rats. Also, the mRNA expression pattern was qualitatively comparable, but showed gradual differences. CONCLUSION: These results indicate that aged rats compensate well for hemodynamic overload induced by MI. Also, the mechanisms of myocardial post-MI remodeling are comparable in young adult and aged rats. PMID- 15820206 TI - Aging decreases vasoconstrictor responses of coronary resistance arterioles through endothelium-dependent mechanisms. AB - OBJECTIVE: Aging decreases coronary blood flow and maximal reserve capacity. Impaired blood flow capacity may be related to an increased vasoconstrictor capacity of coronary resistance vessels. This study tested the hypothesis that aging increases the vasoconstrictor responsiveness of coronary arterioles isolated from myocardium of young (4 months) and old (24 months) Fischer 344 rats. METHODS: Isolated coronary arterioles were cannulated and pressurized (60 cm H2O) via hydrostatic pressure reservoirs. RESULTS: Contrary to our hypothesis, aging decreased responsiveness of coronary arterioles to endothelin (ET, 1 x 10( 11)-3 x 10(-8) M), potassium chloride (KCl, 10-100 mM), and pressure-induced myogenic responses (0-140 cm H2O). Removal of the endothelium from coronary arterioles increased vasoconstriction to all agonists; however, age-related KCl vasoconstrictor response differences remained, suggesting that K+ channel activity and/or the relative contribution of specific K+ channels to maintenance of vascular smooth muscle membrane potential may change with age. Removal of the endothelium, in addition to increasing responsiveness, eliminated aging-induced differences in ET- and pressure-induced vasoconstriction. L-NAME (10(-5)) incubation resulted in a greater enhancement of spontaneous tone in arterioles from old rats compared to those of young rats. ETB (BQ-788, 3 x 10(-8)) receptor blockade eliminated the age-associated differences. CONCLUSION: Collectively, these data suggest an age-associated increase in endothelial modulation of coronary resistance vessel constriction. This enhanced endothelial attenuation of coronary arteriolar constriction appears to result from increased basal release of nitric oxide. These alterations of coronary vascular reactivity may contribute to age-induced redistribution of coronary blood flow and diminished cardiac function. PMID- 15820207 TI - Exaggerated coronary vasoreactivity to endothelin-1 in aged rats: role of protein kinase C. AB - OBJECTIVE: The interaction between advanced age and increased susceptibility to ischemic insult is well documented. Age-related increases in coronary vascular resistance, in part due to impaired dilator responses, have been reported. Our aim was to determine the role of endothelin-1 (ET-1) on enhanced constrictor responses in aged coronary arteries (CAs) and whether protein kinase C (PKC) signaling mechanisms impact ET-1 responses. METHODS: Vasoreactivity was assessed in CAs isolated from aged (24 months; n=16) and adult (4 months; n=21) male F344 rats following ET-1 (10(-10)-10(-8)) with and without specific ETA/ETB receptor antagonists (BQ-123, 1 microM; BQ-788, 30 nM) or the PKC inhibitor bisindolylmaleimide (Bis; 10(-6) M). Constrictor responses to KCl (80 mM) were also measured and voltage-gated Ca2+ channel (VGCC) determined in isolated coronary smooth muscle cells. Dilator responses to acetylcholine (ACH) and sodium nitroprusside (SNP) were assessed. RESULTS: Passive diameter was greater (357+/ 19 vs. 309+/-9; p<0.02) while spontaneous tone was similar in 24 months vs. 4 months. ET-1 resulted in greater constriction in 24 months vs. 4 months (79% vs. 67%; p<0.01). Group differences persisted following selective ETB inhibition with BQ-788 (p<0.02), while BQ-123 abolished contractile responses to ET-1. Importantly, inhibition of ET-1 constriction by Bis occurred in 24 months but not 4 months (p<0.01). Constrictor responses to KCl and peak VGCC current density were similar in 24 months vs. 4 months (48% vs. 50%). No age-related differences were observed in ACH- or SNP-mediated dilation. Western blotting revealed increases in Ca2+-sensitive PKCalpha, -betaI, and -betaII levels with age, while eNOS and ETA receptor protein levels were unchanged. CONCLUSION: Aberrant ETA constrictor responses and directional changes in PKC are likely to contribute to coronary vascular pathology with advanced age. PMID- 15820208 TI - The effects of aging and exercise training on endothelin-1 vasoconstrictor responses in rat skeletal muscle arterioles. AB - OBJECTIVES: The incidence of cardiovascular disease increases with advancing age. Vascular dysfunction has been linked to cardiovascular disease and aging, although most research has focused on endothelium-dependent vasodilator dysfunction. Another possible mechanism for this vascular dysfunction with aging is enhanced vasoconstrictor responsiveness of the resistance vasculature, and in particular, reactivity of arterioles to endothelin-1 (ET-1). We hypothesized that vasoreactivity to ET-1 would be greater in skeletal muscle arterioles from old rats, and that endurance exercise training would abolish differences in ET-1 responsiveness between young and old animals. METHODS: Young sedentary (YS; 4 months; n=18), old sedentary (OS; 24 months; n=17), young trained (YT; n=9) and old trained (OT; n=7) male Fischer 344 rats were used. Training modality was treadmill exercise at 15 m/min up a 15 degrees incline, 1 h/day, 5 days/week, for 12 weeks. Soleus and white gastrocnemius muscle first-order arterioles were isolated for in vitro experimentation. Intraluminal diameter was measured in response to increasing concentrations of ET-1 (10(-11) to 10(-8) M) or KCl (10 100 mM) in arterioles with an intact or denuded endothelium and with and without an ETA (BQ-123 [10(-6) M]) or ETB (BQ-788 [10(-8) M]) receptor antagonist present. RESULTS: There was an age-associated increase in gastrocnemius vasoconstrictor responsiveness and sensitivity to ET-1 in arterioles with intact endothelium (ET-1 EC50: YS, 5.2 E(-9)+/-1.1 E(-9) M; OS, 2.0 E(-9)+/-0.8 E(-9) M); neither removal of the endothelium nor ETB blockade abolished this difference in arteriolar sensitivity to ET-1 between old and young rats. In contrast, ETA inhibition abolished the greater sensitivity (EC50) of arterioles from old animals (ET-1 EC50: YS, 10 E(-9)+/-0.7 E(-9) M; OS, 10 E(-9)+/-1.5 E(-9) M). Gastrocnemius muscle arterioles exhibited an age-related reduction in KCl-induced vasoconstriction which was abolished with the removal of the endothelium. Soleus muscle arteriolar responses to ET-1 and KCl were unaffected by aging. Additionally, exercise training had no effect on ET-1 vasoconstriction of soleus or gastrocnemius muscle arterioles. CONCLUSIONS: Aging results in an augmented gastrocnemius muscle arteriolar vasoconstriction to ET-1 which is mediated through an enhanced ETA receptor signaling pathway and not through an ETB receptor mechanism associated with either the endothelium or vascular smooth muscle. These findings suggest that enhanced vascular ET-1 sensitivity in fast twitch skeletal muscles may play a role in the vascular dysfunction that is often associated with old age. PMID- 15820209 TI - MMP-9 regulates both positively and negatively collagen gel contraction: a nonproteolytic function of MMP-9. AB - OBJECTIVE: Constrictive remodeling accounts for lumen loss in postangioplasty restenosis. Matrix metalloproteinase-9 (MMP-9) has been shown to prevent constrictive remodeling in vivo. To investigate potential mechanisms for this observation, we investigated the role of MMP-9 in smooth muscle cell (SMC) mediated collagen gel contraction, an in vitro model of constrictive remodeling. METHODS: Fischer rat SMCs were stably transfected with a construct-expressing rat MMP-9 under the control of a tetracycline (Tet)-off promoter. SMCs were seeded in type I collagen gels (2.4 mg/ml) in the presence or not of tetracycline (1 microg/ml), and gel contraction was defined as the percentage of retraction of the collagen gel. The depletion of MMP-9 was obtained by using siRNA targeting MMP-9 mRNA or a blocking antibody. RESULTS: Gel contraction was significantly reduced at all times when MMP-9 was overexpressed (Tet-) as compared with the control condition (Tet+). However, MMP-9 depletion of control (Tet+) SMCs (using siRNA or antibody) also inhibited gel contraction. To resolve the apparent discrepancy and determine if MMP-9 exerts a dose-dependent biphasic effect on gel contraction, conditioned medium and purified rat-MMP-9 were prepared. Gel contraction was significantly increased by addition of 0.8 ng/ml of MMP-9, while high concentrations of MMP-9 (> or =100 ng/ml) inhibited contraction. The addition of BB94 and TIMP-1 did not alter the inhibitory or stimulatory effect of MMP-9. CONCLUSIONS: Our data suggest that MMP-9, independent of its proteolytic function, has a biphasic effect on SMC-mediated collagen gel contraction. Understanding the different roles of MMP-9 should allow the development of better therapeutic strategies for restenotic vascular disease. PMID- 15820211 TI - The antioxidant effect of tannic acid on the in vitro copper-mediated formation of free radicals. AB - Tannic acid (TA) has well-described antimutagenic and antioxidant activities. The antioxidant activity of TA has been previously attributed to its capacity to form a complex with iron ions, interfering with the Fenton reaction [Biochim. Biophys. Acta 1472, 1999, 142]. In this work, we observed that TA inhibits, in the micromolar range, in vitro Cu(II) plus ascorbate-mediated hydroxyl radical (*OH) formation (determined as 2-deoxyribose degradation) and oxygen uptake, as well as copper-mediated ascorbate oxidation and ascorbate radical formation (quantified in EPR studies). The effect of TA against 2-deoxyribose degradation was three orders of magnitude higher than classic *OH scavengers, but was similar to several other metal chelators. Moreover, the inhibitory effectiveness of TA, by the four techniques used herein, was inversely proportional to the Cu(II) concentration in the media. These results and the observation of copper-induced changes in the UV spectra of TA are indications that the antioxidant activity of TA relates to its copper chelating ability. Thus, copper ions complexed to TA are less capable of inducing ascorbate oxidation, inhibiting the sequence of reactions that lead to 2-deoxyribose degradation. On the other hand, the efficiency of TA against 2-deoxyribose degradation declined considerably with increasing concentrations of the *OH detector molecule, 2-deoxyribose, suggesting that the copper-TA complex also possesses an *OH trapping activity. PMID- 15820210 TI - Age-dependent changes in myocardial matrix metalloproteinase/tissue inhibitor of metalloproteinase profiles and fibroblast function. AB - OBJECTIVE: To evaluate the effects of aging on left ventricular (LV) geometry, collagen levels, matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) abundance, and myocardial fibroblast function. METHODS: Young (3-month-old; n=28), middle-aged (MA; 15-month-old; n=17), and old (23 month-old; n=16) CB6F1 mice of both sexes were used in this study. Echocardiographic parameters were measured; collagen, MMP, and TIMP levels were determined for both the soluble and insoluble protein fractions; and fibroblast function was evaluated. RESULTS: LV end-diastolic dimensions and wall thickness increased in both MA and old mice, accompanied by increased soluble protein and decreased insoluble collagen. Immunoblotting revealed differential MMP/TIMP profiles. Compared to MA levels, MMP-3, MMP-8, MMP-9, MMP-12, and MMP-14 increased, and TIMP-3 and TIMP-4 decreased in the insoluble fraction of old mice, suggesting increased extracellular matrix (ECM) degradative capacity. Fibroblast proliferation was blunted with age. CONCLUSION: This study, for the first time, identified specific differences in cellular and extracellular processes that likely contribute to age-dependent ECM remodeling. PMID- 15820212 TI - Identification of a novel zebrafish SULT1 cytosolic sulfotransferase: cloning, expression, characterization, and developmental expression study. AB - By searching the zebrafish expressed sequence tag database, we had identified two partial cDNA clones encoding the 5'- and 3'-regions of a putative cytosolic sulfotransferase (SULT). Using the reverse transcription-polymerase chain reaction (RT-PCR) technique, a full-length cDNA encoding this zebrafish SULT was amplified, cloned, and sequenced. Analysis of the sequence data revealed that this novel zebrafish SULT displays 49, 46, and 45% amino acid sequence identity to human SULT1A1, mouse SULT1D1, and rat SULT1C1. This zebrafish SULT therefore appears to belong to the SULT1 cytosolic SULT gene family. Recombinant zebrafish SULT (designated SULT1 isoform 4), expressed using the pGEX-2TK prokaryotic expression vector and purified from transformed Escherichia coli cells, migrated as a 35kDa protein upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among the endogenous compounds tested as substrates, the purified SULT1 isoform 4 displayed significant sulfating activities toward thyroid hormones, estrone, and dehydroepiandrosterone. The enzyme also showed activities toward a number of xenobiotics including some flavonoids, isoflavonoids, and other phenolic compounds, with a pH optimum at 7.0. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 28 and 37 degrees C. Among 10 divalent metal cations tested, Fe2+, Hg2+, Co2+, Zn2+, Cu2+, and Cd2+ exhibited dramatic inhibitory effects on the activity of the enzyme. Developmental expression study using RT PCR revealed that the zebrafish SULT1 isoform 4 showed a low level of expression in the segmentation period during the embryonic development, which gradually increased to a high level of expression throughout the larval stage onto maturity. PMID- 15820213 TI - A three-component dicamba O-demethylase from Pseudomonas maltophilia, strain DI 6: purification and characterization. AB - Dicamba O-demethylase is a multicomponent enzyme that catalyzes the conversion of the herbicide 2-methoxy-3,6-dichlorobenzoic acid (dicamba) to 3,6 dichlorosalicylic acid (DCSA). The three components of the enzyme were purified and characterized. Oxygenase(DIC) is a homotrimer (alpha)3 with a subunit molecular mass of approximately 40 kDa. FerredoxinDIC and reductaseDIC are monomers with molecular weights of approximately 14 and 45 kDa, respectively. EPR spectroscopic analysis suggested the presence of a single [2Fe-2S](2+/1+) cluster in ferredoxinDIC and a single Rieske [2Fe-2S](2+; 1+) cluster within oxygenaseDIC. Consistent with the presence of a Rieske iron-sulfur cluster, oxygenaseDIC displayed a high reduction potential of E(m,7.0) = -21 mV whereas ferredoxinDIC exhibited a reduction potential of approximately E(m,7.0) = -171 mV. Optimal oxygenaseDIC activity in vitro depended on the addition of Fe2+. The identification of formaldehyde and DCSA as reaction products demonstrated that dicamba O-demethylase acts as a monooxygenase. Taken together, these data suggest that oxygenaseDIC is an important new member of the Rieske non-heme iron family of oxygenases. PMID- 15820214 TI - The requirement for the hydrophobic motif phosphorylation of Ypk1 in yeast differs depending on the downstream events, including endocytosis, cell growth, and resistance to a sphingolipid biosynthesis inhibitor, ISP-1. AB - ISP-1 inhibits de novo sphingolipid biosynthesis and induces growth defects in both mammals and yeast (Saccharomyces cerevisiae). In our previous study, YPK1/SLI2 was identified as one of multicopy suppressor genes for ISP-1 in yeast. Ypk1 is proposed to be a downstream serine/threonine kinase of the sphingolipid signaling pathway in yeast. Other than resistance against ISP-1, Ypk1 is involved in at least two downstream events, namely cell growth and endocytosis. In this study, the effect of mutants of Ypk1 on these three downstream events was investigated. Among Ypk1 mutants, no 'kinase-dead' mutants complemented the defects in any of these three downstream events in the ypk1 null strain. One of the hydrophobic motif phosphorylation-deficient mutants of Ypk1, Ypk1(T662A) had the moderate kinase activity compared with the wild-type Ypk1. Ypk1(T662A) and the wild-type Ypk1 completely restored the slow-growth phenotype and fluid-phase endocytosis defect of the ypk1 null strain. However, unlike the wild-type Ypk1, Ypk1(T662A) lost the ability for the recovery of the ISP-1 resistance in the ypk1 null strain. Furthermore, the expression of Ypk1(T662A) in the wild-type strain showed a dominant-negative effect on the ISP-1-resistance activity. On the other hand, the cell growth revertant of the ypk1 null strain still showed the hypersensitive phenotype to ISP-1. These data suggest that the ISP-1-resistance pathway is under the regulation of the hydrophobic motif phosphorylation and is separated from the other pathways downstream of Ypk1. PMID- 15820215 TI - Ca2+- and voltage-dependent inactivation of the expressed L-type Ca(v)1.2 calcium channel. AB - Ca2+-dependent regulation of the ion current through the alpha1Cbeta2aalpha2delta 1 (L-type) calcium channel transiently expressed in HEK 293 cells was investigated using whole cell patch clamp method. Ca2+ or Na+ ions were used as a charge carrier. Intracellular Ca2+ was either buffered by 10 mM EGTA or 200 microM Ca2+ was added into non-buffered intracellular solution. Free intracellular Ca2+ inactivated permanently about 80% of the L-type calcium current. The L-type calcium channel inactivated during a depolarizing pulse with two time constants, tau(fast) and tau(slow). Free intracellular calcium accelerated both time constants. Effect on the tau(slow) was more pronounced. About 80% of the channel inactivation during brief depolarizing pulse could be attributed to a Ca2+-dependent mechanism and 20% to a voltage-dependent mechanism. When Na+ ions were used as a charge carrier, the L-type current still inactivated with two time constants that were 10 times slower and were virtually voltage-independent. Ca2+ ions stabilized the inactivated state of the channel in a concentration-dependent manner. PMID- 15820216 TI - Effect of age, vitamin D, and calcium on the regulation of rat intestinal epithelial calcium channels. AB - Transepithelial transport of calcium involves uptake at the apical membrane, movement across the cell, and extrusion at the basolateral membrane. Active vitamin D metabolites regulate the latter two processes by induction of calbindin D and the plasma membrane ATPase (calcium pump), respectively. The expression of calbindin D and the calcium pump declines with age in parallel with transepithelial calcium transport. The apical uptake of calcium is thought to be mediated by the recently cloned calcium channels-CaT1 (or ECaC2, TRPV6) and CaT2 (or ECaC1, TRPV5). The purpose of these studies was to determine whether there were age-related changes in intestinal calcium channel regulation and to identify the dietary factors responsible for their regulation. Young (2 months) and adult (12 months) rats were fed either a high calcium or low calcium diet for 4 weeks. The low calcium diet significantly increased duodenal CaT1 and CaT2 mRNA levels in both age groups, but the levels in the adult were less than half that of the young. The changes in calcium channel expression with age and diet were significantly correlated with duodenal calcium transport and with calbindin D levels. To elucidate the relative roles of serum 1,25(OH)2D3 and calcium in the regulation of calcium channel expression, young rats were fed diets containing varying amounts of calcium and vitamin D. Dietary vitamin D or exogenous 1,25(OH)2D3 more than doubled CaT1 mRNA levels, and this regulation was independent of dietary or serum calcium. These findings suggest that the apical calcium channels, along with calbindin and the calcium pump, may play a role in intestinal calcium transport and its modulation by age, dietary calcium, and 1,25(OH)2D3. PMID- 15820217 TI - Evaluation of spin trapping agents and trapping conditions for detection of cell generated reactive oxygen species. AB - Electron paramagnetic resonance with spin trapping is a useful technique to detect reactive oxygen species, such as superoxide radical anion (O2*-), a key species in many biological processes. We evaluated the abilities of four spin traps in trapping cell-generated O2*-: 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), 2-diethoxyphosphoryl-2-phenethyl-3,4-dihydro-2H-pyrrole N-oxide (DEPPEPO), 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO), and 5,5 dimethyl-1-pyrroline N-oxide (DMPO). Optimal experimental conditions for obtaining maximal signal intensity of O2*- adduct in a cellular system were first studied. The maximal intensities of BMPO, DEPMPO, and DMPO adducts were similar while DEPPEPO did not trap cell-generated O2*- induced by 1,6-benzo[a]pyrene quinone in a human mammary epithelial cell line (MCF-10A). BMPO and DEPMPO adducts were more stable, considering the stability of their maximal signal, than DMPO adduct in the tested cellular systems. In addition, we observed that O2*- spin adducts were reduced to their corresponding hydroxyl adducts in the cellular system. The selection of optimal spin trap in trapping cell-generated O2*- is discussed. PMID- 15820218 TI - Four promoters direct expression of the calpastatin gene. AB - Calpastatin is a specific endogenous protein inhibitor of the ubiquitous calcium dependent proteinases mu- and m-calpain. The calpain-calpastatin system is involved in various physiological and pathological processes. In the present study, we determined the bovine calpastatin gene structure and demonstrated that four promoters direct its expression. The gene harbours 35 exons spanning at least 130kb on genomic DNA. Its structure is similar to that of mouse, pig, and human gene. Transient transfection assays in both C2C12 and COS7 cell lines demonstrated that the putative promoter regions situated 5' to exon 1xa, 1xb, 1u, and 14t were functional. We also established that the region situated upstream exon 14t is subjected to a tissue specific regulation. The implication of numerous high-scoring cis acting transcriptional motifs which are present in these regions will need to be determined. The existence of four promoters suggests differential expression patterns which must have a physiological significance. PMID- 15820219 TI - Inhibition of erythrocyte acetylcholinesterase by n-butanol at high concentrations. AB - Erythrocyte acetylcholinesterase (AChE) is bound to the membrane by a complex glycosylphosphatidylinositol anchor, so the effect of alcohol on AChE activity may reflect direct and/or membrane-mediated effects. The indication of a direct interaction between n-butanol and AChE molecules is the activation/inhibition of AChE by occupation of the enzyme's active and/or regulatory sites by alcohol. The activation of AChE can occur only at low concentrations of alcohols, while at high concentrations AChE is inhibited. In this work the mechanism of inhibition of erythrocyte AChE by n-butanol at high concentrations was studied. The values of activity, calculated assuming parabolic competitive inhibition, which implies that one or two molecules of inhibitor bind to the enzyme, fit well to the experimental values. From the values of the inhibition constants it was concluded that at high n-butanol concentrations two alcohol molecules usually interact with AChE. PMID- 15820220 TI - Divergent folding pathways of two homologous proteins, BPTI and tick anticoagulant peptide: compartmentalization of folding intermediates and identification of kinetic traps. AB - Bovine pancreatic trypsin inhibitor (BPTI) and tick anticoagulant peptide (TAP) are two structurally homologous proteins, which have been shown to exhibit distinct mechanisms of oxidative folding. We demonstrate here differences of their folding properties using the technique of disulfide scrambling. Two extensively unfolded homologous isomers (beads-form) of BPTI (Cys5-Cys14, Cys30 Cys38, Cys51-Cys55) and TAP (Cys5-Cys15, Cys33-Cys39, Cys55-Cys59) were allowed to refold in parallel via disulfide shuffling of 13 potential isomers to form the native structure. Folding intermediates were trapped by acid quenching and analyzed by HPLC. The results reveal the following diversities: (a) there are two predominant folding intermediates of BPTI and seven well-populated folding intermediates of TAP. One of the two predominant BPTI intermediates (Cys5-Cys14, Cys30-Cys51, Cys38-Cys55) contains a native disulfide Cys30-Cys51 and constitutes about 34% of the total BPTI folding intermediates. In contrast, the TAP counterpart (Cys5-Cys15, Cys33-Cys55, Cys39-Cys59) represents only 5% of the total TAP intermediates; (b) three isomers of TAP sharing a stable non-native disulfide bond Cys15-Cys33 are shown to act as kinetic traps of TAP folding. Their counterparts are conspicuously absent in the BPTI folding; and (c) most significantly, folding intermediates of BPTI are found to be energetically compartmentalized, whereas most folding intermediates of TAP are inter convertible and exist in dynamic equilibrium. Our studies further demonstrate optimal concentrations of denaturant required for destabilization of kinetic traps and acceleration of TAP folding. PMID- 15820221 TI - Overexpression of manganese superoxide dismutase protects against ATP depletion mediated cell death of proximal tubule cells. AB - We have previously shown that in vivo renal ischemia/reperfusion results in ATP depletion, oxidant production, and manganese superoxide dismutase (MnSOD) inactivation. Current studies were designed to compare the effect of ATP depletion (Antimycin A treatment) on cell death pathways using renal proximal tubular cells and identical cells that overexpress MnSOD. ATP depletion in wild type cells induced an apoptotic cascade that involved caspase 9 activation; MnSOD overexpressing cells afforded protection against apoptosis. This protection did not appear to involve a cytochrome c-related mechanism, but may be related to altered levels of nitric oxide within the cell. Further studies suggested that nitric oxide was required to protect the renal cells from caspase-mediated cell death. Interestingly, treatment of renal cell extracts with reductants (DTT and ascorbate) enhanced caspase activation. Taken together, these results suggest that cysteine nitrosylation may be playing a role in caspase dysfunction in cells overexpressing MnSOD following ATP depletion. PMID- 15820222 TI - Proton transfer from exogenous donors in catalysis by human carbonic anhydrase II. AB - In the site-specific mutant of human carbonic anhydrase in which the proton shuttle His64 is replaced with alanine, H64A HCA II, catalysis can be activated in a saturable manner by the proton donor 4-methylimidazole (4-MI). From 1H NMR relaxivities, we found 4-MI bound as a second-shell ligand of the tetrahedrally coordinated cobalt in Co(II)-substituted H64A HCA II, with 4-MI located about 4.5 A from the metal. Binding constants of 4-MI to H64A HCA II were estimated from: (1) NMR relaxation of the protons of 4-MI by Co(II)-H64A HCA II, (2) the visible absorption spectrum of Co(II)-H64A HCA II in the presence of 4-MI, (3) the inhibition by 4-MI of the catalytic hydration of CO2, and (4) from the catalyzed exchange of 18O between CO2 and water. These experiments along with previously reported crystallographic and catalytic data help identify a range of distances at which proton transfer is efficient in carbonic anhydrase II. PMID- 15820223 TI - Energetics of carbohydrate binding to Momordica charantia (bitter gourd) lectin: an isothermal titration calorimetric study. AB - Physico-chemical and carbohydrate binding studies have been carried out on the Momordica charantia (bitter gourd) seed lectin (MCL). The lectin activity is maximal in the pH range 7.4-11.0, but decreases steeply below pH 7.0. The lectin activity is mostly unaffected in the temperature range 4-50 degrees C, but a sharp decrease is seen between 50 and 60 degrees C, which could be correlated to changes in the structure of the protein as seen by circular dichroism and fluorescence spectroscopy. Isothermal titration calorimetric studies show that the tetrameric MCL binds two sugar molecules and the binding constants (Kb), determined at 288.15 K, for various saccharides were found to vary between 7.3 x 10(3) and 1.52 x 10(4)M(-1). The binding reactions for all the saccharides investigated were essentially enthalpy driven, with the binding enthalpies (DeltaHb) at 288.15 K being in the range of -50.99 and -43.39 kJ mol(-1), whereas the contribution to the binding reaction from the entropy of binding was negative, with values of binding entropy (DeltaSb) ranging between -99.2 and 72.0 J mol(-1)K(-1) at 288.15 K. Changes in heat capacity (DeltaCp) for the binding of disaccharides, lactose and lactulose, were significantly larger in magnitude than those obtained for the monosaccharides, methyl-beta-D galactopyranoside, and methyl-alpha-D-galactopyranoside, and could be correlated reasonably well with the surface areas of these ligands. Enthalpy-entropy compensation was observed for all the sugars studied, suggesting that water structure plays an important role in the overall binding reaction. CD spectroscopy indicates that carbohydrate binding does not lead to significant changes in the secondary and tertiary structures of MCL, suggesting that the carbohydrate binding sites on this lectin are mostly preformed. PMID- 15820225 TI - Bipolar disorder and polymorphisms in the dysbindin gene (DTNBP1). AB - BACKGROUND: Several studies support the dysbindin (dystrobrevin binding protein 1) gene (DTNBP1) as a susceptibility gene for schizophrenia. We previously reported that variation at a specific 3-locus haplotype influences susceptibility to schizophrenia in a large United Kingdom (UK) Caucasian case-control sample. METHODS: Using similar methodology to our schizophrenia study, we have investigated this same 3-locus haplotype in a large, well-characterized bipolar sample (726 Caucasian UK DSM-IV bipolar I patients; 1407 ethnically matched controls). RESULTS: No significant differences were found in the distribution of the 3-locus haplotype in the full sample. Within the subset of bipolar I cases with predominantly psychotic episodes of mood disturbance (n = 133) we found nominally significant support for association at this haploptype (p < .042) and at SNP rs2619538 (p = .003), with a pattern of findings similar to that in our schizophrenia sample. This finding was not significant after correction for multiple testing. CONCLUSIONS: Our data suggest that variation at the polymorphisms examined does not make a major contribution to susceptibility to bipolar disorder in general. They are consistent with the possibility that DTNBP1 influences susceptibility to a subset of bipolar disorder cases with psychosis. However, our subset sample is small and the hypothesis requires testing in independent, adequately powered samples. PMID- 15820226 TI - Decreased hippocampal expression of the susceptibility gene PPP3CC and other calcineurin subunits in schizophrenia. AB - BACKGROUND: Calcineurin (CaN) is a phosphatase involved in synaptic plasticity. A haplotype of the PPP3CC gene, which encodes the gamma isoform of the catalytic subunit (CaN A), has been associated with schizophrenia. However, the distribution of CaN A gamma is not established, nor whether its expression changes in schizophrenia. METHODS: CaN A expression was analyzed in the hippocampal formation of 13 patients with schizophrenia and 12 controls. All three isoforms were examined, using in situ hybridization histochemistry, RT-PCR, and laser-assisted microdissection. CaN A protein was assessed using ELISA and immunohistochemistry. CaN A mRNAs were also measured in rats treated with haloperidol or chlorpromazine. RESULTS: CaN was prominent in excitatory neurons. CaN A alpha and A beta isoforms were abundant in all subfields, but CaN A gamma was not reliably detected in CA1. CaN A protein, and all three mRNAs, were decreased in schizophrenia. The mRNA reductions were present in all subfields measured, except CA1. CaN A mRNAs were unaltered in the antipsychotic-treated rats. CONCLUSIONS: Decreased CaN expression extends the evidence for aberrant hippocampal synaptic plasticity in schizophrenia, which particularly affects glutamatergic transmission, and which leaves CA1 relatively unaffected. Reduced expression of PPP3CC may underlie its genetic involvement in the disorder. PMID- 15820227 TI - Differential changes in apolipoprotein E in schizophrenia and bipolar I disorder. AB - BACKGROUND: This study extends an initial finding of increased levels of apoE in Brodmann's area (BA) 9 from subjects with schizophrenia to determine if apoE is altered in other brain regions and in brains from subjects with bipolar I disorder (BID). METHODS: ApoE was quantified apoE in BA 9, 10, 40, 46 and caudate putamen from control (n = 18), schizophrenic (n = 19) and BID (n = 8) subjects using Western blotting. RESULTS: In schizophrenia, there was increased apoE in BA9 (mean +/- SEM: schizophrenia 3.8 +/- .18 vs. control 3.2 +/- .19) and BA46 (schizophrenia 2.7 +/- .26 vs. control 1.6 +/- .20). In BID, increased levels of the apolipoprotein were detected in the caudate putamen (BID 3.3 +/- .44 vs. control 2.4 +/- .19) and BA9 (BID 4.0 +/- .27 vs. control 3.2 +/- .19) with a decrease in apoE being measured in BA10 (BID 1.6 +/- .16 vs. control 3.9 +/- .53). CONCLUSIONS: This study has shown disease specific, regionally discrete changes in levels of apoE in brain obtained post mortem from schizophrenic and BID subjects. Our data adds weight to the hypothesis that changes in the levels of apolipoproteins may be involved in the pathologies of schizophrenia and bipolar disorder. PMID- 15820228 TI - Alterations in cell adhesion molecule L1 and functionally related genes in major depression: a postmortem study. AB - BACKGROUND: Current research in depression aims to delineate genes involved in neuronal plasticity that are altered in the disease or its treatment. We have shown antidepressant induced increases in three interrelated genes, cell adhesion molecule L1 (CAM-L1), laminin, and cAMP response element binding protein (CREB), and a reciprocal decrease in these genes consequent to stress. Presently we hypothesized that CAM-L1, CREB, and laminin may be altered in post mortem brains of depressed subjects. METHODS: Studies were performed in the prefrontal and in the ventral parieto-occipital cortices, of 59 brains from depressed, bipolar, and schizophrenic subjects, and normal controls, obtained from the Stanley Foundation Brain Collection. mRNA and protein levels were determined by RT-PCR and Western blot analysis, respectively. RESULTS: Levels of CAM-L1 and of phosphorylated CREB (pCREB) were increased in the prefrontal cortex of the depressed group, while CAM L1, laminin and pCREB were decreased in the parieto-occipital cortex. Depressed subjects receiving antidepressants differed from subjects not receiving antidepressants in the expression of CAM-L1 and laminin in the parieto-occipital cortex, and in the expression of pCREB in the prefrontal cortex. CONCLUSIONS: The present findings of specific alterations in depression and antidepressant treatment particularly in CAM-L1 suggest that this gene may play an important role in the pathophysiology and treatment of depression. PMID- 15820229 TI - Specific motion processing pathway deficit during eye tracking in schizophrenia: a performance-matched functional magnetic resonance imaging study. AB - BACKGROUND: The neural mechanisms underlying smooth pursuit eye movement (SPEM) abnormalities in schizophrenia are not well understood. Previous evidence suggests that a deficit in the processing of internal representations of object motion (extraretinal motion) contributes to SPEM deficits in patients. Functional magnetic resonance imaging (fMRI) activation was compared between patients and control subjects to determine whether schizophrenia patients exhibit abnormal cerebral activation in regions associated with extraretinal motion processing during SPEM. METHODS: Patients and control subjects were selected based on matched performance in the closed-loop gain. Despite similar performance on closed-loop pursuit gain, patients showed consistent deficits in extraretinal motion based on predictive pursuit. In the magnet, subjects were tested using a traditional smooth-pursuit task that elicits closed-loop response. RESULTS: Patients had reduced pursuit-related activation in several known extraretinal motion processing areas including frontal and supplemental eye fields, medial superior temporal cortex, and anterior cingulate. Patients also showed increased activation in medial occipitotemporal cortex. CONCLUSIONS: These results provide functional anatomic evidence supporting reduced function in the extraretinal motion processing pathway in schizophrenia. Increased activation in medial occipitotemporal cortex suggests an increased dependence on immediate retinal motion information, which may be used to compensate for reduced extraretinal signaling during sustained visual tracking. PMID- 15820230 TI - Fronto-thalamo-striatal gray and white matter volumes and anisotropy of their connections in bipolar spectrum illnesses. AB - BACKGROUND: Neurons in the basal ganglia are connected to areas of prefrontal cerebral cortex involved in higher cognitive functions, and these connections occur primarily via the thalamus. In patients with bipolar disorder, regardless of age, neuroimaging studies have consistently reported an increased number of white matter hyperintensities, indicating possible alterations in striatum thalamus and thalamus-prefrontal cortex connections. METHODS: In the current study, we acquired high-resolution magnetic resonance imaging (MRI) and diffusion tensor (DT) scans of 40 patients with bipolar spectrum (BPS) illnesses (bipolar type I = 17, bipolar type II = 7, cyclothymia = 16) and 36 sex- and age-matched control subjects. Two researchers, without knowledge of diagnosis, outlined the caudate, putamen, and thalamus on contiguous axial MRI slices. We measured the volumes of the basal ganglia, thalamus, and gray/white matter of the frontal cortex. RESULTS: Bipolar spectrum patients as a single group did not differ from control subjects in thalamus and the basal ganglia volumes, but the cyclothymia patients had reductions in the volumes of putamen and the thalamus compared with control subjects. The BPS patients had significantly reduced volume of the white and the gray matter of the frontal cortex. Furthermore, compared with control subjects, BPS patients as a group showed alterations in anisotropy of the internal capsule adjacent to the striatum and thalamus and the frontal white matter. CONCLUSIONS: Our findings indicate that BPS patients may have distinct anatomical alterations in brain structures involved in the regulation of mood and cognition, as well as alterations in these structures' connection to related brain areas. PMID- 15820231 TI - Amphetamine sensitization impairs cognition and reduces dopamine turnover in primate prefrontal cortex. AB - BACKGROUND: Amphetamine (AMPH) sensitization in monkeys produces long-lasting behavioral changes that model positive (hallucinatory-like behaviors) and negative (psychomotor depression) symptoms of schizophrenia. The extent to which this model produces the core deficit in schizophrenia--working memory impairment- is unknown. METHODS: Two groups of rhesus monkeys were sensitized to AMPH over 6 weeks. In one group, acquisition of cognitive tasks (delayed response, visual discrimination, delayed nonmatch-to-sample) was examined beginning 6+ months postsensitization. The second group was pretrained to stability on delayed response before sensitization. Regional postmortem concentrations of dopamine and its metabolites were examined in tissue from age-matched AMPH-naive and AMPH sensitized monkeys using high-performance liquid chromatography with electrochemical detection (HPLC-ECD). RESULTS: The AMPH-sensitized monkeys were profoundly impaired in their ability to acquire cognitive tasks compared with AMPH-naive monkeys. Pretrained monkeys showed impaired delayed response performance for several months following sensitization. Analysis by HPLC revealed that AMPH sensitization significantly reduced dopamine turnover in prefrontal cortex and striatum. CONCLUSIONS: Impairments in the acquisition and performance of spatial delayed response in association with reduced dopamine turnover in prefrontal cortex following AMPH sensitization provide further support for the relevance of this model to both the etiology and the treatment of cognitive dysfunction in schizophrenia. PMID- 15820232 TI - Cortical and subcortical brain effects of transcranial magnetic stimulation (TMS) induced movement: an interleaved TMS/functional magnetic resonance imaging study. AB - BACKGROUND: To date, interleaved transcranial magnetic stimulation and functional magnetic resonance imaging (TMS/fMRI) studies of motor activation have not recorded whole brain patterns. We hypothesized that TMS would activate known motor circuitry with some additional regions plus some areas dropping out. METHODS: We used interleaved TMS/fMRI (11 subjects, three scans each) to elucidate whole brain activation patterns from 1-Hz TMS over left primary motor cortex. RESULTS: Both TMS (110% motor threshold) and volitional movement of the same muscles excited by TMS caused blood oxygen level-dependent (BOLD) patterns encompassing known motor circuitry. Additional activation was observed bilaterally in superior temporal auditory areas. Decreases in BOLD signal with unexpected post-task "rebounds" were observed for both tasks in the right motor area, right superior parietal lobe, and in occipital regions. Paired t test of parametric contrast maps failed to detect significant differences between TMS- and volition-induced effects. Differences were detectable, however, in primary data time-intensity profiles. CONCLUSIONS: Using this interleaved TMS/fMRI technique, TMS over primary motor cortex produces a whole brain pattern of BOLD activation similar to known motor circuitry, without detectable differences from mimicked volitional movement. Some differences may exist between time courses of BOLD intensity during TMS circuit activation and volitional circuit activation. PMID- 15820233 TI - Event-related functional magnetic resonance imaging investigation of executive control in very old individuals with mild cognitive impairment. AB - BACKGROUND: Attentional control of executive cognitive function (ECF) decreases in older individuals with Alzheimer Disease (AD). In order to examine early AD related changes in the neural substrates of ECF attentional control, we measured activation dorsolateral prefrontal (dLPFC), posterior parietal (PPC), and anterior cingulate cortex (ACC) in adults with mild cognitively impairment (MCI) and in cognitively normal (CN) adults. METHODS: Functional magnetic resonance imaging analysis of brain activation in MCI (n = 8, mean age 79.5) and CN (n = 8 mean age 81.5) during increasing loads of attentional demands. RESULTS: MCI and CN older adults performed with similar accuracy and reaction time. MCI had greater activation than CN in PPC (right p = .03 and left p = .05) and dlPFC areas (right p = .002 and left p = .004), while activation in ACC was similar in the two groups. Response to increasing loads of the task differed by group: MCI selectively engaged bilateral PPC (right p = .03, left p = .04), while CN subjects increased bilateral dlPFC activation (right p = .005 and left p = .02) and ACC activation (p = .04). Among MCI, greater load-related changes in PPC activity were associated with smaller load-related changes in accuracy rates (r = -.85, p = .07) and greater increases in reaction times (r = .97, p = .01). In CN subjects, load-related change in PPC activation was associated with load-related change in reaction time (r = .76, p = .02) but not with changes in accuracy rates. CONCLUSIONS: PPC and dlPFC may show early functional changes associated with MCI. PMID- 15820234 TI - Striatal and forebrain nuclei volumes: contribution to motor function and working memory deficits in alcoholism. AB - BACKGROUND: Striatal structures are involved in dopaminergic alcohol reward mechanisms and aspects of motor control. Basal forebrain structures hold cholinergic mechanisms influencing memory formation, vulnerable to chronic alcoholism; however, alcoholism's effect on volumes of these structures has seldom been considered with in vivo measurement. METHODS: We measured bilateral volumes of caudate nucleus, putamen, nucleus accumbens, and medial septal/diagonal band (MS/DB) in 25 men with alcohol dependence and 51 age-matched control men. Six alcoholic subjects had been drinking recently, and 19 had been sober. RESULTS: Volumes of caudate and putamen were smaller in the alcoholics than in the control subjects, regardless of length of sobriety. Recent drinkers showed greater deficits in nucleus accumbens than sober alcoholics. Putamen volume was positively correlated with grip strength; MS/DB volume was positively correlated with verbal working memory independently of the negative association between age-standardized MS/DB and age in alcoholics. CONCLUSIONS: Caudate and putamen volume deficits occur and endure in chronic alcoholism. Nucleus accumbens might be especially sensitive to recent alcohol exposure. Striatal volumes should be considered in functional imaging studies of alcohol craving that target striatal brain regions. The age-alcohol interaction for MS/DB volumes is consistent with a cholinergic mechanism for the working memory impairment observed in the alcoholics. PMID- 15820235 TI - Reelin signaling is impaired in autism. AB - BACKGROUND: Autism is a severe neurodevelopmental disorder with genetic and environmental etiologies. Recent genetic linkage studies implicate Reelin glycoprotein in causation of autism. To further investigate these studies, brain levels of Reelin protein and mRNA and mRNAs for VLDLR, Dab-1, and GSK3 were investigated. METHODS: Postmortem superior frontal, parietal, and cerebellar cortices of age, gender, and postmortem interval-matched autistic and control subjects were subjected to SDS-PAGE and Western blotting of Reelin protein. Quantitative reverse transcriptase polymerase chain reaction analysis of Reelin, VLDL-R, Dab-1, and GSK3 mRNA species in superior frontal and cerebellar cortices of autistic and control subjects were also performed. RESULTS: Reelin 410, 330, and 180 kDa/beta-actin values were reduced significantly in frontal and cerebellar, and nonsignificantly in parietal, areas of autistic brains versus control subjects, respectively. The mRNAs for Reln and Dab-1 were reduced significantly whereas the mRNA for Reln receptor VLDLR was elevated significantly in superior frontal and cerebellar areas of autistic brains versus control brains, respectively. CONCLUSIONS: Reductions in Reelin protein and mRNA and Dab 1 mRNA and elevations in Reln receptor VLDLR mRNA demonstrate impairments in the Reelin signaling system in autism, accounting for some of the brain structural and cognitive deficits observed in the disorder. PMID- 15820236 TI - Antibiotic prophylaxis with azithromycin or penicillin for childhood-onset neuropsychiatric disorders. AB - BACKGROUND: The acronym PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections) describes a subgroup of children with obsessive-compulsive disorder and/or tic disorder that experience symptom exacerbations following streptococcal infections. We hypothesized that the prevention of streptococcal infections among children in the PANDAS subgroup would decrease neuropsychiatric symptom exacerbations. METHODS: Twenty-three subjects with PANDAS were enrolled in a double blind, randomized controlled trial. Antibiotic prophylaxis with penicillin or azithromycin was administered for 12 months. Rates of streptococcal infections and neuropsychiatric symptom exacerbations were compared between the study year and the baseline year prior to entry. RESULTS: Significant decreases in streptococcal infections during the study year were found with a mean of .1 (.3 SD) per subject, compared to the baseline year with 1.9 (1.2 SD) in the penicillin group and 2.4 (1.1 SD) in the azithromycin group [p<.01]. Significant decreases in neuropsychiatric exacerbations during the study year were also found with a mean of .5 (.5 SD) per subject in the penicillin group and .8 (.6 SD) in the azithromycin group, compared to the baseline year with 2.0 (.9 SD) in the penicillin group and 1.8 (.6 SD) in the azithromycin group [p<.01]. CONCLUSIONS: Penicillin and azithromycin prophylaxis were found to be effective in decreasing streptococcal infections and neuropsychiatric symptom exacerbations among children in the PANDAS subgroup. PMID- 15820237 TI - Bupropion XL in adults with attention-deficit/hyperactivity disorder: a randomized, placebo-controlled study. AB - BACKGROUND: Data remain limited on treatment strategies for adults with attention deficit/hyperactivity disorder (ADHD). This study evaluated the efficacy and safety of an extended-release, once-daily formulation of bupropion (XL) in the treatment of adults with ADHD. METHODS: This multisite, placebo-controlled, 8 week prospective trial evaluated 162 adult patients diagnosed with ADHD (combined and inattentive types). Subjects were treated with up to 450 mg/day of bupropion XL. The primary efficacy endpoint was the proportion of ADHD responders (defined as at least a 30% reduction in the investigator-rated ADHD Rating Scale score) at week 8 (last observation carried forward [LOCF]). RESULTS: Bupropion XL responders (53%) exceeded placebo responders (31%) (p =.004 at week 8) with a significantly greater proportion of bupropion XL responders as early as week 2 (p = .01). Treatment effect size calculated for the ADHD Rating Scale total score was .6. Bupropion XL appeared to provide sustained benefit throughout the day compared with placebo (morning p =.033, afternoon p =.004, evening p = .024). Bupropion XL was safe and well tolerated, with no serious or unexpected adverse events and a low rate of drug-related study discontinuation (5%). CONCLUSIONS: The results from this multisite study indicate that bupropion XL is an effective and well-tolerated nonstimulant treatment for adult ADHD. PMID- 15820238 TI - Smoking cue reactivity in schizophrenia: effects of a nicotinic receptor antagonist. AB - BACKGROUND: Rates of cigarette smoking in schizophrenia are higher than in the general population. To investigate differences in sensitivity to smoking cues between schizophrenia and control subjects, we compared smoking cue reactivity (CR) in schizophrenia versus control smokers with and without pretreatment with the nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine (MEC). METHODS: Smoking CR in schizophrenia (n = 22) and nonpsychiatric control (n = 20) smokers was determined using exposure to smoking pictures. Three doses of MEC (0, 5, and 10 mg/day) were administered during the 3 test weeks to determine the role of nAChRs in mediating the smoking CR response. RESULTS: Eleven of 22 (50%) schizophrenia and 10 of 20 (50%) control smokers displayed smoking CR. Smoking CR was not significantly different between schizophrenia and control smokers in the placebo (0 mg/day) condition. However, MEC pretreatment produced a dose-dependent reduction of CR in schizophrenia smokers compared with placebo. There was no significant effect of MEC on CR in control smokers. CONCLUSIONS: Our findings suggest that blockade of CR by MEC may be more robust in schizophrenia versus control smokers, possibly due to reduced nAChR levels in the brains of patients with schizophrenia. PMID- 15820239 TI - Activation in dorsolateral prefrontal cortex in response to maternal criticism and praise in recovered depressed and healthy control participants. AB - BACKGROUND: High family levels of expressed emotion reliably predict relapse in patients with schizophrenia and mood disorders; however, the neural mechanisms linking expressed emotion and relapse are unexplored. Dysfunctional activity in the dorsolateral prefrontal cortex (DLPFC) has been implicated in the pathophysiology of depression. Functional magnetic resonance imaging (fMRI) was used to assess focal activation changes in DLPFC in response to a novel psychosocial challenge stimulus developed from the expressed emotion construct. METHODS: Healthy control subjects and fully remitted unipolar depressed participants completed blood oxygen level-dependent fMRI while they heard their own mothers making critical and praising comments about them. RESULTS: Relative to control subjects, participants with a history of depression failed to activate DLPFC when they heard critical remarks. There were no differences between the two groups in their DLPFC responses to maternal praise. CONCLUSIONS: Even if fully well at the time of testing, participants with a known vulnerability to depression respond differently to the psychosocial challenge of being criticized. These findings might have implications for our understanding of vulnerability to depression and to depressive relapse. PMID- 15820240 TI - Decreased serum amyloid beta(1-42) autoantibody levels in Alzheimer's disease, determined by a newly developed immuno-precipitation assay with radiolabeled amyloid beta(1-42) peptide. AB - BACKGROUND: Autoantibodies against amyloid beta (A beta) peptide found in patients with Alzheimer's disease (AD) also occur naturally in the general population independently of the cognitive status. METHODS: We compared serum A beta(1-42) autoantibody levels (A beta(1-42)-AL) of 96 AD patients and 30 healthy elderly control subjects (HC), assessing their diagnostic value for AD with a newly developed immunoprecipitation assay with radiolabeled A beta(1-42) peptide. RESULTS: We found a highly significant decrease of A beta(1-42)-AL in AD patients (p = .001) independently of age, cognitive status, and apolipoprotein E epsilon4 carrier status. Amyloid beta(1-42) autoantibody levels were correlated with gender in AD, with a higher level occurring in women. When A beta(1-42) autoantibody sensitivity (specificity) was set >80%, specificity (sensitivity) was below 50% to correctly allocate patients and healthy control subjects. CONCLUSIONS: Our data indicate a potentially pathophysiologic decrease of serum A beta(1-42) antibodies in AD. Amyloid beta(1-42) antibodies in the serum alone, however, seem not to be useful as a diagnostic marker of AD. PMID- 15820241 TI - Female schizophrenia patients have prepulse inhibition deficits. AB - BACKGROUND: Prepulse inhibition (PPI) of startle shows sexual dimorphism: women have lower levels of PPI than do men, and have menstrual cycle shifts in PPI. Many studies report PPI deficits in male schizophrenia patients; one recent report identified PPI deficits in male but not female patients. This study was designed to determine whether female schizophrenia patients have lower levels of PPI than normal females. METHODS: Twenty-five female schizophrenia patients, and 26 normal females were tested in a startle paradigm using 115 dB startle pulses and prepulses of 8 and 16 dB above a 70 dB background, with 30 and 120 msec prepulse intervals. RESULTS: Female patients had significantly less PPI compared with normal females, particularly when 16 dB prepulses were utilized. Patients also exhibited a nonsignificant trend towards lower levels of habituation compared to normal subjects. CONCLUSIONS: Under the present paradigmatic and subject acquisition conditions, female schizophrenia patients had PPI deficits compared to normal females. PMID- 15820242 TI - Temporal changes in the management of diverticulitis. PMID- 15820243 TI - The reduced tolerance of rat fatty liver to ischemia reperfusion is associated with mitochondrial oxidative injury. AB - BACKGROUND: Oxidative stress contributes to the pathogenesis of hepatic ischemia reperfusion injury. This study aimed to determine whether fatty degeneration affects the oxidative damage during warm ischemia reperfusion and whether mitochondria, the major intracellular site of energy synthesis, represent a preferential target of this injury. MATERIALS AND METHODS: Fed rats with control or fatty liver induced by choline deficiency underwent 60' lobar ischemia and reperfusion. Oxidative damage was assessed by measuring in whole liver tissue and in isolated mitochondria the thiobarbituric acid-reactive substances (TBARs), protein carbonyls (PC), and total and oxidized glutathione (GSH and GSSG) concentrations. The mitochondrial F0-F1-ATPase content and the oxidative phosphorylation activity were also determined. Rat survival and ALT release were assessed as parameters of liver injury. RESULTS: In the whole liver tissue, with the exception of TBARs, no differences were observed for GSH, GSSG, and PC between the two groups throughout all of the experiment. In contrast, in isolated mitochondria, fatty infiltration was associated with a mild oxidative imbalance already under basal conditions. The preischemic differences in the mitochondrial TBARs, PC, and GSSG levels were significantly amplified by reperfusion in the presence of steatosis. The enhanced oxidative damage was associated to a reduced F0-F1-ATPase content and oxidative phosphorylation activity in fatty liver mitochondria. Finally, serum ALT levels were significantly greater and survival significantly lower in rats with steatotic liver. CONCLUSIONS: Fatty infiltration exacerbates mitochondrial oxidative injury during warm ischemia reperfusion. The increased oxidative stress can alter mitochondrial functions, including key processes for ATP synthesis, thus, contributing to the reduced tolerance to reperfusion injury. PMID- 15820244 TI - Is it reasonable to add preoperative serum level of CEA and CA19-9 to staging for colorectal cancer? AB - BACKGROUND: Carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) are the most common tumor markers for colorectal cancer. The aim of this study was to evaluate the possibility of adding them into the current staging system by analyzing their prognostic significance. MATERIALS AND METHODS: The study population was patients (n = 574, 67.1 +/- 11.3 years old, 397 males) who received potentially curative resection of colorectal adenocarcinoma (stage I III) between January 1994 and August 2002, including preoperative measurements of CEA and CA19-9. Clinicopathological characteristics and associated follow-up data were retrospectively collected by reviewing available medical charts. CEA higher or equal to 5 ng/ml was defined as abnormal (CEA+). The CA19-9 level was set at 37 U/ml (CA19-9+). Patients were further divided into four groups (1, 2, 3, 4) according to the results of these two markers (CEA/CA19-9: -/-, -/+, +/-, and +/+). Survival was analyzed for AJCC staging, CEA (+) versus (-), CA19-9 (+) versus (-), and four groups. RESULTS: CEA and CA19-9 survival curves were not significantly different. However, the combined use of the two markers revealed a significant survival benefit (P = 0.035) of group 1 ("-" for both markers) over 4 ("+" for both) in stage II. CONCLUSIONS: Patients with an elevated level of both CEA and CA19-9 in stage II of colorectal cancer have a significantly poorer prognosis than those with normal levels of these markers. We recommend adding both CEA and CA19-9 to the current staging system. PMID- 15820245 TI - The influence of venous drainage on autologous ovarian transplantation. AB - BACKGROUND: Despite many investigations of ovarian transplantation, it is still difficult to define the best site for ovarian implantation, considering its endocrine function. We sought to evaluate the influence of venous drainage on ovarian physiomorphological aspects and to assess ovary transplantation by intact and sliced techniques. MATERIALS AND METHODS: One hundred and eight cycling female Wistar rats were randomly divided into six groups (n = 18): G1 (control, sham operation), G2 (bilateral oophorectomy), G3 (transplantation of intact ovarium on the greater omentum), G4 (transplantation of sliced ovarium on the greater omentum), G5 (transplantation of intact ovarium in the retroperitoneum), G6 (transplantation of sliced ovarium in the retroperitoneum). The blood drainage of groups G3 and G4 was to the portal vein and that of groups G5 and G6 was to the cava. The estrus cycle was investigated in the 3rd and 6th months. Serum estradiol concentration was also assessed in all groups in the 3rd and 6th months. Histological studies of the transplanted ovaria were carried out at that time. RESULTS: G1 animals preserved the cycling sequence. G2 rats persisted in diestrus. Nine G3 rats showed estrus cycles. Fourteen G4 rats had normal cycles and 1 rat persisted in estrus. Seven G5 rats showed estrus vaginal smears. Nine G6 animals had estrus vaginal smear patterns and 3 showed incomplete cycles in the 3rd month and complete in the 6th month. The histology of all ovaria confirmed the preservation of their vitality. The ovaries of G5 and G6 animals had more cysts and fewer corpora lutea. The sliced ovaries implanted on the omentum presented higher estradiol concentrations. CONCLUSIONS: The ovaria that drained to the portal vein presented a better physiomorphological aspect than those that drained to the cava. The sliced ovaria were more efficient than the intact ones. PMID- 15820246 TI - Hypoxia augments gelatinase activity in a variety of adenocarcinomas in vitro. AB - BACKGROUND: Hypoxia within solid adenocarcinomas and protease up-regulation has been independently implicated as poor prognostic indicators in a variety of tumor types. The authors hypothesize that Matrix Metalloproteases (MMP) are up regulated in direct response to a hypoxic environment. MATERIALS AND METHODS: Colonic (SW1222), breast (MDA-MB231), and pancreatic (PSN-1) tumor cell lines were exposed to hypoxia (1% oxygen/94% nitrogen/5% carbon dioxide) for periods of up to 24 h. Reaction to a hypoxic environment was determined via invasion across a Matrigel-coated 8-microm Transwell filter. Activity of MMP 2 and 9 was assessed using gelatin zymography. Expression of tissue inhibitor of metalloproteases 1 (TIMP-1) was quantified using ELISA (Biotrak). Correlation between protease expression and invasive capacity was determined using a specific gelatinase inhibitor (MMPI; Calbiochem). RESULTS: All tumor lines demonstrated augmented invasion over 72 h (P < 0.01 all groups). Concomitant significant increase in MMP 2 and 9 activity was observed in the SW1222 and PSN-1 lines. MDA-MB231s showed increase in MMP 9 expression and in a unidentified 103-kDa gelatinase (P < 0.001). The hypoxia-augmented invasion was attenuated by the addition of a specific gelatinase inhibitor confirming interdependence. CONCLUSIONS: Hypoxia induces an increased invasive capacity via gelatinase up-regulation without loss of cell viability. This suggests a mechanism explaining the poorer prognosis seen in patients with protease-secreting solid adenocarcinomas. PMID- 15820247 TI - Effects of total parenteral nutrition on rat enteric nervous system, intestinal morphology, and motility. AB - Total parenteral nutrition (TPN) is often crucial for patients not being able to feed enterally or having intestinal absorptive deficits. Enteral nutrition is, however, frequently regarded vital for maintaining functional and structural intestinal integrity. The aim of this study was to investigate possible effects of TPN on rat distal small intestine compared to enterally fed identically housed controls, regarding the enteric nervous system (ENS), motility in vitro, and morphology. This study shows that motor responses evoked by electrical stimulation or exposure to vasoactive intestinal peptide (VIP), pituitary adenylate cyclase activating peptide-27 (PACAP-27), and nitric oxide (NO) donor were unchanged. By using immunohistochemistry, the numbers of submucous (P < 0.05) and myenteric (P < 0.05) nerve cells were found to increase, expressed as numbers per unit length. The percentage of neurons expressing VIP, PACAP-27, NO synthase, and galanin remained unchanged, however. By in situ hybridization the number of submucous neurons expressing neuropeptide Y-mRNA was found to decrease (P < 0.05); the other populations were unaltered. Morphometry revealed an increased submucosal thickness (P < 0.05), while intestinal circumference markedly decreased (P < 0.0001) in TPN-treated rats. In conclusion, TPN treatment resulted in reduced intestinal circumference leading to condensation of enteric neurons. No marked changes in neurotransmitter expression of the enteric neurons or in motor activity were noted. PMID- 15820248 TI - Caspase-3 inhibition preserves myocardial geometry and long-term function after infarction. AB - BACKGROUND: Cardiac remodeling after infarction is characterized by progressive ventricular dilation and functional impairment. Although surgical and percutaneous revascularization strategies may prevent remodeling, not all patients are candidates for these procedures. Apoptosis in the border-zone myocardium is thought to be critical to the remodeling process, and caspase-3 is a downstream effector of apoptosis. We hypothesized that inhibition of caspase-3 activity might limit dysfunction and remodeling after permanent coronary artery ligation. METHODS: FVB male mice underwent permanent ligation of the left anterior descending coronary artery. Immediately before surgery and for 7 subsequent days, animals were treated daily with 3 mg/kg DEVD-CHO, a cell permeable inhibitor of caspase-3 (n = 16), or vehicle (n = 28). At 2 weeks and 6 weeks, echocardiography was performed and ventricular dimensions and function were assessed. At 8 weeks, invasive hemodynamic measurements were made and the animals were sacrificed. RESULTS: There was a trend toward improved survival in the inhibitor-treated group compared to the vehicle-treated group (56.3% versus 21.4%, P = 0.07). Infarct size at the time of sacrifice was comparable in both groups. At both 2 and 6 weeks, left ventricular dimensions, including end diastolic and end-systolic diameters, were less in inhibitor-treated animals. Fractional shortening was higher at 6 weeks in the inhibitor-treated animals (22.1 +/- 6.0% versus 15.0 +/- 6.0%, P = 0.02). Invasive hemodynamic parameters at 8 weeks were comparable, with the exception of diastolic blood pressure, which was less in the inhibitor group. CONCLUSION: Treatment with a caspase-3 inhibitor improved survival and prevented ventricular dilation and dysfunction after permanent coronary artery occlusion. PMID- 15820249 TI - Adult stem cell lines in regenerative medicine and reconstructive surgery. AB - In recent years, there has been a tremendous increase in the understanding of stem cell biology. The potential clinical applications lead to an extended interest in the use of stem cells in many medical disciplines. Multipotent adult stem cells seem to be almost comparable to embryonic stem cells with respect to their ability to differentiate into various tissues in vitro and in vivo, a function that has been termed "stem cell plasticity". In vivo experiments in rodents have shown that adult stem cells participate in tissue- and organ regeneration in almost all lesions. Although stem cell populations isolated from the bone marrow are usually a heterogeneous mix of different subpopulations, cloned adult stem cell lines from any source also show a broad spectrum of differentiation potential, e.g., osteogenesis, myogenesis, neurogenesis, or angiogenesis in wound healing. Angiogenesis in particular is a subject in tissue regeneration with tremendous implication in reconstructive surgery. This comprehensive plasticity makes it possible to use stem cell lines for biomedical research, tissue engineering, regenerative surgery, and organ repair. Adult stem cell lines are molecularly well defined with respect to transcription factors, active signal transduction pathways, and expression of receptors/ligand pairs. We performed experiments with adult stem cell lines, which are not subject to stem cell heterogeneity. Results obtained with stem cell lines can reliably be ascribed to the stem cell population under scrutiny. Adult stem cell lines can be obtained with the necessary quality and quantity also to study many effects of human stem cells in vitro and in vivo. In this paper, we summarize some of the tremendous therapeutic implications of adult stem cell lines in surgery and surgical research. PMID- 15820250 TI - Determination of operation time in colorectal diseases: preoperative chemotherapy application. AB - BACKGROUND: Our aim was to determine the time it takes for wound healing to return to normal in cases where patients have undergone preoperative chemotherapy. MATERIALS AND METHODS: Eighty-four Wistar-albino rats were included in the study. Twelve of them were placed in the control group (Group I), with no further drug administration. Another 12 rats were placed in a sham group (Group II) and were peritoneally injected with 1 cc of isotonic saline solution 5 days a month, for a period of 6 months. The remaining 60 rats were placed in five chemotherapy groups (Groups III-VII) and were administered 20 mg/kg 5 fluorouracil through peritoneal injection, 5 days a month for a period of 6 months. At the end of the sixth cure, 12 rats from the control (Group I), sham (Group II), and chemotherapy groups (Group III) were operated on, and an intestinal transsection was applied to the rectosigmoid junction, followed by one by-one anastomosis using 5/0 vicryl. Other groups (Groups IV-VI) with chemotherapy treatment were operated on at 1-week intervals and subjected to the same procedure. The subjects were reoperated on on the eleventh day. A full-layer 4 x 4 cm piece was removed from the abdominal wall containing the previous incision line at the middle, for tensile strength pressure measurements. In addition, a 4 cm colon segment was removed for bursting pressure measurements. Plasma albumin and tissue hydroxyproline levels were measured, and fibroblast numbers were counted in the sections prepared from the abdominal wall. RESULTS: The control and sham groups were found to be similar to each other with respect to all parameters measured (P > 0.05). Significant reductions were observed in all parameters in the early chemotherapy groups compared with the control and sham groups (P <0.05). All parameters measured in Groups V, VI, and VII were found to be similar to those in the control and sham groups (P <0.05). CONCLUSION: Wound healing is impaired in rats with chemotherapy, but following the second week after the chemotherapy, disrupted parameters return to their normal levels. PMID- 15820251 TI - Right- and left-sided colorectal cancers display distinct expression profiles and the anatomical stratification allows a high accuracy prediction of lymph node metastasis. AB - Accurate preoperative prediction of lymph node metastasis and degree of tumor invasion would facilitate an appropriate decision of the extent of surgical resection of cancers to reduce unnecessary complication or to minimize the risk of recurrence in patients. We analyzed gene expression profiles characteristic of the invasiveness of colorectal carcinoma in a total of 89 cases, using a cDNA array and pattern classification algorithms. We set binary classes for a panel of clinicopathologic parameters, each of which was divided at different levels for categories (discrete) or values (continuous). We searched an optimal combination of genes to discriminate the classes by using of a feature subset selection algorithm, which was applied to a set of genes preselected on the basis of statistical difference in expression (two-sided t test, P < or = 0.05). We used a sequential forward feature selection which additively searched a combination of genes, giving a minimal leave-one-out classification error rate of a k-nearest neighbor classifier. In the process of gene preselection, we found a remarkable difference in the expression pattern of genes according to the anatomical location of cancers. The difference was most prominent when the classes were set for cecum, ascending colon, transverse colon, and descending colon (CATD) versus sigmoid colon and rectum (SR). By stratifying these two locations, we were able to extract gene expression profiles characteristic of the classes of the presence versus absence of lymph node metastasis, lymphatic invasion, vascular invasion and degree of mural invasion, and pathological stages, with an accuracy of more than 90%. These results suggest that colorectal cancers harbor distinct molecular pathophysiological statuses according to their right-to-left locations, of which stratification is important for pattern classification of cDNA array data. PMID- 15820252 TI - Prediction of lymphatic invasion/lymph node metastasis, recurrence, and survival in patients with gastric cancer by cDNA array-based expression profiling. AB - BACKGROUND: We assessed the predictability of various classes of gastric carcinoma defined by clinicopathological parameters, such as invasiveness and clinical outcomes, using cDNA array data obtained from 54 cases. MATERIALS AND METHODS: We searched an optimal combination of genes to discriminate the classes defined with the clinicopathological parameters by using a feature subset selection algorithm, which was applied to a set of genes preselected on the basis of statistical difference in expression (two-sided t test, P < or = 0.05). With the selected features (gene set), we evaluated the predictability of each parameter in a leave-one-out cross-validation test. RESULTS: We successfully selected sets of genes for which the classifier predicted better versus worse overall survival (tumor-specific death) and tumor-free survival (recurrence), with respective classification rates of 94 and 92%. A contingency table analysis (chi2 test) and Cox proportional hazard model analysis revealed that lymph node metastasis is the most important factor (confounding factor) in patients' prognoses and risks of recurrence. The feature subset selection procedure successfully extracted expression patterns characteristic of lymph node metastasis and lymphatic vessel invasion, yielding 92 and 98% prediction accuracies for these respective factors. CONCLUSION: We conclude that expression profiling using feature subset selection provides a powerful means of stratification of gastric cancer patients in regard to the prognostic factors. Further studies should be warranted to apply this method to personalization of the treatment options. PMID- 15820253 TI - Effect of jaundice and its resolution on wound re-epithelization, skin collagen synthesis, and serum collagen propeptide levels in patients with neoplastic pancreaticobiliary obstruction. AB - BACKGROUND: Wound and anastomotic healing is considered to be delayed in patients with obstructive jaundice. The study was designed to find out whether the healing of experimental suction blister wounds, skin collagen synthesis, and serum procollagen levels are affected by obstructive jaundice, and if biliary drainage may cause any alterations in these processes. PATIENTS AND METHODS: Suction blisters were induced on 24 patients with obstructive jaundice caused by neoplastic pancreaticobiliary obstruction and 17 control patients with the corresponding condition without jaundice, to compare healing parameters and collagen synthesis between the groups. A second set of suction blisters were induced on 13 formerly jaundiced patients after the resolution of jaundice and on 14 control patients, to find out whether drainage or time modifies healing or collagen synthesis. By using this model, it is possible to evaluate the re epithelization and inflammation on wound healing and to assess the baseline skin collagen synthesis. The healing of suction blisters was followed up by measuring water evaporation and blood flow in the wound. Blister fluids and serum samples were collected to study collagen propeptides. RESULTS: Healing of the blister wound was unaffected by obstructive jaundice. Drainage had no effect on healing. The baseline synthesis of type I and type III collagen in the skin was decreased in jaundiced patients. Biliary drainage improved the synthesis. Serum type III procollagen propeptide levels were elevated in jaundiced patients, but began to normalize after drainage. CONCLUSION: Healing of an experimental blister wound is not disturbed by obstructive jaundice. The decreased baseline skin collagen synthesis is partly restored by the resolution of jaundice. The results indicate that cell protein synthesis is disturbed earlier than cell dynamics in obstructive jaundice. The elevated serum PIIINP levels, which are most likely to be related to early fibrosis in liver, decreased after drainage. PMID- 15820254 TI - Hepatic expression of S32A/S36A IkappaBalpha does not reduce postischemic liver injury. AB - BACKGROUND: Activation of the transcription factor, NF-kappaB, during hepatic ischemia/reperfusion injury is associated with proinflammatory mediator expression and is thought to be one of the initial triggers for the inflammatory response after reperfusion. In the current study, we sought to determine whether in vivo adenoviral transfection of a mutant inhibitor of kappaB-alpha (IkappaBalpha), which cannot be serine phosphorylated or degraded (IkappaBalphaSR), would inhibit NF-kappaB and ameliorate the hepatic inflammatory response to ischemia/reperfusion. MATERIALS AND METHODS: Male C57BL/6 mice were subjected to sham surgery or partial hepatic ischemia (90 min) and reperfusion (up to 8 h). Mice were infected with 1 x 10(9) PFU of adenovirus containing either beta-galactosidase (LacZ) or IkappaBalphaSR 3 days prior to induction of ischemia. Serum and tissues were obtained at various times for analysis. RESULTS: In unmanipulated mice, degradation of IkappaBalpha, as occurs after serine phosphorylation, was evident in liver by the end of ischemia and during early reperfusion. Mice transfected with IkappaBalphaSR displayed the same degree of inflammation and hepatocellular injury as LacZ-transfected mice. There was no difference between LacZ- and IkappaBalphaSR-transfected livers in terms of NF kappaB activation or proinflammatory cytokine production. CONCLUSIONS: The data demonstrate that the pathway of NF-kappaB activation involving serine phosphorylation of IkappaBalpha is not the primary mechanism for induction of liver inflammation after ischemia/reperfusion and suggest that alternative pathways, such as tyrosine phosphorylation of IkappaBalpha, may be essential for the postischemic response in liver. PMID- 15820255 TI - Uretero-neocystostomy in a swine model of kidney transplantation: a new technique. AB - BACKGROUND: Swine constitute a well-characterized large animal model for kidney transplantation (K-Tx) although the uretero-neocystostomy represents a crucial aspect because of the very small caliber of the ureter and the mucosal susceptibility to the edema during surgical management. Besides infectious and occlusive complications limited the employment of a stent and the peculiar anatomy of the species prevents its removal without an operative approach. These features find an equivalent in children (<4 year old) candidates to urological surgery. We investigated an uretero-neocystostomy technique to be applied in case of narrow caliber without using endoluminal device. MATERIALS AND METHODS: Thirty unrelated large-white piglets, divided into three equal groups (n = 10) on the basis of the uretero-neocystostomy technique used, underwent K-Tx while receiving a double immunosuppressive regimen. We developed a direct uretero-neocystostomy (Direct) technique and compare this with the gold standard Lich-Gregoir (LG) and Leadbetter-Politano (LP) techniques. RESULTS: After 60 days, the ureteral complication rates were: group 1 (LG): 60% (6/10, three early and three late strictures); group 2 (LP): 40% (4/10, two leakages, one early stricture, and one isolated ureteral dilatation); group 3 (Direct): 10% (1/10, one isolated ureteral dilatation). No bacterial cystitis or nephritis occurred; retrograde cystograms were negative for refluxes in every group. CONCLUSIONS: The comparison between the old techniques and the new one (LG and LP versus Direct) showed a lower incidence of complications among the animals that underwent the new direct technique (P < 0.05). This technique could be applied clinically in selected pediatric cases either of transplantation or reflux. PMID- 15820256 TI - Inhibition of geranylgeranyltransferase I decreases generation of vascular reactive oxygen species and increases vascular nitric oxide production. AB - BACKGROUND: Vascular injury with endothelial dysfunction results in an imbalance between the production of vasoprotective molecules such as nitric oxide (NO) and deleterious reactive oxygen species (ROS). The purpose of this work was to test the hypothesis that inhibition of geranylgeranyltransferase I (GG Tase I) reduces vascular injury by increasing vascular NO production while decreasing ROS generation. METHODS AND RESULTS: GGTI-298 decreased the formation of intimal hyperplasia at 14 days following balloon injury. GGTI-298 (10 microm) inhibited activation of RhoA and Rac1 as well as inhibited SMC proliferation. GGTI increased SMC-inducible NO synthase (iNOS) levels and NO production in vitro. Additionally, the activation of NAD(P)H oxidase subunits was decreased by GGTI in vitro. This correlated with a decrease in TNF-alpha- or angiotensin-II-induced ROS production assayed by DCF fluorescence. In vivo, GGTI treatment increased endothelial NOS (eNOS) expression in uninjured arteries and iNOS expression in balloon-injured arteries. Furthermore, GGTI treatment attenuated balloon-injury induced superoxide generation assayed by MCLA luminescence. CONCLUSIONS: GGTI decreases the production of ROS and increases the production of NO both in vitro and in vivo. These effects may be mediated via the inhibition of activation of the small GTPases Rac1 and RhoA. Pharmacological inhibition of GGTase I may prove to be a useful clinical adjunct in the treatment of cardiovascular diseases. PMID- 15820257 TI - Cotransfection of DC with TLR4 and MART-1 RNA induces MART-1-specific responses. AB - BACKGROUND: Cotransfection of dendritic cells (DC) with MART-1 and constitutively active TLR4 (caTLR4) RNA enhances the maturation of DC. MATERIALS AND METHODS: Immature DC were cotransfected with RNA constructs encoding MART-1 and caTLR4, and CTL responses were analyzed. RESULTS: Cotransfection of DC with MART-1 + caTLR4 enhanced the expression of CD80 and CD83 surface markers and increased the secretion of cytokines IL-6, IL-12, and TNFalpha. Neither the native nor the A27L modified MART-1 RNA could induce significant DC maturation or cytokine secretion. More importantly, DC cotransfected with caTLR4 + MART-1 RNA induced MART-1 specific CTL responses of a higher magnitude than DC transfected with either the native or A27L MART-1 RNA. When the MART-1 RNA-transfected DC were treated with DC-maturing cytokines, the induced CTL were less frequent and less lytic than those induced with MART-1 + caTLR4. A 2- to 100-fold increase in MART-1 tetramer+ cells and 2- to 10-fold increases in IFNgamma secretion and cytotoxicity were seen in CTL induced with MART-1 + caTLR4 compared to CTL induced with either MART 1 or A27L RNA. CTL induced with the mixed RNA displayed high percentages of CD8+ cells coexpressing CD45RA, CD56, and 2B4 antigens. Transfection with caTLR4 alone induced DC maturation, but did not induce lytic CTL, suggesting that CTL responses were induced solely by MART-1 epitopes. CONCLUSIONS: caTLR4 increases the CTL-inducing capacity of DC generating a lytic response specific for the accompanying antigen. These results demonstrate the possibility of enhancing the immunogenicity of the native MART-1 and other RNA derived from weakly immunogenic tumors in DC-based immunotherapy. PMID- 15820258 TI - Photochemical repair of Achilles tendon rupture in a rat model. AB - BACKGROUND: Photochemical tissue bonding (PTB) is an emerging technique for bonding or sealing tissue surfaces that requires light and a photoactive dye for its effect. The potential of PTB for tendon repair was assessed in a rat model. MATERIALS AND METHODS: The optical properties of bovine tendon were determined ex vivo to gauge the depth of light penetration as a function of wavelength and dosimetry parameters were established for PTB repair of ruptured tendon. PTB was then tested in vivo to repair transected tendons in Sprague-Dawley rats. Repair strengths were measured using a strain gauge up to 14 days post treatment. RESULTS: The effective penetration depth in tendon was estimated to be 0.68 mm at 514 nm. Following PTB treatment of mechanically ruptured tendon, significant bonding was dependent on the presence of both light and dye and attained a plateau strength at a fluence of 125 J/cm2. In a subsequent in vivo study to investigate PTB for repair of transected rat Achilles tendon, the ultimate stress required to break the repaired tendon was measured immediately after irradiation and at 7 and 14 days post-repair. Results showed that the difference in the ultimate stress between control and PTB treatment groups was statistically significant immediately after treatment and at 7 days (p = 0.04) but not 14 days (p = 0.75) post-repair. CONCLUSIONS: PTB provides a benefit to tendon repair at early stages in repair and is worthy of further investigation as a potential surgical adjunct for tendon repair in orthopedic surgeries. PMID- 15820259 TI - A technique of bone marrow collection from vertebral bodies of cynomolgus macaques for transplant studies. AB - BACKGROUND: Strategies to induce donor-specific allograft tolerance are best tested in preclinical models developed in nonhuman primates (NHPs). Most protocols prepare the recipient by infusing hematopoietic cells from the donor. We report here a procedure to isolate and characterize large numbers of bone marrow cells (BMCs) from cynomolgus monkeys (cynos) that can then successfully be transplanted into conditioned recipients. MATERIALS AND METHODS: Vertebral columns of five cynos were excised en bloc and separated into individual vertebrae. The cancelous bone was extracted with a core puncher, fractionated, filtered, centrifuged, and resuspended in transplantation media before being analyzed by flow cytometry. In two instances, the collected BMCs were reinfused into allogeneic recipients preconditioned with a nonmyeloablative regimen. Chimerism was monitored using short-tandem repeat analysis. RESULTS: The mean total BMCs yield was 25.5 x 10(9) (range of 4.00 x 10(9) to 59 x 10(9)) with mean cell viability of 93.4% (range: 90-96%). CD34+ cells and CD3+ cells averaged 0.34 and 3.91% of total BMCs, respectively. This resulted in absolute cell number yields of 1.02 x 10(8) and 1.15 x 10(9) for CD34+ and CD3+ cells, respectively. Graft-versus-host disease was absent in both bone marrow infused animals, and a maximum level of chimerism of 18% was detected at 3 weeks after BMCs infusion. CONCLUSION: We present here the first detailed report of a procedure to retrieve and characterize large numbers of BMCs from vertebral bodies of cynos and demonstrate that cells collected with this technique have the capability of engrafting in allogenic recipients. PMID- 15820260 TI - Role of protein kinase C in intestinal ischemic preconditioning. AB - INTRODUCTION: Tissue protection by ischemic preconditioning (IPC) has been previously characterized in organs such as the heart and involves at least in part PKC activation. It is not yet clear whether such preconditioning against ischemia/reperfusion (I/R) injury operates in the intestine, and, if so, whether IPC involves protein kinase C (PKC). MATERIALS AND METHODS: IPC of the small intestine in male Sprague Dawley rats was induced by 10-min superior mesenteric artery (SMA) clamp followed by 120-min reperfusion. Sham-operated control or IPC rats were then rechallenged with 20-min SMA clamp. Histological injury to jejunal mucosa was assessed by microscopic examination and Parks' injury score (Grade 0 4; 0 = no damage). PKC activity was determined by immunoprecipitation of specific isoforms followed by in vitro kinase assay using mucosal scrapings of the harvested jejunum. Data were expressed as mean +/- SEM and analyzed by one-way ANOVA with multiple comparison tests. RESULTS: Ten-minute SMA clamp led to epithelial damage that was fully reversed by 120-min reperfusion. Activity of several PKC isoforms (PKCalpha, -delta, -epsilon) increased after 10-min ischemia. Epithelial injury associated with 20-min SMA clamp was attenuated by prior IPC. The protective effect of IPC on intestinal mucosa was prevented when animals were pretreated with the conventional (c) and novel (n) PKC inhibitor Go6850, but not with Go6976 (selective cPKC inhibitor), rottlerin (selective PKCdelta inhibitor), or saline control. CONCLUSIONS: Brief mesenteric ischemia induces a reversible epithelial injury in rats associated with activation of several PKC isoforms. Injury induced by mesenteric ischemia is reduced by brief ischemic preconditioning, an effect that is abolished by nonselective PKC inhibition but not by a selective inhibitor of cPKC or PKCdelta. The results suggest that activation of nPKC isoform(s), especially PKCepsilon during and following ischemic insults (IPC), may play an important role in protection against I/R injury in the intestine. PMID- 15820261 TI - Patency and structural changes in cryopreserved arterial grafts used as vessel substitutes in the rat. AB - OBJECTIVE: To evaluate the patency and structural changes that occur in the short and mid-term when cryopreserved syngenic arterial grafts are implanted in an experimental animal model. MATERIAL AND METHODS: Segments of iliac artery from the Spraque-Dawley rat were cryopreserved in a biological freezer according a controlled, computerized freezing protocol whereby the specimens are cooled at a rate of 1 degrees C/min. After storage at -145 degrees C in liquid N2 vapor for 30 days, the cryografts were slowly thawed. These vessels were grafted to the common iliac artery in syngenic animals. The following study groups were established: group I (GI), non-implanted cryografts; group II (GII), autografts; and group III (GIII), cryoisografts. The control group (CG) was comprised of fresh iliac arteries. The animals were sacrificed 14, 30, or 90 days post surgery. At each of these follow-up times, graft specimens were morphologically evaluated by light and scanning and transmission electron microscopy and immunolabeling of endothelial cells (vWf). Cell damage attributed to the cryopreservation or grafting process was also determined. RESULTS: At the time of sacrifice, graft patency was 100% for the autografts, while 26.6% of the cryoisografts showed fully occlusive thrombosis. Among other complications, two pseudoaneurysms were detected. After cryopreservation, the grafts (GI) showed patches of endothelial denudation and good cellularity of the medial layer. The intimal hyperplasia observed in autografts implanted for 14 days (GII) was significantly delayed until day 30 when the graft was cryopreserved (GIII). Cryoisografts showed general thinning of the arterial wall and degeneration accompanied by medial layer cell loss. These grafts showed most cell damage at 90 days post-implant. Expression of the vWf in all specimens showing intimal hyperplasia was confined to the outermost graft layer. CONCLUSIONS: Cryopreservation modified the reparative response of the grafts. Owing to faster degeneration of the medial layer and a delay in the appearance of intimal hyperplasia, arterial wall thickness was reduced relative to that of the non cryopreserved autografts. This thinning, at least in the short-term (90 days), does not seem to give rise to aneurysms owing to the generation of a neointima that stabilizes the vessel wall. PMID- 15820262 TI - Comparison of the ovine and porcine animal models for biocompatibility testing of vascular prostheses. AB - OBJECTIVE: Evaluation of the pig and sheep models for biocompatibility investigations of vascular prostheses (VP). DESIGN: Comparative analysis of animal experimental investigations involving two different animal models. MATERIALS AND METHODS: Commercially available polyester vascular prostheses (PET VP) were implanted into two different animal models (infrarenal porcine aorta and ovine carotid artery). The costs, surgical handling, patency rate, and healing on the basis of macroscopic, microscopic, and immunohistochemical criteria were analyzed over a period of 3 months. RESULTS: Handling and operating times (63 +/- 10 versus 76 +/- 16 min; P = 0.125) did not differ significantly. The cost of the two animal models was comparable. Integration of the VP was complete in the sheep model, but varied in the pig model (two complete, four incomplete). Complete endothelialization of all VPs was observed in the pig, which contrasted with the sheep with complete (circular) endothelialization only in the region of the anastomosis. The thickness of neointima in the region of the anastomosis differed insignificantly; immunohistochemically, only periprosthetic Ki67 was significantly reduced (28.7 +/- 9.9 versus 6 +/- 0.9%; P = 0.002) in the sheep. CONCLUSIONS: In the porcine model, extremely good endothelialization of the VP was observed, with formation of a rapid neointimal hyperplasia. The ovine model was characterized by the fact that postoperative follow-up investigations were easy to perform. Complete endothelialization was not observed. PMID- 15820263 TI - Sonoporation with doxorubicin enhances suppression of intimal hyperplasia in a vein graft model. AB - BACKGROUND: The purpose of the present study is to examine whether sonoporation with doxorubicin enhances suppression of intimal hyperplasia (IH) in a vein graft model. MATERIALS AND METHODS: After the administration of 1.5 mg/kg doxorubicin intravenously, the right external jugular vein of six rabbits was exposed at 2 W/cm2 and 1 MHz of ultrasound for 2 min (Sonoporation group). Tissue doxorubicin concentration was measured. In 48 rabbits, the right common carotid artery was ligated after performing a vein graft bypass. The animals were divided into the following four groups: the C0 group (surgical procedure only); the C0S (sonoporation without doxorubicin); the C1 (doxorubicin administration only); the C1S (sonoporation with doxorubicin). Twenty-four grafts were subjected to Elastic van Gieson staining for morphometric analysis 4 weeks after the operation; others were subjected to TdT-mediated X-dUTP nick end-labeling for detection of apoptic cells and to staining with a monoclonal antibody against the proliferating cell nuclear antigen for assessment of cell proliferation 1 week after. RESULTS: The tissue doxorubicin concentration was significantly higher in the Sonoporation group than in the Control group. Compared with the C0 group, IH was not suppressed in the C1 group but was significantly suppressed in the C1S group. Sonoporation with doxorubicin administration suppressed IH significantly (C1 group versusC1S group: P < 0.05). Cell apoptosis was induced and cell proliferation was suppressed significantly in the C1S group. CONCLUSIONS: Sonoporation with doxorubicin suppressed IH of the vein graft. Sonoporation may be effective in coronary or peripheral revascularization using vein grafts. PMID- 15820264 TI - Temporal changes in the management of diverticulitis. AB - PURPOSE: This study was designed to evaluate temporal trends in the use and type of operative and non-operative interventions in the management of diverticulitis. METHODS: A retrospective cohort using a statewide administrative database was used to identify all patients hospitalized for diverticulitis in the state of Washington (1987-2001). Poisson and logistic regression were used to calculate changes in the frequency of hospitalization, operative and percutaneous interventions, and colostomy over time. RESULTS: Of the 25,058 patients hospitalized non-electively with diverticulitis (mean age 69 +/- 16, 60% female) there were only minimal changes in the frequency of admissions over time (0.006% increase per year-IRR 1.00006 95% CI 1.00004, 1.00008). The odds of an emergency colectomy at initial hospitalization decreased by 2% each year (OR 0.98 95% CI 0.98, 0.99) whereas the odds of percutaneous abscess drainage increased 7% per year (OR 1.07 95% CI 1.05, 1.1). Among patients undergoing percutaneous drainage, the odds of operative interventions decreased by 9% compared to patients who did not have a percutaneous intervention (OR 0.91 95% CI 0.87, 0.94). The proportion of patients undergoing colostomy during emergency operations remained essentially stable over time (range 49-61%), as did the proportion of patients undergoing prophylactic colectomy after initial non-surgical management (approximately 10%). CONCLUSIONS: There was a minimal increase in the frequency of diverticulitis admissions over time. A rise in percutaneous drainage procedures was associated with a decrease in emergency operative interventions. The proportion of patients undergoing colostomy remained stable, and there does not seem to be a significant increase in the use of one-stage procedures for diverticulitis. PMID- 15820265 TI - Treatment guidelines for bipolar disorder: a critical review. AB - INTRODUCTION: The development of treatment guidelines emerged as an important element so as to standardize treatment and to provide clinicians with algorithms, which would be able to carry research findings to the everyday clinical practice. MATERIAL AND METHOD: The MEDLINE was searched with the combination of each one of the key words 'mania', 'manic', 'bipolar', 'manic-depression', 'manic-depressive' with 'treatment guidelines'. RESULTS: The search was updated until March 1st, 2004 and returned 224 articles. Twenty-seven papers concerning the publication of treatment algorithms were traced. DISCUSSION: Despite supposedly being evidence based, guidelines for the treatment of bipolar disorder vary significantly across committees or working groups. Overall, however, at the first stage of the mania/hypomania algorithm, monotherapy with lithium, divalproex sodium or olanzapine is generally recommended. At latter stages combination therapy is strongly recommended. It is clearly stated that in bipolar depression antidepressants should be used only in combination with antimanic agents in order to avoid switching of phases. During the maintenance phase all patients should receive antimanic agents, while some may need the addition of antidepressants. The most recent guidelines emphasize the use of atypical antipsychotics for mania and lamotrigine for depression. The main problem with guidelines is that they are rapidly outdated and that the evidence base relies mainly on registration monotherapy trials that hardly reflect treatment in routine clinical conditions. CONCLUSION: Treatment guidelines may be useful to avoid non-evidence-based treatment decisions, but they are quickly out-of-date and may not fully apply to the clinical setting. The more recent guidelines point the value of atypical antipsychotics, lithium, and valproate in the treatment of mania; the role of lithium, lamotrigine, and olanzapine as options for maintenance therapy; and the scarcity of options for the treatment of bipolar depression. Psychoeducation is also supported by most guidelines as an adjunctive treatment. PMID- 15820267 TI - Seasonality in a community sample of bipolar, unipolar and control subjects. AB - BACKGROUND: This study examined seasonality in a community sample of five diagnostic groups: normal subjects, those with non-seasonal depression (NSD), seasonal depression (SD), non-seasonal bipolar disorder (NSBD) and seasonal bipolar disorder (SBD). METHODS: Telephone interviews were conducted across the Province of Ontario. Seasonal changes in mood and behaviour were determined using the Seasonal Pattern Assessment Questionnaire (SPAQ). Five additional seasonality items consisting of depressive symptoms were included in the interview. The mean global severity of seasonality (GSS) scores were obtained and the entire inventory of 11 seasonality items were compared across the identified groups. RESULTS: The mean GSS score for the controls was 5.2 (S.D. = 4.0), 8.0 (S.D. = 4.9) for NSD, 10.5 (S.D. = 3.9) for SD, 10.5 (S.D. = 5.4) for NSBD and 13.4 (S.D. = 5.4) for SBD. These scores differed significantly (F = 61.68, df = 4, p < 0.001). For the majority of the individual items, the SBD group rated the highest degree of seasonal fluctuation, while the NSBD and SD groups had nearly identical item scores. LIMITATIONS: Limitations in this study include the relatively small number of subjects in the NSBD and SBD groups, and the inherent limitations in a telephone interview. CONCLUSIONS: Individuals with bipolar disorder experience greater seasonality than those with depression or healthy controls. Even the non seasonal bipolar group had as much seasonal fluctuation as the seasonal depression group, which has important implications for the management of bipolar illness. PMID- 15820266 TI - Do social anxiety disorder patients belong to a bipolar spectrum subgroup? AB - BACKGROUND: It has been proposed that all forms of bipolar disorder-perhaps all primary affective disorders-are best conceptualized as a spectrum of related illness, clinically overlapping but not necessarily genetically uniform illnesses. We aim to describe with retrospective methodology the demographic, clinical, and therapeutic response in a group of social anxiety disorder (SA) patients who improves while taking antidepressants and compare them with bipolar II (B-II) patients. METHODS: 57 SA outpatients (DSM-IV) were diagnosed and naturalistic efficacious treated with selective serotonin reuptake inhibitors (SSRI). Their demographic, clinical features and therapeutic response were compared with 41 DSM-IV bipolar II patients in their starting evaluations in our outpatient clinic in the Federal University of Rio de Janeiro, Brazil. RESULTS: There is a sub-group of SA patients who improves while taking antidepressants and presents a clear hypomanic phase. Their improvement is identical to a mild/moderate hypomanic state. Without the antidepressant, the symptoms of SA return. The SA and B-II patients have a similar number of previous depressive episodes, alcohol abuse, suicide attempts, and family history for mood disorder. LIMITATIONS: It is a retrospective data description based on a naturalist follow up. CONCLUSION: Some SA patients have demographic, clinical and therapeutic features similar to B-II patients and they might just be a Bipolar-III sub-group with a higher level of complains to social situations and without spontaneous hypomania during lifetime. PMID- 15820268 TI - Is low mood an adaptation? Evidence for subtypes with symptoms that match precipitants. AB - BACKGROUND: Although severe depression is dysfunctional, the capacity to experience normal low mood may be useful in certain fitness-threatening situations. Moreover, if specific kinds of situations recurred often enough in the course of evolution, natural selection may have shaped partially differentiated subtypes of low mood that are parallel to the subtypes of anxiety that protect against different kinds of danger. To test this hypothesis, we examined how symptoms of low mood differ depending upon the precipitating situation, and whether these differences match expectations of symptoms useful in each kind of situation. METHOD: 337 subjects who experienced a period of low mood within the last year wrote accounts describing perceived causes of their low mood and they filled out the CES-D depression inventory. Seven symptom scales were derived from analysis of CES-D data. Independent judges blindly coded the accounts into one of six precipitant categories. RESULTS: Different untoward situations were associated with different symptoms that were predicted to be useful in those situations. Social losses (death of a loved one, romantic breakups, and social isolation) were associated with greater crying and arousal. Failure to reach a goal, stress, and winter seasons were associated with more fatigue and pessimism. DISCUSSION: These results suggest that natural selection shaped not only a generic state of low mood but also partially differentiated subtypes shaped to cope with specific situations that were associated with fitness losses among our ancestors. PMID- 15820269 TI - Dopaminergic mechanism of antidepressant action in depressed patients. AB - Clinical studies have not yet determined a common mechanism of action for antidepressant drugs, which have primary sites of action on a variety of different neurotransmitter systems. However, a large body of evidence from animal studies demonstrates that sensitisation of D2-like dopamine receptors in the mesolimbic dopamine system may represent a 'final common pathway' in antidepressant action. The present study aimed to determine whether, consistent with data from animal studies, the clinical antidepressant action of selective serotonin reuptake inhibitors (SSRIs) is reversed by acute administration of a receptor antagonist selective for D2-like receptors in the mesolimbic dopamine system. The participants were patients diagnosed with major depressive disorder (n = 8) who had been treated successfully (Hamilton Depression Scale < 10) with selective serotonin uptake inhibitors (fluoxetine, citalopram or paroxetine); and age-matched, non-depressed, untreated volunteers (n = 10). They attended a psychiatric research ward on an out-patient basis, and received double-blind acute administration of either placebo, or a low dose of the selective dopamine D2/D3 receptor antagonist sulpiride (200 mg), in a counterbalanced order. Mood and psychomotor effects were assessed using visual analogue scales and the Fawcett-Clark Pleasure Capacity Scale. Sulpiride slightly improved subjective well-being in the control group, but in the antidepressant-treated patients, sulpiride caused a substantial reinstatement of depressed mood. These data are consistent with the hypothesis that sensitisation of D2-like receptors may be central to the clinical action of SSRIs. PMID- 15820270 TI - Medical comorbidity and health-related quality of life in bipolar disorder across the adult age span. AB - BACKGROUND: Little is known about medical comorbidity or health-related quality of life (HRQOL) in bipolar disorder across the adult age span, especially in public sector patients. METHODS: We obtained cross-sectional demographic, clinical, and functional ratings for 330 veterans hospitalized for bipolar disorder with Mini-Mental State score > or = 27 and without active alcohol/substance intoxication or withdrawal, who had had at least 2 prior psychiatric admissions in the last 5 years. Structured medical record review identified current/lifetime comorbid medical conditions. SF-36 Physical (PCS) and Mental (MCS) Component Scores, measured physical and mental HRQOL. Univariate and multivariate analyses addressed main hypotheses that physical and mental function decrease with age with decrements due to increasing medical comorbidity. RESULTS: PCS decreased (worsened) with age; number of current comorbid medical diagnoses, but not age, explained the decline. Older individuals had higher (better) MCS, even without controlling for medical comorbidity. Multivariate analysis indicated association of MCS with age, current depressed/mixed episode, number of past-year depressive episodes, and current anxiety disorder, but not with medical comorbidity, number of past-year manic episodes, current substance disorder or lifetime comorbidities. LIMITATIONS: This cross-sectional design studied a predominantly male hospitalized sample who qualified for and consented to subsequent randomized treatment. CONCLUSIONS: Medical comorbidity is associated with lower (worse) physical HRQOL, independent of age. Surprisingly, younger rather than older subjects reported lower mental HRQOL. This appears due in part to more complex psychiatric presentations, and several mechanisms are discussed. Both results suggest that age-specific assessment and treatment may enhance HRQOL outcome. PMID- 15820271 TI - 1H magnetic resonance spectroscopy investigation of the dorsolateral prefrontal cortex in bipolar disorder patients. AB - BACKGROUND: Magnetic resonance spectroscopy studies (MRS) reported abnormally low levels of N-acetylaspartate (NAA, a marker of neuronal integrity) in dorsolateral prefrontal cortex (DLPFC) of adult bipolar patients, suggesting possible neuronal dysfunction. Furthermore, recent MRS reports suggested possible lithium-induced increase in NAA levels in bipolar patients. We examined with in vivo (1)H MRS NAA levels in the DLPFC of adult bipolar patients. METHODS: Ten DSM-IV bipolar disorder patients (6 lithium-treated, 4 drug-free) and 32 healthy controls underwent a short echo-time 1H MRS session, which localized an 8 cm3 single-voxel in the left DLPFC using a STEAM sequence. RESULTS: No significant differences between the two groups were found for NAA, choline-containing molecules (GPC+PC), or phosphocreatine plus creatine (PCr+Cr) (Student t-test, p > 0.05). Nonetheless, NAA/PCr+Cr ratios were significantly increased in lithium-treated bipolar subjects compared to unmedicated patients and healthy controls (Mann Whitney U-test, p < 0.05). LIMITATIONS: Relatively small sample size may have reduced the statistical power of our analyses and the utilization of a single voxel approach did not allow for the examination of other cortical brain areas. CONCLUSIONS: This study did not find abnormally reduced levels of NAA in left DLPFC of adult bipolar patients, in a sample of patients who were mostly on medications. However, elevated NAA/PCr+Cr ratios were shown in lithium-treated bipolar patients. Longitudinal 1H MRS studies should further examine NAA levels in prefrontal cortex regions in untreated bipolar patients before and after mood stabilizing treatment. PMID- 15820272 TI - Lamotrigine for the treatment of bipolar spectrum disorder: a chart review. AB - BACKGROUND: Bipolar spectrum disorders are highly prevalent and disabling but are often misdiagnosed and inadequately treated. The use of mood stabilizers may be an important treatment option. This study evaluated the effectiveness and safety of lamotrigine in bipolar spectrum patients. METHODS: Retrospective follow-up of 34 charts of bipolar outpatients treated with lamotrigine in monotherapy or as add-on therapy. Clinic notes were used to score the Clinical Global Impression for Bipolar Disorders (CGI-BP-M) scale and to identify manic, hypomanic mixed or depressive relapses. RESULTS: A mean dose of lamotrigine of 113.2+/-66.6 mg/day for a mean duration of 29.7+/-17.3 weeks (range 6-96 weeks) achieved remission on depression (score 1 on the CGI-BP-M depression subscale) in 47.1% of the patients. Eleven patients (32.4%) remained euthymic (score 1 on the CGI-BP-M overall subscale) during the follow-up, after initiating treatment with lamotrigine. Effectiveness of lamotrigine was greater for those patients diagnosed with bipolar spectrum disorders other than bipolar I. Lamotrigine was well-tolerated and only two patients discontinued due to side effects. LIMITATIONS: Chart review is open to several biases. CONCLUSIONS: In this sample of bipolar spectrum patients, lamotrigine was a safe and effective treatment. PMID- 15820273 TI - Serum lipid profiles in major depression with clinical subtypes, suicide attempts and episodes. AB - BACKGROUND: We tried to report the data between serum lipid profiles and major depression with different clinical subtypes, with or without suicide attempts, and with single episode or recurrent episodes. METHOD: During a 2-year period, a total of 168 participants (109 patients with major depression and 59 subjects in a healthy control group) were recruited in this study. Blood samples for serum lipid profiles in all participants were collected. Data analysis was performed by using an analysis of covariance (ANCOVA) with body mass index (BMI) adjustment. RESULTS: The data showed that there were no significant differences of any kind in serum lipid profiles between depressive patients with melancholic feature or atypical feature, with or without suicide attempts nor between depressive patients with single episode or recurrent episodes. CONCLUSION: The results suggested there were no serum lipid profiles used as biological markers to distinguish the clinical subtypes, suicide attempts and episodes in patients with major depression. PMID- 15820274 TI - Course profiles of early- and adult-onset depression. AB - BACKGROUND: The age of onset of major depression may produce divergent profiles of recovery after treatment. METHODS: Symptom level and course trajectories of with early- and adult-onset of major depression were followed prospectively over 2 years in 62 adult outpatients who recovered from DSM-III-R unipolar major depression after cognitive behavior therapy. We compared individuals with depression onset before age 20 (n = 31) with those with onset after age 20 (n = 31), controlling for current age, education, and history of depression, and accounting for their return to mental health treatment. Participants completed psychiatric and psychosocial assessments before and after treatment, and every 6 months for 2 years post-treatment. We analyzed severity and changes of weekly depressive symptoms to construct profiles of recovery for the early- and adult onset groups. RESULTS: Our data indicated: (a) on average, the early-onset group experienced consistently more depressive symptoms than the adult-onset group; (b) the pattern of symptoms remained level for both groups during the 2-year follow up; (c) both groups maintained parallel and stable course profiles over the 2 year follow-up; and, (d) early age of onset was associated with higher rate of depressive relapse. LIMITATIONS: Accuracy of the ratings of depressive symptoms may have been limited by the retrospective nature of the psychiatric status ratings. CONCLUSIONS: Results suggest that among our recovered participants, early age of onset is associated with shorter time to relapse, more residual symptoms over the follow-up, and a stable and linear course of residual symptoms during the 2-year follow-up. PMID- 15820275 TI - The validity of the Beck Depression Inventory-Short Form as a screening and diagnostic instrument for moderate and severe depression in medical inpatients. AB - OBJECTIVE: To examine the concurrent validity of the Beck Depression Inventory Short Form (BDI-SF) to detect moderate and severe depressive episodes according to the International Classification of Diseases, 10th edition (ICD-10) criteria in inpatients with heterogeneous medical conditions and to set cut-off scores for its use in medical wards. METHODS: One hundred and fifty-five patients [53% female; mean age (+/- S.D.) = 49.5 (+/- 17) years; mean number of years of education (+/- S.D.) = 6 (+/- 4) years] consecutively admitted to the adult medical wards in a General Hospital were interviewed during the first 72 h of hospitalization. The Clinical Interview Schedule [CIS] was used to make ICD-10 psychiatric diagnoses. All patients completed the BDI-SF. A "receiver operating characteristics" (ROC) curve was obtained and the sensitivity, specificity, positive and negative predictive values were calculated for different cut-off points of the BDI-SF. RESULTS: High sensitivity and negative predictive value (NPV) were obtained with a cut-off score of 9/10 (sensitivity = 100%, specificity = 83.1%, NPV = 100%). High sensitivity and positive predictive value (PPV) were obtained with a cut-off score of 13/14 (sensitivity = 93.5%, specificity = 96%, PPV = 85.3%). The area under the ROC curve was 98.4% (95% Confidence Interval = 0.97-1.00). CONCLUSIONS: The BDI-SF is a valid instrument for detecting moderate and severe depression in medical inpatients. For screening purposes, a 9/10 cut off score is indicated, but if a high specificity is desired, a 13/14 cut-off score is warranted. PMID- 15820276 TI - Major depressive disorder in Latin America: the relationship between depression severity, painful somatic symptoms, and quality of life. AB - BACKGROUND: We report on two multi-center, prospective, observational studies (H6U-BC-LRAG and H6U-BL-LRAH) to determine the clinical profile of Latin American outpatients with major depressive disorder (MDD) and the relationship between depression severity, painful somatic symptoms, and quality of life. METHOD: Patients (n = 989) with MDD were classified according to the presence (SS+) or absence (SS-) of painful somatic symptoms using the Somatic Symptom Inventory (SSI). Visual Analogue Scale (VAS) quantified pain severity, HAMD17 and CGI-S determined depression severity, while the Quality of Life in Depression Scale (QLDS) quantified subjective well-being. RESULTS: At baseline, patients had an average CGI score of 4.5 (+/- 0.8) and HAMD17 score of 24.9 (+/- 7.2). Of the patients studied, 72.6% reported painful somatic symptoms (95% CI: 69.8, 75.4), with women 2.7 times more likely to be SS+ than men (p < 0.0001). Adjusted mean HAMD17 (26.79) and CGI-S (4.53) scores for SS+ patients were significantly (p < 0.0001) higher than for SS- patients (HAMD(17): 22.87; CGI-S: 4.28). SS+ patients had greater severity of pain across all VAS measures (p < 0.0001). The presence of somatic symptoms had a significantly deleterious effect on quality of life (p < 0.0001). CONCLUSION: Greater severity of painful somatic symptoms was associated with increased depression severity and reduced quality of life. We concluded that both emotional and physical manifestations of MDD must be addressed for successful treatment. PMID- 15820277 TI - Treatment of residual anxiety symptoms with adjunctive aripiprazole in depressed patients taking selective serotonin reuptake inhibitors. AB - BACKGROUND: Depression and anxiety are common disorders and have substantially overlapping symptom complexes. Not surprisingly, treatment approaches are similar for both conditions with the selective serotonin reuptake inhibitors (SSRIs) as the initial therapy of choice. However, after first line treatments have been deployed, residual symptoms are often problematic. Augmentation strategies to address these difficulties are an area of active investigation. This study assessed aripiprazole as adjunctive therapy to SSRIs for patients with persistent anxiety symptoms complicating a depression or anxiety disorder. METHODS: Ten patients who had been receiving SSRIs for at least 6 weeks, but still had clinically significant anxiety symptoms, were enrolled in an open label, flexibly dosed study of adjunctive aripiprazole. Clinical status was assessed with the Hamilton Anxiety Rating Scale (HAM-A), Montgomery Asberg Rating Scale (MADRS), and Sheehan Disability Scale (SDS). RESULTS: Eighty percent of the subjects had a greater than 50% reduction of symptoms on these outcome measures by week 2 of therapy, and continued with further decrements in symptoms throughout the course of the study. CONCLUSIONS: The results of this trial provide preliminary evidence that aripiprazole may be an effective adjunctive treatment in individuals on SSRIs with residual symptoms of anxiety or depression. More rigorous double-blind studies are warranted to confirm and elucidate the potential role of aripiprazole in these conditions. PMID- 15820278 TI - Advances in the non-drug, non-surgical, non-device management of chronic heart failure. AB - There have been many articles, reviews and editorials about the recent advances in pharmaceutical and device management of chronic heart failure in this and other journals over the last few years. What has been less praised are the significant advances we have made in understanding the best management of heart failure using other non-drug, non-surgical, non-device approaches. Approaches as diverse as nutrition, education, exercise, physiotherapy, psychotherapy and therapies for sleep-disordered breathing have shown considerable promise in improving the lot of our chronic heart failure (CHF) patients. Chronic heart failure is a common condition with a poor prognosis. It generates many debilitating symptoms for the sufferer. Non-pharmacologic treatment modalities play an important role alongside effective modern pharmaceutical, surgical and device therapies in relieving symptoms and improving prognosis. These treatments include those lifestyle measures that reduce the risk of underlying diseases such as coronary artery disease, diabetes, and hypertension lifestyle interventions of benefit in established CHF. Recent advances are reviewed including specialist nursing care, multi-disciplinary heart failure clinics, exercise rehabilitation, the treatment of sleep-disordered breathing, depression, obesity and cachexia. The day of the multi-disciplinary patient-centred CHF clinic has arrived and all sufferers deserve experienced management using all these approaches. PMID- 15820279 TI - Chronic heart failure patients with restrictive LV filling pattern have significantly less benefit from cardiac resynchronization therapy than patients with late LV filling pattern. AB - BACKGROUND: Cardiac resynchronization fails to improve symptoms in up to one third of patients meeting criteria for this treatment, for reasons which are unclear. Indeed, the very mechanism of benefit from resynchronization is controversial. Resynchronization may work by improving ventricular filling: we tested the hypothesis that benefit from resynchronization depends on filling pattern. METHODS AND RESULTS: We assessed symptoms (NYHA class) and LV filling of 40 patients with chronic heart failure and prolonged QRS who underwent resynchronization. Fifteen had restrictive filling pattern (E velocity>or=1.0 m/s, E/A ratio>1 and E wave deceleration time0.35. In patients with congestive heart failure, atrial fibrillation/atrial flutter was associated with an increased in-hospital mortality (OR=1.5 (1.2-1.9); p<0.001) and increased 30 day mortality (OR=1.4 (1.1-1.7); p<0.001) but not in patients without congestive heart failure. In hospital survivors, atrial fibrillation/atrial flutter was associated with an increased long-term mortality in all subgroups except those with left ventricular ejection fraction<0.25. CONCLUSIONS: Atrial fibrillation/atrial flutter is primarily associated with increased in-hospital mortality in heart failure patients. Long-term mortality is increased in all subgroups except those with left ventricular ejection fraction<25%. PMID- 15820288 TI - Analysis of bleeding complications associated with glycoprotein IIb/IIIa receptors blockade in patients with high-risk acute coronary syndromes: insights from the PRISM-PLUS study. AB - We aim to characterize the hemorrhagic complications and predictors of increased bleeding risk in a population of patients with high-risk acute coronary syndromes (ACS), enrolled in the PRISM-PLUS study. Patients treated with heparin plus tirofiban had more bleeding events compared to patients treated with heparin alone. No significant increase in major bleeding, thrombocytopenia, blood loss and blood products transfusions was observed among the patients who received the combination therapy. Several clinical variables were independently associated with increased risk of bleeding for both treatment groups: advanced age, lower body weight, female gender, decreased creatinine clearance (<30 ml/min). Females, patients with impaired renal function, patients requiring percutaneous coronary intervention (PCI), especially prolonged PCI (>100 min duration) or coronary artery bypass surgery (CABG) were at risk for increased major bleeding complications. Increased blood loss was also found in females, patients with elevated diastolic blood pressure, PCI, duration of PCI>100 min or CABG. No incremental risk was detected with the addition of tirofiban to heparin in patients at risk for major bleeding or increased blood loss. We concluded that identification of patients with high-risk ACS, at risk for bleeding complications and blood loss can be done with specific clinical variables. Tirofiban added to heparin increased minor hemorrhagic complications. Although there was no significant increase in major bleeding, thrombocytopenia and blood transfusions with the combination of tirofiban plus heparin, the power to detect a statistically significant difference in these endpoints was limited by the small number of events. PMID- 15820289 TI - Diabetic patients with acute coronary syndromes in the UK: high risk and under treated. Results from the prospective registry of acute ischaemic syndromes in the UK (PRAIS-UK). AB - OBJECTIVES: Short-term randomised trials suggest that patients with diabetes mellitus (DM), admitted with acute coronary syndromes (ACS) are at increased risk of subsequent adverse events. We tested whether this hypothesis was true for an unselected population of ACS patients with and without DM admitted with non-ST elevation MI or unstable angina, in a non-trial setting over a longer term of follow-up. METHODS: Prospective, centrally, coordinated multicenter registry involving 56 centers throughout the UK (half having angiographic facilities). Consecutive patients admitted with ACS without ST elevation on the presenting ECG were followed up to 6 months. A sub-group of patients were flagged with the UK Office for National Statistics and followed-up for death over 4 years. RESULTS: Data were collected on 1046 ACS patients of whom 170 (16%) had a prior diagnosis of DM. DM patients had higher baseline co-morbidities and unadjusted mortality rates at 6 months (11.8% vs. 6.4%, p=0.01). After correcting for clinical variables such as age, gender, smoking status and chest pain/ischaemic ECG changes on admission, prior history of any of myocardial infarction, heart failure, hypertension, hypercholesterolemia (on treatment), stroke or coronary revascularisation (PTCA or CABG), mortality rates for DM patients were no longer significantly raised (hazard ratio 1.35, 95% CI: 0.79-2.30; p=0.27 at 6 months and 1.15, 95% CI 0.72-1.83 at 4 years). 30% of diabetics were dead after 4 years of follow-up. Patients with DM were more likely to have been revascularised at 6 months and were more likely to receive ACE inhibitors. Based on the rate of recruitment and the population covered in the study, about 21,000 patients with DM will be admitted with non-ST elevation ACS each year in the UK. CONCLUSIONS: DM is common amongst patients admitted with ACS without ST elevation and is associated with significant morbidity and mortality: approximately 1 in 8 will not survive up to 6 months and 1 in 3 to 4 years. DM patients should be managed aggressively to reduce their risk of future complications. PMID- 15820290 TI - Long-term predictors of survival for the Seven Countries Study cohort from Crete: from 1960 to 2000. AB - BACKGROUND: In 1960, all male inhabitants of a series of villages in rural Crete, born between 1900 and 1919, were invited to participate in the Seven Countries Study. Analysis of 25-year mortality data from the 16 cohorts of participants indicated that the cohort from Crete had the lowest age-standardised all-cause and coronary heart disease death rates. METHODS: At baseline, 686 Cretan men (98% of those invited) participated in health examinations. Mortality data were collected over 40 years. Time-fixed and updated covariate survival analysis techniques were applied to assess eight cardiovascular disease risk factors as long-term predictors of all-cause and cardiovascular disease mortality. RESULTS: The median survival time was 32 years. All-cause and cardiovascular mortality rates were 26 and 11 per 1000 person-years, respectively. Age (relative risk 1.11, 95% CI 1.09-1.13), diastolic blood pressure (relative risk 1.02, 95% CI 1.01-1.03), and smoking (relative risk 1.37, 95% CI 1.14-1.64) were positively associated and forced expiratory volume (relative risk 0.50, 95% CI 0.36-0.68) was negatively associated with all-cause mortality. Age (relative risk 1.13, 95% CI 1.09-1.16), diastolic blood pressure (relative risk 1.01, 95% CI 1.001-1.03), and forced expiratory volume (relative risk 0.53, 95% CI 0.32-0.89) were independent predictors of cardiovascular mortality. Serum cholesterol concentration and body mass index were not independently associated with death risk. CONCLUSIONS: The Cretan cohort displays favourable 40-year survival. Even so, long-term predictors of the hazard of both all-cause and cardiovascular disease mortality are present. PMID- 15820291 TI - Exercise training modulates cytokines activity in coronary heart disease patients. AB - BACKGROUND: Physical activity may lower the risk for coronary artery disease (CAD) by mitigating inflammation, which plays a key role in the pathophysiology of atherosclerosis. The purpose of this study was to determine the effect of aerobic exercise training on levels of pro- and anti-inflammatory cytokines, IL 1, IL-6, IL-10, INF-gamma, and C-reactive protein (CRP), in CAD patients participating in a cardiac rehabilitation program. METHODS AND RESULTS: Twenty eight patients, age 64+/-7.1 years, participated in a 12-week aerobic exercise training program at 70-80% of individual maximal heart rate. Training resulted in a significant reduction of all pro-inflammatory cytokines, CRP from 7.5+/-4.2 to 3.9+/-3.5 mg/l, p<0.001, IL-1, 0.33+/-0.23 to 0.51+/-0.12 pg/ml, p=0.014, IL-6, 2.50+/-1.50 to 1.44+/-0.57 pg/ml, p=0.002, INF-gamma, 18.63+/-3.31 to 16.77+/ 2.49 pg/ml, p<0.001, as well as a significant increase in the anti-inflammatory, cytokine IL-10, from 1.61+/-1.40 to 2.29+/-2.01 pg/ml, p=0.008. Baseline CRP levels were 36% (p=0.006) higher among diabetes mellitus patients and training was associated with a 40.5% CRP reduction in these patients compared to 19% reduction in non-diabetics, p<0.01. At baseline 72% of patients were in a high risk category (CRP>3 mg/l), 28% in an intermediate (CRP=1-3 mg/l), with none in a low risk category (<1 mg/l). Following exercise training, 11% were in the low risk, 50% in the intermediate and 39% in the high risk category, indicating 46% reduction in the number of subjects in the high risk category. CONCLUSIONS: Aerobic exercise training in CAD patients is an effective mean in inducing reduction in CRP, IL-1, IL-6, INF-gamma levels, and increase in IL-10, thus, possibly improving coronary risk profile. PMID- 15820292 TI - Treatment and outcomes of eastern Europeans with coronary syndromes in OPUS-TIMI 16. AB - BACKGROUND: Registries and clinical trials have offered limited evidence on the translation of acute coronary syndrome trial findings into local practices in Eastern Europe. We examined differences in acute coronary syndrome treatment and outcomes between Eastern Europe and other regions in OPUS TIMI-16, a multinational trial of oral glycoprotein IIb/IIIa inhibition for ST-segment and non-ST-segment elevation acute coronary syndromes conducted between 1997 and 1999. METHODS AND RESULTS: We compared variation in baseline characteristics, adjunctive treatment, and outcomes of patients in Eastern Europe and other regions of the world. We found that during this period, Eastern European sites made significantly more use of medications for coronary prevention before and after hospitalization. These sites, however, also made less use of adjunctive medications during their hospitalization. Fewer Eastern European patients received lipid-lowering medication both prior to admission and following discharge. Eastern European patients also underwent fewer percutaneous coronary interventions. Patients in Eastern Europe had worse outcomes at 30 days and at 10 months. CONCLUSION: This study revealed disparities in the use of medical treatments and percutaneous coronary interventions between Eastern European sites and trial sites in other countries. These findings indicate the need for studies to address the sources of treatment and outcome variation. The underuse of effective treatment modalities in all regions calls for strategies to improve access to cardiovascular therapies. PMID- 15820293 TI - Cigarette smoking and acute coronary syndromes: a multinational observational study. AB - PURPOSE: To determine the impact of cigarette smoking on the presentation, treatment, and in-hospital outcomes of patients admitted with the full spectrum of acute coronary syndromes. METHODS: GRACE is a multinational observational registry involving 94 hospitals in 14 countries. This analysis is based on 19,325 patients aged at least 18 years admitted for acute coronary syndromes as a presumptive diagnosis with at least one of the following: electrocardiographic changes consistent with acute coronary syndromes, serial increases in serum biochemical markers of cardiac necrosis, and/or documentation of coronary artery disease. The main outcomes measured were mode of presentation, treatment and in hospital death in the ST-segment elevation myocardial infarction, non-ST-segment elevation myocardial infarction, and unstable angina groups to assess the impact of smoking status. RESULTS: Smokers were more frequently diagnosed with ST segment elevation myocardial infarction (46.0%) than former smokers (27.4%) and non-smokers (30.2%) (P<0.001). Smokers were mostly men, were younger and more aggressively treated than former smokers and non-smokers across the three acute coronary syndrome groups. Unadjusted in-hospital mortality rates were lower in smokers compared with former smokers and non-smokers in the study population (3.3%, 4.5%, and 6.9%, respectively, P<0.001), and in the ST-segment elevation myocardial infarction and non-ST-segment elevation myocardial infarction groups. However, by multivariate logistic analysis, the adjusted in-hospital mortality rate was similar regardless of smoking status. CONCLUSIONS: There is no survival advantage related to current or prior cigarette smoking in patients admitted with acute coronary syndromes, regardless of presentation. In this large multinational registry, the smokers' paradox does not exist. PMID- 15820294 TI - Adjusted indirect comparison of intracoronary drug-eluting stents: evidence from a metaanalysis of randomized bare-metal-stent-controlled trials. AB - AIMS: Drug-eluting stents (DES) have been recently investigated, with favorable data for many devices, but comparative data are lacking. We thus performed an adjusted indirect comparison metaanalysis of DES. METHODS: Randomized trials comparing DES vs. bare-metal stents (BMS) were systematically searched, and random effect odds ratios (OR) were computed for target lesion revascularization (TLR) and binary in-stent restenosis rate (BRR) at 6-12 months. We then generated interaction OR for the comparison of different DES. RESULTS: We pooled data from 17 studies (allocating 3048 patients to BMS and 3392 to nine different DES). Indirect head-to-head comparison of sirolimus-eluting Cypher (N=1007) vs. polymeric paclitaxel-eluting Taxus (N=959) showed nonsignificant differences in TLR [OR=0.8 (0.5-1.4), p=0.45] but significant reductions in BRR favoring Cypher [OR=0.3 (0.1-0.6), p<0.001]. Everolimus-eluting stents appeared noninferior to Cypher or Taxus (p>0.50 for both TLR and BRR). Actinomycin-, mycophenolate-, and apolymeric paclitaxel-eluting stents (PES) all proved significantly worse than Cypher or Taxus for TLR or BRR. CONCLUSION: Notwithstanding its inherent limitations, the present metaanalysis confirms the effectiveness of both Cypher and Taxus, supports the promising role of everolimus-eluting stents, and suggests the significant inferiority of most other devices. These post hoc findings, albeit intriguing, await prospective confirmation. PMID- 15820295 TI - Risk of cardiovascular death in elderly patients with possible heart failure. B type natriuretic peptide (BNP) and the aminoterminal fragment of ProBNP (N terminal proBNP) as prognostic indicators in a 6-year follow-up of a primary care population. AB - Heart failure is common in the elderly population and carries a serious prognosis. We evaluated EDTA-plasma B-type natriuretic peptide (brain natriuretic peptide, BNP) and the aminoterminal fragment of proBNP (N-terminal proBNP) as prognostic markers in elderly primary care patients with symptoms of heart failure. METHODS: From 474 patients attending primary care for symptoms of dyspnea, fatigue and/or peripheral edema, blood was sampled in plastic tubes containing EDTA to measure BNP by non-extraction immunoradiometric assay and N terminal proBNP by non-extraction radioimmunoassay. Patients were evaluated with respect to history and function by NYHA classification and Doppler echocardiography. Follow-up time was 6 years. Cox regression analysis was performed to identify the weight of risk variables. CONCLUSION: Total 6-year mortality was 20% (102 patients out of 510), and cardiovascular (CV) mortality was 14% (71 patients, 70% of total mortality). BNP and N-terminal proBNP were essentially equally useful as prognostic markers. In patients with the highest quartiles of plasma concentration of BNP and N-terminal proBNP, respectively, the risk of cardiovascular mortality was 10 and 4.8 times, respectively, higher than that in those in the lowest quartile. Peptide concentrations varied widely within all functional groups including those with normal echocardiographic findings. Plasma concentrations of BNP and N-terminal proBNP give important prognostic information concerning risk of cardiovascular mortality. Cost-effective "clinical pathways" should be outlined for patients with elevated peptide concentrations. PMID- 15820296 TI - Geographical influences on the association between adherence to the Mediterranean diet and the prevalence of acute coronary syndromes, in Greece: the CARDIO2000 study. AB - OBJECTIVE: We evaluated the interaction between adherence to the Mediterranean diet and region of Greece on the likelihood of having acute coronary syndromes (ACS). METHODS: During 2000-2001, a random sample of 848 patients (61+/-10 years) with their first coronary heart disease event, and 1078 frequency matched (by age sex) controls with no cardiovascular disease in their medical history, from all the country, entered into the study. Among several factors, adherence to the Mediterranean diet was assessed by a diet-score that incorporated the inherent characteristics of this diet. RESULTS: The multi-adjusted analysis showed that a 10-unit increase in the diet score was associated with a 27% (95% CI 0.66 to 0.89) decrease of the odds of having ACS. Moreover, a highly significant interaction was observed between region and diet score (p<0.001). The odds ratios varied from roughly 0.5 in Southern to 1.2 or more in Northern Greek regions (p for heterogeneity<0.05). Differences in food patterns consumed did not explain the previous findings. In addition, when we stratified our analysis by rural and urban areas we found significant differences in the estimated odds ratios (p for interaction between diet score and area=0.01), since a 10-unit increase in the diet score was associated with 22% (95% CI 0.63 to 0.96) lower odds n urban areas and 31% (95% CI 0.48 to 0.98) lower odds in rural areas. CONCLUSION: Our findings underline the significance of the Mediterranean diet on the primary prevention of ACS. Moreover, we revealed a geographical variation in the importance of this dietary pattern on coronary risk, independent from the composition of food patterns followed and the prevalence of the common cardiovascular risk factors. PMID- 15820297 TI - Carvedilol reduces the costs of medical care in severe heart failure: an economic analysis of the COPERNICUS study applied to the United Kingdom. AB - BACKGROUND: The aim of this study was to determine the effects of carvedilol on the costs related to the treatment of severe chronic heart failure (CHF). METHODS: Costs for the treatment for heart failure within the National Health Service (NHS) in the United Kingdom (UK) were applied to resource utilisation data prospectively collected in all patients randomized into the Carvedilol Prospective Randomized Cumulative Survival (COPERNICUS) Study. Unit-specific, per diem (hospital bed day) costs were used to calculate expenditures due to hospitalizations. We also included costs of carvedilol treatment, general practitioner surgery/office visits, hospital out-patient clinic visits and nursing home care based on estimates derived from validated patterns of clinical practice in the UK. RESULTS: The estimated cost of carvedilol therapy and related ambulatory care for the 1156 patients assigned to active treatment was pound530,771 ( pound44.89 per patient/month of follow-up). However, patients assigned to carvedilol were hospitalised less often and accumulated fewer and less expensive days of admission. Consequently, the total estimated cost of hospital care was pound3.49 million in the carvedilol group compared with pound4.24 million for the 1133 patients in the placebo arm. The cost of post discharge care was also less in the carvedilol than in the placebo group ( pound479,200 vs. pound548,300). Overall, the cost per patient treated in the carvedilol group was pound3948 compared to pound4279 in the placebo group. This equated to a cost of pound385.98 vs. pound434.18, respectively, per patient/month of follow-up: an 11.1% reduction in health care costs in favour of carvedilol. CONCLUSIONS: These findings suggest that not only can carvedilol treatment increase survival and reduce hospital admissions in patients with severe CHF but that it can also cut costs in the process. PMID- 15820298 TI - Are there ethnic differences in the circadian variation in onset of acute myocardial infarction? A comparison of 3 ethnic groups in Birmingham, UK and Alicante, Spain. AB - We hypothesised that ethnicity may influence the circadian pattern in acute myocardial infarction (MI), in view of the potential differences in genetic background, cardiovascular risk factors and cultural habits. To test our hypothesis, we studied 340 consecutive acute MI patients (268 males; mean age 61.6+/-12.3 years) from two different city-centre teaching hospitals in Birmingham (United Kingdom) and Alicante (Spain). A different circadian rhythm in MI onset was observed between the ethnic groups (p=0.001), with a significantly higher number of acute MI onset occurring between midnight and noon in British Caucasians and Indo-Asians. In contrast, Mediterranean Caucasians showed the converse circadian pattern, with most of the acute MI events happened between noon and midnight. Indo-Asian patients were the youngest patient group and showed the highest prevalence of diabetes and increased body mass index. Mediterranean patients had the highest prevalence of smokers but their mean serum cholesterol was the lowest. No differences in sex, blood pressure, height and weight were observed. In conclusion, this study has shown a different circadian rhythm in acute MI onset between 3 ethnic groups from two different city-centre teaching hospitals in Birmingham (United Kingdom) and Alicante (Spain) and, for the first time, provide data in the Indo-Asian population. Further studies are required to determine the pathophysiological mechanism(s) underlying these differences. PMID- 15820299 TI - Prospective cohort study of hostility and the risk of cardiovascular disease mortality. AB - BACKGROUND: Recent literature reviews have questioned hostility as a risk factor for heart disease. However, controversy persists due to the rarity of large-scale prospective cohort studies of initially healthy populations. METHODS: We prospectively investigated the association between hostility and cardiovascular (and all-cause) mortality among 20,550 men and women, 41-80 years of age, participating in the European Prospective Investigation into Cancer and Nutrition in Norfolk (EPIC-Norfolk), United Kingdom study. Participants were recruited by post from general practice age-sex registers and subsequently attended health checks that included the assessment of coronary disease risk factors. Hostility assessment was completed by postal questionnaire. RESULTS: During mean follow-up of 6 years, 1284 deaths were recorded including 481 from cardiovascular disease (CVD). Hostility was not associated with CVD mortality, after adjustment for age and prevalent disease, in either men (rate ratio for a 1 SD decrease in hostility score, representing increased hostility, 1.09; 95% confidence interval 0.98-1.22) or in women (rate ratio 1.00; 95% confidence interval 0.86-1.26). Subgroup analysis suggested hostility may be associated with CVD mortality (independent of age, prevalent disease and cigarette smoking) for participants reporting very high hostility and for those aged less than 60 years. CONCLUSIONS: Hostility was not associated with an increased risk of cardiovascular mortality in this population study of older adults. PMID- 15820300 TI - Mechanism of seasonal variation in acute myocardial infarction. PMID- 15820302 TI - Tachycardia amongst subjects recovering from severe acute respiratory syndrome (SARS). AB - SARS is a new infection in human. Patients recovering from SARS had palpitation in the form of sinus tachycardia. This study to identify the possible causes for the tachycardia excluded active disease, thyroid dysfunction, haematological, cardiac, autonomic and significant pulmonary defect at 2 months from onset of disease. The symptomatology was attributed to physical deconditioning and anxiety state. Physical and psychological fitness should be restored with rehabilitation. PMID- 15820303 TI - Five years of progress for international journal of cardiology. PMID- 15820304 TI - Sense and antisense Foxl2 transcripts in mouse. AB - FOXL2 is a forkhead transcription factor involved in eyelid development and in the development and adult function of the ovary in mammals. In mouse, we have previously suggested the existence of two mRNA isoforms of Foxl2 that result from an alternative polyadenylation. In this study, we characterize in depth the structure and expression of these two variants. We also describe an antisense transcript that overlaps the whole Foxl2 transcription unit. This antisense transcript, called Foxl2OS (for opposite strand), yields several isoforms resulting from alternative splicing. No significant coding region was found in the Foxl2OS sequence. Foxl2OS displays a pattern of expression very similar to that of Foxl2 in the gonads during development and at the adult age. RNA FISH experiments show that both transcripts are expressed in the same cells at the same time. We suggest that Foxl2OS is a noncoding antisense RNA that may be involved in the regulation of Foxl2. All in all our results provide new insights about the organization of the murine Foxl2 locus. This might help us understand its regulation and function. PMID- 15820305 TI - Molecular characterization of the pericentric inversion of chimpanzee chromosome 11 homologous to human chromosome 9. AB - In addition to the fusion of human chromosome 2, nine pericentric inversions are the most conspicuous karyotype differences between humans and chimpanzees. In this study we identified the breakpoint regions of the pericentric inversion of chimpanzee chromosome 11 (PTR 11) homologous to human chromosome 9 (HSA 9). The break in homology between PTR 11p and HSA 9p12 maps to pericentromeric segmental duplications, whereas the breakpoint region orthologous to 9q21.33 is located in intergenic single-copy sequences. Close to the inversion breakpoint in PTR 11q, large blocks of alpha satellites are located, which indicate the presence of the centromere. Since G-banding analysis and the comparative BAC analyses performed in this study imply that the inversion breaks occurred in the region homologous to HSA 9q21.33 and 9p12, but not within the centromere, the structure of PTR 11 cannot be explained by a single pericentric inversion. In addition to this pericentric inversion of PTR 11, further events like centromere repositioning or a second smaller inversion must be assumed to explain the structure of PTR 11 compared with HSA 9. PMID- 15820306 TI - Mammalian SP/KLF transcription factors: bring in the family. AB - The advent of the genome projects has provided new avenues to explore the question of how DNA sequence information is used appropriately by mammalian cells. Regulation of transcription is not the only, but is certainly a very important, mechanism involved in this process. We can now identify all the genes encoding transcription factors belonging to a certain class and study their biological functions in unprecedented detail through the use of an array of biomolecular tools. It is important to use rigorous and uniform definitions for the classification of transcription factors, because this helps us to comprehend the functions of transcription factor families in biological networks. Here, we propose an unambiguous nomenclature for the members of the Specificity Protein/Kruppel-like Factor (SP/KLF) transcription factor family. PMID- 15820307 TI - Mouse splice mutant generation from ENU-treated ES cells--a gene-driven approach. AB - Mutant mice are important for elucidating mammalian gene functions and for modeling human disease phenotypes. In recent years, chemical mutagenesis has become an increasingly popular method to disrupt gene functions due to its high efficiency of inducing mutations throughout the genome. Mutagenesis of embryonic stem (ES) cells offers the possibility of gene-driven approaches, which, however, require efficient mutation detection procedures to screen archives of mutated samples for lesions in particular genes. We have developed an approach that focuses on the detection of splice mutations in highly pooled cDNA samples using exon-skipping PCR primers. As a proof of concept, splice mutants for the Kit gene were isolated from a library comprising approximately 40,000 ES cell clones treated with N-ethyl-N-nitrosourea followed by transmission through the mouse germ-line. The approach will be useful for the production of mouse models for human disease-related splice mutations and as a general gene disruption strategy. PMID- 15820308 TI - Linkage disequilibrium analysis identifies an FGFR1 haplotype-tag SNP associated with normal variation in craniofacial shape. AB - Mutations in FGFR1 and TWIST1 have been reported to affect the timing of calvarial suture fusion resulting in craniosynostosis and facial abnormalities. We screened nonpathologic populations for genetic polymorphisms that may associate with normal craniofacial variation. We identified 17 single-nucleotide polymorphisms (SNPs) in FGFR1, 6 of which were novel (g.8591855G-->A, g.8593685G- >A, g.8602303C-->T, g.8602475A-->G (p.Ile293Val), g.8605849C-->T, g.8607868G- >A). No SNPs were found in TWIST1. FGFR1 SNP haplotypes were reconstructed for Caucasian, Asian, Australian Aboriginal, and African American populations. All populations shared two linkage disequilibrium blocks, with one haplotype-tag SNP (htSNP) tagging each block. The htSNP g.8592931G-->C was found to have a significant negative correlation with the cephalic index for all populations (R = -0.187, p = 0.036), with larger correlations in Asians and females. This finding is a starting point in the identification of a set of SNPs that can be genotyped to determine both normal and disease craniofacial phenotypes. PMID- 15820309 TI - Distribution of human beta-defensin polymorphisms in various control and cystic fibrosis populations. AB - Human beta defensins contribute to the first line of defense against infection of the lung. Polymorphisms in these genes are therefore potential modifiers of the severity of lung disease in cystic fibrosis. Polymorphisms were sought in the human beta-defensin genes DEFB1, DEFB4, DEFB103A, and DEFB104 in healthy individuals and cystic fibrosis (CF) patients living in various European countries. DEFB1, DEFB4, and DEFB104 were very polymorphic, but DEFB103A was not. Within Europe, differences between control populations were found for some of the frequent polymorphisms in DEFB1, with significant differences between South Italian and Czech populations. Moreover, frequent polymorphisms located in DEFB4 and DEFB104 were not in Hardy Weinberg equilibrium in all populations studied, while those in DEFB1 were in Hardy Weinberg equilibrium. Sequencing of a monochromosomal chromosome 8 mouse-human hybrid cell line revealed signals for multiple alleles for some loci in DEFB4 and DEFB104, but not for DEFB1. This indicated that more than one DEFB4 and DEFB104 gene was present on this chromosome 8, in agreement with recent findings that DEFB4 and DEFB104 are part of a repeat region. Individual DEFB4 and DEFB104 PCR amplification products of various samples were cloned and sequenced. The results showed that one DNA sample could contain more than two haplotypes, indicating that the various repeats on one chromosome were not identical. Given the higher complexity found in the genomic organization of the DEFB4 and DEFB104 genes, association studies with CF lung disease severity were performed only for frequent polymorphisms located in DEFB1. No association with the age of first infection by Pseudomonas aeruginosa or with the FEV1 percentage at the age of 11-13 years could be found. PMID- 15820310 TI - The Mass1frings mutation underlies early onset hearing impairment in BUB/BnJ mice, a model for the auditory pathology of Usher syndrome IIC. AB - The human ortholog of the gene responsible for audiogenic seizure susceptibility in Frings and BUB/BnJ mice (mouse gene symbol Mass1) recently was shown to underlie Usher syndrome type IIC (USH2C). Here we report that the Mass1frings mutation is responsible for the early onset hearing impairment of BUB/BnJ mice. We found highly significant linkage of Mass1 with ABR threshold variation among mice from two backcrosses involving BUB/BnJ mice with mice of strains CAST/EiJ and MOLD/RkJ. We also show an additive effect of the Cdh23 locus in modulating the progression of hearing loss in backcross mice. Together, these two loci account for more than 70% of the total ABR threshold variation among the backcross mice at all ages. The modifying effect of the strain-specific Cdh23ahl variant may account for the hearing and audiogenic seizure differences observed between Frings and BUB/BnJ mice, which share the Mass1frings mutation. During postnatal cochlear development in BUB/BnJ mice, stereocilia bundles develop abnormally and remain immature and splayed into adulthood, corresponding with the early onset hearing impairment associated with Mass1frings. Progressive base-apex hair cell degeneration occurs at older ages, corresponding with the age-related hearing loss associated with Cdh23ahl. The molecular basis and pathophysiology of hearing loss suggest BUB/BnJ and Frings mice as models to study cellular and molecular mechanisms underlying USH2C auditory pathology. PMID- 15820311 TI - Identification of candidate genes in the type 2 diabetes modifier locus using expression QTL. AB - To identify new genetic determinants relevant to type 2 diabetes (T2D), diabetic F2 progeny were generated by intercrossing F1 mice obtained from a cross of BKS.Cg-Lepr(db)+/+m and DBA/2, and T2D-related phenotypes were measured. In the F2 population, increased susceptibility to diabetes and obesity was observed. We also detected the major quantitative trait loci (QTL) modifying the severity of diabetes on chromosome 9, where peaks of logarithm of odds (LOD) overlapped for three traits. To identify candidate genes in the QTL intervals, we combined "expression QTL" (eQTL), taking mRNA levels as quantitative traits, and "interstrain sequence variations, including cSNPs." As a result, four genes were identified from cosegregation of clinical QTL with eQTL and 13 genes were found from interstrain cSNPs as candidates in the T2D modifier QTL. Our combined approach shows the acceleration of the discovery of candidate genes in the QTL of interest, spanning several megabases. PMID- 15820312 TI - Isolation and analysis of candidate myeloid tumor suppressor genes from a commonly deleted segment of 7q22. AB - Monosomy 7 and deletions of 7q are recurring leukemia-associated cytogenetic abnormalities that correlate with adverse outcomes in children and adults. We describe a 2.52-Mb genomic DNA contig that spans a commonly deleted segment of chromosome band 7q22 identified in myeloid malignancies. This interval currently includes 14 genes, 19 predicted genes, and 5 predicted pseudogenes. We have extensively characterized the FBXL13, NAPE-PLD, and SVH genes as candidate myeloid tumor suppressors. FBXL13 encodes a novel F-box protein, SVHis a member of a gene family that contains Armadillo-like repeats, and NAPE-PLD encodes a phospholipase D-type phosphodiesterase. Analysis of a panel of leukemia specimens with monosomy 7 did not reveal mutations in these or in the candidate genes LRRC17, PRO1598, and SRPK2. This fully sequenced and annotated contig provides a resource for candidate myeloid tumor suppressor gene discovery. PMID- 15820313 TI - Defining a holoprosencephaly locus on human chromosome 14q13 and characterization of potential candidate genes. AB - Holoprosencephaly (HPE) is the most common developmental field defect in patterning of the human prosencephalon and associated craniofacial structures. The genetics is complex, with 12 loci defined on 11 chromosomes. We defined a locus for HPE (HPE8) on human chromosome 14q13 between markers D14S49 and AFM205XG5, by mapping deletion intervals of affected subjects with proximal chromosome 14q interstitial cytogenetic deletions. A 35-BAC contig was built by chromosome walking. By annotation of the 2.82-Mb minimal critical region, we identified 28 possible genes. Seven genes were expressed in human fetal brain: NPAS3, SNX6, C14ORF11, C14ORF10, PAX9, NKX2.1, and C14ORF19, the last an apparent gene fragment. Molecular embryology, animal modeling, and human mutation studies were reported elsewhere for PAX9 and NKX2.1. We focused on three genes, SNX6, NPAS3, and C14ORF11, as potential candidates for HPE. Genomic structure, human expression patterns, protein cellular localization, and embryonic expression patterns of orthologous murine genes were determined, showing that the three genes have properties similar to those of known HPE genes. PMID- 15820314 TI - A reciprocal translocation 46,XY,t(8;9)(p11.2;q13) in a bladder exstrophy patient disrupts CNTNAP3 and presents evidence of a pericentromeric duplication on chromosome 9. AB - A patient with sporadic bladder exstrophy and de novo apparently balanced chromosomal translocation 46,XY,t(8;9)(p11.2;q13) was analyzed by fluorescence in situ hybridization (FISH) and molecular methods. We were able to map both translocation breakpoints to single genomic clones. The chromosome 8p11.2 breakpoint was mapped to BAC clone RP4-547J18, predicted to contain several hypothetical genes. Characterization of the chromosome 9q13 breakpoint indicated a disruption in the 5' region of CNTNAP3 within BAC RP11-292B8. This observation suggests possible involvement of CNTNAP3 in the etiology of bladder exstrophy. Additionally, FISH analysis identified several genomic copies of CNTNAP3 on both sides of the chromosome 9 centromere flanking the polymorphic heterochromatin. Northern blot analysis of lymphoblast and bladder RNA confirmed CNTNAP3 transcripts in these tissues and did not show abnormal CNTNAP3 expression in the proband and two unrelated patients with bladder exstrophy. The identification of multiple copies of three BAC clones in the proband, his parents, and unrelated controls suggests that duplications of CNTNAP3 and the surrounding genomic region have occurred as a result of repeated events of unequal crossing over and pericentric inversions during chromosome 9 evolution. PMID- 15820315 TI - A nonimprinted Prader-Willi Syndrome (PWS)-region gene regulates a different chromosomal domain in trans but the imprinted pws loci do not alter genome-wide mRNA levels. AB - Prader-Willi syndrome (PWS) is a complex neurobehavioral disorder that results from loss of function of 10 clustered, paternally expressed genes in a 1.5-Mb region of chromosome 15q11-q13. Many of the primary PWS region genes appear to have nuclear RNA regulatory functions, suggesting that multiple genetic pathways could be secondarily affected in PWS. Using a transgenic mouse model of PWS (TgPWS) with an approximately 4-Mb chromosome 7C deletion of paternal origin that models the neonatal phenotype of the human syndrome we compared by oligonucleotide microarrays expression levels of approximately 12,000 genes and ESTs in TgPWS and wild-type brain. Hybridization data were processed with two distinct statistical algorithms and revealed a dramatically reduced expression of 4 imprinted genes within the deletion region in TgPWS mice, with 2 nonimprinted, codeleted genes reduced twofold. However, only 3 genes outside the deletion were significantly altered in TgPWS mouse brain, with approximately 1.5-fold up regulation of mRNA levels. Remarkably, these genes map to a single chromosome domain (18B3), and by quantitative RT-PCR we show that 8 genes in this domain are up-regulated in TgPWS brain. These 18B3 genes were up-regulated in an equivalent manner in Angelman syndrome mouse (TgAS) brain, which has the same deletion but of maternal origin. Therefore, the trans-regulation of the chromosome 18B3 domain is due to decreased expression of a nonimprinted gene within the TgPWS/AS mouse deletion in mouse chromosome 7C. Most surprisingly, since 48-60% of the genome was screened, it appears that the imprinted mouse PWS loci do not widely regulate mRNA levels of other genes and may regulate RNA structure. PMID- 15820316 TI - Molecular cloning and functional characterization of mouse Nxf family gene products. AB - Tap, a member of the evolutionarily conserved nuclear RNA export factor (NXF) family of proteins, has been implicated in the nuclear export of bulk poly(A)+ RNAs. cDNAs encoding the mouse NXF proteins (Tap, NXF7, NXF2, and NXF3) were prepared and the gene products were characterized in terms of their genomic organization, expression patterns, and biochemical properties. Mouse Tap was found to be ubiquitously expressed, whereas tissue- and developmental stage specific expression of mouse Nxf2, Nxf3, and Nxf7 was observed. Although mouse Tap and NXF2 bound to the phenylalanine-glycine repeat sequences of nucleoporins, NXF7 and NXF3 did not. GFP-tagged mouse Tap and NXF2 were localized predominantly in the nucleus. In contrast, GFP-tagged NXF7 and NXF3 were localized exclusively in the cytoplasm. As shown for the human counterpart, disruption of the leucine rich nuclear export signal or leptomycin B treatment abolishes the cytoplasmic localization of mouse NXF3. p15/NXT1, an essential cofactor for human Tap in the export of mRNAs, was able to bind to mouse Tap, NXF2, and NXF3, but NXF7 did not form a stable heterodimeric complex. Transient transfection experiments indicated that only mouse Tap and NXF2 enhance the nuclear export of an otherwise inefficiently exported mRNA substrate. The orthologous relationship between human and mouse Nxf genes is discussed on the basis of these data. PMID- 15820317 TI - No causative DLL4 mutations in periodic catatonia patients from 15q15 linked families. AB - Two well-supported theories of schizophrenia pathogenesis are the neurotransmitter theory and the neurodevelopmental theory, suggesting, respectively, that dysregulation of neurotransmitter signaling and abnormal brain development are causative in this disease. The strongest evidence of neurotransmitter involvement are suggestions of abnormal dopamine signaling in the prefrontal cortex and one of the strongest indications of developmental abnormalities contributing to this disease is an inverse layering of the prefrontal cortex. These two theories of schizophrenia pathogenesis can be united by their involvement of the prefrontal cortex, where structural abnormalities could lead to neurochemical abnormalities. Accordingly, any gene expressed in the prefrontal cortex of developing brains is a functional candidate for schizophrenia. We have previously reported strong linkage to 15q15 (LOD = 3. 57; P = 2.6 x 10(-5)) in a collection of German multiplex families segregating the periodic catatonia subtype of schizophrenia in a nearly Mendelian fashion. A gene within our 15q15 linkage region, DLL4, is expressed in developing forebrain and produces a NOTCH4 ligand. Variants of NOTCH4 are associated with schizophrenia, thus DLL4 is both a functional as well as a positional candidate for schizophrenia. We screened this gene for mutations in three affected individuals and two unrelated controls and found two previously unreported SNPs, one non synonymous polymorphism that changed an arganine to a histadine in Exon 7 and one synonymous polymorphism in exons. The non-synonymous SNP is a rare variant in that it was not found in 100 control chromosomes; however, it did not cosegregate with the disease in the extended family so it is not causative in this pedigree. It is unlikely that mutations in DLL4 are causative in this collection of families with linkage to 15q15. PMID- 15820318 TI - Nogo CAA 3'UTR Insertion polymorphism is not associated with Schizophrenia nor with bipolar disorder. AB - The Nogo gene maps to 2p14-p13, a region consistently associated with schizophrenia and bipolar disorder. The association of a polymorphism in Nogo was previously investigated by two groups, with divergent results. In this report, using an alternative approach, we evaluated this same polymorphism in 725 individuals, including patients with schizophrenia, bipolar disorder, normal controls and non-human primate samples. Our results indicate that the polymorphism is not associated with any of these diseases, but has a remarkably biased distribution in ethnic groups. Genotyping of primate samples, suggest that this polymorphism is a recent event in human speciation. PMID- 15820319 TI - Possible association of the MAG locus with schizophrenia in a Chinese Han cohort of family trios. AB - Neurotransmitter-based hypotheses have so far led to only moderate success in predicting new pathogenetic findings in etiology of schizophrenia. On the other hand, the more recent oligodendroglia hypotheses of this disorder have been supported by an increasing body of evidence. For example, the expression level of the myelin associated glycoprotein (MAG) gene has been shown to be significantly lower in schizophrenia patient groups compared to control groups. Such an effect might be a result of genetic variations of the MAG gene. In order to test this hypothesis, we genotyped four markers within the MAG locus in 413 trios sample of the Han Chinese using allele-specific PCR. None of the four markers revealed noticeable allelic significance. However, the four-marker and two-marker haplotypes covering components rs720309 and rs720308 were observed to be significantly associated with schizophrenia (P < 0.0001) in this study. In addition, we identified one common risk haplotype TA (rs720309-rs720308, present in 78.5% of the general population) that showed increased evidence of overtransmission from parents to affected offspring (P = 0.0001). The results demonstrated MAG might play a role in genetic susceptibility to schizophrenia. Furthermore, our finding of a possible association between the MAG locus and schizophrenia is in agreement with the hypotheses of oligodendrltic and myelination dysfunction. PMID- 15820320 TI - Genetic susceptibility to tardive dyskinesia in chronic schizophrenia subjects: III. Lack of association of CYP3A4 and CYP2D6 gene polymorphisms. AB - Tardive dyskinesia is a severe debilitating movement disorder characterized by choreoathetotic movements developing in one-fifth of the patients with schizophrenia. In this study we have investigated the significance of CYP3A4*1B and CYP2D6*4 polymorphisms in TD susceptibility among chronic schizophrenia patients (n = 335) from north India. Tardive dyskinesia was diagnosed in approximately 29% (96/335) of these patients. No significant association of either of the two SNPs with TD (CYP3A4*1B chi2 = 0. 308, df = 1, p = 0.579; CYP2D6*4 chi2 = 0.006, df = 1, p = 0.935) was observed. However a trend towards increased severity of TD in patients heterozygous for the CYP2D6*4 mutation was observed. PMID- 15820321 TI - Increased expression of glutaminase and glutamine synthetase mRNA in the thalamus in schizophrenia. AB - Numerous molecules enable the handling of glutamate that is destined for neurotransmitter release, including transporters, receptors and glutamatergic enzymes. Previous work in our lab has shown altered levels of transcript expression of excitatory amino acid transporters and a vesicular glutamate transporter in the thalamus in schizophrenia. These changes suggest that molecules that facilitate the release and reuptake of glutamate may be abnormal in schizophrenia. In this study we determined the levels of expression of phosphate activated glutaminase (PAG), which converts glutamine to glutamate, and glutamine synthetase (GS), which converts glutamate to glutamine, with the hypothesis that thalamic PAG and GS transcript expression is altered in schizophrenia. We investigated expression of PAG and GS mRNA using in situ hybridization in six different thalamic nuclei (anterior, dorsomedial, centromedial, ventral anterior, ventral and reticular) from 13 persons with schizophrenia and 8 comparison subjects and found that transcripts for PAG and GS were significantly increased in schizophrenia. Increased PAG and GS transcripts suggest enhanced glutamatergic neurotransmission in the thalamus and its efferent targets in schizophrenia. PMID- 15820322 TI - Neurological abnormalities in first-episode schizophrenia: temporal stability and clinical and outcome correlates. AB - OBJECTIVE: Neurological abnormalities in subjects with schizophrenia have been regarded as diagnostically non-specific and non-localising. This study assessed the temporal stability of neurological abnormalities in subjects with first episode schizophrenia over the course of 12 months. We also examined their relationships with psychiatric symptoms, medication effects and treatment outcome. METHOD: The sample comprised 66 largely medication-naive subjects who were treated according to a fixed protocol. We performed a factor analysis of the Neurological Evaluation Scale (NES) items, and relationships between the NES factors and various clinical and outcome measures were explored. RESULTS: Five NES factors were identified, explaining 68.4% of the variance. While the NES total scores did not change significantly over time, poor performance on motor sequencing tests was related to longer duration of untreated psychosis, and showed a tendency to improve as psychiatric symptoms resolved. The most interesting finding was that high scores on the motor sequencing factor predicted the emergence of persistent dyskinesia at 24 months (ANCOVAR F(1, 20) = 19.287, p = 0.0002). CONCLUSIONS: Two NES factors (motor sequencing and attention) are reasonably replicable across samples, and have potential relevance for the further exploration of the pathogenesis of schizophrenia, as well as possible clinical applications. PMID- 15820323 TI - A 3-year prospective study of neurological soft signs in first-episode schizophrenia. AB - Neurological soft signs are biological traits that underlie schizophrenia and are found to occur at higher levels in at-risk individuals. The expression of neurological soft signs may be modifiable during the onset of the first psychotic episode and the subsequent evolution of the illness and its treatment. This study investigates neurological soft signs in 138 patients with first-episode schizophrenia and tracks the expression of motor soft signs in the following 3 years. For the 93 patients who have completed the 3-year follow-up, we find that neurological soft signs are stable in the 3 years that follow the first psychotic episode, and that neurological soft signs are already elevated at the presentation of first-episode psychosis in medication-naive subjects. The level of neurological soft signs at clinical stabilization is lower for patients with a shorter duration of untreated psychosis. Although the quantity of neurological soft signs does not significantly change in the 3 years that follow the first episode, the relationship between neurological soft signs and negative symptoms does not become apparent until 1 year after the initial episode. A higher level of neurological soft signs is related to a lower educational level and an older age at onset, but the level of neurological soft signs does not predict the outcome in terms of relapse or occupational functioning. PMID- 15820324 TI - The effects of antipsychotic medication on factor and cluster structure of neurologic examination abnormalities in schizophrenia. AB - This study extends a previous study of the factor structure of the neurologic examination in unmedicated schizophrenia, utilizing cluster analysis and adding a medicated condition. We administered a modified version of the Neurologic Evaluation Scale (NES) on two occasions to 80 patients with schizophrenia or schizoaffective disorder, once while on antipsychotic medications and once while off medication. Data were distilled by combining right- and left-side scores, and by excluding rarely abnormal and unreliable items from the analysis. Principal components analysis yielded an intuitive four-factor solution in the unmedicated condition, but an inscrutable five-factor solution during medication. Cluster analysis revealed three groups: normal, cognitively impaired, and diffusely impaired. These results were also less interpretable with data from the medicated condition. Neurologic performance was better in the medicated than in the unmedicated condition. As is the case with other domains of symptoms and performance in schizophrenia, relationships among neurologic exam variables are altered by the presence of antipsychotic medication. PMID- 15820325 TI - Relationship of neuromotor disturbances to psychosis symptoms in first-episode neuroleptic-naive schizophrenia patients. AB - From the very inception of the modern diagnostic scheme for psychotic disorders, abnormalities in motor function have been observed in these conditions. Despite convergence from multiple areas of research supporting the notion that multiple frontal-subcortical circuits regulate motor and limbic behavior, the precise relationship between motor abnormalities and psychopathology has not been elucidated. The goals of this study were to examine the prevalence of extrapyramidal signs (EPS) in first-episode schizophrenia patients and their relationships to three psychopathological dimensions (positive psychosis syndrome, negative syndrome, and disorganization). We assessed EPS using traditional observer-based as well as quantitative instrumental measures in 39 neuroleptic-naive first-episode schizophrenia subjects. Subjects were followed for 6 months after initiating antipsychotic treatment to examine the stability of motor-limbic relationships. Four main findings emerged from this study. First, depending on the measure used the prevalence of dyskinesia prior to treatment ranged from 13% to 20%. The prevalence of parkinsonism ranged from 18% to 28%. Second, severity of dyskinesia was associated with the positive psychotic syndrome; whereas parkinsonism was associated with the positive psychosis, negative syndrome and disorganization. Third, psychopathology improved significantly across all symptom dimensions following antipsychotic treatment, while EPS remained stable. This suggests that some motor abnormalities in schizophrenia may reflect trait characteristics. Fourth, abnormalities on the pre treatment instrumental measure of parkinsonism predicted greater improvement on positive psychosis symptoms following treatment (p=0.008). Our findings support the notion that neuromotor disturbances may be a core feature of schizophrenia in a substantial proportion of patients and implicate multiple fronto-striatal circuits regulating limbic and neuromotor behavior in schizophrenia. PMID- 15820326 TI - Suicidal behavior in schizophrenia and depression: a comparison. AB - The aim of this study was to compare the frequencies and suicide attempt characteristics in patients with schizophrenia (N = 25) and major depression in absence of schizophrenia (N = 107). There were no significant differences in attempt methods, but attempters with schizophrenia reported a lower number of life events influencing the attempt (2.0 vs. 2.6), less influence of life events on the attempt (42% vs. 83%) and lower GAF scores (36 vs. 50). Schizophrenia was not associated with higher impulsivity scores but with significantly more lethal attempts (40% vs. 29%) and with a trend toward (p = 0.08) higher scores in aggressive behavior history. PMID- 15820327 TI - The relationships between symptom dimensions and dysphoria in schizophrenia. AB - Previous studies have suggested that qualitatively distinct aspects of dysphoria (anxiety and depression) are related to specific dimension of schizophrenia symptomatology. Most of these studies used simple dimensions and dysphoria models, although finer distinctions could help defining specific relationships. This study examined the relationships of distinctive aspects of depression and anxiety (both state and trait) with symptom dimensions. Forty patients with a DSM IV diagnosis of schizophrenia were assessed for symptoms (SAPS-SANS), trait and state anxiety (STAI) and depression (CDS). Symptoms ratings were summarized as dimensional scores according to a two-, three- or five-dimensional models proposed in the literature. The correlation analysis replicates previous observations that distinct aspects of dysphoria are associated with specific dimensions of schizophrenia, with the exception of disorganization. Moreover, controlling for intercorrelated variables revealed that schizophrenia and dysphoric symptoms might act in combination and/or through indirect links to contribute to illness expression. Our data further suggested that these associations may be best understood in terms of interactions between various processing biases alluded in the most recent cognitive accounts of schizophrenia symptoms. PMID- 15820328 TI - Change in the relationship between anhedonia and functional deficits over a 20 year period in individuals with schizophrenia. AB - Although early theorists suggested that deficits in emotional experience be considered a hallmark characteristic of schizophrenia, there has been limited research, and inconsistent findings, on the relationship between anhedonia and functional capacity in individuals after the onset of schizophrenia. Stronger relationships have typically been reported for chronic samples in contrast to first episode samples, although it is not clear whether this is due to selection biases that influence recruitment in these different groups, or whether results reflect a change over the course of illness. The current longitudinal study examined the relationship between physical anhedonia and functional status in a sample of 61 individuals with schizophrenia at regular intervals over a 20-year period. Subjects were recruited into the study during an index hospitalization and completed assessments at 2-, 4.5-, 7.5-, 10-, 15-, and 20-year follow-ups. Analyses indicate that the relationship between anhedonia and impairments increases over time, although mean performance on these measures is stable across this same time period. These results suggest increasing convergence of impairments in emotional, adaptive, and cognitive capacities over time, with physical anhedonia associated with poorer outcome. PMID- 15820329 TI - Factor solution of the BPRS-expanded version in schizophrenic outpatients living in five European countries. AB - RATIONALE: The expanded version of the Brief Psychiatric Rating Scale (BPRS-E) has improved the instrument's coverage and interrater reliability, but there is little knowledge on its subsyndromes. OBJECTIVES: To assess: (1) whether there are common underlying BPRS-E subscales in patients living in different countries and (2) if this is the case, whether these subscales behave the same in all populations and, if not, what are the differences over these populations. METHODS: Data are part of the EPSILON study, a collaborative project carried out in Denmark, England, Holland, Italy and Spain. A random representative sample of 404 adult patients with a ICD-10 diagnosis of schizophrenia who have been in contact with mental health services of a defined catchment area in each site were assessed. Simultaneous component analysis (SCA) was used to find component weights that optimally explain the variance of the variables in different populations simultaneously. RESULTS: Symptom severity differed significantly among the five EPSILON sites in 12 out of 24 BPRS-E items, but a common component solution could be found. It explained 48.8% of the variance and gave four well interpretable components: manic excitement/disorganization, depression/anxiety, negative and positive symptoms. Each component's internal consistency and intercomponent correlation matrix differed significantly among sites. The four components mean score differed significantly among sites for negative symptoms and depression/anxiety. CONCLUSIONS: In spite of the heterogeneity of symptom's severity in the various countries, the way symptoms cluster in schizophrenia is rather stable cross-culturally. Data demonstrate that to explore schizophrenia a third component, including mania/disorganization items, is necessary beside the positive-negative symptom dimensions. The subscales derived from these analyses can be readily used in clinical trials and epidemiological studies. PMID- 15820330 TI - An empirical conceptualization of the recovery orientation. AB - OBJECTIVE: The recovery movement is having a growing impact on policy for people with severe mental illness. The empirical literature on the recovery orientation, however, is scant, and no empirical conceptualization of recovery has been published. METHOD: We identified items reflecting recovery themes and measuring aspects of subjective experience, and used principle components and confirmatory factor analyses to develop an empirical conceptualization of the recovery orientation, using data from a large, systematic study of schizophrenia. RESULTS: We identified four domains of the recovery orientation: empowerment, hope and optimism, knowledge and life satisfaction. CONCLUSIONS: We propose here an initial approach to measuring and conceptualizing recovery attitudes. We also suggest that the evidence-based practice (EBP) movement may help to identify interventions that promote the recovery orientation and help to advance recovery attitudes. We suggest that there is a bidirectional relationship between recovery attitudes and the positive clinical outcomes that are the goals of EBPs. Through the use of empirically derived conceptualizations of recovery, EBPs can provide a mechanism for identifying treatments that promote the recovery orientation. The conceptualization proposed here can, thus, serve as a tool to assess changes in recovery attitudes during participation in specific EBPs. PMID- 15820331 TI - Needs and its relation to symptom dimensions in a sample of outpatients with schizophrenia. AB - OBJECTIVE: To analyse the association between symptom dimensions in schizophrenia and number and type of met and unmet needs. METHOD: A sample of 231 outpatients randomly selected from a register that included all patients treated in 5 mental health care centres (MHCC) was evaluated. Assessment instruments included the Camberwell Assessment of Needs (CAN) questionnaire and the PANSS. RESULTS: Number of needs are related to overall severity of psychopathology. Patients with more symptoms have more total needs (p < 0.001) and unmet needs (p < 0.001). A multiple lineal regression model showed that the disorganized and excited dimensions of the PANSS are the most important components for explaining the variance of number of needs. Type of needs is related to subtypes of schizophrenia, specially with disorganized and excited symptoms. CONCLUSION: Psychosocial needs are related to schizophrenia subtypes. Patients with more needs are those with more disorganized and excitatory symptoms. PMID- 15820332 TI - Cannabis use prior to first onset psychosis predicts spared neurocognition at 10 year follow-up. AB - A priori cannabis use was recorded at index admission for 112 participants in the Manchester first-episode psychosis cohort. 69 of the 100 surviving (mainly schizophrenia) patients were followed up 10-12 years later and assessed on a battery of clinical, behavioural and neurocognitive measures. Individuals who had not used cannabis before the first episode of illness were generally indistinguishable from cannabis users at follow-up, except that the latter group evidenced a marked 'sparing' of neurocognitive functions. These findings are briefly discussed in relation to other casual factors in psychosis. PMID- 15820333 TI - Further study of a genetic association between the CLDN5 locus and schizophrenia. PMID- 15820335 TI - Editorial. PMID- 15820334 TI - The soluble leptin receptor in olanzapine-induced weight gain: results from a prospective study. PMID- 15820336 TI - Should neurologists be trained to recognize and treat comorbid depression of neurologic disorders? Yes. AB - Depression is a relatively common psychiatric comorbidity of most neurological disorders, with prevalence rates ranging between 20 and 50% among patients with stroke, multiple sclerosis, epilepsy, Parkinson's disease and dementia. Furthermore, depression is an independent predictor of poor quality of life in these patients and has a negative impact on the response to treatment, course and recovery of neurological deficits. Thus, treatment of depression has become an integral part of the management of these neurologic disorders. This article discusses the rationale for neurologists to be trained in recognizing depressive disorders in neurologic patients and identifies the type of mood disorders in which neurologists can provide pharmacotherapy and those that need to be referred to the care of the psychiatrist. PMID- 15820337 TI - Seizure-prone EL/Suz mice exhibit physical and motor delays and heightened locomotor activity in response to novelty during development. AB - Seizure-prone EL/Suz mice have been studied as a model of multifactorial epilepsy for five decades. In prior behavioral studies, EL/Suz mice were shown to exhibit heightened locomotor activity, which implies a state of underlying hyperexcitability. The aim of the present study was to establish the premorbid behavioral development of basic motor skills and activity levels of EL/Suz mice, as compared with DDY mice, the control strain that is not seizure-prone. EL/Suz and DDY pups were monitored from Postnatal Day (PND) 3 to assess body weight, surface righting, negative geotaxis, forelimb grip strength, eye opening, habituation to a novel environment, and exploratory behavior in a two-compartment task. EL/Suz mice weighed less from PNDs 3 to 21 and exhibited delayed surface righting (PNDs 3, 5, 7) and negative geotaxis (PNDs 5, 7, 9) responses. EL/Suz and DDY mice differed in their habituation to a novel environment, with EL/Suz mice exhibiting higher activity, both within a single 10-minute session and across the 3 days of testing. EL/Suz and DDY mice also differed in the two compartment task, with EL/Suz mice exhibiting increased locomotor activity and spending a greater amount of time in the light compartment. Thus, the present findings reveal that EL/Suz mice exhibit some developmental delays, altered habituation to a novel environment, and increased exploratory activity. Overall, the present results demonstrate that the behavioral and physiological phenotype of seizure-prone EL/Suz mice is deviant more than 2 months before the onset of seizure susceptibility. PMID- 15820338 TI - Actions at GABA(A) receptors in the hippocampus may mediate some antiseizure effects of progestins. AB - Progestins can have antiseizure effects; however, the mechanisms and sites of action of these effects are not well-understood. Whether progesterone's actions at GABA(A) receptors in the hippocampus are important for its antiseizure effects was investigated. In Experiment 1, ovariectomized rats were administered sesame oil vehicle or a regimen of progesterone (500 microg sc, which produces physiological concentrations in plasma and the hippocampus), followed 2.5 hours later by administration of saline vehicle or a regimen of bicuculline (1 mg/kg, sc), a GABA(A) receptor antagonist, which does not produce any intrinsic effects on seizures. Progesterone, compared with vehicle, significantly increased the latency to, and decreased the number of, pentylenetetrazole-induced tonic seizures and increased GABA-stimulated chloride flux. Co-administration of bicuculline attenuated progesterone's antiseizure effects and decreased GABA stimulated chloride flux in the hippocampus. Bicuculline did not alter ictal behavior compared with vehicle. In Experiment 2, ovariectomized rats were subcutaneously administered sesame oil or progesterone (500 microg), followed 2.5 hours later by bilateral infusions of bicuculline (100 ng) or vehicle (saline) into the hippocampus. Infusion of bicuculline into the hippocampus of progesterone-primed rats significantly increased ictal activity, compared with that induced by progesterone administration alone, but alone did not alter seizures compared with that produced by saline infusions into the hippocampus. These data suggest that actions of progesterone at GABA(A) receptors in the hippocampus are important for progesterone's antiseizure effects. PMID- 15820339 TI - Correlation between shaking behaviors and seizure severity in five animal models of convulsive seizures. AB - Wet dog shakes (WDS) and head shakes (HS) are associated with experimentally induced convulsive seizures. We sought to determine whether these behaviors are correlated or not with major (status epilepticus (SE) or fully kindled animals) or minor (non-SE or partially kindled animals) seizure severity. WDS are directly correlated with SE induced by intracerebral star fruit extract (Averrhoa carambola) injection and with kindled animals in the amygdala fast kindling model. On the other hand, WDS are inversely correlated with SE induced by intracerebral bicuculline and pilocarpine injections. Systemic pilocarpine in animals pretreated with methyl-scopolamine barely induced WDS or HS. The role of shaking behaviors may vary from ictal to anticonvulsant depending on the experimental seizure model, circuitries involved, and stimulus intensity. The physical presence of acrylic helmets may per se inhibit the HS response. Also, methyl-scopolamine, a drug incapable of crossing the blood-brain barrier, can induce HS in animals without acrylic helmets. PMID- 15820340 TI - Verbal memory outcome in patients with normal preoperative verbal memory and left mesial temporal sclerosis. AB - PURPOSE: Previous studies have shown that structural integrity (i.e., presence/absence of mesial temporal sclerosis (MTS)) of the left mesial temporal lobe is associated with verbal memory outcome following left anterior temporal lobectomy (ATL). However, the functional integrity of the left temporal lobe, as exemplified by preoperative verbal memory performance, has also been associated with verbal memory outcome following surgery. We investigated the risk of verbal memory loss in patients with known structural abnormality (i.e., left mesial temporal sclerosis by MRI) and normal preoperative verbal memory performance who undergo left ATL. METHODS: Seventeen patients with left temporal lobe epilepsy, MRI-based exclusive left MTS, and normal preoperative verbal memory were identified. Normal verbal memory was defined as performance on both Acquisition (learning across trials 1-5) and Retrieval (long delayed free recall) portions of the California Verbal Learning Test (CVLT) above a T score of 40 (>16%ile). Postoperative verbal memory outcome was established by incorporating standardized regression-based (SRB) change scores. RESULTS: Postoperative declines across both CVLT Retrieval T scores and Acquisition T scores (average 20% and average 15% declines from baseline scores, respectively) were measured for the group. The average CVLT Retrieval SRB change score was -2.5, and the average CVLT Acquisition SRB change score was -1.0. A larger proportion of patients demonstrated postoperative declines on Retrieval scores than Acquisition scores (64.7% vs 17.6%, respectively). CONCLUSIONS: Even in the presence of left MTS, patients exhibiting normal presurgical verbal memory are at risk for verbal memory declines following ATL. These results suggest that the functional integrity of the left mesial temporal lobe may play an important role in the verbal memory outcome in this patient group. PMID- 15820341 TI - Prevalence of and risk factors for anxiety and depressive disorders in Nigerian adolescents with epilepsy. AB - The emotional response of adolescents to a chronic illness like epilepsy may differ across cultures. This study was aimed at investigating the prevalence of and risk factors for anxiety and depressive disorders in a group of Nigerian adolescents with epilepsy. Adolescents with epilepsy (n=102) aged between 12 and 18 were assessed for anxiety and depressive disorders with the Diagnostic Interview Schedule for Children Version IV (DISC-IV). An anxiety disorder was diagnosed in 32 (31.37%) of the adolescents and a depressive disorder was reported in 29 (28.43%). Predictors of anxiety and depressive disorders by regression analysis include uncontrolled seizures, polytherapy, and felt stigma. Family factors such as parents' psychopathology and family stress are also moderately significant. Results show that emotional disorders in adolescents with epilepsy cut across cultures. Adequate monitoring, education targeted at reducing felt stigma, and family intervention programs are needed for early intervention. PMID- 15820342 TI - Dimensions of the epilepsy foundation concerns index. AB - We performed principal component analysis (PCA) of the Epilepsy Foundation Concerns Index scale in 189 patients undergoing evaluation for epilepsy surgery. We identified a five-factor solution in which there were no varimax-rotated factors consisting of fewer than two questions. Factor 1 reflects affective impact on enjoyment of life, Factor 2 reflects general autonomy concerns, Factor 3 reflects fear of seizure recurrence, Factor 4 reflects concern of being a burden to one's family, and Factor 5 reflects a perceived lack of understanding by others. Multiple regression using the Quality of Life in Epilepsy--89 question version; Minnesota Multiphasic Personality Inventory--2; Wechsler Adult Intelligence Scale--third edition; and verbal and visual memory tests as predictors demonstrated a different pattern of association with the factor and summary scores. We conclude that the Epilepsy Foundation Concerns Index is multidimensional, and using a global score based on all items may mask specific concerns that may be relevant when applied to individual patients. PMID- 15820343 TI - Temporal lobe ictal behavioral patterns in hippocampal sclerosis and other structural abnormalities. AB - Ictal behavioral characteristics may provide clues in determining the nature of the epileptic focus. We defined ictal behavioral characteristics in patients with intractable temporal lobe epilepsy (TLE) who underwent anterior temporal lobectomy (ATL) and lived seizure-free for 2 years of follow-up. Video/EEG data on 282 seizures observed in 48 patients who suffered from TLE and underwent ATL were analyzed. All patients were seizure-free after surgery. We divided the patients into two groups on the basis of the pathological examination. Two hundred and two seizures in 35 patients with hippocampal sclerosis (Group 1) and eighty seizures in 13 patients with other pathological findings, such as tumors, cavernoma, and hamartoma (Group 2), were analyzed. Ictal behavior characteristics were evaluated for each of the seizures recorded in the two groups. Behavioral arrest, bilateral hand automatisms, oral and leg automatisms, and ictal aggression were significantly more frequent in Group 2 (P<0.05), whereas contralateral dystonia of the upper extremity (P<0.05), ipsilateral hand automatisms (P<0.05), ipsilateral hand automatisms in the presence of contralateral dystonia of the upper extremity (P<0.001), contralateral forced head deviation (P<0.05), and secondary generalization (P<0.05) were more significant in Group 1. There was no significant difference in vocalization and ipsilateral nonforced head deviation between the two groups (P>0.05). The number of seizures observed during ictal speech, crying, and postictal nose wiping was not large enough, so differences could not be analyzed. It was concluded that although ictal behavioral characteristics differed between the two groups, certain behavioral patterns may be helpful in differentiating between hippocampal sclerosis and other pathology. PMID- 15820344 TI - Sexual dysfunction, sex steroid hormone abnormalities, and depression in women with epilepsy treated with antiepileptic drugs. AB - Women with epilepsy are believed to be at risk for sexual dysfunction. Disorders of sexual desire and sexual arousal, including dyspareunia, vaginismus, and lack of lubrication, affect an estimated 30 to 60% of women with epilepsy. In this study, 57 reproductive-aged women with either localization related (LRE) or primary generalized epilepsy (PGE) on antiepileptic drug (AED) monotherapy and 17 nonepileptic controls completed questionnaires examining sexual experience, arousability, anxiety, and symptoms, as well as an inventory of depression. An endocrine assessment was performed during the early follicular phase of the menstrual cycle. Sexual dysfunction was more common in women with LRE, in women receiving phenytoin, in women with low levels of estradiol and dehydroepiandrosterone sulfate, and in women with self-reported symptoms of mild depression. The mechanisms of sexual dysfunction in women with epilepsy are multifactorial, but AED choice appears to be one cause that is modifiable. PMID- 15820345 TI - Ketter's hypothesis of the mood effects of antiepileptic drugs coupled to the mechanism of action of topiramate and levetiracetam. AB - Mood-modulating profiles of antiepileptic drugs (AEDs) have been classified by Ketter, Post, and Theodore [Neurology 1999; 53 (5, Suppl. 2) S53-76] into two classes: the first class is assumed to have deactivating effects related to GABA potentiation, and the second class is assumed to have activating effects that are associated with glutamate attenuation. We tested this hypothesis by reviewing the multiple mechanisms of action of topiramate (TPM) and levetiracetam (LEV) together with clinical behavioral side effects of patients who had been treated with TPM and LEV in a tertiary referral center for epilepsy. We found LEV to manifest activating and deactivating side effects equally and TPM to act as a deactivating AED, with tiredness/sleepiness side effects being predominant. TPM, in comparison to LEV, was found to be associated with a high incidence of side effects. Testing the hypothesis of Ketter et al. (1999) the deactivating effects of TPM may be coupled to a predominance of potentiation of GABA, but the oversimplified basis of the model needs to be acknowledged. PMID- 15820346 TI - Efficacy and cognitive side effects of tiagabine and topiramate in patients with epilepsy. AB - OBJECTIVE: Whereas the efficacy of the newer antiepileptic drugs (AEDs) is well established, there remain questions regarding their cognitive side effects. Therefore, we performed a comparative open randomized trial with TPM and TGB as add-on therapy, with particular consideration of cognition, mood, and health related quality of life (HRQOL). METHODS: Forty-one patients with refractory epilepsy were randomly assigned to one of the two treatment groups (TPM vs TGB) and received neuropsychological testing at baseline (T1), after titration (3 months, T2), and during the maintenance phase (another 3 months, T3). Tests included measures of intelligence, attention, working memory, episodic memory, language, and self-report questionnaires regarding mood and HRQOL. Twenty patients (8 TPM, 12 TGB) discontinued the trial for different reasons (no group difference). RESULTS: Seizure outcome (intention-to-treat analysis) was comparably good in both groups (8.1% seizure free, 29.7% seizure reduction>50%). From baseline to after the titration paired sample t tests revealed significant deterioration in verbal fluency, language comprehension, working memory, and visual block tapping under TPM and a deterioration in verbal memory (delayed free recall) in the TGB group. These functions remained stable in the maintenance phase. Self-report measures initially indicated concerns about AED side effects in both groups and concerns about worse cognitive functioning and depression under TPM. In the maintenance phase the TGB group reported feeling a lack of energy, whereas patients on TPM demonstrated improvement on all QOLIE scales on a descriptive level. CONCLUSION: This study demonstrates the comparable efficacy of TPM and TGB. Consistent with previous reports, TPM but not TGB appears to be associated with persistent negative cognitive side effects on frontal lobe associated functions, the degree of which may be estimated by the fact that this effect was observed with a very small sample size. In contrast, in patients taking TPM, initially negatively affected HRQOL returns to baseline in the long run on a descriptive level. The latter finding may be interpreted in accordance with the observation that objective performance and subjective self-report under TPM can be dissociated. PMID- 15820347 TI - Pharmacological outcomes in newly diagnosed epilepsy. AB - The response to antiepileptic drugs (AEDs) has been examined in 780 adult and adolescent patients with newly diagnosed epilepsy presenting with a range of seizure types and epilepsy syndromes over a 20-year period. Carbamazepine (CBZ, n=312), sodium valproate (VPA, n=315), and lamotrigine (LTG, n=249) were the most common AEDs prescribed as monotherapy. More patients with localization-related epilepsies became seizure free with LTG (63%) than with CBZ (45%, P=0.006) or VPA (42%, P=0.006). For idiopathic generalized epilepsies a greater proportion of patients achieved control with VPA (68%) than with CBZ (31%) or LTG (45%). In particular, more patients with juvenile myoclonic epilepsy responded to VPA (75%) compared with LTG (39%, P=0.014). Seizure freedom was achieved with modest or moderate daily doses (median CBZ 400mg, VPA 1000 mg, (LTG) 150 mg) of all three AEDs in the majority of patients achieving remission. Time to first seizure did not differ among these three drugs when given as first treatment. Adverse effects leading to withdrawal were more frequent with CBZ (16%) than with VPA (7%, P=0.03) or LTG (7%, P=0.018). In patients failing initial monotherapy, response to a combination of two AEDs (27%) was not different from that with alternative monotherapy (32%). The majority of patients with newly diagnosed epilepsy responding to treatment did so rapidly and completely with moderate doses of AEDs, with no differences in time to first seizure. PMID- 15820348 TI - Celiac disease-related antibodies in an epilepsy cohort and matched reference population. AB - Celiac disease (CD) is a common immunologically mediated enteropathic condition manifesting in genetically predisposed individuals after exposure to wheat gluten. CD may be associated with a number of neurological disorders, and the prevalence of epilepsy is reported to be increased in patients with CD. We studied the prevalence of CD-associated antibodies in a cohort of 968 patients with epilepsy and in a population-based reference group of 584 subjects. There was no difference in the prevalence of IgA and IgG class antigliadin antibodies (AGAbA and AGAbG, respectively), endomysial antibodies (EMA), and tissue transglutaminase antibodies (tTGAbA) between patients with epilepsy and the control population. This suggests the presence of CD to be the same in patients with epilepsy and the reference population. However, AGAbA was more prevalent in patients with primary generalized epilepsy (19.6%) than in the reference population (10.6%, relative risk 1.8, 95% confidence interval 1.1-3.0, P=0.012). PMID- 15820349 TI - Cognitive functioning in bilateral perisylvian polymicrogyria (BPP): clinical and radiological correlations. AB - Bilateral perisylvian polymicrogyria (BPP) is a malformation of cortical development, frequently associated with severe dysarthria or anarthria. BPP patients are therefore often labeled as severely retarded, but a detailed neuropsychological profile has not been reported to date. In a series of 14 patients, we demonstrated that only a minority had extremely low intelligence, and that some aspects of cognitive function correlated with the extent of the cortical disorganization. Early age at seizure onset correlated positively with Performance IQ scores (P<0.05) and negatively with the extent of the lesion (P<0.01), reflecting that patients with more severe BPP are more likely to have early seizure onset, resulting in greater interference with ongoing cognitive development. Receptive and expressive language skills were found to be equally poor. Frontal lobe function and memory abilities were relatively well preserved, suggesting that the observed cognitive profiles were related, at least in part, to specific areas of cortical dysfunction and not only to global dysfunction. PMID- 15820350 TI - A pilot study of the effect of Direct Instruction programming on the academic performance of students with intractable epilepsy. AB - Fifty-five students with intractable epilepsy and learning difficulties, aged 6.5 to 14.1 years with a mean IQ of 71.25 (SD=13.56), attended a hospital-based classroom of no more than eight students where they received Direct Instruction programming for 3 to 4.5 days a week, typically for up to 16 weeks. Pretest posttest academic gains were evaluated in comparison to normative test performance. At pretest the students were achieving below test means in reading and mathematics, particularly in calculation. Direct Instruction was associated with significant improvement in all academic areas except word identification in reading. IQ level was related to certain academic gains, but gains were not associated with seizure variables or the number of days students spent in the program. Despite severe learning difficulties, students with intractable epilepsy can make academic gains when taught by highly structured teaching methods such as Direct Instruction. PMID- 15820351 TI - Primidone is associated with interictal depression in patients with epilepsy. AB - Depressive symptoms are common in epilepsy. To determine associations between depression and demographic, clinical, and pharmacological factors among epileptic patients, we conducted a cross-sectional survey. We evaluated 241 epileptic outpatients at a neurological center in a 6-month period. Depressive syndrome was diagnosed when both the Montgomery-Asberg Scale and the Beck Depression Inventory were rated above the standard cutoff points. Bivariate and multivariate analyses were performed to assess the differences between depressed and nondepressed patients with respect to demographic, clinical, and pharmacological features. Depressive syndrome was diagnosed in 42.7% of patients (n=103). Factors associated in the bivariate analysis were: cryptogenic etiology, posttraumatic epilepsy, use of primidone, and inadequate seizure control. After logistic regression, inadequate seizure control (OR 3.08, 95% CI 1.40-6.77, P=0.005) and use of primidone (OR 4.08, 95% CI 2.09-7.98; P<0.001) remained significantly associated. Depression was common and associated with inadequate seizure control and use of primidone. PMID- 15820352 TI - Vagus nerve stimulation therapy: 2-year prospective open-label study of 40 subjects with refractory epilepsy and low IQ who are living in long-term care facilities. AB - Treating seizures among patients with mental retardation/developmental disabilities (MR/DD) is difficult owing in large part to the presence of additional comorbidities and the resulting need for polytherapy. Therefore, a nonpharmacological treatment option is needed for this population. This prospective, open-label study documented the long-term outcome of 40 low-IQ (<70) patients living in long-term care facilities who received vagus nerve stimulation (VNS) therapy for pharmacoresistant epilepsy. Subjects were seen every 1 to 3 months by their neurologist (R.H.). Seizure frequency, antiepileptic medication, and quality-of-life information were documented preimplantation and quarterly thereafter through 2 years. The surgery and therapy were well tolerated. Seizures were reduced by at least 50% for 11 subjects. Antiepileptic medications were reduced from 3.3 per subject at baseline to an average of 2.3 per subject after 2 years. According to caregiver reports, overall quality of life improved for the majority of subjects; also, using the Client Development Evaluation Report (CDER), statistically significant improvements were reported at both 1 and 2 years in attention span, word usage, clarity of speech, standing balance, washing dishes, and household chores. VNS is a viable treatment option for low-IQ patients with pharmacoresistant epilepsy who are living in long-term care facilities. PMID- 15820353 TI - Gender differences in risk factors of suicidal behavior in epilepsy. AB - The current study was performed to determine possible gender differences among risk factors for suicidal behavior in epilepsy. A special rating scale for assessment of suicidality was designed. The risk of suicide attempt was higher in epilepsy with concomitant diagnoses of organic affective disorder (F=06.3, ICD 10) and cognitive impairment (F=07.62, ICD-10). Risk was higher among females than males. Early age of epilepsy onset and high frequency of secondary generalized, simple partial, and all seizures were risk factors for suicidality in males, whereas low frequency of complex partial seizures was the risk factor in females. Daily dose of a classic antiepileptic drug was a risk factor for males, whereas daily dose of phenobarbital was a risk factor for both genders. Daily dose of carbamazepine and valproate is inversely correlated with suicidal behaviour in women, but not in men. The results obtained are discussed in the context of a paradigm of gender dimorphism as an indicator of evolution of Homo sapiens. PMID- 15820354 TI - Do antiepileptic drugs accelerate forgetting? AB - The majority of patients with epilepsy become seizure-free with antiepileptic drug therapy. However, seizures in approximately one-third of patients with epilepsy are difficult to treat with antiepileptic drugs and require high doses or polytherapy. High dosages increase the risk of cognitive side effects. We retrospectively investigated 162 patients with refractory temporal lobe epilepsy to determine whether the antiepileptic drugs carbamazepine, phenobarbital, and phenytoin affect the acquisition and retention of verbal and visual information. We found that patients with high serum levels of these antiepileptic drugs were selectively impaired in the retention but not acquisition of new information. Intelligence, age, duration of epilepsy, and seizure frequency were controlled for and were not factors in the observed results. There were no differences in favor of a certain drug with respect to memory functioning. Our results suggest that patients with refractory epilepsy with high serum levels of the antiepileptic drugs studied are at higher risk of accelerated forgetting. PMID- 15820355 TI - The diagnostic value of EEGs in patients with syncope. AB - We retrospectively reviewed reports of all EEGs performed at the New Jersey Neuroscience Institute at JFK Hospital between January 1999 and December 2003. Of 9234 EEGs performed, 1094 were of patients with syncope. Among patients with syncope, 67.18% of the EEGs were normal and 28.15% showed diffuse and focal slowing. Only 1.46% of the EEGs showed epileptiform discharges (EDs). This is similar to the incidence of EDs in healthy adults. The presence of EDs did not change the management of these patients. Therefore, EEGs have very low yield and should not be routinely obtained in patients with syncope. PMID- 15820356 TI - Childhood paroxysmal kinesigenic dyskinesia: report of seven cases with onset at an early age. AB - We report on seven children who developed abnormal involuntary movements as early as 1.5 years after unremarkable term births. The paroxysmal episodes of abnormal movements were typically precipitated by sudden, voluntary movements, or a startle. The clinical features in each case were consistent with the diagnosis of paroxysmal kinesigenic dyskinesia (PKD). The episodes of abnormal movements are described. EEG was obtained in all cases, and video/electroencephalography (VEEG) monitoring was performed to exclude the possibility of epilepsy in six patients. VEEG studies revealed multiple events consistent with PKD; no ictal epileptiform discharges were recorded. The apparent benign nature of the disorder, as well as treatment options with antiepileptic drugs, was discussed with the parents, and most chose no pharmacologic treatment. We discuss clinical characteristics of PKD, treatment with anticonvulsant therapy, and recent insights into its possible pathophysiology. PMID- 15820357 TI - Presurgical evaluation and cognitive functional reorganization in Fishman syndrome. AB - Fishman syndrome, also known as encephalocraniocutaneous lipomatosis (ECCL), is a rare, congenital neurocutaneous syndrome characterized by unilateral skin, eye, and brain abnormalities. Epileptic seizures and developmental delay are usually present. We report the clinical, radiological, and, for the first time, neurophysiological findings in a 24-year-old woman diagnosed with ECCL who was evaluated for epilepsy surgery. Functional magnetic resonance imaging revealed transfer of memory and language functions to the nonaffected hemisphere, providing evidence that functional reorganization and restoration of cognitive functions may occur in the context of extensive malformations, such as neurocutaneous syndromes. PMID- 15820358 TI - Recruitment rates and fear of phlebotomy in pediatric patients in a genetic study of epilepsy. AB - This study examined participation rates and reasons for refusal in a genetic study of human epilepsy. The study enrolled children with epilepsy and their parents, and required signing informed consent, verbalizing assent, and giving a peripheral blood sample. One hundred sixty-eight children met inclusion criteria; 137 agreed to enroll (82%), and 31 refused (18%). Sixteen of thirty-one patients (52%) who refused cited fear of phlebotomy as the reason for refusal. All patients refusing due to fear of phlebotomy did not require blood tests for clinical purposes. As fear of phlebotomy is the primary reason for study refusal, obtaining DNA samples from a buccal swab or mouthwash protocol may be an alternative for some studies, although there are limitations to these methods. Further analysis of the factors influencing decisions to decline study enrollment is warranted. These data will help in the design of future genetic studies and may increase future participation rates. PMID- 15820359 TI - Pattern-induced partial seizures with repetitive affectionate kissing: an unusual manifestation of right temporal lobe epilepsy. AB - We report a case of recurrent partial seizures that were often precipitated by looking up a flight of stairs and included spitting as well as repetitive affectionate kissing automatisms. These seizures were shown by long-term video/EEG monitoring to be of right temporal origin and completely subsided after right temporal lobectomy. This case is unique because: (1) The patient had partial rather than primarily generalized pattern-induced seizures. (2) Affectionate kissing automatisms were a part of his partial seizures and, to our knowledge, have not been reported in the literature before. PMID- 15820360 TI - Hush sign: a new clinical sign in temporal lobe epilepsy. AB - Neurologists have been analyzing the clinical behaviors that occur during seizures for many years. Several ictal behaviors have been defined in temporal lobe epilepsy (TLE). Ictal behaviors are especially important in the evaluation of epilepsy surgery candidates. We propose a new lateralizing sign in TLE originating from the nondominant hemisphere-the "hush" sign. Our patients were 30 and 21-year old women (Cases 1 and 2, respectively). Their epileptogenic foci were localized to the right mesial temporal region after noninvasive presurgical investigations. Case 1 had no cranial MRI abnormality, whereas cranial MRI revealed right hippocampal atrophy in Case 2. These women repeatedly moved their right index fingers to their mouth while puckering their lips during complex partial seizures. We have named this ictal behavior the "hush" sign. Anterior temporal lobectomy with amygdalohippocampectomy was performed in both patients, and pathological examinations revealed hippocampal sclerosis. The "hush" sign no longer occurred after seizures were controlled. They were seizure free as of 30 and 31 months of follow-up, respectively. We believe that the "hush" sign may be supportive of a diagnosis of TLE originating from the nondominant hemisphere. This sign may occur as a result of ictal activation of a specific brain region in this hemisphere. PMID- 15820361 TI - Prolonged postictal psychosis with forced normalization (Landolt) in temporal lobe epilepsy. AB - We report two patients with temporal lobe epilepsy who developed prolonged episodes of postictal psychosis (12 weeks) following a bout of seizures and a lucid interval. No drug toxicity was noted, and no recurrent or continuous seizures were observed during their prolonged psychotic episodes. Repeated EEGs during the period of psychosis showed that their habitual focal epileptiform abnormalities had disappeared. Responses to neuroleptic treatments were not remarkable. Their psychotic symptoms gradually dispelled after their epileptiform abnormalities reappeared. These observations may suggest possible overlapping mechanisms between a prolongation of psychosis and suppression of epileptiform discharges on a scalp EEG. PMID- 15820362 TI - Perioral myoclonia with absences: an overlooked and misdiagnosed generalized seizure type. AB - OBJECTIVE: The goal of this report is to draw attention to perioral myoclonia with absences, a rare and underrecognized generalized seizure type. METHODS: We describe a 31-year-old, mildly retarded patient who had absence seizures with prominent perioral movements and rare generalized tonic-clonic convulsions. He had an attack of status epilepticus consisting of asymmetrical perioral myoclonic jerks and was misdiagnosed with epilepsia partialis continua. RESULTS: The perioral myoclonia were associated with brief lapses of consciousness and with a generalized EEG pattern as shown by the synchronous video/EEG investigation with polygraphic recordings. The patient was diagnosed as having the syndrome of perioral myoclonia with absences described by C.P. Panayiotopoulos, C.D. Ferrie, S.E. Giannakodimos, and R.O. Robinson (in: Wolf P, editor. Epileptic seizures and syndromes. London: John Libbey; 1994, p. 143-53). CONCLUSION: We emphasize that this rare and peculiar seizure type associated with generalized epilepsy can be misdiagnosed as focal epilepsy. PMID- 15820363 TI - Writing epilepsy: a neurophysiological, neuropsychological and neuroimaging study. AB - Writing epilepsy is a rare reflex syndrome in which seizures are triggered by writing. We describe a 33-year-old, right-handed man, with a history of juvenile absence epilepsy in remission and a family history of epilepsy, in whom myoclonic jerks precipitated exclusively by writing started at the age of 30. Intensive video/EEG monitoring during neuropsychological tests revealed, at about 1 minute after starting to write, a dystonic posture, followed by myoclonic jerks involving the right hand that shortly after became generalized. Concomitantly, the ictal EEG documented generalized hypersynchronous polyspike-wave discharges, maximal over the right parietocentral area. SPECT revealed an ictal hyperperfusion and interictal hypoperfusion over right parietofrontal regions, and fMRI showed extensive and intense left frontal, supplementary motor area activation, induced by writing. This case study provides some evidence supporting the hypothesis that the mechanism underlying writing-triggered seizures may be a generalized seizure process, with a focal cortical trigger zone, presumed to be the left frontal lobe as suggested by clinical and fMRI data. A relevant role played by the right hemisphere (right parietofrontal region) is postulated in the full-blown expression of reflex epileptogenesis, as supported by EEG and SPECT findings. PMID- 15820364 TI - New evidence on birth spacing: promising findings for improving newborn, infant, child, and maternal health. AB - This editorial summarizes new evidence, some of which is published in this supplement, on birth spacing and newborn, infant, child and maternal health, as well as the demand for birth spacing services in the developing world. The article points to the high number of annual infant, child and maternal deaths, low birth weight infants and malnourished infants and children in developing countries. It highlights several new findings on birth spacing relevant to these conditions: It argues that, in light of the new evidence, birth spacing is an important, feasible and practical intervention to address these conditions and should be included in developing country health programs. PMID- 15820365 TI - Effect of interpregnancy interval on birth outcomes: findings from three recent US studies. AB - The relationship between interpregnancy interval and adverse birth outcomes (i.e., low birth weight, preterm birth, and small size for gestational age) was examined in three recent studies conducted in Utah and Michigan of the United States. These studies were conducted among different populations, used different study designs (i.e., cross-sectional and retrospective cohort designs), and addressed several other methodological limitations in the previously published literature. In addition, the data were stratified by, and controlled for, several maternal reproductive risk factors. A J-shaped relationship between interpregnancy interval and adverse birth outcomes was observed in all three studies. The risk for adverse birth outcomes is lowest when the interpregnancy interval was 18-23 months and increased when the interval departed from 18-23 months. This J-shaped relationship existed at levels of maternal reproductive risk factors and after these risk factors were controlled for using logistic regression. Based on the consistency of the findings from all three studies, it appears that the J-shaped relationship between interpregnancy interval and adverse birth outcomes is causal. This information can be used by health care providers and public health programs to counsel and educate women who recently gave births on reducing the risk for adverse birth outcomes by means of appropriate pregnancy spacing. PMID- 15820366 TI - Effect of the interpregnancy interval after an abortion on maternal and perinatal health in Latin America. AB - OBJECTIVE: To investigate whether the length of the interval between an abortion and the next pregnancy is associated with increased risks of adverse maternal and perinatal outcomes in Latin America. METHOD: Retrospective cross-sectional study using information from 258,108 women delivering singleton infants and whose previous pregnancy resulted in abortion recorded in the Perinatal Information System database of the Latin American Centre for Perinatology and Human Development, Montevideo, Uruguay, between 1985 and 2002. Adjusted odds ratios were obtained through logistic regression analysis. RESULT: Compared with the post-abortion interpregnancy intervals of 18 to 23 months, intervals shorter than 6 months were significantly associated with increased risks of maternal anemia, premature rupture of membranes, low birth weight, very low birth weight, preterm delivery, and very preterm delivery. CONCLUSION: In Latin America, post-abortion interpregnancy intervals shorter than 6 months are independently associated with increased risks of adverse maternal and perinatal outcomes in the next pregnancy. DEFINITION: Post-abortion interpregnancy interval (PAII): the time elapsed between the day of the abortion and the first day of the last menstrual period for the index pregnancy. PMID- 15820367 TI - Pregnancy spacing and maternal morbidity in Matlab, Bangladesh. AB - OBJECTIVE: The study examines the relationship between pregnancy spacing and seven measures of maternal morbidity in Matlab, Bangladesh. METHOD: The study analyzes maternal morbidity data on 11,122 women who visited a health center during their third trimester of pregnancy between 1996 and 2002. Adjusted odds ratios were obtained through logistic regression analysis to assess the effects of pregnancy intervals of differing lengths while holding constant other influences (six variables) on maternal morbidity. RESULTS: After controlling these variables, pre-eclampsia and high blood pressure are significantly more likely for women with preceding inter-pregnancy intervals of less than 6 months or 75 months or more compared to those with intervals of 27-50 months. Premature rupture of membranes is significantly more likely following inter-pregnancy intervals of 6-14 months, and edema is significantly more likely following inter pregnancy intervals over 50 months. CONCLUSION: Short and long inter-pregnancy intervals are associated with increased incidence of some maternal morbidities. PMID- 15820368 TI - Existing demand for birth spacing in developing countries: perspectives from household survey data. AB - OBJECTIVE: To identify the extent of demand for birth spacing, according to age and parity among married women of reproductive age (MWRA) in developing countries. METHODS: Secondary analysis of data from the Demographic and Health Surveys (DHS) using cross-tabulations. Data collected from nationally representative samples of MWRA in selected developing countries between 1990 and 2004. RESULTS: Demand for birth spacing is the most prevalent reason for an interest in family planning among married women aged 15-29 years in the majority of developing countries examined. In the 15-19-year age cohort, the demand for spacing is proportionally the most prevalent reason for a demand for family planning. A demand for spacing even exists among young, zero-parity married women in each country examined. Findings on the demand for spacing among zero-parity married women quantifies the expressed desire of some married women in developing countries to postpone a first birth or the timing of a first pregnancy. CONCLUSION: The substantial demand for birth spacing among young, low- and zero parity women suggests that family planning programs in developing countries may need to reevaluate how accessible services are for this cohort of potential contraception users. Currently, many service-delivery protocols, counseling practices and service provider training may not fully address the needs of younger, low- or zero-parity clients. PMID- 15820369 TI - Effects of preceding birth intervals on neonatal, infant and under-five years mortality and nutritional status in developing countries: evidence from the demographic and health surveys. AB - OBJECTIVE: This paper examines the association between birth intervals and infant and child mortality and nutritional status. METHODS: Repeated analysis of retrospective survey data from the Demographic and Health Surveys (DHS) program from 17 developing countries collected between 1990 and 1997 were used to examine these relationships. The key independent variable is the length of the preceding birth interval measured as the number of months between the birth of the child under study (index child) and the immediately preceding birth to the mother, if any. Both bivariate and multivariate designs were employed. Several child and mother-specific variables were used in the multivariate analyses in order to control for potential bias from confounding factors. Adjusted odds ratios were calculated to estimate relative risk. RESULTS: For neonatal mortality and infant mortality, the risk of dying decreases with increasing birth interval lengths up to 36 months, at which point the risk plateaus. For child mortality, the analysis indicates that the longer the birth interval, the lower the risk, even for intervals of 48 months or more. The relationship between chronic malnutrition and birth spacing is statistically significant in 6 of the 14 surveys with anthropometric data and between general malnutrition and birth spacing in 5 surveys. However, there is a clear pattern of increasing chronic and general undernutrition as the birth interval is shorter, as indicated by the averages of the adjusted odds ratios for all 14 countries. CONCLUSION: Considering both the increased risk of mortality and undernutrition for a birth earlier than 36 months and the great number of births that occur with such short intervals, the author recommends that mothers space births at least 36 months. However, the tendency for increased risk of neonatal mortality for births with intervals of 60 or more months leads the author to conclude that the optimal birth interval is between 36 and 59 months. This information can be used by health care providers to counsel women on the benefits of birth spacing. PMID- 15820370 TI - Pediatric rheumatology. PMID- 15820371 TI - Inflammation in juvenile idiopathic arthritis. AB - Inflammation evolved to aid in the clearance of microorganisms. In pediatric arthritides, the inflammation persists and causes damage to the joint. The contribution of the innate immune system to inflammation is significant and can be exploited therapeutically. Although cells of the adaptive immune system such as T cells and B cells participate in the disease process, many of the features of arthritis are directly attributable to inflammatory mediators. Recent advances in the understanding of these processes have led to dramatic improvements in treatment. PMID- 15820372 TI - Measurement of health status, functional status, and quality of life in children with juvenile idiopathic arthritis: clinical science for the pediatrician. AB - Several groups have undertaken research on health status, functional status, and quality of life in the pediatric rheumatic diseases, particularly juvenile idiopathic arthritis (JIA) and juvenile rheumatoid arthritis. This article highlights the principles involved in this type of measurement, discusses the measures that have been developed for JIA, and describes the outcomes determined from recent retrospective and prospective longitudinal outcome studies. These studies suggest that although there has been improvement in overall outcomes, significant numbers of individuals persist with active disease into adulthood and have significant damage, reduced functional ability, and disability. PMID- 15820373 TI - Radiologic investigation of rheumatic diseases. AB - This article reviews the current use of the wide variety of imaging modalities now available, presenting the imaging features of common and important causes of acute and chronic rheumatic disorders including juvenile idiopathic arthritis, spondyloarthropathies/enthesitis-related arthritis, sepsis, autoimmune diseases, vasculitis, and osteoporosis. PMID- 15820374 TI - Juvenile idiopathic arthritis. AB - Juvenile idiopathic arthritis (JIA), a term referring to a group of disorders characterized by chronic arthritis, is the most common chronic rheumatic illness in children and is a significant cause of short- and long-term disability. This article discusses the classification, differential diagnosis, and treatment of JIA. PMID- 15820375 TI - Systemic lupus erythematosus. AB - Systemic lupus erythematosus in children and adolescents is a multisystem autoimmune disease with a great variability in disease presentation and course. This article summarizes available epidemiologic data, clinical patterns, approaches to investigation and treatment, and recent outcome data. PMID- 15820376 TI - Antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is a systemic autoimmune disorder characterized by a combination of arterial or venous thrombosis and recurrent fetal loss, accompanied by elevated titers of antiphospholipid antibodies (aPL). APS has been recognized as the leading cause of vascular thrombosis in children. The diagnostic and therapeutic approach to APS in childhood may be different from that for adults and because of the rarity of aPL-related thrombosis in children, the natural history and optimal management of APS can be defined only through large, multicenter, controlled studies. PMID- 15820377 TI - Inflammatory myopathies in children. AB - Juvenile idiopathic inflammatory myopathies are rare conditions that are probably autoimmune in nature. Juvenile dermatomyositis (JDM) is the most common inflammatory myopathy. This article describes a recent patient who presented with typical JDM and uses her case to discuss aspects of the childhood inflammatory myopathies. PMID- 15820378 TI - Scleroderma in children. AB - This article discusses the various clinical, laboratory, and therapeutic aspects of systemic sclerosis and localized scleroderma in children. The close collaboration among pediatricians, rheumatologists, and dermatologists represents an important advance in the management of these disabling conditions. PMID- 15820379 TI - Vasculitis in children. AB - This article provides a general overview of vasculitis, situations in which the diagnosis should be considered, diagnostic methods, and therapeutic considerations. Details and treatments unique to specific vasculitides are also reviewed. PMID- 15820380 TI - Periodic fever syndromes. AB - Human autoinflammatory diseases (except for the periodic fever, adenopathy, pharyngitis, aphthae syndrom) are a heterogeneous group of genetically determined diseases characterized by seemingly unprovoked inflammation, in the absence of autoimmune or infective causes. Tremendous advances in the understanding of these disorders have been seen in the last decade. This article discusses hereditary autoinflammatory syndromes that are associated with recurrent fevers. PMID- 15820381 TI - Pediatric pain syndromes and management of pain in children and adolescents with rheumatic disease. AB - This article introduces important issues related to pain in children with musculoskeletal pain syndromes and rheumatic disease, using juvenile primary fibromyalgia syndrome (JPFS) and juvenile idiopathic arthritis (JIA) as models. A brief summary of the prevalence of pain in healthy children is followed by a summary of existing pain-assessment techniques. The remainder of the article describes the pain experience of children with JPFS and JIA and discusses issues related to pain management. PMID- 15820382 TI - Transition of the adolescent patient with rheumatic disease: issues to consider. AB - This article discusses the definitions of transition and models for providing transition care to youth and young adults with rheumatic diseases in the context of differing cultural and health care systems. Issues and questions relating to pediatric rheumatology transition programming are outlined, and a model program of pediatric rheumatology transition is presented. PMID- 15820383 TI - Calcium in the function of the nervous system: new implications. PMID- 15820384 TI - Calcium transients and neural induction in vertebrates. AB - Evidence indicates that a variety of different types of Ca2+ transients (i.e., standing gradients, pulses and propagating waves) may be occurring both simultaneously and sequentially during neural induction in vertebrate embryos. Transients have been observed in the dorsal marginal zone and in the presumptive neural ectoderm, suggesting that they may be generated within two distinct germ layers at separate embryological locations. It has been proposed that the Ca2+ transients might have multiple roles during the period of neural induction, ranging from: activating the expression of early neural genes; contributing to the inhibition of BMP-4 signalling; generating secretion gradients of morphogens; regulating and co-ordinating convergent extension; and establishing and reinforcing dorsoventral axis specification. Both intra- and extracellular stores (either acting separately or in combination) have been shown to generate the neuralizing Ca2+ transients via well-established release mechanisms, and transients have been shown to propagate between connected cells, suggesting an intercellular signalling dimension. Thus, good evidence is accumulating to suggest that Ca2+ might be a key central regulator in the process of neural induction. PMID- 15820385 TI - Ca2+ transients control CNS neuronal migration. AB - In the developing CNS, postmitotic neurons exhibit dynamic changes in the mode, direction and rate of migration as they traverse different cortical layers, but the mechanisms underlying this process is largely unknown. Recent studies show that the changes in Ca2+ transient frequency play a central role in controlling the neuronal cell migration in a cortical layer-specific manner. In this article, we will first describe how granule cells migrate through different terrains of the developing cerebellar cortex. We will then present how such migration of granule cells is controlled by altering the Ca2+ transient frequency in their somata. Finally, we will discuss how the loss of Ca2+ transients triggers the completion of granule cell migration at their final destination. PMID- 15820386 TI - Calcium signalling in growth cone migration. AB - Growth cones, the motile structures at the tips of advancing axons and dendrites, respond to a wide range of cues by either turning towards or away from the cue. Cytosolic calcium signals appear to mediate a large fraction of both types of response. Calcium signals can be generated by influx through plasma membrane channels or by release from intracellular stores. While neurotransmitters can elicit calcium influx through ionotropic receptors, other chemical cues open plasma membrane voltage gated calcium channels by a mechanism other than a change of membrane voltage. In general attractive cues generate spatially and temporally restricted calcium increases that are difficult to detect using conventional indicators. One target for these calcium signals is calmodulin dependent protein kinase II. Repulsive cues generate spatially and temporally more diffuse calcium increases that can be more readily detected using fluorescent indicators. One target for these is the phosphatase calcineurin, which may act by dephosphorylating GAP43 and allowing the latter to cap actin filaments. PMID- 15820387 TI - Regulation of dendritic growth and plasticity by local and global calcium dynamics. AB - The dendritic arbors of neurons are organized into unique spatial patterns that are well suited for their specific functions. Although the intracellular signaling pathways that determine dendritic arbor size and branching patterns during development are not completely understood, it is evident that neurotransmission-mediated elevation in intracellular calcium levels ([Ca2+]i) plays a major role. Recent advances in calcium imaging and genetic approaches have provided new insight into how calcium acts to regulate dendritic growth and remodeling. Global increases in [Ca2+]i that occur upon neuronal depolarization control dendritic arbor growth by regulating transcription. However, the detailed branching patterns of dendritic arbors are regulated by local calcium signaling. Calcium-induced calcium release (CICR) from intracellular stores acts to locally stabilize dendritic branches, presumably triggered by neurotransmission upon contact with newly established inputs. The effects of global and local increases in [Ca2+]i on dendritic structure are cell type- and age-dependent. Unraveling the signaling pathways evoked by local and global rises in [Ca2+]i that shape the developing dendritic arbor at each developmental stage remains challenging but highly important. PMID- 15820389 TI - Homeostatic activity-dependent paradigm for neurotransmitter specification. AB - Calcium-signaling plays a central role in specification of the chemical transmitters neurons express, adjusting the numbers of cells that express excitatory and inhibitory transmitters as if to achieve homeostatic regulation of excitability. Here we review the extent to which this activity-dependent regulation is observed for a range of different transmitters. Strikingly the homeostatic paradigm is observed both for classical and for peptide transmitters and in mature as well as in embryonic nervous systems. Transmitter homeostasis adds another dimension to homeostatic regulation of function in the nervous system that includes regulation of levels of voltage-gated ion channels, densities of neurotransmitter receptors, and synapse numbers and strength. PMID- 15820388 TI - Regulation of dendritic development by calcium signaling. AB - Neuronal activity can have profound effects on dendrite morphology in the developing brain. The effects of neuronal activity on dendritic morphology are mediated by calcium signaling. While many effects of calcium on dendrite structure occur locally at the site of calcium entry into the cytoplasmic milieu, elevation of cytoplasmic calcium is also translated into changes in gene transcription. Decoding the calcium signal into specific changes in gene transcription involve coordinating the action of a number of kinases, phosphatases, transcription factors and transcriptional coactivators. This review focuses on the contribution of calcium-dependent transcription on the control of dendritic morphology. PMID- 15820390 TI - Retinal waves: mechanisms and function in visual system development. AB - A characteristic feature of developing neural networks is spontaneous periodic activity. In the developing retina, retinal ganglion cells fire bursts of action potentials that drive large increases in intracellular calcium concentration with a periodicity of minutes. These periodic bursts of action potentials propagate across the developing inner retina as waves, driving neighboring retinal ganglion cells to fire in a correlated fashion. Here we will review recent progress in elucidating the mechanisms in mammals underlying retinal wave propagation and those regulating the periodicity with which these retinal waves occur. In addition, we will review recent experiments indicating that retinal waves are critical for refining retinal projections to their primary targets in the central visual system and may be involved in driving developmental processes within the retina itself. PMID- 15820391 TI - Temporal integration of intracellular Ca2+ signaling networks in regulating gene expression by action potentials. AB - Temporal aspects of intracellular calcium signaling are particularly important in activity-dependent regulation of gene expression in neurons. This review traces calcium-dependent intracellular signaling from the membrane to the nucleus in response to action-potential firing, and considers how specific genes are regulated by specific patterns of impulse firing. Modes of calcium influx, calcium-dependent protein kinases, transcription factors, individual genes, and genomic analysis are examined, with particular emphasis on the importance of temporal aspects of calcium dynamics in regulating these processes. PMID- 15820392 TI - Calcium imaging of network function in the developing spinal cord. AB - We have used calcium imaging to visualize the spatiotemporal organization of activity generated by in vitro spinal cord preparations of the developing chick embryo and the neonatal mouse. During each episode of spontaneous activity, we found that chick spinal neurons were activated rhythmically and synchronously throughout the transverse extent of the spinal cord. At the onset of a spontaneous episode, optical activity originated in the ventrolateral part of the cord. Back-labeling of spinal interneurons with calcium dyes suggested that this ventrolateral initiation was mediated by activation of a class of interneurons, located dorsomedial to the motor nucleus, that receive direct monosynaptic input from motoneurons. Studies of locomotor-like activity in the anterior lumbar segments of the neonatal mouse cord revealed the existence of a rostrocaudal wave in the oscillatory component of each cycle of rhythmic motoneuron activity. This finding raises the possibility that the activation of mammalian motoneurons during locomotion may share some of the same rostrocaudally organized mechanisms that evolved to control swimming in fishes. PMID- 15820393 TI - Calcium imaging of cortical networks dynamics. AB - Studies relating spontaneous network activities to cognitive processes and/or brain disorders constitute a recently expanding field of investigation. They are mostly based either on cellular recordings--usually performed in pharmacologically induced oscillations in brain slices--or on multi-cellular recordings using tetrodes or multiple electrodes. However, these research strategies cannot link the electrical recordings with morphological characterization of the neurons. The progress made in imaging techniques allows for the first time to have simultaneously a dynamic and global characterization of network activity and to determine the single-cell properties of the unitary microcircuits involved in this activity. PMID- 15820394 TI - Determinants of postsynaptic Ca2+ signaling in Purkinje neurons. AB - Neuronal integration in Purkinje neurons involves many forms of Ca2+ signaling. Two afferent synaptic inputs, the parallel and the climbing fibers, provide a major drive for these signals. These two excitatory synaptic inputs are both glutamatergic. Postsynaptically they activate alpha-amino-3-hydroxy-5-methyl-4 propionic acid (AMPA) receptors (AMPARs) and metabotropic glutamate receptors (mGluRs). Unlike most other types of central neurons, Purkinje neurons do not express NMDA (N-methyl-D-aspartate) receptors (NMDARs). AMPARs in Purkinje neurons are characterized by a low permeability for Ca2+ ions. AMPAR-mediated synaptic depolarization may activate voltage-gated Ca2+ channels, mostly of the P/Q-type. The resulting intracellular Ca2+ signals are shaped by the Ca2+ buffers calbindin and parvalbumin. Ca2+ clearance from the cytosol is brought about by Ca2+-ATPases in the plasma membrane and the endoplasmic reticulum, as well as the Na+-Ca2+-exchanger. Binding of glutamate to mGluRs induces postsynaptic Ca2+ transients through two G protein-dependent pathways: involving (1) the release of Ca2+ ions from intracellular Ca2+ stores and (2) the opening of the cation channel TRPC1. Homer proteins appear to play an important role in postsynaptic Ca2+ signaling by providing a direct link between the plasma membrane-resident elements (mGluRs and TRPC1) and their intracellular partners, including the IP3Rs. PMID- 15820395 TI - Calcium dynamics in dendritic spines, modeling and experiments. AB - Dendritic spines are microstructures, about one femtoliter in volume, where excitatory synapses are made with incoming afferents, in most neurons of the vertebrate brain. The spine contains all the molecular constituents of the postsynaptic side of the synapse, as well as a contractile element that can cause its movement in space. It also contains calcium handling machineries to allow fast buffering of excess calcium that influx through voltage and NMDA gated channels. The spine is connected to the dendrite through a thin neck that serves as a variable barrier between the spine head and the parent dendrite. We review a novel modeling approach that is more suitable for the accurate description of the stochastic behavior of individual molecules in microstructures. Using this approach, we predict the calcium handling ability of the spine in complex situations associated with synaptic activity, spine motility and plasticity. PMID- 15820396 TI - Calcium regulation of actin dynamics in dendritic spines. AB - Most excitatory synapses in the brain are made on spines, small protrusions from dendrites that exist in many different shapes and sizes. Spines are highly motile, a process that reflects rapid rearrangements of the actin cytoskeleton inside the spine, and can also change shape and size over longer timescales. These different forms of morphological plasticity are regulated in an activity dependent way, involving calcium influx through glutamate receptors and voltage gated calcium channels. Many proteins regulating the turnover of filamentous actin (F-actin) are calcium-dependent and might transduce intracellular calcium levels into spine shape changes. On the other hand, the morphology of a spine might affect the function of the synapse residing on it. In particular, the induction of synaptic plasticity is known to require large elevations in the postsynaptic calcium concentration, which depend on the ability of the spine to compartmentalize calcium. Since the actin cytoskeleton is also known to anchor postsynaptic glutamate receptors, changes in the actin polymerization state have the potential to influence synaptic function in a number of ways. Here we review the most prominent types of changes in spine morphology in hippocampal pyramidal cells with regard to their calcium-dependence and discuss their potential impact on synaptic function. PMID- 15820397 TI - Masters or slaves? Vesicle release machinery and the regulation of presynaptic calcium channels. AB - Calcium entry through presynaptic voltage-gated calcium channels is essential for neurotransmitter release. The two major types of presynaptic calcium channels contain a synaptic protein interaction site that physically interacts with synaptic vesicle release proteins. This is thought to tighten the coupling between the sources of calcium entry and the neurotransmitter release machinery. Conversely, the binding of synaptic proteins to presynaptic calcium channels regulates calcium channel activity. Hence, presynaptic calcium channels act not only as the masters of the synaptic release process, but also as key targets for feedback inhibition. PMID- 15820398 TI - Presynaptic Ca2+ dynamics, Ca2+ buffers and synaptic efficacy. AB - In synapses neurotransmitter release is triggered by elevation of Ca2+ concentration at a Ca2+ sensor of the release machinery. The Ca2+ concentration at the release site at the given time point is determined by Ca2+ dynamics within presynaptic terminal. It depends on a source of Ca2+ (usually voltage-gated Ca2+ channels), diffusional distance between the source of Ca2+ and the Ca2+ sensor and Ca2+ buffering by endogenous Ca2+ buffers. In many synapses transmitter release can be enhanced (facilitated) during repetitive activity of neurons. The main source of facilitation is activity-dependent increase of Ca2+ concentration at the release site. Several mechanisms of facilitation have been proposed, namely, accumulation of residual Ca2+, multi-site (X receptor) mechanism and partial Ca2+ buffer saturation mechanism. In this review we discuss theoretical and experimental evidence in favor of one or the other of proposed mechanisms. PMID- 15820399 TI - Calcium and endocannabinoids in the modulation of inhibitory synaptic transmission. AB - Synapses in the central nervous system can be highly plastic devices, being able to modify their efficacy in relaying information in response to several factors. Calcium ions are often fundamental in triggering synaptic plasticity. Here, we will shortly review the effects induced by postsynaptic increases of calcium concentration at GABAergic and glycinergic synapses. Both postsynaptic and presynaptic mechanisms mediating changes in synaptic strength will be examined. Particular attention will be devoted to phenomena of retrograde signaling and, specifically, to the recently discovered role, played by the endocannabinoid system in retrograde synaptic modulation. PMID- 15820400 TI - Dynamic aspects of presynaptic calcium currents mediating synaptic transmission. AB - Ca2+ entry through voltage-gated Ca2+ channels (VGCC) triggers transmitter release. Direct recording of Ca2+ currents from the calyx of Held nerve terminal revealed that presynaptic VGCCs undergo various modulations via presynaptic G protein-coupled receptors (GPCRs), Ca2+-binding proteins and a developmental switch of their alpha1 subunits. Dynamic changes of presynaptic VGCCs alter synaptic efficacy, thereby contributing to a variety of modulations of the CNS function. PMID- 15820402 TI - Lipidic carriers of RNA/DNA oligonucleotides and polynucleotides: what a difference a formulation makes! PMID- 15820403 TI - Role of polyanhydrides as localized drug carriers. AB - Many drugs that are administered in an unmodified form by conventional systemic routes fail to reach target organs in an effective concentration, or are not effective over a length of time due to a facile metabolism. Various types of targeting delivery systems and devices have been tried over a long period of time to overcome these problems. Targeted delivery or localized drug delivery offers an advantage of reduced body burden and systemic toxicity of the drugs, especially useful for highly toxic drugs like anticancer agents. Local drug delivery via polymer is a simple approach and hypothesized to avoid the above stated problems. Polyanhydrides are a unique class of polymer for drug delivery because some of them demonstrate a near zero order drug release and relatively rapid biodegradation in vivo. Further, the release rate of polyanhydride fabricated device can be altered over a thousand fold by simple changes in the polymer backbone. Hence, these are one of the best-suited polymers for drug delivery, with biodegradability and biocompatibility. The review focuses on the advantages of polyanhydride carriers in localized drug delivery along with their degradability behavior, toxicological profile and role in various disease conditions. PMID- 15820404 TI - Novel powder formulations for controlled delivery of poorly soluble anticancer drug: application and investigation of TPGS and PEG in spray-dried particulate system. AB - Biodegradable poly (lactic-co-glycolic acid) (PLGA), D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS) and/or polyethylene glycol (PEG) were combined as pharmaceutical excipient to fabricate microparticles containing sparingly soluble drug paclitaxel by spray-drying technique with successful achievement. The effect of formulation variety on particle morphology, surface composition, thermal property, drug entrapped capability, and drug release profile was investigated. The result indicated that the use of the appropriate mixtures of PLGA, TPGS and/or PEG produced paclitaxel-loaded microparticles characterised by acceptable pharmaceutical properties. Atomic force microcopy (AFM) and scanning electron microscopy (SEM) showed that the produced microparticles were spherical in shape with dimples or pores. The particle size ranged from 0.88 to 2.44 microm with narrow distribution. The combination of TPGS and PEG in the formulation resulted in a narrow particle size distribution in general although the influence of the formulation on the particle size was not significant. Differential scanning calorimetry (DSC) study implied that all those components in consideration were compatible well in the blend formulation systems. The paclitaxel entrapped in the particles existed in an amorphous or disordered-crystalline status in the matrices and was independent of the PLGA/TPGS/PEG ratio. X-ray photoelectron spectroscope (XPS) analysis revealed that after incorporation the particle's surface was dominated with PLGA due to its hydrophobic property. The formulation variety had an important impact on the drug release that was reduced with the presence of large fraction of TPGS resulting from a strong hydrophobic interaction between various matrix materials and the drug inside the particle. A zero order release could be yielded by optimising the ratio of PLGA/TPGS/PEG. The combination of PLGA/TPGS/PEG as safe pharmaceutical excipient to formulate particulate delivery system is beneficial in improving the pharmaceutical properties for further powder dosage application. PMID- 15820405 TI - Synergistic enhancement of testosterone transdermal delivery. AB - In this study, the effects of occlusion, octisalate (OS), and propylene glycol (PG) on the in vitro skin permeability of testosterone (TES) have been investigated. TES (either alone or with OS 5% w/v) was applied as a finite dose to full-thickness neonatal porcine skin mounted in flow-through diffusion cells and the amount of TES appearing in the receptor solution (20% v/v ethanol) was determined over 24 h. The skin was occluded with a microscope glass cover slip and to determine the effect of PG, 400 microl of PG/water mixtures (of varying PG concentration) was applied. In addition, the effect of Solugel (a proprietary hydrogel containing PG 25% w/w) and Tegaderm (a semipermeable film dressing) on the permeation of TES was assessed. Occlusion had no effect on the permeation of TES, however, OS increased the flux of TES 2.9-fold. The concentration of PG which produced optimal TES flux was 20% v/v, and this concentration resulted in a 1.9-fold increase in TES permeation. By combining OS, PG, and occlusion, TES permeation was increased 8.7-fold, which was a synergistic enhancement. In addition, Solugel and Tegaderm, when applied to the skin, produced a similar enhancement in TES permeation to that produced by PG 25% w/w and occlusion. PMID- 15820406 TI - Elucidation of the transport pathway in hairless rat skin enhanced by low frequency sonophoresis based on the solute-water transport relationship and confocal microscopy. AB - In this study, we examined a relationship between hydrophilic solute and water (vehicle) transports in the excised hairless rat skin in the presence of ultrasound (41 kHz, 60-300 mW/cm2) irradiation and also conducted skin surface observation using confocal microscopy. When the applied intensity was increased stepwise over the rage of 60-300 mW/cm2, the transport of tritiated water (3H2O) was increased 140-fold in an intensity-dependent manner and this returned to normal on stopping the ultrasound application. The skin permeation clearance (mul/h) of model hydrophilic solutes, calcein (MW 623) and FITC-labeled dextrans [MW 4400 (FD-4) and MW 38000 (FD-40)], across the skin under the influence of ultrasound was plotted against the corresponding 3H2O flux (microl/h) to estimate the potential contribution of convective solvent flow, induced by the ultrasound application, to the solute transport. Good correlations were observed between the 3H2O flux and solute clearances and, unexpectedly, the slope values obtained from linear regression of the plots were consistent for all solutes examined (1.04+/ 0.29 for calcein, 1.07+/-0.17 for FD-4, and 1.08+/-0.23 for FD-40, respectively). Transport of intact FD-4 and FD-40 was confirmed by gel permeation chromatography. When the skin surface and deeper regions of the skin after sonophoresis of FD-40 were observed using a confocal microscope, the fluorescence of FD-40 was uniformly distributed in the area under the ultrasound horn and also evident in crack-like structures in the boundary of the horn. On the other hand, a hexagonal structure of horny cells in the stratum corneum (SC) observed by post staining with rhodamine B was fully conserved in the area under the horn. These findings suggest that 41 kHz ultrasound can increase the transdermal transport of hydrophilic solutes by inducing convective solvent flow probably via both corneocytes and SC lipids as well as newly developed routes. Our observation also suggests that 41 kHz (low-frequency) ultrasound has the potential to deliver hydrophilic large molecules transdermally. PMID- 15820407 TI - Etanidazole in pH-sensitive liposomes: design, characterization and in vitro/in vivo anti-Trypanosoma cruzi activity. AB - In this work, the hydrophilic, low molecular weight and trypanocidal drug etanidazole (ETZ) was loaded in pH-sensitive liposomes (L-ETZ). Liposomes were made of dioleoyl-phosphatidylethanolamine: cholesteryl hemisuccinate (DOPE:CHEMS, 6:4, mol:mol), of 380 nm size at 14% ETZ/total lipid (w/w) ratio. To follow their uptake and intracellular fate by fluorescence microscopy, pH-sensitive liposomes were loaded with the fluorophore/quencher pair HPTS/DPX. A fast and massive delivery of the liposomal aqueous content into the cytosol of murine J774 macrophages was observed. L-ETZ vesicles were phagocytosed by both uninfected and Trypanosoma cruzi-infected macrophages. A 72% of anti-amastigote activity (AA) was demonstrated on L-ETZ-treated J774 cells, whereas the same dose of free ETZ rendered 0% AA. Endovenous administration of L-ETZ at 14 microg/mouse dose provoked significant decrease in parasitemia levels of T. cruzi-infected mice. Conversely, inoculation of a 180-fold higher dose of free ETZ failed in reducing the number of bloodstream trypomastigotes. Hence, these results point to develop systems, such as L-ETZ, designed for selective delivery of drugs to the cytoplasm of phagocytic cells, thus enhancing the efficacy of molecules considered poorly active. PMID- 15820408 TI - PEG-grafted chitosan as an injectable thermosensitive hydrogel for sustained protein release. AB - Thermosensitive polymer hydrogels that undergo a sol-to-gel transition in response to temperature changes are of great interest in therapeutic delivery and tissue engineering as injectable depot systems. A chitosan-based, injectable thermogel was prepared by grafting an appropriate amount of PEG onto the chitosan backbone and studied for drug release in vitro using bovine serum albumin (BSA) as a model protein. When more than approximately 40 wt.% of PEG was grafted to chitosan chains via covalent bonding, the aqueous solution of the resultant copolymer was an injectable liquid at low temperature and transformed to a semisolid hydrogel at body temperature. After an initial burst release in the first 5 h, a steady linear release of protein from the hydrogel was achieved for a period of approximately 70 h. Prolonged quasi-linear release of protein up to 40 days was achieved by crosslinking the hydrogel with genipin in situ, in a fashion suitable for protein encapsulation while maintaining the injectability of the hydrogel. The crosslinkage transformed the copolymer from a physical gel to an insoluble chemical gel and substantially reduced the initial burst release of protein. Both high performance liquid chromatography (HPLC) and gel electrophoresis indicated that the primary structure of BSA released from the hydrogels with or without genipin-crosslinking was generally conserved. The hydrogel can be prepared in solutions with a physiological pH, allowing the safe incorporation of bioactive molecules for a broad range of medical applications, particularly for sustained in vivo drug release and tissue engineering. PMID- 15820409 TI - Folate receptor mediated intracellular protein delivery using PLL-PEG-FOL conjugate. AB - To develop a receptor-mediated intracellular delivery system that can transport therapeutic proteins or other bioactive macromolecules into a specific cell, a di block copolymer conjugate, poly(L-lysine)-poly(ethylene glycol)-folate (PLL-PEG FOL), was synthesized. The PLL-PEG-FOL conjugate was physically complexed with fluorescein isothiocyanate conjugated bovine serum albumin (FITC-BSA) in an aqueous phase by ionic interactions. Cellular uptake of PLL-PEG-FOL/FITC-BSA complexes was greatly enhanced against a folate receptor over-expressing cell line (KB cells) compared to a folate receptor deficient cell line (A549 cells). The presence of an excess amount of free folate (1 mM) in the medium inhibited the intracellular delivery of PLL-PEG-FOL/FITC-BSA complexes. This suggests that the enhanced cellular uptake of FITC-BSA by KB cells in a specific manner was attributed to folate receptor-mediated endocytosis of the complexes having folate moieties on the surface. The PLL-PEG-FOL di-block copolymer could be potentially applied for intracellular delivery of a wide range of other biological active agents that have negative charges on the surface. PMID- 15820410 TI - Starch/Carbopol spray-dried mixtures as excipients for oral sustained drug delivery. AB - The present study evaluated if mixtures prepared by spray-drying an aqueous dispersion of Amioca starch and Carbopol 974P could be used as matrix for oral sustained drug delivery. The influence of the Amioca/Carbopol 974P ratio (0/100, 25/75, 50/50, 60/40, 85/15, 90/10, 95/5 and 100/0) and the pH and ionic strength (mu) of the dissolution medium on the drug release was investigated. The matrices composed of the spray-dried mixtures with 10% or 15% Carbopol 974P sustained the drug release over the longest time period. At this Carbopol concentration, shear viscosity measurements indicated the formation of an optimal network between the polymer chains of Amioca starch and Carbopol 974P, forming a rigid gel layer offering resistance to erosion during the dissolution experiments. PMID- 15820411 TI - Trimethylated chitosans as non-viral gene delivery vectors: cytotoxicity and transfection efficiency. AB - Chitosans are linear polysaccharides of natural origin that show potential as carriers in drug and gene delivery. Introducing quaternisation on the chitosan backbone renders the polymer soluble over a wider pH range and confers controlled cationic character. This study aims to investigate the effect of increasing quaternisation and therefore, positive charge on cell viability and transfection. Oligomeric and polymeric chitosans were trimethylated, the toxicity and transfection efficiency of these derivatives were tested with respect to increasing degree of trimethylation. The cytoxicity of polymer and oligomer derivatives alone and of their complexes with plasmid DNA were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on COS-7 (monkey kidney fibroblasts) and MCF-7 (epithelial breast cancer) cells. Transfection efficiency was investigated using the pGL3 luciferase reporter gene on the same cell lines. Complexes were characterised for their stability by gel electrophoresis. Cytotoxicity results showed that all derivatives were significantly less toxic than linear polyethylenimine (PEI). A general trend of increasing toxicity with increasing degree of trimethylation was seen. However, higher toxicity was seen in polymeric chitosan derivatives over oligomeric chitosan derivatives at similar degrees of trimethylation. All derivatives complexed pGL3 luc plasmid DNA efficiently at 10:1 ratio and three (TMO44, TMC57 and TMC93) were able to transfect MCF-7 cells with greater efficiency than PEI; 16, 23 and 50-fold, respectively. TMC57, TMC93 and all TMOs gave appreciable transfection of COS-7 cells. PMID- 15820412 TI - Urinary excretion of biopyrrins, oxidative metabolites of bilirubin, increases in patients with psychiatric disorders. AB - Several authors have suggested that psychological stress induces the production of reactive oxygen species (ROS). Several studies have supported the idea that bilirubin exerts antioxidative effects in vivo, and it was reported psychological stress provokes bilirubin oxidation in vivo [Yamaguchi T., Shioji I., Sugimoto A., Yamaoka M., 2002. Psychological stress increases bilirubin metabolites in human urine. Biochem. and Biophys. Res. Commun. 293, 517-520]. We investigated whether the concentration of bilirubin oxidative metabolites (biopyrrins) is increased in urine from patients with psychiatric disorders. The concentration of biopyrrins in urine of 25 patients with psychiatric disorders (schizophrenia, 15; depression, 10) was compared with 96 healthy volunteers. The concentrations of biopyrrins, as measured by enzyme-linked immunosorbent assay, were normalized to the urinary concentration of creatinine. The concentration of biopyrrins in patients with psychiatric disorders (schizophrenia and depression) was significantly higher than that of healthy volunteers. In schizophrenia, biopyrrins levels correlated with scores of the Brief Psychiatric Rating Scale (BPRS), and in depression, biopyrrins levels correlated with scores of the Hamilton Depression Rating Scale (HAM-D). These finding suggest that psychotic states are associated with an increase in the oxidative metabolites of bilirubin in human urine. PMID- 15820413 TI - The involvement of dehydroepiandrosterone (DHEA) and its sulfate ester (DHEAS) in blocking the therapeutic effect of electroconvulsive shocks in an animal model of depression. AB - Depressed patients with resistance to electroconvulsive treatment (ECT) had high basal serum levels of dehydroepiandrosterone (DHEA) sulfate (DHEAS). To clarify the role of DHEA/S in the ECT resistance, Flinder Sensitive Line (FSL) rats, which are a genetic animal model of depression, were injected i.p. with 2 mg/kg DHEA daily for 13 days to overload their serum and brain DHEA/S levels. Thereafter, rats were exposed to electroconvulsive shock (ECS), which is analogue to ECT in humans. Both ECS and DHEA displayed an antidepressive-like effect, as assessed by immobility time in forced swim test. However, combined DHEA and ECS treatment abolished these antidepressive-like effects. In addition, the levels of neurosteroids, corticosterone and adrenocorticotropin in selected brain regions were evaluated and compared to serum levels. The present study supports our assumption that high basal levels of DHEA/S play a role in the resistance to ECS and maybe ECT in humans. PMID- 15820414 TI - Pharmacokinetic comparison of two methods of heroin smoking: 'chasing the dragon' versus the use of a heating device. AB - In preparation for a trial on co-prescription of inhalable heroin and methadone, two methods for inhalation of heroin/caffeine tablets were compared: the commonly used method of 'chasing the dragon' and a standardised procedure for inhalation of volatilised heroin, using a heating device. Five male addicts inhaled a tablet of smokable heroin daily for 5 days, alternating the inhalation method. Plasma concentrations of heroin, 6-acetylmorphine, morphine and morphine-3- and -6 glucuronide were determined using a liquid chromatography method with tandem mass spectrometric detection. The exposure to heroin and its metabolites (expressed as areas under the concentration-time curve) was significantly lower after smoking via the heating device than after 'chasing the dragon': heroin 80% and 6 acetylmorphine 73% lower (p < 0.05). Maximal concentrations of heroin and 6 acetylmorphine were also 80% and 70% lower (p < 0.05) after using the heating device. 'Chasing the dragon' is a more efficient inhalation method than inhalation via the heating device. PMID- 15820415 TI - Haloperidol treatments increased macrophage activity in male and female rats: influence of corticosterone and prolactin serum levels. AB - Haloperidol is a receptor D2 antagonist frequently used in the treatment of schizophrenic patients. Haloperidol increased prolactin release from anterior pituitary gland, and prolactin modulates immune system activity. Groups of six male and female rats received an acute 2 mg/kg haloperidol treatment (E1), or a long-term (E2) haloperidol treatments (2 mg/kg/day for 21 days); control rats were treated similarly, but with control solution (groups C1 and C2, respectively). In this work long-term haloperidol treatment (E2) increased macrophage spreading, phagocytosis and NO release in male and female rats. However, acute haloperidol treatment (E1) did not change macrophage activity. Corticosterone and prolactin serum levels were increased after acute (E1) and long-term (E2) haloperidol treatments in male and female rats, being this increment higher in female. Macrophage of male and female rats presented the same pattern of alterations after acute and long-term haloperidol treatments. Haloperidol-induced macrophage activation was discussed in the light of a possible indirect effect through prolactin increments in rats, or, alternatively, as a consequence of a direct action of macrophage dopamine receptor. PMID- 15820416 TI - Anxiogenic effects of Sumatriptan in panic disorder: a double-blind, placebo controlled study. AB - BACKGROUND: Several lines of evidence point to serotonergic abnormalities in patients with panic disorder (PD). Our goal was to further examine central serotonergic function in panic patients using autonomic and subjective responses to the postsynaptic serotonin 5-HT1D receptor agonist Sumatriptan. METHOD: Using a double-blind, randomized, placebo-controlled design, we assessed autonomic and subjective responses to oral Sumatriptan (100 mg) and placebo in 15 patients with PD, free of medication. Subjective responses were measured using the Hamilton Anxiety Rating Scale (HAM-A), National Institute of Mental Health Anxiety Scale (NIMHA), a modified version of the Panic Symptom Inventory (PI), Hamilton Depression Rating Scale (HAM-D), and Montgomery-Asberg Depression Rating Scale (MADRS). RESULTS: PD patients exhibited significantly enhanced autonomic and subjective responses following challenge with Sumatriptan. We observed an increased pulse rate and augmentation of various parameters measured on different anxiety scales. A constant inclination of aggravation of the measured parameters was detected during the hour post challenge. CONCLUSION: Oral administration of Sumatriptan, a 5-HT1D agonist, has been associated with an anxiogenic effect in PD patients. PMID- 15820417 TI - Chronic administration of the SSRI fluvoxamine markedly and selectively reduces the sensitivity of cortical serotonergic neurons to footshock stress. AB - We have evaluated, with the use of vertical microdialysis, the effects of fluvoxamine, a selective serotonin reuptake inhibitor (SSRI) on the increase in serotonin and norepinephrine output elicited in rats prefrontal cortex by exposure to footshock stress. Exposure to footshock stress induced a marked increase in the cortical extracellular concentration of both serotonin and norepinephrine (+70% and +100%, respectively) in control rats. Long term, but not acute administration of fluvoxamine (10 mg/kg, i.p. once a days for 21 days) completely antagonized the stress induced increase in cortical serotonin extracellular concentration, while failed to modify the sensitivity of cortical noradrenergic neurons to the same stressful stimulus. Our results have shown that it is possible to independently modulate the sensitivity of cortical serotonergic neurons to stressful stimuli without altering the responsiveness of noradrenergic neurons to the same stress. Given the different role played by serotonin and norepinephrine in the modulation of the stress response, the availability of drugs able to selectively modulate the plastic response of serotonergic neurons to stress in specific brain areas might be important for the pharmacotherapy of anxiety disorders. PMID- 15820418 TI - Nutritional and life style determinants of plasma homocysteine in schizophrenia patients. AB - Recently, homocysteine levels have been reported to be elevated in young male schizophrenic patients. Since smoking, obesity, low folate or low vitamin B12 and various medications can increase homocysteine levels, we studied these variables and other clinical variables in 258 schizophrenic patients. A multiple linear regression for plasma homocysteine was performed on variables that were significantly related to plasma homocysteine. Variables predicting homocysteine levels in schizophrenic patients include gender, plasma folate levels, plasma vitamin B12 levels, mean red blood cell corpuscular volume and diastolic blood pressure. Only 24% of the variance in male patients was explained by the model. The reason for elevated plasma homocysteine in some schizophrenic populations remains unclear. PMID- 15820419 TI - Reinstatement of cocaine-seeking behavior in rats is attenuated following repeated treatment with the opioid receptor antagonist naltrexone. AB - In the present study we show that the endogenous opioid systems play a modulating role in cocaine-induced reinstatement of drug-seeking behavior in rats. We investigated the effect of blockade of opioid receptors on reinstatement of cocaine-seeking behavior by cocaine priming. Drug-naive rats were allowed to initiate self-administration behavior of cocaine (30 and 60 mug per infusion, i.v.) for 5 consecutive daily sessions, and after a 5-day extinction period during which the rats did not receive cocaine, a test for cocaine-induced (1 mg/kg, i.v.) reinstatement followed. The effect of cocaine priming was tested on days 1, 3, and 5 after extinction, while on days 2 and 4 the animals received saline priming. Before each daily reinstatement test, different groups of animals received an injection with the opioid receptor antagonist naltrexone (3 mg/kg, s.c.) or with placebo. We observed that cocaine readily reinstated extinguished responding in the rats, and that this reinstatement responding did not change over the consecutive reinstatement tests. Pretreatment with naltrexone progressively attenuates the cocaine-induced reinstatement, with a significant reduction on days 3 and 5 of reinstatement testing. Discriminative lever-pressing (active versus inactive lever) during reinstatement phase, however, remains present in animals treated with naltrexone. This implies that repeated opioid receptor blockade progressively attenuates cocaine-induced drug-seeking behavior in abstained animals, but this cannot simply be attributed to extinction of cocaine-seeking behavior. PMID- 15820420 TI - Lowered DHEA-S plasma levels in adult individuals with autistic disorder. AB - The aim of this study was to determine for the first time neurosteroid levels, dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEA-S) in particular, in a group of adult patients with autistic disorder and compare these levels with normal healthy individuals. Levels of DHEA, DHEA-S and cortisol were compared between 15 adult drug-free patients with autistic disorder and 13 healthy controls. The Ritvo-Freeman Real-Life Rating Scale (RLRS) and the Overt Aggression Scale (OAS) were assessed as a measure of symptom severity. Significant lower DHEA-S levels were observed in the group with autistic disorder as compared to controls (p < 0.05). DHEA-S levels appear to be low in patients with autistic disorder and, while speculative, may play a role in the etiopathophysiology of the disorder. PMID- 15820421 TI - Effect of chronic olanzapine treatment on nerve growth factor and brain-derived neurotrophic factor in the rat brain. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are proteins involved in neuronal survival, neurite outgrowth and synapse formation. Recent observations suggest that treatment with typical and atypical antipsychotic drugs affect NGF and BDNF levels in the rat brain. The atypical antipsychotic olanzapine has a low incidence of side effects, such as extrapyramidal and anticholinergic symptoms. Since NGF and BDNF are involved in the regulation of cholinergic, dopaminergic and serotonergic neurons in the central nervous system (CNS) we hypothesized that chronic olanzapine treatment will influence the distribution of NGF and BDNF in the rat brain. To test this hypothesis we administered olanzapine for 29 days in the drinking water at the doses of 3 and 15 mg/kg body weight and measured the levels of NGF and BDNF in the brain of Wistar rats. Olanzapine increased NGF in the hippocampus, occipital cortex and hypothalamus. In contrast, olanzapine decreased BDNF in the hippocampus and frontal cortex. Although the significance of these findings is not clear, a heuristic hypothesis is that olanzapine's clinical effects and a favorable side effect profile are in part mediated by neurotrophins. PMID- 15820422 TI - The possible role of neurotrophins in the pathogenesis and therapy of schizophrenia. AB - The pathogenesis of schizophrenia may be ascribed to early maldevelopment of brain tissue. Neurotrophins are a group of dimeric proteins that affect the development of the nervous system in all vertebrates' species. Since neurotrophins, as well as other growth factors, play a crucial role in neurodevelopment, they are plausible candidates of taking part in the pathophysiology of schizophrenia. In line with this hypothesis, accumulating preclinical and clinical data indicate that dysfunctions of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) may contribute to impaired brain development, neuroplasticity and synaptic "dysconnectivity" leading to the schizophrenic syndrome, or at least some of its presentations. This article reviews the functions of neurotrophins in the complex process of normal brain development, and their possible relevance to the neuropathology and neuropharmacology of schizophrenia. Further research in this area may bring about novel pharmacological therapeutic strategies to this chronic debilitating disorder. PMID- 15820423 TI - Rats with scopolamine- or MK-801-induced spatial discrimination deficits in the cone field task: animal models for impaired spatial orientation performance. AB - Spatial cognition appears to be compromised in elderly and in patients suffering from dementia. These deficits are believed to be modelled, at least partly, by the administration of scopolamine or MK-801 in normal adult animals. In order to establish an animal model suited for the evaluation of putative cognition enhancers, we assessed the effects of scopolamine (0.3, 0.5, 0.7 mg kg(-1), i.p.) and MK-801 (0.07, 0.08, 0.09 mg kg(-1), s.c.) in rats trained in the cone field. This task allows the simultaneous investigation of working memory (WM), reference memory (RM) and search strategies. Scopolamine and MK-801 reliably induced spatial cognition deficits in the cone field without inducing behavioural side effects. This task appears to be suited for assessing the effects of putative cognition-enhancing compounds on spatial cognition. PMID- 15820426 TI - A computer-assisted approach to planning multidimensional distraction osteogenesis. PMID- 15820424 TI - Possible association between the gamma-aminobutyric acid type B receptor 1 (GABBR1) gene and schizophrenia. AB - Schizophrenia (SCZ) is a severe neuropsychiatric disorder with a genetic component. The major inhibitory GABA-(gamma-aminobutyric acid) ergic system may be involved. The GABA type B receptor 1 (GABBR1) gene has been localized to 6p21.3, a region linked to SCZ. We therefore investigated five polymorphisms (A 7265G, C10497G, Ser-491-Ser-T1473C, Phe-659-Phe-T1977C, and 3'-UTR A33795G substitutions) in the GABBR1 gene in a sample of 101 DSM-IV SCZ probands and their families, 150 unrelated affected individuals matched with 150 healthy controls, using the transmission disequilibrium test (TDT) and case-control analysis. We did not observe biased transmission of alleles in any of the polymorphisms individually and haplotypes within the gene to SCZ probands. However, a weak significant difference was observed in the A-7265G polymorphism between the allelic frequency (chi2 = 4.310, P = 0.038) and a trend was observed between the genotype frequency (chi2 = 4.970, 2 df, P = 0.083) of SCZ individuals and controls. Further investigations of the role of GABBR1 in SCZ are warranted. PMID- 15820427 TI - Geometric considerations in the transition from two-dimensional to three dimensional treatment planning. PMID- 15820428 TI - Three-dimensional computer-aided surgical simulation for maxillofacial surgery. PMID- 15820429 TI - Computer-assisted navigational surgery in oral and maxillofacial surgery. PMID- 15820430 TI - Imaging-guided biopsy. PMID- 15820431 TI - Ultrasound evaluation of bone healing in distraction osteogenesis of the mandible. PMID- 15820432 TI - Interactive CT software in oral and maxillofacial surgery. PMID- 15820433 TI - Custom-made total temporomandibular joint prostheses. PMID- 15820434 TI - [New strategies to interfere with radiation response: "biomodulation" of radiation therapy]. AB - The development of several new anti cancer agents has been made possible because of recent significant achievements in our global understanding of cancer biology. These new "targeted" agents selectively inhibit targets necessary for tumor cell growth and viability with little toxicity to normal cells compared to conventional cytotoxic agents. So far, the efficacy of many of these new promising agents when used alone treatment remains limited, it is likely that the optimal use of these agents could be obtained in combination with conventional agents such as radiation therapy. The potential benefit of these targeted therapies combined with irradiation seems important. They might offer the advantage of increasing the tumor response to radiation with no or little increase in normal tissue damage. Therefore, these new types of chemo-radiation approaches might respect the normal tissue versus tumor cell "therapeutic ratio". These approaches can be sub divided in three sub groups: 1) Therapeutics targeting selectively one tumor related biochemical activity such as EGFR inhibitors. These approaches are efficient but one mutation of the target might render them inefficient. 2) Therapeutics directed against a widely expressed target. This is the case for anti Insulin Growth Factor-1 (IGF1R) interventions: IGF1R inhibition seems to specifically alter tumor cell viability with a minimal effect on normal cells viability. 3) Strategies which are not targeted against the tumor but the microenvironment, especially angiogenesis. This type of approaches seems to be applicable independently of tumor intrinsic biologic related factors. PMID- 15820435 TI - [Genetic bases of the radiosensitivity of breast cancer]. AB - Local-regional radiation therapy is one of the major therapeutic means in the management of breast cancer. Three questions however arise from the important advances achieved in this domain in the past years. The first question concerns the possibilities to identify and overcome the radioresistance of a subset of tumours. The second question is how to recognize women likely to benefit from adjuvant radiation therapy, and therefore to diminish treatment indications in other groups. Finally, the third question is how to identify subjects at high risk for long term injury following breast irradiation, in order to adapt techniques and indications in such populations. The major advances of breast cancer molecular genetics in the past years should provide clinicians with tools to answer these important questions. In this paper, we review the molecular germline (BRCA1, BRCA2, ATM, ...) and somatic (p53, tyrosine kinase receptors, as well as actors of cell cycle, signal transduction, apoptosis, DNA repair ...) main bases of breast cancer radiosensitivity. Recent methods of exploration of the genetic background of both the host and the tumours (gene and protein expression profiles) are also reviewed as major tools of breast cancer management in the next few years. PMID- 15820436 TI - [Anemia impact on treatments of cervical carcinomas]. AB - During the treatments of carcinomas of the cervix, anemia is relatively frequent and its origin is complex combining often hemorrhage, iron deprivation, inflammatory reactions and infection. The frequency of the primary anemia (hemoglobin level<12 g/dl) is correlated with clinical stage and varies from one publication to another, mainly from 25% for stage I, to 33% for stage II and can approach 40% for stage III. Anemia is correlated with patient survival and it appears to be one of the most powerful prognostic factor after clinical stage and tumor size. Anemia is a bad prognostic factor related to stage and tumor size but it has not been proven to be an independent factor. Anemia increases hypoxia of cervix carcinomas, which is an independent prognostic factor for patients N0. Moreover, we know that the oxygenation of these tumors is correlated with hemoglobin levels. The normalization of Hb levels by transfusion could certainly modify the prognosis of patients anemic before treatment, or of those becoming anemic during radiotherapy treatment. For smokers, anemia is certainly more important that we can appreciate from the Hb levels only, by the presence of carboxyhemoglobin. Concomitant chemotherapies with cisplatin compounds are actually standards and they can largely increase the risk of inducing anemia, therefore more than 50% of patients will experiment it during their different treatments. Transfusion is recommended by the SOR (Standards Options and Recommendations of the Federation nationale des centres de lutte contre le cancer) under 10 g/dl. The use of erythropoietin is a therapeutic option for Hb levels between 10 and 12 g/dl and strongly recommended after a Hb normalization by blood transfusion. For 70% of patients who respond to erythropoietin, a better control of the Hb level is obtained. The impact of this anemia on quality of life and treatments compliance justifies the use of erythropoietin, especially in cancers for which treatments induce a deep fatigue and a very bad tolerance, which could be a limiting factor. PMID- 15820437 TI - [Radiotherapy-induced pemphigus: a case report]. AB - A 61-year-old male patient suffering from squamous cell carcinoma of the lower lip developed pemphigus vulgaris two months after exposure to radiotherapy. Skin lesions were initially localised to the face and neck and later extended over other skin areas. The eruption are improved with glucosteroid therapy, which were stopped after six months. Pemphigus induced by radiotherapy is rare, latency before the onset of the vesiculobullous eruption is variable. Clinical, histological and immunological characteristics are similar to those of other types of pemphigus. PMID- 15820438 TI - [Malignant melanoma of nasal fossae: clinical and therapeutic considerations about three cases]. AB - We report in this publication three cases of mucous malignant melanoma of the nasal fossae. The nasal mucous malignant melanoma creates a unilateral obstructive and hemorrhagic tumour, readily septal or situated on the external wall of the nasal fossae (inferior and middle turbinates). The diagnosis established by the clinical examination (pigmented forms) is confirmed by the anatomopathological examination--thanks in particular to its tinctorial affinity for Fontana-Masson colouring. The immunohistochemistry reinforces the diagnosis in the presence of any achromic varieties or those simulating an undifferentiated tumour or neurinoma. Modern imagery (CT Scan, IRM) is impossible to circumvent for a precise loco-regional assessment, an essential preliminary to the therapeutic protocol--mainly of radiosurgical type. Chemotherapy (deticene) remains in force in the metastatic forms. Specific therapies are being evaluated (immunological-hormonal therapy). The forecast of the mucous malignant melanoma remains pejorative with a 10 to 40% survival rate of five years. The improvement of the forecast depends on an early diagnosis, an evolutionary locoregional assessment as complete as possible with an adapted and multidisciplinary choice of therapies (surgery, radiotherapy, immunotherapy, hormonal therapy). PMID- 15820439 TI - [Minutes of the 23th meeting of the European Society for Therapeutic Radiology and Oncology (ESTRO). Amsterdam, 24-28 October 2004]. PMID- 15820440 TI - Different antioxidative potencies of dihydropyridine calcium channel modulators in various models. AB - There is evidence that dihydropyridine calcium antagonists (DHP) play a beneficial role during the development of atherosclerosis. Since antioxidative properties of this substance class may be important, we investigated the antioxidative potency of the DHP prototype calcium channel antagonist nifedipine, the long acting calcium channel antagonist lacidipine, the DHP calcium channel agonist Bay K 8644 and the bulky DHP derivate Bay O 5572 (negligible effects on L type calcium channels) in three different models. Additionally, we examined the potential correlation between lipophilic and antioxidative properties. In an in vitro model, Bay K 8644 was significantly more effective in scavenging superoxide anions (hypoxanthine/xanthine-oxidase-assay) than lacidipine, Bay O 5572 or nifedipine (micro- to millimolar concentration range). Addition of artificial membrane preparations (dimyristoylphosphatidylcholine) to mimic a physiological environment resulted in an enhanced antioxidative effect, with lacidipine being the most effective DHP to quench radicals (low micromolar concentration range). Thirdly, in a more physiological model of hyperglycemia (30 mmol/l) induced release of reactive oxygen species (ROS) from native endothelial cells of porcine coronary arteries, we showed that nifedipine was a significantly more potent antioxidant (therapeutical nanomolar concentration range) than the other DHP. Calculation of the lipophilicity of the four substances (lacidipine>Bay O 5572>Bay K 8644>nifedipine) showed a positive correlation between the antioxidative potency and the lipophilicity in the model with the artificial membranes but not in the other models. We conclude that it seems necessary to access antioxidative properties of substances in physiological models in which we could demonstrate that nifedipine exhibits ROS-quenching properties in a therapeutic concentration range. PMID- 15820442 TI - Rapid and simple determination of epoxyeicosatrienoic acids in rabbit renal artery by reversed-phase HPLC with fluorescence detection. AB - A liquid chromatographic method with fluorescence detection coupled with a solid phase extraction was applied to the rapid determination of epoxyeicosatrienoic acids (EETs) in the rabbit renal artery. The EETs were extracted with an acetonitrile from renal artery homogenate and concentrated by a solid-phase extraction method. The concentrated EETs were reacted directly with a 6, 7 dimethoxy-1-methyl-2 (1H)-quinoxalinone-3-propionyl-carboxylic acid (DMEQ) hydrazide and separated by a reversed-phase HPLC with eluting a combination of a step-wise and a gradient of a mixture of methanol and water. The content of EETs in the renal arteries was significantly greater in the 0.5% cholesterol fed rabbits than in control rabbits. It is suggested that hyperchlesterolemia increases the production of EETs in the rabbit renal artery. PMID- 15820441 TI - Isoform-specific modulation of coronary artery PKC by glucocorticoids. AB - Glucocorticoids (GC) exert diverse cellular effects in response to both acute and chronic stress, the functional consequences of which have been implicated in the development of cardiovascular pathology such as hypertension and atherosclerosis. However, the mechanisms by which GCs activate divergent signaling pathways are poorly understood. The present study examined the direct effects of natural (cortisol) and synthetic (dexamethasone) GCs on protein kinase C (PKC) isoform expression in coronary arteries. Porcine right coronary arteries were treated in vitro for 18 h in the presence and absence of either dexamethasone (10, 100, or 500 nM) or cortisol (50, 125, 250, or 500 nM). PKC isoform levels and subcellular distribution were determined by immmunoblotting of whole cell homogenates and immunocytofluorescence using PKC-alpha, -betaII, -epsilon, -delta, and -zeta specific antibodies. Dexamethasone caused a approximately 4-fold increase in PKC alpha, a approximately 2.5-fold increase in PKC-betaII, and a 2-fold increase in PKC-epsilon (p<0.05). In contrast, dexamethasone had no effect on PKC-delta or PKC- zeta levels. Dexamethasone also caused an increase in the activity of PKC alpha (285%), -betaII (170%), and -epsilon (210%). Cortisol produced similar effects on PKC isoform expression. Confocal microscopy revealed that while dexamethasone altered localization patterns for PKC-alpha, -betaII and -epsilon, no such effect was observed for PKC-delta or PKC-zeta. The stimulatory effects of dexamethasone and cortisol on coronary PKC levels and translocation were prevented by the GC receptor (GR) blocker, RU486. These results demonstrate, for the first time, that GCs modulate coronary PKC expression and subcellular distribution in an isoform-specific manner through a GR-dependent mechanism. PMID- 15820443 TI - Inhaled nitric oxide improves the survival of the paraquat-injured rats. AB - The purpose of this study was to evaluate the effects of inhaled nitric oxide (NO) on the paraquat-induced lung injury in rats. The rats were assigned to four groups: control; inhaled NO (5 ppm); paraquat (PQ, 30 mg/kg); and PQ+NO group. For first 18 h the inhalation of NO mixed with room air was performed. Total white blood cell (WBC), neutrophil, total protein, lactate dehydrogenase (LDH), transforming growth factor-beta1 (TGF-beta1) in serum and/or bronchoalveolar lavage (BAL) fluid, serum malonaldehyde (MDA), and myeloperoxidase (MPO) of lung were measured and lung histopathology were also reviewed. The 72-h survival rate of PQ group was 58%, but the survival rate of PQ+NO group, NO group and control group were 100%, respectively. The serum MDA and TGF-beta1 in BAL fluid and blood of PQ+NO group were significantly lower than those of PQ group. However, inhaled NO did not decrease the elevated total WBC and neutrophil counts, and total protein, LDH and MPO activity in the lung injured by PQ. The alveolar septal thickening and inflammatory cell infiltration were not different between PQ and PQ+NO groups. NO inhalation may be beneficial for the survival of paraquat induced injured rats by attenuating lipid peroxidation and production of TGF beta1. PMID- 15820444 TI - Evaluation of antioxidant effect of Semecarpus anacardium Linn. nut extract on the components of immune system in adjuvant arthritis. AB - The effect of Semecarpus anacardium Linn. nut extract (SA) on the level of Lipid peroxides (LPO) and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) in the lymphocytes and lymphoid organs, namely spleen and thymus of adjuvant induced arthritic rats, were investigated. The results were compared with normal and untreated arthritic rats. The levels of reactive oxygen species (ROS), namely Hydroxy radical, Superoxide radical, and H2O2 were also measured in spleen, thymus, and lymphocytes of control and experimental animals. Biochemical markers of inflammation namely C-reactive protein (CRP) level and Erythrocyte sedimentation rate (ESR) were determined. Anti-arthritic profile was evaluated from the changes in the paw edema and arthritic scores of arthritic and drug-treated rats. A significant increase in the level of LPO, ROS and decreased levels of antioxidant enzymes in arthritic rats were observed. On treatment with the drug, the above changes were reverted back to near normal levels. The increment in CRP level and ESR observed in arthritic animals were found to be significantly restored in SA treated rats. There were no significant changes in sole drug-administered normal rats. Semecarpus anacardium Linn. nut extract significantly decreased the paw edema and arthritic score in arthritic rats on administration, whereas in untreated arthritic rats, there was a significant edema in the hind paw. PMID- 15820445 TI - Histamine improves survival and protects against interleukin-2-induced pulmonary vascular leak syndrome in mice. AB - The therapeutic efficacy in the treatment of metastatic cancer with high doses of interleukin-2 (IL-2) has been limited by the onset of vascular leak syndrome (VLS) and related toxicities. VLS is characterized by an increase in vascular permeability and severe hypotension resulting in interstitial edema and organ failure. This study explores the protective effects of histamine dihydrochloride (HDC) against IL-2-induced toxicities in mice. Treatment with HDC administered before or after IL-2 (1.25 x 10(6) IU, BID) was shown to protect mice from VLS related toxicities and mortality in a dose-dependent manner. Survival rates when HDC was added were 56, 75 and 81% at doses of 0.47, 4.7 and 47.0 mg/kg, respectively, compared to 42% survival with IL-2 alone. HDC protected against IL 2-induced macroscopic pulmonary lesions, reduced edema (up to 62% reduction in lung wet/dry weight ratio) and reduced capillary leakage into the lungs as measured by a reduction in Evans Blue dye content. In addition, the systemic effect on serum cytokine levels showed that HDC only moderately lowered IL-2 induced IFN-gamma, IL-6, IL-10, IL-18 and TNF-alpha. Serum levels of IL-1beta, IL 4 and IL-12 were not measurably induced by IL-2 treatment. HDC modulates many cellular functions including regulating cytokines and blocking immune-suppression caused by reactive oxygen species (ROS) generated by the NADPH oxidase. However, the protective effect of HDC on alleviating IL-2-induced pulmonary edema was not related to ROS inhibition. Our data indicate that HDC treatment improves survival and protects against IL-2 induced VLS independent of ROS regulation in mice. PMID- 15820446 TI - A prospective study of outcomes of emergency and elective surgeries for complicated colonic cancer. AB - BACKGROUND: Although a significantly decreased long-term survival has been observed in patients undergoing surgery for complicated colorectal tumors compared with uncomplicated ones, the role of radical oncologic surgery on emergency colonic cancer is not defined clearly. The aim of this study was to analyze the efficacy of a curative emergency surgery in terms of tumor recurrence and cancer-related survival compared with elective colonic surgery. METHODS: Between January 1996 and December 1998, all patients with colonic cancer deemed to have undergone a curative resection were considered for inclusion in this prospective study. Patients were classified into 2 groups: group 1, after emergency surgery for complicated colonic cancer, and group 2, patients undergoing elective surgery. The main end points were cancer-related survival and the probability of being free from recurrence at 3 years. RESULTS: Of the 266 patients included in the study, 59 patients (22.2%) were in group 1 and 207 patients (77.8%) were in group 2. Postoperative mortality was higher in group 1 (P=.0004). After patients were stratified by the tumor node metastasis system, differences between the groups with respect to overall survival of stage II tumors (P=.0728), the probability of being free from recurrence (P=.0827), and cancer-related survival (P=.1071) of stage III cancers did not reach statistical significance. Differences were observed for the overall survival in stage III tumors (P=.0007), and for the probability of being free from recurrence (P=.0011) and cancer-related survival (P=.0029) in stage II cancers. When patients with elective stage II tumors presenting 1 or more negative prognostic factor were compared with emergency patients affected by a stage II colonic cancer, no differences were observed. CONCLUSION: Curative surgeries for complicated colonic cancer are acceptable in emergency conditions. Cancer-related survival and recurrence in patients with complicated colonic cancers may approach that of elective surgery if a surgical treatment with radical oncologic criteria is performed. PMID- 15820447 TI - Comparison of one-stage resection and anastomosis of acute complete obstruction of left and right colon. AB - BACKGROUND: Although acute obstruction of the right colon is usually handled by primary anastomosis following resection, many surgeons are reluctant to offer one stage resection and anastomosis to patients with obstructive lesions of the left colon. The aim of the study is to compare the immediate result of one-stage resection and anastomosis for patients with acute complete obstruction of the right colon versus left colon. METHODS: From January 1986 to December 2003, 214 cases of acute colonic obstruction were managed with one-stage resection and anastomosis by a single surgeon. Eighty patients were operated on for obstructive lesions of the right colon, 71 of them for carcinoma of the colon. Operative mortality was 10% (8/80); all except 2 patients died of respiratory failure. There were 2 cases (2.5%) of anastomotic leakage. One hundred thirty-four patients were operated on for obstructive lesions of the left colon, 127 of them for carcinomas of the colon and rectum. Operative mortality was 1.5% (2/134); both patients died of metastasis from the colorectal cancer following surgery. There were 3 cases (2.3%) of anastomotic leakage. CONCLUSION: This experience suggests that an anastomosis can be performed as safely in patients with acute obstruction of the left colon as in those with acute obstruction of the right colon. Mortality following resection and anastomosis is actually lower in left than right colonic obstruction. Neither intraoperative irrigation nor routine subtotal colectomy was found to be necessary in patients with acute colonic obstruction. Intraoperative decompression should be considered in left and also right colonic obstruction prior to the anastomosis following colonic resection. PMID- 15820448 TI - Intraoperative magnetic resonance imaging ablation of hepatic tumors. AB - BACKGROUND: The use of hepatic ablation of tumors for both primary and secondary cancers has continued to increase at a significant rate. The most significant increase in the use of hepatic ablation has come from image-guided techniques with computed axial tomography and ultrasound. Limitations to targeting hepatic lesions by these techniques remain morbid obesity, abnormal hepatic parenchyma, and inability to visualize lesions without the use of intravenous contrast. In contrast, magnetic resonance imaging (MRI) has continued to provide a high contrast of soft tissue-to-lesion conspicuity without the need for intravenous dye. The recent development of open-configuration magnetic resonance scanners- which have allowed improved patient access, near real-time imaging, and more available MRI-compatible equipment--has opened up an entire new area of image guided surgical and interventional procedures. METHODS: The principles and indications for all types of image-guided hepatic ablations are described. RESULTS: The success and limitations of image-guided ablation techniques. CONCLUSIONS: Image-guided hepatic ablation represents a useful technique in managing hepatic tumors. Intraoperative MRI represents a new technique with initial success that has been limited to European centers. Further evaluation in United States centers has demonstrated intraoperative MRI to be useful for certain hepatic tumors that cannot be adequately visualized by ultrasound or computed axial tomography. A multidisciplinary approach involving a surgical oncologist and interventional radiologist remains integral to the short- and long term success of image-guided ablation. PMID- 15820449 TI - Antimicrobial prophylaxis for surgery: an advisory statement from the National Surgical Infection Prevention Project. AB - In January 2003, leadership of the Medicare National Surgical Infection Prevention Project hosted the Surgical Infection Prevention Guideline Writers Workgroup meeting. The objectives were to review areas of agreement among the published guidelines for surgical antimicrobial prophylaxis, to address inconsistencies, and to discuss issues not currently addressed. The participants included authors from most of the published North American guidelines for antimicrobial prophylaxis and several specialty colleges. The workgroup reviewed currently published guidelines for antimicrobial prophylaxis. Nominal group process was used to draft a consensus paper that was widely circulated for comment. The consensus positions of the workgroup include that infusion of the first antimicrobial dose should begin within 60 minutes before surgical incision and that prophylactic antimicrobial agents should be discontinued within 24 hours of the end of surgery. This advisory statement provides an overview of other issues related to antimicrobial prophylaxis including specific suggestions regarding antimicrobial selection. PMID- 15820450 TI - Laparoscopic adrenalectomy for malignancy. AB - The superiority of the minimally invasive approach to adrenal resections has been well documented for benign pathology. With technical advances and increased experience, surgeons have successfully performed laparoscopic adrenalectomies for metastatic and primary malignancies of the adrenal gland. The technique of laparoscopic adrenalectomy as it pertains to malignant lesions is presented. A review of the literature demonstrates the safety and efficacy of laparoscopic adrenalectomy for metastatic colorectal, lung, and renal tumors. For primary adrenal malignancies, radical resections can be effectively performed laparoscopically; however, continued long-term follow-up is needed to establish the minimally invasive technique as the preferred approach. PMID- 15820451 TI - Surgical skill is predicted by the ability to detect errors. AB - BACKGROUND: Objective analysis methods of surgical performance are now available so comparison between surgeons is available. One such method is by direct observation using the Objective Structured Assessment of Technical Skills (OSATS), but this is a time-consuming process; therefore, a simple screening tool for the ability to detect errors (previously validated) was analyzed and considered as a predictor of qualitative performance. METHODS: Thirty-eight volunteer surgeons were recruited to the skills laboratory to undertake 3 exercises. Two were bench-top surgical tasks that were scored using the global rating of the OSATS technique. The third task was the ability to detect simple errors in 22 synthetic models of common surgical procedures, some of which contained purposefully made errors. P<.05 was deemed to be statistically significant. RESULTS: The scores (interquartile ranges in parentheses) for the 3 sections were excision of sebaceous cyst=21 (19,24), closure of small bowel enterotomy=23 (21,27), and identification of errors=31 (27,34). Three scorers blinded to the operative models exhibited an interobserver reliability of .9 and .91 for the video tasks, respectively. Spearman's rank correlations between the error examination and performance on the 2 tasks were both statistically significant at .69 (cystectomy) and .54 (enterotomy). CONCLUSIONS: The ability to detect simple surgical errors is a predictor of technical skill and performance of bench tasks. What must be answered is whether the use of such models and principles can shorten the qualitative surgical learning curve. PMID- 15820452 TI - Assessing the teaching of technical skills. AB - BACKGROUND: There is a lack of structured instruments to assess how technical skills are taught. We aimed to develop a practical assessment tool that is easy to use and will assess the teaching of technical skills. METHODS: A 5-point Likert global rating scale was constructed. An experienced surgeon independently assessed each surgery using the assessment tool. Trainees also used the assessment tool after the surgery. RESULTS: Forty-two surgeries were assessed: 26 open and 16 laparoscopic surgeries. Interrater reliability between the independent assessor and trainees was performed using a kappa coefficient of .77 and a P value of less than .05. CONCLUSIONS: Our study shows that our assessment instrument has a potential ability to assess any taught technical task. We will continue and expand the study in surgery, and we aim to expand it further to medical specialties (eg, internal medicine, and so forth) that also teach technical tasks (eg, chest drain insertion, and so forth). PMID- 15820454 TI - Clinical and economic outcomes of oral linezolid versus intravenous vancomycin in the treatment of MRSA-complicated, lower-extremity skin and soft-tissue infections caused by methicillin-resistant Staphylococcus aureus. AB - BACKGROUND: Resistant bacteria often complicate the management of skin and soft tissue infections of the lower extremities. This open-label study compared oral linezolid and intravenous vancomycin for management of complicated skin and soft tissue infections caused by methicillin-resistant Staphylococcus aureus (MRSA). METHODS: Patients aged 18 years or older with proven MRSA-related complicated skin and soft-tissue infections requiring surgical intervention were randomized to receive oral linezolid (n=30) or intravenous vancomycin (n=30) for 7 to 21 days. Clinical and microbiological outcomes, duration of hospitalization and drug treatment, and outpatient charges were determined. RESULTS: Linezolid was associated with greater rates of clinical cure and improvement (P=.015), a 3-day shorter median length of stay (P=.003), and reduced outpatient charges (P<.001). Vancomycin therapy was associated with more treatment failures and subsequent lower-extremity amputations (P=.011). CONCLUSIONS: Clinical outcomes were significantly better with linezolid than with vancomycin. Additionally, linezolid was associated with reduced length of stay and outpatient charges. PMID- 15820453 TI - Should the inferior vena cava be reconstructed after resection for malignant tumors? AB - BACKGROUND: Caval replacement after circumferential resection of the inferior vena cava remains controversial. The aim of the current study is to determine whether or not inferior vena cava replacement should be performed. METHODS: We reviewed 36 cases undergoing resection of the inferior vena cava concomitant with resection of malignant neoplasms. Our criteria for circumferential resection of the inferior vena cava were half or more of the circumference of the vessel wall invaded by tumor, a primary tumor of the caval wall, or massive intraluminal tumor thrombus suspected of adhering to the caval wall. We detailed 10 patients undergoing circumferential resection of the inferior vena cava. RESULTS: Most of patients who did not undergo replacement of the inferior vena cava showed no sign of swelling of the lower limbs, but one showed persistent leg edema with oliguria. This patient had poor development of collateral circulation and mild obstruction of the inferior vena cava before surgery. Two patients who underwent replacement of inferior vena cava had no venous sequelae, although they had poor development of collateral circulation before surgery. CONCLUSION: Caval replacement after circumferential resection of the inferior vena cava may be necessary in patients who have preoperative poor development of collateral circulation or who have oliguria or unstable hemodynamics intraoperatively. PMID- 15820455 TI - Surgical versus chemical (botulinum toxin) sphincterotomy for chronic anal fissure: long-term results of a prospective randomized clinical and manometric study. AB - BACKGROUND: The aim of this prospective randomized trial was to compare the effectiveness and morbidity of surgical versus chemical sphincterotomy in the treatment of chronic anal fissure after a 3-year follow-up. METHODS: Eighty patients with chronic anal fissure were treated by whether open lateral internal sphincterotomy (group 1) or chemical sphincterotomy with 25 U botulinum toxin injected into the internal sphincter (group 2). Clinical and manometric results were analyzed. RESULTS: Overall healing was 92.5% in the open sphincterotomy group and 45% in the toxin botulinum group (P<.001). There is a group of patients with clinical (duration of disease >12 months and presence of a sentinel pile before treatment) and manometric factors (persistently elevated mean resting pressure, % of time presence of slow waves, and number of patients or the time presence ultra slow waves after treatment) associated with a higher recurrence of anal fissure. The final percentage of incontinence was 5% in the open sphincterotomy group and 0% in the botulinum toxin group (P>.05). CONCLUSION: We recommend surgical sphincterotomy as the first therapeutic approach in patients with clinical and manometric factors of recurrence. We prefer the use of botulinum toxin in patients older than 50 years or with risk factors for incontinence, despite the higher rate of recurrence, since it avoids the greater risk of incontinence in the surgical group. PMID- 15820456 TI - Gender differences among spouses of surgeons. AB - BACKGROUND: The goal of this study was to identify whether gender differences exist regarding the degree of spousal support for, satisfaction with, and perception of surgeons' careers. METHODS: A survey was performed of spouses of academic surgeons in the United States and Canada. RESULTS: Female respondents were significantly less likely to work outside of the home and were significantly more likely to be the major decision-makers at home. They were less likely to credit their surgeon spouses with contributing to household duties and childcare. If both spouses worked outside of the home, the female spouse was still more likely to be the major decision maker. Overall, male and female respondents rated their role in and their satisfaction with their spouses' career choices similarly. CONCLUSIONS: Male and female spouses contribute equally to the career choices of their surgeon spouses. However, female spouses, both surgeon and nonsurgeon, are more likely to make the majority of the decisions at home and contributions to household care. PMID- 15820457 TI - Relationship between secretory leukocyte protease inhibitor levels in bronchoalveolar lavage fluid and postoperative pulmonary complications in patients with esophageal cancer. AB - BACKGROUND: We investigated whether secretory leukocyte protease inhibitor (SLPI) is associated with pulmonary complications after esophagectomy. METHODS: We measured serial changes in the SLPI concentration in the bronchoalveolar lavage fluid (BALF) of 34 patients who underwent and examined the relationship between SLPI and postoperative morbidity. RESULTS: Fifteen (44%) of 34 patients (high group) had a BALF SLPI concentration >90,000 pg/mL at the end of the surgery (postoperative day [POD] 0). There was no significant difference between the high group and other 19 patients (low group) with respect to age, sex, preoperative comorbid conditions, tumor stage, surgical technique, operating time, or blood loss volume. Days of intubation and pulmonary complication rate were significantly increased in the high group compared with the low group. Logistic regression analysis revealed that the BALF SLPI level on POD 0 was significant for pulmonary complications. CONCLUSIONS: Our results indicate that assaying SLPI levels in BALF can be useful for the prediction of pulmonary complications after esophagectomy. PMID- 15820458 TI - Wide local excision or abdominoperineal resection as the initial treatment for anorectal melanoma? AB - BACKGROUND: Anorectal melanoma (AM) is a rare tumor with a poor prognosis. Treatment with abdominoperineal resection (APR) over wide local excision (WLE) is still debated. This study aimed to compare median survival of WLE and APR in patients with AM. METHODS: A systematic review of the literature was performed. Only series that allowed calculation of median survival were included. RESULTS: Fourteen studies met inclusion criteria. Average median survival of stage I WLE patients (N=34) and stage I APR patients (N=31) was 44 and 22 months, respectively (P=.001). For stage II patients, 7 underwent WLE, and 10 underwent APR with an average median survival of 36 and 14 months, respectively (P=.19). CONCLUSIONS: This study identified no stage-specific survival advantage to APR in favor of AM. Given that WLE is a more limited intervention associated with at least comparable survival, we propose that it be considered the initial treatment of choice for AM. PMID- 15820459 TI - Retained abdominal gallstones during laparoscopic cholecystectomy. AB - BACKGROUND: Although gallbladder perforation with spillage of bile and gallstones is quite common, the approach to retained gallstones in the abdomen still is controversial. METHODS: Laparoscopic cholecystectomy (LC) was performed on 580 patients with gallstones. In 101 (17%) patients, gallbladder perforations occurred during surgery, and in 43 (7%) patients, stone(s) were spilled into the peritoneal cavity. In 24 (4%) patients, gallstone(s) were not cleared entirely from the peritoneal cavity. These patients were invited to return for physical examination and biochemical tests. To investigate the retained abdominal gallstone(s) computed tomography was performed. RESULTS: Twenty-two patients were investigated. After a median follow-up period of 121 months, retained abdominal gallstone(s) were shown in 2 patients by computed tomography. Biochemical tests were normal except in 1 patient with chronic hepatitis. All of the patients were happy with their surgical results. CONCLUSIONS: This study revealed no harm caused by retained abdominal gallstone(s) during LC after long-term follow-up evaluation. PMID- 15820460 TI - A randomized trial of teaching bioethics to surgical residents. AB - BACKGROUND: Bioethics education has been recommended as a formal component of surgical residency training. The best method for teaching bioethics to residents is unclear. We compared the effectiveness of a standardized patient (SP)-based seminar to a traditional seminar format for teaching bioethics to surgical residents. METHODS: We randomized 31 first- and second-year surgical residents to either a SP-based seminar or a traditional seminar on informed consent. Immediately after the seminars, we evaluated resident performance in patient encounters on informed consent by using an objective structured clinical examination. Their knowledge of informed consent was also evaluated by using a 20 question short-answer written examination immediately after the seminars and then 3 weeks later. RESULTS: Twenty-nine residents completed the study; two withdrew because of an emergency. The SP seminar group had lower SP interview scores on the 22 item checklist compared with the traditional seminar group (57% versus 66%; difference -9%; 95% confidence interval [CI], -17% to -1%, P=.03). The SP seminar group also had lower knowledge scores on the questionnaire immediately after the seminar (60% versus 73%; difference -13%; 95% CI, -21% to -4%, P=.003). The difference in knowledge scores persisted at 3 weeks (41% for the SP group, 59% for the traditional seminar group; difference -18%; 95% CI, -29% to -7%; P=.002). CONCLUSIONS: A traditional seminar was superior to an SP-based seminar for teaching informed consent to surgical residents. PMID- 15820461 TI - A brief private group practice rotation changes junior medical students' perception of the surgical lifestyle. AB - BACKGROUND: Surgical practice is often perceived by students as a stressful and demanding lifestyle in which personal and family issues take low priority. For students to receive a more balanced view of surgical practice, we instituted a private practice preceptorship during the last week of our junior surgery clerkship in 2001. We hypothesized that a 4-day preceptorship with surgeons in private practice would improve student perception of surgery as a valid career choice without compromising student educational performance. METHODS: From January to December 2002, 107 junior medical students were assigned to a brief preceptorship with volunteer private practice surgeons during the surgery clerkship (group 1), while 28 students did not participate in the preceptorship (group 2). We assessed student satisfaction via questionnaire and compared student grade performance between groups. RESULTS: One hundred two questionnaires (95%) were returned. Overall, the preceptor experience was rated positively (9.0+/-0.1) based on a 10-point Likert scale. All students commented on the educational or enjoyment value of the preceptorship, with 44% specifically stating that the lifestyle in private surgical practice appeared pleasant. Interestingly, 9% of students volunteered that the experience had swayed them to consider surgery as a career. Twenty-four percent of questionnaires contained negative comments, mainly concerning not enough "hands-on" participation. Grades for the clerkship did not decrease in group 1 when compared with group 2 or with historic controls (n=113). CONCLUSIONS: A 4-day private practice preceptorship at the end of the junior surgery clerkship favorably alters student perceptions of a surgical career without diminishing student grade performance. PMID- 15820462 TI - Hyperbaric oxygen as adjuvant therapy in the management of necrotizing fasciitis. AB - BACKGROUND: Necrotizing fasciitis (NF) is an uncommon but serious infection of fascia and skin associated with considerable morbidity and mortality. One modality proposed for improving the outcome of this condition is hyperbaric oxygen (HBO) therapy. This is a form of medical treatment that involves intermittent inhalation of 100% oxygen under pressures exceeding the atmosphere. The aim of this article is to review current practice and evidence for the use of HBO as adjunctive therapy in the management of NF. METHODS: A survey of published English literature through searches of Medline and PubMed was carried out using the following key words: "necrotizing fasciitis," "Fournier's gangrene," "necrotizing soft tissue infections," "hyperbaric oxygen therapy," "and hyperbaric oxygen chambers." RESULTS: The results of studies on the use of HBO therapy in NF are inconsistent. Some studies have demonstrated that HBO can improve patient survival and decrease the number of debridements required to achieve wound control, whereas others have failed to show any beneficial effect. CONCLUSIONS: Encouraging results have been achieved with the addition of HBO therapy to standard treatment regimes, thus justifying further research in this field. More robust evidence by way of a prospective randomized trial is necessary before widespread and routine use of HBO in the management of NF can be recommended. PMID- 15820463 TI - The story of hyperbaric oxygen continues. PMID- 15820464 TI - A cumulative analysis of an individual surgeon's early experience with elective open abdominal aortic aneurysm repair. AB - BACKGROUND: Several studies have reviewed the role of hospital and surgeon case volumes in determining early mortality after elective open abdominal aortic aneurysm (AAA) repair. Few, however, have analyzed this relationship at the individual surgeon level. The purpose of this study was to display the usefulness of a unique statistical tool as a form of an ongoing practice audit. METHODS: All patients who underwent an elective open AAA repair by an individual surgeon at a university-affiliated medical center over a 5-year period were analyzed. The cumulative sum failure method was used to analyze the results over time. Failure was defined as the presence of early mortality, myocardial infarction, or a complication resulting in another surgical procedure or prolonged hospitalization. A target failure rate of 10% was chosen, and 80% alert and 95% alarm boundary lines were established. RESULTS: One hundred thirty-eight patients underwent elective AAA repair by this surgeon over a 5-year period (1998-2003). There were 5 early mortalities (3.6%), 15 myocardial infarctions (10.9%), and 3 major morbidities (2.2%). These results were plotted on a cumulative sum curve as an example of an ongoing practice audit. CONCLUSIONS: The cumulative sum failure method provides a tool whereby a surgeon can prospectively audit his practice and recognize trends in performance before their recognition by standard statistical tools. PMID- 15820466 TI - Initial use of a novel instrument to measure professionalism in surgical residents. AB - BACKGROUND: No universally accepted method to measure professionalism exists. We developed an instrument to measure specific aspects of professionalism in surgical residents. METHODS: Professionalism was deconstructed into 15 domains. Behavioral descriptors were determined for extreme and selected intermediate anchors. It became evident that residents could "go too far" in some professional behaviors. Therefore, although a 7-point continuous ordinal scale forms the framework, a score of 7 does not necessarily indicate the ideal. This characteristic minimizes the problem of inflated ratings. RESULTS: The instrument was utilized by attending faculty to evaluate residents and also by residents as a self-evaluation. Calculated from ordinal values, mean (SD) of the ratings across domains for faculty evaluations of residents was 4.95 (0.38) while mean for self-evaluations was 4.95 (0.39). Reliability was high (coefficient alpha 0.85). CONCLUSIONS: This instrument provides a means to measure professional behaviors during surgical residency. Repeated use will be required to thoroughly establish validity and reliability. PMID- 15820465 TI - Laparoscopic and thoracoscopic approaches for the treatment of hepatocellular carcinoma. AB - BACKGROUND: Laparoscopic/thoracoscopic intervention has been recently developed for hepatocellular carcinoma (HCC). However, no precise evaluation regarding this type of surgery has been performed. PATIENTS AND METHODS: A total of 33 laparoscopic and thoracoscopic treatments were performed on HCC patients, laparoscopic or thoracoscopic liver resection (LTR) in 15 patients and laparoscopic or thoracoscopic thermal ablation (LTA) in 18 patients. RESULTS: Postoperative hospital stay was 12.7 days after resection and 14.8 days after ablation. The 1- and 3- year actuarial survival rate were 100% and 80%, respectively, after LTR and 95% and 75%, respectively, after LTA. The 1- and 3- year disease-free survival rates were 75% and 40%, respectively, after LTR, and 60% and 28%, respectively, after LTA. CONCLUSION: LTR has the advantages of the curability of open surgery and less invasiveness of laparoscopic surgery. LTA is useful for tumors that cannot be percutaneously detected by ultrasound. Laparoscopic/thoracoscopic intervention is therefore considered an alternative means of treatment for selected HCC patients. PMID- 15820467 TI - Early retropancreatic dissection during pancreaticoduodenectomy. AB - BACKGROUND: During pancreaticoduodenectomy, early neck division may be inadequate, particularly in cases of replaced or accessory right hepatic artery (RHA) or diffuse intraductal papillary and mucinous neoplasm (IPMN) or when invasion of superior mesenteric artery is suspected. METHODS: Modification of the technique of pancreaticoduodenectomy with dissection of the superior mesenteric vessels performed from behind the head of the pancreas before any pancreatic or digestive transection. The pancreas is divided later, after adequate mobilization of the specimen from the vessels, on either the neck or the body, according to underlying disease. RESULTS: We used this technique successfully in 20 patients with a replaced or accessory RHA, which was successfully preserved in 19; one patient had inadvertent division of a 2-mm diameter accessory RHA that was ligated with no postoperative complications. Additionally, we used this technique in 18 patients with IPMN-14 pancreaticoduodenectomies extended to the body and 4 total pancreatectomies-and in 3 patients with adenocarcinoma involving the porto mesenteric confluence and needing en-bloc vascular resection. CONCLUSIONS: Early division of retroperitoneal margin is a useful technical variant of pancreaticoduodenectomy, which can be recommended in selective indications to improve safety and radicality of the procedure. PMID- 15820468 TI - Medical student concerns and fears before their third-year surgical clerkship. AB - BACKGROUND: This study describes the personal concerns of third-year medical students beginning their surgical clerkship. To assess these needs, we studied the expectations, interests, and concerns of beginning surgical clerks. METHODS: Beginning surgical clerks during the 2002-2003 academic year were invited to respond to a written survey. Responses were categorized into domains of interest and domains of concern. Analysis controlled for prior surgical experience, interest in a surgical career, and academic term. RESULTS: The response rate was 83%. Students were most curious about surgical practice, technical skills, and issues of lifestyle/balance among surgeons. Students were most concerned about fatigue, long hours, potential mental abuse, and poor performance. Differences between groups are described. CONCLUSIONS: Beginning surgical clerks have questions and concerns about topics often not addressed in the curriculum or syllabus. This study suggests the nature of improvements that could assist the inexperienced student in creating a positive clerkship experience and guiding their choice of career. PMID- 15820469 TI - Validation of a flexible endoscopy simulator. AB - BACKGROUND: Virtual reality (VR) simulation is a rapidly proliferating adjunct of surgical training. Numerous devices have evolved as educational tools in a variety of fields. Whether these tools can be used for validation of physicians' skills has yet to be determined. The objective of this study was to determine whether the GI Mentor (Simbionix, Lod, Israel) flexible endoscopy simulator construct could distinguish experienced endoscopists from beginners. METHODS: Seventy-five surgical attendings, fellows, and residents were recruited for participation in the study. Two cohorts were used and these groups were selected from 2 separate scientific sessions. Participants completed a standardized questionnaire documenting their endoscopic training and experience. Physicians subsequently were designated as experienced or beginner after their endoscopic training and experience were evaluated. All participants completed 1 of 2 colonoscopic simulations. The GI Mentor objectively evaluated performance on the basis of programmed data points, including the time to reach the cecum, the percentage of mucosa visualized, the completed polypectomy rate, the percentage of time spent in clear view through the lumen, the percentage of time that the patient was in pain, and overall efficiency. RESULTS: In both simulations, experienced endoscopists were more efficient than beginners (.32%/s vs. .26%/s, P=.02; and .53%/s vs. .37%/s, P=.03) and achieved a greater polypectomy rate (78% vs. 43%, P=.03; and 87% vs. 48%, P=.01). Furthermore, experienced endoscopists visualized more of the colonic surface (86% vs. 82%, P=.02) and spent a greater proportion of the time in clear view of the lumen (55% vs. 47%, P=.05) than beginners completing the first simulation. In the second simulation, experienced participants reached the cecum more rapidly than beginners (175 vs. 262 s, P=.01). CONCLUSIONS: The G1 Mentor VR colonoscopy construct appears valid. Significant performance differences were shown between the experienced and beginner cohorts. The beginner participants in this study were all physicians with some degree of endoscopic experience. Therefore, the G1 Mentor distinguished endoscopists of varying experience and exposure. Further validation studies are needed to evaluate the breadth of programs inherent to this simulator and to determine whether it may be used in the future for qualification and certification purposes. PMID- 15820470 TI - Hepatic portal venous gas. AB - BACKGROUND: Hepatic portal venous gas is an unusual entity associated with a variety of abdominal catastrophes. There is usually a grave prognosis when hepatic portal venous gas is associated with ischemic bowel disease. We reported a 57-year-old man with hepatic portal venous gas associated with extensive infarction of the jejunum and a concomitant perforation at a site in the terminal ileum leading to two operations performed 24 hours apart. Progressive ischemia and infarction after the initial laparotomy resulted in massive resection of the small bowel. METHODS: A follow-up abdominal radiography showed progressive dilatation of the small intestine and thickening of the bowel wall. Computed tomography of the abdomen showed pneumatosis intestinalis and gas collection within the intrahepatic and extrahepatic portal vein and superior mesenteric vein and free gas in the peritoneal cavity. RESULTS: At surgery, a long segment of ischemic change of the jejunum with focal necrosis and one perforation in the terminal ileum with no relation to the ischemic area was discovered. These two areas were resected respectively, and anastomosis was performed. Extensive necrosis of the residual bowel was found on the second-look operation performed 24 hours later, and subsequent resection of involved bowel was performed, resulting in a short-bowel condition. CONCLUSIONS: When hepatic portal venous gas associated with ischemic bowel disease is encountered, coexistence of other abdominal condition with no relation to ischemic segment should be considered. PMID- 15820471 TI - Letter to the editor. PMID- 15820472 TI - Standardization in laboratory medicine: new challenges. AB - The primary goal of Laboratory Medicine is to provide information that is useful to assist medical decision-making and permits optimal health care. This type of information should be independently obtained of the measurement test kits and instruments, and also of the laboratory where the procedure is carried out. It is therefore important to achieve a level of comparability of laboratory results among the many measurement procedures available so that results are harmonized and interchangeable over space and time. The standardization of measurements is therefore of high priority. In recent years, numerous efforts have been made at the international level under the auspices of the IFCC and other organizations to standardize measurement results for many important analytes, e.g. enzymes, cardiac proteins, etc. The aim of this review is to discuss some concepts related to the achievement of standardization by the implementation of a metrologically correct measurement system, providing some examples on how these concepts can be applied in Laboratory Medicine. PMID- 15820473 TI - A data base for 3000 monoclonal immunoglobulin cases and a new classification. AB - Data from monoclonal immunoglobulin cases screened in our laboratory for 40 years were used to assemble a cohort of 3094 cases selected according to immunoelectrophoresis and immunofixation interpretation and clinical data availability. Molecular distribution and original classification were used to establish five categories of cases: multiple myeloma (MM) and Waldenstrom macroglobulinemia (WM), other lymphoproliferative diseases, cases associated with heavy immunological diseases or other tumors, and benign cases. Free light chain (FLC) cases comprise 7% of the cohort, pluriclonal Ig, 6%. More than 50% of 1335 cases with identified diagnoses are not malignant hemopathies. Male/female ratio is less than 1 for MM cases and close to 1 for benign and other lymphoproliferative cases. Males are a majority in MW and associated cases. Follow-up periods range from 5 to 30 years for 263 cases. The main characteristics of this data base have been defined and the benefit of the original classification is highlighted. Future studies will investigate improvements resulting from immunofixation and current urinary analysis practices, as well as long-term follow-up, pluriclonal Ig cases, cryoglobulins, the meaning of the presence of FLC and malignant transformation. PMID- 15820474 TI - A minimally invasive technique for the evaluation of the regulatory steps of the two major pathways of bile acid synthesis. AB - BACKGROUND: Bile acid synthesis accounts for more than 95% of total cholesterol catabolism per day. We have developed a minimally invasive technique in humans that quantifies the rates of plasma appearance of 7alpha- and 27 hydroxycholesterol, representing the first steps of the "classical" and "alternative" pathways of bile acid synthesis, respectively. METHODS: For this purpose, during the intravenous infusion of synthetic deuterated isotopomers of 7alpha-hydroxycholesterol and 27-hydroxycholesterol plasma samples are collected and analysed by a GC-MS based method that allows to quantify the exogenous/natural isotopomer ratio of the two sterols. From this data, the rates of plasma appearance of 7alpha- and 27-hydroxycholesterol are calculated. RESULTS: In a group of healthy individuals steady state kinetics are obtained during a 2 h period yielding mean values of 2.0+/-0.8 and 3.7+/-0.6 mg/h for 7alpha- and 27-hydroxycholesterol, respectively. The data are consistent with findings using older techniques that require studies over several days. CONCLUSION: Considering that at steady state of the exogenous/natural isotopomer ratio the plasma appearance of the two regulatory hydroxysterols are related to the rate of bile acid synthesis via the "classical" and the "alternative" pathways, respectively, the proposed method could be used to evaluate the immediate effects of different diets and drugs and other determinants on cholesterol catabolism. PMID- 15820475 TI - Association of coronary artery ectasia with plasma insulin levels in Japanese men of heterozygous familial hypercholesterolemia with the low-density lipoprotein receptor gene mutation K790X. AB - BACKGROUND: Hyperinsulinemia is widely believed to be an important coronary risk factor. We investigated the effect of plasma insulin levels on the development of coronary ectasia in Japanese men with heterozygous familial hypercholesterolemia (FH). METHODS: A cross-sectional study was conducted in 20 FH men with the LDL receptor mutation (K790X) [age 42.3+/-2.8 years old, body mass index (BMI) 24.6+/ 0.7 kg/m2, total cholesterol (TC) 8.68+/-0.36 mmol/l, triglycerides (TG) 1.76+/ 0.23 mmol/l, high-density lipoprotein cholesterol (HDL-C) 0.977+/-0.065 mmol/l]. Subjects with diabetes mellitus were excluded. Plasma insulin levels, either fasting or during oral glucose tolerance test, were compared between subjects with and without coronary artery ectasia. RESULTS: FH subjects with coronary ectasia had significantly higher fasting plasma insulin levels than those without (12.6+/-1.4 vs. 7.7+/-0.5 mU/l; p<0.05). Also, plasma insulin levels during oral glucose tolerance test tended to be higher in the former than in the latter. CONCLUSIONS: Plasma insulin level could be an important determinant of the development of coronary artery ectasia in Japanese heterozygous FH men. PMID- 15820476 TI - Evaluation of two chemiluminescent immunoassays of ADVIA Centaur for hepatitis B serology markers. AB - BACKGROUND: This study evaluated the performance of 2 Bayer Hepatitis B Chemiluminescent Assays by comparing their performance to the corresponding DPC Immulite 2000 assays. METHODS: 953 samples were tested by both Immulite 2000 and Centaur HBsAg methods; and 75 samples were tested by both Immulite 2000 and Centaur anti-HBc IgM methods. RESULTS: The overall agreement between Immulite 2000 and ADVIA Centaur anti-HBc IgM assays was 100%. The agreement between Immulite 2000 and Centaur HBsAg assays was 100% for Immulite 2000 positive samples. The overall agreement for HBsAg negative samples was 99.9% after confirmation result was included. However, discrepancy between the 2 assays was observed in 53 samples which were tested initially as "weakly reactive"(index 1.0 1.95) but were not confirmed subsequently by the neutralization test on Immulite 2000. All 53 samples tested negative by Centaur HBsAg assay at the initial run. Furthermore, one Immulite 2000 HBsAg negative sample was reactive and confirmed by Centaur assay. CONCLUSIONS: While the performance of Bayer Hepatitis B HBsAg and aHBc IgM assays are in good agreement with corresponding DPC assays, the Bayer HBsAg assay is more efficient than the Immulite 2000 assay with its better separation of signal and background noise, and its complete automation features. PMID- 15820477 TI - Effect of caffeine on oxidation susceptibility of human plasma low density lipoproteins. AB - The effect of caffeine on oxidation susceptibility of low density lipoproteins (LDL) has been studied. LDL oxidation was induced by copper ions and an azo initiator. The conjugated dienes formation was followed spectrophotometrically and indicated the LDL oxidation status. Changes in LDL protein moiety during the lag phase, studied only in the experiments of copper induced oxidation, were followed using the intrinsic fluorescence spectroscopy. The decay of LDL fluorescence signal during initial stages of oxidation was slower in the presence of caffeine. Supported by the fluorescence quenching and polarization measurements, these results may indicate the protective role of caffeine against LDL oxidation in vitro. The results also indicate that the production of conjugated dienes in the propagation and decomposition phase of LDL oxidation is lower in the presence of caffeine, regardless of the initiation mechanism. These findings may have implications for the effect of caffeine on LDL in vivo. PMID- 15820478 TI - Thermodynamic study of beta-N-acetylhexosaminidase enzyme heterogeneity in human seminal plasma. AB - BACKGROUND: It has been suggested that the activity of beta-N acetylhexosaminidase (Hex) in seminal plasma may be used as a biochemical marker of azoospermia. The purpose of our study was to evaluate this hypothesis using a thermodynamic procedure developed to determine total Hex activity and that of its isoenzymes in this biological fluid. METHODS: Using the substrate 3,3' dichlorophenolsulphoftaleinil N-acetyl-beta-D-glucosaminide, a highly significant difference (p<0.001) is found between the activation energy of Hex A (41.5 kJ/mol) and of Hex B (72.3 kJ/mol), making it possible to determine the activity of these isoenzymes from the apparent activation energy of the total Hex in seminal plasma. RESULTS: A significant difference between the normozoospermic and azoospermic groups was only found for Hex A isoenzyme activity (p<0.05), although with considerable overlapping between the values of both groups. Significant partial correlations were found for the total Hex, Hex A and Hex B activities with the immobile spermatozoa count (p<0.01) and for total Hex and Hex B with the dead spermatozoa count (p<0.05). In turn, Hex A had a significant partial correlation with the live spermatozoa count (p<0.05); however, Hex activity in seminal plasma of acromosomal origin appears to be of little importance in quantitative terms. CONCLUSIONS: It was not possible to confirm that total Hex activity in seminal plasma, or even of its isoenzymes Hex A and Hex B, is a suitable biochemical marker of azoospermia (efficiency< or =67%). The thermodynamic procedure described may be a useful alternative for the study of the Hex enzyme heterogeneity in spermatozoa. PMID- 15820479 TI - Oxidative stress and antioxidant status in patients with alcoholic liver disease. AB - BACKGROUND: [corrected] Alcoholic liver diseases (ALD) are very common in lower socio-economical strata due to heavy drinking habits and multiple nutritional deficiencies. Ethanol causes liver damage by many mechanisms. The generation of lipid peroxidation by free radicals has been proposed as a mechanism for ethanol induced hepatotoxicity. These free radicals are destroyed by anti-oxidants. Many anti-oxidants are present in the diet, e.g., vitamin E, vitamin C etc. However, poor nutrition or malabsorption leads to deficiency of these vitamins. This may impair the anti-oxidative defense leading to ethanol induced oxidative stress and then to liver damage. METHODS: Oxidative stress and antioxidant defense were assessed in patients with alcoholic liver disease. Serum malondialdehyde (MDA) concentrations were measured as an index of lipid peroxidation, i.e., oxidative stress; and serum vitamins E and C concentrations were measured as an index of antioxidant status. RESULTS: Serum MDA concentrations were increased with the increase in severity of the disease. Concentrations of serum vitamins E and C were decreased in patients with alcoholic liver disease as compared to controls. CONCLUSIONS: Our observations may be due to increased demands of the same or increased utilization. PMID- 15820480 TI - Changes in immunomodulatory properties of Echinacea spp. root infusions and tinctures stored at 4 degrees C for four days. AB - BACKGROUND: Phytomedicinal preparations from members of the genus Echinacea are popular worldwide and frequently used to treat upper respiratory infections. With the increasing popularity of herbal medicines, many people are making their own Echinacea extracts at home and storing them at refrigerator (4 degrees C) temperatures. We tested the hypothesis that Echinacea extracts made using homemade methods change in immunomodulatory efficacy with storage at 4 degrees C over a 4-day period. METHODS: Three extract types (50% ethanol tincture, cold water infusion, hot water infusion) from 5 different species (Echinacea angustifolia, E. pallida, E. purpurea, E. sanguinea, E. tennesseensis) were prepared. Four in vitro immune assays (monocyte secretion of TNF-alpha, IL-10, and IL-12; and peripheral blood mononuclear cell proliferation) using human blood were used to test extract efficacy at Days 1 and 4 post-extraction. Two statistical analyses, traditional ANOVA and several statistical models that account for endotoxin effects, were used. RESULTS: Endotoxin was found to significantly impact immune outcomes only in 4-day old cold water infusions and not in all assays. Extracts showed the greatest stimulation in TNF-alpha assays. By extract type, 50% ethanol tinctures produced the most immune stimulation. By species, extracts from E. angustifolia extracts were the most efficacious in our assays; extracts from E. sanguinea showed the least activity overall. CONCLUSIONS: Taken together, these results suggest that: (1) homemade Echinacea extracts are efficacious in modulating immune cell activity in vitro but that their properties change with time during storage at 4 degrees C; and (2) endotoxin effects from extracts may be important considerations in the analysis of immunobiological data. PMID- 15820481 TI - Soluble transferrin receptor concentration is not superior to log ferritin for evaluating erythropoiesis in adolescents with iron deficiency anemia. AB - BACKGROUND: This study investigated the associations of soluble transferrin receptor (sTfR), log ferritin, the ratio of sTfR to log ferritin (sTfR-F index), and the log of the ratio of sTfR to ferritin [log(sTfR/F)] vs. reticulocyte production during iron deficiency. METHODS: Fluorescent intensity of reticulocytes, immature reticulocyte fraction (IRF), reticulocyte maturity index (RMI), sTfR, and serum ferritin were measured in 149 adolescents. RESULTS: There were no significant differences in reticulocyte parameters between the iron deficiency anemia (IDA) subjects with sTfR> or =4.9 mg/l and those with sTfR<4.9 mg/l. However, IDA subjects with log ferritin <0.73 microg/l exhibited significantly higher mean values for IRF and RMI, compared to those with log ferritin > or =0.73 microg/l (2.75+/-1.36% vs. 1.45+/-1.01%, p<0.05; 2.76+/-1.31% vs. 1.46+/-1.09%, p<0.05). In the non-IDA group, reticulocytes averaged 0.97+/ 0.31% in subjects with sTfR> or =2.1 mg/l, which were significantly above the values in those with sTfR<2.1 mg/l (0.72+/-0.16%, p=0.005), but no significant differences were observed in reticulocyte parameters between the subjects with log ferritin > or =1.35 microg/l and those with log ferritin <1.35 microg/l. Correlation coefficients of log ferritin vs. RMI (r=-0.41) were higher than those of sTfR, sTfR-F index, and log(sTfR/F) vs. RMI (r=0.24, r=0.30, and r=0.28, respectively) in IDA subjects. CONCLUSION: Reticulocytopoiesis is more closely associated with log ferritin value than with sTfR concentrations in IDA patients, although sTfR significantly reflects erythropoietic activity in non-IDA subjects. PMID- 15820482 TI - Reduction of urinary thiols in nephrotic syndrome--a possible effect of free iron. AB - BACKGROUND: Albumin is a potent antioxidant as it chelates transitional metals and contains antioxidants like thiol and bilirubin. In neprotic syndrome, the defining parameter is proteinuria with hypoalbuminemia. Therefore albuminuria in nephrotic syndrome may increase toxic transitional metal ions and also can cause loss of albumin associated antioxidants causing oxidative stress to the individual. METHODS: We investigated this possibility and estimated some markers of oxidative stress in 20 nephrotic syndrome patients and healthy controls along with urinary thiols, urinary bilirubin and plasma free iron in both cases and in the controls. RESULT: We found oxidative stress in 20 nephrotic syndrome patients and the markers of oxidative stress correlated significantly with proteinuria, but the urine of nephrotic syndrome patients (28.33+/-4.2 micromol/g creatinine)contained significantly less thiols compared to the healthy controls (88.45+/-10.6 micromol/g creatinine) and no biliribin. The patients plasma also showed free iron (0.7+/-0.05 micromol/l), a parameter undetectable in the healthy controls. CONCLUSION: We suggest that oxidative stress and presence of free iron in the patients were responsible for less thioluria and no bilirubinuria. A detailed study of oxidative biology in a large cohort of nephrotic syndrome patients is necessary to confirm the presence of free iron as appropriate chelation of free iron may benefit the long-term prognosis of the disease. PMID- 15820483 TI - Modulation of blood pressure, lipid profiles and redox status in hypertensive patients taking different edible oils. AB - BACKGROUND: Free oxygen radicals and insufficiency of antioxidants have been implicated in the pathogenesis of hypertension. We determined the effect of edible oils on blood pressure, lipid profiles and redox status in hypertensive patients given antihypertensive therapy (nifedipine-calcium channel blocker). METHODS: 530 patients medicated with nifedipine were divided into 3 groups (356 patients-sesame oil; 87 patients-sunflower oil; 47 patients-groundnut oil) and the control group (n=40) received only the drug, nifedipine. The respective oils were supplied to the patients and instructed to use as the only edible oil for 60 days, which comes to 35 g of oil/day/person. Blood pressure, lipid profiles [total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and triglycerides (TG)], lipid peroxidation [thiobarbituric acid reactive substances (TBARS)], enzymatic [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)] and nonenzymatic [(vitamin C, vitamin E, beta-carotene and reduced glutathione (GSH)] in blood were measured at baseline and after 60 days of oil substitution. RESULTS: Patients with nifedipine alone or with respective oils had significantly lowered blood pressure. TC, LDL-C and TG decreased while HDL-C elevated in sesame and sunflower oil groups. Increases of HDL-C and TG were noted in groundnut oil group. TBARS levels reduced in all the groups whereas the reduction was remarkable in sesame oil group. Activities of SOD elevated in the 3 oil groups whereas GPx and CAT increased only in sesame oil group. Levels of vitamin C, vitamin E, beta-carotene and GSH increased in sesame oil group whereas vitamin E and beta-carotene were elevated only in sunflower and groundnut oil groups. GSH increased in drug control group also. CONCLUSION: Among the 3 oils, sesame oil offers better protection over blood pressure, lipid profiles and lipid peroxidation and increases enzymatic and nonenzymatic antioxidants. PMID- 15820484 TI - Glutathione metabolism and antioxidant enzymes in patients affected by nonalcoholic steatohepatitis. AB - BACKGROUND: Oxidative stress and accumulation of excessive fat in the liver may underlie the pathophysiology of nonalcoholic steatohepatitis (NASH). Given that glutathione blood metabolism may represent an indicator of tissue oxidative status, we analysed the blood profile of various forms of glutathione in children with NASH, and we evaluated the presence of systemic oxidative stress by calculating the oxidised/reduced glutathione ratio (GSSG/GSH). Furthermore, we analysed the catalytic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione transferase (GST), and glutathione reductase (GR) in blood of patients. METHODS: Blood samples were obtained from 21 children with NASH and 28 controls. Total, reduced, oxidised, and protein-bound glutathione concentrations were determined by reversed-phase liquid chromatography with fluorescence detection. Antioxidant enzymes were spectrophotometrically assayed by using specific substrates. RESULTS: Our findings showed a 1.5-fold increase of GSSG in patients, resulting in a significant rise of the GSSG/GSH ratio. SOD, GPx, and GR activities were not significantly different in NASH respect to controls, whereas GST, which provides the second defence line against oxidative stress, was 17.8% increased. CONCLUSIONS: Our data demonstrate an impairment of glutathione metabolism and antioxidant enzyme activities in blood of patients with NASH, supporting a consistent role of free radical cytotoxicity in the pathophysiology of the disease. PMID- 15820485 TI - Glucose-6-phosphate dehydrogenase activity in term and near-term, male African American neonates. AB - BACKGROUND: We determined values for glucose-6-phosphate dehydrogenase (G-6-PD) activity in African American neonates. METHODS: G-6-PD activity was measured on umbilical cord blood from term and near-term healthy, male neonates. Neonates were stratified according to the number of neonates for each numerical unit of G 6-PD activity. Corrected end tidal carbon monoxide (ETCOc), a non-invasive index of hemolysis, was performed on each neonate. At least one predischarge transcutaneous bilirubin determination was performed. RESULTS: Five hundred neonates were studied. Two subpopulations were apparent, with no overlap between the subgroups. Mean value for the 64 (12.8%) infants with the lower values (G-6 PD deficient) was 2.7+/-1.1 U/g Hb, range 0.4-6.6 U/g Hb, while that for the 436 neonates with the higher values (G-6-PD normal) was 21.8+/-2.9 U/g Hb, range 14.5 33.8 U/g Hb. No significant differences in activity were noted between those neonates <37 weeks gestational age and those >37 weeks. Enzyme activity in the lower range in both groups was not related to the development of hyperbilirubinemia. G-6-PD enzyme activity did not correlate with ETCOc values either for the entire cohort or for the individual subsets. CONCLUSIONS: G-6-PD deficient neonates formed a separate subgroup from those with normal enzyme activity. The data supplied should facilitate interpretation of G-6-PD test results. PMID- 15820486 TI - Atheroprotective effect of exogenous heparin-derivative treatment on the aortic disturbances and lipoprotein oxidation in hypercholesterolemic diet fed rats. AB - BACKGROUND: The present work explores the myriad of biochemical and cellular changes that are featured in the early stages of atherosclerosis; if unchecked these changes lead to the complicated atherosclerotic plaque formation. The influence of a low-molecular-weight heparin derivative on the aortic aberrations and lipoprotein oxidation has been assessed in an experimental model of hypercholesterolemic atherogenesis. METHODS: Two groups of male Wistar rats (140+/-10 g) were fed a hypercholesterolemic atherogenic diet (rat chow supplemented with 4% cholesterol, 1% cholic acid and 0.5% thiouracil; CCT diet) for 2 weeks; one of these groups received LMWH (Certoparin) treatment of 300 microg/day/rat, s.c. for 7 days. An untreated control and a LMWH drug control group were also included. RESULTS: Abnormal increase in the aortic lipids glycosaminoglycans levels (p<0.001) in CCT-diet fed group was circumvented by the exogenous glycosaminoglycan (LMWH) treatment (p<0.001). The escalation of oxidative stress (markers-lipid peroxidation and thiol levels, superoxide dismutase and catalase activities) in the atherogenic aorta was minimised by LMWH treatment. Further, an increased susceptibility of the apo B-containing lipoproteins (LDL+VLDL) to oxidation in vitro, induced by copper ions and Fenton's reagent, was observed in the untreated CCT diet fed group. This paper reports the favorable modulation of these oxidative changes by LMWH administration. Vascular protection by LMWH is further substantiated by the normal aortic histologic findings as against the appearance of foam cells in the untreated atherogenic group. CONCLUSION: The exogenous heparin-derivative (LMWH) treatment attempted in this experimental model of hypercholesterolemic atherogenesis affords substantial protection against abnormal levels of aortic lipids and glycosaminoglycans, aortic oxidative stress and also stunts the lipoprotein peroxidative process, thereby proving its multi-faceted anti atherogenic effects. PMID- 15820487 TI - Valsartan reduces interleukin-1beta secretion by peripheral blood mononuclear cells in patients with essential hypertension. AB - BACKGROUND: Chronic low-grade inflammation response may contribute to the pathology of essential hypertension. Angiotensin II (Ang II) may be partly responsible for this process. Our early studies showed that individuals with essential hypertension had increased interleukin-1beta (IL-1beta) secretion by peripheral blood mononuclear cells (PBMCs). In this study, we investigated whether treatment with valsartan, an angiotensin receptor blocker, lowered IL 1beta secretion by PBMCs in patients with essential hypertension. METHODS: Twenty four patients with essential hypertension were randomized to treatment with valsartan (80 mg/day, group B) or matching routine therapy group (group A) for 2 weeks. PBMCs were isolated by gradient centrifugation. IL-1beta concentrations in supernatant from PBMCs were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with routine therapy group, patients treated with valsartan had decreased secretion of IL-1beta in PBMCs after stimulated by lipopolysaccharide (2857+/-643 vs. 2146+/-508 pg/ml, P<0.05). CONCLUSIONS: We suggest a direct anti-inflammatory effect of valsartan and a pro-inflammatory effect of Ang II in patients with essential hypertension. PMID- 15820488 TI - Anti-cyclic citrullinated peptide autoantibodies measured by an automated enzyme immunoassay: analytical performance and clinical correlations. AB - BACKGROUND: Autoantibodies against cyclic citrullinated peptide (anti-CCP) are considered to be a sensitive and specific marker for rheumatoid arthritis (RA). This study evaluated the analytical performance and clinical correlation of an automated enzyme immunoassay (DSX, DINEX Technologies), for the detection of anti CCP autoantibodies (DIASTAT anti-CCP, Axis-Shield, DUNDEE UK). METHODS: Commercial controls and serum pools were used to determine its precision, analytical sensitivity, functional sensitivity and linearity. Sera from 83 patients with established RA and from 140 controls, including patients with various autoimmune diseases, viral infections and cancer, as well as sex- and age matched healthy subjects, were studied. The rheumatoid factor (RF) was also assayed in each sample, and the results were compared to the anti-CCP findings. RESULTS: The total imprecision (CV%) was 4.7-7.2% for concentrations ranging between 1.98 and 71.81 U/mL. The lower detection limit was 0.038 U/mL. At a cut off of 5 U/mL, the sensitivity and specificity for RA were 67.5% and 99.3%, respectively. The RF had a sensitivity of 66.3% and a lower specificity 82.1% than anti-CCP. When the two antibodies were used together, the specificity was 99.1%. CONCLUSIONS: The anti-CCP assay we examined on a fully automated system showed a good analytical performance (analytical and functional sensitivity, linearity) and good clinical correlation. We conclude that this system can provide rapid, useful data. PMID- 15820489 TI - Identification of ethylsuccinylcarnitine present in some human urines. AB - Ethylsuccinylcarnitine, a previously undescribed acylcarnitine, was identified in urines obtained from 81% of adult volunteers. Its chemical structure was obtained by 1H nuclear magnetic resonance spectroscopy of the urinary purified compound and confirmed by its chemical synthesis. Its urinary excretion followed a circadian rhythm with a maximum occurring between 8 p.m. and 1 a.m. Excretion of this compound was enhanced after a load of L-carnitine and in this case, total 24 h urinary excretion may raise up to about 75 microM. It was observed that the day after adryamicine treatment, the compound was no more excreted. This molecule was absent or in trace amounts in urines obtained from few adults as well as in urines obtained from young subjects. In the positive urines, we detected an unknown organic acid whose excretion was almost parallel to that of ethylsuccinylcarnitine. PMID- 15820490 TI - Comparison of different cyclosporine immunoassays to monitor C0 and C2 blood levels from kidney transplant recipients: not simply overestimation. AB - BACKGROUND: Immunoassays used for the measurement of cyclosporine (CsA) usually show cross-reactivity for CsA metabolites, usually resulting in unacceptable bias. METHODS: To assess the performance of different immunoassays, CsA concentrations were analyzed in 132 samples using ACMIA, EMIT-VIVA, CEDIA-PLUS, and HPLC. Samples were collected from kidney transplant patients monitored with the traditional blood CsA trough level (C0, n=73) and the new sampling at 2-h post CsA dosing (C2, n=59). RESULTS: Overall, the correlations between HPLC and other methods were good (r values ranging from 0.85 to 0.97). The use of C2 concentrations to monitor CsA exposure were associated with an overall better performance of all the immunoassays as compared with C0 values. However, none of the immunoassays agreed with the guidelines proposed in the Lake Louis Consensus Conference. Of note, the CEDIA-PLUS was the only that provided a linear relationship with HPLC for both sampling times. A false positive case associated with ACMIA was also documented in blood samples from a patient withdrawn from CsA for 1 month. CONCLUSION: These data suggest that the performance of some of the most used immunoassays is not satisfactory, eventually leading to incorrect therapeutic decision guided by erroneous CsA monitoring. PMID- 15820491 TI - Prevalence of myocardial infarction is related to hyperhomocysteinemia but not influenced by C677T methylenetetrahydrofolate reductase and A2756G methionine synthase polymorphisms in diabetic and non-diabetic subjects. AB - BACKGROUND: Hyperhomocysteinemia has emerged as a novel risk factor for myocardial infarction (MI). Some mechanisms proposed to explain its relationship with coronary events are also shared by major coronary risk factors. We examined whether C677T methylenetetrahydrofolate reductase and A2756G methionine synthase polymorphisms could affect the relative risk for MI. METHODS: A sample of 196 individuals was divided into four groups (diabetics with MI, n=43; diabetics without MI, n=50; non-diabetics with MI, n=47; non-diabetics without MI, n=56) and compared regarding the prevalence of the polymorphisms, risk factors, and biochemical parameters. RESULTS: Higher prevalence of hyperhomocysteinemia was found in MI patients (p<0.05 vs. non-MI subjects), in males (p<0.001 vs. female) and in those > or = 65 years (p=0.01 vs. <65 years). Homocysteine was negatively associated with HDL-C (p<0.05) and glucose, although results did not reach significance (p=0.06). Similar distribution of studied polymorphisms was seen in all groups, which presented normal folate and vitamin B12 serum levels. CONCLUSIONS: Higher homocysteinemia was predominantly observed in men, presenting low HDL-C, and at advancing age. Methylenetetrahydrofolate reductase and methionine synthase polymorphisms did not contribute to risk assessment in diabetic and non-diabetic subjects presenting normal folate levels. PMID- 15820492 TI - Oxidative stress on mitochondrial antioxidant defense system in the aging process: role of DL-alpha-lipoic acid and L-carnitine. AB - BACKGROUND: Oxidative damage is hypothesized to accumulate throughout the lifetime of an organism, eventually giving rise to aging. The mitochondria may be the primary cellular source and target of endogenous ROS as they are produced as a normal byproduct of the electron transport system. METHODS: Male albino Wistar rats were used in this study. The animals were divided into 6 groups, each group consisting of 6 animals. Groups I, III, and V were young, middle-aged and aged control rats and Groups II, IV, and VI were treated with carnitine (300 mg/kg bw) and dl-alpha-lipoic acid (150 mg/kg bw), respectively. After the treatment period, the animals were sacrificed and the heart and skeletal muscle were removed for analysis. RESULT: There was a significant reduction in the levels of antioxidants in both middle-aged and aged rats whereas the thiobarbituric acid reactive substances were found to be increased. Co-supplementation of carnitine and lipoic acid improved the antioxidant status and brought down the levels of TBARS. CONCLUSION: Co-supplementation of lipoic acid with carnitine has a beneficial effect in reversing the age-related abnormalities seen in aging. This effect was associated with the decrease in free radical production and rise in antioxidant levels by carnitine and lipoic acid, thereby lowering oxidative stress. PMID- 15820493 TI - Pilot study of arbitrarily primed PCR-single stranded DNA conformation polymorphism analysis for screening genetic polymorphisms related to specific phenotypes. AB - BACKGROUND: To investigate relationships between phenotypes and genotypes is not simple. We propose a phenotype-to-genotype screening strategy and pooled DNA system. As a pilot study of this strategy, we used arbitrarily primed polymerase chain reaction (AP-PCR) in combination with single-stranded DNA conformation polymorphism (SSCP) to screen for genetic polymorphisms associated with longevity. METHODS: Study subjects were separated into 3 age groups, individuals aged >100 years, 90-99 years and 60-69 years. Genomic DNAs were prepared from each individual, pooled to represent the 5 study groups, and then the pooled genomic DNAs were subjected to AP-PCR-SSCP analysis. RESULTS: We found 1 SNP more frequently in senior citizens with longevity. The genotype frequency of the 82133G>A polymorphism of human chromosome 3 clone RP11-61K12 (AC011199) differed significantly (P=0.0189, Fisher's exact test) between older subjects (>90 years) and younger subjects (<70 years). It is noteworthy that the strategy we describe herein was useful for identifying an SNP that showed statistically significant differences in its distribution across the subject groups. CONCLUSIONS: The pooled DNA strategy and quantitative genotype discrimination can also be applied to screening for the relationship between phenotype and genotype more effectively. PMID- 15820494 TI - Rapid detection of the SPINK5 polymorphism Glu420Lys by real-time PCR technology. AB - BACKGROUND: The SPINK5 gene, encoding the serine protease inhibitor LEKTI maps to chromosome 5q32, and has been suggested to be a locus predisposing to atopic diseases in general. The Glu420Lys variant showed significant association with atopy, asthma and atopic dermatitis in recent studies. AIMS OF THE STUDY: Development of a high throughput assay to analyse the polymorphism G1258A (Glu420Lys) in exon 14 of the SPINK5 gene followed by the validation using samples of 235 latex-allergic health care workers (HCWs) with (N=63) and without asthma (N=172), and 80 non-atopic controls. METHODS: Twenty DNA samples were first analysed by a polymerase chain restriction fragment analysis (RFLP) using Hph I to generate defined control DNAs which were used for the development of the assay suitable for the detection of the Glu420Lys variant by LightCycler technology. RESULTS AND CONCLUSIONS: 315 samples were successfully screened with this new assay. The temperatures in the melting analysis of the SPINK5 exon 14 PCR product were characteristic to the probes hybridised to the mutant (AA) at 51.5 degrees C and to the wild-type (GG) at 59.5 degrees C. The fast and reliable mutation detection in the tested samples makes this high-speed method suitable for larger epidemiological studies. PMID- 15820495 TI - Increased levels of CD40-CD40 ligand system in patients with essential hypertension. AB - BACKGROUND: Although it has been hypothesized that hypertension is in part an inflammatory disorder, clinical data linking inflammation with incident hypertension are scarce. There is evidence that have shown that CD40-CD40L interaction plays a pathogenic role in inflammatory disorders. We assessed whether CD40 system expressions were altered in patients including 30 with hypertension grade 1, 80 with hypertension grade 2 and 40 with hypertension grade 3. METHODS: Twenty normal controls and 150 patients with essential hypertension were investigated. The expression of CD40 and CD40L on platelet was analyzed by indirect-immunofluorescence flow cytometry and soluble CD40L level was determined by a commercially available ELISA. C-reactive protein was also measured by ELISA. RESULTS: All patients with hypertension showed a significant increase of CD40 (67.1+/-9.6 Mean Fluorescence Intensity, MFI) and CD40L (15.3+/-5.0 MFI) coexpression on platelets as well as sCD40L levels (12.8+/-3.9 ng/ml ) compared with controls (p<0.0001). We found that CRP levels related to CD40-CD40L system. We also observed a slight correlation between sCD40L level and blood pressure. During 3 months follow-up, patients with enhanced levels of sCD40L (>15 ng/ml) indicated a tough control of blood pressure. CONCLUSION: Patients with essential hypertension show increased coexpression of CD40 system, which suggests that hypertension is in part an inflammatory disorder. PMID- 15820496 TI - Attenuation of serum lipid abnormalities and cardiac oxidative stress by eicosapentaenoate-lipoate (EPA-LA) derivative in experimental hypercholesterolemia. AB - BACKGROUND: Dietary cholesterol plays an important role in development of atherogenesis and cardiovascular disease. We explored the lipemic-oxidative injury in the hypercholesterolemic atherogenic animals. The effects of eicosapentaenoic acid (EPA), dl-alpha-lipoic acid (LA) and eicosapentaenoate lipoate derivative (EPA-LA) were tested for their efficacy in controlling the atherogenic disturbances. METHODS: Four groups of male Wistar rats were fed with a high cholesterol diet (rat chow supplemented with 4% cholesterol and 1% cholic acid; HCD) for 30 days. Of these groups, 3 groups of rats were treated with either EPA (oral gavage, 35 mg/kg body weight/day), LA (oral gavage, 20 mg/kg body weight/day) or EPA-LA derivative (oral gavage, 50 mg/kg body weight/day) from 16th day to 30th day of the experimental period. RESULTS: HCD induced abnormal increase in lipid peroxidation and serum cholesterol, triglyceride, LDL and VLDL, and a decreased HDL concentration. Altered activity of cardiac and serum creatine kinase, accompanied by a depressed cardiac enzymatic and nonenzymatic antioxidants defense system were observed in HCD fed rats. These changes were partially restored in the EPA and LA treated groups, however, their combined derivative EPA-LA more effectively restored the altered parameters near to that of control (P<0.05). CONCLUSION: Lipid abnormalities and oxidative injury were induced by a hypercholesterolemic diet. Administration of the combination treatment of EPA-LA afforded protection against the lipemic-oxidative injury. PMID- 15820497 TI - Pediatric reference intervals for FSH, LH, estradiol, T3, free T3, cortisol, and growth hormone on the DPC IMMULITE 1000. AB - BACKGROUND: We studied serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), triiodothyronine (T3), free T3 (FT3), cortisol and growth hormone (GH) concentrations in a population of pediatric patients. The reference intervals were determined separately for females and males stratified by age groups to assess age- and sex-related differences. Our objective was to obtain reference intervals for the 7 serum analytes for our pediatric population using the IMMULITE 1000 system. METHODS: Serum samples of 800 in- and out patients, newborn to 19 years old were analyzed using the DPC IMMULITE 1000 chemiluminescent immunoassay system. RESULTS AND CONCLUSIONS: We report pediatric reference intervals for FSH, LH, E2, T3, FT3, cortisol, and GH. These reference intervals provide the basis for clinical interpretation of laboratory results using the IMMULITE 1000 system and the assessment of child development. PMID- 15820498 TI - Inter-laboratory differences in sirolimus results from six sirolimus testing centers using HPLC tandem mass spectrometry (LC/MS/MS). PMID- 15820499 TI - Role of hs-CRP measurements in the current cardiovascular risk assessment. PMID- 15820500 TI - A serine proteinase inhibitor isolated from Tamarindus indica seeds and its effects on the release of human neutrophil elastase. AB - Proteinaceous inhibitors with high inhibitory activities against human neutrophil elastase (HNE) were found in seeds of the Tamarind tree (Tamarindus indica). A serine proteinase inhibitor denoted PG50 was purified using ammonium sulphate and acetone precipitation followed by Sephacryl S-300 and Sephadex G-50 gel filtration chromatographies. Inhibitor PG50 showed a Mr of 14.9 K on Sephadex G 50 calibrated column and a Mr of 11.6 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. PG50 had selective activity while cysteine proteinases (papain and bromelain) and serine proteinases (porcine pancreatic elastase and bovine chymotrypsin) were not inhibited, it was strongly effective against serine proteinases such as bovine trypsin and isolated human neutrophil elastase. The IC50 value was determined to be 55.96 microg.mL-1. PG50 showed neither cytotoxic nor haemolytic activity on human blood cells. After pre incubation of PG50 with cytochalasin B, the exocytosis of elastase was initiated using PAF and fMLP. PG50 exhibited different inhibition on elastase release by PAF, at 44.6% and on release by fMLP, at 28.4%. These results showed that PG50 preferentially affected elastase release by PAF stimuli and this may indicate selective inhibition on PAF receptors. PMID- 15820501 TI - Functional proteome of bones in rats with osteoporosis following ovariectomy. AB - Osteoporosis is a chronic condition chiefly affecting postmenopausal women, in whom the skeleton loses a significant percentage of its mineralized mass and mechanical resiliency, thereby becoming prone to fracture. Although the effect of the loss of estrogen on bone metabolism has been documented, its mechanism is still poorly understood. In the present proteomic study, we characterized the effect of estrogen deficiency on protein expression in rat bones. Using two dimensional gel electrophoresis, mass spectrometry and rat protein database, we successfully identified three distinctly changed proteins named thioredoxin peroxidase 1, myosin light polypeptide 2 and ubiquitin-conjugating enzyme E2-17 kD, among which ubiquitin-conjugating enzyme E2-17 kD has been documented to be an estrogen-related protein, but the other two are first reported to be osteoporosis-related proteins in the current study. These results provide valuable experimental evidences for the elucidation of the molecular mechanism of osteoporosis related to the loss of estrogen. PMID- 15820502 TI - Insulin stimulates Ca2+ uptake via PKC, cAMP, and p38 MAPK in mouse embryonic stem cells. AB - Embryonic stem (ES) cells are provided as a powerful tool for developmental biology and have been shown to respond to insulin. However, little is known about the effect of insulin on [Ca2+]i regulation in the ES cells, although many cellular functions are tightly regulated by [Ca2+]i. Therefore, we examined the effect of insulin on Ca2+ uptake and its related signal pathways in the mouse ES cells. Mouse ES cells expressed alkaline phosphatase (AP), transcription factor Oct-4, and stage-specific embryonic antigen-1 (SSEA-1). Insulin increased the Ca2+ uptake in a time- and dose-dependent manner and the effect was blocked by L type Ca2+ channel blockers, nifedifine and methoxyverapamil. Genistein or herbimycin A (tyrosine kinase inhibitors), wortmannin (PI-3K inhibitor), and staurosporine or bisindolylmaleimide I (PKC inhibitors) completely prevented insulin-induced increase of Ca2+ uptake. Wortmannin blocked insulin-induced PKC activation, but SQ 22536 (adenylate cyclase inhibitor) did not. Insulin also rapidly increased formation of inositol phosphates (IPs). We examined the involvement of MAPKs in mediating the effect of insulin on Ca2+ uptake. SB 203580 (p38 MAPK inhibitor) but not PD 98059 (p44/42 MAPKs inhibitor) blocked insulin induced increase of Ca2+ uptake. Insulin significantly increased the phosphorylation of p38 MAPK but not p44/42 MAPKs. In addition, genistein, PKI, and bisindolylmaleimide I blocked the phosphorylation of p38 MAPK by insulin, suggesting a causal relationship. In conclusion, insulin partially stimulated Ca2+ uptake via PKC, cAMP, and p38 MAPK signaling pathways in mouse ES cells. PMID- 15820503 TI - Different vanilloid agonists cause different patterns of calcium response in CHO cells heterologously expressing rat TRPV1. AB - The vanilloid receptor subtype 1 (TRPV1 or VR1) is expressed in nociceptive primary afferents of the C-fiber 'pain' pathway and has attracted considerable attention as a therapeutic target. Here, using rat TRPV1 heterologously expressed in Chinese hamster ovary cells, we show that different agonists show different patterns of modulation of the intracellular Ca2+ concentration, monitored in individual cells by fura-2 Ca2+ imaging. We identified 5 parameters (potency, maximal response, latency of response, variability of latency of response among individual cells, and desensitization) which behaved differently for different compounds. The potencies of the compounds examined ranged from EC50 values of 80 pM to 9 microM. Peak levels of induced [Ca2+]i were observed either higher (RTX) or lower (anandamide) than for capsaicin. Significant latencies of response were observed for some (e.g. RTX) but not other derivatives, with great variation among individual cells in this latency. Marked desensitization after stimulation was detected in some cases (e.g. anandamide, capsaicin); for others, no desensitization was observed. We conclude that structurally diverse vanilloid agonists induce marked diversity in the patterns of Ca2+ response. This diversity of response may provide opportunities for pharmacological exploitation. PMID- 15820504 TI - A novel L-glutamate transporter inhibitor reveals endogenous D-aspartate homeostasis in rat pheochromocytoma MPT1 cells. AB - We previously reported for the first time that D-aspartate (D-Asp) is biosynthesized by cultured mammalian cells such as pheochromocytoma (PC)12 cells and its subclone MPT1 (FEBS Lett. 434 (1998) 231, Arch. Biochem. Biophys. 404 (2002) 92). We speculated that D-Asp levels in the intra- and extracellular spaces of the cultured cells are maintained in a dynamic state of homeostasis. To test this here, we utilized a novel and potent L-Glu transporter inhibitor, TFB TBOA. This inhibitor proved to be a genuine nontransportable blocker of the transporter even during long periods of culture. Use of this inhibitor with MPT1 cells confirmed that D-Asp levels are in a dynamic steady state where it is constantly released into the extracellular space by a yet undefined mechanism as well as being constantly and intensively taken up by the cells via the L-Glu transporter. We estimated the rate with which D-Asp is constitutively released from MPT1 cells is approx. 3.8 pmol/h/1x10(5) cells. PMID- 15820505 TI - Functional analysis of cells obtained from bronchoalveolar lavage fluid (BALF) of lung cancer patients. AB - BALF from tumor segments provides access to immune system cells in contact with lung tumors. We analyzed BALF cells as to their production of H2O2 and NO, comparing tumor-affected to non-affected lung segments. Twelve patients were studied (4 NSCLC, 3 SCC, 5 Adenocarcinoma). The cell numbers recovered from BALF varied, and, in adenocarcinoma patients, smaller numbers were recovered from tumor-affected segments. H2O2 production (up to 6.3 nmoles/2x10(5)cells) was obtained in 7/12 patients and, in these, it was more frequent in non-affected segments (7/7) than in affected segments (2/7). After culture, NO production was observed in three patients (6 to 314 microM) that also produced H2O2. These functional characteristics of cells in contact with neoplasia may have a role in determining the fate of the interactions between the immune system and lung cancer. PMID- 15820506 TI - Decreased tyrosine hydroxylase activity in the adrenals of spontaneously hypertensive rats. AB - Higher activity of the peripheral sympathetic nervous system, accompanied by higher tyrosine hydroxylase activity is frequently and consistently reported in human essential hypertension as well as in animal models of hypertension. However, results obtained in the adrenals, particularly in young animals before the development of hypertension, are scarce and controversial. In the present study tyrosine hydroxylase activity and catecholamine content in the adrenals of spontaneously hypertensive rats and of age-matched control Wistar Kyoto rats were evaluated before, during and after the development of hypertension (5, 12 and 22 week-old animals). Results show that both tyrosine hydroxylase activity and total amine content in the adrenals of spontaneously hypertensive rats were significantly reduced (35% reduction) at all studied ages. Determination of the kinetic parameters for tyrosine hydroxylase in the adrenals of 5 week-old spontaneously hypertensive rats revealed a 38% reduction in V(max) values (13.4 versus 21.3 nmol L-DOPA/mg prot/h in age-matched controls) accompanied by lower levels of expression of both tyrosine hydroxylase total protein and phosphoSer40 observed by Western-Blot. In contrast, norepinephrine content in both plasma and tail artery were significantly higher in the spontaneously hypertensive strain. In conclusion, contrary to the higher peripheral sympathetic activity, tyrosine hydroxylase activity and catecholamine content in the adrenals of spontaneously hypertensive rats are markedly reduced before, during and after the development of hypertension. End product, long-term feedback inhibition by the high norepinephrine plasma levels could be responsible for this reduction, establishing yet another regulatory mechanism of tyrosine hydroxylase operating in adrenal cromaffin cells. PMID- 15820507 TI - Monochloramine induces acute and protracted colitis in the rat: response to pharmacological treatment. AB - Monochloramine is a powerful oxidative molecule that is produced in inflammatory sites. We investigated the effect of intrarectally administered monochloramine (3.2 mg) in the rat. A single enema induced after 24 h an intense inflammatory reaction characterized by mucosal necrosis, submucosal edema, hemorrhage and colonic thickening, as well as induction of nitric oxide synthase and tumor necrosis factor and an increase in the interferon gamma/interleukin 4 ratio. The inflammatory response peaked 3-5 days after monochloramine administration and then followed a extended recovery phase. At 1 week there was substantial but incomplete mucosal repair, submucosal edema, neutrophil/macrophage infiltration and increased myeloperoxydase and alkaline phosphatase activities. Oxidative stress, as determined by malonyldialdehyde levels, was prominent only in the acute phase (3-5 days). Monochloramine colitis was amenable to pharmacological treatment with sulphasalazine or prednisolone, suggesting that it may be used as an experimental model of inflammatory bowel disease. In conclusion, monochloramine induces acute and protracted colonic inflammation in the rat. Locally produced monochloramine might contribute to the perpetuation of inflammatory bowel disease. PMID- 15820508 TI - In vitro antioxidant activity of polyphenol extracts with antiviral properties from Geranium sanguineum L. AB - Recent evidence shows that plant polyphenols exhibit antioxidant and radical scavenging properties. By three separate and complementary methods--DPPH assay, beta-carotene-linoleic acid assay and NBT-reduction assay it was established that a polyphenol-rich extract from the medicinal plant Geranium sanguineum L. with strong anti-influenza virus activity, possessed antioxidant and radical scavenging capacities. For comparative reasons caffeic acid and the synthetic antioxidant BHT were used. Total soluble phenolic constituents of the MeOH extract measured by Folin-Ciocalteu reagent were found as 34.60% (w/w). Further it was demonstrated that the EtOAc fraction, retaining the majority of the in vivo protective effect exhibited a strong O2-scavenging activity while the n-BuOH fraction, containing the majority of the in vitro antiviral activity provoked generation of O2-. The O2- scavenging activity of all three preparations correlated with the rate of the protective effect shown in the murine model of experimental influenza virus infection. The present results are in accordance with our intensive studies on the mode of the protective effect of the plant extract which showed positively that the protection may possibly be attributed to the combination of more than one biological activities and that the use of antioxidants might be an useful approach in the treatment of influenza infection. PMID- 15820509 TI - Effects of black grape extract on activities of DNA turn-over enzymes in cancerous and non cancerous human colon tissues. AB - Effects of extract of dried whole black grape including seed on adenosine deaminase (ADA), 5' nucleotidase (5'NT) and xanthine oxidase (XO) enzymes were investigated in cancerous and non-cancerous human colon tissues. Enzyme activities were measured in 20 colon tissues, 10 from cancerous region and 10 from non cancerous region with and without pre incubation with black grape extract. ADA and 5'NT activities were found increased and that of the XO decreased in the cancerous tissues relative to non cancerous ones. After incubation period with black grape extract for 12 h, ADA and 5'NT activities were found to be significantly lowered but that of XO unchanged in both cancerous and non cancerous tissues. Results suggest that ADA and 5'NT activities increase but XO activity decreases in cancerous human colon tissues, which may provide advantage to the cancerous tissues in obtaining new nucleotides for rapid DNA synthesis through accelerated salvage pathway activity. Black grape extract makes significant inhibition on the ADA and 5'NT activities of cancerous and non cancerous colon tissues, thereby eliminating this advantage of cancer cells, which might be the basis for the beneficial effect of black grape in some kinds of human cancers. PMID- 15820510 TI - The influence of attending to multiple locations on eye movements. AB - The present paper reports results of a dual task study in which two locations were endogenously cued as possible target locations, while only one eye movement had to be executed. During the cue period, letters were briefly presented at the saccade goals and at no-saccade goals. Results show that performance was better for letters presented at any of the saccade goals than for letters presented at the no-saccade locations. Furthermore saccades deviated away from the non saccaded target location, suggesting inhibition of the location to which the eyes should not go. The results indicate that the premotor theory also holds for conditions in which attention is allocated to multiple locations. PMID- 15820511 TI - Associating color appearance with the cone chromaticity space. AB - A cone chromaticity space, a transform of a colorimetric specification system into coordinates that represent cone excitations, does not provide color appearance information. Boynton and Olson (Color Research and Application 12, 94 105, 1987) gathered color naming for the 424 Optical Society of America Uniform Color Scales (OSA-UCS) color samples. Here, a computational algorithm was developed that converts OSA-UCS sample values into L, M, S cone excitations based on the 1964 CIE 10 degrees Standard Observer. This makes it possible to plot the cone chromaticities associated with the eight color names used by Boynton and Olsen's observers to describe the non-dark appearing colors. PMID- 15820512 TI - Eccentric perception of biological motion is unscalably poor. AB - Accurately perceiving the activities of other people is a crucially important social skill of obvious survival value. Human vision is equipped with highly sensitive mechanisms for recognizing activities performed by others [Johansson, G. (1973). Visual perception of biological motion and a model for its analysis. Perception and Psychophysics, 14, 201; Johansson, G. (1976). Spatio-temporal differentiation and integration in visual motion perception: An experimental and theoretical analysis of calculus-like functions in visual data processing. Psychological Research, 38, 379]. One putative functional role of biological motion perception is to register the presence of biological events anywhere within the visual field, not just within central vision. To assess the salience of biological motion throughout the visual field, we compared the detectability performances of biological motion animations imaged in central vision and in peripheral vision. To compensate for the poorer spatial resolution within the periphery, we spatially magnified the motion tokens defining biological motion. Normal and scrambled biological motion sequences were embedded in motion noise and presented in two successively viewed intervals on each trial (2AFC). Subjects indicated which of the two intervals contained normal biological motion. A staircase procedure varied the number of noise dots to produce a criterion level of discrimination performance. For both foveal and peripheral viewing, performance increased but saturated with stimulus size. Foveal and peripheral performance could not be equated by any magnitude of size scaling. Moreover, the inversion effect--superiority of upright over inverted biological motion [Sumi, S. (1984). Upside-down presentation of the Johansson moving light-spot pattern. Perception, 13, 283]--was found only when animations were viewed within the central visual field. Evidently the neural resource responsible for biological motion perception are embodied within neural mechanisms focused on central vision. PMID- 15820513 TI - Structural details of rat extraocular muscles and three-dimensional reconstruction of the rat inferior rectus muscle and muscle-pulley interface. AB - Light microscopy, electron microscopy and morphometry revealed structural details and allowed generation of a three-dimensional reconstruction of the pulley and muscle-pulley interface of extraocular muscle. The inferior rectus orbital layer was bifurcate in shape and extended anterior to the pulley. The putative pulley structure itself was asymmetric; loosely attached at the orbital aspect it adhered tightly to the global aspect of muscle. Orbital multiply innervated fiber proportion increased anterior to the pulley insertion site. Additionally longitudinal variation in juxtaposition of orbital and global layers was noted. These newly described structural details provide novel mechanistic insight for extraocular muscle function in rats. PMID- 15820514 TI - Investigating the light absorption in a single pass through the photoreceptor layer by means of the lipofuscin fluorescence. AB - Reflection densitometry has been widely used to measure the density difference of the bleachable cone photopigments in human eyes. Most such measurements make a series of assumptions concerning the amount of scattered light to derive an estimate of the true cone photopigment density from the density difference measurements. The current study made three types of measurements of the light returning from the eye before and after bleaching: the amount of light returning in the "directed" reflection, which is a double-pass estimate of the cone photopigment density; the amount of light in undirected or diffuse reflection; and the amount of fluorescence from lipofuscin in the RPE, which provides a single-pass measurement of optical density difference. For a 1 deg foveally fixated field, the density difference estimates for the three measurements were 0.68, 0.21, and 0.22 respectively. The lipofuscin fluorescence was found to be unguided. The background density difference was non-negligible and very close to the single pass estimate from fluorescence. These measurements each involve potentially different pathways of light through the retina, and therefore place different constraints on models of these pathways. A simple model comparing the directional and the fluorescence optical densities produced retinal coverage estimates around 70-75%. Estimates of the shape factor of the single pass optical Stiles-Crawford effect were evaluated from the dark-adapted and bleached fluorescence measurements. The values were closer to those obtained from psychophysical methods than to the double pass optical Stiles-Crawford shape factors obtained directly from retinal reflectometry. PMID- 15820515 TI - Noticeable, troublesome and objectionable limits of blur. AB - We investigated limits at which induced blur becomes noticeable, troublesome and objectionable. We used 15 cyclopleged subjects, a Badal optometer with lines of three high contrast letters as targets, 3-6 mm artificial pupils, and 0.0-0.7 logMAR letter sizes. For 0.0 logMAR size, mean "noticeable" blur limits were +/ 0.33D, +/-0.30D and +/-0.28D at 3 mm, 4 mm and 6 mm, respectively, but increased by about 70% for 0.7 logMAR letters. All limits reduced by about 17% as pupil size increased from 3 mm to 6 mm. Letter size had a significant influence on all blur limits (1.6-2.1 times), but blur direction had no significant effect. Magnitudes of "troublesome" and 'objectionable" limits were 1.6-1.8 times and 2.1 2.5 times relative to "noticeable" limits, respectively. Our results suggest criteria for troublesome and objectionable blur are relatively unaffected by letter size. PMID- 15820516 TI - Integrating visual cues for motor control: a matter of time. AB - The visual system continuously integrates multiple sensory cues to help plan and control everyday motor tasks. We quantified how subjects integrated monocular cues (contour and texture) and binocular cues (disparity and vergence) about 3D surface orientation throughout an object placement task and found that binocular cues contributed more to online control than planning. A temporal analysis of corrective responses to stimulus perturbations revealed that the visuomotor system processes binocular cues faster than monocular cues. This suggests that binocular cues dominated online control because they were available sooner, thus affecting a larger proportion of the movement. This was consistent with our finding that the relative influence of binocular information was higher for short duration movements than long-duration movements. A motor control model that optimally integrates cues with different delays accounts for our findings and shows that cue integration for motor control depends in part on the time course of cue processing. PMID- 15820517 TI - Generic and customized digital image enhancement filters for the visually impaired. AB - This study compares the effectiveness of various image enhancement filters for improving the perceived visibility of coloured digital natural images for people with visual impairment. Generic filters were compared with Peli's adaptive enhancement and adaptive thresholding and custom-devised filters based on each subject's contrast sensitivity loss. Subjects with low vision made within filter rankings followed by between filter ratings. In general, subjects preferred filters with lower gains. Unsharp masking resulted in a significant increase in perceived visibility for some image types (p < or = 0.05) while Peli's adaptive enhancement, edge enhancement and histogram equalization resulted in borderline improvements. Adaptive thresholding and the custom devised filter did not result in overall improvements in perceived visibility. PMID- 15820518 TI - Eccentricity effects on lateral interactions. AB - We attempted to resolve an apparent conflict between the lack of psychophysical evidence of collinear facilitation at the near-periphery and physiological evidence from the monkey showing collinear effects extra-fovealy. We compared collinear and orthogonal configurations to discount facilitation due to reduced positional uncertainty. Detection thresholds were measured for Gabor targets at eccentricities of 0 degrees - 4 degrees, flanked by collinear or orthogonal flankers. Like in previous reports in the literature, results varied among subjects when the stimulus position was off-fixation. We found reduced facilitation at eccentricities as small as 1 degrees - 2 degrees. Moreover, facilitation did not increase when the stimuli were M-scaled or when observers received more practice. However, a larger proportion of subjects showed collinear facilitation when attention was directed to the tested configurations. The results suggest that differences in allocation of attention along the visual field may affect the underlying lateral interactions, consequently resulting in eccentricity effects as well as inter-observer variability. PMID- 15820519 TI - Vertical-disparity gradients are processed independently in different depth planes. AB - We examined the effects of vertical-disparity gradients on apparent depth curvature of textured surfaces. In Experiment 1, vertical disparities induced expected curvatures when the surface had a horizontal disparity of < +/-40.34'. A central row of elements, lacking vertical disparities, ceased to have the same apparent curvature as the surface when the horizontal disparity between row and surface exceeded +/-5'. In Experiment 2, vertical disparities were not pooled between superimposed surfaces separated by horizontal disparities > +/-10'. Thus, vertical-disparity gradients are not pooled over depth for curvature perception. Our results suggest that vertical disparities are used to determine distances to surfaces directly, rather than to estimate vergence. PMID- 15820520 TI - Visually perceived vertical (VPV): induced changes in orientation by 1-line and 2 line roll-tilted and pitched visual fields. AB - We report a series of nine experiments which show that a single roll-tilted line in darkness induces changes of the orientation perceived as vertical (VPV) that are similar in magnitude and direction to those measured by Witkin and Asch (1948a) [Studies in space orientation. I. Perception of the upright with displaced visual fields. Journal of Experimental Psychology, 38, 762-782] with the classical square 4-sided frame, and we describe the configuration-independent mass-action rules by which the influences of the individual lines influences are combined. Clockwise (cw) and counterclockwise (ccw) orientations of a line produce cw and ccw displacements of the VPV setting, respectively, with effect magnitude increasing approximately linearly with line orientation (e.g., a 66.25 degrees - long line at 25 degrees horizontal eccentricity that varies in roll tilt through +/-13.2 degrees around vertical generates a systematic variation in VPV over +/-7 degrees). The slope of the VPV-vs-roll-tilt function increases with line length along a negatively accelerated exponential function (length constant = 17.1 degrees). The influences of two bilaterally symmetric lines combine linearly and algebraically and the combined influence is linearly related to the sum of the VPVs for the 1-line components with a slope equal to 0.91 for short lines and 0.66 for long lines; thus, VPV for short lines manifests nearly complete additive summation, but for long lines, the 2-line VPV is nearer to the average of the VPV values for the two components measured separately. The effectiveness of the conjunction of two line segments within a visual scene does not depend on their separate orientations, only on their sum. Individual lines from pitched-only planes or from combinations of such planes generate identical influences to those generated from lines in frontoparallel planes with the same image orientations at the eye of the observer (their "retinal orientations"). Retinal orientation is the key to the induction of VPV change independently of the line's plane of origin. PMID- 15820521 TI - Exercise might favor reverse cholesterol transport and lipoprotein clearance: potential mechanism for its anti-atherosclerotic effects. AB - Livers of C57 BL/6 mice exercised for 2 weeks showed a dramatic increase in scavenger receptor B1 (SR-B1), CD36 and low density lipoprotein (LDL) receptor and a decrease in acetyl LDL receptor gene expression. These effects on lipoprotein receptors are reminiscent of the effects mediated by peroxisome proliferator-activated receptor (PPARgamma) ligands. PMID- 15820522 TI - Low-dose cholinesterase inhibitors do not induce delayed effects on cerebral blood flow and metabolism. AB - The acetylcholinesterase (AChE) inhibitors sarin and pyridostigmine bromide (PB) have been proposed as causes of neurobehavioral dysfunction in Persian Gulf War veterans. To test possible delayed effects of these agents, we exposed rats to low (subsymptomatic) levels of sarin (0.5 LD50 s.c. 3 times weekly) and/or PB (80 mg/L in drinking water) for 3 weeks. Controls received saline s.c. and tap water. At 2, 4 and 16 weeks after exposure, regional cerebral blood flow (rCBF) and glucose utilization (rCGU) were measured in conscious animals with the Iodo-14C antipyrine and 14C-2 deoxyglucose methods, respectively. Two weeks after exposure, PB+sarin caused significant rCBF elevations, but no changes in rCGU, in neocortex, with lesser effects on allocortex. Four weeks after exposure, the same general pattern was found with sarin. Only a few changes were found at 16 weeks post-treatment. The predominant effects of sarin or PB+sarin on rCBF at earlier times after treatment are consistent with the well known direct cerebral vascular effect of cholinergic agonists. The lack of changes in rCBF and rCGU observed at 16 weeks after treatment does not support the hypothesis that repeat exposure to low-dose cholinesterase inhibitors can generate permanent alterations in cerebral activity. PMID- 15820523 TI - Post-trial treatment with the nicotinic agonist metanicotine: Differential effects in Wistar rats with high versus low rearing activity. AB - Laboratory rats, although identical in strain, sex, age and housing conditions, can differ considerably in behavior and physiology. When screened in an open field, for example, Wistar rats can be assigned to subgroups, based on the measure of rearing activity (high, low rearing activity; HRA/LRA). Such rats have previously been found to differ in dopaminergic and cholinergic brain mechanisms, reactivity to cholinergic drugs, and in tests of learning and memory. Here, we asked whether HRA and LRA rats might respond differently to nicotinic treatment, when given during the consolidation of an aversive experience. Therefore, we tested them for performance in an inhibitory avoidance task where they received post-trial injections of either saline, or the nicotinic agonist metanicotine (RJR-2403, 0.017-1.7 mg/kg, i.p.). In support of previous findings, saline treated LRA rats showed a trend for higher step-in latencies than HRA rats after shock experience. Furthermore, metanicotine was effective only in LRA rats: Compared to their respective saline-treated controls, the retention scores of LRA rats were decreased after post-trial treatment with the highest dose (1.7 mg/kg). Thus, the nicotinic agonist had an amnestic-like effect dependent on dose and subject-dependent factors (HRA/LRA). These findings are discussed with respect to possible drug actions on mnestic and non-mnestic mechanisms, and the importance of taking subject-dependent variability into account when analysing drug effects. PMID- 15820524 TI - Partial serotonergic denervation decreases progenitor cell proliferation in the adult rat hippocampus, but has no effect on rat behavior in the forced swimming test. AB - It has been hypothesized that impaired serotonin (5-HT) neurotransmission contributes to the pathophysiology of depression. Recently, it was shown in adult animals that antidepressants which influence 5-HT levels increase hippocampal progenitor cell proliferation, suggesting an important role of the 5-HT system in the regulation of adult neurogenesis. In this study, we investigated the effects of partial 5-HT denervation of the adult rat forebrain caused by a single para chloroamphetamine (PCA) administration (10 mg/kg, s.c.) on hippocampal progenitor cell proliferation measured by Ki-67 immunohistochemistry over a 3-week time period. Moreover, behavior of rats was analyzed by using the forced swimming test which serves as a so-called animal model of depression. One week after PCA administration, a significant decrease of Ki-67 immunopositive cells and cell clusters (-25% and -53%, respectively) was found which was recovered over the 3 week period. The decrease in progenitor cell proliferation in the dentate gyrus per animal was positively correlated with the 5-HT denervation. Behavioral analysis of rats revealed no significant differences between vehicle and PCA treated animals at any of the examined time points. These findings indicate a regulative role of 5-HT for hippocampal progenitor cell proliferation and neurogenesis, but its relation to animal models of depression remains to be elucidated. PMID- 15820525 TI - Dietary cadmium exposure attenuates D-amphetamine-evoked [3H]dopamine release from striatal slices and methamphetamine-induced hyperactivity. AB - Prolonged exposure to environmentally relevant amounts of CdCl2 results in cadmium accumulation in dopamine-rich brain regions, such as striatum. Exposure to these low levels of cadmium also diminishes cocaine-induced hyperactivity and conditioned reinforcement. The goal of the present study was to assess the effect of cadmium on amphetamine pharmacology. Direct application of cadmium (0.1-100 microM), within the concentrations reported in brain after chronic exposure, to preloaded rat striatal slices did not alter D-amphetamine-evoked [3H]dopamine release. To determine the effect of dietary cadmium exposure on amphetamines, rats received ad libitum access to diet containing CdCl2 (10 or 100 ppm) or to control diet for 30 days and then D-amphetamine-evoked [3H]dopamine release and methamphetamine-induced hyperactivity were measured. Dietary CdCl2 exposure produced a marked increase in cadmium blood and brain levels, approximate to environmental metal exposure. Dietary cadmium exposure was associated with decreased potency of D-amphetamine to evoke [3H]dopamine release. Cadmium-exposed rats were also less sensitive to the locomotor-activating effect of acute methamphetamine (0.3 or 1.0 mg/kg) injection. The present findings demonstrate that the presence of cadmium in brain is not sufficient for the inhibition of D amphetamine-evoked dopamine release. This suggests that cadmium does not directly interfere with the mechanism of action for amphetamine pharmacology; rather, it suggests that long-term cadmium exposure induces a change in the number and/or function of striatal neurons. PMID- 15820526 TI - Adolescent and adult female rats differ in sensitivity to nicotine's activity effects. AB - More than 90% of cigarette smokers begin smoking during adolescence. This between subjects repeated-measures experiment examined: (1) nicotine's acute effects on activity in adolescent and adult female Sprague-Dawley rats (Drug Phase I); (2) the effects of age of initial nicotine exposure on activity when nicotine was not administered (Interim Phase); and (3) the effects of age of initial nicotine exposure on later responses to nicotine (Drug Phase II). The experiment consisted of three separate phases. In Drug Phase I, animals were administered either 0 (saline), 0.01, 0.10, 0.50, or 1.0 mg/kg nicotine via subcutaneous injections for 12 days and horizontal activity was measured daily. During the Interim Phase (no drug phase), activity was measured but nicotine was not administered. During Drug Phase II, the same animals were administered the same nicotine dosages as in Drug Phase I for 12 days and activity was measured daily. Drug Phase I revealed dose response differences between adolescent and adult female rats. In addition, animals initially exposed to nicotine in adolescence exhibited greater sensitivity to nicotine's activity-increasing effects than did females initially exposed to nicotine in adulthood (i.e., Drug Phase II). PMID- 15820527 TI - Strain and sex alter effects of stress and nicotine on feeding, body weight, and HPA axis hormones. AB - Gender and genotype result in differential sensitivity to stress and to nicotine. Male and female Sprague-Dawley and Long-Evans rats exhibit different behavioral responses to immobilization stress and to chronically-administered nicotine, suggesting that these animals may be useful to model human variability in stress and nicotine sensitivity. It is possible that differences in sensitivity of the hypothalamo-pituitary-adrenocortical (HPA) axis might account for these sex and strain differences. This experiment examined corticosterone (CORT) and adrenocorticotropin hormone (ACTH) responses of male and female Sprague-Dawley (n=117) and Long-Evans (n=120) rats administered 0, 6, or 12 mg/kg/day nicotine for 14 days; half of each treatment group was exposed to immobilization stress (20 min/day). Feeding and body weight also were measured. Nicotine increased CORT and ACTH levels of Sprague-Dawley females only. Stress increased CORT and ACTH levels of all groups except for Long-Evans females. Nicotine and stress decreased feeding and body weight with greatest effects in Long-Evans females. CORT, feeding, and body weight were positively correlated among stressed females. These findings suggest that strain differences in HPA axis, body weight, and feeding responses to nicotine and to stress are robust among females but not among males. CORT reactivity and female sex hormones may explain these differences. PMID- 15820528 TI - The effect of zotepine, risperidone, clozapine and olanzapine on MK-801-disrupted sensorimotor gating. AB - Dizocilpine (MK-801; 0.3 mg/kg i.p.)-induced disruption in prepulse inhibition of the acoustic startle response (PPI) can be preferentially restored by "atypical" antipsychotics. In contrast, some findings indicate that not all of the "atypical" antipsychotics, such as clozapine and risperidone, are effective in restoring the NMDA antagonist-induced deficits in PPI. In our study, we evaluated the effect of four different "atypical" antipsychotic drugs on deficits in PPI induced by MK-801. Zotepine and risperidone have high affinities to D2-like and 5 HT2A receptors, while clozapine and olanzapine have multipharmacological profiles with the highest affinities to serotonin 5-HT1A,2A/2C receptors and muscarinic receptors. Results have shown that MK-801 disrupted PPI and increased the ASR in rats. Our results showed no effect of zotepine (1 and 2 mg/kg) and risperidone (0.1 and 1 mg/kg) on disrupted PPI by MK-801. Administration of clozapine (5 and 10 mg/kg) and olanzapine (2.5 and 5 mg/kg) restored the deficits in PPI induced by MK-801. Additionally, we found a decrease of approximately 46% in PPI after administration of clozapine (5 mg/kg) and olanzapine (2.5 and 5 mg/kg) without MK 801 treatment. In summary, the four "atypical" antipsychotics had different efficacies to restore the disrupted PPI by MK-801. Only clozapine and olanzapin restored the MK-801-induced deficits in PPI. PMID- 15820529 TI - The role of the phosphatidyinositol-linked D1 dopamine receptor in the pharmacology of SKF83959. AB - SKF83959, previously described as an antagonist of the D1 dopamine receptor, has been shown to be a potent anti-parkinsonian agent. However, its mechanism of action is unknown. The present communication was designed to study the mechanism by which SKF83959 exerts its pharmacological effects. SKF83959 induced contralateral rotations in the unilateral 6-OHDA-lesioned rat model of Parkinson's disease (PD). The rotations were completely blocked by the D1 dopamine receptor antagonist, SCH23390. The response was not affected by the serotonin receptor antagonist, mesulergine and was transiently attenuated by alpha1 adrenergic or D2 dopamine receptor antagonists, prazosin or spiperone, respectively. Injection of 0.5 and 1 mg/kg SKF83959 elicited significant elevations in IP3 accumulation in lesioned as compared to intact striata. This effect was blocked by SCH23390 at a dose that completely obviated the rotational response to SKF83959, suggesting that activation of the PI-linked D1 dopamine receptor and the PLC/IP3 pathway may be the underlying mechanism for the rotational activity induced by SKF83959. The present data provide the first evidence that the PI-linked D1 dopamine receptor plays a role in regulating motor activity in striatum and that modulation of the D1 dopamine receptor/PLC/IP3 pathway may be a novel target in the discovery of drugs for the treatment of Parkinson's disease. PMID- 15820530 TI - Progesterone inhibits behavioral responses and estrogen increases corticosterone levels after acute cocaine administration. AB - Accumulating evidence suggests that estrogen and progesterone contribute to the sexually dimorphic behavioral response to cocaine. In this study, we tested the hypothesis that varying the level of estrogen or progesterone affects cocaine induced locomotive behavior in female rats. Ovariectomized (OVX) rats received estrogen (0, 5, 10, 20, or 50 microg) 48 h or progesterone (0, 50, 100, 250, or 500 microg) 24 h before acute saline or cocaine (15 mg/kg) administration. Although estrogen did not affect cocaine-induced ambulatory and rearing behaviors, it affected stereotypic behaviors regardless of cocaine administration (animals receiving 50 microg had higher stereotypic counts than did the OVX group). In contrast, progesterone affected rearing activity dose-dependently: 50 and 500 microg of progesterone inhibited, whereas 100 microg and 250 microg stimulated, rearing in response to cocaine. That estrogen and progesterone did not affect overall baseline behavioral activity suggests their effects are mediated in part through interactions with cocaine. Progesterone administration did not affect corticosterone levels in saline- or cocaine-treated rats. Estrogen administration, however, affected levels of corticosterone both at baseline and after cocaine treatment. After accounting for baseline differences, we found that rats receiving 5 or 10 microg of estrogen and cocaine had higher percentage increases in serum corticosterone levels than did the control group that did not receive estrogen. On the basis of these observations, we suggest that progesterone fluctuations during the estrous cycle impact cocaine-induced behavioral responses, whereas estrogen may affect activity in the hypothalamic pituitary-adrenal axis. Thus, dose-dependent effects of gonadal hormones may underlie some of the reported sex differences and reproductive cycle effects of cocaine. PMID- 15820531 TI - The effects of the GABAA antagonist bicuculline on cocaine self-administration in rats exposed to lead during gestation/lactation. AB - The present investigation examined the effects of perinatal lead exposure on cocaine self-administration following a GABAA antagonist pretreatment. Female rats were exposed to either 0 or 16 mg lead daily for 30 days prior to breeding with unexposed males. Beginning on postnatal day (PND) 75, control (N=10) and lead-exposed (N=8) animals were trained to self-administer 0.50 mg/kg cocaine intravenously (IV). After stable responding was established, animals were tested at 0.03 and 0.06 mg/kg cocaine delivered intravenously (IV), combined with intraperitoneal (IP) administration of either saline, 0.50, 1.00 or 2.00 mg/kg bicuculline (a GABAA antagonist). The results showed that control animals increased self-administration responding at a cocaine dose of 0.06 mg/kg as bicuculline dose increased. Lead-exposed animals exhibited an opposite pattern, i.e., a decrease in active (cocaine) lever responding occurred as the bicuculline dose was increased. Results at the 0.03 mg/kg cocaine dose failed to show group separation, or significant changes consequent to the bicuculline pretreatment. The data suggest that GABA antagonism results in increased reward potency of a low dose of cocaine and further, that this effect is differentially expressed in animals exposed to perinatal lead. PMID- 15820532 TI - Reconditioning of drug-related cues: a potential contributor to relapse after drug reexposure. AB - To investigate the process of relapse to drug seeking caused by reexposure to drugs, we studied the consequences of recurring instances of stimuli-drug associations using heroin conditioned place preference (CPP) in rats. After original conditioning and extinction, rats received either a single compartment heroin pairing (reconditioning) or were primed with heroin and tested for reinstatement of CPP. It was found that the session of reconditioning, but not the session of reinstatement, caused the reappearance of a preference for the heroin-paired compartment on a test given 24 h later, in drug-free conditions. The effect of reconditioning was found to be dependent on heroin doses, and was not seen when heroin injections were given outside the conditioning environment. Furthermore, a single session of reconditioning elevated heroin seeking even on a test given 96 h later. Finally, heroin seeking was found to be significantly elevated on a test given 28 days after the last extinction session whether animals received 1 or 3 reconditioning sessions. These results suggest that the motivational value of cues associated with heroin is not eliminated by extinction and, importantly, that these cues can rapidly regain their ability to promote drug seeking behavior if they are re-associated with the effect of heroin. PMID- 15820533 TI - Metyrapone and cocaine: a double-blind, placebo-controlled drug interaction study. AB - Pre-clinical research suggests that suppression of adrenocorticosteroid synthesis might decrease susceptibility to stress-induced relapse. Metyrapone effectively suppresses cortisol synthesis and thus might have promise as a cocaine dependence treatment. The present inpatient study evaluated the interaction of metyrapone and cocaine to assess the safety of conducting an outpatient trial. Twelve nontreatment-seeking cocaine-dependent individuals completed this double-blind, placebo-controlled, crossover study with two factors: medication (750 mg of metyrapone vs. placebo) and infusion (40 mg of cocaine vs. saline). Safety measures included vital signs, adverse events, and electrocardiogram. Efficacy measures included visual analog scale (VAS) ratings of craving and drug effect. Neuroendocrine measures included cortisol and ACTH. As predicted, metyrapone was well tolerated and did not exacerbate cocaine's physiological effects. Also as predicted, metyrapone did not significantly alter cocaine's subjective effects. The results of the present study suggest that metyrapone at the dose studied can likely be used safely in an outpatient study with active cocaine users. PMID- 15820534 TI - NMDA antagonist modulation of morphine antinociception in female vs. male rats. AB - NMDA antagonists may be useful for their potential to increase or prolong opioid analgesia while attenuating the development of opioid tolerance and dependence. The purpose of this study was to determine whether there are sex differences in NMDA antagonist modulation of morphine antinociception. Adult female and male Sprague-Dawley rats were injected s.c. with saline or one dose of MK-801 (0.005, 0.01, 0.02, or 0.04 mg/kg), dextromethorphan (5, 10, or 20 mg/kg), or LY235959 (0.5, 1.0, or 2.0 mg/kg) in combination with saline or one dose of morphine (1.8, 3.2, or 5.6 mg/kg), and tested on the 50 degrees C hotplate and tail withdrawal assays 15-120 min post-injection. At the doses examined, only LY235959 produced any antinociception when administered alone. MK-801 attenuated morphine antinociception on both assays, but only at sporadic (inconsistent) dose combinations. Dextromethorphan increased morphine antinociception on the hotplate but not tail withdrawal assay, at all three morphine doses in males, but only the higher morphine doses in females. In contrast, LY235959 modulated morphine antinociception on both assays; the lowest dose attenuated, and higher doses enhanced morphine antinociception, but the particular morphine doses and assay in which these effects occurred depended on the sex of the subject. Thus, all three NMDA antagonists modulated morphine antinociception in female and male rats, but the direction of this modulation depended on the particular antagonist examined, the nociceptive test, the dose of antagonist and of morphine, and time post injection. PMID- 15820535 TI - Getting prevention right: maintaining the status quo is not an option. PMID- 15820536 TI - Assessing home visiting programs to prevent child abuse: taking silver and bronze along with gold. PMID- 15820537 TI - Rethinking healthy families: a continuous responsibility. PMID- 15820538 TI - Clarifying the impact of the Nurse-Family Partnership on child maltreatment: response to Chaffin (2004). PMID- 15820539 TI - Response to Chaffin (2004). PMID- 15820540 TI - Response to Chaffin (2004). PMID- 15820542 TI - The influence of support and stress on maternal attitudes. AB - OBJECTIVES: This study examines an expanded version of Belsky's (1984) multi dimensional process model of parenting to determine whether changes in stress and support influence maternal attitudes during the first year of a child's life. METHOD: Data were collected from mothers of newborns eligible for Hawaii's Healthy Start program who had been randomly assigned to home visitation (n=108) or control (n=104) status. Multiple regression analyses were used to test hypotheses regarding the influence of change in contextual sources of support and stress on parenting attitudes as measured by the Child Abuse Potential Inventory (CAPI) Abuse Scale. A post hoc analysis of variance was conducted to explore interactions among the predictor variables and home visitation. RESULTS: Stress in the form of change in public assistance status led to more punitive parenting attitudes while increased support from the partner and from close associates emerged as significant predictors of less punitive parental attitudes. The exploratory analyses suggest that home visitation may moderate the impacts of stress and support on maternal attitudes. CONCLUSIONS: In line with Belsky's (1984) process model of parenting, increased stress adversely impacted maternal attitudes regarding physical punishment while increased support exerted favorable effects. The results also support further elaboration of Belsky's model including the expansion of marital support to include unmarried partners and the need to consider the impact of formal support sources on parenting. PMID- 15820543 TI - Predicting long-term outcomes for women sexually abused in childhood: contribution of abuse severity versus family environment. AB - OBJECTIVE: Child sexual abuse (CSA) has been associated with adverse adult psychosocial outcomes, although some reports describe minimal long-term effects. The search for explanations for the heterogeneous outcomes in women with CSA has led to an examination of a range of CSA-related factors, from the severity of individual CSA incidents to the childhood family environment. This study compares three factors for predicting adult outcomes: a multidimensional CSA Severity Scale, the presence or absence of CSA, and family environment. METHODS: The effect of CSA on adult outcomes was examined among 290 community-dwelling women raised in intact families. Standardized measures and a focused interview were used to collect data, with siblings as collateral informants. RESULTS: Comparison of a multidimensional CSA Severity Scale to a dichotomous measure of the presence or absence of CSA showed that the Severity Scale did not have greater predictive value for adult outcomes than the dichotomous measure, nor was it as parsimonious. Childhood family environment scales added significantly to the predictive ability of the dichotomous measure. CONCLUSION: The specific characteristics of a CSA experience may be less important than the occurrence of CSA and the family environment of women for predicting long-term outcomes. PMID- 15820544 TI - Higher prevalence of childhood sexual abuse among Latino men who have sex with men than non-Latino men who have sex with men: data from the Urban Men's Health Study. AB - OBJECTIVE: The prevalence of childhood sexual abuse among Latino adult men who have sex with men who live in the US was estimated because a history of childhood sexual abuse increases HIV sexual risk behaviors and other negative health outcomes in adulthood. METHOD: The Urban Men's Health Study is a random-digit telephone probability survey of 2881 adult men who have sex with men (MSM) aged 18 years or older residing in San Francisco, New York, Los Angeles, and Chicago. Self-reported history of childhood sexual abuse was the main outcome measure gathered from 2692 MSM. RESULTS: A significantly higher proportion of Latino MSM reported sexual abuse before age 13 (22%) than did non-Latino MSM (11%). CONCLUSIONS: Latino MSM are twice as likely to report a history of childhood sexual abuse than are non-Latino MSM. Given the association between childhood sexual abuse and increased risk for HIV and other negative health outcomes, health providers must remain vigilant to the possibility of childhood sexual abuse histories among their Latino patients. PMID- 15820545 TI - Neurobiology of emotion and high risk for schizophrenia: role of the amygdala and the X-chromosome. AB - Abnormalities in emotion processing and in structure of the amygdala have consistently been documented in schizophrenia. A major question is whether amygdala abnormalities reflect a genetic vulnerability for the disease. In the present paper, we reviewed Magnetic Resonance Imaging (MRI) studies that reported amygdala measures in several high-risk populations: subjects from the general population with subclinical schizophrenia symptoms and relatives of schizophrenia patients. In addition, we reviewed the evidence regarding Klinefelter syndrome (characterised by an additional X-chromosome), which has also been related to an increased risk for schizophrenia. Overall, the evidence points to structural abnormalities of the amygdala in individuals at increased risk for schizophrenia. Although the genetic basis of amygdala deficits remains unclear, abnormalities (of genes) on the X-chromosome might play a role as suggested by the evidence from individuals with sex chromosome aneuploidies. We propose that amygdala abnormalities are an endophenotype in schizophrenia and may account for subtle emotional processing deficits that have been described in these high-risk groups. PMID- 15820546 TI - The neuropsychology of obsessive compulsive disorder: the importance of failures in cognitive and behavioural inhibition as candidate endophenotypic markers. AB - Obsessive compulsive disorder (OCD) is a highly debilitating neuropsychiatric condition with estimated lifetime prevalence of 2-3%, more than twice that of schizophrenia. However, in contrast to other neuropsychiatric conditions of a comparable or lesser prevalence, relatively little is understood about the aetiology, neural substrates and cognitive profile of OCD. Despite strong evidence for OCD being familial, with risk to first-degree relatives much greater than for the background population, its genetic underpinnings have not yet been adequately delineated. Although cognitive dysfunction is evident in the everyday behaviour of OCD sufferers and is central to contemporary psychological models, theory-based studies of neurocognitive function have yet to reveal a reliable cognitive signature, and interpretation has often been confounded by failures to control for co-morbidities. The neuroimaging findings in OCD are amongst the most robust reported in the psychiatric literature, with structural and functional abnormalities frequently reported in orbitofrontal cortex, anterior cingulate cortex, and caudate nucleus. In spite of this, our relative lack of understanding of OCD neurochemical processes continues to impede progress in the development of novel pharmacological treatment approaches. Integrating the neurobiological, cognitive, and clinical findings, we propose that OCD might usefully be conceptualised in terms of lateral orbitofrontal loop dysfunction, and that failures in cognitive and behavioural inhibitory processes appear to underlie many of the symptoms and neurocognitive findings. We highlight existing limitations in the literature, and the potential utility of endophenotypes in overcoming these limitations. We propose that neurocognitive indices of inhibitory functions may represent a useful heuristic in the search for endophenotypes in OCD. This has direct implications not only for OCD but also for putative obsessive-compulsive spectrum conditions including attention deficit hyperactivity disorder, Tourette's syndrome, and trichotillomania (compulsive hair pulling). PMID- 15820547 TI - Imprinted gene expression in the brain. AB - In normal mammals, autosomal genes are present in duplicate (i.e. two alleles), one inherited from the father, and one from the mother. For the majority of genes both alleles are transcribed (or expressed) equally. However, for a small subset of genes, known as imprinted genes, only one allele is expressed in a parent-of origin dependent manner (note that the 'imprint' here refers to the epigenetic mechanism through which one allele is silenced, and is completely unrelated to classical 'filial imprinting' manifest at the behavioural level). Thus, for some imprinted genes expression is only (or predominantly) seen from the paternally inherited allele, whilst for the remainder, expression is only observed from the maternally inherited allele. Early work on this class of genes highlighted their importance in gross developmental and growth phenotypes. Recent studies in mouse models and humans have emphasised their contribution to brain function and behaviour. In this article, we review the literature concerning the expression of imprinted genes in the brain. In particular, we attempt to define emerging organisation themes, especially in terms of the direction of imprinting (i.e. maternal or paternal expression). We also emphasise the likely role of imprinted genes in neurodevelopment. We end by pointing out that, so far as discerning the precise functions of imprinted genes in the brain is concerned, there are currently more questions than answers; ranging from the extent to which imprinted genes might contribute to common mental disorders, to wider issues related to how easily the new data on brain may be accommodated within the dominant theory regarding the origins and maintenance of imprinting, which pits the maternal and paternal genomes against each other in an evolutionary battle of the sexes. PMID- 15820548 TI - Applications of prism adaptation: a tutorial in theory and method. AB - Data and theory from prism adaptation are reviewed for the purpose of identifying control methods in applications of the procedure. Prism exposure evokes three kinds of adaptive or compensatory processes: postural adjustments (visual capture and muscle potentiation), strategic control (including recalibration of target position), and spatial realignment of various sensory-motor reference frames. Muscle potentiation, recalibration, and realignment can all produce prism exposure aftereffects and can all contribute to adaptive performance during prism exposure. Control over these adaptive responses can be achieved by manipulating the locus of asymmetric exercise during exposure (muscle potentiation), the similarity between exposure and post-exposure tasks (calibration), and the timing of visual feedback availability during exposure (realignment). PMID- 15820549 TI - The use of norms of reaction to analyze genotypic and environmental influences on behavior in mice and rats. AB - Norms of reaction (NoRs) represent the phenotypic values of genotypes as functions of environmental parameters and permit the visualization of differences in phenotypic response of different genotypes. NoR graphs can be used to analyze interactions between genotypic and environmental factors during development to produce phenotypes in inbred strains of rats and mice. We describe the main features of NoRs, the history of their use in this context, and discuss several applications in behavioral neuroscience. In addition, we give a test for determining whether distinct strains have different NoRs. PMID- 15820550 TI - Effects of appraisal and coping on the neuroendocrine response to extreme stress. AB - Although many people are exposed to extreme stress, only some of them develop psychobiological disturbances that can lead to posttraumatic stress disorder (PTSD) or other posttrauma psychopathology. This paper examines the effects of different types of appraisal and coping to find clues to how individuals differ in their neuroendocrine responses to extreme stress. It proposes a conceptual model for components of the adult response to stressors. Threat appraisal and defensive coping may play crucial roles in determining the neuroendocrine response to trauma with potential mental health consequences, particularly PTSD. PMID- 15820551 TI - Measuring emotional processes in animals: the utility of a cognitive approach. AB - Contemporary researchers regard emotional states as multifaceted, comprising physiological, behavioural, cognitive and subjective components. Subjective, conscious experience of emotion can be inferred from linguistic report in humans, but is inaccessible to direct measurement in non-human animals. However, measurement of other components of emotion is possible, and a variety of methods exist for monitoring emotional processes in animals by measuring behavioural and physiological changes. These are important tools, but they have limitations including difficulties of interpretation and the likelihood that many may be sensitive indicators of emotional arousal but not valence pleasantness/unpleasantness. Cognitive components of emotion are a largely unexplored source of information about animal emotions, despite the fact that cognition-emotion links have been extensively researched in human cognitive science indicating that cognitive processes-appraisals of stimuli, events and situations-play an important role in the generation of emotional states, and that emotional states influence cognitive functioning by inducing attentional, memory and judgement biases. Building on this research, it is possible to design non linguistic cognitive measures of animal emotion that may be especially informative in offering new methods for assessing emotional valence (positive as well as negative), discriminating same-valenced emotion of different types, identifying phenotypes with a cognitive predisposition to develop affective disorders, and perhaps shedding light on the issue of conscious emotional experiences in animals. PMID- 15820552 TI - Vagal nerve stimulation: a review of its applications and potential mechanisms that mediate its clinical effects. AB - Vagal nerve stimulation (VNS) is an approved treatment for epilepsy and is currently under investigation as a therapy for other disorders, including depression, anxiety and Alzheimer's disease. This review examines the pre clinical and clinical literature relating to VNS. A brief historical perspective is given, followed by consideration of the efficacy of the various clinical applications of VNS. Finally, what is known about the mechanism by which VNS exerts clinical benefit is considered. It is concluded that although the precise mechanism of action of VNS is still unknown, the search for the mechanism has the potential to lend new insight into the neuropathology of depression. It is important that prior assumptions about the influence of VNS on particular aspects of brain function do not constrain the investigations. PMID- 15820554 TI - Tight junctions, from tight intercellular seals to sophisticated protein complexes involved in drug delivery, pathogens interaction and cell proliferation. AB - Tight junctions (TJ) are intercellular seals that regulate the passage of ions and molecules through the paracellular pathway. This property allows epithelial and endothelial sheets to function as barriers between different body compartments. Although TJ were identified by electron microscopy since the 1950's, the analysis in recent years of their molecular composition, has depicted TJ as signaling complexes involved in a wide spectrum of physiological and pathological processes, such as cell growth and differentiation, cancer and metastasis, and pathogen interaction. This new knowledge has in turn given rise to novel strategies for enhancing the paracellular flux of poorly absorbed therapeutics. PMID- 15820555 TI - Molecular perspective on tight-junction assembly and epithelial polarity. AB - Apical-basal polarity and a highly organized actin cytoskeleton are main characteristics of epithelial cells that support exchange of ions and nutrients from one body compartment to another. The junctional complexes, localized to the apical end of the basolateral domain of the plasma membrane, are not simply epithelial barriers in paracellular transport or fences preventing diffusion of integral proteins in the plasma membrane, but also contain proteins involved in the maintenance of the physiologic epithelial cell state and signal transduction. Claudin-based tight junctions and E-cadherin-based adherens junctions have been extensively studied. Nectins, along with a unique scaffolding protein, afadin, form homophilic and heterophilic trans-dimers and play a key role in identifying cell partners in the primordial cell-cell adhesion. Nectin-based cell-cell adhesion participates in the epithelial morphogenesis, both independently and cooperatively with claudin-based tight junctions and cadherin-based adherens junctions. This review discusses how these adhesion systems interact with each other to form apical junctional complexes, and how they reorganize the actin cytoskeleton in a multistage process of cell adhesion, migration, and polarization. PMID- 15820556 TI - The JAM family of proteins. AB - Junctional adhesion molecules (JAMs) are immunoglobulin superfamily (IgSF) proteins expressed at cell junctions in epithelial and endothelial cells as well as on the surface of leukocytes, platelets, and erythrocytes. Evidence suggests JAM proteins are important for a variety of cellular processes, including tight junction assembly, leukocyte transmigration, platelet activation, angiogenesis and virus binding. This review will highlight what is currently known about the structure, function, and mechanism of action of JAM proteins as well as comment on potential directions for future JAM-related research. PMID- 15820557 TI - CAR: a virus receptor within the tight junction. AB - The coxsackievirus and adenovirus receptor (CAR) mediates cell attachment and infection by coxsackie B viruses and by a number of adenoviruses. CAR also mediates homotypic intercellular interactions. In polarized epithelial cells, CAR is closely associated with the tight junction, where it contributes to the barrier to paracellular flow of solutes and macromolecules. CAR's biological roles are not well defined, but emerging evidence suggests that it may function during embryonic development and in regulating cell proliferation. PMID- 15820558 TI - Occludin: structure, function and regulation. AB - Epithelial and/or endothelial barriers play a critical role in animal, including human, life forms. The tight junction (TJ) is an essential component of these barriers. Occludin is a major component of the TJ. The structure of occludin, including its gene splice variants and protein essential components have been elucidated. Phosphorylation/dephosphorylation plays a major role in regulation of occludin and TJ. Disruption of occludin regulation is an important aspect of a number of diseases. Strategies to prevent and/or reverse occludin downregulation may be an important therapeutic target. PMID- 15820559 TI - Role of claudins in tumorigenesis. AB - The family of more than 20 claudin (CLDN) proteins comprises one of the major structural elements within the apical tight junction apparatus, a dynamic cellular nexus for maintenance of a luminal barrier, paracellular transport, and signal transduction. Loss of normal tight junction functions constitutes a hallmark of human carcinomas. CLDN1 may support tumor suppressive functions in tissues such as the brain, where dramatic loss of expression has been demonstrated in glioblastoma multiforme. The role(s) for CLDNs 3 and 4 in tumorigenesis is less clear. CLDN4 appears to be over-expressed in ovarian and pancreatic carcinomas, and this raises the possibility that a unique, potentially non-toxic cancer cell target may be developed through the design of enterotoxin analogues. Future goals include understanding the biochemical and physiological mechanisms that are perturbed as a consequence of CLDN alterations in the progression of solid tumors. PMID- 15820560 TI - [Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) coinfection: epidemiology, severity and recent therapeutic strategies]. PMID- 15820561 TI - [Incidence and risk factors for thrombosis of central venous catheters in oncology patients]. PMID- 15820562 TI - [Symptomatic thrombosis in central venous catheter in oncology: a predictive score?]. AB - BACKGROUND: Central venous catheters are essential to management of cancer patients. Thrombotic complications are potentially severe, but it is difficult to prescribe systematically a prophylactic treatment. So it is necessary to identify the higher risk patients who need a prophylaxis. AIM: To identify factors associated with the development of clinically significant venous thrombosis in cancer patients with long-term catheters. METHODS: Monocentric prospective study about 5447 long-term central venous catheters inserted into patients receiving treatment for solid tumours (50% of breast cancers). Clinically significant catheter-related thromboses are confirmed by ultrasonography, phlebography or scanner. RESULTS: The median duration of catheter use is 147 days. There are 135 clinically significant catheter-related thromboses. The risk for thrombosis is 0.1149 events per 1000 device days. The incidence of symptomatic thrombosis is 0.9% at 30 days, 1.36% at 60 days, 1.83% at 90 days and 2.25% at 120 days. The multivariate analysis shows that female sex, duration of insertion procedure (more than 25 minutes) and place of insertion (femoral place) are factors associated with clinically significant venous thrombosis. The right subclavian insertion causes less risk. The catheters with their tip too "high" in the superior vena cava are systematically changed, so that the position of the catheter tip does not appear as a risk factor in our study. The disease stage and the type of treatments are not well examined. CONCLUSIONS: A predictive score can be made with the three risk factors that have been identified. Thanks to this score it is possible to determine the patients with higher risk for clinically significant catheter-related venous thrombosis. These patients must be more frequently watched over and must receive a prophylactic treatment. The best prophylaxis has to be determined. PMID- 15820563 TI - [Efficacy and safety of pegylated-interferon plus ribavirin in HIV-infected patients co-infected with hepatitis C virus in clinical practice: a 32 cases observational follow-up]. AB - OBJECTIVE: To describe efficacy and safety in clinical practice of pegylated interferon plus ribavirin (INFpeg-Riba) in the treatment of hepatitis C viral infection (HCV) in HIV infected patients. METHODS: Monocentric retrospective study with inclusion of all patients who received at least once INFpeg-Riba before April 1st 2003. All patients were followed up to six months after the end of HCV therapy. RESULTS: Thirty two HIV-positive patients (23 men and 9 women) with chronic hepatitis C treated by INFpeg-Riba were included. The mean age was 43 years. Fourteen patients carried HCV genotype 2 or 3 (43 %) and 18 patients carried genotype 1 or 4 (57%). The Metavir score of fibrosis showed fibrosis F1 (N =3), F2 (N =14), F3 (N =7) and F4 - cirrhosis (N =8). Twenty six patients (81%) were naive for anti hepatitis C drugs. Thirty one per cent of patients were at AIDS stage and 84% were receiving antiretroviral drugs. The mean CD4 cell count was 469 /ml and the plasma RNA HIV was less than 50 copies /ml in 57% of the cases. Adverse events leading to reduction of dose of drugs occurred in 40% and adverse events leading to discontinuation treatment occurred in 12%. A decline of CD4 cell count <200 CD4/ml was observed in 15%. Clearance of HCV-RNA in end of treatment was seen in 46 % and sustained virological response in 34 %. The main predictors of sustained virological response were HCV genotype 2 or 3 (P =0.04) and plasma HIV RNA less than 50 copies/ml (P =0.001). The predictive value of good virological response of a CD4 cell count >350/ml before treatment was very near the statistical significancy (p =0.07). CONCLUSIONS: The efficacy of pegylated interferon plus ribavirin in HIV-HCV co-infected patients is disappointing mainly due to a poor tolerance. In addition to HCV genotype, plasma HIV RNA level and CD4 cell count were essential to predict INFpeg-Riba response and should be taken into account in the process leading to the initiation of such therapy in HIV-HCV co-infected patients. PMID- 15820564 TI - [Primary localized amyloidosis of the urinary tract. A case series of five patients]. AB - PURPOSE: To describe the clinical and radiographic features of patients with primary localized amyloidosis of the urinary tract. METHODS: We report a case of localized amyloidosis of the ureters and bladder. The medical records of four other cases from the French Register of localized amyloidosis were reviewed. RESULTS: The mean age of three men and two women was 53 years. All patients presented with gross hematuria, four patients presented with renal colic, only one patient had irritative lower urinary tract symptoms. Ureter and bladder were involved in three patients, both ureters in two patients and the bladder only, in one patient. Clinical and radiographic presentations mimicked a neoplasia excluded by histologic analysis. Immunohistochemical study was performed in only two cases and revealed lambda light chain amyloidosis. The median follow-up was eight years. Various treatments were performed, and recurrences occurred in two cases. None of the five patients developed monoclonal gammapathy or systemic amyloidosis. CONCLUSION: Primary localized amyloidosis of the urinary tract is a rare disorder and can easily be confused with a neoplasm. The physiopathology is unknown, the prognosis is usually good. There is no specific treatment, and repeated work-up for systemic amyloidosis is unnecessary as local recurrences appear to be the main complication. PMID- 15820565 TI - [Genetic factors predisposing to fibrosis in systemic sclerosis]. AB - PURPOSE: Physiopathology of systemic sclerosis includes autoimmunity factors, endothelial lesions and abnormal fibrotic process which characterizes this disease in the field of systemic autoimmune disorders. Genetic factors of susceptibility are showed by possibility of familial forms of the disease, Choctaw American Indians homogenous population with high disease prevalence of systemic sclerosis and experimental animal models. KEY POINTS: We propose a review of the articles published to date in the literature concerning genetical analysis of genes coding for factors potentially involved in the fibrotic process of systemic sclerosis. This includes cytokines (TNF-alpha, interleukin-1, chemokines), growth factors (TGF-beta), extracellular matrix proteins (collagen, fibrillin, fibronectine) and agents acting on vascular tone (angiotensin converting enzyme and NO synthase). PERSPECTIVES: Identification of genetic factors involved in the susceptibility to fibrosis of systemic sclerosis would lead to a better understanding of physiopathological mechanisms of this disease and to therapeutic targets using immunomodulation with drugs, such as already performed in rheumatoid arthritis. PMID- 15820566 TI - [Treatment of cancer and hematological malignancy in elderly people (Part II)]. AB - PURPOSE: Fifty percents of cancer arise in people older than 65-year-old. Most clinical trials in cancer treatment are limited in patients younger than 65-year old. We review literature-describing particularity of cancer treatment in elderly patients. CURRENT KNOWLEDGE AND KEY POINTS: Therapeutic decisions should be based on an estimation of the patient's life expectancy, and risks and benefits should be weighted up accordingly. Geriatric oncology is made of a geriatric evaluation of patient and of knowledge of clinical trial about elderly patients. FUTURE PROSPECTS AND PROJECTS: We present in this issue the principle of geriatric evaluation and the results of recent clinical trial on elderly cancer patients. PMID- 15820567 TI - [Advances in iron metabolism: a transition state]. AB - PURPOSE: Advances towards the understanding of gene regulation and protein function recently discovered through iron metabolism disorders are the subject of this review. CURRENT KNOWLEDGE AND KEY POINTS: Within a few years the discovery of genes that determine heritable defects of cellular iron uptake or regulation in mice as in humans have provided new insights for investigation into iron metabolism pathways. FUTURE PROSPECTS AND PROJECTS: It is still unclear how connections are made between new proteins in iron uptake, trafficking and regulation of iron homeostasis. Gene expression studies using microarrays technology in different iron conditions should help to explore iron homeostasis further. PMID- 15820568 TI - [Guillain-Barre syndrome and pregnancy]. PMID- 15820569 TI - [Meningeal febrile syndrome and hemiplegia revealing infectious endocarditis: the story of a catheter-related infection]. AB - INTRODUCTION: We report the case of a 48-years patient who is under chimotherapy because of a breast cancer. She was initially found at her place in coma with hyperthermia and neurological deficiency signs. The clinic history and the paraclinical investigations have enabled to detect a catheter-related infection (CRI) as soon as the diagnosis of disseminated infectious endocarditis was made. The resonance imaging confirmed the cerebral blow when it found suspicious images of septic embolus. EXEGESIS: The CRI are usually detected at an early stage and if the classical association CRI-infectious endocarditis-systemical embolus is well known, the discovery on a CRI not treated at the stade of a febril coma is exceptional. CONCLUSION: More than ever the taking care of infectious endocarditis, still delicate and sometimes perilous, requires a global taking care of the patient. PMID- 15820570 TI - [Immediate allergy to oral corticosteroids]. AB - INTRODUCTION: Corticosteroids are widely used for the management of many diseases because of anti-inflammatory and immunomodulatory properties. Sulfite intolerance is well known but immediate hypersensitivity to corticosteroids is rarer and must be reported. EXEGESIS: We report a case of a 38-year-old man, followed since 1986 for asthma with sulfite intolerance. He twice developed urticaria after ingesting betalactam antibiotics associated with corticosteroids of the group A, according to the Matura and Goossens's classification. The allergy inquiry confirmed not only a beta-lactame allergy but also in corticosteroids of the group A. Intradermal reactions with methylprednisolone and hydrocortisone were positive at 7 mm for methylprednisolone on 1/1000 dilution and at 4 mm for hydrocortisone on 1/10 dilution. Thus immediate allergy to corticosteroids was confirmed. But those skin tests were negative for corticosteroids of group B and C. In order to propose an alternative therapy for acute asthma, an intravenous injection of 4 mg of betamethasone was performed without inducing immediate and delayed allergic reactions. CONCLUSION: Corticosteroid allergy is rare and can be fatal. Many asthmatic or acetylsalicylic acid intolerant patients may be concerned. The diagnostic of corticosteroid allergy have to be confirmed by clinical history and skin tests of the different groups to try to identify one or more allergens and to propose a alternative treatment if necessary. PMID- 15820571 TI - [Ganglion cyst at the spinoglenoid notch. A case report and literature review]. AB - INTRODUCTION: Entrapment of the suprascapular nerve at the spinoglenoid notch due to a ganglion cyst is an uncommon event and remains a challenge for clinicians. EXEGESIS: We describe the findings in a 35-year-old patient with nonspecific pain in the shoulder and in whom MR revealed a ganglion cyst in the spinoglenoid notch, confirmed surgically. CONCLUSION: Entrapment of the suprascapular nerve at the spinoglenoid notch due to a ganglion cyst must be considered in the differential diagnosis of nonspecific shoulder pain. MR imaging and electromyography can reach a positive, topographic, and etiologic diagnosis of suprascapular neuropathy. PMID- 15820572 TI - [Acute splenic sequestration: a rare and severe complication of sickle cell anemia in adults. A case report]. AB - INTRODUCTION: Acute splenic sequestration, a well-recognized complication of sickle cell syndromes, is characterized by a sudden decrease in haemoglobin concentration and marked painless splenomegaly. We report a case illustrating the outcome and the treatment options of this complication. CASE REPORT: A 45-year old homozygous woman developed acute splenic sequestration with severe anemia. Red blood cells transfusion led to transient improvement but a relapse-required splenectomy. Long-term outcome was favorable. CONCLUSION: Acute splenic sequestration is a severe complication mainly observed in children. Despite the severity of this complication, prompt diagnosis and appropriate therapy, and particularly red blood cells transfusions, led to a complete recovery. Splenectomy is required in the more severe form of the disease. PMID- 15820573 TI - [Floating aortic thrombosis: an uncommon cause of systemic emboli]. PMID- 15820574 TI - [Pain, dyspnea, hypoxia due to gastric diverticulum]. PMID- 15820575 TI - [Mesenteric panniculitis and common variable immunodeficiency]. PMID- 15820576 TI - [Granulomatous cutaneous manifestation of cytomegalovirus in an immunocompetent patient]. PMID- 15820578 TI - Complexity theory and geographies of health: a critical assessment. AB - The interest of social scientists in complexity theory has developed rapidly in recent years. Here, I consider briefly the primary characteristics of complexity theory, with particular emphasis given to relations and networks, non-linearity, emergence, and hybrids. I assess the 'added value' compared with other, existing perspectives that emphasise relationality and connectedness. I also consider the philosophical underpinnings of complexity theory and its reliance on metaphor. As a vehicle for moving away from reductionist accounts, complexity theory potentially has much to say to those interested in research on health inequalities, spatial diffusion, emerging and resurgent infections, and risk. These and other applications in health geography that have invoked complexity theory are examined in the paper. Finally, I consider some of the missing elements in complexity theory and argue that while it is refreshing to see a fruitful line of theoretical debate in health geography, we need good empirical work to illuminate it. PMID- 15820579 TI - The 'geneticisation' of heart disease: a network analysis of the production of new genetic knowledge. AB - Genetic science is making ever-expanding claims about the (mal)functioning of the body. The 'geneticisation' of health and medicine is extending from rare single gene conditions to more common multi-factorial disease, such as heart disease. The dominant behavioural and socio-spatial explanations of heart disease are now being challenged by genetic claims of deterministic biological causes. This paper builds an account of the transformation of heart disease in the new genetics era, by applying actor network theory (ANT) to the production of genetic knowledge of one aspect of heart disease-hypertension-within a medical genetics laboratory in Glasgow, Scotland. Using this approach, the paper shows that there is no straightforward geneticisation of heart disease. Instead, there is a contested, complex and uncertain understanding of heart disease as genetic, a product of the many people, technologies, natural elements and spaces involved in the network of genetic science knowledge making. The paper concludes that a 'critical' ANT could be developed that acknowledges the inherent unevenness of the network, and connects genetic and socio-spatial explanations of heart disease. PMID- 15820580 TI - Replacing positivism in medical geography. AB - Revisiting debates about philosophical approaches in medical geography suggests that logical positivism may have been prematurely discarded. An analysis of authoritative texts in medical geography and their sources in human geography shows that logical positivism has been conflated with Comtean positivism, science, empiricism, quantification, science politics, scientism and so on, to produce the "standard version" of the all-purpose pejorative "positivism", which it is easy to dismiss as an evil. It is argued that the standard version fails to do justice to logical positivism, being constructed on sources which are at some distance from the logical positivist movement itself. An alternative approach is then developed, an historically and geographically situated interpretation of logical positivism as a deliberately and knowingly constructed oppositional epistemology within an oppressive and anti-scientific culture predicated on idealist intuitionism. Contrasting the standard version with this alternative reading of logical positivism suggests that much may have been lost in human, and thus, medical geography, by throwing out the logical positivist baby with the "positivism" bath water. It is concluded that continuing to unpack the standard version of logical positivism may identify benefits from a more nuanced appreciation of logical positivism, but it is premature to take these to the level of detailed impacts on the kinds of medical geographies that could be done or the ways of doing them. PMID- 15820581 TI - Everyday health histories and the making of place: the case of an English coastal town. AB - During recent years, health geography has focused increasingly on how people's experiences of health and places are interrelated. This has included historical research on a variety of well-known places and remarkable events. In contrast, relatively little attention has been paid to how health and health care can be influential historically to the development, image and identity of 'everyday' and ordinary places. We focus on the case of a small town in Southern England to demonstrate the long and diverse historical connections that exist between health and such places. Using both historical documents and local literature, the paper reviews early invasions and incursions, epidemics of infectious disease, the development of a therapeutic seaside resort, the impacts of the Second World War, and more recent social and economic developments associated with an ageing population. Complex and diverse health-related histories are shown to be interwoven with local and extra-local histories including those of national or international significance. We argue that health histories can be integral to shaping the past and present of places and our broader understandings and interpretations of them. From a disciplinary perspective, we contend that by adopting theoretical perspectives from cultural geography, an enhanced historical geography of health could disentangle these historical strands, tell these important stories and put everyday health histories in their place. PMID- 15820582 TI - Defining regions for locality health care planning: a multidimensional approach. AB - The increasing significance of the role of the general practitioner (GP) in the British National Health Service, evolving from a provider to purchaser and now a key player in the organisation of Primary Care Groups, suggests the need for GPs to possess more and more information about their registered population. GP catchment areas, though an essential basis for providing GPs with important information such as levels of accessibility to surgery, are rarely clearly or accurately defined. Previous approaches towards the definition of GP catchments have been confined to single regionalisation methods, such as mean distance measures, and are prone to problems of either overestimating or underestimating medical service areas. This problem is compounded by a lack of acknowledgement that the application of contrasting catchment methodologies to a common service population has the potential to yield vastly different results which can have serious implications for health care planning and resource allocation. The lack of sophistication in the definition of medical service areas calls for a new methodology to be considered. In this paper, attention is given to the adaptation of multidimensional regional analytical techniques developed outside the health domain and applied in a Regional Health Authority in Northern Ireland. The technique involves the creation of a Synthetic Data Matrix (SDM) which compares patient to GP flow (affiliation) information aggregated at the Census Enumeration District level across a number of catchment areas created using different methodologies. The SDM is then analysed using a modified version of the European Regionalisation Algorithm to create an optimal set of non-overlapping regions according to pre-defined population size and self-containment criteria. The results, a set of compact, robust and highly self-contained catchments, are extremely encouraging. The paper considers the future potential use of such a methodology for health care planning and highlights areas for further research in this field. PMID- 15820583 TI - Zone design for environment and health studies using pre-aggregated data. AB - Many environment and health studies employ geographical areas as the units of analysis, either through choice or necessity. The design of these areas can greatly influence any observed spatial relationships or patterns-an effect known as the modifiable areal unit problem. In this paper we identify the phenomena and processes which are typically measured in environment and health studies and present a conceptualisation for their representation as data objects in spatial analysis. We discuss the circumstances under which we find ourselves using areas for representation and outline the application of zone design techniques for the creation of such areas in environment and health studies. An empirical study of the relationship between deprivation and limiting long-term illness in the former county of Avon, UK, is employed to demonstrate the potential usefulness of zone design techniques for creating zones with stable estimates and for exploring the sensitivity of relationships to changes in the zoning system. In particular, we illustrate the inappropriateness of the 1991 Census enumeration district and ward zoning systems for such an analysis and conclude that automatically designed aggregations may be a more appropriate basis for analysis than any pre-existing zoning system. PMID- 15820584 TI - The index of multiple deprivation 2000 access domain: a useful indicator for public health? AB - The access domain of the UK index of multiple deprivation (IMD) 2000 was designed to identify populations in small areas with poor geographical access to certain local key services. The measure is a composite of straight line distances to post offices, large food shops, primary schools and general practice surgeries for population sub-groups. Using the region of East Anglia as a case study area, this research evaluated the utility of the IMD2000 as an indicator of access to primary care. IMD2000 access scores for electoral wards were compared with a range of more detailed indicators of travel times and bus availability for visiting a general practitioner generated in a geographical information system (GIS). A range of easy-to-calculate surrogate variables was developed and tested as possible candidates to improve the explanatory power of the IMD2000 access score. The access domain was negatively correlated with the other five deprivation domains that comprise the overall index, suggesting that access should not be combined with the other measures of deprivation into a composite single score. The access domain was also found to predict access to primary care only with moderate accuracy. Two additional indicators of accessibility calculated in a GIS (road kilometres per thousand population and the presence of a major road in each ward) were found to add slightly to the power of the index. The predictive power of the index was best in urban areas, although it is in rural areas that access to primary care is a more important public health issue. The IMD2000 should be therefore used with caution as a measure of health service accessibility in rural areas. PMID- 15820585 TI - Selective migration, health and deprivation: a longitudinal analysis. AB - Population migration is a major determinant of an area's age-sex structure and socio-economic characteristics. The suggestion that migration can contribute to an increase or decrease in place-specific rates of illness is not new. However, differences in health status between small geographical locations that may be affected by the inter-relationships between health, area-based deprivation and migration are under-researched. Using the Office for National Statistics (ONS) England and Wales Longitudinal Study (LS) 1971-1991, this research tracks individuals to identify any systematic sorting of people that has contributed to the area-level relationships between health (limiting long-term illness and mortality) and deprivation (Carstairs quintiles). The results demonstrate that among the young, migrants are generally healthier than non-migrants. Migrants who move from more to less deprived locations are healthier than migrants who move from less to more deprived locations. Within less deprived areas migrants are healthier than non-migrants but within deprived areas migrants are less healthy than non-migrants. Over the 20 year period, the largest absolute flow is by relatively healthy migrants moving away from more deprived areas towards less deprived areas. The effect is to raise ill-health and mortality rates in the origins and lower them in the destinations. This is reinforced by a significant group of people in poor health who move from less to more deprived locations. In contrast, a small group of unhealthy people moved away from more deprived into less deprived areas. These countercurrents of less healthy people have a slight ameliorating effect on the health-deprivation relationship. Whilst health deprivation relationships are more marked for migrants there are also health (dis ) benefits for non-migrants if their location becomes relatively more or less deprived over time. Overall we found that between 1971 and 1991, inequalities in health increased between the least and most deprived areas, compared with the health-deprivation relationship which would have existed if peoples' locations and deprivation patterns had stayed geographically constant. Migration, rather than changes in the deprivation of the area that non-migrants live in, accounts for the large majority of change. PMID- 15820586 TI - Housing improvement and self-reported mental distress among council estate residents. AB - This paper is concerned with how housing improvements instigated either publicly or privately influence the degree of psychological stress reported by council estate residents in South Manchester. Stress is measured on the GHQ12 scale containing standard symptomatic items. Potential sources of variation in this indicator are analysed within a geographical setting where repeated samples of residents were drawn from two adjacent suburban council housing estates before and after the implementation of a single regeneration budget (SRB) housing initiative in late 1999. The residents of one of these estates (Wythenshawe) were targeted by this funding while those in the other (Mersey Bank) were not. The latter, therefore, serve as a control for the effects of the enhanced incidence of housing improvement activity promoted by this SRB. Regression analyses revealed that stress was raised significantly among the SRB residents perhaps on account of the additional environmental nuisance they encountered. The experience of stress among all residents, however, was dominated by measures of personal psychosocial risk and it is argued that future regeneration initiatives should address the manifestation of these risks in the effort to achieve better mental health. PMID- 15820587 TI - The impacts of dwelling conditions on older persons' psychological well-being in Hong Kong: the mediating role of residential satisfaction. AB - About 11% of Hong Kong's population of 7 million people are aged 65 and over and many of them live in old urban areas. Many of these areas have been subjected to urban redevelopment and some of the residents have been relocated to newer estates in peripheral new towns. Previous studies have focused on the challenges the urban environment has placed on older persons in terms of capability to cope with the demands that the environment places upon them. This paper suggests that dwelling conditions can act as stressors and become contributing factors that impact on older persons' residential satisfaction and psychological well-being (subjective well-being). This study examines the role of residential satisfaction (satisfaction with dwelling unit, estate and district) in mediating the effects of dwelling conditions (interior environment and exterior environment) on psychological well-being. A sample of older persons was recruited from a sampling frame of 16 urban sub-areas located in old urban areas and new towns. 518 older persons (224 males, 294 females) aged 60 and over were interviewed and the findings indicated that residential satisfaction was determined by assessment of both the interior environment and the exterior environment, although these were appraised differently. The interior environment had a greater impact on residential satisfaction than the exterior environment. It appeared that environmental dwelling conditions mainly affected older persons' psychological well-being indirectly and, hence, probably influenced their opportunities for successful ageing. However, subsequent tests revealed that dwelling conditions had no direct impact on psychological well-being. In light of these findings, it is proposed that the role of environmental factors and their relation to older persons' psychological well-being depends on the extent to which a person's expectations of residential satisfaction are met. Some implications of these findings for local housing and social care policy are discussed. PMID- 15820588 TI - Who you know, where you live: social capital, neighbourhood and health. AB - This article examines the degree to which relationships between social capital and health are embedded in local geographical contexts and influenced by demographic factors, socio-economic status, health behaviours and coping skills. Using data from a telephone survey of a random sample of adults (N=1504 respondents, response rate=60%), the article determines if relationships between involvement in voluntary associations and various measures of individual health are associated with neighbourhood of residence in the mid-sized city of Hamilton, Canada. Associational involvement and overweight status (assessed by body-mass score) were weakly but significantly related after controlling for the other variables; involvement had relationships with self-rated health and emotional distress before but not after controlling for socio-economic status, health behaviours and coping skills. Relationships between neighbourhood of residence and two health outcomes, self-rated health and overweight status, were statistically significant before and after controlling for the other characteristics of respondents; neighbourhood of residence was not a significant predictor of number of chronic conditions and emotional distress in multivariate models. The neighbourhood and associational involvement relationships with health were not dependent upon one another, suggesting that neighbourhood of residence did not help to explain the positive health effects of this particular measure of social capital. PMID- 15820589 TI - Family, friend or foe? Critical reflections on the relevance and role of social capital in health promotion and community development. AB - Social capital has been the focus of considerable academic and policy interest in recent years. Despite this interest, the concept remains undertheorized: there is an urgent need for a critical engagement with this literature that goes beyond summary. This paper lays a foundation for a critical dialogue between social capital and health promotion, by examining problematics in the conceptualization and practice of social capital building and linking these to models of community development, a cornerstone health promotion strategy. In so doing, the paper contributes to the existing literature by providing a theoretical exposition and critique of various threads in social capital discourse, and linking these threads explicitly to community development practice. Distinctions between communitarian, institutional and critical approaches to social capital are elaborated, and the relationships between these three approaches and three models of community development-social planning, locality development, and social action are discussed. The existing social capital literature is then critically examined in relation to three key themes common to both literatures: community integration, public participation, and power relations. This examination suggests that social capital cannot be conceived in isolation from economic and political structures, since social connections are contingent on, and structured by, access to material resources. This runs counter to many current policy discourses, which focus on the importance of connection and cohesion without addressing fundamental inequities in access to resources. This paper posits that approaches to community development and social capital should emphasise the importance of a conscious concern with social justice. A construction of social capital which explicitly endorses the importance of transformative social engagement, while at the same time recognising the potential negative consequences of social capital development, could help community organizers build communities in ways that truly promote health. PMID- 15820590 TI - Is the urban excess in lung cancer in Scotland explained by patterns of smoking? AB - Numerous studies have shown that lung cancer rates are higher in urban than rural areas, controlling for differences in age and sex profiles. One explanation is that smoking rates are higher in urban areas, although it is not clear whether the variations in smoking behaviour fully account for the observed urban/rural gradient in lung cancer incidence. Indeed, some studies have demonstrated an excess of cases in urban areas, even controlling for smoking behaviour. However, previous studies have been hampered by the lack of small-area smoking estimates which are required if urban/rural variations are to be examined reliably. This paper considers whether there is an urban excess in lung cancer incidence in Scotland, a country with particularly high rates of the disease, for the period 1988-1991. First, we examine whether an urban excess exists in Scotland using Poisson probabilities and a cluster detection technique. Second, regression analysis was then used to test whether any urban excess in lung cancer incidence remained once smoking behaviour was controlled for, using smoking estimates calculated for small areas throughout Scotland. The results demonstrate that the rates of lung cancer were higher in urban areas and that all the significant clusters of cases of lung cancer were located in the large urban centres of Scotland. Smoking behaviour did account for much of this urban excess in lung cancer, although it did not explain the entire effect. These results suggest that there are urban effects that influence the incidence of lung cancer that are not explained entirely by smoking behaviour. Possible explanations include the variations in exposure to air pollution, occupational differences and the legacy of selective migration between urban and rural areas. PMID- 15820591 TI - Particulate air pollution, social confounders, and mortality in small areas of an industrial city. AB - Scientists and policymakers have shown growing interest in the health effects of chronic air pollution exposure. In this study, we use geostatistical techniques in combination with small-area data to address a central research question: "Does chronic exposure to particulate air pollution significantly associate with mortality when the effects of other social, demographic, and lifestyle confounders are taken into account?" Our analysis relies on age-standardized mortality ratios for census tracts (CTs) of Hamilton (average population of 3419 persons), social and demographic data from the 1991 Census of Canada, smoking variables extracted from secondary surveys, and total suspended particulate (TSP) data from 23 monitoring stations operated by the Ministry of the Environment. Air pollution data are interpolated with a geostatistical procedure known as "kriging". This method translates fixed-site pollution monitoring observations into a continuous surface, which was overlaid onto the population-weighted centroids of the CTs. Our results show substantively large and statistically significant health effects for women and men. Evaluated over the inter-quartile range of the data, we found the relative risk of premature mortality for TSP exposure to be 1.19 (95% CI: 1.13-1.26) for women and 1.30 (95% CI: 1.24-1.37) for men. We also tested associations with cardio-respiratory and cancer mortality. We found positive, significant associations between particulate exposure and these causes of death in most models. Inclusion of socioeconomic, demographic, and lifestyle reduced but did not eliminate the health effects of exposure to particulate air pollution. Overall our results suggest that intra urban variations in particulate air pollution significantly associate with premature, all-cause, cardio-respiratory, and cancer mortality in small areas of Hamilton. PMID- 15820592 TI - Geographical inequalities of mortality by income in two developed island countries: a cross-national comparison of Britain and Japan. AB - In this paper we examine the ecological relations between household income distribution and age-grouped mortality in Britain and Japan. Comparable datasets were prepared in terms of age intervals of mortality, household income intervals and geographical units for years around 1990. Then we conducted a series of regression analyses to associate absolute and relative income indices with age and sex-specific standardized mortality ratios (SMRs). The results are as follows: (1) In Britain mortality is lower where inequalities in income are lower, while in Japan there is no obvious relationship. It is, however, apparent that-just as in the case of the USA and Canada-Britain and Japan appear to merge and appear part of a greater pattern when considered as a series of city regions. Thus an overall global relationship between income inequality and mortality may exist. To assess such global relationship, further studies using cross-national regional datasets covering a wide rage of rich nations are desirable. (2) Income mortality relations are consistent among different age-sex groups in Britain, but there are substantial differences in the relationships as revealed between different demographic groups in Japan. In particular, while absolute income levels are correlated negatively with mortality of working-age men in both countries, mortality of elderly people in Japan is higher where absolute income is higher. This indicates the different historical contexts to the health divides these two different geographical contexts, but further consideration of a more historically nuanced understanding of income-mortality relations is required. PMID- 15820593 TI - Urban/rural inequalities in suicide in Scotland, 1981-1999. AB - Although suicide accounts for a small percentage of deaths in Scotland (1.4% in 1999), it has been steadily increasing over the last two decades. In the US, Australia, England and Wales the greatest rises in suicide for this time period, occurred in rural areas. This study describes the pattern and magnitude of urban/rural variation in suicide in Scotland, examines methods of suicide within differing geographies and looks at trends in suicides over time. Scotland is split into four rurality types. Suicide data for all areas of Scotland (apart from Grampian which underwent changes in postcode sector boundaries in 1996) are investigated using Standardised Mortality Ratios (SMRs) and multilevel Poisson modelling, adjusting for age, sex and deprivation. SMRs for 1981-85, 1989-93 and 1995-99 are created across the four geographies, using the populations of Scotland in 1983, 1991 and 1997 as the standard populations (SMR=100). The highest rates in 1995-99 are seen in "remote rural" areas, SMR=125 (95% confidence interval 107-146). Models adjusted for age and deprivation show significantly greater risk of male suicide in remote rural areas relative to urban areas and significantly lower risk of female suicide in accessible rural areas. The method of suicide varies across ruralities for both males and females. The study considers how the relationship between suicides and rurality varies over time and how methods of suicide vary across different ruralities. The steepest rises in suicide amongst men, adjusting for age and deprivation, were seen to occur in accessible rural areas, however highest rates remain in remote rural areas. PMID- 15820594 TI - [What about sentinel node practice in early-staged cutaneous melanoma in France in 2003?]. AB - AIM: The aim of this study was to establish the status of sentinel lymph node (SLN) biopsy procedure in cutaneous melanoma in France in 2002. MATERIAL AND METHODS: This study was based upon the statistics of the main French melanoma centers. A short questionnary was sent to Head Physician by email. The authors asked for the global attitude as far as SLN was concerned, number of cutaneous melanoma diagnosed during year 2002 and of SLN procedures performed, critters of inclusion and postoperative management in each case. Abstension could be argued in a free item. Answers were sent back by email. RESULTS: The authors collected 22 answers coming from overall territory; 64% performed SLN procedure (14 centers), 36% applied "wait and watch" policy. Staffs performing SLN diagnosed a mean of 101 (8-400) melanoma and biopsied a mean of 21 (0-53) sentinel nodes. The others diagnosed a mean of 151 (15-250) melanoma. Patients were enrolled for Breslow thickness upper to 1.5 mm in 71%, to 1 mm in 29%. Ulceration was a critter of inclusion in 93% (21 staffs), 100% enrolled patients whose tumor presented signs of regression. SLN was performed for primary sites located overall body in 71%, only in limbs and trunk in 29%. Positive node lead to regional lymph node clearance, then observation or interferon protocol. Negative node lead to "wait and watch policy" in 14%, different interferon protocols according to Breslow thickness in 86%. CONCLUSION: SLN procedure is not homogenous in France. France is divided as far as SLN is concerned. If 64% are performing SLN, more than 50% of the new melanoma are not included in the trial. PMID- 15820595 TI - [Analysis of lymph mode recurrence in patients with melanoma and negative sentinel lymph node negative. Retrospective study at the Hospital Saint-Louis, Paris]. AB - Melanoma is a malignant tumor, with dominant lymphatic extension. Sentinel lymph node is the first lymph node touched by melanoma. Our retrospective and monocentric study is about 87 patients, between July 1999 and July 2003. The inclusion criteria were malignant melanoma with Breslow level superior or equal 1.5 mm, and/or Clark level superior or equal IV, and/or ulcerated, and/or in regression. Sentinel lymph node has been negative on histological analysis in 75 patients (86.2%). About these 75 patients, we found five metastatic lymph node recurrence (6.66%) in a short notice (median 10.2 months). For the five patients with recurrence, the original slides and tissue blocks were available for reexamination. Then, we found micrometastasis in two patients (40% of occult metastasis). Our rate of lymph node recurrence in patients with sentinel lymph node negative is about 6.66%. Our analysis make us believe that early recurrence are essentially linked to histological analysis limits, and maybe to skip metastasis existence. PMID- 15820596 TI - [Is sentinel lymph node procedure in early-staged cutaneous melanoma really safe?]. AB - AIM: The aim of this study was to analyze outcome in patients enrolled in sentinel node biopsy procedure in early-staged cutaneous melanoma. MATERIAL AND METHODS: Therefore a prospective study was conducted enrolling patients presenting with early-staged cutaneous melanoma. Our study focused on age and sex, duration from diagnosis to treatment, duration of hospitalization, dressing care and work inability in current follows. Duration from scintigraphy to surgery was analyzed and compared to sensibility of the procedure. What is more we observed rate and kind of complications and economical consequences, increasing duration of dressing care and work inability. The authors aimed at evaluating costs of SLN procedure including hospitalization, lymphoscintigraphy, general anaesthesia, costs of dressings, inability and overcosts of complications. RESULTS: Forty-five patients were enrolled in our study (sex-ratio 1/2) mean aged 60 years old. Duration from diagnosis to treatment was mean 36 days. Sensibility of the procedure was excellent in trunk and limbs cases, lower in head and neck. In current cases patients were mean hospitalized three days, underwent 20 days of dressings and work inability depended on further interferon treatment. Complications occurred in 25% as seroma or local infections requiring antibiotherapy. Duration to healing was then 45 days increasing inability. Global costs of SLN procedure were significantly higher than previous wait and watch policy. CONCLUSION: SLN biopsy is an expensive and invasive procedure with a high rate of complications. It defers melanoma treatment, only way to gain survival. PMID- 15820597 TI - [Reduction mammaplasty using superior pedicle in macromastia]. AB - SUBJECT: The authors present technical details, complications, morphologic and aesthetic results of 26 breast reduction mammaplasty for macromastia (breast reduction more than 1000 g) showing advantages and reliability of technique. MATERIALS AND METHODS: From January 2000 to December 2001, 223 patients underwent bilateral reduction mammaplasty with superior-based pedicled dermo-glandular flap. In 26 of them the weight of removed mammary tissue was over 1000 g in each breast. These 26 cases were evaluated, and the criteria adopted to analyse the results was morphologic and aesthetic evaluation of patient herself (very good, good, acceptable, unacceptable). RESULTS: Mean follow-up for all patients was 15 months. Twenty-six patients (mean age 33.2 years) underwent an average weight of 1131 g (930/2200 g) removed per breast. The following complications were observed: 1 Nipple Areolar Complex ischemia without necrosis; three infections (abscess); four delayed wound closure. The patient subjective evaluation of result was: "very good" in 19 cases (73%); "good" in 5 cases (19.2%) and "acceptable" in the others two cases (7.8%). No case was evaluated "unacceptable". CONCLUSION: Superior dermoglandular pedicle mammaplasty represent a very good and reliable solution for the treatment of macromastia, giving satisfactory cosmetics results with good nipple viability without necrosis. This technique is actually our first choice in the management of macromastia. PMID- 15820598 TI - [Breast carcinoma diagnosed from surgical specimens. Retrospective study on three years]. AB - In the wake of three consecutive cases of microscopical examination of resection specimens following breast reduction revealing an adenocarcinoma, we wanted to point out the interest of a complete preoperative senological examination including mammography and postoperative anatomopathological examination. A retrospective study concerning 837 patients over a three-year period was conducted. We found seven patients (0.83%) with malignant breast cancer diagnosed on anatomopathological examination, which is comparable to the incidence found in literature. Of these seven cases there were four ductal adenocarcinomas (0.47%), all of them in situ (DCIS), and three lobular adenocarcinomas (0.36%) of which one invasive (ILA), one in situ (LCIS) and one mixed. The majority was aggressive, multifocal and bilateral. Treatment consisted of mastectomy with or without adjuvant therapy with curative intent in five out of seven cases, and this within two months after a esthetic surgery. In our opinion this shows that breast reduction can help in tracking down breast cancer and underlines the need for systematic and meticulous microscopic examination of resection specimens after breast reduction. PMID- 15820599 TI - [Surgical site infection surveillance in breast implants surgery]. AB - The results of long-term follow-up of surgical site infection (SSI) after aesthetical breast surgery are reported. 205 consecutive patients operated from 1/2000 to 3/2002 were followed for at least one year. Postoperative incidents were observed in 26 (12.7%) patients, including SSI in six (2.9%) patients. After surgery, the mean time to SSI was 113 days, with only two cases in the first postoperative month. In all SSI cases, no antibiotic prophylaxis had been given. The causative role of infectious agents in breast implant capsule occurrence is under investigation. PMID- 15820600 TI - [Poland'syndrome and hand's malformations: about a clinic series of 37 patients]. AB - INTRODUCTION: Poland's syndrome is a rare malformation which associates thoracic anomalies and anomalies of homolateral upper end. We wish to know the frequency of hand's malformations in this syndrome in our clinical experience. MATERIAL AND METHODS: We have revised 37 patients who were seen initially for a thoracomammary anomaly. This clinical series from plastic surgery service of Toulouse has been revised to know the importance of hand's malformations. RESULTS: Hand's malformations in Poland's syndrome are rare in your study, they touch only 12% patients. We find only 4 malformations in 33 patients, four were lost. They were only females, we find three brachymesophalangies and a major form. DISCUSSION: Hand's malformations in Poland's syndrome are less frequent than classically. There is no parallelism between gravity of thoracic malformation and that one of upper end. In this series, we find only one case with syndactyly; originally, Poland's syndrome was named << Poland's syndactyly >>. Finally, we think that we can talk about Poland's syndrome without anomaly of homolateral upper end, the major element is musculary agenesia of sternocostal pectoralis major. The search of homolateral upper end has to be systematic in front of suspicious of Poland's syndrome. PMID- 15820601 TI - [The abdominal flap-graft for coverage of soft tissue defects of the dorsum of hand and fingers in burn patients. Six cases reports]. AB - We report six cases of abdominal flap-graft used to cover post-burn soft tissue defects of hands. The technique used was first described by Colson and is applied in our study to an abdominal donor site. Debulking of the flap at the time of elevation resulted in a skin quality as good as a full thickness graft without jeopardizing flap vitality and reliability. Digit mobilization was possible after separation. Long term functional results were found to be satisfactory, specially for the hand. For the fingers it is mostly a digit-rescue surgery. The application of this technique remains sparse. Still, the abdominal flap-graft is often useful in cases of burn patients because of the poor local conditions, as well as the magnitude and nature of injury, which limit the coverage options. PMID- 15820602 TI - [Joint denervations in hand surgery: a plea]. AB - Since the original description of wrist denervation, the principle of joint denervation in hand surgery has been extended to other joints like the first carpometacarpal, interphalangeal and metacarpophalangeal joints. At the wrist level as at the digital level, the results are comparable to those obtained with other surgical alternatives. In case of unsatisfactory results, a more traditional intervention remains always possible. PMID- 15820603 TI - [Full thickness skin graft of glabellar skin for the repair of nasal tip defect]. AB - The use of glabellar skin as full thickness skin graft is a good alternative to the common techniques, which are essentially represented by local flaps for repair of the nasal tip defects. Three cases out of 25 patients are reported. This technique provides no dyschromia unlike skin graft or composite graft commonly used. The glabellar scar remains remarkably discrete on a patient with previous glabellar wrinkles. The limits of this technique are represented by the surface of the donor skin its use on young patients without glabellar wrinkles. PMID- 15820604 TI - [Mammoplasty as preparatory step to elbow arthroplasty for complete ankylosis of the elbow: a case report]. AB - A 45-year old polytraumatized female patient presented with an ankylosed elbow two years after a traffic accident. A skin graft adherent to the proximal radius was painful and precluded any attempt at total elbow arthroplasty. Local muscular, cutaneous and vascular sequelae contraindicated the usual local regional flaps or made risky elbow coverage with free flaps. The patient's breast hypertrophy, an obstacle to a random-pattern flap on the torso, offered enough tissue to cover the injured elbow with an axial-pattern flap of the left breast lower-outer quadrant. Six months later the patient had a total elbow arthroplasty with pain suppression and improvement of the range of motion. PMID- 15820605 TI - [Galactorrhea after mammary plastic surgery]. AB - Galactorrhoea is a complication rarely observed after mammary plastic surgery. Our experience in the domain extends to three clinical cases - two after prosthetic insertion and one after breast reduction - wich will be presented here. The origin of this complication is uncertain. Nevertheless, it is likely to be multifocal, as surgery alone is not the only cause. Postsurgical galactorrhoea often follows a benign course culminating in spontaneous resolution. However, it may reveal the presence of o prolactin secreting adenoma, as was the case with one of our patients. A detailed history, exploring antecedent factors, is an essential step in guiding subsequent management. When faced with postsurgical galactorrhoea, serum prolactin levels should be measured. If serum prolactin levels exceed 150 ng/ml further investigation by way of an MRI of the sella turcica is advisable to rule out pituitary adenoma. Depending on symptom severity, treatment may be medical with the prescription of dopaminergic agonists, and/or surgical with drainage or removal of prostheses. Increased awareness of galactorrhea as a possible complication of plastic surgery to the breast will improve management. PMID- 15820606 TI - Dietary fatty acids intake: possible role in cognitive decline and dementia. AB - There is a recent increase in the level of interest in the possible role of dietary fatty acids in age-related cognitive decline, and cognitive impairment of both degenerative (Alzheimer's disease, AD) or vascular origin. At present, several studies suggested that an increase of saturated fatty acids (SFA) could have negative effects on cognitive functions. Furthermore, a clear reduction of risk of cognitive decline has been found in a population sample with a high intake of polyunsaturated fatty acids (PUFA) and monounsaturated fatty acids (MUFA). These findings were confirmed by studies in which high intakes of n-6 PUFA, n-3 PUFA, MUFA, and weekly fish consumption, providing large amount of n-3 PUFA, appear to be protective against the risk of AD. In our elderly population from Southern Italy, elevated unsaturated fatty acids intake (MUFA and PUFA), high levels of antioxidant compounds, and very low SFA intake could act synergistically in improving cognitive performance. Epidemiological studies on the association between diet and cognitive decline suggested a possible role of fatty acids intake in maintaining adequate cognitive functioning and possibly in preventing or delaying the onset of dementia, both of degenerative or vascular origin. Appropriate dietary measures or supplementation with specific micro- and macronutrients might open new ways for the prevention and management of cognitive decline and dementia. PMID- 15820608 TI - Age-related changes in sleep and memory: commonalities and interrelationships. AB - Aging is accompanied by changes on many different levels, including sleep and memory processes. In light of recent findings, which underline the importance of sleep for memory consolidation in young adults, the relationship between sleep and memory in older adults is of particular interest. This review takes a closer look at commonalities and interrelationships of the two aging trajectories in sleep and memory. A link between these two research domains on aging will be proposed and discussed on the basis of recent findings within the cognitive neuroscience of sleep. PMID- 15820607 TI - Changes in renal hemodynamics and structure in the aging kidney; sexual dimorphism and the nitric oxide system. AB - With advancing age the kidney shows both functional declines (falls in GFR) and development of structural damage. In most individuals this occurs slowly and does not lead to severe renal impairment unless additional insults are superimposed. There is a pronounced sexual dimorphism with females protected, due both to beneficial effects of the estrogens and damaging effects of androgens, some of which act directly on the glomerular mesangial cell to regulate growth and extracellular matrix production. Nitric oxide is a major factor in regulation of vascular tone and growth and becomes deficient with advancing age, as endothelial dysfunction develops. Although the abundance of the substrate, L-arginine, is well maintained during aging, there are increases in the concentration of circulating endogenous nitric oxide synthase (nNOS) inhibitors, which will contribute, to the endothelial dysfunction. There is a clear sexual dimorphism in the NO system, with pre-menopausal females producing more NO than men. Within the kidney, declines in the abundance and activity of the neuronal form of the nitric oxide synthase (nNOS) correlate with development of disease. In the male rat where injury and dysfunction occurs, nNOS abundance declines markedly, whereas in the protected female, renal nNOS abundance is maintained. Taken together, it is likely that age-dependent declines in NO generation contribute to age-dependent kidney damage. PMID- 15820609 TI - Defective cholesterol efflux in Werner syndrome fibroblasts and its phenotypic correction by Cdc42, a RhoGTPase. AB - Werner syndrome (WS) is characterized by the early onset of senescent phenotypes including premature atherosclerotic cardiovascular diseases, although the underlying molecular mechanism for atherosclerosis has not been fully understood yet. Cholesterol efflux from the cells is the initial step of reverse cholesterol transport, a major protective system against atherosclerosis. The aim of the present study was to determine whether this crucial step may be altered in WS. We examined intracellular lipid transport and cholesterol efflux and the expression levels of its related molecules in skin fibroblasts obtained from patients with WS. Cholesterol efflux was markedly reduced in the WS fibroblasts in association with increased cellular cholesterol. Fluorescent recovery after photobleaching (FRAP) technique revealed that intracellular lipid transport around Golgi apparatus was markedly reduced when using a C6-NBD-Ceramide as a tracer. Cdc42 protein and its GTP-bound form were markedly reduced in the WS fibroblasts. The complementation of wild-type Cdc42 corrected cholesterol efflux, intracellular lipid transport, and cellular cholesterol levels in the WS fibroblasts. These data indicated that the reduced expression of Cdc42 may be responsible for the abnormal lipid transport, which in turn might be related to the cardiovascular manifestations in WS. PMID- 15820610 TI - Administration of estradiol and progesterone modulate the activities of antioxidant enzyme and aminotransferases in naturally menopausal rats. AB - In aging tissues the oxidative stress increases due to decreased activity of antioxidant enzymes and proteolysis increases due to decreased activity of aminotransferases, which can be modified by hormonal replacement therapy (HRT). The aim of the present study was to determine the effect of HRT on the activities of an antioxidant enzyme superoxide dismutase (SOD) and aminotransferases like alanine aminotransferase (Ala-AT) and aspartate aminotransferase in different age groups (12, 18 and 24 months) of naturally menopausal rats. The rats were given the subcutaneous injection of 17beta-estradiol, progesterone and combination of estradiol and progesterone for 1 month. The activity of SOD, Ala-AT and Asp-AT was measured in the brain (cerebral hemisphere, CH), heart, liver, kidney and uterus. The activity of SOD decreased with age in all the tissues taken particularly in liver. After HRT the enzyme activities were increased as compared to age-matched controls in all the tissues of aging rats. The activities of transaminases (Ala-AT and Asp-AT) showed a decrease with age in all the tissues and administration of estradiol and combination of estradiol and progesterone further decreased both the aminotransferases. Our study elucidates that increased activity of SOD contributes in protection of cells from oxygen toxicity by catalyzing the dismutation of free radicals in tissues. Furthermore, the HRT probably decreases gluconeogenesis and proteolysis by decreasing the activities of Ala-AT and Asp-AT in aging rat tissues. PMID- 15820611 TI - Hyperglycemia, impaired glucose tolerance and elevated glycated hemoglobin levels in a long-lived mouse stock. AB - We have previously demonstrated that two wild-derived stocks of mice, Idaho and Majuro, are significantly longer-lived than mice of a control stock (DC) generated as a four-way cross of commonly used laboratory strains of mice. This study provides independent confirmation of this earlier finding, as well as examining serum glucose, insulin, leptin, glycated hemoglobin (GHb), cataract severity, and glucose tolerance levels in each of the stocks. Both the mean (+20%) and maximum (+13%) life span of the Idaho mice were significantly increased relative to the DC stock, while in the Majuro mice only maximum (+15%) life span was significantly increased. In addition, Majuro mice were hyperglycemic in both the fed and fasted states compared both to laboratory derived and Idaho stocks, had significantly elevated GHb levels and cataract scores, and were glucose intolerant although serum insulin levels did not differ between stocks. Body weight and body mass index (BMI)-corrected leptin levels were also dramatically (1.5-3-fold) higher in the Majuro mice. The longevity of Id mice was not accompanied by changes in serum glucose and insulin levels, or glucose tolerance compared to DC controls, although GHb levels were significantly lower in the Idaho mice. Taken together, these findings suggest that neither a reduction of blood glucose levels nor an increase in glucose tolerance is necessary for life span extension in mice. PMID- 15820612 TI - Expression of pulmonary VEGF family declines with age and is further down regulated in lipopolysaccharide (LPS)-induced lung injury. AB - Vascular endothelial growth factor (VEGF) is a survival factor in endothelial cells and a promoter of angiogenesis that reportedly plays a pivotal role protecting against injury. In aged humans and animals, lung injuries are generally more serious and cause higher mortality. We thus hypothesized that the expression of VEGF and its related molecules in the lung declines with age. In this study, we first examined the expression of VEGF family (VEGF-A, -B, -C and D), VEGF-A isoforms (VEGF120, 164, 188), and VEGF-specific receptors (VEGFR-1: Flt-1; VEGFR-2: Flk-1 and VEGFR-3: Flt-4) by quantitative RT-PCR in lungs from young and old mice. Expression of all these except for VEGF-D was significantly lower in old mice than in young mice. We then subjected young and old mice to lipopolysaccaride (LPS)-induced lung injury. Old animals demonstrated poor survival and prolonged lung inflammation when compared with young counterparts. At 24 and 72 h after intratracheal LPS administration, expression of the examined factors was down-regulated in the lungs irrespective of age. In conclusion, pulmonary expression of the VEGF family and their receptors declines with age, and is further down-regulated in LPS-induced lung injury, although the mechanism of age- and/or injury-related down-regulation of VEGF remains unknown. PMID- 15820613 TI - Differential effects of alcohol upon gluconeogenesis from lactate in young and old hepatocytes. AB - The purpose of this study was to determine the effect of age upon hepatic gluconeogenesis (HGN) from lactate in the presence of various concentrations of alcohol from young (3 months) and old (24 months) male rats. After a 24-hour fast, livers were perfused with collagenase and the hepatocytes were isolated. Aliquots of the cell suspension were placed in Krebs-Henseleit buffer and incubated with lactate, [U-(14)C]lactate, and nine different concentrations of ethanol (EtOH) for 30 min. Dose-effect curves were generated for the determination of maximal and half-maximal alcohol-induced inhibition on gluconeogenesis. There were no significant differences in basal HGN (lactate only and no EtOH) between young and old hepatocytes, 86.9+/-6.3 nmol/mg protein/30 min. The addition of ethanol significantly reduced HGN from lactate in both groups. At the highest ethanol concentration (15 mM), the glucose production was inhibited more from old, 46.1+/-1.2 nmol/mg protein/30 min, compared to young hepatocytes, 56.0+/-1.6 nmol/mg protein/30 min. The greater age-related reduction in HGN was confirmed by the minimal glycogenolysis, and the concomitant decline in [U-(14)C]glucose production, lactate uptake, and [U-(14)C]lactate uptake. The results suggest that alcohol elicits a greater inhibition upon HGN from lactate in old compared to young liver cells. PMID- 15820614 TI - Effects of ageing and pharmacological hypothyroidism on pituitary-thyroid axis of Dutch-Miranda and Wistar rats. AB - To evaluate the ability of the aged rat pituitary to increase TSH secretion in response to major decreases in serum thyroid hormones, hypothyroidism was induced by methimazole in young and old, male and female, Dutch-Miranda and Wistar rats. Before MMI-treatment there were no differences in serum TSH of young and old rats, but serum T(4) was significantly decreased in aged rats from both genders and strains, while serum T(3) was significantly decreased in aged male rats from both strains, and in old Wistar females. MMI treatment significantly decreased serum T(4) and T(3) in all treated animals, and progressively increased serum TSH in both male and female rats, but the increase was significantly smaller in the elder rats. The pituitary TSH content was higher in Wistar than in Dutch-Miranda rats, of both genders, and was not significantly affected by age. MMI treatment decreased the pituitary TSH in both young and old Dutch-Miranda rats, but in the Wistar strain only the old females had a significant decrease. Our results show that the ability of the pituitary thyrotrophs to increase hormonal secretion in response to decreased levels of thyroid hormones is impaired in the old rat, even when the thyroid hormone levels are dramatically reduced. PMID- 15820615 TI - Unsaturated fatty acids intake and all-causes mortality: a 8.5-year follow-up of the Italian Longitudinal Study on Aging. AB - Recent evidence suggested a protective role of dietary monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) intakes against several chronic diseases and, therefore, an increased human longevity. After a median follow-up of 8.5 years, we investigated the possible role of MUFA, PUFA, and other selected food groups in protecting against all-causes mortality in a population-based, prospective study, conducted in one of the eight centers of the Italian Longitudinal Study on Aging (ILSA), Casamassima, Bari, Italy. Out of 704 elderly subjects (65-84 years), 278 nondemented persons agreed to participate at the first survey (1992-1993). During the follow-up, there were 91 deaths. A semi quantitative food frequency questionnaire evaluating macronutrient daily intakes were performed at the first survey. Higher MUFA intake was associated with an increase of survival (hazard ratio 0.81, 95% CI 0.66-0.99), a higher unsaturated fatty acids (UFA) to SFA ratio (hazard ratio 1.20, 95% CI 0.99-1.45) increased total mortality only marginally, while no effect about other selected food groups were found. In conclusion, in this prospective study on older nondemented subjects with a typical Mediterranean diet, a higher MUFA intake increased survival, while a higher UFA/SFA ratio increased total mortality, but only marginally. PMID- 15820616 TI - IL-6 gene variation is not associated with increased serum levels of IL-6, muscle, weakness, or frailty in older women. AB - Elevated levels of the inflammatory cytokine IL-6 are associated with the development of disability, frailty, and mortality in older adults. These outcomes are likely mediated through inflammatory activity that alters hormones, skeletal muscle, and the immune system. Polymorphic variants in the IL-6 gene influence IL 6 expression. We hypothesized that IL-6 alleles associate with increased serum of IL-6, decreased muscle strength, and frailty, and tested this in the Women's Health and Aging cohorts. We genotyped 463 participants age 70-79, and identified three common IL-6 haplotype blocks for the Caucasian (n=363) and African American (n=100) subsets. Using linear and logistic regression, and adjusting for age, BMI, race, and osteoarthritis, we identified no significant or clinically meaningful relationship between any single IL-6 single nucleotide polymorphism (SNP) or any IL-6 haplotype and serum IL-6 level, grip, knee, or hip strength, or frailty. Given that the promoter SNP (rs1800795) has been reported to influence IL-6 levels and health outcomes, we performed a similar association study in the In Chianti population (n=266) and confirmed lack of association. These results suggest that IL-6 gene variation may not be an important factor in the determination of elevated IL-6 levels and related phenotypes found in older women. PMID- 15820617 TI - Expression of mRNA for ROS-generating NADPH oxidases in the aging stomach. AB - Oxidative damage is thought to play a key role in the aging of various organ systems. In this study, we have therefore analyzed mRNA expression of ROS generating NADPH oxidases in the aging stomach. Gastric biopsies of hospitalized geriatric patients were analyzed for histology (Sidney classification), and real time PCR was used to quantify mRNA expression of the superoxide-generating NADPH oxidases NOX1, NOX2, and NOX5. We found that stomach biopsies of elderly patients expressed NOX5 and NOX2 mRNA, but not NOX1. The mRNA expression of NOX5 (a lymphocyte NADPH oxidase) neither depended on age nor on the results of the stomach histology. In contrast, mRNA expression of NOX2 (phagocyte NADPH oxidase) was a function of two variables. Increased NOX2 mRNA levels were observed in biopsies with signs of chronic inflammation (p=0.01). Interestingly, however, there was also an age-dependent increase in NOX2 mRNA levels (p=0.01). We conclude that in elderly patients the gastric mRNA expression of the ROS generating enzyme NOX2 increases as a function of age, possibly contributing to stomach aging and gastric vulnerability of the elderly. PMID- 15820618 TI - Distribution of the NMDA receptor NR3A subunit in the adult pig-tail macaque brain. AB - The NMDA subtype of glutamate receptors are heteromeric complexes comprised of multiple subunits encoded by at least seven different genes (NR1, NR2A-2D and NR3A-3B), and differential expression of these subunits alters the pharmacological and electrophysiological properties of NMDA receptors. NR3A is a recently identified unique modulatory subunit that decreases NMDA receptor current and calcium influx. In rodents, NR3A is developmentally expressed, displaying robust expression early in development that declines with age, reaching low levels in the adult brain. A distinct and highly selective pattern of expression is observed in the developing and mature rodent brain, suggesting that NR3A may play a very specific role in NMDA receptor-mediated processes. NR3A expression in other species, however, is unknown. Therefore, we examined the expression of NR3A mRNA and protein in the adult macaque brain. Our results indicate that NR3A mRNA is expressed throughout much of the adult primate brain, and at high levels in specific brain regions including the neocortex, substantia nigra par compacta and cerebellum, as well as select areas of the hippocampus, amygdala, thalamus and hypothalamus. Western blot analysis reflects that this protein is translated and expressed in multiple brain regions. In contrast to the rat mRNA, our results suggest that NR3A transcript is widely expressed in the adult primate brain. Particular enrichment in some brain areas may reflect brain region or circuit-specific functions for this NMDA receptor subunit. PMID- 15820619 TI - Vitamin E against oxidative damage caused by formaldehyde in frontal cortex and hippocampus: biochemical and histological studies. AB - Formaldehyde (FA) can cause severe central nervous system impairment. But, there are only a few studies about biochemical and histopathological changes of frontal cortex and hippocampal tissue caused by FA toxicity. The aim of our study was to investigate these changes occurring after chronic formaldehyde toxicity in frontal cortex and hippocampal tissues, and protective effect of Vitamin E (vit E) against oxidative damage. Eighteen rats were divided into three groups: (1) control, (2) treated with FA (FAt), and (3) treated with FA and vit E (FAt+vit E) groups. After the treatment, the animals were sacrificed and frontal cortex and hippocampal tissues were removed for biochemical and histopathological investigation. FA significantly increased tissue malondialdehyde (MDA) and protein carbonyl (PC) levels and also decreased superoxide dismutase (SOD) and catalase (CAT) enzyme activities in frontal cortex and hippocampal tissue compared to control. Vit E treatment decreased MDA and PC levels and prevented inhibition of SOD and CAT enzymes in the tissues. In the FAt group, the neurons of both tissues became extensively dark and degenerated with picnotic nuclei. The morphology of neurons in FAt+vit E group was protected well, but not as neurons of the control group. The number of neurons in frontal cortex and hippocampal tissue of FAt group was significantly less than both control and FAt+vit E groups. It was concluded that vit E treatment might be beneficial in preventing FA-induced oxidative frontal cortex and hippocampal tissue damage, therefore, shows potential for clinical use. PMID- 15820621 TI - Regional analysis of neurofilament protein immunoreactivity in the hamster's cortex. AB - The laminar distribution of several distinct populations of neurofilament protein containing neurons has been used as a criterion for the delineation of cortical areas in hamsters. SMI-32 is a monoclonal antibody that recognizes a non phosphorylated epitope on the medium- and high-molecular weight subunits of neurofilament proteins. As in carnivores and primates, SMI-32 immunoreactivity in the hamster neocortex was present in cell bodies, proximal dendrites and axons of some medium and large pyramidal neurons located in cortical layers III, V and VI. A small population of labeled multipolar cells was also found in layer IV. Neurofilament protein immunoreactive neurons were found throughout isocortical areas. Very few labeled cells were encountered in supplemental motor area, insular cortex, medial portion of associative visual cortex and in parietal association cortex. Our data indicate that SMI-32 immunoreactive cells can be efficiently used to trace boundaries between neocortical areas in the hamster's brain. The regional distribution SMI-32 immunoreactivity in the hamster cortex corresponds quite closely with cortical areas as defined by their cytoarchitecture and myeloarchitecture. The primary sensory cortical areas contain the most intense of SMI-32 immunoreactivity and are also those with the highest density of myelinated axons. Very low SMI-32 immunoreactivity was found in orbital, insular, perirhinal, cingulate and infralimbic cortices, which are also poor in myelinated axons. This supports the association between SMI-32 immunoreactivity and myelin contents. PMID- 15820620 TI - Nitroxidergic neurons in rat nucleus tractus solitarii express vesicular glutamate transporter 3. AB - Earlier we reported that glutamate transporter (VGLUT) 2 and neuronal nitric oxide synthase (nNOS) are colocalized in some fibers and are present in apposing fibers in the nucleus tractus solitarii (NTS). Those findings provided anatomical support for a hypothesized physiological link between glutamate and nitric oxide (NO.) in the NTS. Recently a third class of VGLUT, VGLUT3, was identified, but its distribution in NTS and its anatomical relationship with nNOS have not been shown. In this study we tested the hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS. We perfused rats and obtained brain stem sections and nodose ganglion sections for immunofluorescent staining analyzed by confocal microscopy. The NTS contained moderate VGLUT3 immunoreactivity (IR), with the intermediate, medial and interstitial subnuclei containing higher VGLUT3-IR than other subnuclei. Although all three forms of VGLUT were present in the NTS, VGLUT3-IR was not colocalized with either VGLUT1 IR or VGLUT2-IR in either processes or cells in the brain stem. Cells and processes containing both VGLUT3-IR and nNOS-IR were noted in all NTS subnuclei and in the nodose ganglion. Triple immunofluorescent staining revealed that cells double-labeled for nNOS-IR and VGLUT3-IR were all additionally labeled for neuronal nuclear antigen (NeuN), a neuronal marker. These findings support our hypothesis that neurons and fibers containing VGLUT3 lie in close proximity to those containing nNOS and that both proteins colocalize in some neurons and fibers in the NTS. PMID- 15820623 TI - Efferent connections of the "olfactostriatum": a specialized vomeronasal structure within the basal ganglia of snakes. AB - The olfactostriatum is a portion of the basal ganglia of snakes that receives substantial vomeronasal afferents through projections from the nucleus sphericus. In a preceding article, the olfactostriatum of garter snakes (Thamnophis sirtalis) was characterized on the basis of chemoarchitecture (distribution of serotonin, neuropeptide Y and tyrosine hydroxylase) and pattern of afferent connections [Martinez-Marcos, A., Ubeda-Banon, I., Lanuza, E., Halpern, M., 2005. Chemoarchitecture and afferent connections of the "olfactostriatum": a specialized vomeronasal structure within the basal ganglia of snakes. J. Chem. Neuroanat. 29, 49-69]. In the present study, its efferent connections have been investigated. The olfactostriatum projects to the main and accessory olfactory bulbs, lateral cortex, septal complex, ventral pallidum, external, ventral anterior and dorsolateral amygdalae, bed nucleus of the stria terminalis, preoptic area, lateral posterior hypothalamic nucleus, ventral tegmental area, substantia nigra and raphe nuclei. Tracer injections in the nucleus accumbens proper, a structure closely associated with the olfactostriatum, result in a similar pattern of efferent connections with the exception of those reaching the main and accessory olfactory bulbs, lateral cortex, external, ventral anterior and dorsolateral amygdalae and bed nucleus of the stria terminalis. These data, therefore, help to characterize the olfactostriatum, an apparently specialized area of the nucleus accumbens. Double labeling experiments after tracer injections in the nucleus sphericus and the lateral posterior hypothalamic nucleus demonstrate a pathway between these two structures through the olfactostriatum. Injections in the olfactostriatum and in the medial amygdala show parallel projections to the lateral posterior hypothalamic nucleus. Since this hypothalamic nucleus has been previously described as projecting to the hypoglossal nucleus, both, the medial amygdala and the olfactostriatum may mediate vomeronasal influence on tongue-flick behavior. PMID- 15820622 TI - Visually-induced NGFI-A protein expression in the calbindin-, parvalbumin- and nitric oxide synthase-neuronal populations of the rat superior colliculus. AB - The expression of the immediate early gene NGFI-A in the nervous system is induced by sensory stimulation and seems to be related to long-term synaptic plasticity. We have used double-labeling immunohistochemistry to identify calbindin (CB)(+), parvalbumin (PV)(+) and neuronal nitric oxide synthase (nNOS)(+) neurons that also expressed the protein encoded by this immediate early gene after light-exposure on in the superficial layers of the rat superior colliculus (sSC). The majority of the NGFI-A(+) cells were not double-labeled for the tested markers. In the stratum zonale+stratum griseum superficiale (SZ/SGS), only 17.8%, 8.0% and 12.1% of NGFI-A(+) cells were also labeled for CB, PV or nNOS, respectively. In the stratum opticum (SO), only 10.5% of the NGFI-A(+) cells were also CB(+). Furthermore, only a small subset of each population expressed the NGFI-A protein after light-exposure. In the SZ/SGS, 35.7% of the CB(+), 32.1% of the PV(+) and 26.6% of the nNOS(+) neurons also expressed the NGFI-A. In the SO, 31.7% of the CB(+) neurons also expressed the NGFI-A. The proportional distribution of the nNOS(+)/NGFI-A(+) neurons throughout the SZ/SGS layers showed a slight but significant rostro-caudal gradient. No significant difference was observed for the other markers, indicating homogeneous activation of these populations throughout the retinotopic map. Our results suggest that the visually-driven NGFI-A expression is not restricted to a specific population of the sSC and that visual processing in this structure, as assessed by the expression of this candidate-plasticity protein, involves the activation of subsets of ascending and non-ascending projection neurons. PMID- 15820624 TI - Neural correlates of intentional and incidental recognition of famous faces. AB - Event-related potentials (ERPs) were used to study the relationship between intentional and incidental recognition of famous faces. Intentional and incidental recognition were operationally defined as repeated presentations of targets and nontargets within a modified Sternberg task. These repetitions elicited temporally and topographically distinct ERP modulations. A repetition effect around 300 ms (ERE/N250r) and a preceding modulation did not differ between intentional and incidental recognition, whereas a following repetition effect (LRE/N400) around 500 ms showed differences between incidental and intentional recognition. These results show that during the first few hundred milliseconds intentional and incidental face recognition relate to similar processing, indicating that familiar faces are recognized even when their identification is not required. PMID- 15820625 TI - Neural substrates in judgment process while playing go: a comparison of amateurs with professionals. AB - A professional go player shows incomparable ability in judgment during go game. Positron emission tomography (PET) was used to investigate the neural substrates of professional go player's judgment process. Eight professional go players and six amateur players were instructed to think over silently in the opening-stage game (fuseki, territorial planning) problems and the life-or-death (tsume, checkmate judgment) problems presented on the monitor in front of them for 60 s of H2 15O PET scans and to state the answer afterwards. We found that in the territorial planning problems the parietal activation was equally observed in both groups with the additional prefrontal activation in the amateur group, and in the checkmate-decision problems the precuneus and cerebellum were activated in professionals while the premotor and parietooccipital cortices (visuospatial processing region) were extensively activated in amateurs. The comparison of the two groups showed stronger activations in the precuneus and cerebellum in the professionals in contrast to the premotor activation in amateurs during checkmate judgment. In addition, the cerebellum was remarkably activated in the higher ranking professional players. These findings suggested the cerebellum and precuneus play important roles in processing of accurate judgment by visual imagery and nonmotor learning memory processes in professional go players. PMID- 15820626 TI - Enhanced contrast sensitivity in auditory cortex as cats learn to discriminate sound frequencies. AB - To better understand the nature and time course for learning-induced cortical reorganization, we examined frequency-specific changes in auditory cortex as cats gradually improved at a difficult sound frequency discrimination task. Three adult cats were trained to discriminate between a tone pip at a fixed target frequency (S-) and a higher deviant frequency (S+). An adaptive training schedule led to an efficient estimate of the frequency discrimination threshold (FDT), which was used to track daily performance. Each cat was also implanted with an array of microwires in auditory cortex. Tone pips with different frequency and amplitude were used to map receptive fields. Onset responses were correlated with training time and the cat's ability to discriminate frequencies. Although lifetime of the neural implants varied among cats, each provided sufficient neural recording to relate at least 3 weeks of learning to response changes in the cortex. An improved FDT was associated with a differential decrease in response strength between the S- frequency and S+ frequencies. Response to the training frequencies gradually located in a local minimum compared to adjacent frequencies (p < 0.001, Cohen's d=0.50). Cortical changes were consistent with a theory of bimodal generalization that enhances stimulus classification by reducing similarity between reinforced and nonreinforced stimuli. Such a strategy may be especially appropriate during an early stage of learning to discriminate similar sounds and differ from later strategies required for fine discrimination. PMID- 15820627 TI - The late negative episodic memory effect: the effect of recapitulating study details at test. AB - An hypothesis concerning mnemonic function suggests that perceptual details of previously experienced episodes are retrieved from the cortices that initially processed that information during the encoding phase. Cycowicz et al. [Cycowicz, Y.M., Friedman, D. and Snodgrass, J.G., Remembering the color of objects: an ERP investigation of source memory, Cereb Cortex, 11 (2001) 322-334.] have interpreted the presence of a late negative episodic memory (EM) effect, maximal over parieto-occipital scalp, as a brain signature of the search for and/or retrieval/evaluation of the specific perceptual source-specifying attributes (i.e., color) of pictures in the visual cortical regions that were recruited during the encoding of that information. The present study assessed the validity of this hypothesis. Twelve participants studied pictures outlined in red or green and were subsequently tested with inclusion (i.e., item; old or new regardless of color) and exclusion (i.e., source; same color, different color/new judgments) tasks. In both, old pictures were presented either in the same color as at study or in the alternate color. A late negative, parieto-occipital EM effect was of much larger amplitude in the source compared to the item task. It was of similar magnitude to correctly recognized pictures whose colors were identical at study and test relative to those whose colors changed, and was not modulated by the success or failure of the source retrieval. These data run counter to the initial hypothesis that the late negative EM effect reflects the search for and/or retrieval of specific perceptual attributes such as color. Rather, the late negative EM effect may reflect the search for and/or retrieval/evaluation of more general source-specifying information in the cortical regions that initially processed the stimuli. PMID- 15820628 TI - A novel experimental paradigm for studying cognitive functions related to delayed response tasks in mice. AB - Rodents are the animals most commonly employed to model human cognitive functions, but serious problems arise from the non-selective use of behavioral paradigms that measure different processes in rodents than those found in humans. To avoid problems stemming from the use of different paradigms on humans and mice, a new experimental paradigm for mice was developed to study the cognitive functions involved in delayed response tasks. The experiments were conducted in an olfactory tubing maze using three successive delayed response tasks: an alternation task, a non-alternation task, and a reversal task. Mice had to discover the rule by themselves by choosing one of two identical odor cues presented simultaneously at the left and right sides of a testing chamber. The success criterion was set at 10, 8, 6, or 4 consecutive correct responses, with a maximum of 80 trials per task, as used in primates. In the delayed alternation task with the criterion of 10 or 8 consecutive successful trials, the rule was discovered but required many more than 80 trials for most of the mice. With a criterion of 6 or 4, the mice were successful but twice as many trials were necessary to reach the criterion of 6 as opposed to 4. In the delayed non alternation and reversal tasks, more than 80 trials were needed to figure out the new rule with the criterion of 10 or 8. All mice were successful with the criterion of 6 or 4. The results indicated that no matter what criterion was used, mice were able to discover the two rules on the three consecutive delayed response tasks, but they did so with more or less ease. This novel paradigm for mice should be useful in experiments on pharmacological treatments or for testing transgenic or gene-targeting mice to gain insight into the brain structures involved in this type of task. PMID- 15820629 TI - An event-related fMRI study of the neural networks underlying the encoding, maintenance, and retrieval phase in a delayed-match-to-sample task. AB - Memory loads exceeding the limited capacity of working memory (WM) have been shown to expand the prefrontal areas that participate in WM and have revealed substantial individual differences in performance. We used a delayed-match-to sample (DMS) task in an event-related fMRI study to map the full extent of the expanded regional activations associated with supracapacity loads. A 6-letter study array was compared to arrays of 1 and 3 letters. The task comprised separate encoding, retention, and retrieval fMRI epochs. A brain-wide spatial covariance analysis was applied to the data of all task epochs to identify patterns of correlated regional activations whose expression increased monotonically across 3 memory-load levels on a subject-by-subject basis. Such load-related activation patterns were in all task phases. Of greatest interest is the activation pattern that was obtained during the maintenance phase: increasing activation with memory load was found not only in the lateral PFC (BA 9,44) but also in the parietal lobe (BA 7,40), anterior cingulate (BA 32), and cerebellum. Decreasing activation was found in the occipito-temporal lobe (BA 19,39) as well as the medial prefrontal cortex (BA 9,10). Subject increases in pattern expression from 1 to 6 items were positively correlated with the corresponding reaction time increases (p<0.05) and negatively correlated with NARTIQ (p<0.05), indicating that people who were faster in their responses and had higher NARTIQ had to increase their subject expression of the memory-load-related activation pattern less and were more efficient at the cognitive task. Our method thus not only reproduced findings of other WM studies but also addressed the issue of interactions between lateral PFC and other parts of the brain during the task, for the retention of the to-be-remembered information. The load-related activation patterns from encoding and retrieval phase and their relationship to behavior are also discussed. PMID- 15820630 TI - Auditory spatial localization and attention deficits in autistic adults. AB - The objective of this study was to compare autistic adults and matched control subjects in their ability to focus attention selectively on a sound source in a noisy environment. Event-related brain potentials (ERPs) were recorded while subjects attended to a fast paced sequence of brief noise bursts presented in free-field at a central or peripheral location. Competing sequences of noise bursts at adjacent locations were to be ignored. Both behavioral measures of target detection and auditory ERP amplitudes indicated that control subjects were able to focus their attention more sharply on the relevant sound source than autistic subjects. These findings point to a fundamental deficit in the spatial focusing of auditory attention in autism, which may be a factor that impedes social interactions and sensory-guided behavior, particularly in noisy environments. PMID- 15820631 TI - Imagining material versus geometric properties of objects: an fMRI study. AB - Two experiments are reported that used fMRI to compare the brain activation during the imagery of material and geometric object features. In the first experiment, participants were to mentally evaluate objects along either a material dimension (roughness, hardness and temperature; e.g., Which is harder, a potato or a mushroom?) or a geometric dimension (size and shape; e.g., Which is larger, a pumpkin or a cucumber?). In the second experiment, when given the name of an object and either a material (roughness and hardness) or geometric (size and shape) property participants rated the object on a scale from 1 to 4. Both experiments were designed to examine the underlying neural substrate that supports the processing of material object properties with respect to geometric properties. Considering the relative amount of activation across the two types of object properties, we found that (1) the interrogation of geometric features differentially evokes visual imagery which involves the region in and around the intraparietal sulcus, (2) the interrogation of material features differentially evokes the processing of semantic object representations which involves the inferior extrastriate region, and (3) the lateral occipital cortex (LOC) responds to shape processing regardless of whether the feature being queried is a material or geometric feature. PMID- 15820632 TI - An event-related brain potential study of sentence comprehension in preschoolers: semantic and morphosyntactic processing. AB - The goal of this study was to investigate the distinctiveness and the relative time course of the event-related brain potentials (ERP) elicited by syntactically and semantically anomalous words within sentences in 36- and 48-month-old children. ERPs were recorded while children listened to semantically anomalous (i.e., My uncle will blow the movie*), syntactically anomalous (i.e., My uncle will watching the movie*) and control sentences (i.e., My uncle will watch the movie). Semantic violations elicited a negative slow wave with different peaks at 400, 600 and 800 ms in both age groups, whereas the morphosyntactic violations elicited two positive shifts: the first starting at 200 ms with a frontal distribution over the scalp and the second starting at 600 ms and peaking around 800 ms with a broad distribution across the scalp in 36-month-olds and anteriorly distributed for 48-month-olds. These results show that preschoolers display different ERP patterns to syntactic and to semantic violations within sentences. It is possible that the ERP effects here reported are analogous to those elicited in adults by the same type of stimuli, although differences in topography are evident. PMID- 15820633 TI - Differential processing of word and color in unilateral spatial neglect. AB - The aim of the present study was to investigate mechanisms underlying processing of contralesional visual stimuli in brain-damaged patients with unilateral spatial neglect (USN). Nine right-hemisphere-damaged stroke patients with left sided neglect and nine controls performed a reaction-time task involving manual response to a central color patch (target stimulus) flanked to the left or right by a Stroop stimulus they had to ignore. While the word dimension of the flanker affected patients' responses considerably and equally when presented to either side, the color dimension of the flanker had no effect when presented to the left, but had a large effect when presented to the right. Four of the patients performed a control task requiring same/different judgments between either of the two flanker dimensions (color and word) and the central target. Their performance indicated that they were able to process color information from the contralesional field, despite their results in the first experiment. These findings demonstrate a dissociation between how the patients processed different dimensions of the same stimuli and imply that the extent of processing in the contralesional hemifield depends both on task requirements and on the exact features of the stimuli. The implications of these results on normal attentional mechanisms is also discussed. PMID- 15820634 TI - Auditory organization of sound sequences by a temporal or numerical regularity--a mismatch negativity study comparing musicians and non-musicians. AB - The human auditory system can encode regularities in the acoustic environment without the requirement of attention. We investigated whether the auditory system of musicians is more sensitive than that of non-musicians in encoding complex regularities. We presented tone sequences containing either a temporal or a numerical regularity. The sequence with the temporal regularity could be divided into segments of a constant duration while the segments contained a varying number of tones. The sequence with the numerical regularity, on the other hand, could be divided into segments containing a constant number of tones while the segments varied in duration. Auditory encoding of the regularity was determined by measuring whether an occasional segment lengthening, either in time or by number elicited the mismatch negativity (MMN). In both musicians and non musicians, an MMN was elicited when the temporal regularity was violated. In contrast, only in musicians an MMN was elicited to violations of the numerical regularity. The results show that temporal processing is of general importance in audition since at an involuntary auditory processing stage a complex temporal regularity can be encoded irrespective of musical expertise. Furthermore, the auditory system of professional musicians can encode a numerical regularity without attention being required reflecting the functional importance of beat tracking in the perceptual organization of music. PMID- 15820635 TI - Electrical brain responses evoked by human faces in acute psychosis. AB - Patients with schizophrenia are known to have behavioral deficits in recognizing faces and facial expressions. However, the ability to process simple visual stimuli appears to be intact in first-episode psychosis. The aim of this study was to examine complex visual processing, especially the event-related potentials (ERPs) elicited by human faces, in early psychosis. Never-medicated patients in acute psychosis (n=18) were compared with healthy controls (n=19). Photographs of human faces were presented in a classic oddball paradigm requiring a motor response to a smiling face. Cerebral sources of ERPs were analyzed of the averaged responses, using minimum norm estimates, and dipole models. Face sensitive response at 145 ms after the face stimuli was of significantly higher amplitude in our never-medicated patients, and the activity distribution between the groups was clearly different. At the early phase of a psychotic illness, these alterations in face-related neuronal network processes represent perceptual disturbance in psychosis, possibly including state and trait, as well as potential physiological compensatory features. PMID- 15820636 TI - The effects of alpha/theta neurofeedback on personality and mood. AB - Alpha/theta neurofeedback has been shown to be successful both in treating addictions and in enhancing artistry in music students. How its effects are mediated are not yet clear. The present study aimed to test the hypothesis that alpha/theta neurofeedback works inter alia by normalising extreme personality and raising feelings of well being. 12 participants with high scores for Withdrawal (as measured by the PSQ) were given either alpha/theta neurofeedback or mock feedback and their personality and mood were assessed. Withdrawal scores on the PSQ-80 were not found to change in either group but significant effects were found for the Profile Of Mood States (POMS), with real feedback producing higher overall scores than mock feedback (P = 0.056). Real feedback caused participants to feel significantly more energetic (P < 0.01) than did mock feedback. Sessions of real feedback made participants feel more composed (P < 0.01), agreeable (P < 0.01), elevated (P < 0.01) and confident (P < 0.05), whilst sessions of mock feedback made participants feel more tired (P < 0.05), yet composed (P < 0.01). These findings suggest that, whilst 9 sessions of alpha/theta neurofeedback was insufficient to change personality, improvements in mood may provide a partial explanation for the efficacy of alpha/theta neurofeedback. PMID- 15820637 TI - ERP analyses of task effects on semantic processing from words. AB - Semantic (positive) priming refers to the facilitated processing of a probe word when preceded by a related prime word, and is a widely used technique for investigating semantic activation. However, the effect is interrupted or eliminated when attention is directed to low-level features of the prime word, such as its letters, a result which has been used to question the automaticity of semantic processing. We investigated this issue using both behavioural [reaction time (RT)] and electrophysiological measures [event-related potentials (ERPs)]. Subjects performed semantic categorization (living vs. nonliving) and letter search ("A" or "E") tasks on prime words followed by lexical decision on the probe. RT results showed the expected elimination of semantic priming following letter search. However, both prime tasks were affected by the semantic category of the prime, indicating that the meaning was processed. The ERP results supported this conclusion: an early component previously associated with automatic semantic processing [the Recognition Potential (RP)] was sensitive to the category of the prime word irrespective of the prime task. However, a later component (N400) was significantly affected by the task, in both the prime (categorization task) and probe words (semantic priming). The results dissociate rapid, automatic semantic processing from semantic priming. We suggest that a later inhibitory control mechanism suppresses this semantic activation when it is not relevant to the task, and that this produces the loss of semantic priming. PMID- 15820638 TI - P300 recordings during event-related fMRI: a feasibility study. AB - Analysis of combined event-related potentials (ERP) and functional magnetic resonance imaging (fMRI) can provide a high temporal and high spatial resolution to study functional cerebral processes. However, EEG data recorded inside an MR scanner is heavily distorted by artifacts. It is important in cognitive studies to ensure that recorded data reflect the same brain activity, and this is achieved through interleaved electroencephalographic (EEG) and fMRI measurements. Here, we demonstrate the feasibility of recording P300 ERPs during fMRI using a three-stimulus visual oddball task and involving a small number of trials for each stimulus. Ten EEG channels were acquired interleaved with fMRI images in five healthy subjects. The stimuli, including rare targets "X," frequent repetitive distractors "O," and rare distractors referred to as novels, were randomly presented every 2 +/- 1 s. The post hoc filter presented here was designed and applied to EEG data to remove the cardiac pulse artifact. Interleaved EEG/fMRI acquisition evidenced two P300 ERPs evoked at Fz, Cz, and Pz by targets and novels. Novel-related ERPs were of higher amplitude than their target-related counterparts. The fMRI maps acquired concurrently showed stronger BOLD response for target condition. We have shown that interleaved acquisition allows to obtain reliable P300 data and fMRI results, likely to shed light on the anatomical location of brain regions involved in cognitive ERPs relevant to many disorders affecting CNS functions. These noninvasive multimodal neuroimaging techniques can be used to explore and better understand processes underlying the functional brain organization. PMID- 15820639 TI - Spatio-temporal frequency characteristics of intersensory components in audiovisually evoked potentials. AB - Perception of the external world is based on complex neural processes allowing for combination of sensory experiences from different modalities. Audiovisual (AV) integration is discussed in this paper on the basis of the intersensory component (IC), which is the part of the multisensory-evoked potential that is not explained by linear summation of the unisensory-evoked potentials. It was predicted that audiovisual ICs can be extracted, localized, and differentiated by means of wavelet-based frequency analysis. Healthy, right-handed subjects (n = 15) were instructed to view and listen to presented stimuli (A: auditory; V: visual; and AV: audiovisual). Electroencephalographic data was analyzed off-line by means of wavelet transformation utilizing quadratic B-spline mother wavelets. Cross-modal interaction was investigated by subtracting wavelet responses to unimodal stimuli (A, V) alone from the wavelet responses to the combined stimuli (AV; i.e., interaction = AV - (A + V)). These difference waveforms revealed the phase-locked fraction of ICs further characterized by frequency-band and location. Spatio-temporally distinct ICs were observed in all frequency bands [31 62 Hz (gamma), 16-31 Hz (beta), 8-16 Hz (alpha), 4-8 Hz (theta), 0.5-4 Hz band (delta)]. These were most pronounced and sustained in the theta frequency band with early (<100 ms) appearance in fronto-centro-parietal sites. In contrast, alpha-band ICs tended to appear later (>200 ms) in these locations. High frequency (beta- and gamma-band) ICs were less organized in their spatial pattern with both early and late appearance. ICs may reflect sensory and cognitive/integrative processes at the cortical level. In case of intersensory processing, alpha- and theta-activity appear to be spatio-temporally distinct, and could therefore participate in different stages of perception. These findings add further support to current model views of oscillatory activity in selectively distributed networks. PMID- 15820640 TI - Facial expression decoding in early Parkinson's disease. AB - The ability to derive emotional and non-emotional information from unfamiliar, static faces was evaluated in 21 adults with idiopathic Parkinson's disease (PD) and 21 healthy control subjects. Participants' sensitivity to emotional expressions was comprehensively assessed in tasks of discrimination, identification, and rating of five basic emotions: happiness, (pleasant) surprise, anger, disgust, and sadness. Subjects also discriminated and identified faces according to underlying phonemic ("facial speech") cues and completed a neuropsychological test battery. Results uncovered limited evidence that the processing of emotional faces differed between the two groups in our various conditions, adding to recent arguments that these skills are frequently intact in non-demented adults with PD [R. Adolphs, R. Schul, D. Tranel, Intact recognition of facial emotion in Parkinson's disease, Neuropsychology 12 (1998) 253-258]. Patients could also accurately interpret facial speech cues and discriminate the identity of unfamiliar faces in a normal manner. There were some indications that basal ganglia pathology in PD contributed to selective difficulties recognizing facial expressions of disgust, consistent with a growing literature on this topic. Collectively, findings argue that abnormalities for face processing are not a consistent or generalized feature of medicated adults with mild-moderate PD, prompting discussion of issues that may be contributing to heterogeneity within this literature. Our results imply a more limited role for the basal ganglia in the processing of emotion from static faces relative to speech prosody, for which the same PD patients exhibited pronounced deficits in a parallel set of tasks [M.D. Pell, C. Leonard, Processing emotional tone from speech in Parkinson's disease: a role for the basal ganglia, Cogn. Affect. Behav. Neurosci. 3 (2003) 275-288]. These diverging patterns allow for the possibility that basal ganglia mechanisms are more engaged by temporally-encoded social information derived from cue sequences over time. PMID- 15820641 TI - Event-related potentials reveal dissociable mechanisms for orienting and focusing visuospatial attention. AB - The neural mechanisms supporting visuospatial orienting and focusing were investigated by recording event-related potentials (ERPs) in a cued, line orientation discrimination task. Search arrays flashed randomly in the left or right visual field and were preceded by peripheral cues that varied in validity (valid or invalid, with 50% each) and size (large or small, with 50% each). Facilitation of response time was observed for valid trials, regardless of cue size. In contrast to previous cued search studies, however, small (i.e., more precise) cues were associated with delayed responses. Both the timing and the amplitudes of the early ERP components, P1 and N1, were modulated by attentional orienting, with valid trials eliciting a larger and later contralateral vP1 (ventral P1) and a smaller and later contralateral N1 compared to invalid trials. Attentional focusing modulated only the amplitudes of the P1 component, with precisely cued trials eliciting a larger dP1 (dorsal P1) than less precisely cued trials at both contralateral and ipsilateral sites. Thus, both attentional orienting and focusing modulate early stimulus processing stages that overlap in time, but with dissociable effects on the scalp distribution of these components, indicating possibly different underlying mechanisms. In addition, the results support the notion that voluntary and involuntary allocations of visuospatial attention are mediated by different underlying neural processes. PMID- 15820642 TI - Gating of SEPs by contraction of the contralateral homologous muscle during the preparatory period of self-initiated plantar flexion. AB - To investigate the centrifugal change in somatosensory information processing caused by contraction of the contralateral homologous muscle, we recorded the somatosensory-evoked potentials (SEPs) during the preparatory period of a self initiated plantar flexion. The SEPs following stimulation of the right tibial nerve at the popliteal fossa were recorded in nine healthy subjects. Self initiated plantar flexion of the left ankle was performed once every 5 to 7 s. The electrical stimulation was delivered continuously, and the subjects were instructed to concentrate on the movement and not to pay attention to the electrical stimulation. Based on the components of movement-related cortical potential, Bereitschaftspotential (BP) and Negative slope (NS), the preparatory period was divided into four sub-periods (NS, BP-1, BP-2, and Pre-BP). To obtain pre-movement SEPs, the signals following stimulation in each sub-period were averaged. SEPs were attenuated in the preparatory period, especially in the NS sub-period. The amplitude of N40 component was significantly attenuated compared with that in the stationary state and other sub-periods. The amplitude of P53 and N70 was smaller in the NS sub-period than other pre-movement sub-periods. Since there was no centripetal effect on SEPs in the preparatory period, these findings suggested that the activity of motor-related areas modulated the somatosensory information from the contralateral non-movement limb (centrifugal gating). It was assumed that an inhibition on the somatosensory inputs from contralateral limb was caused by the projection via either the corpus callosum or ipsilateral cortico-cortical projections. PMID- 15820644 TI - Programming effectors and coordination in bimanual in-phase mirror finger movements. AB - We investigated cerebral activation during programming of in-phase symmetric finger movements in a precued response task. Partial precues provided advance information about either mirror effectors or in-phase coordination of bimanual movements, while full precue specified both response parameters and neutral precue no movement information. Effects of precueing were assessed on reaction time (RT), contingent negative variation (CNV), and alpha and beta event-related desynchronization (ERD). Information on coordination mode induced less efficient preparation than information on effectors, as revealed by longer RT, but paradoxically the CNV was found of larger amplitude for in-phase than for mirror precue. Full and in-phase precues were associated to largest cerebral activation, as reflected by CNV amplitude as well as beta ERD. It is suggested that with in phase precueing, abstract programming of coordination and concrete preparation of possible effectors overlap, engaging more cerebral resources than when symmetric effectors are pre-specified. Alpha ERD underwent regional modulations dependent on the type of preparation, pointing out the role of the right parietal region in visuomotor transformation with full movement programming, and the preferential implication of the dominant hemisphere and medial brain regions in synchronization of both hand movements. Beta ERD topographical distribution suggested an increased implication of bilateral and medial motor regions in anticipation to the response signal with incomplete movement preparation. PMID- 15820643 TI - Distinctions between manipulation and function knowledge of objects: evidence from functional magnetic resonance imaging. AB - A prominent account of conceptual knowledge proposes that information is distributed over visual, tactile, auditory, motor and verbal-declarative attribute domains to the degree to which these features were activated when the knowledge was acquired [D.A. Allport, Distributed memory, modular subsystems and dysphagia, In: S.K. Newman, R. Epstein (Eds.), Current perspectives in dysphagia, Churchill Livingstone, Edinburgh, 1985, pp. 32-60]. A corollary is that when drawing upon this knowledge (e.g., to answer questions), particular aspects of this distributed information is re-activated as a function of the requirements of the task at hand [L.J. Buxbaum, E.M. Saffran, Knowledge of object manipulation and object function: dissociations in apraxic and non-apraxic subjects. Brain and Language, 82 (2002) 179-199; L.J. Buxbaum, T. Veramonti, M.F. Schwartz, Function and manipulation tool knowledge in apraxia: knowing 'what for' but not 'how', Neurocase, 6 (2000) 83-97; W. Simmons, L. Barsalou, The similarity-in-topography principle: Reconciling theories of conceptual deficits, Cognitive Neuropsychology, 20 (2003) 451-486]. This account predicts that answering questions about object manipulation should activate brain regions previously identified as components of the distributed sensory-motor system involved in object use, whereas answering questions about object function (that is, the purpose that it serves) should activate regions identified as components of the systems supporting verbal-declarative features. These predictions were tested in a functional magnetic resonance imaging (fMRI) study in which 15 participants viewed picture or word pairs denoting manipulable objects and determined whether the objects are manipulated similarly (M condition) or serve the same function (F condition). Significantly greater and more extensive activations in the left inferior parietal lobe bordering the intraparietal sulcus were seen in the M condition with pictures and, to a lesser degree, words. These findings are consistent with the known role of this region in skilled object use [K.M. Heilman, L.J. Gonzalez Rothi, Apraxia, In: K.M. Heilman, E. Valenstein (Eds.), Clinical Neuropsychology, Oxford University Press, New York, 1993, pp. 141-150] as well as previous fMRI results [M. Kellenbach, M. Brett, K. Patterson, Actions speak louder than functions: the importance of manipulability and action in tool representation, Journal of Cognitive Neuroscience, 15 (2003) 30-46] and behavioral findings in brain-lesion patients [L.J. Buxbaum, E.M. Saffran, Knowledge of object manipulation and object function: dissociations in apraxic and non-apraxic subjects, Brain and Language, 82 (2002) 179-199]. No brain regions were significantly more activated in the F than M condition. These data suggest that brain regions specialized for sensory-motor function are a critical component of distributed representations of manipulable objects. PMID- 15820645 TI - Active processing of biological motion perception: an ERP study. AB - The purpose of this study was to measure event-related potentials (ERPs) to clarify how attention affects neural activity during the visual perception of biological motion (BM). Thirteen healthy subjects observed BM or scrambled motion (SM). For SM, each point had the same velocity vector as in BM, but the initial starting positions were randomized. Each BM and SM was overlaid with ten noise dots and four rectangles. For the rectangles, one was or was not rotated 90 degrees relative to the others. Subjects were required to undertake two kinds of visual tasks. For the attention-to-motion condition, subjects directed their attention to the type of motion. For the attention-to-rectangle condition, subjects directed their attention to the rotational angle of the overlaid rectangles. As in our earlier study, the ERP response to the perception of BM had two negative components at approximately 200 ms (N200) and approximately 330 ms (N330) in both attentional conditions. Our analysis focused on the amplitude of the second negative component, which was sensitive to BM. There was a significant interaction between attention and the type of motion regarding the amplitude of N330. Specifically, the amplitude of N330 in response to BM was greater in the attention-to-motion condition than in the attention-to-rectangle condition, and was greater than the amplitude of the response to SM in the attention-to-motion condition. These results suggest that in this experimental design, processing of BM is modulated by attention. PMID- 15820646 TI - Cortical topography of human anterior intraparietal cortex active during visually guided grasping. AB - Dexterous manual prehension requires successfully transforming sensory representations of an object's intrinsic spatial properties (e.g., shape) into motor plans for configuring the opposition space of the hand. In macaques, these sensorimotor transformations are accomplished in a circuit connecting the anterior intraparietal sulcus (area AIP) with inferior frontal cortex (area F5ab). Activation in the human anterior intraparietal sulcus (aIPS) during visually guided grasping suggests a homologue of macaque area AIP. If true, then despite individual variation in cortical topography, visually guided grasping should be consistently associated with focal activation at the junction of the IPS and postcentral sulcus. FMRI was used to test this hypothesis in 14 right handed adults. Despite substantial variability in IPS topography, a contrast between pincer grasping vs. reaching to complex asymmetrical shapes revealed activation foci at the junction of the IPS and postcentral gyrus in all 14 individuals. This site is likely within the most superior, rostral aspect of Brodmann's area 40, corresponding to area PF or PDE as defined by von Economo and Koskinas, and area 86 as defined by Vogt and colleagues. In both humans and macaques this region appears to play a key role in visually guided grasping. PMID- 15820647 TI - Influence of gaze direction on crossmodal modulation of visual ERPS by endogenous tactile spatial attention. AB - Recent evidence indicates that the spatial direction of endogenous covert spatial attention in one sensory modality can crossmodally influence early processing of stimuli in a different modality. However, spatial locations are initially coded according to different frames of reference for different modalities (e.g., body centered for touch versus retinocentric vision) and postural changes (e.g., gaze shifts) will realign these. Here, we used event-related potentials (ERPs) to investigate how the direction of endogenous tactile attention affects sensory specific visual ERP components. Critically, by manipulating direction of gaze, we were able to test whether any crossmodal effects depend on visual and tactile projections to a common hemisphere, on common locations in external space, or on some combination of the two. We found that both P1 and N1 visual components were modulated according to the direction of endogenous tactile attention. While the P1 crossmodal effect followed purely hemispheric constraints, the attentional modulation of N1 appeared to combine both anatomical and external spatial constraints. PMID- 15820648 TI - What is in a name: comparing the Tower of London with one of its variants. AB - Since the implementation of the Tower of London (ToL) test by Shallice in 1982, numerous variants differing in the tower's physical appearance have been developed. Here we compare behavioral performance (n = 31) on the original Tower of London task consisting of three rods of unequal lengths with a three-ball version of the Ward and Allport Tower Task (1997) using three equally sized rods. In the problem set used, the start and goal states for both tower configurations were identical across all trials. The experiment was divided into two parts: in the first block, the problems presented were equalized with respect to the number of paths for achieving an optimal solution, the minimum number of moves, goal hierarchy, subgoaling patterns, and suboptimal alternatives between the two tower versions. As expected, participants showed the same performance scores for both types of towers when structural problem parameters were identical. In the second block, participants had to solve five-move problems which-due to the different rod sizes of the towers-had only one optimal solution in the original version, but two optimal solutions in the variant with three rods of equal length. Participants revealed lower performance scores and showed longer planning times in the original version than in the second tower version. These findings demonstrate that the two tower versions are only interchangeable when specific planning parameters are equalized. Otherwise, even if problems look identical, significant differences in performance may be found due to the differing problem spaces in the two tower versions. PMID- 15820649 TI - Seeing and hearing others and oneself talk. AB - We studied the modification of auditory perception in three different conditions in twenty subjects. Observing other person's discordant articulatory gestures deteriorated identification of acoustic speech stimuli and modified the auditory percept, causing a strong McGurk effect. A similar effect was found when the subjects watched their own silent articulation in a mirror and acoustic stimuli were simultaneously presented to their ears. Interestingly, a smaller but significant effect was even obtained when the subjects just silently articulated the syllables without visual feedback. On the other hand, observing other person's or one's own concordant articulation and silently articulating a concordant syllable improved identification of the acoustic stimuli. The modification of auditory percepts caused by visual observation of speech and silently articulating it are both suggested to be due to the alteration of activity in the auditory cortex. Our findings support the idea of a close relationship between speech perception and production. PMID- 15820650 TI - Coordination of bowing and fingering in violin playing. AB - Playing string instruments implies motor skills including asymmetrical interlimb coordination. How special is musical skill as compared to other bimanually coordinated, non-musical skillful performances? We succeeded for the first time to measure quantitatively bimanual coordination in violinists playing repeatedly a simple tone sequence. A motion analysis system was used to record finger and bow trajectories for assessing the temporal structure of finger-press, finger lift (left hand), and bow stroke reversals (right arm). The main results were: (1) fingering consisted of serial and parallel (anticipatory) mechanisms; (2) synchronization between finger and bow actions varied from -12 ms to 60 ms, but these 'errors' were not perceived. The results suggest that (1) bow-finger synchronization varied by about 50 ms from perfect simultaneity, but without impairing auditory perception; (2) the temporal structure depends on a number of combinatorial mechanisms of bowing and fingering. These basic mechanisms were observed in all players, including all amateurs. The successful biomechanical measures of fingering and bowing open a vast practical field of assessing motor skills. Thus, objective assessment of larger groups of string players with varying musical proficiency, or of professional string players developing movement disorders, may be helpful in music education. PMID- 15820651 TI - The extent of visual deficit and auditory spatial compensation: evidence from self-positioning from auditory cues. AB - Blindfolded sighted, myopic, amblyopic, adventitiously blind and congenitally blind humans performed a self-positioning task during which they were stimulated only by auditory cues. Results showed that visually deprived subjects used auditory cues to position themselves in their environment with a greater accuracy than normal-sighted subjects. In addition, the magnitude of auditory spatial compensation was found to be strongly related to the extent of the visual deficit. PMID- 15820652 TI - Contagious yawning and the brain. AB - Contagious yawning, the onset of a yawn triggered by seeing, hearing, reading, or thinking about another person yawn is a well-documented phenomenon. The mechanisms that drive contagious yawning are as yet unknown, but there is recent evidence of a link between contagious yawning and self-processing (S.M. Platek, S.R. Critton, T.E. Myers, G.G. Gallup Jr., Contagious yawning: the role of self awareness and mental state attribution, Cogn. Brain Res. 17 (2003) 223-227.) that is negatively impacted by schizotypal personality traits. The neural substrates involved in contagious yawning, however, are unknown. Here, using fMRI, we show that viewing someone yawn evokes unique neural activity in the posterior cingulate and precuneus. Because of the role these areas play in self-processing (e.g., self-referential processing, theory of mind, autobiographical memory), our findings provide further support for the hypothesis that contagious yawning may be part of a neural network involved in empathy. PMID- 15820653 TI - Are perceptual judgments dissociated from motor processes?--A prism adaptation study. AB - When asked to choose which of two chimeric faces composed of 'smiling' and 'neutral' half-faces is happier, healthy adults select the face in which the left half is smiling. Here, we show that this perceptual leftward bias is associated with a bias in eye movements to the same side. However, when we shifted the pattern of eye movements towards the right side by using prismatic lenses, we did not observe a concurrent shift in the perceptual judgments. Therefore, we argue that overt motor responses are not necessary for perceptual judgments. Furthermore, we argue that while prism adaptation influences performance on motor tasks, it cannot influence higher-order representational processes. PMID- 15820654 TI - I've heard it all before: perceptual invariance represented by early cortical auditory-evoked responses. AB - Sensitivity to acoustic invariance is critical for establishing stable representations in a shifting world of sound. By recording early auditory cortical responses to complex sounds in human listeners and categorising these responses according to the maintenance or change of stimulus attributes across consecutive presentations, we show that repetition within a constantly varying acoustic context produces enhanced neural responding in auditory cortices. PMID- 15820655 TI - Effects of phosphate limitation on expression of genes involved in pyrimidine synthesis and salvaging in Arabidopsis. AB - Arabidopsis seedlings grown for 14 d without phosphate (P) exhibited stunted growth and other visible symptoms associated with P deficiency. RNA contents in shoots decreased nearly 90%, relative to controls. In shoots, expression of Pht1;2, encoding an inducible high-affinity phosphate transporter, increased threefold, compared with controls, and served as a molecular marker for P limitation. Transcript levels for five enzymes (aspartate transcarbamoylase, ATCase, EC 2.1.3.2; carbamoyl phosphate synthetase, CPSase, EC 6.3.5.5); UMP synthase, EC 2.4.1.10, EC 4.1.1.23; uracil phosphoribosyltransferase, UPRTase, EC 2.4.2.9; UMP kinase, EC 2.7.1.14) increased 2-10-fold in response to P starvation in shoots. These enzymes, which utilize phosphorylated intermediates at putative regulated steps in de novo synthesis and salvaging pathways leading to UMP and pyrimidine nucleotide formation, appear to be coordinately regulated, at the level of gene expression. This response may facilitate pyrimidine nucleotide synthesis under P limitation in this plant. Expression of P-dependent and P independent phosphoribosyl pyrophosphate (PRPP) synthases (PRS2 and PRS3, respectively) which provide PRPP, the phosphoribosyl donor in UMP synthesis via both de novo and salvaging pathways, was differentially regulated in response to P limitation. PRS2 mRNA levels increased twofold in roots and shoots of P-starved plants, while PRS3 was constitutively-expressed. PRS3 may play a novel role in providing PRPP to cellular metabolism under low P availability. PMID- 15820656 TI - Ethylene induced shikonin biosynthesis in shoot culture of Lithospermum erythrorhizon. AB - Lithospermum erythrorhizon shoots, cultured on phytohormone-free Murashige and Skoog solid medium, produced shikonin derivatives, whereas shoots cultured in well-ventilated petri dishes, produced small amount. Analysis by gas chromatography revealed the presence of ethylene in non-ventilated petri dishes where the shoots, producing shikonin derivatives, were cultured. Therefore, the possible involvement of ethylene in shikonin biosynthesis of shoot cultures was investigated. Treatment of ethylene or the ethylene precursor, 1 aminocyclopropane-1-carboxylic acid, resulted in increasing shikonin derivatives contents in cultured shoots. Silver ion, an ethylene-response inhibitor, or aminoethoxyvinylglycine, an ethylene biosynthesis inhibitor, decreased production of shikonin derivatives in cultured shoots. Our results indicate that ethylene is one of the regulatory elements of shikonin biosynthesis in L. erythrorhizon shoot culture. PMID- 15820657 TI - Modulation of cadmium-induced oxidative stress in Ceratophyllum demersum by zinc involves ascorbate-glutathione cycle and glutathione metabolism. AB - To understand the interaction between Zn, an essential micronutrient and Cd, a non-essential element, Cd-10 microM and Zn supplemented (10, 50, 100, and 200 microM) Cd 10 microM treated Ceratophyllum demersum L. (Coontail), a free floating freshwater macrophyte was chosen for the study. Cadmium at 10 microM concentration decreased thiol content, enhanced oxidation of ascorbate (AsA) and glutathione (GSH) to dehydroascorbate (DHA) and glutathione disulfide (GSSG), respectively, a clear indication of oxidative stress. Zinc supplementation to Cd (10 microM) treated plants effectively restored thiols, inhibited oxidation of AsA and GSH maintaining the redox molecules in reduced form. Cd-10 microM slightly induced ascorbate peroxidase (APX, E.C. 1.11.1.11) but inhibited monodehydroascorbate reductase (MDHAR, E.C. 1.6.5.4), dehydroascorbate reductase (DHAR, E.C. 1.8.5.1) and glutathione reductase (GR, E.C. 1.6.4.2), enzymes of ascorbate-glutathione cycle (AGC). Zn supplementation restored and enhanced the functional activity of all the AGC enzymes (APX, MDHAR, DHAR and GR). Gamma glutamylcysteine synthetase (gamma-GCS, E.C. 6.3.2.2) was not affected by Cd as well as Zn, but Zn supplements increased glutathione-S-transferase (GST, E.C. 2.5.1.18) activity to a greater extent than Cd and simultaneously restored glutathione peroxidase (GSH-PX, E.C. 1.11.1.9) activity impaired by Cd toxicity. Zn-alone treatments did not change above investigated parameters. These results clearly indicate the protective role of Zn in modulating the redox status of the plant system through the antioxidant pathway AGC and GSH metabolic enzymes for combating Cd induced oxidative stress. PMID- 15820658 TI - Free radical generation in Pinus sylvestris and Larix decidua seeds primed with polyethylene glycol or potassium salt solutions. AB - Electron paramagnetic resonance (EPR) spectra of Pinus sylvestris and Larix decidua seeds show that priming with PEG+200 mg kg(-1) gibberelic acid (GA(3)) results in appreciably higher free radical contents than in unprimed control samples. Only relatively minor changes in the free radical levels were observed in seeds primed with K(+) salts. However, both priming treatments have been reported previously to result in faster germination rates compared to controls without changing the germination percentage. In measurements on individual seeds of L. decidua, there were no significant differences between the mean free radical levels in viable and non-viable seeds within each treatment group. Thus, the elevation in free radical levels in the PEG+GA(3) treatments appear to be a direct consequence of the priming treatment and do not correspond to the initiation of germination. PMID- 15820659 TI - Methyl jasmonate modulated biotransformation of phenylpropanoids to vanillin related metabolites using Capsicum frutescens root cultures. AB - Normal root cultures of Capsicum frutescens biotransform externally fed precursors, like caffeic acid and veratraldehyde, to vanillin and other related metabolites. The bioconversion of caffeic acid to further metabolites--viz. vanillin, vanillylamine, vanillic acid--was shown to be elicited by treating the cultures with 10 microM methyl jasmonate (MJ). Root cultures treated with MJ accumulated (1.93 times) more of vanillin (20.2 microM on day-3) than untreated ones. A concomitant increase in enzymatic activity of caffeic acid O-methyl transferase (CAOMT, EC 2.1.1.68) was obtained in MJ treated cultures, compared to untreated cultures. After 24 h of MJ treatment, a 13.7-fold increase in CAOMT activity was recorded in root cultures of C. frutescens. Cultures treated with veratraldehyde accumulated more vanillin (78 microM) than caffeic acid fed cultures, 6 days after precursor addition. Capsaicin did not accumulate even after addition of precursors. The efficiencies of biotransformation with caffeic acid and veratraldehyde were 2.2% and 9% with respect to vanillin formation, indicating a possible diversion of the phenylpropanoid pathway towards other secondary metabolites. PMID- 15820660 TI - Sucrose may play an additional role to that of an osmolyte in Synechocystis sp. PCC 6803 salt-shocked cells. AB - The role of sucrose in cyanobacteria is still not fully understood. It is generally considered a salt-response molecule, and particularly, in Synechocystis sp. strain PCC 6803, it is referred as a secondary osmolyte. We showed that sucrose accumulates transiently in Synechocystis cells at early stages of a salt shock, which could be ascribed to salt activation of sucrose-phosphate synthase (SPS, UDP-glucose: D-fructose-6-phosphate 2-alpha-D-glucosyltransferase; EC 2.4.1.14), the key enzyme in sucrose synthesis pathway, and to an increase of the expression of the SPS encoding gene. Experiments with a mutant strain impaired in sucrose biosynthesis showed that sucrose is essential in stationary phase cells to overcome a later salt stress. Taken together, these results led us to suggest a more intricate function for sucrose than to be an osmoprotectant compound. PMID- 15820661 TI - Enhanced tolerance to salt stress in transgenic loblolly pine simultaneously expressing two genes encoding mannitol-1-phosphate dehydrogenase and glucitol-6 phosphate dehydrogenase. AB - A reproducible approach to improve salt tolerance of conifers has been established by using the technology of plant genetic transformation and using loblolly pine (Pinus taeda L.) as a model plant. Mature zygotic embryos of three genotypes of loblolly pine were infected with Agrobacterium tumefaciens strain LBA 4404 harboring the plasmid pBIGM which carrying two bacterial genes encoding the mannitol-1-phosphate dehydrogenase (Mt1D, EC 1.1.1.17) and glucitol-6 phosphate dehydrogenase (GutD) (EC 1.1.1.140), respectively. Transgenic plantlets were produced on selection medium containing 15 mg l(-1) kanamycin and confirmed by polymerase chain reaction (PCR) and Southern blot analysis of genomic DNA. The Mt1D and GutD genes were expressed and translated into functional enzymes that resulted in the synthesis and accumulation of mannitol and glucitol in transgenic plants. Salt tolerance assays demonstrated that transgenic plantlets producing mannitol and glucitol had an increased ability to tolerate high salinity. These results suggested that an efficient A. tumefaciens-mediated transformation protocol for stable integration of bacterial Mt1D and GutD genes into loblolly pine has been developed and this could be useful for the future studies on engineering breeding of conifers. PMID- 15820662 TI - Effects of elevated pCO2 and/or pO3 on C-, N-, and S-metabolites in the leaves of juvenile beech and spruce differ between trees grown in monoculture and mixed culture. AB - Three and four-year-old saplings of beech (Fagus sylvatica L.) and spruce (Picea abies (L.) Karst.) grown in monoculture and mixed culture were exposed in phytotrons to (1) ambient air, (2) elevated pO(3), (3) elevated pCO(2), or (4) elevated pCO(2) plus elevated pO(3). After 5 months, the contents of soluble sugars, starch, soluble amino compounds, non-structural proteins (NSP), as well as reduced (GSH) and oxidized (GSSG) glutathione were determined in the leaves of both species in order to assess the effects of the gaseous regimes on primary metabolism. Elevated pO(3) did not affect sugar and starch levels in beech leaves in monoculture, but significantly increased sugar levels in beech leaves grown in mixed culture. In spruce needles, sugar levels tended to be enhanced in both culture types. Individual and combined exposure of elevated pCO(2) led to an increase in non-structural carbohydrate (soluble sugars plus starch) levels in beech and spruce leaves of both culture types. Differences in the responses of non-structural carbohydrate levels to elevated pCO(2) between beech and spruce were apparent from different contributions of sugars and starch to the increase in carbohydrate levels. Exposure to elevated pCO(2) and/or elevated pO(3) did not affect the levels of soluble amino compounds and NSP in beech leaves, but reduced amino compound levels in spruce needles of both culture types. Elevated pO(3) increased GSH levels in the leaves of both tree species in both culture types, while GSSG levels in monoculture were reduced in beech leaves, but significantly enhanced in spruce needles. Elevated pCO(2) reduced GSSG levels in beech and spruce leaves in monoculture, and GSH levels in spruce needles of both culture types. The combination of elevated pCO(2) and pO(3) increased GSSG levels in beech leaves of both culture types and in spruce needles in monoculture, but reduced GSH levels in spruce needles of both culture types. Apparently, under each gaseous regime, the culture type significantly altered primary metabolism of the leaves of beech and spruce. PMID- 15820663 TI - Pectin methyl esterases and pectins in normal and hyperhydric shoots of carnation cultured in vitro. AB - Control and hyperhydric micropropagated plantlets from three carnation cultivars have been used to study their pectin composition and the activity of pectin methyl esterases (PMEs; EC 3.1.1.11). Pectins are a highly heterogeneous group of polymers that contribute to cell adhesion, cell wall architecture, and cell wall mechanical strength. Pectins control cell wall porosity and cell wall ionic status and are implicated in intercellular space development. The degree of esterification of pectins is controlled by the activity of cell wall PMEs; their different actions can affect the properties of the cell wall, which have been considered important with respect to controlling the development of hyperhydricity. The total pectins of hyperhydric leaves of the three varieties were significantly reduced in comparison with controls. The pectate fraction was significantly increased in hyperhydric leaves of all varieties while soluble pectins and protopectins were significantly lower. The PME activity of hyperhydric leaves was higher (4-10 times) compared to controls of the three varieties. Isoelectric focusing of PME isozymes revealed the presence of three isoforms; neutral PME activity was the major isozyme in control and hyperhydric leaves of the three varieties, whilst a decrease in the activity of the acidic isoforms was observed in hyperhydric leaves. The different PME activities could regulate some of the structural changes related to hyperhydricity in micropropagated carnation plants. PMID- 15820664 TI - Effect of the application of benzyladenine pulse on organogenesis, acclimatisation and endogenous phytohormone content in kiwi explants cultured under autotrophic conditions. AB - In traditional in vitro culture, explants grow enclosed in a non-ventilated vessel at high relative humidity with phytohormones continuously present and sucrose as the main energy source. Under such conditions explant growth is far from normal. In this paper, explants of Actinidia deliciosa were cultured in MS medium supplemented with sucrose, benzyladenine and gibberellic acid under autotrophic conditions in glass boxes flushed with air enriched with 600 microl l(-1) CO(2) for the first 20 days and then transferred to MS medium until the end of the culture period. The effect of benzyladenine was assayed in two regimes of application: in cultures for 20 days in the medium or only 24 h in the presence of benzyladenine with the aim of improving shoot proliferation and acclimatisation. The longest explants were those grown under ventilation and pulsed for 24 h with benzyladenine. These explants also rooted spontaneously, whereas those grown with continuous benzyladenine under ventilation or without ventilation grew and rooted poorly. The highest amount of endogenous isoprenoid cytokinins were found in the longest explants grown under ventilation and pulsed for 24 h with benzyladenine; under these conditions zeatin riboside represented two thirds of the entire cytokinin pool. These explants presented the highest amount of indole-3-acetic acid, while abscisic acid content was high in explants cultured under non-ventilated conditions. No differences were observed between explants cultured under ventilation regardless of their exposure to benzyladenine. The longest explants, which also performed best in acclimatisation, also presented a high indole-3-acetic to abscisic acid ratio. PMID- 15820665 TI - The gene for a xyloglucan endotransglucosylase/hydrolase from Cicer arietinum is strongly expressed in elongating tissues. AB - We have isolated a Cicer arietinum cDNA clone (CaXTH1) encoding a protein that belongs to the family 16 of glycosyl hydrolases and has all the conserved features of xyloglucan endotransglucosylase/hydrolases (XTH) proteins, including the presence of a highly conserved domain (DEIDFEFLG) and four Cys which suggest the potential for forming disulfide bonds. These facts indicate that CaXTH1 encodes a putative XTH. This chickpea protein showed a high level of sequence identity with group 1 XTHs that have xyloglucan endotransglucosylase (XET) activity. CaXTH1 was selected by differential screening of a cDNA library constructed using mRNA from C. arietinum polyethylene glycol (PEG) treated epicotyls, as a clone whose expression decreased when epicotyl growth was inhibited by PEG. CaXTH1 shows an expression pattern that seems to be specific for growing tissue, mostly epicotyls and the growing internodes of adult stems. CaXTH1 mRNA was not detected in any other organs of either seedlings or adult plants. CaXTH1 mRNA was abundant when epicotyls are actively growing; there was almost no expression after PEG-treatment. CaXTH1 was up-regulated by indole acetic acid (IAA) and brassinolides (BR), showing the highest transcript levels after IAA plus BR treatment. In situ hybridization study revealed that CaXTH1 is mainly expressed in epidermal cells, the target of the cell expansion process, and also in vascular tissues. The present results suggest an involvement of the putative XTH encoded by CaXTH1 in the chickpea cell expansion process. PMID- 15820666 TI - Expression of multiple forms of polygalacturonase gene during ripening in banana fruit. AB - The activity of polygalacturonase (PG, E.C 3.2.1.15) during ripening in climacteric fruits has been positively correlated with softening of the fruit tissue and differential expression of its gene is suspected to be regulated by the plant hormone ethylene. We have cloned four partial cDNAs, MAPG1 (acc. no. AF311881), MAPG2 (acc. no. AF311882), MAPG3 (acc. no. AF542382) and MAPG4 (acc. no. AY603341) for PG genes and studied their differential expression during ripening in banana. MAPG3 and MAPG4 are believed to be ripening related and regulated by ethylene whereas MAPG2 is associated more with senescence. MAPG1 shows constitutive expression and is not significantly expressed in fruit tissue. The genomic clone MAGPG (acc. No. AY603340) includes the complete MAPG3 gene, which consists of four exons and three introns. The structure of the gene has more similarity to tomato abscission PG rather than tomato fruit PG. It is concluded that softening during ripening in banana fruit results from the concerted action of at least four PG genes, which are differentially expressed during ripening. PMID- 15820667 TI - Characterization of a jasmonate-regulated wheat protein related to a beta glucosidase-aggregating factor. AB - Jasmonates are distributed throughout higher plants, where they play an important role in the activation of signal transduction pathways in response to wounding and pathogen attack. Jasmonates are known to induce the transcriptional activation of plant defense genes, produce jasmonate-regulated proteins (JRP). One class of 32 kDa JRP (designated as JRP-32 in this paper) is present in the Gramineae family, although the function of these proteins is still unclear. A cDNA was cloned from wheat (Triticum aestivum L.) and designated as Ta-JA1. Sequence comparison indicated that Ta-JA1 encodes a JRP-32 protein. Ta-JA1 exists as a single-copy gene, but other genes with low sequence similarity to Ta-JA1 may be present in the wheat genome. The expression of Ta-JA1 was confined to stem tissues and barely detected in leaf and root tissues. Western blot analyses demonstrated that the recombinant Ta-JA1 protein cross-reacts with maize beta glucosidase-aggregating factor (BGAF) antibody. Molecular modeling showed that Ta JA1 and BGAF have a very similar three-dimensional structure. Protein structure analysis indicated that Ta-JA1 together with some related proteins (maize BGAF, wheat Ver2, WCI-1 and Hfr-1) contain two functional domains: a disease response domain and jacalin-related lectin (JRL) domain. A mannose-binding site was also well conserved in these proteins. The data support the hypothesis that JRP-32 and related proteins from Gramineae form a small protein family related to JRLs. This small protein family may have evolved from mannose-specific jacalin-related lectins (mJRLs) by developing a disease response domain in their N-terminus, which may have broadened the functional role of these proteins to include the plant defense response. PMID- 15820668 TI - Differential changes in degradation of chlorophyll-protein complexes of photosystem I and photosystem II during flag leaf senescence of rice. AB - The stability of chlorophyll-protein complexes of photosystem I (PSI) and photosystem II (PSII) was investigated by chlorophyll (Chl) fluorescence spectroscopy, absorption spectra and native green gel separation system during flag leaf senescence of two rice varieties (IIyou 129 and Shanyou 63) grown under outdoor conditions. During leaf senescence, photosynthetic CO(2) assimilation rate, carboxylase activity of Rubisco, chlorophyll and carotenoids contents, and the chlorophyll a/b ratio decreased significantly. The 77 K Chl fluorescence emission spectra of thylakoid membranes from mature leaves had two peaks at around 685 and 735 nm emitting mainly from PSII and PSI, respectively. The total Chl fluorescence yields of PSI and PSII decreased significantly with senescence progressing. However, the decrease in the Chl fluorescence yield of PSI was greater than in the yield of PSII, suggesting that the rate of degradation in chlorophyll-protein complexes of PSI was greater than in chlorophyll-protein complexes of PSII. The fluorescence yields for all chlorophyll-protein complexes decreased significantly with leaf senescence in two rice varieties but the extents of their decrease were significantly different. The greatest decrease in the Chl fluorescence yield was in PSI core, followed by LHCI, CP47, CP43, and LHCII. These results indicate that the rate of degradation for each chlorophyll protein complex was different and the order for the stability of chlorophyll protein complexes during leaf senescence was: LHCII>CP43>CP47>LHCI>PSI core, which was partly supported by the green gel electrophoresis of the chlorophyll protein complexes. PMID- 15820669 TI - Selective memory impairment for public events in a patient with left temporal lobe epilepsy. AB - Highly selective memory impairment for public events was demonstrated in a patient (JR), who suffered from temporal lobe epilepsy (TLE). We successfully trained JR's memory for a set of news events and discuss, on those bases, the characteristics of news events processing that may have contributed to its increased vulnerability relative to autobiographical memory (AbM). PMID- 15820670 TI - Malaria virulence genes controlling expression through chromatin modification. AB - Mutually exclusive expression within the var gene family of the malaria parasite Plasmodium falciparum is important for parasite survival and virulence. In this issue of Cell, Duraisingh et al. and Freitas-Junior et al. provide evidence for the role of Sir2-dependent alterations in chromatin structure and changes in subnuclear chromatin localization in regulating var gene expression (Duraisingh et al., 2005; Freitas-Junior et al., 2005). PMID- 15820671 TI - Ubiquitin-mediated degradation a mechanism for fine-tuning TGF-beta signaling. AB - Effects of the cytokine TGF-beta can be dampened by E3 ubiquitin ligases that target specific Smads, the TGF-beta signal transducers, for proteolytic destruction. Two papers in this issue of Cell highlight the importance of this mechanism in regulating the in vivo effects of TGF-beta. The first paper identifies and characterizes a novel Smad4 ubiquitin ligase, and the second paper redefines the role of a previously identified Smad1 ubiquitin ligase, Smurf-1 (Dupont et al., 2005; Yamashita et al., 2005). PMID- 15820672 TI - Axon guidance: opposing EPHects in the growth cone. AB - Cells communicate with other cells via (trans) interaction between membrane linked ephrins and Eph receptors. In this issue of Cell, Pfaff and colleagues (Marquardt et al., 2005) demonstrate that coexpressed ephrin-As and Ephs do not interact in cis but rather segregate into separate membrane domains, from which they signal opposing effects during motor axon guidance. PMID- 15820673 TI - Remote control of fruit fly behavior. AB - Electrical stimulation of neurons in the central nervous system of awake, behaving animals offers the ultimate test to determine whether the activation of specific neurons is sufficient to elicit perception, motor activity, or other behaviors. In this issue of Cell, Lima and Miesenbock (Lima and Miesenbock, 2005) dump the stimulating electrode in favor of a new remote control system to excite specific neurons--light activation of transgenically supplied ion channels. PMID- 15820674 TI - Melanocyte stem cell maintenance and hair graying. AB - Hair graying is an obvious sign of human aging, yet little was known about its causes. Two recent papers provide compelling evidence that hair graying is due to incomplete melanocyte stem cell maintenance and identify Pax3 and Mitf as key molecules that help regulate the balance between melanocyte stem cell maintenance and differentiation. PMID- 15820675 TI - Heterochromatin silencing and locus repositioning linked to regulation of virulence genes in Plasmodium falciparum. AB - The malaria parasite Plasmodium falciparum undergoes antigenic variation to evade host immune responses through switching expression of variant surface proteins encoded by the var gene family. We demonstrate that both a subtelomeric transgene and var genes are subject to reversible gene silencing. Var gene silencing involves the SIR complex as gene disruption of PfSIR2 results in activation of this gene family. We also demonstrate that perinuclear gene activation involves chromatin alterations and repositioning into a location that may be permissive for transcription. Together, this implies that locus repositioning and heterochromatic silencing play important roles in the epigenetic regulation of virulence genes in P. falciparum. PMID- 15820676 TI - Telomeric heterochromatin propagation and histone acetylation control mutually exclusive expression of antigenic variation genes in malaria parasites. AB - Malaria parasites use antigenic variation to avoid immune clearance and increase the duration of infection in the human host. Variation at the surface of P. falciparum-infected erythrocytes is mediated by the differential control of a family of surface antigens encoded by var genes. Switching of var gene expression occurs in situ, mostly from telomere-associated loci, without detectable DNA alterations, suggesting that it is controlled by chromatin structure. We have identified chromatin modifications at telomeres that spread far into telomere proximal regions, including var gene loci (>50 kb). One type of modification is mediated by a protein homologous to yeast Sir2 called PfSir2, which forms a chromosomal gradient of heterochromatin structure and histone hypoacetylation. Upon activation of a specific telomere-associated var gene, PfSir2 is removed from the promoter region and acetylation of histone occurs. Our data demonstrate that mutually exclusive transcription of var genes is linked to the dynamic remodeling of chromatin. PMID- 15820677 TI - Sumoylation silences the plasma membrane leak K+ channel K2P1. AB - Reversible, covalent modification with small ubiquitin-related modifier proteins (SUMOs) is known to mediate nuclear import/export and activity of transcription factors. Here, the SUMO pathway is shown to operate at the plasma membrane to control ion channel function. SUMO-conjugating enzyme is seen to be resident in plasma membrane, to assemble with K2P1, and to modify K2P1 lysine 274. K2P1 had not previously shown function despite mRNA expression in heart, brain, and kidney and sequence features like other two-P loop K+ leak (K2P) pores that control activity of excitable cells. Removal of the peptide adduct by SUMO protease reveals K2P1 to be a K+-selective, pH-sensitive, openly rectifying channel regulated by reversible peptide linkage. PMID- 15820678 TI - Mechanism of cross-species prion transmission: an infectious conformation compatible with two highly divergent yeast prion proteins. AB - Efficiency of interspecies prion transmission decreases as the primary structures of the infectious proteins diverge. Yet, a single prion protein can misfold into multiple infectious conformations, and such differences in "strain conformation" also alter infection specificity. Here, we explored the relationship between prion strains and species barriers by creating distinct synthetic prion forms of the yeast prion protein Sup35. We identified a strain conformation of Sup35 that allows transmission from the S. cerevisiae (Sc) Sup35 to the highly divergent C. albicans (Ca) Sup35 both in vivo and in vitro. Remarkably, cross-species transmission leads to a novel Ca strain that in turn can infect the Sc protein. Structural studies reveal strain-specific conformational differences in regions of the prion domain that are involved in intermolecular contacts. Our findings support a model whereby strain conformation is the critical determinant of cross species prion transmission while primary structure affects transmission specificity by altering the spectrum of preferred amyloid conformations. PMID- 15820679 TI - Fibril conformation as the basis of species- and strain-dependent seeding specificity of mammalian prion amyloids. AB - Spongiform encephalopathies are believed to be transmitted by self-perpetuating conformational conversion of the prion protein. It was shown recently that fundamental aspects of mammalian prion propagation can be reproduced in vitro in a seeded fibrillization of the recombinant prion protein variant Y145Stop (PrP23 144). Here we demonstrate that PrP23-144 amyloids from different species adopt distinct secondary structures and morphologies, and that these structural differences are controlled by one or two residues in a critical region. These sequence-specific structural characteristics correlate strictly with the seeding specificity of amyloid fibrils. However, cross-seeding of PrP23-144 from one species with preformed fibrils from another species may overcome natural sequence based structural preferences, resulting in a new amyloid strain that inherits the secondary structure and morphology of the template. These data provide direct biophysical evidence that protein conformations are transmitted in PrP amyloid strains, establishing a foundation for a structural basis of mammalian prion transmission barriers. PMID- 15820680 TI - The biological and chemical basis for tissue-selective amyloid disease. AB - Factors controlling the onset and progression of extracellular amyloid diseases remain largely unknown. Central to disease etiology is the efficiency of the endoplasmic reticulum (ER) machinery that targets destabilized mutant proteins for degradation and the enhanced tendency of these variants to aggregate if secreted. We demonstrate that mammalian cells secrete numerous transthyretin (TTR) disease-associated variants with wild-type efficiency in spite of compromised folding energetics. Only the most highly destabilized TTR variants are subjected to ER-associated degradation (ERAD) and then only in certain tissues, providing insight into tissue selective amyloidosis. Rather than a "quality control" standard based on wild-type stability, we find that ER-assisted folding (ERAF), based on global protein energetics, determines the extent of export. We propose that ERAF (influenced by the energetics of the protein fold, chaperone enzyme distributions, and metabolite chaperones) in competition with ERAD defines the unique secretory aptitude of each tissue. PMID- 15820681 TI - Germ-layer specification and control of cell growth by Ectodermin, a Smad4 ubiquitin ligase. AB - TGF-beta signaling is essential for development and proliferative homeostasis. During embryogenesis, maternal determinants act in concert with TGF-beta signals to form mesoderm and endoderm. In contrast, ectoderm specification requires the TGF-beta response to be attenuated, although the mechanisms by which this is achieved remain unknown. In a functional screen for ectoderm determinants, we have identified Ectodermin (Ecto). In Xenopus embryos, Ecto is essential for the specification of the ectoderm and acts by restricting the mesoderm-inducing activity of TGF-beta signals to the mesoderm and favoring neural induction. Ecto is a RING-type ubiquitin ligase for Smad4, a TGF-beta signal transducer. Depletion of Ecto in human cells enforces TGF-beta-induced cytostasis and, moreover, plays a causal role in limiting the antimitogenic effects of Smad4 in tumor cells. We propose that Ectodermin is a key switch in the control of TGF beta gene responses during early embryonic development and cell proliferation. PMID- 15820682 TI - Ubiquitin ligase Smurf1 controls osteoblast activity and bone homeostasis by targeting MEKK2 for degradation. AB - Bone is constantly resorbed and formed throughout life by coordinated actions of osteoclasts and osteoblasts. Here we show that Smurf1, a HECT domain ubiquitin ligase, has a specific physiological role in suppressing the osteogenic activity of osteoblasts. Smurf1-deficient mice are born normal but exhibit an age dependent increase of bone mass. The cause of this increase can be traced to enhanced activities of osteoblasts, which become sensitized to bone morphogenesis protein (BMP) in the absence of Smurf1. However, loss of Smurf1 does not affect the canonical Smad-mediated intracellular TGFbeta or BMP signaling; instead, it leads to accumulation of phosphorylated MEKK2 and activation of the downstream JNK signaling cascade. We demonstrate that Smurf1 physically interacts with MEKK2 and promotes the ubiquitination and turnover of MEKK2. These results indicate that Smurf1 negatively regulates osteoblast activity and response to BMP through controlling MEKK2 degradation. PMID- 15820683 TI - JNK extends life span and limits growth by antagonizing cellular and organism wide responses to insulin signaling. AB - Aging of a eukaryotic organism is affected by its nutrition state and by its ability to prevent or repair oxidative damage. Consequently, signal transduction systems that control metabolism and oxidative stress responses influence life span. When nutrients are abundant, the insulin/IGF signaling (IIS) pathway promotes growth and energy storage but shortens life span. The transcription factor Foxo, which is inhibited by IIS, extends life span in conditions of low IIS activity. Life span can also be increased by activating the stress-responsive Jun-N-terminal kinase (JNK) pathway. Here we show that JNK requires Foxo to extend life span in Drosophila. JNK antagonizes IIS, causing nuclear localization of Foxo and inducing its targets, including growth control and stress defense genes. JNK and Foxo also restrict IIS activity systemically by repressing IIS ligand expression in neuroendocrine cells. The convergence of JNK signaling and IIS on Foxo provides a model to explain the effects of stress and nutrition on longevity. PMID- 15820684 TI - Coexpressed EphA receptors and ephrin-A ligands mediate opposing actions on growth cone navigation from distinct membrane domains. AB - Contact-dependent signaling between membrane-linked ligands and receptors such as the ephrins and Eph receptor tyrosine kinases controls a wide range of developmental and pathological processes. Paradoxically, many cell types coexpress both ligands and receptors, raising the question of how specific signaling readouts are achieved under these conditions. Here, we studied the signaling activities exerted by coexpressed EphA receptors and GPI-linked ephrin A ligands in spinal motor neuron growth cones. We demonstrate that coexpressed Eph and ephrin proteins segregate laterally into distinct membrane domains from which they signal opposing effects on the growth cone: EphAs direct growth cone collapse/repulsion and ephrin-As signal motor axon growth/attraction. This subcellular arrangement of Eph-ephrin proteins enables axons to discriminate between cis- versus trans-configurations of ligand/receptor proteins, thereby allowing the utilization of both Ephs and ephrins as functional guidance receptors within the same neuronal growth cone. PMID- 15820685 TI - Remote control of behavior through genetically targeted photostimulation of neurons. AB - Optically gated ion channels were expressed in circumscribed groups of neurons in the Drosophila CNS so that broad illumination of flies evoked action potentials only in genetically designated target cells. Flies harboring the "phototriggers" in different sets of neurons responded to laser light with behaviors specific to the sites of phototrigger expression. Photostimulation of neurons in the giant fiber system elicited the characteristic escape behaviors of jumping, wing beating, and flight; photostimulation of dopaminergic neurons caused changes in locomotor activity and locomotor patterns. These responses reflected the direct optical activation of central neuronal targets rather than confounding visual input, as they persisted unabated in carriers of a mutation that eliminates phototransduction. Encodable phototriggers provide noninvasive control interfaces for studying the connectivity and dynamics of neural circuits, for assigning behavioral content to neurons and their activity patterns, and, potentially, for restoring information corrupted by injury or disease. PMID- 15820686 TI - Antidepressants and the monoamine masquerade. AB - Neurotransmitter transporters have long been known to recognize related compounds as substrates, resulting in the accumulation and release of so-called "false transmitters." In this issue of Neuron, Zhou et al. show that when serotonin levels are elevated by inhibition of either serotonin reuptake or of monoamine oxidase, dopamine neurons accumulate serotonin. The results suggest that release of serotonin by dopamine neurons may contribute to the effects of multiple major classes of antidepressants. PMID- 15820687 TI - CAPS in search of a lost function. AB - Ca2+-dependent activator protein for secretion (CAPS) is an evolutionarily conserved secretory protein that was previously thought to mediate Ca2+-triggered fusion of dense-core vesicles. In an elegant study of CAPS1-deficient mice, Speidel et al. (this issue of Neuron) now show that CAPS function may have been misunderstood. CAPS appears to act upstream of fusion in the biogenesis or maintenance of mature secretory vesicles, raising the possibility of a completely new type of function for an essential component of the secretory machinery. PMID- 15820688 TI - Timing and prediction the code from basal ganglia to thalamus. AB - When is an inhibitory synapse not inhibitory? In this issue of Neuron, Person and Perkel demonstrate that thalamic neurons can translate extrinsic GABAergic input from the basal ganglia into highly precise patterns of sustained spiking in a circuit that is essential for vocal learning in songbirds. Postinhibitory rebound serves as a mechanism that preserves precise spike timing information, enabling reliable propagation of activity throughout this pathway. The results have broad implications for basic mechanisms of functional processing in both thalamus and basal ganglia and serve to increase our understanding of how acoustic units of vocal sounds are transformed into motor gestures during the sensitive period for song learning. PMID- 15820689 TI - Pain TRPs. AB - Transient receptor potential (TRP) ion channels are molecular gateways in sensory systems, an interface between the environment and the nervous system. Several TRPs transduce thermal, chemical, and mechanical stimuli into inward currents, an essential first step for eliciting thermal and pain sensations. Precise regulation of the expression, localization, and function of the TRP channels is crucial for their sensory role in nociceptor terminals, particularly after inflammation, when they contribute to pain hypersensitivity by undergoing changes in translation and trafficking as well as diverse posttranslational modifications. PMID- 15820690 TI - A chemical-genetic approach to studying neurotrophin signaling. AB - Trk tyrosine kinases are receptors for members of the neurotrophin family and are crucial for growth and survival of specific populations of neurons. Yet, the functions of neurotrophin-Trk signaling in postnatal development as well as maintenance and plasticity of the adult nervous system are less clear. We report here the generation of mice harboring Trk knockin alleles that allow for pharmacological control of Trk kinase activity. Nanomolar concentrations of either 1NMPP1 or 1NaPP1, derivatives of the general kinase inhibitor PP1, inhibit NGF and BDNF signaling in TrkA(F592A) and TrkB(F616A) neurons, respectively, while no such Trk inhibition is observed in wild-type neurons. Moreover, oral administration of 1NMPP1 leads to specific inhibition of TrkA(F592A), TrkB(F616A), and TrkC(F167A) signaling in vivo. Thus, Trk knockin mice provide valuable tools for selective, rapid, and reversible inhibition of neurotrophin signaling in vitro and in vivo. PMID- 15820691 TI - Frizzled 5 signaling governs the neural potential of progenitors in the developing Xenopus retina. AB - Progenitors in the developing central nervous system acquire neural potential and proliferate to expand the pool of precursors competent to undergo neuronal differentiation. The formation and maintenance of neural-competent precursors are regulated by SoxB1 transcription factors, and evidence that their expression is regionally regulated suggests that specific signals regulate neural potential in subdomains of the developing nervous system. We show that the frizzled (Fz) transmembrane receptor Xfz5 selectively governs neural potential in the developing Xenopus retina by regulating the expression of Sox2. Blocking either Xfz5 or canonical Wnt signaling within the developing retina inhibits Sox2 expression, reduces cell proliferation, inhibits the onset of proneural gene expression, and biases individual progenitors toward a nonneural fate, without altering the expression of multiple progenitor markers. Blocking Sox2 function mimics these effects. Rescue experiments indicate that Sox2 is downstream of Xfz5. Thus, Fz signaling can regulate the neural potential of progenitors in the developing nervous system. PMID- 15820692 TI - Cholinergic input is required during embryonic development to mediate proper assembly of spinal locomotor circuits. AB - Rhythmic limb movements are controlled by pattern-generating neurons within the ventral spinal cord, but little is known about how these locomotor circuits are assembled during development. At early stages of embryogenesis, motor neurons are spontaneously active, releasing acetylcholine that triggers the depolarization of adjacent cells in the spinal cord. To investigate whether acetylcholine-driven activity is required for assembly of the central pattern-generating (CPG) circuit, we studied mice lacking the choline acetyltransferase (ChAT) enzyme. Our studies show that a rhythmically active spinal circuit forms in ChAT mutants, but the duration of each cycle period is elongated, and right-left and flexor extensor coordination are abnormal. In contrast, blocking acetylcholine receptors after the locomotor network is wired does not affect right-left or flexor extensor coordination. These findings suggest that the cholinergic neurotransmitter pathway is involved in configuring the CPG during a transient period of development. PMID- 15820693 TI - LIN-23-mediated degradation of beta-catenin regulates the abundance of GLR-1 glutamate receptors in the ventral nerve cord of C. elegans. AB - Ubiquitin-mediated protein degradation has been proposed to play an important role in regulating synaptic transmission. Here we show that LIN-23, the substrate binding subunit of a Skp1/Cullin/F Box (SCF) ubiquitin ligase, regulates the abundance of the glutamate receptor GLR-1 in the ventral nerve cord of C. elegans. Mutants lacking lin-23 had an increased abundance of GLR-1 in the ventral cord. The increase of GLR-1 was not caused by changes in the ubiquitination of GLR-1. Instead, SCF(LIN-23) regulates GLR-1 through the beta catenin homolog BAR-1 and the TCF/Lef transcription factor homolog POP-1. We hypothesize that LIN-23-mediated degradation of BAR-1 beta-catenin regulates the transcription of Wnt target genes, which in turn alter postsynaptic properties. PMID- 15820694 TI - Corelease of dopamine and serotonin from striatal dopamine terminals. AB - The striatum receives rich dopaminergic and more moderate serotonergic innervation. After vesicular release, dopamine and serotonin (5 hydroxytryptamine, 5-HT) signaling is controlled by transporter-mediated reuptake. Dopamine is taken up by dopamine transporters (DATs), which are expressed at the highest density in the striatum. Although DATs also display a low affinity for 5-HT, that neurotransmitter is normally efficiently taken up by the 5-HT transporters. We found that when extracellular 5-HT is elevated by exogenous application or by using antidepressants (e.g., fluoxetine) to inhibit the 5-HT transporters, the extremely dense striatal DATs uptake 5-HT into dopamine terminals. Immunohistochemical results and measurements using fast cyclic voltammetry showed that elevated 5-HT is taken up by DATs into striatal dopamine terminals that subsequently release 5-HT and dopamine together. These results suggest that antidepressants that block serotonin transporters or other factors that elevate extracellular 5-HT alter the temporal and spatial relationship between dopamine and 5-HT signaling in the striatum. PMID- 15820695 TI - CAPS1 regulates catecholamine loading of large dense-core vesicles. AB - CAPS1 is thought to play an essential role in mediating exocytosis from large dense-core vesicles (LDCVs). We generated CAPS1-deficient (KO) mice and studied exocytosis in a model system for Ca2+-dependent LDCV secretion, the adrenal chromaffin cell. Adult heterozygous CAPS1 KO cells display a gene dosage dependent decrease of CAPS1 expression and a concomitant reduction in the number of docked vesicles and secretion. Embryonic homozygous CAPS1 KO cells show a strong reduction in the frequency of amperometrically detectable release events of transmitter-filled vesicles, while the total number of fusing vesicles, as judged by capacitance recordings or total internal reflection microscopy, remains unchanged. We conclude that CAPS1 is required for an essential step in the uptake or storage of catecholamines in LDCVs. PMID- 15820696 TI - mGluR7 is a metaplastic switch controlling bidirectional plasticity of feedforward inhibition. AB - Plasticity of feedforward inhibition in the hippocampal mossy fiber (MF) pathway can dramatically influence dentate gyrus-CA3 dialog. Interestingly, MF inputs to CA3 stratum lucidum interneurons (SLINs) undergo long-term depression (LTD) following high-frequency stimulation (HFS), in contrast to MF-pyramid (PYR) synapses, where long-term potentiation (LTP) occurs. Furthermore, activity induced potentiation of MF-SLIN transmission has not previously been observed. Here we report that metabotropic glutamate receptor subtype 7 (mGluR7) is a metaplastic switch at MF-SLIN synapses, whose activation and surface expression governs the direction of plasticity. In naive slices, mGluR7 activation during HFS generates MF-SLIN LTD, depressing presynaptic release through a PKC-dependent mechanism. Following agonist exposure, mGluR7 undergoes internalization, unmasking the ability of MF-SLIN synapses to undergo presynaptic potentiation in response to the same HFS that induces LTD in naive slices. Thus, selective mGluR7 targeting to MF terminals contacting SLINs and not PYRs provides cell target specific plasticity and bidirectional control of feedforward inhibition. PMID- 15820697 TI - Activity-dependent adjustments of the inhibitory network in the olfactory bulb following early postnatal deprivation. AB - The first-order sensory relay for olfactory processing, the main olfactory bulb (MOB), retains the ability to acquire new interneurons throughout life. It is therefore a particularly appropriate region for studying the role of experience in sculpting neuronal networks. We found that nostril closure decreased the number of newborn granule cells in the MOB, the complexity of their dendritic arborization, and their spine density, without affecting the preexisting population of granule cells. Accordingly, the frequency of miniature synaptic inhibitory events received by mitral cells was reduced. However, due to a compensatory increase in newborn granule cell excitability, action potential dependent GABA release was dramatically enhanced, thus counteracting the reduction in spine density and leading to an unaltered synchronization of mitral cell firing activity. Together, this study reveals a unique form of adaptive response brought about exclusively by the cohort of newborn cells and used to maintain normal functioning of the MOB. PMID- 15820698 TI - Directional selectivity is formed at multiple levels by laterally offset inhibition in the rabbit retina. AB - The excitatory and inhibitory inputs to directionally selective (DS) ganglion cells are themselves directionally selective. Directionality is achieved because excitation is reduced during null-direction movement along a GABAergic pathway. Inhibition is reduced during preferred-direction movement along a pathway that includes cholinergic synapses. Both excitation and inhibition are made directional by laterally offset inhibitory signals similar to the spatial offset of the direct inhibitory input to the DS cell dendrites. Thus, spatially offset lateral inhibition generates directionality at three different levels in the DS circuitry. We also found that for stimuli falling within the dendritic field, cholinergic input is delivered to the OFF but not the ON dendrites. Cholinergic pathways from outside the dendritic field reach both ON and OFF dendrites, but both of these pathways are normally inactivated by GABAergic synapses. PMID- 15820699 TI - Unitary IPSPs drive precise thalamic spiking in a circuit required for learning. AB - Song learning in birds requires a basal ganglia-thalamo-pallial loop that contains a calyceal GABAergic synapse in the thalamus. Information processing within this circuit is critical for proper song development; however, it is unclear whether activation of the inhibitory output of the basal ganglia structure Area X can drive sustained activity in its thalamic target, the medial portion of the dorsolateral thalamic nucleus (DLM). We show that high-frequency, random activation of this GABAergic synapse can drive precisely timed firing in DLM neurons in brain slices in the absence of excitatory input. Complex IPSP trains, including spike trains recorded in vivo, drive spiking in slices with high reproducibility, even between animals. Using a simple model, we can predict much of DLM's response to natural stimulus trains. These data elucidate basic rules by which thalamic relay neurons translate IPSPs into suprathreshold output and demonstrate extrathalamic GABAergic activation of thalamus. PMID- 15820700 TI - Prefrontal phase locking to hippocampal theta oscillations. AB - The interactions between cortical and hippocampal circuits are critical for memory formation, yet their basic organization at the neuronal network level is not well understood. Here, we demonstrate that a significant portion of neurons in the medial prefrontal cortex of freely behaving rats are phase locked to the hippocampal theta rhythm. In addition, we show that prefrontal neurons phase lock best to theta oscillations delayed by approximately 50 ms and confirm this hippocampo-prefrontal directionality and timing at the level of correlations between single cells. Finally, we find that phase locking of prefrontal cells is predicted by the presence of significant correlations with hippocampal cells at positive delays up to 150 ms. The theta-entrained activity across cortico hippocampal circuits described here may be important for gating information flow and guiding the plastic changes that are believed to underlie the storage of information across these networks. PMID- 15820701 TI - Motor learning by observing. AB - Learning complex motor behaviors like riding a bicycle or swinging a golf club is based on acquiring neural representations of the mechanical requirements of movement (e.g., coordinating muscle forces to control the club). Here we provide evidence that mechanisms matching observation and action facilitate motor learning. Subjects who observed a video depicting another person learning to reach in a novel mechanical environment (imposed by a robot arm) performed better when later tested in the same environment than subjects who observed similar movements but no learning; moreover, subjects who observed learning of a different environment performed worse. We show that this effect is not based on conscious strategies but instead depends on the implicit engagement of neural systems for movement planning and control. PMID- 15820702 TI - Memory of early maltreatment: neonatal behavioral and neural correlates of maternal maltreatment within the context of classical conditioning. AB - BACKGROUND: While children form an attachment to their abusive caregiver, they are susceptible to mental illness and brain abnormalities. To understand this important clinical issue, we have developed a rat animal model of abusive attachment where odor paired with shock paradoxically produces an odor preference. Here, we extend this model to a seminaturalistic paradigm using a stressed, "abusive" mother during an odor presentation and assess the underlying learning neural circuit. METHODS: We used a classical conditioning paradigm pairing a novel odor with a stressed mother that predominantly abused pups to assess olfactory learning in a seminaturalistic environment. Additionally, we used Fos protein immunohistochemistry to assess brain areas involved in learning this pain-induced odor preference within a more controlled maltreatment environment (odor-shock conditioning). RESULTS: Odor-maternal maltreatment pairings within a seminatural setting and odor-shock pairings both resulted in paradoxical odor preferences. Learning-induced gene expression was altered in the olfactory bulb and anterior piriform cortex (part of olfactory cortex) but not the amygdala. CONCLUSIONS: Infants appear to use a unique brain circuit that optimizes learned odor preferences necessary for attachment. A fuller understanding of infant brain function may provide insight into why early maltreatment affects psychiatric well-being. PMID- 15820703 TI - Corticolimbic blood flow in posttraumatic stress disorder during script-driven imagery. AB - BACKGROUND: Functional neuroimaging experiments targeting personal recall of emotional events may help elucidate neural substrates underlying posttraumatic stress disorder (PTSD). Studies suggest that limbic and paralimbic function might be altered in PTSD, as compared with trauma-exposed control subjects; however, little is known about functional changes resulting from traumatic experience itself. The present study examined both PTSD-specific and trauma-specific regional cerebral blood flow (rCBF) patterns during script-driven imagery. METHODS: Sixteen combat veterans with PTSD (PP); 15 combat veterans without PTSD (CC); and 14 healthy, aged-matched noncombat control subjects (NC) underwent [15O] H20 positron emission tomography (PET) scanning during script-driven imagery of emotionally evocative and neutral autobiographic events. RESULTS: Differential patterns of activation were detected in amygdala and medial frontal cortex. Past trauma experience was associated with decreased amygdala activity (i.e., less activity than healthy control subjects); however, combat control subjects deactivated this region (i.e., greater activity to neutral scripts). All subjects deactivated medial frontal cortex; PTSD patients had greater rostral anterior cingulate (rACC) deactivation compared with control groups, who deactivated ventromedial prefrontal cortex (vmPFC). CONCLUSIONS: Trauma-specific patterns may represent potential compensatory changes to traumatic reminders, while patterns observed only in the PTSD group may reflect neural substrates specific to PTSD pathophysiology. PMID- 15820704 TI - D-cycloserine facilitates extinction of learned fear: effects on reacquisition and generalized extinction. AB - BACKGROUND: d-cycloserine (DCS) facilitates extinction of learned fear. The aim of this study was to examine whether DCS 1) affects reacquisition of fear (Experiment 1) and 2) produces generalized extinction of fear (Experiment 2). METHODS: Following fear conditioning, where a light or a tone conditioned stimulus (CS) was paired with a white-noise burst unconditioned stimulus (US), rats received nonreinforced exposure to one CS (i.e., extinction training). Fear was assessed by measuring CS-elicited freezing, a species-specific defense response. RESULTS: Rats given DCS exhibited facilitated extinction of fear but were able to reacquire fear of that CS in a similar manner as saline-treated control animals (Experiment 1). Furthermore, DCS-treated rats exhibited generalized extinction (i.e., they were less fearful of a non-extinguished CS) in comparison to controls (Experiment 2). CONCLUSIONS: DCS facilitates extinction of learned fear to the extinguished CS, but also appears to reduce fear of a nonextinguished CS. These findings suggest that this drug may have substantial clinical value in the treatment of anxiety disorders. PMID- 15820705 TI - Mild early life stress enhances prefrontal-dependent response inhibition in monkeys. AB - BACKGROUND: Severely stressful early experiences have been implicated in the pathophysiology of psychiatric disorders. In contrast, exposure to mild early life stress (i.e., stress inoculation) strengthens emotional and neuroendocrine resistance to subsequent stressors. Herein we extend this research to examine the effects of mild early life stress on cognition. METHODS: Squirrel monkeys were randomized to a mild intermittent stress (IS; n = 11) or nonstress (NS; n = 9) condition from 17 to 27 weeks postpartum. At 1.5 years of age, monkeys were assessed for response inhibition on a test previously shown to reflect prefrontal dependent cognitive function. RESULTS: IS monkeys demonstrated fewer response inhibition errors compared with NS monkeys. There were no rearing-related differences in aspects of performance that did not require inhibitory control. Compared with NS monkeys, IS monkeys had lower basal plasma pituitary-adrenal stress hormone levels. No rearing-related differences on neuroendocrine measures obtained 15 minutes after testing were found. CONCLUSIONS: Results from this experiment provide the first evidence that exposure to mildly stressful early experiences improves prefrontal-dependent response inhibition in primates. Combined with our previous data, findings from this animal model suggest that exposure to mild early life stress may enhance the development of brain systems that regulate emotional, neuroendocrine, and cognitive control. PMID- 15820706 TI - Acute stress-induced impairment of spatial memory is associated with decreased expression of neural cell adhesion molecule in the hippocampus and prefrontal cortex. AB - BACKGROUND: There is an extensive literature describing how stress disturbs cognitive processing and can exacerbate psychiatric disorders. There is, however, an insufficient understanding of the molecular mechanisms involved in stress effects on brain and behavior. METHODS: Rats were given spatial memory training in a hippocampus-dependent water maze task. We investigated how a fear-provoking experience (predator exposure) would affect their spatial memory and neural cell adhesion molecule (NCAM) levels in the hippocampus, prefrontal cortex (PFC), amygdala, and cerebellum. RESULTS: Whereas the control (nonstress) group exhibited excellent memory for the hidden platform location in the water maze, the cat-exposed (stress) group exhibited a profound impairment of memory and a marked suppression of levels of the NCAM-180 isoform in the hippocampus. Predator stress produced a more global reduction of NCAM levels in the PFC but had no effect on NCAM levels in the amygdala and cerebellum. CONCLUSIONS: This work provides a novel perspective into dynamic and structure-specific changes in the molecular events involved in learning, memory, and stress. The selective suppression of NCAM-180 in the hippocampus and the more general suppression of NCAM in the PFC provide insight into the mechanisms underlying the great sensitivity of these two structures to be disturbed by stress. PMID- 15820707 TI - A stress-induced anxious state in male rats: corticotropin-releasing hormone induces persistent changes in associative learning and startle reactivity. AB - BACKGROUND: Exposure to intense inescapable stressors induces a persistent anxious state in rats. The anxious state is evident as increased sensory reactivity and enhanced associative learning. METHODS: We examine whether similar neurobehavioral changes are observed after intracerebroventricular (ICV) administration of corticotropin releasing hormone (CRH). Two behaviors were observed: acoustic startle responses (ASRs) and acquisition of the classically conditioned eyeblink response. Male Sprague-Dawley rats were administered ICV CRH either in a single dose (1.0 microg/rat) or in three doses each separated by 30 min. RESULTS: Exaggerated ASRs were evident 2 hours after either CRH treatment; however, only the rats given three injections exhibited a persistently exaggerated ASR apparent 24 hours after CRH treatment. Rats administered three injections of CRH also exhibited faster acquisition of the eyeblink conditioned response beginning 24 hours after treatment. Yet, we did not find evidence for a persistent activation of the HPA-axis response; three CRH injections did not lead to elevated basal plasma corticosterone levels the following morning. CONCLUSIONS: Repeated treatment with CRH over a 1.5-hour period models some of the behavioral changes observed after exposure to intense inescapable stressors. PMID- 15820708 TI - Functional connectivity of dissociative responses in posttraumatic stress disorder: a functional magnetic resonance imaging investigation. AB - BACKGROUND: The purpose of this study was to assess interregional brain activity covariations during traumatic script-driven imagery in subjects with posttraumatic stress disorder (PTSD). METHODS: Functional magnetic resonance imaging and functional connectivity analyses were used to assess interregional brain activity covariations during script-driven imagery in PTSD subjects with a dissociative response, PTSD subjects with a flashback response, and healthy control subjects. RESULTS: Significant between-group differences in functional connectivity were found. Comparing dissociated PTSD patients and control subjects' connectivity maps in the left ventrolateral thalamus (VLT) [-14, -16, 4] revealed that control subjects had higher covariations between activations in VLT and in the left superior frontal gyrus (Brodmann's area [BA] 10), right parahippocampal gyrus (BA 30), and right superior occipital gyrus (BA 19, 39), whereas greater covariation with VLT in dissociated PTSD subjects occurred in the right insula (BA 13, 34), left parietal lobe (BA 7), right middle frontal gyrus (BA 8), superior temporal gyrus (BA 38, 34), and right cuneus (BA 19). Comparing dissociated PTSD and flashback PTSD connectivity maps in the right cingulate gyrus [3, 16, 30] revealed that dissociated PTSD subjects had higher covariations between activations in this region and the left inferior frontal gyrus (BA 47). CONCLUSIONS: Greater activation of neural networks involved in representing bodily states was seen in dissociated PTSD subjects than in non-PTSD control subjects. These findings might illuminate the mechanisms underlying distorted body perceptions often observed clinically during dissociative episodes. PMID- 15820709 TI - Peroxisome proliferator-activated receptor gamma activation decreases neuroinflammation in brain after stress in rats. AB - BACKGROUND: A growing body of evidence has demonstrated that peroxisome proliferator-activated receptor gamma (PPARgamma) play a role in brain inflammatory conditions because various PPARgamma ligands inhibit proinflammatory mediators, such as cytokines (tumor necrosis factor alpha [TNFalpha]) and inducible nitric oxide synthase (NOS-2). As has been previously shown, immobilization stress and stress-related neuropsychologic conditions are followed by accumulation of oxidative/nitrosative mediators in brain after the release of cytokines, nuclear factor kappaB activation, and NOS-2 and cyclooxygenase 2 (COX 2) expression in the brain. METHODS: To assess whether PPARgamma activation can modify the accumulation of oxidative/nitrosative species seen in brain after stress, and to study the mechanisms by which this effect is achieved, young-adult male Wistar rats (control and immobilized during 6 hours) were injected (IP) with the high-affinity ligand rosiglitazone (RS) at the onset of stress. RESULTS: Stress increased PPARgamma expression in cortical neurons and glia as assessed by Western blot and immunohistochemistry. In stressed animals, RS (1-3 mg/kg) decreased stress-induced increases in NOS-2 activity. On the other hand, the PPARgamma ligand decreased stress-induced malondialdehyde (an indicator of lipid peroxidation) accumulation in cortex and prevented oxidation of the main antioxidant glutathione. The mechanisms involved in the antioxidative properties of RS in stress involve nuclear factor KB blockade (by preventing stress-induced IkappaBalpha decrease) and inhibition of TNFalpha release in stressed animals. At the doses tested, RS did not decrease COX-2 expression and prostaglandin E2 release during stress. Finally, RS also decreased chronic (repeated immobilization for 21 days) stress-induced accumulation of oxidative/nitrosative mediators. CONCLUSIONS: Taken together, these findings suggest a role for this antiinflammatory pathway in the brain response to stress and the possibility of pharmacologic modulation for preventing accumulation of oxidative/nitrosative species and subsequent brain damage in stress-related neuropsychologic conditions. PMID- 15820710 TI - Familial and sporadic subtypes of early-onset Obsessive-Compulsive disorder. AB - BACKGROUND: Family studies of Obsessive-Compulsive (OCD) indicate there is substantial heterogeneity in the familiality of the disorder. This study was done to determine whether there are differences between familial and sporadic probands with early-onset OCD in obsessive-compulsive (OC) symptom categories and comorbid psychiatric diagnoses. METHODS: We ascertained 50 OCD probands ranging in age from 10 to 19 years with an onset of OC symptoms before age 15 years. All probands were directly assessed with semistructured diagnostic interviews; their first-degree and second-degree relatives were directly or indirectly assessed with similar diagnostic instruments. Descriptive data were compared in 33 familial and 17 sporadic OCD probands using logistic regression to control for age, gender, and age at onset of OC symptoms. RESULTS: Ordering compulsions were significantly more common in the familial OCD probands. Aberrant grooming behaviors were significantly more frequent in the familial subgroup with skin picking contributing significantly to that difference. Anxiety disorders other than OCD were also significantly more frequent in the familial subgroup with phobic disorders contributing significantly to that difference. CONCLUSIONS: The results indicate that familial and sporadic forms of early-onset OCD may be differentiated by ordering compulsions, aberrant grooming behaviors, and anxiety disorders other than OCD. PMID- 15820711 TI - Brain activation of patients with obsessive-compulsive disorder during neuropsychological and symptom provocation tasks before and after symptom improvement: a functional magnetic resonance imaging study. AB - BACKGROUND: Functional neuroimaging studies have implicated hyperactivity of the frontal cortex in obsessive-compulsive disorder (OCD); however, relationships between abnormal brain activity, clinical improvement, and neuropsychological function have not been clarified in OCD. To clarify the pathophysiology of this disorder, regional changes in brain function were examined during administration of cognitive and symptom provocation tasks in patients with OCD before and after treatment. METHODS: Ten outpatients with OCD participated in the study. Functional magnetic resonance imaging (fMRI) was performed before and after treatment. Stroop and symptom provocation tasks were administered during fMRI. Each patient was randomly allocated to receive either pharmacotherapy with fluvoxamine 200 mg/day (n = 4) or behavior therapy (n = 6) for 12 weeks. RESULTS: After 12-week treatment, mean (+/- SD) total score on the Yale-Brown Obsessive Compulsive Scale decreased from 29.00 +/- 3.59 to 14.60 +/- 9.22, representing symptomatic improvement from moderate to mild. After symptom improvement, symptom provocation-related activation in the orbitofrontal, dorsolateral-prefrontal, and anterior cingulate cortices decreased. Conversely, Stroop task-related activation in the parietal cortex and cerebellum increased. CONCLUSIONS: After improvement of OCD with either fluvoxamine or behavioral therapy, hyperactivation of the frontal lobe related to a symptom-provocative state decreases, and posterior brain activity related to action-monitoring function increases. PMID- 15820712 TI - The role of stress-induced cortisol in the relationship between depression and decreased bone mineral density. AB - BACKGROUND: This study was designed to test the hypothesis that cortisol mediates the relationship between bone density and depression in postmenopausal women. METHODS: Nineteen women aged 52-79 who had been assessed for bone mineral density by dual-energy x-ray absorptiometer (DEXA) were evaluated for depression and anxiety. Diurnal and stress-induced measures of salivary cortisol were obtained during the following week and at a laboratory session involving a speech task. RESULTS: Nine volunteers reported depression while 10 were never depressed. Ever depressed women had significantly lower total lumbar and right femur DEXA Z scores than never depressed (t(17) = 2.5, p = .019 and t(17) = 2.06, p = .05, respectively). Ever depressed women demonstrated a significant increase in salivary cortisol (area under the curve (AUC) = 27.83, SD = 37.64) compared to never depressed women (AUC = -13.34, SD = 19.55) (t(17) = -3.041, p = .007) during a psychological challenge. There were significant inverse relationships between salivary cortisol AUC values and bone density Z scores at every measured bone site. Mediation analyses suggest that 51 - 67% of the association between depression and bone density could be attributed to stress-induced changes in cortisol. CONCLUSIONS: Cortisol hypersecretion in response to stress may, in part, explain the impact of depression on bone density in post-menopausal women. PMID- 15820713 TI - Frontocortical 5-HT4 receptors exert positive feedback on serotonergic activity: viral transfections, subacute and chronic treatments with 5-HT4 agonists. AB - BACKGROUND: We recently identified a facilitory control exerted by serotonin4 (5 HT4) receptors on the in vivo firing activity of dorsal raphe nucleus (DRN) serotonergic (5-HT) neurons. However, these findings were based on acute administrations of 5-HT4 receptor agonists and antagonists, which were active only in a subpopulation of 5-HT neurons. We had no evidence that this influence was significant when considering the entire DRN, nor if it was persistent after chronic treatments. In addition, the poor distribution of 5-HT4 receptors within the DRN raised the question of the neuroanatomical bases underlying this control. METHODS AND RESULTS: Here we show that the subacute intraperitoneal (IP) injection of the 5-HT4 receptor agonists prucalopride (2.5 mg/kg) and RS 67333 (1.5 mg/kg) 30 minutes before the beginning of recordings augment the mean firing rate of DRN neurons by 40% and 66%, respectively. These increases remain stable when the compounds are administered continuously during 3 and 21 days; the effects of the 3-day treatment are blocked by the 5-HT4 receptor antagonist GR 125487 (1000 microg/kg, intravenous [i.v.]). In addition, stereotaxic microinjections of herpes simplex viruses, transformed to overexpress 5-HT4 receptors, increase DRN 5-HT neuronal mean activity when performed in the medial prefrontal cortex (mPFC) but not in the striatum or in the hippocampus. CONCLUSIONS: This finding suggests the existence of a 5-HT(4)-dependent activation of DRN that may involve the mPFC, unveiling the 5-HT4 receptor as a putative player in the physiopathology of several disorders related to central 5 HT dysfunction. PMID- 15820714 TI - Investigation of peptide YY and ghrelin responses to a test meal in bulimia nervosa. AB - BACKGROUND: Gut-derived peptides, such as peptide YY (PYY) and ghrelin that regulate the initiation and termination of meals, could play a role in the altered eating behavior of patients with bulimia nervosa (BN). Therefore, we aimed to assess plasma PYY and ghrelin responses to a test meal in symptomatic bulimics. METHODS: Ten healthy women and nine women with BN underwent blood sample collections before and after the ingestion of a test meal of 1300 Kcal (with 15% carbohydrates, 10% proteins, and 75% fat) at 12:00 noon. Plasma total PYY, ghrelin, insulin, and glucose were assayed. RESULTS: As compared with healthy women, bulimics exhibited a significantly blunted increase of circulating PYY (p < .007) and a significantly reduced suppression of plasma ghrelin (p < .0004) after the test meal. No significant differences emerged in food-induced plasma insulin and glucose changes between the two groups. Plasma ghrelin suppression after the meal was significantly correlated with plasma PYY increase. CONCLUSIONS: We replicated our previous findings of an altered ghrelin response to food ingestion in people with BN and showed for the first time a blunted PYY increase after food consumption in these patients. These findings support the occurrence in BN of a profound dysregulation of some peripheral regulatory mechanisms involved in the short-term regulation of feeding behavior that might be involved in the pathophysiology of their binge eating behavior. PMID- 15820715 TI - Analysis of pyramidal neuron morphology in an inducible knockout of brain-derived neurotrophic factor. AB - BACKGROUND: The messenger ribonucleic acid (mRNA) for brain-derived neurotrophic factor (BDNF), a regulator of pyramidal neuron dendritic spine density during development, is decreased in the prefrontal cortex of subjects with schizophrenia, and the level of BDNF mRNA expression is positively correlated with dendritic spine density in the same subjects. METHODS: To determine whether reduced BDNF mRNA expression might account for decreased spine density in schizophrenia, a knockout of the BDNF gene was induced in mice during embryogenesis or at 12 weeks of age. Quantitative assessments were made of the dendritic arbor of Golgi-impregnated pyramidal neurons in the prelimbic and anterior cingulate cortices in adulthood. RESULTS: Despite an 80% reduction in BDNF mRNA levels in both knockouts, neither spine density nor other dendritic or somal measures were decreased compared with wild-type animals. CONCLUSIONS: A reduction in BDNF expression alone does not seem to be sufficient to alter pyramidal neuron morphology in mice. This finding suggests that other molecular abnormalities are also required to produce the pyramidal neuron dendritic spine abnormalities observed in schizophrenia. PMID- 15820716 TI - Reduced hippocampal volume in unmedicated, remitted patients with major depression versus control subjects. AB - BACKGROUND: Hippocampal volumes obtained from a group of medication-free, remitted subjects with recurrent major depressive disorder (MDD) were compared against corresponding measures from healthy controls. METHODS: Thirty-one subjects with recurrent MDD in full remission, and 57 healthy controls underwent high resolution magnetic resonance imaging (MRI) on a GE 3T scanner. Eight patients with MDD were medication-naive, and twenty-three MDD patients were off antidepressant medications for a mean of 30 months at the time of the MRI study. RESULTS: Patients showed smaller total and posterior hippocampal volume relative to controls. Anterior hippocampal volume did not differ between patients and controls. CONCLUSIONS: Recurrent depression is associated with smaller hippocampal volume which is most prominent in the posterior hippocampus. Smaller hippocampal volume appears to be a trait characteristic for MDD. PMID- 15820717 TI - Estrogen receptor-beta regulates tryptophan hydroxylase-1 expression in the murine midbrain raphe. AB - BACKGROUND: Distinct expression patterns of estrogen receptor (ER)-alpha and ER beta are displayed in the murine central nervous system. ER-beta is the predominant form of the receptor expressed in the murine midbrain dorsal raphe nucleus (DRN). Tryptophan hydroxylase (TPH) is abundantly expressed in the serotonergic neurons of the DRN and is regulated by estrogen in both the monkey and the guinea pig. METHODS: In this study we used immunocytochemistry to show that ER-beta and TPH are colocalized in the serotonergic cells of the murine DRN. We utilized the ER-alpha and ER-beta gene deletion mouse models and in situ hybridization to demonstrate that ER-beta is responsible for regulating TPH1 mRNA expression. RESULTS: Estrogen increased TPH1 mRNA expression in the DRN of wild type and ER-alpha knockout mice (alpha-ERKO) but not ER-beta knockouts (beta ERKO). CONCLUSIONS: These data indicate that ER-beta is responsible for mediating estrogen regulated TPH1 expression in the murine DRN. PMID- 15820718 TI - Differential hormonal regulation of tryptophan hydroxylase-2 mRNA in the murine dorsal raphe nucleus. AB - BACKGROUND: Recently a novel tryptophan hydroxylase isoform (TPH2) was identified and shown to be highly expressed in the central nervous system (CNS). Hormonal effects on TPH2 mRNA expression in the rodent dorsal raphe nucleus (DRN) are unknown. METHODS: In situ hybridization histochemistry and real-time reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the effects of dexamethasone or estradiol on TPH2 mRNA levels in the DRN of C57/Bl6 mice. RESULTS: Dexamethasone reduced TPH2 mRNA levels in the DRN of both ovx female and intact male mice. Reduction of TPH2 mRNA in the DRN was blocked by co administration of mifepristone. Estradiol had no detectable effect on TPH2 mRNA levels in the DRN. CONCLUSIONS: TPH2 mRNA is regulated by glucocorticoids but not estradiol in the mouse DRN. Glucocorticoid-mediated reduction of TPH2 message may have relevance to the etiology of major depression, psychotic major depression in particular, where elevated glucocorticoids are one hallmark of the disease. PMID- 15820719 TI - Unreality symptoms and volumetric measures of Heschl's gyrus and planum temporal in first-episode psychosis. AB - BACKGROUND: Schizophrenia may involve dysfunction to primary auditory, speech, and language processes governed by the superior temporal gyrus (STG). These processes are implicated in hallucinations, delusions, and thought disorder. The current study explored the relationship between unreality symptoms (hallucinations and delusions) and specific STG substructures, including Heschl's gyrus (HG) and planum temporale (PT). METHODS: Twenty-five right-handed men within their first episode of psychosis were assessed using the Positive and Negative Syndrome Scale (PANSS) for the presence of hallucinations and delusional behavior (a composite score of delusions, grandiosity, suspiciousness, and unusual thought content). T1-weighted magnetic resonance imaging (MRI) scans were acquired using a 1.5 Tesla scanner. Stereological measurements of HG and PT volume were obtained. Linear regression methods explored the relationship between regional volumes and symptoms. RESULTS: Reductions in left HG were associated with hallucinations and delusions. Increases in left PT were associated with delusional behavior. CONCLUSIONS: Current results implicate HG dysfunction in unreality symptoms in men with recent-onset schizophrenia. PMID- 15820720 TI - Exposure to inorganic arsenic in drinking water and total urinary arsenic concentration in a Chilean population. AB - The relationship of inorganic arsenic exposure through drinking water and total urinary arsenic excretion in a nonoccupationally exposed population was evaluated in a cross-sectional study in three mayor cities of Chile (Antofagasta, Santiago, and Temuco). A total of 756 individuals in three population strata (elderly, students, and workers) provided first morning void urine specimens the day after exposure and food surveys were administered. Arsenic intake from drinking water was estimated from analysis of tap water samples, plus 24-h dietary recall and food frequency questionnaires. Multilevel analysis was used to evaluate the effects of the age group and city factors adjusted by predictor variables. Arsenic levels in drinking water and urine were significantly higher in Antofagasta compared with the other cities. City-and individual-level factors, 12% and 88%, respectively, accounted for the variability in urinary arsenic concentration. The main predictors of urinary arsenic concentration were total arsenic consumption through water and age. These findings indicate that arsenic concentration in drinking water continues to be the principal contributing factor to exposure to inorganic arsenic in the Chilean population. PMID- 15820721 TI - Measurements of environmental terrestrial gamma radiation dose rate in three mountainous locations in the western region of Saudi Arabia. AB - This paper describes measurements of external gamma radiation dose rate from terrestrial gamma-rays 1m above the ground in three different mountainous locations in the western region of the Kingdom of Saudi Arabia. These locations are At-Taif city, Al-Hada village, and Ash-Shafa village. CaSO4:Dy (TLD-900) thermoluminescent dosimeters were used for the detection of terrestrial gamma radiation at 40 different places in the three locations. The values of terrestrial gamma radiation dose rate measured ranged between 14 and 279 nGy h( 1) for the time interval from June 2001 to June 2002. The measured dose rate varied with the season of the year. The average gamma radiation dose rates were 468, 541, and 781 microGy y(-1) for At-Taif city, Al-Hada village, and Ash-Shafa village, respectively. The corresponding average absorbed doses to the population of the three locations were 328, 379, and 547 microSv y(-1), respectively. The quality factor of 0.7 SvG y(-1) was applied in the calculations of the absorbed dose to humans. PMID- 15820722 TI - Indoor exposures to air pollutants and allergens in the homes of asthmatic children in inner-city Baltimore. AB - This paper presents indoor air pollutant concentrations and allergen levels collected from the homes of 100 Baltimore city asthmatic children participating in an asthma intervention trial. Particulate matter (PM), NO2, and O3 samples were collected over 72 h in the child's sleeping room. Time-resolved PM was also assessed using a portable direct-reading nephelometer. Dust allergen samples were collected from the child's bedroom, the family room, and the kitchen. The mean PM10 concentration, 56.5+/-40.7 microg/m3, is 25% higher than the PM2.5 concentration (N=90), 45.1+/-37.5 microg/m3. PM concentrations measured using a nephelometer are consistent and highly correlated with gravimetric estimates. Smoking households' average PM2.5 and PM10 concentrations are 33-54 microg/m3 greater than those of nonsmoking houses, with each cigarette smoked adding 1.0 microm/m3 to indoor PM2.5 and PM10 concentrations. Large percentages of NO2 and O3 samples, 25% and 75%, respectively, were below the limit of detection. The mean NO2 indoor concentration is 31.6+/-40.2 ppb, while the mean indoor O3 concentration in the ozone season was 3.3+/-7.7 ppb. The levels of allergens are similar to those found in other inner cities. Results presented in this paper indicate that asthmatic children in Baltimore are exposed to elevated allergens and indoor air pollutants. Understanding this combined insult may help to explain the differential asthma burden between inner-city and non-inner-city children. PMID- 15820723 TI - DDT-induced oxidative damage in human blood mononuclear cells. AB - Recent work indicates that DDT and its metabolites induce apoptosis in different cellular types. However, the mechanism by which DDT generates apoptosis has not been elucidated. In this study, our data demonstrate that the apoptosis induction by DDT and its metabolites in human peripheral blood mononuclear cells (PBMC) is preceded by an increase in the levels of reactive oxygen species (ROS). Cells isolated from healthy individuals were incubated for different intervals of time (0-24 h) and in the presence of increasing concentrations of p'p-DDT, p'p-DDE, or p'p-DDD (0-80 microg/ml). The induction of oxidative stress was then determined by flow cytometry, using the compound 2',7'-dichlorofluorescein diacetate. The control level of ROS was 4.46+/-0.96 IFM, for DDT- and DDD-treated cells we obtained a 19.0-fold increment, whereas for DDE, the increment was 25-fold. ROS induction by DDT and DDE was observed after 1 h of incubation, while for DDD such levels began to be detected at 3 h of incubation; a maximum effect on the ROS production for the three compounds was found at 6 h of treatment. A significant level of ROS was induced by DDT, DDE, and DDD only at 60 and 80 microg/ml. Finally, to find an association between generation of ROS and apoptosis induction, cells incubated with DDT, DDE, and DDD were evaluated for apoptosis induction and generation of oxidative stress. Our results show that an increase in ROS was accompanied by apoptosis of PBMC in vitro. Moreover, N-acetyl-L cysteine significantly inhibits the apoptosis induction. PMID- 15820724 TI - Bioremediation of oil refinery sludge by landfarming in semiarid conditions: influence on soil microbial activity. AB - Bioremediation of a refinery sludge containing hydrocarbons in a semi-arid climate using landfarming techniques is described. The objective of this study was to assess the ability of this technique to reduce the total hydrocarbon content added to the soil with the refinery sludge in semiarid climate (low rain and high temperature). In addition, we have evaluated the effect of this technique on the microbial activity of the soil involved. For this, biological parameters (carbon fractions, microbial biomass carbon, basal respiration and ATP) and biochemical parameters(different enzymatic activities) were determined. The results showed that 80% of the hydrocarbons were eliminated in eleven months, half of this reduction taking place during the first three months. The labile carbon fractions, MBC, basal respiration and ATP of the soils submitted to landfarming showed higher values than the control soil during the first months of the process, although these values fell down by the end of the experimental period as the hydrocarbons were degraded by mineralisation. All the enzymatic activities studied: oxidoreductases such as dehydrogenase activity, and hydrolases of C(beta-glucosidase activity) and N Cycle (urease and protease) showed higher values in the soils amended with the refinery sludge than in the control. As in the case of the previous parameters, these value fell down as the bioremediation of the hydrocarbons progressed, many of them reaching levels similar to those of the control soil after eleven months. PMID- 15820725 TI - Decreased fertility in mice exposed to environmental air pollution in the city of Sao Paulo. AB - It has largely been shown that air pollution can affect human health. Effects on human fertility have been shown mainly in males by a decrease in semen quality. Few studies have focused on the environmental effects on female fertility. The aim of the present study was to analyze the effects of air pollution in the city of Sao Paulo on mouse female fertility. Four groups of female Balb/c mice were placed in two chambers 10 days (newborn) or 10 weeks (adults) after birth. Mice were maintained in the chambers 24 h a day, 7 days a week, for 4 months. The first chamber received air that had passed through an air filter (clean chamber) and the second received ambient air (polluted chamber). We measured PM10 and NO2 inside both chambers. Mice belonging to the adult groups were bred to male mice after living for 3 months inside the chambers. The newborn groups mated after reaching reproductive age (12 weeks). After 19 days of pregnancy the numbers of live-born pups, reabsorptions, fetal deaths, corpora lutea, and implantation failures were determined. PM10 and NO2 concentrations in the clean chamber were 50% and 77.5% lower than in the polluted chamber, respectively. Differences in fertility parameters between groups were observed only in animals exposed to air pollution at an early age (10 days after birth). We observed a higher number of live-born pups per animal in the clean chamber than per animal from the polluted chamber (median=6.0 and 4.0, respectively; P=0.037). There was a higher incidence of implantation failures in the polluted group than in the clean group (median=3.5 and 2.0, respectively; P=0.048). There were no significant differences in the other reproductive parameters between groups. These results support the concept that female reproductive health represents a target of air pollutants. PMID- 15820726 TI - Uptake and bioaccumulation of platinum group metals (Pd, Pt, Rh) from automobile catalytic converter materials by the zebra mussel (Dreissena polymorpha). AB - The uptake and bioaccumulation of the platinum group metals (PGM) platinum (Pt), palladium (Pd), and rhodium (Rh) by the zebra mussel (Dreissena polymorpha) were investigated in exposure studies using ground material from unused automobile catalytic converters as metal source. The mussels were exposed to the metals in tap water or humic water. In the soft tissue samples of exposed mussels mean Pt levels ranged in dependence on the type of tank water and the exposure period (6, 9, or 18 weeks) between 780 and 4300 ng/g, the Pd levels ranged between 720 and 6300 ng/g, and the Rh levels ranged between 270 and 1900 ng/g. In contrast, the control mussels had metal concentrations of <20 ng/g (Pt), <50 ng/g (Pd), and <40 ng/g (Rh). Considerably higher PGM levels were found in the exposed mussels of the humic water group than in those of the tap water group. Although there is a cumulative increase of the PGM concentrations in the environment since the introduction of the automobile catalyst more than 20 years ago, only little information about the PGM contamination in the biosphere, especially the fauna, is available. Due to the high capacity of D. polymorpha to accumulate PGM, this bivalve could be used as a potential sentinel for monitoring the noble metals in aquatic ecosystems. PMID- 15820727 TI - Lysosomal responses in the digestive gland of the freshwater mussel, Dreissena polymorpha, experimentally exposed to cadmium. AB - In order to examine the possible use of lysosomal response as a biomarker of freshwater quality, structural changes of lysosomes were measured by image analysis in the digestive gland of the zebra mussel, Dreissena polymorpha, exposed in laboratory conditions to cadmium. Mussels were exposed to the metal (10 and 200 microg/L) for 3 weeks and randomly collected after 7 and 21 days. At each treatment day, digestive tissues were excised and beta-glucuronidase activity was revealed in cryotome sections. Four stereological parameters were calculated: lysosomal volume density, lysosomal surface density, lysosomal surface to volume ratio, and lysosomal numerical density. The changes observed in this study reflected a general activation of the lysosomal system, including an increase in both the number and the size of lysosomes in the digestive gland cells of mussels exposed to cadmium. The digestive lysosomal response in zebra mussels was related to exposure time and to metal concentration, demonstrating the potential of this biomarker in freshwater biomonitoring. PMID- 15820728 TI - Iron status influences trace element levels in human blood and serum. AB - Food is the main source of trace elements for the general population. The gastrointestinal absorption of certain trace elements, e.g., cadmium, is strongly influenced by iron (Fe) status. This factor may also be relevant for the bioavailability of other trace elements. Therefore, we investigated relationships between Fe status indicators and trace element concentrations in blood and serum of 234 boys and girls at ages 15 and 17 years. Fe status was measured using serum ferritin (S-Ft), soluble transferrin receptor in serum (sTfR), and the ratio sTfR/S-Ft. The trace elements we investigated were, in blood, cadmium, cobalt, copper, zinc, selenium, rubidium, mercury, and lead, and, in serum, cobalt, copper, zinc, selenium, rubidium, tungsten, mercury, and lead. We found inverse correlations between Fe status and blood cadmium, blood or serum cobalt, or blood copper. There were positive correlations between Fe status and mercury concentrations. Selenium was positively correlated with sTfR. The relationships between Fe status and lead were equivocal. There were fewer correlations for serum than for blood, but the inverse relationships between Fe status and cobalt were equally strong in serum and blood. We found only occasional, and perhaps spurious, correlations with zinc, rubidium, and tungsten. In conclusion, previous indications that cadmium, cobalt, and copper are absorbed by transport mechanisms similar to that of Fe are supported by this study. Strong positive correlations between Fe status and mercury concentrations remain to be explained. PMID- 15820729 TI - Cancer incidence and asbestos in drinking water, Town of Woodstock, New York, 1980-1998. AB - Late in 1985, asbestos contamination was discovered in the public water supply of the Town of Woodstock, Ulster County, New York. Contamination resulted from asbestos-cement pipes installed in the town water system in the mid to late 1950s and the corrosiveness of the local water. The New York State (NYS) Department of Health established the Woodstock Asbestos Exposure Registry (WAER) in 1986 to monitor rates of cancer among individuals who lived on the water supply between 1960 and 1985. Demographic, health, and residential information were collected on 2936 registrants. The follow-up period for observation of cancer was 1980-1998, consistent with the expected lag of 20-30+ years for development of asbestos related cancers. The NYS Cancer Registry was used to ascertain cancer diagnoses. Standardized incidence ratios (SIRs) for gastrointestinal, respiratory, and total cancers were all approximately 1.00 or less and all 95% confidence intervals (CIs) included 1.00. For individual types of the gastrointestinal cancers, only the SIR for pancreatic cancer was marginally statistically significant at 2.19 (95% CI=1.00-4.16), based on a total of nine observed cases. The excess in pancreatic cancer occurred primarily among men (SIR=3.08; 95% CI=1.13-6.70) and was only slightly elevated among women (SIR=1.39; 95% CI=0.29-4.06). This association may be related to factors other than asbestos exposure such as occupation and lifestyle or to chance. No cases of mesothelioma were observed among WAER participants. There was no increase in incidence by latency or duration of residence on the water supply, but the ability to detect these trends is limited by small numbers and unknown dates of initial exposure. The general pattern of results did not demonstrate a likely link between exposure to asbestos in drinking water and cancer occurrence among participants in the WAER. PMID- 15820730 TI - Endosulfan and its metabolites in fertile women, placenta, cord blood, and human milk. AB - Although industrialized nations have restricted or banned many organochlorine pesticides, some of these chemicals (e.g., endosulfans) are still used, on the assumption that they pose little threat to the environment, wildlife, or human health. According to available information, Spain is the main consumer of endosulfans within the European Union, accounting for almost half of the total consumption. Reports on human exposure in Southern Spain to persistent bioacumulable organochlorine pesticides have indicated considerable exposure to endosulfans. The present study investigated the presence of endosulfan I, endosulfan II, and endosulfan metabolites in fatty and non-fatty tissues and fluids from women of reproductive age and children in Southern Spain. The highest concentration of commercial endosulfan I and endosulfan II was found in adipose tissue, with a mean value (I+II) of 17.72 ng/g lipid, followed by human milk, with a mean value (I+II) of 11.38 ng/mL milk. These findings support the lipophilicity of these chemicals and their elimination by milk secretion. The concentration in the placenta homogenate was similar to that in the blood from the umbilical cord (7.74 and 6.11 ng/mL, respectively) and reflected their lower fat content. Endosulfan diol and endosulfan sulfate were more frequently found in placenta homogenate, with a mean concentration of 12.56 and 3.57 ng/mL, respectively, and in blood from umbilical cord, at 13.23 and 2.82 ng/mL, respectively. Therefore, women of reproductive age in Southern Spain appear to be currently exposed to endosulfans. Because these chemicals can be mobilized during pregnancy and lactation, further research is warranted to investigate the health consequence in children resulting from exposure to chemicals suspected of immunotoxic, neurotoxic, or endocrine-disrupting effects. PMID- 15820731 TI - Traffic at residential address, respiratory health, and atopy in adults: the National German Health Survey 1998. AB - Motor vehicle traffic contributes to more than 50% of PM10 in Europe and might have far reaching impacts on human health. We investigated the relationship between residential street type as a surrogate for traffic intensity and the prevalence of respiratory symptoms, atopic diseases, and allergic sensitization in adults. Data from 6896 subjects of the German Health Survey 1998 with complete information on residential street type were used. Multiple logistic regression analyses were applied to model associations between street type categories, and respiratory and atopic outcomes were assessed by screening questionnaire of The European Respiratory Health Survey and specific IgE measurements. Living at extremely or considerably busy roads (23.9% of total study population) compared to roads with no or rare traffic (64.5%) was statistically significantly associated with chronic bronchitis (aOR 1.36 (95% CI) (1.01-1.83)) while nocturnal coughing attacks (past 12 months) (1.24 (0.98-1.57)), wheeze during the past 12 months (1.21 (0.93-57)), and hay fever (1.16 (0.94-1.42)) were marginally increased after adjustment for several potential confounders and for multiple testing. No increased risks were found for asthma (0.97 (0.67-1.42)) and allergic sensitization (1.05 (0.91-1.20)). We conclude that exposure to traffic-related air pollutants increases the risk of nonallergic respiratory symptoms and to a lesser degree the risk of hay fever and allergic sensitization but not the risk of asthma in adults. PMID- 15820732 TI - Biomonitoring of organochlorines in women with benign and malignant breast disease. AB - Established risk factors for breast cancer explain breast cancer risk only partially. Organochlorines are considered to be a possible cause for hormone dependent cancers. A hospital-based case-control study, the first from India, was conducted among 50 women undergoing surgery for breast disease to examine the association between organochlorine exposure and breast cancer risk. Blood, tumor, and surrounding adipose tissue of the breast were collected from the subjects with benign (control) and malignant breast (study) lesions and analyzed to determine organochlorine insecticides using a gas-liquid chromatograph equipped with an electron capture detector. The alpha, beta, gamma, and delta isomers of hexachlorocyclohexane (HCH), p,p'-dichlorodiphenyltrichloroethane (DDT), o,p' DDT, p,p-dichlorodiphenyldichloroethylene, and p,p' dichlorodiphenyldichloroethane were frequently detected in three specimens. Total HCH and total DDT levels were higher in the blood of the study group (25 cases) than in those of the controls (25 cases) with only gamma-HCH being significantly different (P<0.05). However, both total HCH and total DDT were higher in the tumor tissues of the controls than in those of the study group; gamma-HCH was significantly different (P<0.05). The level of total HCH (alpha-HCH was significantly different, P<0.05) was higher in the breast adipose tissue of the study group, whereas total DDT was higher in the breast adipose tissue of the control group. The distribution of known confounders of breast cancer including age, body mass index, age at menarche and menopause, duration of breast feeding, and family history related to breast disease did not differ significantly between benign and malignant groups. This pilot study with limited statistical power does not support a positive association between exposure to organochlorines and risk of breast cancer but paves the way for a larger Indian study with greater statistical power encompassing different regions of the country to enable statistically sound conclusions. PMID- 15820733 TI - An examination of the trade-offs in public health resulting from the use of default exposure assumptions in fish consumption advisories. AB - Efforts to provide for public health protection from environmental contaminants in fish have resulted in various advisories or recommendations with regard to fish consumption from local, state, and federal agencies. These advisories are based on measured levels of contaminants in fish that are combined with values for body weight and portion size to produce an estimate of an "acceptable" consumption frequency (e.g., eat no more than once per month). Because values for body weight and portion size are generally generic default values, they do not necessarily relate to a specific population or to any individual in that population. Thus, the use of default values may result in underprotection or overprotection in any given case. Given the benefits of fish consumption and the risks from overexposure to various toxicants, vigilance is required by custodians of public health to ensure that populations are protected while being cautious not to over- or underprotect them. In this analysis, we examine the "acceptable" consumption limits derived for fish species/groups consumed by three specific populations and determine the extent of public health protection afforded by these limits. To accomplish this, the "acceptable" consumption frequencies are derived based, in part, on default assumptions and are compared to intakes calculated from empirically derived species-specific individual consumption and demographic data. Sensitivity analyses and population-specific probabilistic assessments of exposure are conducted to identify those values and/or assumptions which might significantly influence the resulting fish consumption advisories. Three populations were chosen for study based on their ability to represent populations of greatest concern: those most sensitive and/or those most exposed. We conclude from this investigation that consumption pattern data, contaminant data and body weight data together can be used to make fish consumption advisories more focused and, therefore, less likely to be under- or overprotective. PMID- 15820734 TI - Fishing, fish consumption, and knowledge about advisories in college students and others in central New Jersey. AB - Risks to humans and other organisms from consuming fish have become a national concern. Over the past 3 years there have been a number of national advisories regarding saltwater fish. Although information on fishing, consumption patterns, and public knowledge about advisories has been examined for at-risk populations, there is little information about the latter for a general population. Overall knowledge about advisories, ratings for information about the risks and benefits of eating fish, and the relationship between fishing, consumption patterns, and knowledge about advisories was examined in a sample of 180 college students and others residing in central New Jersey, USA. The null hypothesis of no differences in fishing, consumption, and knowledge about advisories as a function of age, gender, ethnicity, and education was tested. A significantly higher proportion of men fished compared to women, and significantly fewer Blacks and Asians fished than did Whites or Hispanics. More Asians who fished did so in salt water, compared to others. There were no gender differences in consumption patterns, and few age or ethnic differences, mainly due to low sample sizes in some ethnic groups. Significantly fewer young people and fewer Asians ate fish compared to others. Overall, more people knew about the benefits of eating fish than the risks. Half as many people had heard about advisories concerning tuna, and less than a third knew about advisories concerning shark and swordfish than had heard general warnings. There were no gender differences in knowing about advisories, but there were several ethnic differences: a lower percentage of Asians generally knew that there were advisories, and fewer Blacks knew that there were benefits from eating fish than others. People in the age group 21-45 years were less aware of both the benefits and the risks from eating fish compared to older people. These data suggest that risk managers need to target younger people for information about the risks and benefits of consuming fish, particularly given that this is the population that will become pregnant over the next few years. Contrary to some previous research, subjects trusted family, friends, and other fishermen less for knowledge about the risks and benefits of fish consumption than other sources, such as doctors, governmental officials, and university professors. Even with this limited sample size, it is clear that people have heard more about the benefits of fish consumption than the risks, and a relatively low percentage have heard about the recent US Food and Drug Administration warnings about swordfish and shark. PMID- 15820735 TI - Science-based risk assessments for drinking water disinfection by-products. PMID- 15820736 TI - A stochastic automata network descriptor for Markov chain models of instantaneously coupled intracellular Ca2+ channels. AB - Although there is consensus that localized Ca(2+) elevations known as Ca(2+) puffs and sparks arise from the cooperative activity of intracellular Ca(2+) channels, the precise relationship between single-channel kinetics and the collective phenomena of stochastic Ca(2+) excitability is not well understood. Here we present a formalism by which mathematical models for Ca(2+)-regulated Ca(2+) release sites are derived from stochastic models of single-channel gating that include Ca(2+) activation, Ca(2+) inactivation, or both. Such models are stochastic automata networks (SANs) that involve a large number of functional transitions, that is, the transition probabilities of the infinitesimal generator matrix of one of the automata (i.e., an individual channel) may depend on the local [Ca(2+)] and thus the state of the other channels. Simulation and analysis of the SAN descriptors representing homogeneous clusters of intracellular Ca(2+) channels show that (1) release site density can modify both the steady-state open probability and stochastic excitability of Ca(2+) release sites, (2) Ca(2+) inactivation is not a requirement for Ca(2+) puffs or sparks, and (3) a single channel model with a bell-shaped open probability curve does not lead to release site activity that is a biphasic function of release site density. These findings are obtained using iterative, memory-efficient methods (novel in this biophysical context and distinct from Monte Carlo simulation) that leverage the highly structured SAN descriptor to unambiguously calculate the steady-state probability of each release site configuration and puff statistics such as puff duration and inter-puff interval. The validity of a mean field approximation that neglects the spatial organization of Ca(2+) release sites is also discussed. PMID- 15820737 TI - Interaction of the IP3-Ca2+ and MAPK signaling systems in the Xenopus blastomere: a possible frequency encoding mechanism for the control of the Xbra gene expression. AB - The intense periodic calcium activity experimentally observed in the Xenopus embryo at the Mid Blastula Transition stage is closely related to the competence of the embryonic cells of the marginal zone to respond to the posterior mesodermal inducting signals from the Fibroblast Growth Factor (FGF). In this work we do a stability analysis and study numerically an extension of a mathematical model previously introduced by us [Diaz, J., Baier, G., Martinez Mekler, G., Pastor, N., 2002. Interaction of the IP(3)-Ca(2+) and the FGF-MAPK signaling pathways in the Xenopus laevis embryo: a qualitative approach to the mesodermal induction problem. Biophys. Chem. 97, 55-72] for the interaction of the Inositol 1,4,5-triphosphate-Calcium (IP(3)-Ca(2+)) and the Mitogen-Activated Protein Kinase (MAPK) signaling pathways at the Mid Blastula Transition stage or stage 8 of development. This allows us to consider the effect of the oscillatory calcium dynamics on the FGF input signal carried by the MAP kinase (ERK) into the nucleus. We find that this interaction of the pathways induces a limit cycle behavior for ERK with frequency-encoding characteristics. We believe that this periodic increase of the ERK levels in the nucleus is related to the ability of the cell to express posteriorizing mesodermal features induced by the FGF signal at stage 8. PMID- 15820738 TI - The role of seasonality in the dynamics of deer tick populations. AB - In this paper, we formulate a nonlinear system of difference equations that models the three-stage life cycle of the deer tick over four seasons. We study the effect of seasonality on the stability and oscillatory behavior of the tick population by comparing analytically the seasonal model with a non-seasonal one. The analysis of the models reveals the existence of two equilibrium points. We discuss the necessary and sufficient conditions for local asymptotic stability of the equilibria and analyze the boundedness and oscillatory behavior of the solutions. A main result of the mathematical analysis is that seasonality in the life cycle of the deer tick can have a positive effect, in the sense that it increases the stability of the system. It is also shown that for some combination of parameters within the stability region, perturbations will result in a return to the equilibrium through transient oscillations. The models are used to explore the biological consequences of parameter variations reflecting expected environmental changes. PMID- 15820739 TI - Dynamics of the 'echo' effect in a phytoplankton system with nitrogen fixation. AB - Consideration of nitrogen fixation adds a positive nonlinear feedback to plankton ecosystem models. We investigate the consequences of this feedback for secondary phytoplankton blooms and the response of phytoplankton dynamics to physical forcing. The dynamics of phytoplankton, Trichodesmium (the nitrogen fixer), and nutrients is modeled with a system of three differential equations. The model includes two types of nonlinear interactions: the competition of phytoplankton and Trichodesmium for light, and the positive feedback resulting from Trichodesmium recycling. A typical simulation of the model in time, with forcing by a varying mixed-layer depth, reveals a clear successional sequence including a secondary or 'echo' bloom of the phytoplankton. We explain this sequence of events through the stability analysis of three different steady states of the model. Our analysis shows the existence of a critical biological parameter, the ratio of normalized growth rates, determining the occurrence of 'echo' blooms and the specific sequence of events following a physical perturbation. The interplay of positive and negative feedbacks appears essential to the timing and the type of events following such a perturbation. PMID- 15820740 TI - Mathematical models for the Aedes aegypti dispersal dynamics: travelling waves by wing and wind. AB - Biological invasion is an important area of research in mathematical biology and more so if it concerns species which are vectors for diseases threatening the public health of large populations. That is certainly the case for Aedes aegypti and the dengue epidemics in South America. Without the prospect of an effective and cheap vaccine in the near future, any feasible public policy for controlling the dengue epidemics in tropical climates must necessarily include appropriate strategies for minimizing the mosquito population factor. The present paper discusses some mathematical models designed to describe A. aegypti's vital and dispersal dynamics, aiming to highlight practical procedures for the minimization of its impact as a dengue vector. A continuous model including diffusion and advection shows the existence of a stable travelling wave in many situations and a numerical study relates the wavefront speed to a few crucial parameters. Strategies for invasion containment and its prediction based on measurable parameters are analysed. PMID- 15820741 TI - Probabilistic Gompertz model of irreversible growth. AB - Characterizing organism growth within populations requires the application of well-studied individual size-at-age models, such as the deterministic Gompertz model, to populations of individuals whose characteristics, corresponding to model parameters, may be highly variable. A natural approach is to assign probability distributions to one or more model parameters. In some contexts, size at-age data may be absent due to difficulties in ageing individuals, but size increment data may instead be available (e.g., from tag-recapture experiments). A preliminary transformation to a size-increment model is then required. Gompertz models developed along the above lines have recently been applied to strongly heterogeneous abalone tag-recapture data. Although useful in modelling the early growth stages, these models yield size-increment distributions that allow negative growth, which is inappropriate in the case of mollusc shells and other accumulated biological structures (e.g., vertebrae) where growth is irreversible. Here we develop probabilistic Gompertz models where this difficulty is resolved by conditioning parameter distributions on size, allowing application to irreversible growth data. In the case of abalone growth, introduction of a growth limiting biological length scale is then shown to yield realistic length increment distributions. PMID- 15820742 TI - Stochastic vs. deterministic uptake of dodecanedioic acid by isolated rat livers. AB - Deterministic and stochastic differential equations models of the uptake of dodecanedioic acid (C12) are fitted to experimental data obtained on nine isolated, perfused rat livers. 11500 mug of C12 were injected as a bolus into the perfusing liver solution. The concentrations of C12 in perfusate samples taken over 2 h from the beginning of the experiments were analyzed by High Performance Liquid Chromatography (HPLC). A two-compartment deterministic model is studied. To include spontaneous erratic variations in the metabolic processes the parameter for the uptake rate is randomized to obtain a stochastic differential equations model. Parameters are estimated in a two-step procedure: first, parameters in the drift part are estimated by least squares; then, the diffusion parameter is estimated using Monte-Carlo simulations to approximate the unknown likelihood function. Parameter estimation is carried out over a wide range of reasonable measurement error variances to check robustness of estimates. It is concluded that the kinetics of dodecanedioic acid, in the experimental conditions discussed, is well approximated by a model including spontaneous erratic variations in the liver uptake rate. PMID- 15820743 TI - The mathematical modelling of adjuvant chemotherapy scheduling: incorporating the effects of protocol rest phases and pharmacokinetics. AB - In this paper the modelling objective is to determine the drug alternation time which minimises the formation of resistant tumour cells when delivering two non cross resistant chemotherapeutics given such drugs cannot be delivered simultaneously and constraints due to pharmacokinetics and protocol rest phases. We initially consider cell cycle phase non-specific models, as investigated by Goldie and Coldman. By extending previous work, these models are generalised to consider chemotherapeutic S-phase specificity. We find with the cell cycle phase non-specific models that once the alternation time of the drugs is reduced below a critical threshold, a substantial improvement in protocol outcome is predicted. Extensive improvements are also observed for the S-phase specific investigation if the drugs can be alternated extremely rapidly. However, this is typically impossible due to pharmacokinetic constraints. Under such circumstances, the most appropriate choice of the alternation time can depend sensitively on the median and variance of the tumour cell cycle time in a complicated manner. For schedulings motivated by Capecitabine protocols, we find that switching the drugs only once, or at most twice, between rest phases gives the most reliable alternation time. The main and novel conclusion of this paper is the modelling prediction that one must be much more specific in the choice of the protocol alternation time if attempting to observe the improvements promised by Goldie and Coldman's alternation hypothesis for the rest phases, pharmacokinetics and delivery mechanisms typically encountered in cell cycle phase specific chemotherapy protocols. PMID- 15820744 TI - Thermodynamical interpretation of evolutionary dynamics on a fitness landscape in an evolution reactor, II. AB - In our previous report [Aita, T., Morinaga, S., Hosimi, Y., 2004. Thermodynamical interpretation of evolutionary dynamics on a fitness landscape in an evolution reactor I. Bull. Math. Biol. 66, 1371-1403], an analogy between thermodynamics and adaptive walks on a Mt. Fuji-type fitness landscape in an artificial selection system was presented. Introducing the 'free fitness' as the sum of a fitness term and an entropy term and 'evolutionary force' as the gradient of free fitness on a fitness coordinate, we demonstrated that the adaptive walk (=evolution) is driven by the evolutionary force in the direction in which free fitness increases. In this report, we examine the effect of various modifications of the original model on the properties of the adaptive walk. The modifications were as follows: first, mutation distance d was distributed obeying binomial distribution; second, the selection process obeyed the natural selection protocol; third, ruggedness was introduced to the landscape according to the NK model; fourth, a noise was included in the fitness measurement. The effect of each modification was described in the same theoretical framework as the original model by introducing 'effective' quantities such as the effective mutation distance or the effective screening size. PMID- 15820745 TI - Regimes of biological invasion in a predator-prey system with the Allee effect. AB - Spatiotemporal dynamics of a predator-prey system is considered under the assumption that prey growth is damped by the strong Allee effect. Mathematically, the model consists of two coupled diffusion-reaction equations. The initial conditions are described by functions of finite support which corresponds to invasion of exotic species. By means of extensive numerical simulations, we identify the main scenarios of the system dynamics as related to biological invasion. We construct the maps in the parameter space of the system with different domains corresponding to different invasion regimes and show that the impact of the Allee effect essentially increases the system spatiotemporal complexity. In particular, we show that, as a result of the interplay between the Allee effect and predation, successful establishment of exotic species may not necessarily lead to geographical spread and geographical spread does not always enhance regional persistence of invading species. PMID- 15820746 TI - IL-1beta stimulates argininosuccinate synthetase gene expression through NF kappaB in Caco-2 cells. AB - Argininosuccinate synthetase (ASS) is limiting the arginine synthesis and can be stimulated by immunostimulants. We previously identified a putative NF-kappaB element in the human ASS gene promoter but its functionality was unknown (Husson et al., Eur. J. Biochem. 270 (2003) 1887). In the present study, using Caco-2 cells, a human enterocyte line, we demonstrate that IL-1beta rapidly induces the expression of the ASS gene at a transcriptional level through NF-kappaB activation. Using gel shift assay and double-strand oligonucleotide sequence of the identified putative NF-kappaB binding site of the ASS promoter, we provide evidence that NF-kappaB may functionally interact with this element. PMID- 15820747 TI - Characterization of Sparus aurata osteonectin cDNA and in silico analysis of protein conserved features: evidence for more than one osteonectin in Salmonidae. AB - Osteonectin is a matricellular protein involved in various cellular mechanisms but its exact function remains unclear despite numerous studies. We present here the cloning of Sparus aurata partial osteonectin cDNA and the reconstruction of 15 other sequences from both vertebrates and invertebrates, almost doubling the set of available sequences (a total of 35 sequences is now available). Taking advantage of the resulting large amount of data, we have created multiple sequence alignments and identified osteonectin putative conserved features (intra and inter-disulfide bonds, collagen- and calcium-binding domains and phosphorylation sites) likely to be important for protein structure and function. This work also provides the first evidence for the presence of more than one osteonectin in some species. Finally, S. aurata osteonectin gene expression has been shown to initiate during larval development shortly after gastrulation, and to be high in bone-derived cell lines while down-regulated during extracellular matrix mineralization, further emphasizing the important role of osteonectin in skeletal development and bone formation. PMID- 15820748 TI - Functional expression of pig renal organic anion transporter 3 (pOAT3). AB - With the cloning of pig renal organic anion transporter 1 (pOAT1) (Biochimie 84 (2002) 1219) we set up a model system for comparative studies of cloned and natively isolated membrane located transport proteins. Meanwhile, another transport protein involved in p-aminohippurate (PAH) uptake on the basolateral side of the proximal tubule cells was identified, designated organic anion transporter 3 (OAT3). To explore the contribution of pOAT1 to the PAH clearance in comparison to OAT3, it was the aim of this study to extend our model by cloning of the pig ortholog of OAT3. Sequence comparisons of human organic anion transporter 3 (hOAT3) with the expressed sequence tag (EST) database revealed a clone and partial sequence of the pig renal organic anion transporter 3 (pOAT3) ortholog. Sequencing of the entire open reading frame resulted in a protein of 543 amino acid residues encoded by 1632 base pairs (EMBL Acc. No. AJ587003). It showed high homologies of 81%, 80%, 76%, and 77% to the human, rabbit, rat, and mouse OAT3, respectively. A functional characterization of pOAT3 in Xenopus laevis oocytes yielded an apparent Km (Kt) for [3H]estrone sulfate of 7.8 +/- 1.3 microM. Moreover, pOAT3 mediated [3H]estrone sulfate uptake was almost abolished by 0.5 mM of glutarate, dehydroepiandosterone sulfate, or probenecid consistent with the hallmarks of OAT3 function. PMID- 15820749 TI - Amino terminal peptides from the Plasmodium falciparum EBA-181/JESEBL protein bind specifically to erythrocytes and inhibit in vitro merozoite invasion. AB - Several EBA-175 paralogues (EBA-140, EBA-165, EBA-175, EBA-181, and EBL-1) have been described among the Plasmodium falciparum malaria parasite proteins, which are important in the red blood cell (RBC) invasion process. EBA-181/JESEBL is a 181 kDa protein expressed in the late schizont stage and located in the micronemes; it belongs to the Plasmodium Duffy binding-like family and is able to interact with the erythrocyte surface. Here, we describe the synthesis of 78, 20 mer synthetic peptides derived from the reported EBA-181/JESEBL sequence and their ability to bind RBCs in receptor-ligand assays. Five peptides (numbered 30030, 30031, 30045, 30051, and 30060) displayed high specific binding to erythrocytes; their equilibrium binding parameters were then determined. These peptides interacted with 53 and 33 kDa receptor proteins on the erythrocyte surface, this binding being altered when RBCs were pretreated with enzymes. They were able to inhibit P. falciparum merozoite invasion of RBCs when tested in in vitro assays. According to these results, these five EBA-181/JESEBL high specific erythrocyte binding peptides, as well as the entire protein, were seen to be involved in the molecular machinery used by the parasite for invading RBCs. They are thus suggested as potential candidates in designing a multi-sub-unit vaccine able to combat the P. falciparum malaria parasite. PMID- 15820750 TI - Relationship between the inhibition of phosphatidic acid phosphohydrolase-1 by oleate and oleoyl-CoA ester and its apparent translocation. AB - Phosphatidic acid phosphohydrolase-1 (PAP-1) activity is reversibly inhibited by fatty acids and their acyl-CoA esters and it appears paradoxical that these effectors have been reported to increase the liver's esterification capacity by translocating the rate-limiting enzyme PAP-1 from cytosol to the endoplasmic reticulum. Therefore, we have examined the effect of oleate, oleoyl-CoA, and spermine on the activation and translocation of PAP-1 of rat liver. PAP-1 activity is directly inhibited by oleic acid and oleoyl-CoA ester in an allosteric manner, resulting in the formation of inactive PAP-1-fatty acid (or acyl-CoA) complex, even in the absence of any subcellular structures. Such association/aggregation of PAP-1 can be easily collected by centrifugation and may explain the apparent translocation phenomenon of this enzyme to a particular structure in the presence of fatty acids or acyl-CoA esters as reported in many works. Indeed, incubation of cytosol fraction alone with oleate or oleoyl-CoA at 37 degrees C, followed by centrifugation, induces a significant increase (sevenfold) in PAP-1 activity in the pellet fraction. This displacement is accompanied by an increase in the specific activity of PAP-1 in the pellet fraction. Spermine is less effective than oleate in inducing the displacement of PAP-1 activity from cytosol to the pellet fraction in the absence of any membrane structures. This apparent translocation of PAP-1 is also promoted when homogenate fraction was incubated with oleate prior to the preparation of cytosol and microsomal fraction. Thus, many of the announced factors, including fatty acids, would promote the in vitro association/aggregation of PAP-1 enzyme rather than its translocation, and therefore, re-evaluation of the reported effects on PAP-1 translocation phenomenon is required. It is proposed that fatty acids and their esters would favour beta-oxidation over esterification by promoting the forming of inactive associated PAP-1 in situations such as starvation and metabolic stress in which there is an increased supply of fatty acids to the liver. PMID- 15820751 TI - Molecular characterization of a glutathione transferase from Pinus tabulaeformis (Pinaceae). AB - Glutathione transferases (GSTs) play important roles in stress tolerance and detoxification metabolism in plants. To date, studies on GSTs in higher plants have focused largely on agricultural plants. In contrast, there is virtually no information on the molecular characteristics of GSTs in gymnosperms. The present study reports for the first time the cloning, expression and characteristics of a GST gene (PtGSTU1) from a pine, Pinus tabulaeformis, which is widely distributed from northern to central China covering cold temperate and drought regions. The PtGSTU1 gene encodes a protein of 228 amino acid residues with a calculated molecular mass of 26.37 kDa. Reverse transcription PCR revealed that PtGSTU1 was expressed in different tissues, both above and below ground, of P. tabulaeformis. The over-expressed recombinant PtGSTU1 showed high activity towards the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 7-chloro-4-nitrobenzo-2-oxa-1,3 diazole (NBD-Cl). Kinetic analysis with respect to CDNB as substrate revealed a Km of 0.47 mM and Vmax of 169.1 micromol/min per mg of protein. The recombinant PtGSTU1 retained more than 60% of its maximum enzymatic activity from 15 degrees C to 45 degrees C with a broad optimum Tm range of 25 degrees C - 35 degrees C. The enzyme had a maximum activity at approximately pH 8.5 - 9.0. Site-directed mutagenesis revealed that Ser13 in the N-terminal domain is a critical catalytic residue, responsible for stabilisation of the thiolate anion of enzyme-bound glutathione. Based on comparative analyses of its amino acid sequence, phylogeny and predicted three-dimensional structure, the PtGSTU1 should be classified as a tau class GST. PMID- 15820752 TI - Green tea modulation of inducible nitric oxide synthase in hypoxic/reoxygenated cardiomyocytes. AB - Hypoxia/reoxygenation (H/R) is one of the causes of the increased expression of inducible nitric oxide synthase (iNOS) in cardiomyocytes. Since an aberrant NOS induction has detrimental consequences, we evaluated the effect of a green tea extract (GTE) on the NOS induction and activity in H/R-cardiomyocytes to define a nutritional strategy. Cultured rat cardiomyocytes were exposed to H/R in the presence of two concentrations of a green tea extract (GTE), which is reported to inhibit NOS expression and activity in different cells. In cultured cardiomyocytes two NOS isoforms were constitutively expressed, but only iNOS was induced by H/R. GTE supplementation at the lowest concentration, comparable to that in human plasma after dietary consumption, was ineffective, while the highest, comparable to that achievable by dietary supplements, counteracted the effect of H/R on iNOS induction and activity. It is necessary to verify in humans the relationship between the modulation of NO production and green tea dietary consumption. PMID- 15820753 TI - Identifying Plasmodium falciparum merozoite surface protein-10 human erythrocyte specific binding regions. AB - Receptor-ligand interactions between synthetic peptides and normal human erythrocytes were studied to determine P. falciparum merozoite surface protein-10 (MSP-10) regions specifically binding to membrane surface receptors on human erythrocytes. Three MSP-10 protein High Activity Binding Peptides (HABPs) were identified, whose binding to erythrocytes became saturable and sensitive on being treated with neuraminidase, trypsin and chymotrypsin. Some of them specifically recognised a 50 kDa erythrocyte membrane protein. Some HABPs inhibited in vitro P. falciparum merozoite invasion of erythrocytes by 70%, suggesting that MSP-10 protein's possible role in the invasion process probably functions by using similar mechanisms to those described for other MSP family antigens. In addition to above results, the high homology in amino-acid sequence and superimposition of both MSP-10, MSP-8 and MSP-1 EGF-like domains and HABPs 31132, 26373 and 5501 suggest that tridimensional structure could be playing an important role in the invasion process and in designing synthetic multi-stage anti-malarial vaccines. PMID- 15820754 TI - Calorimetry and mass spectrometry study of oxidized calmodulin interaction with target and differential repair by methionine sulfoxide reductases. AB - Calmodulin is known to be a target for oxidation, which leads to conversion of methionine residues to methionine sulfoxides. Previously, we reported that both methionine sulfoxide reductases MsrA and MsrB were able to reduce methionine sulfoxide residues in oxidized calmodulin. In the present study, we have made use of the interaction between calmodulin and RS20, a peptide model for calmodulin targets, to probe the structural consequences of oxidation and mode of repair both by MsrA and MsrB. Isothermal titration calorimetry and differential scanning calorimetry showed that oxidized calmodulin interacts with RS20 via its C terminal domain only, resulting in a non-productive complex. As shown by spectrofluorometry, oxidized calmodulin treated with MsrA exhibited native binding affinity for RS20. In contrast, MsrB-treatment of oxidized calmodulin resulted in 10-fold reduced affinity. Mass spectrometry revealed that the sulfoxide derivative of methionine residue 124 was differentially repaired by MsrA and MsrB. This provided a basis for rationalizing the difference in binding affinities of oxidized calmodulin reported above, since Met124 residue had been shown to be critical for interaction with some targets. This study provides the first evidence that in an oxidized polypeptide chain MetSO residues might be differentially repaired by the two Msr enzymes. PMID- 15820755 TI - Tat-mediated protein transduction of human brain pyridoxal kinase into PC12 cells. AB - Pyridoxal kinase (PK) catalyses the phosphorylation of vitamin B6 to pyridoxal-5' phosphate (PLP). A human brain PK gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-PK fusion protein. The expressed and purified Tat-PK fusion proteins transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced Tat-PK proteins showed catalytic activity and are stable for 48 h. The intracellular concentration of PLP, which is known as a biologically active form of vitamin B6, was increased by pre-treatment of Tat-PK to the PC12 cells. Those results suggest that the transduction of Tat-PK fusion protein can be one of the ways to regulate the PLP level and to replenish this enzyme in the various neurological disorders related to vitamin B6. PMID- 15820756 TI - Carbon-11 HOMADAM: a novel PET radiotracer for imaging serotonin transporters. AB - Carbon-11-labeled N,N-dimethyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine (HOMADAM) was synthesized as a new serotonin transporter (SERT) imaging agent. METHODS: Carbon-11 was introduced into HOMADAM by preparation of N-methyl-2-(2' amino-4'-hydroxymethylphenylthio)benzylamine followed by alkylation with carbon 11 iodomethane. Binding affinities of HOMADAM and the radiolabeling substrate, N methyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine, were determined in cDNA transfected cells expressing human SERT, dopamine transporters (DAT) and norepinephrine transporters NET using [3H]citalopram, [(125)I]RTI-55 and [3H]nisoxetine, respectively. MicroPET brain imaging was performed in monkeys. Arterial plasma metabolites of HOMADAM were analyzed in a rhesus monkey by high performance liquid chromatography (HPLC). RESULTS: HOMADAM displayed high affinity for the SERT (Ki = 0.6 nM). N-methyl-2-(2'-amino-4' hydroxymethylphenylthio)benzylamine displayed moderate affinity for the SERT (Ki = 15.11 nM). The affinities of HOMADAM for the DAT and NET were 2000- and 253 fold lower, respectively, than for the SERT. [11C]HOMADAM was prepared from [11C]iodomethane in approximately 25% radiochemical yield (decay-corrected to end of bombardment). MicroPET brain imaging studies in monkeys demonstrated that [11C]HOMADAM uptake was selectively localized in the midbrain, thalamus, pons, caudate, putamen and medulla. The midbrain-to-cerebellum, pons-to-cerebellum, thalamus-to-cerebellum and putamen-to-cerebellum ratios at 85 min were 4.2, 2.8, 2.3 and 2.0, respectively. HOMADAM binding achieved quasi-equilibrium at 45 min. Radioactivity in the SERT-rich regions of monkey brain was displaceable with R,S citalopram. Radioactivity in the DAT-rich regions of monkey brain was not displaceable with the DAT ligand RTI-113. Radioactivity in the SERT-rich regions of monkey brain was displaceable with the R,S-reboxetine, a NET ligand with a high nanomolar affinity for SERT. Arterial plasma metabolites of HOMADAM were analyzed in a rhesus monkey by HPLC and displayed a single peak that corresponded to unmetabolized HOMADAM. CONCLUSION: HOMADAM is an excellent candidate for PET primate imaging of brain SERTs. PMID- 15820757 TI - Synthesis and in vitro and in vivo evaluation of three radioiodinated nitroimidazole analogues as tumor hypoxia markers. AB - Three novel nitroimidazole-based thioflavin-T derivatives, N-[4-(benzothiazol-2 yl)phenyl]-3-(4-nitroimidazole-1-yl)propanamide, N-[4-(benzothiazol-2-yl) phenyl] 3-(4-nitroimidazole-1-yl)-N-methylpropanamide and N-[4-(benzothiazol-2-yl)phenyl] 3-(2-nitroimidazole-1-yl) propanamide were synthesized and radiolabeled with iodine-131. Three (131)I-labeled compounds continuously accumulated in hypoxic murine sarcoma S180 cells in vitro but not in aerobic cells. Biodistribution results in mice bearing S180 tumor indicated that the tracers could localize in the tumor and eliminate from it slowly. In contrast, the uptake in other organs (stomach excluded) was little and the clearance was quick. The tumor-to-tissue ratios of three compounds all increased with time. PMID- 15820758 TI - In vivo apoptosis detection with radioiodinated Annexin V in LoVo tumour-bearing mice following Tipifarnib (Zarnestra, R115777) farnesyltransferase inhibitor therapy. AB - In this paper, the use of (123)I-Annexin V for the detection of farnesyltransferase inhibitor (FTI)-induced apoptosis in tumour-bearing athymic mice is described. In vitro binding assays on LoVo cells show time- and dosage dependent (125)I-Annexin V binding upon treatment with Tipifarnib (Zarnestra, R115777), a selective and potent FTI. In vivo experiments using planar gamma scintigraphy on LoVo inoculated mice show a 40% increased (123)I-Annexin V uptake 8 h after a single oral administration of 100 mg/kg Tipifarnib in 20% beta cyclodextrin in 0.1 M HCl, as well as after 3 days of twice daily treatments with the same dose. Ex vivo TUNEL assays, detecting end-stage apoptotic cells, correlate significantly with both in vitro and in vivo results. The percentage of necrosis is also increased by Tipifarnib treatment, but is too low to interfere with the (123)I-Annexin V uptake. It can be concluded that (123)I-Annexin V can be used to monitor Tipifarnib-induced apoptosis in LoVo xenograft tumours in athymic mice. Future applications might include the early prediction of FTI response and the selection of FTI-sensitive patients very shortly after treatment initiation. Subsequently, such patients would greatly benefit from a noninvasive and fast therapy evaluation. PMID- 15820759 TI - MBP-annexin V radiolabeled directly with iodine-124 can be used to image apoptosis in vivo using PET. AB - A noninvasive method of measuring programmed cell death in the tumors of cancer patients using positron-emission tomography (PET) would provide valuable information regarding their response to therapeutic intervention. Our strategy is to radiolabel annexin V, a protein that binds to phosphatidylserine moieties that are translocated to the external leaflet of plasma membranes during apoptosis. We developed a phosphatidylserine-ELISA capable of distinguishing wild type from point mutant annexin V that is known to have a lower phosphatidylserine binding affinity. A maltose-binding protein/annexin V chimera was synthesized and purified with high yield using amylose resin. We showed that it bound to phosphatidylserine in the ELISA as well as to that exposed on apoptotic Jurkat cells; therefore, it was used in the development of a method for radiolabeling annexin V using iodine radionuclides. MBP-annexin V retained its phosphatidylserine binding properties on direct iodination, but at high levels of oxidizing agents (iodogen and chloramine T), its specificity for phosphatidylserine was compromised. (124)I-MBP-annexin V was successfully used to image Fas-mediated hepatic cell death in BDF-1 mice using PET. In conclusion, we have shown that MBP-annexin V and the phosphatidylserine ELISA are useful tools for the development of methods for radiolabeling annexin V for PET imaging. PMID- 15820761 TI - Biological properties of 2'-[18F]fluoroflumazenil for central benzodiazepine receptor imaging. AB - A novel positron emitting agent, 2'-[18F]fluoroflumazenil (fluoroethyl 8-fluoro-5 methyl-6-oxo-5,6-dihydro-4H-benzo-[f]imidazo[1,5-a][1,4]diazepine-3-carboxylate, FFMZ), has been reported for benzodiazepine imaging. In the present study, biological properties of [18F]FFMZ were investigated. Stability tests of [18F]FFMZ in human and rat sera were performed. Biodistribution was investigated in mice and phosphorimages of brains were obtained from rats. A receptor binding assay was performed using rat brain (mixture of cortex and cerebellum) homogenate. A static positron emission tomography (PET) image was obtained from a normal human volunteer. Although [18F]FFMZ was stable in human serum, it was rapidly hydrolyzed in rat serum. The hydrolysis was 39%, 63% and 92% at 10, 30 and 60 min, respectively. According to the biodistribution study in mice, somewhat even distribution (between 2 approximately 3% ID/g) was observed in most organs. Intestinal uptake increased up to 6% ID/g at 1 h due to biliary excretion. Bone uptake slowly increased from 1.5% to 3.5% ID/g at 1 h. High uptakes in the cortex, thalamus and cerebellum, which could be completely blocked by coinjection of cold FMZ, were observed by phosphorimaging study using rats. Determination of Kd value and Bmax using rat brain tissue was performed by Scatchard plotting and found 1.45+/-0.26 nM and 1.08+/-0.03 pmol/mg protein, respectively. The PET image of the normal human volunteer showed high uptake in the following decreasing order: frontal cortex, temporal cortex, occipital cortex, cerebellum, parietal cortex and thalamus. In conclusion, the new FMZ derivative, [18F]FFMZ appears to be a promising PET agent for central benzodiazepine receptor imaging with a convenient labeling procedure and a specific binding property. PMID- 15820760 TI - Preclinical and clinical evaluation of O-[11C]methyl-L-tyrosine for tumor imaging by positron emission tomography. AB - We performed preclinical and clinical studies of O-[11C]methyl-L-tyrosine, a potential tracer for imaging amino acid transport of tumors by positron emission tomography (PET). Examinations of the radiation-absorbed dose by O-[11C]methyl-L tyrosine and the acute toxicity and mutagenicity of O-methyl-L-tyrosine showed suitability of the tracer for clinical use. The whole-body imaging of monkeys and healthy humans by PET showed low uptake of O-[11C]methyl-L-tyrosine in all normal organs except for the urinary track and bladder, suggesting that the O [11C]methyl-L-tyrosine PET has the potential for tumor imaging in the whole-body. Finally, the brain tumor imaging was preliminarily demonstrated. PMID- 15820762 TI - 177Lu-antibody conjugates for single-cell kill of B-lymphoma cells in vitro and for therapy of micrometastases in vivo. AB - Antibodies (Abs) conjugated to 177Lu, a relatively low-energy beta-particle emitter, were evaluated in vitro for their cytotoxic activity and in vivo for their therapeutic activity against disseminated B-cell lymphoma xenografts in SCID mice. 177Lu was compared with other beta-particle emitters ((131)I and 90Y), and also with emitters of low-energy electrons (LEEs, meaning Auger and conversion electrons of < 50 keV). The Abs used reacted with CD20, CD74 or HLA DR, and the target cell was the Raji B lymphoma. Like the other beta-particle emitters, 177Lu was a potent and specific toxic agent in vitro, when conjugated to Abs recognizing high-density antigens. It appeared to be slightly less potent than (131)I per decay, but this difference was relatively small, and would not be a major factor in the selection of the optimal radionuclide for clinical use. The nonspecific toxicity from 177Lu was less than from 90Y, but 177Lu still produced greater nonspecific toxicity in vitro than LEE emitters. The maximum tolerated dose (MTD) of 177Lu-anti-CD74 in SCID mice was 1.81 MBq (49 microCi)/mouse. When this dose was administered on day 5 after tumor inoculation, significant protection was obtained, but considerably less than the protection obtained in previous experiments with LEE emitters (111)In and 67Ga. In conclusion, 177Lu has advantages over other available beta-particle emitters as a therapeutic agent, but its efficacy in the treatment of micrometastases seems to be less than that of LEE emitters, due to greater nonspecific toxicity. This conclusion, however, may not apply to therapy of macroscopic tumors. PMID- 15820763 TI - Technetium labeling of dextran incorporating cysteamine as a ligand. AB - INTRODUCTION: Technetium-99m-labeled dextran is a useful imaging agent for procedures such as angiocardiography and lymphoscintigraphy. To improve the availability of 99mTc-labeled dextran, we designed a cysteamine ligand system for dextran labeling. METHODS: Cysteamine derivatized dextran was synthesized as follows. Dextran was oxidized with sodium periodate, coupled with cysteamine and reduced with sodium borohydride to provide the desired amine ligand. The cysteamine-dextran conjugate was then labeled with reduced 99mTc. Whole-body scintigraphy and biodistribution were examined following injection of the 99mTc labeled cysteamine-conjugated dextran (99mTc-cysteamine-dextran) in ICR mice. Lymphoscintigraphy was performed after intradermal injection of 99mTc-cysteamine dextran in SD rats. RESULTS: The cysteamine-derived dextran was easily labeled with reduced 99mTc in greater than 96% yield. 99mTc-cysteamine-dextran has a higher chelation stability against diethylenetriamine pentaacetic acid (DTPA) than the 99mTc-dextran. Axillary lymph nodes were clearly visible after intradermal injection of 99mTc-cysteamine-dextran in rats. CONCLUSION: These results suggest that 99mTc-cysteamine-dextran is available for lymphoscintigraphy. This methodology could expand the usage of 99mTc-labeled dextran, particularly for diagnostic purposes. PMID- 15820764 TI - Gallium-68-DOTA-albumin as a PET blood-pool marker: experimental evaluation in vivo. AB - Investigations into tumor angiogenesis and antiangiogenic treatment have renewed interest in tumor perfusion. To image tumor blood-pool by PET, suitable tracers are not generally available. In this experimental study, we characterized a 68Ga labeled 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) conjugate of rat serum albumin (68Ga-DOTA-RSA) in vivo using a generator-produced isotope. Biodistribution was determined in ACI rats after intravenous administration of 3 6 MBq of 68Ga-DOTA-RSA. Three ACI rats were imaged over 1 h by dynamic PET after intravenous administration of 15-25 MBq of 68Ga-DOTA-RSA while the blood-pool activity was recorded simultaneously in a closed extracorporeal loop (ECL) between the carotid artery and the jugular vein. Time-activity curves (TACs) were obtained from volume of interest (VOI) analysis and from the ECL data. Stability and metabolites in plasma and urine were analyzed by size exclusion HPLC (SE HPLC) 1 h after intravenous injection of 67Ga-DOTA-RSA. Blood radioactivity decreased by 10% and 18% from 10 to 60 min p.i. by biodistribution and PET or ECL, respectively. Tissue sampling between 10 and 60 min p.i. showed slight increases in the uptake of spleen, myocardium, kidney and skeletal muscle while hepatic accretion remained unchanged. Total urinary excretion after 60 min amounted to 9% of the injected dose. HPLC demonstrated a single urinary metabolite corresponding in size to gallium-labeled DOTA. 68Ga-DOTA-RSA is a blood-pool tracer whose physical and biological half-life is well suited for PET. Our findings support clinical imaging using 68Ga-DOTA-labeled human serum albumin (HSA). The generator-produced label makes 68Ga-DOTA-labeled albumin continuously available even to centers lacking an in-house cyclotron. PMID- 15820765 TI - In vivo evaluation of [(123)I]-3-(4-iodobenzyl)-1,2,3,4-tetrahydro-8 hydroxychromeno[3,4-c]pyridin-5-one: a presumed dopamine D4 receptor ligand for SPECT studies. AB - [(123)I]-3-(4-iodobenzyl)-1,2,3,4-tetrahydro-8-hydroxychromeno[3,4-c]pyridin-5 one ([(123)I]-ITCP), a presumed radioligand for visualization of the dopamine D4 receptor by single photon emission computed tomography, was evaluated in vivo in mice and rabbits. This new radioiodinated tracer exhibited high brain uptake (3.64% injected dose per gram of tissue at 10 min p.i.) in mice. No significant amounts (less than 5%) of labeled metabolites were present in the brain, as demonstrated by a metabolite study. Regional brain distribution in rabbits showed atypical CNS uptake with consistently low values in the cortex and high values in other brain parts including cerebellum. Saturable binding was confirmed by a competition experiment with unlabeled product. Selectivity was assessed by competition experiments with a known dopamine D4 ligand and later with a sigma receptor ligand. Both experiments showed no observable competition. In conclusion, our findings indicate that [(123)I]-ITCP is neither a dopamine D4 receptor ligand nor a sigma receptor ligand. The exact nature of [(123)I]-ITCP binding in the brain remains to be elucidated. PMID- 15820767 TI - Optimizing an online SPE-HPLC method for analysis of (R)-[11C]1-(2-chlorophenyl) N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxamide [(R)-[11C]PK11195] and its metabolites in humans. AB - (R)-[11C]PK11195 is used as a positron emission tomography tracer for activated microglia in several neurological disorders. Quantification of specific binding requires a metabolite-corrected plasma input function. In this study, a high performance liquid chromatography (HPLC) procedure with online solid phase extraction was modified for analyzing (R)-[11C]PK11195 plasma samples, yielding total sample recoveries of more than 98%. When applied to human studies, the use of two HPLC systems enabled analysis of up to seven plasma samples under regular conditions. Online radioactivity detection was compared with offline sample measurements of HPLC profiles. Offline measurements provided the most reliable results especially for late plasma samples. In 10 patients, an average decrease of parent compound from 94.6% at 2.5 min to 45.2% at 1 h after administration was observed. PMID- 15820766 TI - Purification of cyclotron-produced 203Pb for labeling Herceptin. AB - A simple and rapid procedure was developed for the purification of cyclotron produced 203Pb via the 203Tl(d,2n) 203Pb reaction. A Pb(II) selective ion exchange resin, with commercial name Pb Resin from Eichrom Technologies, Inc., was used to purify 203Pb from the cyclotron-irradiated Tl target with excellent recovery of the enriched Tl target material. The purified 203Pb was used to radiolabel the monoclonal antibody Herceptin. The in vitro and in vivo properties of the 203Pb radioimmunoconjugate were evaluated. PMID- 15820768 TI - The mechanisms of differential glucocorticoid and mineralocorticoid action in the brain and peripheral tissues. AB - Mineralocorticoids and glucocorticoids are key categories of adrenocorticosteroid hormones that mediate distinct physiological responses. While the primary role of aldosterone, the major mineralocorticoid, is in regulating sodium homeostasis, the major role of the glucocorticoids is mediating the catabolic response to stress. Over the past two decades, these adrenocorticosteroid hormones have been the subject of considerable attention due to the paradox that despite exerting greatly different physiological effects, they act through very closely related receptors and a common DNA response element. This review will examine the research focused on the mechanisms of selective adrenocorticosteroid action. In general, it has been demonstrated that differential adrenocorticosteroid action is mediated at pre-receptor, receptor, and post-receptor levels, depending on the target tissue and physiological environment. The marked neuroendocrine pathophysiologies resulting from perturbations in this complex system make it imperative that further research into mechanisms of coordination of the three levels of adrenocorticosteroid control be conducted. PMID- 15820769 TI - Methodological issues affecting the determination of plasma matrix metalloproteinase (MMP)-2 and MMP-9 activities. AB - OBJECTIVES: To examine whether a linear relationship exists between the volume of plasma loaded in each gel lane and the gelatinolytic activities of plasma matrix metalloproteinase (pMMP)-2 and pMMP-9, and the influences of plasma storage temperature, fasting, and a variable number of repeated freeze-thaw cycles. DESIGN AND METHODS: Plasma from venous blood collected into EDTA tubes was stored for 1 month at -20 degrees C or at -70 degrees C until assayed for gelatin zymography of pMMP-2 and pMMP-9. The effects of food intake, repeated freeze-thaw cycles, and the gelatinolytic activity produced by serial dilutions of plasma were examined. RESULTS: Storage temperature, fasting, and up to five repeated freeze-thaw cycles had no effect on the activities of pMMPs (all P < 0.05). A linear relationship was found with plasma volumes below 0.75 microL/lane for MMP 2, and from 0.75 microL/lane to 2 microL/lane for MMP-9. CONCLUSIONS: Different volumes of plasma should be studied when pMMP-2 and pMMP-9 activities are studied. Storage temperature, repeated freeze-thaw cycles, and food intake do not affect the activities of pMMPs. PMID- 15820770 TI - Vitamin D-binding protein gene polymorphism association with IA-2 autoantibodies in type 1 diabetes. AB - BACKGROUND: Vitamin D-binding protein (DBP) is the main systemic transporter of 1.25(OH)2D3 and is essential for its cellular endocytosis. There are two known polymorphisms in exon 11 of the DBP gene resulting in amino acid variants: GAT- >GAG substitution replaces aspartic acid by glutamic acid in codon 416; and ACG- >AAG substitution in codon 420 leads to an exchange of threonine for lysine. These DBP variants lead to differences in the affinity for 1.25(OH)2D3. Correlations between DBP alleles and type 1 diabetes have been described in different populations. Therefore, we investigated the polymorphism in codon 416 of the DBP gene for an association with autoimmune markers of type 1 diabetes. DESIGN AND METHODS: The present analysis was a case control study. 110 patients, 68 controls, and 115 first-degree relatives were genotyped for the DBP polymorphism in codon 416. DNA typing of DBP locus was performed by the PCR restriction fragment length polymorphism method (RFLP). RESULTS: The frequencies of the Asp/Glu and Glu/Glu were significantly increased in diabetic subjects with detectable IA-2 antibodies (P < 0.01). On the contrary, the DBP Glu-containing genotype was not accompanied by differences in the prevalence of GAD65 antibodies. These finding supports a role of the vitamin D endocrine system in the autoimmune process of type 1 diabetes. PMID- 15820771 TI - Mutation analysis of the preproghrelin gene: no association with obesity and type 2 diabetes. AB - OBJECTIVE: To investigate the preproghrelin gene for variants and their association with obesity and type 2 diabetes. DESIGN AND METHOD: 82 obese probands were analyzed for mutations using single-strand conformational polymorphism, heteroduplex analyses and sequencing. Association studies were performed in 234 juvenile-onset obese and 323 lean men and in 557 type 2 diabetic and 233 glucose tolerant subjects. RESULTS: We identified two novel variants, 36C > T and IVS3 + 715delC, and 4 known variants, Arg51Gln, Leu72Met, Gln90Leu, and IVS1 + 169G > A. None of the variants showed any significant association with obesity or type 2 diabetes or estimates of glucose and lipid metabolism in glucose tolerant subjects. CONCLUSION: Variation in the preproghrelin gene is not associated with juvenile-onset obesity, type 2 diabetes or related phenotypes among the examined Danish Caucasian subjects. PMID- 15820772 TI - Alpha-1-antitrypsin phenotypes and HLA-B27 typing in uveitis patients in southeast Iran. AB - OBJECTIVES: Uveitis is an eye disease that affects humans worldwide. Inflammation of the uveal tract is termed uveitis. Alpha-1-antitrypsin (AAT) deficiency is one of many factors that may be involved in abnormalities such as liver and lung disease, inflammatory joint diseases, and inflammatory eye diseases. In this study, the role of AAT in uveitis is analyzed. DESIGN AND METHODS: AAT phenotyping and serum-trypsin inhibitory capacity (S-TIC) experiments were performed on 103 patients who were referred to the ALZAHRA eye center in Zahedan (southeast of Iran). The same experiments were performed on 167 people who did not suffer from any eye or systemic diseases and served as a control group. RESULTS: The results revealed that the frequency of M1S, M2S, M1Z, and MV phenotypes were significantly higher in uveitis patients (P < 0.001). There was no difference in AAT phenotype frequencies between various types of uveitis (P = 0.1). CONCLUSION: AAT deficiency appears to be a risk factor for uveitis in southeast Iran. More investigation is needed to establish potential benefits of AAT phenotyping tests and AAT therapy in the diagnosis and treatment of uveitis cases with unclear etiology. PMID- 15820773 TI - Ontogenetic changes in serum S100B in Down syndrome patients. AB - BACKGROUND: It has been shown that Down syndrome (DS) patients have elevated S100B levels in brain tissue. DESIGN: Measurements of S100B were performed in serum samples from 48 DS patients and 42 ostensibly healthy age-matched controls. RESULTS: We observed higher levels of S100B in the DS group than in the control group. Moreover, serum S100B in DS patients was not age-dependent as it is in normal individuals. CONCLUSION: The higher levels of S100B in DS patients may reflect a general and persistent increase in the extracellular space and may be associated with neurodegenerative lesions observed in DS patients. PMID- 15820774 TI - Evidence for the association of synaptotagmin with glutathione S-transferases: implications for a novel function in human breast cancer. AB - OBJECTIVE: To analyze the pattern of changes in GSTs in cancerous and adjacent non-cancerous tissues obtained from breast cancer patients undergoing surgery. DESIGN AND METHODS: Cytosolic GST purification, assay of GST, protein expression levels, and GST-synaptotagmin association were analyzed using standard biochemical techniques like GSH-affinity purification, spectrophotometry, SDS PAGE, Western blots, and matrix-assisted laser desorption and ionization-time of flight (MALDI-TOF). RESULTS: GST activity in cancerous tissues (0.26 U/mg protein) was significantly higher (P < 0.05) as compared to those from adjacent non-cancerous tissues (0.14 U/mg protein) of breast cancer patients. Further analysis of GST subunits on SDS-PAGE and Western blots using class-specific GST antibodies revealed significant elevation in GST-pi levels in cancer tissues with no appreciable changes in GST-alpha and GST-mu. Along with the elevation of GST pi levels, high molecular weight proteins (approximately 70 kDa) cross reacting with GST antibodies were detected only in surgically resected tumor biopsies but not in the non-cancerous tissues adjacent to the tumor. Based on MALDI-TOF analysis, the high molecular weight band was identified as synaptotagmin V bound to GST-M1 with 47% sequence coverage after processing on an MS-FIT search engine. CONCLUSIONS: Our results suggest a novel putative functional role for the GST synaptotagmin complex in human breast cancers. As this association of GST M1 synaptotagmin was not seen in adjacent non-cancerous tissues, this can be used as a marker for breast cancers. PMID- 15820775 TI - Carotenoids, retinol and tocopherols in blood: comparability between serum and plasma (Li-heparin) values. AB - OBJECTIVE: To assess the comparability of concentrations of retinol, alpha- and gamma-tocopherols and individual carotenoids in serum and (Li-hep) plasma over a wide range of concentrations. MATERIAL AND METHODS: One hundred sixty-six pairs of samples (serum and lithium-heparin plasma) were analyzed by a quality controlled HPLC method. Means and 95% confidence intervals, differences, interchangeability and the degree of agreement (Bland-Altman plot) were calculated. RESULTS: Distribution of all analytes in the two matrices are comparable and interchangeable with minor quantitative adjustments. Within the range of concentrations assessed, the degree of agreement was high, although some differences were observed for minor components with greater analytical imprecision. CONCLUSIONS: The results indicate an acceptable degree of agreement using either of the two matrices for the analytes assessed except possibly for minor blood components. For retinol and alpha-tocopherol, the comparability and interchangeability of results below the cut-off points for inadequacy need further confirmation. PMID- 15820776 TI - Measurement of intracellular vitamin C levels in human lymphocytes by reverse phase high performance liquid chromatography (HPLC). AB - OBJECTIVES: Vitamin C plays an active role in many important metabolic processes, such as collagen formation and the prevention of bleeding. Although overt scurvy is now rare, there is evidence that subclinical vitamin C deficiency is still quite common. Serum and plasma vitamin C measurements do not correlate well with tissue levels while lymphocyte vitamin C levels provide the most accurate assessment of the true status of vitamin C stores and are not affected acutely by circadian rhythm or dietary changes. We report a specific and reproducible reverse phase high performance liquid chromatographic method (HPLC) for the quantification of vitamin C in lymphocytes. METHODS: Reverse phase HPLC with a UV detection system was used. National Committee for Clinical Laboratory Standards (NCCLS) guidelines were followed for evaluation. Sample stability testing for lymphocyte vitamin C was performed for a period of 24 h at room temperature and 4 degrees C. Lymphocyte vitamin C levels were measured in 51 children. RESULTS: Lymphocyte vitamin C measurement with HPLC revealed very good analytical sensitivity with a 1.42 microg/10(8) lymphocyte lower limit of detection on repeated testing. An external standard curve was used for quantification, which showed a linear range of 1.25-100 microg/10(8) lymphocyte with a correlation coefficient of 0.989. Precision studies showed an inter-assay repeatability coefficient of variance (CV) between 0.25-9.98% and a within-assay coefficient of variance between 1.2-12.49%. The inter-assay CV for a period of 20 days was less than 10% for concentrations equal to or less than 1.42 microg/10(8) lymphocytes and less than 5.5% for concentrations between 5-100 microg/10(8) lymphocytes. Vitamin C was most stable at 4 degrees C, with a 0.31% decrease after 3 h and 2.35% after 4 h. At room temperature, vitamin C loss was more significant, with losses of 8.44% and 15.6% at 3 and 4 h, respectively, at a concentration of 29.9 microg/10(8) lymphocytes. CONCLUSIONS: The proposed HPLC method offers a reliable and reproducible technique for the quantification of intracellular vitamin C. Lymphocyte samples can be rapidly prepared and represent a more homogeneous tissue sample source for intracellular vitamin C measurement as compared to serum. To ensure stability, lymphocyte lysates should be prepared and stored at or below -20 degrees C within 2 h of blood collection. PMID- 15820777 TI - Magnesium sulfate effect on erythrocyte membranes of asphyxiated newborns. AB - OBJECTIVES: Magnesium sulfate has been recognized as a neuroprotective agent against hypoxia-ischemia, mainly by the protection from the excitotoxicity associated with increased glutamate concentration. However, the mechanism of MgSO4 action is not fully understood and is considerably controversial. DESIGN AND METHODS: During the 2 first hours of life, the asphyxiated full-term newborns were treated intravenously with one dose of MgSO4 250 mg/kg body weight. At birth, after 6 and 48 h of life the activity of ATP-dependent enzymes in erythrocyte membranes: Mg2+-ATPase, Ca2+-ATPase, protein kinases A and C, were determined. Using monoclonal antibodies, the band 3 and its phosphotyrosine level were also assayed. RESULTS: The time-dependent decrease of Ca2+-ATPase activity was detected in untreated newborns, whereas MgSO4 prevented this reduction. After 48 h, protein kinases activities differed in MgSO4-treated and untreated groups. Magnesium therapy increased the amount of band 3 and diminished proteolytic degradation of this protein. CONCLUSION: Our results demonstrated, for the first time, that magnesium sulfate treatment significantly altered the activities of some important enzymes in erythrocyte membrane from asphyxiated newborns. It also reduced the post-asphyxial damages of membrane compounds. These data may partly explain the molecular mechanisms of MgSO4 action in asphyxiated newborns. PMID- 15820778 TI - Human heterophilic antibodies display specificity for murine IgG subclasses. AB - OBJECTIVES: The study investigated heterophilic antibodies: the human immunoglobulin classes involved and their specificity for different murine IgG subclasses. DESIGN AND METHODS: Using immunofluorometric assays for human IgA, IgM and IgG binding murine IgG1, we analyzed 173 samples displaying positive interference and 97 negative control samples from a previous study. We also set up assays for heterophilic antibody interference using Mabs from different murine IgG subclasses. Three Mabs each of murine IgG1, IgG2a and IgG2b subclasses, one murine IgG3 Mab and one rat Mab were used. RESULTS: Elevated levels of human murine IgG1-binding immunoglobulins of IgM class only were found in 40% of interference-positive samples, human IgG only in 1.7%, and human IgA only in 2.3% of the samples. Both elevated human IgG and IgM classes were found in 3.5% of the samples, IgA and IgM in 4.0%, and finally, all three immunoglobulin classes in 1.7% of the samples. Eighty percent of interference positive samples showed heterophilic assay interference for at least one murine IgG1 Mab, 35% for IgG2a, 66% for IgG2b, 52% for IgG3a and 17% for the rat Mab. CONCLUSIONS: Heterophilic antibody interference is mainly caused by IgM class human antibodies with a marked murine IgG subclass specificity. Combining assay antibodies from different murine IgG subclasses may reduce interference in immunoassays. PMID- 15820779 TI - Analytical variability of the Fibrotest proteins. AB - OBJECTIVES: The analytical variability of the Fibrotest (FT) parameters raises the issue of the test's reliability for routine use. Whereas standardization has been proposed by the International Federation of Clinical Chemistry (IFCC) for specific proteins, few data are available concerning the actual transferability of the FT proteins, i.e. haptoglobin, apolipoprotein A1 and alpha2 macroglobulin. The aim of this study was to evaluate the analytical variability of the FT proteins. DESIGN AND METHODS: During the FIBROPACA study, we evaluated 112 sera from patients with hepatitis C infection who underwent liver biopsy. We compared measurements of haptoglobin, apolipoprotein A1 and alpha2 macroglobulin by the autoanalyzers Immage (Beckman-Coulter) and the FT reference BNProspec (Dade Behring). RESULTS: Optimal concordance was found for haptoglobin (correlation: y = 1.05x -0.09; correlation coefficient = 0.98). However, apolipoprotein A1 as determined with Immage was globally 12% lower than with BNProspec (correlation: y = 0.88x -0.05; correlation coefficient = 0.91) and alpha2 macroglobulin values were 40% greater with Immage than with BNProspec (correlation: y = 1.40x -0.46; correlation coefficient = 0.96). CONCLUSIONS: Inter-technique analytical variability of the Fibrotest parameters remains a major issue. After IFCC standardization of specific proteins, some discrepancies remain for alpha2 macroglobulin and, to a lesser extent, for apolipoprotein A1. National and international quality control programs would be useful to monitor analytical performance of protein assays. PMID- 15820780 TI - Problems with the estimation of urine protein by automated assays. AB - OBJECTIVES: Most clinical laboratories replaced their manual precipitation techniques for the determination of urinary protein with automated dye binding assays or benzethonium chloride-turbidimetric assays. Few studies have validated these assays for the measurement of urinary proteins in the normal range. DESIGN AND METHODS: This study compares four automated assays for the measurement of urinary protein to a manual Ponceau S/TCA precipitation assay. We evaluated the linearity, the precision, the analytical sensitivity, the accuracy and the recovery of different proteins for each assay. RESULTS: All assays showed good linearity with the theoretical concentration of albumin present in the sample. The coefficient of variation was below 10% at a concentration of 0.142 g/L. However, the manual Ponceau S/TCA assay demonstrated superior analytical sensitivity. Accuracy determinations showed a variable positive bias and poor correlations at concentrations below 0.1 g/L when compared to the Ponceau S/TCA assay. Small molecular weight peptides particularly affected the pyrogallol red assays but other urinary components also interfered with the automated assays. CONCLUSIONS: Most automated assays show high imprecision and poor accuracy for the measurement of urinary protein in the normal range. The Ponceau S/TCA offers a precise and accurate manual alternative to these automated assays. PMID- 15820781 TI - Serum xylosyltransferase I activity, the new biochemical fibrosis marker, is not affected by renal insufficiency. AB - OBJECTIVES: Serum xylosyltransferase I (XT-I) is a marker for the determination of tissue remodeling in systemic sclerosis. Here, we investigated whether renal insufficiency affects XT-I levels in blood. METHODS: We measured serum XT-I activity in 236 patients with different serum creatinine levels. RESULTS: XT-I activities in cohorts with increased creatinine levels were not significantly altered compared to controls. CONCLUSIONS: Serum XT-I activity is applicable as a fibrosis marker independent from renal function. PMID- 15820782 TI - Association of the PIM3 allele of the alpha-1-antitrypsin gene with chronic obstructive pulmonary disease. AB - OBJECTIVE: The study investigated the association of genetic polymorphism of the alpha1AT gene with COPD. DESIGN AND METHODS: The mutations and polymorphism of alpha1AT gene were investigated by DNA sequence analysis using polymerase chain reaction. RESULTS: The frequency of the PIM3 allele in COPD patients was found to be significantly higher than the controls (P < 0.0001). Five SNPs, including a novel SNP (24_25insA), were observed near the junction of exon-intron I. The occurrence of these SNPs didn't show any association with COPD. However, the PIM3 allele of the alpha1AT gene was found to be associated with COPD. CONCLUSION: The PIM3 allele of the alpha1AT gene is found to have an association with the pathogenesis of COPD in the Indian population. PMID- 15820783 TI - Palmitoyl Protein Thioesterase1 (PPT1) and Tripeptidyl Peptidase-I (TPP-I) are expressed in the human saliva. A reliable and non-invasive source for the diagnosis of infantile (CLN1) and late infantile (CLN2) neuronal ceroid lipofuscinoses. PMID- 15820784 TI - Evidence-based investigations and treatments of recurrent pregnancy loss. AB - OBJECTIVE: To give an overview of currently used investigations and treatments offered to women with recurrent pregnancy loss (RPL) and, from an evidence-based point of view, to evaluate the usefulness of these interventions. DESIGN: Ten experts on epidemiologic, genetic, anatomic, endocrinologic, thrombophilic, immunologic, and immunogenetic aspects of RPL discussed methodologic problems threatening the validity of research in RPL during and after an international workshop on the evidence-based management of RPL. CONCLUSION(S): Most RPL patients have several risk factors for miscarriage, and an extensive investigation for all major factors should always be undertaken. There is an urgent need for agreement concerning the thresholds for detecting what is normal and abnormal, irrespective of whether laboratory tests or uterine abnormalities are concerned. A series of lifestyle factors should be reported in future studies of RPL because they might modify the effect of laboratory or anatomic risk factors. More and larger randomized controlled trials, including trials of surgical procedures, are urgently needed, and to achieve this objective multiple centers have to collaborate. Current meta-analyses evaluating the efficacy of treatments of RPL are generally pooling very heterogeneous patient populations and treatments. It is recommended that future meta-analyses look at subsets of patients and treatment protocols that are more combinable. PMID- 15820785 TI - Changing terminology in reproductive medicine: logical but not simple. AB - Although terminology in reproductive medicine is currently imprecise, it will not be easy to develop a consensus for a new approach to describing couples with difficulty achieving pregnancy. PMID- 15820786 TI - Infertility classification: a work in progress sounds like SOAP revisited. AB - Documentation and communication regarding infertility evaluation and management continue to be a work in progress. Grading systems focused on the degree of infertility and the associated prognosis are a step in the right direction. A subjective, objective, assessment, plan (SOAP) approach is a logical way to provide a detailed method of communication and documentation when dealing with infertile couples. PMID- 15820787 TI - Fertility speak: language and the patient. AB - As the science of reproductive medicine continues to sprint ahead, patients and doctors are still wrestling with the increasingly imprecise and irrelevant terminology of the field. Further, the blurred boundaries between the clinical and social meanings of words such as "infertility" leave patients grappling with the often pejorative connotations associated with the terms, underscoring the need for the development of a socially neutral, clinically relevant, easily understood language of infertility. PMID- 15820788 TI - Research on infertility: which definition should we use? AB - OBJECTIVE: Different definitions of infertility are used in clinical practice as well as in epidemiological and demographic research. This study assessed whether the definition makes a difference for estimates of the prevalence and sociodemographic differentials of infertility and whether one definition would be applicable in both research and clinical practice. DESIGN: Cross-sectional study. SETTING: Moshi town in northern Tanzania. PATIENT(S): Community-based sample of 1,125 women, ages 20 to 44 years, in first union. MAIN OUTCOME MEASURE(S): Six definitions of infertility. RESULT(S): Similar levels and sociodemographic characteristics of infertile women were obtained from asking the question "How long have you tried to get pregnant?" and from secondary data collected in a birth history that included date of marriage, date of last birth, current contraceptive use, and whether the woman wants another child. CONCLUSION(S): The infertility definition made a difference. The World Health Organization definition based on 24 months of trying to get pregnant is recommended as the definition that is useful in clinical practice and research among different disciplines. PMID- 15820789 TI - Reanalysis of the Women's Health Initiative oral contraceptive data reveals no evidence of delayed cardiovascular benefit. AB - Reanalysis of the Women's Health Initiative (WHI) data regarding prior oral contraceptive use and the effect on cardiovascular disease reveals no evidence of a delayed benefit or harm. The WHI database was not designed to answer this question. PMID- 15820790 TI - Duplicate editorial on duplicate publication. AB - The authors define and discuss the various forms taken by duplicate publications, and provide suggested remedies to help authors, editors, reviewers, and readers avoid this form of internal plagiarism. PMID- 15820791 TI - Stress reactivity and family relationships in the development and treatment of endometriosis. AB - OBJECTIVE: To examine stress physiology and facts of family functioning associated with development and course of endometriosis symptoms. DESIGN: Clinical case data and literature review. SETTING: Private practice in hospital affiliated medical office. PATIENT(S): One woman with endometriosis, her parents, and a nonsymptomatic volunteer. INTERVENTION(S): Measures of physiologic and neural reactivity with biofeedback and neurofeedback equipment during family history interview and while sitting quietly. MAIN OUTCOME MEASURE(S): Electroencephalography (EEG), digital skin temperature (DST), electrodermal response (EDR), and electromyography (EMG); facts of family history; contact with family. RESULT(S): Endometriosis symptoms were associated with DST, EDR, EMG, and EEG measures indicating prolonged stress reactions for the symptomatic woman and her parents. Facts of family history and relationships for three generations set the stage for stress reactions. Striking differences are evident in the physiology, family history, and contact with family of the nonsymptomatic woman. CONCLUSION(S): Differences warrant further study, a larger sample, and additional measures using hormone assay to establish connections between stress reactions in the family, endometriosis symptoms, and response to treatment. Further research will document changes in physiology and in symptoms that accompany interruption of stress reactions with self-regulation training and family systems psychotherapy. PMID- 15820792 TI - Elevated levels of macrophage migration inhibitory factor in the peripheral blood of women with endometriosis. AB - OBJECTIVE: To evaluate the concentrations of macrophage migration inhibitory factor (MIF) in the peripheral blood of normal women and patients with endometriosis. DESIGN: Retrospective study using ELISA to measure peripheral blood MIF. SETTING: Gynecology clinic and human reproduction research laboratory. PATIENT(S): Thirty-eight normal women and 55 women with endometriosis. INTERVENTION(S): Peripheral blood samples were obtained a few days before laparoscopy. MAIN OUTCOME MEASURE(S): The MIF concentrations in blood serum. RESULT(S): This current study showed a 364% increase in MIF concentrations in women with endometriosis as compared to normal women. A significant increase was seen in endometriosis stages I-II, but a more marked increase was observed in the more advanced stages of the disease (III-IV). Both fertile and infertile women with endometriosis had higher levels of MIF than normal controls, but the difference was more significant in infertile women with endometriosis. Women with endometriosis with no pelvic pain had higher levels of MIF than normal controls, but a more significant increase in MIF levels was observed in women with endometriosis reporting pelvic pain. CONCLUSION(S): This study showed a marked increase in MIF concentrations in the peripheral blood of women with endometriosis and a relationship with disease progress, and suggests that MIF may be involved in endometriosis-related pain and infertility. PMID- 15820793 TI - Changes in cytokine levels of patients with ovarian endometriosis after treatment with gonadotropin-releasing hormone analogue, ultrasound-guided drainage, and intracystic recombinant interleukin-2. AB - OBJECTIVE: To determine the changes in cytokine levels from women with endometriomas who are treated with recombinant interleukin-2 after ultrasound guided cyst aspiration, and to relate these changes to the clinical results observed in these patients. DESIGN: A double-blind randomized controlled trial. SETTING: University hospital. PATIENT(S): Twenty-four women with endometriosis related symptoms and endometriomas. INTERVENTION(S): Endometriomas in women receiving GnRH analogues and undergoing transvaginal ultrasound-guided cyst aspiration were injected with dextrose that did or did not contain recombinant interleukin-2 (IL-2). Serum samples were collected before and after treatment. MAIN OUTCOME MEASURE(S): Serum samples were analyzed by enzyme immunoassay to determine the levels of IL-1beta, IL-2, IL-6, IL-8, IL-10, IL-12, IL-13, and IL 17. RESULT(S): The cytokine levels after treatment with GnRH analogues and recombinant IL-2 were similar to the initial levels. The patients receiving GnRH analogues without IL-2 had higher IL-1, IL-2, IL-8, and IL-13 levels. Good clinical results were observed in 90% of the patients in the first group but in only 30% of the second one. CONCLUSION(S): Administration of recombinant IL-2 intracystically decreases cytokine production in women with endometriomas. These results have important implications for the design of future therapies based on immunomodulation, such as using higher or repeated doses of recombinant IL-2 in the cysts. PMID- 15820794 TI - Elevated interleukin-16 levels in the peritoneal fluid of women with endometriosis may be a mechanism for inflammatory reactions associated with endometriosis. AB - OBJECTIVE: To investigate the role of interleukin (IL)-16 in peritoneal fluid in the pathogenesis of endometriosis. DESIGN: Comparative and laboratory study. SETTING: University of Tokyo Hospital. PATIENT(S): Peritoneal fluids (PFs) were collected from women without endometriosis (n = 34) and with endometriosis (stages I/II, n = 30; stages III/IV, n = 58). Peritoneal fluid mononuclear cells (PFMCs) were collected from six women. INTERVENTION(S): The PFs were collected; PFMCs were isolated and cultured with or without recombinant human (rh) IL-16. MAIN OUTCOME MEASURE(S): Concentrations of IL-16 in PFs were measured by enzyme linked immunosorbent assay (ELISA). Concentrations of IL-6, tumor necrosis factor alpha (TNF-alpha), and IL-1beta in culture media of PFMCs were determined by ELISA. RESULT(S): The IL-16 concentrations in the PF of women with advanced endometriosis (stages III/IV) (330 pg/mL, 231-501; median, interquartile range) were significantly higher (P=.0016) than those without endometriosis (229 pg/mL, 174-311). The PFMCs cultured with rhIL-16 released an increased amount of IL-6, TNF-alpha and IL-1beta, which was a 1.17-, 1.33-, and 1.54-fold increase, respectively, over that in the control culture. CONCLUSION(S): The present study indicates that IL-16 in PF may play a role in the pathogenesis of endometriosis by initiating or sustaining inflammatory responses in the peritoneal cavity. PMID- 15820795 TI - The serum follicle-stimulating hormone-to-luteinizing hormone ratio at the start of stimulation with gonadotropins after pituitary down-regulation is inversely correlated with a mature oocyte yield and can predict "low responders". AB - OBJECTIVE: The aim of this study was to investigate the relationship of serum FSH and LH levels at the commencement of stimulation to ovarian follicular development in women undergoing pituitary down-regulation and controlled ovarian hyperstimulation with gonadotropins in IVF or intracytoplasmic sperm injection (ICSI) cycles. DESIGN: Retrospective analysis. SETTING: An IVF program in a tertiary medical center. PATIENT(S): A total of 245 women proven to be pituitary down-regulated by their serum E(2) levels. INTERVENTION(S): Patients treated with a GnRH agonist and FSH and hMG underwent assisted reproductive technique (ART). MAIN OUTCOME MEASURE(S): Mature oocyte yield, pregnancy rate (PR), and live birth rate. RESULT(S): The serum FSH levels and the FSH-to-LH ratio at the commencement of gonadotropin stimulation were inversely correlated to the number of mature oocytes (r = -0.193 and r = -0.224, respectively). When assessed with receiver operating characteristic (ROC) analysis, there was statistically significant ability for the FSH/LH ratio to differentiate between the "poor response" cycles (with mature oocyte yield < or =4) and the normal response cycles. Using the cutoff value derived from ROC analysis, cycles with the FSH-to-LH ratio > or =3 produced less mature oocytes (8.25 vs. 11.74), lower peak E(2) levels (1,975.3 pg/mL vs. 3,324.8 pg/mL), and higher percentage of poor ovarian response cycles (32.5% vs. 14.3%). CONCLUSION(S): The serum FSH-to-LH ratio at the start of gonadotropin stimulation after pituitary down-regulation provided a practical method for early prediction of mature oocyte yield. PMID- 15820797 TI - Efficacy of in vitro fertilization after chemotherapy. AB - OBJECTIVE: To evaluate if in vitro fertilization (IVF) with embryo cryopreservation can be proposed to patients immediately after one or two regimens of chemotherapy. DESIGN: Retrospective study. SETTING: Academic research center and IVF unit. PATIENT(S): Eleven young patients diagnosed with cancer between September 1999 and April 2003 who wanted to preserve their fertility via IVF. INTERVENTION(S): Stimulation and IVF before or soon after chemotherapy treatment. MAIN OUTCOME MEASURE(S): The number and quality of embryos obtained after stimulation in cancer patients undergoing IVF before or soon after chemotherapeutic treatment. RESULT(S): Four patients underwent IVF in the interval between two regimens of chemotherapy. Two of them had no follicular development; one underwent follicular puncture but no oocytes were retrieved; and, in one, six oocytes were harvested but only one good quality embryo was obtained. In the seven patients who underwent IVF before starting chemotherapy, between 4 and 11 embryos were obtained per patient, the majority being good quality embryos. CONCLUSION(S): Because the efficacy of IVF is dramatically reduced after even one round of chemotherapy, IVF should be performed before chemotherapy. For those who require immediate chemotherapy, ovarian tissue cryopreservation and/or oocyte cryopreservation could be used before treatment. PMID- 15820796 TI - Early cessation of triptorelin in in vitro fertilization: a double-blind, randomized study. AB - OBJECTIVE: To compare the efficacy of two early cessation protocols of triptorelin treatment in controlled ovarian hyperstimulation with the conventional long protocol in in vitro fertilization/intracytoplasmic sperm injection. DESIGN: A double-blind, randomized, multicenter study. SETTING: Three Dutch hospitals. PATIENT(S): One hundred seventy-eight women randomized to one of three treatment groups at the start of stimulation. INTERVENTION(S): Midluteally started triptorelin administration was continued until the first day of hMG treatment (group S), or up to and including the fourth day of hMG treatment (group M) or the day of hCG injection (group L). MAIN OUTCOME MEASURE(S): Occurrence of a premature LH surge. RESULT(S): One premature LH surge was observed in group M but not in groups S and L. Both early cessation protocols (S and M) are at least as effective as the long protocol (L) with regard to the number of oocytes (11.1 and 10.3 vs. 9.3), number of embryos (7.3 and 6.5 vs. 5.5), and ongoing pregnancy rate (28% and 24% vs. 21%). CONCLUSION(S): Early cessation of triptorelin on day 1 of hMG treatment in a midluteally started IVF protocol is as effective as the traditional long protocol in preventing a premature LH surge and results in similar fertility effects. PMID- 15820798 TI - Vascular endothelial growth factor and inhibin A in follicular fluid of infertile patients who underwent in vitro fertilization with a gonadotropin-releasing hormone antagonist. AB - OBJECTIVE: To investigate the role of a gonadotropin-releasing hormone (GnRH) antagonist, minimal stimulation protocol, in the follicular fluid by measuring vascular endothelial growth factor (VEGF) and inhibin A. DESIGN: A cross sectional prospective study. SETTING: Academic hospital. PATIENT(S): Seventy infertile patients submitted to in vitro fertilization (IVF). INTERVENTION(S): Patients were divided into two groups: group 1 (study) included 30 infertile patients subjected to IVF with a GnRH antagonist (minimal stimulation protocol); group 2 (control) included 40 infertile women who underwent natural-cycle IVF. MAIN OUTCOME MEASURE(S): Follicular fluid VEGF and inhibin A measurements. RESULT(S): The groups were comparable in terms of age, body mass index (BMI), and infertility characteristics. Moreover, follicular fluid VEGF and inhibin A concentrations (medians) were, respectively, 776 pg/ml (95% confidence interval [CI]: 775-1483) and 3,115 pg/mL (95% CI: 1,349-2,502) for group 1; 1,187.50 pg/mL (95% CI: 1,020-1,560) and 3,123.00 pg/mL (95% CI: 1,888-2,735) for group 2 (P>.05). CONCLUSION(S): We demonstrated that GnRH antagonist administration in infertile patients undergoing IVF did not alter the follicular fluid content of VEGF and inhibin A, and, probably, maturation and quality of oocytes as well. These results demonstrated the usefulness and safety of this drug on controlled ovarian stimulation (COS) protocols. PMID- 15820799 TI - Impact of ovarian endometrioma on oocytes and pregnancy outcome in in vitro fertilization. AB - OBJECTIVE: To evaluate the effect of endometriosis and the presence of an ovarian endometrioma on outcomes of conventional in vitro fertilization (IVF). DESIGN: Retrospective study. SETTING: Reproductive Unit, Department of Obstetrics and Gynecology, Tokai University Hospital. PATIENT(S): Group A: 80 cycles with ovarian endometriomas; group B: 248 cycles with endometriosis but without endometrial cysts at the time of oocyte retrieval; group C: 283 cycles undergoing IVF because of tubal factor without endometriosis. INTERVENTION(S): All conventional in vitro fertilization-embryo transfer (IVF-ET) with previous diagnostic laparoscopy. An endometrioma was diagnosed by direct aspiration at the time of oocyte retrieval. MAIN OUTCOME MEASURE(S): Retrieved number of oocytes, fertilization rate, embryo quality, implantation rate, pregnancy rate, and live birth rate for all cases. In group A, the number of retrieved oocytes in relation to the volume of the endometrioma and affected laterality. RESULT(S): Fewer oocytes were retrieved from groups A and B than from group C (P<.005). The number of retrieved oocytes was not dependent on the volume of endometrial cyst(s). Fertilization rates were similar among the groups. Group A had slightly but not significantly higher rates of morphologically good-quality embryos (group A: 67.2%, group B: 63.0%, group C: 58.1%), implantation (group A: 14.1%, group B: 11.7%, group C: 11.3%), and pregnancy (group A: 25.3%, group B: 22.3%, group C: 23.9%). CONCLUSION(S): Endometriosis affects oocyte number but not embryo quality or pregnancy outcome, irrespective of the presence of an ovarian endometrioma. PMID- 15820800 TI - Parental attitudes toward disclosure of the mode of conception to their child conceived by in vitro fertilization. AB - OBJECTIVE: To survey the level of disclosure of conception method within families of children conceived by conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and to examine the factors that might influence parental attitudes and plans for disclosure. DESIGN: In-depth questionnaire. SETTING: Participants recruited through fertility clinics in the United Kingdom. PATIENT(S): Parents of children aged 5 to 6 years conceived by IVF/ICSI (n = 181; 51% survey response rate). INTERVENTION(S): Mothers and fathers of IVF/ICSI-conceived children were sent questionnaires to complete and return in a postage-paid envelope. MAIN OUTCOME MEASURE(S): Responses to the questionnaire. RESULT(S): Most parents had told somebody about their child's method of conception, mostly close friends and family. Fewer (26% of mothers, 17% of fathers) had already discussed the child's mode of conception with their child. Fifty-eight percent of mothers and 57% of fathers intended to tell their child at some point. Sixteen percent of mothers and 21% of fathers were undecided. Four percent of fathers never wanted to discuss the subject with their child. Children were more likely to be told if conception was ICSI, rather than conventional IVF, and if an only child. Twenty-nine percent of undecided fathers and 36% of undecided mothers stated that they would tell their child if appropriate, child-friendly explanatory literature were available. CONCLUSION(S): The majority of parents wish to tell their child about their conception by IVF/ICSI at some point but are unsure as to the most appropriate timing and method of disclosure. Fertility clinics might have a role to play in providing the necessary support. Child-friendly literature might be helpful. PMID- 15820801 TI - The diagnosis of azoospermia depends on the force of centrifugation. AB - OBJECTIVE: To determine the centrifugal force required to pellet sperm. DESIGN: Prospective, in vitro study. SETTING: Tertiary referral center. PATIENT(S): Men undergoing semen processing or postvasectomy semen analyses. INTERVENTION(S): In Phase I, postvasectomy semen samples were centrifuged at 600 x g, and the pellets were examined. Supernatants from samples with no visible sperm underwent repeat centrifugation at 1,000 x g; the supernatants were removed and centrifuged at 3,000 x g. Pellets from both centrifugations were examined. Phase II examined nonazoospermic semen that was divided into 3 aliquots and centrifuged at 500, 1,000, and 3,000 x g. The supernatants were examined for the presence of sperm. MAIN OUTCOME MEASURE(S): Sperm presence in centrifuged pellets or seminal supernatant. RESULT(S): Phase I: After centrifugation at 1,000 x g and 3,000 x g, sperm were noted in 12% and 0% of samples, respectively. Phase II: Sperm were noted in the supernatant in 100% of samples subjected to 500 x g and 1,000 x g, and in 92% of samples subjected to 3,000 x g centrifugation. CONCLUSION(S): If sperm are in the seminal plasma, they will also be present in the pellet following centrifugation at a force of 1,000 x g or greater. Semen samples that appear azoospermic upon initial wet mount microscopy should be centrifuged at a minimum of 1,000 x g for 15 minutes. PMID- 15820802 TI - A histomorphometric and cytogenetic study of testis from men 29-102 years old. AB - OBJECTIVE: To evaluate the influence of aging on testicular histology and the aneuploidy rate in testis postmeiotic cells. DESIGN: Comparison between older men and younger men. SETTING: Deceased donors and patients who requested assisted reproductive technology (ART). PATIENT(S): Thirty-six older men (61-102 years old) and 10 younger men (29-40 years old). INTERVENTION(S): Testes were obtained from 35 deceased donors, and testicular biopsies were obtained from 11 patients who requested ART. MAIN OUTCOME MEASURE(S): Histomorphometry of testis and fluorescent in situ hybridization (FISH), with a three-probe set X, Y, and 18. RESULT(S): The histomorphometric study showed a thickening of the basal membrane when spermatogenesis was arrested. The number of germinal and Sertoli cells decreased as age increased. The rate of aneuploidy of postmeiotic cells was 1.1% for the control group, 1.29% for older subjects with preserved spermatogenesis, and 14.28% for the subjects with an arrested spermiogenesis. Only this last figure was higher than the control group. CONCLUSION(S): The rate of aneuploidy in older subjects (61-95 years old) with preserved spermatogenesis was not statistically different from that found in the control group; it was increased in older subjects with arrested spermatogenesis. PMID- 15820803 TI - Relationship between reactive oxygen species production and lipid peroxidation in human sperm suspensions and their association with sperm function. AB - OBJECTIVES: To investigate if reactive oxygen species (ROS) production by human sperm suspensions could predict oxidative stress manifest as lipid peroxidation and if lipid peroxidation was a useful predictor of human sperm function. DESIGN: Laboratory study. SETTING: Research laboratory associated with reproductive medicine clinic. PATIENT(S): Eighty-three couples under evaluation for IVF donated the surplus from a diagnostic semen sample. INTERVENTION(S): Cells were separated from seminal plasma on a 30% Percoll gradient. MAIN OUTCOME MEASURE(S): Reactive oxygen species production, lipid peroxidation, sperm motility measured by computer-assisted semen analysis and ability of the sperm to acrosome react. RESULT(S): Forty-four samples exhibited ROS production < or =20 000 relative light units (rlu)/s/10(8) sperm and lipid peroxidation < or =40 nmol malondialdehyde/10(8) sperm. The remainder fell into two distinct groups; both exhibited a linear relationship between lipid peroxidation and ROS production but of markedly differing slopes. Abstinence time had no effect. Lipid peroxidation was a good predictor of sperm motility and their ability to acrosome react. CONCLUSION(S): Reactive oxygen species production in sperm suspensions is an index of overall oxidative stress experienced by sperm, but sperm divide into two classes of ability to resist oxidative stress. Lipid peroxidation should be investigated as a clinical test of sperm function. PMID- 15820804 TI - Molecular profiling of polycystic ovaries for markers of cell invasion and matrix turnover. AB - OBJECTIVE: To study gene expression profiles of connective tissue components in polycystic ovaries using complementary deoxyribonucleic acid (cDNA) array technology. DESIGN: Descriptive study of normal and polycystic human ovarian biopsy samples analyzed by cDNA array hybridizations. SETTING: Experimental laboratory research. PATIENT(S): Eight women with polycystic ovary syndrome (PCOS) and two normally cycling women treated with electrocauterization and hysterectomy, respectively. INTERVENTIONS: Ovarian biopsy samples. MAIN OUTCOME MEASURE(S): Expression levels of 588 genes involved in cellular invasion, extracellular matrix (ECM) turnover, and cell-ECM interactions in polycystic ovaries. RESULT(S): A majority of the 30 genes down-regulated in PCOS ovaries represented those related to cell adhesion and motility, as well as angiogenesis, followed by regulators of cell cycle and growth. The 14 up-regulated genes represented those regulating cell fate and development, growth factors, cytokines, chemokines, and cell-cell interactions. Of the 44 transcripts exhibiting marked changes in the cDNA array analysis, only one - proliferating cell nuclear antigen messenger ribonucleic acid (PCNA mRNA) - was systematically down-regulated; 2 transcripts, for CDC27HS protein and CD9 antigen, were down regulated in 7 out of 8 PCOS samples. CONCLUSION(S): The present data suggest that gene expression profiling may become a useful tool to classify PCOS patients into subgroups with different etiologies. Genome-wide expression profiling using microarrays should be performed to better understand the metabolic derangement(s) in PCOS. PMID- 15820805 TI - Power Doppler ultrasound assessment of ovarian perifollicular blood flow in women with polycystic ovaries and normal ovaries during in vitro fertilization treatment. AB - OBJECTIVE: To evaluate whether ovarian perifollicular blood flow (PFBF) varies by ultrasound among women with polycystic and normal ovaries undergoing in vitro fertilization (IVF). DESIGN: Prospective observational cohort study of women undergoing IVF treatment. SETTING: Department of reproductive medicine at a university teaching hospital. PATIENT(S): Thirty four women with regular spontaneous ovulatory menstrual cycles undergoing IVF divided into two groups according to findings on a baseline transvaginal ultrasound scan: group 1 consisted of 20 women with ultrasound-evident normal ovaries (USNO group), and group 2 consisted of 14 women with ultrasound-evident polycystic ovaries (USPCO group). INTERVENTION(S): Serial transvaginal power Doppler ultrasound assessments throughout the follicular phase of ovarian stimulation. MAIN OUTCOME MEASURE(S): Ovarian PFBF and ovarian stromal artery pulsatility index. RESULT(S): Women with USPCO had a significantly lower ovarian stromal artery pulsatility index at the time of the first ultrasound assessment before starting the FSH injections compared with USNO women. However, there was no difference in ovarian PFBF between women with USPCO and USNO during the follicular phase of ovarian stimulation for IVF. CONCLUSION(S): There is no difference in ovarian follicular vascularity between women with polycystic and normal ovaries during ovarian stimulation at IVF treatment. PMID- 15820806 TI - Placental transfer of rosiglitazone in the first trimester of human pregnancy. AB - OBJECTIVE: To determine the degree of placental transfer of rosiglitazone in early human pregnancy. DESIGN: Prospective observational study. SETTING: University teaching hospital. PATIENT(S): Thirty-one women undergoing surgical termination of pregnancy between 8 and 12 weeks' gestation. INTERVENTION(S): Each woman was given two doses of rosiglitazone (4 mg) before the procedure. MAIN OUTCOME MEASURE(S): Rosiglitazone concentration in fetal tissue and coelomic and amniotic fluids. RESULT(S): The mean maternal serum rosiglitazone concentration was 110.3 +/- 47.9 ng/mL. Rosiglitazone was detectable in 19 fetal samples (61.3%). The mean fetal tissue concentration was 52.7 +/- 26.3 ng/g. Rosiglitazone was more likely to be detected in fetal tissue if the gestation at termination was 10 weeks or more compared with earlier gestation. Coelomic fluid was obtained in 22 cases, and rosiglitazone was detected in 13 samples (59.1%). The mean concentration was 22.8 +/- 7.0 ng/mL. Rosiglitazone was detectable in only two of the 31 amniotic fluid samples, with concentrations of 10.3 and 12.6 ng/mL. CONCLUSION(S): The risk of placental transfer of rosiglitazone is much higher at or after 10 weeks of gestation. Absence of detectable rosiglitazone in amniotic fluid despite its presence in fetal tissue suggests that fetuses may have the ability to metabolize rosiglitazone, and little parent drug was excreted unchanged in urine. PMID- 15820807 TI - Vascular endothelial growth factor gene polymorphisms and idiopathic recurrent pregnancy loss. AB - OBJECTIVE: To investigate whether four common polymorphisms (-2578C/A, -1154G/A, 634G/C, and 936C/T) of the gene encoding for vascular endothelial growth factor (VEGF) are associated with idiopathic recurrent miscarriage. DESIGN: Prospective case-control study. SETTING: University teaching hospital. PATIENT(S): Fifty-two patients with a history of three or more unexplained consecutive pregnancy losses and 82 healthy, postmenopausal controls with at least two live births and no history of pregnancy loss. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Polymerase chain reaction and restriction fragment length polymorphism analysis were performed to identify the different VEGF alleles. RESULT(S): There was a significant difference in the -1154G/A genotype and allele frequency between women with recurrent pregnancy loss and controls. The risk of recurrent pregnancy loss was lower in the carriers of the G allele than in women carrying the A allele (odds ratio = 1.91, 95% confidence interval, 0.12-3.28). No significant association between recurrent spontaneous abortions and -2578C/A, -634G/C, and 936C/T genotypes 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 VEGF gene polymorphisms in women with recurrent miscarriage, demonstrating that the 1154G/A VEGF gene polymorphism is associated with idiopathic recurrent abortions. PMID- 15820808 TI - A mathematical model for evaluation of maternal cell contamination in cultured cells from spontaneous abortions: significance for cytogenetic analysis of prenatal selection factors. AB - OBJECTIVE: To develop a mathematical model for more precise estimation of the incidence of chromosomal abnormalities and the sex ratio among spontaneous abortions masked by maternal cell contamination. DESIGN: Retrospective analysis. SETTING: Academic medical center. PATIENT(S): One hundred twelve samples of spontaneous abortion with a "46,XX" karyotype and 97 parents with aborted embryos. INTERVENTION(S): The presence of Y chromosome DNA in native tissues of "46,XX" spontaneous abortions was detected by amelogenin locus analysis. Detection of aneuploidies in noncultured tissues of "46,XX" abortions was performed by microsatellite DNA analysis and confirmed by fluorescence in situ hybridization. MAIN OUTCOME MEASURE(S): Accuracy of cytogenetic evaluation of spontaneous abortions. RESULT(S): Y chromosome DNA was revealed in 16% of the embryos with a "46,XX" karyotype. According to the mathematical model proposed, the frequency of chromosomal abnormalities in a sample of 478 abortions increased from 54.6% to 60.3%, and the sex ratio in embryos with normal karyotype changed from 0.66 to 1.02. The experimental validation of the model has shown that the observed and expected incidences of chromosomal abnormalities in "46,XX" abortions were in good agreement. CONCLUSION(S): Maternal cell contamination clearly affects the incidence of registered chromosomal abnormalities and the sex ratio in spontaneous abortions. Correction for maternal cell contamination should be taken into account before invoking biological explanations of sex ratio bias and might be useful to include in diagnostic reporting. PMID- 15820809 TI - Laparoscopic management of ovarian remnant. AB - OBJECTIVE: To report outcomes of laparoscopic management of patients with ovarian remnant (OR). DESIGN: Retrospective chart review. SETTING: Referral practice and tertiary medical center. PATIENT(S): Sixty-four patients with confirmed OR who underwent laparoscopic treatment between July 1989 and September 2003. INTERVENTION(S): Laparoscopic excision of OR. MAIN OUTCOME MEASURE(S): Technical feasibility and recurrence. RESULT(S): Sixty-nine laparoscopies were performed to remove ovarian remnants, with five patients requiring two laparoscopies. Two cases were converted to laparotomy and one to mini-laparotomy for bowel resection. In 64% (41 out of 64), pelvic mass was diagnosed by imaging (35 by ultrasound, 5 by computerized tomography [CT], and 1 by both). The majority of ovarian remnants were found attached to one or more of the following: ureter, bowel, pelvic sidewall, bladder, rectum, and uterosacral ligament. Intraoperative complications occurred in four cases: three enterotomy and repair; one cystotomy and repair. Twelve minor postoperative complications occurred including urinary tract infection, hematuria, umbilical incision infection, and transient tachycardia. Three major postoperative complications occurred: one umbilical omental hernia, one wound abscess requiring operation, and one vesicovaginal fistula. Adhesions were present in all cases, endometriosis in 55% (35 out of 64), and fibrosis in 30% (19 out of 64). CONCLUSION(S): In experienced hands, laparoscopic treatment of OR results in acceptable outcomes with its associated advantages over laparotomy. PMID- 15820811 TI - Serum leptin and lactational amenorrhea in well-nourished and undernourished lactating women. AB - OBJECTIVE: To ascertain the possible role of leptin in the resumption of postpartum menstruation in lactating women with differing nutritional statuses. DESIGN: Analysis of data and blood samples collected during a previous prospective study. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Undernourished (body mass index [BMI]< or =19 kg/m(2)) and well-nourished (BMI> or =26 kg/m(2)) lactating women who resumed regular menstruation before 24 weeks and at or after 24 weeks postpartum. INTERVENTION(S): Venous blood samples at four-weekly intervals and other clinical data collected until resumption of regular menstruation. MAIN OUTCOME MEASURE(S): Serum leptin concentrations. RESULT(S): Leptin concentrations were significantly higher in the well-nourished than in the undernourished women, irrespective of the time of resumption of menstruation. Time of resumption of menstruation did not significantly affect leptin levels within well-nourished and undernourished groups. Leptin significantly correlated with BMI (r = 0.78). The BMI (r = -0.53), but not leptin, was significantly and negatively correlated with the duration of lactational amenorrhea. CONCLUSION(S): Leptin is unlikely to be a major determinant of early resumption of regular menstruation in well-nourished women. PMID- 15820810 TI - Serum antimullerian hormone levels best reflect the reproductive decline with age in normal women with proven fertility: a longitudinal study. AB - OBJECTIVE: The aim of this study was to assess which of the basal ovarian reserve markers provides the best reflection of the changes occurring in ovarian function over time (i.e., reproductive aging). DESIGN: Prospective longitudinal study. SETTING: Healthy volunteers in an academic research center. PATIENT(S): Eighty one women with normal reproductive performance during the course of their lives were longitudinally assessed. In this select group of women, becoming chronologically older was considered as a proxy variable for becoming older from a reproductive point of view. INTERVENTION(S): The women were assessed twice, with on average a 4-year interval (T(1) and T(2)). The number of antral follicles on ultrasound (AFC) and blood levels of antimullerian hormone (AMH), FSH, inhibin B, and E(2) were assessed. MAIN OUTCOME MEASURE(S): Longitudinal changes of the markers mentioned and the consistency of these parameters over time. RESULT(S): The mean ages at T(1) and T(2) were 39.6 and 43.6 years, respectively. Although AFC was strongly associated with age in a cross-sectional fashion, it did not change over time. The AMH, FSH, and inhibin B levels showed a significant change over time, in contrast to E(2) levels. The AMH and AFC were highly correlated with age both at T(1) and T(2), whereas FSH and inhibin B predominantly changed in women more than 40 years of age. To assess the consistency of these parameters over time, we investigated whether a woman's individual level above or below the mean of her age group at T(1) remained above or below the mean of her age group at T(2). Serum AMH concentrations showed the best consistency, with AFC as second best. The FSH and inhibin B showed only modest consistency, whereas E(2) showed no consistency at all. CONCLUSION(S): These results indicate that serum AMH represents the best endocrine marker to assess the age-related decline of reproductive capacity. PMID- 15820812 TI - Genital duplex Doppler ultrasonography before and after sexual stimulation in women with sexual dysfunction: gray scale, volumetric, and hemodynamic findings. AB - OBJECTIVE: To report gray scale, volumetric, and duplex ultrasound data before and after sexual stimulation in a large population of women with sexual dysfunction. DESIGN: Retrospective review of women with sexual dysfunction. SETTING: An academic center. PATIENT(S): Pre- and postmenopausal women with sexual dysfunction. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): This is an Institutional Review Board-approved retrospective study of 142 women who underwent duplex Doppler ultrasound (DDU) arousal testing for sexual dysfunction. RESULT(S): This study revealed that DDU was able to record morphologic, volumetric (clitoral and corpora spongiosa diameters), and hemodynamic (peak systolic and end-diastolic) changes in women with sexual dysfunction before and after sexual stimulation. Resistive index measurements, which are indicative of veno-occlusive function, did not significantly change before and after arousal in both the clitoris and corpora spongiosa. We also found a correlation between genital diameter and genital end-diastolic velocity value. Gray scale scanning revealed tunical thickening or plaque in several patients who sustained blunt perineal trauma. CONCLUSION(S): The role of DDU in the management of women with sexual dysfunction remains to be determined; however, end-diastolic velocity values suggest that this parameter has an important physiologic implication as a direct determinant of genital engorgement. PMID- 15820813 TI - Secretion and excretion of human chorionic gonadotropin during early pregnancy. AB - OBJECTIVE: To characterize the profiles of human chorionic gonadotropin (hCG) secretion in blood and its subsequent excretion in urine during conceptive cycles that ended in successful pregnancy and in spontaneous abortion. DESIGN: A prospective study. SETTING: University fertility clinic and research laboratories. PATIENT(S): Healthy, spontaneously ovulating women with regular menses, no history of infertility, and either no male partner or an azoospermic partner. INTERVENTION(S): Blood and urine samples were collected daily from 63 spontaneously ovulating women during 167 cycles of artificial insemination (AI) with donor semen; hCG concentrations were measured in blood and urine, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations were measured in blood by immunoassay. MAIN OUTCOME MEASURE(S): Fecundity, the day of ovulation, the day of hCG detection, and the concentration of hCG on the day of detection in blood and urine. RESULT(S): In 62 conceptions detected, 14 resulted in clinical spontaneous abortion (CAB) and 8 resulted in early pregnancy loss (EPL). When successful pregnancies and pregnancy losses were compared, no significant differences existed between the days of hCG appearance in serum or in urine, the concentrations of hCG on the day of detection, or the incremental change in hCG concentration on the day of detection. CONCLUSION(S): These data validate the use of urinary hCG as a biomarker for assessing peri-implantation pregnancy events. PMID- 15820814 TI - Leuprolide acetate treatment with and without coadministration of tibolone in premenopausal women with menstrual cycle-related irritable bowel syndrome. AB - OBJECTIVE: To evaluate the effectiveness of gonadotropin-releasing hormone agonists (GnRH-a) with or without coadministration of tibolone in women with menstrual cycle-related irritable bowel syndrome (IBS). DESIGN: Prospective, randomized, placebo-controlled clinical trial. SETTING: Universities of Catanzaro and Naples. PATIENT(S): One hundred twenty young premenopausal women with menstrual cycle-related IBS (Rome II criteria). INTERVENTION(S): Administration of leuprolide acetate depot (LAD, 11.25 mg IM/3 months) plus tibolone (group A), LAD plus placebo tablets (group B), and injection of a placebo solution plus placebo tablets (group C). MAIN OUTCOME MEASURE(S): Severity of bowel symptoms or signs of IBS and quality of life (QoL), at baseline and after 6 months of treatment. RESULT(S): In all groups, the mean scores for each symptom or sign of IBS and for QoL were significantly improved after treatment. A significant difference was observed between group C and groups A and B. No difference between these last groups was detected in symptoms or signs of IBS. The QoL scores were significantly higher in group A than in group B. CONCLUSION(S): Gondotropin releasing hormone agonist administration is effective in women with menstrual cycle-related IBS. The addition of tibolone does not reduce effectiveness compared with agonist alone and increases QoL. PMID- 15820815 TI - Perimenopausal androgen decline after oophorectomy does not influence sexuality or psychological well-being. AB - OBJECTIVE: To determine whether oophorectomy during the perimenopause, with the associated decline in ovarian androgens, affects sexual function and psychological well-being negatively. DESIGN: Prospective, observational study comparing sexuality and psychological well-being in women after hysterectomy only (HYST) vs. hysterectomy and concomitant oophorectomy (HYST+BSO). SETTING: University hospital and district general hospital. PATIENT(S): Three hundred sixty-two perimenopausal women scheduled for elective hysterectomy on benign indication were recruited and 323 (89%) completed the 1-year follow-up (217 in the HYST group and 106 in the HYST+BSO group). INTERVENTION(S): The patients were evaluated preoperatively and 1 year after surgery. Postoperatively, estrogen replacement therapy was recommended to all women in the HYST+BSO group and to HYST group subjects with climacteric symptoms. MAIN OUTCOME MEASURE(S): Sex steroids (T, androstenedione, DHEA-S, and E(2)) and sex hormone-binding globulin (SHBG) were measured. Free androgen index and free E(2) index were calculated. Sexuality (McCoy's Female Sex Questionnaire) and psychological well-being (Psychological General Well-Being Index) were evaluated. RESULTS(S): Preoperatively, no hormonal differences were found between the two groups. At 1 year follow-up, all sex steroid levels and indices were decreased and SHBG was increased in the HYST+BSO group. Ovarian sex steroids were decreased in the HYST group, whereas DHEA-S and SHBG were unaltered. Sexuality was unaltered in the HYST+BSO group, whereas decreased scores were found in 3 of 14 sexual variables in the HYST group. Psychological well-being was improved in both groups. There were no correlations between the observed changes (data 1 year after surgery, compared with preoperative data) in androgen levels and index and the observed changes in any aspect of sexuality or psychological well-being. CONCLUSION(S): Hormonal changes after oophorectomy in conjunction with perimenopausal hysterectomy do not significantly change postoperative (1-year) sexual or psychological well-being. PMID- 15820816 TI - Fine-needle and large-needle percutaneous aspiration biopsy of testicles in men with nonobstructive azoospermia: safety and diagnostic performance. AB - OBJECTIVE: To assess by ultrasonography the safety of the association fine-needle aspiration (FNA) plus a new percutaneous large-needle aspiration biopsy technique (LNAB), and to compare the results obtained with the two techniques in the same testicle and with FNA in the two testicles of the same patient. DESIGN: Retrospective analysis of clinical, ultrasonographic, and pathologic data. SETTING: Clinical and academic research environment. PATIENT(S): Three hundred and eighty-seven testicles of 233 patients (ages 19 to 57 years) with nonobstructive azoospermia were consecutively examined with FNA (154) or FNA + LNAB (233); 54 patients (89 testicles) examined with FNA + LNAB underwent a second ultrasonography up to 63 days after. INTERVENTION(S): Ultrasonography, FNA, LNAB. MAIN OUTCOME MEASURE(S): Hypoechoic area at ultrasonography; cytologic picture at FNA or LNAB of only Sertoli cells (category 1), spermatogenetic cells different from elongated spermatids or spermatozoa (category 2), and elongated spermatids and/or spermatozoa (category 3). RESULT(S): Among the 54 patients monitored by ultrasonography 48 (88.9%) showed no difference and 6 (11.1%) showed an hypoechoic area < or =1 cm in the second image. Category 1 occurred at FNA or LNAB in a very similar number of testicles (36% vs. 36.9%). Categories 2 and 3 were more frequent at LNAB (29.9% vs. 9.8%) and at FNA (54.2% vs. 33.2%), respectively. One hundred and sixty-six patients had both testicles examined by FNA; 147 pair of testicles were concordant for the presence or the absence of spermatozoa or elongated spermatids. In the remaining 19 patients (11.4%), these spermatogenetic cells were shown only in the specimen from one of the two testicles. These 19 patients accounted for 20.0% of 93 patients with spermatogenetic cells in at least one of the two testicles. CONCLUSION(S): The combination of FNA and LNAB did not produce clinically or subclinically relevant complications. No important differences in the identification of category 1 with FNA or LNAB were found. Fine-needle aspiration was more adequate in identifying category 3, which was particularly relevant in 20% of the men who had these cells in at least one of the two testicles. PMID- 15820817 TI - A multicenter study evaluating the flowcytometric-based kit for semen analysis. AB - OBJECTIVE: To evaluate the precision and accuracy of a flowcytometric-based semen analysis kit and its application in four fertility centers, as compared with routine microscopic evaluation. DESIGN: A prospective comparative study. SETTING: Four fertility centers, located in Israel and the United States. PATIENT(S): Patients referred to fertility clinic for sperm evaluation. INTERVENTION(S): The precision of semen analysis by both methods was evaluated by inter- and intra technician studies. The accuracy of the sperm counts obtained was assessed by counting a diluted sample and comparing the obtained results with the expected results according to the dilution factor. MAIN OUTCOME MEASURE(S): Sperm count, morphology, motility, viability, white cell count, antisperm antibodies. RESULT(S): The flowcytometric-based kit for semen analysis is more precise and accurate than the manual routine method. Smaller coefficient of variance (CV%) with results were obtained by the flowcytometric-based kit for all semen parameters as compared with the routine manual methods. Sperm density values determined by the flowcytometric-based kit correlated better with the dilution factor. The agreement rates between the flowcytometric and manual methods are sperm count 90%, motility 82%, vitality 70%, round cell counts 78%, and sperm bound antibodies 100%. CONCLUSION(S): Semen analysis by flowcytometric-based kit is advantageous because of improved precision and accuracy compared with the routine method. It provides similar clinical information to that obtained by routine method for sperm count, motility, and antisperm antibodies, and more accurate results for round cell counts (labeled all subclasses) and vitality. PMID- 15820818 TI - Fitz-Hugh and Curtis syndrome-like diaphragmatic endometriosis. AB - We report a case of perihepatic adhesion due to severe endometriosis. PMID- 15820819 TI - Endotoxic shock after gamete intrafallopian transfer. AB - OBJECTIVE: To report an extremely rare case of endotoxic shock due to Enterobacter cloacae that occurred after laparoscopy for gamete intrafallopian transfer (GIFT) in a nulligravid woman. DESIGN: Private fertility center in Cape Town, South Africa. SETTING: Case report. PATIENT(S): A 34-year-old woman with primary infertility. INTERVENTION(S): Routine preparation for GIFT procedure, sonar aspiration to obtain oocytes, followed by GIFT. Endotoxic shock developed within hours after the procedure, followed by admission to the intensive care unit, intravenous antibiotic therapy, mechanical ventilation, and abdominal hysterectomy. MAIN OUTCOME MEASURE(S): Preventing patient mortality and morbidity. RESULT(S): Discharge from the intensive care unit occurred on day 11 after GIFT, which was day 8 after surgery. A MEDLINE search (1980 to 2003) found no previous literature on endotoxic shock associated with assisted reproduction. CONCLUSION(S): Gram-negative infection with subsequent endotoxic shock after assisted reproductive techniques is extremely rare. As this case report shows, early diagnosis and active management of these cases are mandatory to prevent serious complications and mortality. PMID- 15820820 TI - True hermaphroditism presenting as bilateral gynecomastia in an adolescent phenotypic male. AB - OBJECTIVE: To report a rare case of true hermaphroditism presenting in adolescence as bilateral gynecomastia in a phenotypic male. DESIGN: Case report. SETTING: Academic medical center. PATIENT(S): A 19-year-old phenotypic male who presented with pronounced bilateral gynecomastia and was ultimately diagnosed with true hermaphroditism. INTERVENTION(S): Endoscopic removal of the internal female genital tract (hemiuterus and ovary), reduction mammoplasty, repair of penile chordee, and left testicular prosthesis placement. MAIN OUTCOME MEASURE(S): Diagnosis and treatment of true hermaphroditism. RESULT(S): Bilateral mammoplasties, repair of a penile chordee, placement of a left testicular prosthesis, and excision of the left hemiuterus and ovary resulted in return to testicular function in this true hermaphrodite. CONCLUSION(S): Although rare, true hermaphroditism should be suspected in male patients presenting with bilateral breast enlargement in adolescence. PMID- 15820821 TI - Normal birth after transfer of cryopreserved human embryos generated by microinjection of cryopreserved testicular spermatozoa into cryopreserved human oocytes. AB - OBJECTIVE: To report the first birth after transfer of cryopreserved embryos generated by intracytoplasmic sperm injection of cryopreserved testicular spermatozoa into cryopreserved human oocytes. DESIGN: Case report. SETTING: Tertiary center for reproductive technology. PATIENT(S): A 36-year-old woman with primary infertility of 3 years' duration and a 37-year-old man with congenital bilateral absence of the vas deferens. INTERVENTION(S): Cryopreservation of human embryos after oocytes and sperm thawing. MAIN OUTCOME MEASURE(S): Live birth. RESULT(S): A healthy, normal female infant with a birth weight of 2,950 g was born by cesarean section at 38 weeks' gestation, with normal 6-month follow-up. CONCLUSION(S): Embryo cryopreservation can lead to successful results, even with the use of cryopreserved gametes. PMID- 15820822 TI - Embryo cryopreservation after diagnosis of stage IIB endometrial cancer and subsequent pregnancy in a gestational carrier. AB - OBJECTIVE: To describe a case of embryo cryopreservation before hysterectomy and bilateral salpingo-oophorectomy for endometrial cancer. DESIGN: Case report. SETTING: University and community service. PATIENT(S): An infertile woman with endometrial biopsy demonstrating grade II/III moderately differentiated endometrial adenocarcinoma. INTERVENTION(S): A Progestasert intrauterine device (IUD) was inserted into the uterine cavity to potentially reduce tumor proliferation during the stimulation cycle followed by oocyte retrieval and cryopreservation of 14 embryos. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Successful pregnancy in a gestational carrier. CONCLUSION(S): Embryo cryopreservation and use of a gestational carrier may offer a fertility option for patients with endometrial malignancies without substantially delaying treatment. PMID- 15820823 TI - Live birth of healthy triplets after in vitro fertilization and embryo transfer in an acromegalic woman with elevated growth hormone. AB - OBJECTIVE: To report a case of successful triplet pregnancy in an acromegaly patient with IVF and embryo transfer. DESIGN: Case report. SETTING: Tertiary center for assisted reproductive technologies. PATIENT(S): A 33-year-old woman with acromegaly and primary infertility. INTERVENTION(S): Controlled ovulation hyperstimulation and IVF/embryo-transfer treatment using gonadotropins. MAIN OUTCOME MEASURE(S): Uncomplicated pregnancy course and successful take-home triplet infant girls. RESULT(S): Successful ovarian stimulation and IVF is possible in acromegalic patients. CONCLUSION(S): There may be an effect of elevated growth hormone resulting in increased ovarian response to gonadotropin, inducing hyperstimulation and high-order multiple pregnancies in these patients. PMID- 15820824 TI - A case of simultaneous tubal-splenic pregnancy after assisted reproductive technology. AB - OBJECTIVE: To present a case of simultaneous splenic and tubal pregnancy following in vitro fertilization. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 37-year-old woman who had undergone in vitro fertilization and embryo transfer for unexplained infertility at another clinic. INTERVENTION(S): Laparoscopy and laparotomy. MAIN OUTCOME MEASURE(S): Serum concentration of human chorionic gonadotropin (hCG) after salpingosplenectomy. RESULT(S): After transfer of three embryos following fertilization by intracytoplasmic sperm injection (ICSI) at another clinic, the patient was referred to our hospital with suspicion of ectopic pregnancy. Because tubal pregnancy was suspected, laparoscopic right salpingectomy was performed. Although villi were detected in the resected fallopian tube, the serum hCG concentration did not decrease after the operation and a new intraabdominal hemorrhage was detected. We then suspected abdominal pregnancy in the epigastric region, and performed magnetic resonance imaging, computed tomography, and ultrasound examinations, which revealed implantation at the inferior pole of the spleen. Splenectomy was performed, with the resulting disappearance of intraabdominal hemorrhage and rapid fall of the serum concentration of hCG. CONCLUSION(S): Assisted reproduction sometimes results in heterotopic pregnancy, but an abdominal pregnancy involving the upper abdominal organs is considered extremely rare. Particularly, splenic pregnancy is usually overlooked and may only be discovered after a sudden intraabdominal hemorrhage. If hemorrhaging is present in the abdominal cavity but pregnancy is not detected within the pelvis, it is advisable to examine patients using imaging techniques to detect any upper abdominal pregnancies. PMID- 15820825 TI - Effect of midfollicular luteinizing hormone levels on ovarian response and pregnancy outcome in patients undergoing in vitro fertilization in a short-term protocol. AB - In a retrospective study, levels of LH in the midfollicular phase had a significant impact on ovarian response and pregnancy outcome. High LH levels were associated with reduced implantation and clinical pregnancy rates. PMID- 15820826 TI - Trophoblast apoptosis is increased in women with evidence of TH1 immunity. AB - Interferon-gamma secretion from peripheral blood mononuclear cells (PBMCs) of women with unexplained recurrent spontaneous abortions was increased by trophoblast stimulation. Apoptosis was significantly increased in trophoblast cells cultured with the supernatant of PBMCs from women with evidence of T(H)1 immunity to trophoblast stimulation. PMID- 15820827 TI - High frequency of Y chromosome microdeletions in idiopathic azoospermic men with high follicle-stimulating hormone levels. AB - Y chromosome microdeletions were detected in 33.3% (6 out of 18) of idiopathic azoospermic patients with high serum follicle-stimulating hormone (FSH) levels in the present study. The results suggest that it may be necessary to detect microdeletions in patients with azoospermia, especially for those with high serum FSH levels, before assisted reproductive technology (ART) is provided to them. PMID- 15820828 TI - Evaluation of a novel vaginal bromocriptine mesylate formulation: a pilot study. AB - Because of the frequent side effects found with oral bromocriptine, we created two formulas of vaginal bromocriptine suppositories to compare with vaginal application of bromocriptine tablets. The formula containing bromocriptine and a releasing agent (Pluronic F127) showed an increased dissolution rate, 39-fold greater than that of the pure drug alone, and subsequently was effective in lowering serum prolactin. PMID- 15820829 TI - Expression and regulation of thrombospondin-1 by human endometrial stromal cells. AB - Thrombospondin-1 (TSP-1) production was modulated by EGF, IFN-gamma, and in vitro decidualization. It is suggested that TSP-1 may contribute to the regulation of neovascularization in the endometrium and gestational tissues. PMID- 15820830 TI - Evidence for a histaminergic system in the human testis. AB - The complete lack of information about mast cells as a source of histamine and potential target cells for histamine in human testes prompted us to investigate these issues in testes of fertile and infertile patients using a combination of laser microdissection, reverse transcription-polymerase chain reaction (RT-PCR), and immunohistochemistry. We show for the first time the expression of the rate limiting enzyme in histamine synthesis-histidine decarboxylase-by human testicular mast cells and the expression of the histamine (H) receptors 1 (H1) and 2 (H2) by germinal, interstitial, and peritubular cells in the testes of fertile and infertile patients. PMID- 15820831 TI - Insulin growth factors may be implicated in human sperm capacitation. AB - This study was designed to determine whether or not insulin-like growth factor (IGF)-1 and its receptor are present in the human male genital tract. The IGF-1 receptor was found in seminal plasma from fertile and infertile men, but no IGF-1 receptor was observed in sperm from patients with a history of more failed fertilization. The presence of the IGF-1 receptor in the plasma membrane of human sperm is proposed as a future tool for male sterility assessment. PMID- 15820833 TI - Viability of sperm without tails? PMID- 15820834 TI - Technical challenges in freeze-thawing of human ovary. PMID- 15820835 TI - Modeling age at menopause. PMID- 15820837 TI - Increased maternal cardiovascular mortality associated with pregnancy in women with Turner syndrome. AB - In women with Turner syndrome, the risk for aortic dissection or rupture during pregnancy may be 2% or higher, and the risk of death during pregnancy is increased as much as 100-fold. PMID- 15820838 TI - Endogenous opiates and the placebo effect: a meta-analytic review. AB - OBJECTIVE: A meta-analysis was performed to investigate the ability of placebo administration to reduce self-report of pain and to examine whether placebo induced pain reduction might have physiological and psychological underpinnings. METHOD: Forty-five effect sizes and 1183 participants from 12 studies were meta analyzed for the effects of placebo and the opioid antagonist, naloxone, on self report of pain. RESULTS: Analyses showed that placebo administration was associated with a decrease in self-report of pain, and a hidden or blind injection of naloxone reversed placebo-induced analgesia. Furthermore, there were significant between-group differences for type of pain (experimental vs. postoperative/clinical) for placebo studies. CONCLUSIONS: The results support the literature illustrating that the belief and expectation of analgesia induces discrete physiological changes, leading to relief from pain, and this response may be mediated by endogenous opioids. The implications of these findings are discussed in terms of the symbolic aspect of health care and mental health providers' words and context, and their potential impact on the course of illness and well-being. PMID- 15820839 TI - Reconsidering the role of personality in placebo effects: dispositional optimism, situational expectations, and the placebo response. AB - OBJECTIVE: Prior investigations have failed to find reliable personality differences in placebo responding. The present study tests the hypothesis that personality and situational variables interact to determine placebo responding. METHODS: Optimists and pessimists were randomly assigned to one of three conditions. In the first condition, the participants were told that they were to ingest a pill that would make them feel unpleasant (deceptive-expectation group). In the second condition, the participants were told that they were to ingest a pill that would make them feel either unpleasant or was an inactive substance (conditional-expectation group). Finally, a third group was told they were to ingest a pill that was inactive (control group). RESULTS: Pessimists were more likely than optimists to follow a negative-placebo expectation when given a deceptive expectation, but not when given a conditional expectation. CONCLUSION: The personality variable optimism-pessimism relates to placebo responding when individuals are given a deceptive but not a conditional expectation. This suggests that personality and situational variables interact to determine placebo responding. PMID- 15820840 TI - Perceived health status and perceived diabetes control: psychological indicators and accuracy. AB - OBJECTIVE: The aim of this study is to assess the association of psychological, as well as physical and sociodemographic, indicators with patients' ratings of personal health status and diabetes control and to investigate the association of mental health and depression with errors in the perception of diabetes control. METHOD: A sociodemographically diverse sample of 623 diabetes patients was recruited from the general medicine clinics of a county health care system and a Veterans Affairs health care system. We examined three types of determinants of patients' health perceptions: physical health indicators (symptoms, comorbid diagnoses, and glycosylated hemoglobin or HbA1c levels), psychological health indicators (general mental health and diabetes-related worry), and sociodemographic factors (age, race, gender, income, and education). RESULTS: After controlling for patient' sociodemographic characteristics, perceived general health was associated with patients' symptom burden and emotional distress (but not with patients' HbA1c levels). Perceived diabetes control additionally was associated with HbA1c and diabetes-related worries. Further analyses showed that both mental health and diagnosed depression were associated with errors in personal appraisals of diabetes control, with depressed patients more often inaccurately assessing their glycemic control as poor (false-positive error) and nondepressed patients more often missing poor HbA1c levels (false negative error). CONCLUSIONS: Findings indicate that patients use a comprehensive model for assessing their general health and that depression may lead to more accurate assessments of poor glucose control. PMID- 15820841 TI - The association of depression and perceptions of interpersonal relationships in patients with diabetes. AB - OBJECTIVE: Using an instrument assessing interpersonal relationships in patients with diabetes, we hypothesized that a change in depression would be associated with a change in patients' perceptions of themselves and others in relationships. METHODS: Instruments assessing attachment, depression, and demographics were administered twice to 367 patients with diabetes in an HMO primary care setting, 10 months apart. We assessed change in capacity to rely on others (model of other) and to feel worthy of attention (model of self) according to depression change categories (unchanged, decreased, and increased depression). RESULTS: The degree to which patients reported being able to rely on others increased with a reduction in depressive symptoms (P = .02). The degree to which patients endorsed a sense that they were not worthy of attention in relationships increased with an increase in depressive symptoms (P = .02). CONCLUSION: A change in depressive symptoms is associated with a change in perception of interpersonal relationships in patients with diabetes. PMID- 15820842 TI - Somatized depression in primary care attenders. AB - OBJECTIVE: The aim of this study is to determine, within the context of primary care, the frequency of the various ways in which depression is presented with respect to somatic symptoms and to compare depressed patients who present their distress somatically with those with psychological complaints. METHOD: In the two phase cross-sectional study, first, we screened 906 consecutive patients, and second, we interviewed in detail 306 selected patients. RESULTS: The prevalence of depression was 16.8% (CI 95%: 13.4-20.2). There were 59 cases with psychological presentation, 45 somatizers and 16 had organic disorders with depressive comorbidity. Somatizers had lower level of education, and somatized depression was less serious and caused less repercussion. Detection, antidepressive treatment and psychiatric care were lower for somatizers than for psychologizers. CONCLUSIONS: Somatization is a frequent way to present depression in primary care. For somatizers, depression is less severe and is associated with less repercussion. Somatization is associated with the under-detection of the underlying psychiatric process. PMID- 15820843 TI - Anxiety and depression following myocardial infarction: gender differences in a 5 year prospective study. AB - OBJECTIVE: The aim of this study was to examine the impact of the first myocardial infarction (MI) and the relative influence of preexisting confounding factors on anxiety and depression in the following 5 years. METHODS: A total of 23,693 participants, 35-79 years of age at baseline, attended two population based prospective studies in 1984-1986 and in 1995-1997. They underwent physical examination and self-reported demographic, lifestyle, psychosocial, and medical health characteristics in both surveys. Outcome measure was the Hospital Anxiety and Depression rating Scale (HADS). RESULTS: Five hundred twelve participants suffered their first MI in the last 5 years before follow-up. Women showed an increased risk for both anxiety and depression in the first 2 years post-MI, followed by a significant symptom reduction. In contrast, the risk for depression in men increased after 2 years post-MI. Anxiety and depression, low educational level, obesity, daily smoking, and physical inactivity pre-MI significantly predicted a poor psychiatric outcome at follow-up. CONCLUSION: Five-year follow up after MI revealed gender-specific outcomes of anxiety and depression not previously described. PMID- 15820844 TI - Detecting and monitoring depression with a two-item questionnaire (PHQ-2). AB - OBJECTIVE: This study evaluates the two-item Patient Health Questionnaire (PHQ-2) as a measure for diagnosing and monitoring depression. METHODS: We assessed construct validity in a cross-sectional sample of 1619 medical outpatients (mean age 43+/-14 years, 64% female) by comparing the PHQ-2 to four longer self-report questionnaires. Criterion validity was established in a subsample of 520 participants with reference to the Structured Clinical Interview for DSM-IV (SCID). Sensitivity to change was investigated in a prospective study of 167 patients who completed the SCID both at baseline and the 1-year follow-up. RESULTS: With reference to the SCID, the PHQ-2 had a sensitivity of 87% and a specificity of 78% for major depressive disorder and a sensitivity of 79% and a specificity of 86% for any depressive disorder. Its diagnostic performance was comparable with that of longer depression scales. PHQ-2 change scores accurately reflected improved, unchanged, and deteriorated depression outcomes. CONCLUSION: The PHQ-2 performed favorably with respect to a standard diagnostic interview, as well as established depression scales and proved sensitive to change. Thus, the PHQ-2 appears promising as a brief multipurpose measure for detecting depression, grading its severity, and monitoring outcomes over time. PMID- 15820845 TI - Biopsychosocial predictors of fatigue in chronic hepatitis C. AB - OBJECTIVE: The objective of this study was to investigate the relationship of biopsychosocial variables to fatigue in patients with chronic hepatitis C (CHC) with the goal of identifying areas of possible intervention. METHOD: Ninety-four patients with CHC not on antiviral therapy completed a demographic questionnaire and measures of fatigue, health-related quality of life, and depression. Fibrosis stage and relevant medical information were obtained from medical records. RESULTS: A stepwise multiple regression analysis revealed that the most significant predictor of fatigue was poor social functioning, followed by poor physical functioning, greater depression, and female gender. Together, these variables accounted for 68% of the total variance in fatigue. DISCUSSION: Study findings suggest that treatment strategies for fatigue in patients with CHC should focus on increasing social functioning in addition to improving physical functioning and decreasing depressive symptoms. Research investigating the effectiveness of interventions targeting social functioning, such as support groups and supportive counseling, is recommended. PMID- 15820846 TI - Toward an understanding of different manifestations of fatigue. AB - BACKGROUND: The aim of this study was to determine if patient narratives and interviewer reactions are specific in groups of somatic (SF) and psychogenic fatigue (PF) patients. METHODS: The interview criteria and criteria of the interviewer reaction are tested against group classification. Over a 2-year period, 51 (22 PF and 29 SF) patients fulfilling the inclusion criteria were interviewed by two trained independent interviewers. The tape-recorded interviews were analyzed and blind rated by a set of independent physicians. State and Trait Anxiety Inventories (STAI and TRAI, respectively), Beck Depression Inventory (BDI), and Beck's hopelessness scales were completed by the patients. RESULTS: SF versus PF patients more often showed a clear versus vague qualitative description (P=.03), clear changes in levels of intensity (P=.03), and clear factors contributing to the increase (P=.02) and decrease (P=.03) of fatigue. In SF, the raters more often felt calm (P=.03), interested (P=.01), and attentive (P=.02). With PF tapes, the raters more often felt impatient (P=.03), surprised (P=.03), and helpless (P=.01). For the TRAI, the mean score of the PF group was significantly higher compared with that of the SF group (P=.004). For all items, Fisher's Exact Test was used. CONCLUSIONS: Our results confirm significant differences in patient narratives and interviewer reactions for the groups examined. The systematic use of interview criteria and analysis of the interviewer reactions may contribute to the better understanding of fatigue. PMID- 15820847 TI - Relationship between trait-anger and sleep disturbances in middle-aged men and women. AB - OBJECTIVE: The present study was performed to determine whether high levels of trait-anger (T-Anger) are independently associated with sleep disturbances and quantitative and qualitative measurements of sleep in middle-aged adults. METHODS: Enrolled in the study were 2404 men and 2291 women derived from the Korean Health and Genome Study (KHGS), which is an ongoing population-based prospective study. The Spielberger Trait Anger Scale was used to measure T-Anger. RESULTS: Sleep disturbances, including difficulty in initiating and maintaining sleep (DIS and DMS, respectively) and early morning awakening (EMA), significantly increased with increasing levels of T-Anger (P<.05 by test for trend). Nonrestorative feelings in the morning and excessive daytime sleepiness were also strongly associated with higher levels of T-Anger. After adjusting for other potential risk factors, the high and/or moderate T-Anger showed 40% to 70% increases in the odds for sleep disturbances. CONCLUSION: The present study provides evidence that high levels of T-Anger are independently associated with symptoms of disturbed sleep in middle-aged adult population. PMID- 15820848 TI - Coping with body-image threats and challenges: validation of the Body Image Coping Strategies Inventory. AB - OBJECTIVE: Despite extensive research on both body image and coping, little attention has been given to their interface. This investigation examined the reliability and validity of the newly developed Body Image Coping Strategies Inventory (BICSI), which measures how individuals characteristically manage threats or challenges to body-image experiences. METHODS: The BICSI and other relevant body-image and psychosocial functioning inventories were completed by 603 male and female college students. RESULTS: Results revealed a 29-item measure consisting of three internally consistent body-image coping factors: avoidance, appearance fixing, and positive rational acceptance. The BICSI significantly converged with other pertinent measures of body-image evaluation, affect, and investment, and with psychosocial functioning (i.e., self-esteem, social support, and eating disturbance). Regression analyses indicated that multiple coping strategies predicted individuals' body-image quality of life and their eating attitudes. Compared with men, women used all coping strategies more, especially appearance-fixing strategies. Ethnic differences were identified. CONCLUSION: The empirical findings support the reliability and validity of the BICSI. Limitations, implications, and research directions are considered. PMID- 15820849 TI - Preoperative coping strategies and distress predict postoperative pain and morphine consumption in women undergoing abdominal gynecologic surgery. AB - OBJECTIVES: The aim of the present study was to predict postoperative pain and morphine consumption based on preoperative psychosocial factors. METHODS: One hundred and twenty-two women completed measures of distress and coping 1 week before major abdominal gynecological surgery by laparotomy. Forty-eight hours after surgery, measures of pain and negative affect (NA) were completed, and morphine consumption was recorded from a patient-controlled analgesia pump. Four weeks after surgery, measures of pain and NA were completed. RESULTS: Multivariate analyses revealed that preoperative self-distraction coping (P=.039) positively predicted postoperative pain levels in the hospital, after accounting for the effects of age, concurrent NA, and morphine consumption. Emotional support (P=.031) and religious-based coping (P=.036) positively predicted morphine consumption in the hospital, after accounting for the effects of age, concurrent NA, and pain levels. Preoperative distress (P<.04 to .008) and behavioral disengagement (P=.034), emotional support (P=.049), and religious based coping (P=.001) positively predicted pain levels 4 weeks after surgery, after accounting for the effects of age and concurrent NA. CONCLUSION: The results suggest that preoperative psychosocial factors are associated with postoperative pain and morphine consumption. PMID- 15820850 TI - Re: Rief W, Sharpe M. Somatoform disorders. New approaches to classification, conceptualization, and treatment (Editorial). J Psychosom Res 2004; 56: 387-390. PMID- 15820853 TI - Fragile X mental retardation protein levels increase following complex environment exposure in rat brain regions undergoing active synaptogenesis. AB - Fragile X mental retardation protein (FMRP), which is absent in fragile X syndrome, is synthesized in vitro in response to neurotransmitter activation. Humans and mice lacking FMRP exhibit abnormal dendritic spine development, suggesting that this protein plays an important role in synaptic plasticity. Previously, our laboratory demonstrated increased FMRP immunoreactivity in visual cortex of rats exposed to complex environments (EC) and in motor cortex of rats trained on motor-skill tasks compared with animals reared individually in standard laboratory housing (IC). Here, we use immunohistochemistry to extend those findings by investigating FMRP levels in visual cortex and hippocampal dentate gyrus of animals exposed to EC or IC. Rats exposed to EC for 20 days exhibited increased FMRP immunoreactivity in visual cortex compared with animals housed in standard laboratory caging. In the dentate gyrus, animals exposed to EC for 20 days had higher FMRP levels than animals exposed to EC for 5 or 10 days. In light of possible antibody crossreactivity with closely related proteins FXR1P and FXR2P, FMRP immunoreactivity in the posterior-dorsal one-third of cerebral cortex was also examined by Western blotting following 20 days of EC exposure. FMRP levels were greater in EC animals, whereas levels of FXR1P and FXR2P were unaffected by experience. These results provide further evidence for behaviorally induced alteration of FMRP expression in contrast to its homologues, extend previous findings suggesting regulation of its expression by synaptic activity, and support the theories associating FMRP expression with alteration of synaptic structure both in development and later in the life-cycle. PMID- 15820854 TI - Single, but not multiple pairings of sucrose and corticosterone enhance memory for sucrose drinking and amplify remote reward relativity effects. AB - This study tested whether pre-training pairings of ingestion of a 32% sucrose solution and injection(s) of corticosterone (B) would enhance later ingestion in the absence of B, and whether these effects would carry over into later contrast like effects when animals were subsequently shifted to 4% sucrose. Frequency dependence of these pairings was also examined. Three groups of male Sprague Dawley rats were adrenalectomized (ADX). A fourth group was sham ADX. Each ADX group received three presentations of sucrose and B (666 microg/kg, s.c.). One received unpaired presentations (separated by days), one received two unpaired presentations and one paired (i.e., simultaneous) presentation, and one received three paired presentations. Shams received three sucrose presentations paired with saline. Single, but not multiple pairings of B with ingestion of a 32% sucrose solution enhanced later sucrose ingestion, a memorial-like effect that carried over into later, opposite contrast-like effects upon presentation of a less-preferred 4% sucrose solution. These effects could not be easily ascribed to differences in training, other than the pairing regimen itself, nor to motivational differences at the time of testing, and were presumed to be memorial. The pairing and frequency-dependence of these appetitive phenomena are analogous to what is frequently observed during acute or chronic exposure to aversive situations and/or neuromodulatory stress hormones, in terms of their bidirectional effects on memory. Through effects on memory, stress hormones may modulate reward and reward relativity. PMID- 15820855 TI - Modeling behavioral recovery following lesion induction in the rat dentate gyrus. AB - Unilateral entorhinal lesions have enjoyed immense popularity as a model of recovery from damage. In part, the popularity has been supported the laminar organization of the hippocampal formation, which allows for the dissection of the contribution of individual afferent pathways to the recovery process. The commissural/associational pathway is of particular interest, since electrophysiological and gross anatomical data, although limited, have correlated sprouting in this pathway with behavioral recovery. Unfortunately, information relating recovery to synaptic structure is lacking. Addressing this issue, two analyses were conducted. Initially, a quantitative review of the literature reporting behavioral recovery following this type of lesion was conducted using meta-analytic techniques. Using this detailed information across decades of research, multiple linear regression analysis was conducted to address whether the morphological correlates of recovery could predict behavioral recovery. This resulted in an equation relating morphology and recovery that stood up well to several diagnostic tests. Moreover, this model suggests that synapse structure (in particular, synapse size and curvature, as well as terminal compartmentalization and the density of multi-synaptic terminals) holds a greater potential to predict behavioral recovery than increases in synapse number, which is typically seen as the optimal anatomical measure of recovery. This initial attempt to identify, quantify, and validate a model of lesion recovery is an important initial step in understanding how synaptic morphology may help mediate recovery of function. PMID- 15820856 TI - Different types of environmental enrichment have discrepant effects on spatial memory and synaptophysin levels in female mice. AB - Environmental enrichment paradigms that incorporate cognitive stimulation, exercise, and motor learning benefit memory and synaptic plasticity across the rodent lifespan. However, the contribution each individual element of the enriched environment makes to enhancing memory and synaptic plasticity has yet to be delineated. Therefore, the current study tested the effects of three of these elements on memory and synaptic protein levels. Young female C57BL/6 mice were given 3h of daily exposure to either rodent toys (cognitive stimulation) or running wheels (exercise), or daily acrobatic training for 6 weeks prior to and throughout behavioral testing. Controls were group housed, but did not receive enrichment. Spatial working and reference memory were tested in a water-escape motivated radial arm maze. Levels of the presynaptic protein synaptophysin were then measured in frontoparietal cortex, hippocampus, striatum, and cerebellum. Exercise, but not cognitive stimulation or acrobat training, improved spatial working memory relative to controls, despite the fact that both exercise and cognitive stimulation increased synaptophysin levels in the neocortex and hippocampus. These data suggest that exercise alone is sufficient to improve working memory, and that enrichment-induced increases in synaptophysin levels may not be sufficient to improve working memory in young females. Spatial reference memory was unaffected by enrichment. Acrobat training had no effect on memory or synaptophysin levels, suggesting a minimal contribution of motor learning to the mnemonic and neuronal benefits of enrichment. These results provide the first evidence that different elements of the enriched environment have markedly distinct effects on spatial memory and synaptic alterations. PMID- 15820857 TI - Activation of adenosine receptors in the posterior cingulate cortex impairs memory retrieval in the rat. AB - Adenosine A1 and A2A receptor agonists and antagonists have been reported to alter learning and memory. The aim of our study was to investigate the involvement of adenosinergic system in memory retrieval into posterior cingulate cortex (PCC) of Wistar rats. To clarify this question, we tested specifics agonist and antagonists of adenosine A1 and A2A receptors in rats submitted to a one-trial inhibitory avoidance task. The stimulation of adenosine A1 and A2A receptors by CPA and CGS21680, respectively, impaired memory retrieval for inhibitory avoidance task, into PCC. These findings provide behavioral evidence for the role of adenosinergic system in the memory retrieval into PCC. PMID- 15820858 TI - Effects of multiple exposures to D-cycloserine on extinction of conditioned fear in rats. AB - Previous research has shown that an acute, post-training injection of D cycloserine (DCS) facilitates extinction of conditioned fear in rats; however, the effects of multiple exposures to DCS in this situation are not known. In Experiment 1, rats were conditioned (light-shock pairings) and 24 h later given six extinction (light-alone) trials followed by an injection of DCS (15 mg/kg) or saline. The next day, all rats were tested for light-elicited freezing. In Experiment 2, the effect of DCS on extinction was tested in the same manner, except that rats were pre-exposed to DCS (0, 1, or 5 injections) just prior to conditioning. In Experiment 3, rats received five pre-exposures of DCS but conditioning occurred either 2 or 28 days after the last pre-exposure. The results showed that DCS facilitated extinction of conditioned freezing to the light CS when no drug pre-exposure had occurred, but pre-exposure to DCS just prior to conditioning disrupted the facilitation of extinction effect. When 28 days were interposed between pre-exposure and conditioning, the facilitatory effects of DCS on extinction were restored. These findings suggest that DCS has significant clinical value but that behavioral desensitization may occur with multiple exposures; however, desensitization is not permanent and is reduced by the passage of time. PMID- 15820859 TI - Differential activity profile of cAMP-dependent protein kinase isoforms during long-term memory consolidation in the crab Chasmagnathus. AB - The isoforms of cAMP-dependent protein kinase (PKA) show distinct biochemical properties and subcellular localization, suggesting different physiological functions, and conferring the fine-tuning between the activation of cAMP-PKA cascade and the cellular response. The critical role of PKA in memory and synaptic plasticity has been extensively demonstrated both in vertebrates and invertebrates, but the role of PKA isoforms is a matter of debate. Here we present experimental data showing differential PKA activation profiles after two different experiences: an instance of associative contextual learning (context signal learning) and a single exposure to a novel context, both in the learning and memory model of the crab Chasmagnathus. Differences were found in the temporal course of activation and in the involvement of PKA isoforms. We found increased PKA activity immediately and 6 h after context-signal training correlating with the critical periods during which pharmacological inhibition of PKA disrupts memory formation. In contrast, PKA activity increased immediately but not 6 h after single exposure to a novel context. The amounts of PKA I and PKA II holoenzymes were analyzed to determine changes in holoenzyme levels and/or differential activation induced by both experiences. Results indicate that context-induced PKA activation is at least in part due to PKA II, and that PKA activation 6 h after context-signal learning coincides with an increase in the total level of PKA I. Considering the higher sensitivity of PKA I to cAMP, its increment can account for the PKA activation found 6 h after training and is proposed as a novel mechanism providing the prolonged PKA activation during memory consolidation. PMID- 15820860 TI - Inhibition of endogenous carbon monoxide production induces transient retention losses in the day-old chick when trained using a single trial passive avoidance learning task. AB - Carbon monoxide (CO) is most often thought of as an exogenous toxin rather than as a possible endogenous nootrope. However, a limited number of studies have suggested that CO is necessary in memory processing for at least some tasks. While nitric oxide (NO) and CO are known activators of guanylyl cyclase (GC), only the effect of NO on GC has been extensively investigated as a mechanism underlying memory processing. The aim of the present study was to determine if inhibition of CO production would have an effect on memory processing. Using chicks trained on a single trial passive avoidance task, inhibition of CO production using zinc (II) deuteroporphyrin IX 2,4-bis ethylene glycol (ZnBG; 5 microM) resulted in two transient retention losses occurring at around 40 and 130 min post-training. The timing of these transient retention losses was similar to those observed following inhibition of GC, using the same species and task in a previous study. This supports the notion that CO is necessary in memory processing for this task and may act through a GC-dependent mechanism. As ZnBG also directly inhibits GC or nitric oxide synthase (NOS) at high concentrations, a second experiment was carried-out to confirm the specificity of ZnBG for heme oxygenase (HO) at the concentration used. The action of ZnBG was challenged with the HO agonist hemin (100 microM) and the transient deficits were abolished. This confirmed that the action of ZnBG on memory was through a CO-related mechanism rather than directly on GC or NOS. In this way the specificity of ZnBG (5 microM) for HO could be confirmed. The results support a role for endogenous CO in memory processing, possibly through activation of GC. In addition, the transient retention losses observed following administration of ZnBG suggest that CO may be necessary for memory retrieval and not formation as previously thought. PMID- 15820861 TI - Aging affects both perceptual and lexical/semantic components of word stem priming: An event-related fMRI study. AB - In this event-related fMRI study, brain activity patterns were compared in extensive groups of young (N=25) and older (N=38) adults, while they were performing a word stem completion priming task. Based on behavioral findings, we tested the hypothesis that aging affects only the lexical/semantic, but not the perceptual component of word stem priming. To this end, we distinguished between priming-related activity reductions in posterior regions involved in visual processing, and regions associated with lexical/semantic retrieval processes, i.e., left lateral temporal and left prefrontal regions. Both groups revealed significant priming-related response time reductions. However, in accordance with earlier findings, a larger priming effect was found in the group of young participants. In line with previous imaging studies, the groups showed common priming-related activity reductions in the anterior cingulate, and the left inferior prefrontal cortex extending into the anterior portion of the left superior temporal gyrus, and at lower thresholds also in the right occipital lobe. However, when directly comparing the groups, greater priming-related reductions were found for the young group in the left anterior superior temporal gyrus and the right posterior occipital lobe. These findings suggest that, converse to current psychological views, aging affects both perceptual and lexical/semantic components of repetition priming. PMID- 15820862 TI - How hydrides misled chemists. AB - Hydrogen-containing molecules are simple enough to be attractive subjects in experimental diffraction and spectroscopic studies and in quantum computations. Yet, the inferences about molecular structure and force fields originally drawn from studies of these subjects were significantly flawed. In recent developments the original models of structure invoked, such as hybridization, have been superseded. The reasons for this are briefly reviewed. What has emerged to account for molecular geometry, prevailing even over the popular VSEPR theory, is a model of geminal nonbonded interactions. PMID- 15820863 TI - On the vibrational spectra and structural parameters of methyl, silyl, and germyl azide from theoretical predictions and experimental data. AB - The infrared and Raman spectra of methyl, silyl, and germyl azide (XN3 where X=CH3, SiH3 and GeH3) have been predicted from ab initio calculations with full electron correlation by second order perturbation theory (MP2) and hybrid density function theory (DFT) by the B3LYP method with a variety of basis sets. These predicted data are compared to previously reported experimental data and complete vibrational assignments are provided for all three molecules. It is shown that several of the assignments recently proposed [J. Mol. Struct. (Theochem.) 434 (1998) 1] for methyl azide are not correct. Structural parameters for CH3N3 and GeH3N3 have been obtained by combining the previously reported microwave rotational constants with the ab initio MP2/6-311+G(d,p) predicted values. These "adjusted r0" parameters have very small uncertainties of +/-0.003 A for the XH distances and a maximum of +/-0.005 A for the heavy atom distances and +/-0.5 degrees for the angles. The predicted distance for the terminal NN bond which is nearly a triple bond is much better predicted by the B3LYP calculations, whereas the fundamental frequencies are better predicted by the scaled ab initio calculations. The results are discussed and compared to those obtained for some similar molecules. PMID- 15820864 TI - 1,2-Dibromoethyl-trichlorosilane (CH2BrCHBrSiCl3): conformational structure and vibrational properties by gas-phase electron diffraction, infrared and Raman spectroscopy, and ab initio molecular orbital and density functional theory calculations. AB - The molecular structure and conformational properties of 1,2-dibromoethyl trichlorosilane (CH2BrCHBrSiCl3) have been investigated using gas-phase electron diffraction (GED) data recorded at a temperature of 100 degrees C, together with ab initio molecular orbital (MO) and density functional theory (DFT) calculations, infrared (IR) and Raman spectroscopy in the liquid and solid phases, and normal coordinate analysis (NCA). The molecule exists in the gas- and liquid phases as a mixture of three conformers, gauche(-) [G(-)], with a refined torsion angle phi(BrCCBr)=-71(6) degrees, anti [A], with a torsion angle phi(BrCCBr) approximately -170 degrees , and gauche(+) [G(+)], with a torsion angle phi(BrCCBr) approximately +70 degrees . The second torsion angle of importance, the rotation about the CSi bond, has been refined to a value of +175(13) degrees . Torsion angles were only refined for the more abundant G(-) conformer. In the solid phase, only the G(-) conformer was observed. The temperature-dependent Raman spectra have provided an estimate of the relative conformational entropies, DeltaS. The obtained composition from GED refinements was (%) G(-)/A/G(+)=64(27)/23(13)/13(18) (values with estimated 2sigma uncertainties), giving a conformational stability order in agreement with both the Raman enthalpy measurements and the ab initio MO and DFT calculations using the 6-311G(d) basis set and scaled zero-point energies. Relevant structural parameter values obtained from the GED refinements (with the ab initio HF values used as constraints) were as follows (G(-) values with estimated 2sigma uncertainties): bond lengths (r(g)):r(C-C)=1.501(18)A, r(SiC)=1.865(15)A, r(CBr)=1.965(8)A (average), r(SiCl)=2.028(3)A (average). Bond angles ( anglealpha):angleCCSi=114.1(33) degrees , angleC1C2Br=114.0(21) degrees , angleCSiCl=109.6(7) degrees (average). Experimental IR/Raman and obtained vibrational wavenumbers based on both the unscaled, fixed-scaled as well as the scale-refined quantum-mechanical force fields [HF/6-311G(d)] are presented. The results are discussed and compared with some similar molecules from the literature. PMID- 15820865 TI - Conformation of some biologically active aromatic ureas. AB - Experimental IR spectroscopic data for the N-H stretching mode frequencies for several types of tri-substituted ureas containing benzyl and/or phenyl substituents as well as theoretical results from B3LYP/6-31G(d,p) computations on selected compounds provide sufficient evidence to determine the conformational state of these molecules. Two types of N-H bands may be found the spectra: (a) A type band due to a classical trans conformation (trans I) of the CONH structure; (b) B type band arising from an alternative trans form (trans II), in which the N H band is involved in a hydrogen bond like interaction with the aromatic ring at the neighbouring nitrogen atom (benzyl or phenyl substituents). The N-H band of trans ICONH structure is observed at frequencies higher than 3460 cm-1, the actual position depending on weather the non-substituted N-H group is linked to aryl or alkyl substituents. The N-H band of the trans II rotameric structure is observed at 3430-3420 cm-1. PMID- 15820866 TI - Determination of the absolute configuration of chiral alpha-aryloxypropanoic acids using vibrational circular dichroism studies: 2-(2-chlorophenoxy) propanoic acid and 2-(3-chlorophenoxy) propanoic acid. AB - The enantiomers of 2-(2-chlorophenoxy) propanoic acid and 2-(3-chlorophenoxy) propanoic acid were resolved on a chiral HPLC column and investigated using mid infrared vibrational circular dichroism (VCD). Experimental infrared vibrational absorption and VCD spectra were measured in CDCl3 solution in the 2000-900 cm-1 region and compared with the ab initio predictions of absorption and VCD spectra. The predicted spectra were obtained with density functional theory using B3LYP/6 31G* basis set for the stable and dominant conformers. But the predicted spectra did not provide unambiguous structural information due to intermolecular hydrogen bonding in solution. To eliminate the hydrogen bonding effects, the acids were converted to the corresponding methyl esters and the experimental absorbance and VCD spectra of methyl esters were measured. B3LYP predicted spectra were also obtained for the stable and dominant conformers of the esters. From a comparison of the experimental VCD spectra of methyl esters with corresponding ab initio predictions, the absolute configurations of esters, and therefore of their parent acids, are unambiguously determined to be (+)-(R). PMID- 15820867 TI - Vibrational spectroscopic studies, conformations and ab initio calculations of 3,3,3-trifluoropropyltrichlorosilane. AB - Infrared spectra of 3,3,3-trifluoropropyltrichlorosilane (CF3CH2CH2SiCl3) were obtained in the vapour, amorphous and crystalline solid phases in the range 4000 50 cm-1. Additional spectra in argon matrices at 5.0 K were recorded before and after annealing to 20-36 K. Raman spectra of the compound as a liquid were recorded at various temperatures between 298 and 210 K and spectra of the amorphous and crystalline solids were obtained. The spectra suggested the existence of two conformers (anti and gauche) in the fluid phases and in the matrix. When the vapour was shock-frozen on a cold finger at 80 K and subsequently annealed to 120-150 K, six weak or very weak Raman bands vanished in the crystal. Similar variations were observed in the corresponding infrared spectra after annealing and four very weak IR bands disappeared after crystallization. From intensity variations between 298 and 210 K of three Raman band pairs an average value Delta(conf)H degrees (gauche-anti)=6.1+/-0.5 kJmol-1 was obtained in the liquid. Annealing experiments indicate that the anti conformer also has a lower energy in the argon matrices. The conformational equilibrium is highly shifted towards anti in the liquid, and the low energy conformer also forms the crystal. The spectra of the abundant anti conformer and the few bands ascribed to the gauche conformer have been interpreted. Ab initio calculations at the HF/6-311G(**) and B3LYP/6-311G(**) gave optimized geometries, infrared and Raman intensities and vibrational frequencies for the anti and gauche conformers. The conformational energy differences derived were 11.8 and 9.2 kJmol-1 from the HF and the B3LYP calculations, respectively. PMID- 15820868 TI - Matrix-isolation FT-IR spectroscopic study and theoretical DFT(B3LYP)/6-31++G** calculations of the vibrational and conformational properties of tyrosine. AB - The complicated conformational isomerism of tyrosine is studied by experimental matrix-isolation FT-IR spectroscopy combined with theoretical DFT(B3LYP)/6 31++G** calculations. Not less than 18 possible conformations of tyrosine have been considered theoretically. The results revealed that the most and the less stable forms of neutral tyrosine have the same conformation of the main part of amino acid (conformation II) but they differ in orientation of the phenyl ring. The calculated values of the relative energies suggest that all conformations would be detectable in the experimental spectrum. However, it appeared that it is not possible to distinguish in the experimental spectrum between the bands due to the forms with the same conformation of the main part of amino acid but a different orientation of the phenol ring. PMID- 15820869 TI - Is 2-cyclopropylpropene really gauche? AB - Infrared spectra of gaseous and solid 2-cyclopropylpropene (2-CPP, c-C3H5C (CH3)CH2) have been recorded from 3500 to 40 cm-1, and Raman spectra (3200-150 cm 1) of the liquid as well as mid-infrared spectra of 2-CPP in liquid krypton solution (from -105 to -150 degrees C) were also obtained. Ab initio calculations, with basis sets up to 6-311+G(2df, 2pd), were carried out for this molecule, using the restricted Hartree-Fock (RHF) approach, with full electron correlation by the perturbation method to second order (MP2(full)) and density functional theory (DFT) by the B3LYP method. The combination of the experimental and computational results (particularly with the higher basis sets) unequivocally identifies the more stable conformer of 2-CPP as the trans form, with the gauche rotamer higher in energy, but also stable. The cis structure of this compound is not observed experimentally, and is predicted by the computational approaches to be a transition state. By studying the temperature variation of two well-resolved sets of conformational doublets of 2-CPP dissolved in liquid krypton, an average enthalpy difference between conformers of 182+/-18 cm-1 (2.18+/-0.22 kJ mol-1) has been determined, with the trans conformation lower in energy in the fluid states, and the sole conformer present in the polycrystalline solid phase. This enthalpy difference corresponds to an ambient temperature conformational equilibrium in the fluid phases of 2-cyclopropylpropene containing approximately 55+/-2% of the more stable trans rotameric form. A complete vibrational assignment for the trans conformer of 2-CPP is given, and many of the bands of the gauche rotamer have also been assigned. Structural parameters, dipole moments, and rotational constants for this molecule have been calculated at the MP2(full)/6-311+G(d,p) level, and these results--as well as the results from the experimental studies--are compared to similar quantities in related compounds. PMID- 15820870 TI - An ab initio and cryospectroscopic study of the hydrogen chloride and boron trifluoride complexes of cyclopropene. AB - Mid-infrared spectra of cyclopropene (c-C3H4) mixtures with HCl or BF3, dissolved in liquefied argon and in liquefied nitrogen have been examined. Evidence was found for the formation of two different isomers of the 1:1 complexes: a pi-type complex in which the interaction occurs with the pi-bond of cyclopropene, and a sigma-type complex in which the electron acceptors interact with one of the CC single bonds. At lower temperatures, indications for the formation of a 1:2 chain type complex were found. Using spectra recorded between 90 and 124 K in liquid argon, the complexation enthalpies for the pi- and sigma-complexes with HCl were determined to be -8.8(3) and -7.9(3) kJ mol-1, respectively. For the pi-complex with BF3 a value of -7.4(3)kJ mol-1 was found. Structural and spectral information on the 1:1 complexes was obtained from B3LYP/6-311++G(d,p) and MP2(FC)/6-31+G(d) calculations. Using Free Energy Perturbation Monte Carlo simulations to calculate the solvent influences, and statistical thermodynamics to account for zero-point vibrational and thermal contributions, the solution enthalpies were transformed into complexation energies with values of -15.5(6) and -13.8(8) kJ mol-1 for the pi- and sigma-isomers of c-C3H4.HCl, respectively, and of -12.7(8) kJ mol-1 for the pi-type BF3 complex. PMID- 15820871 TI - A matrix isolation study of the photochemically induced reactions of nitrogen dioxide with 1,2-dibromoethene and 1,2-dichloroethene using Fourier transform infrared spectroscopy. AB - Photolyses of matrices of either BrCHCHBr/NO2/Ar or ClCHCHCl/NO2/Ar using quartz filtered radiation (lambda>240 nm) led to the appearance of infrared bands attributable to carbonyl, carbon monoxide, and ketene species; no bands belonging to a precursor complex NO2cdots, three dots, centeredXCHCHX (where X=Br or Cl) were observed upon matrix deposition. The possible reaction pathway is discussed. PMID- 15820872 TI - Non-destructive Raman analyses--polyacetylenes in plants. AB - Ferdinand Bohlmann has described the isolation, the identification and the structure elucidation of acetylene compounds in many plants, and confirmed it by its synthesis. We have recorded the Raman spectra of most of these plants non destructively by FT-Raman spectroscopy using radiation at 1064 nm. We could not observe any interfering fluorescence. We found acetylene compounds in some plants, even distinct compounds with different concentration in various parts of it. The distribution of the different compounds over the plant can be observed and their changes during the ontogenesis can be followed by a FT-Raman mapping technique. Of special help is a library of Raman and IR spectra and the structure of the compounds, synthesized by Bohlmann. Thus, the Raman technique allows analyses in a very short time replacing the usual time-consuming separation procedures and avoiding artefacts during clean-up procedures. PMID- 15820873 TI - Ab initio molecular orbital studies of the vibrational spectra of the van der Waals complexes of boron trifluoride with the noble gases. AB - The molecular structures, interaction energies, charge transfer properties and vibrational spectra of the van der Waals complexes formed between boron trifluoride and the noble gases neon, argon, krypton and xenon have been computed using second and fourth order Moller-Plesset perturbation theory and the Los Alamos National Laboratory LANL2DZ basis set. The complexes are all symmetric tops, with the noble gas atom acting as a sigma electron donor along the C3 axis of the BF3 molecule. The interaction energies are all vanishingly small, and the amount of charge transferred in each case is of the order of 0.01e. The directions of the wavenumber shifts of the symmetric bending (nu2) and antisymmetric stretching (nu3) modes of the BF3 fragment confirm those determined experimentally, and the shifts are shown to correlate well with the polarizability of the noble gas atom and the inverse sixth power of the intermonomer separation. The nu2 mode is substantially more sensitive to complexation than the nu3 vibration. PMID- 15820874 TI - Steric and electronic effects on the conformations of n-butane derivatives with trichlorosilyl, silyl and trichloromethyl groups. AB - The molecular structure and conformation of 1,1,1,4,4,4-hexachloro-1,4 disilabutane in the gas-phase have been determined by electron diffraction and computational methods. The lowest-energy conformation has the trichlorosilyl groups anti to one another. The gauche conformation also has a shallow potential minimum, but lies about 19 kJ mol-1 above the anti form. Calculations on related butane derivatives, in which terminal methyl groups have been replaced by CCl3, SiH3 and SiCl3 groups, reveal that the conformational preferences are primarily caused by steric interactions between the terminal groups, and that it is the presence of chlorine atoms that destabilises gauche conformations. The electronegativity of the chlorine atoms has only small effects, mainly limited to the SiCl bond lengths. PMID- 15820875 TI - Density functional theory and empirical derived force fields for the delocalized polaron form of polyaniline. Application to properties determination of an isolated oligomer using molecular dynamics simulations. AB - Normal mode analyses obtained from quantum chemical calculations at the DFT level of theory have been performed for the repetitive unit of the delocalized bipolaron form of polyaniline (PANI). Empirical molecular mechanics force field parameters were consecutively refined using the SPASIBA software and applied to the molecular dynamics properties of an isolated dodeca oligomer model of the leucoemeraldine form of PANI. It is shown that effects of protonation of the emeraldine base spread over the four rings constituting the repeating unit. Molecular dynamics simulations reveal alternative bendings of the whole chain with a time period of 18-20 ps for the model of PANI under study. PMID- 15820876 TI - A cryosolution infrared and ab initio study of the van der Waals complexes of cyclopentene with hydrogen chloride and boron trifluoride. AB - The formation of weak molecular complexes of cyclopentene with HCl and BF3, dissolved in liquid argon is investigated using infrared spectroscopy. Evidence is found for the formation of 1:1 complexes in which the Lewis acid under study binds to the CC double bond. At higher concentrations of HCl, weak absorption bands due to 1:2 species are also observed. From spectra recorded at different temperatures between 92 and 127 K, the complexation enthalpies for CP.HCl and CP.BF3 are determined to be -9.5(3) and -16.1(9) kJ mol-1, while for CP.(HCl)2 a value of -17.0(6) kJ mol-1 is obtained. For the 1:1 and 1:2 complexes, structural and spectral information is obtained from ab initio calculations at the MP2/6 31+G(d) level. Using free energy perturbation Monte Carlo simulations to calculate the solvation enthalpies and statistical thermodynamics to account for zero-point vibrational and thermal contributions, the complexation energies for CP.HCl and CP.(HCl)2 are estimated from the experimental complexation enthalpies to be -17.4(14) and -34.0(20) kJ mol-1, while the value for CP.BF3 was derived to be -23.4(22) kJ mol-1. The experimental complexation energies are compared with the theoretical values derived from the MP2/6-31+G(d) potential energy surfaces and with single point energies calculated at the MP2/6-311++G(3df,2pd) level. PMID- 15820877 TI - Vibrational analyses of vinylsulfonamide CH2CHSO2NH2. AB - The structure and conformational stability of vinylsulfonamide CH2CHSO2NH2 were investigated by DFT-B3LYP/6-311+G** and ab initio MP2/6-311+G** calculations. From the calculations the molecule was predicted to exist predominantly in the gauche-syn (vinyl group nearly eclipses one of the SO bonds and the NH2 and the SO2 moieties eclipse each other) conformation with the possibility of low abundance of the cis-syn and the gauche-anti forms. The asymmetric potential function for the internal rotation about CS bond was determined for the molecule. The vibrational frequencies were computed at DFT-B3LYP level for the gauche-syn conformer of the molecule and its d2(C2H3SO2ND2) and d3(C2D3SO2NH2) deuterated species. Normal coordinate calculations were then carried out and the potential energy distributions were calculated for the molecule. PMID- 15820878 TI - Raman spectroscopic characterisations and analytical discrimination between caffeine and demethylated analogues of pharmaceutical relevance. AB - The FT Raman spectrum of caffeine was analysed along with that of its demethylated analogues, theobromine and theophylline. The similar but not identical structures of these three compounds allowed a more detailed assignment of the Raman bands. Noticeable differences in the Raman spectra of these compounds were apparent and key marker bands have been identified for the spectroscopic identification of these three compounds. PMID- 15820879 TI - Matrix-isolation FT-IR spectra and theoretical study of dimethyl sulfate. AB - The preferred conformations of dimethyl sulfate and their vibrational spectra were studied by matrix-isolation FT-IR spectroscopy and theoretical methods (DFT and MP2, with basis sets of different sizes, including the quadruple-zeta, aug-cc pVQZ basis). Conformer GG (of C2 symmetry and exhibiting OSOC dihedral angles of 74.3 degrees ) was found to be the most stable conformer in both the gaseous phase and isolated in argon. Upon annealing of the matrix, the less stable observed conformer (GT; with C1 symmetry) quickly converts to the GG conformer, with the resulting species being embedded in a matrix-cage which corresponds to the most stable matrix-site for GG form. The highest energy TT conformer, which was assumed to be the most stable conformer in previous studies, is predicted by the calculations to have a relative energy of ca. 10 kJ mol-1 and was not observed in the spectra of the matrix-isolated compound. PMID- 15820880 TI - Ab initio study of the barriers to methyl torsion and torsional frequencies of acetyl molecules. AB - A wide range of ab initio and hybrid density functional methods and basis sets have been employed to calculate the barriers to methyl internal rotation in a range of molecules with the acetyl moiety. Comparison is made of the computed torsional frequency with the experimental torsional frequency, nu(obs), for each molecule. With the MP2/6-311+G(3df,2p) combination of method and basis set, the agreement is better than 4 cm-1 for most of the molecules, where nu(obs) or the V3 barrier is well-determined experimentally. PMID- 15820881 TI - Quantum-chemical studies of methyl and silyl torsional frequencies and the structures of disilylsulphide and trisilylphosphine. AB - Torsional frequencies of methyl and silyl groups occurring in a range of molecules have been calculated by HF, B3LYP and MP2 methods using several basis sets. Linear correlations between calculated and observed values are derived and used to predict unobserved or dubious frequencies. The current experimental value for the E torsion in trimethylphosphine is questioned. The relative merits of the B3LYP and MP2 methods are explored. MP2 calculations can show wide variation with respect to basis set. In cases where two or more silyl groups are attached to a common atom, as in disilylsulphide (SiH3)2S, disilylmethane (SiH3)2CH2, trisilylmethane (SiH3)3CH and tetrasilylmethane (SiH3)4C, marked differences occur between B3LYP and MP2 estimates. These may be linked to concomitant differences in conformation or potential barrier restraining internal rotation. In disilylmethane the B3LYP results agree much better with experiment than those from the MP2 method. HF and B3LYP calculations for disilylsulphide and trisilylphosphine give normal C2v and C3v equilibrium structures, respectively, but in MP2 structures the silyl groups are twisted through 6-13 degrees yielding C2 and C3 configurations. It may be possible to distinguish between these structures through the observation of isolated SiH stretching frequencies in the spectra of fully deuterated materials. Dispersion forces could contribute to the twisting calculated by the MP2 method. Further studies of the microwave and vibrational spectra of disilylsulphide and trisilylphosphine isotopomers are needed. PMID- 15820882 TI - Dimerization of the keto tautomer of acetohydroxamic acid-infrared matrix isolation and theoretical study. AB - Dimerization of the keto tautomer of acetohydroxamic acid has been studied using FTIR matrix isolation spectroscopy and DFT(B3LYP)/6-31+G(d,p) calculations. Analysis of CH3CONHOH/Ar matrix spectra indicates formation of two dimers in which two intramolecular CO...HON bonds within two interacting acetohydroxamic acid molecules are retained. A chain dimer I is stabilized by the intermolecular CO...HN hydrogen bond, whereas the cyclic dimer II is stabilized by two intermolecular NH...O(H)N bonds. Twelve vibrations were identified for dimer I and six vibrations for dimer II; the observed frequency shifts show a good agreement with the calculated ones for the structures I and II. Both dimers have comparable binding energies (DeltaE(ZPE)(CP)I, II=-7.02, -6.34 kcal mol-1) being less stable than calculated structures III and IV (DeltaE(ZPE)(CP)III, IV=-9.50, 8.87 kcal mol-1) in which one or two intramolecular hydrogen bonds are disrupted. In the most stable 10-membered cyclic dimer III, two intermolecular CO...HON hydrogen bonds are formed at expense of intramolecular hydrogen bonds of the same type. The formation of the less stable (AHA)2 dimers in the studied matrixes indicates that the formation of (AHA)2 is kinetically and not thermodynamically controlled. PMID- 15820883 TI - Gas-phase on-line generation and infrared spectroscopic investigations of polyphosphazenes, (NPX2)3 where X=F, Cl and Br. AB - Gas-phase infrared spectra of polyphosphazenes (phosphonitrilic halides trimer), (NPX2)3 where X=F, Cl and Br have been recorded. The molecules were generated for the first time by an on-line process using solid (NPCl2)3 as a precursor passed over heated sodium fluoride and potassium bromide at about 550 and 700 degrees C for (NPF2)3 and (NPBr2)3 production, respectively. The products were characterized by the infrared spectra of their vapors. The low-resolution gas phase Fourier transform infrared spectra reported for the first time show strong bands centered at 1295, 1215 and 1200 cm-1, assigned to nu7(E'), in plane PN stretching mode of (NPX2)3, where X=F, Cl and Br, respectively. PMID- 15820884 TI - Vibrational spectroscopic characterization of fluoroquinolones. AB - Quinolones are important gyrase inhibitors. Even though they are used as active agents in many antibiotics, the detailed mechanism of action on a molecular level is so far not known. It is of greatest interest to shed light on this drug-target interaction to provide useful information in the fight against growing resistances and obtain new insights for the development of new powerful drugs. To reach this goal, on a first step it is essential to understand the structural characteristics of the drugs and the effects that are caused by the environment in detail. In this work we report on Raman spectroscopical investigations of a variety of gyrase inhibitors (nalidixic acid, oxolinic acid, cinoxacin, flumequine, norfloxacin, ciprofloxacin, lomefloxacin, ofloxacin, enoxacin, sarafloxacin and moxifloxacin) by means of micro-Raman spectroscopy excited with various excitation wavelengths, both in the off-resonance region (532, 633, 830 and 1064 nm) and in the resonance region (resonance Raman spectroscopy at 244, 257 and 275 nm). Furthermore DFT calculations were performed to assign the vibrational modes, as well as for an identification of intramolecular hydrogen bonding motifs. The effect of small changes in the drug environment was studied by adding successively small amounts of water until physiological low concentrations of the drugs in aqueous solution were obtained. At these low concentrations resonance Raman spectroscopy proved to be a useful and sensitive technique. Supplementary information was obtained from IR and UV/vis spectroscopy. PMID- 15820885 TI - Characterization of kaolinite of the Baten El-Ghoul region/south Jordan by infrared spectroscopy. AB - The infrared spectra (3700-500 cm-1) have been recorded for kaolinite obtained from the Baten El-Ghoul region/south Jordan. The spectra were recorded for raw and dried samples, using KBr disk and nujol methods. The vibrational assignments of the basic fundamental modes were proposed, which were supported by the spectra obtained from the two methods used. The results obtained were compared with other related studies. PMID- 15820886 TI - Raman spectra of deuteriated taurine single crystals. AB - The polarized Raman spectra of partially deuteriated taurine [(ND3+)0.65(NH3+)0.35(CH2)2SO3-] crystals from x(zz)x and x(zy)x scattering geometries of the Ag and Bg irreducible representations of the factor group C2h are reported. The temperature-dependent Raman spectra of partially deuteriated taurine do not reveal any evidence of the structural phase transition undergone by normal taurine at about 250 K, but an anomaly observed in the 180 cm-1 band at approximately 120 K implies a different dynamic for this band (which is involved in a pressure-induced phase transition) in the deuteriated crystal. PMID- 15820887 TI - Near infrared Raman spectra of human brain lipids. AB - Human brain tissue, in particular white matter, contains high lipid content. These brain lipids can be divided into three principal classes: neutral lipids including the steroid cholesterol, phospholipids and sphingolipids. Major lipids in normal human brain tissue are phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, sphingomyelin, galactocerebrosides, gangliosides, sulfatides and cholesterol. Minor lipids are cholesterolester and triacylglycerides. During transformation from normal brain tissue to tumors, composition and concentration of lipids change in a specific way. Therefore, analysis of lipids might be used as a diagnostic parameter to distinguish normal tissue from tumors and to determine the tumor type and tumor grade. Raman spectroscopy has been suggested as an analytical tool to detect these changes even under intra-operative conditions. We recorded Raman spectra of the 12 major and minor brain lipids with 785 nm excitation in order to identify their spectral fingerprints for qualitative and quantitative analyses. PMID- 15820888 TI - Raman study of structural relaxation and boson peak in amorphous films of adamantane. AB - Low-frequency Raman scattering was used to study amorphous solid films of adamantane, a globular non-polar hydrocarbon molecule. As evidenced by its spectral characteristics this type of disorder is different from the orientational disorder found in the room temperature plastic phase by the absence of the translational order as well. This gives rise to the boson peak related to acoustic phonons which gradually disappears upon heating with simultaneous emerging of the phonon line at 50 cm-1 which characterizes the low-temperature ordered phase of adamantane. Adamantane dynamics resembles that of C60 fullerene although not in the same temperature range. All this makes adamantane an attractive system that could serve as a practical reference in molecular simulation studies of the glassy phase of model fluids. PMID- 15820889 TI - Vibrational spectrum, ab initio calculation, conformational equilibria and torsional modes of 1,3-dibromopropane. AB - The infrared and Raman spectrum of 1,3-dibromopropane is reported in the crystalline, liquid and gaseous states. These measurements are compared to the results of ab initio calculations carried out using the 6-31+g* Gaussian basis set for a restricted Hartree-Fock computation. The calculation was repeated using second order Moeller-Ploesset perturbation theory to accommodate electron correlation using the 6-31 g* basis set. The three most stable conformers are GG (C2), AG (C1) and AA (C2v), where A and G stand for anti and gauche orientations of the bromomethyl group relative to the plane of the carbon atoms. The point group symmetry of each structure is given in parentheses. The fourth conformer, G'G (Cs) is of such high energy that it is not observed experimentally in isotropic media in either the infrared or Raman spectrum. In the crystalline state, comparison of the infrared and Raman spectrum with that calculated for the C2 conformer shows that only the GG (C2) conformer survives, and the doublet structure of many of the bands in the spectrum indicates at least two molecules per unit cell. The ab initio calculations predict and the temperature dependence of the Raman spectrum of the liquid confirms that the stability order is C2 or =50% decrease from baseline pain (39% vs 15% for placebo, P = .002). Pregabalin also significantly reduced sleep interference, past week and present pain intensity, sensory and affective pain scores, and bodily pain and decreased by > or =50% the number of patients describing their pain as gnawing, sickening, fearful, and punishing-cruel. More patients receiving pregabalin 600 mg/day than placebo showed improvement, as rated on the Clinical and Patient Global Impression of Change scales, 73% vs 45% and 85% vs 47%, respectively. Pregabalin 150 mg/day was essentially no different from placebo. Dizziness was the most common side effect. These study results show pregabalin 600 mg/day to be safe and effective in reducing the pain and other associated symptoms of painful diabetic neuropathy. PERSPECTIVE: Painful diabetic peripheral neuropathy is a challenging neuropathic pain syndrome. This randomized controlled trial demonstrates that pregabalin, a new drug that interacts with the alpha2-delta protein subunit of the voltage gated calcium channel, is an efficacious and safe treatment for the pain of this condition. PMID- 15820915 TI - Long term hematologic recovery after autologous stem cell transplantation in lymphoma patients: impact of the number of prefreeze and post-thaw CD34+ cells. AB - BACKGROUND: Autologous Stem Cell Transplantation (ASCT) with Peripheral Blood Stem Cells is widely used as consolidation in lymphoma patients. The rapidity and stability of cell engraftment correlate with the number of CD34+ cells in the autograft. However, whether CD34+ cells should be quantified before or after cryopreservation remains unclear. PATIENTS AND METHODS: Of 173 consecutive patients who underwent ASCT in our department from Nov 1, 1995 to Nov 1, 2000, 133 (78 %) were alive without relapse at one year. We report here the results for 106 patients whose hematologic data were available. RESULTS: At one year, the hemoglobin was normal in 47% of the patients, the leukocytes, in 77% and the platelets, in 60%. Only 33% had a normal blood count. We observed a significant correlation between prefreeze and post-thaw CD34+ cell numbers (r = 0.77). However, multivariate analysis using the Cox model with smoothing splines to assess the best cut-off point for these numbers demonstrated that the only independent predictive factor for a normal blood count after one year was a prefreeze number of CD34+ cells above 5.10(6)/kg. CONCLUSION: An optimal long term hematologic recovery after ASCT required a number of prefreeze CD34+ cells of at least 5.10(6)/kg. PMID- 15820916 TI - [Implication of alternative splice transcripts of caspase-3 and survivin in chemoresistance]. AB - Recent studies have shown that resistance to apoptosis may contribute to chemoresistance. Alteration of caspases, such as caspase-3, results on decreased apoptosis. Genes of IAP (inhibitor of apoptosis proteins) family, such as survivin, were also implicated in tumor development where they are mutated or have deregulated expression. Initial studies revealed strong survivin expression in several fetal tissues and some proliferating adult tissues, whereas no survivin expression was detected in a range of adult tissues. Although the factors at the origins on survivin re-expression in tumors are still unknown, the anti-apoptotic function of survivin is mediated in part by inhibiting caspase-3 activity. Recently, functionally divergent splice variants resulting from alternative splicing, with apoptotic (for caspase-3) or anti-apoptotic (for survivin) opposite activities have been described. The alternative splice variant, caspase-3s results from exon 6 deletion and shows antagonist of apoptotic property of caspase-3. Three alternative splice variants of survivin (survivin-DeltaEx3, survivin-2B and survivin-3B) differing in their anti apoptotic properties were reported. While the anti-apoptotic effect of survivin DeltaEx3 is preserved, survivin-2B has lost its anti-apoptotic potential and may act as a naturally occurring antagonist of survivin and survivin-DeltaEx3. At present, little is known about properties of survivin-3B. Several evidences indicate that in several cancers, the ratio of splice variants is significantly altered, and modifications of splicing pathways have been developed for cancer treatment. Recent investigations have shown that expression of alternative splice variants of caspase-3 and of survivin were also altered in many human cancers, and that variations in their expression were associated with tumor progression and chemoresistance. In this article, we describe recent data concerning alternative splice variants of these two proteins. PMID- 15820914 TI - Gonadal hormone modulation of mu, kappa, and delta opioid antinociception in male and female rats. AB - Previous studies suggest that sex differences in morphine antinociception in rodents might be attributed to the activational effects of gonadal hormones. The present study determined whether hormonal modulation of opioid antinociception in adult rats extends to opioids other than the prototypic mu agonist morphine. Male and female rats were sham-gonadectomized (sham-GDX) or gonadectomized (GDX) and replaced with no hormone, estradiol (E2, females), progesterone (P4, females), E2+P4 (females), or testosterone (males). Approximately 28 days later, nociception was evaluated on the 50 degrees C hot plate and warm water tail withdrawal tests before and after subcutaneous administration of hydromorphone, buprenorphine, U50,488, or SNC 80. In sham-GDX (gonadally intact) rats, the mu agonists and U50,488 were less effective in females than in males in at least one nociceptive test, and the delta agonist SNC 80 was less effective in males than in females. In males, gonadectomy tended to decrease, and testosterone tended to increase antinociception produced by 3 of the 4 agonists. In females, gonadectomy and hormone treatment had more variable effects, although E2 tended to decrease mu opioid antinociception. The present results suggest that activational effects of gonadal hormones are relatively modest and somewhat inconsistent on antinociception produced by various opioid agonists in the adult rat. PERSPECTIVE: This study demonstrates that reproductive hormones such as testosterone in males and estradiol in females do not consistently modulate sensitivity to the analgesic effects of opioids in the adult organism. PMID- 15820918 TI - [Metabolism, mechanism of action and resistance to cytotoxic nucleoside analogues]. AB - Cytotoxic nucleoside analogues are widely used in treatment of patients with hematological malignancies as well as for some solid tumors. Resistance developed against these molecules limit their clinical use. Many studies on cell models and clinical samples have identified cellular mechanisms involved in this phenomenon. Here, we describe the available data concerning the proteins involved in the metabolism and the mechanism of action of nucleoside analogues, as well as the clinical studies showing their implication in the resistance to these drugs. PMID- 15820917 TI - [Farnesyltransferase inhibitors: preliminary results in acute myeloid leukemia]. AB - Farnesyltransferase inhibitors (FTIs) are small-molecule inhibitors that selectivly inhibit farnesylation of a number of intracellular substrate proteins such as Ras. Preclinical work has revealed their ability to effectively inhibit tumor growth in vitro and in vivo in animal models across a wide range of malignant phenotypes. Acute myeloid leukemias (AMLs) are appropriate disease targets in that they express relevant biologic targets such as Ras, MEK, AKT, and others that may depend upon farnesyl protein transferase activity to promote cell proliferation and survival. Indeed, different intracellular proteins are substrates for prenylation. Interruption of prenylation may prevent substrates from undergoing maturation which may result in the inhibition of cellular events that depend on the function of those substrates. Phase I trials in AML and myelodysplasia have demonstrated biologic and clinical activities as determined by target enzyme inhibition, low toxicity, and both complete and partial responses. As a result, phase II trials have been initiated in order to further validate clinical activity and to identify downstream signal transduction targets that may be modified by these agents. It is anticipated that these studies will serve to define the optimal roles of FTIs in patients with these hematologic malignancies and provide insight into effective methods by which to combine FTIs with other agents. PMID- 15820919 TI - [News in therapeutic management of chronic lymphoid leukemia]. AB - Chronic lymphocytic leukemia follows an extremely variable clinical course with survival range from months to decades. Some patients present minimal symptoms and others organomegaly requiring rapidly therapy. Therapeutic options take into account efficacy, toxicity and prognostic factors. One of the well known prognosis factor is the clinical staging developed either by Rai et al. or by Binet et al. However, there is an important heterogeneity concerning the course of the disease among patients within a single stage group. Recently, several important observations related to the biologic significance of VH gene mutational status, expression of CD38, over-expression of ZAP-70 and chromosomal aberrations have led to the ability to identify high risk patients with rapid disease progression and lower survival. It has been demonstrated that the VH mutation status is clinically highly relevant. CLL patients with unmutated VH gene show an unfavourable course with a very rapid progression. Specific genomic aberrations have been associated with disease characteristics such as lymphadenopathy related to 11q deletion and resistance to treatment related to 17p deletion. VH gene mutation status and genomic aberrations appear separate parameters when considering their prognostic relevance but nevertheless they are correlated: unfavourable aberrations (11q- and 17p-) occur more frequently in VH unmutated CLL patients. According to these prognostic factors, several treatments including purine analogues and/or monoclonal antibodies have been tested with different schedules and doses of monoclonal antibodies (rituximab and alemtuzumab) considering safety to determine the better efficacy. Infections and haemolytic anemia remain the most frequent complications during conventional chemotherapy. In autologous transplant setting, the transplant related mortality is less than 10%, but survival curve do not show a plateau with about 50% of patients relapsing at 4 years. Conventional allogeneic transplantation could achieve durable remission and probably cure the disease but at the price of a too high transplant related mortality related to depressive immune status and old age of CLL population. In order to minimize the toxicity and to improve graft-versus leukemia effect, development of reduced intensity conditioning (RIC) regimens appear particularly important for CLL patients. Recent studies, although a still short follow-up show very promising results and use of monoclonal antibodies in the conditioning or just after transplant could improve the results of allogeneic stem cell transplantation and cure a larger number of CLL patients. Recent advances to categorize CLL patients according to risk stratification regarding new prognostic factors (FISH, CD38, ZAP70, Ig mutational status) should allow to define better the best therapeutic strategy. In parallel, age, co morbidities and the notion of the risk-adapted strategy have also an important impact adding. PMID- 15820920 TI - [Patients' information in percutaneous core breast biopsy]. AB - Information takes a large part of patient's perceptions of the procedure. If the information is adapted, patients tolerate the procedure well. We point out indications and explain the different types of procedure. For infraclinical masses, sonographically guided automated core needle biopsy removes samples. Sonographically guided vacuum-assisted large-core biopsy is only used for diagnosis generally on second purpose for masses. Stereotaxic vacuum-assisted large-core biopsy is used for microcalcifications (needle 8 or 11 gauge). Haematoma and bleeding are rare (< 4%), but care is necessary to avoid complications. Information is executed before the procedure and a booklet is giving to the women. This information has to be adapted because lot of informations could increase patient's anxiety if they don't want more informations and "not enough information" could also increase anxiety if they want more informations. The practitioner, by hearing, his formation and his experience, is able to adapt informations. PMID- 15820921 TI - [Alternatives to the radiotherapy of stage I testicular seminoma]. AB - Adjuvant irradiation is currently the most frequently used standard treatment for the clinical stage I seminoma (CSI) following orchiectomy. There is a potential carcinogenic risk with irradiation that prompted a search for alternative adjuvant treatment approach. The cure in CSI seminoma patients can be achieved with surveillance or chemotherapy. Surveillance takes into account the fact that 80% of patients do not need any adjuvant treatment after orchiectomy and are overtreated by adjuvant irradiation. Recently, one cycle of adjuvant carboplatin has been proven in a prospective randomized trial. Taken together, all three treatment options are acceptable standard strategies for the management of patients with CSI. Finally, the experience with surveillance strategy allowed an in-depth meta analysis of factors predictive for relapse discrimining the patients who are in need of post orchiectomy adjuvant treatment from those who safety can be followed by the surveillance strategy. However, this risk adapted approach is still under prospective evaluation. PMID- 15820922 TI - [Practical use of o,p'DDD in adrenocortical carcinoma]. AB - Adrenocortical carcinoma (AC) is a rare tumor of poor prognosis. Its treatment by o,p'DDD remains a reference after initial surgery. Two galenic forms were recently available in France: Lysodren and Mitotane AP-HP. As Lysodren got a European registration in april 2004, Mitotane is no more produced. O,p'DDD is an adrenolytic and cytotoxic agent. It also reduces the hormonal secretion in AC. Its blood level must reach a therapeutic window (14-20 mg/l) to be effective and to limit toxicity. It is given orally three times a day (3 grams a day for Lysodren and 6-12 grams a day for Mitotane. Its posology is adapted according to serum levels of o,p'DDD and tolerance. Side effects are essentially gastrointestinal (GI), neurologic and hepatic. Five patients (four with AC and one with a metastatic Leydig cell tumor of the testis) were treated by Lysodren. Three patients had early Lysodren discontinuation due to toxicity (skin rash, weight loss, GI toxicity). Studies of combination with other treatments as chemotherapy and targeted drugs are warranted. Surgery is an important part of metastatic disease treatment. PMID- 15820923 TI - [Male breast cancer: about 123 cases collected at the Institute Salah-Azaiz of Tunis from 1979 to 1999]. AB - Male breast cancer is rare compared to its female counterpart representing less than 1% of cancer in men. The objective of our retrospective study is to report the epidemiologic and clinical profile and to analyse the therapeutic results and prognostic factors in a Tunisian population collected during a period of 20 years at a single institution. We collected from 1979 to 1999, all the histological confirmed male breast cancers treated at our institution. We analyse the following data: age, clinical presentation and features, therapeutic protocol, results and prognostic factors. Survival was done with the Kaplan-Meier method and comparison with the log-rank test. 123 cases of male breast carcinoma were collected with a median age of 65 years. Most patients (62.2%) have an advanced T4 disease with bilateral lesions in 4 cases. Infiltrating ductal carcinoma represent 91% of all tumours. 85% of tumours expressed hormonal receptor. The treatment consisted in a radical mastectomy in 93 cases (84%) followed by radiotherapy, chemotherapy and in many cases by hormonotherapy. After a median follow up of 26 month, 22 patients presented loco regional recurrence and 41 metastases. Estimated 5-year survival rate was 62%. The presence of metastasis, nodal involvement, advanced disease, and grade affected survival. Male breast cancer represent at our institution 1 % of the male cancers treated comparable to the literature data. T4 tumours represent a higher rate, the treatment approach is the standard applicable in breast cancer; prognostic factors are the classical one like breast cancer in women. PMID- 15820924 TI - [Evaluation of cancer treatments: What is its psychosocial impact?]. PMID- 15820925 TI - Altered expression of adhesion receptors after ex vivo short-term expansion of primitive progenitor cell. PMID- 15820926 TI - Efficacy and safety of sildenafil in the treatment of severe pulmonary hypertension in patients with hemoglobinopathies. PMID- 15820927 TI - Gene expression analysis fails to identify patients with chronic myeloid leukemia who will achieve cytogenetic response to imatinib. PMID- 15820928 TI - IgVH mutations in patients with chronic lymphocytic leukemia. PMID- 15820929 TI - Analysis of VH genes in 49 marginal zone lymphoma cases reveals marked heterogeneity between splenic and nodal tumors and suggests the existence of clonal selection. PMID- 15820930 TI - Adhesion of myeloma cells to the bone marrow stroma matrix protein fibronectin is stimulated by hepatocyte growth factor. PMID- 15820931 TI - CD44v6, a target for novel antibody treatment approaches, is frequently expressed in multiple myeloma and associated with deletion of chromosome arm 13q. PMID- 15820932 TI - Factor VIII half-life and intensity of treatment in hemophilic patients. AB - It is well known that severe hemophilic patients have a considerable different factor VIII half-lives. Details of the effect of the inter-individual variation of Factor VIII half-live on treatment or clinical outcome has not been reported. Accordingly, during many years several strategies in prophylactic treatment for severe haemophilia have been used (intermediate-dose of plasma FVIII versus high dose regime). In the retrospective study by van Dijk et al. (see page 494), the authors show an association between factor VIII half-live and the clinical characteristics of patients with severe hemophilia. Patients with shorter factor VIII half-life need more intensive treatment to prevent joint bleeds and arthropaty than patients with longer factor VIII half-life. Further prospective studies should necessary to confirm whether assessment of factor VIII half-life before the start of prophylaxis could modify the intensity of treatment and prevention of arthropaty among patients with severe hemophilia. PMID- 15820933 TI - Elevated levels of D-dimer and fragment 1+2 upon central venous catheter insertion and factor V Leiden predict subclavian vein thrombosis. AB - Symptomatic subclavian vein thrombosis is a potential complication in approximately 5-10% patients with indwelling central venous catheters. Having reliable tests that would identify those patients with a high risk of catheter related thrombosis would be clinically useful by helping to tailor thromboprophylaxis. In this case control study by Jansen et al. (see page 499), patients with a Hickman catheter who were undergoing allogeneic bone marrow transplantation for hematological malignancies were investigated. Baseline levels of D-dimer and fragment 1+2 were found to be significantly higher in patients who developed catheter-related thrombosis and the presence of factor V Leiden genotype was found only in patients with thrombosis. Although these abnormalities were associated with a 5- to 7-fold increased risk of catheter-related thrombosis, the positive predictive values for elevated D-dimer and fragment 1+2 levels were only 80% while the negative predictive values were about 40%. These results support the concept that laboratory evidence of activation of coagulation correlates with clinical thrombotic events, but they are insufficient for recommending routine testing of these markers in patients with central venous catheters. Further studies are required to confirm these findings and to see if similar results are observed in patients with other malignancies. PMID- 15820934 TI - A new oxygen transport agent. AB - Modern highly purified and chemically modified hemoglobin-based oxygen carriers (HBOC) are free of significant side effects on kidneys and coagulation, and they do not possess ABO antigens, allowing transfusion without knowledge of the respective blood group. Even at room air oxygen concentrations HBOC can compensate for intravascular volume deficits in hemorrhagic shock, including restoration of colloid osmotic pressure and organ perfusion, and deliver oxygen to organs and tissues during nearly complete blood exchange. In animal experiments and clinical trials all HBOC showed a vasoconstrictive side-effect which is mainly caused by nitric oxide scavenging, and to a lesser extent by reactive vasoconstriction because of precapillary oxygen off-loading. The study by Bjorkholm in this issue of the journal (see page 505) investigates the application of a moderate dose of the newly designed HBOC, MP4, in volunteers. MP4 has a high molecular size and a very low p50 resulting in a high oxygen affinity thus avoiding significant (pre)capillary oxygen off-loading. No significant rises in blood pressure or major laboratory abnormalities were seen after MP4 infusion. This new HBOC may be applicable in patients as a red blood substitute where vasoconstriction must be avoided. In addition, poststenotic tissue oxygenation might be a further indication. However, the number of treated volunteers and the infused dose of MP4 were both are very small. Therefore, one cannot draw conclusions on the safety, tolerability and efficacy of MP4 in terms of red cell replacement when large amounts of oxygen carriers are needed. PMID- 15820935 TI - Use of mesenchymal stem cells for the prevention of immune complications of hematopoietic stem cell transplantation. PMID- 15820936 TI - Rising CMV PP65 antigenemia and DNA levels during preemptive antiviral therapy. PMID- 15820937 TI - Pulmonary hypertension in patients with hemoglobinopathies: could a mechanism for dysfunction provide an avenue for novel therapeutics? PMID- 15820938 TI - Modulation of homing properties of primitive progenitor cells generated by ex vivo expansion. AB - BACKGROUND AND OBJECTIVES: The maintenance of adequate interactions with the bone marrow (BM) microenvironment is critical to ensure efficient homing of ex vivo expanded hematopoietic cells. This study was intended to assess adhesion and migration properties of long-term culture-initiating cells (LTC-IC) harvested after self-renewal division in ex vivo culture and to determine their susceptibility to growth-inhibitory signals mediated by adhesion to BM stromal ligands. DESIGN AND METHODS: We used cell tracking to isolate primitive LTC-IC that had accomplished 1 or 2 divisions ex vivo. Adhesion, migration and growth inhibition of divided LTC-IC were determined in the presence of purified BM ligands, and compared to the properties of uncultured LTC-IC. RESULTS: As compared to undivided LTC-IC, adhesion and migration mediated by very late antigen (VLA)-4 integrin across both vascular cell adhesion molecule-1 (VCAM-1) and fibronectin (Fn) were downregulated in post-mitotic LTC-IC. Conversely, binding and motility via VLA-5 across Fn were stimulated. No changes occurred in LTC-IC interactions with intercellular adhesion molecule-1 (ICAM-1) or with E- or P-selectin. Proliferation of uncultured LTC-IC was inhibited by VLA-4-mediated binding to VCAM-1 and the CS-1 domain of Fn, as well as binding to P-selectin. Growth of ex vivo-generated LTC-IC became unresponsive to these 3 ligands but was suppressed through VLA-5 engagement by the cell binding domain of Fn. INTERPRETATION AND CONCLUSIONS: The generation of LTC-IC in expansion culture is associated with functional alterations of adhesion receptors, modulating not only binding and migration in the BM but also responsiveness to adhesion-mediated growth inhibitory signals. Such changes may limit homing and engraftment of expanded primitive stem/progenitor cells. PMID- 15820939 TI - Efficacy and safety of sildenafil in the treatment of severe pulmonary hypertension in patients with hemoglobinopathies. AB - BACKGROUND AND OBJECTIVES: During the last decade new approaches to the treatment of pulmonary arterial hypertension (PH) have increased symptomatic relief and prolonged survival. PH is a common sequel of the hemoglobinopathies, thalassemia and sickle cell anemia, but the use of standard oral treatment options, such as calcium channel blockers, endothelin receptor antagonists, and long-term anticoagulation therapy, is limited because of toxicity and poor effectiveness. Sildenafil citrate is a selective and potent inhibitor of cGMP-specific phosphodiesterase-5 (PDE5) which promotes selective smooth muscle relaxation in lung vasculature and has been utilized successfully in the treatment of PH. The primary objective of this study was to evaluate the efficacy of sildenafil treatment in the control of PH in patients with hemoglobinopathies. DESIGN AND METHODS: In this study patients with hemoglobinopathies (thalassemia intermedia n=4; thalassemia major n=2; sickle thalassemia n=1) suffering from severe PH were treated with sildenafil citrate (50 mg b.i.d.) for periods ranging from 4 to 48 months. RESULTS: A significant decrease in pulmonary pressure and improvement in exercise capacity and functional class were observed in all patients. No significant adverse events were reported. INTERPRETATION AND CONCLUSIONS: These data, in a small group of patients, indicate that sildenafil citrate is effective in the treatment of PH in hemoglobinopathies that cannot be treated with alternative oral drugs and is well tolerated long-term at a daily dose of 100 mg, though studies including more patients may uncover toxicities and limitations of efficacy. PMID- 15820940 TI - In chronic myeloid leukemia white cells from cytogenetic responders and non responders to imatinib have very similar gene expression signatures. AB - BACKGROUND AND OBJECTIVES: Imatinib induces complete cytogenetic responses (CCR) in the majority of patients with chronic myeloid leukemia (CML) in chronic phase (CP). However, a subgroup of patients is refractory at the cytogenetic level. Clinically, it would be advantageous to identify such patients a priori, since they may benefit from more aggressive therapy. DESIGN AND METHODS: To elucidate mechanisms underlying cytogenetic refractoriness, we used Affymetrix oligonucleotide arrays to determine the transcriptional signature associated with cytogenetic refractoriness in unselected white blood or bone marrow cells from 29 patients with CML in first CP prior to treatment with imatinib. Patients with CCR within 9 months were defined as responders (n = 16) and patients lacking a major cytogenetic response (> 35% Philadelphia-positive metaphases) after 1 year were defined as non-responders (n = 13). RESULTS: Differences in gene expression between responders and non-responders were subtle. Stringent statistical analysis with multiple comparison adjustments revealed very few differentially expressed genes. Differentially expressed genes could not be confirmed in an independent test set. INTERPRETATION AND CONCLUSIONS: We conclude that transcriptional profiling of unselected white cells is of limited value for identifying genes consistently associated with cytogenetic refractoriness to imatinib. PMID- 15820941 TI - Patients with chronic lymphocytic leukemia with mutated VH genes presenting with Binet stage B or C form a subgroup with a poor outcome. AB - BACKGROUND AND OBJECTIVES: The immunoglobulin VH gene mutation status is a strong prognostic indicator in B-cell chronic lymphocytic leukemia (CLL), since unmutated VH genes are correlated with short survival. However, the traditional cut-off level dividing mutated and unmutated cases, i.e. more or less than 2% mutations, has been questioned and other cut-offs have been suggested. We investigated whether an alternative cut-off should be applied and the relation of mutational status to another prognostic marker, Binet staging. DESIGN AND METHODS: VH gene mutation status was assessed in 332 CLL cases by polymerase chain reaction amplification and nucleotide sequencing and was further correlated with overall survival using different VH mutation cut-offs (1-7%) and Binet stage. RESULTS: After testing different mutation borders, the 2% cut-off remained the best discriminative level for determining prognosis. Interestingly, prognostic stratification was improved by combining the information on VH gene mutation status with that of Binet stage: unmutated cases (all stages, n=151, mutated cases with stage A (n=77), and mutated cases with stage B or C (n=37) had a median survival of 82, 179 and 74 months, respectively. INTERPRETATION AND CONCLUSIONS: CLL cases displaying mutated VH genes with Binet stage B or C had a survival similar to that of unmutated cases and significantly shorter than that of mutated stage A CLL. Our result reveals clinical heterogeneity within the VH mutated CLL group by inclusion of Binet stage data, a finding which is of importance when considering surrogate marker(s) for VH mutation status. PMID- 15820942 TI - Analysis of VH genes in marginal zone lymphoma reveals marked heterogeneity between splenic and nodal tumors and suggests the existence of clonal selection. AB - BACKGROUND AND OBJECTIVES: To clarify the relationship between splenic (SMZL) and nodal marginal zone (NMZL) lymphomas, we analyzed immunoglobulin variable heavy chain (VH) gene usage and mutation patterns in these tumors. DESIGN AND METHODS: VH genes were cloned and sequenced from 49 lymphoma samples (35 SMZL and 14 NMZL). RESULTS: A biased usage of VH gene was found with overrepresentation of VH1 in SMZL cases (13/35) and VH4 in NMZL cases (7/14). Evidence for antigen driven mutations was identified in 8 SMZL and 4 NMZL cases. Three cases out of 18 with clones analyzed from spleen and peripheral blood demonstrated intra-clonal diversity, with evidence of clonal selection in one case, indicating the possibility of antigen-driven clonal expansion. Eleven SMZL cases (31%) but only 2 NMZL (14%) cases were unmutated. No differences in clinical outcome and overall survival were found between the unmutated and mutated cases. INTERPRETATION AND CONCLUSIONS: The pattern of somatic mutation and the VH gene segment usage appear to differ between SMZL and NMZL, suggesting that these are distinct pathological entities. Moreover, a biased usage of certain sequences suggests that tumor cells in SMZL may be subjected to antigen selection. PMID- 15820943 TI - Human myeloma cells adhere to fibronectin in response to hepatocyte growth factor. AB - BACKGROUND AND OBJECTIVES: Multiple myeloma is characterized by an accumulation of malignant plasma cells in the bone marrow. Inside the bone marrow, adhesion of myeloma cells to extracellular matrix proteins such as fibronectin may promote cell survival and induce drug resistance. In this work we examined the effect of hepatocyte growth factor (HGF) on the adhesion of myeloma cells and the signaling pathways involved. DESIGN AND METHODS: Cell adhesion experiments were performed with the human myeloma cell line INA-6 and primary myeloma cells. The HGF signaling pathway was studied in INA-6 cells with the use of antibodies against VLA-4 integrin, and with inhibitors of various intracellular signaling molecules. RESULTS: We found that HGF stimulated adhesion of myeloma cells to fibronectin. This event was dependent on the alpha4 and beta1 integrin subunits (VLA-4), but HGF did not increase the expression of integrins on the cell surface. Our findings suggest that HGF promotes myeloma cells to adhere via activation of the phosphatidylinositol 3-kinase (PI3K) pathway independently of AKT, but possibly through the involvement of nuclear factor kappa B (NF-kappaB). INA-6 cells adhered to fibronectin after stimulation by insulin-like growth factor or stromal cell-derived factor 1alpha, but this adhesion was less dependent on PI3K than HGF mediated adhesion. INTERPRETATION AND CONCLUSIONS: his work points to HGF as a pro-adhesive factor in cell adherence to the bone marrow matrix protein fibronectin, an event known to promote cancer cell survival and drug resistance. Inhibiting HGF, its receptor c-Met or the VLA-4 integrin may be beneficial to the myeloma patient. PMID- 15820944 TI - CD44v6, a target for novel antibody treatment approaches, is frequently expressed in multiple myeloma and associated with deletion of chromosome arm 13q. AB - BACKGROUND AND OBJECTIVES: Despite recent advances in the treatment of multiple myeloma (MM), this disease remains incurable in the majority of patients. Therefore, innovative treatment strategies are mandatory. Bivatuzumab mertansine is a novel cytotoxic immunoconjugate specifically targeting the CD44 splice variant CD44v6. To investigate the applicability of this compound to clonal plasma cell disorders, we analyzed CD44v6 expression on malignant plasma cells from patients with multiple myeloma. DESIGN AND METHODS: Bone marrow samples from 57 patients with monoclonal gammopathy of undetermined significance (MGUS), MM, and plasma cell leukemia (PCL) were examined for CD44v6 expression by using flow cytometry (FACS) analysis. In addition, all samples were investigated by fluorescence in situ hybridization (FISH) with a specific probe for the chromosomal band 13q14. RESULTS: In only 1 of 16 cases (6%) with MGUS and 1 out of 6 cases (17%) with stage I MM were plasma cells CD44v6 positive. In contrast, 43% of the cases with stage II/III MM or PCL expressed CD44v6. In these cases, CD44v6 expression was significantly correlated with chromosome 13q14 deletion as determined by FISH (p=0.02). INTERPRETATION AND CONCLUSIONS: CD44v6 is frequently expressed in advanced, high-risk MM. CD44v6 expression correlates with chromosomal band 13q14 deletions, a well-known risk factor in MM. These results suggest that this epitope is a potential new target for monoclonal antibodies such as bivatuzumab mertansine. PMID- 15820945 TI - Factor VIII half-life and clinical phenotype of severe hemophilia A. AB - BACKGROUND AND OBJECTIVES: Patients with severe hemophilia A have considerably different factor VIII half-lives. Whether this is associated with their clinical characteristics has not been reported. The aim of this study was to describe the association of factor VIII half-lives with treatment and clinical characteristics of patients with severe hemophilia A, who have been treated with individually tailored prophylaxis. DESIGN AND METHODS: Patients were selected from a single center cohort of 214 patients with severe hemophilia, born between 1944 and 1995. To improve efficiency we measured factor VIII half-life in 42 patients selected from the extremes of the distribution of phenotypes of severe hemophilia. We assessed information on life-long joint bleeds and clotting factor consumption. Orthopedic outcome was assessed by the Pettersson score. RESULTS: Among these patients with severe hemophilia, factor VIII half-life ranged from 7.4-20.4 hours (median 11.8 hours). A one-hour increase in factor VIII half-life was associated with 96 (95% confidence interval (CI) 2 -190) IU less clotting factor use per kg per year. Age was an important determinant of factor VIII half-life, and explained a large part of the association between factor VIII half-life and clotting factor consumption. The median number of joint bleeds per year and arthropathy were similar for patients with different half-lives. INTERPRETATION AND CONCLUSIONS: Among patients with severe hemophilia, treated prophylactically with clotting factor, those with a shorter factor VIII half-life required slightly more clotting factor to prevent joint bleeds and subsequent arthropathy than similar patients with a longer factor VIII half-life. PMID- 15820946 TI - Elevated levels of D-dimer and fragment 1+2 upon central venous catheter insertion and factor V Leiden predict subclavian vein thrombosis. AB - BACKGROUND AND OBJECTIVES: Subclavian vein thrombosis is a well-recognized complication following central venous catheter insertion. We studied whether the determination of D-dimer levels, fragment 1+2 levels and factor V Leiden can identify patients at high risk of developing subclavian vein thrombosis. DESIGN AND METHODS: The presence of central venous catheter associated thrombosis was analyzed in 235 patients undergoing allogeneic bone marrow transplantation, of whom 30 (13%) developed thrombosis. A case-control study was performed with 30 patients matched for age, gender, and type of transplantation who did not develop thrombosis. Blood was sampled 3-5 days after catheter insertion. D-dimer levels were determined using a latex microparticle assay and an enzyme linked immunosorbent assay (ELISA). An ELISA was used to determine fragment 1+2 levels. The factor V genotype was determined by polymerase chain reaction. RESULTS: The levels of D-dimer and fragment 1+2 were significantly elevated in the patients who developed thrombosis. Five patients tested positive for factor V Leiden and all 5 developed subclavian vein thrombosis. Patients with high D-dimer levels (> 1300 microg/L measured by latex agglutination and >350 microg/L measured by ELISA) had a 7.0 and 6.0 times higher risk of developing subclavian vein thrombosis, respectively. A 5.5-fold increased risk of thrombosis was observed in patients with a fragment 1+2 level higher than 1.300 nmol/L. This resulted in positive predictive values of 0.78, 0.80 and 0.83 for the fragment 1+2, D-dimer and D-dimer latex agglutination assays, respectively. The accompanying negative predictive values were 0.39, 0.40 and 0.42, respectively. INTERPRETATION AND CONCLUSIONS: We conclude that the measurement of D-dimer and fragment 1+2 levels after central venous catheter insertion, as well as factor V Leiden determination, can be used to identify patients at high risk of developing symptomatic subclavian vein thrombosis. PMID- 15820947 TI - A phase I single blind clinical trial of a new oxygen transport agent (MP4), human hemoglobin modified with maleimide-activated polyethylene glycol. AB - BACKGROUND AND OBJECTIVES: MP4 (Hemospan), a hemoglobin-based oxygen carrier, has been designed to deliver oxygen to hypoxic tissues without causing vasoconstriction. A phase I clinical trial of MP4 was undertaken to evaluate whether MP4 elicits the clinical side effects associated with previous hemoglobin based solutions. DESIGN AND METHODS: Twelve volunteers were studied. One cohort (n=4) received 50 mg/kg of MP4, a second (n=4) received 100 mg/kg of MP4, and the third (n=4) received lactated Ringer's solution. Single blind infusions were given at 5 mL/min. Vital signs and symptoms, hematologic parameters, serum chemistry, renal and electrocardiographic measurements were monitored for 15 days after dosing. RESULTS: Five mild adverse events occurred in the controls and 2 each in the 50 mg/kg and 100 mg/kg MP4 groups. None was severe or judged related to MP4 administration by the principal investigator. There were no clinically significant alterations in blood pressure or heart rate, and there were no gastrointestinal symptoms, abdominal or flank pain, loss of appetite or clinically significant alterations of liver or pancreatic enzymes. In one subject (100 mg/kg of MP4), amylase and lipase were slightly above the upper limit of normal 4 hours after dosing, but without associated symptoms or signs. Pharmacokinetic analysis of plasma hemoglobin (assuming no hemolysis) yielded an estimated half-life (T1/2) of 43 hours in the 100 mg/kg MP4 subjects. INTERPRETATION AND CONCLUSIONS: MP4 appears to have a favorable safety profile. Subjects in both study groups survived and did no less well than those in the control group. PMID- 15820948 TI - Interaction of human mesenchymal stem cells with cells involved in alloantigen specific immune response favors the differentiation of CD4+ T-cell subsets expressing a regulatory/suppressive phenotype. AB - BACKGROUND AND OBJECTIVES: Experimental evidence and preliminary clinical studies have demonstrated that human mesenchymal stem cells (MSC) have an important immune modulatory function in the setting of allogeneic hematopoietic stem cell (HSC) transplantation. We extended the evaluation of mechanisms responsible for the immune regulatory effect derived from the interaction of human MSC with cells involved in alloantigen-specific immune response in mixed lymphocyte culture (MLC). DESIGN AND METHODS: Dendritic cell (DC) differentiation, T- and natural killer (NK)-lymphocyte expansion, alloantigen-specific cytotoxic activity and differentiation of CD4+ T-cell subsets co-expressing CD25 and/or CTLA4 molecules were assessed, comparing the effect observed using third-party MSC with that obtained employing MSC autologous to the MLC responder. RESULTS: We found that human MSC strongly inhibit alloantigen-induced DC1 differentiation, down-regulate alloantigen-induced lymphocyte expansion, especially that of CD8+ T cells and of NK lymphocytes, decrease alloantigen-specific cytotoxic capacity mediated by either cytotoxic T lymphocytes or NK cells and favor the differentiation of CD4+ T-cell subsets co-expressing CD25 and/or CTLA4. More effective suppressive activity on MLC-induced T-cell activation was observed when MSC were third-party, rather than autologous, with respect to MLC-responder cells. INTERPRETATION AND CONCLUSIONS: Our results strongly suggest that MSC-mediated inhibition of alloantigen-induced DC1 differentiation and preferential activation of CD4+ CD25+ T-cell subsets with presumed regulatory activity represent important mechanisms contributing to the immunosuppressive activity of MSC. Collectively, these data provide immunological support for the use of MSC to prevent immune complications related to both HSC and solid organ transplantation and to the theory that MSC are universal suppressors of immune reactivity. PMID- 15820949 TI - Rising antigenemia levels may be misleading in pre-emptive therapy of human cytomegalovirus infection in allogeneic hematopoietic stem cell transplant recipients. AB - BACKGROUND AND OBJECTIVES: Hematopoietic stem cell transplant (HSCT) recipients after show rising levels of antigenemia during pre-emptive ganciclovir treatment of human cytomegalovirus (HCMV) infection. This raises some doubts about the therapeutic decisions to be taken. DESIGN AND METHODS: Three groups of HSCT recipients with HCMV infection undergoing anti-viral treatment were identified: group A, showing increasing antigenemia and decreasing viremia and DNAemia; group B, with simultaneous increases in antigenemia, viremia, and DNAemia; and group C, with decreasing levels of all 3 viral markers. Viral load, determined as levels of antigenemia, viremia and DNAemia, was monitored for 3 months post transplantation in all groups. RESULTS: Group A HSCT recipients showed antigenemia peaks 2-11 days after the onset of treatment, reaching negative levels only 25-30 days thereafter, whereas viremia and DNAemia started to drop earlier. Group B patients, mainly including HSCT recipients with grade II-IV acute GvHD treated with steroids prior to and during antiviral treatment, showed increasing levels of all three viral parameters until 5-10 days after the start of treatment; the levels dropped to negative values 25-30 days thereafter. Group C patients, who acted as controls, progressively cleared virus from blood as an early result of antiviral therapy. INTERPRETATION AND CONCLUSIONS: Antigenemia is not the best assay to guide pre-emptive therapy. Group A patients, who have an isolated increase of antigenemia, do not require a change of the ongoing antiviral therapy. Whether better control of infection could be obtained in group B patients by either reducing immunosuppressive therapy (when possible) or adopting combination therapy remains to be determined. PMID- 15820950 TI - New tyrosine kinase inhibitors in chronic myeloid leukemia. AB - The deregulated activity of BCR-ABL tyrosine kinase originating from the t(9;22) chromosomal translocation has been shown to be necessary and sufficient for the transformed phenotype of chronic myeloid leukemia (CML) cells. This peculiarity has paved the way for the development of novel therapies specifically targeting the BCR-ABL gene product. The first BCR-ABL inhibitor to come into use in clinical practice, imatinib mesylate, is now the first-choice treatment for all newly diagnosed CML patients, but the initial striking efficacy of this drug has been overshadowed by the development of clinical resistance. The most common mechanisms of resistance include (i) BCR-ABL overexpression, and (ii) BCR-ABL kinase domain mutations disrupting critical contact points between imatinib and BCR-ABL or inducing a transition to a conformation to which imatinib is unable to bind. Several approaches to overcoming resistance have been studied both in vitro and in vivo. They include dose escalation of imatinib, the combination of imatinib with chemotherapeutic drugs, alternative BCR-ABL inhibitors, and inhibitors of kinases acting downstream of BCR-ABL such as Src kinases. Various novel tyrosine kinase inhibitors (TKI) have been synthesized and have now reached the pre-clinical or clinical phase. This review highlights the development of new TKI as specific molecularly targeted therapy and as the principal mechanisms for overcoming imatinib resistance. PMID- 15820951 TI - Prions and the blood and immune systems. AB - Prion diseases take a number of forms in animals and humans. They are caused by conformational change in widely expressed prion protein leading to the formation of intracellular aggregates. Although the main focus of disease is the central nervous system, it is known that involvement of the immune system occurs in peripherally transmitted disease in particular. Animal experiments suggest that in some prion diseases follicular dendritic cells in the germinal centers are a major site of initial accumulation, and that abnormal prion protein and infectivity are detectable in peripheral lymphoid tissue from the earliest phase of disease. This raises the possibility that in a human peripherally transmitted prion disease like variant Creutzfeldt-Jakob disease, further transmission could occur through blood or tissue products or contamination of surgical instrumentation. Indeed two recent reports confirm that this disease has been transmitted by blood, raising significant public health concerns. PMID- 15820952 TI - Mature erythrocyte indices: new markers of iron availability. AB - This study was aimed at evaluating mature erythrocyte indices as new markers of iron status. Contrarily to those in the whole red blood cell (RBC) population, mature erythrocyte parameters are valid markers of iron status that remain independent of erythropoietic activity. When reticulocytosis is low, these parameters are similar to whole RBC parameters. PMID- 15820953 TI - A new polyadenylation site mutation associated with a mild beta-thalassemia phenotype. AB - At least 180,000 autochthonous and allochthonous people are carriers of a large spectrum of hemoglobinopathies in The Netherlands. We describe two cases, the first, a 33-year old Surinamese Creole woman, studied because of an intermediate hemolytic anemia; the second, a couple requesting analysis because of a previously diagnosed carrier state in the male partner, while the carrier state in the pregnant female was uncertain. PMID- 15820954 TI - Nutritional iron status in children with alpha+ thalassemia and the sickle cell trait in a malaria endemic area on the coast of Kenya. AB - Although hemoglobinopathies such as alpha+ thalassemia and the sickle cell trait might contribute to anemia in African children, we hypothesized that they might also enhance iron absorption under circumstances of critical availability, and that this could attenuate their hematologic effects. We found no support for this hypothesis in a cohort of children living on the coast of Kenya. PMID- 15820955 TI - Neutropenia in Iranian patients with primary immunodeficiency disorders. AB - Neutropenia may occur in some of the primary immunodeficiency disorders. We reviewed the records of 56 neutropenic patients. The most common disorders were Shwachman-Diamond syndrome, cyclic neutropenia, Kostmann disease, Chediak-Higashi syndrome, hyper IgM syndromes, severe combined immunodeficiency, hyper IgE syndrome, and common variable immunodeficiency. PMID- 15820957 TI - The incidence of submicroscopic deletions in reciprocal translocations is similar in acute myeloid leukemia, BCR-ABL positive acute lymphoblastic leukemia, and chronic myeloid leukemia. AB - We compared the incidence of submicroscopic deletions accompanying balanced translocations using interphase fluorescence in situ hybridization (FISH) in 245 patients with chronic myeloid leukemia (CML), 79 patients with acute lymphoblastic leukemia (ALL) and BCR-ABL (n=70) or MLL rearrangements (n=29), and 412 patients with acute myeloid leukemia (AML) with CBFB-MYH11 (n=122), PML RARalpha (n=108), AML1-ETO (n=112), or MLL rearrangements (n=98). The incidence of submicroscopic deletions was 2-9% depending on the entity. PMID- 15820956 TI - Clonal cytogenetic abnormalities in patients with chronic myeloid leukemia in complete cytogenetic response to imatinib mesylate. AB - The emergence of clonal chromosomal abnormalities in Philadelphia-negative cells during treatment with imatinib in patients with Philadelphia-positive chronic myeloid leukemia has been reported. We add information to this issue presenting a series of 29 patients in complete cytogenetic response after imatinib treatment, three of whom developed clonal aberrations. PMID- 15820958 TI - Global real-time quantification/reverse transcription-polymerase chain reaction for detecting proto-oncogenes associated with 14q32 chromosomal translocation in multiple myeloma. AB - A global real-time quantitative/reverse transcription-polymerase chain reaction technique for detecting the expression of six 14q32 chromosomal translocation associated proto-oncogenes in marrow plasma cells was established and applied to myeloma specimens. This technique is an alternative method of detecting 14q32 rearrangements and allows investigation of the relationship between proto oncogene expression and clinical features. PMID- 15820959 TI - An improved method for lupus anticoagulant detection. AB - The diagnosis of lupus anticoagulant (LA) becomes difficult when there is a weak titer of LA antibodies. This study demonstrates that 20 minutes' incubation increases the sensitivity of the mixing test to diagnose LA by 23% and to suspect the antibodies by 66%. PMID- 15820960 TI - Double versus single autotransplantation in multiple myeloma; a single center experience of 100 patients. AB - One hundred patients with newly diagnosed multiple myeloma (MM) were treated with high-dose chemotherapy followed by single or double autologous stem cell transplantation (ASCT). Up-front treatment with a double ASCT tended to prolong progression-free and overall survival. PMID- 15820961 TI - Long-term follow-up of lymphocyte populations and cellular cytokine production in patients with chronic graft-versus-host disease treated with extracorporeal photopheresis. AB - We studied lymphocyte populations and cytokine-expression profiles of ten patients with chronic graft-versus-host disease who at least transiently responded to photoimmunotherapy. The numbers of lymphocytes, monocytes and dendritic cells rose in most cases. Th1 cells always increased during therapy, supporting the hypothesis that a more favorable immune balance contributes to clinical responses. PMID- 15820962 TI - Clinical grading of oral chronic graft-versus-host disease in 104 consecutive adult patients. AB - The aim of this study was to investigate the clinical relevance of oral involvement by chronic graft-versus-host disease. The presence of oral changes in association with skin and other target organs including eye, lung or joint may adversely influence the probability of discontinuing systemic immunosuppressive treatment. PMID- 15820963 TI - Cytomegalovirus infection and chronic graft-versus-host disease are significant predictors of renal failure after allogeneic hematopoietic stem cell transplantation. AB - We investigated the frequencies and predictive factors of prolonged renal failure (PRF) in a retrospective study of 181 consecutive patients undergoing allogeneic hematopoietic stem cell transplantation. Twenty-six patients (23% of long-term survivors) developed PRF. We identified 4 independent prognostic factors; cytomegalovirus infection and chronic graft-versus-host disease appeared as major risk factors for PRF. PMID- 15820964 TI - A psychosocial assessment interview of candidates for hematopoietic stem cell transplantation. AB - There is evidence that psychosocial issues are associated with poorer compliance and higher mortality in the post-transplant period. Thus, psychosocial assessment of candidates for transplantation is an important way of detecting early those patients who are susceptible to developing psychiatric symptoms and psychosocial difficulties during treatment. We developed a psychosocial structured interview to assess candidates for hematopoietic stem cell transplantation. The interview is a short and comprehensible instrument, requiring an average of 50 minutes to be completed. PMID- 15820965 TI - No relationship between hepatitis C infection and risk of myeloid malignancy. AB - The etiology of myeloid malignancies is related to chromosomal damage caused by irradiation, alkylating agents, viral infection and so forth; however, few clinical studies have been published on the association between viral infection and the development of myeloid malignancies. PMID- 15820967 TI - The natural history of asymptomatic primary biliary cirrhosis. PMID- 15820966 TI - Modified Mediterranean diet and survival: EPIC-elderly prospective cohort study. AB - OBJECTIVE: To examine whether adherence to the modified Mediterranean diet, in which unsaturates were substituted for monounsaturates, is associated with longer life expectancy among elderly Europeans. DESIGN: Multicentre, prospective cohort study. SETTING: Nine European countries (Denmark, France, Germany, Greece, Italy, the Netherlands, Spain, Sweden, United Kingdom). PARTICIPANTS: 74,607 men and women, aged 60 or more, without coronary heart disease, stroke, or cancer at enrolment and with complete information about dietary intake and potentially confounding variables. MAIN OUTCOME MEASURES: Extent of adherence to a modified Mediterranean diet using a scoring system on a 10 point scale, and death from any cause by time of occurrence, modelled through Cox regression. RESULTS: An increase in the modified Mediterranean diet score was associated with lower overall mortality, a two unit increment corresponding to a statistically significant reduction of 8% (95% confidence interval 3% to 12%). No statistically significant evidence of heterogeneity was found among countries in the association of the score with overall mortality even though the association was stronger in Greece and Spain. When dietary exposures were calibrated across countries, the reduction in mortality was 7% (1% to 12%). CONCLUSION: The Mediterranean diet, modified so as to apply across Europe, was associated with increased survival among older people. PMID- 15820968 TI - Benign paroxysmal positional vertigo: clinical characteristics of dizzy patients referred to a Falls and Syncope Unit. AB - BACKGROUND: Dizziness is a common symptom in older people that affects quality of life and increases the risk of falls. Benign paroxysmal positional vertigo (BPPV) is a common cause of dizziness that increases in prevalence with age, and is potentially curable. AIM: To compare patients with BPPV referred initially to a Falls and Syncope Unit (FSS group) with those initially referred to a Regional ENT/Balance Service (ENT group). DESIGN: Retrospective case-note review. METHODS: Medical notes, investigations and outcomes were reviewed for all patients. RESULTS: Of 59 patients with BPPV confirmed by Dix-Hallpike test, 31 (53%) were initially referred to the FSS (2.6 patients per month, 71% females) and 28 (47%) were initially referred to ENT (4.7 patients per month, 86% females). Compared to those referred initially to ENT, FSS patients were significantly older (mean +/- SD 69 +/- 13 vs. 55.4 +/- 13 years, p = 0.0003) and had dizzy symptoms for longer before diagnosis (median (range) 12 (4-120) vs. 6 (1-36) months, p = 0.0273). FSS patients were more likely to have more than one type of dizziness (16% vs. 0%, p = 0.001), more likely to have cerebrovascular or cardiovascular co-morbidity (13% vs. 4%, p = 0.0152) and were taking significantly more medications (3.2 vs. 1.7; p = 0.0271). Cure rates on intervention were similar (83% FSS, 86% ENT). DISCUSSION: BPPV is a potentially curable cause for dizziness in older people. Older people are frequently referred directly to Falls units, who will be seeing increasing numbers of patients with dizziness. A high index of suspicion allows early identification and treatment of this condition. PMID- 15820969 TI - Pitfalls and problems of relying on serum troponin. AB - Cardiac troponin (cT) is released after myocardial damage. In the appropriate clinical setting, a measured elevation of cT can increase the diagnostic rate of myocardial infarction and acute coronary syndrome. Elevations of cT, however, can occur in a wide variety of other clinical situations. Failure to recognize this can lead to an over-diagnosis of myocardial infarction (MI). We present clinical cases from our institution that illustrate this diagnostic problem, and review similar cases in the literature. We also discuss the implications of an erroneous diagnosis of myocardial infarction, for the patient and for the health services. PMID- 15820970 TI - Castration reduces platelet thromboxane A2 receptor density and aggregability. AB - BACKGROUND: Exogenously administered testosterone upregulates platelet thromboxane A2 (TXA2) receptors and increases aggregation response to thromboxane mimetics in healthy male volunteers. However, the biological impact of endogenous testosterone on platelet TXA2 receptor expression, especially in older men at risk of coronary artery disease, is unclear. AIM: To investigate the impact of reduction in circulating testosterone on platelet TXA2 receptor expression in older men. DESIGN: Cross-sectional case-control study. METHODS: We studied surgically and/or medically castrated men with prostate cancer (group A, n = 8, aged 71 +/- 8 years) and age-matched, uncastrated urology patients (group B, n = 7, aged 67 +/- 9 years). Plasma testosterone was measured by radioimmunoassay. Platelet TXA2 receptor expression was assessed by radioligand binding studies using radioactive 125I-BOP. Platelet aggregation responses to TXA2-mimetic I-BOP, and to thrombin, were also studied. RESULTS: Group A had significantly lower plasma testosterone than group B (16 +/- 5 ng/dl vs. 308 +/- 47 ng/dl, p<0.001). Platelet TXA2 receptor density (B(max)) but not affinity (K(d)) was lower in group A (0.50 +/- 0.12 vs. 1.01 +/- 0.17 pmol/mg protein, p = 0.03). Maximum platelet aggregation response to I-BOP (E(max)), but not sensitivity (EC50) was lower in group A (53 +/- 2% vs. 63 +/- 2%, p = 0.003 ANOVA). In vitro, high concentrations of hydroxyflutamide (100 microM) competitively inhibited U46619 induced platelet aggregation in washed platelets, without affecting the binding of 125I-BOP to platelet TXA2 receptors. DISCUSSION: Endogenous testosterone regulates platelet TXA2 receptor B(max) and the E(max) aggregation response to thromboxane mimetic I-BOP. Blockade of androgen receptors or inhibition of testosterone production may reduce platelet aggregation responses. Preliminary evidence suggests the presence of functional androgen receptors on human platelets, which may regulate TXA2 receptor expression. PMID- 15820971 TI - Incidence and risk of arm oedema following treatment for breast cancer: a three year follow-up study. AB - BACKGROUND: Breast-cancer-related lymphoedema is a chronic condition with estimates of incidence ranging from 6 to 83%. Lymphoedema has been associated with a variety of risk factors. However, this evidence has suffered from methodological weaknesses, and so has had little impact upon clinical practice. AIM: To examine incidence and risk factors [hospital skin puncture, surgical procedure, Body Mass Index (BMI), age, axillary node status, number of axillary nodes removed, radiotherapy and surgery on dominant side] for breast cancer related arm lymphoedema. DESIGN: Prospective observational study, with measurement of limbs pre-operatively and at regular intervals post-operatively. METHODS: We recruited 251 women who had surgical treatment for breast cancer that involved sampling, excision or biopsy of axillary nodes, aged > or = 18 years, and free of advanced disease and psychological co-morbidities. Of these, 188 (74.9%) were available for 3-year follow-up. RESULTS: At follow-up, 39 (20.7%) had developed lymphoedema. Hospital skin puncture (vs. none) (RR 2.44, 95%CI 1.33 4.47), mastectomy (vs. wide local excision or lumpectomy) (RR 2.04, 95%CI 1.18 3.54), and BMI > or = 26 (vs. BMI 19-26) (RR 2.02, 95%CI 1.11-3.68) were the only significant risk factors. DISCUSSION: Lymphoedema remains a significant clinical problem, with 1:5 women in this sample developing the condition following treatment for breast cancer. Risk factors are identified in the development of lymphoedema that should be taken into account in clinical practice. PMID- 15820972 TI - Usage of troponin in the real world: a lesson for the introduction of biochemical assays. AB - BACKGROUND: Studies have demonstrated economic and clinical effectiveness using troponin as a risk stratification tool in chest pain patients. Those with a positive result are investigated invasively, whilst those with a negative result and ECG are promptly mobilized, facilitating discharge. AIM: To determine whether our use of troponin I (cTnI) in routine clinical practice conforms to ideal standards. DESIGN: Audit study. METHODS: Data were collected from 93 laboratory request forms for cTnI measurement on 72 patients with matched available patient records. RESULTS: Eighty requests had no information regarding timing of blood sample in relation to the clinical event; 39% gave no clinical indication. Only 71% of results were available within 12 h. An admission diagnosis of acute coronary syndrome (ACS) was made in 25%. Fifteen had typical cardiac chest pain with a negative cTnI: 6 of these had an exercise treadmill test before discharge. Nine had a positive cTnI, but only two had coronary angiography. Of patients with negative cTnI and possible ACS, 84% were in hospital for >4 days. DISCUSSION: The introduction of troponin assays into widespread use requires careful assessment. cTnI requests and subsequent patient management remain below expected standards. Ideally, the laboratory should provide an accurate result within a reasonable time frame, while physicians need to request cTnI at a suitable time-point and use the result appropriately. Lessons from the introduction of cTnI measurement may be useful for the introduction of future new tests in other areas of cardiology and medicine. PMID- 15820973 TI - Type D personality: the heart, stress, and cortisol. AB - Many studies have demonstrated the role of psychosocial and behavioural risk factors in the aetiology and pathogenesis of cardiovascular disorders. Recently, a new personality construct, the type D or 'distressed' personality, has been proposed. Type D behaviour is characterized by the joint tendency to experience negative emotions and to inhibit these emotions while avoiding social contacts with others. The observation that cardiac patients with type D personality are at increased risk for cardiovascular morbidity and mortality underlines the importance of examining both acute (e.g. major depression) and chronic (e.g. certain personality features) factors in patients at risk for coronary events. Both type D dimensions (negative affectivity and social inhibition) are associated with greater cortisol reactivity to stress. Elevated cortisol may be a mediating factor in the association between type D personality and the increased risk for coronary heart disease and, possibly, other medical disorders. Studies of the effect of age on hypothalamic-pituitary-adrenal (HPA) function in healthy humans have produced inconsistent results. This may relate to a different prevalence of type D individuals in study samples (i.e. some type D individuals may have alterations within the HPA axis that are similar to HPA axis changes in depressed patients). Further studies of the psychological and biological features of type D individuals may help develop treatment approaches to improve the psychological and physical health of individuals with type D personality. PMID- 15820974 TI - Treatment of multiple sclerosis: beyond the NICE guidelines. AB - Multiple sclerosis (MS) is a common, disabling neurological condition whose pathogenesis is not clearly understood. Although current treatment recommendations assume an immunopathogenic disease mechanism, MS may not be an autoimmune disorder. Long-term immunological therapy for MS is in our view an untested approach, guided by uncritical acceptance of data from drug trials. We do not believe that there is convincing evidence that any of these immune-based treatments prevents long-term disease progression, or has much effect on common disabilities such as fatigue, pain, depression and cognitive impairment. The recent recommendations of the National Institute of Clinical Excellence did not address important issues regarding disease modification, management of paroxysmal symptoms and the likely therapeutic candidates for future treatment trials. We discuss treatment options for MS beyond the NICE guidelines. PMID- 15820975 TI - Homology modelling of transferrin-binding protein A from Neisseria meningitidis. AB - Neisseria meningitidis, a causative agent of bacterial meningitis, obtains transferrin-bound iron by expressing two outer membrane located transferrin binding proteins, TbpA and TbpB. TbpA is thought to be an integral outer membrane pore that facilitates iron uptake. Evidence suggests that TbpA is a useful antigen for inclusion in a vaccine effective against meningococcal disease, hence the identification of regions involved in ligand binding is of paramount importance to design strategies to block uptake of iron. The protein shares sequence and functional similarities to the Escherichia coli siderophore receptors FepA and FhuA, whose structures have been determined. These receptors are composed of two domains, a 22-stranded beta-barrel and an N-terminal plug region that sits within the barrel and occludes the transmembrane pore. A three dimensional TbpA model was constructed using FepA and FhuA structural templates, hydrophobicity analysis and homology modelling. TbpA was found to possess a similar architecture to the siderophore receptors. In addition to providing insights into the highly immunogenic nature of TbpA and allowing the prediction of potentially important ligand-binding epitopes, the model also reveals a narrow channel through its entire length. The relevance of this channel and the spatial arrangement of external loops, to the mechanism of iron translocation employed by TbpA is discussed. PMID- 15820976 TI - Directed evolution of PDZ variants to generate high-affinity detection reagents. AB - High-throughput protease assays are used to identify new protease inhibitors which have the potential to become valuable therapeutic products. Antibodies are of great utility as affinity reagents to detect proteolysis products in protease assays, but isolating and producing such antibodies is unreliable, slow and costly. It has been shown previously that PDZ domains can also be used to detect proteolysis products in high-throughput homogeneous assays but their limited natural repertoire restricts their use to only a few peptides. Here we show that directed evolution is an efficient way to create new PDZ domains for detection of protease activity. We report the first use of phage display to alter the specificity of a PDZ domain, yielding three variants with up to 25-fold increased affinity for a peptide cleavage product of HIV protease. Three distinct roles are assigned to the amino acid substitutions found in the selected variants of the NHERF PDZ domain: specific 'beta1-beta3' interaction with ligand residue -1, interactions with ligand residues -4 to -7 and improvement in phage display efficiency. The variants, having affinities as high as 620 nM, display improvements in assay sensitivity of over 5-fold while requiring smaller amounts of reagents. The approach demonstrated here leads the way to highly sensitive reagents for drug discovery that can be isolated more reliably and produced less expensively. PMID- 15820977 TI - Conservation and specialization in PAS domain dynamics. AB - The PAS (Per-ARNT-Sim) superfamily is presented as a well-suited study case to demonstrate how comparison of functional motions among distant homologous proteins with conserved fold characteristics may give insight into their functional specialization. Based on the importance of structural flexibility of the receptive structures in anticipating the signal-induced conformational changes of these sensory systems, the dynamics of these structures were analysed. Molecular dynamics was proved to be an effective method to obtain a reliable picture of the dynamics of the crystal structures of HERG, phy3, PYP and FixL, provided that an extensive conformational space sampling is performed. Other reliable sources of dynamic information were the ensembles of NMR structures of hPASK, HIF-2alpha and PYP. Essential dynamics analysis was successfully employed to extract the relevant information from the sampled conformational spaces. Comparison of motion patterns in the essential subspaces, based on the structural alignment, allowed identification of the specialized region in each domain. This appears to be evolved in the superfamily by following a specific trend, that also suggests the presence of a limited number of general solutions adopted by the PAS domains to sense external signals. These findings may give insight into unknown mechanisms of PAS domains and guide further experimental studies. PMID- 15820978 TI - Improvement of Fc-erythropoietin structure and pharmacokinetics by modification at a disulfide bond. AB - Erythropoietin (Epo) is a cytokine that controls the production of red blood cells (RBCs). Epo acts continuously on RBC precursors to prevent apoptosis, so it is important to maintain high levels of Epo activity when treating anemic patients. We describe here modified human Epo [Epo(NDS)] with mutations His32Gly, Cys33Pro, Trp88Cys and Pro90Ala that result in the rearrangement of the disulfide bonding pattern from Cys29-Cys33 to Cys29-Cys88 and that, in the context of an Fc Epo(NDS) fusion protein, lead to significantly improved properties. Fc-Epo was secreted from NS/0 myeloma cells as about 35% high molecular weight aggregates, was unstable upon removal of N-linked oligosaccharides and showed poor pharmacokinetics and little efficacy in mice. In contrast, a corresponding Fc Epo(NDS) was secreted almost exclusively as a unit dimer, was relatively stable to removal of N-linked oligosaccharides, had much improved pharmacokinetic properties and had a significantly improved effect on RBC production. These results indicate that rearrangement of the disulfide bonding pattern in a therapeutic protein can have a significant effect on pharmacokinetics and, potentially, the dosing schedule of a protein drug. PMID- 15820979 TI - Detection of subunit interfacial modifications by tracing the evolution of clamp loader complex. AB - The archaeal and eukaryal clamp-loader and clamp proteins were investigated with the evolutionary trace method. The molecular phylogeny of the proteins suggested that the hetero-pentameric complex of the archaeal clamp-loader with two subunits (RFCL and RFCS) was not a preserved ancestral type, but a degenerated version of the eukaryal complex of five subunits (RFC1-5). The evolutionary trace of amino acid replacements during the course of subunit differentiation revealed that the replacements had accumulated preferentially at the subunit interface regions. Some of the interfacial modifications that might be responsible for the specific interaction between the subunits were conserved in the current complex. PMID- 15820980 TI - Computationally designed variants of Escherichia coli chorismate mutase show altered catalytic activity. AB - Computational protein design methods were used to predict five variants of monofunctional Escherichia coli chorismate mutase expected to maintain catalytic activity. The variants were tested experimentally and three active site mutants exhibited catalytic activity similar to or greater than the wild-type enzyme. One mutant, Ala32Ser, showed increased catalytic efficiency. PMID- 15820982 TI - Modeling membrane proteins based on low-resolution electron microscopy maps: a template for the TM domains of the oxalate transporter OxlT. AB - The availability of both EM and high-resolution crystallographic data for several membrane proteins (MPs) permits a detailed evaluation of the ability of molecular modeling techniques to complement EM data in the development of models of MPs. A protocol for this purpose is presented, consisting of (1) identifying transmembrane (TM) domains from sequence; (2) assigning buried and lipid-exposed faces of the TM domains; and (3) assembling the TM domains into a bundle, based on geometric restraints obtained from the EM data. The protocol is validated by predicting the structures of several 7- and 12-TM MPs to within 3-5 A r.m.s.d. from their crystal structures. The protocol is applied to generate a model of the oxalate transporter OxlT, for which a high-resolution structure is not yet available. PMID- 15820981 TI - A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells. AB - The HIV-1 TAT peptide has been used extensively for directing the intracellular delivery of an assortment of cargo, including DNA, liposomes and macromolecules. For protein delivery, a variety of TAT-fusion proteins have been described which link the TAT coding sequence to the protein coding sequence of interest. Streptavidin represents a potentially useful TAT-fusion protein because it could be used to deliver a wide array of biotinylated cargo. Here we have characterized a TAT-streptavidin (TAT-SA) fusion protein, which retains the ability to bind biotinylated cargo while directing their efficient cellular uptake. Fluorescence activated cell sorting (FACS) analysis and confocal microscopy characterization showed that TAT-SA is internalized by Jurkat T-cells and NIH 3T3 cells alone and when complexed to phycoerythrin, whereas the native streptavidin is not. Additionally, biotinylated alkaline phosphatase is successfully internalized and retains its activity when complexed to TAT-SA and incubated with Jurkat T-cells. Confocal microscopy suggested, however, that internalized TAT-SA and TAT-SA complexes were largely compartmentalized in vesicular compartments, rather than freely diffusing in the cytoplasmic compartment. To effect cytoplasmic delivery, the endosomal releasing polymer, poly(propylacrylic acid) (PPAA), was biotinylated and complexed to TAT-SA. Endosomal release and cytoplasmic delivery of fluorescently labeled TAT-SA complexes with PPAA was shown by the diffuse distribution of fluorescent protein in the cytoplasm. Taken together, these results demonstrate that TAT-SA can be used to direct intracellular delivery of large biotinylated cargo to intracellular compartments and that biotinylated PPAA can direct cytoplasmic delivery where desired. PMID- 15820984 TI - Analysis of the causes and consequences of decreased antibiotic consumption over the last 5 years in Slovenia. AB - OBJECTIVES: Compared with European countries, the use of antibiotics in Slovenia is moderate. In the period 1999-2002 an 18.67% decrease in outpatient antibiotic consumption was noted. The aim of the present study was to analyse this decrease and its consequences. METHODS: The data on outpatient antibiotic consumption were obtained from the Institute of Public Health and Health Insurance Institute of Slovenia and expressed in defined daily doses (DDD)/1000 inhabitant-days. The number of media publications on 'antibiotic drugs' and 'bacterial resistance' during the study period was obtained. In 2000, the prescription of co-amoxiclav and fluoroquinolones was restricted because of a constant increase in the consumption of these drugs. The data on incidence of acute mastoiditis and penicillin resistance among invasive pneumococci were obtained. RESULTS: The total outpatient consumption of antibacterials increased from 15.21 DDD/1000 inhabitant-days in 1996 to 20.08 in 1999, and decreased to 16.97 in 2003. The consumption of restricted antibiotics decreased from 7.29 in 1999 to 5.25 DDD/1000 inhabitant-days in 2003. There was a positive correlation between antibiotic consumption and the number of newspaper articles (r=0.92), and a negative correlation between the number of diagnostic tests and antibiotic consumption (r=-0.73 for the C-reactive protein test and -0.68 for the streptococcal antigen detection test). Reduced antibiotic consumption was paralleled by a decrease in penicillin resistance among invasive pneumococci. No increase in mastoiditis cases was observed in spite of reduced antibiotic consumption. CONCLUSION: Restriction of antibiotic prescription proved to be effective in reducing outpatient antibiotic consumption. The effect was prolonged and affected restricted antibiotics as well as non-restricted drugs. PMID- 15820983 TI - Antimicrobial susceptibility and serotype distribution of Streptococcus pneumoniae causing meningitis in Egypt, 1998-2003. AB - OBJECTIVES: To determine the antimicrobial susceptibility and serotype distribution of 205 isolates of Streptococcus pneumoniae, collected from the CSF of meningitis patients identified between 1998-2003, during sentinel meningitis surveillance in Egypt. METHODS: Antimicrobial susceptibility was evaluated against six antibiotics using disc diffusion and Etest methods. Serotyping was performed by latex agglutination and the Quellung test. RESULTS: Forty-nine percent of all isolates were found to be non-susceptible to penicillin (46% intermediate, MIC range 0.12-1.0 mg/L; 3% resistant, MIC = 2.0 mg/L), and 6% of the isolates were non-susceptible to ceftriaxone (5% intermediate, MIC = 1.0 mg/L; 1.3% resistant, MIC >/= 2 mg/L). Resistance rates for tetracycline and trimethoprim/sulfamethoxazole were high (52 and 59.7%, respectively), but those for erythromycin and chloramphenicol were lower (11 and 9%, respectively). Five serotypes (6B, 1, 19A, 23F and 6A) accounted for 37% of the total isolates. Ten isolates (5%) were non-typeable. Overall, 29 and 42% of serotypes were represented in the 7- and 11-valent conjugate vaccines, respectively. However, vaccine coverage for children <2 years was 38 and 56% for the 7- and 11-valent, respectively. CONCLUSIONS: Resistance to penicillin may be increasing among S. pneumoniae strains causing meningitis in Egypt, and a moderate proportion of these strains are not covered by current pneumococcal conjugate vaccines. In addition to intensifying education efforts about judicious use of antibiotics, laboratory-based surveillance for other forms of invasive pneumococcal disease, especially pneumonia, is needed before decisions can be made regarding the most effective vaccines for control of this disease in Egypt. PMID- 15820985 TI - Exposure of Candida albicans to antifungal agents affects expression of SAP2 and SAP9 secreted proteinase genes. AB - OBJECTIVES: To ascertain the effects of subinhibitory concentrations of several antifungal agents on a virulence factor: secreted proteinase (Sap) activity and expression of SAP genes in Candida albicans. METHODS: Enzyme assays and growth measurements, GFP-SAP2 promoter constructs and fluorescence measurement, transcript profiling and RT-PCR. RESULTS: For seven of eight C. albicans isolates tested, exposure to fluconazole gave an increase in Sap specific activity; for one isolate, resistant to azoles and flucytosine, fluconazole exposure led to a fall in Sap activity. A similar relationship between growth reduction and increased Sap activity was seen with C. albicans cells treated with subinhibitory concentrations of itraconazole, miconazole, flucytosine and caspofungin. Transcript profiling indicated antifungal exposure was associated with increased expression of mRNA from SAP2 and SAP9 genes; this was confirmed for fluconazole and caspofungin exposure by RT-PCR. CONCLUSIONS: Antifungal agents with three different mechanisms of action similarly generate a rise in expression of SAP2 and activity of the secreted Sap2 gene product, a known virulence factor, in most isolates of C. albicans. One isolate of the fungus showed an opposite response. PMID- 15820986 TI - Breeding system, flower visitors and seedling survival of two endangered species of Helianthemum (Cistaceae). AB - BACKGROUND AND AIMS: Helianthemum marifolium and H. caput-felis are two endangered plant species of the western Mediterranean. Several aspects of the reproduction of both species were examined to determine whether their rarity could be related to factors causing reproductive limitation. METHODS: The flowering and fruiting phenology of both species in two non-sympatric island populations (Mallorca, Balearic Islands, western Mediterranean) were compared. Hand-pollination experiments were conducted to determine their fruit and seed production under different pollen sources. The composition of the pollinator assemblage and the effect of temporal variation and sun exposure on reproductive output and seedling survival were also investigated. KEY RESULTS: Flowering periods were longer for H. marifolium than for H. caput-felis in the populations studied. Helianthemum marifolium is mostly an outbreeder, i.e. fruit and seed set was three-fold higher when pollen came from other plants. In H. caput-felis, neither fruit nor seed set was affected by pollination treatments. Flower visitors were more diverse for H. caput-felis than for H. marifolium. In both species, most floral visits were made by hymenopterans. The total number of pollinator visits varied significantly between years, decreasing more than two fold from 2001 to 2002, in both species. In agreement with its outbreeder character, variation in reproductive output between years was also observed in H. marifolium. It showed a 50 % decrease in fruit set in 2002, a pollinator-poor year. Finally, seedling survival increased three- to six-fold from 2001 to 2002. A correlation between seedling size and survival had also been detected. CONCLUSIONS: Reproductive limitations were detected for neither species (i.e. fruit and seed set, pollination service and seedling survival on natural populations). Hence, the increasing rarity of these two species is probably a direct result of the destruction of their habitat. PMID- 15820987 TI - Rice morphogenesis and plant architecture: measurement, specification and the reconstruction of structural development by 3D architectural modelling. AB - BACKGROUND AND AIMS: The morphogenesis and architecture of a rice plant, Oryza sativa, are critical factors in the yield equation, but they are not well studied because of the lack of appropriate tools for 3D measurement. The architecture of rice plants is characterized by a large number of tillers and leaves. The aims of this study were to specify rice plant architecture and to find appropriate functions to represent the 3D growth across all growth stages. METHODS: A japonica type rice, 'Namaga', was grown in pots under outdoor conditions. A 3D digitizer was used to measure the rice plant structure at intervals from the young seedling stage to maturity. The L-system formalism was applied to create '3D virtual rice' plants, incorporating models of phenological development and leaf emergence period as a function of temperature and photoperiod, which were used to determine the timing of tiller emergence. KEY RESULTS: The relationships between the nodal positions and leaf lengths, leaf angles and tiller angles were analysed and used to determine growth functions for the models. The '3D virtual rice' reproduces the structural development of isolated plants and provides a good estimation of the tillering process, and of the accumulation of leaves. CONCLUSIONS: The results indicated that the '3D virtual rice' has a possibility to demonstrate the differences in the structure and development between cultivars and under different environmental conditions. Future work, necessary to reflect both cultivar and environmental effects on the model performance, and to link with physiological models, is proposed in the discussion. PMID- 15820988 TI - Effects of burial in sand and water supply regime on seedling emergence of six species. AB - BACKGROUND AND AIMS: Air seeding has long been regarded as a quick and successful measure for vegetation rehabilitation in China. However, seedling emergence of often-used species including Agriophyllum squarrosum, Artemisia sphaerocephala, Artemisia ordosica, Hedysarum fruticosum, Caragana korshinskii and Medicago sativa is low. Experiments were conducted under controlled conditions to study the effects of sowing depth and water supply on seedling emergence, in order to understand the requirements for increasing seedling emergence. METHODS: Seeds were exposed to different environments of burial and water supply regimes in PVC pots (7 cm in diameter and 11 cm in height) under the same light intensity and alternating temperature regimes in a growth chamber. KEY RESULTS: Seedlings of three species (Agriophyllum squarrosum, Artemisia sphaerocephala, Artemisia ordosica) with relatively light seeds emerged well at a 0.5 cm sowing depth under a 7.5 and 10 mm water supply regime. However, few seedlings of these species emerged when the sowing depth was over 1 cm or when water supply was 5 mm. Seedlings of Caragana korshinskii, Hedysarum fruticosum and Medicago sativa emerged from sowing depths of 0.5-4 cm, 0.5-3 cm, and 0.5-4 cm, respectively, under both 7.5 and 10 mm water supply regimes. Under a 5 mm water supply regime, seedlings of these species also emerged at over 1 cm sowing depth. Seeds of all six species sown on the surface of sand did not germinate, and seedlings did not emerge when they were sown at depths greater than 6 cm. CONCLUSIONS: Based on these experiments, a 0.5 cm sowing depth resulted in the highest seedling emergence and it is concluded that this is the optimal sowing depth for seedling emergence of all six species. PMID- 15820989 TI - Allozyme variation in the endangered insular endemic Castilleja grisea. AB - BACKGROUND AND AIMS: Genetic diversity in Castilleja grisea, an endangered, perennial herb endemic to San Clemente Island, California was investigated. Subsequent to the elimination of goats from the island in 1992, many populations of C. grisea have reappeared and have been increasing in size. METHODS: Nineteen populations were surveyed for their genotype at 19 allozyme loci. KEY RESULTS: At the taxon level, 57.9 % of loci are polymorphic with A(P) = 3.09 and H(E) = 0.137. Populations averaged 33.0 % polymorphic loci with A(P) = 2.43 and H(E) = 0.099. Most variation is found within rather than among populations (G(ST) = 0.128), although differentiation among populations is significant. Genetic identities range from I = 0.960 to I = 1.000 with mean I = 0.990. There is no significant relationship between genetic and geographic distance. Gene flow among populations is Nm = 2.50 based on private alleles and Nm = 1.70 based on F(ST). Outcrossing rates based on fixation indices average t = 1.01, indicating a primarily out-crossed mating system. CONCLUSIONS: The observed genetic variation is moderately high, unusually so for an insular endemic species, suggesting that C. grisea may not have lost substantial genetic variation during 150 years of overgrazing, and indicating that it is unlikely to be endangered by genetic factors. PMID- 15820991 TI - People like us. PMID- 15820992 TI - Lessons in giving vaccines: soft tissue complications of vaccinations. PMID- 15820993 TI - The impact of obesity on health-related quality-of-life in the general adult US population. AB - BACKGROUND: The US Preventive Services Task Force recently recommended screening all adult patients for obesity due in part to the strong association between obesity and numerous chronic diseases. However, how obesity affects health related quality-of-life (HRQL), particularly for persons without any chronic diseases, is less clear. METHODS: The relationship between obesity and HRQL was examined using data from the 2000 Medical Expenditure Panel Survey. Respondents > or =18 years were classified as underweight, normal weight, overweight, class I obesity, and class II obesity based on their BMI. HRQL was measured by the 12 item Short Form physical and mental summary scores (PCS-12 and MCS-12, respectively) and EuroQol EQ-5D index and visual analogue scale (EQ VAS). The impact of obesity on HRQL was examined through multivariate regression, adjusting for sociodemographics and disease status. RESULTS: After adjustment, HRQL decreased with increasing level of obesity. Compared to normal weight respondents, persons with severe obesity had significantly lower scores with scores on the PCS-12, MCS-12, EQ-5D index, and EQ VAS being 4.0, 1.1, 0.073, and 4.8 points lower, respectively. Such decrements of HRQL for severe obesity were similar to the decrements seen for diabetes or hypertension. Persons with moderate obesity or who were overweight also had significantly lower HRQL scores, particularly on the PCS-12 and EQ-5D index. Underweight persons also had lower MCS-12 and EQ VAS scores. CONCLUSIONS: Persons with obesity had significantly lower HRQL than those who were normal weight and such lower scores were seen even for persons without chronic diseases known to be linked to obesity. PMID- 15820994 TI - Evidence-based public health: what does it offer developing countries? AB - The global burden of disease and illness is primarily situated in developing countries. As developing countries have limited resources, it is particularly important to invest in public health and health promotion strategies that are effective. Systematic reviews are central to evidence-based public health and health promotion practice and policy. This paper discusses issues surrounding the relevance of evidence-based public health and systematic reviews to the health of developing countries. It argues that there is a lack of systematic reviews relevant to the health priorities of developing countries; many interventions reviewed can not be implemented in resource-poor situations; and, a limited amount of primary research is conducted in developing countries. The paper further argues that improvements in public health are determined not only by effective health services and interventions, but through an approach that includes other sectors and influences broader structural and systematic barriers to health. Given the social complexity of human development, and the inter sections amongst different development goals, there is no question that gains in developing country public health are unlikely to emerge from systematic reviews alone, but will require decisions about inter-sectoral collaboration and social policy initiatives. Nonetheless, evidence around intervention effectiveness has an important role to play in addressing health priorities in developing countries and resource-poor areas. The public health evidence base urgently needs strengthening, with dedicated effort towards increasing the relevance of primary evidence and systematic reviews. PMID- 15820995 TI - Oral history--professor Alwyn Smith. PMID- 15820996 TI - The Resolve to Stop the Violence Project: transforming an in-house culture of violence through a jail-based programme. AB - BACKGROUND: The usual modes of incarceration have not been found to curb violence significantly, even while in custody. A jail-based programme called the Resolve to Stop the Violence Project (RSVP) was created with the hypothesis that immersing men with a history of serious, recent and often multiple violent crimes in an intensive, multi-modal in-house 'culture' would serve as a possible first step to preventing further violence. METHODS: Two years of incident reports were reviewed for the programme dorm and a regular dorm, both typically serving an average of 56 male inmates of similar composition, for historic and between-dorm comparisons. RESULTS: During the year before RSVP began, there were 24 violent incidents serious enough to have constituted felonies had they occurred in the community (roughly three per month) in the 62-bed dorm. During the first month RSVP was in effect there was one such incident; and for the following 12 months, there were none. During that same year, the control dorm that still followed traditional jail practices had 28 violent incidents. CONCLUSIONS: Correctional efforts may improve with the transformation of subcultures into therapeutic communities that facilitate the practice of prosocial skills over attitudes and mores that engender violence. PMID- 15820997 TI - The Resolve to Stop the Violence Project: reducing violence in the community through a jail-based initiative. AB - BACKGROUND: The usual modes of incarceration have not been found to curb violent crimes significantly. A jail-based programme called the Resolve to Stop the Violence Project (RSVP) was created with the hypothesis that exposing men with a history of serious, recent and often multiple violent crimes to a certain specifiable set of social, cultural and psychological conditions would reduce the frequency and severity of their violent behaviour. METHODS: Court and criminal records for 1 year following release were reviewed for 101 inmates who had spent 8 weeks or more in the programme and for the same number of those who had spent 8 weeks or more in regular custody. RESULTS: Inmates who participated in RSVP had lower rearrest rates for violent crimes (-46.3 percent, p < 0.05) and spent less time in custody (-42.6 percent, p < 0.05). The decline in violent re-arrests increased with greater lengths of stay (-53.1 percent, p < 0.05 for 12 weeks or more; -82.6 percent, p < 0.05 for 16 weeks or more). CONCLUSIONS: Multilevel, comprehensive prevention approaches that: emphasize making available to violent individuals the kinds of tools they need in order to develop non-violent skills and reality-based sources of self-esteem; increase their capacity to experience feelings of empathy and remorse; and provide opportunities to take responsibility and amend the injuries they have inflicted on others and on the whole community, may play an important role in reducing the cycle of violent crime. PMID- 15820998 TI - Impact of statins in microalbuminuric subjects with the metabolic syndrome: a substudy of the PREVEND Intervention Trial. AB - AIMS: Microalbuminuria frequently clusters with the metabolic syndrome and may identify subjects at increased coronary risk. Statin treatment may reduce the incidence of major adverse cardiac events in subjects with the metabolic syndrome, but evidence is limited. We evaluated the impact of pravastatin treatment on the incidence of major adverse cardiac events in microalbuminuric subjects with the metabolic syndrome. METHODS AND RESULTS: This substudy of the PREVEND Intervention Trial (a randomized, placebo-controlled trial with a 2x2 factorial design) included 864 microalbuminuric subjects, who were randomized to fosinopril 20 mg or matching placebo and pravastatin 40 mg or matching placebo (mean follow-up 46 months). The metabolic syndrome was defined according to the NCEP ATPIII-report. Subjects with or without the metabolic syndrome were characterized by a higher age, male sex, and increased albuminuria. The incidence of major adverse cardiac events in subjects with the metabolic syndrome [9.1%; 95% confidence interval (CI) 6.0-13.0%] was increased vs. those without [3.6%; 95% CI 2.3-5.5%; P=0.007). Pravastatin treatment lowered the incidence of major adverse cardiac events in subjects with the metabolic syndrome after adjustment for age and sex (hazard ratio=0.39; 95% CI 0.17-0.89; P=0.025). CONCLUSION: This study supports the use of statins in microalbuminuric subjects with the metabolic syndrome to reduce the incidence of major adverse cardiac events. PMID- 15820999 TI - Exercise testing to stratify risk in aortic stenosis. AB - AIMS: The aims of this study were to assess the accuracy of exercise testing in predicting symptom onset within 12 months in patients with asymptomatic aortic stenosis and to establish the criteria that define a positive test. METHODS AND RESULTS: A total of 125 patients with aortic stenosis [effective orifice area (EOA) 0.9+/-0.2 cm2] were assessed by Specific Activity Scale (SAS) classification, transthoracic echocardiography, and treadmill exercise testing using the modified Bruce protocol. During follow-up, 36 patients (29%) developed spontaneous symptoms within 12 months. Of these, 26 (72%) had had symptoms revealed by exercise testing and 24 (67%) had severe stenosis (EOA < or = 0.8 cm2). Exercise-limiting symptoms were the only independent predictors of outcome at 12 months, and an abnormal blood pressure response or ST segment depression did not improve the accuracy of the exercise test. The positive predictive accuracy for exercise-induced symptoms was 57% in the whole population and 79% for patients aged <70 in SAS Class I. The negative predictive accuracy was 87% in the whole population and 86% in the subgroup. CONCLUSION: A significant proportion of patients with apparently asymptomatic aortic stenosis experience limiting symptoms on treadmill exercise testing. The subsequent development of spontaneous symptoms is strongly related to the severity of stenosis and to limiting symptoms on exercise testing, but less so to an abnormal blood pressure response or ST segment depression. PMID- 15821000 TI - Drug-eluting stents for chronic total occlusions make sense, but it is too early to close the discussion. PMID- 15821001 TI - Utility and safety of diagnostic pericardiocentesis. PMID- 15821002 TI - Cost-effectiveness of on-pump and off-pump bypass surgery. PMID- 15821003 TI - C-reactive protein in the prediction of cardiovascular and overall mortality in middle-aged men: a population-based cohort study. AB - AIMS: Cut-offs for C-reactive protein concentrations have been recommended for risk stratification, but little is known about how these cut-offs predict cardiovascular risk in population-based cohorts. We therefore assessed the association of C-reactive protein levels with cardiovascular mortality in a population-based cohort of 2321 middle-aged men stratified by the presence of cardiovascular disease (CVD) at baseline. METHODS AND RESULTS: C-reactive protein concentrations were categorized according to current recommendations (1 and 3 mg/L). During the 15 year follow-up, 77 men without CVD and 121 men with CVD at baseline died of CVD. In men without CVD at baseline (n=1476), age-adjusted cardiovascular mortality was 4.1-fold higher (95% CI 2.1-8.2) for C-reactive protein levels between 3.0 and 9.9 mg/L at baseline than for C-reactive protein levels <1.0 mg/L. In men with CVD at baseline (n=845), the corresponding age adjusted cardiovascular mortality was 3.3-fold higher (95% CI 2.0-5.3). Adjustment for conventional CVD risk factors attenuated the risk somewhat. Further adjustment for dietary and lifestyle factors and factors related to insulin resistance did not affect the association. Classification of C-reactive protein by tertiles gave qualitatively similar results, but identified twice as many men at high risk. C-reactive protein levels also predicted overall mortality. CONCLUSION: Currently, recommended cut-offs for C-reactive protein levels identify men at risk for cardiovascular and overall death independently of conventional and other risk factors in a population-based sample of middle-aged men with and without CVD at baseline. Lower cut-offs may better identify men at high risk for cardiovascular death, but improvement of current recommendations will require standardization of C-reactive protein assays. PMID- 15821004 TI - Association between hyper- and hypoglycaemia and 2 year all-cause mortality risk in diabetic patients with acute coronary events. AB - AIMS: The study evaluated the associations between glycometabolic parameters at admission and during hospitalization and 2 year all-cause mortality risk in an unselected cohort of consecutive patients with diabetes admitted for unstable angina or non-Q-wave myocardial infarction to a university hospital during 1988 98. METHODS AND RESULTS: A total of 713 consecutive patients with diabetes were included. During 2 years of follow-up, 242 (34%) patients died. All analyses were retrospective using prospectively collected clinical data. The primary study endpoint was 2 year all-cause mortality collected from the Swedish cause-specific mortality register. In unadjusted analyses, high admission blood glucose (highest vs. lowest quartile: hazard ratio (HR) 2.66; 95% confidence interval (CI) 1.83, 3.86) and hypoglycaemia recorded during hospitalization (hypoglycaemia vs. normal: HR 1.77; 95% CI 1.09, 2.86) were both significantly associated with increased 2 year all-cause mortality risk. These associations remained significant after multivariable adjustment. CONCLUSION: In the setting of acute coronary syndromes (ACS) among patients with diabetes, hyperglycaemia on arrival and hypoglycaemia during hospitalization are both independently associated with worse adjusted all-cause 2 year mortality risk. These observations suggest that the avoidance of both hyper- and hypoglycaemia during ACS events may be of similar importance, and glucose modulation remains an important objective to address in future randomized trials. PMID- 15821005 TI - Serum deoxyribonuclease I activity can be used as a sensitive marker for detection of transient myocardial ischaemia induced by percutaneous coronary intervention. AB - AIMS: Cardiac markers such as troponin T (c-TnT) have proven unsuitable for the detection of early and transient myocardial ischaemia. We recently reported that abrupt elevation of serum deoxyribonuclease I (DNase I) activity in the early stage of acute myocardial infarction could be used as a diagnostic marker. To evaluate whether serum DNase I could be used as a marker of early myocardial ischaemia, we investigated alterations in its levels after transient ischaemia induced during percutaneous coronary intervention (PCI). METHODS AND RESULTS: In 24 consecutive patients with stable angina undergoing elective PCI and 12 patients undergoing coronary angiography (CAG), serum samples were tested for DNase I, creatine kinase isoenzyme MB (CK-MB), and c-TnT before, soon after, and 3 and 12-24 h after completion of the procedures. Serum DNase I activity had risen significantly from baseline by 3 h after PCI in 21 of the 24 PCI patients. The mean per cent difference from baseline in serum DNase I activity 3 h after PCI was 35.9+/-37.5%. Even among the 16 PCI patients whose levels of CK-MB and c TnT were within the normal range, 13 showed elevation of serum DNase I activity from baseline after PCI. In the CAG patient group, DNase I activity levels remained unchanged at all times after CAG. CONCLUSION: Elevation of serum DNase I activity can be used as a sensitive marker for detection of transient myocardial ischaemia. PMID- 15821006 TI - Endogenous urocortins reduce vascular tone and renin-aldosterone/endothelin activity in experimental heart failure. AB - AIMS: To investigate the role of the endogenous urocortin peptides in heart failure (HF) through blockade of the corticotropin-releasing factor receptor 2 (CRF-R2). METHODS AND RESULTS: Eight sheep were administered the CRF-R2 antagonist CRF(9-41) (1.5 mg bolus) before (Normal) and after development of pacing-induced HF. Compared with controls, CRF(9-41) in HF significantly increased mean arterial pressure (MAP) (71+/-2 vs. 75+/-2 mmHg, P=0.0024) and calculated total peripheral resistance (CTPR) (33.3+/-5.2 vs. 39.4+/-5.9 mmHg/L/min, P=0.0455). Similar trends were observed in the Normal state (MAP 87+/ 1 vs. 89+/-2 mmHg, P=0.0689; CTPR 21.9+/-2.0 vs. 24.4+/-2.4 mmHg/L/min, P=0.0731). Left atrial pressure was elevated similarly in both states (Normal P=0.0013; HF P=0.0298), whereas cardiac output tended to be reduced (Normal P=0.0614). CRF(9-41) increased plasma urocortin-I (Normal 10.3+/-0.8 vs. 19.8+/ 1.3 pmol/L, P<0.001; HF 14.4+/-0.9 vs. 25.3+/-0.8 pmol/L, P<0.001), renin (Normal 0.34+/-0.06 vs. 0.41+/-0.02 nmol/L/hr, P=0.013; HF 1.14+/-0.29 vs. 1.57+/-0.36 nmol/L/hr, P=0.0326), aldosterone (Normal 370+/-62 vs. 563+/-99 pmol/L, P=0.0813; HF 662+/-141 vs. 1024+/-209 pmol/L, P=0.095), and endothelin-1 (HF 3.18+/-0.18 vs. 4.74+/-1.04 pmol/L, P=0.0087). MAP, CTPR, renin, and endothelin-1 responses to CRF-R2 antagonism were significantly greater in HF than in the Normal state (P=0.049, 0.0427, 0.0311, and 0.0412, respectively). CONCLUSION: These data suggest that the endogenous urocortin peptides contribute to the suppression of vascular tone and renin-angiotensin-aldosterone/endothelin activation in HF and thus, play a protective compensatory role in this disorder. PMID- 15821007 TI - Long-term clinical and echocardiographic results after successful mitral balloon valvotomy and predictors of long-term outcome. AB - AIMS: To assess the long-term outcome of mitral balloon valvotomy (MBV) and identify predictors of restenosis- and event-free survival. METHODS AND RESULTS: We report the immediate and long-term clinical and echocardiographic results in 493 patients, mean age 31+/-11, who underwent successful MBV and were followed-up for 0.5-15 years (median 5+/-3) with clinical and echocardiographic examination. After MBV, mitral valve area increased from 0.84+/-0.2 to 1.83+/-0.53 cm(2) (P<0.0001) as measured by catheter and from 0.92+/-0.17 to 1.96+/-0.29 cm(2) as measured by two-dimensional echo. Restenosis occurred in 86/493 (17.4%) patients and it was less frequent in patients with low echo score. Actuarial freedom from restenosis at 5, 7, 10, and 13 years were 89+/-1, 81+/-2, 68+/-3, and 51+/-6%, respectively, and was significantly higher in patients with low echo score. Event free survival (death, redo MBV, mitral valve replacement, New York Heart Association functional Class III or IV) at 5, 7, 10, and 13 years were 92+/-1, 87+/-2, 80+/-3, and 74+/-3%, respectively, and was significantly higher for patients with low echo score. Cox regression analysis identified mitral echocardiographic score (MES) >8 as predictors of restenosis (P=0.0004) and MES and age as predictors of event-free survival (P=0.0003 and 0.004, respectively). CONCLUSION: MBV has excellent long-term results for selected patients with mitral stenosis. The long-term outcome after this procedure can be predicted from baseline clinical and valvular characteristics. PMID- 15821008 TI - Lifestyle and cardiovascular disease in middle-aged British men: the effect of adjusting for within-person variation. AB - AIMS: To examine the effect that within-person variation has on the estimated risk associations between cigarette smoking, physical inactivity, and increased body mass index (BMI) and the development of cardiovascular disease (CVD) in middle-aged British men. METHODS AND RESULTS: In total, 6452 men aged 40-59 with no prior evidence of CVD were followed for major CVD events (fatal/non-fatal myocardial infarction or stroke) and all-cause mortality over 20 years; lifestyle characteristics were ascertained at regular points throughout the study. A major CVD event within the first 20 years was observed in 1194 men (18.5%). Use of baseline assessments of cigarette smoking and physical activity in analyses resulted in underestimation of the associations between average cumulative exposure to these factors and major CVD risk. After correction for within-person variation, major CVD rates were over four times higher for heavy smokers (> or =40 cigarettes/day) compared with never smokers and three times higher for physically inactive men compared with moderately active men. Major CVD risk increased by 6% for each 1 kg/m(2) increase in usual BMI. If all men had experienced the risk levels of the men who had never regularly smoked cigarettes, were moderately active, and had a BMI of < or =25 kg/m(2) (6% of the population), 66% of the observed major CVD events would have been prevented or postponed (63% before adjustment for within-person variation). Adjustment for a range of other risk factors had little effect on the results. Similar results were obtained for all-cause mortality. CONCLUSION: Failure to take account of within-person variation can lead to underestimation of the importance of lifestyle characteristics in determining CVD risk. Primary prevention through lifestyle modification has a great preventive potential. PMID- 15821009 TI - The efficacy of a 'master switch gene' HIF-1alpha in a porcine model of chronic myocardial ischaemia. AB - AIMS: Therapeutic angiogenesis is a potential new treatment for patients unsuitable for conventional revascularization strategies. We investigated angiogenesis via a 'master switch gene' hypoxia inducible factor (HIF-1alpha). METHODS AND RESULTS: Ameroid occluders were placed around the left circumflex coronary artery of 74 pigs. Three weeks later, pigs were randomized to receive (i) adenovirus encoding HIF-1alpha (Ad2/HIF-1alpha VP-16 10(10) particles); (ii) plasmid DNA encoding HIF-1alpha (pHIF-1alpha NFkappaB 500 microg); (iii) pHIF 1alpha NFkappaB 2500 microg; and (iv) adenoviral control (Ad2/CMV-empty vector 10(10) particles). Twenty injections (50 microL each) were administered epicardially via re-thoracotomy. Three weeks after gene delivery significant (ANOVA P=0.02) changes in myocardial perfusion during stress were seen in the area adjacent to injections. Post hoc testing (Bonferroni) demonstrated that the AdHIF-1alpha group was significantly (P=0.02) different from the Ad2/control. There were also significant (ANOVA P=0.02) differences in resting left ventricular (LV) function. Post hoc (Bonferroni) showed that the AdHIF-1alpha group was significantly different from the Ad2/control (P=0.03). No significant changes in any parameter were seen with plasmid HIF-1alpha. There were no differences in collateralization or capillary growth. CONCLUSION: Ad2/HIF-1alpha increased myocardial perfusion and improved LV function. Plasmid HIF-1alpha was not associated with improvements in any bioactivity endpoints. PMID- 15821011 TI - Prognostic significance of serum cholesterol levels in patients with idiopathic dilated cardiomyopathy. AB - AIMS: Previous studies indicate that low cholesterol levels are associated with adverse prognosis in heart failure patients, because elevated lipoprotein levels may negate bacterial endotoxin load induced by gastrointestinal congestion. METHODS AND RESULTS: We examined the prognostic significance of lipid levels in a cohort of 422 patients with idiopathic dilated cardiomyopathy (iDCM) [50+/-12 years, 342 males, 80 females, left ventricular ejection fraction (LV-EF): 31.6+/ 10.6%]. During 42 months of follow-up, 86 patients (20.3%) died or received a heart transplant. In univariate Cox regression analysis, reduced LV-EF, high New York Heart Association (NYHA) class, and increased LV end-diastolic diameter (LVEDD) were strong risk factors associated with that endpoint, whereas decreased total cholesterol, HDL-cholesterol, and apoprotein I levels were identified as weak risk predictors. After step-wise multivariable analysis, only LVEDD, NYHA class, and LV-EF emerged as parameters independently contributing to the model predicting risk for death or heart transplantation (P<0.05). Cholesterol levels were positively associated with LV-EF and negatively associated with LVEDD (P<0.05). Circulating sCD14 levels, a marker of endotoxin exposure, were related to cholesterol levels (P<0.05) and LV-EF (P<0.05). CONCLUSION: Decreased cholesterol levels do not independently predict adverse prognosis in patients with iDCM. Our findings indicate that low cholesterol levels are dependent on the severity of cardiac disease. PMID- 15821010 TI - Low mannose-binding lectin and increased complement activation correlate to allograft vasculopathy, ischaemia, and rejection after human heart transplantation. AB - AIMS: Transplant-associated coronary artery disease (TxCAD) is a major cause of post-transplant graft failure. The aim of this study was to investigate a possible role of mannose-binding lectin (MBL) deficiency and complement activation in TxCAD. METHODS AND RESULTS: In a prospective study of heart transplant recipients (n=38) with a follow-up of 5.3+/-1.3 years (range: 0.9 6.6), angiographically verified TxCAD (n=6) was correlated to plasma MBL, complement activation, and endothelial activation (soluble E-selectin). MBL deficiency (<100 ng/mL) was detected in 3/6 patients with TxCAD and in 3/32 with non-TxCAD (Kaplan-Meier, P=0.020). Furthermore, one or more acute rejection episodes were observed in 6/6 of the MBL-deficient patients and in 15/32 of the MBL-sufficient patients (chi(2); P=0.016). Complement activation (C4bc) correlated with soluble E-selectin (r=0.36; P=0.027), both being significantly higher in patients with ischaemia detected in the first biopsy (C4bc: 13.4+/-6.1 AU/mL; E-selectin: 96+/-13 ng/mL) than in those without ischaemia (C4bc: 6.3+/ 0.5; E-selectin: 51+/-6; P=0.037 and 0.002). Finally, terminal complement complex correlated closely with mortality (P=0.002). CONCLUSION: Low MBL was related to the development of TxCAD and acute rejection and increased complement activation correlated to histopathologic ischaemia and mortality after heart transplantation. PMID- 15821012 TI - Proteasome inhibition induces TNFR1 shedding from human airway epithelial (NCI H292) cells. AB - The type 1 55-kDa TNF receptor (TNFR1) is an important modulator of lung inflammation. Here, we hypothesized that the proteasome might regulate TNFR1 shedding from human airway epithelial cells. Treatment of NCI-H292 human airway epithelial cells for 2 h with the specific proteasome inhibitor clasto lactacystin beta-lactone induced the shedding of proteolytically cleaved TNFR1 ectodomains. Clasto-lactacystin beta-lactone also induced soluble TNFR1 (sTNFR1) release from the A549 pulmonary epithelial cell line, as well as from primary cultures of human small airway epithelial cells and human umbilical vein endothelial cells. Furthermore, sTNFR1 release induced by clasto-lactacystin beta lactone was not a consequence of apoptosis or the extracellular release of TNFR1 exosome-like vesicles. The clasto-lactacystin beta-lactone-induced increase in TNFR1 shedding was associated with reductions in cell surface receptors and intracytoplasmic TNFR1 stores that were primarily localized to vesicular structures. As expected, the broad-spectrum zinc metalloprotease inhibitor TNF alpha protease inhibitor 2 (TAPI-2) attenuated clasto-lactacystin beta-lactone mediated TNFR1 shedding, which is consistent with its ability to inhibit the zinc metalloprotease-catalyzed cleavage of TNFR1 ectodomains. TAPI-2 also reduced TNFR1 on the cell surface and attenuated the clasto-lactacystin beta-lactone induced reduction of intracytoplasmic TNFR1 vesicles. This suggests that TNFR1 shedding induced by clasto-lactacystin beta-lactone involves the zinc metalloprotease-dependent trafficking of intracytoplasmic TNFR1 vesicles to the cell surface. Together, these data are consistent with the conclusion that proteasomal activity negatively regulates TNFR1 shedding from human airway epithelial cells, thus identifying previously unrecognized roles for the proteasome and zinc metalloproteases in modulating the generation of sTNFRs. PMID- 15821013 TI - Cyclic stretch increases VEGF expression in pulmonary arterial smooth muscle cells via TGF-beta1 and reactive oxygen species: a requirement for NAD(P)H oxidase. AB - Our previous studies have indicated that transforming growth factor (TGF)-beta1 and VEGF expression are increased in the smooth muscle cell (SMC) layer of the pulmonary vessels of lambs with pulmonary hypertension secondary to increased pulmonary blood flow. Furthermore, we found that TGF-beta1 expression increased before VEGF. Because of the increased blood flow in the shunt lambs, the SMC in the pulmonary vessels are exposed to increased levels of the mechanical force, cyclic stretch. Thus, in this study, using primary cultures of pulmonary arterial SMC isolated from pulmonary arteries of 4-wk-old lambs, we investigated the role of cyclic stretch in the apparent coordinated regulation of TGF-beta1 and VEGF. Our results demonstrated that cyclic stretch induced a significant increase in VEGF expression both at the mRNA and protein levels (P < 0.05). The increased VEGF mRNA was preceded by both an increased expression and secretion of TGF-beta1 and an increase in reactive oxygen species (ROS) generation. In addition, a neutralizing antibody against TGF-beta1 abolished the cyclic stretch-dependent increases in both superoxide generation and VEGF expression. Our data also demonstrated that cyclic stretch activated an NAD(P)H oxidase that was TGF-beta1 dependent and that NAD(P)H oxidase inhibitors abolished the cyclic stretch dependent increase in VEGF expression. Therefore, our results indicate that cyclic stretch upregulates VEGF expression via the TGF-beta1-dependent activation of NAD(P)H oxidase and increased generation of ROS. PMID- 15821014 TI - Inhibition of 20-HETE abolishes the myogenic response during NOS antagonism in the ovine fetal pulmonary circulation. AB - Mechanisms that maintain high pulmonary vascular resistance (PVR) and oppose vasodilation in the fetal lung are poorly understood. In fetal lambs, increased pulmonary artery pressure evokes a potent vasoconstriction, suggesting that a myogenic response contributes to high PVR in the fetus. In adult systemic circulations, the arachidonic acid metabolite 20-hydroxyeicosatetraenoic acid (20 HETE) has been shown to modulate the myogenic response, but its role in the fetal lung is unknown. We hypothesized that acute increases in pulmonary artery pressure release 20-HETE, which causes vasoconstriction, or a myogenic response, in the fetal lung. To address this hypothesis, we studied the hemodynamic effects of N-methylsufonyl-12,12-dibromododec-11-enamide (DDMS), a specific inhibitor of 20-HETE production, on the pulmonary vasoconstriction caused by acute compression of the ductus arteriosus (DA) in chronically prepared fetal sheep. An inflatable vascular occluder around the DA was used to increase pulmonary artery pressure under three study conditions: control, after pretreatment with nitro-L-arginine (L-NA; to inhibit shear-stress vasodilation), and after combined treatment with both L-NA and a specific 20-HETE inhibitor, DDMS. We found that DA compression after L-NA treatment increased PVR by 44 +/- 12%. Although intrapulmonary DDMS infusion did not affect basal PVR, DDMS completely abolished the vasoconstrictor response to DA compression in the presence of L-NA (44 +/- 12% vs. 2 +/- 4% change in PVR, L-NA vs. L-NA + DDMS, P < 0.05). We conclude that 20-HETE mediates the myogenic response in the fetal pulmonary circulation and speculate that pharmacological inhibition of 20-HETE might have a therapeutic role in neonatal conditions characterized by pulmonary hypertension. PMID- 15821015 TI - PKC-dependent, burn-induced adherens junction reorganization and barrier dysfunction in pulmonary microvascular endothelial cells. AB - Rat lung microvascular endothelial cell monolayers were exposed to donor plasma from burned rats (25% total body surface area) at 1:3 dilution for 30 min. Immunofluorescence analysis revealed that concomitant with gap formation alterations were seen in the adherens junction (AJ) proteins beta-catenin and vascular endothelial-cadherin. Both of these components were shown to exist in a smooth, uniform arrangement at the cell periphery in untreated cells. However, upon exposure to burn plasma, this uniformity was lost, and the AJ proteins showed a disrupted, zipper-like pattern at the cells' edge. In addition, these proteins were absent from areas of gap formation between the cells, and an increase in punctate staining throughout the cells suggests they were internalized in response to burn plasma. Measurements of both transendothelial electrical resistance and FITC-albumin flux across the cell monolayer were used to assess barrier integrity. Our study found that exposure to burn plasma rapidly caused the electrical resistance across confluent monolayers to decrease and albumin flux to increase, phenomena associated with barrier dysfunction. Furthermore, all the above responses to burn plasma were attenuated when cells were pretreated with the PKC inhibitor bisindolylmaleimide, suggesting that PKC is required for burn-induced pulmonary endothelial dysfunction. PMID- 15821016 TI - Hyperoxia enhances brain-derived neurotrophic factor and tyrosine kinase B receptor expression in peribronchial smooth muscle of neonatal rats. AB - Airway hyperreactivity is one of the hallmarks of hyperoxic lung injury in early life. As neurotrophins such as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are potent mediators of neuronal plasticity, we hypothesized that neurotrophin levels in the pulmonary system may be disturbed by hyperoxic exposure. We therefore evaluated the effects of hyperoxia on the expression of BDNF, NGF, and their corresponding high-affinity receptors, TrkB and TrkA, respectively, in the lung of rat pups. Five-day-old Sprague-Dawley rat pups were randomized to hyperoxic or control groups and then continuously exposed to hyperoxia (>95% oxygen) or normoxia over 7 days. At both mRNA and protein levels, BDNF was detected in lung but not in trachea; its level was substantially enhanced in lungs from the hyperoxia-exposed rat pups. Distribution of BDNF mRNA by in situ hybridization indicates that peribronchial smooth muscle was the major source of increased BDNF production in response to hyperoxic exposure. Interestingly, hyperoxia-induced elevation of BDNF was not accompanied by any changes of NGF levels in lung. Furthermore, hyperoxic exposure increased the expression of TrkB in peribronchial smooth muscle but had no effect on the distribution of the specific NGF receptor TrkA. These findings indicate that hyperoxic stress not only upregulates BDNF at mRNA and protein levels but also enhances TrkB within peribronchial smooth muscle. However, there was no corresponding effect on NGF and TrkA receptors. We speculate that the increased level of BDNF may contribute to hyperoxia-induced airway hyperresponsiveness in early postnatal life. PMID- 15821017 TI - Deletion of the eNOS gene has a greater impact on the pulmonary circulation of male than female mice. AB - Nitric oxide is involved in development and postnatal adaptation of the pulmonary circulation. This study aimed to determine whether genetic deletion of nitric oxide synthase (NOS) would lead to maldevelopment of the pulmonary arteries in fetal life, compromise adaptation to extrauterine life, and be associated with a pulmonary hypertensive phenotype in adult life and if any abnormalities were detected, were they sex dependent. Morphometric analyses were made on lung tissue from male and female fetal, newborn, 14-day-old, and adult endothelial NOS deficient (eNOS-/-) or inducible NOS-deficient (iNOS-/-) and wild-type mice. Hemodynamic studies were carried out on adult mice with deletion of either eNOS or iNOS genes. We found that in eNOS-/- mice, lung development was normal in fetal, newborn, and adult lungs. Pulmonary arterial muscularity was greater than normal in both male and female eNOS-/- during fetal life and at birth, but the abnormality persisted only in male mice. Right ventricular hypertrophy was present in 14-day-old and adult male eNOS-/- but not in female mice. Adult male eNOS-/- mice had higher mean right ventricular and systemic pressures than female eNOS-/- mice (P < 0.05). Thus deletion of the eNOS gene was associated with structural evidence of pulmonary hypertension in both sexes during fetal life, but pulmonary hypertension persisted only in the male. In neither sex did iNOS or neuronal NOS appear to compensate for the eNOS deletion. Adult iNOS-/- mice did not have structural or hemodynamic evidence of pulmonary hypertension. Possible compensatory mechanisms are discussed. PMID- 15821018 TI - CD38/cyclic ADP-ribose signaling: role in the regulation of calcium homeostasis in airway smooth muscle. AB - The contractility of airway smooth muscle cells is dependent on dynamic changes in the concentration of intracellular calcium. Signaling molecules such as inositol 1,4,5-trisphosphate and cyclic ADP-ribose play pivotal roles in the control of intracellular calcium concentration. Alterations in the processes involved in the regulation of intracellular calcium concentration contribute to the pathogenesis of airway diseases such as asthma. Recent studies have identified cyclic ADP-ribose as a calcium-mobilizing second messenger in airway smooth muscle cells, and modulation of the pathway involved in its metabolism results in altered calcium homeostasis and may contribute to airway hyperresponsiveness. In this review, we describe the basic mechanisms underlying the dynamics of calcium regulation and the role of CD38/cADPR, a novel pathway, in the context of airway smooth muscle function and its contribution to airway diseases such as asthma. PMID- 15821019 TI - PGE2 and PAR-1 in pulmonary fibrosis: a case of biting the hand that feeds you? PMID- 15821020 TI - IGF-I: mediator of fibrosis or carcinogenesis? PMID- 15821021 TI - Isoproterenol induces actin depolymerization in human airway smooth muscle cells via activation of an Src kinase and GS. AB - In a previous study, we showed that isoproterenol induced actin depolymerization in human airway smooth muscle cells by both protein kinase A (PKA)-dependent and independent signaling pathways. We now investigate the signaling pathway of PKA independent actin depolymerization induced by isoproterenol in these cells. Cells were briefly exposed to isoproterenol or PGE(1) in the presence and absence of specific inhibitors of Src-family tyrosine kinases, phosphatidylinositol-3-kinase (PI3 kinase), or MAP kinase, and actin depolymerization was measured by concomitant staining of filamentous actin with FITC-phalloidin and globular actin with Texas red DNase I. Isoproterenol, cholera toxin, and PGE(1) induced actin depolymerization, indicated by a decrease in the intensity of filamentous/globular fluorescent staining. Pretreatment with the Src kinase inhibitors 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyriimidine (PP2) or geldanamycin or the PKA inhibitor Rp-cAMPS only partly inhibited isoproterenol or PGE(1)-induced actin depolymerization. In contrast, PP2 and geldanamycin did not inhibit forskolin-induced actin depolymerization, and AG-213 (an EGF receptor tyrosine kinase inhibitor) did not inhibit isoproterenol- or PGE(1)-induced actin depolymerization. PI3 kinase or MAP kinase inhibition did not inhibit isoproterenol-induced actin depolymerization. Moreover, isoproterenol but not forskolin induced tyrosine phosphorylation of an Src family member at position 416. These results further confirm that both PKA-dependent and PKA-independent pathways mediate actin depolymerization in human airway smooth muscle cells and that the PKA-independent pathway by which isoproterenol induces actin depolymerization in human airway smooth muscle cells involves Src protein tyrosine kinases and the G(s) protein. PMID- 15821022 TI - Gene expression of ADP-glucose pyrophosphorylase and starch contents in rice cultured cells are cooperatively regulated by sucrose and ABA. AB - Six cDNA clones encoding two small subunits and four large subunits of ADP glucose pyrophosphorylase (AGPase) were mined from the database of rice full length cDNAs, cloned and subsequently named: OsAPS1, OsAPS2, OsAPL1, OsAPL2, OsAPL3 and OsAPL4. Expression patterns of the six genes were examined by Northern blot analysis with gene-specific probes. OsAPL3 was predominantly expressed in the middle phases of seed development, and OsAPS1, OsAPL1 and OsAPL2 were expressed later in seed development. OsAPS2 and OsAPL4 were constitutively expressed and these isoforms were coordinated with starch accumulation in the developing rice seed. In order to clarify the effect of sugars and plant hormones on AGPase gene expression more precisely, a rice cell culture system was used. OsAPL3 transcript significantly accumulated in response to increased levels of sucrose and abscisic acid (ABA) concentration in the medium; however, the transcripts of other AGPase genes did not show significant accumulation. Under identical conditions, starch contents in the cultured cells also increased. Interestingly, ABA alone did not affect the gene expression of OsAPL3 and starch content. Collectively, these results indicated that the expression level of OsAPL3 and starch content in the cultured cells were cooperatively controlled by alterations in the concentration of both sucrose and ABA. PMID- 15821023 TI - Involvement of alpha-amylase I-1 in starch degradation in rice chloroplasts. AB - To determine the role of alpha-amylase isoform I-1 in the degradation of starch in rice leaf chloroplasts, we generated a series of transgenic rice plants with suppressed expression or overexpression of alpha-amylase I-1. In the lines with suppressed expression of alpha-amylase I-1 at both the mRNA and protein levels, seed germination and seedling growth were markedly delayed in comparison with those in the wild-type plants. However, the growth retardation was overcome by supplementation of sugars. Interestingly, a significant increase of starch accumulation in the young leaf tissues was observed under a sugar-supplemented condition. In contrast, the starch content of leaves was reduced in the plants overexpressing alpha-amylase I-1. In immunocytochemical analysis with specific anti-alpha-amylase I-1 antiserum, immuno-gold particles deposited in the chloroplasts and extracellular space in young leaf cells. We further examined the expression and targeting of alpha-amylase I-1 fused with the green fluorescent protein in re-differentiated green cells, and showed that the fluorescence of the expressed fusion protein co-localized with the chlorophyll autofluorescence in the transgenic cells. In addition, mature protein species of alpha-amylase I-1 bearing an oligosaccharide side chain were detected in the isolated chloroplasts. Based on these results, we concluded that alpha-amylase I-1 targets the chloroplasts through the endoplasmic reticulum-Golgi system and plays a significant role in the starch degradation in rice leaves. PMID- 15821024 TI - Circadian G2 arrest as related to circadian gating of cell population growth in Euglena. AB - Cell population growth is gated to occur in particular circadian phases, which has been known for over four decades in various organisms including cyanobacteria and human. However, little is known as to which cell cycle phases from G1 to M are primarily regulated by the circadian rhythm or when in a circadian cycle this primary regulation takes place. We report here that in the flagellate alga Euglena gracilis grown photoautotrophically, the circadian rhythm primarily prevented developmentally matured G2 cells from progressing to mitosis, such that cell population growth occurred only during subjective night. In addition, we found that the circadian rhythm also arrests G1-to-S and S-to-G2 transitions at particular circadian phases. PMID- 15821025 TI - A link between citrate and proton release by proteoid roots of white lupin (Lupinus albus L.) grown under phosphorus-deficient conditions? AB - White lupin (Lupinus albus L.) is able to acclimate to phosphorus deficiency by forming proteoid roots that release a large amount of citric acid, resulting in the mobilization of sparingly soluble soil phosphate in the rhizosphere. The mechanisms responsible for the release of organic acids have not been fully elucidated. In this study, we focused on the link between citrate and malate release and the release of H+ and other inorganic ions by proteoid roots of white lupin. The release of citrate was closely correlated with the release of H+, K+, Na+ and Mg2+, but not with that of Ca2+. The stoichiometric relationships between citrate release and the release of H+, K+, Na+ and Mg2+ were 1 : 1.3, 1 : 2.1, 1 : 1.5 and 1 : 0.47, respectively. Similar correlations were found between exudation of malate and cations. During 30 min incubation, fusicoccin addition stimulated H+ and malate release, but not citrate release. A concomitant stimulation of H+, malate and citrate release was measured after 60 min incubation. Vanadate inhibited the release of H+ and malate, but not that of citrate. Anthracene-9-carboxylic acid, an anion channel blocker, caused a concomitant decrease in release of citrate, malate and H+. We conclude that for export of citrate across the plasma membrane of proteoid root cells, H+ release is not strictly related to citrate release. Other cations such as K+ and Na+ can also serve as counterions for citrate release. In contrast, malate release shows a strong H+ release dependency. PMID- 15821026 TI - JTP-27536 [(+)-1,3-dihydroxy-2-hydroxymethylpropyl-2-ammonium 2-[(R)-3-cyclo hexyl-1-phenylpropyl]-1,3-dioxo-2,3-dihydro-1H-isoindole-5-carboxylate monohydrate], a novel inhibitor of immunoglobulins and interleukin-5 with anti inflammatory properties in mouse allergic dermatitis model. AB - We report a novel synthetic compound JTP-27536 [(+)-1,3-dihydroxy-2 hydroxymethylpropyl-2-ammonium 2-[(R)-3-cyclohexyl-1-phenylpropyl]-1,3-dioxo-2,3 dihydro-1H-isoindole-5-carboxylate monohydrate] as an inhibitor of immunoglobulins (Igs) and interleukin (IL)-5 production in vitro and in vivo. JTP 27536 inhibited IgE production in mouse and human B cells with IC50 values of 2.5 and 2.1 microM, respectively, and the inhibition was stronger than that on IgG1 and IgM production (IC50 > 10 microM). JTP-27536 also inhibited IL-5 production in mouse splenocytes and human peripheral blood mononuclear cells with IC50 values of 3.3 and 1.3 microM, respectively, without affecting mouse interferon (IFN)-gamma, IL-2, IL-4, IL-10, or human IL-4 production. In contrast, prednisolone not only inhibited mouse IgE production but also mouse IFN-gamma, IL 2, IL-4, and IL-10 and human IL-4 and IL-5 production in vitro. The effect of suplatast tosilate, a Th2 cytokine inhibitor, on antibody and cytokine production was less potent than that of JTP-27536. In vivo animal experiments using dinitrophenylated ascaris-sensitized mice and 2,4,6-trinitro-1-chrolobenzene induced chronic dermatitis mice showed that JTP-27536 was more potent than suplatast tosilate and comparable with prednisolone in inhibiting ear swelling, antigen-specific IgE and IL-5 production, and cell infiltrations into the inflamed tissue. These results indicate that JTP-27536 is an inhibitor of Igs, in particular IgE, and of IL-5, which has antiallergic properties in mouse dermatitis model, and suggest that an inhibitor of Igs and IL-5 like JTP-27536 may be useful as a drug for the treatment of allergic diseases. PMID- 15821027 TI - G protein-dependent pharmacology of histamine H3 receptor ligands: evidence for heterogeneous active state receptor conformations. AB - Previously reported pharmacological studies using the imidazole-containing histamine H3 receptor ligands GT-2331 (Cipralisant) and proxyfan resulted in a range of classifications (antagonist, agonist, and protean) for these compounds. We examined the role that the signaling system, with particular emphasis on the type of G protein, had on the pharmacology observed for H3 ligands. Ligands were assessed using assays measuring neurotransmitter release, cAMP, and guanosine 5' O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding. Whereas clobenpropit and ciproxifan were consistently antagonists, GT-2331, proxyfan, and imetit exhibited differential activity. Although GT-2331 and proxyfan exhibited little agonist activity in neurotransmitter release assays, both demonstrated full agonism relative to (R)-alpha-methylhistamine in cAMP assays. In [35S]GTPgammaS binding assays, GT-2331 and proxyfan demonstrated partial agonism. Imetit showed full agonism in most assays, but it was slightly less efficacious in a neurotransmitter release assay and in [35S]GTPgammaS binding at the human H3 receptor. To further examine these ligands, we coexpressed G alpha16 or chimeric G alpha q/i5 in human embryonic kidney cells expressing the human H3 receptor and assayed intracellular calcium and cAMP levels. GT-2331, proxyfan, and imetit demonstrated full agonism in all assays of cAMP activity. However, in cells expressing G alpha16, they exhibited minimal agonism in calcium mobilization assays, whereas imetit showed partial agonism. When G alpha q/i5 was used, the activity of both GT-2331 and proxyfan increased, whereas imetit became a full agonist. These results demonstrate that GT-2331 and proxyfan's differential pharmacology at the H3 receptor depends on the type of G protein used and provide indirect evidence for differential ligand-bound active states that mediate signaling by the H3 receptor. PMID- 15821028 TI - Electrical addressing of confined quantum systems for quasiclassical computation and finite state logic machines. AB - Conduction spectroscopy measures the current I through a nanosystem as a function of the voltage V between two electrodes. The differential conductance, dI/dV, has peaks that can be assigned to resonance conditions with different electronic levels of the system. Between these increments, the current has roughly constant plateaus. We discuss how measurements of the current vs. voltage can be used to perform Boolean operations and hence construct finite state logic machines and combinational circuits. The inputs to the device are the source-drain voltage, including its sign, and a gate voltage applied in a manner analogous to optical Stark spectroscopy. As simple examples, we describe a two-state set-reset machine (a machine whose output depends on the input and also on its present state) and a full adder circuit (a circuit that requires three inputs and provides two outputs). PMID- 15821029 TI - Linker histone variants control chromatin dynamics during early embryogenesis. AB - Complex transitions in chromatin structure produce changes in genome function during development in metazoa. Linker histones, the last component of nucleosomes to be assembled into chromatin, comprise considerably divergent subtypes as compared with core histones. In all metazoa studied, their composition changes dramatically during early embryogenesis concomitant with zygotic gene activation, leading to distinct functional changes that are still poorly understood. Here, we show that early embryonic linker histone B4, which is maternally expressed, is functionally different from somatic histone H1 in influencing chromatin structure and dynamics. We developed a chromatin assembly system with nucleosome assembly protein-1 as a linker histone chaperone. This assay system revealed that maternal histone B4 allows chromatin to be remodeled by ATP-dependent chromatin remodeling factor, whereas somatic histone H1 prevents this remodeling. Structural analysis shows that histone B4 does not significantly restrict the accessibility of linker DNA. These findings define the functional significance of developmental changes in linker histone variants. We propose a model that holds that maternally expressed linker histones are key molecules specifying nuclear dynamics with respect to embryonic totipotency. PMID- 15821030 TI - The nucleotide switch in Cdc42 modulates coupling between the GTPase-binding and allosteric equilibria of Wiskott-Aldrich syndrome protein. AB - The GTP/GDP nucleotide switch in Ras superfamily GTPases generally involves differential affinity toward downstream effectors, with the GTP-bound state having a higher affinity for effector than the GDP-bound state. We have developed a quantitative model of allosteric regulation of the Wiskott-Aldrich syndrome protein (WASP) by the Rho GTPase Cdc42 to better understand how GTPase binding is coupled to effector activation. The model accurately predicts WASP affinity for Cdc42, activity toward Arp2/3 complex, and activation by Cdc42 as functions of a two-state allosteric equilibrium in WASP. The ratio of GTPase affinities for the inactive and active states of WASP is appreciably larger for Cdc42-GTP than for Cdc42-GDP. The greater ability to distinguish between the two states of WASP makes Cdc42-GTP a full WASP agonist, whereas Cdc42-GDP is only a partial agonist. Thus, the nucleotide switch controls not only the affinity of Cdc42 for its effector but also the efficiency of coupling between the Cdc42-binding and allosteric equilibria in WASP. This effect can ensure high fidelity and specificity in Cdc42 signaling in crowded membrane environments. PMID- 15821032 TI - Daily administration of interleukin-18 causes myocardial dysfunction in healthy mice. AB - Although increased levels of circulating interleukin (IL)-18 have been demonstrated in patients with cardiovascular diseases, the functional consequences of chronically increased circulating IL-18 with respect to myocardial function have not been defined. Thus we aimed to examine the effects of chronic IL-18 exposure on left ventricular (LV) function in healthy mice. Moreover, to clarify whether IL-18 has direct effects on the cardiomyocyte, we examined effects of IL-18 on cardiomyocytes in vitro. After 7 days of daily intraperitoneal injections of 0.5 microg IL-18 in healthy mice, a 40% (P < 0.05) reduction in the LV maximal positive derivative, a 25% (P < 0.05) reduction in the LV maximal rate of pressure decay, and a 2.8-fold (P < 0.001) increase in the LV end-diastolic pressure were measured, consistent with myocardial dysfunction. Furthermore, we measured a 75% (P < 0.05) reduction in beta-adrenergic responsiveness to isoproterenol. IL-18 induced myocardial hypertrophy, and there was a 2.9-fold increase (P < 0.05) in atrial natriuretic peptide mRNA expression in the LV myocardium. In vitro examinations of isolated adult rat cardiomyocytes being stimulated with IL-18 (0.1 microg/ml) exhibited an increase in peak Ca2+ transients (P < 0.05) and in diastolic Ca2+ concentrations (P < 0.05). In conclusion, this study shows that daily administration of IL-18 in healthy mice causes LV myocardial dysfunction and blunted beta-adrenergic responsiveness to isoproterenol. A direct effect of IL-18 on the cardiomyocyte in vitro was demonstrated, suggesting that IL-18 reduces the responsiveness of the myofilaments to Ca2+. Finally, induction of myocardial hypertrophy by IL-18 indicates a role for this cytokine in myocardial remodeling. PMID- 15821033 TI - S-nitroso-albumin carries a thiol-labile pool of nitric oxide, which causes venodilation in the rat. AB - It is now established that S-nitroso-albumin (SNO-albumin) circulates at low nanomolar concentrations under physiological conditions, but concentrations may increase to micromolar levels during disease states (e.g., cirrhosis or endotoxemia). This study tested the hypothesis that high concentrations of SNO albumin observed in some diseases modulate vascular function and that it acts as a stable reservoir of nitric oxide (NO), releasing this molecule when the concentrations of low-molecular-weight thiols are increased. SNO-albumin was infused into rats to increase the plasma concentration from <50 nmol/l to approximately 4 micromol/l. This caused a 29 +/- 6% drop in blood pressure, 20 +/ 4% decrease in aortic blood flow, and a 25 +/- 14% reduction of renal blood flow within 10 min. These observations were in striking contrast to those of an infused arterial vasodilator (hydralazine), which increased aortic blood flow, and suggested that SNO-albumin acts primarily as a venodilator in vivo. This was confirmed by the observations that glyceryl trinitrate (a venodilator) led to similar hemodynamic changes and that the hemodynamic effects of SNO-albumin are reversed by infusion of colloid. Infusion of N-acetylcysteine into animals with artificially elevated plasma SNO-albumin concentrations led to the rapid decomposition of SNO-albumin in vivo and reproduced the hemodynamic effects of SNO-albumin infusion. These data demonstrate that SNO-albumin acts primarily as a venodilator in vivo and represents a stable reservoir of NO that can release NO when the concentrations of low-molecular-weight thiols are elevated. PMID- 15821034 TI - Structural differences in two biochemically defined populations of cardiac mitochondria. AB - To determine whether there are structural differences in two topologically separated, biochemically defined mitochondrial populations in rat heart myocytes, the interior of these organelles was examined by high-resolution scanning electron microscopy. On the basis of a count of 159 in situ subsarcolemmal mitochondria (SSM, i.e., those that directly abut the sarcolemma), these organelles possess mainly lamelliform cristae (77%), whereas the cristae in in situ interfibrillar mitochondria (IFM, i.e., those situated between the myofibrils, n = 300) are mainly tubular (55%) or a mixture of tubular and lamelliform (24%). Isolated SSM (n = 374), similar to their in situ counterparts, have predominantly lamelliform cristae (75%). The proportions of crista types in isolated IFM (n = 337) have been altered, with only 20% of these organelles retaining exclusively tubular cristae, whereas 58% are mixed; of the latter, lamelliform cristae predominate. This finding suggests that, in contrast to SSM, the cristae in IFM are structurally plastic, changing during isolation. These observations on >1,000 organelles provide the first quantitative morphological evidence for definitive differences between the two populations of cardiac mitochondria. PMID- 15821035 TI - Regulation of cardiac malonyl-CoA content and fatty acid oxidation during increased cardiac power. AB - Myocardial fatty acid oxidation is regulated by carnitine palmitoyltransferase I (CPT I), which is inhibited by malonyl-CoA. Increased cardiac power causes a fall in malonyl-CoA content and accelerated fatty acid oxidation; however, the mechanism for the decrease in malonyl-CoA is unclear. Malonyl-CoA is formed by acetyl-CoA carboxylase (ACC) and degraded by malonyl-CoA decarboxylase (MCD); thus a fall in malonyl-CoA could be due to activation of MCD, inhibition of ACC, or both. This study assessed the effects of increased cardiac power on malonyl CoA content and ACC and MCD activities. Anesthetized pigs were studied under control conditions and during increased cardiac power in response to dobutamine infusion and aortic constriction alone, under hyperglycemic conditions, or with the CPT I inhibitor oxfenicine. An increase in cardiac power was accompanied by increased myocardial O(2) consumption, decreased malonyl-CoA concentration, and increased fatty acid oxidation. There were no differences among groups in activity of ACC or AMP-activated protein kinase (AMPK), which physiologically inhibits ACC. There also were no differences in V(max) or K(m) of MCD. Previous studies have demonstrated that AMPK can be inhibited by protein kinase B (PKB); however, PKB was activated by dobutamine and the elevated insulin that accompanied hyperglycemia, but there was no effect on AMPK activity. In conclusion, the fall in malonyl-CoA and increase in fatty acid oxidation that occur with increased cardiac work were not due to inhibition of ACC or activation of MCD, suggesting alternative regulatory mechanisms for the work-induced decrease in malonyl-CoA concentration. PMID- 15821036 TI - Diverse phenotypes of outward currents in cells that have survived in the 5-day infarcted heart. AB - We have shown reduced density and altered kinetics in slowly activating K+ currents (I(Ks)) in epicardial border zone (EBZ) cells (IZs) of the 5-day infarcted canine heart (Jiang M, Cabo, C, Yao J-A, Boyden PA, and Tseng G-N. Cardiovasc Res 48: 34-43, 2000). beta-Adrenergic stimulation with isoproterenol increases I(Ks) in normal cells (NZs). In this study, we used a voltage-clamp protocol with an external solution to isolate I(Ks) from contaminating currents to determine the effects of 1 muM isoproterenol on I(Ks) in IZs and NZs. Under our recording conditions, 10 microM azimilide-sensitive currents were stimulated with isoproterenol to compare responsiveness of I(Ks) to isoproterenol in the two cell groups. I(Ks) tail density was reduced 67% in IZs (group I, n = 26) compared with NZs (n = 24, P < 0.05). Isoproterenol-stimulated azimilide-sensitive tail currents were increased 1.72 +/- 0.2-fold in NZs and 2.2 +/- 0.3-fold in IZs (P > 0.05). In 33% of IZs (group II, n = 13), native currents showed no tail currents; however, isoproterenol-stimulated azimilide-sensitive currents were voltage dependent, fast activating, and large in amplitude compared with group I IZs, similar to "lone" KCNQ1 currents. Using short clamp pulses, we also found an increase in sustained currents sensitive to tetraethylammonium chloride (TEA) and no change in C-9356-sensitive currents in IZs with little or no transient outward current. In some IZs where I(Ks) is downregulated, the effect of isoproterenol on I(Ks) was similar to that on I(Ks) in NZs. In others, the existence of lone KCNQ1 type currents, which are sensitive to beta-adrenergic stimulation, is consistent with our findings of an increased KCNQ1-to-KCNE1 mRNA ratio (Jiang et al.). Accompanying altered I(Ks) in IZs are an enhanced TEA-sensitive current and a normal C-9356-sensitive current. PMID- 15821038 TI - Attenuation of extracellular ATP response in cardiomyocytes isolated from hearts subjected to ischemia-reperfusion. AB - Extracellular ATP is known to augment cardiac contractility by increasing intracellular Ca2+ concentration ([Ca2+]i) in cardiomyocytes; however, the status of ATP-mediated Ca2+ mobilization in hearts undergoing ischemia-reperfusion (I/R) has not been examined previously. In this study, therefore, isolated rat hearts were subjected to 10-30 min of global ischemia and 30 min of reperfusion, and the effect of extracellular ATP on [Ca2+]i was measured in purified cardiomyocytes by fura-2 microfluorometry. Reperfusion for 30 min of 20-min ischemic hearts, unlike 10-min ischemic hearts, revealed a partial depression in cardiac function and ATP induced increase in [Ca2+]i; no changes in basal [Ca2+]i were evident in 10- or 20-min I/R preparations. On the other hand, reperfusion of 30-min ischemic hearts for 5, 15, or 30 min showed a marked depression in both cardiac function and ATP induced increase in [Ca2+]i and a dramatic increase in basal [Ca2+]i. The positive inotropic effect of extracellular ATP was attenuated, and the maximal binding characteristics of 35S-labeled adenosine 5'-[gamma-thio]triphosphate with crude membranes from hearts undergoing I/R was decreased. ATP-induced increase in [Ca2+]i in cardiomyocytes was depressed by verapamil and Cibacron Blue in both control and I/R hearts; however, this response in I/R hearts, unlike control hearts, was not affected by ryanodine. I/R-induced alterations in cardiac function and ATP-induced increase in [Ca2+]i were attenuated by treatment with an antioxidant mixture and by ischemic preconditioning. The observed changes due to I/R were simulated in hearts perfused with H2O2. The results suggest an impairment of extracellular ATP-induced Ca2+ mobilization in I/R hearts, and this defect appears to be mediated through oxidative stress. PMID- 15821037 TI - Hemodynamic profile, responsiveness to anandamide, and baroreflex sensitivity of mice lacking fatty acid amide hydrolase. AB - The endocannabinoid anandamide exerts neurobehavioral, cardiovascular, and immune regulatory effects through cannabinoid receptors (CB). Fatty acid amide hydrolase (FAAH) is an enzyme responsible for the in vivo degradation of anandamide. Recent experimental studies have suggested that targeting the endocannabinergic system by FAAH inhibitors is a promising novel approach for the treatment of anxiety, inflammation, and hypertension. In this study, we compared the cardiac performance of FAAH knockout (FAAH-/-) mice and their wild-type (FAAH+/+) littermates and analyzed the hemodynamic effects of anandamide using the Millar pressure-volume conductance catheter system. Baseline cardiovascular parameters, systolic and diastolic function at different preloads, and baroreflex sensitivity were similar in FAAH-/- and FAAH+/+ mice. FAAH-/- mice displayed increased sensitivity to anandamide-induced, CB1-mediated hypotension and decreased cardiac contractility compared with FAAH(+/+) littermates. In contrast, the hypotensive potency of synthetic CB1 agonist HU-210 and the level of expression of myocardial CB1 were similar in the two strains. The myocardial levels of anandamide and oleoylethanolamide, but not 2-arachidonylglycerol, were increased in FAAH-/- mice compared with FAAH+/+ mice. These results indicate that mice lacking FAAH have a normal hemodynamic profile, and their increased responsiveness to anandamide induced hypotension and cardiodepression is due to the decreased degradation of anandamide rather than an increase in target organ sensitivity to CB1 agonists. PMID- 15821039 TI - Antioxidant mechanism of heme oxygenase-1 involves an increase in superoxide dismutase and catalase in experimental diabetes. AB - Increased heme oxygenase (HO)-1 activity attenuates endothelial cell apoptosis and decreases superoxide anion (O2-) formation in experimental diabetes by unknown mechanisms. We examined the effect of HO-1 protein and HO activity on extracellular SOD (EC-SOD), catalase, O2-, inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS) levels and vascular responses to ACh in control and diabetic rats. Vascular EC-SOD and plasma catalase activities were significantly reduced in diabetic compared with nondiabetic rats (P < 0.05). Upregulation of HO-1 expression by intermittent administration of cobalt protoporphyrin, an inducer of HO-1 protein and activity, resulted in a robust increase in EC-SOD but no significant change in Cu-Zn-SOD. Administration of tin mesoporphyrin, an inhibitor of HO-1 activity, decreased EC SOD protein. Increased HO-1 activity in diabetic rats was associated with a decrease in iNOS but increases in eNOS and plasma catalase activity. On the other hand, aortic ring segments from diabetic rats exhibited a significant reduction in vascular relaxation to ACh, which was reversed with cobalt protoporphyrin treatment. These data demonstrate that an increase in HO-1 protein and activity, i.e., CO and bilirubin production, in diabetic rats brings about a robust increase in EC-SOD, catalase, and eNOS with a concomitant increase in endothelial relaxation and a decrease in O2-. These observations in experimental diabetes suggest that the vascular cytoprotective mechanism of HO-1 against oxidative stress requires an increase in EC-SOD and catalase. PMID- 15821040 TI - Possible pathway(s) of testosterone egress from the active site of cytochrome P450 2B1: a steered molecular dynamics simulation. AB - To probe the possible substrate exit channel(s) in cytochrome P450 (P450) 2B1 and to clarify the role of residues previously identified by site-directed mutagenesis, a homology model was constructed based on the X-ray crystal structure of a P450 2B4-inhibitor complex. Testosterone was docked into the active site of P450 2B1 and was then pulled out through three putative channels using steered molecular dynamics simulations. The results indicated that of the three channels, the "solvent channel," lined by helices E, F, and I and the beta3 hairpin, required the largest rupture force and backbone motion, which rendered it unlikely as an exit route. The relatively small rupture forces and backbone motions for the other two channels suggested them as possible candidates for testosterone passage. The opening of channel 1, located between helices G and I and the B'-C loop, is characterized by rotation of the aromatic ring of Phe297 together with a bending of the B'-C loop. The opening of channel 2, penetrating through the B'-C loop/B' helix, is achieved by an expansion of this region and a small displacement of the backbone. Interestingly, during the egress of testosterone along channel 1, Phe297 and Phe108 appear to act as two clamps to stabilize testosterone binding and prevent it from leaving the active site. Phe115 acts as a gatekeeper for channel 2. These results are in agreement with previous site-directed mutagenesis experiments. PMID- 15821041 TI - P-glycoprotein plays a major role in the efflux of fexofenadine in the small intestine and blood-brain barrier, but only a limited role in its biliary excretion. AB - Fexofenadine is a selective, nonsedating H(1)-receptor antagonist approved for symptoms of allergic conditions, which is mainly excreted into feces via biliary excretion. The purpose of this study is to investigate its pharmacokinetics in mice and rats to determine the role of P-glycoprotein (P-gp) in its biliary excretion. In mice, biliary excretion clearance (17 ml/min/kg) accounted for almost 60% of the total body clearance (30 ml/min/kg). Comparing the pharmacokinetics after intravenous and oral administration indicated that the bioavailability of fexofenadine was at most 2% in mice. Knockout of Mdr1a/1b P-gp did not affect the biliary excretion clearance with regard to both plasma and liver concentrations, whereas the absence of P-gp caused a 6-fold increase in the plasma concentration after oral administration. In addition, the steady-state brain-to-plasma concentration ratio of fexofenadine was approximately 3-fold higher in Mdr1a/1b P-gp knockout mice than in wild-type mice. Together, these results show that P-glycoprotein plays an important role in efflux transport in the brain and small intestine but only a limited role in biliary excretion in mice. In addition, there was no difference in the biliary excretion between normal and hereditarily multidrug resistance-associated protein 2 (Mrp2) deficient mutant rats (Eisai hyperbilirubinemic rats) and between wild-type and breast cancer resistance protein (Bcrp) knockout mice. These results suggest that the biliary excretion of fexofenadine is mediated by unknown transporters distinct from P-gp, Mrp2, and Bcrp. PMID- 15821042 TI - Dextromethorphan to dextrorphan urinary metabolic ratio does not reflect dextromethorphan oral clearance. AB - Dextromethorphan urinary metabolic ratio is widely used to determine the CYP2D6 phenotype, but its utility to reflect subtle differences in catalytic activity is unclear. We evaluated the capability of dextromethorphan urinary metabolic ratio to predict dextromethorphan oral clearance as a measure of CYP2D6 activity. Data from 10 healthy extensive metabolizers of CYP2D6 were given 30 mg of dextromethorphan hydrobromide orally on two occasions. Blood and urine samples were collected for 72 h. Dextromethorphan and dextrorphan were determined in urine by high-performance liquid chromatography with fluorescence detection and in serum by liquid chromatography-mass spectrometry. The urinary metabolic ratio was very weakly correlated with dextromethorphan oral clearance (r = 0.24; p = 0.04). In contrast, the dextromethorphan oral clearance was highly correlated with the dextromethorphan to dextrorphan area under the concentration-time curve ratio (r = 0.84; p = 0.005) and the 3-h (r = 0.60; p = 0.003), 4-h (r = 0.72, p < 0.001), 6-h (r = 0.67; p < 0.001), and 8-h (r = 0.74; p < 0.001) dextromethorphan to dextrorphan serum ratios. Assuming an effect size of 30%, the number of volunteers required for crossover and cross-sectional studies using the urinary metabolic ratio as the CYP2D6 index was calculated to be 56 and 524, respectively, whereas 14 and 60 subjects are needed if oral clearance is used. Considering the required sample size and the low correlation with oral clearance, urinary metabolic ratio is not recommended as the primary outcome variable in studies requiring the detection of modest changes in CYP2D6 activity. PMID- 15821043 TI - Variable expression of MRP2 (ABCC2) in human placenta: influence of gestational age and cellular differentiation. AB - MRP2 (ABCC2) is an ATP-binding cassette (ABC)-type membrane protein involved in transport of conjugates of various drugs and endogenous compounds. MRP2 has been localized to the apical membrane of syncytiotrophoblasts and is assumed to be involved in diaplacental transfer of the above substances. It has been shown that both genetic and environmental factors can influence MRP2 expression. We therefore investigated whether gestational age, cellular differentiation, and genetic polymorphisms influence expression and localization of MRP2 in 58 human placenta samples. We detected a significant increase of transporter-mRNA with gestational age by quantitative real-time polymerase chain reaction (MRP2 mRNA/18S rRNA ratio x 1000 +/- S.D.; 0.43 +/- 0.13 in early preterms versus 1.18 +/- 0.44 in late preterms versus 2.1 +/- 0.63 in terms; p < 0.05). MRP2 protein followed the mRNA amount as shown by Western blotting (mean relative band intensity +/- S.D.; 0.56 +/- 0.1 versus 0.7 +/- 0.18 versus 0.92 +/- 0.19; early preterms versus terms p < 0.05). In cultured cytotrophoblasts, MRP2 expression increased with differentiation to syncytiotrophoblasts, with a peak on day 2 (MRP2 mRNA/18S rRNA ratio x 1000 +/- S.D.; 0.06 +/- 0.01 versus 0.88 +/- 0.27 versus 0.24 +/- 0.02 on days 0, 2, and 4). Moreover, we studied the effect of single nucleotide polymorphisms (C-24T; G1249A, and C3972T) in the MRP2 gene on placental expression. One of these polymorphisms (G1249A) resulted in a significantly reduced expression of MRP2 mRNA in preterms. In summary, the expression of MRP2 in human placenta is influenced by gestational age, cellular differentiation, and genetic factors. PMID- 15821045 TI - Contribution of CYP3A5 to hepatic and renal ifosfamide N-dechloroethylation. AB - Ifosfamide nephrotoxicity is attributed to the formation of a toxic metabolite, chloroacetaldehyde, via N-dechloroethylation, a reaction that is purportedly catalyzed by CYP3A and CYP2B6. Because allelic variants of CYP3A5 are associated with polymorphic expression of microsomal CYP3A5 in human liver and kidneys, we hypothesized that ifosfamide N-dechloroethylation depends on CYP3A5 genotype. We compared ifosfamide N-dechloroethylation activity in cDNA-expressed CYP3A4 and CYP3A5. Ifosfamide N-dechloroethylation was also assessed in liver (N = 20) and kidney (N = 21) microsomes from human donors with different CYP3A5 genotypes. Ifosfamide N-dechloroethylation was catalyzed by recombinant CYP3A5 at a rate comparable with recombinant CYP3A4. In human liver microsomes matched for CYP3A4 protein content, N-dechloroethylation was more than 2-fold higher in that from donors carrying CYP3A5*1 allele that express CYP3A5 relative to that from donors homozygous for the mutant CYP3A5*3. Correlation analysis revealed that ifosfamide N-dechloroethylation was significantly associated with CYP3A4 and CYP3A5 protein concentration but not with age, sex, or CYP2B6 protein concentration. In hepatic microsomes not expressing CYP3A5 protein, ifosfamide N-dechloroethylation was inhibited 53 to 61% and 0 to 3% by monoclonal antibodies specific for CYP3A4/5 or CYP2B6, respectively. Ifosfamide N-dechloroethylation was not detected in renal microsomes obtained from CYP3A5*3/*3 donors. In contrast, it was readily measurable in microsomes isolated from four kidneys of CYP3A5*1 carriers, which was almost completely inhibited by the CYP3A inhibitor ketoconazole. CYP2B6 protein could not be detected in this panel of human renal microsomes. In conclusion, CYP3A5*1 genotype is associated with higher rates of ifosfamide N dechloroethylation in human liver and kidneys. PMID- 15821044 TI - Lithocholic acid decreases expression of UGT2B7 in Caco-2 cells: a potential role for a negative farnesoid X receptor response element. AB - Human UDP-glucuronosyltransferase (UGT) 2B7 is the major isoform catalyzing the glucuronidation of a variety of endogenous compounds including bile acids. To determine the role of bile acids in the regulation of UGT2B7 expression, Caco-2 cells were incubated with the natural human farnesoid X receptor (hFXR) ligand, chenodeoxycholic acid, as well as the secondary bile acid, lithocholic acid, derived from chenodeoxycholic acid. Incubation of Caco-2 cells with lithocholic acid in the absence of exogenous hFXR resulted in a dose-dependent down regulation of UGT2B7 mRNA levels, with an IC(50) of 13 microM. Similar down regulation was also observed with chenodeoxycholic acid; however, much higher concentrations were required. Transient transfection of Caco-2 cells with hFXR suppressed UGT2B7 mRNA expression both in the absence and presence of ligand. UGT2B7 promoter transfection experiments and deletion/mutation analysis showed that lithocholic acid-activated hFXR decreased UGT2B7 promoter activity via a negative hFXR response element (NFRE) located between nucleotides -148 and -134. Cotransfection with hFXR and/or human retinoid X receptor further enhanced the repression. Electrophoretic mobility shift assays additionally confirmed the role of NFRE in UGT2B7 down-regulation by lithocholic acid. These findings suggest that lithocholic acid, an activator of nuclear hFXR, acts as a negative regulator of UGT2B7 expression, indicating that hFXR may play an essential role in lithocholic acid homeostasis through negative regulation of this UGT that is involved in lithocholic acid biotransformation. Therefore, it is postulated that lithocholic acid toxicity may be due to down-regulation of genes involved in its detoxification, including UGT2B7, leading to limited excretion of lithocholic acid from the body. PMID- 15821046 TI - Characterization of a novel metabolite intermediate of ziprasidone in hepatic cytosolic fractions of rat, dog, and human by ESI-MS/MS, hydrogen/deuterium exchange, and chemical derivatization. AB - Ziprasidone (Geodone), a novel atypical antipsychotic agent, is recently approved for the treatment of schizophrenia. It undergoes extensive metabolism in preclinical species and humans after oral administration, and only a very small amount of administered dose is excreted as unchanged drug. In vitro studies using human liver microsomes have shown that the oxidative metabolism of ziprasidone is mediated primarily by CYP3A4. However, coadministration of ziprasidone with ketoconazole, a CYP3A4 inhibitor, showed only a modest increase in its exposure. Therefore, in vitro metabolism of ziprasidone was investigated in hepatic cytosolic fractions to further understand its clearance mechanisms in preclinical species and humans. The major metabolite from incubation of ziprasidone in cytosolic fractions of rat, dog, and human was characterized by liquid chromatography-tandem mass spectrometry and found to be the product of reductive cleavage. Derivatization and hydrogen/deuterium exchange were used to deduce that the addition of two hydrogen atoms had occurred at the benzisothiazole moiety. Further studies to determine the enzyme involved in the formation of this metabolite are currently in progress. The identification of this novel metabolite in cytosol has clarified the clearance mechanism of ziprasidone in humans and preclinical species. PMID- 15821047 TI - Twilight for the Enlightenment? PMID- 15821048 TI - U.S. patent law. Case probes what's fair game in the search for new drugs. PMID- 15821049 TI - Infectious diseases. North Korea collaborates to fight bird flu. PMID- 15821051 TI - Livermore national lab: Settlement in bias case could unravel. PMID- 15821050 TI - Conflict of interest. Scientists, societies blast NIH ethics rules. PMID- 15821052 TI - Postdocs. Care and feeding pays off, survey finds. PMID- 15821053 TI - Postdocs. IRS takes bite out of NIH fellows' paychecks. PMID- 15821055 TI - Paleoclimatology. Cosmic dust supports a snowball Earth. PMID- 15821054 TI - Paleoanthropology. Facelift supports skull's status as oldest member of the human family. PMID- 15821056 TI - Science in Libya. From pariah to science powerhouse? PMID- 15821057 TI - Science in Libya. Evidence overruled: medics on death row. PMID- 15821058 TI - Science in Libya. Agencies plan exchange with Libya's former weaponeers. PMID- 15821059 TI - National Science Foundation. Boom in digital collections makes a muddle of management. PMID- 15821060 TI - National Science Foundation. Canadian report calls for data agency. PMID- 15821061 TI - American Physical Society meeting. Signs of a second flowing solid deepen a quantum mystery. PMID- 15821062 TI - American Physical Society meeting. In a vacuum, no one sees you splatter. PMID- 15821063 TI - American Physical Society meeting. Recipe for flies' eyes: crystallize. PMID- 15821064 TI - American Physical Society meeting. Snapshots from the meeting. PMID- 15821065 TI - Lunar and planetary science conference meeting. Ski Mars, while there's still time. PMID- 15821066 TI - Lunar and planetary science conference meeting. Rovers, dust, and a not-so-wet Mars. PMID- 15821067 TI - Lunar and planetary science conference meeting. Icy volcanism has rejuvenated Titan. PMID- 15821068 TI - Lunar and planetary science conference meeting. Snapshots from the meeting. PMID- 15821069 TI - Biodiversity in Sri Lanka and the Western Ghats. PMID- 15821070 TI - What kind of science is biology? PMID- 15821071 TI - Conduct and reporting of clinical research. PMID- 15821072 TI - Comparative studies of drug efficacy. PMID- 15821073 TI - Does the dose make the poison? PMID- 15821074 TI - Comment on "Pierolapithecus catalaunicus, a new Middle Miocene great ape from Spain". PMID- 15821076 TI - Environment. One step forward, two steps back on U.S. floodplains. PMID- 15821077 TI - Structural biology. "D" is not for diversity. PMID- 15821078 TI - Astronomy. A stellar swan-song. PMID- 15821079 TI - Psychology. Infants' insight into the mind: how deep? PMID- 15821080 TI - Chemistry. How are alkynes scrambled? PMID- 15821082 TI - Physics. Hans A. Bethe (1906-2005). PMID- 15821081 TI - Chemistry. Miniature analytical methods for medical diagnostics. PMID- 15821083 TI - Homeward sound. AB - Despite spending weeks at sea as larvae, potentially scattered over many kilometers, young coral reef fish find suitable settlement habitat and in some cases return to their natal reefs. We report that some dominant families of larval reef fish use the sounds made by fish and shrimp resident on reefs to help them locate and settle on reefs and that some fish groups use specific components of the reef sound to guide their behavior. These findings could offer potential for active management of reef fisheries. PMID- 15821084 TI - Structure of a gammadelta T cell receptor in complex with the nonclassical MHC T22. AB - Gammadelta T cell receptors (TCRs), alphabeta TCRs, and antibodies are the three lineages of somatically recombined antigen receptors. The structural basis for ligand recognition is well defined for alphabeta TCR and antibodies but is lacking for gammadelta TCRs. We present the 3.4 A structure of the murine gammadelta TCR G8 bound to its major histocompatibility complex (MHC) class Ib ligand, T22. G8 predominantly uses germline-encoded residues of its delta chain complementarity-determining region 3 (CDR3) loop to bind T22 in an orientation substantially different from that seen in alphabeta TCR/peptide-MHC. That junctionally encoded G8 residues play an ancillary role in binding suggests a fusion of innate and adaptive recognition strategies. PMID- 15821085 TI - The real-time stellar evolution of Sakurai's object. AB - After a hot white dwarf ceases its nuclear burning, its helium may briefly and explosively reignite. This causes the star to evolve back into a cool giant, whereupon it experiences renewed mass ejection before reheating. A reignition event of this kind was observed in 1996 in V4334 Sgr (Sakurai's object). Its temperature decrease was 100 times the predicted rate. To understand its unexpectedly fast evolution, we have developed a model in which convective mixing is strongly suppressed under the influence of flash burning. The model predicts equally rapid reheating of the star. Radio emission from freshly ionized matter now shows that this reheating has begun. Such events may be an important source of carbon and carbonaceous dust in the Galaxy. PMID- 15821086 TI - Formation of a carbon-carbon triple bond by coupling reactions in aqueous solution. AB - Formation of a carbon-carbon triple bond by coupling reactions usually takes place at high temperatures, in anhydrous media and anaerobic conditions. We describe the formation of a carbon-carbon triple bond at room temperature in an aqueous solution exposed to the atmosphere. Two ethylidyne ligands of a trimolybdenum cluster coupled spontaneously to form 2-butyne. This unexpected result demonstrates the plausibility of alkylidyne chain lengthening and metathesis processes under ambient, environmentally friendly conditions. PMID- 15821087 TI - A generalized approach to the modification of solid surfaces. AB - Interfacial interactions underpin phenomena ranging from adhesion to surface wetting. Here, we describe a simple, rapid, and robust approach to modifying solid surfaces, based on an ultrathin cross-linkable film of a random copolymer, which does not rely on specific surface chemistries. Specifically, thin films of benzocyclobutene-functionalized random copolymers of styrene and methyl methacrylate were spin coated or transferred, then thermally cross-linked on a wide variety of metal, metal oxide, semiconductor, and polymeric surfaces, producing a coating with a controlled thickness and well-defined surface energy. The process described can be easily implemented and adapted to other systems. PMID- 15821088 TI - Estimating duration and intensity of Neoproterozoic snowball glaciations from Ir anomalies. AB - The Neoproterozoic glaciations supposedly ended in a supergreenhouse environment, which led to rapid melting of the ice cover and precipitation of the so-called cap carbonates. If Earth was covered with ice, then extraterrestrial material would have accumulated on and within the ice and precipitated during rapid melting at the end of the glaciation. We found iridium (Ir) anomalies at the base of cap carbonates in three drill cores from the Eastern Congo craton. Our data confirm the presence of extended global Neoproterozoic glaciations and indicate that the duration of the Marinoan glacial episode was at least 3 million, and most likely 12 million, years. PMID- 15821089 TI - Vasopressin and oxytocin excite distinct neuronal populations in the central amygdala. AB - Vasopressin and oxytocin strongly modulate autonomic fear responses, through mechanisms that are still unclear. We describe how these neuropeptides excite distinct neuronal populations in the central amygdala, which provides the major output of the amygdaloid complex to the autonomic nervous system. We identified these two neuronal populations as part of an inhibitory network, through which vasopressin and oxytocin modulate the integration of excitatory information from the basolateral amygdala and cerebral cortex in opposite manners. Through this network, the expression and endogenous activation of vasopressin and oxytocin receptors may regulate the autonomic expression of fear. PMID- 15821090 TI - Antigen recognition determinants of gammadelta T cell receptors. AB - The molecular basis of gammadelta T cell receptor (TCR) recognition is poorly understood. Here, we analyze the TCR sequences of a natural gammadelta T cell population specific for the major histocompatibility complex class Ib molecule T22. We find that T22 recognition correlates strongly with a somatically recombined TCRdelta complementarity-determining region 3 (CDR3) motif derived from germ line-encoded residues. Sequence diversity around these residues modulates TCR ligand-binding affinities, whereas V gene usage correlates mainly with tissue origin. These results show how an antigen-specific gammadelta TCR repertoire can be generated at a high frequency and suggest that gammadelta T cells recognize a limited number of antigens. PMID- 15821091 TI - Do 15-month-old infants understand false beliefs? AB - For more than two decades, researchers have argued that young children do not understand mental states such as beliefs. Part of the evidence for this claim comes from preschoolers' failure at verbal tasks that require the understanding that others may hold false beliefs. Here, we used a novel nonverbal task to examine 15-month-old infants' ability to predict an actor's behavior on the basis of her true or false belief about a toy's hiding place. Results were positive, supporting the view that, from a young age, children appeal to mental states- goals, perceptions, and beliefs--to explain the behavior of others. PMID- 15821092 TI - Assortative mating in sympatric host races of the European corn borer. AB - Although a growing body of work supports the plausibility of sympatric speciation in animals, the practical difficulties of directly quantifying reproductive isolation between diverging taxa remain an obstacle to analyzing this process. We used a combination of genetic and biogeochemical markers to produce a direct field estimate of assortative mating in phytophagous insect populations. We show that individuals of the same insect species, the European corn borer Ostrinia nubilalis, that develop on different host plants can display almost absolute reproductive isolation-the proportion of assortative mating was >95%-even in the absence of temporal or spatial isolation. PMID- 15821093 TI - The floral regulator LEAFY evolves by substitutions in the DNA binding domain. AB - The plant-specific transcription factor LEAFY controls general aspects of the life cycle in a basal plant, the moss Physcomitrella patens. In contrast, LEAFY has more specialized functions in angiosperms, where it specifically induces floral fate during the reproductive phase. This raises the question of a concomitant change in the biochemical function of LEAFY during the evolution of land plants. We report that the DNA binding domain of LEAFY, although largely conserved, has diverged in activity. On the contrary, other, more rapidly evolving portions of the protein have few effects on LEAFY activity. PMID- 15821094 TI - Bivoltinism as an antecedent to eusociality in the paper wasp genus Polistes. AB - To learn the evolutionary trajectories of caste differentiation in eusocial species is a major goal of sociobiology. We present an explanatory framework for caste evolution in the eusocial wasp genus Polistes (Vespidae), which is a model system for insect eusocial evolution. We hypothesize that Polistes worker and gyne castes stem from two developmental pathways that characterized the bivoltine life cycle of a solitary ancestor. Through individual-based simulations, we show that our mechanistic framework can reproduce colony-level characteristics of Polistes and, thereby, that social castes can emerge from solitary regulatory pathways. Our explanatory framework illustrates, by specific example, a changed perspective for understanding insect social evolution. PMID- 15821095 TI - The structure of a retinal-forming carotenoid oxygenase. AB - Enzymes that produce retinal and related apocarotenoids constitute a sequence- and thus structure-related family, a member of which was analyzed by x-ray diffraction. This member is an oxygenase and contains an Fe2+-4-His arrangement at the axis of a seven-bladed beta-propeller chain fold covered by a dome formed by six large loops. The Fe2+ is accessible through a long nonpolar tunnel that holds a carotenoid derivative in one of the crystals. On binding, three consecutive double bonds of this carotenoid changed from a straight all-trans to a cranked cis-trans-cis conformation. The remaining trans bond is located at the dioxygen-ligated Fe2+ and cleaved by oxygen. PMID- 15821096 TI - Experimental and bioinformatic approaches for interrogating protein-protein interactions to determine protein function. AB - An ambitious goal of proteomics is to elucidate the structure, interactions and functions of all proteins within cells and organisms. One strategy to determine protein function is to identify the protein-protein interactions. The increasing use of high-throughput and large-scale bioinformatics-based studies has generated a massive amount of data stored in a number of different databases. A challenge for bioinformatics is to explore this disparate data and to uncover biologically relevant interactions and pathways. In parallel, there is clearly a need for the development of approaches that can predict novel protein-protein interaction networks in silico. Here, we present an overview of different experimental and bioinformatic methods to elucidate protein-protein interactions. PMID- 15821097 TI - Linking the ubiquitin-proteasome pathway to chromatin remodeling/modification by nuclear receptors. AB - Over 25 years ago, eukaryotic cells were shown to contain a highly specific system for the selective degradation of short-lived proteins, this system is known as the ubiquitin-proteasome pathway. In this pathway, proteins are targeted for degradation by covalent modification by a small highly conserved protein named ubiquitin. Ubiquitin-mediated degradation of regulatory proteins plays an important role in numerous cell processes, including cell cycle progression, signal transduction and transcriptional regulation. Recent experiments have shown that the ubiquitin-proteasome pathway is also involved in nuclear hormone receptor (NR)-mediated transcriptional regulation. The idea that the ubiquitin proteasome pathway is involved in NR-mediated transcription is strengthened by experiments showing that ubiquitin-proteasome components are recruited to NR target gene promoters. However, it is not clear how these components modulate NR mediated chromatin remodeling and gene expression. In this review, we postulate the role of the ubiquitin-proteasome pathway on NR-mediated chromatin remodeling and gene regulation based on the current knowledge from studies implicating the pathway in chromatin structure modifications that are applicable to NR function. Since evidence from this laboratory, using the glucocorticoid receptor responsive mouse mammary tumor virus (MMTV) promoter organized as chromatin, suggest that the ubiquitin-proteasome system may be involved in the elongation phase of transcription, we particularly concentrate on chromatin modifications associated with the elongation phase. PMID- 15821098 TI - Analysis of gene expression profiles in insulin-sensitive tissues from pre diabetic and diabetic Zucker diabetic fatty rats. AB - Insulin resistance occurs early in the disease process, preceding the development of type 2 diabetes. Therefore, the identification of molecules that contribute to insulin resistance and leading up to type 2 diabetes is important to elucidate the molecular pathogenesis of the disease. To this end, we characterized gene expression profiles from insulin-sensitive tissues, including adipose tissue, skeletal muscle, and liver tissue of Zucker diabetic fatty (ZDF) rats, a well characterized type 2 diabetes animal model. Gene expression profiles from ZDF rats at 6 weeks (pre-diabetes), 12 weeks (diabetes), and 20 weeks (late-stage diabetes) were compared with age- and sex-matched Zucker lean control (ZLC) rats using 5000 cDNA chips. Differentially regulated genes demonstrating > 1.3-fold change at age were identified and categorized through hierarchical clustering analysis. Our results showed that while expression of lipolytic genes was elevated in adipose tissue of diabetic ZDF rats at 12 weeks of age, expression of lipogenic genes was decreased in liver but increased in skeletal muscle of 12 week old diabetic ZDF rats. These results suggest that impairment of hepatic lipogenesis accompanied with the reduced lipogenesis of adipose tissue may contribute to development of diabetes in ZDF rats by increasing lipogenesis in skeletal muscle. Moreover, expression of antioxidant defense genes was decreased in the liver of 12-week old diabetic ZDF rats as well as in the adipose tissue of ZDF rats both at 6 and 12 weeks of age. Cytochrome P450 (CYP) genes were also significantly reduced in 12 week old diabetic liver of ZDF rats. Genes involved in glucose utilization were downregulated in skeletal muscle of diabetic ZDF rats, and the hepatic gluconeogenic gene was upregulated in diabetic ZDF rats. Genes commonly expressed in all three tissue types were also observed. These profilings might provide better fundamental understanding of insulin resistance and development of type 2 diabetes. PMID- 15821099 TI - Expression of the human melanocortin-4 receptor gene is controlled by several members of the Sp transcription factor family. AB - The melanocortin-4 receptor (MC4-R) plays a key role in the hypothalamic control of food intake, lending importance to the understanding of the mechanisms that regulate its expression. To identify factors controlling the expression of the human (h) MC4-R gene, a fragment containing 1253 bp of the 5'-flanking region of the hMC4-R gene was isolated. A series of hMC4-R luciferase constructs were developed and used to transiently transfect HEK293 and GT1-7 cell lines, both expressing endogenous MC4-R mRNA. Deletion analysis of the 1253 bp fragment showed that the basal promoter activity is mainly restricted to the 179 bp upstream of the transcription start site in both cell types. Mutation of a putative Sp1-binding site located at position -76 bp resulted in a dramatic reduction of the luciferase activity in HEK293 and GT1-7 cells by 87 and 80% respectively. Both in vitro and in vivo studies (gel shift and chromatin immunoprecipitation analyses) revealed binding of both Sp1 and Sp3 to this site in HEK293 cells. Cotransfection with an Sp1 expression vector in Drosophila cells that do not express Sp1, in conjunction with treatment of HEK293 cells with mithramycin A, a specific inhibitor of Sp1, confirmed the role of Sp1. For the first time, we have demonstrated that the constitutive activity of the hMC4-R promoter is dependent upon Sp transcription factors. PMID- 15821100 TI - The -514 C/T polymorphism in the hepatic lipase gene promoter is associated with insulin sensitivity in a healthy young population. AB - Impaired insulin action has been associated with diabetes, dyslipidemia and atherosclerotic vascular disease. The expression of insulin resistance results from the interaction of environmental and genetic factors. Human hepatic lipase (HL) is a lipolytic enzyme that plays a role in the metabolism of several lipoproteins, while insulin up-regulates the activity of HL via insulin responsive elements in the HL promoter. We have examined the influence of -514 C/T polymorphism in the hepatic lipase gene promoter on insulin sensitivity in 59 healthy young subjects (30 males and 29 females). The volunteers were subjected to three dietary periods, each lasting four weeks. During the first period all subjects consumed a saturated fat (SFA)-enriched diet with 38% as fat (20% SFA, 12% monounsaturated fatty acids (MUFA) and 6% polyunsaturated fatty acids (PUFA)). In the second and third dietary periods, a randomized crossover design was used, consisting of a low fat, high carbohydrate diet (CHO diet) (< 10% SFA, 12% MUFA and 6% PUFA) and a high-MUFA, or Mediterranean diet, with < 10% SFA, 22% MUFA and 6% PUFA. We determined the in vivo insulin resistance using the insulin suppression test with somatostatin. Steady-state plasma glucose (SSPG) concentrations (a measure of insulin sensitivity) were significantly higher in men carriers of the -514T allele after the consumption of the SFA diet than after the CHO diet and the Mediterranean diet. This effect was not observed in women. Moreover, there were no significant differences in insulin sensitivity after the three diets in men and women with the CC genotype. In summary, our results show an improvement in insulin sensitivity in men with the -514T allele of the HL promoter polymorphism, when MUFA and carbohydrates are consumed instead of SFA fat. PMID- 15821101 TI - Mechanism of protein tyrosine phosphatase 1B-mediated inhibition of leptin signalling. AB - Upon leptin binding, the leptin receptor is activated, leading to stimulation of the JAK/STAT signal transduction cascade. The transient character of the tyrosine phosphorylation of JAK2 and STAT3 suggests the involvement of protein tyrosine phosphatases (PTPs) as negative regulators of this signalling pathway. Specifically, recent evidence has suggested that PTP1B might be a key regulator of leptin signalling, based on the resistance to diet-induced obesity and increased leptin signalling observed in PTP1B-deficient mice. The present study was undertaken to investigate the mechanism by which PTP1B mediates the cessation of the leptin signal transduction. Leptin-induced activation of a STAT3 responsive reporter was dose-dependently inhibited by co-transfection with PTP1B. No inhibition was observed when a catalytically inactive mutant of PTP1B was used or when other PTPs were co-transfected. PTP1B was able to dephosphorylate activated JAK2 and STAT3 in vitro, whereas either no or a minimal effect was observed with cluster of differentiation 45 (CD45), PTPalpha and leukocyte antigen-related (LAR). By utilisation of a selective PTP1B inhibitor, the leptin induced STAT3 activation was enhanced in cells. In conclusion, these results suggested that the negative regulatory role of PTP1B on leptin signalling is mediated through a direct and selective dephosphorylation of the two signalling molecules, JAK2 and STAT3. PMID- 15821102 TI - Salivary leptin induces increased expression of growth factors in oral keratinocytes. AB - We investigated the influence of salivary leptin, purified by affinity chromatography, on the proliferation of human oral keratinocytes. Furthermore we determined the time- and dose-dependency of the incubation with salivary leptin on the production of epidermal growth factor (EGF) and keratinocyte growth factor (KGF), which are growth factors relevant to keratinocyte proliferation. The analysis was performed both intra- and extracellularly. The relationship between the three cytokines in cell proliferation was studied by successive blocking with specific antibodies. The incubation of oral keratinocytes with recombinant and native leptin led to a significantly increased, dose-dependent cell proliferation (P<0.001). A further significant increase in proliferation was observed after incubating the cells with sterile filtered saliva (P<0.001). The increase in proliferation could not be observed by simultaneous incubation with salivary leptin and specific antibodies against either leptin or KGF (P<0.001). We found a significant dose-dependency between leptin incubation and production of KGF and EGF at the RNA and protein level. Both cytokines were expressed intracellularly and released into the culture medium, where they could be quantified by ELISA. Furthermore, there was a dose- and time-dependent increase in the phosphorylation of STAT-1 and STAT-3, indicating that Ob-R(b) (the long form of the leptin receptor) expressed by the keratinocytes is functional. It is conceivable that the leptin-induced proliferation in keratinocytes is mediated by this signalling pathway. This is the first study to show a physiological role of salivary leptin as a growth factor for keratinocyte proliferation in the oral cavity. We could demonstrate its influence on the production of other growth factors essential for this proliferative effect. Based on the findings of our study we assume an important role for salivary leptin in wound healing within the vulnerable oral cavity. PMID- 15821103 TI - The acquisition of an insulin-secreting phenotype by HGF-treated rat pancreatic ductal cells (ARIP) is associated with the development of susceptibility to cytokine-induced apoptosis. AB - The elucidation of mechanisms regulating the regeneration and survival of pancreatic beta cells has fundamental implications in the cell therapy of type 1 diabetes. The present study had the following three aims: 1. to investigate whether pancreatic ductal epithelial cells can be induced to differentiate into insulin-producing cells by exposing them to hepatocyte growth factor (HGF); 2. to characterize some of the molecular events leading to their differentiation toward a beta-cell-like phenotype; 3. to evaluate the susceptibility of newly differentiated insulin-secreting cells to cytokine-induced apoptosis, a mechanism of beta-cell destruction occurring in type 1 diabetes. We demonstrated that HGF treated rat pancreatic ductal cell line (ARIP) cells acquired the capability to transcribe the insulin gene and translate its counterpart protein. HGF-treated cells also exhibited a glucose-dependent capability to secrete insulin into the cultured medium. Expression analysis of some of the genes regulating pancreatic beta-cell differentiation revealed a time-dependent transcription of neurogenin-3 and Neuro-D in response to HGF. Finally, we determined the susceptibility to proinflammatory cytokine (PTh1)-induced apoptosis by incubating HGF-treated and untreated ARIP cells with a cocktail of interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). Such treatment induced apoptotic death, as determined by the TUNEL technique, in about 40% of HGF-treated, insulin-secreting ARIP cells, while untreated ARIP cells were resistant to PTh1-induced apoptosis. In conclusion, we showed that HGF promotes the differentiation of ARIP cells into pancreatic beta-cell-like cells, and that the differentiation toward an insulin-secreting phenotype is associated with the appearance of susceptibility to cytokine-induced apoptosis. PMID- 15821104 TI - Pancreatic beta-cells expressing GLP-1 are resistant to the toxic effects of immunosuppressive drugs. AB - Glucose intolerance is often observed after pancreatic islet cell transplantation. The administration of immunosuppressive agents (ISD), necessary to avoid tissue rejection, is in part responsible for hyperglycemia. To investigate whether mouse insulinoma (MIN6) cells transfected with the glucagon like peptide-1 (GLP-1) fragment of the proglucagon gene (RIP/GLP-1 MIN6 cells) are resistant to the toxicity derived from the administration of ISD. RIP/GLP-1 MIN6 cells, as well as parental MIN6 cells, were exposed to a cocktail of ISD. The secretion of insulin and the expression of apoptosis-related proteins were investigated by RIA and western blot analysis. Cell apoptosis was quantified by FACS analysis. Finally, to study whether the antiapoptotic action of GLP-1 was a function of its effect on insulin secretion, or rather it was a direct effect of GLP-1, cells were cultured with or without diazoxide or exendin-9. GLP-1 improved the functional activity and the viability of cells exposed to ISD. The insulin secretion of RIP/GLP-1 MIN6 cells after exposure to ISD was preserved. The expression of GLP-1 by beta-cells reduced the number of apoptotic cells and increased the expression of the antiapoptotic protein Bcl-2. GLP-1 also decreased the abundance of the proapoptotic markers PARP-p85 and Smac/Diablo. Treatment of cells with the diazoxide did not abolish the protective advantage that cells transfected with GLP-1 had; conversely the exposure of cells to exendin-9 was associated with a restored susceptibility to apoptosis. This report demonstrates that GLP-1 is capable of preserving beta-cell function and protecting cells from apoptotic cell death. PMID- 15821105 TI - Tetracycline-regulated secretion of human (pro)insulin following plasmid-mediated transfection of human muscle. AB - Long-term secretion of insulin by host muscle following transduction with an insulin gene construct offers the potential of gene therapy for diabetes without immunosuppression. Clinical implementation will be dependent on proof of principle in human tissue and a system for safely regulating basal insulin levels. Liposomal co-transfection with a tetracycline-responsive wild type human preproinsulin (pTRE-hppI1) or mutant construct (pTRE-hppI4), in which PC2 and PC3 cleavage sites were altered to form tetrabasic consensus sites for furin, together with pTet-off (coding for a transactivating protein) was evaluated in the C2C12 mouse myoblast cell line and human myoblasts following establishment in primary culture. In the absence of tetracycline, (pro)insulin secretion in C2C12 and human myoblasts transfected with tetracycline-responsive hppI1 and hppI4 constructs was comparable to that following transfection with equivalent constructs under the control of a constitutively active cytomegaloviral promoter. Percentage processing to mature insulin was <5% in C2C12 and human myoblasts transfected with pTet-off/pTRE-hppI1 but >90% in C2C12 cells and 45-60% in human myoblasts on transfection with pTet-off/pTRE-hppI4. Incremental dose-responsive suppression of proinsulin secretion was demonstrated in C2C12 and human myoblasts expressing pTet-off/pTRE-hppI1 following incubation with tetracycline (0-100 microg/ml) for up to 72 h. Reversibility was confirmed following tetracycline withdrawal. Dose-responsive tetracycline-inducible repression of mature insulin secretion was confirmed in C2C12 cells following transfection with pTet-off/pTRE hppI4. Regulation of human proinsulin biosynthesis and secretion has been attained in vivo following plasmid-mediated gene transfer to rat skeletal muscle and oral tetracycline administration. In conclusion, processing to mature insulin has been confirmed following plasmid-mediated gene transfer to human muscle in addition to in vitro- and in vivo-regulated human proinsulin secretion employing the safe and well-tolerated antibiotic, tetracycline. PMID- 15821106 TI - Effect of inactivating mutations on phosphorylation and internalization of the human VPAC2 receptor. AB - The VPAC(2) receptor, as all members of the G-protein-coupled receptor (GPCR)-B family, has two highly conserved motifs in the third intracellular (IC(3)) loop: a lysine and a leucine located at the amino-terminus and two basic residues separated by a leucine and an alanine at the carboxyl-terminus. This study evaluates the involvement of those conserved amino acid sequences in VPAC(2) signal transduction and regulation. The residues were mutated into alanine and mutants were expressed in Chinese hamster ovary (CHO) cells stably transfected with Galpha16 and aequorin. Mutation of L310 reduced efficacy of vasoactive intestinal polypeptide (VIP) to stimulate adenylate cyclase activity through Galphas coupling by 75%, without affecting VIP capability to stimulate an increase in [Ca(2+)](i) through Galpha16 coupling. Mutation of R325 and, to a lesser extend, K328 reduced VIP efficacy to stimulate [Ca(2+)](i) increase and VIP potency to stimulate adenylate cyclase. The combination of mutations of both amino- and carboxyl-terminus located conserved motifs of the IC(3) loop generates an inactive receptor with respect to [Ca(2+)](i) increase and adenylate cyclase activation, but also with respect to receptor phosphorylation and internalization that were indeed directly correlated with the potency of inactivation of the receptors. The amino-terminus of the VPAC(2) receptor IC(3) loop is thus involved in adenylate cyclase activation and the carboxyl-terminus of the IC(3) loop participates in both Galphas and Galpha16 coupling. The mutations studied also reduced both receptor phosphorylation and internalization in a manner that appeared directly linked to the alteration of Galphas and Galpha16 coupling. PMID- 15821107 TI - Paracrine regulation of growth hormone gene expression by gonadotrophin release in grass carp pituitary cells: functional implications, molecular mechanisms and signal transduction. AB - Growth hormone (GH) is known to stimulate luteinizing hormone (LH) release via paracrine interactions between somatotrophs and gonadotrophs. However, it is unclear if LH can exert a reciprocal effect to modulate somatotroph functions. Here we examined the paracrine effects of LH on GH gene expression using grass carp pituitary cells as a cell model. LH receptors were identified in grass carp somatotrophs and their activation by human chorionic gonadotropin (hCG) increased 'steady-state' GH mRNA levels. Removal of endogenous LH by immunoneutralization using LH antiserum inhibited GH release and GH mRNA expression. GH secretagogues, including gonadotrophin releasing hormone (GnRH), pituitary adenylate cyclase activating polypeptide (PACAP) and apomorphine, were effective in elevating GH mRNA levels but these stimulatory actions were blocked by LH antiserum. In pituitary cells pretreated with actinomycin D, the half-life of GH mRNA was not affected by hCG but was enhanced by LH immunoneutralization. Treatment with LH antiserum also suppressed basal levels of mature GH mRNA and primary transcripts. hCG increased cAMP synthesis in carp pituitary cells and hCG-induced GH mRNA expression was mimicked by forskolin but suppressed by inhibiting adenylate cyclase and protein kinase A. Similarly, the stimulatory actions of hCG and forskolin on GH mRNA expression were blocked by inhibiting Janus kinase 2 (JAK2) and MAP kinase (MAPK), including P42/44(MAPK) and P38 (MAPK). These results suggest that LH is essential for the maintenance of GH release, GH gene expression, and somatotroph responsiveness to GH-releasing factors. The paracrine actions of LH on GH mRNA expression are mediated by a concurrent increase in GH gene transcription and GH mRNA turnover, probably through JAK2/MAPK coupled to the cAMP-dependent pathway. PMID- 15821108 TI - Gene regulation by NMDA receptor activation in the SDN-POA neurons of male rats during sexual development. AB - The present study was designed to identify possible signaling pathways, which may play a role in prevention of neuronal apoptosis in the sexually dimorphic nucleus of the preoptic area (SDN-POA) after physiological activation of the N-methyl-D aspartate (NMDA) receptor. Gene response to the blockage of the NMDA receptor by an antagonist (dizocilpine hydrogen maleate; MK-801) was screened after suppression subtractive hybridization (SSH). The results showed that differential screening after SSH detected the presence of some neurotrophic genes (RNA binding motif protein 3 (RBM3), alpha-tubulin) as well as apoptosis-related genes (Bcl-2, cytochrome oxidase subunit II, cytochrome oxidase subunit III) in the SDN-POA of male rats, which were down-regulated by blocking the NMDA receptor. The RT-PCR products of the aforementioned genes in MK-801-treated males were significantly less than that in untreated males. In particular, the expression of Bcl-2 mRNA, including Bcl-2 protein, in male rats were significantly suppressed by MK-801 treatment. Moreover, the binding activity of nuclear factor kappaB (NFkappaB) was significantly higher in male rats than in females, but significantly diminished by blocking the NMDA receptor with MK-801 in male rats. No significant difference in cAMP response element-binding protein (CREB) binding activity was observed among untreated male, MK-801-treated male, untreated female and MK-801-treated female groups. These results suggest that genes regulated by NMDA receptor activation might participate in neuronal growth and/or anti-apoptosis, and support an important signaling pathway of NFkappaB activation and its target gene, Bcl-2, in preventing neuronal apoptosis in the SDN-POA of male rats during sexual development. PMID- 15821109 TI - The post-endocytotic fate of the gonadotropin receptors is an important determinant of the desensitization of gonadotropin responses. AB - Internalization of the ligand/receptor complexes is a consequence of the activation of the gonadotropin receptors. Since the recycling or degradation of the internalized receptors results in the maintenance or loss of cell surface receptors respectively and this contributes to the loss of responsiveness, we hypothesized that the fate of the internalized receptors could be an important component of desensitization. We examined this hypothesis using the wild-type and mutants of the human LH (hLHR) receptors and follitropin receptors expressed in MA-10 and KK-1 cells respectively. The receptor mutants were chosen because they are routed mostly to a lysosomal degradation pathway whereas the wild-type receptors are recycled back to the surface. We have shown that agonist stimulation of cells expressing the mutant receptors results in a more pronounced loss of cell surface receptors and agonist responses than stimulation of cells expressing the wild-type receptors. We concluded that receptor recycling promotes the maintenance of cell surface receptors and preserves hormonal responsiveness. This property of the hLHR is likely to be physiologically important because there at least two hLHR-expressing tissues in pregnant women, the maternal corpus luteum and the fetal Leydig cells, where a loss of hormonal responsiveness induced by the elevated levels of human chorionic gonadotropin that occur during pregnancy is not desirable. PMID- 15821110 TI - Atrial natriuretic peptide inhibits the actions of FSH and forskolin in meiotic maturation of pig oocytes via different signalling pathways. AB - Atrial natriuretic peptide (ANP) as well as its receptors is found in mammalian ovary and follicular cells and its function in oocyte meiotic maturation has also been reported in Xenopus, hamster and rat. But the results are controversial and the physiological mechanism of ANP on oocyte maturation is not clear, especially the relationship between gonadotrophin and ANP as well as the signal transduction, and these need further study. The present study conducted experiments to examine these questions by using drug treatment and Western blot analysis and focused on pig oocyte meiotic maturation and cumulus expansion in vitro. The results revealed that ANP could inhibited FSH-induced pig oocyte maturation and cumulus expansion and prevent the full phosphorylation of mitogen activated protein kinase in both oocytes and cumulus cells, and that these inhibitory effects could be mimicked by 8-Br-cyclic guanosine 5'-monophosphate (8 Br-cGMP), but blocked by a protein kinase G (PKG) inhibitor KT5823. Zaprinast, a cGMP-specific phosphodiesterase inhibitor, could enhance the inhibitory effect of ANP on oocyte maturation. A specific analogue of ANP, C-ANP-(4-23), which binds to the natriuretic peptide receptor-C (NPRC), had no effect in either FSH-induced or spontaneous oocyte maturation. Treatment with forskolin, a stimulator of adenylate cyclase, had a biphasic effect; 44 h treatment induced cumulus expansion but inhibited oocyte maturation while 2 h treatment induced maturation of cumulus-enclosed oocytes (CEOs). Both ANP and C-ANP-(4-23) could inhibit the effect of forskolin on CEO maturation, and these inhibitory effects of ANP/C-ANP (4-23) could be blocked by preincubation with pertussis toxin (PT), consistent with mediation by a Gi protein(s) in the cumulus cells. All these results suggest that ANP is a multifunctional regulator of FSH and forskolin on pig CEO maturation by two signalling mechanisms: one is via a cGMP/PKG pathway, the other is via NPRC receptors in cumulus cells and the activation of the PT-sensitive Gi protein(s). PMID- 15821111 TI - Estrogen-related receptor-gamma and peroxisome proliferator-activated receptor gamma coactivator-1alpha regulate estrogen-related receptor-alpha gene expression via a conserved multi-hormone response element. AB - The expression of estrogen-related receptor-alpha (ERRalpha) is stimulated by estrogen in selective tissues. Recently, a correlation between ERRalpha expression and the induction of peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) in the liver of fasting animals and in cold stressed brown-fat tissues and skeletal muscle was shown. To explore the molecular mechanisms of ERRalpha regulation by diverse signals, the promoter of the human ERRalpha gene was cloned and characterized. Mutation and deletion analyses revealed that a 53 bp region containing repeated core element AGGTCA motifs of the ERRalpha gene serves as a multi-hormone response element (MHRE) for several nuclear receptors in transient co-transfection studies of human endometrial carcinoma (HEC-1B) cells. Among the nuclear receptors tested, ERRgamma bound to and robustly stimulated the transcription of reporters containing at least two AGGTCA motifs. Ectopic expression of PGC-1alpha in HEC-1B cells strongly activated the reporter containing the MHRE, presumably via the endogenous nuclear receptor binding to the element. Reducing the endogenous level of ERRgamma by small interfering RNA, and increasing the ERRgamma level by ectopic expression, substantially decreased and increased respectively the transactivation capability of PGC-1alpha. The activation function 2 domain of the ERRgamma and the L2 and L3 motifs of PGC-1alpha were essential to transactivate the MHRE. Additionally, PGC-1alpha increases the amount of endogenous ERRgamma bound to the MHRE region as determined by a chromatin immunoprecipitation assay. The present study demonstrates that the MHRE of the ERRalpha gene is a target for ERRgamma transactivation, which is enhanced by PGC-1alpha. PMID- 15821112 TI - Gonadal defects and hormonal alterations in transgenic mice expressing a single chain human chorionic gonadotropin-lutropin receptor complex. AB - To study the effects of premature and chronic ligand-mediated luteinizing hormone receptor (LHR) activation on reproductive development, we have generated transgenic mice expressing a genetically engineered, constitutively active yoked hormone-receptor complex (YHR), in which a fusion protein of human chorionic gonadotropin (hCG) is covalently linked to the N-terminus of rat LHR. YHR expressing mice (YHR(+)) were analyzed at pre- and post-pubertal ages. Relative to wild type (WT) controls, male mice exhibited prepubertal increases in testosterone levels and seminal vesicle weights, and decreases in serum FSH, serum LH, testes weight, and the size of the seminiferous tubules. In adult male YHR(+) mice, testosterone and LH levels are not significantly different from WT controls. However, FSH levels and testes weights remain decreased. Female YHR(+) mice undergo precocious puberty with early vaginal opening, accelerated uterine development, enhanced follicular development, including the presence of corpora lutea, and an increase in serum progesterone. At 12 weeks of age, the ovary exhibits a relative increase in the amount of interstitial tissue, comprised of cells that are hypertrophic and luteinized, as well as follicles that are degenerating. Additionally, hemorrhagic cysts develop in approximately 25% of the transgenic mice. These degenerative changes are consistent with an aging ovary suggesting that CG-induced LHR activation in female mice leads to precocious sexual development and ovarian lesions. Taken together, these data indicate that the single chain YHR is functional in vivo and demonstrate that YHR(+) mice provide a novel system to further understand the reproductive consequences of aberrant LHR activation. PMID- 15821113 TI - betaA- and betaC-activin, follistatin, activin receptor mRNA and betaC-activin peptide expression during rat liver regeneration. AB - The mRNA expression of two activin growth factor subunits (betaA- and betaC activin), activin receptor subunits (ActRIIA, ActRIIB) and the activin-binding protein follistatin, and peptide expression of betaA-activin and betaC-activin subunits, were examined in regenerating rat liver after partial hepatectomy (PHx). Liver samples were collected from adult, male Sprague-Dawley rats, 12-240 h (n=3-5 rats per time point) after PHx or from sham-operated controls at the same time points. Hepatocyte mitosis and apoptosis were assessed histologically and by in situ cell death detection. RT and PCR were used to assess relative gene expression. betaA- and betaC-activin peptide immunoreactivity was assessed in liver and serum samples by western blotting, whereas cellular expression was investigated by immunohistochemistry, using specific monoclonal antibodies. betaA and betaC-activin mRNA dropped to < 50% of sham control values 12 h after PHx and remained at this level until 168 h post-PHx, when betaA-activin expression increased to three times sham control values and betaC-activin mRNA returned to pre-PHx levels. A peak in follistatin expression was observed 24-48 h post-PHx, coincident with an increase in hepatocyte mitosis. No changes were observed in ActRIIA mRNA, whereas ActRIIB expression paralleled that of betaA-activin mRNA. betaC-activin immunoreactive homo- and heterodimers were observed in regenerating liver and serum. Mitotic hepatocytes frequently contained betaC-activin immunoreactivity, whereas apoptotic hepatocytes were often immunoreactive for betaA-activin. We conclude that betaA- and betaC-activin subunit proteins are autocrine growth regulators in regenerating liver and when expressed independently lead to hepatocyte apoptosis or mitosis in a subset of hepatocytes. PMID- 15821114 TI - Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells. AB - Information on the regulation of steroid hormone receptors and their distinct functions within the human endometrial epithelium is largely unavailable. We have immortalized human primary endometrial epithelial cells (EECs) isolated from a normal proliferative phase endometrium by stably transfecting the catalytic subunit (hTERT) of the human telomerase complex and cultured these hTERT-EECs now for over 350 population doublings. Active hTERT was detected in hTERT-EECs employing the telomerase repeat amplification assay protocol. hTERT-EECs revealed a polarized, non-invasive epithelial phenotype with apical microvilli and production of a basal lamina when grown on a three-dimensional collagen fibroblast lattice. Employing atomic force microscopy, living hTERT-EECs were shown to produce extracellular matrix (ECM) components and ECM secretion was modified by estrogen and progesterone (P4). hTERT-EECs expressed inducible and functional endogenous estrogen receptor-alpha (ER-alpha) as demonstrated by estrogen response element reporter assays and induction of P4 receptor (PR). P4 treatment down-regulated PR expression, induced MUC-1 gene activity and resulted in increased ER-beta transcriptional activity. Gene activities of cytokines and their receptors interleukin (IL)-6, leukemia inhibitory factor (LIF), IL-11 and IL-6 receptor (IL6-R), LIF receptor and gp130 relevant to implantation revealed a 17 beta-estradiol (E2)-mediated up-regulation of IL-6 and an E2- and P4-mediated up-regulation of IL6-R in hTERT-EECs. Thus, hTERT-EECs may be regarded as a novel in vitro model to investigate the role of human EECs in steroid hormone-dependent normal physiology and pathologies, including implantation failure, endometriosis and endometrial cancer. PMID- 15821115 TI - Anti-proliferative effect of estrogen in breast cancer cells that re-express ERalpha is mediated by aberrant regulation of cell cycle genes. AB - Estrogen receptor (ER)-negative breast carcinomas do not respond to hormone therapy, making their effective treatment very difficult. The re-expression of ERalpha in ER-negative MDA-MB-231 breast cancer cells has been used as a model system, in which hormone-dependent responses can be restored. Paradoxically, in contrast to the mitogenic activity of 17beta-estradiol (E2) in ER-positive breast cancer cells, E2 suppresses proliferation in ER-negative breast cancer cells in which ERalpha has been re-expressed. We have used global gene expression profiling to investigate the mechanism by which E2 suppresses proliferation in MDA-MB-231 cells that express ERalpha through adenoviral infection. We show that a number of genes known to promote cell proliferation and survival are repressed by E2 in these cells. These include genes encoding the anti-apoptosis factor SURVIVIN, positive cell cycle regulators (CDC2, CYCLIN B1, CYCLIN B2, CYCLIN G1, CHK1, BUB3, STK6, SKB1, CSE1 L) and chromosome replication proteins (MCM2, MCM3, FEN1, RRM2, TOP2A, RFC1). In parallel, E2-induced the expression of the negative cell cycle regulators KIP2 and QUIESCIN Q6, and the tumour-suppressor genes E CADHERIN and NBL1. Strikingly, the expression of several of these genes is regulated in the opposite direction by E2 compared with their regulation in ER positive MCF-7 cells. Together, these data suggest a mechanism for the E2 dependent suppression of proliferation in ER-negative breast cancer cells into which ERalpha has been reintroduced. PMID- 15821116 TI - Inducible upregulation of oestrogen receptor-beta1 affects oestrogen and tamoxifen responsiveness in MCF7 human breast cancer cells. AB - To investigate the effect of altered oestrogen receptor (ER)alpha and ERbeta expression on oestrogen and anti-oestrogen action in breast cancer, we have stably expressed an inducible ERbeta1 in MCF7 breast cancer cells. Stably expressing clones were isolated and over-expression of ERbeta1 correlated with increased levels of specific radiolabelled oestradiol (E2) binding. Increased ERbeta1 did not affect endogenous levels of ERalpha but increased progesterone receptor (PR) levels. Over-expression of ERbeta1 reduced growth responses to E2 in contrast to little if any effect of over-expression of ERalpha. In oestrogen replete conditions, over-expression of ERbeta1 but not ERalpha reduced proliferation. Over-expression of ERbeta1 did not result in anti-oestrogen resistance but was associated with increased sensitivity to 4-hydroxytamoxifen. Our results suggested that over-expression of ERbeta1 in the presence of an endogenously expressed ERalpha was associated with tamoxifen sensitivity but may negatively modulate ERalpha-mediated growth. However, not all ERalpha activities were inhibited since endogenous PR expression was increased by both ERalpha and ERbeta1 over-expression. These data paralleled those seen in some in vivo studies showing a relationship between PR and ERbeta expression as well as ERbeta expression and tamoxifen sensitivity of ER-positive breast cancer patients. These models are relevant and will be useful for dissecting the role of ERbeta1 expression in ER-positive breast cancer. PMID- 15821117 TI - A conserved retinoid X receptor (RXR) from the mollusk Biomphalaria glabrata transactivates transcription in the presence of retinoids. AB - Retinoid X receptors (RXR) are members of the nuclear receptor superfamily of ligand-activated transcription factors that have been characterized in a wide variety of metazoan phyla. They act as heterodimer partners of other nuclear receptors, and in vertebrates also activate transcription as homodimers in the presence of a ligand, 9-cis retinoic acid. In order to test the hypothesis that retinoic acid signaling pathways involving RXRs are present in the Lophotrochozoa, we have sought to isolate conserved members of this family from the platyhelminth parasite Schistosoma mansoni and its intermediate host, the mollusk Biomphalaria glabrata. Here we report that an RXR ortholog from B. glabrata (BgRXR) is better conserved, compared with mouse RXRalpha, both in the DNA-binding domain (89% identity) and in the ligand-binding domain (LBD) (81% identity), than are arthropod homologs. In EMSA, BgRXR binds to the direct repeat response element DR1 as a homodimer or as a heterodimer with mammalian RARalpha, LXR, FXR or PPARalpha. When transfected alone into mammalian cell lines, BgRXR transactivated transcription of a reporter gene from the Apo-A1 promoter in the presence of 9-cis retinoic acid or DHA. Constructs with the Gal4 DNA binding domain fused to the hinge and LBDs of BgRXR were used to show that ligand dependent activation of transcription by BgRXR required its intact AF-2 activation domain, and that the LBD can form homodimers. Finally, the binding of 9-cis retinoic acid preferentially protected the LBD of BgRXR from degradation by trypsin in a proteolysis protection assay. Our results show that BgRXR binds and is activated by retinoids and suggest that retinoid signaling pathways are conserved in the Lophotrochozoa. The nucleotide sequence reported in this paper has been submitted to the GenBank/EBI Data Bank with accession no. AY048663. PMID- 15821118 TI - Glucocorticoids suppress macrophage migration inhibitory factor (MIF) expression in a cell-type-specific manner. AB - MIF is a potent proinflammatory cytokine involved in inflammatory arthritis. Glucocorticoids (GC) have been reported to induce secretion of MIF in rodent cells, and as MIF counteracts the anti-inflammatory effects of GC, this has implications for human inflammatory disease. Transient transfection studies showed that the MIF promoter was repressed by dexamethasone (Dex) (10 nM) in CEM C7A cells, with up to 50% suppression by 100 nM. However, there was no regulation of the promoter by GC in A549 cells. We also found that subnanomolar concentrations of Dex suppressed MIF secretion, measured by ELISA, by 80% in both human T lymphoblasts (CEM C7A) and human lung epithelial cells (A549). Endogenous MIF mRNA was also repressed by GC in CEM C7A cells, measured both by Northern blot and quantitative RT-PCR assays, but there was no such regulation in A549 cells. This suggests that GC affects translation rather than transcription of MIF in A549 cells. These results contradict earlier results with the rat cell line RAW 264.7. Therefore, we analysed MIF secretion from RAW 264.7 cells but found no GC effect on secretion. Understanding how GC regulates MIF in a cell-type dependent manner may give insights into GC-refractory human inflammatory diseases. PMID- 15821119 TI - Does an oral analgesic protocol improve pain control for patients with cancer? An intergroup study coordinated by the Eastern Cooperative Oncology Group. AB - BACKGROUND: Cancer pain is highly prevalent and commonly undertreated. This study was designed to determine whether dissemination of a clinical protocol for pain management would improve outcomes in community oncology practices. PATIENTS AND METHODS: A pain management protocol was developed based on accepted guidelines. After baseline assessment, oncology practices were randomly assigned to 'analgesic protocol' (AP) sites, where oncologists implemented the guidelines in a group of lung or prostate cancer patients, or to 'physician discretion' (PD) sites, where customary treatment was continued. Patients treated on protocol and a comparison group of patients with pain due to breast cancer or myeloma were monitored for change in pain using the Brief Pain Inventory, and for change in other symptoms or mood. RESULTS: The protocol terminated early because of poor accrual. We compared groups using proportions of patients who had no or mild pain at follow-up. Although measures of protocol adherence did not suggest the occurrence of major practice change, the proportion of lung or prostate cancer patients with no or mild pain increased significantly from baseline for those treated at AP sites compared with those treated at PD sites. There was no significant difference between the breast and myeloma patients treated at AP sites versus those treated at PD sites. CONCLUSION: A protocol for cancer pain management can improve pain control. Diffusion of these benefits to other patients was not confirmed. Given the small sample size, these findings require confirmation in a larger trial. PMID- 15821120 TI - First-line gemcitabine versus epirubicin in postmenopausal women aged 60 or older with metastatic breast cancer: a multicenter, randomized, phase III study. AB - BACKGROUND: This randomized, phase III study compared the efficacy and safety of first-line gemcitabine versus epirubicin in the treatment of postmenopausal women with metastatic breast cancer (MBC). PATIENTS AND METHODS: Patients aged > or = 60 years (median 68 years) with clinically measurable MBC received either gemcitabine 1200 mg/m(2) or epirubicin 35 mg/m(2) on days 1, 8, and 15 of a 28 day cycle. RESULTS: Of 410 patients entered, 397 (198 gemcitabine and 199 epirubicin) were randomized and qualified for the time to progressive disease (TTP) and survival analyses. Total cycles administered in 185 gemcitabine and 192 epirubicin patients, respectively, were 699 (mean 3.5, range 0-12) and 917 (mean 4.6, range 0-10). Epirubicin demonstrated statistically significant superiority in TTP (6.1 and 3.4 months, P=0.0001), overall survival (19.1 and 11.8 months, P=0.0004), and independently assessed response rate (40.3% and 16.4% in 186 and 183 evaluable patients, P <0.001). For gemcitabine (n=190) and epirubicin (n=192), respectively, common WHO grade 3/4 toxicities were neutropenia (25.3% and 17.9%) and leukopenia (14.3% and 19.3%). Of the 28 on-study deaths (17 gemcitabine, 11 epirubicin), three were considered possibly or probably related to treatment (gemcitabine). CONCLUSIONS: Postmenopausal women > or =60 years of age with MBC tolerate chemotherapy well. In this study, epirubicin was superior to gemcitabine in the treatment of MBC in women age > or =60, confirming that anthracyclines remain important drugs for first-line treatment of MBC. PMID- 15821121 TI - Tumour budding detected by laminin-5 {gamma}2-chain immunohistochemistry is of prognostic value in epidermoid anal cancer. AB - BACKGROUND: Markers for guidance with regard to individual prognosis and treatment planning are sought in epidermoid anal cancer. This study assessed the prognostic and predictive value of tumour budding. PATIENTS AND METHODS: From a population-based consecutive series of patients who were prospectively recorded, it was possible to investigate 209 (76%) of the pretreatment biopsies. Immunohistochemistry with a monoclonal antibody for the gamma2 chain of laminin-5 was used to detect tumour budding (defined as dissociated single cancer cells or clusters of up to five cells). RESULTS: Tumour budding was detected in 104 (50%) of the 209 samples. No significant correlation was found between tumour budding and clinicopathological characteristics. Patients with tumour budding had a statistically significantly better 5-year overall survival rate compared with patients lacking tumour budding (74% versus 64%, P <0.05). Albeit not statistically significant, other outcome variables such as tumour-specific survival, recurrence after initial complete response and rate of distant metastases, were all in favour of patients with tumour budding. Multivariate analysis reveals tumour budding as an independent positive prognostic factor. CONCLUSIONS: Tumour budding detected by laminin-5 immunohistochemistry may be of prognostic value in the treatment of epidermoid anal cancer. However, further studies are needed to clarify the possible clinical implications. PMID- 15821122 TI - Carcinoma in situ testis, the progenitor of testicular germ cell tumours: a clinical review. AB - Testicular germ cell tumours (TGCT), including seminomas, embryonal carcinomas, teratomas and yolk sac tumours, have a common precursor, the carcinoma in situ (CIS) cell. Recent gene expression studies displaying close similarity of CIS cells to embryonic stem cells support the longstanding theory that CIS most likely originates in utero from fetal gonocytes. The clinical association between the testicular dysgenesis syndrome components (TGCT, cryptorchidism, genital malformations, some forms of decreased spermatogenesis) also implies a prenatal origin. Despite high cure rates of TGCT, efforts should be made to obtain diagnosis at the CIS stage, as intervention is possible before an invasive tumour develops, thus reducing the necessity for intensive therapy. CIS may be suspected in patients with an assumed extragonadal GCT or cryptorchidism, and in intersex patients and selected cases with infertility (presenting with atrophic testes and ultrasonic microlithiasis). Surgical testicular biopsy seems the only reliable diagnostic method. The management of choice of unilateral CIS is orchidectomy, or localised irradiation in bilateral cases. At least 5% of TGCT patients present with contralateral CIS; therefore, contralateral biopsy is recommended at the time of orchidectomy. Further research is warranted to identify causal factors explaining the increasing incidence of TGCT and to obtain a method of non invasive CIS detection. PMID- 15821123 TI - Candida biofilms: an update. PMID- 15821124 TI - Surface hydrophobicity changes of two Candida albicans serotype B mnn4delta mutants. AB - Cell surface hydrophobicity (CSH) of Candida species enhances virulence by promoting adhesion to host tissues. Biochemical analysis of yeast cell walls has demonstrated that the most significant differences between hydrophobic and hydrophilic yeasts are found in the acid-labile fraction of Candida albicans phosphomannoprotein, suggesting that this fraction is important in the regulation of the CSH phenotype. The acid-labile fraction of C. albicans is unique among fungi, in that it is composed of an extended polymer of beta-1,2-mannose linked to the acid-stable region of the N-glycan by a phosphodiester bond. C. albicans serotype A and B strains both contain a beta-1,2-mannose acid-labile moiety, but only serotype A strains contain additional beta-1,2-mannose in the acid-stable region. A knockout of the C. albicans homolog of the Saccharomyces cerevisiae MNN4 gene was generated in two serotype B C. albicans patient isolates by using homologous gene replacement techniques, with the anticipation that they would be deficient in the acid-labile fraction and, therefore, demonstrate perturbed CSH. The resulting mnn4delta-deficient derivative has no detectable phosphate-linked beta-1,2-mannose in its cell wall, and hydrophobicity is increased significantly under conditions that promote the hydrophilic phenotype. The mnn4delta mutant also demonstrates an unanticipated perturbation in the acid-stable mannan fraction. The present study reports the first genetic knockout constructed in a serotype B C. albicans strain and represents an important step for dissecting the regulation of CSH. PMID- 15821125 TI - Combinatorial repression of the hypoxic genes of Saccharomyces cerevisiae by DNA binding proteins Rox1 and Mot3. AB - The hypoxic genes of Saccharomyces cerevisiae are transcriptionally repressed during aerobic growth through recruitment of the Ssn6/Tup1 general repression complex by the DNA binding protein Rox1. A second DNA binding protein Mot3 enhances repression of some hypoxic genes. Previous studies characterized the role of Mot3 at the hypoxic ANB1 gene as promoting synergy among one Mot3 site and two Rox1 sites comprising operator A of that gene. Here we studied the role of Mot3 in enhancing repression by Rox1 at another hypoxic gene, HEM13, which is less strongly regulated than ANB1 and has a very different arrangement of Rox1 and Mot3 binding sites. By assessing the effects of deleting Rox1 and Mot3 sites individually and in combination, we found that the major repression of HEM13 occurred through three Mot3 sites closely spaced with a single Rox1 site. While the Mot3 sites functioned additively, they enhanced repression by the single Rox1 site, and the presence of Rox1 enhanced the additive effects of the Mot3 sites. In addition, using a Rox1-Ssn6 fusion protein, we demonstrated that Mot3 enhances Rox1 repression through helping recruit the Ssn6/Tup1 complex. Chromatin immunoprecipitation assays indicated that Rox1 stabilized Mot3 binding to DNA. Integrating these results, we were able to devise a set of rules that govern the combinatorial interactions between Rox1 and Mot3 to achieve differential repression. PMID- 15821126 TI - Functional characterization of an alpha-factor-like Sordaria macrospora peptide pheromone and analysis of its interaction with its cognate receptor in Saccharomyces cerevisiae. AB - The homothallic filamentous ascomycete Sordaria macrospora possesses genes which are thought to encode two pheromone precursors and two seven-transmembrane pheromone receptors. The pheromone precursor genes are termed ppg1 and ppg2. The putative products derived from the gene sequence show structural similarity to the alpha-factor precursors and a-factor precursors of the yeast Saccharomyces cerevisiae. Likewise, sequence similarity has been found between the putative products of the pheromone receptor genes pre2 and pre1 and the S. cerevisiae Ste2p alpha-factor receptor and Ste3p a-factor receptor, respectively. To investigate whether the alpha-factor-like pheromone-receptor pair of S. macrospora is functional, a heterologous yeast assay was used. Our results show that the S. macrospora alpha-factor-like pheromone precursor PPG1 is processed into an active pheromone by yeast MATalpha cells. The S. macrospora PRE2 protein was demonstrated to be a peptide pheromone receptor. In yeast MATa cells lacking the endogenous Ste2p receptor, the S. macrospora PRE2 receptor facilitated all aspects of the pheromone response. Using a synthetic peptide, we can now predict the sequence of one active form of the S. macrospora peptide pheromone. We proved that S. macrospora wild-type strains secrete an active pheromone into the culture medium and that disruption of the ppg1 gene in S. macrospora prevents pheromone production. However, loss of the ppg1 gene does not affect vegetative growth or fertility. Finally, we established the yeast assay as an easy and useful system for analyzing pheromone production in developmental mutants of S. macrospora. PMID- 15821127 TI - Contribution of CAF-I to anaphase-promoting-complex-mediated mitotic chromatin assembly in Saccharomyces cerevisiae. AB - The anaphase-promoting complex (APC) is required for mitotic progression and genomic stability. Recently, we demonstrated that the APC is also required for mitotic chromatin assembly and longevity. Here, we investigated the role the APC plays in chromatin assembly. We show that apc5(CA) mutations genetically interact with the CAF-I genes as well as ASF1, HIR1, and HIR2. When present in multiple copies, the individual CAF-I genes, CAC1, CAC2, and MSI1, suppress apc5(CA) phenotypes in a CAF-1- and Asf1p-independent manner. CAF-I and the APC functionally overlap, as cac1delta cac2delta msi1delta (caf1delta) cells expressing apc5(CA) exhibit a phenotype more severe than that of apc5(CA) or caf1delta. The Ts- phenotypes observed in apc5(CA) and apc5(CA) caf mutants may be rooted in compromised histone metabolism, as coexpression of histones H3 and H4 suppressed the Ts- defects. Synthetic genetic interactions were also observed in apc5(CA) asf1delta cells. Furthermore, increased expression of genes encoding Asf1p, Hir1p, and Hir2p suppressed the apc5(CA) Ts- defect in a CAF-I-dependent manner. Together, these results suggest the existence of a complex molecular mechanism controlling APC-dependent chromatin assembly. Our data suggest the APC functions with the individual CAF-I subunits, Asf1p, and the Hir1p and Hir2p proteins. However, Asf1p and an intact CAF-I complex are dispensable for CAF-I subunit suppression, whereas CAF-I is necessary for ASF1, HIR1, and HIR2 suppression of apc5(CA) phenotypes. We discuss the implications of our observations. PMID- 15821128 TI - Surface localization of the Yps3p protein of Histoplasma capsulatum. AB - The YPS3 gene of Histoplasma capsulatum encodes a protein that is both resident in the cell wall and also released into the culture medium. This protein is produced only during the pathogenic yeast phase of infection and is also expressed differently in H. capsulatum strains that differ in virulence. We investigated the cellular localization of Yps3p. We demonstrated that the cell wall fraction of Yps3p was surface localized in restriction fragment length polymorphism class 2 strains. We also established that Yps3p released into the G217B culture supernatant binds to the surface of strains that do not naturally express the protein. This binding was saturable and occurred within 5 min of exposure and occurred similarly with live and heat-killed H. capsulatum. Flow cytometric analysis of H. capsulatum after enzymatic treatments was consistent with Yps3p binding to chitin, a carbohydrate polymer that is a component of fungal cell walls. Polysaccharide binding assays demonstrated that chitin but not cellulose binds to and extracts Yps3p from culture supernatants. PMID- 15821129 TI - PldB, a putative phospholipase D homologue in Dictyostelium discoideum mediates quorum sensing during development. AB - Quorum sensing, also known as cell-density sensing in the unicellular eukaryote Dictyostelium discoideum, is required for efficient entry into the differentiation and development segment of its life cycle. Quorum sensing is accomplished by simultaneously secreting and sensing the glycoprotein Conditioned Medium Factor, or CMF. When the density of starving cells is high, CMF levels are high, which leads to aggregation followed by development. Here, we describe the role of pldB, a gene coding for a putative phospholipase D (PLD) homologue, in quorum sensing. We find that in submerged culture, adding butanol, an inhibitor of PLD-catalyzed phosphatidic acid production, allows cells to bypass the requirement for CMF mediated quorum sensing and aggregate at low cell density. Deletion of pldB mimics the presence of butanol, allowing cells to aggregate at low cell density. pldB- cells also initiate and finish aggregation rapidly. Analysis of early developmental gene expression in pldB- cells reveals that the cyclic AMP receptor cAR1 is expressed at higher levels earlier than in wild-type cells, which could explain the rapid aggregation phenotype. As would be predicted, cells overexpressing pldB are unable to aggregate even at high cell density. Adding CMF to these pldB- overexpressing cells does not rescue aggregation. Both of these phenotypes are cell autonomous, as mixing a small number of pldB- cells with wild-type cells does not cause the wild-type cells to behave like pldB- cells. PMID- 15821130 TI - Transcriptional response of Saccharomyces cerevisiae to the plasma membrane perturbing compound chitosan. AB - Chitosan is a plasma membrane-perturbing compound consisting of linear chains of beta-1,4-linked glucosamine residues, which at acidic pHs become positively charged. It is extensively used as an antimicrobial compound, yet its mode of action is still unresolved. Chitosan strongly affected the growth of the yeast Saccharomyces cerevisiae, the food spoilage yeast Zygosaccharomyces bailii, and two human-pathogenic yeasts, Candida albicans and Candida glabrata. Microarray analysis of yeast cells treated with sublethal concentrations of chitosan revealed induction of the environmental stress response and three more major transcriptional responses. The first was a rapid and stable Cin5p-mediated response. Cin5p/Yap4p is a transcription factor involved in various stress responses. Deletion of CIN5 led to increased chitosan sensitivity. The second was a Crz1p-mediated response, which is delayed compared to the Cin5p response. Crz1p is a transcription factor of the calcineurin pathway. Cells deleted for CRZ1 or treated with the calcineurin inhibitor FK506 became hypersensitive to chitosan, supporting the notion that the Crz1p-controlled response offers protection against chitosan. The third was a strong Rlm1p-mediated response which ran parallel in time with the Crz1p-regulated response. Rlm1p is a transcription factor of the cell wall integrity pathway, which is activated by cell wall stress. Importantly, chitosan-treated cells became more resistant to beta-1,3 glucanase, which is a well-known response to cell wall stress. We propose that the transcriptional response to chitosan may be representative of other plasma membrane-perturbing compounds. PMID- 15821131 TI - Role of Snf1p in regulation of intracellular sorting of the lactose and galactose transporter Lac12p in Kluyveromyces lactis. AB - The protein kinase Snf1/AMPK plays a central role in carbon and energy homeostasis in yeasts and higher eukaryotes. To work out which aspects of the Snf1-controlled regulatory network are conserved in evolution, the Snf1 requirement in galactose metabolism was analyzed in the yeast Kluyveromyces lactis. Whereas galactose induction was only delayed, K. lactis snf1 mutants failed to accumulate the lactose/galactose H+ symporter Lac12p in the plasma membran,e as indicated by Lac12-green fluorescent protein fusions. In contrast to wild-type cells, the fusion protein was mostly intracellular in the mutant. Growth on galactose and galactose uptake could be restored by the KHT3 gene, which encodes a new transporter of the HXT subfamily of major facilitators These findings indicate a new role of Snf1p in regulation of sugar transport in K. lactis. PMID- 15821132 TI - Identification and gene expression analysis of a large family of transmembrane kinases related to the Gal/GalNAc lectin in Entamoeba histolytica. AB - We identified in the Entamoeba histolytica genome a family of over 80 putative transmembrane kinases (TMKs). The TMK extracellular domains had significant similarity to the intermediate subunit (Igl) of the parasite Gal/GalNAc lectin. The closest homolog to the E. histolytica TMK kinase domain was a cytoplasmic dual-specificity kinase, SplA, from Dictyostelium discoideum. Sequence analysis of the TMK family demonstrated similarities to both serine/threonine and tyrosine kinases. TMK genes from each of six phylogenetic groups were expressed as mRNA in trophozoites, as assessed by spotted oligoarray and real-time PCR assays, suggesting nonredundant functions of the TMK groups for sensing and responding to extracellular stimuli. Additionally, we observed changes in the expression profile of the TMKs in continuous culture. Antisera produced against the conserved kinase domain identified proteins of the expected molecular masses of the expressed TMKs. Confocal microscopy with anti-TMK kinase antibodies revealed a focal distribution of the TMKs on the cytoplasmic face of the trophozoite plasma membrane. We conclude that E. histolytica expresses members of each subgroup of TMKs. The presence of multiple receptor kinases in the plasma membrane offers for the first time a potential explanation of the ability of the parasite to respond to the changing environment of the host. PMID- 15821133 TI - Effect of the major repeat sequence on chromosome loss in Candida albicans. AB - The major repeat sequence (MRS) is found at least once on all but one chromosome in Candida albicans, but as yet it has no known relation to the phenotype. The MRS affects karyotypic variation by serving as a hot spot for chromosome translocation and by expanding and contracting internal repeats, thereby changing chromosome length. Thus, MRSs on different chromosomes and those on chromosome homologues can differ in size. We proposed that the MRS's unique repeat structure and, more specifically, the size of the MRS could also affect karyotypic variation by altering the frequency of mitotic nondisjunction. Subsequent analysis shows that both natural and artificially induced differences in the size of the chromosome 5 MRS can affect chromosome segregation. Strains with chromosome 5 homologues that differ in the size of the naturally occurring MRSs show a preferential loss of the homologue with the larger MRS on sorbose, indicating that a larger MRS leads to a higher risk of mitotic nondisjunction for that homologue. While deletion of an MRS has no deleterious effect on the deletion chromosome under normal growth conditions and leads to no obvious phenotype, strains that have the MRS deleted from one chromosome 5 homologue preferentially lose the homologue with the MRS remaining. This effect on chromosome segregation is the first demonstration of a phenotype associated with the MRS. PMID- 15821134 TI - Structural elements in the 5'-untranslated region of giardiavirus transcript essential for internal ribosome entry site-mediated translation initiation. AB - Translation of uncapped giardiavirus (GLV) mRNA in Giardia lamblia requires the presence of a 5'-untranslated region (5'-UTR) and a viral capsid coding region. We used dicistronic viral constructs to show that the downstream 253 nucleotides (nt) of the 5'-UTR plus the initial 264-nt capsid coding region constitute an internal ribosome entry site (IRES). Predicted secondary structures in the 253-nt 5'-UTR include stem-loops U3, U4a, U4b, U4c, and U5. Chemical and enzymatic probing analysis confirmed the presence of all predicted stem-loops except U4a. Disruption of stem-loop structures U3 and U5 by site-directed mutagenesis resulted in a drastic reduction in translation of a monocistronic viral transcript, which could be restored by compensatory sequence changes. Mutations disrupting stem-loops U4b and U4c do not exert an appreciable effect on translation, but certain sequences in the U4a region and in U4b do appear to play important roles in the IRES. Structural analysis also suggests that an 8-nt U3 loop sequence (nt 147 to 154) pairs with an 8-nt downstream sequence (nt 168 to 175) to form a pseudoknot. Disruption of this pseudoknot by mutagenesis resulted in a drastic reduction in translation, which could be restored by compensatory sequence changes. This study has defined the secondary structure in the 5'-UTR of the IRES. Together with the previous results, we have now completed analysis of the entire structure of GLV IRES and fully defined the functionally essential structural elements in it. PMID- 15821135 TI - Pairwise knockdowns of cdc2-related kinases (CRKs) in Trypanosoma brucei identified the CRKs for G1/S and G2/M transitions and demonstrated distinctive cytokinetic regulations between two developmental stages of the organism. AB - Expression of the cdc2-related kinase 3 (CRK3) together with expression of CRK1, 2, -4, or -6, were knocked down in pairs in the procyclic and bloodstream forms of Trypanosoma brucei, using the RNA interference technique. Double knockdowns of CRK3 and CRK2, CRK4, or CRK6 exerted significant growth inhibition and enriched the cells in G2/M phase, whereas a CRK3 plus CRK1 (CRK3 + CRK1) knockdown arrested cells in both G1/S and G2/M transitions. Thus, CRK1 and CRK3 are apparently the kinases regulating the G1/S and G2/M checkpoint passages, respectively, whereas the other CRKs are probably playing only minor roles in cell cycle regulation. A CRK1 + CRK2 knockdown in the procyclic form was found to cause aberrant posterior cytoskeletal morphogenesis (X. M. Tu and C. C. Wang, Mol. Biol. Cell 16:97-105, 2005). A CRK3 + CRK2 knockdown, however, did not lead to such a change, suggesting that CRK2 depletion can lead to the abnormal morphogenesis only when procyclic-form cells are arrested in the G1 phase. The G2/M-arrested procyclic form produces up to 20% stumpy anucleated cells (zoids) in the population, suggesting that cytokinesis and cell division are not blocked by mitotic arrest but are apparently driven to completion by the kinetoplast cycle. In the bloodstream form, however, G2/M arrest resulted in little zoid formation but, instead, enriched a population of cells each containing multiple kinetoplasts, basal bodies, and flagella and an aggregate of multiple nuclei, indicating failure in entering cytokinesis. The two different cytokinetic regulations between two distinct stage-specific forms of the same organism may provide an interesting and useful model for further understanding the evolution of cytokinetic control among eukaryotes. PMID- 15821136 TI - Mitochondrial DNA ligases of Trypanosoma brucei. AB - The mitochondrial DNA of Trypanosoma brucei, termed kinetoplast DNA or kDNA, consists of thousands of minicircles and a small number of maxicircles catenated into a single network organized as a nucleoprotein disk at the base of the flagellum. Minicircles are replicated free of the network but still contain nicks and gaps after rejoining to the network. Covalent closure of remaining discontinuities in newly replicated minicircles after their rejoining to the network is delayed until all minicircles have been replicated. The DNA ligase involved in this terminal step in minicircle replication has not been identified. A search of kinetoplastid genome databases has identified two putative DNA ligase genes in tandem. These genes (LIG k alpha and LIG k beta) are highly diverged from mitochondrial and nuclear DNA ligase genes of higher eukaryotes. Expression of epitope-tagged versions of these genes shows that both LIG k alpha and LIG k beta are mitochondrial DNA ligases. Epitope-tagged LIG k alpha localizes throughout the kDNA, whereas LIG k beta shows an antipodal localization close to, but not overlapping, that of topoisomerase II, suggesting that these proteins may be contained in distinct structures or protein complexes. Knockdown of the LIG k alpha mRNA by RNA interference led to a cessation of the release of minicircles from the network and resulted in a reduction in size of the kDNA networks and rapid loss of the kDNA from the cell. Closely related pairs of mitochondrial DNA ligase genes were also identified in Leishmania major and Crithidia fasciculata. PMID- 15821137 TI - Intracellular role of adenylyl cyclase in regulation of lateral pseudopod formation during Dictyostelium chemotaxis. AB - Cyclic AMP (cAMP) functions as the extracellular chemoattractant in the aggregation phase of Dictyostelium development. There is some question, however, concerning what role, if any, it plays intracellularly in motility and chemotaxis. To test for such a role, the behavior of null mutants of acaA, the adenylyl cyclase gene that encodes the enzyme responsible for cAMP synthesis during aggregation, was analyzed in buffer and in response to experimentally generated spatial and temporal gradients of extracellular cAMP. acaA- cells were defective in suppressing lateral pseudopods in response to a spatial gradient of cAMP and to an increasing temporal gradient of cAMP. acaA- cells were incapable of chemotaxis in natural waves of cAMP generated by majority control cells in mixed cultures. These results indicate that intracellular cAMP and, hence, adenylyl cyclase play an intracellular role in the chemotactic response. The behavioral defects of acaA- cells were surprisingly similar to those of cells of null mutants of regA, which encodes the intracellular phosphodiesterase that hydrolyzes cAMP and, hence, functions opposite adenylyl cyclase A (ACA). This result is consistent with the hypothesis that ACA and RegA are components of a receptor-regulated intracellular circuit that controls protein kinase A activity. In this model, the suppression of lateral pseudopods in the front of a natural wave depends on a complete circuit. Hence, deletion of any component of the circuit (i.e., RegA or ACA) would result in the same chemotactic defect. PMID- 15821138 TI - A point mutation in the cargo-binding domain of myosin V affects its interaction with multiple cargoes. AB - Class V myosins move diverse intracellular cargoes, which attach via interaction of cargo-specific proteins to the myosin V globular tail. The globular tail of the yeast myosin V, Myo2p, contains two structural and functional subdomains. Subdomain I binds to the vacuole-specific protein, Vac17p, while subdomain II likely binds to an as yet unidentified secretory vesicle-specific protein. All functions of Myo2p require the tight association of subdomains I and II, which suggests that binding of a cargo to one subdomain may inhibit cargo-binding to a second subdomain. Thus, two types of mutations are predicted to specifically affect a subset of Myo2p cargoes: first are mutations within a cargo-specific binding region; second are mutations that mimic the inhibited conformation of one of the subdomains. Here we analyze a point mutation in subdomain I, myo2 2(G1248D), which is likely to be this latter type of mutation. myo2-2 has no effect on secretory vesicle movement. The secretory vesicle binding site is in subdomain II. However, myo2-2 is impaired in several Myo2p-related functions. While subdomains I and II of myo2-2p tightly associate, there are measurable differences in the conformation of its globular tail. Based solely on the ability to restore vacuole inheritance, a set of intragenic suppressors of myo2-2 were identified. All suppressor mutations reside in subdomain I. Moreover, subdomain I and II interactions occurred in all suppressors, demonstrating the importance of subdomain I and II association for Myo2p function. Furthermore, 3 of the 10 suppressors globally restored all tested defects in myo2-2. This large proportion of global suppressors strongly suggests that myo2-2(G1248) causes a conformational change in subdomain I that simultaneously affects multiple cargoes. PMID- 15821139 TI - Systematic deletion analysis of fission yeast protein kinases. AB - Eukaryotic protein kinases are key molecules mediating signal transduction that play a pivotal role in the regulation of various biological processes, including cell cycle progression, cellular morphogenesis, development, and cellular response to environmental changes. A total of 106 eukaryotic protein kinase catalytic-domain-containing proteins have been found in the entire fission yeast genome, 44% (or 64%) of which possess orthologues (or nearest homologues) in humans, based on sequence similarity within catalytic domains. Systematic deletion analysis of all putative protein kinase-encoding genes have revealed that 17 out of 106 were essential for viability, including three previously uncharacterized putative protein kinases. Although the remaining 89 protein kinase mutants were able to form colonies under optimal growth conditions, 46% of the mutants exhibited hypersensitivity to at least 1 of the 17 different stress factors tested. Phenotypic assessment of these mutants allowed us to arrange kinases into functional groups. Based on the results of this assay, we propose also the existence of four major signaling pathways that are involved in the response to 17 stresses tested. Microarray analysis demonstrated a significant correlation between the expression signature and growth phenotype of kinase mutants tested. Our complete microarray data sets are available at http://giscompute.gis.a-star.edu.sg/~gisljh/kinome. PMID- 15821140 TI - Maternal inheritance and stage-specific variation of the apicoplast in Toxoplasma gondii during development in the intermediate and definitive host. AB - The structure and location of Toxoplasma gondii apicoplasts were examined in intermediate and definitive hosts and shown to vary in a stage-specific manner. Immunocytochemistry and electron microscopy studies were used to identify changes in the morphology of apicoplasts and in their enoyl reductase (ENR) content during asexual and sexual development. Apicoplasts in tachyzoites were small, multimembraned organelles anterior to nuclei that divided and segregated with the nuclei during endodyogeny. In nonproliferating bradyzoites within mature tissue cysts (1 to 24 months), apicoplasts had high levels of ENR. During coccidian development, asexual multiplication (endopolygeny), resulting in simultaneous formation of up to 30 daughters (merozoites), involved an initial growth phase associated with repeated nuclear divisions during which apicoplasts appeared as single, elongated, branched structures with increased levels of ENR. At initiation of merozoite formation, enlarged apicoplasts divided simultaneously, with constrictions, into portions that segregated to developing daughters. In sexual stages, apicoplast division did not occur during microgametogony, and apicoplasts were absent from the microgametes that were formed. In contrast, during macrogametogony, the apicoplast appeared as a large, branched, perinuclear structure that had very high levels of ENR in the absence of nuclear division. Marked increases in the size of apicoplasts and levels of ENR may be related to requirements of the macrogametocytes to synthesize and store all components necessary for oocyst formation and subsequent extracellular sporulation. Thus, it is shown that apicoplasts are present and contain ENR in all T. gondii life cycle stages except microgametes, which will result in maternal inheritance of the organelle. PMID- 15821142 TI - The highly conserved tRNAHis guanylyltransferase Thg1p interacts with the origin recognition complex and is required for the G2/M phase transition in the yeast Saccharomyces cerevisiae. AB - Here we show that the Saccharomyces cerevisiae tRNA(His) guanylyltransferase Thg1p interacts with the origin recognition complex in vivo and in vitro and that overexpression of hemagglutinin-Thg1p selectively impedes growth of orc2-1(Ts) cells at the permissive temperature. Studies with conditional mutants indicate that Thg1p couples nuclear division and migration to cell budding and cytokinesis in yeast. PMID- 15821141 TI - Proteomic analysis of phagocytosis in the enteric protozoan parasite Entamoeba histolytica. AB - Proteomic analysis of phagosomes isolated from Entamoeba histolytica by liquid chromatography and mass spectrometry identified 85 proteins involved in surface recognition, actin cytoskeleton rearrangement, vesicular trafficking, and degradation. Phagosome localization of representative proteins was verified by immunofluorescence assay. This study should provide a basis for molecular identification and characterization of phagosome biogenesis. PMID- 15821143 TI - A plant notices insect egg deposition and changes its rate of photosynthesis. AB - Scots pine (Pinus sylvestris) is known to change its terpenoid metabolism in response to egg deposition by the sawfly Diprion pini (Hymenoptera, Diprionidae). Three days after egg deposition, parts of the pine twig adjacent to the egg-laden one are induced to emit volatiles, which attract egg parasitoids. In this study, we investigated whether egg deposition by this sawfly affects pine photosynthesis. Measurements of photosynthesis were taken from untreated control twigs and from pine twigs adjacent to egg-laden ones (i.e. systemically oviposition-induced twigs) for a period of 3 d starting after egg deposition. The net photosynthetic rate of oviposition-induced pine twigs was lower than that of untreated control twigs, whereas the respiration rate of pine twigs was not affected by egg deposition. CO2 response curves of oviposition-induced twigs tended to be lower than those of controls. The potential rate of electron transport (J(max)) and the maximum rate of Rubisco activity (V(cmax)) were calculated from the data of the CO2 response curves. J(max) of oviposition induced twigs was significantly lower than that of controls at day 1 after egg deposition, while the difference diminished from day 2 to day 3. A similar pattern was observed for V(cmax). Light response curves of oviposition-induced twigs were significantly lower than those of untreated ones during 3 d of measurements. Stomatal conductance was slightly lowered by egg deposition. When considering photosynthetic activity as a physiological currency to measure costs of induction of plant defense, the effects of insect egg deposition on gas exchange of pine are discussed with respect to known effects of insect feeding on the photosynthesis activity of plants. PMID- 15821144 TI - A stilbene synthase gene (SbSTS1) is involved in host and nonhost defense responses in sorghum. AB - A chalcone synthase (CHS)-like gene, SbCHS8, with high expressed sequence tag abundance in a pathogen-induced cDNA library, was identified previously in sorghum (Sorghum bicolor). Genomic Southern analysis revealed that SbCHS8 represents a single-copy gene. SbCHS8 expression was induced in sorghum mesocotyls following inoculation with Cochliobolus heterotrophus and Colletotrichum sublineolum, corresponding to nonhost and host defense responses, respectively. However, the induction was delayed by approximately 24 h when compared to the expression of at least one of the other SbCHS genes. In addition, SbCHS8 expression was not induced by light and did not occur in a tissue-specific manner. SbCHS8, together with SbCHS2, was overexpressed in transgenic Arabidopsis (Arabidopsis thaliana) tt4 (transparent testa) mutants defective in CHS activities. SbCHS2 rescued the ability of these mutants to accumulate flavonoids in seed coats and seedlings. In contrast, SbCHS8 failed to complement the mutation, suggesting that the encoded enzyme does not function as a CHS. To elucidate their biochemical functions, recombinant proteins were assayed with different phenylpropanoid-Coenzyme A esters. Flavanones and stilbenes were detected in the reaction products of SbCHS2 and SbCHS8, respectively. Taken together, our data demonstrated that SbCHS2 encodes a typical CHS that synthesizes naringenin chalcone, which is necessary for the formation of different flavonoid metabolites. On the other hand, SbCHS8, now retermed SbSTS1, encodes an enzyme with stilbene synthase activity, suggesting that sorghum accumulates stilbene-derived defense metabolites in addition to the well characterized 3-deoxyanthocyanidin phytoalexins. PMID- 15821145 TI - Why is golden rice golden (yellow) instead of red? AB - The endosperm of Golden Rice (Oryza sativa) is yellow due to the accumulation of beta-carotene (provitamin A) and xanthophylls. The product of the two carotenoid biosynthesis transgenes used in Golden Rice, phytoene synthase (PSY) and the bacterial carotene desaturase (CRTI), is lycopene, which has a red color. The absence of lycopene in Golden Rice shows that the pathway proceeds beyond the transgenic end point and thus that the endogenous pathway must also be acting. By using TaqMan real-time PCR, we show in wild-type rice endosperm the mRNA expression of the relevant carotenoid biosynthetic enzymes encoding phytoene desaturase, zeta-carotene desaturase, carotene cis-trans-isomerase, beta-lycopene cyclase, and beta-carotene hydroxylase; only PSY mRNA was virtually absent. We show that the transgenic phenotype is not due to up-regulation of expression of the endogenous rice pathway in response to the transgenes, as was suggested to be the case in tomato (Lycopersicon esculentum) fruit, where CRTI expression resulted in a similar carotenoid phenomenon. This means that beta-carotene and xanthophyll formation in Golden Rice relies on the activity of constitutively expressed intrinsic rice genes (carotene cis-trans-isomerase, alpha/beta-lycopene cyclase, beta-carotene hydroxylase). PSY needs to be supplemented and the need for the CrtI transgene in Golden Rice is presumably due to insufficient activity of the phytoene desaturase and/or zeta-carotene desaturase enzyme in endosperm. The effect of CRTI expression was also investigated in leaves of transgenic rice and Arabidopsis (Arabidopsis thaliana). Here, again, the mRNA levels of intrinsic carotenogenic enzymes remained unaffected; nevertheless, the carotenoid pattern changed, showing a decrease in lutein, while the beta-carotene-derived xanthophylls increased. This shift correlated with CRTI-expression and is most likely governed at the enzyme level by lycopene-cis-trans-isomerism. Possible implications are discussed. PMID- 15821146 TI - The pivotal roles of the plant S-adenosylmethionine decarboxylase 5' untranslated leader sequence in regulation of gene expression at the transcriptional and posttranscriptional levels. AB - S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.1.50) is a key rate-limiting enzyme located in the polyamine biosynthesis pathway. When compared with other organisms, the plant SAMDC genes possess some distinct features because they are devoid of introns in the main open reading frame (ORF) but have an intron(s) in their 5' untranslated leader sequences, in which two overlapping tiny and small upstream ORFs (uORFs) are present. Our results show that the presence of the 5' leader sequence plays important roles in transcriptional and posttranscriptional regulation of SAMDC expression. This sequence may help to keep the transcript of its downstream cistron at a relatively low level and function together with its own promoter in response to external stimuli or internal changes of spermidine and spermine to initiate and regulate SAMDC expression. Under stress and high spermidine or spermine conditions, the tiny uORF shows the same function as its overlapping small uORF, which is involved in translational repression and feedback controlled by polyamines. The presence of introns is necessary for the SAMDC up-regulation process when the internal spermidine level is low. Our results suggest that plants have evolved one network to adjust SAMDC activity through their 5' leader sequences, through which transcriptional regulation is combined with an extensive posttranscriptional control circuit. PMID- 15821147 TI - Molecular cloning of GA 2-oxidase3 from spinach and its ectopic expression in Nicotiana sylvestris. AB - Previous work has shown that 13-hydroxylated gibberellins (GAs) are predominant in the long-day (LD) plant spinach (Spinacia oleracea; GA53, GA44, GA19, GA20, GA1, GA8, and GA29). Also present in spinach are 2-hydroxylated C20-GAs: GA97, GA98, GA99, and GA110. Levels of the most abundant GA, GA97, decreased when plants were transferred from short photoperiods (SD) to LD. When [14C]GA53 was fed to spinach plants, more GA53 was converted to GA97 in SD than in LD, and more radioactive GA20 was formed in LD than in SD. SoGA2ox3, encoding a GA 2-oxidase, was isolated from spinach. The recombinant protein converted only two C20-GA precursors, GA12 and GA53, to their respective products, GA110 and GA97. GA2ox3 competes with GA20ox1 for their common substrate, GA53. In SD, deactivation to GA97 prevails, whereas in LD conversion to GA20 is favored. Transcript levels of SoGA2ox3 were higher in shoot tips than in blades, petioles, and young leaves. Ectopic expression of SoGA2ox3 in the long-day plant Nicotiana sylvestris showed a range of dwarf phenotypes, such as reduced germination, short hypocotyl and stem, dark-green leaves, and late flowering, but normal flowers and seed production. The levels of GA53 and GA1 were 3- to 5-fold lower in transgenic plants than in wild type, whereas the levels of GA97 and GA110 increased 3- to 6 fold in transgenic plants. It is concluded that genetic manipulation of plant stature by increasing deactivation of precursors of active GA is more advantageous than increased deactivation of bioactive GA1 itself. PMID- 15821148 TI - STIG1 controls exudate secretion in the pistil of petunia and tobacco. AB - The lipid-rich, sticky exudate covering the stigma of solanaceous species such as tobacco (Nicotiana tabacum) and petunia (Petunia hybrida) contains several proteins, of which only some have been characterized to date. Proteome analysis of the stigmatic exudate in both species revealed the presence of a cysteine rich, slightly acidic 12-kD protein called stigma-specific protein 1 (STIG1). In both tobacco and petunia, Stig1 is highly expressed at the mRNA level in very young and developing flowers, whereas hardly any Stig1 transcript is detected in mature flowers. This expression pattern coincides with the differentiation of the secretory zone, forming the intercellular spaces into which the exudate is secreted. Using reverse genetics, we show that STIG1 is involved in the secretion and merging of exudate lipids in the intercellular spaces of the secretory zone and that plants lacking STIG1 show an accelerated deposition of exudate onto the stigmatic surface. This phenotype was observed both in a petunia knockout mutant and in tobacco transgenic plants. We therefore propose that STIG1 plays a role in the temporal regulation of the essential exudate secretion onto the stigma. PMID- 15821150 TI - Nanomolar and micromolar effects of 15-deoxy-delta 12,14-prostaglandin J2 on amnion-derived WISH epithelial cells: differential roles of peroxisome proliferator-activated receptors gamma and delta and nuclear factor kappa B. AB - 15-Deoxy delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), an activator of peroxisome proliferator-activated receptor (PPAR)-gamma and -delta, is a prostanoid metabolite with anti-inflammatory actions. In intrauterine tissues, proinflammatory cytokines and prostaglandins have been identified as playing key roles in the maintenance of pregnancy and the onset of labor. We investigated and compared the early (<3 h) effects of 15d-PGJ(2) with rosiglitazone (PPAR-gamma ligand) and 2-methyl-4-((4-methyl-2-(4-trifluoromethylphenyl)-1,3-thiazol-5-yl) methylsulfanyl)phenoxy-acetic acid (GW501516) (PPAR-delta ligand) on interleukin (IL)-1beta-induced prostaglandin and cytokine production by amnion-derived WISH cells. We show that 15d-PGJ(2) exerts differential effects depending on concentration. At low concentrations (<0.1 microM), 15d-PGJ(2) inhibited IL-1beta stimulated prostaglandin E(2) (PGE(2)) but not cytokine (IL-6/IL-8) production or cyclooxygenase-2 (COX-2) expression. This effect was attenuated by a PPAR-gamma inhibitor [2-chloro-5-nitro-N-phenyl-benzamide (GW9662)], by transfection with a dominant-negative PPAR construct, and was reproduced by the PPAR-gamma ligand rosiglitazone. At higher concentrations (1-10 microM), 15d-PGJ(2) inhibited IL 1beta-stimulated PGE(2) and cytokine production and COX-2 expression, and this effect was not blocked by GW9662. Rosiglitazone at high concentrations (1-10 microM) stimulated PGE(2) production in the absence or presence of the dominant negative PPAR. The PPAR-delta ligand GW501516 also inhibited IL-1beta-stimulated PGE(2) production but only at high concentrations (1 microM). IL-1beta-induced nuclear factor-kappaB (NF-kappaB) DNA binding activity was significantly inhibited by 15d-PGJ(2) (10 microM) and GW501516 (1 microM) but increased with 10 microM rosiglitazone. We conclude that 1) at low concentrations, 15d-PGJ(2) acts through a PPAR-gamma signaling pathway; b) at higher concentrations, its actions are mediated most likely through other pathways such as activation of PPAR-delta and/or inhibition of NF-kappaB; and 3) rosiglitazone exerts PPAR-independent effects at high concentrations (>1 microM). PMID- 15821149 TI - Identification of a long-chain polyunsaturated fatty acid acyl-coenzyme A synthetase from the diatom Thalassiosira pseudonana. AB - The draft genome of the diatom Thalassiosira pseudonana was searched for DNA sequences showing homology with long-chain acyl-coenzyme A synthetases (LACSs), since the corresponding enzyme may play a key role in the accumulation of health beneficial polyunsaturated fatty acids (PUFAs) in triacylglycerol. Among the candidate genes identified, an open reading frame named TplacsA was found to be full length and constitutively expressed during cell cultivation. The predicted amino acid sequence of the corresponding protein, TpLACSA, exhibited typical features of acyl-coenzyme A (acyl-CoA) synthetases involved in the activation of long-chain fatty acids. Feeding experiments carried out in yeast (Saccharomyces cerevisiae) transformed with the algal gene showed that TpLACSA was able to activate a number of PUFAs, including eicosapentaenoic acid and docosahexaenoic acid (DHA). Determination of acyl-CoA synthetase activities by direct measurement of acyl-CoAs produced in the presence of different PUFA substrates showed that TpLACSA was most active toward DHA. Heterologous expression also revealed that TplacsA transformants were able to incorporate more DHA in triacylglycerols than the control yeast. PMID- 15821152 TI - The duplicitous nature of inorganic arsenic. PMID- 15821153 TI - Eric Kandel: the future of memory. AB - Watching ice floes glide by on the Hudson River from Eric Kandel's office, one gets a sense of placid reflection tempered by constant action-an apt analogy for Kandel's ability to calmly manage several ongoing projects and commitments at once. In addition to his well-lauded, ongoing research at Columbia University Medical Center's New York State Psychiatric Institute, Kandel has written several books on neurobiology, behavior, and memory. In addition to being a Nobel Laureate Scientist, he is well-known as an editor of the seminal textbook Principles of Neural Science. He and his colleagues are in the midst of working on a new edition of Principles, and he is working on a scientific autobiography. MI sat down with Dr. Kandel and discussed with him a range of topics including childhood and early career influences, intramural research at the NIH, the HHMI, ethical considerations of altering memory and, of course, Aplysia. PMID- 15821154 TI - NAADP: a new second messenger comes of age. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) is one of the most potent stimulators of intracellular Ca(2+) mobilization in a variety of cell types. Its role in physiological processes is increasingly demonstrated by NAADP increases following cellular stimulation. As a second messenger NAADP shows unique features such as the ability to mobilize Ca(2+) from stores that are physically distinct from those connected to the Ca(2+) channels located in the endoplasmic reticulum, namely, the inositol-1,4,5-trisphosphate and the cyclic-ADP-ribose/ ryanodine receptors. Furthermore, the NAADP-induced self-inactivation mechanism is suggestive of an irreversible binding of NAADP to its putative receptor. PMID- 15821155 TI - The NAADP receptor: new receptors or new regulation? AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) is the most potent calcium mobilizing messenger yet discovered. Its action has now been reported in a large number of cell types from a diverse array of organisms, and in some cases linked to the transduction of specific cellular stimuli. However, what is controversial is the nature of its target calcium release channel, as well as the subcellular localization of its receptor. Some have proposed that NAADP activates a novel calcium release channel distinct from the two major classes of channels known, the inositol trisphosphate receptors and ryanodine receptors. However, others have suggested that it acts in a novel way to regulate a known calcium release channel, the ryanodine receptor. PMID- 15821156 TI - There are no useful CYP3A probes that quantitatively predict the in vivo kinetics of other CYP3A substrates and no expectation that one will be found. AB - The search for a substrate that may serve as a probe to quantitatively predict the in vivo kinetics of cytochrome P450 3A (CYP3A) drugs has been of particular interest because more than half of all human drugs appear to be substrates for this enzyme. Even three closely related 1,4-benzodiazepines-alprazolam, midazolam, and triazolam-are inadequate probes to predict the pharmacokinetics of each other in an individual. If these drugs--all metabolized through the same CYP3A pathways in humans, all FDA Biopharmaceutical Classification System Class 1 compounds exhibiting high solubility and high permeability and thus unaffected by transporter differences--cannot quantitatively predict the pharmacokinetics of their closely related congeners, there is little hope that any quantitative CYP3A probe will ever be found. PMID- 15821157 TI - Mechanism of tissue-selective drug action in the cardiovascular system. AB - Analysis of the human genome project tells us that there may be as few as 3000 genes that are likely to be good drug targets. Although the number of targets is still very large, these data have been interpreted by some to mean that the pharmaceutical industry may someday run out of novel drug targets. Despite the doom and gloom of such analysis, there is considerable reason for optimism. Drugs may exhibit selectivity of action beyond that predicted by target expression alone. Drugs that act at a single molecular target may have very different pharmacology and, as a result, different therapeutic uses. Three well characterized model systems are highlighted to illustrate this point. The first model system is exemplified by nifedipine and verapamil, both of which act on L type calcium channels. Both drugs are used to treat hypertension, but only verapamil can be used to produce atrioventricular block in patients with atrial fibrillation. The second model system describes the therapeutic exploitation of unusual conditions that occur in the ischemic myocardium to produce drugs that are more effective for suppressing ischemia-induced arrhythmias. The third model system discusses the mechanisms through which phosphodiesterase-5 (PDE5) inhibitors act selectively to facilitate penile erection while having little effect in the non-penile vasculature that also expresses PDE5. PMID- 15821158 TI - Role of mitochondria in toxic oxidative stress. AB - Oxidative stress and mitochondrial oxidative damage have been implicated in the etiology of numerous common diseases. The critical mitochondrial events responsible for oxidative stress-mediated cell death (toxic oxidative stress), however, have yet to be defined. Several oxidative events implicated in toxic oxidative stress include alterations in mitochondrial lipids (e.g., cardiolipin), mitochondrial DNA, and mitochondrial proteins (eg. aconitase and uncoupling protein 2). Furthermore, recent findings indicate the enrichment of mitochondrial membranes with vitamin E protects cells against the toxic effects of oxidative stress. This review briefly summarizes the role of these mitochondrial events in toxic oxidative stress, including: 1) the protective role of mitochondrial vitamin E in toxic oxidative stress, 2) the role of mitochondrial DNA in toxic oxidative stress, 3) the interaction between cardiolipin and cytochrome c in mitochondrial regulation of apoptosis, 4) the role of mitochondrial aconitase in oxidative neurodegeneration, and 5) the role of mitochondrial uncoupling protein 2 in the pathogenesis of type 2 diabetes. PMID- 15821160 TI - Solid phase DNA amplification: a Brownian dynamics study of crowding effects. AB - Solid phase amplification (SPA), a new method to amplify DNA, is characterized by the use of surface-bound primers. This limits the amplification to two dimensional surfaces and therefore allows the easy parallelization of DNA amplification in a single system. SPA leads to the formation of small but dense DNA brushes, called DNA colonies. For a molecule to successfully duplicate itself, it needs to bend so that its free end can find a matching primer, located on the surface. We used Brownian dynamics simulations (with a united-atom model) to model the basic kinetics of an SPA experiment. The simulations mimic the temperature cycles and the molecule duplication process found in SPA. Our results indicate that the steric interaction between molecules leads to a decreased duplication probability for molecules in the center of a colony and to an outward leaning for the molecules on the perimeter. These effects result in slower amplification (compared to solution PCR) and indicate that steric interaction alone can explain the loss of the exponential growth (characteristic of solution PCR) of the number of molecules in an SPA experiment. Furthermore, the growth of the colony as a function of the number of thermal cycles is found to be similar to the one obtained with a simple Monte Carlo simulation. PMID- 15821159 TI - Twenty years of calcium imaging: cell physiology to dye for. AB - The use of fluorescent dyes over the past two decades has led to a revolution in our understanding of calcium signaling. Given the ubiquitous role of Ca(2+) in signal transduction at the most fundamental levels of molecular, cellular, and organismal biology, it has been challenging to understand how the specificity and versatility of Ca(2+) signaling is accomplished. In excitable cells, the coordination of changing Ca(2+) concentrations at global (cellular) and well defined subcellular spaces through the course of membrane depolarization can now be conceptualized in the context of disease processes such as cardiac arrhythmogenesis. The spatial and temporal dimensions of Ca(2+) signaling are similarly important in non-excitable cells, such as endothelial and epithelial cells, to regulate multiple signaling pathways that participate in organ homeostasis as well as cellular organization and essential secretory processes. PMID- 15821161 TI - Protein structural deformation induced lifetime shortening of photosynthetic bacteria light-harvesting complex LH2 excited state. AB - Photosynthetic bacterial light-harvesting antenna complex LH2 was immobilized on the surface of TiO(2) nanoparticles in the colloidal solution. The LH2/TiO(2) assembly was investigated by the time-resolved spectroscopic methods. The excited state lifetimes for carotenoid-containing and carotenoidless LH2 have been measured, showing a decrease in the excited-state lifetime of B850 when LH2 was immobilized on TiO(2). The possibility that the decrease of the LH2 excited-state lifetime being caused by an interfacial electron transfer reaction between B850 and the TiO(2) nanoparticle was precluded experimentally. We proposed that the observed change in the photophysical properties of LH2 when assembled onto TiO(2) nanoparticles is arising from the interfacial-interaction-induced structural deformation of the LH2 complex deviating from an ellipse of less eccentric to a more eccentric ellipse, and the observed phenomenon can be accounted by an elliptical exciton model. Experiment by using photoinactive SiO(2) nanoparticle in place of TiO(2) and core complex LH1 instead of LH2 provide further evidence to the proposed mechanism. PMID- 15821162 TI - Cold instability of aponeocarzinostatin and its stabilization by labile chromophore. AB - The conformational stability of aponeocarzinostatin, an all-beta-sheet protein with 113 amino-acid residues, is investigated by thermal-induced equilibrium unfolding between pH 2.0 and 10.0 with and without urea. At room temperature, the protein is stable in a pH range of 4.0-10.0, whereas the stability of the protein drastically decreases below pH 4.0. The thermal unfolding of aponeocarzinostatin is reversible and follows a two-state mechanism. By two-dimensional unfolding studies, the enthalpy change, heat capacity change, and free energy change for unfolding of the protein are estimated. Circular dichroism profiles suggest that this protein undergoes both heat- and cold-induced unfolding. The ellipticity changes at far- and near-UV circular dichroism suggest that the tertiary structure is disrupted but the secondary structure remains folded at low temperatures. Interestingly, the labile enediyne chromophore, which is highly stabilized by the protein, is able to protect the protein against cold-induced unfolding, but not the heat-induced unfolding. PMID- 15821163 TI - Proton binding to proteins: a free-energy component analysis using a dielectric continuum model. AB - Proton binding plays a critical role in protein structure and function. We report pK(a) calculations for three aspartates in two proteins, using a linear response approach, as well as a "standard" Poisson-Boltzmann approach. Averaging over conformations from the two endpoints of the proton-binding reaction, the protein's atomic degrees of freedom are explicitly modeled. Treating macroscopically the protein's electronic polarizability and the solvent, a meaningful model is obtained, without adjustable parameters. It reproduces qualitatively the electrostatic potentials, proton-binding free energies, Marcus reorganization free energies, and pK(a) shifts from explicit solvent molecular dynamics simulations, and the pK(a) shifts from experiment. For thioredoxin Asp 26, which has a large pK(a) upshift, we correctly capture the balance between unfavorable carboxylate desolvation and favorable interactions with a nearby lysine; similarly for RNase A Asp-14, which has a large pK(a) downshift. For the unshifted thioredoxin Asp-20, desolvation by the protein cavity is overestimated by 2.9 pK(a) units; several effects could explain this. "Standard" Poisson Boltzmann methods sidestep this problem by using a large, ad hoc protein dielectric; but protein charge-charge interactions are then incorrectly downscaled, giving an unbalanced description of the reaction and a large error for the shifted pK(a) values of Asp-26 and Asp-14. PMID- 15821164 TI - Functional dynamics of PDZ binding domains: a normal-mode analysis. AB - Postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains are relatively small (80-120 residues) protein binding modules central in the organization of receptor clusters and in the association of cellular proteins. Their main function is to bind C-terminals of selected proteins that are recognized through specific amino acids in their carboxyl end. Binding is associated with a deformation of the PDZ native structure and is responsible for dynamical changes in regions not in direct contact with the target. We investigate how this deformation is related to the harmonic dynamics of the PDZ structure and show that one low-frequency collective normal mode, characterized by the concerted movements of different secondary structures, is involved in the binding process. Our results suggest that even minimal structural changes are responsible for communication between distant regions of the protein, in agreement with recent NMR experiments. Thus, PDZ domains are a very clear example of how collective normal modes are able to characterize the relation between function and dynamics of proteins, and to provide indications on the precursors of binding/unbinding events. PMID- 15821165 TI - Selective field effects on intracellular vacuoles and vesicle membranes with nanosecond electric pulses. AB - Electric pulses across intact vesicles and cells can lead to transient increase in permeability of their membranes. We studied the integrity of these membranes in response to external electric pulses of high amplitude and submicrosecond duration with a primary aim of achieving selective permeabilization. These effects were examined in two separate model systems comprising of 1), a mixed population of 1,2-di-oleoyl-sn-glycero-3-phosphocholine phospholipid vesicles and in 2), single COS-7 cells, in which large endosomal membrane vacuoles were induced by stimulated endocytosis. It has been shown that large and rapidly varying external electric fields, with pulses shorter than the charging time of the outer-cell membrane, could substantially increase intracellular fields to achieve selective manipulations of intracellular organelles. The underlying principle of this earlier work is further developed and applied to the systems studied here. Under appropriate conditions, we show preferential permeabilization of one vesicle population in a mixed preparation of vesicles of similar size distribution. It is further shown that large endocytosed vacuoles in COS-7 cells can be selectively permeabilized with little effect on the integrity of outer cell membrane. PMID- 15821167 TI - High spatial resolution observation of single-molecule dynamics in living cell membranes. AB - Self-organized lipid bilayers together with proteins are the essential building blocks of biological membranes. Membranes are associated with all living systems as they make up cell boundaries and provide basic barriers to cellular organelles. It is of interest to study the dynamics of individual molecules in cell membranes as the mechanism of how biological membranes function at the single molecule remains to be elucidated. In this letter we describe a study in which we incubate rat basophilic leukemia cells with a fluorescently labeled cell membrane component on a surface containing zero-mode waveguides (ZMWs). We used the ZMW to confine fluorescent excitation to an approximately 100-nm region of the membrane to monitor lipid diffusion along the cellular membrane. We showed that confinement with a ZMW largely reduced fluorescent contributions from the cytosolic pool that is present when using a more standard technique such as laser induced confocal microscopy. We show that optical confinement with ZMWs is a facile way to probe dynamic processes on the membrane surface. PMID- 15821166 TI - Single-molecule studies of synaptotagmin and complexin binding to the SNARE complex. AB - The assembly of multiprotein complexes at the membrane interface governs many signaling processes in cells. However, very few methods exist for obtaining biophysical information about protein complex formation at the membrane. We used single molecule fluorescence resonance energy transfer to study complexin and synaptotagmin interactions with the SNARE complex in deposited lipid bilayers. Using total internal reflectance microscopy, individual binding events at the membrane could be resolved despite an excess of unbound protein in solution. Fluorescence resonance energy transfer (FRET)-efficiency derived distances for the complexin-SNARE interaction were consistent with the crystal structure of the complexin-SNARE complex. The unstructured N-terminal region of complexin showed broad distributions of FRET efficiencies to the SNARE complex, suggesting that information on conformational variability can be obtained from FRET efficiency distributions. The low-affinity interaction of synaptotagmin with the SNARE complex changed dramatically upon addition of Ca2+ with high FRET efficiency interactions appearing between the C2B domain and linker domains of synaptotagmin and the membrane proximal portion of the SNARE complex. These results demonstrate that single molecule FRET can be used as a "spectroscopic ruler" to simultaneously gain structural and kinetic information about transient multiprotein complexes at the membrane interface. PMID- 15821168 TI - Loop conformation and dynamics of the Escherichia coli HPPK apo-enzyme and its binary complex with MgATP. AB - Comparison of the crystallographic and NMR structures of 6-hydroxymethyl-7,8 dihydropterin pyrophosphokinase (HPPK) suggests that the enzyme may undergo significant conformational change upon binding to its first substrate, ATP. Two of the three surface loops (loop 2 and loop 3) accounting for most of the conformational differences appear to be confined by crystal contacts, raising questions about the putative large-scale induced-fit conformational change of HPPK and the functional roles of the conserved side-chain residues on the loops. To investigate the loop dynamics in crystal-free environment, we carried out molecular dynamics and locally enhanced sampling simulations of the apo-enzyme and the HPPK.MgATP complex. Our simulations showed that the crystallographic B factors underestimated the loop dynamics considerably. We found that the open conformation of loop 3 in the binary complex is accessible to the apo-enzyme and is the favored conformation in solution phase. These results revise our previous view of HPPK-substrate interactions and the associated functional mechanism of conformational change. The lessons learned here offer valuable structural insights into the workings of HPPK and should be useful for structure-based drug design. PMID- 15821169 TI - The protonation-deprotonation kinetics of the protonated Schiff base in bicelle bacteriorhodopsin crystals. AB - In the recently published x-ray crystal structure of the "bicelle" bacteriorhodopsin (bbR) crystal, the protein has quite a different structure from the native and the in cubo bacteriorhodopsin (cbR) crystal. Instead of packing in parallel trimers as do the native membrane and the cbR crystals, in the bbR crystal the protein packs as antiparallel monomers. To date, no functional studies have been performed, to our knowledge, to investigate if the photocycle is observed in this novel protein packing structure. In this study, both Raman and time-resolved transient absorption spectroscopy are used to both confirm the presence of the photocycle and investigate the deprotonation-reprotonation kinetics of the Schiff base proton in the bbR crystal. The observed rates of deprotonation and reprotonation processes of its Schiff base have been compared to those observed for native bR under the same conditions. Unlike the previously observed similarity of the rates of these processes for cbR crystals and those for native bacteriorhodopsin (bR), in bbR crystals the rate of deprotonation has increased by 300%, and the rate of reprotonation has decreased by nearly 700%. These results are discussed in light of the changes observed when native bR is delipidated or monomerized by detergents. Both the change of the hydrophobicity of the environment around the protonated Schiff base and Asp85 and Asp96 (which could change the pKa values of proton donor-acceptor pairs) and the water structure in the bbR crystal are offered as possible explanations for the different observations. PMID- 15821170 TI - Initial conformational changes of human transthyretin under partially denaturing conditions. AB - Human transthyretin (TTR) is an amyloidogenic protein. The pathway of TTR amyloid formation has been proposed based on lines of evidence: TTR tetramer first dissociates into native monomers, which is shown to be a rate-limiting step in the formation of fibrils. Subsequently, the monomeric species partially unfold to form the aggregation intermediates. Once such intermediates are formed, the following self-assembly process is a downhill polymerization. Hence, tertiary structural changes within the monomers after the dissociation are essential for the amyloid formation. These tertiary structural changes can be facilitated by partial denaturation. To probe the conformational changes under the partially denaturing conditions, five independent trajectories were collected for the wild type (WT) and its pathogenic variants at 300 and 350 K, resulting in simulations that totaled 59 ns. Under these conditions, L55P variant is more labile than the wild-type and V30M variant. We have observed that the D strand of WT-TTR is trapped in two local minima: the native conformation and the amyloidogenic fold that resembles the surface loop of residues 54-55 of L55P variant. In the tetrameric state, the F strand is bent with large separations at the F-F' interface. This strand becomes flatter in the monomeric state, which may facilitate the formation of new F-F' interface with possible prolonged hydrogen bonds and/or shift in beta-strand register in the fibril state. During the unfolding process, the anticorrelated motion between the strands H and G as well as the strands H and A pulls the H strand out of the inner sheet plane, leading to a more twisted inner sheet. Our simulation has provided important detailed structural information about the partially unfolded state of TTR that may be related to the amyloidogenic intermediates. PMID- 15821171 TI - Epithelial Na+ channel subunit stoichiometry. AB - Ion channels, including the epithelial Na(+) channel (ENaC), are intrinsic membrane proteins comprised of component subunits. Proper subunit assembly and stoichiometry are essential for normal physiological function of the channel protein. ENaC comprises three subunits, alpha, beta, and gamma, that have common tertiary structures and much amino acid sequence identity. For maximal ENaC activity, each subunit is required. The subunit stoichiometry of functional ENaC within the membrane remains uncertain. We combined a biophysical approach, fluorescence intensity ratio analysis, used to assess relative subunit stoichiometry with total internal reflection fluorescence microscopy, which enables isolation of plasma membrane fluorescence signals, to determine the limiting subunit stoichiometry of ENaC within the plasma membrane. Our results demonstrate that membrane ENaC contains equal numbers of each type of subunit and that at steady state, subunit stoichiometry is fixed. Moreover, we find that when all three ENaC subunits are coexpressed, heteromeric channel formation is favored over homomeric channels. Electrophysiological results testing effects of ENaC subunit dose on channel activity were consistent with total internal reflection fluorescence/fluorescence intensity ratio findings and confirmed preferential formation of heteromeric channels containing equal numbers of each subunit. PMID- 15821172 TI - Varying approaches to tracheostomy: "vive la difference". PMID- 15821173 TI - Pulmonary and critical care: the unattractive specialty. PMID- 15821174 TI - End of the idyll: farming and the risk of occupational allergy. PMID- 15821175 TI - Autonomic function, omega-3, and cardiovascular risk. PMID- 15821176 TI - To screen or not to screen. PMID- 15821177 TI - The more, the better: maximum oxygen uptake and lung resection. PMID- 15821178 TI - Integrating survival and quality of life data in clinical trials of lung disease: the case of lung volume reduction surgery. PMID- 15821179 TI - Proton-pump inhibitor therapy for gastroesophageal reflux disease: does it treat the asthma? PMID- 15821180 TI - Inhaled tobramycin: not just for cystic fibrosis anymore? PMID- 15821181 TI - Cardiac autonomic changes associated with fish oil vs soy oil supplementation in the elderly. AB - INTRODUCTION: Omega-3 fatty acid levels are associated with decreased risk for sudden death; however, the protective cardiovascular mechanisms of omega-3 are poorly understood. This study addresses the heart rate variability (HRV) changes in a cohort of elderly subjects randomized to receive either a daily high dose of marine-derived omega-3 fatty acids (fish oil) or a lower daily dose of a plant derived omega-3 fatty acid (alpha-linolenic acid) in soy oil. METHODS: A total of 58 elderly nursing home residents were randomized to receive 2 g/d of fish oil capsules vs 2 g/d of soy oil capsules, and were subsequently followed up every other day for a period of 6 months with 6-min measurements of HRV while resting supine. An initial control period of 2 months without supplementation was allowed to establish an HRV baseline for each participant. RESULTS: The average time- and frequency-domain parameters of HRV increased significantly during the supplementation period in both the fish oil and soy oil groups. In the regression model after adjusting for age and mean heart rate, supplementation with fish oil was associated with a significant increase in the high- and low-frequency components, and SD of normal RR intervals (SDNN), whereas only SDNN increased significantly in the soy oil group. CONCLUSIONS: Supplementation with 2 g/d of fish oil was well tolerated and was associated with a significant increase in HRV. Supplementation with 2 g/d of soy oil was associated with a lesser but significant increase in HRV. PMID- 15821182 TI - The effect of residential exercise training on baroreflex control of heart rate and sympathetic nerve activity in patients with acute myocardial infarction. AB - STUDY OBJECTIVES: Exercise training has been shown to favorably affect the prognosis after acute myocardial infarction (AMI), but the mechanisms of such favorable effects remain speculative. The aim of this study was to determine whether exercise training improves baroreflex control of heart rate and muscle sympathetic nerve activity (MSNA) in patients with AMI. DESIGN: Prospective randomized clinical study. PARTICIPANTS: Thirty patients with an uncomplicated AMI were randomized into trained or untrained groups. Arterial BP, heart rate, and MSNA were measured at rest, and during baroreceptor stimulation (phenylephrine infusion) and baroreceptor deactivation (nitroprusside infusion). These measurements were performed at baseline and after 4 weeks of exercise training. MEASUREMENTS AND RESULTS: Peak oxygen uptake increased significantly (12.3 +/- 10.7% [mean +/- SD]) with exercise training. Resting MSNA reduced from 34 +/- 12 to 27 +/- 8 bursts/min in the trained group but not in the untrained group. Arterial baroreflex sensitivity (BRS) [from 8.9 +/- 3.0 to 10.3 +/- 3.0 ms/mm Hg, p < 0.05] and MSNA response to baroreceptor stimulation (change of integrated MSNA from - 47 +/- 23 to - 70 +/- 21%, p < 0.01) improved significantly in the trained group, but not in the untrained group. Despite baroreceptor deactivation improving MSNA response in both groups, there was no significant difference between the two groups. CONCLUSIONS: Exercise training increased arterial BRS and decreased sympathetic nerve traffic after AMI, which indicate that the sympathoinhibitory effect of exercise training may, at least in part, contribute to the beneficial effect of exercise training in patients with AMI. PMID- 15821183 TI - Preinfarction angina limits myocardial infarction size in nondiabetic patients treated with primary coronary angioplasty. AB - OBJECTIVE: To evaluate myocardial necrosis extent after myocardial infarction (MI) and reperfusion with primary coronary angioplasty in nondiabetic patients and the relationship with unstable preinfarction angina (PA). DESIGN: Prospective cohort study. SETTING: Studies suggest PA limits infarct size. This effect is questioned in patients treated with primary coronary angioplasty. PATIENTS: Seventy-eight, nondiabetic, consecutive MI patients. INTERVENTIONS: Primary coronary angioplasty and scintigraphic study to assess the myocardial infarct size. MAIN OUTCOME MEASURES: Scintigraphic myocardial infarct size. RESULTS: There were 32 patients with PA (PA +) and 46 without PA (PA -) in the 24-h period prior to MI onset. There were no significant differences in the baseline characteristics between the two groups. The scintigraphy indicated myocardial infarct size significantly smaller in PA + patients: mean, 18.0% (SD, 14.7) vs 27.0% (SD, 20.1) [p = 0.033]. This occurs even though Thrombolysis in Myocardial Infarction grade 3 flow achieved in both groups was similar (84.8% vs 84.4%, p = 1.000). We found a higher percentage of ST-segment resolution (>/= 70%) in PA + patients (65.6% vs 45.7%, p = 0.082) together with a lower incidence of left ventricular systolic dysfunction (3.2% vs 18.6%, p = 0.071). CONCLUSIONS: PA exerts a beneficial effect in nondiabetic patients with ST-segment elevation acute MI even when treated with primary PCI. The infarct size is limited, and left ventricular systolic function is preserved. The effects may be related to a better preservation of tissue reperfusion in patients with PA. PMID- 15821184 TI - Does thoracic bioimpedance accurately determine cardiac output in COPD patients during maximal or intermittent exercise? AB - STUDY OBJECTIVES: The monitoring of cardiac output (CO) during exercise rehabilitation in patients with COPD, often including strenuous exercise, is advisable. Invasive methods (thermodilution, Fick method) are accurate, but for clinical routine use noninvasive CO estimation is required. We have shown that impedance cardiography (Physio Flow; Manatec Biomedical; Macheren, France) is reliable in COPD patients at rest and during a recumbent, light-intensity exercise. The aim of our study was to evaluate the validity of this noninvasive device in COPD patients during a maximal incremental exercise test (IET) and also during a strenuous intermittent work exercise test (IWET). DESIGN: Prospective comparative study of the impedance cardiograph vs the direct Fick method applied to oxygen. PATIENTS: Eight patients with moderate-to-severe COPD (59 +/- 6 years old; FEV(1), 38 +/- 15% predicted; residual volume, 194 +/- 64% predicted) [mean +/- SD]. MEASUREMENTS AND MAIN RESULTS: Forty-nine simultaneous measurements of CO by means of the direct Fick method (COfick) and CO measured by the impedance cardiograph (COpf) were obtained during the IET, and 108 measurements were made during the IWET. The correlation coefficients between the two measurements were r = 0.85 and r = 0.71 for the IET and the IWET, respectively. COpf was higher than COfick. The difference between the two methods was 3.2 +/- 2.9 L/min during the IET and 2.5 +/- 2.1 L/min during the IWET. Expressed as a percentage of the mean of the two measurements, this corresponded to 31 +/- 21% and 25 +/- 20%, respectively. CONCLUSIONS: The relatively high number of values differing by > 20% precludes the use of impedance cardiography in clinical routine in such a difficult setting (hyperinflated patients and intense exercise). PMID- 15821185 TI - Stage of lung cancer in relation to its size: part 1. Insights. PMID- 15821186 TI - Stage of lung cancer in relation to its size: part 2. Evidence. AB - OBJECTIVE: To assess the relationship between tumor size and disease stage at the time of diagnosis in non-small cell lung cancer. METHODS: From the Surveillance, Epidemiology and End Results registry, we identified all cases of primary non small cell lung cancer diagnosed prior to autopsy. Among these, we focused on 84,152 cases diagnosed in 1988 or later and documented as to tumor size and disease stage at diagnosis. The distribution of disease stage within categories of tumor size was determined. RESULTS: The smaller the tumor was, the more likely the disease was stage I. For tumors < 15 mm in diameter, the proportion of stage I was 54%, compared with 46% for 16 to 25 mm, 34% for 26 to 35 mm, 25% for 36 to 45 mm, and 15% for cases > 45 mm, with all of these proportions being very precise. CONCLUSIONS: Tumor size has substantial bearing on disease stage in non small cell lung cancer. PMID- 15821187 TI - Lung cancer detection in patients with airflow obstruction identified in a primary care outpatient practice. AB - INTRODUCTION: This prospective study describes a community-based lung cancer identification project focusing on high-risk patients who receive general care in a primary care outpatient practice. Within 1 calendar year, a simple questionnaire was completed in 1,296 patients > 50 years old to identify 430 patients at high risk of lung cancer (smoking, family history of aerodigestive tract cancer, or occupational exposures). Spirometric abnormalities were found in 126 of these patients. METHODS: Chest posteroanterior radiographs, thoracic CT scans, and sputum cytology were offered to subjects with airflow obstruction (n = 126). Eighty-eight patients underwent all tests. Thirty-eight patients refused or could not consent in a timely fashion. RESULTS: Six cancers were found in the screened group, and all were treated. Two more cancers were found in the nonscreened patients with airflow obstruction. Both were treated by surgical resection or radiation therapy. Costs per cancer found were $11,925 per patient. CONCLUSIONS: Case finding in high-risk patients in a primary care population can be accomplished at a relatively low cost. PMID- 15821188 TI - Como international conference position statement: lung cancer screening for early diagnosis 5 years after the 1998 Varese conference. AB - BACKGROUND: Lung cancer is the most common cause of cancer death in the world. Nonetheless, public policy organizations have consistently recommended against screening for lung cancer, with the result that screening is not widely practiced. The Como Conference was undertaken to consider the need for a change in the existing recommendations against screening. PURPOSES: The primary objective of the Como Conference was to consider whether there is sufficient scientific evidence to advise screening for lung cancer among asymptomatic individuals outside the context of a clinical trial. Methodological issues that are relevant to the proper interpretation of early detection trials were carefully considered. Advantages and problems associated with technological advances in CT scans and digital chest radiographs (CXRs) were fully explored. Economic issues relevant to screening were also considered. RECOMMENDATIONS: It is recommended that physicians assume responsibility for informing high-risk individuals regarding options for screening for lung cancer. Targeted high-risk individuals include middle-aged or elderly men and women who are current or former cigarette smokers of > 20 to 30 pack-years without serious medical comorbidities. It is recommended that such persons be informed that symptomatic lung cancer is usually advanced and incurable, while surgery for early lung cancer offers a far better chance of cure. They should also be informed about advances in imaging technology, as they relate to CT scans and CXRs. CONCLUSIONS: Whenever possible, high-risk individuals should be encouraged to enroll in ongoing trials. For subjects who, though eligible, do not have access to such trials, a process of shared decision-making between physicians and at-risk individuals is strongly recommended. After discussion of the existing state of knowledge, high-risk individuals should be made aware that it is reasonable for them to choose to undergo testing for lung cancer. PMID- 15821189 TI - Second primary tumors involving non-small cell lung cancer: prevalence and its influence on survival. AB - OBJECTIVES: Patients with lung cancer have a relative high risk for second primary cancers. We studied the prevalence of second primary tumors in patients with a diagnosis of non-small cell lung cancer (NSCLC) in their history or at follow-up. Furthermore, we studied survival in subgroups of those patients. METHODS AND PATIENTS: Retrospectively, 860 patients with NSCLC that had been diagnosed in the period from January 1, 1990, to December 31, 1999, were evaluated for second primary cancers either in their history or in the follow-up period. The patients were divided into the following four groups: group I, patients with another primary tumor detected in the follow-up period (n = 44); group II, patients with another primary tumor in their history (n = 148); group III, patients with no other primary tumor found in their history or at follow-up (n = 634); and group IV, patients with more than one other primary tumor in found in their history or at follow-up (n = 34). RESULTS: The most frequently diagnosed double tumors were located in the lungs, the head and neck region, and the urinary tract. The interval between another malignancy as the first tumor (group II, 83 months) and NSCLC as the second malignancy was significantly longer than vice versa (group I, 14.5 months; p < 0.05). In > 80% of patients, the second primary tumors were diagnosed within 1 year after NSCLC was diagnosed. The 5-year survival rate is significantly better for patients with more than two primary malignancies compared to patients without two primary malignancies and patients with one other tumor in their history (p = 0.004 and 0.012, respectively). The 5 year survival rate in patients with a second tumor in the follow-up period was better than in patients without any other second tumor (p = 0.029). As the TNM stage and therapy were comparable in all subgroups, it could not be used as explanation for the difference in survival rates. CONCLUSION: In 25% of patients, additional tumors that were NSCLC were diagnosed either in their history or in the follow-up period. The majority of second tumors following NSCLC are diagnosed within 1 year. Nevertheless, patients with a second tumor tend to have an overall better survival rate than patients without second primaries, suggesting different growth habits. PMID- 15821190 TI - Cardiopulmonary exercise tests and lung cancer surgical outcome. AB - STUDY OBJECTIVES: Surgical resection remains the treatment of choice for anatomically resectable non-small cell lung cancer. However, the presence of associated comorbid conditions increases the risk of death and surgical complications. Several studies have evaluated the usefulness of preoperative exercise testing for predicting postoperative morbidity and mortality. The aim of this study was to establish whether exercise testing could predict poor surgical outcome in lung cancer surgery and whether the absolute value or percentage of predicted value is the better predictor of the surgical outcome. DESIGN: The study was designed as a prospective study. PATIENTS AND SETTING: One hundred thirty patients with potentially operable lung cancer at Papworth Hospital over 2 years were recruited; of these, 101 underwent curative surgery. INTERVENTIONS: Spirometry and cardiopulmonary exercise tests were performed for every patient (n = 99), except for two patients with back problems. We also recorded the outcome of surgery, in particular, complications and mortality. MEASUREMENTS AND RESULTS: Mean maximum oxygen transport at peak exercise (Vo(2)peak) was 18.3 mL/kg/min (SD, 4.7 mL/kg/min), and mean percentage of predicted Vo(2)peak value was 84.4% (SD, 30%). Poor surgical outcome was significantly related to Vo(2)peak percentage of predicted (p < 0.01) but not to the actual oxygen uptake value. CONCLUSIONS: The use of the percentage of predicted Vo(2)peak value would be a better indicator of surgical outcome, since it predicts the surgical outcome better, and corrects for normal physiologic ranges. The threshold of Vo(2)peak for surgical intervention could be set between 50% and 60% of predicted without excess surgical mortality. PMID- 15821192 TI - Causes and presenting features of pulmonary infarctions in 43 cases identified by surgical lung biopsy. AB - BACKGROUND: Although pulmonary infarction is usually associated with pulmonary thromboembolism, it can occur with other disorders such as vasculitis, angioinvasive infections, sickle-cell disease, tumor embolism, and pulmonary torsion. STUDY OBJECTIVE: To identify causes and presenting features of pulmonary infarctions diagnosed by surgical biopsy in a consecutive series of patients encountered at a single institution. DESIGN: Retrospective review. SETTING: Tertiary care, referral medical center. PATIENTS: Forty-three patients with pulmonary infarction identified on surgical lung biopsy over a period of 7 years, January 1996 through December 2002. RESULTS: The median age of these 43 patients was 55 years (range, 22 to 85 years); 17 patients (40%) were women, and 26 patients (60%) were men. Thirty-five patients (81%) had a smoking history. Twenty eight patients (65%) presented with solitary or multiple lung nodules/masses of undetermined etiology. The underlying cause was identifiable in 31 cases (72%) based on a review of clinical, laboratory, radiologic, and histopathologic data. The two most common causes were pulmonary thromboembolism (18 cases, 42%) and pulmonary infections (5 cases, 12%). Thromboembolic pulmonary infarctions typically presented as solitary or multiple nodules located in the subpleural regions. Other causes included diffuse alveolar damage in two cases (5%), pulmonary torsion in two cases (5%), and one case each of lung cancer, amyloidosis, embolotherapy, and catheter embolism. In 12 cases (28%), the underlying cause was not directly identifiable but was probably due to previous pulmonary thromboembolism. CONCLUSION: We conclude that although pulmonary thromboembolism is the most common cause of pulmonary infarction identified by surgical lung biopsy, a variety of other causes are clinically encountered, including infections, inflammatory or infiltrative lung diseases, pulmonary torsion, malignancy, and nonthrombotic embolism. Pulmonary infarction should be considered in the differential diagnosis of peripheral lung nodules or masses. PMID- 15821191 TI - Lung volume reduction surgery vs medical treatment: for patients with advanced emphysema. AB - OBJECTIVE: To contribute to the knowledge on the therapeutic value of lung volume reduction surgery (LVRS). DESIGN: Two similar, independently conceived and conducted, multicenter, randomized clinical trials. SETTING: The Canadian Lung Volume Reduction (CLVR) study and the Overholt-Blue Cross Emphysema Surgery Trial (OBEST). METHODS: Using a fixed-effects meta-analysis, the 6-month results produced by the addition of LVRS to optimal medical therapy were compared to those obtained from optimal medical therapy alone. Patients were required to have severe emphysema, marked airflow limitation (ie, FEV(1), 15 to 40% predicted), hyperinflation (total lung capacity [TLC], > 120% predicted), CO(2), < 55 mm Hg, and measurable dyspnea (chronic respiratory disease questionnaire [CRDQ] scores /= 1 for the OBEST). Optimal medical therapy included pulmonary rehabilitation in both arms of both studies. RESULTS: The CLVR study randomized 58 patients and the OBEST randomized 35 patients for a total of 93 patients. Of these, 54 patients were randomized to undergo surgery, and 39 patients were randomized to receive medical treatment. The 6-month mortality rate (including operative mortality) in the surgical and medical cohorts was similar (5.6% vs 5.1%, respectively). A comparison of the medical and surgical arms of the combined CLVR study/OBEST population showed that LVRS was associated with a higher FEV(1) (167 mL or 24% predicted; 95% confidence interval [CI], 29 to 304; p = 0.017), lower residual volume (-1,342 mL or 24.5% predicted; 95% CI, -1,844 to -840; p < 0.001), lower TLC (-1,044 mL or 13% predicted; 95% CI, -1483 to -605; p < 0.001), and higher 6 min walk distance (148.8 feet; 95% CI, 24.3 to 273.2; p = 0.019). Each domain of the CRDQ showed statistically significant improvement in the surgical arm of the study, but not in the medical arm. The summary physical component scale of the Medical Outcomes Study 36-item short form (SF-36) was also more favorable in the LVRS cohort (6.9; 95% CI, 2.86 to 10.90; p < 0.001). The summary mental component scale of the SF-36 did not show a statistically significant difference between the two groups. CONCLUSION: Six months after randomization, LVRS produced better palliation than optimal medical therapy in patients with advanced emphysema. PMID- 15821193 TI - Plasma L-arginine and metabolites of nitric oxide synthase in patients with left to-right shunt after intracardiac repair. AB - STUDY OBJECTIVE: Human plasma L-arginine serves as a substrate pool for endothelial-derived nitric oxide (NO) synthase. In this pilot study, we tested the hypothesis that plasma L-arginine and other metabolites of the L-arginine NO pathway could correlate with postoperative pulmonary hypertension after cardiopulmonary bypass (CPB). DESIGN: Forty-two patients (median age, 0.5 years; range, 0.1 to 28 years) with atrial septal defect (n = 15), ventricular septal defect (n = 18), atrioventricular canal (n = 8), and aortopulmonary window (n = 1) were enrolled. The influence of patient age, preoperative pulmonary hypertension, duration of CPB, plasma L-arginine, guanosine 3', 5'-cyclic monophosphate (cGMP), and nitrate on postoperative pulmonary hypertension during the first 24 h after CPB was studied by logistic regression. RESULTS: Nineteen of 42 patients were found to have preoperative pulmonary hypertension. Thirteen of 42 patients showed persistent pulmonary hypertension after intracardiac repair with a mean pulmonary artery pressure (PAP) of 38 mm Hg (range, 23 to 55 mm Hg) at 24 h after CPB. L-arginine concentrations in plasma were significantly lower 24 h after CPB than before: 52 mumol/L (range, 18 to 95 mumol/L) vs 79 mumol/L (range, 31 to 157 mumol/L). Plasma cGMP levels were higher and plasma nitrate levels were lower immediately after weaning from CPB (p < 0.0033). On logistic regression analysis, only patient age (p = 0.02) and preoperative PAP (p = 0.01) were related to postoperative pulmonary hypertension. CONCLUSION: Low plasma L arginine does not relate to persistent pulmonary hypertension in patients with left-to-right shunt after CPB and intracardiac repair. PMID- 15821194 TI - Pneumoproteins as a lung-specific biomarker of alveolar permeability in conventional on-pump coronary artery bypass graft surgery vs mini-extracorporeal circuit: a pilot study. AB - BACKGROUND: Despite improvements of the heart-lung machine (HLM), oxidative stress and subsequent damage to the alveolar capillary membrane still occur after conventional on-pump coronary artery bypass graft (CCABG) surgery. In an attempt to further improve the conventional HLM, a mini-extracorporeal circuit (MECC) was introduced. This new concept is based on minimal volume shifts. The extent of alveolar injury that is associated with this new technique is unknown. The lung specific biomarkers Clara-cell 16 (CC16) and KL-6 are applied in this study to quantify alveolar dysfunction in both techniques. METHODS: In a prospective observational setting, the concentrations of CC16 and KL-6 were measured during and after 10 consecutive CCABG operations and 10 consecutive coronary artery bypass graft (CABG) operations using MECC (MCABGs). These pneumoproteins were measured after the induction of anesthesia, before clamping of the ascending aorta, after unclamping of the aorta, on arrival to the ICU, and on the following days until discharge. Quantification of the differences of KL-6 and CC16 leakage through the alveolar membranes between the two techniques was realized by calculation of the Student t test. Perioperative and postoperative shunt fractions and clinical observations were monitored simultaneously. The potential value of pneumoproteins as biomarkers for quantification of alveolar permeability during CABG surgery was tested. RESULTS: Significantly reduced concentrations of CC16 were found early after MCABG as compared to CCABG surgery (p = 0.033). KL-6 showed no consistent pattern during both treatment modalities. Early after CCABG surgery, shunt fractions tended to show reduced oxygen transport over the alveolar membrane as compared to MCABG surgery. CONCLUSION: CC16 appears to be a useful biomarker for alveolar permeability during CABG surgery. Injury of the alveolar capillary membrane appears significantly reduced during MCABG surgery. Consistently early postoperative alveolar shunt fractions showed an increased value in CCABG compared to MCABG surgery in the early postoperative phase. Further randomized studies need to confirm the value of CC16 as marker in monitoring alveolar capillary damage during coronary bypass grafting. PMID- 15821195 TI - Mortality in individuals with severe deficiency of alpha1-antitrypsin: findings from the National Heart, Lung, and Blood Institute Registry. AB - STUDY OBJECTIVE: To clarify the mortality rate and causes of death of individuals with alpha(1)-antitrypsin (AAT) deficiency, the Death Review Committee (DRC) of the National Heart, Lung, and Blood Institute Registry of Individuals with Severe AAT Deficiency reviewed all available medical records regarding the deaths of study subjects during Registry follow-up (up to 7.2 years). METHODS: Individual determinations by each member of the three-person DRC led to consensus judgments regarding the underlying cause and the immediate and contributing causes of death. RESULTS: Of the 1,129 Registry subjects, 204 died (18.1%) [approximately 3%/yr]. Record availability permitted detailed review in 120 decedents, and death certificates were available in 56 of the remaining 84 subjects (67%). Emphysema and cirrhosis were the most common underlying causes of death (72% and 10%, respectively), with malignancy and diverticulitis accounting for 3% of deaths each. To assess attributable mortality, standardized mortality ratio analysis was performed and indicated that excess mortality was ascribable entirely to lung and liver disease. CONCLUSIONS: We conclude that severe AAT deficiency poses a significant threat to health, that severe airflow obstruction is a major determinant of mortality, and that liver and lung disease account for the excess mortality in affected individuals. These findings support current efforts to enhance diagnostic recognition and treatment of AAT-deficient individuals. PMID- 15821196 TI - Surprisingly high prevalence of anxiety and depression in chronic breathing disorders. AB - STUDY OBJECTIVES: The objectives of this study were to assess the prevalence, screening, and recognition of depression and anxiety in persons with chronic breathing disorders, including COPD. DESIGN: Cross-sectional study. SETTING: The Michael E. DeBakey Veterans Affairs Medical Center (MEDVAMC). PARTICIPANTS: A large sample of 1,334 persons with chronic breathing disorder diagnoses who received care at the MEDVAMC. MEASUREMENTS: The prevalence of anxiety and depression was measured in a large sample of persons with a chronic breathing disorder diagnosis who received care at the MEDVAMC, using the Primary Care Evaluation of Mental Disorders (PRIME-MD) screening questions. The positive predictive value of the PRIME-MD questions was then determined. The prevalence of anxiety and depressive diagnoses in patients determined to have COPD was then measured, using the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (SCID). RESULTS: Of patients screened with the PRIME-MD, 80% screened positive for depression, anxiety, or both. The predictive value of a positive phone screen for either depression or anxiety was estimated to be 80%. In the subsample of patients who had COPD and received a diagnosis using the SCID, 65% received an anxiety and/or depressive disorder diagnosis. Of those patients, only 31% were receiving treatment for depression and/or anxiety. CONCLUSIONS: It is troubling that a mere 31% of COPD patients with depression or anxiety are being treated, particularly given their high prevalence in this population. Practical screening instruments may help increase the recognition of anxiety and depression in medical patients, as suggested by the excellent positive predictive value of the PRIME-MD in our study. PMID- 15821197 TI - Treatment with the immunomodulator AM3 improves the health-related quality of life of patients with COPD. AB - BACKGROUND: COPD has a severe impact on patient quality of life. AM3 is an orally effective immunomodulator that can normalize the defective antimicrobial functions of the immune system effector cells of COPD patients. OBJECTIVES: We analyzed the effect of AM3 on exacerbation frequency and health-related quality of life (HRQL) of COPD patients with moderate disease. DESIGN: A randomized, double-blind, placebo-controlled trial. SETTING: Outpatient departments of 21 hospitals. METHODS: A total of 253 COPD patients with a mean age of 67.7 years (SD, 8.1 years) and mean FEV(1) percentage of predicted of 49.6% (SD, 10.2%) were evaluated. Patients received (orally) either 3 g/d AM3 or a matched placebo for 180 consecutive days. Patient quality of life was measured using the St. George's Respiratory Questionnaire (SGRQ). RESULTS: There were no differences in the exacerbation frequency of the two groups (0.82 episodes per patient in the AM3 arm vs 0.84 in the placebo arm), and 55.3% of patients were exacerbation free in the AM3 arm compared to 48.8% in the placebo arm (p = 0.11). At the end of treatment, quality of life was significantly better in the AM3 arm than in the placebo arm (SGRQ total score, 32.9; SD, 16.4, compared to 37.5; SD, 17.5 [p < 0.05]: activity score, 47.5; SD, 22.4, compared to 54.6; SD, 20.5 [p < 0.05]). The improvements in total SGRQ scores were 8.9 U (SD, 13.4 U) in the AM3 arm and 5.6 U (SD, 15.9 U) in the placebo arm (p = 0.076). Improvements on the symptoms subscale were 15.9 U (SD, 20.7 U) for the AM3 arm and 10.2 U (SD, 21.3 U) for the placebo arm (p < 0.05). Both AM3 and the placebo were clinically, biochemically, and hematologically well tolerated. CONCLUSIONS: AM3 is a safe, easily tolerated, effective treatment that improves the quality of life of COPD patients as measured by SGRQ scores. This effect was observed with no significant reduction in the frequency of exacerbations. PMID- 15821198 TI - Relationships among bacteria, upper airway, lower airway, and systemic inflammation in COPD. AB - STUDY OBJECTIVE: The upper and lower airways are continuous. While upper airway symptoms are common in COPD patients, with accumulating evidence to suggest increased nasal inflammation, the relationships among upper airway, lower airway, and systemic inflammatory indexes have not been studied. We aimed to confirm that there is heightened nasal inflammation in COPD patients, to test the hypothesis that the degree of upper airway inflammation relates to the degree of lower airway inflammation, and to investigate the underlying associations with bacterial carriage and the systemic inflammatory response. DESIGN: Prospective cohort study. SETTING: Outpatient Department, London Chest Hospital, London, UK. PARTICIPANTS: Forty-seven patients with COPD and 12 control subjects of similar age, sex, and smoking status. MEASUREMENTS: Serum, nasal wash fluid, and sputum samples were obtained from 47 stable patients with COPD for the analysis of inflammatory indexes and bacterial colonization. Nasal wash fluid specimens were obtained from 12 control subjects. RESULTS: COPD patients had an increased nasal interleukin (IL)-8 concentration compared to control subjects (difference, 97.2 pg/mL; p = 0.009). The nasal IL-8 concentration in COPD patients correlated with that in sputum (r = 0.30; p = 0.039). In both the upper and lower airways of patients with COPD, the IL-8 concentration was associated with indexes of bacterial colonization. Patients colonized with a sputum potentially pathogenic microorganism had a higher total nasal bacterial load (difference, 1.5 log cfu/mL; p = 0.016). We did not find significant relationships between the degree of upper or lower airway inflammation, or bacterial carriage, and the systemic inflammatory response. CONCLUSIONS: COPD is associated with an increased nasal concentration of the neutrophil chemoattractant protein IL-8, the degree of which reflects that present in the lower airway. A relationship between lower airway bacterial colonization, postnasal drip, and higher nasal bacterial load may suggest a mechanism underlying this finding. This study is the first to report a correlation between the degree of upper and lower airway inflammation in COPD. PMID- 15821199 TI - Prevalence of gastroesophageal reflux in difficult asthma: relationship to asthma outcome. AB - STUDY OBJECTIVES: To determine the prevalence of gastroesophageal reflux disease (GERD)-both symptoms and objective evidence-using 24-h dual-probe pH monitoring in difficult asthma, and the relationship between the presence and treatment of GERD to clinical outcome. DESIGN AND SETTING: As part of a systematic evaluation protocol, 68 subjects with difficult-to-control asthma attending a difficult asthma clinic were referred for dual-probe ambulatory pH esophageal monitoring. RESULTS: Esophageal probe data were available in 52 patients (76%) with difficult asthma. The prevalence of GERD/GERD-associated asthma symptoms was 75% (39 of 52 patients; 95% confidence interval [CI], 63 to 84.7%). The prevalence of GERD as evidenced by an abnormal pH profile at the distal esophageal probe was 55% (29 of 52 patients; 95% CI, 40 to 69%). The prevalence of GERD at the proximal probe was 34.6% (18 of 52 patients; 95% CI, 23.6 to 51%). The prevalence of GERD was similar in asthmatic subjects who responded to intervention and those who remained difficult to control (therapy resistant). Asymptomatic GERD was present in 9.6% (5 of 52 patients); 16% of cough episodes correlated with acid reflux. CONCLUSIONS: In difficult-to-control asthma, GERD is common, but identification and treatment of GERD do not appear to relate to improvement in asthma control in this population. PMID- 15821200 TI - Maternal and grandmaternal smoking patterns are associated with early childhood asthma. AB - OBJECTIVE: To investigate the associations of maternal and grandmaternal smoking before, during, and after pregnancy with childhood asthma. DESIGN, SETTING, AND PARTICIPANTS: We conducted a case-control study nested within the Children's Health Study in southern California. The case patients consisted of 338 children with asthma that had been diagnosed in the first 5 years of life, and 570 control subjects were countermatched on in utero exposure to maternal smoking within grade, sex, and community of residence. MEASUREMENTS: Detailed maternal and household smoking histories and other asthma risk factor information was obtained by telephone interview. RESULTS: The participation rates were 72.3% and 82.5%, respectively, for control subjects and case patients. In utero exposure to maternal smoking was associated with increased risk for asthma diagnosed in the first 5 years of life (odds ratio [OR], 1.5; 95% confidence interval [CI], 1.0 to 2.3), and for persistent asthma (OR, 1.5; 95% CI, 1.0 to 2.3). The associations did not differ in children with early transient asthma compared to those with early persistent asthma. Relative to never-smokers, children whose mothers smoked throughout the pregnancy had an elevated risk of asthma in the first 5 years of life (OR, 1.6; 95% CI, 1.0 to 2.6). Children of mothers who quit smoking prior to the pregnancy showed no increased risk (OR, 0.9; 95% CI, 0.5 to 1.5). We were unable to assess the association of smoking cessation during pregnancy because very few mothers were reported to have done so (15%). Asthma risk did not increase in a monotonic pattern with smoking intensity during pregnancy. Postnatal secondhand smoke exposure was not independently associated with asthma. Grandmaternal smoking during the mother's fetal period was associated with increased asthma risk in her grandchildren (OR, 2.1; 95% CI, 1.4 to 3.2). CONCLUSIONS: Maternal and grandmaternal smoking during pregnancy may increase the risk of childhood asthma. PMID- 15821201 TI - Regular use of corticosteroids and low use of short-acting beta2-agonists can reduce asthma hospitalization. AB - OBJECTIVES: Inhaled corticosteroids (ICS) and inhaled short-acting beta(2) agonists (ISABA) are the most commonly used medications for management of asthma. Increased asthma morbidity and mortality have been reported with excess use of ISABA in several studies. In these studies, authors have used different indicators to control for the potential confounding by asthma severity. The objective of this study was to determine the effect of ICS use on the association between use of ISABA and first hospitalization for asthma after controlling for several indicators of asthma severity. DESIGN: An inceptional cohort study using Saskatchewan Health databases. SETTING: The Province of Saskatchewan, Canada. PARTICIPANTS: A total of 29,957 persons aged 5 to 54 years who had at least five asthma-related visits to physicians between 1991 and 2000. RESULTS: Among the subjects with increased asthma severity, indicated by one or more average number of asthma-related physician visits per 3 months during the follow-up, high use of ISABA was a risk factor for hospitalization when no ICS were used (rate ratio [RR], 2.16; 95% confidence interval [CI], 1.51 to 2.95). There was a beneficial effect of ISABA when there was low use of ICS (RR, 0.65; 95% CI, 0.42 to 0.93) or high use of ICS (RR, 0.23; 95% CI, 0.12 to 0.41). Among the subjects with less severe asthma, indicated by less than one asthma-related physician visits per 3 months, on average, during the follow-up, the risk of hospitalization was even greater for high use of ISABA when no ICS were used (RR, 10.06; 95% CI, 6.99 to 14.47). This was reduced but not abolished when there was low use of ICS (RR, 3.24; 95% CI, 2.29 to 4.59) and negated altogether by high use of ICS (RR, 1.10; 95% CI, 0.39 to 3.12). CONCLUSION: Among both severe and less severe asthma groups, high use of ISABA was associated with an increased risk of asthma hospitalization in the absence of any use of ICS, which was progressively reduced with low and high use of ICS. This finding was independent of asthma severity and could result from lack of control through over reliance on ISABA in asthma management. PMID- 15821202 TI - Sensitization to Aspergillus antigens and occurrence of allergic bronchopulmonary aspergillosis in patients with asthma. AB - BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA), which is predominantly a disease of asthmatic subjects, is caused by hypersensitivity to Aspergillus antigens. Screening for Aspergillus sensitization in asthmatic subjects could identify those who are at risk for ABPA. Few studies have shown that fungal sensitization could be an important risk factor for asthma severity. We sought to determine the frequency of sensitization to Aspergillus antigens in asthmatic subjects and its effect on disease severity. We also determined the occurrence of ABPA in these subjects. DESIGN: Prospective study of consecutive patients with asthma. SETTING: Tertiary university referral hospital, outpatient department. PATIENTS AND METHODS: One hundred five asthmatic subjects and 26 volunteers underwent skin testing with aeroallergens, including Aspergillus, serum precipitins against Aspergillus antigens, and specific IgG against Aspergillus fumigatus, total serum IgE levels, and routine blood and radiologic investigations. ABPA was diagnosed when all eight major criteria were fulfilled. RESULTS: Thirty patients (28.5%) had a positive skin reactivity to Aspergillus antigens. Eleven patients (10.4%) had positive specific reactions to IgG, and 8 patients (7.6%) demonstrated positive reactions to serum precipitins. Eight of these 30 patients (26.6%) received diagnoses of ABPA, which was 7.6% of the total. None of the control subjects were sensitized to Aspergillus antigens. The patients were classified into the following four groups: negative skin test results; positive reactions to aeroallergens other than Aspergillus; positive reactions to aeroallergens including Aspergillus antigens; and patients with ABPA. Based on clinical and serologic parameters, patients with Aspergillus sensitive asthma and ABPA had a significantly more severe form of the disease. CONCLUSIONS: Sensitization to the mold Aspergillus increases the severity of asthma. ABPA should be excluded in all patients with Aspergillus-sensitive asthma. PMID- 15821203 TI - Factors influencing in-hospital mortality in community-acquired pneumonia: a prospective study of patients not initially admitted to the ICU. AB - PURPOSES: To determine the factors that predict in-hospital mortality among patients who require hospitalization for the treatment of community-acquired pneumonia (CAP). STUDY DESIGN: Prospective observational study of all patients who were admitted to six hospitals in Edmonton, AL, Canada, with a diagnosis of CAP from November 15, 2000, to November 14, 2002. Pneumonia was defined as two or more respiratory symptoms and signs and an opacity on a chest radiograph as interpreted by the attending physician. RESULTS: A total of 3,043 patients were enrolled in the study, 246 of whom died (8.1%). On multivariate analysis, increasing pneumonia severity score, increasing age, site of care, consultation with a respirologist or infectious diseases physician, and functional status at the time of admission were all independently predictive of mortality. Increasing pneumonia severity risk score, increasing age, site of hospitalization, functional status, and consultation with an infectious diseases physician or a respirologist were independently associated with both early (< 5 days) and late (>/= 5 days) mortality. In contrast, partial or complete use of the pneumonia pathway was associated with decreased early mortality, but had no effect on late mortality. A low lymphocyte count and a high serum potassium level were associated with early but not with late mortality. The type of antibiotic therapy had an effect on late but not on early mortality. CONCLUSIONS: Functional status at the time of hospital admission is a powerful predictor of mortality and should be incorporated into any scores or models that are used to predict mortality. While there are some common predictors of early and late in-hospital mortality, early mortality is not affected by the timing or type of antibiotic therapy, whereas late mortality is influenced by the type of antibiotic therapy. Hyperkalemia and lymphopenia are associated with early mortality. PMID- 15821204 TI - Intrathoracic lymphadenopathy in hospitalized patients with pneumococcal pneumonia. AB - PURPOSES: Pneumococcal pneumonia is the most common etiology for community acquired pneumonia. The prevalence of lymphadenopathy seen on CT scans in these patients is uncertain. The purpose of this series was to assess the prevalence of intrathoracic lymphadenopathy in hospitalized patients with pneumococcal pneumonia. MATERIALS AND METHODS: We retrospectively identified 35 hospitalized patients with pneumococcal pneumonia who had been evaluated with CT scanning between January 1998 and April 2002. There were 18 men and 17 women with a mean age of 56 years. The study inclusion criteria were as follows: blood culture positive for Streptococcus pneumoniae, clinical diagnosis of pneumonia, and chest CT scan. The study exclusion criteria were known causes of lymphadenopathy. Charts were reviewed for HIV status, smoking history, and comorbidities. CT scans were reviewed for the presence and degree of lymphadenopathy, and the characteristics of pneumonia. The number of enlarged lymph nodes was graded as few (one to two), moderate (three to five), or many (six or more). Pneumonias were described by location, the number of involved lobes, the presence of cavitation, and the presence of pleural effusion. Patients with different comorbidities and different imaging characteristics of pneumonia were compared. RESULTS: Among the 35 patients with pneumococcal pneumonia, intrathoracic lymphadenopathy was present on CT scans in 54% of patients (19 of 35 patients). The lymphadenopathy was ipsilateral to the pneumonia in 100% of patients (19 of 19 patients). One patient also had contralateral lymphadenopathy. The lymphadenopathy was graded as few in 37% of patients (7 of 19 patients), moderate in 37% of patients (7 of 19 patients), and many in 26% of patients (5 of 19 patients). The pneumonia was right-sided in 26% of patients (9 of 35 patients), left-sided in 17% of patients (6 of 35 patients), and bilateral in 57% of patients (20 of 35 patients). Comorbidities included the following: HIV infection (n = 15); smoking (n = 21); emphysema (n = 5); hepatitis C (n = 5); and diabetes (n = 3). The characteristics of pneumonia included the following: multilobar (n = 25); pleural effusion (n = 25); and cavitation (n = 5). The prevalence of lymphadenopathy according to comorbidities and characteristics of pneumonia ranged from 40 to 100%. None of the differences in prevalence of lymphadenopathy among the subgroups was statistically significant. CONCLUSION: Lymphadenopathy is a common CT scan feature of pneumococcal pneumonia in hospitalized patients, in a variety of settings. Therefore, when a patient with pneumococcal pneumonia has lymphadenopathy seen on a CT scan, other etiologies for the lymphadenopathy need not be suspected. PMID- 15821205 TI - Reappraisal of clindamycin IV monotherapy for treatment of mild-to-moderate aspiration pneumonia in elderly patients. AB - BACKGROUND: As the number of elderly people has increased in Japan, the occurrence of aspiration pneumonia has also increased. Guidelines for the treatment of pneumonia have been proposed, in which the use of antibiotics, such as beta-lactam plus beta-lactamase inhibitor, clindamycin, and carbapenem, has been recommended as effective against anaerobic bacteria in the treatment of aspiration pneumonia. However, to our knowledge, a prospective comparison of these antibiotics regarding their clinical efficacy in aspiration pneumonia has not been performed. STUDY OBJECTIVES: We compared the effects of IV administration of a half dose of ampicillin/sulbactam (SBT/ABPC), normal dose of SBT/ABPC, IV clindamycin, and IV panipenem/betamiprom (PAPM/BP) for treatment of mild-to-moderate aspiration pneumonia in elderly patients. DESIGN: Randomized prospective study. PATIENTS: One hundred adult patients with compatible signs and symptoms of aspiration pneumonia. ASSESSMENTS: Patients were assessed before, during, and after treatment regarding symptoms, as well as results of laboratory values, chest radiograph examinations, and sputum bacterial cultures. RESULTS: We found few differences between the groups regarding cure rate, duration of IV medication, and occurrence of adverse effects with the tested therapies. However, clindamycin therapy was less expensive and was associated with a lower rate of posttreatment occurrence of methicillin-resistant Staphylococcus aureus. CONCLUSIONS: Clindamycin therapy for mild-to-moderate aspiration pneumonia is clinically effective, and provides economic advantages as compared to SBT/ABPC or PAPM/BP therapy. PMID- 15821206 TI - Identification of mycobacterium species in contaminated cultures by polymerase chain reaction. AB - STUDY OBJECTIVES: The detection of Mycobacterium sp on a culture remains the "gold standard" technique for the diagnosis of mycobacterial infections. A small percentage of these cultures, however, may be contaminated by other nonfastidious microorganisms, making accurate diagnosis difficult. We evaluated the use of a polymerase chain reaction (PCR) protocol that was specific for the genus Mycobacterium, and specifically for Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium intracellulare, for the identification of Mycobacterium sp growing on contaminated cultures. DESIGN: This prospective study was designed to identify Mycobacterium sp growing on mycobacterial cultures contaminated with other microorganisms. Samples and patients: Twenty-six samples, taken from 23 patients with probable mycobacterial disease, that resulted in Mycobacterium growth but were contaminated during their processing were evaluated in this study. Clinical data and the clinical status of each patient were used to ascertain the final diagnosis. RESULTS: All samples studied here exhibited Mycobacterium growth on solid media but were contaminated by nonfastidious bacteria, compromising the biochemical identification of the Mycobacterium sp. PCR correctly identified the genus Mycobacterium in all samples. M tuberculosis was identified in 14 samples, and M avium in 10 samples. No amplification of M intracellulare was obtained, and in two samples there was amplification only for the genus Mycobacterium. In the cultures of those patients in whom a mycobacterial infection was evident, PCR identified M avium and M tuberculosis in samples from 6 and 12 patients, respectively. However, PCR identified M avium (two patients) and M tuberculosis (two patients) in the cultures of four patients for whom a mycobacterial disease could not be confirmed by our case definition. Finally, in two samples from one patient only the genus Mycobacterium was amplified by PCR. CONCLUSION: PCR, with its advantages of greater speed and effectiveness than conventional detection methods, was successfully used to identify the Mycobacterium sp growing on contaminated cultures. PMID- 15821207 TI - Tuberculosis during fundamental societal changes in Estonia with special reference to extrapulmonary manifestations. AB - STUDY OBJECTIVES: To characterize the incidence of extrapulmonary tuberculosis (EPTB) in Estonia, by site and age, in conditions of marked pulmonary tuberculosis (TB) increase, ie, during the period of fundamental societal changes, but where TB incidence has not been influenced by HIV infection. DESIGN AND SETTING: A retrospective study. PATIENTS: All new cases of EPTB (n = 622) detected in Estonia from 1991 to 2000. RESULTS: The incidence of pulmonary TB in Estonia increased from 21.5 in 1991 to 44.6/100,000 in 2000. In contrast, the incidence of EPTB did not increase at the same pace, changing only from 3.6 in 1991 to 4.7/100,000 in 2000. The increase in the incidence of EPTB was significantly lower than that of pulmonary TB (p < 0.05), and the relative frequency of EPTB among overall TB steadily decreased from 17.0% in 1991 to 10.1% in 2000. The incidence of EPTB forms with a long latency period (eg, osteoarticular and urogenital TB) remained unchanged, while that of EPTB forms with a short latency period (eg, intrathoracic lymph node TB) increased (p < 0.05). The number of cases of urogenital, pleural, and osteoarticular TB increased with age; the number of cases of intrathoracic lymph node TB decreased with age. The bacteriologic confirmation rate was lower in EPTB than in pulmonary TB and varied according to site. CONCLUSION: There was no increase in the incidence of EPTB during 10 years despite the dramatic increase in overall TB. The proportion of EPTB among all TB tended to decrease with increasing age, and different EPTB sites prevailed in different age groups. PMID- 15821208 TI - Adverse events and treatment completion for latent tuberculosis in jail inmates and homeless persons. AB - BACKGROUND: Recently, a short-course treatment using 60 daily doses of rifampin and pyrazinamide was recommended for latent tuberculosis (TB) infection (LTBI). STUDY OBJECTIVES: To determine the acceptability, tolerability, and completion of treatment. DESIGN: Observational cohort study. SETTING: Five county jails and TB outreach clinics for homeless populations in three cities. PATIENTS: Study staff enrolled 1,211 patients (844 inmates and 367 homeless persons). INTERVENTIONS: Sites used 60 daily doses of rifampin and pyrazinamide, an approved treatment regimen for LTBI. MEASUREMENTS: Types and frequency of drug-related adverse events and outcomes of treatment. RESULTS: Prior to treatment, 25 of 1,178 patients (2.1%) had a serum aminotransferase measurement at least 2.5 times the upper limit of normal. Patients who reported excess alcohol use in the past 12 months were more likely than other patients to have an elevated pretreatment serum aminotransferase level (odds ratio, 2.1; 95% confidence interval, 1.1 to 6.1; p = 0.03). Treatment was stopped in 66 of 162 patients (13.4%) who had a drug-related adverse event. Among 715 patients who had serum aminotransferase measured during treatment, 43 patients (6.0%) had an elevation > 5 times the upper limits of normal, including one patient who died of liver failure attributed to treatment. In multivariate analyses, increasing age, an abnormal baseline aspartate aminotransferase level, and unemployment within the past 24 months were independent risk factors for hepatotoxicity. Completion rates were similar in jail inmates (47.5%) and homeless persons (43.6%). CONCLUSIONS: This study detected the first treatment-associated fatality with the rifampin and pyrazinamide regimen, prompting surveillance that detected unacceptable levels of hepatotoxicity and retraction of recommendations for its routine use. Completion rates for LTBI treatment using a short-course regimen exceeds historical rates using isoniazid. Efforts to identify an effective short-course treatment for LTBI should be given a high priority. PMID- 15821209 TI - Inactive hepatitis B surface antigen carrier state and hepatotoxicity during antituberculosis chemotherapy. AB - STUDY OBJECTIVES: To determine whether inactive hepatitis B surface antigen (HBsAg) carriers are at a higher risk of drug-induced hepatotoxicity than control subjects during antituberculosis treatment with standard short-course regimens of isoniazid, rifampin, ethambutol, and/or pyrazinamide. DESIGN: Retrospective case control study. SETTING: Tertiary university medical center. PATIENTS: One hundred ten inactive HBsAg carriers with newly diagnosed active tuberculosis who had been treated with isoniazid, rifampin, ethambutol, and/or pyrazinamide were included in the study population. Inactive HBsAg carriers were defined as follows: (1) positive for HbsAg; (2) negative for hepatitis B e antigen (HBeAg), positive for antibody to HBeAg; (3) < 10(5) copies per mL of serum hepatitis B virus DNA; and (4) normal pretreatment aspartate aminotransferase (AST)/alanine aminotransferase (ALT) levels. Ninety-seven HBsAg-negative patients who received standard antituberculosis medication were selected as control subjects. RESULTS: The baseline characteristics of the 110 inactive HBsAg carriers were similar to those of the 97 noncarriers. A total of 85% of persons in both groups had received an initial treatment regimen that included pyrazinamide. Thirty-eight inactive HBsAg carriers (35%) and 19 control subjects (20%) exhibited elevated liver enzyme levels during antituberculosis treatment (p = 0.016). Drug-induced hepatotoxicity, which was defined as a liver transaminase level of >/= 120 IU/L, occurred more frequently in HBsAg carriers (9 of 110 carriers; 8%) than in control subjects (4 of 97 control subjects; 4%), although this was not a statistically significant discrepancy (p = 0.230). More importantly, HBsAg carriers (n = 9; 8%) who received antituberculosis therapy evidenced a higher proportion of moderate-to-severe drug-induced hepatotoxicity when compared with the control subjects (n = 2; 2%; p = 0.05). Isoniazid and rifampin were reintroduced as therapy after AST/ALT levels returned to baseline values in 10 patients (6 HBsAg carriers and 4 control subjects) among the 13 patients exhibiting drug-induced hepatotoxicity, and these retrials proved to be successful in 7 patients (5 HBsAg carriers and 2 control subjects). CONCLUSIONS: Tuberculosis treatment in HBsAg-positive and HBeAg-negative inactive carriers could be pursued in the usual manner, using standard short-course regimens of isoniazid, rifampin, ethambutol, and/or pyrazinamide, with the condition that monthly liver function tests are performed. PMID- 15821210 TI - Cysteinyl leukotrienes in allergic inflammation: strategic target for therapy. AB - Systemically bioavailable leukotriene receptor antagonists (LTRAs) can reduce the essential components of allergic inflammation in allergic rhinitis (AR) and asthma by blocking cysteinyl leukotriene (CysLT) activity, resulting in a wide range of clinical effects. CysLTs, mediators, and modulators in the pathophysiology of asthma and AR are a key target for therapy because they modulate production of hemopoietic progenitor cells, survival and recruitment of eosinophils to inflamed tissue, activity of cytokines and chemokines, quantity of exhaled NO, smooth-muscle contraction, and proliferation of fibroblasts. The mechanism of action of LTRAs leads to their effects on systemic allergic inflammatory processes. PMID- 15821211 TI - Cariporide potentiates the effects of epinephrine and vasopressin by nonvascular mechanisms during closed-chest resuscitation. AB - BACKGROUND: The efficacy of vasopressor therapy during closed-chest resuscitation is limited and decreases over time. We previously reported that sodium-hydrogen exchanger isoform-1 inhibition during ventricular fibrillation (VF) using cariporide ameliorates ischemic contracture and enhances the efficacy of chest compression. We currently investigated whether cariporide could potentiate pressor responses to epinephrine and vasopressin. METHODS: VF was induced and left untreated for 12 min in two series of 16 rats each. Chest compression was then started and the depth adjusted within the initial 2 min to attain an aortic diastolic pressure between 26 and 28 mm Hg. In one series, rats received boluses of epinephrine (150 microg/kg); in the other series, rats received boluses of vasopressin (0.8 U/kg) to maintain the aortic diastolic pressure > 25 mm Hg. Within each series, rats were randomized to receive a 3 mg/kg bolus of cariporide or 0.9% NaCl immediately before starting chest compression. Defibrillation was attempted at 20 min of VF (8 min of chest compression). RESULTS: Cariporide prompted higher and more sustained coronary perfusion pressures in both the epinephrine group (37 +/- 5 mm Hg vs 29 +/- 7 mm Hg, p < 0.05) and the vasopressin group (36 +/- 5 mm Hg vs 28 +/- 6 mm Hg +/- SD, p < 0.02) even though fewer additional vasopressor doses were required. After resuscitation, cariporide treated rats had less ventricular ectopic activity, better hemodynamic function, and improved survival scores. In separate experiments, in situ perfusion of the aorta excluded a vascular-mediated effect of cariporide. CONCLUSION: Cariporide enhanced the hemodynamic efficacy of vasopressor agents and improved resuscitation outcomes probably as a result of enhanced forward blood flow without effect on the peripheral vasculature. PMID- 15821212 TI - Transforming growth factor-beta3 induces pleurodesis in rabbits and collagen production of human mesothelial cells. AB - STUDY OBJECTIVE: The aim of this study was to examined whether transforming growth factor (TGF)-beta(3) induces pleurodesis in rabbits and collagen production by human pleural mesothelial cells in vitro. DESIGN: A combined animal and in vitro study. METHODS: TGF-beta(3) was injected intrapleurally in rabbits at the following doses: 0.167 mug, 0.5 mug, 1.67 mug, and 5 mug (five rabbits per group). The rabbits were killed 14 days following injection, and pleurodesis was graded from 1 (none) to 8 (complete symphysis). Pleural mesothelial cell cultures were established from benign pleural effusions and treated with TGF-beta(3) for 48 h at the following doses: 0 (control), 5 ng/mL, 1.5 ng/mL, 5 ng/mL, and 15 ng/mL and 50 ng/mL (two independent experiments). Collagen I messenger RNA (mRNA) expression was quantified using real-time polymerase chain reaction. RESULTS: The median pleurodesis score was 5 (interquartile range [IQR], 4.5) in the 0.167-mug group, 6 (IQR, 5) in the 0.5-mug group, 8 (IQR, 1) in the 1.67-mug group, and 8 (IQR, 0.5) in the 5-mug group (p = 0.012). Higher TGF-beta(3) doses induced the production of significantly more pleural fluid than the lower doses (p = 0.005). The pleural fluid induced by higher doses contained significantly lower numbers of nucleated cells (p = 0.014) and lactate dehydrogenase levels (p = 0.013) than the pleural fluid induced by lower doses of the agent. TGF-beta(3) markedly enhanced collagen I mRNA expression by the human pleural mesothelial cells. This effect peaked at 5 ng/mL TGF-beta(3). With this dose of TGF-beta(3), the collagen I mRNA expression was increased 16-fold over control levels. CONCLUSION: TGF beta(3) causes a dose-dependent pleurodesis when administered intrapleurally in rabbits, and induces collagen messenger RNA synthesis from human pleural mesothelial cells. PMID- 15821213 TI - Decreased CCAAT/enhancer binding protein transcription factor activity in chronic bronchitis and COPD. AB - BACKGROUND: CCAAT/enhancer binding proteins (C/EBPs) are key regulators of cell differentiation and linked processes such as proliferation, apoptosis, and gene expression in several organs. C/EBPs are also central for inflammatory responses and infectious defenses, but so far little is known of their role in lung diseases. Chronic bronchitis (CB) and COPD are common smoking-associated lung diseases involving the airway epithelium. METHODS: Gelshifts were used to study C/EBP transcription factor activity in airway epithelial cells obtained by bronchial brush biopsy in four groups: healthy never-smokers (n = 10), asymptomatic smokers (n = 7), and smokers with CB and recurrent infectious exacerbations without COPD (n = 23) and with COPD (n = 13). RESULTS: C/EBP binding activity was increased 4.6-fold in airway epithelial cells of healthy smokers compared with never-smokers. In contrast, C/EBP binding activity was not increased in the epithelium of smokers with CB or COPD. C/EBP-beta was the dominant C/EBP in the airway epithelium in all groups. CONCLUSIONS: We hypothesize that this lack of increase in C/EBP-beta activity renders the epithelium incompetent of efficient regeneration and more sensitive to infection, suggesting a previously unknown role for C/EBPs in the pathogenesis of CB and COPD. PMID- 15821214 TI - Indications, timing, and techniques of tracheostomy in 152 French ICUs. AB - STUDY OBJECTIVES: To investigate the current practice of tracheostomy in French ICUs, focusing on the frequency, timing, indications, and techniques. DESIGN: A retrospective study. SETTING: A questionnaire was sent to all ICUs included in the national registry of the Societe de Reanimation de Langue Francaise. PATIENTS: All ICU patients receiving mechanical ventilation (MV) during the year preceding the survey (excluding noninvasive ventilation) were covered by the questionnaire. INTERVENTION: None. MEASUREMENTS AND RESULTS: Of the 708 ICUs invited to participate, 152 (21.5%) answered the questionnaire. Overall, 35,322 patients (median, 212 patients per unit; interquartile range [IQR], 148 to 329 patients) had received MV, including one fourth of patients for > 7 days. A median of 7.2% of patients (IQR, 2.9 to 11.1%) had undergone a tracheostomy. The most frequently reported indications for tracheostomy were prolonged MV (95%, after a median of 20 days) and failure of extubation (48%). An indication for "early" tracheostomy (ie, < 3 weeks) was considered in 68% of the ICUs, after a median time of 7 days. Physicians thought that performing tracheostomy would facilitate weaning from MV and early oral nutrition, and would improve overall patient comfort. The main disadvantages listed were tracheal complications, the aggressiveness of the procedure, and the risk of infection. Surgical techniques largely remained preferred compared to percutaneous techniques. CONCLUSIONS: There is much heterogeneity in the reported practices of tracheostomy in French ICUs. Three fourths of physicians considered that a randomized study comparing prolonged translaryngeal intubation and early tracheostomy in patients expected to receive prolonged MV would be necessary to clarify those issues. PMID- 15821215 TI - Stable patients receiving prolonged mechanical ventilation have a high alveolar burden of bacteria. AB - INTRODUCTION: In patients receiving prolonged mechanical ventilation (PMV), quantitative bronchoscopic culture has not been validated for the diagnosis of ventilator-associated pneumonia (VAP). OBJECTIVE: To measure the alveolar burden of bacteria in patients receiving PMV. SETTING: Respiratory care units of a university hospital and a long-term care facility. PATIENTS: Fourteen patients requiring PMV without clinical evidence of pneumonia. MEASUREMENTS: Quantitative culture of BAL from the right middle lobe and lingula. RESULTS: In 29 of 32 lobes, there was growth of at least one organism at > 10(4) cfu/mL. Most lobes had polymicrobial growth. CONCLUSIONS: Stable patients receiving PMV without clinical pneumonia have a high alveolar burden of bacteria. The bacterial burden in most patients exceeds the commonly accepted threshold for diagnosing VAP. The utility of quantitative bronchoscopic culture in the diagnosis of VAP in this patient population requires further study. PMID- 15821216 TI - Safety and tolerability of nonbronchoscopic lavage in ARDS. AB - STUDY OBJECTIVE: This study compared the safety profiles of bronchoscopic lavage with nonbronchoscopic lavage in the investigation of patients with acute lung injury (ALI) or ARDS. DESIGN: Single-center, randomized, cross-over study. SETTING: General ICU in the United Kingdom. PARTICIPANTS: Fourteen patients with ALI or ARDS. INTERVENTIONS: Bronchoscopic BAL and nonbronchoscopic BAL 1 h apart. MEASUREMENTS AND RESULTS: Hemodynamic and ventilatory parameters were recorded during and for 1 h following each procedure. On average, bronchoscopic lavage took longer to perform than nonbronchoscopic lavage (7 min and 6 s vs 2 min and 28 s, p < 0.001). During the procedures, bronchoscopic lavage increased heart rate and systolic BP more than nonbronchoscopic lavage (23% vs 10% [p < 0.01] and 18% vs 7% [p < 0.01]). Three patients had ST-segment depression during bronchoscopic, and one patient had ST-segment depression during nonbronchoscopic lavage (p = 0.298). Bronchoscopic lavage reduced minute ventilation by 63 +/- 17.3%, while nonbronchoscopic lavage only reduced it by 36 +/- 21.9% (p < 0.001). Paco(2) rose more after bronchoscopic lavage than after nonbronchoscopic lavage. CONCLUSION: Nonbronchoscopic lavage is associated with less marked physiologic derangements than bronchoscopic lavage. Further studies are required to validate the hypothesis that nonbronchoscopic lavage may be safer in patients with unstable coronary heart disease or head injury/raised intracranial pressure who are at risk from unpredictable fluctuations in hemodynamic and ventilatory profiles. PMID- 15821217 TI - Long-term performance of a simple algorithm for early discharge after ruling out acute coronary syndrome: a prospective multicenter trial. AB - STUDY OBJECTIVES: To test the short- and long-term performance of a simple algorithm to detect or rule out acute coronary artery disease (CAD) in patients referred with acute chest pain (CP). DESIGN: Prospective, multicenter study with 30-day and 1-year follow-ups. SETTING: Emergency and coronary care units of two university hospitals and four affiliated teaching hospitals in a suburban region of northern Switzerland. PATIENTS: One hundred sixty-one consecutive patients referred with acute CP with a high CAD risk profile underwent exercise testing (ET) within 24 h if results of two troponin tests and ECG remained normal within 6 h. The patients were discharged if ET results were negative or underwent angiography if the results were positive. MEASUREMENTS AND RESULTS: The primary end point was survival free of death, myocardial infarction (MI), and acute coronary syndrome, as well as the correct diagnosis of CAD in need of revascularization. Failure of the algorithm included events not predicted and false-positive ET results. Symptom-limited ET was performed in 142 patients, and stress imaging was performed in 17 patients. Discharge within 24 h was possible in 76%. Angiography in 21 patients showed no CAD in 4 patients, whereas revascularization was necessary in 17 patients. During follow-up, three patients had an MI (one fatal) and two patients had unstable angina, for an event rate of 3.1%/yr. Overall sensitivity of the algorithm to detect CAD was 71%, whereby six of seven false-positive ET results were due to exercise-induced CP without ECG changes. More importantly, the negative predictive value was 96.4%, indicating that this simple algorithm failed to identify only 3.6% of patients at risk for future events. CONCLUSIONS: Discharge within 24 h after onset of acute CP in high risk CAD patients is safe, irrespective of CAD history, if ECG findings remain unchanged and troponin test results are normal within 6 h, and if results of adequate ET are negative. PMID- 15821218 TI - A pictorial review of coronary artery bypass grafts at multidetector row CT. AB - Multidetector row CT scanners with submillimeter spatial resolution and high temporal resolution are now available and are increasingly used for noninvasive assessment of vascular disease including coronary arteries and grafts. The entire heart and graft course can be scanned within a single breath-hold, and contrast enhanced images are reconstructed through retrospective ECG gating. In this pictorial review, we describe the CT findings of the most commonly used coronary artery bypass grafts on both axial images and two-dimensional and three dimensional images providing a correlation with conventional coronary angiography. PMID- 15821219 TI - Endoscopic drainage of lung abscesses: technique and outcome. AB - BACKGROUND: Lung abscesses commonly respond well to antibiotic therapy. In patients in whom conventional therapy fails, either percutaneous catheter drainage or surgical resection are usually considered, but are frequently problematic. This study describes our experience with endoscopic lung abscess drainage in patients in whom antibiotic therapy fails. METHODS: Patients in whom antibiotic therapy for lung abscess (enlarging cavity or lack of improvement of clinical status) was unsuccessful were considered candidates if an airway connection to the cavity was present. Treatment decisions were made in a multidisciplinary chest conference. Pigtail catheters were placed via a guidewire approach into the cavities. The abscesses were flushed twice daily with gentamycin solution. If fungal infection was suspected, once-daily amphotericin B was added to the regimen. RESULTS: Forty-two patients, from January 2000 to May 2002 (17 woman and 25 men) were included in this study (mean age, 48.9 years). Catheter placement was successful in 38 patients and led to successful therapy after a mean of 6.2 days of treatment (range, 3 to 21 days). Two patients required transient ventilation after catheter placement; there were no other complications. CONCLUSIONS: Endoscopic lung abscess drainage in selected patients in whom antibiotic therapy fails is feasible and successful in experienced hands. This treatment represents an additional option for the chest physician other than percutaneous catheter drainage or surgical resection. PMID- 15821220 TI - Reducing maintenance and repair costs in an interventional pulmonology program. AB - BACKGROUND: In the current economic climate, hospitals and academic institutions demand that medical departments function in an efficient and cost-effective manner. Detailed business plans are necessary to build new clinical programs, and institutions have learned that new programs are associated with significant costs for purchasing and maintaining equipment. We report our experience with repairs to equipment before and after starting our interventional pulmonary (IP) program, and with the effect of an educational program on reducing these costs. METHODS: We retrospectively studied the costs of equipment repair in the 3 years preceding and in the 5 years following the development of an IP program in our institution, a university-based tertiary referral center. We also studied the effect of an educational program that was designed to enhance the skills of physicians and technical staff in handling the equipment. RESULTS: The cost of repairs to the equipment during the 3 years prior to the development of the IP program was $42 (US dollars) per procedure. In the initial 3 years following the start of the IP program, the yearly average cost rose 21% to $51 per procedure. After the introduction of the educational program, the yearly repair costs decreased by 84% to $8 per procedure. Based on our experience, we estimate that a reasonable budget for the cost of repairs is $50 per procedure. CONCLUSIONS: An educational program was effective in dramatically decreasing the costs of equipment repair after initiating an IP program. This is the first study to offer budgetary guidelines for equipment repair in an IP program and to demonstrate that an educational program can effectively reduce costs. PMID- 15821221 TI - The influence of diagnostic bronchoscopy on clinical outcomes comparing adult autologous and allogeneic bone marrow transplant patients. AB - STUDY OBJECTIVES: To review our experience with diagnostic bronchoscopy in the evaluation of pulmonary infiltrates in adult hematopoietic stem cell transplantation (HSCT) recipients in the era of Pneumocystis prophylaxis and cytomegalovirus antigen testing. The study focused on diagnostic yields and the influence of bronchoscopic findings on pharmacologic therapy and mortality, comparing allogeneic (allo) HSCT patients to autologous (auto) HSCT patients. DESIGN: Case series review. SETTING: Tertiary care academic urban medical centers. PATIENTS: All adult allo-HSCT and auto-HSCT patients undergoing bronchoscopy for the evaluation of pulmonary infiltrates from January 1997 to September 2001. MEASUREMENTS AND RESULTS: The review identified 169 bronchoscopies that had been performed on HSCT patients, representing 12.5% of all HSCT patients (allo-HSCT patients, 125 bronchoscopies; auto-HSCT patients, 44 bronchoscopies). Bronchoscopy was requested more often in allo-HSCT patients (18.7%) compared to auto-HSCT patients (6.6%). Findings at bronchoscopy provided a specific diagnosis more frequently in allo-HSCT patients (50%) compared to auto HSCT patients (34%). For both allo-HSCT and auto-HSCT patients, most diagnoses were obtained by BAL alone, whereas transbronchial biopsy (TBBx) provided additional specific information in < 10% of cases. For select patients (n = 27), surgical lung biopsy following bronchoscopy provided unique diagnoses in 47 to 50% of cases. Information from bronchoscopy influenced clinical decisions more often in allo-HSCT patients (50%) than in auto-HSCT patients (36%), and allowed for the discontinuation or addition of antimicrobial, corticosteroid, or antineoplastic agents to treatment. Complications from bronchoscopy occurred in 9% of all HSCT patients (n = 15), and were associated with higher in-hospital mortality rates in allo-HSCT patients (82%; n = 9) compared to auto-HSCT patients (50%; n = 2). The overall in-hospital mortality rates for allo-HSCT and auto-HSCT patients having bronchoscopy was similar (38% vs 27%, respectively; p = 0.25), and establishing a specific diagnosis by bronchoscopy did not improve the in hospital mortality rate for allo-HSCT or auto-HSCT patients. CONCLUSIONS: Bronchoscopy may provide clinically useful information in the evaluation of adult allo-HSCT and auto-HSCT recipients with pulmonary infiltrates. The results of testing BAL fluid samples alone suggested an etiology in most cases, whereas the findings of TBBx provided unique diagnoses infrequently. Further studies are warranted to improve the utility of diagnostic bronchoscopy in the evaluation of HSCT patients. PMID- 15821222 TI - Airway management of the critically ill patient: rapid-sequence intubation. AB - Advances in emergency airway management have allowed intensivists to use intubation techniques that were once the province of anesthesiology and were confined to the operating room. Appropriate rapid-sequence intubation (RSI) with the use of neuromuscular blocking agents, induction drugs, and adjunctive medications in a standardized approach improves clinical outcomes for select patients who require intubation. However, many physicians who work in the ICU have insufficient experience with these techniques to adopt them for routine use. The purpose of this article is to review airway management in the critically ill adult with an emphasis on airway assessment, algorithmic approaches, and RSI. PMID- 15821223 TI - Effects of N-acetylcysteine on microalbuminuria and organ failure in acute severe sepsis: results of a pilot study. AB - STUDY OBJECTIVE: The level of microalbuminuria is thought to reflect the severity of inflammation-induced systemic vascular permeability and may have prognostic value with regard to organ dysfunction and survival. N-acetylcysteine (NAC) has been shown to decrease capillary leakage in experimental sepsis. The present study investigated the effect of early treatment with NAC on microalbuminuria and organ dysfunction in severe clinical sepsis. DESIGN: Prospective, randomized, placebo-controlled study. SETTING: A 24-bed multidisciplinary ICU in a university teaching hospital. PATIENTS: Thirty-five patients included within 4 h of fulfilling consensus criteria of severe sepsis. INTERVENTIONS: Patients were randomly assigned to receive either NAC (continuous infusion starting with 50 mg/kg/4 h followed by 100 mg/kg/24 h for 44 h; n = 18) or placebo (n = 17) in addition to standard therapy. MEASUREMENTS AND RESULTS: Urine samples for measurement of microalbuminuria/creatinine ratio (MACR) were collected on inclusion and after 4 h, 24 h, and 48 h. Severity of illness and degree of organ failure were determined by using, respectively, the APACHE (acute physiology and chronic health evaluation) II score and the sequential organ failure assessment (SOFA) score. The MACR did not differ over time between the placebo- and the NAC treated groups. SOFA scores were comparable between both treatment groups at baseline (6.2 +/- 3.9 vs 6.5 +/- 2.7, NAC vs placebo; p = 0.6) and increased during treatment in the NAC-treated patients but not in the placebo group (7.9 +/ 3.7 vs 5.9 +/- 2.5, p = 0.09 and 7.7 +/- 3.8 vs 5.1 +/- 2.1, p < 0.05; NAC vs placebo, respectively, at 24 h and at 48 h). The cardiovascular SOFA score progressively increased during NAC treatment, reaching higher values as compared to time-matched scores in the placebo group. CONCLUSIONS: Early NAC administration does not influence the course of MACR in severe clinical sepsis, suggesting that NAC might not attenuate endothelial damage in this condition. NAC treatment even aggravated sepsis-induced organ failure, in particular cardiovascular failure. PMID- 15821224 TI - A pilot study of the safety and efficacy of tobramycin solution for inhalation in patients with severe bronchiectasis. AB - STUDY OBJECTIVE: To evaluate the efficacy and safety of tobramycin solution for inhalation (TSI) in patients with severe bronchiectasis. DESIGN: Open-label clinical trial consisting of three treatment cycles (14 days of drug therapy, and 14 days off drug) and an additional 40-week follow-up by chart review. SETTING: Nine clinical sites throughout the United States. SUBJECTS: Forty-one adult patients (>/= 18 years old) with diffuse bronchiectasis affecting two or more lung segments and a history of Pseudomonas aeruginosa infection. INTERVENTIONS: TSI, 300 mg tobramycin per dose bid. MEASUREMENTS AND RESULTS: During the 12-week treatment period, significant improvements (reduction of 1.5 U [p = 0.006]) occurred in mean pulmonary total symptom severity score, a composite score that assesses the severity of cough, shortness of breath, sputum production, fatigue, and wheezing. Significant improvements (reduction of 9.8 U [p < 0.001]) were also observed in St. George Respiratory Questionnaire scores, which measure health related quality of life. Eradication or presumed eradication of P aeruginosa occurred in 6 of 27 evaluable subjects (22.2%). Tobramycin-resistant P aeruginosa developed in two subjects (minimal inhibitory concentration >/= 16 microg/mL). Ten subjects withdrew from the study due to adverse events; in nine of these subjects, adverse events were considered probably or possibly related to treatment. The most common adverse events were cough, wheezing, and dyspnea. CONCLUSIONS: TSI therapy resulted in significant improvements in respiratory symptoms and health-related quality of life in subjects with severe bronchiectasis, but some subjects did not tolerate TSI therapy. Bronchiectasis patients receiving this therapy should be monitored for signs of intolerance. PMID- 15821225 TI - Video-assisted thoracoscopic surgery in the treatment of complicated parapneumonic effusions or empyemas: outcome of 234 patients. AB - STUDY OBJECTIVE: To review our experience in treatment of complicated parapneumonic effusion and pleural empyema by video-assisted thoracoscopic surgery (VATS). DESIGN: Retrospective chart review. SETTING: Taiwanese medical centers. PATIENTS: A total of 234 patients (108 women, 126 men; median age, 51 years; range, 0.75 to 84 years) underwent procedures for parapneumonic effusion (145 patients) or pleural empyema (89 patients) between May 1995 and December 2003. All patients had chest radiographs, and 188 patients (80.3%) underwent preoperative CT or sonography. More than 85% (200 patients) received preoperative diagnostic or therapeutic thoracentesis, tube thoracostomy, or fibrinolytics. Indications for VATS included empyema refractory to medical control or peel or multiloculated exudates per CT and chest tapping. INTERVENTIONS: Septal lysis and debridement irrigation through one port (31 patients, 13.2%), decortication and debridement through two or three ports (179 patients, 76.5%), or rib resection or larger utility incision for decortication and drainage (24 patients, 10.3%). RESULTS: Mean +/- SD procedural time was 64.3 +/- 22.5 min (range, 26 to 244 min). Sixteen patients (6.8%) needed further surgery for empyema (9 patients required open drainage or thoracoplasty, and 7 patients needed redecortication or repair of bronchopleural fistula). There were no intraoperative deaths and only eight (3.4%) perioperative deaths (< 30 days), which were mostly unrelated to surgery. Of the 234 patients, 202 patients (86.3%) achieved satisfactory results with VATS treatment. Patients requiring open decortication or repeat procedures (40 patients) had a longer mean duration of preoperative symptoms, longer mean duration of preoperative hospitalization, and a higher ratio of pleural empyema (vs complicated parapneumonic effusion) than patients undergoing simple VATS. CONCLUSIONS: VATS is safe and effective for treatment of complicated parapneumonic effusion and pleural empyema. Earlier intervention with VATS can produce better clinical results. A prospective study should be done to identify optimal timing and settings for VATS treatment for both complicated parapneumonic effusion and pleural empyema. PMID- 15821226 TI - Lung masses in a 70-year-old man. PMID- 15821227 TI - Dyspnea, hemoptysis, and perihilar infiltrates in a 35-year-old man. PMID- 15821228 TI - Orthopnea in a 75-year-old man after cardiac catheterization. PMID- 15821229 TI - Simvastatin treatment of pulmonary hypertension: an observational case series. AB - BACKGROUND: Statins confer cardiovascular benefits beyond the reduction of serum cholesterol through antiproliferative and antiinflammatory mechanisms and induction of endothelial nitric oxide expression. In pneumonectomized rats injected with monocrotaline, simvastatin reversed established pulmonary hypertension and conferred a 100% survival advantage. STUDY OBJECTIVES: To evaluate the safety and efficacy of simvastatin for treatment of patients with pulmonary arterial hypertension (PAH). DESIGN: Open-label observational study performed at Stanford University Medical Center. Sixteen patients with primary and secondary causes of PAH, World Health Organization (WHO) classes I (n = 2), II (n = 4), III (n = 3), IV (n = 7), are described. Simvastatin was prescribed at 20 to 80 mg/d and continued in the absence of adverse effects. MEASUREMENTS AND RESULTS: Serial measurements of 6-min walk (6MW) performance, hemodynamics, and echocardiographic estimates of right ventricular systolic pressures (RVSPs) were recorded on each patient. Simvastatin treatment was not associated with hepatic dysfunction, muscle necrosis, or other adverse events. Individual patients demonstrated improvements in 6MW performance, improvements in cardiac output, or decreases in RVSP that may be attributable to simvastatin treatment. Overall, the rate of disease progression appeared to be attenuated, and WHO class IV patients demonstrated improved survival. CONCLUSIONS: Simvastatin treatment appears safe in patients with PAH. PMID- 15821230 TI - Peripheral arterial embolism due to a left ventricular diverticulum in a young adult. AB - A 32-year-old man was admitted to the emergency department of our hospital after experiencing a peripheral arterial embolism. Investigation of the possible embolic sources in an otherwise asymptomatic patient revealed the existence of a left ventricular diverticulum. The left ventricular diverticulum is a rare congenital anomaly, either isolated or as a part of a syndrome including other congenital malformations. The treatment of choice, especially in symptomatic patients, is surgical resection, while in asymptomatic patients anticoagulation therapy is indicated. PMID- 15821231 TI - Exercise testing in Wolff-Parkinson-White syndrome: case report with ECG and literature review. AB - ECG changes during exercise stress testing, such as false-positive ST-segment depression and disappearance of the delta wave, are reported in patients with the Wolff-Parkinson-White (WPW) pattern. We present a case of exercise testing in a 53-year-old man with WPW syndrome with ischemic-appearing ECG changes and normal nuclear stress perfusion study findings who was thought to be at clinically low risk for having significant coronary disease. A literature review is discussed. Although ST-segment depression typical for ischemia occurs in half of the patients in whom WPW syndrome is reported, exercise testing is still an important tool in their evaluation. Data other than ECG response can be interpreted in the context of clinical history and physical examination findings to stratify the risk of coronary disease. Complete and sudden disappearance of the delta wave has been seen during exercise in 20% of patients with WPW syndrome and can identify those who are at low risk for sudden arrhythmic death. PMID- 15821232 TI - Right ventricular dysfunction. PMID- 15821233 TI - Beneficial antiinflammatory effects of leukotriene receptor antagonists in asthma. PMID- 15821234 TI - Interpretation of transpulmonary thermodilution curves. PMID- 15821235 TI - Effects of leukotriene receptor antagonists on vascular endothelial growth factor levels in asthma. PMID- 15821236 TI - Atrial fibrillation, atrial flutter, or both after pulmonary transplantation. PMID- 15821238 TI - Physician interaction with internet pharmacies is risky. PMID- 15821239 TI - There are other ways to manage spinal muscular atrophy type 1. PMID- 15821240 TI - Apoptosis of circulating neutrophils in COPD patients. PMID- 15821241 TI - SNAP technology and sleep apnea. PMID- 15821242 TI - D-dimer in the diagnostic workup of suspected pulmonary embolism: additional clinical value of pretest probability. PMID- 15821244 TI - Pathological and clinical response of a primary chemotherapy regimen combining vinorelbine, epirubicin, and paclitaxel as neoadjuvant treatment in patients with operable breast cancer. AB - This phase II study investigated the efficacy and tolerability of a primary chemotherapy regimen combining vinorelbine, epirubicin, and paclitaxel (VEP protocol) in women with stage II/III operable breast cancer. Patients (n = 50) were treated with six cycles of VEP according to the following schedule: vinorelbine (Navelbine); Pierre Fabre, Boulogne, France; http://www.pierre fabre.com) 20 mg/m2, epirubicin (Farmorubicin; Pharmacia, New York, NY; http://www.pnu.com) 35 mg/m2 given on days 1 and 8, paclitaxel (Taxol; Bristol Myers Squibb, New York, NY; http://www.bmsoncology.com) 175 mg/m2 given on day 9, and G-CSF 5 mg/kg/day given on days 10-20 of a 21-day cycle, followed by surgery and radiotherapy. After six cycles of VEP, the pathological response rate (pCR) in breast was confirmed in six patients (12%; 95% confidence interval [CI]: 3 21)) using Chevallier's classification and in nine patients (18%; 95% CI: 7.4 28.6) using Sataloff's classification. The clinical response rate was 42% (95% CI: 28.3-55.7), including 26% complete responses. Breast conservation was achieved in 68% of patients. After a median follow-up of 48 months (range, 34-62 months), 16 relapses were observed. The overall and disease-free survivals at 5 years were 54.1% (95% CI: 40.3-67.9) and 38% (95% CI: 24.1-51.9), respectively. The principal toxicities of VEP were grade 3/4 neutropenia observed in 30% of patients and grade 3 anemia observed in 12% of patients. There was no case of severe cardiac toxicity, thrombocytopenia, or any other serious adverse events. In conclusion, whereas this regimen was relatively well tolerated, it appears inferior to other regimens and its use is not recommended. PMID- 15821245 TI - Critical evaluation of current treatments in metastatic colorectal cancer. AB - Fluorouracil (FU) has been the mainstay of treatment for metastatic colorectal cancer (mCRC) for many years. However, in recent years, newer chemotherapeutic agents, particularly irinotecan (Campostar; Pfizer Pharmaceuticals, New York, NY, http://www.pfizer.com) and more recently oxaliplatin (Eloxatin; Sanofi-Aventis Inc., New York, NY, http://www.sanofi-aventis.com), have been shown to improve survival in combination with FU-based therapies. These agents were therefore incorporated into first- and second-line treatment strategies. The development of targeted agents that are tumor specific with better toxicity profiles than chemotherapeutic agents has widened the spectrum of therapies for this disease. The U.S. Food and Drug Administration (FDA) recently approved two targeted agents for treating mCRC: an antivascular endothelial growth factor monoclonal antibody (mAb), bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA, http://www.gene.com), in combination with first-line 5-FU-based chemotherapy regimens and the human epidermal growth factor receptor (HER-1/EGFR)-targeted mAb cetuximab (Erbitux; ImClone Systems, Inc., New York, NY, http://www.imclone.com) as monotherapy or in combination with irinotecan as second-line therapy in refractory cancer. These newer, more effective agents are improving clinical outcome for patients with mCRC. However, as the number of agents has increased, choosing the most effective treatment strategy has become increasingly complex. This review discusses the role of the individual agents in the treatment of mCRC and identifies the most effective regimens. PMID- 15821246 TI - Preoperative assessment of surgical risk in oncogeriatric patients. AB - Cancer is a prevalent disease in our aging population; however, few oncologists are familiar with caring for oncogeriatric patients. Surgery is presently the treatment of choice for most solid tumors, but it is frequently delivered in a suboptimal way in this patient subsetting. Undertreatment is often justified with the concern of an unsustainable toxicity, while overtreatment can be related to the lack of knowledge in optimizing preoperative risk assessment. To draw new light on this issue, several surgeons presented their series, providing hard evidence that surgical options can be offered to the elderly with cancer, with only a limited postoperative mortality and morbidity. As it is likely that much of these data suffer from selection bias, we concentrated on Comprehensive Geriatric Assessment (CGA), which can add substantial information on the functional assessment of elderly cancer patients. A validated instrument such as the CGA allows a comparison of series, predicting short-term surgical outcomes more precisely, and offers appropriate information when consenting elderly patients. Preoperative Assessment of Cancer in the Elderly is a prospective international study conceived and launched to outline the fitness of elderly surgical patients with malignant tumors. This paper reports on preliminary results and analysis from the ongoing study. PMID- 15821247 TI - Liver transplantation for malignancy. AB - Liver transplantation for hepatic malignancies has emerged from an exotic and desperate approach to a well-documented and proven treatment modality for these unfortunate patients. However, early unsatisfactory results emphasized that only a highly selected patient population would benefit from transplantation. Currently, <10% of all liver transplants performed are for hepatocellular cancer (HCC). There is no controversy that hepatoblastoma is an excellent indication in pediatric patients with unresectable tumors. Similarly, liver transplantation for HCC in the adult population yields good results for patients whose tumor masses do not exceed the Milan criteria. It remains to be determined whether patients with more extensive tumors can be reliably selected to benefit from the procedure. Adjunctive procedures like radiofrequency ablation, chemoembolization, or cryotherapy might be indicated to limit tumor progression for patients on waiting lists. Epitheloid hemangioendothelioma is also an appropriate indication for liver transplantation, unlike angiosarcoma. Metastatic liver disease is not an indication for liver transplantation, with the exception of cases in which the primary is a neuroendocrine tumor, for which liver transplantation can result in long-term survival and even cure in a number of patients. And finally, while gallbladder cancers are never an indication for liver transplantation, rare cases of cholangiocellular cancer might qualify if aggressive combination therapies, including chemotherapy and radiotherapy followed by OLT, are carried through. Survival in these selected patients can approach that for patients with cholestatic liver disease. PMID- 15821248 TI - Three emerging new drugs for NSCLC: pemetrexed, bortezomib, and cetuximab. AB - Despite advances made in cytotoxic chemotherapy, the prognosis for patients with non-small cell lung cancer (NSCLC) continues to be poor. New, more effective drugs must be identified and developed to improve the outcome of these patients. Three drugs with promising activity in NSCLC are pemetrexed (Alimta; Eli Lilly and Company, Indianapolis, IN, http://www.lilly.com), bortezomib (Velcade; Millennium Pharmaceuticals, Inc., Cambridge, MA, http://www.mlnm.com), and cetuximab (Erbitux; ImClone Systems, Inc., New York, NY, http://www.imclone.com). Pemetrexed inhibits thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyl transferase, enzymes necessary for purine and pyrimidine synthesis, thus causing cell-cycle arrest in the S phase. Bortezomib, a proteasome inhibitor, interferes with the cytosolic protein degradation machinery, namely the ubiquitin-proteasome complex, causing breakdown of cell cycle regulators and cell-cycle arrest. Cetuximab is a chimeric mouse-human antibody that inhibits ligand-dependent activation of the epidermal growth factor receptor, resulting in receptor internalization and inhibition of downstream pathways that, in turn, causes cell growth and progression. All three drugs are approved for different tumor types, and studies defining their role in NSCLC are under way. PMID- 15821249 TI - T-cell lymphoma of the rectum in a patient with AIDS and hepatitis C: a case report and discussion. AB - Primary T-cell non-Hodgkin's lymphoma (NHL) occurring in the context of acquired immune deficiency syndrome (AIDS) is uncommon. Here, we report and discuss such a case presenting in the rectum, and review relevant literature. Although typical in some respects, the case is, in other ways, somewhat unusual for an AIDS related NHL (ARL); ARL tends to be B cell and advanced stage and our case was T cell and stage IE. In addition, the patient suffered from concomitant cirrhosis related to hepatitis C. Chemotherapeutic options for ARL were limited early in the AIDS epidemic due to poor tolerability. Although this has largely been mitigated by the advent of highly active antiretroviral therapy, our patient eventually suffered complications of chemotherapy, apparently related more to his liver disease than to either his lymphoma or AIDS, that ultimately brought about his demise. PMID- 15821250 TI - Thrombotic thrombocytopenic purpura secondary to an occult adenocarcinoma. PMID- 15821251 TI - Wrong primer for rat angiotensinogen mRNA. PMID- 15821252 TI - Endothelial cell dysfunction: can't live with it, how to live without it. AB - Endothelial cell dysfunction is emerging as an ultimate culprit for diverse cardiovascular diseases and cardiovascular complications of chronic renal diseases, yet the definition of this new syndrome, its pathophysiology, and therapy remain poorly defined. Here, I summarize some molecular mechanisms leading from hyperhomocystinemia, elevated asymmetric dimethylarginine, and advanced glycolation end product-modified protein level to the proatherogenic, prothrombogenic, and proinflammatory endothelial phenotype and offer a model of endothelial dysfunction based on the interconnectedness of diverse functions. Finally, several therapeutic strategies to prevent and correct endothelial dysfunction are discussed in the light of uncertainty of their action modulated by the endothelial dysfunction per se. PMID- 15821253 TI - Urea and urine concentrating ability: new insights from studies in mice. AB - Urea is the most abundant solute in the urine in humans (on a Western-type diet) and laboratory rodents. It is far more concentrated in the urine than in plasma and extracellular fluids. This concentration depends on the accumulation of urea in the renal medulla, permitted by an intrarenal recycling of urea among collecting ducts, vasa recta and thin descending limbs, all equipped with specialized, facilitated urea transporters (UTs) (UT-A1 and 3, UT-B, and UT-A2, respectively). UT-B null mice have been recently generated by targeted gene deletion. This review describes 1) the renal handling of urea by the mammalian kidney; 2) the consequences of UT-B deletion on urinary concentrating ability; and 3) species differences among mice, rats, and humans related to their very different body size and metabolic rate, leading to considerably larger needs to excrete and to concentrate urea in smaller species (urea excretion per unit body weight in mice is 5 times that in rats and 23 times that in humans). UT-B null mice have a normal glomerular filtration rate but moderately reduced urea clearance. They exhibit a 30% reduction in urine concentrating ability with a more severe defect in the capacity to concentrate urea (50%) than other solutes, despite a twofold enhanced expression of UT-A2. The urea content of the medulla is reduced by half, whereas that of chloride is almost normal. When given an acute urea load, UT-B null mice are unable to raise their urinary osmolality, urine urea concentration (Uurea), and the concentration of non-urea solutes, as do wild-type mice. When fed diets with progressively increasing protein content (10, 20, and 40%), they cannot prevent a much larger increase in plasma urea than wild-type mice because they cannot raise Uurea. In both wild-type and UT-B null mice, urea clearance was higher than creatinine clearance, suggesting the possibility that urea could be secreted in the mouse kidney, thus allowing more efficient excretion of the disproportionately high urea load. On the whole, studies in UT-B null mice suggest that recycling of urea by countercurrent exchange in medullary vessels plays a more crucial role in the overall capacity to concentrate urine than its recycling in the loops of Henle. PMID- 15821254 TI - Transcriptional control of renal collecting duct development. PMID- 15821255 TI - Physiological actions of renal collecting duct endothelin. PMID- 15821256 TI - Tales from the deep: reeling in renal failure. PMID- 15821257 TI - Segmental expression of Notch and Hairy genes in nephrogenesis. AB - Notch signaling pathway genes are required for nephrogenesis, raising the possibility that Notch effector Hairy-related genes should also control nephron formation. We performed in situ hybridization of Hairy transcription factors with segment-specific lectins and/or antibodies during early nephrogenesis to identify their possible roles in segment identity of the nephron. We found that among all of Notch downstream Hairy genes, only Hes1, Hes5, Hey1, and HeyL were expressed in a segment-specific manner in early nephrons and their expression pattern changed dynamically during metanephric development. Based on these patterns of expression, it was possible to propose a pairwise association of specific ligand and receptor and to suggest that the effector of this association is one of the Hairy transcription factors. We found that Hes5 is specifically expressed in the anlage of the loop of Henle, suggesting that it might be involved in the determination of its cell identity. We also examined the morphological appearance of kidneys from mice where the Hes1 or Hes5 genes were deleted and found that at least at the gross morphological level, there was little difference from wild type kidneys. Because Hairy genes associate with other transcription factors to exert their effect, it is necessary to examine a more complete array of genetic deletions before a conclusion can be reached regarding their role in kidney development. These studies provide the basis for the future development of strategies to examine the role of individual effector molecules in the determination of the differentiation pattern of the nephron. PMID- 15821258 TI - Intramembranous absorption rate is unaffected by changes in amniotic fluid composition. AB - Experiments were performed to determine the effect of amniotic fluid dilution on the rate of intramembranous absorption. Seven fetal sheep at 118 days gestation were instrumented with a shunt between the trachea and esophagus and arterial and venous vascular catheters. In addition, the urachus of the fetal bladder was ligated, and a catheter was placed in the bladder. Ligation of the urachus does not interfere with urine flow into the amnion. After 5 days of recovery, fetuses were randomly assigned to one of two protocols; all fetuses completed both protocols. In the fetuses in the control period, continuous urine flow measurement was begun. In the fetuses assigned to the isovolumic dilution protocol, continuous urine flow measurement was also begun and, in addition, amniotic fluid was continually exchanged with lactated Ringer solution on an isovolumic basis. After 3-4 days, fetal blood pressures and amniotic fluid volumes were determined. Amniotic fluid volumes were determined by drainage. Each fetus was then assigned to the remaining protocol. The presence of the tracheal esophageal shunt and the ligation of the urachus allowed the rate of intramembranous absorption to be calculated. Isovolumic exchange showed no effect on fetal vascular pressures, blood-gas values, or urine production. We could demonstrate no effect of isovolumic dilution of amniotic fluid on its volume. However, we were able to demonstrate an inverse relationship between amniotic fluid volume and intramembranous absorption (P < 0.02). PMID- 15821259 TI - Superoxide enhances Na-K-2Cl cotransporter activity in the thick ascending limb. AB - Superoxide (O2-) enhances Na reabsorption by the thick ascending limb (THAL). Na absorption in this segment involves the Na-K-2Cl cotransporter, K channel, and Na K-ATPase. We hypothesized that O2- stimulates NaCl absorption primarily by enhancing Na-K-2Cl cotransport. First, we measured steady-state intracellular Na (Nai) and chloride (Cli). Xanthine oxidase (XO; 0.75 mU/ml) and hypoxanthine (HX; 0.125 mM) were added to the bath to increase O2-. During the control period, Nai was 12.2 +/- 1.9 mM. After treatment with O2-, it rose to 20.9 +/- 3.3 mM, a 71% increase (P < 0.01). Cli also increased (P < 0.01). Neither XO nor HX alone had a significant effect on Nai or Cli. Next, we tested cotransport activity by measuring the initial rate of increase in Nai caused by changing luminal Na-Cl-K from 50/0/0 to 140/134/4 mM. During the control period, the initial rate of increase was 0.13 +/- 0.02 arbitrary units (AU)/min. After treatment with O2-, it was 0.22 +/- 0.04 AU/min (P < 0.025), a 69% increase. Neither XO nor HX alone had a significant effect. Furosemide completely blocked the increase in intracellular Na in the control and O2- treatment periods. Next, we studied K channel activity by measuring the depolarization caused by increasing luminal K from 1 to 25 mM using a voltage-sensitive dye. During the control period, maximum depolarization was 7.31 +/- 0.77 AU. After O2- treatment, it was 6.18 +/- 0.90 AU (P < 0.05), a 15% decrease. Finally, we assessed the effects of O2- on Na-K-ATPase activity in THAL suspensions by measuring ATP hydrolysis. Vmax and K1/2 for Na were not affected by O2-. We concluded that O2- stimulates THAL NaCl absorption primarily by enhancing Na entry via Na-K-2Cl cotransport. PMID- 15821260 TI - A metamemory perspective on odor naming and identification. AB - A metacognitive perspective is utilized to elucidate why it is so difficult to name common odors and what characterizes the subjective knowledge people have about their actual odor knowledge. Odor-naming failures are often accompanied by strong feelings of knowing (FOK) or feelings of imminent retrieval of what it is that smells. The paper's two experiments investigate FOK judgements and tip of the tongue (TOT) experiences for odor and person names. The data indicate that our inability to correctly name odors are typically not due to the often proposed uniquely poor association between odors and their proper names, but rather due to failures to identify the odors, that is, failures to know 'what it is'. It was also found that (i) TOT experiences are very unusual for odor names and more so than for person names; (ii) FOK judgements about odor names are significantly less predictive of later retrieval than equivalent judgements about names of persons; (iii) FOK judgements were highly correlated with the familiarity of the cue (odor or picture of famous person), rendering some support for the idea that FOK judgements are based on the perceived familiarity of the cue triggering the FOK; and (iv) the idea that FOK judgements are based on the amount of available information about the sought-for memory (accessibility theory) was also supported. PMID- 15821261 TI - Pharmacokinetics of antiviral agents for the treatment of cytomegalovirus infection. AB - PURPOSE: The mechanism of action, pharmacokinetics, and other advantages and disadvantages of ganciclovir and valganciclovir are discussed. SUMMARY: Shortcomings of oral ganciclovir include low bioavailability, large pill burden, patient nonadherence, and the emergence of resistance. Valganciclovir, an oral prodrug of ganciclovir, has a nearly tenfold greater absolute bioavailability than ganciclovir. Dosage adjustment is required for both drugs in patients with renal impairment. CONCLUSION: The pharmacokinetic profile of valganciclovir offers significant advantages for its use in cytomegalovirus (CMV) prophylaxis. PMID- 15821262 TI - Overview of transplant mycology. AB - PURPOSE: The epidemiology, clinical manifestation, diagnosis, and management of invasive mycoses in transplant patients are described. SUMMARY: The incidence of and mortality from invasive mycoses have increased in transplant patients. Candida and Aspergillus are the two most common fungal pathogens in this patient population. The use of immunosuppressive and myeloablative therapies and other factors increases the risk of invasive mycoses. A high index of suspicion and clinical clues are needed for the diagnosis of Candida and Aspergillus infections because reliable diagnostic techniques are not available and the patient presentation is nonspecific. Targeted prophylaxis for patients at high risk for morbidity and mortality and early, aggressive treatment using broad-spectrum antifungal agents are recommended. Prophylaxis using itraconazole reduces the risk of treatment failure due to resistant pathogens compared with fluconazole. Voriconazole might prove advantageous compared with fluconazole and itraconazole for prophylaxis because of its extended spectrum and predictable blood levels. Caspofungin seems as effective as and less toxic than amphotericin B for this use. Voriconazole is more effective than amphotericin B for the treatment of aspergillosis. Caspofungin is comparable to voriconazole in efficacy as salvage treatment after failure to respond to other antifungal agents. CONCLUSION: Caspofungin plus voriconazole and other combination therapies often are used for the treatment of aspergillosis yet data to support such use are yet to be produced. The investigational agents posaconazole, micafungin, and anidulafungin appear promising as salvage treatment of various invasive mycoses. PMID- 15821265 TI - Prophylaxis of cytomegalovirus disease in transplant patients. AB - PURPOSE: A historical perspective and the epidemiology, pathogenesis, and typical patient presentation of cytomegalovirus (CMV) disease in transplant patients; the implications of clinical trials of prophylactic antiviral drug therapy; and unanswered questions about the optimal duration of prophylaxis and the risk of antiviral drug resistance are discussed. SUMMARY: CMV infection and disease are common in transplant patients. They often are the result of transplantation from a CMV-seropositive donor to a CMV-seronegative recipient or reactivation of latent virus in a CMV-seropositive recipient. Reactivation in transplant recipients may be the result of a blunted cytotoxic T-lymphocyte response and an imbalance in the release of pro-inflammatory and anti-inflammatory cytokines. CMV disease develops in many infected patients and frequently involves the graft. Antiviral drug therapies reduce the risk of and delay the onset of CMV disease in transplant recipients. They also reduce the risk of graft injury. CONCLUSION: The optimal duration of antiviral drug prophylaxis in transplant patients remains to be determined; however, it is clear that antiviral prophylaxis can prevent CMV disease as well as many of the complications associated with CMV disease. PMID- 15821266 TI - Epidemiology of cytomegalovirus disease in solid organ and hematopoietic stem cell transplant recipients. AB - PURPOSE: The novel and traditional risk factors for cytomegalovirus (CMV) infection and disease in solid organ transplant (SOT) and hematopoietic stem cell transplant (HSCT) recipients, the current strategies for the prevention of CMV disease, the emerging syndrome of late-onset CMV disease, and the risk factors for antiviral drug resistance are described. SUMMARY: The evolution of transplantation practices, the changes in the characteristics of donors and recipients, the introduction of novel immunosuppressive drugs, and the widespread use of antiviral drugs for CMV prevention have collectively influenced the natural history of CMV infection and disease in SOT and HSCT recipients. Strategies for CMV prevention with the use of antiviral drugs, whether as prophylaxis or as preemptive therapy, are effective in reducing the incidence of CMV disease in transplant recipients. However, late-onset CMV disease is now emerging as a serious problem. Donor and recipient CMV serostatus and other traditional risk factors for early-onset CMV disease appear to contribute to the occurrence of late-onset CMV disease during the first year after transplantation. CONCLUSION: The epidemiology of CMV infection and disease has changed after transplantation. Strategies to prevent late-onset CMV disease include the prolonged use of antiviral prophylaxis, but this practice can lead to the emergence of antiviral drug resistance, which has been associated with high rates of morbidity and mortality. PMID- 15821267 TI - Nature is vital to drug product discovery. PMID- 15821268 TI - Regions work to exchange health information. PMID- 15821270 TI - Adoption of duty-hour standards in a pharmacy residency program. PMID- 15821271 TI - Clinical leadership. PMID- 15821272 TI - Lesson from the saga of selective serotonin-reuptake inhibitors. PMID- 15821273 TI - Simplifying the treatment of HIV infection with ritonavir-boosted protease inhibitors in antiretroviral-experienced patients. AB - PURPOSE: The rationale, advantages, and disadvantages of attempting to enhance the efficacy of a primary protease inhibitor (PI) with ritonavir in the management of HIV infection, especially in patients who have previously undergone highly active antiretroviral therapy (HAART), are discussed. SUMMARY: PIs are pivotal components of the HAART regimens used to fight HIV infection. Long-term viral suppression remains a major clinical challenge. Certain pharmacologic features of many PIs, such as their limited oral bioavailability, necessitate burdensome dosage schedules, creating a barrier to patient adherence. Compliance may be further compromised by adverse events. Any factors that undermine adherence may increase the risk that plasma drug concentrations will be suboptimal and that viral resistance and subsequent treatment failure will develop. The pharmacokinetic enhancement, or "boosting," of PI levels with low dose ritonavir may increase PI potency and efficacy, as well as decrease the emergence of viral resistance, reduce the pill burden, and simplify administration. A number of clinical studies suggest that PI-boosted regimens are safe and effective in HIV-infected patients who have been previously treated with antiretroviral agents, but more research is needed. CONCLUSION: PI boosting with ritonavir can improve PI pharmacokinetics so that potency and efficacy are increased and regimens are simplified, thereby potentially reducing antiretroviral resistance and promoting patient adherence. PMID- 15821274 TI - Properties of pramlintide and insulin upon mixing. AB - PURPOSE: The pharmacokinetics, pharmacodynamics, and safety of pramlintide and various insulin formulations in patients with type 1 diabetes mellitus (DM) when given as separate injections or mixed in the same syringe before injection were studied. METHODS: In two randomized, open-label, placebo-controlled, five-period crossover studies, patients with type 1 DM received preprandial injections of pramlintide, short-acting insulin, and long-acting insulin administered either by separate injections or after mixing in various combinations. Serum free insulin and plasma glucose concentrations were measured for 10 hours and plasma pramlintide concentrations for 5 hours after injection. RESULTS: Blood samples were collected from a total of 51 patients. All treatments involving mixtures were comparable to separate injections with respect to the area under the concentration-versus-time curve (AUC) and the maximum concentration (Cmax) of serum free insulin. There were some minor differences in the AUC and Cmax of pramlintide. No injection-site reactions or other unexpected adverse events were observed. CONCLUSION: Mixing pramlintide with short- or long-acting insulin in the same syringe before subcutaneous injection did not affect the pharmacodynamics of glucose or the pharmacokinetics of insulin or pramlintide in a clinically significant manner. PMID- 15821275 TI - Prediction of gentamicin peak and trough concentrations from six extended interval dosing protocols for neonates. AB - PURPOSE: The ability of six extended-interval gentamicin dosing protocols to achieve desired peak and trough concentrations in neonates was evaluated using simulated calculations based on pooled patient data. METHODS: The demographic and pharmacokinetic data of 293 neonates age one week or less from three previously published studies were pooled and applied in each of six published protocols. The data collected, including weight, dosage, dosing interval, and measured serum or plasma gentamicin concentrations, were used to determine gentamicin clearance, elimination-rate constant, and distribution volume. Peak and trough gentamicin concentrations were calculated and compared with several therapeutic ranges that aim to achieve high peak concentrations for maximum efficacy and low trough concentrations for minimum toxicity. RESULTS: All protocols resulted in more than 95% of peak concentrations exceeding 5 mg/L. Peaks exceeded 10 mg/L in 26-42% of the simulations. Most of the protocols resulted in high trough concentrations (0.5-1.5 mg/L). Trough concentrations were below 1 mg/L in 51-83% of the simulations. CONCLUSION: Simulated calculations based on pooled patient data found that six gentamicin dosing protocols for neonates would likely yield acceptable peak concentrations and a trough concentration of < or = 1 mg/L for at least 50% of patients. It may be sufficient to monitor only the trough concentrations when applying these protocols. PMID- 15821276 TI - Compatibility and osmolality of inhaled N-acetylcysteine nebulizing solution with fenoterol and ipratropium. AB - PURPOSE: The compatibility, pH, and osmolality of N-acetylcysteine (NAC) nebulizing solution in the presence of ipratropium bromide or fenoterol hydrobromide were studied. METHODS: Portions (400 microL) of each mixture were sampled immediately upon mixing and one, two, three, four, five, six, and seven hours after mixing and assayed by high-performance liquid chromatography. Osmolality was measured by sampling 100 microL from the filling cup at a five minute interval during nebulization and by the freezing-point-depression method. RESULTS: Adding NAC solution to fenoterol solution raised the pH from 3.20 to 7.90 and the osmolality to a mean +/- S.D. of 1400.67 +/- 4.51 mOsm/kg. Fenoterol concentrations decreased to 93.71% and NAC concentrations to 92.54% of initial concentrations after seven hours. Mixing ipratropium with NAC solution raised the pH from 3.74 to 7.95 and the osmolality to a mean +/- S.D. of 1413 +/- 11.79 mOsm/kg. The initial ipratropium concentration declined 7.39% and 10.91% one and two hours after mixing with NAC solution, respectively. CONCLUSION: NAC and ipratropium were stable in nebulizing solution within one hour of mixing. NAC and fenoterol were compatible for at least seven hours. PMID- 15821277 TI - Compatibility of anidulafungin with other drugs during simulated Y-site administration. PMID- 15821278 TI - Reason and history as guides for hospital pharmacy practice leaders. PMID- 15821279 TI - Selective serotonin-reuptake inhibitors and suicidal ideation and behavior in children. PMID- 15821280 TI - Oxygen-dependent gene expression in fishes. AB - The role of oxygen in regulating patterns of gene expression in mammalian development, physiology, and pathology has received increasing attention, especially after the discovery of the hypoxia-inducible factor (HIF), a transcription factor that has been likened to a "master switch" in the transcriptional response of mammalian cells and tissues to low oxygen. At present, considerably less is known about the molecular responses of nonmammalian vertebrates and invertebrates to hypoxic exposure. Because many animals live in aquatic habitats that are variable in oxygen tension, it is relevant to study oxygen-dependent gene expression in these animals. The purpose of this review is to discuss hypoxia-induced gene expression in fishes from an evolutionary and ecological context. Recent studies have described homologs of HIF in fish and have begun to evaluate their function. A number of physiological processes are known to be altered by hypoxic exposure of fish, although the evidence linking them to HIF is less well developed. The diversity of fish presents many opportunities to evaluate if inter- and intraspecific variation in HIF structure and function correlate with hypoxia tolerance. Furthermore, as an aquatic group, fish offer the opportunity to examine the interactions between hypoxia and other stressors, including pollutants, common in aquatic environments. It is possible, if not likely, that results obtained by studying the molecular responses of fish to hypoxia will find parallels in the oxygen-dependent responses of mammals, including humans. Moreover, novel responses to hypoxia could be discovered through studies of this diverse and species-rich group. PMID- 15821281 TI - Regulator of G protein signaling (RGS2)-deficient mice: a novel model to study autonomic nervous system function. PMID- 15821282 TI - Vioxx, Celebrex, Bextra....do we have a new target for anti-inflammatory and antipyretic therapy? PMID- 15821283 TI - Expression of membrane-associated carbonic anhydrase isoforms IV, IX, XII, and XIV in the rabbit: induction of CA IV and IX during maturation. AB - Several carbonic anhydrase (CA) isoforms are associated with plasma membranes. It is probable that these enzymes interact with anion transporters to facilitate the movement of HCO3- into or out of the cell. A better knowledge of CA isoform expression in a given tissue would facilitate a systematic examination of any associations with such transporters. We examined the expression of CAs IV, IX, XII, and XIV mRNAs in rabbit tissues, including kidney, heart, lung, skeletal muscle, liver, pancreas, gall bladder, stomach, small intestine, colon, and spleen, using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). CA IV mRNA was mainly in kidney, heart, lung, colon, and gall bladder. CA IX mRNA was restricted to stomach, gall bladder, duodenum, and early jejunum. CA XII mRNA was found in kidney and colon. CA XIV mRNA was localized to heart, lung, skeletal muscle, and liver. The data indicate that there are different patterns of CA expression in various tissues: CA IX was expressed in the proximal gastrointestinal tract, whereas CA XII and CA IV were more distal. CA IV and CA XII are important kidney isoforms. CA XIV was abundant in metabolically active tissues such as liver, heart, lung, and skeletal muscle. Some significant species differences were noted in the expression of some of these isoforms; for example, CA XIV is not expressed in rabbit kidney, despite being abundant in mouse kidney. Maturational studies showed that the expression of CA IX mRNA and protein increased markedly with weaning ( approximately 3-4 postnatal wk) and was well correlated with the maturational expression of the alpha-subunit of the gastric H+,K+-ATPase, suggesting that function of CA IX and the gastric H+ pump might be linked in the digestion of adult foodstuffs. The unique pattern of membrane-bound CA isoforms suggests different functional associations with transporters, depending on the physiological demands on the tissue. PMID- 15821284 TI - Changes in PKB/Akt and calcineurin signaling during recovery in atrophied soleus muscle induced by unloading. AB - Protein kinase B [PKB, also known as Akt (PKB/Akt)] and calcineurin (CaN) are postulated to play important roles in integrating intracellular signaling in skeletal muscle in response to disuse and increased muscle loading. These experiments investigated changes in signal transduction of the downstream pathways of PKB/Akt and CaN during recovery following disuse-induced muscle atrophy. A 10-day period of hindlimb unloading (HLU) via tail suspension (male rats) was used to produce soleus muscle atrophy. Muscle recovery was achieved by returning animals to normal ambulation for 3-10 days. HLU resulted in significant muscle atrophy and a slow-to-fast fiber transition as revealed by appearance of type IId/x and IIb myosin heavy chain (MHC) isoforms. Muscle mass in HLU animals recovered to control (Con) levels after 10 days of reloading, but the fast-to slow shift in muscle MHC was incomplete, as indicated by the continued presence of type IId/x MHC. Ten days of HLU resulted in a significant decrease (-43%) in muscle levels of phosphorylated PKB/Akt. In contrast, muscle levels of phosphorylated PKB/Akt were greater (+56%) in HLU than in Con animals early after the onset of reloading (3 days). Soleus levels of phosphorylated p70S6K were significantly higher (+26%) in HLU animals after 3 days of muscle reloading. Muscle levels of phosphorylated PKB/Akt and phosphorylated p70S6K returned to Con levels by day 10 of recovery. Moreover, muscle CaN levels were significantly higher than Con levels after 10 days of muscle reloading. These findings are consistent with the hypothesis that PKB/Akt and its downstream mediators are active in the regrowth of muscle mass during the early periods of recovery from muscle atrophy. Our data support the concept that CaN is involved in muscle remodeling during the later phases of recovery from disuse muscle atrophy. PMID- 15821285 TI - Short photoperiod exposure increases adipocyte sensitivity to noradrenergic stimulation in Siberian hamsters. AB - Siberian hamsters (Phodopus sungorus) exhibit a naturally occurring, reversible seasonal obesity with body fat peaking in long "summerlike" days (LDs) and reaching a nadir in short "winterlike" days (SDs). These SD-induced decreases in adiposity are mediated largely via sympathetic nervous system (SNS) innervation of white adipose tissue (WAT), as indicated by increased WAT norepinephrine (NE) turnover. We examined whether SDs also increase sensitivity to NE-stimulated lipolysis. This was accomplished by measuring NE- and beta3-adrenoceptor (beta3 AR) agonist (BRL-37344)-induced lipolysis (glycerol release) as well as NE induced cAMP accumulation by inguinal, epididymal, and retroperitoneal WAT (IWAT, EWAT, and RWAT) in isolated adipocytes of LD- and SD-housed hamsters. SDs increased potency/efficacy of NE-triggered lipolysis in a temporally and fat pad specific manner. Thus when WAT pad mass decreased most rapidly (5 wk of SDs), potency (sensitivity/EC50) and efficacy (maximal response asymptote) of NE stimulated lipolysis were increased for all WAT pads and also at 10 wk for IWAT compared with their LD counterparts. SD enhancement of lipolysis was similar for NE and BRL-37344 in IWAT adipocytes. These results, coupled with our previous demonstration that SDs upregulate WAT beta3-AR mRNA expression, suggest that increased beta3-ARs mediated the SD-induced increased NE sensitivity. NE stimulated adipocyte accumulation of cAMP was greater after 5 wk of SDs for IWAT and EWAT and after 10 wk of SDs for IWAT compared with LDs, with no photoperiod effect for RWAT. Therefore, the SD-induced increase in SNS drive to WAT and increased sensitivity to this drive may work together to increase lipolysis in SDs. PMID- 15821286 TI - Taste discrimination between NaCl and KCl is disrupted by amiloride in inbred mice with amiloride-insensitive chorda tympani nerves. AB - The amiloride-sensitive salt transduction pathway is thought to be critical for the discrimination between sodium and nonsodium salts in rodents. In rats, lingual application of amiloride appears to render NaCl qualitatively indistinguishable from KCl. In this study, we tested four strains of mice for salt discriminability. In one strain (C57BL/6J), chorda tympani nerve (CT) responses to NaCl are attenuated by amiloride, and in the other three strains (BALB/cByJ, 129P3/J, DBA/2J) they are not. Under water-restriction conditions, these mice (7 mice/strain) were trained in a gustometer to lick for water from one reinforcement spout in response to a five-lick presentation of NaCl and to lick from another in response to KCl [salt concentration was varied (0.1-1 M) to render intensity irrelevant]. Mice were then tested with the stimuli dissolved in amiloride hydrochloride, and the latter was used as the reinforcer as well. Each concentration of amiloride (0.1-100 microM) was used on 2 separate days with control sessions interposed. Mice from all four strains were able to discriminate NaCl from KCl reliably. Amiloride impaired this discrimination in a dose dependent fashion. Moreover, performance on NaCl trials appeared to be more affected by amiloride than that on KCl trials in all four strains. Thus, in contrast to the predictions based on CT recordings, discrimination in all four strains appeared to depend on the amiloride-sensitive transduction pathway, which, in the case of BALB/cByJ, 129P3/J, and DBA/2J (and perhaps C57BL/6 as well), may exist in taste buds innervated by nerves other than the CT. PMID- 15821287 TI - Biochemical characterization of plasma membrane H+-ATPase activation in guard cell protoplasts of Arabidopsis thaliana in response to blue light. AB - Recent genetic analysis showed that phototropins (phot1 and phot2) function as blue light receptors in stomatal opening of Arabidopsis thaliana, but no biochemical evidence was provided for this. We prepared a large quantity of guard cell protoplasts from Arabidopsis. The immunological method indicated that phot1 was present in guard cell protoplasts from the wild-type plant and the phot2 mutant, that phot2 was present in those from the wild-type plant and the phot1 mutant, and that neither phot1 nor phot2 was present in those from the phot1 phot2 double mutant. However, the same amounts of plasma membrane H+-ATPase were found in all of these plants. H+ pumping was induced by blue light in isolated guard cell protoplasts from the wild type, from the single mutants of phototropins (phot1-5 and phot2-1), and from the zeaxanthin-less mutant (npq1-2), but not from the phot1 phot2 double mutant. Moreover, increased ATP hydrolysis and the binding of 14-3-3 protein to the H+-ATPase were found in response to blue light in guard cell protoplasts from the wild type, but not from the phot1 phot2 double mutant. These results indicate that phot1 and phot2 mediate blue light dependent activation of the plasma membrane H+-ATPase and illustrate that Arabidopsis guard cell protoplasts can be useful for biochemical analysis of stomatal functions. We determined isogenes of the plasma membrane H+-ATPase and found the expression of all isogenes of functional plasma membrane H+-ATPases (AHA1-11) in guard cell protoplasts. PMID- 15821288 TI - Fungal elicitor induces singlet oxygen generation, ethylene release and saponin synthesis in cultured cells of Panax ginseng C. A. Meyer. AB - Singlet oxygen is a high-energy molecular oxygen species. As one of the most active intermediates involved in chemical and biochemical reactions, singlet oxygen plays essential roles in plant responses to UV and strong light. Here, we report that Cle, an elicitor derived from fungal cell walls, induces the generation of singlet oxygen in cell cultures of ginseng, Panax ginseng. Cle treatment also triggers the activation of plasma membrane NADPH oxidase and 1 aminocyclopropane-1-carboxylic acid oxidase (ACO), subsequently leading to ethylene release and increased saponin synthesis, as shown by increased mRNA expression of squalene synthase (SQS) and squalene epoxidase (SQE), and accumulation of beta-amyrin synthase (beta-AS). Suppression of Cle-induced singlet oxygen generation or inhibition of ethylene production blocks saponin synthesis, whereas treatment of ginseng cells with ethylene or singlet oxygen induces the synthesis of saponin. Together, these results indicate that Cle induced production of both singlet oxygen and ethylene is required for saponin synthesis, and that singlet oxygen may function upstream of ethylene during Cle induced saponin synthesis. PMID- 15821293 TI - The expanding role of the pediatrician in improving child health in the 21st century. AB - Children in the United States increasingly are at risk for health problems that are precipitated or exacerbated by social, community, and environmental factors. Currently, pediatricians are unable to sufficiently address these health conditions without expanding their roles beyond that of providing health care to individual patients. Thus, to improve child health, physicians must work within their communities to identify the needs of the population they serve and take appropriate action to influence the private and public policies that address these needs. Healthy People 2010 establishes a well-supported framework that confronts the social and community factors that affect children's health and serves as a resource for community-minded pediatricians. In addition to Healthy People 2010, other successful initiatives have been created by pediatricians, and they must be expanded if the pediatric community is to alleviate the social, community, and environmental factors that negatively affect child health. PMID- 15821294 TI - Successes and missed opportunities in protecting our children's health: critical junctures in the history of children's health policy in the United States. AB - This article revisits several turning points in the history of child health policy for the purpose of understanding why many current health needs of children have not been addressed. We demonstrate how the rupture of ties between child medical and child welfare leaders, as well as the fault lines between various health care professionals, led to difficulties in establishing programs for children in the early 20th century. We note how wartime mobilizations helped to make the needs of the nation's youth apparent to political leaders and observe that programs begun in response to these discoveries often were ended in peacetime. Finally, we discuss how politics shaped the situation wherein maternal and child health programs, including Medicaid, are need based, severely underfunded, and administered by the states, whereas benefit programs for the elderly, including Medicare and Social Security, are general entitlements administered at the federal level. PMID- 15821295 TI - Commentary: Successes and missed opportunities. PMID- 15821296 TI - Community pediatrics: the Rochester story. AB - There are so many problems facing children today (eg, violence, poor nutrition, substance abuse, teen pregnancy) that conventional medical care can only address a small portion of these concerns. Thus, to be optimally effective, pediatrics needs to be linked to other disciplines and programs that address these issues by using different paradigms. Robert Haggerty, the originator of the term "community pediatrics," reflects on how one can successfully practice community pediatrics in an academic setting and model it for young physicians while also improving the health of children at the community level. Here we tell the story of the years that Haggerty was chief of pediatrics at the University of Rochester and took on the challenge of fulfilling the department's responsibility to all children in the county. Because of his pioneering work, his tenure was heralded as a critical period in the development of the field of community pediatrics. PMID- 15821297 TI - Community-level child health: a decade of progress. AB - In the late 1980s, the leadership of the South Carolina Chapter of the American Academy of Pediatrics (AAP) developed a growing awareness that there was a population of children in the state who frequented emergency departments and public health clinics but were not considered patients in any medical professional's office. Coincidentally, the commissioner of the South Carolina Department of Health and Environmental Control was struggling to expand resources needed to care for these children, who were crowding his facilities in search of primary care. Department of Health and Environmental Control clinic staff increasingly were unable to provide core public health services because of the need to treat illnesses and conduct well-child visits. Concurrently, with mounting concerns over the inadequacy and/or unavailability of quality health care for all children, the AAP embarked on the Access to Care campaign, a multiyear effort to improve access to care. The South Carolina Chapter of the AAP and Department of Health and Environmental Control subsequently launched an initiative, the Partnerships for Children program, to ensure that South Carolina children who were medically disenfranchised could have a medical home and all other services necessary for health and well-being. Although structured somewhat differently in various communities and practices, each of the >75 partnerships statewide focused on using public health professionals for specific care coordination and support that complemented physician office visits and encounters. These partnerships have gained substantial leverage through the expanding resources of the AAP Access to Care campaign. In reviewing claims for children <3 years old who were enrolled in Medicaid before the establishment of the Partnerships for Children program, <45% had evidence of a well-child visit. Data in the fourth year of the program indicate that 84% of children in this age group have had at least 1 well-child visit. Other data suggesting improved access to care are equally reassuring. In this report, I describe the evolution of the Partnerships for Children program, identify key ingredients for its success, and provide examples of effective strategies that bring community resources into pediatric practices to allow for more efficient and productive delivery of primary care. I also indicate the important role of the AAP Community Access to Child Health infrastructure in the maintenance and further development of this initiative. PMID- 15821298 TI - Evidence-based community pediatrics: building a bridge from bedside to neighborhood. AB - The American Academy of Pediatrics policy statement "The Pediatrician's Role in Community Pediatrics" encourages all pediatricians to partner with their communities to create and disseminate innovative programs that improve child health. This article describes 4 pillars of a bridge to evidence-based community pediatrics for pediatricians interested in pursuing effective community action: (1) collaborate with the community to establish a specific, short-term, health related goal; (2) identify evidence-based best practice(s) for achieving the shared goal; (3) collaborate with the community to adapt this best practice to the community's unique assets and constraints; and (4) evaluate the project by using appropriate expertise. Practical elements of each pillar are described and illustrated by specific examples from community-based efforts of pediatricians and are accompanied by specific resources to aid pediatricians in their future community health work. PMID- 15821299 TI - Commentary: Pre-primary care pediatrics. PMID- 15821300 TI - Teaching community pediatrics to pediatric residents: strategic approaches and successful models for education in community health and child advocacy. AB - To improve child health at a community level, pediatricians require knowledge and skills that have not been traditionally included in residency training. Recent policy statements from the American Academy of Pediatrics and requirements from Accreditation Council for Graduate Medical Education Residency Review committees emphasizing the importance of community pediatrics training have provided additional incentive for pediatric residency programs to actively explore methods of teaching the principles and promoting the practice of community pediatrics to resident trainees. With a growing number of diverse educational models in various stages of practice or development, common themes and approaches to promote successful teaching of community health and child advocacy can be described. This article defines strategies for 2 critical elements of community pediatrics training, engaging residents and building strong community partnerships, then highlights a number of educational models that illustrate key curricular components and methods. Published results from evaluations of some programs suggest that community pediatrics training of this caliber will cultivate a cadre of pediatricians (academic and community based, generalists and subspecialists, researchers and practitioners) who understand child health in the context of community and have the leadership and collaborative skills to improve the health of children in their communities. PMID- 15821301 TI - Commentary: Educational planning to achieve the goals of community pediatrics. PMID- 15821302 TI - Communities as teachers: learning to deliver culturally effective care in pediatrics. AB - A patient's culture has an effect on her or his view of illness, decision to seek care, and adherence to treatment plans and follow-up visits. In this article, we describe community-academic partnerships designed to teach improved delivery of culturally effective care conducted in pediatric residency training programs in New York, New York, and San Diego, California. Columbia University-Children's Hospital of New York-Presbyterian focuses most of residents' cultural-training experiences within 1 community program, a home-visitation program (Best Beginnings) with which residents work in various capacities throughout residency. The University of California, San Diego and Naval Medical Center San Diego use a series of cultural "immersion experiences" as a primary method. The creation of community-academic partnerships for the purpose of service and training can be a critical asset in the development of culturally effective care training: community partners become teachers and local communities serve as classrooms. PMID- 15821303 TI - Commentary: Cultural effectiveness--ask and listen. PMID- 15821304 TI - Achieving consensus on competency in community pediatrics. PMID- 15821305 TI - Competency in community pediatrics: consensus statement of the Dyson Initiative Curriculum Committee. PMID- 15821306 TI - Commentary: Achieving consensus on competency in community pediatrics. PMID- 15821307 TI - Building healthier communities for children and families: applying asset-based community development to community pediatrics. AB - Social capital is the power of social networks and relationships, which constitute the social environment. Social capital has been associated with many measures of health and development. Asset-based community development (ABCD) provides a framework to increase social capital and build stronger, healthier communities for children. ABCD is a strength-based approach to community building that emphasizes bringing together community assets including individual community members, voluntary associations, and institutions. How pediatricians can apply ABCD to child health is described. PMID- 15821308 TI - Perspectives on funding community pediatrics: a panel discussion of private and public funders. PMID- 15821309 TI - Commentary: Perspectives on funding community pediatrics. PMID- 15821310 TI - The essential role of research in community pediatrics. AB - Several recommendations in the American Academy of Pediatrics policy statement "The Pediatrician's Role in Community Pediatrics" underscore the essential role of research as an agent of change to promote the health and well-being of children. This article provides (1) a framework for thinking about research in community pediatrics, (2) special considerations important in conducting community-level research, (3) an example of community-level research that has significantly decreased mortality in children (prevention of sudden infant death syndrome), (4) an example of a current issue illustrating the importance of community pediatrics research (promotion of school readiness), and (5) a discussion of future directions for research. Many of the leading health problems facing the United States as outlined in Healthy People 2010 are problems that affect children or have their roots in childhood and are likely to be addressed by community-level research and interventions. It seems clear that pediatricians should be learning to participate in, advocate for, and conduct more community pediatrics research. PMID- 15821311 TI - Integrating pediatrics and public health. AB - The demographics of children in the United States and the determinants of child health outcomes are evolving rapidly. This evolution will require those responsible for children's health and well-being to acquire new skills and access new resources in the future to ensure the relevance of the field of pediatrics to the health of children. The objective of this article is to define a new paradigm for the practice of pediatrics that integrates the knowledge, attitudes, and skills of pediatrics with the principles, practice, and resources of public health to address the current challenges to child health in the United States. Historical and contemporary advances in the understanding of child health provide a precedent and framework to facilitate this integration. PMID- 15821312 TI - Commentary: Back to the future--reintegrating pediatrics and public health. PMID- 15821315 TI - [Through transhepatic drain during mechanical jaundice caused by secondary sclerosing cholangitis]. AB - 37 patients treated for mechanical jaundice caused by secondary sclerosing cholangitis were included in the study. In the study group (20 patients) biliary duct was drained by through transhepatic drainages, in control group (17 patients) this was done by Eahr and Vishnevsky method. Through transhepatic drain of biliary tree (postoperative complications were developed in 50% of cases, lethality-20%) performed during secondary sclerosing cholangitis, carrying out frame function, prevents further sclerosing of biliary duct and has appeared to be much more effective to remove the jaundice, cholangitis and liver insufficiency in comparison with the drain by Eahr and Vishnevsky (postoperative complications were developed in 70,6% of cases, lethality-41,2%). PMID- 15821316 TI - [Results of a surgical treatment of perforative gastroduodenal ulcers]. AB - The article gives an analysis of the surgical treatment of perforative gastroduodenal ulcers. Out of 185 patients in 143 (77,2%) cases took place operation on taking in a perforative hole when 9 (6,2%) patients, five of them above 60, couldn't get it over. It has to be noted that the mentioned 9 patients were taken to the hospital after 521 (five hundred and twenty one) hours the perforation had started, each of them after 58 (fifty eight) hours in average. At the same time for patients below 60 were taken to the hospital after 38 (thirty eight) hours, patients above 60 (sixty) after 74 (seventy four) hours. So one of the main causes of death was later hospitalization. This approach considerably reduces ulceroraphy due to the active implementation of the radical treatment methods. According to above given dates, the approach considerably reduces ulceroraphy due to the active implementation of some radical surgical treatment methods. PMID- 15821317 TI - [Sensitivity of PCR method for detection of mycobacteria tuberculosis in patients with lung tuberculosis]. AB - Both hospitalized patients and outpatient clinic patients at the National Centre of Tuberculosis and Lung Diseases of Georgia have been investigated. The group of patients with the tuberculosis of urogenital system, has been studied (newly detected cases), 70 cases in total. The examination of the urine was carried out by the Polymerase Chain Reaction (PCR) method in order to detect Kochi bacillus and by three-time bacterioscopy of urine on acid resistant bacterium. Mycobacterium tuberculosis in urine has been detected in 57 (81,43%) patients by PCR method, and by urine bacterioscopy acid fast bacilli (AFB)(+) in 36 (51,43%) patients. 50 hospitalized patients were examined as a separate group. They had the tuberculosis of lungs and insignificant pathological changes in urogenital system. Among them there was active bacillus secretion in 45 cases by phlegm bacterioscopy AFB(+). Out of 50 patients the mycobacterium tuberculosis in urine was detected in 30 (60%) cases by PCR method. It should be mentioned that according to the urine two-time bacterioscopy, carried out on 50 patients, has not been detected AFB(-) bacillus secretion. It may be concluded that the PCR method is advantageous in detecting of Kochi bacillus in urine. Introduction of this method in medical practice will give us the possibility to determine the risk-group for development of tubercular changes in urinary system associated with lung tuberculosis, in order to control such patients and to carry out the adequate urological examinations. PMID- 15821318 TI - [Usage of Cerasorbe in complex treatment of chronic generalized periodontitis (clinical-experimental study)]. AB - The effectiveness of the bioactive osteoplastic composition--Cerasorbe--at a surgical stage of combined treatment of chronic generalized periodontitis was examined. The results obtained in the study and the control groups prove, that Cerasorbe allows reduction of the treatment duration and achieves remission of the disease. From 127 patients with chronic generalized periodontitis 65 were operated by microplasmatic scalpel-irradiator as a part of a complex treatment. The treatment course comprised of 3-5 procedures. The quantity of manipulations was determined by the severity of the pathological process. Before and after the treatment the radiographical investigation was carried out in both groups. At the 18th month after the surgical intervention bone regeneration, confirmed by radiography, was shown in 87,8% cases in the study group and in 60,1% in the control group. The work presents experimental and morphological studies of regeneration of damaged areas of maxillo-facial bones. Time course of healing of defects in the lower jaw bone filled with Cerasorb was studied in experiment on rabbits. On days 7, 14, 21 and 28 four rabbits from each group were killed and the defects were investigation by X-ray and histological methods. We have stained the micropreparations by hematoxilin-eosine. In experiments with Cerasorb, bone regenerations replaced up to one half of the defect area by the day 28. We have shown high efficiency of Cerasorb in experiment and during complex treatment. Cerasorb stimulated reparative osteogenesis and can be recommended for using in the clinical practice. PMID- 15821319 TI - Prevention of coronary atherosclerosis by the use of combination therapy with antioxidant coenzyme Q10 and statins. AB - The goal of the present research was to assess the efficacy of combination treatment with antioxidant coenzyme Q10 and simvastatin as well as coenzyme Q10 without statin therapy in order to prevent coronary atherosclerosis. 42 outpatients were divided into 2 groups: receiving coenzyme Q10 (Hasco-Lek, Poland) 60mg daily and its combination with simvastatin (zocor, vasilip) 10mg daily for an 8-week period. The treatment with coenzyme Q10 demonstrated its potential independent role in positive modification of oxidative stress, antiatherogenic fraction of lipid profile, atherogenic ratio, platelet aggregability. Taking into consideration the obtained results the study supports the use of coenzyme Q10 in combination with statins. Suggested attractive approach may result in complete correction of dislipidemia, reverse of endothelial dysfunction, reduce degree of oxidative stress and platelet aggregability. Consequently such a combination may be beneficial in preventing of further development of atherosclerosis in native coronary arteries as well as in bypass grafts in all coronary heart disease patients with or without myocardial revascularization. PMID- 15821320 TI - [The problem of chronic refractory angina pectoris]. AB - Prognosis in the case of refractory angina pectoris is poor in comparison with stable angina. Patients with refractory angina are more likely to have an acute coronary attack as a result of more diffuse coronary artery lesions and a high prevalence of cardiac risk factors. The quality of life of these patients is also poor. 36 patients 35-70 years of age with chronic stable angina were studied (34 males and 2-females). In all patients the stable angina was diagnoses based on the results of coronary angiography, medical record, ECG, echocardiography, stress-test, blood glucose, coagulation and lipid spectrum tests results. All patients were divided into 2 groups. By coronary angiography in 5 cases severe stenosis (80-95%) and occlusion of left main coronary artery was founded. In 10 cases significant stenosis of main coronary artery was not found and the patients were referred for conventional anti-ischemic therapy. 11 patients were treated by revascularization procedures (4 coronary artery bypass grafting and 7 coronary angioplasty). 15 cases ( 41,6%) were considered as refractory angina pectoris as far as they were suffering from ischemic heart disease and symptoms were present despite optimal medical therapy. They had been considered not suitable for any interventional procedure due to several reasons (distal localization of stenosis, diffuse lesions of arteries, etc.). PMID- 15821321 TI - [Behavioral risk factors in patients with diabetes mellitus type II]. AB - Sixty patients with diabetes mellitus of the second type and thirty healthy persons were investigated. Structural interviews by Friedman, Rosenmann and Spilberger methods were conducted in all subjects. According to the results, out of four groups "0" group was related to the healthy subjects, who showed behavior of type "b". For the rest 1, 2, 3 groups, they reflected the pathological state of the "a" type behavior of various degrees. This type was documented in 90% of patients with diabetes mellitus. Severe "a" type behavior was mostly seen in males rather than in females that indicate to the males' deficiency in the development of the emotional sphere. These specific health aggravations are directly connected with the duration of the disease, its stage and emotional expressive receptivity of the "a" type. PMID- 15821322 TI - [Prevalence of minimal thyroid deficiency (subclinical hypothyreosis) and subclinical thireotoxicosis among females of average and elderly ages who hadn't been examined before]. AB - Adjara is a region in Georgia with high level at iodine deficiency. The research has been carried out to find subclinical dysfunctions of thyroid gland in women older than 40 years whose thyroid state hadn't been examined before. From 1998 to 2004 583 women aged 40-78 had been examined. Primary examination was done by different specialists. All women were tested on thyreotropic hormone (TSH) by means of immunodiagnostic unit "Digiscan SD-1000" (Austria). Examination of thyroid gland on the unit Philips SD-800 was done to estimate the volume and echological structure. The patients with raised level of TSH (more than 4,1) and lowered level of TSH (less 0,3) had undergone additional testing on free T4 and titers of antibodies to thyreoglobulin and thyreoperoxidase. All patients were divided according to their age. The 1st group n=289 from 40 to 55 (average age 44+/-1,3) and the 2nd group n=294 from 56 to 78 (average age 61,5+/-2,3). High frequency of subclinical hypothyreosis has been found in women from the first group 7,9 % as well as in the second group--17,7 %. Higher frequency of subclinical hypothyreosis was associated with the older age (p<0,001). In the first group the level of subclinical thyreotoxicosis was lower- 2,2 % and in the second group- 4,1 %. Higher frequency of subclinical hypothyreosis was associated with the older age as well. PMID- 15821323 TI - Variation of inspired nitric oxide and nitrogen dioxide concentrations during mechanical ventilation. AB - Inhaled Nitric oxide is a selective and potent pulmonary vasodilator with minimal influence on systemic vascular resistance. Due to these properties NO becomes widespread treatment for pulmonary hypertension of different etiologies. However, during inhalation via inspiratory limb of breathing circuit NO reacts with oxygen, producing nitrogen dioxide (NO2) which is known to be a toxic agent. Therefore, the exact measurements of NO and NO2 concentrations is very important during NO therapy, as was previously described by several authors. The aim of this study was to measure NO and NO2 concentrations at different sites of inspiratory limb of breathing circuit. Significant changes were found between different sites of circuit: Highest NO and lowest NO2 concentrations were detected at the distal points of circuit. We conclude, that NO delivery system should be connected with inspiratory limb of breathing circuit as distally as possible, to minimize contact of NO air with oxygen and to achieve maximal NO and minimal NO2 concentrations during inspiration. PMID- 15821325 TI - [Dynamics and structure of the neonatal mortality in Kutaisi and Imereti region]. AB - Dynamics of neonatal mortality and analysis of its structure in Kutaisi and in the region of Imereti during 1999-2000 are presented. Insufficiency in the organization of the prophylactic and treatment measures during the pregnancy and delivery and mistakes in the treatment, were determined according to the analyses of expert cards. Data of the State Department of Statistics and statistical counting of the labor houses and labor-helping setting of Kutaisi also were used. Analysis of the collected data shows high rates of perinatal and neonatal mortality in Kutaisi and birth reduction. However some trends in the reduction of these findings were occurred in the year 1996, which could be explained by the aids in the delivery blocks in the labor houses of Kutaisi. PMID- 15821324 TI - [Effectiveness of ronkoleukin in children with reactive arthritis of chlamydial etiology]. AB - 14 children with reactive arthritis (ReA) were treated by antibiotics of macrolid group--vilprafen and anti-inflammatory medicines. Immunocorrective mean ronkoleukin (recombinant interleukin-2) was used as an additional therapy for activation of lymphocytes and NK cells. Five patients from 14 were treated by intravenous injections of ronkoleukin followed by oral administration of this drug. 9 children were treated by oral administration of ronkoleukin. All 14 children were treated by ronkoleukin along with the basic therapy. All children got repeated course of ronkoleukin one year later. From 14 children 12 reached remission of the disease. PMID- 15821326 TI - [Effectiveness of treatment of bronchoobstruction in children with acute respiratory infections using home-made spacer]. AB - A metered-dose inhaler (MDI) with spacer is the best way to deliver bronchodilator therapy for treatment of bronchoobstructive syndrome, especially in children. In developing countries, commercially produced spacers are generally unavailable or are too costly. Hence, a home-made spacer modified from a 500 ml plastic bottle, was compared with imported spacer (AeroChamber) in 80 children with spastic bronchitis. Clinical score, PEF and oximetry were recorded at baseline and 15 min after treatment. A beta-2- agonist (Ventolin) was given via MDI and two spacers (home-made and conventional spacers). PEF and oxygen saturation were significantly increased at 15 min after administration of the ventolin in both groups. No significant difference was observed between Aerochamber and the home-made spacer. In conclusion, ventolin delivered by MDI with AeroChamber was as effective in bronchodilatation as MDI with home-made spacer. The home-made spacer therefore offers a simple, inexpensive and effective method for delivering aerosol drug. PMID- 15821327 TI - [Dependence of child health on the ecology]. AB - The purpose of the work was to establish correlation between detected with the help of micronuclei in exfoliation cells infringements in the genetic disorder of Acute Respiratory Infection (ARI) at children living in Lukhuni gorge. Heavy metals having mutagenous trait can cause variety of diseases and first of all influence on the genetic apparatus, which damage often reflects chromosome change. The study of micronuclei in exfoliation cells is comparatively new, easily reachable and believable informative method. From Lukhuni gorge it is indicated true lifting amount of micronuclei. In a distance this indications where far less. It is investigated 150 children: 50 from Lukhuni gorge (I group) where the maintenance of arsenic is sharply raised in environment and 100 from the regions remote from Lukhuni gorge. Amount average quantity micronuclei in 1000 cells in I group was very high 22,38+/-1,26 in. In the second was 4,36+/ 0,50 (t=12,82; p<0,0001). All children, under the anamnesis and the clinical data, have been divided on frequently were ill of Acute Respiratory Infection(in one year > 4-5 times) and seldom were ill of Acute Respiratory Infection (< 4-5 times). ARI most frequently were marked in this region. It is revealed correlation between amount of micronuclei in exfoliation cells and parameters of illness respiratory diseases at children living in territories, polluted with arsenic and his derivatives (R=0,789; p<0,000001). PMID- 15821328 TI - [Prospective analysis of demographic situation as a basic element for health system resources development]. AB - As in many Western industrialized nations, Georgians' fertility rate has fallen over the course of the 20th century from a relatively high level to a low one. In 1920, the average Georgian woman was expected to give birth to about 7,5 children in her lifetime; in 2003, that number had fallen to 1,4. This demographic transition is characteristic for industrial and industrializing nations and is usually associated with greater numbers of women joining the work force and increased divorce and cohabitation, all of which tend to reduce family size and drive down fertility rates. Similar patterns have emerged in the United States and other Western countries. This historic population decline has been met with increasing concern in some Georgian strata. The Georgian mass media have overflowed with alarming articles on population issues. Based on popular, nonprofessional interpretations of available vital statistics, some are calling "to save Georgia from depopulation." As a result, the general public has been misled about population issues. The average citizen is likely to draw a direct connection between the current economic slump and a demographic crisis. PMID- 15821329 TI - [Immunohistochemical characteristics of solitary bone cyst]. AB - In order to define the histogenesis of bone solitary cyst it was performed immunohistochemical study of surgical material obtained from 17 patients with the monoclonal antibodies against the following antigens: CD-31 (marker of endotheliocytes), IV type collagen (marker of blood vessel basal membrane), vimentin (marker of connective tissue) and Ki-67 (marker of mitotic cell). The results of the study showed, that solitary bone cyst shouldn't be attributed to the blood-vessel formations and because of its vimentin-positive peculiarity it has connective tissue genesis. At the same time bone cyst shouldn't be considered as tumoral process because it contains low quantity of mitotic cells' markers and the latter appears on the peripheral region of an active cyst, where the regeneration and intensive transformation of bone tissue take place. According to the above-mentioned the solitary cyst of bone should be considered as non-tumor connective-tissue pathology, histogenesis of which shouldn't be related to blood vessels. PMID- 15821330 TI - [The association of autoimmune thyroiditis with HLA-B8 in the Georgian population]. AB - The study of distribution of HLA antigens among Georgian population in case of autoimmune thyreoiditis revealed a statistically significant association of HLA B8 antigen with autoimmune thyreoiditis. For statistical analysis we used following criterion: relative risk, attributive risk, preventive fraction, correlation coefficient and immunogenetic correction (P). Without compatibility to these criteria, there were no associations of the HLA antigens with the diseases. Although 8 of the antigens gave relative risk more than one, but only HLA B8 gave real association with autoimmune thyreoiditis. Typing of HLA antigens of class I and class II loci was done by classic microlymphocytotoxic test. The test performed on the lymphocytes from patients with autoimmune thyreoiditis. The character of association of HLA antigens among Georgian population with Hashimoto disease differs in character from other populations that evidently can be explained by the distributional character of HLA antigens among normal Georgian population. PMID- 15821331 TI - The ESR study of redox state of hepatocytes during aging in white rats. AB - Aging is an inevitable biological process and characterized by a general decline in physiological function that leads to morbidity and mortality. Specific causes of this decline are not known. Rapid development in free radical biology and molecular technology has permitted the acquisition of data in support to the role of oxidative stress or injury as a major contributor to the aging process and to the pathogenesis of a number of diseases. Nevertheless, a direct link between oxidative stress and aging has not as yet been established. It is concluded that although there is considerable evidence implicating oxidative stress in the aging process, additional evidence is needed to clearly define the nature of the involvement. The present study has been designed to evaluate the age-related changes in rat liver by ESR method. The study was performed on 16 white male rats (4 newborns, 4 young, 4 adult and 4 senescent). Received data indicate that in white rats the aging is accompanied by slight increase in free radical production and significant decrease in antioxidant capacity of the liver cells. The present study suggests that accumulation of oxidized proteins during aging is most likely to be linked with an age-related decline of antioxidant enzyme activities. PMID- 15821332 TI - Background radiation of the territory of some military objectives in Georgia. AB - Background radiation on the territory of some military objects in Georgia has been investigated with the help of "DRPB-03" (dosimeter-radiometer) according to the specially created instruction. Background radiation of investigated objects varied from 0,09 to 0,28 mkZv/h; in average 0,18 mkZv/h. The maximal level was measured on the 8th object -0,28 mkZv/h, minimal on the 2nd object -0,09 mkZv/h, in average near the - 0,18 mkZv/h. The maximal level of the annual doze of radiation was measured on the 8th object -0,49 mZv/y; and minimal on the 2nd 0,15 mZv/y. The analysis of investigations showed, that the level of background radiation on the territory of the investigated military objects exceeds the level of European countries and South Caucasus (except Armenia). This could be explained by the peculiarities of our country: geomorphological structure and disposition of soil (altitude, latitude). These reasons could cause the high levels of natural background radiation. The level of background radiation is higher in Black Sea coast area, which may be the result of the Chernobyl Accident (contamination with artificial radionuclides). The comparatively high level of background radiation indicates that the personnel sustain relatively high radiation doses from natural radiation origins. PMID- 15821333 TI - [Clinical aspects of treatment of genital herpes with plaferon LB and phenowine]. AB - Exacerbation of genital herpes simplex is followed by suppression of immunocompetence of patients. This proves necessity of adjuvant immunocorrective therapy. In our previous investigations it was shown that plaferon LB possessed immunotropic effect, and phenowine (as an antioxidant remedy) was intensifying the action of plaferon. These data allow us to examine protective action of plaferon and phenowine in complex treatment of genital herpes. High-performance scheme of herpetic relapse was suggested, which was established on aetiotropic effect of acyclovir and immunomodulating action of plaferon and phenowine: satisfactory outcome in 92,1 percent (monotherapy with acyclovir--70,7%). Results of complex therapy appeared in reduction of period of acute infection (on average 2,3 days) and in prolongation of remission (on average 157,3 days, vs 88,5 days in control group). Therapeutic action was achieved by antiviral properties of acyclovir and plaferon, immunostimulating action of plaferon and antioxidant effect of phenowine. PMID- 15821334 TI - [Regulatory mechanisms of young men heart rate during psychoemotional load accompanied by irritation with different sounds]. AB - The goal of our research was to evaluate regulatory mechanisms of young men heart rate during psychoemotional load on the background of sound irritations of different characteristics. The study was a community trial and performed by single blind method on volunteer young men (age 18-22, n=73). As a method of description of heart rate regulation mechanisms, analysis of heart rate variability was selected. Psychoemotional load was studied by Landolt rings. We observed that only high frequency sound irritation increases mistakes and reaction time during psychoemotional load. We conclude that despite of initial status of organism regulation mechanisms, during high frequency sound irritation, quality of psychoemotional functions worsens. Increasing of sympathetic effect in persons with initial sympathetic domination in vegetative nervous system is observed. PMID- 15821335 TI - 32-nucleotide deletion, associated with defence against hiv/aids, is a predominant mutation of CCR5 gene in the population of Georgia. AB - There is a special interest to investigate genetic peculiarities in the populations with a low HIV seroprevalence. Despite of presence of high-risk conditions for rapid spread of HIV/AIDS epidemics in Georgia, the prevalence of this infection in the country remains very low. We studied polymorphisms of CCR5 gene in Georgians. Blood samples from 190 women randomly selected from the cohort of pregnant women involved in the program of prevention of mother-to-child HIV transmission in Georgia have been investigated. Two-step PCR was used to amplify the whole CCR5 genetic sequence. Detection of mutations and polymorphisms was done by dHPLC. All samples showing specific patterns by dHPLC, were sequenced to identify the exact nature of the mutation. It was shown that CCR5-delta32 mutation is a predominant alteration of CCR5 gene among Georgians. All subjects bearing this mutation were heterozygotes. Frequency of delta32 CCR5 allele in the population of Georgia was equal to 5%. Only one case of R223Q mutation and two cases of mutations in the non-coding region of CCR5 gene were also found. Our findings differ from the existing data showing the absence of the CCR5-delta32 mutation among Georgians and provide further support to the hypothesis on a Northeastern European origin of this mutation and North to South gradient of its distribution. PMID- 15821336 TI - Hepatoprotective effect of tectoridin and tectorigenin on tert-butyl hyperoxide induced liver injury. AB - To clarify the hepatoprotective effects of tectoridin and tectorigenin from Puerariae Flos, their effects on tert-butyl hyperoxide (t-BHP)-injured HepG2 cells and mice were investigated. When tectorigenin at a dose of 50 mg/kg was intraperitoneally administered to mice injured by t-BHP, it significantly inhibited the increase the activities of plasma ALT and AST by 39% and 41%, respectively, in the t-BHP-treated group. The inhibitory effect of tectorigenin is much more potent than that of a commercially available dimethyl diphenyl bicarboxylate. Orally administered tectoridin showed hepatoprotective activity. However, when tectoridin was intraperitoneally administrated to mice, no hepatoprotective activity was observed. Tectorigenin also protected against the cytotoxicity of HepG2 cells induced by t-BHP. This protection may have originated from the inhibition of apoptosis. Tectorigenin may be hepatoprotective and tectoridin should be a prodrug that is transformed to tectorigenin. PMID- 15821337 TI - Altered brain penetration of diclofenac and mefenamic acid, but not acetaminophen, in Shiga-like toxin II-treated mice. AB - It is well accepted that bacterial and virus infections elevate the levels of cytokines in serum and cerebrospinal fluids. Such high levels of cytokines might alter the integrity of the blood-brain barrier (BBB) and/or blood-cerebrospinal fluid barrier (BCSFB), subsequently affecting brain penetration of drugs. However, few reports have addressed this issue. Thus, we investigated brain penetration of cyclooxygenase (COX) inhibitors, commonly used as antipyretics, in mice treated with Shiga-like toxin II (SLT-II) derived from E. coli O157:H7, which significantly elevates cytokine levels. As antipyretics, we used diclofenac, mefenamic acid, and acetaminophen. We found that SLT-II significantly increased the brain-to-plasma concentration ratio (Kp) of diclofenac and mefenamic acid, but not of acetaminophen. Moreover, the Kp of diclofenac and mefenamic acid was increased by probenecid, an anionic compound. These results suggest that efflux anion transporters might be involved in the transport of diclofenac and mefenamic acid. Western blot analysis revealed that SLT-II decreased the expression of organic anion transporter-3, an efflux transporter located on the BBB and/or BCSFB. Taken together, these results suggest that SLT II and/or SLT-II-stimulated cytokines might change brain penetration of drugs and could possibly increase the risk of their side-effects by altering the expression of transporters. PMID- 15821338 TI - Naltrexone protects against hypotension, hyperthermia, and beta-endorphin overproduction during heatstroke in the rat. AB - Heat stroke is characterized by hyperthermia, arterial hypotension, decreased baroreflex sensitivity, and increased serum levels of beta-endorphin. Whereas naltrexone may have therapeutic potential in heat stroke, the underlying mechanism remains unclear. We tested the hypothesis that naltrexone may attenuate heat stroke by reducing hyperthermia, hypotension, decreased baroreceptor sensitivity, and/or increased serum levels of beta-endorphin. Heat stroke was induced by exposing the anesthetized adult Sprague-Dawley rats in an incubator at 43 degrees C. The moment in which the mean arterial pressure dropped irreversibly from the peak level was taken as the onset of heat stroke. Control rats were exposed to 24 degrees C. Mean arterial pressure, baroreceptor sensitivity, and maximal reflex bradycardia, after the onset of heat stroke, were all significantly lower than in control rats. However, rectal temperature and serum levels of beta-endorphin were all greater after the onset of heat stroke. Intravenous delivery of naltrexone (10 mg/kg) 20 min before the initiation of heat stress, but not immediately at the onset of heat stroke, significantly attenuated the above-mentioned reactions. Accordingly, naltrexone improved survival during heat stroke. These results suggest that naltrexone protects against hypotension and decrement of both baroreceptor sensitivity and maximal reflex bradycardia during heat stroke by reducing both hyperthermia and increment of serum beta-endorphin and thus improves survival. PMID- 15821339 TI - Characterization of forskolin-induced Ca2+ signals in rat olfactory receptor neurons. AB - Forskolin-induced Ca(2+) signals were examined in isolated rat olfactory receptor neurons (ORNs) using a Ca(2+) indicator, fura-2. In the soma of the ORNs, forskolin caused an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)) that was enhanced by a phosphodiesterase (PDE) 1 inhibitor, 8 methoxymethyl-3-isobutyl-1-methyl-xanthine, but not a PDE4 inhibitor, rolipram. Forskolin-induced Ca(2+) signals were abolished with the removal of extracellular Ca(2+) and un-affected by treatment with thapsigargin or caffeine plus ryanodine. Niflumic acid, a Ca(2+)-activated Cl(-) channel inhibitor, or nifedipine, an L type Ca(2+) channel inhibitor, slowed the initial rate of the increase in [Ca(2+)](i) in response to forskolin. Nifedipine did not affect the increase in [Ca(2+)](i) that was slowed by niflumic acid. In Ca(2+) measurements with a confocal microscope and a calcium indicator, Fluo-4, the onset of the response to forskolin in the knob region occurred simultaneously or earlier, but not later, than that in the soma. It is suggested that the forskolin-induced Ca(2+) signals are due to Ca(2+) influx, but not the release of Ca(2+) from Ca(2+) stores, and that the initial rapid increase in [Ca(2+)](i) is associated with the activation of the voltage-dependent Ca(2+) channels in rat ORNs. PMID- 15821340 TI - Involvement of adenosine A2a receptor in intraocular pressure decrease induced by 2-(1-octyn-1-yl)adenosine or 2-(6-cyano-1-hexyn-1-yl)adenosine. AB - The aim of the present study is to clarify the mechanism for the decrease in intraocular pressure by 2-alkynyladenosine derivatives in rabbits. The receptor binding analysis revealed that 2-(1-octyn-1-yl)adenosine (2-O-Ado) and 2-(6-cyano 1-hexyn-1-yl)adenosine (2-CN-Ado) selectively bound to the A(2a) receptor with a high affinity. Ocular hypotensive responses to 2-O-Ado and 2-CN-Ado were inhibited by the adenosine A(2a)-receptor antagonist 1,3,7-trimethyl-8-(3 chlorostyryl)xanthine (CSC), but not by the adenosine A(1)-receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX) or the adenosine A(2b)-receptor antagonist alloxazine. In addition, 2-O-Ado and 2-CN-Ado caused an increase in outflow facility, which was inhibited by CSC, but not by DPCPX or alloxazine. Moreover, 2-O-Ado and 2-CN-Ado increased cAMP in the aqueous humor, and the 2-O Ado-induced an increase in cAMP was inhibited by CSC. These results suggest that 2-O-Ado and 2-CN-Ado reduced intraocular pressure via an increase in outflow facility. The ocular hypotension may be mainly mediated through the activation of adenosine A(2a) receptor, although a possible involvement of adenosine A(1) receptor cannot be completely ruled out. 2-O-Ado and 2-CN-Ado are useful lead compounds for the treatment of glaucoma. PMID- 15821341 TI - Roles of SIRT1 and phosphoinositide 3-OH kinase/protein kinase C pathways in evodiamine-induced human melanoma A375-S2 cell death. AB - We previously demonstrated that evodimine isolated from Evodia rutaecarpa (Goshuyu in Japan) induced apoptosis in human malignant melanoma A375-S2 cells within 24 h. In this study, TUNEL assay also indicated that one cause of A375-S2 cell death induced by evodiamine was apoptosis. After treatment with evodiamine for the indicated time periods, anti-apoptotic protein SIRT1 expression was decreased; p53 expression and its phosphorylation were both enhanced, whereas transient induction of downstream p21 was not enough to promote cell cycle arrest. Inhibition of the phosphoinositide 3-OH kinase (PI3-K)/protein kinase C (PKC) survival pathway as well as subsequent inhibition of the ERK cascade might contribute to evodiamine-induced cell death. In addition, p53 activation in response to evodiamine administration was correlated with the activation of the PI3-K/PKC pro-apoptotic pathway, but did not require ERK participation. The inhibition of the PI3-K/PKC survival pathway might be responsible for SIRT1 inactivation and increased Bax/Bcl-2 expression ratio in evodiamine-induced cell death. PMID- 15821342 TI - Clinical and experimental evidence for prevention of acute renal failure induced by radiographic contrast media. AB - Acute renal failure still occurs as a complication after radiographic examination using iodinated radiocontrast medium. The incidence rate of radiocontrast medium induced nephropathy (radiocontrast nephropathy) is low (2 - 3%) in general. However, the rate is remarkably elevated in patients with pre-existing renal insufficiency. Radiocontrast nephropathy is associated with increased morbidity and mortality, particularly in patients with percutaneous coronary interventions. Although the reduction in renal blood flow and direct toxic action on renal tubular cells are considered to be involved, little is known about the etiology of radiocontrast nephropathy. A number of agents that improve renal circulation have been clinically tested for prevention of radiocontrast nephropathy, but none of them has succeeded. Protection of renal tubular cells against oxidative stress is another approach to avoid radiocontrast nephropathy. Prophylactic effects of antioxidants such as N-acetylcysteine and ascorbic acid have been reported by several investigators, although the effectiveness of these compounds is still a matter of debate. At present, hydration is regarded as the only effective, though incomplete, prophylactic regimen for radiocontrast nephropathy. Recently, we have shown that caspase-dependent apoptosis is an important factor in the pathogenesis of radiocontrast nephropathy and clarified cellular mechanisms underlying the radiocontrast media-induced apoptosis. This review summarizes clinical and experimental evidence for the etiology and prevention of radiocontrast nephropathy. PMID- 15821343 TI - Pathophysiological roles of amyloidogenic carboxy-terminal fragments of the beta amyloid precursor protein in Alzheimer's disease. AB - Several lines of evidence suggest that some of the neurotoxicity in Alzheimer's disease (AD) is attributed to proteolytic fragments of amyloid precursor protein (APP) and beta-amyloid (Abeta) may not be the sole active component involved in the pathogenesis of AD. The potential effects of other cleavage products of APP need to be explored. The CTFs, carboxy-terminal fragments of APP, have been found in AD patients' brain and reported to exhibit much higher neurotoxicity in a variety of preparations than Abeta. Furthermore CTFs are known to impair calcium homeostasis and learning and memory through blocking LTP, triggering a strong inflammatory reaction through MAPKs- and NF-kappaB-dependent astrocytosis and iNOS induction. Recently, it was reported that CTF translocated into the nucleus, binding with Fe65 and CP2, and in turn, affected transcription of genes including glycogen synthase kinase-3beta, which results in the induction of tau-rich neurofibrillary tangles and subsequently cell death. Spatial memory of transgenic (Tg) mice overexpressing CT100 was significantly impaired and CTFs were detected in the neurons as well as in plaques of the Tg mice and double Tg mice carrying CT100 and mutant tau. In this review, we summarize observations indicating that both CTF and Abeta may participate in the neuronal degeneration in the progress of AD by differential mechanisms. PMID- 15821344 TI - Two visual processing pathways are targeted by gonadotropin-releasing hormone in the retina. AB - In fish the terminal nerve is comprised of a group of cells with somata adjacent to the olfactory bulb and processes that extend both anteriorly to the olfactory mucosa and posteriorly to the telencephalon. In teleost fish an additional group of axons extends along the optic tract and delivers putative neuromodulators to the retina. One peptide - gonadotropin-releasing hormone (GnRH) - has been implicated as a prime candidate neuromodulator based on electrophysiological evidence that exogenous application influences neural activity. Here we describe the expression patterns of two GnRH receptor subtypes in the retina of a teleost fish, Astatotilapia (Haplochromis) burtoni. The type 1 GnRH receptor (GnRH-R1) was expressed in cells of the amacrine cell layer - where lateral inputs affect the flow of visual information from photoreceptors to the brain - and in a distribution and location pattern similar to dopaminergic interplexiform cells. Immunohistochemical labeling of GnRH fibers revealed varicosities along terminal nerve axons near the amacrine cell layer and near cells immunoreactive for tyrosine hydroxylase, a dopaminergic cell marker. This finding supports an existing model that the terminal nerve forms synapses with dopaminergic interplexiform cells. Surprisingly, the type 2 GnRH receptor (GnRH-R2) was abundantly expressed in ganglion cells, which lie along the direct pathway of visual information to the brain. These data suggest that GnRH from the TN could broadly influence processing of retinal signals both in lateral processing circuits through GnRH-R1 and in the vertical throughput pathway through GnRH-R2. PMID- 15821345 TI - Regional specialization in pyramidal cell structure in the visual cortex of the galago: an intracellular injection study of striate and extrastriate areas with comparative notes on new world and old world monkeys. AB - Recent studies have revealed marked differences in the basal dendritic structure of layer III pyramidal cells in the cerebral cortex of adult simian primates. In particular, there is a consistent trend for pyramidal cells of increasing complexity with anterior progression through occipitotemporal cortical visual areas. These differences in pyramidal cell structure, and their systematic nature, are believed to be important for specialized aspects of visual processing within, and between, cortical areas. However, it remains unknown whether this regional specialization in the pyramidal cell phenotype is unique to simians, is unique to primates in general or is widespread amongst mammalian species. In the present study we investigated pyramidal cell structure in the prosimian galago (Otolemur garnetti). We found, as in simians, that the basal dendritic arbors of pyramidal cells differed between cortical areas. More specifically, pyramidal cells became progressively more spinous through the primary (V1), second (V2), dorsolateral (DL) and inferotemporal (IT) visual areas. Moreover, pyramidal neurons in V1 of the galago are remarkably similar to those in other primate species, in spite of large differences in the sizes of this area. In contrast, pyramidal cells in inferotemporal cortex are quite variable among primate species. These data suggest that regional specialization in pyramidal cell phenotype was a likely feature of cortex in a common ancestor of simian and prosimian primates, but the degree of specialization varies between species. PMID- 15821346 TI - Somatotopic organization of perioral musculature innervation within the pig facial motor nucleus. AB - The orbicularis oris and buccinator muscles of mammals form an important subset of the facial musculature, the perioral muscles. In many taxa, these muscles form a robust muscular hydrostat capable of highly manipulative fine motor movements, likely accompanied by a specialized pattern of innervation. We conducted a retrograde nerve-tracing study of cranial nerve (CN) VII in pigs (Sus scrofa) to: (1) map the motor neuron pool distributions of the superior and inferior orbicularis oris, and the buccinator, to test the hypothesis that perioral muscle motor neuron pools exhibit a somatotopic organization within the facial motor nucleus; and (2) test the hypothesis that portions of the superior orbicularis oris (SOO) motor neuron pool also exhibit a somatotopic organization, reflecting a potential compartmentalization of function of the rostral, middle, and caudal segments of this muscle. Cresyl violet histological staining showed that the pig facial motor nucleus was comprised of 7 well-defined subnuclei. Neuroanatomical tracers injected into these perioral muscles transported to the motor neuron pools of the lateral 4 of the 7 subnuclei of the facial motor nucleus. The motor neuron pools of the perioral muscles were generally segregated from motoneurons innervating other facial muscles of the rostrum. However, motor neuron pools were not confined to single nuclei but instead spanned across 3-4 subnuclei. Perioral muscle motor neuron pools overlapped but were organized somatotopically. Motor neuron pools of portions of the SOO overlapped greatly with each other but exhibited a crude somatotopy within the SOO motor neuron pool. The large and somatotopically organized SOO motor neuron pool in pigs suggests that the upper lip might be more richly innervated than the other perioral muscles and functionally divided. PMID- 15821347 TI - Interspecific variation in photo-induced GnRH plasticity among nomadic cardueline finches. AB - Changes in response to photoperiod are of fundamental importance to seasonal cycles in avian reproductive development. In this study we present data on photo induced changes in gonadotropin-releasing hormone (GnRH) immunoreactivity associated with the development of photorefractoriness in males of 3 closely related species of cardueline finch: common redpoll (Carduelis flammea), pine siskin (C. pinus) and white-winged crossbill (Loxia leucoptera). All are nomadic, mid- to high-latitude breeders with varying levels of reproductive flexibility: redpolls are typically seasonal, pine siskins more flexibly seasonal and white winged crossbills temporally opportunistic. Males were exposed to either long (20L:4D) or short days (5L:19D), beginning in January, and GnRH was assessed via immunocytochemistry 4.5 months later. Plasma LH, testis size, and molt score were also measured. Significant reductions in the number of irGnRH cells detected, cross-sectional cell area and optical density of these cells occurred in both redpolls and siskins on long, as compared with short, days. These decreases in irGnRH, coupled with spontaneous testicular regression and onset of molt were consistent with the development of absolute photorefractoriness on constant long days. In contrast, the tendency for numbers of GnRH immunopositive cells and mean measures of GnRH immunoreactivity (cell area, optical density and axonal fiber density) to decrease were much more moderate in white-winged crossbills held on long versus short days. Although none of the four measures of GnRH immunoreactivity differed significantly between treatments in crossbills, significant shifts in the frequency distribution of irGnRH neurons to include greater numbers of less immunoreactive cells suggested that antigenic GnRH content was reduced in many cells. These photo-induced shifts in optical densities of GnRH cells in long-day, as compared with short-day crossbills followed, to a lesser extent, the pattern in pine siskins and common redpolls. For all three species, patterns of decrease in GnRH immunoreactivity, gonadal regression and molt suggest that some form of refractoriness to photostimulation develops on constant long days. However, more moderate tendencies for GnRH immunoreactivity to decrease in white-winged crossbills, coupled with limited gonadal regression and molt on long days suggest that absolute photorefractoriness might develop more slowly in this taxon than in others, or alternatively, a condition arises that is more similar to relative photorefractoriness. These data indicate that relatively small differences in the response of the neuroendocrine system to stimulatory cues such as day length may lead to remarkably different capabilities for reproductive flexibility even within a closely related family of birds. PMID- 15821348 TI - Brain allometry in bumblebee and honey bee workers. AB - Within a particular animal taxon, larger bodied species generally have larger brains. Increased brain size usually correlates with increased behavioral repertoires and often with superior cognitive abilities. Bumblebees are eusocial insects that show pronounced size polymorphism among workers, whereas in honey bees size variation is much less pronounced. Recent studies suggest that within a given colony, large bumblebee workers are more efficient foragers and are better learners than their smaller sisters. Here we examine the allometric relationship between brain and body size of worker bumblebees and honey bees. We find that larger bees have larger brains and that most brain components show a similar size increase as the overall brain. One particular brain structure, the central body, is relatively smaller in large bumblebees compared to small bees. The same is true for the mushroom body lobes, whereas the mushroom body calyces, which receive sensory input, are not reduced in larger bumblebees or honey bees. Honey bees have relatively smaller brains, as well as smaller mushroom bodies, than bumblebee workers. We discuss why brain or mushroom body size does not necessarily correlate with the degree of a species' social organization. PMID- 15821349 TI - The inhibition of CA2+ influx induced by hypericin in cultured human retinal pigment epithelial cells analyzed by confocal imaging. AB - PURPOSE: Hypericin, a specific inhibitor of protein kinase C, has been reported to have potential as a therapeutic drug for proliferative vitreoretinopathy (PVR) in vitro and in vivo. In the present studies, we analyzed the dynamic changes in Ca2+ influx and free intracellular Ca2+ concentration ([Ca2+]i) of cultured human retinal pigment epithelial (RPE) cells after stimulation with hypericin in an attempt to elucidate its mechanism as a therapeutic drug for PVR. METHODS: RPE cells were plated in a special plastic dish and then stimulated with 100 nM phorbol-12-myristate-13-acetate (PMA) and/or 6 hypericin concentrations (0.5, 1, 2, 3, 4 and 5 microM), after which Ca2+ influx and [Ca2+]i were determined using the fluorescence Ca2+ dye fluo-3 AM and laser scanning confocal microscopy. RESULTS: The fluorescence in resting RPE cells was strong and distributed throughout the cells. The nucleus appeared more fluorescent than the cytoplasm. After stimulation with 0.5 microM hypericin, no obvious change of Ca2+ influx and [Ca2+]i was observed. In contrast, stimulation with higher concentrations of hypericin (1-5 microM) led to a rapid decrease in Ca2+ influx and [Ca2+]i, which was significantly different from those detected without hypericin (control experiments). In addition, no significant differences in [Ca2+]i were found between 1 and 5 microM hypericin used. Stimulation with hypericin, which was applied immediately after preincubation with PMA for 24 h did not further change Ca2+ influx and [Ca2+]i. CONCLUSION: In RPE cells, high concentrations of hypericin (1-5 microM) significantly inhibit Ca2+ influx and induce a decrease in [Ca2+]i. Therefore, hypericin has potential as a therapeutic drug for PVR maybe through its inhibition of the Ca2+ influx pathway. PMID- 15821351 TI - Tissue microarray analysis of C-MYC oncogene copy number changes in larynx carcinoma. AB - PURPOSE OF THE STUDY: Copy number changes and structural rearrangements of 8q in tumors from the head and neck region have been reported by numerous studies. To give better insights into the role of 8q abnormalities in larynx carcinoma, C-MYC copy number changes were studied in a large number of tumor materials. PROCEDURES: Fluorescent in situ hybridization with DNA locus-specific probe for C MYC genewas applied on tissue microarrays presented by 863 larynx tumors. RESULTS: 32.02% had increased copy number of C-MYC. Of these, 6.16% hadamplification and 25.86% had C-MYC gains. The analysis revealed a significant difference in the frequency of gains in primary and advanced tumors. Furthermore, we observed an association of gains with advanced clinical stage (I-III to IV). No association was found between the frequency of oncogene amplification and the tumor phenotype. CONCLUSIONS: These results suggest that the C-MYC gains are significant for the larynx tumor progression. PMID- 15821350 TI - Glycobiology of head and neck squamous epithelia and carcinomas. AB - An impressive variety of regulatory processes including cell adhesion and migration, proliferation, apoptosis and differentiation folding and routing of glycoproteins have been found to be mediated by specific lectin-carbohydrate interactions. This article summarizes the data on glycobiological aspects of differentiation of squamous epithelia in the head and neck region under physiological conditions and in cancer. The possible function of lectins in tumor development and invasiveness is debated. Introduction of labeled endogenous lectins as a tool for the study of functional glycomics at the cellular level in head and neck squamous epithelia and carcinomas enables a complex interpretation of studied data because these lectins are normally occurring in these tissues. The lectinology of Langerhans cells in head and neck squamous epithelia and carcinoma is also mentioned. Finally, the use of the described data in the diagnosis and prospectively in the treatment of head and neck squamous cell carcinoma is shown. PMID- 15821352 TI - Increased PGE2 levels in nonmalignant mucosa adjacent to squamous cell carcinoma of the head and neck. AB - OBJECTIVE: The cyclooxygenases (COX) 1 and 2 are the rate-limiting enzymes of prostaglandin E(2) (PGE(2)) synthesis, and the upregulation of COX-2 has been reported in tumors of different origins. The aim of our study was to quantify the PGE(2) expression in squamous cell carcinoma and surrounding mucosa, to analyze the potential of acetylsalicylic acid (ASA) for reducing PGE(2) levels in these tissues, and to improve our understanding of potential tumor-derived stimulation of surrounding mucosa by PGE(2). STUDY DESIGN AND SETTING: Intracellular PGE(2) levels in primary head and neck squamous cell carcinoma (HNSCC) and the surrounding mucosa at 1 and 2 cm distance were analyzed ex vivo by ELISA. Subsequently, we treated in vitro tumor and normal mucosal cells from turbinates with recombinant PGE(2) and ASA, and quantified intracellular PGE(2) levels. RESULTS: We observed high PGE(2) levels in the tumor samples and in tumor surrounding mucosa. The addition of PGE(2) and arachidonic acid to tumor cell cultures resulted in no further increase in intracellular PGE(2) levels, while ASA reduced PGE(2) levels by up to 40%. In normal epithelial cell cultures, less PGE(2) (6% of that found in the tumor cells) was expressed, but stimulation with PGE(2) resulted in levels comparable to those of the tumor samples. CONCLUSION AND SIGNIFICANCE: We conclude that HNSCC and the surrounding mucosa express high levels of PGE(2). This expression is reduced efficiently by ASA. We propose a stimulation of PGE(2) expression in the epithelium surrounding HNSCC by tumor derived PGE(2) with a potential impact on tumor growth. PMID- 15821353 TI - Inflammatory myofibroblastic tumor of the larynx in a 2-year-old male. AB - Inflammatory myofibroblastic tumor (IMT) is an uncommon neoplasm that is usually located in the lung in the pediatric population. These tumors contain a variety of cell types with the myofibroblast being dominant. When located in the upper airway, IMTs tend to be less aggressive, but have the potential for local invasion and recurrence. We present an unusual case of IMT in the pediatric larynx and review the medical literature describing the common locations, diagnosis, etiology, histology, and treatment of this tumor. The mainstay of treatment is complete surgical excision. Careful and frequent follow-up including frequent fiberoptic laryngoscopy and CT scans are recommended to evaluate for recurrence. More aggressive resection may be necessary if multiple recurrences occur. PMID- 15821354 TI - Temporal changes in postural sway caused by ultrashort-acting hypnotics: triazolam and zolpidem. AB - Two ultrashort-acting hypnotics, triazolam 0.25 mg and zolpidem 10 mg, were studied for their effects on equilibrium function in humans. Eight healthy male subjects participated in a double-blind, placebo-controlled study after informed consent. They subjected to static equilibrium tests, oculomotor tests and an assay of drug concentrations in the blood. Zolpidem was statistically significant in postural sway in tandem stance test, as defined by parametric values of tracing sum length and polygonal area of foot pressure center measured by a gait analysis system. In the tandem stance test, triazolam was statistically significant in postural sway only as defined by the polygonal area. However, in the Romberg test, the only statistically significant difference in zolpidem use was observed in polygonal area values. Blood concentrations of triazolam and zolpidem were found to closely correlate with the extent of postural sway in both tandem stance and Romberg tests. In this study, zolpidem with minimal muscle relaxant effect incurred imbalance more extensively than triazolam, which is known for its effect of muscle relaxation. In addition, gaze deviation nystagmus was observed only in zolpidem use in 5 of 8 subjects (62.5%). From these results, it is suggested that in the use of hypnotics, sway derives from the suppression of the central nervous system relevant to awakening rather than from muscle relaxation. The prior reference to blood concentrations of hypnotics should help improve safety care in minimizing loss of balance control and possible fall. PMID- 15821355 TI - The slit ventricle syndrome: advances based on technology and understanding. AB - Despite many articles on slit ventricle syndrome (SVS) over the last 25 years, accepted terminology regarding the definition of this condition is lacking. Any shunted individual with a severe headache disorder in the context of ventricles that are normal or smaller than normal can be said to suffer from SVS, even though there are at least five forms of the condition. Logical management of SVS requires an understanding of the specific pathogenesis of the problem in individual patients, whether based on monitoring of intracranial pressure (ICP) or observation at the time of shunt failure or symptoms. Overdrainage syndromes, whether intermittent proximal obstruction or low pressure states, are best managed with valve upgrades and the addition of devices that retard siphoning. Increased ICP without ventriculomegaly at the time of shunt failure is best managed by shunting devices that access the cortical subarachnoid space such as lumboperitoneal shunts or shunts involving the cisterna magna. Cranial expansion operations and subtemporal decompression should be limited to patients with craniofacial syndromes. PMID- 15821356 TI - The problematic slit ventricle syndrome. A review of the literature and proposed algorithm for treatment. AB - This review discusses the clinical difficulties with the so-called slit ventricle syndrome. It discusses why this terminology makes it difficult to formulate treatment algorithms. Slit ventricle syndrome should probably be known as the non compliant ventricle syndrome to avoid confusion of radiological 'slit ventricles' with the syndrome. Therapeutic measures should ideally focus on the underlying pathology related to the syndrome. For those patients with iatrogenic small calvaria and synostosis, subtemporal decompression would appear a low-risk and quick procedure. Calvarial expansion would be a second-line treatment. For those patients with a normal-sized calvarium and non-compliant ventricle syndrome, valve upgrade, adding an antisiphon device or programmable valve would appear a good first step. For patients in whom the diagnosis is unclear, a potential management plan is discussed. PMID- 15821357 TI - Lumbar CSF shunting preferentially drains the cerebral subarachnoid over the ventricular spaces: implications for the treatment of slit ventricle syndrome. AB - Based on a proposed pathophysiology of slit ventricle syndrome (SVS), we have hypothesized that lumboperitoneal shunting exerts effects in SVS patients by increasing the buffering capacity for raised intracranial pressure (ICP) via an increase in cerebrospinal fluid drainage from the cerebral subarachnoid space (SAS). We describe 3 SVS patients with patent lumbar subarachnoid drainage but under-functioning ventriculoperitoneal shunts (VPS) who presented with ventriculomegaly (not SVS), and persistence of shunt malfunction like symptoms. Revision of the VPS resulted in complete resolution of symptoms despite a finding of low pressure in the ventricular space. This supports the hypothesis that lumboperitoneal shunting preferentially drains the SAS over the intraventricular space and in these cases allows the 'SVS' ventricles to enlarge by creating a pressure gradient from ventricles to SAS through the cortical mantle. PMID- 15821358 TI - Choroid plexus and aquaporin-1: a novel explanation of cerebrospinal fluid production. AB - Aquaporins are selective water channel proteins that play a central role in the homeostasis of human body water. The choroid plexus (CP) is considered to be the main cerebrospinal fluid (CSF)-producing structure. In this study, six specimens of normal human CP obtained during surgery were analyzed by immunohistochemistry techniques for aquaporin-1 (AQP1) expression and distribution. Intense, uniformly distributed AQP1 immunostaining was observable in the apical but not the basolateral side of cuboid cells of the CP. Moreover, this polarized expression of AQP1 was weakly detectable in the endothelial cells of choroid microvessels and, with a different pattern, in the cells lining the tubules shaped into crypts. Selective AQP1 expression on the surface of the normal human CP might explain the role of CSF production by this complex structure. PMID- 15821359 TI - Spinal neurenteric cyst. Report of 4 cases and review of the literature. AB - Neurenteric cysts are rare congenital lesions of the spine that are lined with entodermal epithelium. We describe 4 unusual cases of neurenteric cysts in patients aged between 3 and 16 years. The cyst was situated anteriorly in 3 patients and posteriorly in 1. All the patients had varied clinical and radiological features. The child with dorsal neurenteric cyst had stigmata of spinal dysraphism along with other vertebral anomalies. Another child had an associated syringomyelia along with evidence of severe arachnoiditis. All 4 patients underwent laminectomy and total microsurgical excision of the cysts through a posterior approach. The patient with diastematomyelia developed early recurrence of the cyst and underwent reoperation. Three patients showed an excellent recovery after surgery. The child with arachnoiditis failed to recover neurologically. PMID- 15821360 TI - Spinal rhabdomyosarcoma in a child with lipomyelomeningocele. AB - Malignant tumors arising within dysrhaphic malformations are very rare and are mostly teratomas; so far, only one rhabdomyosarcoma has been reported in this context. We report another case of a girl with lipomyelomeningocele who developed a lumbar rhabdomyosarcoma 2 years after birth and primary closure of the neural tube defect. We present clinical, radiological and pathological findings, discuss possible mechanisms of malignant transformation and review the literature. PMID- 15821361 TI - Congenital peripheral facial palsy associated with cerebellopontine angle arachnoid cyst. AB - OBJECTS: A rare case of cerebellopontine angle arachnoid cyst leading to congenital peripheral facial palsy was presented. CLINICAL PRESENTATION: A 1-year old girl presented with peripheral facial paralysis since birth. Computed tomography and magnetic resonance imaging revealed left cerebellopontine angle arachnoid cyst causing moderate displacement of the brain stem. INTERVENTION: Retrosigmoid suboccipital craniotomy was performed and microsurgical resection of the cyst wall and fenestration of the cyst to the basal cisterns were achieved. CONCLUSIONS: Cerebellopontine angle arachnoid cyst should be considered as a potential cause of congenital peripheral facial palsy. PMID- 15821362 TI - Synchronous optic and pineal pilocytic astrocytomas in a paediatric patient with neurofibromatosis type 1. AB - A 12-year-old girl with neurofibromatosis type 1 presented with headache, visual acuity and visual field disturbance. Computed tomography and magnetic resonance imaging revealed an enhanced solid mass involving her right optic nerve and optic chiasm, and a cystic lesion in the pineal region that had resulted in obstructive hydrocephalus. An open biopsy of the right optic nerve tumour was performed, and it was histologically identified as a pilocytic astrocytoma. Local irradiation of 50 Gy to the optic pathway tumour was performed, and the tumour has remained stable for more than 29 months. On the contrary, the pineal cystic mass that was also histologically identified as a pilocytic astrocytoma showed marked enlargement within 5 months after a subtotal resection. Chemotherapy with cisplatin and vincristine was performed after a second surgery, and the pineal tumour has not re-grown in 18 months. To our knowledge, this is the first case report to describe synchronous optic and pineal pilocytic astrocytomas associated with neurofibromatosis type 1. PMID- 15821363 TI - Germinoma of the basal ganglia. An 8-year asymptomatic history after detection of abnormality on CT. AB - We describe a case of germinoma of the left basal ganglia. An 11-year-old boy, who demonstrated calcification of the left basal ganglia on CT scan following a head injury at the age of 3 years, presented with a weakness of the right upper extremity for 2 months. MRI demonstrated high intensity in the left basal ganglia on a T1-weighted image without enhancement as well as high intensity on a T2 weighted image. Ipsilateral hemiatrophy of the hemisphere and midbrain was also noted. In addition, high intensity in the left internal capsule and cerebral peduncle was demonstrated on T2-weighted image. Surgical specimens obtained by stereotactic biopsy showed germinoma with a two-cell pattern. The patient had remained asymptomatic for 8 years after abnormal calcification was initially detected on CT scan. Ipsilateral hemiatrophy of the hemisphere and midbrain was demonstrated before the onset of weakness. PMID- 15821364 TI - Spontaneous pneumocephalus presenting with apnea attacks in a newborn with open myelomeningocele. AB - We report a case who had an open myelomeningocele and developed respiratory arrest associated with spontaneous pneumocephalus. Spontaneous respiration resumed after closing of the defect and ventriculoperitoneal shunting. We conclude that repair of the defect in the early period is very useful both to minimize the infection risk and to prevent the complication of pneumocephalus in these cases. PMID- 15821365 TI - Lateral ventricle choroid plexus papilloma extending into the third ventricle. PMID- 15821366 TI - Accuracy of customized miniature stereotactic platforms. AB - In this study, a new system was evaluated for implanting deep-brain stimulators based on a one-piece platform for each trajectory customized from a preoperative planning image. During surgery, the platform is attached to skull-implanted posts that extend through the scalp. The platform acts as a miniature stereotactic frame to provide guidance for parallel cannulas as they are advanced through a burr hole to the target. Accuracy is determined from a postoperative CT. For each implantation, the distance between the position observed in the postoperative image and the position calculated relative to the platform from the preoperative image is our measure of error. Because this measure incorporates the surgical error of electrode anchoring, brain shift between preoperative and postoperative scanning, and error in the measurement of the position of the electrode in CT, it will tend to overestimate the true error. The mean error was 2.8 mm for 20 implantations. These data reflect favorably the accuracy of this system when compared with others. PMID- 15821367 TI - Venous air embolism during deep brain stimulation surgery in an awake supine patient. AB - Deep brain stimulation (DBS) of the subthalamic nucleus and globus pallidus is used to improve Parkinsonian symptoms and reduce levodopa-induced motor complications in Parkinson's disease (PD). This procedure is usually performed with minimal or no sedation to allow accurate feedback from patients during surgery. Venous air embolism (VAE) has been previously reported in patients undergoing awake neurosurgical procedures for brain tumors or pallidotomy for PD. We describe a case of intraoperative VAE in an awake, supine patient while undergoing DBS surgery for PD who presented with coughing, tachypnea and hypoxemia. The difference in clinical presentation between VAE in awake vs. anesthetized patients is discussed as are intraoperative monitoring techniques and management options. PMID- 15821369 TI - Health research in medical colleges. PMID- 15821368 TI - The application of optical recording of intrinsic signals to simultaneously acquire functional, pathological and localizing information and its potential role in neurosurgery. AB - INTRODUCTION: The accurate intraoperative localization of epileptic foci and surrounding functional architecture is critical to a successful surgical outcome. Current techniques are limited either by their inability to simultaneously sample large areas of cortex with high spatial resolution or account for dynamic alterations in cortical morphology. Optical recording of intrinsic signals can map neuronal activity in a large area of cortex with a spatial resolution in the order of <100 mum. We explored methods of simultaneously representing localizing information, functional architecture and the border of an epileptic focus in vivo with intrinsic signal imaging. METHODS: The functional architecture of V1 was mapped using optical imaging of intrinsic signals in the ferret at 707 nm (n = 9). Interictal and ictal foci were then generated with focal iontophoresis of bicuculline methiodide and 4-aminopyridine into V1 and mapped optically. Blood vessel architecture was mapped using light acquired at 540 nm. RESULTS: Epilepsy maps could be superimposed on maps of the underlying functional architecture and surface blood vessel pattern to produce composite pathological-functional maps. Sufficient data for localization as well as identification of both pathological and functional architecture could be conveyed in a single image. CONCLUSIONS: Cortical maps generated with intrinsic signal imaging can combine topographic and localizing information about normal functional architecture and interictal and ictal onset zones with extremely high spatial resolution. These maps may be useful in guiding surgical resections and multiple subpial transections to minimize unnecessary damage to functional brain surrounding neocortical pathology. PMID- 15821370 TI - Mother, the divine power. PMID- 15821371 TI - Milk may protect against bowel cancer. PMID- 15821372 TI - Anaesthesia for cleft lip surgery--challenge in rural Nepal. AB - Cleft lip and palate is quite common in Nepalese population residing in hilly areas and interior of the country. Nepalese doctor with limited resources and fund and International aided team of fully equipped doctors from developed world conduct operative procedure in the centre or district hospital setup quite frequently. But very few patients will come to the hospital for treatment as it is not an emergency for them. Anaesthesia for cleft lip and palate surgery carries a high risk and difficulty in airway management. Associated congenital anomalies further complicate the management. Still a team of Nepalese physicians are providing this service in a field. How to provide a safe anaesthesia was a main concern for an anaesthesiologist with a limited supply. During the last three years we have anaesthetized safely four hundred children age between 2 months to 14 years. The team worked at an altitude ranging from 100 feet to 10,000 feet of Nepal in a temporary erected operation room of the village. Most of the operation was conducted under total Ketamine anaesthesia and general anaesthesia with halothane. In our context we found Ketamine anaesthesia was safe and effective anaesthetic technique in simple clefts in children more than 2 years. This technique reduces the cost by minimizing the supply to be carried in the field. PMID- 15821373 TI - A study of metopic sutures in human skulls. AB - The present study has been carried out in fifty-one adult human skulls for metopic sutures in the department of anatomy, Nepalgunj Medical College, Chisapani. Metopic sutures were found in six skulls (11.46%) This study showed higher incidences of incomplete metopic sutures (7.84%). One skull showed double incomplete sutures (one to right of bregma and other in the middle of the frontal bone in the mid line), which was of, a very rare variety seen in frontal bone of (1.96%), three skulls showed incomplete sutures (3.92%). Though the number of the skulls was small, the study showed a rare morphological variety, which was well correlated with earlier workers. PMID- 15821374 TI - Cholesterol-lowering effect of organosulphur compounds from garlic: a possible mechanism of action. AB - OBJECTIVES: Hyperlipidemia constitutes a major etiopathological factor for atherosclerosis. The medicinal value of garlic is best known for its lipid lowering effects and antiatherogenic effects. The mechanism by which lipid soluble organosulphur compounds from garlic reduce plasma lipids has not been fully investigated. The author had previously shown that the hepatic activity of beta-hydroxy-beta-methylglutaryl-CoA (HMG-CoA) reductase, the rate limiting enzyme in cholesterol biosynthesis and the incorporation of radiolabeled (1, 2 14C), acetate into hepatic free and esterified cholesterol was significantly decreased in rat treated with garlic derived organosulphur compounds. We hypothesised that the antiatherogenic effect of the organosulphur compounds may be attributed to the formation of protein internal disulphide and thus inactivation of thiol (-SH) group enzymes such as HMG-CoA reductase and the multienzyme complex of fatty acid synthesis. The objective of the present study is to elucidate the inhibitory mechanism by in vitro studies. METHOD: Lipid soluble organosulphur compounds from garlic were treated in vitro with Luke's cysteine reagent (representing the thiol (-SH) group of enzymes) and the interaction products were separated by paper chromatography. RESULT: The result indicated that the organosulphur compounds were capable of interacting with the thiol (-SH) group of cysteine and thus forming cysteine derivatives. CONCLUSION: The antiatherogenic effects of these organosulphur compounds can be attributed to such reactions that inhibit HMG-CoA reductase and other lipogenic enzymes. The anticarcinogenic effects of these compounds may also be due to inhibitory reactions on enzymes that activate carcinogens. PMID- 15821376 TI - Structural changes in ocular trauma and the visual outcome. AB - This is the presentation of one year prospective study on the anatomical structural lesions in ocular traumas with their visual outcome among those cases admitted and treated at Nepal Eye Hospital, Kathmandu. The changing life style of the people and environmental changes around with the changing nature of the causative objects on eye injuries have been observed for causing more serious structural lesions or complications including loss of vision. PMID- 15821375 TI - Barriers to up take cataract surgery in Gandaki Zone, Nepal. AB - OBJECTIVE: Cataract though avoidable remains as leading cause of blindness in Nepal. Though, Himalaya Eye Hospital rendering high quality surgical service through its base hospital and out reach service in Gandaki Zone having cataract surgical rate is below 1500 per million. Therefore, it is mater of interest to investigate the reason and the finding would be helpful to plan the future cataract programme to CSR to desirable level, if we know the reason(s) that forced them living as blind in community and not seeking available surgical services. METHOD: All person with cataract affecting their visual acuity to <6/60 (corrected) in either eye found in stratified cluster design sampling were enrolled in the study. They were interviewed with structured questionnaire investigating their knowledge and attitude of their visual impairment and barriers for not seeking cataract surgical service to date. RESULTS: A total of 303 eligible subjects were enrolled in the study. Out of them 57.8% of people knew about the cataract and they also think their vision impairment is due to cataract. But 56.11% of people were not aware their vision will improve. Surgical camp attendant or operated patients were found to be effective media to spread the message that the cataract is curable. Among the barriers: fear for surgery 31.1%, lack of attendant and geographical distance 32.8%, financial 24.1% and other clinical reason were found to be 12% as a reason for not seeking available surgical service. CONCLUSION: The cataract service marketing, eye health education disseminating the information on cataract and its curability is not found to be adequate. Therefore, the future cataract programme should give more concentration on creating awareness on cataract and subsidizing the available surgical service and service need to be brought to more proximity for the needy poor people. PMID- 15821377 TI - Backache prevalence among groups with long and normal working day. AB - OBJECTIVES: to describe the prevalence of backache among groups with long and normal working day. DESIGN: A cross sectional study of prevalence of backache among groups with long and normal working day. MATERIAL AND METHOD: Among the 19 wards of Kirtipur municipality, ward no. 11 was selected as a random cluster. 64 households of different socioeconomic status were selected at random. Total of 64 individuals from these households were selected in the study. CONCLUSION: The backache was found to be 62.5% prevalent. 75.8% of individuals in long working day category were found to be suffering from backache whereas 48.4% in normal working day group were suffering from it. PMID- 15821378 TI - Stripping without a stripper. AB - Varicose veins afflict some 10 to 20% of the population in the west. It is believed to be less prevalent in the developing nations. Though there are absolute indications of surgery for varicose veins, people in Nepal come for treatment of the varicose veins either to join the army or to work abroad. Not all hospitals in Nepal are equipped with a vein stripper. In KMCTH, we studied 29 patients who had their long Saphenous veins stripped either with a stripper or by tunnelling around the Saphenous vein. We found that the long Saphenous vein could be stripped via the tunnelling method with an additional incision above the knee joint which is cosmetically accepted by our patients Moreover the patients were much satisfied because of minimal or no bruises in the post operative period as compared to those who had the long Saphenous vein stripped with a stripper. PMID- 15821379 TI - A comparison of haemodynamic response with pethidine vs. butorphanol in open cholecystectomy cases. AB - OBJECTIVE: The present study was undertaken to compare the haemodynamic responses with pethidine vs. Butorphanol intraoperatively in open cholecystectomy cases in KMCTH. METHOD: In this randomized study, all together 40 patients undergoing routine cholecystectomy surgery were included. Group A received Pethidine 1 mg/kg and Group B received Butorphanol 0.04 mg/kg intraoperatively. Heart rate and blood pressure were recorded before injection of the drug, after injection, before intubation, after intubation, before skin incision, after incision, before extubation and after extubation data analysis was done using independent sample t test. RESULT: Our study showed no statistical significance in haemodynamic responses with either pethidine or Butorphanol in open cholecystectomy cases. CONCLUSION: Both drugs appear equally good analgesics in our study. PMID- 15821380 TI - Post legalisation challenge: minimizing complications of abortion. AB - Abortion has been legalized in Nepal since September 2002 by 11th amendment to the Muluki Ain. The present study was conducted in Paropakar Shree Panch Indra Rajya Laxmi Devi Maternity Hospital to assess the magnitude of induced abortion, its causes and the types of complications, in the post legalization phase. Prospective descriptive analyses of the patients who were admitted with history of induced abortion from 16th Dec 2003 to 13th March 2004 was carried out. A total of 305 cases of abortion complications were admitted during the three-month study period, which is 39.7% of the total gynaecological admissions (768). Of these 31 (10.25%) patients had history of induced abortion. Half of the induced abortion cases (52%) were of age group 21-29 yrs and 42% had three or more children. 39% of the cases had history of induced abortion at more than 12 weeks and almost half of the cases (48%) had history of family planning. The most common reason for seeking abortion was too many children (59%) followed by illegitimate pregnancy (16%). Twenty-one patients gave history of abortion being performed by doctors and the most common method used was D and C (75%). 77% of cases presented as incomplete abortion and one case presented with uterine perforation, bowel injury and peritonitis. Twenty patients had evacuation under sedation while five had manual vacuum aspiration (MVA); one patient required laparatomy. In two third of the patients intravenous fluid and antibiotics were used. Four patients required blood transfusion. Abortion complications constitute almost 40% of the total gynaecological admissions. Ten percent of the abortion cases had history of induced abortion. Medical persons, mainly doctors, performed most of the cases of induced abortion and D and C was the most commonly used method. However the patients had faced various types of complications. Untrained provider, resulting in serious life threatening injuries, performed more than a third of the cases of induced abortion at more than twelve weeks gestation. This points to the need for improved monitoring of the quality of services provided, and adherence to the criteria set by the procedural order. PMID- 15821381 TI - Abdominal tuberculosis--a study of 25 cases. AB - Abdominal tuberculosis is one of the common extra pulmonary tubercular infections. Its clinical presentation is protean and it has diagnostic dilemma, as most of the investigations are non-specific and less sensitive. Therefore this study was undertaken to define the most suggestive clinical features of abdominal tuberculosis, to find out the efficacy of most commonly available investigations and also to evaluate the response of abdominal tuberculosis to conventional antitubercular therapy. Twenty five cases of either sex were studied in department of gastroenterology in BSMMU (earlier IPGMR), Dhaka from January 2001 to June 2003. All patients were clinically evaluated and were investigated by available tests. Eighteen patients were diagnosed by investigations (One by detecting AFB, 9 with caseating granuloma in biopsy specimen and 8 with suggestive radiological findings). Diagnosis of 7 patients were made from the common clinical features (fever, weight loss, altered bowel habit, abdominal pain and distension, positive non-specific findings and from response by antitubercular therapy. All patients received conventional 9 month anti tubercular treatment with Rifampicin, Isoniazide and Pyrazinamide and were followed up clinically during and one year after completion of treatment. All patients were improved with minimum side effects of drug. Therefore, it is observed that a representative tissue biopsy (when approachable) and radiological findings are good method of diagnosis of abdominal tuberculosis. Strongly suggestive clinical features with positive non specific investigation findings are also an indication for anti tubercular treatment in all endemic countries like Nepal, Bangladesh and India. Treatment of abdominal tuberculosis is in no way different from that of other conventional anti-TB therapy. PMID- 15821382 TI - Smoking women and their lung function tests. AB - Smoking amongst women has become significant as the number of smoking women is increasing gradually in this developing society. It affects the lungs to produce Chronic Obstructive Pulmonary Disease (COPD). Present study was conducted over 100 smoker's women and 100 non-smoker women in the age group of 30-40 Years. Three Lung Function Tests--FEVI, FVC and PEFR were employed to all smoking and non-smoking women. It was observed that all the above mentioned three parameters of lung function tests were reduced significantly among smoker women as compared to non-smoker women. These reduced parameters of lung function test among heavy smokers are suggestive of chronic obstructive lung diseases. PMID- 15821383 TI - Hereditary spherocytosis. AB - Hereditary spherocytosis is a congenital haemolytic anaemia due to defect in spectrin-a RBC membrane protein and is transmitted as autosomal dominant. Due to this defect there is presence of characteristic spherical cell in peripheral blood smear and osmotic fragility is increased. Haemolytic anaemia, reticulocytosis, jaundice and splenomegaly are present. This article reports a case of a 9 year old boy who presented with a history of prolonged jaundice since the age of 4 years and recurrent pain in the right upper quadrant of abdomen. Clinical examination revealed jaundice, enlarged liver and marked splenomegaly. Investigations confirmed the diagnosis of hereditary spherocytosis by the presence of spherocytes in blood smear, raised reticulocytes and increased osmotic fragility. The patient was subjected to splenectomy after vaccination against coccobacillus and was discharged after proper advice and on post splenectomy antibiotic prophylaxis. PMID- 15821384 TI - B-Lynch brace suture: an effective method of conservative surgical management for placenta increta. AB - Placenta accreta is defined as an abnormal adherence, either in whole or in part, of the afterbirth to the underlying uterine wall. Placenta increta occurs when the placenta invades deeply into the myometrium. Placenta increta is a life threatening condition. We report a case of placenta increta managed by unilateral uterine artery and ovarian artery ligation followed by B-Lynch Brace suturing of the uterus to control bleeding from the placental bed. PMID- 15821385 TI - Geographic origin of Nepali doctors. AB - Though the history of in-country training of doctors in Nepal is not long, Nepal had started training doctors in abroad long ago. This is probably the first paper of its kind to correlate the developmental and ecological region to the country of training of Nepali doctors. This retrospective analysis reveals that nearly 38% doctors are trained in India, 22% each from former USSR and Nepal, 10% from Bangladesh, and 2.5% from Pakistan. Other countries contribute very few in the list. Nearly 2/3rd of the doctors represent the central developmental region and most of them are from Kathmandu valley. Ecologically mountain and hills are in great minority compared to Kathmandu valley and Terai. Interestingly training in former USSR shows a bit wider base regarding the origin in terms of developmental region. And Nepal has a clear broad base both in terms of developmental and ecological regions. As most of the doctors among Nepal trained ones are from IOM, the role of IOM way of selecting medical students need a deeper look into it. PMID- 15821386 TI - ICTs application for better health in Nepal. AB - Technologies with the ability to send information in a fast, efficient and cheap fashion--such as the Internet-can provide dramatic improvements in access to information, advice and care. This article explores the strengths and weaknesses of Internet to augment traditional health services and supply new ones. In doing so, it presents concrete cases in the developing world, with reference to Nepal, where Internet is being used for health-related activities--ranging from patient/doctor consultation through database services, to the management of epidemics. PMID- 15821387 TI - Evidence Based Medicine (EBM). PMID- 15821388 TI - Study of elective students. PMID- 15821389 TI - Internship: in retrospect. PMID- 15821390 TI - Cardiovascular disease in HIV-positive patients. PMID- 15821391 TI - Distinct patterns of HIV-specific memory T lymphocytes in HIV-exposed uninfected individuals and in HIV-infected patients. AB - BACKGROUND: Repeated exposure to HIV is not always associated with infection and multiple cohorts of HIV-exposed but seronegative individuals (ESN) have been described. HIV-specific CD4 and CD8 T lymphocytes are detected both in HIV patients and in ESN; we verified whether different patterns of HIV-specific memory T lymphocytes would be detected in individuals in whom exposure to HIV results or does not result in infection. METHODS: Gag-specific T cells were analysed in 15 ESN, 14 HIV patients, and 15 healthy controls using extensive flow cytometry analysis. RESULTS: Data confirmed that gag-specific T lymphocytes are present in ESN. Gag-specific T cells mainly secrete interleukin-2 in ESN and interferon-gamma in HIV patients. In addition the CD4/CD8 and the memory/naive ratios are altered, central memory (45RA-/CCR7+) CD4 and CD8 T lymphocytes are more abundant, and terminally differentiated (45RA+/CCR7- and 27-/28-) CD8 T lymphocytes are augmented in ESN individuals. CONCLUSIONS: Exposure to HIV occurs in high risk seronegative individuals; the observation that naive cells and CM are skewed in ESN indicate that this exposure is robust enough to modulate the CM/EM ratio. The increase in late effectors and in natural killer cells seen in ESN suggests a role for these cells in preventing actual infection. PMID- 15821392 TI - Disease progression in macaques with low SIV replication levels: on the relevance of TREC counts. AB - BACKGROUND: An attenuated immunodeficiency virus has been long considered innocuous. Nevertheless, converging data suggest that low levels of viral replication can still provoke AIDS. Pathogenesis of these attenuated infections is not understood. OBJECTIVES: To determine the pathogenicity of a long-term attenuated infection and to delineate T-cell dynamics during such an infection. METHODS: This is a cross-sectional study of 12 rhesus macaques infected with SIV Delta nef for 8 years. We evaluated apoptosis (annexin V), activation (HLA-DR, Ki67), and newly generated T cells (TCR excision circle: TREC). RESULTS: Infection with SIV Delta nef induced pathological CD4 T-cell depletion after 8 years of infection. Virus replication and CD8 T-cell activation positively correlated with the rate of disease progression. The frequency of TREC within CD8+CD45RA+ cells increased in SIV Delta nef-infected animals compared to age matched non-infected controls. Moreover, in the cohort of infected animals, TREC+CD45RA+CD4+ T-cell counts correlated strongly with non-progression to AIDS. The animal with the lowest rate of disease progression exhibited a 115-fold increase in TREC+CD45RA+CD4+ T-cell counts compared to age-matched non-infected controls. In contrast, the animal showing the fastest rate of progression to AIDS displayed 600-fold lower TREC+CD45RA+CD4+ T-cell counts compared to age-matched non-infected controls. CONCLUSIONS: Our results suggest that the thymus plays a major role in the pathogenesis of an attenuated SIV infection and that a sustained thymic output could maintain CD4 T-cell homeostasis in the context of low viral loads. PMID- 15821393 TI - Compartmentalization of HIV-1 in the central nervous system: role of the choroid plexus. AB - OBJECTIVES: To determine the genetic and phenotypic composition of HIV-1 found in the choroid plexus (CPx) and its relationship to virus in the brain and peripheral lymphoid tissue. DESIGN: Phenotypic and molecular comparisons of HIV-1 found in autopsy brain, CPx, and spleen tissues. METHODS: HIV-1 was co-cultured from matched postmortem brain (basal ganglia), CPx, and spleen tissues of AIDS patients with and without HIV-associated encephalitis and dementia. Viral phenotypes were determined by infection of monocyte-derived macrophages, MT-2 or co-receptor-specific cell lines. Viral env and pol sequences were determined from genomic DNA isolated directly from tissues or co-cultures, and phylogenetic comparisons were performed. RESULTS: CCR5-utilization was the most prevalent viral tropism found in all tissues, although spleen isolates also displayed CXCR4 usage. Viruses isolated from CPx consisted of both peripheral and brain-like virus, but were more related phenotypically and genetically to those found in the brain. Mutations found in the pol gene that could confer drug resistance to brain and CPx isolates were similar to those found in the periphery. CONCLUSION: The CPx contained replication-competent virus that was most similar, although distinct, from that found in the brain. It also contained some viruses with high similarity to those of peripheral origin. Compartmentalization of viral env and pol sequences indicated that differential selective pressures exist in each tissue examined. These studies suggest that the CPx may provide an environment that promotes the evolution of drug-resistant strains with central nervous system tropism, although it is unlikely to be a reservoir for archival HIV-1 variants. PMID- 15821394 TI - Atazanavir plus ritonavir or saquinavir, and lopinavir/ritonavir in patients experiencing multiple virological failures. AB - OBJECTIVE: To evaluate atazanavir/ritonavir (ATV/RTV) (300/100 mg) once daily, atazanavir/saquinavir (ATV/SQV) (400/1200 mg) once daily, and lopinavir/ritonavir (LPV/RTV) (400/100 mg) twice daily, each with tenofovir (300 mg) once daily and a nucleoside reverse transcriptase inhibitor in treatment-experienced HIV-infected patients. METHODS: Randomized, open-label, 48-week multicenter trial of 358 randomized adult patients who had failed two or more prior HAART regimens with baseline HIV RNA > or = 1000 copies/ml and CD4 cell count > or = 50 x 10(6) cells/l. RESULTS: The primary efficacy endpoint [plasma HIV RNA reduction assessed by time-averaged difference (TAD)] was similar for ATV/RTV and LPV/RTV [TAD 0.13; 97.5% confidence interval, -0.12 to 0.39] at 48 weeks. Mean reductions from baseline for ATV/RTV and LPV/RTV were comparable at 1.93 and 1.87 log10 copies/ml, respectively. Mean CD4 cell count increases were 110 and 121 x 10(6) cells/l for ATV/RTV, and LPV/RTV, respectively. The efficacy of ATV/SQV was lower than LPV/RTV by both these parameters. Declines in total cholesterol and fasting triglycerides were greater with ATV/RTV and ATV/SQV than with LPV/RTV (P < or = 0.005). Lipids in the LPV/RTV arm at week 48 generally increased from baseline. Lipid-lowering agents were used more frequently in the LPV/RTV arm than in the ATV arms (P < 0.05 versus ATV/RTV), as were antidiarrheal agents (P < or = 0.04 versus both ATV treatments). No new or unique safety findings emerged. CONCLUSIONS: ATV boosted with RTV is as effective and well tolerated as LPV/RTV in treatment-experienced patients, with a more favorable impact on serum lipids. Pharmacokinetically enhanced ATV provides a suitable choice for therapy of treatment-experienced HIV-infected patients. PMID- 15821395 TI - High virological failure rate in HIV patients after switching to a regimen with two nucleoside reverse transcriptase inhibitors plus tenofovir. AB - BACKGROUND: Regimens with two nucleoside analogue reverse transcriptase inhibitors (NRTI) plus tenofovir DF have been associated with a high failure rate when administered as first line therapy. Little is known about patients with undetectable viral loads who are switched to these regimens. METHODS: A post-hoc review of the virological outcomes at 24 weeks of patients who switched from a successful (< 50 copies/ml) highly active antiretroviral therapy regimen to a tenofovir plus two NRTI combination. RESULTS: Fifty-five patients started a two NRTI plus tenofovir regimen mostly because of previous toxicity/intolerance of the original drugs (74%). After 24 weeks, only 17 patients (31%) remained virologically suppressed. Patients with a regimen including a didanosine plus tenofovir-based regimen had significantly poorer outcomes than those on other combinations (success rate 5 versus 47.1%, P = 0.001). In contrast, patients on a regimen including zidovudine plus tenofovir showed a trend towards a better outcome (75 versus 27%, P = 0.083). Multivariate analysis confirmed the combination of didanosine plus tenofovir as the only variable associated with a higher rate of failure (odds ratio 17.7; 95% confidence interval 2.1-147; P = 0.007). Patients with previous reverse transcriptase mutations presented virological failure in all cases. At failure a new pattern, including the K65R mutation with M184V or thymidine analogue mutations, was observed. CONCLUSIONS: Even in patients with suppressed viraemia, a two NRTI plus tenofovir regimen is associated with a high virological failure rate, but significant variations are found depending on the nucleosides included. PMID- 15821396 TI - Early exclusive breastfeeding reduces the risk of postnatal HIV-1 transmission and increases HIV-free survival. AB - OBJECTIVES: The promotion of exclusive breastfeeding (EBF) to reduce the postnatal transmission (PNT) of HIV is based on limited data. In the context of a trial of postpartum vitamin A supplementation, we provided education and counseling about infant feeding and HIV, prospectively collected information on infant feeding practices, and measured associated infant infections and deaths. DESIGN AND METHODS: A total of 14 110 mother-newborn pairs were enrolled, randomly assigned to vitamin A treatment group after delivery, and followed for 2 years. At baseline, 6 weeks and 3 months, mothers were asked whether they were still breastfeeding, and whether any of 22 liquids or foods had been given to the infant. Breastfed infants were classified as exclusive, predominant, or mixed breastfed. RESULTS: A total of 4495 mothers tested HIV positive at baseline; 2060 of their babies were alive, polymerase chain reaction negative at 6 weeks, and provided complete feeding information. All infants initiated breastfeeding. Overall PNT (defined by a positive HIV test after the 6-week negative test) was 12.1%, 68.2% of which occurred after 6 months. Compared with EBF, early mixed breastfeeding was associated with a 4.03 (95% CI 0.98, 16.61), 3.79 (95% CI 1.40 10.29), and 2.60 (95% CI 1.21-5.55) greater risk of PNT at 6, 12, and 18 months, respectively. Predominant breastfeeding was associated with a 2.63 (95% CI 0.59 11.67), 2.69 (95% CI 0.95-7.63) and 1.61 (95% CI 0.72-3.64) trend towards greater PNT risk at 6, 12, and 18 months, compared with EBF. CONCLUSION: EBF may substantially reduce breastfeeding-associated HIV transmission. PMID- 15821397 TI - Prevalence of and factors associated with HIV-1 infection among drug users visiting treatment centers in Tehran, Iran. AB - OBJECTIVES: This study was conducted to identify the prevalence of and potential risk factors for HIV-1 infection among drug users visiting drug treatment centers in Tehran, Iran. DESIGN: Cross-sectional quantitative study. METHODS: Between October 2003 and May 2004, drug users who visited three public drug treatment centers in Tehran were interviewed and information about their sociodemographics, drug use characteristics, incarceration history, sexual history, and HIV/AIDS knowledge and risk perception were collected. Specimens of oral mucosal transudate were then collected from participants to be tested for HIV-1 antibodies. Logistic regression analysis was conducted on the association between the demographic and behavioral factors with HIV-1 infection. RESULTS: Overall, 611 (588 male and 23 female) drug users participated in the study. Among male injectors with HIV-1 prevalence of 15.2%, a history of shared injection inside prison [adjusted odds ratio (OR), 12.37; 95% confidence interval (CI), 2.94 51.97] was the main factor associated with HIV-1 infection. Among those who reported no history of injecting drug use, HIV-1 prevalence was 5.4%, and lack of condom use during sex was significantly associated with the infection (adjusted OR, 3.42; 95% CI, 1.25-9.36). CONCLUSIONS: HIV-1 infection is already prevalent among drug users in Tehran, Iran and shared injection inside prison has been revealed to be a particular risk factor for HIV-1 infection among injecting drug users. Harm reduction programs which have been started in Iran should be urgently expanded particularly in correctional settings and strengthened by condom use promotion to prevent sexual acquisition or transmission of HIV-1 among drug users. PMID- 15821398 TI - Rising incidence and prevalence of orphanhood in Manicaland, Zimbabwe, 1998 to 2003. AB - OBJECTIVE: To quantify and describe orphan incidence in Manicaland, eastern Zimbabwe. DESIGN: Open cohort study. METHODS: Statistical analysis of data on 13,740 and 10,308 children, aged 0-14 years, enumerated in household censuses in four socio-economic strata, 1998-2000 and 2001-2003, and 10,184 children seen in both censuses (74% follow-up). RESULTS: Prevalence of all forms of orphanhood increased. The overall rate of losing a parent amongst non-orphans was 27.5 per 1000 person-years (py). Paternal orphan incidence (20.2 per 1000 py) was higher than maternal orphan incidence (9.1 per 1000 py) and maternal orphans lost their fathers at a faster rate than paternal orphans lost their mothers. Paternal and maternal orphan incidence increased with age. Incidence of maternal orphanhood and double orphanhood amongst paternal orphans rose at 20% per annum [incidence rate ratio (IRR) = 1.20; 95% CI, 1.06-1.35] and 71% per annum (IRR = 1.71; 95% CI, 1.25-2.33), respectively, 1998-2003, but incidence of paternal orphanhood and double orphanhood amongst maternal orphans were unchanged. For 82% of children with a parent who died, the parent was HIV-positive at baseline. More new paternal and double orphans--but not new maternal orphans--than non-orphans had left their baseline household. Mortality was higher in orphans than non-orphans with the highest death rates observed amongst maternal orphans. CONCLUSIONS: Orphan incidence and prevalence are high and increasing due to HIV in eastern Zimbabwe. Orphan incidence patterns differ from orphan prevalence patterns and need to be understood if support programmes are to assist children during periods of high vulnerability. PMID- 15821399 TI - Risk factors for late HIV diagnosis in French Guiana. AB - Risk factors for delayed HIV diagnosis in French Guiana were studied in 1952 patients between 1992 and 2003. At the time of diagnosis, 30% of patients had less than 200 CD4 lymphocytes/mm3; age, male sex, and foreign nationality were independently associated with a low CD4 cell count. The availability of highly active antiretroviral therapy was not associated with an earlier HIV diagnosis. Promoting HIV information and testing should be done in several languages to reach minorities. PMID- 15821400 TI - Change in atherosclerosis progression in HIV-infected patients: ANRS Aquitaine Cohort, 1999-2004. AB - This study reported the changes in carotid intima-media thickness (IMT) during a 36-month period in 233 HIV-infected patients. Median IMT increased in the first 12 months and then decreased by month 36. The prevalence of treatment with lipid lowering agents and protease inhibitor-free highly active antiretroviral therapy regimens increased, whereas smoking prevalence decreased. The progression of atherosclerosis in HIV-infected patients can be controlled. The impact of individual measures to reduce the cardiovascular risk should be evaluated further. PMID- 15821402 TI - A conserved HLA B13-restricted cytotoxic T lymphocyte epitope in Nef is a dominant epitope in HLA B13-positive HIV-1-infected patients. AB - We report on the first HLA B13-restricted minimal cytotoxic T lymphocyte (CTL) epitope RQDILDLWI (RI9, amino acids 106-114 in HIV-1 Nef). In most patients the frequency of RI9-specific CTL exceeded the number of CTL against other epitopes, indicating that RI9 is a dominant epitope in HLA B13-positive patients. Targeting this conserved Nef epitope may be an important factor for the published association of HLA B13 with a favourable course of HIV-1 infection. PMID- 15821401 TI - Panel of prototypical infectious molecular HIV-1 clones containing multiple nucleoside reverse transcriptase inhibitor resistance mutations. AB - We have created a panel of recombinant HIV-1 infectious clones containing common patterns of reverse transcriptase (RT) mutations responsible for resistance to each of the currently available nucleoside reverse transcriptase inhibitors (NRTI), and we have submitted the panel to the National Institutes of Health AIDS Research and Reference Reagent Programme. Testing the activity of new antiretroviral compounds against this panel of drug-resistant clones will determine their relative activity against many clinically relevant NRTI-resistant viruses. PMID- 15821403 TI - Lower incidence and severity of cytomegalovirus-associated immune recovery uveitis in HIV-infected patients with delayed highly active antiretroviral therapy. AB - An analysis of clinical records after the introduction of highly active antiretroviral therapy (HAART) in Mexico showed a lower prevalence and severity of immune recovery uveitis when HIV-infected patients started HAART after completing cytomegalovirus retinitis treatment than when treatments were concomitant. The findings suggest a protective role of delaying HAART until cytomegalovirus retinitis is controlled. PMID- 15821404 TI - Uridine pharmacokinetics of mitocnol, a sugar cane extract. PMID- 15821405 TI - Iatrogenic Cushing's syndrome in an HIV-infected patient treated with ritonavir and inhaled fluticasone. PMID- 15821406 TI - Response to 'Single phylogenetic reconstruction method is insufficient to clarify relationships between patient isolates in HIV-1 transmission case' by Jenwitheesuk and Liu. PMID- 15821408 TI - Identifying deaths from AIDS in South Africa: an update. PMID- 15821410 TI - New capillary growth: a contributor to regression of sclerosis? PMID- 15821411 TI - Podocyte damage damages podocytes: autonomous vicious cycle that drives local spread of glomerular sclerosis. AB - PURPOSE OF REVIEW: For some time, the so-called vicious cycle has been believed to underlie progression of glomerular sclerosis. This mechanism describes a circumstance when loss of some glomeruli imposes injurious stress on the remnant glomeruli. Evidence from recent genetic approaches, however, has prompted revision of this classical view and now points toward a new direction of investigations. RECENT FINDINGS: Whereas experimental maneuvers that selectively injure mesangial cells have failed to induce glomerular sclerosis, genetic approaches that target visceral epithelial cells, or podocytes, in embryos and adult animals regularly produce glomerular sclerosis. Association between podocyte damage and glomerular sclerosis observed in many human diseases and animal models have identified podocyte injury as a common, if not universal, trigger leading to glomerular sclerosis. The process from podocyte injury to sclerosis is remarkably rapid, and the rate of progression depends upon the degree of initial podocyte injury. A single brief injurious stimulus on a podocyte activates a 'domino effect', whereby progressive damage of the initially hit podocyte spreads to involve cells that escaped the initial insult. SUMMARY: The mouse, a species highly useful for studying the function of specific gene products, is notoriously resistant to development of glomerular sclerosis in adulthood. However, recent genetic engineering in this species has overcome this disadvantage and brought about a new dimension to our understanding of the mechanisms involved in progressive glomerular sclerosis. PMID- 15821412 TI - The slit diaphragm: a signaling platform to regulate podocyte function. AB - PURPOSE OF REVIEW: The discovery of nephrin and other slit diaphragm proteins dramatically expanded our knowledge of the molecular make-up of the glomerular filter of the kidney. This review focuses on the most recent evidence elucidating the dynamic functions of the slit diaphragm and stresses the importance of slit diaphragm proteins in mediating signal transduction in the podocyte. RECENT FINDINGS: Nephrin and neph molecules form specialized cell junctions in various systems in different species. The organization of these specialized cell junctions is dependent on intracellular signaling networks. There is a rapidly growing number of identified adapter and signaling molecules that are recruited to this signaling network. These proteins help to maintain programs for cell survival, cell polarity and endocytosis. The latest examples of identified signaling proteins at the slit diaphragm are fyn, p85, and calcium/calmodulin dependent serine protein kinase. At the level of the slit insertion site, podocin seems to play a pivotal role in the establishment of a lipid-enriched signaling environment. Moreover, the protocadherin FAT1 has been identified as an organizer of actin polymerization and could therefore serve as a linker of the slit diaphragm to cytoskeletal organization. SUMMARY: From recent data, a novel concept of a highly dynamic slit protein complex is emerging. Slit diaphragm proteins are contributing to the regulation of cell polarity, cell survival and cytoskeletal organization. This concept is further supported by the fact that many clinically relevant mutations of slit diaphragm proteins interfere with signaling processes at the slit diaphragm and cause proteinuria and progressive kidney disease. PMID- 15821413 TI - New insights that link microbes with the generation of antineutrophil cytoplasmic autoantibodies: the theory of autoantigen complementarity. AB - PURPOSE OF REVIEW: Reviewed are recent discoveries that provide insights into novel mechanisms involved in the aetiology and pathology of anti-neutrophil cytoplasmic autoantibodies (ANCA) disease. RECENT FINDINGS: Gene expression profiles of circulating leukocytes from anti-neutrophil cytoplasmic autoantibody immunogenesis patients revealed high levels of proteinase 3 (PR3) and myeloperoxidase (MPO) mRNA. Combined with reports of increased expression of these proteins, it appears that increased antigen availability is a pathologic component of anti-neutrophil cytoplasmic autoantibody immunogenesis disease, which might be equally as important as the presence of anti-MPO or anti-PR3 autoantibodies. Genetic predisposition to develop anti-neutrophil cytoplasmic autoantibody immunogenesis disease may include a polymorphism in the promoter region of the PR3 gene. Signalling pathways affected by anti-neutrophil cytoplasmic autoantibody immunogenesis binding to neutrophils involve the p21 pathway. Lastly, a topic discussed at length in this review is the seminal observation that PR3-ANCA patients harbour antibodies reactive with a protein produced from PR3-antisense RNA, whose amino acid sequence has homologies with proteins from many microbes and viruses. Delineated in the Theory of Autoantigen Complementarity, it is proposed that the initiator of an autoimmune response is not the autoantigen, but instead is a protein that is 'antisense' or complementary to the autoantigen (e.g. from bacteria or PR3). SUMMARY: The progress in research efforts in the past year, including the identification of complementary proteins as a potential cause of anti-neutrophil cytoplasmic autoantibody immunogenesis, should highly impact future approaches therapeutic intervention. PMID- 15821414 TI - Complement and glomerulonephritis: new insights. AB - PURPOSE OF REVIEW: The last few years have seen a huge increase in our understanding of the role of the complement system and its regulation in glomerular disease. Our aim is to summarize the most important advances in this field. RECENT FINDINGS: The role of complement in systemic lupus erythematosus continues to be elucidated. Classical pathway components protect from the development of autoimmunity, at least in part, through their role in the clearance of apoptotic cells. In contrast, the alternative pathway plays a direct role in exacerbating glomerular injury. Anti-C1q antibodies are related to activity in lupus nephritis and recent studies have shown that they are directly pathogenic in animal models. Proteinuria, whatever the cause, may lead to tubulointerstitial injury and complement activation adds to this process. In particular, deposition of terminal components of complement in the tubular lumen contributes to interstitial myofibroblast activation. There is increasing evidence for the role of complement regulatory proteins in glomerular injury. In particular, abnormalities of factor H or of CD46 may predispose to atypical haemolytic uraemic syndrome. The control proteins also protect against injury in immune complex glomerulonephritis. SUMMARY: Advances in our understanding of the role of complement in glomerular injury point to the likely therapeutic benefits of targeting the complement system. Many new drugs are becoming available. Careful dissection of the pro and antiinflammatory effects of the complement system which the experimental models allow will assist in designing directed therapy that will avoid the detrimental effects of nonspecific systemic complement inhibition. PMID- 15821415 TI - Chronic allograft nephropathy. AB - PURPOSE OF REVIEW: Chronic allograft nephropathy is the major cause of late renal allograft loss. This disease is heterogeneous and the diagnosis is nonspecific, with both immune and nonimmune causes. Increasingly, we are able to recognize specific contributors to the disease. RECENT FINDINGS: Further understanding of chronic allograft nephropathy comes from a large study detailing the natural history of the disease, from protocol biopsies revealing subclinical cellular rejection, and from studies using C4d staining to distinguish antibody-mediated chronic rejection from nonspecific causes. Also made more clear are nonimmune mechanisms of chronic allograft nephropathy, such as the effect of decreased dosing of calcineurin inhibitors, and the concept of senescence as a mechanism of the disease. SUMMARY: Chronic allograft nephropathy is a heterogeneous disease with immune and nonimmune causes. Some features recognizable by histology and detected by other laboratory tests can help to categorize specific causes of the disease in particular cases. In addition, recent studies have contributed to our knowledge of the pathogenesis of the disease. In order to advance our understanding, we must be able to distinguish the various recognizable causes of chronic allograft dysfunction. Further research is warranted on the subset of the disease with indeterminate cause. PMID- 15821416 TI - Aldosterone and end-organ damage. AB - PURPOSE OF REVIEW: This review highlights recent clinical studies demonstrating the contribution of aldosterone to cardiovascular mortality, vascular dysfunction, and renal injury in the context of advances in our understanding of the molecular biology of aldosterone. RECENT FINDINGS: Mineralocorticoid receptor antagonism reduces mortality in patients with congestive heart failure and following myocardial infarction. Studies in animal models and in patients with congestive heart failure or hypertension indicate that aldosterone induces oxidative stress and impairs endothelial nitric oxide synthase through a mineralocorticoid receptor-dependent mechanism. Furthermore, aldosterone can cause vasoconstriction and vasodilation through rapid nongenomic mechanisms. The contribution of the nongenomic effects of aldosterone to vascular tone may depend on underlying endothelial function. In the heart and kidney, aldosterone stimulates oxidative stress and increases expression of inflammatory markers leading to fibrosis. The induction of inflammation and fibrosis appears to be both sodium and mineralocorticoid receptor dependent. The mechanisms underlying the progression from inflammation to fibrosis remain to be elucidated. Studies measuring circulating markers of collagen turnover suggest that mineralocorticoid antagonism reduces extracellular matrix turnover and cardiac remodeling in humans as well. Similarly, mineralocorticoid receptor antagonism reduces urinary albumen excretion in clinical trials in humans. SUMMARY: Aldosterone induces oxidative stress, endothelial dysfunction, inflammation and fibrosis in the vasculature, heart and kidney. While most of these effects appear to be mediated via the mineralocorticoid receptor, better understanding of the mineralocorticoid receptor-independent effects of aldosterone, the role of nonaldosterone mineralocorticoid receptor agonists, and the mechanisms involved in the progression from inflammation to fibrosis and remodeling would enable the development of new strategies to slow the progression of cardiovascular and renal disease. PMID- 15821417 TI - Molecular events in kidney ageing. AB - PURPOSE OF REVIEW: Ageing of the kidney is a problem of clinical and basic interest. The problem of renal dysfunction and end-stage renal disease is a major burden on the health system, and old donor age is a major limitation on the use of donor organs and on survival of transplanted kidneys. Moreover, stresses linked to nephropathies, postoperative stress, inflammation and allograft rejection can lead to premature senescence of renal cells thus accelerating organ atrophy. Age-related and disease or stress-related nephron loss could reflect both the limited ability of epithelial cells to repair and replicate in the face of environmental stresses, and limitations on the number of cell replications caused by telomere shortening. Therefore, elucidating cellular senescence mechanisms is relevant to kidney diseases and kidney transplantation. RECENT FINDINGS: Recent findings suggest additive effects of replicative and environmental stress-induced senescence in cellular and organ ageing. In particular, ATM/p53/p21 and Ras/p38/p16 pathways have been shown to co-contribute to the overall cellular senescence, which is caused by extrinsic and intrinsic stimuli. Moreover, the role of epigenetic factors, including protein acylation/deacetylation, chromatin remodeling or caloric restriction, is the focus of recent studies on ageing and senescence. SUMMARY: Despite significant progress, cellular senescence is still better understood in vitro than in vivo. So far, p16 remains the best marker of chronological age in the kidney, and can be considered as an indicator of premature senescence caused by stresses or disease. The beneficial effects of caloric restriction on organ ageing and the role of histone acetylation in pathologic states in rodents are of considerable interest, and deserve future studies. PMID- 15821418 TI - Screening for kidney disease in adults with diabetes and prediabetes. AB - PURPOSE OF REVIEW: Worldwide, we are facing an increasing prevalence of diabetes mellitus, the number one cause of end-stage renal disease. It is imperative that chronic kidney disease among adults with diabetes mellitus be detected early when interventions are most effective in preventing end-stage renal disease. RECENT FINDINGS: Screening tools work best when health care providers have a clear understanding of the natural history of the disease process. The use of urine albumin measurement as a screening tool for diabetic nephropathy is based on the well characterized evolution of nephropathy in type 1 diabetes mellitus. However, the evolution of chronic kidney disease among adults with type 2 diabetes mellitus appears to be more heterogeneous. In contrast to type 1 diabetes mellitus, a substantial percentage of adults with type 2 diabetes mellitus have decreased glomerular filtration rate in the absence of increased urine albumin excretion. Focusing solely on urine albumin excretion to screen for chronic kidney disease may miss a substantial number of cases in adults with diabetes mellitus. In addition, up to one-third of adults with newly diagnosed diabetes mellitus already have chronic kidney disease, which may have first developed during the prediabetic state. Screening for chronic kidney disease only after an adult develops diabetes mellitus may miss that 'early' window of opportunity in many patients. SUMMARY: To maximize the effectiveness of our screening programs, we must not only use tests with adequate sensitivity but also implement screening tests early in the disease process to prevent the progression of chronic kidney disease to end-stage renal disease. PMID- 15821419 TI - Can dietary changes reduce blood pressure in the long term? AB - PURPOSE OF REVIEW: Recent dietary guidelines for the prevention and treatment of hypertension emphasized a healthy eating pattern called the Dietary Approaches to Stop Hypertension (DASH) diet, in addition to avoidance of obesity, high salt intake, and excessive alcohol intake. Our new challenge is to examine the long term efficacy and effectiveness of dietary change, including the DASH diet, on blood pressure in the general population, or in a high-risk population. We reviewed recent short-term and long-term studies, both observational and interventional, which investigated whether dietary change can reduce blood pressure. RECENT FINDINGS: The efficacy of several nutrients has been investigated in short-term trials (e.g. polyunsaturated fatty acids, including omega-3 fatty acids, and trans fatty acids), but no substantial effect of these nutrients on blood pressure has been proved, except for dietary fiber. The PREMIER trial showed the effectiveness of implementing the DASH diet for 6 months in addition to the established recommendations on lifestyle modification, but other long-term trials of dietary modification did not show sufficient blood pressure reduction. However, long-term observational studies have shown that a higher intake of fruit and vegetables and a lower intake of red meat (except fish) can prevent higher blood pressure increase with age. SUMMARY: More effective programs of dietary modification are needed to achieve greater reduction of blood pressure. Long-term studies of population-wide dietary interventions to decrease the blood pressure level of a whole population are also needed. PMID- 15821420 TI - Early life determinants of adult blood pressure. AB - PURPOSE OF REVIEW: While treating high blood pressure in middle age is beneficial in terms of reducing the occurrence of cardiovascular disease, treated and well controlled hypertensive adults still have a substantial excess mortality and reduced survival compared with normotensives. Therefore, identification of the means of preventing hypertension in earlier life is an important objective. There is increasing evidence that adult blood pressure is determined by a range of characteristics from the intrauterine period, through infancy and childhood. The purpose of this review is to provide a summary of the current evidence concerning the early life determinants of adult blood pressure. RECENT FINDINGS: Children from poorer socioeconomic positions, those whose mothers experience pregnancy induced hypertension, those whose mothers smoke throughout pregnancy, those with low birthweight, who are not breast-fed, who have high sodium diets in infancy and who are obese in childhood or adolescence tend to have higher blood pressure in adulthood. However, the mechanisms linking these early life factors to later blood pressure and the most appropriate means of preventing adult hypertension by intervening in early life are unclear. SUMMARY: There is clear evidence that early life factors are important determinants of adult blood pressure. However, there is a need for randomized trials with sufficient resources for long-term follow-up to assess the effects that interventions such as preventing pregnancy induced hypertension, reducing maternal smoking, increasing breast-feeding, reducing salt consumption in infancy and preventing childhood obesity have on adult blood pressure and cardiovascular disease. PMID- 15821421 TI - Are small changes in serum creatinine an important risk factor? AB - PURPOSE OF REVIEW: Serum creatinine levels are strongly associated with longitudinal risk for cardiovascular disease and mortality. Recent studies addressed whether worsening renal function - defined by small increases in creatinine - is independently associated with adverse outcomes. This review evaluates the recent literature on worsened renal function as an independent risk factor. RECENT FINDINGS: Studies have evaluated worsening renal function as a predictor of cardiovascular outcomes and mortality in three settings: cardiac surgery patients, hospitalized heart failure patients, and ambulatory coronary artery disease patients. Small creatinine changes following cardiac surgery were strongly associated with mortality risk. One study found a J-shaped association between 48 h post surgery creatinine change and 30-day mortality risk. Compared with patients with creatinine decreases of 0-0.3 mg/dl, patients with creatinine increases less than 0.5 mg/dl had a twofold adjusted mortality risk and those with creatinine increases of at least 0.5 mg/dl had a nearly sixfold mortality risk; surprisingly those with decreases over 0.3 mg/dl had a twofold adjusted risk. Worsening renal function was also a strong predictor of mortality for hospitalized heart failure patients independent of baseline creatinine; the magnitude of creatinine rise appeared to be linearly associated with mortality risk. However, one study found no independent association between worsening renal function and cardiovascular or mortality risk over longer follow-up. SUMMARY: Acute elevations in serum creatinine had a linear association with increased risk for adverse outcomes among patients hospitalized for cardiac surgery or heart failure. Future studies should determine interventions to prevent and treat in hospital worsening renal function to reduce the risk for adverse outcomes. PMID- 15821422 TI - What is the evidence that microalbuminuria is a predictor of cardiovascular disease events? AB - PURPOSE OF REVIEW: This review describes recently published studies evaluating the association between microalbuminuria and the development of cardiovascular disease events either in the presence of diabetes or hypertension, or in the population as a whole. RECENT FINDINGS: Prospective studies confirm that microalbuminuria is predictive, independently of classical risk factors, of cardiovascular disease events and all-cause mortality within groups of patients with diabetes or hypertension and in the general population. However, these studies suggest that levels of albuminuria below the conventional cutoff point definition of microalbuminuria are significantly associated with cardiovascular morbidity and mortality. The pathophysiological mechanism underyling this association is still uncertain. Data from recent intervention studies suggest that treatment with angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers, as well as intensive multi-factorial intervention including behaviour modification and targeted pharmacotherapy in patients with microalbuminuria, offers significant reduction in cardiovascular and renal morbidity in people with albuminuria. SUMMARY: Future absolute risk prediction scores for primary cardiovascular events could include microalbuminuria as a modifiable risk factor. The association between levels of albuminuria and cardiovascular outcomes in individuals within the normoalbuminuric range questions the current categorical definition of microalbuminuria. Intensive multifactorial interventions, including the use of agents that affect the renin angiotensin pathway, are effective in reducing cardiovascular risk in patients with microalbuminuria and diabetes or hypertension. PMID- 15821424 TI - Bibliography. Current world literature. Renal immunology and pathology. PMID- 15821425 TI - Muscle strength, body composition, hormones, and aging. PMID- 15821423 TI - Folic acid supplementation to prevent adverse events in individuals with chronic kidney disease and end stage renal disease. AB - PURPOSE OF REVIEW: This review summarizes our current understanding of the role of folate in the treatment of hyperhomocysteindemia and the prevention of cardiovascular disease in patients with chronic kidney disease and end stage renal disease. Relevant papers published between 2003 and 2004 are referenced. RECENT FINDINGS: With the exception of one paper, recent therapeutic studies supported previous findings that folate therapy achieves only a modest reduction in plasma homocysteine and seldom normalizes homocysteine. Large prospective studies are under way to evaluate the causal relationship between homocysteine and cardiovascular risk. Recent work supports earlier data that suggested that homocysteine inflicts its damage by oxidative stress. A newly described consequence of hyperhomocysteindemia is DNA hypomethylation and alteration of gene expression. A recent study in the general population suggested that while folate may lower homocysteine it does not improve endothelial function in individuals without cardiovascular disease. SUMMARY: The causes of hyperhomocysteindemia in renal failure remain obscure. The possibilities include impairment of both renal and extrarenal metabolic pathways by uraemia. Hyperhomocysteindemia is associated in some but not all studies with an increased risk for cardiovascular disease. A low homocysteine may reflect malnutrition and predict a poor outcome. Folate achieves modest reductions of homocysteine in some but not all studies. There are no data to support therapy with very high-dose folic acid. Hyperhomocysteindemia impairs endothelial function which is not adequately reversed by folate. PMID- 15821426 TI - Muscle stem cells and exercise training. AB - Adult skeletal muscle fibers are terminally differentiated such that hypertrophy and regeneration require resident stem cell populations. This review examines the current understanding of the factors regulating muscle stem cells in response to exercise and identify the role of these cells in the adaptive response of skeletal muscle to endurance and resistance exercise training. PMID- 15821427 TI - Can the physical environment determine physical activity levels? AB - Does your place of residence affect your level of physical activity and ultimately your weight and health? There is relatively strong evidence of association between compact development patterns and use of active travel modes such as walking and transit. There is weaker evidence of linkage between compact development, overall physical activity, and downstream weight and health effects. PMID- 15821428 TI - Summation of motor unit force in passive and active muscle. AB - Nonlinear summation of force has been observed between motor units. The complex structure of muscle suggests many reasons why this could happen. When large portions of the muscle are active, however, the nonlinearities are small, and generally explained by stretch of the common elasticity. This suggests that the type of nonlinearity observed between motor units may not be physiologically significant. PMID- 15821429 TI - The subacute effects of exercise: concept, characteristics, and clinical implications. AB - Physical exercise represents a physiological stress to the organism producing acute integrative physiological adjustments, whereas repetitive and regular sessions of physical effort provoke chronic physiological adaptations that develop throughout the weeks of training and increase exercise capacity. The present work presents and discusses the concept of subacute effects of exercise, those physiological responses occurring between single bouts of exercise. PMID- 15821430 TI - Energetic consequences of walking like an inverted pendulum: step-to-step transitions. AB - Walking like an inverted pendulum reduces muscle-force and work demands during single support, but it also unavoidably requires mechanical work to redirect the body's center of mass in the transition between steps, when one pendular motion is substituted by the next. Production of this work exacts a proportional metabolic cost that is a major determinant of the overall cost of walking. PMID- 15821431 TI - Damage to skeletal muscle from eccentric exercise. AB - Evidence is provided for a mechanical event as the first step in the process leading to muscle damage after a series of eccentric contractions. Aspects discussed include the decline in active tension, increase in passive tension, shift in length-tension relation, soreness, swelling, and disturbed proprioception. PMID- 15821432 TI - Differential effects of MK801 and lorazepam on heart rate variability in adolescent rhesus monkeys (macaca mulatta). AB - Previous research shows that ketamine significantly alters cardiac signal regulation in rhesus monkeys, however relatively little is known about the mechanism for this effect. In the study reported here the relative contributions of NMDA receptor activation on cardiac signal dynamics were determined by administering a specific NMDA antagonist, MK801, to rhesus monkeys. The general effects of sedation were assessed by measuring cardiac response to lorazepam, a sedative drug without direct NMDA receptor activity. Electrocardiographic signal dynamics were examined before and after I.V. administration of either MK801 (0.16 mg/kg) or lorazepam (0.48 mg/kg). Inter-beat interval time series data were analyzed in the frequency domain after Fourier transform, and a nonlinear measure of autocorrelation, the Hurst exponent (H), was derived. After MK801 administration, log [HF /Total power] increased post-infusion (M = 1.11, SD = 0.45) compared with pre-infusion values [M = -0.19, SD = 0.32, F(1,4) = 19.49, P = 0.01] while H decreased, mean pre versus post 0.52+/-S.D. 0.10 versus 0.01+/- 0.05, P = 0.0002. Lorazepam administration did not significantly alter heart rate variability measures obtained in the frequency or nonlinear domains. To our knowledge, this is the first study that has defined the effects of peripherally administered MK801 on cardiovascular dynamics in primates and establishes that peripheral administration of NMDA antagonists result in large increases the high frequency components of cardiac rhythm and increased heart rate variability compatible with MK801-associated increases in parasympathetic outflow. PMID- 15821433 TI - Antithrombotic properties of the thromboxane A2/prostaglandin H2 receptor antagonist S18886 on prevention of platelet-dependent cyclic flow reductions in dogs. AB - A potent thromboxane A2/PGH2 (TP)-receptor antagonist, S18886, was evaluated for its antithrombotic property in a dog model of acute periodic platelet-mediated thrombosis in stenosed coronary arteries with endothelial damage. After thrombosis had been obtained in 11 dogs, S18886 (300 microg/kg bolus) was administered IV. Heart rate, systemic blood pressure, and coronary blood flow were continuously recorded. Ex vivo whole blood platelet aggregation (PA), blood pH, hematocrit, platelet count, PO2, PCO2, and bleeding times were measured before and 30 minutes after administration of S18886. S18886 completely inhibited thrombosis in all dogs in approximately 5-10 minutes. No change in heart rate, blood pressure, pH, PO2, PCO2, platelet count, or bleeding time and a slight but significant elevation in hematocrit occurred. Infusion of epinephrine IV after complete inhibition of thrombosis by S18886 partially restored thrombosis in 3 of the 11 dogs. PA induced by collagen (4 microg/mL), collagen (0.25 microg/mL) plus epinephrine (1 microg/mL), collagen (1 microg/mL) plus epinephrine (1 microg/mL), ADP (40 microM) plus epinephrine (1 microg/mL), and phorbol 12-myristate 13 acetate (0.5 nM) were attenuated by 90 +/- 8% (P < 0.005), 98 +/- 2% (P < 0.05), 78 +/- 6% (P < 0.005), 70 +/- 10% (P < 0.005), and 28 +/- 8% (P < 0.05), respectively. In conclusion, S18886 is a potent platelet inhibitor that attenuates in vivo platelet-dependent thrombosis in the experimental dog model and reduces ex vivo platelet aggregation. PMID- 15821434 TI - Inducible nitric oxide synthase inhibition potentiates multiple organ dysfunction induced by endotoxin in conscious rats. AB - This study was designed to investigate the effects of inducible nitric oxide synthase (iNOS) inhibition with S-methylisothiourea (SMT) and L-N-(1-iminoethyl) lysine (L-Nil) on the endotoxemia induced by intravenous lipopolysaccharide (LPS, 10 mg/kg) in conscious rats. Arterial pressure (AP), heart rate (HR), WBC, platelets, plasma nitrite/nitrate, tumor necrosis factor alpha (TNF alpha), and biochemical factors were measured for 24 hours after LPS with or without iNOS inhibitors. RT-PCR was employed to determine the iNOS and endothelial NOS (eNOS) mRNA. Pathologic examinations of the liver and heart were performed. SMT and L Nil improved the systemic hypotension and increased the HR after LPS. These agents attenuated the LPS-induced leukocytopenia and thrombocytopenia and the increase in nitrite/nitrate. However, iNOS inhibition aggravated the LPS-induced changes in TNF alpha, all biochemical factors, and the hepatic and cardiac tissue damage. The iNOS mRNA, but not the eNOS, was reduced. Our results in conscious rats indicate that iNOS inhibition enhances the organ dysfunction and tissue damage in sepsis. The discrepancy may be attributed to the method for evaluating the sepsis and the effects of anesthesia. Further investigation is required to ensure the effects of iNOS inhibition on sepsis before iNOS inhibitors can be applied in clinical cases with sepsis. PMID- 15821435 TI - Inhibition of cGMP-dependent protein kinase reverses phenotypic modulation of vascular smooth muscle cells. AB - We have previously shown that type I cGMP-dependent protein kinase (PKG) can alter the phenotype of cultured vascular smooth muscle cells (VSMCs). Although the expression of contractile proteins in VSMCs has been shown to be modulated with the induction of PKG, experiments in which PKG inhibition brings about reduced expression of contractile markers have not been performed. To more thoroughly examine the role of PKG in the expression of contractile proteins, recombinant adenovirus containing the PKG coding sequence (AD-PKG) was used to induce gene expression and morphologic changes in adult rat aortic VSMCs. Cells expressing PKG, but not control adenovirus-infected cells, began to express a specific marker protein for the contractile phenotype, smooth muscle myosin heavy chain (SMMHC), within 48 hours of PKG induction. The morphology of the AD-PKG infected cells began to change from a fibroblastic phenotype to a spindle-shaped phenotype within 72 hours after PKG induction. The specific cell-permeable PKG inhibitory peptide DT-2, but not control peptides, reversed the biochemical and morphologic changes associated with PKG expression. These results suggest that PKG expression and activity in cultured VSMCs is capable of altering the VSMC phenotype. These data also verify the intracellular action of DT-2 and reveal uptake and dynamic properties of this PKG-inhibiting peptide. PMID- 15821436 TI - Endogenous estrogens increase postischemic hyperemia in the skin microcirculation. AB - Estrogens have been recognized as a major regulator of vascular tone and structure, particularly in the skin. The objective of this study was to investigate the effects of endogenous estrogens on the skin microcirculation. Skin blood flow was measured at the forearm at rest and during postischemic hyperemia using laser Doppler flowmetry in 32 healthy women (mean age 34.5 +/- 3.9 years) involved in an in-vitro fertilization program. Women were treated for 10 to 12 days with gonadotropin-releasing hormone agonist (total dose 40.3 +/- 3.3 mg) and human menopausal gonadotropin (1942 +/- 801 IE) or follicle stimulating hormone (2544 +/- 1071 IE) according to individual estrogen levels. Plasma estrogen levels increased from 132 +/- 90 pmol/L (36 +/- 25 pg/mL) to 8471 +/- 4386 pmol/L (2308 +/- 1195 pg/mL) during treatment (P < 0.0001). Maximal hyperemic blood flow increased from 353 +/- 81% before treatment to 516 +/- 144% after hormonal stimulation (P < 0.0001), whereas basal skin flow was not altered. This study shows that endogenous estrogens enhance the postischemic hyperemic response of the skin microcirculation. PMID- 15821437 TI - T-cadherin mediates low-density lipoprotein-initiated cell proliferation via the Ca(2+)-tyrosine kinase-Erk1/2 pathway. AB - The GPI-anchored protein T-cadherin was found to be an atypical LDL binding site that is expressed in various types of cells, including endothelial cells, smooth muscle cells, and neurons. Notably, the expression of T-cadherin was reduced in numerous types of cancers, although it was up-regulated in tumor-penetrating blood vessels, atherosclerotic lesions, and during neointima formation. Despite these intriguing findings, our knowledge of the physiological role and the signal transduction pathways associated with this protein is limited. Therefore, T cadherin was overexpressed in the human umbilical vein-derived endothelial cell line EA.hy926, the human embryonic kidney cell line HEK293, and LDL-initiated signal transduction, and its consequences were elucidated. Our data revealed that T-cadherin serves as a receptor specifically for LDL. Following LDL binding to T cadherin, mitogenic signal transduction was initiated that involved activation of PLC and IP3 formation, which subsequently yielded intracellular Ca2+ mobilization. Downstream to these early phenomena, activation of tyrosine kinase(s) Erk 1/2 kinase, and the translocation of NF kappa B toward the nucleus were found. Finally, overexpression of T-cadherin in HEK293 cells resulted in accelerated cell proliferation in an LDL-dependent manner, although cell viability was not influenced. Because LDL uptake was not facilitated by T cadherin, our data suggest that T-cadherin serves as a signaling receptor for LDL that facilitates an LDL-dependent mitogenic signal in the vasculature. PMID- 15821438 TI - Analysis of pulmonary vascular response to acute alveolar hypoxic challenge in young rabbits subjected to chronic hypoxia from birth. AB - Chronic alveolar hypoxia induces vascular changes leading to pulmonary hypertension. We investigated the role of nitric oxide synthase (NOS) on basal pulmonary artery pressure (PAP) and on changes in PAP arising from an acute alveolar hypoxic challenge (AAHC) in normoxic and chronically hypoxic young rabbits. The chronically hypoxic rabbits were raised from birth in a chamber containing a (10% O2 + 90% N2) gas mixture, whereas the normoxic rabbits were kept in room air. The age of the animals at the time of study (approximately 38 days) was not significantly different between the 2 groups of rabbits. The in vivo PAP was measured using a right heart catheterization technique while the rabbits were spontaneously breathing either a hyperoxic or a hypoxic gas. In the chronically hypoxic group, the AAHC (hypoxic gas) produced a modest increase in PAP. However, after intravenous administration of (100 mg/kg) of the NOS inhibitor, L-NAME (N-nitro-L-arginine methyl ester), a marked increase in PAP was observed when these rabbits were rechallenged with the AAHC. In contrast, in the normoxic rabbits, the AAHC produced only a small increase in PAP, even after pretreatment with L-NAME. In both groups of rabbits, L-NAME led to a significant rise in basal PAP. Using Western blot analysis, we found endothelial NOS (eNOS) protein expression to be significantly increased in pulmonary artery and right ventricular myocardium of the chronically hypoxic rabbits. These results suggest that release of nitric oxide is involved in regulating basal PAP and in modulating the hypoxia-induced pulmonary vasoconstrictor response in immature animals. Moreover, eNOS appears to undergo up-regulation as a consequence of chronic hypoxia. PMID- 15821439 TI - Sustained cardioprotection afforded by A2A adenosine receptor stimulation after 72 hours of myocardial reperfusion. AB - This study was designed to determine whether cardioprotection afforded by A2A adenosine receptor stimulation can be sustained and to determine the effect of an A2A adenosine receptor agonist on Akt and cAMP response element binding protein (CREB) activation, as well as Hsp27 and Hsp70 protein expression in such events. The left anterior descending coronary artery was occluded for 40 minutes in anesthetized rats followed by 72 hours of reperfusion. A2A agonist (CGS21680 at 0.2 microg/kg/min) was administered for 120 minutes, starting either 5 minutes before (early) or after (late) the beginning of reperfusion. Infarct size was reduced significantly in the early compared with the control group (35.2 +/- 1.9% and 52.5 +/- 3.4%, respectively; P < 0.05), whereas no difference was observed with the late group (44.5 +/- 7.1%). After 72 hours of reperfusion, drug administration was accompanied by Akt activation (early, 121.8 +/- 17.6%; late, 118.1 +/- 16.4%; P < 0.05), as well as elevated Hsp27 expression (early, 197.2 +/ 27.7%; late, 203.8 +/- 36.8%; P < 0.05); CREB activation and Hsp70 expression were not altered. In another set of experiments in which reperfusion was limited to 15 minutes, Akt was activated only in the early group (121.8 +/- 17.6%; P < 0.05). Moreover, CREB was activated in both the early and late groups (98.4 +/- 8.3% and 107.0 +/- 6.5%, respectively; P < 0.05), whereas Hsp27 and Hsp70 expression were not altered. These results demonstrate that A2A adenosine receptor activation induces a sustained cardioprotection only if the therapy is instituted before reperfusion. This myocardial protection is associated by an early prosurvival Akt activation. CREB activation and Hsp27 content do not seem to be associated with cardioprotection because they are enhanced in both treated groups, suggesting indirect A2A agonist and pathology-related effects. PMID- 15821440 TI - Mediation of the effect of nicotine on Kir6.1 channels by superoxide anion production. AB - KATP channels are a complex of regulatory sulfonylurea receptor subunits and the pore-forming inward rectifiers such as Kir6.1. Using the whole-cell patch-clamp technique, we investigated the interaction of nicotine with the Kir6.1 subunit as well as the underlying mechanism. Stable expression of Kir6.1 in HEK-293 cells yielded a detectable inward rectifier KATP current. This inward current was significantly inhibited by PNU-37883A and by a specific anti-Kir6.1 antibody. Nicotine at 30 and 100 microM increased Kir6.1 currents by 42 +/- 11.8% and 26.2 +/- 14.6%, respectively (n = 4-6, P < 0.05). In contrast, nicotine at 1-3 mM inhibited Kir6.1 currents (P < 0.05). Nicotine at 100 microM increased the production of superoxide anion (O2) by 20.3 +/- 5.7%, whereas at 1 mM it significantly decreased the production of O2 by 37.7 +/- 4.3%. Coapplication of hypoxanthine (HX) and xanthine oxidase (XO) to the transfected HEK-293 cells resulted in a significant and reproducible increase in Kir6.1 currents (P < 0.05). The stimulatory effect of HX/XO on Kir6.1 current was abolished by tempol, a scavenger of O2. Tempol also abolished the stimulatory effect of 30 muM nicotine on Kir6.1 currents. In conclusion, nicotine stimulates Kir6.1 channel at least in part through the production of O2. PMID- 15821441 TI - Effects of KATP channel modulation on myocardial glycogen content, lactate, and amino acids in nonischemic and ischemic rat hearts. AB - ATP-sensitive potassium (KATP) channels are involved in the mechanisms underlying ischemic preconditioning. KATP channels open during ischemia, presumably secondary to intracellular metabolic alterations. The direct effects of KATP channel modulation on myocardial metabolism have not been studied. The aim of the present study was to investigate whether a KATP opener (diazoxide) and blocker (glibenclamide) modulates myocardial glycogen, lactate, and amino acid content before, during, and after ischemia. In isolated perfused rat hearts, we investigated the effect of diazoxide (30 microM) and glibenclamide (10 microM) administered 15 minutes before ischemia on myocardial glycogen, lactate, and amino acid content before, during, and after ischemia. Diazoxide increased left ventricular developed pressure during reperfusion (P < 0.05) and decreased myocardial glycogen depletion (P < 0.05) and lactate accumulation (P < 0.05) during ischemia compared with the control group. Glibenclamide decreased myocardial glycogen content (P < 0.05) and increased myocardial lactate (P < 0.05) and alanine (P < 0.05) content before ischemia and reduced myocardial glycogen content after ischemia (P < 0.05) compared with control. KATP channel activation by diazoxide modulates myocardial metabolism. These findings suggest that activation of KATP channels protects against ischemia-reperfusion injury by a mechanism that involves decreased energy depletion. PMID- 15821442 TI - Alpha v integrin affinity/specificity and antiangiogenesis effect of a novel tetraaza cyclic peptide derivative, SU015, in various species. AB - The present study was undertaken to define the alpha v beta3 and alpha v beta5 binding potency and specificity of SU015, an integrin antagonist. SU015 inhibited alpha v beta3-mediated human umbilical vein endothelial cell or 293/beta3 transfected CHO cell adhesion to fibrinogen, with IC50 values of 0.21 +/- 0.11 muM and 0.32 +/- 0.02 microM. SU015 demonstrated comparable affinity to alpha v beta5 as compared with alpha v beta3 affinity, as well as a relatively high degree of specificity for human alpha v beta3- and alpha v beta5-mediated functions, as compared with other human integrins, including alphaIIbbeta3 (IC50 >100 microM), alpha5/beta1 (IC50 >100 microM), and alpha4/beta1 (IC50 >100 microM). SU015 demonstrated different degrees of species specificity in blocking alpha v beta3-mediated cellular adhesion, with relatively higher affinity to monkey (IC50 = 0.10 microM) and dog (IC50 = 1.30 microM) endothelial or smooth muscle cell alpha v beta3-mediated adhesion. Additionally, SU015 demonstrated a high degree of alpha v beta3 and alpha v beta5 specificity as compared with alpha4beta1-, alpha5beta1-, or alpha IIb beta3-mediated binding in the above species. In conclusion, SU015 is an alpha v beta3 and alpha v beta5 antagonist with relatively higher potency and specificity as compared with alpha IIb beta3, alpha5beta1, or alpha4beta1 integrins. Additionally, comparable alpha v beta3 and alpha v beta5 affinity for SU015 was demonstrated with human and monkey endothelial cells. These data also suggest that this bicyclic RGD analogue linked to a linker at the bottom leaves the RGD at the top available for binding and allows for conjugation with radioisotope for imaging and radiotherapy. PMID- 15821443 TI - HMG-CoA reductase inhibitors reduce I kappa B kinase activity induced by oxidative stress in monocytes and vascular smooth muscle cells. AB - Reactive oxygen species, such as superoxide anion (O2-) and hydrogen peroxide (H2O2), may act as second messengers of intracellular signaling and play a key role in the pathogenesis of atherosclerosis. The nuclear factor kappaB (NF-kappa B) is a redox-sensitive transcription factor that is involved in this process. The aim of the present study was to investigate the molecular mechanisms of action of statins on cultured vascular smooth muscle cells (VSMC) and monocytic cells (THP-1) under oxidative stress. In THP-1 and cultured VSMC, O2- caused an increase in NF-kappa B activation (P < 0.05) that was correlated with inhibitory I kappa B-alpha degradation. Atorvastatin or simvastatin decreased NF-kappa B activation induced by oxidative stress by around 50% in both cell types and was correlated with the I kappa B-alpha levels. In monocytes, O2- increased I kappa B kinase (IKK)-1 and IKK-2 activity (P < 0.05) and p38 and p42/44 activation and phosphorylation, which was reduced by statins. PD 98059 (p42/44 inhibitor) and SB20358 (p38 inhibitor) decreased NF-kappa B binding activity and prevented I kappa B-alpha degradation. However, we only observed a reduction in IKK-1 and IKK 2 activity with PD98059. Statins diminish NF-kappa B activation elicited by oxidative stress through the inhibition of IKK-1/-2, p38, and p42/44 activation. These data may help to further understand the molecular mechanisms of statins in cardiovascular disease. PMID- 15821444 TI - Triptans induce vasoconstriction of human arteries and veins from the thoracic wall. AB - A common side effect of migraine treatment with triptans is chest symptoms. The origin of these symptoms is not known. The aim of the present study was to examine the vasocontractile effect of triptans in human arteries and veins from the thoracic wall and in coronary artery bypass grafts. In vitro pharmacology experiments showed that the 5-hydroxytryptamine (5-HT) type 1B and 1D receptor agonists, eletriptan, naratriptan, rizatriptan, sumatriptan, and zolmitriptan, induced vasoconstriction in the thoracic blood vessels from 38% to 57% of the patients. 5-carboxamidotryptamine (5-CT) and sumatriptan elicited a vasoconstriction that was antagonized by the 5-HT1B receptor antagonist SB224289, whereas the 5-HT1D receptor antagonist BRL115572 had no effect. 5-HT induced a contraction that was inhibited by the 5-HT2A receptor antagonist ketanserin. 5 HT2A, 5-HT1B, and 5-HT1D receptor mRNA levels were detected by real-time PCR in all blood vessels studied. In conclusion, triptans induce vasoconstriction in arteries and veins from the thoracic wall, most likely by activation of 5-HT1B receptors. This response could be observed in only 38% to 57% of the patients, which may provide an explanation for why a similar number of patients experience chest symptoms as a side effect of migraine treatment with triptans. PMID- 15821445 TI - Influence of diabetes mellitus, hypercholesterolemia, and their combination on EDHF-mediated responses in mice. AB - The endothelium synthesizes and releases several vasodilator substances, including vasodilator prostaglandins, NO, and EDHF. NO-mediated relaxations are reduced by various risk factors, such as diabetes mellitus and hypercholesterolemia. However, it remains to be elucidated whether EDHF-mediated relaxations also are reduced by those factors and their combination. In this study, we addressed this point in mice. We used small mesenteric arteries from control, diabetic (streptozotocin-induced), apolipoprotein-E-deficient (ApoE-/-), and diabetic ApoE-/- mice. In control mice, endothelium-dependent relaxations to acetylcholine were largely mediated by EDHF. This EDHF-mediated component was slightly reduced in diabetic mice, preserved in ApoE-/- mice, and markedly reduced in diabetic ApoE-/- mice with an increase in NO-mediated component and a negative contribution of indomethacin-sensitive endothelium-derived contracting factor (EDCF). Endothelium-independent relaxations to sodium nitroprusside or NS1619, a direct opener of calcium-activated K channels, were attenuated in ApoE /- and diabetic ApoE-/- mice. Endothelium-dependent hyperpolarizations were significantly reduced in diabetic mice, preserved in ApoE-/- mice, and again markedly reduced in diabetic ApoE-/- mice. These results indicate that hypercholesterolemia alone minimally affects the EDHF-mediated relaxations, and diabetes mellitus significantly attenuated the responses, whereas their combination markedly attenuates the responses with a compensatory involvement of NO and a negative contribution of EDCF. PMID- 15821447 TI - Blood pressure variability: its measurement and significance in hypertension. AB - The occurrence of blood pressure fluctuations over time has been documented since the 18th century, but the clinical importance of this phenomenon is only now being recognized. The introduction of ambulatory blood pressure monitoring in the late 1960s represented a major step forward in the study of blood pressure behaviour and helped to characterize the relationship between blood pressure variability and cardiovascular disease. In hypertension, blood pressure variability increases with increasing blood pressure and correlates closely with target-organ damage, independently of absolute blood pressure values. This has important consequences for treatment, which in the past has focused on reducing mean blood pressure values as the main goal. Experimental evidence suggests that drugs capable of buffering or reducing blood pressure variability may confer additional benefits on target-organ protection. Effective target-organ protection could best be afforded by antihypertensive agents that provide efficient 24-h blood pressure control and also stabilize blood pressure variability. Mathematical indices, such as the trough:peak ratio and the smoothness index, provide useful measures of the homogeneity of the antihypertensive effect over 24 h; optimum control is provided by drugs with a trough:peak ratio close to 1 and a smoothness index > 1, as is observed with long-acting drugs such as telmisartan or amlodipine. Recently, a direct relationship was demonstrated between homogeneous blood pressure control and treatment-induced regression of left ventricular hypertrophy, emphasizing the importance of smooth 24-h blood pressure control. In conclusion, the goals of antihypertensive treatment should consider the reduction of both 24-h mean blood pressure and its variability. Long-lasting drugs or drug combinations are preferable to ensure a homogeneous and smooth 24-h blood pressure profile. PMID- 15821448 TI - Left ventricular hypertrophy as a predictor of cardiovascular risk. AB - Left ventricular hypertrophy (LVH) is a potent, independent predictor of cardiovascular events, particularly in hypertension, in which it dramatically increases the risk of stroke, coronary heart disease and heart failure. LVH is predominantly a surrogate marker for the effects of other risk factors integrated over time, but it may also contribute directly to cardiovascular disease through pathological changes in cardiac structure. The influence of blood pressure is central to LVH pathology, with 24-h blood pressure being more predictive of LVH than single clinic measurements. Blood pressure variation throughout the day is also emerging as an important correlate of LVH, and a strong association has been found between the early morning blood pressure rise and increased left ventricular mass. Antihypertensive treatment can reverse LVH, and preliminary studies suggest that this improves cardiovascular outcome and long-term prognosis. Most classes of antihypertensive agent show some effect on LVH regression, with the notable exceptions of minoxidil and hydralazine. However, many of the data regarding LVH regression come from small, poor-quality trials or from meta-analyses of these studies. In the few well-conducted studies that are available, certain classes of antihypertensive drugs are more effective than others. Those that target angiotensin II, such as the angiotensin II receptor blockers, appear to have a specific action on LVH that is independent of blood pressure reduction. Further high-quality studies are needed to define how LVH predicts cardiovascular risk, which agents are most effective at eliciting LVH regression and how such reversal can affect cardiovascular outcome. PMID- 15821449 TI - Lowering blood pressure for cardiovascular risk reduction. AB - The association between blood pressure and cardiovascular risk is continuous, with no known lower threshold; epidemiologically, the lower the blood pressure, the better. In reality, the relationship must be J- or U-shaped, as a blood pressure of zero is associated with 100% mortality. However, the level of blood pressure control below which risk increases is well below that achieved in clinical practice, and reducing blood pressure is a cornerstone of strategies to reduce cardiovascular risk. Even relatively small reductions in blood pressure (systolic blood pressure 10-12 mmHg, diastolic blood pressure 5-6 mmHg) substantially reduce cardiovascular risk. Optimal cardiovascular protection is achievable through early and aggressive blood pressure control, but precisely which agents confer the greatest benefits for cardiovascular protection remains widely debated. Angiotensin II receptor blockers (ARBs) appear to be unique in providing additional protection beyond blood pressure control, whereas similar claims for other agents do not withstand close scrutiny. Nearly all patients with hypertension require several antihypertensive treatments to reach their target blood pressure, and it is important to choose treatments that are well tolerated and have complementary modes of action. For this reason, ARBs such as telmisartan emerge as logical choices for combination therapy, particularly when combined with a diuretic, as they fulfil all the essential requirements for combination therapy and are effective in a wide range of different types of patient. Regrettably, a rigorous approach to blood pressure control using multiple agents is still rare in general practice, partly because of a lack of understanding and partly because of 'professional non-compliance'. Prescribing habits must change, and soon, if we are to avoid an upsurge in cardiovascular complications. PMID- 15821450 TI - Relevance of blood pressure variation in the circadian onset of cardiovascular events. AB - Ambulatory blood pressure monitoring enables the recording of the circadian rhythm of blood pressure under everyday circumstances, with the majority of individuals displaying diurnal variations in both systolic and diastolic blood pressures. During sleep, blood pressure in most people is between 10% and 20% lower than the mean daytime value. On arousal and the start of day-to-day activities, there is a surge in blood pressure that may last for between 4 and 6 h. Extensive evidence shows that ambulatory blood pressure is superior to office values in predicting cardiovascular risk. Cardiovascular events, such as myocardial infarction, ischaemia and stroke are more frequent in the morning hours, soon after waking, than at other times of day. Circadian variations in biochemical and physiological parameters help to explain the link between acute cardiovascular events and the early morning blood pressure surge. Recent observations in elderly Japanese individuals demonstrate that greater early morning blood pressure surges are related to an increased incidence of overt cerebrovascular disease; individuals with the greatest increases in blood pressure on awakening also had the greatest prevalence of silent ischaemic events and were more likely to experience multiple infarcts. Antihypertensive drugs that provide blood pressure control at the time of the early morning surge should provide greater protection against target-organ damage and enhance patient prognosis. Ambulatory blood pressure monitoring may be particularly helpful in assessing the circadian pharmacodynamics of such antihypertensive drugs. The technique has demonstrated, for example, a significantly greater reduction in blood pressure for the last 6 h of the 24-h dosing interval with telmisartan compared with valsartan. PMID- 15821451 TI - Managing the patient at risk for a second stroke. AB - Cerebrovascular disease is a major cause of mortality world-wide, and the prevalence is expected to increase as a result of projected demographic trends. Aggressive antihypertensive therapy is one intervention that has proven highly effective in reducing the risk of stroke, with relatively small blood pressure reductions affording measurable benefit even in patients not conventionally considered hypertensive. Comparative clinical trials are revealing evidence of differential impacts of antihypertensive classes on the incidence of cerebrovascular disease that will probably be important for therapeutic choice in patients with risk factors for stroke. In particular, the role of the renin angiotensin system in cerebrovascular disease has come under scrutiny as a result of evidence that angiotensin II receptor blockers (ARBs), but perhaps not angiotensin converting enzyme inhibitors, can reduce the risk of a first stroke to a greater degree than might be expected from their effects on blood pressure alone. Although preclinical evidence suggests that there are differential effects of the type 1 and type 2 receptor activation, the clinical relevance of this is not yet known. Furthermore, the effect on the incidence of stroke conferred by blood pressure control in the early morning hours - the time when the incidence of strokes peaks--has not been tested. Some evidence for the beneficial effect of an ARB on secondary stroke prevention comes from the MOrbidity and mortality after Stroke --Eprosartan compared with nitrendipine in Secondary prevention study (MOSES), which showed that the ARB protected against cerebro- and cardiovascular events in hypertensive patients with a previous stroke over and above the protection offered by blood pressure control. These hypotheses are among those being examined in two current large-scale trials: the Prevention Regimen For Effectively avoiding Second Strokes (PRoFESS), and The ONgoing Telmisartan Alone and in combination with Ramipril Global Endpoint Trial (ONTARGET) Trial Programme. PMID- 15821452 TI - The cardiovascular continuum and renin-angiotensin-aldosterone system blockade. AB - A progressive chain of pathophysiological events links cardiovascular risk factors to clinical manifestations of disease and life-threatening cardiovascular events. This chain--the cardiovascular continuum--underlies cardiovascular disease and holds the key to its prevention and treatment. Progressive tissue damage can result in morbidity from congestive heart failure, end-stage heart disease, nephrotic proteinuria and dementia and, eventually, death from cardio- or cerebrovascular causes. The renin-angiotensin-aldosterone system (RAAS) is involved at all stages of the cardiovascular continuum, because the effector molecules of the RAAS, angiotensin II in particular, have direct pathobiological effects on a variety of tissues, including the endothelium, vascular smooth muscle and the renal mesangium. Clinical trials of angiotensin II receptor blockers (ARBs) and angiotensin-converting enzyme (ACE) inhibitors have demonstrated the essential validity of this hypothesis. Interruption of the RAAS has been shown to reduce cardiovascular morbidity and mortality in patients with left ventricular hypertrophy, heart failure and post-myocardial infarction, as well as renal disease in patients with type 2 diabetes. Key questions remain, however. What are the clinical effects of combination ARB and ACE inhibitor treatment? How will combinations of RAAS blockade with other agents, such as statins, affect the cardiovascular continuum? Answers to these questions will require well-planned, adequately powered clinical trials, such as the Programme of Research tO evaluate Telmisartan End-organ proteCTION (PROTECTION) and the ONgoing Telmisartan Alone and in combination with Ramipril Global Endpoint Trial (ONTARGET) programmes. However, it is already clear that RAAS blockade is an essential part of blocking progression along the cardiovascular continuum. PMID- 15821456 TI - Is it time to change the training of urology residents in the United States? PMID- 15821457 TI - Rethinking urolithiasis. PMID- 15821458 TI - Urinary incontinence: a diverse condition. PMID- 15821459 TI - A problem with a chaperone. PMID- 15821460 TI - Limitations in the use of serum prostate specific antigen levels to monitor patients after treatment for prostate cancer. AB - PURPOSE: We reviewed the literature to help clarify the benefits and/or hazards associated with monitoring serum prostate specific antigen (PSA) after treatment with surgery or radiation therapy (RT) for nonmetastatic prostate cancer. MATERIALS AND METHODS: A search was performed for 1990 to 2004 using the MEDLINE database, CancerLit database and reference lists of relevant studies to obtain articles addressing the use of serum PSA to follow patients after treatment for prostate cancer. Studies were reviewed to determine 1) if serial PSA monitoring provides an early and accurate surrogate assessment of cancer cure or treatment failure, 2) if any pattern in the PSA profile after treatment provides conclusive evidence of early local vs systemic failure, 3) the magnitude of the lead time to clinical failure that serial PSA monitoring may provide and 4) if the early identification of biochemical failure (BF) with earlier intervention improves outcome. RESULTS: Although a lower PSA nadir after treatment with RT has been associated with cancer cure, 5% to 25% of patients ultimately have failure (beyond 5 years) even with the most optimal biochemical response. The most appropriate BF definitions to use after treatment for prostate cancer with RT remains controversial due to substantial differences in their accuracy, sensitivity, specificity and positive predictive value for clinical outcome. No pattern of PSA kinetics after treatment has conclusively been associated with a specific recurrence site. Biochemical failure definitions in patients treated with RT appear to provide a 6 to 18 month lead time to clinical failure but there are only limited published data to suggest that early intervention of any type (androgen deprivation, RT, surgery, etc) impacts survival. CONCLUSIONS: The overall benefit of monitoring serum PSA after treatment for prostate cancer remains controversial. Considering the potential dangers associated with incorrectly assuming the efficacy of new forms of treatment, the toxicity of administering salvage therapies of uncertain efficacy after BF has been identified and the anxiety associated with tracking posttreatment serum PSA, additional studies must be done to determine the appropriate use of this marker in properly treating patients after therapy. PMID- 15821461 TI - Mechanics of wound healing and importance of Vacuum Assisted Closure in urology. AB - PURPOSE: We discuss the mechanisms of wound healing and our experience with the Vacuum Assisted Closure device (Kinetic Concepts, Inc., San Antonio, Texas) for complex urogenital wounds. MATERIALS AND METHODS: The literature obtained from a Medline search on wound healing, wound failure and vacuum assisted closure was reviewed. In addition, we reviewed our experience with negative pressure wound therapy. RESULTS: Wound healing is a complex interaction between the reticuloendothelial and immune systems, in addition to correctable internal and external factors. Understanding the healing process improves outcomes and decreases patient morbidity. Negative pressure wound therapy has hastened wound healing and it adds significant improvement in the arsenal of choices available. CONCLUSIONS: Vacuum Assisted Closure is a therapeutic alternative that complements surgical and medical intervention in patients with complex wounds. PMID- 15821462 TI - Congenital unilateral hydronephrosis: a review of the impact of diuretic renography on clinical treatment. AB - PURPOSE: The natural history of and optimal treatment for asymptomatic congenital hydronephrosis is unknown, mainly because most published series are nonrandomized or retrospective with nonstandardized followup regimens. In this survey we provide an overview of the different strategies used for the treatment of congenital unilateral hydronephrosis. MATERIALS AND METHODS: A comprehensive literature review of clinical studies of asymptomatic congenital unilateral hydronephrosis is presented. RESULTS: Different criteria for surgical intervention and different followup regimens have been used. Identification of the population at risk has been attempted but results are ambiguous. Surgery is done in approximately 25% of cases when using deteriorating differential function, symptoms and/or increasing hydronephrosis as criteria for operation. A nonstandardized renographic assessment does not compromise evaluation of the ideal followup but it may compromise the functional outcome. CONCLUSIONS: More studies of kidneys with decreased function, severe hydronephrosis or poor drainage investigated with a standardized protocol are needed to clarify the natural history and establish optimal guidelines for treatment and followup. PMID- 15821463 TI - Fibrin glue-oxidized cellulose sandwich for laparoscopic wedge resection of small renal lesions. AB - PURPOSE: We present our experience with standard laparoscopic wedge resection of small renal tumors using a fibrin glue-oxidized cellulose sandwich for hemostasis. MATERIALS AND METHODS: From May 2002 to December 2003, 15 patients underwent laparoscopic wedge resection for a total of 15 nonhilar renal masses. Renal hilar clamping was performed in 1 patient and no sutures were placed. We used ultrasonic shears and an argon beam coagulator to resect and then coagulate the tumor bed. Tisseel (Baxter Corp., Deerfield, Illinois) was applied to the resection bed, followed by a layer of oxidized, regenerated cellulose (Surgicel, Ethicon, Somerville, New Jersey) and a final layer of Tisseel. RESULTS: Mean preoperative tumor size was 2.2 x 2.2 x 2.1 cm. Lesions were subdivided based on the percent of the lesion that extended beyond the renal parenchymal border on computerized tomography as exophytic-greater than 60% in 6 cases, endophytic-less than 40% in 4 and mesophytic-40% to 60% in 5. Mean operative time was 3.8 hours (range 3 to 5). Mean blood loss was 108 ml (range 20 to 300). No patient required blood transfusion. There was no significant difference in blood loss or change in creatinine among the endophytic, exophytic and mesophytic groups (150, 121 and 93 ml, and 0.03, 0.07 and 0.04 mg/dl, respectively). Margin status was negative in all cases (mean thickness 3.2 mm). Average hospital stay and time to the resumption of oral intake were 2.7 and 1.4 days, respectively. CONCLUSIONS: Small exophytic or mesophytic renal lesions can be safely excised laparoscopically without vessel clamping. Excellent hemostasis was achieved in each case with the newer hemostatic agents. PMID- 15821465 TI - Evaluation of the intact specimen after laparoscopic radical nephrectomy for clinically localized renal cell carcinoma identifies a subset of patients at increased risk for recurrence. AB - PURPOSE: Laparoscopic radical nephrectomy (LRN) is emerging as a standard approach for low stage renal cell carcinoma (RCC). Some suggest that specimen morcellation for extraction results in less morbidity and a faster recovery. However, morcellation may preclude accurate pathological staging and may hinder precise pathological grading. With pathological evaluation of an intact specimen we identified patients harboring high risk disease that was not anticipated preoperatively, defined as pT2 high grade (G3-4) or lesions greater than pT2. MATERIALS AND METHODS: We retrospectively reviewed the records of 192 patients who underwent LRN for renal lesions at The University of Texas M. D. Anderson Cancer Center between April 2002 and April 2004. RESULTS: A total of 192 patients underwent LRN for presumed RCC. In all cases specimens were removed intact. Of these cases 137 were cT1/T2 N0 M0 and had a final pathological diagnosis of RCC. All surgical margins were negative. Of the 137 patients 40 (29.2%) were at increased risk for recurrence based on high risk features. Specifically up staging from cT1-2 to pT3 disease occurred in 30 patients (21.9%) while 12 patients (8.8%) were found to have pT2 G3-4 disease. All 40 patients with high risk disease underwent more intensive surveillance and 17 (43%) participated in adjuvant systemic therapy trials. CONCLUSIONS: In our study 21.9% of patients with clinical T1-2 disease were pathologically up staged and 29.2% were identified as being at high risk for recurrence after intact specimen extraction for localized RCC. These patients are candidates for more intensive followup treatment and may benefit from enrollment in adjuvant therapy protocols. PMID- 15821464 TI - Evaluation and management of renal tumors in the Birt-Hogg-Dube syndrome. AB - PURPOSE: Herein we describe the evaluation and management of renal tumors in Birt Hogg-Dube (BHD), an autosomal dominant disorder predisposing to cutaneous fibrofolliculomas, pulmonary cysts, spontaneous pneumothorax and renal tumors. MATERIALS AND METHODS: A total of 124 affected individuals underwent comprehensive clinical evaluation, including body computerized tomography, to determine cutaneous, pulmonary and renal manifestations of BHD. Of these individuals 14 had their renal tumors managed at our institution. RESULTS: Of the 124 BHD affected individuals 34 (27%) had renal tumors of various histologies, most commonly hybrid oncocytic tumor and chromophobe renal carcinoma. Average age at renal tumor detection was 50.4 years and multiple tumors were found in a majority of patients. Some patients with renal tumors were identified that did not have the characteristic cutaneous hallmarks of BHD. In 4 of the 14 patients treated at our institution small (less than 3 cm) renal tumors were observed, while 10 others underwent a total of 12 renal procedures, including 4 radical and 8 partial nephrectomies. At a median of 38 months of followup 5 of these 10 patients remained free of disease, 3 had small renal tumors and 2 died of metastatic renal cancer. CONCLUSIONS: Patients with BHD are at risk for multiple renal tumors that are often malignant and can metastasize. Individuals at risk or affected by BHD should be radiographically screened for renal tumors at periodic intervals and they are best treated with nephron sparing surgical approaches. Genetic testing for this syndrome is now available. PMID- 15821466 TI - pT1 substaging in renal cell carcinoma: validation of the 2002 TNM staging modification of malignant renal epithelial tumors. AB - PURPOSE: Tumor size has been used as one of the criteria to stratify renal cell carcinoma (RCC) into different pathological stages (pT). The recent 2002 UICC/TNM classification of malignant epithelial renal tumors is modified to substratify pT1 RCC into pT1a (less than 4.0 cm) and pT1b (greater than 4.0 but less than 7.0 cm). In this study we ascertained if this stage modification has prognostic relevance. MATERIALS AND METHODS: A total of 259 consecutive radical nephrectomy specimens of organ confined RCC from 1970 to 1997 at 1 institution, including 153 of conventional RCC (CRCC), 71 of papillary RCC, 28 of chromophobe RCC, 1 of collecting duct carcinoma and 6 of RCC not otherwise specified, with a mean clinical followup of 7.5 years (median 6.4) were included in the study. RESULTS: There were 115 pT1a (44.4%), 95 pT1b (36.7%) and 49 pT2 tumors (18.9%). Disease recurrences (DR) and disease specific death occurred in 2 (1.7%) and 0 cases (0%) of pT1a, 7 (7.3%) and 5 (5.3%) of pT1b, and 16 (32.6%) and 12 (24.5%) of pT2. DR for pT1b was higher compared with pT1a (all histological subtypes RR 3.68), although this difference was not statistically significant (p = 0.106). If only CRCCs were analyzed, DR in the pT1b group was statistically higher compared with pT1a (RR 8.54, p = 0.047). Disease specific survival in pT1a could not be evaluated because no deaths occurred in this subgroup. DR and disease specific survival were significantly different between pT1b and pT2 tumors for all histological subtypes (RR 5.51, p = 0.001 and 5.49, p = 0.001) and for the CRCC subtype (RR 5.50, p = 0.001 and 5.18, p = 0.005, respectively). Using size as a continuous variable the logarithmic change in tumor size was a significant predictor of DR (RR 8.82, p = 0.001). All statistical analyses were adjusted for age and sex. CONCLUSIONS: Substaging RCC into pT1a and pT1b yields prognostically important information, validating the 2002 TNM modification for malignant renal epithelial malignancies. The substratification of pT1 is particularly useful in tumors with CRCC histology. PMID- 15821467 TI - Using tumor markers to predict the survival of patients with metastatic renal cell carcinoma. AB - PURPOSE: Approximately 30% of renal cell carcinomas (RCCs) present as metastatic disease. Molecular markers have the potential to characterize accurately the biological behavior of tumors and they may be useful for determining prognosis. MATERIALS AND METHODS: A custom tissue array was constructed using clear cell RCC from 150 patients with metastatic RCC who underwent nephrectomy prior to immunotherapy. The tissue array was stained for 8 molecular markers, namely Ki67, p53, gelsolin, carbonic anhydrase (CA)9, CA12, PTEN (phosphatase and tensin homologue deleted on chromosome 10), epithelial cell adhesion molecule and vimentin. Marker status and established clinical predictors of prognosis were considered when developing a prognostic model for disease specific survival. RESULTS: On univariate Cox regression analysis certain markers were statistically significant predictors of survival, namely CA9 (p <0.00001), p53 (p = 0.0072), gelsolin (p = 0.030), Ki67 (p = 0.036) and CA12 (p = 0.043). On multivariate Cox regression analysis that included all markers and clinical variables CA9 (p = 0.00002), PTEN (p <0.0001), vimentin (p = 0.0032), p53 (p = 0.028), T category (p = 0.0025) and performance status (p = 0.0013) were significant independent predictors of disease specific survival and they were used to construct a combined molecular and clinical prognostic model. The bias corrected concordance index (C-index) of this combined prognostic model was C = 0.68, which was significantly higher (p = 0.0033) than that of a multivariate clinical predictor model (C = 0.62) based on the UCLA Integrated Staging System (T category, histological grade and performance status). CONCLUSIONS: In patients with clear cell RCC a prognostic model for survival that includes molecular and clinical predictors is significantly more accurate than a standard clinical model using the combination of stage, histological grade and performance status. PMID- 15821468 TI - Serum carotenoids and other antioxidative substances associated with urothelial cancer risk in a nested case-control study in Japanese men. AB - PURPOSE: We assayed whether high serum carotenoids and antioxidative substances decrease the risk of urothelial cancer in a case-control study nested in a community based cohort in Japan, that is the Japan Collaborative Cohort Study. MATERIALS AND METHODS: Information on subject life-styles and serum were collected in 1988 to 1990 and subjects were followed as late as 1999. Individuals who had or died of urothelial cancer and controls were matched for study area, sex and age. Serum was stored at -80C and analyzed in 2003. Of 14,097 male and 25,662 female subjects 40 to 79 years old there were 42 cases, which were matched to 124 controls. RESULTS: The OR for the highest to lowest tertile of serum concentration was 0.28 (95% CI 0.07 to 1.15, trend p = 0.08) for beta-carotene, 0.36 (95% CI 0.10 to 1.27, trend p = 0.10) for total carotenes and 0.31 (95% CI 0.09 to -1.09, trend p = 0.09) for total carotenoids after adjustment for smoking in addition to matching variables. High concentrations of tocopherols and xanthophylls slightly tended to decrease the risk of urothelial cancer. In contrast, serum retinol, oxidized low density lipoprotein and Cu/Zn-superoxide dismutase were not associated with urothelial cancer risk. CONCLUSIONS: Our results suggest that high serum carotenoids may decrease the risk of urothelial cancer with carotenes more effective than xanthophylls. PMID- 15821469 TI - Bacillus calmette-guerin Tokyo172 substrain for superficial bladder cancer: characterization and antitumor effect. AB - PURPOSE: We investigated the preparation of bacillus Calmette-Guerin (BCG) Tokyo172 substrain (Japan BCG Laboratory, Ltd., Tokyo, Japan) on the characteristics of bacilli and antitumor activity in a mouse model in comparison with a preparation of the Connaught substrain (Aventis Pasteur, Ltd., Toronto, Ontario, Canada). MATERIALS AND METHODS: Lyophilized BCG preparations of Tokyo172 and Connaught for superficial bladder cancer were tested. The number of bacilli and cfu per dose, dispersion, size and attachment to murine bladder tumor cells were determined after reconstitution. Antitumor activity was assessed by intradermal injection of tumor cells with various doses of either BCG preparation into the flanks of syngeneic mice, followed by the observation of tumor suppression and survival in mice. RESULTS: Each dose of Tokyo172 had about half the bacilli in a dose of Connaught but the cfu content was about 13-fold higher for Tokyo172 than for Connaught. After reconstitution Tokyo172 bacilli were better dispersed with fewer aggregates than Connaught bacilli. Tokyo172 bacilli were about half as long as Connaught bacilli and Tokyo172 bacilli showed better attachment to tumor cells in vitro. In mice Tokyo172 achieved similar tumor suppression at a lower dose than Connaught. CONCLUSIONS: High viability, good dispersion and efficient binding to tumor cells by BCG bacilli in the Tokyo172 preparation seem to be the main reasons for the lower clinical dose of this preparation compared with the Connaught preparation (18 vs 81 mg dry weight). PMID- 15821470 TI - Does body mass index affect survival of patients undergoing radical or partial cystectomy for bladder cancer? AB - PURPOSE: Obesity is estimated to account for up to 20% of all cancer deaths. We evaluated the effect of body mass index (BMI) on survival in patients undergoing radical or partial cystectomy for bladder cancer. MATERIALS AND METHODS: Pathological and medical records on 300 consecutive patients undergoing radical or partial cystectomy for invasive bladder cancer between January 1990 and December 1993 were reviewed. The standard WHO definition of BMI was used, that is normal weight-less than 25 kg/m, overweight-25 to 29.9 kg/m and obesity-30 kg/m or greater. Sufficient data were available on 288 of 300 patients (96%) with cystectomy (radical in 264 and partial in 24) for analysis. RESULTS: The BMI distribution was normal weight in 34% of patients, overweight in 41% and obesity in 25%. Mean followup was 53.4 months (median 39, range 1 to 168). Median overall survival was 43.0 months (95% CI 37.1 to 58.4), while median disease specific survival was 82.5 months (95% CI 50.0 to 127.5). Multivariate analysis revealed that age greater than 65 years, pathological stage, smoking history and soft tissue margin status as significant factors impacting overall survival (p <0.05). Pathological stage was organ confined (less than pT3a) in 51% of cases. BMI was not associated with disease specific survival as a continuous (p = 0.17) or categorical (p = 0.51) variable. Although it was insignificant, unadjusted analysis showed lower disease specific mortality in patients with a BMI of less than 25 mg/kg and organ confined disease (p = 0.08). CONCLUSIONS: There was no significant association between BMI and overall or disease specific survival, although there may be a trend toward better disease specific survival in normal weight (BMI less than 25 kg/m) patients with organ confined disease (p = 0.08). PMID- 15821471 TI - Nomograms including nuclear matrix protein 22 for prediction of disease recurrence and progression in patients with Ta, T1 or CIS transitional cell carcinoma of the bladder. AB - PURPOSE: We developed and validated nomograms that accurately predict disease recurrence and progression in patients with Ta, T1, or CIS transitional cell carcinoma (TCC) of the bladder using a large international cohort. METHODS: Univariate and multivariate logistic regression models targeted histologically confirmed disease recurrence, and focused on 2,542 patients with bladder TCC from 10 participating centers. Variables consisted of pre-cystoscopy voided urine Nuclear Matrix Protein 22 (NMP22) assay, urine cytology, age and gender. Resulting nomograms were internally validated with bootstrapping. Nomogram performance was explored graphically with Loess smoothing plots. RESULTS: Overall 957 patients had recurrent TCC. Tumor grade and stage was available for 898 patients, including 24% grade I, 43% grade II, and 33% grade III; 45% stage Ta, 32% T1 and/or CIS, and 23% T2 or greater. Bootstrap corrected predictive accuracy for any TCC recurrence was 0.842; grade III Ta/T1 or CIS was 0.869; and T2 or higher stage TCC of any grade was 0.858. Virtually perfect performance characteristics were observed for the nomograms predicting any TCC recurrence or grade III Ta/T1 or CIS. The nomogram predicting T2 or higher stage TCC overestimated the observed probability for predicted values greater than 45%. CONCLUSIONS: We developed and internally validated nomograms that incorporate urinary NMP22, cytology, age and gender to predict with high accuracy the probability of disease recurrence and progression in patients with Ta, T1, and/or CIS bladder TCC. These nomograms could provide a means for individualizing followup in patients with Ta, T1, CIS bladder TCC. PMID- 15821472 TI - Higher than expected association of clinical prostate and bladder cancers. AB - PURPOSE: In this study we evaluated the risk of a second malignancy of the bladder or prostate in patients with a previous diagnosis of prostate cancer (PCa) or urothelial cancer (TCC). MATERIALS AND METHODS: We retrospectively analyzed all cases of PCa and TCC diagnosed between January 1996 and June 2003. Only PCa diagnosed due to abnormal digital rectal examination or increased prostate specific antigen were included. All patients with TCC presented with hematuria or irritative voiding symptoms and the diagnoses were confirmed with a tissue diagnosis. The incidence of lung, colon and renal cancers was also analyzed. RESULTS: A total of 816 men were diagnosed with PCa and/or TCC. Of 673 men initially diagnosed with PCa 21 had TCC. Of 149 men initially diagnosed with TCC 18 had PCa. Average age at PCa and TCC diagnosis +/- SD was 68.2 +/- 7.9 and 68.2 +/- 10.4 years, respectively. The standardized incidence ratio (SIR) of TCC in patients with PCa (SIR 4.31, 95% CI 2.411 to 7.110) and of PCa in patients with TCC (SIR 3.83, 95% CI 1.911 to 6.858) was significantly increased. There was no statistical significant difference in SIR for TCC in men with or without radiotherapy. SIR for lung, renal or colon cancer was not significantly different from what was expected. CONCLUSIONS: Patients with PCa have higher incidence of bladder cancer and those with bladder cancer have a higher incidence of PCa. This study has clinical implications in the care of these patients and it may stimulate research interest that may identify common pathways of carcinogenesis. PMID- 15821481 TI - Individualization of the biopsy protocol according to the prostate gland volume for prostate cancer detection. AB - PURPOSE: In this study we assessed the relative yield of 10 core biopsy, and the whole range of alternative 8 and 6 core biopsy protocols over that of the classic sextant biopsy protocol. We determined the optimum number of cores per biopsy according to prostate volume in patients who experienced prostate biopsy for the first time. MATERIALS AND METHODS: A total of 503 men with the indications of abnormal digital rectal examination and/or serum prostate specific antigen greater than 2.5 ng/ml were included in the study. All patients underwent a 10 core biopsy protocol with an additional 1 core from each suspicious area detected by transrectal ultrasound. Prostate volume was divided into quartiles, namely 14.9 to 35, 35.1 to 50, 50.1 to 65 and 65.1 to 150 cc. The optimum number of biopsy cores was determined in patients with different prostate volumes. RESULTS: Median age was 63 years and prostate specific antigen was 7.4 ng/ml in the whole group. Of 503 patients 159 (31.6%) were positive for prostate cancer. Cancer detection rates decreased significantly from 49.6% to 20.8% as prostate volume increased in preset quartiles. Lesion biopsies revealed the lowest unique cancer detection rates for all prostate volume quartiles (0% to 3%). There was an obvious positive trend in cancer detection rates in favor of the 10 core biopsy protocol over sextant biopsies in all patient groups. Classic sextant biopsy protocol proved to be inadequate for all prostate volumes. Among sextant biopsy protocols laterally placed cores including the apex, lateral mid gland and lateral base had the best cancer detection rates (81% to 95%). The 8 core biopsy scheme consisting of the apex, mid gland, lateral mid gland and lateral base resulted in an only 1% lower detection rate (97%) than the 10 core biopsy protocol in the lowest quartile. The yield of the 10 core biopsy protocol in patients with a prostate volume of between 35.1 and 150 cc outscored that of the optimal 8 core biopsy scheme including the apex, base, lateral mid gland and lateral base with 3% to 8% differences in the cancer detection rate. CONCLUSIONS: The 10 core biopsy protocol must be used in all group of patients except patients with a prostate volume of 14.9 to 35 cc. In patients with a prostate volume of 14.9 to 35 cc the 8 core biopsy protocol consisting of the apex, mid gland, lateral mid gland and lateral base can be used since it revealed results similar to those of the 10 core biopsy protocol. The classic sextant biopsy protocol seemed inadequate for all prostate volumes. Patients with a larger prostate had lower cancer detection rates. Transrectal ultrasound directed lesion biopsies may be omitted when using 10 core biopsy protocols since the yield of these biopsies was less than 2%. PMID- 15821482 TI - Prostate specific antigen adjusted for transition zone epithelial volume: the powerful predictor for the detection of prostate cancer on repeat biopsy. AB - PURPOSE: The indications for repeat prostate biopsy for persistently increased prostate specific antigen (PSA) in men with prostate cancer never detected on previous biopsy are not clear. In this study we determined that PSA adjusted for transition zone (TZ) epithelial volume is the most powerful predictor for detecting prostate cancer on repeat biopsy. MATERIALS AND METHODS: Repeat prostate biopsies including additional TZ cores were performed in 75 men with PSA between 4.0 and 10.0 ng/ml. TZ epithelial volume was calculated by multiplying TZ volume by the percent of epithelium, which was measured by morphometric analysis using image analysis computer software. RESULTS: Prostate cancer was detected on repeat biopsy in 19 of the 75 patients. Patients with prostate cancer had a significant smaller percent area of epithelium or glandular lumen than those without cancer. In patients without prostate cancer TZ epithelial volume significantly correlated with total PSA. According to ROC analysis PSA adjusted for TZ epithelial volume had the greatest AUC for cancer detection (0.879). This parameter was able to avoid more than 90% of unnecessary repeat biopsies with 90% sensitivity. Multiple logistic regression analysis showed that PSA complex adjusted for TZ epithelial volume was the significant independent predictor of cancer. CONCLUSIONS: PSA adjusted for TZ epithelial volume is the most powerful predictor of cancer in men who have undergone previous negative prostate biopsies and in whom PSA remains between 4.0 and 10.0 ng/ml. PMID- 15821483 TI - Prostate cancer in patients with screening serum prostate specific antigen values less than 4.0 ng/dl: results from the cooperative prostate cancer tissue resource. AB - PURPOSE: Prostate cancer can occur in patients with low screening serum prostate specific antigen (PSA) values (less than 4.0 ng/ml). It is currently unclear whether these tumors are different from prostate cancer in patients with high PSA levels (greater than 4.0 ng/ml). MATERIALS AND METHODS: From the Cooperative Prostate Cancer Tissue Resource database through March 2004, 3,416 patients with screening PSA less than 16.0 ng/ml diagnosed with prostate cancer between 1993 and 2004 were stratified in groups based on screening serum PSA. These subsets were compared for race, age at diagnosis, clinical and pathological stage, Gleason score, positive surgical margins, posttreatment recurrent disease, and vital status. RESULTS: We identified 468 (14%) patients with screening PSA less than 4.0 ng/ml, 142 (4.2%) of whom had a PSA of less than 2.0 ng/ml. This group included 40 black and 376 white patients. Men with low screening PSA treated with radical prostatectomy had smaller cancers, lower Gleason scores, lower pathological tumor (T) stage and lower PSA recurrence rates than men with high PSA levels (4 ng/ml or greater). These differences held true for men who were younger than 62 years or were white, whereas older or black men had tumor characteristics and outcomes similar to those with higher PSA levels. CONCLUSIONS: Young (younger than 62 years) or white patients with screening serum PSA less than 4.0 ng/ml had smaller, lower grade tumors and lower recurrence rates than patients with PSA 4.0 ng/ml or greater. This was not true for those older than 62 years and for black men. PMID- 15821484 TI - Ex vivo expanded human Vgamma9Vdelta2+ gammadelta-T cells mediate innate antitumor activity against human prostate cancer cells in vitro. AB - PURPOSE: We have previously identified a CD2 mediated, interleukin-12 dependent signaling pathway that inhibits activation induced cell death in mitogen stimulated human gammadelta-T cells, permitting the large-scale expansion of these cells. Herein we report the innate antitumor activity of expanded human Vgamma9Vdelta2+ gammadelta-T cells against human prostate cancer cells. MATERIALS AND METHODS: Apoptosis resistant human gammadelta-T cells were expanded in vitro from cultured human peripheral blood mononuclear cells and then enriched to high purity by immunomagnetic separation. In vitro cytotoxicity of expanded gammadelta T cells was measured against human prostate cancer cell lines using standard cytotoxicity assays. RESULTS: gammadelta-T cells derived from various donors consistently showed lytic activity against the prostate cancer cell lines DU-145 and PC-3 but not LNCaP. mAbs against Vgamma9 or Vdelta2 T-cell receptor chains as well as mAb against intercellular adhesion molecule-1 (ICAM-1) or CD18, the beta subunit of ICAM-1 counter receptors, blocked gammadelta-T cell mediated killing of prostate cancer cells. gammadelta-T cells lysed prostate cancer cell lines largely through the perforin/granzyme pathway. CONCLUSIONS: Ex vivo, expanded human Vgamma9Vdelta2+ gammadelta-T cells are able innately to recognize and kill certain human prostate tumor cell lines in vitro. The recognition and killing of prostate cancer cells occurs in a gammadelta-T-cell receptor dependent manner and it also appears to involve interactions between ICAM-1 and CD18. Because apoptosis resistant human Vgamma9Vdelta2+ gammadelta-T cells can readily be expanded to large numbers (clinical scale), these findings must be considered in the context of developing adoptive immunotherapy strategies to exploit gammadelta T cell innate immune responses to prostate cancer. PMID- 15821485 TI - Treatment of patients with high risk localized prostate cancer: results from cancer of the prostate strategic urological research endeavor (CaPSURE). AB - PURPOSE: Pretreatment risk assessment models facilitate more appropriate selection of treatment for prostate cancer. However, men with high risk disease remain a challenge with significant potential for primary treatment failure. We characterize patterns of treatment for high risk prostate cancer in a community based cohort. MATERIALS AND METHODS: In the Cancer of the Prostate Strategic Urological Research Endeavor (CaPSURE) database, a longitudinal disease registry of men with prostate cancer, we identified those with nonmetastatic, high risk disease based on T stage, tumor grade and serum prostate specific antigen (PSA). Differences in primary treatment, and the use of neoadjuvant and adjuvant therapy in patients at low, intermediate and high risk were assessed. In the high risk cohort predictors of the type of primary treatment, and the use of neoadjuvant and adjuvant androgen therapy were identified. RESULTS: Of the cancers 34%, 40% and 26% were low, intermediate and high risk, respectively. Differences in primary treatment type among the 3 risk groups were statistically significant (p <0.0001) with increasing external beam radiation therapy and androgen deprivation, and decreased surgery, brachytherapy and surveillance in men with high risk cancers. In this group older age, higher PSA and nonprivate insurance were associated with decreased use of radical prostatectomy. More than half of the men at high risk receiving radiation therapy also received androgen deprivation, which was significantly higher than in the low and intermediate risk groups (p <0.0001). Factors associated with androgen deprivation in high risk disease were primary therapy, PSA, Gleason sum, T stage, body mass index, insurance status and ethnicity. PSA and Gleason sum were the primary determinants of adjuvant radiation after prostatectomy. CONCLUSIONS: Men with high risk but nonmetastatic prostate cancer are more likely to receive radiation therapy as well as androgen deprivation with the latter as primary therapy or in conjunction with local treatment. These data stress the importance of pretreatment risk stratification, education regarding appropriate combinations of local and systemic therapies, and the consideration of novel clinical trials in patients at higher risk. PMID- 15821486 TI - 12-year outcomes following permanent prostate brachytherapy in patients with clinically localized prostate cancer. AB - PURPOSE: We reviewed the outcomes in men treated with permanent prostate brachytherapy (PPB). MATERIAL AND METHODS: A total of 1,449 consecutive patients with a mean age of 68 years treated with PPB between 1992 and 2000 and mean pretreatment prostate specific antigen (PSA) 10.1 ng/ml were included in this study. Of the patients 55% presented with Gleason 6 tumors and 28% had Gleason 7 disease. A total of 400 patients (27%) were treated with neoadjuvant hormones and 301 (20%) were treated in combination with external radiation plus PPB. Several biochemical freedom from recurrence (BFR) definitions were determined. Statistical analysis consisted of log rank testing, Kaplan-Meier estimates and Cox regression analysis. RESULTS: Median followup was 82 months with 39 patients at risk at for 144 months. Overall and disease specific survival at 12 years was 81% and 93%, respectively. The 12-year BFR was 81%, 78%, 74% and 77% according to the American Society for Therapeutic Radiology and Oncology (ASTRO), ASTRO Kattan, ASTRO-Last Call and Houston definitions, respectively. The 12-year ASTRO Kattan BFR using risk stratification was 89%, 78% and 63% in patients at low, intermediate and high risk, respectively (p = 0.0001). Multivariate analysis identified the dose that 90% of the target volume received (p <0.0001), pretreatment PSA (p = 0.001), Gleason score (p = 0.002), the percent positive core biopsies (p = 0.037), clinical stage (p = 0.689), the addition of hormones (p = 0.655) and the addition of external radiation (p = 0.724) for predicting BFR ASTRO. Five-year disease specific survival was 44% in patients with a PSA doubling time of less than 12 months vs 88% in those with a PSA doubling time of 12 months or greater (p = 0.0001). CONCLUSIONS: PPB offers acceptable 12-year BFR in patients who present with clinically localized prostate cancer. Implant dosimetry continues as an important predictor for BFR, while the addition of adjuvant therapies such as hormones and external radiation are insignificant. In patients who experience biochemical failure it appears that PSA doubling time is an important predictor of survival. PMID- 15821487 TI - A prospective analysis of the time to normalization of serum androgens following 6 months of androgen deprivation therapy in patients on a randomized phase III clinical trial using limited hormonal therapy. AB - PURPOSE: Patients with prostate cancer are treated with neoadjuvant, adjuvant and intermittent therapy with gonadotropin-releasing hormone agonists (GnRH-A). While these are largely successful in decreasing testosterone (T) and dihydroxytestosterone (DHT) to castrate levels, discontinuation of such therapy often results in continued suppression of androgens for variable periods of time. We present the largest published series of patients evaluating the timing of T and DHT increase after cessation of GnRH therapy. MATERIALS AND METHODS: Serial T and DHT measurements were prospectively obtained every 3 months while on GnRH-A then monthly upon discontinuation of GnRH-A. Analysis of time from the second 3 month GnRH-A administration to T and DHT increase was undertaken. RESULTS: A total of 80 evaluable patients had a median time to T 50 ng/dl or greater of 12.9 weeks and a median time to T normalization (212 ng/dl or greater) of 16.6 weeks. Low baseline T was associated with a prolonged time to T 212 ng/dl or greater (p = 0.0086) and a similar trend was seen in patients older than 66 years (p = 0.08). There were 62 evaluable patients with a median of 14.9 weeks to DHT 150 pg/ml or greater. There was no association with Gleason score at diagnosis, on study prostate specific antigen, type of prior definitive therapy, or any prior hormonal therapy and time to increase in circulating androgens. CONCLUSIONS: After 6 months of GnRH-A therapy in these patients, DHT and T levels did not return to normal for a median of 14.9 and 16.6 weeks, respectively. PMID- 15821488 TI - Surrogate end point for prostate cancer specific mortality in patients with nonmetastatic hormone refractory prostate cancer. AB - PURPOSE: We determined whether prostate specific antigen (PSA) velocity can serve as surrogate end point for prostate cancer specific mortality (PCSM) in patients with nonmetastatic, hormone refractory prostate cancer. MATERIALS AND METHODS: The study cohort comprised 919 men treated from 1988 to 2002 at 1 of 44 institutions with surgery (560) or radiation therapy (359) for clinical stages T1c-4NxMo prostate cancer, followed by salvage hormonal therapy for PSA failure. All patients experienced PSA defined recurrence while on hormonal therapy. Prentice criteria require that the surrogate should be a prognostic factor and the treatment used did not alter time to PCSM following achievement of the surrogate end point. These criteria were tested using Cox regression. All statistical tests were 2-sided. RESULTS: PSA velocity greater than 1.5 ng/ml yearly was statistically significantly associated with time to PCSM and all cause mortality following PSA defined recurrence while undergoing hormonal therapy (Cox p <0.0001). While initial treatment was statistically associated with time to PCSM and all cause mortality (Cox p = 0.001 and 0.01), this association became insignificant when PSA velocity and potential confounding variables were included in the Cox model (p = 0.22 and 0.93, respectively). The adjusted HR for PCSM in patients who experienced a greater than 1.5 ng/ml increase in PSA within 1 year while on hormonal therapy was 239 (95% CI 10 to 5,549). CONCLUSIONS: These data provide evidence to support PSA velocity greater than 1.5 ng/ml yearly as a surrogate end point for PCSM in patients with nonmetastatic, hormone refractory prostate cancer. Enrolling these men onto clinical trials evaluating the impact of chemotherapy on time to bone metastases and PCSM is warranted. PMID- 15821489 TI - Prevalence of testicular intraepithelial neoplasia in healthy males. AB - PURPOSE: Testicular intraepithelial neoplasia (TIN), also called intratubular germ cell neoplasia, is thought to be the precursor of testicular germ cell tumors (GCTs). Evidence for this theory stems from clinicopathological studies because to our knowledge experimental models are lacking. While GCT clinical risk groups have been studied extensively for TIN, only 1 small study to date has assessed the presence of TIN in the general population. In the current study we analyzed the prevalence of TIN in a large group of healthy men. MATERIALS AND METHODS: Bilateral testicular specimens were obtained from 1,388 presumably healthy men who died unexpectedly and underwent autopsy at forensic institutes in Germany. Median age in this patient sample was 33 years. Specimens were fixed in Stieve's solution and all specimens were examined immunohistologically with placental alkaline phosphatase staining. RESULTS: TIN was found in 6 cases (0.43%, 95% CI 0.16 to 0.94). Two cases also showed microinvasive seminoma. TIN was bilateral in 1 case. The right testicle was afflicted in 4 cases and the left was affected in 1. Median age of the individuals with TIN was 33 years. CONCLUSIONS: As found in the current study, the prevalence of TIN is consistent with the lifetime risk of GCT in Germany. Data in the current study are in accordance with those in a previous study from Denmark. In all these data represent fundamental support of the role of TIN as the precursor of GCT. PMID- 15821498 TI - Comparison of retroperitoneoscopic nephrectomy versus open approaches to nonfunctioning tuberculous kidneys: a report of 44 cases. AB - PURPOSE: We retrospectively investigated the advantages of retroperitoneoscopic nephrectomy for nonfunctioning tuberculous kidneys by comparing its clinical results, operative methods and skills with those of open nephrectomy. MATERIALS AND METHODS: Clinical data on 22 patients with nonfunctioning tuberculous kidneys who underwent retroperitoneoscopic nephrectomy, including simple and subcapsular nephrectomy, were compared with those on 22 who underwent open nephrectomy for a similar indication during the same period. Results in the 2 groups were analyzed. RESULTS: There was no statistical difference between the retroperitoneoscopy and open surgery groups with regard to patient age, sex or mean operative time +/- SD (93.0 +/- 12.6 vs 92.6 +/- 35.5 minutes). Mean blood loss was significantly less in the retroperitoneoscopy group than in the open surgery group (78.3 +/- 60.6 vs 160 +/- 120.0 ml). Mean hospital stay after operation was notably shorter in the retroperitoneoscopy group compared with the open surgery group (3.3 +/- 0.9 vs 9.1 +/- 0.8 days). The mean analgesic requirement for opioids and diclofenac sodium was also lower in the retroperitoneoscopy group than in the open surgery group (0 vs 2.1 +/- 0.9 and 5.2 +/- 1.1 vs 5.8 +/- 1.3 doses, respectively). CONCLUSIONS: Retroperitoneoscopic nephrectomy for renal tuberculosis has several advantages over open nephrectomy, namely a smaller wound, less blood loss and more rapid recovery. It may provide a safe and reliable method for treating refractory renal tuberculosis clinically. PMID- 15821499 TI - Intravesical resiniferatoxin for the treatment of interstitial cystitis: a randomized, double-blind, placebo controlled trial. AB - PURPOSE: Interstitial cystitis is a painful bladder condition of unknown etiology and poorly understood pathophysiology. Current therapies have met with limited success. Vanilloid receptor agonists such as resiniferatoxin (RTX) desensitize C fibers that transmit pain; it is hypothesized that such drugs will be effective in the treatment of interstitial cystitis and painful bladder syndrome by decreasing the pain that leads to urinary frequency and urgency. MATERIALS AND METHODS: A randomized, double-blind, placebo controlled study was conducted in 163 patients with interstitial cystitis. Participants were randomly assigned to receive a single intravesical dose of 50 ml of either RTX 0.01 microM, 0.05 microM, 0.10 microM, or placebo. Safety and efficacy was evaluated over 12 weeks. The primary efficacy endpoint was the Global Response Assessment, a 7-point scale rating overall change in symptoms of interstitial cystitis after 4 weeks. Secondary efficacy endpoints included reduction in pain, urgency, frequency, nocturia, average void volume, and the O'Leary-Sant Symptom and Problem Indexes. RESULTS: RTX did not improve overall symptoms, pain, urgency, frequency, nocturia, or average void volume during 12 weeks followup. RTX resulted in a dose dependent increase in the incidence of instillation pain, but was otherwise generally well tolerated. CONCLUSIONS: In the largest prospective, randomized clinical trial reported to date with intravesical vanilloid therapy, single administration of RTX at doses of 0.01 microM to 0.10 microM was not effective in patients with interstitial cystitis. PMID- 15821500 TI - Can outcome of internal urethrotomy for short segment bulbar urethral stricture be predicted? AB - PURPOSE: We predicted the outcome of visual internal urethrotomy (VIU) by measuring the percentage of lumen narrowing at the stricture site on retrograde urethrography (RGU). MATERIALS AND METHODS: From January 1991 to June 2002 patients with primary bulbar urethral strictures who underwent VIU were selected for the study. Patients with a history of intervention, complete block of the urethral lumen and stricture greater than 2 cm were excluded from study. Urethral diameter at the area of maximum stenosis and at the normal distal urethra was measured on RGU with Vernier caliper and percentage narrowing was derived. Patients were followed 3 times monthly with symptoms, calibration and whenever required with RGU. Recurrence of symptoms, failure to self-calibrate and the need for secondary procedure were considered treatment failure. RESULTS: Complete followup data were available in 105 patients (44 grade 1 and 61 grade 2). Mean bulbar urethral stricture length was 0.86 cm. Inflammation was the cause of stricture in 83 (79%) and trauma the cause in 22 (21%) patients. In the Cox proportional hazards model only grade of narrowing had a significant impact on outcome. There were 41 cases of treatment failure in the total followup of 46 +/- 9 months. Mean recurrence-free duration +/- SD was 13 +/- 15 and 44.52 +/- 19 months in cases of treatment failure and success, respectively (p <0.0001). Mean percentage narrowing was significantly higher with treatment failure (69.9% +/- 16.1% vs 48.55% +/- 17.3%, p <0.0001). A cutoff of 74% for urethral narrowing was derived to predict the outcome with 78% probability. CONCLUSIONS: Percentage narrowing of the urethral lumen at the stricture site is a useful predictor of VIU outcome. PMID- 15821507 TI - The efficacy of dietary intervention on urinary risk factors for stone formation in recurrent calcium oxalate stone patients. AB - PURPOSE: Nutrition is suggested to be the major environmental risk factor in idiopathic calcium oxalate stone disease. The study was designed to evaluate the effect of dietary intervention on urinary risk factors for recurrence in calcium oxalate stone formers. MATERIALS AND METHODS: A total of 76 men and 31 women with idiopathic calcium oxalate stone disease collected 24-hour urine on their habitual, self-selected diets and after 7 days on a balanced standardized diet according to the recommendations for calcium oxalate stone formers. RESULTS: On the usual diet, a urine volume of less than 2.0 l per 24 hours was present in 57.9%, hypercalciuria in 25.2%, hypomagnesuria in 18.7%, hyperoxaluria in 14.0%, hyperuricosuria in 41.3% and hypocitraturia in 57.0% of patients. The frequency of metabolic abnormalities and the risk of calcium oxalate stone formation decreased significantly on the ingestion of the balanced diet, due to the significant increase in urinary volume, pH and citrate excretion and the significant decrease in urinary calcium and uric acid excretion. No change occurred in urinary oxalate and magnesium excretion. CONCLUSIONS: The evaluation of urinary risk profiles of the patients on their usual dietary habits revealed a high risk for calcium oxalate stone formation. A low fluid intake and an increased intake of protein and alcohol were identified as the most important dietary risk factors. The shift to a nutritionally balanced diet according to the recommendations for calcium oxalate stone formers significantly reduced the stone forming potential. PMID- 15821508 TI - Physicochemical metabolic characteristics for calcium oxalate stone formation in patients with gouty diathesis. AB - PURPOSE: We determined why calcium oxalate stones instead of uric acid stones form in some patients with gouty diathesis. MATERIALS AND METHODS: Gouty diathesis was diagnosed from absence of secondary causes of uric acid stones or low urinary pH, and reduced fractional excretion of urate with discriminant score of the relationship between urinary pH and fractional excretion of urate less than 80. From the stone registry 163 patients with gouty diathesis were identified, including 62 with uric acid stones (GD + UA) and 101 patients with calcium oxalate stones (GD + Ca). Metabolic data and 24-hour urinary chemistry study were compared between the 2 groups. RESULTS: Compared with GD + UA, GD + Ca had significantly greater urinary calcium (196 +/- 96 mg per day vs 162 +/- 82 mg per day, p <0.05) and significantly lower urinary citrate (430 +/- 228 vs 519 +/- 288 mg per day, p <0.05), resulting in higher urinary saturation of calcium oxalate. Both groups had low urinary pH (less than 5.5) and high urinary undissociated uric acid (greater than 100 mg/dl). Urinary calcium post-oral calcium load was significantly higher in GD + Ca than in GD + UA (0.227 vs 0.168 mg/dl glomerular filtrate, p <0.001). CONCLUSIONS: Calcium oxalate stones may form in some patients with gouty diathesis due to increased urinary excretion of calcium and reduced excretion of citrate. Relative hypercalciuria in GD + Ca may be due to intestinal hyperabsorption of calcium. PMID- 15821510 TI - Laparoscopic repair of vesicovaginal fistula. AB - PURPOSE: Vesicovaginal fistula may be a complication of urogynecologic surgery. We describe the technique of laparoscopic repair of vesicovaginal fistula as performed at our 2 institutions. MATERIALS AND METHODS: Since August 1998 laparoscopic repair of vesicovaginal fistula was performed in 15 select patients who had clear indications to undergo surgical treatment through an abdominal approach. Hysterectomy had previously been performed in 14 patients (93%). Conservative treatment was initially attempted for more than 2 months in all cases. Four patients had undergone a previous surgical fistula closure attempt with unsuccessful results. Our technique involved cystoscopy, catheterization of the vesicovaginal fistula, laparoscopic cystotomy, opening and excision of the fistulous tract, dissection of the bladder from the vagina, cystotomy closure and colpotomy with interposition of a flap of healthy tissue. Demographic as well as perioperative and outcome data were recorded. RESULTS: Average patient age was 38 years. None of the cases required open conversion. Mean operative time was 170 minutes (range 140 to 240). Mean hospital stay was 3 days (range 2 to 5). The mean duration of bladder catheterization was 10.4 days (range 9 to 15) At a mean followup of 26.2 months (range 3 to 60) 14 patients (93%) were cured. CONCLUSIONS: We believe that laparoscopic repair of vesicovaginal fistula is a feasible and efficacious minimally invasive approach for the management of this entity. PMID- 15821509 TI - Stone and pelvic urine culture and sensitivity are better than bladder urine as predictors of urosepsis following percutaneous nephrolithotomy: a prospective clinical study. AB - PURPOSE: Urosepsis due to manipulation during percutaneous nephrolithotomy (PCNL) can be catastrophic despite prophylactic antibiotic coverage, and negative midstream urine culture and sensitivity testing (C&S). It has been postulated that bacteria in the stone may be responsible for systemic infection. In this prospective study we determined the correlation between different sites of urine sampling, including stones, and also ascertained which is more predictive of urosepsis. MATERIAL AND METHODS: All patients undergoing PCNL who fulfilled our selection criteria were recruited. The samples collected were 1) midstream urine and bladder urine at cystoscopy, 2) renal pelvic urine collected at percutaneous puncture of the pelvicaliceal system and 3) extracted and later fragmented stones. They were sent immediately for C&S. Patients were monitored for systemic inflammatory response syndrome (SIRS). RESULTS: A total of 54 procedures were suitable for analysis. Midstream urine C&S was positive in 11.1% of cases, stone C&S was positive in 35.2% and pelvic C&S was positive in 20.4% (p = 0.009). Pelvic urine C&S predicted infected stones better than bladder urine C&S. Of the patients 37% had SIRS and 3 experienced septic shock. Patients with infected stones or pelvic urine were found to be at a relative risk for urosepsis that was at least 4 times greater (p = 0.0009). Bladder urine did not predict SIRS. Stone C&S had the highest positive predictive value of 0.7. Preoperative hydronephrosis correlated with infected pelvic urine. No patients with urosepsis had positive blood C&S. CONCLUSIONS: The results of this study suggest that positive stone C&S and pelvic urine C&S are better predictors of potential urosepsis than bladder urine. Therefore, routine collection of these specimens is recommended. PMID- 15821517 TI - Venous thrombosis prophylaxis for urological laparoscopy: fractionated heparin versus sequential compression devices. AB - PURPOSE: Venous thromboembolism (VTE) is a significant postoperative complication. Common methods of VTE prophylaxis include subcutaneous fractionated heparin (FH) and lower extremity sequential compression devices (SCD). There is no conclusive evidence supporting 1 method compared to the other for urological laparoscopy. We examined the rates of postoperative hemorrhagic and thrombotic complications after laparoscopic urological procedures in patients treated with FH or SCD as VTE prophylaxis. MATERIALS AND METHODS: A prospective database augmented by retrospective chart review included all patients who underwent urological laparoscopic surgery of the upper retroperitoneum at our institution from June 2000 to December 2002. Patients nonrandomly received FH or SCD as VTE prophylaxis beginning on the day of surgery. RESULTS: A total of 344 patients were included in this study, 172 in the FH group and 172 in the SCD group. Thrombotic complications included VTE and/or pulmonary embolism. Hemorrhagic complications were minor or major, the latter requiring transfusion or other intervention. In both groups the rate of thrombotic complication was 2 of 172 (1.2%). The rate of hemorrhagic complication was 16 of 172 (9.3%) in the FH group, of which 12 of 172 (7.0%) were major. The hemorrhagic complication rate was 6 of 172 (3.5%), with 5 of 172 (2.9%) being major in the SCD group. CONCLUSIONS: After urological laparoscopy of the upper retroperitoneum, subcutaneous fractionated heparin is associated with increased hemorrhagic complications (p = 0.045), without a reduction in thrombotic complications (p >0.999), compared with sequential compression devices. PMID- 15821518 TI - Diagnosis, management and prognosis of vaginal erosion after transobturator suburethral tape procedure using a nonwoven thermally bonded polypropylene mesh. AB - PURPOSE: We studied the diagnosis, management and prognosis of vaginal mesh erosion using a thermally bonded nonwoven polypropylene mesh in a transobturator suburethral tape procedure for the surgical treatment of stress urinary incontinence in women. MATERIALS AND METHODS: A total of 65 patients diagnosed with stress urinary incontinence underwent a transobturator suburethral tape procedure with a fusion welded, nonwoven, nonknitted polypropylene mesh, with or without a central silicone coated section, at our institution. All women were followed and if vaginal erosion was diagnosed, cystoscopy and vaginoscopy were performed, the mesh was partially or completely removed and, if necessary, posterior cough test and urodynamic study were performed. RESULTS: Of the 65 patients 9 (13.8%) were diagnosed with vaginal erosion at the vaginal incision during a relatively long postoperative period (mean 290 days). All presented with vaginal discharge and 1 had a severe complication (obturator abscess). Complete mesh removal was necessary in 8 patients and only 2 (22%) had recurrent stress urinary incontinence. CONCLUSIONS: A 13.8% rate of vaginal mesh erosion using a nonwoven thermally bonded polypropylene mesh was reported. This complication was probably due to the characteristics of the mesh and not to the transobturator approach. Complete removal of the tape is recommended and the continence status prognosis is good (78%). PMID- 15821519 TI - Long-term followup of augmentation enterocystoplasty and continent diversion in patients with benign disease. AB - PURPOSE: We evaluated long-term outcomes in patients undergoing augmentation enterocystoplasty (AC) (with or without an abdominal stoma) or continent urinary diversion in patients with benign urological disorders. MATERIALS AND METHODS: This was a retrospective study of 76 consecutive adults who underwent AC (with or without an abdominal stoma) or continent urinary diversion because of benign urological conditions. The outcomes assessed were a patient satisfaction questionnaire, continence status, catheterization status, bladder capacity, bladder compliance, detrusor instability, maximum detrusor pressure, upper tract status, significant postoperative morbidity, need for reoperation, persistent diarrhea and vitamin B12 deficiency. RESULTS: The 76 patients (18 men and 58 women) were 19 to 80 years old (mean age 49). Followup was 1 to 19 years (mean 8.9). Preoperative diagnoses were neurogenic bladder in 41 patients, refractory detrusor overactivity in 9, interstitial cystitis in 7, end stage bladder disease in 7, radiation cystitis in 3, exstrophy in 3, postoperative urethral obstruction in 3 and low bladder compliance in 3. A total of 50 patients underwent simple AC, 15 underwent AC with an abdominal stoma and 11 underwent continent supravesical diversion. Of the 71 evaluable patients 49 (69%) considered themselves cured, 14 (20%) considered themselves improved and 8 (11%) considered treatment to have failed. All 7 patients with interstitial cystitis had failed treatment. Mean bladder capacity increased from 166 to 572 ml and mean maximum detrusor pressure decreased from 53 to 14 cm H2O. Serum creatinine improved or remained normal in all patients. Five patients experienced persistent diarrhea requiring intermittent antispasmodics but none had vitamin B12 deficiency, pernicious anemia or malabsorption syndrome. Long-term complications were stomal stenosis or incontinence in 11 of 26 patients (42%) with stomas, de novo bladder and renal stones in 2 of 71 (3%) and 1 of 71 (1%), respectively, and recurrent bladder stones in 6%. Small bowel obstruction occurred in 5 of 71 patients (7%), requiring surgical exploration in 4 (6%). CONCLUSIONS: AC and urinary diversion provide a safe and effective long-term therapy in patients with refractory neurogenic bladder but stomal problems in patients with continent diversion continue to be a source of complications. PMID- 15821526 TI - The overactive bladder-symptom composite score: a composite symptom score of toilet voids, urgency severity and urge urinary incontinence in patients with overactive bladder. AB - PURPOSE: To our knowledge there is no index in urology that yields a single, quantifiable and clinically interpretable measure of overactive bladder (OAB) symptoms, including urgency, 24-hour voiding frequency and urge urinary incontinence (UUI). Urgency is the most difficult of these symptoms to measure. The Indevus Urgency Severity Scale (Indevus Pharmaceuticals, Lexington, Massachusetts) was recently developed and validated to capture urgency severity per toilet void. The scale has been combined with 24-hour frequency and UUI episodes to create the OAB Symptom Composite Score (OAB-SCS). We present this composite score. MATERIALS AND METHODS: Two multicenter trials were performed to determine the effects of trospium chloride given as 20 mg tablets vs placebo. A total of 1,157 patients, including 581 who received placebo and 576 who received trospium, were randomly assigned to treatment. Daily OAB-SCS totals were obtained for each patient for each day during the 7-day diary collection period for every visit. RESULTS: The average baseline OAB-SCS value was 36. The mean change from baseline in the trospium and placebo groups was 5 and 1 OAB-SCS points in patients with mild OAB, 10 and 5 in patients with moderate OAB, and 13 and 9 in patients with severe OAB, respectively. CONCLUSIONS: The OAB-SCS discriminated between placebo and pharmacologically treated (trospium chloride) patients with OAB in this study. The OAB-SCS is an improvement over individual symptoms alone. It is easy to implement and interpret and it will prove to be a clinically relevant tool in clinical trials in which patient diary data are captured. PMID- 15821527 TI - Functional magnetic stimulation for mixed urinary incontinence. AB - PURPOSE: In this study we determined the efficacy of functional magnetic stimulation (FMS) compared to placebo for treating women with mixed urinary incontinence (MUI). MATERIALS AND METHODS: A total of 39 women with MUI were randomly assigned to the FMS group (23 patients) or to the placebo group (16 patients). FMS was applied continuously at 18.5 Hz day and night for 2 months. Conventional urodynamic studies were performed before and after stimulation. Outcome measures assessed were clinical (daytime frequency, nocturia, pad use, pad weight) and urodynamic variables (first sensation of bladder filling, maximum cystometric capacity, maximum urethral closure pressure), and patient subjective assessment (visual analogue scale). RESULTS: After 2 months of FMS significant decreases in voiding frequency (from 9.0 to 6.7, p = 0.0002), nocturia (from 2.6 to 1.4, p = 0.0007) and pad use (from 3.9 to 2.2, p = 0.007) were observed only in the FMS group. First sensation of bladder filling and maximum cystometric capacity increased significantly after stimulation compared with prestimulation levels only in the FMS group, p = 0.003 (from 118 to 174 ml) and p = 0.00004 (from 267 to 396 ml), respectively. A total of 18 women (78.3%) reported an improvement in symptoms after FMS with an average success rate of 41.9%. The success rate was significantly lower in the placebo group (p = 0.021) at 22.9%. CONCLUSIONS: Functional magnetic stimulation was useful and safe for treating women with MUI. PMID- 15821528 TI - A randomized controlled trial of duloxetine alone, pelvic floor muscle training alone, combined treatment and no active treatment in women with stress urinary incontinence. AB - PURPOSE: We primarily compared the effectiveness of combined pelvic floor muscle training (PFMT) and duloxetine with imitation PFMT and placebo for 12 weeks in women with stress urinary incontinence (SUI). In addition, we compared the effectiveness of combined treatment with single treatments, single treatments with each other and single treatments with no treatment. MATERIALS AND METHODS: This blinded, doubly controlled, randomized trial enrolled 201 women 18 to 75 years old with SUI at 17 incontinence centers in the Netherlands, United Kingdom and United States. Women averaged 2 or more incontinence episodes daily and were randomized to 1 of 4 combinations of 80 mg duloxetine daily, placebo, PFMT and imitation PFMT, including combined treatment (in 52), no active treatment (in 47), PFMT only (in 50) and duloxetine only (in 52). The primary efficacy measure was incontinence episode frequency. Other efficacy variables included the number of continence pads used and the Incontinence Quality of Life questionnaire score. RESULTS: The intent to treat population incontinence episode frequency analysis demonstrated the superiority of duloxetine with or without PFMT compared with no treatment or with PFMT alone. However, pad and Incontinence Quality of Life analyses suggested greater improvement with combined treatment than single treatment. A completer population analysis demonstrated the efficacy of duloxetine with or without PFMT and suggested combined treatment was more effective than either treatment alone. CONCLUSIONS: The data support significant efficacy of combined PFMT and duloxetine in the treatment of women with SUI. We hypothesize that complementary modes of action of duloxetine and PFMT may result in an additive effect of combined treatment. PMID- 15821529 TI - Long-term results of the bulbourethral sling procedure. AB - PURPOSE: We evaluated the long-term efficacy of the male bulbourethral sling procedure in the treatment of post-radical prostatectomy urinary incontinence. MATERIALS AND METHODS: Between October 1994 and June 2000, 95 patients with post radical prostatectomy incontinence underwent bulbourethral sling placement with tetrafluoroethylene bolsters at our hospital. Ultimately 71 of these patients responded to our questionnaire and they were classified into 2 groups. Group 1 consisted of 62 patients who had not undergone prior radiation therapy and group 2 consisted of 9 who had undergone radiation therapy before the sling procedure. Patients were asked to respond to questions regarding continence status as well as the validated Incontinence Quality of Life and International Prostate Symptom Score questionnaires. RESULTS: Mean followup from the most recent sling procedure was 4 years (range 0.27 to 6.55). Average patient age at questionnaire response was 74 years. A total of 86 procedures were performed on 71 patients. Of the 71 patients 7 underwent either sling removal or artificial urinary sphincter placement and were excluded from questionnaire analysis. Including retightening procedures 68% of the patients (72% of group 1, 43% of group 2) required 2 or less pads daily. Of the patients 36% (42% of group 1 and 14% of group 2) required 0 pads. CONCLUSIONS: The male bulbourethral sling procedure remains an effective treatment for post-prostatectomy incontinence at 4-year followup. PMID- 15821530 TI - The male sling for post-prostatectomy incontinence: mean followup of 18 months. AB - PURPOSE: We review our experience with 42 consecutive patients undergoing the bone anchored male sling procedure. MATERIALS AND METHODS: A retrospective chart review was performed, and complete data and followup were available in 38 of the 42 patients. Success was defined as wearing 1 thin pad or less per day, or social continence. Variables such as severity of incontinence, age, detrusor overactivity, previous artificial urinary sphincter, history of radiation and intraoperative flow pressures were analyzed for success and risk of failure. RESULTS: With a mean followup of 18 months (range 6 to 26), 39.5% (15 of 38) were considered a success (socially continent). A statistically significant trend in the degree of preoperative incontinence predicting success was identified. Social continence in mild, moderate and severe cases was achieved in 67%, 50% and 0%, respectively (p =0.001/95% CI). Only 15.8% of patients were completely dry and not wearing pads. Significant perineal pain was reported in the early postoperative period but resolved in all patients. Infection occurred in 3 patients with erosion found in 1. CONCLUSIONS: Although these results are not as encouraging as previous reports, carefully selected patients with mild to moderate incontinence are good candidates for the male sling. Patients with a history of radiation, previous artificial urinary sphincter or severe incontinence should be counseled about the higher risk of treatment failure. Patients should be informed of the possibility of progressive failure with time and the occurrence of significant perineal pain in the early postoperative period. PMID- 15821545 TI - Sexual function in women with pelvic organ prolapse compared to women without pelvic organ prolapse. AB - PURPOSE: We compared sexual function in women with pelvic organ prolapse to that in women without prolapse. MATERIALS AND METHODS: We collected sexual function data using a standardized, validated, condition specific questionnaire. The study group consisted of 30 women with pelvic organ prolapse and it was compared with 30 unmatched controls without evidence of prolapse. RESULTS: The 2 groups were similar in age, race, parity and postmenopausal hormone use. Subjects in the study group were more likely to have undergone previous pelvic surgery. Mean total Pelvic Organ Prolapse/Urinary Incontinence Sexual Function Questionnaire scores +/- SD were lower in the study group compared with controls (81.4 +/- 7.3 vs 106.4 +/- 15.5, p <0.001). In the study group total questionnaire scores in women with prior pelvic surgery were similar to those in women without prior pelvic surgery (79.3 +/- 14.9 vs 82.9 +/- 10.2, p = 0.61). CONCLUSIONS: Pelvic organ prolapse appears to have a significant negative impact on sexual function. PMID- 15821546 TI - Histological alterations in cavernous tissue after radical prostatectomy. AB - PURPOSE: Radical prostatectomy often results in erectile dysfunction because of lesions to the erectile nerves. In this study we evaluated histomorphological alterations in cavernous smooth muscle and collagen content after radical prostatectomy. MATERIALS AND METHODS: A total of 19 patients between 57 and 69 years old with prostate adenocarcinoma and normal erectile function, as reported and validated by RigiScan (UroHealth Systems, Laguna Niguel, California) testing, underwent corpora cavernosa biopsy in the operating room before radical prostatectomy, and 2 and 12 months after surgery. No patient underwent hormone therapy before or after surgery and none was diabetic. Elastic fibers (manual counting), muscle specific actin (immunostaining) and collagen content (computerized morphometric imaging) were measured in the 3 biopsies. RESULTS: In all cases the first postoperative histological assessment revealed some disorganization. Trabecular elastic fibers (p <0.0003) and smooth muscle fibers were decreased and collagen content was significantly increased (p <0.0003) compared with preoperative biopsies. One year after surgery elastic fibers (p <0.0003) and smooth muscle fibers were decreased and collagen content was significantly increased (p <0.0003) compared with the first postoperative biopsy. Moreover, organized collagen and trabecular protocollagen deposits were increased. CONCLUSIONS: Progressive fibrosis in the corpora cavernosa after radical prostatectomy probably results from denervation and/or an ischemic process, which is caused in turn by the ligation of anomalous pudendal artery branches or of venous plexuses that drain to or from the corpora cavernosa. Fibrosis and the subsequent loss in elasticity and function of erectile tissue probably together cause erectile dysfunction. PMID- 15821547 TI - No-needle jet anesthetic technique for no-scalpel vasectomy. AB - PURPOSE: We describe a new, modified jet injection technique for local anesthesia for no-scalpel vasectomy without the use of a needle, which may minimize the fear of vasectomy in men due to the needle involved in local anesthesia. MATERIALS AND METHODS: A MadaJet Medical Injector (MADA Medical Products, Carlstadt. New Jersey) was used in this study to deliver a high pressure spray of 0.1 cc local anesthetic solution directly through the scrotal skin down onto the tissue around the vas. Two or 3 jet injections are delivered to each vas and a total of 4 to 6 suffice for the entire vasectomy. RESULTS: No-needle jet injection is remarkably effective for local anesthesia for no-scalpel vasectomy. The average volume of anesthetic solution per jet injection is 0.1 cc with 0.2 to 0.3 cc for each vas. Onset is almost immediate, within 10 to 20 seconds after injection. About 465 patients were anesthetized by the jet injection technique with great satisfaction. The average visual analog scale score for the pain of the jet injection itself was 1.71 of 10. The average visual analog scale score for the pain of subsequent vasectomy during the surgical procedure was 0.66 of 10 (median 0.2). No hematomas were noted. Patients experience the mild discomfort of a pinch, not unlike a rubber band, with the first injection. CONCLUSIONS: No-needle anesthesia with jet injection is a new technique to deliver rapid onset of profound local anesthesia to the patient undergoing vasectomy. It is a simple and safe approach with high patient satisfaction, as reflected in low pain scores. The benefit of this technique without a needle is that it may decrease the fear of pain in men and enhance the popularity of vasectomy worldwide. PMID- 15821548 TI - Model to predict if a vasoepididymostomy will be required for vasectomy reversal. AB - PURPOSE: We devised a model to predict, preoperatively, the need for a vasoepididymostomy (VE) when performing a vasectomy reversal. Urologists could use it to identify those patients who need a referral to an experienced VE surgeon. MATERIALS AND METHODS: We performed a retrospective review of 483 patients who underwent vasectomy reversal by a single surgeon (AJT) including 393 vasovasostomies and 90 vasoepididymostomies. Selection was based on chart availability. Established criteria were used in deciding the type of reversal (eg gross appearance and microscopic examination of vasal fluid). Type of reversal, patient age and time since vasectomy were recorded. Univariate analysis revealed that patient age (p <0.001) and time since vasectomy (p <0.001) were significant predictors of reversal type. On multivariate logistic regression analysis, time since vasectomy (p <0.001) was the only significant independent predictor. We designed a linear regression algorithm based on time since vasectomy and patient age to predict if a VE would be performed. The model was designed using 433 patients and then tested on a separate randomly selected 50 patient group. The model was designed to be 100% sensitive in detecting patients requiring VE. RESULTS: In the test group the model was 100% sensitive in predicting VE with a specificity of 58.8%. The area under the ROC curves for the design and test groups was 0.8. Palm (PalmSource Inc., Sunnyvale, California) and Windows (Microsoft Corporation, Redmond, Washington) versions are available as free shareware from www.uroengineering.com. CONCLUSIONS: The model is 100% sensitivity in detecting those patients who may require a VE during vasectomy reversal (specificity of 58.8%). It may allow urologists to preoperatively identify these patients. PMID- 15821559 TI - Exploring the learning curve, pathological outcomes and perioperative morbidity of laparoscopic partial nephrectomy performed for renal mass. AB - PURPOSE: We investigated the learning curve, pathological results and perioperative morbidity of laparoscopic partial nephrectomy (LPN). MATERIALS AND METHODS: The records of all LPN cases at our institution between January 1999 and March 2004 were reviewed. Of 223 cases 217 (97.3%) were performed for an enhancing renal mass. RESULTS: Mean tumor size was 2.6 cm (range 1 to 10) and 95.4% of patients had a normal contralateral kidney. Transient vascular control was performed in 75.1% of cases. Mean operative time (186 minutes) decreased with surgeon experience (p = 0.003) but was independent of tumor size (p = 0.964). Mean warm ischemia time (27.6 minutes) depended on tumor size (p = 0.005) but not on experience (0.964). Mean blood loss was 385 cc and the perioperative transfusion rate was 6.9%. Postoperative complications occurred in 23 cases (10.6%) with the most common being ileus (1.8%), bleeding (1.8%) and urinary leakage (1.4%). Although the mean serum creatinine change after LPN was a function of tumor size (p <0.001), it was clinically insignificant (0.13 mg/dl). No significant relationship was observed between warm ischemia time and creatinine change (p = 0.262). The final pathological evaluation revealed renal cell carcinoma in 144 patients (66.4%) and the overall positive margin rate was 3.5%. Only 2 renal cell carcinoma recurrences in the operated kidney (1.4%) were identified (mean followup +/- SD 24 +/- 12 months). CONCLUSIONS: LPN is an effective approach for treating small renal masses with low perioperative morbidity. Contrary to previous reports, more than 30% of the enhancing renal lesions excised in this series were found to be benign on final pathological evaluation. PMID- 15821560 TI - Impact of hospital and surgeon volume on in-hospital mortality from radical cystectomy: data from the health care utilization project. AB - PURPOSE: We determined the influence of hospital and surgeon volume on various outcome parameters after radical cystectomy for bladder cancer. MATERIALS AND METHODS: All inpatient discharges after radical cystectomy for bladder cancer (1988 to 1999) from the Health Care Utilization Project-Nationwide Inpatient Sample were included in the analysis. Hospital and individual surgeon volume of discharges per year were separated into terciles. Outcome measures were in hospital mortality, length of stay (LOS), and inflation adjusted charge per admission. Mortality was compared among hospital volume levels using the Mantel Haenszel chi-square test while the LOS and charges were compared using ANOVA. Multivariate linear and logistic regression analyses were used to adjust for confounding factors. All the analyses were also performed in 3 different age strata (younger than 50 years, 50 to 69 years and 70 years or more). RESULTS: There were 13,964 patients who underwent radical cystectomy. Overall in-hospital mortality was 408 of 13,964 (2.9%), average LOS was 14 days (+/- SD 10.28) and average charges were 47,146 dollars (+/- SD 45,263 dollars). In-hospital mortality was significantly associated with higher volume particularly for patients older than 50 years. Surgeon volume did not influence in-hospital mortality except for patients in the 50 to 69-year-old age group. Results of multivariate regression analysis demonstrated hospital volume was a significant predictor of in-hospital mortality but this effect was lost when controlling for surgeon volume. LOS was significantly higher for low volume surgeons. High volume hospitals had lower average total charges compared with the low and moderate volume hospitals. CONCLUSIONS: Hospital and surgeon volume have a significant impact on in-hospital mortality and LOS after radical cystectomy. Radical cystectomy performed at a higher volume center may result in lower charges and shorter hospital stay while decreasing the likelihood of in-hospital mortality. PMID- 15821561 TI - 5-year urinary and sexual outcomes after radical prostatectomy: results from the prostate cancer outcomes study. AB - PURPOSE: Prior studies of postoperative outcomes following radical prostatectomy have been limited by selection bias and short-term followup. In this study we assessed temporal changes in urinary and sexual function up to 5 years following radical prostatectomy in a population based cohort. MATERIALS AND METHODS: A sample of 1,288 men with localized prostate cancer who underwent radical prostatectomy and completed a baseline survey within 6 to 12 months of diagnosis were included in the analysis. Two and 5-year functional and quality of life data were collected, as was information on the use of erectile aids. Temporal functional changes and potentially confounding or modifying factors were assessed using longitudinal regression models. RESULTS: Of these men 14% reported frequent urinary leakage or no urinary control 60 months after diagnosis, which was slightly higher than the 10% reporting incontinence at 24 months (p = 0.007). At 60 months 28% of the men had erections firm enough for intercourse compared with 22% at 24 months (p = 0.003). Sildenafil was the most commonly used erectile aid (43% ever used) and 45% of users reported that it helped "somewhat" or "a lot." CONCLUSIONS: Urinary and sexual dysfunction were common 5 years following radical prostatectomy in this large, community based cohort of prostate cancer survivors. While a small minority of subjects experienced changes in urinary or sexual function between years 2 and 5 after prostatectomy, functional outcomes remained relatively stable in the majority of participants. PMID- 15821565 TI - Retroperitoneal laparoscopic versus open pyeloplasty in children. AB - PURPOSE: The indications for laparoscopy in pediatric urology are expanding and yet the advantages over open surgery remain unclear. We compared the results of retroperitoneal laparoscopic vs open pyeloplasty for pyeloureteral junction obstruction in children. MATERIALS AND METHODS: A total of 22 children with a mean age of 88 months (range 25 to 192) underwent laparoscopic dismembered pyeloplasty via the retroperitoneal approach. An additional 17 children with a mean age of 103 months (range 37 to 206) underwent similar procedures via open surgery through a flank incision. We retrospectively analyzed and compared operative time, the use of analgesics (acetaminophen or morphine derivatives) and hospital stay. RESULTS: The 2 groups were similar in mean age and weight at surgery. Mean operative time was significantly shorter in the open surgery vs the laparoscopy group (96 minutes, range 50 to 150 vs 219, range 140 to 310, p <0.0001). Mean postoperative use of acetaminophen (1.9 vs 3.22 days, p = 0.03) and morphine derivatives (1.9 vs 3.06 days, p not significant) was less in the laparoscopy group. Mean hospital stay was shorter in the laparoscopy group than in the open surgery group (2.4 days, range 1 to 5 vs 5, range 3 to 7, p <0.0001). Mean followup was 21 (range 12 to 51) and 24 months (range 12 to 60) in the open and laparoscopy groups, respectively. CONCLUSIONS: The operative time of laparoscopic pyeloplasty remains significantly longer than that of the open procedure in children. The main advantage of the laparoscopic approach is that it significantly decreases hospital stay compared with that after an open procedure. Although in our study analgesic use was less after laparoscopy, our results should be confirmed by a prospective, randomized study. PMID- 15821566 TI - Buried penis: congenital syndrome of a short penile shaft and a paucity of penile shaft skin. AB - PURPOSE: The term "buried penis" has been applied to a variety of penile abnormalities and includes an apparent buried penis that is obvious at birth. The purpose of this study was to examine prospectively the congenital buried penis and to evaluate an operative technique for its management. MATERIALS AND METHODS: A total of 31 males 2 to 28 months old (mean age 12.3 months) with a congenital buried penis underwent surgical correction of the anomaly. Measurements were made of the penile shaft skin, inner leaf of the prepuce, glans length and stretched penile length. Observations of the subcutaneous tissue of the penis were made. The outer leaf of the prepuce was resected, following which covering of the penile shaft was accomplished with a combination of the penile shaft skin and the inner leaf of the prepuce. RESULTS: Stretched penile lengths ranged from 2.3 to 4.1 cm (mean 3.1). The glans length from the tip of the glans dorsally to the corona ranged from 0.9 to 1.6 cm (mean 1.2). The inner leaf of the prepuce ranged from 0.9 to 2.2 cm (mean 1.5) in length, while the dorsal penile skin lengths were 1 to 1.6 cm (mean 0.8). In all patients complete shaft coverage was accomplished using a combination of varying degrees of penile shaft skin and inner leaf of the prepuce. In no case was there a requirement for either unfurling of the inner and outer leaf of the prepuce or mobilization of scrotal flaps to accomplish shaft coverage. All patients healed well and have done well with a followup of 6 months to 1 year. CONCLUSIONS: Congenital buried penis is a syndrome consisting of a paucity of penile shaft skin and a short penile shaft. The anomaly may be successfully repaired by carefully preserving a length of inner leaf of the prepuce sufficient to cover, in some instances, the length of the penile shaft. Anchoring of the penile skin to the shaft is not recommended. PMID- 15821567 TI - Split onlay skin flap for the salvage hypospadias repair. AB - PURPOSE: Following failed hypospadias repair absence of the penile foreskin, a shortage of ventral skin and residual chordee may all contribute to poor long term results. We describe a technique called the split onlay skin (SOS) flap that has improved our surgical outcomes in boys requiring salvage hypospadias repair. MATERIALS AND METHODS: The SOS flap uses a transverse island of penile shaft skin that is mobilized on its vascular pedicle and rotated into position to the ventrum of the penis at the site of the urethral defect. The flap is transected transversely, and half of the flap is used as an onlay to repair the urethra and the other half is used for additional skin coverage where needed on the penile shaft. We treated 11 boys 30 to 124 months old (mean age 60.3 months) who had a mean of 2 previous failed hypospadias repairs. All 11 boys presented with complex combinations of urethrocutaneous fistulas, stricture or urethral diverticula. RESULTS: Of the 11 patients 6 (54.5%) had development of postoperative fistulas. Five of these fistulas were surgically closed with no further complications. One penoscrotal fistula closed spontaneously after 7 months. Mild chordee from contraction of the flap and a urethral diverticulum developed in 1 boy. At a mean followup of 24.5 months all patients, including those who underwent closure of the secondary fistula, were voiding well with excellent appearance. CONCLUSIONS: In cases where little local tissue is present the SOS flap procedure is an excellent way to transfer healthy dorsal tissue to the ventrum for an onlay salvage urethroplasty while providing additional coverage of the urethral defect and a tension-free skin closure. Despite the high fistula rate we encountered following the initial SOS procedure, we endorse this technique because the transferred dartos provides additional tissue, which facilitates subsequent fistula repair. These boys can achieve a successful cosmetic result without incorporation of scrotal tissue or a free graft, which we believe leads to more predictable results. PMID- 15821568 TI - Long-term followup of bilateral high (sober) urinary diversion in patients with posterior urethral valves and its effect on bladder function. AB - PURPOSE: Although valve ablation is the treatment of choice in patients with posterior urethral valves (PUV), temporary high (ureterostomy) diversion remains controversial. In this study we evaluated the effect of bilateral Sober high urinary diversion on renal and bladder function. MATERIALS AND METHODS: We retrospectively reviewed the records of 36 patients with PUV who underwent bilateral Sober-type diversion. Following valve ablation urodynamic studies were done in all patients after diversion and repeated after ureterostomy closure. The mean duration of diversion was 55 months. Vesicoureteral reflux (VUR), renal dysplasia, serum creatinine during followup and urodynamic parameters were recorded. RESULTS: All patients underwent endoscopic valve ablation. Renal function impairment at the end of followup was present in 15 patients. Renal dysplasia was found in 10 patients, while VUR was present in 16 at the time of the first urodynamic studies. Urodynamic studies after ureterostomy closure showed well preserved bladder capacity or compliance in 80% and 69% of cases, respectively. About a third of patients showed increased capacity and normal compliance. Univariate analysis showed that VUR, urinary tract infection and renal dysplasia significantly correlated with impaired renal function. On multivariate analysis renal dysplasia was an independent prognostic factor for poor prognosis. CONCLUSIONS: In patients with PUV temporary high diversion of the Sober type does not have a negative influence on bladder function. It immediately releases high intrarenal pressures but only improves renal function temporarily and may contribute to postpone the time of end stage renal failure. Renal dysplasia dictates long-term renal outcomes in this group. PMID- 15821569 TI - Vesicoureteral reflux and complete ureteral duplication. Conservative or surgical management? AB - PURPOSE: The management of vesicoureteral reflux (VUR) in a duplicated collecting system (DS) is controversial. Available literature is sparse and it does not assess all co-factors. We compared the outcome of VUR in DS cases with VUR in single system (SS) cases, given similar clinical management in the 2 groups. MATERIALS AND METHODS: A retrospective cohort study was done to compare the outcome of VUR in 35 children (44 units) with a DS and in 95 (150 units) with an SS. Patients with secondary reflux, incomplete duplication and ureterocele were excluded. Two groups were similar in terms of prognostic factors. Mean followup in the DS and SS groups was 43 and 48 months, respectively. Outcomes of interest were spontaneous resolution or uncomplicated persistence vs surgical correction. The same criteria were used to intervene surgically in the 2 groups. RESULTS: In the DS group 24 of 35 patients (69%) underwent surgery compared with 25 of 95 (25%) in the SS group. Multivariate analysis showed an association between surgery due to complications and the presence of DS (p = 0.0001). Higher grade and female gender were independent predictors (p = 0.02 and 0.0007, respectively). The likelihood of undergoing surgery was the same for low grade VUR (I and II) in the DS and SS groups (p = 0.16). No resolution was seen in the duplex group with severe VUR at a mean followup of 42 months. CONCLUSIONS: Low grade VUR in DS cases has an outcome similar to that of VUR in SS cases. Females with moderate or high grade VUR and a DS should be treated differently because they present more often with complications. PMID- 15821570 TI - Contemporary epidemiology of bladder exstrophy in the United States. AB - PURPOSE: Although bladder exstrophy is much discussed in the urology literature, there are few population based epidemiological data available for this rare condition. The purpose of this study was to use a large nationwide database to collect contemporary data on the incidence and demographics of bladder exstrophy. MATERIALS AND METHODS: The Healthcare Cost and Utilization Project Nationwide Inpatient Sample is a 20% sample of nonfederal United States hospitals containing data on 5 million to 7 million inpatient stays per year. The sample was limited to newborns, and International Classification of Disease-9 codes were used to identify cases of bladder exstrophy. We then determined nationally weighted incidence through time, and performed multivariate analyses to identify factors associated with exstrophy. RESULTS: We identified 205 patients with exstrophy among 9,452,110 newborns. The overall weighted incidence of exstrophy was 2.15 per 100,000 live births. The male-to-female ratio was almost even (OR 0.989, 95% CI 0.88 to 1.12). White infants were significantly more likely to present with exstrophy than nonwhites (incidence 2.63 vs 1.54 per 100,000, p <0.0001). Exstrophy incidence also varied by geographic region, socioeconomic status (SES) and insurance status. On multivariate analysis the racial variation in exstrophy incidence persisted even after adjustment for geographic region, SES and insurance status. Conditions such as spina bifida, cleft palate, preterm birth and gastrointestinal anomalies were more common in newborns with exstrophy. CONCLUSIONS: Bladder exstrophy is rare, occurs in equal numbers of live male and female newborns, and is associated with certain co-morbid conditions. Incidence appears to be stable through time. Nonwhite race, uninsured status, high or low SES and Western geographic region are associated with lower exstrophy incidence. PMID- 15821571 TI - Variants of the exstrophy complex: a single institution experience. AB - PURPOSE: Variants of the bladder/cloacal exstrophy complex are rare. Different presentations and subsequent management and outcome are discussed. MATERIALS AND METHODS: We performed a retrospective review of our database of more than 815 patients with the exstrophy complex. Patients with variants of classic epispadias or bladder or cloacal exstrophy were identified. Anatomical presentation, surgical management, type of continence procedures and final outcome were evaluated. RESULTS: Of the 25 patients with variants 13 were treated primarily at our institution and 12 were referred. Time until primary bladder closure ranged from 1 day to 4 years. Followup after continence procedure ranged from 1 month to 39 years. Seven of the 25 patients are awaiting a continence procedure. Six patients are dry without a continence procedure, of whom 4 have superior vesical fistulas. A total of 11 patients underwent bladder neck reconstruction (BNR), of whom 3 are dry, 2 are dry during the day but are wet at night, 1 had a failed procedure and 5 are dry after continent diversion (CD). One additional patient underwent CD initially and is dry. Referred cases of epispadias with bladder prolapse were not recognized at birth and had delayed closure. Impaired bladder growth or failed BNR required CD in 4 patients, and 2 are awaiting a continence procedure. Skin covered and duplicate exstrophy had comparable outcomes to the classic presentations. Duplicated organs were used for reconstructive procedures. Of the 6 patients with cloacal variant 2 are continent of stool and 2 await a Pena procedure. One of these patients has an ileal stoma and 1 has a colostomy. CONCLUSIONS: The initial presentation of exstrophy variants can be confusing, often delaying initial treatment. Superior vesical fistulas permit continence without BNR due to an intact urinary sphincter. Variants such as epispadias with bladder prolapse and duplicate or skin covered exstrophy should be closed at birth with standardized techniques to promote bladder growth for later BNR. These cases are faced with the same long-term problems as the classic presentation. Cloacal variants can present with intact anal innervation, allowing a later Pena procedure. PMID- 15821572 TI - Fetal development of the female external urinary sphincter complex: an anatomical and histological study. AB - PURPOSE: We investigated the fetal development of the smooth (lissosphincter) and striated (rhabdosphincter) female external urinary sphincter. Growth and organization of the muscle fibers around the urethra and morphological modifications due to the development of the vagina were analyzed in detail. MATERIALS AND METHODS: A total of 28 human female fetal specimens were investigated in an anatomical and histological study. The sections were processed according to plastination technology. This technique allows examination of structures and organs of the small pelvis with minimal artifacts in all 3 planes. RESULTS: At gestational week 9 the primordium of the external urethral sphincter complex was observed extending along the anterior aspect of the urogenital sinus, before the development of the primitive urethra and the vaginal primordium. From 15 weeks of gestation the lissosphincter and rhabdosphincter could be identified and clearly distinguished. After 20 weeks of gestation both elements acquired an omega-shaped configuration with a narrow posterior connective tissue raphe that was constantly present, fixing both components to the ventral vaginal wall. Both muscles were mainly located in the middle third of the urethra. In the proximal third of the urethra growth of the vagina led to disappearance of the striated muscle fibers of the rhabdosphincter, whereas the lissosphincter seemed to intermingle with the internal layer of the detrusor musculature of the bladder. CONCLUSIONS: The important morphological characteristics of the female adult rhabdosphincter and lissosphincter (omega-shaped configuration, presence of a narrow connective tissue raphe posteriorly and maximum thickness in the middle third of the urethra) are already evident early in fetal development and do not evolve during postnatal growth or by the influence of sex hormones. PMID- 15821573 TI - Renal outcome following heminephrectomy for duplex kidney. AB - PURPOSE: We evaluated functional outcome following heminephrectomy in duplex kidneys. MATERIALS AND METHODS: A total of 101 patients were identified who underwent upper or lower pole heminephrectomy between January 1995 and December 2000. We retrospectively reviewed 60 case notes with complete data. A total of 37 cases (62%) were diagnosed prenatally, while 20 (33%) presented as single or recurrent urinary tract infection and 3 (5%) as urinary incontinence. Of the patients 52 (87%) had unilateral duplex and 8 (13%) had bilateral duplex kidneys. In addition, 20 of the 60 patients (33%) had ureterocele. Mean age at surgery was 46 months (range 3 to 10 years). RESULTS: Mean function as assessed by nuclear renography of the duplex kidney before heminephrectomy was 39.9% (range 22% to 61%) and following heminephrectomy was 33.1% (13% to 60%). Mean decrease in function was 6.8%, with a decrease in function of less than 5% in 18 patients, 5% to 10% in 13 and more than 10% in 5. The functional values were increased by 1% to 5% in 9 patients and unchanged in 15 patients after heminephrectomy. CONCLUSIONS: Following heminephrectomy there was a significant decrease in 8% of patients and a small decrease in function in the remaining moiety in 51%. PMID- 15821574 TI - Magnetic resonance angiography for the nonpalpable testis: a cost and cancer risk analysis. AB - PURPOSE: For the unilateral nonpalpable testis standard management is open surgical or laparoscopic exploration. An ideal imaging technique would reliably identify testicular nubbins and safely allow children to forgo surgical exploration without compromising future health or fertility. Our goal was to perform a cost and risk analysis of magnetic resonance angiography (MRA) for unilateral nonpalpable cryptorchid testes. MATERIALS AND METHODS: A search of the English medical literature revealed 3 studies addressing the usefulness of MRA for the nonpalpable testicle. We performed a meta-analysis and applied the results to a hypothetical set of patients using historical testicular localization data. Analysis was then performed using 3 different management protocols-MRA with removal of testicular nubbin tissue, MRA with observation of testicular nubbin tissue and diagnostic laparoscopy. A cancer risk and cost analysis was then performed. RESULTS: MRA with observation of testicular nubbin tissue results in 29% of patients avoiding surgery without any increased cost of care. Among the 29% of boys with testicular nubbins left in situ and observed the highest estimated risk was 1 in 300 of cancer developing, and 1 in 5,300 of dying of cancer. CONCLUSIONS: A protocol using MRA with observation of inguinal nubbins results in nearly a third of boys avoiding surgical intervention at a similar cost to standard care without any significant increased risk of development of testis cancer. PMID- 15821575 TI - Lymphatic sparing laparoscopic varicocelectomy: a microsurgical repair. AB - PURPOSE: The division of lymphatic vessels during pediatric varicocelectomy is complicated by hydrocele formation, testicular hypertrophy due to intratesticular edema and decline in testicular function. To prevent these complications, we introduced a microsurgical lymphatic sparing dissection into laparoscopic varicocelectomy. MATERIALS AND METHODS: We retrospectively compared outcomes in 104 boys who underwent microsurgical laparoscopic repair for grade II to III varicocele between April 1999 and December 2002 to a group of 67 boys operated on using conventional laparoscopy without lymphatic preservation between January 1997 and March 1999. Using 10x to 20x optical magnification, the lymphatic vessels were identified as colorless tubular structures that were easily separated and preserved. RESULTS: After a mean followup of 17 months there was no significant difference in varicocele recurrence between the 2 groups (6.7% vs 8.9%, p = 0.56). Hydrocele formation and testicular hypertrophy occurred significantly less frequently after microsurgical repair (1.9% and 2.9%, respectively, vs 17.9% and 20.1% in the conventional group, p = 0.0003). No major complications were encountered. CONCLUSIONS: Preservation of lymphatics in laparoscopic varicocelectomy is technically feasible, and decreases hydrocele formation and the development of testicular hypertrophy. This microsurgical modification is a safe and efficacious alternative for urologists skilled in reconstructive laparoscopy. PMID- 15821581 TI - Immunohistochemical markers in the diagnosis of prostate cancer. PMID- 15821582 TI - Adrenal myelolipoma. PMID- 15821583 TI - Over expression of hypoxia-inducible factor-1alpha in renal and bladder cancer cells increases tumorigenic potency. AB - PURPOSE: Hypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor that regulates genes involved in the response to hypoxia. We evaluated the effects of HIF-1alpha over expression on the tumorigenic potency of renal cell carcinoma VMRC cells and bladder cancer EJ cells in vitro and in vivo. MATERIALS AND METHODS: We introduced HIF-1alpha expression vectors into VMRC and EJ cells, and generated the HIF-1alpha over expressing cell lines VMRC-HIF1alpha and EJ HIF1alpha, and the vector only transfected cell lines VMRC-neo and EJ-neo. We then evaluated in vitro cell proliferation and in vivo tumor growth of these cell lines after subcutaneous injection into athymic nude mice. RESULTS: In vitro studies showed that HIF-1alpha over expression in VMRC and EJ cells accelerated cell proliferation during the confluent growth phase and rendered these cells resistant to hypoxic stress. Furthermore, in vivo studies revealed that all 4 types of cancer cells (VMRC-neo, VMRC-HIF1alpha, EJ-neo and EJ-HIF1alpha) formed tumors in nude mice and the size of VMRC-HIF1alpha cell derived xenografts was much larger than that of VMRC-neo cell derived xenografts. Although HIF-1alpha over expression did not affect the size of EJ cell derived xenografts, histological examination showed that there was only a small area of necrosis in EJ-HIF1alpha cell derived xenografts, whereas a large area of central necrosis was observed in EJ-neo cell derived xenografts. It was also found that HIF-1alpha over expression increased intratumor microvessel density in the xenografts. CONCLUSIONS: These results suggest that HIF-1alpha may have important roles in bladder and renal cancer angiogenesis and proliferation. PMID- 15821584 TI - DNA hypermethylation on multiple CpG islands associated with increased DNA methyltransferase DNMT1 protein expression during multistage urothelial carcinogenesis. AB - PURPOSE: We elucidated the significance of aberrant DNA methylation on multiple CpG islands and its correlation with DNA methyltransferase DNMT1 protein expression during urothelial carcinogenesis. MATERIALS AND METHODS: We examined the DNA methylation status on multiple CpG islands by methylation specific polymerase chain reaction and combined bisulfite restriction enzyme analysis in 12 specimens of normal urothelium, 23 of noncancerous urothelium showing no remarkable histological changes obtained from patients with bladder cancer (NBC) and 70 of transitional cell carcinoma (TCC). RESULTS: DNA methylation on CpG islands of the p16 (0%, 17% and 21%) and death-associated protein kinase (13%, 33% and 29%) genes, and methylated in tumor-2 (56%, 60% and 76%), 12 (0%, 6% and 30%), 25 (25%, 27% and 35%) and 31 (45%, 56% and 79%) clones was detected in normal urothelium, NBCs and TCCs, respectively. The incidence of concurrent DNA hypermethylation on 3 or more CpG islands in NBCs (38%) was significantly higher than that in normal urothelium (0%, p = 0.0455) and even higher in TCCs (59%, p = 0.0043). The incidence of the CpG island methylator phenotype in nonpapillary carcinomas (nodular invasive carcinomas and their precursors, ie flat carcinoma in situ, 71%) was significantly higher than in papillary carcinomas (40%, p = 0.0143). In all specimens examined concurrent DNA hypermethylation on 3 or more CpG islands significantly correlated with immunohistochemically evaluated DNMT1 protein over expression (p = 0.0167). CONCLUSIONS: DNA hypermethylation on multiple CpG islands in association with DNMT1 protein over expression may participate in multistage urothelial carcinogenesis even at the precancerous stage and particularly in the development of nodular invasive carcinomas of the bladder. PMID- 15821585 TI - Androgen mediated regulation and functional implications of fkbp51 expression in prostate cancer. AB - PURPOSE: Androgen ablation continues to be the most effective therapy for metastatic prostate cancer, although the biologically active androgen receptor (AR) target genes remain largely unknown. Because AR signaling continues in hormone refractory disease, effector AR target genes may have therapeutic import. MATERIALS AND METHODS: We used oligonucleotide microarrays to identify genes with expression induced by androgen and associated with androgen independent growth. The androgen induced expression of FKBP51, a steroid receptor chaperone, was further investigated in LNCaP cells by Northern and Western analysis, and in primary prostate specimens using immunohistochemistry. We used stable clones over expressing FKBP51 to test the functional effects of FKBP51. RESULTS: Many genes had expression that correlates with androgen stimulation in LNCaP cells but relatively few had reproducible, androgen mediated changes in expression across multiple prostate cancer cell lines. FKBP51 had androgen induced RNA and protein expression in LNCaP cells and decreased expression in normal prostate epithelial cells following castration. Further study demonstrated that FKBP51 induction was not a generalized response to cell proliferation, FKBP51 protein physically interacts with AR and LNCaP cells constitutively over expressing FKBP51 have increased ligand mediated AR activation of an exogenous AR reporter construct and endogenous prostate specific antigen. CONCLUSIONS: Taken together these results confirm FKBP51 as an androgen induced gene, demonstrate a physical interaction between FKBP51 and AR and suggest that FKBP51 over expression increases AR transcriptional activity in prostate cancer. PMID- 15821586 TI - Increased expression of MUC18 correlates with the metastatic progression of mouse prostate adenocarcinoma in the TRAMP model. AB - PURPOSE: The transgenic adenocarcinoma mouse prostate (TRAMP) model is a paradigm that closely mimics the progression of clinical prostate cancer. We have previously reported that MUC18, a cell adhesion molecule in the Ig gene superfamily, is a marker as well as an important mediator for the metastatic potential of human prostate cancer cells. In this study we investigated the possible correlation of increased MUC18 expression with the malignant progression of prostate cancer in the TRAMP model. MATERIALS AND METHODS: We used immunohistochemistry, Western blot and reverse transcriptase-polymerase chain reaction analyses to determine MUC18 expression in the prostate gland of 178 to 282-day-old TRAMP positive males with a prostate tumor size of 0.4 to 12.7 gm. Eight normal prostates, 10 prostates with high grade prostatic intraepithelial neoplasia (PIN), 24 prostates with primary prostate cancer, 10 metastatic lesions from 50 pure C57BL/6 TRAMP mice (Wu colony) and 2 normal prostates, 2 prostates with high grade PIN, 6 prostates with primary prostate cancer and 4 metastatic lesions from 10 [C57BL/6 TRAMP x FVB] F1 mice (NMG colony) were used. RESULTS: We found that mouse MUC18 was expressed in all (100%) high grade PIN, adenocarcinomas and metastatic lesions. All mice bearing primary prostate tumors had prostate cancer metastatic to the peri-aortic lymph nodes and some had it to other organs (liver, lung, kidney, testes, seminal vesicles and abdominal cavity). In contrast, prostates from 10 nontransgenic littermates did not have detectable MUC18 expression. CONCLUSIONS: MUC18 expression is up-regulated in the TRAMP model and it correlates with the malignant progression of mouse prostate adenocarcinoma in this transgenic model. This further strengthens the hypothesis that MUC18 has an important role in increasing the metastatic potential of prostate cancer cells. PMID- 15821587 TI - Trypanosoma cruzi infection induced changes in the innervation, structure and function of the murine bladder. AB - PURPOSE: The involvement of the lower urinary tract in chronic Chagas' disease has received little attention. Therefore, we investigated pathology and functional alterations in the bladder of Trypanosoma cruzi infected mice. MATERIALS AND METHODS: CD1 mice were infected with 5 x 10 T. cruzi trypomastigotes of the Brazil strain of T. cruzi. At day 100 after infection bladder structure and function were examined by pathological evaluation, magnetic resonance imaging and cystometric studies. RESULTS: The bladder in infected mice weighed more and were large, dilated, deformed, friable and thin walled compared with control mice. Magnetic resonance imaging confirmed these observations. Inflammation, fibrosis and ganglionitis was observed. Cystometric studies revealed that baseline, threshold and micturition pressures were increased in infected mice. Bladder overactivity and decreased bladder compliance were also noted in infected mice. There were no detectable differences in bladder capacity, micturition volume or residual volume between infected and uninfected mice. CONCLUSIONS: Bladder abnormalities may be a more common clinical sequelae of T. cruzi infection than previously appreciated. PMID- 15821588 TI - Urinary glycosaminoglycan excretion during the menstrual cycle in normal young women. AB - PURPOSE: We investigated whether the menstrual cycle affects urinary glycosaminoglycan (GAG) excretion in normal young women. MATERIALS AND METHODS: Urine samples from 10 healthy women 19 to 21 years old were collected daily during the whole menstrual cycle. Concentration of total urinary GAG was assessed as mug hexuronic acid per mg creatinine. Proportions of sulfated GAG species were determined by agarose gel electrophoresis. RESULTS: Mean excretion values +/- SD for period days 4 to 13 and 15 to 28 of the cycle were significantly different (0.445 +/- 0.041 vs 0.356 +/- 0.035 microg/mg, p <0.001). Correlation between values for the first and second halves of the cycle showed that this difference was consistent irrespective of individual variations in GAG excretion (r = 0.9757, p <0.001). Proportions of urinary sulfated GAG did not change during the cycle. CONCLUSIONS: Excretion of total urinary GAG during the normal menstrual cycle of young women has a biphasic pattern with significantly higher values occurring in the first half of the cycle. This variation implies modulation by estrogens and consequently it should be considered when comparing the GAG concentration in urine samples from women of childbearing age. PMID- 15821589 TI - Peroxynitrite may be involved in bladder damage caused by cyclophosphamide in rats. AB - PURPOSE: It was previously shown that nitric oxide (NO) produced by inducible NO synthase (iNOS) is responsible for cyclophosphamide (CP) induced cystitis. In this study we evaluated whether peroxynitrite is also responsible for CP induced bladder damage in rats. MATERIALS AND METHODS: A total of 38 male albino Wistar rats were divided into 4 groups. Group 1 served as controls and was given 2 ml saline, while 3 groups received a single dose of CP (200 mg/kg) at the same intervals. Group 2 received CP only, group 3 received the selective iNOS inhibitor aminoguanidine (AG) (100 mg/kg) and group 4 received the peroxynitrite scavenger ebselen (2-phenyl-1,2-benzisoselenazol-3[2H]-one) (20 mg/kg). RESULTS: CP injection resulted in severe cystitis with continuous macroscopic hemorrhage, strong edema, inflammation and ulceration. Moreover, bladder tissue malondialdehyde levels, iNOS activation and urine nitrite-nitrate levels were dramatically increased. AG histologically protected bladder against CP damage and decreased urine nitrite-nitrate levels, bladder malondialdehyde and iNOS induction. Ebselen showed results similar to those of AG without changing the urinary nitrite-nitrate level and iNOS activity. CONCLUSIONS: These results suggest that not only nitric oxide, but also peroxynitrite may be important in the pathogenesis of CP induced cystitis. PMID- 15821590 TI - In vitro comparison of stone retropulsion and fragmentation of the frequency doubled, double pulse nd:yag laser and the holmium:yag laser. AB - PURPOSE: The frequency doubled, double pulse Nd:YAG (FREDDY) laser (World of Medicine, Berlin, Germany) functions through the generation of a plasma bubble. Upon bubble collapse a mechanical shock wave is generated, causing stone fragmentation. This mechanism of action is in contrast to the holmium laser, which cause stone destruction by vaporization. Observed clinical stone retropulsion and fragmentation with the FREDDY and holmium lasers has prompted a series of in vitro experiments designed to compare laser induced retropulsion and fragmentation with those of a holmium laser and pneumatic lithotrite. MATERIALS AND METHODS: For retropulsion a hands-off underwater laboratory setup, including a horizontally oriented silicone tube 1.3 cm in diameter and a holder to keep the stone phantom in contact with the quartz laser fiber or pneumatic probe, was used. Previously weighed, cylindrical Bego stone phantoms (Bego USA, Smithfield, Rhode Island) were placed in the apparatus. Stone fragmentation was performed with the FREDDY or holmium laser, or the pneumatic lithotripter. The FREDDY and holmium lasers were tested at similar pulse energy and frequency settings. As a standard for comparison, a pneumatic lithotrite was tested with a semirigid probe and single pulse settings of 100, 200 and 300 kPa. Stone phantoms underwent 30 shocks per setting. Mean net retropulsion, defined as the final resting point of the stone, as determined by direct measurement, was recorded for each setting. For fragmentation plaster of Paris stone phantoms of known weights were used to compare the fragmentation ability of each laser. Stones phantoms were placed in a hands-off underwater setup, consisting of an inverted silicon syringe and holder immersed in tap water. The laser fiber (365 microm for the holmium and 280 microm for the FREDDY) was placed through the tip of the syringe in contact with the stone phantom. A total of 24 stones were divided into 4 groups of 6 per group. Two groups were fragmented with the FREDDY laser at 300 and 400 J total energy. The other 2 groups were fragmented using the holmium laser at 300 and 480 J total energy. Fragmentation efficiency was determined as percent weight loss. RESULTS: For retropulsion at 160 mJ the FREDDY laser caused stone retropulsion to a mean distance of 7.6, 8.1 and 6.8 cm at settings of 5, 10 and 15 Hz, respectively. At 0.8 J the holmium laser retropulsed the stone to a mean distance of 3.3 and 4.9 cm at settings of 5 and 10 Hz, respectively. The pneumatic device caused stone retropulsion a mean distance of 8.5, 9.9 and 13.8 cm at pressure settings of 100, 200 and 300 kPa, respectively. The FREDDY laser generally caused less retropulsion than the pneumatic device, although this difference was only significant at the highest pneumatic lithoclast setting (p <0.05). At clinically relevant settings the FREDDY laser caused significantly more retropulsion than the holmium laser (p <0.05). For fragmentation at total energy settings of 300 and 400 J the FREDDY laser resulted in 44.9% and 86.8% weight loss, respectively (p <0.05). At settings of 300 and 480 J the holmium:YAG laser resulted in 3.3% and 7.1% weight loss, respectively (p <0.05). CONCLUSIONS: At lower frequency settings stone retropulsion was significantly greater with the FREDDY laser compared with the holmium laser. However, retropulsion was significantly less than that caused by the pneumatic lithotripter at all settings. Therefore, we recommend the use of an occlusive device, such as the Stone Cone (Boston Scientific, Natick, Massachusetts) proximal to the calculus during intracorporeal ureteral lithotripsy and in the ureteropelvic junction during percutaneous laser nephrostolithotomy. In vitro stone fragmentation was significantly greater with the FREDDY laser than with the holmium:YAG laser, suggesting that the FREDDY may offer a low cost alternative to the holmium:YAG laser lithotrite in select patients. PMID- 15821591 TI - Gelatin matrix treatment of complex renal injuries in a porcine model. AB - PURPOSE: We developed a complex porcine renal laceration model at our laboratory to test the efficacy of an abbreviated, sutureless treatment regimen using FloSeal gelatin matrix hemostatic sealant (Baxter Healthcare Corp., Fremont, California). We evaluated the influence of preliminary renal arterial occlusion on hemostatic efficacy and assessed the risk of delayed hematoma or urinoma formation after treatment. MATERIALS AND METHODS: A total of 21 commercial swine underwent celiotomy with creation of a complex upper pole renal injury using a 4 x 4 cm cruciate press instrument. The injury was uniformly produced at the caudal extent of the upper third portion of the left kidney. Subjects were prospectively randomized into 3 treatment groups. Experimental animals were treated with the application of gelatin matrix with (7 in group 1) or without (7 in group 2) preliminary renal arterial occlusion. Control subjects (7 in group 3) were treated with conventional horizontal mattress sutured gelatin sponge bolsters over the capsular injury. Operative blood loss, time to hemostasis and volume of gelatin matrix required for hemostasis were compared among the groups. Abdominal computerized tomography with intravenous contrast medium was performed in each animal 7 days postoperatively. RESULTS: Gelatin matrix use resulted in significantly less mean blood loss (80.7 and 99.0 ml in groups 1 and 2, respectively) vs conventional suture treatment (191.8 ml in group 3, p = 0.036). Time to hemostasis was similarly decreased (1.1, 2 to 2.5 and 5.8 minutes in groups 1 to 3, respectively, p = 0.009). Followup abdominal computerized tomography with contrast medium revealed no clinically significant perinephric fluid collections (greater than 2 cm). CONCLUSIONS: FloSeal gelatin matrix hemostatic sealant provided effective hemostasis after complex renal injury with and without preliminary vascular control. No delayed bleeding or clinically significant urinoma formation was noted. These findings suggest a possible increased role for FloSeal gelatin matrix in renal salvage surgery. PMID- 15821592 TI - A method for producing overactive bladder in the rat and investigation of the effects of GABAergic receptor agonists and glutamatergic receptor antagonists on the cystometrogram. AB - PURPOSE: We induced radio frequency (RF) lesions in the neuronal pathway leading from the forebrain to the pontine micturition center (PMC) to produce a rat model of bladder overactivity. We studied the effects of gamma-aminobutyric acid agonists (diazepam and baclofen) and glutamate receptor antagonists (MK-801 maleate and GYKI52466 [1-(4-aminophenyl-D-4-methyl-7,8 methylenedioxy-5H-2,3 benzodiazepine] hydrochloride) on the cystometrogram and developed a possible explanation of the neuronal mechanisms underlying RF lesion induced bladder overactivity. MATERIALS AND METHOD: Seven-week-old male Sprague-Dawley rats were anesthetized with sodium pentobarbital and RF lesions were produced in the nuclei basalis. Five days later bladder contractions were induced by infusing fluid into the bladder and cystometrograms were measured in conscious rats. RESULTS: The micturition interval (MI) in rats subjected to RF lesioning was significantly shorter than that in sham operated control rats. Diazepam (0.1 and 1 mg/kg intraperitoneally), baclofen (1 mg/kg intravenously) and MK-801 (0.1 and 1 mg/kg intravenously) did not change or shortened MI in control rats but it prolonged MI in lesioned rats. GYKI52466 (0.5 and 1 mg/kg intravenously) weakly prolonged MI in lesioned rats. CONCLUSIONS: We consider that RF lesioning causes interruption of the inhibitory GABAergic neurons that lead from the forebrain to the PMC. This results in the activation of N-methyl-D-aspartate receptors in the PMC that are involved in the facilitation of voiding. PMID- 15821593 TI - Activation of the rostral pontine reticular formation increases the spinal glycine level and inhibits bladder contraction in rats. AB - PURPOSE: We examined the mechanism involved in the inhibition of bladder activity in rats by stimulating the rostral pontine reticular formation (RPRF) using carbachol, flavoxate and propiverine, and by analysis of amino acid levels in the lumbosacral cord. MATERIALS AND METHODS: A total of 82 female rats were anesthetized with urethane. Under isovolumetric conditions physiological saline, carbachol, flavoxate or propiverine was injected into the RPRF or intravenously. Changes in bladder activity and amino acid levels in the lumbosacral cord were examined. RESULTS: Injection of carbachol or flavoxate (0.3 microM each) into the RPRF abolished bladder contraction but there was no change after injection of physiological saline or propiverine. Intravenous injection of flavoxate or propiverine (0.1 to 10 mg/kg each) inhibited bladder contraction. Amino acid analysis revealed that injection of carbachol into the RPRF increased glutamate and glycine levels in the lumbosacral cord, while injection of flavoxate into the RPRF or intravenously caused an increase in glycine the lumbosacral cord. Injection of propiverine into the RPRF or intravenously did not influence lumbosacral cord amino acid levels. CONCLUSIONS: These results suggest that the RPRF has an important role in the inhibition of bladder contraction and carbachol or flavoxate can activate descending RPRF neurons and inhibit bladder contraction via spinal glycinergic neurons. PMID- 15821594 TI - Long-term efficacy of a vaginal sling procedure in a rat model of stress urinary incontinence. AB - PURPOSE: We examined the long-term efficacy of a newly created vaginal sling procedure for the restoration of leak point pressure (LPP) in a rat model of stress urinary incontinence (SUI). MATERIAL AND METHODS: A total of 20 female Sprague-Dawley rats were randomly assigned to 1 of 4 groups, namely normal control, SUI plus vaginal sling, SUI plus sham sling and SUI only. SUI was created in the latter 3 groups by bilateral pudendal nerve transection (PNT). In the sling procedure a 2 x 0.3 cm strip of polypropylene mesh was placed at the mid urethral level. Animals in the SUI plus sham sling group underwent vaginal dissection only. After 5 weeks LPP was measured 4 or 5 times in each rat and the mean was determined. The Kruskal-Wallis and Wilcoxon rank sum tests were used to evaluate whether levels of measurements differed across and between groups. RESULTS: Mean LPP +/- SD in control rats was 48.8 +/- 10.2 cm H2O. PNT decreased LPP to 23.5 +/- 7.4 cm H2O. Sling placement improved LPP at 5 weeks to 35.5 +/- 2.3 cm H2O, whereas LPP in the sham sling group was 29.1 +/- 4.9 cm H2O. LPP recovery in the sling group was significantly above levels for PNT (p = 0.037). LPP in the sling group did not differ statistically from that in the control group (p = 0.11). CONCLUSIONS: The newly created vaginal sling model restores the LPP in the rat model of SUI in the long term (5 weeks). This model could be used to address research questions related to the sling procedure. PMID- 15821595 TI - Improvement in erectile dysfunction after neurotrophic factor gene therapy in diabetic rats. AB - PURPOSE: Erectile dysfunction (ED) is a common but difficult to treat complication of diabetes mellitus (DM). We have previously reported herpes simplex virus (HSV) vector mediated delivery of nerve growth factor into the bladder to treat diabetic cystopathy and neurotrophin-3 (NT3) gene transfer for pyridoxine treatment. Nerve growth factor and NT3 are neurotrophic factors that may protect nerves from mechanical and metabolic damage. We investigated the effects of HSV mediated delivery of NT3 for the treatment of diabetic ED. MATERIALS AND METHODS: Male Sprague-Dawley rats weighing 300 to 400 gm were injected with 65 mg/kg streptozotocin to induce DM. After 4 weeks 20 microl containing 5 x 10 pfu replication defective HSV vector expressing lacZ (6 rats) or NT3 (6) were injected directly into the cavernous nerve sheath with a 30 gauge needle. Four weeks later the animals underwent measurement of intracavernous pressure under electrical stimulation (20 Hz, 0.5 millisecond and 10 V) of the cavernous nerve. Staining for lacZ and neuronal nitric oxide synthase in the major pelvic ganglia was also performed. RESULTS: beta-Galactosidase staining revealed lacZ positive neurons in the major pelvic ganglia. Maximal intracavernous pressure induced by electrical stimulation showed statistically significant mean values +/- SEM of 15.1 +/- 2.1 and 43.8 +/- 11.1 cm H2O in the lacZ and NT3 vector injected groups, respectively (p = 0.03). The mean number of neuronal nitric oxide synthase positive neurons per section in the NT3 group was significantly higher than that in the lacZ control group at 3.33 +/- 0.23 and 0.64 +/- 0.14 neurons per high power field, respectively (p < 0.001). CONCLUSIONS: We report that gene therapy for the treatment of diabetic ED is feasible with HSV vectors. NT3 gene therapy may be applicable for the treatment of ED induced by DM. PMID- 15821599 TI - Re: impact of surgical delay on long-term cancer control for clinically localized prostate cancer. PMID- 15821600 TI - Re: laparoscopic assisted radical cystectomy with ileal neobladder: a comparison with the open approach. PMID- 15821601 TI - Re: an evaluation of the decreasing incidence of positive surgical margins in a large retropubic prostatectomy series. PMID- 15821604 TI - Re: osteoporosis and spinal fractures in men with prostate cancer: risk factors and effects of androgen deprivation therapy. PMID- 15821605 TI - Re: osteoporosis and spinal fractures in men with prostate cancer: risk factors and effects of androgen deprivation therapy. PMID- 15821606 TI - Re: midline dorsal plication technique for penile curvature repair. PMID- 15821607 TI - Re: comparison of guidewires in urology. Which, when and why? PMID- 15821608 TI - Re: transdermal application of verapamil gel to the penile shaft fails to infiltrate the tunica albuginea. PMID- 15821610 TI - Re: secondary malignancies in different forms of urinary diversion using isolated gut. PMID- 15821615 TI - Preoperative imaging of surrounding arteries by three-dimensional CT: is it useful for laparoscopic gastrectomy? AB - One of the difficulties in laparoscopic gastrectomy is the identification of the feeding artery of the stomach. Recently, 3-dimensional computed tomographic angiography has enabled the noninvasive visualization of arteries surrounding the stomach. Preoperative 3-dimensional computed tomographic angiography may facilitate laparoscopic gastrectomy by obtaining a road map of the arteries of the stomach. Twenty-nine cases of gastric cancer were evaluated using 3 dimensional computed tomographic angiography before surgery. Three-dimensional computed tomographic angiography showed the left gastric artery in 29 patients (100%), the right gastroepiploic artery in 29 patients (100%), the right gastric artery in 24 patients (82.8%), and the left gastroepiploic artery in 21 patients (72.4%). The aberrant hepatic artery was detected in 6 patients, and the variant of the right gastric artery and the variant of the left gastric artery were detected in 5 cases and 1 case, respectively. All laparoscopic gastrectomy procedures were performed successfully without conversion to open surgery. Preoperative 3-dimensional computed tomographic angiography was considered to be useful for laparoscopic gastrectomy. PMID- 15821616 TI - Laparoscopic fenestration of symptomatic liver cysts. AB - The purpose of this clinical study was to evaluate the usefulness of laparoscopic fenestration of symptomatic liver cysts. Between September 1996 and September 2001, 6 patients underwent laparoscopic fenestration for symptomatic hepatic cysts. All 6 patients were women. The mean age was 59.5 (range 40-74). Two patients had single and 4 had multiple cysts. The mean diameter of the lesions, measured by preoperative computed tomography (CT), was 16.8 cm (range 10-20). The indications for surgical treatment included abdominal fullness associated with pain or dyspepsia. The surgical procedure involved puncture and aspiration of the cyst and subsequent circular electroresection of the cystic wall. Laparoscopic fenestration was successfully done in all 6 patients. The operating time was 148 minutes (range 110-215). Mean blood loss was 93 mL (range 5-300). Histologic examination of the cyst wall showed 5 simple cysts and 1 papillary serous cystadenoma. There were no intraoperative or postoperative complications. The mean hospital stay after surgery was 4.3 days (range 3-5). Complete relief of symptoms was achieved in 5 patients during the postoperative follow-up (range 2-6 years, mean 4). Follow-up CT (1 month to 4 years) has shown regression of the cysts in all patients (mean 3.8 cm, range 2-8). Laparoscopic fenestration of symptomatic liver cysts is a simple and effective method to relieve symptoms with minimal surgical trauma. PMID- 15821617 TI - Safety of laparoscopic intracorporeal rectal transection with double-stapling technique anastomosis. AB - To assess the feasibility and analyze the short-term outcomes of laparoscopic intracorporeal rectal transection with double-stapling technique anastomosis, a review was performed of a prospective registry of 67 patients who underwent laparoscopic sigmoidectomy and anterior resection with intracorporeal rectal transection and double-stapling technique anastomosis between July 2001 and January 2004. Patients were divided into 3 groups: sigmoid colon/rectosigmoid carcinoma, upper rectal carcinoma, and middle/lower rectal carcinoma. A comparison was made of the short-term outcomes among the groups. The number of cartridges required in bowel transection was significantly increased in patients with middle/lower rectal carcinoma, and significant differences were observed in the length of the first stapler cartridge fired for rectal transection. Furthermore, mean operative time and blood loss were also significantly greater in the middle/lower rectum group; however, complication rates and postoperative course were similar among the 3 groups. No anastomotic leakage was observed. Laparoscopic intracorporeal rectal transection with double-stapling technique anastomosis can be performed safely without increased morbidity or mortality. PMID- 15821618 TI - Comparison of needlescopic appendectomy versus conventional laparoscopic appendectomy: a randomized controlled trial. AB - Laparoscopic appendectomy has been shown to improve postoperative recovery when compared with open appendectomy. The present randomized trial was conducted to evaluate any further difference in outcome between needlescopic appendectomy (NA) and conventional laparoscopic appendectomy (CLA) in the management of acute appendicitis. Patients with the clinical diagnosis of acute appendicitis were randomized to either NA (instrument size < or = 3 mm) or CLA (instrument size > or = 5 mm). Standardized anesthetic technique and perioperative management were adopted. The primary end point was length of postoperative hospital stay. Other parameters such as conversion rate, postoperative pain score and analgesic requirement, return of bowel function, resumption of normal activities, complication rate, and length of the final scars were also assessed and compared. A total of 363 patients (NA: 174, CLA: 189) were recruited. Both approaches could accurately arrive at the diagnosis (NA: 98.3%; CLA: 100%). Compared with CLA, NA resulted in a significantly longer operation time (P = 0.015) and a higher conversion rate (P < 0.001). The final scars of the NA group were significantly shorter when compared with the CLA group (P < 0.001). Otherwise, there was no statistical difference between the 2 groups in terms of complication rate, postoperative pain score, length of postoperative stay, and other recovery parameters. NA resulted in a longer operation time and higher conversion rate. Except for a smaller scar, the present study was unable to demonstrate any other short-term benefits. Thus, the technique cannot be routinely recommended. PMID- 15821619 TI - The safety and efficacy of direct trocar insertion with elevation of the rectus sheath instead of the skin for pneumoperitoneum. AB - Abdominal access in endoscopic surgery carries a finite risk of visceral injury. Bleeding, subcutaneous emphysema, gastrointestinal tract perforation, minor and major vascular injury, and intraperitoneal adhesions are the potential complications associated with abdominal access and creation of pneumoperitoneum. There are 4 basic techniques used to create pneumoperitoneum: blind Veress needle, direct trocar insertion, optical trocar insertion, and open laparoscopy. Veress needle and direct trocar insertion are blind techniques, and their use can result in severe visceral and vascular injuries. To prevent visceral and vascular injuries caused by the technique used for the creation of pneumoperitoneum, laparoscopic surgeons and gynecologists look for safe and effective laparoscopic access techniques. Direct trocar insertion without previous pneumoperitoneum was reported to be a safe alternative to Veress needle insertion. We carried out this study to compare the ease of use, safety, and efficacy of direct trocar insertion with elevation of the rectus sheath and blind insertion of the Veress needle in laparoscopic surgery. In 578 laparoscopic procedures, the patients were assigned to one of the following groups: blind insertion of the Veress needle (group 1, n = 301) and direct trocar insertion with elevation of the rectus sheath using 2 towel clips (group 2, n = 277). Total complication rates were 15.7% (n = 33) and 3.3% (n = 4) in groups 1 and 2, respectively (P < 0.05). Direct trocar insertion with elevation of the rectus sheath using 2 towel clips is an easy, safe, and effective technique. PMID- 15821620 TI - A review of surgical simulation with attention to validation methodology. AB - The use of simulation technology for teaching and evaluating surgical skills has gained considerable attention in recent years. This is driven by interest in quality of care, concerns over increasing operative complexity, constraints on the use of animal models, limited available patient material, medicolegal pressures, and fiscal mandates for cost-effective performance. Traditional mechanical models are yielding to techniques dependent on electronic technology, including virtual reality. Data to support the validity of simulation techniques for surgical training, assessment, and certification represent only a fraction of the literature available on the subject. Literature searches were conducted in MEDLINE and ERIC, covering the period from 1966 to the present. The electronic and bioengineering literature was not surveyed due to the extensive literature on technology development, distinct from assessment of context specific validity. The search results and the bibliographies of key review articles were examined to identify articles that contained original data, measured performance between cohorts, defined performance measures, and described a standard against which performance was compared. Most of the literature pertaining to simulation techniques for surgical training has been published within the past 5 years and consist of review, opinion, and feasibility articles. There is an emerging body of evidence to establish the validity of simulation techniques for assessing surgical skills. Further refinement of simulation techniques, identification of specific performance measures, longitudinal evaluations, and comparison to practice outcomes are still needed to establish the validity and the value of surgical simulation for teaching and assessing surgical skills prior to considering implementation for certification purposes. PMID- 15821621 TI - Predictors of duration of postoperative hospital stay in patients undergoing advanced laparoscopic surgery. AB - The expansion of the indications for laparoscopic surgery to include high-risk patient, acute and malignant pathology, and more complex procedures may prolong the hospital stay. Cox multiple stepwise regression analysis model was employed to determine independent predictors of prolonged postoperative hospital stay (more than 3 days) following advanced laparoscopic procedures among 10 variables. Some 130 patients had undergone advanced laparoscopic surgical procedures between November 2000 and August 2003. The median postoperative hospital stay was 3 days (interquartile range 2-5), and 81 patients (62.3%) were discharged within 3 days of surgery. The independent predictors of prolonged postoperative hospital stay were ASA score of 3 or 4 (odds ratio [OR] = 4.610, P = 0.0002) and preoperative hospital stay (OR = 0.151 per day, P = 0.001). Independent predictors of duration of preoperative hospital stay were emergency admission to hospital (OR = 9.516, 95% CI 5.770-13.261, P < 0.0001) and an underlying malignant pathology (OR = 7.948, 95% CI 3.623-12.273, P = 0.0004). Advanced laparoscopic surgery is associated with a short postoperative hospital stay in the majority of patients. Prolongation of the postoperative hospital stay (more than 3 days) may be expected if the patient had been in the hospital with an acute or malignant disease for more than 6 days prior to surgery and in patients with high comorbidity. The duration of surgery has no impact on the duration of the postoperative hospital stay. PMID- 15821622 TI - Upper gastrointestinal bleeding in a patient with a balloon bumper PEG feeding tube. PMID- 15821623 TI - Endoscopic complete removal of the histologically unconfirmed gastric cancer with massive invasion as a giant biopsy. AB - A 68-year-old man underwent upper GI endoscopy in a mass screening, which showed a suspicious tumor in the antral region of stomach. However, histologic examination of repeated endoscopic biopsy samples (15 samples) revealed no neoplastic change. Although all the examinations including endoscopy, endosonography, and barium study pointed to the diagnosis of gastric cancer, we did not have histologic evidence of malignancy. Hence, complete endoscopic removal of the lesion using a new method of endoscopic mucosal resection (EMR) was performed as a giant biopsy. Histologic assessment of the endoscopically resected specimen confirmed invasive adenocarcinoma of the stomach. He then underwent distal gastrectomy with regional lymphadenectomy. Endoscopic resection of the tumor in the digestive tract is also a very useful diagnostic modality of the lesion when malignancy cannot be revealed by endoscopic biopsy. PMID- 15821624 TI - Spontaneous resolution of massive right-sided pneumothorax occurring during laparoscopic cholecystectomy. AB - Pneumothorax is a rare but potentially serious complication that can occur during laparoscopic surgery. We describe a case of a spontaneous massive right-sided pneumothorax that occurred during laparoscopic cholecystectomy, presumably because of escape of intraperitoneal carbon dioxide under pressure into the pleural cavity through a congenital defect in the diaphragm. During the procedure, arterial oxygen saturation decreased and clinical examination revealed signs of a right-sided pneumothorax. This was confirmed on chest x-ray in the immediate postoperative period. Since the patient was clinically stable without any signs of respiratory distress, a conservative approach was adopted. The patient remained on close clinical observation and continuous monitoring of arterial hemoglobin oxygen saturation by pulse oximetry and repeat chest x-rays and had an uneventful recovery with complete resolution of the pneumothorax 3 hours after surgery and without the need for thoracic aspiration or tube thoracostomy. PMID- 15821625 TI - Laparoscopic cholecystectomy in the presence of lymphangiomyomatosis. AB - A 49-year-old woman with a history of generalized lymphangiomyomatosis or lymphangioleiomyomatosis (LAM), including angiomyolipomas of the liver and kidney and known LAM of the lung, presented with symptomatic cholelithiasis. She underwent an uneventful laparoscopic cholecystectomy and was discharged home the following day. Careful patient selection, preoperative stratification, and optimization of pulmonary status are all necessary prior to electing to operate on patients with known LAM. A CT scan of the abdomen is of value to delineate the anatomy of the angiomyolipomas and their proximity to the operative site. These preliminary precautions, combined with careful surgical technique, are essential for a successful outcome with minimal morbidity. PMID- 15821626 TI - Laparoscopic transabdominal preperitoneal hernioplasty of bilateral obturator hernia. AB - Obturator hernia is relatively rare and tends to occur in elderly, emaciated women with chronic diseases. Clinical presentations are frequently delayed and so preoperative diagnosis is difficult. Treatment is always surgical. We present a case of a 75-year-old woman with bilateral obturator hernia diagnosed by the physical examination and abdominopelvic computed tomography (CT) scan; she had no signs of bowel strangulation. We used a laparoscopic approach for correction. A transabdominal preperitoneal hernioplasty was done using a prosthetic patch of polypropylene mesh. The patient recovered very well after surgery. We suggest that a laparoscopic approach may be used as treatment, when a nonstrangulated obturator hernia is diagnosed preoperatively. PMID- 15821627 TI - Thoracoscopic sympathetic clamping in a patient with an azygos fissure. AB - We believe that an azygos fissure may predispose to bleeding during thoracoscopic surgery. An azygos fissure causes important morphologic changes in the superior mediastinum and thereby poses a risk of massive bleeding during thoracoscopic procedures. We report on a successful thoracoscopic procedure conducted in a patient with palmar hyperhidrosis and an azygos fissure and emphasize that the course of the thoracic sympathetic chain runs laterally along the base of the azygos fissure. PMID- 15821628 TI - Suture passer tip breakage during laparoscopic ventral hernia repair. AB - Laparoscopic ventral hernia repair generally employs a tacker and a suture passer to secure the mesh to the abdominal wall. We reviewed cases of Gore Suture Passer tip breakage during these procedures and their management. Surgeons performing laparoscopic ventral hernia repair were asked about encountered complications relating to the Gore Suture Passer instrument. Charts of the patients with significant alteration in the course of their procedure secondary to such complication were reviewed. Two cases of suture passer tip breakage were identified. One required fluoroscopy to localize and recover the tip, resulting in significant prolongation of the operation. The other required conversion to laparotomy with mesh removal; the tip of the Gore Suture Passer was found in the pelvis and the hernia was repaired with a Stoppa technique. The Gore Suture Passer tip may break during laparoscopic ventral hernia repair, which may significantly complicate the case. PMID- 15821629 TI - An alternative port for use in hand-assisted laparoscopic surgery: a design using a stoma ring and a glove. AB - Hand-assisted laparoscopic surgery (HALS) has widened the range of laparoscopic surgery. An appropriate port allowing the passage of the hand while maintaining pneumoperitoneum is mandatory for HALS to be performed. Commercially available ports are expensive and may not be universally available. An inexpensive 2 component port using the flange of a stoma appliance and a glove, which could be readily assembled at short notice, is described. PMID- 15821631 TI - Re: Extraperitoneal laparoscopic approach to Spigelian hernia combined with groin hernias. PMID- 15821632 TI - Zegerid--immediate-release omeprazole. AB - The FDA has approved marketing of Zegerid powder for oral suspension (Santarus), an immediate-release formulation of the proton-pump inhibitor (PPI) omeprazole (Prilosec, and others). All other oral PPIs are delayed-release, enteric-coated formulations designed to prevent degradation of the drug by gastric acid. Each 20 or 40-mg packet of Zegerid contains 1680 mg sodium bicarbonate, which protects the drug from gastric acid degradation. A dose of Zegerid contains 460 mg of sodium, which may be excessive for some patients. Zegerid is the first oral PPI to be approved by the FDA for reduction of risk of upper GI bleeding in critically ill patients. The drug may be useful for patients who are unable to swallow and have nasogastric (NG) tubes in place. Zegerid cost $70.00 for 14 days' treatment, compared to less than $10 for 14 tablets of Prilosec OTC. PMID- 15821633 TI - Menactra: a meningococcal conjugate vaccine. AB - Menactra is the first conjugated meningococcal vaccine approved in the US. It protects against meningococcal serogroups A, C, Y and W-135. The CDC's Advisory Committee on Immunization Practices (ACIP) recommends administration of the new vaccine to pre-adolescents aged 11 or 12, teens entering high school and college freshmen living in dormitories. These broad recommendations have led to a shortage of the new vaccine. Menomune, the old unconjugated quadrivalent meningococcal vaccine, is also effective in these age groups and could still be used. PMID- 15821634 TI - Uterine artery embolization for fibroids. AB - UAE is an alternative to surgery for treatment of uterine fibroids that uses injections of small plastic or gelatin beads to thrombose fibroid vessels arising from the uterine artery. Clinical studies have reported less menorrhagia and pelvic pain in patients who have undergone the procedure. After embolization, many patients experience pain requiring an overnight hospital stay and administration of parenteral opioids. UAE is generally not recommended for women who have not completed childbearing. PMID- 15821635 TI - Drugs for asthma. AB - Patients with mild, infrequent asthma symptoms may require only intermittent, as needed use of an inhaled short-acting beta2-adrenergic agonist. Use of a short acting beta2-agonist more than twice weekly, other than for exercise-induced bronchospasm, indicates a need for anti-inflammatory treatment. Inhaled corticosteroids are the most effective anti-inflammatory medication; leukotriene modifiers are less effective alternatives. If regular use of an inhaled corticosteroid in a low dose does not prevent symptoms, a long-acting beta2 agonist should be added; addition of a second drug is more effective than raising the dose of the inhaled steroid. A leukotriene modifier can also be used as the second drug. Omalizumab may be considered as adjunctive therapy for patients more than 12 years old who have allergic asthma not controlled by other drugs. A short course of oral corticosteroids may be useful for acute exacerbations. Treatment of acute severe asthma as a medical emergency is not included here; it has been reviewed elsewhere (ER McFadden Jr, Am J Respir Crit Care Med 2003; 168:740). PMID- 15821636 TI - Surgical intervention for T4 lung cancer with infiltration of the thoracic aorta: are we back to the archetype of surgical thinking? PMID- 15821637 TI - Diagnostic power of aortic elastic properties in young patients with Marfan syndrome. AB - BACKGROUND: In patients with Marfan syndrome, progressive aortic dilation implicates a still-unpredictable risk of life-threatening aortic dissection and rupture. We sought to quantify aortic wall dysfunction noninvasively, determine the diagnostic power of various aortic parameters, and establish a diagnostic model for the early detection of aortic abnormalities associated with Marfan syndrome. METHODS: In 19 patients with Marfan syndrome (age, 17.7 +/- 9.5 years) and 19 age- and sex-matched healthy control subjects, computerized ascending and abdominal aortic wall contour analysis with continuous determination of aortic diameters was performed out of transthoracic M-mode echocardiographic tracings. After simultaneous oscillometric blood pressure measurement, aortic elastic properties were determined automatically. RESULTS: The following ascending aortic elastic parameters showed statistically significant differences between the Marfan group and the control group: (1) decreased aortic distensibility ( P < .001), (2) increased wall stiffness index ( P < .01), (3) decreased systolic diameter increase ( P < .01), and (4) decreased maximum systolic area increase ( P < .001). The diagnostic power of all investigated parameters was tested by single logistic regression models. A multiple logistic regression model including solely aortic parameters yielded a sensitivity of 95% and a specificity of 100%. CONCLUSIONS: In young patients with Marfan syndrome, a computerized image analyzing technique revealed decreased aortic elastic properties expressed by parameters showing high diagnostic power. A multiple logistic regression model including merely aortic parameters can serve as useful predictor for Marfan syndrome. PMID- 15821638 TI - The effect of gene transfer with hepatocyte growth factor for pulmonary vascular hypoplasia in neonatal porcine model. AB - BACKGROUND: Severe degree of pulmonary vascular hypoplasia remains a major limitation in congenital heart surgery. Considering the potential effect of gene transfer with hepatocyte growth factor to induce the angiogenesis in the lung, we assessed the effects of hepatocyte growth factor gene transfer in neonatal porcine lung with pulmonary vascular hypoplasia to achieve treatment for severe pulmonary vascular hypoplasia. METHODS: The model of pulmonary vascular hypoplasia was introduced with left pulmonary artery banding in piglet lung. After 7 days of pulmonary artery banding, piglets were transfected selectively to the left lung via the left pulmonary artery with a hemagglutinating virus of Japan E vector bearing the cDNA encoding human hepatocyte growth factor (H group) or control vector (C group). RESULTS: Seven days after the transfection, selective angiography of the left pulmonary artery showed the progression of left pulmonary vascular hypoplasia of the left lung in the C group but a significant attenuation of left pulmonary vascular hypoplasia in the H group. A right pulmonary artery occlusion test showed a marked increase in right ventricular systolic pressure in the C group, but this was significantly attenuated in the H group (C: 22.0 +/- 2.9, H: 13.0 +/- 2.7 mm Hg; P < .05). Histologic examination revealed that hepatocyte growth factor gene transfection increased the pulmonary vasculature in the left lung. CONCLUSIONS: Our results demonstrated that gene transfer of hepatocyte growth factor via the pulmonary artery showed the angiogenic effects in porcine model of pulmonary vascular hypoplasia after pulmonary artery banding. PMID- 15821639 TI - Three-dimensional echocardiography improves the understanding of left atrioventricular valve morphology and function in atrioventricular septal defects undergoing patch augmentation. AB - OBJECTIVES: We sought to address the role of 3-dimensional echocardiography in the evaluation of the left atrioventricular valve in children with an atrioventricular septal defect who underwent patch augmentation of their valve for either regurgitation or left ventricular outflow tract obstruction. METHODS: Five children whose ages ranged between 4.5 and 9.2 years and who underwent patch augmentation of their left atrioventricular valve had a preoperative and postoperative transesophageal echocardiogram with 3-dimensional reconstruction to evaluate the left atrioventricular valve. The indication for operation was left atrioventricular valve regurgitation in 3 patients and left ventricular outflow tract obstruction in 2 patients. Three were rerepairs, and 2 were primary repairs. Both 3-dimensional morphology and color Doppler data were obtained. Two- and 3-dimensional findings were correlated with surgical observations through the use of direct inspection and video images obtained with a head-mounted super-VHS camera. RESULTS: In each case there was precise correlation between the 3 dimensional and surgical findings as to the cause of leaflet failure in those with regurgitation. The site that would require leaflet augmentation could be determined by means of 3-dimensional echocardiography. Three-dimensional echocardiography provided more specific detail as to the morphology and function of the left atrioventricular valve than did its 2-dimensional counterpart. CONCLUSIONS: Three-dimensional echocardiography provides detailed information about the status of the left atrioventricular valve in the atrioventricular septal defect and can aid in the planning of either primary or secondary repair. PMID- 15821640 TI - The effect of surgical case volume on outcome after the Norwood procedure. AB - OBJECTIVES: We analyze the effect of surgical case volume on the survival of infants with hypoplastic left heart syndrome who underwent stage I surgical palliation (the Norwood procedure). The purpose of our study was to understand more clearly the relative effects of institution and surgeon experience on patient outcome. METHODS: Using the Pediatric Health Information System database belonging to the pediatric hospital members of the Child Health Corporation of America, we identified newborn infants (< 30 days old on admission) from 1998 through 2001 admitted with the diagnosis of hypoplastic left heart syndrome. Stepwise multiple regression analysis was used to examine the association between both institutional and surgeon case volume with 28-day survival after the Norwood procedure. RESULTS: Twenty-nine hospitals and 87 surgeons performed 801 Norwood procedures during the study period. In the 4 of 29 institutions that averaged 1 or more Norwood procedures per month during the study period, survival averaged 78%. The remaining 25 institutions averaged 1 Norwood procedure every 9.6 weeks, with a survival of 59%. Data analysis revealed that higher institutional volume (P = .02) but not the number of cases performed by surgeons (P = .13) increased survival after the Norwood procedure. There was no such association with average length of stay in survivors or the time to mortality in nonsurvivors. CONCLUSION: Survival after the Norwood procedure is associated with institutional Norwood procedure volume but not with individual surgeon case volume, suggesting the need for improvements in institutional-based approaches to the care of children with hypoplastic left heart syndrome and the need for establishing regional referral centers for such high-risk procedures to improve patient survival. PMID- 15821641 TI - Surgical trauma affects the proinflammatory status after cardiac surgery to a higher degree than cardiopulmonary bypass. AB - OBJECTIVES: Cytokines contribute to the development of the systemic inflammatory response syndrome or multiple-organ failure frequently observed after cardiopulmonary bypass-supported cardiac surgery. To quantify the contribution of bypass-induced versus trauma-induced inflammatory response after coronary artery bypass grafting, we examined plasma cytokine levels in 120 patients with coronary artery disease who were treated with or without cardiopulmonary bypass-assisted procedures. METHODS: Patients were treated in accordance with one of the following protocols: (1) elective percutaneous coronary intervention without cardiopulmonary bypass (n = 69), (2) cardiopulmonary bypass-supported percutaneous coronary intervention (cardiopulmonary bypass-percutaneous coronary intervention; n = 10), and (3) cardiopulmonary bypass-supported coronary artery bypass grafting (cardiopulmonary bypass-coronary artery bypass grafting; n = 41). Cytokine levels (picograms/milliliter) were measured by enzyme-linked immunosorbent assay from plasma samples obtained at various time points. RESULTS: Interleukin-6 was measured in blood samples from all 3 patient populations. The maximum interleukin-6 level was 13.6 +/- 22.3 pg/mL in the percutaneous coronary intervention group, 170.4 +/- 165.4 pg/mL in the cardiopulmonary bypass percutaneous coronary intervention group, and 640.3 +/- 285.7 pg/mL in the cardiopulmonary bypass-coronary artery bypass grafting group. Interleukin-6 levels were significantly different, and the 95% confidence intervals did not overlap. In the cardiopulmonary bypass-percutaneous coronary intervention group, bypass duration correlated well with interleukin-6 production ( r = 0.915; P < .001), whereas these parameters did not correlate in patients who underwent cardiopulmonary bypass-coronary artery bypass grafting ( r = 0.307; P = .054). CONCLUSIONS: These findings support the suggestion that surgical trauma and cardiopulmonary bypass contribute to the inflammatory response after cardiac surgery, although trauma may contribute to a higher degree. PMID- 15821642 TI - Experimental hypercholesterolemia differentially affects adventitial vasa vasorum and vessel structure of the left internal thoracic and coronary arteries. AB - OBJECTIVE: Atherosclerosis is a chronic and diffuse disease that affects all vascular beds. However, some vascular beds are more prone to atherosclerosis than others. Recent evidence suggests a role for the vasa vasorum in the atherosclerotic process. We hypothesized that there is a difference in adventitial vasa vasorum structure between the left internal thoracic artery and the coronary artery. Hence the current study was designed to characterize and compare the structure of the adventitial vasa vasorum in the left internal thoracic and coronary arteries. METHODS: Samples of vessels were obtained from female crossbred domestic pigs maintained on a normal (n = 6) or high-cholesterol (n = 6) diet for 12 weeks. The samples were scanned with micro-computed tomography, and the tomographic images were reconstructed and analyzed to obtain lumen area, vessel wall area, vasa vasorum count, vasa vasorum density, mean diameter of first- and second-order vasa vasorum, and second-order/first-order vasa vasorum ratio. RESULTS: Vasa vasorum density was significantly higher in the coronary arteries versus that seen in the left internal thoracic arteries in the normal group, as well as in the high-cholesterol group. The higher vasa vasorum density in the high-cholesterol group versus that in the normal group was significant for both vessels, being more pronounced in the left internal thoracic artery. Lumen area and second-order/first-order vasa vasorum ratio were higher in the high-cholesterol group than in the normal group only in the left internal thoracic artery. CONCLUSION: This study demonstrated that low vasa vasorum spatial density and higher lumen area observed in the left internal thoracic artery compared with that seen in the native coronary artery can be the structural background for the low incidence of atherosclerosis in this vessel. PMID- 15821643 TI - Osteopontin expression and adventitial angiogenesis induced by local vascular endothelial growth factor 165 reduces experimental aortic calcification. AB - BACKGROUND: Vascular calcification is a common pathologic and precisely regulated process involving bone-associated proteins such as osteopontin. In this study, we investigated mechanisms by which recombinant human vascular endothelial growth factor 165 protects the arterial wall from severe vascular remodeling, including calcification, a newly discovered biologic action of vascular endothelial growth factor. METHODS: In a rabbit model of thoracic aortic end-to-end anastomosis that simulates cardiovascular intervention, recombinant human vascular endothelial growth factor 165 at a dose of 0.75 mug (n = 19) or albumin (control; n = 19) was delivered intraluminally and on the serosal surface. Animals were killed, and aortic tissue was evaluated by Western blotting, immunohistochemistry, and immunofluorescence at 4, 8, and 24 hours; 1 week; and 1 month after surgery. RESULTS: All controls revealed extensive aortic medial calcification at 1 month, whereas calcification was significantly reduced or absent with vascular endothelial growth factor treatment. Compared with controls, vascular endothelial growth factor treatment resulted in an earlier infiltration of macrophages in the vessel media (at 8 hours: 5.7 +/- 2.3 macrophages per high-power field in control vs 32.1 +/- 7.5 in vascular endothelial growth factor-treated aortas; P < .001), whereas controls showed an increase in macrophages starting at 1 week (24.1 +/- 6.9 vs 4.3 +/- 1.8; P < .001). Osteopontin expression was transiently increased and detected in macrophages and endothelial cells in vascular endothelial growth factor-treated vessels, and adventitial microvascular density was significantly increased by 1 week (9.5 +/- 0.43 vs 25.0 +/- 1.3; P < .001). CONCLUSIONS: Our data suggest that exogenous vascular endothelial growth factor is capable of increasing adventitial angiogenesis and shifting macrophage infiltration and osteopontin expression in the media to an earlier time point, thereby promoting prompt repair and diminishing vascular remodeling and calcification after acute vascular injury. PMID- 15821644 TI - Combined administration of nitric oxide gas and iloprost during cardiopulmonary bypass reduces platelet dysfunction: a pilot clinical study. AB - BACKGROUND: Thrombocytopenia and platelet dysfunction are major mechanisms of cardiopulmonary bypass-induced postoperative hemorrhage. This study evaluated the effects of low amounts of nitric oxide, iloprost (prostacyclin analog), and their combination administered directly into the oxygenator on platelet function, platelet-leukocyte interactions, and postoperative blood loss in patients undergoing coronary artery bypass grafting. METHODS: Blood samples from 41 patients randomized to the control, nitric oxide (20 ppm), iloprost (2 ng x kg -1 x min -1 ), or nitric oxide plus iloprost groups were collected during cardiopulmonary bypass. Platelets and leukocytes were enumerated. Platelet membrane glycoprotein Ib and glycoprotein IIb/IIIa, P-selectin, platelet-derived microparticles, leukocyte CD11b/CD18 (Mac-1), and platelet-leukocyte aggregate were quantified by means of flow cytometry. Collagen and thrombin receptor activating peptide-induced platelet aggregation in whole blood was analyzed by means of aggregometry. RESULTS: Both nitric oxide or iloprost attenuated cardiopulmonary bypass-induced thrombocytopenia, reduction of glycoprotein Ib and glycoprotein IIb levels, translocation of P-selectin, microparticle formation, Mac-1 upregulation, and suppression of collagen-induced aggregation. Nitric oxide plus iloprost was significantly more effective in preventing thrombocytopenia, microparticle formation, and P-selectin translocation. Moreover, this treatment preserved thrombin receptor-activating peptide-induced aggregation, which was not rescued by single treatments. Both nitric oxide and nitric oxide plus iloprost attenuated postoperative blood loss. CONCLUSIONS: Nitric oxide plus iloprost reduced the deleterious effects of cardiopulmonary bypass, such as thrombocytopenia, platelet activation, platelet-leukocyte aggregate formation, and suppression of platelet aggregative responses. The reduced postoperative bleeding observed with this treatment suggests that this is a new and clinically feasible therapeutic option for patients subjected to cardiopulmonary bypass. PMID- 15821645 TI - Alterations in transmural strains adjacent to ischemic myocardium during acute midcircumflex occlusion. AB - OBJECTIVE: Helically oriented left ventricular fibers assemble into transmural sheets, which are important for wall-thickening mechanics: 15% fiber shortening results in 40% cross-fiber left ventricular wall thickening and a 60% ejection fraction through sheet extension, thickening, and shear. Normal cardiac microstructure and strains are optimized; deviations could result in apoptosis and deleterious matrix remodeling, which degenerates into global cardiomyopathy. We studied alterations in transmural strains adjacent to ischemic myocardium during acute midcircumflex occlusion. METHODS: Nine sheep had radiopaque markers implanted to measure left ventricular systolic fractional area shortening; 3 transmural bead columns were inserted into the midlateral wall for strain analysis. Three-dimensional marker coordinates were obtained with biplane videofluoroscopy before and during 70 seconds of ischemia. Systolic strains were quantified along circumferential, longitudinal, and radial axes (n = 9) and were transformed into fiber-sheet coordinates by using quantitative microstructural measurements (n = 5). RESULTS: A functional border was defined in the midlateral left ventricle; ischemia decreased posterolateral fractional area shortening, and anterolateral fractional area shortening increased. In this demarcation junction, subepicardial end-systolic radial wall thickening decreased (0.16 +/- 0.08 vs 0.11 +/- 0.06) and sheet-normal shear was abolished (0.08 +/- 0.04 vs -0.01 +/- 0.03). Longitudinal shortening decreased in the subepicardium and midwall (-0.05 +/- 0.04 vs +/- -0.01 +/- 0.06), but circumferential-radial shear increased at these depths (0.04 +/- 0.04 vs 0.11 +/- 0.05). Subendocardial fiber stretch occurred during early systole (-0.01 +/- 0.03 vs 0.02 +/- 0.03), and end-systolic fiber-sheet shear increased (0.07 +/- 0.01 vs 0.11 +/- 0.04, all P < .05). CONCLUSIONS: Increased circumferential-radial shear and altered fiber-sheet strains reflect mechanical interactions between ischemic and nonischemic myocardium, which might be important in triggering remodeling processes that evolve into global ischemic cardiomyopathy. PMID- 15821646 TI - Surgical resection for lung cancer with infiltration of the thoracic aorta. AB - OBJECTIVE: The purpose of this study was to evaluate the results of a combined resection of the thoracic aorta and primary lung cancer. METHODS: Sixteen patients underwent thoracic aorta resection along with a left pneumonectomy (n = 6), left upper lobectomy (n = 9), or partial lung resection (n = 1), of whom 10 also received preoperative induction therapy. Cardiopulmonary bypass was used in 10 patients, and a passive shunt between the ascending aorta and the descending aorta was used in 4 patients. RESULTS: Six postoperative major complications occurred in 5 patients, including postoperative bleeding (n = 3), intraoperative bleeding (n = 1), chylothorax (n = 1), and respiratory failure (n = 1). The postoperative morbidity rate was 31%, and the mortality rate was 12.5% (2/16). Furthermore, 4 patients died of systemic tumor relapse, and 1 patient died of intrapleural recurrence. Nine patients were alive after a median follow-up of 54 months (range, 12-199 months). The median survival time of patients with postoperative pathologic N0 disease was 31 months, whereas it was 10 months for those with pathologic N2 or N3 disease. Five-year survivals were 70% for patients with N0 disease and 16.7% for patients with N2 or N3 disease ( P = .0070). CONCLUSIONS: Although pulmonary resection with the involved aorta might cause high surgical morbidity and mortality rates, encouraging long-term survivals were obtained in patients without mediastinal nodal involvement. PMID- 15821647 TI - Long-term graft patency after replacement of the brachiocephalic veins combined with resection of mediastinal tumors. AB - OBJECTIVE: We sought to investigate the correlation between type of vascular reconstruction and long-term graft patency after replacement of brachiocephalic veins combined with resection of mediastinal malignancies. METHODS: Eighteen patients underwent surgical resection of tumors and the superior vena cava with concomitant vascular reconstruction using ringed polytetrafluoroethylene grafts. Graft patency was verified by means of venography or contrast-enhanced computed tomography at time points ranging from 3 to 77 months (median, 33 months) postoperatively. RESULTS: Seven patients underwent sole reconstruction of the right brachiocephalic vein, with occlusion observed in only 1 patient. In 6 patients who underwent reconstruction of the bilateral brachiocephalic veins with 2 separate grafts, the grafts remained patent in 2, whereas 4 patients experienced occlusion of one of the two grafts yet remained asymptomatic. Both patients who underwent reconstruction with a Y graft experienced left brachiocephalic vein graft occlusion. In the 3 patients who underwent reconstruction of a left brachiocephalic vein, the graft became occluded, and superior vena cava syndrome developed in 2 of these patients. CONCLUSION: When replacing the superior vena cava, reconstruction of a left brachiocephalic vein alone results in a significant rate of occlusion and development of superior vena cava syndrome. Thus we advocate sole right brachiocephalic vein reconstruction or bilateral brachiocephalic vein reconstruction in this setting, and separate reconstruction of the veins is preferable to use of a Y graft. PMID- 15821648 TI - Video-assisted wedge resection and local radiotherapy for peripheral lung cancer in high-risk patients: the Cancer and Leukemia Group B (CALGB) 9335, a phase II, multi-institutional cooperative group study. AB - OBJECTIVES: This study examined the feasibility of thoracoscopic wedge resection and radiotherapy for clinical T1 lesions in patients with compromised cardiopulmonary status. METHODS: In this phase II, prospective, multicenter, cooperative group trial, high-risk patients had one or more of the following risk factors: forced expiratory volume in 1 second less than 40%, carbon monoxide diffusing capacity in lung less than 50%, and maximum oxygen consumption less than 45 mm Hg. Patients underwent video-assisted wedge resection followed by local (56 Gy) radiotherapy. The primary end point was the proportion of patients whose disease could be completely resected and who received radiotherapy without treatment complications. RESULTS: Between September 1995 and September 1999, a total of 65 patients were accrued, of which 58 were eligible (52% male, median age 69 years). Pathologic staging resulted in upgrading to T2 or greater in 16 of 58 cases (28%) and in reassessment as benign in 10 of 58 cases (17%). Conversion to thoracotomy was required in 10 cases (17%), including 1 of 10 benign T1-size lesion (10%), 4 of 35 non-small cell lung cancer T1 lesions (13%), and 5 of 14 non-small cell lung cancer T2 lesions (31%). Resection margins were positive in 5 patients: 6% of T1 and 23% of T2. Surgery was aborted in 2 cases (3.5%), and operative mortality was 4%. Overall operative failure rates of video-assisted wedge resection were 20% for benign T1-size lesions, 22% for T1 non-small cell lung cancer, 21% for all T1 lesions, 50% for T2 non-small cell lung cancer, and 29% for all lesions in this study (clinical T1). Prolonged air leaks occurred in 10%, pneumonia in 6%, and respiratory failure in 4%. Thirty-one patients were eligible for radiotherapy; 3 of them refused, and 1 died before treatment. Among the 28 patients who received radiotherapy, severe dyspnea was noted in 3 patients (11%) and moderate pneumonitis in 4 (14%). CONCLUSIONS: Clinical staging in high risk patients is often inaccurate (45% difference from pathologic staging). Intention to treat clinically staged T1 disease by video-assisted wedge resection is associated with a high failure rate. Pathologically staged T1 lesions can be successfully resected in 75% of cases; however, narrow resection margins remain a concern. PMID- 15821649 TI - Large-cell neuroendocrine carcinoma of the lung: a clinicopathologic study of eighteen cases and the efficacy of adjuvant treatment with octreotide. AB - OBJECTIVE: This study was undertaken to evaluate the efficacy of a new adjuvant protocol with octreotide, alone or in combination with radiotherapy, in radically resected large cell neuroendocrine carcinomas of the lung. METHODS: Between 1990 and 2001, a total of 18 consecutive patients affected by large cell neuroendocrine carcinomas of the lung were operated on. Lobectomy and systemic lymphadenectomy were performed in all cases. Postoperative radiotherapy was performed when stage was higher than Ib. Ten patients with positive results of preoperative indium In-111 pentetreotide scintigraphy received octreotide after the operation. RESULTS: Nine patients (50%) had local recurrences or distant metastases (mean recurrence time 14 months); palliative chemotherapy was given, but all patients died. In 10 cases (55.5%) octreotide alone or in combination with radiotherapy was administered as adjuvant treatment; 9 of these patients (90%) are alive and free of disease ( P = .0007), and the other had liver and brain metastases 21 months after surgery. CONCLUSIONS: Our preliminary results seem to demonstrate the efficacy of octreotide as adjuvant therapy in large cell neuroendocrine carcinomas of the lung when results of preoperative indium In-111 pentetreotide scintigraphy were positive. Further study are required to assess the utility of octreotide in patients with negative results of indium In-111 pentetreotide scintigraphy. PMID- 15821650 TI - Border between N1 and N2 stations in lung carcinoma: lessons from lymph node metastatic patterns of lower lobe tumors. AB - OBJECTIVE: Distinction of lymph node stations is one of the most crucial topics still not entirely resolved by many lung cancer surgeons. The nodes around the junction of the hilum and mediastinum are key points at issue. We examined the spread pattern of lymph node metastases, investigated the prognosis according to the level of the involved nodes, and conclusively analyzed the border between N1 and N2 stations. METHODS: We reviewed the records of 604 consecutive patients who underwent complete resection for non-small cell lung carcinoma of the lower lobe. RESULTS: There were 390 patients (64.6%) with N0 disease, 127 (21.0%) with N1, and 87 (14.4%) with N2. Whereas 11.3% of patients with right N2 disease had skip metastases limited to the subcarinal nodes, 32.6% of patients with left N2 disease had skip metastases, of which 64.2% had involvement of N2 station nodes, except the subcarinal ones. The overall 5-year survivals of patients with N0, N1, and N2 disease were 71.0%, 50.8%, and 16.7%, respectively (N0 vs N1 P = .0001, N1 vs N2, P < .0001). Although there were no significant differences in survival according to the side of the tumor among patients with N0 or N1 disease, patients with a left N2 tumor had a worse prognosis than those with a right N2 tumor (P = .0387). The overall 5-year survivals of patients with N0, intralobar N1, hilar N1, lower mediastinal N2, and upper mediastinal N2 disease were 71.0%, 60.1%, 38.8%, 24.8%, and 0%, respectively. Significant differences were observed between intralobar N1 and hilar N1 disease ( P = .0489), hilar N1 and lower mediastinal N2 disease (P = .0158), and lower and upper mediastinal N2 disease (P = .0446). Also, the 5-year survivals of patients with involvement up to station 11, up to station 10, and up to station 7 were 41.4%, 37.9% and 37.7%, respectively (difference not significant). CONCLUSIONS: N1 and N2 diseases appeared as a combination of subgroups: intralobar N1 disease, hilar N1 disease, lower mediastinal N2 disease, and upper mediastinal N2 disease. Interestingly, the survivals of patients with involvement up to interlobar nodes (station 11), main bronchus nodes (station 10), and subcarinal nodes (station 7) were identical. These data constitute the basis for a larger investigation to develop a lymph node map in lung cancer. PMID- 15821651 TI - Off-pump replacement of the pulmonary valve in large right ventricular outflow tracts: a hybrid approach. AB - BACKGROUND: Percutaneous pulmonary valve replacement has recently been introduced and is under investigation in humans. This technique is, however, limited to patients with a right ventricular outflow tract that does not exceed 22 mm in diameter. We report our experience of off-pump pulmonary valve replacement using a hybrid approach in animals with large right ventricular outflow tracts. METHODS: Eight ewes were included in the protocol and were equally divided into 2 groups. A left thoracotomy was first performed, and the main pulmonary artery was banded by using 2 radiopaque rings with a diameter of 18 mm that allowed for further pulmonary valve replacement. We then intended to implant a valved stent either percutaneously (group 1) or through a transventricular approach (group 2). All animals were killed after valve implantation. The operation allowed the pulmonary diameter to be reduced from 30 to 17.6 mm. RESULTS: The right ventricular pressure did not significantly increase after reduction of the pulmonary artery diameter (25 vs 36 mm Hg). Subsequent pulmonary valve replacement through a percutaneous or a transventricular approach was always possible without any requirement for extracorporeal circulation. All devices were successfully delivered inside the pulmonary artery banding and were functioning perfectly at early evaluation. CONCLUSIONS: Implantation of a pulmonary valve is possible in ewes through a hybrid approach when the right ventricular outflow tract exceeds 22 mm in diameter. This involves both surgeons and interventionists and allows for a staged procedure in which the valvulation is performed percutaneously or, for a combined hybrid approach, in which the valve is implanted off pump transventricularly during the same operation. PMID- 15821652 TI - Influence of body core temperature on blood loss and transfusion requirements during off-pump coronary artery bypass grafting: a comparison of 3 warming systems. AB - BACKGROUND: The aim of this prospective randomized trial was to evaluate the efficacy of 3 intraoperative warming systems (Warm-Touch, Thermamed SmartCare OP system, and Allon 2001) on maintenance of normothermia and to investigate their effects on perioperative bleeding and transfusion requirements in patients undergoing off-pump coronary artery bypass grafting. METHODS: With institutional approval/patient informed consent, 90 patients presenting for elective multiple off-pump coronary artery bypass grafting were randomly assigned to 1 of the 3 warming systems. Active warming was started after the induction of anesthesia. Perioperative transfusion was based on international guidelines. Body core temperature was recorded every 30 minutes during operation. Perioperative blood loss, autotransfusion, and allogenic transfusions were recorded. Analysis of variance was performed with post hoc Scheffe tests and chi 2 tests. RESULTS: Normothermia could be sufficiently maintained during operation by the Allon 2001 only. Final body core temperature was 34.7 degrees C +/- 0.9 degrees C (Warm Touch), 35.6 degrees C +/- 0.8 degrees C (Thermamed SmartCare OP), and 36.5 degrees C +/- 0.4 degrees C (Allon 2001; P < .001, Warm-Touch vs Thermamed SmartCare OP, Warm-Touch vs Allon 2001, and Thermamed SmartCare OP vs Allon 2001). Perioperative blood loss was 2683 +/- 1049 mL (Warm-Touch), 2300 +/- 788 mL (Thermamed SmartCare OP), and 1497 +/- 497 mL (Allon 2001; P = .195, Warm Touch vs Thermamed SmartCare OP; P < .001, Warm-Touch vs Allon 2001; P = .001, Thermamed SmartCare OP vs Allon 2001). Transfusion requirements were 1097 +/- 874 mL (Warm-Touch), 986 +/- 744 mL (Thermamed SmartCare OP), and 431 +/- 387 mL (Allon 2001; P = .838, Warm-Touch vs Thermamed SmartCare OP; P = .003, Warm-Touch vs Allon 2001; P = .013, Thermamed SmartCare OP vs Allon 2001). Free of allogenic transfusion were 15 (51.7%; Warm-Touch), 18 (60%; Thermamed SmartCare OP), and 24 (82.8%; Allon 2001) patients ( P = .037). CONCLUSIONS: The goal of normothermia during off-pump coronary artery bypass grafting was best achieved by the Allon system. With this concept, overall blood loss and transfusion requirements were reduced, hence indicating improved quality of perioperative care. PMID- 15821653 TI - Intraoperative identification of esophageal sentinel lymph nodes with near infrared fluorescence imaging. AB - OBJECTIVE: In esophageal cancer, selective removal of involved lymph nodes could improve survival and limit complications from extended lymphadenectomy. Mapping with vital blue dyes or technetium Tc-99m often fails to identify intrathoracic sentinel lymph nodes. Our purpose was to develop an intraoperative method for identifying sentinel lymph nodes of the esophagus with high-sensitivity near infrared fluorescence imaging. METHODS: Six Yorkshire pigs underwent thoracotomy and received submucosal, esophageal injection of quantum dots, a novel near infrared fluorescent lymph tracer designed for retention in sentinel lymph nodes. Six additional pigs underwent thoracotomy and received submucosal esophageal injection of CW800 conjugated to human serum albumin, another novel lymph tracer designed for uptake into distant lymph nodes. Finally, 6 pigs received submucosal injection of the fluorophore-conjugated albumin with an endoscopic needle through an esophagascope. These lymph tracers fluoresce in the near-infrared, permitting visualization of migration to sentinel lymph nodes with a custom intraoperative imaging system. RESULTS: Injection of the near-infrared fluorescent lymph tracers into the esophagus revealed communicating lymph nodes within 5 minutes of injection. In all 6 pigs that received quantum dot injection, only a single sentinel lymph node was identified. Among pigs that received fluorophore conjugated albumin injection, in 5 of 12 a single sentinel lymph node was revealed, but in 7 of 12 two sentinel lymph nodes were identified. There was no dominant pattern in the appearance of the sentinel lymph nodes either cranial or caudal to the injection site. CONCLUSION: Near-infrared fluorescence imaging of sentinel lymph nodes is a novel and reliable intraoperative technique with the power to assist with identification and resection of esophageal sentinel lymph nodes. PMID- 15821654 TI - Activation of neutrophils and monocytes by a leukocyte-depleting filter used throughout cardiopulmonary bypass. AB - OBJECTIVE: Cardiopulmonary bypass elicits systemic inflammation. Depletion of circulating leukocytes might alleviate inflammatory response. We studied the effects of a leukocyte-depleting filter on phagocyte activation during cardiopulmonary bypass. METHODS: Fifty patients undergoing coronary artery bypass grafting were randomly allocated into an arterial line leukocyte filter group (n = 25) with a Pall LeukoGuard 6 leukocyte-depleting filter (LG6; Pall Biomedical, Portsmouth, United Kingdom) and a control group without any filter (n = 25). Blood sampling took place from arterial line at predetermined time points. In the filter group, the sample was taken immediately before the filter; to evaluate activation at the site, an additional sample was taken immediately after the filter. CD11b/CD18 and L-selectin expressions and basal production of hydrogen peroxide were determined with whole-blood flow cytometry, and plasma lactoferrin level was determined with enzyme-linked immunosorbent assay. RESULTS: Neutrophil CD11b expression was higher in the filter group than in the control group (P < .001). Likewise, monocyte CD11b expression, neutrophil hydrogen peroxide production, and lactoferrin plasma levels were all significantly higher, whereas neutrophil and monocyte counts and neutrophil L-selectin expression were all significantly lower in the filter group (all P < .001). At 5 minutes of CPB, CD11b expression increased across the filter on neutrophils (median difference 197 relative fluorescence units, range 45-431 relative fluorescence units, P < .001) and monocytes (median difference 26 relative fluorescence units, range -68 111 relative fluorescence units, P < .001). CONCLUSION: The LG6 arterial line leukocyte filter is ineffective in its principal task of diminishing phagocyte activation during cardiopulmonary bypass. PMID- 15821655 TI - Patient survival characteristics after routine mitral valve repair for ischemic mitral regurgitation. AB - BACKGROUND: Ischemic mitral regurgitation has been associated with diminished survival compared with nonischemic mitral regurgitation. Conversion from mitral valve replacement to valve repair has improved prognosis, but it is unclear whether ischemic mitral regurgitation remains an independent predictor of outcome after mitral valve repair. METHODS: Five hundred thirty-five patients undergoing mitral valve repair (primarily rigid ring annuloplasty) with or without coronary bypass from 1993 through 2002 were reviewed retrospectively (ischemic mitral regurgitation, n = 141; nonischemic mitral regurgitation, n = 394). A Cox proportional hazards model evaluated survival as a function of 9 simultaneous covariates: ischemic versus nonischemic mitral regurgitation, age, sex, number of medical comorbidities, ejection fraction, New York Heart Association class, coronary disease, reoperation, and year of operation. RESULTS: According to univariable analysis, patients with ischemic mitral regurgitation had greater age, higher comorbidity, lower ejection fraction, higher New York Heart Association, and higher reoperation rate (all P < .001) compared with those having nonischemic mitral regurgitation. Univariable 30-day mortality was as follows: 4.3% for patients with ischemic mitral regurgitation versus 1.3% for patients with nonischemic mitral regurgitation (P = .01). Unadjusted 5-year mortality was as follows: 44% +/- 5% for patients with ischemic mitral regurgitation versus 16% +/- 3% for patients with nonischemic mitral regurgitation (P < .001). In the multivariable model, however, only the number of preoperative comorbidities and advanced age were independent predictors of survival (P < .0001), whereas ischemic mitral regurgitation, sex, ejection fraction, New York Heart Association class, coronary disease, reoperation, and year of operation did not achieve significance (all P > .19). After being adjusted for differences in all preoperative risk factors, survival was not statistically different between ischemic mitral regurgitation and nonischemic mitral regurgitation (P = .33). CONCLUSIONS: With routine application of rigid ring annuloplasty, long-term patient survival is more influenced by baseline patient characteristics and comorbidity than by ischemic cause of mitral regurgitation per se. Future risk assessment and decision making should be based on patient condition and should not be biased by ischemic cause of mitral regurgitation. PMID- 15821656 TI - Midterm results of Manouguian double valve replacement: comparison with standard double valve replacement. AB - OBJECTIVE: Surgical strategies for patients who have a small aortic annulus associated with a small mitral annulus remain controversial. The objective of this study was to assess the validity of the Manouguian procedure for double valve replacement with patch enlargement of the annuli. METHODS: We reviewed 57 consecutive patients who underwent double valve replacement for combined aortic and mitral disease between September 1991 and October 2000. Thirty patients underwent the Manouguian procedure with patch enlargement of the aortic and mitral annuli, and the other 27 patients had standard double valve replacement. The Manouguian procedure was selected for a small aortic annulus of less than 21 mm in diameter. The patients were followed for a median of 7.5 years. Results of echocardiography and the dobutamine stress test were compared postoperatively. RESULTS: Double valve replacement with the Manouguian procedure did not increase the early or midterm mortality because the survival (including operative death) at 8 years was 83% in the Manouguian group and 84% in the double valve replacement group (P = .82). The event-free rate at 8 years was 79% in the Manouguian group and 84% in the double valve replacement group (P = .6). The prostheses implanted at the aortic and mitral positions were smaller in the Manouguian group, even after annular enlargement. However, the transprosthetic gradient across the aortic valve evaluated by means of dobutamine stress echocardiography did not differ between the 2 groups, possibly because of a smaller body surface area in the Manouguian group. CONCLUSIONS: The Manouguian patch enlargement procedure is useful during double valve replacement when associated with the problem of small-valve annuli. PMID- 15821657 TI - Mitral valve repair in children with rheumatic heart disease. AB - OBJECTIVE: The purpose of this study was to assess the long-term results of mitral valve repair in children with chronic rheumatic heart disease. METHODS: From January 1988 through December 2003, 278 children (153 male children) underwent mitral valve repair. Mean age was 11.7 +/- 2.9 years (range, 2-15 years). One hundred seventy-three children (62%) were in the New York Heart Association functional class III or IV. Congestive heart failure was present in 24 (8.6%). Reparative procedures included posterior collar annuloplasty (n = 242), commissurotomy (n = 187), cusp-level chordal shortening (n = 94), cusp thinning (n = 71), cleft suture (n = 65), and cusp excision or plication (n = 10). Associated procedures included atrial septal defect closure (n = 22), aortic valve repair/replacement (n = 13), and tricuspid valve repair (n = 3). RESULTS: Early mortality was 2.2% (6 patients). Preoperative left ventricular dysfunction was associated with greater mortality. Median follow-up was 56.5 months (mean, 58.9. +/- 32.3 months; range, 5 to 180 months). One hundred seventy-seven survivors (65%) had no or trivial mitral regurgitation. Sixteen patients (6%) required reoperation for valve dysfunction. There were 7 late deaths (2.6%). Actuarial, reoperation-free, and event-free survivals at a median follow-up of 56.5 months were 95.2% +/- 1.5%, 91.6% +/- 2.2%, and 55.9% +/- 3.5%, respectively; at 15 years, they were 95.2% +/- 1.5%, 85.9% +/- 5.9%, and 46.7% +/ 4.7%, respectively. CONCLUSION: Mitral valve repair in children with chronic rheumatic heart disease is feasible and provides acceptable long-term results. PMID- 15821658 TI - Traumatic rupture of the thoracic aorta: ten years of delayed management. AB - OBJECTIVE: Traumatic rupture of the thoracic aorta is a highly fatal condition in which patient outcome is strongly conditioned by other associated injuries. Delayed aortic treatment has been proposed to improve results. METHODS: The charts of 69 patients with traumatic rupture of the thoracic aorta observed between 1980 and 2003 were reviewed. Patients were grouped according the timing of repair: group I, immediate repair (21 patients); and group II, delayed repair (48 patients). In group II, 45 patients were treated surgically or by endovascular procedure. RESULTS: In-hospital mortalities were 4 of 21 patients (19%) in group I and 2 of 48 patients (4.2%) in group II. There were 3 cases of paraplegia in group I and none in group II. CONCLUSION: Improvement of patient outcome with traumatic rupture of the thoracic aorta can be achieved by delaying surgical repair until after management of major associated injuries if there are no signs of impending rupture. Endovascular treatment is feasible and safe and may represent a valid alternative to open surgery in selected cases. PMID- 15821659 TI - Determining the best procedure for radial artery harvest: prospective randomized trial for early postharvest complications. AB - OBJECTIVE: Although useful procedures for radial artery harvest have been reported, forearm circulation and collateral perfusion after radial artery harvesting remain unknown. To assess an optimal radial artery harvest technique for forearm circulation, we designed a prospective randomized trial. METHODS: Ninety patients were divided into 3 groups of 30 patients. Electrocautery, an ultrasonic scalpel, or sharp scissors and hemoclips were used to harvest radial arteries in groups 1, 2, and 3, respectively. The incidences of harvest site pain, numbness, swelling, discomfort, hematoma, and infection were compared. With forearm thermography, recovery times from cooled down 5 degrees back to rest temperature were compared between groups. All patients had postoperative forearm angiography at 1 and 12 months. RESULTS: Although there were no differences in the incidences of pain, swelling, and discomfort, the incidence of numbness was significantly lower in group 3 (P = .003). The temperature recovery time was significantly shorter in group 3 (P = .0009). On postoperative angiography at 1 month, the incidence of the development of interosseous arteries was significantly higher in group 3 (86.7%) than in groups 1 (23.3%) and 2 (36.7%). The 12-month study, however, showed that there was no difference among groups (73.3%, 80.0%, and 93.3% in groups 1, 2, and 3, respectively). CONCLUSIONS: These results suggest that sharp dissection with scissors and clips may be better for early postharvest forearm circulation and can decrease the incidence of hand numbness. However, there were no differences among the 3 methods with respect to forearm circulation 12 months after radial artery harvest. PMID- 15821660 TI - Preoperative moderate to severe diastolic dysfunction: a novel Doppler echocardiographic long-term prognostic factor in patients with severe aortic stenosis. AB - OBJECTIVE: We studied long-term outcomes in severe aortic stenosis and the importance of prosthesis type (mechanical vs biologic) and size, preoperative left ventricular ejection fraction, diastolic function, and left ventricular mass. METHODS: Patients undergoing valve replacement from 1991 to 1993 (n = 399, 45% women) were included. The diastolic function was evaluated by integrating mitral and pulmonary venous flow data with Doppler echocardiography. The patients were classified as having either normal diastolic function to mild diastolic dysfunction or moderate to severe diastolic dysfunction. Left ventricular ejection fraction and the diastolic function category were incorporated together with age, sex, and time since operation into a Poisson regression model with death as the end point. Prosthesis type and size and left ventricular mass were also investigated. RESULTS: The age (mean +/- SD) was 71 +/- 9 years, and the overall survival after 12 years was 50%. Although markedly reduced during the initial 6-month period, mortality risk subsequently increased more than could be explained by age (hazard ratio of 1-year difference = 1.12, P = .0005). The moderate to severe diastolic dysfunction pattern independently predicted late mortality (hazard ratio = 1.72, P = .0038), whereas left ventricular ejection fraction did not (hazard ratio = 0.99, P = .18). The prognostic importance of moderate to severe diastolic dysfunction did not diminish with time; on the contrary, it tended to increase. Mortality after 12 years was not predicted by left ventricular mass (P = .66), prosthesis type (P = .57), or prosthesis size (P = .58). CONCLUSION: This study reveals that moderate to severe diastolic dysfunction in patients with aortic stenosis is an independent predictor of late mortality after valve replacement and that its importance does not decrease with time. Our findings may suggest that moderate to severe diastolic dysfunction implies nonreversible myocardial changes that negatively affect survival. PMID- 15821661 TI - Optimizing revascularization strategies in patients with multivessel coronary disease: impact of intracoronary pressure measurements. AB - OBJECTIVES: In patients with multivessel coronary disease, the functional significance of each lesion is often unclear, and preinterventional stress tests may be inconclusive. In this setting, intracoronary pressure measurements may be helpful to define the optimal revascularization strategy. METHODS: Twenty-five consecutive patients (aged 64 +/- 11 years) with multivessel disease, inconclusive stress tests or not performed stress tests, and an angiographically intermediate coronary artery stenosis in at least 1 major vessel underwent intracoronary pressure measurements. Myocardial fractional flow reserve was measured for the intermediate lesions under the condition of maximum hyperemia induced by intravenous adenosine (140 microg x kg(-1) x min(-1). Revascularization strategies based on angiographic information alone were compared with treatment strategies based on fractional flow reserve results. RESULTS: The original recommendation of the revascularization procedure of choice (bypass operation or angioplasty) was changed in 9 patients (36%) on the basis of the results of fractional flow reserve measurements. In 6 more patients, pressure measurements led to a change in the recommended number of anastomoses to be aimed for during the operation. Within diffusely diseased vessels, fractional flow reserve provided an exact segmental resolution of pathologic vessel resistance for optimal graft placement. Significant left main disease was confirmed in 3 of 6 patients and was detected in 3 angiographically unsuspected cases. CONCLUSIONS: In patients with multivessel disease, coronary pressure-derived fractional flow reserve is a valuable tool to guide clinical decision making and support cardiologists and cardiovascular surgeons in the composition of optimal revascularization strategies. PMID- 15821662 TI - Quantitative analysis of survival of transplanted smooth muscle cells with real time polymerase chain reaction. AB - BACKGROUND: Cell transplantation improves heart function after myocardial infarction. This study investigated the survival of implanted cells in normal and infarcted myocardium. METHODS: Male rat aortic smooth muscle cells were cultured. For the in vitro study, male smooth muscle cells mixed with female smooth muscle cells or male smooth muscle cells injected into a piece of female rat myocardium were used to evaluate the accuracy of quantitative real-time polymerase chain reaction to measure Y chromosomes. For the in vivo study, 2 million live or dead male smooth muscle cells were injected into normal or infarcted female myocardium. At 1 hour and 1 and 4 weeks after transplantation, hearts, lungs, and kidneys were harvested for measurement of Y chromosomes. RESULTS: In vitro, the accuracy of polymerase chain reaction measurement was excellent in cultured cells (r2 = 0.996) and the myocardium (r2 = 0.786). In vivo, 1 hour after 2 x 10(6) cell implantation, live cell numbers decreased to 1.0 +/- 0.2 x 10 6 and 1.1 +/- 0.3 x 10(6) , and dead cell numbers decreased to 0.9 +/- 0.2 x 10(6) and 0.8 +/- 0.2 x 10(6) in the normal and infarcted myocardium, respectively (P < .01 for all groups). Lungs and kidneys contained 8.5% and 1.5% of the implanted cells, but no cells were detected at 1 week. At 1 week, no dead smooth muscle cells were detected in the normal or infarcted myocardium. The numbers of live cells at 1 and 4 weeks were 0.48 +/- 0.06 x 10(6) and 0.27 +/- 0.07 x 10(6) in normal myocardium and 0.29 +/- 0.08 x 10(6) and 0.18 +/- 0.05 x 10(6) in infarcted myocardium. CONCLUSIONS: One hour after implantation, only 50% of smooth muscle cells remained in the implanted area. Some implanted cells deposited in other tissue. Implanted cell survival progressively decreased during the 4-week study. PMID- 15821663 TI - High central venous pressure is associated with prolonged mechanical ventilation and increased mortality after lung transplantation. AB - BACKGROUND: Poor oxygenation might occur in transplanted lungs as a result of reperfusion injury and lack of lymphatic drainage. Low central venous and pulmonary capillary wedge pressures are advocated to reduce pulmonary edema and maximize oxygenation but might adversely affect cardiac index, circulation, and renal function. METHODS: Histories, intensive care unit charts, and donor data on 118 lung transplantations performed between 1999 and 2002 were retrospectively assessed. Multiple logistic regression analysis was performed on donor, recipient, operative, and intensive care unit parameters to determine the relationship of filling pressure (central venous and pulmonary capillary wedge pressures) to prolonged mechanical ventilation and outcome. The mean central venous pressure was used to divide patients into high and low central venous pressure groups, which were then compared to determine differences in outcome and complication rates. RESULTS: A high central venous pressure was found to be associated with prolonged mechanical ventilation (odds ratio, 1.57; 95% confidence interval, 1.13-2.20; P = .008). After removing the effect of poor myocardial function by excluding patients with low cardiac index (< 2.2 L x min 1 x m(-2) ) and high inotrope requirement (> 10 microg/min), central venous pressure remained associated with prolonged mechanical ventilation (odds ratio, 2.31; 95% confidence interval, 1.31-4.07; P = .004). Duration of ventilation (P < .001), intensive care unit mortality (P = .02), hospital mortality (P = .09), and 2-month mortality (P = .02) were higher in patients with central venous pressures of greater than 7 mm Hg. There was no evidence of complications caused by hypovolemia in the low (< or = 7 mm Hg) central venous pressure group, who had lower inotrope requirements (P = .02) and lower creatinine levels (P = .013). Conclusions A high central venous pressure was associated with adverse outcomes after lung transplantation. PMID- 15821664 TI - Lung transplantation with lungs from donors fifty years of age and older. AB - BACKGROUND: A shortage of donors has led to the progressive expansion of criteria for donor selection in lung transplantation. The outcome of recipients of lungs from donors aged 50 years or older is analyzed systematically. METHODS: From March 1998 to June 2003, 49 recipients received lungs from donors aged 50 years or older (range 50-64 years, mean 54 +/- 3 years). This group of recipients was compared with 244 patients receiving lungs from donors aged less than 50 years (range 7-49 years, mean 32 +/- 11 years). This study was undertaken on all 293 patients at our institution who received Perfadex-preserved lungs (Vitrolife, Goteborg, Sweden). RESULTS: Recipient age, sex, and indications for transplant did not differ significantly between groups. Also, the percentage of the different types of transplants (bilateral or single lung transplantation) performed was equal in both cohorts. Donor Pa(O2) /F(IO2) ratios before lung retrieval (415 +/- 91 vs 439 +/- 113, respectively) and length of ischemic time (347 +/- 67 minutes vs 351 +/- 84 minutes, respectively) did not differ significantly between the older and younger donor groups. The following posttransplant parameters were also not statistically different: first Pa(O2)/F(IO2) at intensive care unit arrival (274 +/- 125 in the older donor group vs 253 +/- 119 in the younger donor group, respectively), mechanical ventilation time (328 +/- 427 hours vs 269 +/- 425 hours, respectively), and length of stay in the intensive care unit (16 +/- 18 days vs 14 +/- 18 days, respectively). Recipient survival in the older and younger donor groups at 30 days, 3, 6, 12, 24, and 60 months was 77% +/- 6%, 75% +/- 6%, 73% +/- 7%, 73% +/- 7%, 68% +/- 5%, and 68% +/- 4% versus 86% +/- 2%, 83% +/- 3%, 80% +/- 3%, 78% +/- 3%, 71% +/- 4%, and 66% +/- 4%, respectively. CONCLUSIONS: Lung grafts from elderly donors have been considered as marginal organs for transplantation. However, this study indicates that transplantation of lungs from carefully selected donors aged 50 years or more may lead to similar short- and long-term outcomes compared with lungs from younger donors. The use of lungs from elderly donors may help to increase the number of donor organs in lung transplantation. PMID- 15821665 TI - Recipient intramuscular cotransfection of transforming growth factor beta1 and interleukin 10 ameliorates acute lung graft rejection. AB - OBJECTIVE: Multiple gene transfer might permit modulation of concurrent biochemical pathways involved in acute lung graft rejection. We investigated whether gene cotransfection into the recipient reduces acute lung graft rejection. METHODS: Brown Norway rats were used as donors, and F344 rats were used as recipients. Recipient animals were injected with saline (groups I/VI) or 1 x 10(10) pfu of adenovirus encoding beta-galactosidase (groups II/VII), transforming growth factor beta1 (groups III/VIII), interleukin 10 (groups IV/IX), or both transforming growth factor beta1 and interleukin 10 (groups V/X) into both leg muscles 2 days before transplantation (groups I-V) or at the time of harvest (groups VI-X). The Kruskal-Wallis test for rejection score and 1-way analysis of variance were used to compare groups. RESULTS: Oxygenation was significantly improved in the cotransfected groups treated 2 days before transplantation and at the time of harvest. Rejection scores were also reduced in the cotransfected groups. In group V cotransfection suppressed endogenous interleukin 2 but not interferon gamma and tumor necrosis factor alpha. CONCLUSION: Recipient intramuscular cotransfection of transforming growth factor beta1 and interleukin 10 suppressed interleukin 2 expression and provided a synergistic effect that reduced acute lung graft rejection. This approach might be applied to the clinical setting because transplant recipients could be treated at the time of implantation. PMID- 15821666 TI - Total arterial revascularization in a child with familial homozygous hypercholesterolemia. PMID- 15821667 TI - Introduction of off-pump coronary artery bypass into aortic arch aneurysm repair: a new solution for the surgical treatment of multiorgan arteriosclerosis. PMID- 15821668 TI - A cautionary note regarding long-term sequelae of biologic glue. PMID- 15821669 TI - Paradoxical hemopericardium in type B acute aortic dissection. PMID- 15821670 TI - Reoperative innominate arterial, ascending aortic, and root replacement for extensive fungal endocarditis. PMID- 15821671 TI - Tracheal compression caused by an innominate artery aneurysm after thoracic aortic aneurysm repair in a patient with Marfan disease. PMID- 15821672 TI - Mycotic innominate artery pseudoaneurysm presenting as an embolic stroke. PMID- 15821673 TI - Uniportal video-assisted thoracoscopic surgery wedge lung biopsy in the diagnosis of interstitial lung diseases. PMID- 15821674 TI - Early primary Kawashima operation combined with direct hepatic vein-to-azygos vein connection: a new logical approach. PMID- 15821675 TI - Remodeling operation for unruptured aneurysms of three sinuses of Valsalva. PMID- 15821676 TI - Aneurysm of a right-sided aortic arch and right descending aorta: three dimensional volume rendering of multislice computed tomographic aortography facilitates surgical planning and management. PMID- 15821678 TI - Reliability of diagnostic tests in cases of delayed tooth eruption. PMID- 15821679 TI - More research needed to understand how orthodontists communicate with cells. PMID- 15821680 TI - Superimposed tracings show questionable results. PMID- 15821681 TI - Superimposed tracings show questionable results. PMID- 15821684 TI - Intraoral aging of orthodontic materials: the picture we miss and its clinical relevance. AB - The purpose of this editorial is to systematically analyze the variety and potency of various aging variables affecting the morphology, structure, and mechanical properties of polymeric and metallic orthodontic materials. The effects of aging on the longevity of the bond strength and mechanotherapy were analyzed: aging-induced plasticization of resin adhesives might lead to bond failure at forces of magnitude lower than those sustained at the initiation of treatment. Standard in vitro methodologies cannot show this effect, and thus laboratory bond strength protocols require modification to become clinically meaningful. Also, the force transferred from an activated archwire to a preadjusted bracket slot, as well as friction during free sliding, seems to be affected by the intraorally induced alteration of materials. Although the effect of intraoral environmental conditions on the superelastic properties of nickel titanium (Ni-Ti) archwires and coil springs requires further research to establish the true spectrum of effects, it has been suggested that intraoral temperature variations might transiently affect their properties and that the fracture resistance of used Ni-Ti wires is reduced. Clinical implications are discussed for (1) in vivo-aged elastomeric ligatures and chains, which can be postulated to express much higher creep than their in vitro-aged counterparts; (2) the largely unknown effect of aging on the spring component of self-ligated brackets and the associated effect on ligation force; and (3) the intraorally induced alterations in the structural conformation of Invisalign appliances (Align Technology, Santa Clara, Calif). The objective of future research efforts in the field of orthodontic materials should include the development of clinically relevant methodologies. A clear definition of limitations of laboratory experimental configurations might be instrumental in confining the clinical impact of research findings to their actual extent. PMID- 15821685 TI - Light-cured or chemically cured orthodontic adhesive resins? A selection based on the degree of cure, monomer leaching, and cytotoxicity. AB - PURPOSE: The purpose of this study was to estimate the degree of cure and monomer leaching of a light-cured and a chemically cured (no-mix) adhesive and to assess their biologic properties. MATERIAL: The degree of cure of adhesive specimens prepared with a procedure identical to the clinical bonding process was assessed by infrared spectroscopy. The adhesives were then immersed in normal saline solution for 2 months, and the residual monomer leached from the adhesives was quantitatively and qualitatively analyzed by liquid chromatography. The effect of the immersion media on human gingival fibroblasts' viability and proliferation was also evaluated with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and deoxyribonucleic acid (DNA) synthesis assays, respectively. The results were analyzed by analysis of variance and Tukey test (alpha = .05). RESULTS: No difference was found between the 2 adhesives with respect to their degree of cure and the amount of triethylene glycol dimethacrylate released; no diglycidyl dimethacrylate monomer was detected in the eluent. However, significant qualitative changes in the composition of the substances eluted from the 2 adhesives were observed. CONCLUSIONS: Whereas no cytotoxic effect was shown for either immersion media, a moderate reduction in the DNA synthesis was obtained by both adhesives, implying a minor cytostatic effect. Further research is required to assess the long-term biologic properties of adhesives, including potential estrogenic action. PMID- 15821686 TI - Degradation of plastic polyoxymethylene brackets and the subsequent release of toxic formaldehyde. AB - PURPOSE: Heat, acids, alkalis, oxygen, abrasion, enzymes, and radiation are all viable mechanisms for the chemical breakdown of polyoxymethylene (POM), a plastic material used in some esthetic orthodontic brackets. The aim of this study was to establish the thermal characteristics of POM brackets and the chemical by products in the as-received bracket, during thermal analyses of the bracket, and after abrasion of the slot and base of the bracket. MATERIAL: Plastic brackets and control rods made of POM were evaluated ex vivo by thermal and chemical analyses. RESULTS: POM brackets produce toxic formaldehyde gas whether heated or mechanically abraded. Patients who wear these brackets are being exposed to, at the very least, a potential irritant. Thermal analyses showed that the melting temperature of a POM bracket was approximately 178 degrees C, comparable to that for a commercial rod of POM. Both POM products started to degrade at approximately 250 degrees C, and, by 420 degrees C, both products had completely decomposed into their fundamental molecular structure, formaldehyde. A colorimetric assay with a fuchsin-aldehyde reagent (Schiff's reagent) showed that aldehydes were present in the effluent from thermal heating, from mechanical abrasion of the bracket's slot or base, and even from the as-received bracket. The only difference between the 6 to 8-week assays and the 12 to 14-week assays was the intensification of the color, which occurred because of an increase in aldehyde formation and the concomitant increase in double-bond formation. These observations are consistent with a report in the tribologic (ie, friction and wear) literature, which documented the degradation of POM when it rubs against steel. CONCLUSIONS: Because formaldehyde's inherent uses as a disinfectant base and embalming fluid preclude its beneficial presence in the human body, further use of POM for orthodontic brackets, crowns for children, and other prosthetic appliances is contraindicated because even radiography will promote its degradation. PMID- 15821687 TI - Effect of 0.4% stannous fluoride gel on Streptococci mutans in relation to elastomeric rings and steel ligatures in orthodontic patients. AB - BACKGROUND: Patients with fixed orthodontic appliances often experience an absolute increase in the number of Streptococci mutans colony-forming units (cfu). The aim of this investigation was to study the development of biofilm and S. mutans cfu in connection with stainless steel ligatures and elastomeric rings in orthodontic patients treated with and without 0.4% stannous fluoride gel (SFG). MATERIAL: Forty-seven patients were divided into 2 groups: those treated with 0.4% SFG for 4 minutes (experimental) and those without 0.4% SFG (control). In each patient, elastomeric rings were used for ligation on 1 side of the dental arch midline, and stainless steel ligatures were used on the opposite side. Saliva samples were collected before and after appliance placement. At 15 and 30 days after appliance placement, biofilm samples from the stainless steel ligatures and the elastomeric rings were collected and subjected to microbiologic procedures and scanning electron microscopy (SEM) analysis. RESULTS: The numbers of S. mutans cfu in the saliva and biofilm were not statistically different between the teeth fitted with elastomeric rings and stainless steel ligatures, or between the experimental and control groups. SEM analysis showed biofilm formation on both ligature ties. CONCLUSIONS: Topical application of 0.4% SFG in orthodontic patients with elastomeric rings or stainless steel ligatures does not cause a significant decrease in S. mutans cfu in the saliva and biofilm. PMID- 15821688 TI - Clinical assessment of orthodontic outcomes with the peer assessment rating, discrepancy index, objective grading system, and comprehensive clinical assessment. AB - PURPOSE: The purpose of this study was to quantitatively assess orthodontic treatment outcomes in postgraduate orthodontic clinics at Okayama University (OU) and Indiana University (IU). MATERIAL: Using the peer assessment rating (PAR) index, the discrepancy index (DI), the American Board of Orthodontist's objective grading system (OGS), and the comprehensive clinical assessment (CCA), we evaluated pretreatment and posttreatment records of 72 patients from OU and 54 patients from IU. RESULTS: The average pretreatment PAR score with United Kingdom weighting was 32 for OU subjects and 28 for IU subjects. Differences in maxillary and mandibular buccal alignment between schools were statistically significant ( P < .01). The posttreatment PAR scores were 7 for OU and 4 for IU. The difference in overjet between schools was statistically significant ( P < .05). The mean DI scores were 19 for OU and 17 for IU. OU patients scored significantly more DI points for crowding and mandibular plane angle compared with IU patients ( P < .05). On the other hand, they lost significantly fewer DI points for overbite and occlusion compared with IU patients ( P < .05). The mean OGS scores were 34 for OU and 33 for IU. Buccolingual inclination and overjet scores were significantly higher in OU patients compared with IU ( P < .05). The mean CCA score was approximately 4 points for both OU and IU. CONCLUSIONS: These data suggest that these indexes are useful for comparing treatment outcomes between clinics. They were able to identify specific problems in treating Asian patients. PMID- 15821689 TI - Long-term posttreatment changes measured by the American Board of Orthodontics objective grading system. AB - BACKGROUND: The American Board of Orthodontics (ABO) developed the objective grading system (OGS) to evaluate cases submitted as part of the phase III examination for board certification. This study examines the long-term changes in 6 of the 8 criteria measured by the board, with an emphasis on the relationship between posttreatment results and subsequent improvement or worsening. MATERIAL: One hundred subjects were randomly chosen from the postretention archives of the Department of Orthodontics at the University of Washington. Pretreatment peer assessment rating (PAR) scores and posttreatment and postretention OGS scores were measured on study casts. The ABO criteria studied were alignment, marginal ridges, buccolingual inclinations, occlusal contacts, occlusal relationships, and overjet. Interproximal contacts and root angulations were not scored. RESULTS: The mean overall OGS score at posttreatment was 21.5 points. At postretention, this score had improved significantly, by approximately 4 points. When comparing posttreatment scores and long-term changes, similar patterns were seen for all criteria, except alignment; increased posttreatment scores were associated with increased long-term improvement. Well-treated cases tended to deteriorate, and poorly finished cases tended to improve, illustrating a regression to the mean. Alignment was the only criterion associated with a mean long-term worsening, as well as a less predictable pattern of change. CONCLUSIONS: The results of this study indicate that settling occurs after orthodontic treatment, and that the attainment of perfect occlusal results does not ensure stability. PMID- 15821690 TI - Long-term comparison of treatment outcome and stability of Class II patients treated with functional appliances versus bilateral sagittal split ramus osteotomy. AB - PURPOSE: The objective of this study was to compare the treatment outcomes and stability of patients with Class II malocclusion treated with either functional appliances or surgical mandibular advancement. MATERIAL: The early-treatment group consisted of 30 patients (15 girls, 15 boys), with a mean age of 10 years 4 months (range, 7 years 5 months to 12 years 5 months), who received either Frankel II (15 patients) or Herbst appliances (15 patients). The surgical group consisted of 30 patients (23 female, 7 male), with a mean age of 27 years 2 months (range, 13 years 0 months to 53 years 10 months). They were treated with bilateral sagittal split ramus osteotomies with rigid fixation. Lateral cephalograms were taken for the early-treatment group at T1 (initial records), T2 (completion of functional appliance treatment), and Tf (completion of comprehensive treatment). In the surgical group, lateral cephalograms were taken at T1 (initial records), T2 (presurgery), T3 (postsurgery), and Tf (completion of comprehensive treatment). The average times from the completion of functional appliance treatment or surgery to the final cephalograms were 35.8 months and 34.9 months, respectively. A mixed-design analysis of variance was used to compare changes within and between groups. RESULTS: In the functional appliance group, the mandible continued to grow in a favorable direction even after discontinuation of the functional appliance. Both groups had stable results over time. Both groups finished treatment with the same cephalometric measurements. Significant skeletal and soft tissue changes were noted in the treatment groups due to either functional or surgical advancement of the mandible. More vertical relapse was noted in the surgical group than in the functional group. CONCLUSIONS: This study suggests that early correction of Class II dentoskeletal malocclusions with functional appliances yields favorable results without the possible deleterious effects of surgery. PMID- 15821691 TI - Prospective clinical trial comparing the effects of conventional Twin-block and mini-block appliances: Part 1. Hard tissue changes. AB - PURPOSE: The aim of this prospective clinical trial was to compare the dentoskeletal effects of a conventional and a modified Twin-block (TB) appliance. The conventional TB appliance was constructed with a large, single-step advancement. The modified appliance, termed the mini-block (MB), was incrementally advanced, incorporated a maxillary incisor torquing spring, and had a reduced bite-block height. MATERIAL: Seventy patients were placed into age- and sex-matched pairs. Patients in each pair were allocated to opposing appliance groups. Active treatment lasted 9 months, irrespective of the final overjet attained, and final cephalometric records were taken at 12 months (+/-1 month). Both groups showed pretreatment equivalence for age, sex, overjet, and cephalometric variables. RESULTS: The TB group experienced a significantly greater reduction in overjet (median, -8 mm; P = .02) compared with the MB group (median, -4 mm). This improved overjet reduction was associated with significantly greater forward movement of pogonion (median change, TB: 3.3 mm; MB: 2.1 mm; P = .02) and greater retroclination of the maxillary incisors (median change, TB: -5 degrees ; MB: -1.9 degrees ; P = .04). No significant intergroup difference was found for changes in total anterior facial height (median change, TB: 4.4 mm; MB: 4.3 mm) and mandibular incisor proclination (median change, TB: 1.3 degrees ; MB, 2.4 degrees ). CONCLUSIONS: Progressive mandibular advancement was not associated with greater mandibular growth compared with single-step advancement. The maxillary incisor torquing spring seems to be effective at reducing maxillary incisor retroclination. Reduced bite activation in the MB group did not result in less mandibular incisor proclination. There was considerable individual variation in appliance effects within both groups. PMID- 15821692 TI - Prospective clinical trial comparing the effects of conventional Twin-block and mini-block appliances: Part 2. Soft tissue changes. AB - PURPOSE: The aims of this prospective clinical trial were to compare the soft tissue effects of Twin-block (TB) and mini-block (MB) functional appliances by using lateral cephalograms and optical surface laser scans and to evaluate the extent of posttreatment relapse with both appliances. MATERIAL: Seventy Class II Division 1 patients meeting set inclusion criteria were matched by age and sex. The matched pairs were randomly allocated to treatment with either the TB or the MB appliance. The appliances were worn full time for 9 months (phase I), followed by a 3-month observation phase of no appliance wear (phase II). Lateral cephalograms were taken at the start of treatment and at 12 months, and laser scans were taken at 0, 3, 9, and 12 months. RESULTS: Radiographically, there was a greater advancement of the soft tissue pogonion in the TB group (median, TB: 4.0 mm; MB: 1.8 mm; P = .004), whereas the soft tissue anterior face height increased similarly in both groups (median, TB: 4.4 mm; MB: 4.3 mm). Optical surface scans confirmed the cephalometric changes. There were a greater forward movement of soft tissue pogonion in the TB group during active treatment (median change, TB: 3.2 mm; MB: 3.9 mm) and similar increases in the soft tissue total anterior face height (median change, TB: 3.2 mm; MB: 3.9 mm). There were negative changes of soft tissue pogonion (median, TB: -1.0 mm; MB: -0.9 mm) and vertical face height (median change, TB: -0.7; MB: -0.6) during phase II. CONCLUSIONS: The TB appliance produced the greater overall change in the soft tissue profile. However, there were clinically significant relapse changes in the immediate postfunctional phase. PMID- 15821693 TI - Skeletal and dental response to rapid maxillary expansion with 2- versus 4-band appliances. AB - PURPOSE: Banding a rapid palatal expansion (RPE) appliance to the 2 first molars has been proposed as an equivalent to the conventional 4-band RPE appliance. However, the dentoskeletal response to this treatment has not been evaluated. MATERIAL: Twenty-eight subjects (aged 8-20 years) who required maxillary expansion were matched by age and randomly assigned to either a 2-band or a 4 band RPE group. Skeletal and dental responses were measured from standardized anteroposterior cephalometric and occlusal radiographs and dental casts before treatment (T1), at the end of expansion (T2), and at 1 year postexpansion (T3). RESULTS: At T2, suture expansion was 2.5 times greater and arch perimeter was 6 fold larger in the 4-band than in the 2-band group. Both appliances displayed the typical "V" expansion of the suture and "reverse V" expansion of the dental arch. At T3, remineralization reduced the suture opening by 75%, but 95% to 99% of the dental reaction was maintained. The 2-band group demonstrated a significant inverse age-dependent correlation ( r = -0.795), with treatment failure beyond 12 years of age, whereas the 4-band group was age-independent. CONCLUSIONS: The greater the skeletal resistance, the smaller the sutural response but the greater the dental response to RPE therapy. Four-band RPE is indicated when severe anterior crowding is accompanied by a tapered arch form, and 2-band RPE is recommended in the mixed dentition when mild crowding occurs with posterior constriction. PMID- 15821695 TI - Treatment of a severe Class III open bite. PMID- 15821694 TI - Maxillary distraction osteogenesis to treat maxillary hypoplasia: comparison of an internal and an external system. AB - BACKGROUND: Distraction osteogenesis (DO) has become a mainstream surgical technique for patients with jaw deformities. In cases of maxillary hypoplasia, DO with a rigid external distraction (RED) system has been used for maxillary advancement; however, DO with internal devices is currently popular. MATERIALS: This article describes DO with an internal device and a RED system in 2 patients with maxillary hypoplasia with oligodontia. The first patient, a young girl, had a concave profile due to maxillary hypoplasia and 9 congenitally missing permanent teeth. At age 11 years 11 months, she received DO with an internal device. The second patient, a boy aged 11 years 7 months, was treated with DO with a RED system. RESULTS: In the girl, the maxilla was advanced 5.0 mm without any dentoalveolar compensation. In the boy, the maxilla was advanced 7.0 mm, but undesirable mesial movement of posterior teeth was observed. CONCLUSIONS: DO with internal devices is simpler and more useful than the RED system for maxillary hypoplasia with oligodontia. PMID- 15821696 TI - Orthodontics in 3 millennia. Chapter 2: entering the modern era. AB - In the early 1900s, groundbreaking scientists, working without benefit of graduate training and having little precedent, built the orthodontic edifice brick by brick. Kingsley pioneered cleft-palate treatment. Case showed us the importance of facial esthetics. Dewey and Ketcham made the ABO the first certifying board in dentistry. But it was Angle who gave us our first school, journal, society, and practical classification of malocclusion. PMID- 15821697 TI - New, fast, and accurate procedure to calibrate a 2-dimensional digital measurement method. AB - Traditionally, dental casts have been measured by hand, with conventional calipers. Today, several digital methods are available for automatically measuring tooth size, but their accuracy depends on proper calibration. The purpose of this article is to introduce a fast and accurate procedure to calibrate a previously introduced 2-dimensional computerized system of measuring mesiodistal tooth size and calculating overall and anterior Bolton ratios. In this calibration method, the cast is digitized while sitting on a sheet of graph paper. The magnification of the paper in 2 axes can be easily calculated and hence the magnification of the dental cast digital image. PMID- 15821698 TI - Litigation, legislation, and ethics. Right to refuse treatment. PMID- 15821699 TI - Hereditary colon cancer--part I. PMID- 15821700 TI - [Combination of prehospital systemic thrombolytic therapy with endovascular procedures in the treatment of patients with acute myocardial infarction]. AB - Since July 2002 we have been conducting a study of efficacy of prehospital thrombolytic therapy combined with subsequent endovascular procedures in the treatment of patients with acute myocardial infarction. Fifty nine patients received prehospital fibrinolysis with tissue-type plasminogen activator (TPA, n=28) or streptokinase (n=31) within 6 hours after onset of symptoms. TPA infusion compared with that of streptokinase was associated with smaller ischemic myocardial damage and lower frequency of side effects (3.6 and 38.7%, respectively). Angioplasty or stenting of infarct related arteries were carried out in 47 of these patients. The group of patients subjected to endovascular interventions was characterized by a low rate of in-hospital cardiac events and zero mortality. PMID- 15821701 TI - [Assessment of coronary reserve in patients with hypertensive disease and without hypertrophy left ventricular]. AB - AIM: To study relationship between coronary reserve and left ventricular geometry. METHOD AND MATERIAL: Transesophageal cardiac pacing was carried out in 53 patients with hypertensive disease. Thirty five patients (66%) had left ventricular hypertrophy which was eccentric in 16 and concentric in 19. RESULTS: Myocardial ischemia was induced during pacing in 79.2% of patients; it was painful in 45.2 and painless -- in 54.8% of patients. Test with esophageal pacing was positive in 91.4 and 55.6% of patients with and without left ventricular hypertrophy, respectively. In patients with concentric hypertrophy frequency of positive tests was higher and level of coronary reserve lower than in patients with eccentric left ventricular hypertrophy. There was negative correlation between pacing rate at myocardial ischemia induction and left ventricular myocardial mass index. Painless ischemia was more frequent among patients with left ventricular hypertrophy. Twelve of 42 patients (28.3%) with positive result of pacing had no clinical signs of ischemic heart disease. CONCLUSION: Left ventricular hypertrophy limits coronary reserve, increases prevalence of painless myocardial ischemia. Transesophageal pacing enables detection of preclinical signs of lowered coronary reserve. PMID- 15821702 TI - [Structural and functional characteristics of cardiovascular system in elderly patients with borderline hypertension]. AB - Structural and functional state of the heart and vessels was studied in elderly patients with borderline hypertension. Remodeling in these patients was compared with that in patients of the same age with normal or higher blood pressure levels. Analysis revealed substantial contribution in remodeling of both age and hypertensive factors. Compared with subjects with normal blood pressure patients with hypertension had greater myocardial mass, higher frequency of left ventricular hypertrophy with cases of concentric hypertrophy and isolated hypertrophy of interventricular septum. Elderly patients with borderline hypertension had more pronounced disturbances of diastolic function. Overall characteristics of patients with borderline hypertension were similar to those found in the elderly with higher level of hypertension. Presence of borderline hypertension was associated with higher than age specific stiffness of vessels. Character and degree of remodeling of the heart and vessels in the elderly patients with borderline hypertension evidence for elevated risk of cardiovascular complications. PMID- 15821703 TI - [Nibentan -- a drug for pharmacological cardioversion of persistent atrial fibrillation]. AB - AIM: To assess efficacy of a potassium channel blocker nibentan for pharmacological cardioversion of persistent atrial fibrillation. Method. Nibentan (1% solution) was infused intravenously after 3 weeks of warfarin pretreatment to 31 patients with duration of atrial fibrillation from 4 weeks to 15 months. RESULTS: Sinus rhythm was restored in 24 patients. Mean dose of nibentan was 0.119+/-0.02 mg/kg. Proarrhythmic effects (paroxysmal or nonsustained polymorphic ventricular tachycardia) and substantial suppression of sinus rhythm automatism were registered in 3 and 3 patients, respectively. CONCLUSION: Nibentan is highly effective in persistent atrial fibrillation. It can be recommended for inhospital use in intensive care facilities. PMID- 15821704 TI - [Circadian dynamics of heart rate variability in patients with paroxysmal form of atrial fibrillation]. AB - Circadian dynamics of heart rate variability (HRV) was studied in patients with ischemic heart disease (IHD) with paroxysmal atrial fibrillation, with IHD and stable angina without clinically significant arrhythmia and healthy subjects of the same age. Special emphasis was placed on spontaneous changes of HRV parameters on short term ECG recordings obtained during unrestricted activity of a patient and dynamics of HRV parameters preceding nocturnal attacks of atrial fibrillation. The findings were decrease of total HRV, lowering throughout 24 hours of low and very low frequency power, and absence of circadian dynamics of characteristics of high frequency component of variability. HRV changes preceding attacks of atrial fibrillation were variable and in most cases both low and high frequency components were involved. This could evidence for participation in triggering of atrial fibrillation of both sympathetic and parasympathetic influences. PMID- 15821705 TI - [Dynamics of postinfarction left ventricular remodeling during long term use of perindopril, amiodarone, and beta-adrenoblockers]. AB - Perindopril was used in 287 patients with left ventricular ejection fraction < or =40% for 1 year after myocardial infarction either alone (n=53), or in combination with amiodarone (n=53), metoprolol (n=32), bisoprolol (n=32), carvedilol (n=32), atenolol (n=32), amiodarone and carvedilol (n=53). According to echocardiography data the use of combination of perindopril with metoprolol and especially carvedilol but not with amiodarone, bisoprolol, or atenolol, was associated with augmented anti-remodeling effect. The use of metoprolol, bisoprolol, and carvedilol but not of atenolol was associated with improvement of left ventricular systolic function. Amiodarone, metoprolol, bisoprolol, and atenolol, but not carvedilol diminished beneficial effect of perindopril on left ventricular diastolic function. The use of combination of perindopril with amiodarone or carvedilol was associated with less pronounced remodeling and most rapid and substantial improvement of left ventricular systolic and diastolic function. PMID- 15821706 TI - [Out of hospital pneumonia in patients with chronic heart failure: risk factors and frequency of development]. PMID- 15821707 TI - [Achievements and perspective of development of transplantology and artificial organs in Russia]. PMID- 15821708 TI - [Cardiovascular section of onco-surgery]. PMID- 15821710 TI - [Combination operations of valves and coronary arteries]. PMID- 15821711 TI - [Microvascular technique in coronary surgery]. PMID- 15821709 TI - [Problems of surgical treatment of patients with acute disorders of coronary circulation]. PMID- 15821712 TI - [Postoperative period in patients with cardiovascular pathology: empiric treatment or evidence based medicine?]. PMID- 15821713 TI - [Blood-loss minimalising technologies]. PMID- 15821714 TI - [Effect of granulocyte-macrophage colony stimulating factor on the function and regeneration of myocardium in rats with experimental infarction]. AB - To test the reparative capacity of stromal cells in myocardial infarction, rats were injected with granulocyte-monocyte-colony stimulating factor (GM-CSF) (leukomax), a cytokine known by its ability to raise a level of stromal cells in the blood, during first three days after coronary artery ligation. Only 10 of 17 rats (59%) survived 4 weeks in this group compared with 16 of 24 (67%) among rats not treated with leukomax. Echocardiographic and electromanometric studies showed that in both groups ventricular (LV) dilatation which developed during first hours after surgery persisted throughout 6-8 weeks and was combined with decreased ejection fraction and elevated LV end diastolic pressure. These alterations correlated with infarct size which varied from 0 to 28% of left ventricular weight in both groups. There were no statistically significant differences in functional and morphometric measurements between groups receiving and not receiving GM-CSF. However this result may be inconclusive due to small number of investigated animals and broad variation of ischemic zone size in each group. PMID- 15821715 TI - [Inotropic reactions of myocardium of patients with ischemic heart disease and rats during use of amiodarone]. AB - AIM: To study effect of amiodarone on inotropic reactions of myocardium of patients with ischemic heart disease and rats. METHODS AND RESULTS: Excitability of rat myocardium has been found to be higher than that of myocardium of patients. Sarcoplasmic reticulum of myocardium of rats has greater functional reserve, therefore rat muscle preparations respond to extrasystolic stimuli with postextrasystolic potentiation. Perfusion with a solution containing amiodarone caused diminution of amplitude of extrasystolic response of muscular strips both from rats and patients. Amiodarone augmented ability of sarcoplasmic reticulum to accumulate Ca ions as it prevented decrease of postextrasystolic contraction of human myocardium and brought about postextrasystolic potentiation of muscular strips of rat myocardium. CONCLUSION: Amiodarone affected intracellular homeostasis of calcium ions in cardiomyocytes of both rats and patients. Amiodarone lowered myocardial excitability and facilitated ability of cardiomyocyte sarcoplasmic reticulum to accumulate calcium ions. PMID- 15821716 TI - [Clinical value of the use of magnesium orotate in adolescents with syndrome of cardiac connective tissue dysplasia]. AB - According to results of clinical and instrumental investigation magnesium orotate (50 mg/day during first week and 25 mg/day thereafter) was found to be effective therapy of children with syndrome of cardiac connective tissue dysplasia (mainly with mitral valve prolapse and anomalous chordae tendineae). PMID- 15821717 TI - [A comparative assessment of valsartan and lisinopril effects on systolic and diastolic function of the left ventricular in patients with acute Q-infarction]. PMID- 15821718 TI - [Prevalence of hypertension among gas industry employees in Far North]. PMID- 15821722 TI - [Changing views on the place of drugs in the treatment of chronic heart failure]. PMID- 15821723 TI - [Infectious endocarditis: resistance to therapy and methods of its overcoming]. PMID- 15821724 TI - [Problems of self and mutual help in cardiovascular diseases and ways of its solution]. PMID- 15821725 TI - Gut microorganisms, mammalian metabolism and personalized health care. AB - The mammalian gut microbiota interact extensively with the host through metabolic exchange and co-metabolism of substrates. Such metabolome-metabolome interactions are poorly understood, but might be implicated in the aetiology of many human diseases. In this paper, we assess the importance of the gut microbiota in influencing the disposition, fate and toxicity of drugs in the host, and conclude that appropriate consideration of individual human gut microbial activities will be a necessary part of future personalized health-care paradigms. PMID- 15821727 TI - Ashbya gossypii: a model for fungal developmental biology. AB - Ashbya gossypii is a riboflavin-overproducing filamentous fungus that is closely related to unicellular yeasts such as Saccharomyces cerevisiae. With its close ties to yeast and the ease of genetic manipulation in this fungal species, A. gossypii is well suited as a model to elucidate the regulatory networks that govern the functional differences between filamentous growth and yeast growth, especially now that the A. gossypii genome sequence has been completed. Understanding these networks could be relevant to related dimorphic yeasts such as the human fungal pathogen Candida albicans, in which a switch in morphology from the yeast to the filamentous form in response to specific environmental stimuli is important for virulence. PMID- 15821726 TI - Bacterial cytotoxins: targeting eukaryotic switches. AB - Many bacterial cytotoxins act on eukaryotic cells by targeting the regulators that are involved in controlling the cytoskeleton or by directly modifying actin, with members of the Rho GTPase family being particularly important targets. The actin cytoskeleton, and especially the GTPase 'molecular switches' that are involved in its control, have crucial functions in innate and adaptive immunity, and have pivotal roles in the biology of infection. In this review, we briefly discuss the role of the actin cytoskeleton and the Rho GTPases in host-pathogen interactions, and review the mode of actions of bacterial protein toxins that target these components. PMID- 15821728 TI - Enzyme family-specific and activity-based screening of chemical libraries using enzyme microarrays. AB - The potential of protein microarrays in high-throughput screening (HTS) still remains largely unfulfilled, essentially because of the difficulty of extracting meaningful, quantitative data from such experiments. In the particular case of enzyme microarrays, low-molecular-weight fluorescent affinity labels (FALs) can function as ideally suited activity probes of the microarrayed enzymes. FALs form covalent bonds with enzymes in an activity-dependent manner and therefore can be used to characterize enzyme activity at each enzyme's address, as predetermined by the microarraying process. Relying on this principle, we introduce herein thematic enzyme microarrays (TEMA). In a kinetic setup we used TEMAs to determine the full set of kinetic constants and the reaction mechanism between the microarrayed enzymes (the theme of the microarray) and a family-wide FAL. Based on this kinetic understanding, in an HTS setup we established the practical and theoretical methodology for quantitative, multiplexed determination of the inhibition profile of compounds from a chemical library against each microarrayed enzyme. Finally, in a validation setup, K(i)(app) values and inhibitor profiles were confirmed and refined. PMID- 15821729 TI - Construction of lycopene-overproducing E. coli strains by combining systematic and combinatorial gene knockout targets. AB - Identification of genes that affect the product accumulation phenotype of recombinant strains is an important problem in industrial strain construction and a central tenet of metabolic engineering. We have used systematic (model-based) and combinatorial (transposon-based) methods to identify gene knockout targets that increase lycopene biosynthesis in strains of Escherichia coli. We show that these two search strategies yield two distinct gene sets, which affect product synthesis either through an increase in precursor availability or through (largely unknown) kinetic or regulatory mechanisms, respectively. Exhaustive exploration of all possible combinations of the above gene sets yielded a unique set of 64 knockout strains spanning the metabolic landscape of systematic and combinatorial gene knockout targets. This included a global maximum strain exhibiting an 8.5-fold product increase over recombinant K12 wild type and a twofold increase over the engineered parental strain. These results were further validated in controlled culture conditions. PMID- 15821730 TI - Midbrain dopamine and prefrontal function in humans: interaction and modulation by COMT genotype. AB - Using multimodal neuroimaging in humans, we demonstrate specific interactions between prefrontal activity and midbrain dopaminergic synthesis. A common V(108/158)M substitution in the gene for catecholamine-O-methyltransferase (COMT), an important enzyme regulating prefrontal dopamine turnover, predicted reduced dopamine synthesis in midbrain and qualitatively affected the interaction with prefrontal cortex. These data implicate a dopaminergic tuning mechanism in prefrontal cortex and suggest a systems-level mechanism for cognitive and neuropsychiatric associations with COMT. PMID- 15821731 TI - EphrinB-EphB signalling regulates clathrin-mediated endocytosis through tyrosine phosphorylation of synaptojanin 1. AB - Recent studies show that Eph receptors act mainly through the regulation of actin reorganization. Here, we show a novel mode of action for EphB receptors. We identify synaptojanin 1 - a phosphatidylinositol 5'-phosphatase that is involved in clathrin-mediated endocytosis - as a physiological substrate for EphB2. EphB2 causes tyrosine phosphorylation in the proline-rich domain of synaptojanin 1, and inhibits both the interaction with endophilin and the 5'-phosphatase activity of synaptojanin 1. Treatment with the EphB ligand, ephrinB2, elevates the cellular level of phosphatidylinositol 4,5-bisphosphate and promotes transferrin uptake. A kinase inactive mutant of EphB2 and a phosphorylation site mutant of synaptojanin 1 both neutralize the increase of transferrin uptake after ephrinB2 treatment. These mutants also inhibit AMPA glutamate receptor endocytosis in hippocampal neurons. Interestingly, incorporated transferrin does not reach endosomes, suggesting dual effects of EphB signalling on the early and late phases of clathrin-mediated endocytosis. Our results indicate that ephrinB-EphB signalling regulates clathrin-mediated endocytosis in various cellular contexts by influencing protein interactions and phosphoinositide turnover through tyrosine phosphorylation of synaptojanin 1. PMID- 15821732 TI - Actin and Arf1-dependent recruitment of a cortactin-dynamin complex to the Golgi regulates post-Golgi transport. AB - Cortactin is an actin-binding protein that has recently been implicated in endocytosis. It binds directly to dynamin-2 (Dyn2), a large GTPase that mediates the formation of vesicles from the plasma membrane and the Golgi. Here we show that cortactin associates with the Golgi to regulate the actin- and Dyn2 dependent transport of cargo. Cortactin antibodies stain the Golgi apparatus, labelling peripheral buds and vesicles that are associated with the cisternae. Notably, in vitro or intact-cell experiments show that activation of Arf1 mediates the recruitment of actin, cortactin and Dyn2 to Golgi membranes. Furthermore, selective disruption of the cortactin-Dyn2 interaction significantly reduces the levels of Dyn2 at the Golgi and blocks the transit of nascent proteins from the trans-Golgi network, resulting in swollen and distended cisternae. These findings support the idea of an Arf1-activated recruitment of an actin, cortactin and Dyn2 complex that is essential for Golgi function. PMID- 15821733 TI - Roberts syndrome is caused by mutations in ESCO2, a human homolog of yeast ECO1 that is essential for the establishment of sister chromatid cohesion. AB - Roberts syndrome is an autosomal recessive disorder characterized by craniofacial anomalies, tetraphocomelia and loss of cohesion at heterochromatic regions of centromeres and the Y chromosome. We identified mutations in a new human gene, ESCO2, associated with Roberts syndrome in 15 kindreds. The ESCO2 protein product is a member of a conserved protein family that is required for the establishment of sister chromatid cohesion during S phase and has putative acetyltransferase activity. PMID- 15821734 TI - MYCN haploinsufficiency is associated with reduced brain size and intestinal atresias in Feingold syndrome. AB - Feingold syndrome is characterized by variable combinations of esophageal and duodenal atresias, microcephaly, learning disability, syndactyly and cardiac defect. We show here that heterozygous mutations in the gene MYCN are present in Feingold syndrome. All mutations are predicted to disrupt both the full-length protein and a new shortened MYCN isoform, suggesting that multiple aspects of early embryogenesis and postnatal brain growth in humans are tightly regulated by MYCN dosage. PMID- 15821735 TI - Epistasis analysis with global transcriptional phenotypes. AB - Classical epistasis analysis can determine the order of function of genes in pathways using morphological, biochemical and other phenotypes. It requires knowledge of the pathway's phenotypic output and a variety of experimental expertise and so is unsuitable for genome-scale analysis. Here we used microarray profiles of mutants as phenotypes for epistasis analysis. Considering genes that regulate activity of protein kinase A in Dictyostelium, we identified known and unknown epistatic relationships and reconstructed a genetic network with microarray phenotypes alone. This work shows that microarray data can provide a uniform, quantitative tool for large-scale genetic network analysis. PMID- 15821736 TI - MHC2TA is associated with differential MHC molecule expression and susceptibility to rheumatoid arthritis, multiple sclerosis and myocardial infarction. AB - Antigen presentation to T cells by MHC molecules is essential for adaptive immune responses. To determine the exact position of a gene affecting expression of MHC molecules, we finely mapped a previously defined rat quantitative trait locus regulating MHC class II on microglia in an advanced intercross line. We identified a small interval including the gene MHC class II transactivator (Mhc2ta) and, using a map over six inbred strains combined with gene sequencing and expression analysis, two conserved Mhc2ta haplotypes segregating with MHC class II levels. In humans, a -168A --> G polymorphism in the type III promoter of the MHC class II transactivator (MHC2TA) was associated with increased susceptibility to rheumatoid arthritis, multiple sclerosis and myocardial infarction, as well as lower expression of MHC2TA after stimulation of leukocytes with interferon-gamma. We conclude that polymorphisms in Mhc2ta and MHC2TA result in differential MHC molecule expression and are associated with susceptibility to common complex diseases with inflammatory components. PMID- 15821737 TI - Intestinal immune homeostasis is regulated by the crosstalk between epithelial cells and dendritic cells. AB - The control of damaging inflammation by the mucosal immune system in response to commensal and harmful ingested bacteria is unknown. Here we show epithelial cells conditioned mucosal dendritic cells through the constitutive release of thymic stromal lymphopoietin and other mediators, resulting in the induction of 'noninflammatory' dendritic cells. Epithelial cell-conditioned dendritic cells released interleukins 10 and 6 but not interleukin 12, and they promoted the polarization of T cells toward a 'classical' noninflammatory T helper type 2 response, even after exposure to a T helper type 1-inducing pathogen. This control of immune responses seemed to be lost in patients with Crohn disease. Thus, the intimate interplay between intestinal epithelial cells and dendritic cells may help to maintain gut immune homeostasis. PMID- 15821738 TI - T cell costimulation by chemokine receptors. AB - Signals mediated by chemokine receptors may compete with T cell receptor stop signals and determine the duration of T cell-antigen-presenting cell interactions. Here we show that during T cell stimulation by antigen-presenting cells, T cell chemokine receptors coupled to G(q) and/or G(11) protein were recruited to the immunological synapse by a G(i)-independent mechanism. When chemokine receptors were sequestered at the immunological synapse, T cells became insensitive to chemotactic gradients, formed more stable conjugates and finally responded with enhanced proliferation and cytokine production. We suggest that chemokine receptor trapping at the immunological synapse enhances T cell activation by improving T cell-antigen-presenting cell attraction and impeding the 'distraction' of successfully engaged T cells by other chemokine sources. PMID- 15821739 TI - Inhibition of the NKp30 activating receptor by pp65 of human cytomegalovirus. AB - Human cytomegalovirus, a chief pathogen in immunocompromised people, can persist in a healthy immunocompetent host throughout life without being eliminated by the immune system. Here we show that pp65, the main tegument protein of human cytomegalovirus, inhibited natural killer cell cytotoxicity by an interaction with the activating receptor NKp30. This interaction was direct and specific, leading to dissociation of the linked CD3zeta from NKp30 and, consequently, to reduced killing. Thus, pp65 is a ligand for the NKp30 receptor and demonstrates a unique mechanism by which an intracellular viral protein causes general suppression of natural killer cell cytotoxicity by specific interaction with an activating receptor. PMID- 15821741 TI - Chiral selectivity in the charge-transfer bleaching of single-walled carbon nanotube spectra. AB - Chiral selective reactivity and redox chemistry of carbon nanotubes are two emerging fields of nanoscience. These areas hold strong promise for producing methods for isolating nanotubes into pure samples of a single electronic type, and for reversible doping of nanotubes for electronics applications. Here, we study the selective reactivity of single-walled carbon nanotubes with organic acceptor molecules. We observe spectral bleaching of the nanotube electronic transitions consistent with an electron-transfer reaction occurring from the nanotubes to the organic acceptors. The reaction kinetics are found to have a strong chiral dependence, with rates being slowest for large-bandgap species and increasing for smaller-bandgap nanotubes. The chiral-dependent kinetics can be tuned to effectively freeze the reacted spectra at a fixed chiral distribution. Such tunable redox chemistry may be important for future applications in reversible non-covalent modification of nanotube electronic properties and in chiral selective separations. PMID- 15821740 TI - Unconventional topology of self peptide-major histocompatibility complex binding by a human autoimmune T cell receptor. AB - Autoimmune diseases are caused by self-reactive lymphocytes that have escaped deletion. Here we have determined the structure of the trimolecular complex for a T cell receptor (TCR) from a patient with multiple sclerosis that causes autoimmunity in transgenic mice. The structure showed a TCR topology notably different from that of antimicrobial TCRs. Rather than being centered on the peptide-major histocompatibility complex, this TCR contacted only the N-terminal peptide segment and made asymmetrical interactions with the major histocompatibility complex helices. The interaction was dominated by the hypervariable complementarity-determining region 3 loops, indicating that unconventional topologies are possible because of the unique complementarity determining region 3 sequences created during rearrangement. This topology reduces the interaction surface with peptide and alters the geometry for CD4 association. We propose that unusual TCR-binding properties can permit autoreactive T cells to escape deletion. PMID- 15821742 TI - Analysis of DNA supercoil induction by FtsK indicates translocation without groove-tracking. AB - FtsK is a bacterial protein that translocates DNA in order to transport chromosomes within the cell. During translocation, DNA's double-helical structure might cause a relative rotation between FtsK and the DNA. We used a single molecule technique to quantify this rotation by observing the supercoils induced into the DNA during translocation of an FtsK complex. We find that FtsK induces approximately 0.07 supercoils per DNA helical pitch traveled. This rate indicates that FtsK does not track along DNA's groove, but it is consistent with our previous estimate of FtsK's step size. We show that this rate of supercoil induction is markedly near to the ideal value that would minimize in vivo disturbance to the chromosomal supercoil density, suggesting an origin for the unusual rotational behavior of FtsK. PMID- 15821743 TI - The orphan nuclear receptor RORalpha regulates circadian transcription of the mammalian core-clock Bmal1. AB - The PAS (PER-ARNT-SIM) helix-loop-helix transcription factor BMAL1 (also known as MOP3) is an essential component of the circadian pacemaker in mammals. Here we show that the retinoic acid receptor-related orphan receptor RORalpha (NR1F1) directly activates transcription of Bmal1 through two conserved RORalpha response elements that are required for cell-autonomous transcriptional oscillation of Bmal1 mRNA. Positive involvement of RORalpha in generation of the Bmal1 circadian oscillation was verified by behavioral analyses of RORalpha-deficient staggerer mice that showed aberrant locomotor activity and unstable rhythmicity. In cultured cells, loss of endogenous RORalpha protein resulted in a dampened circadian rhythm of Bmal1 transcription, further indicating that RORalpha is a functional component of the cell-autonomous core circadian clock. These results indicate that RORalpha acts to promote Bmal1 transcription, thereby maintaining a robust circadian rhythm. PMID- 15821744 TI - Ribosomal protein L30 is a component of the UGA-selenocysteine recoding machinery in eukaryotes. AB - The translational recoding of UGA as selenocysteine (Sec) is directed by a SECIS element in the 3' untranslated region (UTR) of eukaryotic selenoprotein mRNAs. The selenocysteine insertion sequence (SECIS) contains two essential tandem sheared G.A pairs that bind SECIS-binding protein 2 (SBP2), which recruits a selenocysteine-specific elongation factor and Sec-tRNA(Sec) to the ribosome. Here we show that ribosomal protein L30 is a component of the eukaryotic selenocysteine recoding machinery. L30 binds SECIS elements in vitro and in vivo, stimulates UGA recoding in transfected cells and competes with SBP2 for SECIS binding. Magnesium, known to induce a kink-turn in RNAs that contain two tandem G.A pairs, decreases the SBP2-SECIS complex in favor of the L30-SECIS interaction. We propose a model in which SBP2 and L30 carry out different functions in the UGA recoding mechanism, with the SECIS acting as a molecular switch upon protein binding. PMID- 15821745 TI - Hemifusion in SNARE-mediated membrane fusion. AB - SNAREs are essential for intracellular membrane fusion. Using EPR, we determined the structure of the transmembrane domain (TMD) of the vesicle (v)-SNARE Snc2p involved in trafficking in yeast. Structural features of the TMD were used to design a v-SNARE mutant in which about half of the TMD was deleted. Liposomes containing this mutant induced outer leaflet mixing but not inner leaflet mixing when incubated with liposomes containing target membrane (t)-SNAREs. Hemifusion was also detected with wild-type SNAREs when low protein concentrations were reconstituted. Thus, these results show that SNARE-mediated fusion can transit through a hemifusion intermediate. PMID- 15821746 TI - Lineage infidelity of epithelial ovarian cancers is controlled by HOX genes that specify regional identity in the reproductive tract. AB - Although epithelial ovarian cancers (EOCs) have been thought to arise from the simple epithelium lining the ovarian surface or inclusion cysts, the major subtypes of EOCs show morphologic features that resemble those of the mullerian duct-derived epithelia of the reproductive tract. We found that HOX genes, which normally regulate mullerian duct differentiation, are not expressed in normal ovarian surface epithelium (OSE), but are expressed in different EOC subtypes according to the pattern of mullerian-like differentiation of these cancers. Ectopic expression of Hoxa9 in tumorigenic mouse OSE cells gave rise to papillary tumors resembling serous EOCs. In contrast, Hoxa10 and Hoxa11 induced morphogenesis of endometrioid-like and mucinous-like EOCs, respectively. Hoxa7 showed no lineage specificity, but promoted the abilities of Hoxa9, Hoxa10 and Hoxa11 to induce differentiation along their respective pathways. Therefore, inappropriate activation of a molecular program that controls patterning of the reproductive tract could explain the morphologic heterogeneity of EOCs and their assumption of mullerian-like features. PMID- 15821747 TI - In vivo imaging of axonal degeneration and regeneration in the injured spinal cord. AB - The poor response of central axons to transection underlies the bleak prognosis following spinal cord injury. Here, we monitor individual fluorescent axons in the spinal cords of living transgenic mice over several days after spinal injury. We find that within 30 min after trauma, axons die back hundreds of micrometers. This acute form of axonal degeneration is similar in mechanism to the more delayed Wallerian degeneration of the disconnected distal axon, but acute degeneration affects the proximal and distal axon ends equally. In vivo imaging further shows that many axons attempt regeneration within 6-24 h after lesion. This growth response, although robust, seems to fail as a result of the inability of axons to navigate in the proper direction. These results suggest that time lapse imaging of spinal cord injury may provide a powerful analytical tool for assessing the pathogenesis of spinal cord injury and for evaluating therapies that enhance regeneration. PMID- 15821748 TI - An essential function for NBS1 in the prevention of ataxia and cerebellar defects. AB - Nijmegen breakage syndrome (NBS), ataxia telangiectasia and ataxia telangiectasia like disorder (ATLD) show overlapping phenotypes such as growth retardation, microcephaly, cerebellar developmental defects and ataxia. However, the molecular pathogenesis of these neurological defects remains elusive. Here we show that inactivation of the Nbn gene (also known as Nbs1) in mouse neural tissues results in a combination of the neurological anomalies characteristic of NBS, ataxia telangiectasia and ATLD, including microcephaly, growth retardation, cerebellar defects and ataxia. Loss of Nbn causes proliferation arrest of granule cell progenitors and apoptosis of postmitotic neurons in the cerebellum. Furthermore, Nbn-deficient neuroprogenitors show proliferation defects (but not increased apoptosis) and contain more chromosomal breaks, which are accompanied by ataxia telangiectasia mutated protein (ATM)-mediated p53 activation. Notably, depletion of p53 substantially rescues the neurological defects of Nbn mutant mice. This study gives insight into the physiological function of NBS1 (the Nbn gene product) and the function of the DNA damage response in the neurological anomalies of NBS, ataxia telangiectasia and ATLD. PMID- 15821749 TI - Deletion of Gab1 in the liver leads to enhanced glucose tolerance and improved hepatic insulin action. AB - Insulin receptor substrate-1 (IRS-1) and IRS-2 are known to transduce and amplify signals emanating from the insulin receptor. Here we show that Grb2-associated binder 1 (Gab1), despite its structural similarity to IRS proteins, is a negative modulator of hepatic insulin action. Liver-specific Gab1 knockout (LGKO) mice showed enhanced hepatic insulin sensitivity with reduced glycemia and improved glucose tolerance. In LGKO liver, basal and insulin-stimulated tyrosine phosphorylation of IRS-1 and IRS-2 was elevated, accompanied by enhanced Akt/PKB activation. Conversely, Erk activation by insulin was suppressed in LGKO liver, leading to defective IRS-1 Ser612 phosphorylation. Thus, Gab1 acts to attenuate, through promotion of the Erk pathway, insulin-elicited signals flowing through IRS and Akt proteins, which represents a novel balancing mechanism for control of insulin signal strength in the liver. PMID- 15821750 TI - Neuroprotective activity of the mGluR5 antagonists MPEP and MTEP against acute excitotoxicity differs and does not reflect actions at mGluR5 receptors. AB - 1 Neuroprotection has been reported after either activation or blockade of the group I metabotropic glutamate receptor subtype 5 (mGluR5). However, some recent evidence suggests that protection provided by mGluR5 antagonists may reflect their ability to inhibit N-methyl-D-aspartate (NMDA) receptor activity. 2 Here, in both rat and mouse cortical neurons, we compare the neuroprotective actions of two mGluR5 antagonists: 2-methyl-6-(phenylethynyl)-pyridine (MPEP), which has been commonly used and 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP), a more recently developed compound believed to have greater mGluR5 selectivity. We have previously shown that MPEP directly reduces single-channel NMDA receptor open time at the same concentrations (20 microM or greater) that show neuroprotection, whereas MPEP antagonizes mGluR5 agonist ((RS)-2-chloro-5 hydroxyphenylglycine (CHPG))-induced changes in inositol phosphates (IP) at concentrations as low as 0.2 microM. 3 In the present studies, MTEP significantly inhibited CHPG-mediated IP hydrolysis at concentrations as low as 0.02 microM. In contrast to MPEP, which significantly reduced glutamate- or NMDA-mediated cell death in primary rat neuronal cultures at a concentration of 20 microM, small neuroprotective effects were observed with MTEP only at a concentration of 200 microM. Neither MPEP- nor MTEP-mediated mGluR5 inhibition had any effect on etoposide-induced apoptotic cell death. In rat cortical neurons, the neuroprotective effects of MTEP at very high concentrations, like those of MPEP, reflect ability to directly reduce NMDA receptor peak and steady-state currents. 4 We also compared the effects of MPEP and MTEP in primary cortical neuronal cultures from parental and mGluR5 knockout mice. Both agents were neuroprotective, at high concentrations in normal as well as in the knockout cultures. In contrast to rat cortical neurons, neither MPEP nor MTEP appears to directly alter NMDA receptor activity. 5 Combined, these studies support the conclusion that MTEP has greater mGluR5 selectivity than MPEP, and that neuroprotection provided by either antagonist in neuronal cultures does not reflect inhibition of mGluR5 receptors. PMID- 15821752 TI - Activation of calcineurin in human failing heart ventricle by endothelin-1, angiotensin II and urotensin II. AB - 1 The calcineurin (CaN) enzyme-transcriptional pathway is critically involved in hypertrophy of heart muscle in some animal models. Currently there is no information concerning the regulation of CaN activation by endogenous agonists in human heart. 2 Human right ventricular trabeculae from explanted human (14 male/2 female) failing hearts were set up in a tissue bath and electrically paced at 1 Hz and incubated with or without 100 nM endothelin-1 (ET-1), 10 M, angiotensin-II (Ang II) or 20 nM human urotensin-II (hUII) for 30 min. Tissues from four patients were incubated with 200 nM tacrolimus (FK506) for 30 min and then incubated in the presence or absence of ET-1 for a further 30 min. 3 ET-1 increased contractile force in all 13 patients (P<0.001). Ang II and hUII increased contractile force in three out of eight and four out of 10 patients but overall nonsignificantly (P>0.1). FK506 had no effect on contractile force (P=0.12). 4 ET-1, Ang II and hUII increased calcineurin activity by 32, 71 and 15%, respectively, while FK506 reduced activity by 34%. ET-1 in the presence of FK506 did not restore calcineurin activity (P=0.1). 5 There was no relationship between basal CaN activity and expression levels in the right ventricle. Increased levels of free phosphate were detected in ventricular homogenates that were incubated with PKC(epsilon) compared to samples incubated without PKC(epsilon). 6 Endogenous cardiostimulants which activate G(alpha)q-coupled receptors increase the activity of calcineurin in human heart following acute (30 min) exposure. PKC may contribute to this effect by increasing levels of phosphorylated calcineurin substrate. PMID- 15821751 TI - The effects of Delta9-tetrahydrocannabinol in rat mesenteric vasculature, and its interactions with the endocannabinoid anandamide. AB - 1 Delta9-tetrahydrocannabinol (THC) produces varying effects in mesenteric arteries: vasorelaxation (third-order branches, G3), modest vasorelaxation (G2), no effect (G1) and vasoconstriction (the superior mesenteric artery, G0). 2 In G3, vasorelaxation to THC was inhibited by pertussis toxin, but was unaffected by the CB1 receptor antagonist, AM251 (1 microM), incubation with the TRPV1 receptor agonist capsaicin (10 microM, 1 h), the TRPV1 receptor antagonist capsazepine (10 microM) or de-endothelialisation. 3 In G3, vasorelaxation to THC was inhibited by high K+ buffer, and by the following K+ channel inhibitors: charybdotoxin (100 nM), apamin (500 nM) and barium chloride (30 microM), but not by 4-aminopyridine, glibenclamide or tertiapin. 4 In G3, THC (10 and 100 microM) inhibited the contractile response to Ca2+ in a Ca2+-free, high potassium buffer, indicating that THC blocks Ca2+ influx. 5 In G0, the vasoconstrictor responses to THC were inhibited by de-endothelialisation and SR141716A (100 nM), but not by the endothelin (ET(A)) receptor antagonist FR139317 (1 microM).THC (1 and 10 microM) antagonised vasorelaxation to anandamide in G3 but not G0. THC did not antagonise the noncannabinoid verapamil, capsaicin or the CB1 receptor agonist CP55,940. 6 THC (10 and 100 microM) inhibited endothelium-derived relaxing factor (EDHF) mediated responses to carbachol in a manner similar to the gap junction inhibitor 18alpha-glycyrrhetinic acid. 7 These data show that THC causes vasorelaxation through activation of K+ channels and inhibition of Ca2+ channels, and this involves non-CB1, non-TRPV1 but G-protein-coupled receptors. In G0, THC does not cause relaxation and at high concentrations causes contractions. Importantly, THC antagonises the effects of anandamide, possibly through inhibition of EDHF activity. PMID- 15821753 TI - Activation of cannabinoid CB2 receptor negatively regulates IL-12p40 production in murine macrophages: role of IL-10 and ERK1/2 kinase signaling. AB - 1 Cannabinoid (CB) receptor agonists have potential utility as anti-inflammatory drugs for the treatment of many disease conditions. In the present study, we investigated the effects of the synthetic CB(2) ligand, JWH-133 on the production of interleukins (ILs), IL-12 and IL-10 by lipopolyssacharide (LPS) or Theiler's virus (TMEV)-activated macrophages. 2 JWH-133 evoked a concentration-related inhibition (10 nM-5 microM) of LPS/IFN-gamma induced IL-12p40 release. The effect of JWH-133 (100 nM) was significantly blocked by the CB2 antagonist SR-144528 (1 microM). Macrophages infected with TMEV increased IL-12p40 production and activation of CB2 receptors by JWH-133 (100 nM) inhibited it. 3 The inhibitory effect of JWH-133 (100 nM) on IL-12p40 production may involve extracellular regulated kinase (ERK1/2) signaling: (i) JWH-133 induced a greater and sustained activation of ERK1/2 kinase in comparison with the level of activation observed following LPS; (ii) the inhibition of ERK1/2 by the specific inhibitor PD98059 increased LPS-induced IL-12p40 production in the presence or absence of JWH-133 suggesting a negative regulation of ERK pathway on IL-12p40 biosynthesis. 4 Activation of CB2 receptors by JWH-133 (10 nM-5 microM) enhanced IL-10 release by LPS/IFN-gamma-activated macrophages and addition of SR144558 (1 microM) totally blocked the effect of JWH (100 nM). 5 Inhibition of ERK by PD98059 significantly suppressed IL-10 production by LPS-activated macrophages. Endogenous IL-10 plays a modulatory role in IL-12 production. Blocking IL-10 with neutralizing antibody resulted in increased IL-12p40 secretion by LPS-activated macrophages in the absence or presence of JWH-133. In contrast, the addition of exogenous mIL-10 reduced the secretion of IL-12p40 in response to LPS. PMID- 15821754 TI - Novel type of Gq/11 protein-coupled neurosteroid receptor sensitive to endocrine disrupting chemicals in mast cell line (RBL-2H3). AB - 1 Agonistic neurosteroids, including pregnenolone, dehydroepiandrosterone and its sulfate (DHEAS), caused rapid degranulation in measurements of beta hexosaminidase (beta-HEX) release from a mast cell line, RBL-2H3. This degranulation was blocked by BSA-conjugated progesterone (PROG-BSA) or 17beta estradiol, both of which are antagonistic neurosteroids. 2 DHEAS-induced beta-HEX release was blocked by U-73122 or xestospongin C, but not by PTX or EGTA. DHEAS induced beta-HEX release was also abolished by G(q/11)-AS, but not by G(q/11)-MS. Pharmacological analyses revealed that the neurosteroids stimulated a putative membrane receptor through activation of the novel G(q/11) and phospholipase C. 3 While representative endocrine-disrupting chemicals (EDCs) did not show any degranulation or nocifensive actions by themselves, they blocked the DHEAS induced degranulation. 4 The binding of a PROG-BSA-fluorescein isothiocyanate conjugate (PROG-BSA-FITC) to cells was inhibited by neurosteroids and EDCs. 5 In the algogenic-induced biting and licking responses test, DHEAS caused agonistic nocifensive actions in a dose-dependent manner between 1 and 10 fmol (i.pl.). DHEAS-induced nocifensive actions were abolished by PROG-BSA or nonylphenol. 6 Taken together, these results suggest that a G(q/11)-coupled neurosteroid receptor may regulate the neuroimmunological activity related to sensory stimulation and that some EDCs have antagonistic actions for this receptor. PMID- 15821755 TI - Prostaglandin E2-induced modification of tetrodotoxin-resistant Na+ currents involves activation of both EP2 and EP4 receptors in neonatal rat nodose ganglion neurones. AB - 1 The aim of the present study was to investigate which EP receptor subtypes (EP1 EP4) act predominantly on the modification of the tetrodotoxin-resistant Na+ current (I(NaR)) in acutely isolated neonatal rat nodose ganglion (NG) neurones. 2 Of the four EP receptor agonists ranging from 0.01 to 10 muM, the EP2 receptor agonist (ONO-AE1-259, 0.1-10 microM) and the EP4 receptor agonist (ONO-AE1-329, 1 microM) significantly increased peak I(NaR). The responses were associated with a hyperpolarizing shift in the activation curve. 3 Neither the EP1 receptor agonist ONO-DI-004 nor the EP3 receptor agonist ONO-AE-248 significantly modified the properties of I(NaR). 4 In PGE2 applications ranging from 0.01 to 10 microM, 1 microM PGE2 produced a maximal increase in the peak I(NaR) amplitude. The PGE2 (1 microM)-induced increase in the GV(1/2) baseline (% change in G at baseline V(1/2)) was significantly attenuated by either intracellular application of the PKA inhibitor PKI or extracellular application of the protein kinase C inhibitor staurosporine (1 microM). However, the slope factor k was not significantly altered by PGE2 applications at 0.01-10 microM. In addition, the hyperpolarizing shift of V(1/2) by PGE2 was not significantly altered by either PKI or staurosporine. 5 In other series of experiments, reverse transcription-polymerase chain reaction (RT-PCR) of mRNA from nodose ganglia indicated that all four EP receptors were present. 6 The NG contained many neuronal cell bodies (diameter <30 microm) with intense or moderate EP2, EP3, and EP4 receptor immunoreactivities. 7 These results suggest that the PGE2-induced modification of I(NaR) is mainly mediated by activation of both EP2 and EP4 receptors. PMID- 15821756 TI - Annexin-1 peptide Anx-1(2-26) protects adult rat cardiac myocytes from cellular injury induced by simulated ischaemia. AB - 1 The anti-inflammatory properties of annexin-1 peptides have been largely ascribed to their powerful antineutrophil actions in vivo. We have recently reported that the N-terminal fragment of annexin-1, Anx-1(2-26), preserves contractile function of cardiac muscle in vitro. The aim of the present study was to determine if Anx-1(2-26) elicits protective actions specifically on the cardiac myocyte (in the absence of neutrophils), using a model of metabolic inhibition to simulate ischaemia. 2 Metabolic inhibition of cardiac myocytes (4 h incubation at 37 degrees C in HEPES-containing buffer supplemented with 2-deoxy-D glucose, D,L-lactic acid and pH adjusted to 6.5) followed by 2.5 h recovery in normal medium markedly increased creatine kinase (CK) and lactate dehydrogenase (LDH) levels by 179+/-39 and 26+/-7 IU L(-1) (both n=40, P<0.001), respectively. However, cellular injury was significantly decreased when Anx-1(2-26) (0.3 microM) was present during metabolic inhibition, CK by 74+/-10% and LDH by 71+/ 6% (both n=31, P<0.001), respectively. 3 Boc 2 (10 microM), a nonselective formyl peptide receptor antagonist, present during metabolic inhibition, abolished the cardioprotective effect of Anx-1(2-26). 4 Addition of chelerythrine (10 microM), 5-hydroxydecanoate (500 microM) or SB202190 (1 microM) during metabolic inhibition also abolished Anx-1(2-26)-induced cardioprotection. 5 Cellular injury induced by metabolic inhibition was also largely prevented when myocytes were incubated with Anx-1(2-26) for 5 min with 10 min recovery prior to the insult, or when Anx-1(2-26) was present only during the recovery period following drug-free metabolic inhibition. 6 In conclusion, the annexin-1 peptide Anx-1(2-26) potently prevents cardiac myocyte injury induced by metabolic inhibition, an action that was dependent at least in part on the activation of the formyl peptide receptor family of G-protein-coupled receptors, protein kinase C, p38 mitogen-activated protein kinase and ATP-sensitive potassium channels. PMID- 15821757 TI - Effects of bisindolylmaleimide PKC inhibitors on p90RSK activity in vitro and in adult ventricular myocytes. AB - 1 Bisindolylmaleimide inhibitors of protein kinase C (PKC), such as GF109203X and Ro31-8220, have been used to investigate the roles of PKC isoforms in many cellular processes in cardiac myocytes, but these agents may also inhibit p90RSK activity. 2 In in vitro kinase assays utilising 50 microM [ATP], GF109203X and Ro31-8220 inhibited p90RSK isoforms (IC50 values for inhibition of RSK1, RSK2 and RSK3, respectively, were 610, 310 and 120 nM for GF109203X, and 200, 36 and 5 nM for Ro31-8220) as well as classical and novel PKC isoforms (IC50 values for inhibition of PKCalpha and PKCepsilon, respectively, were 8 and 12 nM for GF109203X, and 4 and 8 nM for Ro31-8220). 3 At physiological [ATP] (5 mM), both GF109203X and Ro31-8220 exhibited reduced potency as inhibitors of RSK2, PKCalpha and PKCepsilon (IC50 values of 7400, 310 and 170 nM, respectively, for GF109203X, and 930, 150 and 140 nM, respectively, for Ro31-8220), with the latter agent retaining its relatively greater potency. 4 To determine the effects of GF109203X and Ro31-8220 on p90RSK activity in cultured adult rat ventricular myocytes (ARVM), phosphorylation of the eukaryotic elongation factor 2 kinase (eEF2K) at Ser366, a known p90RSK target, was used as the index of such activity. Adenoviral expression of a constitutively active form of mitogen-activated protein kinase (MAPK) or extracellular signal-regulated kinase (ERK) kinase 1 (MEK1) was used to induce PKC-independent p90RSK activation and downstream phosphorylation of eEF2K. 5 eEF2K phosphorylation was abolished by U0126 (1 microM), a selective inhibitor of MEK1, and was significantly reduced by GF109203X at > or =3 microM and by Ro31 8220 at > or =1 microM. At 1 microM, both agents inhibited PMA-induced PKC activity in ARVM. 6 These data show that GF109203X and Ro31-8220 inhibit various isoforms of PKC and p90RSK in vitro and in intact ARVM, with the former agent exhibiting relatively greater selectivity for PKC. PMID- 15821759 TI - 5-Lipoxygenase modulates colitis through the regulation of adhesion molecule expression and neutrophil migration. AB - Leukotrienes play a part in inflammatory response. The unique role of the enzyme 5-lipoxygenase (5-LO) in the production of leukotrienes makes it as therapeutic target for inflammatory conditions like inflammatory bowel disease (IBD). In the present study, by comparing the responses in wild-type mice (5-LOWT) and mice lacking the 5-lipoxygenase (5-LOKO), we investigated the role played by this enzyme in the development of experimental colitis. To address this question, we used an experimental model of colitis, induced by dinitrobenzene sulfonic acid (DNBS). When compared to DNBS-treated 5-LOWT mice, DNBS-treated 5-LOKO mice experienced a reduced rate of the extent and severity of the histological signs of colon injury. After administration of DNBS 5-LOWT mice showed hemorrhagic diarrhea, weight loss and large areas of necrosis in the mucosa of the colon. Neutrophil infiltration was associated with the expression of ICAM-1, VCAM-1, P selectin, E-selectin that were mainly localized around vessels. Absence of a functional 5-LO resulted in a significant reduction of all the above-described parameters. In particular, we have observed a significant reduction of: (i) the degree of colon injury, (ii) the rise in myeloperoxidase (MPO) activity (mucosa), (iii) the increase in staining (immunohistochemistry) for ICAM-1, VCAM-1, P selectin, E-selectin caused by DNBS in the colon. Similarly, the treatment of 5 LOWT with zileuton (50 mg/kg per os twice a day) resulted in a significant reduction of all the above-described parameters. In addition, in in vitro study a significantly reduced chemotactic response to IL-8 was observed in peripheral blood leukocytes from 5-LOKO in comparison to 5-LOWT polymorphonuclear leukocyte. Similar results were obtained when we analyzed the chemotactic response of 5-LOWT cell incubated for 15 min with zileuton (50 microg/ml). Taken together, our results clearly demonstrate that 5-LO modulates neutrophil infiltration in experimental colitis through the expression of adhesion molecules. PMID- 15821758 TI - The role of intracellular Ca2+ in the regulation of proteinase-activated receptor 2 mediated nuclear factor kappa B signalling in keratinocytes. AB - 1 In this study, we examined the role of Ca2+ in linking proteinase-activated receptor-2 (PAR2) to the nuclear factor kappa B (NFkappaB) pathway in a skin epithelial cell line NCTC2544 stably expressing PAR2 (clone G). 2 In clone G, PAR2-mediated NFkappaB luciferase reporter activity and NFkappaB DNA-binding activity was reduced by preincubation with BAPTA-AM but not BAPTA. Trypsin stimulation of inhibitory kappa B kinases, IKKalpha and IKKbeta, was also inhibited following pretreatment with BAPTA-AM. 3 BAPTA/AM also prevented PAR2 mediated IKKalpha activation in cultured primary human keratinocytes. 4 The effect of BAPTA-AM was also selective for the IKK/NFkappaB signalling axis; PAR2 coupling to ERK, or p38 MAP kinase was unaffected. 5 Pharmacological inhibition of the Ca2+-dependent regulatory protein calcineurin did not inhibit trypsin stimulated IKK activity or NFkappaB-DNA binding; however, inhibition of Ca2+ dependent protein kinase C isoforms or InsP3 formation using GF109203X or the phospholipase C inhibitor U73122, respectively, reduced both IKK activity and NFkappaB-DNA binding. 6 Mutation of PAR2 within the C-terminal to produce a mutant receptor, which does not couple to Ca2+ signalling, but is able to activate ERK, abrogated NFkappaB-DNA binding and IKK activity stimulated by trypsin. 7 These results suggest a predominant role for the InsP3/Ca2+ axis in the regulation of IKK signalling and NFkappaB transcriptional activation. PMID- 15821760 TI - Psychological distress of patients undergoing intensified conditioning with radioimmunotherapy prior to allogeneic stem cell transplantation. AB - This is a pilot study comparing the emotional distress of patients receiving an intensified conditioning regimen (radioimmunotherapy=RIT) with patients receiving conventional conditioning for allogeneic stem cell transplantation. In total, 53 patients (18 received RIT) were given two questionnaires designed to measure emotional distress (HADS, POMS) before starting conditioning (t1) and at discharge (t2). During the in-patient period, patients answered questions daily relating to physical distress, psychological distress, and how they were "coping with the situation". At t2, the transplant team assessed the manner in which the patients were coping. The data displayed no relevant differences with regard to emotional distress between the two groups, both at t1 and t2. For both groups, anxiety and vigor decreased and fatigue increased between t1 and t2. On average, perceived distress was higher for those patients being treated with RIT during the in-patient time, but the differences between both groups were significant only regarding physical distress during the recovery period. No difference was found for the transplant team's assessment. We hypothesize that an intensified conditioning regimen with RIT per se has only a small distressing effect on the patients' psyche during their stay at the hospital. Differences between both groups probably result from independent factors such as, for example, the patients' pre-existing health conditions. PMID- 15821761 TI - Mesenchymal stem cells rescue CD34+ cells from radiation-induced apoptosis and sustain hematopoietic reconstitution after coculture and cografting in lethally irradiated baboons: is autologous stem cell therapy in nuclear accident settings hype or reality? AB - Autologous stem cell therapy (ACT) has been proposed to prevent irradiated victims from bone marrow (BM) aplasia by grafting hematopoietic stem and progenitor cells (HSPCs) collected early after damage, provided that a functional graft of sufficient size could be produced ex vivo. To address this issue, we set up a baboon model of cell therapy in which autologous peripheral blood HSPCs collected before lethal total body irradiation were irradiated in vitro (2.5 Gy, D0 1 Gy) to mimic the cell damage, cultured in small numbers for a week in a serum-free medium in the presence of antiapoptotic cytokines and mesenchymal stem cells (MSCs) and then cografted. Our study shows that baboons cografted with expanded cells issued from 0.75 and 1 x 10(6)/kg irradiated CD34+ cells and MSCs (n=2) exhibited a stable long-term multilineage engraftment. Hematopoietic recovery became uncertain when reducing the CD34+ cell input (0.4 x 10(6)/kg CD34+ cells; n=3). However, platelet recovery was accelerated in all surviving cografted animals, when compared with baboons transplanted with unirradiated, unmanipulated CD34+ cells (0.5-1 x 10(6)/kg, n=4). Baboons grafted with MSCs alone (n=3) did not recover. In all cases, the nonhematopoietic toxicity remained huge. This baboon study suggests that ACT feasibility is limited. PMID- 15821762 TI - The implication of follicular lymphoma patients receiving allogeneic stem cell transplantation from donors carrying t(14;18)-positive cells. AB - We performed real-time quantitative polymerase chain reaction (RQ-PCR) in peripheral blood (PB) and/or bone marrow (BM) samples collected pre- and post transplant from 23 recipient-donor pairs receiving allogeneic stem cell transplantation (allo-SCT) for follicular lymphoma (FL). Of 23 donors, 11 had a PB and/or BM sample positive for t(14;18) (BCL2/IGH fusion) at low levels (55 kg. PMID- 15821772 TI - Neuropsychological resiliency after treatment for advanced stage neuroblastoma. AB - The purpose of this study was to describe the neuropsychological functioning of survivors of advanced stage neuroblastoma. In all, 16 survivors, diagnosed at a median of 2.8 years, who had received intensive chemotherapy and surgical treatments, were identified; 11 had received myeloablative consolidation therapy, eight with total body irradiation (TBI). All patients were evaluated with a neuropsychological assessment battery at a median age of 8.8 years. Analyses included comparison of the performances of the TBI group vs the no-TBI group; determination of whether the proportion of individuals with impaired or superior performance on each measure exceeded normative expectations; and performance indexes reflecting patterns of performance. Results indicate no significant deleterious impact of TBI and/or presence or absence of myeloablative therapy on neurocognitive and neurobehavioral functioning. For this cohort, resilience to neuropsychological vulnerability was observed, which included the emergence of a profile of full-scale IQ, verbal IQ, and mathematical achievement well above average expectations. We concluded that the results document a lack of neuropsychological morbidity among this cohort of survivors of advanced stage neuroblastoma, regardless of the inclusion of TBI. Moreover, a striking pattern of excellent neurocognitive functioning with intact neurobehavioral functioning was observed. PMID- 15821773 TI - Cyclosporin A and mini short-term methotrexate vs cyclosporin A as graft-versus host disease prophylaxis in patients with beta thalassemia major undergoing allogeneic blood and marrow transplantation. AB - We compared the effects of cyclosporin A (CsA) alone as graft-versus-host disease (GVHD) prophylaxis vs cyclosporine with short-course methotrexate (MTX) in patients with thalassemia. In all, 140 patients were enrolled in this study. The first group, of 50 patients, received CsA alone at 3 mg/kg i.v. from day -2 to +5 followed by 12.5 mg/kg p.o., which was tapered according to the patient's condition. The other group, of 90 patients, received the combination of CsA+MTX in which CsA was used with the above-mentioned dose and MTX was on 10 mg/m(2) day +1 and 6 mg/m(2) on days +3 and +6. Incidence of acute GVHD grade II-IV in the CsA group was 78% and in the CsA+MTX group was 52.2%, which was statistically significant (P=<0.001). There were no significant differences in the incidence of chronic GVHD between the two groups. The mean neutrophil engraftment to 0.5 x 10(9)/l was 14 and 23 days for CsA group and CsA+MTX group, respectively (P=<0.001). There were no significant differences for platelet recovery between the two groups. Graft failure in the CsA and CsA+MTX groups was seven (14%) and nine (10%) patients, respectively (P=0.58). Overall survival in the CsA and CsA+MTX groups was 77 and 85%, respectively. Disease-free survival in the CsA and CsA+MTX groups were 58 and 80%, respectively. PMID- 15821774 TI - Biological purging of breast cancer cell lines using a replication-competent oncolytic virus in human stem cell autografts. AB - Autologous hematological stem cell transplantation (ASCT) is used for the treatment of many hematological and several solid cancers. ASCT, however, has proven disappointing as a therapeutic strategy for breast cancer. Our group and others have previously shown that breast cancer micrometastases found in patients' apheresis products (APs) predict shorter progression-free and overall survival. The implications of this finding are twofold: (i) contaminating tumor cells (CTCs) in AP reflect a higher systemic disease burden and/or (ii) reinfused CTCs contribute to relapse/progressive disease. To date, purging strategies have been disappointing. We have previously demonstrated the oncolytic properties of reovirus in in vitro, in vivo and ex vivo systems. In the present study, we tested the hypothesis that reovirus purges CTCs in a breast cancer cell line purging model. Reovirus-infected human breast cancer cell lines (HTB 133, HTB 132, SKBR3 and MCF7) exhibited cell death within days. Admixtures of AP with cells from breast tumor cell lines, which were then exposed to reovirus, showed complete purging of CTCs (assessed via flow cytometry/tumor cell outgrowth analysis) without deleterious effect on CD34+ cells. Our results provide preclinical support for the ex vivo use of reovirus as a purging modality for breast cancer during ASCT. PMID- 15821775 TI - The role of glutathione in mammalian gametes. AB - The paper reviews a recent research on the role of glutathione (GSH) in the male and female germ cells as well as during the early stages of embryo development in mammals. In both the male and female gametes, GSH is involved in the protection of these cells against oxidative damage. Glutathione has been implicated in maintaining the meiotic spindle morphology of the oocyte. After fertilization, this thiol plays an active role in the formation of the male pronucleus, and has a beneficial effect on early embryogenesis to the blastocyst stage. GSH concentrations change within the oocytes during meiotic maturation and its synthesis is regulated by gonadotropins. Furthermore, GSH concentrations in the maturing spermatozoa gradually decline during spermatogenesis. This review also addresses the important role of cumulus cells in glutathione synthesis. PMID- 15821776 TI - A comparison of adrenal gland function in lactating dairy cows with or without ovarian follicular cysts. AB - Two experiments were conducted to determine if adrenal secretion of steroids differed between cows that formed ovarian follicular cysts and normal cycling cows. In experiment 1, lactating Jersey and Holstein cows were diagnosed as having ovarian follicular cysts (follicle diameter >or=20 mm) by rectal palpation. Following diagnosis, ovaries were examined by transrectal ultrasonography three times weekly to detect subsequent ovulation (n=8) or new cyst formation (n=9). Venous blood samples were collected daily to quantify circulating concentrations of cortisol and progesterone. The average concentration of cortisol during the 10-day period prior to ovulation was not different from the concentration prior to the formation of a new cyst. In experiment 2, secretion of cortisol and progesterone was examined in cows with ovarian follicular cysts (n=4) and cyclic, control cows in the follicular phase of the estrous cycle (n=4). An adrenocorticotropic hormone (ACTH) challenge was administered to cystic cows 4-7 days after new cyst formation and to cyclic cows in the follicular phase of the cycle (36 h after induction of luteolysis). Jugular venous blood samples were collected at -60, -30, 0, +10, +20, +30, +60, +90, +120, +180, +240, +300 and +360 minutes relative to ACTH administration. A rapid increase in both cortisol and progesterone was observed immediately following administration of ACTH in each treatment group. Peak concentrations of both steroids were achieved within 60 minutes after administration of ACTH. Concentrations of cortisol and progesterone did not differ between cystic and cyclic cows. In summary, no differences in adrenal function were detected between normal cycling cows and cows with ovarian follicular cysts. PMID- 15821777 TI - Does prolactin influence the hypothalamo-pituitary GnRH-LH system in preovulatory phase ewes? AB - The effects of prolonged infusions of prolactin (PRL) into the third ventricle of the brain of cycling ewes on the secretory activity of hypothalamic GnRH neurons and pituitary LH cells in the pars distalis during the proestrous day were studied. Mature Blackhead ewes were infused with vehicle (control, n=5) or with prolactin (200 mug/day, n=5) during 4 consecutive days prior to the next spontaneous ovulation. The dose of PRL was infused each day in 4 series of 50 mug/100 mul/h at 30-min. intervals, from 8.30 to 14.00 h. The animals were slaughtered on the 16th (proestrous) day of the estrous cycle immediately after the last infusion and their brains were fixed in situ. Plasma samples were collected for 6 h at 10 min. intervals, on days 12 (before the infusions) and 16 of the cycle. The distribution pattern, number and morphology of GnRH neurons in vehicle- and PRL-infused ewes were found to be similar and typical for the proestrous phase of the cycle. The immunoreactive (ir) GnRH stores in the median eminence were high and similar in both groups. There were no differences between control and PRL-treated ewes in the number or features of irLH cells. The area fraction and optical density for irLH cells and mRNA LHbeta-expressing cells did not differ between control and experimental groups. Irrespective of the kind of infusion, changes in LH secretion during the estrous cycle were similar in control and PRL-infused ewes. Mean plasma LH concentrations were higher (p<0.001) on day 16 compared to day 12 of the cycle. There were no differences in plasma LH concentrations or in the parameters of pulsatile LH secretion between groups. In conclusion, repeated, several-hour-long infusions of PRL into the CNS prior to the next spontaneous ovulation in ewes has no direct effect on the secretory activity of GnRH neurons, and/or the synthesis, accumulation, or tonic release of LH from the pituitary gonadotrophs. PMID- 15821778 TI - Endoglin (cd105) and S100A13 as markers of active angiogenesis in endometriosis. AB - The aim of the present study was to evaluate the expression of the neo-angiogenic marker endoglin and its localization in tissues of normal and endometriotic patients as well as to compare it with one new angiogenic marker candidate - S100A13. Human recombinant S100A13 and endoglin 35mer synthetic peptide of the intracellular domain were used for the production of rabbit polyclonal antisera. The antisera were characterized for specificity, using immunoenzyme assay (ELISA), Western blot and immunohistochemistry. Formalin-fixed, paraffin-embedded tissue sections from normal endometrium, adenomyosis, ovarian endometriosis, eutopic endometrium from different endometriotic specimens were tested by immunohistochemistry. No endoglin specific staining was observed on the microvessels of the normal endometrium. In adenomyosis and ovarian endometriosis, the expression pattern was different - endoglin was expressed in all microvessels, with an even stronger expression in the myometrial compartment. Weak endoglin-positive staining was detected in the microvessels of eutopic endometrium specimens from different endometriosis cases. In comparison to endoglin, S100A13 exhibited a moderate expression in endometrial glands of normal endometrium, but strong expression in endometriotic specimens. No S100A13 extensive staining of the microvessels was observed in normal endometrium, while in endometriosis, it exhibited very intense staining in microvascular endothelia and less intense in the perivascular area of middle to large-sized vessels. This study for the first time shows over-expression of S100A13 in endometriosis. These data show that the expression of endoglin and S100A13 corresponds to the activation of the endothelial cells in the process of endometriotic angiogenesis, suggesting a beneficial role for these two molecules as markers for actively progressing endometriotic process. PMID- 15821779 TI - Denervation of the porcine ovaries performed during the early luteal phase influenced morphology and function of the gonad. AB - We studied both morphology and steroidogenic activity of ovaries in gilts after bilateral surgical denervation performed on day 3 of the estrous cycle. Blood samples were collected from day 4 of the first estrous cycle to day 11 of the subsequent cycle. Denervation resulted in a dramatic reduction in the number or in the disappearance of tyrosine hydroxylase/dopamine-beta-hydroxylase- and/or neuropeptide Y-immunoreactive nerve fibres. On day 11 of the second cycle, the number of follicles (3-6 mm in diameter) was lower (p<0.001) in the denervated ovaries, while corpora lutea were not present. Neurectomy also led to a decrease in the concentrations of progesterone, androstendione and 17beta-estradiol in the follicular fluid originated from small (1-3 mm in diameter) as well as medium sized follicles (3-6 mm in diameter). Similar to follicular fluid, concentration of androstendione in the follicular wall of medium-sized follicles decreased in experimental gilts in comparison to that of control animals. In addition, plasma concentrations of LH and steroid hormones were lower in the control than in the experimental group. Our results show that denervation of ovaries during the early luteal phase of the estrous cycle in gilts resulted in the changes in both morphology and steroidogenic activity. These results confirm the important role of the peripheral nerves in the function of ovaries. PMID- 15821780 TI - Growth factors effects on preimplantation development of mouse embryos exposed to tumor necrosis factor alpha. AB - The success rates of assisted reproduction techniques are still unsatisfactory. Relatively few in vitro cultured embryos reach the blastocyst stage. The purpose of the study was to evaluate the protective potential of epidermal growth factor (EGF), insulin-like growth factors 1 and 2 (IGF-I, IGF-II) and stem cell factor (SCF) on in vitro development of pre-implantation mouse embryos exposed to tumor necrosis factor alpha (TNFalpha). C3B6F1 female mice were superovulated with 5 IU of pregnant mare serum gonadotropin (PMSG) and 48 h later with 5IU of equine chorionic gonadotropin (eCG). Following the second injection females were mated with DBA males. Two cell embryos were flushed out from the fallopian tubes 40 h after eCG administration. After retrieval, the embryos were divided into control and experimental media and incubated in groups of ten for 96 h (37 degrees C, 5%CO(2), in droplets of 50 microl under mineral oil). In the first part of experiment, the embryo development was tested in media containing EGF, IGF-I, IGF II, SCF, TNF-alpha (1 to 1000 ng/ml). In the second part of the study, the development of embryos was examined in medium containing 100 ng/ml TNFalpha and one of following factors: IGF-I, IGF-II; EGF or SCF (100 ng/ml). During the culture embryos were examined at 24 hours intervals to assess the embryo development. Blastocyst rate was determined following 96 hours of culture. Evaluation of total blastocyst cell number (TB) and inner cell mass (ICM) was also performed. TNFalpha significantly reduced (p<0.05) the blastocyst rates as well as TB and ICM. The examined growth factors improved the development of embryos exposed to TNFalpha. Thus, in this study, the protective action of IGF-I and II, EGF and SCF against the detrimental influence of TNFalpha was demonstrated. PMID- 15821781 TI - Daptomycin: a new drug class for the treatment of Gram-positive infections. AB - Daptomycin is the first member of a new class of bactericidal antibiotics, the cyclic lipopeptides. In September 2003, daptomycin was approved for the treatment of Gram-positive infections associated with complicated skin and skin structure infections. A key feature of daptomycin is its rapid, concentration-dependent bactericidal activity against significant Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus, glycopeptide-intermediate and resistant S. aureus and vancomycin-resistant Enterococcus faecalis and Enterococcus faecium (VRE). Daptomycin also has a unique mechanism of action, no cross-resistance with any other class of antibiotic and a relatively prolonged concentration-dependent postantibiotic effect in vitro. In the United States, daptomycin has been approved for use at a dose of 4 mg/ kg once daily in the treatment of S. aureus (including methicillin-resistant strains), three beta hemolytic streptococci (S. pyogenes, S. agalactiae and S. dysgalactiae subsp. equisimilis) and E. faecalis associated with complicated skin and skin structure infections. In addition, daptomycin is undergoing a phase III evaluation for the treatment of bacteremia and endocarditis due to S. aureus. With its once-daily dosing, favorable safety profile and low potential for resistance, daptomycin is a powerful new antibiotic therapy against Gram-positive infections. PMID- 15821782 TI - Second-generation azole antifungal agents. AB - The development of the triazole antifungal agents in the 1980s and 1990s greatly enhanced physicians' ability to treat fungal infections due to the lower toxicity of these agents compared with previous antifungal therapies. Despite the addition of these agents, there continues to be limited therapeutic choices for a number of mycoses that cause significant disease in humans. Three new agents (voriconazole, posaconazole and ravuconazole) have been developed which appear to have expanded antifungal activity compared with prior azoles. This review discusses the pharmacology, in vitro and in vivo activity, clinical studies and toxicities of these second-generation azoles. Presently, only voriconazole is available clinically and is indicated for the treatment of esophageal candidiasis, invasive aspergillosis, and refractory infections with Scedosporium apiospermum and Fusarium spp. Posaconazole and ravuconazole are still in development for human use. PMID- 15821783 TI - Cetuximab: an epidermal growth factor receptor chemeric human-murine monoclonal antibody. AB - The epidermal growth factor receptor (EGFR) is a member of the ErbB family of receptors. It is composed of extracellular domains, including a ligand-binding domain, a hydrophobic transmembrane region and a tyrosine kinase-containing cytoplasmic region. Stimulation of the EGFR by endogenous ligands, EGF or transforming growth factor-alpha (TGF-alpha), results in a conformational change in the receptor, permitting it to enter into dimers and other oligomers. Dimerization results in activation of intracellular tyrosine kinase, protein phosphorylation and stimulation of various cell signaling pathways that mediate gene transcription and cell cycle progression. The EGFR is expressed on normal human cells but higher levels of expression of the receptor have also been shown to be correlated with malignancy in a variety of cancers. In addition, expression of the EGFR by malignant cells is associated with poor prognosis and resistance to therapy. Cetuximab is a chimeric human-murine monoclonal antibody that binds competitively and with high affinity to the EGFR. Binding of the antibody to the EGFR prevents stimulation of the receptor by endogenous ligands and results in inhibition of cell proliferation, enhanced apoptosis, and reduced angiogenesis, invasiveness and metastasis. Binding of cetuximab to the receptor also results in internalization of the antibody-receptor complex which leads to an overall downregulation of EGFR expression. The EGFR is a prime target for new anticancer therapy, and other agents in development include small molecular tyrosine kinase inhibitors and antisense therapies. Preclinical studies have demonstrated that cetuximab reduces chemotherapy and radiotherapy resistance in human tumor cell lines in vitro and in nude mice bearing xenografts of human tumors. In clinical and preclinical studies cetuximab has been shown to induce response to treatment when used in combination with chemotherapy in patients previously refractory to chemotherapy. Based on these studies, cetuximab can be added to regimens using docetaxel, cisplatin, carboplatin, irinotecan, paclitaxel and fluorouracil and may add to treatment efficacy. Phase I dose-finding studies showed that saturation of cetuximab clearance occurred after administration of 400 mg/m2 as a loading dose followed by weekly infusions of 250 mg/m2. The most commonly reported adverse event associated with cetuximab treatment is an acneiform rash that occurs in 70-80% of patients treated with cetuximab. The rash is rarely dose or treatment-limiting, and may diminish in intensity with continued exposure to cetuximab. Improvement may be seen after treatment with topical antibiotic preparations, topical steroids or topical retinoids. The rash resolves fully after discontinuation of cetuximab treatment. EGFR is widely expressed in skin and skin biopsies in areas involved with the characteristic cetuximab eruption demonstrate neutrophilic folliculitis. In fact, analysis of four phase II clinical trials of cetuximab in combination with chemotherapy in patients with colorectal cancer, squamous cell carcinoma of the head and neck, or pancreatic cancer showed that development of the acneiform rash was significantly correlated with response to treatment; grade 3 rash may be especially predictive of response. It is possible that development of acneiform rash may become an important clinical prognostic marker. Serious cetuximab-related toxicities include hypersensitivity, infusion reactions and interstitial lung disease. Results of a large phase II study have shown response when used in combination with irinotecan in 22.9% of patients with EGFR-expressing, irinotecan-refractory, colorectal cancer. Cetuximab has recently been approved for this indication in the United States, Switzerland, Iceland, Norway and the 25 member states of the European Union. Other phase II and III studies show significant response to treatment in variable proportions of patients with squamous cell carcinoma of the head and neck, non-small cell lung cancer and pancreatic cancer when cetuximab is used first or second line in combination with chemotherapy. Thus, cetuximab is emerging as a very promising new therapy to be used in conjunction with existing therapies for the treatment of a spectrum of solid tumors. PMID- 15821784 TI - Renal and humoral pathophysiological actions of angiotensin II in congestive heart failure. AB - The renin-angiotensin-aldosterone axis is an integral component linking the renal humoral system to the cardiovascular system. It is involved in the normal control of blood pressure and intravascular volume. Its activity is also enhanced in pathologic states, namely congestive heart failure, in which stimulation of the axis leads to further deleterious effects on the heart. The well-established dogma that the renin-angiotensin-aldosterone system (RAAS) is a linear cascade is evolving into a vision of this system as a more complex process. It is now known that angiotensin has several subtypes. Each subtype is a ligand at several receptor subtypes and these interactions are not mutually exclusive. The aim of this review is to discuss the different angiotensin subtypes, their receptor interactions and their pathophysiological roles in humoral and renal functions in congestive heart failure. In addition, we will also review the different therapeutic approaches that interrupt the RAAS and the evidence that supports their utility in congestive heart failure (CHF). PMID- 15821785 TI - Hypothesis for the pathogenesis of type 1A diabetes. AB - Type 1A (immune-mediated) diabetes results from T-cell-mediated specific destruction of the islet beta cells that produce insulin (1). One can divide the development of diabetes into a series of overlapping stages beginning with genetic susceptibility and ending with complete beta cell destruction (2). The dominant genetic susceptibility locus is the major histocompatibility complex (MHC), also called the human leukocyte antigen (HLA) region in humans, and in particular, immune response genes DR and DQ (3). Additionally, the existence of a series of rare but informative "single-gene" disorders associated with autoimmune diabetes indicates that a number of different immunologic lesions can lead to autoimmune diabetes. The most common forms of type 1A diabetes are polygenic (multifactorial) disorders with unidentified environmental factors contributing to the disease. Given current information, it is now possible to predict the development of type 1A diabetes (4, 5), and major efforts are under way to create preventive therapies. PMID- 15821786 TI - [Lymphatic filariasis in Belem, Para State, North of Brazil and the perspective of elimination]. AB - The objective was to characterize the epidemiological situation of lymphatic filariasis in Belem, state of Para. Hemoscopic data was analyzed from 1951 through 2003. Information for the period from 1951 to 1994 was collected from reports available from the National Health Foundation. Data from 1995 to 2003 was obtained through surveys carried out in 62 city sectors, within the eight administrative districts of the city. An appreciable drop in the microfilaraemic rates was observed over the years. The percentages of parasitized individuals in the decades of 1950, 1960, 1970, 1980 and 1990, were respectively: 8.2%, 2.6%, 0.7%, 0.16% and 0.02%. In 2001, a single microfilaraemic case was diagnosed, interrupting a series of two years without registering positive cases in the city. In 2002 and 2003, hemoscopic and entomological surveys were performed simultaneously revealing no microfilariae positive individuals, nor infected mosquitoes. To maintain this trend, surveillance measures must be conducted in order to detect and promptly treat patients, to prevent the risk of resurgence of a focus apparently now controlled. PMID- 15821788 TI - Response to chemotherapy with benznidazole of clones isolated from the 21SF strain of Trypanosoma cruzi (biodeme Type II, Trypanosoma cruzi II). AB - Susceptibility to chemotherapy with benznidazole was investigated of 5 clones isolated from the 21 SF strain (biodeme Type II, Trypanosoma cruzi II). Swiss mice were infected with the parental strain for each clone and submitted to chemotherapy with benznidazole (100mg/kg/day during 90 days). Treatment determined negativity of the parasitemia. Cure rates were evaluated by parasitological cure tests. Serology was evaluated for treated animals (titers from negative to 1:640) and untreated controls (1:160 to 1:640). Cure rates varied from 30 to 100% for the 5 clones, and were 25% for the parental strain. Results suggested that the variability of response to treatment of the clonal populations of Trypanosoma cruzi II strains is responsible for the high variation in the response to chemotherapy with benznidazole and nifurtimox by strains of this biodeme. PMID- 15821787 TI - [Immunoglobulin isotype and IgG subclass profiles in American tegumentary leishmaniasis]. AB - The present work investigated the serum antibody profiles in 37 patients with American tegumentary leishmaniasis, who were attended at Hospital de Clinicas - Universidade Federal de Uberlandia, MG, Brazil. The immunoglobulin class and IgG subclass profiles were analyzed by indirect ELISA using Leishmania (Leishmania) amazonensis soluble antigen. The antibody avidity was determined by 6 M urea treatment after incubation with immunoenzymatic conjugate. It was observed that 97% of the serum samples presented anti-Leishmania antibodies for IgE class, 94.6% IgG, 57.5% IgA and 21.5% IgM class. For IgG subclasses the profiles were in the following order of frequency: IgG1>IgG3>IgG2>IgG4. High avidity of anti Leishmania IgE antibodies was found in 44.4% of the samples. On the other hand, moderate avidity of specific IgG and IgA was observed in 62.8% and 47.8% of samples, respectively. These results indicate a very complex antibody response profile against American tegumentary leishmaniasis. PMID- 15821789 TI - [Visceral leishmaniasis: a study on phlebotomine sand flies and canine infection in Montes Claros, State of Minas Gerais]. AB - Visceral leishmaniasis in Brazil was initially associated with rural areas. However, due to several environmental modifications such as deforestation, urbanization and intense migratory processes, there has been an expansion of endemic areas, leading to urbanization of the disease, mainly in the central and northeastern regions of Brazil. In the municipality of Montes Claros, located in the north of the state of Minas Gerais, an epidemiological survey on VL was carried out. A canine serological inquiry was carried out in 2002 and an entomological survey, using luminous CDC traps, was performed from September 2002 to August 2003. Canine VL prevalence showed an average infection rate of approximately 5%. An estimated 16 species comprised the phlebotomine sand fly fauna, based on a total of 1043 specimens. The predominant species was Lutzomyia longipalpis with a rate of 74%, suggesting its participation in the transmission of VL in the municipality of Montes Claros. PMID- 15821790 TI - Seroprevalence of hepatitis B virus infection and seroconvertion to anti-HBsAg in laboratory staff in Goiania, Goias. AB - Were analyzed 648 serum samples from laboratory staff in Goiania, Goias aiming detection of three serological markers of HBV: HBsAg, anti-HBsAg and anti-HBcAg. The HBsAg and anti-HBcAg positive samples were also analyzed for HBeAg, anti HBeAg and anti-HBcAgIgM markers. HBV infection rate of 24.1% was observed and, from them, 0.7% were positive for HBsAg. Viral DNA was detected by PCR in two HBsAg positive samples. A vaccination index of 74.5% and a global index of 89.9% of serological response to vaccination were observed. The direct work with biological fluids as well as cleaning workers represented significant risks for acquisition of HBV infection. The data from the present study showed an increase of the vaccination index among laboratory staff but the rates of HBV infection did not change through the years in the region. PMID- 15821791 TI - [Quantitative study of metal present in the hemolymph of Biomphalaria glabrata (Gastropoda), infected and uninfected with Schistosoma mansoni]. AB - We conducted a preliminary study to quantify and compare two concentrations of the same metals present in the hemolymph of snail Biomphalaria glabrata. In this context, we used Induction Coupled Plasma Optical Emission Spectroscopy technique (ICP-OES), to analyze the metals in the two samples (snails infected and not infected with Schistosoma mansoni). The metals studied were: aluminum, calcium, cadmium, cobalt, chromium, copper, iron, potassium, magnesium, manganese, lead and zinc. Preliminary results showed that such metals are not involved in the defense of these organisms against the parasite, when present in their tissues. PMID- 15821792 TI - [Prevalence of canine infection from endemic areas of American cutaneous leishmaniasis in Paracambi District, Rio de Janeiro State, between 1992 and 1993]. AB - In the district of Paracambi, State of Rio de Janeiro an epidemiological survey for American tegumentary leishmaniasis in canine population was carried out in endemic localities. A total of 179 dogs was registered and 138 (77.1%) examined for their clinical aspects, development of delayed hypersensitivity (DHS) with Imunoleish(R) antigen and serological responses by indirect immunofluorescent reaction and enzyme-linked immunosorbent assay. In 9 (6.5%) dogs with active cutaneous lesions or suspect scars, 66.7% were caused by Leishmania sp; 44.4% produced infection in hamsters and showed growth in culture media, which was considered to be compatible with the species of Leishmania braziliensis complex. The molecular characterization (isoenzyme and KDNA restriction profiles) defined two strains with similar profiles for L. (Viannia) braziliensis. The prevalence of canine infection estimated by the cutaneous test, IFR and ELISA was 10.1%, 16.7% and 27.8%, respectively. The presence of clinical / sub-clinical form of ATL in canine population associated with human infections suggested that the dog can act as source of infection as well as for dissemination of the disease. PMID- 15821793 TI - [Correlation between dose/plasma concentration and assessment of hepatic and renal changes in Wistar rats treated with the ROM scheme]. AB - Leprosy, a chronic granulomatous infectocontagious disease transmitted by Mycobacterium leprae, continues to be prevalent today, especially in underdeveloped countries and its paucibacillary form with a single lesion is being treated with rifampicin (600mg), ofloxacin (400mg) and minocycline (100mg) administered as a single dose (ROM scheme). Thus, the objective of the present study was to investigate the dose/plasma concentration correlation versus biochemical changes occurring in male Wistar rats receiving a single dose of rifampicin, ofloxacin and minocycline in mono- and polytherapy. Rifampicin and ofloxacin showed an increased concentration in plasma when administered in polytherapy, whereas minocycline was reduced, probably due to interference with its biotransformation and excretion. Biochemical analyses showed that rifampicin is probably responsible for hepatic and renal changes and that the medicamentous interactions involving the drug require individualized studies, especially when the drug is associated with ofloxacin and minocycline therapy. PMID- 15821794 TI - Serological screening and toxoplasmosis exposure factors among pregnant women in South of Brazil. AB - Serological screening and evaluation of exposure factors for Toxoplasma gondii transmission were conducted in 2126 pregnant women from southern Brazil. Specific antibodies against Toxoplasma gondii were presented by 74.5% (n=1583) of the pregnant women evaluated. Contact with soil was found to be the major factor for infection. PMID- 15821795 TI - [A comparative study of the parasitological techniques: Kato-Katz and coprotest]. AB - Parasitological diagnosis should be appropriately conducted with high sensitivity and specificity so that intestinal parasite can be detected as the specific treatment depends on this result. A comparative study was developed to evaluate the concordance between the Kato-Katz and coprotest methods in the detection of helminthes in 332 individuals in Pedro de Toledo. A significant difference was found in the case of the Trichuris trichiura - 16.2% with the Kato-Katz method and 7.5% with the coprotest method. Therefore, the positive and negative samples of the coprotest method were compared with the number of eggs per gram of feces (epg) obtained by the Kato-Katz method. When the coprotest method was negative, 65 epg of Trichuris trichiura were found by the Kato-Katz method and when the coprotest was positive the epg according to the Kato-Katz were far higher, 199 epg, which demonstrated that the coprotest was inferior to the Kato-Katz method regarding low intensity infections. PMID- 15821796 TI - Lues maligna in an HIV-infected patient. AB - We report such a case of malignant syphilis in a 42-year-old HIV-infected man, co infected with hepatitis B virus, who presented neurolues and the classical skin lesions of lues maligna. The serum VDRL titer, which was 1:64 at presentation, increased to 1:2,048 three months after successful therapy with penicillin, decreasing 15 months later to 1:8. PMID- 15821797 TI - Human pseudomyiasis caused by Eristalis tenax (Linnaeus) (Diptera: Syrphidae) in Goias. AB - The objective of the present work was to register the first proven cases of human pseudomyiasis due to Eristalis tenax in Goias State, Brazil, underscoring their clinical manifestations and direct relationship with hygiene. The taxonomic identification of the instars was done according to the descriptions and keys presented by James (1947), Hartley (1961) and Guimaraes & Papavero (1999). Two cases were observed. In both cases there was no evidence of apparent mental disturbance. The clinical picture of these cases was self limited. The water supply, sewer system, socioeconomic level and habits of the suspect species of the flies are criteria that should be investigated. PMID- 15821798 TI - [Entomophthoramycosis (zygomycosis) caused by Conidiobolus coronatus in Mato Grosso (Brazil): case report]. AB - Zygomycosis is a subcutaneous mycosis caused by soil fungi, such as Conidiobolus coronatus. In general, the main clinical manifestation is a chronic rhinofacial tumor. We report the first case of zygomycosis (entomophthoramycosis) caused by Conidiobolus coronatus, occurring in Mato Grosso, West Brazil. PMID- 15821799 TI - [Paracoccidioidomycosis epidemiological survey using gp43, in two cities of northwestern region of Parana, Brazil]. AB - An epidemiological survey was performed in 118 workers from northwestern Parana state using gp43 as antigen, the positivity was 43%. This result, plus the weather conditions, which are favorable for fungus development, underscores that this region is an endemic area for paracoccidioidomycosis and is a reservoir of Paracoccidioides brasiliensis. PMID- 15821800 TI - Evaluation of an immunochromatography test for malaria diagnosis under different storage conditions. AB - This study aimed to evaluate the second-generation OptiMal test for malaria diagnosis under various storage conditions. It detected all the positive samples, except for two Plasmodium malariae samples. Further research evaluating diverse environmental conditions are important for ICT test applicability in Brazilian malaria areas. PMID- 15821801 TI - Comparison of the thick smear and Kato-Katz techniques for diagnosis of intestinal helminth infections. AB - This study compared the efficiency of Kato-Katz thick smear and thick smear techniques for the diagnosis of intestinal helminths. The sensitivity of the thick smear technique was higher than that of the Kato-Katz method for the diagnosis of all helminths except Schistosoma mansoni. PMID- 15821802 TI - [Contamination of public gardens along seafront of Praia Grande City, Sao Paulo, Brazil, by eggs of Ancylostoma and Toxocara in dogs feces]. AB - Contamination of the lawns and flower beds along the seafront of Praia Grande, SP, Brazil, by eggs of Ancylostoma and Toxocara in fecal samples of dogs was evaluated. Of the total samples analyzed, 45.9% were contaminated with Ancylostoma eggs and 1.2% with Toxocara eggs. PMID- 15821803 TI - [Entomological survey and infectivity during malaria outbreak in the Anajas municipality, Para State]. AB - An entomological survey was conducted in the urban area of Anajas municipality, Para State, in order to verify the risk of malaria transmission. Although the density of Anopheles species was low, the anopheline infectivity rate of 6% demonstrated there is a high risk of infection by the malaria parasites. PMID- 15821804 TI - In vitro maintenance of Angiostrongylus costaricensis does not provide physiological conditions for egg laying. AB - Fifteen pairs (male/female) of Angiostrongylus costaricensis were kept in vitro in Waymouth medium for three days to evaluate the amount and duration of egg laying. At 24, 48 and 72 hours, the mean egg counts were 321, 24 and 4 eggs/10 microliters, respectively. Most of the eggs were eliminated within the first 24 hours, suggesting they are expelled under non-physiological conditions. These results indicate that in vitro conditions are not appropriate for drug trials of egg-laying inhibitors for treatment of abdominal angiostrongylosis. PMID- 15821805 TI - Cryptococcal meningoencephalitis and pulmonary nodule in a non-HIV-infected immunocompetent patient. PMID- 15821806 TI - [Mario Rubens Guimaraes Montenegro]. PMID- 15821807 TI - Stroke in Brazil: a neglected disease. PMID- 15821808 TI - Continuing medical education in Brazil: what about obstetricians and gynecologists? AB - CONTEXT: In Brazil, obstetricians and gynecologists are not required to submit to periodical evaluations to ascertain their professional competence in dealing with new concepts and therapies. OBJECTIVES: To evaluate the performance of a group of obstetricians and gynecologists on a written evidence-based obstetrics test and determine their opinions and use of systematic reviews. TYPE OF STUDY: Prospective cohort. SETTING: Brazilian Obstetrics and Gynecology Congress 2001. METHODS: 230 doctors agreed to participate in the study during a national obstetrics and gynecology congress. Participants took an individual anonymous written multiple-choice test with seven questions on clinical obstetrics, one question on the interpretation of a meta-analysis graph and two questions on their opinions and actual use of systematic reviews. Scores were analyzed and compared after grouping the participants according to year of graduation, residence training, doctoral program and faculty status. RESULTS: The general average score was 49.2 +/- 17.4. The scores tended to decline as the years since graduation advanced. Doctors who graduated in the last five years had higher scores than those who graduated over 25 years ago (52.2 versus 42.9). The performance did not vary according to medical residence, postgraduate program or teaching status. While 98.2% considered systematic reviews relevant, only 54.9% said that they routinely used this source of information. DISCUSSION: The participants' average score was low, even though they were highly qualified and trained. Despite the limitations of the study, the results are worrisome. If motivated physicians participating in a national congress obtained such low scores, we can speculate that the results might be even worse among other doctors that do not attend these events. CONCLUSIONS: These findings suggest that Brazilian obstetricians and gynecologists could benefit from continuing medical education and raise questions about the recycling methods currently available. PMID- 15821809 TI - The impact of psychosocial factors on the risk of erectile dysfunction and inhibition of sexual desire in a sample of the Brazilian population. AB - CONTEXT: Sexual dysfunctions can have origins in physical, psychological and psychosocial factors. OBJECTIVE: To describe the frequency of erectile dysfunction (ED) and female inhibition of sexual desire (ISD) in a Brazilian sample, and to estimate the risks of these dysfunctions. TYPE OF STUDY: Non random survey. SETTING: Ten Brazilian cities. METHODS: 2,835 subjects (53% women) aged over 18 years answered a questionnaire about their general health and sex life. The chi-squared test and multivariate logistic regression were used. Values of p < or = 0.05 were considered significant. RESULTS: The women's average age was 36.6 years (+/- 13.3) and the men's was 39.5 (+/- 13.3). 14.7% of men presented moderate/complete ED and 34.6% of women presented ISD. Depression was mentioned by 16.8% of men and 29.7% of women. The chances of having ED and ISD were higher for subjects who had had lower school attainment. Lack of a job and depression gave rise to 1.5 times (95% CI: 1.0 - 2.3) and 1.9 times (95% CI: 1.2 3.0) greater chances of ED respectively. Compared with men aged up to 25 years, those aged 41-60 had 1.9 times (95% CI: 1.0 - 3.4) and those aged 61 and over had 5.4 times (95% CI: 2.3 - 12.6) greater risk of ED. For women, lack of a job gave rise to 1.5 times (95% CI: 1.1 - 1.9) greater chance of ISD; depression was not associated with higher risk. Compared with women aged up to 25 years, those aged 41-60 and 61 or over had, respectively, 2.9 times (95% CI: 2.0 - 4.1) and 7.5 times (95% CI: 3.0 -18.6) greater risk of ISD. DISCUSSION: Increasing unemployment has affected the whole population, but especially those with lower levels of school attainment. Such levels are positively linked with presence of sexual dysfunctions. CONCLUSION: Lack of jobs, age and low school attainment are risks for the sexual dysfunctions studied. Depression increased the risk of ED but not female ISD. PMID- 15821810 TI - Association of anticardiolipin antibody and C677T in methylenetetrahydrofolate reductase mutation in women with recurrent spontaneous abortions: a new path to thrombophilia? AB - CONTEXT: Recurrent spontaneous abortion (RSA) has been associated with venous thrombosis in the mother. Acquired and inherited thrombophilia factors are possible causes. OBJECTIVE: To evaluate the association between thrombogenic factors and recurrent spontaneous abortion. TYPE OF STUDY: Case-control study. SETTING: Centro de Atencao Integral a Saude da Mulher, Universidade Estadual de Campinas. METHODS: 40 ml of blood was collected from 88 women attending an RSA clinic and 88 fertile women attending a family planning clinic, to evaluate the presence of acquired and inherited thrombophilia factors. Anticardiolipin antibodies (ACA), lupus anticoagulant and deficiencies of proteins C and S and antithrombin III were evaluated by enzyme-linked immunosorbent assay (ELISA), dilute Russell Viper Venom time (dRVVT), coagulometric and chromogenic methods. DNA was amplified by the polymerase chain reaction (PCR) to study factor V Leiden and G20210A mutations in the prothrombin gene and C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene. Data were analyzed using odds ratios and a regression model for age adjustment. Fishers exact test was used to evaluate statistical relationships between associated factors and RSA. RESULTS: ACA was detected in 11 women with RSA and one fertile woman. Heterozygous C677T was detected in 59 women with RSA and 35 fertile women. Concomitant presence of ACA and C677T was found in eight women with RSA and no fertile women (p < 0.01). DISCUSSION: The meaning of the association between C677T mutation in the MTHFR gene and ACA is still not clear. It is possible that an inherited factor that alone would not strongly predispose a woman to thrombosis could, when associated with an acquired factor, start the process and increase the likelihood of thrombosis expression. CONCLUSIONS: ACA and C677T in the MTHFR gene are statistically associated with RSA. The association of these two conditions is a new finding in thrombogenic factors and RSA. PMID- 15821811 TI - Diagnosis of malignancies in children with musculoskeletal complaints. AB - CONTEXT: Musculoskeletal complaints may be associated with neoplasias as an initial manifestation of the disease. When these symptoms predominate at the onset of the disease, the differential diagnosis includes several rheumatic diseases. OBJECTIVE: To assess the frequency, clinical features and types of cancer manifested in children presenting with musculoskeletal complaints over a seven-year period. TYPE OF STUDY: Retrospective. SETTING: Discipline of Allergy, Clinical Immunology and Rheumatology, Universidade Federal de Sao Paulo-Escola Paulista de Medicina. METHODS: The medical records of patients with musculoskeletal complaints and final diagnosis of malignant disease were reviewed. The data collected were: age when symptoms initially presented, age at diagnosis, clinical features presented, laboratory findings, and the initial and final diagnoses. RESULTS: A final diagnosis of cancer was found in nine out of 3,528 patients (0.25%) whose initial symptom was musculoskeletal pain. The mean time between disease onset and final diagnosis was five months. The most common features presented were pauciarticular arthritis or arthralgia involving the large joints. Juvenile rheumatoid arthritis was the most frequent initial diagnosis, in four out of nine patients. Anemia was the most frequent initial hematological change. Six out of eight patients had an increased erythrocyte sedimentation rate. The lactate dehydrogenase level was raised in five out of eight patients. The malignancies found included acute lymphocytic leukemia, acute myeloid leukemia, lymphoma, neuroblastoma and Ewing's sarcoma. DISCUSSION: The frequency of neoplasia in patients with musculoskeletal pain resembled reports in the literature. Consumptive symptoms were not the warning signal in most of our patients. In subsidiary tests, progressive anemia was the most common finding, although the peripheral blood cell count may continue to be normal for weeks or months after symptom onset. CONCLUSION: Malignancy always needs to be ruled out in cases of children with musculoskeletal complaints. Uncharacteristic clinical manifestations and nonspecific laboratory tests may cause difficulty in the final diagnosis, and rigorous investigation should be performed. PMID- 15821812 TI - Brazilian union actions for workers' health protection. AB - CONTEXT: Many authors have emphasized the importance of worker strength through unionized organizations, in relation to the improvement of working procedures, and have reported on the decisiveness of labor movement actions in achieving modifications within the field of work and health. OBJECTIVE: To describe the ways in which Brazilian unions have tried to intervene in health-illness and work processes, identifying the existence of commonality in union actions in this field. TYPE OF STUDY: Qualitative study. SETTING: Postgraduate Program, Environmental Health Department, Faculdade de Saude Publica, Universidade de Sao Paulo, Sao Paulo, Brazil. METHODS: Union health advisers and directors were interviewed. Documents relating to union action towards protecting workers' health were collected and analyzed. RESULTS: Unions articulate actions regarding workers' health of a technical and political nature that involve many aspects and high complexity. These have been divided into thematic categories for better analysis. DISCUSSION: Union actions regarding workers' health in Brazil are restricted to some unions, located mainly in the southern, southeastern and northeastern regions of the country. Nonetheless, the unions undertaking such actions represent many professions of great economic and political importance. CONCLUSIONS: The recent changes in health and safety at work regulations, recognition of professional diseases, creation of workers' health services and programs within the unified health system, and operational improvements in companies' specialized safety and occupational medicine services, all basically result from union action. There is commonality of union action in this field in its seeking of technical and political strengthening for all workers and their general and local representation. This has the objective of benefiting collective bargaining between employers and workers. Inter-institutional action on behalf of workers' rights guarantees and amplifies the improvement of health and working conditions. PMID- 15821813 TI - Obstructive jaundice secondary to bile duct involvement with Hodgkin's disease: a case report. AB - CONTEXT: Obstructive jaundice due to lymphoma is very rare. It may be difficult to distinguish between this condition and a large number of causes of extrahepatic bile duct obstruction, even by endoscopic retrograde cholangiography. Its prognosis is poor. Combined chemotherapy and/or radiotherapy with bile duct drainage is a therapeutic option. CASE REPORT: We describe a case of obstructive jaundice as the initial presentation of Hodgkin's disease. After chemotherapy and endoscopic bile duct stenting, it was noted that the enlarged lymph nodes, jaundice and bile duct dilation disappeared. PMID- 15821814 TI - Rhabdomyolysis and acute renal failure after strenuous exercise and alcohol abuse: case report and literature review. AB - CONTEXT: Rhabdomyolysis is a severe and life-threatening condition in which skeletal muscle is damaged. Acute renal failure due to rhabdomyolysis has been widely described and its main pathophysiological mechanisms are renal vasoconstriction, intraluminal cast formation and direct myoglobin toxicity. OBJECTIVE: To report on a case of acute renal failure (ARF) induced by rhabdomyolysis due to strenuous exercise and alcohol abuse and to describe the pathophysiology of this type of ARF. CASE REPORT: A 39-year-old man arrived at the hospital emergency service with swollen legs and lower extremity compartment syndrome. He was oliguric and had serum creatinine and urea levels of 8.1 mg/dl and 195 mg/dl, respectively. The diagnosis of rhabdomyolysis was made through clinical and laboratory findings (creatine kinase activity of 26320 IU/l). The initial treatment consisted of fluid replacement and forced diuresis. The specific treatment for compartment syndrome, such as fasciotomy, was avoided in order to prevent infection. Partial recovery of renal function was recorded, after ten hemodialysis sessions. Complete recovery was observed after two months of follow-up. PMID- 15821815 TI - Vascular injuries during gynecological laparoscopy--the vascular surgeon's advice. AB - CONTEXT: Iatrogenic vascular problems due to laparoscopy are a well recognized problem and lead to significant repercussions. In this context, a ten-year review of cases topic is presented, based on experience gained while heading two important vascular surgery services. CASES: Five patients with vascular injuries during elective laparoscopy are described. These patients presented with seven lesions of iliac vessels. All cases were evaluated immediately and required laparotomy, provisional hemostasis and urgent attendance by a vascular surgeon. Direct suturing was performed in three cases. One aortoiliac bypass and one ilioiliac reversed venous graft were made. Venous lesions were sutured. One case of a point-like perforation of the small bowel was found. There were no deaths and no complications during the postoperative period. DISCUSSION: Important points on this subject are made, and advice is given. There needs to be immediate recognition of the vascular injury, and expert repair by a vascular surgeon is recommended, in order to significantly reduce the degree of complications. PMID- 15821816 TI - In response to the editorial: "Brazil is getting older: some lessons from the Bambui Health and Aging Study". PMID- 15821817 TI - Severe suicide attempts in young adults: suicide intent is correlated with medical lethality. PMID- 15821818 TI - Genetics of platelet glycoprotein receptors: risk of thrombotic events and pharmacogenetic implications. AB - Platelet aggregation and coronary thrombosis have a central role in the development of acute coronary syndromes and myocardial infarction (MI). Therapies aimed at inhibiting platelet aggregation have shown great benefit in individuals with coronary disease or with multiple risk factors for coronary disease. Genetic variation in platelet surface receptors mediating thrombus formation has been suggested to be associated with platelet hyperreactivity, with increased risk of MI and possibly with the benefit received from various antithrombotic drug treatments. This review focuses on discrepancies and their likely explanations in studies on platelet glycoprotein genetics. Current knowledge on important issues concerning coronary event phenotypes and pharmacogenetics is analyzed. Possible future applicability of these data to patient treatment is also discussed. PMID- 15821819 TI - High-dose intravenous dalteparin can be monitored effectively using standard coagulation times. AB - The objective of this study was to examine the pharmacokinetics of intravenous dalteparin (Fragmin, Pharmacia-Upjohn, Peapack, NJ) and to assess the accuracy of standard coagulation-based monitoring techniques as an estimate of drug concentration with which to guide dosing. Knowledge of the kinetic behavior of low-molecular-weight heparins (LMWHs) and the possible utility of coagulation times for monitoring may aid in the development of safe and effective dosing algorithms for percutaneous coronary interventions. Twenty normal volunteers were treated at 2-week intervals with each of three intravenous dalteparin doses. Measurement of anti-IIa, anti-Xa, activated partial thromboplastin time (aPTT), activated clotting time (ACT), and low-range ACT was performed at baseline and at seven additional time points over 8 hours. The half-life of intravenous dalteparin is 77 minutes with slight dose-related variation. The aPTT, LR-ACT, and standard ACT are prolonged after dalteparin administration with the increase closely correlated to anti-Xa activity (aPTT, r = 0.85; LR-ACT, r = 0.79). Classification of anticoagulation intensity range using aPTT or LR-ACT in comparison to anti-Xa activity (0.5-0.99, 1.0-1.49, 1.5-2, >2) displays a level of agreement (kappa: aPTT = 0.69, LR-ACT = 0.59) that is comparable to values reported for coagulation time guidance of unfractionated heparin administration. Standard coagulation times are sensitive to the anticoagulant effect of dalteparin with a degree of correlation that suggests their utility for estimating drug concentration during high dose therapy. Trials establishing a relationship between monitoring and clinical efficacy, and the risk/reward of different treatment ranges alone or in combination with GPIIb/IIIa inhibitors and clopidogrel, are necessary. PMID- 15821820 TI - Effect of antihypertensive treatment on plasma fibrinogen and serum HDL levels in patients with essential hypertension. AB - The influence of hypertension, and its treatment, on circulating lipid and fibrinogen (Fib) concentrations in patients with essential hypertension was investigated. The lipid profile and Fib levels were measured in 353 patients (131 men) with essential hypertension. Their median age was 60 years (range: 18-85 years). All patients had normal results from liver, renal, and thyroid function tests. There were 162 patients (45.9%) who were not receiving antihypertensive treatment. Of the remaining patients, 117 were taking 'lipid-hostile' beta blockers, thiazide diuretics) antihypertensives and 74 were taking 'lipid neutral' (angiotensin-converting enzyme inhibitors, calcium channel blockers, angiotensin-II receptor blockers) agents. Patients who were taking 'lipid hostile' antihypertensive drugs had significantly higher plasma Fib concentrations when compared with those taking 'lipid-neutral' antihypertensives or those not receiving antihypertensive treatment. These differences were not attributable to established factors that influence plasma Fib levels, since when smokers and patients with diabetes mellitus and/or vascular disease were excluded, the difference remained significant. In addition, in these more homogeneous groups, patients receiving 'lipid-neutral' treatment had significantly higher serum high-density lipoprotein (HDL) cholesterol levels when compared with both those taking 'lipid-hostile' antihypertensives and untreated ones. There were no significant differences in the other lipid variables, independently of the presence/absence or the type of antihypertensives. These results suggest that antihypertensive drugs have additional effects, beyond lowering blood pressure, on other vascular risk factors, like Fib and HDL. These effects may depend on the type of drug used. PMID- 15821821 TI - Endothelial damage and hemostatic markers in patients with uncomplicated mild-to moderate hypertension and relationship with risk factors. AB - Endothelial damage, high fibrinogen levels, and platelet activity are the important accelerating factors for the development of hypertension (HT). von Willebrand factor (vWF; endothelial damage marker), fibrinogen levels, and platelet aggregability were compared between patients with uncomplicated, mild-to moderate hypertension and healthy subjects. The relationship between traditional cardiovascular risk factors and endothelial damage and prothrombotic state was evaluated. One hundred sixty-nine (54 males, 115 females) patients with untreated and uncomplicated mild-to-moderate HT, and age, gender, and body mass index matched 124 (58 males, 83 females) healthy subjects were enrolled in this study. Plasma vWF, fibrinogen levels, adenosine diphosphate-induced platelet aggregability, insulin, glucose, serum lipids, and uric acid were measured. Patients with HT had significantly increased fibrinogen, vWF, platelet number and aggregability induced by adenosine diphosphate, triglycerides, total/HDL-C, glucose, uric acid levels, and insulin resistance than control group. vWF and hemostatic markers were comparable between smoker and nonsmoker subjects. Platelet aggregability was positively related to systolic and diastolic blood pressure, and vWF. Fibrinogen was positively associated with body mass index (BMI), systolic and diastolic blood pressure, total cholesterol (TC), uric acid, vWF, and insulin resistance. vWF was significantly related to age, systolic blood pressure, TC, LDL-C, and total/HDL-C. Systolic blood pressure was independently related to vWF. vWF and diastolic blood pressure were significant predictors for adenosine diphosphate-induced platelet aggregability. Systolic blood pressure and vWF were independent predictors for fibrinogen levels. Uncomplicated mild-to moderate HT had endothelial damage and is associated with a prothrombotic state. Traditional cardiovascular risk factors such as age, BMI, dyslipidemia, and insulin resistance are important contributors to the development of endothelial damage and a prothrombotic state. Therefore, it is important to control these cardiovascular risk factors along with proper treatment of HT for preventing target organ damage in mild-to-moderate HT. PMID- 15821822 TI - Prognostic impact of aortic calcification index and ankle-arm blood pressure index in patients under hemodialysis. AB - The mortality rate is high in patients receiving hemodialysis (HD), atherosclerotic diseases being the major cause of death. As marker of clinical outcome, a prospective examination of atherosclerotic tests and atherosclerotic risk factors in patients receiving HD was performed. On April 2000, 84 patients receiving HD were followed up until April 2002. At entry to the study, several atherosclerotic tests, including ankle-arm blood pressure index (API), aortic calcification index (ACI), and atherosclerotic risk factors, were performed. In 36 patients with old thrombotic events, 26 had new thrombotic events. Of 48 patients without previous thrombotic events, 15 had new thrombotic events. During 2 years, 41 patients had new thrombotic events and 15 patients died due to thrombotic disorders. The HD durations were significantly longer in non-survivors than survivors and the body mass index was lower in non-survivors than survivors. There was a significant difference in the values of ACI and API between survivors and non-survivors, and between patients with and without thrombotic events. These findings suggest that the ACI and API have a prognostic value because they might predict the occurrence of thrombosis. PMID- 15821823 TI - A protocol for the use of enoxaparin during pregnancy: results from 85 pregnancies including 13 multiple gestation pregnancies. AB - Many practitioners consider low-molecular-weight heparin (LMWH) an alternative to unfractionated heparin, although there are limited safety data regarding maternal and fetal outcomes in patients using an LMWH during pregnancy. A retrospective chart review was performed on 72 patients with thrombophilia exposed to the LMWH, enoxaparin, during pregnancy. Eighty-five pregnancies resulted in 93 of 99 potential live births. Eleven of 12 twin pregnancies and one triplet pregnancy were successful. One preterm live birth infant of 33 weeks' gestation did not survive. Three patients with thrombophilia spontaneously aborted. A patient receiving injectable fertility treatment had spontaneously aborted one twin at 5 weeks' gestation. One patient terminated the pregnancy after discovering the presence of Down's syndrome. The mean maximum dose required to achieve a therapeutic anti-Xa level of 0.2-0.4 IU/mL at 5 to 6 hours following administration, was 38.1 mg every 12 hours (median 35 mg, range 30-75 mg every 12 hours). The mean anti-Xa level was 0.28 IU/mL (median 0.3, range 0.05-0.8 IU/mL). A total of nine patients experienced bleeding events, two requiring discontinuation of enoxaparin for the remainder of the pregnancy. Two patients experienced injection site reactions requiring discontinuation of enoxaparin. Three patients developed preeclampsia, two placenta abruptio, and one placenta previa. No thromboembolic complications or osteoporotic fractures had occurred. Enoxaparin was safe and effective for preventing thromboembolism and adverse obstetrical complications in our patients, including 12 of 13 multiple gestation pregnancies. PMID- 15821824 TI - Prevalence of thromboembolic disease including superior vena cava and brachiocephalic veins. AB - Thrombosis of the brachiocephalic veins or superior vena cava (SVC) is rare. This study was conducted to determine the prevalence and characteristics of thrombosis of brachiocephalic veins and SVC , and its association with symptomatic pulmonary embolism (PE). The prevalence of thrombosis involving the brachiocephalic veins and SVC was evaluated retrospectively at a university hospital during the 3-year period. Patients were identified by hospital records and review of computer generated lists of of all venograms, contrast-enhanced chest computed tomography, and magnetic resonance angiograms of the upper extremity and SVC. Thrombosis of the brachiocephalic veins and SVC was diagnosed in 33 (0.03 %) of 100,942 patients of all ages [(32 of 70,751 adult patients >or= 20 years; 0.04%)]. Twenty three (70%) patients initially had secondary thrombosis with multiple risk factors: associated with malignancy in 14 (42%) patients, chronic disorders in 13 (39%) patients, central venous lines (CVL) and peripheral venous lines (PVL) in 9 (27%) patients, and thrombophilia in 10 (38%) of 26 patients. Swelling of the arm, head, and neck was present in 32 (97%) patients. Symptomatic PE developed before thrombosis being treated in 12 (36%) patients. All patients except eight (three, thrombolytic; five, thrombectomy) received anticoagulant therapy. Thrombosis of the SVC and brachiocephalic veins is an uncommon but serious complication in patients with malignancy, chronic disorders, CVL, PVL, and thrombophilia. Because it is important clinical problem with frequent PE, the patients with appropriate clinical findings should be diagnosed early with imaging tests and treated with anticoagulant drugs. PMID- 15821825 TI - Clinicohematologic spectrum in patients with lupus anticoagulant. AB - A retrospective analysis of clinicohematologic parameters of 25 patients with lupus anticoagulant was carried out. The hematologic tests included dilute Russel viper venom test (dRVVT), kaolin clotting time (KCT), activated partial thromboplastin time, and prothrombin time. The diagnosis of lupus anticoagulants was based on the presence of prolonged KCT/dRVVT, its absence of correction with normal plasma and correction by phospholipids. Specific factor assays and platelet aggregation studies were performed wherever required. Ten patients (40%) had thrombosis, which was venous in 5 (50%) and arterial in 4 (40%). One patient (10%) had both arterial and venous thrombosis and presented with catastrophic antiphospholipid syndrome. Eighteen female patients conceived. Four (22%) of these had recurrent first trimester abortion. Five (20%) patients had bleeding manifestations. One (4%) of these had hypoprothrombinemia and was diagnosed to have hypoprothrombinemia lupus anticoagulant syndrome. However in two of these patients, no cause of bleeding could be identified other than the presence of lupus anticoagulants. It is concluded that patients with lupus anticoagulant have a varied spectrum of hemostatic disorders. Bleeding may sometimes occur in these patients due to associated thrombocytopenia or associated factor inhibitors. Rarely, it may occur due to presence of lupus anticoagulants alone. PMID- 15821826 TI - The plasma levels of prostanoids and plasminogen activator inhibitor-1 in primary and secondary thrombocytosis. AB - An elevated platelet count is a common finding in both hospitalized and ambulatory patients. Thrombosis and bleeding complications are more frequently observed in patients with clonal thrombocytosis than secondary thrombocytosis. The aim of this study was to investigate the behaviors of plasminogen activator inhibitor type 1 (PAI-1), the inhibitor of fibrinolysis; and thromboxane A2 and 6 keto-PGF1 alpha, the products of endoperoxides, in 16 patients affected with clonal thrombocytemia as compared with 16 patients with reactive thrombocytosis and 15 normal controls. In the clonal thrombocytemia group, plasma levels of PAI 1 antigen and activity were significantly higher than both reactive thrombocytosis and control group. Plasma levels of 6-keto-PGF1alpha were significantly higher in the clonal thrombocytemia group than the other two groups and also higher in the reactive thrombocytosis group than the control group, which was also significant. This study confirms that arachidonate metabolism is frequently deranged in patients with thrombocytosis and hypofibrinolysis due to increased PAI-1 plasma levels as shown in the clonal thrombocytosis group. This may explain the thrombotic tendency in myeloproliferative disorders. PMID- 15821827 TI - The increase of blood anticardiolipin antibody depends on the underlying etiology in cerebral ischemia. AB - Although anticardiolipin antibody (aCL) has been suggested to be a potent risk factor for thrombosis and atherosclerosis in multiple arterial beds, conflicting results still exist between aCL and cerebral ischemia in the general stroke population. To elucidate if this discrepancy relates to the heterogeneity of underlying etiologies, blood beta(2)-glycoprotein I dependent-aCL was evaluated in 432 Taiwanese adults associated with cerebral ischemia who were classified into five subtypes according to their causes of cerebral ischemia. The results were compared with those in 100 healthy controls. A definite increase of aCL-IgG isotype was found in 41 patients (9.35%) and four controls (4.0%). The relative risk was 2.52. The frequency of increased aCL-IgG was 12.2%, 12.8%, 8.8%, 3.9%, and 3.5% in patients with large-artery atherosclerotic disease, stroke of unknown etiology, small-artery occlusive disease, cardioembolism, and stroke of other known etiology, respectively. Only patient with large-artery atherosclerotic disease (p<0.025) and stroke of unknown etiology (p<0.05) had a higher frequency of increased aCL than control. The frequencies of abnormal result of activated partial thromboplastin time, antinuclear factor, Coombs' test, and venereal disease research laboratory were 2.84%, 1.22%, 1.02%, and 1.34% in these 41 patients, respectively. Accordingly, aCL-IgG selectively increases in patients with large-artery atherosclerosis and stroke of unknown etiology, reflecting selective activation of humoral immunity for aCL in the pathogenesis of cerebral ischemia. PMID- 15821828 TI - The role of plasma exchange in HELLP syndrome. AB - Plasma exchange therapy has been successfully used in selected patients with hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome who have organ failure or refractory to treatment. There is no prospective study regarding plasma exchange and its effect in HELLP syndrome. The aim of this study was to investigate the effects of early postpartum use of plasma exchange in patients with HELLP syndrome on outcomes. The mortality rate and the recovery times were compared in patients with HELLP syndrome treated with plasma exchange and historic control group of patients treated conservatively. During a 3-year period (between April 2000 and December 2003), 29 consecutive patients with HELLP syndrome were treated with single or multiple plasma exchange by using fresh frozen plasma at post-partum period. The control group consist of 26 patients with HELLP syndrome treated between 1993 and 1999. Maternal mortality rate was 23.1% in the control group; there was no death in plasma exchange group; and the mortality rate was significantly higher in the control group (p=0.006). The length of stay at the intensive care unit was shorter in the plasma exchange group (p<0.0001). Rapid improvement of the platelet, aspartate aminotransferase, alanine aminotransferase, and lactic dehydrogenase levels were observed in the plasma exchange group. This study showed that postpartum early plasma exchange therapy improves treatment outcomes in patients with severe HELLP syndrome. PMID- 15821829 TI - Study on the phylogenetic tree of human platelet glycoproteins. AB - Platelet glycoprotein is an important group of glycoproteins on platelets. Several types of platelet glycoproteins have been studied for their functions in the hemostasis system. A bioinformatic analysis was performed to find out how the platelet glycoproteins' genes are related to each other. A multiple sequence alignment phylogenetic tree was performed to present the family tree of the human platelet glycoproteins recorded in the genomic database, ExPASY. These derived sequences from the database were processed by ClustalW and subsequently used for preparation of the distance matrix by Phylip protdist. The final generated phylogenetic tree of human platelet glycoproteins was presented and discussed. PMID- 15821830 TI - Beta2 glycoprotein 1 in Indian patients with SLE. AB - Forty-five patients with systemic lupus erythematosus (SLE) were investigated to evaluate the role of antiphospholipid antibodies in causation of thrombosis in Indians. The antiphospholipid antibodies studied included lupus anticoagulant (LAC), anticardiolipin antibodies (aCL), and anti-beta(2)-glycoprotein 1 (a beta(2)-GP1). Twenty-seven patients (60%) had clinical manifestations of antiphospholipid antibody syndrome. Nineteen patients (42.2%) had a history of thrombosis, and eight (17.7%) had a history of recurrent fetal loss. aBeta(2)-GP1 was (IgG) was positive in 23 (51.1%), aCL in 13 (28.8%), and LAC in four (8.8%). Of 19 patients with thrombosis, 14 (73.6%) were positive for abeta(2)-GP1, eight (42.1%) for aCL, and none of them was positive for LAC. Of the eight patients with recurrent fetal loss, two (25%) patients were positive for beta(2)-GP1, five (62.5%) for aCL, and one (12.5%) for LAC. Of 18 patients without any manifestations of antiphospholipid syndrome (APS), seven patients (38.8%) were positive for abeta(2)-GP1, and three (16.6%) for aCL and LAC each. It is concluded that presence of abeta(2)-GP1 increases the risk of thrombosis and therefore should be looked for in all cases of SLE to consider prophylactic antithrombotic therapy in these patients. PMID- 15821831 TI - Global fibrinolytic capacity increased exponentially in metastatic colorectal cancer. AB - Colorectal cancers are one of the most common malignancies associated with coagulation abnormalities ranging from asymptomatic laboratory changes to massive thromboembolism or hemorrhage. It was previously shown that global fibrinolytic was increased in non-metastatic colorectal cancer. In this study global fibrinolytic capacity was measured in patients with colorectal cancer and metastatic liver disease, which always more commonly displays various coagulation disorders. Nineteen patients with biopsy-proven colorectal cancer, 30 patients with metastatic colorectal cancer, and 20 healthy control subjects were involved into the study. Using standart silicated fibrin pellets and tissue plasminogen activator, fibrinolytic capacity of the plasmas was detected with the amount of d dimer produced before the reaction was stopped by adding aprotinin to the medium. Mean global fibrinolytic capacity (GFC) was increased to higher levels in patients with metastatic disease compared to levels in non-metastatic disease (p<0.05). Fibrinogen/GFC ratio correlated to the increase of d-dimer levels. Global fibrinolytic capacity was much higher in metastatic disease, reflecting a progression to overt disseminated intravascular coagulation. PMID- 15821832 TI - Diagnosis of antiphospholipid syndrome during the diagnostic workup of second trimester fetal bradycardia. AB - Antiphospholipid syndrome (APS) is an autoimmune disorder associated with either arterial and/or venous thrombosis (1). Two cases of APS, which were diagnosed in the second trimester during the diagnostic workup of fetal bradycardia, are presented. PMID- 15821833 TI - Hyperhomocysteinemic diabetic cardiomyopathy: oxidative stress, remodeling, and endothelial-myocyte uncoupling. AB - Accumulation of oxidized-matrix (fibrosis) between the endothelium (the endothelial cells embedded among the myocytes) and cardiomyocytes is a hallmark of diabetes mellitus and causes diastolic impairment. In diabetes mellitus, elevated levels of homocysteine activate matrix metalloproteinase and disconnect the endothelium from myocytes. Extracellular matrix functionally links the endothelium to the cardiomyocyte and is important for their synchronization. However, in diabetes mellitus, a disconnection is caused by activated metalloproteinase, with subsequent accumulation of oxidized matrix between the endothelium and myocyte. This contributes to endothelial-myocyte uncoupling and leads to impaired diastolic relaxation of the heart in diabetes mellitus. Elevated levels of homocysteine in diabetes are attributed to impaired homocysteine metabolism by glucose and insulin and decreased renal clearance. Homocysteine induces oxidative stress and is inversely related to the expression of peroxisome proliferators activated receptor (PPAR). Several lines of evidence suggest that ablation of the matrix metalloproteinase (MMP-9) gene ameliorates the endothelial-myocyte uncoupling in diabetes mellitus. Homocysteine competes for, and decreases the PPARgammaactivity. In diabetes mellitus, endothelial myocyte uncoupling is associated with matrix metalloproteinase activation and decreased PPARgamma activity. The purpose of this review is to discuss the role of endothelial-myocyte uncoupling in diabetes mellitus and increased levels of homocysteine, causing activation of latent metalloproteinases, decreased levels of thioredoxin and peroxiredoxin, and cardiac tissue inhibitor of metalloproteinase (CIMP) in response to antagonizing PPARgamma. PMID- 15821834 TI - Emerging role of direct thrombin inhibitors in nonvalvular atrial fibrillation: potential and peril. AB - Ximelagatran, an emerging new direct, oral, fixed-dose thrombin inhibitor may offer an attractive alternate to warfarin for patients with nonvalvular atrial fibrillation. The lack of drug-to-drug interactions and need for coagulation monitoring parameters such as international normalized ratio and a short half life make therapeutic control and withdrawal in the event of bleeding easier. The side-effect profile, including elevation of liver enzymes, may temper the clinical utility of long-term use ximelagatran. PMID- 15821835 TI - Effect of flaxseed consumption on blood pressure, serum lipids, hemopoietic system and liver and kidney enzymes in healthy humans. AB - BACKGROUND: Effects of flaxseed on serum lipids have been studied in humans, but the results are variable. Flaxseed is ineffective in lowering blood pressure in rats. Its effect on blood pressure in humans is not known. It is also not known if long-term use of flaxseed in humans has deleterious effects on the hemopoietic system, serum glucose, and renal and kidney function. We investigated the effect of short-term use of flaxseed in humans on arterial pressure and serum lipids (triglycerides, total cholesterol; high-, low-, and very-low-density-lipoprotein cholesterol [HDL-C, LDL-C, VLDL-C], hemopoietic system (red blood cells, neutrophils, hemoglobin) and the various biochemical parameters, such as serum protein, albumin, total bilirubin, aspartate aminotransferase, alkaline phosphatase, creatinine, urea, related to hepatic and renal function, and serum glucose. METHODS: Fifteen healthy men, aged 22 to 47 years, consumed three muffins daily containing 32.7 g of total flaxseed for 4 weeks, in addition to their normal daily diet. Blood pressure and blood samples for various biochemical measurements were collected before and after 4 weeks of flaxseed diet. RESULTS: Blood pressures, heart rate, hemoglobin, and counts of red blood cells, white blood cells, and neutrophils remained unaltered after flaxseed diet. Serum total cholesterol, HDL-C, LDL-C, and VLDL-C remained unchanged, but serum triglycerides levels were elevated. Serum total bilirubin, aspartate aminotransferase, alkaline phosphatase, protein, albumin, glucose, and urea remained unaltered, but serum levels of creatinine decreased. CONCLUSION: These results suggest that 4 weeks use of flaxseed does not have deleterious effects on the hemopoietic system or renal and hepatic function and does not lower blood pressure and serum lipids. However, the level of serum triglyceride level was elevated. PMID- 15821836 TI - Long-term benefits and limitations of combined antianginal drug therapy in elderly patients with symptomatic chronic coronary artery disease. AB - BACKGROUND: Chronic angina is a common and disabling disorder in the elderly. Combined antianginal drug treatment represents the mainstay of therapy in this population. However, there is a paucity of data regarding the effect of this strategy on long-term outcome in the elderly. METHODS: To assess the long-term effect of combined antianginal drug therapy in elderly individuals, we performed a long-term follow-up analysis of all 148 patients of the Trial of Invasive versus Medical therapy in Elderly (TIME) patients with chronic symptomatic coronary-artery disease assigned to an optimized medical therapy strategy. Angina severity, measures of quality of life (QOL), and survival were assessed after a median of 3.7 (0.1-6.9) years. RESULTS: At baseline, patients were 79.8 +/- 3.5 years old with Canadian Cardiovascular Society (CCS) class angina 3.0 +/- 0.7 despite the use of 2.4 +/- 0.6 antianginal drugs. Although antianginal drugs were increased to 2.8 +/- 0.9 (P < .01), 63 (43%) patients needed revascularization for refractory symptoms during the first year of observation (REVASC). At baseline, REVASC patients had more frequently CCS class 4 angina (37% vs 20%, P < 0.05) but reported less prior heart failure (5% vs 20%, P < 0.01), fewer prior cerebral events (3% vs 13%, P < .05) and a lower rate of two or more comorbidities (10% vs 33%, P < .01) than patients on continued drug therapy (DRUG). At long-term follow-up, angina severity was still higher in DRUG compared to REVASC patients (CCS class, 1.8 +/- 1.6 vs 1.0 +/- 1.4, P < .05) despite more antianginal drugs (2.1 +/- 1.1 vs 1.5 +/- 1.0, P < .01), whereas measures of QOL had improved similarly in both groups. In addition, long-term mortality was significantly higher in DRUG than in REVASC patients (38% vs 13%, P < .01). CONCLUSION: Combined antianginal drug therapy successfully relieved symptoms in most elderly patients with chronic angina but failed to do so in 43%. Patients who needed revascularization for refractory symptoms reported less angina, despite lower drug use during long-term follow-up and had a better long-term survival. Thus, the widely used strategy to increase antianginal drug therapy in elderly patients instead of evaluating them for revascularization should be reconsidered. PMID- 15821837 TI - Trends in the care of patients with acute myocardial infarction at a university affiliated Veterans Affairs Medical Center. AB - BACKGROUND: Acute cardiac care of the veterans at Veterans Administration (VA) hospitals has been thought of as poor in quality. We examined the use of life saving, evidence-based medical therapy in patients admitted with acute myocardial infarction to the University of Arkansas for Medical Sciences-affiliated VA Medical Center in Little Rock and compared the use of this therapy with other hospitals in Arkansas and in the rest of the nation. METHODS: Use of life-saving medical therapy in 117 patients admitted with acute myocardial infarction from January 2002 to December 2002 was compared with the National Registry of Myocardial Infarction database for the identical period. RESULTS: Heparin/low molecular-weight heparin and glycoprotein IIb/IIIa inhibitors were used in 88% and 66% of patients, respectively. Aspirin, beta adrenergic-blocking agents, angiotensin-converting enzyme (ACE) inhibitors/angiotensin receptor blockers (ARBs), and 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) were used in 92%, 93%, 62%, and 79% of the patients, respectively. The use of these therapies was better than in similar patients in Arkansas (P < .001) and the United States as a whole (P < .01). Calcium-channel blockers were used in 16% of the patients. At a mean follow-up period of 1.5 years, use of beta blockers and aspirin had decreased, whereas the use of statins and ACE inhibitors/ARBs was unchanged. CONCLUSION: This study shows that patients with acute myocardial infarction admitted to this university-affiliated VA Medical Center receive evidence-based life-saving medical therapy more often than in the rest Arkansas or in the entire United States. More important, patients at this federal institution continue to receive life-saving medical therapy during follow-up. Better use of evidence-based therapy may be related to affiliation of this VA Medical Center with a teaching institution where board certified cardiologists are involved in short- and long-term care of these patients. PMID- 15821838 TI - Evaluation of methods for the measurement of low-density lipoprotein cholesterol. AB - A high concentration of low-density lipoprotein cholesterol (LDL-C) in plasma is one of the strongest risk factors for atherosclerotic cardiovascular disease and mortality. The most common approach to determining LDL-C in the clinical laboratory is the Friedewald calculation. There is an increased interest to improve the accuracy of LDL-C estimated by this equation. The expert panel convened by National Cholesterol Education Program has recommended the development of accurate direct methods to measure LDL-C. Several homogeneous and fully automated methods have been introduced in recent years that show improved precision and accuracy over earlier methods, especially the Friedewald calculation. Each of the atherogenic particles in plasma--very-low, intermediate , and low- density lipoprotein--as well as lipoprotein (a), contain one molecule of apolipoprotein B (apoB) and thus, plasma total concentration of apoB reflects the number of atherogenic particles. Several studies suggested that the measurement of apoB could improve the prediction of risk of coronary artery disease. Thus, in addition to the newly developed direct assays, alternative calculation procedures have been proposed that also take into consideration total serum apoB concentration for the estimation of LDL-C and the presence of small, dense LDL particles. The new generation of homogenous methods for the measurement of LDL-C and the use of serum apoB concentration for the estimation of LDL-C can contribute to the accurate LDL-C determination. PMID- 15821839 TI - Administration of vascular endothelial growth factor adjunctive to fetal cardiomyocyte transplantation and improvement of cardiac function in the rat model. AB - BACKGROUND: The functional impact of cellular transplantation and the potential role of the addition of angiogenic factors for survival of engrafts remain controversial. METHODS: Vascular endothelial growth factor (VEGF) (25 ng/mL) was added to cultured fetal cardiomyocytes labeled with bromodeoxyuridine (BrDU), which was injected into the border zones of myocardial infarction 4 weeks after coronary occlusion in rat hearts. Group 1 (n = 12) received cells plus VEGF protein (100 ng), group 2 (n = 12) received cells without VEGF, group 3 (n = 10) received VEGF without cells, and group 4 (n = 12) received pure culture medium. Cardiac function was then assessed by transthoracic two-dimensional echocardiography and Langendorff perfusion system. In situ hybridization for Y chromosomes of transplanted cells, detection of BrDU-labeled cells, and platelet/endothelial cell adhesion molecule-1 (PECAM-1) staining for endothelial cells was performed. RESULTS: Echocardiography revealed smaller end-diastolic left ventricular dimensions in transplanted hearts in group 1 (0.83 +/- 0.13 cm 4 weeks after coronary occlusion before transplantation and 0.69 +/- 0.14 cm 2 months after transplantation, P < .05) and in group 2 (0.88 +/- 0.09 cm after coronary occlusion before transplantation and 0.76 +/- 0.08 cm 2 months after transplant), and increases in fractional shortening (34.2% +/- 8.53% before transplant and 45.3% +/- 10.9% after [P < .05] in group 1; 26.9% +/- 6.02% before transplant and 37.15% +/-8.08% after [P < .005] in group 2), whereas groups 3 and 4 showed a decrease in fractional shortening. Transplanted hearts developed higher pressures at rest (group 1, 96.8 +/- 20.8 mm Hg; group 2, 98.6 +/- 21.9 mm Hg) compared with controls (group 4, 70.9 +/- 25 mm Hg) (P < .05) and during inotropic stimulation (group 1, 111 +/- 19.5 mm Hg and group 2, 113.3 +/- 32.6 vs group 4, 80.7 +/- 31.6 mm Hg, P < .05). Histologic analysis demonstrated the presence of transplanted cells in border zones of infarcted host myocardium with neovascularization in all transplanted hearts. CONCLUSION: Transplantation of fetal cardiomyocytes results in improvement of left ventricular function. The addition of VEGF does not contribute to further improvement of left ventricular function and angiogenesis in the present model. PMID- 15821840 TI - The influence of extracellular acidosis on the effect of IKr blockers. AB - BACKGROUND: Myocardial infarction causes the acidification of the cellular environment and the resultant acidosis maybe arrhythmogenic. The effect of acidosis on the action of antiarrhythmic drugs, an important issue in the antiarrhythmic drug therapy after myocardial infarction, remains to be studied. METHODS: To evaluate the effect of acidosis on rectifier potassium current (Ikr) blockers, the human ether-a-go-go-related gene (HERG), which encodes IKr, was expressed in Xenopus laevis oocytes. The two electrodes voltage clamp technique was used and the experiments were performed at room temperature. RESULTS: Quinidine (10 microM) inhibited HERG tail current by 37% +/- 5% at pH7.4. The block decreased to 5% +/- 2% with extracellular pH at 6.2. Dofetilide (0.3 microM) inhibited HERG tail current by 34% +/- 3% and 1% +/- 2% at extracellular pH 7.4 and 6.2, respectively. Azimilide (10 microM) inhibited HERG tail current by 59% +/- 3% and 17% +/- 3% at extracellular pH 7.4 and 6.2. There were significant differences in the HERG inhibition by quinidine, dofetilide, and azimilide between pH 7.4 and pH 6.2 (P < .01). The drug concentration blocking 50% of current (IC50) was 5.8 +/- 0.3 microM for azimilide, 9.9 +/- 1.0 microM for quinidine, and 0.5 +/- 0.02 microM for dofetilide at pH 7.4. When extracellular pH was decreased from 7.4 to 6.2, the IC50 increased to 95.5 +/- 11.3 microM for azimilide, 203.2 +/- 15.7 microM for quinidine, and 12.6 +/- 1.2 microM for dofetilide. Unlike quinidine, dofetilide, and azimilide, there was no significant difference in the percentage of current block by amiodarone between pH 6.2 and 7.4. For amiodarone, the IC50 was 38.3 +/- 8.5 microM at pH 7.4 and 27.3 +/- 1.6 microM at pH 6.2. CONCLUSION: Our data show that the Ikr blocking effect of azimilide, dofetilide, and quinidine was attenuated at acid pH, whereas this was not the case for amiodarone. These observations may explain the efficacy of amiodarone in reducing arrhythmic death in patients after a myocardial infarction compared with other IKr blockers. PMID- 15821841 TI - Successful treatment of hypotension associated with stunned myocardium with oral midodrine therapy. AB - Myocardial stunning, a reversible decrease in the contractile function of the myocardium after an ischemic insult, often leads to hypotension that requires therapy with intravenous inotropes. We used the oral agent midodrine to treat hypotension that complicated myocardial stunning after successful revascularization with percutaneous coronary intervention in the setting of myocardial infarction and ischemia. Oral midodrine may offer a useful substitute to intravenous inotropic therapy and can shorten the duration of intensive care unit and hospital stay in this setting. PMID- 15821842 TI - Army pediatricians in Iraq. PMID- 15821843 TI - Review of thrombophilic States. PMID- 15821844 TI - Sleep-related disorders in ADHD: a review. PMID- 15821845 TI - Physician knowledge and management of children exposed to domestic violence in Ohio: a comparison of pediatricians and family physicians. AB - Active members of the Ohio chapters of American Academy of Family Physicians (FP=1,498) and American Academy of Pediatrics (Ped=1,725) were surveyed about their knowledge and management regarding children exposed to domestic violence (DV). Characteristics of respondents were analyzed by use of Chi-square analysis. Logistic regression was performed to identify predictors of DV knowledge and management. The response rate was 33.3%. Family physicians were more likely to know their local DV agency and recognize the adult symptoms of DV, such as unexplained injury. Pediatricians were more likely to report the child who saw a fight between parents to child protective services. Continuing work to increase physicians' comfort and ability to assess for DV and manage exposed children is needed. PMID- 15821846 TI - Variation in establishing a diagnosis of obesity in children. AB - Consensus guidelines provide recommendations for the diagnosis and management of obesity. We conducted a medical record review of children initially diagnosed with obesity at a general pediatrics visit. The diagnosis was made most often at health maintenance visits (46%). Body mass index was documented in 5% of initial visits; 74% had documentation of obesity-related history; 64% had documentation of counseling. In multivariate analysis, male patients were more likely to have diet history documentation; female patients were more likely to have weight loss program referrals. Future research should assess pediatricians' perceptions about obesity to better understand clinical practice patterns. PMID- 15821847 TI - Clinical utility of the bispectral index score when compared to the University of Michigan Sedation Scale in assessing the depth of outpatient pediatric sedation. AB - This single blinded observational study compared the bispectral index (BIS) monitor with a validated pediatric sedation scale, the University of Michigan Sedation Scale (UMSS), to evaluate whether the BIS score can be used to judge the depth of sedation in pediatric outpatients. Thirty-eight children, with a mean age of 5.8 years, undergoing routine sedation for both noninvasive and gastrointestinal procedures, had simultaneous BIS and UMSS scores recorded. Sedation categories were defined as light, moderate, and deep for both UMSS and BIS. There was a moderate correlation between BIS and the UMSS, Spearman's r < 0.499. The correlation was poor for children receiving chloral hydrate, r < 0.213. The BIS score was not predictive of any specific UMSS score. The UMSS and BIS categories of sedation matched only 36% of the time. BIS underestimated the clinical level of sedation. PMID- 15821848 TI - Factitious disorder and malingering in adolescent girls: case series and literature review. AB - Six cases of factitious disease and malingering in pediatric patients referred to an infectious diseases practice in a tertiary care children's hospital are described, and implications for general clinical practice are reviewed. All patients were girls aged 9-15 years. Two patients were malingering with the secondary gain of avoiding attendance at school. The other 4 patients presented with factitious illness without clear link to secondary gain, but rather for a psychological purpose. Three of the subjects admitted to self-induced or feigned illness. The 2 patients diagnosed with malingering did very well with early parental support, psychotherapy, and attention paid to school difficulties. The outcomes of the others with underlying psychological conflicts were less resolved. Factitious disorders and malingering occur in the pediatric population. A high index of suspicion is needed for prompt diagnosis and care. PMID- 15821849 TI - Treatment of labial fusion with topical estrogen therapy. AB - Twenty girls with labial fusion that covered at least 50% of the vaginal opening were treated with a topical estrogen cream. The cream was precisely applied to the fused area twice a day until the adhesions were totally lysed. After separation of the adhesions, a petroleum ointment (Vaseline) was applied twice a day to the labia minora for at least 1 month. The introital area was carefully rinsed twice each day before application of either the estrogen cream or the petroleum ointment. The mean age of the studied population was 13.2 months (range, 2 to 38 months). The mean duration of estrogen treatment was 2.4 months (range, 1 to 3.5 months). All patients were successfully treated. Five patients developed vulval pigmentation, which lasted for a mean of 1.5 months. One patient developed breast enlargement, which lasted for 1 month. The mean duration of follow-up was 3.1 months, and there was no recurrence of labial fusion during the period of follow-up. We conclude that treatment of labial fusion with topical estrogen therapy is safe and effective. PMID- 15821850 TI - Presentation of pediatric celiac disease in the United States: prominent effect of breastfeeding. AB - Childhood celiac disease (CD) is considered rare in the United States. Consequently there are few data concerning its clinical presentation. A validated questionnaire was distributed to families of children with CD. One hundred forty one children with biopsy-proven CD were included in the study. We found significant differences in the clinical spectrum of children based on their infant feeding history. Exclusively breastfed children were significantly less likely to report failure to thrive (69% vs 88%, p<0.05) and short stature (37% vs 62%, p<0.05), and had a higher rate of "atypical'' symptoms (p<0.01). Breastfeeding alters the presentation and contributes to atypical presentations of CD and diagnostic delay. Pediatricians need to be aware of the diverse manifestations of celiac disease to reduce diagnostic delay. PMID- 15821851 TI - Subacute sclerosing panencephalitis in a patient with cerebral palsy. PMID- 15821852 TI - Pulmonary arteriovenous malformation: an unusual case of hypoxemia in an infant. PMID- 15821853 TI - Neonatal hyperparathyroidism due to maternal hypoparathyroidism and vitamin D deficiency: a cause of multiple bone fractures. PMID- 15821854 TI - A swollen neck. PMID- 15821856 TI - Respiratory distress. PMID- 15821857 TI - Monitoring dioxins in food and feedstuffs using accelerated solvent extraction with a novel integrated carbon fractionation cell in combination with a CAFLUX bioassay. AB - The concentrations of dioxins in fish oil and fish meal were determined with accelerated solvent extraction, using a novel integrated carbon fractionation extraction cell followed by a miniturized multilayer silica column and bioanalysis on a recently-developed chemically-activated fluorescent gene expression cell bioassay. The developed method allows for simultaneous gravimetric lipid weight determination, which was shown for both matrices under study (about 100% lipid recovery of each sample). Initial results practically meet the quality criteria on screening methods for control of dioxins in food and feedstuffs laid down in the EU Commission Directives 2002/69/EC (food) and 2002/70/EC (feed). This demonstrates that the developed method can be used as a screening tool for monitoring dioxins in food and feed after some additional improvements and testing on a greater number of matrices. PMID- 15821858 TI - Surface potential mapping of dispersed proteins. AB - We describe a method for detecting proteins after transfer to PVDF membranes, based on the surface potential attributed to each protein. Proteins separated by classical two-dimensional polyacrylamide gel electrophoresis could be detected by scanning the membrane surface with a vibrating capacitor (also called a Kelvin probe) on the basis of differences between their surface potential and that of the membrane. Coupled to colloidal gold staining, the technique enables detection of proteins previously undetectable by classical staining methods. Plotting variations of the surface potential in two dimensions visualizes proteins which migrate close together. Finally, we demonstrate that the Kelvin probe detects proteins over a concentration range from micro to sub-nanogram with increased sensitivity at lower concentrations, and unlike other methods, appears to be similar for all proteins tested so far. The method described is fast, reliable, and it can be automated for high throughput. PMID- 15821859 TI - Laudatio Norman M. Rich. PMID- 15821860 TI - The deep belly of the temporalis muscle: an anatomical, histological and MRI study. AB - In order to achieve a better functional and clinical knowledge of a masticatory muscle called the sphenomandibularis that is suspected to be responsible for headaches by compressing the maxillary nerve, bilateral dissections of the infratemporal fossa were performed on ten human cadavers and completed by histological and radiological studies of the same areas. Both macroscopic and microscopic observations obviously showed that the so-called sphenomandibularis muscle corresponds to the deep portion of the temporalis muscle, since there is no epimysial septum between these two structures, which previously have been described as being completely independent from each other. In spite of the close topographic relationship between the deep belly of the temporalis and the lateral pterygoid muscle, as well as their similar innervation pattern, the sphenomandibularis in fact has to be considered functionally as an original but non-isolated positional fascicle of the temporalis muscle itself. Our observations, correlated with MR images, suggest indeed that the deep belly of the temporalis muscle is of functional importance in the masticatory movements, but is not involved by its neurovascular vicinity in the genesis of specific headaches. Its surgical release, however, should be discussed in the case of a temporal myoplasty. PMID- 15821861 TI - Inguinal hernia repair: results using an open preperitoneal approach. AB - BACKGROUND: Laparoscopic surgical approaches to the repair of inguinal hernias have shown the advantages of placing mesh in the preperitoneal space. Despite those advantages, laparoscopic hernia repairs have been associated with increased cost, longer operating times, and advanced laparoscopic skills. An open preperitoneal approach has the benefit of mesh in the preperitoneal position without the disadvantages of a laparoscopic procedure. METHODS: We present our experience with the use of an open preperitoneal mesh repair (KugelMesh, Bard, Inc.). The study was conducted in a prospective fashion from January 1998 through October 2001. 1072 hernias were repaired in two community hospitals by three general surgeons. Patients with recurrent hernias were excluded if the initial repair was from a preperitoneal approach. Operative time, cost, post-operative pain, and complications were analyzed. RESULTS: Recurrences occurred in five patients (0.47%) during a mean follow-up time of 23 months (range: 2-47). The average operating time was 32.4 min (range: 16-62). Post-operative narcotic pain medication usage averaged 5.8 pills (range: 0-26) per repair. Average surgical charges were less for the open preperitoneal approach ($2253) than for laparoscopic repairs ($4826). CONCLUSIONS: The open preperitoneal hernia repair using the Kugel mesh offers many advantages. It is inexpensive, has a low recurrence rate, and allows the surgeon to cover all potential defects with one piece of mesh. Postoperative recovery is short and postoperative pain is minimal. PMID- 15821862 TI - Neurocysticercosis presenting as sudden death. AB - We report an acute case of a native woman from Peru suffering with cephalalgia which rapidly worsened and ended in her sudden and unexpected death. Brain CT scan showed hydrocephalus and intraventricular calcifications. Histopathological study revealed the presence of intraventricular taenia solium cysts which caused blockage of cerebrospinal fluid and secondary hydrocephalus. Due to the increasing travel movements of people neurocysticercosis must be considered as a cause of unexplained sudden death. PMID- 15821863 TI - Haemorrhagic complications and the incidence of asymptomatic bleeding associated with stereotactic brain biopsies. AB - OBJECT: Stereotactic brain biopsy is a routinely used technique for the diagnosis of brain lesions. Due to its minimally invasive nature, the potential risks associated with this procedure are sometimes underestimated. We have retrospectively analyzed the incidence of symptomatic and asymptomatic haemorrhagic complications associated with stereotactic biopsies. Various variables that may contribute to such complications have been retrospectively analyzed. METHODS: Medical and radiological records of 355 consecutive patients who underwent a diagnostic stereotactic brain biopsy were reviewed. The incidence of haemorrhage was derived from a routine post-operative CT scan done within 90 120 minutes of the biopsy. Demographic, radiographic, pathological, and clinical data were also extracted and evaluated for their possible association with haemorrhagic complications. RESULTS: Twenty-five patients (7%) experienced haemorrhagic complications associated with stereotactic biopsy, about half of whom (3.4%) were asymptomatic with no impact on the clinical course. Thirteen (3.6%) complications were symptomatic and two patients (0.6%) died. Lesions located in the brainstem were found to have a significantly higher rate of complications compared to other locations. No other variables, such as location, edema, number of biopsy specimens, or pre-existing neurological deficit showed a statistically significant impact on the incidence or severity of haemorrhage. Seven of the symptomatic complications occurred immediately post biopsy, but in six patients they developed within several hours and even days. The overall diagnostic yield of the biopsies was 93.8%, but was somewhat lower in patients experiencing a haemorrhagic complication. CONCLUSIONS: Stereotactic brain biopsy was associated with a low incidence of symptomatic haemorrhagic complications, morbidity and mortality, and a high diagnostic yield. About half of the haemorrhagic complications were asymptomatic. Lesions located in the brainstem had a higher rate of complications. No other clinical, radiographic, or pathological variables were found as predictors of increased risk for haemorrhage. PMID- 15821864 TI - DNA binding and pairing activity of OsDmc1, a recombinase from rice. AB - A cloned cDNA corresponding to OsDMC1 from rice anther tissue was expressed in Escherichia coli. The OsDmc1 protein was largely present in the inclusion bodies of the cell lysatE., which was solubilized by 8.0 M urea containing buffeR., purified to homogeneity by Ni-CAM agarose column chromatography, followed by renaturation to its native state through stepwise dialysis against reduced concentrations of urea. The purified protein cross-reacted with anti-yeast Dmc1 antibodies. The binding efficiency observed with circular single-stranded DNA (ssDNA) was similar to that with circular double-stranded DNA (dsDNA). The binding to either DNA showed no ATP dependencE., but required 5-10 mM Mg2+ in the presence of ATP. Even though the protein binding to dsDNA was as efficient as it was to ssDNA, the former induced no DNA dependent ATPasE., whereas the binding to ssDNA stimulated a significant level of DNA dependent ATPase activity. OsDmc1 ssDNA complex, with its ATPase proficiency, also mediated renaturation of homologous complementary strands as well as assimilation of single strands into homologous supercoiled duplexes leading to D-loop formation. The D-loop formation was lowered by excess of OsDmc1 protein. This D-loop formation activity was promoted by non-hydrolyzable ATP analog, AMP-PNP and was not observed in absence of ATP or presence of ADP/ATP-gamma-S. These properties reflected the classical hallmarks of a recombinase and represented the first biochemical characterization of a plant Dmc1 protein. PMID- 15821865 TI - cDNA cloning, functional expression and antifungal activities of a dimeric plant defensin SPE10 from Pachyrrhizus erosus seeds. AB - SPE10 is an antifungal protein isolated from the seeds of Pachyrrhizus erosus. cDNA encoding a 47 amino acid peptide was cloned by RT-PCR and the gene sequence proved SPE10 to be a new member of plant defensin family. The synthetic cDNA with codons preferred in yeast was cloned into the pPIC9 plasmid directly in-frame with the secretion signal alpha-mating factor, and highly expressed in methylotrophic Pichia pastoris. Activity assays showed the recombinant SPE10 inhibited specifically the growth of several pathogenic fungi as native SPE10. Circular dichroism and fluorescence spectroscopy analysis indicated that the native and recombinant protein should have same folding, though there are eight cystein residues in the sequence. Several evidence suggested SPE10 should be the first dimeric plant defensin reported so far. PMID- 15821866 TI - Transcriptional regulation by abscisic acid in barley (Hordeum vulgare L.) seeds involves autoregulation of the transcription factor HvABI5. AB - The barley bZIP transcription factor HvABI5 mediates abscisic acid (ABA) upregulated gene expression in barley (Hordeum vulgare L.) seeds. HvABI5 specifically recognizes cis-elements of the ABA response complexes present in the promoters of the ABA-induced genes HVA1 and HVA22. HvABI5 together with another transcription factor, HvVP1, are required for the transactivation of these promoters, and this transactivation process is insensitive to the negative regulator abi1-1. The expression of HvABI5 itself appeared to be induced by ABA and can be suppressed by abi1-1. Gain- and loss-of-function studies in barley aleurone cells show that HvABI5 expression is positively regulated by a feed forward circuit that involves HvABI5 itself and HvVP1. Mutation of the Ser residue in HvABI5, which has been shown to be phosphorylated in an ABA-dependent manner in the rice orthologue of HvABI5, reduces the transactivation activity of the factor by 50%. Although levels of HvABI5 and its transcript are enhanced by ABA treatment, the nuclear localization of HvABI5 is not affected by ABA. A model based on these observations is presented to explain the ABA upregulation of gene expression. PMID- 15821867 TI - The complex developmental expression of a novel stress-responsive barley Ltp gene is determined by a shortened promoter sequence. AB - The search for a cereal promoter capable of driving preferential transgene expression in the pericarp epidermis (epicarp) of developing barley (Hordeum vulgare L.) resulted in the cloning of a novel gene. This encoded a polypeptide of 124 amino acids showing 87 identity with WBP1A, a wheat lipid transfer protein (LTP), but much lower homology to other barley LTPs. In addition to the epicarp, this Ltp-like gene, Ltp6, is highly expressed in coleoptiles and embryos under normal growth conditions. Messenger RNA levels increased in seedling tissues during salt and cold treatments and under applied abscisic acid (ABA) and salicylic acid (SA). Taken together, Ltp6 tissue-specific and response patterns are distinct from other known barley Ltp genes. Inverse PCR was used to derive 2345 bp of upstream Ltp6 sequence. The level of transcription conferred by different promoter deletion constructs was assessed by quantitative real time RT PCR using gfp as a reporter in transient expression assays. All constructs containing at least 192 bp of upstream sequence and the 5'UTR conferred tissue specific expression and retained most of the promoter strength. Deletion of 64 bp (-192/-128) from this upstream sequence reduced expression levels by 80. Moreover, a minimal 247 bp Ltp6 promoter continuously drove gfp expression during spike development, from early ovary differentiation through its final expression in the epicarp and during embryogenesis and germination in transgenic barley, reproducing the expression pattern of the native gene. The potential use of this promoter sequence for targeting transgene-mediated disease resistance in barley and wheat is discussed. PMID- 15821868 TI - Expression of glutathione-S-transferase and its role in plant growth and development in vivo and shoot morphogenesis in vitro. AB - The enzymes glutathione-S-transferases (GSTs, E.C.2.5.1.18) have been associated with detoxification of xenobiotics, limiting oxidative damage and other stress responses in plants. In this study, we report the isolation of a mustard gene, BjGSTF2, homologous to the phi class GSTs and changes in plant growth in vivo and shoot regeneration in vitro were related to GST expression. GST transcripts accumulated differentially in mustard organs, where transcript was most abundant in root. Tissues incubated at high temperature or in the presence of exogenous H2O2, HgCl2, 1-aminocyclopropane-1-carboxylate, salicylic acid and paraquat upregulated GST expression, whereas spermidine was inhibitory. To investigate the in vivo function of GST, transgenic Arabidopsis thalianaplants expressing sense (GST-S6), antisense (GST-A4) and double-stranded BjGSTF2 (GST-DS1) RNAs were generated. GST-S6 was shown to flower two days earlier and was relatively more tolerant to HgCl2 and paraquat, whereas GST-DS1 with least stress tolerance flowered one week later compared to WT and GST-A4. In shoot regeneration response, tissues originated from GST-S6 were highly regenerative, whereas no shoot regeneration was observed in GST-DS1 tissues after 30 days of culture. Results of this study provide the evidence showing that GST plays a role in plant growth and development in vivo and shoot regeneration in vitro. PMID- 15821869 TI - Two WD-repeat genes from cotton are functional homologues of the Arabidopsis thaliana TRANSPARENT TESTA GLABRA1 (TTG1) gene. AB - Cotton fibres are single, highly elongated cells derived from the outer epidermis of ovules, and are developmentally similar to the trichomes of Arabidopsis thaliana. To identify genes involved in the molecular control of cotton fibre initiation, we isolated four putative homologues of the Arabidopsis trichome associated gene TRANSPARENT TESTA GLABRA1 (TTG1). All four WD-repeat genes are derived from the ancestral D diploid genome of tetraploid cotton and are expressed in many tissues throughout the plant, including ovules and growing fibres. Two of the cotton genes were able to restore trichome formation in ttg1 mutant Arabidopsis plants. Both these genes also complemented the anthocyanin defect in a white-flowered Matthiola incana ttg1 mutant. These results demonstrate parallels in differentiation between trichomes in cotton and Arabidopsis, and indicate that these cotton genes may be functional homologues of AtTTG1. PMID- 15821870 TI - Transcriptional activation of tobacco E2F is repressed by co-transfection with the retinoblastoma-related protein: cyclin D expression overcomes this repressor activity. AB - Evidence is emerging that the E2F family of transcription factors plays an important role in the regulation of gene expression at the G1/S transition in plants. Here, we show that in the tobacco proliferating cell nuclear antigen (PCNA), whose transcript is specifically expressed at G1/S phase, the two E2F binding sites are synergistically responsible for transcriptional activation at G1/S phase in synchronized tobacco BY-2 cells transformed with promoter constructs fused to a reporter gene. In addition, we have isolated the tobacco DP cDNA (NtDP) and showed that significant activation of the reporter gene was observed in transient expression assays by concomitantly transfecting with plasmids expressing NtE2F and NtDP. This transcriptional activation was repressed by co-transfection with a plasmid expressing NtRBR1; in vitro pull-down assays also revealed that NtRBR1 binds directly to NtE2F, thereby potentially blocking the transcriptional activation of NtE2F. Importantly, this repressor activity was cancelled when NtRBR1 was further co-transfected with a plasmid expressing cyclin D but not with cyclin A or cyclin B. These results are discussed with respect to the repression activity of NtRBR1 on the NtE2F/NtDP complex. PMID- 15821871 TI - Recombinant human acid beta-glucosidase stored in tobacco seed is stable, active and taken up by human fibroblasts. AB - Gaucher disease, the most common genetic lysosomal disorder, is caused by the lack of functional acid beta-glucosidase (GCase) and is currently treated at a very high cost by enzyme replacement therapy. In an attempt to provide a safe and cost-effective production system, human placental GCase was produced and purified from transgenic tobacco seeds. Plant-derived recombinant GCase was found to be enzymatically active, uptaken by human fibroblasts and free of immunogenic xylose and fucose residues. This report demonstrates the potential of plant bioreactors in the large-scale production of injectable proteins required for lifelong therapy. PMID- 15821872 TI - A novel class of Helitron-related transposable elements in maize contain portions of multiple pseudogenes. AB - We recently described a maize mutant caused by an insertion of a Helitron type transposable element (Lal, S.K., Giroux, M.J., Brendel, V., Vallejos, E. and Hannah, L.C., 2003, Plant Cell, 15: 381-391). Here we describe another Helitron insertion in the barren stalk1 gene of maize. The termini of a 6525 bp insertion in the proximal promoter region of the mutant reference allele of maize barren stalk1 gene (ba1-ref) shares striking similarity to the Helitron insertion we reported in the Shrunken-2 gene. This insertion is embedded with pseudogenes that differ from the pseudogenes discovered in the mutant Shrunken-2 insertion. Using the common terminal ends of the mutant insertions as a query, we discovered other Helitron insertions in maize BAC clones. Based on the comparison of the insertion site and PCR amplified genomic sequences, these elements inserted between AT dinucleotides. These putative non-autonomous Helitron insertions completely lacked sequences similar to RPA (replication protein A) and DNA Helicases reported in other species. A blastn analysis indicated that both the 5' and 3' termini of Helitrons are repeated in the maize genome. These data provide strong evidence that Helitron type transposable elements are active and may have played an essential role in the evolution and expansion of the maize genome. PMID- 15821873 TI - Unique and overlapping expression patterns of Arabidopsis CYP85 genes involved in brassinosteroid C-6 oxidation. AB - Brassinosteroids (BRs) are steroid hormones that are essential for plant growth and development. To gain insight into potential sites of BR synthesis, we studied promoter activities of the two Arabidopsis BR C-6 oxidase genes (CYP85A1 and CYP85A2) in transgenic plants carrying promoter fusions with the GUS, GFP or LUC reporter genes. BR-dependent feedback regulation of the GUS reporter constructs indicated that their expression corresponded to those of the native genes. Both the CYP85A1 and CYP85A2 promoters showed maximum activity during the first week following germination, particularly in the vascular tissues. Compared to CYP85A2, CYP85A1 expression was weaker and confined to the early stages of seedling development. Stronger CYP85A2 promoter activity was evident in both juvenile and adult plants. Comparison of the 5'-UTR and TATA box sequences of CYP85A1 and CYP85A2 revealed high homology, indicating a relatively recent gene duplication. We also found that transgenic Arabidopsis plants harbouring the tomato DWARF promoter-GUS fusion had similarities in the expression pattern to the Arabidopsis genes suggesting common transcriptional regulation of CYP85 genes in the two species. PMID- 15821874 TI - Duplication of the S-locus F-box gene is associated with breakdown of pollen function in an S-haplotype identified in a natural population of self incompatible Petunia axillaris. AB - We previously identified both self-incompatible and self-compatible plants in a natural population of self-incompatible Petunia axillaris subsp. axillaris, and found that all the self-compatible plants studied carried either SC1- or SC2 haplotype. Genetic crosses showed that SC2 was identical to S17 identified from another natural population of P. axillaris, except that its pollen function was defective, and that the pollen-part mutation in SC2 was tightly linked to the S locus. Recent identification of the S-locus F-box gene (SLF) as the gene that controls pollen specificity in S-RNase-based self-incompatibility has prompted us to examine the molecular basis of this pollen-part mutation. We cloned and sequenced the S17-allele of SLF of P. axillaris, named PaSLF17, and found that SC2SC2 plants contained extra restriction fragments that hybridized to PaSLF17 in addition to all of those observed in S17S17 plants. Moreover, these additional fragments co-segregated with SC2. We used the SC2-specific restriction fragments as templates to clone an allele of PaSLF by PCR. To determine the identity of this allele, named PaSLFx, primers based on its sequence were used to amplify PaSLF alleles from genomic DNA of 40 S-homozygotes of P. axillaris, S1S1 through S40S40. Sequence comparison revealed that PaSLFx was completely identical with PaSLF19 obtained from S19S19. We conclude that the S-locus of SC2 contained both S17-allele and the duplicated S19-allele of PaSLF. SC2 is the first naturally occurring pollen-part mutation of a solanaceous species that was shown to be associated with duplication of the pollen S. This finding lends support to the proposal, based on studies of irradiation-generated pollen-part mutants of solanaceous species, that duplication, but not deletion, of the pollen S, causes breakdown of pollen function. PMID- 15821875 TI - Differential combinatorial interactions of cis-acting elements recognized by R2R3 MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes. AB - Chalcone synthase (CHS), chalcone flavanone isomerase (CFI), flavanone 3 hydroxylase (F3H) and flavonol synthase (FLS) catalyze successive steps in the biosynthetic pathway leading to the production of flavonols. We show that in Arabidopsis thaliana all four corresponding genes are coordinately expressed in response to light, and are spatially coexpressed in siliques, flowers and leaves. Light regulatory units (LRUs) sufficient for light responsiveness were identified in all four promoters. Each unit consists of two necessary elements, namely a MYB recognition element (MRE) and an ACGT-containing element (ACE). C1 and Sn, a R2R3 MYB and a BHLH factor, respectively, known to control tissue specific anthocyanin biosynthesis in Z. mays, were together able to activate the AtCHS promoter. This activation of the CHS promoter required an intact MRE and a newly identified sequence designated R response element (RREAtCHS) containing the BHLH factor consensus binding site CANNTG. The RRE was dispensable for light responsiveness, and the ACE was not necessary for activation by C1/Sn. These data suggest that a BHLH and a R2R3-MYB factor cooperate in directing tissue-specific production of flavonoids, while an ACE-binding factor, potentially a BZIP, and a R2R3-MYB factor work together in conferring light responsiveness. PMID- 15821876 TI - Specific interactions between Dicer-like proteins and HYL1/DRB-family dsRNA binding proteins in Arabidopsis thaliana. AB - Proteins that specifically bind double-stranded RNA (dsRNA) are involved in the regulation of cellular signaling events and gene expression, and are characterized by a conserved dsRNA-binding motif (dsRBM). Here we report the biochemical properties of nine such gene products, each containing one or two dsRBMs: four Arabidopsis Dicer-like proteins (DCL1-4), Arabidopsis HYL1 and four of its homologs (DRB2, DRB4, DRB5 and OsDRB1). DCL1, DCL3, HYL1 and the four HYL1 homologs exhibit significant dsRNA-binding activity, indicating that these proteins are involved in RNA metabolism. The dsRBMs from dsRBM-containing proteins (dsRBPs) also function as a protein-protein interaction domain and homo- and heterodimerization are essential for biological functioning of these proteins. We show that DRB4 interacts specifically with DCL4, and HYL1 most strongly interacts with DCL1. These results indicate that each HYL1/DRB family protein interacts with one specific partner among the four Dicer-like proteins. Localization studies using GFP fusion proteins demonstrate that DCL1, DCL4, HYL1 and DRB4 localize in the nucleus, while DRB2 is present in the cytoplasm. Subcellular localizations of HYL1, DRB4, DCL1 and DCL4 further strengthen the notion that HYL1 and DCL1, and DRB4 and DCL4, exist as complexes. The presented data suggest that each member of the HYL1/DRB protein family may individually modulate Dicer function through heterodimerization with a Dicer-like protein in vivo. PMID- 15821877 TI - Multiple insect resistance in transgenic tomato plants over-expressing two families of plant proteinase inhibitors. AB - Protease inhibitors have been proposed as potential defense molecules for increased insect resistance in crop plants. Compensatory over-production of insensitive proteases in the insect, however, has limited suitability of these proteins in plant protection, with very high levels of inhibitor required for increased plant resistance. In this study we have examined whether combined used of two inhibitors is effective to prevent this compensatory response. We show that leaf-specific over-expression of the potato PI-II and carboxypeptidase inhibitors (PCI) results in increased resistance to Heliothis obsoleta and Liriomyza trifolii larvae in homozygote tomato lines expressing high levels (>1% the total soluble proteins) of the transgenes. Leaf damage in hemizygous lines for these transformants was, however, more severe than in the controls, thus evidencing a compensation response of the larvae to the lower PI concentrations in these plants. Development of comparable adaptive responses in both insects suggests that insect adaptation does not entail specific recognition of the transgene, but rather represents a general adaptive mechanism triggered in response to the nutritional stress imposed by sub-lethal concentrations of the inhibitors. Combined expression of defense genes with different mechanisms of action rather than combinations of inhibitors may then offer a better strategy in pest management as it should be more effective in overcoming this general adaptive response in the insect. PMID- 15821878 TI - Large-scale statistical analysis of secondary xylem ESTs in pine. AB - A computational analysis of pine transcripts was conducted to contribute to the functional annotation of conifer sequences. A statistical analysis of expressed sequential tags(ESTs) belonging the 7732 contigs in the TIGR Pinus Gene Index (PGI1.0) identified 260 differentially represented gene sequences across six cDNA libraries from loblolly pine secondary xylem. Cluster analysis of this subset of contigs resulted in five groups representing genes preferentially represented in one of the xylem samples (compression wood, plannings, root xylem, latewood) and one group containing mostly genes simultaneously present in compression and side wood libraries. To complement the sequence annotation, 27 cDNA clones representing selected transcripts were completely sequenced. Several genes were identified that could represent putative markers for xylem from different organs, at different stages of development. Several sequences encoding regulatory proteins were over-represented in root xylem as opposed to the other xylem samples. Some of them belonged to known families of plant transcription factors, but two genes were previously uncharacterized in plants. One transcript was homologous to the gene encoding the Smad4 interacting factor, a key co-activator in TGFbeta (transforming growth factor) signalling in animals. Thus, the digital analysis of pine ESTs highlighted a putative gene function of potentially broad interest but that has yet to be investigated in plants. More generally, this study showed that the application of numerical approaches to EST databases should be helpful in establishing priorities among genes to consider for targeted functional studies. Thus, we illustrated the potential of extracting information from conifer sequences already accessible through well-structured public databases. PMID- 15821879 TI - A novel glucosyltransferase involved in steroid saponin biosynthesis in Solanum aculeatissimum. AB - Steroidal saponins are widely distributed in many plant species. Their diverse structures have resulted in a wide range of applications, including drug and medicine production. It has been suggested that the nature of the non-saccharide and oligosaccharide portions of the saponin molecule both contribute to the properties of individual saponins. Despite numerous studies on the occurrence, chemical structure, and varying pharmaceutical activities of steroidal saponins, their biosynthesis pathway is poorly understood. Glycosylation is thought to be the final step in steroidal saponin biosynthesis and it is thought to be involved in regulating the biological activities of saponins. Isolation of the glycosyltransferases that catalyze the transfer of sugar molecules to steroidal compounds will help to clarify the mechanisms that produce diverse saponins and control their activities in plants. In this study, we obtained three cDNAs encoding putative glycosyltransferases from Solanum aculeatissimum. One of the three, SaGT4A showed UDP-glucosyltransferase activity. This is the first cloned glucosyltransferase involved in steroidal saponin biosynthesis. SaGT4A catalyzes the 3-O-glucosylation of steroidal sapogenins, such as diosgenin, nuatigenin, and tigogenin. This enzyme also glucosylates steroidal alkaloids, such as solanidine, solasodine, and tomatidine. Gene expression analysis revealed that the accumulation of SaGT4A transcripts showed a unique response to wounding stress indicating the involvement of SaGT4A in plant defense system. PMID- 15821880 TI - The mitochondrial branched-chain aminotransferase (AtBCAT-1) is capable to initiate degradation of leucine, isoleucine and valine in almost all tissues in Arabidopsis thaliana. AB - Plants are capable to de novo synthesize the essential amino acids leucine, isoleucine and valine. Studies in recent years, however, also revealed that plants have the potential to degrade leucine or may be all of the branched-chain amino acids. One of the enzymes participating in both biosynthesis and degradation is the branched-chain aminotransferase, which is in Arabidopsis thaliana encoded by a small gene family with six transcribed members. We have now studied the steady state mRNA levels by quantitative RT-PCR and promoter activities of these genes with promoter::glucuronidase reporter gene constructs in transgenic plants. The gene encoding the mitochondrial isoenzyme (Atbcat-1) is expressed in all tissues with predominant transcription in seedlings and leaves. Surprisingly the plastid located proteins (AtBCAT-2, -3 and -5) are expressed at rather low levels with only Atbcat-3 transcribed in all tissues. The most likely cytoplasmic-located AtBCAT-4 and AtBCAT-6 are mainly expressed in tissues associated with transport function and in meristematic tissues, respectively. A detailed characterization of the enzyme activity and substrate specificity of the mitochondrial AtBCAT-1 enzyme revealed the potential of this enzyme to initiate degradation of all branched-chain amino acids. In addition alpha-aminobutyrate and alpha-ketobutyrate as well as methionine and alpha-ketomethylthiobutyrate are identified as substrates. This suggests that AtBCAT-1 and potentially other members of this protein family may influence methionine levels and may play an important role in the metabolism of the nonprotein amino acid alpha aminobutyrate. The consequences of these substrate specificities for bioplastic production and methionine homeostasis are discussed. PMID- 15821882 TI - Stable expression of a defense-related gene in wheat epidermis under transcriptional control of a novel promoter confers pathogen resistance. AB - Tissue-specific or regulated expression of transgenes is desirable in order to prevent pleiotropic side effects of putatively harmful transgene products as well as loss of energy resources due to unnecessary accumulation of transgene products. Epidermis-specific expression would be useful for many defense-related genes directed against attack by fungal pathogens that enter the plant body by direct penetration through the epidermis. In an approach to enhance resistance of wheat to the powdery mildew fungus Blumeria graminis f.sp. tritici, a novel epidermis-specific promoter was developed and used for expression of two defense related genes. A 2.3 kb fragment of the wheat GstA1 promoter in combination with an intron-containing part of the wheat WIR1a gene was found to drive strong and constitutive transient expression in wheat epidermis. This promoter-intron combination was used for overexpression of oxalate oxidase 9f-2.8 and TaPERO peroxidase, two defense-related wheat genes expressed in inner leaf tissues. Expression studies of several transgenic lines by in situ oxalate-oxidase staining, RNA and protein blot analyses, as well as real-time PCR, demonstrated strong and constitutive transgene expression in the shoot epidermis. Transient as well as stable over-expression of the TaPERO peroxidase gene in wheat epidermis under the control of the GstA1i promoter resulted in enhanced resistance against Blumeria graminis f.sp. tritici, whereas oxalate-oxidase overexpression had no effect in either system. The data suggest that the wheat GstA1 promoter in combination with the WIR1a intron is useful for transgenic approaches to fungal disease resistance in cereals. PMID- 15821883 TI - Characterisation and expression of the pathway from UDP-glucose to UDP-xylose in differentiating tobacco tissue. AB - The pathway from UDP-glucose to UDP-xylose has been characterised in differentiating tobacco tissue. A xylogenic suspension cell culture of tobacco has been used as a source for the purification of the enzymes responsible for the oxidation of UDP-glucose to UDP-glucuronic acid and its subsequent decarboxylation to UDP-xylose. Protein purification and transcriptional studies show that two possible candidates can contribute to the first reaction. Most of the enzyme activity in the cultured cells could be accounted for by a protein with an Mr of 43 kDa which had dual specificity for UDP-glucose and ethanol. The cognate cDNA, with similarity to alcohol dehydrogenases (NtADH2) was expressed in E. coli to confirm the dual specificity. A second UDP-glucose dehydrogenase, corresponding to the monospecific form, ubiquitous amongst plants and animals, could not be purified from the tobacco cell cultures. However, two cDNAs were cloned with high similarity to the family of UDP-glucose dehydrogenases. Transcripts of both types of dehydrogenase showed highest expression in tissues undergoing secondary wall synthesis. The UDP-glucuronate decarboxylase was purified as polypeptides of Mr 87 and 40 kDa. Peptide fingerprinting of the latter polypeptide identified it as a form of UDP-glucuronate decarboxylase and functionality was established by expressing the cognate cDNA in E. coli. Expression of 40 kDa polypeptide and its corresponding mRNA was also found to be highest in tissues associated with secondary wall formation. PMID- 15821884 TI - Biochemical and molecular characterization of photosystem I deficiency in the NCS6 mitochondrial mutant of maize. AB - Interorganellar signaling interactions are poorly understood. The maize non chromosomal stripe (NCS) mutants provide models to study the requirement of mitochondrial function for chloroplast biogenesis and photosynthesis. Previous work suggested that the NCS6 mitochondrial mutation, a cytochrome oxidase subunit 2 (cox2) deletion, is associated with a malfunction of Photosystem I (PSI) in defective chloroplasts of mutant leaf sectors (Gu et al., 1993). We have now quantified the reductions of photosynthetic rates and PSI activity in the NCS6 defective stripes. Major reductions of the plastid-coded PsaC and nucleus-coded PsaD and PsaE PSI subunits and of their corresponding mRNAs are seen in mutant sectors; however, although the psaA/B mRNA is greatly reduced, levels of PsaA and PsaB (the core proteins of PSI) are only slightly decreased. Levels of the PsaL subunit and its mRNA appear to be unchanged. Tested subunits of other thylakoid membrane complexes--PSII, Cyt b6/f, and ATP synthase, have minor (or no) reductions within mutant sectors. The results suggest that specific signaling pathways sense the dysfunction of the mitochondrial electron transport chain and respond to down-regulate particular PSI mRNAs, leading to decreased PSI accumulation in the chloroplast. The reductions of both nucleus and plastid encoded components indicate that complex interorganellar signaling pathways may be involved. PMID- 15821881 TI - Comparative analysis of early embryonic sunflower cDNA libraries. AB - To gain information concerning cell functions and activities during sunflower embryogenesis, an expressed sequence tag (EST) approach was used to analyse gene expression in the early stages of sunflower embryos development. Confocal microscopy observations of whole-mounted embryos allowed us to identify precisely the major steps of the zygotic embryonic development. A time-course analysis was then employed to collect the embryonic material. Three cDNA libraries were constructed from microdissected embryos, and three other cDNA libraries were created using a classical day after pollination schedule. A total of 7106 ESTs were produced and assembled. The total number of putative different genes represents about 43.1 (3064 tentative contigs and singlets) of the analysed sequences. The unigenes that showed similarity to proteins with known or predicted functions (50.3) were classified into 15 different functional categories. The functional profiles were found to be quite similar for all studied embryo stages but statistical analysis revealed that successive and coordinate sets of genes are expressed at each embryonic stage. The analysis allowed us to identify abundant and differentially expressed genes at the early stages of embryos development as well as some putatively interesting genes, showing strong similarities with genes playing key roles in plant and animal embryogenesis. The data presented in this study not only provide a first global overview of the genes expression profile during sunflower embryogenesis but also represent an original and valuable tool for developmental genomics studies on exalbuminous dicots. PMID- 15821885 TI - Recent advances in dendritic cell biology. AB - Dendritic cells are professional antigen presenting cells that are central to the induction and regulation of immunity. This review discusses recent advances in the understanding of dendritic cell biology. PMID- 15821886 TI - Nuclear-targeting autoantibodies induced nuclear PARP cleavage accompanied by more pronounced decrease of peripheral white blood cells than Ro/SSA and La/SSB antigen-targeting autoantibodies. AB - Autoantibody production and leukocytopenia may be linked in patients with lupus erythematosus (LE). Unclear is the ability of different autoantibody species to induce apoptosis and cell loss. Laboratory routine analyses (white blood cell counts, autoantibody detection), and flow cytometry (annexin V, CD3, CD4, CD8) have been performed in 126 consecutive LE-patients. Nuclei of PBMC were investigated flow cytometrically for the presence of the 85 kDa poly-(ADP-ribose) polymerase (PARP) fragment. Peripheral total white blood cells (WBC), lymphocytes, T-cells, CD3+ CD4+, and CD3+ CD8+ cells were significantly decreased in patients with LE (P from 1.2 x 10(-14) to P < .0008). In the presence of either antinuclear (P from 1.2 x 10(-14) to P < .0008) or anti-dsDNA antibodies (P from 2.9 x 10(-12) to P < .007) were significantly diminished. Differences in cell numbers in LE patients with versus without anti-Ro/SSA were less pronounced: significant differences could be only obtained in lymphocytes and T-cells (P < .02). Anti-La/SSB antibodies were accompanied by significant increased leukocytes (P < .02). PARP cleavage (85 kDa) in nuclei was preferentially observed in cases with nuclear targeting autoantibodies. These results indicate that nuclear targeting autoantibodies are associated to lower peripheral blood cells counts than Ro/SSA, and La/SSB cytoplasmic targeting autoantibodies. This provides an explanation for the pathogenesis of cytopenias associated with SLE. PMID- 15821887 TI - Lower CD4+ T lymphocyte nadirs may indicate limited immune reconstitution in HIV 1 infected individuals on potent antiretroviral therapy: analysis of immunophenotypic marker results of AACTG 5067. AB - BACKGROUND: Although initiation of potent antiretroviral therapy (ART) has significantly improved immune perturbations in individuals with AIDS, it is unclear which factors are most important in determining the degree of immune reconstitution. METHODS: Whole blood was analyzed at baseline and week 12 in six groups of subjects (n = 81): those with acute or following immune reconstitution after Pneumocystis jirovecii (previously known as Pneumocystis carinii) pneumonia (PcP) (two groups) and cytomegalovirus (CMV) retinitis (two groups), HIV infection without AIDS (one group), and healthy volunteers (one group). Absolute CD4+ and CD8+ T lymphocytes, naive (CD45RA+) and memory (CD45RO+) CD4+ T lymphocytes and percentages of activated CD8+ T lymphocytes (CD8+/CD38+/HLA-DR), and CD28 expression on CD4+ and CD8+ T lymphocytes were enumerated. RESULTS: The reconstituted CMV group, which had a history of a lower CD4+ T lymphocyte nadir compared to the reconstituted PcP group (15 cells/mm(3) versus 48 cells/mm(3); p = .013), had significantly lower absolute CD4+, CD8+ and naive CD4+ T lymphocytes and a trend toward lower memory CD4+ T lymphocytes compared to the reconstituted PcP group. Moreover, no difference was noted between the reconstituted groups in the proportion of subjects with undetected HIV-1 RNA. The reconstituted subjects had significantly lower absolute CD4+, memory CD4+ and naive CD4+ T lymphocytes than the HIV-positive controls and a significantly higher percentage of activated CD8+ T lymphocytes with a lower percentage of CD8+CD28 expression than the HIV negative controls. CONCLUSION: The association of CD4+ T lymphocyte nadir with the extent of immune reconstitution in HIV-infected individuals suggests that HIV 1 may cause irreparable immune system damage despite potent ART. PMID- 15821888 TI - Flow cytometry evaluation of the T-cell receptor Vbeta repertoire among HIV-1 infected individuals before and after antiretroviral therapy. AB - HIV-1 infection leads to serious impairment of the immune system and perturbations in the T cell receptor Vbeta repertoire are also described. Immune reconstitution can be potentially achieved in response to HAART. In the present study 10 patients were investigated for the Vbeta pattern expression before and after six months of HAART. TCR were analyzed for T CD4+ and CD8+ subsets, separately, by flow cytometry, using a monoclonal antibody set of 24 different Vbeta chains. Compared to eight Brazilian healthy controls, no differences in Vbeta pattern of expression was observed for patients before or on antiretroviral therapy. Some chains such as Vbeta 3, 14, 16, 20 and 21.3 were over utilized by both T subsets, independently of HIV infection and/or antiretroviral treatment, differing from the ones described for individuals of other nationalities. However, when each patient was taken individually, particular alterations were detected for the Vbeta gene usage, compared to controls, for all individuals. After treatment, significant Vbeta usage changes were observed for seven patients. One or more chains on both T subsets were engaged in this process, defining a preferential oligoclonal profile for TCR repertoire distribution, after HAART. Although no pattern of specific Vbeta changes was detected in the circulating T cells, we cannot exclude that differential immune responses to HIV or other important antigens are being focused by these cells. PMID- 15821889 TI - Determinants of CD4 counts among HIV-negative Ethiopians: role of body mass index, gender, cigarette smoking, khat (Catha Edulis) chewing, and possibly altitude? AB - To study the determinants of CD4% and CD4 counts among HIV-negative Ethiopians, and to identify factors susceptible to explain the low CD4 counts observed among Ethiopian subjects. Cohort studies among factory workers in Akaki and Wonji, Ethiopia. Clinical and laboratory examinations, including determination of HIV serological status and T-cell subsets, were performed during follow-up visits every six months. In addition, micronutrients (retinol, carotenoids, tocopherol, transferrin receptor, and selenium) plasma concentrations were determined in a subset of 38 HIV-positive and 121 HIV-negative participants. HIV-negative participants with at least one CD4 count measurement were 157 females in Akaki, 203 males in Akaki, and 712 males in Wonji. CD4 counts were independently and positively associated with body mass index (through an increase in lymphocyte counts), female gender (through an increase in CD4%), cigarette smoking (through an increase in CD4%), khat chewing (through an increase in both lymphocyte counts and CD4%), and Akaki study site (through a large increase in lymphocyte counts compensating a decrease in CD4%). Intestinal parasitic infections were not associated with CD4% or CD4 counts. Retinol, carotenoids, and alpha-tocopherol plasma concentrations decreased with HIV infection and advancing immunosuppression, but were not associated with CD4 counts among HIV-negative subjects. Low body mass index among Ethiopians may have contributed to their overall low CD4 counts. Other factors remain to be elucidated. PMID- 15821890 TI - Reduced type 1 and type 2 cytokines in antiviral memory T helper function among women coinfected with HIV and HCV. AB - Bias in cytokine responses has been proposed as a contributing mechanism to pathogenesis in persistent HIV or hepatitis C virus (HCV) infections. We investigated whether coinfection with HCV modifies the profile of antigen specific cytokine secretion in women persistently infected with HIV compared to women with single HIV or HCV infection. The T helper response to HIV, HCV and cytomegalovirus (CMV) as a positive viral control was dominated by type 1 cytokines (interleukin- [IL] 2, interferon- [IFN] gamma and tumor necrosis factor [TNF] alpha), with IFN-gamma as the most abundantly secreted. IL-4, IL-5 and IL 10 were low in healthy controls and patients. Robust CMV-specific responses contrasted with curtailed HCV-specific responses in HCV-infected women. The overall anti-viral profile was dominated by Th1 cytokines even in coinfected women but both type 1 and type 2 responses were reduced in HIV-infected women and more extensively in women with HCV/HIV coinfection. PMID- 15821891 TI - Immunogenic properties of a chimeric plant virus expressing a hepatitis C virus (HCV)-derived epitope: new prospects for an HCV vaccine. AB - A vaccine against Hepatitis C virus (HCV) is urgently needed due to the unsatisfactory clinical response to current therapies. We evaluated the immunological properties of a chimeric Cucumber mosaic virus (CMV), a plant virus engineered to express on its surface a synthetic peptide derived from many HVR1 sequences of the HCV envelope protein E2 (R9 mimotope). Evidence was obtained that the chimeric R9-CMV elicits a specific humoral response in rabbits. Furthermore, in patients with chronic HCV infection, purified preparations of R9 CMV down-modulated the lymphocyte surface density of CD3 and CD8, and induced a significant release of interferon (IFN)-gamma, interleukin (IL)-12 p70 and IL-15 by lymphomonocyte cultures. Finally, an R9 mimotope-specific CD8 T-cell response, as assessed by intracellular IFN-gamma production, was achieved in the majority of the patients studied. Our results open up new prospects for the development of effective vaccines against HCV infection. Moreover, the wide edible host range of CMV makes the production of an edible vaccine conceivable. PMID- 15821892 TI - Bystander effect in synergy to anergy in oral tolerance of Blomia tropicalis/ovalbumin murine co-immunization model. AB - Oral tolerance is an important approach in allergic diseases and murine model can provide useful information to improve its understanding and therapeutic measures. To address the influence of non-related allergen sensitization in immunized mice with the mite Blomia tropicalis (Bt) or ovalbumin (OVA) or with both Bt/OVA allergens. Furthermore, we sought to verify oral tolerance effect in the Bt/OVA co-immunization model. Mice sensitized with Bt and then exposed to OVA developed an enhanced IgE response to both allergens; contrariwise, this effect was not observed when OVA-sensitization was prior to Bt-sensitization. Co-injection of Bt and OVA led to a dominant IgE response towards OVA over Bt, which was not observed when co-immunization was performed with a 240-fold less amount of OVA. Induction of oral tolerance with OVA, prior to co-immunization, suppressed IgE response to both allergens, probably as a consequence of the increased levels of IFN-gamma found in these animals. The results evidenced that, depending on allergenic potential, new allergen exposure may exert an adjuvant effect to the first allergen used in the sensitization. The bystander suppression to non related allergens through oral tolerance should be a useful mechanism to control sensitization to new allergens. PMID- 15821893 TI - Distribution and clinical aspects of primary immunodeficiencies in a Taiwan pediatric tertiary hospital during a 20-year period. AB - Recent advances in immunologic techniques have lead to increased recognition of primary immunodeficiencies. A review of patients with suspected immunodeficiencies in a Taiwan tertiary hospital from January 1985 to October 2004 and molecular/genetic analyses done on some patients were investigated. Of the 403 patients selected based on the International Classification of Disease, Ninth Revision, 37 patients with PID (8 females and 29 males) were identified: 17 (46%) with antibody production deficiencies, nine (24%) with defective phagocyte function, four (11%) with combined B and T cell immunodeficiencies, seven (19%) with T cell deficiencies, but none with primary complement deficiencies. Those with secondary immunodeficiencies were excluded from the study. Recurrent sinopulmonary infections (62%) were the most common clinical manifestation, followed by sepsis (57%), severe skin infection (40%), splenomegaly/hepatomegaly (27%), central nervous system dysfunction (22%), chronic diarrhea (22%), and failure to thrive (19%). Seven (19%) patients died, five of infections, one of disseminated intravascular coagulopathy and one of hepatocellular carcinoma. Six novel mutations were found from 11 agreed patients. This is the first report on primary immunodeficiencies in Taiwan covering a 20-year period. PMID- 15821894 TI - Benefits of postoperative shed blood reinfusion in patients undergoing unilateral total knee replacement. AB - INTRODUCTION: In patients undergoing total knee replacement (TKR), most of the measured blood loss occurs during the postoperative period, and 30-50% of these patients receive allogeneic blood transfusion (ABT). For this reason, the salvage and return of unwashed filtered shed blood (USB) from postoperative drainage may represent an alternative to ABT in these patients. We have, therefore, evaluated the clinical utility of USB return in TKR patients, with a special focus on patients with mild anaemia. MATERIALS AND METHODS: Data from 200 TKR patients (group 2) receiving USB within the first 6 postoperative hours (ConstaVac CBC II, Sryker) were prospectively collected. A retrospective series of 100 TRK patients served as the control group (group 1). RESULTS: USB return was possible in 162 patients who received a mean of 0.98+/-0.4 U/pte, without any clinically relevant incident. Return of USB decreased both the percentage of patients with ABT (48% vs 11%, for groups 1 and 2, respectively; p < 0.01) and the ABT units/patient index (1.31+/-1.27 vs 0.29+/-0.87 units/patient, respectively; p < 0.01). A transfusion protocol was not established, but there was no difference between groups with respect to either perioperative Hb levels or overall transfusion index, indicating that the transfusion criteria were uniform. However, for the subgroups of patients who needed ABT, the preoperative Hb level was 1 g/dL lower in those receiving USB (13.4+/-1.4 vs 12.4+/-1.2 g/dL; p < 0.05). There was no difference in the postoperative complication rate, and patients in group 2 recovered the ability to walk 1 day earlier, and their hospital stay was 3 days shorter than in group 1 (p < 0.01). CONCLUSIONS: Return of USB after TKR seems to shorten the hospital stay and effectively reduce postoperative requirements for ABT, especially in patients with preoperative Hb > or = 13 g/dL. For patients with preoperative Hb < 13 g/dL, although the return of USB also decreased the requirements for ABT, a further reduction will probably be obtained with the addition of another blood-saving method. PMID- 15821895 TI - Functional outcome in patients with zone V flexor tendon injuries. AB - INTRODUCTION: Zone V flexor tendon injuries may involve major nerves and arteries as well as the wrist and finger flexors. Although these injuries are not infrequent, few studies have reported functional outcomes. The purpose of this study was to evaluate the functional outcome in patients with flexor tendon repairs in zone V. MATERIALS AND METHODS: Eighteen patients with repaired zone V flexor tendon injuries were followed up for an average of 20 months. The postoperative rehabilitation program consisted of a combined regime of modified Kleinert and modified Duran techniques. Outcome parameters were hand function according to the Buck-Gramcko assessment system, grip and key pinch strength values, and return to work status. RESULTS: Functional results were excellent in 92.8% of the digits, good in 1.4%, and poor in 5.8%. Grip strength recovered to an average of 77% and pinch strength to 74% of the uninjured hand. Two tendon ruptures occurred in a patient, and tenolysis was required in 3 patients. Of 15 patients who were employed at the time of injury, 13 returned to their original occupations. CONCLUSION: Satisfactory functional results can be obtained when proper surgical technique is coupled with careful postoperative management in patients with zone V flexor tendon injuries. PMID- 15821896 TI - Diagnostic values of positron emission tomography versus triple-phase bone scan in hip arthroplasty loosening. AB - INTRODUCTION: The most frequent complications of total hip arthroplasty are septic and aseptic wear-induced loosening. A reliable differentiation between septic and aseptic loosening with current diagnostic tools is not possible. Therefore, we examined the diagnostic valency of positron emission tomography (PET) with (18)F-fluorodeoxyglucose (FDG) in cases of septic or aseptic hip arthroplasty loosening compared with conventional triple-phase bone scan (TPBS). MATERIALS AND METHODS: Fifty patients with 70 total hip replacements (symptomatic n=50, asymptomatic n=20) were examined by means of FDG-PET and TPBS to detect septic and aseptic loosening and differentiate between the two. A differentiated algorithm subdivided into categories I-V was developed for FDG-PET. Additionally, standardized uptake values (SUV) were calculated from the lesion with the highest FDG uptake. Interpretations of the TPBS were done according to the criteria described by Wilson. The final diagnosis was based on operative findings including microbiological and histological examinations (n=50), while the remaining asymptomatic arthroplasties (n=20) were integrated into a clinical follow-up (> or =9 months). RESULTS: Sensitivity/specificity of FDG-PET was 91%/92% (accuracy 91%) compared with 78%/70% (accuracy 74%) for TPBS. A high correlation could be proved between FDG-PET investigation and operative histopathological findings (r(Spear)> or =0.9). No significant differences were found regarding cemented and uncemented implanted hip arthroplasties (p> or =0.05). Calculation of the SUV turned out to be inappropriate as a sole criterion for image interpretation. CONCLUSION: FDG-PET is a promising, highly accurate examination method to detect polyethylene and metal wear-induced chronic inflammation followed by periprosthetic osteolysis. In addition, FDG-PET has a significantly higher sensitivity and specificity than TPBS for differentiating between aseptic loosening and infection. PMID- 15821897 TI - Painful non-union of the fibula in football player following fracture of the leg. AB - We report the case of a young football player operated on for a middle shaft fracture of the right leg with a Kuntscher nail. After 8 months, when he started to play football again, he suffered from a painful pseudoarthrosis of the region at the transition from the median to the lower third of the fibula. Because of the severity of the pain, and due to the lack of a well-defined and stated surgical option, a subtraction osteotomy was performed in order to remove the cause of the ache. The patient was revisited 3 and 6 months later, and showed complete disappearance of the pain and a full return to sport activities. In light of the good result obtained, we consider the osteotomy operation a reliable and easy-to-perform method in treating non-unions of the fibula. PMID- 15821898 TI - The crisscross injury mechanism in forearm injuries. AB - INTRODUCTION: A new mechanism of injury of the forearm bones, crisscross injury, is described. It is more common than the Essex-Lopresti fracture dislocation. The old concept of isolated injury of one side of the radioulnar joint may be challenged. It often occurs in Mason type II fracture dislocation of the radial head or dislocation of radioulnar joints. MATERIALS AND METHODS: The first part was a cadaveric study of the crisscross injury of forearms. The second part was a clinical study of the crisscross injury in some cases of Mason type II fracture radial head and double dislocation of the radioulnar joint. RESULTS: The cadaveric study confirmed a stable crisscross fracture dislocation injury with intact interosseous membrane. The clinical study echoed the presence of this injury by imaging techniques. CONCLUSION: The crisscross injury mechanism explains the mirror pathogenesis of the traumatic fracture dislocation of the distal and proximal radioulnar joints with intact shaft of the radius and ulna. Co-existing subluxation or dislocation of the other radioulnar articulation must not be overlooked in cases of fracture dislocation of one radioulnar joint. Two types of crisscross injury of forearm bones are proposed. PMID- 15821899 TI - The effect of low intensity ultrasound and bioabsorbable self-reinforced poly-L lactide screw fixation on bone in lateral malleolar fractures. AB - INTRODUCTION: There are no previous reports of low intensity pulsed ultrasound therapy in connection with bioabsorbable fracture fixation. In this randomised, prospective, double-blind and placebo-controlled study, the effects of ultrasound on bone mineral density and bone healing were examined in lateral malleolar fractures fixed with a bioabsorbable self-reinforced poly-L-lactide screw (SR PLLA). MATERIALS AND METHODS: Thirty adult patients with SR-PLLA screw-fixed lateral malleolar fracture underwent ultrasound therapy 20 min daily for 6 weeks. Half of the patients were provided randomly with a sham ultrasound device. Bone mineral density and bone healing were assessed by dual-energy X-ray absorptiometry (DXA) and by radiographs. RESULTS: Bone mineral density of the fractured lateral malleolus tended to increase slightly during 12 weeks of follow up. The increase was symmetrical and statistically non-significant between the ultrasound and non-ultrasound group. All the fractures healed uneventfully. The biocompatibility of the bioabsorbable SR-PLLA fixation device and low intensity pulsed ultrasound was good. Despite the slight tendency for more frequent callus formation in the ultrasound group, no statistically significant effect of low intensity pulsed ultrasound on lateral malleolar fracture healing could be observed. CONCLUSION: It was not possible to observe any statistically significant effect of low intensity pulsed ultrasound on lateral malleolar fracture healing in this study. Further studies are needed to examine the role of ultrasound therapy in the healing of fractures treated with bioabsorbable fixation devices. PMID- 15821900 TI - [Infections after organ transplantation]. AB - Early postoperative infections after transplantation vary according to the transplanted organ. During the subsequent course opportunistic infections such as cytomegalovirus reactivation, Pneumocystis jiroveci pneumonia, invasive pneumococcal infection and mould infections predominate. Reactivated tuberculous infection appears to become more prevalent. Some of the opportunistic infections are preventable by chemoprophylaxis; others can be managed very effectively by monitoring and early preemptive therapy. Physicians caring for patients after organ transplantation need to early consider in the differential diagnosis rare pathogens which are often overlooked with standard diagnostic procedures. PMID- 15821903 TI - [Sub-specialization within the field of radiology]. AB - Medical knowledge and skills are accumulating at an ever increasing speed. Therefore, it becomes more and more difficult for a single one to have a complete overview of a whole discipline. This general observation also applies for radiology. Radiology has to cooperate with almost all medical disciplines and has to provide competent experts. It is therefore mandatory to support differentiation within the field of radiology and at the same time to save coherence and future developments. PMID- 15821902 TI - Studies of variations of the cyclin-dependent kinase inhibitor 1C and the cyclin dependent kinase 4 genes in relation to type 2 diabetes mellitus and related quantitative traits. AB - CDK4 is involved in the regulation of body weight, pancreatic beta-cell proliferation, insulin responsiveness, and diabetes pathogenesis. CDK4 activity is inhibited by CDKN1C, which is regulated by insulin. In addition, CDKN1C plays an important role in beta-cell proliferation and is involved in the pathogenesis of the Beckwith-Wiedemann syndrome, a disorder characterized by neonatal hyperinsulinaemic hypoglycaemia and pre- and post-natal overgrowth. The aim of this study was to investigate if variations in the proximal promoter and the coding region of the CDKN1C and CDK4 genes are associated with type 2 diabetes or changes in related quantitative phenotypes among glucose-tolerant subjects. Mutation analyses of the two genes in 62 type 2 diabetic patients resulted in the discovery of seven variants of CDKN1C and two variants of CDK4. In a case-control study comprising 717 type 2 diabetic patients and 518 glucose-tolerant subjects the most frequent variants did not show any difference in allele frequencies between the type 2 diabetic patients and the control subjects. However, in two genotype-quantitative trait correlation studies involving 206 glucose-tolerant offspring of type 2 diabetic patients and 359 young, healthy subjects the CDKN1C del171APVA variant associated with increased birth weight (P=0.05 and P=0.05). Furthermore, the same variant tended to be associated with decreased basal glucose oxidation among 16 genotypically discordant dizygotic twins (P=0.03). In a genotype-quantitative trait study involving 500 middle-aged glucose-tolerant subjects the CDK4 IVS2-31G-->A variant was associated with an increased waist circumference (P=0.03) and waist-to-hip ratio (P=0.02) and altered fasting plasma glucose (P=0.03). However, these later findings could not be replicated in additional studies. In conclusion, variants in CDKN1C may contribute to the inter individual variation in birth weight. PMID- 15821904 TI - Monolayers of photosystem II on gold electrodes with enhanced sensor response- effect of porosity and protein layer arrangement. AB - Mass transport of the bulk of the analyte to the electrode and through the bioactive layer can be significantly improved by use of the nanoelectrode array and defined arrangement of protein film. This phenomenon has been studied by (i) atomic-force microscopy, (ii) electrochemical measurements of PSII activity, and (iii) digital simulations for an oriented monolayer of histidine-tagged photosystem II (PSII) immobilized on nitrilotriacetic acid (NTA)-modified gold electrodes. The output signal of the electrochemical biosensor is controlled by (i) mass transport from the bioactive layer to electrode and (ii) mass transport between the bulk of the analyte and the electrode. Mass transport through the bioactive layer was electrochemically studied for PSII self-assembled on gold screen-printed electrodes. A densely packed monolayer of PSII has a significant shielding effect toward the diffusion of redox mediator duroquinone (DQ). Mass transport to the planar electrode surface was improved by co-immobilization of bovine-serum albumin (BSA) as spacer biomolecule in the monolayer of PSII. Correlation between the electrochemical properties and surface arrangement of the resulting protein films was clearly observable and confirmed the improved mass transport properties of structured enzyme monolayers. On the basis of this observation, the application of a bottom-up approach for improvement of electrode performance was proposed and digitally simulated for an infinite array of electrodes ranging in diameter from 50 nm to 5 microm. The nanoelectrode array, with the optimum time window selected for measurements, enables enhancement of mass transport between the bulk of the analyte and the macroelectrode by a factor of up to 50 in comparison with "classical" planar electrodes. Use of a time window enables minimization of crosstalk between individual electrodes in the array. The measurements require methods which suppress the double-layer capacity. PMID- 15821901 TI - Human defensins. AB - Antimicrobial peptides are small, cationic, amphiphilic peptides of 12-50 amino acids with microbicidal activity against both bacteria and fungi. The eukaryotic antimicrobial peptides may be divided into four distinct groups according to their structural features: cysteine-free alpha-helices, extended cysteine-free alpha-helices with a predominance of one or two amino acids, loop structures with one intramolecular disulfide bond, and beta-sheet structures which are stabilised by two or three intramolecular disulfide bonds. Mammalian defensins are part of the last-mentioned group. The mammalian defensins can be subdivided into three main classes according to their structural differences: the alpha-defensins, beta defensins and the recently described theta-defensins. Mammalian alpha-defensins are predominantly found in neutrophils and in small intestinal Paneth cells, whereas mammalian beta-defensins have been isolated from both leukocytes and epithelial cells. Recently, two novel human beta-defensins, human beta-defensin-3 (HBD-3), and human beta-defensin-4 (HBD-4) have been discovered. Similar to HBD-1 and HBD-2, HBD-3 has microbicidal activity towards the Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli) and the yeasts Candida albicans and Malassezia furfur. In addition, HBD-3 kills Gram-positive bacteria such as Streptococcus pyogenes or Staphylococcus aureus, including multi-resistant S. aureus strains, and even vancomycin-resistant Enterococcus faecium. In contrast to HBD-1 and HBD-2, significant expression of HBD-3 has been demonstrated in non epithelial tissues, such as leukocytes, heart and skeletal muscle. HBD-4 is expressed in certain epithelia and in neutrophils. Its bactericidal activity against P. aeruginosa is stronger than that of the other known beta-defensins. Here we present an overview of human antimicrobial peptides with some emphasis on their antifungal properties. PMID- 15821905 TI - Detoxification of mercury in the environment. AB - In this study, a "green chemistry" approach was developed as an option for remediation of toxic mercury in the environment. Twenty mercury compounds were treated with an environmentally friendly agent cyclodextrin to produce stable non toxic mercury in soil and water. The binding efficiency was determined using high performance liquid chromatography with diode-array detection. The stability of the cyclodextrin mercury complexes toward environmental microorganisms in water was estimated under OECD guidelines using gas chromatography-mass spectrometry. The toxicity of the cyclodextrin mercury compounds to terrestrial organisms was investigated by use of internationally recognized toxicity methods using mercuric acetate as a model contaminant. Key process conditions, for example pH, temperature, and amount of detoxifying agent were investigated and found to have significant effects on the toxicity of mercury. It was found that organic and inorganic mercury pollutants could be mineralized in the environment with cyclodextrins. The bound mercury compounds resisted biodegradation and were found to be non-toxic to environmental microorganisms under laboratory conditions. PMID- 15821906 TI - Investigation of Helicobacter pylori colonization in laryngeal neoplasia. AB - Helicobacter pylori has been investigated in several other organ systems and localizations such as the oral cavity, but has not been investigated extensively in squamous cell carcinoma of the larynx, a region that could be directly exposed to the bacterium by the oral route or gastro-esophageal reflux. Only serological studies are available regarding the relation between H. pylori and laryngeal cancer, yielding conflicting results. To our knowledge, there is no study investigating the presence of H. pylori in laryngeal squamous cell carcinoma tissue. The purpose of this study was to investigate the presence of H. pylori in laryngeal squamous cell carcinoma tissue and to investigate the possible role of this organism in the etiopathogenesis of laryngeal cancer. Specimens from 50 patients with laryngeal cancer who underwent total or partial laryngectomy between March 1999 and December 2002 were examined by histopathological and immunohistochemical methods to detect H. pylori. The presence of H. pylori was also investigated histopathologically in 50 benign laryngeal biopsy specimens. In our study, we demonstrated that H. pylori was not present in laryngeal squamous cell carcinoma tissue or in the benign lesions. We could not find any evidence indicating that H. pylori played a role at the tissue level in the pathogenesis of laryngeal carcinoma. PMID- 15821907 TI - Hoarseness as an unusual initial presentation of aortic dissection. AB - Aortic dissection is a life-threatening medical emergency. While an abrupt, tearing pain in the chest or back is present in more than 90% of the patients, diagnosis of aortic dissection has been shown to be particularly difficult when such symptoms are not present. In this report we describe a 36-year-old man presenting with a 10-day history of new onset of hoarseness associated with several transient headache episodes. The possibility of aortic dissection was overlooked at the initial presentation, and unilateral vocal cord palsy due to a cerebrovascular accident was the initial diagnosis. The patient's abnormal chest radiograph led to a re-diagnosis, and a Standford type-B aortic dissection was confirmed using thoracic computed tomography. The unusual presentation of aortic dissection is emphasized, and its management is discussed. PMID- 15821908 TI - A safe and cost-effective method of removal of obstructed pharyngeal foreign body in the accident and emergency department. AB - Foreign body obstruction at the level of the cricopharynx is not an uncommon presentation in the accident and emergency department. If this is visualised with a fibro-optic endoscope, it can be safely removed in the emergency department with the help of short acting inhalational anaesthesitic agents such as sevoflurane. This method is safe and effective and also saves valuable operating theatre resources. PMID- 15821909 TI - Videofluoroscopic evaluation of the swallowing function after supracricoid laryngectomy. AB - This study was designed to evaluate the swallowing function in patients with supracricoid laryngectomy (SCL) compared to normal subjects and to search for the factors affecting postoperative aspiration. Ten patients who underwent SCL with cricohyoidopexy (CHP) for primary laryngeal squamous cell carcinoma were included in the study. The control group consisted of 13 normal adult volunteer men with similar ages. The swallowing act of the subjects was evaluated by using videofluoroscopy (VFS) and videolaryngostroboscopy (VLS). The movements of the larynx were measured with regard to the hyoid bone, mandible and vertebral spine. The patients with SCL-CHP, except for two who had slight aspiration, had effective and near normal swallowing regarding the measurements of the movements of the hyoid bone. They could tolerate a near-normal oral diet. We have observed that the preventive precautions for aspiration are preserving the superior laryngeal nerves, suturing and positioning the cricoarytenoid unit as anterosuperiorly as possible, early decannulation and early onset of swallowing rehabilitation; the risk factors for aspiration are advanced stage of cancer, postoperative radiation and shortening of bolus transit time. VFS is useful for the patients with postoperative aspiration, because it is the definitive technique for anatomical and physiological evaluation of swallowing. We consider that the parameters of VLS and VFS, such as tongue base-arytenoid contact, presence of bolus splitting, pseudoepiglottis function, maximal opening of the pharyngoeosophageal sphincter and total movement of hyoid bone are important criteria to evaluate swallowing. PMID- 15821910 TI - Patient discomfort in polysomnography with esophageal pressure measurements. AB - The reference method for measuring respiratory effort and for differentiating between obstructive and central apneas in the diagnosis of sleep-related breathing disorders is overnight monitoring of esophageal pressure. Despite this being the reference method, it is not widely used because it is considered invasive and uncomfortable for the patients. The aim of this study was to assess patient discomfort and insertion difficulty when using an esophageal catheter during polysomnography. We have performed a prospective questionnaire-based clinical study in 799 consecutive patients where polysomnography with an esophageal catheter was routinely performed in the diagnosis of sleep-related breathing disorders. The main outcome measures were the catheter-related discomfort experienced by the patient and difficulty of catheter insertion reported by the sleep technician. Ninety-six percent of the patients accepted the insertion of the catheter, and most of the patients considered it acceptable to sleep with the catheter. Correspondingly, in most of the patients, the catheter was easily inserted, and there were difficulties in only a few patients. Specifically, no complications or side effects were reported when using the catheter. To optimize the diagnosis of sleep-related breathing disorders, an esophageal sensor catheter can be used during polysomnography, without causing major patient discomfort. PMID- 15821911 TI - High yield of long-chain polyunsaturated fatty acids by labyrinthulids on soybean lecithin-dispersed agar medium. AB - The aim of the present research was to provide an effective long-chain polyunsaturated fatty acid (LCPUFA) production by labyrinthulids using soybean lecithin (SBL). Use of SBL-dispersed agar medium resulted in higher LCPUFA production than soybean oil. Among the components of SBL, phosphatidylinositol (PI) and triacylglycerol (TG) were revealed to be essential factors for high growth of labyrinthulids. PI was effective for surface growth, and TG was effective for three-dimensional growth. The presence of some elements like carotenoids in SBL and the smaller droplet size of dispersed SBL were also attributed to be factors for the higher LCPUFA productivity on SBL medium. LCPUFA productivity and the volume of the oval form of the cells increased with increasing SBL concentration up to 40 g/l. Under optimum conditions, LCPUFA production of the L25 strain, isolated from Ogasawara Island in Japan, reached 2.91 g/l after 21 days. PMID- 15821912 TI - Cloning and expression of a gene for an alpha-glucosidase from Saccharomycopsis fibuligera homologous to family GH31 of yeast glucoamylases. AB - Cloning of cDNA encoding an alpha-glucosidase from the dimorphous yeast Saccharomycopsis fibuligera and characterization of the gene product were performed. The cDNA of the putative alpha-glucosidase gene consists of 2,886 bp, which includes an open reading frame encoding a 19 amino acid signal peptide at the N-terminal end and a 944 amino acid mature protein with a predicted molecular mass of 105.4 kDa and pI value of 4.52. The deduced amino acid sequence shows a high degree of identity (70%) with two yeast glucoamylases, namely, the extracellular glucoamylase Gam from Schwanniomyces occidentalis and the cell surface glucoamylase Gca from Candida albicans. The recombinant product, synthesized in Saccharomyces cerevisiae, is localized on the cell surface and hydrolyses maltooligosaccharides exclusively without the ability to digest soluble starch, which is consistent with the specificity characteristic of alpha glucosidase, EC. 3.2.1.20. PMID- 15821913 TI - Gene expression analysis of Escherichia coli grown in miniaturized bioreactor platforms for high-throughput analysis of growth and genomic data. AB - Combining high-throughput growth physiology and global gene expression data analysis is of significant value for integrating metabolism and genomics. We compared global gene expression using 500 ng of total RNA from Escherichia coli cultures grown in rich or defined minimal media in a miniaturized 50-microl bioreactor. The microbioreactor was fabricated out of poly(dimethylsiloxane) (PDMS) and glass and equipped to provide on-line, optical measurements. cDNA labeling for microarray hybridizations was performed with the GeniconRLS system. From these experiments, we found that the expression of 232 genes increased significantly in cells grown in minimum medium, including genes involved in amino acid biosynthesis and central metabolism. The expression of 275 genes was significantly elevated in cells grown in rich medium, including genes involved in the translational and motility apparatuses. In general, these changes in gene expression levels were similar to those observed in 1,000-fold larger cultures. The increasing rate at which complete genomic sequences of microorganisms are becoming available offers an unprecedented opportunity for investigating these organisms. Our results from microscale cultures using just 500 ng of total RNA indicate that high-throughput integration of growth physiology and genomics will be possible with novel biochemical platforms and improved detection technologies. PMID- 15821914 TI - Amphotericin B. AB - Invasive fungal infections are a major cause of morbidity and mortality in immunodeficient individuals (such as AIDS patients) and in transplant recipients or tumor patients undergoing immunosuppressive chemotherapy. Amphotericin B is one of the oldest, yet most efficient antimycotic agents. However, its usefulness is limited due to dose-dependent side-effects, notably nephrotoxicity. In order to improve its safety margin, new pharmaceutical formulations of amphotericin B have been designed especially to reduce its detrimental effects on the kidneys. Since the 1980s, a wide variety of new amphotericin B formulations have been brought forward for clinical testing, many of which were approved and reached market value in the 1990s. This review describes and discusses the molecular genetics, pharmacological, toxicological, and clinical aspects of amphotericin B itself and many of its innovative formulations. PMID- 15821915 TI - Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome. AB - A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coli-Streptomyces artificial chromosome (ESAC) and screened for the presence of dbv genes known to be involved in the biosynthesis of the glycopeptide A40926. dbv genes were cloned as two large, partially overlapping, fragments and transferred into the host Streptomyces lividans, thus generating strains S. lividansColon, two colonsNmESAC50 and S. lividansColon, two colonsNmESAC57. The heterologous expression of Nonomuraea genes in S. lividans was successfully demonstrated by using combined RT-PCR and proteomic approaches. MALDI-TOF analysis revealed that a Nonomuraea ABC transporter is expressed as two isoforms in S. lividans. Moreover, its expression may not require a Nonomuraea positive regulator at all, as it is present at similar levels in both clones even though S. lividansColon, two colonsNmESAC57 lacks regulatory genes. Considered together, these results show that S. lividans expresses Nonomuraea genes from their own promoters and support the idea that S. lividans can be a good host for genetic analysis of Nonomuraea. PMID- 15821916 TI - Expression and characterization of a thermostable serine protease (TfpA) from Thermomonospora fusca YX in Pichia pastoris. AB - A serine protease produced by Thermomonospora fusca YX (TfpA) is heat-stable (up to 85 degrees C) and has a broad pH activity range and strong resistance to detergents. The objective of this study was to determine if the methylotropic yeast Pichia pastoris could express TfpA extracellularly. A 1.0-kb DNA fragment (tfpA) encoding the pro-peptide and mature protein of TfpA was cloned into expression vectors pPICZalphaA (inducible) and pGAPZalphaA (constitutive) and introduced into P. pastoris by electroporation. Expression of r-TfpA was greater in the inducible system than in the constitutive one, producing 135 U ml(-1) medium supernatant 6 days after methanol induction. The r-TfpA was not glycosylated (21.7 kDa), and had pH and temperature optima of 8.5 and 80 degrees C, respectively, using azocasein as a substrate. In conclusion, P. pastoris can be used as a host to produce extracellular r-TfpA, and expression efficiency may be improved by optimizing fermentation conditions and modifying factors related to protein expression and stability. PMID- 15821917 TI - [Epidemic keratoconjunctivitis. Detecting adenoviruses]. AB - OBJECTIVE: The validity of the 15-min adenovirus assay SAS Adenotest was evaluated compared with virus detection by polymerase chain reaction (PCR) from conjunctival swabs. METHODS: In 75 patients with assumed epidemic keratoconjunctivitis, adenovirus detection from conjunctival swabs was performed by the immunochromatographic assay SAS Adenotest and PCR. RESULTS: In 25 patients adenovirus was detected by PCR, 18 of whom were detected by the SAS Adenotest and 7 of whom were not. No false positive results occurred. Sensitivity was 72% and specificity was 100%. CONCLUSIONS: The results indicate that rapid adenovirus detection with the SAS Adenotest is a useful tool in early epidemic keratoconjunctivitis. An additional PCR should be performed when clinical symptoms persist for 5 days or more. PMID- 15821918 TI - [Unilateral temporary vision loss in an otherwise healthy elderly patient]. PMID- 15821919 TI - [Diplopia after delivery of a child]. PMID- 15821920 TI - [Autologous serum versus hyaluronic acid eye drops for the treatment of corneal erosions after vitrectomy in diabetic patients. A prospective randomized study]. AB - PURPOSE: The purpose of this study was to evaluate whether treatment with autologous serum exhibits better epitheliotropic properties than hyaluronic acid in patients with reduced wound healing (diabetics). PATIENTS AND METHOD: This prospective randomized study included all diabetic patients undergoing pars plana vitrectomy with abrasion of the corneal epithelium. In addition to the usual topical therapy one group was treated with hourly application of autologous serum, the other group with hyaluronic acid drops. We evaluated the time needed for closure of the corneal epithelium as well as other parameters such as patient's age, sex, duration of surgery, history of diabetes, and the current HbA1C. RESULTS: There were 23 patients enrolled in the study. The epithelium healed a mean of 2 days earlier in the autologous serum group. Based on the statistically significant difference in favor of autologous serum, the study was terminated after treatment of 23 patients. CONCLUSIONS: Autologous serum eye drops lead to earlier closure of the corneal epithelium after intraoperative abrasion in diabetic patients when compared with artificial tears. PMID- 15821922 TI - Abstracts of the 7th International Congress on Spinal Surgery. Antalya, Turkey, 14-17 April 2005. PMID- 15821921 TI - Incidental durotomy in lumbar spine surgery: incidence and management. AB - There is increasing awareness of the need to inform patients of common complications that occur during surgical procedures. During lumbar spine surgery, incidental tear of the dural sac and subsequent cerebrospinal fluid leak is possibly the most frequently occurring complication. There is no consensus in the literature about the rate of dural tears in spine surgery. We have undertaken this study to evaluate the incidence of dural tears among spine surgeons in the United Kingdom for commonly performed spinal procedures. Prospective data was gathered for 1,549 cases across 14 institutions in the United Kingdom. The results give us a baseline rate for the incidence of dural tears. The rate was 3.5% for primary discectomy, 8.5% for spinal stenosis surgery and 13.2% for revision discectomy. There was a wide variation in the actual and estimated rates of dural tears among the spine surgeons. The results confirm that prospective data collection by spine surgeons is the most efficient and accurate way to assess complication rates for spinal surgery. PMID- 15821924 TI - Detection of human beta defensin-1 and -2 by RT-competitive multiplex PCR. AB - Although, the human epithelium is constantly challenged by a broad spectrum of microorganisms, invasive infections are rather rare. Recent findings suggest the expression of antimicrobial peptides by skin cells in order to provide an innate defensive barrier. In particular, peptides of the beta-defensin family offer antimicrobial activity against different pathogens including bacteria and fungi. Within this peptide family, hBD-1 is rather constitutively expressed while hBD-2 and hBD-3 expression depends on environmental conditions. The present paper introduces RT-competitive multiplex PCR as a precise tool to detect hBD-1 and hBD 2 expression on the transcriptional level. The method makes use of co amplification of synthetic competitors along with referring wildtype targets. Competitor- and wildtype-derived products differ in size allowing signal discrimination using agarose gel electrophoresis. Regulation of gene transcripts is evaluated by comparison of competitor and corresponding wildtype signals. It was found that primary human keratinocytes stimulated with Escherichia coli cells for 8 h offered an upregulation of hBD-2 to about 2,000 fold, while hBD-1 was only marginally regulated. RT-competitive multiplex PCR is a simple and accurate method that enables new insights into defensin regulation under physiological and pathophysiological conditions. PMID- 15821925 TI - Low-dose ionizing radiation: induction of differential intracellular signalling possibly affecting intercellular communication. AB - Given the complexity of the carcinogenic process and the lack of any mechanistic understanding of how ionizing radiation at low-level exposures affects the multistage, multimechanism processes of carcinogenesis, it is imperative that concepts and paradigms be reexamined when extrapolating from high dose to low dose. Any health effect directly linked to low-dose radiation exposure must have molecular/biochemical and biological bases. On the other hand, demonstrating some molecular/biochemical or cellular effect, using surrogate systems for the human being, does not necessarily imply a corresponding health effect. Given the general acceptance of an extrapolated LNT model, our current understanding of carcinogenesis cries out for a resolution of a real problem. How can a low-level acute, or even a chronic, exposure of ionizing radiation bring about all the different mechanisms (mutagenic, cytotoxic, and epigenetic) and genotypic/phenotypic changes needed to convert normal cells to an invasive, malignant cell, given all the protective, repair, and suppressive systems known to exist in the human body? Until recently, the prevailing paradigm that ionizing radiation brings about cancer primarily by DNA damage and its conversion to gene and chromosomal mutations, drove our interpretation of radiation carcinogenesis. Today, our knowledge includes the facts both that epigenetic events play a major role in carcinogenesis and that low-dose radiation can also induce epigenetic events in and between cells in tissues. This challenges any simple extrapolation of the LNT model. Although a recent delineation of "hallmarks" of the cancer process has helped to focus on how ionizing radiation might contribute to the induction of cancers, several other hallmarks, previously ignored--namely, the stem cells in tissues as targets for carcinogenesis and the role of cell-cell communication processes in modulating the radiation effects on the target cell- must be considered, particularly for the adaptive response, bystander effects, and genomic instability phenomena. PMID- 15821926 TI - Effects of a selective glucocorticoid receptor agonist on experimental keratoplasty. AB - BACKGROUND: New insights into the molecular mechanisms of corticosteroid-mediated actions have revealed new substances, such as selective glucocorticoid receptor agonists (SEGRA), for the treatment of inflammatory diseases. We set out to evaluate the effect of a SEGRA compound following topical application on the course of experimental orthotopic corneal grafts. METHODS: A total of 42 female Lewis rats received 3.5-mm MHC I/II-incompatible corneal grafts from DA donors. Recipients were randomly assigned to receive either no therapy, 0.25% cyclodextrin-encapsulated SEGRA compound in a new microemulsion formulation or carrier system only. All treatments started on the day of surgery and were given five times daily for 35 days. Grafts were graded every day and a rejection score was generated based on cornea clarity and edema. In addition, intragraft mRNA expression of CD3, IFN-gamma, TNF-alpha, IL-10 and IL-4 was analyzed using real time RT-PCR analysis at day 7 after transplantation before rejection occurred in additional control animals. RESULTS: Topical application of a SEGRA compound was highly effective in prolonging the mean survival time of corneal grafts (42.2+/ 4.0 days) compared with untreated controls (11.7+/-1.2 days, p=0.00003) or animals that received the vehicle only (15.0+/-1.5 days, p=0.114). In addition, real-time RT-PCR analysis of SEGRA-treated grafts revealed lower mRNA expression of intragraft cytokines; the difference was significant for IL-4 (p<0.05). CONCLUSIONS: Our results indicate that topical application of a SEGRA compound significantly prolongs corneal graft survival in an experimental keratoplasty model. It further suggests that SEGRA can be a potentially useful drug to suppress the immune response. PMID- 15821927 TI - Iron accumulation does not parallel the high expression level of ferritin in transgenic rice seeds. AB - To answer the question whether iron accumulation in transgenic rice seeds depends on the expression level of exogenous soybean ferritin, we generated two kinds of ferritin hyper-expressing rice lines by introducing soybean ferritin SoyferH-1 gene under the control of the rice seed storage glutelin gene promoter, GluB-1 and the rice seed storage globulin gene promoter, Glb-1, (GluB-1/SoyferH-1 and Glb-1/SoyferH-1, DF lines), and by introducing the SoyferH-1 gene under the control of Glb-1 promoter alone (Glb-1/SoyferH-1, OF lines). Ferritin expression was restricted to the endosperm in both lines and protein levels determined by western blot analysis were up to 13-fold higher than in a construct previously reported FK22 (GluB-1/SoyferH-1, in genetically Kitaake background); [corrected] however, the maximum iron concentrations in seeds of both of the new lines were only about 30% higher than FK22. The maximum iron concentration in the OF and DF lines was about threefold higher than in the non-transformant. The mean Fe concentration in leaves of ferritin over-expressing lines decreased to less than half of the non-transformant while that the plant biomasses and seed yields of the ferritin-transformed lines were not significantly different from those of the non-transformant, suggesting that accumulation of Fe in seeds of hyper-expression ferritin rice did not always depend on the expression level of exogenous ferritin but may have been limited by Fe uptake and transport. No obvious differences were observed for other divalent-metal concentrations (Ca, Cd, Cu, Mg, Mn and Zn) in the seeds among all experimental lines and non-transformant. PMID- 15821928 TI - Analysis of trefoil factor family protein 1 (TFF1, pS2) expression in chronic cholecystitis and gallbladder carcinoma. AB - Trefoil factor family protein 1 (TFF1, pS2) interacts with mucins to protect gastrointestinal epithelium against injury and contributes to mucosal repair by promoting epithelial cell migration and restitution. Moreover, TFF1 has antiproliferative and anti-apoptotic effects and promotes cell scattering and invasion. We investigated TFF1 expression in healthy and inflamed non-neoplastic gallbladder mucosa as well as in gallbladder carcinomas (n=57) and corresponding metastases (n=18), using a tissue microarray technique. TFF1 immunoreactivity was absent in healthy mucosa, focally observed in epithelium with inflammatory changes and present in 35% of primary and 24% of metastatic cancer tissues. Immunoreactivity significantly decreased with increasing tumour stage (P=0.009) and increasing tumour grade (P=0.001). Patients with TFF1 positive tumours showed a more favourable outcome compared to patients with TFF1 negative tumours in univariate analysis (P=0.006). However, multivariate analysis proved resection status and tumour grade as the only independent prognostic factors. In conclusion, TFF1 is expressed in inflamed non-neoplastic gallbladder epithelium and in low stage and low grade gallbladder carcinomas. Thus, TFF1 may be the missing link between gallstones, chronic cholecystitis and gallbladder cancer. Further studies are needed to evaluate whether TFF1 immunostaining can be used as a diagnostic tool to identify patients with a more favourable outcome. PMID- 15821929 TI - Carcinosarcoma of the prostate: two cases with distinctive morphologic and immunohistochemical findings. AB - Carcinosarcomas (CS) of the prostate are very uncommon neoplasms defined by the admixture of malignant epithelial and mesenchymal components. We describe here two new examples of CS in two patients aged 66 and 77 years, the first without previous history of prostate adenocarcinoma and the second with a 5-year history of acinar type prostate adenocarcinoma. The diagnosis of CS was made on the cystoprostatectomy specimen in the first case and transurethral resection in the second case. Both biphasic tumours exhibited papillary areas of ductal differentiation and conventional adenocarcinoma in the epithelial component, as well as malignant fibrous histiocytoma and angiosarcomatous areas in the first case and solid, poorly differentiated epithelial areas with neuroendocrine features in the second case. Immunohistochemistry revealed over-expression of c erb B2 in the papillary epithelial component of both cases, whereas the solid undifferentiated epithelial areas in the second patient expressed c-kit, CD10 and synaptophysin, thus conforming a very undifferentiated cell population. The angiosarcomatous component of the first case expressed CD31 and CD10. The clinical course of the cases was divergent; the first patient is free of disease after radical surgery and adjuvant therapy and the other died 5 months after the diagnosis of CS, having already developed liver metastases. PMID- 15821930 TI - "Sugar" tumor of the pancreas: a rare entity that is diagnosable on preoperative fine-needle biopsies. AB - This study is the second to report a pancreatic "sugar" tumor (ST) case. This ST was incidentally discovered in a 31-year-old woman using computed tomography scan (CT scan) for work-up of a hepatic focal nodular hyperplasia. Both CT scan and endoluminal ultrasonography (EUS) features evoked a 15-mm large benign endocrine tumor. Pathological examination of EUS-guided fine-needle aspiration biopsies could not confirm this diagnosis. Laparoscopic corporeo-caudal pancreatectomy was performed. The tumor was intrapancreatic, well circumscribed, and organized in sheets of epithelioid cells. The tumor cells expressed HMB-45 but did not express epithelial or endocrine immunohistochemical markers. These histophenotypic features are those of an extra pulmonary ST, which belong to the PEComa family of tumors. Retrospective examination of preoperative biopsies evidenced the same histophenotypic features. This observation highlights that STs should be considered in preoperative differential diagnosis of pancreas tumors, since they may be treated by limited surgical resection. PMID- 15821931 TI - Biochemical characterisation of the proteins encoded by the DiGeorge critical region 6 (DGCR6) genes. AB - The DiGeorge critical region 6 (DGCR6) gene exists in two highly homologous copies (DGCR6 and DGCR6L) on chromosome 22q11 and is deleted in patients with velo-cardio-facial syndrome/DiGeorge syndrome (VCFS/DGS). The DGCR6 mRNA levels are increased in metastatic mammary tumour cells and regulate the expression of neighbouring genes at the 22q11 region. Newly developed monoclonal antibodies detected predominantly nuclear phosphoproteins of approximately 25 kDa, with low expression levels in the cytoplasm. Both proteins have half-lives of about 2.5 h. Exogenously expressed DGCR6 and DGCR6L migrated with slightly different mobility in SDS-gels in accordance with two immunoreactive bands observed for the endogenous proteins. DGCR6 is found at low levels in primary human fibroblasts or peripheral blood mononuclear cells, while tumour cells, B-cells transformed by EBV as well as activated normal human T cells, contain elevated levels of the proteins. The proteins are differentially expressed in mammalian tissues, with high protein levels in heart, liver and skeletal muscle. These observations are important as some patients with DGCR6 syndrome exhibit a T-cell deficiency and/or cardiac malformations. As the DGCR6 protein(s) influence gene expression in trans, we analysed the influence of DGCR6/DGCR6L on the Epstein-Barr virus encoded oncoproteins EBNA2 and EBNA3c in the activation of the viral LMP1 promoter, as well as LMP1-mediated activation of NFkB, but found no effect in either setting. PMID- 15821932 TI - Differential expression of the GABA transporters GAT-1 and GAT-3 in brains of rats, cats, monkeys and humans. AB - The homeostasis of GABA is critical to normal brain function. Extracellular levels of GABA are regulated mainly by plasmalemmal gamma-aminobutyric acid (GABA) transporters. Whereas the expression of GABA transporters has been extensively studied in rodents, validation of this data in other species, including humans, has been limited. As this information is crucial for our understanding of therapeutic options in human diseases such as epilepsy, we have compared, by immunocytochemistry, the distributions of the GABA transporters GAT 1 and GAT-3 in rats, cats, monkeys and humans. We demonstrate subtle differences between the results reported in the literature and our results, such as the predominance of GAT-1 labelling in neurons rather than astrocytes in the rat cortex. We note that the optimal localisation of GAT-1 in cats, monkeys and humans requires the use of an antibody against the human sequence carboxyl terminal region of GAT-1 rather than against the slightly different rat sequence. We demonstrate that GAT-3 is localised mainly to astrocytes in hindbrain and midbrain regions of rat brains. However, in species such as cats, monkeys and humans, additional strong immunolabelling of oligodendrocytes has also been observed. We suggest that differences in GAT distribution, especially the expression of GAT-3 by oligodendrocytes in humans, must be accommodated in extrapolating rodent models of GABA homeostasis to humans. PMID- 15821933 TI - Decreased EEG coherence between prefrontal electrodes: a correlate of high language proficiency? AB - To investigate the influence of proficiency level on the cortical organization of foreign language processing, two groups of German speaking students, differing only in their proficiency in English as a second language, were subjected to EEG coherence analysis during foreign and native language processing (news reports, alpha1 frequency band). In the group with minor experience with English, coherence increase was observed with all electrode combinations, with left hemisphere (LH) predominance. In the high proficiency group, coherence increase was limited to temporal electrodes over LH. In the latter group only, coherence between prefrontal electrodes was significantly lower during the language tasks than during the baseline task (silence, noisy screen). Both results were obtained with foreign as well as native language processing. We suggest that reduced EEG coherence in highly proficient foreign language speakers reflects a more efficient operating strategy not only for their second, but also for their native language. PMID- 15821934 TI - Timing the anticipatory recovery in smooth ocular pursuit during the transient disappearance of a visual target. AB - After the disappearance of a moving target and the loss of visual feedback regarding image motion, smooth pursuit eye velocity decays rapidly. If, however, there is an expectation the target will reappear further along its trajectory, there is a scaled recovery in eye velocity before target reappearance. The aim of this study was to examine whether the timing of the anticipatory recovery is influenced by the duration of transient target disappearance. We found that subjects (N=6) did not maintain eye velocity close to target velocity throughout the inter-stimulus interval (ISI). In general, after an initial reduction in eye velocity a significant increase was observed for most subjects before target reappearance, or a recovery that halted the decay. The timing of the recovery was not influenced by ISI even when this was predictable. There was, however, a significant effect of the initial visible ramp duration, indicating that the recovery was a consequence of the previous eye velocity trajectory and subsequent reacceleration. We suggest, therefore, that the recovery was timed to the moment of target disappearance rather than reappearance, and was the result of reactivation of a variable gain mechanism that acts on the visuomotor drive to ocular pursuit. PMID- 15821935 TI - Inspiratory and expiratory CT lung density in infants and young children. AB - BACKGROUND: There is a lack of information on normal inspiratory and expiratory CT lung density in infants. OBJECTIVE: To describe normal regional CT lung density at end inspiratory and end expiratory lung volumes in children ages 0--5 years. MATERIALS AND METHODS: Motionless HRCT images were obtained at 25 cm (inspiratory) and 0 cm (expiratory) water pressure at apical (top of arch) and basal (2 cm above diaphragm) levels in 16 sedated children (mean age 1.5 years) who underwent CT for reasons other than respiratory disease. Density was measured at anterior-posterior and medial-lateral locations at each level in each lung. The influence of level, location, and age was quantified using analysis of variance methods. RESULTS: Lung density declined linearly in the first few years of life and thereafter approximated adult values. Beginning with the anterior basal location, density at end inspiration (HU)=-835-(16 x age in years)+16 (if apical)+33 (if posterior)+23 (if medial)+14 (if lateral); standard error=38. At resting end exhalation =-616-(41 x age)+50 (if apical)+155 (if posterior)+74 (if medial)+46 (if lateral); standard error=68. CONCLUSION: We provide an initial basis for employing lung densitometry in infants and young children at end inspiration and resting end exhalation lung volumes. PMID- 15821936 TI - Intraurethral knot in a very-low-birth-weight infant: radiological recognition, surgical management and prevention. AB - We report a case where a knot developed in a urinary catheter and became lodged within the urethra of a very-low-birth-weight (VLBW) preterm infant. The catheter was removed with the assistance of a urologist. We recommend using caution when placing urinary catheters in VLBW infants and question the appropriateness of feeding tubes as catheters. Recognition on radiographs of malpositioned bladder catheters is vital to the care of these patients. All staff involved in the insertion, maintenance or removal of these catheters should be suitably trained to minimize the risk of knots and related complications. PMID- 15821937 TI - Multiple cis-regulatory elements and the yeast sulphur regulatory network are required for the regulation of the yeast glutathione transporter, Hgt1p. AB - HGT1 encodes a high-affinity glutathione transporter in the yeast Saccharomyces cerevisiae that is induced under sulphur limitation. The present work demonstrates that repression by organic sulphur sources is under the control of the classic sulphur regulatory network, as seen by the absence of expression in a met4delta background. Cysteine appeared to be the principal regulatory molecule, since elevated levels were seen in str4delta strains (deficient in cysteine biosynthesis) that could be repressed by elevated levels of cysteine, but not by methionine or glutathione. Investigations into cis-regulatory elements revealed that the previously described motif, a 9-bp cis element, CCGCCACAC, located at the -356 to -364 region of the promoter could in fact be refined to a 7-bp CGCCACA motif that is also repeated at -333 to -340. The second copy of this motif was essential for activity, since mutations in the core region of the second copy completely abolished activity and regulation by sulphur sources. Activity, but not regulation, could be restored by reintroducing an additional copy upstream of the first copy. A third region, GCCGTCTGCAAGGCA, conserved in the HGT1 promoters of the different Saccharomyces spp, was observed at -300 to 285 but, while mutations in this region did not lead to any loss in repression, the basal and induced levels were significantly increased. In contrast to a previous report, no evidence was found for regulation by the VDE endonuclease. The strong repression at the transport level by glutathione seen in strains overexpressing HGT1 was due to a glutathione-dependent toxicity in these cells. PMID- 15821938 TI - Chriz, a chromodomain protein specific for the interbands of Drosophila melanogaster polytene chromosomes. AB - Polytene interphase chromosomes are compacted into a series of bands and interbands reflecting their organization into independent chromosomal domains. In order to understand chromosomal organization, we set out to study the role of proteins that are selective for interbands. Here we describe the Drosophila melanogaster chromodomain protein Chriz that is coimmunoprecipitated with the zinc finger protein Z4. Both proteins colocalize exclusively to the interbands on Drosophila polytene chromosomes. Like Z4, Chriz is ubiquitously expressed throughout development and is associated with chromatin in all interphase nuclei. Following dissociation from chromatin, early in mitosis Chriz binds to the centrosomes and to the mitotic spindle. Newly induced amorphic Chriz alleles are early lethal, and ubiquitous overexpression of Chriz is lethal as well. Available Chriz hypomorphs which survive until pupal stage have a normal chromosomal phenotype. Reducing Z4 protein does not affect Chriz binding to polytene chromosomes and vice versa. Z4 is still chromosomally bound when Chriz protein is depleted by RNA interference. PMID- 15821939 TI - [Do physicians overestimate effects of acupuncture treatment?]. AB - BACKGROUND: Physicians' ratings about their patients' pain prove to be invalid compared to patients' ratings. This is especially true if pain rating acts as an indicator for therapy outcome. The aim of this study was to compare physicians' and patients' ratings of pain relief following acupuncture and to identify correlations between patient characteristics and potential miscalibrations. PATIENTS AND METHODS: In a cross-sectional study 291 pain patients with gonarthrosis or chronic low back pain and their attending physicians were asked to give their rating of patients' pain relief following acupuncture. Patients were interviewed by telephone, and doctors responded to questionnaires. RESULTS: The proportion of false-positive physicians' ratings was 81% referring to patients without self-reported benefit from acupuncture. Just every fifth patient without pain relief was correctly classified by his physician. There was no correlation between patients' characteristics and false-positive ratings of physicians. CONCLUSIONS: Evaluation of treatment in daily medical routine should be primarily based on information provided by patients. PMID- 15821940 TI - Solution 1H NMR study of the accommodation of the side chain of n-butyl-etiohemin I incorporated into the active site of cyano-metmyoglobin. AB - In order to identify the most readily deformable portion of the heme pocket in myoglobin, equine myoglobin was reconstituted with a meso-n-butyl substituent on centrosymmetric etiohemin-I. Solution 1H NMR data for the low-spin iron(III) cyanide complex of oxidized myoglobin that include 2D nuclear Overhauser enhancement spectroscopy contacts, paramagnetic relaxation, and dipolar shifts resulting from magnetic anisotropy show that the heme binds uniquely to the iron in a manner that arranges the methyl and ethyl substituents on a given pyrrole in a clockwise manner when viewed from the proximal side, and with the n-butyl group seated at the canonical alpha-meso position of native protohemin-IX. The butyl group is oriented sharply toward the proximal side and its protein contacts demonstrate that it is oriented largely into the "xenon hole" in myoglobin. The location of the n-butyl group on the proximal side near the vacancies places it within the region found to be most flexible in molecular dynamics simulation. A small, counterclockwise rotation of the pyrrole N-Fe-N vector of n-butyl etiohemin-I relative to that for native protohemin, indicated by both the prosthetic group methyl contact shift pattern and the prosthetic group contacts to heme pocket residues, is proposed to allow the xenon hole to accommodate better the n-butyl group. In contrast to previous work, which showed that a bulky polar substituent on etiohemin-I preferentially seats at the canonical gamma-meso position, the nonpolar n-butyl group selects the alpha-meso position. PMID- 15821941 TI - [Mitochondrial damage in human sperm caused by the antineoplastic agent betulinic acid]. AB - BACKGROUND AND OBJECTIVE: Human spermatozoa post-ejaculation show all elements of intrinsic (via mitochondria) and extrinsic (via death receptors) programmed cell death or apoptosis. One experimental therapeutic agent for malignant melanoma is betulinic acid (BA), a cytotoxic agent which induces intrinsic apoptosis via direct effects on mitochondria. To assess the potential side effects of systemic BA, its effects on motility, mitochondrial transmembrane potential (mTMP) and the apoptotic enzymes caspase-9 and -3, were monitored in human ejaculated spermatozoa. PATIENTS AND METHODS: Semen samples from 33 healthy volunteers were examined after incubation with 60 microg/mL betulinic acid for 5 and 60 minutes. RESULTS: Treatment with betulinic acid caused an immediate disruption of mitochondrial transmembrane potential and activation of the "death enzymes" caspase-9 and -3. The loss of mitochondrial potential was accompanied by a significant decrease of spermatozoal motility. CONCLUSION: Our results suggest that inducers of the mitochondrial pathway of apoptosis used in the treatment of malignant melanoma damage the sensitive mitochondria of spermatozoa. PMID- 15821942 TI - Epstein-Barr virus myocarditis as a cause of sudden death: two autopsy cases. AB - Although the Epstein-Barr virus (EBV) causes acute infection accompanied by a high fever in young people, there appears to be few reports of a fatal outcome involving myocarditis. We report two cases of unexpected sudden death due to acute myocarditis possibly caused by the EBV. They each visited a hospital due to common cold-like symptoms and unexpectedly died several days later. In both cases, autopsy revealed myocardial necrosis with marked lymphocytic infiltration. Polymerase chain reaction (PCR) screening was positive for the EBV, whereas immunohistochemistry and in situ hybridization for the EBV were negative. Serological investigations showed a mild elevation in antiviral capsid antigen IgG and anti-EBV nuclear antigen IgG in both cases. Immunohistochemical study of lymphocytic infiltrates showed strong positivity for a T-cell marker (CD45R0) in the myocardium and pharyngeal mucosa. These cases suggest the potential risk of mortality from acute EBV infection in young people, even without severe clinical manifestations, and the importance of microbiological investigations, including PCR procedures, in postmortem diagnosis of infectious diseases. PMID- 15821943 TI - Microbial forensics: the next forensic challenge. AB - Pathogens and toxins can be converted to bioweapons and used to commit bioterrorism and biocrime. Because of the potential and relative ease of an attack using a bioweapon, forensic science needs to be prepared to assist in the investigation to bring perpetrators to justice and to deter future attacks. A new subfield of forensics--microbial forensics--has been created, which is focused on characterization of evidence from a bioterrorism act, biocrime, hoax, or an inadvertent release. Forensic microbiological investigations are essentially the same as any other forensic investigation regarding processing. They involve crime scene(s) investigation, chain of custody practices, evidence collection, handling and preservation, evidence shipping, analysis of evidence, interpretation of results, and court presentation. In addition to collecting and analyzing traditional forensic evidence, the forensic investigation will attempt to determine the etiology and identity of the causal agent, often in a similar fashion as in an epidemiologic investigation. However, for attribution, higher resolution characterization is needed. The tools for attribution include genetic- and nongenetic-based assays and informatics to attempt to determine the unique source of a sample or at least eliminate some sources. In addition, chemical and physical assays may help determine the process used to prepare, store, or disseminate the bioweapon. An effective microbial forensics program will require development and/or validation of all aspects of the forensic investigative process, from sample collection to interpretation of results. Quality assurance (QA) and QC practices, comparable to those used by the forensic DNA science community, are being implemented. Lastly, partnerships with other laboratories will be requisite, because many of the necessary capabilities for analysis will not reside in the traditional forensic laboratory. PMID- 15821944 TI - Sclerosing encapsulating peritonitis: MRI diagnosis. PMID- 15821945 TI - [Value of color-coded duplex sonography in preoperative visualization of lower leg arteries for microsurgical fibula transfer]. AB - BACKGROUND: In patients being considered for a microsurgical fibular transfer, thorough examination of the vascular supply to the lower leg is indisputably necessary. The aim of this study was to evaluate the validity of color-coded duplex sonography (CCDS) in the assessment of the arteries of the lower leg. PATIENTS AND METHODS: In 13 patients needing bony reconstruction of the jaw 22 legs were examined using CCDS. The three arteries of the lower leg were visualized and traced from the ankle to the popliteal fossa. At every arterial segment Doppler sonographic examination was also performed. All 13 patients underwent digital subtraction arteriography (DSA) of the lower extremities, additionally. RESULTS: We were able to visualize and to assess all but one of the arteries using CCDS. In 13 extremities of 9 patients CCDS exhibited a normal vascular supply to the lower leg provided by three regular arteries. DSA confirmed this and the patients underwent fibular transfer. In the other 4 patients a regular vascular situation could not be shown by CCDS because of nonvisualization of arterial segments or pathological findings. Two of these patients received alternative bony transplants. In the remaining two patients angiography exhibited one leg to have a normal three vessel supply, respectively. Fibular transplants could be raised from this leg. In all patients CCDS was able to localize between 3 and 7 perforators branching off the peroneal artery. Their position determined the placement of the skin paddle to be raised with the bone. Further advantages of CCDS were the universal applicability to all patients and the possibility of performing it at our own department. The time required for the examination and the necessity of having adequate technical equipment were the disadvantages. CONCLUSION: In patients being considered for a microsurgical fibular transplantation, we recommend CCDS of the lower leg as a screening tool for the vascular supply of the lower leg. DSA and MRA are predominantly advocated for those patients, in whom CCDS reveals pathological or uncertain findings. PMID- 15821946 TI - Acute transient cerebellar dysfunction and stuttering following mild closed head injury. AB - INTRODUCTION: A wide range of pathologies can cause cerebellar dysfunction but there have been few reports of transient dysfunction after mild head injury. There are none describing stuttering as an acute symptom after such injuries. CASE REPORT: We report a 12-year-old boy who presented with headache, stuttering speech and truncal and bilateral peripheral cerebellar dysfunction after falling prone from his skateboard. Computed tomographic brain imaging showed normal results. All clinical symptoms and signs had resolved within 24 h obviating further investigation. A follow up clinical interview and examination 4 weeks later revealed no recurrence of cerebellar or speech dysfunction. CONCLUSION: This is the first reported case of acute transient cerebellar dysfunction and stuttering after mild closed head injury. Neurogenic causes for stuttering are increasingly being recognised. PMID- 15821947 TI - Familial Chiari type I malformation with syringomyelia in two siblings: case report and review of the literature. AB - OBJECTIVE: Familial cases of Chiari malformation with syringomyelia are rare. The majority of the reported series and case reports detail sporadic cases. The authors report two siblings who presented with Chiari type I malformation and syringomyelia (CMI+S). CLINICAL PRESENTATION: We report two sisters who each presented with scoliosis on routine school physicals. Their clinical examination was unremarkable; however, imaging studies demonstrated a Chiari malformation with syringomyelia. Both underwent cervicomedullary decompression, and follow-up imaging studies revealed resolution of the syringomyelia. CONCLUSION: A review of the literature reveals fewer than ten previous reports of familial CMI+S in the past 30 years. Although rare, the existence of familial cases of CMI+S suggests a genetic component to the pathogenesis of this condition in at least a proportion of patients. Neurosurgeons should be aware of the familial aggregation of CMI+S. PMID- 15821948 TI - Should orthopaedics be globalised? PMID- 15821949 TI - Locomotor activity detects subunit-selective effects of agonists and decahydroisoquinoline antagonists at AMPA/kainic acid ionotropic glutamate receptors in adult rats. AB - RATIONALE: In vitro studies have identified a series of decahydroisoquinoline compounds with differential selectivity for the subunits that comprise AMPA/kainic acid receptors. Compounds have been identified that have preferential activity at AMPA receptors (LY302679), whereas others (LY377770) have affinity for GluR5-kainic acid preferring subunit, which is activated by ATPA and kainic acid. OBJECTIVES: These studies set out to determine if locomotor activity could differentiate these profiles in vivo. METHODS: Locomotor activity was assessed in photocell drums in male Lister Hooded rats. RESULTS: AMPA, kainic acid and the GluR5 selective agonist ATPA, all suppressed spontaneous locomotor activity (SLA) in rats at doses of 1.0, 5.0 and 20 mg/kg resp. All three agonists achieve micromolar concentrations measured in whole brain after dosing with 10 mg/kg SC. The decahydroisoquinoline antagonist compounds, LY302679 (GluR2), LY293558 (GluR2, 5) and LY377770 (GluR5) all decreased SLA in rats (ED(min) 2.5, 5.0 and 20 mg/kg respectively). The rank order of potency at GluR2 subunits (LY302679>LY293558>LY377770) was reflected in the same rank order of activity for suppression of SLA. LY293558 reversed the suppression of SLA induced by all three agonists (0.62--2.5 mg/kg). LY377770 reversed the effects of ATPA only (ED(min) 1.0 mg/kg), LY302679 (ED(min) 2.5 mg/kg) attenuated the effect of kainic acid but was ineffective against AMPA and ATPA. CONCLUSIONS: Both agonist and antagonist suppression of SLA is associated with greater affinity for the GluR2 subunit, while compounds with affinity for the GluR5 subunit were less potent in suppressing SLA. PMID- 15821950 TI - Effects of mGlu1 receptor blockade on anxiety-related behaviour in the rat lick suppression test. AB - RATIONALE: Group I metabotropic glutamate receptor antagonists, which block both the mGlu1 and mGlu5 receptors, have been shown to have anxiolytic effects in the lick suppression test in rats. OBJECTIVE: The anxiolytic potential of the selective mGlu1 antagonist 3,4-dihydro-2H-pyrano[2,3]beta-quinolin-7-yl)(cis-4 methoxycyclohexyl)methanone (JNJ16259685) was investigated and compared with the mGlu5 antagonist MPEP. METHODS: Anxiety-related behaviour was assessed in lick suppression and in the elevated zero maze in rats. Non-specific effects on pain threshold, water intake and locomotor activity were also measured. RESULTS: Acute administration of JNJ16259685 or MPEP increased the number of licks (lowest active dose 2.5 mg/kg IP for each compound). JNJ16259685 did not increase water intake or reduce acute pain threshold, suggesting that the anxiolytic-like properties are specific. However, acute administration decreased locomotor activity. The effects of chronic administration of JNJ16259685 over 14 days (5 mg/kg bid) on lick suppression were comparable to those seen after acute administration, arguing against development of behavioural tolerance or sensitisation. Yet, there was a tendency for an increase in locomotor activity after cessation of chronic treatment. Acute co-administration of both JNJ16259685 and MPEP had additive effects on the number of licks. No anxiolytic-like properties of JNJ16259685 were observed in the elevated zero maze. CONCLUSION: Our data suggest that the anxiolytic-like effects induced by group I metabotropic glutamate receptor antagonists are mediated through both mGlu1 and mGlu5 receptors. Rather than producing a general anxiolytic-like effect, the effects seen following mGlu1 antagonism seem task-dependent, as prominent effects were seen in a conflict procedure, but not in a task based on spontaneous exploration. PMID- 15821951 TI - Effects of a metabotropic glutamate(2/3) receptor agonist (LY544344/LY354740) on panic anxiety induced by cholecystokinin tetrapeptide in healthy humans: preliminary results. AB - RATIONALE: Preclinical findings have repeatedly shown an anxiolytic-like action of agonists at metabotropic glutamate receptors type II, such as LY354740. OBJECTIVE: We aimed to investigate the effect of LY544344, the prodrug of LY354740, upon experimental panic anxiety in humans. METHODS: Twelve healthy human volunteers were treated orally with 80 mg bid LY544344 for 1 week in a randomized placebo-controlled cross-over study before 50 mug cholecystokinin tetrapeptide (CCK-4) was injected intravenously. We assessed CCK-induced panic and anxiety symptoms and measured stress hormone release. RESULTS: While no significant treatment effect emerged in the entire sample, a significant reduction of the number of CCK-4-induced panic symptoms and of CCK-4-induced subjective anxiety ratings was detected after removing two subjects who did not show decreased CCK-4-elicited adrenocorticotropin (ACTH) release after LY544344 compared to placebo treatment. CONCLUSIONS: Further studies are needed to clarify the potential of LY544344 as a new anxiolytic or antipanic drug. PMID- 15821952 TI - Effect of the blockade of mu1-opioid and 5HT2A-serotonergic/alpha1-noradrenergic receptors on sweet-substance-induced analgesia. AB - RATIONALE: Sweet-substance-induced analgesia has been widely studied, and the investigation of the neurotransmitters involved in this antinociceptive process is an important way for understanding the involvement of the neural system controlling this kind of antinociception. OBJECTIVE: The aim of this study was to investigate the involvement of opioid and monoaminergic systems in sweet substance-induced analgesia. METHODS: The present work was carried out in an animal model with the aim of investigating whether acute (24 h) or chronic (14 days) intake of a sweet substance, such as sucrose (250 g/l), is followed by antinociception. Tail withdrawal latencies in the tail-flick test were measured before and immediately after this treatment. Immediately after the recording of baseline values, independent groups of rats were submitted to sucrose or tap water intake and, after chronic treatment, they were pretreated with intraperitoneal administration of (1) naltrexone at 0.5, 1, 2 or 3 mg/kg; (2) naloxonazine at 5, 10, 20 or 30 mg/kg; (3) methysergide at 0.5, 1, 2 or 3 mg/kg; (4) ketanserin at 0.5, 1, 2 or 3 mg/kg; or (5) physiological saline. RESULTS: Naltrexone and methysergide at two major doses decreased sweet-substance-induced analgesia after chronic intake of a sweet substance. These effects were corroborated by peripheral administration of naloxonazine and ketanserin. CONCLUSIONS: These data give further evidence for: (a) the involvement of endogenous opioids and a mu1-opioid receptor in the sweet-substance-induced antinociception; (b) the involvement of monoamines and 5HT2A serotonergic/alpha1 noradrenergic receptors in the central regulation of the sweet-substance-produced analgesia. PMID- 15821953 TI - Lorazepam reinstates punishment-suppressed remifentanil self-administration in rats. AB - RATIONALE: We recently described a reinstatement procedure that models relapse to drug abuse in cases where abstinence results from aversive consequences of drug use. The potential value of this punishment-based model of relapse depends on its sensitivity to relapse-inducing events that are ineffective in the widely used extinction-based model. OBJECTIVES: It is known that certain drugs can have anti punishment effects, but these drugs have not been tested in the punishment-based reinstatement procedure. Therefore, the effects of the benzodiazepine, lorazepam, were examined using punishment-based and extinction-based reinstatement procedures. METHODS: Rats self-administered the opioid, remifentanil (4 microg/kg per infusion). Two punishment groups were trained with response-contingent footshock that suppressed baseline rates of responding to zero. In an extinction group, remifentanil delivery was discontinued, and baseline responding stabilized at a low rate (mean=0.06 responses/min). Lorazepam (0.08-10 mg/kg, IP) was given during test sessions with the shock contingency discontinued for both punishment groups. Remifentanil delivery was maintained during testing in one punishment group but not the other. RESULTS: Lorazepam reinstated self-administration responding in both punishment groups but not in the extinction group. Priming injections of heroin reinstated responding in both the punishment and extinction groups, but combining heroin and lorazepam did not enhance reinstatement. CONCLUSIONS: This is the first demonstration that a trigger for relapse may have different effects depending on whether aversive conditioning contributed to the achievement of abstinence. It may be important to consider potential anti punishment effects of both abused drugs and therapeutic agents in the treatment of individuals with a history of drug abuse. PMID- 15821954 TI - Role of the pedunculopontine tegmental nucleus in sensorimotor gating and reward related behavior in rats. AB - RATIONALE: The pedunculopontine tegmental nucleus (PPTg) is involved in the execution and regulation of a variety of behaviors. Most investigations used brain lesions that have certain disadvantages, such as functional compensation over time. OBJECTIVES: In the present study, we investigated by temporary, reversible inhibition of neurons the role of the PPTg in sensorimotor gating, measured as prepulse inhibition (PPI) of the acoustic startle response (ASR) using variable interstimulus intervals (ISI). In a second set of experiments we examined by the same technique the role of the PPTg in a progressive-ratio instrumental response task. METHODS: Local infusions of the GABA(A)-receptor agonist muscimol (0.05 microg and 0.5 microg/0.3 microl, or vehicle) were applied through indwelling microinfusion cannulae into the PPTg of freely moving rats. ASR and PPI were measured using acoustic stimuli of 100 dB (pulse) and 80 dB (prepulse) using ISIs of 25, 120, 520 and 1,020 ms. Instrumental behavior (lever pressing for casein pellets) was assessed in a Skinner box. Motor activity was measured in an open field. RESULTS: Intra-PPTg infusions of muscimol dose dependently attenuated PPI at ISIs of 120 ms and 520 ms, but not at longer or shorter ISIs. ASR magnitude in pulse-alone trials was not significantly affected. Intra-PPTg infusion of 0.5 microg muscimol reduced the break point of instrumental responding (testing sequence where the rats fail to respond according to an increased ratio of reinforcement). No effects on food-preference and open-field activity were found. CONCLUSIONS: These findings suggest that GABAergic neurotransmission in the PPTg plays an important role for sensorimotor gating at intermediate ISIs and for response selection under demanding schedules of reinforcement. PMID- 15821955 TI - Surgical and pharmacological suppression of glucocorticoids prevents the enhancement of morphine conditioned place preference by uncontrollable stress in rats. AB - RATIONALE: Stress and one of the physiological components of most stress responses, glucocorticoid hormones (CORT), are known to influence the rewarding effects of a number of drugs of abuse. We have previously shown that an acute uncontrollable stressor (inescapable shock, IS) potentiates the rewarding effects of morphine using conditioned place preference (CPP). OBJECTIVES: The following experiments were conducted to determine the role of CORT in this process. METHODS: First, the CORT response to 3.0 mg/kg morphine was measured in male Sprague-Dawley rats 24 h following exposure to IS. Second, we determined the effect of adrenalectomy (ADX) on the IS-potentiated CPP to morphine. Finally, we used the temporary CORT synthesis inhibitors metyrapone and aminoglutethimide to determine the necessity of CORT rises during either IS or morphine administration on the potentiated CPP response. RESULTS: Prior IS significantly potentiated the CORT response to morphine. ADX significantly blocked the potentiated CPP to morphine produced by previous IS. However, CORT inhibition during IS had no effect on the IS potentiation of morphine CPP, whereas inhibition during morphine administration completely blocked this potentiation. CONCLUSIONS: The results indicate that the CORT response to morphine is enhanced in rats that were previously exposed to an uncontrollable stressor, and that this response to the drug, not the stressor, is necessary for the stress-enhanced potentiation of morphine CPP. PMID- 15821956 TI - Implication of 5-HT2 receptor subtypes in the mechanism of action of antidepressants in the four plates test. AB - RATIONALE: The selective serotonin reuptake inhibitors (SSRIs) and the serotonin and noradrenaline reuptake inhibitors (SNRIs) increase synaptic levels of serotonin, leading to an increased activation of a multitude of specific postsynaptic 5-HT receptors. However, it is not yet known which 5-HT receptor subtypes mediate the therapeutic effects of antidepressants. METHODS: The effects of the SSRI, paroxetine and the SNRI, venlafaxine were evaluated in the mouse four plates test (FPT). RESULTS: Paroxetine administered intraperitoneally (IP) (0.5, 2-8 mg/kg) potently augmented the number of punished passages accepted by mice in this paradigm. The effects of paroxetine (8 mg/kg) were not reversed by the selective 5-HT2C receptor antagonist, RS 10-2221 (0.1 mg/kg and 1 mg/kg) or the selective 5-HT2B/2C receptor antagonist SB 206553 (0.1 mg/kg and 1 mg/kg), at doses which lack an effect when administered alone. In contrast, the selective 5 HT2A receptor antagonist, SR 46349B (0.1 mg/kg and 1 mg/kg) completely abolished the paroxetine-induced increase in punished passages. The acute administration of venlafaxine induced an anxiolytic-like effect in the FPT at the doses of 2-16 mg/kg. This effect was reversed by the 5-HT2B/2C receptor antagonist as did SR 46349B, for both doses administered. Our results strongly suggest that activation of 5-HT2A receptors is critically involved in the anxiolytic activity of paroxetine, whereas the 5-HT2A and 5-HT2B receptors are involved in the anti punishment action of venlafaxine in the FPT. The co-administration of selective 5 HT2A, 2B, 2C receptor agonists (DOI, 0.06 mg/kg and 0.25 mg/kg; BW 723C86, 0.5 mg/kg and 2 mg/kg and RO 60-0175, 0.06 mg/kg and 0.25 mg/kg), respectively, was subsequently investigated. The effects of sub-active doses of paroxetine (0.25 mg/kg and 1 mg/kg) were augmented by BW 723C86 and RO 60-0175 receptor agonist challenge. The anti-punishment effects of venlafaxine (0.25 mg/kg and 1 mg/kg) were potentialised only by DOI co-administration. CONCLUSION: These results indicate that the co-administration of 5-HT2 receptor agonists with paroxetine and venlafaxine may provide a powerful tool for enhancing the clinical efficacy of these antidepressants. PMID- 15821957 TI - The cannabinoid CB1 receptor antagonist SR141716A reduces appetitive and consummatory responses for food. AB - RATIONALE: The CB1 receptor antagonist SR141716A reduces food intake in rats. This effect is likely to depend on modulation of reward related processes. OBJECTIVE: To investigate the effects of SR141716A on responding for food under a second order instrumental task in which responding and consumption of food can be separated, and on Pavlovian responding for a stimulus predictive of food reward. METHODS: Instrumental responding and pellet consumption following administration of SR141716A (0-3 mg/kg) were recorded under an FI5 min FR5(5:S) operant schedule that incorporates both a 5 min initial appetitive phase and a 25 min consummatory phase. We compared the drug-induced change in responding to that recorded following a reduction in motivational state induced by pre-feeding. In a second experiment we assessed the effects of SR141716A (0-3 mg/kg) on Pavlovian approach behaviour for a stimulus (lever) associated with food reward (CS+) and a neutral stimulus (lever) not associated with reward (CS-). RESULTS: SR141716A reduced pellet consumption and instrumental responding during both the appetitive and consummatory phases of the second order schedule. Pre-feeding had a similar effect on responding during the appetitive phase, suggesting an effect on incentive motivation. SR141716A also blocked an enhancement of responding that occurred during the consummatory phase in pre-fed animals. SR141716A and pre feeding had no effect on responding in the Pavlovian autoshaping paradigm. CONCLUSIONS: SR141716A impacts on motivational processes in both the appetitive and consummatory phases of feeding behaviour. PMID- 15821958 TI - Stable expression of constitutively activated mutant h5HT6 and h5HT7 serotonin receptors: inverse agonist activity of antipsychotic drugs. AB - RATIONALE: In order to determine the possible relationship between antipsychotic drug properties and inverse agonist activity at h5HT6 and h5HT7 receptors, constitutively activated forms of these receptors were created by site-specific mutagenesis. Typical and atypical antipsychotic drugs were assayed for their potencies as inverse agonists at these mutated receptors. OBJECTIVES: Stable cell lines expressing constitutively activated forms of the h5HT6 and h5HT7 receptors were created. Typical and atypical antipsychotic drugs demonstrating high to moderate affinities for the h5HT6 and h5HT7 receptors were assayed for their potencies in reversing the agonist-independent activity (inverse agonist activity). RESULTS: The E322R h5HT6 mutant and the S267K h5HT7 mutant displayed sufficiently robust agonist-independent activity when expressed in stable cell lines to allow the detailed, concentration-dependent, investigation of the inverse agonist activity of typical and atypical antipsychotic drugs. All the drugs tested displayed inverse agonist activity at both the activated h5HT6 and h5HT7 receptors. Native forms of these receptors did not display any constitutive activity. Interestingly, atypical antipsychotic drugs displayed potent inverse agonist activity, relative to typical antipsychotic drugs, at the h5HT7 receptor. LSD displayed neutral antagonist properties at the mutant h5HT7 receptor. CONCLUSIONS: Site-specific mutations in the third intracellular loop of the G(s) coupled h5HT6 and h5HT7 receptors produce constitutive activation. Antipsychotic drugs display inverse agonist activity at the activated receptors. The inverse agonist mechanism-of-action of the atypical antipsychotic drugs at the h5HT7 receptors may be different from the typical antipsychotic drugs as these drugs displayed far higher potencies as inverse agonists at the h5HT7 receptor. PMID- 15821959 TI - CB1 cannabinoid receptor agonists increase intracranial self-stimulation thresholds in the rat. AB - RATIONALE: Addictive drugs have a number of commonalities in animal behavioral models. They lower intracranial self-stimulation (ICSS) thresholds, support self administration, and produce conditioned place preference (CPP). However, cannabinoids appear atypical as drugs of abuse, since there are controversial data in the literature concerning their reinforcing properties. OBJECTIVES: The aim of the present study was to examine the effects of cannabinoids on brain reward using the rate-frequency curve shift paradigm of ICSS. METHODS: Male Sprague-Dawley rats were implanted with electrodes into the medial forebrain bundle (MFB). Rate-frequency functions were determined by logarithmically decreasing the number of cathodal pulses in a stimulation train from a value that sustained maximal responding to one that did not sustain responding. After brain stimulation reward thresholds stabilized rats received intraperitoneal (IP) injections of the potent CB1 receptor agonists WIN 55,212-2 (graded doses 0.1, 0.3, 1 and 3 mg/kg), CP 55,940 (graded doses 10, 30, 56 and 100 microg/kg), or HU 210 (graded doses 10, 30, 100 microg/kg). RESULTS: With the exception of the highest dose of all cannabinoid agonists tested, which significantly increased the threshold frequency required for MFB ICSS, all other doses of the tested drugs did not affect ICSS thresholds. The CB1 receptor antagonist SR141716A reversed the actions of WIN 55,212-2 and CP 55,940, but not HU-210. However, the selective CB1 cannabinoid receptor antagonist AM 251 counteracted the effect of HU-210. Both CB1 receptor antagonists, at the doses used in the present study, did not affect reward thresholds by themselves. CONCLUSIONS: The present results indicate that cannabinoid agonists do not exhibit reinforcing properties in the ICSS paradigm, but rather have an inhibitory influence on reward mechanisms. The results suggest that the anhedonic effects of cannabinoids are probably mediated by cannabinoid CB1 receptors. PMID- 15821961 TI - Diagnosis of Alzheimer's disease using brain perfusion SPECT and MR imaging: which modality achieves better diagnostic accuracy? AB - PURPOSE: The purpose of this study was to compare the accuracy of MR imaging and brain perfusion single-photon emission tomography (SPECT) in diagnosing Alzheimer's disease (AD). METHODS: The transaxial section display of brain perfusion SPECT, three-dimensional stereotactic surface projection (3D-SSP) SPECT image sets, thin-section MR imaging of the hippocampus and perfusion MR imaging were evaluated in 66 subjects comprising 35 AD patients and 31 subjects without AD. SPECT and MR imaging were visually interpreted by two experts and two novices, and the diagnostic ability of each modality was evaluated by receiver operating characteristic (ROC) analysis. RESULTS: In the experts' interpretations, there was no significant difference in the area under the ROC curve (Az) between 3D-SSP and thin-section MR imaging, whereas the Az of transaxial SPECT display was significantly lower than that of 3D-SSP (3D-SSP: 0.97, thin-section MR imaging: 0.96, transaxial SPECT: 0.91), and the Az of perfusion MR imaging was lowest (0.63). The sensitivity and specificity of each modality were, respectively, 80.0% and 96.8% for 3D-SSP, 77.1% and 96.8% for thin section MR imaging, 60.0% and 93.5% for transaxial SPECT display and 34.3% and 100% for perfusion MR imaging. In the novices' interpretations, the Az, sensitivity and specificity of 3D-SSP were superior to those of thin-section MR imaging. CONCLUSION: Thin-section hippocampal MR imaging and 3D-SSP image sets had potentially equivalent value for the diagnosis of AD, and they were superior to transaxial SPECT display and perfusion MR imaging. For avoidance of the effect of interpreters' experience on image evaluation, 3D-SSP appears to be optimal. PMID- 15821962 TI - Survival benefit after revascularization is independent of left ventricular ejection fraction improvement in patients with previous myocardial infarction and viable myocardium. AB - PURPOSE: This study was designed to assess the relationships among myocardial viability, changes in left ventricular (LV) ejection fraction after coronary revascularization and long-term event-free survival in patients with previous myocardial infarction and LV dysfunction. METHODS: We studied 253 patients with previous myocardial infarction and evidence of dysfunctional viable myocardium as assessed by echocardiography and 99mTc-sestamibi imaging. Coronary revascularization was performed in 142 patients, while 111 were medically treated. In revascularized patients, echocardiography was repeated 12 months later to detect LV ejection fraction improvement, defined as an increase of > or =5% compared with baseline. All patients were followed for a mean period of 52+/ 29 months. Cardiac death and non-fatal myocardial infarction were considered as events. RESULTS: Event-free survival was higher in revascularized than in medically treated patients (P<0.001). Ejection fraction increased by > or =5% in 82 (58%) revascularized patients, and the extent of viable myocardium was the strongest predictor of such improvement (P<0.001). Event-free survival was similar for patients with (n=82) and patients without (n=60) LV ejection fraction improvement after revascularization, and it was better in revascularized than in medically treated patients in the presence of either substantial (> or =5 viable segments) or low-intermediate (1-4 viable segments) viability (both P<0.01). CONCLUSION: In patients with previous myocardial infarction and evidence of viable myocardium, coronary revascularization procedures improve outcome at long term follow-up independently of LV ejection fraction improvement. PMID- 15821963 TI - Influence of the polyol pathway on norepinephrine transporter reduction in diabetic cardiac sympathetic nerves: implications for heterogeneous accumulation of MIBG. AB - PURPOSE: Cardiac scintigraphic studies using (123)I-labeled metaiodobenzylguanidine ([(123)I]MIBG) have demonstrated heterogeneous myocardial accumulation of MIBG in diabetes. The accumulation has been found to correlate with a heterogeneous decrease in the expression of norepinephrine transporter (NET). In diabetic peripheral nerve tissue, polyol pathways are activated and cause nerve dysfunction and degeneration. However, there has been little research on the polyol pathway and cardiac sympathetic nerves. Therefore, to assess the influence of the polyol pathway on cardiac sympathetic nervous function, we investigated the regional accumulation of MIBG and NET protein expression in diabetic model rats treated with aldose reductase inhibitor (ARI) for the blockade of polyol pathways. METHODS: Rats were given a single intravenous injection of streptozotocin (n=76, STZ-D rats). Starting the day after STZ injection, ARI was administered daily to 42 of the rats for 4 weeks (ARI-D rats). To assess the cardiac sympathetic nervous function, [(125)I]MIBG autoradiographic experiments were carried out. Finally, NET protein expression was assessed with a saturation binding assay. RESULTS: The myocardial sorbitol concentration was significantly higher in STZ-D rats than in ARI-D rats. There was no heterogeneous accumulation of MIBG in ARI-D rats. There was a heterogeneous decrease of NET expression in STZ-D rats, but not in ARI-D or control rats. CONCLUSION: The gathered data indicate that the enhanced polyol pathway correlates with the decrease in regional cardiac sympathetic nervous function, and this impairment may lead to the reduction of NET protein in cardiac sympathetic nerves of the diabetic inferior wall. PMID- 15821964 TI - Hyperpolarized 3He MRI and 81mKr SPECT in chronic obstructive pulmonary disease. AB - PURPOSE: During recent years, magnetic resonance imaging (MRI) using hyperpolarised (HP) 3He gas has emerged as a promising new method for the imaging of lung ventilation. However, systematic comparisons with nuclear medicine techniques have not yet been performed. The aim of this study was to compare ventilation imaging methods in 26 patients with chronic obstructive pulmonary disease (COPD) and nine lung healthy volunteers. METHODS: HP 3He MRI, 81mKr single-photon emission computed tomography (SPECT), high-resolution computed tomography (HRCT) and pulmonary function tests were performed. The three scans were scored visually as percentage of non-ventilated/diseased lung, and a computer-based objective measure of the ventilated volume in HP 3He MRI and 81mKr SPECT and an emphysema index in HRCT were calculated. RESULTS: We found a good correlation between HP 3He MRI and 81mKr SPECT for both visual defect score (r=0.80, p<0.0001) and objective estimate of ventilation (r=0.45, p=0.0157). In addition, both scans were well correlated with reference methods for the diagnosis of emphysema (pulmonary function test and HRCT). The defect scores were largest on 81mKr SPECT (the score on HP 3He MRI was one-third less than that on 81mKr SPECT), but the difference was reduced after normalisation for different breathing depths (HP 3He MRI at total lung capacity; 81mKr SPECT at tidal breathing at functional residual capacity). CONCLUSION: HP 3He MRI provides detailed ventilation distribution images and defect scores are comparable on HP 3He MRI and 81mKr SPECT. Additionally, new insights into the regional pulmonary microstructure via the apparent diffusion coefficient measurements are provided by HP 3He MRI. HP 3He MRI is a promising new diagnostic tool for the assessment of ventilation distribution. PMID- 15821965 TI - Validation of prospective whole-body bone marrow dosimetry by SPECT/CT multimodality imaging in (131)I-anti-CD20 rituximab radioimmunotherapy of non Hodgkin's lymphoma. AB - PURPOSE: Radioimmunotherapy (RIT) for relapsed non-Hodgkin's lymphoma is emerging as a promising treatment strategy. Myelosuppression is the dose-limiting toxicity and may be particularly problematic in patients heavily pretreated with chemotherapy. Reliable dosimetry is likely to minimise toxicity and improve treatment efficacy, and the aim of this study was to elucidate the complex problems of dosimetry of RIT by using an integrated SPECT/CT system. METHODS: As a part of a clinical trial of (131)I-anti-CD20 rituximab RIT of non-Hodgkin's lymphoma, we employed a patient-specific prospective dosimetry method utilising the whole-body effective half-life of antibody and the patient's ideal weight to calculate the administered activity for RIT corresponding to a prescribed radiation absorbed dose of 0.75 Gy to the whole body. A novel technique of quantitation of bone marrow uptake with hybrid SPECT/CT imaging was developed to validate this methodology by using post-RIT extended imaging and data collection. RESULTS: A strong, statistically significant correlation (p=0.001) between whole body effective half-life of antibody and effective marrow half-life was demonstrated. Furthermore, it was found that bone marrow activity concentration was proportional to administered activity per unit weight, height or body surface area (p<0.001). CONCLUSION: The results of this study show the proposed whole body dosimetry method to be valid and clinically applicable for safe, effective RIT. PMID- 15821966 TI - Visualisation of impaired dopamine biosynthesis in a case of aromatic L-amino acid decarboxylase deficiency by co-registered 18F-FDOPA PET and magnetic resonance imaging. PMID- 15821967 TI - Acquired ileal atresia in infancy: report of two cases. AB - Two infants, aged 3 months and 4 months, with acquired ileal atresia in the distal ileum are described. Both of them presented with features of intestinal obstruction following a diarrhoeal episode. In both patients the time interval between the onset of symptoms and detection of atresia at surgery was very short, i.e. 2 weeks. In our opinion, strangulation with subsequent resorption of gangrenous gut could have been the underlying mechanism in both patients. However, a short interval between initiation of symptomatology and detection of atresia prompted us to postulate a thromboembolic phenomenon as the cause of these lesions. While in one patient intussusception was noted on sonography, in the other there was no clear-cut aetiology present except for a high-lying caecum with a long mesentery that might have led to volvulus. We have reviewed the pertinent literature of this very rare sequela of gut inflammation. PMID- 15821968 TI - Compensatory brow asymmetry: anatomic study and clinical experience. AB - To obtain detailed information about the interdigitation between the frontalis muscle and the orbicularis oculi muscle, the authors performed an anatomic study involving 9 cadavers (17 specimens). Three types of interdigitation were recognized. Complete interdigitation was noted in 13 specimens, and partial interdigitation in 13 specimens. In one specimen, there was no interdigitation between the frontalis muscle and the orbicularis oculi muscle. Although most of the specimens showed complete interdigitation between the frontalis muscle and the orbicularis oculi muscle, the interdigitation area exhibited architectural heterogeneity. The study findings provide a basis for a better understanding of compensatory eyebrow ptosis. In addition, two cases supporting the theory that compensatory eyebrow asymmetry may resolve when the palpebral ptosis is treated are presented. PMID- 15821969 TI - Anterior scoring of the upper helical cartilage as a refinement in aesthetic otoplasty. AB - BACKGROUND: Anterior scoring with the use of simple or dedicated instruments, toothed forceps, endoscopic carpal tunnel release instruments, and needles has been described previously. The upper third of the ear easily maintains the original shape because memory and elasticity are stronger than in the middle or the lower part of the ear. This report describes a further refinement to the Chong-Chet anterior scoring technique, consisting of anterior scoring of the upper helical cartilage to correct the helical radix upper prominence. METHODS: A retrospective study analyzed 20 surgeries for prominent ears. All the subjects had undergone otoplasty softening the helix. RESULTS: There were no residual ear prominence/upper third prominence or cartilage irregularities at the 1-year follow-up evaluation. Anterior auricular cartilage scoring is an effective technique for controlling the degree and position of the antihelical fold. Furthermore, the upper third of the pinna seems to have a stronger memory than the middle third. To prevent this late complication the authors routinely perform anterior scoring up to the helix, weakening that cartilage usually untouched with other procedures. CONCLUSION: This refinement, in combination with other procedures is safe, easy, and fast, giving reproducible and good aesthetic results. PMID- 15821971 TI - Management of enophthalmos and superior sulcus deformity induced by the silent sinus syndrome. AB - BACKGROUND: Silent sinus syndrome is a dysfunction of the maxillary sinus that induces a progressive and asymptomatic enophthalmos with prominent deep superior sulcus deformity. METHODS: Two cases of silent sinus syndrome are reported, and the simultaneous management of both enophthalmos and superior sulcus deformity caused by this syndrome is discussed. RESULTS: The patients underwent surgical endoscopic maxillary meatotomy and transconjunctival subperiosteal implantation of porous polyethylene sheets. The treatment successfully corrected both the enophthalmos and the upper eyelid sulcus deformity. However, small degrees of vertical eye dystopia were observed. CONCLUSIONS: Silent sinus syndrome is a rare cause of enophthalmos and superior sulcus deformity. Orbital floor implants can be used to increase the volume of the orbital contents, but vertical eye dystopia is likely to be induced if this method of treatment is the only option chosen. PMID- 15821973 TI - The bovine immunodeficiency virus: cloning of a tat/rev cDNA encoding a novel Tat protein with enhanced transactivation activity. AB - Previous studies have shown that BIV may encode two types of Tat proteins of 103 and 108 amino acids, respectively. Here, we report the characterization of a new BIV Tat protein (Tat236) derived from a tat/rev cDNA. The tat/rev cDNA was obtained by reverse transcription-PCR from RNA extracted from cells infected with BIV. BIV was rescued by cell co-cultivation from the spleen of rabbits exposed for 3 years to the R29 isolate of BIV. Sequence analysis indicated that BIV Tat236 contains the first 98 amino acids of Tat103 and the 3' end 138 amino acids of Rev. Reporter gene assays indicated that transactivation of BIV long terminal repeat (LTR) by Tat236 is higher than by the original BIV Tat proteins in several cell types. By using overlapping deletion mutants, evidence was given that the predicted basic domain of Rev within Tat236 plays a major role in the observed enhanced transactivation activity of the protein. However, the intact functional domain of the original BIV Tat is required for efficient transactivation. This is the first report of a hybrid Tat protein from BIV or any lentiviruses that shows higher transactivation than the original transactivator Tat proteins. PMID- 15821974 TI - The question of Citrus viroid IV as a cocadviroid. AB - It has been suggested that Citrus viroid IV (CVd-IV) be classified as a species within the genus Cocadviroid. This relationship was based on the presence of a terminal conserved hairpin (TCH) and absence of a terminal conserved region (TCR) as specific structural motifs in common with isolates of Coconut cadang-cadang viroid (CCCVd) as well as phylogenetic relationships with members of the genus Cocadviroid. Evidence is presented for a "vestigial" TCR in CVd-IV as well as the introduction of the terminal repeat region (TRR) motif and an alternative sequence analysis that suggests a closer phylogenetic relationship of CVd-IV to isolates of Citrus exocortis viroid (CEVd), a species within the genus Pospiviroid than to CCCVd. This position is further supported by the striking similarity of biological properties between CVd-IV and CEVd with the suggestion offered that biological evidence be considered for specific adjustments to any overall classification scheme for viroids. PMID- 15821975 TI - Linkage disequilibrium and association studies in higher plants: present status and future prospects. AB - During the last two decades, DNA-based molecular markers have been extensively utilized for a variety of studies in both plant and animal systems. One of the major uses of these markers is the construction of genome-wide molecular maps and the genetic analysis of simple and complex traits. However, these studies are generally based on linkage analysis in mapping populations, thus placing serious limitations in using molecular markers for genetic analysis in a variety of plant systems. Therefore, alternative approaches have been suggested, and one of these approaches makes use of linkage disequilibrium (LD)-based association analysis. Although this approach of association analysis has already been used for studies on genetics of complex traits (including different diseases) in humans, its use in plants has just started. In the present review, we first define and distinguish between LD and association mapping, and then briefly describe various measures of LD and the two methods of its depiction. We then give a list of different factors that affect LD without discussing them, and also discuss the current issues of LD research in plants. Later, we also describe the various uses of LD in plant genomics research and summarize the present status of LD research in different plant genomes. In the end, we discuss briefly the future prospects of LD research in plants, and give a list of softwares that are useful in LD research, which is available as electronic supplementary material (ESM). PMID- 15821976 TI - Small heat shock proteins are differentially regulated during pollen development and following heat stress in tobacco. AB - In plants small heat shock proteins (sHsp) are abundantly expressed upon heat stress in vegetative tissue, however, sHsp expression is also developmentally induced in pollen. The developmental induction of sHsp has been related to the potential for stress-induced microspore embryogenesis. We investigated the polymorphism among sHsp and their expression during pollen development and after heat stress in tobacco. Real-time RT-PCR was used for quantification of mRNA of two known and nine newly isolated cDNAs representing cytosolic sHsp. At normal temperature most of these genes are not transcribed in vegetative tissues, however, all genes were expressed during pollen development. Low levels of mRNAs were found for sHsp-1A and -1B in early-unicellular stage, increasing four to sevenfold in mature pollen. Nine other genes are up-regulated in unicellular and down-regulated in bicellular pollen; three these genes show stage-specific expression. Western analysis revealed that cytosolic class I and II sHsp are developmentally expressed during all stages of pollen development. Different subsets of cytosolic sHsp genes are expressed in a stage-specific fashion suggesting that certain sHsp genes may play specific roles in early, others during later stages of pollen development. Heat stress results in a relatively weak and incomplete response in pollen: (i) the heat-induced levels of mRNA (excepting sHsp-2B, -3C and -6) are much lower than in leaves, (ii) several sHsp are not detected after heat stress in pollen, although, they are heat-inducibly expressed in leaves. Application of heat stress, cold, and starvation, which induce microspore embryogenesis, modify mRNA levels and the patterns of 2-D separated sHsp, but only heat stress enhances the expression of sHsp in microspores. There is no correlation of the expression of specific sHsp with the potential for microspore embryogenesis. PMID- 15821977 TI - pSAT vectors: a modular series of plasmids for autofluorescent protein tagging and expression of multiple genes in plants. AB - Autofluorescent protein tags represent one of the major and, perhaps, most powerful tools in modern cell biology for visualization of various cellular processes in vivo. In addition, advances in confocal microscopy and the development of autofluorescent proteins with different excitation and emission spectra allowed their simultaneous use for detection of multiple events in the same cell. Nevertheless, while autofluorescent tags are widely used in plant research, the need for a versatile and comprehensive set of vectors specifically designed for fluorescent tagging and transient and stable expression of multiple proteins in plant cells from a single plasmid has not been met by either the industrial or the academic communities. Here, we describe a new modular satellite (SAT) vector system that supports N- and C-terminal fusions to five different autofluorescent tags, EGFP, EYFP, Citrine-YFP, ECFP, and DsRed2. These vectors carry an expanded multiple cloning site that allows easy exchange of the target genes between different autofluorescence tags, and expression of the tagged proteins is controlled by constitutive promoters, which can be easily replaced with virtually any other promoter of interest. In addition, a series of SAT vectors has been adapted for high throughput Gateway recombination cloning. Furthermore, individual expression cassettes can be assembled into Agrobacterium binary plasmids, allowing efficient transient and stable expression of multiple autofluorescently tagged proteins from a single vector following its biolistic delivery or Agrobacterium-mediated genetic transformation. PMID- 15821979 TI - Salicylic acid-inducible Arabidopsis CK2-like activity phosphorylates TGA2. AB - We demonstrate that TGA2, TGA5 and TGA6, and TGA3 to a lesser extent, are phosphorylated by an activity in rabbit reticulocytes. Using deletion and point mutagenesis of TGA2, three amino acid (aa) residues, (11)Ser, (12)Thr and (16)Thr, were found to be critical for efficient phosphorylation by a kinase(s) in rabbit reticulocytes. These three residues also were important for phosphorylation by recombinant human Casein Kinase II (CK2) and by a CK2-like kinase in Arabidopsis leaf extracts. Salicylic acid (SA) treatment enhanced the phosphorylation of recombinant TGA2 in vitro; it also enhanced phosphorylation of a TGA2-GFP fusion protein in vivo. By contrast, in vivo phosphorylation of a TGA2 A-GFP fusion protein, in which the (11)Ser, (12)Thr and (16)Thr residues were mutated to non-phosphorylable alanine, was only poorly if at all stimulated by SA treatment. Mutation of the putative CK2 phosphorylation motif did not affect nuclear localization of TGA2. However, the DNA binding activity of TGA2 was reduced by CK2 treatment, whereas that of TGA2-A was unaffected; TGA2's DNA binding activity after incubation in a rabbit reticulocyte lysate also was substantially lower than that of comparably treated TGA2-A. Taken together, these results suggest that phosphorylation at the putative CK2 phosphorylation site negatively regulates the DNA binding activity of TGA2. Analysis of transgenic Arabidopsis overexpressing TGA2-GFP or TGA2-A-GFP, in the absence of SA treatment, revealed that they accumulated similarly elevated levels of PR-1 gene transcripts. Possible reasons why mutations in the putative CK2 phosphorylation site had little effect on PR-1 induction by TGA2 are discussed. PMID- 15821980 TI - ASYMMETRIC LEAVES2-LIKE1 gene, a member of the AS2/LOB family, controls proximal distal patterning in Arabidopsis petals. AB - The formation and the development of the floral organs require an intercalate expression of organ-specific genes. At the same time, meristem-specific genes are repressed to complete the differentiation of the organs in the floral whorls. In an Arabidopsis activation tagging population, a mutant affected in inflorescence architecture was identified. This gain-of-function mutant, designated downwards siliques1 (dsl1-D), has shorter internodes and the lateral organs such as flowers are bending downwards, similar to the loss-of-function brevipedicellus (bp) mutant. The affected gene in dsl1-D appeared to be ASYMMETRIC LEAVES2-LIKE1 (ASL1)/LATERAL ORGAN BOUNDARIES domain gene 36 (LBD36), which is a member of the ASYMMETRIC LEAVES2 (AS2)/LATERAL ORGAN BOUNDARIES (LOB) domain gene family. Analysis of the loss-of-function mutant asl1/lbd36 did not show morphological aberration. Double mutant analysis of asl1/lbd36 together with as2, the ASL1/LBD36 closest homologue, demonstrates that these two members of the AS2/LOB family act partially redundant to control cell fate determination in Arabidopsis petals. Moreover, molecular analysis revealed that overexpression of ASL1/LBD36 leads to repression of the homeobox gene BP, which supports the model that an antagonistic relationship between ASL/LBD and homeobox members is required for the differentiation of lateral organs. PMID- 15821981 TI - High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome. AB - Proteomic studies have addressed the composition of plant chloroplast ribosomes and 70S ribosomes from the unicellular organism Chlamydomonas reinhardtii But comprehensive characterization of cytoplasmic 80S ribosomes from higher plants has been lacking. We have used two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) to analyse the cytoplasmic 80S ribosomes from the model flowering plant Arabidopsis thaliana. Of the 80 ribosomal protein families predicted to comprise the cytoplasmic 80S ribosome, we have confirmed the presence of 61; specifically, 27 (84%) of the small 40S subunit and 34 (71%) of the large 60S subunit. Nearly half (45%) of the ribosomal proteins identified are represented by two or more distinct spots in the 2-DE gel indicating that these proteins are either post-translationally modified or present as different isoforms. Consistently, MS-based protein identification revealed that at least one-third (34%) of the identified ribosomal protein families showed expression of two or more family members. In addition, we have identified a number of non ribosomal proteins that co-migrate with the plant 80S ribosomes during gradient centrifugation suggesting their possible association with the 80S ribosomes. Among them, RACK1 has recently been proposed to be a ribosome-associated protein that promotes efficient translation in yeast. The study, thus provides the basis for further investigation into the function of the other identified non-ribosomal proteins as well as the biological meaning of the various ribosomal protein isoforms. PMID- 15821984 TI - Characterization of fructose-bisphosphate aldolase regulated by gibberellin in roots of rice seedling. AB - Fructose-bisphosphate aldolase is a glycolytic enzyme whose activity increases in rice roots treated with gibberellin (GA). To investigate the relationship between aldolase and root growth, GA-induced root aldolase was characterized. GA3 promoted an increase in aldolase accumulation when 0.1 microM GA3 was added exogenously to rice roots. Aldolase accumulated abundantly in roots, especially in the apical region. To examine the effect of aldolase function on root growth, transgenic rice plants expressing antisense aldolase were constructed. Root growth of aldolase-antisense transgenic rice was repressed compared with that of the vector control transgenic rice. Although aldolase activity increased by 25% in vector control rice roots treated with 0.1 microM GA3, FBPA activity increased very little by 0.1 microM GA3 treatment in the root of aldolase-antisense transgenic rice. Furthermore, aldolase co-immunoprecipitated with antibodies against vacuolar H+ -ATPase in rice roots. In the root of OsCDPK13-antisense transgenic rice, aldolase did not accumulate even after treatment with GA3. These results suggest that the activation of glycolytic pathway function accelerates root growth and that GA3-induced root aldolase may be modulated through OsCDPK13. Aldolase physically associates with vacuolar H-ATPase in roots and may regulate the vacuolar H-ATPase mediated control of cell elongation that determines root length. PMID- 15821985 TI - Molecular characterization of a WEE1 gene homologue in tomato (Lycopersicon esculentum Mill.). AB - Early fruit development in tomato (Lycopersicon esculentum Mill.) proceeds in two distinct phases of growth that comprise cell division and cell expansion, respectively. In pericarp and the jelly like locular tissue of tomato fruit, the transition between cell division to cell expansion is characterized by the arrest of mitotic activity, numerous rounds of nuclear DNA endoreduplication and the inhibition of Cyclin-Dependent Kinase A (CDKA) activity. To investigate whether the WEE1 kinase may play a role during the endoreduplication process, we isolated and characterized the tomato homologue for WEE1. The LeWEE1 gene consisted of 10 exons with a predicted 510 amino acid-long protein. The accumulation of the corresponding transcripts was associated with mitotically active organs: developing fruits, seeds and roots. Interestingly, LeWEE1was expressed in the jelly like locular tissue concomitant with endoreduplication during fruit development. Using tobacco BY-2 synchronized cells, we showed that the WEE1 gene expression is cell-cycle regulated with a maximum transcript accumulation at S phase. Our data indicate the putative dual contribution of LeWEE1 in the classical cell cycle and the endocycle. PMID- 15821978 TI - Systemic response to aphid infestation by Myzus persicae in the phloem of Apium graveolens. AB - Little is known about the molecular processes involved in the phloem response to aphid feeding. We investigated molecular responses to aphid feeding on celery (Apium graveolenscv. Dulce) plants infested with the aphid Myzus persicae, as a means of identifying changes in phloem function. We used celery as our model species as it is easy to separate the phloem from the surrounding tissues in the petioles of mature leaves of this species. We generated a total of 1187 expressed sequence tags (ESTs), corresponding to 891 non-redundant genes. We analysed these ESTs in silico after cDNA macroarray hybridisation. Aphid feeding led to significant increase in RNA accumulation for 126 different genes. Different patterns of deregulation were observed, including transitory or stable induction 3 or 7 days after infestation. The genes affected belonged to various functional categories and were induced systemically in the phloem after infestation. In particular, genes involved in cell wall modification, water transport, vitamin biosynthesis, photosynthesis, carbon assimilation and nitrogen and carbon mobilisation were up-regulated in the phloem. Further analysis of the response in the phloem or xylem suggested that a component of the response was developed more specifically in the phloem. However, this component was different from the stress responses in the phloem driven by pathogen infection. Our results indicate that the phloem is actively involved in multiple adjustments, recruiting metabolic pathways and in structural changes far from aphid feeding sites. However, they also suggest that the phloem displays specific mechanisms that may not be induced in other tissues. PMID- 15821982 TI - Gene expression analysis by cDNA-AFLP highlights a set of new signaling networks and translational control during seed dormancy breaking in Nicotiana plumbaginifolia. AB - Seed dormancy in Nicotiana plumbaginifolia is characterized by an abscisic acid accumulation linked to a pronounced germination delay. Dormancy can be released by 1 year after-ripening treatment. Using a cDNA-amplified fragment length polymorphism (cDNA-AFLP) approach we compared the gene expression patterns of dormant and after-ripened seeds, air-dry or during one day imbibition and analyzed 15,000 cDNA fragments. Among them 1020 were found to be differentially regulated by dormancy. Of 412 sequenced cDNA fragments, 83 were assigned to a known function by search similarities to public databases. The functional categories of the identified dormancy maintenance and breaking responsive genes, give evidence that after-ripening turns in the air-dry seed to a new developmental program that modulates, at the RNA level, components of translational control, signaling networks, transcriptional control and regulated proteolysis. PMID- 15821986 TI - Ammonium transporter genes in Chlamydomonas: the nitrate-specific regulatory gene Nit2 is involved in Amt1;1 expression. AB - Ammonium transport is a key process in nitrogen metabolism. In the green alga Chlamydomonas, we have characterized molecularly the largest family of ammonium transporters (AMT1) so far described consisting of eight members. CrAmt1 genes have an interesting transcript structure with some very small exons. Differential expression patterns were found for each CrAmt1 gene in response to the nitrogen source by using Real Time PCR. These expression patterns were similar under high and low CO2 atmosphere. CrAmt1;1 expression was characterized in detail. It was repressed in both ammonium and nitrate medium, and strongly expressed in nitrogen free media. Treatment with a Glutamine synthetase inhibitor released partially repression in ammonium and nitrate suggesting that ammonium and its derivatives participate in the observed repressing effects. By studying CrAmt1;1 expression in mutants deficient at different steps of the nitrate assimilation pathway, it has been shown that nitrate has a double negative effect on this gene expression; one related to its reduction to ammonium, and a second one by itself. This second effect of nitrate was dependent on the functionality of the regulatory gene Nit2, specific for nitrate assimilation. Thus, NIT2 would have a dual role on gene expression: the well-known positive one on nitrate assimilation and a novel negative one on Amt1;1 regulation. PMID- 15821988 TI - Ectopic expression of a soybean phytase in developing seeds of Glycine max to improve phosphorus availability. AB - A transgenic approach was used to alter soybean seed phytate content by expressing a soybean phytase gene (GmPhy) during seed development to degrade accumulating phytic acid (IP6). An expression vector containing the soybean phytase cDNA controlled by the seed-specific beta-conglycinin promoter (alpha' subunit) was used to transform embryogenic soybean cultures. Plants from four independent transgenic lines were analyzed for transgene integration and seed IP6 levels. The reduction in IP6 levels in transgenic seeds compared to control 'Jack' soybeans ranged from 12.6 to 24.8 as determined by HPLC. A low copy transformant was propagated to the T4 generation and examined in more detail for phytase expression and enzyme activity during seed development. Expression of phytase mRNA and phytase activity increased during seed development, consistent with the use of an embryo-specific promoter. Ectopic phytase expression during seed development offers potential as an effective strategy for reducing phytate content in soybean seed. PMID- 15821987 TI - Isolation of candidate genes for apomixis in Poa pratensis L. AB - The essential feature of apomixis is that an embryo is formed autonomously by parthenogenesis from an unreduced egg of an embryo sac generated through apomeiosis. The genetic constitution of the offspring is, therefore, usually identical to the maternal parent, a trait of great interest to plant breeders. If apomixis were well understood and harnessed, it could be exploited to indefinitely propagate superior hybrids or specific genotypes bearing complex gene sets. A fundamental contribution to the understanding of the genetic control of the apomictic pathway could be provided by a deep knowledge of molecular mechanisms that regulate the reproductive events. In Poa pratensis the cDNA-AFLP method of mRNA profiling allowed us to visualize a total of 2248 transcript derived fragments and to isolate 179 sequences that differed qualitatively or quantitatively between apomictic and sexual genotypes at the time of flowering when the primary stages of apomixis occur. Three ESTs were chosen for further molecular characterization because of their cDNA-AFLP expression pattern and BLAST information retrieval. The full-lengths of the newly isolated genes were recovered by RACE and their temporal expression patterns were assessed by RT-PCR. Their putative role in cell signaling transduction cascades and trafficking events required during sporogenesis, gametogenesis and embryogenesis in plants is reported and discussed. PMID- 15821989 TI - Promotion of photomorphogenesis by COP1. AB - CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) represses photomorphogenesis in darkness by targeting nuclear-localized transcription factors to proteasome-mediated degradation. Upon light exposure, COP1 migrates to the cytosol allowing photomorphogenesis to proceed but the residual nuclear pool down-regulates light signaling mediated by phytochrome A. Here we show that weak alleles of cop1 exhibit reverse photomorphogenic responses i.e. reduced rather than enhanced cotyledon unfolding under red light compared to darkness. Conversely, COP1 overexpressors which de-etiolate poorly under blue or far-red light, showed enhanced photomorphogenesis under red light. The positive relationship between COP1 and photomorphogenic response required phytochrome B. Thus, genetic manipulation of COP1 levels differentially affects phytochrome A- compared to phytochrome B-mediated responses. We hypothesize that COP1 could be involved in degradation of negative regulators of photomorphogenesis or in transcriptional activation, as observed for some E3 ligases in mammalian development. PMID- 15821990 TI - A root-specific condensing enzyme from Lesquerella fendleri that elongates very long-chain saturated fatty acids. AB - The LfKCS45 gene with a high sequence similarity to known 3-ketoacyl-CoA synthases of the membrane-bound fatty acid elongase was isolated from Lesquerella fendleri. The LfKCS45 gene has a 1464 bp open reading frame without introns, and is predicted to encode a polypeptide of 487 amino acids with an estimated molecular mass of 54.6 kD. High-stringency DNA blot analysis indicated that there were no closely related genes to LfKCS45 in the L. fendleri genome. Analysis of the fatty acid composition of transformed yeast revealed that expression of the LfKCS45 protein results in the synthesis of two novel very-long-chain fatty acids identified as C28:0 and C30:0. LfKCS45 was found to be not active with acyl-CoA substrates C16 to C24 in length. Reverse transcription-PCR experiments showed that the LfKCS45 gene is expressed only in L. fendleri root tips. Histochemical assays for GUS activity in Arabidopsis transformed with the LfKCS45 promoter-GUS fusion construct confirmed this expression pattern and demonstrated that LfKCS45 transcription is restricted to the cells of the lateral root cap. PMID- 15821991 TI - Genome-wide analysis of S-Locus F-box-like genes in Arabidopsis thaliana. AB - The Antirrhinum S-locus F-box gene, AhSLF-S2, has been shown to determine the pollen function of S-RNase-mediated self-incompatibility (SI). Its initial identification led to the discovery of a large family of plant-specific F-box proteins, named the SLF (S-Locus F-box) family, including members from species with or without S-RNase SI system. To investigate the evolution and function of its family members in Arabidopsis, we first identified 92 Arabidopsis F-box proteins related to AhSLF-S2, referred to as AtSFL (S-locus F-box-like) in this report. Phylogenetic analyses with family members from several plant species revealed that they could be classified into five subgroups, and the SLF genes appeared to have had a monophyletic origin. Yeast two-hybrid analyses showed that most AtSFL proteins could interact with one or more ASK (Arabidopsis Skp1-like) proteins, a component of the SCF (Skp1/Cullin or CDC53/F-box) complex, suggesting that AtSFLs may function in the process of ubiquitin/26S proteasome-mediated proteolysis. Transcript analysis found that most of AtSFL genes are expressed ubiquitously and only three of them (AtSFL61, 79 and 85) displayed a tissue specific pattern. In consistent, phenotypic observations for T-DNA insertion lines of 37 AtSFL genes revealed that most of them are functionally redundant, but inactivation of two AtSFL genes (AtSFL 61 and 70) appears to have caused developmental defects in embryo or female gametophyte. Our results show that a diversified expression and functional pattern are associated with AtSFL genes, indicating that they play important roles in various biological processes in Arabidopsis. PMID- 15821992 TI - Gamma carbonic anhydrase like complex interact with plant mitochondrial complex I. AB - We report the identification by two hybrid screens of two novel similar proteins, called Arabidopsis thaliana gamma carbonic anhydrase like1 and 2 (AtgammaCAL1 and AtgammaCAL2), that interact specifically with putative Arabidopsis thaliana gamma Carbonic Anhydrase (AtgammaCA) proteins in plant mitochondria. The interaction region that was located in the N-terminal 150 amino acids of mature AtgammaCA and AtgammaCA like proteins represents a new interaction domain. In vitro experiments indicate that these proteins are imported into mitochondria and are associated with mitochondrial complex I as AtgammaCAs. All plant species analyzed contain both AtgammaCA and AtgammaCAL sequences indicating that these genes were conserved throughout plant evolution. Structural modeling of AtgammaCAL sequences show a deviation of functionally important active site residues with respect to gammaCAs but could form active interfaces in the interaction with AtgammaCAs. We postulate a CA complex tightly associated to plant mitochondrial complex. PMID- 15821993 TI - Functional and regulatory analysis of the Arabidopsis thaliana CAX2 cation transporter. AB - The vacuolar sequestration of metals is an important metal tolerance mechanism in plants. The Arabidopsis thaliana vacuolar transporters CAX1 and CAX2 were originally identified in a Saccharomyces cerevisiae suppression screen as Ca2+/H+ antiporters. CAX2 has a low affinity for Ca2+ but can transport other metals including Mn2+ and Cd2+. Here we demonstrate that unlike cax1 mutants, CAX2 insertional mutants caused no discernable morphological phenotypes or alterations in Ca2+/H+ antiport activity. However, cax2 lines exhibited a reduction in vacuolar Mn2+/H+ antiport and, like cax1 mutants, reduced V-type H+ -ATPase (V ATPase) activity. Analysis of a CAX2 promoter beta-glucoronidase (GUS) reporter gene fusion confirmed that CAX2 was expressed throughout the plant and strongly expressed in flower tissue, vascular tissue and in the apical meristem of young plants. Heterologous expression in yeast identified an N-terminal regulatory region in CAX2, suggesting that Arabidopsis contains multiple cation/H+ antiporters with shared regulatory features. Furthermore, despite significant variations in morphological and biochemical phenotypes, cax1 and cax2 lines both significantly alter V-ATPase activity, hinting at coordinate regulation among transporters driven by H+ gradients and the V-ATPase. PMID- 15821998 TI - A potential role for angiotensin II in obesity induced cardiac hypertrophy and ischaemic/reperfusion injury. AB - BACKGROUND: The mechanisms for obesity induced myocardial remodelling and subsequent mechanical dysfunction are poorly understood. There is good evidence that angiotensin II and TNFalpha have strong growth promoting properties and are elevated with obesity. In addition, these two peptides may interact to exacerbate myocardial ischaemic/reperfusion injury. HYPOTHESIS: Obesity increases systemic and myocardial renin-angiotensin system (RAS) activity and TNFalpha levels and contributes to obesity induced cardiac remodelling and ischaemic/reperfusion injury. METHODS: Male Wistar rats were placed on a standard rat chow diet or cafeteria diet for 16 weeks. Two additional groups of rats received the respective diets and losartan (30 mg/ kg/d) in their drinking water. Hearts were perfused on the isolated working rat heart perfusion system and mechanical function was documented before and after 15 min normothermic total global ischaemia. Blood and myocardial samples were collected for angiotensin II, TNFalpha and NADPH oxidase activity determinations. RESULTS: The rats on the cafeteria diet became obese compared to rats on the standard rat chow (438 +/- 5.9 g vs 393 +/- 7.3 g for control, p < 0.05). Obesity was associated with elevated serum angiotensin II (0.050 +/- 0.015 pmol/ml vs. 0.035 +/- 0.003 pmol/ml, p < 0.05) and TNFalpha levels (42.8 +/- 5.93 pg/ml vs. 13.18 +/- 2.50 pg/ml, p < 0.05), and increased heart to body weight ratios (3.1 +/- 0.04 mg/g vs. 2.8 +/- 0.03 mg/g, p < 0.05). Losartan had no effect on body weight but decreased basal myocardial angiotensin II and TNFAlpha levels as well as heart to body weight ratio in the obese and lean controls (2.5 +/- 0.05 mg/g and 2.6 +/- 0.04 mg/g relative to their controls, p < 0.05). Hearts from obese rats had lower reperfusion aortic outputs (AO) than their concurrent controls (18.42 +/- 1.17 ml/min vs. 27.8 +/- 0.83 ml/min, p < 0.05). Losartan improved aortic output recoveries in obese rats (23.0 +/- 1.71 ml/min, p < 0.05). CONCLUSIONS: Obesity increased serum angiotensin II and TNFalpha levels, blood pressure, and heart weight to body weight ratios. These changes were associated with decreased basal and post-ischaemic myocardial mechanical function. Chronic AT(1) receptor antagonism prevented the adverse changes in heart weight, mechanical function and susceptibility to ischaemic/reperfusion injury. Although current data do not exclude additional mechanisms for obesity induced cardiac remodelling, they suggest that angiotensin II may contribute to obesity induced cardiac remodelling and ischaemic/reperfusion injury. PMID- 15821999 TI - From the microsurgical laboratory to the operating theatre. PMID- 15821994 TI - A hypersensitive response-induced ATPase associated with various cellular activities (AAA) protein from tobacco plants. AB - The hypersensitive response (HR) is one of the most critical defense systems in higher plants. In order to understand its molecular basis, we have screened tobacco genes that are transcriptionally activated during the early stage of the HR by the differential display method. Among six genes initially identified, one was found encoding a 57 kDa polypeptide with 497 amino acids not showing significant similarity to any reported proteins except for the AAA domain (ATPase associated with various cellular activities) spanning over 230 amino acids. The bacterially expressed protein exhibited ATP hydrolysis activity, and a green fluorescent protein-fusion protein localized in the cytoplasm of onion epidermis cells. The protein was subsequently designated as NtAAA1 (Nicotiana tabacum AAA1). NtAAA1 transcripts were induced 6 h after HR onset not only by TMV but also by incompatible Psuedomonas syringae, indicating that NtAAA1 is under the control of the N-gene with a common role in pathogen responses. Expression of NtAAA1 was induced by jasmonic acid and ethylene, but not by salicylic acid (SA). It also occurred at a high level in SA-deficient tobacco plants upon TMV infection. When NtAAA1 was silenced by the RNAi method, accumulation of transcripts for PR-1a significantly increased during the HR. Treatments with SA induced higher expression of PR-1a and acidic PR-2 in RNAi transgenic plants than in wild-type counterparts. These results suggest that NtAAA1 mitigates the SA signaling pathway, and therefore that NtAAA1 modulates the pathogen response of the host plants by adjusting the HR to an appropriate level. PMID- 15822000 TI - Motion repulsion arises from stimulus statistics when analyzed with a clustering algorithm. AB - Motion repulsion is the perceived enlargement of the angle between the directions of motion of two transparently moving patterns. An explanation of this illusion has long been sought for in the neural circuitry of the brain. We show that motion repulsion already arises from the statistical properties of the motion transparency problem when analyzed with a clustering algorithm. PMID- 15822001 TI - Regulation of ion transport via apical purinergic receptors in intact rabbit airway epithelium. AB - We investigated purinergic receptors involved in ion transport regulation in the intact rabbit nasal airway epithelium. Stimulation of apical membrane P2Y receptors with ATP or UTP (200 microM) induced transient increases in short circuit current (Isc) of 13 and 6% followed by sustained inhibitions to 8 and 17% below control level, respectively. Serosal application of nucleotides had no effect. The ATP-induced response appeared to involve additional activation of apical adenosine (P1) and P2X receptors. The inhibitory effect of ATP and UTP on Isc was eliminated by pretreatment with amiloride (100 microM), while the stimulatory effect was potentiated, indicating that ATP and UTP inhibit Na+ and stimulate Cl- current. Ionomycin (1 microM) induced responses similar to UTP and ATP and desensitized the epithelium to the nucleotides, indicating involvement of intracellular Ca2+ (Ca2+ i. Furthermore, ATP, UTP and ionomycin induced 21, 24, and 21% decreases, respectively, in transepithelial conductance. Measurements of unidirectional isotope fluxes showed a 39% decrease in the dominant net Na+ absorption in response to ATP, while the smaller net Cl- secretion increased only insignificantly and unidirectional Cl- fluxes decreased significantly. The results suggest that nucleotides released to the airway surface liquid exert an autocrine regulation of epithelial NaCl absorption mainly by inhibiting the amiloride-sensitive epithelial Na+ channel (ENaC) and paracellular anion conductance via a P2Y receptor-dependent increase in Ca2+ i, while stimulation of Cl- secretion is of minor importance. PMID- 15822002 TI - Effects of levodopa oral bolus on the kinematics of the pointing movements in Parkinson's disease patients. AB - We studied the time-course of a levodopa oral bolus effects on the kinematics of patients affected by a mild akinetic-rigid form of idiopathic Parkinson's disease (PD). Eleven PD patients were evaluated: a) in OFF-state, that is before their first medication or after its withdrawal, b) in ON-state, that is at 1/2, 1, 2, 3, 4, 5, 6, 24, 30 and 48 hours after the administration of 250 mg of levodopa plus 25mg of carbidopa. The main kinematics (i. e.movement time, peak of velocity, peak of acceleration and peak of deceleration) of pointing movements to six target-stimuli placed on the horizontal plane of a table were recorded. Clinical conditions were assessed according to the Motor Examination section of the Unified Parkinson's Disease Rating Scale. The levopoda bolus had stable clinical effects only within the first six hours from its administration. The decline of the clinical response was marked by the changes of peak acceleration whereas other kinematics (i. e. movement time and the peak of velocity) changed also in the late observations (24, 30 and 48 hours after drug intake). The dissociation between the persistent improvement on movement time on peak velocity and the rapid deterioration of levodopa effects on early kinematics (i. e. peak acceleration) could be accounted for by a progressive decline in movement programming. PMID- 15822003 TI - Epilepsy and intracranial calcification of unknown origin. PMID- 15822004 TI - Electrical interaction of mechanosensitive fibroblasts and myocytes in the heart. AB - Fibroblasts in the heart can respond to mechanical deformation of the plasma membrane with characteristic changes of their membrane potential. Membrane depolarization of the fibroblasts occurs during the myocardial contractions and is caused by an influx of cations, mainly of sodium ions, into the cells. Conversely, application of mechanical stretch to the cells, i.e., during diastolic relaxation of the myocardium, will hyperpolarize the membrane potential of the fibroblasts due to reduced sodium entry. Thus, cardiac fibroblasts can function as mechano-electric transducers that are possibly involved in the mechano-electric feedback mechanism of the heart. Mechano-electric feedback refers to the phenomenon, that the cardiac mechanical environment, which depends on the variable filling pressure of the ventricles, modulates the electrical function of the heart. Increased sensitivity of the cardiac fibroblasts to mechanical forces may contribute to the electrical instability and arrhythmic disposition of the heart after myocardial infarction. Novel findings indicate that these processes involve the intercellular transfer of electrical signals between fibroblasts and cardiomyocytes via gap junctions. In this article we will discuss the recent progress in the electrophysiology of cardiac fibroblasts. The main focus will be on the intercellular pathways through which fibroblasts and cardiomyocytes communicate with each other. PMID- 15822006 TI - Brassinosteroid-promoted growth. AB - Brassinosteroids (BRs) are highly potent growth-promoting sterol derivatives. BR deficient or BR-insensitive mutants display dwarfism. Whole plants and excised tissues have been used to analyse the mechanisms involved in BR-promoted growth. BR stimulates cell elongation and cell division, and BR has specific effects on differentiation. Underlying physiological pathways include modification of cell wall properties, effects on carbohydrate assimilation and allocation, and control of aquaporin activities. BR apparently coordinates and integrates diverse processes required for growth, partly via interactions with other phytohormones setting the frame for BR responses. Ultimately, BR-promoted growth is mediated through genomic pathways. Positive regulators of the BR response (such as BZR1 and BES1) and putative downstream components (such as EXO) are involved in the regulation of BR-responsive genes and growth promotion. BR-responsive genes have been identified in several plant species. However, causal links between physiological effects and changes of transcript patterns, for the most part, are still unresolved. This review focuses on physiology and molecular mechanisms underlying BR-promoted growth in the different plant organs. Interactions with other phytohormones are discussed. PMID- 15822007 TI - The inheritance of chilling tolerance in tomato (Lycopersicon spp.). AB - During the past 25 years, chilling tolerance of the cultivated (chilling sensitive) tomato Lycopersicon esculentum and its wild, chilling-tolerant relatives L. peruvianum and L. hirsutum (and, less intensively studied, L. chilense) has been the object of several investigations. The final aim of these studies can be seen in the increase in chilling tolerance of the cultivated genotypes. In this review, we will focus on low-temperature effects on photosynthesis and the inheritance of these traits to the offspring of various breeding attempts. While crossing L. peruvianum (male symbol) to L. esculentum (female symbol) so far has brought the most detailed insight with respect to physiological questions, for practical purposes, e.g., the readily cross ability, crossing programmes with L. hirsutum as pollen donor at present seem to be a promising way to achieve higher chilling-tolerant genotypes of the cultivated tomato. This perspective is due to the progress that has been made with respect to the genetic basis of chilling tolerance of Lycopersicon spp. over the past five years. PMID- 15822008 TI - Parthenocarpic fruit development in tomato. AB - Parthenocarpic fruit development is a very attractive trait for growers and consumers. In tomato, three main sources of facultative parthenocarpy, pat, pat 2, pat-3/pat-4, are known to have potential applications in agriculture. The parthenocarpic fruit development in these lines is triggered by a deregulation of the hormonal balance in some specific tissues. Auxins and gibberellins are considered as the key elements in parthenocarpic fruit development of those lines. An increased level of these hormones in the ovary can substitute for pollination and trigger fruit development. This has opened up genetic engineering approaches for parthenocarpy that have given promising results, both in quality and quantity of seedless fruit production. PMID- 15822009 TI - The distribution of membrane-bound 14-3-3 proteins in organelle-enriched fractions of germinating lily pollen. AB - Proteins of the 14-3-3 family show a broad range of activities in plants, depending on their localisation in different cellular compartments. Different organelle membranes of pollen grains and pollen tubes of Lilium longiflorum Thunb. were separated simultaneously using optimised discontinuous sucrose density centrifugation. The obtained organelle-enriched fractions were identified as vacuolar, Golgi, endoplasmic reticulum and plasma membranes, according to their marker enzyme activities, and were assayed for membrane-bound 14-3-3 proteins by immunodetection. 14-3-3 proteins were detected in the cytoplasm as well as in all obtained organelle fractions but were also released into the extracellular medium. In pollen grains, much more plasma membrane-bound 14-3-3 proteins were detected than in the PM-enriched fraction of pollen tubes, whereas the level of Golgi- and ER-associated 14-3-3 proteins was similar in pollen grains and tubes. This shift in the localisation of membrane-associated 14-3-3 proteins is probably correlated with a change in the major function of 14-3-3 proteins, e.g., perhaps changing from initiating pollen grain germination by activation of the PM H +-ATPase to recruitment of membrane proteins via the secretory pathway during tube elongation. PMID- 15822010 TI - Cytological development and sesquiterpene lactone secretion in capitate glandular trichomes of sunflower. AB - The secretion of sesquiterpene lactones (STL) in capitate glandular trichomes from the anther appendages of Helianthus annuus L. (Asteraceae) was observed by light and fluorescence microscopy and HPLC analysis. Disk flowers within the sunflower capitulum showed the known ontogenetic progression from the centre to the margin. During development of the florets, the trichomes in the anther appendages secreted their metabolites into the subcuticular secretion storage space in front of the two apical cells. All stages of forming the cuticular globe, from the pre-secretory to the post-secretory phase, could be observed microscopically and secretory activity was simultaneously monitored. Six germacrolides and heliangolides of known structure were selected for quantitative analysis. The increase in STL content during extension of the subcuticular space was monitored by HPLC analysis. Thereby, the start and termination of STL biosynthesis was defined in relation to other developmental stages of floret ontogenesis, particularly, the pollen formation. Part of the secreted material showed autofluorescence which could be attributed to a hydroxy-trimethoxy flavone, as determined by NMR and mass spectroscopy. The anther trichomes were cytologically and chemically similar to foliar glandular trichomes of sunflower and represent the multicellular capitate glandular trichome type common to many Asteraceae. The ease with which anther trichomes of H. annuus can be harvested and analyzed suggests that they can provide a valuable model system for investigation of STL and flavonoid metabolism in Asteraceae. PMID- 15822011 TI - Thermal dissipation of light energy is regulated differently and by different mechanisms in lichens and higher plants. AB - Modulated chlorophyll fluorescence was used to compare dissipation of light energy as heat in photosystem II of homoiohydric and poikilohydric photosynthetic organisms which were either hydrated or dehydrated. In hydrated chlorolichens with an alga as the photobiont, fluorescence quenching revealed a dominant mechanism of energy dissipation which was based on a protonation reaction when zeaxanthin was present. CO2 was effective as a weak protonating agent and actinic light was not necessary. In a hydrated cyanobacterial lichen, protonation by CO2 was ineffective to initiate energy dissipation. This was also true for leaves of higher plants. Thus, regulation of zeaxanthin-dependent energy dissipation by protonation was different in leaves and in chlorolichens. A mechanism of energy dissipation different from that based on zeaxanthin became apparent on dehydration of both lichens and leaves. Quenching of maximum or Fm fluorescence increased strongly during dehydration. In lichens, this was also true for so called basal or Fo fluorescence. In contrast to zeaxanthin-dependent quenching, dehydration-induced quenching could not be inhibited by dithiothreitol. Both zeaxanthin-dependent and dehydration-induced quenching cooperated in chlorolichens to increase thermal dissipation of light energy if desiccation occurred in the light. In cyanolichens, which do not possess a zeaxanthin cycle, only desiccation-induced thermal energy dissipation was active in the dry state. Fluorescence emission spectra of chlorolichens revealed stronger desiccation induced suppression of 685-nm fluorescence than of 720-nm fluorescence. In agreement with earlier reports of , fluorescence excitation data showed that desiccation reduced flow of excitation energy from chlorophyll b of the light harvesting complex II to emitting centres more than flow from chlorophyll a of core pigments. The data are discussed in relation to regulation and localization of thermal energy dissipation mechanisms. It is concluded that desiccation induced fluorescence quenching of lichens results from the reversible conversion of energy-conserving to energy-dissipating photosystem II core complexes. PMID- 15822012 TI - Flavanols in somatic cell division and male meiosis of tea (Camellia sinensis) anthers. AB - Young anthers excised from closed tea flower buds ( Camellia sinensis L.) were stained as fresh tissues with p-dimethylaminocinnamaldehyde reagent to localize flavanols associated with nuclei and chromosomes, apart from those flavanols stored in vacuoles. This staining reagent yields a blue colour for flavanols. In the nonsporogenic somatic cells of developing anthers, flavanols were found to be attached to chromosomes at all mitotic stages. Male meiosis started at a bud size of about 3.5 mm in diameter in pollen mother cells which displayed generally more or less pronounced blue nuclei and cytoplasm. The meiotic divisions from prophase I to telophase II were characterized by blue stained nuclei and chromosomes, but within the cytoplasm there was, if any, a random and very poor reaction for flavanols. Metaphase and telophase of meiotic divisions showed maximally condensed chromosomes staining dark blue. Early in telophase II, the cytoplasm was again stained blue; this faded at late tetrad stage. Flavanols of young mitotic and older non-mitotic anthers were determined using high pressure liquid chromatography--chemical reaction detection (HPLC-CRD). Catechin, epicatechin, B2, and epigallocatechin were minor compounds, whereas epicatechin gallate and epigallocatechin gallate were found in higher amounts. The major flavanol compound of the anthers, epicatechin gallate, exhibited a significant affinity to histone sulphate, as shown by UV-VIS spectroscopic titration. PMID- 15822013 TI - Redundancy of stomatal control for the circadian photosynthetic rhythm in Kalanchoe daigremontiana Hamet et Perrier. AB - In continuous light, the Crassulacean acid metabolism plant Kalanchoe daigremontiana Hamet et Perrier has a circadian rhythm of gas exchange with peaks occurring during the subjective night. The rhythm of gas exchange is coupled to a weak, reverse phased rhythm of quantum yield of photosystem II (Phi (PSII)). To test if the rhythm of Phi (PSII) persists in the absence of stomatal control, leaves were coated with a thin layer of translucent silicone grease which prevented CO2 and H2O exchange. In spite of this treatment, the rhythm of Phi (PSII) occurred with close to normal phase timing and with a much larger amplitude than in uncoated leaves. The mechanism underlying the Phi (PSII) rhythm in coated leaves can be explained by a circadian activity of phosphoenolpyruvate carboxylase (PEPC). At peaks of PEPC activity, the small amount of CO2 contained in the coated leaf could have become depleted, preventing the carboxylase activity of Rubisco and causing decreases in electron transport rates (observed as deep troughs of Phi (PSII) at 23-h in LL and at ca. 24-h intervals afterwards). Peaks of Phi (PSII) would be caused by a downregulation of PEPC leading to improved supply of CO2 to Rubisco. Substrate limitation of photochemistry at 23 h (trough of Phi (PSII)) was also suggested by the weak response of ETR in coated leaves to stepwise light enhancement. These results show that photosynthetic rhythmicity in K. daigremontiana is independent of stomatal regulation and may originate in the mesophyll. PMID- 15822014 TI - Environmental constraints on phenology and internal nutrient cycling in the Mediterranean winter-deciduous shrub Amelanchier ovalis Medicus. AB - The functional adjustments of winter-deciduous perennials to Mediterranean conditions have received little attention. The objectives of this study were: (i) to determine whether Amelanchier ovalis, a winter-deciduous shrub of Mediterranean and sub-Mediterranean regions, has nutritional and phenological traits in common with temperate zone deciduous trees and shrubs and (ii) to determine the constraints of Mediterranean environmental conditions on these traits. Over two years, phenology and nitrogen, and phosphorus concentrations were monitored monthly in the crown of A. ovalis. Leaf longevity, survival and nutrient resorption from senescing leaves were used to infer nutrient use efficiency and retention times of nutrients within the crown. In A. ovalis, bud burst was much earlier than in temperate deciduous trees and shrubs. Most vegetative and reproductive growth occurred in spring. Limited phenological development took place during the summer drought period. Unexpectedly, leaf shedding was very gradual, which might be related to water shortages in summer. Leaf longevity, nutrient resorption from senescing leaves, and maximum leaf nutrient concentrations indicated that nutrient retention times were short and nutrient use efficiency was low compared to that found in temperate deciduous plants and co-occurring Mediterranean evergreens. A. ovalis exhibited phenological development appropriate for a Mediterranean climate, although its limited ability to retain nutrients likely restricts the types of sites that it can occupy. PMID- 15822015 TI - Heritable variation in seed sex ratio of the stinging nettle (Urtica dioica). AB - Male and female flowering plants of the dioecious Urtica dioica occur in approximately equal numbers in our study area on the coastal sand dunes of Meijendel. The seed sex ratio (SSR, fraction of males) collected from female plants in the field varied between 0.05 and 0.76, and differed significantly between maternal parents. After one generation of selection for either high or low SSR, female plants produced seed batches with sex ratios as extreme as 0.08 and 0.73. Natural populations of U. dioica harbour considerable genetic variation in SSR. PMID- 15822016 TI - Sexual dimorphism and gynoecium size variation in the andromonoecious shrub Caesalpinia gilliesii. AB - The degree of sexual dimorphism in flowers and inflorescences can be evaluated early in flower development through the study of floral organ size co-variation. In the present work, the gynoecium-androecium size relationship was studied to assess the degree of sexual expression in flowers and inflorescences of the andromonoecious shrub Caesalpinia gilliesii. The co-variation pattern of floral organ sizes was compared between small and large inflorescences, under the hypothesis that inflorescence size reflected differential resource availability. Also, staminate and perfect flowers were collected from three populations and compared on the basis of gynoecium, ovule length, filament length, pollen size and number. The obtained results indicated that staminate and perfect flowers differed only in the gynoecium and ovule length, whereas filament length, pollen size, and number varied across populations. The gynoecium size was smaller and its variability was much higher in staminate than in perfect flowers, as explained by a recent hypothesis about pollinator-mediated gynoecium size selection acting upon perfect flowers. The analysis of the gynoecium-androecium size relationship during flower development, revealed a dissociation of gynoecium growth relative to other floral structures in some buds. Lower gynoecium androecium regression slopes and smaller gynoecia length characterized smaller inflorescences, thus reflecting the fact that sexual expression was more male biased. This trend is in agreement with a differential resource-related response at the inflorescence level, however, post-mating resource allocation and the inclusion of other modular levels may also help us to understand the variation in sexual dimorphism in this species. PMID- 15822017 TI - Reproductive biology of the epiphytic bromeliad Werauhia gladioliflora in a premontane tropical forest. AB - The floral phenology, fruit and seed production, and self-compatibility of Werauhia gladioliflora, an epiphytic bromeliad with a wide distribution, were studied in a premontane forest in the Monteverde area in Costa Rica. The species presents the pollination syndrome of chiropterophily, and it is visited by the small bats Hylonycteris underwoodi and Glossophaga commissarisi (Glossophaginae). The population flowering period extended from October to early December (end of rainy season) and seed dispersal occurred from February to April (dry season). Most plants opened a single flower per night, either every day or at one-day intervals during the flowering period. In natural conditions, the average fruit set amounted to almost half of the potential output, but individual fecundity (number of seeds) remained high. Seed number per fruit and germination capacity after artificial selfing and out-crossing treatments did not differ from natural pollination conditions. Werauhia gladioliflora exhibited high levels of autonomous self-pollination and self-compatibility at the individual and population level, characters associated with the epiphytic habitat. These reproductive traits are also associated with early colonizer species, yet life history traits, such as seed dispersal, seedling establishment success, and growth, are likely to have a major role in determining the presence of this species in the successional vegetation patches scattered over the studied premontane area. PMID- 15822018 TI - Patterns of interaction between Populus Esch5 and Piriformospora indica: a transition from mutualism to antagonism. AB - Piriformospora indica (Sebacinaceae, Basidiomycota) is an axenically cultivable, plant growth promoting root endophyte with a wide host range, including Populus. Rooting of Populus Esch5 explants started within 6 days after transfer to WPM medium. If such plantlets with roots were inoculated with P. indica, there was an increase in root biomass, and the number of 2nd order roots was increased significantly. A totally different observation was recorded when the explants were placed into WPM with pre-grown P. indica. The interaction led to complete blocking of root production and severely inhibited plant growth. Additionally, branched aerial roots appeared which did not penetrate the medium. On contact with the fungal colony or the medium, the ends of the aerial roots became inflated. Prolonged incubation stimulated the fungus to colonize aerial parts of the plant (stem and leaves). Mycelium not only spread on the surface of the aerial parts, but also invaded the cortical tissues inter- and intracellularly. Detached Populus leaves remained vital for 4 - 5 weeks on sterile agar media or on AspM medium with pre-grown P. indica. When the fungus was pre-grown on culture media such as WPM, containing ammonium as the main source of nitrogen, leaves in contact with the cultures turned brownish within 4 - 12 h. Thereafter, the leaves bleached, and about one day later had become whitish. Thus, cultural conditions could alter the behaviour of the fungus drastically: the outcome of the interaction between plant and fungus can be directed from mutualistic to antagonistic, characterized by fungal toxin formation and extension of the colonization to Populus shoots. PMID- 15822019 TI - AFP/beta-HCG secreting CNS germ cell tumors: long-term outcome with respect to initial symptoms and primary tumor resection. Results of the cooperative trial MAKEI 89. AB - PURPOSE: The aim of the present study was to evaluate survival and factors influencing long-term outcome of patients with AFP/beta-HCG secreting (non seminomatous) central nervous system germ cell tumors (secCNSGCT), who were prospectively collected in the cooperative MAKEI (German: maligne Keimzelltumoren) 89 protocol. PATIENTS AND METHODS: Between January 1989 and January 1994, 28 patients with secCNS GCT were registered and treated according to the MAKEI 89 protocol. The protocol recommended, after a clinically or histologically proven diagnosis and cisplatin-based chemotherapy, a resection of residual tumor and craniospinal irradiation (30 Gy) with a tumor boost (20 Gy). RESULTS: The estimated (Kaplan-Meier) event-free survival (EFS) of protocol patients is 0.57 +/- 0.09 (n = 28) and the relapse-free survival (RFS) is 0.67 +/ 0.10 (at five and ten years). With respect to long-term survival, the combination of marked neurological symptoms at diagnosis along with primary tumor resection seem to be the main negative prognostic risk factors (Fisher exact test p < 0.05). CNS dissemination at diagnosis can also be considered as a negative risk factor as 3 of 5 patients with primary dissemination died of the disease. CONCLUSION: Cisplatin-based three agent chemotherapy followed by resection of the residual tumor and craniospinal irradiation (CSI) with tumor boost is a successful and well-tolerated treatment for secCNSGCTs. The possibility of a clinical diagnosis based on MRI and tumor markers together with the use of modern neurosurgical techniques gives us the chance to postpone or even avoid major surgery. This gives an additional chance to reduce acute morbidity and further decrease late effects. PMID- 15822020 TI - Predictive value of neonatal MRI with respect to late MRI findings and clinical outcome. A study in infants with periventricular densities on neonatal ultrasound. AB - PURPOSE: The aim of this study was to correlate hypoxic-ischemic white matter damage on neonatal MRI with MRI appearance and neurological outcome at the age of 1 1/2 years. PATIENTS AND METHODS: A sequential cohort of infants with periventricular densities on neonatal ultrasound was studied with neonatal MRI. Images of 46 infants with a mean gestational age of 31 weeks were obtained at a mean age of 20 days after birth and at 1 1/2 years. To establish agreement between the neonatal and follow-up MRI (general, motor, and visual scores), the weighted Cohen's kappa test was used. To establish the predictive power of neonatal MRI with respect to the neurologic indices at the age of 1 1/2 years, the sensitivity, specificity, and positive and negative predictive values were calculated. RESULTS: There was a moderately good to good agreement between the general, motor, and visual neonatal and follow-up MRI scores: weighted kappa = 0.59 (95% CI: 0.44 - 0.74), 0.82 (95% CI: 0.72 - 0.93), and 0.70 (95% CI: 0.56 - 0.84), respectively. Neonatal MRI scores provided a good prediction of the three neurological outcome measures (developmental delay, cerebral palsy, and cerebral visual impairment): sensitivity, specificity, and predictive values were high, with little difference between the three MRI scores. The 32 patients with (nearly) normal neonatal MRI scores were neurologically (nearly) normal at 1 1/2 years on all three outcome measures, whereas 8 patients with seriously abnormal neonatal MRI scores were neurologically abnormal at 1 1/2 years on all three outcome measures. CONCLUSION: Neonatal MRI is able to predict the precise localization and size of perinatal leukomalacia on follow-up MRI and provides a good prediction of neurological outcome at 1 1/2 years. PMID- 15822021 TI - The first three years of the Swiss Neuropaediatric Stroke Registry (SNPSR): a population-based study of incidence, symptoms and risk factors. AB - We report the results of three years of the population-based, prospective Swiss NeuroPaediatric Stroke Registry (SNPSR) of children (up to 16 years) with childhood arterial ischaemic stroke (AIS1), neonatal stroke (AIS2), or symptomatic sinus venous thrombosis (SVT). Data on risk factors (RF), presentation, diagnostic work-up, localisation, and short-term neurological outcome were collected. 80 children (54 males) have been included, 40 AIS1, 23 AIS2, and 17 SVT. The data presented will be concentrated on AIS. The presentation for AIS1 was hemiparesis in 77% and cerebellar symptoms and seizures in 20%, respectively. AIS2 presented in 83% with seizures and in 38% with abnormality of muscle tone. Two or more RF were detected in 54%, one RF in 35%. The most prominent RF for AIS1 were infections (40%), followed by cardiopathies and coagulopathies (25% each). AIS2 were frequently related to birth problems. Neurological outcomes in AIS1 and AIS2 were moderate/severe in 45 % and 32 %, respectively. The outcome correlated significantly with the size of infarction (p = 0.013) and age at stroke (p = 0.027). The overall mortality was 6%. Paediatric stroke is a multiple risk problem, which leads to important long-term sequelae. PMID- 15822022 TI - Restless legs syndrome and sleep problems in children and adolescents with insulin-dependent diabetes mellitus type 1. AB - The restless legs syndrome (RLS) occurs in adulthood with a prevalence of 5 to 10% and can be associated with diabetes mellitus. The prevalence in childhood, however, is unknown. We asked consecutive children with type 1 (insulin dependent) diabetes mellitus as well as their parents and siblings about RLS according to a standardised questionnaire. Altogether, 46 patients (25 female, 12.0 +/- 3.7 years), 50 siblings (29 female, 12.3 +/- 5.5 years) and 75 parents (41 mothers, 40.4 +/- 5.1 years; 34 fathers, 42.5 +/- 5.3 years; 1.3 % with diabetes mellitus) were included. One patient (2.2%), one sibling (2.0%), and 14 parents (18.7%) were diagnosed as having RLS. Disturbances of sleep initiating, sleep maintenance and daytime tiredness were similar in patients and siblings. There was a significant association of higher HbA1c values (mean 7.7 +/- 2.2%) with sleep initiating problems. The mean dose of international units of insulin/kg body weight/day (0.79 +/- 0.26 IU) was not associated with the presence of RLS or sleep problems. To conclude, there seems to be no association of diabetes mellitus type 1 with RLS in children and adolescents. However, there is a relationship between diabetes and sleep disturbances and an optimally controlled diabetes mellitus might be an important factor for an improved sleep initiation. PMID- 15822023 TI - Neuroborreliosis causing focal cerebral arteriopathy in a child. AB - A 9-year-old girl presented with an acute right-sided hemiparesis. Initially, the clinical presentation and stable vasculopathic abnormalities on MR and conventional angiography were suspicious of a so-called "transient cerebral arteriopathy". Mild but persistent pleocytosis and an elevated CSF IgG index led to an extensive search for infectious and immunological causes of cerebral vasculitis, eventually revealing neuroborreliosis. Although rare, infectious and potentially treatable causes of arterial ischemic stroke should be considered in every child with a documented cerebral arteriopathy. PMID- 15822024 TI - Congenital hydrocephalus as a rare association with ligneous conjunctivitis and type I plasminogen deficiency. AB - Severe type I plasminogen deficiency is the underlying cause of ligneous conjunctivitis (LC). Furthermore, pseudomembranes may also be found on other mucous membranes (gastrointestinal tract, bronchial system, genital tract). In very rare cases, congenital hydrocephalus has been associated with the more severe forms of the disease and may even precede LC. The pathophysiological mechanism is unclear at present. It is advisable to look for plasminogen deficiency in patients with congenital hydrocephalus, because obstruction of ventriculoperitoneal shunts is possible when such a condition is overlooked. Here, we report a case of LC with hydrocephalus. This report reemphasizes the association of LC with hydrocephalus which is not well known. PMID- 15822025 TI - Isolated CNS relapse in neuroblastoma. AB - We report three patients with primary extracerebral neuroblastoma and central nervous system (CNS) relapse. As the survival of children with metastatic neuroblastoma improves with recent advances in treatment, CNS involvement becomes more frequently detected. MR imaging of the brain and spine are strongly indicated in the follow-up of patients with increased risk of developing CNS disease. PMID- 15822026 TI - Bilateral striatal lesion associated with varicella. AB - Bilateral striatal lesion is characterised by a specific clinical syndrome (encephalopathy with rigidity, irritability, variable pyramidal, and extrapyramidal symptoms, speech abnormalities) and symmetrical lesion of the basal ganglia including the caudate nucleus, the putamen, and occasionally other nuclei. We report three cases in whom bilateral striatal lesion developed in association with varicella. Each patient recovered completely and showed no signs of cognitive deficiency, chorea or hyperkinetic syndrome, all of which have been reported as sequelae of BSL associated with other conditions. These cases suggest that bilateral striatal lesion may be an immune-mediated complication of varicella. PMID- 15822032 TI - Intestinal ulcers due to drug-induced perinuclear anti-neutrophilic cytoplasmic antibody-positive vasculitis and antiphospholipid syndrome. PMID- 15822033 TI - Hyperbilirubinemia in the setting of antiviral therapy. AB - Hyperbilirubinemia is a common side effect of antiviral medications. The mechanisms underlying its development are multiple and unique to each therapy. During administration of antiviral medications, the hyperbilirubinemia observed in the absence of liver injury is most frequently manifested by isolated increases in the indirect-reacting fraction. Relevant mechanisms leading to indirect hyperbilirubinemia in this setting include hemolysis, decreased hepatic bilirubin clearance as a result of impairment of bilirubin conjugation, or circumstances in which both processes occur simultaneously. Underlying genetic susceptibilities may potentiate these side effects of antiviral therapy. Conjugated (direct-reacting) hyperbilirubinemia can be a consequence of generalized hepatocellular injury, selective cholestatic defects, biliary obstruction, or, rarely, genetic disorders of bilirubin transport. In the specific setting of antiviral therapy, preexisting liver disease or antiviral hepatotoxicity, such as is encountered with the use of the nucleoside and non nucleoside human immunodeficiency virus reverse transcriptase inhibitors, are the most frequent causes of direct-reacting or mixed direct- and indirect-reacting hyperbilirubinemia. Modification in antiviral drug choice or dose may be required in cases of liver injury or of brisk hemolysis leading to significant anemia. The mild indirect hyperbilirubinemia associated with impairment in conjugation tends to be well tolerated and of little consequence. The decision to continue or discontinue antiviral therapy in the face of hyperbilirubinemia should be made after an assessment of the cause of the elevated bilirubin level and a thorough assessment of the risks and benefits of antiviral therapy. PMID- 15822034 TI - Management of hepatitis C virus in special populations: patient and treatment considerations. AB - The most common chronic blood-borne infection in the United States is caused by hepatitis C virus. An estimated 3.9 million people (1.8%) in the United States have been infected with the hepatitis C virus, excluding certain subpopulations who are at high risk for hepatitis C virus infection. Among these subpopulations are an estimated 255,000 (15%) of prison inmates and 175,000 (22%) of homeless people. Prevalence of hepatitis C virus infection is also high among veterans (6.6% overall and even higher among homeless veterans). The single most important risk factor for hepatitis C virus infection is injection drug use; up to 90% of illicit injection drug users are infected with hepatitis C virus. This review describes the prevalence of hepatitis C virus in special populations and discusses the treatment options for patients with severe disease, transplant recipients, and patients at high risk for infection. Close monitoring and management of therapeutic side effects are required to assist these patients in adhering to therapy. PMID- 15822036 TI - Genomics, genetic epidemiology, and genomic medicine. AB - Medical science is on the threshold of unparalleled progress as a result of the advent of genomics and related disciplines. Human genomics, the study of structure, function, and interactions of all genes in the human genome, promises to improve the diagnosis, treatment, and prevention of disease. This opportunity is the result of the recent completion of the Human Genome Project. It is anticipated that genomics will bring to physicians a powerful means to discover hereditary elements that interact with environmental factors leading to disease. However, the expected transformation toward genomics-based medicine will occur over decades. It will require efforts of many scientists and physicians to begin now to sort out the vast amounts of information in the human genome and translate it to meaningful applications in clinical practice. Meanwhile, practicing physicians and health professionals need to be trained in the principles, applications, and limitations of genomics and genomic medicine. Only then will we be in a position to benefit patients, which is the ultimate goal of accelerating scientific progress in medicine. In this inaugural article, we introduce and discuss concepts, facts, and methods of genomics and genetic epidemiology that will be drawn on in the forthcoming topics of the clinical genomics series. PMID- 15822037 TI - Value of extended recording time with wireless pH monitoring in evaluating gastroesophageal reflux disease. AB - BACKGROUND & AIMS: Extending an ambulatory pH study by using a wireless capsule system could improve diagnosis of gastroesophageal reflux disease in patients with symptoms potentially related to acid reflux events. We determined the relative value of a 2-day versus a 1-day study in detection of abnormal esophageal acid exposure and reflux symptom associations. METHODS: Subjects who had undergone pH monitoring with the wireless system for symptom evaluation and who had at least 1 typical or atypical symptom during monitoring were studied (n = 157). Acid exposure time, symptom index, and a measure of reflux-associated symptom probability were calculated after 1 day of recording and compared to final results from analysis of 2 days of data. RESULTS: Extending recording time increased the number of subjects recording symptoms by 6.8% and doubled the number of symptoms available for association with acid reflux events. Other advantages were greater in subjects off antireflux therapy, wherein lengthening recording time increased the proportion of subjects with abnormal acid exposure time by 12.4% and significantly increased the likelihood of a positive reflux symptom relationship in subjects with atypical symptoms. Accuracy of a 1-day study ranged from 0.87-0.88 for acid exposure time to as low as 0.72-0.87 for the probability test, the most sensitive of the symptom association tests. CONCLUSIONS: Extending recording time to 2 days with the wireless pH monitoring system increases the likelihood of detecting reflux disease in patients undergoing symptom evaluation. Benefits are most evident for subjects with atypical symptoms and for establishing reflux symptom associations by probability testing. PMID- 15822038 TI - Permeability, zonulin production, and enteropathy in dermatitis herpetiformis. AB - BACKGROUND & AIMS: Dermatitis herpetiformis (DH) is characterized by variable degrees of enteropathy and increased intestinal permeability. Zonulin, a regulator of tight junctions, seems to play a key role in the altered intestinal permeability that characterizes the early phase of celiac disease. Our aim was to assess both intestinal permeability and serum zonulin levels in a group of patients with DH having variable grades of enteropathy. METHODS: We studied 18 DH patients diagnosed on the basis of characteristic immunoglobulin (Ig)A granular deposits in the dermal papillae of noninvolved skin. Results were compared with those of classic celiac patients, patients with linear IgA dermatosis, and healthy controls. RESULTS: According to Marsh's classification, 5 patients had no evidence of enteropathy (type 0), 4 patients had type II, 2 patients had type IIIb damage, and 7 patients had a more severe lesion (type IIIc). Intestinal permeability (lactulose/mannitol ratio [lac/man]) was abnormal in all patients with DH. Patients with more severe enteropathy had significantly greater permeability ( P < .05). The serum zonulin concentration (enzyme-linked immunosorbent assay) for patients with DH was 2.1 +/- .3 ng/mg with 14 of 16 (87.5%) patients having abnormally increased values. In contrast, patients with linear IgA dermatosis had normal histology, normal intestinal permeability, and negative celiac serology. CONCLUSIONS: Increased intestinal permeability and zonulin up-regulation are common and concomitant findings among patients with DH, likely involved in pathogenesis. Increased permeability can be observed even in patients with no evidence of histologic damage in biopsy specimens. Patients with linear IgA dermatosis appear to be a distinct population with no evidence of gluten sensitivity. PMID- 15822039 TI - Rates of functional bowel disorders among Israeli Bedouins in rural areas compared with those who moved to permanent towns. AB - BACKGROUND & AIMS: Half of Israeli Bedouin society has undergone a transition from nomadic existence to permanent towns, causing cultural and social upheaval. The aim was to compare rates of irritable bowel syndrome (IBS) and functional bowel disorders (FBDs) between Israeli Bedouins still living under rural conditions with those in permanent towns. METHODS: Interviews were conducted in Arabic by trained Bedouin interviewers at 8 Bedouin clinics. The same interviewers surveyed the 2 sectors under identical sampling and interviewing conditions at the same time. FBDs were diagnosed by Rome II criteria. RESULTS: One thousand seven hundred fifty-five Bedouins participated, 1018 from permanent towns and 737 from rural areas. Sixty percent were female (58.2% for rural and 62.0% for towns). The mean age was 39.1 +/- 14.1 years (39.0 +/- 14.3 years for towns, 39.2 +/- 13.9 years for rural; P = NS). The mean level of education was 4.3 +/- 5.4 years (4.6 +/- 5.6 years for towns, 3.7 +/- 5.2 years for rural; P < .0001). IBS was diagnosed in 9.4% of town and 5.8% of rural Bedouins ( P < .01). In contrast, rural Bedouins had significantly higher rates of functional abdominal bloating (7.9% vs 2.8%, P < .0001) and a marginally higher rate of functional constipation. Bedouins living in towns attributed their gastrointestinal symptoms to stress more than rural Bedouins did ( P < .05). Stress and poor global feeling of well-being were significant contributors for IBS in logistic regression models for both sectors. CONCLUSIONS: Bedouins living in permanent towns have significantly higher rates of IBS than rural Bedouins. Although these findings might be associated with the stressful social upheaval that they have undergone, further study is needed to substantiate this point. PMID- 15822040 TI - Abnormalities of 5-hydroxytryptamine metabolism in irritable bowel syndrome. AB - BACKGROUND & AIMS: 5-hydroxytryptamine-3 (5-HT 3 ) receptor antagonists improve symptoms in patients with diarrhea-predominant irritable bowel syndrome (D-IBS), 5-HT 4 agonists help those with constipation-predominant IBS (C-IBS). These data suggest excess or deficiency in 5-HT in D-IBS or C-IBS, respectively. Mucosal 5 HT-containing enterochromaffin cells (EC) are increased in postinfectious IBS (PI IBS). Our aim was to define the postprandial release of 5-HT in PI-IBS and C-IBS patients and to relate this to mucosal 5-HT turnover. METHODS: Fifteen PI-IBS patients with diarrhea-predominant symptoms, 15 C-IBS patients, and 15 healthy controls underwent serial (platelet-poor) plasma 5-HT measurement for 3 hours after a standard 520-kcal meal. Rectal biopsy specimens were assayed for 5-HT and its metabolite 5-hydroxindoleacetic acid (5-HIAA). Colonic transit was measured using radio-opaque markers. RESULTS: Colonic transit was prolonged in C-IBS patients (mean +/- SEM) (49.4 +/- 3.8 h) compared with PI-IBS (26.7 +/- 4.5) and control patients (34.1 +/- 4.5) ( P < .02). Release of 5-HT assessed by area under the curve (AUC) of platelet-poor plasma 5-HT from 0 to 180 minutes postprandially was significantly lower in C-IBS patients (2593 +/- 309 mmol/L . min) compared with P-IBS (5623 +/- 721) and control patients (4822 +/- 598) ( P < .001). PI-IBS patients showed significantly higher peak postprandial plasma 5-HT values (median, range) (71.7, 43.4-125.3) ng/L compared with C-IBS patients (31.2, 15.2-40.5) and control patients (43.6, 26.7-50.1) ( P < .01). Mucosal 5-HT turnover as assessed by mucosal 5-HIAA/5-HT ratio was decreased in both C-IBS and PI-IBS patients, .14 (.01-.6) and .21 (.02-2.5), respectively, compared with control patients 1.12 (.17-3.1) ( P < .002). CONCLUSIONS: C-IBS patients show impaired postprandial 5-HT release whereas PI-IBS patients have higher peak levels, abnormalities that may be related to their different symptoms. PMID- 15822041 TI - An oral supplement enriched with fish oil, soluble fiber, and antioxidants for corticosteroid sparing in ulcerative colitis: a randomized, controlled trial. AB - BACKGROUND & AIMS: N-3 fatty acids from fish oil, antioxidants, and short-chain fatty acids (SCFAs) produced during the fermentation of soluble fiber may attenuate inflammation associated with ulcerative colitis (UC). We assessed the efficacy of a nutritionally balanced oral supplement enriched with fish oil, fructooligosaccharides, gum arabic, vitamin E, vitamin C, and selenium on disease activity and medication use in adults with mild to moderate UC. METHODS: A total of 121 patients with UC and a disease activity index (DAI) from 3-9 on a 12-point scale were block randomized for extent of disease and smoking status. In addition to their usual diet, patients consumed 18 oz of the oral supplement or a carbohydrate-based placebo formula each day for 6 months. Clinical and histologic responses were assessed at 3 and 6 months or at the final visit. A change in average prednisone use between groups was tested by using a linear mixed-effects model. RESULTS: Eighty-six patients completed the study. Baseline characteristics were not different between groups except for a higher total DAI score in the oral supplement group (7.3 +/- 1.3; n = 36) compared with the placebo group (6.2 +/- 2.0; n = 50) ( P < .05). Both groups showed significant and similar degree of improvement at 6 months in DAI (-2.5 for oral supplement and -2.8 for placebo) and histologic index (-1.9 for oral supplement vs. -2.0 for placebo). Both intent to-treat and completed patients given oral supplement had a significantly greater rate of decrease in the dose of prednisone required to control clinical symptoms over 6 months as compared with the placebo group ( P < .001). CONCLUSIONS: The improvement in clinical response combined with a decreased requirement for corticosteroids suggest that this enriched oral supplement can be a useful adjuvant therapy in patients with UC. PMID- 15822042 TI - Is desmoplasia a protective factor for survival in patients with colorectal carcinoma? AB - BACKGROUND & AIMS: The role of desmoplasia in colorectal carcinoma progression is unclear and the presence of collagen stroma may represent a barrier against cancer diffusion and vascular invasion or a stroma to build up and support the tumor. The aim of this study was to evaluate the effect of desmoplastic response on long-term survival of patients who underwent radical resection for colorectal carcinoma. METHODS: The study included 429 patients who underwent radical colorectal resection for cancer with a median follow-up period of 72.8 months. RESULTS: At univariate analysis significant associations were observed between desmoplasia and histologic type, parietal infiltration, growth pattern, and staging. No associations were found between desmoplasia and the other clinical and histologic parameters. The multivariate analysis stratified for tumor stage revealed that the factor showing the most favorable influence on time to death was desmoplasia. The presence of desmoplasia was likely to decrease the failure rate to a third of the rate experienced by patients without desmoplasia. Parietal infiltration was associated with an increased risk for a shortened time to death. CONCLUSIONS: Our results favor the view that desmoplasia is a protective factor for survival in patients with colorectal carcinoma. This finding is consistent with the hypothesis that desmoplasia may prevent cancer invasiveness by building a barrier against tumor diffusion. PMID- 15822043 TI - Ulinastatin for pancreatitis after endoscopic retrograde cholangiopancreatography: a randomized, controlled trial. AB - BACKGROUND & AIMS: Pancreatitis remains the major complication of endoscopic retrograde cholangiopancreatography (ERCP), and hyperenzymemia after ERCP is common. Because ulinastatin, a protease inhibitor, has proved effective in the treatment of acute pancreatitis, the aim of this study was to assess the efficacy of ulinastatin for the prevention of post-ERCP pancreatitis and hyperenzymemia. METHODS: In a multicenter, randomized, double-blind, placebo-controlled trial, patients undergoing a first ERCP were randomized to receive ulinastatin (150,000 U) or placebo by intravenous infusion for 10 minutes starting immediately before ERCP. All patients were hospitalized at least 24 hours after ERCP for evaluation of clinical symptoms. Serum pancreatic enzyme levels were measured before and at 4 and 18 hours after ERCP. The primary end point was the incidence of post-ERCP pancreatitis and the secondary objective was the occurrence of hyperenzymemia. RESULTS: A total of 406 patients were enrolled (204 in the ulinastatin group and 202 in the placebo group). There were no differences between the 2 groups regarding baseline characteristics, details of fluoroscopic findings, or endoscopic procedure. The incidence of hyperenzymemia was significantly lower in the ulinastatin group than in the placebo group (amylase, P = .011; lipase, P = .008). Six patients in the ulinastatin group and 15 patients in the placebo group developed pancreatitis (2.9% vs. 7.4%, P = .041). There was no case of severe pancreatitis in either group. Patients who received ulinastatin did not present any side effects related to the medication. CONCLUSIONS: Prophylactic short-term administration of ulinastatin decreases the incidence of pancreatitis and hyperenzymemia after ERCP. PMID- 15822044 TI - Hepatic stellate cells and fibrosis progression in patients with nonalcoholic fatty liver disease. AB - BACKGROUND & AIMS: Many patients with nonalcoholic fatty liver disease (NAFLD) have a benign clinical course, but a subgroup of patients progress to advanced fibrosis and cirrhosis. However, there are no available clinical tools to predict fibrosis progression in this population. Activated hepatic stellate cells (HSCs) are the source of collagen deposition in the liver. We aimed at determining whether an HSC activation score predicts fibrosis progression in NAFLD patients. METHODS: The cohort consisted of 39 untreated patients with NAFLD with paired liver biopsies performed 5-59 months apart (mean, 22 months). Patients were divided into 2 groups on the basis of whether fibrosis progression was noted on their second liver biopsy. Liver tissue was immunostained for alpha-smooth muscle actin, and the HSC score was determined independently by 2 pathologists in the NAFLD population and in control subjects without liver disease. RESULTS: The HSC activation score was significantly increased in patients with fibrosis progression versus patients in whom no fibrosis progression was observed (4.8 +/- 0.5 vs 1.8 +/- 0.6, respectively; P < .001). The HSC score was accurate in predicting fibrosis progression, with a positive predictive value of 90%, specificity of 94%, and an area under the receiver operating characteristic curve of 0.82. However, the negative predictive value and sensitivity were 56% and 41%, respectively. The inter-pathologist agreement for the HSC score was excellent (kappa coefficient, 0.95). CONCLUSIONS: These findings suggest that the HSC activation score is a suitable clinical tool to determine the risk of fibrosis progression in patients with NAFLD. PMID- 15822046 TI - Severe irritable bowel and functional abdominal pain syndromes: managing the patient and health care costs. PMID- 15822045 TI - Factors associated with advanced liver disease in adults with alpha1-antitrypsin deficiency. AB - BACKGROUND & AIMS: Alpha 1 -antitrypsin deficiency (AAT) is an autosomal recessive disease that affects 1 in 2500 persons and might lead to cirrhosis. Our study aim was to characterize the liver disease in AAT and identify factors associated with advanced liver disease. METHODS: A cohort of the Alpha-1 Foundation Registry who reported liver disease was surveyed with a liver disease questionnaire to obtain information related to liver disease, liver transplantation, and AAT phenotype. RESULTS: One hundred sixty-five of the 2175 participants in the registry reported a history of jaundice or liver disease, and 139 (84.2%) completed the questionnaire. Of these, 71.3% were PiZZ, 18.0% were PiMZ, and 5.7% did not know their phenotype. Analysis of 104 participants with a known age of diagnosis included 30 participants diagnosed with liver disease before 18 years, of whom 15 had advanced liver disease defined as liver transplantation or listed for liver transplantation. No differences in age, age at diagnosis, gender, race, phenotype, or infant jaundice were identified. Seventy-four participants were diagnosed after age 18 years, of whom 25 had advanced liver disease. In this group, advanced liver disease was associated with male gender ( P = .006) and a greater mean body mass index ( P = .01), but not with race, Pi phenotype, infant jaundice, diabetes, or hypercholesterolemia. Viral hepatitis was more frequently reported in the nontransplant group (34.7% vs 8.0%, P = .01), and the mean daily alcohol use was significantly greater in this group ( P = .04). CONCLUSIONS: Our results suggest that male gender and obesity but not alcohol or viral hepatitis predispose to advanced liver disease in adults with AAT. PMID- 15822048 TI - Guest editorial: diabetic kidney disease. PMID- 15822049 TI - Diabetic nephropathy: of mice and men. AB - Accumulating evidence supports intrinsic genetic susceptibility as an important variable in the progression of diabetic nephropathy in people. Mice provide an experimental platform of unparalleled power for dissecting the genetics of mammalian diseases; however, phenotypic analysis of diabetic mice lags behind that already established for humans. Standardized benchmarks of hyperglycemia, albuminuria, and measurements of renal failure remain to be developed for different inbred strains of mice. The most glaring deficiency has been the lack of a diabetic mouse model that develops progressively worsening renal insufficiency, the sine qua non of diabetic nephropathy in humans. Differences in susceptibility of these inbred strains to complications of diabetes mellitus provide a possible avenue to dissect the genetic basis of diabetic nephropathy; however, the identification of those strains and/or mutants most susceptible to renal injury from diabetes mellitus is lacking. Identification of a mouse model that faithfully mirrors the pathogenesis of DN in humans will undoubtedly facilitate the development of new diagnostic and therapeutic interventions. PMID- 15822050 TI - Role of oxidative stress in diabetic nephropathy. AB - Accumulating research suggests that oxidative stress is a significant contributor to the pathogenesis of diabetic nephropathy. The normal kidney generates a substantial amount of oxidative stress because of its high metabolic activity that is balanced by an extensive antioxidant system. However, in pathologic states such as hyperglycemia, nitroso-oxidant balance shifts toward a pro-oxidant state that accelerates tissue and vascular injury. This oxidative damage progresses concomitant with worsening glucose metabolism, vascular dysfunction, and kidney disease. Accordingly, strategies to reduce oxidative stress in diabetes mellitus may exert favorable effects on the progression of diabetic nephropathy. PMID- 15822051 TI - Evidence for different susceptibility genes for proteinuria and ESRD in type 2 diabetes. AB - Proteinuria and impaired kidney function are 2 major traits of diabetic nephropathy that aggregate (are heritable) in families of diabetic individuals. Although both traits are heritable, they are not genetically correlated. These findings not only support the hypothesis that the development of diabetic nephropathy consists of 2 distinct disease processes (ie, increasing proteinuria and declining kidney function) but also strongly justify searches for the putative genes that separately determine variation in these processes. These searches have used both genome-wide scans and candidate-gene approaches. By use of genome-scan approaches, several research groups have identified genetic regions on chromosomes 7q, 18q, and 22q that harbor genes that determine either variation in urinary albumin excretion or susceptibility to proteinuria in families who have type 2 diabetes. The evidence for linkage in these 3 genetic regions was suggestive or strong, but, except for 7q, the regions did not overlap across studies. Two genome scans performed in families who have type 2 diabetes identified genetic regions on chromosome 3q, 6q, 7p, and 18q that harbor susceptibility genes that determine variation in glomerular filtration rate or susceptibility to the development of end-stage renal disease (ESRD). The region on 7p overlapped in both studies. Optimism is growing that a positional cloning approach applied to these putative genetic regions will lead to the isolation of the susceptibility genes for proteinuria and ESRD. Meanwhile, significant efforts that make use of the candidate-gene approach have been directed to the search for susceptibility genes for diabetic nephropathy. Unfortunately, positive findings have not been consistent. PMID- 15822052 TI - Detection of urinary albumin. AB - Microalbuminuria is an important clinical marker in patients with diabetes and cardiovascular disease. The concentration of albumin in urine has traditionally been measured by semiquantitative dipsticks or by various quantitative immunochemical methods such as immunonephelometry, immunoturbidimetry, and radioimmunoassay. However, until recently, urinary albumin not reabsorbed by proximal tubular cells was assumed to be excreted intact. Studies have now revealed that the nature of urinary albumin is complex and is excreted as a mixture of intact albumin, albumin-derived peptides that are not detected by routine dipstick and antibody-based tests, and a species of intact albumin (immunounreactive albumin), also not detected by dipstick and antibody-based tests. A new test, Accumin, based on high-performance liquid chromatography analysis, is able to detect all the immunoreactive intact albumin and immunounreactive intact albumin (total intact albumin) in urine. The advantage in the use of Accumin over a conventional dipstick test or antibody-based laboratory method for detecting microalbuminuria is that false negatives are reduced and a relatively earlier diagnosis of incipient kidney disease can be achieved. The introduction of Accumin has, therefore, highlighted the need for a global standard in the detection and measurement of microalbuminuria. By detecting all of the immunoreactive and immunounreactive intact albumin in urine, Accumin has virtually invalidated the use of dye and immunologically-based dipstick tests and immunologically-based laboratory methods in screening for microalbuminuria in diabetic patients and in identifying microalbuminuria as a risk factor for cardiovascular disease. PMID- 15822053 TI - Tubular changes in early diabetic nephropathy. AB - Far from being bystanders in diabetic nephropathy, changes in the proximal tubule are important for the development of progressive diabetic kidney disease. The proximal tubule is uniquely susceptible to a variety of metabolic and hemodynamic factors associated with diabetes. Renal function and prognosis correlate better with structural lesions in the tubuli and cortical interstitium than with classical glomerular changes of diabetic nephropathy. The proximal tubules show a variety of poorly characterized changes, which have led to the notion that tubular damage represents a "final common pathway" for proteinuric renal injury. However, tubular hypertrophy, reduced organic ion transport, and other tubular changes reviewed in this paper, are already apparent before the onset of proteinuria in diabetes. Indeed, increased tubuloglomerular feedback and defective uptake and lysosomal processing may independently contribute to hyperfiltration and urinary protein loss, respectively. This finding does not mean that glomerular or vascular dysfunction do not contribute to progressive nephropathy. However, although subdividing the nephron for the purposes of analysis and scientific discovery may be useful, the interactions between tubule, glomerulus, and interstitium are likely key to the understanding of complex disorders such as diabetic nephropathy. From this "holonephric" point of view, an understanding of the changes in the diabetic tubule forms an important component to the understanding of kidney disease in diabetes. PMID- 15822054 TI - Effects of statin therapy on the progression of chronic kidney disease. AB - Statins are lipid-lowering agents that specifically, competitively, and reversibly inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the enzyme that catalyzes the conversion of HMG-CoA to mevalonic acid, the rate limiting step in the formation of cholesterol. A large body of evidence from numerous, well-controlled, randomized trials demonstrates that statins significantly reduce fatal and nonfatal cardiovascular events in the general population. Cardiovascular benefits of statins have been conventionally attributed to reduction in levels of low-density lipoprotein cholesterol. More recently, subanalyses of large clinical trials suggest that statins may also prove beneficial in ameliorating the progression of kidney disease through their cholesterol-dependent and/or cholesterol-independent (pleiotropic) effects. This review focuses on the role of statin therapy in the progression of chronic kidney disease, the published trials that study the effect of antilipidemic agents on nephropathy, and the emerging pleiotropic effects of statins on the kidneys. PMID- 15822055 TI - Communicating prognosis in the dialysis consent process: a patient-centered, guideline-supported approach. AB - Recent guidelines recommend shared decision making between patients and nephrologists as the model for dialysis decision making. A key component of this shared decision making is obtaining informed consent. As part of this process, nephrologists have an obligation to inform patients with chronic kidney disease of their prognosis. Ideally, patients themselves should be involved in the decision-making process; however, some patients will not possess decision-making capacity, and others may be unwilling to participate. Determining what to tell patients about prognosis requires tailoring the conversation to the individual patient's preferences. Conversations about prognosis need to occur in a timely fashion so that patients have the opportunity to consider options and make decisions before dialysis is inevitable. Communication strategies are available to assist nephrologists in breaking the bad news of the need for dialysis and its associated burdens. The approach described in this article should help nephrologists discuss prognosis with their patients in a way that is patient centered and in accordance with clinical practice guideline recommendations. PMID- 15822056 TI - Predicting the development of diabetic nephropathy and its progression. AB - Diabetes remains the number one cause of end-stage renal disease worldwide. Only about one third of diabetic patients develop nephropathy, and the risk appears to be, in part, genetically determined. In this article, we review clinical and genetic markers for the development and progression of diabetic nephropathy. Microalbuminuria remains the best available predictor of the subsequent development of nephropathy, even though in recent years it has become clear that less than 50% of individuals with type 1 diabetes progress to overt proteinuria over a period of less than 10 years. It is of great interest for early recognition of risk of nephropathy that small elevations in nighttime blood pressure predict microalbuminuria in type 1 diabetes. Genetic markers for diabetic nephropathy have not been conclusively identified. The occurrence of renal events in diabetic patients, however, appears to be influenced by the angiotensin-converting enzyme (ACE) genotype, with a dominant deleterious effect of the D allele (D/D or I/D) versus I/I genotype. Some patients with the DD genotype also appear less susceptible to the renoprotective effects of conventional doses of ACE inhibitors, suggesting that ACE genotyping might be useful in selecting those patients that could benefit from higher doses of ACE inhibitors and more aggressive treatment to prevent or delay disease progression. PMID- 15822057 TI - The next generation of diabetic nephropathy therapies: an update. AB - Although treatments for diabetic kidney disease are available, many patients still have progressive disease. More effective therapies are urgently needed. Novel agents currently under evaluation in clinical trials are described in this review. Sulodexide, a mixture of three glycosaminoglycans, appears to prevent diabetic nephropathy in experimental models by ameliorating abnormalities in the glomerular basement membrane and mesangial matrix. Pyridoxamine is an inhibitor of advanced glycation end-product (AGE) formation derived from vitamin B(6). Alagebrium is an AGE cross-link breaker. AGEs injure the kidneys and other vascular targets by mechanisms such as oxidative stress, inflammation, and protein cross-linking, among others. By inhibiting AGE formation or breaking AGE cross-links, experimental models have demonstrated kidney protection. Ruboxistaurin is an inhibitor of protein kinase C beta (PKC-beta), a mediator of signal transduction that leads to cell growth, fibrosis, and tissue injury. In diabetes, PKC-beta is up-regulated and activated in the kidney. Ruboxistaurin prevents diabetic kidney disease in animal models. These agents have appeared promising (by reduction of albuminuria and preservation of kidney function) in phase II studies. To determine whether clinical outcomes (mortality, renal, and cardiovascular events) are improved beyond the current standard of care, phase III trials are planned. PMID- 15822058 TI - A history of diabetes mellitus or how a disease of the kidneys evolved into a kidney disease. AB - Diabetes mellitus has a long history during which it was considered to be a disease of the kidneys well into the middle of the 19th century. Recognized in antiquity from its excessive urine output and described as a disease of the urinary tract, its clinical features and fatal outcome were quite accurately recorded by the 1st century ad . Galen (129-200) described it as a disease specific to the kidneys because of a weakness in their retentive faculties. The sweet taste of diabetic urine, which is described in ancient Indian texts and noted by Avicenna (980-1037) and Morgagni (1635-1683), was attributed to the passage of absorbed water and nutrients unchanged into the urine. In 1674, Thomas Willis (1621-1675) first differentiated diabetes from other causes of polyuria by the sweet taste (quasi melle) of diabetic urine and suggested that the sweetness first appears in the blood. A century later, Matthew Dobson (1732-1784) showed that the urine sweetness was because of sugar and was preceded and accompanied by sugar in the blood. Although diabetes then came to be ascribed to increased sugar in the blood, the presence of sugar in the urine continued to be attributed to the decreased retentive properties of the kidneys. The experimental production of diabetes in pancreatectomized dogs that could be reversed by subcutaneous pancreatic transplantation in 1889, and ultimate isolation of insulin in 1922 clearly established diabetes as an endocrine disease. The stage of diabetes as a disease of the kidneys was now over but that of diabetes as a cause of kidney disease was yet to come. Diabetes as a cause of end stage kidney disease was first described in 1936 and extensively documented shortly thereafter, whereas the evidence of its increasing prevalence as a cause of chronic kidney disease continues to accrue. PMID- 15822060 TI - The goals of comprehensive and integrated disease state management for diabetic kidney-disease patients. AB - Diabetes, cardiovascular disease, and chronic kidney disease present many challenges to clinicians as separate disease entities. Management and optimization of care of diabetic kidney-disease patients requires a proactive and integrated approach for all 3 disease states. To optimize the assessment, monitoring, and intervention in this population, comprehensive and integrated disease state management medical care plans must be established. Medical nutrition therapy is another key component that must be coordinated with these disease-state management medical care plans. Coordination of integrated disease state management and medical nutrition therapy for diabetes, cardiovascular disease, and chronic kidney disease will help to improve patient outcomes and allow for a proactive approach to the identification, prevention, and management of potential disease-state complications. Several programs and guidelines have been established to accomplish these goals: the K/DOQI Clinical Practice Guidelines, the American Diabetes Association Standards of Medical Care for Diabetes (developed by the American Diabetes Association, the National Institute of Diabetes and Digestive Kidney Diseases, and the Centers for Disease Control Diabetes Foundation), the National Diabetes Education Program, and the National Cholesterol Education Program. The multidisciplinary medical care team assumes an integral role in the success and implementation of this integrated approach as well as the empowerment of the patient in their own care. PMID- 15822059 TI - Pediatric aspects of diabetic kidney disease. AB - Type 1 diabetes mellitus (DM1) commonly occurs in childhood, although many pediatric centers are now seeing more cases of type 2 diabetes (DM2). Kidney failure caused by either type of diabetes is uncommon during childhood, but these years of hyperglycemia contribute to long-term complications. All children with diabetes warrant screening of glomerular filtration rate, blood pressure, and urine albumin excretion. Screening should begin after 5 years of DM1 or at puberty. A similar screening strategy should start at the time of diagnosis of DM2. Atypical features such as dipstick positive proteinuria or active urine sediment may warrant referral to a nephrologist for evaluation, including biopsy. The first line of treatment in either form of diabetes is achieving the best glycemic control possible. Patients developing microalbuminuria or hypertension should receive antiangiotensin II drugs. Adult studies suggest blood pressure goals should be lower in diabetes than in the general population. Although direct evidence is not yet available in children, achieving blood pressure below the 90th percentile for age, height, and gender seems prudent. Longitudinal studies and new screening tests may allow detection of susceptible children earlier in the course of DM1 or DM2, perhaps allowing prevention of diabetic kidney disease. PMID- 15822064 TI - Capillary electrophoresis-inductively coupled plasma-mass spectrometry: a report on technical principles and problem solutions, potential, and limitations of this technology as well as on examples of application. AB - This paper summarizes some basic principles of capillary electrophoresis (CE), inductively coupled plasma-mass spectrometry (ICP-MS), and coupling of both devices. Especially the interfacing is described in detail. A special focus is drawn to various interface developments reported in literature and technical problems, i.e., requirements to the interface setup and respective solutions. Nowadays, typically sheath flow-based interfaces are used. The sheath flow fulfills two requirements of hyphenation, (i) the closing of the electrical circuit of CE and (ii) the feeding of the used nebulizer with an adequate flow rate. In the beginning of CE-ICP-MS coupling predominantly home-made interface nebulizer constructions were developed and tested for various speciation problems. Now increasingly such laboratory-constructed interfaces are left. Mostly commercial nebulizers are employed being combined with commercially available tee or cross fittings to connect the CE capillary to the electrode, the additional sheath flow, and the nebulizer. Due to the low sample amounts and low flow rates from CE, such nebulizers are typically low-flow nebulizers like, e.g., the microconcentric nebulizer (MCN) and the direct injection nebulizer (DIN). However, there are also reports on couplings using standard Meinhard systems. Still the control and reduction of a siphoning sucting flow and sufficient detection limits are the major problems in hyphenating CE to ICP-MS. Different solutions are reported on these problems and summarized here. Finally numerous applications are reported. Mostly, applications are performed on speciation of selenium, arsenic, metallothionein isoforms, mercury, or cobalt. PMID- 15822065 TI - Determination of glyphosate as cross-contaminant in a commercial herbicide by capillary electrophoresis-electrospray ionization-mass spectrometry. AB - Capillary electrophoresis combined with mass spectrometry (CE-MS) was used for the rapid determination of the negatively charged herbicide, Glyphosate, in a selective dried granule (DG) formulation. The CE-MS method was required to ensure product safety from the risk of cross contamination of a selective herbicide formulation. Glyphosate separation was achieved by using a bare fused-silica capillary column, operated in the reversed-polarity mode, using ammonium formate buffer, pH 2.5. The total CE-MS analysis time was under 10 min and the limit of detection was 10 ng/mL. The CE-MS analysis of Glyphosate was simple, rapid, and selective. The method involved minimal sample handling and was proven to be ideal for cross-contamination investigations in manufacturing samples. PMID- 15822067 TI - Early events of electroporation-mediated intramuscular DNA vaccination potentiate Th1-directed immune responses. AB - BACKGROUND: Application of electrical pulses after DNA injection into muscle increases expression of the encoded genes, and is shown to improve antigen specific immune responses when used for DNA vaccination. In addition, electroporation causes tissue injury and inflammatory reactions. Together with immune stimulatory motifs in the injected DNA these factors may potentiate the immune response by acting as adjuvants for the antigen. Here, we have examined the role of these factors in promoting the efficiency of DNA vaccination. METHODS: We injected a plasmid DNA vector containing the gene Ag85B from M. tuberculosis into mouse quadriceps muscles followed by electroporation. Ag85B was under control of a Tet-responsive promoter, and was expressed either immediately or up to 28 days later by administrating doxycycline to the mice. Delayed expression was combined with injection of non-coding DNA or saline with or without electroporation to examine the ability of these factors to enhance the Ag85B-specific antibody response in the blood and cellular responses in the spleen. Blood samples were analysed with ELISA, while the number of Ag85B specific IFN-gamma- and IL-4-producing spleenocytes was analysed with ELISpot. RESULTS: Delaying Ag85B expression by 5 or 28 days caused lower anti-Ag85B specific IgG2a levels. In contrast, the IgG1 antibody response was not significantly affected. Injection of non-coding DNA followed by electroporation moderately increased the IgG2a response. Delaying the Ag85B expression by 28 days reduced the average number of Ag85B-specific IFN-gamma-producing spleenocytes by over 60%. No significant change in the number of IL-4-producing Ag85B-specific spleenocytes was observed. CONCLUSIONS: These results suggest that DNA and electroporation per se may act as good adjuvants in promoting efficient Th1 directed responses during DNA vaccination. PMID- 15822069 TI - Characterization of the sorption of an anthranilate fungicide in soil using thermal analytical and mineralogical techniques. AB - The sorption of pesticides to soil particles has implications for their distribution and fate in the soil environment. A batch equilibrium technique was used to investigate sorption of the fungicide AEC623892 to intact and hydrogen peroxide-treated whole soils and several particle-size fractions isolated from them. Sorption of AEC623892 to the soil as a whole was low. K(f oc) values measured in the whole soils were 169.2 and 41.9 ml g(-1) for Soil A and Soil B respectively. The highest values of K(f oc) were measured in soil particle-size fractions <53 microm (266.5 ml g(-1) in the 2-20 microm fraction of Soil A; 471.9 ml g(-1) in the 20-53 microm fraction of Soil B). Sorption was most irreversible in the 2-20 microm fractions. Overall, treatment of soil particle-size fractions with hydrogen peroxide resulted in lower values of K(f oc) (112.3 ml g(-1) in Soil A whole soil and 30.9 ml g(-1) in Soil B whole soil). In both soils, the maximum sorption among hydrogen-peroxide-treated samples was observed in the <2 microm fraction (166.6 and 311.0 ml g(-1) for Soil A and Soil B, respectively). Investigation of the mineralogical composition of the soils suggested that the clay mineralogy (dominated by kaolinite and illite) is less likely to account for the differences in sorption observed than differences in the characteristics of the soil organic matter. Thermal analysis of the different soil fractions indicated that hydrogen peroxide treatment preferentially removed aliphatic fractions of organic matter, but had less effect on lignin-like, aromatic fractions. PMID- 15822070 TI - High-resolution multi-parameter DNA flow cytometry enables detection of tumour and stromal cell subpopulations in paraffin-embedded tissues. AB - The accuracy of DNA ploidy measurements of paraffin-embedded tissues is limited by the lack of resolution and the inability to identify the DNA diploid population unequivocally in bimodal DNA histograms. A multi-parameter DNA flow cytometric method has been developed that enables the simultaneous detection of neoplastic and stromal cells in samples from dewaxed 50 microm sections or 2 mm diameter punches of archival tissue blocks. The method combines heat pretreatment in sodium citrate buffer and subsequent enzymatic dissociation with a collagenase/dispase mixture. Cells were simultaneously stained for keratin (FITC), vimentin (R-PE), and DNA (PI) before flow cytometric analysis. The method was applied to 12 paraffin-embedded cervical carcinomas and four colorectal carcinomas. In all cervical cancers, distinct keratin-positive and vimentin positive cell populations were observed. While the exclusive vimentin-positive cell fractions always yielded unimodal DNA content distributions, bimodal distributions were observed for the keratin-positive cell fractions in nine cervical carcinomas, whereas one cervical carcinoma showed three distinct G0G1 populations. Coefficients of variation of the G0G1 peaks ranged from 1.70% to 4.79%. Average background, aggregate, and debris values were 14.7% (vimentin positive fraction) and 33.8% (keratin-positive fraction). Flow sorting confirmed that the exclusively vimentin-positive cell fractions represent different normal stromal and infiltrate cells that can serve as an internal ploidy reference enabling discrimination between DNA hypo-diploid and DNA hyper-diploid tumour cell subpopulations. The neoplastic origin of the keratin-vimentin co-expressing cells from two cervical carcinomas was confirmed by genotyping of flow-sorted samples revealing loss of heterozygosity (LOH) of 6p. This improved method obviates the need for fresh/frozen tumour tissue for high-resolution DNA ploidy measurements and enables the isolation of highly purified tumour subpopulations for subsequent genotyping. PMID- 15822071 TI - Feasibility and safety of the transradial approach for the intracoronary spasm provocation test. AB - An angiography-based spasm provocation test is an accurate diagnostic test of coronary vasospastic angina, but is associated with high patient morbidity, mainly because of the femoral approach and the need for a temporary pacemaker. The purpose of this study was to investigate the safety and feasibility of a transradial ergonovine spasm provocation test. The test was performed prospectively in 174 consecutive patients who were under suspicion of coronary vasospasm at our institution from April 2002 to June 2003. Seventy-eight out of 174 procedures (45%) were performed in an outpatient department. The procedural success rate was 168/174 (96%). All failures were because of access failures, and no major complications were noted. Minor complications were observed in nine patients (severe bradycardia in three, hypotension in two, both in two, and nonsustained ventricular tachycardia in two). The incidence of complications was higher in patients showing prolonged spasm in the right coronary artery. No major local complication was noted other than rebleeding in the puncture site during hemostasis in one patient. The transradial spasm provocation test performed without using a temporary pacemaker may be feasible and safe, with a high success rate and low complication rate as well as low patient morbidity. PMID- 15822072 TI - Field experiments for the evaluation of pesticide spray-drift on arable crops. AB - Two distinct approaches were used to characterise spray-drift during the application of atrazine and alachlor to a maize crop. The first consisted in determining the quantities which did not reach their target. A first experiment was carried in 2001 to improve the sampling method. A second experiment in 2002 showed that losses represented 46 and 38% for atrazine and alachlor, respectively. The second approach was to follow the spatiotemporal evolution of the cloud formed during application. The concentrations observed near the application zone during spraying reached 4.5 microg m(-3) for atrazine and 8.5 microg m(-3) for alachlor. With alachlor these concentrations decreased rapidly when increasing distance from the plot or time following treatment, whereas in the case of atrazine they stabilised rapidly (between 0.5 and 0.3 microg m(-3)) both in space and in time. Deposits around the plot were light and slightly higher for alachlor (from 20 to 130 microg m(-2)). Alachlor was more rapidly diluted in space than atrazine, reflecting a differentiated evolution of physical form during the process. Alachlor, being more volatile than atrazine, is quickly transferred to the gaseous phase which was more rapidly dispersed than aerosols. PMID- 15822073 TI - Bifurcational lesions: searching the solution. PMID- 15822074 TI - Facial characteristics are not distinctive features for the acrofacial dysostosis syndrome type Kennedy-Teebi. PMID- 15822076 TI - Role of surgery in the treatment of motor complications. AB - When medications no longer provide patients with Parkinson's disease a reasonable quality of life due to the presence of levodopa-associated motor fluctuations and dyskinesias, surgical treatment is often pursued. Numerous studies have examined the antiparkinsonian efficacy of procedures currently available, but surprisingly few studies have evaluated their effect on motor response complications in a systematic, controlled manner, using appropriate instruments. Nonetheless, the combined evidence from uncontrolled case series and more recent randomized controlled trials reviewed here indicates that unilateral pallidotomy, bilateral pallidal deep brain stimulation, and bilateral subthalamic deep brain stimulation all substantially alleviate levodopa-induced dyskinesias and, to a lesser extent, motor fluctuations. Incorporation of standardized, validated instruments for the quantification of motor response complications in future surgical study protocols will not only allow more accurate comparison of different interventions but also will help physicians select the most appropriate procedure for their patients. PMID- 15822077 TI - Motivation to change in eating disorder patients: a conceptual clarification on the basis of self-determination theory. AB - OBJECTIVE: The current study critically reviews the different motivational frameworks that are applied in the study of eating disorders and provides a more comprehensive conceptualization of motivation to change on the basis of self determination theory. METHOD: The most important conceptualizations of motivation to change among eating disorder patients are identified. RESULTS: Eating disorder patients' motivation to change has been defined very differently, adding confusion to the field and preventing research from being cumulative. On the basis of self-determination theory we argue (a) that the quality of motivation to change is primarily reflected in the degree of internalization of change rather than by the intrinsic motivation to change; (b) that the internalization of change suggests more than only the change being initiated from within the person (internal motivation) for it requires an acceptance of the personal importance of change; and (c) that, in addition to its quality, the quantity of motivation to change should be considered too. DISCUSSION: These three conceptual issues are applied to the study of motivational dynamics in eating-disordered patients. PMID- 15822078 TI - Night eating syndrome in young adult women: prevalence and correlates. AB - OBJECTIVE: The current study examined the prevalence and clinical significance of night eating syndrome (NES) in a community cohort of Black and White women. METHOD: We assessed 682 Black and 659 White women for NES, eating disorders, and psychiatric symptomatology. RESULTS: The prevalence was 1.6% (22 of 1,341; Blacks [n = 20]; Whites [n = 2]). Comparisons between identified Black women and the remaining Black participants revealed no significant differences in obesity, psychiatric comorbidity, or self-reported psychiatric distress. Comorbidity with eating disorders as outlined in the 4th ed. of the Diagnostic and Statistical Manual of Mental Disorders (Washington, DC: American Psychiatric Association) was low (n = 1 [4.5%]). Black NES women were significantly less likely than Black non NES women to be overweight and significantly more likely to have two or more children. DISCUSSION: NES was rare in this sample of young women. Low comorbidity of NES with other eating disorders suggests that NES may be distinct from the DSM IV recognized eating disorders. Longitudinal data are needed to determine the long-term health implications of this behavioral pattern. PMID- 15822079 TI - Characteristics of anorexia nervosa-related deaths in Norway (1992-2000): data from the National Patient Register and the Causes of Death Register. AB - OBJECTIVE: This study investigated demographic and diagnostic characteristics of individuals whose medical record or death certificate indicated the presence of anorexia nervosa at the time of death. METHOD: Two national registers, the National Patient Register (NPR) and the Causes of Death Register (CODR), were examined in Norway for anorexia nervosa-related deaths occurring across a 9-year period (1992-2000). RESULTS: The medical record or death certificate listed anorexia nervosa as a diagnosis or cause of death for 66 individuals. Rates of death were 6.46 and 9.93 per 100,000 deaths for the NPR and the CODR, respectively. A substantial percentage of deaths (43.9%) in both registers occurred at or above the age of 65 years. For the NPR, the mean age at the time of death was 61 years and 31% of deaths occurred among men. For the CODR, the mean age at the time of death was 49 years and 18% of deaths occurred among men. DISCUSSION: Potential merits and shortcomings of assessing mortality rates using register-based data without linkage to a previously identified clinical sample are discussed. PMID- 15822080 TI - Body dissatisfaction and drive for thinness in young adult twins. AB - OBJECTIVE: We explored correlates of the Eating Disorder Inventory subscales Body Dissatisfaction (BD) and Drive for Thinness (DT) and genetic and environmental influences on these traits. METHOD: In a population-based sample of 4,667 Finnish twins aged 22-27 years, we conducted twin modeling to explore genetic and environmental contributions to body dissatisfaction and drive for thinness. Logistic regression was used for the correlational analysis. RESULTS: Various eating and body size-related factors and psychosomatic symptoms were significantly associated with high body dissatisfaction and drive for thinness in both genders. In women, early puberty onset, early initiation of sexual activity, and multiple sex partners were statistically significant risk factors of body dissatisfaction. In gender-specific univariate twin models, additive genes accounted for 59.4% (95% confidence interval [CI] = 53.2-64.7%) of the variance in body dissatisfaction and for 51.0% (95% CI = 43.7-57.5%) of the variance in drive for thinness among females, but for none of the variance among males. DISCUSSION: There are very distinct gender differences in the heritability patterns of body dissatisfaction and drive for thinness in young adults. PMID- 15822081 TI - The paradox of normal serum albumin in anorexia nervosa: a case report. AB - OBJECTIVE: Anorexia nervosa (AN) is associated with marked decreases in caloric intake and a corresponding reduction in body weight due to abnormal self body image. Although counterintuitive, hypoalbuminemia and vitamin deficiencies are not expected consequences of this disorder. Etiologic considerations for hypoalbuminemia are discussed. METHOD: The case report of a patient with AN and marked hypoalbuminemia is presented and a focused literature review is reported. RESULTS: Hypoalbuminemia was initially attributed to starvation. However, occult infection was ultimately responsible. Serum albumin concentration normalized with antibiotic therapy despite minimal restoration of body weight. DISCUSSION: Hypoalbuminemia should not be considered a characteristic feature of AN even in the setting of progressive weight loss. The presence of other potentially life threatening conditions should be sought, as reduced serum albumin concentration is a marker of inflammation in AN. PMID- 15822083 TI - Anthropomorphic characteristics of centerfold models: trends towards slender figures over time. AB - OBJECTIVE: Anthropomorphic characteristics of centerfold models were analyzed to determine if there has been a trend towards slimmer models over the last 50 years. METHOD: Height and weight, breast, waist, and hip measurements of 559 Playboy centerfold models, obtained from the magazine's website, were analyzed. RESULTS: Regression analyses indicated a gradual and subtle change towards slimness. A latent class analysis yielded five basic types of figures and results indicated that the prevalence of the type of figures being depicted changed over time, with a slightly thinner figure being more common. However, really thin figures were rare until the 2000s and the probability of a model having a body mass index (BMI) < 17 was greater in the 1950s than in the 2000s. DISCUSSION: These findings suggest that there has been a trend towards slimness, but that the changes have been gradual. PMID- 15822084 TI - Anorexia nervosa and obsessive-compulsive disorder in a prepubertal patient with bone dysplasia: a case report. AB - OBJECTIVE: The current article describes the case of a 13-year-old girl with body dysplasia, anorexia nervosa, and obsessive-compulsive disorder (OCD). METHOD: She was given cognitive-behavioral therapy and pharmacologic treatment for the obsessive-compulsive symptomatology and exogenous growth hormone to increase her height. RESULTS: She experienced an adequate weight and height increase and remission of obsessive-compulsive symptomatology, and reestablished adequate social and academic functioning. CONCLUSION: After a follow-up of almost 2 years, she had had her menarche, continued her positive eating habits, and had not relapsed into OCD. PMID- 15822085 TI - Bacterial infections in anorexia nervosa: delayed recognition increases complications. AB - OBJECTIVE: We compared the natural history of bacterial infection in patients with anorexia nervosa (AN) with controls, and assessed which of a range of patient characteristics were associated with infection, fever response, and the rate of infectious complications in AN patients. METHOD: The charts of 311 consecutive hospital admissions of AN patients were reviewed. Patients who had a bacterial infection while in the hospital were compared with the AN patients who did not have an infection, with respect to a range of demographic and disease variables. Fever response and infection complication rate also were evaluated in AN patients with a bacterial infection and in nonanorectic control subjects admitted with a bacterial infection. RESULTS: AN patients with a bacterial infection showed a reduced fever response, were often difficult to diagnose because of fewer signs and symptoms, and infection became more frequent with increasing patient age. DISCUSSION: A reduction in fever response and the signs and symptoms of infection significantly delayed diagnosis in AN patients and increased the complication rate from bacterial infection. We recommend that an increased index of suspicion and an early complete blood count and bacteriologic cultures be adopted for the investigation of bacterial infection in AN patients. PMID- 15822086 TI - Inspiring or dispiriting? The effect of diet commercials on snack food consumption in high school and college-aged women. AB - OBJECTIVE: Recent research offers conflicting views of whether women high in dietary restraint are inspired or demoralized after viewing thin-ideal images. We conducted two studies to explore the impact of diet commercials featuring the thin-ideal image on snack food consumption. METHOD: In Study 1, 91 high school women watched a sad movie interrupted with diet or neutral commercials. In Study 2, 102 college women saw diet commercials early or midway through a sad movie. RESULTS: In Study 1, high restraint participants consumed more snacks after exposure to diet commercials than to neutral commercials. In Study 2, early commercials reduced snacking in low restraint women but not in high restraint women. DISCUSSION: Thin-ideal images embedded in diet commercials do not inspire high restraint women but can modify the eating of low restraint women. PMID- 15822087 TI - Influence of bone density results on adolescents with anorexia nervosa. AB - OBJECTIVE: Not reaching an optimal peak bone mass during adolescence puts young patients with anorexia nervosa (AN) at risk for osteoporosis. Qualitative techniques were employed to determine whether having a bone mineral density (BMD) measurement affected the attitudes and behaviors of young women with AN. METHODS: Nineteen adolescents with AN who had undergone BMD measurements were questioned about the experience of having the test and reactions to the results. Themes were identified and statistical analyses were performed. RESULTS: Participants perceived a normal or low BMD in healthy and unhealthy ways. Although not all healthy feelings led to behavioral change, they appeared to serve as driving forces later in their illness. DISCUSSION: Providing young women with BMD results is not always enough to change unhealthy behaviors and may generate some unhealthy thoughts. However, the experience may lead to positive behavioral changes and result in long-term improvement. Clinicians must be careful and sensitive when presenting these results. PMID- 15822088 TI - Childhood gastrointestinal complaints in women with bulimia nervosa. AB - OBJECTIVE: The current study describes childhood gastrointestinal (GI) complaints in adult women presenting for a treatment trial for bulimia nervosa (BN) and examines the correlates of BN symptomatology. METHOD: The sample comprised 135 women with BN who underwent an assessment including questions regarding childhood GI complaints. Individuals were grouped into one of three categories: no GI complaints, GI complaints with/without constipation (GI complaints), and constipation only. Eating disorder psychopathology was compared across these groups. RESULTS: One third of participants reported GI complaints or constipation only in childhood. Women with GI complaints were younger, and had an earlier onset of BN and self-induced vomiting compared with women with no GI complaints. Trends existed for a younger age of first binge, and a higher frequency of binge eating in this group. DISCUSSION: These findings suggest that individuals with childhood GI complaints and other risk factors for BN may be at greater risk of developing a more severe eating disorder at an earlier age. PMID- 15822089 TI - A comparison of quality of life in obese individuals with and without binge eating disorder. AB - OBJECTIVE: This study investigates whether binge eating disorder (BED) in obese individuals is associated with a greater degree of impairment in quality of life (QOL) than obesity alone. METHOD: Treatment-seeking obese individuals with and without BED were compared on QOL scores using the Impact of Weight on Quality of Life (IWQOL-Lite) questionnaire. RESULTS: With the exception of the Physical Function subscale, obese individuals with BED scored significantly higher than non-BED participants on each of the subscales and on the total scale of the IWQOL Lite. For all participants, body mass index (BMI) was related significantly to scores on the Physical Function and Public Distress subscales of the IWQOL-Lite. DISCUSSION: Obese individuals with BED have impaired functioning on psychosocial aspects of QOL in addition to poorer physical functioning associated with obesity. These findings underscore the pervasive impact of BED in obese individuals, as BED is associated with more impairment than obesity alone. PMID- 15822090 TI - Ethnicity as a protective factor against internalization of a thin ideal and body dissatisfaction. AB - OBJECTIVE: We examined the potential for ethnicity to moderate the relationships between awareness and internalization of sociocultural ideals of appearance and between internalization and body dissatisfaction. METHOD: Spanish (n = 100), Mexican American (n = 100), and European American (n = 100) female participants completed measures of sociocultural attitudes and body dissatisfaction. Path analysis using maximum likelihood with robust standard errors tested the relationships across and within ethnic groups. RESULTS: There was evidence for the mediational effect of internalization on the relationship between awareness and body dissatisfaction. Furthermore, both relationships were significantly stronger for European American women than for Mexican American or Spanish women (the predicted moderator effect). DISCUSSION: Results demonstrate how ethnicity may protect against the development of eating disorder symptoms and suggest that eating disorder prevention should involve denouncing the thin ideal, minimizing appearance as an indicator of value, and emphasizing traits other than appearance as determinants of worth. PMID- 15822092 TI - Eating disorder symptomatology and gender role orientation. AB - OBJECTIVE: The aim of the current study was to investigate associations between gender role orientation and eating pathology in a sample of females with eating disorders (ED). METHODS: Sixty-eight women with anorexia nervosa and 123 women with bulimia nervosa completed the Bem Sex Role Inventory (BSRI), the Eating Disorders Inventory, and the Sexual Anxiety subscale of the Anorexia Nervosa Inventory for Self-Rating. RESULTS: All three subscales of the BSRI--femininity, masculinity, and social desirability--correlated negatively with self-reported ED behavior and attitudes. Individuals with high levels of androgyny (i.e., those scoring high on femininity and masculinity) reported lower levels of ED symptomatology compared with undifferentiated individuals (i.e., those scoring low on femininity and masculinity), who showed higher levels of ED symptoms. DISCUSSION: Femininity and masculinity, although independent traits, should not be considered isolated from each other in the context of ED. In females with ED, androgyny appears to be associated with lower levels of ED symptoms. Self-esteem may play a mediating role in this association. PMID- 15822091 TI - Application of an algorithm-driven protocol to simultaneously provide universal and targeted prevention programs. AB - OBJECTIVE: Our objective was to develop a model to simultaneously prevent eating disorders and weight gain among female high school students. METHOD: Of 188 female 10th graders enrolled in health classes, 174 elected to participate in the current study. They were assessed on-line and decided to participate in one of four interventions appropriate to their risk. RESULTS: The algorithm identified 111 no-risk (NR), 36 eating disorder risk (EDR), 16 overweight risk (OR), and 5 both risks. Fifty-six percent of the EDR and 50% of the OR groups elected to receive the recommended targeted curricula. Significant improvements in weight and shape concerns were observed in all groups. DISCUSSION: An Internet-delivered program can be used to assess risk and provide simultaneous universal and targeted interventions in classroom settings. PMID- 15822093 TI - Serotonin synthesis by two distinct enzymes in Drosophila melanogaster. AB - Annotation of the sequenced Drosophila genome suggested the presence of an additional enzyme with extensive homology to mammalian tryptophan hydroxylase, which we have termed DTRH. In this work, we show that enzymatic analyses of the putative DTRH enzyme expressed in Escherichia coli confirm that it acts as a tryptophan hydroxylase but can also hydroxylate phenylalanine, in vitro. Building upon the knowledge gained from the work in mice and zebrafish, it is possible to hypothesize that DTRH may be primarily neuronal in function and expression, and DTPH, which has been previously shown to have phenylalanine hydroxylation as its primary role, may be the peripheral tryptophan hydroxylase in Drosophila. The experiments presented in this report also show that DTRH is similar to DTPH in that it exhibits differential hydroxylase activity based on substrate. When DTRH uses tryptophan as a substrate, substrate inhibition, catecholamine inhibition, and decreased tryptophan hydroxylase activity in the presence of serotonin synthesis inhibitors are observed. When DTRH uses phenylalanine as a substrate, end product inhibition, increased phenylalanine hydroxylase activity after phosphorylation by cAMP-dependent protein kinase, and a decrease in phenylalanine hydroxylase activity in the presence of the serotonin synthesis inhibitor, alpha methyl-(DL)-tryptophan are observed. These experiments suggest that the presence of distinct tryptophan hydroxylase enzymes may be evolutionarily conserved and serve as an ancient mechanism to appropriately regulate the production of serotonin in its target tissues. PMID- 15822094 TI - Eicosanoids influence in vitro elongation of plasmatocytes from the tobacco hornworm, Manduca sexta. AB - Nodule formation is the predominant insect cellular defense reaction to bacterial challenges, responsible for clearing the largest proportion of infecting bacteria from hemolymph circulation. Hemocyte spreading behavior is a critical step in the nodulation process. It has been suggested that eicosanoids mediate several steps in the process. However, the influence of eicosanoids on hemocyte spreading has not been investigated in detail. To test the hypothesis that eicosanoids mediate hemocyte spreading behavior, I treated larvae of the tobacco hornworm, Manduca sexta, with eicosanoid biosynthesis inhibitors and later assessed plasmatocyte elongation on glass slides. Plasmatocytes from larvae treated with dexamethasone did not elongate to the extent of plasmatocytes from untreated control larvae. The dexamethasone effect on plasmatocyte elongation was expressed in a dose dependent manner and was reversed by injecting dexamethasone-treated larvae with the eicosanoid-precursor fatty acid, arachidonic acid. Palmitic acid, which is not substrate for eicosanoid biosynthesis, did not reverse the influence of dexamethasone on plasmatocyte elongation. Finally, plasmatocytes from larvae treated with a range of eicosanoid biosynthesis inhibitors did not elongate to the extent of plasmatocytes from control larvae. Plasmatocyte width did not appear to be influenced in this study. These findings strongly support the idea that insect plasmatocyte elongation is influenced by eicosanoids. PMID- 15822095 TI - Eicosanoids mediate the laminarin-induced nodulation response in larvae of the flesh fly, Neobellieria bullata. AB - Insects have a highly developed innate immune system, including humoral and cellular components. The cellular immune responses refer to hemocyte-mediated processes such as phagocytosis, nodulation, and encapsulation. Nodulation is considered the predominant defense reaction to infection in insects. Treating third instar larvae of the grey flesh fly, Neobellieria bullata, with laminarin (beta-1,3-glucan, a typical component of fungal cell walls) induced nodulation in a dose-dependent manner. This reaction was initiated very soon after injection and reached its maximal response level after 4 h. The nodules were not randomly distributed in the hemocoel, but were concentrated around the crop. The possible role of eicosanoids in this nodulation process was determined by treating larvae with the phospholipase A(2) inhibitor, dexamethasone, the cyclooxygenase inhibitor, naproxen, and the lipoxygenase inhibitor, esculetin. Both dexamethasone and naproxen significantly impaired the ability of N. bullata larvae to form nodules in response to laminarin. Supplying dexamethasone-treated larvae with the eicosanoid precursor, arachidonic acid, restored the full response. On the other hand, treating larvae with esculetin did not influence the formation of nodules in response to laminarin. This is the first study that demonstrates the occurrence of a laminarin-induced nodulation response in Diptera. Phospholipase A(2) and cyclooxygenase activities, both involved in prostaglandin biosynthesis, appear to play an important role in the regulation of this process. PMID- 15822096 TI - Ligand binding is without effect on complex formation of the ligand binding domain of the ecdysone receptor (EcR). AB - The ligand-binding domain (LBD) encompassing the C-terminal parts of the D- and the complete E-domains of the ecdysteroid receptor (EcR) fused to Gal4(AD) is present in two high molecular weight complexes (600 and 150 kDa) in yeast extracts according to size exclusion chromatography (Superdex 200 HR 10/30). Hormone binding is mainly associated with 150-kDa complexes. Complex formation is not influenced by hormone, but the ligand stabilizes the complexes at elevated salt concentrations. Mutational analysis of Gal4(AD)-EcR(LBD) revealed that formation of 600-kDa, but not 150-kDa, complexes depends on dimerization mediated by the EcR(LBD). Deletion of helix 12 is without effect. Mutation of K497 in helix 4, known to be essential for comodulator binding, abolishes 600-KDa complexes, but does not interfere with the formation of 150-kDa complexes. In contrast, the DE-domains of USP fused to Gal4(DBD) elute as monomer after elimination of the dimerization capacity of the ligand-binding domains by mutation of P463 in helix 10. The data presented here reveal that the complex formation of ligand-binding domains EcR and USP ligand is different. PMID- 15822097 TI - High accuracy prediction of beta-turns and their types using propensities and multiple alignments. AB - We have developed a method that predicts both the presence and the type of beta turns, using a straightforward approach based on propensities and multiple alignments. The propensities were calculated classically, but the way to use them for prediction was completely new: starting from a tetrapeptide sequence on which one wants to evaluate the presence of a beta-turn, the propensity for a given residue is modified by taking into account all the residues present in the multiple alignment at this position. The evaluation of a score is then done by weighting these propensities by the use of Position-specific score matrices generated by PSI-BLAST. The introduction of secondary structure information predicted by PSIPRED or SSPRO2 as well as taking into account the flanking residues around the tetrapeptide improved the accuracy greatly. This latter evaluated on a database of 426 reference proteins (previously used on other studies) by a sevenfold crossvalidation gave very good results with a Matthews Correlation Coefficient (MCC) of 0.42 and an overall prediction accuracy of 74.8%; this places our method among the best ones. A jackknife test was also done, which gave results within the same range. This shows that it is possible to reach neural networks accuracy with considerably less computional cost and complexity. Furthermore, propensities remain excellent descriptors of amino acid tendencies to belong to beta-turns, which can be useful for peptide or protein engineering and design. For beta-turn type prediction, we reached the best accuracy ever published in terms of MCC (except for the irregular type IV) in the range of 0.25-0.30 for types I, II, and I' and 0.13-0.15 for types VIII, II', and IV. To our knowledge, our method is the only one available on the Web that predicts types I' and II'. The accuracy evaluated on two larger databases of 547 and 823 proteins was not improved significantly. All of this was implemented into a Web server called COUDES (French acronym for: Chercher Ou Une Deviation Existe Surement), which is available at the following URL: http://bioserv.rpbs.jussieu.fr/Coudes/index.html within the new bioinformatics platform RPBS. PMID- 15822098 TI - RECOORD: a recalculated coordinate database of 500+ proteins from the PDB using restraints from the BioMagResBank. AB - State-of-the-art methods based on CNS and CYANA were used to recalculate the nuclear magnetic resonance (NMR) solution structures of 500+ proteins for which coordinates and NMR restraints are available from the Protein Data Bank. Curated restraints were obtained from the BioMagResBank FRED database. Although the original NMR structures were determined by various methods, they all were recalculated by CNS and CYANA and refined subsequently by restrained molecular dynamics (CNS) in a hydrated environment. We present an extensive analysis of the results, in terms of various quality indicators generated by PROCHECK and WHAT_CHECK. On average, the quality indicators for packing and Ramachandran appearance moved one standard deviation closer to the mean of the reference database. The structural quality of the recalculated structures is discussed in relation to various parameters, including number of restraints per residue, NOE completeness and positional root mean square deviation (RMSD). Correlations between pairs of these quality indicators were generally low; for example, there is a weak correlation between the number of restraints per residue and the Ramachandran appearance according to WHAT_CHECK (r = 0.31). The set of recalculated coordinates constitutes a unified database of protein structures in which potential user- and software-dependent biases have been kept as small as possible. The database can be used by the structural biology community for further development of calculation protocols, validation tools, structure-based statistical approaches and modeling. The RECOORD database of recalculated structures is publicly available from http://www.ebi.ac.uk/msd/recoord. PMID- 15822099 TI - Clustering algorithms for identifying core atom sets and for assessing the precision of protein structure ensembles. AB - An important open question in the field of NMR-based biomolecular structure determination is how best to characterize the precision of the resulting ensemble of structures. Typically, the RMSD, as minimized in superimposing the ensemble of structures, is the preferred measure of precision. However, the presence of poorly determined atomic coordinates and multiple "RMSD-stable domains"--locally well-defined regions that are not aligned in global superimpositions--complicate RMSD calculations. In this paper, we present a method, based on a novel, structurally defined order parameter, for identifying a set of core atoms to use in determining superimpositions for RMSD calculations. In addition we present a method for deciding whether to partition that core atom set into "RMSD-stable domains" and, if so, how to determine partitioning of the core atom set. We demonstrate our algorithm and its application in calculating statistically sound RMSD values by applying it to a set of NMR-derived structural ensembles, superimposing each RMSD-stable domain (or the entire core atom set, where appropriate) found in each protein structure under consideration. A parameter calculated by our algorithm using a novel, kurtosis-based criterion, the epsilon value, is a measure of precision of the superimposition that complements the RMSD. In addition, we compare our algorithm with previously described algorithms for determining core atom sets. The methods presented in this paper for biomolecular structure superimposition are quite general, and have application in many areas of structural bioinformatics and structural biology. PMID- 15822100 TI - Quantifying allosteric effects in proteins. AB - In allosteric regulation, protein activity is altered when ligand binding causes changes in the protein conformational distribution. Little is known about which aspects of protein design lead to effective allosteric regulation, however. To increase understanding of the relation between protein structure and allosteric effects, we have developed theoretical tools to quantify the influence of protein ligand interactions on probability distributions of reaction rates and protein conformations. We define the rate divergence, Dk, and the allosteric potential, Dx, as the Kullback-Leibler divergence between either the reaction-rate distributions or protein conformational distributions with and without the ligand bound. We then define Dx as the change in the conformational distribution of the combined protein/ligand system, derive Dx in the harmonic approximation, and identify contributions from 3 separate terms: the first term, D[stackxomega], results from changes in the eigenvalue spectrum; the second term, D[stackxDeltax], results from changes in the mean conformation; and the third term, Dxv, corresponds to changes in the eigenvectors. Using normal modes analysis, we have calculated these terms for a natural interaction between lysozyme and the ligand tri-N-acetyl-D-glucosamine, and compared them with calculations for a large number of simulated random interactions. The comparison shows that interactions in the known binding-site are associated with large values of Dxv. The results motivate using allosteric potential calculations to predict functional binding sites on proteins, and suggest the possibility that, in Nature, effective ligand interactions occur at intrinsic control points at which binding induces a relatively large change in the protein conformational distribution. PMID- 15822101 TI - A structural alphabet for local protein structures: improved prediction methods. AB - Three-dimensional protein structures can be described with a library of 3D fragments that define a structural alphabet. We have previously proposed such an alphabet, composed of 16 patterns of five consecutive amino acids, called Protein Blocks (PBs). These PBs have been used to describe protein backbones and to predict local structures from protein sequences. The Q16 prediction rate reaches 40.7% with an optimization procedure. This article examines two aspects of PBs. First, we determine the effect of the enlargement of databanks on their definition. The results show that the geometrical features of the different PBs are preserved (local RMSD value equal to 0.41 A on average) and sequence structure specificities reinforced when databanks are enlarged. Second, we improve the methods for optimizing PB predictions from sequences, revisiting the optimization procedure and exploring different local prediction strategies. Use of a statistical optimization procedure for the sequence-local structure relation improves prediction accuracy by 8% (Q16 = 48.7%). Better recognition of repetitive structures occurs without losing the prediction efficiency of the other local folds. Adding secondary structure prediction improved the accuracy of Q16 by only 1%. An entropy index (Neq), strongly related to the RMSD value of the difference between predicted PBs and true local structures, is proposed to estimate prediction quality. The Neq is linearly correlated with the Q16 prediction rate distributions, computed for a large set of proteins. An "expected" prediction rate QE16 is deduced with a mean error of 5%. PMID- 15822102 TI - Does structural and chemical divergence play a role in precluding undesirable protein interactions? AB - To understand the evolutionary forces establishing, maintaining, breaking, or precluding protein-protein interactions, a comprehensive data set of protein complexes has been analyzed to examine the overlap between protein interfaces and the most conserved or divergent protein surface areas. The most divergent areas tend to be found predominantly away from protein interfaces, although when found at interfaces, they are associated with specific lack of cross-reactivity between close homologues, like in antibody-antigen complexes. Moreover, the amino acid composition of highly variable regions is significantly different from any other protein surfaces. The variable regions present higher structural plasticity as a result of insertions and deletions, and favor charged over hydrophobic residues, a known strategy to minimize aggregation. This suggests that (1) a rapid rate of mutations at these regions might be continuously altering their properties, making difficult the coadaptation, in shape and chemical complementarity, to potential interacting partners; and (2) the existence of some form of selective pressure for variable areas away from interfaces to accumulate charged residues, perhaps as an evolutionary mechanism to increase solubility and minimize undesirable interactions within the crowded cellular environment. Finally, these results are placed into the context of the aberrant oligomerization of sickle cell anemia hemoglobin and prion proteins. PMID- 15822103 TI - O-raffinose crosslinked hemoglobin lacks site-specific chemistry in the central cavity: structural and functional consequences of beta93Cys modification. AB - Reacting human deoxyHbA0 with oxidized raffinose (O-raffinose), a trisaccharide, results in a low oxygen affinity "blood substitute," stabilized in a noncooperative T-conformation and possesses readily oxidizable rhombic heme. In this study, we fractionated the O-raffinose-modified HbA0 heterogeneous polymer (O-R-PolyHbA0) into six distinct fractions with a molecular weight distribution ranging from 64 to approximately 600 kDa using size-exclusion chromatography (SEC). Oxygen equilibrium and kinetics binding parameters of all fractions were nearly identical, reflecting a lack of heterogeneity in ligand binding properties among O-R-PolyHbA0 species (Hill coefficient n equal to 1.0). Several mass spectrometry techniques were used to evaluate undigested and digested HbA0, O-R PolyHbA0, and O-R-PolyHbA0 fractions. Proposed sites of intramolecular crosslinking (i.e., beta1Lys82, beta2Lys82, and beta1Val1) were not found to be the predominant site of crosslinking within the central cavity. Intermolecular crosslinking with O-raffinose results in no discernible site of amino acids modifications with the exception of beta93Cys and alpha104Cys. Based on accessible surface area (ASA) calculations in intact deoxyHbA0, slight conformational changes are required to allow for the S on alpha104Cys to be modified during the reaction with O-raffinose or its partially oxidized product(s). The stabilization of HbA0 in the T-conformation may not be a direct correlate of O-raffinose induced changes, but an indirect consequence of changing hydration in the water-filled central cavity and/or the distal heme pocket leading in the latter case to accelerated iron oxidation. Structural data presented here when taken together with the oxidative instability of O-R-PolyHbA0 may provide some basis for the reported toxicity of this oxygen carrier. PMID- 15822105 TI - Assessing precision and accuracy of protein structures derived from NMR data. PMID- 15822104 TI - Other pharmacological treatments for motor complications and dyskinesias. AB - Controlling motor complications becomes increasingly difficult with disease progression. The "wearing-off" phenomenon is the most-common motor fluctuation. Wearing-off can be treated by dietary manipulation, shortening the dosing interval, substituting sustained-release levodopa, adding amantadine, or monoamine oxidase type B inhibitors, and other options, including catechol-O methyltransferase inhibitors and the approved dopamine agonists addressed in another chapter. The rotigotine constant-delivery system is being developed to treat wearing-off symptoms. Istradefylline (KW-6002), an adenosine A(2A) receptor antagonist, has been studied for wearing-off and the results will be discussed. The on-off fluctuations can be treated with liquid levodopa and the rescue therapy of injectable apomorphine. Patients may also suffer from dyskinesias. Dyskinesias can be treated with small doses of liquefied levodopa-carbidopa, amantadine, and clozapine, an atypical neuroleptic. PMID- 15822106 TI - Nonmotor complications in Parkinson's disease. AB - Parkinson's disease (PD) is most often considered a disorder of movement. Whereas bradykinesia, rigidity, tremor, and postural instability result in disability, nonmotor complications in PD may be of equal or greater significance in some patients. This review will discuss many of the nonmotor complications in PD, including cognitive, autonomic, sleep, and sensory difficulties that may occur. PMID- 15822107 TI - Role of COMT inhibitors and dopamine agonists in the treatment of motor fluctuations. AB - Although levodopa remains the most effective drug for the symptomatic treatment of Parkinson's disease (PD), there are significant limitations to its chronic use. Growing preclinical and clinical evidence suggests that the severity of motor fluctuations is influenced both by PD severity and pulsatile stimulation of striatal dopamine receptors. Current management of motor fluctuations is based primarily on strategies to prolong the effects of dopaminergic stimulation. This prolongation is accomplished either through the use of long-acting dopaminergic drugs or prolonging of the effects of levodopa. During the past decade, the armamentarium of dopamine agonists increased and agents that prolong the plasma half-life of levodopa became available. Furthermore, recent clinical trials provide evidence-based approaches to improve the management of motor fluctuations in patients with advanced and early PD. PMID- 15822108 TI - Pathophysiology of motor fluctuations in Parkinson's disease. AB - Loss of dopamine neurons in Parkinson's disease (PD) initiates a complex stream of effects that results in the development of tremor, bradykinesia, and rigidity. While levodopa remains the most effective drug for the symptomatic treatment of PD, its chronic administration is associated with the development of motor fluctuations and dyskinesias. The risk of developing motor fluctuations has been linked to disease severity, dosage of levodopa, and the age of the patient. A recent body of preclinical data has demonstrated that alterations in dopaminergic tone as well as in treatment patterns results in cellular adaptations, including alterations in gene expression. This body of preclinical data suggests that nonphysiological, pulsatile stimulation of dopamine receptors induces the development of motor fluctuations and dyskinesias and raises the possibility that nonpulsatile stimulation of dopamine receptors (continuous dopaminergic stimulation) might induce fewer fluctuations. We discuss the theory of continuous dopaminergic stimulation and its implications for the management of motor fluctuations in patients with advanced and early PD. PMID- 15822109 TI - Motor fluctuations and dyskinesias in Parkinson's disease: clinical manifestations. AB - Fluctuations in the symptoms of Parkinson's disease (PD), such as wearing-off and on-off effects, and dyskinesias are related to a variety of factors, including duration and dosage of levodopa, age at onset, stress, sleep, food intake, and other pharmacokinetic and pharmacodynamic mechanisms. The majority of patients, particularly those with young onset of PD, experience these levodopa-related adverse effects after a few years of treatment. Assessment of these motor complications is difficult because of the marked clinical variability between and within patients. Daily diaries have been used in clinical trials designed to assess the effects of various pharmacological and surgical interventions on motor fluctuations and dyskinesias. The most common type of dyskinesia, called "peak dose dyskinesia", usually consists of stereotypical choreic or ballistic movements involving the head, trunk, and limbs, and occasionally, the respiratory muscles, whereas tremor and punding are less-common complications. Dystonia is also typically seen in patients with diphasic dyskinesia and wearing-off effect. Recognition of the full spectrum of clinical phenomenology of levodopa-related motor complications is essential for their treatment and prevention. PMID- 15822111 TI - Neuroprotective therapy in Parkinson's disease and motor complications: a search for a pathogenesis-targeted, disease-modifying strategy. AB - The introduction of levodopa in the late 1960s represented a landmark in the therapy of Parkinson's disease (PD). However, motor complications of chronic levodopa therapy have emerged as a major limitation of this otherwise effective therapy. Advancing medical and surgical treatment of these complications has been the main objective of clinical trials during the past few decades. In addition, basic research has focused on better understanding of the mechanisms of motor complications and how to prevent them. Slowing or delaying the progression of the disease delays the need for levodopa therapy; therefore, neuroprotective strategies may play an important role in preventing the onset and reducing the severity of levodopa-related adverse effects. In this introductory review, we present the rationale for current and experimental therapies designed to favorably modify the progression of PD. If implemented early in the course of the disease, such treatments, if found effective, may not only alter the natural progression of the disease but may also delay or minimize motor and nonmotor complications associated with levodopa. PMID- 15822112 TI - How does nitrous oxide reductase interact with its electron donors?--A docking study. AB - Electron transfer reactions are crucial for respiration and denitrification. In this article, we analyze the interaction of nitrous oxide reductase with its electron donors cytochrome c550 and pseudoazurin. Our docking protocol comprises generation of candidate complexes followed by a selection step based on the distance of the donor and acceptor groups in each partner protein. Finally, the structures of the candidate complexes were optimized using a force field calculation, together with a second distance filtering step. The prediction power of this protocol was studied using the crystal structure of the cytochrome c2/photosynthetic reaction center of Rhodobacter sphaeroides as a reference. The results suggest that both cytochrome c550 and pseudoazurin bind at the same hydrophobic surface patch residing near the CuA center of nitrous oxide reductase. The central, well-conserved interaction surface of the donors is hydrophobic, but it is surrounded by numerous lysine side-chains, which interact electrostatically with analogously positioned side-chain carboxylates of the acceptor. The prediction output is an ensemble of energetically similar structures that are rotationally related to each other. While such an ensemble may reflect incomplete prediction power of the docking protocol, it may also manifest a biological situation where there are multiple ways of forming a productive electron transfer complex. Analyses of the predicted structures and the conservation pattern of the amino acid residues suggest the existence of specific electron transfer pathways to and from the CuA center of nitrous oxide reductase. PMID- 15822113 TI - Catheter-based ventricle-coronary vein bypass. AB - The goal of this study was to investigate the feasibility of a catheter-based ventricle-to-coronary vein bypass (VPASS) in order to achieve retrograde myocardial perfusion by a conduit (VSTENT) from the left ventricle (LV) to the anterior interventricular vein (AIV). Percutaneous coronary venous arterialization has been proposed as a potential treatment strategy for otherwise untreatable coronary artery disease. In an acute setting, the VSTENT implant was deployed percutaneously using the VPASS procedure in five swine. Coronary venous flow and pressure patterns were measured before and after VSTENT implant deployment with and without AIV and left anterior descending artery (LAD) occlusion. In a separate chronic pilot study, the VPASS procedure was completed on two animals that had a mid-LAD occlusion or LAD stenosis. At day 30 post-VPASS procedure, left ventriculography and magnetic resonance imaging (MRI) were performed to assess the patency and myocardial viability of the VSTENT implants. Pre-VSTENT implantation, the mid-AIV systolic wedge pressure was significantly lower than LV systolic pressure during AIV blockage (46 +/- 19 vs. 90 +/- 16 mm Hg; P < 0.01). The VSTENT implant deployment was performed without complication and achieved equalization of the AIV and LV systolic pressures and creation of retrograde flow in the distal AIV (maximal flow velocity: 37 +/- 7 cm/sec). At day 30 post-VPASS procedure, left ventriculography showed VSTENT implant patency. MRI perfusion images demonstrated myocardial viability even with an LAD occlusion. Coronary retrograde perfusion using the VPASS procedure is feasible and may represent a potential technique for end-stage myocardial ischemia. PMID- 15822114 TI - Symptomatic local recurrence of prostate carcinoma after radiation therapy. AB - BACKGROUND: Symptomatic local recurrence of prostate carcinoma (SLRPC) after radiation therapy (RT) is associated with morbidity and debilitating symptoms that have a substantial impact on the patient's quality of life. Most reports on the results of RT for localized prostate carcinoma (PC) do not address this endpoint. The objective of this study was to determine the incidence of SLRPC and to identify the risk factors for this endpoint. METHODS: The medical charts of 1006 patients who received RT for localized PC at the University of Texas M. D. Anderson Cancer Center between 1987 and 1997 were reviewed. Local symptoms were defined as hematuria, voiding symptoms, urinary obstruction, and pelvic pain. Progressive symptoms accompanied by either confirmatory histology or cystoscopic findings were attributed to PC. Univariate and multivariate analyses using Cox proportional hazards models were applied to identify risk predictors. RESULTS: Among 964 patients for whom follow-up data were available, 277 patients had prostate-specific antigen (PSA) progression, and 45 patients died of PC during a median follow-up of 9.4 years. In total, 33 patients (3.4%) developed SLRPC. In patients who experienced biochemical progression, the actuarial 5-year incidence of SLRPC was 8.3%. Among the patients who had developed SLRPC, 23 patients (69.7%) died of PC at a median of 25.3 months from the onset of local symptoms. Adverse histologic tumor subtypes (ductal, small cell, and sarcomatoid) were associated significantly with SLRPC (hazard ratio, 8.4; 95% confidence interval, 2.99-23.63). Clinical T classification at diagnosis, Gleason score, and initial PSA level showed a trend toward an increased hazard ratio. CONCLUSIONS: SLRPC after radiotherapy therapy was an uncommon but clinically significant event. Aggressive histologic subtypes were predictive of this endpoint. Clinical T classification, Gleason score, and initial prostate-specific antigen levels also may have predictive value. PMID- 15822115 TI - Pregnancy associated glycoprotein-1, -6, -7, and -17 are major products of bovine binucleate trophoblast giant cells at midpregnancy. AB - Pregnancy associated glycoproteins (PAGs) are extensively glycosylated secretory proteins of ruminant trophoblast cells. In cattle placenta several PAG cDNAs are expressed, but the variety of correspondent proteins and their degree of glycosylation are not well characterized. Thus, we purified PAGs by using a protocol which included a lectin (Vicia villosa agglutinin) affinity chromatography. Due to their specific glycosylation pattern, PAGs derived from binucleate trophoblast giant cells were highly enriched by this protocol. PAGs were purified from cotyledons of 2 day 100 placentas and from a single placenta at day 155 and 180. In all samples three major bands (75; 66; 56 kDa) were detected by one-dimensional SDS-PAGE. Mass-spectrometric analysis identified the 75 kDa band as a mixture of PAG-7 and PAG-6, the 66 kDa band as PAG-1 and the 56 kDa band as PAG-17. N-terminal sequencing of the day 100 sample confirmed the mass spectrometric identifications. Enzymatic release of N-glycans with peptide-N glycanase-F from PAGs reduced the molecular weight to approximately 37 kDa which corresponds to the theoretical molecular mass of PAGs. Limited peptide-N glycanase-F treatment revealed that all four N-glycosylation sites are quantitatively occupied in PAG-1. Compared to PAG-1 the number of potential N glycosylation sites is lower in PAG-17 (three sites) and higher in PAG-6 and -7 (five and six sites, respectively). This suggests that the number of attached N glycans is the main determinant of molecular mass of bovine PAGs. The degree of glycosylation may be a major factor regulating the plasma half life of PAGs. PMID- 15822117 TI - The potential role of gap junction communication between cumulus cells and bovine oocytes during in vitro maturation. AB - Preliminary studies in our laboratory have indicated that modulating cumulus expansion early or late during culture has a profound influence on the subsequent development of cumulus-enclosed oocytes. Our objectives were to evaluate the effect of short term exposure to recombinant human follicle-stimulating hormone (r-hFSH) during in vitro maturation (IVM) on cumulus expansion and developmental competence of bovine oocytes. A highly significant (P < 0.0001) improvement in blastocyst development rate as a proportion of cleaved oocytes after IVM of oocytes was observed in the presence of r-hFSH for the first 6 hr of culture. To demonstrate the importance of the functional coupling between the oocyte and the cumulus compartment during that period of 6 hr, cumulus-oocyte complexes (COCs) were matured with r-hFSH for the first 6 hr followed by 18 hr in presence of 1 heptanol or 1-octanol (gap junction inhibitors) to block the communication between the two. With the coupling inhibitors, the blastocyst yield was significantly decreased (P < 0.05). A brief treatment (30 min) with the weak base methylamine, known to reverse the gap junction inhibitors effect, significantly (P < 0.05) reversed the inhibitory action of these agents on the blastocyst rate. Gap junction communication between the oocyte and surrounding cumulus cells was further studied using microinjection of the fluorescent dye Lucifer Yellow. Morphological evidences (dye transfer) were obtained that support the presence of functional coupling for a longer period with the FSH short exposure. In conclusion, high developmental rates of bovine oocytes can be achieved with a short exposure to r-hFSH. This effect is believed to be mediated through gap junctions as developmental competence of oocytes is compromised by the inhibition of their function. PMID- 15822119 TI - A national agenda for Latino cancer prevention and control. AB - Although cancer is a leading cause of morbidity and premature death among Latinos, there is limited knowledge of cancer-related issues and priorities of greatest significance to the Latino population, the largest minority group in the nation. This information is vital in helping to guide Latino cancer research, training, and awareness efforts at national, regional, and local levels. To help identify cancer issues of greatest relevance to Latinos, Redes En Accion, The National Hispanic/Latino Cancer Network, a major network among the National Cancer Institute's Special Populations Networks, conducted a survey of 624 key opinion leaders from around the country. Respondents were asked to rank the three cancer sites most important to Latinos in their region and the five issues of greatest significance for this population's cancer prevention and control. Recommendations were prioritized for three specific areas: 1) research, 2) training and/or professional education, and 3) awareness and/or public education. Among cancers, breast carcinoma was ranked number one, followed in order by cervical and lung carcinomas. The issues of greatest significance to Latinos were 1) access to cancer screening and care, 2) tobacco use, 3) patient-doctor communication, 4) nutrition, and 5) risk communication. This survey solicited information from scientists, health care professionals, leaders of government agencies, professional and community-based organizations, and other stakeholders in Latino health. The results laid the foundation for a national Redes En Accion Latino cancer agenda, thus providing a useful tool for individuals and organizations engaged in cancer prevention and control efforts among the Hispanic Latino population. PMID- 15822118 TI - Body mass index is weakly associated with, and not a helpful predictor of, disease progression in men with clinically localized prostate carcinoma treated with radical prostatectomy. AB - BACKGROUND: Several studies have recently suggested an association between body mass index (BMI) and disease progression after radical prostatectomy. In the current study, the authors examined this association and that between the reciprocal of BMI (INVBMI, 1/BMI) and progression-free probability in men treated with radical retropubic prostatectomy (RRP) for clinically localized prostate carcinoma. METHODS: The authors retrospectively studied 2210 patients who underwent RRP at Memorial Sloan-Kettering Cancer Center between September 1986 and May 2003. Clinicopathologic variables analyzed included BMI (kg/m2), preoperative serum prostate-specific antigen level (ng/mL), clinical T classification, year of surgery, race, biopsy-derived primary and secondary Gleason grades, and INVBMI, known to better correlate with percent body fat than BMI. Cox regression analysis was used to examine the possible association between BMI or its reciprocal with disease progression after controlling for the effects of common prognostic factors. The areas under the receiver operating curve (AUC) for models with and without INVBMI were calculated. RESULTS: Of the 2210 patients analyzed, 251 experienced disease progression in a median follow-up time of 25.9 months (range, 0-143 months). After adjusting for all clinical variables, both BMI (P = 0.071; hazards ratio [HR] = 1.027) and INVBMI (P = 0.041; HR < 0.001) were associated with disease progression. However, the areas under AUC for models with and without INVBMI were similar (range, 0.794-0.798). CONCLUSIONS: Although conflicting evidence has been reported regarding the link between obesity and an increased risk of developing prostate carcinoma, as well as an increased risk of developing aggressive disease and prostate carcinoma-related mortality, the authors found weak associations with disease progression for both BMI and INVBMI. These variables were of negligible prognostic value in men who received surgery. Studies with longer follow-up, that examine alternative end points, and that follow treatment(s) besides surgery are needed. PMID- 15822120 TI - c-KIT mutation analysis for diagnosis of gastrointestinal stromal tumors in fine needle aspiration specimens. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are a specific clinicopathologic entity characterized by activating mutations in the c-KIT gene. These mutations are associated with increased immunohistochemical staining for the c-KIT protein. Because many GISTs display a definitive response to the KIT inhibitor imatinib, accurate diagnosis of these neoplasms is of great clinical importance. GISTs are the most common mesenchymal neoplasms of the digestive tract and show lineage differentiation along the lines of the interstitial cells of Cajal. The cytomorphology of GISTs has been well described, but the cytologic features are not entirely specific and immunohistochemical staining is frequently used to confirm the diagnosis. However, KIT protein positivity established by immunohistochemistry (IHC) may be unreliable in some instances because of a small number of KIT-negative GISTs and immunohistochemical KIT positivity in some non GIST spindle cell neoplasms of the gastrointestinal tract. Antigen retrieval may be the cause of this spurious positivity. Recently, the demonstration of KIT and PDGFRA mutations has been shown to be more reliable in establishing the diagnosis of GIST than IHC. METHODS: High resolution amplicon melting analysis was performed in a series of eight gastrointestinal stromal tumors sampled by fine needle aspiration (FNA). RESULTS: In five of eight cases, adequate material was obtained for mutational analysis. In four of these, mutations in the c-KIT gene were detected, and in the fifth case a mutation in the PDGFRA gene was detected. CONCLUSIONS: It appears that high resolution amplicon melting analysis can be successfully performed on material obtained by FNA and will show either KIT for PDGFR mutations in the majority of GIST FNA specimens. PMID- 15822121 TI - Does the interval between papanicolaou tests influence the quality of cytology? AB - BACKGROUND: It is commonly believed that the sensitivity of Papanicolaou (Pap) tests decreases with a short interval between cytology samplings. To the authors' knowledge, there is only limited evidence to support this belief. METHODS: For 5055 women in the Atypical Squamous Cells of Undetermined Significance (ASCUS) Low Grade Squamous Intraepithelial Lesion (LSIL) Triage Study (ALTS), the Pap interval was defined as the number of days between the referral Pap smear demonstrating ASCUS or LSIL ("first cytology") and the enrollment liquid-based ("repeat") cytology. The authors investigated the influence of the interval between Pap smears on repeat cytology by examining percentages of abnormal findings, cellularity, and test sensitivity among women diagnosed with histologic grade 3 cervical intraepithelial neoplasia (CIN3) during the 2-year course of the ALTS. In addition, because human papillomavirus (HPV) DNA adjunct testing is now performed, the authors evaluated HPV viral load, which was assayed using residual liquid cytology specimens, in women with CIN3. RESULTS: The Pap interval ranged from 8-30 days in 763 women, 31-60 days in 2317 women, 61-90 days in 1090 women, 91-120 days in 491 women, and 121-184 days in 394 women (mean of 61.3 days; standard deviation of 34 days). Repeat cytologic interpretations of unsatisfactory findings, ASCUS, and high-grade squamous intraepithelial lesion (HSIL) did not appear to vary among the Pap interval groups. However, low-grade cytologic regression occurred with an increasing Pap interval; negative cytology increased from 28.3% (8-30 days) to 41.6% (121-184 days) (P < 0.0001) whereas LSIL cytology decreased (P trend = 0.002). The approximate cellularity of the samples was slightly better in the interval group of 8-30 days (P trend = 0.04). Among women with CIN3, the repeat test sensitivity at a threshold of ASCUS or greater and the HPV DNA viral load was not found to vary by Pap interval (P trend = 0.80 and P trend = 0.36, respectively). CONCLUSIONS: The authors concluded that a short Pap interval (range, 15-120 days) does not significantly affect the quality of liquid-based repeat cytology, nor the viral load tested from a residual liquid-based specimen. PMID- 15822122 TI - Crystal structure of an indigoidine synthase A (IndA)-like protein (TM1464) from Thermotoga maritima at 1.90 A resolution reveals a new fold. PMID- 15822124 TI - Amphipathic alpha-helices in proteins: results from analysis of protein structures. AB - Amphipathic alpha-helices play a crucial role in mediating the interaction of peptides and proteins with membranes. We have analyzed protein structures for the occurrence of 18-residue amphipathic helices. We find several of these alpha helices having average hydrophobic moments and average hydrophobicities that would favor their interaction with membranes. We have analyzed the distribution of net charge, helix length, normalized frequency of occurrence, and propensities of the 20 amino acids in the delineated 18-residue helices. We have observed distinct differences in the frequencies of occurrence of polar and hydrophobic amino acids at positions 1-18 in amphipathic and nonamphipathic helices. There are also differences in propensities of the 20 amino acids to occur at positions 1-18 of amphipathic and nonamphipathic helices. Synthetic peptides corresponding to some of these surface-seeking helices do possess antibacterial and/or hemolytic activities. Knowledge of the distribution of charges in 18-residue surface-seeking amphipathic alpha-helices, as well as propensity of occurrence of amino acids at various positions, would be useful inputs in the de novo design of amphipathic peptides. PMID- 15822123 TI - Age-related performance of human papillomavirus testing used as an adjunct to cytology for cervical carcinoma screening in a population with a low incidence of cervical carcinoma. AB - BACKGROUND: High-risk human papillomavirus (HR-HPV) testing has been proposed as a replacement for cytology or as an adjunct to cytology for primary cervical carcinoma screening. The objective of this study was to assess the age-specific prevalence of HR-HPV infection and the correlation between HR-HPV status and cytologic diagnosis. METHODS: The authors enrolled 7254 women receiving routine cytologic screening in a cross-sectional study that was conducted during 12 months. Cervical samples were collected using liquid-based cytology to perform both Papanicolaou smears and HR-HPV testing. Analyses were performed using age stratification, and the cytologic results were considered as the reference diagnosis for parameter analysis tests. RESULTS: The overall rate of HR-HPV infection was 11.4% (95% confidence interval, 9-12%) and was higher in younger women compared with older women (age < 30 years vs. > or = 30 years; 16% vs. 8.5%, respectively; P < 0.0001). The overall rate of abnormal cytology was 3.2% and, similarly, was more prevalent in younger women (6.1% vs. 2.4%; P < 0.0001). The best balance between sensitivity and specificity for high-grade lesions or worse occurred predominantly in older age groups (age > or = 50 years). CONCLUSIONS: The prevalence of HR-HPV was age-dependent, with the strongest correlation between HR-HPV positivity and disease observed among older women, who potentially may derive the most benefit. PMID- 15822125 TI - Crystal structure of S-adenosylmethionine:tRNA ribosyltransferase-isomerase (QueA) from Thermotoga maritima at 2.0 A resolution reveals a new fold. PMID- 15822126 TI - Clinical report of a pure subtelomeric 1qter deletion in a boy with mental retardation and multiple anomalies adds further evidence for a specific phenotype. AB - Deletions of the 1q telomere have been reported in several studies screening for subtelomeric rearrangements. However, an adequate clinical description is available from only a few patients. We provide a clinical description of a patient with a subtelomeric deletion of chromosome 1q, previously detected by us in a screening study. Comparison of the clinical presentation of our patient with rare cases reported previously provides further evidence for a specific phenotype of 1q patients, including mental retardation, growth retardation, sometimes with prenatal onset, progressive microcephaly, seizures, hand and foot abnormalities and a variety of midline defects, including corpus callosum, cardiac, genital and gastro-esophageal abnormalities. This clinical presentation is reminiscent of that of patients with larger, microscopically visible deletions of chromosome 1q (>3 Mb) characterized by growth and mental retardation, coarse faces with thin upper lip, epilepsy, and variable other anomalies. In addition, the breakpoint region was mapped to a 26 kb region within the RGS7 gene. Among the 17 known genes in the candidate region, are zinc-finger genes. Other members of this gene family have been implicated in different forms of mental retardation. PMID- 15822127 TI - Computational analysis of molecular basis of 1:1 interactions of NRG-1beta wild type and variants with ErbB3 and ErbB4. AB - The neuregulin/ErbB system is a growth factor/receptor cascade that has been proven to be essential in the development of the heart and the sympathetic nervous system. However, the basis of the specificity of ligand-receptor recognition remains to be elucidated. In this study, the structures of NRG 1beta/ErbB3 and NRG-1beta/ErbB4 complexes were modeled based on the available structures of the homologous proteins. The binding free energies of NRG-1beta to ErbB3 and ErbB4 were calculated using the molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) computational method. In addition, computational alanine scanning mutagenesis was performed in the binding site of NRG-1beta and the difference in the binding free energies between NRG-1beta mutants and the receptors was calculated. The results specify the contribution of each residue at the interaction interfaces to the binding affinity of NRG-1beta with ErbB3 and ErbB4, identifying several important interaction residue pairs that are in agreement with previously acquired experimental data. This indicates that the presented structural models of NRG-1beta/ErbB3 and NRG-1beta/ErbB4 complexes are reliable and could be used to guide future studies, such as performing desirable mutations on NRG-1beta to increase the binding affinity and selectivity to the receptor and discovering new therapeutic agents for the treatment of heart failure. PMID- 15822128 TI - Cytologic findings and protein expression profiles associated with ductal carcinoma of the breast in ductal lavage specimens using surface-enhanced laser desorption and ionization-time of flight mass spectrometry. AB - BACKGROUND: Early detection of breast carcinoma enhances the chances for patient survival. The authors' work focused on an innovative technique that couples breast ductal lavage (DL) with surface-enhanced laser desorption and ionization time of flight mass spectrometry (SELDI-TOF MS) to yield a highly sensitive and specific method of breast carcinoma detection. METHODS: The study group included 16 women who had unilateral, biopsy-proven breast carcinoma. Studying paired DL specimens from each woman (the breast with and the breast without carcinoma), a cytologic investigation was performed on the cells present in the DL samples, and the protein content of the DL fluid was analyzed with the SELDI-TOF MS technique using the strong anionic exchange chip surface. RESULTS: Only 5 of 16 DL specimens (31%) from breasts with biopsy-proven carcinoma contained malignant cells, whereas the remaining samples contained only histiocytes and clusters of benign ductal epithelium. In contrast, 12 of 16 DL specimens (75%) from breasts that contained carcinoma had a different protein peak pattern compared with the paired DL specimen from the same patient's contralateral, uninvolved breast. This finding was independent of the presence of neoplastic cells in the lavage fluid. In addition, specific protein peaks, which may represent potential biomarkers, were identified in the DL fluids from breasts with carcinoma. Some of these peaks were conserved between different patients. CONCLUSIONS: The combination of breast DL with SELDI-TOF MS offers a unique and powerful technique for the detection and monitoring of breast carcinoma. This method has the potential to enhance the diagnostic utility of conventional DL cytology. PMID- 15822129 TI - Use of the glycophorin A somatic mutation assay for rapid, unambiguous identification of Fanconi anemia homozygotes regardless of GPA genotype. AB - A 7-year-old girl was hospitalized with pancytopenia requiring blood transfusion. She and an older brother with suspicious symptoms were referred for laboratory testing to confirm a clinical diagnosis of Fanconi anemia (FA). Blood samples from these two children and one parent were examined with the GPA somatic mutation assay. The patient's total GPA somatic mutation frequency of 1.4 x 10( 4) was determined despite the confounding effects of her recent transfusion, and was greater than 10-fold higher than that of a population of pediatric controls, consistent with the known FA phenotype. Her brother was not informative for the standard GPA assay, which requires heterozygosity for the MN blood group, but was analyzed with a modified assay that measured only allele loss mutation. His mutation frequency, 6.8 x 10(-4) was also supportive of a diagnosis of FA. Both analyses also showed evidence of ongoing mutation through terminal erythroblast differentiation, a characteristic of patients with DNA repair syndromes which further confirmed the diagnoses. These conclusions were confirmed with traditional DEB-induced chromosome breakage studies. The quantitative and qualitative aspects of the GPA assay relevant for applying this test for FA diagnosis, and perhaps for carrier detection, are discussed. PMID- 15822130 TI - Three-dimensional conformal radiotherapy for portal vein thrombosis of hepatocellular carcinoma. AB - BACKGROUND: Portal vein thrombosis (PVT) is a common complication in patients with advanced-stage hepatocellular carcinoma (HCC). The authors evaluated the impact of radiotherapy (RT) for PVT of HCC and analyzed the dose-response relation between RT and PVT. METHODS: Between March 1995 and December 2003, 59 patients diagnosed as HCC with PVT were included. The inclusion criteria were unresectable tumor with thrombosis in the main or first branch of the portal vein, liver function of Child-Pugh Class A or B, and an Eastern Cooperative Oncology Group performance status score of 0-2. The median age of the patients was 57 years (range, 36-78 years). A daily dose ranging from 2 to 3 gray (Gy) was administered using 6 or 10-megavolt (MV) X-rays, at 5 fractions a week, to deliver a total dose range of 30-54 Gy, which was a biologic effective dose of 39 70.2 Gy(10) with an alpha/beta ratio of 10. RESULTS: Follow-up computed tomography scans showed a complete response (CR) in 4 of 59 patients (6.8%), a partial response (PR) in 23 patients (39.0%), no response (NR) in 28 patients (47.5%), and progressive disease (PD) in 4 patients (6.8%). The mean RT doses in the responders (CR and PR) and nonresponders (NR and PD) were 59.6 +/- 5.6 Gy(10) and 54.9 +/- 8.5 Gy(10), respectively (P = 0.036). The response rates in patients receiving < 58 Gy(10) and > or = 58 Gy(10) were 20% and 54.6%, respectively (P = 0.034). The median survival duration and the 1-year and 2-year survival rates in the responders were 10.7 months, 40.7%, and 20.7%, respectively, and were 5.3 months, 25.0%, and 4.7%, respectively, in the nonresponders (P = 0.050). CONCLUSIONS: RT induced a 45.8% objective response rate for PVT in patients with HCC. A dose-response relation was found to exist between the RT dose and PVT response. These results suggested that RT may be a treatment option for PVT in patients with HCC and that an RT dose > or = 58 Gy(10) should be recommended. PMID- 15822131 TI - Human papillomavirus genome detection by in situ hybridization in fine-needle aspirates of metastatic lesions from head and neck squamous cell carcinomas. AB - BACKGROUND: Patients with head and neck squamous cell carcinoma (HNSCC) often present with metastatic disease. The diagnosis of metastatic lesions usually is determined by fine-needle aspiration. Human papillomavirus (HPV) is now being considered as a causative agent in a subset of HNSCC. The objectives of this study were, first; to search for the presence of HPV DNA by in situ hybridization (ISH) in metastatic lesions from HNSCC using alcohol-fixed, archival, cytopathologic material; second, to characterize the cytologic features of HPV positive metastatic lesions of HNSCC; and, third, to determine whether there is a correlation between the presence of HPV DNA and the origin of metastatic lesions. METHODS: The authors performed chromogenic ISH analysis for HPV DNA on fine needle aspiration materials from metastatic lesions from 26 patients with HNSCC. Along with the ISH analysis, a detailed cytologic review was performed, and cytopathologic features were recorded. The HPV DNA status in metastatic lesion was correlated with cytopathologic features and primary tumor location. RESULTS: The integration of HPV DNA was visualized microscopically on tumor cell nuclei in 15% of aspirates. The anatomic locations of the study samples were as follows: 16 lymph node aspirates (11 cervical lymph nodes and 5 lymph nodes at other sites other), 5 tracheostomy sites, and 5 miscellaneous sites located on the head and neck area. Cytologic review revealed 13 keratinized and 13 nonkeratinized metastatic tumors. HPV DNA was detected in four metastatic sites (three lymph nodes and one tracheostomy site). All HPV DNA-positive tumors were of the nonkeratinizing type (P < 0.05; Fisher exact test). The origins of HPV-positive tumors included two laryngeal sites, one nasopharyngeal site, and one oral cavity site. CONCLUSIONS: The current findings showed that archival cytology slides can be used for HPV DNA detection with ISH. The results also showed that HPV DNA containing HNSCC has distinctively nonkeratinizing cytologic features. The authors concluded that HPV DNA not only is involved in the initiation of tumoral processes but also plays an important role in the development of metastatic disease. PMID- 15822133 TI - Ternary nets formed by self-assembly of triangles, squares, and tetrahedra. PMID- 15822132 TI - Body composition and time course changes in regional distribution of fat and lean tissue in unselected cancer patients on palliative care--correlations with food intake, metabolism, exercise capacity, and hormones. AB - BACKGROUND: Several investigations that yielded different results in terms of net changes in body composition of weight-losing cancer patients have been reported that employed a variety of methods based on fundamentally different technology. Most of those reports were cross-sectional, whereas to the authors' knowledge there is sparse information available on longitudinal follow-up measurements in relation to other independent methods for the assessment of metabolism and performance. METHODS: For the current report, the authors evaluated time course changes in body composition (dual-energy X-ray absorptiometry) with measurements of whole body and regional distribution of fat and lean tissue in relation to food and dietary intake, host metabolism (indirect calorimetry), maximum exercise capacity (walking test), and circulating hormones in cancer patients who were receiving palliative care during 4-62 months of follow-up. The entire cohort comprised 311 patients, ages 68 years +/- 3 years who were diagnosed with solid gastrointestinal tumors (84 colorectal tumors, 74 pancreatic tumors, 73 upper gastrointestinal tumors, 51 liver-biliary tumors, 3 breast tumors, 5 melanomas, and 21 other tumor types). RESULTS: Decreased body weight was explained by loss of body fat, preferentially from the trunk, followed by leg tissue and arm tissue, respectively. Lean tissue (fat-free mass) was lost from arm tissue, whereas trunk and leg tissue compartments increased, all concomitant with declines in serum albumin, increased systemic inflammation (C-reactive protein, erythrocyte sedimentation rate), increased serum insulin, and elevated daily caloric intake; whereas serum insulin-like growth factor 1 (IGF-1), resting energy expenditure, and maximum exercise capacity remained unchanged in the same patients. Serum albumin levels (P < 0.001), whole body fat (P < 0.02), and caloric intake (P < 0.001) predicted survival, whereas lean tissue mass did not. Daily intake of fat and carbohydrate was more important for predicting survival than protein intake. Survival also was predicted by serum IGF-1, insulin, leptin, and ghrelin levels (P < 0.02 - P < 0.001). Serum insulin, leptin, and ghrelin (total) levels predicted body fat (P < 0.001), whereas IGF-1 and thyroid hormone levels (T3, free T3) predicted lean tissue mass (P < 0.01). Systemic inflammation primarily explained variation in lean tissue and secondarily explained loss in body fat. Depletion of lean arm tissue was related most to short survival compared with the depletion of lean leg and trunk tissue. CONCLUSIONS: The current results demonstrated that body fat was lost more rapidly than lean tissue in progressive cancer cachexia, a phenomenon that was related highly to alterations in the levels of circulating classic hormones and food intake, including both caloric amount and diet composition. The results showed importance in the planning of efficient palliative treatment for cancer patients. PMID- 15822134 TI - Photoinduced processes within compact dyads based on triphenylpyridinium functionalized bipyridyl complexes of ruthenium(II). AB - As an alternative to conventional charge-separation functional molecular models based on long-range ET within redox cascades, a "compact approach" has been examined. To this end, spacer elements usually inserted between main redox-active units within polyad systems have been removed, allowing extended rigidity but at the expense of enhanced intercomponent electronic communication. The molecular assemblies investigated here are of the P-(theta (1))-A type, where the theta (1) twist angle is related to the degree of conjugation between the photosensitizer (P, of {Ru(bpy)(3)}(2+) type) and the electron-acceptor (A). 4-N- and 4-N-,4'-N (2,4,6-triphenylpyridinio)-2,2'-bipyridine ligands (A(1)-bpy and A(2)-bpy, respectively) have been synthesized to give complexes with Ru(II), 1-bpy and 2 bpy, respectively. Combined solid-state analysis (X-ray crystallography), solution studies ((1)H NMR, cyclic voltammetry) and computational structural optimization allowed verifying that theta (1) angle approaches 90 degrees within 1-bpy and 2-bpy in solution. Also, anticipated existence of strong intercomponent electronic coupling has been confirmed by investigating electronic absorption properties and electrochemical behavior of the compounds. The capability of 1-bpy and 2-bpy to undergo PET process was evaluated by carrying out their photophysical study (steady state emission and time-resolved spectroscopy at both 293 and 77 K). The conformational dependence of photoinduced processes within P (theta (1))-A systems has been established by comparing the photophysical properties of 1-bpy (and 2-bpy) with those of an affiliated species reported in the literature, 1-phen. A complementary theoretical analysis (DFT) of the change of spin density distribution within model [1-bpy(theta (1))](-) mono-reduced species as a function of theta (1) has been undertaken and the possibility of conformationally switching emission properties of P was derived. PMID- 15822135 TI - Structure and unique interactions with DNA of a cationic trans-platinum complex with the nonplanar bicyclic piperidinopiperidine ligand. PMID- 15822136 TI - An old target revisited: two new privileged skeletons and an unexpected binding mode for HIV-protease inhibitors. PMID- 15822137 TI - Proton sandwiches: nonclassical carbocations with tetracoordinate protons. PMID- 15822138 TI - Hydroamination/hydrosilylation sequence catalyzed by titanium complexes. PMID- 15822139 TI - Simultaneous removal of nitrate and heavy metals by iron metal. AB - Great attention should be paid now to simultaneously removing common pollutants, especially inorganic pollutants such as nitrate and heavy metals, as individual removal has been investigated extensively. Removing common pollutants simultaneously by iron metal is a very effective alternative method. Near neutral pH, heavy metals, such as copper and nickel, can be removed rapidly by iron metal, while nitrate removal very much slower than that of copper and nickel, and copper can accelerate nitrate removal when both are removed simultaneously. Even a little amount of copper can enhance nitrate removal efficiently. Different mechanisms of these contaminants removal by iron metal were also discussed. PMID- 15822140 TI - Accumulation and ultrastructural distribution of copper in Elsholtzia splendens. AB - Copper accumulation and intracellular distribution in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species, was investigated by transmission electron microscope (TEM) and gradient centrifugation techniques. Copper concentrations in roots, stems and leaves of E. splendens increased with increasing Cu levels in solution. After exposure to 500 micromol/L Cu for 8 d, about 1000 mg/kg Cu were accumulated in the stem and 250 mg/kg Cu in the leaf of E. splendens. At 50 micromol/L Cu, no significant toxicity was observed in the chloroplast and mitochondrion within its leaf cells, but separation appeared at the cytoplasm and the cell wall within the root cells. At >250 micromol/L Cu, both root and leaf cell organelles in E. splendens were damaged heavily by excessive Cu in vivo. Copper subcellular localization in the plant leaf after 8 days' exposure to 500 micromol/L Cu using gradient centrifugation techniques was found to be decreased in the order: chloroplast>cell wall>soluble fraction>other organelles. The plant root cell wall was found to be the site of highest Cu localization. Increase of Cu exposure time from 8 d to 16 d, increased slightly Cu concentration in cell wall fraction in roots and leaves, while that in the chloroplast fraction decreased in leaves of the plants grown in both 0.25 micromol/L and 500 micromol/L Cu. TEM confirmed that much more Cu localized in cell walls of E. splendens roots and leaves, but also more Cu localized in E. splendens' chloroplast when the plant is exposed to Cu levels>250 micromol/L, as compared to those in the plant grown in 0.25 micromol/L Cu. Copper treatment at levels>250 micromol/L caused pronounced damage in the leaf chloroplast and root organelles. Copper localization in cell walls and chloroplasts could mainly account for the high detoxification of Cu in E. splendens. PMID- 15822141 TI - Ozonation with ultrasonic enhancement of p-nitrophenol wastewater. AB - Synergetic effects for p-nitrophenol degradation were observed in the ozonation with ultrasonic enhancement. The enhancements of removal rate for p-nitrophenol and TOC were around 116% and 294% respectively in comparison with the individual ultrasound and ozonation systems. The synergetic phenomenon is attributed to two physicochemical mechanisms: (1) Ultrasound decomposes ozone causing augmentation of the activity of free radicals; (2) Ultrasonic wave increased the concentration of O(3) in solution because of ultrasonic dispersion. PMID- 15822142 TI - Toxicity of cadmium to soil microbial biomass and its activity: effect of incubation time on Cd ecological dose in a paddy soil. AB - Cadmium (Cd) is ubiquitous in the human environment and has toxic effect on soil microbial biomass or its activity, including microbial biomass carbon (C(mic)), dehydrogenase activity (DHA) and basal respiration (BR), etc., C(mic), DHA, BR were used as bioindicators of the toxic effect of Cd in soil. This study was conducted to determine the effects of Cd on soil microbial biomass and its activity in a paddy soil. The inhibition of microbial biomass and its activity by different Cd concentrations was described by the kinetic model (M1) and the sigmoid dose-response model (M2) in order to calculate three ecological doses of Cd: ED(50), ED(10) and ED(5). Results showed that M2 was better fit than M1 for describing the ecological toxicity dose effect of cadmium on soil microbial biomass and its activity in a paddy soil. M2 for ED values (mg/kg soil) of C(mic), DHA, BR best fitted the measured paddy soil bioindicators. M2 showed that all ED values (mg/kg) increased in turn with increased incubation time. ED(50), ED(10) and ED(5) of C(mic) with M2 were increased in turn from 403.2, 141.1, 100.4 to 1000.7, 230.9, 144.8, respectively, after 10 d to 60 d of incubation. ED(50), ED(10) and ED(5) of DHA with M2 increased in turn from 67.6, 6.2, 1.5 to 101.1, 50.9, 41.0, respectively, after 10 d to 60 d of incubation. ED(50), ED(10) and ED(5) of BR with M2 increased in turn from 149.7, 6.5, 1.8 to 156.5, 50.8, 35.5, respectively, after 10 d to 60 d of incubation. So the ecological dose increased in turn with increased incubation time for M2 showed that toxicity of cadmium to soil microbial biomass and its activity was decreased with increased incubation time. PMID- 15822143 TI - Water and heat transport in hilly red soil of southern China: I. Experiment and analysis. AB - Studies on coupled transfer of soil moisture and heat have been widely carried out for decades. However, little work has been done on red soils, widespread in southern China. The simultaneous transfer of soil moisture and heat depended on soil physical properties and the climate conditions. Red soil is heavy clay and high content of free iron and aluminum oxide. The climate conditions are characterized by the clear four seasons and the serious seasonal drought. The great air temperature differences annually and diurnally result in significant fluctuation in soil temperature in top layer. The closed and evaporating columns experiments with red soil were conducted to simulate the coupled transfer of soil water and heat under the overlaying and opening fields' conditions, and to analyze the effects of soil temperature gradient on the water transfer and the effects of initial soil water contents on the transfer of soil water and heat. The closed and evaporating columns were designed similarly with about 18 degrees C temperatures differences between the top and bottom boundary, except of the upper end closed or exposed to the air, respectively. Results showed that in the closed column, water moved towards the cold end driven by temperature gradient, while the transported water decreased with the increasing initial soil water content until the initial soil water content reached to field capacity equivalent, when almost no changes for the soil moisture profile. In the evaporating column, the net transport of soil water was simultaneously driven by evaporation and temperature gradients, and the drier soil was more influenced by temperature gradient than by evaporation. In drier soil, it took a longer time for the temperature to reach equilibrium, because of more net amount of transported water. PMID- 15822144 TI - Water and heat transport in hilly red soil of southern China: II. Modeling and simulation. AB - Simulation models of heat and water transport have not been rigorously tested for the red soils of southern China. Based on the theory of nonisothermal water-heat coupled transfer, a simulation model, programmed in Visual Basic 6.0, was developed to predict the coupled transfer of water and heat in hilly red soil. A series of soil column experiments for soil water and heat transfer, including soil columns with closed and evaporating top end, were used to test the simulation model. Results showed that in the closed columns, the temporal and spatial distribution of moisture and heat could be very well predicted by the model, while in the evaporating columns, the simulated soil water contents were somewhat different from the observed ones. In the heat flow equation from Taylor and Lary (1964), the effects of soil water evaporation on the heat flow is not involved, which may be the main reason for the differences between simulated and observed results. The predicted temperatures were not in agreement with the observed one with thermal conductivities calculated by de Vries and Wierenga equations, so that it is suggested that K(h), soil heat conductivity, be multiplied by 8.0 for the first 6.5 h and by 1.2 later on. Sensitivity analysis of soil water and heat coefficients showed that the saturated hydraulic conductivity, K(S), and the water diffusivity, D(theta), had great effects on soil water transport; the variation of soil porosity led to the difference of soil thermal properties, and accordingly changed temperature redistribution, which would affect water redistribution. PMID- 15822145 TI - Multi-stage ponds-wetlands ecosystem for effective wastewater treatment. AB - The performance of the Dongying multi-stage ponds-wetlands ecosystem was investigated in this work. Study of the removal of different pollutants (BOD(5), COD, SS, TP, TN, NH(3)-N, etc.) in different temperature seasons and different units in this system indicated that effluent BOD(5) and SS were constant to less than 11 mg/L and 14 mg/L throughout the experimental processes; but that the removal efficiencies of pollutants such as TP, TN, NH(3)-N, COD varied greatly with season. The higher the temperature was, the higher was the observed removal in this system. Additionally, each unit of the system functioned differently in removing pollutants. BOD(5) and SS were mainly removed in the first three units (hybrid facultative ponds, aeration ponds and aerated fish ponds), whereas nitrogen and phosphates were mainly removed in hydrophyte ponds and constructed reed wetlands. The multi-stage ponds-wetlands ecosystem exhibits good potential of removing different pollutants, and the effluent quality meet several standards for wastewater reuse. PMID- 15822146 TI - Adsorption of ruthenium ions on activated charcoal: influence of temperature on the kinetics of the adsorption process. AB - Influence of temperature on ruthenium adsorption on activated charcoal from 3 mol/L HNO(3) solutions was investigated in the temperature range of 288 K to 308 K. It was observed that the rise in temperature increases the adsorption of ruthenium ions on activated charcoal and follows the kinetics of first order rate law with rate constant values 0.0564-0.0640 min(-1) in the temperature range of 288 K to 308 K respectively. The activation energy for the adsorption process was found to be 1.3806 kJ/mol. Various thermodynamics quantities namely H, S and G were computed from the equilibrium constant K(C) values. The results indicated a positive heat of adsorption, a positive S and a negative G. PMID- 15822147 TI - Experimental investigation of external explosion in the venting process. AB - Experimental investigations were conducted on the process of combustion and explosion vent in a 200 mm (diameter) x 400 mm (length) vertical cylindrical vessel. When CH(4)-air mixture gases were used and the vent diameter was 55 mm, conditions of Phi (equivalent ratio)=0.8, Phi=1.0 and Phi=1.3 and two ignition positions (at the cylinder center and bottom) were selected. The venting processes and the correlated factors are discussed in this paper. PMID- 15822148 TI - Influence of surfactant concentration on counter-ion induced solubility of poly(pyridine-2,5-diyl). AB - Protonating the pyridine rings of poly(pyridine-2,5-diyl) with dodecybenzenesulfonic acid and camphorsulphonic acid produces polymer materials which can be dissolved in chloroform (in contrast to the unprotonated polymer, which can only be dissolved in strong acids such as formic acid) and allows mixing the protonated polymers with other chloroform soluble conjugated polymers for use in electronic devices. The protonating behavior of poly(pyridine-2,5 diyl) with two kinds of surfactants is different in some levels. Dodecybenzenesulfonic acid has higher protonating ability than camphorsulphonic acid. PMID- 15822149 TI - Growth mechanism of a preferentially oriented mordenite membrane. AB - A preferentially oriented mordenite membrane was successfully prepared on a seeded porous a-alumina support. Characterization results of XRD (X-ray diffractometer) and FE-SEM (field emission scanning electron microscope) revealed that evolutionary selection might predominantly contribute to the formation of the sharply oriented mordenite membrane. The necessary conditions under which evolutionary selection occurs are: (a) the number density of nuclei on the support surface should be high enough at the early stage; (b) the crystals should grow fastest along one direction; and (c) the zeolite layer should proceed via the successive growth of the crystals nucleated on the support surface instead of the accumulation of the crystals formed in the bulk solution. PMID- 15822150 TI - Synthesis of acetals and ketals catalyzed by tungstosilicic acid supported on active carbon. AB - Catalytic activity of activated carbon supported tungstosilicic acid in synthesizing 2-methyl-2-ethoxycarbonylmethyl- 1,3-dioxolane, 2,4-dimethyl-2 ethoxycarbonylmethyl-1,3-dioxolane, cyclohexanone ethylene ketal, cyclohexanone 1,2-propa- nediol ketal, butanone ethylene ketal, butanone 1,2-propanediol ketal, 2-phenyl-1,3-dioxolane, 4-methyl-2-phenyl-1,3-dioxolane, 2-propyl-1,3-dioxolane, 4-methyl-2-propyl-1,3-dioxolane was reported. It has been demonstrated that activated carbon supported tungstosilicic acid is an excellent catalyst. Various factors involved in these reactions were investigated. The optimum conditions found were: molar ratio of aldehyde/ketone to glycol is 1/1.5, mass ratio of the catalyst used to the reactants is 1.0%, and reaction time is 1.0 h. Under these conditions, the yield of 2-methyl-2-ethoxycarbonylmethyl-1,3-dioxolane is 61.5%, of 2,4-dimethyl- 2-ethoxycarbonylmethyl-1,3-dioxolane is 69.1%, of cyclohexanone ethylene ketal is 74.6%, of cyclohexanone 1,2-propanediol ketal is 80.1%, of butanone ethylene ketal is 69.5%, of butanone 1,2-propanediol ketal is 78.5%, of 2-phenyl-1,3-dioxolane is 56.7%, of 4-methyl-2-phenyl-1,3-dioxolane is 86.2%, of 2-propyl-1,3-dioxolane is 87.5%, of 4-methyl-2-propyl-1,3-dioxolane is 87.9%. PMID- 15822151 TI - Hydrogenation of ortho-nitrochlorobenzene on activated carbon supported platinum catalysts. AB - Platinum/carbon catalyst is one of the most important catalysts in hydrogenation of ortho-nitrochlorobenzene to 2,2'-dichlorohydrazobenzene. The preparation process and the supports of catalysts are studied in this paper. Raw materials and preparation procedure of the activated carbon have great influences on the compositions and surface structure of platinum/carbon catalysts. Platinum catalysts supported on activated carbon with high purity, high surface area, large pore volume and appropriate pore structure usually exhibit higher activities for hydrogenation of ortho-nitrochlorobenzene to 2,2' dichlorohydrazobenzene. The catalyst prepared from H(2)PtCl(6) with pH=3 shows greater catalytic performance than those prepared under other conditions. PMID- 15822153 TI - Effect of seedling age and water depth on morphological and physiological aspects of transplanted rice under high temperature. AB - To study the effect of high temperature, rice seedlings 20, 30, 40 and 50 d were kept at 5, 10, 15 and 20 cm water depth in a water pool. Meteorological findings indicated that water temperature varied up to 10 cm but became stable below this depth. Deep water inflicted higher tiller mortality, minimal increase in dry weight of aerial parts and leaf area, decrease in root length, and decrease in root dry weight especially at 20 cm water depth and produced an unbalanced T/R ratio (top versus root dry weight). However, deep water tended to increase plant length. These parameters, however, excel in shallow water. Older seedlings, with the exception of root dry weight, could not perform well compared to young seedlings in all physiological and morphological aspects. The study revealed that seedlings, particularly young ones, stand well in shallow water and can cope with high temperature. PMID- 15822152 TI - Identification of candidate genes for drought stress tolerance in rice by the integration of a genetic (QTL) map with the rice genome physical map. AB - Genetic improvement for drought stress tolerance in rice involves the quantitative nature of the trait, which reflects the additive effects of several genetic loci throughout the genome. Yield components and related traits under stressed and well-water conditions were assayed in mapping populations derived from crosses of AzucenaxIR64 and AzucenaxBala. To find the candidate rice genes underlying Quantitative Trait Loci (QTL) in these populations, we conducted in silico analysis of a candidate region flanked by the genetic markers RM212 and RM319 on chromosome 1, proximal to the semi-dwarf (sd1) locus. A total of 175 annotated genes were identified from this region. These included 48 genes annotated by functional homology to known genes, 23 pseudogenes, 24 ab initio predicted genes supported by an alignment match to an EST (Expressed sequence tag) of unknown function, and 80 hypothetical genes predicted solely by ab initio means. Among these, 16 candidate genes could potentially be involved in drought stress response. PMID- 15822154 TI - Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy. AB - Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system. PMID- 15822155 TI - A hybrid neural network system for prediction and recognition of promoter regions in human genome. AB - This paper proposes a high specificity and sensitivity algorithm called PromPredictor for recognizing promoter regions in the human genome. PromPredictor extracts compositional features and CpG islands information from genomic sequence, feeding these features as input for a hybrid neural network system (HNN) and then applies the HNN for prediction. It combines a novel promoter recognition model, coding theory, feature selection and dimensionality reduction with machine learning algorithm. Evaluation on Human chromosome 22 was approximately 66% in sensitivity and approximately 48% in specificity. Comparison with two other systems revealed that our method had superior sensitivity and specificity in predicting promoter regions. PromPredictor is written in MATLAB and requires Matlab to run. PromPredictor is freely available at http://www.whtelecom.com/Prompredictor.htm. PMID- 15822156 TI - Statistical properties of nucleotide clusters in DNA sequences. AB - Using the complete genome of Plasmodium falciparum 3D7 which has 14 chromosomes as an example, we have examined the distribution functions for the amount of C or G and A or T consecutively and non-overlapping blocks of m bases in this system. The function P(S) about the number of the consecutive C-G or A-T content cluster conforms to the relation P(S) proportional, variante(-alphas); values of the scaling exponent alpha(CG) are much larger than alpha(AT); and alpha(AT) of 14 chromosomes are hardly changed, whereas alpha(CG) of 14 chromosomes have a number of fluctuations. We found maximum value of A-T cluster size is much larger than C G, which implies the existence of large A-T cluster. Our study of the width function xi(m) of cluster C-G content showed that follows good power law xi(m) proportional, variantm(-gamma). The average gamma for 14 chromosomes is 0.931. These investigations provide some insight into the nucleotide clusters of DNA sequences, and help us understand other properties of DNA sequences. PMID- 15822157 TI - A genetic diversity comparison between captive individuals and wild individuals of Elliot's Pheasant (Syrmaticus ellioti) using mitochondrial DNA. AB - Maintaining genetic diversity is a major issue in conservation biology. In this study, we demonstrate the differences of genetic diversity levels between wild and captive individuals of Elliot's Pheasant Syrmaticus ellioti. Wild individuals showed a higher genetic diversity level than that of the captive individuals. Nucleotide diversity and haplotype diversity of wild individuals were 0.00628 and 0.993, while those of captive individuals were 0.00150 and 0.584 respectively. Only 3 haplotypes of mtDNA control region sequence were identified among 36 captive individuals, while 16 unique haplotypes were identified among the 17 wild individuals in this study. One captive haplotype was shared by a wild individual from Anhui Province. It is concluded that a low number of founders was the likely reason for the lower level genetic diversity of the captive group. Careful genetic management is suggested for captive populations, particularly of such an endangered species, to maintain genetic variability levels. PMID- 15822158 TI - Modeling water and carbon fluxes above summer maize field in North China Plain with back-propagation neural networks. AB - In this work, datasets of water and carbon fluxes measured with eddy covariance technique above a summer maize field in the North China Plain were simulated with artificial neural networks (ANNs) to explore the fluxes responses to local environmental variables. The results showed that photosynthetically active radiation (PAR), vapor pressure deficit (VPD), air temperature (T) and leaf area index (LAI) were primary factors regulating both water vapor and carbon dioxide fluxes. Three-layer back-propagation neural networks (BP) could be applied to model fluxes exchange between cropland surface and atmosphere without using detailed physiological information or specific parameters of the plant. PMID- 15822159 TI - Biomass and microbial activity in a biofilter during backwashing. AB - Biomass and microbial activity in backwashing processes of a biofilter for tertiary treatment were investigated. The microbial groups revealed new distribution along the biofilter depth after low flow rate backwashing for a short time. Then the start-up process was accelerated by backwashing. The biomass profile and microbial activity profile both varying with depth before and after backwashing, can be mathematically described by quadratic equations. Using the profiles, the difference of oxygen demand can be calculated to determine the airflow rate during backwashing. Combined with the difference between biofilters and rapid gravity filters, analysis of biomass and microbial activity can determine more accurately the required airflow rate during backwashing. PMID- 15822160 TI - Restaurant emissions removal by a biofilter with immobilized bacteria. AB - Pseudomonas sp. ZD8 isolated from contaminated soil was immobilized with platane wood chips to produce packing materials for a novel biofilter system utilized to control restaurant emissions. The effects of operational parameters including retention time, temperature, and inlet gas concentration on the removal efficiency and elimination capacity were evaluated. Criteria necessary for a scale-up design of the biofilter was established. High and satisfactory level of rapeseed oil smoke removal efficiency was maintained during operation and the optimal retention time was found to be 18 s corresponding to smoke removal efficiency greater than 97%. The optimal inlet rapeseed oil smoke loading was 120 mg/(m(3) x h) at the upper end of the linear correlation between inlet loading and elimination capacity. PMID- 15822161 TI - Use of 3-D magnetic resonance electrical impedance tomography in detecting human cerebral stroke: a simulation study. AB - We have developed a new three dimensional (3-D) conductivity imaging approach and have used it to detect human brain conductivity changes corresponding to acute cerebral stroke. The proposed Magnetic Resonance Electrical Impedance Tomography (MREIT) approach is based on the J-Substitution algorithm and is expanded to imaging 3-D subject conductivity distribution changes. Computer simulation studies have been conducted to evaluate the present MREIT imaging approach. Simulations of both types of cerebral stroke, hemorrhagic stroke and ischemic stroke, were performed on a four-sphere head model. Simulation results showed that the correlation coefficient (CC) and relative error (RE) between target and estimated conductivity distributions were 0.9245+/-0.0068 and 8.9997%+/-0.0084%, for hemorrhagic stroke, and 0.6748+/-0.0197 and 8.8986%+/-0.0089%, for ischemic stroke, when the SNR (signal-to-noise radio) of added GWN (Gaussian White Noise) was 40. The convergence characteristic was also evaluated according to the changes of CC and RE with different iteration numbers. The CC increases and RE decreases monotonously with the increasing number of iterations. The present simulation results show the feasibility of the proposed 3-D MREIT approach in hemorrhagic and ischemic stroke detection and suggest that the method may become a useful alternative in clinical diagnosis of acute cerebral stroke in humans. PMID- 15822163 TI - PDAs okay for pacemakers, but zip the zapper. PMID- 15822164 TI - Please don't let this be the wave of the future. PMID- 15822162 TI - Pharmacokinetics of gemcitabine in Chinese patients with non-small-cell lung cancer. AB - To determine the pharmacokinetics of gemcitabine (2',2'-difluorodeoxycytidine) in Chinese non-small-cell lung cancer (NSCLC) patients. Six study subjects were administered gemcitabine at a fixed dose rate of 10 mg/m(2) per min (1200 mg/m(2), two hours infusion), and carboplatin and plasma gemcitabine concentrations were measured by ion-pair reversed-phase high-performance liquid chromatography (HPLC). 3P97 Pharmaceutical Kinetics Software was used for the calculation of pharmacokinetic parameters. The obtained mean parameters, elimination half life (t(1/2)) (10.67+/-3.38 min), area under the curve (AUC) (7.55+/-1.53 (microg x h)/ml), and clearance (CL) (3940.05+/-672.08 ml/min), were consistent with those reported in literature. The hematologic toxicology result showed that the regimen was effective on and tolerated by the patients. PMID- 15822165 TI - Blood pressure and a B vitamin. PMID- 15822166 TI - Stents don't protect the entire artery. PMID- 15822167 TI - H. Jeremy C. Swan. PMID- 15822168 TI - Patient safety alert. Has the nursing shortage decreased health care quality? PMID- 15822170 TI - Cytochrome P450 activation of arylamines and heterocyclic amines. AB - Arylamines and heterocyclic arylamines (HAAs) are of particular interest because of demonstrated carcinogenicity in animals and humans and the broad exposure to many of these compounds. The activation of these, and also some arylamine drugs, involves N-hydroxylation, usually by cytochrome P450 (P450). P450 1A2 plays a prominent role in these reactions. However, P450 1A1 and 1B1 and other P450s are also important in humans as well as experimental animals. Some arylamines (including drugs) are N-hydroxylated predominantly by P450s other than those in Family 1. Other oxygenases can also have roles. An important issue is extrapolation between species in predicting cancer risks, as shown by the low rates of HAA activation by rat P450 1A2 and low levels of P450 1A2 expression in some nonhuman primates. PMID- 15822171 TI - Glutathione transferases. AB - This review describes the three mammalian glutathione transferase (GST) families, namely cytosolic, mitochondrial, and microsomal GST, the latter now designated MAPEG. Besides detoxifying electrophilic xenobiotics, such as chemical carcinogens, environmental pollutants, and antitumor agents, these transferases inactivate endogenous alpha,beta-unsaturated aldehydes, quinones, epoxides, and hydroperoxides formed as secondary metabolites during oxidative stress. These enzymes are also intimately involved in the biosynthesis of leukotrienes, prostaglandins, testosterone, and progesterone, as well as the degradation of tyrosine. Among their substrates, GSTs conjugate the signaling molecules 15-deoxy delta(12,14)-prostaglandin J2 (15d-PGJ2) and 4-hydroxynonenal with glutathione, and consequently they antagonize expression of genes trans-activated by the peroxisome proliferator-activated receptor gamma (PPARgamma) and nuclear factor erythroid 2 p45-related factor 2 (Nrf2). Through metabolism of 15d-PGJ2, GST may enhance gene expression driven by nuclear factor-kappaB (NF-kappaB). Cytosolic human GST exhibit genetic polymorphisms and this variation can increase susceptibility to carcinogenesis and inflammatory disease. Polymorphisms in human MAPEG are associated with alterations in lung function and increased risk of myocardial infarction and stroke. Targeted disruption of murine genes has demonstrated that cytosolic GST isoenzymes are broadly cytoprotective, whereas MAPEG proteins have proinflammatory activities. Furthermore, knockout of mouse GSTA4 and GSTZ1 leads to overexpression of transferases in the Alpha, Mu, and Pi classes, an observation suggesting they are part of an adaptive mechanism that responds to endogenous chemical cues such as 4-hydroxynonenal and tyrosine degradation products. Consistent with this hypothesis, the promoters of cytosolic GST and MAPEG genes contain antioxidant response elements through which they are transcriptionally activated during exposure to Michael reaction acceptors and oxidative stress. PMID- 15822173 TI - Fat cells: afferent and efferent messages define new approaches to treat obesity. AB - For a long time neural and endocrine messages were studied for their impact on adipocyte metabolism and control of storage/release of fatty acids. In fact, bidirectional communication exists between adipocytes and other tissues. Several molecules secreted from adipocytes are involved in fat cell signaling to other tissues. Adipocyte products could initiate antagonistic effects on target tissues. Fat cells produce peptides that can elicit insulin resistance, such as tumor necrosis factor-alpha and resistin, as well as hormones that can improve insulin resistance, such as leptin and adiponectin. Secretion of complement proteins, proinflammatory cytokines, procoagulant, and acute phase reactant proteins have also been observed in adipocytes. There is much to learn about how these signals function. It is unlikely that all the adipocyte's endocrine and paracrine signals have been identified. Putative pharmacological strategies aiming at modulation of afferent and efferent fat cell messages are reviewed and discussed. PMID- 15822174 TI - The role of metabolic activation in drug-induced hepatotoxicity. AB - The importance of reactive metabolites in the pathogenesis of drug-induced toxicity has been a focus of research interest since pioneering investigations in the 1950s revealed the link between toxic metabolites and chemical carcinogenesis. There is now a great deal of evidence that shows that reactive metabolites are formed from drugs known to cause hepatotoxicity, but how these toxic species initiate and propagate tissue damage is still poorly understood. This review summarizes the evidence for reactive metabolite formation from hepatotoxic drugs, such as acetaminophen, tamoxifen, diclofenac, and troglitazone, and the current hypotheses of how this leads to liver injury. Several hepatic proteins can be modified by reactive metabolites, but this in general equates poorly with the extent of toxicity. Much more important may be the identification of the critical proteins modified by these toxic species and how this alters their function. It is also important to note that the toxicity of reactive metabolites may be mediated by noncovalent binding mechanisms, which may also have profound effects on normal liver physiology. Technological developments in the wake of the genomic revolution now provide unprecedented power to characterize and quantify covalent modification of individual target proteins and their functional consequences; such information should dramatically improve our understanding of drug-induced hepatotoxic reactions. PMID- 15822175 TI - Natural health products and drug disposition. AB - Botanicals such as herbal products (HPs) and nutraceuticals (NCs) are often regarded as low risk because of their long history of human use. Anecdotal and literature reports of adverse drug events (ADEs) and clinical studies with HPs are increasing, but many of the reports are incomplete and contradictory. These reports need to identify confounding factors and explain contradictory findings if they are to help health care professionals or patients understand what risks are involved. HPs are complex botanicals, not single-active ingredient (SAI) products. Studies can be confounded by different manufacturing processes and formulations, including cosmetics and food supplements; environment; chemotypes; misidentification or adulteration; and factors associated with the patient or user population such as use, total drug load, and genetics. Future studies need to be conducted with characterized product that includes all commercially available related products. Clinical trials should be relevant to the user population and take into account the confounding factors that may influence the interpretation of the findings. PMID- 15822176 TI - Biomarkers in psychotropic drug development: integration of data across multiple domains. AB - This review focuses on the current status of biomarkers and/or approaches critical to assessing novel neuroscience targets with an emphasis on new paradigms and challenges in this field of research. The importance of biomarker data integration for psychotropic drug development is illustrated with examples for clinically used medications and investigational drugs. The question remains how to verify access to the brain. Early imaging studies including micro-PET can help to overcome this. However, in case of delayed tracer development or because of no feasible application of brain imaging effects of the molecule, using CSF as a matrix could fill this gap. Proteomic research using CSF will hopefully have a major impact on the development of treatments for psychiatric disorders. PMID- 15822172 TI - Pleiotropic effects of statins. AB - Statins are potent inhibitors of cholesterol biosynthesis. In clinical trials, statins are beneficial in the primary and secondary prevention of coronary heart disease. However, the overall benefits observed with statins appear to be greater than what might be expected from changes in lipid levels alone, suggesting effects beyond cholesterol lowering. Indeed, recent studies indicate that some of the cholesterol-independent or "pleiotropic" effects of statins involve improving endothelial function, enhancing the stability of atherosclerotic plaques, decreasing oxidative stress and inflammation, and inhibiting the thrombogenic response. Furthermore, statins have beneficial extrahepatic effects on the immune system, CNS, and bone. Many of these pleiotropic effects are mediated by inhibition of isoprenoids, which serve as lipid attachments for intracellular signaling molecules. In particular, inhibition of small GTP-binding proteins, Rho, Ras, and Rac, whose proper membrane localization and function are dependent on isoprenylation, may play an important role in mediating the pleiotropic effects of statins. PMID- 15822177 TI - Neonicotinoid insecticide toxicology: mechanisms of selective action. AB - The neonicotinoids, the newest major class of insecticides, have outstanding potency and systemic action for crop protection against piercing-sucking pests, and they are highly effective for flea control on cats and dogs. Their common names are acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, and thiamethoxam. They generally have low toxicity to mammals (acute and chronic), birds, and fish. Biotransformations involve some activation reactions but largely detoxification mechanisms. In contrast to nicotine, epibatidine, and other ammonium or iminium nicotinoids, which are mostly protonated at physiological pH, the neonicotinoids are not protonated and have an electronegative nitro or cyano pharmacophore. Agonist recognition by the nicotinic receptor involves cation-pi interaction for nicotinoids in mammals and possibly a cationic subsite for interaction with the nitro or cyano substituent of neonicotinoids in insects. The low affinity of neonicotinoids for vertebrate relative to insect nicotinic receptors is a major factor in their favorable toxicological profile. PMID- 15822178 TI - Glyceraldehyde-3-phosphate dehydrogenase, apoptosis, and neurodegenerative diseases. AB - Increasing evidence supports the notion that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a protein with multiple functions, including its surprising role in apoptosis. GAPDH is overexpressed and accumulates in the nucleus during apoptosis induced by a variety of insults in diverse cell types. Knockdown of GAPDH using an antisense strategy demonstrates its involvement in the apoptotic cascade in which GAPDH nuclear translocation appears essential. Knowledge concerning the mechanisms underlying GAPDH nuclear translocation and subsequent cell death is growing. Additional evidence suggests that GAPDH may be an intracellular sensor of oxidative stress during early apoptosis. Abnormal expression, nuclear accumulation, changes in physical properties, and loss of glycolytic activity of GAPDH have been found in cellular and transgenic models as well as postmortem tissues of several neurodegenerative diseases. The interaction of GAPDH with disease-related proteins as well as drugs used to treat these diseases suggests that it is a potential molecular target for drug development. PMID- 15822179 TI - Epoxide hydrolases: mechanisms, inhibitor designs, and biological roles. AB - Organisms are exposed to epoxide-containing compounds from both exogenous and endogenous sources. In mammals, the hydration of these compounds by various epoxide hydrolases (EHs) can not only regulate their genotoxicity but also, for lipid-derived epoxides, their endogenous roles as chemical mediators. Recent findings suggest that the EHs as a family represent novel drug discovery targets for regulation of blood pressure, inflammation, cancer progression, and the onset of several other diseases. Knowledge of the EH mechanism provides a solid foundation for the rational design of inhibitors, and this review summarizes the current understanding of the catalytic mechanism of the EHs. Although the overall EH mechanism is now known, the molecular basis of substrate selectivity, possible allosteric regulation, and many fine details of the catalytic mechanism remain to be solved. Finally, recent development in the design of EH inhibitors and the EH biological role are discussed. PMID- 15822180 TI - Nitroxyl (HNO): chemistry, biochemistry, and pharmacology. AB - Recent discoveries of novel and potentially important biological activity have spurred interest in the chemistry and biochemistry of nitroxyl (HNO). It has become clear that, among all the nitrogen oxides, HNO is unique in its chemistry and biology. Currently, the intimate chemical details of the biological actions of HNO are not well understood. Moreover, many of the previously accepted chemical properties of HNO have been recently revised, thus requiring reevaluation of possible mechanisms of biological action. Herein, we review these developments in HNO chemistry and biology. PMID- 15822181 TI - Tyrosine kinase inhibitors and the dawn of molecular cancer therapeutics. AB - The clinical application of tyrosine kinase inhibitors for cancer treatment represents a therapeutic breakthrough. The rationale for developing these compounds rests on the observation that tyrosine kinase enzymes are critical components of the cellular signaling apparatus and are regularly mutated or otherwise deregulated in human malignancies. Novel tyrosine kinase inhibitors are designed to exploit the molecular differences between tumor cells and normal tissues. Herein, we will review the current state-of-the-art using agents that target as prototypes Bcr-Abl, platelet-derived growth factor receptor (PDGFR), KIT (stem cell factor receptor), and epidermal growth factor receptor (EGFR). These compounds are remarkably effective in treating diverse cancers that are highly resistant to conventional treatment, including various forms of leukemia, hypereosinophilic syndrome, mast cell disease, sarcomas, and lung cancer. It is now clear that the molecular defects underlying cancer can be targeted with designer drugs that yield striking salutary effects with minimal toxicity. PMID- 15822182 TI - Actions of adenosine at its receptors in the CNS: insights from knockouts and drugs. AB - Adenosine and its receptors have been the topic of many recent reviews. These reviews provide a good summary of much of the relevant literature--including the older literature. We have, therefore, chosen to focus the present review on the insights gained from recent studies on genetically modified mice, particularly with respect to the function of adenosine receptors and their potential as therapeutic targets. The information gained from studies of drug effects is discussed in this context, and discrepancies between genetic and pharmacological results are highlighted. PMID- 15822183 TI - Regulation and inhibition of arachidonic acid omega-hydroxylases and 20-HETE formation. AB - Cytochrome P450-catalyzed metabolism of arachidonic acid is an important pathway for the formation of paracrine and autocrine mediators of numerous biological effects. The omega-hydroxylation of arachidonic acid generates significant levels of 20-hydroxyeicosatetraenoic acid (20-HETE) in numerous tissues, particularly the vasculature and kidney tubules. Members of the cytochrome P450 4A and 4F families are the major omega-hydroxylases, and the substrate selectivity and regulation of these enzymes has been the subject of numerous studies. Altered expression and function of arachidonic acid omega-hydroxylases in models of hypertension, diabetes, inflammation, and pregnancy suggest that 20-HETE may be involved in the pathogenesis of these diseases. Our understanding of the biological significance of 20-HETE has been greatly aided by the development and characterization of selective and potent inhibitors of the arachidonic acid omega hydroxylases. This review discusses the substrate selectivity and expression of arachidonic acid omega-hydroxylases, regulation of these enzymes during disease, and the application of enzyme inhibitors to study 20-HETE function. PMID- 15822184 TI - Cytochrome P450 ubiquitination: branding for the proteolytic slaughter? AB - The hepatic cytochromes P450 (P450s) are monotopic endoplasmic reticulum (ER) anchored hemoproteins engaged in the enzymatic oxidation of a wide variety of endo- and xenobiotics. In the course of these reactions, the enzymes generate reactive O(2) species and/or reactive metabolic products that can attack the P450 heme and/or protein moiety and structurally and functionally damage the enzyme. The in vivo conformational unraveling of such a structurally damaged P450 signals its rapid removal via the cellular sanitation system responsible for the proteolytic disposal of structurally aberrant, abnormal, and/or otherwise malformed proteins. A key player in this process is the ubiquitin (Ub)-dependent 26S proteasome system. Accordingly, the structurally deformed P450 protein is first branded for recognition and proteolytic removal by the 26S proteasome with an enzymatically incorporated polyUb tag. P450s of the 3A subfamily such as the major human liver enzyme CYP3A4 are notorious targets for this process, and they represent excellent prototypes for the understanding of integral ER protein ubiquitination. Not all the participants in hepatic CYP3A ubiquitination and subsequent proteolytic degradation have been identified. The following discussion thus addresses the various known and plausible events and/or cellular participants involved in this multienzymatic P450 ubiquitination cascade, on the basis of our current knowledge of other eukaryotic models. In addition, because the detection of ubiquitinated P450s is technically challenging, the critical importance of appropriate methodology is also discussed. PMID- 15822185 TI - Proteasome inhibition in multiple myeloma: therapeutic implication. AB - Normal cellular functioning requires processing of proteins regulating cell cycle, growth, and apoptosis. The ubiquitin-proteasome pathway (UBP) modulates intracellular protein degradation. Specifically, the 26S proteasome is a multienzyme protease that degrades misfolded or redundant proteins; conversely, blockade of the proteasomal degradation pathways results in accumulation of unwanted proteins and cell death. Because cancer cells are more highly proliferative than normal cells, their rate of protein translation and degradation is also higher. This notion led to the development of proteasome inhibitors as therapeutics in cancer. The FDA recently approved the first proteasome inhibitor bortezomib (Velcade), formerly known as PS-341, for the treatment of newly diagnosed and relapsed/refractory multiple myeloma (MM). Ongoing studies are examining other novel proteasome inhibitors, in addition to bortezomib, for the treatment of MM and other cancers. PMID- 15822186 TI - Clinical and toxicological relevance of CYP2C9: drug-drug interactions and pharmacogenetics. AB - CYP2C9 is a major cytochrome P450 enzyme that is involved in the metabolic clearance of a wide variety of therapeutic agents, including nonsteroidal antiinflammatories, oral anticoagulants, and oral hypoglycemics. Disruption of CYP2C9 activity by metabolic inhibition or pharmacogenetic variability underlies many of the adverse drug reactions that are associated with the enzyme. CYP2C9 is also the first human P450 to be crystallized, and the structural basis for its substrate and inhibitor selectivity is becoming increasingly clear. New, ultrapotent inhibitors of CYP2C9 have been synthesised that aid in the development of quantitative structure-activity relationship (QSAR) models to facilitate drug redesign, and extensive resequencing of the gene and studies of its regulation will undoubtedly help us understand interindividual variability in drug response and toxicity controlled by this enzyme. PMID- 15822187 TI - Clinical development of histone deacetylase inhibitors as anticancer agents. AB - Acetylation is a key posttranslational modification of many proteins responsible for regulating critical intracellular pathways. Although histones are the most thoroughly studied of acetylated protein substrates, histone acetyltransferases (HATs) and deacetylases (HDACs) are also responsible for modifying the activity of diverse types of nonhistone proteins, including transcription factors and signal transduction mediators. HDACs have emerged as uncredentialed molecular targets for the development of enzymatic inhibitors to treat human cancer, and six structurally distinct drug classes have been identified with in vivo bioavailability and intracellular capability to inhibit many of the known mammalian members representing the two general types of NAD+-independent yeast HDACs, Rpd3 (HDACs 1, 2, 3, 8) and Hda1 (HDACs 4, 5, 6, 7, 9a, 9b, 10). Initial clinical trials indicate that HDAC inhibitors from several different structural classes are very well tolerated and exhibit clinical activity against a variety of human malignancies; however, the molecular basis for their anticancer selectivity remains largely unknown. HDAC inhibitors have also shown preclinical promise when combined with other therapeutic agents, and innovative drug delivery strategies, including liposome encapsulation, may further enhance their clinical development and anticancer potential. An improved understanding of the mechanistic role of specific HDACs in human tumorigenesis, as well as the identification of more specific HDAC inhibitors, will likely accelerate the clinical development and broaden the future scope and utility of HDAC inhibitors for cancer treatment. PMID- 15822188 TI - The magic bullets and tuberculosis drug targets. AB - Modern chemotherapy has played a major role in our control of tuberculosis. Yet tuberculosis still remains a leading infectious disease worldwide, largely owing to persistence of tubercle bacillus and inadequacy of the current chemotherapy. The increasing emergence of drug-resistant tuberculosis along with the HIV pandemic threatens disease control and highlights both the need to understand how our current drugs work and the need to develop new and more effective drugs. This review provides a brief historical account of tuberculosis drugs, examines the problem of current chemotherapy, discusses the targets of current tuberculosis drugs, focuses on some promising new drug candidates, and proposes a range of novel drug targets for intervention. Finally, this review addresses the problem of conventional drug screens based on inhibition of replicating bacilli and the challenge to develop drugs that target nonreplicating persistent bacilli. A new generation of drugs that target persistent bacilli is needed for more effective treatment of tuberculosis. PMID- 15822189 TI - Molecular mechanisms of resistance in antimalarial chemotherapy: the unmet challenge. AB - The enormous public health problem posed by malaria has been substantially worsened in recent years by the emergence and worldwide spread of drug-resistant parasites. The utility of two major therapies, chloroquine and the synergistic combination of pyrimethamine/sulfadoxine, is now seriously compromised. Although several genetic mechanisms have been described, the major source of drug resistance appears to be point mutations in protein target genes. Clinically significant resistance to these agents requires the accumulation of multiple mutations, which genetic studies of parasite populations suggest arise focally and sweep through the population. Efforts to circumvent resistance range from the use of combination therapy with existing agents to laboratory studies directed toward discovering novel targets and therapies. PMID- 15822190 TI - Signaling networks in living cells. AB - Recent advances in cell signaling research suggest that multiple sets of signal transducing molecules are preorganized and sequestered in distinct compartments within the cell. These compartments are assembled and maintained by specific cellular machinery. The molecular ecology within a compartment creates an environment that favors the efficient and accurate integration of signaling information arriving from humoral, mechanical, and nutritional sources. The functional organization of these compartments suggests they are the location of signaling networks that naturally organize into hierarchical interconnected sets of molecules through their participation in different classes of interacting units. An important goal is to determine the contribution of the compartment to the function of these networks in living cells. PMID- 15822191 TI - Aberrant DNA methylation as a cancer-inducing mechanism. AB - Aberrant DNA methylation is the most common molecular lesion of the cancer cell. Neither gene mutations (nucleotide changes, deletions, recombinations) nor cytogenetic abnormalities are as common in human tumors as DNA methylation alterations. The most studied change of DNA methylation in neoplasms is the silencing of tumor suppressor genes by CpG island promoter hypermethylation, which targets genes such as p16(INK4a), BRCA1, and hMLH1. There is a profile of CpG island hypermethylation according to the tumor type, and genes silent by methylation represent all cellular pathways. The introduction of bisulfite-PCR methodologies combined with new genomic approaches provides a comprehensive spectrum of the genes undergoing this epigenetic change across all malignancies. However, we still know very little about how this aberrant DNA methylation "invades" the previously unmethylated CpG island and how it is maintained through cell divisions. Furthermore, we should remember that this methylation occurs in the context of a global genomic loss of 5-methylcytosine (5mC). Initial clues to understand this paradox should be revealed from the current studies of DNA methyltransferases and methyl CpG binding proteins. From the translational standpoint, we should make an effort to validate the use of some hypermethylated genes as biomarkers of the disease; for example, it may occur with MGMT and GSTP1 in brain and prostate tumors, respectively. Finally, we must expect the development of new and more specific DNA demethylating agents that awake these methyl-dormant tumor suppressor genes and prove their therapeutic values. The expectations are high. PMID- 15822192 TI - The cardiac fibroblast: therapeutic target in myocardial remodeling and failure. AB - Cardiac fibroblasts play a central role in the maintenance of extracellular matrix in the normal heart and as mediators of inflammatory and fibrotic myocardial remodeling in the injured and failing heart. In this review, we evaluate the cardiac fibroblast as a therapeutic target in heart disease. Unique features of cardiac fibroblast cell biology are discussed in relation to normal and pathophysiological cardiac function. The contribution of cardiac fibrosis as an independent risk factor in the outcome of heart failure is considered. Candidate drug therapies that derive benefit from actions on cardiac fibroblasts are summarized, including inhibitors of angiotensin-aldosterone systems, endothelin receptor antagonists, statins, anticytokine therapies, matrix metalloproteinase inhibitors, and novel antifibrotic/anti-inflammatory agents. These findings point the way to future challenges in cardiac fibroblast biology and pharmacotherapy. PMID- 15822193 TI - Evaluation of drug-drug interaction in the hepatobiliary and renal transport of drugs. AB - Recent studies have revealed the import role played by transporters in the renal and hepatobiliary excretion of many drugs. These transporters exhibit a broad substrate specificity with a degree of overlap, suggesting the possibility of transporter-mediated drug-drug interactions with other substrates. This review is an overview of the roles of transporters and the possibility of transporter mediated drug-drug interactions. Among the large number of transporters, we compare the Ki values of inhibitors for organic anion transporting polypeptides (OATPs) and organic anion transporters (OATs) and their therapeutic unbound concentrations. Among them, cephalosporins and probenecid have the potential to produce clinically relevant OAT-mediated drug-drug interactions, whereas cyclosporin A and rifampicin may trigger OATP-mediated ones. These drugs have been reported to cause drug-drug interactions in vivo with OATs or OATP substrates, suggesting the possibility of transporter-mediated drug-drug interactions. To avoid adverse consequences of such transporter-mediated drug drug interactions, we need to be more aware of the role played by drug transporters as well as those caused by drug metabolizing enzymes. PMID- 15822194 TI - Dual specificity protein phosphatases: therapeutic targets for cancer and Alzheimer's disease. AB - The complete sequencing of the human genome is generating many novel targets for drug discovery. Understanding the pathophysiological roles of these putative targets and assessing their suitability for therapeutic intervention has become the major hurdle for drug discovery efforts. The dual-specificity phosphatases (DSPases), which dephosphorylate serine, threonine, and tyrosine residues in the same protein substrate, have important roles in multiple signaling pathways and appear to be deregulated in cancer and Alzheimer's disease. We examine the potential of DSPases as new molecular therapeutic targets for the treatment of human disease. PMID- 15822204 TI - The evolution of male traits in social insects. AB - Pair formation in social insects mostly happens early in adult life and away from the social colony context, which precludes promiscuity in the usual sense. Termite males have continuous sperm production, but males of social Hymenoptera have fixed complements of sperm, except for a few species that mate before female dispersal and show male-fighting and lifelong sperm production. We develop an evolutionary framework for testing sexual selection and sperm competition theory across the advanced eusocial insects (ants, wasps, bees, termites) and highlight two areas related to premating sexual selection (sexual dimorphism and male mate number) that have remained understudied and in which considerable progress can be achieved with relatively simple approaches. We also infer that mating plugs may be relatively common, and we review further possibilities for postmating sexual selection, which gradually become less likely in termite evolution, but for which eusocial Hymenoptera provide unusual opportunities because they have clonal ejaculates and store viable sperm for up to several decades. PMID- 15822205 TI - Ecology of interactions between weeds and arthropods. AB - Weeds and arthropods interact in agricultural systems. Weeds can directly serve as food sources or provide other ecosystem resources for herbivorous arthropods, and indirectly serve carnivorous (beneficial) arthropods by providing food and shelter to their prey. Weeds can serve as alternative hosts for pest and beneficial arthropods when their preferred crop host is absent. Herbivory on crops by pest arthropods reduces the competitive ability of crop plants, leading to increased weed growth. Interactions between weeds and arthropods have several implications to integrated pest management (IPM). Pest and beneficial arthropod populations can be maintained in the absence of crop hosts. This statement also applies to all other pests that use weeds as a food source, including pathogens, nematodes, mollusks, and vertebrates. Weeds outside crop fields that maintain overwintering populations of arthropod pests are the major reason for the development of area-wide IPM programs for certain mobile arthropod pests. Weeds can serve as a source of increased diversity in agroecosystems. Increased diversity has been the rationale for enhancing biological control of arthropod pests through habitat management. The consequences of such approaches are difficult to predict on a multispecies IPM basis. PMID- 15822206 TI - Cloning and molecular characterization of the first aquatic hyaluronidase, SFHYA1, from the venom of stonefish (Synanceja horrida). AB - We report here for the first time the molecular characterization of a hyaluronidase from an aquatic source. SFHYA1 is the hyaluronidase found in the venom gland of stonefish, Synanceja horrida. Using a cDNA segment amplified with degenerate oligonucleotides based on the amino acid sequences of a conserved region in testicular-type hyaluronidases and a tryptic fragment of SFHYA1, clones encoding the precursor of this enzyme were isolated from a cDNA library prepared from stonefish venom glands. The deduced amino acid sequence of SFHYA1 shows that SFHYA1 is expressed as a precursor peptide with a 28-residue signal peptide for targeting it into endoplasmic reticulum. Mature SFHYA1 is a polypeptide composed of 449 residues containing three potential N-glycosylation sites, four putative hyaluronan-binding motifs [B(X)7B] and various residues implicated in substrate binding and catalysis. This cDNA was expressed in an active form in insect-cells but not in E. coli. Homology-based computational analyses suggested that SFHYA1 closely resembles the PH-20 family of hyaluronidases. PMID- 15822208 TI - Obesity in children--what might be its impact on life expectancy? PMID- 15822207 TI - Morphine pharmacokinetics following intra-articular administration of a novel sustained release opioid (CDS-PM-101) for the relief of post-operative orthopaedic pain. PMID- 15822209 TI - Resources. "Allergic asthma for dummies"...and a Web-based tool to improve immunizations and health screenings. PMID- 15822210 TI - Water and infections: what we drink...and where we swim. PMID- 15822211 TI - A timely reminder: infections from pet turtles. PMID- 15822213 TI - Product recalls. Recall: Flashing Pacifier. PMID- 15822212 TI - Child safety. Mary Ucci: getting the playground ready for spring. PMID- 15822214 TI - A technique for emergency liver packing. AB - Control of liver hemorrhage may present a daunting clinical scenario. Use of liver packing techniques is highly effective to control bleeding but can result in significant recurrent bleeding with pack removal. Such bleeding is particularly a problem when large portions of the hepatic parenchymal surface and Glisson's capsule have been disrupted. We describe, herein, our approach to hepatic packing in scenarios where a large component of hepatic capsular disruption has occurred. Use of a non-stick bowel bag is employed on the disrupted liver surface, which, when removed, will not result in liver rebleeding. This technique has been used successfully in the management of five cases of severe liver injury with extensive capsular disruption. Familiarity with such an approach may facilitate management of similar liver injuries. PMID- 15822215 TI - Complex coacervation with heparin results in decreased lysozyme stability. PMID- 15822216 TI - Nano-particulate systems and other novel concepts in drug delivery. Bubble assisted drug delivery. PMID- 15822217 TI - [Usutu virus in Austria reminds one of the West Nile virus in the USA]. PMID- 15822218 TI - Qualitative prediction of solubilization of highly hydrophobic drugs in block copolymer micelles. PMID- 15822219 TI - Factorial design, physicochemical characterisation and activity of ciprofloxacin loaded PLGA nanoparticles for ocular use. PMID- 15822220 TI - Development and evaluation of sustained release ocular minitablets. PMID- 15822221 TI - Seizure during alpha-interferon therapy. PMID- 15822222 TI - A sensory valve in liposomal drug delivery systems. PMID- 15822223 TI - Oligopeptides as targeting structures in cancer theraPY. PMID- 15822225 TI - Conference reports. PMID- 15822224 TI - Micelles from new biodegradable amphiphilic block copolymers containing PEG AND PCL. PMID- 15822226 TI - [Orthostatic distal angiohypertension of the lower extremities]. PMID- 15822227 TI - [I.P. Pavlov--the author of experimental surgery of the stomach. (To the 100th anniversary of I. P. Pavlov's Nobel Prize Award)]. PMID- 15822228 TI - Channel protein-containing liposomes as delivery vehicles for the controlled release of drugs-optimization of the lipid composition. PMID- 15822229 TI - Thermosensitive and biodegradable polymeric micelles with transient stability. PMID- 15822230 TI - PEG-phospholipid micelles for the delivery of amphotericin B. PMID- 15822231 TI - Controlled drug delivery with ultrasound and gas microbubbles. PMID- 15822232 TI - PTMC and MPEG-PTMC microparticles for hydrophilic drug delivery. PMID- 15822233 TI - Covalent attachment of an NLS-peptide to linear dna does not enhance transfection efficiency of cationic polymer based gene delivery systems. PMID- 15822234 TI - Antisense peptide nucleic acid delivered by core-shell microspheres. PMID- 15822235 TI - Generation of LTB-based mucosal vaccines and production in plants. PMID- 15822236 TI - In-vivo delivery of DNA and protein using conceptually new cationic, sunfish', amphiphiles. PMID- 15822237 TI - Studies on the intracellular release of genetic drugs from pharmaceutical carriers. PMID- 15822238 TI - Structurally well-defined copolymers of poly(ethylene glycol) and low molecular weight linear polyethylenimine as vectors for gene delivery. PMID- 15822240 TI - DOE medical research funding cut in FY 2006 budget; SNM calls for nuclear medicine community action. PMID- 15822239 TI - Bridging the gap between research and practice in bereavement: report from the Center for the Advancement of Health. AB - This article discusses issues in bridging the gap between research and practice in the field of bereavement. A conceptual framework is developed that emphasizes that the gulf reflects the lack of exchange and mutual influence between researchers and practitioners. Studies are presented using qualitative and quantitative methods to gain a better understanding of the nature of the gap between research and practice in bereavement. Finally recommendations are made for practitioners, researchers, and organizations to take steps toward more thoroughly integrating bereavement research and practice for the advancement of both. PMID- 15822241 TI - Scholarship, students, and practitioners: bringing scholarship into the expectations of practitioners. AB - Increased attention is being given to bridging the gap separating thanatology researchers and practitioners. College undergraduates studying to be human service professionals represent a cohort with a significant stake in bridging this gap. Inculcating in college students expectations as well as expertise to use research as practitioners not only makes sense but also is an ethical obligation for college faculty. Two noteworthy models of academic practice Boyer's vision of scholarship and Schon's idea of reflective practice-provide a conceptual framework for faculty engaging practitioners-in-the-making in the use of research. An example of one effort to engage students in such learning is offered, along with discussion of some implications and alternatives. PMID- 15822242 TI - "False positive" claims of near-death experiences and "false negative" denials of near-death experiences. AB - Some persons who claim to have had near-death experiences (NDEs) fail research criteria for having had NDEs ("false positives"); others who deny having had NDEs do meet research criteria for having had NDEs ("false negatives"). The author evaluated false positive claims and false negative denials in an organization that promotes near-death research and in psychiatric outpatients. The frequency of false positives and negatives varied in samples that differed in prevalence of, and knowledge about, NDEs. The influence of participants' knowledge about NDEs on the findings of near-death research makes it critically important to use standardized criteria for identifying NDEs. PMID- 15822243 TI - Anxiety and death anxiety in Egyptian and Spanish nursing students. AB - Two samples of female nursing undergraduates from Egypt (n = 132) and Spain (n = 126) responded to the Arabic Scale of Death Anxiety, the Spanish Death Anxiety Inventory, the Templer's Death Anxiety Scale, the Kuwait University Anxiety Scale, and the State-Trait Anxiety Inventory-Trait Subscale. Each sample answered the scales in their native language. Alpha reliabilities of the total score scales ranged from 0.65 to 0.91 (Egyptian sample) and between 0.71 and 0.90 (Spanish sample). The Spanish respondents attained significantly lower mean scores than the Egyptian sample in all the 5 scales. All the intercorrelations between these scales were statistically significant, and yielded two factors: Death Anxiety and General Anxiety in both countries. The correlations between these factors were significant, positive, and moderate, that is, 0.57 and 0.50 in the Egyptian and Spanish samples, respectively. The general conclusion is that Death Anxiety and General Anxiety are 2 different, but correlated factors. PMID- 15822244 TI - Death anxiety and personal growth in adolescents experiencing the death of a grandparent. AB - Relationships between bereavement following the death of a grandparent and adolescent death anxiety levels were examined using Hogan's Inventory of Bereavement and the revised Death Anxiety Scale within private schools. Of the 226 adolescent respondents (aged 11-18) who completed questionnaires, 124 had experienced the death of a grandparent. The regression model indicated that grief due to bereavement was found to be significantly associated with death anxiety. The factors of personal growth, gender, time since death, and numbers of deaths experienced were not found to be significant predictors within the model. The findings encourage further exploration into the nature of the grandparent adolescent relationship as well as the impact of type of death experienced. PMID- 15822245 TI - Inhalational anthrax with acute respiratory distress syndrome. AB - A 10-year-old boy presented with facial swelling, thick exudates in the nasal cavities with membranes covering the nasopharynx, shock and respiratory distress. X-ray of the paranasal sinuses showed opacification of both maxillary sinuses. Nasal diphtheria was suspected but culture of the membranes grew Bacillus anthracis. Chest X-ray showed mediastinal widening and extensive pulmonary infiltrates compatible with respiratory anthrax. PMID- 15822246 TI - Sanford Louis Palay (23 September 1918 - 5 August 2002). PMID- 15822247 TI - Owsei Temkin (6 October 1902 - 18 July 2002). PMID- 15822248 TI - Public culture and public understanding of genetics: a focus group study. AB - As the role of genetic science in everyday life has grown, policymakers have become concerned about Americans' understandings of this science. Much effort has been devoted to formal schooling, but less attention has been paid to the role of public culture in shaping public understanding of genetics. Research into public cultural messages about genetics has claimed that the public is likely to adopt problematic accounts, but few studies have explored the public's articulation of these messages. This study is based on 25 focus groups convened to explore the lay public's understanding of genetics. The study found that the public processed a greater variety of messages than assumed by previous researchers, including documentaries, non-science-fiction films, and popular television in addition to previous researchers' focus on science fiction and news media. The study also found that the public does not process the messages through the linear, transmission model assumed by previous research. The public processes messages about genetics complexly and critically. On the basis of these findings, the study suggests that researchers should include a greater variety of texts about genetics in their research and attend more fully to audience processing in addition to content analyses of these texts. PMID- 15822249 TI - Medical history for the medical student. AB - The modern medical student is necessarily heavily burdened with instruction in Medical Science. When the teaching of Medical History is added to the university course, it is first necessary to stimulate student interest in that discipline and show its clinical relevance. PMID- 15822250 TI - Renkioi: a forgotten Crimean War hospital and its significance. AB - Renkioi Civil Hospital was built late in the Crimean War (1854-6) in Turkey on the Dardanelles. Designed by Isambard Kingdom Brunel, its prefabricated structure was a brilliant engineering innovation. As a civil hospital for military patients, it was staffed by experienced civilian doctors, thereby relieving the shortage of military doctors. Renkioi is remembered as an astonishing early prefabricated structure. However the war was soon to end and it was never used to near capacity. Thus, its other successful features are largely forgotten. It demonstrated the advantages of a doctors, rather than a military officer, being in complete command of a hospital and this was later accepted by the army. Renkioi also showed how infection could be reduced by able staff in a well administered, properly designed hospital with good sanitation. After the war, Dr. Edmund Parkes, its Medical Superintendent, became the first Professor of Hygiene at the new Army Medical School, ensuring that "the prevention of disease and the promotion of health" became the first function of the Army Medical Services. PMID- 15822251 TI - The physician Romolo Spezioli (1642-1723) and his private library in the Public Library of Fermo. AB - The Public Library of Fermo in the Italian Marches, houses an almost unknown treasure for historians of medicine, the personal library of Romolo Spezioli. Spezioli was born in Fermo in 1642 and died in Rome in 1723. He was the personal physician of Cardinal Decio Azzolini junior, Queen Christina of Sweden and Pope Alexander VIII. There are almost 12,000 books, dating from the sixteenth to the early eighteenth centuries, almost all of them on medicine or science. They are housed in the Globe Room, whose creation was made possible by the generosity of Cardinal Azzolini. The library was opened to the public in 1688. PMID- 15822252 TI - Disease in Africa: a medical perspective from the 1950s. AB - So much attention is focused on the current HIV/AIDS epidemic in Africa that there is a tendency to overlook the grievous burden of disease from which the peoples of that Continent have suffered for centuries. This paper, based on letters sent in 1953/54 by a young doctor in Sierra Leone to his parents in Scotland, together with extracts from Makeni Hospital records of the same period, provides a factual account of front-line medicine in West Africa half a century ago. PMID- 15822253 TI - [Georges Dieulafoy (1839-1911) and the teaching of medicine in Paris at the hinge of the Second Empire and the Third Republic]. AB - Armand Trousseau's favourite disciple, Georges Dieulafoy (1839-1911), proved a worthy successor to his master. Although he was an obviously proud person, this did not detract from his outstanding medical merit. He was Professor of Pathology at the Faculty of Medicine in Paris and can be regarded as an outstanding exponent of medical teaching, at that phase of transition between the Second Empire and the Third Republic. The work that he left, in particular the "Manuel de Pathologie Interne", are still easy to read today. PMID- 15822254 TI - [The calculation of Einstein spontaneous emission coefficients of HBr molecule]. AB - The Einstein spontaneous emission coefficients of some rovibrational transitions of HBr molecule have been calculated from the first principle. The calculation is carried out for the deltav = 1 transitions of v < or = 7, J < or = 14 of R and P branches. A time-resolved Fourier transform infrared emission spectrum of the HBr molecule was recorded. The observed intensities of the spectral lines well agree the calculated coefficients. PMID- 15822255 TI - [Luminescence character of Ce3+ in rare earth borate glasses]. AB - Ce3+ doped rare earth borate glasses (BLBL:Ce3+) were synthesized. The excitation and emission spectra and fluorescence decay curves of BLBL:Ce3+ were measured at room temperature. In BLBL:Ce3+ glasses, Ce3+ emits strong purplish-blue light. The results of analyzing spectra and fluorescence lifetime show that there are two kinds of Ce3+ luminescence centres in this kind of rare earth borate glasses. Ce3+ doped rare-earth glasses is a kind of beneficial supplement for Ce3+ fluorescence glasses system. PMID- 15822256 TI - [The absorption spectra of crystal thin film Cs3Cu2I5]. AB - In this paper we measured the absorption spectra of crystal thin film Cs3Cu2I5 at room and liquid nitrogen temperatures, respectively, and calculated the excitonic coefficient of the material. The excitonic coupling energy deltaE(ex)(1) = (0.53 +/- 0.07) eV, excitonic radius a(ex) = 0. 326 nm,and forbidden zone width E(g) = (5.00 +/- 0.07) eV. The spectral analysis shows that the electron and exciton excitations of the Cs3Cu2I5 localize in the CuI subcrystal lattice, and reveals the law that governs the variance of the first excitonic peak position of the serial Cs(x)Cu(1-x) I compounds with their mol-components at low temperature. PMID- 15822257 TI - [The effect of surface structure on the photoluminescence of SnO2 nanoparticles in hydrosols and organosols]. AB - In this paper, we report the optical properties of SnO2 semiconductor nanoparticles in hydrosols and those of SnO2 semiconductor nanoparticles in organosols in which the surfaces of the particles are coated by a layer of organic surfactant molecules. The photoluminescence spectra of SnO2 semiconductor nanoparticles in the hydrosols and organosols in different conditions were measured and discussed. We conclude that the surface structure of the SnO2 semiconductor nanoparticles affects their optical properties strongly. The oxygen deficiencies on the surface of SnO2 semiconductor nanoparticles play an important role in the optical properties. The surface modification of the particles effectively removes the surface defects of the particles and enhances the intensity of luminescence. PMID- 15822258 TI - [Direct upconversion sensitization luminescence of Tm(0.1) Yb (10.9) oxyfluoride vitroceramincs]. AB - This paper studied the direct upconversion sensitization luminescence of Tm (0.1) Yb (10.9) oxyfluoride vitroceramics pumped by 966 nm diode laser for the first time. We found that there are both strong 477 nm three-photon upconversion fluorescence of 1G4 --> 3H6 transition and 799.5 nm two-photon upconversion fluorescence of 3H4 --> 3H6 transition as well as weak upconversion fluorescence of 1D2 --> 3H6, 1Dz --> 3F4,1G4 --> 3F4 and 3F3 --> 3H6 transitions at 361 nm, 449.5 nm, 647.0 nm and (679.5, 698.5 nm), respectively. PMID- 15822260 TI - [The analysis of manganese through electrochemical stripping ellipsometry]. AB - This paper introducing the cathodic stripping ellipsometry with a new function Vop for analyzing manganese quantitatively. The results indicated that the related average deviation of this method are more smaller than the stripping voltamimetry of electrochemistry and providing a lot of information about electrode processes. PMID- 15822259 TI - [Study on laser flash spectroscopy of processes of photoionization of tyrosine: pH effects]. AB - The processes of photoionization of tyrosine on three different pH value solutions were investigated by 248 nm laser flash photolysis. The results showed that the processes of photoionization of tyrosine were affected by pH value of the solution. With pH value increasing, the molecule number of photoionization increases via a monophotonic process, and decreases via a biphotonic process. Furthermore, the pH effect of photoionization of tyrosine resulting from deprotonation of phenolic hydroxyl group in alkaline solution was demonstrated. PMID- 15822261 TI - [Studies on the infrared spectra of mico2-OR or micro2-SR-bridged double-cubane like clusters containing MoFe3S4 cores]. AB - The IR spectra of micro2-OR or micro2-SR- bridged double-cubane-like clusters containing MoFe3S4 cores (Et4N)3 [Mo2Fe6S8Cl6 (OMe)3] (1), (Et4N) [Mo2Fe6S8 (SPh)6 (OMe)3 (2), (Bu4N)3 Mo2Fe6S (SPh)6 (OMe)3] (3), (Et4N)3 [Mo2Fe6S8 (SBu(t))6(OMe)3](4), (Et4N)3[Mo2Fe6S8 (SPh)9](5) have been investigated. Their characteristic frequencies (v)Mo-micro2O(R), (v)Mo-micro2S(R), (v)Mo-S, (v)Fe-S*, (v)Fe-SR, (v)Fe-Cl were assigned by comparing their vibrational frequencies with their structural parameters and by comparing the IR spectra of them with that of the linear clusters. The electrophilic induced effects in different way and the relationship between deltav (i.e. (v)Mo-S* (v)Fe-S*) and Fe --> Mo charge transfer have been discussed. Finaly, the sameness and difference between the double-cubane-like and linear Mo-Fe-S clusters were demonstrated. PMID- 15822262 TI - [Surface-enhanced Raman spectra studies on a novel dipyridophenazine complexes of ruthenium(II) and it's effect on DNA]. AB - SERS is better method at analysing the samples which are prone to emit fluorescence or photodecomposition than other methods. Raman scattering of a novel complexes of Ruthenium (II): Ru(bpy)2dppz and it's effects on DNA were studied with surface-enhanced Raman spectra(SERS). We find that when this complex binds to DNA, only the ligand dppz intercalate into the double helix of DNA, and form a larger conjugate system, affect the metal-to-ligand charge transfer(MLCT), produce strong photoluminescence. All these cause shifting of some Raman peaks of the complex and changex of the relative intensity of some other peaks. And on the charge absorbed spectra, after the addition of DNA, the compex' absorbed peaks of Ru(bpy)2dppz solvent shift to long wavelength, this is also one of the character of the intercalation. PMID- 15822263 TI - [A quantitative study on secondary structure of proteins by FT-Raman spectroscopy]. AB - In this work, proteins have been scanned by FT-Raman spectrometer. Deconvolved spectra, second derivative or fourth derivative spectra were employed to determine positions of individual components. Curve-fitting procedures were carried out to amide I band of original spectra. The broad band was resolved into several components. The resolved band components were assigned to specific protein conformations. The areas of individual components are used to estimate the percentage of the relative secondary structure. Results are in reasonable agreement with data obtained by other methods. PMID- 15822264 TI - [Photoluminescence and UV-VIS absorption spectra characteristics of complexes of ruthenium (II) with 1,10-phenanthroline and 4,7-diphnyl-1,10-phenanthroline]. AB - The preparation and spectroscopic characteristics of the tris (1,10 phenanthroline) and tris (4,7-diphenyl-1,10-phenalthroline) complex ions of rathenium (II) are reported. Under the coordinate field with the property of D3 group, the splitting of the 4d state of Ru2+ is discussed. The transitions which give rise to the UV-VIS absorption spectra and emission spectra of the complexes are assigned. The influence of the coordinate substance on the absorptivity and red-shift is discussed as well. PMID- 15822265 TI - [Spectroscopic study on the structure and orientation of 1-hydroxyl-5-dodecyloxy naphthalene LB films]. AB - Ultraviolet-visible (UV-Vis), infrared transmission reflection absorption (RA) and ATR spectra were used to study the mono-and multi-layer Langmuir-Blodgett (LB) films of C12H25ONaph (1,5)OH at various temperatures to investigate dependencies of the aggregation and phase transitions on the number of monolayers. The UV-vis spectra of the LB films revealed that C12H25ONaph (1,5)OH forms aggregates in the LB film. From the temperature dependence of UV-vis absorption spectra, it is clear that the aggregate was gradually converted to monomer with the temperature increased. A comparison of the infrared transmission and ATR spectra reveals that the hydrocarbon chains were tilted considerably in the LB film. PMID- 15822266 TI - [Forecasting octane numbers of gasoline by NIR spectroscopy and the manufacture of octane number analyzer]. AB - The predictive model of octane numbers, of gasoline was developed by using MLR and PLS method of chemometrics. It is applicable to forecast the MON, RON and PON of gasoline of catalytic cracking, catalytic reforming, alkylating as well as blended products. A near infrared optical multichannel analyzer (NIR-OMA) and a software package of octane number were developed. Combining both mentioned above a NIR-OM octane number analyzer was developed. With this NIR-OMONA one can obtain the MON, RON and PON (antiknock index) data in less than 1/2 minute with an accuracy of 0.6 ON unit and a precision of 0.1 ON unit by using 3-15 mL gasoline sample. PMID- 15822268 TI - [Study on the inkblot of carbon pen fields by Raman micro-spectroscopy]. AB - The four carbon-pen inkblots in market were studied by Raman micro-spectroscopy. The Raman spectra obtained in the experiments indicated that four brands of carbon-pen have not only similarity but also differentiation. Raman spectra from the same brand but different batch demonstrated difference too. Hence one can use the Raman technique for characterization and identification of inkblots from market. PMID- 15822267 TI - [Vibration studies of amphetamine and methamphetamine by micro Raman]. AB - For the first time, we obtained and investigated the Raman spectra of amphetamine and methamphetamine by micro Raman with incidence excitation light 514.5 nm under the ambient environment. According to the fact that both molecules belong to single substituted benzene which has the symmetry of Cy2v, the attribution of Raman vibration modes in amphetamine and methamphetamine molecules was carried out by studying the spectral lines in the two spectra. The Raman line that only appeared in methamphetamine but could not be seen in any other drugs we have investigated was most possibly attributed to the stretching vibration between C N(+)-C. PMID- 15822270 TI - [Analysis of the spectrum of solid sodium carboxyl in N+ ion implantation]. AB - The changes in molecular structure of PhCOONa and CH3COONa samples in 25 keV N+ ions implantation were measured by UV-spectrum. The dose response of UV absorption was deduced. It was found that a new compound with UV-absorption could be produced in implanted CH3COONa sample. The cyano-group could be produced in PhCOONa and CH3COONa samples implanted by the N+ ion beam.samples implanted by the N+ ion beam. PMID- 15822269 TI - [Determination of trace iron (II) and molybdenum (VI) in ground water by pyrocatechol resin-phase spectrophotometry]. AB - In weak acid medium (pH 6.20), Fe(III) and Mo(VI) can form colored complexes with pyrocatechol and quantitatively adsorbed on 717 strong base anion exchange resin. The apparent molar absorption coefficients of resin phase are epsilon = 4.1 x 10(4) L/(mol x cm) at 520 nm for Fe(III) and epsilon = 9.0 x 10(4) L/(mol x cm) at 400 nm for Mo(VI). Beer's law is obeyed for Fe (III) and Mo(VI) in the range of 0-2.2 mg x L(-1) and 0-1.6 mg x L(-1). The method has been applied to the determination of trace Fe(III) and Mo(VI) in ground water. The relative standard deviations are 3.3% and 3.2%, respectively. PMID- 15822271 TI - [Effect of the coordinate micro-environment on the structure of Hg (II)-serum albumin]. AB - Detailed studies on the UV spectra of Hg (II)-HSA or Hg (II)-BSA complexes nearby isoionic point were carried out in this paper. Compared with the spectra at physiological pH, the band nearby 250 nm of Hg (II)-BSA generally disappeared or widened and weakened,but the band at 290 nm of Hg (II)-BSA and Hg (II)-HSA remained nearly unchanged. These results support that these binding sites of Hg (II)-serum albumin may be located at seven pairs of adjacent disulfide bridges and there might be the structure of HgS4 tetrahedral configuration. For the first time, this paper put forward the solid figure of HgS4 tetrahedral configuration, and further discussed the results using the discrepancy of the coordinate micro environment. PMID- 15822273 TI - [Synthesis of methyl ester of aliphatic acid under irradiation of ultraviolet and mass spectra (MS) analysis]. AB - A new method was introduced here to prepare methyl ester of aliphatic acid under irradiation of ultraviolet. Analysis was performed for the MS spectra of 19 products, by which Mclafferty rearrangements were discussed of methyl ester of aliphatic acid with two kinds of gamma-H in details. The regularity of Mclafferty rearrangement was established for the same kind of methyl ester with the increase of the carbon chain. PMID- 15822272 TI - [Study of conformation of serum albumin by FTIR]. AB - This paper used the FTIR method to study the spectra of bovine serum albumin (BSA) and its solution. The secondary structure and conformation was assigned by FTIR-deconvolution analysis. The result indicated that BSA secondary structure in solid is different from in aqueous solution. As the decrease of BSA concentration from 3.241 x 10(-1) to 4.032 X 10(-2) g X mL(-1), the amide I band components at 1 609.84, 1 633.85,1 653.69, 1 681.16 and 1 694.88 cm(-1) shifted to 1 608. 24, 1 638.36, 1 656.10, 1 674.87 and 1 690.78 cm(-1), respectively, the component at 1 621.50 cm(-1) unchanged. PMID- 15822274 TI - [Software package for self-constructed CCD-ICP-OES. 1. Software system and design idealogy]. AB - A software package and its design idealogy for the self-constructed CCD-ICP-OES are described. The MS Windows 95TR was used as the operating platform. The Software package written in MS Visual C + + TR V 4.2 included the control of grating and CCD, the dynamic monitoring, the treatment of spectral signal, the fixing of wavelength position, the automatic fixing of spectral line, the quantitative analysis, the stripping method, the Kalman filter and the artificial neural network. PMID- 15822275 TI - [Determination of impurities in osmium powder by ICP-AES using matrix separation]. AB - An inductively coupled plasma-atomic emission spectrometry method has been developed for the determination of impurities in osmium powder. The sample was dissolved in nitric acid at 150-200 degrees C. Osmium was oxided and its oxide vapour was absorbed by NaOH. The residual solution was then analyzed by ICP-AES. The determination limit of the method is 0.005-0.134 microg x mL(-1). The recovery range of the method is 91%-112%. The precision of the method is less than 3%. This method can be used to determine 18 elements simultaneously. It is simple, rapid and precise. PMID- 15822276 TI - [Study on the emission intensities of metal samples in a laser microsource]. AB - The reflectance of laser beam on metal surface was considerably decreased by means of daubing a coating or changing the smooth finish. The emission intensity of metal samples was increased in a laser microsource. Therefore, the detection limit of spectrochemical analysis was reduced. PMID- 15822277 TI - [Determination of inorganic and organic tin in water samples by graphite furnace atomic absorption spectrometry]. AB - This paper proposed a method for the determination of inorganic tin and organotin species. Total tin contents can be directly detected by zirconium-coated graphite furnace atomic absorption spectrometry,while the organotin was detected by hydride generation combined with solvent extraction and the difference between the total tin and organotin was inorganic tin. The detection limits were 58 pg and 27 pg for organotin and inorganic tin,respectively. The relative standard deviations of this procedure, were 2.13% and 2.04%, respectively. Several water samples were analyzed by this method. The recoveries ranging from 90.5% and 98.5% were obtained. PMID- 15822278 TI - [Determination of trace amounts of zinc in nickel electrolyte by flow injection on-line enrichment]. AB - A method for the determination of trace amount of zinc in nickel electrolyte utilizing the flow injection on-line enrichment technique is reported in this paper. Atomic absorption spectrometer was used as detector. Zinc was separated from large amounts of nickel andother components in the electrolyte by absorption its chlorocomplex on a mini-column packed with strongly basic anion exchangers. It was found that sodium chloride containing in the electrolyte offered a sufficient chloride concentration needed for the formation of the zinc chlorocomplex and thus no additional reagent was required for the determination. The throughput of the method is 30 determinations per hour. The detection limit of the method is 0.002 microg x mL(-1) and the precision is 1.9% (RSD). The proposed method is rapid and cost-effective. It has been used for almost three years in the quality control of the electrolyte in the factory with great success. PMID- 15822279 TI - [Determination of trace barium in biological samples by Zeeman graphite AAS with coated graphite tube]. AB - Using a tungstate-coated graphite tube, trace barium in biological samples was determined by Zeeman graphite AAS. The sensitivity of Ba can be significantly improved. The precision and the lifetime of graphite tube have been improved by adding matrix modifier. The method is simple. The recovery and precision are satisfactory. PMID- 15822280 TI - [Interferences in analysis of mineral elements in salt by FAAS and their correction]. AB - This paper studies interference factors in the analysis of mineral elements in salt by FAAS and discusses the problems of background interference, Cl- interference, Na+ interference and other ion interferences. K, Na, Mg, Cu, Mn, Zn, Fe, Pb, Cr and Cd in commercial salt were determined. The results showed that this method did not suit Cr,background absorption only interfered with the determination of K, and Mg, 1% Cl- interfered with the determination of K, and 10% Cl- or 2% Na+ interfered with the other elements. After technical treatment and eliminating all sorts of interferences, the results of analysis for salt are satisfactory. PMID- 15822281 TI - [Quasi-liquid membrane enrichment separation and determination of trace magnesium and lead by flame atomic absorption spectrometry]. AB - This paper reports a new method of separation and enrichment of trace magnesium and lead quasi-liquid membrane. Under optimum conditions for the determination, the recovery is more than 90%. The method is simple, accurate and practical. PMID- 15822282 TI - [X-ray photoelectron spectroscopy (XPS) of carbon nitride (CN) films]. AB - The chemical structure of carbon nitride thin films, prepared by rf-dc plasma enhanced chemical vapor deposition (PECVD) was studied by XPS. Analyzing the C(1s) and N(1s) core level lines indicated two types of chemical structure N-sp3C and N-sp2C binding states existed in the CN films. There is also little N-spC binding state in the CN film with a nitrogen content of 22%. The ratio of N/C in N-sp3C binding state is 1.28 which is near 4 : 3, thus demonstrated the existence of C3N4-like phase. High nitrogen content in the CN films is useful to increase the content of beta-C3N4 phase. Light changes of the chemical structure of the CN films can be observed under ion irradiation. With the increase of ion dose, the ratio of N-sp3C/N-sp2C increase, while the N/C ratio in the films decrease. PMID- 15822283 TI - [Determination of Pb, Cu and Sn in ancient bronze coins using X-ray fluorescence theoretical alpha coefficient method]. AB - In this paper, the theoretical alpha coefficients of L-T equation were applied to the analysis of Pb, Cu and Sn in ancient bronze coins by X-ray fluorescence spectrometry. The results obtained in this way are in agreement with those by the fundamental parameter method and the simulant standard samples method, because this method and the simulant standard samples method were both developed from the fundamental parameter method, and the standard samples are the same. The calculation of element concentration is simple and rapid, but the precision of the results depends relatively largely on the concentration range of the standard samples by this method. PMID- 15822285 TI - [Sensitizing effect of nonionic microemulsion on the derivative spectrophotometric determination of trace iron with sulfosalicylic acid]. AB - The sensitizing effect of O/W microemulsion of Triton X-100/n-C5H11OH/n-C9H20/H2O as a medium on the derivative spectrophotometry of Fe-Sal was studied. The results show that the sensitivity of determination in O/W microemulsion was higher than that in micelle. The reagent Sal has higher distribution coefficient KD and constant K in microemulsion than in the micelle system and lower binding in O/W microemulsion than in the micelle system. This results also illustrate that O/W microemulsion has more solubilization for Sal than micelle does, so that O/W microemulsion has more sensitizing reaction than micelle does. PMID- 15822284 TI - [Monitoring and elimination of sulfur dioxide and nitrogen dioxide by passive sampling method]. AB - The concentration of atmospheric pollutants, sulfur dioxide and nitrogen dioxide, were monitored by passive sampler with spectrophotometric determination method. The results were compared with active sampling method. It was found that the two methods got the same results. The adsorption of sulfur dioxide and nitrogen dioxide by sorbent-saturated filter paper was also studied. PMID- 15822286 TI - [Dimerization of aluminum, zinc, vanadium and gallium phthalocyanine-sulfonates in water, aqueous alcoholic solutions and micelles]. AB - The dimerization of Aluminum (III),Vanadium(lV), Gallium(III) tetrasulfonated phthalocyanine (AlTSPc, VOTSPc, GaTSPc) and Zinc(II) bisulfonated phthalocyanine (ZnBSPc) was studied by spectrophotometic method. The dimerization constants KD were calculated from absorbance measurements. The absorption spectra in water, aqueous alcoholic solutions and micelles of phthalocyanine sulfonates aluminum shows the "slow-forming dimer", and VOTSPc, GaTSPc, ZnBSPc show contact dimer formation. PMID- 15822287 TI - [Synthesis of the new reagent N-hexyl-N'-(sodium aminobenzenesulfonate) thiourea (HXPT) and its spectrophotometric study with Pd2+]. AB - A new reagent N-Hexyl-N'-(sodium aminobenzenesulfonate) thiourea (HXPT) was synthesized. Its structure was also characterized. A new spectrophotometric method of determining Pd2+ was developed based on the new reagent. In the range of pH 5.4-5.8 Pd2+ forms a stable yellow complex with HXPT in the presence of CTMAB,and its apparent molar absorptivity is 2. 21 x 10(5) L x mol(-1) x cm(-1) at the maximum absorption wavelength 296.8 nm. Beer's law is obeyed in the range of 1-7 microg x 25 mL(-1) for Pd2+. This method has been used for the determination of trace Pd2+ in ores and catalyst with satisfactory results. PMID- 15822288 TI - [Determination of the content of aminopyrine in antongding injection by orthogonal function spectrophotometry]. AB - Determination of the content of aminopyrine in the antongding injection by orthogonal function spectrophotometry was reported in this paper. The six wavelengths for the determination are 257, 261, 265, 269, 273 and 277 nm. The concentration of aminopyrine in the range of 5-25 microg x mL(-1) is in a good linear relation to P2, and the regression equation is P2 = + 0.018 - 0.032 9c, r = 0.999 9. The average recovery and relative standard deviation are 100.3% and 0.40%, respectively. On the basis of the data observed it is concluded that this method shows no obvious difference as compared with Hebei province pharmacy regulation. PMID- 15822289 TI - [Measurement of aluminum (III) in crops by synergistic chromatic reaction]. AB - In the presence of OP, a sensitive color reaction of aluminum (III) on pyrocatechol violet and rhodamine 6G has been studied. In a NH4Ac-HAc (pH = 6.5) medium, aluminum (III) ions form an ions-associate complex with pyrocatechol violet and rhodamine 6G. The composition of the complex is Al (III) : PV: Rh6G = 1 : 2 : 4. The maximum absorption of the ions-associate complex is at 580 nm with an apparent molar absorptivity of 6.8 x 10(4) L x mol(-1) x cm(-1). Beer's law is obeyed in the range of 0-7 microg/25 mL of aluminum. The method is highly selective and has been applied to the determination of trace aluminum in crops with satisfactory results. PMID- 15822290 TI - [Study on the new method for spectrophotometric determination of potassium iodate in salt]. AB - A new simple, fast and sensitive spectrophotometric method in water phase for the determination of micro amount of potassium iodate in salt was studied. The method is based on chromogenic reaction of crystal violet on I3- produced from potassium iodate reacting on KI in HCl medium and using OP as the solubility-increasing reagent. When applying the proposed method to determining potassium iodate in salt, the results obtained were in agreement with those of ultrovilet spectrophotometry. PMID- 15822291 TI - [Study on the synergistic coloration of tin (II) with pyrocatechol violet and rhodamine B and its application]. AB - In the presence of PVA, a sensitive color reaction of tin (I ) on pyrocatechol violet and rhodamine B has been studied. In a NaAc-HAc (pH = 5.0) medium, tin(II) ions form an ions-associate complex with pyrocatechol violet and rhodamine B. The composition of the complex is Sn (II): PV : RhB = 1 : 2: 4. The maximum absorption of the ions-associate complex is at 560 nm with an apparent molar absorptivity of 1.18 x 10(5) L x mol(-1) x cm(-1). Beer's law is obeyed in the range of 0-15 microg/25 mL of tin. The method is highly selective and has been applied to the determination of trace tin in some objects with satisfactory results. PMID- 15822292 TI - [The analysis of pressure shifting and broadening coefficients from multiple-line photoacoustic spectra of water vapor]. AB - The pressure shifting and broadening coefficients of water vapor were obtained from the photoacoustic spectra with multiple lines. The Voigt spectral profile and parameter confined optimizing by group were adopted to obtain high precision and efficiency. PMID- 15822293 TI - [Spectral studies on the mechanism of rare earth poly (vinyl chloride) thermal stabilizers]. AB - Rare earth Poly (Vinyl Chloride) (PVC) thermal stabilizers are one kind of effective and non-toxic stabilizers developed recently. By photoacoustic (PA) spectra and FT-IR analysis, their excellent performance in PVC stabilization progress can be studied conveniently. Good color remaining ability of rare earth stabilizers can be shown clearly by PA spectra and compared with others easily. From the IR spectrum and process data, the extra stabilization ability of rare earth stabilizers may be attributed to their conformation transforming effect. PMID- 15822294 TI - [Study on luminescence of the serial new ternary or tetradic complexes complexes of Eu (III) with beta-diketone]. AB - The UV absorption, fluorescence, unstable spectra and quantum yields of five ternary or tetradic complexes of Eu (III) with beta-diketone (dibenzoyl methane, thenoyltrifluoroacetone), acrylic acid and other ligands were reported. The influences of the first and second ligands on absorption and luminescence spectra were found. The lifetimes and the processes of energy transfer of Eu3+ from 5D1 to 5D0 in complex solutions were measured. It was found that the lifetimes of Eu3+ complexes with TTA are approximate to each other and larger than those of complexes with DBM. PMID- 15822296 TI - [Synthesis and ion recognition function of 7-methoxy-4-methene-coumarin-monoaza 18-crown-6 (I) synthesis and spectroscopic properties]. AB - The title reagent MMC-MAC (O5) is synthesized and characterized. Its acidic dissociation constant at ground and excited state are determined based on the changes of the absorption and fluorescence spectra in the water. According to the wavelength shift of fluorescence spectra in different solvent, the dipole moments of ground and excited state are estimated based on the molecular mechanics model using a alchemy 2000 software and the solvachromic method using the Lippert equation,the value is 3.11D and 8.13D,respectively. It's also found that fluorescence intensity of MMC-MAC(O5) aqueous solution is increased marvelously about twenty times and accompanying with delay fluorescence emit when deoxygenation from the solution by H2SO3-N+ (CH3) 4OH-. The intensity of normal fluorescence and delay fluorescence are proportional to the concentration of MMC MAC(O5) in the range of 4 x 10(-8)-2 x 10(-6) and 2 x 10(-7)-2 x 10(-6) mol x L( 1), and the detection limits are 2 x 10(-9) and 1.3 x 10(-8) mol x L(-1) respectively. PMID- 15822295 TI - [Changes of infrared spectra for amino acids induced by low energy ions]. AB - Low energy ions have been generated from implanter and gas arc discharge at normal pressure, and impacted on amino acids in solid state and in aqueous solution. The induced changes of infrared spectra have been investigated. It showed that ions generated by these two means have the same or similar damage effects, such as rearrangement of damaged molecules and deposition of external ions, and the damage effects are especially remarkable and various when ions attack molecules in solution. PMID- 15822297 TI - [Effect of electric field on optical radiation linewidth of free-carriers in quantum wells]. AB - By using concepts as orbit, momentum and wave packet, the particles which move in a large range of space in solid state are described. According to the uncertainty relations in quantum mechanics, a formula of energy uncertainty of particles is derived. The formula indicates that electric field scatters carriers strongly and makes energy levels broaden largely. Applying the formula to GaAs/GaAlAs quantum wells with P-I-N structure, the calculated optical radiation linewidths are in agreement with measured photoluminescence linewidths. PMID- 15822298 TI - [Study on the interaction of human serum albumin and Mn (II), Co (II) by fluorescence method]. AB - The interaction of Mn (II), Co(II) and HSA has been studied by fluorescence method at pH 7.4 and 5.3. Based on Forste non-radiative energy transfer theory, the distance between Trp-214 residue of HSA and the first strong binding site to Mn (II), Co (II) in HSA was determined, and it is much larger than that of reference. This notable different result has been discussed according to the binding site of Mn (II), Co (II ) and domain structure in HSA. PMID- 15822299 TI - [Anharmonicity of local vibrational mode of carbon acceptor in GaAs and its effect on main absorption band]. AB - A sideband on the low energy side of the local-vibrational-mode main absorption band of carbon acceptor in GaAs was observed. The difference between the peak frequency of the main absorption band and that of the sideband is 3.5 cm(-1). The integrated absorption of the sideband varies with temperature and has an effect on the temperature dependence of the main absorption band. It is likely that the sideband originates from the anharmonicity of the local vibrational mode itself. PMID- 15822300 TI - [Comparative studies on the absorption properties of charge transfer complexes of C60 with three organic amines]. AB - Comparative studies on the absorption properties of charge transfer complexes of C60 with N,N-dimethylaniline, diethylamine and triethylamine in toluene, CCl4 and cyclohexane were carried out, respectively. It was found that interactions of C60 with various amines in different solvent media are all in the order: cyclohexane > CCl4 > toluene, while these interactions are different in the same solvent. Both of the molar extinction coefficients and apparent constants of CTC were determined. Meanwhile, the influences of temperature on CT interactions of C60 with various amines in different solvent media were also studied. PMID- 15822302 TI - [Study on the interaction of copper-zinc superoxide dismutase with cobalt (II) histidine by spectral analysis. II. Effects of pH, interaction time]. AB - The direct interaction of Cu2Zn2SOD with organic metal compound (Cobalt (II) Histidine) was studied by ICP, VIS and the measurement of enzyme activity, and also investigated the effect of pH values and different interaction times for this interaction. The results showed that with the increased in pH values and the more longer interaction times, the intensity of interaction were increased and the corresponding catalytic activity of enzyme were affected. PMID- 15822301 TI - [Study on the interaction of copper-zinc superoxide dismutase with cobalt (II) histidine by spectral analysis--II. Effect of amount of external added Co(His)n, phosphate]. AB - The effect of changing the amount of external added Co(His)n and phosphate on the interaction of Cu2Zn2SOD with organic metal compound (Cobalt (II)-Histidine) have been studied by means of ICP, VIS and measurement of enzyme activity. It has found that in aqueous solution, there exists a direct interaction of the metal ions of the active center in the metalloenzyme (Cu2Zn2SOD) with external added Co(His)n. As a result, part of the metal ions in metalloenzyme were replaced and the corresponding metalloenzyme derivatives (Co (II)-substituted derivatives of SOD) were produced and the catalytic activity of enzyme were affected. It has also studied the intensity of interactions in different molar ratios (Cobalt (II) Histidine) and of the presence of phosphate and got some results under the effect of corresponding factors. PMID- 15822303 TI - [Mechanism of gold solvent extraction from alkaline cyanide solution by surfactant 1 427]. AB - The solvent extraction of gold and the structure of the organic phase were studied by labelled 198Au, 31P NMR and FTIR spectroscopy. The results show that the water content increased with the increase of gold contents in the organic phase. The P=O stretching decreases, while the 31P signal shifts to upfield. PMID- 15822304 TI - [Surface-enhanced Raman spectroscopy of riboflavin in silver colloids using 1 064 nm excitation]. AB - Surface-enhanced Raman spectra (SERS) of riboflavin in silver colloids, using near-infrared (NIR) laser operating at 1 064 nm wavelength, were obtained. The enhancement factor was further increased by improving the preparation of silver colloid and with the addition of a small amount of NaOH. The detection limit was lower than 3.2 x 10(-10) mol x L(-1). It is concluded that the larger and aggregated silver particles were more efficient for the electromagnetic enhancement of NIR-SERS. Flavin ring was absorbed on the colloidal silver methal surface, standing on oxygens of C(20)=O) and C(4)=O. PMID- 15822305 TI - [Raman spectra of complexes of rare earth nitrate with histidine]. AB - Raman spectra of solid complexes RE(His)(NO3)3 x H2O (RE = La-Nd, Sm-Lu, Y; His = L-alpha-histidine ) have been investigated. The results indicate that RE3+ coordinates with one O atome of carboxyl group in the complex, while amino group and imidazole ring do not take part in coordination and NO3 is double coordination. The vibration peaks of carboxyl group delta(v)COO-(as-s) were plotted against the atomic number of the lanthanoids, which obeys Oddo-Harkins law. PMID- 15822306 TI - [Study on chitosan-Zn (II) complex by IR spectroscopy]. AB - A series of chitosan-Zn (II) complexes were synthesized by reaction of chitosan and ZnSO4 in different ratios(chitosan : Zn (II) = 1 : 0.05-1 : 1) in dilute formic acid aqueous solution. The results showed that -NH2, -OH and -NHCO- of chitosan molecule were coordinated to Zn (II) based on the analyse of the IR spectra of chitosan, chitosan-H2SO4 and chitosan-ZnSO4. SO4(2-) of chitosan-ZnSO4 may be crosslinking of chitosan molecule, but Cl-, CH3COO- and NO3- were not crosslinked to chitosan molecule in the chitosan-ZnCl2, chitosan-Zn (CH3COO)2 and chitosan-Zn (NO3)2 complexes. PMID- 15822307 TI - [Synthesis and characterization of complex La(C5O3H3)3 x C12H8N2 x H2O]. AB - The title ternary complex La(C5O3H3)3 x C12H8N2 x H2O was synthesized by reaction of lanthanum nitrate with alpha-furoic acid (C5O3H4) and 1,10-phenanthroline (C12H8N2) as ligand in the alcohol solvent. The composition of the complex was determined by element analysis. Its structure and properties were characterized by IR, 1H NMR, UV and TG-DTA. IR, 1H NMR, UV spectra showed that the ligand alpha furoic acid coordinate to the center ion La (III) with the deprotonized form, and 1, 10-10phenanthroline with the bidentate form using its two N atoms. The TG-DTA analysis exhibited that the complex is stable in the range of room temperature and undergo the disassociation of water and oxygenized decomposition of the organic ligands in order with the rising of temperature, and the final oxide obtained is La2O3. The quantitative analysis of TG-DTA was coincided with the structural composition of the title complex. PMID- 15822308 TI - [Study on the ingredients of reserpine by TLC-FT-SERS]. AB - A new method for analysing the ingredients of reserpine by thin layer chromatography (TLC) and surface-enhanced Raman spectroscopy (SERS) is reported in this paper. The results show that the characteristic spectral bands of reserpine satuated at the thin layer with the amount of sample about 2 microg were obtained. The difference between SERS and solid spectra was found. An absorption model of reserpine and silver sol was proposed. This method can be used to analyse the chemical ingredients with high sensitivity. PMID- 15822309 TI - [Studies on the infrared spectra of rare earth complexes with pyruvic acid]. AB - FT-IR spectra of La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho and Y complex with pyruvic acid have been studied. The principal infrared absorption peaks are assigned for the region 400- 2 000 cm(-1). Three IR peaks in the region 200-400 cm(-1) are shifted obviously to higher frequency with increasing of the atomic number of rare earth ion, these were believed to be RE-O bond stretching mode absorptions. The IR spectra also indicated that the carboxylate groups have the form of chelation bridging tridentate to rare earth ions, i. e., one of the carboxyl oxygens was shared between two adjacent metal ions, moreover, the splitting and broad bands are existed in IR spectra of the rare earth complexes. The RE-O bond of the complexes is principally ionic. PMID- 15822310 TI - [Monitoring glucoamylase fermentation with infrared spectroscopy]. AB - The glucoamylase fermentation was investigated with Fourier transform infrared spectroscopy. During the cultivation, the variation of carbon and nitrogen source can be reflected from the infrared spectra. In most cases, the complex media are used to cultivate cells or bacteria. It is difficult to construct calibration samples with synthetic mixtures which can represent the composition of real broth. The infrared spectral data of the real broth and off-line chemical analysis results were used to establish the calibration model. The model is successfully used to predict the concentrations of total sugar, reduced sugar, and nitrogen in real broth. The correlation coefficients between the infrared analysis results and chemical analysis are 0.999, 0.990 and 0.974 for total sugar, reduced sugar and nitrogen, respectively. The bacteria, culture and operating condition are relatively stable during the practical productive processes. The method can be used to monitor the industrial processes. PMID- 15822311 TI - [Analysis of blue ballpoint ink components by FT-IR microspectrometry]. AB - Systematical analysis on the 108 kinds of blue ballpoint writing inks with FT-IR microspectrometry is presented in this paper. The results show that the differences of the ink compounds keep some regular. These blue ballpoint writing inks can be classified by the transmittance spectra. The method is rapid, accurate, highly sensitive and nondestructive. PMID- 15822312 TI - [Studies of narcotics by micro Raman spectroscopy]. AB - For the first time, the Raman spectra of eight kinds of narcotics were obtained and investigated by micro Raman. The results indicated that the Raman spectra were affected by the numbers and position of methyl in substituent groups. Under incidence light 514.5 nm and appropriate measurement condition, high quality Raman spectra of drugs with low fluorescence background were obtained. In addition, we discovered that narcotine and thedaine were very sensitive to strong light radiation. Finally, by comparing Raman spectra carefully for all drugs we measured the specific Raman peaks were given for the purpose of distinguishing different kinds of drugs. PMID- 15822313 TI - [New researches in the direct determination of Chinese traditional drugs with molecular spectroscopy]. AB - The article relates to the latest development of NIR FT-Raman and DR-FTIR technique and their special use in the determination of Chinese traditional drugs. PMID- 15822314 TI - [Artificial Neural Network and its application to analytical chemistry]. AB - Artificial Neural Network system(ANNs) was presented involving its theoretical basis, algorithm and performance. Its application to nonlinear calibration, pattern recognition and other related fields in analytical chemistry has been reviewed. 95 references are cited. PMID- 15822315 TI - [Study on magnesium ionization in cathodic sputtering glow discharge plasma]. AB - A method, based on the Doppler broadening-dependent absorption width and the ionization degree measured by the ratio of relative atomic and ionic absorbance in cathodic sputtering glow discharge (CSGD) plasma with aluminum-magnesium alloys as cathodes, is proposed. The experimental reveals that the plentiful magnesium ions in the plasma under the conventional discharge conditions of the atomizer do influence the analytic working curves. Corrected method is derived and factors which result in magnesium ionization degree are disscused. PMID- 15822316 TI - [Determination of the impurities elements in high-purity cadmium by inductively coupled plasma mass spectrometry]. AB - A method for the determination of impurity elements in high purity cadmium by inductively coupled plasma mass spectrometry was developed. The spectral interference arising from Cd was discussed and the magnitude of interference by the polyatomic ion of Cd was compared. The effect of the high concentration Cd on the analyte signal was studied. The results showed that Cd possessed both significant suppression effects on the light mass elements signal and enhancement effects on the heavy mass elements signal, and the effects can be corrected by the use of internal standard elements. In this paper, Y and Bi was employed for the light mass elements and the heavy mass elements, respectively. As, Be, Co, Cu, Ga, Ge, Mn, Mo, Pb, Ni, Sr, Au, Tl, Th, V and U in high purity cadmium were determined. Detection limits are 0.005-0.052 microg x L(-), and the recoveries of standard addition are 82%-108%. PMID- 15822317 TI - [Software package for self-constructed CCD-ICP-OES. 2. Algorithm bases for the softwares]. AB - The algorithm bases for the Savitzy and Golay smoothing, spectrum stripping, Kalman filtering and the artificial neural network programs used in the self constructed CCD-ICP-OES are described. PMID- 15822318 TI - [Determination of cadmium in soil slurry samples by GFAAS using ammonium oxalate as disperstant]. AB - A method for the determination of trace cadmium in soil slurry samples by GFAAS was optimized. The factors that influence the performances of cadmium at different chemical modifier were investigated. The effect of background has also been studied. It is found that the slurry of soil samples was most stable with ammonium oxalate as disperstant. The results show that the determination of trace cadmium in soil slurry samples could performed. PMID- 15822319 TI - [Determination of trace Se in water by AA-670 atomic absorption spectrophotometry attached VA-90 atomization]. AB - This paper reports the determination of trace metal Se in water by AA-670 atomic absorption spectrophotometry attached VA-90 atomization. The method is convenient to determine element in water. The results show that the method has good precision and accuracy. The relative standard deviation of Se was 2.7%, and the rates of recovery were 94.2%-101.6%. PMID- 15822320 TI - [Analysis of 4 metal elements in 11 kidney tonic Chinese traditional medicines]. AB - The contents of Zn, Cd, K and Na in 11 kidney tonic Chinese traditional medicines usually used in clinical practice were determined by FAAS. The results showed that element Zn, which is closely related to yang deficiency of the kidney, and the macro elements K and Na, which are essential to human-body, are very rich, but element Cd is lower. The relations between trace elements and the therapeutic efficacy were discussed, and it is concluded that the results are in agreement with clinical usage of medicines by herbalists. PMID- 15822321 TI - [Determination of mineral elements in milk and milk products extracted with hydrochloric acid(HCl) by FAAS]. AB - Mineral elements K, Na, Ca, Mg, Fe and Zn in milk and milk products were extracted with boiling 1.0 mol x L(-1) hydrochloric acid and determined with atomic absorption spectrometry (AAS). This procedure proved simple, safe and quick. The results obtained, either for liquid samples or for solid ones, were accurate, precise and comparable with those by dry ashing. PMID- 15822322 TI - [Quantitative analysis of Pt-Pd alloys using partition polynomial regression]. AB - A new regression method was developed for X-ray microflurescence analysis of Pt Pd alloys using X-ray tube excitation. Working curve is divided into four parts based on concentration of Pt in alloys. For each part, polynomial regression method is used to fit the curve. Results show that this procedure is superior to conventional method of polynomial regression which fitting the whole curve with one equation. Homogeneous level of element distribution in standard alloys is also investigated PMID- 15822323 TI - [Transmission grating spectrograph for soft X-ray spectrum measurements with a pre-optics]. AB - A novel diffraction spectroscopic instrument consisting of a large area transmission grating and a grazing-incidence pre-optics comprising of a toroidal mirror, a slit and a cylindrical mirror has been implemented. A nearly perfect stigmatism and a medium spectral resolution are due to the separation of the focusing and dispersing function. The experimental results show that it is possible to construct large area transmission grating spectrograph with high collecting efficiency and good spatial resolution. PMID- 15822324 TI - [Determination of sulfur dioxide by fluorometric analysis]. AB - A method to determine the concentration of atmospheric pollutant, sulfur dioxide, was studied by fluorophotometric method. The composition and concentration of reaction solution were optimized. This method can be developed to be a fibre optic fluorometric method for routine monitoring. PMID- 15822325 TI - [Study on properties of fluorescence and room temperature phosphorescence of 2 bromomethyl naphthalene]. AB - The properties of fluorescence and room temperature phosphorescense (RTP) of 2 Bromomethyl naphthalene (2-BrMN)were studied. The results show that the wavelength for maximum fluorescence of 2-BrMN is lambda(ex/lambda(em) = 274/334 nm. The fluorescence intersity is proportional to the concentration of 2BrMN in the range of 1.0 x 10(-6) - 1.2 x 10(-4) mol x L(-1), and the detection limit is 4.7 x 10(-8) mol x L(-1). The RTP signal of 2-BrMN was induced by beta cyclodextrin (beta-(CD) in the presence of dibromopropane (DBP) acting as a heavy atom perturbation. The RTP intensity is linear for 2-BrMN/beta-CD/DBP systems of 2-BrMN concentration in the range of 1.0 x 10(-5) - 1.2 x 10(-4) mol x L(-1) (lambda(ex)/lambda(em) = 271/493,521 nm), and the detection limit is 2.7 x 10(-6) mol x L(-1). PMID- 15822326 TI - [The hydrogen bond between 5-(2-carboxyphenyl)-10, 15, 20-triphenylporphyrin and amino acid ester--study with UV-Vis titration and 1H-NMR]. AB - The hydrogen bonds between the hydrogen of pyrrols in 5-(2-carboxyphenyl) 10,15,20-triphenylporphyrin and the oxygen of the carboxyl group of amino acid ester were confirmed by UV-Vis spectroscopic titration method and 1H-NMR. When amino acid ester was added to o-TPPCO2H solution in CHCl3, the Soret and Q and red-shifted dramatically and several isobestic points could be observed in UV-Vis spectra. In 1H-NMR, the signal of the hydrogen atoms of pyrrole in o-TPPCO2H ( 2.78 ppm) decreased and a new signal in 0.260 ppm emerged, when o-TPPCO2H mixed with L-PheOEt at 1 : 1 (mol/mol). PMID- 15822327 TI - [Spectrophotometric determination of cinnarizine based on charge-transfer reaction]. AB - The charge-transfer (CT) complex formed between cinnarizine as the donor and 7, 7, 8, 8-Tetracyanoquinodimethane (TCNQ) as the acceptor in acetone-methanol has been studied by spectrophotometric method. Beer's law is obeyed in the range of 0 18 microg x mL(-1) of cinnarizine. The apparent molar absorptivity of CT complex at 743 nm is 1.58 x 10(4) L x mol(-1) x cm(-1). The composition of CT complex is found to be 1 : 1 by Bent-French and Job's methods. The relative standard deviation is less than 3% (n = 10). The method has been applied to the determination of cinnarizine in tablets with satisfactory results. PMID- 15822328 TI - Platyspondyly in childhood. PMID- 15822329 TI - The radiographic features of meningiomas in childhood and adolescence. PMID- 15822330 TI - The radiological findings in respiratory syncytial virus infection in children. Part I. Definitions and interobserver variation in the assessment of abnormalities on the chest X-ray. PMID- 15822331 TI - The systematic evaluation of the chest radiograph in cystic fibrosis. PMID- 15822332 TI - Four cases of lactobezoar in neonates. PMID- 15822333 TI - Ectopic ureterocele. A radiological appraisal of 66 consecutive cases. PMID- 15822334 TI - Hydronephrosis secondary to acute appendicitis in children. PMID- 15822335 TI - Bone disease in infants with prolonged obstructive jaundice. PMID- 15822336 TI - Pulmonary varix diagnosed in a newborn. PMID- 15822337 TI - Cholesterol ester storage disease. Radiological features. PMID- 15822339 TI - [Concept of cerebral mechanisms underlying prolonged adaptation to environmental hazards, according to I.M. Sechenov's scientific heritage]. AB - Occupational medicine is based on scientific ideas of Ivan Mikhailovitch Sechenov, the first Russian researcher of bioelectric activity of brain. Contemporary neurophysiologic methods of formalized quantitative evaluation of homeostasis in higher brain structures enable to optimize early etiologic diagnosis and prevention of central nervous and cardiovascular systems stress under exposure to multiple occupational hazards. PMID- 15822338 TI - Demonstration of vestibular implantation of ectopic ureters on an excretory urogram. PMID- 15822340 TI - [Prospects in getting accordance between chemical analytic control means and medical technical requirements to safety system concerning chemical weapons destruction]. AB - The major unit monitoring chemical weapons destruction objects is a system of chemical analyticcontrol over the technologic process procedures and possibility of environment and workplace pollution withtoxicchemicals and their destruction products. At the same time, physical and chemical control means meet sanitary and hygienic requirements incompletely. To provide efficient control, internationally recognized approaches should be adapted to features of Russian system monitoring pollution of chemical weapons destruction objects with toxic chemicals. PMID- 15822341 TI - [Methodic approaches using normobaric interval hypoxia and impulse magnetic field to increase nonspecific and anti-radio resistance in workers of ecologically unfavorable industries]. AB - Controlled normobaric interval hypoxytherapy and impulse low-intensity magnetic field application were proved to be the most promising methods that increase nonspecific resistance. Technical means to use those methods are specified. PMID- 15822342 TI - [Principles and criteria of occupational medicine standard in prevention of occupational deafness]. AB - The article represents materials on prevention of occupational deafness--one of the most prevalent occupational entities. PMID- 15822343 TI - [Hygienic forecasting and toxicologic evaluation of potential danger caused by new sulphenamide vulcanization accelerators]. AB - The article covers forecasting work conditions in oncoming production of sulphenamides T and DC--vulcanization accelerators, specifies leading occupational hazards, justifies values of MAC for sulphenamide T and approximate safe levels of action for sulphenamide DC in air of workplace. PMID- 15822344 TI - [Biochemical parameters in evaluation of liver functional state in workers engaged into general mechanical rubber goods production]. AB - Findings are that lipid peroxidation and antioxidant system are the most sensitive processes--that was proved by malonic dialdehyde accumulation, higher ceruloplasmin and lower catalase levels. PMID- 15822345 TI - [Analysing results of rehabilitation of patients with occupational diseases in St Petersburg]. AB - The authors studied rehabilitation of patients with occupational diseases. Rehabilitation of such patients via treatment courses with additional sick payment appeared ineffective. Most cases of occupational diseases are persistent. Even without disability group assigned, the patients remain with reduced occupational abilities for a long time. PMID- 15822346 TI - [Immunologic reactivity of patients with occupational allergic dermatoses]. AB - The article presents results of examination covering chemical metallurgic plant workers: 41% of the examinees have allergic skin diseases. Findings are that patients with occupational allergic dermatoses have variable degrees of T immunity suppression. PMID- 15822347 TI - [Severe lead poisoning case in accumulator plant worker]. AB - The authors describe a case of severe lead intoxication in tinman of accumulator batteries, who is under occupational exposure to high lead concentrations (up to 1.5 mg/cu m). PMID- 15822348 TI - [System automatically processing psychophysiologic parameters of pilots]. AB - For ergonomic evaluation of aircraft, the authors developed and subjected to flight testing a system automatically processing psychophysiologic parameters of pilots, providing post-flight automated PC processing of values registered during flight--cardiac rhythm, skin and sub-clothing temperatures, pulmonary ventilation and working load. PMID- 15822349 TI - [Renal clearance of nitrates after single injection of sodium nitrate to white rats]. PMID- 15822350 TI - [All-Russian scientific conference "Problems of environmentally induced health disorders in population of industrialized towns of South Ural with advanced ferrous metallurgy]. PMID- 15822351 TI - Health care is a necessity, not a right. PMID- 15822352 TI - Importation of prescription medications--a pharmacist perspective. PMID- 15822353 TI - Patient safety comes first. PMID- 15822354 TI - New program offers Missouri physicians free information technology assistance. PMID- 15822355 TI - New survey finds Missouri physicians poised to use technology. Electronic systems can cut costs, boost quality of care. PMID- 15822356 TI - Professional divorce: breaking up is hard to do. PMID- 15822357 TI - Using stem cells destroys life. PMID- 15822358 TI - Current status and issues of concern in diagnostic radiology. AB - I have tried to give a little history, discuss some current topics: PACS, inappropriate image utilization, CT screening, Night Hawk, molecular imaging, sonoscope, CAD, and LBBH. I hope these issues are more familiar to you now. The articles that follow from the faculty of the University of Missouri-Kansas City School of Medicine and Saint Luke's Hospital will bring you up to date regarding the current status of CT, MRI, ultrasound, nuclear medicine, mammography, and interventional radiology. PMID- 15822359 TI - Update of computed tomography and its increasing clinical applications. AB - With the introduction of Multi-Detector Row Computed Tomography we now have the ability to perform true high resolution isotropic imaging. The advantages and uses of Multi-Detector row Computed Tomography will be discussed as it affects the areas of cardio-vascular, pulmonary, renal, and gastro-intestinal radiology. PMID- 15822360 TI - Review and update of MRI for clinicians. AB - The goal of this review is to provide clinicians with basic information about MRI and its current available uses. Introductory information and updates about MRI for a broad range of topics are presented: mechanics of MRI, selection of MRI for diagnosis; popular uses of MRI (MRA, acute stroke, acute spinal compression fractures, cardiac MR, breast MR); MR spectroscopy, functional MRI, changes in MRI technology; and MRI safety. PMID- 15822361 TI - Everything's up-to-date in ultrasound. AB - There is a sense that ultrasound (US) is falling behind other technologies, and has taken a diminished role in modern radiology. Fortunately, there are newer innovations in ultrasound that seem very promising. 3-D ultrasound has aided in better imaging of multiple regions, including the uterus, breast, carotids, and even superficial musculoskeletal structures. Other recent innovations include the use of ultrasound contrast agents, for improved visualization, and utilizing high ultrasound energy to initiate chemo-embolization and clot thrombolysis. PMID- 15822362 TI - Update of nuclear medicine--PET imaging. AB - A new era in nuclear medicine imaging emerged with the recent clinical acceptance of Positron Emission Tomography (PET) scanning. By evaluating the metabolic activity of certain lesions, numerous cancers can now be detected and staged with high sensitivity and specificity. With the further advancements of PET/CT fusion, functional and anatomic detail of lesions can be acquired simultaneously with improved accuracy. PMID- 15822363 TI - Current topics of interest in interventional radiology. AB - Interventional Radiology utilizes imaging guidance (primarily fluoroscopy, computed tomography and ultrasound) to perform diagnostic and therapeutic procedures in a minimally invasive manner. This update highlights several current and newer interventional radiology options for treatment of uterine fibroids, interventional oncology procedures for liver tumors and metastatic disease, varicose vein treatment, carotid stenting, cerebral aneurysm coiling, and removable inferior vena cava (IVC) filters. PMID- 15822364 TI - New techniques for mammography screening: advantages and limitations. AB - Screening mammography is the primary method used for the detection of breast cancer. Computer assisted detection (CAD) systems, magnetic resonance imaging of the breast, and breast lymphoscintigraphy with sentinel lymph node biopsy are three new imaging techniques developed to assist in the detection and evaluation of breast cancer and will be described and discussed. PMID- 15822365 TI - Identification of the schizophrenia prodrome in a hospital-based patient population. AB - The identification of the schizophrenia prodrome remains a clinical challenge. We analyzed the clinical course of the prodromal symptoms of 24 patients with first time diagnosis of schizophrenia over a two year period. Their progress was tracked over three years. Patients in the study group reported to mental health care providers years before the schizophrenia diagnosis with subtle, but consistent, alterations in perception and behavior. Furthermore, they received an axis I diagnosis and were prescribed medications, which suggest that they constitute a distinct clinical population. PMID- 15822366 TI - Making difficult choices at the end of life: a personal challenge for all participants. AB - With the modern ability to sustain life well beyond its natural limits questions about restricting treatment, identifying patient preferences, and the expectation that psychological as well as physical needs be addressed are part of the challenge. Physician beliefs and values should be considered when treatment goals are discussed with patients nearing the end of life. This allows the physician to balance them with those of the patient and to place them in the context of professional obligations even when beliefs conflict. Sharing beliefs with patients may strengthen the relationships and minimize conflict, but this should be done cautiously and in context of the patients' needs and the clinical circumstances. Care should therefore be taken to respect the moral sensitivities of all participants when difficult choices are to be made at the end of life. PMID- 15822367 TI - Pharmacist-led DM delivers clinical, financial dividends. PMID- 15822368 TI - Taking a tougher stance on lifestyle factors. AB - With health care costs continuing to rise at double-digit rates, many payers have decided to hold participants responsible for unhealthy behaviors by actually hiking their health care premiums. Proponents of the approach maintain that it gets peoples' attention in ways that most other strategies have utterly failed. However, opponents argue that such penalties risk an employee/participant backlash that could cause more harm than good. Experts weigh in on both sides of the debate. PMID- 15822369 TI - Higher profile for migraine management spurs boom in DM offerings. AB - With more employers taking an interest in DM, the economic case has been broadened to include such factors as absenteeism and worker productivity. This trend has prompted many organizations to take a new look at managing migraines. Surveys suggest these debilitating headaches arc responsible for millions in lost productivity, and yet there are several effective treatment strategies. Further, developers believe even modest migraine DM programs can deliver an ROI. PMID- 15822370 TI - A tool for more accurate risk-prediction in acute coronary syndrome. AB - Standardized guidelines are fine, but one size does not fit all when it comes to cardiac patients. In fact, clinicians often struggle with deciding how aggressively to treat a patient who has suffered a heart attack or another form of acute coronary syndrome. However, a new risk-prediction tool can help guide clinicians to more precise conclusions regarding patient care. PMID- 15822371 TI - Research at Texas dental schools. PMID- 15822373 TI - Clinical evaluation of an orthodontic self-etching primer. PMID- 15822374 TI - Frictional evaluation of silica-coated ceramic brackets. PMID- 15822375 TI - Herpes virus infections: a review for the dental practitioner. PMID- 15822379 TI - The need for disability income insurance. PMID- 15822380 TI - Infection control for dental radiographic procedures. AB - The employment of proper infection control procedures in the dental office is necessary to prevent the spread of infectious agents. The potential for cross contamination between dental personnel and the patient is considerable when exposing intraoral radiographs. The dental office darkroom or daylight loader can also be a potential contamination source if good infection control measures are not followed. By utilizing PPE, plastic barriers and disinfectants, DHCPs can minimize the likelihood of problems with contamination. PMID- 15822381 TI - Oral and maxillofacial pathology case of the month. Lingual mandibular sequestration with ulceration. PMID- 15822382 TI - Improving your internal collections policy. PMID- 15822383 TI - [Urological problems encountered during pregnancy]. AB - The many anatomical and physiological changes occurring during pregnancy affect the entire urinary tract. These changes can induce various urological diseases, alter renalfunction and can be life-threatening to the foetus and the mother Some changes can persist post partum. Asymptomatic bacteriuria must be investigated and treated to prevent about 40% of pyelonephritis and a twofold higher premature delivery rate. Duplex ultrasound is the reference examination to diagnose renal colic due to stones and a low-dose IVU can be proposed whenever there is a doubt about the diagnosis. Treatment is conservative. Lower urinary tract symptoms are very frequent and some disorders, such as stress incontinence, can persist after delivery. This review article describes the anatomical and physiological changes observed during pregnancy and their pathological consequences on the urinary tract in order to detect, prevent and treat these disorders. PMID- 15822384 TI - [Ageing men's sexual functions decline and the erectile dysfunction (ED) increase]. AB - Association between ageing men's progressive falling of circulating androgen levels and ED is not clearly demonstrated. The analysis of all what have been written about this subject clearly proves that an androgenomodulation of erectile function exists. Indeed, the androgens seem to have an action on penile tissue innervation, on the structure and function of penile trabecular smooth muscle, on the penile endothelial function, as well as on the fibroelastic properties of the penile corpus cavernous. The addition of testosterone improves a great number of androgen deficiency in the aging male (ADAM). Recent studies demonstrated that all hypogonadal patients cannot successfully benefit of phosphodiesterase type 5 (PDE5) inhibitors. With these patients, the prescription of testosterone replacement therapy may improve the response of PDE5 inhibitors. PMID- 15822385 TI - [Retrospective study of laparoscopic retroperitoneal radical nephrectomy]. AB - OBJECTIVE: To analyse the results of laparoscopic retroperitoneal radical nephrectomy in terms of feasibility, morbidity and disease control. PATIENTS AND METHODS: Between October 1997 and May 2004, 42 laparoscopic retroperitoneal radical nephrectomies were performed in 33 men and 9 women for renal tumour. Three or four trocars were used depending on the operative findings in order to achieve oncological imperatives: primary control of the renal pedicle before mobilization of the kidney (the artery was clipped and the vein was sectioned by Endo-GIA), passage in the plane of radical nephrectomy and absence of direct contact with the tumour The operative specimen was extracted whole in an impermeable endoscopic bag by enlarging a trocar orifice to avoid any parietal contact. RESULTS: 21 right radical nephrectomies and 21 left radical nephrectomies were performed in patients with a mean age of 61.8 years (range: 36 to 83 years). The mean size of the kidney was 120 mm (range: 60 to 170 mm), for a mean weight of 430 g (range: 55 to 870 g). The mean tumour size was 46.4 mm (range: 15 to 90 mm). The mean operating time was 132 min (range: 70 to 240 mm) for an estimated mean blood loss of 120 cc (range: 0 to 1000 cc). The mean hospital stay was 8.3 days (range: 4 to 26 days). Conversion was necessary for technical difficulties in 7 cases (16.7%), venous injuries in 5 cases, arterial injury in 1 case, and difficult dissection of the perirenal fat due to a history of high-grade diffuse lymphoma in remission. The final histological stage showed 31 pT1 tumours, one pT2 tumour and 10 pT3 tumours. Four patients died in a context of metastatic diseases with a mean follow-up for our series of 15 months (range: 1 to 64 months). CONCLUSION: Laparoscopic retroperitoneal radical nephrectomy is a feasible technique, which strictly complies with the principles of cancer surgery. The main complication observed in our series was bleeding, which appears to depend on the surgeon's experience. PMID- 15822386 TI - [Results of open radical nephrectomy in a series of 230 patients]. AB - OBJECTIVE: To analyse the oncological results of open radical nephrectomy for renal cancer PATIENTS AND METHODS: Between December 1989 and June 2003, open radical nephrectomy for renal cancer was performed by the same operator in 230 patients. Cancers treated by partial nephrectomy were excluded from this study. The operation was performed via a lumbar incision in 95.7% of cases. Surveillance consisted of clinical examination and thoracic and abdominopelvic computed tomography twice a year for the first two years, then annually for 3 years, and then at 7 years and at 10 years in the absence of recurrence. RESULTS: With a median follow-up of 63.1 months, the 5-year and 10-year recurrence-free survival rates were 88.4% and 67.2%, respectively. Tumour size, tumour stage (TNM 1997), Fuhrman's nuclear grade, histological type of cancer, and the presence of suggestive signs were correlated with the recurrence-free survival rate. The 10 year recurrence-free survival rate was zero in patients with visceral or lymph node metastases at the time of diagnosis. The patient's age at the time of diagnosis and the presence of a paraneoplastic syndrome did not influence recurrence free survival in this series. CONCLUSIONS: This study confirms the data of the literature: tumour size, tumour stage, nuclear grade, histological type, and suggestive signs are prognostic factors of renal cancer. Radical nephrectomy ensures a global 5-year recurrence-free survival rate higher than 85%. PMID- 15822387 TI - [Complications of Bricker ileal conduit urinary diversion: analysis of a series of 246 patients]. AB - OBJECTIVE: The objective of this retrospective study was to evaluate the mortality and early and late morbidities of Bricker ileal conduit urinary diversion. PATIENTS AND METHODS: Between January 1990 and December 2002, 246 Bricker ileal conduit urinary diversion was performed in our centre in 164 males (67%) and 82 females (33%) with a mean age of 64 years (range: 9 to 90 years). Bricker diversion was performed in 73.6% of cases for underlying tumour (prostate, bladder, cervical, colon cancer), and in 26.4% of cases for benign disease (neurogenic bladder radiation bladder bladder exstrophy, incrusting cystitis). Cystectomy was associated with Bricker diversion in 62.2% of cases. The mean follow-up was 24 months (range: 1 to 151 months). The following parameters were studied: mortality and early and late medical or surgical, urological and gastrointestinal complications. RESULTS: The postoperative mortality was 6.9% (17 deaths, 16 in patients in whom Bricker diversion was performed for cancer). The early morbidity was marked by gastrointestinal complications (ileus, fistula, evisceration) in 46 cases (1.7%), 25 of which required re-operation. A medical complication was observed in 41 patients (16.7% of the series), responsible for 60% of the postoperative mortality (10 of the 17 deaths). A urinary fistula was observed in 7 patients (2.8%). The late morbidity consisted of parietal complications (incisional hernia, peristomal hernia, stricture of the stoma) in 18.3% of cases. Urological complications consisted of acute pyelonephritis (11%), ureteroileal stricture (4.9%) and urinary stones (4.9%). CONCLUSION: Bricker ileal conduit urinary diversion is associated with considerable mortality, especially in cancer patients. Early complications are essentially gastrointestinal, while late complications tend to be parietal or urological. PMID- 15822388 TI - [Oncological results of 117 consecutive radical prostatectomies]. AB - INTRODUCTION: The objective of this study was to evaluate the risk factors of biochemical recurrence in patients with prostate cancer treated by retropubic prostatectomy. MATERIAL AND METHODS: Retrospective analysis of 121 consecutive retropubic radical prostatectomies performed between January 1994 and December 1997. Biochemical recurrence was defined by PSA > or = 0.2 ng/ml. Biochemical recurrence-free survival rates were calculated by the actuarial method. Univariate and multivariate analysis of prognostic factors of biochemical recurrence was performed. RESULTS: The actuarial biochemical recurrence-free survival was 71% at 5 year and 63% at 7.5 years. Kaplan-Meier analysis did not reveal any difference for the risk of biochemical recurrence between patients with doubtful margins and those with positive margins. These two groups were therefore combined. Significant risk factors on univariate analysis were: pathological stage, grade of the predominant cell contingent on the specimen, margins, highest cell grade on biopsies, clinical stage, PSA level, perineural invasion and Gleason score. Significant risk factors for biochemical recurrence on multivariate analysis were: pathological stage, PSA > 10 ng/ml, the highest grade on biopsies, positive margins and perineural invasion. CONCLUSION: Doubtful margins and positive margins appear to be associated with the same risk of biochemical recurrence. The prognostic value of Gleason grade on the specimen is higher than that of the Gleason score. A high Gleason grade on biopsies is associated with a particularly poor prognosis. PMID- 15822389 TI - [A five-year experience in conformational radiotherapy in the treatment of prostate cancer. Evaluation of acute toxicity in 131 patients]. AB - PURPOSE: To evaluate the urinary and rectal toxicity secondary to 3D conformal radiotherapy for prostate cancer. MATERIAL AND METHODS: Between 1998 and 2003, 131 men with prostate cancer underwent 3D conformal radiotherapy with or without androgen deprivation. The different stages were: 2 T1b ; 40 T1c; 19 T2a; 16 T2b; 18 T2c; 33 T3a; 1 T3b and 2 T3c with Gleason score: 4-6 = 47%, 7 = 36% and 8-9 in 17% of the cases. The median patient age was 66 (48-87). Pretreatment PSA level was respectively < 10 ng/ml (41%). 10-20 ng/ml (30%) and > 20 ng/ml (29%). Of the 131 patients, 98 received androgen ablation therapy before radiation. The total radiation dose varied between 66 and 74 Gy, delivered with 18MV photons of the linear accelerator, the median follow up was 33 months (5-67). RESULTS: According to the RTOG grading (gr) for acute toxicity, we noticed 3gr 3 genitourinary (GU) toxicity and no gr3 gastro intestinal (GI) toxicity. There were 36 gr 1 and 12 gr 2 GI toxicity, 41 gr 1 and 22 gr 2 GU toxicity. The mean prostate volume was 41 cc for patients who received androgen ablation and 56 cc for the others (p < 0.002). The percentage of volume receiving more than 50 Gy (V50) was calculated, the median V50 was 32% (5-67) for the rectum and 35% (5-79) for the bladder CONCLUSION: The toxicity profile in this study is in the same range than those of the literature and of our previous study concerning our first 50 patients with prostate cancer treated with 3D conformal radiotherapy. PMID- 15822390 TI - [Management of acute prostatitis, based on a series of 100 cases]. AB - OBJECTIVES: To evaluate the current diagnostic and therapeutic management of acute prostatitis. MATERIAL AND METHODS: The authors report a series of 100 consecutive patients with a diagnosis of acute prostatitis managed between January 1999 and December 2003. They analysed the clinical and laboratory data and imaging findings leading to the diagnosis of acute prostatitis and then the modalities of treatment and follow-up of these patients. RESULTS: The median age was 56.5 years (range: 19-86 years). In 77% of cases, the initial septic syndrome was considered to be severe. Prostatic tenderness was detected on digital rectal examination in 68% of cases. In 76% of cases, the micro-organism responsible was Escherichia coli. 8% of patients had positive blood cultures. All patients presented laboratory signs of inflammatory syndrome, with elevated C Reactive Protein (CRP) in most cases. Renal and bladder ultrasound was performed in 49% of cases and CT was performed in 16% of cases. 92% of patients were treated with fluoroquinolones, as monotherapy in 34% of cases. The duration of prescription ranged from 2 weeks to 6 weeks. No aetiology was detected in 48% of cases, 8% were considered to be iatrogenic and 44% were associated with incomplete bladder emptying. CONCLUSION: Acute prostatitis, rapidly treated by aggressive and adapted antibiotics has a rapidly favourable outcome. Elevation of CRP and the presence of leukocyturia on urine dipsticks are almost constant and, when they are normal, the diagnosis of acute prostatitis should be questioned. Imaging is not required in the absence of diagnostic doubt and any suspicion of progression to abscess. The duration of antibiotic therapy varies from prescriber to prescriber reflecting the various guidelines. Guidelines on this subject are contradictory and further effort is required to achieve homogenization and application of these guidelines. PMID- 15822391 TI - [Quality of life assessment after TVT (tension-free vaginal tape) based on 3 different questionnaires]. AB - OBJECTIVE: To prospectively evaluate the effects of TVT on the quality of life of patients with urinary incontinence by using 3 different questionnaires. PATIENTS AND METHODS: Fifty one patients with a mean age of 57.1 years (range: 34 to 82) were treated by TVT for urinary incontinence combined with laparoscopic sacral colpopexy for associated cystocele in 12 cases. Fifty of these patients suffered from stress urinary incontinence and one presented mixed urinary incontinence. These patients completed 2 quality of life questionnaires and a urinary symptom score: Ditrovie and Contilife scale and the MHU scale for measurement of urinary handicap before and 3 months after TVT. Statistical analysis was based on Student's test. RESULTS: The mean scores obtained with the MHU scale were 6.74 (range: 2-16) preoperatively and 1.4 (range: 0-7) postoperatively. The mean score obtained with the Ditrovie scale was 3.1 (range: 2-4) preoperatively and 1.32 (range: 1-3) postoperatively. The mean score obtained with the Contilife scale was 82.1 (range: 51-119) preoperatively and 38.8 (range: 30-15) postoperatively. The difference was statistically significant (p<0.05) for the MHU and Contilife scales, but not for the Ditrovie scale. CONCLUSION: TVT allows a marked improvement of the quality of life of patients with urinary incontinence. The MHU and Contilife questionnaires can be used to objectively quantify the improvement of quality of life after TVT and, due to their marked and reproducible variation, they can also be used to assess the degree of improvement and the stability of the results over time. PMID- 15822392 TI - [Pudendal nerve block: local anaesthesia for insertion of suburethral tape: preliminary study of 9 cases]. AB - PURPOSE: The objective of this study is to evaluate the safety of local anaesthesia by pudendal nerve block (PNB) for insertion of TVT or TOT suburethral tape for the treatment of female urinary incontinence. MATERIAL AND METHOD: PNB is performed by bilateral injection between the anus and ischium. The pudendal nerve, detected by electrostimulation, is anaesthetised by a solution of Naropeine and Carbocaine. Anaesthesia is systematically completed by abdominogenital and genitofemoral nerve blocks and by deep infiltration in the vagina. Perioperative and postoperative pain was assessed by visual analogue scale (VAS). RESULTS: Nine patients were treated by suburethral tape under PNB between November 2002 and January 2003. The mean age was 65.5 years (range: 46 to 87 years). TVT was used in 6 cases and TOT was used in 3 cases. PNB was well tolerated by the patient in 66% of cases. Eight patients (88%) did not experience any pain during the operation. The majority of patients (88%) were very satisfied with the anaesthesia. One patient experienced residual pain for 3 weeks. The technique is easy to perform by the anaesthetist and did not modify the surgical technique. This technique is relatively inexpensive. CONCLUSION: This preliminary study on a limited number of cases shows that PNB appears to be a good alternative to the usual anaesthesia techniques for insertion of suburethral tape. PMID- 15822393 TI - [Gestational diabetes and urinary tract malformations: a hospital case-control study]. AB - INTRODUCTION: Urinary tract malformations represent the leading cause of renal failure in children. Gestational diabetes plays a controversial role in the development of urinary tract malformations. The purpose of this study was to assess the role of insulin-requiring gestational diabetes on the development of ureteric malformations in children. MATERIAL AND METHODS: The authors report the results of a prospective case-control survey (102 cases, 104 controls) conducted in a teaching hospital. The role of insulin-requiring gestational diabetes on the development of urinary tract malformations was tested by logistic regression after adjustment for risk factors for urinary tract malformations 'family history of urinary tract malformations, parity, smoking, substance abuse during pregnancy). RESULTS: Gestational diabetes is a risk factor for urinary tract malfornations after adjustment (aOR=5.1; 95%CI: 1.1-24.5), and for ureteropelvic junction syndrome (aOR=6.5; 95%CI: 1.1-39.4) and vesicoureteric reflux (aOR=5.9; 95%CI: 1.1-32.1). CONCLUSION: All perinatal healthcare professionals must be familiar with the concept of insulin-requiring gestational diabetes as an uncommon, but fairly powerful risk factor for ureteric malformations. PMID- 15822394 TI - [Interest of mobilization of the urethral plate in the release of chordee related to posterior hypospadias]. AB - OBJECTIVE: To compare the advantages of release of the skin and dartos to those obtained after mobilization of the urethral plate and resection of the underlying fibrous tissue (according to Mollard). MATERIAL AND METHODS: Between February 1996 and March 2003, 97 patients were operated for posterior hypospadias. Lateral photographs were taken in these patients during successive erection tests: a first test was performed at the beginning of the operation, a second test was performed after release of the skin and dartos, a third test was possibly performed after mobilization of the urethral plate and resection of the underlying fibrous tissue, and a last test was performed at the end of the operation. RESULTS: Release of chordee with preservation of the urethral plate was achieved in 88 patients (91% of cases). Analysis of the lateral photographs showed that dissection of the skin and dartos ensured correction of chordee less than 55 degrees, while this procedure was insufficient for chordee exceeding 90 degrees. Dissection of the urethral plate and resection of the underlying fibrous tissue only allowed correction of chordee between 10 degrees and 25 degrees, with no increase of the postoperative complication rate. CONCLUSIONS: Correction of chordee associated with posterior hypospadias is essentially obtained by release of the skin and dartos and mobilization of the urethral plate does not endanger the blood supply. PMID- 15822395 TI - [Neonatal uro-haematoma secondary to posterior urethral valve]. AB - The authors report the case of a male neonate presenting with a heterogeneous cystic mass in the right renal area with no identifiable renal unit. The diagnosis of right perirenal extracapsular uro-haematoma secondary to posterior urethral valve was proposed on the basis of the antenatal clinical history. The late discovery at 39 weeks of amenorrhoea (pregnancy not followed) of a foetus with trabeculated bladder associated with left ureteropelvic dilatation, and a cystic mass occupying the right lumbar fossa was highly suggestive of right urinoma secondary an infravesical obstacle. In this case, vaginal delivery was probably responsible for intracystic bleeding of the urinoma. The postnatal morphological assessment including ultrasound, retrograde cystourethrography and MRI confirmed the presence of a posterior urethral valve complicated by right uro haematoma displacing the right kidney and left ureteropelvic dilatation. This lesions corresponds to a collection of urine and blood in the right perirenal space due to caliceal or parenchymal perforation secondary to raised urinary tract pressure above an obstacle. Subcapsular or extracapsular perirenal urinoma, sometimes detected in neonates with posterior urethral valve, reflects protective decompression of the urinary tract (safety valve effect), as in our case. The neonatal diagnosis of a heterogeneous renal cystic mass is sometimes difficult (polycystic renal dysplasia, cystic lymphangioma, cystic neuroblastoma, adrenal haematoma). In some cases, the antenatal history may suggest the diagnosis of perirenal urohaematoma secondary to infravesical obstruction. PMID- 15822396 TI - [Bladder melanoma in a 7-year-old girl]. AB - Melanomas are very rare in children. The authors report an original case of bladder melanoma in a 7-year-old girl. The initial staging assessment and complete remission 7 years after partial cystectomy with healthy resection margins confirmed the primary nature of this tumour Bladder melanomas are exceptional in adults, as only about 20 cases have been published. To the best of our knowledge, this is the first published case in a child. PMID- 15822397 TI - [Laparoscopic kidney release for refractory chyluria]. AB - Chyluria is a rare disease outside of parasite endemic zones. The treatment of severe forms is surgical. Reno-lymphatic disconnection is the reference procedure, with long-term success rates of 99%. This operation performed via a lumbar incision can also be performed by retroperitoneal or transperitoneal laparoscopy. The advantages of this minimally invasive incision are those classically reported in the literature. PMID- 15822398 TI - [Idiopathic acute partial renal infarction]. AB - Renal infarction is a rare disease which must be considered in any case of low back pain. The most frequent causes are related to emboligenic heart disease, renal artery fibrodysplasia or trauma. The authors report the second published case of idiopathic renal infarction. The diagnosis is usually suggested by contrast-enhanced abdominal CT scan, but is confirmed and refined by selective renal arteriography or CT angiography. Treatment may be medical or surgical depending on the severity of the lesions and the patient's general state. The therapeutic indications are discussed in the light of a review of the literature. PMID- 15822399 TI - [Benign schwannoma, a rare retroperitoneal tumour]. AB - Primary retroperitoneal tumours are rare in adults. Among retroperitoneal tumours derived from nervous tissue, schwannoma is defined as a tumour arising from neurilemma or Schwann nerve sheath cells. It is a very rare tumour, especially in the retroperitoneal site (0.2%). Its diagnosis is immunohistological, and treatment is surgical, consisting of complete resection of the mass due to the risk of malignant degeneration. The authors report the case of a 38-year-old patient admitted with isolated right lumbar pain. Imaging (ultrasound and CT urography) revealed a retroperitoneal tumour measuring 6 x 9 cm. Resection was performed by right thoraco-pleuro-laparatomy. The immunological and histological study concluded on benign schwannoma. With a postoperative follow-up of two years, the patient has not presented any recurrence of this retroperitoneal schwannoma. PMID- 15822400 TI - [Pelvic lipomatosis associated with glandular cystitis. Report of two cases]. AB - The authors report two cases of pelvic lipomatosis associated with glandular cystitis. This is a rare disease with variable and nonspecific clinical features. Complementary investigations useful for the diagnosis of pelvic lipomatosis are CT and especially MRI, intravenous urography and biopsies. The clinical features, radiological findings and therapeutic approach are discussed in the light of a review of the literature. PMID- 15822401 TI - [Orbital metastasis of prostatic carcinoma]. AB - Prostate cancer is frequently complicated by bone metastases. However some metastatic sites remain rare and unusual. The authors report an exceptional case of prostatic carcinoma presenting with orbital metastasis. A 71-year-old man was admitted with ocular protrusion and decreased visual acuity of the left eye. In addition to the ophthalmological disorders, clinical examination revealed a left temporoparietal swelling with a suspicious prostate on digital rectal examination. Brain MRI revealed an osteolytic process involving the left sphenoid bone with intra-orbital invasion. Another osteolytic site was observed in the left temporoparietal region with intracranial and extracranial components. PSA was 13,327 ng/ml. Prostatic biopsy confirmed the diagnosis of prostatic carcinoma. Treatment comprised cranial radiotherapy combined with anti-androgen endocrine therapy and corticosteroids. Six weeks after treatment, the patient obtained an improvement of visual acuity and regression of exophthalmos and the temporoparietal swelling. Orbital metastasis of prostatic carcinoma raises the problem of ophthalmic emergency. Cranial radiotherapy and anti-androgen endocrine therapy allowed regression of the symptoms without modifying the unfavourable course of the disease. PMID- 15822402 TI - [Urethral cancer in African women]. AB - Urethral carcinoma is a rare tumour. The authors report the first three cases diagnosed in black African woman. The diagnosis is difficult but can be much more seen if frequently through of. Urethral smear for cytology and histology are essentials elements in the diagnosis. Surgical treatment of this tumour is cheap and quite accessible, hence the need for early diagnosis. The prognosis is poor even with combined surgical and radiotherapy. PMID- 15822403 TI - [Postoperative ureterocolonic fistula: a case report]. AB - Postoperative ureterocolonic fistula is exceptional: only one case has been published over the last 32 years. The authors report a case of postoperative ureterocolonic fistula observed after right hysteroadnexectomy in a 38-year-old woman with a solitary right kidney. This patient initially presented with anuria and fever followed by urine-smelling liquid diarrhoea. In a context of inadequate urological imaging equipment, the diagnosis of ureterocolonic fistula was suspected on the basis of the specific features of the diarrhoea, ultrasound which revealed ureterohydronephrosis above a pelvic obstacle, and normal cystography. Surgical exploration demonstrated a ureterosigmoidfistula. This case represents the 20th case of ureterocolonic fistula reported in the literature, including 2 postoperative cases. PMID- 15822404 TI - [KIT receptor in testicular seminoma]. AB - KIT receptor expression is observed in the majority of seminomas. Activation of KIT tyrosine kinase due to somatic mutations has been demonstrated. Mutations of the c-kit gene in testicular seminomas are located in exon 17. Inhibitors of KIT tyrosine kinase activity can have a therapeutic role, particularly in seminomas with a c-kit mutation sensitive to imatinib mesylate. A clinical trial plans to examine the efficacy of imatinib mesylate in the treatment of metastatic seminomas refractory to chemotherapy. Tyrosine kinase inhibitors can also be tested in patients with minimal retroperitoneal lymph node involvement before radiotherapy. If they are active, future therapeutic trials could include the use of these inhibitors as adjuvant therapy for patients with stage I seminoma in order to decrease the potential risk of second tumour. PMID- 15822405 TI - [How to identify the neck of a caliceal diverticulum in flexible ureterorenoscopy]. AB - A caliceal diverticulum corresponds to a cavity communicating with the renal excretory tract via a neck. Surgical treatment is required when the caliceal diverticulum is symptomatic with repercussions on the patient's quality of life. A retrograde approach via holmium:YAG laser ureterorenoscopy has been recently used to treat the stone and externalize the caliceal diverticulum into the pyelocaliceal cavities. The problem raised by this approach is identification of the diverticular neck. The objective of this technical note is to describe a technique to facilitate identification of the diverticular neck. PMID- 15822406 TI - [Prospective study of iliac extraction of the kidney after laparoscopic nephrectomy]. AB - INTRODUCTION: Various routes are used to extract the operative specimen during laparoscopic nephrectomy. The main points are compliance with rules of cancer surgery, when applicable, and minimum wall destruction. The objective of this study was to prospectively evaluate the low iliac approach. MATERIAL AND METHOD: Prospective follow-up of 23 laparoscopic nephrectomies (17 tumours and 6 living donor kidney harvestings) in which the kidney was extracted via a low iliac incision measuring 5 to 7 cm. RESULTS: The mean operating time was 188 +/- 50 min with a mean blood loss of 112 +/- 126 ml. The mean duration of the incision was 10 minutes. In the case of a tumour the mean weight of the operative specimen was 571 +/- 127 g in and the mean diameter of the mass was 5.7 +/- 1.9 cm. All grafted kidneys functioned normally. No surgical conversion was necessary. The mean follow-up was 9.6 +/- 1.2 months. No late postoperative complications were observed. CONCLUSION: Radical nephrectomy or living donor kidney harvesting can be performed via laparoscopy with extraction of the operative specimen via a low iliac incision. This incision ensures extraction of very large specimens while preserving the aesthetic and functional advantages of laparoscopy with no increased cancer risk. They are simple to perform and easily reproducible via a transperitoneal or retroperitoneal approach. Absence of muscle section maintains the integrity of the abdominal wall. No postoperative incisional hernia has been observed. PMID- 15822407 TI - [Contribution of Etienne-Frederic Bouisson (1813-1884) to the surgical treatment of hypospadias]. AB - Professor Bouisson was a great surgeon and benefactor of medicine. He exercised successfully the functions of dean, rector and deputy. A great part of his surgical work deal with pediatric urology. Bouisson's dissertation on surgical treatment of hypospadias constitute a crucial turning-point in the history of treatment of this malformation. PMID- 15822408 TI - [Physiology of the anal and urinary sphincter apparatus for continence]. AB - This report of the 27th congress of the Societe Internationale Francophone d'Urodynamique (SIFUD) describes the mechanisms which allow the anal and urethral sphincter apparatus to ensure faecal and urinary continence. It successively described sphincter anatomy, the structure and ultrastructure of its muscle components, its innervation, neurotransmission and functioning regimens at rest, on effort and on straining. This study illustrates the differences but also the similarities of functioning of these two sphincter apparatuses. This can explain the high frequency of associated urinary and anal incontinence. PMID- 15822409 TI - The legality of liability surcharges. PMID- 15822410 TI - What's in your contract? Why it's more important than ever to know. PMID- 15822411 TI - Answers to questions about fees for copying medical records. PMID- 15822412 TI - Physicians protected from termination during blues lawsuit. PMID- 15822413 TI - Finding meaning--what is expected of physicians. PMID- 15822414 TI - Trauma during pregnancy: a situation pregnant with danger. AB - Trauma in pregnancy is currently a leading cause of non-pregnancy-related maternal death, and maternal death remains the most common cause of fetal demise. The most common etiologies of trauma in pregnancy include transportation accidents, falls, violent assaults, and burn injuries. Head and neck injuries and hemorrhagic shock account for most maternal deaths secondary to trauma. Women of childbearing age are among the population at greatest risk for trauma. The pregnant trauma victim presents a unique spectrum of challenges to the health care team. Expeditious maternal resuscitation is the most effective method of fetal resuscitation. The management of pregnant trauma victims requires the anesthesiologist, the obstetrician and the trauma surgeon to consider and understand the unique changes in anatomy and physiology that take place during pregnancy. This article reviews the current considerations for the optimal perioperative management of pregnant trauma victims. PMID- 15822415 TI - Herbal ecstasy: cardiovascular complications of khat chewing in pregnancy. AB - Chewing fresh leaves of the Khat plant (Catha edulis Celestrasae) is a widespread habit (also practiced by women, even during pregnancy) with a deep-rooted tradition in East Africa and the Arabian Peninsula. With the influx of immigrants from East Africa and the Arabian Peninsula khat chewing has been imported into other countries including Europe the United States. The major pharmacologically active constituent of the fresh khat leaves is cathinone. Khat (also known as herbal ecstasy) is chewed for its central nervous system stimulant properties, which resemble amphetamine. Cardiovascular complications from cathinone use may therefore be similar to those of amphetamine. I herein present the first reported case of a pregnant patient who developed chest pain, tachycardia, and hypertension following khat-chewing session. PMID- 15822416 TI - Once a post-dural puncture headache patient--always post-dural puncture headache patient: an update. PMID- 15822417 TI - Continuous interscalene block using a stimulating catheter: a review of the technique. AB - The management of postoperative pain after major shoulder surgery can be achieved successfully with a continuous interscalene block. This article reviews the essentials of the stimulating catheter technique for the continuous interscalene block that was described by Boezaart in 1999. The authors also describe their experience and results with the first two hundred catheters they placed. PMID- 15822418 TI - Preliminary results of prolonged target controlled infusion of sufentanil adjusted to an effort pain score after cardiac surgery. AB - Repeated boluses of i.v. morphine are often used for analgesia after cardiac surgery, but this procedure frequently provides inadequate pain relief. Target controlled infusion (TCI) of opioid drugs has been proposed as an alternative. The objective of this study was to evaluate the effects of prolonged sufentanil TCI adjusted to an effort pain score on the postoperative course after cardiac surgery. Twenty-six patients scheduled for cardiac surgery were anaesthetised using TCI of propofol and sufentanil, followed by early extubation. In the postoperative period, patients were randomly allocated to receive either boluses of i.v. morphine (Bolus group), adapted to a pain score at rest, or sufentanil TCI with a low target concentration (0.08-0.1 ng/ml), adapted to a pain score during deep inspiration (TCI group). Postoperative pain was assessed using a Verbal Numerical Score (VNS) before and during three stimuli: extubation, lateral positioning and physiotherapy. In 15 patients, inspiratory capacity was evaluated by incentive spirometry (IS) on postoperative day 1. Three patients in each group were withdrawn because of delayed extubation. Clinical characteristics, mean time to extubation and PaCO2 were similar in both groups. Analgesia was adequate at rest in both groups as indicated by low pain scores. Pain intensity increased significantly during the three stimuli (p < 0.01). VNS were lower in the TCI than in the Bolus group (3.8 +/- 0.5 versus 4.8 +/- 0.5, p = 0.03). Nine of 10 patients in the TCI group but only 4 of 10 patients in the bolus group gave a VNS below 5 during the stimuli. IS performance was better preserved in the TCI than in the Bolus group (53 +/- 5% versus 35 +/- 5% of preoperative values, p < 0.05). These observations indicate that after cardiac surgery, postoperative pain management with prolonged TCI of sufentanil adapted to a pain score during deep inspiration can achieve better analgesia during routine bedside procedures and higher pulmonary volumes than on-demand boluses of morphine. PMID- 15822420 TI - Remifentanil for obstetric analgesia and anesthesia: a review of the literature. PMID- 15822419 TI - Effects of nitrous oxide on spectral entropy of the EEG during surgery under balanced anaesthesia with sufentanil and sevoflurane. AB - BACKGROUND: Spectral entropy of the electroencephalogram (EEG) has been proposed to monitor anaesthetic depth. We investigated the effect of nitrous oxide on response (RE) and state entropy (SE) of the EEG during lumbar disc surgery under anaesthesia with sufentanil and sevoflurane. METHODS: In an open study, anaesthesia was induced with propofol and sufentanil, and maintained with 2% end tidal sevoflurane concentration in air/oxygen (FiO2 = 0.4) in 25 patients. During surgery, nitrous oxide was randomly administered either at 0 or at 60% end-tidal concentration in 10 (control group) and 15 patients (nitrous oxide group), respectively. RE and SE were recorded at 2.5 min intervals for 10 min before randomization and for 25 min either continuously (control) or after achieving the target nitrous oxide concentration. RESULTS: Two patients who received nitrous oxide were excluded from statistical analysis because of protocol violation. Nitrous oxide provoked a significant decrease in RE and SE from 46.2 +/- 11.1 and 44.3 +/- 11.1 to a lowest value of 27.8 +/- 8.3 and 27.1 +/- 8.9, respectively. The decrease in entropy persisted during the 25 min recording period. CONCLUSIONS: Addition of nitrous oxide during balanced anaesthesia with sufentanil and sevoflurane provokes a decrease in response and state entropy of the EEG during lumbar disc surgery. PMID- 15822421 TI - The lure of hypocrisy. AB - Why are we susceptible to hypocrisy? Four preconditions for falling prey to hypocrisy are delineated. The reasons for our complex attitude to hypocrisy, both condoning and condemning it, are also explored. Hypocrisy is the false claim to virtue. It always refers to consciously intended deception by a person in a position of trust. Making use of literary examples, the investigation starts with the delineation of three readily apparent preconditions for falling prey to hypocrisy. Idealization of the hypocrite is seen as a defense against a dread of uncertainty on the part of the person who succumbs to hypocrisy. The addition of a third precondition, the force of powerful desire, completes the introduction. A selective review of historical and philosophical studies of hypocrisy over the past twenty-five hundred years situates the problem of the susceptibility to hypocrisy. Application of psychoanalytic formulations and clinical data, principally from the study of patients whose psychoanalysts committed ethical violations, produces a fourth precondition for succumbing to hypocrisy: transference, with a regression from mature trust to a condition of unmodified basic trust. PMID- 15822422 TI - Intuition: a bridge to the coenesthetic world of experience. AB - The concept of intuition is relatively unestablished in psychoanalysis, where it is often associated with narcissistic meanings and vagueness. But intuition, as an integrated mode of archaic coenesthetic thinking, should be kept conceptually free of those connotations. Its capacity of undifferentiated delineation supplies an instinctive general means of dealing immediately with various rationally indistinct phenomena, such as forms, shades, and multidimensionality, regardless of the boundaries between sensory modalities. It may be impossible to translate intuitive experiences into lexical form; these languages are incommensurable. Intuition as a preconscious nondiscursive thinking process is needed in creativity, as well as less conspicuously in countless everyday activities. In speech communication, intuition rapidly specifies subtle shades of meaning in linguistic content and all the prosody. In psychoanalytic work intuition is like radar, creating preliminary contacts with the inner world of the analysand. The observations gained require, however, rational consideration to be confirmed. Intuition is an essential instrument of the psychoanalyst, and also functions in the service of tact to create working space and adequate forms of interpretations. Clinical vignettes reflecting some problematic fates of special intuitiveness in creativity are presented from psychoanalytic work with artists. PMID- 15822423 TI - The evolution of paternal identity in late adulthood. AB - The experience of fatherhood in late adulthood has received little attention in the psychoanalytic literature. After a presentation of the literature on fatherhood and a consideration of the developmental tasks of late adulthood in which the experience of fatherhood is embedded, the dynamically charged, developmentally conflicted nature of the relationships between late-life fathers and their children is explored. A basic premise is that the changing nature of these relationships inevitably confronts the elderly patriarch with considerable intra-psychic turmoil, reflection, and conflict that must be engaged and mastered if these most significant relationships are to be maintained and sustained for both generations. Clinical examples illustrate the theoretical points presented. PMID- 15822424 TI - Rethinking desire: the objet petit a in Lacanian theory. AB - The concept of the objet petit a is central to Lacan's theory of desire, which arguably represents his major contribution to psychoanalysis. It is an expression of the lack inherent in human beings, whose incompleteness and early helplessness produce a quest for fulfillment beyond the satisfaction of biological needs. The objet petit a is a fantasy that functions as the cause of desire; as such, it determines whether desire will be expressed within the limits of the pleasure principle or "beyond," in pursuit of an unlimited jouissance, an impossible and even deadly enjoyment. Parallels between the objet petit a and Winnicott's transitional object are explored and its functions illustrated through analysis of Pedro Almodovar's film Talk to Her. A clinical case is presented in which the question of desire seemed crucial. PMID- 15822425 TI - Writing about patients: IV. patients' reactions to reading about themselves. AB - When analysands read about themselves in reports, their reactions range from anger, disappointment, or condemnation to a sense of appreciation or even idealization of the analyst. The eleven interviews reported here reflect only conscious responses; the unconscious layers were not probed for. It should be kept in mind also that the analysts of these patients might report very different stories. Other limitations are the small sample size and the representation only of patients who volunteered. Nonetheless, the information they provide may help analysts consider how and when writing about patients may influence their representation of themselves, the analyst, and analysis itself. PMID- 15822426 TI - Writing about patients: V. analysts reading about themselves as patients. AB - The narratives of twenty analysts written about when they were patients are presented. Their stories provide suggestions about practices to avoid when writing clinical material, but no generalized prescriptions emerge. Individualization and sensitivity to the situation for each pair remain the best guide. The experiences these analysts recount run the gamut of emotions, from negative through neutral to positive. The neutral responses came mainly from twelve analysts who in the course of an interview about their own writing told of having been written about as patients. The other eight, volunteers who initiated contact for the sole purpose of reporting their experience of having been written about, appear on average to be motivated by stronger affective reactions. The era in which the analyst wrote also seems to have influenced the reactions; in earlier times, not asking permission was accepted professional practice. Today, however, it is increasingly common to ask permission when extended clinical examples are published. One problem specific to analyst-patients was concern about the loss of their role as patient when their analyst engaged them collaboratively in the writing. PMID- 15822427 TI - Affects, agency, and self-regulation: complexity theory in the treatment of children with anxiety and disruptive behavior disorders. AB - In an increasingly unsettled and violent world, with swelling numbers of children who are abused, abandoned, or neglected, emotionally if not physically, and an increasing population of aggressive preschool children with anxiety and disruptive behavior disorders who cannot be contained in ordinary settings, psychoanalysts can make a contribution. Early intervention is essential. In very early childhood, new procedural memories for interacting with others and for regulating affects can be formed more easily than they can ever be again. Intervention should aim toward helping the child develop a sense of agency, establish moral standards, assume self-responsibility, and attain the capacity for emotional regulation. The principles of complex dynamic systems can inform psychoanalytic treatment strategies, as demonstrated with five children whose cases are presented. PMID- 15822428 TI - Translating child analysis from the playroom to the classroom: opportunities and choices. AB - Over the years, psychoanalysts and educators have made efforts to meld a psychoanalytic orientation with the education of young children. The resulting programs have variously been called psychoanalytic nurseries, therapeutic nurseries, therapeutic preschools, or psychoanalytic early childhood programs. The methodology of the application or integration of psychoanalysis with the education of young children has received relatively little attention. After a brief historical review of the application of psychoanalysis to helping children in group educational settings, a number of features common to programs integrating psychoanalysis and early childhood education are described. A specimen program is next presented in which an application of psychoanalysis is used to assist children whose development is proceeding in a psychopathological direction. Also demonstrated are various ways in which psychoanalysis may be adapted to such programs, some options involved, and the capacity, realized and potential, for work within a therapeutic nursery to expand the field of inquiry and the therapeutic action of child analysis as conducted within a traditional framework. PMID- 15822429 TI - Child psychoanalysis and obsessive-compulsive symptoms: the treatment of a ten year-old boy. AB - Obsessive-compulsive symptoms have long held interest for psychoanalysts. Currently, pharmacological and behavioral treatments are commonly viewed as the standard of care, and psychoanalytic clinicians are often uncertain of how best to approach these patients. The successful two-year psychoanalytic treatment of a ten-year-old boy with obsessivecompulsive disorder, who remained symptom-free eight years after termination, is reported. While this is but a single case, its outcome implies that for some patients psychoanalysis may be the treatment of choice. PMID- 15822430 TI - Early developmental issues reconsidered: commentary on Pine's ideas on symbiosis. AB - The concept of symbiosis is a powerful one expressing a significant emotional experience for many individuals. However, the concept needs to be differentiated from infant-stage symbiosis, Margaret Mahler's second stage of normal development. The latter idea, according to Pine (2004), was of "central" importance to Mahler. Although Pine acknowledges the influence of research findings and some critical challenges to this idea, he maintains nonetheless that it "remains quite valuable" and reports what he suggests are manifestations of symbiotic experiences in the infant-mother relationship. His recognition of infant observational data has led him to feature the idea of "moments" of merger between mother and infant. Empirical data, however, challenge both the idea of a more broadly defined stage of infantile symbiosis and the more narrowly conceived "moments." A different understanding of development in the presence of early stage symbiosis is provided. PMID- 15822431 TI - Storytelling and psychoanalytic diversity. PMID- 15822432 TI - Postanalytic contacts in institutes. PMID- 15822433 TI - The lawsuit and democratic procedures. PMID- 15822434 TI - Constructive dialogue and editorial policy. PMID- 15822435 TI - Rapaport, Rangell, and general theory. PMID- 15822436 TI - [CME radiology 6. Acute dyspnea and productive cough]. PMID- 15822437 TI - [What is your diagnosis? Arteria lusoria]. PMID- 15822438 TI - [Contraception and bleeding disorders--diagnostic contributions by ultrasound]. AB - Practically all widely used and safe and reversible methods of contraception exert a direct (or at least an indirect) impact on the endometrium. Not surprisingly, abnormal uterine bleeding is a frequent adverse reaction associated with contraception. In this situation sonography--in particular transvaginal sonography--offers the physician a practically non-invasive imaging technique for determining the etiology of abnormal uterine, cervical or vaginal bleeding. Ultrasonographic imaging provides the physician with a detailed picture of the internal reproductive organs. It can also be used to monitor the efficacy of the particular contraceptive method. e.g. to check the position of IUDs, confirm the suppression of ovarian follicle activity in women taking ovulation inhibitors, detect ovarian cysts as a focus of undesirable hormonal activity, etc. In some cases, the failure of contraception, namely pregnancy, can be diagnosed by ultrasound. By using high-frequency linear ultrasonographic probes, the physician can verify the position of subcutaneous contraceptive implants on the medial upper arm. Therapeutic consequences can be derived, in particular, from the ultrasonographic measurement of the total thickness of the endometrial layer. PMID- 15822439 TI - [Evaluation of cervical lymph nodes by ultrasound]. AB - Abscesses and calcifications in cervical lymph nodes can be easily detected by ultrasound. Lymphomas often manifest in cervical lymph nodes and change size and sonografic structure. Lymph node metastasis of head and neck carcinomas are characterised by size, shape, vascularisation and infiltration. Sonografic criterias are discussed, especially to identify metastatic disease. PMID- 15822440 TI - [Sonographical differential diagnosis of cervical lumps]. AB - A brief synopsis for a practical approach in the clinical and sonographical evaluation of cervical lumps except for lymph node diseases is presented. PMID- 15822441 TI - [54-year-old patient with type 2 diabetes mellitus and increased blood pressure values]. PMID- 15822442 TI - [CME--EKG. 4. Solution. Lost competitive race. AV nodal reentrant tachycardia]. PMID- 15822443 TI - [Rationalist, magician, scharlatan? Samuel Hahnemann and homeopathy from the viewpoint of homeopathy]. PMID- 15822444 TI - [Non-invasive blood pressure measurement in dogs and cats]. AB - Bloodpressure measurements are of interest for todays veterinary practice. It can be used for screening of incoming patients, the emergency case, the intensive care patient and sedated animal. Furthermore, like in human medicine, blood pressure can be influenced by several diseases. The most reliable method of arterial blood pressure measurement is the direct or invasive method. Only the non-invasive methods doppler and oscillometric methods are used in veterinary medicine. The article describes the method of measurement, and provides a review of the literature. The oscillometric method especially in dogs. Both methods measure a lower bloodpressure if compared with an invasive method. The difference in bloodpressure is for the oscillometric method in cats greater then in dogs and appears to increase at higher blood pressures. Keeping these facts in focus, and if one is able to work in a quiet environment, repeat the measurements and maintain a critical attitude, blood pressure measurements can be performed in veterinary practice. PMID- 15822445 TI - [Toes of the dog and owner]. PMID- 15822446 TI - [Drug resistance makes new control measures of stomach parasites in small ruminants necessary]. AB - Control of parasitic gastroenteritis in small ruminants is threatened by the worldwide growing problem of anthelmintic resistance. Therefore, alternative approaches for worm control are imperative. Of utmost importance is to slow down selection pressure for anthelmintic resistance by using alternative control measures combined with minimal anthelmintic usage. New control options for the Netherlands are discussed. PMID- 15822447 TI - [Life in a clean environment: risky if we have to believe the 'hygiene hypothesis']. PMID- 15822448 TI - [Dr. Marc Sprenger, Director-General RIVM. Interview by Paul Overgaauw]. PMID- 15822449 TI - [Rectal tears]. PMID- 15822450 TI - [Instruments, books and other objects memorable to (almost) forgotten opinions, therapies, buildings, et cetera. Original 'True Type' animal models of a Holstein Friesian cow and steer]. PMID- 15822451 TI - [Continuing education! Continuing education?]. PMID- 15822452 TI - [Puppy feed]. PMID- 15822453 TI - [Psychiatric and mental health plan. Strong gesture of acknowledgement or simple announcement effect?]. PMID- 15822455 TI - [Recommendations for the discharge of the hospitalized patient with evolving dependence]. PMID- 15822458 TI - [Understanding and caring in limited states. Interview by Dominique Barbier]. PMID- 15822459 TI - [Home care services, an opportunity for community psychiatric care]. PMID- 15822460 TI - [Home care services, the nurse at the heart of patient management]. PMID- 15822461 TI - [Home visits as therapeutic injunction and the law as mediation of care]. PMID- 15822462 TI - [Observation of babies at home]. PMID- 15822463 TI - [Home care services in the framework of therapeutic family admission]. PMID- 15822464 TI - [Drug monitoring and nursing practice]. PMID- 15822465 TI - [Ethical implications of clinical trials in Tunisia]. AB - Clinical trials are necessary for medical advancement. They must respect legal obligations. Ethical questions related to protection of the human being's rights are yielded by these trials. Joining research to medical core is problematical in consideration of patient's consent to clinical trial. Exclusion by the Tunisian law of persons under age, pregnant or breast-feeding women from medical experimentation in the aim of protecting them against clinical research adverse events or abuses is ethically questionable since it deprives them from a possible medical progress. So why not to involve them in clinical research when there is an expected benefit, after bringing them protection as vulnerable persons like we should do for instance for the elderly, handicapped persons or prisoners. Legal creation of research ethics committees is important for the respect of experimentation rules on human beings. PMID- 15822466 TI - [Erysipelas. Retrospective study of 647 patients]. AB - We conducted a retrospective at the department of dermatology of Charles Nicolle hospital of Tunis between January 1994 and December 2000 to determine the epidemiological, clinical profile and the evolution of erysipelas. A total of 647 patients were studied. The mean age was 44.73 years and sex ratio about 1.55. Erysipelas predominately involved in the lower limbs (91.2%). Antecedents of erysipelas were found in 26.12 %. Portal of entry was found in 76.66% represented essentially by toe-web intertrigo. 26.6% of patients were hospitalised. Erysipelas can be controlled with antibiotics; treatment is essentially based on penicillin G 4 mega units intramuscularly every day (60.58%) for mean duration of 10.13 days. Satisfying results were observed in 87.78%. Erysipelas is common disease source of over-morbidity. Many predisposing factors were incriminated, account for the frequency of recurrence, justifying implement of primary and secondary prevention. PMID- 15822467 TI - [Isolated acute renal failure is not associated to an increase in mortality of patients in intensive care units]. AB - This study assess the prognostic factors increasing mortality of patients with acute renal failure in intensive care units. The study included 923 patients admitted to the intensive care unit over a period of two years. Acute renal failure was defined by the following criteria, a blood urea concentration of more than 30 mmol/l, a serum creatinine concentration of more than 180 micromol/l or creatinine clearance less than 50 ml/min. Patients were divided into two groups survivors (V) and dead patients (D). 95 patients presented acute renal failure with a mortality rate of 57.9%. There was no difference between the two groups. There was no difference between the two groups. Patients in intensive care units with only acute renal failure doesn't have a worse prognosis, but the association to two or more other organ dysfunction's, mechanical ventilation of more that 12 days, sepsis or Glasgow coma scale less than 8 are predictive of mortality. PMID- 15822468 TI - [High blood presure for people aged more than 60 years in the distrct of Sousse]. AB - High blood pressure (HBP) is one of the most important cardiovascular risk factors all over the world. We conducted a prospective study including 600 elderly people aged more than 60 years in the district of Sousse. The mean age of our patients was 69 +/- 7.1 years with a ratio of 1.3 (340 females and 260 males). The prevalence of HBP was at 69.3%, diabetes 23% and obesity 24.2%. It is important to note that in 31% of cases HBP was newly diagnosed during our investigation. In 51.7% of patients with known hypertension there was evidence of non compliance to treatment and uncontrolled PB. It is obvious that HBP is a major health problem in our country and that inadequate control of hypertension is a potential source for cardiovascular problems. Our findings emphasize the need for implementing an educational program as well as sensitization compaigns targeting people, particularly those aged more than 60 years. PMID- 15822469 TI - [Germs that produce the extended spectrum betalactamases]. AB - This retrospective study analyses an epidemic with germs ESBL that supervenes at the department of hematology and pediatric oncology in UHC Ibn Rochd of Casablanca. The responsible germ is the ESBL Escherichia coli. Six patients have been infected during the same period that 2 are female and 4 are male. Five patients had acute lenkemia, one patient had a non Hodgkin's disease. All the patients were in the stage of a deep postchermotherapy neutropenia. The picture of all the patients represented a severe infection with suffered fever and acute diarrhea. Five patients died with apicture of septic shock in the 48 to 72 hours after the beginning of the infection and before the identification of the germ. Their treatment consisted in the third generation of cephalosporin and aminoside. One patient who use the imipeneme more the aminoside has been apyrexized the epidemic and severe situation led to the closing of the unit during a week in order to do a disinfection. After 12 monthes of recession, few isolate episodes of infections with enterobacteries ESBL have observed and controlled. The factors that determine the increase and the diffusion of the ESBL germ are numerous and some of them are still not identified, the means of prevention consisted in: the fight against the selection of the resistant germs, the fight against the colonization of the patients by these germs and their transmission between the patients, this requires measures of hygiene and particularly the washing of the hands. PMID- 15822470 TI - [Children bronchiolitis: multicentric study in Tunis]. AB - This study is made in the general framework of the basic health cares program set up by Tunis regional service to fight child serious respiratory infections. It has an objective to measure bronchiolitis winter frequency for patients aged from 1 to 36 months, serious cases frequency, changing profile, the therapeutic behavior and environmental risk factors. This is a multicentric transversal study in grappa, it has taken place in Tunis between January and march 1999 with a sample size of 1340 patients. Result of the survey has shown a medium frequency of the disease with 10% of the patients, median age group infected at 9 months, aggravating factor in 73% of cases, and a surinfection in 70% of cases. Promiscuity is the only environmental risk factor which is statistically significant. Hospitalization was necessary in 14.2% of cases. With regard to prescriptions, bronchodilators, corticoids and antibiotics were prescribed for respectively 71, 39, and 45.6% of patients. Basic health cares structure is a good alternative during the winter season, provided human expertise's are constantly updated and adequate technical staff is available. PMID- 15822471 TI - [Epidemiologic and cytologic configuration of acute leukaemia at University Hospital of Brazzaville]. AB - The authors report the epidemiologic and cytological aspects of 77 patients hospitalized between January 1995 and December 2002 in the clinical hematologic service in University hospital complex of Brazzaville, greatest hospital of the country. During this period, 7155 patients were hospitalized in this service as a frequency of (107%). reported to the number of inpatients admitted on a total of 52,458 patients. The hospital frequency of disease is 0.15%. There's no specific age for this affection (age median = 21-24 years) and all socio-professional categories can be affected with a prevalence of low income patients what complicates extremely the treatment Ratio Man/Woman is 0.8 showing a light female prevalence. On the level of the epidemiologic investigation (limited because of the insufficiency of the means), nothing is retained except rare acute leukaemias secondary to chronic myeloproliferative disorders as well as association with 2 cases of homozygous sickle cell anaemia. Mortality by acute leukaemia in the service is very high, due to lack of equipment. In the cytological plan, there's a light prevalence of the cases of acute lymphoblastic leukaemias of which some could profit from an immunological typing compared to the cases of myeloblastic acute leukaemias PMID- 15822472 TI - [Vitamin A effects on morphometric adaptation after extensive small bowel resection in the rat]. AB - This work was conceived to study, in the rat short bowel syndrome, the effect of precocious administration of retinoic acid on morphometric adaptation in the remnant intestine. Mitotic index, villous height, crypt depth and mucosal aspect were determined in 2 groups of rats (n=12) after 60% small bowel resection. 100 microg of retinoic acid were perfused immediately after resection in the first group. Control group rats received placebo. Two similar groups (n=12) underwent intestinal transection with or without retinoic acid perfusion. Our results show that retinoic acid induced precocious and significant enhancement of all morphometric parameters when compared to placebo. PMID- 15822473 TI - [Congenital elevation of the scapula]. AB - 21 cases of Sprengel deformity in 19 patients were treated in Paediatric orthopaedic unit of Children's hospital of Rabat in 20 years. Only 6 patients (4 girls, 2 boys) were operated on. The ages of the patients at operation ranged from four to fifteen years (mean age: 10 years). The duration of follow-up ranged from 8 months to 10 years (average: 4 years and 2 months) The Woodward procedure was constantly used. Post operatively, the average increase in total abduction was 40 degrees (range 30 to 75 degrees) Functional and cosmetic results were judged good in 3 patients, moderate in 2 and fair in one case. We emphasize that patients who are less than 4 years old are the ideal candidates for scapular displacement procedure. PMID- 15822475 TI - [Retroperitoneal paraganglioma: case report]. AB - Paraganglioma are rare neoplasms arising from undifferentiated cells of the primitive neural crest. We report a case of a 53 years old patient presented with a large tissular retroperitoneal tumour situated above the left kidney. Serum Adrenaline, Nor drenaline and VMA levels were normal. Surgical complete resection of the tumour was performed without intra-operative incident. The histopathological examination and the immunohistochemical analyses concluded to a non secreting retroperitoneal paraganglioma. Postoperative course was uneventful. PMID- 15822474 TI - [Two cases of cancerization of esophageal stenosis due to a caustic burn]. AB - The incidence of carcinoma of the esophagus within patients with chronic esophageal stricture caused by ingestion of corrosive agents is reported to be significantly higher than in the general population. Two patients developped carcinoma of the esophagus respectively 25 and 40 years after corrosive injury. One of these patients had initially gastrostomy and repeated esophagal dilation. Taking into account the high incidence of carcinoma in the site of esophageal stricture, we conclude that the resection of the esophagus is indicated in patients with chronic caustic stricture if there is any finding suggestive of malignancies such as a long duration of the lesions more than 20 years particulary when the ingested agent was caustic soda or sudden aggravation of preexisting dysphagia. PMID- 15822476 TI - [Jaccoud arthropathy and pyrophosphate arthropathy: a rare association]. AB - Jaccoud's arthropathy (JA) is a chronic deformity affecting hands and feet, which are voluntarily correctable by the patients. JA was usually reported in association with rheumatic fever and systemic lupus erythematosus. We describe an exceptional association between a pyrophosphate arthropathy and JA of the hands. It is a 48-year-old woman, having a 13 years history of polyarthritis involving shoulders, elbows, hips and knees, and 4 years after, a progressive unlar deviation of the fingers of the hands. Calcium pyrophosphate dihydrate crystals were identified in the synovial knee biopsy. Hands x-rays as RMI don't identify erosions and confirmed the JA. PMID- 15822477 TI - [Chronic-contained ruptured aortic aneurysm: un unusual cause of back pain]. AB - Chronic contained rupture of abdominal aortic aneurysm is a rare event which can cause diagnostic difficulties. It can present as a chronic back pain and the delayed diagnosis and delayed surgical repair may compromise the final results. The outcome of urgent repair of a chronic contained leak is equivalent to that of elective aneurysm repair. We report a case of contained rupture of a small abdominal aortic aneurysm with delayed diagnosis, evaluated by computed tomography showing a beginning erosion of the lumbar vertebral body. The patient was operated on within 24 hours on admission with uneventful surgical outcome. PMID- 15822478 TI - Implant placement in developing patients. PMID- 15822479 TI - Restorative space management: treatment planning and clinical considerations for insufficient space. AB - In attempting to provide a restorative solution for cases that have been compromised by spatial considerations, clinicians have traditionally opted for an orthodontic approach that did not provide optimal aesthetics due to changes in tooth morphology, specifically tooth size and shape as a result of dental deterioration. With the advent of contemporary aesthetic materials and preparation techniques, clinicians and technicians are now empowered to deliver a penultimate result with minimal compromise to the surrounding dentition. This article presents the clinical and laboratory considerations that must be addressed when providing a prosthetic restoration for crowded teeth. PMID- 15822480 TI - Selecting a luting cement: part II. PMID- 15822481 TI - A delayed subopaquing technique for treatment of stained dentition: clinical protocol. AB - Teeth with severe tetracycline staining can present a restorative challenge to the clinician. While a variety of options facilitate predictable aesthetic treatment of patients with such dentition, they often require significant tooth reduction in order to provide the dentist and the laboratory technician with adequate space for the restorative material to conceal the discoloration. This presentation outlines an alternative to this treatment that uses resin cement to mask the discolorations prior to the cementation of the porcelain laminate veneers. PMID- 15822482 TI - Accuracy of fit and microleakage in ceramic restorations. PMID- 15822483 TI - Achieving anterior aesthetics in the geriatric patient using full-coverage metal ceramic crowns. PMID- 15822484 TI - Scanning and presenting slides: part II. PMID- 15822485 TI - Lasers in modern caries management--part II: CAMBRA. AB - Part two of this series discussed the key strategies that each practice should focus on for caries management. History has proven that oral hygiene and "drilling and filling" alone will not eliminate dental caries. Chemical treatments to prevent and reverse early lesions and conservative, tooth preserving restorative procedures when surgical intervention is necessary should be the new standard of care. Caries management by risk assessment (CAMBRA), where risk factors are "re-balanced" to that of health, is a sound strategy that is one step closer to "curative" dentistry and improving the quality of life of dental patients. The final article in this series will discuss the role that glass ionomer materials and hard tissue lasers play in the minimally invasive restorative procedures for dental caries. PMID- 15822486 TI - Fabrication of an implant-supported reconstruction utilizing CAD/CAM technology. AB - Recent scientific and technological developments have allowed the dental team to execute revolutionary restorative treatments that would have once been considered unfeasible. This article describes a restorative alternative for fixed implant supported reconstructions, consisting of a CAD/CAM-generated framework and CAD/CAM-generated all-ceramic cement-retained restorations. Distinct advantages of this type of restoration include the passive fit of the framework, the ability of the ceramist to fabricate individual restorations independent from the framework, and the delivery of optimal aesthetics. PMID- 15822488 TI - Total arterial myocardial revascularization: analysis of initial experience. AB - BACKGROUND: Total arterial myocardial revascularization is an attempt to overcome the problems of late vein graft atherosclerosis, occlusion and need for coronary re-operations. Despite increasing evidence of efficacy, the use of arterial conduits has not been accepted as a primary practice in most of the centers in Pakistan for various reasons. We analyze our initial experience to assess the feasibility of total arterial revascularization as a primary strategy in patients requiring first time coronary artery bypass grafting. METHODS: Two hundred patients undergoing first time CABG at our institution, from January 2000 to April 2001, were studied. Group 1 consisted of 100 patients undergoing total arterial revascularization (using bilateral internal thoracic and radial arteries) and Group 2 consisted of 100 patients undergoing conventional CABG (using one internal thoracic artery and supplemental veins). Thirty-day mortality and early morbidity with particular reference to resternotomy for bleeding, cerebrovascular accidents, renal failure, and sternal dehiscence were the main outcome measures. RESULTS: Patients in Group 1 were younger (56.2 +/- 10.4 vs. 60.3 +/- 9.8 years; P < 0.001), had lower Parsonnet scores (4.8 +/- 0.4 vs. 9.6 +/- 1.8; P < 0.001), and had better left ventricular function. Both groups received a similar number of grafts. The percentage of patients undergoing total arterial revascularization rose from 20% in the first three months to over 65% in the three later three-month periods. Overall 30-day mortality was 1.5%, one patient (1%) in Group 1 and two patients (2%) in Group 2. There was a similar incidence of postoperative complications and length of median postoperative stay in both groups. CONCLUSION: Total arterial revascularization can be adopted as a primary strategy in most patients undergoing CABG with no increase in mortality or morbidity. PMID- 15822487 TI - Lung cancer. AB - Lung cancer is the leading cause of cancer-related mortality. Since tobacco smoking is the cause in vast majority of cases, the incidence of lung cancer is expected to rise in those countries with high or rising incidence of tobacco smoking. Even though populations at risk of developing lung cancer are easily identified, mass screening for lung cancer is not supported by currently available evidence. In the case of non-small cell lung cancer, a cure may be possible with surgical resection followed by post-operative chemotherapy in those diagnosed at an early stage. A small minority of patients who present with locally advanced disease may also benefit from pre-operative chemotherapy and/or radiation therapy to down stage the tumor to render it potentially operable. In a vast majority of patients, however, lung cancer presents at an advanced stage and a cure is not possible with currently available therapeutic strategies. Similarly, small cell lung cancer confined to one hemi-thorax may be curable with a combination of chemotherapy and thoracic irradiation followed by prophylactic cranial irradiation, if complete remission is achieved at the primary site. Small cell lung cancer that is spread beyond the confines of one hemi-thorax is, however, considered incurable. In this era of molecular targeted therapies, new agents are constantly undergoing pre-clinical and clinical testing with the aim of targeting the molecular pathways thought be involved in etiology and pathogenesis of lung cancer. PMID- 15822489 TI - Pattern of childhood blindness at a referral center in Saudi Arabia. AB - BACKGROUND: An understanding of the causes of blindness and the magnitude of the problem is crucial in designing effective intervention and prevention programs. We undertook this retrospective review to determine the causes of childhood blindness at an eye referral center. PATIENTS AND METHODS: We reviewed charts of children who presented between August 1997 and August 2003. All children had a complete ophthalmologic examination. Blindness was defined as a visual acuity < 20/400, visual impairment as visual acuity between 20/400 and 20/60, and visual loss as a visual acuity < 20/60. RESULTS: A total of 5217 children included 220 (59%) males and 152 (41%) females (age range 2 to 18 years, mean age, 10 years). One hundred twelve (2%) were blind and 260 (5%) had visual impairment. The most common causes of bilateral blindness included optic nerve diseases, retinal disorders, and cataract. The most common causes of unilateral blindness included trauma, retinal diseases, refractive errors, and optic nerve diseases. The most common causes of bilateral visual impairment included refractive errors, corneal diseases, retinal disorders, cataract, and congenital nystagmus. Genetically determined disorders were observed in 37 (70%) of 53 patients with bilateral blindness, and in 52 (56%) of 93 patients with bilateral visual impairment. The incidence of consanguinity among parents of children with acquired causes was 2 (3%) of 59 patients compared to 34 (38%) of 89 among genetically determined causes (P < 0.001). CONCLUSION: Genetically determined disorders continue to play an important role in the causation of childhood blindness among patients attending our referral center in Saudi Arabia. Genetic counseling, early eye screening of children and public education may help in the prevention of visual disorders in children. PMID- 15822490 TI - Antibiotic prescribing patterns for sore throat infections in a university-based primary care clinic. AB - BACKGROUND: Recent studies reveal that a high percentage (over 50%) of episodes for upper respiratory tract infections (URTIs) are treated with antibiotics, regardless of appropriateness or the necessity for prescription. We identified antibiotic prescriptions in a primary health care centre (PHC) and evaluated their suitability for sore throat infections. We also explored whether symptoms, signs, diagnosis and antibiotics prescribed differed by gender. PATIENTS AND METHODS: We collected data on all patients visiting the centre over a period of 12 weeks with a main complaint of sore throat who were prescribed antibiotics after taking a blood count and throat culture. Patients older than 16 years of age were included in the study irrespective of sex, nationality, marital status, occupation or location of residence. The chi square (chi2) statistical test was used in comparing categorical variables. A P value of < 0.05 was considered significant. RESULTS: During the period of study, 579 patients with URTIs presented to the health centre, from which 339 patients with a sore throat were enrolled. Of the study group, 48.7% (165) were male and 51.3% (174) female, with the majority of patients being under 30 years old (54.3%). Throat cultures were positive in 56 patients (16.5%). Most of patients were diagnosed as having pharyngitis (22.7%), and the most frequently prescribed medicine was an oral penicillin (39.1%). Two hundred eight-six patients (84.4%) had 2 or fewer Centor criteria. CONCLUSIONS: Throat cultures were positive in only 16.5% of the patients prescribed antibiotics. This indicates that physicians in the health centre of the university are prescribing antibiotics inappropriately and inconsistently. This also highlights the need for more prescriber education, especially as the range of medications available to the general practitioner for prescribing increases. PMID- 15822492 TI - Neurohormonal changes in medical students during academic stress. AB - BACKGROUND: Academic stress is a good model of psychological stress in humans and is thus useful for studying psychoneurohormonal changes. The aim of the current study was to examine the effect of academic examination stress on activation of the hypothalamus-autonomic nervous system (HANS) and the hypothalamic-pituitary adrenocortical (HPA) axis, through the measurements of changes in neuro-hormones during final exams as compared to the pre-exam baseline. MATERIALS AND METHODS: Forty-eight first- and second-year female medical students participated. Plasma leptin, neuropeptide Y (NPY), nitrite, nitrate, andrenomedullin, cortisol and adrenocorticotropic hormone (ACTH) were measured at baseline and during final examinations. RESULTS: Plasma levels of cortisol, ACTH, NPY, adrenomedullin, nitrite and nitrate increased during times of academic stress as compared to baseline levels. However, only plasma leptin level was decreased during the academic stress as compared to baseline, probably through a negative feedback mechanism resulting from sympathetic stimulation. The results indicate that both the HANS and HPA are involved in this type of stress and both are activated at the same time. CONCLUSION: Academic stress induced significant neurohormonal changes. Leptin, NPY, nitrite, nitrate, adrenomedullin, cortisol and ACTH can be considered part of a complex mosaic model of the neuroendocrine system during academic stress. PMID- 15822491 TI - Primary hyperparathyroidism in north India: a description of 52 cases. AB - BACKGROUND: Primary hyperparathyroidism (PHPT) has a variable clinical expression. Symptomatic PHPT is still the predominant form of the disease in many parts of the world, especially developing countries. Because the clinical profile of the disease has changed from that described in the past, we sought to improve our understanding of the disease in patients in north India. METHODS: We summarized the clinical presentation, biochemical and radiological features, and operative findings from the case records from the last 13 years of 52 patients at a tertiary care centre in north India who had documented PHPT. RESULTS: The male: female ratio was 1: 3.3 with ages ranging from 6 to 60 years (mean +/- SD, 36.38 +/- 12.73). Bone disease (46%), recurrent renal stones (21%) and body aches and pains (21%) were the most common modes of presentation. The lag time varied ranged from 1 month to 16 years. Common clinical manifestations included bone pain (67%), weakness/fatigue (56%), fracture of the long bones (48%), abdominal pain (39%), polyuria (37%) and psychiatric manifestations (23.1%). Hypertension was observed in 42% and a palpable nodule in the neck in 19%. Biochemical features included hypercalcemia (86.5%), hypophosphatemia (65.4%) and hyperphosphatasia (67.3%). Mean intact PTH ( +/- SD) was 809.0 +/- 696.3 ng/L with levels significantly lower in patients who had only kidney stone disease as compared with those with bone disease (P = 0.017). A single parathyroid adenoma was localized in 50 (98%) patients. Hungry bone disease was seen in 59% patients. CONCLUSION: PHPT in India continues to be a symptomatic disorder with skeletal and renal manifestations at a much younger age. PMID- 15822493 TI - Morphometric examination of the paranasal sinuses and mastoid air cells using computed tomography. AB - BACKGROUND: The paranasal sinuses and mastoid air cells vary considerably in size and shape from person to person. The main structures are pneumatic. In this study, we investigated the relationship between right and left sides and evaluated the volume changes according to age and sex. METHODS: Of all patients attending the radiology department, 91 cases without paranasal sinuses and mastoid air cells pathology (i.e., inflammation, operation or trauma) were selected for evaluation. Axial computed tomography (CT) scans were obtained for both paranasal sinuses and temporal bones. In all scans, the volumes of each area (maxillary sinus, frontal sinus, sphenoid sinus and mastoid air cell) were calculated and analyzed statistically. RESULTS: The volumes of paranasal sinuses and mastoid air cells increased with age and women had a lower mean volume. There was a positive correlation between right-left and ipsilateral structures (paranasal sinuses and mastoid air cells). CONCLUSIONS: These results are helpful in understanding the normal and pathological conditions of the paranasal sinuses and the mastoid air cells. PMID- 15822494 TI - Documentation and coding of medical records in a tertiary care center: a pilot study. AB - BACKGROUND: Since the medical record is the major source of health information, it is necessary to maintain accurate, comprehensive and properly coded patient data. We reviewed 300 medical records from patients at King Faisal Specialist Hospital and Research Center, representing four departments (medicine, surgery, pediatrics and obstetrics and gynecology). METHODS: The records were audited following the guidelines of the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) for accuracy and completeness of documentation and coding of primary and secondary diagnoses and procedures performed. RESULTS: Of 1051 items abstracted, 876 (83.3%) were accurately documented, 41 (3.9%) were inaccurately documented, and 134 (12.7%) were not documented. Of the items abstracted, 736 (70%) were assigned a correct code, 110 (10.5%) were assigned an incorrect code, and 205 (19.5%) were not coded. More items classified as accurately documented were coded correctly (71.1%) than items inaccurately documented (49.7%) (P < 0.0001). The difference in comprehensiveness of documentation, which reflects physician performance, was not statistically significant among the four departments (P value < 0.234). The difference in the accuracy of coding, which reflects coder performance, was statistically significant (P value < 0.036). CONCLUSIONS: Only 60% of the audited records met the benchmark for good quality medical records with regards to documentation and coding. A positive correlation between the accurate documentation and correct coding was noted, which supports the conclusion that high quality documentation enhances coding accuracy. These data, although encouraging, suggest room for improvement, which can be achieved through the collaboration of clinicians, who have extensive clinical experience, and coding professionals, who have comprehensive classification system expertise. PMID- 15822495 TI - Malakoplakia associated with colorectal adenocarcinoma. PMID- 15822496 TI - Acute chest syndrome in adult sickle cell disease in eastern Saudi Arabia. PMID- 15822497 TI - Concealed penis in a 2-year-old boy: a rare complication of circumcision. PMID- 15822498 TI - Spontaneous perforation of the common bile duct in an adult. PMID- 15822499 TI - Congenital cystic adenomatoid malformation associated with esophageal duplication cyst. PMID- 15822500 TI - Geographical distribution of congenital heart defects in Saudi Arabia. AB - BACKGROUND: Congenital heart defects (CHD), which are caused by abnormalities early in fetal life, encompass over 50 diagnoses. Since the detailed etiology is unknown, the geographical distribution of defects might suggest likely risk factors. METHODS: The geographical distribution of 5 865 Saudi Arabian nationals with CHD was studied by cross-matching their residential provinces and towns with a geographical information system provided by the General Directorate for Military Survey. Population data were obtained from the 1413H census. RESULTS: CHD cases were mostly distributed across the provinces in proportion to their total population but due to their size and inhomogeneity, province-based thematic maps were found to be misleading. City-based maps were preferable and showed similar geographic distributions for cases registered in successive years. Thematic maps of the distribution of the CHD burden highlighted the southwestern provinces, nearthe border with Yemen, and the northeast section of the Eastern Province. CONCLUSIONS: Patterns of disease in Saudi Arabia are best studied at the level of individual towns and villages. The CHD registry has already attained good national coverage and can therefore support nationwide epidemiological studies. Southwestern Saudi Arabia and the northern part of the Eastern Province appear to exhibit a higher burden of CHD. PMID- 15822501 TI - The radial artery as a coronary bypass conduit: dealing with hypereactivity. PMID- 15822502 TI - A case of previously undiagnosed Crohn's disease presenting with acute pancreatitis as an extraintestinal manifestation. PMID- 15822503 TI - A new model for child survival in Africa. PMID- 15822504 TI - Eliminating iodine deficiency worldwide is within reach. PMID- 15822505 TI - [Indications and results of infliximab in Crohn's disease]. AB - Infliximab is a chimeric monoclonal antibody to tumor necrosis factor a. Induction therapy with infliximab is indicated for treatment of refractory Crohn's disease and for reduction in the number of draining fistulas in fistulizing Crohn's disease. Indications for maintenance therapy with infliximab include maintenance of remission in active Crohn's disease who responded to initial induction therapy with infliximab and maintenance of fistula improvement who responded to initial therapy with infliximab. Side effects of infliximab are infusion reactions, immunogenicity, induction of DNA antibodies and infection complications including tuberculosis. PMID- 15822506 TI - [Reactive arthritis]. AB - Reactive arthritis are definied as steriles arthropathies using classic bacteriological techniques. They are due to extra articular infection and are often associated with HLA B27. The outcome of these arthritis is characterised by the recurrence of flares with sometimes appearition of ankylosing spondylitis. The pathogenesis of reactive arthritis is modified when bacterial antigens or alive micro-organisms are discovered in involved joints. Several current works have underlined the interest of antibiotic therapy in the chlamydial reactive arthritis. Chronic forms can justify the use of anti-rheumatic drugs such as salazopyrine. PMID- 15822507 TI - [Pseudomonas aeruginosa: a multicentric study of antibiotic resistance (1999 2000)]. AB - A retrospective multicentric study was carried out over a period of 2 years (1999 2000). 2659 strains of Pseudomonas aeruginosa were collected from 4 university hospitals (Charles Nicolle Hospital, Pediatric Hospital and National Centre of Bone Marrow Transplantation in Tunis, Habib Bourguiba Hospital in Sfax). Epidemiological profile and antibiotic susceptibility were analysed. All bacteria were identified by conventional methods and antibiotic susceptibility tests were performed according to CA-SFM guidelines. The strains were recovered essentially from surgical wards (33%) and intensive care units (22%). Pseudomonas aeruginosa was isolated mainly from pus (36%), urine (32%) and respiratory samples (18%). 25% of strains were resistant to ticarcilline, 18% to cefsulodine, 9% to ceftazidime, 14% to imipenem and amikacin and 25% to ciprofloxacin. Moreover, the resistance rates varied from hospital to hospital and from unit to another. The resistant strains were isolated particularly from urology and intensive care units: respectively 62% and 39% for ticarcilline; 26% and 13% for ceftazidime. The acquired resistance to b-lactams seems largely due to penicillinase production. The frequency of resistance to ceftazidime was the lowest and seems associated to chromosomal cephalosporinase over production. PMID- 15822508 TI - [Uterine fibromyomas: fertility after myomectomy. About 41 cases]. AB - Fibromyomas are among the most frequent pathologies of women in their reproductive years; that's why clinicians studied their impact on fertility especially when no other cause is found. In this retrospective study of 41 infertile patients between January 1996 and December 1999, the authors tried to define the assess of myomectomy in these cases and the relevant factors that influence the post-operative results in terms of fertility. 24 patients were operated by laparotomy, 7 by laparoscopy and 10 by hysteroscopy. Once surgery was performed, 19 women conceived (46.34%) with delays ranging between 3 and 36 months (mean delay: 15.5 months). An age of less than 40, a sterility of less than 5 years duration and the absence of associated factors seems to enhance the chances of these patients to conceive after mytomectomy. No significant difference was noted in terms of subsequent fertility between the group of primary infertility and that of secondary sterility; neither concerning the myomas' number, size or place. Pregnancy rates after surgery were equivalent whether the myomectomy was performed by laparotomy or laparoscopy. It is, then, reasonable to propose to infertile patients with uterine myomas, a conservative surgery especially if they are young and if their sterility is unexplained for less than five years. PMID- 15822509 TI - [Cardiac involvement in spondylarthropathies]. AB - Cardiac involvement is a real manifestation of spondylarthropathies and include specially aortic regurgitation and conduction troubles. We present a prospective open study of fifty patients with spondylarthropathy (responded to Amor criteria) in order to evaluate the frequency of cardiac involvement, to see its type and if we can evaluate a group of patients able to this complication. We have included forty-four men and six women with a mean age of 38 years. The disease evolve for 8.8 years in mean. Ag HLA B27 was present in 70% of the cases. Thirty patients have ankylosing spondylitis, although twenty have a secondary spondylarthropathy: psoriasic rheumatism (12 cases), inflammatory bowel disease: Crohn's disease (4 cases), RCH (three cases) and Fiessenger le Roy-Reiter syndrome in one case. All the patients have had a cardiac check up with research of clinical cardiac manifestation, thoracic chest, trans-thoracic echographi, Halter rhythmic done in five cases only. Cardiac involvement is found in five cases (10%): aortic regurgitation in 3 cases (6%) and mitral regurgitation in 2 cases (4%). These valvular disease are well tolerated. PMID- 15822510 TI - [First seizure in children. Exploration strategy]. AB - We have carried out a retrospective study on 51 children aged between 2 years 3 months and 13 years in order to determine the findings and to define the indications of neuroimaging modalities in children with a first unprovoked seizure. Children who had neuroimaging studies were divided into two groups based on the results of neuroimaging normal or abnormal. We have compared the two groups according to each clinical and electroencephalographic parameters studied. Neuroimaging abnormalities were found in 47% of cases. According to our results and literature review, we concluded that the imaging should be done if we have at least one of the criteria: an abnormal neurological examination after the seizure and focal slowing waves on the electroencephalogram. PMID- 15822511 TI - [Chemo-laparoscopic treatment of hydatic cyst of the liver myth or reality! Preliminary results of 27 cases]. AB - The laparoscopic treatment of hydatic cyst of the liver is at present codified. These last years, several progresses were obtained in the laparoscopic approach of hydatic cyst by avoiding the risk of preoperative hydatic peritoneal scattering. The aim of this study is to evaluate the effect of Albendazole given before a laparoscopic management of the hydatic cyst of the liver. PMID- 15822512 TI - [Mucus secreting tumors of the appendix. Report of 22 cases]. AB - The retrospective study presented in this article was realized on a series of 22 cases of mucus secreting tumor of the appendix compiled among 8373 appendicectomies. The clinical picture was dominated by an appendiceal syndrome (73%). The diagnosis was made pre or peroperatively in 10 cases and it was only with to the systematic anatomopathologic exam that 12 other cases were discovered. The histological study of the appendectomy piece showed a retention mucocele (12 cases), a mucosal hyperplasia (8 cases) and a cystadenoma (2 cases). For 5 patients, the diagnosis of peritoneal pseudomyxoma (4 cases) or extraperitoneal (1 case) was retained. All the patients underwent an appendicectomy, associated to an evacuation of the peritoneal gelatinous ascites for 5 patients among whom two underwent in more an omentectomie. Two cases of recurrence were recorded 5 months later. PMID- 15822513 TI - [The immediate effect of Rashkind's atrioseptostomy on systemic saturation in transposition of the great arteries]. AB - The purpose of this prospective study is to analyse the immediate effect of Rashkind's atrioseptostomy on systemic saturation in transposition of the great arteries (TGA). Thirteen neonates and infants (10 males and 3 females) with TCiA underwent balloon atrial septostomy (BAS) at a median age of 20 days (range 2 and 60 days). The mean atrial septal defect diameter after BAS was 6.5 +/- 1.1 mm. The right ventricular saturation increased from 37 +/- 17% to 67 +/- 13% (p < 0.001). There was no correlation between the atrial septal defect diameter and the increase of systemic saturation after BAS. PMID- 15822514 TI - [Sclerosing stromal tumor of the ovary. Report of 3 cases]. AB - Sclerosing Stromal Tumor (SST) is an uncommon benign tumor of the ovary, distinct from thecoma and fibroma because of predominant occurrence below 30 years of age, lack of hormonal manifestations and histologic heterogenity. Three cases of SST occuring in a 15, 26 and 56-year-old females are reported. Review of the literature and clinicopathological features are described in the present article. Histogenesis and differential diagnosis are also discussed. PMID- 15822515 TI - [Peripartum pubic symphysis separation. A report of three cases and review of the literature]. AB - Peripartum pubic symphysis separation is a rare complication of delivery, but the incidence seems to be under-estimated. We report three cases of pubic syphysis separation identified in a two-year period. All cases occurred following spontaneous non operative vaginal deliveries. The underlying etiology and pathophysiology has not been fully elucidated. Thus, prevention is difficult. Conservative therapy usually results in a complete recovery within many weeks. PMID- 15822516 TI - [Acral acanthosis nigricans paraneoplastic: about one case]. AB - The paraneoplastic acanthosis nigricans occurs in association with visceral neoplasms. We report a case of acanthosis nigricans that seat at a dorsal face of the second, third and fourth right toe of a 63 year-old patient. The exploration has been put in evidence a carcinoma of the right lung. The acanthosis nigricans has been nearly disappeared after resection of the pulmonary lobe. The relapses of the acanthosis nigricans in a same sit after five years has been reveal a carcinoma of the bladder. The resection of the bladder tumour has been followed by disappearance of the acanthosis nigricans. PMID- 15822517 TI - [Radiation-induced intracerebral cavernous angioma]. AB - Recently, there have been several case reports of cavernous angiomas appearing after radiation to the brain, typically in children. The time interval between irradiation and the detection of the cavernous angioma varied from three to nine years and the doses from 24 to 60 grays. We report the appearance of a cavernous malformation in a young girl 3 years after radiation therapy for a brain tumor. PMID- 15822519 TI - Fast track to rewards. In the pay-for-performance event, everyone wins. PMID- 15822518 TI - [Gestationis pemphigoid. From three observations and in the light of a review of the literature]. AB - The gestationis pemphigoid is a rare auto-immune vesiculo-bulleuse dermatosis occurring during the pregnancy or of the postpartum. The diagnosis is evoked by the localization and the type of eruption like by which has occurred of successive pushes. The cutaneous biopsy in immunofluorescence confirms the diagnosis by showing the linear deposit of C3 along the basal membrane. The risk of complications fetales is high in the event of pemphigoid gestationis of or the interest of an early diagnosis and a treatment, the ci rests on the corticotherapy in topic or according to the gravity of the symptoms in general administration. From three observations and in the light of a review of the literature, the authors report the characteristics of this pathology. PMID- 15822520 TI - A labor of love. With telemedicine, retired executive Frank Brady brings information to doctors and help to children. PMID- 15822521 TI - The Health IT Sharing project. Its processes and procedures can be the foundation for any collaborative initiative. PMID- 15822522 TI - Pharmacy benefit management. Co-opetition fosters a national e-prescribing standard. PMID- 15822523 TI - Electronic documentation. Jewish Hospital staff helped implement and now embrace its clinical information system. PMID- 15822524 TI - WORM-enabled tape storage. The method meets security standards and delivers the benefits of tape. PMID- 15822525 TI - Urinalysis interpretation: how to squeeze out the maximum information from a small sample. AB - The urinalysis is an essential part of the diagnostic evaluation for all urinary and many metabolic diseases. Its assessment includes evaluation of physical characteristics (color, clarity, and volume), biochemical parameters (urine pH, blood, glucose, ketones, bilirubin, urobilinogen, and protein) and microscopic sediment evaluation (RBC, WBC, organisms, epithelial cells, crystals, and casts). Many of these parameters are influenced by collection method and therefore, it is essential to interpret accordingly. Knowledge of factors that can interfere with the accuracy of some test results can decrease improper interpretation. When all of these parameters are evaluated in combination with clinical signs, physical examination, thorough history and other laboratory tests, a diagnosis will often be attained. PMID- 15822526 TI - Renal biopsy of dogs and cats. AB - Renal diseases are common in dogs and cats. Renal biopsy may be required during the evaluation of the patient to establish a definitive diagnosis, determine the severity of the lesion and formulate an optimal treatment plan. Renal biopsy specimens can be collected via several methods. Percutaneous techniques are performed with ultrasound guidance in both dogs and cats or blindly in cats. If ultrasound guidance is not available, the keyhole technique can be used in dogs. Biopsy can also be performed using laparoscopy or surgery. While complications can arise with any of these techniques, complications are less frequent when an experienced operator uses proper technique. Renal biopsy specimens must be processed and evaluated appropriately if consistent and accurate diagnoses are to be rendered. The article summarizes patient selection and evaluation, renal biopsy techniques, expected complications of renal biopsy, and appropriate processing and evaluation of the renal biopsy specimen. PMID- 15822527 TI - Hypertension in renal disease: diagnosis and treatment. AB - Hypertension is a common sequela to renal disease in cats and dogs, affecting as many as 61% cats and 93% of dogs, respectively. Undiagnosed and untreated, elevations in blood pressure can have deleterious effects on the brain and heart as well as promote further renal injury. In this article, we discuss the identification of patients at risk for hypertension as well as methods for measuring blood pressure and the treatment of hypertensive patients. PMID- 15822528 TI - Glomerular filtration rate and renal scintigraphy. AB - Clinical evaluation of early renal dysfunction is problematic in the nonazotemic animal. Measurement of glomerular filtration rate (GFR) provides the most accurate assessment of renal function. However, many methods of determining GFR are not practical for use in routine veterinary practice. This article provides a description and literature review of methods to determine GFR, including benefits and drawbacks of each method for the practicing veterinarian. PMID- 15822529 TI - Excretory urography. AB - Excretory urography is a type of contrast study used to verify and localize upper urinary tract disease. In some instances, information regarding renal function and disease pathophysiology can also be obtained. With the recent advances in small animal ultrasonagraphy, excretory urography has become an underutilized procedure. This article will help explain why excretory urography remains, and will remain, a ubiquitous test that gives excellent detail of the entire urinary tract, and remains an essential tool for the assessment of the renal pelves and especially the ureters. Specifically, this article will focus on technique and interpretation of a properly performed excretory urogram. PMID- 15822530 TI - Contrast cystography. AB - Cystography is a radiographic study performed to aid in evaluation of the urinary bladder for extramural, mural, or intraluminal lesions. These lesions may primarily involve the urinary bladder or may be an extension of disease from adjacent organs. Cystography is easy to perform with relatively few complications. Different types of cystography (positive versus negative contrast) may be used depending on the type of information that the clinician hopes to obtain. Although a valuable technique, it is important to correlate the findings on cystography with other clinical information to arrive at the final diagnosis. PMID- 15822531 TI - Cystoscopy: techniques and clinical applications. AB - Cystoscopy is a powerful tool for characterization of lower urinary tract disease in dogs and cats. Current applications of cystoscopy include diagnostic and interventional techniques. This article reviews cystoscopy equipment, procedures, and common applications of cystoscopy. A review of normal anatomy and common lower urinary tract lesions identifiable with cystoscopy is also presented. PMID- 15822532 TI - Urodynamic testing in the diagnosis of small animal micturition disorders. AB - Urodynamic testing provides a quantitative assessment of the function of the small animal lower urinary tract. Most commonly these techniques are utilized to assess urethral tone (urethral pressure profile or UPP) and bladder detrusor muscle function (cystometrogram or CMG). A UPP may be indicated in cases of canine and feline micturition disorders. Examples include suspected primary sphincter mechanism incontinence (PSMI), ureteral ectopia, other congenital abnormalities, suspected neurological disorders, and mechanical or functional urethral obstruction. A UPP can be performed effectively utilizing human dedicated equipment. A CMG may be indicated to assess detrusor function in all of the above cases as well as cases of suspected atonic or infiltrated urinary bladder. This procedure can also be performed using the same human equipment. These tests are useful not only in providing an accurate diagnosis, but also in providing a sensitive prognostic indicator for clinical outcome of micturition disorders with and without pharmacological or surgical therapy. A leak pressure point may also be established in dogs with urinary incontinence, and may be even more sensitive than a UPP to predict clinical incontinence in some cases. PMID- 15822533 TI - Prevalence of thalassemia in pregnant women at Maharaj Nakorn Chiang Mai Hospital. AB - OBJECTIVE: To determine the prevalence of thalassemia including alpha-thalassemia 1 trait (SEA type), beta-thalassemia trait, hemoglobin E (HbE) trait, homozygous HbE, the combination of alpha-thalassemia-1 (SEA type) and beta-thalassemia trait, alpha-thalassemia-1 (SEA type) and hemoglobin E trait, and beta thalassemia hemoglobin E disease in pregnant women. METHOD: A cross-sectional descriptive study was conducted on pregnant women who attended the antenatal clinic at Maharaj Nakorn Chiang Mai Hospital, from 1 August to 31 October 2001. All subjects had blood taken for diagnosis of thalassemia trait or diseases, based on quantitative electrophoresis, and PCR (polymerase chain reaction) technique RESULTS: 516 pregnant women were recruited 81.0% resided in Chiang Mai province, and the remainder were in other northern provinces of Thailand. The mean (+/-SD) age was 27.7+/-6.3 years old. 5.6% of cases had anemia. Overall prevalence of thalassemia trait was 25.4% which were classified as follows: alpha thalassemia-1 (SEA type) trait 6.6%, beta-thalassemia trait 3.7%, hemoglobin E trait 11.6%, homozygous hemoglobin E 0.8%, the combination of alpha-thalassemia-1 (SEA type) and beta-thalassemia trait 1.2% and the combination of alpha thalassemia-1 (SEA type) and hemoglobin E trait 1.5%. Additionally, the authors also found beta-thalassemia hemoglobin/E disease 0.2%. CONCLUSION: The prevalence of thalassemia carriers among pregnant women at Maharaj Nakorn Chiang Mai hospital was high, indicating the necessity of a screening thalassemia program aimed at prevention and control of this disease. PMID- 15822534 TI - Prevalence of bacterial vaginosis in Thai women attending the family planning clinic, Siriraj Hospital. AB - OBJECTIVES: To investigate the prevalence of bacterial vaginosis (BV) among Thai women attending a family planning clinic and to evaluate the association of BV with potential risk factors. MATERIAL AND METHOD: A cross sectional prevalence study was conducted among 800 women attending the family planning clinic, Siriraj Hospital, between August and December 2003. BV was diagnosed according to Amsel's criteria. Prevalence and risk factor models were compiled and statistically analyzed RESULTS: Among the low risk population acquiring sexually transmitted diseases, the prevalence of BV was 14.6% (117 of 800). Asymptomatic disease was recognized in up to 47.9% (56 of 117). BV was significantly more prevalent among those who used douching inside the vagina [OR = 3.98 (1.85-8.33), p < 0.01] and high a prevalence among IUD users [OR = 1.84 (1.22-2.79), p < 0.01]. Although not statistically significant, BV tended to be more prevalent among women with a lower age at first intercourse, higher numbers of lifetime partners, higher frequency of sexual intercourse and current smokers. CONCLUSION: BV is a relatively high prevalent condition. The two potential risk factors, douching inside the vagina and IUD use, can be demonstrated, adding to be more concerned about the inappropriate practice of douching and more consideration in IUD users. The other potential risk factors, the impact to adverse reproductive outcomes and the prevention of BV need further studies, particularly in various Thai populations. PMID- 15822535 TI - Laparoscopic repair of inguinal hernia using two layers of mesh with reinforcement of the inner mesh by 2 sutures. AB - PURPOSE: To determine the effectiveness of the laparoscopic repair of inguinal hernia using two layers of mesh with reinforcement of the inner mesh by 2 sutures. PATIENTS AND METHOD: The hernias included primary, direct, indirect, bilateral inguinal hernias and 5 recurrent inguinal hernias. The present report described laparoscopic repair by the total extraperitoneal mesh placement (TEP) technique using two layers of polypropylene mesh with the inner mesh reinforced by 2 sutures in 20 adult patients with 22 hernias. RESULTS: The operations were successfully performed on all the patients with 1 complication (5.00%) which was treated by surgical procedure. There was no recurrence with a follow-up of 10.27+/-5.96 months (range1.10-20.00 months). CONCLUSIONS: The report has shown that this operation can be used to treat primary, recurrent, direct, indirect, and bilateral inguinal hernias in adults with no recurrence in a short term follow-up. PMID- 15822536 TI - Intertrochanteric fractures of the femur: results of treatment with 95 degrees Condylar Blade Plate. AB - The aim of the present study was to analyze patients treated with 95 degrees Condylar Blade Plate and to describe the technique that can prevent complications. 69 patients with closed intertrochanteric femoral fractures were treated by one surgeon using the 95 degrees Condylar Blade Plate, and it was found that 56 of them met the minimum 10 months follow up requirement. 53 patients (94%) healed after the procedure. There were 4 patients (7%) requiring surgical intervention. One patient required bipolar arthroplasty because of the cutting through of the implant, two patients needed repeated open reduction and internal fixation (ORIF) with bone graft due to delayed union and broken implant, and one patient had an infection which was resolved after debridement and a course of antibiotics. The surgical time averaged 45 minutes and blood loss averaged 150 ml. It appeared that open reduction and internal fixation using the 95 degrees Condylar Blade Plate was effective in treating patients with intertrochanteric femoral fractures. The surgical time and blood loss were minimized. Early patient rehabilitation was initiated, and the complications were decreased. PMID- 15822537 TI - Mammographic findings in breast cancer patients, who were treated with breast conserving therapy. AB - PURPOSE: To evaluate the mammographic findings and role of detection of local recurrence in breast cancer patients, who underwent breast conserving surgery and radiation therapy. DESIGN: Descriptive retrospective study MATERIAL AND METHOD: From 1988 to 1998, 78 women with early breast cancer (stage I, II) were treated with lumpectomy and axillary nodes resection, followed by radiation treatment. Mammographic imagings were taken yearly after complete treatment. Needle biopsy was done for suspected local recurrence on mammographic findings. RESULTS: Duration of follow-up ranged from 3 years to 10 years. Seventy-three of 78 patients (93.6%) had evidence of parenchymal changes on serial annual mammography. Twenty-eight of 73 patients (38.4%) had diffuse dense parenchymal changes. Twenty-four of 73 patients (32.9%) had focal fibrotic changes, and 21 of 73 patients (28.8%) had evidence of parenchymal distortion with a mass like lesion. Skin thickening was found in 54 of 78 patients (69.2%). Calcification was the least change, found in only 2 of 78 patients (2.6%). Nine of 78 patients (11.5%) had mammographic signs of local recurrence. Seven of 9 patients had suspected local recurrence by needle biopsy. Four of seven patients had proven local recurrence on mastectomy, and 3 of 7 patients (42.8%) had false positive on mastectomy. CONCLUSION: Annual mammography in post breast conserving treatment showed beneficial results for detection of recurrent cancer PMID- 15822538 TI - Phytate and fiber content in Thai fruits commonly consumed by diabetic patients. AB - The amount of phytate and total dietary fiber of 6 popular tropical fruits eaten by diabetic patient residing in Bangkok were studied. They were dragon fruit, durian, guava, longan, mango, and pineapple. Fruit containing the highest content of total dietary fiber was guava which had about 2.7%. Longan contained the lowest fiber content equal to 0.19%. The highest content of phytate was found in pineapple about 0.09% and the lowest content was in longan about 0.037%. It was noticed that longan contained the lowest total dietary fiber and the lowest phytate content. PMID- 15822539 TI - A chart audit of postoperative pain assessment and documentation: the first step to implement pain assessment as the fifth vital sign in a University Hospital in Thailand. AB - PURPOSE: To describe the documentation of pain assessment and management in the first 72 hrs postoperatively. DESIGNS: Retrospective descriptive study MATERIAL AND METHOD: Four hundred and twenty five hospital charts in December 2002 were audited to reveal the quality of postoperative pain assessment and documentation. Scores above 21 from the possible maximum of 28 (75%) were accepted for the review. RESULTS: Nurses documented pain assessment more often than doctors (98.8% vs 29.4%). An assessment of pain intensity using a numerical rating scale (0 to 10) was found in 192 (45.2%) charts, and using a pain descriptor scale in 408 (96%) charts. The documentation of pain both before and after giving analgesics was scarce during the first 3 days postoperatively. Apart from charts that used a patient-controlled analgesia (PCA) technique which had a specific record form, regular pain assessment every 2 to 4 hrs during the first 24 hrs was found in only 2 (0.5%) charts. Pain assessment items which were documented inconsistently and below accepted standards were pain assessment after administration of analgesics, pain assessment every 2 hours in the first 24 hours (day 1), and pain assessment thereafter every 4 hours in the first 24-72 hours (days 2 and 3). The other 4 of 7 audit pain items were documented in higher scores: initial patients' pain intensity and sedation assessment, pain treatment, continuity of pain assessment and pain assessor s name scores. Nevertheless, because of the low total audit score [mean+/-SD = 10.7+/-3 out of 28], it was considered that none of the reviewed charts reflected good quality pain assessment and documentation. CONCLUSION: The present study revealed that the existing practices of pain assessment and documentation were poor. The need for development of regular pain assessment as if pain is the fifth vital sign should be widely emphasized as a part of quality assurance. PMID- 15822540 TI - Energy drinks consumption in male construction workers, Chonburi province. AB - This unmatched case-control study aimed to determine the relationship among caffeine drinks consumption known as "energy drinks consumption", drug dependence and related factors in male construction workers in Chonburi Province. It was conducted during December 15, 2001 and February 15, 2002. Data were collected using interview questionnaires. The logistic regression was used to control possible confounding factors. The subjects consisted of 186 cases who had consumed energy drinks for more than 3 months and 186 controls who had given up for more than 3 months. They were frequency/group matched by age group. There was statistically significant association among energy drinks consumption and overtime work, motivation from advertisements, positive attitude of energy drinks consumption, alcohol drinks, smoking and ex-taking Kratom behavior. Multivariate analyses revealed that only 5 factors were related to energy drinks consumption: marital status (OR = 1.88, 95%CI: 1.14, 3.11), overtime work (OR = 2.84, 95%CI: 1.73, 4.64), motivation from advertisements (OR = 2.72, 95%CI: 1.67, 4.42), positive attitude of energy drinks consumption (OR = 4.06, 95%CI: 1.65, 10.01) and ex-taking Kratom behavior (OR = 2.77, 95%CI: 1.19, 6.44). As a result, construction workers should be provided with the knowledge of energy drinks consumption, the effect of drug dependence behavior, and the advantages of safe and healthy food that is cheap, readily available, and rich in nutrients. PMID- 15822541 TI - Quality of life assessment in radiotherapy patients by WHOQOL-BREF-THAI: a feasibility study. AB - OBJECTIVE: To determine the feasibility and sensitivity of the WHOQOL-BREF-THAI assessment tool in measuring the quality of life in cancer patients treated with radiotherapy in routine clinical practice. METHOD: A cross-sectional analytical study including the consecutive radiotherapy patients at Songklanagarind Hospital from October to December 2000 was conducted. The participants completed a Thai version of the brief form of the WHO quality of life assessment instrument (WHOQOL-BREF-THAI). RESULTS: The majority of the patients (80.7%) were able to complete the questionnaire, 19 (12.6%) by themselves, 4 (2.6%) with the help of their relatives and 128 (84.8%) through the interview. Almost all of them could understand the questions very well. The mean and standard deviation of time requirement in completing it were 13.0+/-4.0 minutes. The WHOQOL-BREF-THAI instrument was sensitive enough to discriminate the QOL in patients with differences in all analyzed clinical parameters, which were hospitalization, stage of disease, treatment aim and ECOG performance status. CONCLUSION: The present results supported the feasibility of using the WHOQOL-BREF- THAI assessment tool in radiotherapy cancer patients with good sensitivity and patient understanding within an acceptable time requirement. One practical barrier of concern was a low self-assessment percentage in this particular group of patients, necessitating the interviewer system. PMID- 15822542 TI - Flunarizine for prophylactic treatment of childhood migraine. AB - OBJECTIVE: To determine the effectiveness of flunarizine for migraine prophylaxis in children. PATIENTS AND METHOD: Children aged between 7 and 15 years who had the indication for prophylactic treatment of migraine were recruited into a prospective study at the Department of Pediatrics, Ramathibodi Hospital, from January 1st to December 31st 1999. After verbal consent was obtained, flunarizine was administered either at 5-mg daily in those who had never received it or at 10 mg daily in those who previously took this drug within one year Serial evaluation for the severity of migraine including duration, intensity, and frequency of headache attacks was performed every 2 weeks for 6 months. RESULTS: Twenty-one children (10 boys, 11 girls) with a mean age of 11.3+/-2.48 years (range 7-15 years) were enrolled in the study. There were ten children who had migraine with aura. Initially, 5-mg daily and 10-mg daily of flunarizine were administered in 19 and 2 patients respectively. The dosage was increased to 10-mg daily after two weeks in 5 patients because of the unresponsiveness to the initial dose. Improvement was observed in 14 patients (66%) including 13 of 14 patients who received 5-mg daily and 1 of 7 patients who received 10 mg daily. Five patients (23%) had no recurrent attack. Nine patients (42%) had more than 50%-reduction of frequency of migraine and 3 of these had either shorter duration or less intensity of the attack. Clinical improvement was observed between 2 and 4 weeks after initiation of treatment. There was no adverse effect observed CONCLUSION: This is a preliminary result demonstrating that flunarizine is one of the effective drugs for migraine prophylaxis in children. PMID- 15822543 TI - Evaluation of MR cisternography in diagnosis of cerebrospinal fluid fistula. AB - PURPOSE: To study the clinical usefulness and sensitivity of MR cisternography as noninvasive study in the diagnosis of CSF fistula in patients with clinical diagnosis CSF rhinorrhea or otorrhea. METHOD: Thirty-five patients with clinically diagnosed CSF leakage were examined for site of dural tear with MR cisternography with additional plain high-resolution CT in some cases from Jan. 1999 to Dec.2002. The MR imaging technique was performed as a heavily T2 weighted fast spin echo study with fat suppression in axial, coronal and sagittal projections. Criteria for positive results were demonstrable fistular tract connecting subarachnoid space to paranasal sinus/petrous bone, and/or dural discontinuity, and/or bone defect with pneumocephalus, and/or presence of brain herniation. Eighteen of the patients subsequently had exploratory surgery for fistula. Sensitivity analysis of the surgical results was compared with the findings at MR cisternography. RESULT: MR cisternography showed significant correlation with surgical findings with sensitivity of 89%. Additional high resolution CT were complete agreement with site of fistular tract. The pathogenesis of CSF leakage was related to trauma (86%). The commonly found dural/bony defect and location of connecting fislular tract were cribriform plate and ethmoid sinus of 58%, 55% respectively. CONCLUSION: In the presence of clinically diagnosed CSF leakage, the combination of MR cisternography and plain high-resolution CT are highly accurate in locating the site and extent of CSF fistula and should be considered a viable noninvasive alternative to CT cisternography and Tc-99m-DTPA cisternography. PMID- 15822544 TI - Inappropriate weight management among Thai women consuming anorectics prescribed by private clinics in Bangkok. AB - This cross-sectional survey was conducted by a 2-stage sampling design. A total of 250 women, aged > or = 18 years, were recruited from 10 randomly chosen private clinics located around Bangkok with a license for possession and utilization of anorectic drugs, the psychotropic substances in category II. Body mass index was calculated at the time of survey. The prevalence of obesity, based on the proposed classification by body mass index in Asian adults, among the participants was 23.2% (95% CI: 18.0%-28.4%). Only 79 (31.6%) and 72 (28.8%) reported having low-calorie diets and increasing physical activities, respectively, both of which were recommended as the main part of comprehensive weight control. Cosmetic purpose was the main reason given by most participants (84.0%) to enter current weight-control treatments. Interestingly, most of the non-obese individuals (82.8%) misperceived themselves as being obese. These findings suggested that the inappropriate use of anorectic drugs among Thai women was a significant public health concern. Misperception of bodyweight status may contribute to the misuse of such anorectic drugs. The result would alarm the Food and Drug Administration, Ministry of Public Health to evaluate and revise the measures of the anorectic drug disposal. Further qualitative methods are recommended to investigate for body-image misconception, weight-control behavior among various populations. PMID- 15822545 TI - Effectiveness of constraint-induced movement therapy in chronic stroke patients. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness of constraint-induced movement therapy (CIMT) in dexterity with Action Research Arm Test (ARA test), hand grip strength, pinch strength of affected upper extremity in chronic stroke patients. MATERIAL AND METHOD: An observer-blinded randomized control trial, 69 chronic stroke patients were allocated either to constraint induced movement technique (n = 33) or conservative treatment (n = 36). The CIMT group received 6 hours of daily affected-upper-extremity training and restrained unaffected upper extremities for 5 days per week, totally 2 weeks. The control group received bimanual-upper-extremity training by conservative neurodevelopmental technique without restrained unaffected upper extremities for 2 weeks. RESULTS: The CIMT group had ARA scores, pinch strength of affected upper extremities statistically significant higher than the control group at p < 0.05, but the hand grip strength had no statistically significant difference, p > 0.05. CONCLUSIONS: CIMT of unaffected upper extremities has an advantage for chronic stroke patients which may be an efficacious technique of improving motor activity and exhibiting learned nonuse. PMID- 15822546 TI - Anesthesia for endoscopic retrograde cholangiopancreatography (ERCP) from 1999- 2003 in Siriraj Hospital: a retrospective study. AB - BACKGROUND: Endoscopic retrograde cholangio-pancreatography (ERCP) is another treatment option for hepato-biliary tract abnormalities. The authors studied anesthetic data as a basis for further research. METHOD: Retrospectively analyzed the patients on whom ERCP had been performed during the period of January, 1999 to November, 2003 in Siriraj Hospital. The patients' characteristics, preanesthetic problems, anesthetic techniques, anesthetic agents, anesthetic time, ERCP procedure and complications were assessed. RESULTS: There were 2,144 patients who received the procedure during study period The age group of 50-69 years was the highest one (46.9%). Most patients had ASA class 11 (54.7%). The diagnosis were stone (40.3%), tumor (34.0%), hepato-biliary tract infection (8.1%) and others (17.6%). Hypertension, diabetes mellitus and hematologic diseases were the most common preanesthetic problems. Total intravenous anesthesia (TIVA) was the anesthetic technique mainly employed (96.4%). Anesthetic agents were mainly administered with propofol, midazolam and fentanyl. The mean anesthetic time was 40.0+/-18.5 minutes. The indications for ERCP procedures were diagnostic (18.9%), stone removal (37.1%), stent removal and/or insertion (35.3%) and others (8.7%). The most frequent anesthetic complication was hypotension. CONCLUSION: During anesthetic management for ERCP, special techniques or drugs in anesthesia are not routinely required, however, the anesthetic personnel had to optimize the patient's condition for safety and there should be an awareness of complications. PMID- 15822547 TI - Diagnostic criteria for atopic dermatitis in Thai children. AB - Atopic dermatitis (AD) is a common skin disease in Thai children. There is no clinical or laboratory gold standard for the diagnosis. It is generally based on the guideline proposed by Hanifin and Rajka. Many studies have shown that some criteria are probably not all that significant in making the diagnosis. This study was designed to evaluate the frequency and diagnostic significance of clinical features of AD in Thai children. The authors studied 108 patients with AD and 103 controls including patients with other skin diseases. The AD group consisted of 60 girls and 48 boys. The mean age was 60.3+/-36.1 months. All previously proposed features were evaluated and the difference infrequency was tested with the chi-square test. History of pruritus, rash on typical distribution, chronically relapsing course, duration more than 6 months, personal or family history of atopy, age of onset before 2 years, recurrent conjunctivitis, itch when sweating, intolerance to rough textile, food and milk intolerance, history of dry skin, seasonal variation, visible dermatitis, dermatitis of a typical distribution, xerosis, ichthyosis vulgaris, foot dermatitis, Dennie-Morgan infraorbital fold, orbital darkening, periorbital dermatitis, pityriasis alba, peri-auricular dermatitis, anterior neck fold, truncal dermatitis, perifollicular accentuation, white dermographism and diffuse scaling of scalp were all significantly more frequent in AD (p < 0.05). A minimum set of diagnostic criteria for AD was derived by using multiple stepwise logistic regression technique. It consisted of history of itchy rash, history of flexural dermatitis, chronicity more than 6 months, and visible xerosis, periorbital dermatitis and perifollicular accentuation. PMID- 15822548 TI - Lactose intolerance in Thai adults. AB - Lactose intolerance is common in Thai adults who ingest cow's milk but its incidence has not been clearly defined The authors evaluated 45 volunteers (15 males, 35 females), aged 21-31 yrs old, who drank one 240-ml box of milk daily. A Lactose tolerance test was performed using a breath-hydrogen test (BHT) after oral intake of 25 g of lactose dissolved in 250 ml of water The presence of gastrointestinal symptoms of lactose intolerance, flatulence, abdominal pain and diarrhea, were recorded Twenty-one subjects (47%) were categorized as lactose malabsorbers and intolerant, two subjects (4%) were malabsorbers but tolerant, and 22 of 45 (49%) were absorbers and tolerant. The incidence of lactose malabsorption was, thus, 51%; symptoms of intolerance were found in 21 of the 23 malabsorbers, making the incidence of lactose intolerance 47%. In the lactose malabsorbant and intolerant group, the more breath-hydrogen (H) the more symptoms observed All subjects who had a negative breath-H2 test had no symptoms. The breath-H2 test should be used as a standard method to evaluate lactose absorption and lactose tolerance. The incidence of lactose intolerance has decreased from the past and the symptoms are not so severe that the people limit the consumption of milk since it is a major source of food containing good quality of protein and calcium. PMID- 15822549 TI - Changes in erythrocyte contents of potassium, sodium and magnesium and Na, K-pump activity after the administration of potassium and magnesium salts. AB - Low potassium and magnesium status and decreased Na, K-pump activity is an endemic condition among rural Northeast Thais. The authors examined the effect of supplementing potassium and magnesium on erythrocyte potassium, sodium and magnesium content and on Na, K-pump activity. Rural Northeast Thai renal stone patients (62) were recruited, divided into four groups and supplemented for one month with potassium chloride (Group1, n = 16), potassium-sodium citrate (Group2, n = 15), chelated magnesium (Group 3, n =16) and potassium-magnesium citrate (Group 4, n =15) in order to achieve 40 mmol potassium, 10 mmol magnesium and 60 mmol citrate daily. After supplementation with potassium (Groups 1, 2 and 4), plasma potassium and Na, K-pump activity rose significantly in Groups 1, 2 and 4, but erythrocyte potassium rose only in Groups 2 and 4. When supplementing elemental magnesium (Groups 3 and 4), the chelated magnesium caused a significant increase in plasma potassium, erythrocyte potassium, sodium and magnesium without a significant increase in Na, K-pump activity. By contrast, potassium-magnesium citrate caused a significant increase in erythrocyte potassium and magnesium and Na, K-pump activity, but depressed erythrocyte sodium. These results suggest the forms of potassium and /or magnesium salts being supplemented should be considered because they affect erythrocyte potassium, sodium and magnesium content and Na, K-pump activity differently. PMID- 15822550 TI - Randomized controlled trial between surgery and aspiration combined with methylprednisolone acetate injection plus wrist immobilization in the treatment of dorsal carpal ganglion. AB - Dorsal carpal ganglion is one of the most common benign tumors of the hand. Many treatment modalities had been proposed and yielded the same success rate. In the present study, the authors conducted a randomized controlled trial to compare the success of treatment between surgery and aspiration combined with methylprednisolone acetate injection plus wrist immobilization. There were 28 patients enrolled and randomized into two treatment groups. Only 24 patients were available for the final follow up at 6 months. According to the present study, dorsal carpal ganglion was more common in females (83.3%) at an average age of 30 years. Pain was the most common presenting symptom (58.3%). The success by excision was 81.8% and by aspiration combined with methylprednisolone acetate injection plus wrist immobilization was 38.46%. The p-value was 0.047 by Fisher exact test. The present study has clearly shown that surgical excision gave a better success rate in the treatment of dorsal carpal ganglion. PMID- 15822551 TI - Oculopharyngodistal myopathy in a Thai family. AB - There has been controversy whether oculopharyngodistal myopathy (OPDM) commonly seen in Japan is a distinct disease entity or a variant of oculopharyngeal muscular dystrophy (OPMD) initially described in French-Canadians and has since been reported in other ethnic groups. Both diseases have autosomal dominant inheritance and OPDM patients are clinically similar to OPMD with slowly progressive ptosis, ophthalmoplegia and dysphagia except that most of the former usually have distal as opposed to proximal weakness and most of them are genetically different from the latter The authors report here 2 siblings with clinical features of OPDM. This entity is rare outside Japan and this is the first family to be reported from Thailand PMID- 15822552 TI - Ruptured mature cystic teratomas mimicking advanced stage ovarian cancer: a report of 2 cases study. AB - Spontaneous rupture of mature cystic teratoma is uncommon. Chronic rupture of teratomas may result in granulomatous peritonitis. In rare cases, the clinical presentation and the intraoperative findings may mimic those of advanced stage ovarian cancers. Two cases of ruptured mature cystic teratomas mimicking malignancy are reported in 50- and 53-year-old patients. Both presented with abdominal distension and weight loss. Elevation of serum CA-125 (233 unit/ml) was detected in one case. Intraoperatively, nodular thickening of the omentum with marked peritoneal adhesion was observed in both cases. Ascites of 1500 ml was present in one case. Pathologic examination in both cases showed ovarian mature cystic teratomas with peritoneal granulomatous inflammations response to the released tumor content. Both patients were followed for 44 and 12 months and were free of symptoms without additional treatment other than surgery. Intraoperative pathologic consultation can help confirm the benign diagnosis and unnecessary major operation for malignancy can be avoided. PMID- 15822553 TI - Environmental reduplication in a patient with right middle cerebral artery occlusion. AB - Environmental reduplication or reduplicative paramnesia is one of the content specific delusions (CSD) which is characterized by reduplication of places. CSD has been reported in focal and diffuse cerebral disorders. A focal lesion such as frontal lobes and the right hemispheric lesion have been documented The authors describe a 66 year-old woman who had a delusion of misidentification for place one month after right middle cerebral artery occlusion. The patient did not have any history of schizophrenia or other psychiatric diseases. The patient believed that her car, furniture and house were duplicated. She also mentioned that her son and friends tried to takeover all of her properties and told everyone that she was insane. The prominent cortical signs were tactile and visual neglect. Neuropsychological assessments revealed poor attention but she had neither confusion nor dementia. Clock drawing and construction tests revealed visuospatial impairment which was compatible with non-dominant hemispheric abnormality. MRI showed evidence of cerebral infarction in the right middle cerebral artery territory. Only one similar patient who had an intracerbral hematoma of the right frontal lobe has been reported in the literature. The role of occipito-parietal and fronto-temporal lobes or their connections in environmental reduplication is proposed. PMID- 15822554 TI - Paradigm shift in rabies control: a system approach. AB - Government centered approach in rabies prevention and control has not led to rabies safe community, both in urban and rural areas. Government services and organizations should recognize that only joint collaboration among households and community can lead to a successful rabies control mission. Dog owners should have been more actively involved in the prevention and solving of rabies problem. Promotion and support of government organizations should aim to strengthen the communities to become self reliance in diseases control and health care services. Only by strengthening community roles result in a more sustainable community disease control program. PMID- 15822555 TI - Questionable incidence of Dalmatian urolithiasis. PMID- 15822556 TI - Diagnosing equine pars intermedia dysfunction: are we there yet? PMID- 15822557 TI - Recognition and classification of dysmyelopoiesis in the dog: a review. AB - Dysmyelopoiesis is defined as a hematologic disorder characterized by the presence of cytopenias in the blood and dysplastic cells in one or more hematologic cell lines in the blood or bone marrow. The causes of dysmyelopoiesis include acquired mutations in hematopoietic stem cells (i.e., myelodysplastic syndromes [MDSs]), congenital defects in hematopoiesis, and dysmyelopoietic conditions associated with various disease processes, drug treatments, or toxin exposure. Two major subtypes of MDSs (i.e., MDS with refractory cytopenias and MDS with excess myeloblasts) have been described that differ in clinical presentation, response to treatment, and survival time. The most frequently occurring causes of secondary dysmyelopoiesis include immune-mediated hematologic diseases, lymphoid malignancies, and exposure to chemotherapeutic drugs. Differentiation of the various causes of dysmyelopoiesis is essential for establishing an appropriate therapeutic plan and for determining prognosis. PMID- 15822558 TI - Early biochemical and clinical responses to cobalamin supplementation in cats with signs of gastrointestinal disease and severe hypocobalaminemia. AB - Domestic cats with small intestinal disease may develop cobalamin deficiency because of reduced small intestinal uptake of this vitamin. This study assessed the impact of cobalamin deficiency on biochemical and clinical findings in cats with intestinal disease. Nineteen pet cats, all with severe hypocobalaminemia (< or =100 ng/L) and histories of gastrointestinal signs, were studied. Cats received cobalamin, 250 microg SC once weekly, for 4 weeks. Biochemical indices of cobalamin availability (e.g., serum methylmalonic acid, homocysteine, and cysteine concentrations), serum feline trypsinlike immunoreactivity (fTLI) and serum folate concentrations, and clinical findings were recorded at the start of the study and after 4 weeks of cobalamin therapy. Serum methylmalonic acid (MMA) concentrations (median; range) decreased after cobalamin supplementation (5373.0; 708.5-29,329.0 versus 423.5; 214.0-7219.0 nmol/L, P < .0001). Serum homocysteine concentrations were not significantly altered (mean +/- SD 8.2 +/- 2.9 versus 10.3 +/- 4.5 micromol/L, P = .1198), whereas cysteine concentrations increased significantly (122.3 +/- 38.8 versus 191.5 +/- 29.4 micromol/L, P < .0001). Mean body weight increased significantly after cobalamin therapy (3.8 +/- 1.1 versus 4.1 +/- 1 kg, P < .01), and the average body weight gain was 8.2%. Significant linear relationships were observed between alterations in serum MMA and fTLI concentrations and the percentage body weight change (P < .05 for both, Pearson r2 = 0.26 and 0.245, respectively). Mean serum folate concentration decreased significantly (mean +/- SD 19 +/- 5 microg/L versus 15.4 +/- 6.2 microg/L, P < .001). Reduced vomiting and diarrhea were observed in 7 of 9 and 5 of 13 cats, respectively. These results suggest that cobalamin supplementation in cats with small intestinal disease and severe hypocobalaminemia is associated with normalization of biochemical test results and improvements in clinical findings in most affected cats. PMID- 15822559 TI - Bronchodilators in bronchoscopy-induced airflow limitation in allergen-sensitized cats. AB - This study investigated the effect of bronchoscopy and bronchoalveolar lavage (BAL) on respiratory function, determined by barometric whole-body plethysmography (BWBP), of healthy and allergen-sensitized cats. Furthermore, the efficacy of inhaled bronchodilators in preventing changes in respiratory function was determined. For test 1, 18 healthy experimental cats were investigated on day 1 by BWBP. On day 2, the cats underwent BWBP after sedation (medetomidine), after anesthesia induction (propofol), and after bronchoscopy and BAL. Enhanced pause (Penh) was significantly increased after bronchoscopy and BAL (1.64 +/- 0.17 versus 1.23 +/- 0.07, P < .05). For test 2, 6 cats were sensitized to ovalbumin (OVA), 6 cats were sensitized to Ascaris suum (AS), and 6 cats served as controls. On day 0, OVA- and AS-sensitized cats underwent an inhaled allergen challenge, whereas controls were exposed to saline. On days 1 and 2, the same protocol as described for test 1 was repeated. Post-BAL Penh of the AS-sensitized cats was significantly higher than at test 1 (2.28 +/- 0.22 versus 1.69 +/- 0.33, P < .05) and was correlated with BAL fluid neutrophil count (r = 0.55, P < .05). During tests 3, 4, and 5, the same protocol as used for test 2 was applied to each cat group, with the animals being randomly treated before sedation with inhaled salbutamol (200 microg), ipratropium bromide (40 microg), or a combination of both (200 + 40 microg). Post-BAL Penh of the AS-sensitized group was significantly decreased after the salbutamol + ipratropium bromide treatment (1.56 +/- 0.18 versus 2.28 +/- 0.22, P < .05). This study suggests that bronchoscopy and BAL induce airflow limitation in cats, which is more severe in the presence of lower airway inflammation. Inhaled salbutamol + ipratropium bromide reduce BAL-induced bronchoconstriction in AS-challenged cats and might be recommended as preventive treatment of asthmatic cats undergoing bronchoscopy. PMID- 15822560 TI - Somatostatin receptor imaging in vivo by planar scintigraphy facilitates the diagnosis of canine insulinomas. AB - Somatostatin receptors expressed by insulinomas in 5 dogs were imaged in vivo by means of indium in 111 pentetreotide (OctreoScan) scintigraphy. The diagnosis in each dog was supported by the presence of hypoglycemia (<60 mg/dL), hyperinsulinemia (>20 microU/mL), and histopathologic review of neoplastic tissue. All insulinomas expressed high-affinity somatostatin receptors of subtype sst2, as shown by receptor autoradiography in vitro using 125I-[tyrosine3] octreotide and 125I-[leucine8, Dtryptophan22, tyrosine25]-somatostatin-28 with an sst2 subtype-selective analogue. Scintigrams were obtained at 1, 4, 12, and 24 hours after the i.v. administration of 74-222 MBq of OctreoScan to each patient. Abnormal foci of activity were 1st observed from 1 hour after administration of the radioligand in dog 3, to 24 hours after its administration in dog 4; in dogs 1 and 2, abnormal foci of activity were visible from 12 hours. Dog 5 showed a questionable abnormal focus of activity at 12 hours, but not at 24 hours. Scintigraphy enabled accurate prediction of the anatomical location of the primary tumor in 1 of 4 dogs, but was unable to differentiate a right- from a left-pancreatic lobe tumor, or vice versa, in 3 dogs; the 5th dog had equivocal results. 111In-pentetreotide scintigraphy is a useful diagnostic adjunct to the clinical evaluation of the insulinoma patient, but is unable to localize the tumor in some cases. PMID- 15822561 TI - Tylosin-responsive chronic diarrhea in dogs. AB - Fourteen dogs had shown chronic or intermittent diarrhea for more than 1 year. Diarrhea had been successfully treated with tylosin for at least 6 months but recurred when treatment was withdrawn on at least 2 occasions. Tylosin-responsive diarrhea (TRD) affects typically middle-aged, large-breed dogs and clinical signs indicate that TRD affects both the small and large intestine. Treatment with tylosin eliminated diarrhea in all dogs within 3 days and in most dogs within 24 hours. Tylosin administration controlled diarrhea in all dogs, but after it was discontinued, diarrhea reappeared in 12 (85.7%) of 14 dogs within 30 days. Prednisone given for 3 days did not completely resolve diarrhea. Probiotic Lactobacillus rhamnosus GG did not prevent the relapse of diarrhea in any of 9 dogs so treated. The etiology of TRD, a likely form of antibiotic-responsive diarrhea (ARD) is unclear. The following reasons for chronic diarrhea were excluded or found to be unlikely: parasites, exocrine pancreatic insufficiency, inflammatory bowel disease, small intestinal bacterial overgrowth, enteropathogenic bacteria (Salmonella spp., Campylobacter spp., Yersinia spp., or Lawsoni intracellularis), and Clostridium perfringens enterotoxin and Clostridium difficile A toxin. A possible etiologic factor is a specific enteropathogenic organism that is a common resident in the canine gastrointestinal tract and is sensitive to tylosin but difficult to eradicate. Additional studies are required to identify the specific cause of TRD. PMID- 15822562 TI - Hereditary spectrin deficiency in Golden Retriever dogs. AB - Spectrin deficiency with increased erythrocyte osmotic fragility (OF) is a hallmark of hereditary spherocytosis, which is the most common congenital hemolytic anemia in humans of northern European ancestry. A radioimmunoassay revealed that erythrocyte spectrin concentration was 50-65% of normal in 5 adult Golden Retriever dogs, which had recovered from hemolytic anemia but whose OF had persistently remained increased. OF also was increased and spectrin concentration was decreased (60-73%) in 10 dogs of an apparently healthy family of 19 Golden Retrievers related to a proband. Pedigree analysis revealed autosomal dominant inheritance. In addition, OF was increased in 23 (17%) of 134 randomly chosen Golden Retrievers with nonhematologic diseases. In these Golden Retrievers, the spectrin concentration was decreased in 5 dogs with increased OF and within the reference range in 6 dogs with normal OF, indicating that in this population spectrin deficiency and increased OF are highly associated (P < .002). Considering these patients a representative sample of the Golden Retriever population in the Netherlands, spectrin deficiency may occur in 11.2-24.6% of Dutch Golden Retrievers (confidence level = 0.95). In blood smears, spherocytes were recognized only in dogs with immune-mediated anemia. At scanning electron microscopy, blood from spectrin-deficient Golden Retrievers showed slight crenation when fixed freshly but abundant echinospherocytes after 24 hours of incubation. We conclude that occult autosomal dominant spectrin deficiency occurs in dogs and is frequent in Dutch Golden Retrievers. It is not clear whether spectrin deficiency in Golden Retrievers may result in hemolytic anemia, as in humans. PMID- 15822563 TI - The immunophenotype of peripheral blood lymphocytes in clinically healthy dogs and dogs with lymphoma in remission. AB - Lymphoma is a common cancer of dogs that frequently is treated with chemotherapy or radiation therapy. Response to therapy is variable and currently available diagnostic tests do not reliably predict response to therapy. Treatment for lymphoma often results in lymphopenia, but it is unknown whether the changes in circulating lymphocytes result from generalized or specific reduction of lymphocytes. In this study, blood lymphocytes from 12 clinically healthy dogs, 10 dogs in remission because of treatment for B-cell lymphoma, and 8 dogs in remission from T-cell lymphoma were analyzed by flow cytometry by using a panel of 20 antibodies reactive with canine leukocyte antigens. Results identified similar lymphocyte parameters in treated dogs regardless of the type of lymphoma. Treated dogs had >50% reduction in blood lymphocyte concentration, and an absolute decrease in most subsets of lymphocytes. Both groups of treated dogs had relative increases in the proportion of CD3+, T-cell receptor (TCR)alphabeta+, and CD90+ lymphocytes, and a decreased proportion of CD45RA+ cells. In addition, dogs with T-cell lymphoma in remission had a significant increase in the proportion of CD49d+ lymphocytes. These findings were interpreted as representing likely suppression of lymphocyte regeneration by chemotherapy, with a relative increase in the proportion of memory over naive lymphocytes. Lack of correlation with the T- or B-cell origin of the initial lymphoma suggested that, by using flow cytometric methods, residual circulating neoplastic cells could not be detected. However, the changes in the lymphocyte profile of dogs treated with chemotherapy may have relevance to their immunocompetence. PMID- 15822564 TI - Characterization of the mutations causing hemophilia B in 2 domestic cats. AB - The purpose of the present study was to determine the normal sequence for the gene encoding factor IX in cats and to characterize the genetic basis for hemophilia B in 2 unrelated male, domestic, mixed-breed cats. Genomic DNA sequence for the entire coding region of the factor IX gene was determined in the affected cats and compared to the sequence obtained from a healthy cat. The factor IX gene in cats encodes a mature protein consisting of 420 amino acids, unlike genes in humans and dogs that encode 415 and 413 amino acid proteins, respectively. Affected cat 1 had a single nucleotide change in exon 8 at the 1st nucleotide position of the codon encoding an arginine (CGA to TGA) at amino acid position 338. This mutation would be predicted to result in the appearance of a premature stop codon in the portion of the gene encoding much of the catalytic domain of the protein. Affected cat 2 had a single nucleotide change in exon 4 at the 2nd nucleotide position of the codon encoding amino acid 82 (TGT to TAT), which would be predicted to result in the substitution of a tyrosine for a cysteine. This substitution would likely result in disruption of a disulfide bond crucial to normal protein structure and function. This study represents the 1st time hemophilia B has been characterized at the molecular level in cats. PMID- 15822565 TI - Cosegregation of a factor VIII microsatellite marker with mild hemophilia A in Golden Retriever dogs. AB - Mild hemophilia A (factor VIII deficiency) was diagnosed in Golden Retrievers and pedigree studies were undertaken to test the cosegregation of an intragenic factor VIII marker with the disease phenotype. The study population consisted of 30 client-owned dogs (22 males and 8 females). Hemophilic males (n = 12) typically demonstrated prolonged bleeding after trauma or surgery rather than spontaneous hemorrhagic events. The affected males had a proportionate reduction in factor VIII coagulant activity (mean FVIII:C = 4%) and factor VIII protein concentration (mean FVIII:Ag = 3%). Twenty-five dogs (10 affected males, 8 clear males, 2 obligate carrier dams, and 5 suspect carrier daughters) were genotyped for a factor VIII microsatellite marker, with allele size assigned by an automated capillary electrophoresis system. Five distinct marker alleles were present in the study pedigree and a 300-base pair allele was found to segregate with the hemophilia A phenotype. The inheritance of the hemophilia-associated allele defined carrier status for 5 suspect daughters of obligate carrier dams. The limitations inherent to linkage analyses (i.e., lack of access to key family members and homozygosity at the marker locus) did not preclude carrier detection in this pedigree. We conclude that genotype analysis for the intragenic factor VIII marker can aid in control of canine hemophilia A through enhanced carrier detection. PMID- 15822566 TI - Ulcerative dermatitis, thrombocytopenia, and neutropenia in neonatal foals. AB - This report describes transient ulcerative dermatitis, severe thrombocytopenia, and mild neutropenia in 6 foals from 4 mares from geographically diverse regions of the United States. The foals presented at <4 days of age with oral and lingual ulcers, and crusting and erythema around the eyes, muzzle, and perineal, inguinal, axillary, trunk, and neck regions. There was a severe thrombocytopenia (0-30,000 platelets/microL), leukopenia (1900-3200 white blood cells/microL), and mild neutropenia (500-1800 neutrophils/microL). Four of the 6 foals had petechiae and ecchymotic hemorrhages and 3 had bleeding tendencies. Results of examination of a bone marrow biopsy from 1 foal were normal and results of a platelet surface immunoglobulin test in another were negative. Histopathology of the skin in all foals showed subepidermal clefting with subjacent vascular dilation, dermal hemorrhage, and superficial papillary necrosis. The foals were treated supportively with broad-spectrum antibiotics (5/6), corticosteroids (3/6), gastric ulcer prophylaxis (6/6), whole-blood transfusion (4/6), and platelet-rich plasma (1/6). The skin lesions and thrombocytopenia (>50,000 platelets/microL) improved in 2 weeks (4/6). Two foals had a decline in their platelet counts when the steroids were decreased and needed protracted treatment. All foals survived and were healthy as yearlings. Two mares that had 2 affected foals each, upon subsequent pregnancies to different stallions, had healthy foals when an alternate source of colostrum was given. The findings in the cases in this report suggest a possible relationship between colostral antibodies or some other factor in the colostrum and the thrombocytopenia and skin lesions, although further investigation is warranted to confirm or refute this hypothesis. PMID- 15822567 TI - Variation in plasma adrenocorticotropic hormone concentration and dexamethasone suppression test results with season, age, and sex in healthy ponies and horses. AB - The purpose of this study was to evaluate the variation in plasma adrenocorticotropic hormone (ACTH) concentration and dexamethasone suppression test (DST) results with season, age, and sex in healthy, pony mares (n=15) and pony stallions (n=14) living under semiferal conditions and horse mares (n=10) living at pasture. Plasma ACTH concentrations were measured in September 2002, and in January, May, and September 2003. DSTs were performed in January and September 2003. Plasma ACTH concentrations in September 2002 and September 2003 were similar and were significantly greater than in January and May (P < .001). Plasma ACTH concentration was within the reference range for 38 (97%) of 39 subjects in January, for 39 (100%) of 39 subjects in May, for 2 (5%) of 39 subjects in September 2002, and for 3 (8%) of 39 subjects in September 2003. DST results were within the reference range in all subjects in January and were within the reference range for 29 (74%) of 39 subjects in September 2003. Plasma cortisol concentration at the end of the DST was significantly greater in September than in January (P = .002). Age was positively correlated with plasma ACTH and plasma cortisol concentration at the beginning and end of the DST Within the same season, plasma ACTH concentration in pony mares, pony stallions, and horse mares was not significantly different (P > .05). Seasonal changes in plasma ACTH concentration and DST results should be considered when interpreting endocrine test results. PMID- 15822568 TI - Alterations in serum parathyroid hormone and electrolyte concentrations and urinary excretion of electrolytes in horses with induced endotoxemia. AB - Hypocalcemia and hypomagnesemia are common in horses with sepsis and endotoxemia. We hypothesize that endotoxemia triggers a systemic inflammatory response that results in hypocalcemia and hypomagnesemia. The goal of this study was to determine the effect of endotoxin (lipopolysaccharide [LPS]) administration to healthy horses on serum parathyroid hormone (PTH), ionized calcium (Ca2+) and total calcium (tCa), ionized magnesium (Mg2+) and total magnesium (tMg), phosphate (Pi), potassium (K+), sodium (Na+), chloride (Cl-), and insulin concentrations, and on the urinary excretion of these electrolytes. Twelve mares were infused with Escherichia coli LPS (30 ng/kg/h i.v.) for 1 hour. Six mares were infused with saline (controls). In LPS-infused horses, heart rate increased significantly from (mean +/- SD) 40.0 +/- 1.3 to 70.0 +/- 9.0 beats/min, respiratory rate from 12.7 +/- 1.0 to 21.1 +/- 3.0 breaths/min, body temperature from 37.4 +/- 0.3 to 38.9 +/- 0.6 degrees C, and tumor necrosis factor-alpha concentrations from 6.6 +/- 3.5 to 507 +/- 260 pg/mL (P < .05). White blood cell count decreased significantly from 7570 +/- 600 to 1960 +/- 560 cells/ microL. Serum concentrations of Ca2+ decreased from 6.5 +/- 0.3 to 6.0 +/- 0.3 mg/dL, of Mg2+ from 0.53 +/- 0.06 to 0.43 +/- 0.04 mM, of tMg from 0.78 +/- 0.05 to 0.62 +/ 0.08 mM, of K+ from 4.3 +/- 0.4 to 3.0 +/- 0.5 mEq/L, and of Pi from 3.4 +/- 0.5 to 1.7 +/- 0.5 mg/dL (all P < .05). PTH increased significantly from 1.3 +/- 0.4 to 6.0 +/- 5.2 pM; however, in some horses (n=2), PTH did not increase despite hypocalcemia. Insulin increased significantly from 9.4 +/- 3.6 to 50.5 +/- 9.6 microIU/mL (n=3). Urinary fractional excretion of Ca2+ decreased significantly from 4.7 +/- 1.4 to 1.7 +/- 1.2%, of Mg2+ from 36.6 +/- 6.5 to 11.7 +/- 7.3%, and of K+ from 37.9 +/- 11.3 to 17.7 +/- 6.2%. Fractional excretion of Pi increased from 0.02 +/- 0.02 to 0.14 +/- 0.07% and of Na+ from 0.26 +/- 0.13% to 1.2 +/- 0.5%. No changes were found in serum tCa, Na+, and Cl- concentrations. In conclusion, endotoxemia in horses resulted in electrolyte abnormalities that included hypocalcemia, hypomagnesemia, hypokalemia, hypophosphatemia, and increased serum PTH and insulin concentrations. PMID- 15822569 TI - Acute clinical, hematologic, serologic, and polymerase chain reaction findings in horses experimentally infected with a European strain of Anaplasma phagocytophilum. AB - Six horses were experimentally infected by administration of horse blood containing a Swedish strain of Anaplasma phagocytophilum. The polymerase chain reaction (PCR) signal was consistently detected 2-3 days before appearance of clinical signs and persisted 4-9 days beyond abatement of clinical signs, whereas diagnostic inclusion bodies were 1st noted on average 2.6 +/- 1.5 (SD) days after onset of fever. Clinical signs and hematologic changes were largely indistinguishable from those previously reported for diseases caused by A phagocytophilum (formerly Ehrlichia equi--"Californian agent") and the human derived human granulocytic ehrlichiosis agent. Horses 1st demonstrated antibody response 12-16 days after inoculation, 2 cases of which were still febrile, and serotiters rapidly peaked within 3-7 days of clinical illness. One horse died during the acute stage of disease, but initial clinical signs and hematologic changes were similar to those of other infected horses. This report shows that, despite minor genetic differences, a European equine-derived strain of A. phagocytophilum may be similar in pathogenicity to the Californian agent. The PCR used holds promise to widen the diagnostic window and would also be diagnostic during the initial days of clinical disease when inclusions in neutrophils in blood smears are not yet apparent. PMID- 15822570 TI - Effects of intravenous hyperosmotic sodium bicarbonate on arterial and cerebrospinal fluid acid-base status and cardiovascular function in calves with experimentally induced respiratory and strong ion acidosis. AB - The objectives of this study were to determine the effects of hyperosmotic sodium bicarbonate (HSB) administration on arterial and cerebrospinal fluid (CSF) acid base balance and cardiovascular function in calves with experimentally induced respiratory and strong ion (metabolic) acidosis. Ten healthy male Holstein calves (30-47 kg body weight) were instrumented under halothane anesthesia to permit cardiovascular monitoring and collection of blood samples and CSE Respiratory acidosis was induced by allowing the calves to spontaneously ventilate, and strong ion acidosis was subsequently induced by i.v. administration of L-lactic acid. Calves were then randomly assigned to receive either HSB (8.4% NaHCO3; 5 ml/kg over 5 minutes, i.v.; n=5) or no treatment (controls, n=5) and monitored for 1 hour. Mixed respiratory and strong ion acidosis was accompanied by increased heart rate, cardiac index, mean arterial pressure, cardiac contractility (maximal rate of change of left ventricular pressure), and mean pulmonary artery pressure. Rapid administration of HSB immediately corrected the strong ion acidosis, transiently increased arterial partial pressure of carbon dioxide (P(CO2)), and expanded the plasma volume. The transient increase in arterial P(CO2) did not alter CSF P(CO2) or induce paradoxical CSF acidosis. Compared to untreated control calves, HSB-treated calves had higher cardiac index and contractility and a faster rate of left ventricular relaxation for 1 hour after treatment, indicating that HSB administration improved myocardial systolic function. We conclude that rapid i.v. administration of HSB provided an effective and safe method for treating strong ion acidosis in normovolemic halothane anesthetized calves with experimentally induced respiratory and strong ion acidosis. Fear of inducing paradoxical CSF acidosis is not a valid reason for withholding HSB administration in calves with mixed respiratory and strong ion acidosis. PMID- 15822571 TI - Obstructive renal cyst in a dog: ultrasonography-guided treatment using puncture aspiration and injection with 95% ethanol. PMID- 15822572 TI - Spontaneous hypoadrenocorticism in a dog after a diagnosis of hyperadrenocorticism. PMID- 15822573 TI - Intermittent claudication in an Afghan hound due to aortic arteriosclerosis. PMID- 15822574 TI - Double-outlet right ventricle in an Angus calf. PMID- 15822575 TI - Mitral valve endocarditis after dental prophylaxis in a dog. PMID- 15822576 TI - Radiation therapy for the treatment of urethral sarcoma in a cria. PMID- 15822577 TI - Determination of abomasal emptying rate in suckling calves by use of nuclear scintigraphy and acetaminophen absorption. AB - OBJECTIVE: To develop nuclear scintigraphic and acetaminophen absorption methods for measuring abomasal emptying rate in suckling calves. ANIMALS: 9 male Holstein Friesian calves < 30 days old. PROCEDURE: Calves were fed 2 L of milk replacer, fresh cow's milk, or an isoosmotic (150mM) solution of NaCl, NaHCO3, or sodium acetate containing technetium TC 99m-diethylenetriamine-pentaacetic acid (370 MBq) and acetaminophen (50 mg/kg). Right lateral scintigraphic images and venous blood samples were obtained periodically after feeding. Two power exponential equations were fitted to the scintigraphic data, and 3 pharmacokinetic models were fitted to the acetaminophen concentration-time data. RESULTS: Data from 32 feedings were analyzed, with half emptying time for scintigraphic evaluation ranging from 29 to 202 minutes. Siegel's modified power exponential equation provided a better description of the scintigraphic data than did Elashoff's power exponential equation. The first derivative of Siegel's modified power exponential formula provided the best pharmacokinetic model for the acetaminophen absorption data. Time to maximal acetaminophen concentration (Tmax) provided the most accurate index of abomasal emptying rate from the acetaminophen concentration time data. CONCLUSIONS AND CLINICAL RELEVANCE: Abomasal emptying rate is best determined in healthy suckling calves by use of Siegel's modified power exponential equation to model the scintigraphic data. Pharmacokinetic calculation of Tmax from the acetaminophen absorption curve provides an alternative method for determining abomasal emptying rate in healthy suckling calves that is accurate, inexpensive, practical, and safe. However, it is unclear whether diarrhea would alter the acetaminophen absorption curve in calves. PMID- 15822578 TI - Assessment of the reliability of plasma electrophoresis in birds. AB - OBJECTIVE: To determine the reliability of plasma electrophoresis (EPH) in psittacine birds. ANIMALS: 93 psittacine birds. PROCEDURE: Jugular venipuncture was performed on 93 awake psittacine birds. The plasma was centrifuged, separated, aliquoted into duplicate samples, frozen, and sent to 2 commercial laboratories that routinely perform avian EPH. Samples from 51 birds were sent to laboratory A, and samples from 42 birds were sent to laboratory B. The reliability of EPH results within each laboratory was assessed, but not between laboratories. To determine the reliability (agreement between duplicate samples) of total protein, albumin, prealbumin, alpha1-, alpha2-, beta-, and gamma globulin concentrations, the intraclass correlation coefficient (r(i)) was calculated. RESULTS: Both laboratories had excellent agreement between samples for measurement of total protein concentration and only good agreement for albumin concentration. Except for the prealbumin concentration measured at laboratory B, both laboratories had poor agreement for all other values of the EPH. CONCLUSIONS AND CLINICAL RELEVANCE: These data indicate that plasma EPH for measuring prealbumin, alpha1-, alpha2-, beta-, and gamma-globulin concentrations may not be a reliable tool for assessing avian health. Small amounts of these proteins in birds plus human variation in reading the EPH curves may lead to variable results. Avian veterinarians should cautiously interpret results from plasma EPH assays for these protein fractions. PMID- 15822579 TI - Expression of equine glucose transporter type 4 in skeletal muscle after glycogen depleting exercise. AB - OBJECTIVES: To clone and sequence cDNA for equine insulin-responsive glucose transporter (glucose transporter type 4 [GLUT-4]) and determine effects of glycogen-depleting exercise and meal type after exercise on GLUT-4 gene expression in skeletal muscle of horses. SAMPLE POPULATION: Muscle biopsy specimens from 7 healthy adult horses. PROCEDURES: Total RNA was extracted from specimens, and GLUT-4 cDNA was synthesized and sequenced. Horses were exercised on 3 consecutive days. On the third day of exercise, for 8 hours after exercise, horses were either not fed, fed half of daily energy requirements as hay, or fed an isocaloric amount of corn. The GLUT-4 mRNA was determined by use of real-time reverse transcriptase-polymerase chain reaction in muscle biopsy specimens obtained before 3 consecutive days of exercise and within 10 minutes and 4, 8, and 24 hours after the third exercise bout. RESULTS: A 1,629-bp segment was sequenced, of which 1,530 bp corresponded to the coding region end encoded a protein of 509 amino acids. Expression of GLUT-4 gene increased by 2.3, 4.3, 3.3, and 2.6 times 10 minutes and 4, 8, and 24 hours after exercise, respectively, compared with that prior to exercise. No differences were observed in GLUT-4 gene expression among conditions of feed withholding, corn feeding, and hay feeding during the 8 hours postexercise. CONCLUSIONS AND CLINICAL RELEVANCE: Lack of increase of GLUT-4 gene expression after grain feeding and exercise may explain the apparently slower rate of glycogen synthesis after exercise in horses relative to that of other species. PMID- 15822580 TI - Effects of preanesthetic administration of morphine on gastroesophageal reflux and regurgitation during anesthesia in dogs. AB - OBJECTIVE: To determine the effect of morphine administered prior to anesthesia on the incidence of gastroesophageal reflux (GER) in dogs during the subsequent anesthetic episode. ANIMALS: 90 dogs (30 dogs/group). PROCEDURE: The randomized prospective clinical study included healthy dogs with no history of vomiting. Dogs were scheduled to undergo elective orthopedic surgery. Food was withheld for (mean+/-SD) 17.8+/-4.1 hours prior to induction of anesthesia. The anesthetic protocol included acepromazine maleate, thiopental, and isoflurane. Dogs were randomly selected to receive morphine at various dosages (0, 0.22, or 1.10 mg/kg, IM) concurrent with acepromazine administration prior to induction of anesthesia. A sensor-tipped catheter was used to measure esophageal pH, and GER was defined as a decrease in pH to < 4 or an increase to > 7.5. RESULTS: 40 dogs had acidic reflux, and 1 had biliary reflux. Proportions of dogs with GER were 8 of 30 (27%), 15 of 30 (50%), and 18 of 30 (60%) for morphine dosages of 0, 0.22, and 1.10 mg/kg, respectively. Mean duration of GER was 91.4+/-56.8 minutes. There was no significant association between GER and age, weight, vomiting after preanesthetic medication, administration of antimicrobials, or start of surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Most healthy dogs vomit after a large dose of morphine, but vomiting does not increase the likelihood of GER during the subsequent anesthetic episode. Administration of morphine prior to anesthesia substantially increases the incidence of GER during the subsequent anesthetic episode. PMID- 15822581 TI - Evaluation of the passive function of the biceps brachii muscle-tendon unit in limitation of shoulder and elbow joint ranges of motion in horses. AB - OBJECTIVE: To quantify the passive contribution of the biceps brachii muscle tendon unit to the limits of elbow joint extension during shoulder joint flexion in horses. SAMPLE POPULATION: Normal right forelimb specimens from 6 Thoroughbred cadavers. PROCEDURE: Specimens included the scapula, humerus, radius-ulna, biceps brachii muscle-tendon unit, and stabilizers of the shoulder and elbow joints. Specimens were mounted to a rigid board by transfixation pins through the humerus and instrumented for mechanical manipulation of the limb and joint angle and load measurements. Flexion and extension limits of shoulder and elbow joint ranges of motion were measured in each joint separately, while the other joint was fixed. Measurements were made before and after transection of the biceps brachii muscle tendon unit. RESULTS: The biceps brachii muscle-tendon unit limited elbow joint extension when the shoulder joint was fixed in flexion, limited shoulder joint flexion when the elbow joint was fixed in extension, and inhibited shoulder joint extension to a lesser degree when the elbow joint was fixed at midrange angles of 75 degrees to 90 degrees. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical manipulation of the elbow joint into hyperextension during shoulder joint flexion is indicative of biceps brachii injury. PMID- 15822582 TI - Evaluation of developmental changes in the coexpression of myosin heavy chains and metabolic properties of equine skeletal muscle fibers. AB - OBJECTIVE: To determine the growth-related changes in metabolic and anatomic properties in equine muscle fiber type, including hybrid fibers identified with immunohistochemical analysis. ANIMALS: 24 2-, 6-, 12-, and 24-month-old female Thoroughbreds. PROCEDURE: Samples were obtained from the gluteus medius muscle of all horses. Expression of myosin heavy chain (MHC) isoforms MHC-I, -IIa, -IIb, and -IIx in each muscle fiber was detected by use of 4 primary monoclonal antibodies: BA-D5, SC-71, BF-F3, and BF-35, respectively. Five muscle fiber types (types I, I/IIA, IIA, IIA/IIX, and IIX) were immunohistochemically identified. The area and activity of succinic dehydrogenase (SDH) in each fiber type were determined by use of quantitative histochemical staining and image analysis. RESULTS: Although the proportion of type I and IIX fibers did not change with age, the proportion of type IIA and IIA/IIX fibers significantly increased and decreased, respectively, from 2 months to 24 months of age. The increase in proportion of type IIA fibers with growth may have been attributable to muscle fiber-type transition from type IIA/IIX fibers but not from type IIX fibers. Values for SDH activity and fiber area in hybrid fiber types were intermediate to those for their respective pure phenotypes. CONCLUSIONS AND CLINICAL RELEVANCE: Hybrid fibers have an important role for determining the proportion of muscle fiber type in horses < 24 months old, and the metabolic and anatomic properties of the hybrid fibers are well coordinated, as in mature horses. PMID- 15822584 TI - Risk of infection with Cryptosporidium parvum and Cryptosporidium hominis in dairy cattle in the New York City watershed. AB - OBJECTIVE: To determine the risk posed by Cryptosporidium parvum and Cryptosporidium hominis from dairy cattle in the New York City watershed (NYCW). SAMPLE POPULATION: Samples from cattle at risk for shedding Cryptosporidium organisms on randomly selected dairy farms in the NYCW. PROCEDURE: Feces were collected for 4 years from calves at risk for infection on 37 dairies. Oocysts were detected by use of centrifugation concentration-flotation microscopy. The DNA was directly isolated from fecal samples and used to amplify fragments of the small subunit ribosomal RNA and thrombospondin-related adhesion protein C-2 genes by use of nested polymerase chain reaction assays. Small subunit ribosomal RNA fragments were restriction digested by the enzyme Vspl and thrombospondin-related adhesion protein C-2 fragments were digested by Eco91l to distinguish between C hominis (formerly known as genotype 1) and C parvum (formerly known as genotype 2). RESULTS: Of 437 fecal samples examined, 214 contained oocysts. Amplicons were generated for 200 samples. We can be certain, with 95% confidence, that cattle in the NYCW did not harbor C hominis. CONCLUSIONS AND CLINICAL RELEVANCE: Cryptosporidium infections in cattle are under examination because of the potential contamination of public waters by manure. Although cattle may be the source of zoonotic infection via C parvum, they pose little risk for C hominis (the strain commonly isolated from humans in waterborne outbreaks of disease). Other sources of oocysts should be considered when investigating outbreaks attributable to contaminated urban drinking water because cattle pose only a small risk via shedding of C hominis. PMID- 15822583 TI - Influence of medetomidine on stress-related neurohormonal and metabolic effects caused by butorphanol, fentanyl, and ketamine administration in dogs. AB - OBJECTIVE: To examine stress-related neurohormonal and metabolic effects of butorphanol, fentanyl, and ketamine administration alone and in combination with medetomidine in dogs. ANIMALS: 10 Beagles. PROCEDURE: 5 dogs received either butorphanol (0.1 mg/kg), fentanyl (0.01 mg/kg), or ketamine (10 mg/kg) IM in a crossover design. Another 5 dogs received either medetomidine (0.02 mg/kg) and butorphanol (0.1 mg/kg), medetomidine and fentanyl (0.01 mg/kg), medetomidine and ketamine (10 mg/kg), or medetomidine and saline (0.9% NaCI) solution (0.1 mL/kg) in a similar design. Blood samples were obtained for 6 hours following the treatments. Norepinephrine, epinephrine, cortisol, glucose, insulin, and nonesterified fatty acid concentrations were determined in plasma. RESULTS: Administration of butorphanol, fentanyl, and ketamine caused neurohormonal and metabolic changes similar to stress, including increased plasma epinephrine, cortisol, and glucose concentrations. The hyperglycemic effect of butorphanol was not significant. Ketamine caused increased norepinephrine concentration. Epinephrine concentration was correlated with glucose concentration in the butorphanol and fentanyl groups but not in the ketamine groups, suggesting an important difference between the mechanisms of the hyperglycemic effects of these drugs. Medetomidine prevented most of these effects except for hyperglycemia. Plasma glucose concentrations were lower in the combined sedation groups than in the medetomidine-saline solution group. CONCLUSIONS AND CLINICAL RELEVANCE: Opioids or ketamine used alone may cause changes in stress-related biochemical variables in plasma. Medetomidine prevented or blunted these changes. Combined sedation provided better hormonal and metabolic stability than either component alone. We recommend using medetomidine-butorphanol or medetomidine-ketamine combinations for sedation or anesthesia of systemically healthy dogs. PMID- 15822585 TI - Effect of early training on the jumping technique of horses. AB - OBJECTIVE: To investigate the effects of early training for jumping by comparing the jumping technique of horses that had received early training with that of horses raised conventionally. ANIMALS: 40 Dutch Warmblood horses. PROCEDURE: The horses were analyzed kinematically during free jumping at 6 months of age. Subsequently, they were allocated into a control group that was raised conventionally and an experimental group that received 30 months of early training starting at 6 months of age. At 4 years of age, after a period of rest in pasture and a short period of training with a rider, both groups were analyzed kinematically during free jumping. Subsequently, both groups started a 1-year intensive training for jumping, and at 5 years of age, they were again analyzed kinematically during free jumping. In addition, the horses competed in a puissance competition to test maximal performance. RESULTS: Whereas there were no differences in jumping technique between experimental and control horses at 6 months of age, at 4 years, the experimental horses jumped in a more effective manner than the control horses; they raised their center of gravity less yet cleared more fences successfully than the control horses. However, at 5 years of age, these differences were not detected. Furthermore, the experimental horses did not perform better than the control horses in the puissance competition. CONCLUSIONS AND CLINICAL RELEVANCE: Specific training for jumping of horses at an early age is unnecessary because the effects on jumping technique and jumping capacity are not permanent. PMID- 15822586 TI - Effects of ranitidine, famotidine, pantoprazole, and omeprazole on intragastric pH in dogs. AB - OBJECTIVE: To identify the normal gastric acid secretion profile in dogs and determine the degree of gastric acid suppression associated with 4 gastric acid suppressants. ANIMALS: 12 healthy Beagles. PROCEDURE: Intragastric pH was measured continuously for 24-hour periods with a digital recording system placed via a gastrostomy tube. Baseline measurements were obtained when food was withheld and when dogs were fed a standard diet. Dogs were then treated with ranitidine (2 mg/kg, IV, q 12 h), famotidine (0.5 mg/kg, IV, q 12 h), pantoprazole (1 mg/kg, IV, q 24 h), omeprazole (1 mg/kg, PO, q 24 h), or saline solution for 7 days; intragastric pH was recorded on days 0, 2, and 6. Subsequently, the effects of administering famotidine (0.5 mg/kg, IV, q 8 h; 6 dogs) and omeprazole as a suspension (1 mg/kg, PO, q 12 h; 6 dogs) were evaluated. Median 24-hour intragastric pH, percentage of time pH was > or = 3, and percentage of time pH was > or = 4 were determined. RESULTS: Median pH, percentage of time pH was > or = 3, and percentage of time pH was > or = 4 were all significantly higher when food was withheld than when dogs were fed. Famotidine, pantoprazole, and omeprazole significantly suppressed gastric acid secretion, compared with saline solution, as determined on the basis of median 24 hour pH and percentages of time pH was > or = 3 or > or = 4. However, ranitidine did not. Omeprazole suspension suppressed gastric acid secretion. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in healthy dogs, famotidine, pantoprazole, and omeprazole significantly suppress gastric acid secretion. Twice daily administration of a suspension of omeprazole, was the only regimen tested that approached the potential therapeutic efficacy for acid-related disease when assessed by criteria used for human patients. PMID- 15822587 TI - Evaluation of the phospholamban gene in purebred large-breed dogs with dilated cardiomyopathy. AB - OBJECTIVE: To evaluate the role of the phospholamban gene in purebred large-breed dogs with dilated cardiomyopathy (DCM). ANIMALS: 6 dogs with DCM, including 2 Doberman Pinschers, 2 Newfoundlands, and 2 Great Danes. PROCEDURE: All dogs had clinical signs of congestive heart failure, and a diagnosis of DCM was made on the basis of echocardiographic findings. Blood samples were collected from each dog, and genomic DNA was isolated by a salt extraction method. Specific oligonucleotides were designed to amplify the promoter, exon 1, the 5'-part of exon 2 including the complete coding region, and part of intron 1 of the canine phospholamban gene via polymerase chain reaction procedures. These regions were screened for mutations in DNA obtained from the 6 dogs with DCM. RESULTS: No mutations were identified in the promoter, 5' untranslated region, part of intron 1, part of the 3' untranslated region, and the complete coding region of the phospholamban gene in dogs with DCM. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that mutations in the phospholamban gene are not a frequent cause of DCM in Doberman Pinschers, Newfoundlands, and Great Danes. PMID- 15822588 TI - Inheritance of hereditary equine regional dermal asthenia in Quarter Horses. AB - OBJECTIVE: To assess heritability and mode of inheritance for hereditary equine regional dermal asthenia (HERDA) in Quarter Horses. ANIMALS: 1,295 horses with Quarter Horse bloodlines, including 58 horses affected with HERDA. PROCEDURE: Horses were classified as affected or unaffected or as undetermined when data were insufficient to assess phenotype. Pedigree data were analyzed to determine the probable mode of inheritance. Heritability was estimated by use of Bayesian statistical methods. RESULTS: Heritability (mean+/-SD) of HERDA was estimated to be 0.38+/-0.13, with both sexes having an equal probability of being affected. Results for evaluation of the pedigrees were consistent with a single Mendelian autosomal recessive mode of inheritance. CONCLUSIONS AND CLINICAL RELEVANCE: HERDA in Quarter Horses is an inherited disease, and affected horses are more likely to produce affected offspring. An autosomal recessive mode of inheritance should be considered by people making breeding decisions involving Quarter Horses when a first-degree relative has been confirmed with HERDA or has produced affected offspring. In addition, breeders whose horses have produced affected offspring can reduce the likelihood of producing affected horses in the future by avoiding inbreeding. PMID- 15822589 TI - Investigation of the effect of black walnut extract on in vitro ion transport and structure of equine colonic mucosa. AB - OBJECTIVE: To examine the secretory response (in the presence and absence of prostaglandin inhibition) in vitro and structural alterations of colonic mucosa in horses after intragastric administration of black walnut extract (BWE). ANIMALS: 14 adult horses. PROCEDURE: Seven horses were administered BWE intragastrically and monitored for 11 hours. Tissue samples were obtained from the right ventral, left ventral, and right dorsal colons (RVC, LVC, and RDC, respectively) of the 7 BWE-treated and 7 control horses. Tissue samples were examined via light microscopy, and the extent of hemorrhage, edema, and granulocytic cellular infiltration (neutrophils and eosinophils) was graded. Colonic mucosal segments were incubated with or without flunixin meglumine (FLM) for 240 minutes; spontaneous electrical potential difference and short-circuit current (Isc) were recorded and used to calculate mucosal resistance. RESULTS: Colonic tissues from BWE-treated horses (with or without FLM exposure) had an overall greater Isc during the 240-minute incubation period, compared with tissues from control horses. The resistance pattern in RVC, LVC, and RDC samples (with or without FLM exposure) from BWE-treated horses was decreased overall, compared with control tissues (with or without FLM exposure). Histologically, colonic mucosal tissues from BWE-treated horses had more severe inflammation (involving primarily eosinophils), edema, and hemorrhage, compared with tissue from control horses. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, BWE administration appears to cause an inflammatory response in colonic mucosal epithelium that results in mucosal barrier compromise as indicated by decreased mucosal resistance with presumed concomitant electrogenic chloride secretory response, which is not associated with prostaglandin mediation. PMID- 15822590 TI - Expression of erythropoietin in cats treated with a recombinant adeno-associated viral vector. AB - OBJECTIVE: To characterize the biological effects of IM administration of a recombinant adeno-associated virus serotype 2 (rAAV2) vector containing feline erythropoietin (fEPO) cDNA and determine whether readministration of the vector or removal of muscle tissue at the injection sites alters those effects. ANIMALS: 10 healthy 7-week-old specific pathogen-free cats. PROCEDURE: Cats received 1 X 10(7) infective units (iU; n = 3), 1 X 10(8) iU (3), or 1 X 10(9) iU (2) of rAAV2 fEPO vector IM (day 0). Two control cats received an rAAV2 vector containing the LacZ gene (1 X 10(9) iU, IM). In all cats, hematologic variables and serum fEPO concentration were measured at intervals; anti-rAAV2 antibody titer was measured on day 227. In cats that did not respond to treatment, the rAAV2-fEPO vector was readministered. Injection sites were subsequently surgically removed. RESULTS: Compared with control cats, cats treated with 1 X 10(9) iU of rAAV2-fEPO vector had increased Hct and serum fEPO concentrations. One of these cats developed pure RBC aplasia; its Hct normalized following injection site excision. Cats receiving lower doses of vector had no response; on retreatment, 1 of those cats developed sustained erythrocytosis that persisted despite injection site removal and the others did not respond or responded transiently. Antibodies against rAAV2 were detected in all vector-treated cats. CONCLUSIONS AND CLINICAL RELEVANCE: Gene therapy may be an effective treatment for cats with hypoproliferative anemia. However, rAAV2-fEPO vector administration may result in pure RBC aplasia or pathologic erythrocytosis, and injection site removal does not consistently abolish the biological response. PMID- 15822591 TI - Association of maximum voluntary dietary intake of freeze-dried garlic with Heinz body anemia in horses. AB - OBJECTIVE: To characterize hematologic and clinical consequences of chronic dietary consumption of freeze-dried garlic at maximum voluntary intake in horses. ANIMALS: 4 healthy sex- and age-matched horses. PROCEDURE: An initial garlic dose (0.05 g/kg, twice daily) was fed to 2 horses in a molasses carrier as part of their normal ration and was gradually increased to maximum voluntary intake (0.25 g/kg, twice daily) over 41 days. Dietary supplementation then continued for a total of 71 days. Two control horses were fed molasses with no garlic with their ration. Blood samples were collected weekly and analyzed for hematologic and biochemical changes, including the presence of Heinz bodies. Recovery of affected blood values was followed for 5 weeks after termination of dietary supplementation with garlic. RESULTS: At a daily dose of > 0.2 g/kg, horses fed garlic developed hematologic and biochemical indications of Heinz body anemia, as characterized by increases in Heinz body score (HBS), mean corpuscular volume (MCV), mean corpuscular hemoglobin, platelet count, and serum unconjugated and total bilirubin concentrations and decreases in RBC count, blood hemoglobin concentration, mean corpuscular hemoglobin concentration, and serum haptoglobin concentration. Recovery from anemia was largely complete within 5 weeks after termination of dietary supplementation with garlic. Heinz body score and MCV remained high at the end of the 5-week recovery period. CONCLUSIONS AND CLINICAL RELEVANCE: Horses will voluntarily consume sufficient quantities of garlic to cause Heinz body anemia. The potential for garlic toxicosis exists when horses are chronically fed garlic. Further study is required to determine the safe dietary dose of garlic in horses. PMID- 15822592 TI - Effects of oral potassium supplementation on acid-base status and plasma ion concentrations of horses during endurance exercise. AB - OBJECTIVE: To compare effects of oral supplementation with an experimental potassium-free sodium-abundant electrolyte mixture (EM-K) with that of oral supplementation with commercial potassium-rich mixtures (EM+K) on acid-base status and plasma ion concentrations in horses during an 80-km endurance ride. ANIMALS: 46 healthy horses. PROCEDURE: Blood samples were collected before the ride; at 21-, 37-, 56-, and 80-km inspection points; and during recovery (ie, 30 minute period after the ride). Consumed electrolytes were recorded. Blood was analyzed for pH, PvCO2, and Hct, and plasma was analyzed for Na+, K+, Cl-, Ca2+, Mg2+, lactate, albumin, phosphate, and total protein concentrations. Plasma concentrations of H+ and HCO3-, the strong ion difference (SID), and osmolarity were calculated. RESULTS: 34 (17 EM-K and 17 EM+K treated) horses finished the ride. Potassium intake was 33 g less and Na+ intake was 36 g greater for EM-K treated horses, compared with EM+K-treated horses. With increasing distance, plasma osmolarity; H+, Na+, K+, Mg2+, phosphate, lactate, total protein, and albumin concentrations; and PvCO2 and Hct were increased in all horses. Plasma HCO3-, Ca2+, and Cl- concentrations were decreased. Plasma H+ concentration was significantly lower in EM-K-treated horses, compared with EM+K-treated horses. Plasma K+ concentrations at the 80-km inspection point and during recovery were significantly less in EM-K-treated horses, compared with EM+K-treated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in plasma H+ and K+ concentrations in this endurance ride were moderate and unlikely to contribute to signs of muscle fatigue and hyperexcitability in horses. PMID- 15822593 TI - Investigation of antigen-specific T-cell responses and subcutaneous granuloma development during experimental sensitization of calves with Mycobacterium avium subsp paratuberculosis. AB - OBJECTIVE: To characterize the early cellular immune response to Mycobacterium avium subsp paratuberculosis (MAP) infection and evaluate the development of granulomatous inflammation at the SC injection site in experimentally inoculated calves. ANIMALS: Forty-eight 4-week-old calves. PROCEDURE: Calves received an SC injection of MAP strain 19698 (n = 25), sterile saline (0.9% NaCl) solution (20), or a commercial paratuberculosis vaccine (3); the inoculation site tissue and associated draining lymph node were excised at postinoculation day (PID) 0 (n = 36), 7 (14), 14 (6), 21 (8), and 60 (32). Sections of inoculation site tissues were evaluated immunohistochemically for T-cell subsets; lymph node mononuclear cells (LNMCs) were assessed for T-cell surface markers and for intracellular interferon-gamma via flow cytometry. RESULTS: At MAP inoculation sites, calves developed mild, focal granulomatous inflammation by PID 7; by PID 60, areas of inflammation contained macrophages with numerous lymphocytes. Compared with control calves, there was increased antigen-specific LNMC proliferation in MAP- and vaccine-inoculated calves at PID 60, although proliferation among lymphocyte subsets was not significantly different between MAP-inoculated and control calves; in vaccine-inoculated calves, CD4+ T-cells predominated. In MAP inoculated and control calves, antigen-specific interferon-gamma production by LNMCs did not differ significantly; vaccine-inoculated calves had marked interferon-gamma expression by CD4+ T-cells. CONCLUSIONS AND CLINICAL RELEVANCE: In calves, SC administration of MAP resulted in granulomatous inflammation at inoculation sites and an antigen-specific T-cell proliferative response. Results suggest that this experimental system can be used to reproducibly generate antigen-specific T-cells during MAP infection for functional analysis. PMID- 15822594 TI - Effects of CD28 blockade on subsets of naive T cells in cats. AB - OBJECTIVE: To determine whether human CTLA4-Ig ([hu]CTLA4-Ig) inhibits costimulation-dependent lymphocyte proliferation in vitro, compare the effects of (hu)CTLA4-Ig with cyclosporine and steroids on CD4+ and CD8+ T-cell lymphocyte proliferation, and determine whether memory T-cell function remains intact in the presence of (hu)CTLA4-Ig. ANIMALS: 29 cats. PROCEDURE: Peripheral blood mononuclear cells (PBMCs) were stimulated with concanavalin A (costimulation dependent mitogen) or phorbol 12-myristate 13-acetate and ionomycin (costimulation independent mitogens) alone or in the presence of (hu)CTLA4-Ig, cyclosporine, or dexamethasone; effects of these treatments on lymphocyte proliferation were assessed by incorporation of thymidine labeled with tritium or flow cytometry. Antigen-specific proliferation was determined by stimulating PBMCs from 2 healthy cats seropositive for Toxoplasma gondii with soluble Toxoplasma antigen alone or in the presence of (hu)CTLA4-Ig or cyclosporine. RESULTS: (hu)CTLA4-Ig inhibited costimulation-dependent lymphocyte proliferation in vitro but had no effect on costimulation-independent lymphocyte proliferation. Compared with mitogen alone, (hu)CTLA4-Ig caused a significant decrease in responder frequency and proliferative capacity of CD4+ T cells; however, the effect on CD8+ T cells was not significant. Cyclosporine alone or with dexamethasone had a significantly greater suppressive effect on responder frequency and proliferative capacity of CD4+ and CD8+ T cells, compared with (hu)CTLA4-Ig. Compared with cyclosporine, (hu)CTLA4-Ig appeared to have a sparing effect on antigen-specific proliferation of memory CD4+ and CD8+ T cells. CONCLUSIONS AND CLINICAL RELEVANCE: (hu)CTLA4-Ig selectively inhibited costimulation-dependent proliferation of lymphocytes in vitro and had a sparing effect on antigen-specific proliferation of memory cells. The specificity of its mechanism of action suggests that (hu)CTLA4-Ig may prevent allograft rejection but leave memory responses to previously encountered antigens intact. PMID- 15822595 TI - Detection of synovial macrophages in the joint capsule of dogs with naturally occurring rupture of the cranial cruciate ligament. AB - OBJECTIVE: To test the hypotheses that the densities of macrophages in the synovial membranes and capsules of stifle joints in dogs with ruptured cranial cruciate ligaments are greater than those of normal joints and that those densities in affected joints are positively correlated with the chronicity and severity of the disease. ANIMALS: 17 dogs with naturally occurring rupture of the cranial cruciate ligament and 5 healthy control dogs. PROCEDURE: All dogs underwent orthopedic and radiographic evaluations. In affected dogs, duration of clinical signs was used as an indicator of disease chronicity and the severity of osteoarthritis in the stifle joint was determined radiographically. Joint capsule specimens were evaluated histologically; macrophages, interleukin-6, and tumor necrosis factor-alpha were identified by use of immunocytochemical techniques. RESULTS: Compared with unaffected joints, macrophage density was increased in all affected joints. Duration of disease was significantly associated with radiographic severity of osteoarthritis and synovial macrophage density. Synovial macrophage density was significantly associated with severity of osteoarthritis and with the presence of interleukin-6 and tumor necrosis factor-a. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that synovial macrophages may be involved in the development of pathologic changes (including osteophyte formation) in the stifle joints of dogs with osteoarthritis secondary to rupture of the cranial cruciate ligament. Determination of the importance of synovial macrophages in the development of changes in osteoarthritic joints may result in new treatment strategies that involve elimination of the deleterious effects of those cells. PMID- 15822596 TI - Evaluation of the effect of routine histologic processing on the size of skin samples obtained from dogs. AB - OBJECTIVE: To determine the effects that routine histologic processing has on the dimensions of samples of normal skin of dogs and assess whether the inclusion of a muscle or fascial layer in such samples alters those effects. SAMPLE POPULATION: Skin samples obtained from 6 medium-sized adult dogs with grossly normal skin. PROCEDURE: From each dog, skin samples (with or without underlying fascia or muscle) were obtained from 3 sites bilaterally (6 samples/dog) and processed routinely for histologic evaluation; their dimensions were measured at intervals during the experiment. RESULTS: As a result of processing, skin samples decreased in size (combined percentage change in length and width) and increased in thickness, compared with their original dimensions. Samples without fascia or muscle decreased in size by 21.1% to 32.0% and increased in thickness by 45.1 % to 75.8%. The site of sample origin influenced processing-associated changes in sample size but did not affect the change in thickness. Decreases in dimensions did not vary with inclusion of fascia but did vary with inclusion of muscle. The change in thickness did not vary with inclusion of a layer of fascia or muscle. CONCLUSIONS AND CLINICAL RELEVANCE: Processing of skin samples obtained from dogs for histologic evaluation can cause changes in sample dimensions; samples may decrease in length and width by as much as 32% and increase in thickness by 75.8%, compared with their original dimensions. The presence of muscle in canine skin samples can restrict the amount of shrinkage in length or width associated with processing. PMID- 15822597 TI - Investigation of the induction of antibodies against Crandell-Rees feline kidney cell lysates and feline renal cell lysates after parenteral administration of vaccines against feline viral rhinotracheitis, calicivirus, and panleukopenia in cats. AB - OBJECTIVE: To determine whether administration of Crandell-Rees feline kidney (CRFK) cell lysates or vaccines against feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP vaccines) that likely contain CRFK cell proteins induces antibodies against CRFK cell or feline renal cell (FRC) lysates in cats. ANIMALS: 14 eight-week-old cats. PROCEDURE: Before and after the study, renal biopsy specimens were obtained from each cat for histologic evaluation. Each of 4 FVRCP vaccines was administered to 2 cats at weeks 0, 3, 6, and 50. Between weeks 0 and 50, another 3 pairs of cats received 11 CRFK cell lysate inoculations SC (10, 50, or 50 microg mixed with alum). Clinicopathologic evaluations and ELISAs to detect serum antibodies against CRFK cell or FRC lysates were performed at intervals. RESULTS: Cats had no antibodies against CRFK cell or FRC lysates initially. All cats administered CRFK cell lysate had detectable antibodies against CRFK cell or FRC lysates on multiple occasions. Of 6 cats vaccinated parenterally, 5 had detectable antibodies against CRFK cell lysate at least once, but all 6 had detectable antibodies against FRC lysate on multiple occasions. Cats administered an intranasal-intraocular vaccine did not develop detectable antibodies against either lysate. Important clinicopathologic or histologic abnormalities were not detected during the study. CONCLUSIONS AND CLINICAL RELEVANCE: Parenteral administration of vaccines containing viruses likely grown on CRFK cells induced antibodies against CRFK cell and FRC lysates in cats. Hypersensitization with CRFK cell proteins did not result in renal disease in cats during the 56-week study. PMID- 15822598 TI - Computed tomography and cross-sectional anatomy of the thorax in clinically normal dogs. AB - OBJECTIVE: To provide a detailed anatomic description of the thorax in clinically normal dogs by means of computed tomography. ANIMALS: 4 clinically normal adult German Shepherd Dogs weighing 28 to 37 kg. PROCEDURE: Dogs were anesthetized and positioned in ventral recumbency for computed tomographic (CT) examination of the thorax. A CT image from the thoracic inlet to the diaphragm was made by use of a third-generation scanner with a slice thickness of 5 mm. Individual images were reviewed by use of soft tissue (window width, 250 Hounsfield units; window level, 35 Hounsfield units) and lung (window width, 1,000 Hounsfield units; window level, -690 Hounsfield units) settings. One dog, weighing 28 kg, was euthanatized, bound on a wooden frame in the same position as used for CT examination, and frozen at -14 degrees C until solid. By use of an electric band saw, the frozen thorax was sectioned at 10-mm-thick intervals. Slab sections were immediately cleaned, photographed, and compared with corresponding CT images. RESULTS: Anatomic sections were studied, and identified anatomic structures were matched with structures on corresponding CT images. Except for some blood vessels and details of the heart, most of the bony and soft tissue structures of the thorax discerned on anatomic slices could be found on matched CT images. CONCLUSIONS AND CLINICAL RELEVANCE: Because CT images provide detailed information on most structures of the canine thorax, results of our study could be used as a guide for evaluation of CT images of the thorax of dogs with thoracic diseases. PMID- 15822599 TI - Effects of Carolina rinse solution, dimethyl sulfoxide, and the 21-aminosteroid, U-74389G, on microvascular permeability and morphology of the equine jejunum after low-flow ischemia and reperfusion. AB - OBJECTIVE: To evaluate effects of Carolina rinse solution, dimethyl sulfoxide (DMSO), and 21-aminosteroid, U-74389G, on microvascular permeability and morphology of the equine jejunum after low-flow ischemia and reperfusion. ANIMALS: 20 healthy adult horses. PROCEDURE: Under anesthesia, full-thickness biopsy specimens of a distal portion of the jejunum were obtained for baseline measurements. In addition to a control segment, 2 jejunal segments were identified as sham-operated or experimental segments. Experimental segments underwent 60 minutes of low-flow ischemia and 3.5 hours of reperfusion. Treatments were as follows: U-74389G (3 mg/kg, IV; 6 horses), DMSO (20 mg/kg, IV; 6) diluted in 1 L of saline (0.9% NaCl) solution, local perfusion (via jejunal artery) of Carolina rinse solution (0.5 mL/kg; 4), and local perfusion of lactated Ringer's solution (0.5 mL/kg; 4). RESULTS: Jejunal microvascular permeability was significantly lower after treatment with Carolina rinse solution or DMSO, compared with U-74389G or lactated Ringer's solution treatments. After DMSO treatment, serosal- and submucosal-layer edema was significantly increased in experimental segments, compared with control or sham-operated segments; however, edema increases were significantly less than for lactated Ringer's solution or U-74389G treatments. Significant decreases in intestinal wet weight to-dry weight ratio were found following Carolina rinse solution or DMSO treatments, compared with lactated Ringer's solution or U-74389G treatments. Edema formation and leukocyte infiltration in jejunal segments of horses treated with lactated Ringer's solution or U-74389G were increased, compared with Carolina rinse solution or DMSO treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Carolina rinse solution and DMSO may be protective against ischemia-reperfusion injury in the equine jejunum. PMID- 15822600 TI - Ultrasonographic measurement of abomasal volume, location, and emptying rate in calves. AB - OBJECTIVE: To develop and validate an ultrasonographic method for measuring abomasal volume, location, and emptying rate in suckling calves. ANIMALS: 9 male Holstein calves < 40 days of age. PROCEDURE: Before and after calves were fed different volumes of milk replacer or 2 L of oral electrolyte solutions, ultrasonographic measurements of abomasal dimensions (width, length, and height) were obtained by applying a 3.5-MHz sector probe to the ventral aspect of the abdomen in the transverse and sagittal planes. Abomasal volume was calculated from the ultrasonographic measurements by modeling the abomasum as an ellipsoid and by use of a power exponential equation to calculate the half-time of abomasal emptying (t1/2). RESULTS: Preprandial abomasal volume was 20 to 137 mL. All 3 abomasal dimensions increased during feeding and after suckling, and the abomasum was symmetrically located about the midline of the ventral aspect of the abdomen. Strong linear relationships were identified between ultrasonographic and suckled volumes, between ultrasonographic and scintigraphic heights, and between ultrasonographic and scintigraphic lengths. Ultrasonographic t1/2 was linearly related to scintigraphic t1/2; the latter is regarded as the gold standard measure of gastric emptying rate. CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasonographic evaluation of the abomasum appears to provide a practical, rapid, noninvasive, and accurate method for determination of abomasal volume, location, and emptying rate in suckling calves. PMID- 15822601 TI - Assessment of the effects of erythromycin, neostigmine, and metoclopramide on abomasal motility and emptying rate in calves. AB - OBJECTIVE: To determine and compare the effects of erythromycin, neostigmine, and metoclopramide on abomasal motility and emptying rate in suckling calves. ANIMALS: 6 male Holstein calves (15 to 40 days of age). PROCEDURE: Calves were monitored for 1 hour before being fed milk replacer (60 mL/kg; time, 0 minutes) and then were monitored for another 3 hours. Calves received 6 treatments in randomized order: erythromycin (8.8 mg/kg, IM) at -30 minutes; low-dose erythromycin (0.88 mg/kg, IM) at -30 minutes; erythromycin (8.8 mg/kg, IM) at -30 minutes and neostigmine (0.02 mg/kg, SC) at -30 and 90 minutes; neostigmine (0.02 mg/kg, SC) at -30 and 90 minutes; metoclopramide (0.1 mg/kg, IM) at-30 and 90 minutes; and placebo (2 mL of saline [0.9% NaCl] solution, SC) at -30 minutes. Abomasal volume was calculated from ultrasonographic measurements of abomasal width, length, and height. Abomasal motility and emptying rate were assessed by measuring luminal pressure and change in abomasal volume over time. RESULTS: Administration of erythromycin (8.8 mg/kg) increased the frequency of abomasal luminal pressure waves and the mean abomasal luminal pressure and decreased the half-time of abomasal emptying by 37%. Administration of metoclopramide, neostigmine, and low-dose erythromycin (0.88 mg/kg) did not alter abomasal motility, mean luminal pressure, or emptying rate. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that administration of erythromycin at the labeled antimicrobial dose (8.8 mg/kg, IM) exerted an immediate, marked prokinetic effect in healthy suckling calves, whereas administration of metoclopramide or neostigmine did not alter abomasal motility or emptying rate. PMID- 15822602 TI - Individual organisms as units of analysis: Bayesian-clustering alternatives in population genetics. AB - Population genetic analyses traditionally focus on the frequencies of alleles or genotypes in 'populations' that are delimited a priori. However, there are potential drawbacks of amalgamating genetic data into such composite attributes of assemblages of specimens: genetic information on individual specimens is lost or submerged as an inherent part of the analysis. A potential also exists for circular reasoning when a population's initial identification and subsequent genetic characterization are coupled. In principle, these problems are circumvented by some newer methods of population identification and individual assignment based on statistical clustering of specimen genotypes. Here we evaluate a recent method in this genre--Bayesian clustering--using four genotypic data sets involving different types of molecular markers in non-model organisms from nature. As expected, measures of population genetic structure (F(ST) and phiST) tended to be significantly greater in Bayesian a posteriori data treatments than in analyses where populations were delimited a priori. In the four biological contexts examined, which involved both geographic population structures and hybrid zones, Bayesian clustering was able to recover differentiated populations, and Bayesian assignments were able to identify likely population sources of specific individuals. PMID- 15822603 TI - Limitations of a two-step moment method for mapping linked multiple QTL. AB - As an alternative to multiple-interval mapping a two-step moment method was recently proposed to map linked multiple quantitative trait loci (QTLs). The advantage of this moment method was supposed to be its simplicity and computational efficiency, especially in detecting closely linked QTLs within a marker bracket, but also in mapping QTLs in different marker intervals. Using simulations it is shown that the two-step moment method may give poor results compared with multiple-interval mapping, irrespective of whether the QTLs are in the same or in different marker intervals, especially if linked QTLs are in repulsion. The criteria of comparison are number of identified QTLs, likelihood ratio test statistics, means and empirical standard errors of the QTL position and QTL effects estimates, and the accuracy of the residual variance estimates. Further, the joint conditional probabilities of QTL genotypes for two putative QTLs within a marker interval were derived and compared with the unmodified approach ignoring the non-independence of the conditional probabilities. PMID- 15822604 TI - Identification of a cryptic lethal mutation in the mouse t(w73) haplotype. AB - t haplotypes are naturally occurring, variant forms of the t complex on mouse chromosome 17, characterized by the presence of four inversions with respect to wild-type. They harbour mutations causing male sterility, male transmission ratio distortion (TRD) and embryonic lethality. Mice carrying t haplotypes have been found throughout the world, and genetic studies of the lethal mutations have identified at least 16 complementation groups. The embryonic lethal phenotypes of many t haplotypes have been characterized in detail, and are thought to be the consequence of homozygosity for single gene mutations. However, the existence of additional mutations in genes that function at later stages of development would be obscured. Here we investigated the possibility of multiple mutations in t haplotypes by screening the t(w73) haplotype for the presence of novel mutations. Since genetic analysis of t haplotype mutations is hindered by recombination suppression due to the inversions, deletion complexes covering the proximal two thirds of the t complex were used to uncover the presence of any new lethal alleles. This analysis revealed a novel mutation between D17Jcs41 and D17Mit100, causing mice carrying both t(w73) and selected deletions to die at birth, prior to feeding. The finding of a new, cryptic lethal mutation in t haplotypes is an indication that these recombinationally isolated chromosomes, which already contain at least one lethal mutation that prevents homozygosity, may serve as sinks for the accumulation of additional recessive mutations. PMID- 15822605 TI - Marker-assisted introgression of the Compact mutant myostatin allele MstnCmpt dl1Abc into a mouse line with extreme growth effects on body composition and muscularity. AB - Myostatin is a negative regulator of muscle growth and mutations in its gene lead to muscular hypertrophy and reduced fat. In cattle, this is seen in 'double muscled' breeds. We have used marker-assisted introgression to introduce a murine myostatin mutation, MstnCmpt-dl1Abc [Compact (C)], into an inbred line of mice (DUHi) that had been selected on body weight and had exceptional growth. Compared with homozygous wild-type mice, homozygous (C/C) mice of this line were approximately 4-5 % lighter, had approximately 7-8 % shorter tails, substantially increased muscle weights (e.g. quadriceps muscle in males was 59 % heavier) and an increased 'dressing percentage' (approximately 49 % vs 39 %), an indicator of overall muscularity. The weights of several organs (e.g. liver, kidney, heart and digestive tract) were significantly reduced, by 12-20 %. Myostatin deficiency also resulted in drastic reductions of total body fat and of various fat depots, total body fat proportion falling from approximately 17.5 % in wild-type animals of both sexes to 9.5 % and 11.6% in homozygous (C/C) females and males, respectively. Males with a deficiency in myostatin had higher gains in muscle traits than females. Additionally, there was a strong distortion of the segregation ratio on the DUHi background. Of 838 genotyped pups from inter se matings 29 %, 63 % and 8 % were homozygous wild type (+/+), heterozygous (C/+) and homozygous (C/C), respectively, showing that MstnCmpt-dl1Abc has lower fitness on this background. This line, when congenic, will be a useful resource in gene expression studies and for finding modifying genes. PMID- 15822606 TI - Hitchhiking and recombination in birds: evidence from Mhc-linked and unlinked loci in Red-winged Blackbirds (Agelaius phoeniceus). AB - Hitchhiking phenomena and genetic recombination have important consequences for a variety of fields for which birds are model species, yet we know virtually nothing about naturally occurring rates of recombination or the extent of linkage disequilibrium in birds. We took advantage of a previously sequenced cosmid clone from Red-winged Blackbirds (Agelaius phoeniceus) bearing a highly polymorphic Mhc class II gene, Agph-DABI, to measure the extent of linkage disequilibrium across approximately 40 kb of genomic DNA and to determine whether non-coding nucleotide diversity was elevated as a result of physical proximity to a target of balancing selection. Application of coalescent theory predicts that the hitchhiking effect is enhanced by the larger effective population size of blackbirds compared with humans, despite the presumably higher rates of recombination in birds. We surveyed sequence polymorphism at three Mhc-linked loci occurring 1.5-40 kb away from Agph-DAB1 and found that nucleotide diversity was indistinguishable from that found at three presumably unlinked, non-coding introns (beta-actin intron 2, beta-fibrinogen intron 7 and rhodopsin intron 2). Linkage disequilibrium as measured by Lewontin's D' was found only across a few hundred base pairs within any given locus, and was not detectable among any Mhc-linked loci. Estimated rates of the per site recombination rate p derived from three different analytical methods suggest that the amounts of recombination in blackbirds are up to two orders of magnitude higher than in humans, a discrepancy that cannot be explained entirely by the higher effective population size of blackbirds relative to humans. In addition, the ratio of the number of estimated recombination events per mutation frequently exceeds 1, as in Drosophila, again much higher than estimates in humans. Although the confidence limits of the blackbird estimates themselves span an order of magnitude, these data suggest that in blackbirds the hitchhiking effect for this region is negligible and may imply that the per site per individual recombination rate is high, resembling those of Drosophila more than those of humans. PMID- 15822607 TI - Detecting event-based prospective memory cues occurring within and outside the focus of attention. AB - Event-based prospective memory cues are environmental stimuli that are associated with a previously established intention to perform an activity. Such cues traditionally have been placed in materials that receive focal attention during an ongoing activity. This article reports a direct comparison of event-based cues that occurred either within the focus of attention or at the periphery of such attention. When the cue occurred outside focal attention, manipulating that cue changed event-based prospective memory. The identical manipulation had no effect on event-based responding if the cue occurred within focal attention. These results suggest that cue characteristics can compensate for attention being directed away from an aspect of an ongoing task that contains event-based prospective memory. PMID- 15822608 TI - Priming and false memories from Deese-Roediger-McDermott lists on a fragment completion test with children. AB - This study examined priming and false memories with children on a word fragment completion task using the Deese-Roediger-McDermott (DRM) paradigm. Forty-five 4th and 5th-grade children were shown lists of words and instructed to fill in fragments with the first word that came to mind (implicit instructions) or with the words that were presented during study (explicit instructions). Reliable priming to critical lure words was found under implicit retrieval instructions, and false memory to critical lure words was found under explicit retrieval instructions. However, priming under implicit retrieval instructions did not depend on whether the critical lure word was in the study list. In addition, greater false memory was observed under explicit test instructions. The results replicate and extend research on DRM false memory illusion with children to include implicit retrieval and word fragment completion. Explanations of false memory including gist failure (Brainerd, Reyna, & Forrest, 2002) and implicit associative response (Underwood, 1965) are considered. PMID- 15822609 TI - A psychometric analysis of chess expertise. AB - This study introduces the Amsterdam Chess Test (ACT). The ACT measures chess playing proficiency through 5 tasks: a choose-a-move task (comprising two parallel tests), a motivation questionnaire, a predict-a-move task, a verbal knowledge questionnaire, and a recall task. The validity of these tasks was established using external criteria based on the Elo chess rating system. Results from a representative sample of active chess players showed that the ACT is a very reliable test for chess expertise and that ACT has high predictive validity. Several hypotheses about the relationships between chess expertise, chess knowledge, motivation, and memory were tested. Incorporating response latencies in test scores is shown to lead to an increase in criterion validity, particularly for easy items. PMID- 15822610 TI - Bimodal format effects in working memory. AB - This work combines presentation formats to test whether bimodal conditions offer advantages or disadvantages relative to single formats in working memory performance. A dual task that included recall of 3 or 6 items while verifying the accuracy of math sentences was used in 2 experiments. When comparisons were made between single- and dual-format conditions, there was an advantage for items presented as spoken words and pictures simultaneously and individually. In Experiment 2, dual-format conditions had incongruent information, and spoken words were found to interfere with recall of long sequences of pictures and printed words. The findings suggest that when dual-format items are the same, there are some performance advantages when spoken words are combined with pictures or printed words. When the dual formats are displaying different items, however, spoken words are a more powerful distractor than pictures or printed words, and verbal and visual short-term stores can demonstrate similar susceptibility to distractor interference. PMID- 15822611 TI - Comparison of the efficacy of two name-learning techniques: expanding rehearsal and name-face imagery. AB - Two name-learning techniques were compared: expanding rehearsal and name-face imagery. Participants studied name-face associations and were given a cued recall test in which they were presented with a face and were to recall the name. They were presented with either an expanding rehearsal schedule (expanding condition), a distinctive facial feature coupled with a word phonologically similar to the last name and an interactive image linking the name and facial feature (name-face imagery condition), or a no memory (control) strategy. The expanding rehearsal schedule led to superior name learning relative to the name-face imagery and control conditions after a 15-min (Experiment 1) or 48-hr (Experiment 2) retention interval. In Experiment 3, the retrieval practice explanation was tested but not supported; we argue that an encoding variability interpretation is consistent with the overall pattern of results. Applied implications are also discussed. PMID- 15822612 TI - Dissociation, cognitive failures, and working memory. AB - The relationships between two personality measures (dissociation and cognitive failures) and different measures of interference from secondary tasks for working memory are investigated. Although the personality and cognitive measures were correlated with one another, canonical and bivariate correlations revealed no systematic relationships between the personality measures and the working memory measures. The upper bounds of the confidence intervals showed that, at most, only 10% of the variance was shared. We discuss the implications for theories of dissociation and cognitive functioning. PMID- 15822613 TI - False memories of having said the unsaid: on the importance of a prior intention to speak. AB - Previous research found that adults who participated in a mock debate often later reported having made a point that they had not. In an extension of that study, the present research not only replicated that finding but also investigated the possibility that such errors were the result of the subjects having been induced into intending to make the unmade point. Specifically, the present results show that the occurrence of the inducing stimulus probably did produce such intentions and that the stimulus was also important to the production of subsequent memory errors. Finally, it is argued that the latter errors are most parsimoniously understood as being caused by distorted memories of the intentions demonstrated by the former result. PMID- 15822614 TI - The experimental psychology of attitude change and the tradition of classical rhetoric. AB - Social psychologists might be surprised to learn that their discipline has been cut off from a vast and ancient family tree. The study of attitude change in the context of experimental social psychology began around 1918. It developed into a defined discipline in the 1930s and 1940s, particularly through the work of Carl Hovland and his associates. Unlike earlier specialties in experimental psychology, social psychology emerged well after the 19th-century split between psychology and philosophy in college curricula. Before this period of growth in empirical teaching and practice, the study of persuasion in classical rhetoric was a bedrock of higher education for more than 2000 years. Because of social psychology's late development in empirical science, there is a historical disconnect between experimental social psychology and its ancient philosophical counterpart, classical rhetoric. This article demonstrates similarities and differences between Hovland's findings and the theoretical groundings of classical rhetoric. We suggest areas where modern social psychology might benefit from a look at the older, more holistic theories of the art of rhetoric. PMID- 15822615 TI - Synthesis of (+/-)-4'-ethynyl and 4'-cyano carbocyclic analogues of stavudine (d4T). AB - The synthesis of (+/-)-4'-ethynyl (8) and 4'-cyano (9) carbocyclic analogues of the anti-HIV agent stavudine (5, d4T) is reported. The carbocyclic unit (16) was constructed from readily available beta-keto ester 10. The ethynyl or cyano group of 8 and 9 were prepared, after the introduction of thymine base to 16, by manipulation of the ester function. Evaluation of the anti-HIV activity of 8 and 9 was also carried out. PMID- 15822616 TI - Synthesis of two 8-[(anthraquinone-2-yl)-linked]-2'-deoxyadenosine 3'-benzyl hydrogen phosphates for studies of photoinduced hole transport in DNA. AB - The challenge in working with anthraquinone-2'-deoxyadenosine (AQ-dA) conjugates is that they are insoluble in water and only sparingly soluble in most organic solvents. However, water-soluble AQ-dA conjugates with short linkers are required for study of their electrochemical and intramolecular electron transfer properties in this solvent prior to their use in laser kinetics investigations of photoinduced hole (cation) transport in DNA. This article first describes the synthesis of a water-soluble, ethynyl-linked AQ-dA conjugate, 8-[(anthraquinone-2 yl)ethynyl]-2'-deoxyadenosine 3'-benzyl hydrogen phosphate, based on initial formation of a 5'-O-(4,4'-dimethoxytrityl) (5'-O-DMTr) intermediate. Because intended H2 over Pd/C reduction of the ethynyl linker in 5'-O-DMTr-protected 2' deoxyadenosines cleaves the DMTr protecting group and precipitates multiple side products, this work also describes the synthesis of an ethylenyl-linked AQ-dA conjugate, 8-[2-(anthraquinone-2-yl)ethyl]-2'-deoxyadenosine 3'-benzyl hydrogen phosphate, starting with a 5'-O-tert-butyldiphenylsilyl protecting group. PMID- 15822617 TI - Synthesis of some novel N-ribosyl-1,2,4-triazin-6(1H)-/ones or thiones as potential antibacterial and antifungal chemotherapeutics. AB - Ribosylation of 3-aryl-5-benzyl(or substituted benzyl)-1,2,4-triazin-6(1H)-/ones or thiones with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose gave the corresponding 3-aryl-5-benzyl(or substituted benzyl)-1-(2,3,5-tri-O-benzoyl-beta D-ribofuranosyl)-1,2,4-triazin-6(1H)-/ones or thiones. The structure of the new ribosides was confirmed chemically and spectroscopically. PMID- 15822618 TI - Novel approach to nucleoside-5'-(1-hydroxymethylene-1, 1-bisphosphonates): synthesis of new AZT analogues. AB - An efficient synthetic method of nucleoside-5'-(1-hydroxymethylene-1,1 bisphosphonates) is reported here. The procedure was optimized with 3'-protected thymidine and then applied to synthesis of new AZT analogues. PMID- 15822619 TI - Synthesis and biological evaluation of 2',3'-didehydro-2',3'-dideoxy-9 deazaguanosine, a monophosphate prodrug and two analogues, 2',3'-dideoxy-9 deazaguanosine and 2',3'-didehydro-2',3'-dideoxy-9-deazainosine. AB - 2',3'-Didehydro-2',3'-dideoxy-9-deazaguanosine (1), its monophosphate prodrug (2), and two analogues, 2',3'-dideoxy-9-deazaguanosine (3) and 2',3'-didehydro 2',3'-dideoxy-9-deazainosine (4), have been synthesized from benzoylated 9 deazaguanosine (5). Basic hydrolysis of 5, selective protection of the 2-amino and 5'-hydroxy functions with isobutyryl and silyl groups, respectively, followed by reaction with thiocarbonyldiimidazole gave the cyclic thiocarbonate, which, upon reaction with triethyl phosphite, followed by deprotection, afforded 1. Treatment of 1 with phenyl methoxyalaninylphosphochloridate and N-methylimidazole gave 2. Catalytic hydrogenation of 1 gave 3. Hydrodediazoniation of 1 with tert butyl nitrite and tris(trimethylsilyl)silane gave 4. Compounds 1-4 were found to be inactive against the human immunodeficiency virus and exhibited minimal to no cytotoxic activity against the L1210 leukemia, CCRF-CEM lymphoblastic leukemia, and B16F10 melanoma in vitro. PMID- 15822620 TI - Co-transfection of messenger RNA and siRNA as a method to study the efficiency of siRNA. AB - The definition of an optimal siRNA results from the in vitro testing of several siRNA designed to specifically target a gene. Usually, such in vitro tests consist in the transfection of the several siRNA duplexes in a cell expressing stably the gene of interest. When a siRNA specific for a mRNA coding toxic proteins (certain transcription factors, transporters, toxins, cell cycle controlling proteins, etc.) must be tested, the generation of a target cell is difficult. Here we report a quick method to test the efficiency of a siRNA through its co-transfection with the targeted mRNA. This technique can be used as a fast method to test siRNA even when they target genes that cannot be stably expressed in the cells of interest. PMID- 15822621 TI - Concise synthesis and antiviral activity of novel unsaturated acyclic pyrimidine nucleosides. AB - Novel acyclic nucleoside analogues were designed and synthesized as open-chain analogues of neplanocin A. The coupling of the allylic bromide with pyrimidine bases using cesium carbonate afforded a series of novel acyclic nucleosides. The synthesized compounds 15-22 were evaluated for their antiviral activity against various viruses such as HIV, HSV-1, HSV-2, and HCMV. PMID- 15822622 TI - [Humor and illness]. PMID- 15822623 TI - [How a short life can be enriching!]. PMID- 15822624 TI - [Naturopathic therapy of respiratory tract diseases in childhood]. PMID- 15822625 TI - [What is behind burn-out syndrome?]. PMID- 15822626 TI - [Quality management in after-care--from theory to general practice]. PMID- 15822627 TI - [Good carbohydrates, bad carbohydrates? The glycemic index and its significance for healthy nutrition]. PMID- 15822628 TI - [Living on the "Children's Island", but completely in daily life]. PMID- 15822629 TI - [Society and delusions of beauty]. PMID- 15822630 TI - [Weaning from artificial ventilation]. PMID- 15822631 TI - Restless legs syndrome in 2004. AB - The restless legs syndrome (RLS) is a sensorimotor disorder characterised by an intense urge to move the legs and sometimes also other parts of the body, and accompanied by a marked sense of discomfort or pain in the affected body parts. This urge has a circadian pattern - it is most pronounced in the evening or during the night. RLS symptoms are relieved by movement. The pathophysiology of RLS is related to dopamine transmission insufficiency, low iron storage in substantia nigra neurons, and spinal cord dysfunction. RLS is idiopathic or secondary (usually associated with iron deficiency, end-stage renal failure, pregnancy and spinal lesions). One half of the patients with idiopathic RLS have positive family history of RLS. RLS is curable, though the choice of therapy and proper dosage titration may take a long time, and though the therapy may sometimes have to be changed owing to augmentation. The most important pharmacologic treatment used in RLS includes L-DOPA, dopamine agonists, opiates, anticonvulsants and benzodiazepines. Therapy improves significantly the condition in long-term at least in 80% of RLS patients. PMID- 15822632 TI - Axosomatic synapses on granule cells are preserved in human non-infiltrating tumour or lesion-related and mesial temporal sclerotic epilepsy, but markedly reduced in tumour-infiltrated dentate gyrus with or without epilepsy. AB - Granule cells of the human hippocampal dentate gyrus were examined. In controls, granule cells displayed somatic spines and cell nuclei with small infoldings. In addition, the cytoplasm of human granule cells always displayed lipofuscin. Subsurface cisterns of endoplasmic reticulum were frequently observed in the human granule cells. Two types of axosomatic synapses were found; most frequently symmetric and less frequently asymmetric. Many of the axosomatic synapses were isolated by glial processes in tumour or lesion-related epileptic patients, but the ultrastructural characteristics of granule cells were not different from those of the control patients. Large bundles of reactive astroglial fibres appeared regularly in all layers of the dentate gyrus. In tumour infiltrated hippocampi, glial processes dominated the neuropil and the number of perisomatic synapses was markedly reduced. Reduction in the number of perisomatic synapses did not correlate with severity and duration of seizures but did correlate with the malignancy of the tumour. It is suggested that reduction of perisomatic inhibition may not be a characteristic of granule cells in the epileptic human dentate gyrus. PMID- 15822633 TI - Changes of evoked epileptic seizures after the short term hypobaric hypoxia in the young rats. AB - Exposition to the acute intensive hypobaric hypoxia blocks the triggering mechanism of epileptic seizures only within a short time interval after the end of hypoxia period and it does not influence the progressive epileptogenesis in 12 day-old rats. In 25-day-old rats' hypobaric hypoxia suppresses the postictal depression elicited by the repeated stimulation in short interstimulus intervals and it increases the duration of epileptic seizures evoked by electrical stimulation of the sensorimotor cortex. PMID- 15822634 TI - Development of third molar in the Czech population. AB - The third molar development was studied on the basis of 1700 panoramic radiographs analysis of randomly selected individuals in the age ranging between 5-21 years inclusive. Each age group consisted of 100 probands--50 boys and 50 girls. The characteristics of the third molar development were following: time of dental follicle formation--dynamics of development--the most frequent stages of third molar development in individual age groups-- intraindividual differences and symmetry of development--incidence of 1-4 third molars agenesis--relation between the third molar development and the dental age--differences in the third molar development between individuals with all four third molars present and individuals with 1-3 third molars agenesis--ntersexual differences. On the basis of our analysis of seven stages of development, differences of this process between males and females and the changes in dynamics of development characterizing upper and lower jaw were described. The incidence of the third molar agenesis was statistically significant, being more frequent in boys (24.2%) than in girls (17.6%). The study showed, that when the third molar is not present at the age of thirteen, it could be taken in the Czech population as agenesis. Correlation between developmental stage of third molars and dental age was ascertained. With the exception of the agenesis, no other statistically significant intersexual differences were found. PMID- 15822635 TI - Two endocrinological anniversaries. AB - In 2004, the First Faculty of Medicine will commemorate two significant anniversaries related to the small, yet important field of endocrinology. On January 31, 2004 it has been twenty years since the death of the founder of the discipline in this country, professor Josef Charvat (6.8.1897 - 31.1.1984). The occasion was marked by the Dean of the First Faculty of Medicine, by the Prorector of Charles University and by the President of the Czech Academy of Science who jointly unveiled a memorial plaque dedicated to the founder-of Czech endocrinology placed on at the house in Ostrovni Street, where professor Charvat lived and died. During the brief ceremony, short quotes were read from the works of professor Charvat by the famous Czech actor Radovan Lukavsky and a speech was delivered by professor Vratislav Schreiber, the most notable of professor Charvat's students. On June 29, 2004, professor Schreiber himself, still full of vigour celebrated his 80th birthday. That day, too, is a significant anniversary in the history of Czech endocrinology. PMID- 15822636 TI - [Implementation of state policy in healthy nutrition of the population of the Russian Federation on the regional level: building up regional policy and regional programms. Methodical aspects of development and implementation. Part 1]. AB - Recommendations on development and implementation of regional policy and programs in the of healthy nutrition of the population are presented. PMID- 15822637 TI - [Studying the mechanism of antioxidant effect of vitamins and flavonoids]. AB - In work antiradical activity of vitamins and flafonoids in relation to hydroxyl radical is established. The most antiradical activity is established for flavonoids quercetin, routin, daidzein, biochanin A and luteolin-7glucosid, Among vitamins of group in the most antiradical activity have shown vitamins B12, B2, B5, Bc. In group of fat-soluble vitamins ascorbic palmitat, vitamin K1, vitamin A and beta-carotin have shown activity above, than a known antioxidant vitamin E. The carried out work allows to draw a conclusion that antiradical activity in the relation hydroxyl radical shows the broad audience of vitamins that probably is one of aspects of their therapeutic action. PMID- 15822638 TI - [The influence of vegetarian nutrition on the structure of mucosa in human colon]. AB - Structural and functional peculiarities of digestive tract's wall depending from the character of nutrition (vegetarian or mixed diets) are interpreted in morphology. Proper facts and results of investigations of other authors about morphological structure of tunica mucosa colonic according of type of nutrition are discussed. PMID- 15822639 TI - [Influence of the peptides, isolated from kumys and a sour milk of Mechikov on some parameters of immunity in animals, who underwent an immobilization-cold stress]. AB - The influence of peptide fractions, discharged of kumys (SK) and a sour milk of Mechnikov (PMS) on some parameters of immunity experimental animal, transferred a stress is investigated. The outcomes of examinations have shown, that SK promotes correction of immune violations called by action of a stress. So the introduction SK animal one day prior to a stress renders expressed immunoregulatory an operation on a content of neutrocytes and metabolism in them, depending from Oxygenium, promotes a raise of number of T lymphocytes in a blood animal transferred a stress. The peptide fraction, discharged of a sour milk of Mechnikov (PMS) does not render essential influence on the above-stated parameters. PMID- 15822640 TI - [Analysis of vitamin C in the biologically active supplements to food and food products enriched with micronutrients]. AB - Peculiarities in Vitamin C analysis of biologically active supplements to food and food products enriched with micronutrients are discussed. Visual titration method and spectrophotometric determination based on the utilization of sodium 2,6-dichlorophenolindophenolate are shown to be the most favourable oner. A special attention is paid to the nature of extragents used and protection against undesirable effect of some food components. PMID- 15822641 TI - [The nutrition analysis of the patients with cardio-vascular diseases]. AB - Authors of article have recently analyzed the actual food intake from patients with cardiovascular diseases with the help of a new computer program, based on frequency method. It was found a disbalance of intake main food components in different groups of patients. PMID- 15822642 TI - [Thiamin and ribovlavin status in populations of Arkhangelsk]. AB - It was been performed the epidemiological study of Thiamine and Riboflavin status of 3579 inhabitants in Arkhangelsk. Establish by 49.6% man and 47.4% woman have lower provision of thiamin. Lack of riboflavin reveal by 23.6% man and 21.7% woman. The analysis of the effect of seasonality on vitamins content shown the worst thiamin level in examined population in January-February and in September October. The worst Riboflavin content observed in examined population in December January and in July-August. PMID- 15822643 TI - [Moscow schoolchildren zinc status: correction of zinc insufficiency by organic zinc compounds]. AB - The paper presents the results of study of zinc hair content in 43 Moscow schoolchildren, aged 9-11 years of both sex (May 2003 to December 2003). Significant drop of zinc hair content was established in 67% of schoolchildren. It was shown that the organic form of zinc enchanted zinc hair content. The findings will be used to implement preventive and health-promoting measures among schoolchildren. PMID- 15822644 TI - [The influence of nutrition and environment on the nonspecific resistance of the organism and physical development of children]. AB - Decrease of adaptable opportunities at children from ecologically unsuccessful region with a defective diet is marked. Attributes deceleration, tendencies to leucopenia, lumphopenia, monocytopenia, reduction of the maintenance of hemoglobin are revealed. Change of structurally functional parameters of erithrocyto's membranes, infringement of a parity about and antioxidant processes is typical. Decrease of quality of health causes necessity of realization of improving actions. PMID- 15822645 TI - [Influence of infra-red and super high frequency heating on food value of the beef meat]. AB - In clause results of research of influence infrared and super high frequency heating on amino acid, fatty fabric and mineral; substances fresh beef are shown meat, after infra-red and the super high frequency of processing, also are shown influence of various modes infra-red heating of processing on amino acid of meat. Advantage of an infra-red way of processing is shown in comparison with super high frequency heating. PMID- 15822646 TI - [Effectiveness of using the biologically active additive to food from Laminaria in adolescents during complex treatment of the pulmonary tuberculosis]. AB - Main component of biologically active additive to food on the basis of sea alga Laminaria of which are copper derivatives of a chlorophyll. Application of this biologically active to food in complex with tuberculosis chemotherapy of 48 adolescents (14-17 years old) at a sanatorium stage of treatment was investigated. The comparison group were 30 patients, receiving chemotherapy without utilize of biologically active to food. It is established, that mentioned biologically active additive to food promotes favorable radiological dynamic of process, improvement of functional activity T-lymphocites and the contents of substances of low and average molecular weight and malonic dialdehyde in plasma. PMID- 15822647 TI - [The influence of the new beverage, fortified by vitamins and mineral substances on the mineral metabolism in children-invalids with locomotion disorders]. AB - The article accounts for the results of a study of influence of the drink on the mineral metabolism indices in children-invalids with locomotive disturbances. The clinical observation and biochemical study revealed the efficacy of use of this food supplement to achieve quick improvement of calcium metabolism in children with the above mention disease. PMID- 15822648 TI - Pregnant women's beliefs and influences regarding exercise during pregnancy. AB - OBJECTIVE: To sample women's exercise habits and beliefs regarding exercise during pregnancy and to determine patient awareness and physician communication of ACOG's 2002 guidelines for exercise during pregnancy. METHODS: A 28-item questionnaire was completed by 211 patients at a private obstetric and gynecology (Ob/Gyn) group practice during a 4-week study period. RESULTS: Ninety-five percent of the women believed that participating in prenatal exercise was helpful. There was a significant correlation (P<0.000) between a woman's beliefs regarding the benefits of prenatal exercise and whether or not she chose to exercise during pregnancy. Forty-one percent of women surveyed and 41.6% of women who exercised indicated that their doctor had the most influence on their beliefs regarding exercise. Approximately 92% of the women who discussed exercise with their physicians were encouraged by them to exercise. In addition, women who were encouraged to exercise by their physician were more likely (P=0.005) to exercise than those who were not. Only 9.8% of the 133 women who talked to their physician reported that their physician helped them develop an exercise program. Over 69% of women were advised by their physician to place limitations on their exercise habits according to ACOG's 1985 guidelines rather than the current guidelines. A significant relationship was found between BMI and whether or not women exercised (P=0.001). Despite the previously known correlation between BMI and gestational diabetes, we failed to find a significant relationship between BMI and whether or not their physician encouraged them to exercise (P=0.531). CONCLUSION: A majority of women studied believed that exercise will have a positive impact on their pregnancy, are deciding to exercise during their pregnancy, and indicated that their doctor had the most influence on their beliefs about exercise during pregnancy. Because women's behaviors are influenced by their physician's instructions, physicians should be prepared to counsel their healthy ante-partum patients on the benefits of prenatal exercise and be informed of ACOG's current guidelines. PMID- 15822649 TI - Refining the diagnosis: the true problem. PMID- 15822650 TI - Whitfield reflections. PMID- 15822651 TI - 150 years of caring--a brief history of Mississippi State Hospital. PMID- 15822652 TI - [The public volunteerism of Catherine Vautrin. Interview by Catherine Maisonneuve]. PMID- 15822653 TI - [Winter epidemics, lessons from the dog-days]. PMID- 15822655 TI - [Six keys to the "handicapped" law]. PMID- 15822656 TI - [Preventing accidental falls in the elderly]. PMID- 15822657 TI - [Promoting awareness of the seriousness of falls]. PMID- 15822658 TI - [Risk factors for falls]. PMID- 15822659 TI - [Screening and preventing falls in elderly persons at risk]. PMID- 15822660 TI - [Hip protector system to prevent upper femoral fractures]. PMID- 15822661 TI - [The faller at home]. PMID- 15822662 TI - [Making the first steps in the institution]. PMID- 15822663 TI - [Elderly persons, hold your heads high!]. PMID- 15822664 TI - [An equilibrium workshop--prevention of falls]. PMID- 15822665 TI - [Physically adapted activities and prevention: equilibrium workshops--fall prevention]. PMID- 15822666 TI - [When a fall is survived....]. PMID- 15822667 TI - [Osteoporosis always predicts other problems]. PMID- 15822668 TI - [Aging in beauty!]. PMID- 15822669 TI - [Hearing aides in the drawer]. PMID- 15822670 TI - [Benchmarking Alzheimer's dementia in institutions]. PMID- 15822671 TI - [Sense of care in geriatrics]. PMID- 15822672 TI - Shortfall, not windfall, greets PT/INR coverage. PMID- 15822673 TI - Seeing is believing as AP labs flip digital switch. PMID- 15822674 TI - [Coxiella burnetii infections as the public-health problem]. AB - THE AIM OF WORK: To evaluate the incidence of human Q-fever in individual regions of Bosnia and Herzegovina, to determine the prevalence and areas at risk of Q fever incidence, to assess Q-fever seroprevalence and to demonstrate seroprevalence ratings in regard to relevant criteria (sex, age, location of residence). METHODS: The research has involved 708 individuals from different cantons, whose sera were tested in Laboratory for Specific Diagnosis of Human Q fever on the Microbiology Desk of Medical Faculty of University in Sarajevo, during the period from July 2000-December 2003. Sera were tested by Indirect Immunofluorescence Assay (IFA), using commercial test kit, reagents for Q-fever IFA IgM and IFA IgG for in vitro diagnostic use (Focus technologies). THE RESULTS: Specific anti-C. burnetii antibodies were detected in 249 (35.2%) individuals, whereof 75 (30.12%) female individuals, and 174 (69.9%) male individuals. Individuals suffering from Q-fever were mostly included in 31-40 aged group (26.8%). Most seropositive individuals were from Zenica-Doboj Canton (35.4%), then from Herzegovinian-Neretvian Canton (22.9%), from Sarajevo Canton (14.5%), from Una-Sana Canton (11.6%), and from Middle-Bosnian Canton (11.6%). CONCLUSIONS: In our four-year study, it is serologically demonstrated that Q fever is imminent in Bosnia and Herzegovina and that it is going to be the actual concern of public health. The number and distribution of seropositive individuals suggest that Q-fever is endemic phenomenon in Bosnia and Herzegovina. PMID- 15822675 TI - [Laboratory-immunology indicators according as prognostic factors of patients with colon and rectum cancer]. AB - INTRODUCTION: The occurrence of colon and rectum cancers has risen gradually in the past twenty years. In order to make a precise forecast about a patient's life expectancy and/or treatment, the factors of prognosis and outlook are used in oncology. These can be classified as clinical-surgical, pathological-histological and laboratory-immunology. PURPOSE OF THE WORK: To establish the significance of the laboratory-immunology indicators, carcino-embryonic antigen (CEA), proliferate nuclear antigen (PCNA) as well as Ki67 and p53 in the survival rate of patients who have colon or rectum cancer. PATIENTS AND METHODS: In "Sveti Vracevi" Hospital in Bijeljina 33 patients suffering from cancer of the left half of the colon and rectum received treatment from 1st January 2000 to 1st January 2003. They all had immunohistochemical analysis of CEA, PCNA, Ki67 and p53. RESULTS: The patients with positive immunohistochemical CEA, PCNA and p53 values had statistically shorter life expectancy in comparison with the patients with negative CEA, PCNA and p53 values. There was not statistically significant difference in the survival rate of patients with positive immunohistochemical Ki67 values in relation to the patients with the negative values. CONCLUSION: Immunohistochemical analysis is not used in the routine analysis of colon and rectum cancers. Establishing them gives enough information on the patient's survival rate. PMID- 15822676 TI - [Rupture of the uterus as a complication in delivery after cesarean section]. AB - INTRODUCTION: In present clinical practice there is an increasing number of patients with delivery with previous cesarean section which is frequent reason for the repeated one. The most serious complication in labour with previous cesarean section is the rupture of the uterus which is as frequent in labour by natural way as in repeated cesarean section. This is a reason that many of obstetricians do not support attitude "Once cesarean section, always a cesarean". THE AIM: In a retrospective study in five years period beginning January 1, 1998, we analyzed total number of deliveries, total number of cesarean section, way of deliveries with previous cesarean section and frequency of complications. METHODS: We used Chi2 test and Contigency table. RESULTS: In analyzed period from January 1, 1998 through December 31, 2002, we completed 24,194 deliveries at Ob/Gyn Clinic in Tuzla. Out of this number we performed 4073 cesarean sections or 16.83%. Out of 24,194 deliveries we had 1328 (5.49% of all deliveries) deliveries after performed cesarean in previous pregnancy. Out of 1328 deliveries following cesarean we completed by natural way 323 pregnancies or 24.33% and by repeated cesarean 1005 or 75.67%. Previous one cesarean had 1191, previous two cesarean 126, and previous three cesarean had 11 women. Out of 1328 deliveries by previous cesarean we had 12 (0.90%) or 1:111 cases, complicated by intrapartum rupture of the uterus. It was significantly more than in group of deliveries by unscarred uterus (1:16,849 deliveries). Three of 12 ruptures of the uterus were in group by natural way of delivery and 9 cases in group by repeated cesarean, no statistical difference (p>0.05). Out of 12 cases complicated by rupture of the uterus, ten cases were incomplete and two complete rupture of the uterus. CONCLUSION: Repeated cesarean section by his complications imposes the careful choice of indication for primary cesarean section and by this way make maternal mortality and morbidity significantly lower, specially in the light of the fact that only 24.33% deliveries following cesarean is completed vaginally. The most frequent complication was rupture of the uterus in relation 1:111 deliveries. PMID- 15822677 TI - [Disability caused by essential hypertension complications in Sarajevo Canton in the period 1998-1999]. AB - The aim of this study was to assess working ability of persons suffering from some of essential hypertension (EH) complications in Canton Sarajevo in the period of two years (01.01.1998-31.12.1999). Recorded complications included: congestive heart failure (CHF), heart attack (HA), angina pectoris (AP) and cerebrovascular disease (CVD). The study included 606 persons with EH with clinically verified complications, out of whom there were 416 males of average age of 53.4 (+/- 14.7) years, and 190 females of average age of 50.3 (+/- 12.8) years. There were 377 (62.2%) persons assessed with complete loss of working capability, 126 (20.8%) with limited working capability and only 103 (16.9%) were found without disability in regard to the tasks required in their job description. It was confirmed that EH complications have significant impact to disability among insurants in their most productive working age. CHF is the leading cause of complete loss of working capability, particularly in the group of labour workers with low qualification structure, followed by CVD, HA and AP. Disability-loss and limited working capability was dependant on sex, age, type of complication, qualification structure and the type of occupation. The results of this study indicate urgent need for establishing a more efficient concept of primary prevention as well as better and comprehensive treatment of the EH in order to reduce morbidity and disability among the working age population. PMID- 15822678 TI - [Depression among students of the Medical Faculty and doctors in Mostar]. AB - To assess out how many students and doctors have a mild, a moderate and a gravely depressive episode. SUBJECTS: The research was carried out among the students and doctors of the Medical Faculty in Mostar, Bosnia and Herzegovina, in March 2003. Out of the total number of 255 medical students and 115 doctors of, 200 (78.4%) medical students and 90 (78.3%) doctors participated in the research. The were analysis in accordance with DSM-IV criteria. RESULTS: One hundred and twenty (41.4%) participants experienced mildly depressive episode, and 52 (17.9%) participants experienced moderately depressive episode. Medical students were more depressive (66.5%) than the doctors (43.3%, p=0.001). A majority of the mildly depressive, and in particular the moderately depressive respondents, reported about a mild and moderate, respectively, intensity of disorder of mood loss of interest for the usual activities, fatigue, and the loss of energy. CONCLUSION: Medical students and doctors constitute a vulnerable professional group, having a high percentage of depressive disorders. PMID- 15822679 TI - [Criteria how to choose adequate methodology and relevant variables for assessment of quality of primary healthcare]. AB - INTRODUCTION: Research of satisfaction of patients in the area of primary health care (PHC) and especially criteria how to choose best methodology and the relevant variables guided to the situation where activities were focused on description of the basic structure and processes of health care rather than on identification of needs for services and on result based assessment and satisfaction of patients with provided service. To give advantage to any of methodologies is inexplicable problem and depend of factors beyond of health service control. PURPOSE: Aim of this paper is to review existing measurements of healthcare for ones who intend to use measurements of quality of healthcare for clinical and research purposes and to recommend criteria how to choose adequate methodology and relevant variables for assessment of quality of primary healthcare. METHODOLOGY: For our study we used experience of the Cathedra for Medical Informatics, Medical faculty, University of Sarajevo with emphasis on research of quality of PHC. We selected over 3,000 users of PHC (patients-adults) in PHC units in ten cantons of Federation of Bosnia and Herzegovina. RESULTS: Statistically significant results we got when we crossed general attitude of patients about quality of healthcare with the following variables: place where patient receive healthcare, consultation with patients about patients' rights and life styles. We have not got statistically significant results when we crossed the following variables with general attitude of patients: distance from place of living to place where patients get healthcare, examination schedule in the units of PHC, continuity of healthcare, organization of field service and possibility to choose physician or nurse. CONCLUSION: Very often researchers create their own methods of measurement instead to carry out systematic review of existing ones to enable them to find appropriate one. In this paper we recommended guidelines for finding the most appropriate methods of health measurement using experience of the Cathedra for Medical Informatics, Medical faculty in Sarajevo. PMID- 15822681 TI - [Aromatase inhibitors--new horizons in breast cancer endocrine therapy]. AB - The knowledge that breast cancers in a great percent of patients have specific estrogen (ER) and progesterone (PgR) receptors, which made them hormonal dependent, induced the development of a large choice of hormonal drugs for the treatment. The mortality rate of hormonal responsible breast cancer has been drastically decreased by different sequential hormonal manipulations. It is mostly due to tamoxifen, which is an ER blocator. As the synthesis of estrogens in postmenopausal women occurs through the aromatization of androgens, the third generation of new nonsteroidal aromatase inhibitors (AI), have been showed as strong drugs for endocrine therapy. Since they showed the advantage over tamoxifen in advanced breast cancer of postmenopausal women, there are currently the investigations of their value in early breast cancer adjuvant therapy, as well as the side-effects of the therapy. So far, tamoxifen remains a gold standard of adjuvant hormonal therapy for endocrine-dependent breast cancer. PMID- 15822680 TI - [Review on the etiology and treatment of bacterial pneumonias]. AB - The bacterial pneumoniae are the significant factor of morbidity and mortality all over the world. So USA are at the sixth place per mortality cause. The bacteria are in 60-80% of cases the causes of pneumonia in the communities. Therefore this is the review on the ethiology of the verification and treatment of pneumonias in the 17-month period in the General hospital "Sarajevo". In the period of the 1st January 2002 till the 31st May 2003 year, it was examined 105 sputa on the antibiogram, and in 12 cases (11.4%) was found the positive bacterial finding. Most often were gram negative bacteria in 7 cases (58.3%), while were found the gram positive in 5 patients (41.7%). Therefore that the majority of the examinees belonged to the risk group, and that were the patients with over 50 years of age 11 (78.6%), with the chronic obstructive lung disease 5 (35.7%), with the malignant disease 1 (7.1%) and with the coinfection of the urinary tract. E. coli was found in 3 patient (25.0%), Kl. Pneumoniae in 2 (16.7%), Ps. aeruginosis in 1 (8.3%), Citrobacter freundi in 1 (8.3%), and in 2 cases (16.7%) were isolated. Staphylococcus intermedius and Streptococcus beta haemolyticus gr. A and Str. Pneumoniae in 1 case (8.3%). To the patients with the positive finding of sputum on antibiogram were most often administered empirically cefalosporia of the third generation because it was about the more severe patients, then the combination of ampicillin with gentamicin. All were dismissed as recovered or in better condition. Pneumonias is in the risk groups are possible to prevent to the five-years pneumococcus vaccine when is indicated or by influenza vaccine when simultaneously was prevented the viral, that is the possible bacterially superinfected pneumonia. PMID- 15822682 TI - Circulating levels of prolactin in breast cancer patients. AB - OBJECTIVES: In order to assess diagnostic value of prolactin (PRL) in breast cancer (BC), we examined its serum levels and frequencies of its increase in breast cancer patients (BCP), and compared them to those in two controls. We also determined circulating levels of PRL in localised and advanced BC and calculated sensitivity and specificity of PRL in BC. PATIENTS AND METHODS: The main experimental group consisted of 47 female patients with histologically confirmed diagnosis of BC. The obtained results were compared to those in two control groups: clinically healthy women, and female patients with other types and locations of cancer. Serum levels of PRL were measured by means of radioimmunoassay. Results were processed by means of t-test and two way analysis of variance. RESULTS: The serum levels of PRL before treatment, as well as the frequencies of its increase, were significantly higher in BCP in comparison to controls (p<0.01, 0.02). The average circulating levels of PRL in patients with advanced BC were significantly higher (p<0.0001) in comparison to patients with localised disease. Sensitivity for PRL in BC was 50%, and specificity was 100%. CONCLUSIONS: Increased levels of PRL can be detected in the majority of patients with advanced BC. PRL has high specificity for BC, especially for metastatic BC, which leads to its diagnostic and prognostic importance in this disease. PMID- 15822683 TI - [Performing liver resections with or without Pringle manuever]. AB - Liver resections, because of its complexity, present a big challenge for every surgeon. A malignant diseases, which are the most frequent indications for this type of operations, additionally complicate job of the surgeons. Because the flag immunological system, in this type of patients, with reduced reserves of the liver, can have unforeseen postoperative consequence. The bleeding during and after operations presents one of the most difficult problems with liver resections. Introduction of the Pringle maneuver, reduce the risks for this operations to the minimum. The aim of this work at first is to analyze and compare functional parameters of the liver with or without intermittent hepatic pedicle clamping (HPC), for the purpose of the establishing invasivity and justification of its application. A statistical pattern was 30 patients with performed liver resections using intermittent HPC and 30 patients without using intermittent HPC, during liver resections. Results showed that performing liver resections with Pringle manuever do not have negative repercussions on the liver functions with less postoperative complications and postoperative hospital duration. PMID- 15822684 TI - [Analysis of indications in late pregnancy interruption on a two-year material]. AB - Indications for a late pregnancy interruption on a two-year material (2002-2003) were analyzed. In the stated period, there were 101 interruptions of advanced pregnancy in total, out of which 52 occurred in 2002 and 49 in 2003. Out of the total number of interruptions, social indications were the reason in 54 cases, which amounted to 53.46% of the whole material, wherein they amounted to 53.84% in 2002, and in 2003 to 53.06% of indications for late interruption. Average gestational age of patients was X=15 Weeks in 2002, and X=14 Weeks of pregnancy in 2003. Average age of patients in the first year of research was X=29, and in the second year X=28 years of age. Pregnancy interruption was carried out by prostaglandin induction. Average induction-abortion interval was X=9.2 hours for the first group, and X=8.2 hours for the second one. The increased number of social indications for late interruptions in advanced pregnancies is a result of difficult economic conditions of living, and medical ignorance as well. PMID- 15822685 TI - [Characteristics of cesarean sections at the University Hospital Mostar]. AB - We performed a retrospective, descriptive study in order to analyze characteristics of the cesarean sections at the University Hospital Mostar in year 2002. We analyzed cesarean sections incidence, general characteristics of the parturients and neonates, major indications for the cesarean section and postoperative course. During year 2002, there were 247 cesarean sections out of total of the 1510 deliveries. Total of 255 neonates were born by this operative procedure and 16 of them were twins. We used the operational, labour, neonate and parturients records in order to collect data. The incidence of cesarean sections in University Hospital Mostar in year 2002 was 16.4%. There were 5.3% elective procedures. Most of the parturients were in age group 25-35. More than half of the parturients gave birth for the first time. The major diagnoses for cesarean section were previous cesarean section, fetal distress, breech presentation, cephalo-pelvine dysproportion and dystocia. Most of the pregnancies terminated by cesarean section (87.3%) were in term. The majority of neonates had weight > or = 2500 g (93.7%) and first minute Apgar score > or = 7 (96.4%). Parturients stayed for a week in the hospital and were given antibiotic prophylaxis postoperatively. Each second parturient suffered from pyrexia of unknown origin postoperatively. Comparing to literature data, we found high incidence of cesarean sections considering total number of deliveries and pregnancy pathology. We think that proper measures of decreasing this incidence should take place in University Hospital Mostar. PMID- 15822686 TI - [Prevalence of human papillomavirus infection in Slovenian women with repeated Pap II smears]. AB - In the detection of precancerous lesions the cervical Papanicolaou smear screening is used in Slovenia and worldwide. Management of patients with repeat abnormal smears (Pap II) represents a great and complex clinical and public health problem. Repeated cytologic examinations are routine procedure in many countries, also in Slovenia, although the sensitivity of Pap smear testing in the detection of cervical intraepithelial neoplasia (CIN) II and III is relatively low. In cases of abnormal squamous cells and mildly dyskaryotic cells the presence of infections with high-risk HPV genotypes is being increasingly used as a complementary method to Pap smear testing. In the study we enrolled 148 cervical samples of women who within two years had three subsequent Pap II smears (abnormal squamous cells or mildly dyskaryotic cells). The prevalence of HPV infections was determined using three molecular tests: Hybrid Capture 2 (hc2) test and two variants of polymerase chain reaction (PCR-PGMY11/PGMY09 and PCR CPI/CPIIG). HPV infection was detected in 25.7% of women. In women aged < or = 30 years a statistically significant higher prevalence of HPV infections was found (37.8%) than in women aged > 30 years (20.4%). Our findings show that repeat Pap smear as the method of follow-up and detection of precancerous lesions of the observed population do not provide relevant results due to low prevalence of HPV infections in Slovenia, which indirectly indicates low sensitivity and specificity of Pap smear testing. In the detection of HPV infections, molecular methods are thus sensitive screening tests to be used complementary to cytologic tests in women with abnormal squamous cells and mildly dyskaryotic cells. PMID- 15822687 TI - [The treatment of multi-drug resistant tuberculosis in the Clinic for Pulmonary Diseases "Podhrastovi" Sarajevo]. AB - One of the greatest problems in the treatment of the tuberculosis is the treatment of drug-resistant cases. In particular, the treatment of patients with multi-drug resistance (MDR) i.e. resistance to at least isonisid and rifampicin, is long and costly, and reqiures the use of drugs which frequently cause severe, often toxic, reactions. Multi-drug resistance causes a great concern among tuberculosis experts and public health officials. The development of multi-drug resistance in high levels is a sign that a control strategies are failing. MDR-TB may become an obstacle to effective treatment of tuberculosis. TB patients with MDR strains requires individualized approach and expensive treatment in specialised units. Treatment outcome for MDR tuberculosis (MDR-TB) is often poor with low cure rates and increased fatality rates, these patients may go on infecting others in the community because they remain infectious for a prolonged period of time. Since drug-resistance develops because of inadequate use of drugs, the qualitative treatment of new cases of tuberculosis is a key element of tuberculosis control because effective treatment quickly renders the patient non contagious and therefore stops the transmission of illness. In the Federation B&H the DOTS (Direct Observed Short Course of Therapy) recommended by WHO has been in use from 1994. Although we had some multi-drug resistant cases but in very small number. Using new regimen of treatment which is long lasting and very expensive we succeeded to cure them. In this article we show the results of the treatment of these patients in three-year period (2001-2003) in the Clinic for Pulmonary Diseases and tuberculosis "Podhrastovi" Sarajevo. PMID- 15822688 TI - [Contemporary methods of prevention and treatment diabetic kidney disease]. AB - The number of people with diabetes is increasing worldwide. Diabetes is the leading cause of end-stage renal disease. Diabetic kidney disease has been reported to occur in 25-40% of people with type 1 or type 2 diabetes. Recent studies have demonstrated that the onset and course of diabetic nephropathy can be significantly improved by intervention in early stage of the development of this complication. In this article we will review the current recommendations regarding the prevention and treatment of diabetic nephropathy. PMID- 15822689 TI - A simple method of DNA extraction in solving difficult criminal cases. AB - AIM: We examined combined use in house method and DNA IQ system (Promega) procedure for the extraction DNA from biological trace in order to help the solving some crime cases. METHOD: Simple and efficient method for extraction of the DNA from biological trace samples included the well washed trousers and the knife without any visual biological trails (from some one hard criminal case). In this method there were used: centrifuge, microwave oven, shaker, chelex, proteinase K, DNA IQ System (Promega). RESULTS: The extraction of DNA could be done within 2.5 hours or over the night. The DNA prepared on this way was good quality and could be used for STR analysis. CONCLUSIONS: This combined, simple, and safe method could be used for extraction DNA from samples containing a minute amount of the biological trace. PMID- 15822690 TI - [Neurofibromatosis type 2--case report]. AB - Young female patient, diagnosed and treated by ophthalmologist and neurosurgeon, are presented. Through this case the basic characteristics of NF 2 are stressed. The different clinical implications and similarities of NF I as a neurocutaneous disorders and NF 2 as a central form of disease are pointed out. PMID- 15822691 TI - [Why B&H does not have law on health data protection?]. AB - Protection of private data is secured by number of laws in EU and in some aspects they consider clinical or medical data, or they are more common and therefore reflect on general data protection. Considering the fact that, there are no laws in Bosnia and Herzegovina on Data protection, neither Health Data protection, it could be concluded that, based on Law on free access to information in Bosnia and Herzegovina, patients cannot control dissemination of private and confidential information about their health and health status in general. The authors suggest creation of legislative in this area in Bosnia and Herzegovina in accordance with directive 95/46/EC of European Parliament and Council of Europe, which come into force in EU in 1997. PMID- 15822692 TI - Aftermath: CNA launches an RN-to-RN effort in Sri Lanka. PMID- 15822693 TI - Staffing ratios in sight for Massachusetts. PMID- 15822694 TI - Neither social nor secure: privatization schemes defy common sense. PMID- 15822695 TI - Staffing ratios smashing success down under. PMID- 15822696 TI - 'Conan vs. the nurses'. RNs step up challenge to Gov. Schwarzenegger. PMID- 15822697 TI - Building a new national RN movement: NNOC responds to RN needs. PMID- 15822698 TI - Gaining respect and self-determination in Chicago. PMID- 15822699 TI - Breaking away. RNs thrive out of shadow of ANA. PMID- 15822700 TI - Why is this woman smiling? The hard sell of bad medicine. PMID- 15822701 TI - Probation year raises difficult questions. PMID- 15822703 TI - Wound care: what every nurse should know. PMID- 15822704 TI - Outside influence. PMID- 15822705 TI - Caring without prejudice. Interview by Rebecca Sandiford. PMID- 15822706 TI - The NPSA recommendations to promote correct-site surgery. AB - NPSA recommendations to promote correct-site surgery have been published. This article considers the need for such guidance and highlights its implications for nurses. PMID- 15822707 TI - Plantar fasciitis. PMID- 15822708 TI - Nursing considerations for people with Marfan syndrome. AB - People with Marfan syndrome present with a range of health problems that require specialised nursing, medical and surgical care. They face many challenges throughout their lives - some of which may be life-threatening - and require continuing support and education from a specialist team. Nurses play a pivotal role in the care of people with Marfan syndrome and their families, encompassing not only medical and surgical interventions, but aspects of lifestyle such as diet, exercise and hobbies, as well as family planning and psychological well being. PMID- 15822709 TI - Metoclopramide. PMID- 15822710 TI - Recruiting and training senior nurses using a rotational model. AB - In order to tackle some of the recruitment and retention issues in 'hard-to-fill' community posts such as district nursing, Croydon Primary Care Trust tested a community rotational model. This article addresses some of the issues raised by the participants in the first wave of recruits. New and innovative ways of recruiting and retaining staff are needed in the present employment climate in order to meet the changing needs of clients and service provision (Newman et al, 2002). Planning and developing a vision for health and social care requires a strategy that supports individual development as well as the rapid development of the skills, knowledge and experience required to function in today's demanding clinical.. environment (Forbes et al, 2001). PMID- 15822711 TI - Managing sleep disturbances in children with learning disabilities. AB - The view that children with special needs are more likely to experience sleep disturbances is well recognised. various physical and psychological factors associated with the child's condition, as well as the influence that the child's disability has on parents' attitude and management, combine to increase the likelihood of the child developing sleep disturbances. However, these factors should not act as a barrier to trying to find successful solutions. PMID- 15822712 TI - Improving care of patients who have nausea and vomiting. PMID- 15822713 TI - Discreet products for children and teenagers with continence problems. AB - The National Service Framework for Children, Young People and Maternity Services (Department of Health, 2004a) estimates that 'at least 500,000 children suffer from nocturnal enuresis, and a significant number with daytime wetting and faecal incontinence'. The NSF also acknowledges that 'incontinence is distressing for children and young people; it can lead to bullying at school and cause emotional and behavioural problems'. PMID- 15822714 TI - Commissioning continence services for children and young people. AB - The publication of Good Practice in Continence Services (Department of Health, 2000) was a major step forward in encouraging primary care trusts to focus their attention on the modernisation of continence services. Efforts to modernise included the development of integrated continence services (Rigby, 2001) while others focused on influencing service development (Thomas et al, 2004; RCN/Continence Foundation, 2004). Targets in the National Service Framework for Older People (DoH, 2001) - specifically Target Two on patient-centred care - have influenced the development of integrated continence services. PMID- 15822715 TI - Understanding urine testing. AB - Urine is a natural waste product of the body and urine testing is a simple and quick way of monitoring disease and identifying new illness. The results from a urine test can prompt a change in a patient's medical management. Urine testing is not a new procedure: as far back as the ancient Egyptians there are descriptions of polyuria. In 7th Century AD, Protosharis suggested that it was important to look at the colour of urine and by 1674 Thomas Willis, a professor at Oxford University, recorded that the urine of his patients with diabetes tasted sweet (Beer, 1996). PMID- 15822716 TI - Reducing the risk of catheter-related urinary tract infection. AB - It is estimated that 10-12 per cent of hospital patients and four per cent of patients in the community have a urinary catheter at any one time (Stamm, 1998). Urinary tract infections (UTIs) account for almost half of all health care associated infection (HAI), and a significant number of these infections are related to the insertion of urinary catheters (Pratt et al, 2001). Recent research has estimated that a patient who has a catheter for 20 days is almost certain to develop a catheter-related UTI and that the cost of treatment of UTI is estimated as being 1,327 pounds sterling per patient (Roadhouse and Wellsted, 2004). Plowman et al (1999) have estimated that a UTI can extend the length of a patient's stay in hospital by six days. PMID- 15822717 TI - Nurse prescribing, urinary tract infection and older men. AB - I recently managed the care of a male patient with acute and uncomplicated symptoms of a urine infection (Box 1). If my patient was female I could have prescribed antibiotics, but unfortunately independent nurse prescribers cannot prescribe antibiotics to men with urinary tract infections. My patient had to wait for a doctor to visit who asked my advice and prescribed what I recommended. PMID- 15822718 TI - Working as a medic on an offshore oil rig. Interview by Clare Lomas. PMID- 15822719 TI - Is it wise to leave science to big business? PMID- 15822720 TI - Skimming the cream. PMID- 15822721 TI - Moratorium. PMID- 15822722 TI - Oklahoma tobacco policy-making. AB - OBJECTIVES: To document and analyze comprehensive Oklahoma state tobacco policy making trends and their link to public health trends. METHODS: A historical qualitative and archival content overview and analysis from 1985 to the present of previously secret tobacco industry documents. RESULTS: The 2002 Oklahoma State Plan for Tobacco Use Prevention and Cessation indicated tobacco use was the largest cause of annual preventable death in Oklahoma. The Oklahoma State Plan recommended a variety of proposals, including sales restrictions, youth access enforcement, repealing state preemption of stricter local tobacco control laws, adopting clean indoor air laws, increasing tobacco taxes, cessation programs, and anti-tobacco education efforts. CONCLUSIONS: Since 1985, the powerful tobacco lobby in Oklahoma has been highly successful in maintaining lower tobacco taxes and reduced regulations, which is a major impediment to the anti-tobacco public health goals of the Oklahoma State Plan and enhanced public health. PMID- 15822723 TI - Why should your practice perform compliance audits based on the latest OIG Work Plan? AB - Why have compliance audits, or medical record reviews, become an essential part of many progressive medical facilities? Because medical record reviews are known to improve reimbursement, documentation, and internally identify compliance issues that should to be promptly addressed, in order to protect the organization from substantial monetary penalties and legal sanctions by the federal government. This article discusses the importance of auditing based on the areas of focus recently identified in the OIG Work Plan. These include VA physician billing, pathology/clinical laboratory services, identifying referring physicians in Oklahoma that are sanctioned, high level E/M services, consultations, and red flags that are raised due to overuse of modifier -25 and -59. PMID- 15822724 TI - Memorandum to OSMA Board of Trustees and House of Delegates: unionization and collective bargaining study. PMID- 15822725 TI - Smoke-free beginnings: changing practice behaviors to improve the delivery of smoking cessation to prenatal patients in Oklahoma. PMID- 15822726 TI - Patient page #58. Tips for healthy aging. PMID- 15822727 TI - Specialty hospitals: the truth regarding their impact on healthcare. PMID- 15822728 TI - Delirium in the older hospitalized patient. PMID- 15822729 TI - [Angioplasty in the elderly]. AB - Due to increasing age in the general population, patients > 75 years are more and more often submitted to cardiac catheterization. These patients have, in general, more severe and diffuse coronary disease, more severe comorbidities, and a higher risk for periprocedural complications. Elderly patients have traditionally been excluded from most clinical trials of coronary interventions, and most often receive medical undertreatment in clinical practice. The basis of evidence for an early invasive strategy, as compared to optimal medical management, is therefore limited in these patients and the risk/benefit ratio is poorly known, both in the setting of acute coronary syndromes and of more stable coronary heart disease. A broad review of the literature is summarized in this paper, to help make therapeutic decisions in these patients. PMID- 15822730 TI - [Yentl syndrome. The underestimate of cardiovascular risk in women]. AB - Cardiovascular diseases are the leading cause of death for women in all countries of the world. Aging of the population and the high prevalence of risk factors among young and middle-aged women allow to hypothesize that this situation will continue in the future. Differences exist between women and men in the impact of risk factors, symptoms and therapeutic response. However, the main problem limiting prevention and control of cardiovascular diseases among women is gender inequality in health care. Myocardial infarction and stroke continue to be seen as "male" diseases and this view has deeply limited research and clinical management improvements. Furthermore, factors related to the socioeconomic environment strongly influence the development of cardiovascular diseases. For women, the ability to stop smoking, have a healthy eating and regular physical activity and live in a supportive psychosocial environment is strongly influenced by their level of income, education, role, control over their lives, culture, religion, access to health care. For the majority of women these factors represent the main barriers to cardiovascular disease prevention. The promotion of cardiovascular health among women could be accomplished only by removing all the obstacles to women's active participation in public and private life allowing them to share a full and equal role in economic, social, cultural and political decision-making processes. This strategy will not only improve women's health, but also that of children and men. PMID- 15822731 TI - [Cardiopulmonary exercise testing: the exercise stress test of the future?]. AB - Scientific evidence is mounting that cardiopulmonary exercise testing (CPET) offers different potential clinical applications in cardiology. In comparison with traditional ECG stress testing, CPET provides more accurate diagnostic information and prognostic insights as well, in particular in patients with chronic heart failure and primary pulmonary hypertension. The barriers that actually limit the diffusion of CPET among cardiologists are primarily "cultural", because modern ergospirometric tools are easy to use, and the time required for a CPET is not dissimilar from that of a standard ECG stress test. At present, time seems to be ripe for considering CPET as the new exercise stress test in cardiology. PMID- 15822732 TI - [Statistical notes. Hazard curves of sudden death: their role in the cardiologist's clinical decision]. AB - In long-term studies researchers are mainly concerned with occurrence of death during the follow-up period. This statistical note is focused on survival analysis which is the main tool to process this kind of data. Survival curve, cumulative mortality curve and hazard curve are here introduced together with an appropriate effect indicator: hazard ratio. In particular, the use of the latter is shown by resorting to the randomized controlled clinical trial TARGET. PMID- 15822733 TI - [Does reluctance to perform mouth-to-mouth ventilation exist among emergency healthcare providers as first responders?]. AB - BACKGROUND: Only 30% of survivors from out-of-hospital cardiac arrest receive basic life support (BLS) before the arrival of emergency personnel. This is also due to reluctance to perform BLS, especially mouth-to-mouth ventilation without barrier devices in victims who are unknown to the rescuer (either layperson or healthcare provider). METHODS: To evaluate the incidence of reluctance to perform mouth-to-mouth ventilation without barrier devices and its consequences in a simulated BLS scenario proposed by a questionnaire to healthcare providers of critical area in a public general hospital. RESULTS: Answers were collected from 128 of 165 (77.5%) interviewed healthcare providers. Physicians were 46 of 128 (35.9%), professional nurses were 78 of 128 (60.9%) and 4 of 128 (3.2%) were other health workers devoted to patient assistance. Seventy-five of 128 (58.6 %) were reluctant to perform mouth-to-mouth ventilation without barrier devices; 68 of 75 (90.6%) would perform BLS only by chest compression. Compared with non reluctant providers, they would have been available to perform assisted ventilation by non-validated alternative methods (54.2 vs 18.8% respectively, p < 0.001). Seven of 75 (9.6%, no physician among them) would perform no BLS at all. The most significant predictors of reluctance were age < 40 years (p = 0.07) and previous attendance of BLS-BLSD courses (p = 0.07). CONCLUSIONS: Reluctance to perform mouth-to-mouth ventilation without barrier devices is frequent and may reduce the number of potential BLS providers. Because of the concern about disease transmission between victim and rescuer, rescuers with a duty to respond such as healthcare providers should follow precautions including the use of barrier device also outside their workplace. When barrier devices are unavailable first responders should consider chest compression alone instead of not performing any BLS maneuvers. BLS training should help give a greater emphasis on this topics. PMID- 15822734 TI - [Stressful life events and depressive disorders as risk factors for acute coronary heart disease]. AB - BACKGROUND: Several studies outlined the role of stressful life events in the pathogenesis of coronary heart disease. It has recently been emphasized the role of depression, both clinical and subclinical, in the course of myocardial infarction. The relationship between recent life events, major depression, depressive symptomatology and onset of acute coronary heart disease has been less considered. METHODS: Ninety-seven consecutive patients with first episode of coronary heart disease and 97 healthy subjects matched for sociodemographic variables were included. All patients were interviewed by Paykel's interview for recent life events, a semistructured interview for determining the psychiatric diagnosis of mood disorders, a semistructured interview for demoralization. Patients were assessed while on remission from the acute phase. The time period considered was the year preceding the first episode of coronary heart disease, and the year before interview for controls. RESULTS: Patients with acute coronary heart disease reported significantly more life events than control subjects (p < 0.001). All categories of events (except entrance events) were significantly more frequent. Thirty percent of patients were identified as suffering from a major depressive disorder; 9% of patients were suffering from minor depression, and 20% from demoralization. Even though there was an overlap between major depression and demoralization (12%), 17% of patients with major depression were not classified as demoralized and 7 % of patients with demoralization did not satisfy the criteria for major depression. Independently of mood disorders, patients have a higher (p < 0.001) mean number of life events than controls. With regard to life events, the same significant difference (p < 0.001) compared to controls applied to patients with and without mood disorders. CONCLUSIONS: Our findings emphasize the relationship between life events and acute coronary heart disease. These data, together with those regarding traditional cardiac risk factors, may have clinical and prognostic implications to be verified in longitudinal studies. PMID- 15822735 TI - [The opinion of Roberto Antonicelli entitled "Hospital cardiology between specialty and outpatient diagnostics: a proposal for solution"]. PMID- 15822736 TI - [In the 2004 September issue of the Italian Heart Journal Supplement appeared an article of Del Colle et al. entitled "Assessment of cardiovascular risk in hypertensive patients: comparison among scores"]. PMID- 15822737 TI - [Short- and long-term effects of internal carotid arteries stenting in high-risk patients]. AB - The efficacy of endovascular treatment of internal carotid arteries (ICA) stenosis (20 patients) has been compared to that of conventional carotid endarterectomy (25 control cases). ICA stenting was performed in case of stenosis, narrowing arteries to > or = 60% in "symptomatic" patients and to > or = 80% in "asymptomatic" ones. Angiographic indices revealed favorable outcome after stenting in 100%. Mean extent of residual stenosis was 8.3 +/- 0.8%. Significant complications (stroke/fatal outcome) after 22 procedures developed in 3 (13.6%) cases. Mortality was estimated as 4.5%. Taking stroke and fatal cases into account, the interventions were successful in 86.4% cases. The duration of follow-up study was from 1 to 48 months, with 1 case resulting in death and 2--in development of acute disturbance of cerebral blood circulation of contralateral ICA. Short- and long-term results of the intervention did not significantly differ between the study and control groups thus indicating ICA stenting as an alternative to carotid endarterectomy procedure. PMID- 15822738 TI - [Pharmacological therapy of sialorrhea in patients with motor neuron disease]. AB - A comparative trial of amithryptiline and dysport (botulinic toxin type A) in the treatment of sialorrhea in patients with motor neuron disease (MND) was conducted in 10 MND patients with sialorrhea, of whom 5 were treated with subcutaneous injections of Dysport and 5 with Amithriptiline, and 6 controls without salivary dysfunction. Gravimetry and scintigraphy of salivary glands were used before and after treatment. Compared to controls, saliva production was significantly decreased in MND patients. Both amithryptilin and dysport used in mean therapeutic doses decreased sialorrhea with similar effect. However, 3 patients, receiving amythryptiline in dosage 50 mg/day, experienced side effects (constipation, accommodation disturbances, dry mouth, sleepiness and poor concentration). Reducing of amithryptiline dose, along with prescribing dysport, removed the side-effects in these patients, while sialorrhea did not increase. The authors concluded that due to high efficacy and low cost of amithryptiline therapy of sialorrhea proved to be a golden standard of palliative care in MDN. However, in these terms dysport can not be an alternative to amithryptiline in sialorrhea therapy. Nevertheless, in cases when amithryptiline treatment is accomplished with side-effects, the drug dosage can be reduced and combined with dysport. PMID- 15822739 TI - [A state of hemorheological system in patients subjected to carotid endarterectomy with atherosclerotic lesion of the major cerebral arteries]. AB - To study the influence of changes in the blood rheological indices and blood coagulation on development, progression and clinical course of restenosis and thrombosis, 62 patients (mean age 63.4 +/- 6.0 years), were followed up for 2-15 years after carotid endarterectomy. Above 50% (maximum 70%) restenosis was observed in 7% cases. No critical postoperative restinosis was observed. In 12% patients, restenosis was accompanied with carotid artery occlusion and hematocrit, with fibrinogen (FG) being significantly increased in the absence of fibrinolytic activity (FL) elevation, indicating hemorheological disorders and homeostasis activation in such patients. Long-term follow-up showed that viscosity blood characteristics were elevated but functional activity of platelets and FA in patients did not differ from those in healthy subjects. At the same time, FG was high and fibrinolysis index was lower than mean statistical value that indicated predominance of blood procoagulation activity. PMID- 15822740 TI - [Peculiarities of attention in families with schizophrenia: correlation between psychological and neurophysiological characteristics]. AB - A relationship between neurophysiological and psychological characteristics of attention was studied in 18 patients with schizophrenia and their 34 mentally healthy relatives: parents and siblings (20 subjects) and children (14 subjects). In patients, a decrease of P300 auditory evoked potentials significantly correlated with disturbances of attention stability and volume as well as characteristics of involuntary attention. At the same time, in the groups of relatives the anomalies of attention stability and attention in conditions of prolonged concentration were positively related to P300 latency. PMID- 15822741 TI - [Structure of peripheral blood platelets surface in patients with amyotrophic lateral sclerosis and multiple sclerosis]. AB - Using scanning electronic microscopy of peripheral blood platelets in 32 patients with amyotrophic lateral sclerosis (ALS) and 6 patients with multiple sclerosis (MS), platelets activation, emerging in pseudopodiums formation, aggregation and lysis, was found. Platelets activation was more pronounced in ALS than in MS. It was most markedly seen for quantitative evaluation of the above traits. The features of platelet activation found in the study support the evidence for considering ALS as a systemic disorder being characterized by a broad spectrum of alterations of biological processes. PMID- 15822742 TI - [Markers of endothelial dysfunction in attack-like schizophrenia]. AB - The activity of leukocyte elastase (LE) and two markers for vascular endothelial cells (EC) dysfunction/lesion--von Willebrand factor (WF) and C-reactive protein (C-RP)--was determined in the blood plasma of 22 male patients, aged 23.7 +/- 5.5 years with attack-like schizophrenia (ALS) and 24 matched controls. A significant increase of WF, C-RP and LE was found in the patients compared to controls indicating EC dysfunction and/or lesion in ALS schizophrenia. Positive correlations between LE activity and WF concentrations as well as between WF and severity of clinical symptoms in ALS patients imply a vascular layer lesion of the blood-brain barrier. PMID- 15822743 TI - [An absence]. PMID- 15822744 TI - [A case of pseudotumorous course of brain demyelinating disease]. PMID- 15822745 TI - [Polyarteritis nodosa and cerebrovascular disorders]. PMID- 15822746 TI - [Betaserc and improvement of life quality in war veterans]. PMID- 15822747 TI - [Peculiarities of mineral metabolism in patients with epilepsy]. PMID- 15822748 TI - [Stenting of internal carotid arteries]. PMID- 15822749 TI - [Microsatellite DNA]. PMID- 15822750 TI - [Novel bacterial DNA polymerases--their role in genomic damage tolerance]. PMID- 15822751 TI - [Tight binding transition state analogues of purine nucleoside phosphorylase- meaning, design and properties]. PMID- 15822752 TI - [Selected aspects of biogenesis of respiratory chain proteins in mammalian mitochondria]. PMID- 15822753 TI - [Structure and function of dynamin and dynamin-like proteins]. PMID- 15822754 TI - [Fibrinolytic system in atherogenesis]. PMID- 15822755 TI - [Secretory function of adipose tissue]. PMID- 15822756 TI - [Role of endothelial caveolae in the endocytosis of advanced glycation endproducts]. PMID- 15822757 TI - [Green fluorescent protein (GFP)--structure, features and applications in molecular biology of plants]. PMID- 15822758 TI - Position paper: Single-dose activated charcoal. AB - Single-dose activated charcoal therapy involves the oral administration or instillation by nasogastric tube of an aqueous preparation of activated charcoal after the ingestion of a poison. Volunteer studies demonstrate that the effectiveness of activated charcoal decreases with time. Data using at least 50 g of activated charcoal, showed a mean reduction in absorption of 47.3%, 40.07%, 16.5% and 21.13%, when activated charcoal was administered at 30 minutes, 60 minutes, 120 minutes and 180 minutes, respectively, after dosing. There are no satisfactorily designed clinical studies assessing benefit from single-dose activated charcoal to guide the use of this therapy. Single-dose activated charcoal should not be administered routinely in the management of poisoned patients. Based on volunteer studies, the administration of activated charcoal may be considered if a patient has ingested a potentially toxic amount of a poison (which is known to be adsorbed to charcoal) up to one hour previously. Although volunteer studies demonstrate that the reduction of drug absorption decreases to values of questionable clinical importance when charcoal is administered at times greater than one hour, the potential for benefit after one hour cannot be excluded. There is no evidence that the administration of activated charcoal improves clinical outcome. Unless a patient has an intact or protected airway, the administration of charcoal is contraindicated. A review of the literature since the preparation of the 1997 Single-dose Activated Charcoal Position Statement revealed no new evidence that would require a revision of the conclusions of the Statement. PMID- 15822759 TI - MS 04-044: demographic features of drug and chemical poisoning in northern Malaysia. AB - Acute poisoning is a significant health problem all over the world. In Malaysia, nationwide data on poisoning pattern is scarce and incomplete. The objectives of our study were to determine the pattern of acute drug and chemical poisoning at Penang General Hospital (PGH), in the northern region of Malaysia, and to compare poisoning characteristics between different ethnic groups. The study was a retrospective case review of all poisoned patients admitted to PGH during the years 2000-2002. We collected data concerning demographic parameters of patients, information about the agent(s) implicated, and circumstances surrounding the event. There were 493 poisoning incidents. Nearly two-thirds of the poisoned cases involved female patients. The predominant mode of poisoning was intentional (51.5%). The age group 15.1-30 years ranked at the top, constituting 55.2% of all cases. Drugs were the predominant agents implicated. Among cases associated with drugs, paracetamol was the main causative agent (44.7%). Chinese patients constituted 37.7% of all poisoning cases, followed by the Indians (31.6%) and Malays (26.6%). Between ethnic groups, Indian patients were found to have the highest rate of poisoning admission of 75.2 per 100,000 persons. PMID- 15822760 TI - How tightly can a drug be bound to a protein and still be removable by charcoal hemoperfusion in overdose cases? AB - BACKGROUND: Charcoal hemoperfusion is a very effective extracorporeal treatment in cases of drug overdose. The volume of distribution of a drug is the most important factor in limiting the efficacy of the treatment. Typically, drugs with a small volume of distribution are more efficiently removed by this treatment. In contrast, the effect of the plasma protein binding properties of drugs on their removal is not well understood, but it is clear that this binding percentage is an additional factor that affects the treatment. OBJECTIVE: To clarify the role of protein binding of a drug on the effectiveness of charcoal hemoperfusion and to define a guideline for this treatment based on the percentage of drug binding. STUDY DESIGN AND METHODS: Twelve patients with drug overdoses involving 20 different (a total of 32) drugs were hemoperfused using a charcoal column at Japanese Red Cross Kumamoto Hospital in 2000. Immediately after the beginning of the treatment, the plasma concentrations of the drugs in the blood entering (A) and leaving (V) the charcoal column were determined. The extraction efficiency, (A-V)/A, of each drug was then calculated. RESULTS: The efficacy of drug removal through adsorption to activated charcoal was found to be dependent on the binding affinity which is related to the protein binding percentage. The relationship between the extraction efficiencies of a charcoal column and the plasma protein binding percentages of the drug(s) showed that drugs that were bound at levels of 90-95%, or less, were effectively removed from the blood. CONCLUSIONS: Charcoal hemoperfusion can effectively remove drugs with a protein binding percentage as high as 95%. In addition to the volume of distribution, the plasma protein binding percentage of the drug can be used as a determinant for clearance by hemoperfusion especially in cases of a drug that binds tightly to a protein. PMID- 15822761 TI - Mercury exposure of mothers and newborns in Surinam: a pilot study. AB - AIM: To study mercury levels in hair from mothers and newborns in Surinam. METHODS: Hair and urine was collected from thirty-nine mothers of different ethnicity and hair from their newborns. The women delivered in a large hospital in the capital. Mercury analysis was performed. RESULTS: Fourteen (36%) of the mothers had elevated Hg concentrations in the hair as compared with a reference US population. Thirty-one newborns (80%) presented with a higher mercury level in hair than their mothers. A positive correlation existed between the hair levels of the mothers and their newborns. Urine Hg concentrations were not elevated. CONCLUSION: Mercury accumulates during pregnancy in the unborn. Further study is needed to elucidate the background and consequences of this finding. PMID- 15822762 TI - Acute poisoning in adults in the years 1996-2001 treated in the Uludag University Hospital, Marmara Region, Turkey. AB - BACKGROUND: Acute poisonings are frequent causes of admission to emergency departments and these cases may have hazardous outcomes. METHODS: In the present study, medical records of 1818 poisoned patients admitted to Uludag University Medical School's Emergency Department between January 1996 and December 2001 were investigated. The age, sex, outcomes of the patients, and type of poisoning are described. RESULTS: . The mean age for females (63% of the patients) was 27 years, whilst the mean age of male patients was 31 years. The major types of poisonings were ingestions of medications (59.6%), mushrooms (3.3%), corrosives (2.5%), organophosphates (3.2%), and methyl alcohol (0.4%). Carbon monoxide accounted for 6.9% of intoxications. Approximately 65% of the patients survived, while the methyl alcohol and corrosive ingestions led to the highest fatality averages (100% and 14.8%, respectively). CONCLUSIONS: The demographic and diagnostic features of acute poisoning cases treated in our hospital are similar to those reported in the literature. Adults and women are in a high-risk group for acute poisonings and medicine poisoning, which is the most common type of poisoning. PMID- 15822763 TI - Hexavalent chromium ingestion: biological markers of nephrotoxicity and genotoxicity. PMID- 15822764 TI - Intravenous amitraz poisoning. AB - Amitraz, a derivative of dimethylformamidine, is an acariside and insecticide used to control parasites in animals. Amitraz inhibits monoamine oxidase and prostaglandin synthesis and is an alpha-2 adrenergic agonist. Xylene, a mixture of o-, m-, and p-dimethylbenzene, is widely used in industry. A 22-year-old woman was poisoned by an intravenous injection of 5-6 mL of an amitraz formulation (amitraz 12.5% + xylene 57.5%). Clinical findings were coma (Glasgow coma score 3), respiratory depression, hypotension, bradycardia, hematuria, and edema and hyperemia at the injection site. Although her coma and other symptoms quickly resolved, as has been seen in oral and dermal amitraz poisoning, intoxication with higher doses occurring from intravenous injection may result in more serious problems. PMID- 15822765 TI - Acute intoxication due to ingestion of vegetables contaminated with aldicarb. AB - Three members of the same family ingested vegetables treated with aldicarb. All three developed signs and symptoms of acetylcholinesterase inhibition and all recovered a few hours after the ingestion. Reports of toxicity from the ingestion of aldicarb-contaminated food are uncommon. Aldicarb is a potent pesticide which can only be used safely if governmental and industry regulations are followed carefully. PMID- 15822766 TI - Sea urchin envenomation. PMID- 15822767 TI - The use of toxicogenomic data in risk assessment: a regulatory perspective. AB - Recent advances in genomic research have provided new insights into mechanisms of toxicity evoked by drugs or xenobiotics. Several governmental and commercial organizations are at present actively building up databases containing large amounts of toxicogenomics information. With promises to improve our ability to characterize hazard, scientists are facing the challenge as to how to link patterns of gene expression and gene clustering to specific adverse effects of toxicants or classes of toxicants. The future of toxicogenomics lies in the robustness of the databases generated. Regulators are working together with institutes and industry in exploring the potential use of these databases in regulatory purposes. It is, however, anticipated that regulatory use of the toxicogenomic databases as supportive information in the assessment procedure of new drug applications will be on a case-by-case basis until the predictive value of the databases is firmly established. PMID- 15822768 TI - Hornet sting-induced toxic hepatitis. PMID- 15822769 TI - The revised position papers on gastric decontamination. PMID- 15822770 TI - [Conception of psychophysiologic support of military physician training]. PMID- 15822771 TI - [Characteristics of work stimulation of the medical personnel in the treatment and-prophylactic institutions]. AB - Social and economic relations of the last years that were formed in our country have lead to the changes in approaches to administrative activity. Under conditions of administrative-and-command guidance the patriotism and requirements of production discipline ensured the highly effective work. Today the economic relations between employer and personnel are of the first priority. These principles can be attributed to the military medical collective activity. The algorithm developed during the investigation for stimulating the employees working in the in-patient military medical institutions suggests the following performance of measure complex directed to the improvement of professional training of managerial staff working in the military medical institutions work with subordinates, fulfilment of motivation activity, summing up the medical staff work. PMID- 15822772 TI - [Preparation "Santa-Rus" as a component of the lactobacterium therapy of digestive tract diseases]. PMID- 15822773 TI - [Medical care rendered to the wounded with abdominal nonpenetrating injury during pre-hospital stage]. AB - Of 4477 lethal outcomes the death caused by ACI constituted 6.2%. At pre hospital, stage the lethality was 45.7%, including 67%--from massive hemorrhage, 19%--from shock, 14%--from reflex cardiac arrest caused by abdominal stroke. Of 157 ACI cases 70% were admitted at the peak of development of posttraumatic complications, 20%--in 24 hours after trauma. According to "VPH-SP" scale the severity of casualties' state was the following: 18% (up to 12 points), 43% (16 17 points), 31.2% (21-23 points), 12% (34-37 points), 5% (45-53 points). At admission 36% had a shock, 50.3%--continuous hemorrhage, 18%--peritonitis symptoms. According to the investigation results the main factors that have an influence on the outcome during ACI are the qualitative skilled medical care at the place of trauma with the use of analgetics and infusion therapy as well as qualitative surgical care. Type of transport and speed of the wounded delivery are of minor importance. According to the state severity during ACI 3 groups were selected: up to 10 points; 10-20 points; 21-45 points. For each group the necessary optimal volume of medical care at prehospital stage was recommended (the use of analgetics, infusion and hemostatic agents). To improve the quality of medical care rendered to ACI casualties the medical service adequate to the trauma is required. Besides the emergency care units should be provided with sanitary transport equipped with special apparatus and have the trained medical staff. PMID- 15822774 TI - [The modern principles of the complex therapy of acute cholangitis]. PMID- 15822775 TI - [The local recurrence of renal-cell carcinoma after nephron-sparing surgery]. AB - The possibility of local recurrence in the remnant of kidney is the main limitation of nephron-sparing surgery in the patients with renal-cell carcinoma. The results of conservative surgery were analyzed in 176 patients. The local recurrences were revealed in 6.3% patients (5.1% in the imperative group and 1.2% -in elective group). Cancer multifocality was the source of recurrence in 8 patients, positive surgical border of the tumor--in 3. The type of surgical intervention (enucleation, enucleoresection, resection) doesn't affect the frequency of tumor local recurrence. The careful monitoring with the help of modern methods of visual diagnosis during the patient whole life is the main problem in the prophylaxis of such pathology. PMID- 15822776 TI - [The efficiency of magneto-laser therapy used in excretory-and-inflammatory form of male infertility]. PMID- 15822777 TI - [The analysis of HIV-infection incidence among the persons of draft age in Ulyanovsk region]. PMID- 15822778 TI - [Organizational-and-law aspects of counteraction to HIV/AIDS spread among the servicemen]. PMID- 15822779 TI - [Organizational-and-law problems of rendering care to the wounded during the Great Patriotic War]. PMID- 15822780 TI - [The Novocherkassk military hospital is 85 years]. PMID- 15822781 TI - [The history of medical-and-prophylactic work and medical-and-prophylactic sciences in the Armed Forces of Russia]. PMID- 15822783 TI - [Pathogenetic features of respiratory distress syndrome in newborns with extremely low body weight]. AB - Congenital atelectasis and ascending infection around the fetuses were of importance in the pathogenesis of respiratory disturbances in 38 newborns with extremely low body weight. PMID- 15822784 TI - [Immunopathomorphology of the human placental barrier in the first trimester of pregnancy complicated by the inflammation of the birth-ways]. AB - The effect of ascending infection of birth-ways on transport of maternal immunoglobulins (Igs) through the placental barrier in humans during the first trimester of pregnancy was studied. The transport of Igs is seen already in 3.5 to 5-week-old embryos, and different cellular and biochemical compounds participate at each stage of this process. Transport of Igs through the trophoblast is carried out due to the secretory component (SC) and, perhaps, to some other receptors. Monocytes together with Igs penetrate into capillaries between the endothelial cells and are transported with the blood all over the body. It seems that SC and other receptors help Igs to penetrate into capillaries through the endothelium. Further, Igs are transported with erythroblasts. In the development without infection the transport of IgG was seen in all cases studied. Inflammation of the birth-ways is accompanied by an increase in transport of all Igs, already in early embryogenesis. Three groups were distinguished: 1) abortions without inflammation; 2) cases with signs of moderate inflammation (endometritis, deciduitis); 3) cases with intensive inflammation with necrosis and leucocytic infiltration. Transport of Igs was seen in 77.8% cases of the first group and in all cases of groups 2 and 3. Transport of IgM was not found in the first group, but was seen in 50% cases of group 2 and 66.7% of group 3. PMID- 15822785 TI - [Morphological diagnosis and pathogenesis of non-developing pregnancy in antiphospholipid syndrome]. AB - 20 medical abortions and uterine scrapes from 24 females with verified antiphospholipid syndrome (APS) were studied by light and electron microscopy. Principle parts of pathogenesis are distinguished in the mother organism, target organs, placenta and uterine-placental region. Morphological criteria of APS diagnosis, in part histo-ultrastructural, pathology of microvilli of brush border, pathology of syncitiotrophoblast and villous cytotrophoblast. Rheological disturbances dominated in the parietal endometrium in the form of increasing hypercoagulative stratifying hemorrhages, thrombosis of uterine-placental arteries and microvessels, frequent formation of retroplacental hematoma. These structural straits are recommended for practical use in the work of pathologists. PMID- 15822786 TI - [Pathomorphology of adenohypophysis in cases of massive obstetric hemorrhages (morphometric and immunohistochemical studies)]. AB - There have been performed morphometric and immunohistochemical examinations of adenohypophysis of 15 females who died of massive post-partum hemorrhages. The revealed changes in the form of hypertrophy of the basophilic cells are the initial stages of Sheehan's syndrome developing as a result of long-term progression of hypopituitarism. Massive obstetric bleeding causes severe complications-posthemorrhagic shock, the pathogenetic mechanism of which includes necrosis of the cells in the anterior part of the hypophysis. Cause-effect relations were shown for the amount of the post-partum blood loss and the revealed pathomorphologic changes in the hypophysis. Hyperproduction of ACTH by the basophilic cells of the adenohypophysis revealed by immunohistochemical analysis is a compensatory mechanism under the conditions of acute adrenal insufficiency inherent in any shock including posthemorrhagic. PMID- 15822787 TI - [The role placental factors in formation of perinatal pathology in gestosis of different severity]. AB - Morphological study of gestosis in 153 cases. In light degree of gestosis focal ishemic and hemorrhagic infarcti and vascular changes of villi are found; decrease of the size of intervillous space at moderate degree and the 2nd stage of chronical placental hypertension. Hypoplasia of placentas, umbilical cord pathology and chronical placental hypertension in gestosis of a grave degree. PMID- 15822788 TI - [Morphologic and dopplerometric characteristics of spiral arteries in chronic placental insufficiency]. AB - Placental basal membrane of 156 females of the reproductive age with various forms of placental insufficiency was quantitatively studied. Dopplerometry of spiral arteries with calculation of the resistance index (RI) was performed longitudinally at 20-40 weeks of the pregnancy. Non-complete gestational restructurisation resulted in the decrease of circulation in these vessels with high values of RI and this is one of causes in delay of fetus development. High values of RI is an additional criterion of differential diagnosis of chronic placental insufficiency. PMID- 15822789 TI - [Morphological diagnosis of placental pathology in uterine contraction disorders]. AB - 100 placentas were studied and various forms of gestosis were diagnosed. Inflammation was frequent in weak activity of uterus. Discoordination of contractile uterus activity results in redistribution of fetal circulation resulting in vascular pathology of the umbilical cord and villi, in part gestation of vessels of terminal and intermedial villi and anemisation of trunk villi. PMID- 15822790 TI - [Gastritis associated with Helicobacter pylori in the material of gastrobiopsies in the group of children with dyspepsia]. AB - Helicobacter pylori (HP) was studied in gastrobiopsies from 180 children, 9 to 17 years old with chronic dyspeptic symptoms. The presence of HP was found in the stomach mucosa of 78.9% patients. The important morphological marker of HP infection was presence of lymphoid follicles and signs of "active" chronic gastritis in the stomach mucosa (p < 0.001). Thus HP may be considered as an essential pathogenetic factor of chronic dyspeptic syndrome in children of a school age. PMID- 15822791 TI - [Study of alpha1-antichymotrypsin (ACT) in prostatic carcinoma]. AB - 10 normal prostates and 100 prostates with tumour were studied immunohistochemically. ACT was found mainly in the cells lining the ducts. Synthesis of ACT is significantly increased in carcinoma due to mainly two parallel processes: ACT production by carcinoma cells and intensification of its production by normal cells mainly at the tumour periphery. Hyperplastic structures showed not very high ACT content, adenomatous structures revealed a higher response. On the whole, there is a parallelism between the content of ACT and prostatic specific antigen (PSA) in both normal and carcinomatous prostate, however PSA is being found in a much more wider spectrum of cells. Content of ACT in seminal fluid may be used as a parameter for diagnosis and monitoring of prostate cancer. PMID- 15822792 TI - [Alpha2-macroglobulin in various pathological conditions of the prostate]. AB - An immunohistochemical study of 10 normal prostates and 100 prostates with tumors showed the presence of alpha-2-macroglobulin (AMG)--an inhibitor of prostatic specific antigen (PSA)--in both normal organ and its tumor. In normal organ, AMG is located in the cells lining the ducts, while in the tumor tissue it is visualized in tubular structures and in parts of undifferentiated carcinoma. AMG is also found in stromal cells and in walls of vessels. Intensity of AMG immunohistochemical response in patients treated with antiandrogen in some cases increased. Intensity of reaction is decreasing when PSA concentration in the blood is very high. The authors suggest that AMG in the prostate is not only PSA inhibitor but also modulator of cell receptors influencing proliferative activity of tumour cells. PMID- 15822793 TI - [Prognostic value of structural-functional characteristics of skin melanoma]. AB - Surgical material is investigated. Morphometric criteria of ultrastructural atypia are considered as a possible prognostic factor of melanoma lymphogenic metastases. This allows detection of skin melanoma with high metastatic potential. PMID- 15822794 TI - [Generalized aspergillosis sepsis in a one month old infant with inborn leukemia]. AB - A rare case of aspergillesis sepsis in an one month old baby with inborn leukemia is reported. Of essential importance in the pathology is genetic deficiency of this child, decreased immunity and a grave intoxication due to leukemia chemotherapy. PMID- 15822795 TI - [Ganglioneuroma in a newborn with early inborn syphilis]. AB - A case is reported of a retroperitoneal sympathic nodes ganglioneuroma in a newborn with classic early inborn syphilis involving practically all systems of the viscera, skin, placenta. Some peculiar morphological features of the tumour are described. Possible teratogenic and oncogenic effects of treponema pallidus on embryonal fetal structures during intrauterine development are suggested. PMID- 15822796 TI - [Specialised pediatric pathology service]. AB - Leningrad Regional Children's Bureau of Pathology was established in May 1992. The program of this bureau consisted of: revealing causes of early and late abortions; assessment of ultrasound diagnosis of congenital deficiencies of fetus development and clinical assessment of chronic placental insufficiency; pathogenetic explanation of intrauterine fetuses death; prognosis of neonatal pathology; systemic analysis of dynamics of perinatal, natal and infant mortality; organisation of independant expertise of medical causes of reproductive losses. The following principles in the work of the bureau were established: 1) mass investigation of placentas of newborns; 2) use of urgent pathomorphological diagnosis which allows to present information about placentas not later than 24 hrs after the delivery; 3) complex investigation of placentas with the use of various methods; 4) systemic informational analysis of placental pathology; 5) constant updating of physicians knowledge in the field of practical placentology. Results of mass placentas investigation show a principal pathogenetic role of ascending infection in perinatal mortality in opportunistic pathogenic microflora with the development of inflammatory responses in the system mother-placenta-fetus. PMID- 15822797 TI - [The importance of specialised pathology service in diagnosis of ascending infection of the reproductive system in pregnant women]. AB - Practical results of screening cytological diagnosis of urogenital infections in pregnant women and mass pathomorphological investigations of newborn placentas performed by bureaus of pathology in Leningrad region and Cheboksary city are reviewed. Important place of the ascending bacterial infection in the structure of reproductive mortality is emphasized. PMID- 15822798 TI - [Independent expertise of medical service quality in perinatal, infantile and children's mortality]. AB - Information on principles and practical results of intraoffice expertise of medical documents in cases of the above mortality is presented. PMID- 15822799 TI - [Methods of separate heart section of fetuses and newborns]. AB - Modified Avtandilov's method of separate heart section of fetuses aged from 13 to 27 weeks of gestation obtained at artificial and spontaneous pregnancy interruption (newborn of 28 to 42 weeks of gestation) provided data of the atrial index (AI) and ventricular index (VI) in each age groups and distributed according to percentil method. Thus in the artificial fetuses normal AI is 1.118 1.353; VI-0.767-1.066; in spontaneous fetuses AI is 1.066-1.500; VI-0.772-1.083; in premature newborns AI is 1.116-1.538. Separation of heart compartments hypertrophy according to the degree is suggested. PMID- 15822800 TI - [Placentofetal relations as a basis for development and differentiation of definitive organs and tissues]. AB - On the basis of the analysis of clinical and experimental data a conclusion is made about existence of the regulatory-metabolic relations between different elements of the placenta and organs and tissues of the fetus which determine adaptive and pathological reactions in developing fetus. Regulation of metabolic relationships is a necessary part of systemogenesis and after development of the definitive cells and functional systems blocks the correlative systems. This mechanism is very important for development of the regulatory systems of the fetus forming postnatal homeostasis. PMID- 15822801 TI - Combined ultrasound and optoacoustic system for real-time high-contrast vascular imaging in vivo. AB - In optoacoustic imaging, short laser pulses irradiate highly scattering human tissue and adiabatically heat embedded absorbing structures, such as blood vessels, to generate ultrasound transients by means of the thermoelastic effect. We present an optoacoustic vascular imaging system that records these transients on the skin surface with an ultrasound transducer array and displays the images online. With a single laser pulse a complete optoacoustic B-mode image can be acquired. The optoacoustic system exploits the high intrinsic optical contrast of blood and provides high-contrast images without the need for contrast agents. The high spatial resolution of the system is determined by the acoustic propagation and is limited to the submillimeter range by our 7.5-MHz linear array transducer. A Q-switched alexandrite laser emitting short near-infrared laser pulses at a wavelength of 760 nm allows an imaging depth of a few centimeters. The system provides real-time images at frame-rates of 7.5 Hz and optionally displays the classically generated ultrasound image alongside the optoacoustic image. The functionality of the system was demonstrated in vivo on human finger, arm and leg. The proposed system combines the merits and most compelling features of optics and ultrasound in a single high-contrast vascular imaging device. PMID- 15822802 TI - Prospective motion correction of X-ray images for coronary interventions. AB - A method for prospective motion correction of X-ray imaging of the heart is presented. A 3D + t coronary model is reconstructed from a biplane coronary angiogram obtained during free breathing. The deformation field is parameterized by cardiac and respiratory phase, which enables the estimation of the state of the arteries at any phase of the cardiac-respiratory cycle. The motion of the three-dimensional (3-D) coronary model is projected onto the image planes and used to compute a dewarping function for motion correcting the images. The use of a 3-D coronary model facilitates motion correction of images acquired with the X ray system at arbitrary orientations. The performance of the algorithm was measured by tracking the motion of selected left coronary landmarks using a template matching cross-correlation. In three patients, we motion corrected the same images used to construct their 3D + t coronary model. In this best case scenario, the algorithm reduced the motion of the landmarks by 84%-85%, from mean RMS displacements of 12.8-14.6 pixels to 2.1-2.2 pixels. Prospective motion correction was tested in five patients by building the coronary model from one dataset, and correcting a second dataset. The patient's cardiac and respiratory phase are monitored and used to calculate the appropriate correction parameters. The results showed a 48%-63% reduction in the motion of the landmarks, from a mean RMS displacement of 11.5-13.6 pixels to 4.4-7.1 pixels. PMID- 15822803 TI - Operator dependence of 3-D ultrasound-based computational fluid dynamics for the carotid bifurcation. AB - The association between vascular wall shear stress (WSS) and the local development of atherosclerotic plaque makes estimation of in vivo WSS of considerable interest. Three-dimensional ultrasound (3DUS) combined with computational fluid dynamics (CFD) provides a potentially valuable tool for acquiring subject-specific WSS, but the interoperator and intraoperator variability associated with WSS calculations using this method is not known. Here, the accuracy, reproducibility and operator dependence of 3DUS-based computational fluid dynamics were examined through a phantom and in vivo studies. A carotid phantom was scanned and reconstructed by two operators. In the in vivo study, four operators scanned a healthy subject a total of 11 times, and their scan data were processed by three individuals. The study showed that with some basic training, operators could acquire accurate carotid geometry for flow reconstructions. The variability of measured cross-sectional area and predicted shear stress was 8.17% and 0.193 N/m2 respectively for the in vivo study. It was shown that the variability of the examined parameters was more dependent on the scan operators than the image processing operator. The range of variability of geometrical and flow parameters reported here can be used as a reference for future in vivo studies using the 3DUS-based CFD approach. PMID- 15822804 TI - Efficient pipeline for image-based patient-specific analysis of cerebral aneurysm hemodynamics: technique and sensitivity. AB - Hemodynamic factors are thought to be implicated in the progression and rupture of intracranial aneurysms. Current efforts aim to study the possible associations of hemodynamic characteristics such as complexity and stability of intra aneurysmal flow patterns, size and location of the region of flow impingement with the clinical history of aneurysmal rupture. However, there are no reliable methods for measuring blood flow patterns in vivo. In this paper, an efficient methodology for patient-specific modeling and characterization of the hemodynamics in cerebral aneurysms from medical images is described. A sensitivity analysis of the hemodynamic characteristics with respect to variations of several variables over the expected physiologic range of conditions is also presented. This sensitivity analysis shows that although changes in the velocity fields can be observed, the characterization of the intra-aneurysmal flow patterns is not altered when the mean input flow, the flow division, the viscosity model, or mesh resolution are changed. It was also found that the variable that has the greater impact on the computed flow fields is the geometry of the vascular structures. We conclude that with the proposed modeling pipeline clinical studies involving large numbers cerebral aneurysms are feasible. PMID- 15822805 TI - Efficient simulation of blood flow past complex endovascular devices using an adaptive embedding technique. AB - The simulation of blood flow past endovascular devices such as coils and stents is a challenging problem due to the complex geometry of the devices. Traditional unstructured grid computational fluid dynamics relies on the generation of finite element grids that conform to the boundary of the computational domain. However, the generation of such grids for patient-specific modeling of cerebral aneurysm treatment with coils or stents is extremely difficult and time consuming. This paper describes the application of an adaptive grid embedding technique previously developed for complex fluid structure interaction problems to the simulation of endovascular devices. A hybrid approach is used: the vessel walls are treated with body conforming grids and the endovascular devices with an adaptive mesh embedding technique. This methodology fits naturally in the framework of image-based computational fluid dynamics and opens the door for exploration of different therapeutic options and personalization of endovascular procedures. PMID- 15822806 TI - Segmentation of thrombus in abdominal aortic aneurysms from CTA with nonparametric statistical grey level appearance modeling. AB - This paper presents a new method for deformable model-based segmentation of lumen and thrombus in abdominal aortic aneurysms from computed tomography (CT) angiography (CTA) scans. First the lumen is segmented based on two positions indicated by the user, and subsequently the resulting surface is used to initialize the automated thrombus segmentation method. For the lumen, the image derived deformation term is based on a simple grey level model (two thresholds). For the more complex problem of thrombus segmentation, a grey level modeling approach with a nonparametric pattern classification technique is used, namely k nearest neighbors. The intensity profile sampled along the surface normal is used as classification feature. Manual segmentations are used for training the classifier: samples are collected inside, outside, and at the given boundary positions. The deformation is steered by the most likely class corresponding to the intensity profile at each vertex on the surface. A parameter optimization study is conducted, followed by experiments to assess the overall segmentation quality and the robustness of results against variation in user input. Results obtained in a study of 17 patients show that the agreement with respect to manual segmentations is comparable to previous values reported in the literature, with considerable less user interaction. PMID- 15822807 TI - Vessel tree reconstruction in thoracic CT scans with application to nodule detection. AB - Vessel tree reconstruction in volumetric data is a necessary prerequisite in various medical imaging applications. Specifically, when considering the application of automated lung nodule detection in thoracic computed tomography (CT) scans, vessel trees can be used to resolve local ambiguities based on global considerations and so improve the performance of nodule detection algorithms. In this study, a novel approach to vessel tree reconstruction and its application to nodule detection in thoracic CT scans was developed by using correlation-based enhancement filters and a fuzzy shape representation of the data. The proposed correlation-based enhancement filters depend on first-order partial derivatives and so are less sensitive to noise compared with Hessian-based filters. Additionally, multiple sets of eigenvalues are used so that a distinction between nodules and vessel junctions becomes possible. The proposed fuzzy shape representation is based on regulated morphological operations that are less sensitive to noise. Consequently, the vessel tree reconstruction algorithm can accommodate vessel bifurcation and discontinuities. A quantitative performance evaluation of the enhancement filters and of the vessel tree reconstruction algorithm was performed. Moreover, the proposed vessel tree reconstruction algorithm reduced the number of false positives generated by an existing nodule detection algorithm by 38%. PMID- 15822808 TI - Novel approaches to the measurement of arterial blood flow from dynamic digital X ray images. AB - We have developed two new algorithms for the measurement of blood flow from dynamic X-ray angiographic images. Both algorithms aim to improve on existing techniques. First, a model-based (MB) algorithm is used to constrain the concentration-distance curve matching approach. Second, a weighted optical flow algorithm (OP) is used to improve on point-based optical flow methods by averaging velocity estimates along a vessel with weighting based on the magnitude of the spatial derivative. The OP algorithm was validated using a computer simulation of pulsatile blood flow. Both the OP and the MB algorithms were validated using a physiological blood flow circuit. Dynamic biplane digital X-ray images were acquired following injection of iodine contrast medium into a variety of simulated arterial vessels. The image data were analyzed using our integrated angiographic analysis software SARA to give blood flow waveforms using the MB and OP algorithms. These waveforms were compared to flow measured using an electromagnetic flow meter (EMF). In total 4935 instantaneous measurements of flow were made and compared to the EMF recordings. It was found that the new algorithms showed low measurement bias and narrow limits of agreement and also out-performed the concentration-distance curve matching algorithm (ORG) and a modification of this algorithm (PA) in all studies. PMID- 15822809 TI - Assessment of vulnerable plaque composition by matching the deformation of a parametric plaque model to measured plaque deformation. AB - Intravascular ultrasound (IVUS) elastography visualizes local radial strain of arteries in so-called elastograms to detect rupture-prone plaques. However, due to the unknown arterial stress distribution these elastograms cannot be directly interpreted as a morphology and material composition image. To overcome this limitation we have developed a method that reconstructs a Young's modulus image from an elastogram. This method is especially suited for thin-cap fibroatheromas (TCFAs), i.e., plaques with a media region containing a lipid pool covered by a cap. Reconstruction is done by a minimization algorithm that matches the strain image output, calculated with a parametric finite element model (PFEM) representation of a TCFA, to an elastogram by iteratively updating the PFEM geometry and material parameters. These geometry parameters delineate the TCFA media, lipid pool and cap regions by circles. The material parameter for each region is a Young's modulus, EM, EL, and EC, respectively. The method was successfully tested on computer-simulated TCFAs (n = 2), one defined by circles, the other by tracing TCFA histology, and additionally on a physical phantom (n = 1) having a stiff wall (measured EM = 16.8 kPa) with an eccentric soft region (measured EL = 4.2 kPa). Finally, it was applied on human coronary plaques in vitro (n = 1) and in vivo (n = 1). The corresponding simulated and measured elastograms of these plaques showed radial strain values from 0% up to 2% at a pressure differential of 20, 20, 1, 20, and 1 mmHg respectively. The used/reconstructed Young's moduli [kPa] were for the circular plaque EL = 50/66, EM = 1500/1484, EC = 2000/2047, for the traced plaque EL = 25/1, EM = 1000/1148, EC = 1500/1491, for the phantom EL = 4.2/4 kPa, EM = 16.8/16, for the in vitro plaque EL = n.a./29, EM = n.a./647, EC = n.a./1784 kPa and for the in vivo plaque EL = n.a./2, EM = n.a./188, Ec = n.a./188 kPa. PMID- 15822810 TI - Geometric modeling of the human normal cerebral arterial system. AB - We propose an anatomy-based approach for an efficient construction of a three dimensional human normal cerebral arterial model from segmented and skeletonized angiographic data. The centerline-based model is used for an accurate angiographic data representation. A vascular tree is represented by tubular segments and bifurcations whose construction takes into account vascular anatomy. A bifurcation is defined quantitatively and the algorithm calculating it is given. The centerline is smoothed by means of a sliding average filter. As the vessel radius is sensitive to quality of data as well as accuracy of segmentation and skeletonization, radius outlier removal and radius regression algorithms are formulated and applied. In this way, the approach compensates for some inaccuracies introduced during segmentation and skeletonization. To create the frame of vasculature, we use two different topologies: tubular and B-subdivision based. We also propose a technique to prevent vessel twisting. The analysis of the vascular model is done on a variety of data containing 258 vascular segments and 131 bifurcations. Our approach gives acceptable results from anatomical, topological and geometrical standpoints as well as provides fast visualization and manipulation of the model. The approach is applicable for building a reference cerebrovascular atlas, developing applications for simulation and planning of interventional radiology procedures and vascular surgery, and in education. PMID- 15822811 TI - Visualization of vasculature with convolution surfaces: method, validation and evaluation. AB - We present a method for visualizing vasculature based on clinical computed tomography or magnetic resonance data. The vessel skeleton as well as the diameter information per voxel serve as input. Our method adheres to these data, while producing smooth transitions at branchings and closed, rounded ends by means of convolution surfaces. We examine the filter design with respect to irritating bulges, unwanted blending and the correct visualization of the vessel diameter. The method has been applied to a large variety of anatomic trees. We discuss the validation of the method by means of a comparison to other visualization methods. Surface distance measures are carried out to perform a quantitative validation. Furthermore, we present the evaluation of the method which has been accomplished on the basis of a survey by 11 radiologists and surgeons. PMID- 15822812 TI - Robust quantification of in vitro angiogenesis through image analysis. AB - An automated image analysis method for quantification of in vitro angiogenesis is presented. The method is designed for in vitro angiogenesis assays that are based on co-culturing endothelial cells with fibroblasts. Such assays are used in many current studies in which anti-angiogenic agents for the treatment of cancer are being sought. This search requires accurate quantification of the stimulatory and inhibitory effects of the different agents. The quantification method gives lengths and sizes of the tubule complexes as well as the numbers of junctions in each of them. The method is tested with a set of test images obtained with a commercially available in vitro angiogenesis assay. The results correctly indicate the inhibitory effect of suramin and the stimulatory effect of vascular endothelial growth factor. Moreover, the image analysis method is shown to be robust against variations in illumination. We have implemented a software package that utilizes the methods. The software as well as a set of test images are available at http://www.cs.tut.fi/sgn/csb/angioquant/. PMID- 15822813 TI - Genetic pathways to glioblastomas. AB - Glioblastomas, the most frequent and malignant human brain tumors, may develop de novo (primary glioblastoma) or by progression from low-grade or anaplastic astrocytoma (secondary glioblastoma). These glioblastoma subtypes constitute distinct disease entities that affect patients of different ages and develop through different genetic pathways. Our recent population-based study in the Canton of Zurich, Switzerland, shows that primary glioblastomas develop in older patients (mean age, 62 years) and typically show LOH on chromosome 10q (69%) and other genetic alterations (EGFR amplification, TP53 mutations, p16INK4a deletion, and PTEN mutations) at frequencies of 24-34%. Secondary glioblastomas develop in younger patients (mean, 45 years) and frequently show TP53 mutations (65%) and LOH 10q (63%). Common to both primary and secondary glioblastoma is LOH on 10q, distal to the PTEN locus; a putative suppressor gene at 10q25-qter may be responsible for the glioblastoma phenotype. Of the TP53 point mutations in secondary glioblastomas, 57% were located in hotspot codons 248 and 273, while in primary glioblastomas, mutations were more widely distributed. Furthermore, G:C- >A:T mutations at CpG sites were more frequent in secondary than in primary glioblastomas (56% vs 30%). These data suggest that the TP53 mutations in these glioblastoma subtypes arise through different mechanisms. There is evidence that G:C-->A:T transition mutations at CpG sites in the TP53 gene are significantly more frequent in low-grade astrocytomas with promoter methylation of the O6 methylguanine-DNA methyltransferase (MGMT) gene than in those without methylation. This suggests that, in addition to deamination of 5-methylcytosine (the best known mechanism of formation of G:C-->A:T transitions at CpG sites), involvement of alkylating agents that produce O6-methylguanine or related adducts recognized by MGMT cannot be excluded in the pathway leading to secondary glioblastomas. PMID- 15822814 TI - Mechanisms of face perception in humans: a magneto- and electro-encephalographic study. AB - We have been studying the underlying mechanisms of face perception in humans using magneto- (MEG) and electro-encephalography (EEG) including (1) perception by viewing the static face, (2) differences in perception by viewing the eyes and whole face, (3) the face inversion effect, (4) the effect of gaze direction, (5) perception of eye motion, (6) perception of mouth motion, and (7) the interaction between auditory and visual stimuli related to the vowel sounds. In this review article, we mainly summarize our results obtained on 3, 5, and 6 above. With the presentation of both upright and inverted unfamiliar faces, the inferior temporal cortex (IT) centered on the fusiform gyrus, and the lateral temporal cortex (LT) near the superior temporal sulcus were activated simultaneously, but independently, between 140 and 200 ms post-stimulus. The right hemisphere IT and LT were both active in all subjects, and those in the left hemisphere in half of the subjects. Latencies with inverted faces relative to those with upright faces were longer in the right hemisphere, and shorter in the left hemisphere. Since the activated regions under upright and those under inverted face stimuli did not show a significant difference, we consider that differences in processing upright versus inverted faces are attributable to temporal processing differences rather than to processing of information by different brain regions. When viewing the motion of the mouth and eyes, a large clear MEG component, 1M (mean peak latency of approximately 160 ms), was elicited to both mouth and eye movement, and was generated mainly in the occipito-temporal border, at human MT/V5. The 1M to mouth movement and the 1M to eye movement showed no significant difference in amplitude or generator location. Therefore, our results indicate that human MT/V5 is active in the perception of both mouth and eye motion, and that the perception of movement of facial parts is probably processed similarly. PMID- 15822815 TI - Blood tumor permeability of experimental brain tumor: an electron microscopic study using lanthanum. AB - In an attempt to assess the permeability of microvessels in the experimental brain tumor model, lanthanum ion (La3+) was used as a low-molecular weight electron microscopic probe. Rat glioma 9 L and adenocarcinoma ACL15 were transplanted to the brain and subflank of rats. The rats were then anesthetized sequentially perfused with saline, saline plus La3+ followed by a fixative in phosphate buffer. The brain and subcutaneous tumors were removed, further fixed, and processed for electron microscopy. La3 did not pass through the tight junctions of the normal cerebral endothelium. Similarly, La3+ did not penetrate the endothelial cell wall of the microvessels in the transplanted brain tumors. In contrast, extravasation of La3+ from the microvessels in the transplanted subcutaneous tumors was observed. The electron microscopy examination results indicate that the vesicular transport was a predominant mechanism in the penetration of La3+ through the endothelial cell wall. Since most chemotherapeutic agents similar as La3+ are of low molecular weight, we can suggest from the results of our present study that the blood tumor permeability of the anti-cancer agents in the rat model of brain glioma transplantation differs from that in the rat model of subcutaneous glioma transportation. In other words, our results indicate that when the subcutaneous glioma transplantation model is used in sensitivity tests of anti-cancer agents, it will possibly be very difficult to predict the anti-neoplastic effect in vivo. PMID- 15822816 TI - Glyoxal inactivates glutamate transporter-1 in cultured rat astrocytes. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor paralysis and selective motor neuron death. There is increasing evidence that motor neuron death in ALS is mediated by glutamate toxicity resulting from reduced activity of astrocytic glutamate transporter-1 (GLT-1). Recent morphological studies have shown that Nepsilon (carboxymethyl)lysine (CML) accumulates in reactive astrocytes of ALS spinal cords. CML is a product of post-translational protein modification by glyoxal, a reactive aldehydic intermediate. In considering these documents, it is important to determine whether GLT-1 protein modification by glyoxal might cause reduced GLT-1 activity. To address this issue, we investigated the effects of glyoxal on GLT-1 properties in cultured rat astrocytes. High performance liquid chromatography showed reduced glutamate uptake activity in the glyoxal-exposed cells. Immunocytochemical analysis displayed CML accumulation in the cytoplasm of astrocytes by glyoxal exposure. Immunoblots of immunoprecipitated GLT-1 disclosed GLT-1 CML adduct formation in the glyoxal-exposed cells. Our results indicate that glyoxal modifies GLT-1 to form CML and simultaneously deprives its glutamate uptake activity. Thus, these toxic effects of glyoxal on astrocytes might be implicated in motor neuron death in ALS. PMID- 15822817 TI - Forced retraction of spinal root injury enhances activation of p38 MAPK cascade in infiltrating macrophages. AB - Root-rupture injury is a type of preganglionic brachial plexus injury resulting from traction force, where a small section of the spinal root is usually left behind. We have established experimental models of both root-rupture injury with traction force and rhizotomy without traction force in rats and we examined the activation of microglia/ macrophages in both conditions. LGP107 and LGP96, which are rat homologs of lysosome-associated membrane proteins, were most useful as immunohistochemical markers of mononuclear phagocytes. The metabolic activation of macrophages was analyzed by immunohistochemistry with a series of antibodies against tumor necrosis factor-alpha (TNF-alpha), cathepsin B, p38 mitogen activated protein kinase (MAPK), and mitogen-activated kinase kinase 3 (MKK3). Both root-rupture injury and rhizotomy rapidly induced the aggregation of numerous macrophages from the injured dorsal root to the dorsal funiculus and TNF alpha was highly expressed by the macrophages in the injured dorsal root at 48 h. Activation of p38 MAPK was preferentially observed in the macrophages at the ruptured dorsal root; however, only slight activation of p38 MAPK was observed at the rhizotomized dorsal root. These findings suggest that traction injury of the spinal root might induce activation of the p38 MAPK cascade and production of TNF alpha in the infiltrating macrophages, both of which might participate in aggravation of the root injury. PMID- 15822818 TI - Comparative status of activated ERK1/2 and PARP cleavage in human gliomas. AB - Gliomas are the most common form of cerebral tumors. Understanding molecular features of glioma will eventually allow for targeted intervention and more promising approaches for treating gliomas. The present study is therefore carried out to check the levels of activated ERK1/2 with respect to phospho-tyrosine and cleavage of poly ADP-ribose polymerase (PARP). Recent experiments support that extracellular signal regulated kinase (ERK), a mitogen activated protein (MAP) kinase might have a critical role in cell proliferation. PARP is a DNA-repair enzyme activated by DNA strand breaks. Overactivation of PARP after cellular insult lead to cell death caused by rapid depletion of cellular ATP. Three glioblastoma multiforme (GBM) and two astrocytoma biopsies (core tumor) and peripheral tissues were analyzed for the expression of p-ERK1/2 and PARP. Results indicate higher p-ERK1/2 in GBM. Cleaved fragments of PARP (89 kDa) were found to be more in core tumor tissue samples as compared to peripheral tumor tissues of both astrocytoma and GBM. PMID- 15822819 TI - Increase of Cardiotrophin-1 immunoreactivity in regenerating and overloaded but not denervated muscles of rats. AB - The original report by Pennica et al. on Cardiotrophin-1 (CT-1) states that it markedly stimulates hypertrophy in cardiac myocytes both in vitro and in vivo and is predominantly expressed in the early mouse embryonic heart tube. CT-1 is a member of the interleukin-6 superfamily and past studies have shown that it exerts trophic effects on neurons, glial cells and their precursors, and is expressed during myogenesis. Thus CT-1 is associated with physical and pathological changes in skeletal muscle. In this study, we examined whether CT-1 is expressed in mechanically overloaded, regenerating, and denervated muscles of rats using immunohistochemistry. In the overloaded plantaris muscles at 1 and 3 days postsurgery, CT-1 immunoreactivity was detected in the mononuclear cells that had infiltrated the extracellular space. CT-1 immunoreactivity was also observed in the mononuclear cells invading the extracellular space at 2, 4, and 6 days after a bupivacaine injection and in degenerative and necrotic muscle fibers at 2 days postinjection. In the denervated muscles, the CT-1 immunoreactivity did not change in intensity during the entire period of the denervation (2, 7, and 14 days postsurgery). The cells invading extracellular space and in necrotic muscle fibers possessing CT-1 immunoreactivity might be muscle precursor cells (satellite cells) or migrating macrophages undergoing phagocytosis. Using double immunostainings for anti-CT-1/antic-met, anti-CT-1/ anti-M-cadherin, and anti-CT 1/anti-ED1, we found that satellite cells and macrophages exhibited CT-1 immunoreactivity in the damaged muscles after bupivacaine injection. We therefore believe that CT-1 plays a key role in regeneration and hypertrophy in the skeletal muscle of rats. PMID- 15822820 TI - A novel coating biomaterial for intracranial aneurysms: effects and safety in extra- and intracranial carotid artery. AB - Methyl-2-cyanoacrylate, a widely used material for coating cerebral aneurysm, was recently withdrawn. The aim of the present study was to develop an alternative coating material for cerebral aneurysm, which is safe, effective and stable within the brain. In the first experiment, an aneurysm model of the common carotid artery was produced in a rabbit by the local application of elastase. The aneurysm produced was covered by no material (Group A), a cellulose cotton sheet and conventional methyl-2-cyanoacrylate (Group B), a newly produced polyglycolic acid felt and fibrin glue (Group C), or a cellulose cotton sheet and fibrin glue (Group D). Histological examination showed that the materials resulted in the formation of tight connective tissue around the artery, and that the material was completely replaced by the connective tissue after 12 weeks. This change was found exclusively in Group C, but not in Group A or the other materials, although a temporary thickening of the intima was also observed at the site of the elastase application in Group C. In Group D, a long-term, marked thickening of the intima was observed. In the second experiment, using an intracranial internal carotid artery from a beagle, the applied polyglycolic acid felt and fibrin glue to the intracranial artery induced the formation of connective tissue around the artery that was completely absorbed 16 weeks after surgery. There were no signs of intimal thickening or of adverse reactions in nervous tissue. The present results suggest that polyglycolic acid felt and fibrin glue is a possible candidate for a safe, effective biomaterial to wrap or coat cerebral aneurysm. PMID- 15822821 TI - Cerebellar liponeurocytoma: morphological, immunohistochemical, and ultrastructural study of a relapsed case. AB - Cerebellar liponeurocytoma is a rare and newly identified neoplasm found in adults which is reputed to be benign. Its salient morphological characteristics are advanced neuronal/neurocytic differentiation, the presence of lipomatous areas, low mitotic rate, and the absence of necrosis, pleomorphism and vascular hyperplasia. Reported is a case of relapsing liponeurocytoma which occurred 3 and a half years after the radical excision of the primary lesion. Histopathological aggressive features (mitoses and a high proliferation index as evaluated by MIB 1) were shown in the primary lesion and recurrence of the tumor. We suggest that liponeurocytoma is an uncertain malignant potential lesion when mitoses are present and the MIB-1 positive cells are more than 10%. PMID- 15822822 TI - Chondromyxoid fibroma of the parietal bone. AB - We report the case of an 11-year old girl who presented with an intracranial chondromyxoid fibroma arising from parietal bone. Chondromyxoid fibroma is a benign cartilaginous neoplasm, which can be easily mistaken for chondrosarcoma, especially in small biopsies. It is more common in long bones and is least common in cranial bones. Published work describes about 33 cases occurring in cranial bones with almost equal incidence in the bones of the base of the skull and cranial vault. To our knowledge, this is the second case of chondromyxoid fibroma reported to occur in parietal bone and is the first case reported in an English language journal. We present this case for its rare anatomical, clinical, radiological and histological presentations. PMID- 15822823 TI - Leptomeningeal carcinomatosis from urinary bladder adenocarcinoma: a clinicopathological case study. AB - We report a 73-year-old male patient with leptomeningeal metastasis from urinary bladder adenocarcinoma. He was presented with prominent hyperactive delirium during the course of the disease. Meningeal carcinomatosis was detected 5 days before his death, but the primary site of the malignant tumor could not be determined. Necropsy revealed leptomeningeal infiltration of many adenocarcinoma cells that covered the cerebrum. The leptomeninges of the right middle frontal gyrus, superior temporal gyrus, precentral gyrus and inferior parietal lobe were most severely affected by tumor cell infiltration. Cerebral edema was found to extensively cover the basal part of the temporal lobe. In the cerebrum, tumor cells were clustered in the perivascular spaces and had invaded localized areas of the frontal lobe. Vascular cell adhesion molecule (VCAM)-1 expression was detected in the small vessels of the cerebral upper cortical layers and of temporal subcortical u-fibers. Numerous astrocytes positive for cytokeratin AE1/AE3 were found in the frontal and temporal lobes. Meningeal carcinomatosis from urinary bladder adenocarcinoma is extremely rare and up-regulation of the adhesion molecules in the meningeal adenocarcinoma was confirmed. PMID- 15822824 TI - Peculiar venous lesions in fatal hyponatremic brain edema. AB - A 19-year-old woman with a 3-year history of schizophrenia suddenly began to vomit, and rapidly developed a coma an hour after the onset of vomiting. A brain CT scan showed diffuse brain edema with compression of the ventricles. Laboratory tests showed a low serum sodium concentration of 117 mmol/L. She died 67 h after the onset of the first symptom. A postmortem examination showed diffuse swelling of the brain with bilateral uncal and tonsillar herniations. Histologically, no necrotic, hemorrhagic or encephalitic changes were seen. However, microvacuolar changes with lymphocytic infiltration were found in the venous walls (media and adventitia) mainly in the basal ganglia, thalamus and brainstem. To our knowledge, this is the first demonstration of venous alterations in fatal hyponatremic brain edema. These changes may have participated in the exacerbation of the brain edema due to functional disturbance of venous drainage. PMID- 15822825 TI - Chronic calcified subdural empyema occurring 46 years after surgery. AB - The authors present a case of a 47-year-old female in whom there was diagnosed a chronic calcified subdural empyema 46 years after the removal of an acute subdural empyema resulting from complications after otitis media. The patient had suffered from grand mal convulsions and partial epileptic seizures occurring 3-4 times a month. A large frontotemporoparietal craniotomy was carried out and the subdural empyema filled with numerous brownish-black, uncharacteristic tissue fragments together with the partially calcified and ossified capsule was removed. The empyema mass was found to be sterile for bacteria. After the operation, mental disability symptoms began to withdraw and the number of epileptic seizures decreased. PMID- 15822826 TI - A 35-year-old female with growth and developmental retardation, progressive ataxia, dementia and visual loss. PMID- 15822827 TI - A 74-year-old woman presenting slowly progressive dementia and emotional instability. PMID- 15822828 TI - Improving health, eliminating disparities: finding solutions for better health care for all populations. AB - The Fourth Annual Primary Care and Prevention Conference was held October 25-27, 2004 in Atlanta, Georgia to address inequalities in primary care health outcomes and to develop strategies to improve the health status among under-served communities throughout the country. The Ninth Annual HeLa Women's Health Conference was conducted concurrently during the conference's Tuesday, October 26 sessions and were designed for those in obstetrics/gynecology. The reports herein provide a sample of the rich presentations and scientific knowledge imparted by a faculty of more than 100 who addressed disparities across disease states (heart disease and stroke; cancer; diabetes, chronic kidney disease, and obesity; infectious disease; mental health and mental disorders; environmental health/injury and violence; respiratory diseases; and substance abuse, tobacco use) and within focused areas (adult health; maternal, child and adolescent health; health policy and practice; and social and community health). PMID- 15822829 TI - Racial and ethnic disparities in breast cancer stage, treatment, and survival in the United States. AB - Several studies indicate that African-American, Hispanic White, and Native American women with breast cancer present with more advanced stages and have poorer survival rates than non-Hispanic Whites, while Asians/Pacific Islanders do not. However, Asians/Pacific Islanders and Hispanic Whites are heterogeneous populations, and recent data indicate that certain subgroups of these populations have poorer breast cancer outcomes compared to non-Hispanic Whites, while others have better outcomes. Many of these disparities have persisted for decades, but until recently, detailed studies exploring the reasons behind these disparities have been limited. The results of these studies point to the effect of differences in socioeconomic status, access to health care (including both breast cancer screening and treatment services), lifestyle factors, and tumor characteristics on these disparities. Thus, these studies indicate that these disparities are multifactorial, and therefore strategies aimed at reducing them must involve advocacy, research, education, and healthcare services. A key component to the success of these strategies is not only support for them on the federal and state levels, but also the involvement of local communities in developing programs and policies that are culturally and linguistically appropriate for their communities in order to ensure not only the utility, but also the longevity, of these efforts. PMID- 15822830 TI - Smoking cessation: what works in primary care settings. AB - This paper presents evidence-based strategies for assisting patients in quitting smoking. Summaries of the effectiveness of advice alone, varying levels of behavioral counseling, nicotine replacement therapies, and bupropion either alone or in combinations are presented. We describe two web-based resources, one that offers guidance to practitioners and another that is intended as a training tool for students or healthcare professionals on ways to intervene with patients who smoke. The training curriculum contains both didactic material based on the U.S. Public Health Service Clinical Practice Guideline and tutorials with simulated patients depicting various types of real-life scenarios. The curriculum can meet the need for training among both students and practitioners in medicine, nursing, health education, dentistry, and other healthcare professions. PMID- 15822831 TI - Community health advisors effectively promote cancer screening. AB - Cancer is the second leading cause of death in the United States. Across many cancers, racial and ethnic minorities bear an unequal burden of disease (e.g., lower 5-year survival rates). Concerted efforts to reach minority communities and address these disparities are needed. Community Health Advisors (CHAs) are trusted natural helpers from within the community who provide emotional support, advice, and tangible aid to members of their social network. Their understanding of community culture allows them to provide culturally appropriate, informal, and spontaneous assistance to community members. Using various approaches and across a number of racial/ethnic groups, CHA interventions have demonstrated success in increasing cancer screening. This paper highlights some of those intervention programs and concludes with considerations for future projects. PMID- 15822832 TI - Utilization of services among elderly cancer patients--relationship to age, symptoms, physical functioning, comorbidity, and survival status. AB - In this study we investigated predictors of utilization of primary care physician, hospital and emergency room services in a sample of 909 older patients during the first year following a diagnosis of cancer of the breast, colon, lung, or prostate. Analysis of covariance models were implemented separately for the active treatment period (0-6 months) and the continuing care period (6-12 months) to determine how age, sex, comorbidity, length of survival, treatment status, stage of disease, cancer site, physical functioning, and symptom count were related to primary care physician visits, hospitalization, and emergency room use. Decreased physical functioning was related to increased physician visits, hospital nights, and emergency room visits during the active treatment period, and to increased hospital nights and emergency room visits during the continuing care period. Patients with three or more comorbid conditions reported more physician visits than patients with no comorbid conditions during both periods. Patient age did not play a significant role in utilization of services. The broad picture suggested by this study of elderly cancer patients is that their service utilization, particularly hospitalization and emergency room services, tends to peak in concert with a dramatic decrease in physical functioning as the patient nears the end of life. Use of primary care physicians' services may depend substantially on comorbid conditions. PMID- 15822833 TI - Improving cardiovascular health outcomes through the use of evidence-based medicine. AB - The risk of cardiovascular (CV) and renal complications begins at a relatively low blood pressure (BP), and this risk is associated with such disorders as metabolic syndrome. When comparing older antihypertensive agents with newer agents, for the most part no significant differences have been seen in rates of CV events. However, rapidly controlling BP is critical to reduce event rates, particularly rates of stroke. Angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers consistently decrease the rate of onset of type 2 diabetes. PMID- 15822834 TI - Health care and civil rights: an introduction. AB - This article offers a brief history of healthcare civil rights, describes a range of healthcare issues that have a civil rights component, and discusses the need for an expanded civil rights framework to guide the provision of health care. Unequal health care based on race and ethnicity has received renewed attention over the past several years, but healthcare discrimination based on socioeconomic status, disability, age, and gender also deserve careful attention. PMID- 15822835 TI - Racial disparities and site of care. AB - Numerous studies have demonstrated that minority patients receive poorer quality health care than non-minorities. The mechanisms underlying this problem have not been identified, but the pervasiveness and consistency of racial and ethnic differences in healthcare quality have led most investigators to identify at most one or two overarching causes. To some, the consistency of these findings supports a hypothesis that physicians are at the heart of the problem. It is posited that due to sub-conscious biases, more overt prejudice, or cultural insensitivity, physicians do not treat minority patients as well as they treat non-minority patients. This hypothesis has received a great deal of attention, both in reviews from the Institute of Medicine and position statements from the American Medical Association and National Medical Association. In this paper, I review several studies that have focused on an alternative potential mechanism of racial and ethnic disparities in health care, which is based more on inequities in the structure of the healthcare system, rather than inequities in the treatment patterns of individual physicians. Determining the relative contribution of each of these mechanisms to racial and ethnic disparities in health care should be a priority. PMID- 15822836 TI - Post menopausal hormone therapy today. PMID- 15822837 TI - The Ebony Sex Survey and the sex lives of African-American women: a call to healthcare providers. AB - Abuse, sexual satisfaction, and sexual dysfunction are issues that are not readily addressed in the African-American population, but they are on the minds of many. Ebony decided to pose these and other questions to African-American women in a survey titled, "Are You Satisfied?" Respondents were subscribers to Ebony as well as newsstand readership. Participants either filled out the survey on the Ebony website or sent in a paper copy to be tabulated. African-American women (N=7800) from all walks of life responded and discussed their abuse histories, their relationships, their ability or inability to reach orgasm, their medical histories and level of comfort in discussing their problems with their physicians, as well as many other issues. Results indicate that 37% of the sample self-reported early sexual abuse at the hands of immediate and/or extended family members; 15%-18% have been treated for high blood pressure, STDs/HIV, and/or fibroids; and 33% have experienced an inability to achieve orgasm in the last year. Of the sample 47% stated that they had not discussed these problems with their physicians. PMID- 15822838 TI - Interstitial lung disease: causes, treatment, and prevention. AB - The interstitial lung diseases (ILD) are a diverse group of lung disorders that involve primarily the parenchyma of the lung. Whether idiopathic or secondary to systemic disorders, inhaling exposures, or drugs, they inflame and scar the interstitium of the lungs and obliterate alveoli and capillary units. The scarring or fibrosis produces restrictive lung impairment while destruction of the alveoli, interstitium, and capillaries results in severe gas exchange abnormalities. Clinically, the ILD present subtly with progressive dyspnea on exertion and a dry cough. Rales or crackles on examination prompt chest radiography that may reveal bilateral infiltrates. These infiltrates are often treated as atypical pneumonias that fail to respond to antimicrobial therapy over weeks to months. Because of this and their infrequent presentation in the primary care setting, the diagnosis of ILD is commonly delayed. This paper highlights the natural history of the ILD in general, gives a broad overview of the pathophysiology in these diseases, and encourages greater awareness for the detection of ILD in primary care. PMID- 15822839 TI - Metro Atlanta responds to West Nile virus: a coordinated public health response. AB - Three and a half million people live in metropolitan Atlanta, in multiple counties with varying population bases, resources, issues and separate boards of health. Historically, public health issues have been managed within each county, with very little sharing of information among counties. The 1999 West Nile virus (WNV) outbreak in the Northeast caused public health officials in Atlanta to recognize the potential for the disease to spread to Georgia and the need to develop a coordinated, multi-jurisdictional response plan. This plan would need to address a new disease with little scientific data to predict how it might behave in a new environment and would also require closely coordinated communication among the local/state public health entities and elected officials. In early 2000, staff from the five health districts in the metro Atlanta area and the state health department voluntarily convened the Metro Atlanta Surveillance Task Force (MASTF) to create the Metro Atlanta West Nile Virus Response Plan. This plan utilizes a coordinated effort encompassing public education, surveillance, and mosquito control. With this plan in place, when the first human case of WNV was detected in Atlanta, the public heard consistent health messages about preventive measures to lower their risk of illness and the metro counties were able to carry out a successful uniform approach to mosquito control. This plan has received recognition by the National Association of County and City Health Officials (NACCHO) as a 2004 Model Practice, demonstrating exemplary and replicable qualities in response to a local public health need. Since the early days of the emergence of WNV in the metro Atlanta area, MASTF has continued to be a viable, evolving entity, managing and anticipating health issues. The MASTF plan is a successful effort to develop consistent policies and procedures for disease surveillance in a heavily populated area with multiple local health departments. PMID- 15822840 TI - The hepatitis C crisis. AB - An estimated 170 million persons (3% of the world's population) are infected with the hepatitis C virus (HCV), and 3 to 4 million persons are newly infected each year. Of those infected, 70%-85% develop chronic viremia with the potential for devastating long-term sequelae, including chronic liver disease, cirrhosis, hepatic failure, and hepatocellular carcinoma. The passivity in the public health sector and in the medical community at large with respect to hepatitis C portends a myriad of societal, fiscal, and personal costs for the United States within the next two decades unless immediate actions are taken to intervene in the natural history of this emerging public health crisis. PMID- 15822841 TI - Plenary session 1: A Surgeons General colloquium. Major health policy challenges and opportunities facing the nation. PMID- 15822842 TI - Plenary session 1A: Primary healthcare challenges. Preventing and treating disease. PMID- 15822843 TI - Plenary session 2: Improving quality. Making a difference at the individual, business, and community levels. PMID- 15822844 TI - Plenary session 3: Health policy 2004. Improving outcomes through research and practice. PMID- 15822845 TI - Plenary session 4: Improving health, eliminating disparities: visions for the 21st century. PMID- 15822847 TI - Governing the captives: forensic psychiatric nursing in corrections. AB - TOPIC/PROBLEM: Since 1978, the federal inmates of Canada serving time have had access to a full range of psychiatric care within the carceral system. Five psychiatric units are part of the Federal Correctional Services. Nursing practice in forensic psychiatry opens up new horizons in nursing. This complex professional nursing practice involves the coupling of two contradictory socio professional mandates: to punish and to provide care. METHOD: The purpose of this article is to present the results of a grounded theory doctoral study realized in a multi-level security psychiatric ward of the Canadian Federal Penitentiary System. The theoretical work of the late French philosopher, Michel Foucault, and those of sociologist, Erving Goffman, are used to illuminate the qualitative data that emerged from the author's fieldwork. FINDINGS: A Foucauldian perspective allows us to understand the way forensic psychiatric nursing is involved in the governance of mentally ill criminals through a vast array of power techniques (sovereign, disciplinary, and pastoral) which posited nurses as "subjects of power". These nurses are also "objects of power" in that nursing practice is constrained by formal and informal regulations of the penitentiary context. CONCLUSION: As an object of "governmental technologies", the nursing staff becomes the body onto which a process of conforming to the customs of the correctional milieu is dictated and inscribed. The results of this qualitative research, from a nursing perspective, are the first of their kind to be reported in Canada since the creation of the Regional Psychiatric Correctional Units in 1978. PMID- 15822848 TI - Do suicide survivors suffer social stigma: a review of the literature. AB - One of the delineating elements found in suicide bereavement versus normal bereavement is the stigma experienced by survivors. This review of the literature will provide insight into stigma as an underlying element in suicide bereavement and point to the role of health professionals in dealing with this complex issue. Historical review and empirical studies are analyzed to provide a framework for how suicide relates to natural bereavement. The conclusion is that suicide bereavement is different from natural loss. The challenge to health care providers is to sort through the complex issues surrounding the individual and their social network to find mechanisms that lead to resolution. Suicide has a profound effect on the family, friends, and associates of the victim that transcends the immediate loss. As those close to the victim suffer through bereavement, a variety of reactions and coping mechanisms are engaged as each individual sorts through individual reactions to the difficult loss. Bereavement refers to "all the physiological, psychological, behavioral, and social response patterns displayed by an individual following the loss (usually through death) of a significant person or thing" (Dunne, Dunne-Maxim & McIntosh, 1987). Bereavement following suicide is complicated by the complex psychological impact of the act on those close to the victim. It is further complicated by the societal perception that the act of suicide is a failure by the victim and the family to deal with some emotional issue and ultimately society affixes blame for the loss on the survivors. This individual or societal stigma introduces a unique stress on the bereavement process that in some cases requires clinical intervention. PMID- 15822849 TI - Mediation of the relationship between inner voice experiences and health-related quality of life. AB - PROBLEM: Appraisals tied to voice hearing and other subjective experiences pose serious challenges for nursing because of their implications for health and safety of voice hearers and others. METHOD: An exploratory, correlational design involving hierarchical analysis of data from 337 voice hearers. FINDINGS: Inner voice experiences and subjective-deficit symptoms each had significant negative, independent effects on perceptions of the integrity of the functional nervous system, self-esteem, and health-related quality of life. The combination of subjective-deficit symptoms, perceptions of the integrity of the functional nervous system and self-esteem significantly reduced the negative impact of inner voice experiences on health related quality of life (R2 = .511). Perceived integrity of the functional nervous system and self-esteem together almost completely attenuated the impact of voice hearing on subjects' health-related quality of life. CONCLUSION: These results highlight the need to develop strategies that can help voice hearers respond to inner experiences in a more positive manner. PMID- 15822850 TI - Uniting the family of psychiatric nurses: commonalities and divergences in the nursing lives we lead. AB - We have created a new column that will focus on the state of psychiatric nursing around the world and on the increasingly important need for all of us, the family of psychiatric nurses, to begin to explore more deeply, and strengthen in more focused ways, the ties that bind all of us. We begin with an article that presents an international perspective on psychiatric illness and mental health promotion and ends by identifying common concerns often shared by the world's psychiatric nurses. Susan McCabe has a unique ability to blend the hard-core facts with a compassionate and intelligent point of view that is compelling and understandable. She is currently an associate professor at the Fay W. Whitney School of Nursing, University of Wyoming. She is an international speaker who presented at many international venues including the Sigma Theta Tau International Nursing Research Conference in the Virgin Islands in 2003 and the International Conference for Mental Health Nurses in Malta in 2002, a gathering that explored and discussed global issues of mental illness and psychiatric nursing. She is also frequently a keynote speaker or presenter at numerous psychiatric nursing conferences and has published extensively in several refereed journals and book chapters. She has expressed a deep desire for other psychiatric nurses to add their voices to hers and to comment and dialogue about our practice lives via the journal. PMID- 15822851 TI - Questions and answers for the nurse psychotherapist in private practice. PMID- 15822853 TI - [Basically, each day is different]. PMID- 15822852 TI - The art of prescribing. Principles of switching novel antipsychotic medications. AB - This case history is one example of how patients can be switched to novel antipsychotic medications in persons with chronic and stable schizophrenia. This brief discussion offers several guidelines for switching novel antipsychotic medications. The decision to switch medications must be based on individual patient needs, variances, and preferences. Clinical decisions must also be guided by current clinical data from psychiatric and physical examinations and treatment goals that improve quality of life and psychosocial functioning. PMID- 15822854 TI - [The whole picture, a link in the chain of care]. PMID- 15822855 TI - [Urgent business for IBODE (the National Union of Nursing Societies)]. PMID- 15822856 TI - Examination of the Lunatic fringe and Uncx4.1 expression by whole-mount in situ hybridization in the embryo of the CKH-Jsr (jumbled spine and ribs) mouse. AB - The CKH-Jsr (jumbled spine and ribs) mouse was found as a spontaneous mutant with malformation of vertebrae, that is, a short trunk and kinky tail. We examined Lunatic Fringe (Lfng) and Uncx4.1 expression in the presomitic mesoderm (PSM) and somites of Jsr-mutant (CKH-Jsr/+) embryos to elucidate pathogenesis of the Jsr mutation. Expression pattern of Lfng in the PSM of Jsr-mutant embryos was similar to that of the normal (C57BL/6) embryos. However, expression pattern of Uncx4.1 in the somites of Jsr-mutant embryos was impaired to be irregular and mosaic, suggesting that the anterior-posterior (A-P) polarity is disordered in the Jsr mutant. These results indicate that the Jsr mutation disrupts the A-P polarity of somites during the somitogenesis without altering Lfng expression pattern in the PSM. PMID- 15822857 TI - Viral and bacterial diseases in livestock in Mongolia. AB - This review focuses on the status of infectious diseases that are serious for animal health and have adverse economic effects in Mongolia. Data presented here are limited due to the lack of published or other easily available documents. Foot-and-mouth disease continues to cause substantial economic losses as exemplified by the outbreak of infection with serotype O PanAsia lineage virus. In the case of the 2001 outbreak, a 65% reduction in export revenues was recorded. In order to ascertain the free status of Mongolia from rinderpest, sero epidemiological surveillance has been carried out since 2001. In 2004, Mongolia was certified free from rinderpest by Office International des Epizooties (OIE). A sharp rise in both animal and human brucellosis incidence has become a serious problem. Rabies and anthrax remain endemic with occasional human cases. Other prevailing infectious diseases are contagious pustular dermatitis, contagious agalactia, enterotoxemia and pasteurellosis. The current programs for the control of infectious diseases in livestock in Mongolia lack a definite policy that would enable rapid implementation. A large-scale surveillance of infectious diseases in animals and management of appropriate preventive measures are urgently required in Mongolia. PMID- 15822858 TI - Antigenotoxic effect of Pleurotus cornucopiae extracts on the mutagenesis of Salmonella typhimurium TA98 elicited by benzo[a]pyrene and oxidative DNA lesions in V79 hamster lung cells. AB - Pleurotus cornucopiae (PC) mushroom with a brilliant yellow pileus is found in the field and known in Japan as Tamogi dake mushroom. The purpose of this paper is to investigate the mechanism of the antimutagenic effect of PC mushroom using both the Ames test and Comet assay. We have found a strong inhibitory effect of both aqueous and organic PC extracts on the mutagenicity elicited by benzo[a]pyrene (B[a]P). This inhibition was dose-dependent in reaction mixtures containing cytosolic and microsomal fractions (S-9) from untreated rat liver as well as in those containing S-9 from aryl hydrocarbon receptor (Ah) ligand of Sudan III-treated rats. Sudan III was a potent inducer of cytochrome P450 1A (CYP1A) activity. We treated rats with Sudan III to enhance the metabolic activation of B[a]P by the S-9 fraction. To explain whether this antimutagenicity was due to the inhibition of CYP1A activity that metabolically activates B[a]P, we tested the effects of the extracts on activities of CYP1A1 and CYP1A2, represented by ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O demethylase (MROD), respectively. Both aqueous and organic extracts inhibited EROD activity at all dose levels, while the inhibitory effect was only observed at high doses with regard to MROD activity. Furthermore, pre-treatment of Chinese hamster V79cells with PC extracts significantly reduced H2O2-induced-DNA damage, indicating that PC extracts provide a protective effect against oxidative DNA damage. These results indicate that whole-mushroom extracts contain compounds that may inhibit the metabolic activation of B[a]P by CYP1A1 as well as prevent oxidative DNA damage. PMID- 15822859 TI - Lycopene and beta-carotene ameliorate catechol estrogen-mediated DNA damage. AB - The consumption of fruits and vegetables is associated with a reduced risk of various ailments, including cancer and cardiovascular diseases. Carotenoids, such as lycopene and beta-carotene, are natural constituents of edible plants and may protect against disease. In this study, the influence of lycopene and beta carotene on DNA damage caused by catechol-estrogens in vitro is examined. One possible mechanism by which catechol estrogens such as 4-hydroxyestradiol (4 OHE2) and 2-hydroxyestradiol, which cause DNA damage in naked plasmid DNA as well as in cells, contributing to the process of carcinogenesis, is through the generation of reactive oxygen species. It was found that both carotenoids at concentrations ranging from 0.25 to 10 microM significantly inhibit strand breakage induced by 4-OHE2/copper sulphate by up to approximately 90% in plasmid DNA with beta-carotene being slightly more effective. No prooxidant or cytotoxic effects were observed at the concentrations tested. These carotenoids had a similar, though reduced effect on DNA damage as measured by the comet assay, in Chinese hamster lung fibroblasts. The results obtained show that both lycopene and beta-carotene, most probably and mainly through their potent antioxidant properties, are able to inhibit catechol-estrogen-mediated DNA damage. PMID- 15822860 TI - Effect of components of green tea extracts, caffeine and catechins on hepatic drug metabolizing enzyme activities and mutagenic transformation of carcinogens. AB - Green tea contains catechins and caffeine as major constituents. Treatment of rats with green tea (2.5% w/v) significantly increased 7-ethoxycoumarin O deethylase (7-ECOD), caffeine N-1 demethylase (CN1D) and UDP glucuronyltransferase (UGT) activities. Treatment with caffeine similarly activated CYP1A2 and related monooxygenases as well as UGT, while treatment with catechins induced UGT activity but not 7-ECOD or CN1D activity. Numbers of benzo[a]pyrene (BP)-induced revertant colonies in an Ames test (mutation assay) with S. typhimurium TA98 as the test strain were markedly larger when BP was preincubated with the liver S-9 (9000 x g supernatant of liver homogenate) from green tea-treated rats than when preincubated with that from control rats. In a modified Ames assay system in which UGT was activated by the addition of UDP glucuronic acid to the preincubation mixture, the numbers of revertant colonies in the assay using liver S-9 from green tea-treated rats decreased to a similar level to that in the assay using S-9 from controls. The acceleration of two enzymatic reactions may contribute to the rapid elimination of BP; the first step, the formation of a metabolic intermediate (which is mutagenic) by CYP1A2 and the second, the conjugation of active metabolic intermediates by UGT. We speculated that green tea can reduce the amount of time carcinogens reside in the body and the chance that body tissues will be exposed to active metabolites of carcinogens thorough rapid elimination due to the simultaneous induction of CYP1A2 and UGT activities. PMID- 15822861 TI - [Andrew Schally, discover of GnRH]. PMID- 15822862 TI - [Andrew Schally and his discoveries]. PMID- 15822863 TI - [Pharmacodynamics of triptorelin]. PMID- 15822864 TI - [Recent research on GnRH receptors]. PMID- 15822865 TI - [GnRH resistance and the gene GPR54]. PMID- 15822866 TI - [From the discovery of hypothalamic hormones to the development of analogues as anti-cancer agents]. PMID- 15822867 TI - [Gonadorelin antagonists: present and future ]. PMID- 15822868 TI - [GnRH antagonists and prostatic hyperplasia]. PMID- 15822869 TI - [GnRH agonists versus antagonists in fertilization in vitro]. PMID- 15822870 TI - [GnRH analogues in the treatment of prostate cancer]. PMID- 15822871 TI - [GnRH analogs and breast cancer]. PMID- 15822872 TI - [Treatment of precocious puberty by GnRH agonists: new methods]. PMID- 15822873 TI - Radiological aspects of leukaemia in childhood: an analysis of 89 children. PMID- 15822874 TI - The dorsum sellae in infancy and childhood. PMID- 15822875 TI - Normal position of the aqueduct of sylvius in infancy. PMID- 15822876 TI - Cranial response to treatment of sagittal synostosis. PMID- 15822877 TI - Opacification of the renal parenchyma in obstruction and reflux. PMID- 15822878 TI - Catheter-related complications of total parenteral nutrition in infants. PMID- 15822879 TI - The dorsal defect of the patella. PMID- 15822880 TI - Bone changes of unknown etiology affecting phalanges of fingers in children: report of eight cases. PMID- 15822881 TI - Neonatal scurvy: report of a case. PMID- 15822882 TI - Arteriovenous malformation of the vertebral artery. PMID- 15822883 TI - Congenital eventration of the septum transversum: a rare form of congenital diaphragmatic anomaly. PMID- 15822884 TI - Sudden blindness as a complication of sphenoid sinusitis: a case report. PMID- 15822885 TI - Development anomalies of the occiput. PMID- 15822886 TI - Azygos vein dilatation in acquired obstruction of the inferior vena cava (examples following calcified caval thrombosis and following mesocaval shunt for portal hypertension). PMID- 15822887 TI - Postoperative radiographic findings following systemic-pulmonary arterial anastomosis in the first week of life. PMID- 15822888 TI - Spontaneous air embolism in the newborn: a complication of pulmonary interstitial emphysema. PMID- 15822889 TI - Congenital pulmonary lymphangiectasis. Report of two cases. PMID- 15822890 TI - Perforation of the gastrointestinal tract and pneumoperitoneum in newborns treated with continuous lung distending pressures. PMID- 15822891 TI - Incarcerated inguinal hernia in infants: plain films and barium enema. PMID- 15822892 TI - Congenital megacalyces. PMID- 15822893 TI - Neonatal congestive heart failure due to mediastinal cyst. PMID- 15822894 TI - Ascites due to spontaneous rupture of the renal pelvis in an 11-month-old infant with uretero-pelvic junction obstruction. PMID- 15822895 TI - Avulsed vertebral rim apophysis in a child. PMID- 15822896 TI - Increased levels of ubiquitin in the 6-OHDA-lesioned striatum of rats. AB - Multiple genetic deficits have linked impaired ubiquitin-conjugation pathways to various forms of familiar Parkinson's disease. We therefore examined the possible role of 6-hydroxydopamine, a dopaminergic neurotoxin used in Parkinson's disease experimental models, in causing protein degradation and its association with the ubiquitin proteasome system. Using unilaterally 6-hydroxydopamine-denervated rats and mass spectrometry profiling directly on brain tissue sections, we here report for the first time an increased level of unconjugated ubiquitin specifically in the dorsal striatum of the dopamine depleted hemisphere. No similar changes were found in the intact hemisphere or in the ventral striatum of the dopamine depleted hemisphere. The lesioning of the dopamine innervation to the striatum was confirmed by a strongly reduced dopamine transporter binding in the striatum, indicating an abundant loss of dopamine neurons. These results suggest that denervation of dopamine neurons per se is implicated in the regulation of ubiquitin pathways, at least in a classical animal model of Parkinson's disease. This study adds additional information regarding the involvement of the ubiquitin proteasome system in Parkinson's disease. PMID- 15822897 TI - Proteomic analysis of serum cytokine levels in response to highly active antiretroviral therapy (HAART). AB - A 30-cytokine protein microarray was used to screen for cytokine profile changes in HIV-infected patients in response to highly active antiretroviral therapy (HAART). Serum cytokines showing significant changes were confirmed by enzyme immunoassay. Monokine induced by gamma-interferon (MIG) and interferon-inducible protein-10 (IP-10) levels significantly decreased after 24 weeks of HAART. Protein microarrays are useful for initial screening of novel cytokine expression. Further studies are needed to elucidate the role of MIG and IP-10 in response to HAART. PMID- 15822898 TI - Effect of biofilm model, mode of growth, and strain on streptococcusmutans protein expression as determined by two-dimensional difference gel electrophoresis. AB - The effect of biofilm model, strain and mode of growth (biofilm or planktonic) on protein expression in Streptococcus mutans, a dental pathogen, was determined by two-dimensional difference gel electrophoresis. The bacterial strain (21-28% differentially expressed proteins) and the biofilm model (0.3-7.8% differential expression) used have a much larger effect on protein expression than the mode of growth (0.2-0.7% differential expression), something that has been ignored in biofilm studies up to now. PMID- 15822899 TI - Analysis of the dorsal spinal cord synaptic architecture by combined proteome analysis and in situ hybridization. AB - The proteomic analysis of tissue samples is an analytical challenge, because identified gene products not only have to be assigned to subcellular structures, but also to cell subpopulations. We here report a strategy of combined subcellular proteomic profiling and in situ hybridization to assign proteins to subcellular sites in subsets of cells within the dorsal region of rat spinal cord. With a focus on synaptic membranes, which represent a complex membrane protein structure composed of multiple integral membrane proteins and networks of accessory structural proteins, we also compared different two-dimensional gel electrophoresis systems for the separation of the proteins. Using MALDI mass spectrometric protein identification based on peptide mass fingerprints, we identified in total 122 different gene products within the different synaptic membrane subfractions. The tissue structure of the dorsal region of the spinal cord is complex, and different layers of neurons can be distinguished neuroanatomically. Proteomic data combined with an in situ hybridization analysis for the detection of mRNA was used to assign selected gene products, namely the optical atrophy protein OPA-1, the presynaptic cytomatrix protein KIAA0378/CAST1, and the uncharacterized coiled-coil-helix-coiled-coil-helix domain containing protein 3 (hypothetical protein FLJ20420), to cell subsets of the dorsal area of the spinal cord. Most striking, KIAA0378/CAST1 mRNA was found only sparsely within the dorsal horn of the spinal cord, but highly abundant within the dorsal root ganglion. This finding, combined with the identification of KIAA0378/CAST1 within the synaptic membrane fraction of the spinal cord at the protein level, are consistent with the reported presynaptic localization of CAST, predominantly within the tissue we investigated primarily attributable to primary afferent sensory neurons. Our approach may be of use in broader studies to characterize the proteomes of neural tissue. PMID- 15822900 TI - Proteome analysis of membrane and cell wall associated proteins from Staphylococcus aureus. AB - Pathogenesis of Staphylococcus aureus, an opportunistic human pathogen, is complex and involves many virulence factors including an array of surface proteins (adhesins) that promote bacterial interactions with extracellular matrix components. A better understanding of these interactions can be achieved by studying the expression of membrane and cell wall associated proteins using a proteome analysis approach. To accomplish this, our goal here was to construct a reference map of membrane and cell wall associated proteins for S. aureus. Various lytic and solubilization methods have been tested to identify a suitable methodology for detection of these proteins in two-dimensional electrophoresis (2DE). Results demonstrate that cell lysis with lysostaphin, which lyses staphylococcal peptidoglycan, followed by solubilization with urea, thiourea, amidosulfobetaine 14 (ASB 14) and dithiothreitol (DTT) is an effective method, yielding a sample comprising proteins of wide molecular ranges and isoelectric points with minimum contamination from cytosolic proteins. Mass spectrometric analysis was employed to identify the membrane and cell surface proteins present in the sample and consequently an initial proteomic map of membrane and cell wall associated proteins for S. aureus is presented. PMID- 15822901 TI - Localization of an N-domain region of angiotensin-converting enzyme involved in the regulation of ectodomain shedding using monoclonal antibodies. AB - ACE chimeric proteins and N domain monoclonal antibodies (mAbs) were used to determine the influence of the N domain, and particular regions thereof, on the rate of ACE ectodomain shedding. Somatic ACE (having both N and C domains) was shed at a rate of 20%/24 h. Deletion of the C domain of somatic ACE generated an N domain construct (ACEDeltaC) which demonstrated the lowest rate of shedding (12%). However, deletion of the N domain of somatic ACE (ACEDeltaN) dramatically increased shedding (212%). Testicular ACE (tACE) having 36 amino acid residues (heavily O-glycosylated) at the N-terminus of the C domain shows a 4-fold decrease in the rate of shedding (49%) compared to that of ACEDeltaN. When the N terminal region of the C domain was replaced with the corresponding homologous 141 amino acids of the N domain (N-delACE) the rate of shedding of the ACEDeltaN was only slightly decreased (174%), but shedding was still 3.5-fold more efficient than wild-type testicular ACE. Monoclonal antibodies specific for distinct, but overlapping, N-domain epitopes altered the rate of ACE shedding. The mAb 3G8 decreased the rate of shedding by 30%, whereas mAbs 9B9 and 3A5 stimulated ACE shedding 2- to 4-fold. Epitope mapping of these mAbs in conjunction with a homology model of ACE N domain structure, localized a region in the N-domain that may play a role in determining the relatively low rate of shedding of somatic ACE from the cell surface. PMID- 15822902 TI - Simple protein complex purification and identification method for high-throughput mapping of protein interaction networks. AB - Most current methods for purification and identification of protein complexes use endogenous expression of affinity-tagged bait, tandem affinity tag purification of protein complexes followed by specific elution of complexes from beads, and gel separation and in-gel digestion prior to mass spectrometric analysis of protein interactors. We propose a single affinity tag in vitro pull-down assay with denaturing elution, trypsin digestion in organic solvent, and LC-ESI MS/MS protein identification using SEQUEST analysis. Our method is simple and easy to scale-up and automate, making it suitable for high-throughput mapping of protein interaction networks and functional proteomics. PMID- 15822903 TI - Genomic messaging system language including command extensions for clinical data categories. AB - This paper, in the area of clinical bioinformatics, highlights relatively efficient means of storing, exchanging, protecting, and searching human and other genomic data, so as to make the data securely accessible to researchers while respecting patient privacy. One important idea is that the GMSL language can be considered as an extension of the way DNA and protein sequences are written so as to carry with them the wishes of the patient in regard to fine-grained consent (as well as retaining the medical experts' cautions, instructions for use, and annotation), and this is carried, whatever environment (e.g., XML) that the data is from or whatever it is going to. At the deepest level, a stream of data expressed in GMSL resembles highly compressed stream of self-checking machine code. For the reader less familiar with the computational aspects, some simple examples illustrate how the raw language looks and works as a raw stream of (interpreted) bytes. The bioinformatics applications are not confined to the clinical domain. This paper completes the initial specification of the language as previously presented and reports on some important extensions including clinical data categories. PMID- 15822904 TI - Assessing data quality of peptide mass spectra obtained by quadrupole ion trap mass spectrometry. AB - An algorithm is introduced to assess spectral quality for peptide CID spectra acquired by a quadrupole ion trap mass spectrometer. The method employs a quadratic discriminant function calibrated with manually classified 'bad' and 'good' quality spectra, producing a single 'spectral quality' score. Many spectra examined that do not have significant matches are assessed to have good spectral quality, indicating that advances in search methods may yield substantial improvements in results. PMID- 15822905 TI - Phosphoproteomic analysis of synaptosomes from human cerebral cortex. AB - Protein phosphorylation is a crucial post-translational modification mechanism in the regulation of synaptic organization and function. Here, we analyzed synaptosome fractions from human cerebral cortex obtained during therapeutic surgery. To minimize changes in the phosphorylation state of proteins, the tissue was homogenized within two minutes of excision. Synaptosomal proteins were digested with trypsin and phosphopeptides were isolated by immobilized metal affinity chromatography and analyzed by liquid chromatography and tandem mass spectrometry. The method allowed the detection of residues on synaptic proteins that were presumably phosphorylated in the intact cell, including synapsin 1, syntaxin 1, and SNIP, PSD-93, NCAM, GABA-B receptor, chaperone molecules, and protein kinases. Some of the residues identified are the same or homologous to sites that had been previously described to be phosphorylated in mammals whereas others appear to be novel sites which, to our knowledge, have not been reported previously. The study shows that new phosphoproteomic strategies can be used to analyze subcellular fractions from small amounts of tissue for the identification of phosphorylated residues for research and potentially for diagnostic purposes. PMID- 15822906 TI - Differential detergent fractionation for non-electrophoretic eukaryote cell proteomics. AB - Differential detergent fractionation (DDF), which relies on detergents to sequentially extract proteins from eukaryotic cells, has been used to increase proteome coverage of 2D-PAGE. Here, we used DDF extraction in conjunction with the nonelectrophoretic proteomics method of liquid chromatography and electrospray ionization tandem mass spectrometry. We demonstrate that DDF can be used with 2D-LC ESI MS2 for comprehensive cellular proteomics, including a large proportion of membrane proteins. Compared to some published methods designed to isolate membrane proteins specifically, DDF extraction yields comprehensive proteomes which include twice as many membrane proteins. Two-thirds of these membrane proteins have more than one trans-membrane domain. Since DDF separates proteins based upon their physicochemistry and subcellular localization, this method also provides data useful for functional genome annotation. As more genome sequences are completed, methods which can aid in functional annotation will become increasingly important. PMID- 15822907 TI - Application of isotope coded affinity tag (ICAT) analysis for the identification of differentially expressed proteins following infection of atlantic salmon (Salmo salar) with infectious hematopoietic necrosis virus (IHNV) or Renibacterium salmoninarum (BKD). AB - Aquaculture and commercial fisheries worldwide suffer from significant economic loss due to diseases of net-pen reared fish. In British Columbia, infectious hematopoietic necrosis (IHN) and bacterial kidney disease (BKD) epidemics occur because there are currently no commercially available drugs or fully licensed vaccines to treat these diseases. With a better understanding of the molecular mechanisms underlying these diseases, this circumstance might be significantly improved. In the present study, we have used a proteomics approach in an effort to identify and quantitate differentially expressed proteins in the liver and kidneys of diseased and healthy Atlantic salmon (Salmo salar). Isotope coded affinity tagging (ICAT), 2D gel electrophoresis, and multidimensional liquid chromatography coupled to tandem mass spectrometry (2D-LC MS/MS) were used to identify hundreds of differentially expressed proteins. While the direct significance of changes in expression levels of many proteins remains to be elucidated, others appear to be more clearly related to the infectious process. Examples of the latter are discussed here and include, a natural killer cell enhancement factor (NKEF), procathepsin L, superoxide-producing NADPH oxidase and interferon-induced viral resistance protein Mx (IFI-Mx). PMID- 15822908 TI - Proteomic characterization of rat liver exposed to 2,3,7,8-tetrachlorobenzo-p dioxin. AB - Dioxins are a class of polyhalogenated aromatic hydrocarbons that induce a wide spectrum of toxic responses in experimental animals. In this study, 2,3,7,8 tetrachlorobenzo-p-dioxin (TCDD) was exposed to two SD rat groups; one group for short-term exposure at a single dose of 1, 10, 20 and 50 mug/kg body weight (group 1) and the other for long-term exposure at daily and-low dose of 0.01, 0.1, 1 and 2.5 microg/kg body weight (group 2) for a month. Two-dimensional electrophoresis (2-DE) was utilized to resolve the protein profile of rat liver exposed to TCDD at different doses. In the analysis of 2-DE of the group 1, two new-expressed spots and seven volume-increased spots were detected and identified by ESI-Q-TOF MS/MS; especially, proteasome subunit beta type 3 was increased in all doses. In addition, in the group 2, six volume-increased spots were screened; particularly, histidine triad nucleotide binding protein was increased in both 0.1 microg/kg dose and 1 microg/kg dose. The identified proteins were confirmed using Western blot. Among the identified proteins, apolipoprotein A-IV may protect lipid peroxidation and atherosclerosis induced by TCDD exposure and the expression level of phosphoglycerate mutase increases due to hyperthyroidism induced by TCDD exposure. PMID- 15822909 TI - Effect of high density lipoproteins on protein expression in myoblast cell lines. AB - We have studied the effects of high-density lipoprotein (HDL) on rat heart (H9c2) and skeletal (L6) myoblasts using two-dimensional gel electrophoresis and tandem mass spectrometry. Gels generated from control cells and cells treated with 250 microg/mL HDL showed significant differences in the 7-10 pI region and the 30-50 kDa mass region. In particular, the membrane binding protein, annexin II, the voltage-dependent anion channel, glyceraldehyde-3-phosphate dehydrogenase and other glycolytic proteins are differentially expressed. PMID- 15822910 TI - Proteomic analysis of lipid microdomains from lipopolysaccharide-activated human endothelial cells. AB - The endothelium plays a critical role in orchestrating the inflammatory response seen during sepsis. Many of the inflammatory effects of Gram-negative sepsis are elicited by lipopolysaccharide (LPS), a glycolipid component of bacterial cell walls. Lipid-rich microdomains have been shown to concentrate components of the LPS signaling system. However, much remains to be learned about which proteins are constituents of lipid microdomains, and how these are regulated following cell activation. Progress in this area would be accelerated by employing global proteomic analyses, but the hydrophobicity of membrane proteins presents an analytical barrier to the effective application of such approaches. Herein, we describe a method to isolate detergent-resistant membranes from endothelial cells, and prepare these samples for proteomic analysis in a way that is compatible with subsequent separations and mass spectrometric (MS) analysis. In the application of these sample preparation and MS analyses, 358 proteins from the lipid-rich microdomains of LPS-activated endothelial cell membranes have been identified of which half are classified as membrane proteins by Gene Ontology. We also demonstrate that the sample preparation method used for solubilization and trypsin digestion of lipid-rich microdomains renders the membrane spanning sequences of transmembrane proteins accessible for endoproteolytic hydrolysis. This analysis sets the analytical foundation for an in-depth probing of LPS signaling in endothelial cells. PMID- 15822911 TI - P-Mod: an algorithm and software to map modifications to peptide sequences using tandem MS data. AB - The discovery of unanticipated protein modifications is one of the most challenging problems in proteomics. Whereas widely used algorithms such as Sequest and Mascot enable mapping of modifications when the mass and amino acid specificity are known, unexpected modifications cannot be identified with these tools. We have developed an algorithm and software called P-Mod, which enables discovery and sequence mapping of modifications to target proteins known to be represented in the analysis or identified by Sequest. P-Mod matches MS/MS spectra to peptide sequences in a search list. For spectra of modified peptides, P-Mod calculates mass differences between search peptide sequences and MS/MS precursors and localizes the mass shift to a sequence position in the peptide. Because modifications are detected as mass shifts, P-Mod does not require the user to guess at masses or sequence locations of modifications. P-Mod uses extreme value statistics to assign p value estimates to sequence-to-spectrum matches. The reported p values are scaled to account for the number of comparisons, so that error rates do not increase with the expanded search lists that result from incorporating potential peptide modifications. Combination of P-Mod searches from multiple LC-MS/MS analyses and multiple samples revealed previously unreported BSA modifications, including a novel decarboxymethylation or D-->G substitution at position 579 of the protein. P-Mod can serve a unique role in the identification of protein modifications both from exogenous and endogenous sources and may be useful for identifying modified protein forms as biomarkers for toxicity and disease processes. PMID- 15822912 TI - Genistein reverses changes of the proteome induced by oxidized-LDL in EA.hy 926 human endothelial cells. AB - Endothelial cells are primary targets for pro-atherosclerotic stressors such as oxidized LDL (ox-LDL). The isoflavone genistein, on the other hand, is suggested to prevent a variety of processes underlying atherosclerosis and cardiovascular diseases. By analyzing the proteome of EA.hy 926 endothelial cells, here we show, that genistein reverses the ox-LDL-induced changes of the steady-state levels of several proteins involved in atherosclerosis. These alterations caused by genistein are functionally linked to the inhibition of ox-LDL induced apoptosis. PMID- 15822913 TI - Search for cancer markers from endometrial tissues using differentially labeled tags iTRAQ and cICAT with multidimensional liquid chromatography and tandem mass spectrometry. AB - A total of nine potential markers for endometrial cancer (EmCa) have been discovered and identified from endometrial tissue homogenates using a combination of differentially labeled tags, iTRAQ and cICAT, with multidimensional liquid chromatography and tandem mass spectrometry. The tissues were snap frozen in liquid nitrogen within 15-20 min after devitalization. Samples for proteomic analysis were treated with protease inhibitors before processing. Marker proteins that were overexpressed in EmCa are chaperonin 10, pyruvate kinase M1 or M2 isozyme, calgizzarin, heterogeneous nuclear ribonucleoprotein D0, macrophage migratory inhibitory factor, and polymeric immunoglobulin receptor precursor; those that were underexpressed are alpha-1-antitrypsin precursor, creatine kinase B, and transgelin. The chaperonin 10 result confirms our earlier observation of overexpression in EmCa tissues using surface-enhanced laser desorption/ionization mass spectrometry, verified by Western analysis and immunohistochemistry [Yang, E. C. C. et al. J. Proteome Res. 2004, 3, 636-643]. Pyruvate kinase was observed to be overexpressed using both iTRAQ and cICAT labeling. All nine markers have been found to be associated with various forms of cancer. A panel of these plus other markers may confer sufficient selectivity for diagnosing and screening of EmCa. The use of cICAT led to identification of a higher proportion of lower abundance signaling proteins; conversely, iTRAQ resulted in a higher percentage of the more abundant ribosomal proteins and transcription factors. PMID- 15822914 TI - Protein identification in complex mixtures. AB - This paper investigates the prospects of successful mass spectrometric protein identification based on mass data from proteolytic digests of complex protein mixtures. Sets of proteolytic peptide masses representing various numbers of digested proteins in a mixture were generated in silico. In each set, different proteins were selected from a protein sequence collection and for each protein the sequence coverage was randomly selected within a particular regime (15-30% or 30-60%). We demonstrate that the Probity algorithm, which is characterized by an optimal tolerance for random interference, employed in an iterative procedure can correctly identify >95% of proteins at a desired significance level in mixtures composed of hundreds of yeast proteins under realistic mass spectrometric experimental constraints. By using a model of the distribution of protein abundance, we demonstrate that the very high efficiency of identification of protein mixtures that can be achieved by appropriate choices of informatics procedures is hampered by limitations of the mass spectrometric dynamic range. The results stress the desire to choose carefully experimental protocols for comprehensive proteome analysis, focusing on truly critical issues such as the dynamic range, which potentially limits the possibilities of identifying low abundance proteins. PMID- 15822915 TI - Reproducibility of tryptic digestion investigated by quantitative fourier transform ion cyclotron resonance mass spectrometry. AB - In this study, the reproducibility of tryptic digestion of complex solutions was investigated using liquid chromatography Fourier transform ion cyclotron resonance (LC FT-ICR) mass spectrometry. Tryptic peptides, from human cerebrospinal fluid, (CSF) were labeled with Quantification-Using-Enhanced-Signal Tags (QUEST)-markers, or 1-([H4]nicotinoyloxy)- and 1-([D4]nicotinoyloxy) succinimide ester markers. The analysis was performed on abundant proteins with respect-to-intensity ratios and sequence coverage and obtained by comparing differently labeled components from one or different pools. To interpret the dynamics in the proteome, one must be able to estimate the error introduced in each experimental steps. The intra sample variation due to derivatization was approximately 10%. The inter sample variation depending on derivatization and tryptic digestion was not more than approximately 30%. These experimental observations provide a range for the up- and down-regulations that are possible to study with electrospray ionization LC FT-ICR mass spectrometry. PMID- 15822916 TI - Proteomic profiling of surface proteins on Th1 and Th2 cells. AB - We utilized mass spectrometry to profile cell surface protein differential expression on primary human T helper (Th1 and Th2) cells with the stable isotope labeling by amino acids in cell culture (SILAC) approach. Proteomic and microarray analyses were done concurrently and results were compared for 38 different genes. Although microarray studies displayed wide variability between donors for mRNA expression, these two approaches were shown to be corroborative for most gene products with the exception of a small subset of uncorrelated protein and message levels. The greatest differing Th1 to Th2 ratios were observed for BST2 (bone marrow stromal protein 2) and TRIM (T cell receptor interacting molecule). Both showed greater Th1 expression by proteomic methods, even though mRNA levels were approximately equal for both. To validate this method, we compared protein expression levels of a recently cloned molecule, B and T cell lymphocyte attenuator (BTLA), on Th1 and Th2 cell populations and showed greater protein expression on Th1 cells, which agrees with a previous analysis of higher BTLA mRNA expression in Th1 cells.(1). PMID- 15822917 TI - Depletion of high-abundant proteins in body fluids prior to liquid chromatography fourier transform ion cyclotron resonance mass spectrometry. AB - Today, proteomics is an exciting approach to discover potential biomarkers of different disorders. One challenge with proteomics experiments is the wide concentration range of proteins in various tissues and body fluids. The most abundant component in human body fluids, human serum albumin (HSA), is present at concentrations corresponding to approximately 50% of the total protein content in, e.g., plasma and cerebrospinal fluid (CSF). If this component could be selectively removed, then the chances of observing lower-abundance component of clinical interest would be greatly improved. There are today several approaches of varying specificity available for depletion. In this study, the properties of two commercially available kits, for the removal of HSA and HSA and immunoglobulin G (IgG), respectively, were compared, and the benefits of using depletion steps prior to on-line LC-FTICR MS were evaluated. Both methods were applied on plasma and CSF. To our knowledge, these are the first results reported for CSF. Also, the combination with electrospray LC-FTICR MS is novel. The proportion of depleted HSA and IgG was estimated using global labeling markers for peptide quantification. Both depletion-methods provided a significant reduction of HSA, and the identification of lower abundant components was clearly facilitated. A higher proportion of HSA was removed using the affinity-based removal kit, and consequently more proteins could be identified using this approach. PMID- 15822918 TI - Fluorescence properties of glutamine-binding protein from Escherichia coli and its complex with glutamine. AB - In this work, the fluorescence of glutamine-binding protein (GlnBP) and its complex with glutamine (GlnBP/Gln) in native and unfolded forms was studied. The experimental data were interpreted on the basis of the results of the analysis of Trp and Tyr microenvironments taking into the account the data for GlnBP mutated forms Trp32Phe(Tyr) and Trp220Phe(Tyr), which have been obtained by Axelsen et al. (Biophys. J. 1991, 60, 650-659). This allowed us to explain the negligible contribution of Tyr residues to the bulk fluorescence of the native protein, the similarity of the fluorescence characteristics of GlnBP and GlnBP/Gln, and the uncommon effect of the excess of the fluorescence intensity at 365 nm (Trp emission) upon excitation at 297 nm respect to the excitation at 280 nm. The last effect is explained by the spectral dependence of the Trp 32 and Trp 220 contributions to the protein absorption. The protein Trp fluorescence dependence on the excitation wavelength must be taken into account for the evaluation of the Tyr residues contribution to the bulk fluorescence of protein, and in principle, it also may be used for the development of an approach for the decomposition of a multicomponent protein fluorescence spectrum. PMID- 15822919 TI - Mapping sites of protein phosphorylation by mass spectrometry utilizing a chemical-enzymatic approach: characterization of products from alpha-S1 casein phosphopeptides. AB - A novel chemical-enzymatic approach was developed to facilitate identification of phosphorylation sites in isolated phosphoproteins. ESI-TOF mass spectrometry was used to characterize products from the chemical-enzymatic cleavage of specific phosphorylation sites in bovine alpha-S1 casein and synthetic phosphopeptides containing substitutions at a single phosphorylation site. Further refinements to this approach for identification of protein phosphorylation sites and its utility for the quantification of phosphopeptides by isotope-dilution mass spectrometry are presented. PMID- 15822920 TI - Domain graph of Arabidopsis proteome by comparative analysis. AB - The domain graph of domains and domain combinations of Arabidopsis thaliana is established based on pfam 14.0 database and analyzed via comparison with 10 eukaryotic, 30 bacterial, and 16 archaeal proteomes. The comparative analysis of the domain graphs provides a useful platform for revealing global insights on the evolution of plant kingdom. More importantly, it is a powerful tool for searching not only the possible new function of both plant-specific and nonspecific domains via specific domain combinations in Arabidopsis thaliana but also the functional role of unknown domains. As an example, we present the functional link between ubiquitin and Myb_DNA-binding domains via Bromodomain as the plant specific evidence for the association between transcription and ubiquitin. We further show that PentatricoPeptide Repeats (PPR) proteins have plant-specific links with a wide variety of domains responsible for RNA binding/metabolism, modulation of protein-protein interactions, ubiquitin-conjugation, cell growth/maintenance, catalysis, and others. This further supports the recently proposed association of PPR proteins with specific RNA transcripts and defined effector proteins. Moreover, the domain graph built from tissue-specific genes is frequently associated with DNA binding domains, suggesting that the differentiation of tissue cell types is contributed mostly by tissue-specific transcriptional process. DOGMA (DOmain Graph via coMparitive analysis for Arabidopsis thaliana) is available on-line with a variety of search tools at http://theory.med.buffalo.edu/DOGMA. The database, which allows user-specified search for plant specific domains and their combinations, will be useful as an additional tool for annotation of the proteins that play specific roles in plants and other organisms. PMID- 15822921 TI - Clinical and pharmacogenomic data mining: 3. Zeta theory as a general tactic for clinical bioinformatics. AB - A new approach, a Zeta Theory of observations, data, and data mining, is being forged from a theory of expected information into an even more cohesive and comprehensive form by the challenge of general genomic, pharmacogenomic, and proteomic data. In this paper, the focus is not on studies using the specific tool FANO (CliniMiner) but on extensions to a new broader theoretical approach, aspects of which can easily be implemented into, or otherwise support, excellent existing methods, such as forms of multivariate analysis and IBM's product Intelligent Miner. The theory should perhaps be distinguished from an existing purely number-theoretic area sometimes also known as Zeta Theory, which focuses on the Riemann Zeta Function and the ways in which it governs the distribution of prime numbers. However, Zeta Theory as used here overlaps heavily with it and actually makes use of these same matters. The distinction is that it enters from a Bayesian information theory and data representation perspective. It could thus be considered an application of the 'mathematician's version'. The application is by no means confined to areas of modern biomedicine, and indeed its generality, even merging into quantum mechanics, is a key feature. Other areas with some similar challenges as modern biology, and which have inspired data mining methods such as IBM's Intelligent Miner, include commerce. But for several reasons discussed, modern molecular biology and medicine seem particularly challenging, and this relates to the often irreducible high dimensionality of the data. This thus remains our main target. PMID- 15822922 TI - A proteomic approach to analyze auxin- and zinc-responsive protein in rice. AB - Auxin and zinc are involved in callus and root formation in rice. However, details of the mechanism underlying this process and functional relation between zinc and auxin are unclear. In this study, proteins induced by auxin and zinc in rice were analyzed by a proteomic approach. Root formation on rice seedlings was promoted by 0.45 microM 2,4-D treatment and was further promoted by addition of 260 microM Zn. Microscopic observation revealed that the number of root primodia formed was significantly increased in 2,4-D- and Zn-treated seedlings than that of the control. A total of seven proteins, as analyzed by 2D-PAGE, were increased, and one protein was decreased by 2,4-D and Zn treatment. Expression of elongation factor-1beta' (EF-1beta') both at transcriptional and translational levels was particular abundant in callus and basal parts of young seedlings, and the accumulation of EF-1beta' was consistent with root formation induced by 2,4-D and Zn. Results indicate that higher level of EF-1beta' expression is necessary for auxin- and zinc-induced root formation in rice. PMID- 15822923 TI - Predicted roles for hypothetical proteins in the low-temperature expressed proteome of the Antarctic archaeon Methanococcoides burtonii. AB - Using liquid chromatography-mass spectrometry, 528 proteins were identified that are expressed during growth at 4 degrees C in the cold adapted archaeon, Methanococcoides burtonii. Of those, 135 were annotated previously as unique or conserved hypothetical proteins. We have performed a comprehensive, integrated analysis of the latter proteins using threading, InterProScan, predicted subcellular localization and visualization of conserved gene context across multiple prokaryotic genomes. Functional information was obtained for 55 proteins, providing new insight into the physiology of M. burtonii. Many of the proteins were predicted to be involved in DNA/RNA binding or modification and cell signaling, suggesting a complex, uncharacterized regulatory network controlling cellular processes during growth at low-temperature. Novel enzymatic functions were predicted for several proteins, including a putative candidate gene for the posttranslational modification of the key methanogenesis enzyme coenzyme M methyl reductase. A bacterial-like CRISPR locus was identified as a strong candidate for archaeal-bacterial lateral gene transfer. Gene context analysis proved a valuable augmentation to the other predictive methods in several cases, by revealing conserved gene associations and annotations in other microbial genomes. Our results underscore the importance of addressing the "hypothetical protein problem" for a complete understanding of cell physiology. PMID- 15822924 TI - Cold adaptation of the Antarctic archaeon, Methanococcoides burtonii assessed by proteomics using ICAT. AB - Using isotope coded affinity tag (ICAT) chromatography and liquid chromatography mass spectrometry, 163 proteins were identified from the cold-adapted archaeon, Methanococcoides burtonii. 14 proteins were differentially expressed during growth at 4 degrees C and 23 degrees C. Knowledge of protein abundance, protein identity and gene arrangement was used to determine mechanisms of cold adaptation. Growth temperature was found to affect proteins involved in energy generation and biosynthesis linked to methanogenesis, membrane transport, transcription and protein folding, as well as affecting the expression of two hypothetical proteins. Pooling the data from this ICAT study with data from a previous two-dimensional gel electrophoresis study highlighted consistencies and differences between the two methods, and led us to conclude that the two approaches were generally complementary. This is the first report of ICAT applied to Archaea, or for the study of cold adaptation in any organism. PMID- 15822925 TI - Trypsin-based monolithic bioreactor coupled on-line with LC/MS/MS system for protein digestion and variant identification in standard solutions and serum samples. AB - The applicability of a trypsin-based monolithic bioreactor coupled on-line with LC/MS/MS for rapid proteolytic digestion and protein identification is here described. Dilute samples are passed through the bioreactor for generation of proteolytic fragments in less than 10 min. After digestion and peptide separation, electrospray ionization tandem mass spectrometry is used to generate a peptide map and to identify proteolytic peptides by correlating their fragmentation spectra with amino acid sequences from a protein database. By digesting picomoles of proteins sufficient data from ESI and MS/MS were obtained to unambiguously identify proteins alone and in serum samples. This approach was also extended to locate mutation sites in beta-lactoglobulin A and B variants. PMID- 15822926 TI - SCAPE: a new tool for the Selective CApture of PEptides in protein identification. AB - A method for the selective capture and analysis of peptides containing neither histidine nor arginine is evaluated. It is based on the reversible modification of alpha- and epsilon-amino groups of peptides and the relatively easy separation of charged and noncharged peptides by cation exchange chromatography. The simplicity of the method and the results obtained in silico and with standard proteins, anticipate the utility of the SCAPE approach for proteome analyses. PMID- 15822927 TI - Proteomic characterization of a triton-insoluble fraction from chloroplasts defines a novel group of proteins associated with macromolecular structures. AB - Proteomic analysis of a Triton X-100 insoluble, 30,000 x g pellet from purified pea chloroplasts resulted in the identification of 179 nonredundant proteins. This chloroplast fraction was mostly depleted of chloroplast membranes since only 23% and 9% of the identified proteins were also observed in envelope and thylakoid membranes, respectively. One of the most abundant proteins in this fraction was sulfite reductase, a dual function protein previously shown to act as a plastid DNA condensing protein. Approximately 35 other proteins known (or predicted) to be associated with high-density protein-nucleic acid particles (nucleoids) were also identified including a family of DNA gyrases, as well as proteins involved in plastid transcription and translation. Although nucleoids appeared to be the predominant component of 30k x g Triton-insoluble chloroplast preparations, multi-enzyme protein complexes were also present including each subunit to the pyruvate dehydrogenase and acetyl-CoA carboxylase multi-enzyme complexes, as well as a proposed assembly of the first three enzymes of the Calvin cycle. Approximately 18% of the proteins identified were annonated as unknown or hypothetical proteins and another 20% contained "putative" or "like" in the identifier tag. This is the first proteomic characterization of a membrane depleted, high-density fraction from plastids and demonstrates the utility of this simple procedure to isolate intact macromolecular structures from purified organelles for analysis of protein-protein and protein-nucleic acid interactions. PMID- 15822928 TI - Proteolytic 18O labeling by peptidyl-Lys metalloendopeptidase for comparative proteomics. AB - The potential capabilities of a new proteolytic 18O labeling method employing peptidyl-Lys metalloendopeptidase (Lys-N) have been demonstrated for use in comparative proteomics. Conditions (pH>or=9.5) have been found such that Lys-N incorporates only a single 18O atom into the carboxyl terminus of each proteolytically generated peptide. This 18O labeling method has a major advantage over current protelytic 18O labeling methods that generate a mixture of isotopic isoforms resulting from the incorporation of one or two 18O atoms into each peptide species by the proteases (trypsin, Lys-C, or Glu-C) used. We demonstrate that the single 18O atom incorporation property of Lys-N overcomes the major problem of the current proteolytic 18O labeling methods and provides accurate quantification results for isotopically labeled peptides. PMID- 15822929 TI - Nanoliter sample handling combined with microspot MALDI-MS for detection of gel separated phosphoproteins. AB - We describe a microspot matrix-assisted laser desorption ionization (MALDI) mass spectrometric approach to analyze gel-separated phosphoproteins. This method involves in-gel digestion of phosphoproteins after gel separation, followed by open tubular capillary (OTC) immobilized metal-ion affinity chromatography (IMAC) to capture the phosphopeptides with markedly reduced interferences from nonphosphorylated peptides. Nanoliter-volume of ammonium phosphate is used to elute the phosphopeptides captured on the capillary tube. After mixing with a small volume of matrix solution in the capillary, the effluent is deposited in a microspot on a sample plate for MALDI-MS analysis. It is also shown that, with peptide esterification after in-gel digestion of a phosphoprotein, negative ion detection in MALDI gives a distinct advantage over the positive ion mode of operation for phosphopeptide analysis, even without IMAC enrichment. However, the OTC-IMAC technique is demonstrated to be superior to the approach of negative ion detection of esterified in-gel digests without IMAC. OTC-IMAC is found to be sufficiently selective to capture phosphopeptides from in-gel digest of a gel band containing predominately one protein and the combination of peptide esterification and IMAC enrichment does not provide any real advantage. Using a standard phosphoprotein alpha-casein as a model system, we demonstrate that this OTC-IMAC method can detect a number of phosphopeptides after in-gel digestion with mid-fmol protein sample loading. An example of real world applications of this method is illustrated in the characterization of a fusion protein, His182, expressed in E. coli. PMID- 15822931 TI - Study of the interaction of chlorisondamine and chlorisondamine analogues with an epitope of the alpha-2 neuronal acetylcholine nicotinic receptor subunit. AB - Chlorisondamine (CHL), a neuronal nicotinic ganglionic blocker, when injected in the cerebral ventricle of rats chronically blocks the increase in locomotion and rearing by subcutaneous nicotine injection. The blocking of the ion channel(s) prevents nicotine from exerting its rewarding effects on the CNS. When administered intraperitoneally, a dose 400-500 times the intracerebroventricular one is needed to cross the blood-brain barrier and to generate the same level of nicotine antagonism, resulting in severe side-effects, thus making it unlikely to be used as a therapeutical compound. Three CHL analogues, 2-(indolin-1-yl)-N,N,N trimethylethanaminium iodide, 2-(1,3-dioxoisoindolin-2-yl)- N,N,N trimethylethanaminium iodide, and 2-(1H-indole-3-carboxamido)- N,N,N trimethylethanaminium iodide, were synthesized in the hope of circumventing the parent compound's shortcomings. They all share a modified indole ring, lack the four chlorines CHL carries, and have one tertiary amine and one quaternary amine. The CHL analogues form noncovalent complexes with an epitope of the alpha-2 nicotinic receptor subunit, GEREE(p)TEEEEEEEDEN, previously proposed as the possible site of CHL interaction. Complexes were analyzed using matrix-assisted laser desorption/ionization mass spectrometry for comparison with CHL. Overall, all three analogues showed better affinity than CHL for complex formation with both the nonphosphorylated and phosphorylated epitopes. PMID- 15822930 TI - Quantitative profiling of the detergent-resistant membrane proteome of iota-b toxin induced vero cells. AB - Enzyme-mediated 18O/16O differential labeling of proteome samples often suffers from incomplete exchange of the carboxy-terminus oxygen atoms, resulting in ambiguity in the measurable abundance differences. In this study, an 18O/16O labeling strategy was optimized for and applied to the solution-based comparative analysis of the detergent-resistant membrane proteome (DRMP) of untreated and Iota-b (Ib)-induced Vero cells. Solubilization and tryptic digestion of the DRMP was conducted in a buffer containing 60% methanol. Unfortunately, the activity of trypsin is attenuated at this methanol concentration hampering the ability to obtain complete oxygen atom turnover. Therefore, the incorporation of the 18O atoms was decoupled from the protein digestion step by carrying out the trypsin mediated heavy atom incorporation in a buffer containing 20% methanol; a concentration at which trypsin activity is enhanced compared to purely aqueous conditions. After isotopic labeling, the samples were combined, fractionated by strong cation exchange and analyzed by microcapillary reversed-phase liquid chromatography coupled on-line with electrospray ionization tandem mass spectrometry. In total, over 1400 unique peptides, corresponding to almost 600 proteins, were identified and quantitated, including all known caveolar and lipid raft marker proteins. The quantitative profiling of Ib-induced DRMP from Vero cells revealed several proteins with altered expression levels suggesting their possible role in Ib binding/uptake. PMID- 15822932 TI - Matrix implanted laser desorption ionization (MILDI) combined with ion mobility mass spectrometry for bio-surface analysis. AB - The implantation of low velocity massive gold cluster ions allows homogeneous incorporation of a metallic matrix into the near-surface region of rat brain tissues. Subsequent analysis by laser desorption ionization mass spectrometry yields spectra exhibiting molecular ion peaks in the mass range up to 35 kDa similar to those observed by matrix-assisted LDI. Matrix-implanted LDI when combined with ion-mobility preseparation promises to be a useful technique for molecular imaging of biotissues with a laser microprobe. PMID- 15822933 TI - Identification of protein modifications using MS/MS de novo sequencing and the OpenSea alignment algorithm. AB - Algorithms that can robustly identify post-translational protein modifications from mass spectrometry data are needed for data-mining and furthering biological interpretations. In this study, we determined that a mass-based alignment algorithm (OpenSea) for de novo sequencing results could identify post translationally modified peptides in a high-throughput environment. A complex digest of proteins from human cataractous lens, a tissue containing a high abundance of modified proteins, was analyzed using two-dimensional liquid chromatography, and data was collected on both high and low mass accuracy instruments. The data were analyzed using automated de novo sequencing followed by OpenSea mass-based sequence alignment. A total of 80 modifications were detected, 36 of which were previously unreported in the lens. This demonstrates the potential to identify large numbers of known and previously unknown protein modifications in a given tissue using automated data processing algorithms such as OpenSea. PMID- 15822934 TI - Prediction of peptide retention at different HPLC conditions from multiple linear regression models. AB - To quantitatively characterize the structure of a peptide and to predict its gradient retention time at given HPLC conditions three structural descriptors are used: (i) logarithm of the sum of retention times of the amino acids composing the peptide, log SumAA, (ii) logarithm of the van der Waals volume of the peptide, log VDW(Vol), (iii) and the logarithm of the peptide's calculated n octanol-water partition coefficient, clog P. The log SumAA descriptor is obtained from empirical data for 20 natural amino acids, determined in a given HPLC system. The two other descriptors are calculated from the peptides' structural formulas using molecular modeling methods. The quantitative structure-retention relationships (QSRR), build by multiple linear regression, describe HPLC retention of peptide on a given chromatographic system on which the retention of the 20 amino acids was predetermined. A structurally diversified series of 98 peptides was employed. The predicted gradient retention times on several chromatographic systems were in good agreement with the experimental data. The QSRR equations, derived for a given system operated at variable gradient times and temperatures allowed for the prediction of peptide retention in that system. Matching the experimental HPLC retention to the theoretically predicted for a presumed peptide could facilitate original protein identification in proteomics. In conjunction with MS data, prediction of the retention time for a given peptide might be used to improve the confidence of peptide identifications and to increase the number of correctly identified peptides. PMID- 15822935 TI - Conversion of human alpha-lactalbumin to an apo-like state in the complexes with basic poly-amino acids: toward understanding of the molecular mechanism of antitumor action of HAMLET. AB - It was recently shown that alpha-lactalbumin associated with oleic acid (HAMLET) interacts with core histones thereby triggering apoptosis of tumor cells (J. Biol. Chem. 2003, 278, 42131). In previous work, we revealed that monomeric alpha lactalbumin in the absence of fatty acids can also interact with histones and, moreover, with basic poly-amino acids (poly-Lys and poly-Arg) that represent simple models of histone proteins (Biochemistry 2004, 43, 5575). Association of alpha-lactalbumin with histone or poly-Lys(Arg) essentially changes its properties. In the present work, the character of the changes in structural properties and conformational stability of alpha-lactalbumin in the complex with poly-Lys(Arg) has been studied in detail by steady-state fluorescence, circular dichroism, and differential scanning calorimetry. Complex formation strongly depends on ionic strength, confirming its electrostatic nature. Experiments with the poly-amino acids of various molecular masses demonstrated a direct proportionality between the number of alpha-lactalbumin molecules bound per poly Lys(Arg) and the surface area of the poly-amino acid random coil. The binding of the poly-amino acids to Ca2+-saturated human alpha-lactalbumin decreases its thermal stability down to the level of its free apo-form and decreases Ca2+ affinity by 4 orders of magnitude. The conformational state of alpha-lactalbumin in a complex with poly-Lys(Arg), named alpha-LActalbumin Modified by Poly-Amino acid (LAMPA), differs from all other alpha-lactalbumin states characterized to date, representing an apo-like (molten globule-like) state with substantially decreased affinity for calcium ion. The requirement for efficient conversion of alpha-lactalbumin to the LAMPA state is a poly-Lys(Arg) chain consisting of several tens of amino acid residues. PMID- 15822936 TI - Quantitative proteomic analysis of the secretory proteins from rat adipose cells using a 2D liquid chromatography-MS/MS approach. AB - We have developed two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) and 18O proteolytic labeling strategies to identify and compare levels of secretory proteins with low abundance in the conditioned medium of rat adipose cells without or with insulin stimulation. Culture medium was concentrated and secreted proteins were separated on a RP-HPLC followed by LC MS/MS analysis. For 18O proteolytic labeling, 16O- to 18O-exchange in the digested peptides from eight individual fractions was carried out in parallel in H2(16)O and H(2)18O with immobilized trypsin, and the ratios of isotopically distinct peptides were measured by mass spectrometry. A total of 84 proteins was identified as secreted adipokines. This large number of secretory proteins comprise multiple functional categories. Comparative proteomics of 18O proteolytic labeling allows the detection of different levels of many secreted proteins as exemplified here by the difference between basal and insulin treatment of adipose cells. Taken together, our proteomic approach is able to identify and quantify the comprehensive secretory proteome of adipose cells. Thus, our data support the endocrine role of adipose cells in pathophysiological states through the secretion of signaling molecules. PMID- 15822937 TI - Novel approach for peptide quantitation and sequencing based on 15N and 13C metabolic labeling. AB - Here we describe a method for protein identification and quantification using stable isotopes via in vivo metabolic labeling of the hyperthermophilic crenarchaeon Sulfolobus solfataricus. Stable isotope labeling for quantitative proteomics is becoming increasingly popular; however, its usefulness in protein identification has not been fully exploited. We use both 15N and 13C labeling to create three different versions of the same peptide, corresponding to the unlabeled, 15N and 13C labeled versions. The peptide then appears as three different peaks in a TOF-MS scan and three corresponding sets of MS/MS spectra are obtained. With this information, the elemental carbon and nitrogen compositions for each peptide and each fragment can be calculated. When this is used as a constraint in database searching and/or de novo sequencing, the confidence of a match is increased (for an example intact peptide from 34 choices to 1). This makes the method a useful proteomic tool for both sequenced and unsequenced organisms. Furthermore, it allows for accurate protein quantitation (standard deviations over >4 peptides per protein were within 10%) of three phenotypes in one MS experiment. Abundances for each peptide are calculated by determining the relative areas of each of the three peaks in the TOF-MS spectrum. PMID- 15822938 TI - Identification of the thrombin light chain a as the single best mass for differentiation of gastric cancer patients from individuals with dyspepsia by proteome analysis. AB - Gastric cancer mortality is second only to lung cancer, and its prognosis is dismal. Using surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry, we previously identified a single best mass, which could separate gastric cancer from patients without cancer, with a sensitivity of 89.9% and a specificity of 90%. Using protein liquid chromatography systems with various chromatography media and MS/MS analysis, we were able to identify thrombin light chain A, a proteolytic fragment of prothrombin, as the single best mass for early detection of gastric cancer patients. These findings indicate that disturbances in the coagulation-system are early events in gastric cancer biology and that a decrease or loss of thrombin light chain A, which we termed negative serum protein profiling, may contribute to the diagnosis of cancer patients. PMID- 15822939 TI - High resolution "ultra performance" liquid chromatography coupled to oa-TOF mass spectrometry as a tool for differential metabolic pathway profiling in functional genomic studies. AB - The combination of a new 1.7 mum reversed-phase packing material, and a chromatographic system, operating at ca. 12,000 psi, (so-called ultra performance liquid chromatography, UPLC) has enabled dramatic increases in chromatographic performance to be obtained for complex mixture separation. This increase in performance is manifested in improved peak resolution, together with increased speed and sensitivity. Here, we show that UPLC offers significant advantages over conventional reversed-phase HPLC amounting to a more than doubling of peak capacity, an almost 10-fold increase in speed and a 3- to 5-fold increase in sensitivity compared to that generated with a conventional 3.5 microm stationary phase. The first functional genomic application of UPLC-MS technology is illustrated here with respect to multivariate metabolic profiling of urines from males and females of two groups of phenotypically normal mouse strains (C57BL19J and Alpk:ApfCD) and a "nude mouse" strain. We have also compared this technology to conventional HPLC-MS under similar analytical conditions and show improved phenotypic classification capability of UPLC-MS analysis together with increased ability to probe differential pathway activities between strains as a result of improved analytical sensitivity and resolution. PMID- 15822940 TI - Proteome analysis of antiproliferative mechanism of 12-O-tetradecanoylphorbol 13 acetate on cultured nasopharyngeal carcinoma CNE2 cells. AB - 12-O-Tetradecanoyl-phorbol-13-acetate (TPA) is a plant derivative with multiple function as tumor promoter, differentiation revulsant or leukemia therapy drug. The molecular mechanism of its function is perplexing. Many studies have focused on the mechanism of TPA stimulation in tumor promotion of mouse models or terminal differentiation of leukemia cells, but the effect of TPA on nasopharyngeal carcinoma (NPC) remains unclear, while TPA was considered to be associated with NPC development. In the present study, we employed proteomics techniques to study protein changes of a poorly differentiated squamous carcinoma cell line-CNE2 of human NPCs cells induced by TPA. Six significantly and reproducibly changed proteins were identified and their functional implications were discussed in some details. PMID- 15822941 TI - Eliminating the interferences from TRIS buffer and SDS in protein analysis by fused-droplet electrospray ionization mass spectrometry. AB - Multiply charged protein ions were detected from the solutions containing a high concentration of tris(hydroxymethyl) aminomethane buffer (TRIS) and sodium dodecyl sulfate (SDS) using fused-droplet electrospray ionization mass spectrometry (FD-ESI/MS). The sample aerosols were generated at ambient temperature with a pneumatic nebulizer commonly used to produce sample aerosols in an atmospheric pressure chemical ionization (APCI) source. The aerosols were carried by nitrogen gas to the tip of a capillary where charged methanol droplets had been continuously generated by electrospraying an acidic methanol solution. The neutral sample aerosols then fused with the charged methanol droplets and electrospray ionization proceeded from the newly formed fused droplets to generate multiply charged protein ions. Because of its low solubility in methanol, TRIS molecules (concentration as high as 1 M) were efficiently excluded from the newly formed droplets and the protein ion signals were detected and observed in the mass spectra. To remove the interferences from SDS, equal moles of positively charged cetyltrimethylammonium bromide (CTAB) was added into the SDS containing sample solution to form the dodecyl sulfate-cetyltrimethylammonium ion pair (DS-CTA). The DS-CTA ion pair has a low polarity and solubility in methanol and is excluded from the fused droplet. Protein ions were still detected from the solution containing 10(-2) M of SDS. PMID- 15822942 TI - Human plasma proteome analysis by multidimensional chromatography prefractionation and linear ion trap mass spectrometry identification. AB - A resurgence of interest in the human plasma proteome has occurred in recent years because it holds great promise of revolution in disease diagnosis and therapeutic monitoring. As one of the most powerful separation techniques, multidimensional liquid chromatography has attracted extensive attention, but most published works have focused on the fractionation of tryptic peptides. In this study, proteins from human plasma were prefractionated by online sequential strong cation exchange chromatography and reversed-phase chromatography. The resulting 30 samples were individually digested by trypsin, and analyzed by capillary reversed-phase liquid chromatography coupled with linear ion trap mass spectrometry. After meeting stringent criteria, a total of 1292 distinct proteins were successfully identified in our work, among which, some proteins known to be present in serum in <10 ng/mL were detected. Compared with other works in published literatures, this analysis offered a more full-scale list of the plasma proteome. Considering our strategy allows high throughput of protein identification in serum, the prefractionation of proteins before MS analysis is a simple and effective method to facilitate human plasma proteome research. PMID- 15822943 TI - Metabolic labeling of human primary retinal pigment epithelial cells for accurate comparative proteomics. AB - Metabolic labeling was evaluated, using both 13C6-Arg and 13C6, 15N2-Lys amino acids, for a primary human retinal pigment epithelial cell (hRPE) culture prepared from an autopsy eye of an 81 year old donor. Satisfactory incorporation (>90%) was achieved with both stable isotope labeled amino acids after four passages (roughly 7 population doublings). The degree of incorporation was found to be efficient with both amino acids as well as in different proteins. The presence of 10% whole serum in the culture medium did not interfere with the incorporation of the exogenous stable isotope labeled amino acids. Metabolic labeling of these human primary retinal pigment epithelial cells was further tested to quantify protein ratios between proliferating and resting cells using a combination of 2-DG and MALDI-TOF-TOF/MS analysis. Using computational data processing and analysis, we obtained accurate protein ratio measurement for every single identified protein (156 proteins) in the 2-Dg array. Of these 156 proteins, 12 proteins were found significantly increased in dividing versus resting cells by at least a factor of 1.5 while 13 other proteins were found increased in resting versus dividing cells by at least the same fold. Most of these differentially expressed proteins are directly involved in cell proliferation, protein synthesis, and actin-remodeling and differentiation. PMID- 15822944 TI - Complementary structural information from a tryptic N-linked glycopeptide via electron transfer ion/ion reactions and collision-induced dissociation. AB - Glycosylation is an important post-translational modification. Analysis of glycopeptides is difficult using collision-induced dissociation, as it typically yields only information about the glycan structure, without any peptide sequence information. We demonstrate here how a 3D-quadrupole ion trap, using the complementary techniques of collision induced dissociation (CID) and electron transfer dissociation (ETD), can be used to elucidate the glycan structure and peptide sequence of the N-glycosylated peptide from a fractionated tryptic digest of the lectin from the coral tree, Erythina cristagalli. CID experiments on the multiply protonated glycopeptide ions yield, almost exclusively, cleavage at glycosidic bonds, with little peptide backbone fragmentation. ETD reactions of the triply charged glycopeptide cations with either sulfur dioxide or nitrobenzene anions yield cleavage of the peptide backbone with no loss of the glycan structure. These results show that a 3D-quadrupole ion trap can be used to provide glycopeptide amino acid sequence information as well as information about the glycan structure. PMID- 15822945 TI - Primordial proteins and HIV. AB - Primordial proteins regulate the response of nanobacteria to variations in their environment and reinforce existing pathogenic potentials. By analyzing specific response patterns, we predicted the prevalence of nanobacteria in HIV--and in the atmosphere. A current clinical study indicates the identification of a possibly giant nanobacterial reservoir in Africa: a significant fraction of a test group (40 HIV-infected mothers and 13 babies) was infected with nanobacteria. Concurrently, a multitude of 80-300 nm nanovesicles, apparently nanobacteria, were detected in the atmosphere of the Earth. Nanobacterial infections in HIV are possibly comparable to the twin epidemics HIV and tuberculosis. Models inspired by proteomics recommend methods to inactivate nanobacteria (and other slime producing bacteria) in the body. PMID- 15822946 TI - Analysis of shotgun proteomics and RNA profiling data from Arabidopsis thaliana chloroplasts. AB - The integration of data from transcriptional profiling and shotgun proteomics experiments provides additional information about the identified proteins that goes beyond their plain detection. We have analyzed results from MS/MS shotgun detection of 426 Arabidopsis chloroplast proteins and genome-wide RNA profiling to identify correlations between gene expression, protein abundance and protein characteristics that influence their detection in high-throughput proteome analyses. The integrated data analysis revealed a significant molecular mass bias for the detection of proteins that were expressed at low transcript levels. Overall, the sequence coverage of most of the identified proteins increases with transcript levels indicating a positive correlation between transcript and relative protein abundance. This does not apply to a subset of the identified proteins suggesting specific properties that alter their detection in shotgun proteomics. This integrative comparison is a suitable strategy to validate large scale proteomics data and offers an assessment of the depth of the proteome analysis and the confidence in protein identification. PMID- 15822947 TI - Organohypervalent iodine: development, applications, and future directions. AB - The synthetic utility of organohypervalent iodine reagents will be illustrated by their use in the alpha-hydroxydimethylacetal formation reaction from enolizable ketones, alpha-hydroxylation, alpha-tosyloxylation, alpha-alkoxylation and arylation of ketones, carbon-carbon bond formation, and intramolecular cyclopropanation using iodonium ylides. The uses of these reagents in the Hunsdiecker reaction of carboxylic acids and Hofmann rearrangement of carboxamides is presented. Specific transformation in the cubane series are discussed. The syntheses of a wide range of heterocycle structures are also presented. A unifying pathway for virtually all these diverse reactions is offered; the central features being initial attack at the iodonium center, ligand coupling, with reductive elimination of iodobenzene to yield the product. PMID- 15822948 TI - Transition state studies on the dioxirane-mediated asymmetric epoxidation via kinetic resolution and desymmetrization. AB - This paper describes an efficient kinetic resolution process of trisubstituted cyclic olefins via a chiral dioxirane generated in situ from a fructose-derived ketone and Oxone. The substrates presented include a variety of 1,3-disubstituted and 1,6-disubstituted cyclohexenes with the stereogenic centers at allylic positions. A sequential desymmetrization and kinetic resolution of 1,4 cyclohexadienes by the chiral dioxirane was also found to be feasible. The initially formed monoepoxides can be kinetically resolved by continuing the reaction, leading to the increase or decrease of the ee values of monoepoxides depending on the diene systems. In some cases, a prochiral directing group is not required and the first formed epoxide stereoselectively directs the second epoxidation. When the coupled desymmetrization and kinetic resolution is used synergistically, high enantiopurity can be obtained for an epoxide from an intrinsically less enantioselective substrate. The observed absolute and relative stereochemistry as well as the changing optical purity can be effectively rationalized by transition state analysis. PMID- 15822949 TI - Upper rim allyl- and arylazo-coupled calix[4]arenes as highly sensitive chromogenic sensors for Hg2+ ion. AB - The syntheses and chromogenic properties of calix[4]arenes, carrying 5,17 bisallyl-11,23-bis(p-X-phenyl)azo 3a-c, 5,11,17-triallyl-23-(p-X-phenyl)azo 4a-c, and 5,17-bis(hydroxypropyl)-11,23-bis(p-X-phenyl)azo groups on the upper rims 5a,b, are described. Unexpectedly, UV/vis spectra of the very popular 4-(4 nitrophenyl)azophenol-coupled calix[4]arenes 3c and 4c did not show any shift in lambda(max) when 10 different metal perchlorates were added separately to the host in a methanol-chloroform (v/v = 1/399) cosolvent. In contrast, the absorption spectra of calix[4]arenes with either 4-methoxyphenylazo (3b-5b) or 4 phenylazo (3a-5a) on the upper rim showed substantial bathochromic shifts (Deltalambda = 128-162 nm) upon the addition of soft metal ions (such as Hg(2+), Cr(3+), and Cu(2+)). The 4-(4-methoxyphenyl)azophenol-coupled calix[4]arenes (the 3b-5b series) are found to be highly sensitive for mercury ion (Hg(2+)) among the 10 different metal ions screened. Strong interactions between Hg(2+) ion and the 4-(4-methoxyphenyl)azophenol(s) as well as the p-allyl groups are corroborated by the (1)H NMR studies of 3a,b.Hg(2+) complexes. Furthermore, Job's plots revealed 1:1 binding stoichiometry for all these p-allyl- and arylazo-coupled calix[4]arenes with transition metal ions, and Benesi-Hilderbrand plots were used for the determination of their association constants. PMID- 15822950 TI - 2-(aminomethyl)-oxazolines: highly modular scaffolds for the preparation of novel asymmetric ligands. AB - Highly modular chiral 2-(aminoalkyl)oxazolines have been prepared from alpha amino acids and 1,2-amino alcohols. The amine-functionalized oxazolines were employed as scaffolds in the preparation of a number of different ligands with potential denticities varying from 2 to 5. The obtained ligands were employed and evaluated in the ruthenium-catalyzed asymmetric transfer-hydrogenation of acetophenone and in the titanium-catalyzed addition of diethylzinc to aldehydes. In the latter process, enantioselectivity up to 97% was obtained. PMID- 15822951 TI - Synthesis and stereodynamics of highly constrained 1,8-bis(2,2'-dialkyl-4,4' diquinolyl)naphthalenes (2). AB - Axially chiral 1,8-bis(2,2'-diphenyl-4,4'-diquinolyl)naphthalene, 8, and 1,8 bis(2,2'-diisopropyl-4,4'diquinolyl)naphthalene N,N'-dioxide, 9, have been prepared to study the stereodynamics of these and other 1,8 diheteroarylnaphthalenes based on reversible first-order isomerization kinetics and crystallographic data. The ratio of the two enantiomeric anti-conformers to the meso syn-isomer of 8 and 9 was determined as 1.2:1 and 9.6:1. Investigation of the conformational stability of the atropisomers at enhanced temperatures using HPLC and NMR spectroscopy revealed a Gibbs activation energy of 122.4 (121.8) kJ/mol and 115.2 (109.0) kJ/mol for the anti/syn- (syn/anti) isomerization of 8 and 9, respectively. Comparison of the conformational stability of a series of 1,8-dipyridylnaphthalenes and 1,8-diquinolylnaphthalenes shows that the latter exhibit a significantly higher rotational energy barrier. While the syn- and anti-isomers of 1,8-dipyridylnaphthalenes interconvert rapidly at room temperature the stereoisomers of 1,8-diquinolylnaphthalenes can be isolated by chromatography or crystallization and stored at 25 degrees C for several months without any sign of racemization. The conformational stability of 1,8-diquinolylnaphthalenes is a consequence of significantly increased steric hindrance to isomerization in a highly congested T-shaped transition state. Conversion of 1,8-diheteroarylnaphthalenes to their corresponding N,N'-dioxides was found to result in an increased anti/syn-ratio and decreased rotational energy barrier, which was attributed to synergistic repulsive dipole/dipole interactions destabilizing the diastereomeric ground states and facilitated out of-plane bending reducing the steric hindrance in the T-shaped transition state. PMID- 15822952 TI - Solid-phase methods for the synthesis of cyanine dyes. AB - We report here a series of studies that explore solid-phase methodologies for the synthesis of various cyanine dyes. The scope of the previously reported catch-and release method using sulfonyl chloride resin(1) has now been extended to include pentamethine and water-soluble cyanine dyes. We also report a new and chemically distinct synthetic strategy, employing the stepwise attack of heterocyclic carbon nucleophiles on immobilized polyene-chain precursors, allowing the clean synthesis of hydrophobic and hydrophilic trimethine and pentamethine dyes from more easily obtained starting materials. Overall, both approaches appear to be robust and versatile strategies to delivering a wide range of cyanine-based dyes in high purity. PMID- 15822953 TI - Systematic evaluation of the conformational properties of aliphatic omega-hydroxy acids. AB - A systematic conformational study of omega-hydroxy acids, HO-(CH(2))(n)()-COOH with n ranging from 2 to 5, has been performed using quantum mechanical calculations at the MP2 level. To distinguish between intrinsic and environmentally induced conformational preferences, calculations have been carried out in both gas phase and chloroform solution, a polarizable continuum solvation model being used to represent the latter. Results indicate a consistent conformational behavior, as reflects the feature that the lowest energy minimum for n = 2, 3, 4, and 5 corresponds to the g+g-t, g+g+g-t, g-g+g+g-t, and g+g g+g+g-t conformations, respectively. Furthermore, the stability of the extended and semi-extended conformations increases systematically with the size of the central aliphatic segment. The relationship between the size of the aliphatic segment and the stability of folded conformations is analyzed and discussed. PMID- 15822954 TI - Synthesis of diversely functionalized hexahydropyrrolo[2,3-b]indoles using domino reactions, olefination, isomerization and Claisen rearrangement followed by reductive cyclization. AB - Hexahydropyrrolo[2,3-b]indoles 6 were synthesized in five steps from indolin-3 one 8 by a general and efficient method, in which elements of molecular diversity were readily added onto the 3a-position of the pyrrolo[2,3-b]indole ring system. Horner-Wadsworth-Emmons reaction of 2-allyloxyindolin-3-ones 7, derived from indolin-3-one 8 and a variety of allylic alcohols, smoothly proceeded with successive Claisen rearrangement to give the corresponding 3-allyl-3 cyanomethylindolin-2-ones 15. Indolin-2-ones 15 were converted into pyrrolo[2,3 b]indoles 6 using partial hydrolysis followed by reductive cyclization with LiAlH(4). Synthesis of N-methylated pyrrolo[2,3-b]indole derivatives 23 and 26 is also described. PMID- 15822955 TI - Ru(II)-SDP-complex-catalyzed asymmetric hydrogenation of ketones. Effect of the alkali metal cation in the reaction. AB - The Ru(II) complexes of SDP and DPEN combined with t-BuOK in 2-propanol formed a very effective catalyst for the hydrogenation of simple aromatic ketones with high activity and enantioselectivity. The racemic alpha-arylcycloalkanones can also be hydrogenated by this system, providing alpha-arylcycloalkanols in excellent cis/trans stereoselectivity (>99:1) and enantioselectivity (up to 99.9%) for the cis isomer. In the study of the effect of the alkali metal cation in the hydrogenation of acetophenone using RuCl(2)(Tol-SDP)(DPEN) and RuCl(2)(Xyl SDP)(DPEN) catalysts, we found that t-BuONa provided a faster reaction than t BuOK, which indicated that the sterically hindered diphosphine ligands preferred the base with the smaller metal cation. PMID- 15822956 TI - Convergent synthesis of nonsymmetric pi-stacked protophanes assembled with urea linkers. AB - A convergent approach has been developed for the preparation of nonsymmetric tertiary arylureas possessing two or three urea linkages. This approach has been used for the preparation of ureas possessing 1-naphthylenyl and 4-nitrophenyl end groups separated by either one or two phenylene diamine bridging units. These ureas were constructed as prototypes for donor-bridge-acceptor systems based on tertiary arylurea architecture. AM1 calculations indicate a preference of these arylureas for folded, protophane structures in which the aryl groups are loosely pi-stacked. Analysis of the (1)H NMR chemical shifts supports the assignment of folded structures in solution. The absorption and luminescence spectra of tertiary ureas possessing 1-naphthylenyl and/or 4-nitrophenyl are reported. The absence of fluorescence and appearance of structured phosphorescence at 77 K are attributed to nitrophenyl-localized lowest energy singlet and triplet states. Localization of excitation on the acceptor chromophore precludes investigation of charge transfer in these systems. PMID- 15822957 TI - Conformational rigidification via derivatization facilitates the determination of absolute configuration using chiroptical spectroscopy: a case study of the chiral alcohol endo-borneol. AB - We demonstrate that derivatization of the OH group of endo-borneol, 1, leads to conformational rigidification. Conformational analysis (CA) of 1 and its methyl, acetate, tert-butyl, and trimethylsilyl derivatives, 2-5, is carried out using ab initio density functional theory (DFT). The number of thermally accessible stable conformations is reduced from 3 in 1, to 2 in 2, and to 1 in 3-5. Comparison of IR and vibrational circular dichroism (VCD) spectra of 1 and 3-5, calculated using DFT, to experimental spectra unambiguously confirms the DFT CA. The determination of absolute configurations (ACs) of chiral molecules via analysis of chiroptical spectra using DFT methods increases in complexity and decreases in reliability as the number of populated conformations increases. Our results for endo-borneol support the conclusion that, in the case of chiral alcohols, derivatization can lead to substantial rigidification and, as a result, significantly facilitate the determination of ACs. PMID- 15822958 TI - The three corrugated surfaces of 1,4-divinyltetramethylene diradical intermediates and their connections to 1,2-divinylcyclobutane, 4 vinylcyclohexene, 1,5-cyclooctadiene, and two butadienes. AB - The three potential energy surfaces of the trans-trans, cis-trans, and cis-cis divinyltetramethylene diradicals have been located with DFT calculations at the BPW91/6-311+G levels. The three surfaces account well for the experimental results reported for the thermolysis of optically active trans-1,2 divinylcyclobutane and optically active and deuterated 4-vinylcyclohexene. The surfaces account also for the outcome of the dimerization of butadiene and the thermolysis of cis,cis-1,5-cyclooctadiene. The three diradical intermediates are connected to the cyclization and dissociation products through conformations that are explored fully here. PMID- 15822959 TI - Theoretical investigation of optical and electronic property modulations of pi conjugated polymers based on the electron-rich 3,6-dimethoxy-fluorene unit. AB - Poly(fluorene)-type materials are widely used in polymer-based emitting devices. One of the drawbacks of light-emitting diodes based on polyfluorene derivatives is the injection of holes from the anode due to the high ionization potential (IP) of most derivatives. Substitution by electron-donating alkoxy substituents or by adding charge carriers on the conjugated polymer's backbone produces a remarkable influence on its electrical and optical properties. In this contribution, we apply quantum-chemical techniques to investigate a family of pi conjugated polymers with substituted dimethoxy groups at the 3,6 positions of the fluorene ring, namely, poly(2,7-(3,6-dimethoxy-fluorene)(PDMOF), poly(2,7-(3,6 dimethoxy-fluorene)-co-alt-fluorene (PDMOFF), and poly(2,7-(3,6-dimeth-oxy fluorene)-co-alt-2,5-thiophene (PDMOFT). The electronic properties of the neutral molecules, HOMO-LUMO gaps (Delta(H)(-)(L)), in addition to the positive and negative ions, are studied using the B3LYP functional. The lowest excitation energies (E(g)) and the maximal absorption wavelength lambda(abs) of PDMOF, PDMOFF, and PDMOFT are studied by employing time-dependent density functional theory (TD-DFT) and the ZINDO semiempirical method. The IP, EA, and E(g) values of each polymer were obtained by extrapolating those of the oligomers to the inverse chain length equal to zero ((1)/(n)() = 0). The influence of the presence of methoxy groups on the fluorene moiety on the ionization potential is especially emphasized. The outcomes show that the HOMO energies of these systems under study increase by about 0.4 eV and the IP values decrease by about 0.3 eV compared to those of the corresponding polyfluorene. Both effects result in a reduction of the energy barrier for the injection of holes in related polymeric light-emitting devices and should contribute to the enhancement of their performances. Because of the cooperation with thiophene in PDMOFT, which results in a good planar conformation, both the hole-creating and electron-accepting abilities are improved. PMID- 15822960 TI - Practical asymmetric synthesis of a potent PDE4 inhibitor via stereoselective enolate alkylation of a chiral aryl-heteroaryl secondary tosylate. AB - A practical, chromatography-free catalytic asymmetric synthesis of a potent and selective PDE4 inhibitor (L-869,298, 1) is described. Catalytic asymmetric hydrogenation of thiazole ketone 5a afforded the corresponding alcohol 3b in excellent enantioselectivity (up to 99.4% ee). Activation of alcohol 3b via formation of the corresponding p-toluenesulfonate followed by an unprecedented displacement with the lithium enolate of ethyl 3-pyridylacetate N-oxide 4a generated the required chiral trisubstituted methane. The displacement reaction proceeded with inversion of configuration and without loss of optical purity. Conversion of esters 2b to 1 was accomplished via a one-pot deprotection, saponification, and decarboxylation sequence in excellent overall yield. PMID- 15822961 TI - Amphimelibiosides A-F, six new ceramide dihexosides isolated from a Japanese marine sponge Amphimedon sp. AB - Six new ceramide dihexosides, amphimelibiosides A-F (1-6), were isolated from a Japanese marine sponge Amphimedon sp. The structure of amphimelibioside C (3), which is a major component of amphimelibiosides, was determined by 2D NMR techniques, chemical degradation, and a semisynthetic method to be 1-O-[beta-D glucopyranosyl-(1-->6)-alpha-D-galactopyranosyl]-(2S,3S,4R,6E)-2-[(2'R)-2 hydroxydocosanoyl]-2-amino-6-octadecene-1,3,4-triol. The structures of the other constituents were elucidated by a combination of mass spectra, (1)H NMR, and GC MS analysis. PMID- 15822962 TI - Selective functionalization of 2-fluoropyridine, 2,3-difluoropyridine, and 2,5 difluoropyridine at each vacant position. AB - The concept of "regioexhaustive substitution" has been successfully applied to 2 fluoro-, 2,3-difluoro-, and 2,5-difluoropyridine. All vacant positions were amenable to regioselective metalation and subsequent carboxylation by employing either chlorine as a neighboring site activating protective group or trimethylsilyl as a neighboring site screening protective group. In this way, approximately half a dozen fluorinated pyridinecarboxylic acids were derived from each starting material. PMID- 15822963 TI - Thermal reactions of tributyltin hydride with alpha-azido esters: unexpected intervention of tin triazene adducts under both nonradical and radical conditions. AB - Thermal reaction of various alpha-azido esters with Bu(3)SnH in refluxing benzene results in smooth production of 3-(tributylstannyl)-1-triazene adducts affording cyclized 1,2,3-triazol-4-ones in preference to reduced amines and thence provides a new useful method for the preparation of these triazole derivatives. In the presence of AIBN the occurrence of triazene products still remains important or even exclusive and, consequently, generation of the expected stannylaminyl radicals is seriously limited. With 2-azidomalonates and alpha-azido-beta-keto esters stannyltriazenes can similarly occur in the absence of the radical initiator, but in the latter cases the ensuing triazenes undergo preferential cyclization onto the ketone moiety to give reactive hydroxytriazolines. Contrary to alpha-azido esters, in the presence of AIBN alpha-azido-beta-keto esters as well as azidomalonates give rise only to the usual stannylaminyl radicals. A possible explanation for the different behavior of the mono- and dicarbonyl azides in the presence of AIBN is put forward. PMID- 15822964 TI - Flexible synthesis of enantiomerically pure 2,8-dialkyl-1,7 dioxaspiro[5.5]undecanes and 2,7-dialkyl-1,6-dioxaspiro[4.5]decanes from propargylic and homopropargylic alcohols. AB - A new approach to enantiomerically pure 2,8-dialkyl-1,7-dioxaspiro[5.5]undecanes and 2,7-dialkyl-1,6-dioxaspiro[4.5]decanes is described and utilizes enantiomerically pure homopropargylic alcohols obtained from lithium acetylide opening of enantiomerically pure epoxides, which are, in turn, acquired by hydrolytic kinetic resolution of the corresponding racemic epoxides. Alkyne carboxylation and conversion to the Weinreb amide may be followed by triple-bond manipulation prior to reaction with a second alkynyllithium derived from a homo- or propargylic alcohol. In this way, the two ring components of the spiroacetal are individually constructed, with deprotection and cyclization affording the spiroacetal. The procedure is illustrated by acquisition of (2S,5R,7S) and (2R,5R,7S)-2-n-butyl-7-methyl-1,6-dioxaspiro[4.5]-decanes (1), (2S,6R,8S)-2 methyl-8-n-pentyl-1,7-dioxaspiro[5.5]undecane (2), and (2S,6R,8S)-2-methyl-8-n propyl-1,7-dioxaspiro[5.5]undecane (3). The widely distributed insect component, (2S,6R,8S)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane (4), was acquired by linking two identical alkyne precursors via ethyl formate. In addition, [(2)H(4)] regioisomers, 10,10,11,11-[(2)H(4)] and 4,4,5,5-[(2)H(4)] of 3 and 4,4,5,5 [(2)H(4)]-4, were acquired by triple-bond deuteration, using deuterium gas and Wilkinson's catalyst. This alkyne-based approach is, in principle, applicable to more complex spiroacetal systems not only by use of more elaborate alkynes but also by triple-bond functionalization during the general sequence. PMID- 15822965 TI - Conversion of five-, six-, and seven-membered lactams to racemic or scalemic 2 substituted heterocycles by amidoalkylation. AB - An efficient method for the synthesis of 2-alkyl- and 2-aryl pyrrolidines, piperidines, and azepanes from lactams, in either racemic or enantiopure form, is presented. The lactam nitrogens are acylated with either Boc anhydride or trans cumylcyclohexyl (TCC) chloroformate. Selective reduction of the lactam carbonyl to the carbinolamide is followed by treatment with benzotriazole. Substitution of the benzotriazole is accomplished by treatment with organometallics, yielding the 2-substituted heterocycles. With TCC, up to 90% diastereoselectivity is achieved. After diastereomer purification, reductive removal of the auxiliary affords enantiopure 2-substituted heterocycles. A mechanistic hypothesis is presented that details the conformational equilibria of the key step. PMID- 15822966 TI - Synthesis of 2,2'-biimidazolium-based ionic liquids: use as a new reaction medium and ligand for palladium-catalyzed suzuki cross-coupling reactions. AB - Neat reactions of 2,2'-biimidazole with an excess of alkyl or polyfluoroalkyl iodides at 140 degrees C, followed by anion exchange with LiN(SO(2)CF(3))(2) or KPF(6), gave the diquaternary salts 3a-k in >80% yields. However, by controlling the reaction stoichiometry, 2,2'-biimidazole can also be monoquaternized with the same electrophiles at 100 degrees C under similar conditions. Subsequent metathesis reactions with LiN(SO(2)CF(3))(2) or KPF(6) resulted in the ionic liquids 4a-m in high yields. Thermal properties were determined with a differential scanning calorimeter (DSC) and a thermogravimetric analyzer (TGA). Most of the monoquaternary salts are room-temperature ionic liquids. 1,3,1' Tributyl-2,2'-biimidazolium hexafluorophosphate was demonstrated to be an excellent solvent and ligand for palladium-catalyzed Suzuki cross-coupling reactions. The catalytic ionic liquid system may be recycled at least 14 times without a significant decrease in catalytic performance. PMID- 15822968 TI - Kinetics and mechanism of the addition of benzylamines to benzylidene-3,5 heptadione in acetonitrile. AB - The addition reaction of benzylamine (XC6H4CH2NH2) to benzylidene-3,5-heptadione (BHD; YC6H(4-)CH=C(COEt)(2)) in acetonitrile is investigated. The rate is slower than the corresponding rate for benzylidenediethylmalonate (YC6H4CH=C(OOEt)(2)) as the result of a greater steric hindrance in the planar dicarbonyl transition state. The kinetic isotope effects (k(H)/k(D)) involving deuterated amine nucleophiles (XC6H4CH2ND2) are greater than 1 (1.37-2.04), indicating N-H bond stretching with concurrent N-C(alpha) and H-C(beta) bond formation in the TS. The trend of change in k(H)/k(D) with variation of substituent X in the nucleophile conforms to the Bell-Evans-Polanyi principle. It has been stressed that the dicarbonyl group activated olefins exhibit insignificant charge imbalance in the TS for the benzylamine additions in acetonitrile as a result of the two strong n(c) --> pi*(C=O) vicinal charge-transfer interactions. PMID- 15822967 TI - Total synthesis of bassiatin and its stereoisomers: novel divergent behavior of substrates in Mitsunobu cyclizations. AB - Total syntheses of the morpholine-2,5-dione, Bassiatin, and its stereoisomers have been completed. A key step in the syntheses was the Mitsunobu cyclization of hydroxyacid acyclic precursors. The hydroxyacid precursors are hindered alcohols and two substrates underwent Mitsunobu cyclization with retention of configuration. The other two substrates underwent Mitsunobu cyclization with either retention or inversion of configuration depending on reaction conditions. This divergence in outcome of the Mitsunobu reaction for the same substrate depending on effective concentration is novel. PMID- 15822969 TI - Easy-to-execute carbonylations: microwave synthesis of acyl sulfonamides using Mo(CO)(6) as a solid carbon monoxide source. AB - The development of a robust palladium-catalyzed amidocarbonylation protocol for the preparation of aromatic acyl sulfonamides utilizing high-density microwave heating is described. This synthetic approach employs Mo(CO)(6) as a convenient CO-releasing reagent and allows for the direct preparation of acyl sulfonamides from both aryl iodides and aryl bromides. The reactions can be performed under air, employing only 15 min of microwave irradiation, to produce acyl sulfonamide derivatives in good to excellent yields. To illustrate the usefulness of this method, we reported the synthesis of a novel hepatitis C virus NS3 protease inhibitor. PMID- 15822970 TI - Palladium-catalyzed synthesis of tetrahydrofurans from gamma-hydroxy terminal alkenes: scope, limitations, and stereoselectivity. AB - A new, stereoselective synthesis of substituted tetrahydrofurans via Pd-catalyzed reactions of aryl and vinyl bromides with gamma-hydroxy terminal alkenes is described. This transformation affords trans-2,5- and trans-2,3-disubstituted tetrahydrofurans with up to >20:1 dr. This methodology also provides access to bicyclic and spirocyclic tetrahydrofuran derivatives in good yield with 10-20:1 dr. The scope and limitations of these transformations are discussed in detail, as are the effect of substrate sterics and electronics on yield and stereoselectivity. A proposed mechanism of these transformations is presented along with a model that rationalizes the stereochemical outcome of the reactions. PMID- 15822971 TI - Conventional and microwave-assisted conversion of substituted 3-amino-oxazolidin 2,4-diones into N',N'-disubstituted alpha-hydroxyhydrazides. AB - Substituted 3-amino-oxazolidin-2,4-diones have been prepared by reacting cyanohydrins or alpha-hydroxyesters subsequently with 1,1'-carbonyldiimidazole and 1,1-disubstituted hydrazines followed by acidic hydrolysis in case of the intermediate 3-amino-4-imino-oxazolidin-2-ones. Conventional and microwave assisted syntheses of N',N'-disubstituted alpha-hydroxyhydrazides have been accomplished by reacting substituted 3-amino-oxazolidin-2,4-diones with catalytic amounts of sodium methoxide in methanol. PMID- 15822972 TI - Ortho-selective direct cross-coupling reaction of 2-aryloxazolines and 2 arylimidazolines with aryl and alkenyl halides catalyzed by ruthenium complexes. AB - The ortho position of the aromatic ring in 2-aryloxazolines and 2 arylimidazolines is selectively arylated and alkenylated with organic halides in the presence of a ruthenium(II)-phosphine complex. In the case of unsubstituted and para-substituted phenyloxazolines, 1:2 coupled products were obtained preferentially, while 1:1 coupled products were obtained in the case of meta substituted phenyloxazolines and N-acylarylimidazolines. The reaction is proposed to proceed via the generation of an organoruthenium intermediate, formed by oxidative addition of the organic halide, and ortho-ruthenation directed by the coordination of the 2-oxazolinyl or 2-imidazolinyl group to the ruthenium center. PMID- 15822973 TI - Stereoselective synthesis of four stereoisomers of beta-methoxytyrosine, a component of callipeltin A. AB - Callipeltin A is a novel cyclic depsipeptide that selectively inhibits the cardiac sodium/calcium exchanger and is therefore of interest as a regulator of myocardial contractility. The stereochemistry of beta-methoxytyrosine, one of the amino acids contained within the marine natural product, could not be determined. In an effort to elucidate the stereochemistry of this moiety, a stereoselective synthesis of four stereoisomers of beta-methoxytyrosine from (S)- and (R)-serine was accomplished. PMID- 15822974 TI - Photogenerated N-methyl-N-1-naphthylnitrenium ion: laser flash photolysis, trapping rates, and product study. AB - N-Methyl-N-1-naphthylnitrenium ion (2) was generated through photolysis of 1-(N methyl-N-(1-naphthyl)amino)-2,4,6-trimethylpyridinium tetrafluoroborate (1). Laser flash photolysis (LFP) with time-resolved UV-vis (TRUV) detection as well as photoproduct analysis verified that the expected nitrenium ion was formed cleanly and rapidly following photolysis. Consistent with an earlier study, which used competitive trapping methods (Novak, M. et al. J. Org. Chem. 1999, 64, 6023 6031), it is found that 2 reacts rapidly with a variety of nucleophiles. The high reactivity of 2 relative to other arylnitrenium ions is discussed in terms of steric and electronic effects. PMID- 15822975 TI - The wewakpeptins, cyclic depsipeptides from a Papua New Guinea collection of the marine cyanobacterium Lyngbya semiplena. AB - Four new depsipeptides have been isolated from the marine cyanobacterium Lyngbya semiplena collected from Papua New Guinea. The amino and hydroxy acid partial structures of wewakpeptins A-D (1-4) were elucidated through extensive spectroscopic techniques, including HR-FABMS, 1D (1)H and (13)C NMR, as well as 2D COSY, HSQC, HSQC-TOCSY, and HMBC spectra. The sequence of the residues was determined through a combination of multifaceted approaches including ESI-MS/MS, HMBC, ROESY, and a modified 1D HMBC experiment. The absolute stereochemistry of each residue was determined by chiral HPLC and chiral GC-MS methods. The wewakpeptins represent an unusual arrangement of amino and hydroxy acid subunits relative to known cyanobacterial peptides and possess a bis-ester, a 2,2-dimethyl 3-hydroxy-7-octynoic acid (Dhoya) or 2,2-dimethyl-3-hydroxyoctanoic acid (Dhoaa) residue, and a diprolyl group reminiscent of dolastatin 15. Wewakpeptin A and B were the most cytotoxic among these four depsipeptides with an LC(50) of approximately 0.4 muM to both the NCI-H460 human lung tumor and the neuro-2a mouse neuroblastoma cell lines. PMID- 15822976 TI - Oxidative formation of thiolesters in a model system of the pyruvate dehydrogenase complex. AB - In the presence of a catalytic amount of 3-butyl-4-methylthiazolium bromide, the reaction of benzaldehydes with azobenzene in dichloromethane containing octanethiol and Et(3)N gave the corresponding S-octyl thiobenzoates in good yields. The thiolesters were produced by trapping of the 2-benzoylthiazolium salts with the thiol, which were generated through the azobenzene oxidation of the active aldehydes. This is the first example for the thiolester formation mimicking the function of the pyruvate dehydrogenase complex. An electron withdrawing substituent at the 4-position of benzaldehyde enhanced the reaction rate. The effect of benzaldehyde substituents on the reaction rate was examined quantitatively on the basis of kinetic measurements, leading to a nonlinear correlation of log(k(obs)) with Hammett's substituent constants (sigma). The origin of the nonlinear Hammett plot was interpreted in terms of a shift in the rate-determining step of the multistep reaction with change of the electronic nature of substituent. Further support for this assumption was given by the observation that the reaction constant (rho) of the Hammett plot for the azobenzene substituent effect on the oxidation rate of 4-bromobenzaldehyde was much smaller than that of 4-cyanobenzaldehyde. PMID- 15822977 TI - Metalloporphyrin-capped calix[4]pyrroles: heteroditopic receptor models for anion recognition and ligand fixation. AB - Diametrically strapped calix[4]pyrrole-metalloporphyrin conjugates, potential hosts for anionic guests, have been synthesized and characterized. The syntheses rely on the acid-catalyzed condensation of two dipyrromethane bearing Ni(II) porphyrins with acetone. An (1)H NMR spectroscopic titration experiment indicated that the resulting receptors selectively trapped fluoride anions in organic media but not other, larger halide anions. The experimental results from titration and Job plots indicated that the bound fluoride anion must reside inside the cavity. The current systems provide a well-defined illustration of how size-selective anion receptors may be synthesized by incorporating recognition functions, such as Lewis acidity, hydrogen bonding, and encapsulating moieties into established recognition motifs, calix[4]pyrrole in the present instance. PMID- 15822978 TI - Asymmetric synthesis of the azabicyclic core of the Stemona alkaloids. AB - A general strategy for the construction of the 1-azabicyclo[5.3.0]decane core of Stemona alkaloids is developed. Our diversity-oriented approach exploits 1,3 dipolar cycloaddition of five-membered cyclic nitrones to C(6) olefins, followed by N-O reductive cleavage and azepine closure. The use of various enantiopure pyrroline N-oxides allows for a practical, stereoselective preparation of several putative precursors of different Stemona alkaloids. PMID- 15822979 TI - Solution syntheses of protected type II Lewis blood group oligosaccharides: study for automated synthesis. AB - Glycosyl phosphate and trichloroacetimidate monosaccharide building blocks were used in a stepwise solution-phase synthesis of three Lewis blood group oligosaccharides. The syntheses were conducted to establish general routes for the automated assembly of the oligosaccharide portion of biologically important glycolipids. The H-type II pentasaccharide, Le(x) pentasaccharide, and Le(y) hexasaccharide were prepared in high yield. These syntheses served to evaluate the utility and limitations of the 2-(azidomethyl)benzoate ester (AZMB) for the construction of complex carbohydrates. Development of a glucosamine building block containing a N-trichloroacetamide group to mask the C2 amine improved coupling yields and was key for completion of the Le(x) and Le(y) structures. PMID- 15822980 TI - Direct, two-step synthetic pathway to novel dibenzo[a,c]phenanthridines. AB - Novel dibenzo[a,c]phenanthridines are prepared regioselectively by the application of a straightforward synthetic pathway, starting from new 3,4-diaryl- and 3,4-dihydro-3,4-diarylisoquinolines prepared via Ritter-type heterocyclization and the more classical two-step reductive amination/Bischler Napieralski cyclization of triarylethanones, respectively. A comparative study of nonphenolic oxidative coupling methodologies provides a highly efficient procedure, based on the hypervalent iodine reagent phenyliodine(III) bis(trifluoroacetate) (PIFA), to accomplish the final coupling step. PMID- 15822981 TI - "Tethered" Ru(II) catalysts for asymmetric transfer hydrogenation of ketones. AB - Stereochemically well-defined ruthenium(II) catalysts have been applied to the asymmetric transfer hydrogenation of a series of ketones. In one case, statistical experimental design was employed to optimize the enantiomeric excess of the product. In the case of the TsDPEN-based systems, the replacement of trans 1,2-diphenyl substitution with cis-, or deletion of one of the phenyl groups, results in significant deterioration of the enantiomeric excess. A new method is described for the synthesis of tethered amino alcohol-containing catalysts. PMID- 15822982 TI - Metal-mediated entry to functionalized 3-substituted 3-hydroxyindolin-2-ones via regiocontrolled carbonylallylation, bromoallylation, 1,3-butadien-2-ylation, propargylation, or allenylation reactions of isatins in aqueous media. AB - Reactions of 2,3-indolinediones (isatins) with stabilized organometallic reagents were investigated in aqueous media. Isatins and a variety of stabilized organic halides undergo coupling under Barbier-type conditions in the presence of different metals (indium, tin, zinc) and additives (ammonium chloride, hydrobromic acid, bismuth(III) chloride, hafnium(IV) chloride). The regiochemistry of the processes (carbonylallylation, bromoallylation, 1,3 butadien-2-ylation, propargylation, or allenylation reactions) were generally excellent. On this basis, simple and fast protocols for the synthesis of the potentially bioactive 3-substituted 3-hydroxyoxindole moiety were developed. PMID- 15822983 TI - Fluorinated olefinic peptide nucleic acid: synthesis and pairing properties with complementary DNA. AB - The fluorinated olefinic peptide nucleic acid (F-OPA) system was designed as a peptide nucleic acid (PNA) analogue in which the base carrying amide moiety was replaced by an isostructural and isoelectrostatic fluorinated C-C double bond, locking the nucleobases in one of the two possible rotameric forms. By comparison of the base-pairing properties of this analogue with its nonfluorinated analogue OPA and PNA, we aimed at a closer understanding of the role of this amide function in complementary DNA recognition. Here we present the synthesis of the F OPA monomer building blocks containing the nucleobases A, T, and G according to the MMTr/Acyl protecting group scheme. Key steps are a selective desymmetrization of the double bond in the monomer precursor via lactonization as well as a highly regioselective Mitsunobu reaction for the introduction of the bases. PNA decamers containing single F-OPA mutations and fully modified F-OPA decamers and pentadecamers containing the bases A and T were synthesized by solid-phase peptide chemistry, and their hybridization properties with complementary parallel and antiparallel DNA were assessed by UV melting curves and CD spectroscopic methods. The stability of the duplexes formed by the decamers containing single (Z)-F-OPA modifications with parallel and antiparallel DNA was found to be strongly dependent on their position in the sequence with T(m) values ranging from +2.4 to -8.1 degrees C/modification as compared to PNA. Fully modified F-OPA decamers and pentadecamers were found to form parallel duplexes with complementary DNA with reduced stability compared to PNA or OPA. An asymmetric F OPA pentadecamer was found to form a stable self-complex (T(m) approximately 65 degrees C) of unknown structure. The generally reduced affinity to DNA may therefore be due to an increased propensity for self-aggregation. PMID- 15822984 TI - A novel efficient deoxygenation process for N-heteroarene N-oxides. AB - A novel deoxygenation process for N-heteroarene N-oxides is described. The deoxygenation process has been carried out by utilizing some short C-chain alcohols, benzyl alcohol, or 1-phenylethanol as the solvent in the presence of a base, such as sodium alkoxide or sodium hydroxide. A series of N-heteroarene N oxides was submitted to the developed conditions to provide the corresponding N heteroarenes with high yield and excellent selectivity. When the deoxygenation is carried out with benzyl alcohol or 1-phenylethanol as the reaction medium, the process can be performed under very mild conditions, at only 30 degrees C. The deoxygenation process is in contrast to several other methods performed without the presence of any transition metal as a catalyst or stoichiometric reagent. DFT calculations suggest that the alkoxide performs a nucleophilic attack on the N heteroarene in the ortho or para position. This bond is cleaved homolytically with the overall result being that a single electron-transfer step has occurred. The products of this process are an N-heteroarene N-oxide radical anion and an alkoxyl or benzyloxy radical, depending on the solvent that has been used. Successive steps of the mechanism result in an oxygen transfer from the N-oxide to give the deoxygenated N-heteroarene and 1 equiv of the aldehyde, which is the oxidation product of the solvent alcohol. PMID- 15822985 TI - N-nitrosoamide-mediated N --> O nitroso transfer to alcohols with subsequent denitroxylation. AB - Decomposition of certain N-benzyl-N-nitrosoamides is often accompanied by small amounts of benzaldehyde whose formation was postulated to arise from in situ formation and oxidation of benzyl alcohol. Incubation of excess benzyl alcohol with thermostable N-benzyl-N-nitrosoamides at ambient temperatures in inert solvents generates benzyl nitrite, N-benzyl amides, and benzaldehyde as the major products. Benzyl nitrite formation appears to be linked to N --> O nitroso transfer between the N-benzyl-N-nitrosoamides and benzyl alcohol, which is subject to the previously observed electronic and steric features of the acyl substituent although the former appears to play a much larger role than the latter. Benzaldehyde formation evidently arises from dehydronitrosation (denitroxylation) of the nitrite via O-N bond homolysis and H-abstraction from the resultant benzyloxy radical. Although trans-nitrosation occurs with methanol, 1 degree, 2 degree, and 3 degree alcohols, the reaction is evidently subject to steric effects at both the alpha and beta carbons of the alcohol. Additionally, carbonyl formation only occurs with 2 degrees alcohols and those that can derive resonance-stabilized carbonyls. PMID- 15822986 TI - Bis(m-phenylene)-32-crown-10-based cryptands, powerful hosts for paraquat derivatives. AB - Four new bis(m-phenylene)-32-crown-10-based cryptands with different third bridges were prepared. Their complexes with paraquat derivatives were studied by proton NMR spectroscopy, mass spectrometry, and X-ray analysis. It was found that these cryptands bind paraquat derivatives very strongly. Specifically, a diester cryptand with a pyridyl nitrogen atom located at a site occupied by either water or a PF(6) anion in analogous complexes exhibited the highest association constant K(a) = 5.0 x 10(6) M(-1) in acetone with paraquat, 9000 times greater than the crown ether system. X-ray structures of this and analogous complexes demonstrate that improved complexation with this host is a consequence of preorganization, adequate ring size for occupation by the guest, and the proper location of the pyridyl N-atom for binding to the beta-pyridinium hydrogens of the paraquat guests. This readily accessible cryptand is one of the most powerful hosts reported for paraquats. PMID- 15822987 TI - A computational study of [2.2]cyclophanes. AB - A computational study of isomeric [2.2]cyclophanes, namely [2.2]paracyclophane 1, [2.2]metacyclophane 2, and [2.2]metaparacyclophane 3, has been carried out. For 1, geometry optimizations performed by various methods at different basis sets showed that MP2/6-31+G(d,p) and B3PW91/6-31+G(d,p) provide the best results in comparison to the X-ray data. Compound 1 has D(2) symmetry with distorted bridges. A conformational search was performed for [2.2]cyclophanes 2 and 3. Each cyclophane exists in two conformations which have different energies in the case of 3 but are degenerate in the case of 2. Relative energies and strain energies at the bridges follow the same order, indicating that the relief of bridge tension and repulsion between pi clouds are determining factors for the stability of [2.2]cyclophanes. Through a decomposition of strain energy, it can be concluded that both the rings or the bridges can absorb strain, but it depends on the conformer of butane that is considered in the calculation of SE(br). Changes in aromaticity of these compounds were evaluated by NICS and HOMA and were compared with benzene and xylenes dimers as models. Despite distortions from planarity and shortening and lengthening of the C-C bonds relative to the mean, the phenyl rings are aromatic. NICS suggests a concentration of electronic density between the rings as a result of bridging process. Computed MK, NPA, and GAPT charges were compared for the isomeric cyclophanes. The GIAO chemical shifts were calculated and indicate that 1 has a larger diamagnetic anisotropy than the other isomers. PMID- 15822988 TI - Ionic-liquid-supported peptide synthesis demonstrated by the synthesis of Leu(5) enkephalin. AB - A new strategy for the synthesis of oligopeptides was developed using an ionic liquid as a soluble support. The efficiency of this ionic liquid-phase approach was demonstrated by the synthesis of a bioactive pentapeptide, Leu(5)-enkephalin, in good yield and reasonable purity. The structures and purities of the reaction intermediates in each step were verified easily by routine spectroscopic analysis, and no chromatographic procedures were needed during the synthesis. PMID- 15822989 TI - Theoretical study of the highly diastereoselective 1,3-dipolar cycloaddition of 1,4-dihydropyridine-containing azomethine ylides to [60]fullerene (Prato's reaction). AB - The 1,3-dipolar cycloaddition of azomethine ylides bearing the biologically active 1,4-dihydropiridine ring to C(60) was investigated by means of quantum mechanical calculations at the semiempirical AM1 and DFT (B3LYP/6-31G) methods. The presence of two chiral centers and one chiral axis in the resulting fulleropyrrolidines leads to four possible [6,6] cycloaddition products. Formation of atropoisomers has also been considered. The transition-state structures were computed for the four different cycloaddition pathways to find out the lowest activation energy stereoisomer. In all cases, a frequency analysis and an IRC calculation were carried out to fully characterize the located transition-state structures. AM1 results and single-point energy calculations at the B3LYP/6-31G//AM1 level for the four transition-state structures yield activation energies values below 5 kcal/mol. PMID- 15822990 TI - Isolation and X-ray structures of labile benzoic- and acetic-acidium carbocations. AB - New carbocationic salts (via O-protonation of substituted benzoic acids) are prepared for the first time by controlled hydration of the corresponding benzoylium salts and isolated in pure crystalline form. Precise X-ray structural analyses reveal the rather unexpected (electronic) structure of the carboxylic acidium functionality. PMID- 15822991 TI - Synthesis and structural study of thiacyclophanes utilizing dibromides and methane dithiolate. AB - The synthesis of a series of thiacyclophanes and optically active binaphthol based chiral thiacyclophanes is reported with XRD structure. Two diastereomeric tetrathiacyclophanes are designed and synthesized. The two diastereomers are evidenced by crystal structure; the single-crystal X-ray studies reveal that one of the isomers possesses an inherent property of self-assembling into a vertical stack of tunnel-like structures. PMID- 15822992 TI - Synthesis of heteroaryl 1,2-diketones. AB - Reactions of lithiated heteroaryl- (or aryl-) methylbenzotriazoles 4a-e with heteroaryl (or aryl) esters 5a-f give alpha-benzotriazolyl ketones 6a-n in average yields of 90-95%. Oxidation of ketones 6a-l in aqueous DMSO in the presence of oxygen and sodium bicarbonate gave heteroaryl- (aryl-) 1,2 ethanediones 7a-l in average yields of 60-80%. PMID- 15822993 TI - Total synthesis of alternariol. AB - Total synthesis of alternariol, a toxic secondary metabolite of various Alternaria fungi, was achieved in seven steps starting with orcinol and 3,5 dimethoxybromobenzene. The longest linear sequence consists of six steps. Key reaction is a palladium-catalyzed Suzuki-type coupling of an orcinol-derived boronic acid with a brominated resorcylic aldehyde. The final demethylation furnished alternariol in 73% yield containing a smaller fraction of alternariol 9 methyl ether (approximately 20%). PMID- 15822994 TI - A novel formal total synthesis of cephalotaxine. AB - A formal total synthesis of cephalotaxine (CET), the parent structure of antileukemia Cephalotaxus alkaloids, was achieved through a novel synthesis of the pentacyclic amino enone 4 by a rapid annulation of readily available beta (3,4-methylenedioxy)phenethylamine (2), delta-valerolactone, and bromoacetone. PMID- 15822996 TI - Efficient synthesis of tribenzohexadehydro[12]annulene and its derivatives in the ionic liquid. AB - A simple and efficient synthesis of tribenzohexadehydro[12]annulene and related derivatives in the ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate is described. A Sonogashira coupling reaction is the key step. In this system, the amount of CuI normally used can be reduced so that homocoupling is minimized. PMID- 15822995 TI - Application of vinyl tris(trimethylsilyl)germanes in Pd-catalyzed couplings. AB - The oxidative treatment of vinyltris(trimethylsilyl)germanes with hydrogen peroxide (NaOH/H(2)O/THF) or tert-butyl peroxide (KH/THF) generates reactive germanol or germanoxane species that undergo Pd-catalyzed cross-couplings with aryl and alkenyl halides and aryl triflates in the presence of Pd(PPh(3))(4). Vinylgermanes having either a conjugated or isolated double bond serve as versatile transmetalation reagents. The E-germanes undergo coupling with retention of stereochemistry under aqueous and anhydrous conditions, while coupling of Z-germanes occurs with less stereoselectivity to produce a mixture of E/Z products. PMID- 15822997 TI - Fluorinated heterocyclic compounds. An effective strategy for the synthesis of fluorinated Z-oximes of 3-perfluoroalkyl-6-phenyl-2h-1,2,4-triazin-5-ones via a ring-enlargement reaction of 3-benzoyl-5-perfluoroalkyl-1,2,4-oxadiazoles and hydrazine. AB - The reaction of 3-benzoyl-5-perfluoroalkyl-1,2,4-oxadiazoles with hydrazine has been investigated, evidencing the possibility of competitive reaction paths. Nucleophilic addition of the hydrazine to the electrophilic C(5) of the 1,2,4 oxadiazole ring, followed by ring opening and ring closure with enlargement, leads with high yield and in very mild experimental conditions to the formation of Z-oximes of 3-perfluoroalkyl-6-phenyl-2H-1,2,4-triazin-5-ones (11a-c) as major products of the reaction. In turn, the hydrazine can attack the electrophilic carbonyl carbon giving 4-perfluoroacylamino-5-phenyl-2H-1,2,3-triazoles (13a-c) through the well-known Boulton-Katritzky rearrangement of the intermediate hydrazones. PMID- 15822998 TI - Asymmetric cyclopropanation of 2,5-dimethyl-2,4-hexadiene by copper catalysts bearing new bisoxazoline ligands. AB - Some new bisoxazoline ligands with an aryl group at the 4-position and gem dimethyl groups at the 5-position on the oxazoline ring were prepared from arylglycines. Remarkable enhancement of the trans-selectivity (trans/cis = 87/13) and the enantioselectivity (96% ee for the trans product) was demonstrated for the asymmetric cyclopropanation of 2,5-dimethyl-2,4-hexadiene with tert-butyl diazoacetate catalyzed by the new copper-bisoxazoline complex. PMID- 15822999 TI - Aziridines versus vinyl carbamates from the direct amination of electron withdrawing group-substituted trifluoromethyl enoates. AB - Aza-MIRC (Michael-initiated ring closure) and C-H insertion products were obtained in the reactions of trifluoromethylated olefins with different nosyloxycarbamates by changing base and solvent. Aza-Michael addition products were not isolated. The presence and the position of the trifluoromethyl group allow control of the outcome of the reactions. PMID- 15823000 TI - An efficient synthesis of valienamine via ring-closing metathesis. AB - An efficient synthesis of valienamine is described. Valienamine was synthesized starting from commercially available 2,3,4,6-tetra-O-benzyl-D-glucose in nine steps, using ring-closing metathesis of (4S,5S,6S)-4,5,6-tribenzyloxy-7 (benzyloxymethyl)octa-1,7-dien-3-ol as a key step. PMID- 15823002 TI - Application of bis-acetylenic ketones in synthesis: one-pot preparation of the 1,2,4-triazepine and oxatriazaindenone cores. AB - Bis-acetylenic ketones were used to generate two heterocyclic templates. Two efficient one-pot procedures for the preparation of the triazepine and oxa triazaindenone cores are described. PMID- 15823001 TI - 31P NMR study of the desulfurization of oligonucleoside phosphorothioates effected by "aged" trichloroacetic acid solutions. AB - When employing phosphoramidites 1a-d in the solid-phase synthesis of oligonucleoside phosphorothioates, the thermolytic 2-[N-methyl-N-(2 pyridyl)]aminoethyl thiophosphate protecting group is lost to a large extent during the course of the synthesis. The resulting phosphorothioate diesters are then substantially desulfurized upon recurring exposure to a commercial solution of deblocking reagent during chain assembly. This problem is caused by the secondary decomposition product(s) of the reagent and is alleviated by using a fresh solution of the deblocking reagent prepared from solid trichloroacetic acid. PMID- 15823003 TI - Catalytic oxidation adjacent to carbonyl groups and at benzylic positions with a fluorous seleninic acid in the presence of iodoxybenzene. AB - In conjunction with iodoxybenzene as oxidant, perfluorooctylseleninic acid serves as a catalyst for the oxidation of aryl alkyl ketones to the corresponding ketoacids. Benzylic methylene groups are also oxidized by this reagent combination to the corresponding ketones. PMID- 15823004 TI - Sequential cyclization-elimination route to carbohydrate-based oxepines. AB - A five-step preparation of carbohydrate-based oxepines from hept-1-enitols is presented. The hept-1-enitols were subjected to silyl protection and hydroboration/oxidation to give the protected heptan-1-itols. Swern oxidation of the homologated alcohols followed by sequential acetal formation/cyclization provided methyl 2-deoxyseptanosides that underwent elimination reactions to give the carbohydrate-based oxepines. The new sequence is an alternative to the ring closing metathesis for the synthesis of carbohydrate-based oxepines from protected pyranose sugars. The product oxepines can serve as glycosyl donors in the synthesis of novel septanose carbohydrates. In addition, C-methylenealdehydo arabinofuranoside 16 was formed from 2-deoxyseptanoside 10 as the only product during protic acid mediated elimination reactions. This novel ring contraction complements other reported preparations of C-methylenaldehydo furanosides and underscores the acid-mediated reactivity introduced by competing eliminations between the C-1/C-2 and C-2/C-3 bonds. PMID- 15823005 TI - Highly functionalized cyclopentanes from meso bicyclic hydrazines. A rapid access to mannosidase inhibitors. AB - A simple diastereoselective access to amino- and hydrazinocyclopentitols is described. The key step involves a cationic rearrangement of a meso bicyclic hydrazine, followed by two successive stereoselective hydroxylations. Both racemic compounds are micromolar alpha-mannosidase (Jack bean) inhibitors. PMID- 15823006 TI - Conversion of furans into gamma-hydroxybutenolides: use of sodium chlorite. AB - 3,4-disubstituted furans are converted into gamma-hydroxy-butenolides by treatment with NaClO(2) in aqueous EtOH containing NaH(2)PO(4). PMID- 15823007 TI - Effect of water on the palladium-catalyzed amidation of aryl bromides. AB - The presence of water was found to have a significant impact on the conversion of some Hartwig-Buchwald reactions between amides with arylbromides. Seven amidation reactions were studied in a series of experiments containing 0-400 mol % water. When cesium carbonate is used as the base both in dioxane and toluene, water can have a beneficial effect. The experiments have also shown that for some reactions, anhydrous conditions are unnecessary and sometimes can hinder the reaction. PMID- 15823008 TI - Triethylborane-induced intermolecular radical addition to ketimines. AB - The intermolecular carbon radical addition to ketimines was investigated by using triethylborane as a radical initiator. The screening of reactive radical acceptors showed that pyruvic hydrazone 3 and isatin hydrazone 7 exhibit good reactivities toward nucleophilic alkyl radicals. The reaction of 3 and 7 proceeded effectively even under aqueous-medium reaction conditions. In the presence of BF(3).OEt(2), the radical addition to chiral ketimine 11 proceeded with good diastereoselectivities. PMID- 15823009 TI - Efficient syntheses of the keto-carotenoids canthaxanthin, astaxanthin, and astacene. AB - Three keto-carotenoids were prepared by the oxidation of the stable C(40) trisulfone 6, which has been used as the key compound in our beta-carotene synthesis. The first allylic oxidation to the unsaturated ketone and the second oxidation to the alpha-hydroxyketone produced the C(40) trisulfones 7 and 10, respectively. The Ramberg-Backlund reaction of the oxidized C(40) trisulfone was efficiently effected by the use of a mild base, NaOMe, in the presence of CCl(4) as a halogenating agent to give the C(40) disulfones 8 and 11. Base-promoted dehydrosulfonation reaction of the disulfone compounds produced the fully conjugated polyenes of canthaxanthin (1), astaxanthin (2), and astacene (3). PMID- 15823010 TI - An unusual intramolecular hetero-Diels-Alder cycloaddition. AB - A new reaction of erythronolides, an intramolecular hetero-Diels-Alder, has been discovered. Heated aqueous alcoholic solutions of ABT-773 (1) and its cis isomer (3) convert slowly to cycloadducts 2 and 4, respectively. Optimal reaction conditions, mechanistic studies supported by molecular modeling, and biological activity data are reported. Single-crystal X-ray structures for both adducts 2 and 4 have been obtained. PMID- 15823011 TI - Pd(0)-catalyzed highly selective synthesis of 1,1-diarylpropadienes and 1,3 diarylpropynes from 1-aryl-1-propynes and aryl halides. AB - Under different reaction conditions, 1,1-diarylpropadienes and 1,3-diarylpropynes can be prepared respectively by the sequential lithiation of 1-aryl-1-propynes, transmetalation, and the corresponding Pd(0)-catalyzed cross-coupling with aryl halides. A rationale for the formation of 1,3-diarylpropynes is proposed. PMID- 15823012 TI - cis-chloropalladation of 1,6-enynes. AB - A PdCl(2)-catalyzed cis-chloropalladation-cyclization reaction of various 1,6 enyne substrates was developed. This Pd-catalyzed enyne cyclization reaction represents a new route for the synthesis of stereodefined alpha-halomethylene gamma-butyrolactones, lactams, tetrahydrofurans, and cyclopentanes. A mechanism involving a neighboring coordination group is proposed to explain the experiment results. PMID- 15823014 TI - Mechanistic basis of enzyme-targeted drugs. AB - Enzymes offer unique opportunities for drug design that are not available to cell surface receptors, nuclear hormone receptors, ion channels, transporters, and DNA. Here, we review the variety of inhibition mechanisms for enzyme-targeted drugs, and establish an enzyme target database for drugs currently marketed in the United States. From an analysis of the FDA Orange Book, there are 317 marketed drugs that work by inhibiting an enzyme. These drugs inhibit 71 enzymes, including 48 human, 13 bacterial, five viral, four fungal, and one protozoal enzyme. Among the 317 drugs, 65% either undergo reactive chemistry in the active site of the target enzyme or contain a structural motif related to the substrate. Among the 71 enzyme targets, 25 are irreversibly inhibited by drugs, and 19 of the 25 irreversibly inhibited enzymes are covalently modified by the drug. In two additional cases, the drug forms a covalent complex with the substrate, and in three more cases, the drug traps a covalent enzyme-substrate intermediate. Four of the 71 enzymes are inhibited by transition-state analogues. Moreover, advanced methods for determining transition-state structure now offer the opportunity for direct drug design without resorting to expensive random testing campaigns. A full appreciation of enzyme mechanisms sets enzymes apart as a specialized class of targets for highly directed drug design. PMID- 15823015 TI - The so-called Listeria innocua ferritin is a Dps protein. Iron incorporation, detoxification, and DNA protection properties. AB - Listeria innocua Dps (DNA binding protein from starved cells) affords protection to DNA against oxidative damage and can accumulate about 500 iron atoms within its central cavity through a process facilitated by a ferroxidase center. The chemistry of iron binding and oxidation in Listeria Dps (LiDps, formerly described as a ferritin) using H(2)O(2) as oxidant was studied to further define the mechanism of iron deposition inside the protein and the role of LiDps in protecting DNA from oxidative damage. The relatively strong binding of 12 Fe(2+) to the apoprotein (K(D) approximately 0.023 microM) was demonstrated by isothermal titration calorimetry, fluorescence quenching, and pH stat experiments. Hydrogen peroxide was found to be a more efficient oxidant for the protein-bound Fe(2+) than O(2). Iron(II) oxidation by H(2)O(2) occurs with a stoichiometry of 2 Fe(2+)/H(2)O(2) in both the protein-based ferroxidation and subsequent mineralization reactions, indicating complete reduction of H(2)O(2) to H(2)O. Electron paramagnetic resonance (EPR) spin-trapping experiments demonstrated that LiDps attenuates the production of hydroxyl radical by Fenton chemistry. DNA cleavage assays showed that the protein, while not binding to DNA itself, protects it against the deleterious combination of Fe(2+) and H(2)O(2). The overall process of iron deposition and detoxification by LiDps is described by the following equations. For ferroxidation, Fe(2+) + Dps(Z)--> [(Fe(2+)) Dps](Z+1) + H(+) (Fe(2+) binding) and [(Fe(2+))-Dps](Z+1) + Fe(2+) + H(2)O(2) --> [(Fe(3+))(2)(O)(2)-Dps](Z+1) + 2H(+) (Fe(2+) oxidation/hydrolysis). For mineralization, 2Fe(2+) + H(2)O(2) + 2H(2)O --> 2Fe(O)OH((core)) + 4H(+) (Fe(2+) oxidation/hydrolysis). These reactions occur in place of undesirable odd-electron redox processes that produce hydroxyl radical. PMID- 15823016 TI - The unusual intersubunit ferroxidase center of Listeria innocua Dps is required for hydrogen peroxide detoxification but not for iron uptake. A study with site specific mutants. AB - The role of the ferroxidase center in iron uptake and hydrogen peroxide detoxification was investigated in Listeria innocua Dps by substituting the iron ligands His31, His43, and Asp58 with glycine or alanine residues either individually or in combination. The X-ray crystal structures of the variants reveal only small alterations in the ferroxidase center region compared to the native protein. Quenching of the protein fluorescence was exploited to assess stoichiometry and affinity of metal binding. Substitution of either His31 or His43 decreases Fe(II) affinity significantly with respect to wt L. innocua Dps (K approximately 10(5) vs approximately 10(7) M(-)(1)) but does not alter the binding stoichiometry [12 Fe(II)/dodecamer]. In the H31G-H43G and H31G-H43G-D58A variants, binding of Fe(II) does not take place with measurable affinity. Oxidation of protein-bound Fe(II) increases the binding stoichiometry to 24 Fe(III)/dodecamer. However, the extent of fluorescence quenching upon Fe(III) binding decreases, and the end point near 24 Fe(III)/dodecamer becomes less distinct with increase in the number of mutated residues. In the presence of dioxygen, the mutations have little or no effect on the kinetics of iron uptake and in the formation of micelles inside the protein shell. In contrast, in the presence of hydrogen peroxide, with increase in the number of substitutions the rate of iron oxidation and the capacity to inhibit Fenton chemistry, thereby protecting DNA from oxidative damage, appear increasingly compromised, a further indication of the role of ferroxidation in conferring peroxide tolerance to the bacterium. PMID- 15823017 TI - Crystal structures of the PNA-porphyrin complex in the presence and absence of lactose: mapping the conformational changes on lactose binding, interacting surfaces, and supramolecular aggregations. AB - The extraordinary recognition specificity of lectins for carbohydrate ligands appears to be violated as they also bind to porphyrins and other noncarbohydrate ligands. In this study, crystal structures of meso-tetrasulfonatophenylporphyrin (H(2)TPPS) bound to peanut agglutinin (PNA) in the presence and absence of lactose were determined. The binding of H(2)TPPS with PNA involved 11 molecules of H(2)TPPS in different supramolecular stacking arrangements associated with a tetramer of PNA in the crystals of the PNA-H(2)TPPS binary complex as well as the PNA-H(2)TPPS-lactose ternary complex. The ternary complex involved lactose binding only to two subunits of the PNA tetramer, which did not have porphyrin interacting in the vicinity of the carbohydrate-binding site. Comparison of the two structures highlighted the plasticity of the carbohydrate-binding site expressed in terms of the conformational change in lactose binding. The unusual quaternary structure of PNA, which results in exposed protein-protein interaction sites, might be responsible for the porphyrin binding. The association of porphyrin in diverse oligomeric stacking arrangements observed in the PNA H(2)TPPS complex suggested the possibility of protein-porphyrin aggregation under abnormal physiological conditions. The structures described here provide a possible native conformation of the carbohydrate-binding site of PNA in the absence of the ligand, highlight mapping of the unsaturated binding surfaces of PNA using porphyrin interactions, indicate new leads toward possible application of this lectin in photodynamic therapy, and exhibit diverse modes of porphyrin lectin interactions with implications to porphyria, a disease that results from abnormal accumulation of porphyrins. PMID- 15823018 TI - Characterization of a large subunit catalase truncated by proteolytic cleavage. AB - The large subunit catalase HPII from Escherichia coli can be truncated by proteolysis to a structure similar to small subunit catalases. Mass spectrometry analysis indicates that there is some heterogeneity in the precise cleavage sites, but approximately 74 N-terminal residues, 189 C-terminal residues, and a 9 11-residue internal fragment, including residues 298-308, are removed. Crystal structure refinement at 2.8 A reveals that the tertiary and quaternary structure of the native enzyme is retained with only very subtle changes despite the loss of 36% of the sequence. The truncated variant exhibits a 1.8 times faster turnover rate and enhanced sensitivity to high concentrations of H(2)O(2), consistent with easier access of the substrate to the active site. In addition, the truncated variant is more sensitive to inhibition, particularly by reagents such as aminotriazole and azide which are larger than substrate H(2)O(2). The main channel leading to the heme cavity is largely unaffected by the truncation, but the lateral channel is shortened and its entrance widened by removal of the C terminal domain, providing an explanation for easier access to the active site. Opening of the entrance to the lateral channel also opens the putative NADPH binding site, but NADPH binding could not be demonstrated. Despite the lack of bound NADPH, the compound I species of both native and truncated HPII are reduced back to the resting state with compound II being evident in the absorbance spectrum only of the heme b-containing H392A variant. PMID- 15823019 TI - Properties of the human erythrocyte glucose transport protein are determined by cellular context. AB - Human erythrocyte hexose transfer is mediated by the glucose transport protein GLUT1 and is characterized by a complexity that is unexplained by available hypotheses for carrier-mediated sugar transport [Cloherty, E. K., Heard, K. S., and Carruthers, A. (1996) Biochemistry 35, 10411-10421]. The study presented here examines the possibility that the operational properties of GLUT1 are determined by host cell environment. A glucose transport-null strain of Saccharomyces cerevisiae (RE700A) was transfected with the p426 GPD yeast expression vector containing DNA encoding the wild-type human glucose transport protein (GLUT1), mutant GLUT1 (GLUT1(338)(-)(A3)), or carboxy-terminal hemagglutinin-polyHis tagged GLUT1 (GLUT1-HA-H6). GLUT1 and GLUT1-HA-H6 are expressed at the yeast cell membrane and restore 2-deoxy-d-glucose, 3-O-methylglucose, and d-glucose transport capacity to RE700A. GLUT1-HA-H6 confers GLUT1-specific sugar transport characteristics to transfected RE700A, including inhibition by cytochalasin B and high-affinity transport of the nonmetabolized sugar 3-O-methylglucose. GLUT1(338)(-)(A3), a catalytically inactive GLUT1 mutant, is expressed but fails to restore RE700A sugar uptake capacity or growth on glucose. In contrast to transport in human red cells, K(m(app)) for 2-deoxy-d-glucose uptake equals K(i(app)) for 2-deoxy-d-glucose inhibition of 3-O-methylglucose uptake. Unlike transport in human red cells or transport in human embryonic kidney cells transfected with GLUT1-HA-H6, unidirectional sugar uptake in RE700A-GLUT1-HA-H6 is not inhibited by reductant and is not stimulated by intracellular sugar. Net uptake of subsaturating 3-O-methylglucose by RE700A-GLUT1-HA-H6 is a simple, first-order process. These findings support the hypothesis that red cell sugar transport complexity is host cell-specific. PMID- 15823020 TI - Cooperativity during the formation of peptide/MHC class II complexes. AB - To generate an effective immune response, class II major histocompatibility complex molecules (MHCII) must present a diverse array of peptide ligands for recognition by T lymphocytes. Peptide/MHCII complexes are stabilized by hydrophobic anchoring of peptide side chains to pockets in the MHCII protein and the formation of hydrogen bonds to the peptide backbone. Many current models of peptide/MHCII association assume an additive and independent contribution of the interactions between major MHCII pockets and corresponding side chains in the peptide. However, significant conformational rearrangements occur in both the peptide and MHCII during binding. Therefore, we hypothesize that peptide binding to MHCII could be viewed as a folding process in which both molecules cooperate to produce the final conformation. To directly test this hypothesis, we adapt a serial mutagenesis strategy to study cooperativity in the interaction of the human MHCII HLA-DR1 and a peptide derived from influenza hemagglutinin. Substitutions in either the peptide or HLA-DR1 that are predicted to interfere with hydrogen bond formation show cooperative effects on complex stability and affinity. Substitution of a peptide side chain that provides a hydrophobic contact also contributes to the cooperative effect, suggesting a role for all energetic sources in the folding process. We propose that cooperativity throughout the peptide-binding groove reflects the folding of segments of the MHCII molecule into helices around the peptide with a concomitant folding of the peptide into a polyproline helix. The implications of cooperativity for peptide/MHCII structure and epitope selection are discussed. PMID- 15823021 TI - Unfolding and refolding of the glutamine-binding protein from Escherichia coli and its complex with glutamine induced by guanidine hydrochloride. AB - The aim of this work was to study the conformational changes of the Escherichia coli glutamine-binding protein (GlnBP) induced by GdnHCl and the effect of the binding of glutamine (Gln) on these processes. To this end, GdnHCl-induced unfolding of GlnBP alone and its GlnBP-Gln complex was studied by protein intrinsic fluorescence, ANS emission fluorescence, and far- and near-UV circular dichroism spectroscopy. The obtained spectroscopic data were interpreted taking into the account the peculiarities of protein three-dimensional structure. In particular, the fact that formation of a complex of GlnBP and Gln, which essentially changes the global structure of protein, affects only insignificantly the microenvironments of tryptophan residues elucidates the similarity of the emission spectra of GlnBP and the GlnBP-Gln complex, and the existence of quenching groups near tyrosine residues and an effective nonradiative Tyr --> Trp and/or Tyr --> Tyr --> Trp energy transfer provide an explanation for the negligibly small contribution of tyrosine to the bulk fluorescence of the native protein and for its increase in protein unfolding. The use of the parametric presentation of fluorescence data showed that both GlnBP unfolding and GlnBP-Gln unfolding are three-step processes (N --> I(1) --> I(2) --> U), though in the case of the GlnBP-Gln complex these stages essentially overlap. Despite the complex character, GlnBP unfolding is completely reversible. The dramatic shift of the N --> I(1) process to higher GdnHCl concentrations for the GlnBP-Gln complex in comparison with GlnBP was shown. PMID- 15823022 TI - A structural model for the osmosensor, transporter, and osmoregulator ProP of Escherichia coli. AB - Transporter ProP of Escherichia coli, a member of the major facilitator superfamily (MFS), acts as an osmosensor and an osmoregulator in cells and after purification and reconstitution in proteoliposomes. H(+)-osmoprotectant symport via ProP is activated when medium osmolality is elevated with membrane impermeant osmolytes. The three-dimensional structure of ProP was modeled with the crystal structure of MFS member GlpT as a template. This GlpT structure represents the inward (or cytoplasm)-facing conformation predicted by the alternating access model for transport. LacZ-PhoA fusion analysis and site-directed fluorescence labeling substantiated the membrane topology and orientation predicted by this model and most hydropathy analyses. The model predicts the presence of a proton pathway within the N-terminal six-helix bundle of ProP (as opposed to the corresponding pathway found within the C-terminal helix bundle of its paralogue, LacY). Replacement of residues within the N-terminal helix bundle impaired the osmotic activation of ProP, providing the first indication that residues outside the C-terminal domain are involved in osmosensing. Some residues that were accessible from the periplasmic side, as predicted by the structural model, were more susceptible to covalent labeling in permeabilized membrane fractions than in intact bacteria. These residues may be accessible from the cytoplasmic side in structures not represented by our current model, or their limited exposure in vivo may reflect constraints on transporter structure that are related to its osmosensory mechanism. PMID- 15823023 TI - Fidelity of mispair formation and mispair extension is dependent on the interaction between the minor groove of the primer terminus and Arg668 of DNA polymerase I of Escherichia coli. AB - The hydrogen bonding interactions between the Klenow fragment of Escherichia coli DNA polymerase I with the proofreading exonuclease inactivated (KF(-)) and the minor groove of DNA were examined with modified oligodeoxynucleotides in which 3 deazaguanine (3DG) replaced guanine. This substitution would prevent a hydrogen bond from forming between the polymerase and that one site on the DNA. If the hydrogen bonding interaction were important, then we should observe a decrease in the rate of reaction. The steady-state and pre-steady-state kinetics of DNA replication were measured with 10 different oligodeoxynucleotide duplexes in which 3DG was placed at different positions. The largest decrease in the rate of replication was observed when 3DG replaced guanine at the 3'-terminus of the primer. The effect of this substitution on mispair extension and formation was then probed. The G to 3DG substitution at the primer terminus decreased the k(pol) for the extension past G/C, G/A, and G/G base pairs but not the G/T base pair. The G to 3DG substitution at the primer terminus also decreased the formation of correct base pairs as well as incorrect base pairs. However, in all but two mispairs, the effect on correct base pairs was much greater than that of mispairs. These results indicate that the hydrogen bond between Arg668 and the minor groove of the primer terminus is important in the fidelity of both formation and extension of mispairs. These experiments support a mechanism in which Arg668 forms a hydrogen bonding fork between the minor groove of the primer terminus and the ring oxygen of the deoxyribose moiety of the incoming dNTP to align the 3'-hydroxyl group with the alpha-phosphate of the dNTP. This is one mechanism by which the polymerase can use the geometry of the base pairs to modulate the rate of formation and extension of mispairs. PMID- 15823024 TI - Oxidatively induced DNA-protein cross-linking between single-stranded binding protein and oligodeoxynucleotides containing 8-oxo-7,8-dihydro-2'-deoxyguanosine. AB - The formation of covalent cross-links between amino acid side chains and DNA bases in DNA-protein complexes is a significant pathway in oxidative damage to the genome, yet much remains to be learned about their chemical structures and mechanisms of formation. In the present study, DNA-protein cross-links (DPCs) were formed between synthetic oligodeoxynucleotides containing an 8-oxo-7,8 dihydro-2'deoxyguanosine (OG) or an 8-oxo-7,8-dihydro-2'-deoxyadenosine (OA) nucleotide and Escherichia coli singled-stranded binding protein (SSB) under oxidative conditions. Studies with various sequences indicated that DNA homopolymers and those lacking 8-oxopurines were less reactive toward DPC formation. DPCs were formed in the presence of HOCl, peroxynitrite, and the one electron oxidants Na(2)IrCl(6), Na(2)IrBr(6), and Na(3)Fe(CN)(6). Protein-protein cross-linking was also observed, particularly for oxidants of high reduction potential such as Na(2)IrCl(6). The adducted oligodeoxynucleotides were sensitive to hot piperidine treatment leading to strand scission at the site of cross linking. In addition, the covalent cross-links were somewhat heat and acid labile, which may be related to the difficulties encountered in obtaining complete characterization of trypsin digests of the DPCs. However, model reactions involving the single amino acids lysine, arginine, and tyrosine, residues known to be involved in base contacts in the DNA:SSB complex, could be studied, and the adduct formed between N(alpha)-acetyllysine methyl ester and an 18-mer containing OG was tentatively characterized by electrospray ionization mass spectrometry as analogues of spiroiminodihydantoin and guanidinohydantoin. A mechanism involving nucleophilic attack of an amino acid side chain (e.g. the epsilon-amino group of lysine) at C5 of a 2-electron oxidized form of OG is proposed. PMID- 15823025 TI - Sequence of ligand binding and structure change in the diphtheria toxin repressor upon activation by divalent transition metals. AB - The diphtheria toxin repressor (DtxR) is an Fe(II)-activated transcriptional regulator of iron homeostatic and virulence genes in Corynebacterium diphtheriae. DtxR is a two-domain protein that contains two structurally and functionally distinct metal binding sites. Here, we investigate the molecular steps associated with activation by Ni(II)Cl(2) and Cd(II)Cl(2). Equilibrium binding energetics for Ni(II) were obtained from isothermal titration calorimetry, indicating apparent metal dissociation constants of 0.2 and 1.7 microM for two independent sites. The binding isotherms for Ni(II) and Cd(II) exhibited a characteristic exothermic-endothermic pattern that was used to infer the metal binding sequence by comparing the wild-type isotherm with those of several binding site mutants. These data were complemented by measuring the distance between specific backbone amide nitrogens and the first equivalent of metal through heteronuclear NMR relaxation measurements. Previous studies indicated that metal binding affects a disordered to ordered transition in the metal binding domain. The coupling between metal binding and structure change was investigated using near-UV circular dichroism spectroscopy. Together, the data show that the first equivalent of metal is bound by the primary metal binding site. This binding orients the DNA binding helices and begins to fold the N-terminal domain. Subsequent binding at the ancillary site completes the folding of this domain and formation of the dimer interface. This model is used to explain the behavior of several mutants. PMID- 15823026 TI - Amino acid residues involved in substrate recognition of the Escherichia coli Orf135 protein. AB - The Escherichia coli Orf135 protein, a MutT-type enzyme, hydrolyzes mutagenic 2 hydroxy-dATP (2-OH-dATP) and 8-hydroxy-dGTP, in addition to dCTP and 5-methyl dCTP, and its deficiency causes increases in both the spontaneous and H(2)O(2) induced mutation frequencies. To identify the amino acid residues that interact with these nucleotides, the Glu-33, Arg-72, Arg-77, and Asp-118 residues of Orf135, which are candidates for residues interacting with the base, were substituted, and the enzymatic activities of these mutant proteins were examined. The mutant proteins with a substitution at the 33rd, 72nd, and 118th amino acid residues displayed activities affected to various degrees for each substrate, suggesting the involvement of these residues in substrate binding. On the other hand, the mutant protein with a substitution at the 77th Arg residue had activitiy similar to that of the wild-type protein, excluding the possibility that this Arg side chain is involved in base recognition. In addition, the expression of some Orf135 mutants in orf135(-) E. coli reduced the level of formation of rpoB mutants elicited by H(2)O(2). These results reveal the residues involved in the substrate binding of the E. coli Orf135 protein. PMID- 15823027 TI - Solution structure of zinc- and calcium-bound rat S100B as determined by nuclear magnetic resonance spectroscopy. AB - The EF-hand calcium-binding protein S100B also binds one zinc ion per subunit with a relatively high affinity (K(d) approximately 90 nM) [Wilder et al., (2003) Biochemistry 42, 13410-13421]. In this study, the structural characterization of zinc binding to calcium-loaded S100B was examined using high-resolution NMR techniques, including structural characterization of this complex in solution at atomic resolution. As with other S100 protein structures, the quaternary structure of Zn(2+)-Ca(2+)-bound S100B was found to be dimeric with helices H1, H1', H4, and H4' forming an X-type four-helix bundle at the dimer interface. NMR data together with mutational analyses are consistent with Zn(2+) coordination arising from His-15 and His-25 of one S100B subunit and from His-85 and Glu-89 of the other subunit. The addition of Zn(2+) was also found to extend helices H4 and H4' three to four residues similar to what was previously observed with the binding of target proteins to S100B. Furthermore, a kink in helix 4 was observed in Zn(2+)-Ca(2+)-bound S100B that is not in Ca(2+)-bound S100B. These structural changes upon Zn(2+)-binding could explain the 5-fold increase in affinity that Zn(2+)-Ca(2+)-bound S100B has for peptide targets such as the TRTK peptide versus Ca(2+)-bound S100B. There are also changes in the relative positioning of the two EF-hand calcium-binding domains and the respective helices comprising these EF hands. Changes in conformation such as these could contribute to the order of magnitude higher affinity that S100B has for calcium in the presence of Zn(2+). PMID- 15823028 TI - Characterization and structural analyses of nonspecific lipid transfer protein 1 from mung bean. AB - Plant nonspecific lipid transfer proteins (nsLTPs) are thermal stable proteins that are capable of transferring lipid molecules between bilayers in vitro. This family of proteins, abundant in plants, is proposed to be involved in defense, pollination, and germination; the in vivo biological function remains, however, elusive. Here we report the purification and sequencing of an nsLTP1 from mung bean sprouts. We have also determined the solution structure of this nsLTP1, which represents the first 3D structure of the dicotyledonous nsLTP1 family. The global fold of mung bean nsLTP1 is similar to those of the monocotyledonous nsLTP1 structures and consists of four alpha-helices stabilized by four disulfide bonds. There are, however, some notable differences in the C-terminal tails and internal hydrophobic cavities. Circular dichroism and fluorescence spectroscopy were used to compare the thermodynamics and lipid transfer properties of mung bean nsLTP1 with those of rice nsLTP1. Docking of a lipid molecule into the solution structure of mung bean nsLTP1 reveals similar binding cavities and hydrophobic interactions as in rice nsLTP1, consistent with their comparable lipid transfer properties measured experimentally. PMID- 15823029 TI - Identification of the hydrophobic thickness of a membrane protein using fluorescence spectroscopy: studies with the mechanosensitive channel MscL. AB - The hydrophobic thickness of a membrane protein is an important parameter, defining how the protein sits within the hydrocarbon core of the lipid bilayer that surrounds it in a membrane. Here we show that Trp scanning mutagenesis combined with fluorescence spectroscopy can be used to define the hydrophobic thickness of a membrane protein. The mechanosensitive channel of large conductance (MscL) contains two transmembrane alpha-helices, of which the second (TM2) is lipid-exposed. The region of TM2 that spans the hydrocarbon core of the bilayer when MscL is reconstituted into bilayers of dioleoylphosphatidylcholine runs from Leu-69 to Leu-92, giving a hydrophobic thickness of ca. 25 A. The results obtained using Trp scanning mutagenesis were confirmed using Cys residues labeled with the N-methyl-amino-7-nitroben-2-oxa-1,3-diazole [NBD] group; both fluorescence emission maxima and fluorescence lifetimes for the NBD group are sensitive to solvent dielectric constant over the range (2-40) thought to span the lipid headgroup region of a lipid bilayer. Changing phospholipid fatty acyl chain lengths from C14 and C24 results in no significant change for the fluorescence of the interfacial residues, suggesting very efficient hydrophobic matching between the protein and the surrounding lipid bilayer. PMID- 15823030 TI - On the charge regulation of proteins. AB - It is known that the overall charge of a protein can change as the molecule approaches a charged object like another protein or a cell membrane. We have formalized this mechanism using a statistical mechanical framework and show how this rather overlooked interaction increases the attraction between protein molecules. From the theory, we can identify a unique property, the protein charge capacitance, that contains all information needed to describe the charge regulation mechanism. The capacitance can be obtained from experiment or theory and is a function of pH, salt concentration, and the number of titrating residues. For a range of different protein molecules, we calculate the capacitance and demonstrate how it can be used to quantify the charge regulation interaction. With minimal effort, the derived formulas can be used to improve existing models by including a charge regulation term. Good agreement is found between theory, simulations, and experimental data. PMID- 15823031 TI - Crystal structure of Escherichia coli crotonobetainyl-CoA: carnitine CoA transferase (CaiB) and its complexes with CoA and carnitinyl-CoA. AB - L-Carnitine (R-[-]-3-hydroxy-4-trimethylaminobutyrate) is found in both eukaryotic and prokaryotic cells and participates in diverse processes including long-chain fatty-acid transport and osmoprotection. The enzyme crotonobetainyl/gamma-butyrobetainyl-CoA:carnitine CoA-transferase (CaiB; E.C. 2.8.3.-) catalyzes the first step in carnitine metabolism, leading to the final product gamma-butyrobetaine. The crystal structures of Escherichia coli apo-CaiB, as well as its Asp169Ala mutant bound to CoA and to carnitinyl-CoA, have been determined and refined to 1.6, 2.4, and 2.4 A resolution, respectively. CaiB is composed of two identical circular chains that together form an intertwined dimer. Each monomer consists of a large domain, containing a Rossmann fold, and a small domain. The monomer and dimer resemble those of formyl-CoA transferase from Oxalobacter formigenes, as well as E. coli YfdW, a putative type-III CoA transferase of unknown function. The CoA cofactor-binding site is formed at the interface of the large domain of one monomer and the small domain from the second monomer. Most of the protein-CoA interactions are formed with the Rossmann fold domain. While the location of cofactor binding is similar in the three proteins, the specific CoA-protein interactions vary somewhat between CaiB, formyl-CoA transferase, and YfdW. CoA binding results in a change in the relative positions of the large and small domains compared with apo-CaiB. The observed carnitinyl CoA product in crystals of the CaiB Asp169Ala mutant cocrystallized with crotonoyl-CoA and carnitine could result from (i) a catalytic mechanism involving a ternary enzyme-substrate complex, independent of a covalent anhydride intermediate with Asp169, (ii) a spontaneous reaction of the substrates in solution, followed by binding to the enzyme, or (iii) an involvement of another residue substituting functionally for Asp169, such as Glu23. PMID- 15823032 TI - Conjugated bile acid hydrolase is a tetrameric N-terminal thiol hydrolase with specific recognition of its cholyl but not of its tauryl product. AB - Bacterial bile salt hydrolases catalyze the degradation of conjugated bile acids in the mammalian gut. The crystal structures of conjugated bile acid hydrolase (CBAH) from Clostridium perfringens as apoenzyme and in complex with taurodeoxycholate that was hydrolyzed to the reaction products taurine and deoxycholate are described here at 2.1 and 1.7 A resolution, respectively. The crystal structures reveal close relationship between CBAH and penicillin V acylase from Bacillus sphaericus. This similarity together with the N-terminal cysteine classifies CBAH as a member of the N-terminal nucleophile (Ntn) hydrolase superfamily. Both crystal structures show an identical homotetrameric organization with dihedral (D(2) or 222) point group symmetry. The structure analysis of C. perfringens CBAH identifies critical residues in catalysis, substrate recognition, and tetramer formation which may serve in further biochemical characterization of bile acid hydrolases. PMID- 15823033 TI - The N-terminal extracellular domain 23-60 of the calcitonin receptor-like receptor in chimeras with the parathyroid hormone receptor mediates association with receptor activity-modifying protein 1. AB - The calcitonin receptor-like receptor (CLR) requires the associated receptor activity-modifying protein (RAMP)1 to reveal a calcitonin gene-related peptide (CGRP) receptor. Here, the subdomain of the CLR that associates with RAMP1 has been identified in chimeras between the CLR and the parathyroid hormone (PTH) receptor 1 (PTHR). The PTHR alone does not interact with RAMP1. RAMP1 requires the CLR for its transport to the cell surface. Thus, receptor-dependent RAMP1 delivery to the plasma membrane and coimmunoprecipitation from the cell surface were used as measures for receptor/RAMP1 interaction. Several chimeric CLR-PTHR included the N-terminal amino acids 23-60 of the CLR transported RAMP1 to the surface of COS-7 cells much like the intact CLR. Moreover, RAMP1 coimmunoprecipitated with these receptors from the cell surface. A CLR deletion mutant, consisting of the N-terminal extracellular domain, the first transmembrane domain, and the C-terminal intracellular region, revealed the same results. Cyclic AMP was stimulated by CGRP in CLR/RAMP1 expressing cells (58 +/- 19-fold, EC(50) = 0.12 +/- 0.03 nM) and by PTH-related protein in cells expressing the PTHR (50 +/- 10-fold, EC(50) = 0.25 +/- 0.03 nM) or a PTHR with the N-terminal amino acids 23-60 of the CLR (23 +/- 5-fold, EC(50) > 1000 nM). Other chimeric CLR-PTHR were inactive. In conclusion, structural elements in the extreme N-terminus of the CLR between amino acids 23-60 are required and sufficient for CLR/RAMP1 cotransport to the plasma membrane and heterodimerization. PMID- 15823034 TI - Identification of neurite outgrowth active sites on the laminin alpha4 chain G domain. AB - The laminin alpha4 chain is widely distributed in various mesodermal tissues, including the perineurium of peripheral nerves, dorsal root ganglion (DRG), skeletal muscle, and capillaries, and plays important roles in synaptic specialization at the neuromuscular junction and in microvascular formation. The C-terminal globular domain (G domain) of the laminin alpha4 chain was previously found to be critical for heparin binding and cell attachment activity. Here, we focused on neurite outgrowth activity of the laminin alpha4 chain G domain. We found that the recombinant alpha4 chain G domain protein (rec-alpha4G) promoted neurite outgrowth of rat pheochromocytoma PC12 cells. When 114 overlapping synthetic peptides that covered the entire G domain were tested for neurite outgrowth activity, nine peptides were active, but the 105 remaining peptides did not exhibit activity. Three of the nine active peptides, A4G6 (LAIKNDNLVYVY), A4G20 (DVISLYNFKHIY), and A4G107 (VIRDSNVVQLDV), strongly promoted neurite outgrowth of PC12 cells. A4G107 was found to form amyloid-like fibrils in Congo red, X-ray, and electron microscopy analyses. We also synthesized cyclic peptides to evaluate their conformational requirements. Cyclic peptide A4G82X (cyc A4G82X;TLFLAHGRLVFX, where X is norleucine) significantly enhanced neurite outgrowth activity, but the rest of the cyclic peptides eliminated the activity. The A4G82 sequence is located on the loop region, suggesting that the activity of A4G82 is required for a loop conformation. These peptides also exhibited neurite outgrowth activity with dorsal root ganglion (DRG) explants and with DRG cells from E14.5 mouse embryos, indicating that they are active in both neuronal cell lines and native neuronal cells. Taken together, the data suggest that the peptides from the laminin alpha4 chain G domain promote neurite outgrowth activity via a specific conformation. PMID- 15823035 TI - Vmax regulation through domain and subunit changes. The active form of phosphoglycerate dehydrogenase. AB - An active conformation of phosphoglycerate dehydrogenase (PGDH) from Escherichia coli has been obtained using X-ray crystallography. The X-ray crystal structure is used to examine the potential intermediates for V(max) regulation, for the redox reaction, and for cooperative effects of serine binding. The crystal structure at 2.2 A resolution contains bound NAD(+) cofactor, either sulfate or phosphate anions, and alpha-ketoglutarate, a nonphysiological substrate. A PGDH subunit is formed from three distinct domains: regulatory (RBD), substrate (SBD), and nucleotide binding (NBD). The crystal conformation of the homotetramer points to the fact that, in the absence of serine, coordinated movement of the RBD-SBD domains occurs relative to the NBD. The result is a conformational change involving the steric relationships of both the domains and the subunits. Within the active site of each subunit is a bound molecule of alpha-ketoglutarate and the coenzyme, NAD. The catalytic or active site cleft is changed slightly although it is still solvent exposed; therefore, the catalytic reaction probably involves additional conformational changes. By comparing the inhibited with the uninhibited complex, it is possible to describe changes in conformation that are involved in the inhibitory signal transduction of serine. PMID- 15823036 TI - Kindling fluorescent protein from Anemonia sulcata: dark-state structure at 1.38 A resolution. AB - When the nonfluorescent chromoprotein asFP595 from Anemonia sulcata is subjected to sufficiently intense illumination near the absorbance maximum (lambda(abs)(max) = 568 nm), it undergoes a remarkable transition, termed "kindling", to a long-lived fluorescent state (lambda(em)(max) = 595 nm). In the dark recovery phase, the kindled state relaxes thermally on a time scale of seconds or can instantly be reverted upon illumination at 450 nm. The kindling phenomenon is enhanced by the Ala143 --> Gly point mutation, which slows the dark recovery time constant to 100 s at room temperature and increases the fluorescence quantum yield. To investigate the chemical nature of the chromophore and the possible role of chromophore isomerization in the kindling phenomenon, we determined the crystal structure of the "kindling fluorescent protein" asFP595 A143G (KFP) in the dark-adapted state at 1.38 A resolution and 100 K. The chromophore, derived from the Met63-Tyr64-Gly65 tripeptide, closely resembles that of the nonfluorescent chromoprotein Rtms5 in that the configuration is trans about the methylene bridge and there is substantial distortion from planarity. Unlike in Rtms5, in the native protein the polypeptide backbone is cleaved between Cys62 and Met63. The size and shape of the chromophore pocket suggest that the cis isomer of the chromophore could also be accommodated. Within the pocket, partially disordered His197 displays two conformations, which may constitute a binary switch that stabilizes different chromophore configurations. The energy barrier for thermal relaxation was found by Arrhenius plot analysis to be approximately 71 kJ/mol, somewhat higher than the value of approximately 55 kJ/mol observed for cis-trans isomerization of a model chromophore in solution. PMID- 15823037 TI - Synthesis and properties of the chromophore of the asFP595 chromoprotein from Anemonia sulcata. AB - A model compound for the chromophore within the purple nonfluorescent GFP-like chromoprotein asFP595 was synthesized. The postulated structure of the chromophore, 2-acetyl-4-(p-hydroxybenzylidene)-1-methyl-5-imidazolone, was taken from the high-resolution crystal structure analysis of intact asFP595 [Quillin, M. L., Anstrom, D., Shu, X., O'Leary, S., Kallio, K., Lukyanov, K. A., and Remington, S. J. (2005) Kindling Fluorescent Protein from Anemonia sulcata: Dark State Structure at 1.38 A Resolution, Biochemistry 44, 5774-5787]. Erlenmeyer lactonization and oxidation of the methylene group attached to the heteroaromatic moiety with selenium dioxide were used at the key stages of the synthesis. The spectral properties of the model chromophore in solution and their dependence on the pH and polarity of the solvent were investigated. In water, the chromophore was found to exist in two forms, neutral and anionic, with a pK(a) of 7.1. In a dimethylformamide solution, the spectral properties of the anionic form closely match those of the native protein, demonstrating that under these conditions, the compound is an excellent model for the chromophore within native asFP595. PMID- 15823038 TI - Independent and cooperative roles of N-glycans and molecular chaperones in the folding and disulfide bond formation of the low-density lipoprotein (LDL) receptor-related protein. AB - The low-density lipoprotein receptor-related protein (LRP) is a large receptor that contains extensive glycosylation sites and disulfide bonds. Here we analyzed how N-linked glycosylation and molecular chaperones function during LRP folding. Treatment of cells with a glycosylation inhibitor tunicamycin significantly impaired LRP folding, although binding to receptor-associated protein (RAP), a specialized chaperone for LRP, was not affected. The effects of tunicamycin on LRP folding were not due to an inhibition of RAP glycosylation since a mutant RAP that harbors a mutation at its sole glycosylation site was still capable of promoting LRP folding. The roles of N-linked glycosylation and the lectin chaperone, calnexin, in LRP folding were further dissected using LRP minireceptors that carry mutations at individual glycosylation sites. Interestingly, we found that RAP interacts with oxidoreductase ERp57 and mediates its interaction with LRP. Since previous studies have shown that N-glycan-bound calnexin/calreticulin are also capable of recruiting ERp57, our results suggest that N-linked glycosylation and RAP can independently and cooperatively recruit oxidoreductases to facilitate protein folding and proper disulfide bond formation. PMID- 15823039 TI - Nucleolin undergoes partial N- and O-glycosylations in the extranuclear cell compartment. AB - Nucleolin is an ubiquitous, nonhistone nucleolar phosphoprotein involved in fundamental aspects of transcription regulation, cell proliferation, and growth. Nucleolin was primarily found in the nucleus, but it was also proposed as a possible shuttle between the nucleus, cytoplasm, and cell membrane. We report here that part of the extranuclear nucleolin undergoes complex N- and O glycosylations. A band with higher molecular mass (113 kDa) than the 105-kDa classical major nucleolin band was detected on SDS-PAGE gel that cross-reacted with specific anti-nucleolin antibodies and was identified as a nucleolin isoform by mass spectrometry. The presence of N-glycans was first suggested by sensibility of the 113-kDa nucleolin isoform to tunicamycin treatment. Determination of monosaccharide composition by heptafluorobutyrate derivation followed by gas-chromatography mass spectrometry indicated the presence of N- and O-glycans. The structures of N- and O-glycans were first investigated using specificity of binding to lectins. This approach allowed a partial characterization of N-glycan structures and revealed O-glycan structures that could otherwise go unnoticed. Further study of N-glycans by mass spectrometry using direct exoglycosidase treatment on MALDI-TOF target allowed the complete definition of their structures. Finally, the use of peptide mass fingerprinting with sinapinic acid allowed identification of N317 and N492 as the two N glycosylation sites. N317 and N492 belong to RNA-binding domains 1 and 3 of nucleolin, respectively, that suggests a role of glycosylation in regulating the function of the protein. PMID- 15823040 TI - Transport of newly synthesized sterol to the sterol-enriched plasma membrane occurs via nonvesicular equilibration. AB - The mechanism by which newly synthesized sterols are transported from their site of synthesis, the endoplasmic reticulum (ER), to the sterol-enriched plasma membrane (PM) is not fully understood. Studies in mammalian cells suggest that newly synthesized cholesterol is transported to the PM in Golgi-bypassing vesicles and/or via a nonvesicular process. Using the yeast Saccharomyces cerevisiae as a model system, we now rule out an essential role for known vesicular transport pathways in transporting the major yeast sterol, ergosterol, from its site of synthesis to the PM. We use a cyclodextrin-based sterol capture assay to show that transport of newly synthesized ergosterol to the PM is unaltered in cells defective in Sec18p, a protein required for almost all intracellular vesicular trafficking events; we also show that transport is not blocked in cells that are defective in formation of transport vesicles at the ER or in vesicle fusion with the PM. Our data suggest instead that transport occurs by equilibration (t(1/2) approximately 10-15 min) of ER and PM ergosterol pools via a bidirectional, nonvesicular process that is saturated in wild-type exponentially growing yeast. To reconcile an equilibration process with the high ergosterol concentration of the PM relative to ER, we note that a large fraction of PM ergosterol is found condensed with sphingolipids in membrane rafts that coexist with free sterol. We propose that the concentration of free sterol is similar in the PM and ER and that only free (nonraft) sterol molecules have access to a nonvesicular transport pathway that connects the two organelles. This is the first description of biosynthetic sterol transport in yeast. PMID- 15823041 TI - Deimination of histone H2A and H4 at arginine 3 in HL-60 granulocytes. AB - Interplay of various covalent modifications of histone tails has an essential role in regulation of chromatin function. Peptidylarginine deiminase (PADI) 4 deiminates protein arginine to citrulline in a Ca(2+)-dependent manner and is present in the nucleus of granulocyte-differentiated HL-60 cells. When these cells are treated with the calcium ionophore A23187, core histone deimination occurs. To determine the deimination sites of histones, histone species were purified by reverse-phase high-performance liquid chromatography (RP-HPLC) from the cells. Immunoblotting using antimodified citrulline antibody indicated that histones H2A, H3, and H4 but not H2B were deiminated. H2A and H4 were digested with Staphylococcus aureus V8 protease, and the digests were separated by RP HPLC. Immuno dot-blotting and mass spectrometry showed that the deiminated residues were present in H2A (1-56) and H4 (1-52) regions but not in other regions. The H2A peptide (1-56) was digested with alpha-chymotrypsin, and the deiminated peptide was separated from the corresponding nondeiminated peptide by RP-HPLC. The deiminated residue was found to be limited to residues 1-23. Similarly, digestion of the H4 peptide (1-52) with endoproteinase Asp-N and separation of the deiminated peptide from the nondeiminated peptide indicated that the deiminated residue was limited to residues 1-23. Mass spectrometry of lysylendopeptidase digests of the H2A (1-23) and H4 (1-23) peptides showed that deimination occurred at arginine 3 of the N-terminal sequence Ac-SGRGK common to H2A and H4. These results suggest that PADI4 deiminates only a restricted site of target proteins in cells. Deimination of histones is discussed in relation to chromatin structure and function. PMID- 15823042 TI - Kinetic mechanism of channel opening of the GluRDflip AMPA receptor. AB - AMPA-type ionotropic glutamate receptors mediate the majority of fast excitatory neurotransmission in the mammalian central nervous system and are essential for brain functions, such as memory and learning. Dysfunction of these receptors has been implicated in a variety of neurological diseases. Using a laser-pulse photolysis technique, we investigated the channel opening mechanism for GluRD(flip) or GluR4(flip) (i.e., the flip isoform of GluRD), an AMPA receptor subunit. The minimal kinetic mechanism for channel opening is consistent with binding of two glutamate molecules per receptor complex. The GluRD(flip) channel opens with a rate constant of (6.83 +/- 0.74) x 10(4) s(-1) and closes with a rate constant of (3.35 +/- 0.17) x 10(3) s(-1). On the basis of these rate constants, the channel opening probability is calculated to be 0.95 +/- 0.12. Furthermore, the shortest rise time (20-80% of the receptor current response to glutamate) is predicted to be 20 micros, which is approximately 8 times shorter than the previous estimate. These findings suggest that the kinetic property of GluRD(flip) is similar to that of GluR2Q(flip), another fast-activating AMPA receptor subunit. PMID- 15823043 TI - Protein phosphatase regulation of Na+/H+ exchanger isoform I. AB - We investigated regulation of Na(+)/H(+) exchanger isoform 1 (NHE1) by dephosphorylation. Treatment of primary cultures of cardiomyocytes with the phosphatase inhibitor okadaic acid increased the rate of recovery from an acid load, suggesting that the okadaic acid sensitive PP1 may be involved in NHE1 regulation in vivo. We examined the ability of purified protein phosphatases PP1, PP2A, and PP2B to dephosphorylate the regulatory cytoplasmic tail. NHE1 was completely dephosphorylated by PP1, poorly dephosphorylated by PP2A, and not dephosphorylated by PP2B. Examination of NHE1 binding to PP1 or PP2B revealed that an association occurs between NHE1 and PP1 both in vitro and in vivo, but NHE1 did not associate with full-length PP2B. We expressed PP1 or inhibitor 2, a specific PP1 inhibitor, in cell lines to examine the effect of PP1 on NHE1 activity in vivo. Overexpression of PP1 causes a decrease in NHE1 activity but does not affect stimulation by thrombin. Cell lines expressing the specific PP1 inhibitor, inhibitor 2, had elevated proton efflux rates and could not be further stimulated by the Na(+)/H(+) exchanger agonist thrombin. The results suggest that PP1 is an important regulatory phosphatase of NHE1, that it can bind to and dephosphorylate the protein, and that it regulates NHE1 activity in vivo. PMID- 15823044 TI - The role of the N-terminal heptad repeat of HIV-1 in the actual lipid mixing step as revealed by its substitution with distant coiled coils. AB - The gp41 glycoprotein of HIV-1 is considered to be responsible for the actual fusion process between the virus and the host membranes. According to a prevailing model, gp41 trimer organization, directed by the N-terminal coiled coil region (NHR), is essential for steps involved in the actual merging of viral and cellular membranes. This study addresses a major question: Is the specific sequence of the NHR obligatory for the fusion process, or can it be replaced by distant coiled coils that form different oligomeric states in solution? For this purpose we synthesized three known GCN4 coiled-coil mutants that oligomerize in solution into either dimers, trimers, or tetramers. These peptides were chemically ligated to the fusion peptide thereby creating three chimera peptides with different oligomeric tendencies in solution. These peptides were investigated, together with the 70-mer wild-type peptide (N70), regarding their structure in solution and membrane by using circular dichroism (CD) and FTIR spectroscopies, their ability to induce vesicle fusion, and their ability to bind phospholipid membranes by using surface plasmon resonance (SPR). Our results suggest that local assembly of fusion peptides, facilitated by coiled-coil oligomers, increases lipid mixing ability, probably by facilitating stronger binding of the fusion peptides to the opposing membrane as revealed by SPR. However, N70 is significantly more active than the other chimeras. Overall, the data indicate a correlation between the distinct conformation of N70 in solution and in membranes and its enhanced lipid mixing relative to the GCN4 chimeras. PMID- 15823045 TI - Triggers of full-length tau aggregation: a role for partially folded intermediates. AB - Alzheimer's disease is characterized in part by the accumulation of full-length tau proteins into intracellular filamentous inclusions. To clarify the events that trigger lesion formation, the aggregation of recombinant full-length four repeat tau (htau40) was examined in vitro under near-physiological conditions using transmission electron microscopy and spectroscopy methods. In the absence of exogenous inducers, tau protein behaved as an assembly-incompetent monomer with little tertiary structure. The addition of anionic inducers led to fibrillization with nucleation-dependent kinetics. On the basis of circular dichroism spectroscopy and reactivity with thioflavin S and 8-anilino-1 naphthalenesulfonic acid fluorescent probes, the inducer stabilized a monomeric species with the folding characteristics of a premolten globule state. Planar aromatic dyes capable of binding the intermediate state with high affinity were also capable of triggering fibrillization in the absence of other inducers. Dye mediated aggregation was characterized by concentration-dependent decreases in lag time, indicating increased nucleation rates, and submicromolar critical concentrations, indicating a final equilibrium that favored the filamentous state. The data suggest that the rate-limiting barrier for filament formation from full-length tau is conformational and that the aggregation reaction is triggered by environmental conditions that stabilize assembly-competent conformations. PMID- 15823046 TI - Heterogeneity in the binding of lipid molecules to the surface of a membrane protein: hot spots for anionic lipids on the mechanosensitive channel of large conductance MscL and effects on conformation. AB - We have introduced single Trp residues into the mechanosensitive channel of large conductance (MscL) from Mycobacterium tuberculosis and used fluorescence quenching by brominated phospholipids to detect the presence of a binding site of high affinity for anionic phospholipids. A cluster of three positively charged residues, Arg-98, Lys-99, and Lys-100, is located on the cytoplasmic side of MscL, in a position where they could interact with the headgroup of an anionic phospholipid. Single mutations of these charged residues in the Trp-containing mutant F80W results in a decreased affinity for phosphatidic acid. Single mutations of the charged residues also result in a significant shift in the fluorescence emission spectrum in dioleoylphosphatidylcholine [di(C18:1)PC] but smaller shifts in dioleoylphosphatidic acid [di(C18:1)PA], suggesting that single mutations result in a conformational change for the protein that is reversed by interaction with anionic phospholipids. This is consistent with the observation that single mutations of the charged residues do not result in a gain of function phenotype. In contrast, simultaneous mutation of all three charged residues results in a gain of function phenotype, and a shift in fluorescence emission spectrum in di(C18:1)PC not reversed in di(C18:1)PA. The gain of function mutant F80W:V21K also shows a shifted fluorescence emission spectrum in both di(C18:1)PC and di(C18:1)PA and binds di(C18:1)PC and di(C18:1)PA with equal affinity, suggesting that the conformational change caused by the V21K mutation results in a breakup of the cluster of three positive charges. Experiments with the Trp mutants L69W and Y87W allow us to measure lipid binding constants on the periplasmic and cytoplasmic sides of the membrane, respectively. On both sides of the membrane the affinity for di(C18:1)PC is equal to that for dioleoylphosphatidylethanolamine. On the periplasmic side of the membrane, there is no selectivity for anionic phospholipids. In contrast, quenching data for Y87W provides evidence for the existence of two lipid binding sites on the cytoplasmic side of the membrane close to the Trp residue at position 87, with binding to one of these sites showing a marked preference for anionic lipid over zwitterionic lipid, presumably involving the charged cluster Arg-98, Lys-99, and Lys-100. PMID- 15823047 TI - Stable recognition of TA interruptions by triplex forming oligonucleotides containing a novel nucleoside. AB - We have prepared the 2'-aminoethoxy derivative of the S nucleoside ((2AE)S) and incorporated it into triplex-forming oligonucleotides for recognition of TA interruptions within a target oligopurine tract. Fluorescence melting, UV melting, and DNase I footprinting experiments show that (2AE)S has greater affinity than G or S for a single TA interruption. Stable triplexes are formed at pH 6.0 at an 18-mer target site containing two TA interruptions, even though this contains eight C(+).GC triplets. Although (2AE)S and S produce stable triplexes at TA interruptions, they also interact with other base pairs, in particular, CG, although the selectivity for TA improves with increased pH.( 2AE)S is the best nucleoside described so far for recognition of TA within a triple-helix target. PMID- 15823048 TI - The inhibitory effect of HMGB-1 protein on the repair of cisplatin-damaged DNA is accomplished through the acidic domain. AB - The well established inhibitory effect of HMGB-1 on repair of cisplatin-damaged DNA has been studied with two modified forms of the protein, shown to bind platinated DNA with higher affinity than the original protein: in vivo acetylated HMGB-1 and HMGB-1 lacking its C-terminal domain. The native and the modified proteins were assayed for their effects on adduct removal by using cell-free extract capable of repairing cisplatinated DNA in vitro. The inhibition observed with the native HMGB-1 was reduced in the presence of acetylated HMGB-1 and completely abolished when the assay was carried out with the truncated protein. When the repair assay was performed in the presence of a synthetic polypeptide identical to the C-terminal tail, either alone or together with the truncated protein, the inhibitory effect was partially recovered in a concentration dependent manner. These findings strongly suggest that the HMGB-1-induced inhibition of cisplatin-DNA adduct repair is accomplished through the acidic domain. The results obtained are discussed in terms of the repair events that may occur in the presence of HMGB-1 protein. PMID- 15823049 TI - Increasing the reactivity of an artificial dithiol-disulfide pair through modification of the electrostatic milieu. AB - The thiol-disulfide exchange reaction plays a central role in the formation of disulfide bonds in newly synthesized proteins and is involved in many aspects of cellular metabolism. Because the thiolate form of the cysteine residue is the key reactive species, its electrostatic milieu is thought to play a key role in determining the rates of thiol disulfide exchange reactions. While modest reactivity effects have previously been seen in peptide model studies, here, we show that introduction of positive charges can have dramatic effects on disulfide bond formation on a structurally restricted surface. We have studied properties of vicinal cysteine residues in proteins using a model system based on redox sensitive yellow fluorescent protein (rxYFP). In this system, the formation of a disulfide bond between two cysteines Cys149 and Cys202 is accompanied by a 2.2 fold decrease in fluorescence. Introduction of positively charged amino acids in the proximity of the two cysteines resulted in an up to 13-fold increase in reactivity toward glutathione disulfide. Determination of the individual pK(a) values of the cysteines showed that the observed increase in reactivity was caused by a decrease in the pK(a) value of Cys149, as well as favorable electrostatic interactions with the negatively charged reagents. The results presented here show that the electrostatic milieu of cysteine thiols in proteins can have substantial effects on the rates of the thiol-disulfide exchange reactions. PMID- 15823050 TI - Dismutase activity of ADP-L-glycero-D-manno-heptose 6-epimerase: evidence for a direct oxidation/reduction mechanism. AB - The first positive evidence for the utilization of a direct C-6' ' oxidation/reduction mechanism by ADP-l-glycero-d-manno-heptose 6-epimerase is reported here. The epimerase (HldD or AGME, formerly RfaD) operates in the biosynthetic pathway of l-glycero-d-manno-heptose, which is a conserved sugar in the core region of lipopolysaccharide (LPS) of Gram-negative bacteria. The stereochemical inversion catalyzed by the epimerase is interesting as it occurs at an "unactivated" stereocenter that lacks an acidic C-H bond, and therefore, a direct deprotonation/reprotonation mechanism cannot be employed. Instead, the epimerase employs a transient oxidation strategy involving a tightly bound NADP(+) cofactor. A recent study ruled out mechanisms involving transient oxidation at C-4' ' and C-7' ' and supported a mechanism that involves an initial oxidation directly at the C-6' ' position to generate a 6' '-keto intermediate (Read, J. A., Ahmed, R. A., Morrison, J. P., Coleman, W. G., Jr., Tanner, M. E. (2004) J. Am. Chem. Soc. 126, 8878-8879). A subsequent nonstereospecific reduction of the ketone intermediate can generate either epimer of the ADP heptose. In this work, an intermediate analogue containing an aldehyde functionality at C-6' ', ADP-beta-d-manno-hexodialdose, is prepared in order to probe the ability of the enzyme to catalyze redox chemistry at this position. It is found that incubation of the aldehyde with a catalytic amount of the epimerase leads to a dismutation process in which one-half of the material is oxidized to ADP-beta-d-mannuronic acid and the other half is reduced to ADP-beta-d-mannose. Transient reduction of the enzyme-bound NADP(+) was monitored by UV spectroscopy and implicates the cofactor's involvement during catalysis. PMID- 15823051 TI - Heterologously expressed bacterial and human multidrug resistance proteins confer cadmium resistance to Escherichia coli. AB - The human MDR1 gene is induced by cadmium exposure although no resistance to this metal is observed in human cells overexpressing hMDR1. To access the role of MDR proteins in cadmium resistance, human MDR1, Lactococcus lactis lmrA, and Oenococcus oeni omrA were expressed in an Escherichia coli tolC mutant strain which proved to be hypersensitive to cadmium. Both the human and bacterial MDR genes conferred cadmium resistance to E. coli up to 0.4 mM concentration. Protection was abolished by 100 microM verapamil. Quantification of intracellular cadmium concentration by atomic absorption spectrometry showed a reduced cadmium accumulation in cells expressing the MDR genes. Inside-out membrane vesicles of L. lactis overexpressing lmrA displayed an ATP-dependent (109)Cd(2+) uptake that was stimulated by glutathione. An evolutionary model is discussed in which MDR proteins have evolved independently from an ancestor protein displaying both organic xenobiotic- and divalent metal-extrusion abilities. PMID- 15823052 TI - Attempts to delineate the relative contributions of changes in hydrophobicity and packing to changes in stability of ribonuclease S mutants. AB - While the hydrophobic driving force is thought to be a major contributor to protein stability, it is difficult to experimentally dissect out its contribution to the overall free energy of folding. We have made large to small substitutions of buried hydrophobic residues at positions 8 and 13 in the peptide/protein complex, RNase-S, and have characterized the structures by X-ray crystallography. The thermodynamics of association of these mutant S peptides with S protein was measured in the presence of different concentrations of methanol and ethanol. The reduction in the strength of the hydrophobic driving force in the presence of these organic solvents was estimated from surface-tension data as well as from the dependence of the DeltaC(p) of protein/peptide binding on the alcohol concentration. The data indicated a decrease in the strength of the hydrophobic driving force of about 30-40% over a 0-30% range of the alcohol concentration. We observe that large to small substitutions destabilize the protein. However, the amount of destabilization, relative to the wild type, is independent of the alcohol concentration over the range of alcohol concentrations studied. The data clearly indicate that decreased stability of the mutants is primarily due to the loss of packing interactions rather than a reduced hydrophobic driving force and suggest a value of the hydrophobic driving force of less than 18 cal mol(-)(1) A(2). PMID- 15823053 TI - Herbicide-induced changes in charge recombination and redox potential of Q(A) in the T4 mutant of Blastochloris viridis. AB - To gain new insights into the function of photosystem II (PSII) herbicides DCMU (a urea herbicide) and bromoxynil (a phenolic herbicide), we have studied their effects in a better understood system, the bacterial photosynthetic reaction center of the terbutryn-resistant mutant T4 of Blastochloris (Bl.) viridis. This mutant is uniquely sensitive to these herbicides. We have used redox potentiometry and time-resolved absorption spectroscopy in the nanosecond and microsecond time scale. At room temperature the P(+)(*)Q(A)(-)(*) charge recombination in the presence of bromoxynil was faster than in the presence of DCMU. Two phases of P(+)(*)Q(A)(-)(*) recombination were observed. In accordance with the literature, the two phases were attributed to two different populations of reaction centers. Although the herbicides did induce small differences in the activation barriers of the charge recombination reactions, these did not explain the large herbicide-induced differences in the kinetics at ambient temperature. Instead, these were attributed to a change in the relative amplitude of the phases, with the fast:slow ratio being approximately 3:1 with bromoxynil and approximately 1:2 with DCMU at 300 K. Redox titrations of Q(A) were performed with and without herbicides at pH 6.5. The E(m) was shifted by approximately -75 mV by bromoxynil and by approximately +55 mV by DCMU. As the titrations were done over a time range that is assumed to be much longer than that for the transition between the two different populations, the potentials measured are considered to be a weighted average of two potentials for Q(A). The influence of the herbicides can thus be considered to be on the equilibrium of the two reaction center forms. This may also be the case in photosystem II. PMID- 15823054 TI - Product release is rate-limiting in the activation of the prodrug 5 fluorocytosine by yeast cytosine deaminase. AB - Yeast cytosine deaminase (yCD), a zinc metalloenzyme, catalyzes the hydrolytic deamination of cytosine to uracil. The enzyme is of great biomedical interest because it also catalyzes the deamination of the prodrug 5-fluorocytosine (5FC) to form the anticancer drug 5-fluorouracil (5FU). yCD/5FC is one of the most widely used enzyme/prodrug combinations for gene-directed enzyme prodrug therapy for the treatment of cancers. A pH indicator assay has been developed for the measurement of the steady-state kinetic parameters for the deamination reaction. Transient kinetic studies have shown that the product release is a rate-limiting step in the activation of the prodrug 5FC by yCD. The rate constant of the chemical step for the forward reaction (250 s(-)(1)) is approximately 8 times that of the product release (31 s(-)(1)) and approximately 15 times k(cat) (17 s( )(1)). The transient kinetic results are consistent with those of the steady state kinetic analysis in the sense that the k(cat) and K(m) values calculated from the rate constants determined by the transient kinetic analysis are in close agreement with those measured by the steady-state kinetic analysis. NMR experiments have demonstrated that free 5FU is in slow exchange with its complex with yCD but has a low affinity for yCD. The transient kinetic and NMR results together suggest that the release of 5FU is rate-limiting in the activation of the prodrug 5FC by yCD and may involve multiple steps. PMID- 15823056 TI - Marked dystrophy of fingernail plates. Topical and systemic antifungal agents offer no relief. PMID- 15823057 TI - Bereavement in late life: risk factors for complicated bereavement. PMID- 15823058 TI - Aging with mental retardation: increasing population of older adults with MR require health interventions and prevention strategies. AB - The second half of the 20th century vastly increased the life span of those with mental retardation (MR). Today the average life expectancy of older adults with MR is 66.1 years and growing. As these individuals age, they present increasing challenges to the clinician. Increased rates of hearing and visual impairments, obesity, and osteoporosis as well as high rates of dementia with associated psychiatric problems make care for the aging individual with MR complex. Primary care providers need to be aware that elders with MR will comprise an increasing part of their practice, and that they present with many chronic conditions. PMID- 15823059 TI - Herb-drug interactions: interactions between Ginkgo biloba and prescription medications. AB - Geriatric patients typically present with one chronic disease per decade over age 50. Each chronic disease typically requires long-term drug therapy, meaning most older patients require several drugs to control their conditions and/or maintain their health. Simultaneously, the use of complementary and alternative medications (CAM) has increased in the United States over the last 20 years, reaching 36% in 2002; herbal medicine use accounts for approximately 22% of all CAM use. Older adults often add herbal medicines to the medications prescribed by their physicians, yet do not always inform their physicians. The drug metabolizing enzyme systems process all compounds foreign to the body, including prescription drugs and herbal medications. Therefore, use of both medicinals simultaneously has a potential for interactions of an adverse nature. This review, which will discuss ginkgo biloba, is the first of a continuing series covering the documented interactions between herbal medicines with proven efficacy and prescription drugs. PMID- 15823060 TI - Preventive services: counseling for healthy lifestyles. PMID- 15823061 TI - Neurological outcome of conservative versus endovascular treatment of patients with asymptomatic high-grade carotid artery stenosis: a propensity score-adjusted analysis. AB - PURPOSE: To report a propensity score-adjusted analysis of the long-term risks for stroke after carotid artery stenting (CAS) compared to medical therapy in patients with asymptomatic high-grade carotid artery stenosis. METHODS: A total of 946 consecutive patients (605 men; median age 73 years) with asymptomatic high grade carotid artery stenoses (> or =70%) identified in a single center registry were treated either medically (n=525) or with CAS (n=421). A propensity score adjusted analysis was performed to test the hypothesis that long-term neurological outcome might be better after CAS than after medical treatment, depending on the baseline degree of carotid stenosis and the patient's medical status. Baseline degree of stenosis was classified as 70% to 79% (n=307), 80% to 89% (n=366), and 90% to 99% (n=272) by duplex ultrasound. Surgical risk was estimated by the American Society of Anesthesiologists (ASA) score (I to IV). RESULTS: Stroke-free survival rates at 1, 3, and 5 years were 97%, 93%, and 89% after conservative treatment versus 94%, 93%, and 91% after CAS (p=0.56), respectively. Compared to conservatively treated patients with 70% to 79% stenoses, the adjusted hazard ratios for stroke were 2.36 (p=0.044) for conservatively treated patients with 80% to 89% and 3.17 (p=0.026) for those with 90% to 99% stenoses. For CAS patients with 70% to 79%, 80% to 89%, and 90% to 99% stenoses, the adjusted hazard ratios for stroke were 1.32 (p=0.63), 0.91 (p=0.84), and 0.98 (p=0.98) irrespective of the ASA score, the propensity to undergo CAS, and other potential confounders. Thus, the risk of stroke increased in parallel with the degree of stenosis in conservatively treated patients, but remained unchanged in patients undergoing CAS. CONCLUSIONS: Patients with asymptomatic severe carotid narrowing (> or =80%) might benefit from CAS with respect to stroke-free survival. Randomized controlled trials are needed to confirm these findings. PMID- 15823062 TI - Proximal endovascular flow blockage for cerebral protection during carotid artery stenting: results from a prospective multicenter registry. AB - PURPOSE: To evaluate the feasibility of cerebral protection during carotid artery stenting (CAS) using the Mo.Ma device, which prevents cerebral embolization by proximal endovascular blockage of blood flow in the internal and external carotid arteries. METHODS: In 14 European centers, 157 patients (121 men; mean age 68.0+/ 8.3 years) were enrolled in a prospective registry between March 2002 and March 2003. Eligible patients had a symptomatic (>50%) or asymptomatic (>70%) stenosis of the internal carotid artery suitable for carotid stenting. Protected carotid stenting was performed with the Mo.Ma system, which occludes both the common and external carotid arteries via 2 independently inflatable compliant low-pressure balloons before any device is advanced across the lesion. Blood is aspirated through the catheter intermittently or at the end of the procedure. RESULTS: The device was successfully positioned and stents were implanted in all cases. Diameter stenosis was reduced from 84.1%+/-7.8% to 6.7%+/-5.1%. The mean duration of flow blockage was 7.6+/-5.9 minutes. In 12 (7.6%) patients, transient intolerance to flow blockade was observed, but the procedures were completed successfully. In 124 (79.6%) cases, there was macroscopic evidence of debris after filtering the aspirated blood. In-hospital complications included 4 (2.5%) minor strokes, 8 (5.1%) transient ischemic attacks, no deaths, and no major strokes, resulting in a 2.5% death/stroke rate at discharge. At 30-day follow-up, there was 1 cardiac death, 1 major stroke, and 3 minor strokes, for an overall 5.7% 30-day death/stroke rate and a 30-day major stroke and death rate of 1.3%. CONCLUSIONS: Cerebral protection with proximal endovascular blood flow blockage during CAS is feasible, with a high procedural success rate. PMID- 15823063 TI - C-type natriuretic peptide inhibits constrictive remodeling without compromising re-endothelialization in balloon-dilated renal arteries. AB - PURPOSE: To investigate the long-term effect of local, liposome-mediated gene transfer of C-type natriuretic peptide (CNP) plasmid versus CNP protein on restenosis in porcine renal arteries following balloon angioplasty. METHODS: The renal arteries of 15 pigs were dilated and the adventitia at the site of balloon injury injected with CNP protein, pCR3.1 plasmid encoding CNP, or the beta galactosidase gene (control) via a needle injection catheter. Five animals receiving the CNP and control genes in dilated arteries were sacrificed after 3 weeks to analyze re-endothelialization, proliferation, and early CNP expression. Ten animals designated for the long-term experiments (3 months) were treated with the CNP gene versus CNP protein (n=3), the CNP gene versus the control gene (n=3), and the CNP protein versus the control gene (n=3). One animal served as a dilated non-treated control. Transfection and expression of CNP and beta galactosidase were measured by polymerase chain reaction (PCR) and reverse transcription PCR. Renal arterial lumen narrowing was measured with angiography and histology. Endothelialization was assessed using Evans blue stain; vWF, CD31, factor VIII, and Ki67 were markers for immunohistochemical analysis. RESULTS: An intact endothelial layer was seen at 3 weeks following angioplasty in all transfected arteries. Three months following treatment, computer-assisted morphometric analysis revealed significant enlargement of the arterial cross sectional areas in CNP plasmid- treated vessels compared to dilated but untreated arteries (CNP plasmid +34.8%+/-13.9% versus CNP protein -1.75%+/-19.9% versus beta-galactosidase -47.0%+/-13.9%, p<0.01). Angiographic analysis showed significant enlargement of the arterial diameter compared to dilated, untreated arteries (CNP plasmid +20.8%+/-6.8% versus CNP protein +5.7%+/-6.0% versus beta galactosidase -24.5%+/-10.2%, p<0.01). CONCLUSIONS: Local application of CNP plasmid proved superior to CNP protein in producing rapid re-endothelialization and significantly enlarging the renal arterial lumen following dilation. PMID- 15823064 TI - Does sac size matter? Findings based on surgical exploration of excluded abdominal aortic aneurysms. AB - PURPOSE: To prospectively examine the outcomes of excluded abdominal aortic aneurysms (AAA) that continue to expand without evidence of endoleak. METHODS: From 1984 to 1998, 1218 patients underwent operative retroperitoneal exclusion of AAA and aortoiliac reconstructions. During the procedure, the aneurysm sac was ligated proximally, as well as distally, which created an ideal in-vivo model of excluded AAA sacs with or without endoleaks. From January 2002 to June 2003, 15 of these patients were identified as having an increase in AAA sac size with or without an endoleak on duplex ultrasonography. These patients were prospectively evaluated by computed tomography and diagnostic arteriography. Patients with a demonstrable endoleak underwent embolization, and the remainder underwent open surgical exploration. RESULTS: Eight patients had arteriographically demonstrated endoleaks that were treated with coil embolization. The remaining 7 patients (6 men; mean age 76 years, range 68-81) without a demonstrable endoleak underwent elective surgical exploration and sac endoaneurysmorrhaphy. The mean time interval between the original surgery and aneurysm sac exploration was 76 months (range 52-92); during this time, the mean aneurysm sac size increased by 2.7 cm (range 1.3-5.2). The mean sac pressure was 53 mmHg, and the sac walls were noticeably thickened, with markedly dilated vasa vasorum. The sac contained yellow, fibrinous material with clear serous fluid (5 patients without any evidence of retrograde flow) or liquefied thrombus with serosanguinous fluid (2 patients with retrograde flow from lumbar arteries). No AAA sacs were pulsatile. CONCLUSIONS: Continued expansion of excluded AAA sacs can occur from causes other than a missed endoleak. Exudation of fluid from thickened sac wall and vasa vasorum, as well as local enzymatic activity, might lead to the formation of a sac hygroma. Furthermore, these findings raise questions as to the need for surgical exploration of all patients with an enlarging AAA sac in the setting of low sac pressures and no definable endoleak. PMID- 15823065 TI - Preserved pelvic circulation after stent-graft treatment of complex aortoiliac artery aneurysms: a new approach. AB - PURPOSE: To describe an endovascular technique that allows stent-graft treatment of aortoiliac aneurysmal disease affecting both common iliac arteries (CIA), with maintenance of pelvic circulation on one side. TECHNIQUE: For patients with aortoiliac aneurysms, both common femoral arteries (CFA) were surgically exposed. One internal iliac artery (IIA) was initially embolized with coils. A bifurcated stent-graft main body was deployed with the proximal end just below the renal arteries. On the ipsilateral side, the stent-graft limb was extended 3 cm beyond the orifice of the embolized IIA into the external iliac artery (EIA) using stent graft limb extenders. On the contralateral side, the stent-graft limb was deployed so that the distal end was 10 to 15 mm proximal to the patent IIA orifice. Via a left brachial artery access, the IIA was catheterized, and stent grafts were deployed from the distal end of the contralateral AAA stent-graft limb into the IIA. A femorofemoral crossover graft provided circulation to the leg ipsilateral to the IIA stent-graft, and the EIA on the same side was ligated. The technique can also be modified to treat isolated bilateral CIA aneurysms. CONCLUSIONS: By extending the distal aspect of the stent-graft into an IIA, bilateral CIA aneurysms can be excluded while preserving pelvic circulation on one side. PMID- 15823066 TI - Three-dimensional teleradiology for surveillance following endovascular aortic aneurysm repair: a feasibility study. AB - PURPOSE: To study the feasibility of 3-dimensional (3D) teleradiology in surveillance of patients treated with stent-grafts for abdominal aortic aneurysm (AAA). METHODS: Between April 2002 and November 2003, 8 AAA patients (7 men; median age 73 years, range 62-84) with stent-grafts had follow-up computed tomograms (CT) performed at their local hospital and transmitted without loss across a broadband connection to the university hospital. On both monitors, the radiologists were presented with the complete CT axial dataset, sagittal and coronal reformatted slices, and a 3D volume-rendered reconstruction. The two radiologists were then able to simultaneously perform measurements and real-time manipulations of the axial and 3D pictures, which were discussed over the telephone or using a videoconferencing unit. Patient satisfaction, the radiologists' evaluation of the method, and the potential cost savings were explored. RESULTS: Twelve follow-up CT scans were performed on the 8 patients. The time for transmission over the teleradiological network averaged 5 minutes, and the evaluation required 15 minutes at the university hospital. The overall technical quality of the images was rated as good by the university radiologist. In 11 studies, the stent-grafts were satisfactory, but a type III endoleak was detected in one 5.5-year-old stent-graft. Neither radiologist had a problem identifying the endoleak. Patients had confidence that the examination at the local hospital was of good quality; they all felt that they received good care and were pleased with avoiding travel to the university hospital. From the economic analysis, an annual savings of 40,000 Euros (USD 52,304) was projected, mostly due to avoiding hospital stays and outpatient consultations at the university hospital. CONCLUSIONS: The experiences from this study are encouraging, but a larger series will be necessary for a thorough evaluation of 3D teleradiology as a surveillance method for aortic stent-graft patients. PMID- 15823067 TI - Telementoring during endovascular treatment of abdominal aortic aneurysms: a prospective study. AB - PURPOSE: To explore the use of telementoring for distant teaching and training in endovascular aortic aneurysm repair (EVAR). METHODS: According to a prospectively designed study protocol, 48 patients underwent EVAR: the first 12 patients (group A) were treated at a secondary care center by an experienced interventionist, who was training the local team; a further 12 patients (group B) were operated by the local team at their secondary center with telementoring by the experienced operator from an adjacent suite; and the last 24 patients (group C) were operated by the local team with remote telementoring support from the experienced interventionist at a tertiary care center. Telementoring was performed using 3 video sources; images were transmitted using 4 ISDN lines. EVAR was performed using intravascular ultrasound and simultaneous fluoroscopy to obtain road mapping of the abdominal aorta and its branches, as well as for identifying the origins of the renal arteries, assessing the aortic neck, and monitoring the attachment of the stent-graft proximally and distally. RESULTS: Average duration of telementoring was 2.1 hours during the first 12 patients (group B) and 1.2 hours for the remaining 24 patients (group C). There was no difference in procedural duration (127+/-59 minutes in group A, 120+/-4 minutes in group B, and 119+/-39 minutes in group C; p=0.94) or the mean time spent in the ICU (26+/-15 hours in group A, 22+/-2 hours in group B, and 22+/-11 hours for group C; p=0.95). The length of hospital stay (11+/-4 days in group A, 9+/-4 days in group B, and 7+/-1 days in group C; p=0.002) was significantly different only for group C versus A (p=0.002). Only 1 (8.3%) patient (in group A: EVAR performed by the experienced operator) required conversion to open surgery because of iliac artery rupture. This was the only conversion (and the only death) in the entire study group (1/12 in group A versus 0/36 in groups B + C, p=0.31). CONCLUSIONS: Telementoring for EVAR is feasible and shows promising results. It may serve as a model for development of similar projects for teaching other invasive procedures in cardiovascular medicine. PMID- 15823068 TI - AngioJet rheolytic thrombectomy versus local intrapulmonary thrombolysis in massive pulmonary embolism: a retrospective data analysis. AB - PURPOSE: To compare the efficacy of full-dose local intrapulmonary thrombolysis (LIT) versus AngioJet rheolytic thrombectomy (ART) in the treatment of massive pulmonary embolism. METHODS: A retrospective review was conducted of 8 consecutive patients (5 women; mean age 66.0+/-5.9 years, range 56-74) who underwent LIT with high-dose intrapulmonary urokinase (4400 IU/kg over 10 minutes followed by a 2000-IU/kg/h infusion) and a subsequent 6 consecutive patients (4 men; mean age of 59.2+/-17.0 years, range 26-69) who underwent ART plus adjunctive low-dose urokinase infusions (100,000 IU) until hemodynamic recovery was achieved. Pre and postprocedural Miller scores were calculated, and relative Miller score improvement, total urokinase doses, and duration of therapy were compared. RESULTS: Hemodynamic stability was restored in all 8 LIT patients and in 5 (83%) of the 6 ART patients; 1 (16.7%) patient died during the ART procedure due to recurrent MPE. In the LIT group, the mean Miller score prior to intervention was 17.38+/-2.67, which was reduced to 6.13+/-1.46 after the intervention (p<0.0001) compared to scores of 18.83+/-2.86 and 6.83+/-2.79, respectively, in the ART group (p<0.0001). The mean urokinase dose was 2.07+/ 0.44 million IU in the LIT group versus 0.70+/-0.36 million IU in the ART group (p<0.0001). The mean duration of therapy was 11.45+/-2.94 hours in the LIT group versus 3.37+/-1.41 hours in the ART group (p<0.0001). No significant difference in relative Miller score improvement was observed. CONCLUSION: By accelerating the fragmentation of thrombus, ART plus adjunctive low-dose urokinase seems to be more rapidly effective compared to LIT. ART achieves both rapid cardiovascular relief and reduces the dose of thrombolytic agent necessary in patients with massive pulmonary embolism. PMID- 15823070 TI - Thrombolytic therapies: the current state of affairs. AB - Thrombotic occlusive diseases are manifested in several disorders that have significant morbidity and mortality, including acute myocardial infarction, pulmonary embolism, deep venous thrombosis, and cerebrovascular accidents. This review summarizes the recently published literature covering thrombolytic therapies in these diseases, with particular attention to comparisons between the fibrin-specific tissue plasminogen activators (alteplase, reteplase, and tenecteplase) and the nonfibrin-specific activators (streptokinase or urokinase plasminogen activator). These agents act to convert plasminogen to plasmin, which in turn cleaves fibrin as part of the lysis process. Fibrin-specific activators were anticipated to be more efficacious and safer than nonspecific agents in thrombolytic occlusive diseases because of their pathophysiologically restricted mechanism of action. However, the fibrin-specific activators also lyse physiological hemostatic plugs, which can result in costly adverse events. Efficacy of fibrin-specific tissue plasminogen activators has been shown to be generally equivalent, with similar mortality rates compared with nonspecific agents; however, fibrin-specific agents may be associated with an increased incidence of intracerebral hemorrhage and with increased costs. Therefore, it appears that given equivalent efficacy, nonfibrin-specific activators, such as streptokinase or urokinase, may be a safer choice in many thrombotic situations. PMID- 15823069 TI - A new mechanical device for declotting of hemodialysis access grafts: first clinical experience. AB - PURPOSE: To review the early clinical experience with a new mechanical thrombectomy device for declotting polytetrafluoroethylene (PTFE) hemodialysis access (HDA) grafts. METHODS: The XTD is a 6-F introducer-compatible device that pulverizes clot with a rotating curved tip driven by a flexible spiral shaft. Fragments are aspirated into a collection container. Fifty patients (32 women; mean age 66 years) with clotted 6-mm polytetrafluoroethylene HDA grafts were treated at 2 medical centers in a study spanning 22 months. In all, 59 procedures were performed on an outpatient basis under monitored conscious sedation. Ancillary techniques were used as needed to treat the underlying cause of the clot. Technical success was defined on a per-procedure basis as removal of sufficient thrombus to visualize the underlying disease without major device related complications. RESULTS: Technical success was 100%; no device-related adverse events occurred. Immediate clinical success was achieved in 47/59 (80%) cases. Continued clinical success in survivors was 52% (30/58) at 1 month and 46% (26/56) at 3 months. Eliminating from analysis a high-risk patient subgroup (reocclusions, chronically clotted or unsalvageable grafts, untreatable central venous stenosis, and venous anastomoses resistant to wire/catheter passage) increased the 3-month clinical success to 59% (22/37). CONCLUSIONS: The XTD is a promising device capable of safely and effectively declotting PTFE grafts. Further investigation is warranted. PMID- 15823071 TI - Impact of the rapid-exchange versus over-the-wire technique on procedural complications of renal artery angioplasty. AB - PURPOSE: To investigate whether the use of rapid-exchange (RX) systems adds to the safety of percutaneous renal artery procedures compared to the conventional over-the-wire (OTW) technique. METHODS: The interventional registry in our department was interrogated to identify patients who underwent plain balloon angioplasty and/or stent implantation for >60% renal artery stenosis and intractable hypertension or decreasing renal function between 1998 and 2004. In this time period, 63 consecutive patients (36 men; mean age 67 years, range 57 80) underwent 78 renal artery angioplasty procedures. The first 46 procedures were done using a transfemoral OTW technique via 7-F sheaths; the following 32 procedures were performed with an RX system via a transfemoral 6-F access. Duration of fluoroscopy, amount of contrast agent, course of serum creatinine, and complications were compared between OTW and RX approaches. RESULTS: Duration of fluoroscopy (median 13.1 versus 11.1 minutes, p=0.099) and primary technical success (94% versus 97%, p=0.64) were not significantly different between the OTW and RX approaches, but significantly less contrast agent was needed with the RX system (median 215 versus 140 mL, p<0.001). Complications, all minor, occurred significantly more often with the OTW (11/46, 24%) compared to the RX system (2/32, 6%; p=0.040) and included misplaced stents, prolonged severe hyper/hypotension, transient renal impairment, and puncture site complications. In particular, the rates of increased serum creatinine within 24 hours were higher in the OTW patients compared to the RX group (20% versus 3%, respectively, for >25% increase [p=0.041] and 9% versus 0% for >50% increase [p=0.087]). CONCLUSIONS: Rapid exchange systems seem to add to the safety of percutaneous renal artery interventions. This likely may be due to a variety of causes, including lower doses of contrast medium, shorter duration of fluoroscopy, and smaller device diameters. PMID- 15823072 TI - PTFE-covered self-expanding nitinol stents for the treatment of severe iliac and femoral artery stenoses and occlusions: final results from a prospective study. AB - PURPOSE: To evaluate the technical performance, safety, and 1-year clinical efficacy of polytetrafluoroethylene (PTFE)-covered nitinol stents in the treatment of atherosclerotic iliac and superficial femoral artery (SFA) disease. METHODS: The multicenter, prospective, nonrandomized COVENT study involved 98 patients (70 men; mean age 64+/-10 years) who received PTFE-covered nitinol stents in 107 arteries (60 iliac and 47 SFAs) after predilation. The average lesion length was 50 mm in the SFA and 45 mm in the iliac arteries. Postdilation was performed when necessary. Duplex ultrasound and ankle-brachial index (ABI) were performed at discharge and at 1, 6, and 12 months in follow-up. RESULTS: In total, 130 stents were placed successfully in 97 (99%) of 98 patients. One stent was misplaced during deployment and required subsequent surgical removal. The average stenosis grade was reduced from 98% to 6% in the SFAs and from 96% to 4% in the iliac arteries after covered stent placement. There was a significant rise of the mean ABI from 0.64 at baseline to 0.97 and 0.95 at 1 and 12 months, respectively (p<0.001). There were 7 primary covered stent occlusions (6.5% of 107 stented lesions: 3 not treated, 2 bypassed, 2 dilated or stented) and 5 (4.7%) recurrent in-stent occlusions (1 bypassed, 2 dilated, 2 untreated) during the 1-year follow-up. Primary patency rates were 92% at 6 months and 89.8% at 12 months for the entire cohort. Secondary patency rates were 98% and 95.6%, respectively. No statistically significant differences were observed in the primary patency rates for the SFAs (89.3% at both 6 and 12 months) versus the iliac arteries (94.3% at 6 months and 90.7% at 12 months). CONCLUSIONS: Primary implantation of PTFE-covered nitinol stents in the iliac and superficial femoral arteries is technically feasible, safe, and effective, with excellent 1-year patency. PMID- 15823073 TI - Crimping and repositioning of a maldeployed balloon-expandable arterial stent using a gooseneck snare. AB - PURPOSE: To describe a technique for repositioning a fully deployed iliac stent from the infrarenal aorta into the common iliac artery (CIA). CASE REPORT: A 58 year-old man was undergoing treatment for a significant right CIA stenosis when a 7x24-mm Palmaz Genesis medium stent was mistakenly deployed in the infrarenal aorta. With the stent still over the guidewire, an 8x60-mm balloon catheter was placed coaxially in the stent. Via a left groin access, a 6-F vascular sheath was introduced retrograde, and a 2.5-cm Amplatz gooseneck snare was advanced into the infrarenal abdominal aorta and pulled back over the stent. The snare was tightly closed to crimp the stent onto the collapsed balloon; this maneuver was repeated several times until the stent was contracted along its entire length. The balloon/stent assembly was carefully pulled back into the right CIA, and the stent was deployed across the target lesion, although there was overlap of the left CIA. Color duplex sonography at 1 year showed no signs of significant iliac arterial stenoses on either side. The patient reported no claudication. CONCLUSIONS: Using a gooseneck snare, fully deployed balloon-expandable iliac stents can be recrimped on a balloon. PMID- 15823074 TI - Rapid progression of discrete type A intramural hematoma: prevention of a "procedure-related" complication by intraoperative transesophageal echocardiography. AB - PURPOSE: To report a case illustrating the utility of transesophageal echocardiography (TEE) before planned stent-graft placement for chronic type B aortic dissection. CASE REPORT: A 64-year-old man with acute aortic syndrome and an 8-year-old interposition graft in the distal aortic arch for acute type B dissection was referred for dissection of the descending thoracic aorta down to the aortic bifurcation; the false lumen was dilated to 65 mm and was partially thrombosed. The ascending aorta showed discrete, eccentric, 4-mm wall thickening that was not considered clinically significant. Stent-graft closure of the entry tear in the proximal descending thoracic aorta was elected. However, as the endovascular procedure was about to commence, TEE showed striking eccentric thickening of the aortic wall of up to 18 mm. The endovascular procedure was stopped, as it was decided to urgently replace the ascending aorta. The next day, the patient underwent successful ascending aortic replacement and simultaneous antegrade stent-graft implantation over the descending thoracic aortic entry tear via the open aortic arch. The postoperative course was uncomplicated, and the patient was discharged 19 days after surgery. He remains well at 6 months after the procedure. CONCLUSIONS: Our case demonstrates that dissection of the ascending aorta may occur not only due to endograft-induced intimal injury, but may also occur due to underlying but undiagnosed or underestimated disease of the ascending aorta or arch. Besides procedural guidance, intraoperative TEE is a useful tool to detect such disease to avoid subsequent "procedure-related" complications. PMID- 15823075 TI - Stenting of a left main coronary artery dissection and stent-graft implantation for acute type a aortic dissection. AB - PURPOSE: To report successful endovascular repair of Stanford type A acute aortic dissection associated with a proximally extended dissection of the left main coronary artery. CASE REPORT: A 71-year-old man presented with acute type A aortic dissection. One day after admission, dissection of the left main coronary artery accompanied by severe myocardial ischemia prompted Palmaz stent placement. Three days later, a customized stent-graft was placed across the entry site of the dissection in the descending aorta. The false lumen in the ascending aorta, transverse arch, and the descending thoracic aorta thrombosed, and the left coronary artery remained patent. At 14 months after the procedures, the patient is doing well and has had no cardiac event. CONCLUSIONS: This staged procedure may be one option for the management of acute type A aortic dissection complicated by coronary artery dissection. PMID- 15823076 TI - A word of caution on the use of high-dose MR contrast media in patients with renal failure. PMID- 15823077 TI - Technique for converting the Excluder stent-graft to an aortomonoiliac system. PMID- 15823078 TI - Kissing-balloon angioplasty with a new low-profile catheter for the popliteal trifurcation. PMID- 15823079 TI - Can suprarenal fixation be the cause of a new renal artery aneurysm remote from an aortic endograft? PMID- 15823080 TI - Is flow reversal the best method of protection during carotid stenting? PMID- 15823082 TI - How does a cell repair damaged DNA? AB - DNA in living cells is constantly subjected to different chemical and physical factors of the environment and to cell metabolites. Some changes altering DNA structure occur spontaneously. This raises the potential danger of harmful mutations that could be transmitted to offspring. To avoid the danger of mutations and changing genetic information, a cell is capable to switch on multiple mechanisms of DNA repair that remove damage and restore native structure. In many cases, removal of the same damage may involve several alternative pathways; this is very important for DNA repair under the most unfavorable conditions. This review summarizes data about all known mechanisms of eukaryotic DNA repair including excision repair (base excision repair and nucleotide excision repair), mismatch repair, repair of double-strand breaks, and cross-link repair. Special attention is given to the regulation of excision repair by different proteins--proliferating cell nuclear antigen (PCNA), p53, and proteasome. The review also highlights problem of bypassing irremovable lesions in DNA. PMID- 15823083 TI - Molecular cloning of Kunitz-type proteinase inhibitor group B genes from potato. AB - Eighteen clones representing copies of four Kunitz-type proteinase inhibitor group B genes (PKPI-B) obtained by polymerase chain reaction cloning of potato (Solanum tuberosum L. cv. Istrinskii) genomic DNA were sequenced and analyzed. Three new genes were found and named PKPI-B1, PKPI-B2, and PKPI-B10: these were represented by five, two, and seven clones, respectively. The remaining four clones corresponded to the formerly characterized PKPI-B9 gene. These data show that at least four PKPI-B encoding genes are harbored in the genome of potato cv. Istrinskii. Their analysis suggests that variability of PKPI-B encoding genes in potato is limited and could be explained by cross-hybridization events in the ancestor forms rather than by random mutagenesis. PMID- 15823084 TI - Digestive proteinases of yellow mealworm (Tenebrio molitor) larvae: purification and characterization of a trypsin-like proteinase. AB - A new trypsin-like proteinase was purified to homogeneity from the posterior midgut of Tenebrio molitor larvae by ion-exchange chromatography on DEAE-Sephadex A-50 and gel filtration on Superdex-75. The isolated enzyme had molecular mass of 25.5 kD and pI 7.4. The enzyme was also characterized by temperature optimum at 55 degrees C, pH optimum at 8.5, and K(m) value of 0.04 mM (for hydrolysis of Bz Arg-pNA). According to inhibitor analysis the enzyme is a trypsin-like serine proteinase stable within the pH range of 5.0-9.5. The enzyme hydrolyzes peptide bonds formed by Arg or Lys residues in the P1 position with a preference for relatively long peptide substrates. The N-terminal amino acid sequence, IVGGSSISISSVPXQIXLQY, shares 50-72% identity with other insect trypsin-like proteinases, and 44-50% identity to mammalian trypsins. The isolated enzyme is sensitive to inhibition by plant proteinase inhibitors and it can serve as a suitable target for control of digestion in this stored product pest. PMID- 15823085 TI - Cloning and expression of mistletoe lectin III B-subunit. AB - Aqueous extracts of mistletoe (Viscum album L.) contain toxic proteins (lectins) MLI (viscumin), MLII, and MLIII. We previously cloned the gene encoding MLIII precursor. In the present study, a gene fragment encoding the carbohydrate binding subunit of mistletoe toxic lectin MLIII was cloned and expressed in Escherichia coli cells. The structure and immunochemical properties of recombinant MLIII B-subunit were investigated using a panel of monoclonal antibodies against ML-toxins. Sugar-binding activity of recombinant MLIII B subunit was determined by ELISA. Amino acid sequence analysis of the cloned MLIII compared with known mistletoe toxins and other ribosome-inactivating type II proteins (ricin, abrin a, and nigrin b B-subunits) revealed essential features of the recombinant MLIIIB primary structure that could determine sugar specificity of the lectin as well as immunomodulating and anti-tumor properties of mistletoe extracts. PMID- 15823086 TI - Effect of salicylic acid on protein composition of Tatar buckwheat Fagopyrum tataricum calluses with different ability for morphogenesis. AB - The effect of salicylic acid on the content of soluble proteins and individual polypeptides in Tatar buckwheat Fagopyrum tataricum calluses differing in ability for morphogenesis was studied. Changes in the protein composition of the calluses cultivated in the dark and in the light indicated the higher sensitivity of the non-morphogenic callus. Different response of callus cultures to salicylic acid and conditions of cultivation (light, darkness) is suggested to be associated with the antioxidant defense system, which is, in particular, characterized by the hydrogen peroxide content in the calluses. Salicylic acid increased the H2O2 content in non-morphogenic calluses more strongly than in morphogenic calluses, and the difference was more significant for the calluses cultivated in the light. PMID- 15823087 TI - Inhibition of peroxidase-catalyzed oxidation of 3,3 ,5,5 -tetramethylbenzidine by aminophenols. AB - Peroxidase-catalyzed oxidation of 3,3 ,5,5 -tetramethylbenzidine (TMB) was inhibited by o-aminophenol (AP), 2-amino-4-tert-butylphenol (ATBP), 2-amino-4,6 di-tert-butylphenol (ADTBP), and 4-tert-butylpyrocatechol (TBP). Inhibitors were characterized by inhibition constant K(i) and stoichiometric coefficient f, the number of radicals terminated by one inhibitor molecule. The most efficient inhibitor is ADTBP characterized by K(i) = 36 microM in 0.015 M phosphate citrate buffer, pH 6.0, at 20 degrees C. According to their antiradical efficiency, the studied inhibitors can be arranged as follows: ADTBP > ATBP > AP > TBP. The role of the NH(2) group in the inhibitory capacity of aminophenols is discussed. Using gas-liquid chromatography, kinetics of consumption of the initial components and accumulation of the reaction products on peroxidase-catalyzed oxidation of the TMB-TBP pair was studied; the data clarify the stages of a complex process of co oxidation of amines and phenols. PMID- 15823088 TI - Sialoside-binding macrophage lectins in phagocytosis of apoptotic bodies. AB - Elimination of apoptotic bodies is one of the important functions of macrophages. The aim of this work was to study the role of macrophage lectins in this process. Macrophage lectins were probed with neoglycoconjugates Glyc-PAA-fluo where carbohydrate is linked to fluorescein-labeled polyacrylamide (MW 30 kD). It was shown that neoglycoconjugates containing a Neu5Acalpha2-3Gal fragment bound to macrophages isolated from blood of healthy donors. Besides, carbohydrate chains containing the same fragment were revealed on apoptotic bodies. Phagocytosis of apoptotic bodies by macrophages was inhibited with sialooligosaccharide ligands of siglec-5 and MAbs to siglec-5. Thus, siglec-5 expressed on macrophages could participate in phagocytosis of apoptotic bodies. In addition, the role of siglecs in engulfment of apoptotic bodies by tumor-associated macrophages was studied. The phagocytic potency of macrophages isolated from blood of breast cancer patients was lower than engulfment ability of macrophages obtained from healthy donors and depended on tumor degree. Staining of macrophages obtained from blood of tumor patients with sialylated Glyc-PAA-fluo probes was more intense than that of macrophages from healthy donors; phagocytosis of apoptotic bodies by tumor associated macrophages was inhibited by carbohydrates that are known to be ligands for siglecs. PMID- 15823089 TI - Role of thiamine thiol form in nitric oxide metabolism. AB - In alkaline media the thiamine cyclic form is converted into a thiol form (pK(a) 9.2) with an opened thiazole ring. The thiamine thiol form releases nitric oxide from S-nitrosoglutathione (GSNO). Thiamine disulfide, mixed thiamine disulfide with glutathione, and nitric oxide are produced in the reaction. Free glutathione was recorded in small amounts. The concentration of formed nitric oxide agreed well with the concentration of degraded GSNO. The concentration of released nitric oxide was determined under anaerobic conditions spectrophotometrically by production of nitrosohemoglobin. In air, the release of nitric oxide was recorded by the production of nitrite or the oxidation of oxyhemoglobin to methemoglobin. The concentration of the thiol form in the body under physiological pH values (7.2-7.4) did not exceed 1.5-2.0%. We believe that due to the exchange reactions between the thiamine thiol form and S-nitrosocysteine protein residues, nitric oxide can be released and mixed thiamine-protein disulfides are formed. The mixed thiamine disulfides (including thiamine ester disulfides) as well as the thiamine disulfide form are quite easily reduced by low molecular weight thiols to form the thiamine cyclic form with a closed thiazole ring. A possible role of the thiamine thiol form in releasing deposited nitric oxide from low-molecular-weight S-nitrosothiols and protein S-nitrosothiols and in regulation of blood flow in the vascular bed is discussed. PMID- 15823090 TI - Comparative study of carrageenans from reproductive and sterile forms of Tichocarpus crinitus (Gmel.) Rupr (Rhodophyta, Tichocarpaceae). AB - A comparative study of the structure and properties of the sulfated polysaccharides (carrageenans) isolated from the vegetative and reproductive forms of the red alga Tichocarpus crinitus was performed. The polysaccharides were separated into the gelling (KCl-insoluble) and non-gelling (KCl-soluble) fractions by precipitation with 4% KCl. The total content of polysaccharides extracted from the reproductive form of the alga was 1.8-fold more than that extracted from the vegetative form, and in the first case, the gelling polysaccharides mostly accumulated. The gelling polysaccharides from the vegetative form have the highest molecular weight (354 kD). According to the results of FT-IR and 13C-NMR spectroscopy, the gelling polysaccharide fractions from both forms are kappa/beta carrageenans. The differences concern the content of the kappa- and beta-disaccharide units and the presence of a small content of the sulfated disaccharide segments (precursors of the kappa-carrageenans) in the polysaccharide from the reproductive form of the alga. The non-gelling polysaccharide fractions from both forms of the plant are mixtures of sulfated galactans with a low content of 3,6-anhydrogalactose. PMID- 15823091 TI - Engineering of proteolytically stable NADPH-cytochrome P450 reductase. AB - NADPH-cytochrome P450 reductase (CPR) is a membrane-bound flavoprotein that interacts with the membrane via its N-terminal hydrophobic sequence (residues 1 56). CPR is the main electron transfer component of hydroxylation reactions catalyzed by microsomal cytochrome P450s. The membrane-bound hydrophobic domain of NADPH-cytochrome P450 reductase is easily removed during limited proteolysis and is the subject of spontaneous digestion of membrane-binding fragment at the site Lys56-Ile57 by intracellular trypsin-like proteases that makes the flavoprotein very unstable during purification or expression in E. coli. The removal of the N-terminal hydrophobic sequence of NADPH-cytochrome P450 reductase results in loss of the ability of the flavoprotein to interact and transfer electrons to cytochrome P450. In the present work, by replacement of the lysine residue (Lys56) with Gln using site directed mutagenesis, we prepared the full length flavoprotein mutant Lys56Gln stable to spontaneous proteolysis but possessing spectral and catalytic properties of the wild type flavoprotein. Limited proteolysis with trypsin and protease from Staphylococcus aureus of highly purified and membrane-bound Lys56Gln mutant of the flavoprotein as well as wild type NADPH-cytochrome P450 reductase allowed localization of some amino acids of the linker fragment of NADPH-cytochrome P450 reductase relative to the membrane. During prolong incubation or with increased trypsin ratio, the mutant form showed an alternative limited proteolysis pattern, indicating the partial accessibility of another site. Nevertheless, the membrane-bound mutant form is stable to trypsinolysis. Truncated forms of the flavoprotein (residues 46-676 of the mutant or 57-676 of wild type NADPH-cytochrome P450 reductase) are unable to transfer electrons to cytochrome P450c17 or P4503A4, confirming the importance of the N-terminal sequence for catalysis. Based on the results obtained in the present work, we suggest a scheme of structural topology of the N-terminal hydrophobic sequence of NADPH-cytochrome P450 reductase in the membrane. PMID- 15823092 TI - Study of the hydrogen bond in different orientations of adenine-thymine base pairs: an ab initio study. AB - In order to gain deeper insight into structure, charge distribution, and energies of A-T base pairs, we have performed quantum chemical ab initio and density functional calculations at the HF (Hartree-Fock) and B3LYP levels with 3-21G*, 6 31G*, 6-31G**, and 6-31++G** basis sets. The calculated donor-acceptor atom distances in the Watson-Crick A-T base pair are in good agreement with the experimental mean values obtained from an analysis of 21 high resolution DNA structures. In addition, for further correction of interaction energies between adenine and thymine, the basis set superposition error (BSSE) associated with the hydrogen bond energy has been computed via the counterpoise method using the individual bases as fragments. In the Watson-Crick A-T base pair there is a good agreement between theory and experimental results. The distances for (N2...H23 N19), (N8-H13...O24), and (C1...O18) are 2.84, 2.94, and 3.63 A, respectively, at B3LYP/6-31G** level, which is in good agreement with experimental results (2.82, 2.98, and 3.52 A). Interaction energy of the Watson-Crick A-T base pair is -13.90 and -10.24 kcal/mol at B3LYP/6-31G** and HF/6-31G** levels, respectively. The interaction energy of model (9) A-T base pair is larger than others, -18.28 and 17.26 kcal/mol, and for model (2) is the smallest value, -13.53 and -13.03 kcal/mol, at B3LYP/6-31G** and B3LYP/6-31++G** levels, respectively. The computed B3LYP/6-31G** bond enthalpies for Watson-Crick A-T pairs of -14.4 kcal/mol agree well with the experimental results of -12.1 kcal/mol deviating by as little as 2.3 kcal/mol. The BSSE of some cases is large (9.85 kcal/mol) and some is quite small (0.6 kcal/mol). PMID- 15823093 TI - Thromboxane A2-induced contraction of rat caudal arterial smooth muscle involves activation of Ca2+ entry and Ca2+ sensitization: Rho-associated kinase-mediated phosphorylation of MYPT1 at Thr-855, but not Thr-697. AB - The signal transduction pathway whereby the TxA2 (thromboxane A2) mimetic U-46619 activates vascular smooth muscle contraction was investigated in de endothelialized rat caudal artery. U-46619-evoked contraction was inhibited by the TP receptor (TxA2 receptor) antagonist SQ-29548, the ROK (Rho-associated kinase) inhibitors Y-27632 and H-1152, the MLCK (myosin light-chain kinase) inhibitors ML-7, ML-9 and wortmannin, the voltagegated Ca2+-channel blocker nicardipine, and removal of extracellular Ca2+; the protein kinase C inhibitor GF109203x had no effect. U-46619 elicited Ca2+ sensitization in a-toxin permeabilized tissue. U-46619 induced activation of the small GTPase RhoA, consistent with the involvement of ROK. Two downstream targets of ROK were investigated: CPI-17 [protein kinase C-potentiated inhibitory protein for PP1 (protein phosphatase type 1) of 17 kDa], a myosin light-chain phosphatase inhibitor, was not phosphorylated at the functional site (Thr-38); phosphorylation of MYPT1 (myosin-targeting subunit of myosin light-chain phosphatase) was significantly increased at Thr-855, but not Thr-697. U-46619 evoked contraction correlated with phosphorylation of the 20 kDa light chains of myosin. We conclude that: (i) U-46619 induces contraction via activation of the Ca2+/calmodulin/MLCK pathway and of the RhoA/ROK pathway; (ii) Thr-855 of MYPT1 is phosphorylated by ROK at rest and in response to U-46619 stimulation; (iii) Thr-697 of MYPT1 is phosphorylated by a kinase other than ROK under resting conditions, and is not increased in response to U-46619 treatment; and (iv) neither ROK nor protein kinase C phosphorylates CPI-17 in this vascular smooth muscle in response to U-46619. PMID- 15823094 TI - Folding pathway of the pyridoxal 5'-phosphate C-S lyase MalY from Escherichia coli. AB - MalY from Escherichia coli is a bifunctional dimeric PLP (pyridoxal 5'-phosphate) enzyme acting as a beta-cystathionase and as a repressor of the maltose system. The spectroscopic and molecular properties of the holoenzyme, in the untreated and NaBH4-treated forms, and of the apoenzyme have been elucidated. A systematic study of the urea-induced unfolding of MalY has been monitored by gel filtration, cross-linking, ANS (8-anilino-1-naphthalenesulphonic acid) binding and by visible, near- and far-UV CD, fluorescence and NMR spectroscopies under equilibrium conditions. Unfolding proceeds in at least three stages. The first transition, occurring between 0 and 1 M urea, gives rise to a partially active dimeric species that binds PLP. The second equilibrium transition involving dimer dissociation, release of PLP and loss of lyase activity leads to the formation of a monomeric equilibrium intermediate. It is a partially unfolded molecule that retains most of the native-state secondary structure, binds significant amounts of ANS (a probe for exposed hydrophobic surfaces) and tends to self-associate. The self-associated aggregates predominate at urea concentrations of 2-4 M for holoMalY. The third step represents the complete unfolding of the enzyme. These results when compared with the urea-induced unfolding profiles of apoMalY and NaBH4-reduced holoenzyme suggest that the coenzyme group attached to the active site lysine residue increases the stability of the dimeric enzyme. Both holo- and apo-MalY could be successfully refolded into the active enzyme with an 85% yield. Further refolding studies suggest that large misfolded soluble aggregates that cannot be refolded could be responsible for the incomplete re-activation. PMID- 15823095 TI - Mammalian Lass6 and its related family members regulate synthesis of specific ceramides. AB - The Lass (longevity-assurance homologue) family members, which are highly conserved among eukaryotes, function in ceramide synthesis. In the mouse, there are at least five Lass family members, Lass1, Lass2, Lass4, Lass5 and the hitherto uncharacterized Lass6. To investigate specific roles for each Lass member in ceramide synthesis, we cloned these five mouse proteins. Overproduction of any Lass protein in cultured cells resulted in an increase in cellular ceramide, but the ceramide species produced varied. Overproduction of Lass1 increased C18:0-ceramide levels preferentially, and overproduction of Lass2 and Lass4 increased levels of longer ceramides such as C22:0- and C24:0-ceramides. Lass5 and Lass6 produced shorter ceramide species (C14:0- and C16:0-ceramides); however, their substrate preferences towards saturated/unsaturated fatty acyl-CoA differed. In addition to differences in substrate preferences, we also demonstrated by Northern blotting that Lass family members are differentially expressed among tissues. Additionally, we found that Lass proteins differ with regard to glycosylation. Of the five members, only Lass2, Lass5 and Lass6 were N glycosylated, each at their N-terminal Asn residue. The occurrence of N glycosylation of some Lass proteins provides topological insight, indicating that the N-termini of Lass family members probably face the luminal side of the endoplasmic reticulum membrane. Furthermore, based on a proteinase K digestion assay, we demonstrated that the C-terminus of Lass6 faces the cytosolic side of the membrane. From these data we propose topology for the conserved Lag1 motif in Lass family members, namely that the N-terminal region faces the luminal side and the C-terminal region the cytosolic side of the endoplasmic reticulum membrane. PMID- 15823096 TI - Expression of functional recombinant von Willebrand factor-A domain from human complement C2: a potential binding site for C4 and CRIT. AB - CRIT (complement C2 receptor inhibitor trispanning) is a newly described transmembrane molecule that is capable of binding C2 via its first extracellular domain (ed1). CRIT competes with C4b for the binding of C2. Previous experiments have suggested that a major binding site for C2 is located on short, almost identical peptide sequences of CRIT-ed1 and the beta-chain of C4. The C2 domains involved in binding, however, remain unknown. We cloned the vWFA (von Willebrand factor-A) domain of C2, as it is a region likely to be involved in interactions with other proteins, and were able to functionally express the 25 kDa human complement C2 vWFA domain (amino acids 224-437). The recombinant vWFA protein fixed on MagneHis Ni-Particles bound C4 in normal human serum. The C4 alpha, beta and gamma chains were separated by SDS/PAGE and purified separately by electro elution. The purified C4 chains were then used in a sandwich ELISA, which showed the vWFA to bind C4 only via the C4beta chain. In a haemolytic assay, the recombinant vWFA protein inhibited complement activation by the classical pathway in a dose-dependent manner by competing with native C2 for binding to C4b. vWFA bound the ed1 peptide of CRIT as well, and specifically to the 11-amino-acid peptide fragment of ed1 that is known to interact with whole C2. These findings show that the vWFA domain is centrally involved in the C2-CRIT and C2-C4b bindings. The cloned vWFA domain will allow us to dissect out the fine interactions between C2 and CRIT or C4b. PMID- 15823097 TI - Identification of a functional hypoxia-responsive element that regulates the expression of the egl nine homologue 3 (egln3/phd3) gene. AB - Low oxygen levels induce an adaptive response in cells through the activation of HIFs (hypoxia-inducible factors). These transcription factors are mainly regulated by a group of proline hydroxylases that, in the presence of oxygen, target HIF for degradation. The expression of two such enzymes, EGLN1 [EGL nine homologous protein 1, where EGL stands for egg laying defective (Caenorhabditis elegans gene)] and EGLN3, is induced by hypoxia through a negative feedback loop, and we have demonstrated recently that hypoxic induction of EGLN expression is HIF-dependent. In the present study, we have identified an HRE (hypoxia response element) in the region of the EGLN3 gene using a combination of bioinformatics and biological approaches. Initially, we isolated a number of HRE consensus sequences in a region of 40 kb around the human EGLN3 gene and studied their evolutionary conservation. Subsequently, we examined the functionality of the conserved HRE sequences in reporter and chromatin precipitation assays. One of the HREs, located within a conserved region of the first intron of the EGLN3 gene 12 kb downstream of the transcription initiation site, bound HIF in vivo. Furthermore, this sequence was able to drive reporter gene expression under conditions of hypoxia in an HRE-dependent manner. Indeed, we were able to demonstrate that HIF was necessary and sufficient to induce gene expression from this enhancer sequence. PMID- 15823098 TI - Pattern of care and impact of participation in clinical studies on the outcome in ovarian cancer. AB - The purpose of this study was to evaluate the pattern and quality of care for ovarian cancer in Germany and analyze prognostic factors with emphasis on characteristics of treating institutions, hospital volume, and participation in clinical trials. This study utilized national survey including patients with histologically proven invasive epithelial ovarian cancer diagnosed in the third quarter of 2001 including descriptive analysis of pattern of surgical care and systemic treatment in early (FIGO I-IIA) and advanced (FIGO IIB-IV) ovarian cancer and both univariate and multivariate analysis of prognostic factors. One third of all patients diagnosed in the third quarter of 2001 in Germany, 476 patients, were included. Standard care according to German guidelines was provided to only 35.5% of patients with early ovarian cancer. Recommended chemotherapy was given to 78% in advanced disease. Multivariate analysis showed advanced stage, poor performance status, comorbidity, ascites, and treatment in an institution not participating in cooperative studies to be associated with inferior survival. Non-participation was associated with an 82% increase of risk (HR = 1.82; 95% CI, 1.27-2.61; P= 0.001). Hospital volume did not affect treatment outcome. Adherence to treatment guidelines showed remarkable variety among German hospitals, indicating options and need for improvement. Selecting an institution that participates in cooperative trials might be an option for individual patients seizing the chance for better quality of care even when individual factors might hamper enrollment in a study. PMID- 15823099 TI - Accuracy of frozen-section analysis in the diagnosis of ovarian tumors: a systematic quantitative review. AB - A quantitative systematic review was performed to estimate the diagnostic accuracy of frozen sections in ovarian tumors. Studies that compared frozen sections and paraffin sections within subjects for diagnosis of ovarian tumors were included. Fourteen primary studies were analyzed, which included 3 659 women. For benign ovarian vs borderline/malignant tumor cases, the occurrence of a positive frozen-section result for benignity (pooled likelihood ratio [LR], 8.7; 95% confidence interval [CI], 7.3-10.4) and posttest probability for benign diagnosis was 95% (95% CI, 94-96%). A positive frozen-section result for malignant vs benign diagnosis (pooled LR, 303; 95% CI, 101-605) increased the probability of ovarian cancer to 98% (95% CI, 97-99%). In borderline vs benign ovarian tumor cases, a positive frozen-section result (pooled LR, 69; 95% CI, 45 106) increased the probability of borderline tumors to 79% (95% CI, 71-85%). In borderline vs malignant ovarian tumor cases, a positive frozen-section result (pooled LR, 18; 95% CI, 13-26) increased the probability of borderline tumors to 51% (95% CI, 42-60%). We conclude that diagnostic accuracy rates for frozen section analysis is high for malignant and benign ovarian tumors, but the accuracy rates in borderline tumors remain relatively low. PMID- 15823100 TI - Understanding and treating depression among cancer patients. AB - Cancer is a devastating disease. Awareness of this disorder tremendously impacts an individual's life, creating a host of changes to occur. Depression is one concern that many cancer patients experience. It is important to make an accurate and complete assessment of depressive symptoms. Furthermore, a heightened awareness of patients' culture and their perceptions is vital to their assessment, diagnosis, and treatment of depression. In cases of clinical depression, appropriate antidepressants prescribed along with the treatment for cancer are significant for the patient. Palliative counseling for patients and supportive counseling for their family are recommended as part of a treatment plan. PMID- 15823101 TI - Retrospective review: re-treatment of patients with ovarian cancer with carboplatin after platinum resistance. AB - The objective of the analysis was to determine the effectiveness of re-treating patients with ovarian cancer, primary peritoneal cancer, and fallopian tube cancer with carboplatin after being deemed platinum resistant. From a database period January 1, 1996, to December 12, 2002, 34 patients were identified who received nonplatinum agents before resuming treatment with carboplatin. The median age was 65 years, and a median of two nonplatinum chemotherapy (range 1-5) prior to re-treatment with carboplatin was received. The median platinum-free interval from the time platinum was last received to re-treatment with carboplatin was 15.2 months (95% confidence interval [CI] 12.6-17.9; range 6.2 47.0). A median number of four cycles of carboplatin (range 1-11) was received. Two patients (5.9%) achieved partial response, while 21 patients (61.7%) achieved stable disease. The median time to progression for these 23 patients after re treatment with carboplatin was 5.7 months (95% CI 5.2-6.3; range 1.8-15.3). Twenty-seven patients have died, and all patients have progressed. Seven patients are still receiving salvage therapy. The median overall survival from the time deemed to be platinum resistant is 23.2 months (95% CI 20.1-26.4). Patients who have been deemed platinum resistant may still benefit from platinum re-treatment after an interval of treatment with nonplatinum agents. PMID- 15823102 TI - Neoadjuvant chemotherapy in advanced ovarian cancer: a case-control study. AB - The aim of this study was to compare the outcome of patients with advanced ovarian carcinoma treated with neoadjuvant chemotherapy (NACT) with those treated conventionally with primary debulking surgery. From 1994 to 2003, all consecutive cases of advanced-stage epithelial ovarian carcinoma treated with NACT at the University of Bari were identified. A well-balanced group of women who underwent primary debulking surgery followed by platinum-based chemotherapy was selected as controls. Kaplan-Meier and Cox proportional hazards analyses were used to determine the predictors for survival. Thirty women with advanced-stage epithelial ovarian carcinoma were treated with NACT and compared to 30 patients who underwent primary debulking surgery. Patients in the NACT were significantly older and had a poorer performance status compared to the controls. However, no statistical difference was observed in overall disease-specific survival (P= 0.66) and disease-free survival (P= 0.25) between the two groups. Although patients in the NACT group are significantly older and have a poorer performance status, this treatment modality does not compromise survival. Prospective randomized trials comparing NACT to conventional treatment to determine the quality of life and cost/benefit outcomes are now appropriate for women presenting advanced epithelial ovarian cancer. PMID- 15823103 TI - Carboplatin hypersensitivity induced by low-dose paclitaxel/carboplatin in multiple platinum-treated patients with recurrent ovarian cancer. AB - We report five cases of carboplatin (CBDCA) hypersensitivity after weekly low dose paclitaxel (60 mg/m2)/CBDCA (area under the concentration curve = 2) therapy in patients with recurrent ovarian cancer receiving multiple platinum-based chemotherapy. Paclitaxel and CBDCA therapy was indicated as second-line chemotherapy in one patient and as third line in four patients with recurrent disease. The range of previously administered total CBDCA was 2582-9589 mg, and the CBDCA area under the concentration curve of the first treatment exhibited appropriate intensity (mean, 1.92 +/- 0.10; range, 1.76-2.10) in all patients. However, one patient exhibited severe hypersensitivity reactions including cardiac arrest and apnea, and another four patients developed eruptions, hypotension, and tachycardia soon after administration of CBDCA. Our report suggested that CBDCA hypersensitivity was correlated with the total dose of previously administered platinum agents and that CBDCA should be excluded in patients who have received multiple platinum-based chemotherapy, even in platinum sensitive cases, because CBDCA hypersensitivity can occur even with low-dose CBDCA administration. PMID- 15823104 TI - A randomized prospective study of comparison of reservoir ports versus conventional vascular access in advanced-stage ovarian carcinoma cases treated with chemotherapy. AB - Vascular access ports were developed to overcome many of the problems associated with limited peripheral access, combined with the need for frequent venipuncture, in oncology patients receiving long-term intensive therapy. In this study, we compared the effectivity and acceptability of vascular access port with conventional needle application together with complication rates in ovarian cancer patients. Advanced-stage ovarian carcinoma cases under chemotherapy treatment were equally randomized into two groups, implantable vascular access ports applied to one group (22 cases) and conventional vascular access applied to the other (38 cases) as a control group. Anteroposterior thoracic X-rays of implantable port-applied cases were taken before and after the application. Vortex reservoir ports (Horizon Medical Products, Inc., Manchester, GA) were used in the application to the subclavian vein. Classic peripheral venipuncture method (Medikit), Mediflon(trade mark) IV cannula with PTFE radiopaque catheter and injection valve, Eastern Medikit Ltd, Gurgaon, Haryana, India) was used in the control group. Vascular accesses of all cases were controlled just after the application, 12 h after the application, and during each drug or intravenous fluid application. Mean port insertion time was 26.3 min. Total port occlusion was observed in two of the port-applied cases (11.7%) and partial port occlusion was observed in five of the port-applied cases (29%). Heparin and saline combination was used in order to open the port tip, in five cases, two with total occlusion and three with partial occlusion. Infection was observed in only one case (5%) to whom appropriate therapy was given, and the port was taken out. Ports of two cases were also taken out because of skin dehiscence. No change in port tip position was observed in any of the cases. Total occlusion was observed in 16 of the 38 cases (42.1%) with conventional vascular access. In 12 cases (31.5%), a need arose to change the conventional vascular access. No vascular access was found in 13 of the 38 cases (34.2%). Application of reservoir ports especially to cases with advanced-stage carcinomas, under chemotherapeutic drug treatment, leads to minimal anxiety for the patient and his/her family and minimal risk of physical trauma to the patient with only one vascular access. Reservoir ports occlude or cause infection to a lesser extent than classic vascular access methods. Occlusion or infection rates of reservoir ports are statistically significant, lower than those of classic venipuncture. PMID- 15823105 TI - Intraperitoneal hyperthermic chemotherapy in ovarian cancer. AB - We investigated the effect of intraperitoneal hyperthermic perfusion chemotherapy as consolidation therapy in stage IIIB-IIIC ovarian cancer, following cytoreductive surgery and systemic chemotherapy (cisplatin-cyclophosphamide--six cycles). Disease-free survival, overall survival, and side effects were compared with a control group of patients who refused a second-look surgery and intraperitoneal chemotherapy. In a multicenter prospective trial, 29 patients with complete or optimal cytoreductive surgery and systemic treatment were included in the consolidation group and received intraperitoneal hyperthermic perfusion chemotherapy. Patients were recruited between January 1991 and December 1997. The intraperitoneal hyperthermic perfusion was performed with open-abdomen technique, using physiologic solution containing cisplatin 100 mg/m2, for 60 min in hyperthermic phase (41-43 degrees C). Intraperitoneal hyperthermic perfusion chemotherapy was locally and systemically well tolerated. The consolidation therapy group showed a better 5-year survival rate and lower recurrent disease rate, but differences were not statistically significant. Our results suggest that intraperitoneal hyperthermic perfusion chemotherapy is a feasible, well tolerated, and promising alternative as consolidation therapy in ovarian cancer. PMID- 15823106 TI - Predictive value of serum interleukin-8 levels in ovarian cancer patients treated with paclitaxel-containing regimens. AB - Previous findings showed that paclitaxel induces interleukin-8 (IL-8) transcription and secretion in ovarian cancer cells in vitro. We hypothesized that paclitaxel treatment, which is a standard care for ovarian cancer patients, may increase the secretion of IL-8, resulting in the elevated serum IL-8 levels. In this study, we investigated the relationship between paclitaxel exposure and IL-8 levels of an ovarian and a breast carcinoma cell line in vitro and serums of patients with ovarian carcinoma. Both MDAH 2774 ovarian and MCF-7 breast carcinoma cell lines were sensitive to paclitaxel-mediated cytotoxicity. However, supernatant levels of IL-8 assessed by enzyme-linked immunosorbent assay before and after treatment with different concentrations of paclitaxel were significantly lower in MCF-7 than in MDAH 2774. Serum IL-8 levels were measured in serum samples from patients with ovarian carcinoma before and after paclitaxel containing treatment regimens. Forty-eight patients were included in the study. The basal level of IL-8 after paclitaxel-containing treatment was found to be significantly higher in the serums of patients who had high tumor burden than in patients who had optimal debulking surgery and low tumor burden. These data strongly suggest that IL-8 may be an important predictive marker for tumor volume as well as sensitivity to paclitaxel. PMID- 15823107 TI - A novel early-stage orthotopic model for ovarian cancer in the Fischer 344 rat. AB - The purpose of our study was to ascertain the progression of metastases in a novel ovarian cancer model designed to mimic early-stage disease by utilizing an orthotopic injection technique. Female Fischer 344 rats were injected with either 10(4) or 10(5) NuTu-19 cells by intraperitoneal or orthotopic injection. Peritoneal washings and histologic specimens were examined to correlate the incidence and extent of tumor growth. In a second phase, orthotopic injections of 10(2) and 10(3) cells were compared to that of 10(4) cells. Progression of ovarian cancer was observed by gross and microscopic examinations in both intraperitoneal and orthotopic models. Pelvic extension and abdominal adhesions uniquely characterized the orthotopically injected animals. Numbers of identifiable metastases declined with lower cell inocula, confirming that early stage disease was extended to at least 14 days with 10(2) NuTu-19 cells. The orthotopic ovarian cancer model emulates early disease with the initiation of a primary tumor that is localized within the inherent microenvironment. The orthotopic model offers a clinically relevant alternative for future cancer research that allows for the investigation of therapeutic strategies against early stages of the disease process. PMID- 15823109 TI - Comparing the genetic changes detected in the primary and secondary tumor sites of ovarian cancer using comparative genomic hybridization. AB - Our objective was to compare the genetic abnormalities in the primary tumors of epithelial ovarian cancer and their associated secondary peritoneal implants using comparative genomic hybridization (CGH). CGH was performed on seven apparent stage III ovarian serous cancer cases. Dissected tissue samples from the primary tumor and from the metastatic peritoneal implant were obtained at initial surgical staging and analyzed in each case. We used CGH as this technique allows the entire genome of the tumor to be examined simultaneously for chromosomal imbalances without the need for tissue culture or targeting of specific loci. The chromosomal abnormalities detected in the distinct sites were then reviewed and compared. CGH studies were successful in all 14 samples from the seven patients. The analysis revealed chromosomal aberrations in six patients with certain repeated changes as amplification of 1q, 2p, 2q, 3q, 6q, 8q, and 12p and underrepresentation of 18q and X chromosomes. Comparing the genomes of the primary tumors with the metastatic samples showed four cases with a balanced metastatic CGH profile while the primary site was aberrant. Greater chromosomal complexity associated with the primary site was detected in two other patients. In one case, both primary and secondary sites had no detectable chromosomal imbalances. The cytogenetic patterns in six of the seven primary tumors showed complex karyotypic changes, unlike the inconsistent findings that were associated with the secondary sites. The chromosomes of the secondary sites expressed either normal genomes or fewer genetic aberrations. Such genomic heterogeneity between the primary and secondary sites may indicate that the secondary peritoneal implants are de novo carcinogenesis occurrences. The results may support the concept that at least part of advanced ovarian cancer is a multicentric disease in the early stages. Further genetic studies are needed to reassess this assumption. PMID- 15823108 TI - Increased cyclooxygenase-2 (COX-2) and P-glycoprotein-170 (MDR1) expression is associated with chemotherapy resistance and poor prognosis. Analysis in ovarian carcinoma patients with low and high survival. AB - The aim of the study is to test the prognostic value of cyclooxygenase-2 (COX-2) and P-glycoprotein in relation to responsiveness to chemotherapy in ovarian carcinoma patients with "shorter and longer" survival. We evaluated 52 ovarian carcinomas homogeneous for stage, histologic type, grade of differentiation, and surgical and chemotherapeutic treatment. Twenty-eight of the patients had died of progression of disease no later than 2 years after primary surgical treatment, while 24 patients were alive with no evident disease 5 years after primary surgical treatment. In logistic regression analysis, COX-2 and P-glycoprotein, when analyzed one by one, are significant (P= 0.017 and P < 0.0005, respectively). P-glycoprotein is correlated with COX-2 (P= 0.008, Fisher's exact test); moreover, both COX-2 and the P-glycoprotein are correlated with clinical response to chemotherapy (P= 0.022 and P < 0.0005, respectively, Chi-square test). Our data suggest that COX-2 and P-glycoprotein may have prognostic significance in advanced ovarian serous carcinoma. The COX-2 and the P glycoprotein overexpressions are correlated to one another and both with a progression of disease during the first-line chemotherapy. The administration of a COX-2 inhibitor in association with chemotherapy in ovarian carcinoma patients may improve the tumor chemosensibility and the overall survival. PMID- 15823110 TI - Information given to women invited for cervical screening: results of two postal surveys seven years apart. AB - In South Staffordshire, England, we compared women's views on information provided to them at different stages of the cervical screening program in 1994 with that provided in 2001. An age-stratified random sample of women aged 20-64 years who had a cervical smear taken between January and March 1994 (3856) or between January and March 2001 (4057) were sent postal questionnaires in June 1994 and July 2001, respectively. Response rates in 1994 (3124/3856, 81%) and 2001 (3288/4057, 81%) were similar. Compared to 1994, the proportion of women who thought the invitation letter was clear to read in 2001 increased (70% vs 98%, P < 0.0001); however, letters were thought to be less reassuring in 2001 compared to 1994 (P < 0.0001). In both study periods, 66% of women reported that the procedure was explained to them before the smear was taken. A greater proportion of women received their results by letter in 2001 compared to 1994 (57% vs 41%, P < 0.0001); however, 49% of women waited >4 weeks to receive their results in 2001 compared to 26% in 1994 (P < 0.0001). Bivariate analysis suggests that responses were age related, with older women (> or =45 years) experiencing poorer information provision. The issues highlighted by this study deserve further investigation in other areas. PMID- 15823111 TI - Sentinel node detection with radiocolloid lymphatic mapping in early invasive cervical cancer. AB - We assessed the feasibility of sentinel lymph node detection using technicium-99 radiocolloid lymphatic mapping for predicting lymph node metastases in early invasive cervical cancer. Thirty patients with cervical cancer (stages IA2-IIA) underwent preoperative lymphoscintigraphy using technicium-99 intracervical injection and intraoperative lymphatic mapping with a handheld gamma probe. After dissection of the sentinel nodes, the standard procedure of pelvic lymph node dissection and radical hysterectomy was performed as usual. The sentinel node detection rate was 100% (30/30). There were seven (23.3%) cases of microscopic lymph node metastases on pathologic analysis. All of them had sentinel node involvement. Therefore, the sensitivity of sentinel node identification for prediction of lymph node metastases was 100%, and no false negative was found. Preoperative lymphoscintigraphy, coupled with intraoperative lymphatic mapping, located the sentinel nodes accurately in our study patients. This sentinel node detection method appears to be feasible for predicting lymph node metastases. PMID- 15823112 TI - Human papillomavirus status in advanced cervical cancer: predictive and prognostic significance for curative radiation treatment. AB - Human papillomavirus (HPV) infection plays a major role in oncogenesis of squamous cell carcinoma of the cervix. This study was performed to investigate if HPV status and E2 gene integrity are prognostic parameters for clinical outcome and predictive for radiation response. Forty women with locally advanced cervical cancer treated with curative radiotherapy were analyzed for HPV infection and E2 gene integrity by multiplex polymerase chain reaction. Statistical analyses were performed for overall survival, disease-free survival (DFS), local progression free survival, and treatment response (clinical complete remission). Twenty-eight (70%) of 40 carcinomas were HPV positive. The only significant factor for a better overall survival, DFS, and local progression-free survival was HPV positivity (P < 0.02, P= 0.02, and P < 0.05, log-rank, respectively). HPV positive tumors had a significantly better clinical complete remission (67% vs 33%, P= 0.04, Fisher's exact test). An intact E2 gene region showed a trend for a better DFS (P= 0.1, log-rank). This study reveals HPV as an independent prognostic parameter for outcome and radiation response. Integration of the virus genome into host cell DNA might be a molecular target to determine the treatment response of HPV-positive cancers. PMID- 15823113 TI - Adjuvant radiotherapy in carcinomas of the uterine cervix: the prognostic value of hemoglobin levels. AB - Anemia has been associated with a poorer treatment response and reduced survival in women undergoing primary radiotherapy (RT) or radiochemotherapy for advanced cervical carcinoma. This study aimed to determine the influence of anemia on outcome in patients with cervical carcinoma undergoing adjuvant RT. Medical records were reviewed for 183 cervical cancer patients who had received adjuvant RT because of risk factors after radical surgery (n= 109) or inadequate primary surgery (simple hysterectomy; n= 74). Kaplan-Meier and Cox regression analyses were used to study hemoglobin levels before and during adjuvant RT in relation to recurrence-free and overall survival. Hemoglobin values > or =11 g/dL were considered normal, while those <11 g/dL indicated anemia. Hemoglobin levels before RT influenced significantly overall survival and recurrence-free survival across the whole group (overall survival--log rank(all patients)= 7.5; df = 1; P= 0.006). However, subgroup analysis showed that the observed difference was mainly due to the group of women who had undergone inadequate primary surgery (overall survival--log rank(inadequate surgery)= 10.8; df = 1; P= 0.001). Multifactorial regression analyses comparing hemoglobin before RT with grading and tumor stage confirmed the prognostic value of hemoglobin values. Maintaining normal hemoglobin values before and during adjuvant RT seems to be important, especially in patients who have had inappropriate simple hysterectomy, which may resemble a therapeutic situation. PMID- 15823114 TI - Evaluation of cytologic screening results of the cervix. AB - The aim of this study was to evaluate retrospectively cytologic screening for cancers and precancerous lesions of the cervix and to research whether biopsy is overused among women with a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS). We examined 28 469 smears obtained over 5 years. The patients were referred when the result of the smear was abnormal for a colposcopic biopsy, endocervical curettage, or a repeat Papanicolaou smear. The results of the screening of 28 469 smears are as follows: 699 (2.45%) ASCUS, 67 (0.23%) low-grade squamous intraepithelial lesions (LGSIL), 43 (0.15%) high-grade squamous intraepithelial lesions (HGSIL), 1 epidermoid carcinoma, 31 (0.10%) atypical glandular cells, and 1 adenocarcinoma. Histologic examination of the cervix was done in 119 patients (17.0%) of ASCUS, 13 patients (10.9%) of LGSIL, and 15 patients (12.6%) of HGSIL and cervical carcinoma. In the 119 women with histological examination, histologic examination was generally performed in patients with ASCUS neoplastic Papanicolaou smear, and histologic diagnosis of low-grade, high-grade, or invasive lesion of the cervix was made in 23.5% of women with ASCUS; in these patients, 46.4% were cytologic LGSIL and 53.5 % were cytologic HGSIL and cervical carcinoma. In the aspect of these findings, we concluded that for patients with a cytologic diagnosis of ASCUS, more aggressive interventions should be performed. PMID- 15823115 TI - Small-cell carcinoma of the uterine cervix: a clinicopathologic study of 11 cases. AB - We report the clinical profiles and immunohistochemical features of small-cell carcinoma of the uterine cervix. Eleven cases that we have encountered at the Department of Gynecology, Kitasato University Hospital, between 1971 and 2003 are presented. Of 1370 invasive carcinomas of the uterine cervix, the incidence of small-cell carcinoma was 0.8%. Patient ages ranged between 32 and 65 years, with a mean age of 46.3 years. The clinical stages at diagnosis were Ib in four patients, IIb in three, IIIb in three, and IVb in one. All patients presented with abnormal vaginal bleeding. Two patients who are alive with no evidence of disease for 12 years and 3 years 6 months, while eight patients died of primary carcinoma between 4 and 25 months after treatment. Histopathologic findings showed solid nests with marked peripheral palisading pattern and rosette formation. Small tumor cells with scant cytoplasm demonstrated a very high nuclear/cytoplasm ratio and indistinct cell borders. The nuclei were round to oval and demonstrated increased but fine granular chromatin. Nucleoli were indistinct in all cases. Immunohistochemical findings were positive in 81.8% each for neuron-specific enolase and protein gene product 9.5, 72.7% for synaptophysin, 63.6% for chromogranin A, and 54.5% for neural cell adhesion molecule. All specimens were positive for at least one of the above. In conclusion, small-cell carcinoma of the uterine cervix revealed poor prognosis. Making an accurate diagnosis of small-cell carcinoma before performing treatment is of great significance but often difficult. Immunohistochemical analysis using several kinds of neuroendocrine markers is helpful in establishing the correct diagnosis in addition to focusing on characteristic histo- and cytopathologic features. PMID- 15823116 TI - Antiproliferative activity and toxicity of 2-methoxyestradiol in cervical cancer xenograft mice. AB - 2-methoxyestradiol (2-ME) is considered to be an effective anticancer compound for many types of tumors. We have previously demonstrated that 2-ME inhibits the growth of human cervical cancer HeLaS3 cells in vitro. In this study, we investigated the antitumoral effects of 2-ME on human cervical carcinoma in severe combined immune deficient (SCID) mice. The potential side effects of 2-ME on the SCID mice were also investigated. SCID mice were injected with HeLaS3 cells (3 x 10(6) to 4 x 10(6)/mouse) and a 15-day administration of 2-ME followed after a 1-week cell implantation. Tumor weight, volume, body weight, and blood chemistry were determined. Tumor tissues were examined with an antibody against the proliferative cell nuclear antigen and terminal deoxynucleotidyl transferase mediated dUTP nick end-labeling (TUNEL) staining. Liver, spleen, kidney, heart, and lung were screened by pathologic examinations. 2-ME (75 mg/kg p.o.) inhibited growth of human cervical carcinoma by 34% (P < 0.05) as compared with control. Necrosis was found in both 2-ME-treated and untreated tumor tissues, but the necrotic area was larger in 2-ME-treated mice. A low expression of proliferative cell nuclear antigen and an increased number of apoptotic cells were found in 2 ME-treated tumor sections as compared to those in controls. No significant difference was detected in blood chemistry. In addition, the liver showed hyperplastic Kupffer cells, hydropic swelling of hepatocytes, and liquefactive necrosis. The spleen showed an increased number of megakaryocytes and apoptotic cells after 2-ME treatment. Thus, 2-ME has an antitumor effect on human cervical carcinoma, and it is toxic to liver and spleen in this mouse model. PMID- 15823117 TI - Microsatellite instability is not related to response to cisplatin-based chemotherapy in cervical cancer. AB - This study investigates whether microsatellite instability (MSI) due to defects of the mismatch repair (MMR) system could be associated with response to cisplatin-based neoadjuvant chemotherapy (NACT) and if cisplatin exposure could select MSI-positive cell clones in cervical cancer. Microsatellite analysis was performed by polymerase chain reactions using six microsatellite markers, while hMLH1 protein expression was investigated by immunohistochemistry. We found that 1 tumor out of 20 (5%) NACT-responding patients and 1 tumor out of 18 (6%) nonresponding patients showed MSI. The analysis of tumor specimens collected after NACT revealed no change in the banding pattern as compared to each corresponding pre-NACT tumor at each locus tested. hMLH1 staining was observed in at least > or =80% of cells in all tumors examined except the two exhibiting MSI. Our data showed that MSI due to defects of the MMR system seems not to play a crucial role in the biology of human cervical cancer cells and that MSI seems not to be related to response to chemotherapy. Moreover, cisplatin exposure did not seem to select for MMR-deficient tumor clones in cervical cancer. PMID- 15823118 TI - Epstein-Barr virus plays little role in cervical carcinogenesis in Korean women. AB - We examined whether Epstein-Barr virus (EBV) infection plays a role in cervical carcinogenesis in Korean women. EBV infection was examined using polymerase chain reaction (PCR) with two different primer pairs flanking the BamHI "W" fragment of EBV and by EBV-encoded small RNAs (EBER) in situ hybridization in various histologic types of cervical cancer, including 17 cases of squamous cell carcinoma, 36 cases of adenocarcinoma, and 3 cases of small-cell carcinoma. We also evaluated 20 cases of cervical intraepithelial neoplasia and 20 cases of normal uterine cervix. One case of squamous cell carcinoma and three cases of cervical intraepithelial neoplasia were positive for EBV DNA using PCR, but EBER in situ hybridization analysis showed that none of the PCR-positive cases expressed EBER. EBV DNA was not found using PCR in any of the 20 normal uterine cervices. From our results, EBV infection does not seem to play a role in cervical carcinogenesis in Korean women. PMID- 15823119 TI - Total laparoscopic hysterectomy versus total abdominal hysterectomy for obese women with endometrial cancer. AB - Obesity is common in endometrial cancer and surgery for these patients is challenging. We compared total laparoscopic hysterectomy (TLH) with total abdominal hysterectomy (TAH) with respect to feasibility (operating time, estimated blood loss, length of hospital stay, and conversion to laparotomy) and safety (perioperative morbidity and mortality) in a retrospective analysis of 78 morbidly obese patients with endometrial cancer. Analysis is based on the intention to treat. The intention to treat was TLH in 47 patients and it could be successfully completed in 42 patients (89.4%). The mean weight for all patients was 118.7 kg, with patients in the TLH group weighing more and having higher ASA scores. Mean operating time and estimated blood loss were similar in both groups. Mean postoperative hospital stay was 4.4 (+/-3.9) days in the TLH group and 7.9 (+/-3.0) days in the TAH group (P < 0.0001). Wound infections occurred in 15 of 31 patients (48.4%) in the TAH group and in 1 of 47 patients (2.1%) in the TLH group. All other morbidity, as well as patterns of recurrence and survival were similar in both groups. These data justify a prospective randomized trial comparing TLH with TAH for the treatment of endometrial cancer. PMID- 15823120 TI - Transvaginal ultrasound and diagnostic hysteroscopy as a predictor of endometrial polyps: risk factors for premalignancy and malignancy. AB - The aim of this study is to assess accuracy of transvaginal ultrasound (TVUS) and diagnostic hysteroscopy in diagnosing endometrial polyps and to determine premalignancy and malignancy rates in asymptomatic women. The study was designed to retrospectively analyze 438 women who underwent operative hysteroscopy in a day-care unit when endometrial polyp was suspected after TVUS and diagnostic hysteroscopy. Multivariate logistic regression modeling showed effects of age, previous breast cancer with tamoxifen treatment, and menopause with or without bleeding on pathologic results. The results indicate that positive predictive value of TVUS with diagnostic hysteroscopy was 79.9%. Premalignancy or malignancy occurred in 3.2% and was significantly related to menopause with abnormal bleeding (P < 0.001), which carried a 20-fold higher risk of pathology than any other group. Age was also a risk factor. It was concluded that TVUS with diagnostic hysteroscopy reliably evaluates endometrial polyps. The low incidence of endometrial tumors in asymptomatic (especially premenopausal) women suggests that their operative evaluation may not be cost effective. Larger studies are needed to support this tentative conclusion. PMID- 15823121 TI - Prognostic importance of vascular endothelial growth factor and its receptors in the uterine sarcoma. AB - Vascular endothelial growth factor (VEGF) and its receptors play an important role in tumor progression; however, there is no report regarding this factor in uterine sarcoma. Thirty-nine patients with uterine sarcoma, 14 carcinosarcomas, 4 endometrial stromal sarcomas, and 21 leiomyosarcomas, were studied. By immunohistochemical staining, VEGF was not detected in normal uterine smooth muscle, but VEGF receptor-1 (flt-1) and VEGF receptor-2 (flk-1) were observed in 14 and 4 of 14 normal smooth muscles, respectively. Of 39 sarcomas, 25 expressed VEGF, and 38 and 34 sarcomas expressed flt-1 and flk-1 at various intensities, respectively. The staining intensity of VEGF, flt-1, and flk-1 was significantly higher in sarcoma than in normal uterine smooth muscle, but that of phospho-flt-1 (p-flt-1) was significantly lower in sarcoma than in normal uterine smooth muscle. When sarcomas were divided into two groups according to staining intensity, a significant difference in survival curves was observed in only p-flt 1 of leiomyosarcoma (P= 0.008), and in all sarcomas, a lower survival curve was also observed in the high staining intensity group than in the low staining intensity group, although there was no significant difference (P= 0.102). In conclusion, VEGF and its receptors are suggested to be involved in progression of uterine sarcoma, but only the p-flt-1 level significantly affected the survival of leiomyosarcoma patients. PMID- 15823122 TI - Utility of MIB-1 and estrogen and progesterone receptor in distinguishing between endometrial stromal sarcomas and endometrial stromal nodules, highly cellular leiomyomas. AB - It is difficult to differentiate between an endometrial stromal nodule (ESN) and endometrial stromal sarcoma (ESS) in curettage specimen, and the recommended therapy of endometrial stromal neoplasm is hysterectomy. If we could discriminate ESS from ESN in curettage specimens, there would be an opportunity to treat ESN by local excision rather than by hysterectomy. We analyzed MIB-1 and estrogen and progesterone receptor (ER/PR) expression in a retrospective series of 8 ESSs, 7 ESNs, and 17 highly cellular leiomyomas obtained from hysterectomy specimens. ESSs expressed MIB-1 more frequently than ESNs (P < 0.05), and ESSs had a tendency to express ER less frequently than ESNs (P= 0.08). We observed that in spite of showing MIB-1 expression to some extent, highly cellular leiomyomas usually could not reach ESSs' level and frequency of MIB-1 expression in the current study. Although MIB-1 and ER appear to be promising markers in the differential diagnosis of ESSs, a larger study would be necessary to confirm their validity. PMID- 15823123 TI - Human telomerase RNA as endogenous control in endometrial tissue. AB - Telomerase is a reverse transcriptase that adds repetitive telomere sequences to the end of chromosomes, which is thought to be essential for cellular immortality and oncogenesis. The enzyme consists of three subunits: human telomerase reverse transcriptase (hTERT), human telomerase RNA (hTR), and telomerase protein 1 (TP1). The hTERT subunit determines the activity of telomerase as an enzyme and is detected in most human tumors and regenerative cells. But many studies have revealed that hTR and TP1 are expressed constitutively. This results suggest that the hTR and TP1 subunits may be potentially good markers of endogenous RNA control. Endometrial dating was determined from the pathomorphology of the endometrium and classified into normal proliferative endometrium, endometrial hyperplasia (simple, complex, and atypical), and endometrial adenocarcinoma. The analysis of the expression of the hTERT, TP1, and hTR telomerase subunits was performed by a quantitative polymerase chain reaction method, based on fluorescent TaqMan methodology (ABI Prism 7,700 Sequence Detection System) capable of measuring fluorescence in real time. The aim of the study was an analysis of the expression profiles of genes encoding hTR and TP1 telomerase subunits in normal endometrium, endometrial hyperplasia, and adenocarcinoma for the estimation of the possibility of once application in endogenous RNA control of gene analysis in the endometrium. The nonparametric Mann-Whitney U test and analysis of variance Friedman test were used to evaluate the variation in telomerase subunit mRNA level between normal endometrium, and endometrial hyperplasia and adenocarcinoma. The results confirm the hTR subunit expression as a good marker of endogenous control in quantitative analysis of gene transcription in endometrial tissue. TP1 subunit transcriptions have not been detected constitutively in our study. PMID- 15823124 TI - Differential regulation of protein kinase C activity by modulating factors and Euonymus alatus (Thunb.) Sieb in human myometrial and uterine leiomyomal smooth muscle cells. AB - Human myometrial smooth muscle cells (MSMC) showed high protein kinase C (PKC) activity when a maximal dose of PKC-activating phorbol ester was used, while uterine leiomyomal cells (ULMC) showed only 6-12% of PKC activity. MSMC exhibited a low proliferation rate, whereas ULMC exhibited a high proliferation rate. These different cell types of MSMC and ULMC responded to 10 U/mL thrombin, with a twofold stimulation of PKC activity. Downregulation of PKC activity was found when MSMC were treated with phorbol ester or with transforming growth factor beta2. We concluded that differences in PKC activity exist between MSMC and ULMC, which may be related to their different proliferative activity. ULMC treated with Euonymus alatus (Thunb.) Sieb (EA), known as "gui-jun woo" in Korea, which is used for leiomyomal tumors, exhibited a much lower proliferation rate than untreated cells, suggesting that EA inhibited the cellular proliferation of ULMC. The PKC activity of MSMC by EA treatment (50 microg/mL) changed little. ULMC showed increased PKC activity by addition of EA, indicating that PKC is activated by EA. The EA-treated ULMC were differentiated into phenotypes characteristic for normal untransformed cells, since the EA-treated cells possess higher PKC activity than untreated cells. PMID- 15823125 TI - Steppe by steppe: gynecological oncology on the Mongolian plains. AB - The objective was to describe the experiences of one gynecologic oncologist in Mongolia. Gynecologic oncologists can contribute to the health of women in developing countries through education of the health care team and research into novel ways to deliver health care. Partnering with our colleagues in developing countries is a personally enriching experience. The development of gynecologic cancer care must ultimately be culturally relevant. PMID- 15823126 TI - Extrauterine mullerian adenosarcoma associated with endometriosis and rectal villotubular adenoma: report of a case and review of the literature. AB - Extrauterine mullerian adenosarcoma is rare and is thought to arise from endometriotic deposits or pluripotent mesothelial and mesenchymal cells of the pelvic cavity. Concomitant colon-rectal neoplasm was reported but the relationship between extrauterine adenosarcoma and colon tumor was not studied. We describe an extrauterine adenosarcoma with a concomitant rectal tubulovillous adenoma. The patient had a long-term history of endometriosis and unopposed estrogen therapy. Immunohistochemical study was performed to investigate the origin and nature of the adenosarcoma. Immunostaining provided the evidence to distinguish between rectal tumor and adenosarcoma. Extrauterine adenosarcoma may arise from endometriosis, and hormone replacement therapy may have some role in the malignant transformation process. Adenosarcoma should be considered in the differential diagnosis of a new pelvic tumor in a patient with a history of endometriosis. PMID- 15823127 TI - Villoglandular adenocarcinoma of the cervix recurrent in an episiotomy scar: a case report in a 32-year-old female. AB - Cervical cancer during pregnancy is rare, occurring in approximately 3% of cervical cancer cases. Considerable controversy exists as to the long-term prognosis of patients diagnosed during pregnancy. A 32-year-old female presented with vaginal spotting in April 1998. A prenatal smear in December 1996 revealed atypical glandular cells of undetermined significance. A sterile speculum exam in April 1997 at 31-week gestational age revealed a polyp on the anterior lip of the cervix, pathology consistent with a well-differentiated villoglandular adenocarcinoma. In August 1997, the patient underwent a radical hysterectomy with pelvic/para-aortic lymphadenectomy. In April 2001, she represented with nodular perineal mass in the episiotomy incision. She received preoperative radiotherapy with a near-complete response and remained without disease for >10 months. It appears that a less radical procedure can offer significant therapeutic value. Preoperative radiotherapy proved effective at achieving a near-complete response. The patient underwent a wide local excision of the perineal area with resultant negative margins. PMID- 15823128 TI - A case with multiple gynecological malignancies. AB - A patient with cervical non-Hodgkin lymphoma was treated with chemotherapy. Fourteen months after the diagnosis of the lymphoma, an endometrial adenocarcinoma was detected as a secondary malignant tumor. The patient was treated with surgery followed by radiotherapy. Approximately 7 years after the diagnosis of endometrial cancer, vaginal invasive squamous cell carcinoma was diagnosed as the third primary malignancy, and a second-line palliative radiotherapy was applied. Seven months after the last radiotherapy, postradiational sarcoma in the vagina was diagnosed. Congenital and acquired immune system disorders, viral oncogenes, and various human leukocyte antigen (HLA) types were investigated. Total blood count and lymphocyte subset analysis were performed, and CD4+ lymphopenia was detected. Serologic tests were carried out for human immunodeficiency virus, hepatitis B virus, human papillomavirus, Epstein-Barr virus, and herpes simplex virus infection. Epstein-Barr virus viral capsid antigen IgG was found positive. Low-risk human papillomavirus panel was detected by Hybrid Capture method in the cervical smear. The HLA investigation revealed HLA-A2, HLA-A3, HLA-B57, HLA-B35, HLA-B4, HLA-B6, HLA-DR3, HLA-DR1, HLA DR51, HLA-DR52, HLA-DQ6(1), and HLA-DQ7(3). The patient died because of the disease. PMID- 15823129 TI - Umbilical metastasis (Sister Joseph's nodule) as a first sign of a disseminated ovarian carcinoma: comparative immunohistochemical analysis of primary tumor and its metastases. AB - The case of a 46-year-old female with umbilical metastasis as a first sign of an ovarian carcinoma is reported with the results of immunohistochemical analysis of primary tumor and lymph node and umbilical metastases. All specimens were positive for cytokeratin 7, CA 125, E-cadherin, alpha-, beta-, and gamma-catenin, as well as for MSH2. Staining with cytokeratin 20 and MLH1 was negative, and Ki 67 labeled from 5% (in the center of the lesions) to over 25% (at the periphery of the lesions) of the nuclei. Beta-catenin showed membranous positivity in the central parts and absence of staining at the periphery of ovarian tumor and umbilical metastasis, whereas lymph node metastasis presented with uniform reaction throughout. The results of immunohistochemical staining could point to the mechanisms employed by malignant tumors during invasion and growth of metastasis and suggest the possible role of the microenvironment in the expression of some adhesion molecules on tumor cells. PMID- 15823130 TI - Immature ovarian teratoma in a young girl: very short course and lethal outcome. A case report. AB - Immature teratomas of the ovary represent less than 1% of all ovarian teratomas. They contain several tissues that derive from the three embryologic layers: ectoderm, mesoderm, and endoderm. We report the case of a 17-year-old girl who presented with an immature teratoma of the left ovary that recurred 3 months later as an inoperable malignant neuroepithelial peritoneal tumor resembling a glioblastoma. She died 3 weeks after the second explorative operation. The very aggressive course and fast lethal outcome could be explained by the patient's refusal to receive chemotherapy after the initial operation. PMID- 15823131 TI - Placental site trophoblastic tumor presenting as subaponeurotic metastasis. AB - Cases of metastatic placental site trophoblastic tumor (PSTT) have a very poor prognosis because these tumors tend to be less sensitive to chemotherapy than other types of gestational trophoblastic disease. We describe the case of a 25 year-old woman who presented with occipital tumor and abnormal vaginal bleeding. Hysterectomy, bilateral salpingo-oophorectomy, and occipital tumor removal revealed a primary PSTT in the uterus, with ovarian and occipital subaponeurotic metastases. She received etoposide, methotrexate, actinomycin-D/cyclophosphamide, vincristine chemotherapy and had a complete clinical remission. Fifteen months later, she had a recurrent subaponeurotic occipital tumor invading the cranium and underwent tumor removal along with cranial bone followed by local irradiation. She was then treated with etoposide, cis-platinum/etoposide, methotrexate, actinomycin-D chemotherapy and again had a remission for 5 months. The patient, however, had a left parietal subaponeurotic tumor, invading the dura mater, and received local irradiation. Soon after, she developed left orbital bone metastasis, treated by local irradiation. These bone metastases responded to the radiation completely. However, multiple organ metastases were found, and she died of the disease. This represents the first case of PSTT with initial subaponeurotic metastasis in a living patient. New modalities of treatment for high-risk or metastatic PSTT need to be developed. PMID- 15823132 TI - A systematic nerve-sparing radical hysterectomy technique in invasive cervical cancer for preserving postsurgical bladder function. AB - The objective of this study is to describe a technique for preserving the autonomic nerve systematically, including the hypogastric nerves, pelvic splanchnic nerves, and pelvic plexus and its vesical branches, based on anatomic considerations for the autonomic nerves innervating the urinary bladder, in radical hysterectomies and to assess postsurgical bladder function. A nerve sparing radical hysterectomy was carried out on 27 consecutive patients with uterine cervical cancer treated between 2000 and 2002. The FIGO stages of the disease consisted of 10 stage Ib1, 6 stage Ib2, 3 stage IIa, and 8 stage IIb. The nerve-sparing procedure was successfully completed in 22 of the 27 patients (81.5%) in the study. At 1 year after the operation, bladder symptoms were significantly improved in the nerve-sparing group compared to the non-nerve sparing group. Urinary incontinence and abnormal (diminished) bladder sensation were observed in three of the five patients (two patients had both symptoms), for whom the nerve-sparing procedure could not be performed, but none of the 22 patients for whom the nerve-sparing procedure was performed had incontinence, and only two patients had abnormal (increased) bladder sensation (P= 0.0034 for incontinence and P= 0.030 for abnormal bladder sensation). The patients' survival was not adversely affected by the nerve-sparing procedure. Although it is still preliminary, the surgical technique described in this report is thought to be effective for preserving bladder function, and thus, the quality of life could be improved for patients with cervical cancer who are treated with a radical hysterectomy. For further evaluation of the efficacy of nerve-sparing radical hysterectomy, a prospective randomized trial needs to be performed. PMID- 15823154 TI - Changes in mental disorder distribution among suicide attempters in mid-west area of Kanagawa. AB - To clarify changes in mental disorder distribution in suicide attempters, a comparison was made between the period when an economic recession started (1992, 1993) and the period when the recession became serious (2000). The subjects were 212 suicide attempters admitted to the Emergency Medical Center of Tokai University Hospital during a 2-year period between 1992 and 1993 (group A) and 255 suicide attempters admitted during a 1-year period between January and December 2000 (group B). Mental disorders were classified according to the International Classification of Diseases (10th revision; ICD-10). Distribution was compared between groups A and B and significant differences were observed in mental disorder distribution (P < 0.001). The percentages of subjects with depression, those with neurotic disorders, and those with other disorders were higher in group B than in group A. Conversely, the percentage of subjects with schizophrenia was lower in group B than in group A. Mental disorder distribution in suicide attempters significantly changed. However, it was not certain that this change was related to the economic recession and increased unemployment in Japan. PMID- 15823155 TI - Prediction of first episode of panic attack among white-collar workers. AB - The purpose of the present study was to elucidate a longitudinal matrix of the etiology for first-episode panic attack among white-collar workers. A path model was designed for this purpose. A 5-year, open-cohort study was carried out in a Japanese company. To evaluate the risk factors associated with the onset of a first episode of panic attack, the odds ratios of a new episode of panic attack were calculated by logistic regression. The path model contained five predictor variables: gender difference, overprotection, neuroticism, lifetime history of major depression, and recent stressful life events. The logistic regression analysis indicated that a person with a lifetime history of major depression and recent stressful life events had a fivefold and a threefold higher risk of panic attacks at follow up, respectively. The path model for the prediction of a first episode of panic attack fitted the data well. However, this model presented low accountability for the variance in the ultimate dependent variables, the first episode of panic attack. Three predictors (neuroticism, lifetime history of major depression, and recent stressful life events) had a direct effect on the risk for a first episode of panic attack, whereas gender difference and overprotection had no direct effect. The present model could not fully predict first episodes of panic attack in white-collar workers. To make a path model for the prediction of the first episode of panic attack, other strong predictor variables, which were not surveyed in the present study, are needed. It is suggested that genetic variables are among the other strong predictor variables. A new path model containing genetic variables (e.g. family history etc.) will be needed to predict the first episode of panic attack. PMID- 15823156 TI - Alexithymia and dissociative tendencies in an adolescent sample from Eastern Turkey. AB - Alexithymia and dissociative reactions are two strategies that have been put forward as coping mechanisms to alleviate painful emotions. Adult studies reveal an association between alexithymia and dissociation. In line with the coping hypothesis, it was predicted that the relationship between alexithymia and dissociative tendencies would be partly mediated by current levels of stress and past traumatic experiences. Dissociation may also be related to enhanced fantasizing, although alexithymia has traditionally been associated with an incapacity to fantasize. This relationship has not been studied well in adolescents. In the present study, 173 randomly selected high school students from Eastern Turkey were assessed with the Adolescent Dissociative Experiences Scale, the Beck Depression and Anxiety inventories, the Rosenberg Self-esteem Scale and the Toronto Alexithymia Scale. Correlation analyses followed by stepwise regression analyses were performed. "Difficulty identifying feelings" subscale of Toronto Alexithymia Scale, anxiety level and the history of physical abuse emerged as predictors of dissociative tendencies. It seems that in this group, dissociation is associated with anxiety and with difficulty in identifying feelings. Physical abuse also contributes to dissociation. The adolescent who is alexithymic may be more prone to dissociation when compared to their non alexithymic peers. Helping alexithymic adolescents describe their body experiences may decrease their tendency to dissociation by increasing their ability to verbally identify their stressors. PMID- 15823157 TI - Validation of Impact of Events Scale in nurses under threat of contagion by severe acute respiratory syndrome. AB - The purpose of the present study was to validate the Impact of Events Scale (IES) in a sample of nurses working under threat of severe acute respiratory syndrome (SARS). The internal consistency, construct validity and convergent validity of the instrument were examined in a sample of 128 nurses during the SARS outbreak in Taiwan. Principal component analysis followed by a quartimax rotation were used to derive a two-factor solution, labeled intrusion (factor 1) and avoidance (factor 2), with both factors accounting for 50.7% of the explained variance. The total Cronbach's alpha of 0.90 reflected the good internal consistency of the instrument. Correlation with the Symptoms Checklist 90-R demonstrated the convergent validity of the IES. In conclusion, the IES can be used as a convenient, reliable and valid instrument for evaluation of the psychological distress of nurses working with the highly contagious disease. PMID- 15823158 TI - Association between the 5-HT6 receptor C267T polymorphism and response to antidepressant treatment in major depressive disorder. AB - The purpose of the present study was to determine if a 5-HT6 receptor polymorphism is associated with antidepressant treatment response in major depressive disorder (MDD). Ninety-one patients with MDD, compared with 127 normal control subjects, were evaluated after an 8-week treatment period. An association analysis revealed no differences in genotype and allele distribution between patients with MDD and normal control subjects. However, there were significant differences in the treatment response in some Hamilton Depression Rating Scale (HAM-D) scores (sleep, activity, somatic anxiety, and total) between genotypes. Moreover, the heterozygote group (CT genotype) had significantly better treatment response than the homozygote group (CC + TT genotypes), especially in the somatic anxiety subcategory and the total score of HAM-D. These findings imply that a 5 HT6 receptor polymorphism (C267T) is associated with treatment response in MDD. PMID- 15823159 TI - Predicting features of later development of schizophrenia among patients with acute and transient psychotic disorder. AB - It is acknowledged that a considerable proportion of patients who have an episode of acute and transient psychotic disorder (ATPD), will develop schizophrenia after the episode is remitted. If features discriminating the patients who later developed schizophrenia from those who underwent a monomorphous ATPD-only course are discerned, they may be useful for predicting the prognosis. The authors performed a chart review study of those who were hospitalized with the diagnosis of ATPD and followed up thereafter for >5 years and compared two subject groups: 10 later-schizophrenia-developing (SD) patients and 15 non-schizophrenia developing (non-SD) patients. Assessment of the subjects, conducted by two psychiatrist raters blind to the subject groups, included frequency of ATPD episodes, adaptation levels and life events before admission, and psychiatric symptoms before and after admission. The interrater reliability of the assessment proved to be favorable. As a result, early recurrence of psychotic episodes in the follow-up period, lack of acute upsurge of insomnia in an early phase of the episode and poor premorbid heterosexual relations characterized the SD patients. The result indicated heterogeneity of ATPD episodes. PMID- 15823160 TI - Substance abuse, conduct disorder and crime: assessment in a juvenile detention house in Istanbul, Turkey. AB - The purpose of the present study was to determine the rate of substance abuse in the juvenile detention house and to determine the relationship between crime and substance abuse and conduct disorder. Two hundred and thirty cases in the biggest juvenile detention house in Istanbul, Turkey were assessed according to Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) criteria. Law files and data of crime were examined. A total of 80 out of 230 juvenile detainees (34.8%) were found to have substance abuse excluding nicotine and alcohol. The substances abused in preferential order were cannabis (72.5%), volatile substances (21.3% bally and 3.7% thinner; 25%) and sedative hypnotic drugs and biperidents (2.5%). The rate of conduct disorder was 46.3% in substance abusers and 25.3% in the others (odds ratio: 2.536). The rate of substance abuse was 48.5% in the juveniles who had committed multiple crimes and 14.1% in the others (odds ratio: 5.735). The study shows that conduct disorder was very high in juvenile detainees. Conduct disorder was higher in substance-abusing than in non-abusing juvenile detainees. Substance-abusing juvenile detainees were found to have a higher detention rate than non-abusing juvenile detainees. There was a close relation between conduct disorder and substance abuse and multiple crimes. In the light of these results, diagnosis and treatment for conduct disorder in juvenile detainees are of great importance. PMID- 15823161 TI - Neural basis of fear conditioning induced by video clip: positron emission tomography study. AB - In patients with post-traumatic stress disorder (PTSD), re-experiencing the trauma is often induced by external cues in the environment. The cues, which were emotionally neutral for the patients before the traumatic event, become fearful ones after the event. This phenomenon is considered to be associated with fear conditioning. The paradigm was set up so that the emotionality changes in the patients with PTSD would be reproduced, and the regional cerebral blood flow (rCBF) measured with positron emission tomography (PET) was compared during exposure to the same stimuli before and after acquisition of fear conditioning. Ten healthy male subjects were asked to look at some emotionally neutral photos, then to watch a video with fearful content that also contained images similar to that presented in the photos, and afterwards to look at the photos again. Five of the 10 subjects felt that the object in the photos was more fearful after watching the video than before, and they were considered to have acquired fear conditioning. In those five subjects, the rCBF in the right amygdala and the left posterior cingulate gyrus after acquisition of fear conditioning significantly increased relative to the rCBF before conditioning. Thus, these regions seem to have a critical role in fear conditioning. PMID- 15823162 TI - Quality of life of schizophrenic patients living in the community: the relationships with personal characteristics, objective indicators and self esteem. AB - The relationship of personal characteristics, objective indicators, and self esteem to quality of life (QOL) was investigated cross-sectionally in 73 Japanese schizophrenic patients, and the question of how objective conditions affect subjective evaluation was evaluated. Depressive mood and uncooperativeness were negatively correlated with self-esteem, and self-esteem was positively correlated with QOL. Self-esteem was considered to directly affect QOL, and depressive mood and uncooperativeness to affect QOL via self-esteem. Findings suggest that, in order to improve the QOL of schizophrenia patients, interventions to help alleviate psychiatric symptoms (e.g. depressive mood, uncooperativeness) must be utilized. A change in Japanese society itself must occur so that the consequences of stigma may be reduced and patient coping skills improved. PMID- 15823163 TI - Norms of performance of sustained attention among a community sample: Continuous Performance Test study. AB - The Continuous Performance Test (CPT) is a widely used measure of sustained attention, which is a preferred tool for assessing various mental functions. A well-established norm for CPT is essential when choosing a suitable threshold for classifying individuals as either case (CPT impairment) or non-case. The CPT performance of 900 adults who were randomly chosen from a community survey was measured to establish the norms for subgroups with different gender, age, and educational levels. The results revealed that age and educational level are significantly associated with the performance sensitivity (d') of CPT. Male subjects perform better than female subjects. Seventeen percent of the subjects scored higher on the masked CPT than on the unmasked CPT. Subjects who could not finish the masked CPT had the characteristics of older age and fewer years of education. When classifying a patient as case or non-case, his/her CPT performance should be considered relative to the norms for his/her gender, age, and educational levels. PMID- 15823164 TI - Factors associated with the development of panic attack and panic disorder: survey in the Japanese population. AB - Environmental factors, in addition to genetic factors, may be related to the development of panic attack (PA) and panic disorder (PD). Previous studies suggested that there may be seasonal variation in the onset of PA/PD and possibly a higher prevalence of PA/PD in colder areas. Also observed were lactate-induced PA and elevated serum cholesterol in PD patients. These suggest that living environment and lifestyle, such as weather conditions, preference of food and physical exercise, might play a role in the occurrence of PA and PD. The present study explored the association of such candidate factors with the development of PA and PD in 4000 Japanese subjects, using a questionnaire. The subjects were recruited from the general population of Japan, using stratified random sampling. Logistic regression with stepwise selection of variables was employed for statistical analysis. Variables including "dislike of physical exercise", mostly in female subjects, and "living in areas with longer winter", in male subjects, were suggested for associations with PA and PD among the candidate factors. The result is preliminary but indicates that lifestyle such as like/dislike of physical exercise and environmental factors including weather conditions could play a partial role in the development of PA and PD. Further investigations are required before firm conclusions can be reached. PMID- 15823165 TI - Antipsychotics and dopamine transporter gene polymorphisms in delirium patients. AB - The main objective of the present study was to determine the relationship between treatment responses of delirium and genetic polymorphisms in the dopamine transporter. The optimal dosages of haloperidol and risperidone in the treatment of delirium were also investigated. Either haloperidol or risperidone was administered to delirium patients, and delirium symptoms were measured daily until remission. Variable number of tandem repeat (VNTR) polymorphisms of the dopamine transporter were determined using the polymerase chain reaction. Among 42 subjects, symptoms of delirium appeared a mean of 9.68 days after hospitalization. A majority of the subjects (83.3%) had the type 10/10 polymorphism. Dosages of haloperidol and risperidone at the day of recovery were 1.67 mg/day (SD = 1.32; range 0.5-2.5 mg/day) and 1.19 mg/day (SD = 1.14; range 0.5-5.0 mg/day), respectively. The mean drug response time was 8.5 days in the haloperidol group and 4.8 days in the risperidone group (no significant difference). The response rates at the 3rd and 7th days after medication did not differ with either the drug group or the dopamine transporter polymorphism. Relatively low doses of risperidone and haloperidol exhibited similar efficacies, and dopamine transporter polymorphisms do not appear to play a major role in the action of antipsychotics on delirium. PMID- 15823166 TI - Serum leptin levels in obsessive-compulsive disorder. AB - The aim of the present study was to evaluate serum leptin levels to demonstrate whether or not its eventual alterations might have an etiopathogenetic significance in patients with obsessive-compulsive disorder (OCD). Thus, it was planned to examine whether serum leptin levels were affected by pure OCD (OCD-D), pure depression (D) or the comorbidity of OCD and depression (OCD+D). Forty-four patients with OCD (27 with OCD-D and 17 with OCD+D), 38 depressed patients and 30 control subjects were enrolled and serum leptin and cortisol levels were measured. According to the mean leptin levels, no significant difference was found between the OCD-D and control groups and between the OCD+D and D groups, while statistically significantly lower levels were found in the OCD+D and D groups than in control group. Significant difference in the mean leptin levels was found among groups even after controlling for body mass index or sex. The present study confirms the strong relationship between serum leptin and cortisol values and suggests that reduced leptin levels, rather than having an etiopathogenetic significance in patients with OCD, seem to be associated with patients with OCD and depression but not with pure OCD patients, and that OCD may be a heterogeneous subtype containing some biological indications of anxiety and affective disorders. PMID- 15823167 TI - Difference in the characteristics of subjective and objective sleepiness between narcolepsy and essential hypersomnia. AB - The present study was conducted to investigate the difference in the characteristics of daytime sleepiness between narcolepsy and essential hypersomnia and to identify the relationship between the Epworth Sleepiness Scale (ESS) and the Multiple Sleep Latency Test (MSLT) in patients with these two disorders. Subjects consisted of 34 patients with essential hypersomnia (32.4 +/- 11.0 years old), 52 patients with narcolepsy (29.0 +/- 13.8 years old), and 45 control subjects (33.3 +/- 6.6 years old). The subjects completed the ESS and underwent MSLT following a regular sleep-wake schedule for over 2 weeks. The ESS scores were pathologically high and mean sleep latency on MSLT was short, not only in narcolepsy but also in essential hypersomnia. With respect to sleep latencies on each MSLT session, both essential hypersomnia and control subjects had the smallest value at 14:00, while narcolepsy lacked any statistical change at this time period. The correlation between ESS and mean sleep latency on MSLT was higher in essential hypersomnia than in narcolepsy, and the correlation was strongest for the session performed at 14:00. Based on the ESS and MSLT results, the severity of excessive daytime sleepiness was significantly milder in essential hypersomnia compared with that in narcolepsy. The results also indicate that diurnal variation of sleepiness was maintained, and the correlation between subjective and objective sleepiness was relatively maintained in essential hypersomnia compared to narcolepsy. It is suggested that the mild disease severity of essential hypersomnia contributed to the formation of these characteristics. PMID- 15823168 TI - Comparison of childhood disintegrative disorder and disintegrative psychosis not diagnosed as childhood disintegrative disorder. AB - To clarify the difference of Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) childhood disintegrative disorder (CDD) from International Classification of Diseases (9th revision; ICD-9) disintegrative psychosis (DP), 10 CDD children (mean age, 7.7 years) and 18 DP children (mean, 6.5 years) not diagnosed as CDD divided into DSM-IV autistic disorder (DP-AD; n = 11) and pervasive developmental disorders not otherwise specified (DP-PDDNOS; n = 7) were compared on 31 variables not directly related to the normalcy before regression. The CDD, DP-AD, and DP-PDDNOS groups did not differ significantly in 28 variables. The DP-PDDNOS group met significantly a smaller number of items in criterion A of DSM-IV autistic disorder criteria than the CDD and DP-AD groups, both of which did not differ significantly in this respect. The CDD group tended to be more abnormal in auditory responsiveness and verbal communication than the DP-PDDNOS group. While CDD is distinct from DP-PDDNOS, its validity apart from AD with regression remains to be studied. PMID- 15823169 TI - Actigraphic assessment of sleep in young adults with Asperger syndrome. AB - A high prevalence of subjective insomnia in adults with Asperger syndrome has been reported. In the present study the sleep quality of these patients was studied using wrist actigraphy. Nineteen adults with Asperger syndrome and frequent feelings of insomnia were compared with 10 controls devoid of neuropsychiatric disorders and subjective sleep problems during six consecutive nights. The patients had similar actigraphic sleep profile to the controls. The subjective low sleep quality in patients was not reflected in actigraphic assessment of sleep. This finding suggests that pervasive anxiety inherent in persons with Asperger syndrome predisposes them to insomnia complaints. PMID- 15823170 TI - Influence of duration of untreated psychosis on auditory P300 in drug-naive and first-episode schizophrenia. AB - P300 amplitude reduction in schizophrenia is, according to previous studies, partially recovered by treatment with neuroleptics. However, whether this medication-induced P300 recovery is associated with duration of untreated psychosis (DUP) remains unreported; the present study is a preliminary examination of this question. Auditory P300 was recorded from 18 drug-naive and first-episode schizophrenia patients, among whom 10 were identified as short DUP, and eight as long DUP. Follow-up event-related potential tests were carried out after treatment with haloperidol or bromperidol for approximately 2 months. Recovery of P300 amplitude was replicated after neuroleptic medication was administered. A significant interaction was found between DUP and the medication effect in P300 amplitude over the left temporo-parietal area; a significant P300 recovery was seen in short DUP but not in long DUP. These results suggest that first-episode schizophrenia patients with long DUP might have severe impairments in the left temporal structures, supporting DUP as a key variable in future neurobiological studies of first-episode schizophrenia. PMID- 15823171 TI - Selective memory impairment for personally familiar colors following encephalitis. AB - A 19-year-old man presented with a peculiar memory impairment following non herpetic viral encephalitis. He was exclusively impaired in generating images for colors of possessions previously well known to him, and consequently these seemed less familiar. He appeared unable to access personally familiar and autobiographical color attributes. Single-photon emission computed tomography and electroencephalogram findings suggest that the right-sided functional abnormalities, specifically in the frontal and temporal regions, were implicated as the functional locus for the patient's unique problem. PMID- 15823172 TI - Obsessive-compulsive symptoms in clozapine-treated schizophrenic patients. AB - The aim of the present study was to assess the occurrence of obsessive-compulsive symptoms (OCS) in schizophrenic patients treated with clozapine, and to examine the relationship between OCS and other clinical variables. The results support earlier findings which suggest that clozapine produces or unmasks OCS. In addition, the severity of OCS was not related to other dimensions of psychopathology, severity of illness, clinical improvement or dose and duration of clozapine treatment. PMID- 15823173 TI - Consecutive monitoring of sleep disturbance for four nights at the top of Mt Fuji (3776 m). AB - The purpose of the present study was to conduct consecutive monitoring of sleep from the second to the fifth night at altitude. Three healthy male subjects climbed the top of Mt Fuji (3776 m) and stayed there for 6 days. Polysomnographic recordings were performed during this period and control recordings were made at sea level 1 month after the mountaineering. Disturbed sleep characterized by an increased number of arousals and/or long wake time was observed to persist through the fifth night in all subjects. These results suggest that sleep disturbance might persist during initial days at altitude despite cumulating sleep pressure due to poor night's sleep. PMID- 15823174 TI - Incidence of note-leaving remains constant despite increasing suicide rates. AB - Suicide notes (SN) are potentially valuable sources of information about the psychological states of the suicidal person. It was hypothesized that there was a significant relation between suicide rate and note-leaving rate and that the incidence of note-leaving was increased during prolonged economic recession. During 21 years (1981-2001) in Kobe, of a total of 18 558 violent deaths, 5161 were due to suicide (27.8%), with 3417 male cases (66.2%) and 1754 female cases (33.8%). For each year the annual suicide rates and note-leaving rates were calculated, and this represents the percentage of committed suicides in which SN were left, among all suicide victims. In spite of the prolonged economic slump, the note-leaving rate remained almost constant (23.4-36.2%). Pearson's correlation coefficient showed no significant correlation between suicide rate and note-leaving rates (r = 0.27, P = 0.23). The finding that the incidence of note-leaving remains constant despite increasing suicide rates may suggest that the reasons for suicide do not affect note-leaving. There are cross-cultural, ethnic, and racial variations in suicidal behaviors. Although this finding may be specific in Japan, further studies of SN are needed to help clarify the suicidal states of mind. PMID- 15823176 TI - Thoracic embolotherapy for life-threatening haemoptysis: a pulmonologist's perspective. AB - Haemoptysis is the expectoration of blood from the respiratory tract and is life threatening when blood obstructs the major airway causing asphyxiation. This may occur in the presence or absence of pre-existing compromised pulmonary reserve and medical comorbidities. Life-threatening haemoptysis is a relatively common clinical situation encountered by pulmonologists and thoracic embolotherapy is a key therapeutic option. Although thoracic embolotherapy primarily involves bronchial arterial embolization, it also encompasses embolization of non bronchial systemic arteries and the pulmonary arterial supply. A pulmonologist's perspective on this topic is presented. Pertinent areas related to this theme, namely surgery and the role of bronchoscopy in life-threatening haemoptysis are discussed briefly. PMID- 15823177 TI - Management of tuberculous pleuritis: can we do better? AB - Management of patients with tuberculous pleuritis can be improved by establishing early diagnosis accurately, administering effective chemotherapy, and close monitoring of progress for early detection and prompt management of severe pleural inflammation in the hope of preventing or reducing subsequent residual pleural fibrosis. In addition to the conventional diagnostic tools, chemical markers, especially pleural fluid adenosine deaminase and interferon-gamma levels and new microbiological tests such as polymerase chain reaction and BACTEC culture of pleural biopsy specimens for Mycobacterium tuberculosis, can increase the diagnostic yield for tuberculous pleuritis. Indicators of the severity of pleural inflammation, including high pleural fluid tumour necrosis factor-alpha and lysozyme levels, and low pleural fluid glucose and pH, can help to predict residual pleural fibrosis. It is likely that patients will require surgery: (i) complete drainage of pleural fluid for prevention; and (ii) pleurectomy for the treatment of residual pleural fibrosis. PMID- 15823178 TI - Augmentation of allergic inflammation in the airways of cyclooxygenase-2 deficient mice. AB - OBJECTIVE: Airway cyclooxygenase-2 (COX-2) is induced by cytokine-mediated inflammation such as occurs in asthma. However, the role of COX-2 in the pathophysiology of asthma is not fully understood. METHODS: Allergic inflammation, airway responsiveness to methacholine and mucous cell metaplasia after ovalbumin sensitization in the airways of COX-2 deficient (-/-) mice, COX-2 (+/+) mice and C57BL/6J mice treated with a selective COX-2 inhibitor, nimesulide were assessed. Histology, cell analysis, measurements of arachidonic acid metabolites and Th2 cytokine levels in bronchoalveolar lavage fluid (BALF), and measurement of serum IgE were performed. RESULTS: Eosinophil infiltration into the airway wall, and the number of eosinophils in BALF were greater in sensitized COX-2 (-/-) mice than in sensitized COX-2 (+/+) mice. The levels of cysteinyl leukotrienes (LTC4/D4/E4), prostaglandin E2 (PGE2) and interleukin (IL)-13 as well as airway responsiveness did not differ in COX-2 (-/-) mice and COX-2 (+/+) mice. However, sensitized COX-2 (-/-) mice had higher LTC4/D4/E4 and lower PGE2 concentrations compared with non-sensitized COX-2 (-/-) mice. The number of PAS/alcian blue-positive airway epithelial cells and serum IgE were elevated in COX-2 (-/-) mice. Nimesulide-treated mice showed augmented eosinophilic inflammation, LTC4/D4/E4 concentrations and mucous cell metaplasia. CONCLUSION: These data indicate that COX-2 deficiency augments allergic inflammation and mucous cell metaplasia. PMID- 15823179 TI - The effect of N-acetylcysteine on Clara cells and Clara cell 16 kDa protein in a murine model of allergen-induced airway inflammation. AB - OBJECTIVE: The aim of this study was to investigate the number of Clara cells and the production and secretion of Clara cell 16 kDa protein (CC16) in a murine model of allergen-induced airway inflammation, as well as the effects of N acetylcysteine (NAC) on CC16 and Clara cell numbers, in order to determine the mechanism of the anti-inflammatory effect of NAC. METHODOLOGY: BALB/c mice were divided into control, ovalbumin (OVA) and NAC groups. An allergen-induced airway inflammation model (OVA group) was established by sensitizing and challenging mice with OVA. NAC was administered as an oral treatment. The number of Clara cells and the production of CC16 were determined by immunohistochemistry. The CC16 levels in bronchoalveolar lavage fluid (BALF) were determined by Western blotting. RESULTS: The proportion of Clara cells in terminal and respiratory bronchioles significantly decreased in the OVA group compared to the control group (P < 0.01). NAC treatment did not change the proportion of Clara cells in the OVA group (P > 0.05). CC16 production by Clara cells in the OVA groups was significantly lower than that of the control group (P < 0.01), but was elevated following NAC treatment (P < 0.05). The CC16 level in BALF of the OVA group was lower than that of the control group (P < 0.01), but was elevated by NAC treatment (P < 0.05). NAC reduced the total number of white cells and the percentage of eosinophils in BALF. Moreover, it inhibited airway inflammation. CONCLUSIONS: The number of Clara cells and the production and secretion of CC16 were reduced in a murine model of allergen-induced airway inflammation. Antioxidants can enhance the expression of CC16, which might be a mechanism by which they suppress airway inflammation. PMID- 15823180 TI - Metabolic activity in skeletal muscles of patients with non-hypoxaemic chronic obstructive pulmonary disease studied by 31P-magnetic resonance spectroscopy. AB - OBJECTIVE: An alteration of high energy phosphate metabolism in muscles may contribute to exercise intolerance. The objective of this study was to clarify the changes in high energy phosphate metabolites in muscles during exercise in patients with non-hypoxaemic chronic obstructive pulmonary disease (COPD), which influences the impairment of muscle metabolism. METHODOLOGY: Calf muscle energy metabolism was studied in eight stable non-hypoxaemic COPD patients and eight control subjects, using 31P-magnetic resonance spectroscopy (MRS). MRS spectra were acquired at rest, during exercise at two levels of intensity, and during recovery. The control subjects exercised under both normoxic and hypoxic conditions. The intensity of exercise was standardized by the maximal isometric voluntary contraction (MVC) of the calf muscle and the cross-sectional area (CSA) of calf muscle. RESULTS: MVC and CSA were lower in COPD patients. No significant differences in intracellular pH, inorganic phosphate/phosphocreatine ratio or percentage recovery in inorganic phosphate/phosphocreatine ratio were observed between the two groups in muscles at rest, during exercise or during recovery. CONCLUSIONS: Muscle metabolites, during exercise standardized by muscle CSA and MVC, did not differ between non-hypoxaemic COPD patients and control subjects. MVC, CSA or both, are assumed to be closely related to muscle metabolism, as no difference in high energy phosphate metabolites was observed for COPD patients compared to control subjects when the load was standardized for MVC and CSA. This suggests that high energy metabolites are consumed to a similar extent in the same muscle volume in non-hypoxaemic COPD patients and control subjects. PMID- 15823181 TI - Zidovudine-induced diaphragmatic contractile dysfunction: impact of an antioxidant diet. AB - OBJECTIVE: Zidovudine (AZT) is a primary drug therapy used to treat HIV-infected individuals. While AZT inhibits replication of HIV, it also induces a drug specific myopathy resulting in altered muscle mitochondria, increased oxidative stress and muscle contractile dysfunction. The purpose of this study was to assess the impact of an antioxidant diet (high in vitamins C and E) on AZT mediated diaphragmatic contractile dysfunction in rodents. METHODOLOGY: Adult, Sprague-Dawley rats were assigned to feeding groups: control (CON, n = 9), AZT treatment (AZT, n = 8), antioxidant diet only (Anti-Ox, n = 6), and AZT + antioxidant diet (AZT + Anti, n = 9). Two costal diaphragm strips were removed from each animal (under surgical anaesthesia) and evaluated for force-frequency relationship, maximal specific tension, and fatigue resistance using an in vitro preparation. RESULTS: Results indicate significant reductions in normalized force production (20-200 Hz), including maximal specific tension, between AZT animals and all other groups. While AZT reduced diaphragm contractility, the addition of an antioxidant diet eliminated this decrease. CONCLUSION: These data suggest that an increase in oxidative stress mediated by AZT may contribute to AZT-induced muscle contractile dysfunction, and that antioxidant vitamin supplementation may help ameliorate this effect. PMID- 15823182 TI - Nasal wash as an alternative to bronchoalveolar lavage in detecting early pulmonary inflammation in children with cystic fibrosis. AB - OBJECTIVE: The aim of this study was to determine whether nasal inflammation reflects pulmonary inflammation in young children with cystic fibrosis (CF), as assessed by inflammatory markers in nasal wash (NW) and bronchoalveolar lavage (BAL), respectively. METHODS: CF patients younger than 6 years of age who were to undergo bronchoscopy for routine BAL from May 2000 to October 2001 were recruited for this study. NW was collected immediately after the patient was sedated for bronchoscopy. Total cell counts (TCC), differential cell counts and interleukin (IL)-8 levels (enzyme-linked immunosorbent assay) were assessed in NW and BAL. RESULTS: In total, 19 children with CF (mean age, 1.9 years; SD, 1.7 years) were included in the study. There was a significant relationship between IL-8 and the percentages of neutrophils in NW (r (2) = 0.76; P < 0.001) and in BAL fluid (r 2 = 0.62; P = 0.006). Similarly, IL-8 concentrations in the NW correlated with those in the BAL (r 2 = 0.48; P = 0.036) and neutrophil percentages in NW correlated significantly with those in BAL (r 2 = 0.7; P = 0.004). CONCLUSION: When measured under 'ideal' conditions, nasal IL-8 reflects lower airway levels and may reflect the inflammatory stimulus that results in neutrophilic inflammation. These data encourage further assessment of nasal wash under clinically appropriate conditions to determine its utility for assessing inflammation in young children with CF. PMID- 15823183 TI - Fluticasone given once versus twice a day: meta-analysis. AB - OBJECTIVE: The aim of this study was to examine the efficacy of fluticasone administered once daily compared to twice daily in asthma. METHODOLOGY: A meta analysis was performed of randomized double-blind trials of at least 4 weeks duration that compared fluticasone administered once versus twice a day and presented data on at least one clinical outcome measure. RESULTS: Six studies of 1517 children and adults with asthma met the inclusion criteria. Studies were predominantly in subjects with moderate asthma, treated with doses of fluticasone ranging from 200 to 500 microg per day. Twice-daily dosing was associated with significantly greater efficacy compared with once-daily dosing, for all outcome measures except night wakenings. The mean (95% CI) differences between twice and once-daily administration for FEV(1) and peak expiratory flow were 0.11 L (0.07 0.16) and 12.9 L/min (8.6-17.1), respectively. Twice-daily fluticasone was associated with significantly fewer withdrawals due to asthma than once-daily fluticasone, with an odds ratio of 0.44 (0.30-0.67). CONCLUSION: The findings suggest that twice-daily administration of fluticasone will provide greater therapeutic benefit than a once-daily morning regimen. PMID- 15823184 TI - Comparison of inhaled corticosteroid combined with theophylline and double-dose inhaled corticosteroid in moderate to severe asthma. AB - OBJECTIVE: Recent studies have found that theophylline exerts anti-inflammatory and immunomodulatory effects. This study was performed to compare the efficacy of inhaled corticosteroids (ICS) combined with slow-release theophylline (SRT) with that of double-dose ICS in asthma control, anti-inflammatory activity and safety. METHODOLOGY: In a randomized, open, parallel, control trial, 41 patients with asthma were randomly treated with either beclomethasone dipropionate 500 microg b.i.d. (BDP group) or a combination of BDP 250 microg b.i.d and SRT 0.2 g b.i.d. (SRT/BDP group) for 6 weeks. At the start and at the end of treatment, lung function testing and sputum induction were performed, and plasma cortisol levels were measured. Sputum was analyzed for cell differential counts and the interleukin (IL)-5 level. Patients kept a record of peak expiratory flow (PEF), symptom score, and beta2-agonist use. RESULTS: Significant increases in the morning and the evening PEF and FEV1 were observed (P < 0.05), together with an obvious reduction in symptom score and beta2-agonist use (P < 0.01). Significant decreases in the percentage eosinophils and IL-5 level in induced sputum also occurred (P < 0.05). However, there was no difference between the two groups for all these parameters. There was no significant change in the plasma cortisol level for either group. CONCLUSIONS: Both ICS combined with SRT and double-dose ICS had the same effect on asthma control, improving symptoms and ameliorating lung function. Both therapies had similar anti-airway inflammatory effects and therapeutic safety. Combining SRT with ICS may allow a reduction in ICS dose when treating asthma. PMID- 15823185 TI - Characteristics of patients with delayed diagnosis of infectious pulmonary tuberculosis. AB - OBJECTIVE: The aim of this study was to identify patient and disease characteristics associated with delayed diagnosis of infectious pulmonary tuberculosis (TB). METHODOLOGY: A retrospective analysis of 375 adult patients with culture-positive pulmonary TB and cough, treated at the Singapore Tuberculosis Control Unit (TBCU) in 2000, was carried out using data extracted from the TB notification registry and clinical records of the TBCU. Demographic, social, clinical and disease characteristics of patients with reported cough of duration less than, and exceeding, the median duration for the study population were compared. RESULTS: The median duration of cough reported at TB notification was 4 weeks (range, 1-156 weeks). By multivariate analysis, patients with cough > 4 weeks were more likely to be < 65 years old (adjusted odds ratio (OR), 1.8; 95% CI, 1.1-2.9; P = 0.02), of Chinese ethnicity (adjusted OR, 2.0; 95% CI, 1.2-3; P = 0.004), more likely to be sputum acid-fast bacilli smear-positive (adjusted OR, 1.7; 95% CI, 1.1-2.7; P = 0.016), and to have weight loss (adjusted OR, 2.6; 95% CI, 1.7-4; P < 0.01). CONCLUSION: Further studies are needed to identify the possible reasons for delayed diagnosis of TB among those < 65 years old, in the Chinese population in Singapore. PMID- 15823186 TI - Importance of atopic cough, cough variant asthma and sinobronchial syndrome as causes of chronic cough in the Hokuriku area of Japan. AB - OBJECTIVE: A prospective multicentre study was conducted to elucidate the causes of chronic cough in Japan. METHODOLOGY: All consecutive and unselected patients complaining of cough lasting 8 weeks or more, who visited our clinics from 1 June to 31 December 2001, were registered. The causes of chronic cough were diagnosed based on the criteria for definite and probable causes of cough as recommended by the Japanese Cough Research Society. RESULTS: Of the 248 patients enrolled, 72 patients (29.0%) were unavailable for follow up before their diagnostic assessment had been finalized. Among the 176 patients who were adequately assessed, a diagnosis was made in 165 patients (93.7%) either as single cause or as one of two causes: atopic cough in 48 (29.1%) and 11 patients (6.7%); cough variant asthma in 46 (27.9%) and nine patients (5.5%); cough predominant asthma in 14 (8.5%) and three patients (1.8%); and sinobronchial syndrome (SBS) in 28 (17.7%) and 14 patients (8.5%), respectively. A diagnosis of gastro-oesophageal reflux-associated cough was made in a total of four patients (2.4%). CONCLUSION: Atopic cough, asthmatic cough consisting of cough variant asthma and cough predominant asthma, and SBS are major causes of chronic cough in Japan. PMID- 15823187 TI - Usefulness of the Japanese Respiratory Society guidelines for community pneumonia: a retrospective analysis of community-acquired pneumonia between 2000 and 2002 in a general hospital. AB - OBJECTIVE: The aim of this study was to investigate the causative organisms of community-acquired pneumonia (CAP) diagnosed between 2000 and 2002 and to evaluate the Japanese Respiratory Society (JRS) guidelines. METHODOLOGY: A total of 124 cases of CAP diagnosed during the study period were analyzed, and the results were compared with those of a previous study by the authors' research group. Determination of the causative organisms of CAP was based on Gram stain, morphology of colonies, quantitative culture of sputum, identification of bacterial isolates, and serological tests. RESULTS: During the study period, the causative organisms were identified in 42 cases (33.8%). Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis were the major causative organisms. Patients were classified into three groups based on the severity of CAP according to the JRS guidelines. The survival rates of patients with moderate and severe CAP were significantly lower than those of the mild group as evaluated by the Kaplan-Meier method (moderate vs mild, 70% vs 100%; severe vs mild, 40% vs 100%; P < 0.001 for both). Seven patients died during the study, and the risk factors were old age, bedridden status with cerebral infarction, and microaspiration, which was associated with recurrent pneumonia within 17 days. CONCLUSION: This study indicates that the JRS guidelines for CAP are useful for treating patients with CAP in Japan. PMID- 15823188 TI - Adequacy of outpatient management of asthma patients admitted to a state hospital in Argentina. AB - OBJECTIVE: The aim of this study was to assess chronic outpatient management of adult patients admitted with asthma. METHODOLOGY: A cross-sectional survey was conducted of 98 consecutive asthma admissions to a specialized pulmonary State Hospital in Buenos Aires, Argentina, over a 12-month period. Patients were surveyed, within 48 h of admission, with a previously validated questionnaire which deals with chronic outpatient management and measures taken by patients or physicians to treat symptoms during asthma exacerbations. RESULTS: FEV1% predicted was 30.2 +/- 10.7. Mean admission rate and emergency department (ED) visits in the previous year were 0.7 +/- 1.2 and 4.6 +/- 5.1, respectively. A total of 96, 65 and 9% of the patients had been treated previously in the ED, admitted to hospital or mechanically ventilated, respectively. Only 62% had been prescribed inhaled corticosteroids (IC) by their physician; 38% had been prescribed nebulized beta agonists (Nbeta2) and 68% a metered dose inhaler (MDIbeta2). Inhaled beta2-agonist usage during acute exacerbations over the 24 h prior to admission was 14.4 +/- 7.4 puffs for MDIbeta2 and 8.6 +/- 5.4 occasions for Nbeta2. Only 11% of the patients were able to perform all the steps of the MDI inhalation technique correctly. An action plan had been provided by their physicians to 43% of patients, while 58% changed their medication on their own. Only three patients had a peak flow meter (PFM) prescribed. ED was used by 26% for their routine care. No health insurance coverage was available to 75.5% of the patients. CONCLUSIONS: Underuse of IC, poor MDI inhalation technique, and low prescription of an action plan was common and a PFM was seldom prescribed. During exacerbations, many patients changed their medication spontaneously and MDIbeta2 underuse was observed. PMID- 15823189 TI - Alpha-antitrypsin genotypes in Korean patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: Alpha1-antitrypsin (AAT) deficiency is a recognized susceptible factor for chronic obstructive pulmonary disease (COPD) in Western countries, but its importance in Korea is unclear. To date, no definitive case of alpha1-antitrypsin deficiency has been reported in Korea. This study aimed to clarify whether alpha1 antitrypsin deficiency exists and to determine the distribution of alpha1 antitrypsin alleles in the Korean population. METHODOLOGY: The serum concentrations of alpha1-antitrypsin were determined and polymorphisms of the alpha1-antitrypsin gene in 114 COPD patients and in 196 healthy controls were examined. Phenotyping by isoelectric focusing and the genotyping of alpha1 antitrypsin gene by polymerase chain reaction and restriction fragment length polymorphism were performed. RESULTS: No alpha1-antitrypsin level abnormality was found in the patients. M1(Val)/M1(Val) was found to be the most frequent genotype in both groups (69.2% and 66.8%, respectively), and M1(Val) the most frequent allele. The distributions of alpha1-antitrypsin alleles were similar in the patient and control groups, and no S or Z allele was found. CONCLUSION: Alpha1 antitrypsin deficiency is unlikely to be an important cause of chronic obstructive pulmonary disease in the Korean population. PMID- 15823190 TI - An interesting cause of recurrent haemoptysis: haemoptysis 7 years after a foreign body penetrated the lung parenchyma and aorta. AB - A 43-year-old man presented with a 12-month history of recurrent haemoptysis. Postero-anterior chest X-ray of a patient with a history of a penetrating thoracic trauma 8 years previously showed a long wedge-shaped opacity just above the left hemidiaphragm, representing the 'tip of the knife' appearance, and penetrating from the lateral chest wall deep to the thoracic aorta. After consultation with the cardiovascular surgeons, it was decided that the patient should have an operation to remove the foreign body penetrating the aorta. During the operation, a piece of glass was located in the posterior segment of the left lower lobe, and it had also penetrated the aorta through to the posterior wall. The glass had a pointed end, was wedge-shaped and measured 8 cm x 3 cm x 0.5 cm. It was removed, and a 5-cm segment of aorta was replaced with dacron graft. Patients with penetrating chest trauma require routine chest X-rays as many will have a haemothorax, pneumothorax or a penetrating foreign body in the chest in the absence of clinical findings. Postero-anterior chest X-rays as well as lateral X-rays must be carefully and systematically examined for foreign bodies. PMID- 15823191 TI - Smoking reduction does work: resulting alterations in the incidence and histological subtypes of lung cancer in New South Wales in the last 20 years. AB - BACKGROUND: There is irrefutable evidence of the association between lung cancer and smoking. The effects of public health campaigns to reduce population-smoking rates on the incidence and the histological distribution of lung cancer were examined. METHODS: The data held in the New South Wales Cancer Registry from 1972 to 2001 was accessed. RESULTS: The data revealed a decreasing incidence of lung cancer amongst males and a rate that continued to increase amongst females. Interestingly, there was also an effect on the histological distribution of lung cancers, with falling rates of small cell lung cancer and squamous cell cancer, both of which have a high association with smoking; and an increasing incidence of adenocarcinomas. CONCLUSIONS: These trends reveal patterns seen worldwide. The increasing proportion of adenocarcinomas in particular may be related to the changing composition of cigarettes and filters. PMID- 15823192 TI - The effect of a post acute respiratory outreach service for patients with chronic obstructive pulmonary disease on hospital readmission rates. AB - OBJECTIVE: The aim of this study was to investigate the effect of a Post Acute Respiratory Outreach Service (PAROS) for patients with chronic obstructive pulmonary disease (COPD), on hospital utilization. METHODS: A retrospective cohort of patients was examined, using a nested comparison, to test the hypothesis that PAROS would affect hospital utilization in the 12 months following the intervention. Patients admitted with COPD and subsequently enrolled in PAROS were compared with age, sex and diagnosis-matched controls admitted with COPD, who did not receive PAROS. RESULTS: Of 216 patients admitted with COPD during the 1-year study period, 28 were referred to PAROS. A total of 25 suitable controls were identified. Three cases that could not be matched were excluded from the analysis. There were no significant differences in lung function or prior hospitalization between the two groups at baseline. A significant increase in hospital bed days, in the 12 months following the index admission, was observed in the PAROS group (P = 0.046). There was no difference in the number of admissions or emergency department presentations between the groups. CONCLUSIONS: The PAROS program did not lead to a reduction in hospital utilization. This study supports previous findings that respiratory outreach services delivered immediately post discharge increase hospital utilization. PMID- 15823193 TI - Prevalence of asthma, eczema and allergic rhinitis: two surveys, 6 years apart, in Kota Bharu, Malaysia. AB - OBJECTIVE: This study was undertaken to determine the prevalence of asthma, eczema, and allergic rhinitis in school children in Kota Bharu, Malaysia, and in so doing to determine the differences in symptom prevalence rates of asthma, and atopic diseases in Kota Bharu school children between 1995 and 2001. METHODOLOGY: In two studies (1995 and 2001), year one primary school (PS) pupils (6-7 years old) and secondary school (SS) year two pupils (13-14 years old) were randomly selected from the district of Kota Bharu, Kelantan, Malaysia. In 1995, 3939 PS children and 3116 SS children participated, and in 2001 3157 PS children and 3004 SS children participated. The Phase I International Study of Asthma and Allergies in Childhood prevalence written questionnaire and video questionnaire (only shown to SS children) were used in both studies. RESULTS: The written questionnaire showed no significant changes in the prevalence (1995, 2001) of ever wheeze (8.3%, 6.9%P = 0.06), current wheeze (5.4%, 4.3%P = 0.08), exercise-induced wheeze (EIW; 3.9%, 3.7%P = 0.63), and rhinoconjunctivitis (4.6%, 5%P = 0.42) among PS children. The prevalence of flexural itchy rash increased from 14% to 17.6% (P = 0.004) and night cough decreased from 20.4% to 17.5% (P = 0.005). There were also no significant changes in these symptoms among SS children (1995, 2001): ever wheeze (10.7%, 12%P = 0.37), current wheeze (6.8%, 5.7%P = 0.20), EIW (9.9%, 11.6%P = 0.28), night cough (21.6%, 24%P = 0.39), rhinoconjunctivitis (11%, 15%P = 0.11), and flexural itchy rash (12%, 13%P = 0.11). The video questionnaire showed no significant changes in the prevalence of symptoms in the previous 12 months (1995 vs 2001) for wheeze at rest (3.8%, 2.8%P = 0.12), EIW (6.9%, 8.8%P = 0.32), waking with wheeze (1.7%, 1.7%P = 1.0), and severe wheeze (2.1%, 3%P = 0.12). Night cough in the previous 12 months increased significantly from 5.1% to 8.3% (P = 0.007). CONCLUSION: Although asthma and atopic disorders are common in this country, the results revealed no major changes in the prevalence rates of these diseases over a period of 6 years. PMID- 15823194 TI - A case report of congenital isolated absence of the right pulmonary artery: bronchofibrescopic findings and chest radiological tracings over 9 years. AB - A 27-year-old man was admitted to hospital for investigation of haemoptysis. He was a serving member of the Japan Self-Defense Forces. A CXR showed absence of the right hilum, and the right hemithorax was smaller than the contralateral hemithorax. Pulmonary arteriography demonstrated complete absence of the right pulmonary artery. Right heart catheterisation did not show any concomitant cardiovascular malformations, and the patient was diagnosed as having isolated absence of the right pulmonary artery. Bronchial arteriograms demonstrated enlargement and proliferation of the small branches of the right bronchial artery. Bronchofibrescopic examination showed obvious enlargement of the bronchial mucosal vessels with submucosal eruptions and swelling in the right bronchial tree, but not on the left side. Re-examination of the patient's CXR from the previous 9 years revealed a chronological decrease of right lung volume and an increase of the cardiothoracic ratio. Isolated absence of the right pulmonary artery is generally considered to have a good prognosis, but close observation is necessary to monitor the pulmonary haemodynamics. PMID- 15823195 TI - Clinical presentation of metabolic alkalosis in an adult patient with cystic fibrosis. AB - In subtropical and tropical climates, dehydration is common in cystic fibrosis patients with respiratory exacerbations. This may lead to a clinical presentation of metabolic alkalosis with associated hyponatraemia and hypochloraemia. An adult cystic fibrosis patient who presented with a severe respiratory exacerbation accompanied by metabolic alkalosis is presented and the effects of volume correction are reported. PMID- 15823196 TI - Tuberculous peritonitis developing during chemotherapy for pulmonary and intestinal tuberculosis: a case report. AB - This report is of a case of tuberculous peritonitis that developed during antituberculous chemotherapy. A 54-year-old man had been diagnosed as all-drug susceptible pulmonary and intestinal tuberculosis, and treatment with isoniazid, ethambutol and rifampicin had been initiated. About 5 months later, while still undergoing therapy, a large amount of ascites developed. A diagnostic laparoscopy was performed but due to the adhesion between the greater omentum and the parietal peritoneum, intestinal perforation occurred. An emergency operation was performed and the diagnosis of tuberculous peritonitis was confirmed. There are few reports of abdominal tuberculosis developing during antituberculous chemotherapy. In this case a paradoxical response may have been involved in the pathogenesis. PMID- 15823197 TI - Pulmonary anisakiasis presenting as eosinophilic pleural effusion. AB - A 63-year-old man developed a pleural effusion with marked eosinophilia, which was more prominent in the pleural fluid than in the peripheral blood. The pleural effusion spontaneously disappeared 7 days after admission. A multiple dot enzyme linked immunosorbent assay for anisakiasis was strongly positive for both the serum and pleural fluid. The serum IgG titre for Anisakis simplex gradually decreased over 7 months. It is suspected that Anisakis larvae can penetrate the alimentary canal, and then migrate into the pleural cavity through the diaphragm. Screening with a serological test is useful in the diagnosis of this condition; human pulmonary anisakiasis. PMID- 15823199 TI - Physicochemical characterization of vibriophage N5. AB - Phage N5 is one of the phages of Vibrio cholerae serovar O1 biotype El Tor (Ghosh, A. N., Ansari, M. Q., and Dutta, G. C. Isolation and morphological characterization of El Tor cholera phages. J. Gen. Virol. 70: 2241-2243, 1989). In the present communication the growth curve, molecular weight and confirmation of the genome, partial denaturation map and restriction endonuclease digestion pattern have been determined. Partial denaturation map indicates that the genome has non-permuted / invariant sequence. Presence of cohesive ends has also been documented. PMID- 15823198 TI - Goat uromodulin promoter directs kidney-specific expression of GFP gene in transgenic mice. AB - BACKGROUND: Uromodulin is the most abundant protein found in the urine of mammals. In an effort to utilize the uromodulin promoter in order to target recombinant proteins in the urine of transgenic animals we have cloned a goat uromodulin gene promoter fragment (GUM promoter) and used it to drive expression of GFP in the kidney of transgenic mice. RESULTS: The GUM-GFP cassette was constructed and transgenic mice were generated in order to study the promoter's tissue specificity, the GFP kidney specific expression and its subcellular distribution. Tissues collected from three GUM-GFP transgenic mouse lines, and analyzed for the presence of GFP by Western blotting and fluorescence confirmed that the GUM promoter drove expression of GFP specifically in the kidney. More specifically, by using immuno-histochemistry analysis of kidney sections, we demonstrated that GFP expression was co-localized, with endogenous uromodulin protein, in the epithelial cells of the thick ascending limbs (TAL) of Henle's loop and the early distal convoluted tubule in the kidney. CONCLUSION: The goat uromodulin promoter is capable of driving recombinant protein expression in the kidney of transgenic mice. The goat promoter fragment cloned may be a useful tool in targeting proteins or oncogenes in the kidney of mammals. PMID- 15823200 TI - New Vibrio cholerae O1 biotype ElTor bacteriophages. AB - We report the presence of three new O1 ElTor vibriophages named AS1, AS2 and AS3, isolated from the sewage and pond waters of the outskirts of Kolkata. A few phages, named AS4, with hexagonal heads and abnormally long tails with typical curly projections were also found in the water samples. PMID- 15823201 TI - Localization and gene expression of steroid sulfatase by RT-PCR in cumulus cells and relationship to serum FSH levels observed during in vitro fertilization. AB - BACKGROUND: The purpose of this study was to localize the expression of steroid sulfatase (STS) in cumulus cells and to determine the relationship between STS mRNA expression and the serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol and progesterone. METHODS: The subject group included 49 women (29 to 44 years old) for whom in vitro fertilization treatment was indicated. All subjects gave informed consent. One hundred fourteen samples of cumulus-oocyte complex (COC) were obtained under microscopic observation. Part of the COC was stained by STS antibody. RNA was extracted by phenol-chloroform method and real-time PCR was performed. Serum of each patient was collected and was measured by ELISA. RESULTS: Some of the cumulus samples were stained by STS antibody. The expression of STS mRNA in all samples was confirmed by quantitative RT-PCR. Although there was no significant correlation between the level of STS mRNA and the serum levels of estradiol, progesterone and LH, there was a statistically significant negative correlation between the level of STS mRNA expression and the serum level of FSH (n = 105, p = 0.018, r = -0.22). CONCLUSION: These results have demonstrated for the first time the expression of STS in cumulus cells by immunohistological stainings and real-time RT-PCR. STS expression in cumulus cells may be related to the control of the local steroidal environment in the oocyte. Serum FSH may control STS mRNA expression from the results of RT-PCR, although the correlation was low. PMID- 15823202 TI - Differential selection and mutation between dsDNA and ssDNA phages shape the evolution of their genomic AT percentage. AB - BACKGROUND: Bacterial genomes differ dramatically in AT%. We have developed a model to show that the genomic AT% in rapidly replicating bacterial species can be used as an index of the availability of nucleotides A and T for DNA replication in cellular medium. This index is then used to (1) study the evolution and adaptation of the bacteriophage genomic AT% in response to the differential nucleotide availability of the host and (2) test the prediction that double-stranded DNA (dsDNA) phage should exhibit better adaptation than single stranded DNA (ssDNA) phage because the rate of spontaneous deamination, which leads to C-->T or C-->U mutations depending on whether C is methylated or not, is about 100-fold greater in ssDNA than in dsDNA. RESULTS: We retrieved 79 dsDNA phage and 27 ssDNA phage genomes together with their host genomic sequences. The dsDNA phages have their genomic AT% better adapted to the host genomic AT% than ssDNA phage. The poorer adaptation of the ssDNA phage can be partially accounted for by the C-->T(U) mutations mediated by the spontaneous deamination. For ssDNA phage, the genomic A% is more strongly correlated with their host genomic AT% than the genomic T%. CONCLUSION: A significant fraction of variation in the genomic AT% in the dsDNA phage, and that in the genomic A% and T% of the ssDNA phage, can be explained by the difference in selection and mutation between them. PMID- 15823203 TI - Analysis of sequence variability in the CART gene in relation to obesity in a Caucasian population. AB - BACKGROUND: Cocaine and amphetamine regulated transcript (CART) is an anorectic neuropeptide located principally in hypothalamus. CART has been shown to be involved in control of feeding behavior, but a direct relationship with obesity has not been established. The aim of this study was to evaluate the effect of polymorphisms within the CART gene with regards to a possible association with obesity in a Caucasian population. RESULTS: Screening of the entire gene as well as a 3.7 kb region of 5' upstream sequence revealed 31 SNPs and 3 rare variants; 14 of which were subsequently genotyped in 292 French morbidly obese subjects and 368 controls. Haplotype analysis suggested an association with obesity which was found to be mainly due to SNP-3608T>C (rs7379701) (p = 0.009). Genotyping additional cases and controls also of European Caucasian origin supported further this possible association between the CART SNP -3608T>C T allele and obesity (global p-value = 0.0005). Functional studies also suggested that the SNP 3608T>C could modulate nuclear protein binding. CONCLUSION: CART SNP -3608T>C may possibly contribute to the genetic risk for obesity in the Caucasian population. However confirmation of the importance of the role of the CART gene in energy homeostasis and obesity will require investigation and replication in further populations. PMID- 15823204 TI - The effect of executive walk rounds on nurse safety climate attitudes: a randomized trial of clinical units[ISRCTN85147255] [corrected]. AB - BACKGROUND: Executive walk rounds (EWRs) are a widely used but unstudied activity designed to improve safety culture in hospitals. Therefore, we measured the impact of EWRs on one important part of safety culture -- provider attitudes about the safety climate in the institution. METHODS: Randomized study of EWRs for 23 clinical units in a tertiary care teaching hospital. All providers except physicians participated. EWRs were conducted at each unit by one of six hospital executives once every four weeks for three visits. Providers were asked about their concerns regarding patient safety and what could be done to improve patient safety. Suggestions were tabulated and when possible, changes were made. Provider attitudes about safety climate measured by the Safety Climate Survey before and after EWRs. We report mean scores, percent positive scores (percentage of providers who responded four or higher on a five point scale (agree slightly or agree strongly), and the odds of EWR participants agreeing with individual survey items when compared to non-participants. RESULTS: Before EWRs the mean safety climate scores for nurses were similar in the control units and EWR units (78.97 and 76.78, P = 0.458) as were percent positive scores (64.6% positive and 61.1% positive). After EWRs the mean safety climate scores were not significantly different for all providers nor for nurses in the control units and EWR units (77.93 and 78.33, P = 0.854) and (56.5% positive and 62.7% positive). However, when analyzed by exposure to EWRs, nurses in the control group who did not participate in EWRs (n = 198) had lower safety climate scores than nurses in the intervention group who did participate in an EWR session (n = 85) (74.88 versus 81.01, P = 0.02; 52.5% positive versus 72.9% positive). Compared to nurses who did not participate, nurses in the experimental group who reported participating in EWRs also responded more favorably to a majority of items on the survey. CONCLUSION: EWRs have a positive effect on the safety climate attitudes of nurses who participate in the walk rounds sessions. EWRs are a promising tool to improve safety climate and the broader construct of safety culture. PMID- 15823205 TI - Host-derived pathogenicity islands in poxviruses. AB - BACKGROUND: Poxviruses are important both as pathogens and as vaccine vectors. Poxvirus genomes (150-350 kb) consist of a single linear dsDNA molecule; the two polynucleotide strands are joined by short hairpin loops. The genomes encode highly conserved proteins required for DNA replication and mRNA transcription as well as a variable set of virulence factors; transcription takes place within the cytoplasm of the host cell. We are interested in evolution of poxvirus genomes and especially how these viruses acquire host-derived genes that are believed to function as virulence factors. RESULTS: Using a variety of bioinformatics tools, we have identified regions in poxvirus genomes that have unusual nucleotide composition (higher or lower than average A+T content) compared to the genome as a whole; such regions may be several kilobases in length and contain a number of genes. Regions with unusual nucleotide composition may represent genes that have been recently acquired from the host genome. The study of these genomic regions with unusual nucleotide content will help elucidate evolutionary processes in poxviruses. CONCLUSION: We have found that dotplots of complete poxvirus genomes can be used to locate regions on the genome that differ significantly in A+T content to the genome as a whole. The genes in these regions may have been acquired relatively recently from the host genome or from another AT-rich poxvirus. PMID- 15823206 TI - Sodium pertechnetate (Na99mTcO4) biodistribution in mice exposed to cigarette smoke. AB - BACKGROUND: The biological effects of cigarette smoke are not fully known. To improve our understanding of the action of various chemical agents, we investigated the biodistribution of sodium pertechnetate (Na99mTcO4) in mice exposed to cigarette smoke. METHODS: Fifteen BALB/c male mice were exposed to the smoke of nine whole commercial cigarettes per day, 3 times/day, for up to 10 days to whole body exposure in a chamber. A control group of 5 BALB/c male mice was sham-smoked. One day later, the exposed and control groups of mice received (7.4 MBq/0.3 ml) of Na99mTcO4 before being killed at 30 min. Bones, brain, heart, intestine, kidney, liver, lungs, muscle, pancreas, spleen, stomach, testis and thyroid were weighed and these organs and blood radioactivity recorded with a gamma counter. The percentage per gram of tissue of injected dose (%ID/g) was determined for each organ. RESULTS: Cigarette smoke significantly decreased (p < 0.05) the %ID/g in red blood cells, bone, kidney, lung, spleen, stomach, testis and thyroid of the exposed mice. CONCLUSION: The toxic effects of cigarette smoke reduced the Na99mTcO4 biodistribution. PMID- 15823207 TI - Development and evaluation of a hand held computer based on-call pack for health protection out of hours duty: a pilot study. AB - BACKGROUND: The on call service for health protection in most parts of the UK is provided by general public health consultants, registrars and nurses as the first tier of response backed up by medical consultants in health protection. The first tier responder usually carries a large bag of papers containing both local and national guidance on the management of common cases/incidents. An electronic on call pack may provide a suitable practical alternative to large paper based systems and help professionals deliver out of hours health protection advice and response to incidents. METHODS: We developed and piloted an electronic on call pack in Hertfordshire for use at the health protection unit level containing key local and national guidelines, contact information and useful references. The on call pack was initially piloted using a laptop and more recently using a personal digital assistant (PDA). The use of the on-call pack was evaluated. RESULTS: Key advantages of the electronic system include reduced size, faster access to information that is clearly indexed and the relative ease of updating information. As part of the pilot, the electronic on call pack was presented to a local and regional training meeting with good response from participants using qualitative and quantitative methods. CONCLUSION: It is anticipated that with suitable evaluation this system can be adapted and utilised by other health protection practitioners. This system provides a fast, reliable and easily maintained source of information for the public health on-call team. PMID- 15823208 TI - Airway allergen exposure stimulates bone marrow eosinophilia partly via IL-9. AB - BACKGROUND: Interleukin (IL)-9 is a Th2-derived cytokine with pleiotropic biological effects, which recently has been proposed as a candidate gene for asthma and allergy. We aimed to evaluate the therapeutic effect of a neutralizing anti-IL-9 antibody in a mouse model of airway eosinophilic inflammation and compared any such effect with anti-IL-5 treatment. METHODS: OVA-sensitized Balb/c mice were intraperitoneally pretreated with a single dose (100 microg) of an anti mouse IL-9 monoclonal antibody (clone D9302C12) or its vehicle. A third group was given 50 microg of a monoclonal anti-mouse IL-5 antibody (TRFK-5) or its vehicle. Animals were subsequently exposed to OVA on five days via airways. Newly produced eosinophils were labelled using 5-bromo-2'-deoxyuridine (BrdU). BrdU+ eosinophils and CD34+ cell numbers were examined by immunocytochemistry. After culture and stimulation with OVA or PMA+IC, intracellular staining of IL-9 in bone marrow cells from OVA-exposed animals was measured by Flow Cytometry. The Mann-Whitney U test was used to determine significant differences between groups. RESULTS: Anti IL-9 significantly reduced bone marrow eosinophilia, primarily by decrease of newly produced (BrdU+) and mature eosinophils. Anti-IL-9 treatment also reduced blood neutrophil counts, but did not affect BAL neutrophils. Anti-IL-5 was able to reduce eosinophil numbers in all tissue compartments, as well as BrdU+ eosinophils and CD34+ progenitor cells, and in all instances to a greater extent than anti-IL-9. Also, FACS analysis showed that IL-9 is over-expressed in bone marrow CD4+ cells after allergen exposure. CONCLUSIONS: Our data shows that a single dose of a neutralizing IL-9 antibody is not sufficient to reduce allergen induced influx of newly produced cells from bone marrow to airways. However, in response to allergen, bone marrow cells over-express IL-9. This data suggest that IL-9 may participate in the regulation of granulocytopoiesis in allergic inflammation. PMID- 15823209 TI - Quantitative assessment of the effect of uracil-DNA glycosylase on amplicon DNA degradation and RNA amplification in reverse transcription-PCR. AB - Although PCR and RT-PCR provided a valuable approach for detection of pathogens, the high level of sensitivity of these assays also makes them prone to false positive results. In addition to cross-contamination with true positive samples, false positive results are also possible due to "carry-over" contamination of samples with amplicon DNA generated by previous reactions. To reduce this source of false positives, amplicon generated by reactions in which dUTP was substituted for dTTP can be degraded by uracil DNA glycosylase (UNG). UNG does not degrade RNA but will cleave contaminating uracil-containing DNA while leaving thymine containing DNA intact. The availability of heat-labile UNG makes use of this approach feasible for RT-PCR. In this study, real-time RT-PCR was used to quantify UNG degradation of amplicon DNA and the effect of UNG on RNA detection. Using the manufacturers' recommended conditions, complete degradation of DNA was not observed for samples containing 250 copies of amplicon DNA. Doubling the UNG concentration resulted in degradation of the two lowest concentrations of DNA tested, but also resulted in an increase of 1.94 cycles in the CT for RNA detection. To improve DNA degradation while minimizing the effect on RNA detection, a series of time, temperature and enzyme concentrations were evaluated. Optimal conditions were found to be 0.25 U UNG per 25 microl reaction with a 20 min, 30 degrees C incubation prior to RT-PCR. Under these conditions, high concentrations of amplicon DNA could be degraded while the CT for RNA detection was increased by 1.2 cycles. PMID- 15823210 TI - Intrapulmonal dislocation of a totally implantable venous access device. AB - BACKGROUND: Totally implantable venous access devices are widely used for infusion of chemotherapy or parenteral nutrition. Device associated complications include technical operative problems, infections, paravasal infusions and catheter or punction chamber dislocation. CASE PRESENTATION: We present the case of a 49-year-old patient with the rare complication of a intrapulmonal catheter dislocation of a totally implantable venous access system. Treosulfane for chemotherapy of metastatic breast cancer was infused via the catheter causing instant coughing and dyspnoea which lead to the diagnosis of catheter dislocation. The intrapulmonal part of the catheter was removed under thoracoscopic control without further complications. CONCLUSION: Intrapulmonal catheter dislocation is a rare complication of a totally implantable venous access device which can not be avoided by any prophylactic measures. Therefore, the infusion system should be tested before each use and each new symptom, even when not obviously related to the catheter should be carefully documented and evaluated by expert physicians to avoid severe catheter-associated complications. PMID- 15823211 TI - A cluster randomized controlled trial of a behavioral intervention to facilitate the development and implementation of clinical practice guidelines in Latin American maternity hospitals: the Guidelines Trial: Study protocol [ISRCTN82417627]. AB - BACKGROUND: A significant proportion of the health care administered to women in Latin American maternity hospitals during labor and delivery has been demonstrated to be ineffective or harmful, whereas effective interventions remain underutilized. The routine use of episiotomies and the failure to use active management of the third stage of labor are good examples. METHODS/DESIGN: The aim of this trial is to evaluate the effect of a multifaceted behavioral intervention on the use of two evidence-based birth practices, the selective use of episiotomies and active management of the third stage of labor (injection of 10 International Units of oxytocin). The intervention is based on behavioral and organizational change theories and was based on formative research. Twenty-four hospitals in three urban districts of Argentina and Uruguay will be randomized. Opinion leaders in the 12 intervention hospitals will be identified and trained to develop and implement evidence-based guidelines. They will then disseminate the guidelines using a multifaceted approach including academic detailing, reminders, and feedback on utilization rates. The 12 hospitals in the control group will continue with their standard in-service training activities. The main outcomes to be assessed are the rates of episiotomy and oxytocin use during the third stage of labor. Secondary outcomes will be perineal sutures, postpartum hemorrhages, and birth attendants' opinions. PMID- 15823212 TI - High versus standard doses interferon-alpha in the treatment of naive chronic hepatitis C patients in Taiwan: a 10-year cohort study. AB - BACKGROUND: Interferon-alpha monotherapy is effective in less than one-third patients with chronic hepatitis C. The dose-effect, tolerability and durability of interferon-alpha treatment and its long-term effect on the prevention of cirrhosis and hepatocellular carcinoma in naive Taiwanese patients with chronic hepatitis C have not been well investigated. We conducted the present cohort study treated with high and standard interferon-alpha to illustrate the issues. METHODS: We performed a long-term virologic and histological follow-up of 214 chronic hepatitis C patients treated with interferon-alpha, 3 million units (3 MU, n = 80) or 6-MU (n = 134) thrice weekly for 24 weeks, in Taiwan between 1992 and 2001. RESULTS: There was no difference in the incidence of discontinuation between 3-MU and 6-MU groups (4/80, 5.0% versus 10/134. 7.5%). The 6-MU group had similar incidence of adverse events with the 3-MU group, except that 6-MU group had significantly higher incidence of psychological manifestations, mainly presented as irritability. The rates of sustained virological response (SVR) were significantly higher in 6-MU regimen (37.1%) than in 3-MU regimen (23.7%, p < 0.05) in per protocol analysis. Based on multivariate analysis, baseline viral load was strongly associated with SVR, followed by hepatitis C virus genotype, interferon-alpha regimen, and liver fibrosis. A histological improvement in necroinflammatory activity, but not in fibrosis was observed in the follow-up biopsy performed 0.5-5.5 years (mean: 1.9 years, n = 51) after end-of-treatment. Among patients without SVR, there was more activity improvement in 6-MU group. The durability of SVR was 100% (18/18) and 97.8% (45/46) for 3-MU and 6-MU group, respectively, in a mean follow-up period of 6.81 years (5.25-9.18 years). For 163 baseline non-cirrhotic patients, 9 of 84 (10.7%) non-responders and 3 of 79 (3.8%) sustained responders progressed to cirrhosis during a mean follow-up period of 5.52 and 5.74 years, respectively (p = 0.067, Kaplan-Meier survival analysis, log-rank test). For all 200 patients, hepatocellular carcinoma was detected in 12 of 113 (10.6%) non-responders and one of 87 (1.1%) sustained responders during a mean follow-up period of 5.67 and 5.73 years, respectively (p < 0.01, Kaplan-Meier survival analysis, log-rank test). CONCLUSION: We confirm the dose effect of interferon-alpha in chronic hepatitis C. Six-MU regimen had better efficacy than 3-MU regimen in virologic and histological responses. Both regimens had good tolerability and durability in Taiwan. Sustained response could reduce the incidence of cirrhotic change and hepatocarcinogenesis. PMID- 15823214 TI - Statistical analysis of data in support of the Barker hypothesis, advantages of using random effects regression model in hierarchical data. AB - In a recent note by Walters and Edwards (2004), the authors argued that summary statistics should be used in analysing hierarchical data from our earlier analysis of a rat model of developmental programming and the Barker hypothesis (Kwong et al., 2000, 2004). We reiterate here why such a view is inappropriate. Hierarchical data merits multilevel analysis using a 'random effects' model to enable estimation of variances at different levels and easy assessment of other parameters in a complex data structure. PMID- 15823215 TI - HLA-matched embryos selected for siblings requiring haematopoietic stem cell transplantation: a psychological perspective. AB - Allogeneic haematopoietic stem cell transplantation (HSCT) is a treatment for a number of acquired and congenital diseases. An important factor in the outcome of the treatment is the degree of human leukocyte antigen (HLA) compatibility between patient and donor. HLA identical siblings therefore provide the best chance for the recipient. Preimplantation genetic diagnosis (PGD) can be used to select HLA identical embryos if there is no HLA compatible sibling in the family. The Centre for Reproductive Medicine considered it morally justified to give medical assistance to couples in need of an HLA matched sibling. Two considerations played an important role in this respect: (i) the use as a donor should not be the only parents' motive for having the child and (ii) IVF and HLA typing on the embryos would be less of a burden for the parents than other alternatives. Since the first request in 2000, 12 couples have been referred for psychological counselling. The motivation of four couples will be discussed in depth. The validity of the arguments will be checked against the experience of the actual cases. The consequences of the treatment on the welfare of the future donor child will be discussed. PMID- 15823217 TI - Spare embryos: 3000 reasons to rethink the significance of genetic relatedness. AB - IVF laws in Victoria, Australia provide that human embryos must not remain in storage in excess of 5 years. Studies reviewing the fate of surplus human embryos reveal that close to 90% of couples choose to discard their excess embryos and that hundreds of embryos are disposed of annually. It has been argued that human embryos are a valuable resource and that there is a need to consider educational programmes to encourage couples to donate spare embryos to other infertile couples, rather than discard them. Surveys show that one reason that so few embryos are donated is that couples attach great significance to genetic parenthood. Advances in reproductive technology may necessitate a review of biological definitions of family and the importance of genetic relatedness. It can be argued that it is unreasonable to conclude that genetic ties are so significant that embryos should be discarded rather than donated and raised by non-genetically related parents. It is suggested that education programmes should encourage reflection on people's beliefs about the importance of genetic relatedness with regard to what makes a family. Open embryo donation or directed embryo donation programmes might cause couples to change their minds, or alleviate their anxiety about donating embryos to others. PMID- 15823219 TI - Continued improvements in the quality and consistency of follitropin alfa, recombinant human FSH. AB - The use of gonadotrophins for the treatment of infertility began in the 1930s following early work on the pituitary-ovarian axis and the discovery of FSH and LH. The technological development of pharmaceutical gonadotrophins over the last 40 years has shown improvements in specific activity, purity, degradation and impurities. Throughout these pharmaceutical developments the gonadotrophin content of both urinary and recombinant preparations has been assessed using an animal in-vivo bioassay. This paper reflects upon the manufacturing history of recombinant human FSH (r-hFSH) and follitropin alfa filled by mass (FbM), and evaluates the impact of introducing a pharmaceutical product that is formulated and assayed by a physicochemical method for r-hFSH protein content. It also compares the analytical consistency of follitropin alfa FbM with another commercially available r-hFSH, follitropin beta. PMID- 15823220 TI - Case report: delirium associated with ovarian hyperstimulation syndrome. AB - Ovarian hyperstimulation syndrome (OHSS) is one of the most important complications of assisted reproductive technologies. Mild OHSS is characterized by ovarian enlargement and abdominal discomfort. In severe cases anasarca, hepatic dysfunction, reduced blood volume, electrolyte imbalance, organ failure and thromboembolic phenomena may be observed. Delirium is a syndrome, not a disease, and has many causes, all of which result in a similar pattern of signs and symptoms relating to a patient's level of consciousness and cognitive impairment. Delirium remains an under-recognized and under-diagnosed clinical disorder. The case is presented of a 30-year-old woman with OHSS and delirium. She underwent intracytoplasmic sperm injection (ICSI) for severe male factor infertility. Five days after oocyte retrieval, ascite formation was observed in ultrasonographic evaluation, and embryo transfer was cancelled. Twelve days after retrieval she came to the emergency clinic with abdominal distension and pain. She was hospitalized and paracentesis was performed every other day three times. She had altered consciousness and psychomotor hypoactivity 1 h after the last paracentesis. Psychiatric consultation revealed that she was in a state of delirium, and haloperidol was administered for treatment. Her symptoms disappeared within a week. Her medication was stopped when symptoms resolved and she was still asymptomatic in psychiatric evaluation 1 week later. PMID- 15823221 TI - Establishment of oocyte population in the fetal ovary: primordial germ cell proliferation and oocyte programmed cell death. AB - Strict control of cell proliferation and cell loss is essential for the coordinated functions of different cell populations in complex multicellular organisms. Oogenesis is characterized by a first phase occurring during embryo fetal life and in common with spermatogenesis, during which mitotic proliferation of the germline stem cells, the primordial germ cells (PGC), prevails over germ cell death. The result is the formation of a relatively high number of germ cells depending on the species, ready to enter sex specific differentiation. In the female, PGC enter into meiosis and become oocytes, thereby ending their stem cell potential. After entering into meiosis in the fetal ovary, oocytes pass through leptotene, zygotene and pachytene stages before arresting in the last stage of meiotic prophase I, the diplotene or dictyate stage at about the time of birth. The most part of oocytes die during the fetal period or shortly after birth. It is widely accepted that in mammals a female is born with a fixed number of oocytes within the ovaries, which over the years progressively decreases without possibility for renewal. Once the oocyte reserve has been exhausted, ovarian senescence, driving what is referred to as the menopause in women, rapidly ensues. The fertile lifespan of a female depends by the size of the oocyte pool at birth and the rapidity of the oocyte pool depletion. Which mechanisms control PGC proliferation? Why do most of the oocytes die during fetal life and what are the mechanisms of such massive degeneration? Is it possible to prolong the lifespan of a female by reducing oocyte lost during the fetal life? This review reports some of the most recent results obtained in an attempt to answer these questions. PMID- 15823222 TI - Meiotic spindle visualization in living human oocytes. AB - A computer-assisted polarization microscopy system (polscope) has made it possible to analyse the meiotic spindle of oocytes subjected to intracytoplasmic sperm injection (ICSI) without affecting their viability. It has been shown that the presence of a detectable birefringent meiotic spindle inside the oocyte cytoplasm of human metaphase II (MII) prepared for ICSI is an indicator of oocyte quality, such as fertilization and developmental ability. Meiotic spindle imaging has also shown that this structure, when detectable, is not always aligned with the first polar body (PB1) in fresh MII oocytes. The relationship between the degree of meiotic spindle deviation from the PB1 location and ICSI outcomes is discussed in this paper. When the meiotic spindle of in-vitro matured oocytes is analysed, it is always found to be aligned with the PB1, suggesting that the misalignment observed in the oocytes matured in vivo results from the PB1 displacement during the manipulations for the cumulus and corona removal. Furthermore, polscope analysis of meiotic spindle changes in living MII oocytes subjected to freezing and thawing procedures has shown that the current techniques of oocyte cryopreservation cause meiotic spindle destruction. The polscope system may assist in the selection of fresh and thawed oocytes for ICSI. PMID- 15823223 TI - Cytogenetic analysis of human somatic cell haploidization. AB - Despite recent interest in the derivation of female and male gametes through somatic cell nuclear transfer, there is still insufficient data on chromosomal analysis of these gametes resulting from haploidization, especially involving a human nuclear donor and recipient oocytes. The objective of this study was to investigate the fidelity of chromosomal separation during haploidization of human cumulus cells by in-vitro matured human enucleated MII oocytes. A total of 129 oocytes were tested 4-7, 8-14, or 15-21 h after nuclear transfer (NT) followed by electro-stimulation, resulting in 71.3% activation efficiency on average. Haploidization was documented by the formation of two separate groups of chromosomes, originating from either polar body/pronucleus (PB/PN), or only 2PN, which were tested by 5-colour FISH, or DNA analysis for copy number of chromosomes 13, 16, 18, 21, 22 and X. Two PN were formed more frequently than PB/PN, irrespective of incubation time. In agreement with recent reports on mouse oocytes, as many as 90.2% of the resulting haploid sets tested showed abnormal chromosome segregation, suggesting unsuitability of the resulting artificial gametes for practical application at the present time. PMID- 15823224 TI - Genetic analyses of reproductive function. PMID- 15823225 TI - Factors controlling blastocyst implantation. AB - Establishment of early pregnancy is promoted by a complex network of signalling molecules that mediate cell-to-cell and cell-to-extracellular matrix communications, in order to manifest controlled invasion of the trophectoderm and successful implantation. During the period known as the 'window of implantation', the endometrium expresses specialized proteins, many of which exhibit potential use as markers of endometrial receptivity. Trophoblast differentiation to the invasive phenotype also depends on the up-regulation of certain peptides and the down-regulation of others. Disruption of each pathway is theoretically possible, and studies in animal models suggest that implantation defects result when the function of these proteins is blocked. Indeed, the implantation process is currently considered the most important limiting factor for the establishment of viable pregnancy. The present overview of the literature reports critical families of molecules located at the embryo-maternal interface and describes the mechanisms of interplay and control. Since these factors are crucial to the process of implantation, targeting them might be a valuable contraceptive tool. Conversely, induction of specific peptides may prove to be beneficial in certain infertility cases. PMID- 15823226 TI - Leptin system in embryo development and implantation: a protein in search of a function. AB - Implantation is a crucial moment in the reproduction process that requires perfect synchronization between the embryo and the maternal endometrium. The embryo must reach the blastocyst stage and the endometrium must be prepared to receive it. An appropriate and specific molecular dialogue must also take place between them. There is ample evidence to show that the leptin system is implicated in this cross-talk. Examples are described. Although there is some controversy surrounding the data, they are supported by the presence of leptin receptor mRNA in mouse and human oocytes and embryos throughout preimplantation development. Otherwise, the leptin mRNA is only detected at the blastocyst stage in both human and mouse. Furthermore, leptin is found at higher concentrations in the conditioned media from competent human blastocysts than in those from arrested embryos, suggesting that this molecule is a marker for blastocyst viability. Given that expression of the leptin receptor increases in the human endometrium during the luteal phase, the secreted leptin could trigger its activation. Finally, leptin and the leptin receptor have been detected in implantation sites. All these findings point to the involvement of the leptin system in the molecular mechanism of the implantation process and embryo development. PMID- 15823228 TI - Implantation failures: success of assisted hatching with quarter-laser zona thinning. AB - Implantation failure after IVF is one of the factors associated with a reduced chance of pregnancy for some patients. Assisted hatching methodologies are designed to facilitate the embryo's escape from the zona pellucida, and this strategy has been suggested as a means of improving pregnancy rates in patients with previous implantation failure. The aim of this prospective and randomized study was to evaluate the efficacy of quarter-laser zona thinning assisted hatching (qLZT-AH) in improving the implantation of embryos in patients with previous implantation failure. A total of 150 patients with a history of previous implantation failure were treated with intracytoplasmic sperm injection, and allocated into two groups: group 1, only one previous implantation failure, and group 2, repeated implantation failures. The patients in each group were randomized at the time of embryo transfer into a control group (no qLZT-AH) or experimental group where qLZT-AH was performed. For patients with repeated implantation failures, the implantation rate in those who received laser-thinned embryos was significantly higher (P = 0.02) than in those whose embryos were not laser-thinned (10.9 and 2.6% respectively). However, this difference was not observed in patients who presented with only one previous implantation failure. The data demonstrate that qLZT-AH is an effective strategy for improving the implantation of embryos in patients with repeated implantation failures. PMID- 15823230 TI - Extrapituitary effects of GnRH antagonists in assisted reproduction: a review. AB - GnRH antagonists are used to prevent premature LH surges in assisted reproduction. The pregnancy rates obtained in several studies have been similar to those obtained in the long protocol with gonadotrophin-releasing hormone (GnRH) agonists. However, lower oestrogen serum concentrations have been observed and fewer oocytes retrieved using GnRH antagonists. Thus, potential effects at extrapituitary GnRH receptors in the ovary and the endometrium have been claimed. The article reviews the currently available data from the literature as well as current investigations with respect to effects of cetrorelix on the ovary and the endometrium. No evidence was found to support the hypothesis of extrapituitary effects of GnRH antagonists such as cetrorelix at doses used in assisted reproduction. PMID- 15823231 TI - Management of poor responders in IVF. AB - Correct controlled ovarian stimulation is of paramount importance in assisted reproductive technologies. Therefore, analysis of the ovarian reserve of the patient is mandatory to tailor the best ovarian stimulation regimen. When the ovarian reserve is reduced, the induction of a multifollicular growth remains a challenge. Several factors could be associated with reduced ovarian response. However, reduced ovarian reserve either in older patients or in young patients represents the most frequent aetiological factor. Whatever is the aetiology, one of the main problems is how to predict a reduced ovarian response, and although several tests have been suggested, no very accurate predictive test is available. A variety of different stimulation protocols have been suggested but the lack of any large-scale, prospective, randomized, controlled trials of the different management strategies and the lack of a uniform definition of the population may result in comparisons of heterogeneous groups of patients, making it difficult to draw any definitive conclusions. Natural cycle IVF may represent an easy and cheap approach in the management of this group of patients. Although no controlled large prospective randomized studies are available to compare the natural IVF procedure with ovarian stimulation IVF in poor responder patients, the efficacy of natural cycle IVF is hampered by high cancellation rates mainly due to untimely LH surge. The use of gonadotrophin-releasing hormone antagonists in the late follicular phase, which reduces the premature LH rise rate, and the improvements in laboratory conditions and fertilization techniques, increase the embryo transfer rates, making this procedure more cost-effective. PMID- 15823233 TI - Intracytoplasmic injection of spermatozoa and spermatogenic cells: its biology and applications in humans and animals. AB - Intracytoplasmic sperm injection (ICSI) has become the method of choice to overcome male infertility when all other forms of assisted fertilization have failed. Animals in which ICSI has produced normal offspring include many species. Success rate with normal spermatozoa is well above 50% in the mouse but ICSI success rates in other animals have been low, ranging from 0.3 to 16.5%. Mouse ICSI revealed that spermatozoa that cannot participate in normal fertilization can produce normal offspring by ICSI, provided their nuclei are genomically intact. Human ICSI using infertile spermatozoa has been highly successful perhaps because of the intrinsic instability of human sperm plasma membrane. The health of children born after ICSI and other assisted fertilization techniques is of major concern. Careful analyses suggest that higher incidences of congenital malformations and/or low birth weights after assisted fertilization are largely attributable to parental genetic background and increased incidence of multiple births, rather than to the techniques of assisted fertilization. Since the physiological and nutritional environments of developing embryos may cause persisting alteration in DNA methylation, extreme caution must be exercised in handling gametes and embryos in vitro. In the mouse, round spermatid injection (ROSI) has been routinely successful but its use in humans is controversial. Whether human ROSI and assisted fertilization involving younger spermatogenic cells are medically safe must be the subject of further investigations. PMID- 15823235 TI - Estimation of genetic parameters for test day records of dairy traits in the first three lactations. AB - Application of test-day models for the genetic evaluation of dairy populations requires the solution of large mixed model equations. The size of the (co)variance matrices required with such models can be reduced through the use of its first eigenvectors. Here, the first two eigenvectors of (co)variance matrices estimated for dairy traits in first lactation were used as covariables to jointly estimate genetic parameters of the first three lactations. These eigenvectors appear to be similar across traits and have a biological interpretation, one being related to the level of production and the other to persistency. Furthermore, they explain more than 95% of the total genetic variation. Variances and heritabilities obtained with this model were consistent with previous studies. High correlations were found among production levels in different lactations. Persistency measures were less correlated. Genetic correlations between second and third lactations were close to one, indicating that these can be considered as the same trait. Genetic correlations within lactation were high except between extreme parts of the lactation. This study shows that the use of eigenvectors can reduce the rank of (co)variance matrices for the test-day model and can provide consistent genetic parameters. PMID- 15823236 TI - A comparison of two methods for prediction of response and rates of inbreeding in selected populations with the results obtained in two selection experiments. AB - Selection programmes are mainly concerned with increasing genetic gain. However, short-term progress should not be obtained at the expense of the within population genetic variability. Different prediction models for the evolution within a small population of the genetic mean of a selected trait, its genetic variance and its inbreeding have been developed but have mainly been validated through Monte Carlo simulation studies. The purpose of this study was to compare theoretical predictions to experimental results. Two deterministic methods were considered, both grounded on a polygenic additive model. Differences between theoretical predictions and experimental results arise from differences between the true and the assumed genetic model, and from mathematical simplifications applied in the prediction methods. Two sets of experimental lines of chickens were used in this study: the Dutch lines undergoing true truncation mass selection, the other lines (French) undergoing mass selection with a restriction on the representation of the different families. This study confirmed, on an experimental basis, that modelling is an efficient approach to make useful predictions of the evolution of selected populations although the basic assumptions considered in the models (polygenic additive model, normality of the distribution, base population at the equilibrium, etc.) are not met in reality. The two deterministic methods compared yielded results that were close to those observed in real data, especially when the selection scheme followed the rules of strict mass selection: for instance, both predictions overestimated the genetic gain in the French experiment, whereas both predictions were close to the observed values in the Dutch experiment. PMID- 15823237 TI - Genomic contributions in livestock gene introgression programmes. AB - The composition of the genome after introgression of a marker gene from a donor to a recipient breed was studied using analytical and simulation methods. Theoretical predictions of proportional genomic contributions, including donor linkage drag, from ancestors used at each generation of crossing after an introgression programme agreed closely with simulated results. The obligate drag, the donor genome surrounding the target locus that cannot be removed by subsequent selection, was also studied. It was shown that the number of backcross generations and the length of the chromosome affected proportional genomic contributions to the carrier chromosomes. Population structure had no significant effect on ancestral contributions and linkage drag but it did have an effect on the obligate drag whereby larger offspring groups resulted in smaller obligate drag. The implications for an introgression programme of the number of backcross generations, the population structure and the carrier chromosome length are discussed. The equations derived describing contributions to the genome from individuals from a given generation provide a framework to predict the genomic composition of a population after the introgression of a favourable donor allele. These ancestral contributions can be assigned a value and therefore allow the prediction of genetic lag. PMID- 15823238 TI - A novel method for the estimation of the relative importance of breeds in order to conserve the total genetic variance. AB - The need for conservation of farm animal genetic resources is widely accepted. A key question is the choice of breeds to be conserved. For this purpose, a core set of breeds was introduced in that the total genetic variance of a hypothetical quantitative trait was maximised (MVT core set). For each breed the relative contribution to the core set was estimated and the breeds were ranked for conservation priority according to their relative contribution. The method was based on average kinships between and within breeds and these can be estimated using genetic marker data. The method was compared to a recently published core set method that maximises the variance of a hypothetical population that could be obtained by interbreeding the conserved breeds (MVO core set). The results show that the MVT (MVO) core set favours breeds with a high (low) within breed kinship that are not related to other breeds. Following this, the MVT core set method suggests conserving breeds that show a large difference in the respective population mean of a hypothetical quantitative trait. This maximises the speed of achieving selection response for this hypothetical selection direction. Additionally, bootstrap based methods for the estimation of the breed's contribution to the core sets were introduced, substantially improving the accuracy of the contribution estimates. PMID- 15823239 TI - Identification and characterization of single nucleotide polymorphisms in 12 chicken growth-correlated genes by denaturing high performance liquid chromatography. AB - The genes that are part of the somatotropic axis play a crucial role in the regulation of growth and development of chickens. The identification of genetic polymorphisms in these genes will enable the scientist to evaluate the biological relevance of such polymorphisms and to gain a better understanding of quantitative traits like growth. In the present study, 75 pairs of primers were designed and four chicken breeds, significantly differing in growth and reproduction characteristics, were used to identify single nucleotide polymorphisms (SNP) using the denaturing high performance liquid chromatography (DHPLC) technology. A total of 283 SNP were discovered in 31 897 base pairs (bp) from 12 genes of the growth hormone (GH), growth hormone receptor (GHR), ghrelin, growth hormone secretagogue receptor (GHSR), insulin-like growth factor I and II (IGF-I and -II), insulin-like growth factor binding protein 2 (IGFBP-2), insulin, leptin receptor (LEPR), pituitary-specific transcription factor-1 (PIT-1), somatostatin (SS), thyroid-stimulating hormone beta subunit (TSH-beta). The observed average distances in bp between the SNP in the 5'UTR, coding regions (non- and synonymous), introns and 3'UTR were 172, 151 (473 and 222), 89 and 141 respectively. Fifteen non-synonymous SNP altered the translated precursors or mature proteins of GH, GHR, ghrelin, IGFBP-2, PIT-1 and SS. Fifteen indels of no less than 2 bps and 2 poly (A) polymorphisms were also observed in 9 genes. Fifty nine PCR-RFLP markers were found in 11 genes. The SNP discovered in this study provided suitable markers for association studies of candidate genes for growth related traits in chickens. PMID- 15823240 TI - Strategic and automatic components in the processing of linguistic spatial relations. AB - The objective of the present study was to determine the extent to which strategies influence the representational format of a linguistic spatial relation. The propositional model assumes that a sentence describing a spatial relation is always represented as a set of propositions, whereas the strategic model claims that a spatial sentence can be represented either as a set of propositions or as a mental image, depending on the strategy (verbal or visual spatial) an individual follows. Participants read a sentence (spatial or non spatial) followed by a picture or sentence, which did or did not exemplify the information of the first sentence. In order to examine the involvement of strategic and automatic components the probability (20% or 80%) of the nature (sentence or picture) of the second stimulus was varied. Participants had slower verification RTs for unexpected stimuli than for expected stimuli, but this cost was significantly larger for an unexpected picture than an unexpected sentence. Furthermore, this asymmetric cost for the unexpected visual-spatial stimulus only occurred with spatial sentences and not with non-spatial sentences. Surprisingly, these data do not support a strictly propositional or a strategic model. Instead, we propose a third option: a dual representational model, in which people automatically represent the spatial sentence propositionally. In addition, depending on the context, a pictorial strategy is employed, which results in a supplementary visual-spatial representation. PMID- 15823241 TI - The enhanced Simon effect for older adults is reduced when the irrelevant location information is conveyed by an accessory stimulus. AB - The Simon effect, better performance when irrelevant stimulus location corresponds with the response location than when it does not, typically is larger for older than younger adults. However, Simon and Pouraghabagher [Simon, R. J., & Pouraghabagher, A. R. (1978). The effect of aging on the stages of processing in a choice reaction time task. Journal of Gerontology, 33, 553-561] found no age difference using an accessory-stimulus Simon task in which the relevant dimension was the color of a visual stimulus and the irrelevant dimension the location of a tone. Experiment 1 confirmed that older adults show a larger Simon effect than younger adults for the visual Simon task and that this age-related deficit is reduced or eliminated for the auditory-accessory task. Experiment 2 provided evidence suggesting that a small part of the age-related deficit in the visual Simon task is due to having to code the location of the relevant stimulus, but Experiment 3 showed that the majority of the deficit is due to the relevant and irrelevant information being conveyed by the same stimulus. Reaction-time distribution analyses show similar functions for younger and older adults, suggesting that the time course of activation is similar for both age groups. PMID- 15823242 TI - Simple arithmetic processing: the question of automaticity. AB - In adult simple arithmetic performance, it is commonly held that retrieval of solutions occurs automatically from a network of stored facts in memory. However, such an account of performance necessarily predicts a uniform reaction time for solution retrieval and is therefore not consistent with the robust finding that reaction time increases with problem size and difficulty. Additionally, past research into arithmetic performance has relied on tasks that may have actually induced and measured attentional processing, thereby possibly confounding previous results and conclusions pertaining to automaticity. The present study therefore, attempted to more reliably assess the influence of automatic processing in arithmetic performance by utilizing a variant of the well established semantic word-priming procedure with a target-naming task. The overall results revealed significant facilitation in naming times at SOAs of 240 and 1000 ms for congruent targets i.e., targets that represented the correct solutions to problems presented as primes (e.g., 6+8 and 14). Significant inhibition in comparison to a neutral condition (0+0 and 17) was also observed at 120 and 240 ms SOAs in naming incongruent targets (e.g., 6+8 and 17). Furthermore, response times were found to vary as a function of both arithmetic fluency and problem size. Differences in performance to addition and multiplication operations and implications for cognitive research and education are considered. PMID- 15823243 TI - The influence of writing practice on letter recognition in preschool children: a comparison between handwriting and typing. AB - A large body of data supports the view that movement plays a crucial role in letter representation and suggests that handwriting contributes to the visual recognition of letters. If so, changing the motor conditions while children are learning to write by using a method based on typing instead of handwriting should affect their subsequent letter recognition performances. In order to test this hypothesis, we trained two groups of 38 children (aged 3-5 years) to copy letters of the alphabet either by hand or by typing them. After three weeks of learning, we ran two recognition tests, one week apart, to compare the letter recognition performances of the two groups. The results showed that in the older children, the handwriting training gave rise to a better letter recognition than the typing training. PMID- 15823245 TI - Encoding specificity manipulations do affect retrieval from memory. AB - In a recent article, P.A. Higham (2002) [Strong cues are not necessarily weak: Thomson and Tulving (1970) and the encoding specificity principle revisited. Memory &Cognition, 30, 67-80] proposed a new way to analyze cued recall performance in terms of three separable aspects of memory (retrieval, monitoring, and report bias) by comparing performance under both free-report and forced report instructions. He used this method to derive estimates of these aspects of memory in an encoding specificity experiment similar to that reported by D.M. Thomson and E. Tulving (1970) [Associative encoding and retrieval: weak and strong cues. Journal of Experimental Psychology, 86, 255-262]. Under forced report instructions, the encoding specificity manipulation did not affect performance. Higham concluded that the manipulation affected monitoring and report bias, but not retrieval. I argue that this interpretation of the results is problematic because the Thomson and Tulving paradigm is confounded, and show in three experiments using a more appropriate design that encoding specificity manipulations do affect performance in forced-report cued recall. Because in Higham's framework forced-report performance provides a measure of retrieval that is uncontaminated by monitoring and report bias it is concluded that encoding specificity manipulations do affect retrieval from memory. PMID- 15823244 TI - Initial storage of unfamiliar objects: examining memory stores with signal detection analyses. AB - The memory source(s) underlying performance of the object decision task were examined. Application of signal detection theory with independent indices of sensitivity and bias indicated that greater object decision sensitivity (possible vs. impossible object discrimination) is observed for primed than for unprimed objects and a greater bias to respond "possible" is observed for primed than for unprimed objects. To test whether a single memory store underlies the two effects, we assessed whether the two effects would be influenced in similar ways by a particular experimental manipulation. Sensitivity did not differ between processing of 180 degrees -rotated primed objects vs. un-rotated primed objects, but bias to respond "possible" differed between those conditions in predictable ways depending on the cerebral hemisphere of direct test presentations. Results support a conciliatory resolution to the debate concerning memory in this task and help to clarify properties of memory stores underlying the initial storage of unfamiliar objects. PMID- 15823246 TI - Simvastatin reduced ischemic brain injury and perfusion deficits in an embolic model of stroke. AB - Simvastatin is cholesterol lowering agent and also a modulator of cytokine in the nervous system. The functional significance and neuroprotectiove mechanism of simvastatins in ischemic brain injury is controversial. The purpose of study is to evaluate the effect of simvastatin on ischemic brain injury and to investigate the perfusion capability of brain microvessels in the ischemic injury. This study included two series of experiments. In the first series, we studied if simvastatin is neuroprotective in an embolic model of stroke. The treatments began 2 weeks before middle cerebral artery (MCA) occlusion. Infarct volume was measured at 48 h post stroke. Neurological deficits were assessed at 2 h, 24 h and 48 h post stroke. Results showed that infarct volume in rats which received saline and simvastatin was 32.5 +/- 9.3% (mean +/- SD) and 18.7 +/- 6.5%, respectively. The infarct volume in the simvastatin group was significantly smaller than in the controls (P < 0.002). Treatment with simvastatin also improved neurological deficits and reduced brain edema significantly (P < 0.05). In the second series, we studied if simvastatin can improve microvascular reperfusions after ischemia. Perfusion deficits were detected at 8 h post stroke using Evens blue dye. Neurological deficits were assessed at 2 h and 8 h post stroke. Results showed that perfusion deficit in saline and simvastatin-treated groups were 58.7 +/- 8.7% and 23.4 +/- 7.5%, respectively. The perfusion deficit in simvastatin-treated group was decreased 61% (P < 0.01). These studies thus suggest that simvastatin is a protective agent in ischemic brain injury and this protective effect may be partially due to its action in the improvement of microvascular reperfusion. PMID- 15823247 TI - Long-lasting synapse formation in cultured rat hippocampal neurons after repeated PKA activation. AB - Recently, we reported that the repeated activation of cyclic-AMP-dependent protein kinase (PKA) in the rat hippocampus under tissue culture conditions induced the enhancement of excitatory postsynaptic potential (EPSP), which lasted more than 2 weeks and was accompanied by the formation of morphologically identifiable synapses. Here we examined whether an equivalent synapse formation is induced in dissociated cell cultures of rat hippocampal neurons. Brief (15 min) application of Sp-cAMPS (a membrane-permeable analog of cyclic AMP) induced an increase in the number of synaptic sites (identified by the apposition of immunocytochemically labeled pre- and postsynaptic structures). There were two types of increase: a short-lasting one that lasted less than 24 h after a single application of Sp-cAMPS, and a long-lasting one that lasted more than 2 weeks after repeated applications. The long-lasting increase in synaptic sites was dependent on the time and interval of application and was suppressed by Rp-cAMPS (a PKA inhibitor). The synapses were judged to be active based on the endocytosis of FM1-43, a fluorescent dye. Electron microscopy confirmed the increase in the number of synaptic ultrastructures. The present results show that the synaptogenesis induced by repeated PKA activation is reproducible in a neuronal network that is reconstituted under dissociated cell culture conditions. This experimental system, together with the synaptogenesis in the slice culture system described previously, serves as a good in vitro model for the analysis of the process of conversion from short-lasting plasticity (lasting for hours) into a long-lasting one (lasting for days-weeks). PMID- 15823248 TI - Effect of protein malnutrition on redox state of the hippocampus of rat. AB - The protein malnutrition is a worldwide problem, affecting mainly newborns and children of developing countries. This deficiency reaches the brain in the most critical period of the development. Various consequences are related to this insult, such as memory disturbance, learning, and behavioral impairment. Protein content of the diet plays an important role on antioxidant mechanisms. This study observed the effects of protein malnutrition on rat hippocampus redox state. Wistar rats were separate in four groups, receiving different diets: first group with 25% casein, protein deficient group with 8% casein, and the same two groups supplemented with methionine (0.15%). Diets were isocaloric and were administered since the prenatal period up to the sacrifice. Rats were decapitated at 21 or 75 days old and hippocampus were isolated for measuring the lipoperoxidation by TBARS method, protein oxidative damage by carbonyl (DNPH) levels, and the activities of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). There was significant alterations in the activities of the enzyme SOD, lipoperoxidation, and protein oxidation in hippocampus of 21 and 75 day-old rats fed with 25% of protein with methionine and the groups fed with low levels of protein (8%) both supplemented or not with methionine. Our data suggest that both the content of protein in the diet and the essential amino acid methionine may alter the antioxidant system and the redox state of the brain. PMID- 15823249 TI - Synaptic plasticity in the dy2J mouse model of laminin alpha2-deficient congenital muscular dystrophy. AB - Laminin alpha2-deficient congenital muscular dystrophy is a debilitating disease affecting both muscle and neural tissue as a result of mutations in the LAMA2 gene. It presents at or soon after birth with muscle weakness and is further characterised by clinical central nervous system involvement. Laminin alpha2 is part of the extracellular matrix, linked to the cellular cystoskeleton via dystroglycan which is an integral part of the dystrophin-glycoprotein complex (DGC). We examined both short- and long-term synaptic plasticity in the C57BL6J/dy(2J) mouse, an animal model of laminin alpha2 deficient congenital muscular dystrophy. Using a cerebellar slice preparation, we show that the pre synaptically mediated paired-pulse facilitation (PPF) was no different between dy(2J) and littermate controls. Approximately half (7/12) the dy(2J) Purkinje cells displayed a blunted LTD compared to littermate controls, and one third (4/12) of dy(2J) Purkinje cells displayed LTP. This study demonstrates that a defective laminin alpha2 causes a disruption in long-term synaptic plasticity at the Purkinje cell-parallel fibre synapse. PMID- 15823250 TI - Temporary focal ischemia in the mouse: technical aspects and patterns of Fluoro Jade evident neurodegeneration. AB - Animal models of cerebral infarction are crucial to understanding the mechanisms of neuronal survival following ischemic brain injury and to the development of therapeutic interventions for victims of all types of stroke. Rodents have been used extensively in such research. One rodent model of stroke utilizes either permanent or temporary occlusion of the middle cerebral artery (MCAO) to produce ischemia. Since the development of an endovascular method for this was published in 1989, MCAO has been applied commonly to the rat, and often paired with 2, 3, 5 triphenyltetrazolium chloride (TTC) staining for stroke volume measurement. Meanwhile, advances in the ability to genetically alter mice have allowed exciting lines of research into ischemia. Because of technical demands and issues with survival, relatively few laboratories have investigated the MCAO method in the mouse. Our present work utilizes a mouse middle cerebral occlusion (MCAO) model of embolic stroke to study neuronal degeneration following temporary focal cerebral ischemia. C57Bl/6J mice were used to examine the exact effects of MCAO using Fluoro-Jade, a marker of neurodegeneration that allows observation of specific brain regions and cells destined to die. A time course of escalating neuronal degeneration from 10 min to 7 days following MCAO was established. Technical aspects of this popular method for transient focal ischemia as it applies to the mouse are discussed. PMID- 15823251 TI - Interaction of GABA and glutamate in the horizontal limb of diagonal band of Broca (hDB): role in cardiovascular responses. AB - The diagonal band of Broca (DBB) is a part of the limbic system that consists of two parts: the vertical limb (vDB) and the horizontal limb (hDB). It has been shown that microinjection of glutamate into the hDB of the anesthetized rat elicited depressor responses. We have previously shown that microinjection of AP5 (an NMDA receptor antagonist, 2.5 mM, 50 nl) and CNQX (an AMPA receptor antagonist, 1 mM, 50 nl) caused no significant changes in the blood pressure and heart rate. Microinjection of bicuculline (BMI: a GABA(A) receptor antagonist, 1 mM, 50 nl) resulted in the increase of both the blood pressure and heart rate. In this study, we investigated the possible interaction of the GABAergic and glutaminergic systems of the hDB by coinjection of the antagonists of both systems. Experiments were performed on the 24 urethane-anesthetized rats. Repeated measures ANOVA was used for data analysis. Our results showed that coinjection of 50 nl of 1 mM of BMI and 2.5 mM of AP5 significantly (ANOVA, P < 0. 01) decreased the pressor effects of BMI. Also, coinjection of 50 nl of BMI (1 mM) and CNQX (1 mM) significantly (ANOVA, P < 0.01) decreased the pressor effects of BMI. Coinjection of the previous amounts of BMI and both of the glutamate receptor antagonists also produced the same results. These results showed that the cardiovascular effects of blockade of GABAergic inhibition in the hDB are dependent on the activation of local NMDA and non-NMDA receptors of glutamate. A possible interpretation of the results is that, the GABAergic neurons inhibit the glutaminergic neurons. PMID- 15823252 TI - The effect of endoneurial nerve growth factor on calcitonin gene-related peptide expression in primary sensory neurons. AB - Recent findings indicate that calcitonin gene-related peptide (CGRP) is involved in neuropathic pain, this peptide being up-regulated in a small population of large- and medium-sized primary sensory neurons after peripheral nerve injury. In adult animals, the expression of CGRP is regulated by nerve growth factor (NGF). After nerve injury, NGF is up-regulated at the injury site for several weeks, and this up-regulation contributes to the onset of neuropathic pain. Using immunohistochemistry, we investigated the time course of the effect of an endoneurial injection of NGF on the expression of CGRP in primary sensory neurons. NGF increased the percentage of medium- to large-sized DRG neuron profiles expressing CGRP, did not modify the percentage of small-sized neurons expressing CGRP, and increased CGRP expression in the laminae III and IV of the dorsal horn. The effects of NGF were evident as soon as 1 day after endoneurial injection, and lasted for 5 days. Ten days after the injection of NGF, the patterns of CGRP expression in the DRG were normal, whereas a slight decrease in CGRP content was observed in the dorsal horn. The injection of vehicle did not produce any change on CGRP expression in primary sensory neurons. These results suggest that endoneurial NGF is responsible for the increase in CGRP expression in some large-sized neurons and their central processes observed after nerve injury in animal models of neuropathic pain. Our findings contribute to the understanding of the role of NGF in neuropathic pain. PMID- 15823253 TI - Altered quinpirole-induced local cerebral glucose utilization in anterior cortical regions in rats after sensitization to quinpirole. AB - Dopaminergic psychostimulants produce behavioral responses of greater magnitude with repeated, intermittent administration, than a single, acute dose, a phenomenon known as "sensitization." Most studies of sensitization have focused on the "motive circuit"; however, some additional anterior cortical regions also appear to be affected. In this study, alterations in regional neuronal activity in anterior cortical brain areas produced by quinpirole, a D(2)/D(3) agonist, were assessed on the basis of local cerebral glucose utilization (LCGU) using the [(14)C]-2-deoxyglucose (2-DG) method. Adult, male Long-Evans rats (180-200 g, n = 7-9/group) were subjected to ten injections of quinpirole (0.5 mg/kg, s.c.) administered every third day; controls and drug-naive rats received saline. Locomotor activity was quantitated after injections one and ten to confirm sensitization. The 2-DG procedure was initiated 60 min after an 11th injection in freely moving rats. LCGU was determined in 11 anterior cortical brain regions by quantitative autoradiography. In drug-naive rats, quinpirole decreased LCGU in the cingulate cortex-area 3 (-16%) and infralimbic cortex (-16%). In sensitized rats, quinpirole decreased LCGU in the cingulate cortex-area 1 (-19%), frontal cortex-area 3 (-19%), lateral orbital cortex (-18%), medial/ventral orbital cortex (-17%), and parietal cortex (-17%) as well as in the cingulate cortex-area 3 (-19%) and infralimbic cortex (-20%); (all P < 0.05 v. control). This suggests that decreased neuronal activity in the cingulate cortex-area 1, frontal cortex area 3, lateral orbital cortex, medial/ventral orbital cortex, and parietal cortex, in addition to altered activity in the motive circuit, may underlie the augmented behavioral response to quinpirole in sensitized animals. PMID- 15823254 TI - Unilateral and bilateral projections from cortical cells to the inferior colliculus in guinea pigs. AB - Auditory cortex projects directly and bilaterally to the inferior colliculus (IC). We used multiple fluorescent retrograde tracers to determine whether individual cortical cells project to both the left and right IC. Injection of different tracers into each IC labeled many cells in a sheet that extended throughout much of temporal cortex in both hemispheres. Most cells contained a single tracer, with the majority of these labeled from the ipsilateral IC. Numerous double-labeled cells were observed throughout the same areas of temporal cortex. The double-labeled cells form a small percentage of the cortical cells that project to the ipsilateral IC (6.1% on average) and a much larger percentage of the cells that project to the contralateral IC (46.4% on average). Unilaterally projecting cells are well positioned to have effects limited to one IC, whereas bilaterally projecting cells are likely to have a broader influence and may coordinate activity on the two sides of the midbrain. PMID- 15823255 TI - Modifications of 5-HT4 receptor expression in rat brain during memory consolidation. AB - Pharmacological evidence indicates a specific role of 5-HT(4) receptors on memory function. These receptors are members of G-protein-coupled 7-transmembrane domain receptor superfamily, are positively coupled to adenylyl cyclase, and are heterogeneously located in some structures important for memory, such as the hippocampus and cortical regions. To further clarify 5-HT(4) receptors' role in memory, the expression of these receptors in passive (P3) untrained and autoshaping (A3) trained (3 sessions) adult (3 months) and old (P9 or A9; 9 months) male rats was determined by autoradiography. Adult trained (A3) rats showed a better memory respect to old trained (A9). Using [(3)H] GR113808 as ligand (0.2 nM specific activity 81 Ci/mmol) for 5-HT(4) receptor expression, 29 brain areas were analyzed, 16 areas of A3 and 17 of A9 animals displayed significant changes. The medial mammillary nucleus of A3 group showed diminished 5-HT(4) receptor expression, and in other 15 brain areas of A3 or 10 of A9 animals, 5-HT(4) receptors were increased. Thus, for A3 rats, 5-HT(4) receptors were augmented in olfactory lobule, caudate putamen, fundus striatum, CA2, retrosplenial, frontal, temporal, occipital, and cingulate cortex. Also, 5-HT(4) receptors were increased in olfactory tubercule, hippocampal CA1, parietal, piriform, and cingulate cortex of A9. However, hippocampal CA2 and CA3 areas, and frontal, parietal, and temporal cortex of A9 rats, expressed less 5-HT(4) receptors. These findings suggest that serotonergic activity, via 5-HT(4) receptors in hippocampal, striatum, and cortical areas, mediates memory function and provides further evidence for a complex and regionally specific regulation over 5-HT receptor expression during memory formation. PMID- 15823256 TI - Fluctuations of the spontaneous discharge in the posterior insular cortex neurons are associated with changes in the cardiovascular system in rats. AB - Extracellular neuronal responses were recorded from the posterior insular cortex. In three of 20 neurons, fluctuations in their spontaneous discharge were observed during recording without stimulation. In these recordings, fluctuations were also observed in arterial blood pressure (BP) and heart rate (HR) recorded simultaneously. For 3 neurons, the relationships among the fluctuations in the spontaneous discharge of the insular cortex neuron (INSneu), mean arterial pressure (MAP), and HR were analyzed using Pearson's correlation coefficient (r). In unit A, there was a negative correlation between INSneu and MAP (r = -0.30). The r between INSneu and HR was -0.02. In unit B, the data were divided into two groups according to HR (HR < 400 and HR > or = 400). The differential results in the correlations for INSneu-MAP were obtained in the two groups (r = -0.2, HR < 400; r = 0.51, HR > or = 400). For unit C, the INSneu was positively correlated with BP (r = 0.31) and HR (r = 0.36). From the correlation analysis concerning the time, changes in INSneu seem to precede (or delay) changes in BP. These results showed that fluctuations in neuronal activity in the posterior insular cortex are positively or negatively correlated with BP (or HR). The data suggest that some of the neurons in the posterior insular cortex may play a role in the homeostatic (and/or regulatory) control of the autonomic system. PMID- 15823257 TI - AMPA receptor stimulation mediates the antidepressant-like effect of a group II metabotropic glutamate receptor antagonist. AB - (1R,2R,3R,5R,6R)-2-amino-3-(3,4-dichlorobenzyloxy)-6-fluorobicyclo[3.1.0]hexane 2,6-dicarboxylic acid (MGS0039), a selective group II metabotropic glutamate receptor (mGluR) antagonist, exhibits antidepressant-like activities in rodent models. In the present studies, to clarify the involvement of alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor activation in exhibition of the antidepressant-like properties of MGS0039, we examined the effect of an AMPA receptor antagonist, 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline (NBQX), on the antidepressant-like effect of MGS0039 in the mouse tail suspension test. We also examined the effects of NBQX on increased serotonin release after treatment with MGS0039 in the rat medial prefrontal cortex (mPFC) using in vivo microdialysis evaluation. In the tail suspension test, MGS0039 (0.3-3 mg/kg, i.p.) treatment dose-dependently and significantly reduced immobility time. Pretreatment with NBQX (10 mg/kg, s.c.) significantly prevented the antidepressant-like effect of MGS0039 in the tail suspension test, while NBQX itself had no effect on immobility time. In the microdialysis evaluation, administration of MGS0039 (10 mg/kg, i.p.) significantly increased serotonin levels in mPFC in freely moving rats, while NBQX (1 mg/kg, i.p.) itself had no effect on serotonin release in this region. Pretreatment with NBQX significantly attenuated the increase in serotonin release by MGS0039. These findings suggest that stimulation of postsynaptic AMPA receptors plays a role in mediating the pharmacological effects of MGS0039. PMID- 15823258 TI - Repetition of mGluR-dependent LTD causes slowly developing persistent reduction in synaptic strength accompanied by synapse elimination. AB - Synaptic plasticity, the cellular basis of memory, operates in a bidirectional manner. LTP (long-term potentiation) is followed by structural changes that may lead to the formation of new synapses. However, little is known whether LTD (long term depression) is followed by morphological changes. Here we show that the repetitive induction of metabotropic glutamate receptor (mGluR)-dependent LTD in stable cultures of rat hippocampal slices led to a slowly developing persistent (ranging over weeks) reduction in synaptic strength that was accompanied by the loss of synaptic structures. LTD was induced pharmacologically 1-3 times at 24-h intervals by applying aseptically ACPD (1-aminocyclopentane-1,3-dicarboxylic acid), an agonist of group I/II mGluR, and APV (2-amino-5-phosphonovalerate), an antagonist of the NMDA (N-methyl-D-aspartate) receptor. One ACPD/APV application induced LTD that lasted less than 24 h. After three LTD inductions, however, a gradual attenuation of the fEPSP (field excitatory postsynaptic potential) amplitude and a decrease in the number of pre- and postsynaptic structures were observed. The blockade of LTD by an mGluR antagonist or a protein phosphatase 2B inhibitor abolished the development of the synaptic attenuation. In contrast to our previous finding that the repetitive LTP induction triggered a slowly developing persistent synaptic enhancement, the incremental and decremental forms of synaptic plasticity appeared to occur symmetrically not only on the minutes hours time order but also on the days-weeks time order. PMID- 15823259 TI - Effect of systemic and intrathecal gabapentin on allodynia in a new rat model of postherpetic neuralgia. AB - Patients with postherpetic neuralgia often have an increased sensitivity to a tactile stimulus but impaired thermal sensitivity in the same affected dermatomes. We recently found that depletion of capsaicin-sensitive afferents by systemic treatment with a potent TRPV1 agonist, resiniferotoxin, in adult rats produces long-lasting paradoxical changes in mechanical and thermal sensitivities, which resemble the unique clinical features of postherpetic neuralgia. The anticonvulsant gabapentin is effective in reducing the subjective pain score in patients with postherpetic neuralgia. In this study, we quantified the potential effect of systemic and intrathecal gabapentin on tactile allodynia induced by resiniferotoxin in rats. Intraperitoneal injection of 200 microg/kg resiniferotoxin produced a rapid and sustained increase in the paw withdrawal latency to a radiant heat stimulus. Profound tactile allodynia developed in all the resiniferotoxin-treated rats within 3 weeks. Intraperitoneal injection of 30 60 mg/kg of gabapentin in resiniferotoxin-treated rats significantly increased the withdrawal threshold in response to von Frey filaments. Furthermore, intrathecal administration of 10-30 microg of gabapentin also produced a significant effect on the mechanical withdrawal threshold in all resiniferotoxin treated rats. These data provide complementary new information that gabapentin administered systemically and spinally can effectively relieve tactile allodynia in this animal model of postherpetic neuralgia. PMID- 15823261 TI - Preventing cervical cancer in low-resource settings: building a case for the possible. PMID- 15823260 TI - Ketogenic diet prevents clusterin accumulation induced by kainic acid in the hippocampus of male ICR mice. AB - We investigated the effect of ketogenic diet (KD) on clusterin accumulation in the kainic acid (KA)-induced seizure model. Two days after KA administration, strong clusterin-like immunoreactivity (IR) was detected in the hippocampus in the normal diet (ND)-fed mice. But in the KD-fed mice, few clusterin-like IR was detected. These results indicate that KD has neuroprotective effects throughout diminishing nuclear clusterin accumulation that is involved in caspase-3 independent cell death mechanism. PMID- 15823262 TI - Experience using cryotherapy for treatment of cervical precancerous lesions in low-resource settings. AB - Cervical cancer can be successfully prevented if timely identification of precancerous lesions is followed by effective treatment. In many developing countries, treatment of precancer is neglected because therapeutic services are unavailable, inaccessible, inappropriate, or inadequately linked to screening services. One of the main focuses of the Alliance for Cervical Cancer Prevention (ACCP) has been to ensure that safe and effective methods of treatment for precancer are both available and accessible to women who need them. Cryotherapy, in use for the past 40 years, is a relatively simple, safe, effective, acceptable, and appropriate outpatient procedure for the treatment of precancer. ACCP studies conducted in more than a dozen developing countries show that cryotherapy for precancer can be performed safely and effectively as an outpatient procedure at all levels of health facilities by trained and competent midlevel providers, thus increasing availability and accessibility to precancer treatment services. PMID- 15823263 TI - Delivering cervical cancer prevention services in low-resource settings. AB - The goals of any cervical cancer prevention program should be threefold: to achieve high coverage of the population at risk, to screen women with an accurate test as part of high-quality services, and to ensure that women with positive test results are properly managed. This article focuses on the experiences of the Alliance for Cervical Cancer Prevention (ACCP) in delivery of screening and treatment services as part of cervical cancer prevention projects in Africa, Latin America, and Asia. Research and experience show that cervical cancer can be prevented when strategies and services are well planned and well managed and when attention is paid to program monitoring and evaluation. Coordination of program components, reduction of the number of visits, improvement of service quality, and flexibility in how services are delivered are all essential features of an effective service. PMID- 15823264 TI - Training for cervical cancer prevention programs in low-resource settings: focus on visual inspection with acetic acid and cryotherapy. AB - The modern approach to cervical cancer prevention, characterized by use of cytology and multiple visits for diagnosis and treatment, has frequently proven challenging and unworkable in low-resource settings. Because of this, the Alliance for Cervical Cancer Prevention (ACCP) has made it a priority to investigate and assess alternative approaches, particularly the use of visual screening methods, such as visual inspection with acetic acid (VIA) and visual inspection with Lugol's iodine (VILI), for precancer and cancer detection and the use of cryotherapy as a precancer treatment method. As a result of ACCP experience in providing training to nurses and doctors in these techniques, it is now widely agreed that training should be competency based, combining both didactic and hands-on approaches, and should be done in a clinical setting that resembles the service-delivery conditions at the program site. This article reviews ACCP experiences and perceptions about the essentials of training in visual inspection and cryotherapy and presents some lessons learned with regard to training in these techniques in low-resource settings. PMID- 15823265 TI - Involving the community in cervical cancer prevention programs. AB - Underutilization of cervical cancer prevention services by women in the high-risk age group of 30-60 years can be attributed to health service factors (such as poor availability, poor accessibility, and poor quality of care provided), to women's lack of information, and to cultural and behavioral barriers. The Alliance for Cervical Cancer Prevention (ACCP) partners have been working to identify effective ways to increase women's voluntary participation in prevention programs by testing strategies of community involvement in developing countries. The ACCP experiences include developing community partnerships to listen to and learn from the community, thereby enhancing appropriateness of services; developing culturally appropriate messages and educational materials; making access to high-quality screening services easier; and identifying effective ways to encourage women and their partners to complete diagnosis and treatment regimens. Cervical cancer prevention programs that use these strategies are more likely to increase demand, ensure follow-through for treatment, and ultimately reduce disease burden. PMID- 15823266 TI - A critical assessment of screening methods for cervical neoplasia. AB - The objective of cervical cancer screening is to reduce cervical cancer incidence and mortality by detecting and treating precancerous lesions. Conventional cytology is the most widely used cervical cancer screening test. Although cytology has been effective in reducing the incidence of and mortality from cervical cancer in developed countries in both opportunistic and--more dramatically--organized national programs, it has been less successful and largely ineffective in reducing disease burden in low-resource settings where it has been implemented. Liquid-based cytology, testing for infection with oncogenic types of human papillomaviruses, visual inspection with 3-5% acetic acid, magnified visual inspection with acetic acid, and visual inspection with Lugol's iodine have been evaluated as alternative tests. Their test characteristics, and the applications and limitations in screening, are discussed with an emphasis on the work of the Alliance for Cervical Cancer Prevention over the past 5 years. PMID- 15823267 TI - Advocating for cervical cancer prevention. AB - Cervical cancer is a significant health problem among women in developing countries. Contributing to the cervical cancer health burden in many countries is a lack of understanding and political will to address the problem. Broad-based advocacy efforts that draw on research and program findings from developing country settings are key to gaining program and policy support, as are cost effectiveness analyses based on these findings. The Alliance for Cervical Cancer Prevention (ACCP) has undertaken advocacy efforts at the international, regional, national, and local levels to raise awareness and understanding of the problem (and workable solutions), galvanize funders and governments to take action, and engage local stakeholders in ensuring program success. ACCP experience demonstrates the role that evidence-based advocacy efforts play in the ultimate success of cervical cancer prevention programs, particularly when new screening and treatment approaches-and, ultimately, radically new approaches such as a human papillomavirus vaccine-are available. PMID- 15823268 TI - Preventing cervical cancer in low-resource settings: how far have we come and what does the future hold? AB - The Alliance for Cervical Cancer Prevention (ACCP) came together in 1999 to answer key research questions and to advocate for greater global and national interest in reducing the heavy burden of morbidity and mortality caused by this preventable disease. Visual inspection with acetic acid (VIA), visual inspection with Lugol's iodine (VILI), and human papillomavirus (HPV) tests have been shown to be viable alternatives to traditional cytology. ACCP experience confirmed that cryotherapy is a safe and effective method that is acceptable to women and can be delivered by a range of health providers, including nonphysicians. Programs can maximize coverage by accommodating local needs and involving community leaders and women in planning and implementation. Advocacy efforts have led to significant policy changes and galvanized support for cervical cancer prevention. Despite the prospect of new HPV vaccines, screening will be needed for at least the next 30-40 years. Our experience has shown that with creativity, flexibility, and well-focused use of resources, the inequitable burden of cervical cancer borne by women in poor countries can be sharply reduced. PMID- 15823269 TI - Tracking atherosclerosis regression: a clinical tool in preventive cardiology. AB - Progression of coronary artery disease was initially evaluated using quantitative coronary angiography with ensuing evidence indicating a strong relationship to adverse cardiovascular outcomes. Since then, several other atherosclerosis imaging techniques have emerged as new tools in cardiovascular medicine to evaluate the effectiveness of preventive therapies through serial monitoring of changes in atherosclerosis burden. Conducting large randomized trials to test new approaches for the medical management of atherosclerosis, with the goal of showing a reduction in event rates, may often be impractical in an era of cost containment and reduced societal resources. Recent evidence has unfolded that investigates alternative ways of assessing therapeutic results such as the attainment of surrogate goals with substantial outcome relevance. Atherosclerosis imaging modalities such as coronary computed tomography, carotid ultrasound, cardiovascular magnetic resonance imaging, and intravascular ultrasound each possess specific imaging abilities and inter-test characteristics that enable their serial use as intermediate endpoints in clinical trials and, increasingly, in individual patient management. The current review focuses on the application of these modalities as emerging tools in cardiovascular prevention. PMID- 15823270 TI - Disruption of tumor necrosis factor-alpha gene diminishes the development of atherosclerosis in ApoE-deficient mice. AB - Inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) have been implicated in atherogenesis. However, the precise role of TNF-alpha in atherogenesis is still unclear. To examine the effect of TNF-alpha on atherogenesis, we generated compound-deficient mice in apolipoprotein E (apoE) and TNF-alpha (apoE-/-/TNF-alpha-/-) and compared them with apoE-/- mice. Although serum total cholesterol levels were markedly elevated in both apoE-/ /TNF-alpha-/- and apoE-/- mice compared to wild-type mice, no differences were observed between apoE-/-/TNF-alpha-/- and apoE-/- mice. The atherosclerotic plaque area in the aortic luminal surface of apoE-/-/TNF-alpha-/- mice (n=8, 3.1+/-0.4%) was significantly smaller than that of apoE-/- mice (n=7, 4.7+/-0.4%, p<0.001) despite the lack of difference in serum cholesterol levels. The atherosclerotic lesion size in the aortic sinus of apoE-/-/TNF-alpha-/- mice (n=10, 5.1+/-0.3 x 10(5)microm(2)) was also significantly smaller than that of apoE-/- mice (n=11, 7.0+/-0.3 x 10(5)microm(2), p<0.0001). RT-PCR analysis indicated that the expression levels of intercellular adhesion molecule-1 (ICAM 1), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1) were significantly higher in apoE-/- than apoE-/-/TNF-alpha-/- mice. Macrophages from apoE(-/-) mice showed higher uptake level of oxidized LDL and increased expression level of scavenger receptor class A (SRA) compared to those from apoE-/-/TNF-alpha-/- mice. These results indicate that TNF-alpha plays an atherogenic role by upregulating the expressions of ICAM-1, VCAM-1 and MCP-1 in the vascular wall, and by inducing SRA expression and oxidized LDL uptake in macrophages. PMID- 15823271 TI - Tocotrienols reduce 25-hydroxycholesterol-induced monocyte-endothelial cell interaction by inhibiting the surface expression of adhesion molecules. AB - The migration of circulating monocytes into the subendothelial space occurs through the expressing of some adhesion molecules on endothelial cells. In the present study, using human aortic endothelial cells (HAECs), we investigated whether a model compound for oxysterols, 25-hydroxycholesterol, can enhance the monocyte adherence to HAECs exposed to 25-hydroxycholesterol via increasing expression of vascular cell adhesion molecule-1 (VCAM-1). We also aimed to determine the in vitro effects of tocotrienols on the enhanced interaction between monocytes and endothelial cells. We found that 25-hydroxycholesterol enhances surface expression determined by ELISA, induces VCAM-1 mRNA expression by real time-PCR, and stimulates adhesiveness of HAECs to U937 monocytic cells in a dose-dependent fashion. The combination treatment with anti-VCAM-1 and anti CD11b monoclonal antibodies significantly reduced the monocyte adherence to 25 hydroxycholesterol-stimulated HAECs. Compared to alpha-tocopherol, tocotrienols displayed a more profound inhibitory effect on adhesion molecule expression and monocytic cell adherence. We observed that delta-tocotrienol exerted a most profound inhibitory action on monocytic cell adherence when compared to alpha tocopherol and alpha-, beta-, and gamma-tocotrienols. Tocotrienols accumulated in HAECs to levels approximately 25-95-fold greater than that of alpha-tocopherol. In conclusion, these results indicate that a model compound 25-hydroxycholesterol can enhance the interaction between monocytes and HAECs, and that tocotrienols had a profound inhibitory effect on monocytic cell adherence to HAECs relative to alpha-tocopherol via inhibiting the VCAM-1 expression. These superior inhibitory effects of tocotrienols may be dependent on their intracellular accumulation. PMID- 15823272 TI - Inhibition of cytokine-induced IkappaB kinase activation as a mechanism contributing to the anti-atherogenic activity of tilianin in hyperlipidemic mice. AB - Tilianin has been shown to down-regulate TNF-alpha induced expression of vascular cell adhesion molecules in endothelial cells. In this study, we examined the anti atherogenic effects and molecular mechanism of tilianin in vitro and in vivo. Male low-density lipoprotein receptor null mice (Ldlr-/-) fed a high cholesterol diet showed significant increases in the size of atherosclerotic lesions, as well as increased plasma levels of total cholesterol, triglycerides, and the pro inflammatory cytokines TNF-alpha and IL-1beta, when compared with Ldlr-/- mice fed a normal diet. Mice fed the high cholesterol diet supplemented with tilianin showed significantly reduced lesion sizes and reductions in cytokine levels, without significant changes in serum cholesterol levels. Primary cultured peritoneal macrophages from Ldlr-/- mice showed increased level of TNF-alpha andIL-1beta mRNA in response to treatment with lipopolysaccharide; these increases were inhibited by co-treatment with tilianin. Moreover, tilianin inhibited NF-kappaB activation, as determined by electrophoretic mobility shift and NF-kappaB promoter assays. Upstream of NF-kappaB activation, tilianin inhibited IkappaB kinase activation and the subsequent phosphorylation and degradation of IkappaBalpha protein. These results suggest that tilianin ameliorates atherosclerosis by inhibiting the production of the NF-kappaB dependent pro-inflammatory cytokines, TNF-alpha and IL-1beta, via the inhibition of IkappaB kinase activity. PMID- 15823273 TI - Upstream regulation of matrix metalloproteinase by EMMPRIN; extracellular matrix metalloproteinase inducer in advanced atherosclerotic plaque. AB - From experimental and clinical studies it is known that matrix conservation and degradation by matrix metalloproteinases (MMPs) plays a major role in plaque progression and destabilization with related onset of acute vascular events such as acute coronary syndromes or cerebrovascular accidents. Recently, extracellular MMPs inducer (EMMPRIN) has been reported to induce and activate the expression of MMPs in myocardium and plays an important role in the ventricular remodeling in human heart failure. Similarly to heart failure myocardium, EMMPRIN may be expressed in human atheroma and play a role in the extracellular matrix (ECM) remodeling and atherogenic cell differentiation. This study was designed to investigate the possible biological role of EMMPRIN in human atheroma. Immunohistochemical analysis for MMPs and EMMPRIN was performed on human carotid endarterectomy specimens and control aortas. EMMPRIN showed significant immunoreactivity in human atherosclerotic carotid lesions, and was colocalized with macrophage/monocyte infiltrates in atherosclerotic intima, plaque itself and vascular smooth muscle cells (VSMCs). Zymography and Western blot analysis revealed EMMPRIN expression in the carotid atheromas, but not in the control aortas. Human bone marrow monocytes, which were cultured with atherogenic proinflammatory cytokine stimulation revealed increased EMMPRIN and MMPs expressions. ECM remodeling is under the control of induction and inhibition of matrix degrading protease and the novel MMP inducer, EMMPRIN may play a role in influx and differentiation of monocytes and destabilizing atheroma. PMID- 15823274 TI - Absence of the protease inhibitor cystatin C in inflammatory cells results in larger plaque area in plaque regression of apoE-deficient mice. AB - Matrix remodelling plays an important role in regulating plaque stability. Cystatin C, an inhibitor of the elastin-degrading cysteine proteases of the cathepsin family, is believed to be one of the key protease inhibitors in this process. The aim of the present study was to investigate the role of leukocyte specific cystatin C expression under conditions that favour plaque regression. Apolipoprotein E-deficient mice (apoE-/-) were given a Western-type diet 15 weeks prior transplantation with bone marrow from mice lacking cystatin C (cysC-/-) or cystatin C positive (cysC+/+) mice, in both cases apoE+/+ to create conditions favouring plaque regression. Transplantations were verified with PCR and Western analyses. Transplanted mice showed a 70% decrease in lipid content and reduction in plaque area compared to baseline ApoE-/- mice, demonstrating plaque regression due to apoE expression in macrophages. apoE-/- mice transplanted with cysC-/- bone marrow were then compared to mice transplanted with cysC+/+ bone marrow. Mice receiving cysC-/- bone marrow had a 30% larger plaque area, despite absence of significant differences in plasma cholesterol and lipid contents in plaque. Unexpectedly, mice transplanted with cystatin C-deficient bone marrow cells had increased elastin and collagen content in lesions. These observations suggest that leukocyte-specific expression of cystatin C is actively involved in matrix remodelling associated with plaque regression. PMID- 15823275 TI - Paraoxonase 1 (PON1) is present in postprandial chylomicrons. AB - Paraoxonase 1 (PON1) is an esterase, associated in serum with high density lipoprotein (HDL). As diet affects serum PON1 activity, we questioned whether PON1 is also carried by postprandial chylomicrons. Chylomicrons were isolated by ultracentrifugation from plasma of 10 healthy men, 3h after the consumption of a high fat, high carbohydrate meal, and were analyzed for the presence of PON1 arylesterase activity and protein. The present study shows for the first time that, in addition to the presence of PON1 mainly on HDL, postprandial chylomicrons also contain minor, but significant amount of PON1. PON1 was also present in chylomicrons derived from fasted patients with hyperchylomicronemia. In addition, PON1 was detected in very low density lipoprotein (VLDL), but not in LDL. The origin of chylomicron PON1 could be partly attributed to its transfer from HDL. Finally, this study demonstrates that postprandial chylomicrons inhibit copper ion-induced LDL oxidation, secondary to hydrolysis of lipid peroxides, a phenomenon which could be related, at least in part, to the chylomicron PON1 content. We conclude that postprandial chylomicrons contain PON1, which may function in the removal of atherogenic oxidized lipids. PMID- 15823276 TI - The molecular basis of familial hypercholesterolaemia in Turkish patients. AB - Familial hypercholesterolaemia (FH) is an autosomal dominant disorder of lipoprotein metabolism. In the majority of patients FH is caused by mutations in the gene for the low-density lipoprotein receptor (LDLR), and to date more than 700 mutations have been reported worldwide. In this study, 36 paediatric patients with a clinical diagnosis of FH (20 homozygous and 16 heterozygotes) were screened for mutations in the LDLR gene. Each exon, with intron-exon junctions, was screened by capillary fluorescent SSCP (F-SSCP) and heteroduplex analysis. Samples showing different band patterns were sequenced. Ten novel (including three frame shift small deletions or insertions) and seven known mutations were detected. A total of 37 out of the predicted 56 FH-causing alleles were identified (66.1%). No patients with the R3500Q mutation in the APOB gene were found. W556R was the most common mutation, explaining 21.4% of the predicted defective LDLR alleles. The novel sequence changes were deemed to be pathogenic if they altered a conserved amino acid (L143P, D147E, Q233H-C234G, C347G) or occurred in or close to a splice site (IVS 16+5) and were absent in DNA from 50 healthy Turkish subjects. These data confirm the genetic heterogeneity of FH in Turkey, and demonstrate the usefulness of F-SSCP for mutation detection. PMID- 15823277 TI - Matrix metalloproteinase3 and 9 gene promoter polymorphisms: joint action of two loci as a risk factor for coronary artery complicated plaques. AB - OBJECTIVE: Matrix metalloproteinases 3 and 9 (MMP3 and MMP9) are present in atherosclerotic plaques and co-operate in the degradation of the fibrous cap of the atheroma, leading to fissuring and ultimately to acute coronary thrombosis. The functional genetic polymorphisms in the promoters of MMP3 and MMP9, which lead to low- and high-transcription activity genotypes, have been shown to be associated with myocardial infarction and angiographically measured atherosclerosis individually, whereas their effects in combination are not yet known. In order to assess the two disease loci simultaneously, we investigated the association of combined low and high promoter activity genotypes with different types of coronary lesions in an autopsy cohort of 300 Caucasian males aged 33-69 years (Helsinki Sudden Death Study). METHODS: Genotyping at these loci was performed by PCR, restriction enzyme digestion and minisequencing, and areas of the coronary wall covered with atherosclerotic lesions were measured using computer-assisted morphometry. RESULTS: In analysis of covariance (ANCOVA) with age, body mass index, hypertension, diabetes, smoking and alcohol consumption as covariates, a significant interaction between the MMP3 and MMP9 genotypes was observed on area of complicated lesions (P=0.012). Men with high promoter activity genotypes for both loci had, on average, more than two times larger area of complicated lesions (250%) compared with those men who had low promoter activity genotypes (P=0.008), but these loci showed no association with myocardial infarction. CONCLUSIONS: The joint action of two susceptibility loci, rather than single MMP genes alone, and the particular combination of MMP3 and MMP9 genotypes present at these loci may contribute to heterogeneity in the presentation of atherosclerosis. PMID- 15823278 TI - Apolipoprotein A-I/C-III/A-IV SstI and apolipoprotein B XbaI polymorphisms and their association with carotid artery intima-media thickness in the Finnish population. The Cardiovascular Risk in Young Finns Study. AB - Apolipoprotein A-I/C-III/A-IV (apoA-I/C-III/A-IV) SstI and apolipoprotein B (apoB) XbaI polymorphisms have been shown to affect serum low-density lipoprotein (LDL) cholesterol concentrations in a sample of Finnish children. We studied whether these polymorphism are associated with carotid artery intima-media thickness (IMT), a marker of pre-clinical atherosclerosis, measured in the same subjects during their adulthood. A random sub-sample of 214 individuals from the "Cardiovascular Risk in Young Finns" study, for whom genotypes, cardiovascular risk factor data and carotid artery IMT measured in 2001 were available, were studied. Mean carotid IMT values increased according to the apoA-I/C-III/A-IV SstI genotype groups in the order of S1S1 (0.58+/-0.08 mm), S1S2 (0.61+/-0.08 mm), and S2S2 (0.70+/-0.16 mm, p=0.02, ANOVA). In multiple linear regression analysis after adjusting for age, sex and body mass index the mean IMT thickness among the S2 allele carriers was higher (p=0.02) compared to non-carriers. In logistic regression analysis the frequency of S2 allele carriers was higher among the high IMT group compared to the low IMT group (OR=4.02, CI: 1.68-9.61, p=0.002). No significant association between apoB XbaI polymorphism and carotid IMT was found. However, serum total and LDL cholesterol and apoB concentrations were significantly different among apoB genotype groups (p<0.001 for all traits). The apoA-I/C-III/A-IV SstI polymorphism is associated with carotid IMT in young Finns. PMID- 15823279 TI - Smoking is associated with advanced coronary atherosclerosis in youth. AB - Smoking is linked to atherosclerosis and coronary heart disease (CHD) in older adults. However, evidence that smoking affects coronary atherosclerosis in young people is incomplete. The Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Study collected arteries, blood, and other tissues from persons 15 to 34 years of age dying of external causes and autopsied in forensic laboratories. Lesions in the proximal left anterior descending coronary arteries (LAD) from 1127 subjects were graded microscopically according to the American Heart Association criteria. Among individuals with advanced lesions (Grade 4 or 5), smokers had a greater prevalence of Grade 5 lesions than non-smokers (odds ratio 9.61, 95% confidence interval 2.34-39.57), a difference suggesting that smoking accelerates the transition from Grade 4 to Grade 5 lesions. This association occurred among both men and women, and among persons with and without other CHD risk factors. The difference in qualities of advanced lesions suggests that smoking possibly accelerates the transition from Grade 4 to Grade 5 lesions by promoting thrombosis and accretion on the intimal surface of the plaque. PMID- 15823280 TI - Effect of low-density lipoprotein receptor mutation on lipoproteins and cardiovascular disease risk: a parent-offspring study. AB - Studies on the clinical consequences of different low-density lipoprotein (LDL) receptor genotypes in adult patients have yielded conflicting results. We hypothesized that children with familial hypercholesterolemia (FH) provide a better model to perform genotype-phenotype analyses than adults. We tested this hypothesis and assessed the effect of LDL receptor genotypes on lipoprotein levels and on parental risk of cardiovascular disease (CVD) in a pediatric FH cohort. We identified 75 different LDL receptor mutations in 645 children with heterozygous FH; in these children, null alleles were clearly associated with more elevated LDL cholesterol levels compared to receptor-defective mutations. Familial factors explained 50.4% of the variation in LDL cholesterol levels of this pediatric cohort compared to only 9.5% in adults. Parental CVD risk was not significantly different between carriers of null alleles and receptor-defective mutations (RR, 1.22; 95% CI, 0.76-1.95; p=0.4). The N543H/2393del9 mutation was associated with a less deteriorated lipid profile and the parents had less often CVD relative to parents with other mutations (RR, 0.39; 95% CI, 0.20-0.78; p=0.008). We could confirm that children with FH provide a better model to perform genotype-phenotype analyses. In particular, children with null alleles had significantly more elevated LDL cholesterol levels than carriers of other alleles but this was not associated with higher risk of CVD in the parents. Nonetheless, a specific LDL receptor mutation was associated with less deteriorated lipoprotein levels and a milder CVD risk. PMID- 15823281 TI - Elevated levels of oxidized low-density lipoprotein and impaired nocturnal synthesis of melatonin in patients with myocardial infarction. AB - This study was designed to investigate the relationship between nocturnal serum melatonin (MEL) levels and oxidized low-density lipoprotein (OxLDL) in patients with acute coronary syndrome (ACS). OxLDL plays a pivotal role in the development of atherosclerosis. Patients with coronary heart disease have an impaired nocturnal secretion of MEL. To date, there are no clinical human studies concerning the relationship of MEL to low-density lipoprotein (LDL) oxidation in patients with acute myocardial infarction (AMI). The study population contained 60 patients with AMI and 60 control subjects. Levels of circulating OxLDL were measured by a monoclonal antibody 4E6-based competition ELISA. Levels of circulating MEL were measured by an enzyme-immunoassay kit after chloroform extraction. Comparison of levels between AMI and controls, adjusted for age, revealed significantly higher nocturnal serum OxLDL levels (95.47+/-6.81 versus 68.35+/-4.07 U/l; p=0.004) in the AMI subjects. Nocturnal serum levels of MEL were lower in AMI than the control group (20.97+/-3.90 versus 53.19+/-7.80 pg/ml; p=0.009). Serum levels of total, high-density lipoprotein (HDL), and LDL cholesterol did not differ between the groups. Multiple regression analysis was performed on cases to study the association between AMI and serum levels of OxLDL and MEL (OR: 2.93; 95% CI, 2.89-2.98, p=0.01 and OR: 0.94; 95% CI, 0.89-0.97, p=0.02, respectively). This study demonstrates for the first time an independent association between nocturnal levels of OxLDL and MEL in patients with AMI. Additional population studies are necessary to further document these. PMID- 15823282 TI - Effects of ezetimibe on plasma lipoproteins in severely hypercholesterolemic patients treated with regular LDL-apheresis and statins. AB - Ezetimibe, a cholesterol absorption inhibitor, can be combined with statins to lower LDL-cholesterol. We evaluated additional ezetimibe (10 mg/day) in a placebo controlled, double blind, randomized cross-over study in 20 patients (age 56+/-9 years, m:f 10:10, BMI 27.5+/-4.0 kg/m(2)) suffering from severe hypercholesterolemia and CHD who were treated by statins and regular LDL apheresis. Lipoproteins (cholesterol, triglycerides, LDL-cholesterol, HDL cholesterol, VLDL-cholesterol, VLDL-triglycerides, lipoprotein(a)) were determined twice (before and during ezetimibe/placebo, each given for 5 weeks), dietary behaviour was analyzed once (3-days-protocol) during each treatment period. During ezetimibe the mean (+/-S.D.) preapheresis LDL-cholesterol concentration decreased from 159+/-26 mg/dl (4.11+/-0.67 mmol/l) to 133+/-28 mg/dl (3.44+/-0.72 mmol/l) (-16+/-11%, P<0.0001, Wilcoxon test) and the postapheresis LDL-cholesterol from 51+/-9 mg/dl (1.32+/-0.23 mmol/l) to 43+/-8 mg/dl (1.11+/-0.21 mmol/l) (-14+/-25%, P<0.05), while there was no significant change during placebo. Mean VLDL-cholesterol fell by 18+/-71% (P<0.05) during ezetimibe and was not significantly changed by placebo (+19+/-70%). Furthermore, during ezetimibe less plasma volume was treated (3725+/-1560 versus 3870+/-1549 ml, P<0.05). Ezetimibe had no effect on pre- and postapheresis triglyceride, HDL cholesterol and lipoprotein(a) levels. The effect of ezetimibe was independent of the statin dose. Dietary behaviour did not change and no side effects were observed. Thus, in patients with severe LDL-hypercholesterolemia and CHD the addition of ezetimibe to intensive lipid lowering therapy (statins and LDL apheresis) resulted in a further, clinically significant decrease of LDL cholesterol. PMID- 15823283 TI - Angiopoietin-1 and angiopoietin-2 in diabetes mellitus: relationship to VEGF, glycaemic control, endothelial damage/dysfunction and atherosclerosis. AB - BACKGROUND: microvascular complications in diabetes identify those at risk of cardiovascular disease (CVD), suggesting a link between abnormal neovascularisation and CVD. This may be related to high plasma vascular endothelial growth factor (VEGF). We hypothesised increased angiopoietins (Ang)-1 and -2 in patients with diabetes that are related to VEGF, medium-term glycaemic control, endothelial damage/dysfunction and atherosclerosis. METHODS AND PATIENTS: we measured plasma Ang-1 and Ang-2 alongside VEGF (all by ELISA) in 96 patients with type-2 diabetes mellitus (41 with and 56 without overt CVD) who were compared to 35 age- and sex-comparable healthy controls. Common carotid intima-media thickness (CC-IMT) was used to assess carotid atherosclerosis, plasma von Willebrand factor (vWf) and urine albumin:creatinine ratio (UACr) to quantify and endothelial damage/dysfunction, and HbA1c to mark medium-term hypergylcaemia. RESULTS: Ang-2 (but not Ang-1) was higher in patients with diabetes compared to controls (p<0.01), with no significant difference between patients with and without CVD. As expected, CC-IMT, UACr, HbA1c, vWf, and VEGF were also abnormal in the patients. Within the patient group alone, and in the entire cohort, VEGF and Ang-2 correlated strongly (both p<0.001) and with several other markers. However, in multivariate analysis, the only significant relationship that remained after adjustments was between VEGF and HbA1c (p<0.001). CONCLUSIONS: Angiogenic growth factor Ang-2, like VEGF, is raised in diabetes regardless of vascular disease. Both growth factors correlated with HbA1c and with each other, not with endothelial injury or atherosclerosis, but after multiple adjustment, only that between HbA1c and VEGF significant remained. VEGF is likely to have a more prominent role in diabetes than Ang-2. PMID- 15823284 TI - Low adipocyte IRS-1 protein expression is associated with an increased arterial stiffness in non-diabetic males. AB - OBJECTIVE: Low adipocyte IRS-1 protein expression is a biomarker for insulin resistance and early atherosclerosis. However, whether IRS-1 protein expression is related to systemic arterial stiffness, is unknown. METHODS AND RESULTS: Ten non-diabetic male subjects with low adipocyte IRS-1 protein expression (LIRS) were matched with 10 non-diabetic males with normal IRS-1 protein expression (NIRS). Augmentation index (AIx) and time for reflection of pulse wave (Tr) were studied with pulse wave analysis, both in the fasting state and during a euglycemic hyperinsulinemic clamp. The LIRS-group showed an increased fasting insulin concentration (fP-insulin 71+/-4 pmol/L versus 58+/-5 pmol/L; p=0.02 (mean+/-S.E.)), whereas glucose disposal rate during the clamp (8.7+/-0.8 mg/kg LBM/min versus 10.3+/-1.3 mg/kg LBM/min; n.s.) did not differ significantly. Blood pressure, lipid parameters, adiponectin, endothelin-1 and CRP concentrations were similar. However, in the basal state, AIx was increased (129+/-4% versus 116+/-2%; p<0.02) and Tr was decreased (150+/-3 ms versus 171+/ 5 ms; p<0.01), suggesting stiffer vessels in the LIRS-group. The LIRS-group exhibited an attenuated AIx response to hyperinsulinemia compared to the NIRS group. CONCLUSIONS: The data suggest that non-obese non-diabetic men with a low adipocyte IRS-1 protein expression have an increased systemic arterial stiffness. PMID- 15823285 TI - Effect of orlistat on postprandial lipemia, NMR lipoprotein subclass profiles and particle size. AB - Evidence suggests that metabolic phenomena during postprandial lipemia may be important in the pathogenesis of atherosclerosis. Both lipid concentrations and lipoprotein subclass patterns may be important cardiovascular risk modifiers. The pancreatic lipase inhibitor orlistat reduces fat absorption by 30% and is used for the treatment of overweight and obesity. We evaluated the effect of orlistat on postprandial lipemia and lipoprotein particle distribution after moderate-and high-fat meals in healthy volunteers. In this double-blind, randomized, cross over study, 10 healthy young men received orlistat 120 mg plus a high-fat meal (HFO), orlistat plus a moderate-fat meal (MFO) or placebo plus a high-fat meal (HFP). Plasma triacylglycerol, glucose, insulin, and free fatty acids were measured at baseline (fasting) and postprandially for 8h. Lipoprotein subclass profile was assessed by nuclear magnetic resonance spectroscopy. The 8h postprandial mean triacylglycerol area under the curve (AUC) was significantly lower with MFO and HFO (0.79 versus 1.33 mmol/lh) versus HFP (4.33 mmol/lh; p=0.02). Mean change in large VLDL subclass concentration during the 4-8h and mean VLDL size after 8h was significantly lower with HFO and MFO versus HFP (p<0.001). Small HDL particle concentration decreased significantly with HFP versus MFO or HFO (p<0.001). There was no significant difference in postprandial concentrations of glucose, insulin or free fatty acids on the different regimens. The lowering of postprandial triacylglycerol AUC, shorter postprandial lipemia, lower concentration of large triacylglycerol-rich particles and decrease of VLDL particle size supports the hypothesis of a less atherogenic postprandial lipoprotein profile following orlistat ingestion. PMID- 15823286 TI - LDL cholesterol and global risk stratification in referred hypertensive patients. AB - BACKGROUND: Global risk status more than BP values per se drive nowadays treatment decisions and increasing emphasis is given to the role of lipid control in hypertension (HT). However, the distribution of circulating low density lipoproteins (LDL) levels as a function of risk profile and lipid-lowering treatment in hypertensive patients is unclear. METHODS: We analysed 1196 patients (677 males, age range: 20-80 years) referred to our Hypertension Unit with treatment history and a complete dataset (systolic blood pressure levels, being on anti-hypertensive treatment or not, total and high density lipoproteins (HDL) cholesterol, smoking status, sex, age) for 10-year absolute coronary heart disease (CHD) risk stratification by National Cholesterol Education Program (NCEP)/ATP III guidelines. LDL cholesterol <25.9 mmol/L (100 mg/dL) was the target for high-risk patients (vascular diseases, diabetes, hypertension with multiple risk factors at CHD risk >20%/10 years). LDL <33.6 mmol/L (130 mg/dL) and 41.4 mmol/L (160 mg/dL) were the thresholds for intermediate (10-20%/10 years) and low (<10%/10 years) CHD risk. RESULTS: At referral, 78% of high-risk patients were above target LDL and, overall, 56% had LDL cholesterol above the desired risk-specific thresholds. Lipid-lowering treatment was prescribed in 19% in whom LDL was actually higher than the untreated group. CONCLUSIONS: LDL cholesterol was out of target in most of a large series of referred high-risk hypertensive patients and LDL levels were largely unsatisfactory even in those undergoing lipid-lowering treatment. The data show the intensive effort still needed to implement global risk-oriented prevention strategies in hypertensive populations. PMID- 15823287 TI - Assessment of arterial medial characteristics in human carotid arteries using integrated backscatter ultrasound and its histological implications. AB - Recently, ultrasound tissue characterization of the carotid arteries with an integrated backscatter (IB) analysis was shown to identify a high-risk group of atherosclerosis. To clarify whether IB ultrasound is useful in assessing arterial sclerosis as well as stiffness beta and whether IB values reflect the histological structure, we measured IB values of common carotid media in 52 subjects without coronary risk factors and in 10 patients with systemic sclerosis (SSc) in the clinical studies and 12 patients in the histological studies with a Philips Medical Systems Sonos 5500. IB values were correlated with age (r=0.69, P<0.0001), intima-media thickness (r=0.72, P<0.0001) and stiffness beta (r=0.80, P<0.0001) in the control subjects. IB values and stiffness beta in the SSc group were greater than in an age- and sex-matched control group (IB values: 9.6+/ 2.7dB versus 16.1+/-1.8dB; stiffness beta: 11.5+/-4.5 versus 20.6+/-5.6, P<0.01). IB values of the media were correlated with the elastic fragmentation index (r=0.63, P=0.029) and the collagen fiber index (r=0.59, P=0.046). Measurements of IB values of carotid media are useful for non-invasively evaluating arterial sclerosis. PMID- 15823288 TI - The relationship of molecular genetic to clinical diagnosis of familial hypercholesterolemia in a Danish population. AB - The genes encoding the LDL receptor and apoB were screened for mutations associated with familial hypercholesterolemia (FH) in 408 patients referred to the Lipid Clinic in 1995-2003. The study aimed at testing the ability of three different sets of clinical criteria to predict the results of molecular genetic analysis, and secondly test whether population-based age- and sex-specific percentiles of LDL-cholesterol offer useful supplemental information in the selection of patients for molecular genetic analysis. The patients were retrospectively categorised according to Simon Broome Register Group criteria, Make Early Diagnosis to Prevent Early Death criteria (MEDPED) and the Dutch Lipid Clinic Network criteria, and the distribution of patients was compared to the results of the molecular genetic analysis. The study illustrates a classical dilemma. Mutation detection rates (and specificities) are high only if sensitivity is very low and vice versa: to find most mutation carriers, even patients with only possible FH must be examined by molecular genetic testing leading to mutation detection rates as low as 30-40%. PMID- 15823289 TI - Association between insulin resistance and apolipoprotein B in normoglycemic Koreans. AB - BACKGROUND: Insulin resistance (IR) is associated with a significant increase in the risk of coronary artery disease (CAD). The serum apolipoprotein B and Apo A1/Apo B ratio are important markers of CAD. The aim of this study was to assess the association of the serum Apo B and Apo A1/Apo B ratio, with insulin resistance in apparently healthy normoglycemic Koreans. METHODS: From the individuals that participated in medical screening at the health promotion center of Kangbuk Samsung Hospital, between January and December 2002, a total of 7427 participants (4356 men, 3071 women) were enrolled in this study. All participants had no personal histories of diabetes, with normal fasting glucose levels. The clinical characteristics and biochemical parameters of the subjects were assessed. RESULTS: The Apo B, total cholesterol/high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein-cholesterol (LDL-C)/high-density lipoprotein-cholesterol showed positive correlations with metabolic syndrome and insulin resistance (p<0.001). The Apo A1, Apo A1/Apo B, LDL/Apo B and HDL/Apo A1 showed negative correlations with metabolic syndrome and insulin resistance (p<0.001). CONCLUSION: These data suggest that insulin resistance may be associated with the serum Apo B and Apo A1/Apo B ratio in non-diabetic, normoglycemic subjects. Thus, further study may be needed to determine whether medical intervention is inevitable or not in these type of subjects. PMID- 15823290 TI - Comparison of coronary and aortic atherosclerosis in youth from Japan and the USA. AB - This paper reports the results of the largest autopsy-based comparative study of atherosclerotic lesions between young Japanese and Americans, aged 15-34 years and autopsied between 1987 and 1995, by analyzing the data from the Japanese second nation-wide study of atherosclerosis and Pathobiological Determinants of Atherosclerosis in Youth study in the USA. In the right coronary arteries, in Japanese, fatty streaks were well established in the second decade of life with very little increase in the remaining age groups up to age of 34 years. In contrast, in American subjects, the average percentage of surface involvement of fatty streaks and raised lesions proceeded steadily with age without an obvious plateau throughout the 20-year period. The extent and prevalence of the raised lesions was much greater in Americans than in Japanese in 25-29 and 30-34-year age groups. Moreover, the rate of progression of raised lesions was much more rapid in Americans. These findings are in keeping with the fact that coronary heart disease (CHD) death rates are much higher in the USA than in Japan. In light of data showing that the risk factor profiles for CHD have become very similar between the two countries, these differences need to be explained. PMID- 15823292 TI - Association between serum C-reactive protein levels and pulse wave velocity: a population-based cross-sectional study in a general population. AB - Arterial stiffness as determined by aortic pulse wave velocity (PWV) has been shown to predict cardiovascular events in high-risk subjects such as those with hypertension or end-stage renal disease. Although it is suspected that low-grade inflammation as represented by increased C-reactive protein (CRP) plays an important role in the progression of atherosclerosis, it is not yet known whether serum CRP levels are associated with PWV. To examine the relationship between brachial-ankle PWV (baPWV) and serum CRP levels, several cardiovascular risk factors including these two markers (baPWV and CRP) were measured in 870 participants (mean age 59 years) randomly selected from a general population. Age, male gender, systolic blood pressure, heart rate, diabetes, and serum CRP levels increased with the quartiles divided by baPWV (all, p<0.01). By multiple regression analysis, age (p<0.001), systolic blood pressure (p<0.001), heart rate (p<0.001), body mass index (p<0.001), and CRP (p<0.01) were significant and independent predictors for baPWV. In conclusion, this cross sectional study has demonstrated in the general population that the arterial stiffness marker baPWV was independently correlated with serum CRP levels after adjustment for other established cardiovascular risks factors. This result suggests that baPWV may be a surrogate marker for atherosclerotic vascular damages including an inflammatory component. PMID- 15823291 TI - Interleukin-2 levels are associated with carotid artery intima-media thickness. AB - OBJECTIVE: To show that serum interleukin levels are associated with carotid intima-media thickness (IMT). BACKGROUND: Inflammation is hypothesized to play a central role in atherogenesis, and serum markers of inflammation are predictive of cardiovascular disease. Interleukin-2, a pro-inflammatory cytokine produced largely by naive CD4 T cells and Th1 (pro-inflammatory) T cells, has been found in a high proportion of carotid plaques. METHODS: High-resolution ultrasound of the carotid arteries and serum cytokine levels were measured in stroke-free participants. The mean of the maximum IMT in bilateral bifurcation, common and internal carotid artery segments was measured. Serum levels of interleukin (IL) 1beta, IL-2, IL-6, C-reactive protein, tumor necrosis factor (TNF) alpha and TNF receptors were measured using enzyme-linked immunosorbent assay. RESULTS: IL-2 levels were significantly correlated with IMT (r=0.33, P<0.0001), but other cytokines were not. Each unit increase in IL-2 was significantly associated with a mean increase in IMT of 0.18 mm (P=0.0001). After adjusting for other atherosclerotic risk factors, the association was unchanged (mean increase in IMT per unit increase IL-2=0.18 mm, P<0.0001). Each standard deviation increase in the level of IL-2 was associated with an increased risk (adjusted odds ratio 1.80, 95% CI 1.12-2.89) for an IMT> or =1.0mm (75th percentile for IMT). CONCLUSION: Serum levels of IL-2, a pro-inflammatory cytokine, are associated with carotid artery IMT, a predictor of stroke and vascular disease. Serum inflammatory markers may provide a novel marker of atherosclerotic risk, and inflammation may provide a new therapeutic target for stroke prevention. PMID- 15823293 TI - Effect of diacylglycerol on postprandial lipid metabolism in non-diabetic subjects with and without insulin resistance. AB - The effects of diacylglycerol ingestion on postprandial lipid metabolism in non diabetic subjects with and without insulin resistance were investigated. This was single dose ingestion study, in a double blind cross over manner and postprandial lipid concentrations were compared between diacylglycerol oil (DAG) and triacylglycerol oil (TAG) ingestion. The subjects were 18 male volunteers and homeostasis model assessment (HOMA-R) was used to classify them into insulin sensitive (IS, n=10, HOMA-R<2.0) and insulin resistant (IR, n=8, HOMA-R> or =2.0) groups. Fasting serum triglycerides (TG) and remnant-like particle cholesterol (RLP-C) correlated with HOMA-R and were significantly higher in the IR as compared to the IS group. Postprandial increments of TG and RLP-C after DAG ingestion were significantly lower as compared to those after TAG ingestion. In a case of TAG ingestion, their increments positively correlated with HOMA-R and were significantly higher in the IR as compared with the IS group. In contrast, their increments remained constant after DAG ingestion in both groups. In the IR group, the postprandial lipidemia were reduced after DAG ingestion to about half of those after TAG ingestion. In conclusion, DAG reduced postprandial lipidemia especially in subjects with insulin resistance and may be beneficial in preventing atherosclerosis and related diseases. PMID- 15823294 TI - Intracoronary ultrasound guided percutaneous coronary angioplasty using a drug eluting stent in a patient with pseudoxanthoma elasticum. PMID- 15823295 TI - Increased titer of antibodies to Mycoplasma pneumoniae may be associated with coronary heart disease. PMID- 15823296 TI - A novel common variant -181 G insertion in the promoter of the gene for phospholipid transfer protein. PMID- 15823298 TI - Molluscs as multidisciplinary models in environment toxicology. AB - The primary goal of this paper is to stimulate discussion and promote the use of mollusc models and multidisciplinary research approaches in the field of environmental toxicology. Molluscs are effective models because they are ubiquitous, have highly conserved control and regulatory pathways that are often homologous to vertebrate systems, and are extremely sensitive to anthropogenic inputs. We have attempted to provide initial references as an avenue into the literature and as a means for researchers to expand their thoughts to regions outside of their own area of expertise. The authors are particularly interested in developing multidisciplinary international collaborative efforts. PMID- 15823299 TI - Establishing the ecological quality status of soft-bottom mining-impacted coastal water bodies in the scope of the Water Framework Directive. AB - The aim of the present study was to check the usefulness of the benthic biotic indices proposed for application in the European water framework directive (WFD 2000/60/EC) for the ecological quality classification of coastal water bodies, together with some other benthic methodologies used in different countries of the world. The different approaches were applied in two marine ecosystems affected by the same heavy metal contamination source, coastal waters off Portman and the Mar Menor coastal lagoon, both in SE Spain. Two marine biotic indices proposed for application in the Directive (AMBI and BENTIX) were used, together with community descriptors (abundance, Shannon-Wiener diversity, Margalef's species richness, Pielou's evenness and Simpson's Dominance), the relative benthic index (RBI) and the abundance-biomass comparison method (ABC). Water-sediment interface toxicity bioassays using sea urchin embryos and sediment metal analysis served to check the classifications obtained. The classical community descriptors pointed to a progressive variation in benthic communities along the metal contamination gradient of Portman, but, they did not correctly characterize the environmental status of the lagoon stations. Although the RBI was the index that best classified the sites according to their degree of pollution, the selection of indicator species can bias the results. Since the AMBI, the BENTIX and the ABC method are based on the pollution resulting from organic enrichment, their application in the case of purely toxic pollution may not be successful, as was found to be the case in these two mining-polluted ecosystems. Therefore, the development of new indicator lists according to the type of pollutant may serve to improve the results obtained with organic enrichment-based indices when studying other kinds of disturbance. Finally, we found the toxicity tests to be useful tools for the environmental assessment of aquatic ecosystems, and recommend their inclusion in the WFD. PMID- 15823300 TI - Depositional history of organic contaminants in Narragansett Bay, Rhode Island, USA. AB - Sediment cores were taken at three locations in Narragansett Bay, Rhode Island, USA in 1997 and analyzed for a variety of organic contaminants including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlordanes, linear alkyl benzenes (LABs), benzotriazoles (BZTs) and dichlorodiphenyltrichloroethanes (DDTs) and metabolites. The distributions of these chemicals at Apponaug Cove and in the Seekonk River indicate that there was a disturbance in the depositional environment relative to cores collected at these locations in 1986 demonstrating the potential for buried contaminants to be remobilized in the environment even after a period of burial. Sharp breaks in the concentration of several organic markers with known dates of introduction were successfully used to determine the sedimentation rate at Quonset Point. Both the Quonset Point and Seekonk River cores had subsurface maximums for DDTs, PCBs, PAHs and BZTs, which are consistent with expected inputs to the environment. The Apponaug Cove core showed an increase of most contaminants at the surface indicating a recent event in which more contaminated sediments were deposited at that location. PMID- 15823301 TI - Biodiversity of rocky intertidal benthic communities associated with copper mine tailing discharges in northern Chile. AB - Copper mine tailings have been discharged around the city of Chanaral, in northern Chile, for more than 60 years. This report summarizes a 17-month long monitoring study of species richness and biodiversity at five intertidal sites around the point of the tailing discharge. Total dissolved copper in sites close to the point of discharge varied between 8.72 microg/l and 34.15 microg/l, showing that there has not been a significant reduction since 1994. However, species richness has increased, suggesting a possible recovery of the system. While diversity of sessile organisms correlates negatively with dissolved copper, diversity of mobile invertebrates did not correlate with the metal concentration. To explain the observed results we discuss the role of algal turf interference on the distribution of mobile invertebrates at reference sites, a top-down effect caused by the absence of carnivores at impacted sites, and an avoidance strategy by some species to reduce their contact with contaminated seawater. PMID- 15823302 TI - Inter-population variability in the reproductive morphology of the shore crab (Carcinus maenas): evidence of endocrine disruption in a marine crustacean? AB - Environmental contaminants that are capable of causing endocrine disrupting effects are currently a major cause for concern. These chemicals are known to influence the reproductive development of vertebrates by mimicking or antagonising the actions of endogenous hormones. However, little is known regarding their potential effects on invertebrates. Here we examine variations in the reproductive morphology of the shore crab (Carcinus maenas) for evidence of endocrine disruption. Crabs were collected from a number of sites comprising a putative gradient of exposure to endocrine disrupting chemicals. Patterns of inter-population variability in the expression of sexually dimorphic traits were then examined for evidence of hormone disruption. Extensive variability was detected and patterns of chelal morphology were consistent with the gradient of endocrine disruption. However, overall, the patterns of morphological variability were not consistent with hormonally-mediated effects. This suggests that shore crabs are not susceptible to the same type of endocrine disrupting effects that have been detected in vertebrates, which are most commonly mediated via the oestrogen receptor. However, the potential for androgenic effects on crustacean morphology are discussed. PMID- 15823303 TI - Quantification of in situ nutrient and heavy metal remediation by a small pearl oyster (Pinctada imbricata) farm at Port Stephens, Australia. AB - The use of pearl oysters has recently been proposed as an environmental remediation tool in coastal ecosystems. This study quantified the nitrogen, phosphorus and heavy metal content of the tissue and shell of pearl oysters harvested from a small pearl oyster farm at Port Stephens, Australia. Each tonne of pearl oyster material harvested resulted in approximately 703 g metals, 7452 g nitrogen, and 545 g phosphorus being removed from the waters of Port Stephens. Increasing current farm production of 9.8 tyr(-1) to 499 tyr(-1) would balance current nitrogen loads entering Port Stephens from a small Sewage Treatment Plant (STP) located on its southern shores. Furthermore, manipulation of harvest dates to coincide with oyster condition would likely remove substantially greater quantities of nutrients. This study demonstrates that pearl aquaculture may be used to assist in the removal of pollutants from coastal waters while producing a commercially profitable commodity. PMID- 15823304 TI - The Elbe flood 2002--toxic effects of transported contaminants in flatfish and mussels of the Wadden Sea. AB - Cellular changes in livers from flounder (Platichthys flesus L.) and digestive glands of blue mussels (Mytilus edulis) were studied to analyze whether the Elbe flood catastrophe in August 2002 had any deleterious effects on animal health. Flatfish and mussels were sampled at different locations in the Elbe estuary and the Wadden Sea after the flood wave had reached the German Bight. Internationally used core biomarkers (lysosomal membrane stability, biotransformation enzymes) were applied to assess the toxic effects of putatively increased pollution levels. In comparison to earlier data from long-term studies at the same stations, we found a significant impairment in the function of cell organelles (lysosomes) involved in the detoxification and elimination of pollutants in fish liver. Concentrations of relevant contaminants (organochlorines, polychlorinated biphenyls) were analyzed in parallel with cellular biomarkers, and conspicuously raised concentrations of insecticides metabolites were detected. Cell recovery and a clear reduction in contaminant concentrations were observed in fish livers five months after the flood at all sampling sites except the Helgolander Tiefe Rinne. PMID- 15823305 TI - Revision of the EU Bathing Water Directive: economic costs and benefits. AB - The European Union (EU) Bathing Water Directive of 1976 ([Commission of the European Communities, 1976. Council Directive of 8th December 1975 Concerning the Quality of Bathing Water (76/160/EEC). Official Journal of the European Community. 5th February 1976, L31/1, Brussels]) sets out standards for designated bathing waters which should be complied with by all member states. Intervening advances in pollution science, related technology and managerial expertise have allowed the European Commission to consider revision of EU environmental legislation where appropriate. As a result, a number of revisions to the 1976 Directive have been proposed ([Commission of the European Communities, 1994. Commission Proposal for a Council Directive Concerning the Quality of Bathing Water. Communication from the Commission to the European Parliament and the Council, COM (94) 36 Final, Brussels; Commission of the European Communities, 2000. Developing a New Bathing Water Policy, Communication from the Commission to the European Parliament and the Council, COM (2000) 860 Final, 21/12/200, Brussels; Commission of the European Communities, 2002. Commission Proposal for a Directive of the European Parliament and of the Council Concerning the Quality of Bathing Water, COM (2002) 581 Final, 24/10/2002, Brussels]). This paper considers these revisions in terms of the economic costs and benefits associated with any change in policy. The focus is on the public's willingness to pay for a revised Directive and the consequent public health benefits afforded to individuals and society. These economic benefits are compared to the costs of implementing changes to bring bathing waters up to the required standard. PMID- 15823306 TI - Fate of DDT-related compounds in Bohai Bay and its adjacent Haihe Basin, North China. AB - Concentrations of ten DDTs (2,2-bis-(chlorophenyl)-1,1,1-trichloroethane) of which p,p'-DDA (2,2-bis(chlorophenyl)acetic acid), p,p'-DDM (bis(chlorophenyl)methane) and p,p'-DBP (dichlorobenzophenone) are often neglected, were measured in 25 water and 25 sedimentary samples from Bohai Bay and its adjacent Haihe Basin. The ratio of o,p'-DDX/p,p'-DDX in the upper reaches of Yongdingxin River ranged from 0.71 to 2.44, suggesting that the potential source of pollution would be the manufacturing or use of dicofol near this river. While DDA accounted for 52-93% of the SigmaDDT concentration in water, DDA was only detected in three sedimentary samples. And DDM was found to be an important degradation product of DDT in water following DDA. It should be noted that DBP (0.60-3.30 ng/g) is a major metabolite comparable with DDE (2,2-bis(chlorophenyl) 1,1-dichloroethylene, nondetectable-1.80 ng/g) and DDD (2,2-bis-(chlorophenyl) 1,1-dichloroethane, nondetectable-2.86 ng/g) in sediment in Bohai Bay. Finally, the SigmaDDT concentration in sedimentary sample from Bohai Bay was found to be dependent on the TOC (total organic carbon) value. PMID- 15823307 TI - Ecotoxicity assessment of natural attenuation effects at a historical dumping site in the western Baltic Sea. AB - During the late 1950s and early 1960s of the past century, industrial waste material highly enriched in various contaminants (heavy metals, PAHs) was dumped in the inner Mecklenburg Bay, western Baltic Sea. Large-scale shifts in the spatial distribution of heavy metals in surface sediments were mapped by geochemical monitoring in the mid-1980s and 12 years later in 1997. A further study in 2001 was designed to investigate the small-scale spatial distribution of contaminants inside, on top of, and around the historical dumping ground and to examine possible effects to benthic organisms (Arctica islandica, microbiological toxicity tests). The site is located within an area characterized by a discontinuous deposition of fine sediments, but net sedimentation rate of about 3mm/y led ultimately to a gradual coverage of the dumped material. During the first decades after the dumping, about half of the originally dumped material was already spread over the surrounding area as a result of sediment dynamics and re suspension processes. Recent sediments of the historical dump site are still significantly enriched in heavy metals and PAHs. Microbiological toxicity test results were positive, though uptake of contaminants in mussels was found to deviate only slightly from that of a non-contaminated reference station due to a gradual dilution and decrease in bioavailability of the contaminants. PMID- 15823308 TI - Range extension of two invasive crab species in eastern Australia: Carcinus maenas (Linnaeus) and Pyromaia tuberculata (Lockington). PMID- 15823309 TI - Gender and size effects of metal bioaccumulation on the rock crab, Thalamita crenata, in Dapeng Bay, southwestern Taiwan. PMID- 15823310 TI - Sequential negotiation in multiagent systems for oil spill response decision making. PMID- 15823311 TI - Modeling of depuration of paralytic shellfish toxins in Chlamys nobilis and Perna viridis. PMID- 15823312 TI - Temporal trends in soft tissue metal levels in the periwinkle Littorina littorea along the Scheldt estuary, The Netherlands. PMID- 15823313 TI - Implementing the European Water Framework Directive: the debate continues... PMID- 15823314 TI - A new algorithm for detecting fetal heart rate using ultrasound Doppler signals. AB - Although the fetal heart rate monitoring using ultrasound is widely used it is still not optimized for automatic measurements due to the complexity of the Doppler signal. This paper presents a new fetal heart rate (FHR) detecting algorithm, using sampling auto-correlation approach. The results obtained using the custom-built ultrasonic Doppler fetal heart rate monitoring system are presented and confirm the validity of the method. PMID- 15823315 TI - Ultrasound speckle and equivalent scatterers. AB - B-mode ultrasound images are characterised by the speckle artefact, which introduces fine-false structures whose apparent resolution is beyond the imaging system capabilities. Speckle presence is due to interference effects between overlapping echoes and its occurrence is related to a great number of randomly distributed structure scatterers within a resolution cell. Basing our analysis on linear system theory, we show that a dense random set of scatterers can be substituted by an equivalent one with a much smaller number of periodic scatterers. This new structure with regularly distributed scatterers is able to give rise to the same B-mode image and the same speckle pattern, for a given ultrasound pulse. This new approach helps the understanding of the deterministic nature of speckle and may reduce drastically the computing time in numerical simulations. Additionally, it can contribute to periodicity analysis used in tissue characterisation. PMID- 15823316 TI - Extensions of the scattering-object function and the pulser-receiver impulse response in the field II formalism. AB - The pulse-echo impulse-response format in the Field II formalism is generalized to separately located transmitter and receiver. To first order in sound velocity and density perturbations, identical results for the scattering-object function are obtained for the Morse-Ingard and the Chernov formulation in both the temporal and frequency domains: f(s)=-[2Delta(c/c)+(Delta(rho/rho))(1 cos(theta))] where for ultrasonic pulse-echo or transmission modality, cos(theta) approximately -1 or +1, respectively. PMID- 15823317 TI - Effects of uneven moisture distribution on the strength of and wave velocity in concrete. AB - Earlier findings showed that the effects of moisture (liquid or free water) in hardened concrete on its behavior, especially the lesser known effects ofuneven moisture distribution, can (a) be significant, and (b) vary from property to property. This distribution, for instance whether or not the surface layer is drier than the overall average moisture content, can be characterized by the difference between the velocity of the longitudinal wave (pulse velocity) measured in the standard through-thickness manner, and the velocity of the longitudinal wave propagating on the concrete surface. The summary of earlier findings on the effects of moisture distribution is followed by a recent investigation on pulse velocity in the special case, occurring frequently in practice, when the distribution is uneven because the liquid is concentrated in cracks in the concrete. PMID- 15823318 TI - Dynamic acoustic radiation force acting on cylindrical shells: theory and simulations. AB - An object placed in an acoustic field is known to experience a force due to the transfer of momentum from the wave to the object itself. This force is known to be steady when the incident field is considered to be continuous with constant amplitude. One may define the dynamic (oscillatory) radiation force for a continuous wave-field whose intensity varies slowly with time. This paper extends the theory of the dynamic acoustic radiation force resulting from an amplitude modulated progressive plane wave-field incident on solid cylinders to the case of solid cylindrical shells with particular emphasis on their thickness and contents of their hollow regions. A new factor corresponding to the dynamic radiation force is defined as Y(d) and stands for the dynamic radiation force per unit energy density and unit cross sectional surface. The results of numerical calculations are presented, indicating the ways in which the form of the dynamic radiation force function curves are affected by variations in the material mechanical parameters and by changes in the interior fluid inside the shell's hollow region. It was shown that the dynamic radiation force function Y(d) deviates from the static radiation force function for progressive waves Y(p) when the modulation frequency increases. These results indicate that the theory presented here is broader than the existing theory on cylinders. PMID- 15823319 TI - Coupling an ultrasonic propagation code with a model of the heterogeneity of multipass welds to simulate ultrasonic testing. AB - The knowledge of the anisotropic and heterogeneous behaviour of the material is the key point for understanding ultrasonic testing. In the case of austenitic stainless steel multipass welds, there was no exact knowledge. The succession of passes and complex solidification processes makes the modelling of the resulting grain orientation difficult. The MINA model allows such a description. Coupling this model with the new propagation code ATHENA permits a very good understanding of the propagation of elastic waves in this kind of complex media. After discussing the existing models of grains orientations for multipass welds, we show how the new MINA model allows to simulate the anisotropic heterogeneous structure in a more realistic manner. We introduce the simulated grains structure and the real one, obtained by macrographs, into the ATHENA propagation code. We obtain a very good correlation between the two simulated propagations. A first experimental application is also successful. We show in a quantitative and qualitative way that the modelling results are close to the experimental measurements on a thick austenitic weld. PMID- 15823320 TI - Three-dimensional modelling of micromachined-ultrasonic-transducer arrays operating in water. AB - We report on the 3-D modelling of periodic arrays of capacitive micromachined ultrasonic transducers (cMUTs) operating in fluid. Specific developments have been performed to model biperiodic transducer arrays and to take into account radiation into any stratified media at the front-side as well as the back-side of the device. The model is based on a periodic finite-element-analysis/boundary element-method (FEA/BEM). It is applied to micromachined ultrasonic transducers (MUTs), based on silicon-nitride-circular-membrane arrays on a silicon substrate, and operating in water. The spectrum characteristics of MUTs excited in phase are investigated, showing that very-large-band emission is achievable as previously demonstrated by many authors. However, other contributions are also found, depending on the excitation conditions, that do not radiate in the fluid. These contributions are identified as guided modes that could generate significant cross-talk effects. The origin and the nature of these modes is analyzed to gain insight in the actual operation of MUTs. PMID- 15823321 TI - Directing ultrasound at the cemento-enamel junction (CEJ) of human teeth: I. Asymmetry of ultrasonic path lengths. AB - The diagnosis of degenerative changes in human teeth is of general interest because early detections can avoid greater health problems and further weakening effects. Since the wear of teeth determines their stability and lifetime in relation to the physiological load, an ultrasonic survey of any dimensional changes of the enamel layer and especially of the dentin wall thickness may be very helpful. However, an ultrasonographic diagnosis requires first to determine the anisotropic human tooth properties at clinically relevant locations and to simulate wave propagation phenomena in inhomogeneous tooth models with proper dimensions. The first article of a series that provides modular data of mineralized tissues in human teeth at the cemento-enamel junction (CEJ) deals with an ultrasonic method for measuring the asymmetry of dimensional characteristics of extracted human teeth and their ultrasonic path lengths (UPL). Heavily attenuating tooth halves were investigated with respect to the symmetry of normal and inclined oppositely directed radial ultrasonic paths. The measured UPLs ranged from 1.2 mm to 4.4 mm. The relative difference in inclined UPLs between the left and the right tooth halves reaches almost 30%. This reveals a large asymmetry. The mean difference of angles that represent fastest path lengths was 2.2+/-8.1 degrees, which indicates large asymmetry and anisotropy. Several aspects, which are required for a proper integration of asymmetric data into models designed for medical element engineering and simulation (MEES), are discussed. PMID- 15823322 TI - Design of interdigital transducers for crack detection in plates. AB - Interdigital transducers (IDT) for non-destructive evaluation (NDE) of cracks in plates are designed based on an analytical model established previously. Key considerations include mode selectivity, excitation strength, collimation of wave and cost. The advantage of mode selectivity of IDT over PZT patch is presented both analytically and experimentally. Effects of parameters, namely finger spacing, width, length, number of fingers, and the size of IDT, on the excitation strength and mode selectivity are considered. This led to the design of a mobile double-sided IDT as an efficient device where excitation strength is strong and focused. The device was fabricated in-house using commercially available piezoelectric ceramics and used to develop a procedure for accurate identification of the direction and extent of cracks in plates. Three aluminum plates, one with a linear deep crack, another with a piecewise linear shallower crack and the third with a curved crack, were used to illustrate the accuracy and efficiency of both the proposed device and procedure for effective NDE. PMID- 15823323 TI - Evaluation of interface wave velocity, reflection coefficients and interfacial stiffnesses of contacting surfaces. AB - The phase velocity of the antisymmetric-mode interface wave as well as the longitudinal and shear wave reflection coefficients have been measured for contacting poly(methyl methacrylate) (PMMA) surfaces subjected to different contact pressures. It has been found that while the reflection coefficients decrease as the contact pressure is increased, the phase velocity of the interface wave increases from that of the Rayleigh wave toward that of the bulk shear wave. From these measurements, the normal and tangential interfacial stiffnesses of the contacting PMMA surfaces have been evaluated as functions of the contact pressure. As a result, the two independent procedures to evaluate the tangential stiffness, namely, from the interface wave velocity and from the shear wave reflection measurements, have yielded mutually consistent results. Furthermore, it has been found that the tangential/normal stiffness ratio and the shear/longitudinal reflection ratio of the contact interface are consistent with the predictions of an existing theoretical model for kissing bond interfaces. PMID- 15823324 TI - Biodegradation of phenol and phenol-related compounds by psychrophilic and cold tolerant alpine yeasts. AB - We characterized 32 cold-adapted, psychrophilic and cold-tolerant, yeast strains isolated from alpine habitats with regard to their taxonomy, growth temperature profile, and ability to degrade phenol and 18 phenol-related mono-aromatic compounds at 10 degrees C. Twenty of the strains were identified by sequencing of the ribosomal ITS region as seven species of the basidiomycota: Cryptococcus terreus (three strains), Cryptococcus terricola (one strain), Rhodosporidium lusitaniae (two strains), Rhodotorula creatinivora (10 strains), Rhodotorula ingeniosa (one strain), Mastigobasidium intermedium (one strain), and Sporobolomyces roseus (two strains). Twelve strains sharing closely related ITS sequences could not be identified to the species level; according to their ITS sequence they are included in the Microbotryomycetidae. These 12 strains were psychrophilic (no growth at temperatures above 20 degrees C); one-third of these strains did not grow above 15 degrees C. None of the 32 strains utilized any of the highly volatile mono-aromatic compounds (benzene, toluene, ethylbenzene, nitrobenzene, o-xylene, m-xylene, and p-xylene) as the sole carbon source. Non/low volatile aromatic compounds were degraded in the following order: phenol>hydroquinone>resorcinol>benzoate>catechol>salicylate>>p-cresol>m-cresol. o Cresol, guaiacol, p-nitrophenol, or p-nitrotoluene were not utilized for growth. R. creatinivora strains degraded up to seven compounds, whereas C. terricola and S. roseus strains degraded only two compounds. The toxicity of the compounds was determined via growth inhibition in the presence of toxicants and nutrients at 10 degrees C. R. creatinivora strains were characterized by higher IC50 values than other species, S. roseus was the most sensitive species. The most toxic compounds were the xylene isomers, ethylbenzene, p-nitrophenol, and m-cresol. There was a relation between the chemical structure of the compounds and their toxicity, whereas a relation between the toxicity of the compounds and the ability of the yeasts strains to utilize these compounds for growth was only detected in some cases. PMID- 15823325 TI - Medium composition affects the degree and pattern of cadmium inhibition of naphthalene biodegradation. AB - Metals have been reported to inhibit organic pollutant biodegradation; however, widely varying degrees and patterns of inhibition have been reported. To investigate the roles of medium composition and metal bioavailability on these different degrees and patterns of inhibition, we assessed the impact of cadmium on naphthalene biodegradation by a newly isolated strain of Comamonas testosteroni in three chemically-defined minimal salts media (MSM): Tris-buffered MSM, PIPES-buffered MSM, and Bushnell-Haas medium. Cadmium (total concentrations of 100 and 500 microM) inhibited biodegradation in each medium. Degrees of inhibition were different in each medium. Cadmium was most inhibitory in PIPES buffered MSM and least inhibitory in Bushnell-Haas. For example, in Bushnell-Haas medium, 100 microM cadmium reduced the cell yield more than 4-fold compared to controls not containing cadmium. The same concentration of cadmium completely inhibited growth in PIPES-buffered MSM. No difference in inhibition was observed in any medium when cadmium was added 24 h before inoculation rather than when added within one minute of inoculation. Two patterns of inhibition were observed. Inhibition occurred in a dose dependent pattern in Tris- and PIPES-buffered MSM and in a non-dose dependent pattern in Bushnell-Haas. Specifically, in Bushnell Haas, 100 microM total cadmium extended the lag phase by 23+/-8.66 h, whereas 500 microM did not extend the lag phase. Soluble, ionic cadmium (Cd2+) concentrations were measured and modeled in each medium to assess cadmium bioavailability. In media containing 500 microM total cadmium, bioavailability was highest in Tris- and PIPES-buffered MSM and lowest in Bushnell-Haas. In Bushnell-Haas, cadmium bioavailability was initially higher in the 500 microM treatments (196+/-21.2 microM) than in the 100 microM treatments (78.2+/-2.04 microM); however, after 12 h, bioavailability was higher in the 100 microM treatments (56.4+/-24.8 micro) than the 500 microM treatments (13.3+/-1.2 microM). These data suggest that the type of medium determines the degrees and patterns by which metals inhibit biodegradation and emphasize the importance of coupling metal toxicity and bioavailability data. PMID- 15823326 TI - The effects of wind speed on the relative relationships between different sized fractions of airborne particles. AB - The concentrations of three different size fractions of particulate matter (PM) including PM2.5, PM10, and TSP were determined continuously at hourly intervals from four different sites in Seoul, Korea during the spring of 2001. To learn the effects of wind speed change on PM fractionation, the entire data sets were initially sorted into three particle fractions such as: fine (F: PM2.5), coarse (C: PM10-PM2.5), and giant (G: TSP-PM10). The inter-fraction relationships of PM were then explored by linear regression analysis of the data divided into four wind speed regimes. The results of this analysis, when examined in terms of either relative dominance between different PM fractions (i.e., in terms of their slope values) or strength of correlations, indicate the existence of diverse inter-fraction patterns. Most importantly, the physical influence of wind speed is seen to be reflected most efficiently between fine and coarse particle fractions, as the relative contribution of coarse fraction to the mass concentration of total particles (e.g., PM10) changes proportionally with changes in wind speed. However, such systematic patterns decrease noticeably between fine and giant fractions, as they can be affected more sensitively by such factors as the nature of their sources or the surrounding environmental conditions. The results of our comparative analysis thus confirm that wind speed is a useful barometer to distinguish and predict the behavior of different particle fractions in relation to each other. PMID- 15823327 TI - Comparison of a rhizosphere-based method with other one-step extraction methods for assessing the bioavailability of soil metals to wheat. AB - There is no method recognized as a universal approach for evaluation of bioavailability of heavy metals in soil. Based on the simulation of the rhizosphere soil conditions and integration of the combined effects of root-soil interactions as a whole, a rhizosphere-based method has been proposed. Wet fresh rhizosphere soil was extracted by low-molecular-weight organic acids (LMWOAs) to fractionate metal fractions of soil pools, which were then correlated with the metal contents of wheat roots and shoots. The rhizosphere-based method was compared with other one-step extraction methods using DTPA, EDTA, CaCl2, and NaNO3 as extractants and the first step of the Community Bureau of Reference (BCR) method. Simple correlation and stepwise multiple regression analysis were used for the comparison. Simple correlation indicated that the extractable Cu, Zn, Cr, and Cd of soils by the rhizosphere-based method were significantly correlated with the metal contents of wheat roots. For DTPA, BCR1 and EDTA methods there was a relatively poor correlation between the extractable Cu, Zn and Cd of soil and metal contents of wheat roots. Stepwise multiple regression analysis revealed that the equation of the rhizosphere-based method was the simplest one, and no soil properties variables needed to be added. In contrast, the equations of other one-step extraction methods were more complicated, and soil properties variables needed to be entered. The most distinct feature of the rhizosphere-based method was that the recommended method was suitable for acidic, neutral and near alkaline soils. However, the DTPA and EDTA extraction methods were suitable for calcareous soils only-or-only for acidic soils. The CaCl2, and NaNO3 extraction methods were only suitable for exchangeable metals. In short, the rhizosphere-based method was the most robust approach for evaluation of bioavailability of heavy metals in soils to wheat. PMID- 15823328 TI - Transport of veterinary antibiotics in overland flow following the application of slurry to arable land. AB - The environment may be exposed to veterinary medicines administered to livestock due to the application of organic fertilisers to land. Slurry is often spread on to fields following the harvest of the previous crop. Despite recommendations to do so, the slurry may not be ploughed into the soil for some time. If precipitation occurs before incorporation then it is likely that the slurry and any antibiotic residues in the slurry will be transported towards surface waters in overland flow. This phenomenon has been investigated in a plot study and transport via 'tramlines' has been compared to that through crop stubble. Three veterinary antibiotics, from the tetracycline, sulphonamide and macrolide groups, were applied to the plots in pig slurry. Twenty four hours after the application the plots were irrigated. Following this the plots received natural rainfall. Sulphachloropyridazine was detected in runoff from the tramline plot at a peak concentration of 703.2 microgl(-1) and oxytetracycline at 71.7 microgl(-1). Peak concentrations from the plot that did not contain a tramline were lower at 415.5 and 32 microgl(-1), respectively. In contrast, tylosin was not detected at all. Mass losses of the compounds were also greater from the tramline plot due to greater runoff generation. These did not exceed 0.42% for sulphachloropyridazine and 0.07% for oxytetracycline however. PMID- 15823329 TI - Bioavailability of butachlor and myclobutanil residues in soil to earthworms. AB - To establish chemical extraction procedures for predicting bioavailability of butachlor and myclobutanil in soil, several solvent systems, including methanol, methanol-water (9:1), methanol-water (1:1), acetone-water (5:3), petroleum ether and water, were assessed for their feasibility in determining extractability of the target compounds from soil samples. Experimental data showed that the extractability of butachlor and myclobutanil by the solvents was well linearly correlated with their bioavailability to Eisenia foetida and Allolobophora caliginosa, indicating that these extraction procedures may be efficient for predicting bioavailability of the two pesticides. The concentrations of the pesticides accumulated in E. foetida and A. caliginosa varied with species, suggesting that the availability of the soil-sequestered pesticide is a species dependent process. PMID- 15823330 TI - Photostability of methidathion in wet soil amended with biosolid and a surfactant under solar irradiation. AB - Photodegradation studies of the organophosphorous insecticide methidathion in thin layers of wet soil samples have been carried out under solar irradiation. Soil samples consisted of an agricultural soil added with two amendments: a municipal biosolid and the cationic surfactant TDTMA (tetradecyl trimethyl ammonium bromide). Dark controls of the different soil treatments were also considered. Soil and biosolid samples were previously autoclaved to eliminate biotic degradation. In this study we investigated the role of these amendments in methidathion photodegradation which is a rapid (<7 days) and indirect process. Although scarce differences were found between non-amended and amended samples, methidathion from soil exposed under sunlight is degraded more quickly than in dark conditions. Photodegradation products (methidathion oxon GS 13007 and GS 12956) were not detected. PMID- 15823331 TI - Diel variations in photoinduced oxidation of Hg0 in freshwater. AB - Experiments have been conducted to determine diel variations in photoinduced Hg0 oxidation in lake water under natural Hg0(aq) concentrations. Pseudo-first-order rates of photooxidation (k') were calculated for water freshly collected in a Canadian Shield lake, Lake Croche (45 degrees 56' N, 74 degrees 00' W), at different periods of the day and subsequently incubated in the dark. Hg0 oxidation rates ranged from 0.02 to 0.07 h(-1), increasing from sunrise to noon and then decreasing throughout the remainder of the day. These changes paralleled those in sunlight intensity integrated over 1 h preceding water collection, and suggested that the water freshly collected in daylight was rich in photochemically produced Hg0 oxidants. It was also estimated that under intense solar radiation, even if oxidation rates reached a peak, reduction of Hg(II) was the prevalent redox process. Inversely, Hg0 oxidation overcame DGM production during the night or at periods of weaker light intensity. Overall, these findings explain the decreases in the DGM pool generally observed overnight. They also support previous reports that, during summer days, volatilization of Hg0 from water represent an important step in the Hg cycle in freshwater systems. PMID- 15823332 TI - Characterization and microbial utilization of dissolved organic carbon in groundwater contaminated with chlorophenols. AB - The aim of this study was to characterize the labile part of dissolved organic carbon (DOC) present in groundwater by identification of natural organic carbon substrates and to assess their microbial utilization during aeration of the groundwater. The studied chlorophenol (CP) contaminated groundwater contained 60 2650 micromoll(-1) of DOC of which up to 98.0% were CPs; 1.7% were low-molecular weight organic acids and 0.2% were dissolved free amino acids. Traces of following natural organic carbon substrates were identified: L-alanine, L isoleucine, L-leucine, L-serine, L-threonine, L-tyrosine, L-valine, L-aspartic, acetic, citric, formic, lactic, malic and oxalic acid. Dissolved oxygen concentration inside the CP-plume was lower (mean 25 micromoll(-1)) than outside of the plume (mean 102 micromoll(-1)). Over a monitoring period of four years the concentrations of CPs, Fe(II) and NH4+ were higher inside than outside of the CP plume. Oxygen availability within the CP-plume limits in situ biological oxidation of CPs, DOC, NH4+ and Fe(II). The microbial enzymatic hydrolysis rates of 4-methylumbelliferyl and 7-amino-4-methylcoumarin-linked substrates varied from 0.01 to 52 micromoll(-1)h(-1) and was slightly higher inside than outside the plume. Microbial uptake rates of 14C-acetate, 14C-glucose and 14C-leucine were on average 28, 4 and 4 pmoll(-1)h(-1) outside and 17, 25 and 8 pmoll(-1)h( 1) inside the plume, respectively. The indigenous microorganisms were shown able of hydrolysis of dissolved organic matter, uptake and utilization of natural organic carbon substrates. Therefore, the labile part of DOC serves as a pool of secondary substrates beside the CP-contaminants in the groundwater and possibly help in sustaining the growth of CP-degrading bacteria. PMID- 15823333 TI - A novel enzyme-linked immunosorbent assay system for the quantitative analysis of Carassius auratus vitellogenin. AB - A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantitatively detect Carassius auratus vitellogenin (VTG) levels. The protein levels in fish plasma are useful aquatic biomarkers of estrogenic compounds. This procedure involved an ELISA using monoclonal antibodies of CVmA2 and CVmA7 against Carassius auratus VTG, and CVmA7 conjugated to horseradish peroxidase as the detection antibody. The assay range was between 1 and 401.5 ng/ml and the recovery of the VTG added to Carassius auratus plasma was 92.5-109%. An in vitro assay was performed to measure low levels of the VTG, using primary hepatocytes of Carassius auratus induced by 17-beta estradiol (E2). The detection limit was 1 ng/ml and 137 ng/ml at the maximum. Within each sex of wild Carassius auratus, VTG levels from the river next to sewage treatment works (STWs) were much higher than those from the feeding stream. The Carassius auratus VTG bioassay could be a sensitive and useful tool for quantification of estrogenic principles in aquatic environments. PMID- 15823334 TI - Comparative study of responses in four Datura species to a zinc stress. AB - The effects of zinc toxicity on the growth and the photosynthetic activities of four Datura species (Datura metel, Datura innoxia, Datura sanguinea, Datura tatula) were studied using various ZnSO4 concentrations (0, 1, 2.5 and 5 mM) added in the Coic Lessaint solution. Growth, photosynthesis, chlorophyll fluorescence and chlorophyll concentration were measured after 20 days of zinc stress. These parameters were severely reduced by this heavy metal. The zinc excess involves the stomate closing, the increase of CO2 concentration in the leaves, the inhibition of certain enzyme of the Calvin cycle, a degradation of photosystem and the chlorophyll decomposition. These phenomena allow the decrease of the net photosynthesis to be partially explained. These key parameters to assess photosynthetic performance allow the plants to be classified according to their resistance to zinc. Compared with the three other species, D. innoxia showed a very strong capacity to protect itself against toxic zinc concentrations; a large amount of ZnSO4 (5 mM) was required to inhibit 43% of the photosynthesis. PMID- 15823335 TI - Characterization of a military training site containing 232Thorium. AB - Understanding contaminant distribution is critical to selection and implementation of effective and affordable containment and remediation efforts. This article describes the characterization of soil containing thorium at a training site on Kirtland Air Force Base, Albuquerque, NM. The site has been used by the Defense Nuclear Weapons School since the early 1960's to train personnel in emergency response to nuclear weapons accidents and for characterization and containment of radioactive contamination. The purpose of work reported herein is to describe the primary location and migration pattern of 232Thorium (232Th) and 232Th progeny (decay products) at the site. Soil containing thorium oxide (ThO2) was applied to the site for approximately 30 years (early 1960-1990) and was used to simulate a plutonium release from a nuclear weapons accident. Data presented indicate that surface 232Th and 232Th progeny at approximately 5 times background levels are approaching test site boundaries. However, the data also indicate that vertical migration has not exceeded 0.9 m because of the insoluble nature of ThO2. The major mechanisms of 232Th mobility appear to be surface migration mediated by precipitation runoff and wind-blown soil. PMID- 15823336 TI - Wet deposition of pesticides and nitrophenols at two sites in Denmark: measurements and contributions from regional sources. AB - The concentrations of selected pesticides, of some of their degradation products and nitrophenols in rain were measured at two stations in Denmark in the period January 2000-July 2001. Forty compounds were quantified at least at one station during at least one sampling period. Additionally 17 compounds could be detected but not quantified, and 22 compounds could not be detected at all. The highest depositions of pesticides were observed for pendimethalin and desethylterbuthylazine, which is a degradation product of terbuthylazine. The deposition of the nitrophenols 2,4-dinitrophenol, DNOC, 3-methyl-4-nitrophenol was much higher than that of pesticides. The deposition of 2,4-dinitrophenol was e.g. up to a factor of 40 higher than that of pendimethalin. Atrazine, chloridazon, 2,4-D, dieldrin, disulfoton, fenitrothion, isoproturon, lenacil, metazachlor and propachlor were found in precipitation, although these pesticides are not allowed in Denmark. It can therefore be concluded that they came from abroad and have been transported over at least 60-80 km. For some of these compounds the transport distance is much longer. PMID- 15823337 TI - Molecular orbital studies on brominated diphenyl ethers. Part I--conformational properties. AB - Polybrominated diphenyl ethers (PBDEs) are widely used as additive flame retardants and quantities in the environment are on the rise. Because they are structurally related to polychlorinated biphenyls and also to thyroid hormones, there is serious concern that PBDEs may pose a danger to human health. Knowledge of their conformational properties is key to assessing their environmental fate and risk. The conformational properties of PBDEs were investigated by quantum chemical methods including semiempirical self-consistent field molecular orbital (SCF-MO), ab initio SCF-MO and density functional theory (DFT). Conformational analyses of model congeners 2,2',4,6'-tetrabromodiphenyl ether and 2,3,4,4',5,6 hexabromodiphenyl ether, based on energy maps calculated by semiempirical AM1 method, may indicate that all PBDE congeners except those with the tetra-ortho bromination are conformationally flexible (or soft) due to low energy barriers for interconversion of stable conformers. The results of the conformational analyses are in conformity with recently published X-ray crystallographic data. For comparison with the results of the semiempirical method, higher level ab initio and DFT models were applied as well. The optimized geometries all lie well inside low energy regions on the maps and thus also ascertain the semiempirical calculations. According to computed geometric parameters and net atomic charges, the model B3LYP/3-21G* seemed to give better results than B3LYP/6-31G* and HF/6 31G*. PMID- 15823338 TI - Molecular orbital studies on brominated diphenyl ethers. Part II--reactivity and quantitative structure-activity (property) relationships. AB - Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants and are increasingly turning up in the environment. Their structural similarities to polychlorinated biphenyls and thyroid hormones suggest they may be a risk to human health. The present study examines the reactivity of brominated diphenyl ethers (BDEs) on the basis of the electronic structures as calculated by semiempirical AM1 self-consistent field molecular orbital (SCF-MO) method. Frontier orbital energies were used to elucidate the reactivity of BDEs in electrophilic, nucleophilic and photolytic reactions. From an examination of the frontier electron densities, the regioselectivity, or orientation, of metabolic reactions of BDEs was predicted. Furthermore, satisfactory quantitative structure activity (property) relationship (QSAR and QSPR) models were derived to calculate gas chromatographic and ultraviolet spectral properties and luciferase induction activities from the AM1-computed electronic parameters. PMID- 15823339 TI - A case study of molinate application in a Portuguese rice field: herbicide dissipation and proposal of a clean-up methodology. AB - This study was designed to monitor molinate losses in surface and underground waters during Ordram application in a rice field situated in central Portugal. Water samples were collected from different sites, before, during and about one month and a half after Ordram application. Molinate quantification was based on a solid-phase microextraction (SPME) method followed by gas chromatography with flame photometric detector (GC-FPD) analysis, and led to the conclusion that the herbicide was dissipated in the environment, reaching levels as high as 3.9 microgl(-1) in underground water and 15.8 microgl(-1) in the river receiving tail waters. The feasibility of the application of treatment methodologies based on adsorption or biodegradation as processes to remove molinate from real-world waters was assessed. These methods seem suitable to reduce molinate concentrations to values in the range of the legally recommended limits (<0.5 microgl(-1)). PMID- 15823340 TI - Treatment of chronic endometritis in dairy cows with an intrauterine application of enzymes. A field trial. AB - The use of proteolytic enzymes has been established in the non-antibiotic treatment of mastitis in dairy cattle. The objective of this study was to evaluate, if enzymes are efficacious in the treatment of chronic endometritis. In a controlled field trial, cows with vaginal discharge 21-27 days in milk (DIM) were randomly assigned to two treatment groups. Endometritis was classified into three categories, depending on the type of vaginal discharge: clear mucus with flakes of pus (E1), mucopurulent discharge or fluctuating contents in the uterus (E2), and purulent discharge (E3). In group ENZYMES (n=191), cows received an intrauterine treatment with a salve containing the enzymes trypsin (16 mg), chymotrypsin (16 mg), and papain (8 mg). Cows in group PGF (n=225) were treated with 0.5mg of cloprostenol. Cows that did not show any clinical signs of chronic endometritis were regarded as healthy control group (HC, n=699). In groups ENZYMES and PGF, all cows were re-examined 35-41 DIM. In group ENZYMES, cows were re-treated with enzymes if signs of endometritis were found, while in group PGF all cows received a second dose of cloprostenol, regardless of their clinical findings. Cure rate after the first treatment, defined as the absence of vaginal discharge at the re-examinations, was 59.7 and 68.0% in groups ENZYMES and PGF, respectively (P>0.05). Reproductive performance measures showed no significant differences between the two treatment groups. Service rate was significantly lower for ENZYMES and PGF, respectively, compared to HC. Conception rates to all services and percentages of cows pregnant by 250 DIM were significantly lower in group ENZYMES compared to HC, while no further differences were found between PGF and HC. In both treatment groups, cure rate and reproductive performance measures were better for cows categorized E1 or E2, than for cows categorized E3, respectively. Conception rate to all services for cows with endometritis category E1 was higher in group PGF than in group ENZYMES (P<0.05). The results of this field trial suggest that prostaglandin F(2alpha) is still the treatment of choice for chronic endometritis in dairy cattle. PMID- 15823341 TI - Conception rate after fixed time insemination following ovsynch protocol with and without progesterone supplementation in cyclic and non-cyclic Mediterranean Italian buffaloes (Bubalus bubalis). AB - The aim of this study was to test the effect of progesterone supplementation to Ovsynch protocol in cyclic and non-cyclic Mediterranean Italian buffaloes on conception rate after fixed time artificial insemination. From 169 pluriparous buffaloes, 2 groups were identified and subjected to: (1) Ovsynch protocol (OV; n=83) and (2) Ovsynch protocol with the supplementation of progesterone from days 0 to 7 (OV+PROG.; n=86). All cows were inseminated 16-20 h after the second GnRH administration. Within each group, non-cyclic buffaloes were identified (OV=21 and OV+PROG.=20). Overall conception rate was significantly higher in cyclic compared to non-cyclic buffaloes: 43.7% versus 17.0%, respectively, P=0.001. A significant effect of progesterone supplementation on conception rate was observed in non-cyclic buffaloes (30% versus 4.7%, P=0.04) but not in cyclic buffaloes (51.5% versus 35.7%, P=0.077). Collectively, the presence of a large follicle (>or=10 mm) detected at the beginning of the Ovsynch protocol by ultrasound significantly affected conception rate (44% versus 8%, P=0.01). The findings of the present study suggest that (i) progesterone supplementation to the Ovsynch protocol in buffaloes increases conception rate in non-cyclic animals, (ii) the presence of a large follicle at the beginning of the Ovsynch protocol is a determining factor for a successful synchronization of ovulation and high conception rates and (iii) ultrasound monitoring can improve the overall efficiency by selectively identifying more suitable cycling animals carrying a responsive follicle at the time of first GnRH administration. PMID- 15823342 TI - Integrating ultrasonography within the reproductive management of the collared peccary (Tayassu tajacu). AB - Ultrasound imaging has been used to elucidate certain aspects of the reproductive biology of wild or endangered species. However, to our knowledge, this tool has not been used for reproductive monitoring of the collared peccary (Tayassu tajacu). In this study, real-time ultrasonography was used in 16 collared peccary females to diagnose early pregnancy status and predict gestational age. Based on the detection of an embryo, the earliest pregnancy diagnosis was made on Day 18 after mating, with the mean time needed for diagnosis being 22 days. Overall accuracies on Days 22, 26 and 28 were 56, 93, and 100%, respectively. On Days 26 and 28, all pregnancy and non-pregnancy diagnoses, respectively, were correct. The fetal measurements that best correlated with gestational age were crown-rump length (CRL) and the length and diameter of the thorax. CRL was considered the most practical measurement because, contrary to thoracic fetometry, it could be determined when the embryo was first detected. Our findings revealed real-time ultrasound scanning to be a very accurate method for early pregnancy diagnosis and prediction of gestational age in the collared peccary. PMID- 15823343 TI - Progestagen profiles during the last trimester of gestation in Thoroughbred mares with normal or compromised pregnancies. AB - Progesterone (P4), pregnenolone (P5) and their metabolites are present in maternal plasma in pregnant mares. It is believed that one of these progestagens may maintain myometrial quiescence. The aims of this study were to identify specific progestagens in pregnant mares' plasma and determine whether these differed between mares with healthy or compromised pregnancies. Jugular blood samples were collected between 243 and 351 days gestation from 19 healthy Thoroughbred mares and 14 mares with placental pathology, including placentitis, and other clinical problems (uterine torsion/rupture, colic, laminitis). Ten progestagens were identified using gas chromatography-mass spectrometry, of which seven increased significantly with gestational age in healthy mares while P4 was undetectable. Mares with placentitis had increased concentrations of either P5 and/or P4 and several metabolites (5alpha-DHP, P5betabeta, betabeta-diol, betaalpha-diol, 20alpha-5P) suggesting increased fetal production of P5 and/or P4 and increased metabolism in the utero-placental tissues in response to chronic stress. Mares with other placental pathology had raised P4 concentrations while 5alpha-DHP and 3beta-5P were low possibly due to reduced placental function. In mares with problems unrelated to the placenta, most progestagens were substantially lower than control values. Although progestagen profiles differed between normal and abnormal pregnancies, no clear link was demonstrated between maternal plasma concentrations of P4, 5alpha-DHP or any other progestagen and the maintenance of pregnancy. PMID- 15823344 TI - Season effect on genitalia and epididymal sperm from Iberian red deer, roe deer and Cantabrian chamois. AB - Seasonality deeply affects the physiology and behavior of many species, and must be taken into account when biological resource banks (BRBs) are established. We have studied the effect of seasonality on many reproductive parameters of free ranging Iberian red deer, roe deer and Cantabrian chamois, living in Spain. Testicles from hunted animals were collected and sent to our laboratory at different times during the year. We recorded the weight and volume of testis, the weight of the epididymis and its separate parts (caput, corpus, and cauda), the weight of the sperm sample collected from the cauda epididymis, and several sperm parameters (sperm concentration, spermatozoa recovered, motility, HOS test reactivity, acrosomal status, and viability). We studied the data according to several periods, defined accordingly to each species. For red deer, we defined rut (mid-September to mid-October), post-rut (mid-October to mid-December), and non-breeding season (February). For roe deer, they were pre-rut (June), rut (July), post-rut (first fortnight of August), and non-breeding season (September). For chamois: non-breeding season (June to mid-September) and breeding season (October-November). The rut/breeding season yielded significantly higher numbers for almost all parameters. However, in the case of red deer, sperm quality was higher in the post-rut. For roe deer, testicular weight was similar in the pre-rut and in the rut, and sperm quality did not differ significantly between these two periods, although we noticed higher values in the rut. In the case of chamois, sperm quality did not differ significantly from the breeding season, but data distribution suggested that in the non-breeding season there are less males with sperm of good quality. On the whole, we find these results of interest for BRB planning. The best season to collect sperm in this species would be the breeding season. However, post-rut in red deer, pre-rut in roe deer, and non-breeding season in chamois could be used too, because of the acceptable sperm quality, despite the lower quantity salvaged. More in-depth research needs to be carried out on the quality of sperm salvaged at different times of the year in order to confirm these findings. PMID- 15823345 TI - Enhanced efficiency in the production of offspring from 4- to 8-week-old lambs. AB - The rate of production of offspring from lambs following the transfer of embryos produced by in vitro maturation and fertilisation has been disappointing. This study was conducted on the rationale that gonadotropin treatment protocols used in the past failed to maximise oocyte quality. Consequently, three experiments were conducted in Merino lambs to examine: (1) the method of FSH administration (single versus multiple treatments), (2) the time of oocyte collection (48, 60 or 72 h after the first FSH treatment), (3) progesterone treatment and (4) eCG treatment and its timing on oocyte yield, cleavage rate and blastocyst formation rate. No factor significantly influenced oocyte yield, but all significantly influenced the rates of cleavage and/or blastocyst formation in one or more experiments. The preferred protocol consisted of the administration of 4x40 mg of FSH, no progesterone treatment and with 500 IU of eCG given at the time of the last FSH treatment. The optimal time of oocyte collection was 48 h after the first FSH treatment. A field evaluation, involving lambs from five generations, resulted in the production of 9.0-13.9 lambs per donor lamb; this protocol generated more lambs per donor lamb than protocols previously reported. PMID- 15823346 TI - Accessory sperm: a biomonitor of boar sperm fertilization capacity. AB - The number of accessory sperm found in the zona pellucida of porcine embryos was correlated to their individual quality and to the embryo quality range found within a single sow. Our goal was to determine whether accessory sperm counts provide semen evaluation with additional, useful information. Accessory sperm count was highest when only normal embryos were found in a given sow and diminished if oocytes or degenerated embryos were present (P<0.01). Within a given sow, normal embryos had higher (P<0.05) accessory sperm counts than degenerated embryos, although not when oocytes were also present. Fertilization capacity of sperm is optimal when only normal embryos are found in a given sow; this capacity is indicated by high accessory sperm counts. A decrease in fertilization capacity is reflected in diminishing accessory sperm counts. The boar had a significant effect (P<0.01) on accessory sperm count, but not on the percentage of normal embryos; this suggests that accessory sperm may be more sensitive indicators of the fertilization capacity of sperm than the percentage of normal embryos. We conclude that accessory sperm count can be used for the detection of compensable defects in sperm and is a valid parameter for assessing sperm fertilization capacity. PMID- 15823347 TI - Characterisation of low, medium and high responders following FSH stimulation prior to ultrasound-guided transvaginal oocyte retrieval in cows. AB - In human IVF, the concept of 'low responders' is well known and generally defined as women with poor-response to gonadotrophin stimulation in a previous induction cycle. The objective of this retrospective study is to describe and characterise the concepts of 'low-, medium-, and high-response' and 'low, medium, and high responders' in bovine-assisted reproduction by analysing the OPU-IVF results obtained following 665 gonadotrophin-stimulated sessions conducted in 112 animals, nearly all of which were previously unsuccessful in traditional multiple ovulation and embryo transfer (MOET) programs. They were submitted to OPU and IVP between 1999 and 2003. In reference to these 665 OPU sessions, categories of response were defined based on the overall mean+/-S.D. follicles aspirated and COC obtained i.e., for follicles 14.7+/-9.8 and for COCs 11.7+/-8.1. So arbitrary cut-off values to define the categories of sessions were for follicles 5 and 25, and for COC 4 and 20. The three categories for follicles punctured in one session were therefore follicle low-response (FLR)or=25 follicles and for COCs recovered in one session COC low-response (CLR)or=20 COC. In addition, four categories of animals were also defined: (1) a low responder animal (LRA) had at least one OPU session in which FLR and CLR were observed (genuine low-response, see Section ); these animals did not have any high-response sessions, (2) a medium responder animal (MeRA) had only medium-responses, (3) a high responder animal (HRA) had at least one OPU session in which FHR and CHR were observed; these animals did not have any low-response sessions, and (4) mixed responder animals (MiRA) had both low and high-responses. Finally, we distinguished biological (animals) and technical (recovery rate and ultrasound resolution) causes of response differences. In 'low, high, medium and mixed reponders,' different results were obtained (p<0.05): mean follicle numbers (8.8+/-4.8a, 22.4+/-10.5c, 13.2+/-5.2b,15.1+/-10.2d), COC numbers (6.3+/-3.9a, 18.5+/-8.2c, 10.4+/-4b, 12.0+/-8.3d), embryo numbers (1.8+/-2.1a, 5.6+/-4.9c, 2.5+/-2.7b, 3.5+/-3.8d) and also for recovery rate (72%a, 83%b,79%, 79%) and percentage embryo development (29%, 30%a, 24%b, 29%). In conclusion, the results of this study demonstrate that variability in OPU results has technical (ultrasound resolution and recovery rate) as well as biological (animal) aspects. Selection of animals with extreme (high or low) follicle and COC production results allows us to distinguish three populations: 'low, medium, and high responders' to FSH stimulation. PMID- 15823348 TI - Use of a homologous radioimmunoassay (RIA) to evaluate the effect of maternal and foetal parameters on pregnancy-associated glycoprotein (PAG) concentrations in sheep. AB - Early pregnancy detection and prediction of the number of lambs would be profitable for sheep breeders because it enables them to adjust nourishment of pregnant ewes according to the individual needs in order to prevent health problems around parturition. The concentration of ovPAG has previously been reported to be related with maternal parameters (farm, breed and age) as well as foetal parameters (number of lambs, their sex and birth weight), but contradictory results were obtained in different small-scale studies. This large scale study evaluates the effect of these parameters on the ovPAG concentration, determined by a homologous radioimmunoassay (RIA), and it further investigates the possibility to predict the number of lambs by means of homologous ovPAG determination. Eighty-three and ninety-five ewes of the Suffolk and Texel breed, respectively, housed on four different farms (experiment 1) and 68 ewes of the Suffolk breed, housed on two different farms (experiment 2) were included in this study, and their estrous cycles were synchronised using a progesterone analogue. On the day of synchronisation (D-14) and at 25 (D25), 35 (D35) and 45 (D45) days after insemination, blood samples were taken and a homologous radioimmunoassay (RIA) was used to determine the ovPAG concentrations. At parturition, age of the ewe, number and sex of the lambs (experiment 1) and birth weight (experiment 2) were registered. OvPAG concentrations were not affected by age of the ewe and sex of the lambs. Farm and breed of the ewes, number and birth weight of the lambs had a significant effect on ovPAG concentrations at all time points (P<0.05). The odds of multiple lambs increased significantly with increasing ovPAG concentration, although prediction of litter size based on ovPAG concentration at the individual ewe level was not useful due to small sensitivity and/or specificity whatever the cutoff value used. In conclusion, the ovPAG concentration is affected by farm and breed of the ewes, and number and birth weight of the lambs. PMID- 15823349 TI - Bovine embryo development after IVF with spermatozoa having abnormal morphology. AB - The study was conducted to evaluate the effects of scrotal insulation on semen samples collected from bulls on embryonic development after IVF. Semen samples were obtained and cryopreserved from four Holstein bulls before and after a scrotal insulation period of 48 h (Day 0). Three types of samples were used for IVF: (1) semen from the test bulls collected 5 d prior to scrotal insulation (pre insult); (2) semen from Day 13 (2-week post-insult; 2-week PI); and (3) semen from Day 20 (3-week PI). After 18 h of sperm-oocyte co-incubation, the zygotes were cultured for 8 d when a developmental score (0=degenerate, 1=2-cell embryo through 5=blastocyst) was assigned to each embryo. The post-thaw morphological evaluation of sperm samples revealed a decrease (P<0.01) in the percentages of normal spermatozoa in the 3-week PI samples in comparison with the pre-insult samples for Bulls I and III (74-22.3% and 67.7-0.5 %, respectively). The percentage of vacuolated spermatozoa increased significantly for Bull II. The cleavage and blastocyst formation rates and embryo development scores were affected (P<0.01) by the interaction of bull by sample collection time. For Bulls I and III (severe responders) the scrotal insulation effects persisted from the time of cleavage through blastocyst formation. In contrast, the cleavage and blastocyst formation rates for Bulls II and IV were unaffected, despite high percentages of vacuolated spermatozoa present in the post-insult samples for Bull II. In conclusion, the use of scrotal insulation to elevate scrotal temperature was an effective method to obtain semen samples with high percentages of abnormal spermatozoa. The decrease in embryonic development after IVF when using spermatozoa with morphological abnormalities seemed to be multifaceted and related to changes in head morphology. PMID- 15823350 TI - The effect of hCG administration five days after insemination on the first service conception rate of anestrous dairy cows. AB - The aim of this study was to investigate the effect of treating anovulatory anestrous (AA) dairy cows with 1500 IU of hCG IM, 5 d after insemination, on their first service conception rate. A clinical trial was conducted during the 2003/2004 breeding season involving 442 AA dairy cows in six herds. On Day -8, all cows were treated with a progesterone-containing intravaginal device (Cue Mate). The devices were removed on Day -2, and on Day -1 all cows received an IM injection of 1mg of estradiol benzoate. Cows in the control group (n=220) received no further treatments. Cows in the treatment group (n=222) which had been inseminated on Days 0 or 1 were treated with 1500 IU of hCG IM 5 d after insemination. Blood was collected from 30 cows (15 in each group) on Days 5 and 12 after AI for analysis of plasma P4 concentration. There was no difference in first service conception rates between the control and treatment groups (46.3% versus 43.6%, respectively; P=0.68), despite the fact that plasma P4 concentrations were higher in the treatment group on Day 12 (4.9+/-1.3 ng/mL versus 6.2+/-2.7 ng/mL for control and treatment groups, respectively; P<0.01). In conclusion, 1500 IU of hCG 5 d after insemination did not improve first service conception rate in AA dairy cows. PMID- 15823351 TI - Experimental microbial contamination and disinfection of dry (vapour) shipper dewars designed for short-term storage and transportation of cryopreserved germplasm and other biological specimens. AB - Cryopreservation, storage and transport of cryopreserved germplasm without the risk of disease transmission is of great concern to animal and human health authorities. Here we report on the efficacy of microbial decontamination of the liquid nitrogen (LN) dry (vapour) shippers used for short-term storage and transportation of germplasm and other biological specimens. Dry shippers containing either a hydrophobic or a non-hydrophobic LN absorbent were experimentally contaminated with high titers of cultures of Pseudomonas aeruginosa, Escherichia coli, Staphylococus aureus, bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1). Biocidals with broad spectrum antimicrobial activity and gas vapours of formalin and ethylene oxide were used for disinfection of the dewars. Among the biocidals used, treatment with sodium hypochlorite solution, the quaternary ammonium-based disinfectants and peracetic acid were the most effective and useful for dry shippers with a hydrophobic LN absorbent. None of the bacterial or viral microorganisms were detected in samples of semen and embryos stored in dry shippers following their disinfection with these biocides. An application of some other disinfectants, due to their foaming properties or to the permeability of the absorbent hydrophobic membrane (HM) was not effective or may have caused irreversible damage to the LN absorbent. Gas sterilization by ethylene oxide in contrast to formalin was fully effective for both types of dry shippers. PMID- 15823352 TI - Induction of parturition in the bitch with the progesterone-receptor blocker aglepristone. AB - The triggering mechanism for parturition in the bitch remains unclear. Consequently, the development of drugs to successfully induce parturition in the dog has been difficult. The aim of this study was to evaluate the efficacy of the progesterone-receptor blocker aglepristone for the induction of parturition in beagle bitches. The course of parturition was therefore investigated in six parturitions induced by aglepristone and in six spontaneous parturitions. In addition, data were collected on pup survival and growth rates. Aglepristone was administered twice with a 9h interval on day 58 of pregnancy. If parturition did not proceed a standard intervention protocol was applied. Expulsion of the first pup occurred between 32 and 56 h after the first treatment with aglepristone, at which time the plasma progesterone concentration was still elevated. Accordingly, the gestation length of the bitches in the induced group (59.5+/-0.2 days) was significantly shorter than that of the spontaneously whelping bitches (62.2+/-0.5 days). The expulsion phase length, the inter-pup interval, the number of puppies born dead, and the number of clinical interventions needed during parturition did not significantly differ between the spontaneously whelping and the induced group. Pup survival and mean birth weights in the two groups did not differ significantly and aglepristone treatment had no significant influence on the growth rates. The results of this study show that aglepristone is an effective drug which can be used safely for the induction of parturition in the dog. PMID- 15823353 TI - The effects of previous ovarian status on ovulation rate and early embryo development in response to superovulatory FSH treatments in sheep. AB - A total of 64 ewes was used to determine if the changes in superovulatory yields related to the ovarian status at the start of superovulatory treatment are due to differences in the population of gonadotrophin-responsive follicles, alterations in the processes of ovulation or transport of embryos from oviduct to uterus and/or developmental competence of the oocyte/embryo. Ovarian status at the start of a superovulatory FSH step-down treatment, administered coincidentally with a progestagen, was assessed by ultrasonography. On Day 4 after progestagen withdrawal, embryos were recovered from oviduct and their viability was determined by assessing development in vitro culture (IVC) until the hatched blastocyst stage. In all the ewes, the ovulation rate was related positively to the number of 2-3 mm follicles at first FSH injection (P<0.005). However, the total number of embryos and their viability were related to the more limited category of 3 mm follicles (P<0.05), whereas a higher degeneration rate was related to the number of 2mm follicles. The presence of a corpus luteum (CL) at the start of superovulatory treatment exerted a protective effect on embryonic viability, decreasing the degeneration of embryos. On the other hand, the presence of a dominant follicle at first FSH dose affected the mean size of the pool of follicles responding to the superovulation treatment, because ovulation arose from 3 to 5 mm follicles in absence of large follicles (P<0.05), but from 2 to 3 mm follicles when large follicles were present (P<0.005), indicating atresia in medium sized follicles in the presence of a large follicle. PMID- 15823354 TI - Effects of aromatic cationic molecules on bovine viral diarrhea virus and embryonic development. AB - Bovine viral diarrhea virus (BVDV) has been shown to replicate in embryo culture systems and remain associated with bovine embryos developing in vitro. In this study, novel antiviral agents were evaluated for capability to inhibit replication of BVDV without affecting embryonic development. Serial concentrations of 2-[5(6)-{2-imidazolinyl}-2-benzimidazolyl]-5-(4 aminophenyl)furan (DB456) or 2-(4-[2-imidazolinyl]phenyl)-5-(4 methoxyphenyl)furan (DB606) were prepared in IVC medium. Then, bovine uterine tubal epithelial cells (UTC) were placed in IVC media with varying concentrations of DB456 or DB606. Within 1h, a genotype I or II strain of BVDV was added to the cultures. Cultures were maintained for 7 days. Infectious virus was quantitated in IVC media collected on days 3 and 7 and in UTC lysates harvested on day 7. The effective antiviral concentrations of DB606 were much lower than effective antiviral concentrations of DB456. In subsequent experiments, IVF presumptive zygotes were cultured in IVC medium with or without DB456 or DB606 at multiple concentrations for 7 days to evaluate effect of the compound on conceptus development. On day 7, stage of embryonic development was observed, and blastocysts were harvested and stained using Hoechst 33342 to enumerate embryonic cells. While DB456 inhibited blastocyst development, DB606 at 20 times the effective antiviral concentration did not hinder blastocyst development or reduce the mean number of cells per blastocyst. These preliminary results indicated that bovine embryo cultures might be safely supplemented with effective concentrations of an antiviral agent. PMID- 15823355 TI - Differences between Belclare and Suffolk ewes in fertilization rate, embryo quality and accessory sperm number after cervical or laparoscopic artificial insemination. AB - Ewe breed has been shown to have a major effect on pregnancy rates following cervical AI using frozen-thawed semen. The main objective of this study was to examine the differences between purebred Belclare and Suffolk ewes (multiparous) in fertilization rate, number of accessory sperm and stage of embryo development on day 6 after cervical or laparoscopic AI with frozen-thawed semen. In experiment 1, Belclare and Suffolk ewes were synchronized for 12 days and were either cervically inseminated (year 1: n=28 and 31; year 2: n=16 and 15, respectively) or laparoscopically inseminated (year 2: n=13 and 14). In experiment 2, superovulated Belclare (n=4) and Suffolk (n=13) ewes were laparoscopically inseminated. All ewes were slaughtered 6 days after AI; oocytes/embryos were recovered, morphologically graded and stained to assess the number of cells and accessory spermatozoa. Data from both experiments were combined for statistical analysis. The proportion of ewes with fertilized oocytes was significantly higher following laparoscopic AI compared with cervical AI (54% versus 19%). More Belclare than Suffolk ewes yielded fertilized oocyte(s) after cervical AI (34% versus 10%, P<0.02) but there was no difference after laparoscopic AI (62% versus 60%). From the ewes that yielded at least one fertilized oocyte the proportion of Belclare ewes with embryos at the morula/blastocyst stage was significantly greater than for Suffolk ewes (94% versus 59%, P<0.02). A higher proportion of Belclare than Suffolk ewes had evidence of sperm reaching the site of fertilization following cervical AI (39% versus 15%, P<0.02) but there was no difference after laparoscopic AI (62% versus 64%, P>0.8). Amongst the ewes with evidence of sperm at the site of fertilization, laparoscopic AI resulted in a higher number of sperm per oocyte/embryo or per ewe than cervical AI (P<0.01). These results suggested that the difference in pregnancy rate between Suffolk and Belclare ewes following cervical AI was due to: (i) sperm traversing the cervix and uterus in a higher proportion of Belclare than Suffolk ewes, leading to a higher incidence of fertilization and (ii) the lower developmental competence of fertilized oocytes from Suffolk ewes. PMID- 15823356 TI - Increasing storage time of extended boar semen reduces sperm DNA integrity. AB - There is an extensive use of artificial insemination (AI) in the pig industry. Extended liquid boar semen may be used for insemination for up to 5 days after collection. The objective of this study was to determine the changes in sperm quality, when boar semen was extended and stored at 18 degrees C for up to 72 h post-collection. The study included three ejaculates from five boars, for each of the four breeds: Duroc, Hampshire, Landrace and Danish Large White (n=60 ejaculates). The sperm chromatin structure assay (SCSA) showed an increase in DNA fragmentation index (DFI) after 72 h of incubation (P<0.001), with no differences between breeds (P=0.07). For two Hampshire boars, all ejaculates had a large increase in DFI after 24 h of incubation. The standard deviation of DFI (SD-DFI) differed between breeds, with the SD-DFI for Hampshire being significantly greater than for the other breeds. The SD-DFI did not change during the 72 h of storage. Sperm viability was determined using SYBR-14 and propidium iodide in combination with flow cytometry. The sperm viability did not differ between breeds (P=0.21), but a difference in viability during storage (P<0.001) was detected. In conclusion, the SCSA cytogram patterns were consistent for different ejaculates within boars and storage of extended boar semen at 18 degrees C for 72 h significantly decreased the integrity of sperm DNA. PMID- 15823357 TI - The effect of nutrition during pregnancy on the in vitro production of embryos from resulting lambs. AB - It is possible to produce offspring from FSH-treated lambs using in vitro maturation and fertilisation procedures but a major constraint is the high embryo wastage after transfer. It is postulated that this wastage is associated, at least in part, with the quality of the harvested oocytes. The aim of this study was thus to determine if nutrition during pregnancy influenced the quality of oocytes collected from resulting lambs. The study was a 2x2x2 factorial that examined the effect of a low (L; 0.7x maintenance) or high (H; 1.5x maintenance) diet provided during three periods (-82 to 70, 71-100 and 101-126 days relative to the date of conception). There were eight treatments namely LLL, LLH, LHL, LHH, HLL, HLH, HHL and HHH. Oocytes were harvested from 9-week-old lambs, matured and fertilised in vitro and the percentages of oocytes and embryos that developed into blastocysts were recorded. There were significant differences between treatments in oocyte and embryo yields and these resulted from complex interactions between diet and the stage of pregnancy. The efficiency of producing blastocysts from oocytes was highest when a H diet was provided between 71 and 110 and/or 101-126 days of pregnancy. These results demonstrate the need to manage nutrition during pregnancy in programs aimed at producing offspring from juvenile animals. PMID- 15823358 TI - Effect of EGF on in vitro maturation of domestic cat oocytes. AB - The objective of this study was to evaluate the influence of different concentrations of epidermal growth factor (EGF) on in vitro maturation of domestic cat oocytes. A total of 444 cat oocytes were matured in MSOF (maturation synthetic oviductal fluid) in the presence of varying EGF concentrations: (1) MSOF (control); (2) MSOF+10 ng/mL EGF (EGF10); (3) MSOF+25 ng/mL EGF (EGF25); and (4) MSOF+50 ng/mL EGF (EGF50). After IVM, oocytes were in vitro fertilized to verify the effect of adding EGF on cytoplasmic maturation. Cleavage rate was recorded and noncleaving oocytes were stained with Hoechst 33258 and examined to determine nuclear maturation rate. Cleaved zygotes were cultured in vitro and embryo stages were evaluated on days 6 and 7. There was no difference among groups in the total number of oocytes reaching the metaphase II (MII) stage (P>0.05). The EGF25 group had the highest (P<0.01) blastocyst yield (37.5%) and developmental competence (60.9%). Cleavage rate and resulting morulae and blastocysts on day 6 for EGF25 group were higher (P<0.01) than control and EGF50 groups. Although EGF did not significantly enhance nuclear maturation rate, it had a dose-related positive effect on cytoplasmic maturation, since the oocyte's ability to cleave and reach the blastocyst stage was improved at 25 ng/mL, with intermediate improvement at 10 ng/mL, but 50 ng/mL had no significant benefit. In conclusion, the addition of EGF to the maturation medium enhanced cytoplasmic maturation of cat oocytes in vitro. PMID- 15823359 TI - The effect of oxygen tension on porcine embryonic development is dependent on embryo type. AB - Reducing oxygen concentration from atmospheric levels during in vitro culture generally, but not invariably, improves embryonic development across a range of species. Since the few published reports of such an action in the pig are contradictory--perhaps a consequence of the derivation of the embryos prior to culture--a study was performed to examine the effect of O2 tension during culture on three different types of porcine embryos, namely: in vivo flushed embryos, and in vitro matured oocytes either fertilized in vitro or parthenogenetically activated. In vivo embryos (n=208) were flushed at the 2-8 cell stage. Cumulus oocyte complexes (COCs) destined for IVF or parthenogenetic activation were derived from 2 to 6 mm, post-pubertal ovarian follicles and matured for 48 h in TCM-199. Parthenogenones were generated by activating denuded oocytes (n=573) with 10 mM calcium ionophore, followed by 2 mM DMAP prior to culture. The IVF embryos (n=971) were produced by fertilizing COCs (day 0) with fresh ejaculated semen in modified tris-based medium for 6 h before cumulus removal. All embryos were cultured in BECM-3 containing 12 mg/mL fatty-acid-free BSA up to day 4, followed by BECM-3 supplemented with 10% calf serum until day 7. The gas environment for IVM/IVF was 5% CO2 in air, while that for IVC was either 5% CO2 in air or 5% O2, 5% CO2 and 90% N2. Low O2 tension increased both day 7 blastocyst rates (high versus low O2, respectively; 9.3+/-2.9%: 26/280; 23.9+/ 4.2%: 71/293; P<0.001) and total cell numbers (39.3+/-2.9, n=24 versus 61.2+/ 7.7, n=61; P=0.01) of parthenogenetically activated embryos. In contrast, such a treatment neither affected blastocyst rates (89.3+/-6.9 versus 87.8+/-7.5) nor cell numbers (87.4+/-4.5 versus 87.7+/-4.8) of in vivo flushed embryos. The effect of reduced O2 concentration on IVF embryos was intermediate, since only cell numbers were improved (69.8+/-3.5, range=17-204, n=49; 88.5+/-5.8, range=28 216; n=66; P<0.01), equivalent to that recorded in in vivo flushed embryos. However, blastocyst rates were unaffected (10.7+/-1.4%: 51/486; 12.9+/-2.2%: 67/485). The effect, when present, of reducing O2 concentration from 20 to 5% was beneficial for pig in vitro embryonic development. The responses are apparently dependent on firstly, the manner by which the embryonic cell cycle is activated and secondly, the derivation of the tissue prior to placement into culture, if the observed resilience of in vivo embryos is independent of treatment duration. PMID- 15823360 TI - BSP A1/A2-like proteins in ram seminal plasma. AB - The objective of this study was to assess the protein profile of ovine seminal plasma using 2D-PAGE and verify if BSP A1/A2 are present in ovine seminal plasma. Seminal plasma was collected from three mature rams and pooled to eliminate individual differences. Seminal plasma samples were submitted to 2D-PAGE using 12% acrylamide gels. The image analysis software identified 21 protein spots on the air-dried gel, with molecular weight ranging from 15 to 115 kDa and pI 3.2 to 8.7. The most prominent spots were those <30 kDa. The most intensely stained spots were: 3 (18-19 kDa, pI 4.8-5.0), 5 (17-18 kDa, pI 5.0-5.2), 7 (15-16 kDa, pI 6.2-6.4), and 23 (105-108 kDa, pI 6.8-7.0). Three of these spots (spots 3, 5 and 7, respectively) accounted for 41.1% of the relative intensity of the spots of the gels, based on the intensity of the Comassie blue staining. Western blot analysis indicated that spots 3 and 5 were similar to BSP A1/A2 (16.5, pI 4.7-5.0 and 16 kDa, pI 4.9-5.2) identified in Manjunath's studies [Manjunath P, Sairam MR. Purification and biochimical characterization of three major acid proteins (BSP A1, BSP A2 and BSP A3) from bovine seminal plasma. Biochem J 7 (1987) 685 92.], based on the specific reaction of the polyclonal antibody to those spots. PMID- 15823361 TI - The effect of ascorbic acid supplementation on sperm quality, lipid peroxidation and testosterone levels of male Wistar rats. AB - This study was conducted to investigate the effects of ascorbic acid supplementation in drinking water on semen quality, lipid peroxidation and plasma testosterone level of male rats. In this investigation, 24 male Wistar rats were used. The animals were divided into three group, and 500, 250 and 0 (control) mg/kg/day ascorbic acid were supplemented with drinking water of rats in Groups A, B and C during 8 weeks, respectively. Ascorbic acid supplementation did not increase in the body weight and weights of the testis, epididymis, seminal vesicles and ventral prostate. Exogenous supplementation with ascorbic acid significantly increased (P<0.05) the concentration of ascorbic acid in the testes and blood plasma, and the level of lipid peroxidation significantly decreased (P<0.05) in these locations. There was no significant difference in spermatozoon motility among the three groups. However, epididymal sperm concentration and plasma testosterone level significantly increased (P<0.05) in the ascorbic acid treated animals when compared to the control animals. The results suggest that ascorbic acid supplementation improves reproductive traits of male rats that are associated with high fertility. PMID- 15823362 TI - Taking measure of war trauma. PMID- 15823363 TI - Control of suffering on the slippery slope of care. PMID- 15823364 TI - Child survival: time to match commitments with action. PMID- 15823365 TI - Treating genetic disease through RNA interference. PMID- 15823366 TI - Ensuring the safety of HIV/AIDS generics. PMID- 15823367 TI - Matrix metalloproteinases biomarkers in multiple sclerosis. PMID- 15823368 TI - Apoptotic cell death: the caspase-cleavage "gold rush". PMID- 15823369 TI - Philip Landrigan: children's health crusader. PMID- 15823370 TI - Isolation of patients with MRSA infection. PMID- 15823371 TI - Isolation of patients with MRSA infection. PMID- 15823373 TI - Isolation of patients with MRSA infection. PMID- 15823374 TI - Cardiovascular risk assessment. PMID- 15823375 TI - Screening of organ and tissue donors for rabies. PMID- 15823376 TI - Misleading use of risk ratios. PMID- 15823377 TI - Cardiovascular risk assessment. PMID- 15823378 TI - Numbers needed to treat (needlessly?). PMID- 15823379 TI - Forest plots and the interpretation of subgroups. PMID- 15823380 TI - Prevalence of serious mental disorder in 7000 refugees resettled in western countries: a systematic review. AB - BACKGROUND: About 13 million people are classified as refugees worldwide, and many more former refugees have been granted citizenship in their new countries. However, the prevalence of post-traumatic stress disorder, major depression, or psychotic illnesses in these individuals is not known. We did a systematic review of surveys about these disorders in general refugee populations in western countries. METHODS: We searched for psychiatric surveys that were based on interviews of unselected refugee populations and that included current diagnoses of post-traumatic stress disorder, major depression, psychotic illnesses, or generalised anxiety disorder. We did computer-assisted searches, scanned reference lists, searched journals, and corresponded with authors to determine prevalence rates of these mental disorders and to explore potential sources of heterogeneity, such as diagnostic criteria, sampling methods, and other characteristics. FINDINGS: 20 eligible surveys provided results for 6743 adult refugees from seven countries, with substantial variation in assessment and sampling methods. In the larger studies, 9% (99% CI 8-10%) were diagnosed with post-traumatic stress disorder and 5% (4-6%) with major depression, with evidence of much psychiatric comorbidity. Five surveys of 260 refugee children from three countries yielded a prevalence of 11% (7-17%) for post-traumatic stress disorder. Larger and more rigorous surveys reported lower prevalence rates than did studies with less optimum designs, but heterogeneity persisted even in findings from the larger studies. INTERPRETATION: Refugees resettled in western countries could be about ten times more likely to have post-traumatic stress disorder than age matched general populations in those countries. Worldwide, tens of thousands of refugees and former refugees resettled in western countries probably have post traumatic stress disorder. PMID- 15823381 TI - Medical end-of-life decisions in neonates and infants in Flanders. AB - BACKGROUND: Paediatricians are increasingly confronted with end-of-life decisions in critically ill neonates and infants. Little is known about the frequency and characteristics of end-of-life decisions in this population, nor about the relation with clinical and patients' characteristics. METHODS: A death certificate study was done for all deaths of neonates and infants in the whole of Flanders over a 12 month period (August, 1999, to July, 2000). We sent an anonymous questionnaire by mail to the attending physician for each of the 292 children who died under the age of 1 year. Information on patients was obtained from national registers. An attitude study was done for all physicians who attended at least one death during the study period. FINDINGS: 253 (87%) of the 292 questionnaires were returned, and 121 (69%) of the 175 physicians involved completed the attitude questions. An end-of-life decision was possible in 194 (77%; 95% CI 70.4-82.4) of the 253 deaths studied, and such a decision was made in 143 cases (57%; 48.9-64.0). Lethal drugs were administered in 15 cases among 117 early neonatal deaths and in two cases among 77 later deaths (13%vs 3%; p=0.018). The attitude study showed that 95 (79%; 70.1-85.5) of the 121 physicians thought that their professional duty sometimes includes the prevention of unnecessary suffering by hastening death and 69 (58%; 48.1-66.5) of 120 supported legalisation of life termination in some cases. INTERPRETATION: Death of neonates and infants is commonly preceded by an end-of-life decision. The type of decision varied substantially according to the age of the child. Most physicians favour legalisation of the use of lethal drugs in some cases. PMID- 15823382 TI - Re-emergence of Chlamydia trachomatis infection after mass antibiotic treatment of a trachoma-endemic Gambian community: a longitudinal study. AB - BACKGROUND: Community-wide mass antibiotic treatment is a central component of trachoma control. The optimum frequency and duration of treatment are unknown. We measured the effect of mass treatment on the conjunctival burden of Chlamydia trachomatis in a Gambian community with low to medium trachoma prevalence and investigated the rate, route, and determinants of re-emergent infection. METHODS: 14 trachoma-endemic villages in rural Gambia were examined and conjunctival swabs obtained at baseline, 2, 6, 12, and 17 months. Mass antibiotic treatment with azithromycin was given to the community at baseline. C trachomatis was detected by qualitative PCR and individual infection load then estimated by real-time quantitative PCR. FINDINGS: C trachomatis was detected in 95 (7%) of 1319 individuals at baseline. Treatment coverage was 83% of the population (1328 of 1595 people). The effect of mass treatment was heterogeneous. In 12 villages all baseline infections (34 [3%] of 1062 individuals) resolved, and prevalence (three [0.3%]) and infection load remained low throughout the study. Two villages (baseline infection: 61 [24%] of 257 individuals) had increased infection 2 months after treatment (74 [30%]), after extensive contact with other untreated communities. Subsequently, this value reduced to less than half of that before treatment (25 [11%]). INTERPRETATION: Mass antibiotic treatment generally results in effective, longlasting control of C trachomatis in this environment. For low prevalence regions, one treatment episode might be sufficient. Infection can be reintroduced through contact with untreated populations. Communities need to be monitored for treatment failure and control measures implemented over wide geographical areas. PMID- 15823383 TI - Medical end-of-life decisions made for neonates and infants in the Netherlands, 1995-2001. AB - End-of-life decision-making for severely affected infants might be influenced by technical advances and societal debates. In 2001, we assessed the proportion of deaths of infants younger than 1 year that were preceded by end-of-life decisions, by replicating a questionnaire study from 1995. This proportion increased from 62% to 68% (weighted percentages), but the difference was not significant. Most of these decisions were to forgo life-sustaining treatment. Decisions to actively end the lives of infants not dependent on life-sustaining treatment remained stable at 1%. The practice of end-of-life decision-making in neonatology of 2001 has changed little since 1995. PMID- 15823384 TI - A twin with Cushing's disease. PMID- 15823385 TI - Type 2 diabetes: principles of pathogenesis and therapy. AB - Type 2 diabetes mellitus has become an epidemic, and virtually no physician is without patients who have the disease. Whereas insulin insensitivity is an early phenomenon partly related to obesity, pancreas beta-cell function declines gradually over time already before the onset of clinical hyperglycaemia. Several mechanisms have been proposed, including increased non-esterified fatty acids, inflammatory cytokines, adipokines, and mitochondrial dysfunction for insulin resistance, and glucotoxicity, lipotoxicity, and amyloid formation for beta-cell dysfunction. Moreover, the disease has a strong genetic component, but only a handful of genes have been identified so far: genes for calpain 10, potassium inward-rectifier 6.2, peroxisome proliferator-activated receptor gamma, insulin receptor substrate-1, and others. Management includes not only diet and exercise, but also combinations of anti-hyperglycaemic drug treatment with lipid-lowering, antihypertensive, and anti platelet therapy. PMID- 15823386 TI - Type 2 diabetes. PMID- 15823387 TI - Sample size calculations in randomised trials: mandatory and mystical. AB - Investigators should properly calculate sample sizes before the start of their randomised trials and adequately describe the details in their published report. In these a-priori calculations, determining the effect size to detect--eg, event rates in treatment and control groups--reflects inherently subjective clinical judgments. Furthermore, these judgments greatly affect sample size calculations. We question the branding of trials as unethical on the basis of an imprecise sample size calculation process. So-called underpowered trials might be acceptable if investigators use methodological rigor to eliminate bias, properly report to avoid misinterpretation, and always publish results to avert publication bias. Some shift of emphasis from a fixation on sample size to a focus on methodological quality would yield more trials with less bias. Unbiased trials with imprecise results trump no results at all. Clinicians and patients deserve guidance now. PMID- 15823388 TI - Pushing up smoking incidence: plans for a privatised tobacco industry in Moldova. AB - Moldova, one of the former Soviet republics and Europe's poorest country, has so far resisted pressure to privatise its tobacco industry. This paper examines the policies pursued by the transnational tobacco companies in Moldova in order to inform the ongoing debate about tobacco industry privatisation. We analysed relevant internal industry documents made public through litigation. The documents suggest that although a competitive tender for the state owned monopoly was later announced, British American Tobacco (BAT) and the German manufacturer Reemtsma each initially sought to secure a closed deal, with BAT accusing Reemtsma of underhand tactics. Imperial Tobacco, which now owns Reemstma, was unable to comment on these allegations as it only acquired Reemstma after the events in question. BAT sought to acquire a monopoly position, bolstered by excise rules developed by the company that would uniquely favour its products. Despite hoping to establish a monopoly, it planned intensive marketing, as if in a competitive market, aiming to target young urban dwellers, particularly opinion leaders. In so doing it predicted that smoking uptake would increase, especially among women. The documents also suggest that BAT was aware of the sensitive nature of its plans to cull the processing workforce and aimed to present "sanitised" information on future employment levels to the Moldovans. The potential for tobacco industry privatisation to undermine tobacco control and promote cigarette consumption is highlighted and is consistent with economic theory. Countries planning tobacco industry privatisation should ensure a transparent and competitive privatisation process, seek to prevent the predicted increase in consumption by implementing effective tobacco control policies and consider the impacts on employment. Multilateral financial organisations promoting tobacco industry privatisation could ensure their loan conditions protect public health by making the implementation of tobacco control policies a pre-requisite for privatisation. PMID- 15823389 TI - Lupin flour anaphylaxis. PMID- 15823390 TI - High molecular weight DNA recovery from soils prerequisite for biotechnological metagenomic library construction. AB - Soil is a complex environment considered as one of the main reservoirs of microbial diversity. However, the inability to cultivate most soil bacteria hampered fundamental attempts to determine the diversity of the prokaryotic world and limited its industrial exploitation. In the last 20 years, new methods have been developed to overcome these limitations based on the direct extraction of DNA from bacteria in their natural environment. In addition to fundamental research, the cloning of the extracted DNA for the development of metagenomic DNA clone libraries offers possibilities to discover novel bio-molecules through the expression of genes from uncultivated bacteria in surrogate bacterial hosts. However, such objectives require adapting DNA extraction methods and cloning strategies in order that entire gene clusters encoding biosynthetic pathway for secondary metabolites can be cloned. In this paper, we report that the size of DNA fragments extracted from soil varied in a range between less than 100 kb and more than 400 kb depending on the soil. The relatively limited size of DNA fragments extracted from some soil was not only due to mechanical, chemical or enzymatic shearing of the DNA during the extraction process but partly to the microbial growth status. Stimulating bacteria in situ by providing nutrients to the soil improved the size of extracted DNA, but it modified the bacterial community structure. PMID- 15823391 TI - Detection of antibiotic resistance genes in different Salmonella serovars by oligonucleotide microarray analysis. AB - In this study the feasibility of 50- and 60-mer oligonucleotides in microarray analysis for the detection and identification of antibiotic resistance genes in various Salmonella strains was assessed. The specificity of the designed oligonucleotides was evaluated, furthermore the optimal spotting concentration was determined. The oligonucleotide microarray was used to screen two sets of Salmonella strains for the presence of several antibiotic resistance genes. Set 1 consisted of strains with variant Salmonella Genomic Island 1 (SGI1) multidrug resistance (MDR) regions of which the antibiotic resistance profiles and genotypes were known. The second set contained strains of which initially only phenotypic data were available. The microarray results of the first set of Salmonella strains perfectly matched with the phenotypic and genotypic information. The microarray data of the second set were almost completely in concordance with the available phenotypic data. It was concluded that the microarray technique in combination with random primed genomic labeling and 50- or 60-mer oligonucleotides is a powerful tool for the detection of antibiotic resistance genes in bacteria. PMID- 15823392 TI - Application of a microplate scale fluorochrome staining assay for the assessment of viability of probiotic preparations. AB - Cell viability in probiotic preparations is traditionally assessed by the plate count technique. Additionally, fluorescent staining combined with epifluorescence microscopy or flow cytometry has been developed for the viability assessment, but the currently available assays are either laborious or require highly sophisticated equipment. The aim of this study was to investigate the applicability of a microplate scale fluorochrome assay for predicting the cell state of freeze-dried Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis preparations. In addition to viability assessment with LIVE/DEAD BacLight Bacterial Viability Kit, DiBAC(4)3 stain was used for the kinetic measurement of changes in bifidobacterial cell membrane functions during exposure to low pH. The microplate scale fluorochrome assay results on the viability and cell numbers of probiotic preparations correlated well with the results obtained with the culture based technique and (with few exceptions) with epifluorescence microscopy. The assay was applicable also for the viability assessment of stressed (acid-treated) cells provided that the cell density in treatments was adjusted to the optimal measurement level of the fluorometer. The microplate scale fluorochrome assay offers a rapid and robust tool for the viability assessment of probiotic preparations, and enables also kinetic measurements. PMID- 15823393 TI - Design and application of an oligonucleotide microarray for the investigation of compost microbial communities. AB - A microarray consisting of oligonucleotide probes targeting variable regions of the 16S rRNA gene was designed and tested for the investigation of microbial communities in compost. Probes were designed for microorganisms that have been previously reported in the composting process and for plant, animal and human pathogens. The oligonucleotide probes were between 17 and 25 bp in length and included mostly species-specific sequences. Validation of probe specificity and optimization of hybridization conditions were conducted using fluorescently labeled 16S rRNA gene PCR products of pure culture strains. A labeling method employing a Cy3 or Cy5-labeled forward primer together with a phosphate conjugated reverse primer for the production of single stranded DNA after a digestion step was optimised and used to label target DNA. A combination of two different DNA extraction methods using both physical and chemical lysis was found to give the best DNA yields. Increased hybridization signal intensities were obtained for probes modified with a 12 mer T-spacer. The microarray was found to have a detection limit of 10(3) cells, although in compost spiking experiments, the detection limit was reduced to 10(5) cells. The application of the microarray to compost samples indicated the presence of Streptococcus, Acinetobacter lwoffii, and Clostridium tetani in various compost samples. The presence of A. lwoffii in those compost samples was confirmed by PCR using primers specific for the organism. The aim of this study was to develop a molecular tool that would allow screening for the presence or absence of different microorganisms within compost samples. PMID- 15823394 TI - An oligonucleotide microarray for the identification and differentiation of trichothecene producing and non-producing Fusarium species occurring on cereal grain. AB - Cereal grain may be infected with a number of Fusarium species some of which are producers of highly toxic compounds such as the trichothecenes. Correct identification of these species is essential for risk assessment of cereal grain for human or animal consumption. Most of the available methods for identification are either time consuming or aimed at only one or a few target species. Microarray technology offers parallel analysis of a high number of DNA targets. In this study 57 capture oligonucleotides (CO) were designed based upon Fusarium ITS2 rDNA sequences, and used for microarray production. From this array COs could be selected that were able to hybridise specifically to labelled PCR products from the ITS region of Fusarium graminearum/Fusarium culmorum, Fusarium pseudograminearum, Fusarium poae, Fusarium sporotrichioides, Fusarium equiseti, Fusarium langsethiae and Fusarium tricinctum/Fusarium avenaceum. A few COs showed some cross hybridisation to non-target species. In a preliminary experiment it was shown that this cross hybridisation could be eliminated by increasing hybridisation stringency. The array could be used to detect individual Fusarium species in mixed samples and in environmental samples. This study demonstrates the feasibility of oligonucleotide microarrays for parallel detection of a number of Fusarium species. PMID- 15823395 TI - Evaluation of DNA extraction methods from mouse stomachs for the quantification of H. pylori by real-time PCR. AB - Real-time PCR methods have recently been developed for the quantification of Helicobacter pylori from infected mouse stomachs. However, the extent to which results is affected by the efficiency of different methods of DNA extraction and the degree of inhibition of the subsequent PCR have largely been ignored. In this study, mouse stomachs were processed using two homogenisation methods: complete disruption using a blender and homogenisation by vortexing with glass beads. Each procedure was followed by DNA purification by three different protocols-two commercially available kits-Qiagen DNA Mini Tissue kit and Qiagen Stool Kit and a phenol-chloroform extraction method. PCR inhibition was assessed by screening for mouse DNA and for H. pylori DNA after spiking stomach extracts with H. pylori 16S rDNA. PCR inhibition was found to be lower in DNA samples prepared by vortexing and processed by column kits. Validation of procedures was performed by quantification of H. pylori DNA and mouse DNA in infected mouse stomachs. Homogenisation with glass beads followed by the Qiagen Tissue kit was found to be the most suitable protocol combining high extraction and detection efficiency of 16S rDNA in the presence of a mouse DNA background. PMID- 15823396 TI - Comparison of NaOH-N-acetyl cysteine and sulfuric acid decontamination methods for recovery of mycobacteria from clinical specimens. AB - We compared the NaOH-N-acetyl cysteine (NaOH-NALC) and the sulfuric acid decontamination procedure in the detection of mycobacteria using the Mycobacteria Growth Indicator Tube (MGIT). In total 219 sputum specimens were collected from 142 Zambian patients and subjected to mycobacterial culture. One half of the specimen was decontaminated with NaOH-NALC and the other half was decontaminated with sulfuric acid. From the 438 samples a total of 261 (60%) cultures yielded growth of mycobacteria, consisting of 22 different species. The sulfuric acid method was more successful than the NaOH-NALC method in recovering mycobacteria in MGITs (146 versus 115 respectively, p = 0.001). Of the 146 positive mycobacterial cultures recovered after sulfuric acid decontamination 28 were Mycobacterium tuberculosis, 84 nontuberculous mycobacteria (NTM) and 34 acid fast bacterial isolates which could not be identified to the species level. The 115 mycobacteria recovered by the NaOH-NALC method consisted of 34 M. tuberculosis strains, 55 NTM and 26 acid fast bacteria that could not be identified. The most frequently isolated NTM were Mycobacterium lentiflavum and Mycobacterium intracellulare. Comparing the two decontamination methods the recovery of NTM in the sulfuric acid group was significant higher than in the NaOH-NALC group (p = 0.001). In contrast, no significant difference was found for the recovery of M. tuberculosis. These results show that the decontamination method used affects the recovery of nontuberculous mycobacteria in particular. PMID- 15823397 TI - Assessment of self-organizing maps to analyze sole-carbon source utilization profiles. AB - The use of community-level physiological profiles obtained with Biolog microplates is widely employed to consider the functional diversity of bacterial communities. Biolog produces a great amount of data which analysis has been the subject of many studies. In most cases, after some transformations, these data were investigated with classical multivariate analyses. Here we provided an alternative to this method, that is the use of an artificial intelligence technique, the Self-Organizing Maps (SOM, unsupervised neural network). We used data from a microcosm study of algae-associated bacterial communities placed in various nutritive conditions. Analyses were carried out on the net absorbances at two incubation times for each substrates and on the chemical guild categorization of the total bacterial activity. Compared to Principal Components Analysis and cluster analysis, SOM appeared as a valuable tool for community classification, and to establish clear relationships between clusters of bacterial communities and sole-carbon sources utilization. Specifically, SOM offered a clear bidimensional projection of a relatively large volume of data and were easier to interpret than plots commonly obtained with multivariate analyses. They would be recommended to pattern the temporal evolution of communities' functional diversity. PMID- 15823398 TI - The use of RAPD genomic fingerprinting to study relatedness in strains of Acidithiobacillus ferrooxidans. AB - Twelve strains of Acidithiobacillus ferrooxidans were recovered from acid mine drainage (AMD) sites from three different geographical locations: Copper Cliff, Ontario, Canada; Mineral City, OH, USA; and Cornwall, England. The spread-plate technique and various culture media were used to isolate and purify all strains. DNA was extracted from each purified culture and amplified using PCR and twenty, 10-mer primers. Amplification products were separated by gel electrophoresis and photographed under UV light. The RAPD (Randomly Amplified Polymorphic DNA) profiles were compared on the basis of the presence or absence of each DNA band and a data matrix was constructed. Strain diversity was calculated using the Jaccard's coefficient and UPGMA (Unweighted Pair-Group Arithmetic Average Clustering) cluster analysis. The variations in the banding patterns indicated genomic variability among the twelve A. ferrooxidans strains tested. The primers used in this study grouped the twelve strains into five major groups. Similarities between the strains ranged from 5.49% to 85.14%. These results show that the strains have a high degree of genomic diversity and that the RAPD procedure is a powerful technique to assess strain variability in this bacterium. PMID- 15823399 TI - A Pyrosequencing assay for rapid recognition of SNPs in Mycobacterium tuberculosis embB306 region. AB - Ethambutol (EMB) is in use worldwide as a first-line anti-tuberculosis drug and substitutions in codon 306 of the embB gene are the most common mutations found in EMB resistant Mycobacterium tuberculosis (MTB) strains. Pyrosequencing is a real time sequencing method able to rapidly detect mutations in a large number of samples. Using this technique we analyzed, in parallel with conventional sequencing, a 24 bp region of the embB gene of 28 MTB clinical isolates. Pyrosequencing efficiently identified all embB306 mutations, detecting three different single-base substitutions leading to 2 amino acid changes (Met to Val or Ile). Mutated embB alleles were detected in 2 multidrug-resistant (MDR) EMB susceptible strains. Overall, our results demonstrated that the Pyrosequencing method efficiently recognizes mutations in embB in a very short time and represents a valid molecular method to detect mutations in the MTB embB306 region. PMID- 15823400 TI - Bioactivity quantification of crude bacteriocin solutions. AB - Bacteriocin activity can be quantified by a simple parallel line model based on an agar diffusion assay. The quotient between the inhibition zone area and the sensitivity of the indicator bacterium can be used to compare different bacteriocins. Deferred calculations of relative potencies are possible, once purified compounds are obtained. PMID- 15823401 TI - Rapid protocol for electroporation of Clostridium perfringens. AB - A rapid and simple method has been developed for the electroporation of Clostridium perfringens with plasmid DNA. The new improvements, harvesting cells early in the logarithmic stage of growth, keeping the cells at room temperature and the absence of post-shock incubation on ice increased transformation efficiency by one order of magnitude. PMID- 15823402 TI - Enhanced swarming of bacteria on agar plates containing the surfactant Tween 80. AB - A widely used method for quantifying swarming motility is the swarm plate assay. A significant increase in the motility halo size formed by Escherichia coli or Azospirillum brasilense was measured on Tween 80-containing agar relative to untreated agar. This improvement could benefit the identification of mutants in swarming motility. PMID- 15823403 TI - Influence of DNA stiffness in protein-DNA recognition. AB - Protein-DNA recognition plays an essential role in the regulation of gene expression. The protein-DNA binding specificity is based on direct atomic contacts between protein and DNA and/or the conformational properties of DNA. In this work, we have analyzed the influence of DNA stiffness (E) to the specificity of protein-DNA complexes. The average DNA stiffness parameters for several protein-DNA complexes have been computed using the structure based sequence dependent stiffness scale. The relationship between DNA stiffness and experimental protein-DNA binding specificity has been brought out. We have investigated the importance of DNA stiffness with the aid of experimental free energy changes (DeltaDeltaG) due to binding in several protein-DNA complexes, such as, ETS proteins, 434, lambda, Mnt and trp repressors, 434 cro protein, EcoRV endonuclease V and zinc fingers. We found a correlation in the range 0.65 0.97 between DeltaDeltaG and E in these examples. Further, we have qualitatively analyzed the effect of mutations in the target sequence of lambda repressor and we observed that the DNA stiffness could correctly identify 70% of the correct bases among the considered nine positions. PMID- 15823404 TI - Genomic peculiarity of coding sequences and metabolic potential of probiotic Escherichia coli strain Nissle 1917 inferred from raw genome data. AB - Probiotic Escherichia coli strain Nissle 1917 (O6:K5:H1) is a commensal E. coli isolate that has a long tradition in medicine for the treatment of various intestinal disorders in humans. To elucidate the molecular basis of its probiotic nature, we started sequencing the genome of this organism with a whole-genome shotgun approach. A 7.8-fold coverage of the genomic sequence has been generated and is now in the finishing stage. To exploit the genome data as early as possible and to generate hypotheses for functional studies, the unfinished sequencing data were analyzed in this work using a new method [Sun, J., Zeng, A.P., 2004. IdentiCS--identification of coding sequence and in silico reconstruction of the metabolic network directly from unannotated low-coverage bacterial genome sequence. BMC Bioinformatics 5, 112] which is particularly suitable for the prediction of coding sequences (CDSs) from unannotated genome sequence. The CDSs predicted for E. coli Nissle 1917 were compared with those of all five other sequenced E. coli strains (E. coli K-12 MG1655, E. coli K-12 W3110, E. coli CFT073, EHEC O157:H7 EDL933 and EHEC O157:H7 Sakai) published to date. Five thousand one hundred and ninety-two CDSs were predicted for E. coli Nissle 1917, of which 1065 were assigned with enzyme EC numbers. The comparison of all predicted CDSs of E. coli Nissle 1917 to the other E. coli strains revealed 108 CDSs specific for this isolate. They are organized as four big genome islands and many other smaller gene clusters. Based on CDSs with EC numbers for enzymes, the potential metabolic network of Nissle 1917 was reconstructed and compared to those of the other five E. coli strains. Overall, the comparative genomic analysis sheds light on the genomic peculiarity of the probiotic E. coli strain Nissle 1917 and is helpful for designing further functional studies long before the sequencing project is completely finished. PMID- 15823405 TI - Standardization strategy for quantitative PCR in human seminoma and normal testis. AB - Housekeeping genes are commonly used as endogenous references in quantitative RT PCR. Ideally these genes are constitutionally expressed by all cell types and do not vary under experimental conditions. Tissues of 9 normal testes and 22 classical pure seminoma were obtained for RNA-extraction. Real-time RT-PCR was used to examine the mRNA-expression of ubiquitin C, beta-actin, GAPDH, 18S ribosomal RNA (18S rRNA) and porphobilinogen-deaminase (PBGD). Additionally, 3 normal testicular tissues and 39 seminoma, including 1 normal testis and 17 seminoma of the RT-PCR group, were utilized for microarray analyses. Ubiquitin C (protein degradation) was down-regulated, GAPDH (carbohydrate metabolism), beta actin (cytoskeleton), 18S rRNA (ribosome) and PBGD (porphyrin metabolism) were up regulated in seminoma. A normalization of the target gene data with up-regulated housekeeping genes would equalize or underestimate up-regulated data and overestimate down-regulated data. We demonstrate that none of the investigated housekeeping genes is suitable for normalization of the target gene RT-PCR data, but may be essential for tumor metabolism in human seminoma. Further, we developed a standardization strategy, which is applicable to many experimental investigations. PMID- 15823406 TI - A rapid real-time qRT-PCR assay for ovine beta-actin mRNA. AB - Beta-Actin mRNA is often used for normalization in gene expression experiments. We describe a sensitive, rapid and specific quantitative assay for the cytoplasmic ovine beta-actin mRNA. The assay was based on the polymerase chain reaction (PCR) with real-time fluorescence resonance energy transfer (FRET) measurements to amplify cDNA products reverse transcribed from mRNA. A part of the ovine beta-actin sequence was amplified from cDNA from fetal ovine synovial (FOS) cells with mRNA-specific primers and cloned into a plasmid clone. The assay standard curve was constructed with dilutions of this plasmid. The assay was linear over five orders of magnitude and detected down to 600 copies per reaction of target DNA. Intraassay coefficient of variation was 12%. Detection of the beta actin gene was eliminated by designing FRET probes at splice junctions and detection of putative processed pseudogenes was minimized by using FRET assay design with four oligonucleotides. We measured 0.2 copies per cell in RNA preparations without reverse transcription and DNase digestion. This might represent processed pseudogenes. In constrast, we measured 1400 beta-actin mRNA copies per cell in RNA preparations after the RT and DNase steps. The assay should, therefore, be sensitive enough to measure beta-actin from a single individual cell. Dilution of target DNA in murine RNA or ovine cDNA preparations did not effect efficiency of PCR or linearity of the assay. The quantitative assay described in this work can be used to correct for variations in various real-time qRT-PCR experiments in ovine cells with diverse goals, including gene expression studies, quantitation of viral load in infected cells and in various gene therapy experiments measuring vector load and expression in transduced cells. PMID- 15823407 TI - Production in Saccharomycescerevisiae of MS2 virus-like particles packaging functional heterologous mRNAs. AB - Recently, DNA bacteriophages (M13, lambda) have been genetically engineered to transfer genes into mammalian cells. Although efficiencies observed are still relatively low, this opens the possibility of using these viruses as a new class of transfection agents not only for fundamental research purposes but also in gene therapy protocols or in other applications like vaccination. In this respect, it has been shown that a lambda bacteriophage engineered to express the hepatitis B surface antigen in mammalian cells could elicit an immune response against this antigen in mice and rabbits without any specific targeting of the bacteriophage. These impressive results would be even more encouraging if they could be obtained with an RNA bacteriophage, as RNA vaccines are preferred over DNA vaccines for safety reasons. Up to now, RNA bacteriophages have never been engineered for gene delivery. In this paper, we have sought to determine whether such a vector could be obtained by engineering the RNA bacteriophage MS2. We show that MS2 can be produced as virus-like particles (VLPs) in Saccharomyces cerevisiae and is able to package functional heterologous mRNAs, provided that these mRNAs contain the MS2 packaging sequence. For instance, linking the MS2 packaging sequence to the human growth hormone (hGH) mRNA enabled the packaging of this particular mRNA in MS2 VLPs. Functionality in eukaryotic systems of packaged mRNAs was confirmed by showing that mRNAs purified from VLPs can be efficiently translated in vitro and in cell cultures. The high stability of MS2 could, therefore, make MS2 VLPs a very powerful carrier for RNA vaccines. PMID- 15823408 TI - Novel interface-binding chloroperoxidase for interfacial epoxidation of styrene. AB - A unique interface-binding chloroperoxidase (CPO) was developed and examined for interfacial biocatalysis. Native CPO was conjugated with polystyrene (PS) to form a surfactant-like structure that self assembled at oil-water interfaces. While enantioselectivity of the enzyme was maintained, the interfacial assembly of the enzyme improved its overall catalytic efficiency as compared to that observed with the enzyme contained in the bulk aqueous phase. The PS conjugated CPO (PS CPO) showed a 2.5-fold enhancement of enzyme productivity versus native CPO under batch reaction conditions for the epoxidation of styrene, whereas a 25-fold improvement was realized in a continuous feeding reaction to reach a productivity of 10 micromol h-1 mg protein-1. The interface-binding enzyme also demonstrated several other advantages such as suppressing unwanted side reactions including the hydrolysis of styrene epoxide products, stabilizing the enzyme by limiting its exposure to both the oxidant H2O2 and epoxide products, and alleviating the deactivating effect of interfacial stress on enzymes by functioning as a surfactant. PMID- 15823409 TI - New effective sources of the Staphylococcus simulans lysostaphin. AB - The gene encoding Staphylococcus simulans lysostaphin has been cloned into two Escherichia coli expression systems: pET23b+ (Novagen, UK) and pBAD/Thio-TOPO (Invitrogen, USA), which allow the overexpression of a target protein as a fusion protein. The enzyme produced in the pET system contains a cluster of six histidines at the C-terminus, and the protein produced in the pBAD system contains 133 additional amino acid residues at the N-terminus, including thioredoxin, a cluster of six histidines and a recognition site for endoprotease Factor Xa. The recombinant enzymes were purified by metal-affinity chromatography on a Co2+-Sepharose column. Approximately 20 mg of purified recombinant enzyme were obtained in the pET expression system and 39 mg in the pBAD system, from a 1 L culture. The obtained fusion protein from the pET system revealed specific activity that was approximately 10 times higher than that of the fusion protein from the pBAD system (970 U/mg versus 83 U/mg). The purified enzymes displayed maximum activity at close to 45 degrees C and pH 8.0 or 7.5 for the enzyme obtained from pET and pBAD system, respectively. The lysostaphin activity was strongly inhibited by Zn2+ or Cu2+ (2 mM) with a 70-80% decrease. The Ni2+ (2 mM) also inhibited the enzyme with a 60 and 20% activity decrease for enzyme from the pET and pBAD system, respectively. The Co2+ had no impact on enzymatic activity at the 2 mM concentration; however, 30 and 20% activity decreases were observed at the 10mM concentration for the enzyme obtained from the pET and pBAD expression systems, respectively. EDTA, known as a strong inhibitor of the native lysostaphin, had no impact on the antistaphylococcal activity of either recombinant enzyme. PMID- 15823410 TI - In silico screening of drug databases for TSE inhibitors. AB - Today, thousands of different chemical compounds are used as drugs for a wealth of different indications. Here, we demonstrate the use of a novel conformational drug database for the search of compounds with a positive influence on Transmissible Spongiform Encephalopathies (TSEs) by using two- and three dimensional structural similarity to compounds with known effect. Both methods are combined to deduce a list of 16 candidate drugs. The proposal of a small number of putative inhibitors out of about 2300 approved essential drugs allows testing by expensive or time-consuming methods with the advantage that all agents are well known and suitable for use in humans. PMID- 15823411 TI - Synonymous codon usage and gene function are strongly related in Oryza sativa. AB - The relationship between codon usage and gene function was investigated while considering a dataset of 2106 nuclear genes of Oryza sativa. The results of standard chi(2) test and F-statistic showed that for every 59 synonymous codons, a strongly significant association with gene functional categories existed in rice, indicating that codon usage was generally coordinated with gene function whether it was at the level of individual amino acids or at the level of nucleotides. However, it could not be directly said that the use of every codons differed significantly between any two functional categories. Notably, there existed large difference both in selection for biased codons or selection intensity among functional categories. Therefore, we identified at least two classes of genes: one group of genes, mainly belonging to the "METABOLISM" category, was tended to use G- and/or C-ending codons while the other was more biased to choose codons ending with A and/or U. The latter group contained genes of various functions, especially those genes classified into the "Nuclear Structure" category. These observations will be more important for molecular genetic engineering and genome functional annotation. PMID- 15823412 TI - Computational multiple steady states for enzymatic esterification of ethanol and oleic acid in an isothermal CSTR. AB - The capacity of complex biochemical reaction networks (consisting of 11 coupled non-linear ordinary differential equations) to show multiple steady states, was investigated. The system involved esterification of ethanol and oleic acid by lipase in an isothermal continuous stirred tank reactor (CSTR). The Deficiency One Algorithm and the Subnetwork Analysis were applied to determine the steady state multiplicity. A set of rate constants and two corresponding steady states are computed. The phenomena of bistability, hysteresis and bifurcation are discussed. Moreover, the capacity of steady state multiplicity is extended to the family of the studied reaction networks. PMID- 15823413 TI - Identification of conserved regulatory RNA structures in prokaryotic metabolic pathway genes. AB - A combination of algorithms to search RNA sequence for the potential for secondary structure formation, and search large numbers of sequences for structural similarity, were used to search the 5'UTRs of annotated genes in the Escherichia coli genome for regulatory RNA structures. Using this approach, similar RNA structures that regulate genes in the thiamin metabolic pathway were identified. In addition, several putative regulatory structures were discovered upstream of genes involved in other metabolic pathways including glycerol metabolism and ethanol fermentation. The results demonstrate that this computational approach is a powerful tool for discovery of important RNA structures within prokaryotic organisms. PMID- 15823414 TI - Reverse engineering of biochemical equations from time-course data by means of genetic programming. AB - Increased research aimed at simulating biological systems requires sophisticated parameter estimation methods. All current approaches, including genetic algorithms, need pre-existing equations to be functional. A generalized approach to predict not only parameters but also biochemical equations from only observable time-course information must be developed and a computational method to generate arbitrary equations without knowledge of biochemical reaction mechanisms must be developed. We present a technique to predict an equation using genetic programming. Our technique can search topology and numerical parameters of mathematical expression simultaneously. To improve the search ability of numeric constants, we added numeric mutation to the conventional procedure. As case studies, we predicted two equations of enzyme-catalyzed reactions regarding adenylate kinase and phosphofructokinase. Our numerical experimental results showed that our approach could obtain correct topology and parameters that were close to the originals. The mean errors between given and simulation-predicted time-courses were 1.6 x 10(-5)% and 2.0 x 10(-3)%, respectively. Our equation prediction approach can be applied to identify metabolic reactions from observable time-courses. PMID- 15823415 TI - A simple mathematical model of cooperativity in receptor mosaics based on the "symmetry rule". AB - The phenomenon of receptor-receptor interactions was hypothesized about 20 years ago. It has been demonstrated by now that receptor-receptor interactions between G-protein coupled receptors (GPCRs) occur at plasma membrane level and result in the reciprocal modulation of their binding characteristics (i.e., cooperativity). One of the most important feature of this phenomenon is the concept of cluster of receptors, or receptor mosaic (RM). However, no proper mathematical approach has still been available to characterize RMs as far as their receptor composition, receptor topography and order of receptor activation inside the RM. This paper tries to fill the gap. A simple mathematical approach to the cooperativity in RMs formed by dimers of identical receptors and/or by iso-receptors is proposed. To this aim the so-called "symmetry rule" has been considered. This approach allows to describe by means of a simple energy function the effects of receptor composition (number of dimers), spatial organisation (respective location of the dimers) and order of activation (order according to which the single receptors are ligated) on the integrative cooperativity (index) of the RMs. PMID- 15823416 TI - The origin of the genetic code: theories and their relationships, a review. AB - A review of the main theories proposed to explain the origin of the genetic code is presented. I analyze arguments and data in favour of different theories proposed to explain the origin of the organization of the genetic code. It is possible to suggest a mechanism that makes compatible the different theories of the origin of the code, even if these are based on a historical or physicochemical determinism and thus appear incompatible by definition. Finally, I discuss the question of why a given number of synonymous codons was attributed to the amino acids in the genetic code. PMID- 15823417 TI - The effect of species models on estimates of within-lineage variation in integration. AB - Species may be modeled as comprised of individuals, populations or a virtual code. A virtual code can be understood as general potential that appears as actualization within specific environmental, both internal and external, contexts. These general potentials form a capacity to network that allows potentials to be expressed and offers robustness through its interconnections. In the present work, the degree of within-lineage variation in integration was not strongly model-dependent. However, the relationships among model-dependent estimates of such variation and within-lineage phyletic variation were not equal. The strongest relationship was between within-lineage variation in integration, when species were modeled as a virtual code, and within-lineage phyletic variation. The second strongest, and only other statistically significant relationship, was between variation in integration when species were modeled as a virtual code and as a collection of populations. The last result argues for a strong ontogenetic and micro-environmental effect on the expression of features in an individual. If species were a virtual code they would evolve by incorporation of all attributes, ontogenetic, environmental and genetic into that code until it becomes unstable and bifurcates. Species as a virtual code, an approach that explicitly incorporates developmental change into evolution, is a non-material representation of species as a complex information system, incorporating, if we refer to mathematical analysis, both the real and the imaginary. If one wished to stress the material, this study could be seen as empirical documentation of species as information systems. PMID- 15823418 TI - Higher mutation rate helps to rescue genes from the elimination by selection. AB - Directional mutation pressure associated with replication processes is the main cause of the asymmetry between the leading and lagging DNA strands in bacterial genomes. On the other hand, the asymmetry between sense and antisense strands of protein coding sequences is a result of both mutation and selection pressures. Thus, there are two different ways of superposition of the sense strand, on the leading or lagging strand. Besides many other implications of these two possible situations, one seems to be very important - because of the asymmetric replication-associated mutation pressure, the mutation rate of genes depends on their location. Using Monte Carlo methods, we have simulated, under experimentally determined directional mutation pressure, the divergence rate and the elimination rate of genes depending on their location in respect to the leading/lagging DNA strands in the asymmetric prokaryotic genome. We have found that the best survival strategy for the majority of genes is to sometimes switch between DNA strands. Paradoxically, this strategy results in higher substitution rates but remains in agreement with observations in bacterial genomes that such inversions are very frequent and divergence rate between homologs lying on different DNA strands is very high. PMID- 15823419 TI - Modeling the effect of acylated homoserine lactone antagonists in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is a gram-negative bacterium that causes serious illnesses, particularly in immunocompromised individuals, often with a fatal outcome. The finding that the acylated homoserine lactone quorum sensing (QS) system controls the production of virulence factors in P. aeruginosa makes this system a possible target for antimicrobial therapy. It has been suggested that an N-(3-oxododecanoyl)-homoserine lactone (3O-C12-HSL) antagonist, a QS blocker (QSB), would interfere efficiently with the quorum sensing system in P. aeruginosa and thus reduce the virulence of this pathogen. In this work, a mathematical model of the QS system in P. aeruginosa has been developed. The model was used to virtually add 3O-C12-HSL antagonists that differed in their affinity for the receptor protein and for their ability to mediate degradation of the receptor. The model suggests that very small differences in these parameters for different 3O-C12-HSL antagonists can greatly affect the success of QSB based inhibition of the QS system in P. aeruginosa. Most importantly, it is proposed that the ability of the 3O-C12-HSL antagonist to mediate degradation of LasR is the core parameter for successful QSB based inhibition of the QS system in P. aeruginosa. Finally, this study demonstrates that QSBs can shift the system to a low steady state, corresponding to an uninduced state and thus, suggests that the use of 3O-C12-HSL antagonists may constitute a promising therapeutic approach against P. aeruginosa involved infections. PMID- 15823420 TI - Neuronal nitric oxide synthase inhibition differentially affects oxytocin and vasopressin secretion in salt loaded rats. AB - Nitric oxide, an endogenous gas produced by nitric oxide synthase (NOS), has been described as a neuromodulator of hormone secretion, including the neurohypophysial peptides oxytocin (OT) and vasopressin (AVP), hormones involved in the sodium and water homeostasis. The presence of NOS in the hypothalamic nuclei as well as in the circumventricular organs suggests a nitrergic regulation of OT and AVP secretion. Thus, the aim of this study was to evaluate the effect of 7-nitroindazole (7-NI), a selective inhibitor of neuronal NOS, in the plasma OT and AVP levels in rats submitted to a short and long-term salt loading. We also evaluated the NOS activity in the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. Our data showed an increase of plasma OT and AVP levels in both short and long-term salt loading. The augment of plasma OT and AVP levels was accompanied by an increase of NOS activity in the SON and PVN. The injection of 7-NI potentiated the increase of plasma OT induced by salt loading, but inhibited the increase of plasma AVP in the same experimental conditions. These results indicate that, under short and prolonged osmotic stimulation, nitric oxide may differentially control the neurohypophysial secretion. PMID- 15823421 TI - The SNAP-25 gene may be associated with clinical response and weight gain in antipsychotic treatment of schizophrenia. AB - The synaptosomal-associated protein of 25 kDa (SNAP-25) is an essential component of the core complex that mediates presynaptic vesicle trafficking. Thus, SNAP-25 is directly involved in the release of neurotransmitters. Quantitative alterations of SNAP-25 expression have been reported in brain regions and cerebrospinal fluid (CSF) of schizophrenics and in haloperidol treated rats. This observed altered expression may be influenced by genetic variants of SNAP-25. We hypothesized that polymorphisms of the SNAP-25 gene (sites DdeI, MnlI and TaiI in the 3'UTR) are associated with antipsychotic drug response and induced weight gain. A sample of 59 patients with prior suboptimal response to antipsychotic treatment and diagnosed with DSM-IV schizophrenia or schizoaffective disorder was examined. Patients were administered clozapine, haloperidol, olanzapine or risperidone for up to 14 weeks. Clinical response was defined as the difference between the baseline and the endpoint total scores on the Positive and Negative Syndrome Scale (PANSS). Weight was assessed at baseline and at study endpoint. ANOVA revealed that the MnlI and TaiI polymorphisms were associated with response (F[2,53] = 4.57, p = 0.01 and F[2,52] = 3.53, p = 0.03) and with weight gain (F[2,52] = 4.28, p = 0.01 and F[2,51] = 3.38, p = 0.04). When covariates were included, the MnlI polymorphism remained significantly associated with changes of PANSS scores, but not with weight gain. The DdeI polymorphism was not associated with response or weight gain. These findings suggest that SNAP-25 gene variants affect clinical response in patients with prior poor response to antipsychotics. Weight changes do not seem to be associated with polymorphism of the SNAP-25 gene, however, replication in independent samples is warranted. PMID- 15823422 TI - Different regions of the mouse nestin enhancer may function differentially in nestin expression in an NSC-like cell line and astrocytes. AB - Nestin is a characteristic intermediate filament protein expressed in neural stem cells (NSCs). Evidence has shown that it is also found in reactive astrocytes. Previous studies have demonstrated that the second intron of the human and rat nestin genes harbors the central nervous system (CNS) enhancer and the midbrain enhancer, which regulate nestin expression in different regions of CNS during development. In this study, using an NSC-like cell line C17.2 and primarily cultured astrocytes, we show that both C17.2 cells and astrocytes express nestin. To characterize the nestin enhancer in further detail, we cloned the second intron of the mouse nestin gene, which is homologous to the human and rat counterparts as shown by DNA sequencing. Reporter assay indicated that the full length nestin enhancer was active in both C17.2 cells and astrocytes, consistent with the immunocytochemistry results. However, in C17.2 cells, the enhancer activity was attributed to the highly conserved 3' part, and the 5' part of the enhancer was suppressive to the transcription activation activity of the full length enhancer. While in astrocytes, both 3' and 5' parts were able to enhance the reporter gene expression. Our data suggested that different regions of the nestin enhancer might have different functions in C17.2 cells and astrocytes: while the 3' region activates transcription in both cell types, the 5' region suppresses in C17.2 cells but activates in astrocytes nestin expression. PMID- 15823424 TI - Developmental expression of 5-HT 5A receptor mRNA in the rat brain. AB - In the central nervous system, serotonin (5-HT) may function as a mitogen as well as a neurotransmitter; and its early appearance suggests a potential role in development. The present experiments were done to determine the localization of the mRNA coding for the 5-HT 5A receptor during development of the rat brain. 5 HT 5A gene transcription was assessed by in situ hybridization, from E18 and during postnatal (PN) development. An intense signal of 5-HT 5A mRNA was found in the cerebral cortex and olfactory nucleus at E18, PN0 and PN5. A sharp decrease at PN11 was followed by an increase until reaching the adult level in the cerebral cortex; whereas in the olfactory nucleus, transcription remained weak. In contrast, in the hippocampal formation the signal was weak in the CA1, CA2 and CA3 regions at E18 and PO; increased at P5 and then decreased at P11 before attaining the adult level. We conclude that the gene coding for the 5-HT 5A receptor is already active in the embryonic rat brain and is differentially expressed during development. PMID- 15823423 TI - A role for hypoxia-inducible factor-1alpha in desferoxamine neuroprotection. AB - The newborn brain has increased vulnerability to hypoxia-ischemia from maturational differences in the oxidative stress response. We hypothesized that desferoxamine (DFO), an iron chelator, would provide protection in an in vitro model of ischemia in part through activation of the hypoxia-inducible gene hypoxia-inducible factor-1alpha (HIF-1alpha). Hippocampal neurons from E16 CD1 mice were exposed to 3 h of oxygen and glucose deprivation with and without pretreatment with 10 mmol/L DFO in the presence and absence of 2 micromol/L antisense oligonucleotides specific for HIF-1alpha (antiHIF-1alpha). DFO pretreatment resulted in 45% reduction in cell death (p = 0.006). This protection was diminished with transfection of antiHIF-1alpha (p = 0.049). Blocking HIF 1alpha reduces DFO protection suggesting that DFO protects through iron chelation and HIF-1alpha induction. PMID- 15823425 TI - Effects of (+/-)-huprine Y and (+/-)-huprine Z, two new anticholinesterasic drugs, on muscarinic receptors. AB - The cholinergic profile of (+/-)-huprine Y and (+/-)-huprine Z on muscarinic receptors has been determined. Displacement of [3H]-pirenzepine and [3H]-QNB plus pirenzepine was performed in rat hippocampus. Both compounds showed a higher degree of affinity to M1 muscarinic receptors (P < 0.01) than to M2 muscarinic receptors. To determine the M1 agonist or antagonist role of the two huprines, studies of inositol phosphates (IP) production were performed. Both huprines significantly stimulated IP accumulation in a concentration-dependent manner. The reversion of this effect by different antagonists showed that M1 muscarinic receptors were activated by (+/-)-huprine Y and (+/-)-huprine Z, but some other mechanisms, such as alpha1-adrenoceptors or nicotinic receptors, were involved. PMID- 15823426 TI - High frequency activities in the human orbitofrontal cortex in sleep-wake cycle. AB - We recorded human orbitofrontal electrocorticogram during wakefulness and sleep in epileptic patients using subdural electrodes. During wakefulness and rapid eye movement (REM) sleep, we observed beta activity in the raw orbitofrontal signals. Power spectral analysis demonstrated beta enhancement during wakefulness and REM sleep when compared to slow wave sleep (SWS). During the phasic REM periods, the beta power was significantly lower than during the tonic REM periods. Gamma enhancement manifested itself in four out of six subjects during the phasic periods. This study is the first that has focused on electrical activity in the human orbitofrontal cortex. Although the role of the orbitofrontal cortex during sleep still remains unclear, high frequency activities give us important suggestions in elucidating the human sleep mechanism. PMID- 15823427 TI - Involvement of CD45 in central nervous system myelination. AB - Myelin is a multi-layered membranous lipid insulator surrounding axons that allows the rapid conduction of neuronal impulses. In the central nervous system (CNS), myelin is produced by oligodendrocytes. During development, morphologically immature oligodendrocyte precursor cells (OPCs) arise from neural stem cells before differentiating into myelinating oligodendrocytes shortly after birth. Fyn tyrosine kinase (Fyn) has been shown to play a central role during OPC differentiation, including inducing morphological changes in the cells and initiating the expression of myelin basic protein (MBP), a major structural protein required for the compaction of myelin sheaths. Recently, we have shown that signaling via the gamma chain of immunoglobulin Fc receptors (FcRgamma) induces the Fyn-MBP cascade and promotes the morphological differentiation of OPCs. The protein tyrosine phosphatases that are responsible for the positive regulation of Fyn tyrosine kinase activity during this cascade, however, remained unknown. Here we report that a protein tyrosine phosphatase, CD45, is involved in this process. Fyn co-immunoprecipitated with CD45 from differentiating wild-type OPCs in vitro, while CD45-deficient OPCs failed to differentiate. Additionally, dysmyelination was observed in CD45-deficient mice in vivo. Our findings suggest that CD45 is a key phosphatase involved in OPC differentiation and provide a preliminary explanation for the previously reported CD45 mutations observed in some multiple sclerosis (MS) patients. PMID- 15823428 TI - Attention deficits in bipolar disorder: a comparison based on the Continuous Performance Test. AB - Although attentional deficits measured by Continuous Performance Tests (CPTs) have been observed in patients with bipolar disorder, their relationship with clinical state is not well understood. The identical pairs Continuous Performance Test (CPT-IP) shows particular promise as a measure sensitive to trait abnormalities in attentional function. In this study, the CPT-IP was administered to 27 patients with bipolar disorder (22 type I, 5 type II) and 25 demographically matched healthy comparison subjects, in order to assess the presence and nature of attentional deficits as a function of mood symptoms. Results showed significantly impaired CPT performance in bipolar patients compared with healthy subjects. Patients made fewer hits (p < 0.01), were slower to respond (p < 0.007), and had poorer discrimination (p < 0.05) and bias (p < 0.006) than comparison subjects. Severity of mania and depression was not correlated with any of the CPT measures. Our findings suggest that attentional dysfunction may be a trait deficit associated with bipolar illness. However, within-subjects longitudinal studies examining fluctuations in performance over time are needed. PMID- 15823429 TI - The role of blind humans' visual cortex in auditory change detection. AB - Several studies using brain imaging have demonstrated occipital-cortex activation in blind individuals during tactile and auditory tasks, suggesting that the visual cortex deprived of its normal input has adopted a new role in information processing. So far, however, at what stages of information processing and to which perceptual sub-processes this applies remains unclear. We determined the auditory functions of this cortical region in early-blind humans by means of functional magnetic resonance imaging. We found that these areas were not activated by the mere presence of sound, but were involved in the attentive processing of changes in the auditory environment, which is important in detecting potentially dangerous or other important events in the surroundings, for example. PMID- 15823430 TI - The uptake and transport behavior of berberine in Coptidis Rhizoma extract through rat primary cultured cortical neurons. AB - The objective of this study was to investigate whether berberine in Coptidis Rhizoma extract could be transported in primary cultured cerebral neurons and to study the uptake and transport behavior of berberine through the neurons. The present results showed that the uptake of berberine by the neurons increased linearly with increasing concentration of berberine in Coptidis Rhizoma extract over the range of 0.3822-38.22 microg/mL. Berberine could be transported into the neurons in a concentration-dependent and time-dependent manner when the neurons were fed with the incubation medium containing Coptidis Rhizoma extract. There was a positive correlation between the concentration of berberine in the medium and in the cell, which means that uptake of berberine increased in the cell along with higher dose of the extract. But there was a negative correlation tendency between the concentration of berberine in the medium and the penetration ratio. It is suggested that berberine in Coptidis Rhizoma extract was transported into the neurons at a variable rate. PMID- 15823431 TI - Rhodopsin promoter-EGFP fusion transgene expression in photoreceptor neurons of retina and pineal complex in mice. AB - Light detection in vertebrate eyes is mediated through retinal photoreceptor rod and cone cells that transduce light signals into electrical responses. The differentiation and synaptogenesis of photoreceptor cells are especially important since these cells are the main targets of degeneration in retinitis pigmentosa. We produced transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of the mouse rhodopsin gene promoter. In Western blot analyses of transgenic retinal homogenates, expression of the endogenous rhodopsin gene was detected from post-natal day (P)8; however, EGFP expression in transgenic retinas was initially detected at P12, indicating delayed expression of the transgene. At P14, fluorescence microscopy showed a weak expression of EGFP in the transgenic retina. In the adult transgenic mice, the strongest EGFP expression was observed in the outer nuclear layer, and to a lesser extent in the outer plexiform layer as well as in the inner and outer segments. EGFP expression was also observed in the pineal stalk. The rhodopsin promoter-EGFP transgenic mice will be not only useful to assess rhodopsin gene promoter activity in vivo, but also for retinal transplant studies as a source of functional photoreceptor cells that are fluorescent green. PMID- 15823432 TI - Task-dependent activation latency in human visual extrastriate cortex. AB - Event related potentials (ERPs) were recorded from subjects who had to perform either an identification task or a simple detection task on moving visual stimuli. Results showed that the amplitude of the so-called visual "N1" component was larger for identification than for mere detection, replicating previous data obtained with static stimuli. However, we also found that: (i) the onset, peak and offset latencies of the visual N1 to dynamic stimuli were significantly earlier in the detection task than in the identification task, and (ii) in both conditions, the coordinates of the equivalent current dipoles best explaining the visual N1 component were consistent with those of the human motion visual area MT+/V5 in the extrastriate cortex. Altogether, these results indicate that dynamic stimuli may activate (at least partly) different pathways and processes in extrastriate cortex according to the nature of the task required on these stimuli. PMID- 15823434 TI - Epidemiology of obesity in the United States. AB - The challenges of the epidemic are not limited to concerns about bulk and weight. The disabilities caused by obesity are physiologic and psychosocial. The increased waist to hip girth is associated with increased risk of cardiovascular disease, hyperlipidemia, hypertension, and diabetes. Obesity also has been related directly to increased risk of sleep apnea, cancer, gallbladder disease, musculoskeletal disorders, severe pancreatitis, bacterial panniculitis, diverticulitis, infertility, urinary incontinence, and idiopathic intracranial hypertension. The psychosocial factors and quality of life in the obese population also have been documented. Although there is some debate, the obese have been found to be twice as likely to suffer from anxiety, impaired social interaction,and depression when compared with the nonobese population. Although advances in obesity surgery have resulted in long-term, lasting treatment of this disease and some of its comorbidities (ie, diabetes, hypertension, sleep apnea), There is a pressing need to develop a comprehensive medical and nutrition plan to reduce the prevalence of this newly identified disease state. Some draw parallels to tobacco and the morbidity and mortality associated with its use. Perhaps there are similarities in these two epidemics. Both start with education of the population as to the morbidities and mortality associated with the disease. As with tobacco, this education is especially important for youth. Without a plan of education to promote nutrition and increased physical activity, and continued research into the causes of obesity, the prevalence of obesity will continue to rise in the United States. PMID- 15823435 TI - Metabolic implications of obesity: before and after gastric bypass. AB - Obesity is recognized as an inherited disease state, and attributes of modern civilization have enhanced its phenotypic penetrance greatly. Two thirds or more of obese persons suffer comorbidities,many of which are characteristic of (dys)metabolic syndrome of insulin resistance. Unfortunately patients with body mass index over 35 infrequently can achieve or maintain weight loss adequate to resolve these metabolic (and anatomic) issues by lifestyle or pharmacologic strategies and are served better by gastric bypass, in spite of its attendant risks, both surgical and nutritional. This article evaluates the metabolic consequences of obesity and highlights which of these are amenable to correction with weight reduction achieved by gastric bypass. PMID- 15823436 TI - Metabolic complications of bariatric surgery: diagnosis and management issues. AB - Acute complications after bariatric surgery include rhabdomyolysis and immobilization hypercalcemia. The most common long-term metabolic complications following bariatric surgery are anemia and metabolic bone disease. Neuropathy, acute Wemicke's encephalopathy and vitamin A deficiency also occur. Prevention, diagnosis,and treatment of these disorders are necessary parts of lifelong care after bariatric surgery. PMID- 15823437 TI - Gastroesophageal reflux disease and obesity. AB - Logic would suggest that obesity would be a strong risk factor for causing GERD and certainly for exacerbation of GERD. Though the balance of the epidemiologic data support a relationship, true cause and effect cannot be documented. Thus, making the recommendation to lose weight to the GERD patient who is obese is reasonable. Other health reasons, however, supersede GERD as the primary impetus to lose weight. PMID- 15823438 TI - Hepatic complications of obesity. AB - Obesity increases the risk and severity of liver disease. The most common form of liver disease that occurs in obesity is NAFLD. A better understanding of the basic disease mechanisms and natural histories ofNAFLD is needed to guide management and treatment of obese patients and others with this disorder. Large, prospective, randomized, controlled treatment trials also are needed. Ideally, such studies will focus on well-defined patient subsets and monitor the impact of therapy on clinically relevant endpoints, such as liver-related morbidity and mortality. PMID- 15823439 TI - Obesity and disease of the esophagus and colon. AB - This article discusses the evidence linking obesity to the different manifestations of gastroesophageal reflux disease and colorectal cancer. PMID- 15823440 TI - Diet options of obesity: fad or famous? AB - This article highlights the different dietary approaches that have evolved and become popular over the past few years. Whether these are fads or truly represent important changes in the way peo-ple eat and that ultimately will lead to a lower rate of obesity and improved health is impossible to know. PMID- 15823441 TI - Pharmacological therapies for obesity. AB - Pharmacological therapy for obesity is in transition. Historically,there have been few effective agents, and many have been with-drawn because of unacceptable side effects. Current options include three medications approved by the FDA for the treatment of obesity: phentermine, sibutramine and orlistat. Phentermine and sibutramine suppress appetite and promote thermogenesis,and orlistat blocks fat digestion and absorption in the gut. Several drugs approved for other indications often promote weight loss, including bupropion, metformin, topiramate and zonisamide; they have been used empirically for treatment of obesity and to counter the weight-promoting effects of other medications. Expanding knowledge of the physiological mechanisms of body weight regulation has revealed new molecular targets, and more than 150 novel agents are under active development. Because weight regulation is complex, and redundant systems protect against perceived starvation, optimal treatment of obesity will likely require combinations of therapies. The accelerating emergence of new medications will facilitate the development of such effective combinations. PMID- 15823442 TI - Gastrointestinal management of the bariatric surgery patient. AB - Beyond the acute perioperative period, the most common complications of weight loss surgery relate to GI tract structure, function, and mucosal integrity. As a result, gastroenterologists have a major role in the management of patients undergoing these procedures. Optimal care of the bariatric surgical patient requires a multi-disciplinary team to address the medical complications, nutritional management, and psychological and behavioral implications of obesity. Because of their important role in preoperative assessment and postoperative management, gastroenterologists should be integral members of these multi disciplinary teams. A model of close collaboration among gastroenterologists, bariatric surgeons, and other members of the team will help optimize care of the bariatric patient and set the stage for effective development, testing, and use of the many new laparoscopic, endoscopic, intraluminal, and pharmacological GI based therapies for obesity that are under development. PMID- 15823443 TI - Surgical options for obesity. AB - Several bariatric procedures are available that have excellent long-term weight loss results and are backed by several large clinical trials. Purely restrictive procedures like VBG have fallen out of favor because of inadequate long-term weight loss. Gastric bypass and the BPD are well-studied and show significant resolution of obesity-related comorbidities. Long-term nutritional consequences are seen more commonly after malabsorptive procedures like the BPD than after hybrid malabsorptive-restrictive procedures like the gastric bypass. Because compliance and long-term nutritional follow-up are mandatory after any bariatric procedure, purely malabsorptive procedures should be reserved for super obese patients who are at risk for inadequate long-term weight loss. Furthermore, minimally invasive techniques have evolved and essentially have eliminated the high incidence of postoperative wound complications and incisional hernias frequently seen after open gastric bypass. Until the development ofa similarly successful procedure, gastric bypass will continue to be the gold standard bariatric procedure with its concurrent sustained weight loss benefits and resolution of comorbidities. PMID- 15823444 TI - Endoscopic treatments for obesity: past, present and future. AB - Obesity is recognized as a serious, chronic illness affecting all ages. The cause for obesity is multi-factorial, which makes successful management complex and challenging. Meaningful weight loss is frequently difficult to achieve, particularly when the goal is not merely to lose weight but to maintain this loss. The efficacy of future endoscopic approaches needs to be validated through well-designed controlled studies, and verification of safety is essential. Endoluminal therapies must have a sound physiologic basis for their development. The availability of specialized radionuclide imaging of gastric capacity and accommodation provide a useful tool in constructing interventions. Endotherapy is likely best suited for nonmorbid obese individuals with BMI ranging from 30 to 39 or as a budge to bariatric surgery. This specific BMI range has been targeted by the National Institutes of Health for emerging technologies. Presurgical weight loss to reduce surgical risk is another potential target group. Regardless of which endoscopic methods prevail, patients will continue to require a comprehensive, multi-modality management approach to this complex disease. PMID- 15823445 TI - Endoscopy in the bariatric surgical patient. AB - Endoscopy in the bariatric surgical patient presents a new challenge for gastroenterologists, one that will be encountered with increasing frequency as the performance of bariatric surgery increases. To ensure a safe, successful, and useful endoscopy, it is important to be familiar with the expected postsurgical anatomy and endoscopic appearance after the various bariatric operations and common complications that can arise. PMID- 15823446 TI - Lack of systemic oxidative stress during PAF challenge in mild asthma. AB - To further establish the role of oxidative stress in the pathogenesis of acute bronchial asthma, we investigated the effects of platelet-activating factor (PAF) challenge on systemic oxidant-antioxidant balance in 12 asthmatic patients (age, 25+/-3[SEM] yr; FEV1, 95+/-10% predicted), using a double blinded, controlled with Lyso-PAF (L-PAF), cross-over design. Respiratory system resistance (Rrs), arterial blood gases, peripheral blood neutrophils and oxidant-antioxidant balance, including thiobarbituric acid (TBA)-malondialdehyde (MDA) adducts, protein sulphydryls and Trolox equivalent antioxidant capacity (TEAC), were assessed at baseline and 5, 15 and 45 min after PAF and L-PAF (18 microg each) bronchoprovocation. Urinary leukotriene E4 (uLTE4) elimination was measured 120 min after challenge. Compared with baseline, as expected, PAF increased significantly Rrs and AaPO2 and decreased PaO2 and peripheral blood neutrophils along with a rebound neutrophilia and increased uLTE4. By contrast, markers of systemic oxidative stress remained unaltered throughout the study. Unlike PAF, L PAF-induced changes were negligible. We conclude that there is no systemic oxidant-antioxidant imbalance during acute bronchoconstriction induced by PAF in these patients with mild asthma. PMID- 15823447 TI - Bronchodilator response to formoterol after regular tiotropium or to tiotropium after regular formoterol in COPD patients. AB - We conducted a randomized, crossover trial with tiotropium 18 microg once daily (group A), and formoterol 12 microg twice daily (group B) over a 5-day period for each drug, with a 10-day washout, in 20 COPD patients. At the end of each period, patients inhaled both drugs separated by 180 min in alternate sequence (group A: tiotropium 18 microg+formoterol 12 microg; group B: formoterol 12 microg+tiotropium 18 microg). FEV1 and FVC were measured at baseline and after 30, 60, 120, 180, 210, 240, 300 and 360 min. FEV1 and FVC further improved after crossover with both sequences. The mean maximal change in FEV1 over baseline was 0.226 L (0.154-0.298) after tiotropium+formoterol and 0.228 L (0.165-0.291) after formoterol+tiotropium; the mean maximal change in FEV1 over pre-inhalation the second drug value was 0.081 L (0.029-0.133) after tiotropium+formoterol and 0.054 L (0.016-0.092) after formoterol+tiotropium. The mean maximal change in FVC over baseline was 0.519 L (0.361-0.676) after tiotropium+formoterol and 0.495 L (0.307 0.683) after formoterol+tiotropium; the mean maximal change in FVC over pre inhalation of the second drug value was 0.159 L (0.048-0.270) after tiotropium+formoterol and 0.175 L (0.083-0.266) after formoterol+tiotropium. The FEV1 AUCs(0-360 min) were 62.70 (45.67-79.74) after tiotropium+formoterol and 69.20 (50.84-87.57) after formoterol+tiotropium, the FEV1 AUCs(0-180 min) were 24.70 (18.19-31.21) after tiotropium+formoterol and 29.74 (21.02-38.46) after formoterol+tiotropium, whereas the FEV1 AUCs(180-360 min) were 15.70 (10.88 20.52) after tiotropium+formoterol and 11.71 (7.21-16.21) after formoterol+tiotropium. Differences between the two treatments were not statistically significant (P>0.05). The addition of second different long-acting bronchodilator to a regularly administered long-acting bronchodilator seems to be to patient's advantage. PMID- 15823448 TI - Effect of nasal continuous positive airway pressure in uncontrolled nocturnal asthmatic patients with obstructive sleep apnea syndrome. AB - The mechanisms of nocturnal asthma are intimately related to circadian rhythms, which influence inflammatory cells and mediators, hormone levels and cholinergic tone. Nocturnal airway narrowing in asthma is sometimes associated with sleep disorders, such as obstructive sleep apnea syndrome (OSAS). The aims of this study were to evaluate the association of nocturnal asthma and OSAS, and investigate the influence of continuous positive airway pressure (CPAP) therapy to improve nighttime symptoms in asthmatic patients with OSAS. Forty-three asthmatic patients who had nocturnal symptoms in spite of the optimal medical treatment according to the Global Initiative for Asthma guidelines and associated with snoring were studied. Pulmonary function tests (PFTs), asthma nighttime symptom scores, and polysomnography were performed on all patients. We treated the patients with an apnea-hypopnea index (AHI) 15 (moderate-severe OSAS) (n=16) with CPAP during 2 months. After 2 months, PFT, asthma nighttime symptom scores were reperformed. There was no significant difference in PFT values before and after CPAP treatment in OSAS patients. Asthma nighttime symptom scores were improved significantly (P<0.05) after CPAP treatment. In conclusion, in some patients with nocturnal asthma, OSAS may be responsible disease for nocturnal symptoms. In this condition, CPAP improves nocturnal symptoms without amelioration in PFT abnormalities. PMID- 15823449 TI - Cutaneous and serological responses to cat allergen in adults exposed or not to cats. AB - BACKGROUND: The relationship between pet ownership and the risk of developing respiratory allergic sensitization to pet allergens is still controversial. OBJECTIVE: To determine the degree of cutaneous immediate hypersensitivity and the levels of specific IgE and IgG4 antibodies to cat allergen in cat sensitized patients directly or indirectly exposed to this animal. METHODS: We studied 112 adolescents and adults sensitized to cat allergens (43 with and 69 without a cat at home). There were also 52 control subjects, 27 atopic non-sensitized to cat and 25 non-atopic. The degree of immediate hypersensitivity was assessed by using, in duplicate, skin prick test with four five-fold dilutions of cat hair allergen extract with the content of its major allergen Fel d 1 quantified in micrograms plus positive (10 mg/ml histamine chlorhydrate) and negative (saline solution) controls. The resulting wheal areas were analysed by means of Parallel Line Assay. A blood sample was collected from every patient and control subjects for the evaluation of serological cat specific IgE and IgG4 antibodies. RESULTS: Patients with cat at home had a lower cutaneous response than patients without this pet. The difference in the skin sensitivity was estimated in 3.4 times (P<0.01). There was no statistical difference between the levels of cat specific IgE antibodies in the two groups of patients (P=0.065). The levels of Fel d 1 specific IgG4 antibodies showed a statistically significant association with the presence of cat at home, with higher levels in patients owing cat at home than in patients without this pet (P<0.001). CONCLUSION: The results of this study demonstrate that direct cat exposure in adolescents and adults with respiratory allergy is associated with a lower cutaneous response to cat allergenic extract, assessed by SPT and compared with indirect exposure. In patients with cat at home mean levels of specific IgE are statistically comparable whereas the levels of IgG4 are higher in comparison with subjects not exposed to cats. The role of indirect exposure to cat allergens in airways sensitization also in adults is emphasized. Moreover, patients with cat at home show a cutaneous and serological sensitization to cat allergen not higher in comparison with subjects not exposed to cats. PMID- 15823450 TI - The 24 h duration of bronchodilator action of the salmeterol/fluticasone combination inhaler. AB - INTRODUCTION: The duration of bronchodilator action of the long-acting beta agonist salmeterol when administered in the evening has not been investigated. In this study we have investigated whether a single evening dose of salmeterol, administered from the combination salmeterol/fluticasone (SFC) Accuhaler significantly attenuates the circadian rhythm in airway tone over 24 h. METHODS: Eighteen subjects with mild to moderate asthma (mean FEV1 84% predicted) participated in a double-blind, double dummy, placebo controlled, cross-over study. Subjects inhaled, in random order, placebo, salbutamol (200 microg) or SFC (50/100 microg) administered in the evening (2000 h) on three separate occasions. Lung function measurements including FEV1, specific airways conductance (sGaw) and maximum expiratory flow at 25-75% of vital capacity (MEF(25-75%)) were assessed at baseline, at 1 h and subsequently every 4 h post-dose for 24 h. RESULTS: Compared with placebo, SFC significantly improved the three measures of airways function throughout the 24 h period, with a difference in FEV1 at 24 h of 0.24 l (0.00-0.47 l). SFC abolished the biphasic pattern of the circadian rhythm in airway tone. In contrast, salbutamol had a significant bronchodilator action of 4-8 h, depending on the lung function parameter measured. CONCLUSION: The single evening administration of SFC via the Accuhaler resulted in a duration of bronchodilation of at least 24 h, with the abolition of the accentuated biphasic circadian variation in airway tone observed in asthma. PMID- 15823451 TI - Measurement properties and interpretation of three shortened versions of the asthma control questionnaire. AB - The Asthma Control Questionnaire (ACQ) measures the adequacy of asthma treatment as identified by international guidelines. It consists of seven items (5 x symptoms, rescue bronchodilator use and FEV1% of predicted normal). A validation study suggested that in clinical studies measurement of FEV1 and bronchodilator use may not be needed but this has never formally been tested in a clinical trial. The aims of this analysis were (1) to examine the measurement properties of three shortened versions of the ACQ (symptoms alone, symptoms plus FEV1 and symptoms plus short-acting beta2-agonist) and (2) to determine whether using the shortened versions would alter the results of a clinical trial. In the randomised trial, 552 adults completed the ACQ at baseline and after 13 and 26 weeks of treatment. The analysis showed that the measurement properties of all four versions of the ACQ are very similar. Agreement between the original ACQ and the reduced versions was high (intraclass correlation coefficients: 0.94-0.99). Mean differences between the ACQ and the shortened versions were less than 0.04 (on the 7-point scale). Clinical trial results using the four versions were almost identical with the mean treatment difference ranging from -0.09 (P=0.17), to 0.13 (P=0.07). For interpretability, the minimal important difference for all four versions was close to 0.5. In conclusion, these three shortened versions of the ACQ can be used in large clinical trials without loss of validity or change in interpretation. PMID- 15823452 TI - Analysis of insulin like growth factor 1 and insulin like growth factor binding protein 3 levels in bronchoalveolar lavage fluid and serum of patients with lung cancer. AB - OBJECTIVE: Insulin like growth factor 1 (IGF-1) is recognized as a potent mitogen for many cancer cell lines and there is good evidence that lung cancer cells produce both IGF-1 and insulin like growth factor binding protein 3 (IGFBP-3). The aim of this study was to investigate the clinical significance of IGF-1 and IGFBP-3 levels in serum and in bronchoalveolar lavage (BAL) fluid by comparing lung cancer patients with healthy controls. METHODS: BAL fluid and serum samples were obtained from 24 lung cancer patients and 12 healthy controls, and were analyzed for IGF-1 and IGFBP-3 levels by a two site immunoradiometric assay. The recovered BAL fluid was standardized by albumin to remove the variable of dilution and the data was expressed in epithelial lining fluid (ELF). RESULTS: Serum IGF-1 and IGFBP-3 levels were lower in lung cancer patients, but the difference between the groups did not reach a statistical significance. IGF 1/IGFBP-3 ratio in ELF was significantly lower in lung cancer patients (P=0.035). Mean IGF-1 level in ELF was determined to be significantly lower in patients with distant metastasis (P=0.04). Serum IGF-1/IGFBP-3 ratio was found to be significantly lower in patients with distant (P=0.04) and nodal metastasis (P=0.03). Tumor stage was negatively correlated with IGF-1 level in ELF (P=0.05, r=-0.4) and serum IGF-1/IGFBP-3 ratio (P=0.04, r=-0.4). CONCLUSION: IGF-1 and IGFBP-3 levels both in serum and ELF might serve a clinical significance in patients with lung cancer. However, further studies comprising more cases are needed to investigate the clinical significance of IGF-1 and IGFBP-3 in lung cancer. PMID- 15823453 TI - Rapid onset of tolerance to beta-agonist bronchodilation. AB - INTRODUCTION: Regular use of beta-agonists leads to tolerance to their bronchodilator effects. This is easily demonstrated if dilation is tested following methacholine challenge. It is not known how quickly tolerance develops or how long it lasts after stopping beta-agonist therapy. METHODS: Ten subjects with stable asthma were studied. Following 2 weeks without beta-agonists, methacholine was inhaled to induce a 20% reduction in FEV1. The response to inhaled salbutamol (100, 100, 200 microg at 5-min intervals) was then measured. This procedure was repeated 24 h after one dose and 24 h after 3, 7 and 14 days of inhaled formoterol 12 microg twice daily, and 3 and 5 days after formoterol was discontinued. Unscheduled use of beta-agonists was not permitted. RESULTS: Bronchodilator tolerance, assessed by a reduction of the area under the salbutamol dose-response curve, occurred after 1 dose of formoterol (28% reduction, 95% CI 12, 45%), increased up to 1 week and plateaued between 1 and 2 weeks (58% reduction, 95% CI 38, 78%). Three days after stopping formoterol, the response to salbutamol was similar to baseline (12% reduction, 95% CI -9, 33%). The first dose of formoterol provided significant bronchoprotection to methacholine (1.6 doubling doses, P=0.007). This diminished with regular treatment and by 2 weeks the PD20 methacholine was not significantly different to baseline. CONCLUSIONS: Bronchodilator tolerance occurs after a single dose and reaches a maximum after 1 week of regular formoterol. Sensitivity recovers 3 days after stopping treatment. PMID- 15823454 TI - Changes in sputum T-lymphocyte subpopulations at the onset of severe exacerbations of chronic obstructive pulmonary disease. AB - CD8+ve T-cell responses play a primary role in chronic obstructive pulmonary disease (COPD), but there is little information regarding COPD exacerbations. Sputum induction is a relatively non-invasive and safe method to study airway inflammation. The aim of the study was to investigate changes in airway T lymphocyte subpopulations at the onset of severe COPD exacerbations via analysis of sputum. Induced sputum samples were collected from 12 COPD patients aged (mean+/-sd) 69+/-7 years, ex-smokers (68+/-23 pack-years), mean FEV1 (%predicted) 40+/-14 at the onset of an acute severe exacerbation requiring hospital admission and 16 weeks after remission of the exacerbation. Inflammatory cells and T lymphocyte subpopulations (CD4, CD8, Tc1, Tc2) were measured using chemical and double immunocytochemical methods. Increased percentages of sputum neutrophils (P=0.002) and decreased CD4/CD8 and CD8-IFNgamma/CD8-IL4+ve (Tc1/Tc2) cell ratios (P=0.03, P=0.02, respectively) were found at the onset of exacerbation compared to stable state. We conclude that a CD8+ve type-2-mediated immune response is induced at the onset of severe COPD exacerbation. PMID- 15823455 TI - Effects of antioxidants and NO on TNF-alpha-induced adhesion molecule expression in human pulmonary microvascular endothelial cells. AB - Pro-inflammatory cytokines initiate the vascular inflammatory response via upregulation of adhesion molecules on the endothelium. Recent observations suggest that reactive oxygen intermediates may play a pivotal role in TNF-alpha signaling and upregulate gene expression. We therefore evaluated the effects of pyrrolidine dithiocarbamate (PDTC; 0.1 mM) and spermine NONOate (Sper-NO; 1 mM) on adhesion molecule expression and nuclear factor kappa B (NF-kappaB) activation induced by TNF-alpha (10 ng/ml) in cultured human pulmonary microvascular endothelial cells (PMVEC). Treatment of cells with TNF-alpha for 4 h significantly induced the surface expression of E-selectin and ICAM-1. Treatment with TNF-alpha for 8 h significantly induced the surface expression of E selectin, ICAM-1 and VCAM-1. The upregulation of these adhesion molecules was suppressed significantly by pretreatment with PDTC or Sper-NO for 1 h. 8-Bromo cyclic GMP (1 mM) had no such effect, suggesting that the NO donor's effect was non-cGMP-dependent. The mRNA expression of E-selectin, ICAM-1 and VCAM-1, and activation of NF-kappaB induced by TNF-alpha for 2 h were decreased significantly by the above two pretreatments. N-acetylcysteine (10 mM) and S-nitroso-N acetylpenicillamine (1 mM) had little inhibitory effects on the cell surface and mRNA expression of these adhesion molecules stimulated by TNF-alpha. Treatment with TNF-alpha for 4 h enhanced HL-60 leukocyte adhesion to human PMVEC, the effect of which was inhibited significantly by pretreatment with PDTC or Sper-NO. These findings indicate that both cell surface and mRNA expression of adhesion molecules in human PMVEC induced by TNF-alpha are inhibited significantly by pretreatment with PDTC or Sper-NO, possibly in part through blocking the activation of NF-kappaB. Although our in vitro results cannot be directly extrapolated to the in vivo situation, they suggest a potential therapeutic approach for intervention in cytokine-mediated inflammatory processes in the human lung. PMID- 15823456 TI - Revisiting the accuracy of peak flow meters: a double-blind study using formal methods of agreement. AB - BACKGROUND: There is widespread use of peak flow meters in both hospitals and general practice. Previous studies to assess peak flow meter accuracy have shown significant differences in the values obtained from different meters. However, many of these studies did not use human subjects for peak flow measurements and did not compare meters of varying usage. In this study human subjects have been used with meters of varying usage. METHODS: Participants were tested using two new (meters A and C) and one old peak flow meter (meter B) in random order. The study was double-blinded. Participants were recruited from the university campus. RESULTS: Four hundred and nine individuals participated. The difference between peak flow means of A and B was -9.93 l/min (95% CI: -12.37 to -7.48, P<0.0001). The difference between peak flow means of B and C was 20.08 l/min (95% CI: 17.85 22.29, P<0.0001). The difference between peak flow means of A and C was 10.15 l/min (95% CI: 7.68-12.61, P<0.0001). CONCLUSION: There was a significant difference between the values obtained from the new and old peak flow meters and also between the two new peak flow meters. We conclude that there is need for caution in interchangeably using flow meters in clinical practice. PMID- 15823457 TI - Successful steroid withdrawal in lung transplant recipients: result of a pilot study. AB - OBJECTIVE: Corticosteroids play a key role in immunosuppression after transplantation. However, because chronic steroid treatment may cause significant morbidity and mortality, steroid-free immunosuppression remains a desirable goal. To the best of our knowledge, there are no reports on successful steroid withdrawal (SW) in lung transplant recipients. METHODS: The study group included 35 patients who underwent heart-lung, double-lung or single-lung transplantation. Criteria for initiation of SW were stable pulmonary function tests and absence of clinical or bronchoscopic evidence of acute or chronic rejection in the last 6 months. Pulmonary function, blood pressure and metabolic parameters were compared between the patients who underwent SW and those who did not. RESULTS: Eight patients (23%) underwent SW. Median follow-up was 19 months (range 11-23 months). Compared to the non-withdrawal group, the withdrawal group was older (60+/-6 vs. 52+/-13 years, P=0.01, r=0.49), had higher rates of emphysema (88% vs. 18%, P=0.01) and use of a cyclosporine-based regimen (62% vs. 26%, P=0.0001), and had longer time from transplantation to the withdrawal attempt (70+/-13 vs. 29+/-26 months, P=0.0002). The SW group showed no adverse effects in graft function and no deterioration on pulmonary function tests. SW had a beneficial metabolic effect, with a decrease in mean cholesterol level from 229+/-45 to 194+/-25 mg/dl (P=0.02) and no significant change in weight, systolic blood pressure or glucose level. In the non-withdrawal group, mean cholesterol levels increased from 175+/ 34 to 209+/-57 mg/dl (P=0.0005), weight increased from 72+/-15 to 80+/-14 kg (P=0.0001), and systolic blood pressure increased from 125+/-15 to 139+/-16 mmHg (P=0.001); glucose levels did not change. There was a significant correlation between total cholesterol level and weight in both groups (P=0.0006, r=-0.56 and P=0.01, r=-0.46, respectively). CONCLUSIONS: Late SW is safe in stable patients after lung transplantation. There was no evidence of rejection or a deterioration in pulmonary function. Lipid profile improvement and blood pressure stabilization accompanied the termination of steroid therapy. PMID- 15823458 TI - Brazilian version of airways questionnaire 20: a reproducibility study and correlations in patients with COPD. AB - This study describes the correlations and reproducibility of AQ20, a simple health status questionnaire with 20 questions, which was designed to be useful especially in time sparing situations. A formal language validation process was done, in order to validate the AQ20 before studying its reproducibility. Thirty stable COPD patients answered the final version twice within 15 days. To test the reproducibility of AQ20, the interclass correlation coefficient and Bland-Altman display were used. Results were correlated with FEV1, SpO2, BMI, Mahler BDI, and the Saint George Respiratory Questionnaire (SGRQ). Twenty-five patients (83.3%) were male, with a mean age of 68.6 years. The mean predicted FEV1 (%) was 56.8%. The interclass correlation ratio for the total score was alpha=0.90 for the intraobserver variability and alpha=0.93 for the interobserver variability. The correlation with total SGRQ score was 0.76, with P<0.001. The mean application time for AQ20 was 4 min and 6 s, and the score calculation time, was 8 s. It can be concluded that AQ20 is reproducible, with an excellent correlation with SGRQ total score, and also having the advantage of taking just a few minutes to be applied and to have its score calculated. PMID- 15823459 TI - Accuracy of pulmonary function tests in predicted exercise capacity in COPD patients. AB - PURPOSE: The purpose of this study was to examine exercise tolerance in patients with COPD from measurements of resting pulmonary function parameters. METHODS: A total of 57 COPD patients were administered the pulmonary function test (PFT) and cardiopulmonary exercise test. The results were analyzed and essentially linear relationships emerged when each subject's VO2 peak was plotted against his individual PFT parameters. Those significant contributors were then introduced in a stepwise multiple regression analysis to determine the best predictor of the VO2 peak. RESULTS: Stepwise multiple regressions in variables revealed that peak oxygen consumption (VO2 peak) was predicted best by the following equation: VO2 peak=(maximum voluntary ventilation x 0.024)+(forced mid-expiratory flow x 0.47)+(body surface area x 0.988)-0.913 (r=0.90; r2=0.81 SE=0.29 L/min). CONCLUSION: We conclude that exercise capacity was predicted from measurements of resting pulmonary function parameters with excellent accuracy in the COPD patient. PMID- 15823460 TI - Rounded atelectasis of the lung. AB - Rounded atelectasis of the lung (RA) is a lesion well described in the medical literature, yet often very difficult to diagnose. In recent years, the widespread use of high-resolution imaging modalities employed in the struggle against cancer, coinciding with the peak of the asbestos epidemic, have boosted the detection frequency of RA. However, its differential diagnosis still poses a challenge to the pulmonary specialist and the radiologist, as little is known about its pathogenesis. Furthermore, the multifactorial etiology of RA and its occasional coexistence with lung cancer make the task of confidently ruling out malignancy sometimes daunting. This article attempts to provide an update on RA's etiology, radiological evaluation, clinical management, and prognosis based on recent advances in broadly available diagnostic modalities and minimally invasive interventional procedures. An exemplary case of post-tuberculous RA is illustrated, as RA often presents as an unusual finding of a fairly common disease. PMID- 15823461 TI - Both generic and disease specific health-related quality of life are deteriorated in patients with underweight COPD. AB - This study evaluated the effects of body weight on both generic and disease specific health-related quality of life (HRQoL) of patients with COPD. A total of 83 patients with stable COPD were enrolled (mean age: 74.6 yr, mean FEV1: 1.29 L). Patients were divided into two groups according to body mass index (BMI) (UW group: BMI < or = 20, NW group: 20 < BMI < or = 26). The degree of dyspnea and both disease-specific and generic HRQoL were compared between the two groups. An oxygen cost diagram (OCD) was used to assess the degree of dyspnea and St. George's Respiratory Questionnaire (SGRQ) and Medical Outcomes Study Short Form 36-item Questionnaire (SF-36) were used for HRQoL evaluation. The OCD was significantly lower in the UW group. Compared with the NW group, the UW group showed significant deterioration in the total score and three subscales of SGRQ. SF-36 also showed significantly worse scores for the parameters of physical functioning, role emotional, bodily pain, and general health. The results of stepwise multiple regression analysis showed that OCD, FEV1, %pred. BMI were independent variables in the total score on SGRQ. The results of stepwise multiple regression analysis also showed that OCD was an independent variable for four of eight components of SF-36, while BMI was three of eight components of SF 36. In conclusion, low body weight in patients with COPD is related to a worsening of dyspnea and deterioration of both generic and disease-specific HRQoL. The present results also indicate that nutritional intervention may be important for improving dyspnea and HRQoL in patients with COPD. PMID- 15823462 TI - Comparison of exhaled nitric oxide analysers. AB - Currently no published data are available concerning the comparability of different types of NO analysers, making inter-laboratory comparisons difficult. In two sets of experiments we compared 4 and 5 NO analysers, respectively, from 3 different manufacturers using different calibration regimes: calibration with (1) a separate recommended calibration gas for each analyser, (2) a single low concentration for all (394 ppb), and (3) a single high concentration (12.8 ppm). We measured three subjects with known low (L), moderate (M) and high (H) bronchial exhaled nitric oxide concentrations as well as standard gases (SG). In the first set of experiments, calibration regime 1 resulted in the largest differences between analysers (coefficient of variation (CV) for L, M, H, SG: 0.42, 0.22, 0.20, 0.14). The lowest CV between analysers was observed after calibration 2 (0.34, 0.19, 0.12, 0.02). Very similar results were obtained in the second set of comparisons. Thus, differences between analysers existed, but were mainly due to differences in recommended calibration gases/procedures. Only a small part was explainable by deviations from target flow. These differences need to be taken into account when comparing data between laboratories or replacing the calibration gas of an analyser, as well as for the establishment and interpretation of normal values. PMID- 15823463 TI - Determinants of health-related quality of life in patients with pulmonary emphysema. AB - QUESTION OF THE STUDY: The aim of our study was to examine the relationship between health-related quality of life (HRQL), lung function parameters and intensity of dyspnea in order to determinate what variables influence the HRQL in patients with pulmonary emphysema. PATIENTS AND METHODS: Forty (mean aged 66+/-9) consecutive male patients with pulmonary emphysema were evaluated. All patients underwent spirometry, measurement of lung volumes, inspiratory and expiratory respiratory pressure (MIP and MEP), measurement of corrected carbon monoxide diffusing capacity (KCO), and 6-min walking test (6MWT). The scale of dyspnea by the baseline dyspnea index (BDI) and British Medical Council Research (MCR), and the quality of life by Chronic Respiratory Question Disease (CRQD) were also assessed. RESULTS: FEV1, FVC, 6MWT, KCO, MEP, MIP, BDI and MCR were correlated with dimensions of CRQD. Factor analysis reduced these variables to four factors, which accounted for 86.2% of the total variance: (1) airway obstruction, (2) dyspnea, (3) capacity of effort, (4) maximum static respiratory pressure. In the multiple regression model BDI and MEP explained the 64% of the total variance of CRQD. CONCLUSIONS: Quality of life measured by CRQD in patients with emphysema is predominantly determined by dyspnea, and in minor degree by expiratory muscle strength. Our results underscore the usefulness of dyspnea scales and MEP in the evaluation of HRQL in patients with emphysema. PMID- 15823464 TI - Bronchoscopic intralesional injection of triamcinolone acetonide treated against bronchial obstruction caused by peanut aspiration. AB - We encountered a case of obstructive pneumonia 1 month after peanut aspiration. Bronchoscopic examination revealed obstruction of the right basal bronchus with immature granulation tissue due to the peanut aspiration. Despite antibiotic treatment, the obstruction had not been resolved. To relieve the bronchial obstruction, bronchoscopic intralesional injection of triamcinolone acetonide was performed without any complications. Seven days after injection, the bronchial obstruction had resolve. In conclusion, intralesional injection of triamcinolone acetonide may play a role in one of the non-surgical repairs against obstruction due to foreign body aspiration. PMID- 15823465 TI - Delayed response to anti-tuberculosis treatment in a patient on infliximab. AB - The introduction of infliximab, an anti-TNF-alpha agent, in the treatment of rheumatic diseases has offered important therapeutic advances in recent years. The main adverse effect from the usage of this drug is susceptibility to infections, mainly reactivation of latent tuberculosis. We present a 23 year-old male with ankylosing spondylitis, who developed endobronchial and widespread pulmonary tuberculosis, 2 years after initiation of treatment with infliximab. The patient had already been treated for a positive PPD skin test with a 9-month prophylactic course of isoniazid. He was treated with a five drug anti tuberculosis scheme but he showed an extremely slow therapeutic response with daily high fever, even 4 months after initiation of treatment. Seven months after beginning anti-tuberculosis therapy, bronchoscopy still revealed necrotic and inflammatory tissue at the site of the original lesions. This unusual clinical course of tuberculosis infection was attributed to immunosuppression due to the long-lasting anti-TNF-alpha action of infliximab. PMID- 15823466 TI - Recovery time following bronchial challenge. PMID- 15823467 TI - Some current developments in orthopaedic biomechanics. PMID- 15823468 TI - Strain rate influences periosteal adaptation in mature bone. AB - Mechanical forces influence bone form and function. Although the adaptive capabilities of bone are well known, the nuances of the mechanical stimuli regulating adaptation remain elusive. Recently, it was suggested that strain rate influences bone adaptation, and impact exercises with high strain rates during growth may be more osteogenic than low impact aerobic exercises. Building on those findings, we hypothesized that higher rates of mechanical loading would evoke greater adaptive responses than lower rates of loading in mature bone. To test that hypothesis, skeletally mature (16 weeks) female C57BL/6 mice underwent non-invasive exogenous cantilever bending of the right tibia with a 1 Hz trapezoidal waveform for 60 s, 5 days per week, for 4 weeks. Loading was calibrated (strain gauge) to induce peak magnitudes of 1000 microepsilon on the lateral tibial middiaphysis. Mice were randomly assigned to three groups based on strain rate of the applied load: low (0.004 s(-1); n = 14), medium (0.020 s(-1); n = 15), and high (0.100 s(-1); n = 14). Calcein injections (i.p., 10 mg kg(-1)) permitted histomorphometric analyses of bone formation. Loading significantly enhanced periosteal mineral apposition rate (MAR), mineralizing surface (MS), and bone formation rate (BFR BS(-1)) in all three strain rate groups, relative to control tibiae. Furthermore, a graded dose-response relation was observed between the applied strain rate and periosteal BFR BS(-1). These increases in MAR, MS, and BFR BS(-1) were not seen on the endosteal surface. Endosteal adaptation was not statistically different between loaded and control tibiae in most endosteal indices of bone adaptation. Moreover, endosteal adaptation did not increase with strain rate. Understanding the nature of the stimuli to which bone cells respond to may underpin the development of non-pharmacological treatments devised to enhance bone mass. PMID- 15823469 TI - Low amplitude, high frequency strains imposed by electrically stimulated skeletal muscle retards the development of osteopenia in the tibiae of hindlimb suspended rats. AB - The purpose of this study was to determine the extent to which high frequency, low amplitude skeletal muscle contractions, induced using electrical stimulation, could prevent or retard disuse osteopenia. Ten minutes of 30 Hz electrical stimulation was delivered, 5 days a week, during a 4 week rat-hindlimb suspension protocol. Each pulse generated a peak compressive dynamic strain on the tibia of approximately 200 microepsilon. We hypothesized that the electrical stimulation protocol would significantly reduce the loss of tibial bone mineral density compared to the contralateral control tibia that did not receive electrical stimulation. Compared to the contralateral control limb, the tibia of the stimulated limb had significantly higher bone mineral density and enhanced newly formed bone in the tibial diaphysis. The diaphysis, specifically the posterior bone cortex, of the tibia of the limb receiving the stimulation also demonstrated substantially larger mineral-binding fluorochrome biomarker within the osteocyte lacunae and canalicular volumes. Although the protocol did not prevent disuse osteopenia the evidence suggests that it was effective at reducing the extent of the osteopenia. One possibility for this outcome may be the insensitivity of bone to static, compared to dynamic compressive loads. In the present study there was a considerable static component to the compressive loads that accounted for a large component of the peak load generated by the stimulated skeletal muscle. Nevertheless, the results provide impetus for further development of the methods by which muscle contraction-induced loading of bone can be clinically exploited. PMID- 15823470 TI - The influence of mechanical properties of subchondral plate, femoral head and neck on dynamic stress distribution of the articular cartilage. AB - A few finite element models have addressed the dynamic juxtaarticular stress transmission but none focused the investigation on the combined influence of the individual moduli of the underlying bones, including the subchondral plate, the femoral head and the femoral neck of the proximal femur. A finite element model including the acetabulum and the proximal femur was analyzed with dynamic loadings to study the effects of mechanical property changes in the underlying bones of the proximal femur on the stress distribution in the cartilage at the hip joint. We found the stress distribution was most sensitive to the subchondral plate stiffening, while the overall stiffening of the underlying bones had mild effect on the shear stress on the cartilage surface (or at the subchondral bone/cartilage interface) and on the strain energy density in the cartilage. Our results indicate that the subchondral plate plays a predominant mechanical role in the initial degeneration of the cartilage. The results may offer a mechanical explanation as to why the cartilage failure is common in patients with osteoarthritis but rare in patients with osteoporosis. PMID- 15823471 TI - Spatial and temporal regulation of cancellous bone structure: characterization of a rate equation of trabecular surface remodeling. AB - A computer simulation of trabecular surface remodeling was carried out to investigate the spatial and temporal regulation of the cancellous bone structure caused by bone cellular activities responding to a local mechanical environment. In the remodeling simulation, the rate of trabecular surface movement was directly related to stress at the trabecular level. Two model parameters, the threshold value of the lazy zone and the sensing distance of the mechanical environment, were introduced into the remodeling rate equation to express the sensitivity of bone cells to mechanical stimuli. A rectangular cancellous bone model under simple and nonuniform compressive loads was constructed using pixel finite elements. A simulation result revealed that the trabecular structure underwent a temporal and spatial change depending on the loading condition. It was found that the threshold value of the lazy zone regulates the rate of structural changes in time, and that sensing distance regulates the spatial distribution of the trabecular structure. The results demonstrate the possibility that the spatial and temporal regulation of the trabecular structure is determined by the sensitivities of bone cells to mechanical stimuli. PMID- 15823472 TI - Total flavones of Hippophae rhamnoides promotes early restoration of ultimate stress of healing patellar tendon in a rat model. AB - Traditional Chinese herbal medicine has long been used for treatment of tendon injuries. Comparing to the modern way of treatments, Traditional Chinese medicine also stresses on strategies to promote the inherent healing capacity of tendons. Hippophae rhamnoides, known as Shaji, is one of Chinese herbal drugs that are traditionally used to promote tendon and ligament injuries. The total flavones of H. rhamnoides (TFH), with major constituents including quercetin, isorhamnetin and kaempferol, have been demonstrated with most of the bioactive properties of Shaji. In the present study, we evaluated the potential effect of TFH in the restoration of ultimate stress of healing patellar tendon in a well-established gap wound model in rats. A 0.1 mg TFH was injected to wound 1 day after the injury, and the ultimate stress of the healing tendon was measured at day 14 post injury. The results showed that the ultimate stress of the healing tendon was significantly promoted by injection of TFH, increasing from 30 to 50% as compared to saline control. Excessive fibrotic response was not found in TFH-treated animals, but an enhanced collagen deposition and a better fibre alignment were observed. The results suggest that TFH may improve the ultimate stress of healing tendons at early stages, which implies possible earlier rehabilitation programme and better recovery. PMID- 15823473 TI - Activity-dependence of the "safe zone" for impingement versus dislocation avoidance. AB - Presently, the basis for optimal cup positioning to minimize the likelihood of dislocation rests upon subjective clinical impressions. To help elucidate optimal cup positioning more objectively, and to clarify the distinction between impingement avoidance and dislocation avoidance, kinematic and kinetic inputs for seven dislocation-prone activities were applied to finite element models of a contemporary 22-mm modular total hip reconstruction. Twenty-five cup placement positions (combinations of 30, 40, 50, 60, and 70 degrees of abduction with 0, 10, 20, 30, and 40 degrees of anteversion) were chosen to include the conventional 'safe zone' of 30-50 degrees of tilt and 5-25 degrees of anteversion. Activities studied were: rising from a low seat (toilet) and from a normal height chair, leg-crossing while seated, tying a shoe from a seated position, bending at the hip from an erect stance to retrieve an object on the floor (stooping), a standing pivot maneuver, and rolling over in bed. Neck-on-cup impingement occurred during one or more of the dislocation-prone activities at all 25 cup positions. Of the 175 combinations of cup position and kinetic challenge, dislocation and impingement events both occurred for 51 situations, while impingement occurred in 45 instances without dislocation, and dislocation occurred in 10 instances without impingement. Neither dislocation nor impingement was observed in the 69 other combinations of cup position and loading challenge. Kappa statistics showed dislocation and impingement, as outcome measures of activity-dependent challenges, exhibit little more than chance agreement. Therefore, the use of impingement range of motion (ROM) as a predictor of a given cup position's vulnerability to dislocation should be viewed cautiously. PMID- 15823474 TI - Contact stresses in the knee joint in deep flexion. AB - The contact stresses in the knee that arise from activities involving deep flexion have not been given due consideration in view of social and cultural practice amongst many Asians that frequently cause the engagement of these activities. Excessively large stresses (>25 MPa) can cause cartilage damage and may be the precursor to the development of degenerative disease of the joint. In this study, forces in the knee derived from previous studies of human walking and squatting were applied to five cadaver knees that underwent quasi-static mechanical testing. This was conducted using a materials-testing machine and a custom-made apparatus that allowed secure and consistent loading of the knee specimen in flexion beyond 120 degrees. A thin-film electronic pressure transducer was inserted into the cadaver tibiofemoral joint space to measure force and area. Throughout the various positions simulating specific phases of walking, it was found that stresses peaked to 14 MPa (standard deviation was 2.5 MPa). In deep flexion, the peak stresses were significantly larger by over 80%, reaching the damage limits of cartilage. The results from this biomechanical study suggest that the adequacy of articular cartilage to support loads in the knee joint during deep flexion may be questionable. PMID- 15823475 TI - The kinematic importance of radial neck length in radial head replacement. AB - Comminuted radial head fractures can be treated with a radial head implant. The effects of lengthening (2.5 mm, 5 mm) and shortening (2.5 mm, 5 mm) of the radial neck, were compared to the nominal length in six human upper extremity cadavers. Total varus-valgus laxity and ulnar rotation were recorded. We hypothesized that restoring the exact length of the radius is important to maintain normal kinematics in the elbow joint. Lengthening or shortening of more than 2.5 mm significantly changed elbow kinematics. Lengthening caused a significant decrease (p < 0.001) in varus-valgus laxity, with the ulna tracking in varus and external rotation. Shortening caused a significant increase in varus-valgus laxity (p < 0.001) and ulnar rotation (p < 0.001), with the ulna tracking in valgus and internal rotation. Our study suggests that a restoration of radial length is important and that axial understuffing or overstuffing the radiohumeral joint by 2.5 mm or more, will alter elbow kinematics. PMID- 15823476 TI - Development and calibration of a load sensing cervical distractor capable of withstanding autoclave sterilization. AB - In surgery of the cervical spine, a Caspar pin distractor is often used to apply a tensile load to the spine in order to open up the disc space. This is often done in order to place a graft or other interbody fusion device in the spine. Ideally a tight interference fit is achieved. If the spine is over distracted, allowing for a large graft, there is an increased risk of subsidence into the endplate. If there is too little distraction, there is an increased risk of graft dislodgement or pseudoarthrosis. Generally, graft height is selected from preoperative measurements and observed distraction without knowing the intraoperative compressive load. This device was designed to give the surgeon an assessment of this applied load. Instrumentation of the device involved the application of strain gauges and the selection of materials that would survive standard autoclave sterilization. The device was calibrated, sterilized and once again calibrated to demonstrate its suitability for surgical use. Results demonstrate excellent linearity in the calibration, and no difference was detected in the pre- and post-sterilization calibrations. PMID- 15823477 TI - Defining the Parkinson's disease phenotype: initial symptoms and baseline characteristics in a clinical cohort. AB - OBJECTIVES: Examine the demographic and historical characteristics of a large Parkinson's disease (PD) clinical cohort and determine the pattern and relationship between initial symptoms of PD and physical examination ratings at the initial subspecialty clinic visit. METHODS: A clinical series of 1244 consecutive individuals diagnosed with PD. Baseline characteristics were examined. RESULTS: The overall sample was predominantly right-handed (92%) male (67%), with a mean age of 70 years and symptomatic disease duration of about 7 years. About 25% of the sample reported a positive family history of a neurodegenerative disorder. Tremor (47%) and bradykinesia (29%) were the two most common initial symptoms of PD and occurred primarily in the upper extremities (68%). Tremor, bradykinesia, and postural instability showed a significant correspondence between the initial self-reported symptom and clinical signs at the first exam p < 0.05. However, rigidity did not show a significant correspondence. CONCLUSION: The demographic and clinical characteristics of the sample are thought to be phenotypically representative of 'typical' PD and were comparable to other clinic- and population-based cohorts. Initial symptoms reported by patients, including type and body location correlate tightly with initial examination findings for all cardinal symptoms of parkinsonism except rigidity. This corroboration may substantiate inclusion of such data in studies of longitudinal progression in PD. PMID- 15823478 TI - Sporadic SCA8 mutation resembling corticobasal degeneration. AB - Spinocerebellar ataxia type 8 (SCA8) is caused by the expansion of CTA/CTG triplet repeats on 13q21. Cases can be familial or sporadic. The clinical findings include cerebellar ataxia with upper motor neuron dysfunction, dysphagia, peripheral sensory disturbances, or cognitive and psychiatric impairments, indicating phenotypic variability in SCA8. We report on a patient with rapidly progressive parkinsonism-plus syndrome resembling corticobasal degeneration and triplet expansions in the SCA8 locus. The relationship between clinical phenotype and triplet expansions in the SCA8 locus requires further study. PMID- 15823479 TI - Dyskinesias predict the onset of motor response fluctuations in patients with Parkinson's disease on L-dopa monotherapy. AB - The aim of the study was to investigate the relationship between dyskinesias and motor fluctuations in patients with Parkinson's disease on l-dopa monotherapy. We identified 116 patients on l-dopa monotherapy treated between 1965 and 1992 and followed until death. Dyskinesias occurred in 102 patients. Of these, 48 only developed dyskinesias while 54 had both dyskinesias and motor fluctuations. Among patients with both complications, 49 developed dyskinesias before fluctuations, and only five had dyskinesias after the onset of fluctuations. Our findings suggest that dyskinesias predict the onset of motor fluctuations, and may share a common pathophysiological mechanism. PMID- 15823480 TI - Long-term effects of bilateral subthalamic nucleus stimulation in advanced Parkinson disease: a four year follow-up study. AB - In this study we aimed to investigate the effects of bilateral STN HFS in patients with advanced Parkinson disease (PD) at long-term, with a minimum follow up of 4 years. Twenty patients (15 men, five women) were included, with a mean age of 60.9+/-8.1 years. Surgery was performed under local anesthesia. The target was defined on computerized tomography (CT). At 3 months, significant improvements were found on the total Unified Parkinson disease rating scale (UPDRS) III (motor) score, in the medication. off (from 42.3+/-9.3 to 19.5+/ 6.4), as well as the medication on (from 18.6+/-12.1 to 10.1+/-5.9) phase. The UPDRS IVa (dyskinesias) and IVb (motor fluctuations) scores decreased significantly. At long-term follow-up, there were still significant improvements on the total UPDRS III motor score (from 42.3+/-9.3 to 24.2+/-13.2), as well as in all motor subscores, in the off phase, during stimulation. In the on phase, the only significant improvement was seen for rigidity. Complications included hypomania to mania in four patients. Our results indicate that HFS STN results in long-lasting improvement of the motor symptoms, ADL activities and functional performance in patients suffering from advanced PD. The stimulation induced behavioural changes need special consideration. PMID- 15823481 TI - Effect of subthalamic nucleus stimulation patterns on motor performance in Parkinson's disease. AB - We studied different patterns of deep brain stimulation (DBS), but same average rate, in seven Parkinson disease patients performing a wrist flexion/extension task. Movement times were shorter with regular (continuous) stimulation than cycled 'on' and 'off' for 0.1s (mean difference 0.129 s; 95% confidence interval [CI], 0.228-0.029 s; P < 0.007) and tended to be shorter than cycling at 0.5s (mean difference 0.076 s, 95% CI, 0.171 to -0.020 s; P < 0.6). Movement times under stimulation cycling at 0.5 s tended to be shorter than cycling at 0.1 s (mean difference, 0.083 s, 95% CI, 0.188 to -0.022 s, P < 0.12). Any therapeutic mechanisms of action of DBS must account for patterns of stimulation. PMID- 15823482 TI - Early-onset Parkinson's disease in a Chinese population: 99mTc-TRODAT-1 SPECT, Parkin gene analysis and clinical study. AB - Early Onset Parkinson's Disease (EOPD) is characterized by selective degeneration of nigrostriatal dopaminergic neurons and a marked response to levodopa. However, at present, few methods are available as diagnostic tools for EOPD except for 18F DOPA PET. In addition, little is known about the correlation between clinical severity, neuroimaging grading and genetic susceptibility. In the present study, 99mTc-TRODAT-1 SPECT and brain MRI were used to identify 30 cases of non-familial EOPD from a Chinese cohort of 230. All 30 PD patients had an age of onset of less than 55 years (mean age at onset, 41.5+/-9.3 years). Each of the 30 EOPD cases was sub-classified into one of five stages based on the 99mTc-TRODAT-1 SPECT findings. In the early stages of PD (stages 1 and 2), a lower uptake of 99mTc TRODAT-1 in the putamen was found, while uptake in the caudate nucleus was normal. In the latter stages (stages 3, 4, 5), 24 patients revealed a diffuse and uniform loss of 99mTc-TRODAT-1 uptake in the putamen and the caudate nucleus. Further, in conventional genetic studies of the 30 patients, six novel mutations were found in the Parkin gene, and these included five heterozygous point mutations (C441R, Q311H, V258M, C212G, and S193I) and one homozygous deletion (exon 10-12). Known polymorphisms (Ser167Asn, Val380Leu) were also found in a number of patients. However, gene dosage analysis did not reveal any compound heterozygous mutations in these 30 patients using quantitative duplex PCR. This is the first study to examine EOPD patients of Chinese ethnic background (not exhibiting a definite familial trait), to offer a complete genetic analysis of the Parkin gene, and to correlate clinical stages of the disease with dopamine re uptake. PMID- 15823483 TI - Visuo-spatial processing in Parkinson's disease: evidence for diminished visuo spatial sketch pad and central executive resources. AB - A dual-task study compared the visuo-spatial sketch pad and central executive components of working memory as potential cognitive mechanisms of visuo-spatial dysfunction in Parkinson's disease. Mildly to moderately affected Parkinsonian patients (n = 15) and controls (n = 15) performed the Corsi blocks task concurrently with tasks designed to load on the visuo-spatial sketch pad (spatial tapping) or the central executive (random interval repetition). Patients performed more poorly in both concurrent task conditions, implicating a reduction in both visuo-spatial sketch pad and central executive resources. The impact of the concurrent tasks varied with disease severity, with the central executive deficit prominent at disease onset, but the visuo-spatial sketch pad deficit becoming apparent only in the moderate stages of the illness. PMID- 15823484 TI - Paraneoplastic progressive supranuclear palsy syndrome in a patient with B-cell lymphoma. AB - An important component in the diagnosis of atypical parkinsonian disorders is the exclusion of secondary causes. Paraneoplastic causes of parkinsonism are extremely rare. We describe a case which presented initially as probable progressive supranuclear palsy (PSP) but on follow-up displayed a rapidly progressive course, unexplained fever, peripheral neuropathy and an abnormal CSF. We highlight the difficulties faced in formulating a diagnosis for this unusual case prior to the discovery of an occult B-cell lymphoma, and discuss its relevance in the exclusion criteria for PSP. A paraneoplastic cause should be considered if disease progression is unusually rapid. PMID- 15823485 TI - Primary Sjogren's syndrome presenting as a generalized chorea. AB - We report on a patient who presented with generalized chorea as the first manifestation of Sjogren Syndrome, and review the possible pathogenesis. PMID- 15823486 TI - Dystonia after a bone fracture of the arm in a patient with a history of striato pallidal ischemic stroke: a case report. AB - We report on a female with segmental dystonia of the upper limb after an anatomically related ischemic stroke. Dystonia developed almost 3 years after the onset of the stroke and immediately following the removal of the cast because of a bone fracture in the same limb. We discuss the case considering issues such as: delay-onset, lesion topography and pathophysiology, peripheral input and their possible contribution to the development of secondary dystonia. PMID- 15823487 TI - Neurophysiology of blepharospasm and multiple system atrophy: clues to its pathophysiology. PMID- 15823489 TI - Metabolic and cardiovascular disease: complications and consequences. PMID- 15823490 TI - Impact of diabetes/metabolic syndrome in patients with established cardiovascular disease. AB - The significance of the metabolic syndrome (MetS) resides either in its ability to identify individuals at high risk of future disease and disability or in its ability to identify individuals in need of a specific treatment. We have previously shown that in the general population the ability of the MetS to identify individuals at increased risk of diabetes or cardiovascular disease (CVD) is inferior to the ability of established predicting models for these conditions. Although it may someday become routine to recommend treatment with insulin-sensitising agents for non-diabetic individuals with the MetS, most of whom are insulin resistant, there are no clinical trial data to support such a recommendation at the present time. Currently, the treatment of the MetS is based on treatment of its component parts. In the present paper, we examine the role of the MetS as defined by the National Cholesterol Education Program (NCEP) criteria in predicting all-cause and CVD mortality in patients with prevalent CVD from the San Antonio Heart Study (SAHS). This population contains a high proportion of Mexican Americans, who are at high risk of developing type 2 diabetes. After adjusting for age and gender, the MetS is moderately predictive of all-cause and CVD mortality. After further adjustment for diabetes, however, the effect of the MetS becomes non-significant in this population. Moreover, among non-diabetics with prevalent CVD, the MetS was not associated with either all-cause or CVD mortality. Thus, this study indicates that the effect of the MetS on these endpoints is primarily driven by the inclusion in the NCEP definition of diabetes, itself a well-established, potent CVD risk factor. In fact, the prevalence of diabetes in SAHS patients with CVD and the MetS was 42% compared with only 9% in patients with CVD, but without the MetS. This excess prevalence of diabetes appears to account for the enhanced all-cause and CVD mortality in subjects with the MetS. However, these results will need to be confirmed in other populations. PMID- 15823491 TI - Adiponectin: Identification, physiology and clinical relevance in metabolic and vascular disease. AB - Adiponectin is the most abundant adipose-specific protein. Its expression is reduced in obesity, insulin resistance and type 2 diabetes, and plasma concentrations are inversely related to body weight, especially visceral adiposity. Adiponectin is also inversely associated with other traditional cardiovascular risk factors, such as blood pressure, low-density lipoprotein cholesterol and triglyceride levels, and is positively related to high-density lipoprotein cholesterol (HDL-C) levels. Recent research has indicated that adiponectin has anti-inflammatory, anti-atherogenic and antidiabetic properties. The ability of adiponectin to reduce insulin resistance in conjunction with its anti-inflammatory and anti-atherogenic properties makes this novel adipocytokine a promising therapeutic target, and agents that enhance adiponectin secretion or action have potential for treatment of metabolic and vascular disease. Current management strategies that may increase endogenous adiponectin production in humans include weight loss, soy protein and therapy with peroxisome proliferator activating receptor gamma (PPARgamma) agonists. PMID- 15823492 TI - The inflammation: lipoprotein cycle. AB - The central roles played by lipoproteins in atherosclerosis are well established. Increased plasma concentrations of low-density lipoproteins (LDLs) and triglyceride-rich remnant lipoproteins are highly atherogenic, whereas high density lipoproteins (HDLs) are known to protect against lesion development. These effects are driven, in part, by the impact of these lipoproteins on inflammation-a process that is central to atherogenesis. In individuals with dyslipidaemia, LDLs and other atherogenic lipoproteins enter the arterial wall where they undergo chemical modification, including oxidation. These modified lipoproteins initiate the inflammatory process that culminates in atherosclerosis lesion development. The inflammation can be reversed by HDLs via several mechanisms. These include promotion of cholesterol efflux, inhibition of LDL oxidation and reduction of adhesion molecule expression. Recent work has shown that HDLs are also able to inhibit acute vascular inflammation. Given the central roles played by lipoproteins and inflammation in atherogenesis, effective anti atherosclerotic treatments should both modify the lipid profile and target the ongoing inflammation. These criteria are fulfilled by statins, which reduce inflammation by both lipid-dependent and -independent mechanisms. Additional protection from atherosclerosis may be provided by novel therapies that aim to increase plasma levels and activity of HDLs. PMID- 15823493 TI - Markers of inflammation and their clinical significance. AB - Inflammation plays an important role in the initiation and progression of atherosclerosis and the development of atherosclerotic events. Understanding the molecular basis of inflammation has led to the identification of markers that may also serve as new targets of therapy in the management of atherothrombotic disease. Inflammatory markers, such as C-reactive protein (CRP), have been shown to predict future cardiovascular events in individuals with and without established cardiovascular disease (CVD). Statins substantially reduce cardiovascular morbidity and mortality, and recently their anti-inflammatory properties have been investigated. In this paper, we discuss biomarkers implicated in the inflammatory process leading to atherothrombosis, including CRP, adiponectin, monocyte chemoattractant protein 1 (MCP-1), CD40 ligand and lipoprotein-associated phospholipase A(2) (Lp-PLA(2)), and the effect of statins on these markers and their potential relationship to cardiovascular events. PMID- 15823494 TI - Target gene discovery in extended families with type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a polygenic, multifactorial disease. Multiple attempts have been made to identify genes that predispose to development of diabetes using both the candidate gene approach and whole genome scanning. However, in spite of substantial financial commitment, very few targets have been identified. Identification of predisposing genes is complicated by several factors. Diabetic patients demonstrate a variety of gene defect combinations. Identification of predisposing genomic regions is thus impeded if data are collected from a heterogeneous population. In addition, the diseased phenotype may only manifest when the net effect of the predisposing factors exceeds a certain "threshold". The effects of predisposing genetic and environmental factors thus appear to be additive, and this also complicates target gene discovery. Using the traditional approach, significant associations between genomic regions and disease are rarely observed unless data are acquired from hundreds of individuals. In contrast, results from whole genome scans performed in homogeneous and consanguineous populations with a high incidence of type 2 diabetes have demonstrated that highly significant associations can be obtained using data from a small number of subjects belonging to the same extended family. Such populations offer the promise of substantial progress in type 2 diabetes genomics research. PMID- 15823495 TI - Diabetic dyslipidaemia: effective management reduces cardiovascular risk. AB - Patients with diabetes are at significantly increased risk for coronary heart disease (CHD); even those patients without a history of a previous myocardial infarction (MI) have as high a risk of a fatal or nonfatal MI as nondiabetic patients with a history of previous MI. As a result it is now generally recommended that cardiovascular risk factors be treated as aggressively in patients with diabetes as in nondiabetic patients with a history of CHD. Results from the recently published Collaborative Atorvastatin Diabetes Study (CARDS) and meta-analysis of primary and secondary interventions trials confirm that there is a uniform relative risk reduction across a wide range of high-risk patients including diabetes patients without established CHD. A highly significant 22-24% reduction in risk of future vascular events is evident when patients with diabetes are treated with statins in trials. Current guidelines, including the recently updated National Cholesterol Education Program (NCEP) guidelines, endorse aggressive, early intervention in very-high-risk patients, such as those with diabetes plus cardiovascular disease (CVD), regardless of baseline low density lipoprotein cholesterol (LDL-C) level in order to achieve an LDL-C goal of 70 mg/dL (1.8 mmol/L). However, despite increasing evidence and knowledge of the value of lipid lowering, a recent survey of diabetes specialists indicates that many patients with diabetes remain untreated or undertreated. The availability of more effective statins should help to close this "action gap", in concert with other measures such as initiatives to improve patient compliance. PMID- 15823496 TI - Proteasomal processing of nuclear autoantigens in systemic autoimmunity. AB - Autoimmune diseases are believed to rise from a breakdown of tolerance and a deregulation of the immune system to discriminate effectively between "self" and "non-self". Although the origins of autoimmunity and the factors leading to overt autoimmune diseases are largely unclear, genetic background, hormones, and environmental agents are found to be involved. The production of autoantibodies against distinct "self"-proteins of the cell nucleus constitutes one of the autoimmune manifestations that characterize systemic autoimmunity. While molecular mechanisms of anti-nuclear autoimmune responses remain poorly understood, processing and presentation of intracellular autoantigens to T cells constitutes a central issue in the generation of antigen-driven autoimmunity. Intracellular proteins, including nuclear autoantigens, are degraded by the proteasomal pathway that clears away endogenous proteins, regulates numerous cellular processes, and delivers immunocompetent peptides to the antigen presenting machinery. Since antigen processing is a key step for the maintenance of peripheral tolerance, in this article, we discuss the role of proteasome dependent antigen processing in systemic autoimmunity. PMID- 15823498 TI - Epidemiology of adult rheumatoid arthritis. AB - Several incidence and prevalence studies of rheumatoid arthritis (RA) have been reported during the last decades, suggesting a considerable variation of the disease occurrence among different populations. The majority of studies carried out in Northern European and North American areas estimate a prevalence of 0.5 1%, and a mean annual incidence of 0.02-0.05%. The occurrence of the disease seems to be lower in other parts of the world. Some studies from North American, North European, and Japanese populations suggest a decline in both the prevalence and incidence of the disease after the 1960s. RA is related to an increased mortality, and the expected survival of RA patients is likely to decrease 3-10 years. There is epidemiological evidence that genetic factors are related to an increased risk of RA. However, RA is considered to be a multifactorial disease, resulting from the interaction of both genetic and environmental factors, which contribute to its occurrence and expression. The main risk factors for the disease include genetic susceptibility, sex and age, smoking, infectious agents, hormonal, dietary, socioeconomic, and ethnic factors. Most of these factors are likely to be associated with both disease occurrence and severity. PMID- 15823497 TI - Statins and autoimmune diseases. AB - Inhibitors of 3-hydroxy-3methylglytaryl coenzyme A (HMG-CoA) reductase or statins are effective lipid lowering drugs widely used in cardiovascular disease. In the recent years, pleotropic effects of statins have been reported, which include anti-inflammatory and immunomodulatory properties. This review discusses the anti inflammatory and immunomodulatory roles of statins and their possible use for the treatment of other inflammatory diseases or conditions with the involvement of the immune system. PMID- 15823499 TI - The immunobiology of primary sclerosing cholangitis. AB - An understanding of the immunobiology of primary sclerosing cholangitis (PSC) is essential to improving both diagnosis and treatment. There have been significant gains in the discovery of genetic polymorphisms that generate susceptibility to disease, but only limited data on etiologic events that may initiate the inflammatory response. Colonic inflammation produces memory T cells that have the ability to bind both biliary and colonic endothelial cells. One possible mechanism for the development of PSC is the homing of these memory T cells to the biliary tree. In addition, TNF(alpha) may contribute to the oxidative damage of the biliary system. Finally, although speculative, mononuclear cell responses against biliary epithelial cells may create a persistent inflammatory response, eventually leading to fibrosis. PMID- 15823500 TI - Potential off-label use of infliximab in autoimmune and non-autoimmune diseases: a review. AB - TNF-alpha is a crucial cytokine in the establishment and maintenance of inflammation in multiple autoimmune and non-autoimmune disorders. A number of large placebo-controlled trials have shown that infliximab, a chimeric monoclonal antibody against TNF-alpha, is effective and well-tolerated in patients with Crohn's disease and rheumatoid arthritis (RA) and has become a widely used treatment for these diseases. More recent controlled trials have also shown the effectiveness of TNF-alpha blockers in psoriasis, psoriatic arthritis, and ankylosing spondylitis. The results of clinical trials, open-label studies, and case studies indicate that TNF inhibitors (alone or in combination with other protocols) look very promising for the treatment of a variety of other conditions, including uveitis, sarcoidosis, Sjogren's syndrome (SS), Behcet's syndrome, vasculitis, and graft versus host disease. There is a rationale for using TNF blockade even in systemic lupus erythematosus, a prototype of autoantibody-mediated disease, and a pilot study seems to confirm this potential effective approach. The neutralisation of TNF might therefore play a role in the treatment of many autoimmune and non-autoimmune disorders other than Crohn's disease or RA. We here review the current and prospective roles of infliximab in the treatment of autoimmune diseases and other conditions that do not currently have FDA or EMEA approval. PMID- 15823501 TI - TNF-alpha, rheumatoid arthritis, and heart failure: a rheumatological dilemma. AB - Cardiovascular disease (CVD) is responsible for 35-50% of rheumatoid arthritis (RA) deaths, whereas, in the general UK adult population, coronary heart disease is responsible for 1/4 deaths in males and 1/5 deaths in female. This increased risk may be attributable to RA-specific risk factors such as hyperhomocysteinemia, disease-related dyslipidemia or vascular inflammation, or to morbidity related to medications and high levels of tumor necrosis factor alpha (TNF-alpha). The possible roles of TNF-alpha in the development of atherosclerosis include the recruitment of inflammatory cells to the site of injury or the promotion of adverse vascular smooth muscle cell remodelling. TNF alpha may also act as a proinflammatory factor in plaque rupture. Anticytokine therapy could prove beneficial in the treatment of patients with heart failure. While early studies supported this hypothesis, anti-TNF strategies have not demonstrated salutary benefits in large multicenter randomized and placebo controlled clinical trials in patients with symptomatic heart failure. There is a variety of possible explanations for the failure of anti-TNF therapy: (1) TNF antagonism has untoward effects in the setting of heart failure; (2) the biological agents used in the trials were intrinsically toxic; (3) sex and race may have important implications in the outcome after anticytokine therapy; (4) the TNF-alpha protein contains a polymorphism, and, in fact, genoma plays a role in modifying the pharmacologic response to anticytokines; (5) anti-TNF-alpha approaches could have had pharmacodynamic interactions with other heart failure medications; and (6) the patients in these trials may have been inappropriately selected. These disappointing results may determine controversial attitude in the long-term treatment with anti-TNF agents in RA or Crohn's disease. The effects of TNF-alpha blockers on incident cases of congestive heart failure (CHF) in RA are controversial. The available published data suggest the following: (a) RA patients with history of CHF and a concomitant indication for the use of TNF alpha blockers do not need a baseline cardiac evaluation to screen for heart failure; (b) patients with well-compensated mild CHF New York Heart Association (NYHA) classes I and II and a concomitant indication for the use of TNF-alpha blockers should be evaluated at baseline and then be closely monitored for any clinical signs of worsening heart failure; and (c) patients with (NYHA) class III or IV heart failure should not be treated with TNF-alpha blockers in any case. PMID- 15823502 TI - Safety of tumour necrosis factor and interleukin-1 blocking agents in rheumatic diseases. AB - The three licensed TNF(alpha) blocking agents (etanercept, infliximab, adalimumab) and the recombinant form of human interleukin-1-receptor antagonist (anakinra) have all been shown to be effective in patients with chronic rheumatic autoimmune diseases; they have also been associated with certain types of serious adverse events. As expected, much of the information on serious events have accumulated during the post-marketing period. Certain serious, but uncommon, adverse events have been observed with all three TNF(alpha) blocking agents, including serious bacterial infections, tuberculosis (TB) and certain opportunistic infections, demyelinating syndromes, and lupus-like reactions. These data suggest that these adverse reactions may be related to blockade of TNF(alpha) and may therefore represent class effects of these agents. However, the severity and degree of risk may not be the same with all three agents. Blockade of interleukin-1 activity with anakinra appears, at present, to be relatively safe. The safety profile of these products will continue to be developed through the use of the registry, periodic safety updates from the passive surveillance program, and safety data from controlled trials of biological therapy for other diseases. Physicians should minimize risks by patient selection and screening for opportunistic infections. Moreover, the choice of the biological agent must be tailored to minimize risks and maximize benefits. PMID- 15823503 TI - The role of echocardiographic techniques in connective tissue diseases. AB - Connective tissue diseases (CTD) lead to a high prevalence of common cardiac manifestations (pericarditis and myocarditis) and of ischemic coronary events with a considerable increase in cardiac mortality related to premature atherosclerosis. Although there are several techniques able to detect cardiac involvement in CTD patients, the most useful and non-invasive technique is echocardiographic exam which is able to detect not only valvular abnormalities, pericardial diseases and pulmonary hypertension but also left ventricular (LV) systolic or diastolic (regional or global) wall motion dysfunction. It is also well known that transesophageal echocardiography (TEE) can better identify cardiac abnormalities, vegetations and embolic sources. Symptomatic patients with positive stress echocardiographic exam or dipyridamole thallium imaging test should be referred for possible cardiac catheterization, especially if a large ischemic territory is involved. Until now, the echocardiographic evaluation of coronary artery tree consisted of assessing regional and global left ventricular systolic and diastolic function at rest and during pharmacological stress test. Recently, a new echocardiographic noninvasive method that allows direct assessment of coronary flow velocity in the mid-distal portion of left anterior descending artery (LAD) has been developed and validated. Advanced ultrasound technology (high-frequency broadband transducer with second harmonic capability) has now made possible a direct arterial visualization and measurement of coronary artery flow in left anterior descending in CTD patients with the assessment of coronary flow reserve (CFR). PMID- 15823504 TI - From animal models to human genetics: research on the induction and pathogenicity of autoantibodies. AB - The revolutionary techniques of modern molecular and cellular biology enhance almost daily our knowledge of immunity and autoimmunity in men and experimental animals. Our fragmentary puzzle of the immune system is going to form a fascinating picture of a masterpiece of evolution. Although many of these aspects were achieved by analysis of human body fluids and tissues, the etiopathogenesis of autoimmune diseases cannot readily be analyzed without appropriate animal models. Therefore, the 7th Dresden Symposium on Autoantibodies has focused on experimental autoimmune models. The 295 attendants of the symposium listened to and discussed about the pathogenesis and therapy of autoimmunity in experimental mouse models, natural and pathogenic autoantibodies, molecular mechanisms of xenobiotic-induced autoimmunity, the genetic background of autoimmune diseases, novel autoantibodies and their pathogenic and/or clinical relevance, autoantibodies in systemic and neurological diseases, the occurrence and measurement of therapy-induced antibodies and methodical aspects as well as novel diagnostic strategies including multiplex assays for autoantibody profiling. Those who are interested to read the full length articles are referred to the book published in parallel to this meeting ([Conrad K, Bachmann MP, Chan EKL, Fritzler MJ, Humbel RL, Sack U, Shoenfeld Y, editors. From animal models to human genetics: research on the induction and pathogenicity of autoantibodies, Report on the 7th Dresden Symposium on Autoantibodies held in Dresden on September 1-4, 2004. Germany: Pabst Science Publishers; 2004.]; www.pabst-publishers.de). PMID- 15823505 TI - Differentially expressed nucleolar TGF-beta1 target (DENTT) shows tissue-specific nuclear and cytoplasmic localization and increases TGF-beta1-responsive transcription in primates. AB - Differentially Expressed Nucleolar TGF-beta1 Target (DENTT) is a new member of the TSPY/TSPY-like/SET/NAP-1 (TTSN) superfamily whose mRNA is induced by TGF beta1 in TGF-beta1-responsive human lung cancer cells. Monkey DENTT mRNA contains a 2085-bp open reading frame that encodes a predicted polypeptide of 695 amino acids with five nuclear localization signals, two coiled-coil regions, and a domain that shows significant identity to a region that defines the TTSN superfamily. RT-PCR amplification and Western blot analyses showed DENTT mRNA and protein in adult monkey tissues, including the adrenal gland, cerebral cortex, and ovary. Immunohistochemical staining showed that numerous neurons were intensely immunoreactive for DENTT, as were anterior pituitary secretory cells, thyroid follicular cells, and smooth muscle cells of arteries and lung bronchial walls. DENTT expression was also prominent in monkey bronchiolar-alveolar adenomas and cell lines. While the addition of TGF-beta1 or retinoic acid to monkey normal lung bronchial 12MBr6 cells and human lung cancer NCI-H727 cells increased DENTT protein production, TGF-beta1 together with retinoic acid resulted in a more sustained increase in DENTT production than with TGF-beta1 or retinoic acid alone. Transient transfection studies showed that ectopic DENTT expression significantly increased TGF-beta1-responsive 3TP-Lux and CAGA12-Lux reporter transcription in 12MBr6 and NCI-H727 cells with TGF-beta1 addition, while ectopic DENTT expression had no significant effect on the transcription of a retinoic acid-responsive element reporter in the presence of retinoic acid or TGF-beta1. These findings suggest new possibilities for DENTT as a TGF-beta1 regulated, but not a retinoic acid-regulated member of the TTSN superfamily in primate physiology. PMID- 15823506 TI - Biochemical observation of the rapid mobility of nuclear HMGB1. AB - Formaldehyde-crosslinked and sonicated chromatin fragments were obtained from 15 day chicken embryo erythrocytes and purified on caesium chloride gradients. Polyclonal antibodies raised against chicken HMGB1 were used to immuno precipitate fragments carrying HMGB1 in two protocols: (1) affinity purified antibodies covalently coupled to agarose beads and (2) diluted antiserum. The DNA of the antibody-bound chromatin was quantified and its sequence content assessed by quantitative real-time PCR to give values of the absolute enrichments generated. Amplicons were monitored within the active beta-globin locus, in the adjacent heterochromatin, in the lysozyme locus (containing an active housekeeping gene and the inactive lysozyme gene) and at the promoter of the inactive ovalbumin gene. For all amplicons the Bound/Input ratio was close to unity, implying no preferential location of HMGB1 on the chromatin. This initially unexpected result can now be understood in the light of the exceptional mobility of HMGB1 revealed by FLIP experiments showing that only 1-2 s are needed for HMGB1 to cross the nucleus: crosslinking times of 1 min were used in the present experiments. PMID- 15823507 TI - Theaflavin derivatives in black tea and catechin derivatives in green tea inhibit HIV-1 entry by targeting gp41. AB - Theaflavin derivatives and catechin derivatives are the major polyphenols in black tea and green tea, respectively. Several tea polyphenols, especially those with galloyl moiety, can inhibit HIV-1 replication with multiple mechanisms of action. Here we showed that the theaflavin derivatives had more potent anti-HIV-1 activity than catechin derivatives. These tea polyphenols could inhibit HIV-1 entry into target cells by blocking HIV-1 envelope glycoprotein-mediated membrane fusion. The fusion inhibitory activity of the tea polyphenols was correlated with their ability to block the formation of the gp41 six-helix bundle, a fusion active core conformation. Computer-aided molecular docking analyses indicate that these tea polyphenols, theaflavin-3,3'-digallate (TF3) as an example, may bind to the highly conserved hydrophobic pocket on the surface of the central trimeric coiled coil formed by the N-terminal heptad repeats of gp41. These results indicate that tea, especially black tea, may be used as a source of anti-HIV agents and theaflavin derivatives may be applied as lead compounds for developing HIV-1 entry inhibitors targeting gp41. PMID- 15823508 TI - The cellulose synthase gene PrCESA10 is involved in cellulose biosynthesis in developing tracheids of the gymnosperm Pinus radiata. AB - One full length (PrCESA10) and seven other cDNA clones (PrCESA2, 3, 5-8, 11) encoding cellulose synthases (CESAs) were isolated from the coniferous gymnosperm Pinus radiata. PrCESA10 encodes a protein predicted to contain the same domains and regions as angiosperm CESA genes: a zinc finger domain, a hypervariable region 1 (HVR1), a plant-conserved region (CR-P), a class-specific region or hypervariable region 2 (HVR2), in addition to the four conserved domains U1-U4 that are characteristic of the family 2 processive beta-glycosyltransferases. The P. radiata protein is also predicted to contain eight transmembrane domains. The zinc finger domain, the CR-P and the C-terminal portion of the proteins, are highly conserved between P. radiata and the nearest angiosperm CESA protein from Solanum tuberosum. Reverse transcriptase-PCR showed that all the P. radiata genes were expressed in all organs tested, although to different extents. In situ hybridization studies with PrCESA10 in stems of 2- and 12-month-old seedlings showed that it was expressed in the secondary xylem in the two-to-three most recently developed tracheids, which were laying down secondary cell walls. PMID- 15823509 TI - The expression of de novo DNA methylase DNMT3b, of the methyl-CpG binding protein MBD2b and of 5-MCDG glycosylase shows two waves of induction during CaCO-2 cell differentiation. AB - DNA methylation plays a central role in the control of gene expression during development and cell differentiation, thus DNA methylation and demethylation processes are expected to be strictly regulated during these events. We have explored the expression levels of the genes encoding DNA methylases, methyl-CpG binding proteins and demethylases during in vitro differentiation of human carcinoma colon cells (CaCO-2) used as a model system. The results show that the global DNA methylation pattern remains constant during CaCO-2 cells differentiation indicating that required genome methylation pattern in cell differentiation was already established in the seeded cells. On the contrary, the timing of expression of several of the explored genes is tightly regulated, suggesting they are involved in the regulation of the differentiation program. In particular, the timing of expression of DNMT3b and of MBD2b and 5-MCDG shows two peaks not observed in the time courses of the expression of other genes belonging to the same families. These events, not dependent on the cell cycle synchronization, have apparently no significant impact on the overall methylation status of the genome. PMID- 15823510 TI - Effect of levetiracetam on the pharmacokinetics of adjunctive antiepileptic drugs: a pooled analysis of data from randomized clinical trials. AB - The purpose of this study was to determine the influence of levetiracetam on the steady-state serum concentrations of other commonly used antiepileptic drugs (AEDs). Serum AED concentrations were measured at baseline and after adjunctive therapy with levetiracetam (1000-4000 mg/day) or placebo in four phase III trials in patients with refractory partial epilepsy receiving stable AED dosages. The data were pooled, and repeated measures covariance analysis was used to calculate the ratio (and 90% confidence intervals) of the geometric mean serum drug concentrations during adjunctive levetiracetam therapy relative to baseline. Levetiracetam did not increase or decrease mean steady-state serum concentrations of carbamazepine, phenytoin, valproic acid, lamotrigine, gabapentin, phenobarbital, or primidone. For each of these AEDs, the 90% confidence interval of the geometric mean drug concentrations ratio was included within the 80-125% bioequivalence range. Serum concentrations of these AEDs did not change over time after adjunctive levetiracetam therapy, irrespective of the dosage of levetiracetam used. For vigabatrin, there was no evidence for a significant change in serum drug concentration after the addition of levetiracetam, but the number of observations was too small for the limits of the confidence interval to fall within the 80-125% range. Thus, adjunctive therapy with levetiracetam does not influence the steady-state serum concentrations of concomitantly administered carbamazepine, phenytoin, valproic acid, lamotrigine, gabapentin, phenobarbital, or primidone. Consequently, no need for adjusting the dosages of these AEDs is anticipated when levetiracetam is added on or removed from a patient's therapeutic regimen. PMID- 15823511 TI - Sensing cell stress and transformation through Vgamma9Vdelta2 T cell-mediated recognition of the isoprenoid pathway metabolites. AB - Vgamma9Vdelta2 cells, a major peripheral blood gammadelta T cell subset in adults, recognize non-peptidic phosphorylated metabolites referred to as phosphoantigens (phosphoAg), which are produced by a broad array of prokaryotic and eukaryotic organisms. We will review here the biosynthetic pathways leading to production of phosphoAg and our current understanding of the mode of activation of Vgamma9Vdelta2 cells by these compounds. We will also discuss the physiological relevance of this immune recognition process and show how it can enable discrimination by Vgamma9Vdelta2 lymphocytes of infected and/or transformed cells. PMID- 15823512 TI - The establishment of Neisseria meningitidis serogroup W135 of the clonal complex ET-37/ST-11 as an epidemic clone and the persistence of serogroup A isolates in Burkina Faso. AB - We analyzed 48 invasive isolates of Neisseria meningitidis that were isolated from meningitis cases in Burkina Faso (April 2002 to April 2003). Thirty-nine of these isolates had the phenotype (serogroup:serotype:serosubtype) W135:2a:P1.5,2, eight isolates were A:4:P1.9 and one isolate was nongroupable:nonserotypable:nonserosubtypable. Genotyping of meningococcal isolates showed that W135 isolates belonged to the sequence type (ST)-11. The nongroupable isolate was of genogroup W135 and belonged to ST-192. Isolates of serogroup A belonged to ST-2859 (a member of the subgroup III/ST-5 clonal complex). W135 (ST-11) isolates involved in meningitis outbreaks in Burkina Faso differed from those involved in the Hajj-2000 associated outbreak by their pulsed field gel electrophoresis profile. These data confirm the changing epidemiology of meningococcal infection in Burkina Faso with the establishment and expansion of serogroup W135 N. meningitidis strains of the ET-37/ST-11 clonal complex, as well as the emergence of a new clone within the subgroup III/ST-5 clonal complex. PMID- 15823514 TI - Role of CD8+ T lymphocytes in control of Mycobacterium tuberculosis infection. AB - Tuberculosis remains a global health concern. Control of infection is dependent on cell-mediated immune responses, with CD4+ T lymphocytes playing a central role. In this article, data supporting the importance of CD8+ T lymphocytes is reviewed, with an emphasis on the unique functional roles that this lymphocyte subset may play. PMID- 15823513 TI - Properties of anti-gp41 core structure antibodies, which compete with sera of HIV 1-infected patients. AB - To determine the correlation between the immunoreaction against the core structure of human immunodeficiency virus type (HIV-1) transmembrane protein gp41 epitopes and the disease progression, it is essential to evaluate the anti-core structure antibody epitopes and the humoral immunity against the epitopes. For this purpose we evaluated monoclonal antibodies (mAbs) against the gp41 core structure such as mAbs 50.69, 98.6 and T26, by Western blotting (WB) and flow cytometry. WB showed mAbs 50.69 and 98.6 bound to both monomeric and oligomeric gp41, and mAb T26 exclusively bound to oligomeric gp41. We evaluated the sera from Pneumocystis pneumonia patients (PCP; n=7) and long-term survivors (LTS; n=7). Competition assay with sera and mAbs for binding to H9 cells infected with HIV-1 IIIB virus was done using flow cytometry. The results revealed that PCP sera as well as LTS sera inhibited the binding of all the three mAbs, and the PCP sera inhibited mAb T26 binding more efficiently than LTS. Therefore, PCP patients retain competing immunity to antibodies against not only the shared epitopes of the core structure (binding sites of mAbs 50.69 and 98.6) but also against oligomeric gp41 specific epitope (binding site of mAb T26). PMID- 15823515 TI - The multitude of targets for the immune system and drug therapy in the fungal cell wall. AB - Recent studies on fungi revealed that several cytosolic and membrane components migrate to the cell wall together with secreted proteins and biosynthetic polysaccharides to build a dynamic immunoreactive structure. New aspects of fungal cell wall assembly and biosynthesis, focusing on the potential of glycolipids, melanin, heat-shock proteins, histone and surface antigens as targets of drugs and antifungal antibodies are discussed. PMID- 15823516 TI - Differential contribution of sodC1 and sodC2 to intracellular survival and pathogenicity of Salmonella enterica serovar Choleraesuis. AB - Several of the most virulent Salmonella enterica strains possess two genes encoding periplasmic Cu,Zn superoxide dismutase, sodC1 and sodC2, located on a lambdoid prophage and on the chromosome, respectively. These genes contribute to Salmonella virulence by protecting bacteria from superoxide generated by the host's phagocytes. To investigate the respective contributions of sodC1 and sodC2 to the virulence of a clinical isolate of Salmonella enterica serovar Choleraesuis (S. choleraesuis), we have analyzed both the intracellular survival of wild type and sodC mutant strains within J774 macrophages and Caco-2 cells, and their ability to proliferate in intraperitoneally-infected mice in competition assays. In agreement with previous studies, mutant strains lacking one or both sodC genes were equally impaired in their ability to survive within activated macrophages. However, when macrophage killing experiments were carried out with non-opsonized bacteria, sodC2 contributed to intracellular survival more than sodC1, indicating that changes in the pathways of bacterial uptake can modify the relative role of the two sodC genes. More unexpectedly, we have found that the ability of S. choleraesuis to survive within Caco-2 cells was severely affected by inactivation of sodC genes, sodC2 being more important than sodC1. As Caco-2 cells actively produce superoxide, this suggests that oxygen radical production by colonic cells has a role in controlling proliferation of facultative intracellular bacteria. Mouse infection studies confirmed that, in the S. choleraesuis strain under investigation, both sodC genes are required to confer full virulence, sodC2 contributing slightly more than sodC1 to Salmonella pathogenesis. Our findings contrast with the results of other studies carried out in S. enterica serovar Typhimurium and suggest that the relative contributions of sodC1 and sodC2 to host-pathogen interactive biology may vary depending on the Salmonella serovar or strain. PMID- 15823518 TI - Intestinal microbiota. PMID- 15823517 TI - Maturity- and sex-related changes in tibial bone geometry, strength and bone muscle strength indices during growth: a 20-month pQCT study. AB - During growth, bone strength is conferred through subtle adaptations in bone mass and geometry in response to muscle forces. Few studies have examined the changes in bone geometry, strength and the bone-muscle strength relationship across maturity in boys and girls. Our aims were to describe (i) 20-month changes in bone geometry and strength at the tibial midshaft across three maturity groups of boys and girls, (ii) differences in these adaptations between sexes at the same approximate level of maturity and (iii) the bone-muscle strength relationship across maturity groups of boys and girls and between sexes. We used peripheral quantitative computed tomography (pQCT, Stratec XCT-2000) to measure change in total bone cross-sectional area (ToA, mm(2)), cortical area (CoA, mm(2)), average cortical thickness (C.Th., mm), section modulus (mm(3)) and muscle cross sectional area (mm(2)) at the tibial midshaft (50% site) in 128 EARLY-, PERI- and POST-pubertal girls (n = 69, 11.9 +/- 0.6 years) and boys (n = 59, 12.0 +/- 0.6 years) across 20 months. We also calculated two bone-muscle strength indices (BMSI) for compression (CoA/MCSA) and bending [strength index/MCSA; where strength index = Z / (tibial length / 2)]. EARLY boys and girls had smaller ToA at baseline than same sex PERI or POST participants. There were no sex differences in ToA or CoA at baseline; however, boys increased both parameters significantly more than girls in every maturity group (8.5-11.1%, P < 0.01). These changes in bone geometry conferred greater gains in bone strength for boys compared with girls in each maturity group (13.8-15.6%, P < 0.01). Baseline BMSIs did not differ between sexes for EARLY and PERI groups, whereas BMSIs were significantly higher for POST boys compared with POST girls (P < 0.05). BMSIs decreased for EARLY and PERI girls (-7.4-(-1.1%)) whereas the ratios remained stable for EARLY and PERI boys (-0.6-2.5%). This sex difference in BMSI change was due to a relatively greater increase in CoA among EARLY and PERI boys compared with same-maturity girls. BMSIs remained stable in POST girls and decreased in POST boys due to relatively greater gains in MCSA. This study provides novel longitudinal descriptions of the maturity- and sex-specific changes in bone geometry, strength and bone-muscle strength indices. PMID- 15823519 TI - The real 'domains' of life. PMID- 15823520 TI - Laterality in a non-visual sensory modality--the lateral line of fish. PMID- 15823521 TI - Head-bobbing behavior in foraging whooping cranes favors visual fixation. PMID- 15823522 TI - Insect serosa: a head line in comparative developmental genetics. AB - A recent study reveals specific functions of Hox3/zerknullt (zen) in the extraembryonic and embryonic primordia of the red flour beetle, Tribolium castaneum. The results shed light on the functional evolution of bicoid, a zen paralogue of higher flies, which determines anterior body parts. PMID- 15823523 TI - Neuroeconomics: the shadow of the future. AB - Humans and other animals tend to disregard future benefits and costs when choosing between immediate and delayed gratification. This tendency can lead to the choice of options that are not in one's own long-term interest. A new study looks at the neurophysiological basis of this self-defeating behavior. PMID- 15823524 TI - Sex chromosome inactivation: the importance of pairing. AB - In mammals, the process of making sperm is marked by inactivation of sex chromosomes. Why and how does this happen? The answer apparently lies in whether a chromosome finds a pairing partner. Similar mechanisms in mold and worms reveal a surprising and recurrent theme throughout evolution. PMID- 15823525 TI - Cell fusion: EFF is enough. AB - Developmentally programmed cell-cell fusion in Caenorhabditis elegans requires the EFF-1 protein, which is sufficient to cause normally non-fusing cells to fuse. EFF-1 localizes to fusion-fated membranes, implicating it as a direct fusogen. PMID- 15823526 TI - Social signals: the secret language of mice. AB - Animals are known to produce substances that modulate social and sexual behavior of conspecifics, but the mechanistic details underlying these phenomena have been elusive. A recent paper identifies a male-specific compound in mouse urine that activates olfactory bulb neurons and mediates behavioral attraction. PMID- 15823527 TI - SUMO modification: wrestling with protein conformation. AB - SUMO modification of human thymine-DNA glycosylase facilitates the processing of base excision repair substrates by an unusual mechanism: while leaving the catalytic center unaffected, it induces product release by eliciting a conformational change in the enzyme. PMID- 15823528 TI - Bacterial mitosis: actin in a new role at the origin. AB - MreB is a prokaryotic homolog of actin involved in cellular organization and chromosome segregation. Recent results suggest that MreB is part of a kinetochore like complex that specifically segregates the replication origin region of the bacterial chromosome. PMID- 15823529 TI - Neurotrophins: a ticket to ride for BDNF. AB - The secretion of BDNF from neurons is under activity-dependent control: this is crucial for the formation of appropriate synaptic connections during development and for learning and memory in adults. New evidence shows that interaction between a motif in the tertiary structure of BDNF and the sorting receptor carboxypeptidase E directs this neurotrophin to the regulated secretory pathway. PMID- 15823530 TI - Condensins: organizing and segregating the genome. AB - Condensins are multi-subunit protein complexes that play a central role in mitotic chromosome assembly and segregation. The complexes contain 'structural maintenance of chromosomes' (SMC) ATPase subunits, and induce DNA supercoiling and looping in an ATP-hydrolysis-dependent manner in vitro. Vertebrate cells have two different condensin complexes, condensins I and II, each containing a unique set of regulatory subunits. Condensin II participates in an early stage of chromosome condensation within the prophase nucleus. Condensin I gains access to chromosomes only after the nuclear envelope breaks down, and collaborates with condensin II to assemble metaphase chromosomes with fully resolved sister chromatids. The complexes also play critical roles in meiotic chromosome segregation and in interphase processes such as gene repression and checkpoint responses. In bacterial cells, ancestral forms of condensins control chromosome dynamics. Dissecting the diverse functions of condensins is likely to be central to our understanding of genome organization, stability and evolution. PMID- 15823531 TI - Single units in the pigeon brain integrate reward amount and time-to-reward in an impulsive choice task. AB - BACKGROUND: Animals prefer small over large rewards when the delays preceding large rewards exceed an individual tolerance limit. Such impulsive choice behavior occurs even in situations in which alternative strategies would yield more optimal outcomes. Behavioral research has shown that an animal's choice is guided by the alternative rewards' subjective values, which are a function of reward amount and time-to-reward. Despite increasing knowledge about the pharmacology and anatomy underlying impulsivity, it is still unknown how the brain combines reward amount and time-to-reward information to represent subjective reward value. RESULTS: We trained pigeons to choose between small, immediate rewards and large rewards delivered after gradually increasing delays. Single-cell recordings in the avian Nidopallium caudolaterale, the presumed functional analog of the mammalian prefrontal cortex, revealed that neural delay activation decreased with increasing delay length but also covaried with the expected reward amount. This integrated neural response was modulated by reward amount and delay, as predicted by a hyperbolical equation, of subjective reward value derived from behavioral studies. Furthermore, the neural activation pattern reflected the current reward preference and the time point of the shift from large to small rewards. CONCLUSIONS: The reported activity was modulated by the temporal devaluation of the anticipated reward in addition to reward amount. Our findings contribute to the understanding of neuropathologies such as drug addiction, pathological gambling, frontal lobe syndrome, and attention-deficit disorders, which are characterized by inappropriate temporal discounting and increased impulsiveness. PMID- 15823533 TI - Functionality of human thymine DNA glycosylase requires SUMO-regulated changes in protein conformation. AB - BACKGROUND: Base excision repair initiated by human thymine-DNA glycosylase (TDG) results in the generation of abasic sites (AP sites) in DNA. TDG remains bound to this unstable repair intermediate, indicating that its transmission to the downstream-acting AP endonuclease is a coordinated process. Previously, we established that posttranslational modification of TDG with Small Ubiquitin-like MOdifiers (SUMOs) facilitates the dissociation of the DNA glycosylase from the product AP site, but the underlying molecular mechanism remained unclear. RESULTS: We now show that upon DNA interaction, TDG undergoes a dramatic conformational change, which involves its flexible N-terminal domain and accounts for the nonspecific DNA binding ability of the enzyme. This function is required for efficient processing of the G.T mismatch but then cooperates with the specific DNA contacts established in the active site pocket of TDG to prevent its dissociation from the product AP site after base release. SUMO1 conjugation to the C-terminal K330 of TDG modulates the DNA binding function of the N terminus to induce dissociation of the glycosylase from the AP site while it leaves the catalytic properties of base release in the active site pocket of the enzyme unaffected. CONCLUSION: Our data provide insight into the molecular mechanism of SUMO modification mediated modulation of enzymatic properties of TDG. A conformational change, involving the N-terminal domain of TDG, provides unspecific DNA interactions that facilitate processing of a wider spectrum of substrates at the expense of enzymatic turnover. SUMOylation then reverses this structural change in the product bound TDG. PMID- 15823532 TI - NMDA receptors mediate olfactory learning and memory in Drosophila. AB - BACKGROUND: Molecular and electrophysiological properties of NMDARs suggest that they may be the Hebbian "coincidence detectors" hypothesized to underlie associative learning. Because of the nonspecificity of drugs that modulate NMDAR function or the relatively chronic genetic manipulations of various NMDAR subunits from mammalian studies, conclusive evidence for such an acute role for NMDARs in adult behavioral plasticity, however, is lacking. Moreover, a role for NMDARs in memory consolidation remains controversial. RESULTS: The Drosophila genome encodes two NMDAR homologs, dNR1 and dNR2. When coexpressed in Xenopus oocytes or Drosophila S2 cells, dNR1 and dNR2 form functional NMDARs with several of the distinguishing molecular properties observed for vertebrate NMDARs, including voltage/Mg(2+)-dependent activation by glutamate. Both proteins are weakly expressed throughout the entire brain but show preferential expression in several neurons surrounding the dendritic region of the mushroom bodies. Hypomorphic mutations of the essential dNR1 gene disrupt olfactory learning, and this learning defect is rescued with wild-type transgenes. Importantly, we show that Pavlovian learning is disrupted in adults within 15 hr after transient induction of a dNR1 antisense RNA transgene. Extended training is sufficient to overcome this initial learning defect, but long-term memory (LTM) specifically is abolished under these training conditions. CONCLUSIONS: Our study uses a combination of molecular-genetic tools to (1) generate genomic mutations of the dNR1 gene, (2) rescue the accompanying learning deficit with a dNR1+ transgene, and (3) rapidly and transiently knockdown dNR1+ expression in adults, thereby demonstrating an evolutionarily conserved role for the acute involvement of NMDARs in associative learning and memory. PMID- 15823534 TI - Distinct functions of the Tribolium zerknullt genes in serosa specification and dorsal closure. AB - BACKGROUND: In the long-germ insect Drosophila, a single extraembryonic membrane, the amnioserosa, covers the embryo at the dorsal side. In ancestral short-germ insects, an inner membrane, the amnion, covers the embryo ventrally, and an outer membrane, the serosa, completely surrounds the embryo. An early differentiation step partitions the uniform blastoderm into the anterior-dorsal serosa and the posterior-ventral germ rudiment giving rise to amnion and embryo proper. In Drosophila, amnioserosa formation depends on the dorsoventral patterning gene zerknullt (zen), a derived Hox3 gene. RESULTS: The short-germ beetle Tribolium castaneum possesses two zen homologs, Tc-zen1 and Tc-zen2. Tc-zen1 acts early and specifies the serosa. The loss of the serosa after Tc-zen1 RNAi is compensated by an expansion of the entire germ rudiment toward the anterior. Instead of the serosa, the amnion covers the embryo at the dorsal side, and later size regulation normalizes the early fate shifts, revealing a high degree of plasticity of short-germ development. Tc-zen2 acts later and initiates the amnion and serosa fusion required for dorsal closure. After Tc-zen2 RNAi, the amnion and serosa stay apart, and the embryo closes ventrally, assuming a completely everted (inside-out) topology. CONCLUSIONS: In Tribolium, the duplication of the zen genes was accompanied by subfunctionalization. One of the paralogues, Tc-zen1, acts as an early anterior-posterior patterning gene by specifying the serosa. In absence of the serosa, Tribolium embryogenesis acquires features of long-germ development with a single extraembryonic membrane. We discuss implications for the evolution of insect development including the origin of the zen-derived anterior determinant bicoid. PMID- 15823535 TI - Regulation of phytochrome B nuclear localization through light-dependent unmasking of nuclear-localization signals. AB - Phytochromes are red and far-red photoreceptors that regulate plant growth and development in response to environmental light cues. Phytochromes exist in two photo-interconvertible conformational states: an inactive Pr form and an active Pfr form. The alteration of phytochromes' subcellular location functions as a major regulatory mechanism of their biological activities. Whereas phytochromes in the Pr form localize in the cytoplasm, phytochromes in the Pfr form accumulate in the nucleus, where they interact with transcription factors to regulate gene expression. The molecular details of the regulation of phytochrome translocation by light are poorly understood. Using Arabidopsis phyB as a model, we demonstrate that the C-terminal PAS-related domain (PRD) is both necessary and sufficient for phyB nuclear import and that the entire C terminus is required for nuclear-body (NB) localization. We also show that phyB's N-terminal bilin lyase domain (BLD) and PHY domain interact directly with the PRD in a light-dependent manner. In vivo localization studies indicate that BLD-PHY is sufficient to regulate phyB's nuclear accumulation. For phyB nuclear localization, our results suggest a molecular mechanism in which the nuclear-localization signal in the PRD is masked by interactions with phyB's chromophore-attachment domains and unmasked by light dependent conformational changes. PMID- 15823536 TI - Cofactor-interaction motifs and the cooption of a homeotic Hox protein into the segmentation pathway of Drosophila melanogaster. AB - Some Drosophila Hox-complex members, including the segmentation gene fushi tarazu (Dm-ftz), have nonhomeotic functions. Characteristic expression in other arthropods supports an ancestral homeotic role for ftz, indicating that ftz function changed during arthropod evolution. Dm-Ftz segmentation function depends on interaction with ftz-F1 via an LXXLL motif and homeodomain N-terminal arm. Hox proteins interact with the cofactor Extradenticle (Exd) via their YPWM motif. Previously, we found that Dm-ftz mediates segmentation but not homeosis, whereas orthologs from grasshopper (Sg-ftz) and beetle (Tc-Ftz), both containing a YPWM motif, have homeotic function. Tc-Ftz, which unlike Sg-Ftz contains an LXXLL motif, displays stronger segmentation function than Sg-Ftz. Cofactor-interaction motifs were mutated in Dm-Ftz and Tc-Ftz and effects were evaluated in Drosophila to assess how these motifs contributed to Ftz evolution. Addition of YPWM to Dm Ftz confers weak homeotic function, which is increased by simultaneous LXXLL mutation. LXXLL is required for strong segmentation function, which is unimpeded by the YPWM, suggesting that acquisition of LXXLL specialized Ftz for segmentation. Strengthening the Ftz/Ftz-F1 interaction led to degeneration of the YPWM and loss of homeotic activity. Thus, small changes in protein sequence can result in a qualitative switch in function during evolution. PMID- 15823537 TI - The Drosophila homologue of the hereditary spastic paraplegia protein, spastin, severs and disassembles microtubules. AB - Hereditary spastic paraplegias (HSPs), a group of neurodegenerative disorders characterized by lower-extremity spasticity and weakness, are most commonly caused by mutations in the spastin gene, which encodes a AAA+ ATPase related to the microtubule-severing protein katanin. A Drosophila homolog of spastin (D spastin) was identified recently, and D-spastin RNAi-treated or genetic null flies show neurological defects, and protein overexpression decreases the density of cellular microtubules. Elucidating spastin's function and disease mechanism will require a more detailed understanding of its structure and biochemical mechanism. Here, we have investigated the effects of D-spastin, individual D spastin domains, and D-spastin proteins bearing disease mutations on microtubules in cellular and in vitro assays. We show that D-spastin, like katanin, displays ATPase activity and uses energy from ATP hydrolysis to sever and disassemble microtubules; disease mutations abolish or partially interfere with these activities. PMID- 15823538 TI - Phosphorylation of histone H4 serine 1 during DNA damage requires casein kinase II in S. cerevisiae. AB - Distinct patterns of posttranslational histone modifications can regulate DNA templated events such as mitosis, transcription, replication, apoptosis, and DNA damage, suggesting the presence of a "histone code" in these nuclear processes. Phosphorylation of histone H2A S129 at sites of DNA double-strand breaks (DSBs) has been implicated in damage repair in yeast. Here, we describe another phosphorylation event on serine 1 (S1) of histone H4; this event is also associated with MMS- or phleomycin-induced DSBs but not with UV-induced DNA damage. Chromatin-immunoprecipitation (ChIP) studies of an HO-endonuclease inducible strain show that S1 phosphorylation is specifically enhanced 20- to 25 fold in nucleosomes proximal to the DSB. In addition, we show that casein kinase II (CK2) can phosphorylate H4 S1 in vitro and that null or temperature-sensitive CK2 yeast mutants are defective for induction of H4 S1 phosphorylation upon DNA damage in vivo. Furthermore, H4 S1 phosphorylation and CK2 play a role in DSB re joining as indicated by a nonhomologous end-joining (NHEJ) plasmid assay. CK2 has been implicated in regulating a DNA-damage response; our data suggest that histone H4 S1 is one of its physiological substrates. These data suggest that this modification is a part of the DNA-repair histone code. PMID- 15823539 TI - Inhibition of a yeast LTR retrotransposon by human APOBEC3 cytidine deaminases. AB - The mammalian APOBEC3 family of cytidine deaminases includes several members that possess potent antiretroviral activity. Human APOBEC3F and APOBEC3G are specifically incorporated into human immunodeficiency virus type 1 (HIV-1) progeny virions in the absence of virion infectivity factor (Vif), where they deaminate deoxycytidine to deoxyuridine on the minus strand of nascent reverse transcripts. Editing of the HIV-1 cDNA leads to its degradation or to G to A hypermutation of the integrated provirus. Here, we show that APOBEC3 proteins also restrict the activity of a distantly related long terminal repeat (LTR) retrotransposon. When expressed in the yeast Saccharomyces cerevisiae, human APOBEC3C, APOBEC3F, or APOBEC3G or mouse APOBEC3 potently inhibit replication of the Ty1 LTR retrotransposon. APOBEC3G interacts with Ty1 Gag and is packaged into Ty1 virus-like particles (VLPs) by a mechanism that closely resembles the one it uses to enter HIV-1 virions. Expression of APOBEC3G results in a reduced level of Ty1 cDNA integration and G to A editing of integrated Ty1 cDNA. Our findings indicate that APOBEC3G restricts Ty1 and HIV-1 by similar mechanisms and suggest that the APOBEC3 proteins target a substantially broader spectrum of retroelements than previously appreciated. PMID- 15823540 TI - Defective skeletogenesis with kidney stone formation in dwarf zebrafish mutant for trpm7. AB - Development of the adult form requires coordinated growth and patterning of multiple traits in response to local gene activity as well as to global endocrine and physiological effectors. An excellent example of such coordination is the skeleton. Skeletal development depends on the differentiation and morphogenesis of multiple cell types to generate elements with distinct forms and functions throughout the body. We show that zebrafish touchtone/nutria mutants exhibit severe growth retardation and gross alterations in skeletal development in addition to embryonic melanophore and touch-response defects. These alterations include accelerated endochondral ossification but delayed intramembranous ossification, as well as skeletal deformities. We show that the touchtone/nutria phenotype results from mutations in trpm7, which encodes a transient receptor potential (TRP) family member that functions as both a cation channel and kinase. We find trpm7 expression in the mesonephric kidney and show that mutants develop kidney stones, indicating renal dysfunction. These results identify a requirement for trpm7 in growth and skeletogenesis and highlight the potential of forward genetic approaches to uncover physiological mechanisms contributing to the development of adult form. PMID- 15823541 TI - The meiotic defects of mutants in the Drosophila mps1 gene reveal a critical role of Mps1 in the segregation of achiasmate homologs. AB - The conserved kinase Mps1 is necessary for the proper functioning of the mitotic and meiotic spindle checkpoints (MSCs), which monitor the integrity of the spindle apparatus and prevent cells from progressing into anaphase until chromosomes are properly aligned on the metaphase plate. In Drosophila melanogaster, a null allele of the gene encoding Mps1 was recently shown to be required for the proper functioning of the MSC, but it did not appear to exhibit a defect in female meiosis. We demonstrate here that the meiotic mutant ald1 is a hypomorphic allele of the mps1 gene. Both ald1 and a P-insertion allele of mps1 exhibit defects in female meiotic chromosome segregation. The observed segregational defects are substantially more severe for pairs of achiasmate homologs, which are normally segregated by the achiasmate (or distributive) segregation system, than they are for chiasmate bivalents. Furthermore, cytological analysis of ald1 mutant oocytes reveals both a failure in the coorientation of achiasmate homologs at metaphase I and a defect in the maintenance of the chiasmate homolog associations that are normally observed at metaphase I. We conclude that Mps1 plays an important role in Drosophila female meiosis by regulating processes that are especially critical for ensuring the proper segregation of nonexchange chromosomes. PMID- 15823542 TI - Mitochondrial function and actin regulate dynamin-related protein 1-dependent mitochondrial fission. AB - Mitochondria display a variety of shapes, ranging from small and spherical or the classical tubular shape to extended networks. Shape transitions occur frequently and include fusion, fission, and branching. It was reported that some mitochondrial shape transitions are developmentally regulated, whereas others were linked to disease or apoptosis. However, if and how mitochondrial function controls mitochondrial shape through regulation of mitochondrial fission and fusion is unclear. Here, we show that inhibitors of electron transport, ATP synthase, or the permeability transition pore (mtPTP) induced reversible mitochondrial fission. Mitochondrial fission depended on dynamin-related protein 1 (DRP1) and F-actin: Disruption of F-actin attenuated fission and recruitment of DRP1 to mitochondria. In contrast, uncoupling of electron transport and oxidative phosphorylation caused mitochondria to adopt a distinct disk shape. This shape change was independent of the cytoskeleton and DRP1 and was most likely caused by swelling. Thus, disruption of mitochondrial function rapidly and reversibly altered mitochondrial shape either by activation of DRP1-dependent fission or by swelling, indicating a close relationship between mitochondrial fission, shape, and function. Furthermore, our results suggest that the actin cytoskeleton is involved in mitochondrial fission by facilitating mitochondrial recruitment of DRP1. PMID- 15823543 TI - Direct observation demonstrates that Liprin-alpha is required for trafficking of synaptic vesicles. AB - Axonal transport is required for the elaboration and maintenance of synaptic morphology and function. Liprin-alphas are scaffolding proteins important for synapse structure and electrophysiology. A reported interaction with Kinesin-3 (Kif1a) suggested Liprin-alpha may also be involved in axonal transport. Here, at the light and ultrastructural levels, we discover aberrant accumulations of synaptic vesicle markers (Synaptotagmin and Synaptobrevin-GFP) and clear-core vesicles along Drosophila Liprin-alpha mutant axons. Analysis of presynaptic markers reveals reduced levels at Liprin-alpha synapses. Direct visualization of Synaptobrevin-GFP transport in living animals demonstrates a decrease in anterograde processivity in Liprin-alpha mutants but also an increase in retrograde transport initiation. Pull-down assays reveal that Liprin-alpha interacts with Drosophila Kinesin-1 (Khc) but not dynein. Together, these findings suggest that Liprin-alpha promotes the delivery of synaptic material by a direct increase in kinesin processivity and an indirect suppression of dynein activation. This work is the first to use live observation in Drosophila mutants to demonstrate the role of a scaffolding protein in the regulation of bidirectional transport. It suggests the synaptic strength and morphology defects linked to Liprin-alpha may in part be due to a failure in the delivery of synaptic-vesicle precursors. PMID- 15823544 TI - Towards a resolution on the inherent methodological weakness of the "effective number of codons used by a gene". AB - Recently Anders Fuglsang provided a modified way for calculating N(c) when biased discrepancy is present in a gene [Biochem. Biophys. Res. Commun. 317 (2004) 957]. Instead of taking the average codon homozygosity for each synonymous family type (as proposed by Wright) [Gene 87 (1990) 23] Fuglsang considered codon homozygosity of each amino acid individually. Marsashi and Najafabadi [Biochem. Biophys. Res. Commun. 324 (2004) 1] in their recent article demonstrated that the readjustment for overestimation at the level of individual amino acids results in loss of considerable amount of information. Immediately after the publication of Marsashi and Najafabadi, Fuglsang proposed that codon homozygosities can be calculated based on the classical population genetics [Biochem. Biophys. Res. Commun. 327 (2005) 1]. Though Fuglsang's approach is a novel one, it fails when any of the amino acids are absent in a gene. However, the inherent cause of overestimation at the level of individual amino acids is still obscured in the literature. Here in this communication we have presented a general condition where effective number of codons is overestimated using Wright's formula and also we propose a new way to calculate N(c), which is independent of amino acid composition. PMID- 15823545 TI - A novel approach for over-expression, characterization, and isotopic enrichment of a homogeneous species of acyl carrier protein from Plasmodium falciparum. AB - Acyl carrier protein (ACP) plays a central role in fatty acid biosynthesis by transferring the acyl groups from one enzyme to another for the completion of the fatty acid synthesis cycle. Holo-ACP is the obligatory substrate for the synthesis of acyl-ACPs which act as the carrier and donor for various metabolic reactions. Despite its interactions with numerous proteins in the cell, its mode of interaction is poorly understood. Here, we report the over-expression of PfACP in minimal medium solely in its holo form and in high yield. Expression in minimal media provides a means to isotopically label PfACP for high resolution multi-nuclear and multi-dimensional NMR studies. Indeed, the proton-nitrogen correlated NMR spectrum exhibits very high chemical shift dispersion and resolution. We also show that holo-PfACP thus expressed is amenable to acylation reactions using Escherichia coli acyl-ACP synthetase as well as by standard chemical methods. PMID- 15823546 TI - Bm-TFF2, a trefoil factor protein with platelet activation activity from frog Bombina maxima skin secretions. AB - In mammals, trefoil factor family (TFF) proteins are involved in mucosal maintenance and repair, and they are also implicated in tumor suppression and cancer progression. A novel two domain TFF protein from frog Bombina maxima skin secretions (Bm-TFF2) has been purified and cloned. It activated human platelets in a dose-dependent manner and activation of integrin alpha(IIb)beta(3) was involved. Aspirin and apyrase did not largely reduce platelet response to Bm-TFF2 (a 30% inhibition), indicating that the aggregation is not substantially dependent on ADP and thromboxane A2 autocrine feedback. Elimination of external Ca(2+) with EGTA did not influence the platelet aggregation induced by Bm-TFF2, meanwhile a strong calcium signal (cytoplasmic Ca(2+) release) was detected, suggesting that activation of phospholipase C (PLC) is involved. Subsequent immunoblotting revealed that, unlike in platelets activated by stejnulxin (a glycoprotein VI agonist), PLCgamma2 was not phosphorylated in platelets activated by Bm-TFF2. FITC-labeled Bm-TFF2 bound to platelet membranes. Bm-TFF2 is the first TFF protein reported to possess human platelet activation activity. PMID- 15823547 TI - TNF-alpha promotes Doxorubicin-induced cell apoptosis and anti-cancer effect through downregulation of p21 in p53-deficient tumor cells. AB - p53 is a key regulator in cell apoptosis, and cancer cells deficient in p53 expression fail to respond to chemotherapy. Here we show that effective Doxorubicin (DOX)-induced apoptosis is p53-dependent. However, an alternative treatment of DOX/TNF-alpha/DOX restored sensitivity of p53-deficient cells to DOX induced apoptosis. Treatment of cells with TNF-alpha resulted in a decrease of p21 (waf1/cip1/sdi1) expression following second dose of DOX. In previous work, we demonstrated that p21 suppressed DOX-induced apoptosis via its (cyclin dependent kinase) CDK-binding and CDK-inhibitory activity. Thus, we propose that TNF-alpha enhances the anti-cancer effect of DOX through suppressing the anti apoptotic activity of p21, and that a combined treatment TNF-alpha/Dox is an effective chemotherapeutic strategy for p53-deficient cancers. PMID- 15823548 TI - Identification of a multiprotein "motor" complex binding to water channel aquaporin-2. AB - Targeted positioning of water channel aquaporin-2 (AQP2) strictly regulates body water homeostasis. Trafficking of AQP2 to the apical membrane is critical to the reabsorption of water in renal collecting ducts. Recently, we have identified for the first time proteins which directly bind to AQP2: SPA-1, a GTPase-activating protein for Rap1, and cytoskeletal protein actin. Based on these findings, we have speculated the existence of a multiprotein complex which includes AQP2, SPA 1, and actin, for providing the mechanism which generates force and motion in AQP2 trafficking. To clarify the proteins comprising the complex, a large amount of AQP2-associated protein complex was isolated from the extract of rat kidney papilla using immunoaffinity column coupled with anti-AQP2 antibody and was analyzed by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS). In addition to SPA-1 and actin, 11 proteins were identified using this method: ionized calcium binding adapter molecule 2, myosin regulatory light chain smooth muscle isoforms 2-A and 2-B, alpha-tropomyosin 5b, annexin A2 and A6, scinderin, gelsolin, alpha-actinin 4, alpha-II spectrin, and myosin heavy chain nonmuscle type A. Our findings show for the first time an AQP2 binding multiprotein "force generator" complex. This multiprotein complex may provide the machinery of driving AQP2 movement. PMID- 15823549 TI - Molecular components and toxicity of the venom of the solitary wasp, Anoplius samariensis. AB - The solitary spider wasp, Anoplius samariensis, is known to exhibit a unique long term, non-lethal paralysis in spiders that it uses as a food source for its larvae. However, neither detailed venom components nor paralytic compounds have ever been characterized. In this study, we examined the components in the low molecular weight fraction of the venom and the paralytic activity of the high molecular weight fraction. The major low molecular weight components of the venom were identified as gamma-aminobutyric acid and glutamic acid by micro-liquid chromatography/electrospray ionization mass spectrometry and nuclear magnetic resonance spectrometry analysis. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass analysis revealed that the A. samariensis venom contained the various proteins with weights of 4-100 kDa. A biological assay using Joro spiders (Nephila clavata) clearly showed that the high molecular weight fraction of the venom prepared by ultrafiltration exerted as potent non lethal long-term paralysis as the whole venom, whereas the low molecular weight fraction was devoid of any paralytic activity. These results indicated that several venomous proteins in the high molecular weight fraction are responsible for the paralytic activity. Furthermore, we determined the primary structure of one component designated As-fr-19, which was a novel multiple-cysteine peptide with high sequence similarity to several sea anemone and snake toxins including dendrotoxins, rather than any insect toxic peptides identified so far. Taken together, our data showed the unprecedented molecular and toxicological profiles of wasp venoms. PMID- 15823550 TI - The association of small heat shock protein Hsp16.3 with the plasma membrane of Mycobacterium tuberculosis: dissociation of oligomers is a prerequisite. AB - Hsp16.3, a small heat shock protein from Mycobacterium tuberculosis (MTB), was originally identified as an immuno-dominant antigen and later found to be a major membrane protein. In vitro studies show that Hsp16.3 exists as nonamers and undergoes dynamic dissociation/re-association equilibrium in solutions. Nevertheless, neither the details nor the physiological implications of the presence of Hsp16.3 in the plasma membrane have been studied. In this study, we demonstrated that the purified Hsp16.3 proteins were able to interact with the MTB plasma membrane in a specific and reversible manner, suggesting that there might be subunit exchange between membrane-bound Hsp16.3 and soluble Hsp16.3 oligomers. The dissociation of Hsp16.3 oligomers appears to be a prerequisite for its membrane binding, which is interesting in view that the dissociation of small heat shock protein oligomers was also found to be necessary for it to bind denaturing substrate proteins. Furthermore, the oligomeric structure of Hsp16.3 seems to be more dynamic and flexible when incubating with the mycobacterium lipids. The physiological implications of these observations for Hsp16.3, and small heat shock proteins in general, are discussed. PMID- 15823551 TI - Finding of endocannabinoids in human eye tissues: implications for glaucoma. AB - Cannabinoid CB(1) receptors are involved in ocular physiology and may regulate intraocular pressure (IOP). However, endocannabinoid levels in human ocular tissues of cornea, iris, ciliary body, retina, and choroid from normal and glaucomatous donors have not been investigated. Anandamide (N arachidonoylethanolamine; AEA), 2-arachidonoylglycerol (2-AG), and the anandamide congener, palmitoylethanolamide (PEA), were detected in all the human tissues examined. In eyes from patients with glaucoma, significantly decreased 2-AG and PEA levels were detected in the ciliary body, an important tissue in the regulation of IOP. The findings suggest that these endogenous compounds may have a role in this disease, particularly with respect to regulation of IOP. PMID- 15823552 TI - Identification and characterization of a novel human APH-1b splice variant lacking exon 4. AB - APH-1 is one of the four essential components of the presenilin-gamma-secretase complex and has two human homologs, APH-1a, and APH-1b, both of which are seven pass membrane proteins. Here, we identified a novel splice variant of human APH 1b. This variant lacks exon 4, which encodes the entire fourth transmembrane domain. The mRNA expression of this variant was detected in most tissues at low levels. In transiently transfected cells, protein expression of the APH-1b variant was much lower than that of the wild-type. Furthermore, exogenous expression of the APH-1-interacting protein, nicastrin, significantly increased the variant protein levels. These data suggest that the APH-1b variant protein is destabilized, and implies that the fourth transmembrane domain plays an important role in the protein stability and function of APH-1. PMID- 15823553 TI - 5-amino-imidazole carboxamide riboside acutely potentiates glucose-stimulated insulin secretion from mouse pancreatic islets by KATP channel-dependent and independent pathways. AB - AMP-activated protein kinase (AMPK) is an important signaling effector that couples cellular metabolism and function. The effects of AMPK activation on pancreatic beta-cell function remain unresolved. We used 5-amino-imidazole carboxamide riboside (AICAR), an activator of AMPK, to define the signaling mechanisms linking the activation of AMPK with insulin secretion. Application of 300 microM AICAR to mouse islets incubated in 5-14 mM glucose significantly increased AMPK activity and potentiated insulin secretion. AICAR inhibited ATP sensitive K(+) (K(ATP)) channels and increased the frequency of glucose-induced calcium oscillations in islets incubated in 8-14 mM glucose. At lower glucose concentration (5mM) AICAR did not affect K(ATP) activity or intracellular ([Ca(2+)](i)). AICAR also did not inhibit (86)Rb(+) efflux from islets isolated from Sur1(-/-) mice that lack K(ATP) channels yet significantly potentiated glucose stimulated insulin secretion. Our data suggest that AICAR stimulates insulin secretion by both K(ATP) channel-dependent and -independent pathways. PMID- 15823554 TI - Tumor necrosis factor-alpha induces differentiation of human peripheral blood mononuclear cells into osteoclasts through the induction of p21(WAF1/Cip1). AB - Tumor necrosis factor-alpha (TNF-alpha) is a multifunctional cytokine that mediates inflammation and induces bone loss caused by excessive bone resorption by osteoclasts. The interaction of TNF-alpha with its receptor activates several signal transduction pathways, including those of mitogen-activated protein (MAP) kinases (p38, JNK, and ERK) and NF-kappaB. Signaling from these molecules has been shown to play an important role in osteoclastogenesis. In the present study, we investigated the mechanism of TNF-alpha-induced osteoclast differentiation in human peripheral blood mononuclear cells (PBMCs). We found that TNF-alpha alone greatly induced differentiation of PBMCs into osteoclasts. The osteoclast differentiation induced by TNF-alpha was independent of RANKL binding to its receptor RANK on PBMCs. Furthermore, TNF-alpha potently activated p38 MAPK, JNK, and NF-kappaB. Western blotting analysis revealed that p21(WAF1/Cip1), a cyclin dependent kinase (CDK) inhibitor, is significantly induced upon TNF-alpha stimulation. The induction of p21(WAF1/Cip1) during differentiation is responsible for arrest at G(0)/G(1) phase and associated with the JNK pathway. These results suggest that TNF-alpha regulates osteoclast differentiation through p21(WAF1/Cip1) expression and further shows that these events require JNK activity. PMID- 15823555 TI - Trap RACK1 with Ras to mobilize Src signaling at syndecan-2/p120-GAP upon transformation with oncogenic ras. AB - HiTrap-syndecan-2/p120-GAP and HiTrap-syndecan-2/RACK1 affinity columns were applied to reveal that Src tyrosine kinase was highly expressed in BALB/3T3 cells transfected with plasmids pcDNA3.1-[S-ras(Q(61)K)] of shrimp Penaeus japonicus. Both columns were effective to isolate Src tyrosine kinase. The selective molecular affinity for Src was found to be stronger with HiTrap-syndecan-2/RACK1, as revealed with competitive RACK1 to dislodge Src from HiTrap-syndecan-2/p120 GAP. We thus challenged the syndecan-2/p120-GAP and syndecan-2/RACK1 with GTP K(B)-Ras(Q(61)K). The reaction between RACK1 and syndecan-2 was sustained in the presence of mutant Ras proteins, but not the reaction between p120-GAP and syndecan-2. In the presence of syndecan-2, GTP-K(B)-Ras(Q(61)K) exhibited sufficient reactivity with p120-GAP to discontinue the reaction between p120-GAP and syndecan-2. But the interference of mutant Ras disappeared when Src tyrosine kinase was introduced to stabilize the syndecan-2/p120-GAP complex. On the other hand, in the absence of syndecan-2, GTP-K(B)-Ras(Q(61)K) was found to react with RACK1. The reaction between GTP-K(B)-Ras(Q(61)K) and RACK1 could provide a mechanism to deprive RACK1 for the organization of syndecan-2/RACK1 complex and to facilitate the formation of syndecan-2/p120-GAP complex, as well as to provide docking sites for Src signaling upon transformation with oncogenic ras. PMID- 15823556 TI - Selenoprotein W as molecular target of methylmercury in human neuronal cells is down-regulated by GSH depletion. AB - Methylmercury (MeHg) is well known as a neurotoxic chemical. However, little is mentioned about its neurotoxic mechanism or molecular target in human neuronal cells in particular. We show in this study that exposure of human neuronal cell line, SH-SY5Y, to MeHg dose- and time-dependently impairs viability and mRNA expression of selenoprotein W (SeW) with a significant difference, unlike other selenoenzymes such as, SeP, GPX4, 5DI, and 5'DI. Using real-time RT PCR, the influence of selenium (Se) and glutathione (GSH) on SeW expression was also investigated. While Se depletion caused a weakly reduced SeW mRNA levels, additional Se caused an increase of SeW mRNA levels. Although 2 mM GSH had induced a weak shift on SeW level, the expression of SeW mRNA was down-regulated in SH-SY5Y cells treated with 25 microM BSO, an inhibitor of GSH synthesis. To understand the relationship between a decrease of SeW expression and intracellular GSH and ROS, we measured the concentration of intracellular GSH and ROS in cells treated to 1.4 microM MeHg using fluorescence based assays. A positive correlation was found between SeW mRNA level and intracellular GSH but no significant correlation was observed between intracellular ROS and SeW mRNA level or intracellular GSH contents. Therefore, we suggest that SeW is the novel molecular target of MeHg in human neuronal cells and down-regulation of this selenoenzyme by MeHg is dependent not on generation of ROS but on depletion of GSH. PMID- 15823557 TI - Neuronal differentiation of NTE-deficient embryonic stem cells. AB - Organophosphates induce neurological disorders. One of the enzymes inhibited by these compounds is neuropathy target esterase (NTE). In vitro, inhibition of NTE activity by organophosphates is correlated with inhibition of neurite initiation and reduction of neurite length, supporting the hypothesis that organophosphate induced neurological disorders are caused by inhibition of NTE activity. However, there is no direct evidence for the involvement of NTE in organophosphate-induced impairment of neurites in vitro. To examine the role of NTE, we have generated NTE-deficient mouse embryonic stem cells. These cells can differentiate into neuron-like cells. Although NTE-deficient cells exhibited a delay in neurite initiation in vitro, both the proportion of neuron-like cells which initiated neurites and the elongation of these neurites occurred at the normal rate. These results demonstrate that NTE activity is not required for neurite initiation or elongation per se, but is essential for the optimal rate of neurite initiation. PMID- 15823558 TI - Balanced GABAergic and glutamatergic synapse development in hippocampal neurons. AB - Coordinated development of excitatory and inhibitory synapses is crucial for normal function of neuronal circuits. Using homo- and heterochronic cultures of hippocampal neurons, we compared the formation of glutamatergic and GABAergic synapses at different stages and asked whether the age of dendrites affects their ability to accept new glutamatergic and GABAergic synapses. Neurons were transfected with either CFP-actin as a dendritic marker or GFP-synaptophysin as a presynaptic marker. We found that GFP-synaptophysin clusters formed on CFP-actin labeled dendrites at similar density regardless of pre- and postsynaptic cell type or the age of dendrites (0-2 weeks) upon co-culturing. Therefore, the age of mature dendrites does not affect their ability to accept new synapses. Because GABAergic transmission switches from depolarizing to hyperpolarizing during 1-2 weeks in these cultures, our observations also suggest that this developmental switch does not alter the formation of GABAergic synapses. PMID- 15823559 TI - Solution structure of BmP08, a novel short-chain scorpion toxin from Buthus martensi Karsch. AB - A novel short-chain scorpion toxin BmP08 was purified from the venom of the Chinese scorpion Buthus martensi Karsch by a combination of gel-filtration, ion exchange, and reversed-phase chromatography. The primary sequence of BmP08 was determined using the tandem MS/MS technique and Edman degradation, as well as results of NMR sequential assignments. It is composed of 31 amino acid residues including six cysteine residues and shares less than 25% sequence identity with the known alpha-KTx toxins. BmP08 shows no inhibitory activity on all tested voltage-dependent and Ca(2+)-activated potassium channels. The 3D-structure of BmP08 has been determined by 2D-NMR spectroscopy and molecular modeling techniques. This toxin adopts a common alpha/beta-motif, but shows a distinctive local conformation and features a 3(10)-helix and a shorter beta-sheet. The unique structure is closely related to the distinct primary sequence of the toxin, especially to the novel arrangement of S-S linkages in the molecule, in which two disulfide bridges (C(i)-C(j) and C(i+3)-C(j+3)) link covalently the 3(10)-helix with one strand of the beta-sheet structure. The electrostatic potential surface analysis of the toxin reveals salt bridges and hydrogen bonds between the basic residues and negatively charged residues nearby in BmP08, which may be unfavorable for its binding with the known voltage-dependent and Ca(2+) activated potassium channels. Thus, finding the target for this toxin should be an interesting task in the future. PMID- 15823560 TI - NELIN, a new F-actin associated protein, stimulates HeLa cell migration and adhesion. AB - A new gene (GenBank Accession No. AF114264) was cloned from umbilical vein wall tissue by using RT-PCR. The gene shares high similarity to the gene encoding F actin binding protein nexilin, so named as NELIN. A clone of 2737bp contains open reading frame of 1344bp extending from 412 to 1755. NELIN was expressed primarily in the heart and skeletal muscle among eight tested normal tissues. Immunofluorescence and immunoprecipitation demonstrated that NELIN product was associated with F-actin. Stable transfection of NELIN into HeLa cells increased the cell migration by 2.17-fold and the adhesion by 1.67-fold, respectively, compared to cells with the empty vector (P<0.05). The results support that NELIN product is an F-actin associated protein and mediates cell motility. PMID- 15823561 TI - Specific disruption of smooth muscle caldesmon expression in mice. AB - Caldesmon (CaD) is an actin-binding protein that is capable of inhibiting the actomyosin ATPase activity in vitro. CaD has a single gene that is alternatively spliced to generate the smooth muscle-specific form, h-CaD, and a shorter isoform, l-CaD, that is present only in non-muscle cells. The difference between h- and l-CaD is a highly charged repeating sequence, corresponding to a 35 nm long single helical region that separates the N-terminal domain from the C terminal domain of h-CaD. To test whether such an elongated h-CaD is essential for smooth muscles to function properly, we have specifically abrogated its expression in the mouse by targeting h-CaD without affecting the expression of l CaD. After genotyping, we have obtained homozygous knockout mice that indeed lack h-CaD, but nevertheless express varying amounts of l-CaD in a tissue-dependent fashion. The contractility of smooth muscles isolated from the knockout animals is currently under investigation. PMID- 15823562 TI - Genetic basis of phenotypic plasticity for predator-induced morphological defenses in anuran tadpole, Rana pirica, using cDNA subtraction and microarray analysis. AB - Anuran tadpoles (Rana pirica) are induced to develop a higher tail and a bulgy body as predator-specific morphological responses when they are exposed to predatory larval salamanders. Subtractive hybridization was performed using induced tadpole body skin and normal tadpoles' body skin. A total of 196 clones showed higher expression, and 104 clones showed lower expression, when they formed bulgy bodies. In the subtraction, carboxypeptidase B, trypsinogen, elastase I, fibrinogen, elastase II, triacyl-glycerol lipase, and alpha1 antitrypsin genes showed lower expression. In contrast, RT-like protein, bullous pemphigoid antigen, phosphoserine aminotransferase, uromodulin, tetranectin, chaperonin-like protein, zinc finger protein, osteonectin, aldehyde dehydrogenase, Sec 23A protein, and ribosomal protein showed higher gene expression. Microarray analysis was also performed using this subtracted cDNA (nine replicates). Results of the microarray data essentially corresponded with those of the subtraction data, and the degree of the suppressed genes was much stronger than that of the expressed genes. Carboxypeptidase B showed the strongest suppression, and its inhibition range was from 1/100 to 3/100 compared with that of control body skin. Strong suppression was also observed with trypsinogen, elastase I, fibrinogen, and elastase II as well. These results can be interpreted as increases of fibrinolysis by strong depression of both carboxypeptidase B and other genes simultaneously, resulting in the retention of blood vessels and facilitating the circulation of blood. Expression was observed in phosphoserine aminotransferase, aldehyde dehydrogenase, RT-related protein, chaperonin-like protein, tetranectin, bullous pemphigoid antigen, uromodulin, and Sec 23A protein. They were significantly (p<0.05) increased and were at least 1.5 times greater compared with the control. From the appearance, it seems that the bulgy shaped body is highly connecting to the bullous pemphigoid (BP) antigen that causes the skin blistering disorder, and tetranectin and uromodulin may be related to the extracell matrix through myogenesis, protein secretion, and ion transport, respectively. Since the RT-related protein gene derived from retrotransposon (L1) is known to disrupt mammalian transcriptomes, retrotransposon may be involved with phenotypic plasticity for morphological defense by Rana prica against predator threat. PMID- 15823564 TI - In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells. AB - In addition to long-term self-renewal capability, human mesenchymal stem cells (MSCs) possess versatile differentiation potential ranging from mesenchyme related multipotency to neuroectodermal and endodermal competency. Of particular concern is hepatogenic potential that can be used for liver-directed stem cell therapy and transplantation. In this study, we have investigated whether human umbilical cord blood (UCB)-derived MSCs are also able to differentiate into hepatocyte-like cells. MSCs isolated from UCB were cultured under the pro hepatogenic condition similar to that for bone marrow (BM)-derived MSCs. Expression of a variety of hepatic lineage markers was analyzed by flow cytometry, RT-PCR, Western blot, and immunofluorescence. The functionality of differentiated cells was assessed by their ability to incorporate DiI-acetylated low-density lipoprotein (DiI-Ac-LDL). As the cells were morphologically transformed into hepatocyte-like cells, they expressed Thy-1, c-Kit, and Flt-3 at the cell surface, as well as albumin, alpha-fetoprotein, and cytokeratin-18 and 19 in the interior. Moreover, about a half of the cells were found to acquire the capability to transport DiI-Ac-LDL. Based on these observations, and taking into account immense advantages of UCB over other stem cell sources, we conclude that UCB-derived MSCs retain hepatogenic potential suitable for cell therapy and transplantation against intractable liver diseases. PMID- 15823563 TI - Identification of MrgX2 as a human G-protein-coupled receptor for proadrenomedullin N-terminal peptides. AB - Proadrenomedullin N-terminal 20 peptide (PAMP[1-20]/PAMP-20) and its truncated analog, PAMP[9-20]/PAMP-12, are endogenous peptides that elicit hypotension through inhibiting catecholamine secretion from sympathetic nerve endings and adrenal chromaffin cells. Although the binding sites for PAMP are widely distributed, the nature of its receptor has been elusive. In an effort to identify potential PAMP receptor(s), we found that a human G-protein-coupled receptor, MrgX2, was specifically activated by PAMP. Although a previous study revealed that MrgX2 was a receptor for cortistatin, a neuropeptide involved in sleep regulation and locomotor activity, our present data indicated that the rank order of the agonistic effect against MrgX2 was "PAMP-12> or =cortistatin>PAMP 20". These activities were confirmed by the inhibition of the forskolin-elevated cAMP accumulation, Ca(2+) mobilization, and [(35)S]guanosine 5'-(gamma thio)triphosphate binding assays. These findings suggest that MrgX2 couples with not only G(alpha q) but also G(alpha i), consistent with previous reports on the pharmacological profile of PAMP signaling. Furthermore, by immunostaining, we found that MrgX2 was expressed in the adrenal chromaffin cells as well as the dorsal root ganglia. From these results, we concluded that MrgX2 is a potential human PAMP-12 receptor that regulates catecholamine secretion from adrenal glands. The present discovery will eventually lead to a better understanding of the pathophysiological role of proadrenomedullin peptides. PMID- 15823565 TI - Allele effects in MHC-peptide interactions: a theoretical analysis of HLA DRbeta1*0101-HA and HLA-DRbeta1*0401-HA complexes. AB - HLA-DRbeta1*0101-HA and HLA-DRbeta1*0401-HA complexes are studied and compared by means of their computationally derived multipolar moments and electrostatic potentials. Changes in electrostatic potential are associated with definite pocket interaction profiles. Thus, Pocket 1 projects itself as an anchoring pocket for both complexes, in accordance with experimental results. While Pocket 4 has an anchoring profile in the HLA-DRbeta1*0101 allele, it presents itself as modulating pocket-peptide interactions in HLA-DRbeta1*0401. Pockets 6 and 7 both strongly contribute to allele specificity, with Pocket 7 being very important for HLA-DRbeta1*0401-HA. Pocket 9 acts as a "double purpose" interaction site for both alleles. It both projects itself as an anchoring pocket as well as modulating pocket-peptide interactions. PMID- 15823566 TI - Synthesis of novel siRNAs having thymidine dimers consisting of a carbamate or a urea linkage at their 3' overhang regions and their ability to suppress human RNase L protein expression. AB - In order to examine the effect of modifications at the 3' overhang regions of short interfering RNAs (siRNAs) on their gene-silencing activities, we designed and synthesized novel siRNAs having thymidine dimers consisting of a carbamate or a urea linkage at their 3' overhang regions. Suppression of human RNase L protein expression by these siRNAs was analyzed by immunoblot with RNase L-specific antibody. It was found that, at 24 h post-transfection, the modified siRNAs having the thymidine dimers with the carbamate and urea linkage suppress the protein expression 78 and 37 times more efficiently than that with the natural phosphodiester linkage, respectively. Furthermore, the siRNA containing the carbamate linkage was 37 times more resistant to nucleolytic degradation by snake venom phosphodiesterase than the siRNA consisting of the natural phosphodiester linkage. Thus, the RNA duplexes having the thymidine dimers with the carbamate or urea linkage at their 3' overhang regions will be promising candidates for novel siRNA molecules to down-regulate protein expression. PMID- 15823567 TI - Elevation of Hook1 in a disease model of Batten disease does not affect a novel interaction between Ankyrin G and Hook1. AB - Hook1 is a member of a family of microtubule-binding proteins. Studies on the Drosophila homolog of Hook1 have suggested a role in the maturation and trafficking of internalized proteins to the late endosome. A weak interaction between Hook1 and the lysosomal/late endosomal protein, CLN3, was recently reported. Mutations in CLN3 result in the neurological disorder Batten disease. Here we show a novel interaction between Hook1 and Ankyrin G, an adaptor protein that binds the spectrin-actin cytoskeleton and targets proteins to the peripheral membrane. Although we demonstrate co-localization of Hook1 and Ankyrin G, Hook1 also localizes to additional regions of the cell devoid of Ankyrin G where it likely interacts with other proteins. There is no disruption of the Hook1-Ankyrin G interaction or localization in tissue derived from a Cln3-knockout mouse despite a nearly threefold increase in the expression of Hook1. However, mutation of CLN3 could lead to alterations in the functioning and positioning of organelles and membrane proteins through this Hook1-Ankyrin G interaction. PMID- 15823568 TI - Molecular insight into the effects of hypothyroidism on the developing cerebellum. AB - Despite the recognized importance of thyroid hormones for normal brain development, little is known about the critical molecular events underlying this role. We investigated the molecular basis of thyroid hormone action on the developing brain by comparing genome-wide gene expression patterns in the cerebellum between euthyroid and hypothyroid juvenile mice using microarrays. Pregnant dams were treated with 0.1% or 0.04% 6-propyl-2-thiouracil (PTU) in drinking water continuously from day 13 post conception until weaning to produce hypothyroid pups. Cerebella were collected from vehicle and 0.1% PTU treated pups at post-natal day (PND) 15, and mRNA from these was subjected to microarray analysis using Agilent high-density oligonucleotide chips. Statistical analysis (MAANOVA) revealed significant differential expression in 2940 genes including 1357 up- and 1583 down-regulated genes. Further analysis (combined MAANOVA and ANOVA) identified 204 significantly altered genes. Hypothyroidism had a greater effect on gene expression in male than in female pups. Transcriptional changes in several genes [Syt12 (Synaptotagmin 12), Rcor (RE1-silencing transcription factor co-repressor), Bag3 (Bcl-associated athanogene 3), p21, cyclin D, Bax (Bcl2 associated X protein), and Pcp2 (Purkinje cell protein 2)] were confirmed using real-time (RT) PCR analysis. Significantly altered expression of Bag3 in cerebella from PND 15 and PND 60 pups exposed to PTU suggests permanent functional alterations in the hypothyroid brain. The thyroid hormone negative regulation of Rcor expression was confirmed in vitro using HepG2 cells. In addition to Rcor, expression of several other genes that code for critical components of the REST (RE1-silencing transcription factor) pathway was shown to be altered in hypothyroid animals. These results suggest that modification of this pathway may have a significant role in causing impaired development in the hypothyroid brain. PMID- 15823569 TI - Differential impact of low temperature on fatty acid unsaturation and lipoxygenase activity in figleaf gourd and cucumber roots. AB - Previous studies show that low temperature strongly induces suberin layers in the roots of chilling-sensitive cucumber plants, while in contrast, low temperature produces a much weaker induction of suberin layers in the roots of the chilling tolerant figleaf gourd [S.H. Lee, G.C. Chung, S. Steudle, Gating of aquaporins by low temperature in roots of chilling-sensitive cucumber and -tolerant figleaf gourd, J. Exp. Bot. 56 (2005) 985-995; S.H. Lee, G.C. Chung, E. Steudle, Low temperature and mechanical stresses differently gate aquaporins of root cortical cells of chilling-sensitive cucumber and figleaf gourd, Plant Cell Environ. (2005) in press; S.J. Ahn, Y.J. Im, G.C. Chung, B.H. Cho, S.R. Suh, Physiological responses of grafted-cucumber leaves and rootstock roots affected by low root temperature, Scientia Hort. 81 (1999) 397-408]. Here, the effect of low temperature on fatty acid unsaturation and lipoxygenase activity was examined in cucumber and figleaf gourd. The double bond index demonstrated that membrane lipid unsaturation shows hyperbolic saturation curve in figleaf gourd roots while a biphasic response in cucumber roots to low temperature. In figleaf gourd, the hyperbolic response in the double bond index was primarily due to accumulation of linolenic acid. Chilling stress also significantly induced lipoxygenase activity in figleaf gourd roots. These results suggest that the degree of unsaturation of root plasma membrane lipids correlates positively with chilling-tolerance. Therefore, studies that compare the effects of chilling on cucumber and figleaf gourd may provide broad insight into stress response mechanisms in chilling sensitive and chilling-tolerant plants. Furthermore, these studies may provide important information regarding the relationship between lipid unsaturation and lipoxygenase function/activity, and between lipoxygenase activity and water channeling during the response to chilling stress. The possible roles of these processes in chilling tolerance are discussed. PMID- 15823570 TI - Escherichia coli outer membrane protein A adheres to human brain microvascular endothelial cells. AB - Escherichia coli K1 is the most common gram-negative bacterium causing neonatal meningitis. The outer membrane protein A (OmpA) assembles a beta-barrel structure having four surface-exposed loops in E. coli outer membrane. OmpA of meningitis causing E. coli K1 is shown to contribute to invasion of the human brain microvascular endothelial cells (HBMEC), the main cellular component of the blood brain barrier (BBB). However, the direct evidence of OmpA protein interacting with HBMEC is not clear. In this study, we showed that OmpA protein, solubilized from the outer membrane of E. coli, adhered to HBMEC surface. To verify OmpA interaction with the HBMEC, we purified N-terminal membrane-anchoring beta-barrel domain of OmpA and all surface-exposed loops deleted OmpA proteins, and showed that the surface-exposed loops of OmpA were responsible for adherence to HBMEC. These findings indicate that the OmpA is the adhesion molecule with HBMEC and the surface-exposed loops of OmpA are the determinant of this interaction. PMID- 15823571 TI - Homomeric and heteromeric interactions of the extracellular domains of death receptors and death decoy receptors. AB - Death receptors (DRs) can induce apoptosis by oligomerization with TRAIL, whereas death decoy receptors (DcRs) cannot, due to their lack of functional intracellular death domains. However, it is not known whether DRs and DcRs can interact with one another to form oligomeric complexes prior to TRAIL binding. To address this issue, the extracellular domains (ECDs) of DR4 (sDR4), DR5 (sDR5), DcR1 (sDcR1), and DcR2 (sDcR2) were expressed in a soluble, monomeric form, and their binding interactions were quantified by surface plasmon resonance. The purified sDRs and sDcRs exhibited native-like secondary structure and bound to TRAIL with binding affinities in the nanomolar range (K(D)= approximately 10-62 nM), suggesting that they were properly folded and functional. The soluble receptors interacted homophilically and heterophilically with similar micromolar range affinities (K(D)= approximately 1-9 microM), with the exception that sDR5 did not interact with the sDcRs. Our results suggest that most DRs and DcRs can laterally interact through their ECDs to form homomeric and/or heteromeric complexes in the absence of TRAIL binding. PMID- 15823572 TI - A stable FeIII-FeIV replacement of tyrosyl radical in a class I ribonucleotide reductase. AB - Ribonucleotide reductase (RNR) of Chlamydia trachomatis is a class I RNR enzyme composed of two homodimeric components, proteins R1 and R2. In class I RNR, R1 has the substrate binding site, whereas R2 has a diferric site and normally in its active form a stable tyrosyl free radical. C. trachomatis RNR is unusual, because its R2 component has a phenylalanine in the place of the radical carrier tyrosine. Replacing the tyrosyl radical, a paramagnetic Fe(III)-Fe(IV) species (species X, normally a transient intermediate in the process leading to radical formation) may provide the oxidation equivalent needed to start the catalytic process via long range electron transfer from the active site in R1. Here EPR spectroscopy shows that in C. trachomatis RNR, species X can become essentially stable when formed in a complete RNR (R1/R2/substrate) complex, adding further weight to the possible role of this species X in the catalytic reaction. PMID- 15823573 TI - Suppression of hepatitis A virus genome translation and replication by siRNAs targeting the internal ribosomal entry site. AB - Small interfering RNAs (siRNAs) targeting the coding region of hepatitis A virus (HAV) were shown to specifically inhibit viral genome replication. Compared to the coding region, the HAV internal ribosomal entry site (IRES) in the 5' non coding region is highly sequence-conserved and folds into stable secondary structures. Here, we report efficient and sustained RNA interference mediated by both RNase III-prepared siRNA (esiRNA) and vector-derived short hairpin RNAs (shRNAs) that are targeted to various domains of the HAV IRES. Using reporter constructs, and the DNA-based HAV replicon system, we found that shRNAs targeting the HAV IRES domains IIIc and V sustainably suppressed genome translation and replication whereas the IRES domains IIIa and IV were resistant to RNA interference. Our study suggests that some HAV IRES domains might be used as a universal and effective target for specific inhibition of HAV infection. PMID- 15823574 TI - Brain PP2A is modified by thiol-disulfide exchange and intermolecular disulfide formation. AB - The regulation of protein phosphatase 2A (PP2A) activity by thiol-disulfide exchange and resulting formation of an intermolecular disulfide was examined following exposure of a rat brain soluble fraction to a biotinylated derivative of the model disulfide HPDP (HPDP-biotin) which would be expected to label reactive protein thiols with a disulfide-linked biotin. The results show that a low concentration (500 microM) of HPDP-biotin produced substantial inhibition of PP2A activity and promoted the binding of the catalytic subunit of PP2A to an immobilized avidin-affinity column. Both the inhibition of PP2A activity and the binding of PP2A to the avidin column were reversed by treatment with the disulfide reducing agent dithiothreitol (DTT). Furthermore, the specific activity of PP2A was up to 7-fold higher in the DTT-eluted fractions from the avidin affinity column than in the soluble fraction. These findings demonstrate directly that PP2A is susceptible to reversible inhibitory modification by thiol-disulfide exchange and provide mechanistic support for the emerging view that PP2A is an oxidant-sensitive protein phosphatase. PMID- 15823576 TI - Competitive inhibition of the dengue virus NS3 serine protease by synthetic peptides representing polyprotein cleavage sites. AB - The NS3 serine protease of dengue virus is required for the maturation of the viral polyprotein and consequently represents a promising target for the development of antiviral inhibitors. However, the substrate specificity of this enzyme has been characterized only to a limited extent. In this study, we have investigated product inhibition of the NS3 protease by synthetic peptides derived from the P6-P1 and the P1'-P5' regions of the natural polyprotein substrate. N terminal cleavage site peptides corresponding to the P6-P1 region of the polyprotein were found to act as competitive inhibitors of the enzyme with K(i) values ranging from 67 to 12 microM. The lowest K(i) value was found for the peptide representing the NS2A/NS2B cleavage site, RTSKKR. Inhibition by this cleavage site sequence was analyzed by using shorter peptides, SKKR, KKR, KR, AGRR, and GKR. With the exception of the peptide AGRR which did not inhibit the protease at a concentration of 1mM, all other peptides displayed K(i) values in the range from 188 to 22 microM. Peptides corresponding to the P1'-P5' region of the polyprotein cleavage sites had no effect on enzymatic activity at a concentration of 1mM. Molecular docking data of peptide inhibitors to a homology based model of the dengue virus type 2 NS2B(H)-NS3p co-complex indicate that binding of the non-prime site product inhibitors is similar to ground-state binding of the corresponding substrates. PMID- 15823575 TI - Transiently truncated and differentially regulated expression of midkine during mouse embryogenesis. AB - Midkine (MK) is a retinoic acid response cytokine, mostly expressed in embryonic tissues. Aberrant expression of MK was found in numerous cancers. In human, a truncated MK was expressed specifically in tumor/cancer tissues. Here we report the discovery of a novel truncated form of MK transiently expressed during normal mouse embryonic development. In addition, MK is concentrated at the interface between developing epithelium and mesenchyme as well as highly proliferating cells. Its expression, which is closely coordinated with angiogenesis and vasculogenesis, is spatiotemporally regulated with peaks in extensive organogenesis period and undifferentiated cells tailing off in maturing cells, implying its role in nascent blood vessel (endothelial) signaling of tissue differentiation and stem cell renewal/differentiation. Cloning and sequencing analysis revealed that the embryonic truncated MK, in which the conserved domain is in-frame deleted, presumably producing a novel secreted small peptide, is different from the truncated form in human cancer tissues, whose deletion results in a frame-shift mutation. Our data suggest that MK may play a role in epithelium mesenchyme interactions, blood vessel signaling, and the decision of proliferation vs differentiation. Detection of the transiently expressed truncated MK reveals its novel function in development and sheds light on its role in carcinogenesis. PMID- 15823577 TI - Alternative splicing of the human Kank gene produces two types of Kank protein. AB - The human Kank gene encodes an ankyrin repeat domain-containing protein which regulates actin polymerization. There are at least two types of Kank protein depending on cell type, likely due to differences in transcription. Here, to examine the transcriptional initiation and genomic organization of the human Kank gene, we performed 5'-RACE (rapid amplification of cDNA ends) using total RNA from normal kidney and a human kidney cancer cell line, VMRC-RCW cells. The results suggest that the human Kank gene has several alternative first exons. While mRNA from VMRC-RCW cells encoded Kank protein (referred to as Kank-S) as reported previously, mRNA from the normal kidney tissue encoded a novel type of Kank protein (referred to as Kank-L), which contained an additional N-terminal sequence 158 amino acids long. Promoter activity and the expression of the Kank variants in normal and cancer tissues were examined. PMID- 15823578 TI - Binding affinity difference induced by the stereochemistry of the sulfoxide bridge of the cyclic peptide inhibitors of Grb2-SH2 domain: NMR studies for the structural origin. AB - The SAR study on a phage library-derived non-phosphorylated cyclic peptide ligand of Grb2-SH2 domain indicates that the configuration of the cyclization linkage is crucial for assuming the active binding conformation. When the thioether linkage was oxidized to the two chiral sulfoxides, the R-configured sulfoxide-cyclized peptide displayed 10-30 times more potency than the corresponding S-configured one in binding affinity to the Grb2-SH2 domain. In this paper, the solution structures of such a pair of sulfoxide-bridged cyclic peptide diastereoisomers, i.e., cyclo[CH(2)CO-Gla(1)-L-Y-E-N-V-G-NPG-Y-(R/S)C(O)(10)]-amide, were determined by NMR and molecular dynamics simulation. Results indicate that the consensus sequence of Y(3)-E(4)-N(5)-V(6) in both diastereoisomers adopt a beta turn conformation; however, the R-configured peptide forms an extended structure with a circular backbone conformation, while the S-configured isomer forms a compact structure with key residues buried inside the molecule. The average root mean-square deviations were found to be 0.756 and 0.804 A, respectively. It is apparent that the chiral S-->O group played a key role in the solution structures of the sulfoxide-bridged cyclic peptides. The R-sulfoxide group forms an intramolecular hydrogen bond with the C-terminal amide, conferring a more rigid conformation with all residues protruding outside except for Leu2, in which the Gla1 and Tyr3 share an overlapping function as previous SAR studies proposed. Additionally, the extended structure endows a more hydrophilic binding surface of the R-configured peptide to facilitate its capture by its targeted protein. In comparison, the S-configured sulfoxide was embedded inside the ligand peptide leading to a compact structure, in which the essential residues of Gla1, Tyr3, and Asn5 form multiple intramolecular hydrogen bonds resulting in an unfavorable conformational change and a substantial loss of the interaction with the protein. The solution structures disclosed by our NMR and molecular dynamics simulation studies provide a molecular basis for understanding how the chirality of the cyclization linkage remarkably discriminates in terms of the binding affinity, thus advancing the rational design of potent non-phosphorylated inhibitors of Grb2-SH2 domain as antitumor agents. PMID- 15823579 TI - Functional dissection of HCMV US11 in mediating the degradation of MHC class I molecules. AB - The human cytomegalovirus (HCMV) gene product US11 dislocates MHC I heavy chains from the endoplasmic reticulum (ER) and targets them for proteasomal degradation in the cytosol. To identify the structural and functional domains of US11 that mediate MHC class I molecule degradation, we constructed truncated mutants and chimeric proteins, and analyzed these to determine their intracellular localization and their ability to degrade MHC class I molecules. We found that only the luminal domain of US11 was essential to confer ER localization to the protein but that the ability to degrade MHC class I molecules required both the transmembrane domain and the luminal domain of US11. By analyzing a series of point mutants of the transmembrane domain, we were also able to identify Gln(192) and Gly(196) as being crucial for the functioning of US11, suggesting that these residues may play a critical role in interacting with the components of the protein degradation machinery. PMID- 15823580 TI - Antiangiogenic activity of 4-O-methylgallic acid from Canavalia gladiata, a dietary legume. AB - Development of nontoxic and biologically safe antiangiogenic agent has been highlighted as a promising way to treat angiogenesis related diseases including cancer. Herein, we isolated 4-O-methylgallic acid (4-OMGA) from the seed of Canavalia gladiata, a dietary legume, on the basis of the growth inhibitory activity for bovine aortic endothelial cells (BAECs). The compound potently inhibits endothelial cell invasion and tube formation stimulated with basic fibroblast growth factor (bFGF) at low micromolar concentrations where it shows no cytotoxicity to the cells. In addition, 4-OMGA inhibits vascular endothelial cell growth factor (VEGF) production under hypoxic condition and the production of reactive oxygen species (ROS) in the endothelial cells stimulated with VEGF. These results demonstrate that 4-OMGA is a compound having potential for an antiangiogenic agent. PMID- 15823581 TI - Macrophage colony-stimulating factor expression in retrovirally transduced cells is dependent upon both the adherence status of the target cells and its 5' flanking untranslated region. AB - Numerous cell types retrovirally transduced with macrophage colony-stimulating factor (M-CSF) using LXSN-based vectors showed a variable expression of the transgene. Expression of M-CSF correlated with the cells' adherent status. Transduced adherent cells produced the M-CSF, whereas the non-adherent cells synthesized little M-CSF. Studies showed that the 5'-UTR of the M-CSF gene regulated transgenic M-CSF gene expression. Ligation of this 5'-UTR to the enhanced green fluorescent protein gene (EGFP) caused the expression of EGFP to show the same dichotomy as previously seen with the M-CSF. Transgenic M-CSF was expressed within non-adherent cells when the 5'-UTR was removed from the LXSN vector. Quantitative real-time polymerase chain reaction analysis confirmed that lesser production of M-CSF mRNA occurred within the non-adherent cells than in the adherent cells. This difference was eliminated when the 5'-UTR was removed from the retroviral vector. Our work suggests that this 5'-UTR of the M-CSF gene could be an important way to get transgenic expression within adherent cells, but not in non-adherent cells. PMID- 15823582 TI - Stimulation of nicotinic acetylcholine receptors protects motor neurons. AB - The present study demonstrated that administration of nicotine prevented glutamate-induced motor neuronal death in primary cultures of the rat spinal cord. The nicotine-induced neuroprotection was inhibited by either dihydro-beta erythroidin (DHbetaE) or alpha-bungarotoxin (alphaBT), suggesting that it is mediated through both alpha4beta2 and alpha7 nicotinic acetylcholine receptors (nAChRs). Both alpha4beta2 and alpha7 nAChRs were identified on rat spinal motor neurons by immunohistochemical methods. We also demonstrated that galantamine, an acetylcholinesterase inhibitor with allosteric nAChR-potentiating ligand properties, prevented glutamate-induced motor neuronal death. These results suggest that stimulation of nAChR may be used as a treatment for ALS. PMID- 15823583 TI - ADP-ribosyl cyclase couples to cyclic AMP signaling in the cardiomyocytes. AB - ADP-ribosyl cyclase (ADPR-cyclase) produces a Ca(2+)-mobilizing second messenger cyclic ADP-ribose (cADPR) from beta-NAD(+). In this study, we examined the molecular basis of which beta-adrenergic receptor (betaAR) stimulation induces cADPR formation and characterized cardiac ADPR-cyclase. The results revealed that isoproterenol-mediated increase of [Ca(2+)](i) in rat cardiomyocytes was blocked by pretreatment with a cADPR antagonistic derivative 8-Br-cADPR, a PKA inhibitor H89 or high concentration of ryanodine. Moreover, incubation of ventricular lysates with isoproterenol, forskolin or cAMP resulted in activation of ADPR cyclase that was inhibited by pretreatment with H89. Supporting the observations, the cADPR antagonist and H89 blocked 8-CPT-cAMP, a cell-permeant cAMP analog induced increase in [Ca(2+)](i) but not cGMP-mediated increase. Characterization of partially purified cardiac ADPR-cyclase showed a molecular mass of approximately 42 kDa and no cross-activity with CD38 antibodies, and the enzyme activity was inhibited by Zn(2+) but not dithiothreitol. Microinjection of the enzyme into rat cardiomyocytes increased the level of [Ca(2+)](i) in a concentration-dependent manner. The enzyme-mediated increase of [Ca(2+)](i) was blocked by the cADPR antagonist. These findings suggest that betaAR-mediated regulation of [Ca(2+)](i) in rat cardiomyocytes is primed by activation of cardiac ADPR-cyclase via cAMP/PKA signaling and that cardiac ADPR-cyclase differs from CD38 in biochemical and immunological properties. PMID- 15823584 TI - In vitro trans-differentiation of rat mesenchymal cells into insulin-producing cells by rat pancreatic extract. AB - Recent reports have suggested that mesenchymal cells derived from bone marrow may differentiate into not only mesenchymal lineage cells but also other lineage cells. There is possibility for insulin-producing cells (IPCs) to be differentiated from mesenchymal cells. We used self-functional repair stimuli of stem cells by partial injury. Rat pancreatic extract (RPE) from the regenerating pancreas (2 days after 60% pancreatectomy) was treated to rat mesenchymal cells. After the treatment of RPE, they made clusters like islet of Langerhans within a week and expressed four pancreatic endocrine hormones; insulin, glucagon, pancreatic polypeptide, and somatostatin. Moreover, IPCs released insulin in response to normal glucose challenge. Here we demonstrate that the treatment of RPE can differentiate rat mesenchymal cells into IPCs which can be a potential source for the therapy of diabetes. PMID- 15823585 TI - Suppression of tumor cell invasiveness by hydrolyzable tannins (plant polyphenols) via the inhibition of matrix metalloproteinase-2/-9 activity. AB - Elevated expression of matrix metalloproteinases (MMPs), especially that of MMP-2 and MMP-9, is associated with increased metastatic potential in many tumor cells. Recently, green tea polyphenol epigallocatechin-3-O-gallate (EGCG) has been shown to inhibit the MMP-2/-9 activity as well as the invasiveness of tumor cells. In this study, we have examined the inhibitory effect of hydrolyzable tannins (plant polyphenols) on the tumor cell invasion. Our results demonstrate that beta-d glucose whose hydroxy groups are substituted entirely with galloyl group and further some of them are cross-linked to form hexahydroxydiphenoyl group, for example, suppresses the invasiveness of tumor cells much more potently than EGCG via direct inhibition of the MMP-2/-9 activity. Among those examined, 1,2,4-tri-O galloyl-3,6-hexahydroxydiphenoyl-beta-d-glucose (punicafolin) inhibits the invasion of HT1080 fibrosarcoma cells most potently. These hydrolyzable tannins would provide new leads for the development of potent inhibitors against tumor metastasis. PMID- 15823586 TI - PKCalpha is involved in phorbol ester TPA-mediated stabilization of p14ARF. AB - We generated A21-13 cells expressing p14(ARF) in the presence of doxycycline in order to examine the stability of p14(ARF) protein. The effects of proteasome inhibitor MG132 on p14(ARF) protein stabilization were detectable using our experimental procedure. Introduction of mutant p53 did not affect MG132-mediated p14(ARF) protein stabilization. We found that phorbol ester TPA (12-o tetradecanoyl-phorbol 13-acetate) stabilized p14(ARF) protein and that p53 status had no effect on TPA-mediated stabilization. TPA-mediated stabilization was abolished by staurosporine but not by lovastatin or U0126. We further investigated which isoforms of PKC were involved in TPA-mediated p14(ARF) stabilization using short-interference RNA. Knockdown of PKCalpha, but not PKCdelta, attenuated TPA-mediated p14(ARF) stabilization. These findings suggest that PKCalpha is involved in TPA-mediated stabilization of p14(ARF) protein, and this effect of TPA was not affected by the Ras/MAPK pathway or p53 status. Our results are indicative of a novel role of PKC in p14(ARF) protein stability. PMID- 15823588 TI - Introduction to bioengineering: melding of engineering and biological sciences. AB - Engineering has traditionally focused on the external extensions of organisms, such as transportation systems, high-rise buildings, and entertainment systems. In contrast, bioengineering is concerned with inward processes of biologic organisms. Utilization of engineering principles and techniques in the analysis and solution of problems in medicine and biology is the basis for bioengineering. This article discusses subspecialties in bioengineering and presents examples of projects in this discipline. PMID- 15823589 TI - Cell-based therapies and tissue engineering. AB - Tissue engineering is a rapidly evolving discipline that may some-day afford surgeons a limitless supply of autologous tissue for transplantation or allow in situ tissue regeneration. A number of biologic, engineering, and clinical challenges continue to face tissue engineers and surgeons alike. One important example is the choice of an appropriate cell scaffold that promotes growth and is eventually resorbed by the body. Although the application of bioengineered tissue is specific to the anatomic areas of interest,continued advances bring tissue engineering closer to reality in all areas of otolaryngology. PMID- 15823590 TI - Biophotonic and other physical methods for characterizing oral mucosa. AB - This article discusses biophotonic and other physical methods for characterizing oral mucosa. PMID- 15823591 TI - Selected laser-based therapies in otolaryngology. AB - This article has discussed selected applications of laser therapies in otolaryngology. Lasers are the treatment of choice for PWS lesions, but new advances are needed to achieve complete blanching in the majority of cases. Noninvasive in vivo imaging and laser therapy coupled with PDT are two areas showing promise to improve outcome. PDT has the potential to offer new, cost effective treatment options, and possibly cure, to cancer patients who are not candidates for traditional surgery, chemotherapy, and radiotherapy. PMID- 15823593 TI - Nanotechnology in otolaryngology. AB - This article provides a background on nanotechnology and discusses the applications of nanotechnology to medicine, specifically to otolaryngology. PMID- 15823592 TI - Bioengineering solutions for hearing loss and related disorders. AB - Advances in digital signal processing, microelectronics, and power technology have produced devices that have contributed significantly to the quality of life and communication abilities of individuals with hearing impairment and tinnitus. Future technological developments will expand the benefits of current devices and offer new treatments for otologic disorders. PMID- 15823594 TI - Progress towards seamless tissue fusion for wound closure. AB - Tissue fusion shows great promise in creating the ideal wound closure;however devices and materials are still at an early stage of development.Energy-based closure methods, such as laser tissue welding, have proven that a thermal mediated tissue fusion can result in a closure that is physiologically and mechanically seamless, and has sufficient tensile strength.However, the techniques are not easily reproducible and are not cost effective, and therefore they are not gaining wide acceptance. Nevertheless,the work of the scientists who have been exploring tissue welding has laid the foundation for more rapid development of new systems that can deliver energy more efficiently and with greater control. Some additional energy-based systems are available or are being developed that show great promise;however, clinical efficacy has yet to be demonstrated. PMID- 15823595 TI - Advances in head and neck imaging. AB - This article discusses new techniques in head and neck imaging. PMID- 15823596 TI - Bioinformatics in otolaryngology. AB - This article reviews the recent literature on microarray analysis, bioinformatics techniques, and genomics in relation to the study of carcinogenesis of head and neck cancers. PMID- 15823597 TI - Distraction osteogenesis in the craniofacial skeleton. AB - Bone deficiency in the craniofacial skeleton has posed a challenge to craniomaxillofacial surgeons for many decades. Sliding osteotomies and bone grafting have been traditional staples for correcting bone deficiency. In recent years, especially with the advent of newer devices, mechanically induced growth of bone and soft tissue has be-come a major tool in the correction of bony deficiency of the skull and face. In this article, the authors discuss the physiology and technique of distraction osteogenesis, or mechanically induced growth,for the correction of congenital and acquired facial deformities. PMID- 15823598 TI - Soft tissue implants and fillers. AB - The number and variety of soft tissue implants and fillers at the clinician's disposal continues to increase rapidly as new materials and bioengineering techniques evolve. The ideal implant for all purposes does not yet exist, so the clinician must be familiar with a wide variety of products for each application. Continued advances in synthetic, allogeneic, and autologous products and experience with existing products will further augment the surgeon's armamentarium in the coming years. PMID- 15823599 TI - New radiation therapy techniques for the treatment of head and neck cancer. AB - This article reviews the most recent technology used in the treatment of head and neck cancer. It discusses brachytherapy, new ways to mix radionuclides for enhanced radiobiologic effects, and different fractionation schemes that have grown in clinical importance. Intensity-modulated radiotherapy has become a mainstay in head and neck cancer treatment, and the authors discuss several popular and emerging approaches. Patient immobilization and imaging are also discussed. PMID- 15823600 TI - Alternative surgical dissection techniques. AB - The bipolar scissors, coblator, harmonic scalpel, and somnoplasty techniques are widely available and offer new choices for the operating arena. There are advantages and disadvantages to all four techniques. With time, these dissection methods will prove their lasting power. Otolaryngologists have already begun to expand their applications and will surely play a role in their use and development. PMID- 15823601 TI - Cooperative interaction of p65 and C/EBPbeta modulates transcription of BKV early promoter. AB - Reactivation of the human polyomavirus BK (BKV) has emerged as an important cause of allograft rejection in renal transplant recipients. Expression of the viral early promoter that leads to production of T-antigen is the first event in viral lytic infection. In an effort to understand the mechanism involved in the activation of BKV early gene (BKV(E)) expression, we analyzed the promoter/enhancer region of the virus and identified binding motifs for the inducible transcription factors NF-kappaB and C/EBPbeta, which are in juxtaposition to each other downstream from the early gene transcription initiation site. Results from transfection studies demonstrate that overexpression of the p65 subunit of NF-kappaB, but not C/EBPbeta stimulates transcription of the BKV(E) promoter in CV-1 cells. Interestingly, low level expression of C/EBPbeta showed a synergistic effect on p65 activation of the BKV(E) promoter, suggesting a functional cooperativity between these two regulators upon viral gene transcription. Results from DNA-binding studies showed the ability of p65 and C/EBPbeta to bind independently with BKV DNA as removal of the binding site for p65 or C/EBPbeta had no significant effect on the interaction of p65 and C/EBPbeta with their motifs, respectively. Functional evaluation of the mutant promoter with no binding sites for either NF-kappaB or C/EBPbeta showed that the observed synergism requires the p65 but not the C/EBPbeta binding site, suggesting cross-talk between C/EBPbeta and p65 in this event. Results from the co-expression of p65 and C/EBPbeta showed no evidence for the formation of a DNA-protein complex containing both p65 and C/EBPbeta, although results from protein-protein interaction studies verified the ability of C/EBPbeta to interact with p65. A dominant-negative isoform of C/EBPbeta which contains the DNA binding but not activation domain of full-length C/EBPbeta cooperated with p65 in activating the BKV(E) promoter, suggesting a functional interaction between the b-ZIP domain of C/EBPbeta and NF-kappaB. PMID- 15823602 TI - Serotype-specific replicating AAV helper constructs increase recombinant AAV type 2 vector production. AB - One of the major limitations of the use of adeno-associated virus (AAV) as a tool for gene therapy is the difficulty in providing sufficient quantities of the virus for pre-clinical and clinical trials. Here, we report a novel approach for amplifying AAV helper functions, which mimics the normal replication of wild type (wt) AAV resulting in a high yield of AAV vectors. Cotransfection of replicating but non-packaging AAV helper constructs in the presence of adenovirus (Ad) produces a high level of Rep and Cap proteins. Yield of AAV2/GFP vector obtained from this helper DNA replication system was approximately 20-fold higher than traditional methods. Molecular analysis suggested that virus yield was associated with capsid protein production. The transfection ratio was optimized using these novel helper constructs, resulting in an additional 2-fold increase in vector yield without presence of replication competent AAV (rcAAV). This strategy supports development of AAV packaging systems that retain normal virus replication capability without helper virus encapsidation. PMID- 15823603 TI - SH1: A novel, spherical halovirus isolated from an Australian hypersaline lake. AB - A novel halovirus, SH1, with a spherical morphology is described. Isolated from a hypersaline lake, SH1 is divalent, producing clear plaques on Haloarcula hispanica and a natural Halorubrum isolate. Single-step growth curves gave a latent period of 5-6 h and a burst size of around 200 PFU/cell. The host can differentiate to form tight clusters of thick cell-walled forms, and these were shown to be resistant to infection. Purified virions had no visible tail, were about 70 nm in diameter, and displayed a fragile outer capsid layer, possibly with an underlying membrane component. The structural proteins of the virion were analyzed by SDS-PAGE and several were found to be cross-linked, forming protein complexes. The genome was linear, dsDNA, of approximately 30 kb in length. This morphology and linear genome are features not observed in any other euryarchaeal viruses, but have properties similar to the bacterial virus PRD1. PMID- 15823604 TI - Identification of murine CD8 T cell epitopes in codon-optimized SARS-associated coronavirus spike protein. AB - The causative agent of severe acute respiratory syndrome (SARS) has been identified as a new type of coronavirus, SARS-associated coronavirus (SARS-CoV). CD8 T cells play an important role in controlling diseases caused by other coronaviruses and in mediating vaccine-induced protective immunity in corresponding animal models. The spike protein, a main surface antigen of SARS CoV, is one of the most important antigen candidates for vaccine design. Overlapping peptides were used to identify major histocompatibility complex class I-restricted epitopes in mice immunized with vectors encoding codon-optimized SARS-CoV spike protein. CD8 T-cell responses were mapped to two H-2(b)-restricted epitopes (S436-443 and S525-532) and one H-2(d)-restricted epitope (S366-374). The identification of these epitopes will facilitate the evaluation of vaccine strategies in murine models of SARS-CoV infection. Furthermore, codon and promoter optimizations can greatly enhance the overall immunogenicity of spike protein in the context of replication-defective human and simian adenoviral vaccine carriers. The optimized recombinant adenoviral vaccine vectors encoding spike can generate robust antigen-specific cellular immunity in mice and may potentially be useful for control of SARS-CoV infection. PMID- 15823605 TI - Vpu-mediated CD4 down-regulation and degradation is conserved among highly divergent SIV(cpz) strains. AB - Human immunodeficiency virus type 1 (HIV-1) along with simian immunodeficiency viruses from chimpanzees (SIV(cpz)) and three species of Old World monkeys from the genus Cercopithecus have been shown to encode a Vpu protein. To date, the functional characterization of Vpu has been limited to a small number of subtype B and more recently subtype C Vpu proteins. Using a recently developed VpuEGFP reporter system, we have shown that the subtype B and C Vpus are capable of preventing CD4 from being expressed on the cell surface. Using the same reporter system, we report here on the expression and functional analysis of Vpu protein from four SIV(cpz) isolates (CAM13, ANT, TAN1, and GAB1). All four SIV Vpu fusion proteins were efficiently expressed and prevented CD4 expression on the cell surface and induced CD4 degradation. This was surprising as three of the SIV(cpz) Vpu fusion proteins had only one canonical casein kinase II (CK-II) site (CAM13, ANT, TAN1) while previous studies with laboratory adapted HXB2 had indicated that both CK-II sites were required for CD4 degradation. Both ANT and TAN1 Vpu sequences encoded five consecutive negatively charged amino acids residues following the only CKII site (SAIEEDEE for ANT; SGVEEDEE for TAN1). We thus explored the possibility that this stretch of negatively charged amino acids might substitute for the lack of second CK-II site. Substitution of the aspartic acid at position 61 and glutamic acid at position 63 in the SIV(cpz) ANT Vpu within with lysine residues abolished the ability of this protein to down modulate cell surface expression of CD4. Similarly, change of a serine to an alanine residue following the single consensus CK-II site of the CAM13 Vpu (SGNESDGGEEE) abolished CD4-down-regulation, suggesting that this serine was phosphorylated in the absence of a canonical CK-II site. Our results indicate that the serine was required, suggesting that this serine was phosphorylated by CK-II or possibly another cellular kinase. Taken together, these results show for the first time that Vpu proteins from SIV(cpz) isolates, although quite diverse in sequence and predicted secondary structure from the HIV-1 subtype B protein, are capable of down-regulating CD4, which is one of the major functions of the HIV-1 protein. PMID- 15823606 TI - The baculovirus anti-apoptotic protein Op-IAP does not inhibit Drosophila caspases or apoptosis in Drosophila S2 cells and instead sensitizes S2 cells to virus-induced apoptosis. AB - The Op-IAP protein from the baculovirus Orgyia pseudotsugata M nucleopolyhedrovirus (OpMNPV) is highly effective at inhibiting apoptosis triggered by a variety of different stimuli in lepidopteran cells as well as in several different mammalian cell types, suggesting that it functions at a highly conserved step in the apoptotic pathway. However, the mechanism by which Op-IAP inhibits apoptosis is unclear. Since some IAP proteins can bind and inhibit caspases, we tested whether Op-IAP could inhibit the activity of caspases from Drosophila melanogaster. We found that recombinant Op-IAP protein was not able to bind or directly inhibit the activity of the Drosophila caspases DRONC, DrICE, or DCP-1 in vitro. In addition, expression of Op-IAP was unable to inhibit apoptosis triggered by either actinomycin D or UV light in D. melanogaster S2 cells. Surprisingly, Op-IAP expression in S2 cells enhanced apoptosis caused by baculovirus infection, but did not cause increased sensitivity to either actinomycin D or UV damage-induced apoptosis. The observation that Op-IAP cannot inhibit these insect caspases suggests that it functions by a mechanism that does not involve direct caspase inhibition. PMID- 15823607 TI - Role of La autoantigen and polypyrimidine tract-binding protein in HCV replication. AB - To determine if the cellular factors La autoantigen (La) and polypyrimidine tract binding protein (PTB) are required for hepatitis C virus (HCV) replication, we used siRNAs to silence these factors and then monitored their effect on HCV replication using quantitative RT-PCR. In addition, we determined the influence of PTB on the activity of the 3' noncoding region (NCR) of HCV and investigated its interaction with the components of the HCV replicase complex. We found that La is essential for efficient HCV replication while PTB appears to partially repress replication. PTB does, however, block the binding of HCV RNA-dependent RNA polymerase (RdRp, NS5B) to the 3'NCR. Indirect immunofluorescence microscopy showed co-localization of cytoplasmic PTB with the HCV RdRp in hepatoma cells (Huh-7) expressing HCV proteins, while in vitro translation of viral proteins from the HCV replicon revealed the interaction of PTB isoforms with NS5B polymerase and NS3. PMID- 15823608 TI - Evaluating the immunogenicity and protective efficacy of a DNA vaccine encoding Lassa virus nucleoprotein. AB - Several viruses in the Arenavirus genus of the family Arenaviridae cause severe, often fatal, hemorrhagic fever. One such virus, Lassa virus (LV), is a frequent cause of disease in Africa, and survivors often are left with substantial neurological impairment. The feasibility of protective immunization against LV infection, and the associated disease, has been demonstrated in animal models, using recombinant vaccinia viruses to deliver Lassa proteins. Circumstantial evidence implicates cellular immunity in this Lassa-induced protection, but this has not been confirmed. Here, we describe DNA vaccines that encode LV proteins. A single inoculation of a plasmid encoding full-length Lassa nucleoprotein (LNP) can induce CD8(+) T cell responses in mice and can protect against challenge with two arenaviruses, lymphocytic choriomeningitis virus (LCMV) and Pichinde virus (PV). A DNA minigene vaccine encoding a 9 amino acid sequence from LNP also induces CD8(+) T cells and protects against arenavirus challenge, thus confirming prior speculation that protective cellular immunity is induced by LV proteins. PMID- 15823609 TI - Differential expression of domain III neutralizing epitopes on the envelope proteins of West Nile virus strains. AB - Neutralization of flaviviruses by antibody is primarily mediated via epitopes in the viral envelope (E) protein. Comparative studies using neutralizing monoclonal antibodies revealed differential expression of epitopes within the E protein domain III of ten naturally occurring West Nile virus strains representing major subtypes of genetic lineages 1 and 2. Residues that defined these subtype specific determinants were identified by mutational studies and found to be surface exposed in the domain III structure. Mutations of residue 332 had the most significant effects on variation of domain III neutralizing epitopes among strains. PMID- 15823610 TI - Comparison of DNA polymerase activities between recombinant feline immunodeficiency and leukemia virus reverse transcriptases. AB - In this study, we present enzymatic differences found between recombinant RTs derived from feline leukemia virus and feline immunodeficiency virus. Firstly, FIV RT showed low steady state K(m) values for dNTPs compared to FeLV RT. Consistent with this, FIV RT synthesized DNA more efficiently than FeLV RT at low dNTP concentrations. We observed similar concentration-dependent activity differences between other lentiviral (HIV-1 and SIV) and non-lentiviral (MuLV and AMV) RTs. Second, FeLV RT showed less efficient misincorporation with biased dNTP pools and mismatch primer extension capabilities, compared to FIV RT. In M13mp2 lacZalpha forward mutation assays, FeLV RT displayed approximately 11-fold higher fidelity than FIV RT. Finally, FeLV RT was less sensitive to 3TCTP and ddATP than FIV RT. This study represents the comprehensive enzymatic characterization of RTs from a lentivirus and a non-lentivirus retrovirus from the same host species. The data presented here support enzymatic divergences seen among retroviral RTs. PMID- 15823611 TI - Tomato spotted wilt virus transcriptase in vitro displays a preference for cap donors with multiple base complementarity to the viral template. AB - Transcription of segmented negative-strand RNA viruses is initiated by cap snatching: a host mRNA is cleaved generally at 10-20 nt from its 5' capped end and the resulting capped leader used to prime viral transcription. For Tomato spotted wilt virus (TSWV), type species of the plant-infecting Tospovirus genus within the Bunyaviridae, cap donors were previously shown to require a single base complementarity to the ultimate or penultimate viral template sequence. More recently, the occurrence in vitro of "re-snatching" of viral mRNAs, i.e., the use of viral mRNAs as cap donors, has been demonstrated for TSWV. To estimate the relative occurrence of re-snatching compared to snatching of host mRNAs, the use of cap donors with either single, double, or multiple complementarity to the viral template was analyzed in pair-wise competition in TSWV in vitro transcription assays. A strong preference was observed for multiple-basepairing donors. PMID- 15823612 TI - Growth sensitivity of a recombinant simian virus 5 P/V mutant to type I interferon differs between tumor cell lines and normal primary cells. AB - A paramyxovirus SV5 mutant (rSV5-P/V-CPI-) that encodes 6 naturally-occurring P/V gene substitutions is a potent inducer of type I interferon (IFN) and is restricted for low moi growth, two phenotypes not seen with WT SV5. In this study, we have compared the IFN sensitivity of WT SV5 and the rSV5-P/V-CPI- mutant in tumor cell lines and in cultures of normal primary cells. We have tested the hypothesis that differences in IFN induction elicited by WT rSV5 and rSV5-P/V-CPI- are responsible for differences in low moi growth and spread. In contrast to WT SV5, low moi infection of A549 lung carcinoma cells with rSV5-P/V CPI- resulted in a plateau of virus production by 24-48 h pi when secreted IFN levels were between approximately 100 and 1000 U/ml. Gene microarray and RT-PCR analyses identified IFN genes and IFN-stimulated genes whose expression were increased by infection of A549 cells with WT and P/V mutant viruses. Restricted low moi growth and spread of rSV5-P/V-CPI- in A549 cells was relieved in the presence of neutralizing antibodies to IFN-beta but not TNF-alpha. When A549 or MDA-MB-435 breast tumor cells were pretreated with IFN, both WT and P/V mutant viruses showed delayed spread and approximately 10-fold reduction in virus yield, but infections were not eliminated. Using normal primary human epithelial cells that have undergone limited passage in culture, WT rSV5 and rSV5-P/V-CPI- displayed high moi growth properties that were similar to that seen in A549 cells. However, IFN pretreatment of these primary cells as well as normal human lung cells eliminated low moi spread of both mutant and WT rSV5 infections. Together, these data demonstrate that SV5 growth in normal primary human cells is highly sensitive to IFN compared to growth in some tumor cell lines, regardless of whether the P/V gene is WT or mutant. These results suggest a model in which spread of WT SV5 in normal human cells is dependent on the ability of the virus to prevent IFN synthesis. The implications of these results for the use of recombinant paramyxoviruses as vectors are discussed. PMID- 15823613 TI - Progesterone synthesis by the human placenta. AB - One of the essential roles of the human placenta is to produce the steroid hormone progesterone, which is required for the maintenance of pregnancy. The rate-determining step of placental progesterone synthesis is the conversion of cholesterol to pregnenolone by cytochrome P450scc (CYP11A1) in placental mitochondria in a reaction requiring electrons delivered via adrenodoxin reductase and adrenodoxin. Pregnenolone is converted to progesterone by type 1 3beta-hydroxysteroid dehydrogenase located in the mitochondrion. Progesterone synthesis by the human placenta displays notable differences from steroid synthesis in the classical steroid producing tissues such as the adrenal cortex and corpus luteum. One important difference is that the placenta lacks short term modulation of steroid synthesis and does not express the steroidogenic acute regulatory (StAR) protein. The most notable difference between the placenta and other steroidogenic tissues is that electron supply to P450scc limits the rate at which cholesterol is converted to pregnenolone in the placenta. The limiting component for electron delivery to P450scc is the concentration of adrenodoxin reductase in the mitochondrial matrix which is insufficient to maintain the adrenodoxin pool in a fully reduced state. Evidence suggests that placental mitochondria have a near-saturating cholesterol concentration for P450scc, likely provided by the StAR-like protein MLN64, and cholesterol translocation to the P450scc is not a major site of regulation of progesterone synthesis. Cyclic AMP stimulates progesterone synthesis by the human placenta but uncertainty remains regarding the key hormones that control cyclic AMP levels. The mechanism of regulation of adrenodoxin reductase levels in the human placenta remains to be studied. PMID- 15823614 TI - Immunocytological evidence for hematopoiesis in the early human placenta. AB - Hematopoiesis has previously been observed in the human yolk sac, in placental villi and in the embryonic aorta. Here, our immunocytological study at 24 and 35 days showed packed erythroblasts in the placental vessels, mitotic figures and anti-Ki-67 reactions within these cells. Morphologically, the erythroblasts and vessels were similar to those found in the yolk sac during primitive hematopoiesis. In addition, numerous extravascular erythroblasts were found in the villous core. Positive reactions were obtained in erythroblasts using antibodies against glycophorin-A, GATA-2 and C-kit that characterize the hematopoietic cells. However, erythroblasts did not react with anti-CD34 and anti CD45. In this respect, they differ from the hematopoietic cell clusters observed in the aorta of the human embryo. The staining for glycophorin-A was maintained in erythroblasts at 6-7 weeks and 12-14 weeks. Anti-GATA-2 reaction was decreased in erythroblasts and appeared in the perivillous cytotrophoblast. Anti-C-kit signal was detected in endothelial cells at 6-7 weeks and switched to stromal and perivascular cells at 12-14 weeks. By term, anti-GATA-2 staining was still present in the trophoblast and appeared in vessels while anti-C-kit was negative. For the leukocytes marker CD15, a staining was found in the endothelium at 35 days, 6-7 and 12-14 weeks and in leukocytes at term. CD45 antibody decorated the leukocytes at 12-14 weeks and at term. Erythroblasts undergo a primitive hematopoiesis in the early placental vessels that may be of value for the embryo in a period of low oxygen environment. PMID- 15823615 TI - Localization of prostaglandin H synthase, prostaglandin dehydrogenase, corticotropin releasing hormone and glucocorticoid receptor in rhesus monkey fetal membranes with labor and in the presence of infection. AB - Prostaglandins (PGs) play a central role in primate parturition by their actions on uterine contractility and on cervical ripening. Rhesus monkey placentation is hemochorial and the endocrine events surrounding parturition are qualitatively similar to human pregnancy. Although there is an increase in PG production before the onset of labor, little is known about the cellular localization of the PGH synthase (PGHS) or the 15-hydroxy PG dehydrogenase (PGDH) in the fetal membranes of nonhuman primates and whether it changes at term in spontaneous labor or during preterm labor associated with infection. Placental corticotropin releasing hormone (CRH) and the glucocorticoid receptor (GR) have also been implicated as mediators in parturition by virtue of their roles in PG production. We utilized immunohistochemical methods to localize the inducible isoform PGHS-2, PGDH, GR and CRH in rhesus monkey amnion, chorion and attached decidua. Tissues were obtained at cesarean section during late pregnancy, in spontaneous labor at term and in premature labor induced by Group B streptococcal intraamniotic infection. Specific staining for immunoreactive (ir)-PGHS-2 was observed in amnion epithelial and mesenchymal cells and to a lesser extent in chorion and decidua. In contrast, ir-PGDH was localized primarily to the extravillous trophoblast layer of chorion. GR was localized to both the cytoplasm and nucleus of amnion epithelial cells, subepithelial fibroblasts, chorion trophoblasts and in decidua. Immunostaining for CRH was found in amnion and in scattered decidual cells but was most intense in the chorion trophoblast layer. There was no demonstrable change in this overall pattern of immunostaining in association with the onset of labor at term except for a decrease in staining for ir-PGDH in chorion. Experimental Group B streptococcal chorioamnionitis resulted in preterm labor and extensive necrosis of extravillous trophoblast cells with subsequent loss of chorionic ir-PGDH and relative sparing of ir-PGHS-2 in amnion epithelium which favors the net production of PGs. The expression pattern of these effectors in the rhesus monkey fetal membranes points to a functional role of PGs and glucocorticoids in the process of term and preterm parturition which is similar to that in human pregnancy. PMID- 15823616 TI - Expression of variant luteinizing hormone/chorionic gonadotropin receptors and degradation of chorionic gonadotropin in human chorionic villous macrophages. AB - Human gonads and non-gonadal organs/tissues express luteinizing hormone/chorionic gonadotropin (LH/CG) receptors. This study aimed to identify the LH/CG receptors and to clarify their function in human placental chorionic villous macrophages. Macrophages as well as syncytiotrophoblasts of human chorionic villous tissues were immunohistochemically positive for LH/CG receptor throughout gestation. By reverse transcription-nested polymerase chain reaction methods, villous macrophages were shown to express a variant type of LH/CG receptor, the sequencing of which revealed a deletion of exon 9. For experiments in vitro, a monocyte-macrophage cell line, THP-1, was transfected with vector alone, wild type LH/CG receptor, and exon 9-deleted LH/CG receptor after phorbol 12-myristate 13-acetate (PMA) treatment. Non-PMA-treated THP-1 cells transfected with vector alone were also examined. THP-1 cells expressed exon 9-deleted LH/CG receptor after treatment with PMA. After the cells of the four groups were cultured in medium containing intact human CG (hCG), the concentrations of hCG and its beta core fragment (beta-CF) were measured in the supernatant of the culture medium and in the cell cytosol. Time-dependent hCG uptake was observed in both non-PMA treated and PMA-treated THP-1 cells, suggesting that the variant receptor is not directly involved in the ingestion of hCG. The degradation of hCG and excretion of beta-CF were progressed in PMA-treated cells but not in the un-treated cells. In the cell cytosol, the ratio of beta-CF and hCG concentrations (beta-CF/hCG) was significantly higher in the PMA-treated cells than in non-PMA-treated cells; however, it did not differ between the PMA-treated cells transfected with exon 9 deleted receptor and those transfected with vector alone. Macrophages may express the variant receptor in order to recognize the intracytoplasmic hCG and transport it to the lysosome. Among the two PMA-treated cells, the ratio was lower in those transfected with wild-type receptor. The expression of the variant receptor may modulate the degradation of hCG but be reduced by expression of the wild-type receptor in its lacking macrophages. Our data suggest a potentially important role for exon 9-deleted LH/CG receptors expressed in human placental villous macrophages in the local metabolism of hCG. PMID- 15823617 TI - Flow cytometric characterisation of cells of differing densities isolated from human term placentae and enrichment of villous trophoblast cells. AB - Cells were isolated from human term placentae by trypsinisation of fragments of chorionic villi and fractionation of cells on a Percoll density gradient into six layers. A panel of 10 monoclonal antibodies to antigens on or in trophoblast cells (placental alkaline phosphatase (PLAP), cytokeratin-7, beta-human chorionic gonadotrophin (beta-hCG), human leucocyte antigen-G (HLA-G)), leucocytes (CD45), monocytes, macrophages, dendritic cells, B cells (HLA class II), mesenchyme cells (vimentin), fibroblasts (fibroblast antigen) and nucleated cells excluding villous trophoblast (HLA class I, CD9) was used to characterise the cells by flow cytometry. For staining intracellular antigens (cytokeratin, vimentin, beta-hCG) the cells were first fixed and permeabilised. The upper two layers from the gradient (density 1.013-1.039 g/ml) contained predominantly PLAP-positive cells or fragments, probably derived from the syncytiotrophoblast. Cytokeratin-positive cells accumulated mainly in the layer of density 1.039-1.052 g/ml and comprised the majority of the cell types identified in this fraction. Few or no cells reactive with antibodies to beta-hCG or HLA-G were identified in any layer. Non trophoblast cells were heavier, being present mainly at densities 1.052-1.079 g/ml (CD45, HLA class I, vimentin) and 1.066-1.092 g/ml (fibroblast). Fewer than 10% of cells in any layer were HLA class II- or CD9-positive. Further purification of trophoblast cells was by negative immunomagnetic separation with removal of CD45-positive cells and HLA class II-positive cells to less than 1%. On culture of the cells from each layer, those of density 1.039-1.066 g/ml exhibited characteristics of cytotrophoblast cells; they secreted high levels of human chorionic gonadotrophin and formed adherent multinucleate cells. This procedure enabled the selection and enrichment of cytotrophoblast cells and/or syncytiotrophoblast fragments that are suitable for cellular and molecular studies. PMID- 15823618 TI - Microarray-based identification of differentially expressed genes in hypoxic term human trophoblasts and in placental villi of pregnancies with growth restricted fetuses. AB - Hypoxia adversely influences the function of the human placenta. We sought to identify a set of hypoxia-regulated transcripts in both term human trophoblasts in vitro and in villous trophoblasts in vivo. Using high-density oligonucleotide microarrays we initially examined differences in gene expression between trophoblast cultured in standard conditions (20% oxygen) vs. hypoxic conditions (< or =1% oxygen), as well as in placental tissues from pregnancies complicated by intrauterine growth restriction vs. matched controls. We used a novel computation method to compile data from the two approaches and identify transcripts that exhibited a marked expression change. Using quantitative PCR we confirmed an up-regulation of transcripts for vascular endothelial growth factor, connective tissue growth factor, follistatin-related protein, N-Myc down regulated gene 1 and adipophilin in hypoxic term trophoblasts. In contrast, the expression of human placental lactogen and Beckwith-Wiedemann region 1 C was reduced in hypoxic trophoblast. Using in situ hybridization we validated the expression of each transcript in cultured term villous trophoblasts, and determined transcript expression in placental samples derived from four sets of dichorionic twins complicated by growth restriction of one twin. The identification of hypoxic trophoblast signature transcripts may implicate new mediators in pathways underlying trophoblast hypoxic injury and adaptation. PMID- 15823619 TI - Fetal growth restriction is associated with increased apoptosis in the chorionic trophoblast cells of human fetal membranes. AB - The aim of the study was to determine the incidence and spatial distribution of apoptotic cell in fetal membranes obtained from human pregnancies complicated with fetal growth restriction (FGR) for which there was no established cause. Fetal membrane samples from normal (n=10) and FGR-affected (n=10) pregnancies were collected and stored following delivery. The incidence of apoptosis and the number of apoptotic cells in normal and FGR-affected fetal membranes were determined using immunohistochemistry and a monoclonal antibody for neo-epitope of cytokeratin-18, M30. The level of apoptotic proteins in FGR-fetal membranes compared to the normal tissue was determined using Western immunoblotting analysis. Multiple labeling of trophoblast cells using immunofluorescence markers was used to investigate regional differences in localization of apoptotic cells between normal and FGR-affected fetal membranes. Apoptosis was detected in both normal and FGR-affected fetal membranes. However, quantitative analysis of apoptotic cells by immunohistochemistry showed a significant increase in FGR affected fetal membranes compared to normal (p<0.005). Furthermore, it was observed that apoptotic cells were predominantly localized to chorio-decidual layer of the fetal membrane. By using semi-quantitative analysis of Western immunoblotting, a significant increase in the levels of the apoptotic marker proteins poly-ribo (ADP) polymerase (PARP) and the neo-epitope of cytokeratin-18 were observed in FGR-affected fetal membranes compared to normal (p<0.005). Immunofluorescence studies further confirmed the restriction of the apoptotic cells to the chorionic trophoblast cells in FGR-affected fetal membranes. Our results document for the first time an increased incidence of apoptosis in FGR affected fetal membranes, with the apoptotic cells restricted primarily to the chorionic trophoblast layer of the fetal membranes. Increased apoptosis in FGR affected fetal membranes may impair functions of the fetal membranes that are important for normal fetal development and growth. Elucidation of the molecular mechanisms involved in the control of apoptosis in the chorionic trophoblast layer in the FGR-affected fetal membranes may provide further insights into the etiology of FGR. PMID- 15823620 TI - Increased matrix metalloproteinases 2 and 9 in placenta of diabetic rats at midgestation. AB - Matrix metalloproteinases (MMPs) are involved in placental remodelling throughout pregnancy. Diabetes mellitus induces alterations in tissue production of NO, a regulator of MMPs activity. The present work evaluates placental and fetal MMPs and NO levels during midpregnancy in neonatal streptozotocin-induced diabetic rats. MMP-2 and MMP-9 immunolabelling was increased both in the labyrinth zone (p<0.001) and in the giant trophoblast cells of the junctional zone (p<0.001) from diabetic placenta, when compared with controls. Also MMP-2 (p<0.01) and MMP 9 (p<0.005) activities were increased in both maternal and fetal sides of diabetic placenta when related to controls. In both sides of the diabetic placenta, nitrate/nitrite concentrations (which indicate NO production) were higher than in controls (p<0.05). An intense immunostaining for nitrotyrosine, indicating peroxynitrite-induced damage, was found in both labyrinth (p<0.001) and junctional zones (p<0.001) of diabetic placenta. Enhanced MMP-2 activity (p<0.05) and NO production were also higher in the fetuses from diabetic rats when compared to controls (p<0.005). These findings demonstrate alterations in MMPs and NO in the feto-placental unit of diabetic rats, anomalies that are likely to be involved in the developmental alterations induced by maternal diabetes. PMID- 15823621 TI - Electrical potential difference between mother and conceptus in the mouse. AB - This study aimed to determine whether there is a maternofetal potential difference (PD) in the mouse. The mean (+/- SEM) in vivo electrical potential difference (PD(sa)) between saline filled catheters in the maternal subcutaneous space (s) and the fetal amniotic sac (a) measured, according to strict criteria, in anaesthetised MF1 mice at a gestational age of 18-20 (term 20) days was 3.9+/ 0.5 mV (significantly different from zero P<0.0001) in 16 conceptuses from 11 mice with the amniotic sac positive with respect to the maternal catheter. The PD(sv) between maternal tail vein (v) and maternal subcutaneous space was -0.8+/ 0.4 mV (n=3: not significantly different from zero). Measurement of PD between two different maternal subcutaneous catheters (n=4) was < or =0.5 mV. This study shows that there is a maternofetal PD in the mouse and provides the foundation for studies addressing its mechanism of generation in this species. PMID- 15823622 TI - Dissolved oxygen concentration in culture medium: assumptions and pitfalls. AB - Oxygen is a key factor in the regulation of cytotrophoblast differentiation, proliferation and invasion in early pregnancy. Abnormalities in oxygen concentration have also been linked to a number of pregnancy disorders. Cell culture models have been used to study the effect of oxygen on cytotrophoblast behaviour in vitro, however, there is often little or no validation of oxygen levels in these cell culture systems. In this study, dissolved oxygen levels in culture medium maintained in standard culture conditions (18% O(2)) measured 18%. On transfer to a low oxygen environment (2% O(2)), oxygen levels decreased to 6 8% after 4h and reached 2% only after 24h in culture. Culture medium pre-gassed with nitrogen to remove dissolved oxygen quickly absorbed oxygen when exposed to ambient air during dispensing and required further incubation in a 2% oxygen environment before dissolved oxygen levels equilibrated to 2%. Thus, cultured cells placed in a low oxygen environment would be exposed to varying levels of oxygen before the desired level of oxygen exposure is reached. This study highlights the importance of validation of oxygen levels and potential problems associated with in vitro studies on the regulatory effects of oxygen. PMID- 15823623 TI - Myocardial ischemia/reperfusion-injury, a clinical view on a complex pathophysiological process. AB - Myocardial infarction is the major cause of death in the world. Over the last two decades, coronary reperfusion therapy has become established for the management of acute myocardial infarction (AMI). However, restoration of blood flow to previously ischemic myocardium results in the so-called ischemia/reperfusion (IR) injury. The different clinical manifestations of this injury include myocardial necrosis, arrhythmia, myocardial stunning and endothelial- and microvascular dysfunction including the no-reflow phenomenon. The pathogenesis of ischemia/reperfusion injury consists of many mechanisms. Recently, there's increasing evidence for an important role in IR-injury on hypercontracture induced by high levels of cytosolic calcium or by low concentrations of ATP. In the last years, many studies on experimental models were investigated, but the clinical trials confirming these effects remain spare. Recently, the beneficial effect of Na(+)/H(+)-exchange inhibitor cariporide and of the oxygen-derived free radical (ODFR) scavenger vitamin E on coronary bypass surgery-induced IR-injury were demonstrated. Also recently, the beneficial effect of allopurinol on the recovery of left ventricular function after rescue balloon-dilatation was demonstrated. The beneficial effect of magnesium and trimetazidine on IR-injury remains controversial. The beneficial effect of adenosine remains to be further confirmed. There's also increasing interest in agentia combining the property of upregulating NO-synthase (e.g. L-arginine) and restoring the balance between NO and free radicals (e.g. tetrahydrobiopterin). One of such agents could be folic acid. In this review article the authors give an overview of the recent insights concerning pathogenesis and therapeutic possibilities to prevent IR-induced injury. PMID- 15823624 TI - Antithrombotic strategies for the management of non-valvular atrial fibrillation. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia, affecting an estimated 2.2 million Americans. The management of non-valvular AF is likely to become even more challenging over the coming decades. The treatment target of AF takes 3 forms: cardioversion, rate control, and minimization of embolic risk. Recently, rate, rather than rhythm control has emerged as a valid therapeutic option in patients with persistent AF. According to current guidelines, International Normalized Ratio-targeted oral anticoagulation and proper stratification of risk remain fundamental principles of management. This article reviews, in depth, the current therapeutic options for atrial fibrillation including oral anticoagulants and unfractionated heparin, as well as reviewing new therapeutic options including bridging therapy with low-molecular-weight heparins and the new oral antithrombotic drug, ximelagatran. PMID- 15823625 TI - MORE--MOexipril and REgression of left ventricle hypertrophy in combination therapy A multicentric open label clinical trial. AB - AIM: To evaluate the effect of ACE inhibitor moexipril if added to combination therapy in patient with poorly controlled hypertension. PATIENTS: Four hundred twenty patients with hypertension treated with monotherapy or two dug combination without an ACE inhibitor or AII antagonist and with blood pressure >/=140/90 mm Hg. DESIGN: Single-blind, multicenter, open, with a double-blind echocardiographic examination. METHODS: Basic cardiological examination including echocardiography was performed before including into the study. If the patient fulfilled inclusion criteria, ACE inhibitor moexipril was added to the therapy and uptitrated according to BP values. BP measuring, clinical examination, and basic laboratory were performed every month, echocardiography was repeated after 6 months. RESULTS: Sitting BP decreased from 161.43+/-12.84/96.72+/-7.74 mm Hg to 135.87+/-9.98/82.36+/-5.83 mm Hg (p<0.0001), heart rate from 73.08+/-9.87 to 69.80+/-7.91 (p<0.0001). Three hundred forty patients (81%) had BPd <90 mm Hg after 6 months. Left ventricle mass decreased from 263.24+/-94.69 to 246.71+/ 89.08 g (p<0.0001), left atrium decreased from 39.78+/-5.40 to 38.89+/-4.98 mm (p<0.0001), and E/A ratio increased from 0.91+/-0.28 to 0.94+/-0.27 (p<0.0005). Plasma cholesterol level decreased from 5.67+/-0.87 to 5.44+/-0.68 mmol/l (p<0.0001) and plasma triglycerides decreased from 1.92+/-1.07 to 1.78+/-0.80 mmol/l (p<0.001). A greater effect on blood pressure reduction was observed in combination ACE-I+diuretics than in combination ACE-I+betablocker or ACE-I+Ca blocker (statistically borderline). A statistically greater effect on left ventricle mass was observed if moexipril was added to a diuretic than to Ca blocker (p=0.02) or betablocker (p=0.04). SUMMARY: ACE inhibitor moexipril added to combination therapy of hypertension had similar effect on blood pressure reduction and left ventricle mass as in monotherapy trials. The most effective combination is ACE ihibitor+thiazide diuretic. A very small number of adverse events was observed; cough was reported in 2.14% of patients. Decreased heart rate and improvement in lipid parameters were observed in the whole group. PMID- 15823626 TI - Soluble vascular endothelial growth factor, soluble VEGF receptor Flt-1 and endothelial function in healthy smokers. AB - OBJECTIVE: To relate levels of vascular endothelial growth factor (VEGF) and its soluble receptor, sFlt-1, with endothelial function in healthy smokers. METHODS: Plasma levels of VEGF and sFlt-1 were measured by ELISA in 22 healthy smokers and 22 matched healthy non-smoking controls, and compared to flow- (FMD) and acetylcholine-mediated (AMD) vasodilatation (endothelial-dependent) (EDV) and nitroglycerine-mediated (NMD) vasodilatation (endothelial-independent) of lower extremities were measured with plethysmography. RESULTS: Smokers and controls had similar plasma VEGF levels, but sFlt-1 levels were lower in smokers than in controls (p<0.01). AMD was lower in smokers compared to controls (p<0.05), but FMD and NMD levels were similar. Smokers and controls with high AMD (>12 ml/100 ml tissue/min) had significantly lower plasma VEGF levels (p<0.001). An inverse correlation was found in both groups, between VEGF and AMD (smokers: r=-0.6, p<0.01; controls: r=-0.71, p<0.005) and with FMD (smokers: r=-0.56, p<0.05; controls: r=-0.58, p<0.005). There were no significant correlations between sFlt 1 with VEGF levels or endothelial-dependent dilatation. CONCLUSION: In conclusion, healthy smokers demonstrate abnormal AMD, and an inverse correlation between plasma VEGF levels (but not sFlt-1) with indices of endothelial dysfunction (FMD and AMD) exists. VEGF, and not sFlt-1, may be related to the pathogenesis of endothelial dysfunction in healthy smoking individuals. PMID- 15823627 TI - Ratings of positive and depressive emotion as predictors of mortality in coronary patients. AB - BACKGROUND: An extensive body of research has demonstrated an association between negative affective states and health outcomes. Positive emotions may also influence physical health, however, their examination has received far less attention. METHODS: Positive and depressive emotion ratings were examined as independent and joint predictors of survival in a sample of 866 cardiac catheterization patients. The mean follow-up was 11.4 years, during which 415 deaths occurred. The mean age of the sample was 60.3 years and 74.3% of the sample were males. RESULTS: Both positive and depressive ratings were associated with survival (hazard ratio=0.80, 95% CI=0.66-0.97, p<0.025 and hazard ratio=1.32, 95% CI=1.13-1.54, p<0.001, respectively), adjusted for risk factors. Positive emotion ratings were no longer significantly related to mortality when the two emotion measures were simultaneously included in a survival model. CONCLUSION: The current findings are consistent with a theory that suggests that the relation between positive emotion and mortality may be partly mediated by depressive emotion. PMID- 15823628 TI - Effects of weight loss and risk factor treatment in subjects with elevated serum C3, an inflammatory predictor of myocardial infarction. AB - BACKGROUND: Serum C3 is an inflammatory predictor of myocardial infarction and a covariate of fasting insulin and several endogenous risk factors. This study was performed to ascertain whether risk factor control may reduce elevated C3 concentrations. METHODS: After traditional risk factor and C3 assessment in 1100 unselected men aged 55-64 years, 238 men with persistently elevated C3 levels (>=1.19 g/l, high tertile) were randomised into 2 groups: 43 controls, who were referred to their general practitioner, and 195 subjects who were intensively treated with diet, and anti-hypertensive or antidiabetic drugs according to specific indications, without anti-dyslipidemic drugs. RESULTS: After three months in the treated subjects significant decrements of body weight, blood pressure, blood glucose and serum lipids were obtained, with stable C3 levels (while in controls a 3.3% increase occurred, P=0.02). The factors associated with a C3 decrement >5% were a high baseline C3 level, a recent acute inflammation, physical activity, belonging to the treated group, and a significant reduction in body weight, triglycerides or blood glucose. However, in multivariate analysis only an elevated baseline C3 (P<0.0001), a weight loss >2% (P=0.0009) and physical activity (P=0.02) remained independently associated with a C3 decrement >5% (R(2)=0.14). CONCLUSIONS: Only weight loss and physical activity, but not traditional risk factor lowering, could independently induce a significant C3 decrease. Thus, C3 elevation is associated with, but probably not caused by, traditional risk factors. PMID- 15823629 TI - Evolution of vascular brachytherapy over time: data from the RENO-registry analysis. AB - OBJECTIVE: To study the evolution of procedural variations in vascular brachytherapy (VBT) and their relationship to medium-term outcome. METHODS AND RESULTS: The RENO (European Surveillance Registry with Novoste Beta-Cath) prospectively collected procedural and clinical outcome data on 1098 patients treated with VBT. Patients were divided for this analysis into Group-I, the first 50% registered, and Group-II, the last 50% registered. Shorter 30-mm source trains were more commonly used in Group-I (p<0.001) while longer 40-mm (p=NS) and 60-mm (p<0.001) source trains were more commonly used in Group-II. Mean dwell time for radiation seeds was longer in Group-II compared to Group-I (4.20+/-1.48 min vs. 4.14+/-1.44 min; p<0.05). Mean radiation dose was higher in Group-II (19.73+/-3.33 Gy vs. 17.92+/-2.68 Gy; p<0.001). Cutting balloons were more frequently used in Group-II (p<0.001). There was significant drop in the incidence of geographic miss in Group-II (3.2% vs. 9%; p<0.00005). There were nonsignificant trends towards reduction in angiographic restenosis, target vessel (TV) revascularisation, death and major adverse cardiac events (MACE). CONCLUSION: There has been a learning curve and evolution of VBT techniques over time. In general, there has been an increase in radiation source length, use of cutting balloons, dwell time and radiation dose. This has resulted in significant reduction of geographic miss and a trend towards improve clinical outcomes. Continued development may result in further improvement in the treatment of patients with in-stent restenosis (ISR). PMID- 15823630 TI - Management of non-ST-elevation acute coronary syndromes: how cost-effective are glycoprotein IIb/IIIA antagonists in the UK National Health Service? AB - BACKGROUND: The glycoprotein IIb/IIIa antagonists (GPAs) represent a new class of drugs to prevent platelet aggregation in the acute treatment of non-ST-elevation acute coronary syndromes (NSTE-ACS). Systematic reviews have identified serious limitations in published cost-effectiveness analyses, including a lack of UK specific studies and an absence of studies comparing different protocols for the use of GPAs. METHODS: A model was developed to assess the cost effectiveness of a variety of protocols employing GPAs for patients presenting with NSTE-ACS in the UK. The perspective of the UK National Health Service was adopted, with outcomes in terms of quality-adjusted life-years (QALYs). Four treatment strategies were evaluated: GPAs as part of initial medical management (Strategy 1); GPAs in patients with planned percutaneous coronary interventions (PCIs; Strategy 2); GPAs as an adjunct to the PCI procedure (Strategy 3); and no GPAs (Strategy 4). Baseline event rates and costs were taken from a UK observational study of ACS patients and relative risk reductions from GPAs were taken from a meta analysis of trials. Long-term costs and QALYs were estimated using data from a UK longitudinal study. RESULTS: The most cost-effective use of GPAs is likely to be Strategy 1, with an incremental cost per QALY gained of between pound4605 to pound10,343. Focusing this use of GPAs only on the subgroup of patients at high risk appears to represent the most cost-effective use of NHS resources. CONCLUSIONS: Medical management of patients with NSTE-ACS using GPAs is the most cost-effective use of resources, particularly if targeted to higher risk subgroups. PMID- 15823631 TI - Persistent smokers after myocardial infarction: a group that requires special attention. AB - BACKGROUND: Despite the detrimental effects of smoking on the cardiovascular system, a significant number of patients with coronary heart disease continue to smoke. We aimed to record compliance to medication and attitude towards recommended lifestyle changes in patients who suffered from myocardial infarction (MI) and continued to smoke after the coronary event. METHODS: A total of 1011 consecutive patients (<75 years) with a history of MI (>6 months) were recruited during the period 2000-2003 from the outpatient cardiology clinic of a district general hospital. All patients were interviewed and blood was taken for lipid measurements. Glycosylated haemoglobin (HbA(1 c)) was also measured in all diabetics. RESULTS: Three hundred and twenty-nine (32.5%) patients reported smoking at interview, while 338 (33.5%) were ex-smokers of whom 278 (45.8% of all smokers) had quit smoking after MI and 344 (34%) had never smoked. Persistent smokers had significantly lower high-density lipoprotein cholesterol levels than nonsmokers (1.03+/-0.28 vs. 1.09+/-0.29 mmol/l, p=0.001). Persistent smokers with diabetes had poorer glycaemic control than nonsmoker diabetic patients as indicated by HbA(1c) levels (8+/-1.7% vs. 7.2+/-1.3%, p=0.001). Fewer persistent smokers were taking hypolipidaemic drugs than nonsmokers (31% vs. 40.3%, p=0.005). Finally, persistent smokers were less frequently performing regular exercise than nonsmokers (42% vs. 51%, p=0.008). CONCLUSIONS: Patients who remain smokers after MI have a more negative attitude towards health aspects, are less compliant with their medications, and therefore constitute a high-risk subgroup, which requires special attention and should be professionally encouraged and supported to stop smoking. PMID- 15823632 TI - Autonomic modulation and clinical outcome in patients with chronic heart failure. AB - OBJECTIVE: Several studies documented the relevance of autonomic activity in the pathophysiology of heart failure. In our study we evaluated the adjustment of this activity under different stimuli, by means of heart rate variability (HRV), and correlated these findings with long-term mortality and sustained VT occurrence. PATIENTS AND METHOD: Fifty-three patients (mean age 54+/-9 years) with heart failure were submitted to time and frequency domain HRV analysis. This latter analysis was performed at rest, during paced breathing and during passive tilt. RESULTS: Lower standard deviation of RR intervals (76.76+/-24 versus 107.70+/-43, p=0.02), mean of the 5-min standard deviations of RR intervals (35.14+/-15 versus 62.39, p<0.01), standard deviation of the 5-min average RR intervals (69.42+/-19 versus 91.79+/-30, p=0.02), and baseline Low-Frequency (LF) power (15.15+/-12 versus 40.39+/-24 nu, p=0.001) characterized patients who died. Paced breathing induced a significant reduction of LF (40.39+/-24 to 20.12+/-18 nu, p<0.0001) and increase of High Frequency power (HF) (47.31+/-23 to 70.63+/-16 nu, p<0.0001) in survivors, while tilting induced a reduction of HF (47.31+/-23 to 29.80+/-16 nu, p<0.0001). Patients who died did not show significant variation of HRV neither during paced breathing nor during tilt. Reduced time domain indexes were significantly correlated to sustained VT occurrence. CONCLUSIONS: Patients with heart failure with a better prognosis are characterized by a responsiveness of autonomic modulation. Simple maneuvers, such as tilting and paced breathing, seem to provide more useful information, than the baseline evaluation of autonomic status, in identifying patients with a higher mortality. Time domain analysis was more helpful to estimate arrhythmic risk. PMID- 15823633 TI - Epoetin alfa's effect on left ventricular hypertrophy and subsequent mortality. AB - BACKGROUND: Improving anemia in patients with chronic renal failure (CRF) and congestive heart failure (CHF) also improves left ventricular hypertrophy (LVH). No previous meta-analyses have been conducted to further examine this association, including the association between LVH and mortality in these patients. METHODS AND RESULTS: Literature searches on MEDLINE, EMBASE, and OVID were performed using Cochrane Library protocols. Two hundred sixteen abstracts were reviewed preliminarily for inclusion in the meta-analysis of epoetin alfa, anemia and 5 pre-selected parameters of LVH. One hundred seventy-nine abstracts were reviewed for LVH and mortality. The predominant hematologic and left ventricular function changes observed during epoetin alfa treatment in patients with CHF and CRF are (1) increases in hemoglobin (Hb) and hematocrit (Hct); (2) decreases in left ventricular mass (LVM) and LVM index; (3) increase in ejection fraction (EF); and (4) decreases in left ventricular end-diastolic and end systolic volume. Three independent factors-target Hb, duration of disease, and duration of follow-up-each had a statistically significant association with Hb, Hct, and EF, respectively. A separate meta-analysis using 3 risk models showed LVH is strongly and positively associated with both cardiovascular and all-cause mortality, with two- to three-fold increases in risk. CONCLUSIONS: LVH is common in patients with CRF and CHF. Current findings indicate epoetin alfa therapy results in anemia amelioration, as evidenced by higher Hb and Hct levels, and reduction of key LVH parameters. LVM regression is associated with lower incidence of cardiovascular-related morbidity and mortality, therefore epoetin alfa therapy may provide a survival benefit. PMID- 15823634 TI - Safety and efficacy of Sildenafil therapy in children with pulmonary hypertension. AB - OBJECTIVE: Sildenafil is a selective Phosphodiesterase-5 inhibitor that has been reported to be a potent pulmonary vasodilator. We evaluated the safety, efficacy and pharmacokinetics of oral Sildenafil in a case series of children with pulmonary hypertension. METHODS: Three children, 1 with primary pulmonary hypertension (patient 1) and 2 with pulmonary hypertension associated with congenital heart disease (patients 2 and 3) were enrolled. Sildenafil was started at 0.5 mg/kg 4-hourly and the dose increased to 1.0 and then to 2.0 mg/kg/dose. Patients were assessed at baseline and then monthly for a total of 6 visits. RESULTS: All patients reported increased exercise capacity with improvement in New York Heart Association functional class. The distance walked during the 6-min test increased by 74% (patient 1), 75% (patient 2) and 25% (patient 3) and oxyhaemoglobin saturations increased from 79%, 97% and 80% to 93%, 100% and 93%, respectively. There were no side effects and no fall in systemic blood pressure. Sildenafil plasma levels 1 h after a 0.5, 1.0 and 2 mg/kg dose of Sildenafil were 109+/-87, 150+/-62 and 368+/-200 ng/ml, respectively. They fell to 211+/-106 ng/ml 3 h after the 2.0 mg/kg dose. CONCLUSIONS: Medium term Sildenafil therapy improves oxyhaemoglobin saturations and exercise tolerance in children with pulmonary hypertension without any side effects. Mean plasma levels 1 h after doses of 0.5-2.0 mg/kg are similar to the maximum plasma concentrations reported in adults receiving doses within the therapeutic range. Sildenafil use in children appears to be safe and may be beneficial in the management of pulmonary arterial hypertension. PMID- 15823635 TI - Early determination of homocysteine levels in acute coronary syndromes, is it an independent prognostic factor? AB - INTRODUCTION: Increased concentrations of homocysteine (tHcy) are considered a potentially modifiable risk factor for coronary heart disease. The relationship between plasma homocysteine and prognosis has been less well studied. The aim of this study was to examine a possible relationship between the homocysteine levels in admission and all cause mortality in subjects presenting with non-ST segment elevation (NSTE) acute coronary syndrome. METHODS: We studied 854 patients with suspected NSTE acute coronary syndrome admitted consecutively to our institution, tHcy was determined at a median of 3 days from enrolment and was analyzed in tertiles together with classical risk factors and other biochemical markers. The primary end point was all cause mortality at 1 year follow-up. RESULTS: There were 86 deaths in the upper 2 tertiles (> or =10.1 mmol/L). The events registered in the lower tertile of admission homocysteine concentration were 12 deaths. Therefore, tHcy values over 10 mumol/l increases the posibility of long term all cause mortality after an NSTE acute coronary syndromes (HR 2.5). This is independent of other prognostic factors as important as age, cardiovascular risk factors, congestive heart failure or creatinine levels at arrival. This is the first study that shows the tHcy prognostic value with independece of the acute phase reactants. CONCLUSION: Determination of plasmatic levels of tHcy in the acute phase of a NSTE acute coronary syndrome is a useful tool in the prognostic stratification, independently of classical risk markers (age, cardiovascular risk factors, heart failure, troponin peak) of acute phase reactants and of creatinine obtained at arrival. PMID- 15823636 TI - Prevalence and characteristics of diabetic patients in a chronic heart failure population. AB - BACKGROUND: Previously, estimates of the prevalence of diabetes mellitus (DM) in patients with chronic heart failure (CHF) have solely been based on history in retrospective studies. The aim of this study was to investigate the prevalence of DM and glucose abnormalities in patients with CHF in accordance with the modern diagnostic criteria of DM. A second aim was to characterize patients with DM with respect to severity of CHF. METHODS AND RESULTS: A prospective study including patients with systolic CHF admitted to a heart failure clinic. Fasting blood glucose (FBG) levels were measured on two occasions, for classification of DM. Severity of CHF was assessed by NYHA class, echocardiography, 6 min walk test and plasma NT-proBNP levels. A total of 195 consecutive patients were included in the study, of these 188 were eligible for classification of DM. A total of 21% had DM by history, 11% of the patients without known DM had FBG> or =6.1 mmol/l, and a diagnosis of DM was confirmed in half of these patients. There were no differences in severity of heart failure between patients with DM by definition and non-DM patients; however, high body mass index was related to worse CHF symptoms. CONCLUSION: Twenty six percent of the patients had DM by definition, 20% of these were undiagnosed at the time of admission. One single measurement of FBG is not sufficient to establish a diagnosis of DM. Diabetes was not related to CHF symptoms or to a degree of left ventricular dysfunction. PMID- 15823637 TI - Hemoglobin levels and 30-day mortality in patients after myocardial infarction. AB - BACKGROUND: Anemia is an independent risk factor for cardiovascular (CV) outcomes in patients with coronary artery disease and heart failure. However, the effect of hemoglobin levels on short-term CV mortality in patients with acute myocardial infarction (MI) remains unclear. METHODS: In a retrospective study we analyzed 1841 consecutive patients admitted with the diagnosis of acute MI. The primary end-point of the study was 30-day mortality. Patients were categorized according to the hemoglobin level on admission (10 g/dl or less, or greater than 10 g/dl). RESULTS: The overall 30-day mortality was 10.3%. The mortality was 21.6% in patients with hemoglobin levels on admission < or =10 g/dl and 9.3% in patients with hemoglobin levels >10 g/dl (p<0.001). Multivariate logistic regression analysis showed, that lower hemoglobin concentration is an independent predictor of 30-day mortality, when adjusted for other risk factors (HR 1.76, CI 1.08-2.85; p=0.02). CONCLUSIONS: Lower levels of hemoglobin are associated with higher short term mortality in patients with acute MI. Specific therapeutic strategies in anemic patients with MI should be further considered. PMID- 15823638 TI - The effect of physical training on workload, upper leg muscle function and muscle areas in patients with chronic heart failure. AB - OBJECTIVE: To investigate the effect of physical training (PTr) on upper leg muscle area, muscle strength and muscle endurance expressed as upper leg muscle function (ULMF) in relation to exercise performance in CHF. DESIGN: Randomised to a training (TG) or control group (CG). SETTING: Outpatient cardiac rehabilitation centre of community hospital. PATIENTS: 77 CHF patients (59 men and 18 women), NYHA class II/III, age 59.8+/-9.3 years, LVEF 27+/-8%. Sixteen patients dropped out during the intervention period, 61 patients (M/F:46/15) completed the study. INTERVENTION: PTr (combined strength and endurance exercises) four times per week, twice supervised and twice at home, during 26 weeks. MAIN OUTCOME MEASURES: LVEF, body composition, daily physical activity, exercise performance, upper leg muscle area and isokinetic leg muscle variables. RESULTS: Workload and peak oxygen consumption decreased in the CG (-4.1% and -4%) but increased in the TG (+5% and +4%) following PTr (p<0.05, ANOVA repeated measures). Hamstrings area decreased in the CG and did not change in the TG (p<0.05, ANOVA repeated measures). ULMF improved in the TG, but remained unchanged in the CG (+13.0% and 0.0, respectively, p<0.05; ANOVA repeated measures). At baseline and after intervention nearly 60% of the variance in maximal workload was explained by ULMF and quadriceps muscle area (multiple regression analysis). CONCLUSIONS: In CHF patients, home-based training in conjunction with a supervised strength and endurance training program is safe, feasible and effective and does not require complex training equipment. Physical training prevented loss of hamstrings muscle mass and improved exercise performance by enhancing muscle strength and endurance. PMID- 15823639 TI - Real-time monitoring of coronary care mortality: a comparison and combination of two monitoring tools. AB - BACKGROUND: Quality control in clinical care is becoming increasingly more prevalent, resulting in a need for tools that can be readily used by clinical teams for monitoring their own performance. The aim of this study was to devise a practical and effective scheme for monitoring coronary care mortality in real time. METHODS: We obtained data for 2153 consecutive patients admitted after acute myocardial infarction between 1st September and 30th November 1995 to one of 20 acute hospitals in West Yorkshire participating in the NHS R and D funded EMMACE-1 study. We developed control charts for each centre to monitor 30-day mortality. These control charts used two complementary tools: the Risk-Adjusted Cumulative Sum chart (CUSUM) and a new 'Rocket Tail' chart based upon the Variable Life-Adjusted Display (VLAD). We also combined information from each of these to devise a further chart. RESULTS: Control charts are shown for two centres in order to illustrate the important features of the different but complimentary monitoring tools. The Risk-Adjusted CUSUM is shown to be useful for detecting isolated runs of unsatisfactory outcome results but is not 'intuitive', and does not give any indication of the preceding history of outcomes. The Rocket Tail chart is shown to give a good summary of outcome history and also how overall performance compares with what would be expected for the case-mix. A chart that combines both approaches appeals to the advantages of each. CONCLUSIONS: We propose a visual approach to health-care monitoring that beneficially combines and extends the different information of the previously used CUSUM and VLAD charts. PMID- 15823640 TI - Effects of switching statins on lipid and apolipoprotein ratios in the MERCURY I study. AB - BACKGROUND: Lipid ratios are clinically useful markers of coronary artery disease (CAD) risk. The effects of rosuvastatin, atorvastatin, simvastatin, and pravastatin on lipid ratios were investigated in the Measuring Effective Reductions in Cholesterol Using Rosuvastatin TherapY (MERCURY) I trial. METHODS: This trial was conducted in 3140 hypercholesterolemic patients with CAD, atherosclerosis, type 2 diabetes mellitus, or a 20% 10-year risk for CAD. Patients were randomized to rosuvastatin 10 mg, atorvastatin 10 or 20 mg, simvastatin 20 mg, or pravastatin 40 mg for 8 weeks; all patients except those receiving rosuvastatin 10 mg either were switched to rosuvastatin 10 or 20 mg or remained on initial treatment for 8 more weeks. RESULTS: At 8 weeks, reductions in total cholesterol (TC):high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol:HDL-C, non-HDL-C:HDL-C, and apolipoprotein (apo) B:apo A-I ratios with rosuvastatin 10 mg were significantly greater than those with atorvastatin 10 mg, atorvastatin 20 mg, simvastatin 20 mg, and pravastatin 40 mg (P<0.0001 for all). At week 16, switching to rosuvastatin 10 mg from atorvastatin 10 mg, simvastatin 20 mg, and pravastatin 40 mg and to rosuvastatin 20 mg from atorvastatin 20 mg produced significantly greater reductions in all lipid ratios (P< or =0.0001 for all). Switching to rosuvastatin 10 mg from atorvastatin 20 mg produced significantly greater reductions in TC:HDL-C (P<0.025) and apo B:apo A-I (P<0.01). CONCLUSIONS: Rosuvastatin 10 mg reduces lipid ratios more than equivalent and higher doses of other statins; switching to equal or lower doses of rosuvastatin produces significantly improved reductions in lipid ratios. PMID- 15823641 TI - P wave signal averaged ECG and chemoreflexsensitivity in paroxysmal atrial fibrillation. AB - Detailed analysis of the QRS-complex and autonomic dysfunction can identify patients at risk to suffer from ventricular arrhythmias. To determine whether patients at risk for paroxysmal atrial fibrillation (PAF) could be identified while in sinus rhythm, a P wave triggered signal averaged ECG and an analysis of the autonomic function by chemoreflexsensitivity (CHRS) were examined. The ratio between the difference of RR intervals in the ECG and the venous partial pressure of oxygen before and after 5-min oxygen inhalation was measured for the determination of CHRS. We examined 224 patients (group A) who suffered from PAF, 250 patients (group B) without arrhythmic history and 30 young volunteers (group C). The filtered P wave duration (FPD) was significantly longer in group A than in group B (140.9+/-21.0 vs. 118.2+/-9.4 ms, p<0.0001) or C (105.2+/-14.1 ms, p<0.0001) while the root mean square voltage of the last 20 ms of the P wave (RMS 20) was significantly lower in group A than in group B (2.68+/-1.12 vs. 4.06+/ 1.57 microV, p<0.0001) or C (3.97+/-1.36 microV, p<0.0001). Atrial late potentials (ALP) were defined as a FPD>120 ms and a RMS 20< or =3.5 microV. ALP could identify patients of group A with a specificity of 78% and a sensitivity of 83%. Patients with PAF (2.32+/-1.15 ms/mm Hg) showed a significantly lower CHRS than group B (4.14+/-1.58 ms/mm Hg, p<0.0001) or group C (4.98+/-1.51 ms/mm Hg, p<0.0001). The sensitivity for the presence of atrial fibrillation was 71% for a CHRS below 3.0 ms/mm Hg with a specificity of 70%. A combination of both methods showed a specificity of 85% and a sensitivity of 65% when ALP and pathological CHRS were present. The results of our study suggest that risk of atrial fibrillation could be detected by P wave signal averaged ECG and CHRS. An analysis of CHRS seems to be an appropriate method to demonstrate a neurovegetative imbalance, which might be one possible trigger mechanism. PMID- 15823642 TI - The clinical relevance of raised cardiac troponin I in the absence of significant angiographic coronary artery disease. AB - OBJECTIVES: Cardiac troponins are highly sensitive and specific markers of myocardial cell injury. We wished to determine the clinical relevance of raised troponins in the absence of significant angiographic coronary artery disease. DESIGN AND METHODS: We assessed patients admitted to our hospital over the past 3 years with troponin-positive chest pain and no angiographically significant coronary disease. RESULTS: The study included 67 patients, all of whom had symptoms of "chest pain" and elevated (>0.2 microg/L) troponin I on admission. Thirty-four (51%) patients had alternative causes for myocyte injury other than coronary ischaemia. In the remaining 33 (49%) patients we could find no other associated features or diagnoses. Follow up was obtained in 29 (88%) of these 33 patients (mean follow up 58+/-13 weeks, range 17-156 weeks). During the follow up period, three (4.5%) patients were readmitted with further ischaemic events. CONCLUSIONS: Myocardial damage can occur in the absence of significant angiographic coronary disease and other causes of raised troponins should be considered according to the clinical presentation. Troponin-positive cases with angiographically "normal" coronary arteries can re-present with future cardiac events and should still be considered for aggressive risk management. PMID- 15823643 TI - Definite trial is not definitive after all. PMID- 15823644 TI - One-child policy and increased mechanization are additional risk factors for increased coronary artery disease in modern China. PMID- 15823645 TI - Limited effect of early repair on carotid arterial wall stiffness in adult post coarctectomy patients: in response to the article by Heger M, Willfort A, Neunteufl T, Rosenhek R, Gabriel H, Wollenek G, Wimmer M, Maurer G, Baumgartner H. Vascular dysfunction after coarctation repair is related to the age at surgery. Int J Cardiol 2004; in press. PMID- 15823646 TI - Vagal dysfunction and the pathogenesis of chronic Chagas disease. AB - Vagal dysfunction is thought to be an early, primary and specific abnormality of chronic Chagas disease. However, chagasic patients with unequivocal evidence of heart disease, can have normal or abnormal vagal control of heart rate. A common explanation for these apparently discordant and contradictory results is proposed. PMID- 15823647 TI - Late recurrence of outflow tract obstruction seven years after septal ablation in hypertrophic cardiomyopathy. AB - We present a case of late recurrence of outflow tract obstruction 7 years after successful septal ablation in hypertrophic cardiomyopathy. In addition the debate over maximal versus minimal treatment is discussed. PMID- 15823648 TI - Returning hypertrophy after surgery in a patient with hypertrophic cardiomyopathy caused by a myosin-binding protein C mutation. AB - We report a 13-year follow-up of a patient who underwent both myectomy and septal ablation due to hypertrophic cardiomyopathy caused by a cardiac myosin-binding protein C gene mutation. After myectomy the patient again developed significant septal hypertrophy at the operated septal area with a need for a second interventional therapy. This exceptional case underscores the remarkable ability of the heart muscle to show a continuous hypertrophic process over many years. PMID- 15823649 TI - Bacterial resistance to silver in wound care. AB - Ionic silver exhibits antimicrobial activity against a broad range of micro organisms. As a consequence, silver is included in many commercially available healthcare products. The use of silver is increasing rapidly in the field of wound care, and a wide variety of silver-containing dressings are now commonplace (e.g. Hydrofiber dressing, polyurethane foams and gauzes). However, concerns associated with the overuse of silver and the consequent emergence of bacterial resistance are being raised. The current understanding of the biochemical and molecular basis behind silver resistance has been documented since 1998. Despite the sporadic evidence of bacterial resistance to silver, there have been very few studies undertaken and documented to ascertain its prevalence. The risks of antibacterial resistance developing from the use of biocides may well have been overstated. It is proposed that hygiene should be emphasized and targeted towards those applications that have demonstrable benefits in wound care. It is the purpose of this review to assess the likelihood of widespread resistance to silver and the potential for silver to induce cross-resistance to antibiotics, in light of its increasing usage within the healthcare setting. PMID- 15823650 TI - Epidemiological investigation of a Serratia liquefaciens outbreak in a neurosurgery department. AB - Between February 2001 and March 2003, 17 patients from the neurosurgery department of the University Hospital of Rangueil (Toulouse, Southern France) developed Serratia liquefaciens infections. Due to the atypical antibiotype displayed by the clinical isolates (i.e. gentamicin resistance), an outbreak was suspected. Molecular analysis carried out by pulsed-field gel electrophoresis demonstrated a genetic link for all patients. Furthermore, the patient who introduced the epidemic Serratia strain was also identified and shown to be related to the two epidemic peaks observed during the outbreak period. Investigation failed to reveal a reservoir among the antiseptics and soaps, or among the mechanical ventilators used. However, when the colonization of patients was investigated, positive carriage was observed and could be considered as a potential risk for the spread of the epidemic strain. Due to the delay between antibiotherapy and S. liquefaciens colonization, a selection effect had to be considered. Finally, implementation of hygiene measures was accompanied by control of the outbreak. PMID- 15823651 TI - Investigation of a nosocomial outbreak of extended-spectrum beta-lactamase VEB-1 producing isolates of Acinetobacter baumannii in a hospital setting. AB - A nosocomial outbreak of epidemiologically related VEB-1 extended-spectrum beta lactamase-producing isolates of Acinetobacter baumannii occurred in 33 patients in an intensive care unit. A case-control study identified previous treatment with third-generation cephalosporins as the only risk factor for A. baumannii acquisition. Rationale for antibiotic use should be strengthened. PMID- 15823652 TI - Identification of an imipenem-resistant Pseudomonas aeruginosa clone among patients in a hospital in Rio de Janeiro. AB - A total of 85 Pseudomonas aeruginosa isolates were obtained from October 1999 to April 2000 in a tertiary care hospital in Rio de Janeiro, Brazil. The imipenem susceptibility was evaluated by disk diffusion and agar dilution methods, and the clonal relationship among 67 isolates was examined by macrorestriction profile analysis following pulsed-field gel electrophoresis. Imipenem resistance was observed in 52 (61.2%) isolates. Imipenem-resistant P. aeruginosa isolates were separated into 10 genotypes, 73% of which belonged to genotype A. Identification of a single P. aeruginosa clone with a high rate of imipenem resistance emphasizes the need to control the transmission of this organism among patients. PMID- 15823653 TI - Estimation of methicillin-resistant Staphylococcus aureus transmission by considering colonization pressure at the time of hospital admission. AB - Our objective was to evaluate the accuracy of a methicillin-resistant Staphylococcus aureus (MRSA) rate using the imported MRSA reservoir identified at the time of hospital admission. Two indicators were used: the number of imported MRSA patient-days/total number of patient-days [representing colonization pressure (CP) at the time of admission] and the incidence of hospital-acquired MRSA isolated from clinical samples expressed as density/100 patient-days for carriers identified at the time of admission [representing the incidence taking CP into account (ICP)]. The variations of these indicators were analysed and compared with two more common indicators: percentage of MRSA acquired in our hospital and the incidence of hospital-acquired MRSA isolated from clinical samples expressed as density/1000 patient-days within three four-month periods during 2002. Common indicators varied similarly, with marked decline during the third period; first-period CP was twice that of other periods (P<10(-6)) and the highest (>two-fold) ICP was seen in the summer (second) period (P<0.001) when the personnel/patient ratio was the lowest. Thus, comparison of different indicators within four-month periods underlines important differences between common and novel indicators. Despite several limitations, ICP should be helpful in the interpretation of MRSA surveillance data, particularly for estimating the extent of MRSA transmission. PMID- 15823654 TI - Microbiological evaluation of two hand hygiene procedures achieved by healthcare workers during routine patient care: a randomized study. AB - The aim of this study was to evaluate the comparative microbiological efficacy of hand rubbing and handwashing in healthcare workers from different wards, with particular emphasis on transient flora, and to assess predisposing factors for hand contamination after patient care in everyday practice. Over a six-month period, 50 healthcare workers were randomly assigned, using a crossover design, to perform handwashing with unmedicated soap and hand rubbing with an alcoholic solution following a healthcare procedure. Imprints of palms and fingertips were taken separately before and after each hand hygiene procedure. The number of colonies per plate was counted and transient pathogens were identified. Risk factors for hand contamination were determined. Hand rubbing produced a significantly greater reduction in microbiological load than handwashing (P<0.0001 for palms and P=0.0003 for fingertips). In multivariate analysis, working in a medical ward rather than in an intensive care unit was significantly associated with increased hand contamination (P=0.03 for palms and P=0.02 for fingertips). Transient pathogens were found on 15% of healthcare workers' hands before hand hygiene. The only factor associated with hand contamination by transient pathogens was the absence of gloving during the healthcare procedure (odds ratio 4.8; 95% confidence interval 1.2-19; P=0.03). After hand rubbing, no transient pathogens were recovered, while these were found in two cases after handwashing. Hand rubbing is more efficacious than handwashing for the decontamination of healthcare workers' hands following contact with patients and patients' environments. Gloving may reduce microbiological hand contamination by transient pathogens. PMID- 15823655 TI - Prevalence of nosocomial infections in hospitals in Norway, 2002 and 2003. AB - The Norwegian Institute of Public Health initiated a national surveillance system for nosocomial infections in 2002. The system is based on two annual one-day prevalence surveys recording the four most common types of nosocomial infection: urinary tract infections; lower respiratory tract infections; surgical site infections and septicaemia. All acute care hospitals in Norway (N=76) were invited to participate in the four surveys in 2002 and 2003. The total prevalence of the four recorded nosocomial infections varied between 5.1% and 5.4% in the four surveys. In all surveys, nosocomial infections were located most frequently in the urinary tract (34%), followed by the lower respiratory tract (29%), surgical sites (28%) and septicaemia (8%). The prevalence surveys give a brief overview of the burden and distribution of nosocomial infections. The results can be used to prioritize further infection control measures and more detailed incidence surveillance of nosocomial infections. PMID- 15823656 TI - Prospective surveillance for healthcare-associated infections in German nursing home residents. AB - We undertook a prospective surveillance study in order to determine the incidence of healthcare-associated infections (HCAIs) in German nursing home residents. All people residing for more than one day in a 103-bed nursing home for the elderly in Bonn, Germany between December 1998 and November 1999 were included. Active surveillance was based on previously published consensus definitions. Rates for HCAIs and urinary tract infections (UTIs) were calculated based on resident-days and device-utilization days, respectively. The overall incidence of HCAIs was 6.0 per 1000 resident-days, with respiratory tract infections, gastroenteritis, skin/soft tissue infections and UTIs representing 94% of all HCAIs (2.2, 1.2, 1.2 and 1.0 infections per 1000 resident-days, respectively). Residents with pneumonia were more likely to die than residents with other HCAIs (RR=5.09; 95%CI 1.87-13.89; P=0.011). We conclude that HCAIs are a serious health problem in German nursing home residents. Standardized surveillance in nursing homes is important to assess the effectiveness of infection control standards, and should be based on consensus definitions in order to allow for meaningful interfacility comparisons. In Germany, the implementation of a hospital reimbursement system based on diagnosis-related groups is likely further to increase the proportion of vulnerable populations in long-term-care facilities. PMID- 15823657 TI - Ralstonia pickettii and Burkholderia cepacia complex bloodstream infections related to infusion of contaminated water for injection. AB - Ralstonia pickettii and Burkholderia cepacia complex isolates are causes of healthcare-associated infection related to contamination of intravenously administered products. Based on microbiological and epidemiological data and molecular typing by pulsed-field gel electrophoresis, we report the occurrence of two outbreaks of R. pickettii and B. cepacia complex bloodstream infections. The first outbreak occurred from August 1995 to September 1996, and the second outbreak occurred from 28 March to 8 April 1998, affecting adults and neonates, respectively. Infusion of contaminated water for injection was the source of infection. PMID- 15823658 TI - Subtyping of Clostridium difficile polymerase chain reaction (PCR) ribotype 001 by repetitive extragenic palindromic PCR genomic fingerprinting. AB - Fifty isolates of the most common UK strain of Clostridium difficile [polymerase chain reaction (PCR) ribotype 001] were analysed by three PCR-based typing methods in order to determine genomic diversity within this strain that may form the basis of a subtyping method. The three methods used were repetitive extragenic palindromic elements (REP), conserved repetitive DNA elements (BOX), and enterobacterial repetitive PCR intergenic consensus sequences (ERIC). The performance of each typing method was assessed by comparing powers of discrimination, typeability and reproducibility. All methods had satisfactory levels of typeability and reproducibility as determined by blind-coded repeats, but REP-PCR typing proved to be the most discriminatory method, yielding seven distinct amplicon profiles consisting of up to eight major bands. BOX-PCR generated between two and five major amplicons with four distinct BOX profiles. ERIC-PCR primers, however, could not discriminate between isolates. These results suggest that PCR ribotype 001 is not clonal in nature at present, and that REP PCR subtyping methods offer promise to further our understanding of the epidemiology of C. difficile PCR ribotype 001 disease in UK hospitals. PMID- 15823659 TI - Epidemiological study of invasive pulmonary aspergillosis in a haematology unit by molecular typing of environmental and patient isolates of Aspergillus fumigatus. AB - In order to determine the possible relationship between environmental contamination by Aspergillus fumigatus and occurrence of invasive aspergillosis, a one-year prospective study was carried out in the haematology ward of Hautepierre Hospital, Strasbourg, France. During the study period, 21 environmental isolates and 26 clinical isolates of A. fumigatus were collected. Each was genotyped using a random amplification of polymorphic DNA (RAPD) technique. Thirty-four distinct profiles were identified by RAPD analysis, indicating the great genetic diversity of A. fumigatus isolated from infected patients and from the environment. For two patients, RAPD analysis demonstrated concurrent infection by at least two different strains. In two cases, a genetic similarity was noted between isolates obtained from a patient and from the environment. PMID- 15823660 TI - Water disinfection with ozone, copper and silver ions, and temperature increase to control Legionella: seven years of experience in a university teaching hospital. AB - The efficacy of ozonation, copper-silver ionization and increased temperature in controlling Legionella spp. in the hot water distribution networks of a university hospital was evaluated. Two separate water distribution networks were studied; network 1 which supplies the surgical intensive care units, and network 2 which supplies the medical intensive care units and the emergency room. Network 1 has been disinfected by ozonation since 1995, and network 2 has been disinfected by ionisation since 1999. The hot water temperature was increased from 50 to 65 degrees C in 1998 and 2000 in networks 1 and 2, respectively. Water samples and swabs of the water outlets were cultured for Legionella spp. between four and six times each year, providing data before and after implementation of the disinfection procedures. There was no significant difference in the proportion of samples positive for Legionella spp. after ozonation in network 1 or after ionization in network 2. In both networks, there was a significant reduction in legionella isolates after increasing the hot water temperature to 65 degrees C. Maintaining the hot water temperature above 50 degrees C throughout both networks proved to be the most effective control measure in our hospital. PMID- 15823661 TI - Anti-HBs profiles in children treated for neoplastic disease who had been vaccinated against hepatitis B postnatally or as infants. AB - Children with cancer are a risk group for hepatitis B virus (HBV) infection. In Poland, the initiation of a national HBV vaccination programme in neonates and infants in 1995 has contributed to the prevention of HBV infection in children treated for neoplastic diseases. The objective of this study was to analyse the anti-HBs pattern and occurrence of HBV infections in children with cancer who had been vaccinated during infancy. The study included 96 children divided into three groups: Group A, children who had received a full vaccination course with an initial level of anti-HBs >/=100 IU/L; Group B, children who had received a full vaccination course with anti-HBs <100 IU/L, for whom an additional dose of vaccine was administered; and Group C, children who had further immunoprophylaxis because they did not complete the vaccination course before cancer diagnosis. A protective level of anti-HBs after the full vaccination programme was found in 80.5% of children after three months, 74.2% after six months, 61.5% after 12 months and 78.6% after 18 months. Among children who produced antibodies, a slow decrease in the level of anti-HBs was observed. It was still protective during follow-up in Group A, but occasionally fell below 100 IU/L in Group B. In Group C, five of 11 subjects had a protective level of anti-HBs throughout follow-up. Of 28 children who showed the presence of HBsAg during follow-up, 23 eradicated the virus. In children with cancer vaccinated against HBV according to the vaccination schedule, the immune response maintains a protective level of anti HBs in more than 60% of cases, despite immunosuppression. PMID- 15823662 TI - 'Spiking' as a rapid method for differentiation of Candida albicans from other yeast species. AB - This paper describes a simple and rapid method for the differentiation of Candida albicans from other yeast species in primary cultures based on colonial morphology following incubation in carbon dioxide. The technique has superior sensitivity to the traditional germ-tube method and requires no additional laboratory tests. In a busy laboratory, this can result in significant savings in cost and time, as well as improvements in patient care. PMID- 15823663 TI - Fungal contamination of air conditioning units in operating theatres in India. AB - Postoperative fungal infections may be caused by discharge of spores from contaminated air conditioning (AC) units. The filters of such units may act as a nidus for the growth of fungi. In this survey, filters of AC units in 25 operating theatres in India were evaluated quarterly over two years. The overall rate of fungal colonization of the AC filters was 26%. In addition, window mounted AC units had higher fungal counts than wall-mounted AC units. We conclude that although the exact load of fungal spores necessary to initiate infections is unknown, AC units in operating theatres should be meticulously maintained and frequently monitored to minimize the chances of growth and proliferation of potentially pathogenic fungi. Data from this study also suggest that the design of the AC unit may play an important role in reducing fungal spore counts in the air of operating theatres in developing countries. PMID- 15823664 TI - Hydrogen peroxide fogging in an overcrowded tertiary care referral centre: some practical queries. PMID- 15823665 TI - Reducing the time interval for screening for methicillin-resistant Staphylococcus aureus. PMID- 15823666 TI - Bacteraemia with tube-coagulase-negative methicillin-resistant Staphylococcus aureus. PMID- 15823667 TI - Oral antibiotic treatment of MRSA infections. PMID- 15823668 TI - Evaluation of a hand hygiene programme on an intensive care unit. PMID- 15823669 TI - Surveillance and training, not postexposure prophylaxis, are the basis for the prevention of occupational infection by blood-borne pathogens in developing countries. PMID- 15823670 TI - CSF biomarkers for mild cognitive impairment and early Alzheimer's disease. AB - A correct clinical diagnosis, early in the course of Alzheimer's disease (AD), is of importance given the currently available symptomatic treatment with acetylcholine esterase inhibitors. The development of disease-modifying drugs like beta-sheet breakers or gamma- and beta-secretase inhibitors, emphasizes the need of improved diagnostic accuracy, especially in patients with mild cognitive impairment (MCI) that have incipient AD. Therefore, diagnostic markers in the cerebrospinal fluid (CSF) have become a rapidly growing research field. Three cerebrospinal fluid biomarkers (the 42 amino acid form of beta-amyloid (A beta), total tau, and phospho tau) have been evaluated in numerous scientific papers. These CSF markers have high sensitivity to differentiate early and incipient AD from normal aging, depression, alcohol dementia and Parkinson's disease, but lower specificity against other dementias, such as frontotemporal and Lewy body dementia. If these biomarkers are used in combination with a careful medical history, clinical examination, standard laboratory tests and imaging techniques of the brain, the diagnostic accuracy may be appropriate for the clinical evaluation of MCI cases. PMID- 15823671 TI - Diffusion tensor imaging for preoperative evaluation of tumor grade in gliomas. AB - The relationship between water diffusion parameters measured using diffusion tensor imaging (DTI) and histological malignancy of gliomas was investigated. DTI was performed using a 3.0T MR scanner in 41 consecutive patients with histologically proven gliomas. Fractional anisotropy (FA) and mean diffusivity (MD) were calculated and compared with the WHO classification of the gliomas. The FA values of grade 1 gliomas (0.150 +/- 0.017) were significantly lower than those of grade 3 (0.23 +/- 0.033) or grade 4 gliomas (0.229 +/- 0.033) (P < 0.0001, respectively). The FA values of grade 2 gliomas (0.159 +/- 0.018) were significantly lower than those of grade 3 or grade 4 gliomas (P = 0.0002, P < 0.0001, respectively). The FA threshold between low grade and high grade gliomas was 0.188. The MD values of grade 1 gliomas (1619.1 +/- 157.4 x 10(-6) mm2/s) were significantly higher than those of grade 3 (1084.5 +/- 218.9 x 10(-6) mm2/s) (P = 0.0036) or grade 4 gliomas (1098.0 +/- 291.6 x 10(-6) mm2/s) (P = 0.0002). The MD values were not correlated with the other grades of glioma. FA values can distinguish between high grade and low grade gliomas. This is useful in deciding the surgical strategy or selecting the site of stereotactic biopsy. PMID- 15823672 TI - Mitochondrial neuropathy. AB - Polyneuropathy is a frequent feature of mitochondriopathy (MCP). If and how often polyneuropathy in MCP is primarily due to the underlying disorder (mitochondrial neuropathy, MN) or due to other well-known causes is unknown. Retrospectively investigated were 108 MCP-patients with polyneuropathy. According to established diagnostic criteria 37 patients were classified as definite MCP, 56 as probable MCP and 15 as possible MCP. In 38 of the 108 MCP-patients with polyneuropathy (35%), no plausible cause for polyneuropathy other than MCP could be found. MN was characterized by weakness, muscle cramps, wasting, reduced tendon reflexes, muscle pain, ataxia, restless legs, hypesthesia, paresthesia, dysesthesia, and vegetative impairment. In 21 cases predominantly motor fibers, in 14 cases both motor and sensory fibers and in 3 cases predominantly sensory fibers were affected. Axonal degeneration was found in 19 cases, demyelination in 4 and mixed type polyneuropathy in 15. On sural nerve biopsy axonal loss was the predominant finding. In a single case tomaculae and abnormally shaped and structured mitochondria were found. MN exists, occurs in one third of the MCP-patients with polyneuropathy, and is characterized by predominant affection of the motor and sensory fibers with diffuse, symmetric and equal distribution between upper and lower limbs and by axonal degeneration. PMID- 15823673 TI - Benefits of FK506 (tacrolimus) for residual, cyclosporin- and prednisone resistant myasthenia gravis: one-year follow-up of an open-label study. AB - Thirteen patients with myasthenia gravis, unresponsive to prednisone and cyclosporin after thymectomy, received KF506 (tacrolimus) for 12 months, at starting doses of 0.1 mg/kg per day b.i.d. and then adjusted to achieve plasma concentrations between 7 and 8 ng/mL. The doses of prednisone were progressively reduced and finally discontinued. Anti-acetylcholine antibodies and myasthenia gravis score for disease severity decreased significantly and muscular strength increased by 37%. All patients achieved pharmacological remission, 11 were asymptomatic and two had minimal weakness of eyelid closure. Tacrolimus was well tolerated and appears a suitable approach after unsuccessful treatment with conventional immunosuppressants in patients with disabling myasthenia. PMID- 15823674 TI - Morphometric analysis of human occipital condyle. AB - OBJECTIVE: The human occipital condyle is the unique bony structure connecting the cranium and the vertebral column. The progress in neuroimaging techniques has increased interest for aggressive craniovertebral surgery. Such surgery requires the knowledge regarding anatomical aspects of the craniovertebral junction. The aim of the present study is to analyze the occipital condyle morphometrically. MATERIAL AND METHODS: 404 occipital condyles of 202 dry skulls were used for this study. Twenty-seven parameters were measured, including length, width and height of occipital condyle, the distances between the occipital condyle and hypoglossal canal, as well as some important condyle-related angles. RESULTS: The length, width and the height of the occipital condyle were found to be 23.4, 10.6, and 9.2 mm, respectively. The anterior and posterior intercondylar distances are 21.0 and 41.6 mm, respectively. Sagittal intercondylar angle was 59.3 degrees. The intracranial orifice of the hypoglossal canal was found in the junction of the second and third quarter on the condyle in more than 55% of specimens. The shape of occipital condyles was classified into eight types as follows--type 1: oval like condyle; type 2: kidney-like condyle; type 3: S-like condyle; type 4: eight like condyle; type 5: triangle condyle; type 6: ring-like condyle; type 7: two portioned condyle and type 8: deformed condyle. The most common type was type 1 (50%), whereas the most unusual type was type 7 (0.8%). CONCLUSION: It is concluded that the occipital condyle may present various shapes, length, width, and orientation, requiring a careful radiological analysis before craniovertebral junction surgery. PMID- 15823675 TI - Predictive value of computed tomography in visual outcome in indirect traumatic optic neuropathy complicated with periorbital facial bone fracture. AB - The visual outcome of 39 patients who had unilateral indirect type traumatic optic neuropathy and periorbital facial bone fracture treated at the Chang Gung Memorial Hospital in Kaohsiung between June 1996 and June 2001 was retrospectively evaluated, based on the results of computed tomography studies that grouped the associated fractures as anterior orbital fracture, posterior orbital fracture, orbital blow-in or blow-out fracture. Pretreatment and posttreatment visual acuities were obtained from the medical records and measurements of visual improvement degree and percentage were calculated after converting the visual acuities into log of the minimum angle of resolution (log MAR) units. This study reveals that posterior orbital fracture is associated with a worse prognosis in visual outcome than anterior orbital fracture, with visual improvement degree and percentage being 0.86 +/- 0.17 log MAR and 34.2 +/- 1.7% versus 1.56 +/- 0.27 log MAR and 58.9 +/- 1.7%, respectively (P = 0.02 and 0.004). Blow-in fractures such as an impacted lateral orbital wall did not predict significant deterioration in visual outcome. In addition, orbital floor or medial wall blow-out fractures did not predict significant improvement in visual outcome. PMID- 15823676 TI - Open-label simultaneous radio-chemotherapy of glioblastoma multiforme with topotecan in adults. AB - BACKGROUND: Due to its radioresistance, the prognosis of glioblastoma multiforme (GBM) remains poor. Therefore, we investigated the impact of simultaneous radio chemotherapy with topotecan (Hycamtin) on clinical outcome, tolerability and quality of life. PATIENTS AND METHODS: In this multicenter trial, 60 patients (19 females, 41 males) with histologically proven (5x biopsy, 31x subtotal resection, 24x total resection) GBM were included. Radio-Chemotherapy was performed with daily doses of 2.0 Gy (total, 60 Gy), and 0.5 mg (absolute dose) of topotecan intravenously 1 h prior to irradiation. Toxicity was assessed using common toxicity criteria (CTC). General condition and quality of life were assessed at baseline, at the end of therapy, and 6 weeks post-therapy. Local control and length of survival were compared with an historical control group of 67 patients only treated with postoperative radiotherapy following stereotactic biopsy (15x), subtotal resection (39x), or total resection (13x). RESULTS: 57 patients completed the therapy. Median radiation dose was 60 Gy (range 16-76 Gy). Median cumulative topotecan dose was 15 mg (range 7.5-18.5 mg). CTC toxicity grade 3 was observed in six patients and grade 4 toxicity in two patients (three events). Two patients died of septic disease. Mean Karnofsky index was 87% at baseline, 81% at the end of therapy, and 80% at 6 weeks post-therapy. Median survival time was 15 months, significantly longer than the 11 months seen in the control group (P < 0.002). Extent of tumour resection or patient age did not have a significant effect on survival. CONCLUSION: This multimodal approach is well tolerated, and quality of life remains preserved. The relatively long median survival time is promising but a further randomised double blind placebo controlled parallel designed clinical trial should be performed to confirm these results. PMID- 15823677 TI - Diagnostic and therapeutic value due to suspected diagnosis, long-term complications, and indication for sural nerve biopsy. AB - In order to elicit the usefulness of sural nerve biopsy we retrospectively evaluated the courses of disease of every patient, who underwent this procedure in our department between January 1995 and March 2000. Sixty seven patients with the suspected diagnosis of peripheral neuropathy could be included. From these chart reviews and patient questionings were done. Inflammatory-demyelinating neuropathies were suspected in 14 patients (20.9%), specific histological findings confirmed diagnosis in 50% of these patients and resulted in therapy. In cases of polyneuropathy of unknown etiology (46 patients, 68.6%) diagnosis was made in 11 patients (23.9%), and lead to therapy in 9 patients (19.6%), merely. In all, diagnostic consequences arouse in 32.8%, therapeutic consequences in 26.9%. The follow-up of 47 patients (mean 24.4 months) found chronic pain in the distribution of the sural nerve in 14 patients (29.8%), dysesthesia in 22 patients (46.8%), and persistent sensory loss in 34 patients (72.3%). Only 24 patients (51.1%) would submit to biopsy again. Because of high complication rates and poor results we conclude that sural nerve biopsy should be done only in carefully selected cases after thorough clinical work-up, and should be limited to cases of suspected inflammatory neuropathies, collagenoses and immunologic neuropathies, and hereditary neuropathies. PMID- 15823678 TI - Paced visual serial addition test in multiple sclerosis. AB - The paced auditory serial addition test (PASAT), a subtest of the multiple sclerosis functional composite score (MSFC), is increasingly used in the evaluation of cognitive function in multiple sclerosis (MS). While patient acceptance for the PASAT is low, its visual version, the paced visual serial addition test (PVSAT), is perceived to be better tolerated. The aim of this study was to investigate the interchangeability of PVSAT and PASAT in the evaluation of cognitive function in MS. Twenty-one normal controls and 50 patients with clinically definite MS were tested with PASAT and PVSAT. Both for PASAT and PVSAT, 3 and 2-second versions of two parallel test forms were used. In the PVSAT, the PASAT stimuli were shown on a computer screen. Patients were also tested with the other two MSFC subtests, i.e. the nine-hole pegboard test and timed 25-foot walk test, to calculate MSFC scores. PASAT-3 correlated highly with both PVSAT-3 and PVSAT-2. MSFC-v scores calculated with PVSAT-2 and PVSAT-3 values correlated highly with MSFC scores calculated with PASAT-3 results. The results suggest that the PVSAT can be used as an alternative for the PASAT in the MSFC. PMID- 15823679 TI - Chronic subdural haematoma: surgical treatment and outcome in 1000 cases. AB - Chronic subdural haematoma (CSDH) is one of the most common clinical entities in daily neurosurgical practice. To evaluate the clinical features, computed tomography findings, surgical results, and complications our series was statistically analysed to elucidate the factors affecting the post-operative outcome. A retrospective study (1980-2002) of the records of 1000 patients harbouring 1097 chronic subdural haematoma treated with burr-hole craniotomy with closed-system drainage was carried out. The series included 628 males and 372 females, age range 12-100 years, mean age 72.7+/-11.4 years. The mean interval from trauma to appearance of clinical symptoms was 49.1+/-7.4 days (15-751). The principal symptom was headache (29.7%) in the over 70s, and behavioural disturbance (33.8%) in the under 70s. The CSDH was right sided in 432 patients, left sided in 471, and bilateral in the remaining 97 cases. Post-operative complications occurred in 196 patients and 21 patients died in hospital. Poor prognosis was related to patient's age (>70) and clinical grade on admission (grades 0-2 versus grades 3-4). PMID- 15823680 TI - Multilevel myelopathy in Maroteaux-Lamy syndrome and review of the literature. AB - An 18-year-old male with Maroteaux-Lamy syndrome was presented with spastic quadriparesis. Magnetic resonance imaging of whole spinal canal revealed stenosis at multiple levels of cervical, thoracic and lumbar regions. By the guidance of combined evaluations of neurological examination, neuroradiological and electrophysiological findings, the most responsible spinal segment was detected each time he developed myelopathy and he underwent craniocervical, cervical and thoracolumbar decompressions consecutively. Ligamentum flavum hypertrophy was found to be the principal pathology responsible for the cord compression and myelopathy for all levels. The etiology of myelopathy and priority of the level for which decompression should be done in diffuse spinal stenosis were discussed with the literature review of Maroteaux-Lamy syndrome. PMID- 15823681 TI - Navigated dorsolateral suboccipital transcondylar (NADOSTA) approach for treatment of hypoglossal schwannoma. Case report and review of the literature. AB - Hypoglossal schwannoma is a rare skull base neoplasm, which lies in the midst of extremely complex anatomical structures. We report a 39-year-old man who presented with a history of right glossal hemiatrophy, dysesthesia and weakness of the left extremities. These symptoms were caused by a unilateral hypoglossal schwannoma located at the cranial base. This lesion was exposed through a dorsolateral suboccipital transcondylar approach, which was navigated with the frameless stereotaxy (NADOSTA). We also describe the epidemiology, symptomatology and the diagnosis of hypoglossal schwannoma as well as literature review of the operative approach. By introducing this minimally invasive dorsal surgical approach with neuronavigation in treating hypoglossal schwannoma, we believe that safe exposure and complete tumor resection can be achieved with minimal rate of morbidity. PMID- 15823682 TI - Recurrent Candida tropicalis meningitis. AB - Candida meningitis, a previously rare occurrence, has been increasing in prevalence and often is a result of complications of neurosurgery. We describe the case of a 49-year-old man who presented with headache, vertigo, intermittent blurred vision, and multiple episodes of nausea and vomiting. Computed tomography (CT) showed a left cerebellar hemorrhage with obliteration of the fourth ventricle causing hydrocephalus. He had an occipital craniotomy with transcondylar evacuation of the hemorrhage and placement of a temporary ventriculostomy. The hospital stay was prolonged because of postsurgical complications, and Candida tropicalis meningitis developed. Treatment was started with 400 mg of fluconazole administered intravenously every 12 h. In vitro susceptibility testing showed a minimum inhibitory concentration (MIC) to fluconazole of 1 microg/mL. Fluconazole was therefore continued orally for a total of 60 days, and the patient remained asymptomatic for 2 years. He then presented with increased vertigo and ataxia. Cerebrospinal fluid cultures grew C. tropicalis, which again showed susceptibility to fluconazole with a MIC of 1 microg/mL, identical to that in the previous infection. However, a second course of fluconazole failed to control the infection despite adequate cerebrospinal fluid levels. PMID- 15823683 TI - Brain stem lesion mimicking myasthenia gravis. AB - Brain stem masses may mimic Myasthenia gravis (MG), clinically and electrophysiologically. We present a 38 year-old female patient with a brain stem mass, who has clinical features similar with MG, and revealed a decremental response to repetitive nerve stimulation. Brain stem mass was removed by surgery. In the postoperative period, her complaints were regressed. Control repetitive nerve stimulation examination was normal. PMID- 15823684 TI - Idiopathic hypertrophic cranial pachymeningitis with perifocal brain edema. AB - Idiopathic hypertrophic cranial pachymeningitis (IHCP) consists of meningeal thickening due to chronic inflammation for which no cause such as infection, specific granulomatous disease, or malignancy is demonstrable. We present a case of IHCP with perifocal brain edema suggesting mass lesion with its magnetic resonance imaging (MRI) and pathologic findings. A 36-year-old woman was admitted to our hospital in August 2001 with a complaint of motor weakness in her left leg for 1 month. Magnetic resonance imaging with gadolinium revealed enhancement of the thickened dura mater and perifocal edema in the right frontoparietal region suggesting a mass lesion. Histological examination of the biopsy specimen revealed a dense fibrous cellular tissue. There was no obvious inflammatory infiltrate but in some areas between fibrous bundles one or two lymphocytes were detected. According to our knowledge, our patient is the second report in the literature of IHCP with focal edema causing a mass effect. PMID- 15823685 TI - Cervical dumbbell meningioma and thoracic dumbbell schwannoma in a patient with neurofibromatosis. AB - The occurrence of both dumbbell meningioma and dumbbell schwannoma in one patient has not been reported in the literature. We present a 16-year-old female patient, who had progressive bilateral hearing impairment for 5 years and a progressively enlarged, non-tender neck mass for 1.5 years. Mild motor weakness over her right upper limb was noted 1 week before admission. No cafe-au-lait spot was noted. Magnetic resonance imaging (MRI) revealed bilateral cerebellopontine angle tumors, a C1-2 dumbbell tumor, and a T5-6 dumbbell tumor. Neurofibromatosis type 2 was diagnosed. The cervical spine and thoracic spine tumors were removed via one-staged combined posterior (laminectomy) and antero-lateral (transforaminal or thoracoscopic) approaches during two operations performed 1 month apart. The pathology revealed meningioma and schwannoma, respectively. The patient had good recovery after these two operations and her motor function improved. Six months after the second surgery, radiosurgery was performed for the bilateral acoustic tumors, because of enlarged tumor size on follow-up MRI. To the best of our knowledge, this is the first case reported in the literature of a patient, having both dumbbell meningioma and dumbbell schwannoma. A literature review of the dumbbell tumors was done, and their treatment strategies were discussed. PMID- 15823686 TI - Cerebral Whipple's disease as a cause of reversible dementia. AB - We describe a case of reversible dementia caused by Tropheryma whippelii (TW). Diagnosis was confirmed by a positive polymerase chain reaction for this pathogen both on serum and cerebrospinal fluid. Specific antibiotic therapy resulted in distinct clinical and neuroradiological improvement. Control polymerase chain reaction for T. whippelii on serum and cerebrospinal fluid was negative. PMID- 15823687 TI - The smoking gun: many conditions associated with tobacco exposure may be attributable to paradoxical compensatory autonomic responses to nicotine. AB - Tobacco exposure is implicated in many illnesses such as cardiovascular disease and cancer, but the mechanisms underlying these associations are poorly understood. The mechanisms by which tobacco induces pro-sympathetic and pro inflammatory changes also remain elusive. Some studies have attributed these changes to the direct effects of nicotine, but such findings run counter to the pro-vagal, anti-inflammatory nature of the nicotinic pathway. We hypothesize that the illnesses associated with smoking may be partly attributable to autonomic dysfunction, sympathetic bias, and T helper (Th)2 inflammation induced by a paradoxical compensatory response to intermittent nicotinic exposure. The confusion of interpreting the adrenergia and inflammation associated with nicotine as a primary response instead of a secondary compensation may be explained by the unusually rapid absorption, action, and serum elimination of nicotine. Given the fast action and clearance of nicotine, even heavy smokers spend large portions of the day and the entire night in nicotine withdrawal, at which time rebound sympathetic bias may manifest as a result of desensitization of nicotinic receptors. This may help reconcile why the features observed in smokers such as tachycardia, hypertension, inflammation, insomnia, and anxiety, which are perhaps mistakenly attributed to the direct action of nicotine, are identical to those seen during acute nicotine withdrawal after smoking cessation. On the other hand, delayed responses to cessation of smoking such as weight gain and increased heart rate variability are compatible with reduced sympathovagal ratio and resensitization of nicotinic receptors. Sympathetic bias and the associated Th2 inflammation underlie many systemic diseases. Tobacco-related cancers may be partly attributable to immunomodulatory properties of chronic nicotine exposure by dampening Th1 immunity and enabling tumoral evasion of immune surveillance. Other conditions associated with tobacco exposure may also operate through similar autonomic and immune dysfunctions. Therapeutic implications are discussed. PMID- 15823688 TI - "Starve a fever and feed a cold": feeding and anorexia may be adaptive behavioral modulators of autonomic and T helper balance. AB - Anorexia is a common symptom accompanying infections, but the teleology of the phenomenon remains unexplained. We hypothesize that anorexia may represent a prehistoric behavioral adaptation to fight infection by maintaining T helper (Th)2 bias, which is particularly vital in fighting bacterial pathogens. Specifically, we propose that anorexia may avert the reduction of Th2/Th1 ratio by preventing feeding-induced neurohormonal and vagal output from the gut. Emerging evidence suggests that the vagal and neurohormonal output of the gut during feeding promotes Th1 function, which is desirable in fighting viral infections. Since fever may be an adaptation to fight bacteria and "colds" are generally viral in origin, the adage "starve a fever and feed a cold" may reflect a sensible behavioral strategy to tilt autonomic and Th balance in directions that are optimal for fighting the particular type of infection. The ability to modulate T helper balance through the neurohormonal and autonomic axis by adjusting food intake may be the mechanism behind other unexplained clinical observations such as the improved outcomes of ICU patients after enteric versus parenteric feedings. Compared to the prehistoric period when bacterial infection was commonplace, the anorexic response may be less adaptive today when viruses and cancers have become common triggers of anorexia. By promoting host anorexia, cachexia, and insomnia, cancers and viruses can deter behaviors such as digestion and sleep that would raise vagal and Th1 activity against tumors and viruses. Hydration and sleep, unexplained but widely accepted recommendations for flu patients, may also work by promoting vagal and Th1 functions. Modulating feeding, hydration, and sleep may prove beneficial in treating other conditions associated with abnormal autonomic and Th balance. PMID- 15823689 TI - Transplanted human embryonic stem cells as biological 'catalysts' for tissue repair and regeneration. AB - Human embryonic stem cells have tremendous potential in the newly emerging field of regenerative medicine. Recently, it was demonstrated that the rescue of lethal cardiac defects in Id knockout mutant mouse embryos was not due to the transplanted cells giving rise to functional new tissues within the defective embryonic heart. Instead, there is indirect evidence that the observed therapeutic effect was due to various secreted factors emanating from the transplanted cells. This therefore, introduces the exciting prospect of utilizing human embryonic stem cells as biological 'catalysts' to promote tissue repair and regeneration in transplantation therapy. However, the immunological barrier against allogenic transplantation, as well as the teratogenic potential of human embryonic stem cells poses major technical challenges. A possible strategy to overcome the immunological barrier may be to impose a temporary regimen of immunosuppressive drugs followed by their gradual withdrawal, once adequate tissue regeneration has been achieved. Other more novel alternatives include the use of microencapsulation to block interaction with the transplant recipient's immune system, and co-transplantation with bone marrow-derived mesenchymal stem cells, which have been demonstrated to possess immuno-suppressive properties. The teratogenic potential of human embryonic stem cells could possibly be alleviated by directing the differentiation of these cells to specific lineages prior to transplantation, or through mitotic inactivation (gamma irradiation or mitomycin C exposure). Co-transplantation with autologous adult stem cells may represent a novel strategy to further enhance the 'catalytic' effects of human embryonic stem cells. The various factors secreted by human embryonic stem cells could then have a concentrated localized effect on relatively large numbers of co-transplanted autologous adult stem cells, which may in turn lead to enhanced repair and regeneration of the damaged tissue or organ. Moreover, there is also a possibility that synergistic interactions between the co-transplanted human embryonic stem cells and autologous adult stem cells, may somehow produce signals for the recruitment and migration of additional endogenous adult stem cells within the recipient (i.e. peripheral blood circulation, bone marrow), which could further enhance organ/tissue regeneration. Hence, the potential use of human embryonic stem cells as biological 'catalysts' to stimulate tissue repair and regeneration, appears to hold tremendous promise in the field of regenerative medicine. This new therapeutic strategy needs to thoroughly investigated, in view of its potentially important clinical applications. PMID- 15823690 TI - Preventive and curative effects of probiotics in atopic patients. AB - Normally, the transport of allergens through the intestinal epithelia to the blood is limited. It is hypothesised that if these compounds arrive in the blood circulation, they must percolate through the epithelial cell layer. Thus, food allergy (and thus atopic eczema) implies an increased intercellular leakage of the gut wall. Such increased intercellular leakage is thought to be caused by a slightly changed cellular morphology due to a slight cytopathologic effect because of both a limited decay of the cytoskeleton and a slightly reduced turgor. These events may be due to a reduced production of intracellular metabolic energy in the epithelial cells due to an increased concentration of familiar, frequently occurring, potentially toxic bacterial metabolites, i.e., d lactic acid and/or ethanol. In this hypothesis we suggest that adequate probiotics can (i) prevent the increased characteristic intestinal permeability of children with atopic eczema and food allergy, (ii) can thus prevent the uptake of allergens, and (iii) finally can prevent the expression of the atopic constitution. The use of adequate probiotic lactobacilli, i.e., homolactic and/or facultatively heterolactic l-lactic acid-producing lactobacilli, reduces the intestinal amounts of the bacterial, toxic metabolites, d-lactic acid and ethanol by fermentative production of merely the non-toxic l-lactic acid from glucose. Thus, it is thought that beneficial probiotic micro-organisms promote gut barrier function and both undo and prevent unfavourable intestinal micro-ecological alterations in allergic individuals. PMID- 15823691 TI - Pathophysiology of capillary haemangioma growth after birth. AB - Capillary haemangiomas (CHs) are the most common soft tissue tumours of infancy. It is generally believed that the primary defect in CHs is intrinsic to endothelial cells, but their pathogenesis is yet poorly understood. The relatively low oxygen environment, in which the human foeto-placental unit develops, during the first trimester, is necessary to induce vasculo-angiogenesis via embryonic endothelial cells proliferation, since these cells are sensitive to hypoxia and acidosis. In newborn infants with haemangioma, persistent embryonic primitive endothelial cells trapped in the intimae underneath the developing vessels, and representing "leader" endothelial cells, can stabilise the labile vascular endothelial growth factor mRNA (VEGF mRNA), produce other angiogenic factors, degrade the underlying basement membrane and invade into the stroma of the neighbouring tissue. With bearing down, the transition from intra- to extra uterine life is accompanied by more or less pronounced hypoxia. Consequently, in babies with haemangioma, hypoxia can act as a switch to activate these "leader" endothelial cells and thereby initiate a cascade of reactions leading to CH proliferation. As they are regulated by embryonic cells, the haemangioma growth mechanisms pursue the pathway of embryonic angiogenesis and it will stop at the end of the embryonic endothelial cell cycle. Addressing this mechanism in vivo has partly been done (the angiogenic peptide bFGF varies with haemangioma growth). Thus, early treatment seems necessary in infants with haemangioma, before the endothelial cells achieve their proliferative stage. The use of an antibody to interfere with VEGF receptors provides a particular attractive strategy. PMID- 15823692 TI - Site specific therapy: an integrative approach to treating melanoma. AB - There have been many proposed theories for effectively treating melanoma, especially through the regulation of histamine. Histamine has been proven to be a major regulator of the immune system's T-helper cell subset balance and major shifts in this balance towards TH2 cytokines have contributed to diseases such as asthma, lupus and cancer. Histamine also causes suppression of interferon-induced proteins needed for anti-tumor response and activates T-suppressor cell function in cancers such as squamous cell carcinoma and melanoma. Scientific evidence has suggested the possibility of an anthistamine approach as treatment to these diseases and for melanoma, there has been great promise. This is due to the fact that melanotic cells have been elucidated to express histamine receptors and as a result, regulation of histamine could occur specifically at the site of these epidermal growths. Another factor to consider is how effective an inflammatory response can be when combined with regulation of histamine. Inflammation is a very powerful tool against pathogenic environments by causing cytokine recruitment and migration of dendritic cells to infected sites. Adequate stimulation of an inflammatory response at the specific site of any cancerous region would greatly weaken its evasive mechanisms. However, there are no reports showing high efficacy utilizing the benefits of regulating inflammation and histamine that could cause TH1 subset levels to predominate, down-regulate T suppressor cells, up-regulate interferon-induced proteins and properly sustain migration of dendritic cells concurrently. These benefits have been proven in separate instances for a range of diseases but have not been assessed as a combined modality for melanoma therapy. Therefore successful melanoma treatment should integrate these principles involving: the use of H2 antagonists for preventing the negative effects of histamine, monoclonal antibodies to ensure an effective dendritic cell response, and routine pro-inflammatory induction at the specific site of the melanotic tissue to ensure recognition of the cancer that has evaded immunity. PMID- 15823693 TI - The possible use of arsine (AsH3) as therapy for malaria. AB - Arsine has long been recognized as a selective toxin that targets the membrane of erythrocytes and causes leakage of the internal contents of these cells. In acute poisoning episodes, the victims usually sustain kidney malfunction resulting from collection of debris from the erythrocytes. The hypothesis presented here is very simple. Namely, the hypothesis is to evaluate the use of arsine (AsH3) against erythrocytes that have become infected with malaria parasites. Once malaria parasites have become established in erythrocytes, it is extremely difficult to displace them. Erythrocytes do not spontaneously undergo apoptosis, which is a mechanism that many types of cells would use to deny intracellular parasites an immune privileged domain. Moreover, antibiotics that are powerful and selective enough to defeat malaria parasites once they are established in the erythrocytes have not been found. Thus, approaches for selective hemolysis of malaria-infected erythrocytes and evicting the parasites may be attractive. It is not known, but should be easily determined experimentally, what the relative toxicity of arsine (AsH3) is to normal erythrocytes and malaria-infected erythrocytes. PMID- 15823694 TI - Asthma: a disuse contracture? AB - The incidence of asthma is increasing. There is no obvious explanation for this increase and current theory has no explanation for the occurrence of the disease in the non-allergic, the sudden onset of the asthma attack, the hyper-reactivity of the muscular wall, the association of the disease with obesity, or its precipitation by exercise. Biopsy studies have shown that the narrowing of air passages which characterises the disease is associated with thickening of their fibrous and muscular layers. As narrowing of air passages necessarily involves shortening of annular and helical components, this narrowing is in effect an annular contracture, analogous to those seen in underextended longitudinal structures such as muscles and tendons. The only common cause for such contractures is habitual underextension. As the only extending force in annular air passages is inspiration, this leads to the hypothesis that the basic cause of the disease is an insufficiency of aerobic exercise in childhood. Should the airways fail to develop their normal calibre, the narrowing will precipitate a sequence of events which can be predicted from the laws of physics. La Place's Law explains the instability of the muscle wall, the sudden onset of the attacks, reflecting episodes of critical collapse and the occurrence of the disease in the non-allergic. Bernoulli's Law explains the provocative effect of exercise and the postulated lack of exercise explains the increasing prevalence and the association with obesity. The hypothesis can be tested by comparative epidemiology. If it is correct, the disease should be preventable. PMID- 15823695 TI - Making cell-permeable antibodies (Transbody) through fusion of protein transduction domains (PTD) with single chain variable fragment (scFv) antibodies: potential advantages over antibodies expressed within the intracellular environment (Intrabody). AB - Over the past decade, there has been growing interest in the use of antibodies against intracellular targets. This is currently achieved through recombinant expression of the single chain variable fragment (scFv) antibody format within the cell, which is commonly referred to as an intrabody. This possesses a number of inherent advantages over RNA interference (iRNA). Firstly, the high specificity and affinity of intrabodies to target antigens is well-established, whereas iRNA has been frequently shown to exert multiple non-specific effects. Secondly, intrabodies being proteins possess a much longer active half-life compared to iRNA. Thirdly, when the active half-life of the intracellular target molecule is long, gene silencing through iRNA would be slow to yield any effect, whereas the effects of intrabody expression would be almost instantaneous. Lastly, it is possible to design intrabodies to block certain binding interactions of a particular target molecule, while sparing others. There is, however, various technical challenges faced with intrabody expression through the application of recombinant DNA technology. In particular, protein conformational folding and structural stability of the newly-synthesized intrabody within the cell is affected by reducing conditions of the intracellular environment. Also, there are overwhelming safety concerns surrounding the application of transfected recombinant DNA in human clinical therapy, which is required to achieve intrabody expression within the cell. Of particular concern are the various viral-based vectors that are commonly-used in genetic manipulation. A novel approach around these problems would be to look at the possibility of fusing protein transduction domains (PTD) to scFv antibodies, to create a 'cell-permeable' antibody or 'Transbody'. PTD are short peptide sequences that enable proteins to translocate across the cell membrane and be internalized within the cytosol, through atypical secretory and internalization pathways. There are a number of distinct advantages that a 'Transbody' would possess over conventional intrabodies expressed within the cell. For a start, 'correct' conformational folding and disulfide bond formation can take place prior to introduction into the target cell. More importantly, the use of cell-permeable antibodies or 'Transbodies' would avoid the overwhelming safety and ethical concerns surrounding the direct application of recombinant DNA technology in human clinical therapy, which is required for intrabody expression within the cell. 'Transbodies' introduced into the cell would possess only a limited active half-life, without resulting in any permanent genetic alteration. This would allay any safety concerns with regards to their application in human clinical therapy. PMID- 15823696 TI - Cardiac torsion and electromagnetic fields: the cardiac bioinformation hypothesis. AB - Although in physiology the heart is often referred to as a simple piston pump, there are in fact two additional features that are integral to cardiac physiology and function. First, the heart as it contracts in systole, also rotates and produces torsion due to the structure of the myocardium. Second, the heart produces a significant electromagnetic field with each contraction due to the coordinated depolarization of myocytes producing a current flow. Unlike the electrocardiogram, the magnetic field is not limited to volume conduction and extends outside the body. The therapeutic potential for interaction of this cardioelectromagnetic field both within and outside the body is largely unexplored. It is our hypothesis that the heart functions as a generator of bioinformation that is central to normative functioning of body. The source of this bioinformation is based on: (1) vortex blood flow in the left ventricle; (2) a cardiac electromagnetic field and both; (3) heart sounds; and (4) pulse pressure which produce frequency and amplitude information. Thus, there is a multidimensional role for the heart in physiology and biopsychosocial dynamics. Recognition of these cardiac properties may result in significant implications for new therapies for cardiovascular disease based on increasing cardiac energy efficiency (coherence) and bioinformation from the cardioelectromagnetic field. Research studies to test this hypothesis are suggested. PMID- 15823697 TI - Smoking promotes pathogenesis of aortic aneurysm through the 5-lipoxygenase pathway. AB - Smoking has been known as a risk factor for aortic aneurysm. 5-Lipoxygenase is the key enzyme in leukotriene biosynthesis and catalyzes initial steps in the conversion of arachidonic acid to these biologically active lipid mediators, which are known to exert proinflammatory effects in vivo. Smoking can induce 5 lipoxygenase expression in colon neoplasm, and may activate the 5-lipoxygenase pathway also in aortic tissue. 5-Lipoxygenase has a role in promoting the formation of aneurysms through potential plasma macrophage inflammatory protein 1alpha and -2 chemokine-dependent inflammatory circuits involving both myeloid and endothelial cells. Therefore, smoking may promote pathogenesis of aortic aneurysm through the 5-lipoxygenase pathway. PMID- 15823698 TI - A novel role for parkin in trauma-induced central nervous system secondary injury. AB - Recently, loss-of-function mutations of parkin have been identified as being causally related to autosomal recessive juvenile parkinsonism, the most common form of familial Parkinson's disease. In addition to functioning as an E3 ubiquitin ligase that facilitates the proteasomal degradation of proteins with abnormal conformations, parkin protects dopaminergic neurons from oxidative stress-mediated death by regulating mitochondrial function. Parkin is expressed throughout the brain in a variety of functional and neurochemical systems. We propose that parkin's role in protecting neurons from oxidative stress may extend beyond the nigrostriatal system to include neurons in other regions of the central nervous system. This is relevant for therapeutic strategies for brain and spinal cord injury because oxidative stress leading to lipid peroxidation and protein and nucleic acid oxidation is a significant cause of secondary injury and thus neuronal death following traumatic injuries to the central nervous system. A novel model system to verify the process of oxidative stress as a causative factor in trauma-induced secondary injury mechanisms would be to induce traumatic brain and spinal cord injury in parkin-null mice. This is expected to provide the proof-of-principle that a cascade of oxidative stress is a causal event leading to secondary neuronal injury, that parkin functions outside of the dopaminergic system to protect other neurons from oxidative stress, and that antioxidant pharmacotherapy is a rational therapeutic approach to decrease trauma-induced neuronal injury. PMID- 15823699 TI - Treatment of acute lymphocytic leukemia using zinc adjuvant with chemotherapy and radiation--a case history and hypothesis. AB - Low blood levels of zinc are often noted in acute lymphocytic leukemia (ALL), but zinc is not administered as part of any modern chemotherapy program in the treatment of ALL. Upon noting low blood levels of zinc in a 3-year-old 11.3 kg girl, zinc at the rate of 3.18 mg/kg body weight/day was administered from the start of chemotherapy through the full 3 years of maintenance therapy. Dosage was split with 18 mg given at breakfast and 18 mg zinc with supper. The result was a bone marrow remission from 95+% blast cells to an observed zero blast cell count in both hips within the first 14 days of treatment which never relapsed. In addition to the reduction of blast cells to an observed count of zero (not a single leukemic or normal blast), red blood cell production and other hemopoietic functions returned to normal at a clinically remarkable rate. There were no side effects from zinc or chemotherapy at any time, and zinc is hypothesized to have improved the patient's overall ability to withstand toxic effects of chemotherapy. This report identifies zinc treatment as being vital to rapid and permanent recovery from ALL. The extremely broad role of zinc in pre-leukemic adverse health conditions, viral, fungal and tumoral immunity, hemopoietics, cell growth, division and differentiation, genetics and chemotherapy interactions are considered. If a nutrient such as zinc could be shown to strengthen the function of chemotherapy and immune function, then it could be hypothesized that the relapse rate would be lessened since the relapse rate is related to both the rate at which a remission is obtained and the thoroughness of the elimination of leukemic blasts. Identical results also occurred in 13 other children with ALL whose parents chose to treat with zinc adjuvant. Since treatment with zinc and other identified deficient nutrients, particularly magnesium, did not appear injurious in ALL and they appear to be highly beneficial, controlled clinical studies of zinc (3.18 mg/kg body weight/day) with magnesium (8.0 mg/kg body weight/day) as adjuvants to chemotherapy in the treatment of childhood ALL are suggested. Treatment with zinc adjuvant is hypothesized to accelerate recovery from ALL, and in conjunction with chemotherapy, cure ALL. PMID- 15823700 TI - The chemical reactions in the human stomach and the relationship to metabolic disorders. AB - Acid hydrolysis of components from the diet in the stomach require the presence of an acid and a hydrolysing agent. The acid involved is hydrochloric acid. The present mechanism of hydrochloric acid production in the stomach is demonstrated to be incompatible with measured intracellular or intercellular concentrations of the relevant ions. An alternative set of chemical reactions whereby hydrochloric acid is formed in the stomach is presented. The hydrolysing agent is identified and a mechanism of transfer of chemical compounds into the metabolism is described. The hypothesis demonstrates that some of chemical compounds produced in the stomach can induce conditions such as asthma and that the conditions of osteoporosis and hemochromatosis can be linked to the function of the stomach. Possible treatments for these and other conditions such as stomach acidity and anaemia are proposed. PMID- 15823701 TI - The role of experimental myringosclerosis in interpretation of tympanograms and its possible clinical implications. AB - Myringosclerosis (MS) is hyalinization and calcification of the collagen layer in certain areas of the tympanic membrane (TM) and appears as white chalky patches in otomicroscopy. One of the most common sequela from the use of grommets is the development of MS and its rate in the tubed ear ranges between 44% and 54% during long term follow-up. Among tympanometric configurations, type As tympanograms may indicate ossicular fixation, tympanosclerosis or otitis media with effusion. In case of multiple pathologies (e.g., MS and otitis media with effusion), it is not possible to evaluate the effect of a specific pathology in the absence of other, with otomicroscopy and tympanometry. We believe that the movement of TM is hampered by lesions of sclerotic material, thus resulting in decrease of amplitude in tympanograms without any effusion in middle ear. Now, we propose an experimental model specific for creating MS solely on TM and for performing tympanometric measurements on this pure MS model without creating any pathology in the middle ear, to test in what proportion this specific pathology contributes to decrease of amplitude in tympanograms. A myringotomy in rat's TM admits ambient air into the middle ear cavity, resulting in a relatively hyperoxic condition. The result of a myringotomy therefore is an increased production of free oxygen radicals, initiating irreversible tissue damage involving fibrosis, hyalin degeneration and finally apopitosis as observed in MS. After the closure of perforations, tympanometric measurements can be made on this pure MS model. When evaluating a child for suspected otitis media with effusion, the results of the experimental model might have far-reaching clinical implications and might provide suitable target for prevention of unnecessary myringotomies especially in the pediatric age-group. PMID- 15823702 TI - Tranilast: a novel weapon against insulin resistance. AB - Oxidative stress and inflammatory cytokines such as monocyte chemoattractant protein 1 (MCP-1), TGF-beta, and IL-2 are supposed to play crucial roles in the pathogenesis of insulin resistance (IR). Tranilast is an anti-allergic drug which exerts anti-inflammatory and anti-angiogenesis effects through inhibition of expression of MCP-1, TGF-beta, and antigen-induced IL-2 lymphocyte responsiveness. It also possesses a certain antioxidant activity. Considering the above facts and in view of its safety, tranilast may prove invaluable in the treatment of IR. PMID- 15823703 TI - Melatonin seems to be a mediator that transfers the environmental stimuli to oocytes for inheritance of adaptive changes through epigenetic inheritance system. AB - Possibility of inheritance of epigenetic modifications have led us to consider that adaptive geographic variations in humans may result from interactions between environmental factors and epigenetic inheritance system. In this system melatonin seems to be a mediator that transfers the environmental stimuli to germ cells (oocytes). While environmental factors produce modifications in the body, they simultaneously induce epigenetic modifications in the oocytes with the help of melatonin, and these changes are inherited to offspring. In this way, adaptive changes could be passed on to the next generation. This kind of heritable long term changes is generally labeled biological adaptation. But, how can melatonin cause epigenetic changes in oocytes? We suggest that melatonin induces epigenetic modifications by affecting the nuclear melatonin receptors that can in turn change the superstructure of DNA. It was previously suggested that biological adaptation is limited to neural crest derivatives such as, craniofacial tissues, melanocytes, and structures related to stature, hair form and body proportions. Thus, inheritance of adaptive changes is possible only where environmental factors affect the neural crest derivatives, including the cells that produce the next generation. PMID- 15823704 TI - Uncertainties in the current understanding of gas exchange in spontaneous pneumothorax: effective lung ventilation may persist in a smaller-sized pneumothorax. AB - Clinical aspects of spontaneous pneumothorax (SP) are far more clear than some patophysiological issues. The exact mechanism that maintains adequate oxygenation in spontaneous pneumothorax of lesser size is still unclear. Experimental and rare studies in humans could not explicitely confirm whether it is hyperventilation of the nonaffected lung, still effective gas exchange within the affected lung, or hypoxic vasoconstriction. Similarly, it is unclear why the severity of dyspnoea sometimes differs between patients with the same size of SP. The idea that a certain degree of effective lung ventilation might exist in SP of lesser size was based on clinical observation of these patients on admission, on our measurements of pleural pressures and oxygenation in a group of patients with SP of different size and on rare experimental studies. Clinical observation that oxygenation was not significantly impaired in patients with SP of lesser size, without documented hyperventilation, served as a base for critical analysis of possible factors influencing oxygenation in SP of lesser size. Our hypothesis that pleural pressure swings in a partially collapsed lung, but still slightly expanding in inspiration, enable a certain degree of gas exchange, was confirmed both by several experimental studies and by our measurements. On the other hand, our clinical observation that patients with SP of greater size frequently differ in the severity of dyspnoea suggested the need of a more detailed analysis of the causes of hypoxaemia in these patients. The fact that hypoxaemia in these patients usually cannot be abolished by the existing hyperventilation, means that in SP of greater size, despite minimal lung volume, circulation in the pulmonary artery system still exists, causing right to left blood shunting. The fact that the severity of dyspnoea is not equal in all patients with complete SP means that hypoxic vasoconstriction exists only in some of them, following a still unknown pattern. Literature data and our measurements suggest that without further studies of hypoxic vasoconstriction in the acute phase of SP, the exact answer is not possible. PMID- 15823705 TI - Fludarabine phosphate may be useful in the treatment of graft-versus-host disease. AB - The increasing number of allogeneic stem cell transplantations has made the management of graft-versus-host disease (GVHD) a continuing problem for transplantation professionals. GVHD is a complicated disease the treatment of which requires an equally multifaceted approach. Despite therapeutic efforts to decrease its distressing and potentially lethal clinical manifestations, treatment is still not optimal. Fludarabine phosphate is a purine analogue, which is known to cause immunosuppression and long-lasting T-cell lymphopenia it is commonly employed in the therapy of hematological malignancies and non myeloablative stem cell transplantation conditioning regimens. Myelosuppression, especially leuko- and lymphopenia is the major dose-limiting toxicity of fludarabine. However, a prolonged reduction in CD4+ T-cell count may be a desired effect for the treatment of GVHD. Clinical observations, preclinical data on the management of GVHD and well-known immunosuppressive properties suggest that fludarabine should be tested in clinical grounds for GVHD prophylaxis and treatment. PMID- 15823706 TI - Hypothesis: a motor neuron toxin produced by a clostridial species residing in gut causes ALS. AB - We hypothesize that a yet-to-be-identified motor neuron toxin produced by a clostridial species causes sporadic amyotrophic lateral sclerosis (ALS) in susceptible individuals. This clostridial species would reside undetected in the gut and chronically produce a toxin that targets the motor system, like the tetanus and botulinum toxins. After gaining access to the lower motor neuron, the toxin would be transported back to the cell body, as occurs with the tetanus toxin, and destroy the lower motor neuron - the essential feature of ALS. Again like the tetanus toxin, some of the toxin would cross to neighboring cells and to the upper motor neuron and similarly destroy these motor neurons. Weakness would relentlessly progress until not enough motor neurons remained to sustain life. If this hypothesis were correct, treatment with appropriate antibiotics or antitoxins might slow or halt progression of disease, and immunization might prevent disease. PMID- 15823707 TI - A hypothesis matrix for studying biomechanical factors associated with the initiation and progression of posttraumatic osteoarthritis. AB - In this paper the current theories on how osteoarthritis (OA) may be initiated and progressed is described. This is done in relation to the biomechanical events that would predispose the joint to the degenerative process, as well as further progression of the process within an 'OA' cycle. The relationship between the types of loading to the type of joint damage that occurs is discussed. Subsequently the influence on the rate at which OA progresses from the trauma, is presented within a hypotheses matrix. For the type of tissue damage, four phases are distinguished, phase I: superficial cells or matrix only, phase II: deeper chondral region, phase III: the tidemark or calcified region, and phase IV: subchondral bone region. The biomechanical event (A) is stipulated as having a possibility of six outcomes. (A3) is the direct damage to the calcified cartilage near the tidemark that leads most rapidly into the cycle for OA to develop and progress. Another three outcomes (A1, A2 and A4) involve damage to regions other than the calcified cartilage near the tidemark. These three outcomes involve the cells or matrix, chondral or subchondral regions. It is hypothesised that damage involved in one of these three outcomes results in all likelihood to a new level of joint deficiency or vulnerability. This new predisposition could lead to A3 type outcome and directly into the OA progression cycle or result in more A1, A2 or A4 type outcomes which remains out of the OA cycle. The biomechanical events are therefore used to predict the risk of mechanically driven OA and the rapidity in which it progresses in relation to joint loading. PMID- 15823708 TI - Selenium supplementation may increase the efficacy of cetuximab in metastatic colorectal cancer patients. AB - Recently, cyclooxygenase-2 (COX-2) inhibitor therapy has emerged as a possible new approach to the prevention and treatment of colorectal cancer (CRC). The COX enzymes (COX-1 and COX-2) are key enzymes of prostaglandin (PG) biosynthesis and are overexpressed in approximately 80% of human CRCs. Presumably, bioactive lipid products of COX, such as PGE(2), are responsible for some of the pro-neoplastic effects mediated by this enzyme. The early effects of COX-2-derived PGE(2) are in part mediated by the epidermal growth factor receptor (EGFR). Selenomethionine decreases COX-2 protein and PGE(2) levels. Cetuximab is a chimeric IgG1 monoclonal antibody that binds to EGFR with high specificity thus blocking ligand induced phosphorylation of EGFR. Cetuximab has clinically significant activity when given alone or in combination with irinotecan in patients with irinotecan refractory CRC. We suggest that selenium supplementation by decreasing the COX-2 protein and PGE-2 levels in cancer cells may increase efficacy of cetuximab in advanced CRC patients. PMID- 15823709 TI - Reconstruction of the anterior cruciate ligament with a patella-tendon-bone graft may lead to a permanent loss of bone mineral content due to decreased patellar tendon stiffness. AB - Immobilisation induces bone loss. Evidence from studies in animals and healthy humans that were immobilised for a limited time indicates that, in general, bone mass may be restored even in adults. Following conservative management of partial tears of the anterior cruciate ligament (ACL), bone loss is often negligible (2 3%). After surgical reconstruction, however, there is greater bone loss (15-20%), with little or no recovery. Bones adapt to the stresses they experience. Also, the largest forces in the musculoskeletal system arise from muscle pull. Tendons transmit these forces. Many surgical techniques for ACL reconstruction use autologous tendon grafts. We hypothesise that tissue harvesting causes weakening of the formerly intact tendon, which, in turn, leads to reduced muscle pull and subsequent bone loss in those parts of the bone that are loaded by the tendon. If our hypothesis holds true, it may change patients' and surgeons' choice of management. Clinical follow-up should assess the functional result with greater scrutiny, possibly including the assessment of bone mineral content. This may be particularly important since there is accumulating evidence that a decrease in bone mineral density (BMD) preceedes, and hence may be a cause of, osteoarthritis. PMID- 15823710 TI - Lysosomal and prostasomal hydrolytic enzymes and redox processes and initiation of prostate cancer. AB - Lysosomes, lysosomal enzymes and oxidant processes are known to be involved in cancer processes. The prostasomes contain proteins and enzymes that would constitute pathways for the hydrolysis of proteins and peptides. However, integrated biochemical and cell biology studies are necessary to understand how lysosomal enzymes and prostasomal enzymes combined with oxidant processes could initiate cancer. Most prostate cancer is likely to be initiated in the prostate duct system. The lysosomal enzymes acid phosphatase and glucosidase and prostasomal proteins and enzymes are found in human semen and therefore have come through prostate ducts. The hypothesis presented here is that the lysosomal enzymes and prostasomes are exocytosed from prostate cells into the duct system of the prostate where their hydrolytic enzymes and oxidative processes, for example, the iron from the iron-sulfur clusters of the prostasomal dehydrogenases, damage proteins and other components of cells leading to the initiation of cancer. Risk factors for prostate cancer are known to initiate activity of lysosomal enzymes and could initiate activity of prostasomal enzymes. These risk factors include: ionizing radiation, oxidative stress, environmental toxicants and dietary components including those with high fat content. Other dietary components in fruits and vegetables protect against prostate cancer and can be hypothesized as decreasing cellular output of lysosomal or protasomal enzymes or inhibiting lysosomal and prostasomal enzymes in the duct system. Measurements of multiple lysosomal and prostasomal enzyme activities and their biochemical pathways are vital to the understanding of protectors to inhibit lysosomal or prostasomal enzyme activities that might be leading to prostate cancer. Inhibitors of lysosomal and prostasomal enzymes can be investigated in cellular and biochemical systems, and these inhibitors could be used to control these enzyme activities in vivo. In situ enzyme analyses including substrates producing fluorescent products are applicable. Screening assays could be developed to detect in vivo lysosomal and prostasomal enzyme activities in semen. Lysosomal enzyme activities may be precursors to the onset of other kinds of cancer with other similar non-invasive screening techniques possible. Present knowledge encompasses mobilization of sperm when prostasomes bind to sperm in semen. A further hypothesis of this study projects that prostasomal dehydrogenases and their NADH products initiate the formation of ATP in the sperm mitochondria which activates flagellar movement. This overall hypothesis suggests protection against prostate cancer by inhibitors of lipid peroxidation including the dietary antioxidants selenium, vitamin E and lycopene and also cysteine glutathione. PMID- 15823711 TI - Significant reduction of the risk of venous thromboembolism in all long-term immobile patients a few months after the onset of immobility. AB - Prophylactic anticoagulation is a standard practice in patients with sudden lower limbs paralysis. Thromboprophylaxis is usually continued until the patient regains independent mobility. The duration of anticoagulation in long-term immobile patients is unknown. Spinal cord injury patients are the only population that was comprehensively studied and prophylactic anticoagulation is discontinued after 4 months as the risk of venous thromboembolism drops dramatically after 3-4 months. Development of muscle spasticity has been traditionally considered to be the reason for this low risk as lower limbs spasticity/spasms might be able to improve the calf muscle pump action. We are presenting the evidence from physiological studies of the lower limbs vascular system that cast doubt over this explanation and present an alternative hypothesis backed by several clinical circumstantial evidence suggesting that the vascular changes following long term lower limbs inactivity which are universal to all immobile patients is probably the main protecting factor. We suggest that prophylactic anticoagulation is necessary only on the first 4 months following the acute onset of immobility in all neurologically impaired immobile patients regardless of their muscle tone state. PMID- 15823712 TI - Motion sickness is linked to nystagmus-related trigeminal brain stem input: a new hypothesis. AB - Motion sickness is a common and distressing but poorly understood syndrome associated with nausea/vomiting and autonomic nervous system accompaniments that develops in the air or space as well as on sea or land. A bidirectional aetiologic link prevails between migraine and motion-sickness. Motion sickness provokes jerk nystagmus induced by both optokinetic and vestibular stimulation. Fixation of gaze or closure of eyes generally prevents motion sickness while vestibular otolithic function is eliminated in microgravity of space, indicating a predominant pathogenetic role for visuo-sensory input. Scopolamine, dimenhydrinate, and promethazine reduce motion-related nystagmus. Contraction of extraocular muscles generates proprioceptive neural traffic and can provoke an ocular hypertensive response. It is proposed that repetitive contractions of the extraocular muscles during motion-related jerk nystagmus rapidly augment brain stem afferent input by increasing proprioceptive neural traffic through connections of the oculomotor nerves with the ophthalmic nerve in the lateral wall of the cavernous sinus as well as by raising the intraocular pressure thereby stimulating anterior segment ocular trigeminal nerve fibers. This verifiable hypothesis defines the pathophysiological basis of individual susceptibility to motion sickness, elucidates the preventive mechanism of gaze fixation or ocular closure, advances the aetiologic link between MS and migraine, rationalizes the mechanism of known preventive drugs, and explores new therapeutic possibilities. PMID- 15823713 TI - Selective reduction of estrogen receptor (ER) positive breast cancer occurrence by estrogen receptor modulators supports etiological distinction between ER positive and ER negative breast cancers. AB - Reports that selective estrogen receptor modulators (SERMs) reduce occurrence of only estrogen receptor (ER) positive tumors strongly support the etiological distinction between ER positive and ER negative breast cancers. Based on these evidences three concepts are described: Concept I. The occurrence of ER negative tumor cells might be a consequence of the clonal selection among tumor cells. This would lead to mosaicism in the ER expression. If ER negative cells become the most prevalent clone, the patient will be diagnosed to have an ER negative breast cancer. Since all cancers start as ER positive, SERMs should equally prevent occurrence of ER positive and ER negative breast cancers, but this prediction is evidently wrong. Concept II. Mammary ducts normally contain ER positive and ER negative cells, both prone to malignancy. Cancer occurrence in ductal cells that normally lack ER would be unrelated to estrogen exposure or SERMs protection. Estrogen and SERMs can influence cancer occurrence only in ER positive ductal cells. The main drawback is that this concept does not predict occurrence of mosaicism in ER expression among tumor cells. Unified Concept I and II. To overcome limitations of described concept a unified concept is presented. Cancers from ER positive ductal cells start as pure ER positive tumors and those from ER negative ductal cells as pure ER negative tumors. During the preclinical phase, in some ER positive tumors, clonal selection introduces ER negative clones. These tumors become mosaic in the cellular ER expression, some of them predominantly ER positive other ER negative. Estrogen deprivation, or SERMs can help mostly to patients with pure ER positive, or mosaic ER positive tumors. Since the dominant metastatic clone can have different ER status from the primary breast tumor, both surprising successes and failures of endocrine therapy can be expected in tumors with mosaic ER expression. PMID- 15823714 TI - Primary nocturnal enuresis: omega-3 fatty acids may be of therapeutic value. AB - Primary nocturnal enuresis (PNE), or bed-wetting, is a distressing urinary condition which can persist through childhood and beyond. Altered prostaglandin and nitric oxide production have been observed in children with PNE, and prostaglandin inhibitors are known to be of therapeutic value. Omega-3 fatty acids have the potential to influence the symptoms of PNE by inhibition of prostaglandin and renal nitric oxide production. In addition, children with PNE have an inappropriate startle response and an apparent maturational delay of the central nervous system. Research clearly shows that omega-3 fatty acids play a critical role in the development and function of the central nervous system. It is our contention that inadequate omega-3 intake may play a role in the lack of inhibitory input to the startle and micturition centers in PNE. PMID- 15823715 TI - Novel hypothesis for the cause of panic disorder via the neuroepithelial bodies in the lung. AB - Panic disorder (PD) is a complex condition that is further complicated by its numerous inducers, which include hypercapnia, hypoxia, sodium lactate, caffeine and cholecystokinin. It seems unlikely that there are specific suffocation receptors for each of these inducers in the brain. The pulmonary neuroepithelial bodies (NEBs), which are situated at the bifurcation point of the small bronchi, act as storage cells for 5-hydroxytryptamine (5-HT) and sensors for suffocation. If we suppose that PD might represent an inflammation of the NEBs, bradykinin (BK) which augments the airway hyper-response to diverse indcers might cause these cells to release 5-HT along with peptides and panneuroendcrine markers from their dence-core secretory granules. It was revealed that BK with 5-HT could cross the blood-brain barrier (BBB). When 5-HT released from these cells along with BK cross the BBB, the release of 5-HT at the axonal terminals in the serotonergic neurons in the brain will be inhibited, since the 5-HT1 autoreceptor have a higher affinity for 5-HT than do the 5-HT2 receptors. The inhibition of 5 HT at the axonal terminal causes to suppress the periaqueductal gray matter, which inhibits flight reactions to impending danger, pain or asphyxia. In short, this serotonergic situation might bring about PD. According to this theory, the type of inducer that the PD patient is exposed to is unimportant as long as it stimulates the NEBs, and through the effect of 5-HT and BK, PD would be revaluated as a somatic disease that directly and reversibly affects the brain. PMID- 15823716 TI - The influence of weather and environment on pulmonary embolism: pollutants and fossil fuels. AB - Previous publications have highlighted seasonal variations in the incidence of thrombosis and pulmonary embolism, and that weather patterns can influence these. While medical risk factors for pulmonary thrombo-embolism such as age, obesity, hypercoagulable states, cancer, previous thrombo-embolism, immobility, limb paralysis, surgery, major illness, trauma, hypotension, tachypnoea and right ventricular hypokinesis are not directly implicated regarding environmental factors such as weather, they could be influenced indirectly by these. This would be especially relevant in polluted areas that are associated with a higher pulmonary embolism risk. Routine nuclear medicine lung ventilation/perfusion studies (V/Q scans) of 2071 adult patients referred to the nuclear medicine department of the Royal Surrey County Hospital in Guildford, UK, between January 1998 and October 2002 were reviewed and 316 of these patients were classified as positive for pulmonary embolism with high probability scan on PIOPED criteria. The occurrence of positive scans was compared to environmental factors such as temperature, humidity, vapour pressure, air pressure and rainfall. Multiple linear regression was used to establish the significance of these relations. The incidence of pulmonary embolism was positively related to vapour pressure and rainfall. The most significant relation was to vapour pressure (p=0.010) while rainfall was less significant (p=0.017). There was no significant relation between pulmonary embolism and air pressure, humidity or temperature. It is postulated that rainfall and water vapour may be contributary factors in thrombosis and pulmonary embolism by way of pollutants that are carried as condensation nuclei in micro-droplets of water. In particular, fossil fuel pollutants are implicated as these condensation nuclei. Pollutants may be inhaled by populations exposed to windborne vapour droplets in cities or airports. Polluted vapour droplets may be absorbed by the lung to hasten coagulation cascades in the blood. This may lead to thrombosis and increased pulmonary embolism under high vapour pressure conditions. With combined factors such as pre existing ill health or immobility on long flights, the risk of thrombosis and consequent embolism might increase substantially. PMID- 15823717 TI - Met72Thr polymorphism of pigment epithelium-derived factor gene and susceptibility to age-related macular degeneration. AB - Age-related macular degeneration (ARMD) is the most common cause of acquired blindness among the people of occupational age. Although the pathogenesis of ARMD is not fully understood, several studies suggest a possible contribution of a genetic factor in the development and progression of ARMD. Pigment epithelium derived factor (PEDF), a glycoprotein that belongs to the superfamily of serine protease inhibitors, was first purified from the conditioned media of human retinal pigment epithelial cells as a factor with potent neuronal differentiating activity in human retinoblastoma cells. Recently, PEDF has been shown to be a highly effective inhibitor of angiogenesis in cell culture and animal models. In addition, PEDF has been found in the vitreous, and its levels were decreased in angiogenic eye diseases, thus suggesting that a loss of PEDF in the eye is functionally important in the pathogenesis of ARMD. A functional amino acid change, a methionine to threonine polymorphism (Met72Thr polymorphism) at codon 72 in exon 3 (T/C polymorphism) of the PEDF gene, that results in the formation of BsstSI restriction site, has recently been identified. Since it is well known that a single nucleotide polymorphism and resultant amino acid change often alters the activity or expression level of the target protein, we would like to propose here a novel hypothesis that the Met72Thr polymorphism (T/C polymorphism) of PEDF gene may be a genetic marker for ARMD. Are genotype and allele frequencies of the Met72Thr polymorphism (T/C polymorphism) different between the patients with or without ARMD? Is this polymorphism associated with disease severity and progression? If the answer is yes, does this Met72Thr polymorphism regulate the vitreous levels of PEDF? These clinical studies could provide us with information whether this genetic variant of the PEDF gene could present an attractive candidate susceptibility gene for ARMD. PMID- 15823718 TI - Serum or cerebrospinal fluid levels of glyceraldehyde-derived advanced glycation end products (AGEs) may be a promising biomarker for early detection of Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common cause of dementia in developed countries. AD is characterized pathologically by the presence of senile plaques and neurofibrillary tangles (NFTs), the major constituents of which are the amyloid beta protein (Abeta) and tau protein, respectively. Several epidemiological studies have reported moderately increased risks of AD in diabetic patients compared with general population. In diabetes mellitus, the formation and accumulation of advanced glycation end products (AGEs) progress. Recent understandings of this process have confirmed that AGEs - their receptor (RAGE) interactions may play a role in the pathogenesis of diabetic vascular complications and neurodegenerative disorders including AD. Indeed, it has been demonstrated that AGEs can be identified immunohistochemically to be present in both senile plaques and NFTs from patients with AD. Glycation of Abeta markedly enhances its aggregation in vitro, and the glycation of tau, in addition to hyperphosphorylation, appears to enhance the formation of paired helical filaments. Further, RAGE has been found a specific cell surface receptor for Abeta peptite, thus eliciting neuronal cell perturbation. The active participation of RAGE in the pathogenesis of AD has also been confirmed in RAGE overexpressed transgenic mice. Moreover, we have recently found that glyceraldehyde-derived AGEs, one of the representative ligands for RAGE, exerted cytopathic effects on cultured neuronal cells and that neurotoxic effect of diabetic serum was completely blocked by neutralizing antibodies against glyceraldehydes-derived AGEs. These observations led us to hypothesize that serum or cerebrospinal fluid (CSF) levels of glyceraldehyde-derived AGEs could become a promising biomarker for early detection of AD. We also would like to propose the possible ways of testing our hypothesis. Are the concentrations of glyceraldehyde derived AGEs in serum or CSF elevated early in the course of dementia? Are these levels correlated with disease severity and progression, especially in patients with diabetes? These clinical studies clarify whether use of serum or CSF levels of glyceraldehyde-derived AGEs as a biomarker for AD might enable more effective diagnosis and treatment of patients with this devastating disorder. PMID- 15823719 TI - Possible participation of advanced glycation end products in the pathogenesis of colorectal cancer in diabetic patients. AB - Colorectal cancer is a major public health problem, being the second most common cause of cancer in developed countries. Several epidemiological studies have reported moderately increased risks of colorectal cancer in diabetic patients compared with general population. However, the underlying molecular link between diabetes and colorectal cancer remains to be elucidated. In diabetes mellitus, the formation and accumulation of advanced glycation end products (AGEs) progress. There is a growing body of evidence to show that AGEs-their receptor (RAGE) interactions are involved in the development of atherosclerosis and diabetic microangiopathy. AGEs-RAGE interactions stimulated the growth of human pancreatic cancer cells through the autocrine induction of platelet-derived growth factor-B. Furthermore, we have recently found that AGEs stimulated the growth and migration of cultured human melanoma cells and that anti-RAGE antibodies inhibited tumor formation and lung metastasis of melanoma cell xenografts and subsequently improved survival in athymic mice. These observations let us to hypothesize that AGEs could explain the molecular link between diabetes and colorectal cancer. In this paper, we would like to propose the possible ways of testing our hypotheses. Is elevation of serum AGE levels a risk factor for colorectal cancer in patients with diabetes? Does treatment with metformin, which has a potential effect on the inhibition of glycation reactions in vivo, decrease the risk for colorecetal cancer in diabetic patients? If the answer is yes, is this beneficial effect of metformin superior to that of other anti-diabetic agents with equihypoglycemic properties? Does treatment with pyridoxamine, a post Amadori inhibitor (so-called Amadorins) of AGE formation, reduce the risk for colorectal cancer as well? Furthermore, are increased levels of AGEs and RAGE in colorectal cancer associated with poor prognosis in patients with diabetes? These clinical studies could clarify whether the AGEs-RAGE interactions serve as a causal link between diabetes and colorectal cancer. PMID- 15823720 TI - AMPK activation as a strategy for reversing the endothelial lipotoxicity underlying the increased vascular risk associated with insulin resistance syndrome. AB - The endotheliopathy associated with insulin resistance syndrome appears to result largely from excessive free fatty acid (FFA) exposure that boosts endothelial production of diacylglycerol, thereby activating protein kinase C. This endothelial "lipotoxicity" can be alleviated by very-low-fat diets and by appropriate weight loss. In addition, pharmacological activation of endothelial AMP-activated kinase (AMPK), as with the drug metformin, has the potential to decrease the FFA content of endothelial cells by stimulating fat oxidation; AMPK may also suppress endothelial de novo synthesis of diacylglycerol by inhibiting glycerol-3-phosphate acyltransferase. These considerations may rationalize the superior impact of metformin therapy on the macrovascular health of diabetics. More generally, metformin - or, preferably, better tolerated activators of AMPK - may have considerable potential for promoting vascular health in the large proportion of the adult population afflicted with insulin resistance syndrome. PMID- 15823721 TI - High mitochondrial redox potential may promote induction and activation of UCP2 in hepatocytes during hepatothermic therapy. AB - Although uncoupling protein-1 is a key mediator of thermogenesis in activated brown fat, the more recently characterized uncoupling proteins-2 and -3 do not appear to influence basal metabolism, but rather may function to diminish excessive mitochondrial superoxide production when mitochondrial redox potential is high. Under these circumstances, superoxide within the mitochondrial matrix directly activates uncoupling protein-2 (UCP2), and may also promote induction of this protein. Normal healthy hepatocytes do not express UCP2, but this protein is induced in hepatocytes that are steatotic or that are treated with agents that boost superoxide production. It is proposed that induction and activation of UCP2 may play a role in the thermogenesis evoked by hepatothermic therapy, a strategy designed to decrease body fat by maximizing hepatic fatty acid oxidation. Under these conditions, high mitochondrial redox potential would be expected, and induction of UCP2's uncoupling activity would represent a homeostatically appropriate antioxidant response. PMID- 15823722 TI - Chromium picolinate may favorably influence the vascular risk associated with smoking by combating cortisol-induced insulin resistance. AB - Smoking promotes insulin resistance and other features - excepting hypertension - of the insulin resistance syndrome; these effects appear to reflect chronic nicotine exposure. The adverse impact of smoking on insulin function may be a prominent mediator of the excess vascular risk associated with smoking. Although the mechanistic basis of nicotine-induced insulin resistance remains to be clarified, increased secretion of ACTH and cortisol seems likely to play an important role in this regard. It is therefore intriguing to note that supplemental chromium picolinate has been reported to have a rapid and substantial favorable impact on glycemic control in patients with corticosteroid induced diabetes. In dexamethasone-treated rats, high doses of this compound have been shown to markedly improve insulin sensitivity. Thus, if these initial reports are confirmable, ample intakes of bioactive chromium may have the potential to reverse the negative influence of corticosteroids on insulin signaling. Such an effect might have utility not only for smokers, but for others in whom chronic up-regulation of the hypothalamic-pituitary-adrenal axis compromises vascular health. The impact of chromium picolinate on smoking- or nicotine-induced insulin resistance merits study. PMID- 15823723 TI - The empirical relationship between longevity and physical stature may be obscured by unobserved genetic diversity in the optimal metabolic rate. AB - It is hypothesized that an inheritable "optimal" metabolic rate is associated with the lowest attainable mortality risk, greatest attainable height (in youth) and a desirable body weight. Positive deviations from this rate are reflected in overweight and higher mortality, negative deviations in shorter stature (stunting), low body mass index (BMI), and higher mortality. In a heterogeneous population the optimal rate may differ among genotypes, and those with a higher optimal rate are the ones who can translate more energy intake into growth in height such that under optimal net-nutritional intake, they are able to reach a greater final adult height with a lower body mass index than genotypes with a lower optimal rate. Hence, the optimal height and weight - in terms of longevity may differ substantially among genotypes, so that an empirical estimation of the relationship may be blurred in a heterogeneous society. Conversely, the "true" association (within genotypes) may be stronger than the measured one, which may have some implications for the contribution of the secular increase in human height to the rise in life expectancy or the optimal BMI in persons who experienced stunting in youth. PMID- 15823724 TI - Assessing the efficacy of antidepressants: the transactional paradigm. AB - Extant pharmacological literature seems to indicate that antidepressants are the most effective treatment for depression. However, there are flaws in the traditional paradigm for evaluating the efficacy of antidepressants. The traditional paradigm in pharmacology has adopted a biologically monistic causal structure, suggesting that the efficacy of antidepressants resides exclusively in biological pathways. The current paradigm in pharmacology has ignored the potential impact of psychological factors associated with the efficacy of antidepressants. This has occurred because psychological effects are seen as being a "paradigmatic anomaly," phenomena that are at odds with the traditional paradigm. Thus, the psychological factors associated with the administration of antidepressants have collectively been labeled as a "placebo effect" and discounted as a potential treatment pathway. However, empirical evidence suggests that the psychological factors, such as the formation of patient expectancies and a therapeutic alliance between physician and patient, increase the efficacy of antidepressants. Consequently, the current paper proposes the adoption of a transactional model that reflects the reciprocal interaction between psychological and biological factors associated with the efficacy of antidepressants. PMID- 15823725 TI - Is atrial fibrillation an inflammatory disease? PMID- 15823726 TI - A rationale for the development of resistance to adriamycin (doxorubicin). PMID- 15823727 TI - Bisphosphonates may inhibit development of atherosclerosis formation through its bactericidal effect on nanobacteria. PMID- 15823728 TI - Mathematical model for polygenic diseases. PMID- 15823729 TI - Denervation-reinnervation--hypothesis well-founded but not sufficient. PMID- 15823730 TI - Can the high nuclear to cytoplasmic ratio of human embryonic stem cells make them more vulnerable to physical stress encountered with bulk-passage protocols? PMID- 15823731 TI - DNA methylation and the central dogma of molecular biology. PMID- 15823732 TI - Association of skin rash and tumor response to HER1/EGFR inhibition: does HER1 stimulated tumor growth depend on circulatory instead of paracrine ligands? PMID- 15823733 TI - The anti-inflammatory and antioxidant effects of long-chain n-3 fatty acids or oil-rich fish may favorably affect atrial remodeling in atrial fibrillation. PMID- 15823734 TI - The potential utility of sodium valproate as a novel addition to anti-multiple myeloma armamentarium. PMID- 15823735 TI - Possible metabolism--body weight effect on prolongation and reduction of the pregnancy duration. PMID- 15823736 TI - Therapy of newly emerging mutant viral disorders and role in bioterrorism. PMID- 15823737 TI - Short small bowel patients are permanently disabled due to unrecognised starvation. PMID- 15823739 TI - Laboratory development and field demonstration of self-sealing/self-healing landfill liner. AB - The self-sealing/self-healing (SS/SH) barrier concept is based on the principle that two or more parent materials placed in vertical or horizontal layers will react at their interfaces to form insoluble reaction products. These products constitute a seamless impermeable seal, which is resistant to the transmission of leachate and contaminants. A SS/SH liner formulation was developed in the laboratory and demonstrated at the Sudokwon landfill site in South Korea. Laboratory testing results indicated that a seal with a hydraulic conductivity less than 10(-9) m/s formed after two to four weeks of curing at room temperature, and the seal healed itself after it was fractured. The use of the soil from the Sudokwon landfill site instead of sand as the matrix of the parent materials in the SS/SH liner retarded the sealing and healing of the seal, but did not show an obvious effect on the overall sealing and healing capacity of the seal at early stages. The construction and installation of the field demonstration SS/SH liner were carried out in the same way as for a soil cement liner. The quality of the liner was ensured by the enforcement of quality analysis/quality control procedures during installation. A single sealed ring infiltration test was performed on the field demonstration liner 36 days after the installation was completed. The measurement of water infiltration rate indicated that the liner healed after it was fractured. However, the long-term sealing and healing capacity needs to be further investigated. PMID- 15823740 TI - The production, use and quality of sewage sludge in Denmark. AB - In Denmark, the production of municipal sewage sludge decreased from approximately 170,000 ton d.m. in 1994 to 140,000 ton d.m. in 2002. The sludge is handled and treated in a number of ways. The quality of Danish sludge has steadily improved since the middle of the 1980s, when the first set of quality criteria for heavy metals was introduced. In 1997, cut-off criteria for the organic pollutants, LAS, DEHP, nonylphenol and PAHs were introduced. Effective control from authorities, voluntary phasing out agreements with industry, improved source identification tools, better handling and after-care methods have in combination with higher waste duties led to a significant reduction in the sludge level of especially cadmium, mercury, chromium, LAS and nonylphenol. The increased quality demand has, nevertheless, also led to a minor reduction in the use of sewage sludge as organic fertiliser on agricultural land. PMID- 15823741 TI - Elutriation characteristics of fine particles from bubbling fluidized bed incineration for sludge cake treatment. AB - In this study, measurements of elutriation rate were carried out in a bench scale bubbling fluidized bed incinerator, which was used to combust sludge cake. The particle size distribution and ignition loss were analyzed to study the elutriation characteristics of bubbling fluidized bed incineration. Drawn from the experimental data, the elutriation rate constant K(i)* for fine particles were obtained and correlated with parameters. It was found that most of the solid particles (about 95%) elutriated came from the fluidized medium (inorganic matters), but few came from unburned carbon particles or soot (about 5%). Finally, this paper lists a comparison of K(i)* between this study and the published prediction equations derived or studied in non-incineration modes of fluidized bed. A new and modified correlation is proposed here to estimate the elutriation rate of fine particles emitted from a bubbling fluidized bed incinerator. Primary operation variables (superficial gas velocity and incineration temperature) affecting the elutriation rate are also discussed in the paper. PMID- 15823742 TI - LIFAC ash--strategies for management. AB - LIFAC is a more recent addition to flue gas desulphurization methods for reducing sulphur emissions during coal combustion for the production of electricity. Ashes from the combustion of a low-sulphur lignite coal using LIFAC technology were used to evaluate different ash management strategies. The ashes, as produced and after treatment by the CERCHAR hydration process, were examined for their disposal characteristics and their utilization potential in concrete. They were also evaluated as underground disposal material using the AWDS process. PMID- 15823743 TI - Laboratory studies of the remediation of polycyclic aromatic hydrocarbon contaminated soil by in-vessel composting. AB - The biodegradation of 16 polycyclic aromatic hydrocarbons (PAHs), listed as priority pollutants by the USEPA, present in a coal-tar-contaminated soil from a former manufactured gas plant site was investigated using laboratory-scale in vessel composting reactors to determine the suitability of this approach as a bioremediation technology. Preliminary investigations were conducted over 16 weeks to determine the optimum soil composting temperature (38, 55 and 70 degrees C). Three tests were performed; firstly, soil was composted with green-waste, with a moisture content of 60%. Secondly, microbial activity was HgCl2-inhibited in the soil green-waste mixture with a moisture content of 60%, to evaluate abiotic losses, while in the third experiment only soil was incubated at the three different temperatures. PAHs and microbial populations were monitored. PAHs were lost from all treatments with 38 degrees C being the optimum temperature for both PAH removal and microbial activity. Calculated activation energy values (E(a)) for total PAHs suggested that the main loss mechanism in the soil-green waste reactors was biological, whereas in the soil reactors it was chemical. Total PAH losses in the soil-green waste composting mixtures were by pseudo-first order kinetics at 38 degrees C (k = 0.013 day(-1), R2 = 0.95), 55 degrees C (k = 0.010 day(-1), R2 = 0.76) and at 70 degrees C (k = 0.009 day(-1), R2 = 0.73). PMID- 15823744 TI - Carbonation of MSWI-bottom ash to decrease heavy metal leaching, in view of recycling. AB - The 0.1-2 mm fraction of a MSWI-bottom ash cannot be used as granular construction material because leaching of Cu exceeds Flemish limit values. In addition, leaching of Ba, Mo and Sb exceeds informal limit values. Leaching characteristics thus need to be improved. Carbonation was the chosen treatment method and this was performed by placing samples in a CO2 chamber. The CO2 percentage and the temperature of the chamber atmosphere, as well as the initial humidity of the samples, were varied to optimize carbonation parameters. Metal leaching was tested with the EN 12457 extraction test. Carbonation decreased Cu leaching from 3.3 to 1.0 mg/kg, but not yet to below the official limit value of 0.5 mg/kg. Leaching of Mo and Sb remained fairly constant or even increased after carbonation, but their limit values are only informal. Ba leaching decreased to below the informal limit value. Carbonation also caused Cr leaching to increase, in some cases to above the official limit value. Of the tested parameters, a CO2 percentage of 10% and a carbonation temperature of 50 degrees C in the atmosphere, together with ash humidity between 13% and 25% appeared to give the best leaching results. The main carbonation reactions took place within the first 24 h. PMID- 15823745 TI - Behavior of metals in ash melting and gasification-melting of municipal solid waste (MSW). AB - In this study, metal behavior in ash-melting and municipal solid waste (MSW) gasification-melting facilities were investigated. Eight ash-melting and three MSW gasification-melting facilities with a variety of melting processes and feedstocks were selected. From each facility, melting furnace fly ash (MFA) and molten slag were sampled, and feedstock of the ash-melting processes was also taken. For the ash melting process, the generation rate of MFA was well correlated with the ratio of incineration fly ash (IFA) in feedstock, and this was because MFA was formed mostly by mass transfer from IFA and a limited amount from bottom ash (BA). Distribution ratios of metal elements to MFA were generally determined by volatility of the metal element, but chlorine content in feedstock had a significant effect on Cu and a marginal effect on Pb. Distribution ratio of Zn to MFA was influenced by the oxidizing atmosphere in the furnace. High MFA generation and distribution ratio of non-volatile metals to MFA in gasification melting facilities was probably caused by carry-over of fine particles to the air pollution control system due to large gas volume. Finally, dilution effect was shown to have a significant effect on metal concentration in MFA. PMID- 15823746 TI - Emissions from small-scale energy production using co-combustion of biofuel and the dry fraction of household waste. AB - In sparsely populated rural areas, recycling of household waste might not always be the most environmentally advantageous solution due to the total amount of transport involved. In this study, an alternative approach to recycling has been tested using efficient small-scale biofuel boilers for co-combustion of biofuel and high-energy waste. The dry combustible fraction of source-sorted household waste was mixed with the energy crop reed canary-grass (Phalaris Arundinacea L.), and combusted in both a 5-kW pilot scale reactor and a biofuel boiler with 140 180 kW output capacity, in the form of pellets and briquettes, respectively. The chlorine content of the waste fraction was 0.2%, most of which originated from plastics. The HCl emissions exceeded levels stipulated in new EU-directives, but levels of equal magnitude were also generated from combustion of the pure biofuel. Addition of waste to the biofuel did not give any apparent increase in emissions of organic compounds. Dioxin levels were close to stipulated limits. With further refinement of combustion equipment, small-scale co-combustion systems have the potential to comply with emission regulations. PMID- 15823747 TI - From dumping to sanitary landfills--solid waste management in Israel. AB - To address the problem of solid waste in Israel, the Ministry of the Environment has formulated a policy based on integrated waste management. The policy calls for reduction of waste at source, reuse, recycling (including composting), waste to-energy technologies, and landfilling. Due to the implementation of this policy, all the large dumps were closed, state-of-the art landfills were built, and recovery rates have increased from 3% in the beginning of the 1990s to almost 20% in 2003. More than 95% of the municipal solid waste is disposed and treated in an environmentally sound manner - in comparison to a mere 10% just a decade ago. The policy was implemented utilizing both enforcement and financial support ("stick and carrot" approach). PMID- 15823748 TI - An estimation of the social costs of landfill siting using a choice experiment. PMID- 15823749 TI - An estimation of the social costs of landfill siting using a choice experiment. AB - The paper by Sasao on using a choice experiment (CE) to incorporate social costs in siting a landfill is a good step forward in siting landfills and other waste handling facilities in a more socially responsible manner. However, Sasao's method, which is well-established in the economics literature, has a number of underlying assumptions that practitioners must be careful to consider in applying the approach. The focus of this discussion is on three issues: (1) the form of utility function used, (2) the perceptions of the survey questions by the respondents versus the intended perception, and (3) the application of the results to situations beyond the assessed situations. This discussion article urges practitioners to carefully check their implementation of a choice experiment method to make sure the elicited values correctly represent those of the community. This discussion should not be construed as suggesting that the approach or results of Sasao (2004) are incorrect. Instead this discussion is meant to elaborate on the ideas and approach presented by Sasao with the hope that the approach will be applied more widely. PMID- 15823750 TI - BRCA2: breaks, mistakes and failed separations. AB - BRCA2 was identified in 1995, one year after BRCA1. In terms of knowledge of the function of its product, BRCA2 has remained the less well-characterised gene. Both BRCA1 and BRCA2 are closely implicated in the repair of double-strand breaks in DNA by homologous recombination, but beyond that a function for BRCA2 has been hard to discern. A recent study has extended the function of BRCA2 to the regulation of cell cleavage and separation. Other groups have also shown how BRCA2, RAD51 and DSS1 co-exist in a menage a trois and how the disruption of any one of the three cohabitants can have disastrous consequences for the cell. PMID- 15823751 TI - The role of eosinophils in the pathogenesis of asthma. AB - Two recent papers have addressed the ever-intriguing question of the role of eosinophils in asthma. Both groups used experimental models of airway inflammation in mice that were gene targeted to selectively ablate the eosinophil lineage. One group found that eosinophils were required for both airway hyperresponsiveness and mucus accumulation. The other demonstrated a 'critical role' for the cell in airway remodelling. The results, although largely confirmatory of previous studies both in mice and humans, put the eosinophil firmly back on the asthma stage and strengthen the case for developing effective eosinophil-depleting agents for clinical use. PMID- 15823752 TI - Uncovering the mysteries of invasive streptococcal diseases. AB - Group A streptococci, although considered extracellular pathogens, are capable of causing severe life-threatening invasive infections, such as necrotising fasciitis, bacteraemia and toxic-shock-like syndrome. A complete understanding of the mechanism by which these organisms cause invasive disease has been hampered by the extreme human specificity and high genetic diversity among group A streptococci. Three recent papers have uncovered some of the mysteries of streptococcal invasive diseases. Using two different technologies, these papers have contributed enormously towards our understanding of the molecular events underlying streptococcal invasive diseases. PMID- 15823753 TI - The controversy, potential and roles of spermatozoal RNA. AB - The majority of cellular and molecular andrologists endorse the view that the sperm is a vessel for transporting the paternal genome to the waiting egg and nothing more. Any requirement for additional spermatozoal components that enter the ooplasm apart from the paternal centriole and the soluble egg-activating factor is generally dismissed. Many studies, however, have reported RNAs in ejaculate spermatozoa and we now know that mRNAs are delivered to the egg on fertilisation. The function and utility of sperm mRNA remains essentially unexplored. Here, we examine the controversy surrounding spermatozoal mRNA carriage, the evidence refuting its presence as an artefact and how spermatozoal mRNA is leading us to suspect that, quite apart from its undoubted diagnostic potential, it might have an important role in the establishment and maintenance of a viable paternal genome. PMID- 15823754 TI - Tau phosphorylation in Alzheimer's disease: pathogen or protector? AB - During the past decade, hypotheses concerning the pathogenesis of most neurodegenerative diseases have been dominated by the notion that the aggregation of specific proteins and subsequent formation of cytoplasmic and extracellular lesions represent a harbinger of neuronal dysfunction and death. As such, in Alzheimer's disease, phosphorylated tau protein, the major component of neurofibrillary tangles, is considered a central mediator of disease pathogenesis. We challenge this classic notion by proposing that tau phosphorylation represents a compensatory response mounted by neurons against oxidative stress and serves a protective function. This novel concept, which can also be applied to protein aggregates in other neurodegenerative diseases, opens a new window of knowledge with broad implications for both the understanding of mechanisms underlying disease pathophysiology and the design of new therapeutic strategies. PMID- 15823755 TI - Alzheimer's disease: Abeta, tau and synaptic dysfunction. AB - Alzheimer's disease is a progressive neurodegenerative disorder that is characterized by two hallmark lesions: extracellular amyloid plaques and neurofibrillary tangles. The role that these lesions have in the pathogenesis of AD has proven difficult to unravel, in part because of unanticipated challenges of reproducing both pathologic hallmarks in transgenic mice. Recent advances in recapitulating both plaques and tangles in the brains of transgenic mice are leading to novel insights into their role in the degenerative process, including their impact on synaptic activity and plasticity. Transgenic mice that harbor both neuropathological lesions are also facilitating the elucidation of the relationship of these proteinaceous aggregates to one another and providing a crucial in vivo system for developing and evaluating therapies. PMID- 15823756 TI - Mechanisms of cell death in rhodopsin retinitis pigmentosa: implications for therapy. AB - Retinitis pigmentosa (RP) is a group of retinal degenerative diseases that are characterised primarily by the loss of rod photoreceptor cells. Mutations in rhodopsin are the most common cause of autosomal-dominant RP (ADRP). Here, we propose a new classification for rhodopsin mutations based on their biochemical and cellular properties. Several different potential gain-of-function mechanisms for rhodopsin ADRP are described and discussed. Possible dominant-negative mechanisms, which affect the processing, translocation or degradation of wild type rhodopsin, are also considered. Understanding the molecular and cellular consequences of rod-opsin mutations and the underlying disease mechanisms in ADRP are essential to develop future therapies for this class of retinal dystrophies. PMID- 15823757 TI - Recent advances in the genetic investigation of osteoarthritis. AB - Osteoarthritis (OA) demonstrates considerable clinical heterogeneity, generating heated debate over whether OA is a single disease or a complex mix of disparate diseases and concerning which tissues are principally involved in disease initiation and progression. Epidemiological studies have demonstrated a major genetic component to OA risk. However, these studies have also revealed differences in risk between males and females and for disease at different skeletal sites. This observation has resulted in the concept of genes for specific sites rather than a generalised OA phenotype. Recent breakthroughs have shed considerable light on the nature of OA genetic susceptibility. Many candidate genes have been confirmed, such as the interleukin-1 gene cluster and the oestrogen alpha-receptor gene ESR1. Genome-wide linkage scans have revealed several regions harbouring novel loci, some of which are beginning to yield their genes. PMID- 15823758 TI - Prohibitin: a potential target for new therapeutics. AB - Prohibitin (PHB) is localized to the mitochondria where it might have a role in the maintenance of mitochondrial function and protection against senescence. There is considerable controversy concerning the function of nuclear-localized PHB. PHB has potential roles as a tumor suppressor, an anti-proliferative protein, a regulator of cell-cycle progression and in apoptosis. PHB might also function as a cell-surface receptor for an as-yet unidentified ligand. Cell associated PHB in the gastrointestinal tract has been implicated in protection against infection and inflammation and the induction of apoptosis in other tissues. The diverse array of functions of PHB, together with the emerging evidence that its function can be modulated specifically in certain tissues, suggest that targeting PHB would be a useful therapeutic approach for the treatment of variety of disease states, including inflammation, obesity and cancer. PMID- 15823759 TI - Metabolic risk during antipsychotic treatment. AB - BACKGROUND: Compared with the general population, individuals with schizophrenia demonstrate an increased prevalence of obesity, type 2 diabetes mellitus (T2DM), and cardiovascular disease (CVD). Increased adiposity is associated with decreases in insulin sensitivity, leading to an increased risk of hyperglycemia and hyperlipidemia. Antipsychotic medications can increase adiposity, and a range of evidence from case reports, observational studies, retrospective database analyses, and controlled experimental studies (including randomized clinical trials) suggests that treatment with antipsychotic medications may be associated with an increased risk for insulin resistance, hyperglycemia, dyslipidemia, and T2DM. OBJECTIVE: This article reviews current evidence for the hypothesis that treatment with antipsychotic medications may be associated with increased risks for weight gain, insulin resistance, hyperglycemia, dyslipidemia, and T2DM, and examines the relationship of adiposity to medical risk. METHODS: Relevant publications were identified through a search of MEDLINE from 1975 to the present using the primary search parameters "diabetes or hyperglycemia or glucose or insulin or lipids" and "antipsychotic." Meeting abstracts and earlier nonindexed articles concerning antipsychotic-associated weight gain and metabolic disturbance were also reviewed. Key studies in this emerging literature were summarized, including case reports, observational studies, retrospective database analyses, and controlled experimental studies. RESULTS: Individual antipsychotic medications are associated with different degrees of treatment-induced increases in body weight and adiposity, ranging from modest effects (<2 kg) with amisulpride, ziprasidone, and aripiprazole to clinically significant increases with olanzapine (4-10 kg). In addition to strong evidence concerning the effect of adiposity on insulin sensitivity in nonpsychiatric populations, increased adiposity in patients with schizophrenia has been associated with decreases in insulin sensitivity; this and other effects may contribute to increases in plasma glucose concentrations and lipid levels. CONCLUSION: Metabolic changes in psychiatric patients who receive antipsychotic agents can contribute to the development of the metabolic syndrome and increase the risk for T2DM and CVD. PMID- 15823761 TI - Anakinra: review of recombinant human interleukin-I receptor antagonist in the treatment of rheumatoid arthritis. AB - BACKGROUND: Interleukin-1 (IL-1) plays an important role in the pathophysiology and progression of rheumatoid arthritis (RA) by contributing to destruction of cartilage, bone, and periarticular tissues. Inhibiting IL-1 synthesis or activity with the use of recombinant human IL-1 receptor antagonist (anakinra) may prove to be an effective approach to the treatment of RA. OBJECTIVE: The purpose of this article is to review the effects of anakinra in the treatment of RA. METHODS: A MEDLINE search from 1982 to 2003 was used to identify animal studies and randomized clinical trials of anakinra and other therapies that target IL-1. RESULTS: Clinical trials of anakinra have shown that it reduces the signs and symptoms of active disease and slows the rate of radiographic destruction in adults with RA. With anakinra 150 mg/d, 43% of patients achieved an American College of Rheumatology (ACR) 20% response, compared with 27% with placebo (P = 0.014). The ACR20 score indicates at least 20% improvement in the ACR composite score, which includes assessment of tender and swollen joint count, and other clinical end points such as pain and disability assessment. Patients treated with anakinra also experienced a 59% reduction in new bony erosion compared with controls (P < 0.001) and a 65% reduction in joint space narrowing as measured by the modified Sharp score (P = 0.020). Injection-site reactions were the most commonly reported adverse event, occurring in 50%, 73%, and 81% of patients receiving anakinra 30, 75, and 150 mg/d, respectively, compared with 25% of patients receiving placebo. Few serious adverse events were reported, and they typically occurred in patients receiving the highest daily dosage. CONCLUSIONS: IL-1 is an important cytokine in promoting the damage associated with RA. Anakinra is mildly to moderately effective and well tolerated in patients with active RA when used as monotherapy or in combination with methotrexate. PMID- 15823760 TI - Review of ibandronate in the treatment of metastatic bone disease: experience from phase III trials. AB - BACKGROUND: Metastatic bone disease is a serious clinical problem in patients with advanced cancer. Bisphosphonates inhibit the activity of osteoclasts and are the treatment of choice for bone metastases. OBJECTIVE: This article reviews the efficacy and safety data from Phase III trials of ibandronate in metastatic bone disease. METHODS: Phase III data (available as of June 2004) for ibandronate were reviewed. Literature searches using the MEDLINE database and abstracts from scientific meetings were used to obtain data from Phase III trials of ibandronate. RESULTS: Compared with placebo, patients with bone metastases from breast cancer receiving IV ibandronate (6 mg infused over 1-2 hours every 3-4 weeks) or oral ibandronate (50 mg/d for up to 96 weeks) had a statistically significant reduction in skeletal complications, as measured by the Skeletal Morbidity Period Rate (P = 0.004 vs placebo). Multivariate Poisson regression analysis of the data showed that the risk of a new bone event was reduced by 40% with IV ibandronate 6 mg and by 38% with oral ibandronate 50 mg, compared with placebo (P < or = 0.003). Both formulations also reduced bone pain below baseline levels over 2 years (P < or = 0.001 vs placebo). IV and oral ibandronate were well tolerated, with adverse-event profiles comparable to placebo and no significant renal toxicity. CONCLUSIONS: IV and oral ibandronate provide meaningful clinical benefits in patients with bone metastases from breast cancer. Both formulations reduce the risk of skeletal events and provide sustained relief from metastatic bone pain. With its favorable efficacy and safety profile, and the added convenience of the oral formulation, ibandronate provides improved treatment options for managing metastatic bone disease. PMID- 15823762 TI - Secondary hyperparathyroidism: review of the disease and its treatment. AB - BACKGROUND: Most patients with chronic kidney disease (CKD) stage 5 develop secondary hyperparathyroidism (SHPT). SHPT is an adaptive response to CKD and its associated disruptions in the homeostatic control of serum phosphorus, calcium, and vitamin D. The poor control of mineral and parathyroid hormone (PTH) levels characteristic of SHPT is associated with serious clinical consequences. OBJECTIVE: This review discusses the pathophysiology and consequences of SHPT, as well as the efficacy and limitations of current treatment modalities. METHODS: Literature searches were conducted using the MEDLINE, EMBASE, and BIOSIS databases. Additional information was obtained from Internet web sites, textbooks, and nephrology congress abstracts. RESULTS: Patients with uncontrolled SHPT are at higher risk for cardiovascular morbidity and mortality, hospitalization, bone disease, vascular and soft-tissue calcification, and vascular access failure than patients whose mineral and PTH levels are well managed. New National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) targets for calcium, phosphorus, calcium-phosphorus product, and PTH control have recently been published with the aim of improving the management of mineral metabolism in CKD patients. Data from observational studies suggest that the majority of patients currently have PTH and mineral levels outside these target ranges. CONCLUSIONS: Given the inadequacies of current therapies, novel agents are being developed that may help improve the management of SHPT. PMID- 15823763 TI - Clinical review of a long-acting, injectable formulation of risperidone. AB - BACKGROUND: A long-acting, injectable risperidone formulation is the first depot atypical antipsychotic drug to become available in the United States. OBJECTIVE: The intent of this article is to review the efficacy and safety data available for long-acting, injectable risperidone. METHODS: Information was identified via MEDLINE (years, 1990-May 2004) using the terms risperidone, long-acting injectable, depot, and delayed-action preparations. The manufacturer also provided information about risperidone in the form of abstracts and summaries of professional meetings. RESULTS: Several 12-week studies and one 12-month study suggest that long-acting risperidone is an effective and well-tolerated treatment option for the maintenance therapy of schizophrenia. Thus far, no unexpected adverse events have been reported with the long-acting formulation. Extrapyramidal symptoms with long-acting risperidone were uncommon, dose-related, and similar to those observed with oral risperidone in short-term trials. A small, dose related weight gain occurred with long acting risperidone, again similar to that seen with oral risperidone. Pain at the injection site was uncommon and decreased with continued administration. The long-acting, injectable formulation comes in an aqueous suspension of microspheres. There is no initial drug release after injection; the main release of risperidone begins at week 2 to 3 postinjection, increases during weeks 3 and 4, is maintained during weeks 4 through 6, and declines between weeks 6 and 7. With repeated injections every 2 weeks, steady-state levels are usually reached by weeks 6 to 8. For most patients, the initial dosage should be 25 mg every 2 weeks, and oral administration should continue for the first 3 weeks after initial injection. Doses can be increased every 8 weeks to a maximum of 50 mg every 2 weeks. CONCLUSION: Long-acting risperidone offers clinicians a combination of the benefits of a depot antipsychotic drug with the therapeutic advantages of an atypical antipsychotic drug. PMID- 15823764 TI - Combination oxycodone 5 mg/ibuprofen 400 mg for the treatment of postoperative pain: a double-blind, placebo- and active-controlled parallel-group study. AB - OBJECTIVE: This study compared the efficacy and safety of a single dose of oxycodone 5 mg/ibuprofen 400 mg versus its individual components and placebo in a third-molar extraction model. METHODS: In this multicenter, double-blind, double dummy, parallel-group investigation, subjects with moderate to severe pain within 5 hours after extraction of > or =2 ipsilateral bony impacted third molars were randomized to single doses of oxycodone 5 mg/ibuprofen 400 mg, ibuprofen 400 mg, oxycodone 5 mg, or placebo. Primary efficacy variables were the sum of pain intensity difference over 6 hours (SP1D6) and total pain relief through 6 hours (TOTPAR6). The pharmacokinetics of oxycodone and ibuprofen, alone and in combination, were also determined in a subset of patients. RESULTS: A total of 498 subjects were randomized to treatment (187 to oxycodone 5 mg/ibuprofen 400 mg, 186 to ibuprofen 400 mg, 63 to oxycodone 5 mg, and 62 to placebo). Baseline demographics were generally similar among treatment groups, despite differences in sex (P = 0.041) and race (P = 0.023). Combination therapy was associated with greater analgesia than ibuprofen alone, oxycodone alone, or placebo (mean [SE] TOTPAR6: 13.3 [0.52], 12.2 [0.52], 4.3 [0.82], and 4.2 [0.83], respectively [P < 0.001 vs oxycodone or placebo, P = 0.012 vs ibuprofen]; mean [SE] SP1D6: 6.54 [0.42], 5.41 [0.44], 0.14 [0.60], and 0.32 [0.59], respectively [P < 0.001 vs oxycodone or placebo, P = 0.002 vs ibuprofen]). Combination therapy was well tolerated. Pharmacokinetic results implied no interaction between oxycodone and ibuprofen. CONCLUSIONS: In this study, a single dose of oxycodone 5 mg/ibuprofen 400 mg was fast-acting, effective, and well tolerated in subjects with moderate to severe pain after dental surgery. Oxycodone 5 mg alone did not provide an efficacy benefit over placebo in this study. PMID- 15823765 TI - Pharmacokinetic properties of combination oxycodone plus racemic ibuprofen: two randomized, open-label, crossover studies in healthy adult volunteers. AB - BACKGROUND: As part of ongoing studies to evaluate the analgesic efficacy and pharmacokinetic properties of combination oxycodone plus ibuprofen in the treatment of moderate to severe acute pain, 2 pharmacokinetic studies were conducted. OBJECTIVES: The goals of these studies were to compare the pharmacokinetic properties of monotherapy with oxycodone or ibuprofen with those of a tablet formulation of these 2 agents combined (study A), and to determine whether the absorption of the individual agents when given in the combination tablet was affected by the concomitant ingestion of food (study B). METHODS: Study A was a single-center, open-label, randomized, single-dose, 3-period, 3 way, crossover study. Healthy male subjects received oxycodone 5 mg, ibuprofen 400 mg, or a combination tablet containing both, after an overnight fast of > or =8 hours, on study days 1, 8, and 15. Study B was a single-center, open-label, randomized, single-dose, single-crossover study. Healthy volunteers received a tablet containing a combination of oxycodone 5 mg plus ibuprofen 400 mg after either an overnight fast of > or =8 hours or a standardized high-fat breakfast. Both studies included a washout period of > or =7 days between treatments. In both studies, the pharmacokinetic properties (C(max), T(max), t(1/2), AUC(0-4), AUC(0-1), and AUC(0-infinity)) of oxycodone and ibuprofen were derived from plasma drug concentrations. Analysis of variance was used to determine and compare pharmacokinetic properties. RESULTS: Twenty-four healthy, white, male subjects were included in study A (mean age, 26.0 years; mean body weight, 71.3 kg; mean height, 170.0 cm). Study B involved 12 subjects (11 men, 1 woman; mean age, 24.8 years; mean body weight, 77.2 kg; mean height, 181.4 cm). The pharmacokinetic properties of ibuprofen and oxycodone were not statistically different when administered alone or combined. Food intake did not affect the rate of oxycodone absorption (90% Cl of C(max) of fasted state vs fed state, 103 130), or the rate (90% Cl of C(max) of fasted state vs fed state, 72-95) or extent (90% Cl of AUC(0-infinity) of fasted state vs fed state, 88-102) of ibuprofen absorption. The extent of oxycodone absorption was slightly increased when the combination was given with food (90% Cl of AUC(0-infinity) of fasted state vs fed state, 115-127). CONCLUSIONS: The single-dose pharmacokinetic profiles of oxycodone and ibuprofen in these healthy volunteers were similar when these 2 drugs were given as monotherapy or in combination, suggesting bioequivalence. Food intake before administration of a single dose of the combination did not affect ibuprofen absorption but marginally increased the extent, but not the rate, of oxycodone absorption. PMID- 15823766 TI - Cefdinir versus levofloxacin in patients with acute rhinosinusitis of presumed bacterial etiology: a multicenter, randomized, double-blind study. AB - BACKGROUND: Treatment guidelines for acute bacterial rhinosinusitis (ABRS) recommend 10 to 14 days of therapy with high-dose amoxicillin, amoxicillin/clavulanate, cefdinir, cefpodoxime, cefuroxime, a macrolide, or a newer fluoroquinolone, among other agents. OBJECTIVE: This study compared the clinical efficacy and tolerability of cefdinir and levofloxacin in patients with a diagnosis of acute rhinosinusitis of presumed bacterial origin. METHODS: In this multicenter, double-blind, noninferiority study, ambulatory adult patients who had signs and symptoms for >7 to 21 days before the screening visit and radiographic findings consistent with acute rhinosinusitis were randomized to receive cefdinir 600 mg or levofloxacin 500 mg, each once daily for 10 days. Clinical and radiologic response rates were determined at the test-of-cure (TOC) visit, which took place 9 to 14 days after the completion of treatment. RESULTS: Two hundred seventy-one patients (138 cefdinir, 133 levofloxacin) were enrolled and randomized to treatment at 27 study centers in the United States and Poland between November 2003 and March 2004. Of these, 241 (123 cefdinir, 118 levofloxacin) were clinically evaluable. The cefdinir group consisted of 75 women and 48 men, of whom 117 were white and 6 black; their mean (SD) age was 42.5 (14.3) years. The levofloxacin group consisted of 71 women and 47 men, of whom 111 were white and 7 black; their mean age was 40.4 (13.6) years. The 2 groups were similar in terms of presenting signs and symptoms and baseline radiographic findings. The most common presenting symptoms were sinus pain, sinus pressure, and purulent nasal discharge, each of which was reported by > or =89% of patients. Clinical cure rates at the TOC visit in the cefdinir and levofloxacin groups were 83% (102/123) and 86% (101/118), respectively (95% Cl for the difference in cure rates, -12.3 to 7.0). Cefdinir and levofloxacin were comparable in the treatment of infections classified as moderate to severe. The incidence of drug-related adverse events was generally comparable in the 2 treatment groups, although there were significant differences between cefdinir and levofloxacin in the incidence of vaginal moniliasis in women (11% vs 0%, respectively; P = 0.003), drug-related diarrhea (8% vs 1%; P = 0.005), and insomnia (0% vs 4%; P = 0.027). Only 2% of patients discontinued therapy prematurely as a result of a drug-related adverse event. CONCLUSION: In this population of patients with ABRS, the extended-spectrum cephalosporin cefdinir was as efficacious as the fluoroquinolone levofloxacin, suggesting that cefdinir may be a suitable alternative to the currently recommended fluoroquinolones. PMID- 15823767 TI - Combined therapy with insulin lispro Mix 75/25 plus metformin or insulin glargine plus metformin: a 16-week, randomized, open-label, crossover study in patients with type 2 diabetes beginning insulin therapy. AB - OBJECTIVE: This study aimed to assess glycemic response to a mixture of 75% insulin lispro protamine suspension and 25% insulin lispro (Mix 75/25) BID plus metformin versus insulin glargine QD plus metformin in patients with type 2 diabetes mellitus (DM). METHODS: Adults new to insulin therapy were enrolled in a multicenter, randomized, prospective, open-label, crossover study with 16 weeks on each treatment. Variables included glycosylated hemoglobin (HbA(1c)), hypoglycemia rate, fasting blood glucose (FBG), 2-hour postprandial blood glucose (ppBG), and rise in blood glucose after meals. RESULTS: One hundred five patients (mean age, 55 years) were randomized. There was no difference in baseline mean values for either treatment sequence group for body mass index, duration of DM, or HbA(1c). Ninety-five patients completed the study and 67 were included in the efficacy analysis. Mix 75/25 was associated with lower mean (SD) HbA(1c) at end point (7.4% [1.1%] vs 7.8% [1.1%]; P = 0.002). More patients using Mix 75/25 achieved target HbA(1c) < or =7.0% (42% [30/71] vs 18% [13/71]; P < 0.001). With Mix 75/25, the mean (SD) 2-hour ppBG was similar after lunch but lower after breakfast (156.4 [43.6] vs 171.1 [44.9] mg/dL; P = 0.012) and dinner (164.8 [42.5] mg/dL vs 193.8 [51.0] mg/dL; P < 0.001), although FBG was higher (139.3 [36.6] mg/dL vs 123.9 [34.9] mg/dL; P < 0.001). Rise in ppBG was lower with Mix 75/25 after breakfast (16.9 [47.0] mg/dL vs 47.4 [34.8] mg/dL; P < 0.001) and dinner (14.2 [44.1] mg/dL vs 45.9 [41.3] mg/dL; P < 0.001). Gain in mean (SD) body weight was greater with Mix 75/25 than insulin glargine (2.3 [4.0] kg vs 1.6 [4.0] kg; P = 0.006). For all randomized patients, mean (SD) hypoglycemia rates were lower with insulin glargine (0.68 [1.38] vs 0.39 [1.24] episodes/patient per 30 days; P = 0.041), although nocturnal hypoglycemia was similar. CONCLUSION: In this study population, Mix 75/25 plus metformin was associated with lower HbA(1c) than insulin glargine plus metformin, smaller rise in ppBG after breakfast and dinner, and higher proportion of patients achieving HbA(1c) < or =7.0%, with a slight increase in overall (but not nocturnal) hypoglycemia. PMID- 15823768 TI - Effects of escin on indinavir crystallization time in the urine of patients with HIV-I infection: a multicenter, randomized, open-label, controlled, four-period crossover trial. AB - BACKGROUND: The combination of indinavir, a protease inhibitor, and reverse transcriptase inhibitors is widely used in the treatment of HIV-1 infection. However, precipitation of indinavir crystals in the renal tubular lumen due to the drug's aqueous insolubility may result in characteristic symptoms of flank pain or classic renal colic. An in vitro study has shown that addition of escin to synthetic urine containing indinavir delayed the crystallization time of indinavir. OBJECTIVE: This study examined the efficacy and tolerability of the addition of escin to highly active antiretroviral therapy containing indinavir to delay the crystallization time of indinavir in urine. METHODS: This was a multicenter, randomized, open-label, controlled, 4-period crossover trial in which each period lasted 4 weeks. HIV-1-infected adults receiving treatment with indinavir plus 2 nucleoside analogue reverse-transcriptase inhibitors in whom plasma viral loads had been undetectable (HIV-1 RNA <200 copies/mL) for at least 6 months were randomly assigned to 1 of 2 groups based on the timing of the initiation of escin. Group I received escin during the second and third treatment periods, and group II received escin during the first and fourth treatment periods. The primary end point was the in vitro crystallization time of indinavir in 24-hour urine specimens, determined at the end of each 4-week period. Tolerability was assessed based on the number of patients with a rebound in plasma viral load and on the numbers of clinically and biologically relevant adverse events (including those requiring discontinuation of treatment). Clinical and laboratory evaluations were performed throughout each 4-week period. RESULTS: Fifty HIV-1-infected patients were enrolled, 47 were randomized to treatment (40 [85.1%] men, 7 [14.9%] women; median [interquartile range] age, 36 [34-45] years), and 30 completed the study. Urine pH and plasma and urine indinavir concentrations were unaffected by the addition of escin to antiretroviral treatment. The mean time to the onset of crystallization was 14.7 minutes with escin (95% Cl, 11.8-17.5) and 9.9 minutes without it (95% Cl, 6.7-13.1). Therefore, the addition of escin increased the mean crystallization time by 5.5 minutes (95% Cl, 1.5-9.5; P = 0.008), representing the overall capacity of study treatment to inhibit indinavir crystallization in the urine. Three of 47 patients had mild gastrointestinal symptoms associated with escin treatment. No episodes of nephrolithiasis were recorded during the study or after the completion of study treatment. CONCLUSION: The results of this prospective clinical trial of the effect of escin on indinavir crystallization time support the possibility that indinavir-associated nephrolithiasis may be prevented by means other than overhydration. Further research is needed in greater numbers of patients over longer follow-up times. PMID- 15823769 TI - Effect of zonisamide on the pharmacokinetics and pharmacodynamics of a combination ethinyl estradiol-norethindrone oral contraceptive in healthy women. AB - BACKGROUND: Several antiepileptic drugs have clinically significant pharmacokinetic interactions with oral contraceptives (OCs) that may result in contraceptive failure. OBJECTIVE: The aim of this study was to assess the effect of zonisamide on the pharmacokinetics of the individual components of a combination OC (ethinyl estradiol [EE] 0.035 mg and norethindrone [NOR] 1 mg) and on pharmacodynamic variables that may be increased in the event of reduced contraceptive efficacy (concentrations of serum luteinizing hormone [LH], follicle-stimulating hormone [FSH], and progesterone). METHODS: This was a single center, open-label, 1-sequence, crossover study. Healthy, premenopausal women received the combination OC for three 28-day cycles (combination OC for 21 days, followed by placebo for 7 days). Following stabilization on the OC during the first cycle, blood was collected during cycle 2 for the determination of serum EE and NOR profiles (day 14) and serum LH, FSH, and progesterone concentrations (days 13-15). Starting on day 15 of cycle 2, zonisamide was administered orally at 100 mg/d and titrated to a target dose of 400 mg/d. EE and NOR profiles and serum LH, FSH, and progesterone concentrations were obtained again in cycle 3 (in the presence of zonisamide) and compared with those from cycle 2 (in the absence of zonisamide). RESULTS: Thirty-seven healthy premenopausal women (mean age, 26.1 years [range, 18-51 years]; mean body weight, 65.5 kg [range, 50.4-93.1 kg]; mean height, 165.8 cm [range, 152.4-182.9 cm]) received > or =1 dose of zonisamide. Of the 33 subjects (89.2%) who completed the study, 26 (78.8%) underwent titration to a stable zonisamide dose of 400 mg/d. For EE, the mean (SD) AUC over a 24-hour dosing interval (AUC(tau)) was 1139 (317) pg.h/mL in cycle 2 and 1143 (312) pg.h/mL in cycle 3; the mean C(max) in the respective cycles was 133 (39) and 141 (46) pg/mL. For NOR, the corresponding values were 140 (48) and 159 (46) ng.h/mL for AUC(tau) and 21 (5.4) and 23 (6.7) ng/mL for C(max). The 90% Cls for the geometric mean ratios (cycle 3:cycle 2) for AUC(tau) and C(max) fell within the accepted range for lack of interaction (0.80-1.25). There were no increases in LH, FSH, or progesterone concentrations between cycle 2 and cycle 3. CONCLUSIONS: In these healthy volunteers, steady-state zonisamide dosing had no clinically significant effect on the pharmacokinetics of EE or NOR. There was no pharmacodynamic evidence that zonisamide is likely to reduce the contraceptive effectiveness of OCs containing EE and NOR. PMID- 15823770 TI - Adherence to a fixed-dose combination of rosiglitazone maleate/metformin hydrochloride in subjects with type 2 diabetes mellitus: a retrospective database analysis. AB - BACKGROUND: In 2002, fixed-dose combination therapy (FDCT) with rosiglitazone maleate plus metformin hydrochloride became available for the treatment of type 2 diabetes mellitus (DM-2) in subjects whose disease was uncontrolled on monotherapy with metformin or a thiazolidinedione. FDCT allows a reduced pill burden and a less complex medication regimen. OBJECTIVE: The objective of this study was to assess changes in medication adherence rates associated with oral hypoglycemic agents in subjects switching from either monotherapy or dual therapy with metformin and/or rosiglitazone to rosiglitazone-metformin FDCT. METHODS: In this retrospective database analysis, data were obtained from the pharmacy claims database of a large health benefits company. Prescription claims for subjects aged > or =18 years with DM-2 whose disease was uncontrolled on monotherapy with metformin or a thiazolidinedione were analyzed over a 12-month study period (a 6 month preindex period and a 6-month postindex period). Some subjects were receiving monotherapy with either metformin or rosiglitazone during the preindex period and remained on monotherapy throughout the postindex period (Mono/Mono cohort), switched to dual therapy with both agents (Mono/Dual cohort), or switched to FDCT (Mono/FDCT cohort). Some subjects were receiving dual therapy with metformin and rosiglitazone during the preindex period and remained on dual therapy throughout the postindex period (Dual/Dual cohort) or switched to FDCT (Dual/FDCT cohort). A medication possession ratio (MPR)-a proxy measurement of medication adherence-was calculated for each subject for each period. Changes in medication adherence were compared using a general linear model. RESULTS: Overall, data from the records of 16,928 subjects (8499 men, 8429 women; mean [SD] age, 58.12 [11.97] years) were included in this study. There was significantly less reduction in the MPR change for the Mono/FDCT cohort compared with the Mono/Dual cohort (-4.6% vs -12.4%; P < 0.001). There was significant improvement in the mean MPR change for the Dual/FDCT cohort compared with the Dual/Dual cohort (3.5% vs -1.3%; P < 0.005). CONCLUSIONS: The results of this retrospective database analysis suggest that rosiglitazone-metformin FDCT yielded significant improvements in medication adherence rates compared with dual therapy regimens. PMID- 15823771 TI - Comparison of the pharmacokinetics of lansoprazole 15- and 30-mg sachets for suspension versus intact capsules. AB - OBJECTIVE: The pharmacokinetic profiles of single doses of lansoprazole 15- and 30-mg sachets for suspension were compared with those of corresponding doses of lansoprazole oral capsules. METHODS: Healthy adult male and female subjects were randomized (1:1 ratio) into 2 Phase 1, open-label, single-dose, 2-sequence, 2 period complete crossover studies. In the first study, each subject received 1 lansoprazole 15-mg sachet mixed with water and 1 lansoprazole 15-mg oral capsule; in the second study, each subject received 1 lansoprazole 30-mg sachet mixed with water and 1 lansoprazole 30-mg oral capsule. Administration of the 2 formulations was separated by a washout period of > or =7 days. Blood samples were collected before and after each administration to assess the pharmacokinetic parameters of lansoprazole and bioequivalence between suspension and capsule. RESULTS: Thirty six subjects (19 males, 17 females) with a mean (SD) age of 32.0 (9.6) years and mean (SD) body weight of 68.6 (10.5) kg received lansoprazole 15 mg. Thirty-six subjects (22 males, 14 females) with a mean (SD) age of 38.0 (8.3) years and mean (SD) body weight of 75.1 (9.7) kg received lansoprazole 30 mg. The pharmacokinetic parameters of the 15- and 30-mg lansoprazole sachets for suspension were similar to those of the corresponding doses of the oral capsules. The mean (SD) values for C(max) and AUC from time 0 to infinity (AUC(0-infinity) for the lansoprazole 15-mg sachet (591.9 [242.3] ng/mL and 1614 [2065] ng.h/mL, respectively) did not differ significantly from those for the lansoprazole 15-mg capsules (578.6 [275.2] ng/mL and 1620 [2290] ng.h/mL, respectively). These parameters also did not differ significantly between the lansoprazole 30-mg sachet and 30-mg capsule: mean (SD) C(max), 1103 (428.3) and 1077 (465.6) ng/mL, respectively; mean (SD) AUC(0-infinity), 2655 (1338) and 2669 (1311) ng.h/mL, respectively. The 90% Cls for C(max) and AUC(0-infinity) ratios were contained within the 0.80 to 1.25 equivalence range, supporting bioequivalence. CONCLUSIONS: These findings suggest that the 15- and 30-mg lansoprazole sachets for suspension are bioequivalent to the corresponding doses of oral capsules. The sachet for suspension may provide an alternative route of administration to patients who have difficulty swallowing solid oral formulations. PMID- 15823772 TI - Comparison of pharmacokinetic and pharmacodynamic properties of single-dose oral insulin spray and subcutaneous insulin injection in healthy subjects using the euglycemic clamp technique. AB - BACKGROUND: Oral insulin spray is a new, noninjectable method of insulin delivery. This system delivers an aerosol of uniform-sized droplets containing regular human insulin at a high velocity into the oropharyngeal cavity for local transmucosal absorption. OBJECTIVE: The purpose of this study was to compare the pharmacokinetic and pharmacodynamic properties of single-dose oral insulin spray and SC insulin injection in healthy subjects. METHODS: Healthy male volunteers aged 21 to 25 years participated in this open-label study conducted at the Diabetes Unit, Hadassah-Hebrew University Hospital, Jerusalem, Israel. Subjects presented at 2 visits separated by 7 to 14 days. At both visits, the euglycemic clamp technique was used to maintain a constant blood glucose level. At one visit, subjects received regular human insulin 0.1 U/kg by SC injection. At the other visit, subjects received 15 puffs (150 U) of oral insulin spray. The pharmacokinetic (insulin absorption) and pharmacodynamic (glucose uptake) properties of the drugs were evaluated using blood analyses over the subsequent 360 minutes. RESULTS: Six volunteers were enrolled (mean [SD] age, 22.8 [1.2] years; mean [SD] body mass index, 23.2 [2.2] kg/m(2)). The mean (SD) baseline corrected C(max) was significantly higher with oral insulin spray compared with SC insulin (54.0 [20.3] vs 30.8 [6.1] microU/mL; P = 0.028). Mean (SD) T(max) was significantly shorter with oral insulin spray compared with SC insulin (23.3 [5.2] vs 83.3 [42.2] minutes; P = 0.027). The mean (SD) time to maximal metabolic effect (maximum glucose infusion rate [GIR(max)]) (44.2 [8.6] vs 100.0 [35.6] minutes) and late time to half-maximal effect (101.0 [41.0] vs 257.2 [27.8] minutes) were shorter with oral insulin spray compared with SC insulin (both, P = 0.028). The baseline-corrected GIR(max) (6.8 [3.3] vs 6.2 [2.3] mg/kg . min) and glucose consumption (396.7 [178.0] vs 432.1 [226.0] mg/kg) during the 120 minutes after study drug administration were comparable between oral and SC insulin, respectively. CONCLUSIONS: In this study in a small, selected population of healthy male subjects under euglycemic conditions, oral insulin spray was associated with a higher C(max), shorter T(max), and faster time to peak glucose uptake compared with SC insulin. The short T(max) and the 120-minute duration of effect of oral insulin spray suggest it may be a promising alternative for fulfilling meal-related insulin requirements in persons with diabetes. PMID- 15823773 TI - Relative bioavailability and pharmacokinetics of a new sibutramine formulation in healthy male subjects: a randomized, open-label, two-period, comparative crossover study. AB - BACKGROUND: Sibutramine is an orally administered, centrally acting antiobesity drug. Sibutramine hydrochloride monohydrate is the conventional formulation, whereas sibutramine mesylate hemihydrate is a newly developed formulation. Drugs formed from different salts may differ in their solubility profiles and dissolution rates, which may affect their rate of absorption and thus their onset, duration, and intensity of effect. OBJECTIVE: This study was conducted to compare the relative bioavailability and pharmacokinetics of the new sibutramine formulation (test) with those of the conventional formulation (reference). METHODS: : This was a single-center, randomized, open-label, 2-period, comparative crossover study in healthy male subjects. All subjects received a single 15-mg oral dose of sibutramine hydrochloride monohydrate (reference) and a single 17.3-mg oral dose of sibutramine mesylate hemihydrate (test), both containing 12.55 mg sibutramine base. The 2 doses were separated by a 2-week washout period. Blood samples for pharmacokinetic analysis were collected during a 72-hour period after treatment. Safety parameters were assessed, including adverse events, hematology and biochemistry, urinalysis, and electrocardiography. Plasma concentrations of the active metabolites of sibutramine (desmethylsibutramine [M1] and didesmethylsibutramine [M2]) were determined, and the pharmacokinetic characteristics of the 2 formulations were compared using noncompartmental analysis. RESULTS: Sixteen subjects (mean [SD] age, 24.3 [2.3] years [range, 20-25 years]; mean [SD] body weight, 66.1 [5.1] kg [range, 57-77 kg]) were enrolled in and completed the study. The plasma concentration-time profiles of M1 and M2 were similar after administration of both formulations. The reference and test formulations showed pharmacokinetic equivalence with respect to M1 and M2. The relative bioavailability of the test drug was 117.6% for M1 and 102.4% for M2. The 90% Cls for the ratios of the log-transformed C(max) and AUC values were within the predetermined equivalence range of 80% to 125%. There were no significant changes in physical, biochemical, hematologic, or urinalysis variables during the study. Neither formulation was associated with any serious adverse events. CONCLUSION: In this study in healthy male subjects, the 2 sibutramine formulations were pharmacokinetically equivalent, and the newly developed formulation had a safety profile comparable to that of the conventional formulation. PMID- 15823774 TI - Gold standard medical therapy for glaucoma: defining the criteria identifying measures for an evidence-based analysis. AB - BACKGROUND: Over the past decade, several new medical therapies have become available for the treatment of primary open-angle glaucoma (POAG). A systematic evidence-based approach for identifying an optimal therapeutic agent is lacking. OBJECTIVES: The aims of this review were to critically evaluate published treatment recommendations for POAG and, based on a systematic review of the literature, to develop criteria that would define a "gold standard" medical therapy that reflects new treatment advances and established therapeutic goals. METHODS: A MEDLINE search spanning the years 1966 to 2002 and using the search terms gold standard, drug of choice, agent of choice, benchmark, ophthalmology, eye, and glaucoma was conducted and the results reviewed by a panel of 15 experts in the field of glaucoma. Published treatment recommendations for POAG were discussed. Criteria, anchored to medical evidence, for distinguishing a standard of medical therapy for POAG were defined. RESULTS: The terms connoting a gold standard therapy were found in only 258 of approximately 368,000 ophthalmology related citations and 53 of almost 23,000 glaucoma citations, validating the need to define therapeutic standards. The lack of recommendations for the use of new classes of ocular hypotensive agents was acknowledged. Criteria identified to evaluate intraocular pressure (IOP)-lowering agents as gold standards included the following: efficacy in reducing IOP consistently over a 24-hour period to a level that will preserve the visual field and protect the optic nerve without inducing tachyphylaxis and tolerance, paucity of local and systemic adverse effects, promotion of patient compliance, and applicability in diverse patient populations. CONCLUSIONS: These criteria should be employed as measures for evidence-based analyses to evaluate available and future IOP-lowering medical therapies for POAG. The conceptual framework presented may be applicable to other therapeutic areas. PMID- 15823775 TI - Effects of travoprost 0.004% ophthalmic solution, six weeks after its laminated packaging had been removed, in primary open-angle glaucoma: a randomized, controlled, investigator-blinded study. AB - BACKGROUND: Primary open-angle glaucoma (POAG) is a chronic and progressive optic nerve and retinal nerve fiber layer neuropathy, with characteristic visual field damage. The intraocular pressure (IOP) is typically higher than the level considered statistically normal. Although there is no known cure, appropriate reduction of IOP with hypotensive drugs (eg, the topical prostaglandin analogue travoprost) delays the progression of POAG. Chemical-stability studies of travoprost performed by the manufacturer suggest that the stability of travoprost is maintained beyond the expiration date, which is 6 weeks after the laminated packaging has been opened. OBJECTIVE: The goal of this study was to assess the efficacy and tolerability of travoprost 0.004% ophthalmic solution, 6 to 12 weeks after its expiration date, in patients with POAG. METHODS: This randomized, controlled, investigator-blinded study was conducted at 2 centers in Brazil: the Ophthalmology Department, Federal University of Goias, Goiania, and the Ophthalmology Department, Santa Casa de Misericordia Hospital in Sao Jose do Rio Preto, Sao Paulo. Patients with POAG (in 1 or both eyes) were randomly assigned to receive travoprost, either from a bottle from which the laminated packaging had been removed and that had been stored at room light and temperature for 6 weeks (ie, after the expiration date; opened group), or from a bottle that had been sealed until first use by the patient (control group). Drug was to be administered, 1 drop in the lower conjunctival sac (in the affected eye[s]), QD between 7 pm and 9 pm, for 6 weeks. IOP was measured at study weeks 0 (baseline), 4, and 6. The 2 treatment groups were compared with regard to hypotensor effect and incidence of adverse events (AEs). RESULTS: : Thirty-one patients completed the study (55 eyes; 28 right and 27 left eyes; 35 eyes of women, 20 eyes of men). The mean (SD) ages of the opened and control groups were 61.8 (13.5) and 62.8 (14.1) years, respectively. Twenty-four patients were included in both treatment groups (ie, 1 eye per group). The baseline IOP was similar between the 2 treatment groups. There was a significant reduction in IOP in both groups at 4 and 6 weeks (both, P < 0.001 vs baseline). However, no significant differences in IOP were found between the 2 treatment groups at any time during the study. Conjunctive hyperemia and a burning sensation in the eye immediately after application were the only AEs reported; the incidence of these was similar between the 2 treatment groups. CONCLUSIONS: In this study of patients with POAG, IOPs and AEs were similar in eyes receiving 6 weeks of treatment with travoprost 0.004% ophthalmic solution, either from bottles from which the laminated packaging had been opened and that had been stored at room light and temperature for 6 weeks (ie, after the expiration date), or from bottles that had been sealed until first use by the patient. These results suggest that travoprost remains effective for at least 12 weeks after the laminated packaging has been opened. PMID- 15823776 TI - Administration of the first dose of palivizumab immunoprophylaxis against respiratory syncytial virus in infants before hospital discharge: what is the evidence for its benefit? AB - BACKGROUND: Palivizumab is 1 of 2 agents used to prevent severe lower respiratory tract disease due to respiratory syncytial virus (RSV) infection. The American Academy of Pediatrics and the American College of Obstetricians and Gynecologists recommend administering the first dose of RSV immunoprophylaxis to eligible infants before hospital discharge. Unfortunately, third-party payers frequently do not separately reimburse administration of this therapy to hospitalized infants. OBJECTIVE: Because palivizumab is commonly used to provide RSV immunoprophylaxis, we systematically reviewed all published data on this drug to determine whether the evidence supports the recommendation of administering the first dose before hospital discharge. METHODS: MEDLINE was searched for all articles published in English from January 1, 1996, to October 31, 2003, using the search terms palivizumab and Synagis, and the following data were extracted onto a standardized form: author(s), year of publication, study design, patient population, sample size, criteria used for administration of RSV prophylaxis, location of palivizumab prophylaxis (inpatient or outpatient), parental satisfaction with administration of prophylaxis, incidence of RSV infection, and hospitalization rates for RSV. All selected publications were reviewed to determine whether they reported differences in the incidence of RSV infection or hospitalization in patients who received palivizumab before discharge compared with those who received it after discharge. Only those publications that specifically documented administration of the first dose of palivizumab before hospital discharge were included in the final analysis. RESULTS: Six of the 166 studies reviewed met the selection criteria. Although all 6 studies reported reduced RSV hospitalization rates with palivizumab prophylaxis, no study directly compared inpatient and outpatient administration with regard to parental satisfaction or rates of RSV infection or hospitalization. Furthermore, based on the data in these studies, it was not possible to detect any differences in parental satisfaction or rates of RSV infection or hospitalization between the 2 locations of administration. CONCLUSIONS: Based on our literature review, there is no evidence to support the recommendation that palivizumab be administered before hospital discharge in every infant who meets the criteria for RSV immunoprophylaxis. Eligible infants may be given the initial dose of RSV prophylaxis as outpatients, reducing the cost to institutions that currently provide palivizumab before hospital discharge. PMID- 15823777 TI - Patient-reported outcomes: D j Vu? PMID- 15823778 TI - Asthma treatment needs: a comparison of patients' and health care professionals' perceptions. AB - BACKGROUND: Patients' perceptions of asthma tend to differ from those of clinicians, who primarily focus on asthma control. Patients' treatment needs and preferences may not be adequately addressed. OBJECTIVE: The aims of this study were as follows: to provide data on unmet treatment needs and to investigate the main finding of a qualitative study using a questionnaire study. METHODS: To assess treatment needs in patients with asthma, focus groups were conducted with patients/parents and clinicians. Based on these results, quantitative surveys of adult patients and parents were performed in the United Kingdom, Germany, and Spain. RESULTS: The UK focus group comprised 11 patients and 8 parents; in Germany, there were 10 patients and 11 parents; in Spain, there were 5 patients and 8 parents. The focus groups showed some differences between clinicians' and patients'/parents' perceptions of treatment. For patients, side effects meant long-term effects (ie, 10-20 years); for clinicians, it meant occasional local problems. The quantitative study comprised 454 participants: 310 adult patients (mean [SD] age, 37.13 [13.12] years) and 142 parents (children's mean [SD] age, 13.98 [1.37] years), plus 2 nonspecified. Some patients reported good asthma control and simultaneously reported frequent exacerbations. Most patients and parents expressed a preference for a simpler regimen using fewer drugs, and most had concerns about their treatment. Although some patients concurred with treatment guidelines, 62.2% tended to rely on reliever medication (ie, bronchodilators). Additionally, 6.9% described their asthma as very well controlled but reported experiencing asthma symptoms > or =3 days per week. Finally, 1.9% of patients and 2.1% of parents reporting very well-controlled asthma also reported visiting the emergency department or calling a physician for a home visit in the previous 3 months. CONCLUSIONS: Asthma patients and parents of asthmatic children had unmet treatment needs and may interpret medical terminology differently than clinicians. PMID- 15823779 TI - Patterns of health care resource utilization after macrolide treatment failure: results from a large, population-based cohort with acute sinusitis, acute bronchitis, and community-acquired pneumonia. AB - BACKGROUND: Macrolide antibiotics are used as first-line therapy for the treatment of respiratory tract infections. The recent emergence of macrolide resistant pathogens is a major concern. OBJECTIVE: This study quantifies the frequency of macrolide treatment failure in respiratory infections and examines its impact on health care use. METHODS: Patients with respiratory infections treated with macrolides in outpatient clinics from January to December 2002 were identified from a health insurance claims database. Macrolide treatment failure was defined as the receipt of a second antibiotic, different from the first, within 4 weeks after the initial macrolide. The end points were numbers of hospitalizations and emergency department and office visits within 1 month after the initial macrolide. We examined diagnostic codes on claim forms for posttreatment hospitalizations and visits to identify those most likely to be related to treatment failure as opposed to other causes. Utilization data were analyzed by Poisson regression to control for confounding variables. RESULTS: The patients were divided into acute sinusitis (n = 111,135), acute bronchitis (n = 157,360), and community-acquired pneumonia (n = 36,212). Of these respective groups, 11,285 (10.2%), 15,498 (9.9%), and 4144 (11.4%) received a second antibiotic within 4 weeks. This subgroup with macrolide treatment failure was older, included more women, and had used more medical care before the index visit compared with patients with treatment success. After adjustment for age, sex, and previous health care use, patients experiencing treatment failure were more likely to be admitted to the hospital or to use emergency department or outpatient care after the index visit. This association was strongest for admissions and visits pertaining to the care of respiratory infections. CONCLUSIONS: By our definition, about 10% of patients with respiratory infections who were treated with macrolide antibiotics experienced treatment failure within 4 weeks. Macrolide treatment failure was associated with increased health care utilization. PMID- 15823780 TI - From Don Giovanni to Magic Johnson: methodological conundrums in the measurement of sexual risk behavior. PMID- 15823781 TI - Commentary on Schroder et al. (2003a, 2003b). PMID- 15823783 TI - Perceptions of local HIV/AIDS prevalence and risks for HIV/AIDS and other sexually transmitted infections: preliminary study of intuitive epidemiology. AB - BACKGROUND: Risk perceptions are theoretically important in predicting health behavior but have only shown modest evidence in predicting sexual risk behavior. PURPOSE: Our purpose is to investigate whether perceptions of the local prevalence of disease serve as a predictor of health behavior, particularly behaviors associated with infectious diseases such as HIV/AIDS. METHODS: Four hundred eighty-seven men and 236 women receiving diagnostic and treatment services at a sexually transmitted infections (STI) clinic in a moderate-size U.S. city completed anonymous surveys of perceived prevalence of HIV/AIDS and other STI and sexual risk and protective behaviors. RESULTS: Participants who estimated a lower AIDS burden in their city relative to other U.S. cities demonstrated greater numbers of sex partners, higher rates of sexual risk practices, and higher rates of STI. They were also less likely to have been tested for HIV. CONCLUSIONS: This initial study of intuitive epidemiology suggests that STI clinic patients may have a sense for the relative burden of AIDS in their city and estimates of local disease prevalence may predict sexual risk behaviors. PMID- 15823784 TI - Interactive effects of avoidant coping and parental hypertension on rate pressure product reactivity in women. AB - BACKGROUND: Previous research suggests that personality, situational context variables, and genes might interact to potentiate cardiovascular stress responses. PURPOSE: Our purpose is to examine interactive effects of dispositional avoidant coping and parental hypertension on cardiovascular reactivity to three different laboratory stressors. METHOD: Participants were 63 healthy female students. Stressors were an evaluated videotaped speech, the cold pressor, and viewing of the speech video. Heart rate and blood pressure were continuously recorded during baselines and tasks. RESULTS: After controlling for age, body mass index, smoking status, reported exercise, alcohol consumption, oral contraceptive use, parental hypertension, and avoidant coping, the interaction term of parental hypertension and avoidant coping was the best predictor of rate pressure product (RPP) reactivity to the speech task and to the video-viewing task. In contrast, reactivity to the cold pressor was not significantly predicted by this interaction term. Furthermore, there were no significant main effects of parental hypertension and avoidant coping on RPP reactivity in any of the tasks. There were no significant interactive effects for diastolic blood pressure. CONCLUSIONS: Consistent with an interactive model of cardiovascular reactivity, parental hypertension plus avoidant coping led to enhanced RPP reactivity. This effect, however, was only evident during self threatening stressors. The results suggest that the coincidence of genetic and personality factors might enhance risk for cardiovascular diseases. PMID- 15823785 TI - The effects of hypnotic and nonhypnotic imaginative suggestion on pain. AB - BACKGROUND: Few studies have compared placebo and suggested pain reduction. PURPOSE: Hypnotic and nonhypnotic imaginative analgesia suggestions were compared against a placebo in reducing experimental pain. The mediator role of response expectancies and the moderator role of hypnotic and nonhypnotic imaginative suggestibility were evaluated. METHODS: Sixty participants previously assessed for hypnotic and nonhypnotic imaginative suggestibility were assigned to one of two experimental conditions or a no-treatment control condition. In the "placebo first" condition, participants received placebo, followed by imaginative and then hypnotic analgesia suggestions. In the "placebo last" condition, participants received imaginative and then hypnotic suggestions, followed by placebo. RESULTS: Imaginative and hypnotic suggestions did not differ significantly and were more effective than no treatment in reducing pain. The placebo was no different from the analgesia suggestions and was more effective than no treatment, but only when administered after the suggestions. Pain reduction was mediated by expectancy but was not significantly related to suggestibility or hypnotizability, the latter operationalized as hypnotic suggestibility with imaginative suggestibility statistically controlled. CONCLUSIONS: In the general population, nonhypnotic imaginative suggestions may be as effective as hypnotic suggestions in reducing pain. Response expectancies would seem to be an important mechanism of placebo and suggested pain reduction. PMID- 15823786 TI - Effects of the mediterranean lifestyle program on multiple risk behaviors and psychosocial outcomes among women at risk for heart disease. AB - BACKGROUND: The Mediterranean Lifestyle Program was evaluated for its effects on multiple behavioral risk factors for coronary heart disease (CHD) among postmenopausal women with diabetes. PURPOSE: Our purpose is to test a comprehensive lifestyle management intervention to reduce CHD risk in postmenopausal women with type 2 diabetes. METHODS: Participants (N = 279) were randomized to usual care (UC) or Mediterranean Lifestyle Program, a lifestyle change intervention aimed at the behavioral risk factors (eating patterns, physical activity, stress management, and social support) affecting risk for CHD in postmenopausal women with type 2 diabetes. RESULTS: In original and intent-to treat analyses, Mediterranean Lifestyle Program participants showed significantly greater improvement in dietary behaviors, physical activity, stress management, perceived support, and weight loss at 6 months compared to UC. CONCLUSIONS: This study demonstrated the effectiveness of the Mediterranean Lifestyle Program in improving self-care among women with type 2 diabetes, showed that postmenopausal women could make comprehensive lifestyle changes, and provided evidence that a program using social-cognitive strategies and peer support can be used to modify multiple lifestyle behaviors. PMID- 15823787 TI - Efficacy of sequential or simultaneous interactive computer-tailored interventions for increasing physical activity and decreasing fat intake. AB - BACKGROUND: Little evidence exists about the effectiveness of "interactive" computer-tailored interventions and about the combined effectiveness of tailored interventions on physical activity and diet. Furthermore, it is unknown whether they should be executed sequentially or simultaneously. PURPOSE: The purpose of this study was to examine (a) the effectiveness of interactive computer-tailored interventions for increasing physical activity and decreasing fat intake and (b) which intervening mode, sequential or simultaneous, is most effective in behavior change. METHODS: Participants (N = 771) were randomly assigned to receive (a) the physical activity and fat intake interventions simultaneously at baseline, (b) the physical activity intervention at baseline and the fat intake intervention 3 months later, (c) the fat intake intervention at baseline and the physical activity intervention 3 months later, or (d) a place in the control group. RESULTS: Six months postbaseline, the results showed that the tailored interventions produced significantly higher physical activity scores, F(2, 573) = 11.4, p < .001, and lower fat intake scores, F(2, 565) = 31.4, p < .001, in the experimental groups when compared to the control group. CONCLUSIONS: For both behaviors, the sequential and simultaneous intervening modes showed to be effective; however, for the fat intake intervention and for the participants who did not meet the recommendation in the physical activity intervention, the simultaneous mode appeared to work better than the sequential mode. PMID- 15823788 TI - A longitudinal study of exercise barriers in colorectal cancer survivors participating in a randomized controlled trial. AB - BACKGROUND: The Colorectal Cancer and Home-Based Physical Exercise (CAN-HOPE) trial compared the effects of a 16-week home-based exercise program to usual care on quality of life in colorectal cancer survivors. PURPOSE: In this study, we report the weekly exercise barriers from the exercise group. METHODS: Participants randomized to the exercise group (N = 69) were asked to report their exercise on a weekly basis by telephone. Those participants not achieving the minimum weekly exercise prescription (3 times per week of 30 min of moderate intensity exercise) were asked for a primary exercise barrier. RESULTS: We obtained 1,073 (97.2%) weeks of adherence data out of a possible 1,104 (i.e., 69 x 16). Participants did not meet the minimum exercise prescription in 39.2% (421/1,073) of the weeks. We obtained an exercise barrier in 83.8% (353/421) of these cases. Overall, participants reported 37 different exercise barriers; the three most common were lack of time/too busy, nonspecific treatment side effects, and fatigue. The top 7 to 10 barriers accounted for 70% to 80% of all missed exercise weeks. CONCLUSIONS: These findings may have utility for promoting exercise in this population both inside and outside of clinical trials. PMID- 15823789 TI - Calcified tissue histochemistry: from microstructures to nanoparticles. AB - It has long been recognized that histochemistry and cytochemistry offer the only ways of gathering information about the biochemical composition of tissues and cells without disrupting their microscopic architecture. A variety of methods have been put forward for studying nuclei acids, proteins, carbohydrates, lipids, enzymes and other components of intact tissues and cells. By now, many of these have only a historical interest. Some do, however, survive in microscopic and ultramicroscopic applications, and have become incorporated in the most refined and precise techniques that are currently available. Histochemical reactions range from the classic procedures carried out on histological sections to yield final stained products recognizable under the light microscope, to those which are applied on ultrathin sections, using heavy metals or other electron-dense compounds to reveal specific components under the electron microscope; others range from procedures based on the antigen-antibody reaction that are capable of revealing the presence of specific biological molecules, to the biophysical techniques which permit the qualitative and quantitative analysis of elements; lastly, there are the recently proposed ultra-high resolution methods that allow nanoparticles to be recognized. This brief review, which is based on personal experience and on the data in the literature, will discuss the most important methods now being used. PMID- 15823790 TI - The DNA-instability test as a specific marker of malignancy and its application to detect cancer clones in borderline malignancy. AB - Recent progress in cytogenetic and biochemical mutation assay technologies has enabled us to detect single gene alterations and gross chromosomal rearrangements, and it became clear that all cancer cells are genetically unstable. In order to detect the genome-wide instability of cancer cells, a new simple method, the DNA-instability test, was developed. The methods to detect genomic instability so far reported have only demonstrated the presence of qualitative and quantitative alterations in certain specific genomic loci. In contrast to these commonly used methods to reveal the genomic instability at certain specific DNA regions, the newly introduced DNA-instability test revealed the presence of physical DNA-instability in the entire DNA molecule of a cancer cell nucleus as revealed by increased liability to denature upon HCl hydrolysis or formamide exposure. When this test was applied to borderline malignancies, cancer clones were detected in all cases at an early-stage of cancer progression. We proposed a new concept of "procancer" clones to define those cancer clones with "functional atypia" showing positivities for various cancer markers, as well as DNA-instability testing, but showing no remarkable ordinary "morphological atypia" which is commonly used as the basis of histopathological diagnosis of malignancy. PMID- 15823791 TI - The assessment of proliferating cell nuclear antigen immunostaining in myelodysplastic syndromes and its prognostic significance. AB - Several prognostic factors for patients with myelodysplastic syndromes (MDS) have been identified in previous years. In order to determine prognostic factors characterizing haematopoietic cell kinetics, bone marrow proliferative activity and serum TNF-a levels were measured in 51 cases of MDS. Cell proliferation was evaluated by employing a monoclonal antibody directed against the proliferating cell nuclear antigen (PCNA). The PCNA proliferating index (PCNA PI) and serum TNF a levels showed significant differences between patients with MDS and normal controls (p<0.0001). PCNA PI and serum TNF-a were significantly higher in the high risk for leukemic transformation FAB subgroups (RAEB, RAEB-t and CMML) in comparison to the low risk group (RA and RARS) (p<0.001). PCNA PI and TNF-a also increased with increasing IPSS score (p<0.05). A positive correlation was noted between TNF-a concentrations and PCNA PI (r:0.36, p<0.008). Univariate analysis using the log-rank test showed that a higher PCNA PI was associated with a significantly shorter survival (p<0.001). We conclude that elevated PCNA PI and TNF-a serum levels are increased in high risk myelodysplastic disease and that a high PCNA PI is predictive of a shorter survival in this group of patients. PMID- 15823792 TI - Immunolocalization of lactoferrin in surgically resected pigmented skin lesions. AB - Lactoferrin (Lf) expression was determined immunohistochemically in 57 formalin fixed paraffin-embedded bioptic samples obtained from an equal number of patients treated by surgery to remove pigmented skin lesions (nevi = 23; melanoma = 12; vulgaris and seborrhoeic warts = 12; basal cell carcinoma = 10); in addition, 10 specimens of normal skin were studied as control. On 3 mm thick sections, depigmentation and antigen retrieval procedures were performed. The Lf immunoreactivity was revealed by a rabbit anti-human Lf. Quantification of Lf immunoreactivity was performed using an intensity-distribution (ID) score. Melanocytic cells, regardless of their benign or malignant nature, were consistently stained, with no significant differences in the Lf ID-score between melanomas or nevi. A different intensity of Lf immunoreactivity was encountered in superficial portions of warts, exclusively inside squamous epithelial cells arranged in sheets or whorls of keratin. On the contrary, basal cell carcinomas were always unstained, while a slight Lf positivity was found in focal keratinized areas present in two tumours showing baso-squamous differentiation. The Lf immunoreactivity was localized in the cytoplasm and only occasionally in the nucleus. The biological meaning of Lf in these cases of human skin specimens remains unexplained, although it cannot be ruled out that Lf might be involved in the defense system against tumours, or alternatively, may be used by cells requiring iron availability for their turnover. Moreover, the immunohistochemical expression of Lf in melanocytic lesions might be also related to a Lf-melanin interaction. Finally, the involvement of Lf in skin squamous non-neoplastic elements could be related to its role as one of the molecules modulating an unspecific inflammatory or anti-oxidant response. PMID- 15823793 TI - Phenotype commitment in vascular smooth muscle cells derived from coronary atherosclerotic plaques: differential gene expression of endothelial nitric oxide synthase. AB - Unstable angina and myocardial infarction are the clinical manifestations of the abrupt thrombotic occlusion of an epicardial coronary artery as a result of spontaneous atherosclerotic plaque rupture or fissuring, and the exposure of highly thrombogenic material to blood. It has been demonstrated that the proliferation of vascular smooth muscle cells (VSMCs) and impaired bioavailabilty of nitric oxide (NO) are among the most important mechanisms involved in the progression of atherosclerosis. It has also been suggested that a NO imbalance in coronary arteries may be involved in myocardial ischemia as a result of vasomotor dysfunction triggering plaque rupture and the thrombotic response. We used 5' nuclease assays (TaqMan PCRs) to study gene expression in coronary plaques collected by means of therapeutic directional coronary atherectomy from 15 patients with stable angina (SA) and 15 with acute coronary syndromes (ACS) without ST elevation. Total RNA was extracted from the 30 plaques and the cDNA was amplified in order to determine endothelial nitric oxide synthase (eNOS) gene expression. Analysis of the results showed that the expression of eNOS was significantly higher (p<0.001) in the plaques from the ACS patients. Furthermore, isolated VSMCs from ACS and SA plaques confirmed the above pattern even after 25 plating passages. In situ RT-PCR was also carried out to co-localize the eNOS messengers and the VSMC phenotype. The eNOS gene was more expressed in ACS plaques and VSMCs cultured from them, thus indicating that: a) the expression of the most important differentiation markers is retained under in vitro conditions; and b) NO may play a pivotal role in coronary artery disease. Our findings suggest a new cell system model for studying the pathophysiology of unstable angina and myocardial infarction. PMID- 15823794 TI - Multicolor fluorescence technique to detect apoptotic cells in advanced coronary atherosclerotic plaques. AB - Apoptosis occurring in atherosclerotic lesions has been suggested to be involved in the evolution and the structural stability of the plaques. It is still a matter of debate whether apoptosis mainly involves vascular smooth muscle cells (vSMCs) in the fibrous tissue or inflammatory (namely foam) cells, thus preferentially affecting the cell-poor lipid core of the atherosclerotic plaques. The aim of the present investigation was to detect the presence of apoptotic cells and to estimate their percentage in a series of atherosclerotic plaques obtained either by autopsy or during surgical atherectomy. Apoptotic cells were identified on paraffin-embedded sections on the basis of cell nuclear morphology after DNA staining and/or by cytochemical reactions (TUNEL assay, immunodetection of the proteolytic poly (ADP-ribose) polymerase-1 [PARP-1] fragment); biochemical procedures (identifying DNA fragmentation or PARP-1 proteolysis) were also used. Indirect immunofluorescence techniques were performed to label specific antigens for either vSMCs or macrophages (i.e., the cells which are most likely prone to apoptosis in atherosclerotic lesions): the proper selection of fluorochrome labeling allowed the simultaneous detection of the cell phenotype and the apoptotic characteristics, by multicolor fluorescence techniques. Apoptotic cells proved to be less than 5% of the whole cell population, in atherosclerotic plaque sections: this is, in fact, a too low cell fraction to be detected by widely used biochemical methods, such as agarose gel electrophoresis of low-molecular-weight DNA or Western-blot analysis of PARP-1 degradation. Most apoptotic cells were of macrophage origin, and clustered in the tunica media, near or within the lipid rich core; only a few TUNEL-positive cells were labeled for antigens specific for vSMCs. These results confirm that, among the cell populations in atherosclerotic plaques, macrophage foam-cells are preferentially involved in apoptosis. Their death may decrease the cell number in the lipid core and generate a possibly defective apoptotic clearance: the resulting release of matrix-degrading enzymes could contribute to weakening the fibrous cap and promote the plaque rupture with the risk of acute ischemic events, while increasing the thrombogenic pultaceous pool of the plaque core. PMID- 15823795 TI - High-level detection of gene amplification and chromosome aneuploidy in extracted nuclei from paraffin-embedded tissue of human cancer using FISH: a new approach for retrospective studies. AB - A novel application of fluorescence in situ hybridization (FISH) to isolated nuclei is described. The method detects gene amplification and chromosome aneuploidy in extracted nuclei from paraffin-embedded tissue of human cancer with greater sensitivity and specificity than existing FISH methods. In this study, the method is applied to signal detection of the HER-2/neu (c-erbB-2) gene, whose amplification is one of the most common genetic alterations associated with human breast cancer. Nuclei were extracted and isolated from formalin fixed, paraffin embedded tissue of 43 different carcinomas (breast, ovary, endometrium, gastrointestinal stromal tumor and malignant mesothelioma). FISH was performed both on sections and extracted nuclei of each tissue using chromosome enumeration probes (CEP) for the centromeric regions of chromosomes 8 and 17, and a locus specific identifier (LSI) for the HER-2/neu oncogene. Differences between ploidy calculated in sections and extracted nuclei were seen in 3 breast carcinomas and 1 gastrointestinal stromal tumor (GIST). Furthermore, 1 breast cancer, previously considered to be borderline for HER-2/neu gene amplification turned out to be clearly amplified. Nuclei extraction and isolation bypass all the problems related to signal interpretation in tissue sections, and the adoption of this new technique, which improves the signal quality in several neoplastic samples, is suggested. PMID- 15823796 TI - The expression of aromatase, estrogen receptor alpha and estrogen receptor beta in mouse Leydig cells in vitro that derived from cryptorchid males. AB - A broad expression of aromatase and estrogen receptors (ERs) in the testis suggests an important role for estrogens in regulating testicular cell function and reproductive events. The aim of the present study was to show whether Leydig cells in vitro isolated from cryptorchid testes of two inbred strains of mice, KE and CBA, are a site of estrogen synthesis. Using immunocytochemistry, aromatase, estrogen receptor alpha(ERalpha), and estrogen receptor beta(ERbeta) were localized in cultured Leydig cells. Immunoreactive aromatase was found in the cytoplasm of control Leydig cells and those isolated from cryptorchid males, however the intensity of immunostaining was different, being stronger in Leydig cells deriving from cryptorchid mice. The strongest aromatase immunostaining was found in cryptorchid-KE Leydig cells. Strong immunoexpression of ERalpha was detected in the nuclei of both KE-and CBA-Leydig cells. The intensity of ERalpha immunostaining was stronger in cultured cells deriving from cryptorchid testes. ERbeta immunoexpression was detected predominantly in KE-Leydig cells. Control CBA-Leydig cells were negative for ERbeta or the result was inconclusive, whereas in cryptorchid CBA-Leydig cells a weak immunostaining was present in their nuclei. Western blot analysis confirmed the results obtained by immunocytochemistry. In KE- and CBA-Leydig cells aromatase as a band of 55 kDa protein was present, whereas ERalpha molecular weight was 67 kDa on Western blots. No band was detected for ERbeta. Radioimmunological analysis revealed that androgen and estrogen levels secreted by Leydig cells in vitro were strain dependent. Additionally, in KE-Leydig cells that derived from cryptorchid mice estrogen level was distinctly higher in comparison with that of the respective control. PMID- 15823797 TI - Differentiation of monocytic U937 cells under static magnetic field exposure. AB - We present here a morphological, cytochemical and biochemical study of the macrophagic differentiation of human pro-monocytic U937 cells exposed to moderate intensity (6 mT) static magnetic fields (MF). It was found that the following substances induced differentiation in U937 cells to a progressively lower degree: 50 ng/mL 12-0-tetradecanoyl-13-phorbol acetate (TPA), low concentration of glutamine (0,05 mM/L), 10% dimethyl sulfoxide (DMSO) and 100 mM/L Zn++. Differentiated U937 cells shift from a round shape to a macrophage-like morphology, from suspension to adhesion growth and acquire phagocytotic activity, the cytoskeleton adapting accordingly. Exposure to static MF at 6 mT of intensity decreases the degree of differentiation for all differentiating molecules with a consequent fall in cell adhesion and increased polarization of pseudopodia and cytoplasmic protrusions. Differentiation alone, or in combination with exposure to static MFs, affects the distribution and quantity of cell surface sugar residues, the surface expression of markers of macrophage differentiation, and phagocytotic capability. Our results indicate that moderate-intensity static MFs exert a considerable effect on the process of macrophage differentiation of pro monocytic U937 cells and suggest the need for further studies to investigate the in vivo possible harmful consequences of this. PMID- 15823798 TI - Immunocytochemical study of estrogen receptor activation factor (E-RAF) and the proteins that interact with nuclear estrogen receptor II (nER II) in epithelial endometrial cells, in the presence and in the absence of estradiol. AB - The localization and abundance of the estrogen receptor activation factor (E-RAF) and a small nuclear ribonucleoprotein (snRNP) complex containing three proteins, p32, p55 and p60, which interact with the nuclear estrogen receptor II (nER II), have been studied in rat endometrial epithelial cells by means of immunofluorescence and high resolution quantitative immunocytochemistry. In the cytoplasm E-RAF is associated with the rough endoplasmic reticulum. In the nucleus it is mainly localized at the interchromatin space, and surrounding the clumps of compact or semi-condensed chromatin. Quantitative analyses show that the abundance of E-RAF in the nucleus increases after ovariectomy and decreases 3 minutes after estradiol administration. These results are in agreement with the currently available biochemical data. Double immunolocalizations demonstrate that p32, p55, p60 co-localize with other splicing-related protein. High resolution immunolocalization shows that p32, p55, p60 are associated with perichromatin fibrils (co-transcriptional splicing) and with clusters of interchromatin granules (storage of splicing-related molecules). The nuclear abundance of the snRNP complex decreases with ovariectomy, increases within 3 minutes after estradiol administration and remains higher than that in ovariectomized animals for 27 minutes. These results strongly support the previous data on the role of nER-II in the regulation of mRNA transcription and its export from the nucleus to the cytoplasm. PMID- 15823799 TI - Tachykinin neuropeptides in cerebellar granule neurons: an immunocytochemical study. AB - We previously demonstrated that exogenously administered neurokinin A and neurokinin B, but not substance P, increased the sensitivity of cultured cerebellar granule neurons (CGNs) to glutamate. In the present study, the presence of tachykinin neuropeptides in CGNs was tested by confocal-based immunofluorescence. We found that neurokinin A and neurokinin B are present in CGNs but absent in astrocytes while substance P is abundant in astrocytes but absent in CGNs. It is postulated that the different localization of tachykinin neuropeptides in CGNs and astroglial cells has a physiological role in the modulation of excitatory transmission. PMID- 15823800 TI - Advanced microscopic and histochemical techniques: diagnostic tools in the molecular era of myology. AB - Over the past two centuries, myology (i.e. the basic and clinical science of muscle and muscle disease) has passed through 3 stages of development: the classical period, the modern stage and the molecular era. The classical period spans the last part of nineteenth century and the earlier part of the twentieth century. During this time, several major muscle diseases were clinically and pathologically characterized, including Duchenne muscular dystrophy (DMD), myotonic dystrophy (DM) and facioscapulohumeral dystrophy (FSHD). The modern stage in the second half of the twentieth century is characterized by the adaptation of histo and cytochemical techniques to the study of muscle biopsies. These tools improved the diagnostic accuracy and made possible the identification of new changes and structures (Engel and Cunningham, 1963; Scarlato, 1975). PMID- 15823801 TI - Histochemistry and translational medicine. PMID- 15823802 TI - Cochlear hair cell loss in single-dose versus continuous round window administration of gentamicin. AB - CONCLUSIONS: Gentamicin-induced cochlear hair cell loss depends on local middle ear administration kinetics and the total drug dose. Single-dose gentamicin instillation in the middle ear is associated with a high variation in hair cell loss. OBJECTIVE: To compare the effects of single-dose and continuous round window administration of gentamicin on cochlear hair cell loss in a guinea pig model. MATERIAL AND METHODS: Two methods for drug administration to the inner ear were used. In groups of five animals, a total dose of 0.8 or 3.2 mg of gentamicin was either instilled as a single dose directly into the round window niche or administered continuously over a 1-week period using a pump-catheter system. Continuous administration was achieved by means of a posterior tympanotomy and subcutaneous placement of an osmotic pump fitted with a catheter. The tip of the catheter was fixed in the round window niche. One group of five animals served as controls and received a saline infusion. The animals were sacrificed after 1 week and hair cell loss was determined microscopically after dissection and phalloidin labelling of the basilar membrane and organ of Corti. RESULTS: Quantitation of cochlear hair cell loss revealed a dose-dependent effect of gentamicin. With both treatment modalities the higher dose induced a higher percentage of hair cell loss. There was inner and outer hair cell loss in all four groups that received gentamicin. With the single-dose instillation, hair cell loss was distributed irregularly from the round window membrane towards the cochlear apex, whereas continuous administration induced hair cell loss close to the round window membrane. Single-dose instillation induced greater hair cell loss than continuous administration at the same dose. The inter-individual variation in hair cell loss was highest following single-dose instillation. PMID- 15823803 TI - External otitis caused by infection with Pseudomonas aeruginosa or Candida albicans cured by use of a topical group III steroid, without any antibiotics. AB - CONCLUSIONS: Irrespective of the microbial agent, group III steroid solution cured external otitis efficiently in a rat model. The addition of antibiotic components to steroid solutions for the treatment of external otitis is of questionable validity. OBJECTIVE: External otitis, caused by infection with either Pseudomonas aeruginosa or Candida albicans, was established in a rat model and the treatment efficacy of a group III steroid solution was studied. MATERIAL AND METHODS: Three treatments were studied: (i) a group III steroid solution; (ii) a group I steroid combined with two antibiotic components; and (iii) a saline solution. A scoring scale was used to evaluate the characteristics of the ear canal skin. Bacteriological and fungal samples were collected for culturing and ear canal skin biopsies were taken for structural analyses. RESULTS: It was possible to cause P. aeruginosa and C. albicans infections in an animal model. In the P. aeruginosa-infected animals, only the group III steroid treatment cured all the animals. In the C. albicans-infected animals, group III steroid treatment resolved external otitis faster than the other treatment modalities. PMID- 15823804 TI - Dynamic visual acuity using "far" and "near" targets. AB - CONCLUSIONS: DVA may be useful for assessing the functional consequences of an impaired gaze stabilization mechanism or for testing the effectiveness of a rehabilitation paradigm. Because target distance influences the relative contributions of canal and otolith inputs, the ability to measure DVA at near and far viewing distances may also lead to tests that will independently assess canal and otolith function. OBJECTIVE: To present and test a methodology that uses dynamic visual acuity (DVA) to assess the efficacy of compensatory gaze mechanisms during a functionally relevant activity that differentially measures canal and otolith function. MATERIAL AND METHODS: The effect of treadmill walking at a velocity of 1.79 m/s on subjects' visual acuity was assessed at each of two viewing distances. A custom-written threshold determination program was used to display Landolt C optotypes on a laptop computer screen during a "far" (4 m) target condition and on a micro-display for a "near" (50 cm) target condition. The walking acuity scores for each target distance were normalized by subtracting a corresponding acuity measure obtained while standing still on the treadmill belt. RESULTS: As predicted by subjective reports of relative target motion, the decrease in visual acuity was significantly greater (p < 0.00001) for the near compared to the far condition. PMID- 15823805 TI - Blink reflex and auditory speech perception in prelingually cochlear-implanted children. AB - CONCLUSION: The results of this study show that R- prelingually cochlear implanted children are not optimal candidates for cochlear implantation. However, if this group are implanted, other rehabilitation methods should be incorporated into their rehabilitation program in order to achieve better results and maximize the efficacy of their prosthesis. OBJECTIVE: To demonstrate that the blink reflex or auropalpebral reflex evaluation can be used as a prognostic factor for the assessment of auditory and speech perception levels in prelingually cochlear implanted children. MATERIAL AND METHODS: In an observational, analytical, prospective study conducted at a single cochlear implant rehabilitation center in 85 prelingually cochlear-implanted children, the presence or absence of the blink reflex (BR) was evaluated and the results of auditory and speech perception tests were compared between reflex-positive (R+) and reflex-negative (R-) patients. To obtain the BR, four electrodes were applied in both the Nucleus and MED-EL systems and then stimulated by means of current levels higher than the previously detected most comfortable level of the patient until the reflex appeared. Auditory and speech perception levels were measured using the vowels-confusion test and the categorization of auditory perception scale. RESULTS: The mean results of auditory and speech perception tests were significantly higher in R+ compared to R- patients. PMID- 15823806 TI - Pre- and intraoperative predictive factors of facial palsy in vestibular schwannoma surgery. AB - CONCLUSION: S These results support previous ones with regard to FN risk factors in VS surgery. However, they also provide new preoperative factors that influence postoperative FN function, such as clinical symptoms, the nature of the surgical procedure (use of laser) and ABR results. OBJECTIVE: To determine pre- and perioperative factors influencing facial nerve (FN) outcome in vestibular schwannoma (VS) surgery. MATERIAL AND METHODS: A total of 424 patients undergoing VS surgery were included in this retrospective study. Patients were divided into two groups according to the existence or absence of a FN palsy during the 8 days following surgery (Groups 1 and 2, respectively). Various parameters were evaluated preoperatively as follows. Quantitative parameters: age; duration of clinical symptoms; pure-tone audiometry (PTA) results; speech reception threshold; speech discrimination score; auditory brainstem response (ABR) results; and transient-evoked otoacoustic emission amplitude. Qualitative parameters: gender; side of the tumor; angle between the tumor and the internal auditory canal (VS/IAC angle) < or = or > 30 degrees; MRI aspect (n = 69); surgical approach; ease of the surgical procedure, the use or non-use of laser dissection; and the histological Antoni's type of the tumor. RESULTS Pre- and perioperative factors that differed significantly between Groups 1 and 2 were as follows. Quantitative factors: tinnitus duration was longer and PTA and ABR results were worse in Group 1. Qualitative factors: heterogenous/cystic MRI aspect, use of retrosisgmoid and middle fossa approaches, easy surgical procedure, dissection without laser and Antoni's type A were more frequently found in Group 1. PMID- 15823807 TI - Quantitative expression levels of regulated on activation, normal T cell expressed and secreted and eotaxin transcripts in toluene diisocyanate-induced allergic rats. AB - CONCLUSION: These results suggest that eotaxin may play a predominant role in controlling antigen-induced eosinophil recruitment into tissue. Objective To investigate the expression levels of regulated on activation, normal T cell expressed and secreted (RANTES) mRNA and eotaxin mRNA in the nasal mucosa of toluene diisocyanate (TDI)-induced allergic rats and to evaluate which of them is primarily related to selective eosinophilic infiltration by comparing their expression levels with the numbers of infiltrated eosinophils and vascular cell adhesion molecule-1 (VCAM-1). MATERIAL AND METHODS: We quantified the expression levels of two strong eosinophilic CC chemokines (RANTES and eotaxin) and VCAM-1 at mRNA levels in the nasal mucosa of TDI-induced allergic rats using competitive polymerase chain reaction and compared their expression levels with the number of infiltrated eosinophils. RESULTS: The number of infiltrated eosinophils was significantly increased between 3 h and Day 4 in TDI-induced allergic rats, but had decreased by Day 5. VCAM-1 mRNA expression was also increased between 3 h and Day 4. The number of infiltrated eosinophils correlated with the expression levels of VCAM-1 mRNA (p < 0.01). In contrast, expression of RANTES mRNA and eotaxin mRNA was increased between 3 h and Day 2, peaked between Days 1 and 2 and then declined. Although the expression of both chemokines correlated with the numbers of infiltrated eosinophils (p < 0.01), peak expression levels of eotaxin mRNA were 14-fold higher than baseline levels whereas RANTES mRNA expression increased 3-fold. PMID- 15823808 TI - Significance of endogenous glucocorticoid sensitivity for airway eosinophilia in a murine model of allergy. AB - CONCLUSIONS: Endogenous GC protects against allergic inflammatory responses in the airways. These effects are modulated by both peripheral blockade and inhibition of release. Individual response patterns to stress, i.e. corticosterone release and peripheral sensitivity, may influence both the central and peripheral levels of the allergic airway reaction in patients. OBJECTIVE: Glucocorticoids (GCs) modulate the allergic inflammatory response. Acute or chronic stress will influence circulating levels of GCs, rates of secretion, metabolism and target tissue sensitivity. In a clinical situation, stress may exacerbate or attenuate the asthmatic reaction. The aim of this study was to investigate the effects of inhibition of endogenous GC in an allergic airway inflammation model in the mouse. MATERIAL AND METHODS: An ovalbumin model using i.p. sensitization and intra-nasal challenge was used for respiratory eosinophilic inflammation. GC release was inhibited by administration of metyrapone (ME), and peripheral glucocorticoid receptors were blocked by administration of RU486 (RU). RESULTS: Inhibition with RU and ME increased eosinophilia in the bone marrow compared to controls (p < 0.05). Eosinophilia in bronchoalveolar lavage fluid increased in the sensitized groups compared to controls, but there were no differences between the sensitized groups. CD3+ and CD4+ cells were increased in the nasal mucosa as a result of treatment with RU and ME. PMID- 15823809 TI - Valproic acid-associated sialadenosis of the parotid and submandibular glands: diagnostic and therapeutic aspects. AB - Sialadenosis has been defined as a non-inflammatory, parenchymatous salivary gland disease causing recurrent, bilateral swelling of the salivary glands. As an adverse drug reaction of valproic acid, sialadenosis is very rare. To our knowledge, it has been reported only once in the world literature to date. We present herein the case of a patient with valproic acid-associated sialadenosis of both the parotid and submandibular glands. This appears to be the first published case of a patient who received surgical treatment. On light and electron microscopy of all the affected salivary glands, granular sialadenosis with predominantly moderate electron-dense secretory cytoplasmatic granules was observed. No relevant degenerative alterations were seen. There was no histological evidence of peripheral neuropathy of the nerve supply, leading to disordered activity of acinar cells by loss of neurosecretory granules. Lateral parotidectomy, performed under neuromonitoring control for safety reasons, is the treatment of choice for chronic recurrent parotitis that does not respond to conservative therapy, particularly if the cosmetic deformity is unacceptable to the patient. If the submandibular glands are involved, partial removal is recommended. PMID- 15823810 TI - Soluble CD44 standard, CD44 variant 5 and CD44 variant 6 and their relation to staging in head and neck cancer. AB - CONCLUSION: The possible roles of CD44st, CD44v5 and CD44v6 in the prognosis of head and neck cancer deserve further elucidation and evaluation with long-term patient follow-up. OBJECTIVE: Standard CD44 (CD44st), CD44 variant 5 (CD44v5) and CD44 variant 6 (CD44v6) are expressed in human malignant cells and tissues. The mechanism of their expression remains unclear, but has been reported to be associated with the progression and metastasis of malignancies. Recently, it has frequently been reported that the prognosis of head and neck cancer is associated with expression of the cell adhesion molecule CD44. MATERIAL AND METHODS: We investigated correlations between the soluble adhesion molecule CD44 and clinicopathologic variables, for example, age, sex, histologic grade, tumor size, lymph node status, distant metastasis and TNM stage. The pre- and post-treatment serum levels of CD44st, CD44v5 and CD44v6 were determined by means of ELISAs in 81 patients with head and neck cancer and 20 healthy volunteers (controls). RESULTS: In the cancer patients, the pre-treatment median serum levels of CD44st, CD44v5 and CD44v6 were 327 +/- 134, 312 +/- 118 and 211 +/ 110 ng/ml, respectively. The corresponding post-treatment levels were 185 +/- 103, 177 +/- 90 and 110 +/- 65 ng/ml. In the healthy volunteers, the median serum levels of CD44st, CD44v5 and CD44v6 were 133 +/- 40, 142 +/- 39 and 86 +/- 22 ng/ml, respectively. In the cancer patients, there was no significant correlation between the serum levels of CD44st, CD44v5 and CD44v6 and the clinicopathological variables. The pre-treatment serum levels of CD44st, CD44v5 and CD44v6 were closely associated with TNM stage (p = 0.0017, 0.0005 and 0.0046, respectively). The median pre-treatment serum levels of CD44st, CD44v5 and CD44v6 were significantly higher than those in the control group (p = 0.0002, 0.0065 and 0.0038, respectively). The median post- treatment serum levels of CD44st, CD44v5 and CD44v6 were significantly lower than the pre-treatment levels (p = 0.0003, 0.0027 and 0.0034, respectively). PMID- 15823811 TI - Laryngeal and cranial nerve involvement after carotid endarterectomy. AB - CONCLUSIONS: Laryngeal and/or cranial nerve involvement after CEA surgery is not a rare condition, occurring in almost half of operated subjects. However, in most cases the functional deficit is transient and does not need any particular form of treatment. In this study, specific rehabilitative procedures were needed in only a relatively small number of cases (9%). A routine ENT examination has also proved to be extremely useful for detecting slight functional deficits which may occur following CEA surgery, bearing in mind that possible permanent lesions may require a rehabilitative procedure. OBJECTIVE: To identify, by means of a careful otolaryngologic examination, the incidence and degree of cranial nerve deficit related to carotid endarterectomy (CEA), starting from the first postoperative days. MATERIAL AND METHODS: A consecutive cohort of patients with symptomatic and asymptomatic carotid artery stenosis who underwent CEA was carefully followed on the basis of possible laryngeal and/or cranial nerve involvement. An ENT examination was carried out preoperatively (phase I) and at different times [3 (phase II) and 15 days (phase IIIa)] after surgery; in addition, patients with persisting neurological lesions were also checked 60 days after surgery (phase IIIb). RESULTS: In 59% of the patients, isolated or associated forms of deficit were found. Only 17.5% of these deficits did not appear to be transient, but rehabilitative procedures for voice or swallowing impairments were only needed in 9% of them. PMID- 15823812 TI - Intraoperative lymphatic mapping in cases of midline squamous cell carcinoma. AB - OBJECTIVE: To analyze the value of intraoperative lymphatic mapping in cases of midline primary head and neck squamous cell carcinoma (HNSCC) in clinically staged N0 necks. MATERIAL AND METHODS: Eleven patients with HNSCC of the epiglottis (2 T1, 6 T2, 3 T3), all of whom were staged with a neck status of N0 using sonography and CT, underwent intraoperative peritumoral (99m)Tc-nanocoll injection (4 sites; 45 MBq), radiolabeled detection and analysis of up to 3 hot sentinel nodes (SNs) during elective neck dissection. RESULTS: Gamma probe use revealed bi- and unilateral intranodal tracer uptake in 6/11 and 5/11 patients, respectively. In 2/6 patients with bilateral intranodal tracer uptake an SN with an isolated metastasis was found at one neck site while the other four patients were tumor-free in the SNs. Of the five patients with unilateral intranodal tracer uptake, three had radiolabeled SNs containing isolated metastases whereas two had no cancer detected, giving a total occult cancer rate of 45% (5/11). No cancer was found in non-labeled nodes. CONCLUSIONS: Intraoperative lymphatic mapping correctly identified the stage of metastatic disease. Unilateral tracer uptake represented the pathway of occult metastatic spread in 3/5 patients and the disease-free neck status of both neck sites in 2/5 patients. No patient had occult bilateral cancer. Future investigations should be done to determine whether intraoperative lymphoscintigraphy can guide the indication for unilateral only or bilateral neck dissection in these patients. PMID- 15823813 TI - Human telomerase reverse transcriptase expression and its clinical significance in laryngeal squamous cell carcinoma. AB - CONCLUSIONS: Upregulation of hTERT mRNA plays an important role during the occurrence and development of laryngeal squamous cell carcinoma. Most hTERT mRNA in plasma from patients with laryngeal squamous cell carcinoma is derived from tumour cells; moreover, the determination of plasma hTERT mRNA contributes to tumour diagnosis and the observation of curative effect. OBJECTIVES: To establish a real-time fluorescent reverse transcriptase polymerase chain reaction (RT-PCR) and to quantitate the level of human telomerase reverse transcriptase (hTERT) mRNA in carcinoma tissue and plasma from patients with laryngeal squamous cell carcinoma. We also wished to evaluate the role that hTERT mRNA expression plays during the occurrence and development of laryngeal squamous cell carcinoma, to probe the correlation between the expression level and the clinical and pathological parameters and to investigate the value of the determination of plasma hTERT mRNA in tumour diagnosis and the observation of curative effect. MATERIAL AND METHODS: A real-time fluorescent RT-PCR and a Lightcycler PCR system were used to quantitate the expression level of hTERT mRNA. RESULTS: The expression levels of hTERT mRNA (NhTERT) from laryngeal squamous cell carcinoma tissue and corresponding adjacent non-cancerous tissue were 62.6 +/- 21.7 and 3.5 +/- 1.9, respectively. NhTERT was significantly elevated in laryngeal squamous cell carcinoma tissue, rising to 17.9-fold on average, but there was no significant correlation between NhTERT and either tumour location, differentiation degree, T grade or N grade. For healthy examinees, NhTERT in plasma was 1.3 +/- 0.9, compared to 13.1 +/- 9.4 and 9.3 +/- 5.8 in patients with laryngeal squamous cell carcinoma examined before and 2 days after surgery, respectively. Compared to healthy examinees, NhTERT in plasma from patients with laryngeal squamous cell carcinoma was significantly elevated; moreover, 2 days after surgery, NhTERT in plasma had decreased significantly in these patients. PMID- 15823814 TI - Human papillomaviruses in lymph node neck metastases of head and neck cancers. AB - CONCLUSION: The results of this study corroborate earlier findings that human papillomavirus (HPV)16 is the most prevalent type of HPV in squamous cell carcinomas of the head and neck (SCCHNs) and reinforce a possible influence of HPV on SCCHN progression by showing that the majority of HPV-positive patients harbor HPV16 (or HPV33) both in their primary tumors and in lymph node neck metastases (LNNMs). OBJECTIVE: HPVs are causally associated with carcinomas of the uterine cervix and have also been linked to a subset of SCCHNs. In order to further investigate the predicted causative role of HPV in SCCHNs, we analyzed pairs of primary tumors and LNNMs or LNNMs alone for the presence of HPV DNA using polymerase chain reaction (PCR). MATERIAL AND METHODS: DNA was extracted from fresh frozen tissue samples of primary tumors and the corresponding LNNMs of 18 patients and from LNNMs alone in 17 patients. For the detection and typing of HPV, PCR was performed using both type-specific and consensus primer pairs, followed by Southern hybridization and, in selected cases, sequencing of the PCR products. RESULTS: Of the 35 patients investigated, 22 (63%) were found to have HPV DNA in their tumors: HPV16 DNA in 21 cases and HPV33 in 1. The highest HPV prevalence was detected in tumors of Waldeyer's tonsillar ring (8/9 patients; 89%). Of the 18 patients in whom primary tumors and LNNMs were analyzed, 7 (39%) were HPV-positive in both samples (HPV16, n = 6; HPV33, n = 1), in 3 (17%) the primary tumors were HPV-negative and the LNNMs HPV16-positive and in 1 (5.5%) the primary tumor contained HPV16 and the LNNM was negative. Interestingly, of the 7 patients in whom LNNMs had been detected only several months after diagnosis and treatment of the primary tumors, only 1 showed infection with HPV (HPV33). PMID- 15823815 TI - Characterization of mitochondrial respiratory chain energetics in the vestibular nucleus complex. AB - CONCLUSIONS: Despite having very high neuronal firing rates, the VNC does not have unusually high mitochondrial activity in vitro. This study is the first in which functionally active mitochondria from the hindbrain have been isolated and characterized. OBJECTIVE: Neurons in the vestibular nucleus complex (VNC) have exceptionally high spontaneous firing rates. Neuronal mitochondria generate adenosine triphosphate critical for maintaining the membrane potentials required for axon firing. We therefore hypothesized a high rate of mitochondrial activity in the VNC. MATERIAL AND METHODS: To test this hypothesis, we compared mitochondrial activity in the VNC with mitochondrial activity from another area of the hindbrain, the cerebellum. Mitochondrial respiratory activity was assessed by measuring oxidative phosphorylation and mitochondrial respiratory enzyme complex activity. RESULTS: Assay results were not significantly different in the VNC compared to those obtained with the cerebellum or with rat brain mitochondria in previous studies. PMID- 15823816 TI - Peripheral primitive neuroectodermal tumor of the cerebellopontine angle. AB - Peripheral primitive neuroectodermal tumors are highly malignant small cell neoplasms. A 27-year-old female presented with a 6-month history of right-sided facial pain and progressive weakness of the facial muscles. She had non-pulsative tinnitus, progressive right hearing loss and facial palsy. T2-weighted MRI showed a heterogeneous hyperintense lesion invading the right internal auditory canal. Surgical removal was performed. Pathological examination showed sheets of small cells with irregular nuclei. Immunohistochemical studies demonstrated positive immunoreactivity for neuron-specific enolase, synaptophysin, chromogranin, vimentin, S-100 protein and p30-32 MIC-2 gene product. The patient was treated with chemotherapy (etoposide, vincristine, adriamycin, ifosfamide and actinomycin D) and radiotherapy. After 65 months of follow-up, the patient presented with cervical metastasis. Radical cervical dissection was performed and the patient was treated with a second course of chemotherapy. At control MRI after 29 months of follow-up the patient showed no signs of local recurrence or distant metastasis. PMID- 15823817 TI - Lesion site in idiopathic bilateral vestibulopathy: a galvanic vestibular-evoked myogenic potential study. AB - CONCLUSION: The result suggests that patients with idiopathic bilateral vestibulopathy may have nerve lesions when the inferior nerve system is affected, while the inferior vestibular nerve system may be spared. OBJECTIVE: To clarify the lesion site in idiopathic bilateral vestibulopathy, an acquired bilateral vestibulopathy of unknown cause. MATERIAL AND METHODS: Two 75-year-old males diagnosed with idiopathic bilateral vestibulopathy were enrolled. Both showed absent or highly decreased responses on the caloric test on both sides. They underwent vestibular-evoked myogenic potential (VEMP) testing by means of acoustical and electrical stimulation. As acoustic stimulation, 95 dB nHL clicks and short tone bursts (500 Hz) were presented, while 3 mA (1 ms) short-duration galvanic stimuli were presented as electrical stimulation. Responses were recorded on the sternocleidomastoid muscles. RESULTS: Both patients showed unilateral absence of VEMPs with both acoustic and short-duration galvanic stimuli. PMID- 15823818 TI - Inflammatory myofibroblastic tumor of the parotid gland: case report and review of the literature. AB - An inflammatory myofibroblastic tumor, previously known as an inflammatory pseudotumor, is an uncommon neoplasm. This tumor, which has characteristic morphological and immunohistochemical features, is mostly seen in the lung. Herein we present a rare case of an inflammatory myofibroblastic pseudotumor of the parotid gland as well as a review of the literature. The patient was a 66 year-old man with recurrent painful swelling of the parotid gland. A total parotidectomy with preservation of the facial nerve branches was performed. The patient showed no signs of recurrence > 3 years after surgery. The presence of clonal cytogenic abnormalities supported the neoplastic origin of this process. The treatment consisted of complete resection. Clinicians should however be aware that an inflammatory myofibroblastic tumor may mimic a reactive process. PMID- 15823819 TI - Renal metastasis from papillary thyroid microcarcinoma. AB - Papillary or follicular microcarcinoma of the thyroid comprises 10-20% of all thyroid malignancies. Most of these diseases exhibit slow progression and have a favorable prognosis. Distant metastasis caused by thyroid microcarcinoma is uncommon, and is usually found in the lung or bone. Thyroid microcarcinoma with metastasis to the kidney has not previously been reported. Clinically detectable well-differentiated metastatic thyroid carcinoma to the kidney is rare, and only 16 cases have been reported. Herein we describe a case of metastatic papillary thyroid microcarcinoma to the kidney in a patient with a pelvic fracture and pulmonary metastasis. The patient underwent a total thyroidectomy, radiotherapy, ablation treatment with 131I and thyroxine suppressive therapy. In conclusion, microcarcinoma cannot be ignored. Although uncommon, metastases may arise. PMID- 15823820 TI - Traumatic retropharyngeal haematoma in an anticoagulated patient: Case report and proposal for a clinical protocol. AB - Retropharyngeal haematomas (RPHs) are rare but potentially life-threatening conditions that require a prompt diagnosis. However, the clinical scenario is not always straightforward as their presentation may be insidious, with no specific signs or symptoms. Treatment of RPH is conservative in the majority of cases, with close observation. Nevertheless, surgical intervention is sometimes indicated for large, non-resolving haematomas. We present the case of a 53-year old woman on anticoagulant therapy who required evacuation of a traumatic RPH. We also propose a clinical protocol for the management of these entities according to our experience and previous literature reports. PMID- 15823821 TI - Nasal angiomyolipoma. AB - Angiomyolipomas (AMLs) are hamartomatous lesions containing smooth muscle, vasculature and mature adipose tissue. AML occurs most frequently in the kidneys, with the liver being the second commonest site of involvement. Other localizations for AML are extremely rare and, to our knowledge, only six cases of AML of the nasal cavity have been reported to date. Our case involved a 52-year old male who presented with a history of snoring and obstruction of the left nasal cavity. Physical examination revealed a 3-cm polypoid mass in the left nasal cavity, which was excised surgically. Histopathologic examination revealed that the lesion was composed of smooth muscle bundles, mature adipose tissue and blood vessels of different sizes. No epitheloid smooth muscle cells were observed and HMB-45 staining was negative. In conclusion, although nasal AML is a rare lesion, it should be considered in the differential diagnosis of polypoid nasal masses, especially when the mass occurs unilaterally. PMID- 15823822 TI - Lack of association between endometriosis and the CYP17 MspA1 polymorphism in UK and Japanese populations. AB - Endometriosis is a complex trait, which means that multiple susceptibility genes interact with one another and the environment to produce the phenotype. One of the genes previously implicated in the disease is CYP17; this encodes the enzyme P450c17alpha, which plays a vital role in steroid biosynthesis in the ovary. The presence of a single nucleotide polymorphism (T-->C) in the 5'-promoter region of the gene creates a new recognition site for the restriction enzyme MspA1 producing a mutant allele (A2), which affects circulating estrogen levels. In this study, we compared the frequency of the CYP17 MspA1 polymorphism in two different ethnic populations. DNA was obtained from (1) 94 women with revised American Fertility Society (rAFS) stage III-IV endometriosis and 97 male blood donors in the UK, and (2) 130 women with rAFS stage III-IV endometriosis and 179 female newborn infants in Japan. No significant differences in allele or genotype frequencies were seen in either population. The genotype distribution in the UK population was 33/94 [35.1%] (cases) and 39/97 [40.2%] (controls) for A1A1 (homozygous wild-type); 43/94 [45.7%] (cases) and 44/97 [45.4%] (controls) for A1A2; and 18/94 [19.1%] (cases) and 14/97 [14.4%] (controls) for A2A2. The genotype distribution in the Japanese population was 31/130 [23.9%] (cases) and 57/179 [31.8%] (controls) for A1A1; 73/130 [56.2%] (cases) and 89/179 [49.7%] (controls) for A1A2; and 26/130 [20.0%] (cases) and 33/179 [18.4%] (controls) for A2A2. The CYP17 MspA1 polymorphism is probably not associated with endometriosis in either the UK or the Japanese population. PMID- 15823823 TI - Psychological effects of tibolone and sequential estrogen-progestogen therapy in perimenopausal women. AB - OBJECTIVE: To investigate changes in psychological symptoms before and after continuous tibolone treatment and sequential estrogen-progestogen therapy in perimenopausal women. METHODS: In this prospective, randomized, controlled study, perimenopausal women were randomly allocated to treatment with either tibolone 2.5 mg/day for 28 days (n = 28), or 0.625 mg conjugated equine estrogens (CEE) for 25 days plus 5 mg medroxyprogesterone acetate (MPA) daily on days 16-25 (n = 33). The differences in Beck's depression scores and serum lipid profiles before and after 1 year of treatment with both regimens were compared. RESULTS: Both groups were similar with respect to demographic characteristics. The differences in Beck's depression scores before and after treatment were statistically significant in the tibolone group (21.3 vs 17.1, p = 0.038) and also in the group receiving standard sequential estrogen-progestogen treatment (15.7 vs. 13.0, p = 0.040). In the sequential estrogen-progesterone group, a statistically significant increase was measured in high-density lipoprotein (HDL)-cholesterol levels after treatment (49.1 vs. 56.8 mg/dl, p = 0.023). CONCLUSION: Tibolone is as effective as sequential estrogen-progesterone therapy in alleviating the psychological symptoms of the perimenopause. In addition, CEE + MPA induces favorable changes in HDL-cholesterol. PMID- 15823824 TI - Serum adiponectin concentration and cardiovascular risk factors in climacteric women. AB - OBJECTIVE: Adiponectin plays a significant role in the modulation of glucose tolerance and insulin sensitivity. We attempted to evaluate the relationship between adiponectin level and parameters of the menopausal metabolic syndrome: body mass index, waist-to-hip ratio, lipid profile and insulin resistance indices. SUBJECTS AND METHODS: Thirty-two women and ten men aged 40-63 years were included. The percentage of body fat and of abdominal fat deposits were measured with dual-energy X-ray absorptiometry. Serum adiponectin, tumour necrosis factor a (TNFalpha) and leptin were measured with commercially available radioimmunoassay kits. To exclude the influence of nutritional factors on adiponectin secretion, diet content was analysed in the preceding three days. RESULTS: Postmenopausal non-obese women had a non-significantly lower level of adiponectin compared with premenopausal women of corresponding body mass. Serum adiponectin level was significantly lower in postmenopausal obese women than in non-obese women (p = 0.0023). Men with similar age and body mass to the women had the lowest level of adiponectin (p = 0.06). Three months of estrogen replacement therapy in women with surgical menopause did not significantly change the serum level of adiponectin. We found a negative correlation of adiponectin with leptin, insulin resistance index and total cholesterol, and a positive correlation with high-density lipoprotein cholesterol. Adiponectin level was negatively correlated with free testosterone, but we did not find such a relationship with estradiol. There was no correlation of adiponectin level with TNFalpha; however, serum TNFalpha correlated positively with leptin. The dietary analysis showed no differences between the diets of obese and non-obese women over the preceding three days. Moreover, mean diastolic and systolic blood pressures were noted to be significantly lower in premenopausal women than in postmenopausal non-obese women (p = 0.05). CONCLUSIONS: Our results suggest that adiponectin could be a marker of risk for developing menopausal metabolic syndrome. Moreover, it is possible that sex steroids have an influence on adiponectin secretion. PMID- 15823825 TI - Length of the menstrual cycle after discontinuation of oral contraceptives. AB - OBJECTIVE: To investigate whether the first cycle after stopping oral contraceptive (OC) intake had a normal duration. METHODS: A retrospective study was performed in 680 women, 300 non-OC users and 380 women discontinuing OC intake. The length of one or two menstrual cycles was recorded. RESULTS: In the non-user group, the median duration of both the first and second cycle was 29 days (range 18-97 and 20-56 days, respectively). In the OC user group the median duration from withdrawal bleeding until next menstruation was 30 (15-82) days. The second cycle lasted 29 (17-122) days. The duration of the first post treatment cycle was not significantly different from the next cycle or the cycle length in non-users. When the subjects were divided into different age categories, a significantly longer first post-treatment cycle was observed in the group aged 18-24 years, but a shorter first post-treatment cycle in the group aged 25-29 years. No differences were seen in the higher age groups. The ethinyl estradiol dose of the OC preparation did not influence the results. CONCLUSIONS: The first cycle after OC discontinuation had a normal duration. PMID- 15823826 TI - Effect of steroid add-back therapy on the proliferative activity of uterine leiomyoma cells under gonadotropin-releasing hormone agonist therapy. AB - Short-term treatment with gonadotropin-releasing hormone agonist (GnRHa) is a useful preoperative medical therapy of uterine leiomyomas. However, adverse effects caused by the hypo-estrogen state sometimes appear, suggesting the necessity of add-back therapy. In this study, we investigated effects of three kinds of add-back therapies on the proliferative activity of uterine leiomyoma cells by examining the expression of Ki-67 in leiomyoma cells by immunostaining. Thirty patients who were to undergo hysterectomy or myomectomy were injected with 3.75 mg depot leuprolide acetate every four weeks until the end of the 12th week. Twenty patients underwent add-back therapy from the 5th week to the end of the 12th week, 8 patients receiving 0.625 mg of conjugated equine estrogen (CEE) /day, 6 patients 5.0 mg of medroxyprogesterone acetate (MPA)/day, 6 patients 0.625 mg CEE plus 2.5 mg of MPA /day. The add-back of CEE or CEE plus MPA suppressed decreases in the proliferative activity of leiomyoma cells caused by GnRHa therapy, but that of MPA did not. These results suggest that the add-back therapy with MPA is of use in preventing the adverse effects caused by hypo estrogen in the preoperative short-term GnRHa therapy. PMID- 15823827 TI - Screening for insulin resistance in women with polycystic ovarian syndrome. AB - INTRODUCTION: Insulin resistance is implicated in the pathogenesis of polycystic ovarian syndrome (PCOS). Insulin-sensitizing agents are increasingly used in the treatment of infertility and hirsutism in PCOS. However, not all women with PCOS are insulin-resistant. OBJECTIVE: To assess the degree of insulin resistance within a clinic population of women referred for treatment of oligomenorrhoea or infertility. DESIGN: We evaluated 25 consecutive PCOS outpatients referred for treatment of menstrual dysfunction/infertility and a matched control group. All underwent a standard oral glucose tolerance test (OGTT) with serial insulin measurements. Insulin sensitivity was calculated using homeostasis model assessment (HOMA). RESULTS: Five of the 25 clinic patients had abnormal glucose handling (two had previously unknown type 2 diabetes and three had impaired glucose tolerance). Fasting and 2-h insulin levels were significantly higher in the PCOS women. Mean HOMA-S (insulin sensitivity) was even lower for PCOS women with normal GTT status (mean (95% confidence interval): 0.53 (0.34-0.72)) than for controls (0.94 (0.84-1.04)) (F = 4.2, p < 0.001). HOMA-B (pancreatic beta cell function) was nearly tripled for normal GTT status PCOS women at 273 (205 342) versus 105 (70-139) for controls (F = 6.8, p < 0.001). CONCLUSIONS: The results suggest a role for routine measurement of HOMA-S in identifying women with PCOS with insulin resistance with a view to targeting them with insulin sensitizing agents. PMID- 15823828 TI - High incidence of central precocious puberty in a bounded geographic area of northwest Tuscany: an estrogen disrupter epidemic? AB - The potential health consequences of human exposure to environmental estrogen disrupters are not known. Because many chemical compounds are environmentally persistent, toxic and estrogen-active, they can dysregulate the hypothalamic pituitary-gonadal axis, potentially inducing reproductive disorders such as central precocious puberty (CPP). We performed a multi-center analysis of CPP distribution in northwest Tuscany (NWT), an area of 5990 km2 with 1,280,895 inhabitants. Study criteria consisted of recorded CPP diagnoses and prescriptions of gonadotropin-releasing hormone analogs from January 1, 1998 to December 1, 2003. Although similar CPP prevalences were found in four major cities of NWT (Livorno, Lucca, Massa and Pisa) (mean 30.4 per 100,000 children, standard deviation 18.6; p > 0.05), Viareggio area (< 300 km2) with 19,219 child inhabitants (0-14 years of age) had the highest CPP prevalence: more than 161 CPP cases per 100,000 children. Living in Viareggio area significantly increased the risk of CPP (relative risk (RR) 5.73, 95% confidence interval (CI) 3.5-9.3; rate/risk difference 0.133%, p < 0.05). Annual CPP incidence in the Viareggio area was relatively constant and significantly higher than in other NWT areas (RR 5.04, 95% CI 2.3-11.2; rate/risk difference 0.03%, p < 0.05). Indeed, 47% of total NWT cases were distributed in the countryside (300?km2) surrounding Viareggio. Specifically, three villages - Camaiore, Pietrasanta and Stazzema - in Viareggio presented the highest CPP frequency: 216.1, 393.5 and 274.0 CPP cases per 100,000 children, respectively (RR 9.59, 95% CI 1.71-16.6; rate/risk difference 0.26%, p < 0.05). Owing to the definite geographic distribution of CPP and because increasing distance (km) from Pietrasanta rarefied CPP frequency, we suggest environmental factors (e.g. estrogen disrupter pollution) as major CPP determinants in NWT. PMID- 15823829 TI - Sexual hormone serum levels and temporomandibular disorders. A preliminary study. AB - The aim of the present study was to investigate the role of sexual hormones in a young adult population affected by articular forms of temporomandibular disorders (TMD), measuring 17beta-estradiol and progesterone serum levels. In the study, we included 40 patients (20 males and 20 females) with a Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) axis I group II diagnosis of disk displacement and/or group III diagnosis of arthralgia, osteoarthritis or osteoarhrosis, and 32 healthy controls. In female patients, blood samples were collected in follicular and luteal phases of the same menstrual cycle, while only one blood sample was drawn in male patients. Serum levels of estradiol and progesterone were determined using a radioimmunoassay and the comparison between the two groups was performed using a t test. Regarding estradiol, our results showed significantly higher serum levels in patients affected by TMD than in healthy controls, both in males (p < 0.01) and in the luteal phase of the menstrual cycle in females (p < 0.05), while no difference was found for progesterone serum levels. Considering the multifactorial etiology of TMD and the hypothesis that some joint tissues (e.g., bone, cartilage, collagen, proteins) could be a target for sexual hormones, these data suggest that high serum estrogen levels might be implicated in the physiopathology of TMD. PMID- 15823830 TI - The effect of relcovaptan (SR 49059), an orally active vasopressin V1a receptor antagonist, on uterine contractions in preterm labor. AB - Relcovaptan (SR 49059) is a non-peptide, orally active vasopressin V1a receptor inhibitor. The effect on uterine contractions in 18 women with preterm labor in pregnancy weeks 32-36 was assessed in a double-blind investigation. The inclusion criterion was at least four regular uterine contractions over 30 min as measured by external tocodynamometry. Twelve patients received at random a single oral dose of 400 mg relcovaptan and six received placebo, and contractions were monitored up to 6 h thereafter. Rescue medication (beta-adrenoceptor-stimulating drug) was allowed after 2 h. Before drug administration a mean (+/- SE) of 8.2 +/ 1.4 and 9.7 +/- 1.6 contractions/30 min were recorded in the relcovaptan- and placebo-treated groups, respectively. In the former group, the frequency of uterine contractions started to decrease within the first half hour, and 1.5-2 h after dosing it was steady at 3.2 +/- 0.9 contractions/30 min. Correspondingly, after placebo, 7.8 +/- 2.2 contractions/30 min were recorded, a statistically significant difference (p = 0.017). The activity in the relcovaptan-treated women remained low, whereas in the placebo group inhibited uterine contractions were observed only in women receiving 'rescue' tocolytic treatment. It is concluded that relcovaptan inhibits preterm labor. PMID- 15823831 TI - Different effects of tibolone and continuous combined estrogen plus progestogen hormone therapy on sex hormone binding globulin and free testosterone levels--an association with mammographic density. AB - OBJECTIVE: To compare the effects of tibolone and continuous combined hormone therapy on circulating sex steroids and their binding proteins and their relationship to mammographic density. STUDY DESIGN: A prospective, double-blind placebo-controlled study. A total of 166 postmenopausal women were equally randomized to receive tibolone 2.5 mg, estradiol 2 mg/norethisterone acetate 1 mg (E2/NETA) or placebo. Serum analyses of sex steroids, insulin-like growth factor (IGF-I) and binding proteins and assessment of mammographic breast density were performed at baseline and after 6 months of treatment. RESULTS: Estrogens were markedly increased and androgens decreased by E2/NETA. In contrast, tibolone had only a minor influence on circulating estrogens. Sex hormone binding globulin (SHBG) levels were reduced by 50%, while levels of androgens increased. Baseline values of estrone sulfate (E1S), around 1.0-1.1 nmol/l, were increased to 44.7 nmol/l by E2/NETA and to only 1.7 nmol/l by tibolone (p < 0.001). Mammographic breast density displayed a negative correlation with age and body mass index and a positive association with SHBG. After 6 months there was also a negative correlation with levels of free testosterone. Conclusion We found that tibolone and E2/NETA caused distinct differences in estrogen/androgen status and blood levels of possible breast mitogens. The negative association between free testosterone and mammographic density could be a possible explanation for tibolone having less influence on the breast. PMID- 15823832 TI - Hormone therapy and cardiovascular disease--still much to be learnt. AB - Results from the recent randomized clinical trials indicating that hormone therapy (HT) does not provide cardiovascular protection, but potential harm, are in profound disagreement with the sound evidence from numerous observational and experimental studies. While the observational studies have mainly assessed symptomatic recently menopausal women, the randomized trials have studied symptomless elderly postmenopausal women with established coronary heart disease or various risk factors for cardiovascular disease. Thus, the recent trials have revealed only that HT does not provide secondary cardiovascular benefits. Since primary cardiovascular benefits of HT are rational but not yet proven in clinical trials, new studies are in demand. Until more data from recently menopausal symptomatic women are available, we need to base our decisions on existing evidence and good clinical practice. Although the potential of HT to provide cardiovascular benefits is decreased by advancing age and time since menopause, this should not preclude the use of individualized HT in younger postmenopausal women. PMID- 15823844 TI - National Occupational Standards in clinical embryology: implementation in 2005? AB - In 2001 the UK Department of Health initiated the National Occupational Standards Project in Healthcare Science. This project incorporates the 43 disciplines of healthcare science with the aim of developing a framework for competent performance in all areas of healthcare science, thereby standardising the delivery of services in this area. The whole of the project relies on the concept of function, and using the process of functional analysis each discipline is broken down into key work areas. The standards are derived from a further breakdown of these key work areas. Each standard gives expected requirements for the competent performance of the function, along with the relevant knowledge required. It is envisaged that the implementation of National Occupational Standards will ensure that individuals performing healthcare science functions throughout the UK, whether they are healthcare scientists themselves or not, will be performing these functions to an adequate standard. Since work began on the project 63 standards, and associated assessment guidance, have been drafted the whole of healthcare science. The National Occupational Standards are due to be implemented in 2005 by Skills for Health, an organisation that defines and monitors skills in healthcare both in the NHS and the private sector. PMID- 15823845 TI - Lives to save lives--the ethics of tissue typing. AB - Should we allow tissue typing of in vitro embryos in order to implant those which could provide potentially life-saving cells to an existing serious ill sibling with that tissue type? A case is made that such tissue matching does not involve unacceptable instrumentality towards or commodification of children. The key distinction is that the parents' request for tissue typing is reactive in the face of serious medical need rather than being proactive in the sense of seeking the means to specify a child with chosen desirable characteristics. Nevertheless, as preimplantation genetic diagnosis (PGD) is a relatively new technique, both long-term safety issues concerning effects on child development following embryo biopsy and the risks of misdiagnosis must be given due weight as must the avoidance of exploitation of couples desperate to save a sick child. The HFEA originally made a distinction, recently revoked, between allowing tissue typing after PGD to select against affected embryos and denying it when PGD is not required because the embryos are not at risk of inheriting the disease suffered by the existing sibling. If tissue typing is not inherently unethical and misdiagnosis poses a greater risk than biopsy damage, then this distinction is not ethically tenable. PMID- 15823846 TI - The welfare of the child. PMID- 15823847 TI - The genetic and cytogenetic basis of male infertility. AB - Despite the difficulties in determining the relative maternal vs. paternal contributions to infertility it is often suggested that a male factor problem is implicated in 50% of cases. This review is concerned specifically with male fertility disorders that have a clearly defined genetic component. The genetic causes of infertility can be broken down into Y chromosome deletions (specifically deletions in the AZF a, b, and c regions), single gene disorders (particularly those relating to the CFTR gene), multifactorial causes and chromosome abnormalities. Chromosome abnormalities can be numerical (such as trisomy--full blown or mosaic) or structural (such as inversions or translocations). Of especial interest at present is the incidence of levels of numerical chromosome abnormalities in the sperm of infertile men; prospects for screening sperm for such abnormalities are discussed. PMID- 15823848 TI - Myomectomy: a review of surgical technique. AB - Fibroids are common amongst women of reproductive age. In women wanting to retain their fertility and/or uterus open myomectomy remains the most widely performed procedure. It is important to carefully counsel the patient about the pros and cons of surgery versus expectant management. Meticulous pre-operative evaluation and preparation will help ensure the procedure goes smoothly. Attention to the operating technique used is paramount in ensuring patient safety and surgical peace of mind. This review addresses the important aspects of open myomectomy including operative technique. PMID- 15823849 TI - Womb with a view. AB - Zygote transfer is a significant limiting factor in pregnancy rates after Assisted Reproductive Technology (ART) treatment. Adequate technique appraisal should be a high priority task for any medical facility that offers this treatment. Although many clinicians have extensive personal experience, it is difficult for trainees to understand the anatomical and mechanical process of safe and adequate placing of zygotes in the uterine cavity. Ultimately, even with ultrasound help, it is a "blind procedure". The experimental module described here allows for direct visualisation of the mechanical passage of a catheter through the cervix and final placement of the tip in relation to the intravaginal visible part of the catheter. It has worldwide practical applicability for trainees and experienced clinicians accompanying the theory and allowing adequate training and retraining of all staff performing zygote transfers. PMID- 15823853 TI - Quantification of dung carbon incorporation in a temperate grassland soil following spring application using bulk stable carbon isotope determinations. AB - Herbivore dung constitutes a substantial input of C to temperate grassland soils, and its fate must be determined in order to fully understand nutrient cycling in this ecosystem. This experiment used changes in bulk delta13C values of the 0-1 cm and 1-5 cm soil horizons of a dung-treated temperate grassland soil to approximate percentage applied dung C incorporation over 372 days. Natural abundance 13C-labelled C4 dung (delta13C - 12.6%) and C3 dung (delta13C - 31.3% were produced in a monitored diet switch from ryegrass silage (delta13C - 30.1%) to maize silage (delta13C - 11.6%). The dung was applied to a C3 grassland (delta13C 0-1 cm - 29.9%, 1-5 cm - 30.6%), and dung remains and soil cores from beneath the treatments were sampled at intervals. delta13C values were used to estimate a maximum of 12% applied dung C incorporation in the top 5 cm of the soil after 112 days, which declined to around 8% at the end of the experiment. A significant increase in percentage applied dung C was observed in the top 1 cm of soil, compared with the 1-5 cm horizon, after a substantial rain event after 30 days. However, results of forage fibre analyses of the two dung types revealed significant differences in composition which may affect subsequent calculations of percentage dung incorporation based on bulk delta13C values. PMID- 15823854 TI - Using 15N to determine a budget for effluent-derived nitrogen applied to forest. AB - Using stable N isotopes, the fate of effluent-derived N has been determined within a land based municipal effluent irrigation scheme. Over 900 metric tonnes(t) of effluent-derived N have been applied to 192 ha of production conifer forest near Rotorua (NZ) over the past 11 years. The effluent N has a natural isotopic signal, generated by the treatment process, allowing it to be traced into various components of the system. Using this isotopic signal, a realistic approximation of storage capacity of various components of the system has been generated, including a calculation of the contribution of effluent N exiting the catchment via stream flow. Forest storage accounts for 50% of the applied N with a considerable proportion of that immobilized in wood and soil. The wetland, although not intensively sampled, retains 115 t, (13%) of the applied N. Denitrification, including that occurring within the wetland, accounts for 23 t (3%). Nitrogen isotope data confirm that the rise in NO3 concentrations is directly attributable to effluent N. Currently 88% of NO3-N in the stream is effluent-derived. Using current N isotope values for the stream and extrapolating over the discharge period, export of effluent N via the stream is estimated as 263 t (29%) of the applied N. Overall the forest and wetland ecosystem has intercepted or denitrified 65% of applied N, with 29% lost to the stream, and 50 t (5%) unaccounted for. The forest ecosystem is currently over-supplied with N and a number of management implications flows from these findings. In the long term the continued application of effluent N to the current irrigation area is not sustainable. PMID- 15823855 TI - Using natural 15N abundances to trace the fate of waste-derived nitrogen in forest ecosystems: New Zealand case studies. AB - Treatment of wastewater generally results in elevated natural 15N abundance (delta15N) in the effluent and sludges. For example, high delta15N values are found in treated sewage effluent, biosolids, and other wastes that are commonly applied to land. In contrast, N deficient coniferous forest soils usually have a low delta15N. When wastes with high delta15N values are applied to land, their distinctive delta15N signature can potentially be used to trace the fate of waste derived N in the ecosystem. In this paper, we provide an overview of the use of delta15N in land application of wastes, including New Zealand case studies on tracing nitrogen in forest ecosystems. PMID- 15823856 TI - Isotope studies to the sorption behavior of atmospheric sulfate in humus layers of scots pine ecosystems. AB - The sorption potential for SO4(2-) in humus layer samples from field sites along a deposition gradient was determined experimentally in batch experiments. The Freundlich equation was used to quantify the sorption of added SO4(2-) in humus layer samples and to determine site-dependent sorption parameters. SO4(2-) sorption in humus layers is a concentration-dependent process. The linearity of isotherms reveals that SO4(2-) is reversibly bound in the organic surface layer, as long as soil solution concentrations remain above 26 to 44 mg SO4(2-) L(-1). Natural isotope variations of sulfur in SO4(2-) were analysed to investigate the degree of sorption of dissolved atmospheric and added SO4(2-). Both sulfate species differed significantly in their isotope composition. The pattern of delta34S values for SO4(2-) in all equilibrium solutions confirm the findings from sorption isotherms, showing a close relationship between the sulfur isotope ratios of SO4(2-) in soil solutions and the amount of SO4(2-) sorbed at the humus layer matrix. Stored atmospheric SO4(2-) in humus layers is released at sites where sulfate concentration in throughfall drops below 26 mg SO4(2-) L(-1). Concentration of soluble Fe decreased with increasing sulfate sorption, thus supporting the assumption that active Fe for example is important. Iron probably stabilizes the reactive surface of humus complexes and therefore has a positive influence on the SO4(2-) sorption in humus layers. PMID- 15823857 TI - A groundwater separation study in boreal wetland terrain: the WATFLOOD hydrological model compared with stable isotope tracers. AB - Monitoring of stable water isotopes (18O and 2H) in precipitation and surface waters in the Mackenzie River basin of northern Canada has created new opportunities for researchers to study the complex hydrology and hydroclimatology of this remote region. A number of prior studies have used stable isotope data to investigate aspects of the hydrological regime of the wetland-dominated terrain near Fort Simpson, Northwest Territories, Canada. The present paper compares estimates of groundwater contributions to streamflow derived using the WATFLOOD distributed hydrological model, equipped with a new water isotope tracer module, with the results of conventional isotope hydrograph separation for five wetland dominated catchments along the lower Liard River. The comparison reveals highly promising agreement, verifying that the hydrological model is simulating groundwater flow contributions to total streamflow with reasonable fidelity, especially during the crucial snowmelt period. Sensitivity analysis of the WATFLOOD simulations also reveals intriguing features about runoff generation from channelized fens, which may contribute less to streamflow than previously thought. PMID- 15823858 TI - Regional variation and relationships between the contaminants dde and selenium and stable isotopes in swallows nesting along the Rio Grande and one reference site, Texas, USA. AB - Cave swallows (Petrochelidon fulva) and cliff swallows (P. pyrrhonota) nest in numerous colonies throughout the Texas portion of the Rio Grande along the U.S. border with Mexico. We collected swallows during 1999 and 2000 from eight locations along the Rio Grande to determine if delta15N and delta13C values could be used to predict 1,1-di-(p-chlorophenyl-)2,2-dichloroethene (DDE) and selenium (Se) contaminant burdens in insectivorous birds nesting across a geographic gradient in the Texas-Mexico border and to discern if stable isotopes could help discriminate between local versus nonlocal acquisition of contaminants. We analysed delta15N and delta13C in liver and muscle and DDE and Se in swallow carcasses. Within individuals, delta15N was higher in liver than in muscle of both species by an average of 1.34%, whereas delta13C was 0.145% higher in muscle than in liver. Significant differences occurred among locations in delta15N and delta13C values in liver and muscle of both species. Cave swallows from three locations in the Lower Rio Grande Valley were more enriched in delta15N than swallows from other sites. In general, swallows nesting in more northern latitudes along the Rio Grande had lower delta15N and delta13C values than those nesting farther south. Concentrations of DDE were significantly greater in swallows from El Paso, Llano Grande, and Pharr than in those from Brownsville, Falcon Lake, Laredo, Del Rio, and a reference site outside the Rio Grande. All swallows (n = 21) from El Paso, Llano Grande, and Pharr had DDE concentrations > or = 3 microg g(-1) wet weight (ww), a value three times greater than the estimated threshold in avian prey that could cause potential reproductive failures in raptors. Concentrations of Se also were significantly greater in El Paso and Del Rio than at other locations. Most Se concentrations were not of concern for direct effects on birds or their predators. Principal component analysis indicated some positive correlations between delta15N and delta13C values in tissues and contaminant concentrations in carcass; however, analysis of covariance suggested a stronger effect of location on concentrations of DDE and Se. At the local level (Llano Grande and Pharr) there was a significant positive correlation between delta15N in liver and DDE concentrations in swallow carcasses; however, Se concentrations were not significantly correlated with isotopes even at the local level. Our results provide a good database of delta15N and delta13C values for insectivorous birds nesting along the Rio Grande. Similar ecologies for cave and cliff swallows and their abundance and wide distribution along the Rio Grande make them ideal indicators of environmental pollution of this portion of the Texas-Mexico border. PMID- 15823859 TI - Temporal diet changes recorded by stable isotopes in Asiatic black bear (Ursus thibetanus) hair. AB - Carbon and nitrogen stable isotope ratios were measured in hair samples of the Asiatic black bear (Ursus thibetanus) inhabiting the Northern Japanese Alps (NJA) (n = 20) and the periphery of Nagano City (NC) (n = 6), in Nagano Prefecture, Japan. The hair of NJA bears, which did not have access to anthropogenic foods, showed lower values of d13C and d15N than that of NC bears which had access to garbage and corn fields, especially during the summer. These results reflect somewhat differing diets between the NJA and NC bears. We attempted to assess the feeding history during the hair growth cycle using the growth section analysis method. Each hair sample had been cut into 3?mm lengths from root to tip, labeled, and analyzed along the hair growth. We measured the carbon and nitrogen stable isotope ratios of each 3?mm length of hair sample from one NC bear which had been killed while raiding a corn field. The sections showed wide ranges of isotope ratios, from -23.2% to -14.6% for delta13C, and from 0.3% to 4.6% for delta15N. It was shown that the diet of this bear shifted dramatically from principally C3 plants to more C4 plants and to foods of animal origin. An analysis of the whole hair reflects just the average feeding habit during hair growth, but the present method can trace its diet history. This method can contribute to obtain precise ecological information of wildlife. PMID- 15823861 TI - Effect of statins and aspirin alone and in combination on clinical outcome in dyslipidaemic patients with coronary heart disease. A subgroup analysis of the GREACE study. AB - DECLARATION OF INTEREST: The GREACE study was conducted independently; no Company or Institution has supported it financially. Some of the authors have attended conferences and participated in other trials sponsored by various pharmaceutical companies. We assessed the possible 'synergy' of statins and aspirin (ASA) in reducing vascular events in patients with coronary heart disease, in a post hoc analysis of the GREek Atorvastatin and Coronary-heart-disease Evaluation (GREACE) study. All patients (n = 1600) were divided into four groups according to long term treatment: Group A (n = 787; statin + ASA), B (n = 93; statin - no ASA), C (n = 599; no statin - on ASA) and D (n = 121; no statin - no ASA). From all patients 692 were either on a statin or ASA monotherapy (Groups B + C). Relative risk reductions (RRRs) in 'all events' (primary endpoint) between groups were assessed. During the 3-year follow-up there were 292 cardiovascular events; 92 (12% of patients) in Group A, 14 (15%) in group B, 144 in Group C (24%) and 42 events in Group D (35%). The total number of events in Group B + C was 158 (23%). The RRRs in the primary endpoint were: Group A versus B 24% (P = 0.1912), A versus C 51% (P < 0.0001), A versus B + C 49% (P < 0.0001) and A versus D 71% (P < 0.0001). The RRRs in Group B versus C was 36% (P = 0.0431) and B versus D 57% (P = 0.0012), while in C versus D 33% (P = 0.0084). Our findings show that statins and ASA have an additive effect in reducing cardiovascular events. Aggressive statin use in the absence of ASA also substantially reduced cardiovascular events. Treatment with ASA in the absence of statin use reduced clinical events in comparison to patients not treated with either drug. PMID- 15823863 TI - Platelet activation can occur by shear stress alone in the PFA-100 platelet analyser. AB - The PFA-100 platelet function analyser simulates high-shear platelet function; however, the relative importance of the high shear rates alone has not been examined in this system. Normal blood was analysed by the PFA-100 using modified cartridges that contained a blank membrane lacking the usual platelet agonists (collagen/epinephrine or collagen/ADP) thus isolating the effects of shear stress alone. We found evidence of platelet activation with significant increases in platelet-leucocyte aggregates. There was no significant change in surface CD62P nor PAC-1 expression. A degree of platelet activation occurs in the PFA-100 due to shear stress alone. PMID- 15823862 TI - Inhibitory effects of P2Y12 receptor antagonists on TRAP-induced platelet aggregation, procoagulant activity, microparticle formation and intracellular calcium responses in patients with acute coronary syndromes. AB - Thrombin induces platelet aggregation and membrane rearrangements leading to enhanced procoagulant activity and microparticle production, all of which are thought to contribute to thrombus formation in patients with acute coronary syndromes (ACS). Clopidogrel, an adenosine diphosphate (ADP) receptor antagonist acting at the P2Y(12) receptor, has been shown to provide clinical benefit in ACS. We aimed to investigate the effects of clopidogrel ex vivo and another ADP antagonist, AR-C69931MX in vitro on thrombin receptor activating peptide (TRAP) induced platelet aggregation, procoagulant activity, microparticle formation and [Ca(2+)]i responses in patients with ACS. Measurements were performed in platelet rich plasma using aggregometry and flow cytometry (n = 12). Clopidogrel (300 mg loading dose plus 75 mg daily) significantly inhibited TRAP-induced aggregation, procoagulant activity (annexin V binding) and microparticle production (all P < 0.05) but not as extensively as AR-C69931MX (400 nmol/l). [Ca(2+)]i responses induced by a combination of TRAP and ADP designed to mimic the physiological effects of released ADP showed that clopidogrel partially and AR-C69931MX completely removed the ADP component of the [Ca(2+)]i responses (n = 6). The results provide new information on the mechanisms involved in the beneficial effects of P2Y(12) antagonists in patients with ACS. PMID- 15823864 TI - Co-existence of Bernard Soulier syndrome and factor XI deficiency in a family: a unified pathology? AB - Bernard Soulier syndrome (BSS) is an autosomal recessive disorder of platelet function. Factor XI deficiency leads to a variable bleeding tendency and the defect is also inherited in an autosomal recessive manner. In this paper we describe a case of BSS with severe deficiency of factor XI. The patient had both BSS and factor XI deficiency. The sister and both the parents were heterozygous for BSS and had factor XI deficiency. The brother is normal for both BSS and factor XI levels. Apart from the patient, no other family members had any history of bleeding in spite of having a deficiency of factor XI, which suggests that low level of factor XI in this family was not responsible for bleeding. Curiously, although the index patient inherited both Bernard Soulier syndrome and factor XI deficiency (FXI: C = 1.3%), he had mild bleeding symptoms restricted only to ecchymoses and petechiae. Detailed review of the pedigree showed that all the members who inherited the BSS phenotype, also inherited abnormal factor XI gene. Karyotype of the affected members of the family using standard Giemsa banding technique showed a normal picture. Considering the fact that the genes causing both BSS and factor XI are both on widely different chromosomes, their coinheritance in four members of the family without any unusual translocation suggest a unified pathology probably on the basis of a common transcription factor defect or a common post translational processing defect. This is the first case of coinheritance of BSS and factor XI deficiency reported in the English literature. PMID- 15823865 TI - Leukocyte-platelet interaction in pregnancies complicated with preeclampsia. AB - OBJECTIVE: Pregnancy is characterized by haemostasis activation, and in preeclampsia endothelial dysfunction, platelet and leukocyte activation are further characteristic features. The aim of this study was to investigate to which extent platelets from normotensive pregnant women or those with preeclampsia are circulating as microparticles or platelet-platelet aggregates. We also investigated if platelet-leukocyte multiconjugates were differently present in nonpregnant and pregnant women. STUDY DESIGN: Using flow cytometry we investigated these parameters in basal samples and after in vitro stimulation with adenosine diphosphate (ADP) or thrombin receptor activation peptide. This was done in samples from 20 matched preeclamptic and normotensive pregnant women and in a group of 12 nonpregnant women. RESULTS: In the basal state we found that women with preeclampsia had a smaller portion of microparticles circulating than the normotensive pregnant women, Upon ADP stimulation both pregnancy groups showed a higher percentage of monocytes and granulocytes with platelets attached and also a higher number of platelets attached to each monocyte and granulocyte than in the group of nonpregnant individuals. CONCLUSION: This article presents further evidence that changes from the nonpregnant to the pregnant state are associated with hemostasis activation as an integrated part of an inflammatory reaction that is even more pronounced when pregnancy is complicated with preeclampsia. PMID- 15823866 TI - Expression of a recombinant protein of the platelet F11 receptor (F11R) (JAM 1/JAM-A) in insect cells: F11R is naturally phosphorylated in the extracellular domain. AB - The F11 receptor (F11R/JAM) is a member of the immunoglobulin superfamily localized on the membrane surface of human platelets and a component of tight junctions of endothelial and epithelial cells. F11R was demonstrated to participate in the adhesion of human platelets to cytokine-inflamed endothelial cells (EC), indicating an important role for F11R in inflammatory thrombosis and atherosclerosis. Domains responsible for the formation of tight junctions, the adhesion of platelets to EC, activation of platelets resulting in granule release, the activation of IIb/3 integrin and platelet aggregation, were identified in the external portion of F11R. To further examine critical sites of F11R, we utilized the baculovirus system to generate the F11R recombinant protein with the sequence of the extracellular domain, in two types of insect cells, Sf9 and H5. The F11R recombinant protein was detected in the cytoplasm of both infected Sf9 and H5 insect cells, but only infected H5 cells secreted a soluble F11R protein. The purified recombinant F11R proteins, obtained from both types of insect cells, were recognizeable by a conformation-dependent monoclonal antibody, M.Ab.F11, directed against domains within the N-terminus and the first Ig-like fold of F11R. Assessment of the phosphorylation state in the recombinant F11R protein revealed phosphorylation of serine, threonine and tyrosine amino acid residues within the external domain. Real-time biomolecular interaction analysis, performed to assess kinetic constants associated with the binding of active molecules to the purified recombinant F11R protein revealed high affinity binding of the phosphorylated recombinant protein by M.Ab.F11 with K(a) of 5.47 x 10(6) and K(d) of 1.83 x 10(-7), comparable to values measured with intact human platelets. The findings reported here provide new information on specific domains of F11R that can lead to the generation of therapeutic agents expected to be useful in the treatment of cardiovascular diseases. PMID- 15823867 TI - Insensitivity to the alpha2-adrenergic receptor blocker yohimbine hydrochloride and occurrence of spontaneous platelet macroaggregation (SPMA) in diabetes. AB - We report here a study of platelet aggregation in diabetes, induced by epinephrine and its inhibition by yohimbine hydrochloride (YH), an alpha(2) adrenergic receptor-blocking agent. Interestingly, emergence of spontaneous platelet macroaggregation (SPMA) was observed in six out of 75 cases in the absence of any agonist. The SPMA cases were strongly associated with insensitivity to YH (in contrast with non-SPMA cases) when epinephrine was used as an agonist. We suggest that the observed correlation is a result of over expression of platelet alpha(2)-adrenoceptors in such subjects. The quantitative nature of the effect is supported by the observation that addition of YH at higher concentration (more than 5 microM) led to restoration of the adrenergic receptor-blocking activity of the said agent. Eventually for non-SPMA subjects YH exhibited blocking activity even at lower concentration. The aggregation profile and the platelet morphology of the SPMA cases had distinctive features as compared to microaggregates formed in other diabetic subjects (non-SPMA cases). PMID- 15823868 TI - High prevalence of sustained remission of idiopathic thrombocytopenic purpura after Helicobacter pylori eradication: a long-term follow-up study. AB - We studied the effect of Helicobacter pylori (H. pylori) eradication in 43 consecutive H. pylori-infected patients with idiopathic thrombocytopenic purpura. H. pylori was eradicated with antibiotics in 41 of them (95.3%). The difference between the mean platelet count before and after therapy was statistically significant (54.3 x 10(9)/l vs. 119.1 x 10(9)/l; P < 0.001). A sustained remission was observed in 20 patients (48.8%), after a median follow-up of 31.2 months. None of the patients still infected by H. pylori after therapy reached normal platelet values. The long-term follow-up confirms the efficacy of H. pylori eradication in H. pylori-infected ITP patients. PMID- 15823869 TI - Platelets are covercytes, not phagocytes: uptake of bacteria involves channels of the open canalicular system. AB - Recent studies have suggested that uptake of bacteria by human platelets is more similar to ingestion of the same organisms by neutrophils than previously considered. Platelet pseudopods surround a bacterium by a process of circumferential adherence until the organism is fully enclosed in a vacuole consisting of internalized extracellular space surrounded by plasma membrane. The surface connected, open canalicular system (OCS) is not involved in the process of platelet phagocytosis. The present investigation has reviewed the literature on platelet-particle and platelet-bacterial interaction, and added a new study of the process of bacterial engulfment employing tannic acid as an electron dense tracer. The osmium black reaction product formed by tannic and osmic acids during fixation stained the glycocalyx on the platelet exterior surface and lining channels of the OCS. It also stained the vacuoles containing bacteria in platelets as well as channels of the OCS connected to the engulfment vacuole and exterior surface of the cell. Alpha granules releasing their contents into the OCS and into engulfment vacuoles were also stained by osmium black. The only way that tannic acid could reach and stain the interior surface of engulfment vacuoles during fixation was through patent channels of the OCS. In contrast, neutrophils do not contain an OCS, and phagocytic vacuoles containing organisms are never stained by tannic acid. Thus the platelet is a covercyte, not a phagocyte, and uptake of the bacteria does involve channels of the OCS. PMID- 15823870 TI - The field performance of frontal air bags: a review of the literature. AB - This article presents a broad review of the literature on frontal air bag field performance, starting with the initial government and industry projections of effectiveness and concluding with the most recent assessments of depowered systems. This review includes as many relevant metrics as practicable, interprets the findings, and provides references so the interested reader can further evaluate the limitations, confounders, and utility of each metric. The evaluations presented here range from the very specific (individual case studies) to the general (statistical analyses of large databases). The metrics used to evaluate air bag performance include fatality reduction or increase; serious, moderate, and minor injury reduction or increase; harm reduction or increase; and cost analyses, including insurance costs and the cost of life years saved for various air bag systems and design philosophies. The review begins with the benefits of air bags. Fatality and injury reductions attributable to the air bag are presented. Next, the negative consequences of air bag deployment are described. Injuries to adults and children and the current trends in air bag injury rates are discussed, as are the few documented instances of inadvertent deployments or non-deployment in severe crashes. In the third section, an attempt is made to quantify the influence of the many confounding factors that affect air bag performance. The negative and positive characteristics of air bags are then put into perspective within the context of societal costs and benefits. Finally, some special topics, including risk homeostasis and the performance of face bags, are discussed. PMID- 15823871 TI - Driver mortality in frontal crashes: comparison of newer and older airbag designs. AB - In 1997, the National Highway Traffic Safety Administration amended its requirements for frontal crash performance under Federal Motor Vehicle Safety Standard 208 to temporarily allow 30 mi/h (48 km/h) sled tests with unbelted dummies as an alternative to 30 mi/h head-on rigid-barrier vehicle tests. This change permitted automakers to reduce airbag inflation forces so that they would be less likely to injure occupants who are close to airbags when they first deploy. Most vehicle models were sled-certified starting in model year 1998. Airbag-related deaths have decreased since 1997; however, controversy persists about whether reduced inflation forces might be decreasing protection for some occupants in high-severity frontal crashes. To examine the effects of the regulatory changes, this study computed rate ratios (RR) and 95% confidence intervals (95% CI) for passenger vehicle driver deaths per vehicle registration during 2000-2002 at principal impact points of 12 o'clock for 1998-99 model year vehicles relative to 1997 models. Passenger vehicles included in the study had both driver and passenger front airbags, had the same essential designs during the 1997-1999 model years, and had been sled-certified for drivers throughout model years 1998 and 1999. An adjustment was made for the higher annual mileage of newer vehicles. Findings were that the effect of the regulatory change varied by vehicle type. For cars, sport utility vehicles, and minivans combined, there was an 11 percent decrease in fatality risk in frontal crashes after changing to sled certification (RR = 0.89; 95% CI = 0.82-0.96). Among pickups, however, estimated frontal fatality risk increased 35 percent (RR = 1.35; 95% CI = 1.12 1.62). For a broad range of frontal crashes (11, 12, and 1 o'clock combined), the results indicated a modest net benefit of the regulatory change across all vehicle types and driver characteristics. However, the contrary finding for pickups needs to be researched further. PMID- 15823872 TI - Relationships of frontal offset crash test results to real-world driver fatality rates. AB - In a study of the relationship between Insurance Institute for Highway Safety frontal offset crash test ratings and real-world fatality rates, there was a clear trend for better-rated vehicles to have lower driver fatality risk, although the correlation was not uniform across all vehicle groups or statistically significant in all cases. For all types of crashes combined, fatality rates per registered vehicle were generally lower for vehicles rated good than for vehicles rated poor, but rates for acceptable and marginal vehicles were not always within this range. A more precise examination of fatality risk was accomplished by comparing driver outcomes in fatal two-vehicle crashes. When a rated vehicle collided with a nonrated vehicle, the fatality risk for the rated vehicle driver was highest for poorly rated vehicles, then progressively smaller for vehicles with marginal, acceptable, or good ratings. For two-vehicle crashes of similar vehicles rated good and poor, the odds of driver fatality was 34 percent lower for the good vehicle than for the poor vehicle, but this estimate was not statistically significant. Finally, in head-on crashes of rated vehicles, the estimated odds of driver fatality was approximately 74 percent lower for the good vehicle than for the poor vehicle, with confidence limits ranging from 28 to 91 percent. PMID- 15823873 TI - Variations in occupant response with seat belt slack and anchor location during moderate frontal impacts. AB - Both seat belt slack and anchor location are known to affect occupant excursion during high-speed frontal collisions, but their effects have not been studied at moderate collision severities. The goal of this study was to quantify how seat belt slack and anchor location affect occupant kinematics and kinetics in moderate severity frontal collisions. A Hybrid III 50th percentile male dummy was seated on a programmable sled and exposed to frontal collisions with a speed change of 17.5 km/h. The seat belt was adjusted either snugly or with 10 cm slack (distributed 60/40 between the shoulder and lap portions) and the anchor location was varied by adjusting the seat position either fully forward or rearward (seat travel = 13 cm). Accelerations and displacements of the head, T1 and pelvis were measured in the sagittal plane. Upper neck loads and knee displacements were also measured. Five trials were performed for each of the four combinations of belt adjustment (snug, slack) and anchor location (seat forward, seat rearward). For each trial, kinematic and kinetic response peaks were determined and then compared across conditions using ANOVAs. Peak displacements, accelerations and loads varied significantly with both seat belt slack and anchor location. Seat belt slack affected more parameters and had a larger effect than anchor location on most peak response parameters. Head displacements increased a similar amount between the snug/slack belt conditions and the rearward/forward anchor locations. Overall, horizontal head displacements increased from 23.8 cm in the snug-belt, rearward-anchor configuration to 33.9 cm in the slack-belt, forward-anchor configuration. These results demonstrated that analyses of occupant displacements, accelerations and loads during moderate frontal impacts should consider potential sources of seat belt slack and account for differences in seat belt anchor locations. PMID- 15823874 TI - Drug and drink driving by university students: an exploration of the influence of attitudes. AB - The present study aimed to gather information on drug and alcohol use, the prevalence of drink and drug driving and attitudes to both in an undergraduate university cohort (n = 275). Further, the study aimed to contribute information to aid intervention program development by determining predictors of drink and drug driving behavior. The study examines both drug and alcohol use and the influence of attitudes toward driving while under the influence of alcohol and/or drugs. Results indicated that significant proportions of university students are engaging in the risk-taking behavior of driving under the influence of drugs (25%), alcohol (14%), or both (8%). The results suggest that attitudes toward driving under the influence of alcohol and/or drugs are a strong predictor of engaging in the behavior. Peer influence emerged as a major aspect of attitudes. The results of this study provide an important beginning to understanding drink and drug driving and the influence of attitudes in the university student body. PMID- 15823875 TI - Tire blow-outs and motorway accidents. AB - During the period from 1996 to 2002, 60,397 vehicles were involved in crashes with property damage and/or injury on a French motorway network of 2000 km. It was observed that 6.7% of these accidents involved tire blow-outs. In 87% of cases, only one vehicle was involved in the accident. Tire blow-outs occurred in 6.5% of cars that represented more than 80% of the vehicles involved in crashes. The occurrence of this phenomenon is very high for vans (22%), though it concerns trucks less (2.5%). The proportion of tire blow-outs decreased from 1997, when it was 8.0%, to 5.9% in 2002. However, two main facts require examination: (1) On inter-urban motorways, crashes involving blow-outs of rear tires occur four times more frequently than for blow-outs of front tires. (2) The frequency of tire blow outs is especially high for vans, and almost always involves rear tires. This higher frequency for rear tires is the result of two phenomena, which are indistinguishable given the data available: firstly, a four-wheel vehicle is more difficult to control if a blow-out occurs on a rear tire (confirmed experimentally); secondly, rear tires may be in poor condition more often than front tires, and so more prone to blow-outs. Consequently, users are strongly recommended to install the best tires on rear wheels. In practice, if only the front tires are replaced, which often occurs because they tend to be worn out more quickly than the rear ones (especially for front wheel drive vehicles), it is necessary to move the rear tires to the front and fit the new ones on the rear wheels. Very interesting technological developments are in progress that should reduce the number of tire blow-outs. However, considering the time necessary to renew the number of cars on the roads, this very simple and inexpensive recommendation should apply to all cars and especially to vans. PMID- 15823876 TI - Characteristics of vehicle-animal crashes in which vehicle occupants are killed. AB - During the past 10 years almost 1,500 people have been killed in motor vehicle collisions with animals. Police reports on 147 fatal vehicle-animal crashes during 2000-2002 were obtained from nine states. The goal was to determine common crash types, types of animals involved, and steps that could be taken to reduce the crashes and injuries. Seventy-seven percent of the struck animals were deer, but six other types of animals were involved including small ones such as dogs. Eighty percent of the crashes were single-vehicle events. In most of these cases a motorcycle struck an animal and the rider came off the vehicle, or a passenger vehicle struck an animal and then ran off the road; in a few cases the animal went through the windshield. Multiple-vehicle crashes included vehicles striking deer that went through the windshields of oncoming vehicles, vehicles striking animals and then colliding with other vehicles, and vehicles striking animals that subsequently were struck by other vehicles. Crashes occurred primarily in rural areas, on roads with 55 mph or higher speed limits, during evening or nighttime hours, and in darkness. Greater application of deer-vehicle collision countermeasures known to be effective is needed, but it is noteworthy that a majority of fatalities occurred from subsequent collisions with other vehicles or objects, not from animal contacts. Sixty-five percent of motorcyclists killed were not wearing helmets, and 60% of vehicle occupants killed were unbelted; many of these fatalities would not have occurred with proper protection. PMID- 15823877 TI - Motorcycle helmets and spinal cord injury: helmet usage and type. AB - The objective of this study was to assess the role of helmets and helmet type in relation to injury to the cervical spinal cord. It was based on a consecutive series of 110 motorcyclists with neurological damage to the spinal cord admitted alive (referred to as acute survivors) to a specialist spinal cord injuries unit at an Australian hospital. Cases were those with injury to the cervical spinal cord and controls were those with injury to the cord of other segments of the spine. The study showed that there was no significant difference in the odds of cervical spinal cord injury among unhelmeted and helmeted motorcyclist acute survivors. In addition, it confirmed the findings of a recently published Australian fatality study demonstrating no difference in the odds of cervical spinal cord injury among full-face and open-face helmet wearers. These results contrasted with the findings of earlier studies. In consideration of the limitations of existing research on the role of helmets in spinal cord injury, further study is required based on a larger series or a series having a higher proportion of non-wearers and open-face helmet wearers, including both survivors and those killed, and including assessment of cord and non-cord spinal injuries separately, helmet type, head impact, and helmet retention. PMID- 15823878 TI - The effect of muscle activation on neck response. AB - Prevention of neck injuries due to complex loading, such as occurs in traffic accidents, requires knowledge of neck injury mechanisms and tolerances. The influence of muscle activation on outcome of the injuries is not clearly understood. Numerical simulations of neck injury accidents can contribute to increase the understanding of injury tolerances. The finite element (FE) method is suitable because it gives data on stress and strain of individual tissues that can be used to predict injuries based on tissue level criteria. The aim of this study was to improve and validate an anatomically detailed FE model of the human cervical spine by implement neck musculature with passive and active material properties. Further, the effect of activation time and force on the stresses and strains in the cervical tissues were studied for dynamic loading due to frontal and lateral impacts. The FE model used includes the seven cervical vertebrae, the spinal ligaments, the facet joints with cartilage, the intervertebral disc, the skull base connected to a rigid head, and a spring element representation of the neck musculature. The passive muscle properties were defined with bilinear force deformation curves and the active properties were defined using a material model based on the Hill equation. The FE model's responses were compared to volunteer experiments for frontal and lateral impacts of 15 and 7 g. Then, the active muscle properties where varied to study their effect on the motion of the skull, the stress level of the cortical and trabecular bone, and the strain of the ligaments. The FE model had a good correlation to the experimental motion corridors when the muscles activation was implemented. For the frontal impact a suitable peak muscle force was 40 N/cm2 whereas 20 N/cm2 was appropriate for the side impact. The stress levels in the cortical and trabecular bone were influenced by the point forces introduced by the muscle spring elements; therefore a more detailed model of muscle insertion would be preferable. The deformation of each spinal ligament was normalized with an appropriate failure deformation to predict soft tissue injury. For the frontal impact, the muscle activation turned out to mainly protect the upper cervical spine ligaments, while the musculature shielded all the ligaments disregarding spinal level for lateral impacts. It is concluded that the neck musculature does not have the same protective properties during different impacts loadings. PMID- 15823879 TI - Effects of initial seated position in low speed rear-end impacts: a comparison with the TNO rear impact dummy (TRID) model. AB - Injury-producing mechanisms associated with rear-end impact collision has remained a mystery not withstanding numerous investigations devoted to its scrutiny. Several criteria have been proposed to predict the injury-causing mechanism, but none have been universally accepted. The challenge lies in determining a set of testing procedures representative of real-world collisions, wherein the results obtained are not only the same as human testing, but remain consistent with various subjects and impact conditions. It is hypothesized that one of the most important considerations in the testing methodology is the effect of initial seated position (ISP) on occupant kinematics during a rear impact collision. This study involves two parts that evaluates the effects of ISP during rear-end impact. In the first part, head acceleration results of computer simulation using Hybrid III TNO rear impact dummy (TRID) are compared to physical impact testing (PIT) of humans. The second part focuses on the computer simulation using TRID to obtain different neck parameters such as NIC (Neck Injury Criterion), NIJ (Neck Injury Predictor), neck forces and moments to predict the level of neck injury such as whiplash associated disorder (WAD) during low speed rear-end impact. In PIT, a total of 17 rear-impact tests were conducted with a nominal 8-km/hour change in velocity to 5 subjects in four different seated positions comprising of a normal position (NP) and three out of positions (OOP). The first position was a NP, defined as torso against the seat back, looking straight ahead, hands on the steering wheel, and feet on the floor. The second position was a head flex position (HFP), defined as the normal position with head flexed forward approximately 20 degrees. The third position was a torso lean position (TLP), defined as the normal position with torso leaned forward approximately 10 degrees away from the seat back. Lastly, a torso lean head flex position (TLHFP), defined as the normal position with the head flexed forward approximately 20 degrees and torso leaned forward approximately 10 degrees. The head acceleration plots from PIT reveal that for the third and fourth positions (TLP and TLHFP) when the subject torso leaned forward, the peak head acceleration for the subject decreased and there was also a delay in reaching the peak. The Hybrid III-TRID anthropomorphic test dummy (ATD) was used in the same four different seated positions using computer simulation software MAthematical DYnamic MOdel (MADYMO 6.0) and the head acceleration results were compared to PIT. The comparison demonstrates that the Hybrid III-TRID ATD with MADYMO can be a reliable testing procedure during low-speed, rear-end impact for the four ISPs considered since the head acceleration plots deviated within the range of PIT head acceleration plots for different human subjects. This ensures that the second part of the study with neck injury using computer simulation results is a reliable testing procedure. It can be observed that MADYMO results have a greater error when compared to PIT when more than one OOP condition is employed as in TLHFP. All these observations would help in providing a tool to better understand the injury mechanisms and provide an accurate testing procedure for rear-end impact. PMID- 15823880 TI - Risk curve boundaries. AB - Insights into the science and the dynamics associated with the stimulus-injury relationship of the human system are often gained by focusing on the results obtained from human surrogate testing. These results are commonly mapped into an injury risk paradigm for the purpose of characterizing the stimulus injury response: the injury risk curve. However, the quality and quantity of the data along with the experimental design are critical factors when considering the value of the estimated risk curve. Presented in this article is an analysis of injury risk curves in terms of their usability and appropriateness with respect to the sample size, stimulus distribution over the critical range, censoring, shape of the underlying risk function, and the inclusion of "actual" (uncensored) along with censored data. The results from this analysis indicate that for "large" biomechanical injury data sets there is no advantage to using actual data; censored data will yield the same injury risk curve as actual data. However, for "small" biomechanical injury data sets the inclusion of actual data will significantly improve the quality of the resulting risk curve. In addition, the results show that the amount of injury data needed to generate a risk curve with a given confidence bound is not only dependent on the relative contribution of the censored data and actual data but also on the shape of the risk function along with the stimulus distribution over the critical range. Confidence intervals are presented for the thoracic injury risk and the head injury risk to show the influence of data distribution on the goodness of the risk function estimation. PMID- 15823883 TI - Pervasive alienation: on seeing the invisible, meeting the inaccessible and engaging 'lost to contact' clients with major mental illness. AB - Increasing integration of services for adults with severe mental health problems has highlighted the challenges to services posed by clients who are difficult to engage. There is a dearth of studies on direct work with individuals who are reclusive and difficult to contact, in contrast with the bulk of literature where the patient and network are present and available. This paper describes the work of a busy CMHT with such clients; barriers to collaborative work are explored and some of the team's strategies to overcome these barriers are delineated. Long, slow and persistent contact is observed to be a core feature of the work. The paper goes on to examine the fit between the realities of ongoing work with such clients and the available therapeutic and service discourses. PMID- 15823884 TI - Interprofessional teamworking for child and family referral in a Sure Start local programme. AB - Interprofessional and inter-agency working are important features of UK government initiatives, such as Sure Start local programmes for children under 4 years old and their families. Part of the vision for Sure Start was that providers of services and support would work together in new ways that cut across old professional and agency boundaries and focus more successfully on family and community needs. This paper describes the development and functioning of a Referral and Allocation Project in one trailblazer Sure Start local programme. The Programme employs staff with backgrounds that include health, social work, education and clinical psychology who work in a shared location. The Referral and Allocation Project sought to develop, through regular meetings, a whole-team inter-agency focus on discussing the needs of families who had been referred, or had referred themselves, to Sure Start and on suggesting ways in which support or advice could be offered and accessed in order to meet their needs. The paper examines issues that arose during the development of the Project, in particular psychodynamic dimensions such as emotional impact and professional anxiety. These may have implications for the format and culture of interprofessional team meetings and for training and professional development. PMID- 15823885 TI - Team types, perceived efficiency and team climate in Swedish cross-professional teamwork. AB - This paper aims to identify the dominant types of team organization in cross professional Swedish human service organizations and the relationship between team type and perceived efficiency as well as team climate as an aspect of work satisfaction. A questionnaire was responded to by 337 individual professionals from 59 teams, mainly from psychiatric care (50.7%) but also from social, neuropaediatric and vocational (re)habilitation, school health care and the occupational health service. The interprofessional model of team organization was the most frequent (62%), followed by the transprofessional (33%), and the multiprofessional team, (5%). A moderate positive correlation was found between team type and perceived efficiency as well as team climate. The greater the interdependence and the closer the co-operation, the higher the efficiency and the better the climate. No differences were found between professions or organizational domiciles with respect to team type. This paper suggests (1) a more consistent vocabulary with 'cross-professional' as the generic term covering different team types and (2) that a contingency approach to teamwork is tested in future research. PMID- 15823886 TI - Interdisciplinary collaboration between primary care, social insurance and social services in the rehabilitation of people with musculoskeletal disorder: effects on self-rated health and physical performance. AB - Previous research shows there can be good results from co-financing between welfare sectors on the perceived quality of interprofessional collaboration. However, little is known about the impact on patient outcome of such schemes. This study aimed to assess whether co-financed teams with personnel from primary care, social insurance and social services have any effect on patients' health status. A comparative study of patients attending health care centres with and without a co-financed collaboration model was carried out. Although research has shown positive results from co-financed collaboration on staff and organization, we could not find that this new interdisciplinary team structure gave a better patient health outcome than conventional care. PMID- 15823887 TI - Evaluation and development of integrated teams: the use of Significant Event Analysis. AB - Significant Event Analysis (SEA) is a practical, problem-solving technique that can help teams learn from their experiences and generate practical solutions to complex issues. It has mainly been conducted in Primary Health Care Teams (PHCTs) with the use of an external facilitator. Although SEA is envisioned to address issues of integration and complement routine audit, there is limited evidence that this has been achieved. This paper reports on a 1 day workshop during which team members were trained to use SEA to address issues of service integration. This paper presents an evaluation of the workshop in order to determine if SEA is a feasible tool to support and encourage team self-evaluation without external facilitation. The main barriers to SEA are discussed as well as the need for longitudinal research to assess the impact of SEA on integration of the team and wider service. PMID- 15823888 TI - The place of 360 degree appraisal within a team approach to professional development. AB - The role of appraisal within professional development has assumed a higher profile in response to national policies including those relating to clinical governance and workforce development. Under the guidance of a service development manager trained in human resource management, a new interprofessional Primary Care Practice Team near Cambridge defined and implemented a new appraisal system drawing on the principles of 360 degree appraisal. The appraisal system was designed to support the service function and the team culture, and took into account good management practice and the requirements of partner organizations and professional affiliations. This paper describes how the system was introduced, tailored to the particular needs of the interprofessional practice team, and evaluated. Early experience suggests that the approach enhances understanding of roles and responsibilities and is supportive to interprofessional team development. It is well received by team members involved and by some of the external partner organizations. However, it is demanding on resources and concerns were raised about the completeness of this approach to appraisal. PMID- 15823889 TI - Quality Practice Award and teamworking: the perceptions of Primary Health Care Team members in Scotland. AB - Evidence exists that those teams which complete an agreed task experience an improvement in team functioning. The aim of this study was to investigate whether completion of the Royal College of General Practitioners Quality Practice Award (QPA) improved the team members' perception of teamworking within their Primary Health Care Teams (PHCTs). A postal questionnaire was sent to all 158 PHCT members in 6 Scottish practices that achieved QPA in 2001 followed by selected telephone interviews. General practitioners (GPs), nurses and administrative staff showed a statistically significant improvement in their perception of teamworking as a result of completing QPA, but nurses reported this to a lesser degree. Protocols and audit work, followed by improvements in communication were perceived to have had the most beneficial effect on teamwork. After completing QPA, 79% of GPs, 64% of nurses and 84% of administrative staff perceived an improvement in the standard of care provided. Qualitative data confirmed that team functioning improved by completing QPA, but that a certain level of team functioning was necessary before a practice begins the QPA process. Teams also hoped to maintain the high standards inherent in QPA in the future but had concerns regarding workload. PMID- 15823890 TI - Multidisciplinary perspectives on core networking skills. A study of skills: and associated training needs, for professionals working in managed clinical networks. AB - Possessing a wide mix of non-clinical competences is important for professionals involved in managed clinical networks (MCNs). Skills that stand out are related to interpersonal issues, problem solving, decision-making, and managing change. Interprofessional and interorganizational collaboration is important in health care generally and is not confined to MCNs. Skills are likely to have relevance in wider contexts. Training needs identified for professionals in MCNs relate to skills associated with working in challenging situations, including: 'managing change,' 'conflict resolution,' and 'negotiation.' Limited generalizations about profession-specific skills and training needs can be made. However, it is more appropriate to identify skills needed for the specific role(s) an individual is asked to perform, and to investigate if there are performance gaps between skills and competencies. PMID- 15823891 TI - Interdisciplinary working between community pharmacists and community nurses: the views of community pharmacists. AB - Changing roles within primary care have been heralded by the implementation of nurse prescribing and the need to maximize skills within the existing workforce. This paper describes the views of community pharmacists regarding nurse prescribing and interdisciplinary working in primary care. Limited contact was reported between community pharmacists and prescribing nurses which contrasted with the reported frequency of contact with other health professionals. The community pharmacists reported positive views regarding teamworking and nurse prescribing which it is suggested provides a readiness to develop interdisciplinary working which hitherto had foundered on lack of contact and in consequence insufficient understanding of roles and a shared view of primary care. PMID- 15823892 TI - The effect of community-based, interdisciplinary training on the careers of physicians. PMID- 15823896 TI - The bicycle makes the eyes smile: exercise, aging, and psychophysical well-being in older Italian cyclists. AB - Despite a cultural and biomedical consensus about the health benefits of physical activity, most adults do not exercise regularly and their activity tends to decline further in older age. This study reports on a group of older-age Italian bicyclists who maintain extraordinary activity levels. It explores the physical and mental health effects of intensive exercise in older people, the meanings and motivations associated with it, and the cultural and social features that support and promote bicycling in Italy. The findings suggest some public health implications for the prevention of chronic diseases caused by inactivity and excessive body weight. PMID- 15823897 TI - The role of ethics in commercial genetic research: notes on the notion of commodification. AB - The emergence of exchange systems for new bodily entities such as organs, cell lines, and tissue samples has generated increasing ethical concern. Concurrently, the role of ethics is becoming contested. Some social scientists have sought to reveal ethics policies as veils for commercial exploitation, masking the crude commodification of the human body. Other social scientists and ethicists have attempted to carve out a role for ethics as a defense against such, presumably violent, market forces. This article seeks an alternative rout. In analyzing the commercialization of a particular Swedish biobank, it suggests that we are in fact witnessing a process of decommodification and that ethics policies do indeed play a central role in this. However, concomitantly, ethics can be seen to veil the commodification not of bodily entities but, rather, of research results. When approaching these processes it is suggested that we analyze the interrelatedness of moral reasoning and forms of exchange. PMID- 15823898 TI - Saving Tibet? An inquiry into modernity, lies, truths, and beliefs. AB - Social theorists have explored the ways in which quantification serves as an instrument of governance in the modern state, whether tied to concerns of population size and quality or to problems of social behavior. Biopolitics are as visible in the modern socialist states as they are in free-market democratic states, and they are perhaps nowhere more visible today than in the new global standards of "evidence-based medicine," wherein it is assumed that only quantifiable evidence can serve to establish policy, procedure, and outcome. When it comes to creating ways to "civilize" and organize their target citizenry through health development, Socialist China as relied on such technologies as much as have health development funding agencies from donor countries. In this article, I look at quantitative methods in relation to assumptions that morality can be severed from truth and that numbers are potentially morally neutral. This idea is tied not only to forms of modern subjectivity but also to the distinct ways in which certain linguistic and theoretical practices relate to provisional notions of "lying," "truth-telling," and ways of "believing." An exploration of the effects of attempts to quantify maternal behavior, morbidity, and morality in rural Tibet highlights the problem of morality within an environment in which numbers are never free-standing but, rather, are always presumed to carry moral messages, and in which domains that cannot be quantified serve as a primary basis for knowing truth. Through an exploration of rural Tibetan encounters with health development programs for safe motherhood, I provide a critique of quantification and return to questions about "belief" as a rubric that interrupts modern dichotomies of lies versus truths. PMID- 15823900 TI - Phototherapy with narrowband vs broadband UVB. AB - Phototherapy with ultraviolet (UV) radiation of wavelengths between 280 and 320 nm (UVB) is a safe and effective treatment for a variety of diseases. In addition to standard broadband UVB (bUVB), narrowband phototherapy with fluorescent bulbs emitting near monochromatic UV around 311 nm (nUVB) has become an important treatment for diseases such as psoriasis, atopic dermatitis and vitiligo. In addition to these indications, the number of diseases for which nUVB phototherapy is reported to be effective is continuously growing. The differential effects of nUVB phototherapy in comparison to other UV wavelengths as well as established and new indications for this treatment modality are reviewed. PMID- 15823901 TI - Cytokine gene polymorphisms in patch-stage mycosis fungoides. AB - Cytokine production is under genetic control and certain allelic variants of cytokine genes are associated with lower or higher cytokine production in vitro and in vivo. The general concept is that a shift from a Th1 to a Th2 cytokine profile accompanies disease progression from patch-stage mycosis fungoides to tumour stage, although the results of the studies carried out have not been entirely conclusive. We aimed to investigate whether certain cytokine polymorphisms might represent a risk factor for developing patch-stage mycosis fungoides. Genotyping for IFN-gamma (Th1 cytokine), IL-6, IL-10 (Th2 cytokines), TNF-alpha and TGF-beta 1 was undertaken for 33 patients with patch-stage mycosis fungoides and the results were compared with those in a control group. Genotype distribution showed no significant differences between the patients and the controls for any of the five cytokines studied. Our study suggests that patch stage mycosis fungoides is not determined by a specific genotype polymorphism. However, further studies on larger numbers of cases are needed before definite conclusions can be drawn. PMID- 15823902 TI - Selective persistence of dermal CD8+ T cells in lesional plaque psoriasis after clobetasol-17 propionate treatment. AB - In psoriasis, T-cell infiltration and epidermal hyperproliferation are key phenomena which are closely related. Our aim was to investigate the dynamics among T-cell subsets in relation to epidermal proliferation and clinical severity in psoriasis during treatment with an ultra-potent corticosteroid. Seven psoriasis patients were treated twice daily for 14 days with clobetasol-17 propionate ointment. Punch biopsies were taken at day 0, 3, 7 and 14. Epidermal proliferation marker Ki-67 and CD4+, CD8+, CD45RO+, CD2+ T cells were quantified by immunohistochemical techniques and image analysis. The clinical score declined significantly (60%; p<0.01) and a 47% reduction of Ki-67+ nuclei was observed after only 3 days (p<0.01). In the epidermis all investigated T-cell subsets were significantly reduced at day 14 (p<0.05). In the dermis, treatment resulted in a significant decrease of CD4+, CD45RO+ and CD2+ T cells, but dermal CD8+ T cells persisted. In psoriasis, reduction of clinical severity and epidermal proliferation during the early phase of topical corticosteroid therapy cannot primarily be the result of decreased T-cell subsets. Furthermore, selective persistence of dermal CD8+ T cells was observed, which might be associated with disease relapse. PMID- 15823903 TI - The vulvar skin microenvironment: impact of tight-fitting underwear on microclimate, pH and microflora. AB - The aim of the present study was to investigate if tight-fitting underwear (string panties) equipped with string panty liners affected the vulvar skin microenvironment differently to regular panties with standard panty liners. Thirty-two healthy women participated in a crossover study where temperature, humidity, surface pH and aerobic microflora were measured on vulvar skin. Vulvar skin temperature was 35.2 +/- 0.19 (mean +/- SEM) and 35.3 +/- 0.17 degrees C, respectively, for the two underwear systems. Mean humidity and mean skin surface pH at vulvar skin did not differ between the two systems. Barely noticeable differences were found for the aerobic microflora both at labium majus and at perineum. The mean total number of microorganisms in the two different panty liners was the same, 6.0 +/- 0.15 and 6.0 +/- 0.16, respectively (log CFU per panty liner). The differences in panty and panty liner design studied seem to have negligible impact on the vulvar skin microclimate, skin surface pH and aerobic microflora. No support was found for the assumption that a string panty system would result in higher contamination of vulvar skin by anorectal microflora. PMID- 15823904 TI - Atopy patch test reactions to house dust mites in patients with scabies. AB - It is well known that the house dust and the scabies mites are related phylogenetically. We therefore performed atopy patch tests with house dust mite antigens (Dermatophagoides pteronyssinus (Dp) and/or Dermatophagoides farinae (Df)) in scabies patients without atopy and healthy controls. We studied 25 men with active scabies and 25 healthy controls. Skin prick tests with standardized house dust mite extract were performed for all patients and controls. An intradermal test procedure was carried out in skin prick test-negative patients, and for controls showing positive atopy patch test to Dp and/or Df. While atopy patch tests were performed directly in all healthy controls, patients with scabies were first treated and on the next day, atopy patch tests were performed. Twenty-two of 25 patients with scabies (88%) had skin prick test and/or intradermal test positivity against house dust mites, whereas 17/25 patients (68%) had atopy patch test positivity against house dust mites (Dp and/or Df). There was no statistically significant difference between skin prick test and/or intradermal test positivity and atopy patch test positivity in a regression analysis (p=0.222). The only statistically significant correlation was between atopy patch test positivity and the extent of scabies involvement (p<0.05). Only few of the healthy controls had positive tests. In this study, we have shown that a positive atopy patch test to house dust mite antigens is not specific for patients with atopic dermatitis, but also occurs in scabies patients without a history of atopic dermatitis. PMID- 15823905 TI - Rearing larvae of Lucilia sericata for chronic ulcer treatment--an improved method. AB - Larval therapy is a biological 'dressing' for chronic ulcers which is used for debridement, to initiate healing and to reduce bacteria and unpleasant odour. One condition for being able to employ larval therapy is access to disinfected larvae of good quality. We therefore started rearing Lucilia sericata larvae in 1998. Rearing necrophagous larvae in a hospital setting makes it essential to control odour. The improvement of the method includes the use of a controlled, humid, warm environment with artificial light and inexpensive disposable material, as well as the use of ready-made, constantly available and aseptically produced nutrition for feeding larvae and flies. With equipment such as a refrigerator and freezer, larval food is kept fresh and odour-free and chloramine solution is used to disinfect the eggs and thus the larvae. Adhering to a proposed weekly working schedule makes the rearing procedure effective and reliable. PMID- 15823906 TI - Severe impairment of quality of life in Hailey-Hailey disease. AB - Hailey-Hailey disease (a skin fragility disorder) runs a chronic course and may cause important disability. However, little has been formally investigated concerning the quality of life (QoL) of patients affected by this disorder. We studied the impact of Hailey-Hailey disease on the QoL of 22 consecutive patients seen at our dermatological clinic. Patients were examined by a dermatologist, and they were asked to complete two self-administered questionnaires: the Skindex-29, in order to determine their QoL, and the 12-item General Health Questionnaire, to evaluate psychological distress. Completed questionnaires were returned by 20 patients. Even in patients with few body sites involved, median overall QoL scores were much higher than those observed in other skin conditions in all three domains investigated by the Skindex-29 (i.e. symptoms, emotions and social functioning). This was true also for the levels of psychological distress. QoL impairment was substantial irrespective of the number of body sites involved. Our findings document a great impact of Hailey-Hailey disease on patients' QoL. Therefore, a more aggressive therapeutic approach may be warranted in all patients, including those with few lesions. PMID- 15823907 TI - A retrospective epidemiological study on the association of bullous pemphigoid and neurological diseases. AB - Bullous pemphigoid is a rare chronic recurrent dermatosis that is often reported in association with various neurological diseases. No investigation involving a large number of patients has ever been carried out to demonstrate such an association. This study was accomplished by analysing the discharge diagnosis of all hospitalized patients, both day-patients and inpatients, during a 5-year period (1995-2000) covering a total population group of 934,023 living in a region of Italy that has approximately 1,200,000 inhabitants. The results support the hypothesis of an association between bullous pemphigoid, multiple sclerosis and Parkinson's disease on a highly significant statistical basis. The aetiopathogenic mechanisms and the causes that induce the loss of immunological tolerance are not yet understood. PMID- 15823908 TI - The occurrence of atopic dermatitis in Greenland. AB - Until recently there was no information available on the prevalence of atopic dermatitis in Greenland. Our objective was to determine the prevalence of atopic dermatitis in younger schoolchildren in Greenland. In the autumn of 2000 we used our previously elaborated questionnaire in a cross-sectional study of 954 schoolchildren aged 7-8 years, who lived in five Greenlandic towns. The findings were compared to data on atopic dermatitis from Denmark. The response rate was 65% (622). The lifetime prevalence of atopic dermatitis was calculated to be 14.0% (95% confidence interval 11.3-16.7) using our standard score criteria with an absolute lower limit estimate of 4.5%. Taking the response rate of 65% into consideration this study indicates that in 2000 the lifetime prevalence of atopic dermatitis among younger schoolchildren in Greenland was in the range of 10-15%. PMID- 15823909 TI - Optimized UVB treatment of atopic dermatitis using skin reflectance measurements. A controlled, left-right comparison trial. AB - In a randomized, open, left-right comparison study, 20 patients with atopic dermatitis were treated with UVB. One side of the body received UVB in a conventional regimen with fixed dosage increments, the other side was given UVB dosages according to skin reflectance measurements of skin pigmentation and erythema. Clinical outcome was assessed by SCORAD. The initial, final and cumulative UVB dosages, time to 50% reduction in SCORAD, and side effects were compared. The initial UVB dosage was somewhat higher in the skin reflectance guided treatment than in the conventional UVB regimen, although not significantly. There was no difference in the reduction of SCORAD comparing the two treatment options; however, the final UVB dosage and the cumulative UVB dosages were significantly lower in the optimized regimen. This new technique offers the same therapeutic advantage and security as a dose regimen guided by minimal erythema dose testing. However, measurement of skin pigmentation by skin reflectance is a rapid method, which can easily be operated by nurses. PMID- 15823910 TI - Atopy patch test--reproducibility and elicitation of itch in different application sites. AB - We evaluated the reproducibility of atopy patch test reactions and the quality and quantity of itch in 16 patients with atopic eczema and a history of a positive atopy patch test reaction, comparing three different application sites. The allergen was re-applied simultaneously on both forearms and the back. Intensity and quality of pruritus were evaluated using a visual analogue scale and the Eppendorf itch questionnaire, respectively. The atopy patch test reaction was highly reproducible, occurring in 15/16 (94%) patients. Pruritus was reported by 14/16 (88%) patients. There was no significant difference in either the intensity or quality of itch between the two forearms and the back (p>0.05). The mean peak visual analogue scale itch score was comparable across all three test sites (range 28.3-31.9). Regarding quantification of test reactions, a positive reaction was more frequent on the back (94% versus 69% on the arms) and the peak atopy patch test score was higher on the back compared with the arms (right forearm, p=0.0018 and left forearm, p=0.0683). Allergens should preferably be applied on the back for the atopy patch test. However, the atopy patch test can induce atopic itch irrespective of the application site. PMID- 15823911 TI - KID Syndrome: report of a Scandinavian patient with connexin-26 gene mutation. AB - Keratitis-ichthyosis-deafness syndrome is a rare genodermatosis, which has recently been connected with mutations in the connexin-26 gene, GJB2. We present a 15-year-old boy with erythroderma, hyperkeratotic plaques and deafness. Sequencing analysis showed a heterozygous missense mutation D50N (148G>A) in GJB2. The boy has not yet manifested characteristic eye lesions but his case shows that tardy development of eye signs should not preclude a clinical diagnosis of keratitis-ichthyosis-deafness syndrome. Besides the typical clinical features, the patient's height was above the 98th percentile and he displayed a delayed bone age in his hands. Additionally, he suffered from migrainoid headaches and the results of a magnetic resonance scan of the cerebrum showed he had a large cisterna magna which probably occurred independently from the syndrome. This patient is the first Danish patient in whom the keratitis ichthyosis-deafness syndrome has been verified by mutation analysis. PMID- 15823912 TI - Cutaneous Scedosporium apiospermum infection in an immunocompromised patient and a review of the literature. AB - Scedosporium apiospermum (also known as Pseudallescheria boydii) is a ubiquitous filamentous fungus. This fungus is known as a cause of mycetoma, which may occur in a normally immune host following trauma. However, in an immunocompromised host, S. apiospermum may cause a life-threatening infection. We describe a case of S. apiospermum infection of the right hand in a patient who was receiving long term immunosuppressants for adult Still's disease. We also review the cases of S. apiospermum infection with cutaneous manifestations reported between 1998 and 2003. PMID- 15823913 TI - Papillary endothelial hyperplasia and dilated lymphatic vessels in bullous pilomatricoma. AB - This is a report of papillary endothelial hyperplasia in a 9-year-old girl with a pilomatricoma showing bullous appearance. Histologically, papillary proliferation of endothelial cells was found within dilated lymphatic endothelium-lined vascular channels overlying a pilomatricoma. The endothelial cells covering the papillae were of a lymphatic endothelial cell nature proved by immunohistochemistry and electron microscopy. Abundant fibrous long-spacing collagen was observed in the connective tissue and fibroblasts within papillae. PMID- 15823914 TI - The prevalence of hepatitis C in patients with porphyria cutanea tarda in Stockholm, Sweden. AB - In many countries hepatitis C virus infection has been considered a major factor triggering overt porphyria cutanea tarda. The prevalence of hepatitis C virus infection was retrospectively studied in 87 patients who during a period of 11 years were diagnosed with porphyria cutanea tarda in Stockholm. Among patients with the sporadic form of porphyria cutanea tarda, the prevalence of hepatitis C virus infection was 36.4%. As hepatitis C virus infection may today be successfully treated and as the infection may be clinically silent and thus unknown to the patient, it is important to screen all patients with porphyria cutanea tarda for hepatitis C virus infection. PMID- 15823915 TI - Infectious urticaria with purpura: a mild subtype of urticarial vasculitis? AB - Urticaria is characterized by transient wheals. We report here five cases with long-lasting urticarial lesions persisting for more than 24 hours. Each lesion left purpura after fading. There was no systemic involvement. C-reactive protein and serum levels of complement were elevated or normal. Histologically, marked infiltration by eosinophils and neutrophils with karyorrhexis in the perivascular and intercollagenous spaces was observed, but there was no evidence of vasculitis (venulitis). Skin symptoms were resistant to systemic corticosteroids. In contrast, treatment of underlying bacterial infections resulted in marked improvement of skin lesions. E-selectin, VCAM-1 and ICAM-1 were expressed on endothelial cells. Marked deposition of C3a, C5a, neutrophil elastase and major basic protein in the dermis was observed. These urticarial lesions provoked by bacterial infections seem to lie on the continuum between urticaria and urticarial vasculitis. PMID- 15823916 TI - Dissection of antigen-specific humoral and cellular immune responses for the development of experimental immune-mediated blepharoconjunctivitis in C57BL/6 mice. AB - PURPOSE: Allergic conjunctivitis is characterized by allergen-specific IgE in the serum and infiltration of eosinophils into the conjunctiva. However, it remains unclear whether early-phase reaction (EPR) mediated by Ag-specific IgE links to late-phase reaction (LPR) in the conjunctiva. We aimed to investigate whether LPR is mediated by either cellular or humoral immune responses. METHODS: Experimental immune-mediated blepharoconjunctivitis (EC) was induced in C57BL/6 mice by either active immunization or passive immunization by transfer of ragweed (RW)-primed lymphocytes and RW-specific IgE, followed by RW challenge onto the conjunctiva. Transferring RW-primed lymphocytes were prepared from RW-primed splenocytes which were stimulated in vitro with RW for 3 days. Fifteen minutes after RW challenge, clinical findings were evaluated and 24 hr after challenge, the conjunctivas and sera were harvested for histologic analysis and measurement of IgE, respectively. RESULTS: EPR was most prominent when EC was induced by transfer of RW-specific IgE. EPR was hardly detectable if EC was induced by transfer of RW-primed lymphocytes. Mild EPR was noted when EC was induced by active immunization. LPR, evaluated by infiltration of eosinophils into the conjunctiva, was most severe when EC was induced by transfer of RW-primed lymphocytes. Minimal, but definite LPR was induced when EC was induced by transfer of RW-specific IgE. Intermediate severity of LPR was induced when EC was induced by active immunization. CONCLUSIONS: LPR in the conjunctiva is dominantly mediated by cellular immune responses, whereas EPR in the conjunctiva is putatively mediated by humoral immune responses. Importantly, LPR in the conjunctiva is inducible by Ag-specific IgE alone, although minute. PMID- 15823917 TI - A single local injection of recombinant VEGF receptor 2 but not of Tie2 inhibits retinal neovascularization in the mouse. AB - PURPOSE: The purpose of this study was to develop pharmacological therapeutic alternatives for ischemia-induced proliferative retinopathy. METHODS: C57BL/6J mice were placed in 76% oxygen on postnatal day 7 (P7) for 5 days. On P12 recombinant, chimeric vascular endothelial growth factor (sVEGF-R2) or sTie2 was injected intravitreally in one eye. The fellow eye received a control injection. On P17, retinal wholemounts were prepared after perfusion with fluorescein dextran to quantify the retinopathy. RESULTS: A single intravitreal injection of sVEGF-R2 reduced pathologic vascular changes significantly (p = 0.02). No significant effect was observed after intravitreal application of sTie2 (p = 0.07), although Ang-2 was upregulated in control animals without treatment as neovascularization developed and Ang-1 was constantly transcribed (ratio PCR). CONCLUSIONS: sVEGF-R2 interferes with VEGF signaling via VEGF-R2 receptor. Thus, local application of soluble receptors for angiogenic factors is a possible therapy for proliferative retinopathy. Receptors with a wide range of ligands might prove more useful for local application than those binding few or antagonistic ligands. PMID- 15823918 TI - Retinal vascular development and pathologic retinal angiogenesis are not impaired in matrix metalloproteinase-2 deficient mice. AB - PURPOSE: Earlier studies have suggested a role for metalloproteinase-2 (MMP-2) in retinal angiogenesis. To investigate this further, we have studied retinal vascular development and pathologic ischemia-induced retinal angiogenesis in MMP 2-deficient and wild-type mice. METHODS: Vascular development of the retina was studied in retinal flatmounts, whereas pathologic retinal angiogenesis was analyzed in retinal flatmounts and on histologic sections using a model of ischemia-induced retinopathy. The time course of MMP-2 mRNA expression was determined by in situ hybridization and real-time polymerase chain reaction (PCR). RESULTS: Formation of the retinal vascular plexus was not significantly different in MMP-2-deficient mice as compared to wild-type mice. In ischemia induced retinopathy, there was an increased formation of extraretinal neovascular tufts in the MMP-2-deficient mice (p < 0.05). MMP-2 mRNA expression did not correlate to either retinal vascular development or to ischemia-induced formation of extraretinal vascular tufts. CONCLUSIONS: The current data suggest that MMP-2 is not essential for either retinal vascular development or pathologic retinal neovascularization in the mouse. PMID- 15823919 TI - Molecular genetic and ocular findings in patients with holt-oram syndrome. AB - PURPOSE: The autosomal dominant Holt-Oram syndrome (HOS) is characterized by upper limb and cardiac septal defects. Mutations of the TBX5 gene have been identified as the underlying gene defect in HOS. Embryonic expression of TBX5 has been found in the human retina. This is the first report of ocular findings in two unrelated families with mutations in the TBX5 gene. METHODS: Six living persons affected with HOS and 10 unaffected family members were subjected to mutation analysis and complete ophthalmological examination, including electrophysiological examinations (EOG and flash ERG). RESULTS: A heterozygous single base-pain substitution in exon 5 (408C --> A) was detected in all affected patients. All examined affected patents were ophthalmological asymptomatic with normal EOG. A scotopic elongated b-wave latency was found in affected family members who were older than 35 years. The ERG was normal in the young patients. CONCLUSIONS: Haploinsufficiency of TBX5 alters the dorsal-ventral polarity in developing eye vesicles without amy detected functional loss in human. Slight ERG abnormalities later in life may be a result of changes induced by the inner ganglion cell layer in the inner nuclear layer. PMID- 15823920 TI - Hereditary high hypermetropia in the Faroe Islands. AB - PURPOSE: To characterize the phenotype of two families with high hypermetropia from the Faroe Islands. METHODS: Ophthalmologic evaluation including ultrasound oculometry and anthropometric measurements. RESULTS: Of the 40 examined family members, 15 individuals (8 males, 7 females; ages: 6-77 years; mean: 36.5 years) had small deep-set eyes with high hypermetropia (median: + 16.5 D; range: + 7.75 to + 22), short axial eye length (< 21 mm), and a thickened eye wall. The median corrected visual acuity was 0.4 (0.2-0.9). Ocular complications included angle closure glaucoma in six eyes, uveal effusion in three eyes, cataract in two eyes, and esotropia with amblyopia in three eyes. An emergency case of uveal effusion and retinal detachment after Yag iridotomy eventually responded to systemic corticosteroids and scleral resection surgery with a slow visual recovery. No associated ocular or systemic malformations were found in the series. In addition to the two examined families, six smaller Faroese families with high hypermetropia are briefly reported. CONCLUSIONS: The study highlights the signs and symptoms of a rare hereditary phenotype characterized by a short axial length mainly confined to the posterior segment of the eye, a shallow anterior chamber, and a thickened eye wall. The morphological characteristics predispose for sight threatening complications such as angle-closure glaucoma, chorioretinal pathology including uveal effusion, and amblyopia. Regular ophthalmic follow-up is therefore of obvious importance in families known to have small eyes/high hypermetropia. An endemic high prevalence in the Faroe Islands suggests the presence of a founder effect, and further genetic research would probably indicate pseudodominant rather than dominant transmission PMID- 15823921 TI - Penetrance and phenotype of the Thr377Met Myocilin mutation in a large Finnish family with juvenile- and adult-onset primary open-angle glaucoma. AB - PURPOSE: To study the role of myocilin (MYOC) as a susceptibility gene for juvenile- and adult-onset open-angle glaucoma (JOAG and POAG, respectively). METHODS: In a six-generation Finnish family with JOAG and POAG, we performed thorough ophthalmologic characterization (including assessment of the visual fields by Octopus perimetry, nerve-fiber layer thickness by photography, and disc size by Heidelberg tomography) of 51 individuals. The coding region of MYOC was screened for mutations by PCR amplification and direct sequencing. RESULTS: We detected a C > T transition at codon 377 resulting in a substitution of a threonine residue for methionine (Thr377Met) in the olfactomedin-like domain of myocilin, segregating in the family. Of the 20 individuals heterozygous for the mutation, nine (45%) were glaucomatous and two (10%) had ocular hypertension (OHT). The mean age at diagnosis of glaucoma in these individuals was 34.3 years (range: 14-66 years). Moreover, three of these individuals suffered retinal vein occlusion (RVO) in one eye, while one individual without the mutation had RVO. CONCLUSION: Our results further support the evidence that the Thr377Met mutation in MYOC may represent a susceptibility allele for glaucoma. These findings may facilitate genetic counseling, and early diagnosis and treatment of glaucoma. The possible interaction of factors contributing to RVO in conjunction with the Thr377Met mutation warrants further investigation. PMID- 15823922 TI - Novel mutations in MYO7A and USH2A in Usher syndrome. AB - PURPOSE: Usher syndrome is an autosomal recessive disease associating retinitis pigmentosa and neurosensory deafness. Three clinical types (USH1, USH2, USH3) and 11 mutated genes or loci have been described. Mutations in MYO7A and USH2A are responsible for about 40% and 60% of Usher syndromes type 1 and 2, respectively. These genes were screened in a series of patients suffering from Usher syndrome. METHODS: We performed SSCP screening of MYO7A in 12 unrelated patients suffering from Usher syndrome type 1 (USH1) and USH2A in 28 unrelated patients affected by Usher syndrome type 2 (USH2). RESULTS/CONCLUSIONS: Six mutations in MYO7A were found in five patients, including two novel mutations c.397C > G (His133Asp) and 1244-2A > G (Glu459Stop), accounting for 42% of our USH1 patients. Twelve mutations in USH2A were found in 11 patients, including four new mutations c.850delGA, c.1841-2A > G, c.3129insT, and c.3920C > G (Ser1307Stop), accounting for 39% of our USH2 patients PMID- 15823923 TI - Mitochondrial T9957C mutation in association with NAION and seizures but not MELAS. AB - PURPOSE: To assess the functional significance of the mitochondrial nt-9957 mutation in a man with non-arteritic ischemic optic neuropathy (NAION). This nt 9957 mutation has been previously reported in association with mitochondrial encephalopathy, lactic acidosis, and stroke-like events (MELAS). METHODS: The patient was examined clinically and with magnetic resonance imaging and spectroscopy. The entire coding region of the mitochondrial genome was sequenced, and mitochondrial function was assessed by flow cytometry after staining with fluorescent dihydroethidium. RESULTS: This 76-year-old man had optic nerve disease bilaterally and seizures, but no clinical or radiological evidence of MELAS. He had no mitochondrial DNA mutation other than the 9957. Functional testing revealed a severe defect in mitochondrial complex III activity. CONCLUSIONS: This patient had a mitochondrial functional deficit consistent with his 9957 mutation. It seems quite likely that this mutation may be responsible for optic nerve and brain injuries. PMID- 15823924 TI - Multiple congenital malformations including severe eye anomalies and abnormal cerebellar development with Dandy-Walker malformation in a girl with partial trisomy 3q. AB - PURPOSE: Ocular anomalies have been associated with numerous chromosomal abnormalities. This report describes partial trisomy 3q in a two-month-old girl with dysmorphic features of the Dup3q phenotype and severe eye and cerebellar malformations. METHODS: Clinical examination and chromosomal analysis were conducted. RESULTS: The karyotype of the propositus was 46,XX, ins(3)(pter --> p25::q27 --> q21::p25 --> qter). She had an abnormal head shape, low-set malformed ears, coarse facies, short webbed neck, abnormal foot position, polycystic kidney, and spina bifida. There was also bilateral microphthalmia that was more severe on the right side, microcornea, and corneal opacity. She had posterior fossa abnormalities, including cerebellar vermis hypoplasia suggestive of a Dandy-Walker (DW) malformation. CONCLUSIONS: This girl with an intrachromosomal duplication of distal 3q and typical phenotype belongs to the severe end of the spectrum for such cases. The ocular manifestations of the 3q duplication syndrome provide additional evidence of the involvement of developmental eye genes in this chromosomal segment. PMID- 15823925 TI - Chromosomal abnormalities and glaucoma: a case of congenital glaucoma with trisomy 8q22-qter/ monosomy 9p23-pter. AB - PURPOSE: To present a case of congenital glaucoma with an unbalanced translocation trisomy 8q22-qter/monosomy 9p23-pter, resulting in trisomy of the GLC1D locus. To perform a literature review of chromosomal abnormalities associated with glaucoma. METHOD: A case report of a family with balanced translocation without glaucoma and unbalanced translocation with congenital glaucoma. PubMed and OMIM databases were searched for reports of chromosomal abnormalities and glaucoma. RESULTS: Other case reports of congenital glaucoma with chromosomal abnormalities in this region were identified. A review of cytogenetics in southeastern Australia found nine cases involving the loss of 9p23 and 10 cases involving mosaicism for trisomy 8, but none had congenital glaucoma. A review of the literature identified reports of glaucoma and chromosomal abnormalities in regions with glaucoma loci mapped by conventional linkage analysis. These include the loci GLC1B, GLC1C, GLC1D, GLC1F, GPDS1, and RIEG2. CONCLUSION: The study of patients with glaucoma and chromosomal abnormalities may help to identify new glaucoma genes. Ophthalmologists can assist with this by requesting cytogenetic studies on congenital and developmental glaucoma cases and interacting with ophthalmic genetics researchers. PMID- 15823927 TI - Hippocampal cytochrome oxidase activity of rats in easy and difficult visual discrimination learning. AB - The present study investigated the functional involvement of the rat hippocampal formation in easy and difficult visual discrimination learning by measuring regional brain cytochrome oxidase (C.O.) activity, an index of neuronal metabolic activity. The results showed that learning was related to C.O. activity in the CA field and the dentate gyrus of the hippocampal formation of rats in the difficult discrimination, whereas learning was related to C.O. activity only in the dentate gyrus of rats in the easy discrimination. The results suggest that difficult visual discrimination learning required greater involvement of the hippocampal formation than easy visual discrimination learning. PMID- 15823928 TI - Correlations between nonverbal intelligence and nerve conduction velocities in right-handed male and female subjects. AB - A neurological theory of intelligence suggesting a direct correlation between nerve conduction velocity and psychometric intelligence was tested. Cattell's Culture Fair Intelligence Test was used to asses the nonverbal intelligence (IQ) of subjects. The motor median nerve conduction velocity from right hand of males was positively correlated with IQ. In subjects with no familial sinistrality (FS ), the motor ulnar-nerve conduction velocity from the right and left hands of males negatively correlated with IQ; there were inverse correlations between IQ and nerve conduction velocity (motor median nerve from right, sensory median nerve from right and left) in females. In subjects with familial sinistrality (FS+), IQ directly correlated with nerve conduction velocity from motor median (right and left), sensory median (right), and motor ulnar (right) nerves, but only in males. The speed hypothesis and neurological theory of intelligence were not supported by these results, which, in contrast, emphasized the importance of sex and familial sinistrality in any theory of intelligence. PMID- 15823929 TI - Complex magnetic fields enable static magnetic field cue use for rats in radial maze tasks. AB - Male Wistar rats were trained in an eight-arm radial maze task (two sessions per day, delayed-non-matching-to-sample) that included an intramaze static magnetic field "cue" (185 microT) specific to the entrance point of one of the arms. Rats were exposed daily for 60 min to a complex magnetic field waveform (theta-burst pattern, 200-500 nT), presented with several different interstimulus intervals (ISIs), either immediately following training sessions or immediately preceding testing sessions. Application of the theta-burst stimulus with a 4000 ms ISI significantly improved the rats' memory for the arm of the radial maze whose position was indicated by the presence of a static magnetic field cue. Reference memory errors were homogeneously distributed among all eight arms of the maze for sham-exposed rats, and among the other seven arms of the maze for complex magnetic field-treated rats. These results suggest that static magnetic field cues may be salient orienting cues even in a microenvironment such as a radial maze, but their use as a cue during maze learning in rats is dependent on whole body application of a specific time-varying complex magnetic field. PMID- 15823930 TI - Self-rated executive function: development of the executive function index. AB - There are several self-rating executive function (SREF) measures in existence that were developed solely in clinical populations or which sample a limited range of executive functions. The Executive Function Index (EFI) was developed here in a normal population with five subscales derived through factor analysis: Motivational Drive, Strategic Planning, Organization, Impulse Control, and Empathy. The content of three second order factors is consistent with the functions mediated by dorsolateral, orbitofrontal, and medial prefrontal circuits. Intrascale reliability and demographic relationships are reported as well as strong correlations with other SREF measures validated in clinical and neuroimaging studies. This brief measure provides a quick and efficient means of collecting data in large samples in order to test hypotheses regarding the role of prefrontal systems in various aspects of behavior and to corroborate findings of other methods, such as objective tests and functional neuroimaging. PMID- 15823931 TI - Specific circular organization of the neurons of human interthalamic adhesion and of periventricular thalamic region. AB - Interthalamic adhesion between the medial surfaces of the left and right thalamus is a variable structure and contains the midline thalamic nuclei, which are not much developed in humans. The research has been done on 6 human brains obtained during routine autopsy (age 45 to 65; 4 male and 2 female). Every tenth 10 microm thick frontal section was stained according to Kluver-Barrera method. In all cases the authors found a specific organization of certain groups of neurons within the interthalamic adhesion (IA) in form of circles on frontal sections. These circular groups were present on all sections but only 1-2 in each. The larger mean diameter of these circular arrangements was R = 229.4 microm, and smaller was r = 203.1 microm. These circular groups within the human IA were formed in average by 7.29 neurons. In periventricular region (PVR) of thalamus similar circular groups of neurons also were present in all cases as in IA. These neuronal groups in PVR were of smaller size than in the IA, with larger mean diameter R = 201.4, smaller mean diameter r = 181.2 microm and they contained fewer neurons, 6.69 on average. All three values (both diameters of circular arrangements, and number of neurons forming them) were significantly smaller in PVR (p < .01). Morphological types and sizes of neurons in both investigated structures (IA and PV) were not different. The circular neuronal groups in IA were formed in 61% of fusiform neurons and in PVR in 48% of fusiform neurons. According to their subependymal localization, size and form, these circular groups can represent in vivo correlates of neurospheres. PMID- 15823932 TI - Neural correlates of visuo-spatial attention during an antisaccade task in schizophrenia: an ERP study. AB - The authors investigated the role of visuo-spatial attention in an antisaccade task using event-related potentials (ERPs) in schizophrenia patients compared to healthy controls. ERPs between 80-130 ms (P100) after stimulus onset showed differences between pro- and antisaccades only for controls and can be related to the suppression of irrelevant stimulus features. Between 150-180 ms (N100), a larger amplitude for anti-compared to prosaccades over centroparietal electrodes showed that processes of visuo-spatial attention seem to be engaged in performance of the antisaccade task. Left temporo-occipitally, this activity was only evident in schizophrenia patients, possibly reflecting additional neuronal recruitment in order to perform the antisaccade task successfully. PMID- 15823933 TI - Discrimination of healthy and brain-damaged elderly using the WAIS-R. AB - The authors evaluated the ability of WAIS-R variables to discriminate normal from brain-damaged persons 75 years of age and older. Fifty-four brain-damaged patients and 54 healthy elderly were matched on the variables of age, education, sex, and race. Significant level of performance differences emerged between the groups on all subtests and the three IQs. No group differences were found on the VIQ-PIQ discrepancy score or range of subtest scatter. Step-wise discriminant analysis revealed that Digit Symbol was the most potent WAIS-R variable, accounting for 34% of the variance and correctly classifying 79.6% of the subjects. PMID- 15823934 TI - Hand preference in Turkish population. AB - The distribution of hand preference (self report) was studied in siblings and their parents originating from all parts of Turkey (N = 22,461). In total sample and siblings, there were significantly more right-handed women than men, and significantly more left-handed men than women--no significant sex difference for parents' handedness. The relative number for the right-handed parents significantly exceeded that for the right-handed siblings; the relative number for the left-handed siblings significantly exceeded that for the left-handed parents. It was concluded that there may be a sex difference in hand preference, but being only about 1% more left-handed men, and only about 1% more right-handed women; the right-handedness in new generation (siblings) is less than that in old generation (parents), due to freeing from cultural pressures against the left hand use in everyday activities. PMID- 15823935 TI - Reduced litter sizes following 48-h of prenatal exposure to 5 nT to 10 nT,0.5 Hz magnetic fields: implications for sudden infant deaths. AB - To test the hypothesis that a narrow intensity band of geomagnetic activity contributes to Sudden Infant Death, 32 pregnant rats were exposed for two to three days before expected parturition either to a coil that generated 0.5 Hz sine-wave, 5 to 10 nanoTesla magnetic fields, or to a reference coil (<1 nT) in the same room. The field was off for 30 min every 4 h during the exposure. The orientation of the coils was perpendicular in space and activated alternately in four blocks of experiments. The litters born to the exposed mothers contained significantly fewer pups (M = 14.1,SD= 2.1) than those exposed to the control conditions (M = 16.2, SD = 2.7). There were significantly fewer numbers of males and fewer numbers of females in litters exposed to the fields generated in the east-west and north-south directions, respectively. These results support the hypothesis that a specific temporal configuration of brief periods of geomagnetic activity can produce an increased incidence of nonvital fetuses, neonates, or infants. PMID- 15823936 TI - Evaluation of brainstem auditory potential in brucellosis patients with and without neurological involvement. AB - This study investigated the changes in brainstem auditory evoked potentials (BAEPs) in acute phase of brucellosis. Twenty-two patients with brucellosis without neurologic involvement and seven patients with neurobrucellosis were included. BAEPs were evaluated before treatment. No patient had a symptom of hearing loss. Patients who had neurologic involvement did not show any abnormalities in their BAEPs recording. This study suggests that patients with brucellosis who are in the acute phase of the disease with or without neurological involvement may have normal BAEPs. BAEPs do not seem to be a sensitive method for central nervous system involvement of brucella patients in the acute phase of the disease. PMID- 15823937 TI - Conduction blocks of lidocaine on crushed rat sciatic nerve: an in-vitro study. AB - The effects of lidocaine on the action potential of crushed sciatic nerves were investigated. The sciatic nerves of the rats were removed 5, 15, 25, and 38 days after the crushing and the compound action potentials were recorded with sucrose gap technique. The nerves were treated with 1 mM lidocaine and the conduction blocks of nonfrequency dependent block (NFDB) and frequency depended block (FDB) at 10, 40, and 100 Hz were determined. In intact nerves, the NFDB effect of lidocaine was 38.4 +/- 0.7 %. On the 15th day after the crush, the NFDB was increased to 60.1 +/- 1.3%. On the 38th day NFDB was decreased to 46.0 +/- 0.8%. The following days after the crushing, lidocaine caused a very high rate of FDB with 10, 40, and 100 Hz stimulation. The high NFDB ratios approached normal levels with the improvement of regeneration, but FDB ratios continued to stay at high levels. The results showed that the ratios of FDB and NFDB were increased on the crushed nerve. It was concluded that, in the regeneration conditions of crushed rat sciatic nerves, the sensitivity to local anesthetic increases more than in intact nerves. PMID- 15823938 TI - Intersession reliability of a protocol to assess reflex activation history in the vastus medialis. AB - The purpose of this study was to determine the reliability of a protocol to assess reflex activation history in the vastus medialis. Eight subjects reported to the laboratory on two occasions. Reflex activation history was assessed by delivering two stimuli of the same intensity 80 ms apart. The dependent variable evaluated was the percentage of the unconditioned reflex amplitude. Eight trials were elicited and averaged on each day. An intraclass correlation coefficient (ICC 2,1) was used to estimate intersession reliability. The ICC for the protocol was found to be 0.9647. The results of this investigation indicate that this technique can reliably estimate reflex activation history in the vastus medialis. PMID- 15823940 TI - Hormone replacement use and cardiovascular function and structure in postmenopausal women. PMID- 15823941 TI - Angiotensin-receptor blockers and diuretics--advantages of combination. AB - Surveys have shown that in as many as half of patients treated for hypertension, blood pressure (BP) is not controlled to target levels; many more persons have undertreated hypertension. Uncontrolled hypertension is a serious risk factor for cardiovascular events such as stroke, heart failure, myocardial infarction and target-organ diseases. Studies have shown that strict BP control significantly reduces the occurrence of these cardiovascular outcomes, in the majority of patients, effective BP control requires two or more antihypertensive agents. The combination of an angiotensin-receptor blocker (ARB) and a thiazide diuretic is appealing, since these agents have complimentary effects on BP reduction, left ventricular hypertrophy and progression of renal diseases. In addition, this combination provide excellent tolerability. The combination of an ARB and a thiazide diuretic may be of particular value in patient populations who tend to have poor BP control on monotherapy, or have additional cardiovascular or renal risk factors. PMID- 15823942 TI - Hormone replacement use, arterial distensibility, cardiac structure and circadian blood pressure profile in menopausal women. AB - Observational and interventional studies that evaluate the impact of hormone replacement therapy (HRT) on cardiovascular changes have produced inconsistent and inconclusive results. The present study sought to elucidate the influence of HRT on aortic distensibility, left ventricular mass (LVM) and 24-h blood pressure (BP) profile in 38 menopausal women who were either HRT users or non-users. The two groups were similar for age, ambulatory BP, aortic distensibility, cardiac mass, lipid profile and body mass index but differed in clinic diastolic BP (DBP). HRT non-dippers had significantly lower clinic and daytime DBP and a smaller nocturnal BP reduction than dippers. Daytime DBP was significantly and inversely related to duration of HRT use. The present study demonstrates that hormonal therapy after menopause lowers DBP, but shows no significant influence on aortic distensibility, cardiac mass or 24-h BP profile. HRT users who were dippers demonstrated a significantly greater nocturnal BP reduction. Long-term controlled trails are needed to better define the effects of estrogen and progestin on the aorta, the heart and 24-h BP profile in normotensive and hypertensive menopausal women. PMID- 15823943 TI - Tumor necrosis factor and interleukin-6 levels in hypertensive patients with and without left ventricular hypertrophy. AB - The objective of this prospective study was to examine the association between serum levels of TNF (tumor-necrosis factor) and IL-6 (interleukin-6) and left ventricular mass in hypertensive patients. Hypertensive patients currently receiving medical therapy were eligible. All subjects underwent echocardiography with measurements of left ventricular (LV) mass and ejection fraction (EF) and had serum levels of TNF and IL-6 measured by ELISA immunoassay. 35 subjects (20F, 15M; mean age 56.4 +/- 10.5 yrs) were studied. 19 patients (54%) had elevated LV mass. Of these patients, 6 (32%) had detectable serum TNF levels and 1 (7%) had detectable IL-6 levels, (p = NS). Hypertensive patients with elevated LV mass do not consistently exhibit elevated cytokine levels when compared to those with normal LV mass. PMID- 15823944 TI - Prevalence and correlates of advanced retinopathy in a large selected hypertensive population. The Evaluation of Target Organ Damage in Hypertension (ETODH) study. AB - OBJECTIVE: To describe the prevalence of advanced retinal microvascular lesions and their associations with cardiac and extracardiac signs of target organ damage (TOD) in a large selected hypertensive population. METHODS: A total of 2172 non diabetic untreated and treated uncomplicated essential hypertensives consecutively attending for the first time our hospital outpatient hypertension clinic and included in the Evaluation of Target Organ Damage in Hypertension (ETODH), an observational ongoing registry of hypertension-related TOD, were considered for this analysis. Advanced hypertensive retinopathy was defined by the presence of any of the following lesions: flame-shaped haemorrhages, soft exudates or cotton wool spots and papilloedema. Left ventricular hypertrophy (LVH), carotid structural abnormalities, such as plaques and intima media (IM) thickening, and microalbuminuria were diagnosed according to the 2003 ESH/ESC guidelines criteria. RESULTS: Among the whole study population, 33 patients (1.5%) were found to have advanced hypertensive retinopathy. Patients with these retinal lesions were similar to those without for age, body mas index, known duration of hypertension, smoking habit, total serum cholesterol, fasting blood pressure and prevalence of antihypertensive treatment; whereas mean systolic and diastolic blood pressures were higher in the former group. The prevalence rates of LVH, carotid plaques, carotid IM thickening and microalbuminuria in patients with and without retinopathy were 57%, 67%, 69%, 19% and 25%, 47%, 44%, 12%, respectively. In a multivariate logistic regression analysis, advanced retinopathy was significantly associated with LVH (OR = 4.0), carotid IM thickening (OR = 2.9), carotid plaques (OR = 2.8), but not with microalbuminuria. CONCLUSIONS: Our study indicated that: (i) advanced retinopathy is a rare finding in non-diabetic hypertensive patients seen in a specialist setting; (ii) a strong relation exists between retinal microvascular lesions and cardiac and macrovascular markers of TOD. PMID- 15823945 TI - The Study on COgnition and Prognosis in the Elderly (SCOPE) - major CV events and stroke in subgroups of patients. AB - The Study on COgnition and Prognosis in the Elderly (SCOPE) assessed the effect of candesartan on cardiovascular outcomes in elderly patients with mild to moderate hypertension. Patients were randomized to candesartan 8-16 mg daily (n = 2477) or placebo (n =2460). Due to extensive add-on therapy, blood pressure reduction was only about 3/2 mmHg greater in the candesartan group than in the control group. Nevertheless, non-fatal stroke was reduced by 28% (p = 0.04) in the candesartan group compared to the control group, and there was a non significant 11% reduction in major cardiovascular events (p = 0.19). This report provides results in pre-specified subgroups of patients (age, gender, diabetes, history of stroke, smoking and cardiovascular risk at randomization). Reductions in major cardiovascular events and stroke with candesartan-based therapy were indicated in all subgroups. A significant interaction between treatment and subgroups was found for one pair of subgroups only; the reduction in major cardiovascular events with candesartan was greater in patients with a previous stroke (64% reduction, p = 0.004) than in those without (5% reduction, p > 0.20). In conclusion, this analysis indicated consistent favourable effects of candesartan-based therapy on major cardiovascular events and stroke across the different subgroups of patients. However, the benefit was particularly pronounced in patients who entered the study with a previous stroke. PMID- 15823946 TI - Hormone replacement therapy and arterial blood pressure in postmenopausal women with hypertension. AB - BACKGROUND: Data on the effect of hormone replacement therapy (HRT) on blood pressure (BP) in hypertensive menopausal women are limited. OBJECTIVE: To investigate the association between HRT and longitudinal change in BP in hypertensive menopausal women. PATIENTS AND METHODS: We recruited a total of 161 hypertensive menopausal women (mean age = 52.2 +/- 6.6 years) attending the hypertension clinic in our hospital that requires HRT to attenuate the effect of menopause symptoms. These women were followed for up tp 36 months, being evaluated every 6 months with measurements of their BP, weight and the number of drugs needed to control their BP. We also measured serum cholesterol levels before and after the initiation of HRT. RESULTS: The systolic BP remained unaffected throughout the whole follow-up period, whereas the diastolic BP was slightly reduced at 6, 24 and 36 months. This decrease was accompanied by an increased need for antihypertensive medication throughout the entire follow-up period, while the body weight also increased at 18, 24, and 36 months. No particular differences were noted with respect to ethnicity, history of pre eclampsia or surgical menopause, before and after the initiation of HRT. Serum cholesterol levels remained unchanged during the evaluation period. Oestrogen progestogen combination therapy use was associated with a lower diastolic BP and a smaller number of antihypertensive drugs compared to other forms of HRT. CONCLUSION: HRT use does not have an adverse gross effect on BP in hypertensive menopausal women who need it, although there may be an increased need for antihypertensive therapy during the 36-moth follow-up period of our study. PMID- 15823947 TI - Large artery stiffness and pulse wave reflection: results of a population-based study. AB - OBJECTIVE: To assess the determination of large artery stiffness and pulse wave reflection in a population sample. METHODS: A 1% random population sample aged 25 65 years was selected in nine districts of the Czech Republic for a survey off cardiovascular risk factors (Czech post-MONICA). Of 891 individual screened in the Pilsen centre in the year 2000, arterial properties were studied in 291 (143 males and 148 females) using the Sphygmocor device. Pulse wave velocity (PWV) in the aorta and in the lowe limbs was measured to assess large artery stiffness. Wave reflection was assessed from radial pulse wave analysis; the main estimated parameter was peripheral augmentation index (PAI) defined as P2/P1 = ratio of pulse pressures measures at the peaks of secondary and primary waves. RESULTS: Aortic PWV increased with age (p < 0.001) and was similar in both sexes. Lower extremity PWV increased with age in women, but not in mean, and its mean value was higher in men (p < 0.001). PAI was higher in females in all age groups (p < 0.001) and increased steeply with age in both sexes (p < 0.001). PAI was increased in current smokers (p < 0.001 in both sexes) and in male smokers, the reflected wave returned earlier than in male non-smokers (p < 0.05). Correlation coefficient of PAI with aortic PWV was 0.22 (p < 0.01), and with central augmentation index (CAI), derived from PAI by mathematical transformation, was 0.94 (p < 0.001). Multiple regression analyses, where age, sex, systolic blood pressure (SBP), total cholesterol level, smoking, glucose level and body mass index were included as independent variables, were performed. PAI was better predicted than aortic or lower extremity PWV is these models (41%, 14% and 10% of variance explained, respectively). Age, female sex, smoking, SBP and total cholesterol predicted PAI level whereas age, SBP and glucose level were the main determinants pf aortic PWV. CONCLUSION: Of the studied arterial parameters, PAI showed the closest association with cardiovascular risk factors. The correlation between PAI and aortic PWV was loose, and both parameters had practically different determinants. PAI, which is obtained by direct measurement above radial artery, was practically identical with the mathematically derived CAI in the studied population sample, and therefore, it is a suitable parameter for studying the phenomenon of wave reflection. PMID- 15823948 TI - Association of periodontitis with increased white blood cell count and blood pressure. AB - This study was aimed to examine the association of periodontitis with white blood cell (WBC) count and blood pressure (BP). In 2002, 424 subjects (manufacturing workers) were investigated for periodontitis by a general dentist. All were Japanese. Among them, 364 subjects (269 men and 95 women) who also attended the next year's (2003) screening were enrolled for this study. Of the 364 subjects, 55 (15.1%) had periodontitis. We also measured the BP and WBC count in periodontitis and non-periodontitis subjects at baseline and 1-year later follow up. The WBC count higher in subjects with periodontitis than in subjects without periodontitis, both at baseline [mean +/- standard error (SE) 6.6 x 10(3) +/- 0.2 x 10(3)/ml vs 5.8 +/- 0.3 x 10(3)/ml; p < 0.001] and follow-up (7.0 +/- 0.3(3)/ml vs 6.5 +/- 0.1(3)/ml; p = 0.003). The systolic BP was higher in subjects with periodontitis than in subjects without periodontitis, both at the baseline (128 +/- 2.1 mmHg vs 120.8 +/- 0.8 mmHg; p < 0.001) and follow-up (129.2 +/- 2.3 mmHg vs 123.0 +/- 0.8 mmHg; p = 0.011), and so was the diastolic BP both at baseline (76 +/- 1.5 mmHg vs 71.2 +/- 0.6 mmHg; p = 0.003) and follow-up (80.5 +/- 1.7 mmHg vs 75.4 +/- 0.7 mmHg; p = 0.004). Periodontitis is associated with increased BP and WBC count. This finding may provide one underlying pathway linking periodontitis and cardiovascular disease. PMID- 15823952 TI - Language disorder: a functional linguistic perspective. AB - This paper explores the issues involved in the linguistic characterisation of disordered discourse and the ways in which a Systemic Functional Linguistic framework addresses these issues. For many years, language disorders were described in terms of formal grammars, with "breakdown" discussed in terms of one or more of the traditional levels of language, i.e., phonology, syntax, and semantics. While it was acknowledged that an individual could have difficulty at one or more of these levels, each was viewed quite separately, with semantics viewed largely from a referential perspective. More recent approaches using functional grammar have broadened this view of language and have provided a model of language that re-conceptualizes the notion of meaning and embraces context as integral to its organisation. Such a model has introduced a different perspective on language into clinical fields, and has enabled researchers and clinicians to explore the skills of speakers with language disorders across a variety of situations and contextual variables, examining the linguistic resources still available to them. This paper introduces principles involved in a functional framework and provides an overview of how these principles have been applied to language disorders to date. In addition, the notion of "disorder" itself is discussed as it is situated in this alternative model. PMID- 15823953 TI - Towards an analysis of dysfunctional grammar. AB - This article applies Systemic Functional Linguistics (SFL) to the study of language dysfunction. It demonstrates the potential that Systemic Functional analysis can offer to one aspect of the analysis of language dysfunction--the failure to realise complexes of clauses. For the purpose of analysis, new concepts and new measures have been created. The newly defined concepts and measures are illustrated in the discourse data of normally developing children and adolescents with brain injury. PMID- 15823954 TI - Theme analysis of narratives produced by children with and without Specific Language Impairment. AB - Theme is defined within the Systemic-Functional Linguistics framework as the point of the departure for the clause and therefore important for text organisation. Aspects of theme are examined in the elicited narratives and story retells from a group of 25 5-8 year-old children with Specific Language Impairment (SLI) and a group of 25 age-matched children with Typically Developing Language. The analysis indicated that subjects with SLI used less variety and complexity in themes with more than one element but this is suggested to reflect localised lexicogrammatical problems. Analysis of the aspects of theme which are more reliant on text-level resources, namely marked theme and theme progression patterns, yielded equivocal results. No significant inter-group difference was found in the subjects' representation of marked Theme and only one of the two progression patterns showed a significant difference. These findings are discussed in relation to the theoretical implications for the description of narrative abilities in SLI. PMID- 15823955 TI - A preliminary investigation of the impact of stuttering on language use. AB - This paper reflects on the application of Systemic Functional Linguistics (SFL) to the field of stuttering. It is argued that the SFL theory may offer insight into the impact that stuttering has on language use. Two case studies are presented to illustrate the application of SFL theory. The two SFL analyses found to be of most use in this pilot study were the analysis of how language is modulated in response to interpersonal aspects of communication (modality) and the analysis of how information is foregrounded (theme). The implications of applying the SFL model to consideration of the utilisation of language resources in people who stutter are discussed. PMID- 15823956 TI - Linguistic resources of individuals with Asperger syndrome. AB - In this paper the available literature on the communication skills of individuals with Asperger Syndrome (AS) is reviewed. Current issues regarding diagnosing and understanding disorders with a core social deficit are examined as well as the potential application of sociolinguistic analyses to the study of Asperger Syndrome, such as that provided from within a Systemic Functional Linguistic framework. The need for investigations that address a broader view of "language" and its use in social situations by individuals with AS as opposed to their mastery of syntactic aspects of language, will be discussed. PMID- 15823957 TI - Some evidence for distinctive language use by children with Attention Deficit Hyperactivity Disorder. AB - This paper reports early findings from a wider study that sought to test the hypothesis that differences in language use exist between children who have a diagnosis of Attention Deficit Hyperactivity Disorder (ADHD) and their non-ADHD peers. Twenty-two, 8 to 12 year-old children (11 with a diagnosis of ADHD and 11 matched peers) comprised the community-based sample. There were nine boys and two girls in each group. All the children were ranked at least average for their language ability on standardised language testing. This case-control study used an approach derived from Systemic Functional Linguistics (SFL) to examine and then compare spoken and written elicited language samples. SFL was thought to be particularly suitable because of its potential to richly describe language in use while taking contextual variation into account. Statistically significant overall differences between the subject and the control groups were found for the measure of grammatical intricacy (GI). Further analyses revealed these differences to be largely attributed to differences between group scores for written material. Discussion explores the potential relevance of these findings to the clinical evaluation of language ability in young school-aged children who have been diagnosed with ADHD. PMID- 15823958 TI - Written discourse and acquired brain impairment: evaluation of structural and semantic features of personal letters from a Systemic Functional Linguistic perspective. AB - This qualitative study investigated written discourse in the form of personal letters written by ten people with aphasia following stroke and ten people with cognitive-language disorder as a consequence of traumatic brain injury, and compared their performance with 15 non brain-damaged writers. Personal letters perform the dual function of providing information and maintaining social relationships. Using the Systemic Functional Linguistics framework for investigation, letters were examined in terms of their dual functions, and at two different strata of language--generic structure and semantic organisation. A small quantum of research suggests that the dissociation between different strata of language (i.e., macro and micro linguistic abilities), identified in the spoken discourse of people with aphasia and people with cognitive-language disorder is mirrored in written discourse. Aphasic writers largely maintain coherent text structure while writers with cognitive-language impairment demonstrate problems with global text coherence and the episodic structure of texts. Results of the generic structure analysis did not support the hypothesis. However, the semantic Move analysis revealed how diminished linguistic resources, most evident in the letters written by the subjects with aphasia, impacted upon the semantic diversity of the text, as well as the interpersonal function of the personal letter. Variable performance as a feature pathology and normality is highlighted and clinical implications discussed. PMID- 15823959 TI - An intracellular delivery vehicle for protein transduction of micro-dystrophin. AB - The Fv fragment of an antibody that selectively targets and penetrates skeletal muscle in vivo was produced as a fusion protein with a micro-dystrophin for use as a delivery vehicle to transport micro-dystrophin into muscle cells. Fv-micro dystrophin was produced as a secreted protein by transient transfection of Fv micro-dystrophin cDNA in COS-7 cells and as a non-secreted protein by permanent transfection in Pichia pastoris. Isolated Fv-micro-dystrophin was shown to be full-length by Western blot analysis. Fv-micro-dystrophin penetrated multiple cell lines in vitro, and it localized to the plasma membrane of a cell line with membrane beta-dystroglycan. In the absence of membrane beta-dystroglycan, it localized to the cytoplasm. Antibody-mediated transduction of micro-dystrophin into muscle cells is a potential therapy for dystrophin-deficient muscular dystrophies. PMID- 15823960 TI - Cell-surface display of E7 antigen from human papillomavirus type-16 in Lactococcus lactis and in Lactobacillus plantarum using a new cell-wall anchor from lactobacilli. AB - The human papillomavirus type-16 (HPV-16) E7 protein is considered a major viral oncoprotein involved in cervical cancer (CxCa) and a potential candidate for the development of a vaccine against this neoplasia. Here, two lactic acid bacteria (the model one Lactococcus lactis and a probiotic one Lactobacillus plantarum) were engineered to deliver an E7 mutant protein (E7mm), which has a reduced transforming activity and consequently, could fit better to therapeutic use in humans than the native form of E7. An efficient cell-surface display of E7mm was obtained in L. lactis using an expression cassette encoding a precursor composed of (i) the signal peptide and the first 15 amino acids of the mature part of the lactococcal Usp45 protein; (ii) E7mm and (iii) the cell-wall anchor of the Streptococcus pyogenes M6 protein (CWA(M6)). This hybrid precursor was produced but not cell-wall anchored in Lb. plantarum. We thus replaced CWA(M6) by the cell wall anchor of a Lb. plantarum protein which allows an efficient cell-wall anchoring of E7mm in this bacterium. The E7mm production and cell-surface display in both L. lactis and a probiotic bacterium, Lb. plantarum, represent one more step towards the development of a safe and effective treatment against CxCa. PMID- 15823961 TI - Macrophage depletion induced by clodronate-loaded erythrocytes. AB - Given the important role of macrophages in various disorders, the transient and organ specific suppression of their functions may benefit some patients. Until now, liposome-encapsulated bisphosphonate clodronate has been extensively proposed to this end. In this paper, we demonstrate that erythrocytes loaded with clodronate can also be effective in macrophage depletion. Here, clodronate was encapsulated in erythrocytes through hypotonic dialysis, isotonic resealing and reannealing to final concentrations of 4.1 +/- 0.4 and 10.1 +/- 0.8 micromol/ml of human and murine erythrocytes, respectively. The ability of clodronate-loaded erythrocytes to deplete macrophages was evaluated both in vitro and in vivo. In vitro studies on human macrophages showed that a single administration of engineered erythrocytes was able to reduce cell adherence capacity in a time dependent manner, reaching 50 +/- 4% reduction, 13 days post treatment. The administration of loaded erythrocytes to cultures of murine peritoneal macrophages was able to reduce macrophage adhesion 67 +/- 3%, 48 h post treatment. In vivo, the ability of clodronate-loaded erythrocytes to deplete macrophages was evaluated both in Swiss and C57BL/6 mice. Swiss mice received 125 microg of clodronate through erythrocytes and 6 days post treatment 69 +/- 7% reduction in the number of adherent peritoneal macrophages and 75 +/- 5% reduction in number of spleen macrophages were observed. C57BL/6 mice received 220 microg clodronate by RBC and 3 and 8 days post treatment 65 +/- 7% reduction in the number of spleen macrophages and the complete depletion of liver macrophages were obtained. In summary, our results indicate that clodronate selectively targeted to the phagocytic cells by a single administration of engineered erythrocytes is able to deplete macrophages, even if not completely. The transient suppression of macrophage functions through clodronate-loaded erythrocytes can be used in many biomedical phenomena and research applications. PMID- 15823962 TI - Aspirin induces its anti-inflammatory effects through its specific binding to phospholipase A2: crystal structure of the complex formed between phospholipase A2 and aspirin at 1.9 angstroms resolution. AB - Phospholipase A2 is potentially an important target for structure-based rational drug design. In order to determine the involvement of phospholipase A2 in the action of non-steroidal anti-inflammatory drugs (NSAIDs), the crystal structure of the complex formed between phospholipase A2 and aspirin has been determined at 1.9 angstroms resolution. The structure contains 915 protein atoms, 1 calcium ion, 13 atoms of aspirin and 105 water molecules. The observed electron density of the aspirin molecule in the structure was of very high quality thus allowing the precise determination of its atomic coordinates leading to the clear description of its interactions with the enzyme. The structure of the complex clearly shows that aspirin is literally embedded in the hydrophobic environment of PLA2. It is so placed in the substrate binding channel that it forms several important attractive interactions with calcium ion, His 48 and Asp 49. Thus, the structure of the complex clearly shows that aspirin occupies a favourable place in the specific binding site of PLA2. The binding studies have shown that acetyl salicylate (aspirin) binds to PLA2 enzyme specifically with a dissociation constant of 6.4 x 10(-6) M. The structural details and binding data suggest that the inhibition of PLA2 by aspirin is of pharmacological PMID- 15823963 TI - Targeted gene delivery to hepatoma cells using galactosylated liposome-polycation DNA complexes (LPD). AB - A major goal for gene therapy is to obtain targeted vectors that transfer genes efficiently to specific cell types. The liver possesses a variety of characteristics that make this organ very attractive for gene therapy. In the present study, four cholesterylated thiogalactosides 1a approximately d with different spacer length were synthesized to formulate novel lipid-polycation-DNA (LPD) complexes, which were composed of galactosylated cationic liposomes, protamine sulfate and plasmid DNA. The galactosylated LPD1c significantly improved the levels of gene expression in cultured hepatoma cells HepG2 and SMMC 7721, while LPD1a and LPD1b did not significantly improve the levels compared with non-galactosylated LPD. Meanwhile, increased transfection activity was not observed in mouse fibroblasts L929 for galactosylated LPDs. Cytotoxicity of galactosylated LPDs assay showed they had no obvious toxicities to L929 cells and HepG2 cells. In summary, the length of the spacer between the anchor and galactose residues was important for the recognition of asialoglycoprotein receptor. The LPD1c described here, combining the condensing effect of protamine and the targeting capability of cholesterylated thiogalactosides, are potentially useful gene carriers to liver parenchymal cells. PMID- 15823964 TI - Improvement of the inhibitory effect of xanthones on NO production by encapsulation in PLGA nanocapsules. AB - For the first time the inhibitory effect of xanthone and 3-methoxyxanthone on nitric oxide (NO) production by IFN-gamma/LPS activated J774 macrophage cell line is reported. A remarkable improvement of this effect promoted by encapsulation of these compounds in nanocapsules of poly (DL-lactide-co-glycolide) (PLGA) is also demonstrated. A weak inhibitory effect of 3.6% on NO production by activated macrophages was observed for xanthone at the highest studied concentration (100 microM). This effect was slightly higher for 3-methoxyxanthone at the same concentration, producing a reduction of 16.5% on NO production. In contrast, equivalent concentrations of xanthone and 3-methoxyxanthone incorporated in nanocapsules produced a significant decrease on NO production of 91.8 and 80.0%, respectively. Empty nanocapsules also exhibited a slight NO inhibitory activity, which may be due to the presence of soybean lecithin in the composition of the nanosystems. The viability of the macrophages was not affected either by free or nanoencapsulated xanthones. Fluorescence microscopy analysis confirmed that a phagocytic process was involved in the macrophage uptake of xanthone- and 3 methoxyxanthone-loaded PLGA nanocapsules. Phagocytosis might be the main mechanism responsible for the enhancement of the intracellular delivery of both compounds and consequently for the improvement of their biological effect. PMID- 15823965 TI - Lymphatic chemotherapy induces apoptosis in lymph node metastases in a rabbit breast carcinoma model. AB - The purpose of the study was to evaluate the potential of lymphatic chemotherapy in inducing apoptosis in axillary lymph node metastases in a rabbit breast cancer model. A total of 30 female New Zealand rabbits with mammary implantation of VX2 carcinomas were divided into three groups randomly, with ten in each. Treatment was carried out once axillary lymph node reached 5 mm in the maximum diameter. Group A received a subcutaneous injection of liposomal adriamycin (LADR) adjacent to the breast tumor. Group B received free adriamycin (FADR) administered into the auricular vein. Group C received a sham treatment. The dose of adriamycin in each administration was 1 mg/kg in groups A and B. Treatment was repeated every 48 h. Axillary lymph nodes were dissected out 48 h after the third treatment. The nodal sizes before and after the treatment were measured. The therapeutic effect was evaluated in terms of the node volume ratio and apoptotic index (AI) of metastatic cells in nodes identified with TUNEL technique. The significance of difference was determined with one-way ANOVA followed by the Fischer LSD test. Compared to group C, the enlargement of lymph nodes was sufficiently slowed down in both groups A and B, and group A showed a further strong inhibitory effect than group B (P = 0.002). Apparent VX2 cell apoptosis was detected in the lymph nodes of groups A and B. The average AI in group B (15.31%) was significantly higher than in group C (5.16%). The highest AI was found in animals of group A (21.73%), with a further significant difference from group B (P = 0.000). These data suggest that lymphatic chemotherapy appears to be a promising method to induce apoptosis in lymph node metastases PMID- 15823966 TI - Freshwater fishes, dispersal ability, and nonevidence: "gondwana life rafts" to the rescue. PMID- 15823967 TI - Public health epidemiology--what next? PMID- 15823968 TI - Occupation, work environment, and disability pension: a prospective study of construction workers. AB - AIMS: This study investigated the importance of occupation and work environment for the risk of disability pension among construction workers in Sweden. METHODS: The data include around 389,000 construction workers who participated in health examinations from 1971 to 1992. During that period 68,000 participants received disability pension in some form. Between 1985 and 1992 questionnaires were used to collect data regarding hazards in the work environment. Some 87,000 participants answered that questionnaire, and almost 6,000 of these were in receipt of a disability pension. To analyse the risk of disability pension in different occupations, incidence rates (SIR) were calculated according to the person-year method. Logistic regression was used to calculate age-controlled odds ratios according to physical and psychosocial work environment factors. RESULTS: The results show considerable variation in risk for disability pension between different occupations within the construction industry. Furthermore, analysis of work environment factors indicated associations with physical, ergonomic, and psychosocial work environment factors. DISCUSSION: A considerable variation between occupations indicates that characteristics in the work situation are important when explaining the risk of disability pension. The results from this study indicate that improvements in working conditions are an important area of intervention in order to facilitate and prolong labour market participation among elderly people. PMID- 15823969 TI - More physicians in public health: less public health work? AB - AIM: The object was to assess changes in work priorities in local public health medicine in Norway over the period from 1994 to 1999. METHODS: Two cross sectional studies were undertaken of physicians working in local public health medicine in all Norwegian municipalities, using a postal questionnaire. RESULTS: Half of the physicians working in public health in 1999 were recruited after 1994. Although the number of physicians working in public health increased from 505 in 1994 to 555 in 1999 (10%) an estimation of the total weekly hours worked decreased by 3.7% from 8,715 hours in 1994 to 8,386 hours in 1999. The vast majority of physicians worked in combined posts (87%), and they reduced their engagement in public health by 2.6 hours on average from 1994 to 1999. The reduction depended on remuneration model, speciality in community medicine, and municipality size. CONCLUSIONS: Local public health in Norway was under pressure in the 1990s. For public health physicians, preventive medicine lost out to clinical work. No promising signals of change in the professional or political framework or in incentives for public health work are seen. PMID- 15823970 TI - Overweight and obesity among Norwegian schoolchildren: changes from 1993 to 2000. AB - AIM: The aim of the present study was to investigate the prevalence rate of overweight and obesity in a nationwide survey of fourth- and eighth-grade Norwegian schoolchildren, the changes in overweight rates among eighth graders from 1993 to 2000, and to investigate factors associated with overweight. METHODS: Cross-sectional dietary surveys were conducted on nationally representative samples of eighth graders in 1993, and among fourth and eighth graders in 2000. Self-reported weight and height was available from 1,650 eighth graders in 1993, and from 664 fourth and 825 eighth graders in 2000. Data on dietary pattern, social class, and physical activity were also available. RESULTS: In 2000 the prevalence of overweight and obesity among the fourth graders was 18.5% and 3.6%, respectively, and among the eighth graders 11.5% and 1.8%, respectively. A marked increase in overweight and obesity among Norwegian eighth graders from 1993 to 2000 was observed; the prevalence of overweight and obesity increased by 4 and 0.9 percentage points, respectively. Social class, time spent watching TV or in front of a computer, and breakfast frequency were associated with the odds of being overweight. An inverse relation between intake of sweets and overweight was observed. Validation studies demonstrated a high correlation between self-reported and measured height and weight in both age groups. CONCLUSION: The present study demonstrates a high prevalence of overweight and obesity among schoolchildren, especially among fourth graders in Norway. Moreover, the proportion of overweight and obese children has increased greatly among eighth graders during the last decade. PMID- 15823971 TI - Assistant nurses in the Swedish healthcare sector during the 1990s: a hard-hit occupational group with a tough job. AB - AIMS: The aim of the study was to explore hospital-based assistant nurses' experiences of psychosocial "stressors", following a period of substantial layoffs (43%) and ongoing healthcare reorganizations. METHODS: An interview study was carried out with 11 assistant nurses working in the same hospital. The interviews took place in 1997, in connection with the last round of redundancies, and were followed up in 1998 and then in 2001. Interviews were audiotaped and transcribed; the content was then analysed. RESULTS: Two main themes were identified from the women's perceived stressors: (a) a hard-hit occupational group experiencing "energy-consuming adjustments", and a "weak position" at the continuing workplace. Job insecurity meant fear of losing valued work tasks in nursing care (de-skilling). The common feature was the duality in the women's descriptions of feeling qualified in nursing care but being treated like a maid, or having intimate practical knowledge but no formal competence; (b) a tougher but underpaid job including "heavy workload" concurrent with "organizational shortcomings", and "frozen salary trends" with a simultaneous feeling of lacking the power to improve their situation. CONCLUSIONS: Our results underscore the importance of the employer's attention to the remaining workers in connection with downsizing, particularly when the reduction of the workforce has been as dramatic as in this case. It is also important to understand the ongoing dilemma (strain) for the assistant nurses, who are faced with increasing demands for further formal qualifications in hospital care, while maintaining a strong occupational desire to keep their highly valued job working close to the patient. PMID- 15823972 TI - Primary health care and hospital interactions: effects for hospital length of stay. AB - AIMS: Norwegian healthcare services are divided between primary and secondary care providers. A growing problem is that every third patient of 75 years of age or more experiences an extended stay in a somatic hospital while waiting to be sent to primary healthcare services. The interaction between these two levels of healthcare services is analysed to examine the effect on a patient's length of stay in hospital. METHODS: Recent studies have asserted that research on length of stay in hospital should include influential factors such as system variation and system characteristics, in addition to standardizing for case-mix. New organizational routines are identified in 50 Norwegian somatic hospitals. A multivariate linear regression is used in both a static and a dynamic model to explain variations in hospital length of stay and in additional length of stay (5% of stays are defined as outliers). RESULTS: The study shows that newly specialized structures constructed to enhance the interaction between the two levels have had no effect. Length of stay is dependent on the capacity of the primary healthcare provider and on the share of elderly in the hospital catchment area, the type of patients, the procedure performed, and the characteristics of the hospital. CONCLUSION: Variation in length of stay between hospitals is primarily explained by the capacity of primary healthcare providers. However, some support is found in the dynamic model that introduces the proposition that a hospital-owned hotel would decrease the length of stay of patients in hospital. PMID- 15823973 TI - Fatigue in the general population: a translation and test of the psychometric properties of the Norwegian version of the fatigue severity scale. AB - OBJECTIVE: A study was undertaken to test the psychometric properties of the Fatigue Severity Scale (FSS), and to explore the relationship between fatigue and sociodemographic variables in the general population. METHOD: A national representative sample of 1893 respondents was randomly selected from a pool of 4,000 Norwegians aged 19-81 years. A mailed questionnaire that included the FSS was used to measure fatigue severity. RESULTS: The FSS showed satisfactory internal consistency (Cronbach's alpha 0.88). The prevalence of high fatigue (FSS score > = 5) was 23.1% in the total sample. More women (26.2%) than men (19.8%) experienced high fatigue (p = 0.004). Respondents with chronic illness (more than six months) reported a higher mean (M = 4.69, SD = 1.35) than the rest of the sample (M = 3.67, SD = 1.17) (p<0.001). An inverse correlation was found between fatigue and level of formal education (r = 0.20, p<0.001). CONCLUSION: The psychometric properties of the Norwegian version of FSS were satisfactory. To avoid over-diagnosing people for high level of fatigue, the threshold for high fatigue probably should be 5 on the FSS scale instead of 4 as had been suggested originally, but further validation of the cut-off point is needed. PMID- 15823974 TI - Does high income buffer the association between adverse working conditions and ill health? AB - AIM: A study was undertaken to analyse the possible interaction between work environment and income for the probability of self-rated health being less than good. METHODS: Data from the Swedish Survey of Living Conditions for the years 1998 and 1999 were analysed. Employed 20- to 64-year-olds with a registered wage were included (n=5982). The synergy index (SI) was applied, using odds ratios from logistic regressions for men, women, and all. Low and high levels of physical demands, decision authority, skill discretion and psychological demands were separately combined with low- and high-wage income (median split). Full-time work and four sociodemographic factors were controlled for. RESULTS: Significant synergy was found for women when they were exposed to low income and a low level of skill discretion (SI=1.46 [1.01-2.13]), although this was attenuated by education level (SI=1.47 [0.96-2.25]). In general (both sexes), poor health caused by low income and unfavourable work is additive rather than multiplicatively exaggerating the risk among the jointly exposed. CONCLUSION: Work exposures in the form of high physical load, low levels of decision authority and skill discretion, or a high level of psychological demands were significantly related to poor health also when income was high, suggesting that high income does not seem to buffer the detrimental effects of adverse working conditions. As nearly half of employed women were found to be in circumstances marked by synergy, it seems a relevant public health issue to improve these women's conditions at work, by simultaneously increasing, for example, job variety and wages. PMID- 15823975 TI - Acceptance of the smoking cessation intervention in a large population-based study: the Inter99 study. AB - AIMS: Potential exists for improving the impact of quit-smoking programmes, by recruiting smokers in early motivational stages, by using active recruitment strategies, and by offering professional assistance to quit. METHODS: This was a randomized population-based intervention study, in Copenhagen, Denmark. A total of 2,408 daily smokers in all motivational stages were included. All participants completed a questionnaire, and underwent a health examination and a lifestyle consultation. Smokers in the high-intensity intervention were offered assistance to quit smoking in smoking cessation groups. RESULTS: Before the lifestyle consultation only 11% of the smokers stated that they planned to quit within one month. After the lifestyle consultation 27% accepted smoking cessation in groups and an additional 12% planned to quit without assistance. Of the smokers who accepted smoking cessation groups 23% had not been planning to quit before the lifestyle consultation. Being a woman, having high tobacco consumption, having a long smoking history, having tried to quit within the previous year, and having a higher motivation to quit predicted participation in smoking cessation groups. CONCLUSIONS: It was possible to recruit a large number of smokers in early motivational stages by using active recruitment strategies and by offering assistance to quit. Lifestyle consultations markedly increased the number of smokers willing to try to quit. Smokers preferred assistance to quit in a smoking cessation group to quitting on their own; therefore, it is important to improve recruitment strategies for smoking cessation programmes. PMID- 15823976 TI - Willingness to pay for hypothetical malaria vaccines in rural Burkina Faso. AB - AIMS: This study aims to set priorities for anti-disease malaria vaccines by determining community preference in a hyperendemic area. METHODS: A bidding game technique was used to elucidate willingness to pay in rural Burkina Faso and 2,326 adults were interviewed. RESULTS: It is shown that there are significant differences between community preference for an anti-disease vaccine aimed at reducing pathology in pregnant women, and for a vaccine directed against childhood malaria. While the target population was willing to pay CFAfr 2101 for a vaccine against maternal malaria, its members were prepared to pay only CFAfr 1433 for a vaccine against childhood malaria. CONCLUSIONS: Whilst it is increasingly likely that anti-disease malaria vaccines will become available in the foreseeable future, lessons from the past suggest that a lack of acceptance and support from the intended recipients may lead to less than optimal compliance, and hence efficacy. For the planning of vaccine development and application strategies, it is therefore highly important to take community views into account. Here it is argued that such information could help researchers and funding agencies to set priorities for future vaccine research. PMID- 15823977 TI - From healthy survivors to sick survivors--implications for the twenty-first century. AB - BACKGROUND: Hypotheses concerning compression of morbidity have gained support, particularly due to improved lifestyles from 1950 to date, but now the increase is largely due to improvements in healthcare. METHODS: Survey data from Sweden were used to test whether the older population aged 65-84 years during 1995-2002 had more longstanding illnesses than the older population of 1980-87 or 1988-94. RESULTS: There was an increased prevalence of many longstanding illnesses among the elderly in Sweden between 1988-94 and 1995-2002. The increase was especially pronounced among those with at least three longstanding illnesses. For diabetes, heart disease, and hypertension, the prevalence among elderly men increased by over 20%. However, those reporting these kinds of longstanding illnesses perceive improved health and are less restricted in their daily activities. CONCLUSIONS: Results from Sweden indicate that many countries approaching Sweden's life expectancy will have an increased need for care for the oldest population in the future. The compression-of-morbidity hypothesis is being challenged, probably due to improvements in healthcare. PMID- 15823978 TI - Price restrictions and other restrictions on alcohol availability in Denmark and Sweden: a historical perspective with implications for the current debate. AB - BACKGROUND: Current political debate in Sweden is mainly centred on lowering taxes on alcohol in order to "harmonize" prices with those in neighbouring countries, although the evidence of a negative association between prices and alcohol consumption is more than convincing. METHOD: Total per capita consumption figures for twentieth-century Denmark and Sweden are utilized to illustrate the astonishing effects on consumption patterns of active government policies to restrict availability. PMID- 15823979 TI - 22q11.2 deletion syndrome: introduction. PMID- 15823980 TI - 22q11.2 deletion syndrome: genetics, neuroanatomy and cognitive/behavioral features keywords. AB - This paper presents a conceptual review of the genetic underpinnings of 22q11.2 Deletion Syndrome. The neuroanatomical, neuropsychological, behavioral, and psychiatric phenotype associated with 22q11.2 Deletion Syndrome is also explored, including variables that are thought to affect symptom expression. The history of the deletion syndrome is described, and future directions for continued research are discussed. PMID- 15823981 TI - Neuropsychological profile and neuroimaging in patients with 22Q11.2 Deletion Syndrome: a review. AB - 22q11.2 Deletion Syndrome is associated with cognitive, behavioural, and psychiatric problems and is known to affect brain structure. Recently, 22q11.2 Deletion Syndrome has been proposed as a disease model for a genetic subtype of schizophrenia. In this paper we discuss the currently available literature on neurocognitive functioning and brain anatomy in patients with 22q11.2 Deletion Syndrome, and how this contributes to our understanding of the neurobiology of schizophrenia. Research on cognitive functioning in 22q11.2 Deletion Syndrome patients suggests a specific cognitive profile with impairments on arithmetical, visuo-spatial, and executive tasks and relatively preserved language skills. Prominent findings of neuroimaging studies in 22q11.2 Deletion Syndrome patients are: reduction of overall brain volume, midline defects, structural alterations of cerebellum and frontal lobe, white matter abnormalities, and decreased grey matter volumes in parietal and temporal areas. We describe how brain abnormalities in patients with 22q11.2 Deletion Syndrome may contribute to the understanding of the clinical syndrome including cognitive impairments, psychotic symptoms, and social and communication problems. PMID- 15823982 TI - Neuropsychological characteristics of children with the 22q11 Deletion Syndrome: a descriptive analysis. AB - Previous reports of cognitive functioning in children with the 22q11 Deletion Syndrome have reported marked variability in IQ and achievement subtest scores. Studies have begun to explore neuropsychological function in 22q11 DS however results are inconsistent and the profile incomplete. We assessed 40 children ages 5-12 with 22q11 DS. Consistent with past results, visual-spatial memory was significantly lower than verbal memory. Differentially lowered scores were found only in visual attention, working memory and motor function. Contrary with some past results quantitative, verbal ability, and visual spatial memory scores were within 1 SD from the standardization sample mean. Motor behavior, not typically discussed with regard to 22q11 DS school-age children, may be critical to incorporate in neurocognitive studies of children with 22q11 DS. Implications of these findings are considered with regard to past results. PMID- 15823984 TI - Gender-moderated dorsolateral prefrontal reductions in 22q11.2 Deletion Syndrome: implications for risk for schizophrenia. AB - To investigate the impact of the microdeletion on morphology of the prefrontal cortex in 22q11.2 Deletion Syndrome (22q11.2 DS), high-resolution, anatomic magnetic resonance imaging was performed on 19 children and adolescents with 22q11.2 DS (11 females, 8 males) and 18 unaffected controls (10 females, 8 males). Tissue volumes of the dorsolateral, dorsomedial, orbitolateral, and orbitomedial prefrontal cortex were measured. Tasks of executive function and working memory were administered to investigate the association between anatomy and function. Whole brain volume and frontal lobe tissue volume were preserved in girls but reduced in boys with 22q11.2 DS relative to age-matched controls. Dorsolateral prefrontal cortex (DLPFC) volumes were reduced in participants with 22q11.2 DS, although the gender-by-diagnosis effect found for frontal lobe was not as robust for DLPFC. DLPFC volumes were associated with performance on tasks of planning and emotional facial recognition. Longitudinal studies are needed to clarify whether gender differences in frontal lobe and DLPFC persist with development, and whether the volumes of the DLPFC are associated with eventual deterioration in adaptive/psychosocial function that may presage the onset of schizophrenia, for which individuals with 22q11.2 DS are at a disproportionately high risk. PMID- 15823983 TI - Memory and learning in children with 22q11.2 deletion syndrome: evidence for ventral and dorsal stream disruption? AB - This study examined memory functioning in children and adolescents with 22q11.2 Deletion Syndrome (DS; velocardiofacial syndrome). An overall verbal better than nonverbal memory pattern was evident on the Test of Memory and Learning (TOMAL), with children with 22q11.2 DS performing significantly below their siblings and children with low average IQ but similar to children with autism on facial memory. Children with 22q11 DS also performed significantly below their siblings on tests of verbal working memory. Children with autism performed significantly poorer than the siblings of children with 22q11.2 DS only on their recall of stories. Delayed recall was significantly poorer in children with 22q11.2 DS and children with autism, compared to sibling controls. Although there were no significant group differences on tests of multiple trial verbal or visual learning, a relative weakness was noted with multiple trial visual learning in children with 22q11.2 DS and their siblings, suggesting that an alternative or interactive factor other than the deletion may account for the relatively better verbal compared to nonverbal memory abilities. Deficits in facial memory in children with both 22q11.2 DS and autism suggest disruptions in ventral temporal pathways such as between fusiform gyrus and parahippocampal/hippocampal regions whereas deficits in verbal working memory in children with 22q11.2 DS implicates dorsolateral prefrontal regions, both intimating aberrant white matter pathways. PMID- 15823985 TI - Atypical neuropsychological profile in a boy with 22q11.2 Deletion Syndrome. AB - In this article the general and specific cognitive impairments of the boy R.H. with a de novo deletion 22q11.2 are described. His full-scale IQ was 73, and he obtained only slightly better verbal than non-verbal subtest scores. Neuropsychological assessment revealed specific impairments in perceptual categorization of objects presented suboptimal, matching of unfamiliar faces, and verbal learning and memory. In contrast, he performed in accordance with his intelligence level on other visual perceptual tasks, on non-verbal learning and memory tasks, and on attention tasks. Voxel-wise statistical comparison of a high resolution T1-weighted magnetic resonance image of R.H's brain with similar images obtained from 14 normal control children revealed as major abnormalities a reduction of the right inferior parietal and superior occipital lobe, and a bilateral reduction of deep white matter behind the inferior frontal gyrus. These cognitive impairments and MRI abnormalities are not commonly described in 22q11.2 Deletion Syndrome and may indicate a larger heterogeneity in the neurocognitive phenotype than currently evidenced. At least in this boy the microdeletion seems to have interfered with the development and functioning of particular neural subsystems, while the structure and functioning of other subsystems was left intact. PMID- 15823988 TI - An overview of research on increasing indices of happiness of people with severe/profound intellectual and multiple disabilities. AB - PURPOSE: This paper was to provide an overview of research studies aimed at increasing indices of happiness of persons with severe/profound intellectual and multiple disabilities. METHODS: Computerized and manual searches were carried out to identify the studies published from 1990 to 2004 (i.e., the period during which the issues of quality fo life and happiness in people with disabilities have become more prominent). Twenty-four research studies were identified. They involved the use of six different procedures, that is, structured stimulation sessions, microswitch-based simulation sessions, leisure activities and favourite work tasks or conditions, positive environment or positive behaviour support programmes and mindful caregiving, favourite stimulation automatically delivered on exercise engagement, and snoezelen. RESULTS AND CONCLUSION: Data tended to be positive with increases in the participants' indices of happiness, but some failures also occurred. The outcomes were discussed in relation to (a) methodological issues, such as designs of the studies, length of the intervention, and number of participants, and (b) personal and practical implications of the procedures. Some suggestions for future research (particularly focused on extending evidence and overcoming present methodological weakness) were also examined. PMID- 15823989 TI - Health related quality of life after lumbar disc surgery: a prospective study of 145 patients. AB - PURPOSE: This study compared health related quality of life (HRQOL) among lumber disc surgery patients to general population, and assessed the relationships between HRQOL and other outcome measures 2 and 14 months after surgery. METHODS: The 15D instrument was use to measure HRQOL in 145 lumber disk surgery patients. Subjective outcome was also assessed by the Oswestry index, Short Depression Inventory (SDI) and pain, and objective outcome by trunk muscle strength and mobility of the spine. RESULTS: The mean (SD) 15D score among the patients 2 months after surgery was 0.880 (0.104) compared to 0.941 (0.076) among general population (p = 0.016). The 15D, Oswestry and SDI scores remained unchanged during the follow-up, while the improvements in the spine mobility and trunk muscle strength were significant. Back pain decreased by 68% and leg pain by 74% from preoperative level to 2 months check-up. Leg pain further decreased during the following 12 months, while back pain remained unchanged. HRQOL was associated with age, pain, Oswestry and SDI indices, but not with physical function. CONCLUSION: HRQOL measured by 15D was lower in lumber disc surgery patients compared to general population. 15D was associated with the subjective outcome measures of pain, Oswestry and SDI indices, but not with the objective measures of physical function. Thus, a combination of "disease specific" and "generic" measures is recommended to be used to obtain more accurate information about the overall welfare of individual. PMID- 15823990 TI - Claimed walking distance of lower limb amputees. AB - PURPOSE: Walking ability in general and specifically for lower limb amputees is of major importance for social mobility and ADL independence. Walking determines prosthesis prescription. The aim of this study was to mathematically analyse factors influencing claimed walking distance of lower limb amputees of 500 m or more. METHOD: A total of 437 patients returned two questionnaires: the Groningen Questionnaire Problems after Leg Amputation, in which walking distance was assessed, and the RAND 36. RESULTS: The chance of walking 500 m or more reduced when a transfemoral amputation was performed. The chance reduced even more when phantom pain or stump pains were present. If the amputation was performed because of vascular disease or because of vascular problems because of diabetes the chance reduced again. Independently of these factors, age reduced the chance of walking 500 m or more. CONCLUSION: The chance of walking 500 m or more reduces with increase in age and a more proximal amputation. The chance reduces even further when the amputation is performed because of diabetes or vascular disease and also if phantom pain and or stump pain is present. PMID- 15823991 TI - Effects of surface electrical stimulation on the muscle-tendon junction of spastic gastrocnemius in stroke patients. AB - PURPOSE: The purpose of this study was to explore the effects of spasticity suppression by surface electrical stimulation (ES) o the muscle-tendon junction of spastic gastrocnemius muscles in stroke. METHODS: Twenty-four neurologically stable stroke patients (aged 41-69 years, 12-35 months post-stroke), with spasticity graded 2 or 3 on the modified Ashworth scale, were recruited and divided into two groups. In the ES group, each patient received 20 min of surface ES once daily, 6 days per week for 1 month. In the control groups, ES was used with stimulation intensity kept at zero. To evaluate the therapeutic effect, the modified Ashworth scale, Fmax/Mmax ratio, H-reflex latency, H-reflex recovery curve, and the 10-m walking time were tested before and after the 1-month of treatment. RESULTS: In the ES group, the modified Ashworth scale showed a trend toward reduced spasticity after 1 month of treatment. The Fmax/Mmax ratio decreased from 8.10% +/- 4.84% to 4.00% +/- 1.36%; the H-reflex latency increased from 28.87 +/- 2.45 ms to 29.40 +/- 2.57 ms; the H-reflex recovery curves indicated a downward shift; and the 10-metre walking time significantly decreased after ES. In the control group, none of the measures showed a statistically significant change. CONCLUSIONS: In this study, we demonstrated a way to suppress spasticity at a metameric site and to increase walking speed effectively by applying surface ES on the muscle-tendon junction of spastic gastrocnemius muscles. PMID- 15823992 TI - Late multidisciplinary rehabilitation in young people after stroke. AB - PURPOSE: Stroke has a major impact on physical and cognitive ability, and quality of life. This study aims to examine the effect of multidisciplinary rehabilitation on outcome in an Irish young stroke population. METHODS: Fifty consecutive patients younger than 65 were prospectively recruited to this observational study. Physical and cognitive ability, and quality of life were measured on admission and discharge. RESULTS: All patients were transferred to the rehabilitation unit from the acute hospital where they had received their initial management. Median length of time from stroke onset to transfer was 112 days. After a median of 70 days, 43 patients were discharged home. Patients made significant gains in physical (Barthel index 12 to 18) and cognitive ability (mini-mental state examination 22 to 25). Quality of life improved in all eight domains of the 36-items Short Form with four domains returning to level seen in the general population. CONCLUSIONS: This study documents the improvements in physical and cognitive abilities, and quality of life in a sample of patients in rehabilitation. A comprehensive rehabilitation programme that includes acute and late multidisciplinary phases can improve the outcome of patients following stroke. PMID- 15823994 TI - A lump in the throat: Should speech and language therapists treat globus pharyngeus? AB - PURPOSE: The causes of the feeling of a lump in the throat (globus pharyngeus) are disputed. The symptoms are persistent and may lead clients to change their diet and suffer a reduced quality of life. Coexisting swallowing difficulties are often reported but no neurological or structural abnormality can be found. Psychological factors are likely to play a part in the condition and an eclectic approach to therapy appears to be warranted. This paper reports on a treatment programme that includes education, reassurance and the use of exercises. METHOD: Fourteen clients participated. Their progress was assessed using the Glasgow and Edinburgh Throat Scale (GETS). A baseline period was used to assess the stability of their symptoms prior to therapy. This was followed by a group therapy session, use of the exercises and a further reassessment. Videofluoroscopy was conducted before and after therapy. RESULTS: Clients score on the GETS improved. However, change was seen both during the baseline and the treatment making it unclear which aspects of the treatment were effective. Improvement during the baseline suggests that clients benefit from attention and reassurance. It is unclear, therefore, whether the therapy session and the exercises provide a specific benefit or a general benefit due to continued attention. The clients' videofluoroscopies were unchanged after therapy. The presence of residue after the swallow in the valleculae and the pyriform sinuses may contribute to the symptoms of globus. CONCLUSION: The treatment is economical and appears to benefit clients. However, further research is needed to distinguish whether it has a specific effect or whether clients benefit from general attention and reassurance. PMID- 15823993 TI - Depression in adults with disabilities, in primary care. AB - PURPOSE: This research was design to answer the question: Does the prevalence of depression differ between adults with and without disability, in the same family medicine practice? METHOD: A retrospective cohort design was used, to study depression among adults, with and without primary disabling conditions, receiving primary care in either a university based urban or rural family practice setting. RESULTS: When we compared individuals with disability to those without disability, and controlled for individual characteristics, the relative risk for depression was significantly lower for individuals with autism (Relative Risk (RR) 0.20: 95% Confidence Interval (CI) 0.05-0.55), cerebral palsy with mental retardation (RR 0.40: 95% CI: 0.24-0.65), and MR (RR 0.56: 95% CI: 0.39-0.77). The risk for depression was significantly higher for those with cerebral vascular accidents/stroke (RR 2.18: 95% CI: 1.72-3.76) and traumatic brain injury (RR 2.55: 95% CI: 1.72-2.77). The earliest onset of depression was among individuals with traumatic disabilities and milt mental retardation. Our estimate of depression prevalence for the non-disabled and disabled primary care patients was 22.8% and 24.9% respectively, when patients with disabilities were grouped together (p = 0.008). CONCLUSION: It is important for physicians to recognize the higher prevalence of depression among patients with adult onset disabilities (e.g. stroke, traumatic brain injury). In addition, they should be aware of lower prevalence of depression among many individuals with lifelong disabilities, such as mental retardation, cerebral palsy, and autism. PMID- 15823995 TI - Modeling social reintegration in persons with spinal cord injury. AB - PURPOSE: This study was undertaken to identify and develop a model of the factors related to social reintegration in persons with spinal cord injury (SCI). METHOD: A convenience sample of 145 persons with SCI living in two cities in South Korea anonymously completed a questionnaire that comprised the assessment of social reintegration, a tool developed to measure the degree of adjustment to community living in persons with SCI. Structural equation modeling was used to examine the direct and indirect effects of self-esteem, social barriers, physical function, family support, informational support, perceived stress, emotion-focused coping (EFC), and problem-focused coping (PFC) on social reintegration. RESULTS: The model explained 65% of the variance in social reintegration in persons with SCI. The results indicated that the social integration of persons with SCI was influenced most by EFC. Family support, informational support, perceived stress and social barriers were also significantly related to social reintegration. CONCLUSIONS: These findings suggest implications for developing the interventions at various levels including family and community and specific to individual coping strategies to enhance social reintegration in persons with SCI. PMID- 15823996 TI - Monitoring standing wheelchair use after spinal cord injury: a case report. AB - PURPOSE: An important issue in spinal cord injury (SCI) research is whether standing can yield positive health benefits. However, quantifying dose of standing and establishing subject compliance with a standing protocol is difficult. This case report describes a method to monitor dose of standing outside the laboratory, describes the standing patterns of one subject, and describes this subject's satisfaction with the standing protocol. METHOD: A man with T-10 complete paraplegia agreed to have his commercially available standing wheelchair instrumented with a custom-designed logging device for a 2-year period. The micro-controller-based logger, under custom software control, was mounted to the standing wheelchair. The logger recorded date, duration, angle of standing, and start/stop times. RESULTS: The client exceeded a suggested minimum dosage of standing per month (130.4% of goal), choosing to stand for short bouts (mean = 11.57 min) at an average angle of 61 degrees, on an average 3.86 days per calendar week. He was generally very satisfied with the standing device and provided subjective reports of improved spasticity and bowel motility. CONCLUSION: This case report describes a standing and surveillance system that allow quantification of standing dose. Future controlled studies are needed to evaluate whether standing can be beneficially affect secondary complications after SCI. PMID- 15823997 TI - Survey of caffeine levels in retail beverages in Portugal. AB - The caffeine content of 85 retail beverage samples purchased from local supermarkets between 1995 and 2004 was determined. The potential intake of caffeine through the consumption of these beverages (but excluding coffee) was estimated for students of the University of Coimbra, Portugal. The caffeine content of the beverages ranged from 47.5 to 282.5 mg l(-1) for teas, from 20.1 to 47.2 mg l(-1) for tea extracts samples, and from 80.7 to 168.7 mg l(-1) for cola soft drinks. Caffeine was not completely absent from caffeine-free colas, and energy drinks had a far greater caffeine content than regular drinks, ranging from 21 to 2175 mg l(-1). Soft drinks were consumed by 72% of the individuals, although 14% of the survey participants did not drink any of the different types of the beverages studied. Contrary to expectations for this age group, no consumptions of energy drinks was reported. Daily caffeine intake was estimated to range from 4.7 to 200 mg day(-1), but with only 5% reporting a daily intake around 200 mg caffeine. Cola-type beverages were an important dietary source of caffeine for the population studied. Statistical differences in the caffeine intake between the male and female populations were found, with p = 0.014, being higher for the male population. Of the beverages studied, cola-type drinks showed statistical differences for the male population, p = 0.03, and tea showed statistical differences for female population p = 0.013, respectively. PMID- 15823998 TI - Influence of dough ingredients on 3-chloropropane-1,2-diol (3-MCPD) formation in toast. AB - The influence of different dough ingredients such as fat, salt, sourdough, emulsifiers, and sugar on the formation of 3-chloropropane-1,2-diol (3-MCPD) during toast preparation under domestic conditions was investigated. In comparison with a fat-free recipe, addition of 1% peanut fat considerably increased 3-MCPD formation, but varying the fat (2-5%) or salt (1.6-2.4%) contents within technological acceptable limits did not show any significant differences. A baking agent, which is usually commercially applied by many toast bakers or industrial toast manufacturers, increased 3-MCPD formation in toasted bread slices. Considerable evidence was found that the baking agent's main component sucrose had the major part in increasing 3-MCPD levels. Emulsifiers containing monoacylglycerols moderately increased 3-MCPD levels, but the addition of lecithin did not have any significant influence. 3-MCPD levels showed a good correlation with the lightness (L* value) of the bread slices; their 3-MCPD content increased exponentially towards dark coloured toasts. The relation between 3-MCPD and 2-MCPD was an average of 3:1 in all samples. Dichloropropanols such as, for example, 1,3-dichloropropanol could not be detected. PMID- 15823999 TI - Inter-laboratory validation study of five commercial ELISA test kits for the determination of peanut proteins in biscuits and dark chocolate. AB - The results of an inter-laboratory study with five commercially available peanut ELISA test kits to detect and quantify peanut residues in two food matrices (biscuit and dark chocolate) at four different concentrations (0-10 mg peanut kg( 1) matrix corresponding to about 0-2.5 mg peanut protein kg(-1) matrix) are reported. In general the five ELISA test kits evaluated could detect peanut protein in the two food matrices. In three cases, the study challenged the test kits beyond their intended use for quantification below the manufacturers' defined cut-off limits. Generally, all five ELISA test kits performed well in the concentration range 5-10 mg kg(-1) rather than in the low concentration range (2.0 or 2.5 mg kg(-1)). The variation in the found recoveries of peanut between the different test kits had a spread of 44-191% across all concentrations. The quantification characteristics between test kits differed significantly at the very low mg kg(-1) level. Two test kits performed well even at concentrations below 5 mg kg(-1) with reproducibilities of 27-36% for biscuits and 45-57% for chocolate. PMID- 15824000 TI - Confirmatory method for the analysis of carbadox and olaquindox in porcine feedingstuffs using LC-electrospray MS-MS. AB - A method is described for the quantitative determination of the two feed additives carbadox and olaquindox in porcine feedingstuffs. The use of these agents in feedingstuffs was prohibited in the European Union as a result of concerns about their toxicity. Regulatory laboratories are required to have suitably validated analytical methods to ensure compliance with the ban. The analytes were extracted from finished feedingstuffs into acetonitrile:chloroform (1:1, v/v), and aliquots (1.0 ml) of the extract were dried down under a stream of nitrogen at 65 degrees C. All residues were re-dissolved in HPLC mobile phase containing acetonitrile/water/formic acid. Analysis was based on LC coupled to positive-ion electrospray MS-MS, with daughter ions for carbadox at m/z 231 and 90, and for olaquindox at m/z 212 and 143 being monitored. The method was validated by analysing feed samples fortified with carbadox and olaquindox at 0.5, 2.5 and 5 mg kg(-1) on three separate occasions. Sample preparation was simple, thus allowing the confirmation of these compounds in large numbers of samples. PMID- 15824001 TI - Analysis of nifursol residues in turkey and chicken meat using liquid chromatography-tandem mass spectrometry. AB - Nifursol (3,5-dinitrosalicylic acid (5-nitrofurfurylidene) hydrazide) is mainly used as a feed additive for the prevention of blackhead disease in turkeys. The objective of the present work was to establish information on nifursol residues in turkey and chicken meat. The analytical method was based on conversion of nifursol and its metabolites with an intact 3,5-dinitrosalicylic acid hydrazide (DNSH) side chain to the 2-nitrophenyl analogue of nifursol (NPDNSH) by treatment with dilute hydrochloric acid and 2-nitrobenzaldehyde. Nifuroxazide (salicylic acid (5-nitrofurfurylidene) hydrazide) added as an internal standard was converted to the 2-nitrophenyl analogue NPSH. After the addition of ammonia, proteins were precipitated with acetonitrile. Chromatographic separation was achieved on a C18 column and negative-ion electrospray ionization mass spectrometry was employed using m/z 183 and 226 (daughter ions of the NPDNSH phenolate ion m/z 374) for quantification and m/z 93 (daughter ion of the NPSH phenolate ion m/z 284) as a retention time reference. The decision limit (CCa) and detection capability (CCbeta) of the analytical method were 0.05 and 0.08 microg kg(-1), respectively. In the range 0.5-1 microg kg(-1), the repeatability, within-laboratory reproducibility and trueness were 8, 11 and -1%, respectively. A total of 37 samples of turkey meat and 16 samples of chicken meat were purchased at retail outlets in early spring, summer and winter 2003, and analysed for nifursol residues. No residues were found in the chicken samples, but nine of 18 samples of turkey meat collected in the spring had between 0.05 and 0.6 microg kg(-1) (average 0.25 microg kg(-1)) nifursol residues. PMID- 15824002 TI - Screening for the coccidiostats halofuginone and nicarbazin in egg and chicken muscle: development of an ELISA. AB - Nicarbazin and halofuginone have been widely used as coccidiostats for the prevention and treatment of coccidiosis in poultry. It has been shown that accidental cross-contamination of feed can lead to residues of these compounds in eggs and/or muscle. This paper describes a direct competitive assay for detecting halofuginone and nicarbazin, developed as qualitative screening assay. In an optimized competitive ELISA, antibodies showed 50% binding inhibition at approximately 0.08 ng ml(-1) for halofuginone and 2.5 ng ml(-1) for dinitrocarbanilide (marker residue for nicarbazin). Extraction from the matrix was carried out with acetonitrile followed by a wash with hexane. The assay's detection capability (CCbeta) for halofuginone was < 0.5 microg kg(-1) in egg and < 1 microg kg(-1) in muscle. For dinitrocarbanilide, the CCbeta was estimated at < 3 microg kg(-1) in egg and < 10 microg kg(-1) in chicken muscle. PMID- 15824003 TI - Changes in the chemical form of selenium observed during the manufacture of a selenium-enriched sourdough bread for use in a human nutrition study. AB - High-performance liquid chromatography interfaced with inductively coupled plasma mass spectrometry, and hydride generation-inductively coupled plasma mass spectrometry were used, respectively, to investigate changes in both the chemical form and the concentration of selenium during its bio-incorporation and bio accumulation into rye seedlings. A 60-fold increase in the total level of selenium in the seedlings ('control' biomass = 0.99 mg kg(-1), 'enriched' biomass = 55.27 mg kg(-1)) was accompanied by a change from selenite to several organo selenium forms, with more than 40% being present as selenomethionine. The seedling biomass was dried, ground and used as an ingredient in the production of a fermented sourdough bread (popular in Poland and many Eastern European countries). The selenium in the resulting bread was also characterized in terms of its speciation, as well as its total selenium content ('control' bread = 0.06 mg kg(-1), 'enriched' bread = 3.56 mg kg(-1)). The breads were then fed to 24 volunteers as part of a human intervention study designed to establish the efficacy of this mode of selenium supplementation. The human study data subsequently showed the bread was a good source of dietary selenium. PMID- 15824004 TI - Concentrations of heavy metals in the tissues of red deer (Cervus elaphus) from the region of Warmia and Mazury, Poland. AB - Cadmium, lead, copper and zinc were quantified using a validated ICP/MS method in the tissues and organs of 82 specimens of red deer of different sexes and ages, which had been hunted at the Great Lakes Land region in the north-eastern of Poland in 2000-01. Copper and zinc occurred in liver, kidney and muscle tissue of red deer at concentrations of 6.4-29, 3.3-7.2 and 1.9-6.4, and of 19-43, 17-41 and 19-64 mg kg(-1), respectively, which are considered typical for the species, age and gender compared with values noted for some other wild and domestic ruminant species elsewhere. For cadmium and lead, the concentrations decreased in the order kidney > liver > muscle tissue, with overall means of 2.2, 0.19, and 0.10, 0.31, and 0.26, 0.22 mg kg(-1) wet weight, respectively. The cadmium content of the kidney of a proportion of red deer sampled exceeded the legal tolerance limits set in Poland. The means (but not individual animal data) obtained for cadmium in kidney and liver correlated (0.99) with the age of red deer. Also, the lead content of the muscle meat of red deer for many carcasses exceeded the legal limit, which was probably due to contamination from the fine dust particle remains from the lead bullets used. PMID- 15824006 TI - Ochratoxin A: an improvement clean-up and HPLC method used to investigate wine and grape juice on the Polish market. AB - A routine method appropriate for the determination of ochratoxin A (OTA) in wine, grape juice and grape juice drinks was described, and the occurrence of the mycotoxin was investigated in the most popular red wines, grape juice and grape juice drinks available on the Polish market. After clean-up on immunoaffinity column, samples were analysed by RP-HPLC using a fluorescence detector at 330 and 460 nm. The average OTA recoveries from spiked blank wine samples varied from 60 to 82%, and RSD% ranged from 5 to 14%. The OTA recovery for spiked grape juice and grape juice drinks were 80-100%, but the RSD% was between 7 and 10%. The limit of detection and limit of quantitation for all sample types were 0.5 and 2.0 ng l(-1), respectively. Fifty-three samples of red wine and seven samples of grape juice and grape drinks were assessed by means of this analytical procedure. OTA was detected in most wine samples (92%); its concentrations ranged from 2.2 to 6710 ng l(-1). In all grape juice and drink samples, OTA levels ranged from 1.6 to 64.7 ng l(-1). PMID- 15824005 TI - Fate of mycotoxins in cereals during extrusion cooking: a review. AB - Extrusion cooking is one of the fastest growing food-processing operations in recent years due to several advantages over traditional methods. Apart from its main goal of improving the quality of intermediate and final processed products, it may incidentally also improve safety because of the potential to reduce mycotoxin levels in cereals. This review is focused on extrusion cooking and aims to give a general overview of its impact in reducing mycotoxin levels in cereals. Extrusion cooking generally decreases the mycotoxins levels at rates depending on different factors such as the type of extruder, the type of screw, the die configuration, the initial mycotoxin concentration, the barrel temperature, the screw speed, the moisture content of the raw material and the use of additives. Reductions of 100, 95 and 83% for fumonisins, aflatoxins and zearalenone, respectively, have been reported during extrusion cooking of cereals, while lower reductions were observed for deoxynivalenol, ochratoxin A and moniliformin, where maximum reductions did not exceed 55, 40 and 30%, respectively. PMID- 15824007 TI - Analysis of duplicate 24-hour diet samples for aflatoxin B1, aflatoxin M1 and ochratoxin A. AB - In the spring and autumn of 1994, a total diet study, in which 123 participants collected duplicates of their 24-hour diets, was carried out. The goal of this study was to determine the mass fractions of a number of analytes in these duplicate diets, so as to be able to establish oral daily intake values. After measurements were carried out for pesticides, PCBs, elements, sterols, nitrate and nitrite, and fatty acids, the duplicate diet study was concluded with analyses for aflatoxin M1, aflatoxin B1 and ochratoxin A. For this purpose a method of analysis was developed, that could simultaneously determine these mycotoxins at very low levels. The method involved chloroform extraction, liquid liquid extraction, immunoaffinity cleanup and liquid chromatography. The method was supplemented with a procedure to confirm the identity of chromatographic peaks, assumed to represent aflatoxin M1, aflatoxin B1 and ochratoxin A. The method was in-house validated. Recoveries ranged from 68-74% for aflatoxin M1 (at spiking levels from 30-120 ng/kg, c.v. 7.6%), from 95-97% for aflatoxin B1 (at spiking levels from 50-200 ng/kg, c.v. 2.8%), and from 75-84% for ochratoxin A (at spiking levels from 150-600 ng/kg, c.v. 4.3%). Limits of quantitation (defined as signal/noise = 10) were estimated to be 24, 5 and 16 ng/kg lyophilised material for aflatoxin M1, aflatoxin B1 and ochratoxin A respectively. The newly developed method was used to analyse 123 samples of 24 hour diets. Aflatoxin M1 was detectable in 48% of the samples; the toxin contents remained below the limit of quantitation in all samples. Aflatoxin B1 could be detected in 42% of the samples; in 25% of the samples the levels were above the limit of quantitation. Ochratoxin A could be quantified in all samples. The analytical results were further processed to estimate levels of intake. Intake levels for the aflatoxins were very low, and could not reliably be established. The mean ochratoxin A intake was estimated to be 1.2 ng/kg body weight per day. This is well below the tolerable daily intake established by JECFA at 14 ng/kg body weight per day. The current dietary intake of ochratoxin A in the Netherlands is concluded to pose no appreciable health risk. PMID- 15824008 TI - Evolution of aroma compounds from orange juice stored in polyethylene terephthalate (PET). AB - The evolution of aroma compounds from orange juice made from concentrate and stored in glass, standard monolayer polyethylene terephthalate (PET 1), multilayer PET (PET 2) and plasma-treated PET (internal carbon coating) (PET 3) was investigated. Bottles were stored at room temperature (20 degrees C) under artificial light. Volatile compounds in orange juice samples and corresponding packaging materials were analysed at zero time and after 2, 3 and 5 months of storage. After 5 months of storage, from 0.2 to 0.3% of the initial amounts of limonene and beta-myrcene in the orange juice had been absorbed by the plastic packaging materials. Statistical analyses showed that the evolution of aroma compounds was strongly correlated to the duration of storage, but not to the type of packaging material. Indeed, whatever the stored orange juice samples, the same evolutions were observed, with a decrease in aldehydes and ketones, esters, aliphatic alcohols, sesquiterpene and monoterpene alcohols, and an increase in two aliphatic and monoterpene alcohols (i.e. furfural and 4-vinylguaicol). The results suggest that the losses of aroma compounds from the juice could be attributed to the high acidity of the matrix, implying acid-catalysed reactions. Finally, PET packaging materials and their corresponding oxygen permeabilities showed no correlation with the loss of aroma compounds. PMID- 15824009 TI - Magnetic susceptibility artifact in orbital magnetic resonance imaging. AB - Case report of a 16-year-old boy presenting with total hyphema due to blunt trauma to the left eye. During a subsequent cinematic MRI scan to identify salvageable lateral rectus muscle, the attachment of the left lateral rectus was obscured by a 20-mm diameter signal void. A subsequent CT scan for a suspected metallic foreign body revealed the muscle attachment after all. The theoretical explanations for this finding are discussed. PMID- 15824010 TI - Postoperative adjustable surgery of the superior oblique tendon. AB - PURPOSE: To describe a surgical procedure for loosening or tightening the superior oblique (SO) tendon that enables bedside adjustment following the surgery, with surgical outcome reported. METHODS: A permanent suture separates the two cut ends of the SO tendon, with a sliding noose for adjustment. The noose is accessed by having the patient look up or straight ahead, not down, during adjustment at the bedside. Records of 17 patients who underwent this surgery between June 2000 and January 2003 were reviewed and analyzed for outcome. RESULTS: Seventeen patients, 18 eyes, mean age 43.7 years (range 5.9 to 71 years) had SO surgery with postoperatively adjustable sutures. Twelve eyes of 11 patients had a loosening procedure, and six eyes had a tightening procedure. Seven of the patients had had precious eye muscle surgery, four having had previous surgery on the same SO tendon. All but one patient returned for the follow-up examination, ranging from 1.5 to 7 months postoperatively. Torsional imbalances in 12 patients improved in all but two. In four patients with a preoperative A pattern, the A pattern improved from 16 PD to 0 PD on average. In eight patients with vertical misalignment in straight ahead gaze who had no other cyclovertical muscle surgery simultaneously, the mean reduction was 7 PD. All patients except one had improvement in preoperative symptoms. CONCLUSION: Both loosening and tightening procedures can be performed successfully using a postoperative adjustment technique for the SO tendon. PMID- 15824011 TI - Non-traumatic sixth nerve palsy in a young patient. AB - Non-traumatic sixth nerve palsy in the 20-50 years old group is unusual and a detailed history is often needed to identify serious and potentially reversible causes. We present a case of sixth sense nerve palsy in a young female with eczema for which she was treated with low-dose cyclosporin. We review the indications for and ocular side effects of cyclosporin, a drug with which most ophthalmologists will be unfamiliar. PMID- 15824012 TI - Defining amblyopia: the need for a joint classification. AB - Recent years have shown an impressive rise in high quality research on amblyopia. Unfortunately, the condition is not sufficiently defined, and consequently different studies have been used different definitions of amblyopia. Aspects in need of consideration include the degree of visual acuity reduction, magnitude of interocular difference in acuity, method of visual acuity testing used, the effect of refractive adaption, the presence of amblyogenic factors, the absence of organic cause and the treat ability of the deficit. Vision scientists worldwide are encouraged to jointly decide on what is, and what is not, amblyopia. PMID- 15824013 TI - Maximum angle of ocular duction during visual fixation as a function of age. AB - PURPOSE: To measure and compare the maximum angle of ocular duction in healthy individuals as a function of age. METHODS: A calibrated arc perimeter was modified to display one of six randomly presented targets (high contrast Snellen equivalent letters), in both vertical (supra/infraduction) and horizontal (ab/adduction) gaze to the dominant eye of 204 healthy volunteers with best corrected visual acuity. A bite-bar and headrest were employed to prevent head movement. Using a modified method of limits for discrimination threshold, a maximum mean angle of ocular duction was determined by stepping a target out in 5 degrees steps until an error was reported and thereafter bracketing around the limits of the target identification in 1 degrees steps. A mean threshold value was determined as the angle at which a subject obtained a correct response 75% of the time in two and as many as the trails in each of the four randomly presented directions of gaze (abduction, adduction, supraduction and infraduction). RESULTS: A decrease in mean maximum duction angle was found over all age groups in al four directions (p < 0.001), with a step decline beginning in the sixth decade and almost doubling in the oldest age group tested (80-95 years-olds). The percent change in mean maximum angle of duction due to age from the 14-19 to the 80-95 year-olds was: abduction 21%, adduction 24%, supraduction 35%, infraduction 26%. CONCLUSION: Baseline data are useful to differentiate normal changes occurring with age from early signs of disease. AdditIonally, disease progression and effects of treatment can be monitored. PMID- 15824014 TI - Screening for amblyogenic factors in preschool children with the retinomax hand held refractor: do positive children have amblyopia and is treatment efficacious? AB - PURPOSE: To determine the visual acuity (VA) in a group of preschool children who were true positives for refractive screening (positive group) in order to compare it with the VA of a sample of children without any refractive anomaly (control group); also, to investigate if true-positive treated children can achieve the VA of the control group within what delay. METHODS: Fifty seven children without any refractive anomaly were tested with the VA child chart if the Nidek SCP 670 projector; 47 children presenting with the refractive anomalies were tested with the same chart. A Kruskal Wallis exact test for singly ordered tables were used to compare VA between those groups and a non-parametric Wilcoxon test for paired samples was used to compare pre- and post-treatment VA in 27 children of the positive group. The mean time necessary to achieve the post-treatment VA was calculated. RESULTS: Median VA: 20/20 in the control group, 20/33 in the positive group. The VA difference between the two pre- and post-treatment VA are highly significant. Among the 27 treated children, pre-treatment median VA was 20/50, post-treatment 20/20. The differences between the pre- and post-treatment VA was 9 months. CONCLUSION: The positive group had a significantly lower VA than the control group. In the treated group, treatment initiated at a mean age of 4 years resulted in an increase in VA to that of the control group after an average time of 9 months, encouraging early screening and treatment before VA is measureable in order to shorten this delay. PMID- 15824017 TI - Anatomical description of the human eye. PMID- 15824015 TI - Incomitance in monkeys with strabismus. AB - PURPOSE: Rhesus monkeys reared with restricted visual environment during their first few months of life develop large ocular misalignment (strabismus). The purpose of this study was to describe 'A and V' patterns and DVD in these animals during fixation and eye movements and suggest that this form of rearing produces animals that are suitable model to study the mechanisms that might cause 'A/V' pattern incomitant strabismus and dissociated vertical deviation (DVD) in humans. METHODS: Eye movements were recorded during fixation, smooth-pursuit and saccades using binocular search coils in one monkey with esotropia, three monkeys with exotropia and one normal monkey. RESULTS: 1) Monkeys reared with Alternating Monocular Occlusion or Binocular deprivation (tarsal plates intact) showed both horizontal and vertical misalignment during monocular and binocular viewing. 2) Large 'A' patterns were evident in 2 out of 3 exotropes while a 'V' pattern was observed in the esotrope. 3) Similar 'A/V' patterns were observed with either eye viewing and during fixation or eye movements. 4) The vertical misalignment, which consisted of the non-viewing eye being higher than the fixating eye, appeared to constitute a DVD. CONCLUSION: Visual sensory deprivation methods that induce large strabismus also induce 'A/V' patterns and DVD similar to certain types of human strabismus. The source of pattern strabismus could be central, i.e., altered innervation to extraocular muscles from motor nuclei, or peripheral, i.e., altered location of extraocular muscle pulleys. PMID- 15824018 TI - Chemoprevention: mouse colon and lung tumor bioassay and modulation of DNA methylation as a biomarker. AB - Lung and colon tumors were induced in A/J, C3H, and A/J X C3H (AC3) mice by administering 16 mg/kg vinyl carbamate followed by 6 weekly doses of 12 mg/kg azoxymethane (AOM). Beginning 1 week after carcinogen treatment, the mice received chemopreventive agents, dexamethasone or piroxicam, at 0.1 and 75 mg/kg in the diet, respectively. Both AOM and vinyl carbamate induces lung tumors, but only AOM induced colon tumors. The strain sensitivity for both colon and lung tumors was A/J > AC3 > C3H mice. Dexamethasone and piroxicam reduced the multiplicity of colon and lung tumors in A/J and AC3 mice, demonstrating the advantage of a combined colon and lung bioassay. The ability of budesonide, a drug that prevents mouse lung tumors, to modulate DNA methylation in vinyl carbamate-induced lung tumors was also determined. Budesonide administered for only 7 days prior to sacrifice caused a dose-dependent (0.6 to 2.4 mg/kg diet) reversal in tumors of DNA hypomethylation and hypomethylation of the insulin-like growth factor (IGF)-II gene in the differentially methylated region (DMR) 2 region of exons 4 to 5. Longer treatment with budesonide reversed hypomethylation when administered up to the time of sacrifice. These results indicate that reversal of the hypomethylation of DNA and of specific genes in lung tumors may be applicable as a surrogate end-point biomarker for chemoprevention. PMID- 15824019 TI - Identification of genetic polymorphisms through comparative DNA sequence analysis on the K-ras gene: implications for lung tumor susceptibility. AB - In the present study, the authors performed a comparative sequence analysis of the K-ras gene. By comparing sequences from different mouse inbred strains, the authors have identified new nucleotide polymorphisms in the noncoding regions of mouse K-ras gene. They have also identified noncoding DNA segments evolutionarily conserved among the human, mouse, and rat. Computational analysis for transcription factor binding sites suggests that these polymorphic and conserved DNA sequences harbor potential cis-regulatory elements, which may contribute to the transcriptional regulation of the K-ras gene. Further studies on these potential regulatory sites may help to elucidate the fundamental mechanism underlying allele-specific activation and expression of K-ras gene in hybrid mouse lung tumors, which determines lung tumor susceptibility in mice. PMID- 15824020 TI - Characterization of two protein-binding sites in the second intron of the mouse K ras gene. AB - A tandem repeat region in the second intron of the K-ras gene has been reported to be a possible regulatory site for transcription. In this study, a second protein-binding site was identified and characterized. It lies downstream (nucleotides 463 to 509) of the tandem repeat region. A T--> C base variation at nucleotide 494 was found in all K(S) strains (which have K-ras alleles identical to those of susceptible A/J strain) and all K(i) strains (which have K-ras alleles identical to those of the intermediate CBA/J strain). DNase I footprint analysis indicated a protein binding site within the downstream repeated region in the second intron of the K-ras gene. Gel mobility-shift studies showed differential protein-binding patterns between the K(r) strains (which have K-ras alleles identical to those of the resistant C3H/HeJ strain) and the K(s) or K(i) strains. Southwestern blot analysis of DNA-protein complexes indicated that the 2 repeated regions might bind the same regulatory complex. PMID- 15824021 TI - Familial risk for lung cancer by histology and age of onset: evidence for recessive inheritance. AB - The authors used the Swedish Family-Cancer Database to search for evidence for a genetic predisposition in lung cancer. Familial risks in offspring were increased for all lung cancer to 1.77 when a parent was affected with any lung cancers; the comparable risk among siblings was 2.15. At young age, risks between siblings were higher than those between offspring and parents for all histological types of lung cancer. The present data suggest that 1.7% of lung cancers up to age 68 years are heritable and probably due to a high-penetrant recessive gene or genes that predispose to tobacco carcinogens. PMID- 15824022 TI - Pooled analysis on metabolic gene polymorphisms and lung cancer. AB - This article reports an update of pooled analysis on metabolic gene polymorphisms and cancer (the Genetic Susceptibility to Environmental Carcinogens [GSEC] study). The study started in 1997, and has collected data from over 52,000 subjects (half cases, half controls). The distribution of cases shows that the most represented cancer is lung cancer (6465 cases), followed by bladder cancer (3289 cases). The analyses conducted on lung cancer and metabolic gene polymorphisms are summarized and discussed. Pooled analysis allows one to study rare subgroups of subjects, such as lung cancer at young ages, or in nonsmokers, and is a useful approach for generating new working hypothesis and for fostering collaboration. PMID- 15824023 TI - A database of single-nucleotide polymorphisms and a genotyping microarray for genetic epidemiology of lung cancer. AB - The authors set up a database of 105 genes potentially related to lung cancer susceptibility and 464 of their polymorphisms. The database is based on extensive literature searches, and includes genes likely to influence the internal dose of genotoxic compounds that reach the lungs, following exposure to environmental insults, such as tobacco smoke. The authors selected a subset of 250 single nucleotide polymorphisms with appreciable frequency in at least one major ethnic group and/or a clear functional role, which represent the best candidates as lung cancer risk factors. They developed a microarray for genotyping these polymorphisms, based on arrayed primer extension (APEX). PMID- 15824024 TI - The Mouse Tumor Biology Database: integrated access to mouse cancer biology data. AB - Mice have long been used as models for the study of human cancer. The National Cancer Institute has included among its research areas of extraordinary opportunity the development of new mouse genetic models of human cancer and the exploration of cancer imaging as a research tool. Because of the volume and interconnectedness of relevant data, the creation and maintenance of bioinformatics resources of mouse tumor biology is necessary to facilitate current and future cancer research. The Mouse Tumor Biology (MTB) Database provides electronic access to data generated through the study of spontaneous and induced tumors in genetically defined mice (inbred, hybrid, spontaneous and induced mutant, and genetically engineered strains of mice). PMID- 15824025 TI - K-ras mutations in lung tumors from p53 mutant mice exposed to cigarette smoke. AB - In this study, we used p53 transgenic mice to investigate whether mice carrying this germline mutation would be susceptible to tobacco smoke-induced lung tumorigenesis. We subjected male transgenic mice and their wild-type littermates to whole-body exposure to environmental cigarette smoke (ECS) for up to 9.5 months. K-ras gene expression was significantly increased, 28 days after ECS exposure, in the apparently healthy lung of p53 mutant mice. An increase of lung tumor incidence and multiplicity was observed in p53 transgenic mice after exposure to ECS for either 5 months, followed by recovery in air for 4.5 months, or 9.5 continuative months of exposure. Conversely, no tumorigenic effect was observed in their wild-type littermates. Sequence analysis of the K-ras gene indicated that mutations had occurred at codon 12, 13 or codon 61 in tumors both from the air control group and tobacco smoke treatment groups. K-ras mutations were found in 100 %, 100 % and 77 % of tumors from animals exposed to air, ECS for 5 months, followed by recovery in air for 4.5 months, and ECS for 9.5 continuative months, respectively. The K-ras mutations were seemingly not related to the p53 genotype of the animals or to ECS exposure. The mutation spectrum was similar in tumors from the different groups. An apparently higher incidence of K ras codon 12 mutations in the 9.5 months ECS group was not statistically significant. These findings provide evidence that mice carrying a mutant p53 transgene appear to be more sensitive to ECS-induced lung tumors than the corresponding wild-type littermates. K-ras mutations seem to be independent of the p53 status but the early overexpression of this oncogene is related to the p53 status in ECS-exposed mice. These results suggest that tobacco smoke enhances lung tumorigenesis primarily through promoting spontaneously occurring K-ras mutations. PMID- 15824026 TI - Early precursors of low attention and hyperactivity in a preterm sample at age four. AB - The increased numbers of low birth weight (LBW) survivors has raised questions about the direct association between LBW and later diagnosis of Attention Deficit Hyperactivity Disorder (ADHD) in early childhood. A longitudinal data set was used to determine the relationship among perinatal morbidity and medical and neurological status during the toddler period (18 and 30 months) with lower attention and higher activity, cardinal features of ADHD at age 4. The sample of 39 full term and 149 preterm infants were recruited at birth. Infants were assigned to 1 of 5 groups based on perinatal morbidity. Medical and neurological status were classified as normal, suspect, or abnormal at 18 and 30 months. At age 4, five measures of attention and activity were gathered from parents and independent examiners. Multivariate analysis of variance (MANOVA) showed significant effects of perinatal morbidity, birth weight, gestational age, gender, socioeconomic status at infancy, and toddler medical and neurological status with lower attention and higher activity at age 4. Prematurity, perinatal illness, and later medical status are early markers for preschool behaviors associated with clinical diagnosis of ADHD. PMID- 15824027 TI - Occupational risky business: injury prevention behaviors of farm women and children. AB - On farms in the United States, there are approximately 100 fatal and 32,800 nonfatal injuries annually in children 19 years and younger (United States Department of Labor, 1999). Up to 40% of nonfatally injured children are left with permanent disabilities. The impact of injury and death on children associated with farming in the United States is substantial. Research suggests that modeling of health behaviors may be an effective technique for the socialization of children's health behaviors. The purpose of this study is twofold. First, the study describes the participation and use of protective farm equipment or practices by the caregiver and the child. The second purpose is to compare the practices of the caregivers and the children in relation to the use of protective farm equipment and practices. Descriptive and categorical data analysis methods were used to examine the associations of 177 pairs of caregivers and children and their use of protective equipment and preventative behaviors. Greater than 50% of the children under the age of 7 had handled or touched livestock, as well as rode as passengers on tractors. Many children in all age categories had ridden in the back of pick-up trucks. Modeling of the prevention behavior by the caregivers did parallel the use of the equipment by the children. However, the opposite also was true; if the caregiver did not use the equipment, neither did the child. PMID- 15824028 TI - An exploratory study of future plans and extracurricular activities of transition age youth and young adults. AB - A descriptive profile of the health related concerns, school-related and extracurricular activities, employment-related activities, social relationships and future plans of 25 transition-aged youth and young adults (ages 14 to 21 years) were conducted. The findings of this exploratory study provide insight on the impact their chronic condition had upon all aspects of their lives. Sixty percent of respondents indicated they had missed school due to their condition and a third of the respondents had not completed high school. Health care professionals usually were not identified as participatory in youth transition planning. The majority of these transition-aged youth and young adults had positive feelings towards their school experience, although most of the respondents were not involved in school projects or clubs, which suggests their participation in school-related extra-curricular activities was limited. Nearly all of the respondents had some form of employment experience. Most of their work experiences were nonpaying jobs such as serving as a volunteer and participating in school-based employment training. Nearly all of the respondents expressed desires for sustainable employment and fiscal and social independence. Most of the respondents reported having social relationships with just less than half reporting seeing friends outside of school. PMID- 15824029 TI - Childhood suicide: a major issue in pediatric health care. AB - Although prevention of youth suicide is a major aim in most countries, it often is not taken into account that younger children also are capable of killing themselves. Evidence suggests that the suicide rate among children has increased dramatically and that risk factors for suicide must be evaluated when dealing with children in primary and secondary care. Adverse life events in combination with other factors, such as depression, may lead to suicide. This article aims at stimulating further discussion among those involved in pediatric health care. Epidemiological data, an overview of risk factors, and the authors' reflections on the topic are provided. PMID- 15824031 TI - Amplification of antibody responses to antigen using small multilamellar vesicles for delivery system. AB - New vaccines based on subunits, synthetic peptides, or DNA need innovative adjuvants or antigen delivery systems: Spherulites are multilamellar vesicles made of incompatible lipid bilayers without any aqueous core. In this study, we evaluated their ability to induce immune responses against human serum albumin (HSA). Mice were immunized by the intraperitoneal/intravenous route or subcutaneously with HSA without any adjuvant or in its encapsulated form. We showed that Spherulites strongly enhanced the seric antibody responses and led to a mixed isotypic distribution characterized by an IgG(2a) potentiation. This demonstrated that Spherulites can improve the presentation of weak antigens to the immune system. PMID- 15824032 TI - Sorbitan monopalmitate-based proniosomes for transdermal delivery of chlorpheniramine maleate. AB - A proniosomal gel for transdermal drug delivery of chlorpheniramine maleate (CPM) was developed based on Span 40 and extensively characterized in vitro. The system was evaluated for the effect of composition of formulation, type of surfactants and alcohols on the drug loading, rate of hydration, vesicle size, polydispersity, entrapment efficiency, and drug release across cellulose nitrate dialysis membrane. The stability studies were performed at 4 degrees C and at room temperature. The results showed that lecithin produced more stable and larger vesicles with higher loading efficiency but lower dissolution efficiency than cholesterol (chol) and dicethyl phosphate (DCP). The type of alcohol had no significant effect on the stability of vesicles, but ethanol produced larger vesicles (approximately equal to 44 microm) and entrapped a greater amount of drug. Drug release from vesicles of lecithin followed a first-order kinetics whereas those with DCP or without lecithin fit better with a Higuchi model. The proniosomes that contained Span 40/lecithin/chol prepared by ethanol showed optimum stability, loading efficiency, and particle size and release kinetic suitable for transdermal delivery of CPM. PMID- 15824033 TI - Liposomes as alternative vehicles for sun filter formulations. AB - The aim of our study was to determine the influence of several types of liposomes with a different lipid composition on the percutaneous absorption of one conventional sun filter with a lipophilic character (ethyl hexyl methoxycinnamate) using both in vitro and in vivo methodologies. Three different liposomes were prepared with unsaturated and saturated phosphatidylcholine (PC, HPC), and with a wool lipid mixture (IWL) with a composition similar to that of the stratum corneum lipids. Results showed that the liquid crystalline state associated with PC liposomes plays a key role in enhancing skin penetration. when liposomes with a composition and structural organization similar to that of the stratum corneum lipids (HPC and IWL) are used, the skin penetration is retarded, suggesting a certain reinforcement of the stratum corneum barrier. These two types of liposomes could be regarded as alternatives to conventional oil/water emulsions in the formulations of lipidic sun filters. Finally, an acceptable correlation was obtained using both in vitro and in vivo methodologies to evaluate the corresponding skin absorption profile. PMID- 15824034 TI - Salicylic acid and PEG-contained polyanhydrides: synthesis, characterization, and in vitro salicylic acid release. AB - Poly [bi(o-carboxyphenyl)adipate-polyethylene glycol] anhydrides--P(BOCA-PEG)- polymeric drugs were synthesized and characterized by Fourier transformed infrared spectroscopy, NMR, DSC, gel permeation chromatography, etc. Salicylic acid loading efficiency of these polymers ranged from 43.5% to 71.3%, which was much higher than that of other polymeric drugs with salicylic acid. The in vitro release of salicylic acid from the polymers was carried out in buffer conditions with different pH values and and rat gastrointestinal contents. The results showed the release rate of salicylic acid increased with the increase of PEG content in the polymers and the increase of pH value of degradation buffer solution. The rat cecal contents also greatly promoted the release of salicylic acid. In 0.1M phosphate buffer solution at pH8.0, 37 degrees C containing 5% rat cecal contents, P(BOCA-PEG200)(80:20) had 15% salicylic acid released in 21 hr, indicating its potential use in colon-specific salicylic acid delivery. PMID- 15824035 TI - Systematic delivery of chloroquine and promethazine using pH-sensitive polymers. AB - Two pH sensitive polymers (Eudragit L30 D55 and L100) were used as coating materials, respectively, for promethazine hydrochloride and chloroquine phosphate granules formulated with sodium carboxylmethylcellulose and Carbopol 940, respectively, in the ratios 1:1, 1:2, 1:3, and 1:4 (drug:polymer). The granules were characterized. Release studies for the uncoated and coated particles were studied in simulated gastric fluid and simulated intestinal fluid. Result obtained showed that 1:1 and 1:2 ratios of both coated and uncoated granules of the two drugs had short release times and could be recommended for rapid action, whereas 1:4 ratio with low release time could be used for sustained effect. The two granules could be used at varying ratios to obtain desired release characteristics, such that therapeutic concentrations of the two drugs could be achieved. PMID- 15824036 TI - Manufacturing techniques of biodegradable implants intended for intraocular application. AB - Polylactic acid and polylactic-co-glycolic acid are biocompatible and biodegradable polymers with wide utility for the design of controlled release systems for drugs. Regarding intraocular application, polymeric sustained-drug release systems are being studied to treat vitreoretinal diseases. Our work aimed to compare the influence of two implant manufacturing techniques, compression and hot molding, on the in vitro degradation of the polymeric matrices and on the release of dexamethasone acetate. The results showed that the manufacturing technique highly influences degradation and drug release processes. The compressed systems degraded faster and allowed one faster release of the drug. PMID- 15824037 TI - Differential arterial/venous expression of NG2 proteoglycan in perivascular cells along microvessels: identifying a venule-specific phenotype. AB - OBJECTIVE: Similar to other vascular pericyte markers, including smooth muscle (SM) alpha-actin, desmin, and PDGF-beta-receptor, NG2 proteoglycan is not pericyte specific. Therefore, the use of NG2 as a pericyte marker, especially in cell lineage studies, in comparison to other nonspecific pericyte markers requires an understanding of how its expression varies spatially within a microvascular network. The objective of this study was to characterize NG2 expression along vessels within rat microvascular networks and compare this to SM alpha-actin expression. METHODS: Mesenteric tissue, subcutaneous tissue, spinotrapezius muscle, and gracilis muscle were harvested from 250-g, female, Sprague-Dawley rats and stained for NG2 and SM alpha-actin. The distribution of NG2 expression was evaluated in mesenteric networks (n = 28) with complementary observations in subcutaneous tissue and skeletal muscle. RESULTS: Perivascular cells, including mature smooth muscle cells (SMCs), immature SMCs, and pericytes, expressed NG2. Most importantly, NG2 expression was primarily confined to perivascular cells along arterioles and capillaries, and continuous expression was not observed along venules beyond the immediate postcapillary vessels. The differential expression of NG2 along the arteriolar side of microvascular networks was also observed in rat subcutaneous and skeletal muscle. CONCLUSIONS: The results indicate that NG2 is expressed by all perivascular cells along arterioles, and its absence denotes a venule-specific phenotype. These results identify for the first time a marker that differentiates venous smooth muscle and pericytes from other capillary- and arteriole-associated perivascular cells. PMID- 15824038 TI - Neurovascular alignment in adult mouse skeletal muscles. AB - OBJECTIVE: Muscle blood flow increases with motor unit recruitment. The physical relationships between somatic motor nerves, which control muscle fiber contraction, and arterioles, which control microvascular perfusion, are unexplored. The authors tested the hypothesis that motor axons align with arterioles in adult skeletal muscle. METHODS: Transgenic mice (C57BL/6 background, n = 5; 10 months of age) expressing yellow fluorescent protein in all motor nerves underwent vascular casting (Microfil). Excised epitrochlearis, gracilis, gluteus maximus, and spinotrapezius muscles were imaged at 380x and 760x and a computer-integrated tracing system (Neurolucida) was used to acquire 3 dimensional digital renderings of entire arteriolar and neural networks within each muscle. RESULTS: Arteriolar networks were typically approximately 3-fold longer than neural networks. Nerves coursed with arterioles until terminating at motor endplates. Across muscles, proximity analyses revealed that approximately 75% of total nerve length (9.8-48.8 mm) lay within 200 microm of the nearest arteriole (diameters of 15-60 microm). CONCLUSIONS: Somatic motor nerves and arterioles align closely within adult mammalian skeletal muscle. Understanding the signals governing neurovascular alignment may hold important clues for the advancement of tissue engineering and regeneration. PMID- 15824040 TI - Oxygen partial pressure in outer layers of skin: simulation using three dimensional multilayered models. AB - OBJECTIVE: Papillary loops in dermal papillae are believed to facilitate oxygen delivery to the germinal layer in the epidermis. The study aims to simulate the distribution of the oxygen partial pressure, pO2, in superficial layers of skin and to quantify oxygen supply from different sources. METHODS: A theoretical model that considered heterogeneity in tissue properties was developed based on the anatomical arrangement of the upper skin. Distribution of pO2 in tissues and in blood was solved numerically. Effects of the blood flow rate and the skin surface condition were investigated. RESULTS: Under normal blood flow rate and skin surface conditions, approximately one-quarter of the oxygen consumed in the upper skin came from papillary loops. Subpapillary plexus was the main supplier, which accounted for more than 60% of the total oxygen supply. Oxygen diffusion into the skin from the air was restricted to superficial regions of the germinal layer and accounted for approximately 10% of the total oxygen uptake. CONCLUSIONS: Papillary loops in dermal papillae facilitate oxygen delivery to the germinal layer from the circulation. The flow rate in papillary loops affects pO2 distribution in dermal papillae. In the epidermis, however, pO2 is strongly affected by skin surface conditions. PMID- 15824039 TI - Membrane hyperpolarization is not required for sustained muscarinic agonist induced increases in intracellular Ca2+ in arteriolar endothelial cells. AB - OBJECTIVE: Hyperpolarization modulates Ca2+ influx during agonist stimulation in many endothelial cells, but the effects of hyperpolarization on Ca2+ influx in freshly isolated arteriolar endothelial cells are unknown. Therefore, the purpose of the present study was to characterize agonist-induced Ca2+ transients in freshly isolated arteriolar endothelial cells and to test the hypothesis that membrane hyperpolarization augments agonist-induced Ca2+ influx into these cells. METHODS: Arterioles were removed from hamster cremaster muscles and arteriolar endothelial cells were enzymatically isolated. Endothelial cells were loaded with Fura 2-AM and the Fura 2 ratio measured photometrically as an index of intracellular Ca2+. The cells were then stimulated with the muscarinic, cholinergic agonist, methacholine, and the resulting Ca2+ transients were measured. RESULTS: Methacholine (1 microM) increased the endothelial cell Fura 2 ratio from a baseline of 0.81 +/- 0.02 to an initial peak of 1.17 +/- 0.05 (n = 17) followed by a sustained plateau of 1.12 +/- 0.07. The plateau phase of the Ca2+ transient was inhibited by removal of extracellular Ca2+ (n = 12, p < .05), or the nonselective cation channel blockers Gd3+ (30 microM; n = 7, p < .05) or La3+ (50 microM; n = 7, p < .05) without significant effect on the baseline or peak (p > .05). The initial peak of methacholine-induced Ca2+ transients was inhibited by the IP3-receptor antagonist xestospongin D (10 microM, n = 5, p < .05). The methacholine-induced Ca2+ transients were accompanied by endothelial cell hyperpolarization of approximately 14-18 mV, as assessed by experiments using the potentiometric dye, di-8-ANEPPS as well as by patch-clamp experiments. However, inhibition of hyperpolarization by blockade of Ca2+-activated K+ channels with charybdotoxin (100 nM) and apamin (100 nM) (n = 5), or exposure of endothelial cells to 80 or 145 mM KCl (both n = 7) had no effect on the plateau phase of methacholine-induced Ca2+ transients (p > .05). CONCLUSIONS: Freshly isolated arteriolar endothelial cells display agonist-induced Ca2+ transients. For the muscarinic agonist, methacholine, these Ca2+ transients result from release of Ca2+ from intracellular stores through IP3 receptors, followed by sustained influx of extracellular Ca2+. While these changes in intracellular Ca2+ are associated with endothelial cell hyperpolarization, the methacholine-induced, sustained increase in intracellular Ca2+ appears to be independent from this change in membrane potential. These data suggest that arteriolar endothelial cells may possess a novel Ca2+ influx pathway, or that the relationship between intracellular Ca2+ and Ca2+ influx is more complex than that observed in other endothelial cells. PMID- 15824041 TI - A role for hyaluronan in the preservation of interstitial structure. AB - Hydraulic resistance of interstitium is of major importance in body fluid distribution. In the synovial lining it is vital for the retention of intra articular fluid, and is attributed chiefly to the network of interstitial biopolymers occupying intercellular gaps in the tissue. Selective removal of synovial hyaluronan (HA) by protease-free hyaluronate lyase results in an almost 10x increase in synovial hydraulic permeability from 0.48 +/- 0.24 microL min(-1) cm H2O (control) to 4.56 +/- 0.40 microL min(-1) cm H2O (mean +/- SD, n = 6 rabbits, p < .001, t test) leading to the hypothesis that hyaluronan plays a major role in the organization of interstitial matrix structure. To test whether removal of hyaluronan causes significant changes in synovial ultrastructure, morphometry of hyaluronidase-treated synovium was carried out. Following hyaluronidase, the thickness of the synovial lining was reduced from 13.0 +/- 1.6 microm (control) to 10.6 +/- 1.6 microm (mean +/- SD throughout, n = 50 measurements per rabbit, 6 rabbits. p < .001, t test). This was accompanied by a significant reduction of synovial interstitial volume fraction from 76.2 +/- 20.6% (control) to 67.04 +/- 24.94% (p < .001, t test), and an increase in collagen bundle volume as a fraction of interstitial volume from 40.75 +/- 4.97% (control) tissue to 48.77 +/- 11.72% (p < .0001, t test). The findings indicate that the removal of hyaluronan chains leads to morphological disruption. Thus, hyaluronan chains play a major role in the organization of synovial structure. The observed morphological changes are insufficiently large to explain fully the great rise in hydraulic permeability observed on HA removal. The latter is likely to be due to disruption of tertiary architecture at the molecular organization level. PMID- 15824042 TI - Focal adhesion kinase in neutrophil-induced microvascular hyperpermeability. AB - OBJECTIVE: Recent experimental evidence indicates an essential role of focal adhesion kinase (FAK) in mediating endothelial adhesion, contraction, and migration under physical stress and chemical stimulation. However, the functional impact of FAK on microvascular barrier property during inflammation has not been revealed. The aim of this study was to explore the potential contribution of FAK to neutrophil-dependent microvascular hyperpermeability. METHODS: The apparent permeability coefficient of albumin was measured in intact, isolated porcine coronary venules during stimulation by C5a-activated neutrophils. In parallel, the transendothelial flux of albumin was quantified in cultured venular endothelial cell monolayers exposed to C5a-activated neutrophils. Western blotting and immunocytochemistry were performed to assess FAK tyrosine phosphorylation and distribution in endothelial cells, respectively. To specify the signaling effect of FAK on neutrophil-elicited endothelial hyperpermeability, FAK-related nonkinase (FRNK) was expressed, purified, and directly transfected into the endothelium of venules, and the permeability response to neutrophils was measured during inhibition of FAK. RESULTS: C5a-activated neutrophils induced a time- and concentration-dependent increase in venular permeability. Transfection of venules with FRNK did not alter the basal barrier function but greatly attenuated neutrophil-induced hyperpermeability in a dose-related manner. A similar permeability response to neutrophils was observed in venular endothelial cell monolayers, which was diminished after FRNK transfection. In addition, Western blot analysis showed that activated neutrophils caused a concentration dependent increase in FAK tyrosine phosphorylation with a time course correlating with that of venular hyperpermeability. Transfection of FRNK blocked neutrophil evoked FAK tyrosine phosphorylation. Furthermore, immunofluorescence microscopy revealed a significant morphological change of FAK from a punctuated, dot-like pattern under normal conditions to an elongated, dash-like staining that aligned with the longitudinal axis of cells upon neutrophil stimulation. CONCLUSION: The results suggest that focal adhesion kinase significantly contributes to the endothelial hyperpermeability response to neutrophil activation. Phosphorylation of FAK may play an important signaling role in the regulation of microvascular barrier function during inflammation. PMID- 15824044 TI - The adoption of tobacco dependence treatment by rehabilitation clinicians. AB - PURPOSE: Individuals with disabilities tend to smoke at rates that surpass those of the general population. The Pubic Health Service Guideline on the treatment of tobacco dependence suggests that all smokers be screened at every health care visit and counselled regarding how best to quit smoking. We review the literature on the adoption of tobacco dependence treatment by rehabilitation clinicians working with disabled individuals. Despite the deleterious health effects of smoking on individuals with disabilities, the limited data suggests that rehabilitation clinicians rarely encourage their clients who smoke to quit. METHOD: Studies were collected using various computerized databases from 1980 to the present. Because of the paucity of literature on tobacco dependence treatment utilization among rehabilitation clinicians, we also examine research on the use of tobacco dependence treatment by health care providers in the general population and in substance abuse treatment settings. RESULTS: Despite the efficacy of tobacco dependence treatment in smokers with disabilities, tobacco dependence treatment appears to be underutilized by rehabilitation clinicians. CONCLUSIONS: Interventions that have successfully increased adoption by the two other clinician groups should be utilized to increase tobacco dependence treatment provision by rehabilitation clinicians. Additional research is warranted to determine how to overcome obstacles to adoption. PMID- 15824045 TI - Measurement properties of the Activities-specific Balance Confidence Scale among individuals with stroke. AB - PURPOSE: To examine the reliability and validity of the Activities-specific Balance Confidence (ABC) scale among individuals with stroke. METHOD: Descriptive measurement study using a 4-week test-retest design. Data were collected at a tertiary rehabilitation centre form a community-dwelling sample of ambulatory older adults who sustained one stroke at least 1 year prior to the study. During the first measurement session, the total sample (n = 77) completed the ABC as well as the Berg Balance Scale (BBS) and gait speed. The reliability sample (n = 24), a subset of the larger data set, completed the ABC again 4 weeks later. RESULTS: Internal consistency of the ABC was 0.94 and test-retest reliability was ICC = 0.85 (95% CI, 0.68, 0.93). A moderate significant positive, linear correlation with both the BBS (rho = 0.36, P < 0.001) and gait speed (rho = 0.48, P < 0.001) was observed. A factor analysis using principal component analysis indicated that the ABC measures two components of balance self-efficacy (component 1 = perceived low-risk activities; component 2 = perceived high-risk activities). CONCLUSION: The ABC has acceptable measurement properties as demonstrated in this sample of individuals with stroke. This study provides further support for the use of the scale. PMID- 15824046 TI - Foot pain and disability in individuals with Ehlers-Danlos syndrome (EDS): impact on daily life activities. AB - PURPOSE: To investigate the amount and severity of podiatric problems in individuals with EDS as well as the consequences on daily life activities, and to compare these results with the results from a comparison group of the general population. METHOD: The Manchester Foot Pain and Disability Schedule (MFPDS) and a demographic and foot-specific form were mailed. RESULTS: One hundred and thirty six individuals with EDS and 292 from the comparison group responded. The median MFPDS total score in the EDS group was 33 (range 17-49) (mean 32, SD 8) and in the comparison group 17 (range 17-41) (mean 19, SD 4) (P < 0.001). The items most frequently marked with 3 points in the EDS group were: 'I avoid standing for a long time', 'I catch the bus or use the care more often', and 'I get irritable when my feet hurt' (56, 46 and 43%, respectively). When comparing the NRS score (i.e. Numeric Rating Scale) regarding pain intensity between the EDS group (median 5) and the comparison group (median 0), a statistically significant difference was found (P < 0.001). Other foot problems than pain were reported by 73% (n = 100) of the EDS group and by 21% (n = 60) of the comparison group (P < 0.001). The presence of flat feet in the EDS group and the comparison group was reported by 55% (n = 75) and 8% (n = 23) respectively. CONCLUSION: Individuals' with EDS endure difficulties with their mobility due to their foot problems and related disability. It is therefore of utmost importance to find ways to assess the actual foot status for this group and to support the individuals with solutions to their problems with ambulation. PMID- 15824047 TI - Motor and functional recovery in patients with post-stroke depression. AB - PURPOSE: Depression is very common following stroke. Correlation between post stroke depression (PSD) and functional outcome has been shown, but differential impact both on functional and motor recovery has not been deeply investigated. This study evaluates the influence of PSD on motor and functional outcome. METHOD: One hundred and seventeen acute stroke patients were selected in an intensive rehabilitation department, and divided into two groups according to the presence of PSD (PSD+ and PSD-). Screening measures were DSM-IV criteria, the Geriatric Depression Scale and the Cornell Scale. Outcomes were evaluated on the basis of the Barthel Index (BI) and the Fugl-Meyer Assessment Scale (FMA). Measurements were performed at admission to the department T1), discharge (T2) and follow up (T3) in a whole period of 3 months from stroke. RESULTS: Both groups showed a significant improvement in all outcome measures. Improvement differences were not significant on FMA scores in either group at each assessment; the PSD group had a significant higher improvement on BI score at follow-up. According to the logistic model, from T1 to T2 and from T1 to T3, only motor recovery shows a significant relation with functional recovery; from T2 to T3 PSD is the only significant factor related to functional recovery. CONCLUSIONS: PSD is not an influencing factor for motor recovery. Results show a negative impact of PSD on the functional recovery process after discharge and not during hospitalisation. Discharge appears to be critical step for management of PSD. PMID- 15824048 TI - The lowest effective dose of botulinum A toxin in adult patients with upper limb spasticity. AB - OBJECTIVE: To define the lowest effective dose of botulinum toxin type A (Dysport) and safety in the treatment of adult patients with upper limb spasticity. DESIGN: This was a prospective, randomized, double-blind, dose ranging study. Patients received either a placebo or one of three does of Dysport (350, 500 100) U) into five muscles of affected arm by anatomical and electromyography guidance. Efficacy was assessed periodically throughout the 6 month study period by the Modified Ashworth Scale (MAS), the Action Research Arm Test (ARA), the Barthel Index (BI) and the Visual Analogue Pain Scale (VAS). RESULTS: fifty patients were recruited. The four study groups were comparable at baseline with respect to their demographical characteristics and severity of spasticity. All doses of Dysport studied showed a significant reduction from baseline of muscle tone and pain compared to placebo. However, the effect of functional disability was best at a dose of 500 U and the peak improvement was at week 8 after injection. A dose of 1000 U Dysport produced such an excess degree of muscle weakening that the number of randomized patients was reduced to five. BI and ARA of all patients were decrease after injection. No other adverse event was considered related to the study medication. CONCLUSION: This study suggest that treatment with Dysport reduces muscle tone in adult patients with upper limb spasticity. The optimal dose for treatment of patients with residual voluntary movement in the upper limb appears to be 500 U. PMID- 15824049 TI - Shake-It-Up: health promotion and capacity building for people with spinal cord injuries and related neurological disabilities. AB - PURPOSE: This is a description of a model demonstration project called Project Shake-It-Up. This project promotes the health and empowerment of people with spinal cord injuries and related neurological disabilities. It also builds the capacity of community organizations that provide services to this population. METHODS: Participants are involved in interactive seminars concerning health and advocacy on topics like nutrition and being assertive with healthcare providers. They take part in physical and recreational activities such as seat kayaking, kite-flying and hand cycling. Case coordinators support their project participation and personal goal attainment. RESULTS: Community organizations and university researchers developed a successful partnership. Individual members of the first cohort supported one another and benefited physically and psychologically. CONCLUSIONS: Project-Shake-It-Up provides positive initial signs of the value of combining the resources of universities and community agencies. Working together, these organizations can develop distinctive, multi-faceted programmes to support the health and empowerment of people with spinal cord injuries and other related neurological disabilities. PMID- 15824050 TI - Issues for selection of outcome measures in stroke rehabilitation: ICF Body Functions. AB - PURPOSE: To evaluate the psychometric and administrative properties of outcome measures assigned to the ICF Body Functions category, and commonly used in stroke rehabilitation research. METHOD: Critical review and synthesis of measurement properties for five commonly reported instruments in the stroke rehabilitation literature. Each instrument was rated using the eight evaluation criteria proposed by the UK Health Technology Assessment (HTA) programme. The instruments were also assessed for the rigour with which their reliability, validity and responsiveness were reported in the published literature. RESULTS: The reporting of specific measurement qualities for outcome instruments was relatively consistent across measures located within the same general ICF category. Far less information was available on the responsiveness of measures, compared with reliability and validity. The best available instruments were associated with the following body functions: cognitive impairment, depression and motor recovery. CONCLUSIONS: The reader is encouraged to examine carefully the nature and scope of outcome measurement used in reporting the strength of evidence for improved body functions in stroke rehabilitation since there is significant diversity. However there appears to be good consensus about what are the most important indicators of successful rehabilitation outcome in each domain of body function. PMID- 15824053 TI - How to avoid long-term sickness absence: the advice from women with personal experience. AB - OBJECTIVE: The aim of this study was to describe women's perceptions of what can be done to avoid extended sickness absence with following suffering and passivity. METHODS: Qualitative interviews were conducted with 82 women who had been on sickness absence (60 days or more) or were receiving disability pensions. The data were analysed using phenomenological methods. RESULTS: To be able to get back to work was found to be equivalent to breaking away from the prospect of isolation and loneliness. To support this, four parties were identified along with suggestions for their actions: the healthcare professionals, the woman who is on sick leave herself, the employer, and the social insurance official. Most interestingly, the family and close relatives were almost not mentioned at all. The results are connected to a theoretical model of distress in terms of enduring and suffering. CONCLUSIONS: It is necessary to look more carefully at how women on sickness absence use the resources in the world (like their families) to get well. More generally, the task is to understand why society deals insufficiently with women who need time off and cannot keep up with their duties because of illness. PMID- 15824054 TI - A comparative study on attitudes, mental health and job stress amongst GPs participating, or not, in a rural out-of-hours co-operative. AB - OBJECTIVES: To test the study hypothesis that GPs participating in co-operatives will have more positive attitudes towards co-operatives, better mental health and less stress than GPs using traditional out-of-hours arrangements. METHODS: A comparative questionnaire study was conducted amongst GPs, participating, or not, in an out-of-hours, largely rural, co-operative ('NoWDOC') which had been established one year previously. The general attitudes of GPs towards out-of hours work were obtained together with responses to the General Health Questionnaire-12 (mental health) and Stress Arousal Checklist (job stress). RESULTS: Eighty-nine of 120 eligible practitioners responded (74%). The mean GHQ scores for GPs in NoWDOC was 10.2 [standard deviation (SD) 3.9] compared to a score of 11.3 (SD 4.5) for those not participating (t = -1.18; P = 0.24). The overall mean stress score for members of NoWDOC was 3.8 (SD 2.6) compared to 3.4 (SD 2.7) for non-NoWDOC (t = 0.59; P = 0.55). The overall mean arousal score for NoWDOC GPs was 5.2 (SD 2.0) compared to 5.5 (SD 2.9) for non-NoWDOC GPs (t = 0.68; P = 0.50). Multiple regression analyses suggested that the independent variables (partnership arrangements, age, working hours and membership of NoWDOC) did not account for any of the variability in the GHQ score but a significant amount of variability in stress and arousal scores. CONCLUSIONS: The anticipated differences in mental health and job stress among participating GPs were not shown. As the new generation of GPs resemble the NoWDOC participants in their preferences for multi-partner practices with limited out-of-hours care provision, clarification of these findings is important. PMID- 15824055 TI - Does 'peer coaching' increase GP capacity to promote informed decision making about PSA screening? A cluster randomised trial. AB - BACKGROUND: Very little effort has been directed to enable GPs to better informed decisions about PSA screening among their male patients. OBJECTIVES: To evaluate an innovative programme designed to enhance GPs' capacity to promote informed decision making by male patients about PSA screening. METHODS: The study design was a cluster randomised controlled trial set in New South Wales, Australia's most populous state. 277 GPs were recruited through a major pathology laboratory. The interventions were three telephone-administered 'peer coaching' sessions integrated with educational resources for GPs and patients and the main outcome measures were: GP knowledge; perceptions of patient involvement in informed decision making; GPs' own decisional conflict; and perceptions of medicolegal risk. RESULTS: Compared with GPs allocated to the control group, GPs allocated to our intervention gained significantly greater knowledge about PSA screening and related information [Mean 6.1 out of 7; 95% confidence interval (CI) = 5.9-6.3 versus 4.8; 95% CI = 4.6-5.0; P < 0.001]. They were less likely to agree that patients should remain passive when making decisions about PSA screening [Odds ratio (OR) = 0.11; 95% CI = 0.04-0.31; P < 0.001]. They perceived less medicolegal risk when not acceding to an 'uninformed' patient request for a PSA test (OR = 0.31; 95% CI 0.19-0.51). They also demonstrated lower levels of personal decisional conflict about the PSA screening (Mean 25.4; 95% CI 24.5-26.3 versus 27.8; 95% CI 26.6-29.0; P = 0.0002). CONCLUSION: A 'peer coaching' programme, supplemented by education materials, holds promise as a strategy to equip GPs to facilitate informed decision making amongst their patients. PMID- 15824056 TI - A comparison of two methods of collecting economic data in primary care. AB - BACKGROUND: There have been few attempts to assess alternative methods of collecting resource use data for economic evaluations. OBJECTIVE: This study aimed to compare two methods of collecting resource use data in primary care: GPs' case records and a self-complete postal questionnaire. METHODS: 303 primary care attenders were sent a postal survey, incorporating a questionnaire designed to collect service utilisation information for the previous six months. Data were also collected from GP case records. The reporting of GP visits between the two methods, and estimates of costs associated with those visits, were compared. RESULTS: There was good agreement between the number of GP visits recorded on GP case records (mean 3.03) and on the CSRI (mean 2.99) (concordance correlation coefficient = 0.756). In contrast, estimates of average costs of visits from CSRI data were higher and had greater variance compared to case record-based costs (54.63 pound sterling versus 42.37 pound sterling; P = 0.003). This may be explained by differences in average visit length (11.66 versus 9.36 minutes). CONCLUSIONS: This study shows good agreement between GP case records and a self complete questionnaire for the reporting of GP visits. However, differences in costs associated with those visits arose due to differences in the method used for calculating length of visit. PMID- 15824057 TI - Tearing down the Berlin wall: social workers' perspectives on joint working with general practice. AB - BACKGROUND: The arrangements for delivering social work and primary health care to older people in England and Wales are currently subject to rapid re configuration, with the development of integrated primary care and social services trusts. OBJECTIVE: To investigate perceptions of joint working in social services and general practice. METHODS: The study setting was two London boroughs covered by one health authority, one NHS Community Health Services Trust, four Primary Care Groups and two social services departments. All social work team managers in both areas were interviewed together with a purposive sample of social workers with a high number of older clients on their caseloads. A sample of GPs was sought using a sampling frame of practice size in each borough. Structured interviews with open and closed questions were used. Tape-recorded interviews were transcribed and subject to thematic analysis. Analysis of emergent themes was aided by the use of Atlas-ti. RESULTS: Social workers and GPs agree on the need for joint working, but have different understandings of it, each profession wanting the other to change its organizational culture. Co location of social and health care is seen as desirable, but threatening to social work. Concerns about differences in power and hierarchical authority are evident and explicit in social work perspectives. Conflict resolution strategies include risk minimization, adopting pragmatic, case-specific solutions rather than remaining consistent with policy, using nurses as mediators, and resorting to authority. CONCLUSIONS: Although this is a study from urban areas in England, its findings may have wider significance since we have found that resources and professional skills may be more important than organizational arrangements in collaborative working between disciplines. Primary Care Trusts in England and Wales should promote awareness of these different perspectives, perceived risks and conflict minimization strategies in their work on clinical governance and professional development. PMID- 15824058 TI - A dual-light reporter system to determine the efficiency of protein-protein interactions in mammalian cells. AB - Methods for determining protein-protein interactions in mammalian cells typically rely on single reporter functions and are susceptible to variations between samples particularly in regard to levels of transcription, processing and translation. A method has been developed for determining protein-protein interactions in mammalian cells, which bypasses these variables confounding single reporter assays. The approach utilizes two units of gene expression linked to reporter functions that are interposed by a deactivation-activation unit in such a way that the downstream expression unit is switched off. Hence upstream expression occurs regardless of protein-protein interaction, leading to the production of the upstream reporter. In the event of protein-protein interactions, the downstream expression unit is switched on leading to dual reporter read outs. Thus, the ratio of the two reporter activities provides a measure to determine the efficiency of protein-protein interactions. To access the system we screened a mutant of BMPR2 where the interaction between BMPR-II and LIMK is abrogated. BMPR-II is a type II receptor of the TGFbeta superfamily and plays a key role in the pathogenesis of familial pulmonary arterial hypertension. This system has potential for high-throughput screening of libraries (peptide, chemical, cDNA, etc.) to isolate agents that are capable of interfering with highly selective protein-protein interaction. PMID- 15824059 TI - NMR chemical shift and relaxation measurements provide evidence for the coupled folding and binding of the p53 transactivation domain. AB - The interaction between the acidic transactivation domain of the human tumor suppressor protein p53 (p53TAD) and the 70 kDa subunit of human replication protein A (hRPA70) was investigated using heteronuclear magnetic resonance spectroscopy. A 1H-15N heteronuclear single quantum coherence (HSQC) titration experiment was performed on a 15N-labeled fragment of hRPA70, containing the N terminal 168 residues (hRPA701-168) and p53TAD. HRPA701-168 residues important for binding were identified and found to be localized to a prominent basic cleft. This binding site overlapped with a previously identified single-stranded DNA binding site, suggesting that a competitive binding mechanism may regulate the formation of p53TAD-hRPA70 complex. The amide 1H and 15N chemical shifts of an uniformly 15N-labeled sample of p53TAD were also monitored before and after the addition of unlabeled hRPA701-168. In the presence of unlabeled hRPA701-168, resonance lineshapes increased and corresponding intensity reductions were observed for specific p53TAD residues. The largest intensity reductions were observed for p53TAD residues 42-56. Minimal binding was observed between p53TAD and a mutant form of hRPA701-168, where the basic cleft residue R41 was changed to a glutamic acid (R41E), demonstrating that ionic interactions play an important role in specifying the binding interface. The region of p53TAD most affected by binding hRPA701-168 was found to have some residual alpha helical and beta strand structure; however, this structure was not stabilized by binding hRPA701-168. 15N relaxation experiments were performed to monitor changes in backbone dynamics of p53TAD when bound to hRPA701-168. Large changes in both the transverse (R2) and rotating frame (R1) relaxation rates were observed for a subset of the p53TAD residues that had 1H-15N HSQC resonance intensity reductions during the complex formation. The folding of p53TAD upon complex formation is suggested by the pattern of changes observed for both R2 and R1. A model that couples the formation of a weak encounter complex between p53TAD and hRPA701-168 to the folding of p53TAD is discussed in the context of a functional role for the p53-hRPA70 complex in DNA repair. PMID- 15824060 TI - Tissue-dependent isoforms of mammalian Fox-1 homologs are associated with tissue specific splicing activities. AB - An intronic hexanucleotide UGCAUG has been shown to play a critical role in the regulation of tissue-specific alternative splicing of pre-mRNAs in a wide range of tissues. Vertebrate Fox-1 has been shown to bind to this element, in a highly sequence-specific manner, through its RNA recognition motif (RRM). In mammals, there are at least two Fox-1-related genes, ataxin-2 binding protein 1 (A2BP1)/Fox-1 and Fxh/Rbm9, which encode an identical RRM. Here, we demonstrate that both mouse Fxh and A2BP1 transcripts undergo tissue-specific alternative splicing, generating protein isoforms specific to brain and muscle. These tissue specific isoforms are characterized for their abilities to regulate neural cell specific alternative splicing of a cassette exon, N30, in the non-muscle myosin heavy chain II-B pre-mRNA, previously shown to be regulated through an intronic distal downstream enhancer (IDDE). All Fxh and A2BP1 isoforms with the RRM are capable of binding to the IDDE in vitro through the UGCAUG elements. Each isoform, however, shows quantitative differences in splicing activity and nuclear distribution in transfected cells. All Fxh isoforms and a brain isoform of A2BP1 show a predominant nuclear localization. Brain isoforms of both Fxh and A2BP1 promote N30 splicing much more efficiently than do the muscle-specific isoforms. Skeletal muscles express additional isoforms that lack a part of the RRM. These isoforms are incapable of activating neural cell-specific splicing and, moreover, can inhibit UGCAUG-dependent N30 splicing. These findings suggest that tissue specific isoforms of Fxh and A2BP1 play an important role in determining tissue specificity of UGCAUG-mediated alternative splicing. PMID- 15824061 TI - Human Ku70/80 interacts directly with hTR, the RNA component of human telomerase. AB - Maintenance of telomere integrity requires the dynamic interplay between telomerase, telomere-associated proteins and DNA repair proteins. These interactions are vital to suppress DNA damage responses and changes in chromosome dynamics that can result in aneuploidy or other transforming aberrations. The interaction between the DNA repair protein Ku and the RNA component of telomerase (TLC1) in Saccharomyces cerevisiae has been shown to be important for maintaining telomere length. Here, we sought to determine whether this interaction was conserved in higher eukaryotes. Although there is no sequence similarity between TLC1 and the RNA component (hTR) of human telomerase, we show that human Ku70/80 interacts with hTR both in vitro and in a cellular context. Specifically, Ku70/80 interacts with a 47 nt region of the 3' end of hTR, which resembles the stem-loop region of the yeast Ku70/80 binding domain on TLC1. Furthermore, utilizing immunoprecipitation/RT-PCR experiments, we show that Ku interacts with hTR in cell lines deficient in the human telomerase reverse transcriptase protein (hTERT), suggesting that this interaction does not require hTERT. These data suggest that Ku interacts directly with hTR, independent of hTERT, providing evidence for the conservation of the interaction between Ku and telomerase RNA among various species and provide significant insight into how Ku is involved in telomere maintenance in higher eukaryotes. PMID- 15824062 TI - Torque-limited RecA polymerization on dsDNA. AB - The assembly of RecA onto a torsionally constrained double-stranded DNA molecule was followed in real time using magnetic tweezers. Formation of a RecA-DNA filament on the DNA tether was stalled owing to different physical processes depending on the applied stretching force. For forces up to 3.6 pN, the reaction stalled owing to the formation of positive plectonemes in the remaining DNA molecule. Release of these plectonemes by rotation of the magnets led to full coverage of the DNA molecule by RecA. At stretching forces larger than 3.6 pN, the twist induced during filament formation caused the reaction to stall before positive supercoils were generated. We deduce a maximum built-up torsion of 10.1 +/- 0.7 k(b)T. In vivo this built-up torsion may be used to favor regression of a stalled replication fork or to free the chromosomal DNA in E.coli from its condensing proteins. PMID- 15824063 TI - U7 snRNP-specific Lsm11 protein: dual binding contacts with the 100 kDa zinc finger processing factor (ZFP100) and a ZFP100-independent function in histone RNA 3' end processing. AB - The 3' cleavage generating non-polyadenylated animal histone mRNAs depends on the base pairing between U7 snRNA and a conserved histone pre-mRNA downstream element. This interaction is enhanced by a 100 kDa zinc finger protein (ZFP100) that forms a bridge between an RNA hairpin element upstream of the processing site and the U7 small nuclear ribonucleoprotein (snRNP). The N-terminus of Lsm11, a U7-specific Sm-like protein, was shown to be crucial for histone RNA processing and to bind ZFP100. By further analysing these two functions of Lsm11, we find that Lsm11 and ZFP100 can undergo two interactions, i.e. between the Lsm11 N terminus and the zinc finger repeats of ZFP100, and between the N-terminus of ZFP100 and the Sm domain of Lsm11, respectively. Both interactions are not specific for the two proteins in vitro, but the second interaction is sufficient for a specific recognition of the U7 snRNP by ZFP100 in cell extracts. Furthermore, clustered point mutations in three phylogenetically conserved regions of the Lsm11 N-terminus impair or abolish histone RNA processing. As these mutations have no effect on the two interactions with ZFP100, these protein regions must play other roles in histone RNA processing, e.g. by contacting the pre-mRNA or additional processing factors. PMID- 15824064 TI - Alcoholism and personality disorders: an exploratory study. AB - AIMS: To define the most frequent personality disorders related to alcohol dependence. METHODS: Using the International Personality Disorder Examination and the Millon Clinical Multiaxial Inventory-II for personality disorders, 30 consecutively recruited alcohol-dependent patients attending an outpatient clinic were compared with 30 consecutively recruited psychiatric patients with non addictive disorders and 31 subjects from the general population chosen to match the patient samples for age, gender and socio-economic level. RESULTS: Forty percent of the alcohol-dependent patients and 16.6% of the general clinical sample (vs 6.4% of the normative sample) showed at least one personality disorder. Dependent personality disorders were most prevalent (13.3%), followed by paranoid and obsessive-compulsive personality disorders (10% each). PMID- 15824065 TI - Epidemiology of heavy alcohol use in Ukraine: findings from the world mental health survey. AB - AIMS: To describe the epidemiology of heavy alcohol use in Ukraine, using data from the world mental health (WMH) survey in Ukraine. METHODS: The WMH composite international diagnostic interview was administered in 2002 to a national probability sample of Ukrainian adults (n=4725). An algorithm for classifying heavy use in the past year was developed from self-reports about the quantity and frequency of drinking, and its convergent validity was demonstrated. Prevalence rates and socio-demographic risk factors were examined separately for men and women. RESULTS: The 12-month rates of heavy alcohol use were 38.7% in men and 8.5% in women (22.0% overall). Among heavy alcohol users, 92% of men and 52% of women consumed at least 80 g of ethanol in a typical drinking day on a monthly basis in the year before the interview. The most significant risk factors in men and women were age (26-54 years for men; 18-25 years for women), living in the Southeast region, being in the labour force whether employed or unemployed, and for men, low education and being the father of a young child. A highly significant linear relationship of number of risk factors with heavy alcohol use was found for both sexes. CONCLUSIONS: The rates for men were similar to those reported in a Russian national survey with the exception of Southeast Ukraine where the rate was >10% higher. The highest rates were among men who were middle aged, fathers and unemployed. Future prospective studies are needed to assess the impact of heavy alcohol use on Ukrainian health, mental health and occupational and social functioning. PMID- 15824066 TI - Novel simian immunodeficiency virus CTL epitopes restricted by MHC class I molecule Mamu-B*01 are highly conserved for long term in DNA/MVA-vaccinated, SHIV challenged rhesus macaques. AB - Simian immunodeficiency virus (SIV) infection of rhesus macaques provides an excellent model for investigating the basis of protective immunity against human immunodeficiency virus (HIV). One limitation of this model, however, has been the availability of a small number of known MHC class I-restricted CTL epitopes for investigating virus-specific immune responses. We assessed CTL responses against SIV Gag in a cohort of DNA/modified vaccinia virus Ankara (MVA)-vaccinated/simian human immunodeficiency virus (SHIV)-challenged rhesus macaques. Here, we report the identification of five novel SIV CTL epitopes in Gag for the first time (Gag(39-46) NELDRFGL, Gag(169-177) EVVPGFQAL, Gag(198-206) AAMQIIRDI, Gag(257 265) IPVGNIYRR and Gag(296-305) SYVDRFYKSL) that are restricted by the common MHC class I molecule Mamu-B*01. CTL responses to these epitopes were readily detected in cryopreserved PBMC in multiple animals up to 62 weeks post-infection, both by IFN-gamma enzyme-linked immunospot assay and intracellular IFN-gamma staining. Importantly, viral sequencing results revealed that these epitopes are highly conserved in the SIV-challenged macaques over a long period of time, indicating functional constraints in these regions. Moreover, the presence of CTL responses targeting these epitopes has been confirmed in two independent cohorts of rhesus macaques that have been challenged by SHIV or SIV. Our findings provide valuable candidates for poly-epitope vaccines and for long-term quantitative monitoring of epitope-specific CD8(+) responses in the context of this common Mamu class I allele. It may thus help increase the supply of rhesus macaques in which epitope specific immunity can be studied in the context of SIV vaccine design. PMID- 15824067 TI - Optimizing the exogenous antigen loading of monocyte-derived dendritic cells. AB - Dendritic cell (DC) vaccination, i.e. the adoptive transfer of antigen-loaded DC, is still at an early stage and requires standardization. In this study, we investigated the exogenous loading of monocyte-derived DCs with HLA class I- and II-restricted peptides, as despite widespread use, little effort has been put into its pre-clinical validation. We found that only mature DCs (m-DC) but not immature DCs (im-DC) could be sufficiently loaded with exogenous class I restricted peptides and were by far superior in expanding CD8(+) primary (Melan A.A2 peptide-specific) and recall [Influenza matrix peptide (IMP) A2-specific] T cell responses. Primary stimulation with peptide-loaded im-DCs even down regulated antigen-specific T cell responses. Our results indicate that stimulation with m-DCs is superior in terms of quantity and quality compared with im-DCs, supporting their preferred use in clinical DC trials. Loading of m-DCs with high (10 microM) concentrations generated clearly more Melan-A effectors than loading with 1 or 0.1 microM without any negative effect on the quality (affinity) of the resulting T cells. In contrast to the findings with the Melan-A peptide loading with 10 microM IMP was counter-productive, induced apoptosis and yielded fewer specific T cells of inferior affinity as compared with loading with 1 or 0.1 microM. In sharp contrast to the situation for HLA class I, much higher levels and longer half-lives of peptide-HLA class II complexes were obtainable upon loading of im-DCs with exogenous peptide, but m-DCs were functionally preferable to induce T(h)1 responses in vitro. Another surprising finding was that, while presentation to T cells upon simultaneous loading of several peptides with highly varying affinities and competing for the same class I or II molecule was possible, in priming experiments peptide competition clearly inhibited T cell induction. Although peptides will obviously vary in their individual properties, our study clearly points to some important principles that should be taken into account. PMID- 15824068 TI - Two waves of memory B-cell generation in the primary immune response. AB - Memory B cells can be generated independently of germinal center (GC) formation and affinity maturation in Bcl-6-deficient mice, but the contribution of the GC independent pathway for memory B-cell generation in normal mice remains unknown. To examine this, we administrated anti-inducible co-stimulator (ICOS) mAbs into mice at the onset of GC formation in the primary response. This manipulation affected the generation of GC B cells in the spleen, but neither IgG1 memory B cell nor production of IgG1 long-term antibody was affected. In ICOS-manipulated mice, GC B cells accumulated somatic mutations in the IgV(H) genes and underwent affinity maturation; however, memory B cells scarcely accumulated mutations and reconstituted the secondary response by low affinity, supporting the notion that low-affinity memory B cells are generated in a GC-independent manner. Thus, it appears that memory B cells are established by two different pathways, associated with or without GC reaction and affinity maturation. The generation and long-term persistence of low-affinity IgG1 memory B cells and antibodies in ICOS manipulated mice support the idea that low-affinity memory B cells may give rise to long-term antibody-forming cells. PMID- 15824069 TI - Neutralization of chemokines RANTES and MIG increases virus antigen expression and spinal cord pathology during Theiler's virus infection. AB - The role of chemokines during some viral infections is unpredictable because the inflammatory response regulated by these molecules can have two, contrasting effects-viral immunity and immunopathologic injury to host tissues. Using Theiler's virus infection of SJL mice as a model of this type of disease, we have investigated the roles of two chemokines-regulated on activation, normal T cell expressed and secreted (RANTES) chemokine and monokine induced by IFN-gamma (MIG) by treating mice with antisera that block lymphocyte migration. Control, infected mice showed virus persistence, mild inflammation and a small degree of demyelination in the white matter of the spinal cord at 6 weeks post-infection. Treatment of mice with RANTES antiserum starting at 2 weeks post-infection increased both viral antigen expression and the severity of inflammatory demyelination at 6 weeks post-infection. MIG antiserum increased the spread of virus and the proportion of spinal cord white matter with demyelination. Overall, viral antigen levels correlated strongly with the extent of pathology. At the RNA level, high virus expression was associated with low IL-2 and high IL-10 levels, and RANTES antiserum decreased the IL-2/IL-10 ratio. Our results suggest that RANTES and MIG participate in an immune response that attempts to restrict viral expression while limiting immunopathology and that anti-chemokine treatment poses the risk of exacerbating both conditions in the long term. PMID- 15824070 TI - TGF-beta signaling regulates CD8+ T cell responses to high- and low-affinity TCR interactions. AB - Absence of transforming growth factor-beta (TGF-beta) signaling to T cells in mice results in an increase in T cell numbers, an activated CD44 high, CD69-, CD25- T cell phenotype and a T cell-mediated injury to many organs. It is not known if such T cell activation in the absence of TGF-beta signaling is spontaneous or due to aberrant T cell responses to a physiological stimulus. We used adoptive transfer of CD8+ T cells from mice double transgenic for the OT-1 TCR and the TGF-beta1-dominant negative transgene [OT-dominant-negative receptor (DNR)] to investigate the role of TGF-beta in regulating CD8+ T cell activation in vivo. The activation and expansion of single-transgenic OT and double transgenic OT-DNR cells to oral antigens, high-affinity and low-affinity peptides were indistinguishable. Activation with high-affinity peptide and CFA however resulted in greater expansion of OT-DNR cells in comparison to OT cells. Low affinity peptide and adjuvant did not result in OT cell activation or expansion but results in up-regulation of CD44 on OT-DNR cells. These data show that TGF beta functions in vivo to limit the scale of CD8+ T cell expansion after high affinity peptide-MHC interactions. TGF-beta also limits T cell activation to the highest affinity peptide-MHC interactions. The increase in T cell number and activation present in TGF-beta-deficient and TGF-beta DNR-expressing mice may be due to the loss of these two phenomena. PMID- 15824071 TI - Grafting between model legumes demonstrates roles for roots and shoots in determining nodule type and host/rhizobia specificity. AB - Previous grafting experiments have demonstrated that legume shoots play a critical role in symbiotic development of nitrogen-fixing root nodules by regulating nodule number. Here, reciprocal grafting experiments between the model legumes Lotus japonicus and Medicago truncatula were carried out to investigate the role of the shoot in the host-specificity of legume-rhizobia symbiosis and nodule type. Lotus japonicus is nodulated by Mesorhizobium loti and makes determinate nodules, whereas M. truncatula is nodulated by Sinorhizobium meliloti and makes indeterminate nodules. When inoculated with M. loti, L. japonicus roots grafted on M. truncatula shoots produced determinate nodules identical in appearance to those produced on L. japonicus self-grafted roots. Moreover, the hypernodulation phenotype of L. japonicus har1-1 roots grafted on wild-type M. truncatula shoots was restored to wild type when nodulated with M. loti. Thus, L. japonicus shoots appeared to be interchangeable with M. truncatula shoots in the L. japonicus root/M. loti symbiosis. However, M. truncatula roots grafted on L. japonicus shoots failed to induce nodules after inoculation with S. meliloti or a mixture of S. meliloti and M. loti. Instead, only early responses to S. meliloti such as root hair tip swelling and deformation, plus induction of the early nodulation reporter gene MtENOD11:GUS were observed. The results indicate that the L. japonicus shoot does not support normal symbiosis between the M. truncatula root and its microsymbiont S. meliloti, suggesting that an unidentified shoot-derived factor may be required for symbiotic progression in indeterminate nodules. PMID- 15824072 TI - A role for phosphorylation in the regulation of the barley scutellar peptide transporter HvPTR1 by amino acids. AB - Protein reserves in the cereal endosperm are sequentially degraded to small peptides and amino acids during germination and these are translocated across the scutellum to support growth of the embryo. Peptide transport in the germinating barley grain is mediated by specific carriers localized to the plasma membrane of the scutellar epithelium. In isolated barley embryos peptide transport is rapidly inhibited by amino acid concentrations comparable with those found in the post germination barley grain. However, this inhibition of HvPTR1 activity is not effected at either the transcriptional or translational level. The protein phosphatase inhibitor okadaic acid repressed transport of Ala-[14C]Phe, but not [14C]Ala, into the barley scutellar epithelium. In vivo [32P]orthophosphate labelling studies of barley scutellar tissue in combination with immunoprecipitation studies using antiserum raised to HvPTR1 showed that HvPTR1 (66 kDa) is phosphorylated in the presence of amino acids. Immunopurified HvPTR1 was further demonstrated to be phosphorylated on serine residues. Digestion with the N-glycosidase enzyme PNGase F results in a shift in the molecular mass of the protein by 10 kDa, indicating that HvPTR1 is an N-linked glycoprotein. These results provide strong circumstantial evidence that HvPTR1 peptide transport activity in the germinating barley grain is regulated at the post-translational level by phosphorylation in response to rising levels of amino acids emanating from the endosperm as a result of storage protein breakdown and mobilization. This is potentially an important element in balancing the flux of organic nitrogen and carbon from the endosperm to embryo during germination and seedling establishment. PMID- 15824073 TI - Root-synthesized cytokinin in Arabidopsis is distributed in the shoot by the transpiration stream. AB - To clarify how root-synthesized cytokinins (CKs) are transported to young shoot organs, CK distribution patterns were analysed in free-CK-responsive ARR5::GUS transformants of Arabidopsis thaliana (L.) Heynh. together with free plus bound CKs using specific CK monoclonal antibodies. Plants were subjected to two different growth conditions, completely protected from any air movement, or exposed to gentle wind 3 h before harvesting. In wind-protected plants the strongest ARR5::GUS expression was found in the root cap statocytes, spreading upwards in the vascular cylinder. This pattern in roots was congruent with that found by CK immunolocalization. Shoots of wind-protected plants displayed either no or only low ARR5::GUS expression in the stem vascular bundles, nodal ramifications, and the bases of flower buds; shoot vascular bundles showed patterns of acropetally decreasing staining and the apical parts of buds and leaves were free from ARR5::GUS expression. In wind-exposed plants ARR5::GUS expression was considerably increased in shoots, also in basal-to-apical decreasing gradients. Immunolabelled shoots showed differential staining, with the strongest label in the vascular bundles of stems, leaves, and buds. The fact of the apparent absence of free CK in the buds of wind-protected plants and the typical upward decreasing gradients of free and conjugated CKs suggest that the bulk of the CK is synthesized in the root cap, exported through the xylem and accumulates at sites of highest transpiration where cuticles do not yet exist or do not protect against water loss. PMID- 15824074 TI - The role of alternative oxidase in modulating carbon use efficiency and growth during macronutrient stress in tobacco cells. AB - When wild-type (wt) tobacco (Nicotiana tabacum cv. Petit Havana SR1) cells are grown under macronutrient (P or N) limitation, they induce large amounts of alternative oxidase (AOX), which constitutes a non-energy-conserving branch of the respiratory electron transport chain. To investigate the significance of AOX induction, wt cells were compared with transgenic (AS8) cells lacking AOX. Under nutrient limitation, growth of wt cell cultures was dramatically reduced and carbon use efficiency (g cell dry weight gain g(-1) sugar consumed) decreased by 42-63%. However, the growth of AS8 was only moderately reduced by the nutrient deficiencies and carbon use efficiency values remained the same as under nutrient sufficient conditions. As a result, the nutrient limitations more severely compromised the tissue nutrient status (P or N) of AS8 than wt cells. Northern analyses and a comparison of the mitochondrial protein profiles of wt and AS8 cells indicated that the lack of AOX in AS8 under P limitation was associated with increased levels of proteins commonly associated with oxidative stress and/or stress injury. Also, the level of electron transport chain components was consistently reduced in AS8 while tricarboxylic acid cycle enzymes did not show a universal trend in abundance in comparison to the wt. Alternatively, the lack of AOX in AS8 cells under N limitation resulted in enhanced carbohydrate accumulation. It is concluded that AOX respiration provides an important general mechanism by which plant cells can modulate their growth in response to nutrient availability and that AOX also has nutrient-specific roles in maintaining cellular redox and carbon balance. PMID- 15824075 TI - Carotid stiffness and the risk of new vascular events in patients with manifest cardiovascular disease. The SMART study. AB - AIMS: To study whether arterial stiffness is related to risk of new vascular events in patients with manifest arterial disease and to examine whether this relation varies between patients who differ with respect to baseline vascular risk, arterial stiffness, or systolic blood pressure (SBP). METHODS AND RESULTS: The study was performed in the first consecutive 2183 patients with manifest arterial disease enrolled in the SMART study (Second Manifestations of ARTerial disease), a cohort study among patients with manifest arterial disease or cardiovascular risk factors. Common carotid distension (i.e. the change in carotid diameter in systole relative to diastole) was measured at baseline by ultrasonography. With the distension, several stiffness parameters were determined. In the entire cohort, none of the carotid artery stiffness parameters was related to the occurrence of vascular events. However, decreased stiffness was related to decreased vascular risk in subjects with low baseline SBP. The relation of carotid stiffness with vascular events did not differ between tertiles of baseline risk and carotid stiffness. CONCLUSION: Carotid artery stiffness is no independent risk factor for vascular events in patients with manifest arterial disease. However, in patients with low SBP, decreased carotid stiffness may indicate a decreased risk of vascular events. PMID- 15824076 TI - Complete myocardial revascularization: between myth and reality. AB - Myocardial revascularization in patients with multi-vessel coronary artery disease may be accomplished, by percutaneous interventions or surgery, either on all diseased lesions or directed to selectively targeted coronary segments. The extent of planned revascularization is often a major determinant of treatment strategy. Revascularization of all diseased coronary segments-complete myocardial revascularization-has a potential long-term benefit, but is more complex and may increase in-hospital untoward events. Revascularization may otherwise be incomplete, either because of the operator's inability to treat all diseased coronary segments or by choice of deciding to selectively revascularize only large areas of myocardium at risk. Although incomplete revascularization may negatively affect long-term outcomes, it may be, when wisely chosen, the preferred treatment strategy in selected patient categories because of its lower immediate risks. The patient's clinical status, ventricular function, and the presence of co-morbidities may orient clinical decisions in favour of incomplete revascularization. PMID- 15824077 TI - Lipoprotein (a) and coronary heart disease among women: beyond a cholesterol carrier? AB - AIMS: With its homology with plasminogen, lipoprotein(a) [Lp(a)] may be related to thrombosis and inflammation. We assessed the role of Lp(a) in coronary heart diseases (CHD) by a recently developed assay that is not affected by the plasminogen-like Kringle-type-2 repeats. METHODS AND RESULTS: Of 32 826 women from the Nurses' Health Study, who provided blood at baseline, we documented 228 CHD events during 8 years of follow-up. Each case was compared with two matched controls. In a multivariable model adjusted for body mass index, family history, hypertension, diabetes, post-menopausal hormone use, physical activity, blood drawing characteristics, and alcohol intake, the odd ratio (OR) for Lp(a) levels > or =30 mg/dL was 1.9(95% CI: 1.3-3.0) when compared with those with Lp(a)<30 mg/dL. Women with high levels of both Lp(a) (> or =30 mg/dL) and fibrinogen (> or =400 mg/dL) had an OR of 3.2(95% CI: 1.6-6.5) for CHD, when compared with the combination of low levels (P interaction=0.05). Women with high levels of both Lp(a) and C-reactive protein (> or =3 mg/L) had an OR of 3.67(95% CI: 2.03-6.64) for CHD, when compared with the combination of low levels (P interaction=0.06). CONCLUSION: Lp(a) levels >30 mg/dL are associated with twice the risk of CHD events among women and may be related to thrombosis and inflammation. PMID- 15824078 TI - A dynamic model forecasting myocardial infarct size before, during, and after reperfusion therapy: an ASSENT-2 ECG/VCG substudy. AB - AIMS: Serial forecasts of final myocardial infarct (MI) size during fibrinolytic treatment (Rx) of ST-elevation MI would allow the identification of high-risk patients with a predicted major loss of viable myocardium, at a point when treatment may still be modified. We investigated a model for such forecasting, using time and the ECG. METHODS AND RESULTS: We collected 234 patients with ST elevation MI, without signs of previous MI, bundle branch block, or hypertrophy. MI size was determined by the Selvester score and was "forecasted" at: admission with patients stratified by delay time and an ECG acuteness score into three groups (EARLY, DISCORDANT, and LATE); 90 min after Rx by > or =70% ST-recovery or not and occurrence of "reperfusion peaks"; 4 h after Rx by ST re-elevations. EARLY patients had smaller final infarct sizes than LATE (9.4 vs. 20%, P=0.01). EARLY patients with > or =70% ST-recovery without a reperfusion peak had smaller infarct sizes than those with (3.1 vs. 12.5%, P=0.001). EARLY patients without ST re-elevations had smaller infarct sizes (1.5%) than those with some (9%) or many re-elevations (12%), P<0.001. CONCLUSION: Final infarct size can be forecasted using delay time and serial ECGs. Serially updated forecasts seem especially important when both clock-time and initial ECG- signs indicate earliness. PMID- 15824079 TI - Impedance compensated biphasic waveforms for transthoracic cardioversion of atrial fibrillation: a multi-centre comparison of antero-apical and antero posterior pad positions. AB - AIMS: To compare the success rate for transthoracic direct current cardioversion (DCC) of atrial fibrillation (AF) with antero-posterior (AP) and antero-apical (AA) electrode positions using an impedance compensated biphasic (ICB) waveform. METHODS AND RESULTS: Three-hundred and seven patients [mean age 66 (SD+/-13), 195 male] with AF were recruited in three centres. Patients were randomized to an AA (n=150) or AP (n=144) pad position. Thirteen patients with implanted pacemakers were defaulted to the AP pad position. Cardioversion was performed using an ICB waveform with a 70, 100, 150, and 200 J energy selection protocol. If the fourth shock was unsuccessful, the pads were crossed over to the alternative position for a final 200 J shock. Shock 1 was successful in 54/150 (36%) AA and 45/144 (31%) AP patients, whereas success was achieved by shock 2 in 99/150 (66%) AA and 74/144 (51%) AP, by shock 3 in 123/150 (82%) AA and 109/144 (76%) AP, and by shock 4 in 143/150 (95%) AA and 127/144 (88%) AP and after cross-over in 144/150 (96%) AA and 135/144 (94%) AP. Overall success rate was higher than expected at 95%. Pad position was not associated significantly with success. There was a trend towards an improved outcome with the AA configuration (P=0.05). CONCLUSION: The influence of pad position for DCC of AF may be less pertinent with ICB waveforms than with monophasic waveforms. PMID- 15824080 TI - Second-line (post-study) chemotherapy received by patients treated in the phase III trial of pemetrexed plus cisplatin versus cisplatin alone in malignant pleural mesothelioma. AB - BACKGROUND: A phase III trial in patients with malignant pleural mesothelioma demonstrated a survival advantage for pemetrexed plus cisplatin compared with single-agent cisplatin. Because post-study chemotherapy (PSC) may have influenced the outcome of the trial, we examined its use and association with survival. PATIENTS AND METHODS: Eighty-four patients from the pemetrexed plus cisplatin arm and 105 patients from the single-agent cisplatin arm received PSC. Kaplan-Meier survival estimates were compared by treatment groups, and by PSC and non-PSC subgroups. RESULTS: The percentage of patients receiving PSC was imbalanced between the treatment arms. Fewer pemetrexed plus cisplatin treated patients received PSC (37.2% versus 47.3%). A multiple regression analysis performed in this trial showed that PSC had a statistically significant correlation with prolonged survival (P <0.01), adjusting for baseline prognostic factors and treatment intervention. The adjusted hazard ratio for PSC over non-PSC subgroups was 0.56 (confidence interval 0.44-0.72). CONCLUSIONS: PSC in malignant pleural mesothelioma was significantly associated with prolonged survival. It is not known whether the reduced risk of death was associated with PSC or whether patients who had prolonged survival tended to receive more PSC. The pemetrexed plus cisplatin treatment group had a statistically significant survival advantage even though fewer patients from that arm of the trial received PSC. The potentially beneficial role of PSC should be assessed in prospective trials. PMID- 15824081 TI - A phase I study of antisense oligonucleotide GTI-2040 given by continuous intravenous infusion in patients with advanced solid tumors. AB - BACKGROUND: This study of GTI-2040, a 20-mer phosphorothioate oligonucleotide complementary to the messenger ribonucleic acid (mRNA) of the R2 subunit of ribonucleotide reductase (RNR), was conducted to determine the dose-limiting toxicity (DLT) and maximum-tolerated dose (MTD) of the agent in patients with advanced solid tumors or lymphoma. Plasma pharmacokinetics of GTI-2040 and suppression of RNR expression in peripheral blood mononuclear cells were also studied. PATIENTS AND METHODS: GTI-2040 was administered as a continuous intravenous infusion for 21 days every 4 weeks. Dose escalation was performed using an accelerated, dose-doubling schedule until any drug related toxicity > or = grade 2 was observed; subsequent dose escalation followed a more conservative dose escalation scheme with three patients/cohort. RESULTS: A total of 49 cycles of therapy were administered to 36 patients at GTI-2040 doses ranging from 18.5 mg/m(2)/day to 222 mg/m(2)/day. GTI-2040 was generally well tolerated. At the highest dose level examined, two patients experienced dose limiting reversible hepatic toxicity. Constitutional toxicities consisting of fatigue and anorexia were the most common toxicities. CONCLUSIONS: The recommended dose of GTI-2040 given on this infusion schedule is 185 mg/m(2)/day. GTI-2040 appears to have a manageable toxicity profile and is generally well tolerated as a single agent. PMID- 15824082 TI - Protection from angiotensin II-mediated vasculotoxic and hypertensive response in mice lacking PI3Kgamma. AB - Hypertension affects nearly 20% of the population in Western countries and strongly increases the risk for cardiovascular diseases. In the pathogenesis of hypertension, the vasoactive peptide of the renin-angiotensin system, angiotensin II and its G protein-coupled receptors (GPCRs), play a crucial role by eliciting reactive oxygen species (ROS) and mediating vessel contractility. Here we show that mice lacking the GPCR-activated phosphoinositide 3-kinase (PI3K)gamma are protected from hypertension that is induced by administration of angiotensin II in vivo. PI3Kgamma was found to play a role in angiotensin II-evoked smooth muscle contraction in two crucial, distinct signaling pathways. In response to angiotensin II, PI3Kgamma was required for the activation of Rac and the subsequent triggering of ROS production. Conversely, PI3Kgamma was necessary to activate protein kinase B/Akt, which, in turn, enhanced L-type Ca(2+) channel mediated extracellular Ca(2+) entry. These data indicate that PI3Kgamma is a key transducer of the intracellular signals that are evoked by angiotensin II and suggest that blocking PI3Kgamma function might be exploited to improve therapeutic intervention on hypertension. PMID- 15824083 TI - Intestinal antiinflammatory effect of 5-aminosalicylic acid is dependent on peroxisome proliferator-activated receptor-gamma. AB - 5-aminosalicylic acid (5-ASA) is an antiinflammatory drug widely used in the treatment of inflammatory bowel diseases. It is known to inhibit the production of cytokines and inflammatory mediators, but the mechanism underlying the intestinal effects of 5-ASA remains unknown. Based on the common activities of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligands and 5-ASA, we hypothesized that this nuclear receptor mediates 5-ASA therapeutic action. To test this possibility, colitis was induced in heterozygous PPAR-gamma(+/-) mice and their wild-type littermates, which were then treated with 5-ASA. 5-ASA treatment had a beneficial effect on colitis only in wild-type and not in heterozygous mice. In epithelial cells, 5-ASA increased PPAR-gamma expression, promoted its translocation from the cytoplasm to the nucleus, and induced a modification of its conformation permitting the recruitment of coactivators and the activation of a peroxisome-proliferator response element-driven gene. Validation of these results was obtained with organ cultures of human colonic biopsies. These data identify PPAR-gamma as a target of 5-ASA underlying antiinflammatory effects in the colon. PMID- 15824084 TI - CD44 is a physiological E-selectin ligand on neutrophils. AB - The selectin family of adhesion molecules and their glycoconjugated ligands are essential for blood polymorphonuclear neutrophil (PMN) extravasation into inflammatory and infectious sites. However, E-selectin ligands on PMNs are not well characterized. We show here that CD44 immunopurified from G-CSF differentiated 32D cells or from peripheral blood PMNs binds specifically to E selectin. In contrast, CD44 extracted from bone marrow stromal or brain endothelial cell lines does not interact with E-selectin, suggesting cell specific posttranslational modifications of CD44. PMN-derived CD44 binding activity is mediated by sialylated, alpha(1,3) fucosylated, N-linked glycans. CD44 enables slow leukocyte rolling on E-selectin expressed on inflamed endothelium in vivo and cooperates with P-selectin glycoprotein ligand-1 to recruit neutrophils into thioglycollate-induced peritonitis and staphylococcal enterotoxin A-injected skin pouch. CD44 extracted from human PMNs also binds to E selectin. Moreover, we demonstrate that CD44 is hypofucosylated in PMNs from a patient with leukocyte adhesion deficiency type II, suggesting that it contributes to the syndrome. These findings thus suggest broader roles for CD44 in the innate immune response and uncover a potential new target for diseases in which selectins play a prominent role. PMID- 15824085 TI - Boosting antitumor responses of T lymphocytes infiltrating human prostate cancers. AB - Immunotherapy may provide valid alternative therapy for patients with hormone refractory metastatic prostate cancer. However, if the tumor environment exerts a suppressive action on antigen-specific tumor-infiltrating lymphocytes (TIL), immunotherapy will achieve little, if any, success. In this study, we analyzed the modulation of TIL responses by the tumor environment using collagen gel matrix-supported organ cultures of human prostate carcinomas. Our results indicate that human prostatic adenocarcinomas are infiltrated by terminally differentiated cytotoxic T lymphocytes that are, however, in an unresponsive status. We demonstrate the presence of high levels of nitrotyrosines in prostatic TIL, suggesting a local production of peroxynitrites. By inhibiting the activity of arginase and nitric oxide synthase, key enzymes of L-arginine metabolism that are highly expressed in malignant but not in normal prostates, reduced tyrosine nitration and restoration of TIL responsiveness to tumor were achieved. The metabolic control exerted by the tumor on TIL function was confirmed in a transgenic mouse prostate model, which exhibits similarities with human prostate cancer. These results identify a novel and dominant mechanism by which cancers induce immunosuppression in situ and suggest novel strategies for tumor immunotherapy. PMID- 15824086 TI - Contribution of DNA polymerase eta to immunoglobulin gene hypermutation in the mouse. AB - The mutation pattern of immunoglobulin genes was studied in mice deficient for DNA polymerase eta, a translesional polymerase whose inactivation is responsible for the xeroderma pigmentosum variant (XP-V) syndrome in humans. Mutations show an 85% G/C biased pattern, similar to that reported for XP-V patients. Breeding these mice with animals harboring the stop codon mutation of the 129/Olain background in their DNA polymerase iota gene did not alter this pattern further. Although this G/C biased mutation profile resembles that of mice deficient in the MSH2 or MSH6 components of the mismatch repair complex, the residual A/T mutagenesis of pol eta-deficient mice differs markedly. This suggests that, in the absence of pol eta, the MSH2-MSH6 complex is able to recruit another DNA polymerase that is more accurate at copying A/T bases, possibly pol kappa, to assume its function in hypermutation. PMID- 15824087 TI - FOXO-dependent expression of the proapoptotic protein Bim: pivotal role for apoptosis signaling in endothelial progenitor cells. AB - Endothelial progenitor cells (EPCs) contribute to postnatal neovascularization. Risk factors for coronary artery disease reduce the number of EPCs in humans. Since EPC apoptosis might be a potential mechanism to regulate the number of EPCs, we investigated the effects of oxidative stress and HMG-CoA-reductase inhibitors (statins) on EPC apoptosis. Atorvastatin, mevastatin, or VEGF prevented EPC apoptosis induced by H2O2. The antiapoptotic effect was reversed by inhibition of the PI3K/Akt pathway. Forkhead transcription factors (FOXO1, FOXO3a, FOXO4) exert proapoptotic effects and are phosphorylated and, thereby, inactivated by Akt. Therefore, we elucidated the involvement of forkhead transcription factors. Atorvastatin induced the phosphorylation of the predominant forkhead factor FOXO4 in EPCs. In addition, atorvastatin reduced the expression of the proapoptotic forkhead-regulated protein Bim in a PI3K-dependent manner. Consistently, overexpression of FOXO4 activated the Bim promoter as determined by reporter gene expression and stimulated the expression of Bim, resulting in an increased EPC apoptosis. Statins failed to prevent EPC apoptosis induced by overexpression of Bim or nonphosphorylatable FOXO4, suggesting that the protective effects of statins depend on this pathway. In summary, our results show that FOXO-dependent expression of Bim plays a pivotal role for EPC apoptosis. Statins reduce oxidative stress-induced EPC apoptosis, inactivate FOXO4, and down-regulate Bim. PMID- 15824088 TI - Heteroresistance to carbapenems in Acinetobacter baumannii. PMID- 15824089 TI - Use of lopinavir/ritonavir in HIV-infected patients failing a first-line protease inhibitor-containing HAART. AB - OBJECTIVES: The long-term virological efficacy of lopinavir/ritonavir-containing highly active antiretroviral therapy (HAART) in HIV-infected patients failing a first-line protease inhibitor (PI)-based regimen is still unclear. METHODS: An observational study was carried out from December 2000-December 2002 on 111 consecutive patients starting lopinavir/ritonavir. The primary end-point was virological success (HIV RNA <50 copies/mL in two consecutive determinations). CD4 outcome, lipid levels and adverse events were recorded. The Kaplan-Meier method and log-rank test were used to estimate the time-dependent probability of reaching the end-point using intention-to-treat and on-treatment approaches. RESULTS: Ninety-six patients obtained virological success during follow-up; Kaplan-Meier analysis showed that the time-dependent probability of obtaining this end-point was 78.4% at month 12 and 85.8% at month 24. The median CD4+cell count increased by 118 cells/mm(3) from baseline to month 12 and by 153 cells/mm(3) to month 24. Thirty-one patients discontinued lopinavir/ritonavir: 16 because of drug-related toxicities, six for simplification, five because of virological failure, one patient was lost at follow-up and three died. An elevation in lipid parameters was observed, but only a minority of patients developed a grade 3 or higher hypertriglyceridaemia and/or hypercholesterolaemia. Among the 15 patients not reaching virological success, five had < or =5 mutations in the protease region known to reduce susceptibility to lopinavir/ritonavir (one discontinued lopinavir/ritonavir because of gastrointestinal intolerance), five had no mutations (two discontinued lopinavir/ritonavir because of gastrointestinal intolerance) and five showed > or =6 mutations (all discontinued lopinavir/ritonavir); however, of the patients who discontinued lopinavir/ritonavir none achieved HIV RNA <50 copies/mL on subsequent regimens. CONCLUSIONS: Lopinavir/ritonavir was highly effective and well tolerated in HIV-infected patients failing a first-line PI-based HAART. PMID- 15824090 TI - In vitro activity of linezolid, clarithromycin and moxifloxacin against clinical isolates of Mycobacterium kansasii. AB - OBJECTIVES: To compare the activity of linezolid with a range of drugs used in the treatment of Mycobacterium kansasii infections. RESULTS: The percentages of resistant isolates against isoniazid, rifampicin and ethambutol were 2.9%, 1.9% and 2.9%, respectively. All isolates were susceptible to clarithromycin and moxifloxacin both with MIC(90) values of 0.125 mg/L. Linezolid was active against all isolates with MIC(50) and MIC(90) values of 0.5 and 1 mg/L, respectively, both below the susceptibility breakpoint established for mycobacteria. CONCLUSION: Linezolid, clarithromycin or moxifloxacin, could be used as alternative drugs for treatment of infections due to rifampicin-resistant isolates as well as short-course or intermittent therapy of M. kansasii lung disease. PMID- 15824091 TI - Propyl paraben induces potassium efflux in Escherichia coli. AB - OBJECTIVES: Parabens are currently used as antibacterial preservatives in pharmaceutical, cosmetic and food products but there are no precise data concerning their activity on bacterial membranes. METHODS: We analysed the cytoplasmic potassium release during propyl paraben addition by using a selective electrode. Various conditions were assayed to investigate the bacterial paraben susceptibility. We compared the activity of propyl paraben with the activities of colicin A and polymyxin B. RESULTS: Propyl paraben induced potassium efflux that was related to the porin expression in the bacterial outer membrane. In addition, the presence of spermine, previously described as an efficient OmpF channel blocker, protected susceptible cells against paraben activity. CONCLUSIONS: Propyl paraben induced potassium release in susceptible Escherichia coli cells similar to that observed with polymyxin B. Moreover, this efflux depended on porin channel activity. This permeabilizing effect is probably related to antibacterial properties of paraben molecules. PMID- 15824092 TI - Evaluation of the in vitro activity of NVP-LMB415 against clinical anaerobic isolates with emphasis on the Bacteroides fragilis group. AB - OBJECTIVES: To compare the in vitro activity of NVP-LMB415 (formerly referred to as NVP-PDF 713) with that of other agents with anti-anaerobe activity against clinical anaerobic isolates, with emphasis on the Bacteroides fragilis group. METHODS: The MICs for 405 B. fragilis group and 102 Gram-positive anaerobic isolates were determined using NCCLS-recommended procedures. The activity of NVP LMB415 was compared with that of cefoxitin, clindamycin, imipenem, garenoxacin, linezolid, moxifloxacin and tigecycline. Vancomycin was included in the evaluation of the Gram-positive organisms. RESULTS: NVP-LMB415 showed excellent in vitro activity against all the species of the B. fragilis group isolates (MIC range < or = 0.03-0.5 mg/L and MIC(90) 0.5 mg/L). NVP-LMB415 was active against B. fragilis group strains resistant to beta-lactams, quinolones or clindamycin, and the MICs were much lower than those of newer agents such as linezolid, tigecycline and garenoxacin. The MICs of NVP-LMB415 ( > or = 4 mg/L) for Clostridium species were higher than the MICs for other anaerobes. CONCLUSIONS: Given the frequency of isolation of anaerobic bacteria and their increasing resistance to all classes of antibiotics, NVP-LMB415 is an ideal agent for potential use against mixed infections caused by resistant anaerobic pathogens such as of B. fragilis and Gram-positive aerobic strains such as methicillin resistant staphylococci, streptococci and enterococci. PMID- 15824093 TI - A comparative study of fungicidal activities of voriconazole and amphotericin B against hyphae of Aspergillus fumigatus. AB - OBJECTIVES: To study the in vitro fungicidal activity of voriconazole against hyphae of Aspergillus fumigatus and compare the results with those obtained for the known fungicidal drug amphotericin B. METHODS: A. fumigatus mycelia were grown on Sabouraud dextrose agar and in peptone yeast extract glucose broth until the cultures reached a mid-logarithmic growth phase. The fungicidal activities of voriconazole and amphotericin B against actively growing hyphae of A. fumigatus were examined by a kill-curve experiment and a fungal cell viability test. For the kill-curve study, the drug-treated hyphae were washed, homogenized and resuspended in 1 mL of sterile water, diluted 10-1000 fold and aliquots of 0.1 mL were spread on Sabouraud dextrose agar and allowed to grow for 48 h at 35 degrees C. The cfu were determined and plotted against drug concentrations for each time of exposure to obtain the kill curve. The viability of drug-treated A. fumigatus hyphae was determined by their ability to reduce tetrazolium compound 3-(4,5 dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide. RESULTS: Exposure of A. fumigatus hyphae to several concentrations (1-16 mg/L) of voriconazole or amphotericin B for various time intervals killed the hyphae in a time- and drug concentration-dependent manner. Voriconazole at 1 mg/L killed >95% of the hyphae grown on Sabouraud dextrose agar after 48 h of exposure, whereas amphotericin B at the same concentration killed approximately 70% of the hyphae after exposure for the same duration. Approximately 99% killing of hyphae grown in peptone yeast extract glucose broth was obtained for voriconazole at 1 mg/L after 48 h of exposure, whereas amphotericin B at 1 mg/L yielded approximately 82% killing after 48 h. The fungal cell viability test by tetrazolium reduction assay showed that mycelia exposed to > or =1 mg/L (Sabouraud dextrose agar blocks) and > or =2 mg/L (broth cultures) of voriconazole for 48 h completely failed to reduce 3-(4,5 dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide. At low concentrations (1-2 mg/L) amphotericin B had no detectable effect on 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction by drug-treated mycelia, whereas mycelia treated with 16 mg/L for 48 h showed approximately 50% inhibition of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction compared with the control. CONCLUSIONS: Voriconazole possesses excellent fungicidal activity against actively growing hyphae of A. fumigatus. A comparison of results with those obtained for the known fungicidal drug amphotericin B shows that, in peptone yeast extract glucose broth, voriconazole has superior fungicidal activity against A. fumigatus hyphae compared with that of amphotericin B. PMID- 15824094 TI - In vitro antiplasmodial activity of prenylated chalcone derivatives of hops (Humulus lupulus) and their interaction with haemin. AB - OBJECTIVES: There is an urgent need to discover new antimalarials, due to the spread of chloroquine resistance and the limited number of available drugs. Chalcones are one of the classes of natural products that are known to possess antiplasmodial properties. Therefore, the in vitro antiplasmodial activity of the main hop chalcone xanthohumol and seven derivatives was evaluated. In addition, the influence of the compounds on glutathione (GSH)-dependent haemin degradation was analysed to determine its contribution to the antimalarial effect of chalcones. METHODS: In vitro antiplasmodial activity was evaluated against the chloroquine-sensitive strain poW and the multiresistant clone Dd2 using a [(3)H]hypoxanthine-incorporation assay. Inhibition of GSH-dependent haemin degradation was analysed by a multiwell plate assay at 11 microM. RESULTS: Of the eight compounds tested, four possessed activity with IC(50) values<25 microM against at least one of the two strains of Plasmodium falciparum. The main hop chalcone, xanthohumol, was most active with IC(50) values of 8.2+/-0.3 (poW) and 24.0 +/- 0.8 microM (Dd2). Three of these compounds were additionally active in the haemin-degradation assay. CONCLUSIONS: The results demonstrate for the first time the ability of chalcone derivatives to interfere with the haemin-degradation process of P. falciparum. This effect might contribute to their antiplasmodial activity. Nevertheless, as one compound showed inhibition of P. falciparum without being able to interact with GSH-dependent haemin degradation, other modes of action must add to the observed antiparasitic activity of hop chalcones. PMID- 15824095 TI - Uptake and intracellular activity of voriconazole in human polymorphonuclear leucocytes. AB - OBJECTIVES: The intracellular penetration of voriconazole into human polymorphonuclear leucocytes (PMNs) and its intracellular activity against Candida spp. were evaluated. METHODS: The intracellular penetration of voriconazole into PMNs was evaluated by a radiometric assay. The effect of cell viability, environmental conditions, metabolic inhibitors and membrane stimulation was also studied. The intracellular activity was determined by incubation of PMNs containing intracellular blastospores in the presence of voriconazole for 3 h. RESULTS: The uptake of voriconazole by PMNs was rapid and not saturable. The cellular to extracellular concentration (C/E) ratio for voriconazole was 8.5+/-1.3. Voriconazole was rapidly released from loaded PMNs. The uptake of voriconazole was not affected by environmental temperature and cell viability. Neither the external pH nor the metabolic inhibitors affected the uptake of voriconazole. The ingestion of opsonized zymosan, but not of opsonized Candida spp., significantly decreased the levels of PMN-associated voriconazole. At the extracellular concentrations evaluated, voriconazole did not affect the intracellular survival of Candida. CONCLUSIONS: Voriconazole reached high intracellular concentrations within human PMNs. The uptake was rapid and not saturable but it did not affect the intracellular killing of Candida spp. PMID- 15824096 TI - Binding of ADAMTS13 to von Willebrand factor. AB - ADAMTS13, a metalloprotease, cleaves von Willebrand factor (VWF) in plasma to generate smaller, less thrombogenic fragments. The interaction of von Willebrand factor with specific ADAMTS13 domains was characterized with a binding assay employing von Willebrand factor immobilized on a plastic surface. ADAMTS13 binding was saturable and reversible. Equilibrium binding occurred within 2 h and the half-time for dissociation was approximately 4 h. Binding to von Willebrand factor was similar with either recombinant ADAMTS13 or normal plasma ADAMTS13; plasma from a patient who lacked ADAMTS13 activity showed no binding. The stoichiometry of binding was one ADAMTS13 per two von Willebrand factor monomers, and the K(d) was 14 nm. The ADAMTS13 metalloprotease and disintegrin domains did not bind VWF detectably. ADAMTS13 truncated after the first thrombospondin type 1 repeat bound VWF with a K(d) of 206 nm, whereas ADAMTS13 truncated after the spacer domain had a K(d) of 23 nm, which is comparable with that of full-length ADAMTS13. Truncation after the eighth thrombospondin type 1 repeat reduced the binding affinity by approximately 3-fold and truncation after the seventh thrombospondin type 1 repeat in addition to the CUB domains increased the affinity for von Willebrand factor by approximately 2-fold. Therefore, the spacer domain is required for ADAMTS13 binding to von Willebrand factor. The first thrombospondin repeat also affects binding, and the C-terminal thrombospondin type 1 and CUB domains of ADAMTS13 may modulate this interaction. PMID- 15824097 TI - Mind bomb-2 is an E3 ligase for Notch ligand. AB - The zebrafish gene, mind bomb (mib), encodes a protein that positively regulates of the Delta-mediated Notch signaling. It interacts with the intracellular domain of Delta to promote its ubiquitination and endocytosis. In our search for the mouse homologue of zebrafish mind bomb, we cloned two homologues in the mouse genome: a mouse orthologue (mouse mib1) and a paralogue, named mind bomb-2 (mib2), which is evolutionarily conserved from Drosophila to human. Both Mib1 and Mib2 have an E3 ubiquitin ligase activity in their C-terminal RING domain and interact with Xenopus Delta (XD) via their N-terminal region. Mib2 is also able to ligate ubiquitin to XD and shift the membrane localization of Delta to intracellular vesicles. Importantly, Mib2 rescues both the neuronal and vascular defects in the zebrafish mib(ta52b) mutants. In contrast to the functional similarities between Mib1 and Mib2, mib2 is highly expressed in adult tissues, but almost not at all in embryos, whereas mib1 is abundantly expressed in both embryos and adult tissues. These data suggest that Mib2 has functional similarities to Mib1, but might have distinct roles in Notch signaling as an E3 ubiquitin ligase. PMID- 15824098 TI - Targeted deletion of the chicken beta-globin regulatory elements reveals a cooperative gene silencing activity. AB - The chicken beta-globin locus represents a well characterized system to study the role of both proximal and distal regulatory elements in a eukaryotic multigene domain. The function of the chicken beta(A)/epsilon-intergenic enhancer and upstream regulatory elements 5'-HS1 and 5'-HS2 were studied using a gene targeting approach in chicken DT40 cells followed by microcell-mediated chromosome transfer into human erythroleukemia cells (K562). These regulatory elements all repressed expression of the rho- and beta(H)-chicken globin genes in the chromosome transfer assay. No rho- or beta(H)-globin gene expression was detected in K562 cells containing the chicken chromosome without deletions, whereas rho- and beta(H)-mRNA was activated in K562 cells containing chicken chromosomes with deletions of the intergenic enhancers, 5'-HS1 and 5'-HS2. Transcriptional activation of the rho- and beta(H)-globin genes correlated with hyperacetylation of histones H3 and H4, loss of histone H3 lysine 9 methylation, and binding of RNA polymerase II to the gene promoters. Surprisingly, the status of CpG dinucleotide methylation at the promoters did not correlate with the transcriptional status of the genes. Our results using a chromosomal transfer assay demonstrate an identical silencing function for these regulatory elements, which suggests they function as part of a common silencing pathway or complex. PMID- 15824099 TI - Detoxification and transcriptome response in Arabidopsis seedlings exposed to the allelochemical benzoxazolin-2(3H)-one. AB - Benzoxazolin-2(3H)-one (BOA) is an allelochemical most commonly associated with monocot species, formed from the O-glucoside of 2,4-dihydroxy-2H-1,4-benzoxazin 3(4H)-one by a two-step degradation process. The capacity of Arabidopsis to detoxify exogenously supplied BOA was analyzed by quantification of the major known metabolites BOA-6-OH, BOA-6-O-glucoside, and glucoside carbamate, revealing that detoxification occurs predominantly through O-glucosylation of the intermediate BOA-6-OH, most likely requiring the sequential action of as-yet unidentified cytochrome P450 and UDP-glucosyltransferase activities. Transcriptional profiling experiments were also performed with Arabidopsis seedlings exposed to BOA concentrations, representing I(50) and I(80) levels based on root elongation inhibition assays. One of the largest functional categories observed for BOA-responsive genes corresponded to protein families known to participate in cell rescue and defense, with the majority of these genes potentially associated with chemical detoxification pathways. Further experiments using a subset of these genes revealed that many are also transcriptionally induced by a variety of structurally diverse xenobiotic compounds, suggesting they comprise components of a coordinately regulated, broad specificity xenobiotic defense response. The data significantly expand upon previous studies examining plant transcriptional responses to allelochemicals and other environmental toxins and provide novel insights into xenobiotic detoxification mechanisms in plants. PMID- 15824100 TI - Modulation of prion-dependent polyglutamine aggregation and toxicity by chaperone proteins in the yeast model. AB - In yeast, aggregation and toxicity of the expanded polyglutamine fragment of human huntingtin strictly depend on the presence of the endogenous self perpetuating aggregated proteins (prions), which contain glutamine/asparagine rich domains. Some chaperones of the Hsp100/70/40 complex, modulating propagation of yeast prions, were also reported to influence polyglutamine aggregation in yeast, but it was not clear whether they do it directly or via affecting prions. Our data show that although some chaperone alterations indeed act on polyglutamines via curing endogenous prions, other alterations decrease size and ameliorate toxicity of polyglutamine aggregates without affecting prion propagation. Therefore, the role of yeast chaperones in polyglutamine aggregation and toxicity is not restricted only to their effects on the endogenous prions. Moreover, chaperone interactions with prion and polyglutamine aggregates appear to be of a highly specific nature. One and the same chaperone alteration, substitution A503V in the middle region of the chaperone Hsp104, exhibited opposite effects on one of the endogenous prions ([PSI(+)], the prion form of Sup35) and on polyglutamines, increasing aggregate size and toxicity in the former case and decreasing them in the latter case. On the other hand, different members of a single chaperone family exhibited opposite effects on one and the same type of aggregates: excess of the Hsp40 chaperone Ydj1 increased polyglutamine aggregate size and toxicity, whereas excess of the other Hsp40 chaperone, Sis1, decreased them. As many stress-defense proteins are conserved between yeast and mammals, these data shed light on possible mechanisms modulating polyglutamine aggregation and toxicity in mammalian cells. PMID- 15824101 TI - Amidation and structure relaxation abolish the neurotoxicity of the prion peptide PrP106-126 in vivo and in vitro. AB - One of the major pathological hallmarks of transmissible spongiform encephalopathies (TSEs) is the accumulation of a pathogenic (scrapie) isoform (PrP(Sc)) of the cellular prion protein (PrP(C)) primarily in the central nervous system. The synthetic prion peptide PrP106-126 shares many characteristics with PrP(Sc) in that it shows PrP(C)-dependent neurotoxicity both in vivo and in vitro. Moreover, PrP106-126 in vitro neurotoxicity has been closely associated with the ability to form fibrils. Here, we studied the in vivo neurotoxicity of molecular variants of PrP106-126 toward retinal neurons using electroretinographic recordings in mice after intraocular injections of the peptides. We found that amidation and structure relaxation of PrP106-126 significantly reduced the neurotoxicity in vivo. This was also found in vitro in primary neuronal cultures from mouse and rat brain. Thioflavin T binding studies showed that amidation and structure relaxation significantly reduced the ability of PrP106-126 to attain fibrillar structures in physiological salt solutions. This study hence supports the assumption that the neurotoxic potential of PrP106 126 is closely related to its ability to attain secondary structure. PMID- 15824102 TI - beta Subunits of voltage-gated sodium channels are novel substrates of beta-site amyloid precursor protein-cleaving enzyme (BACE1) and gamma-secretase. AB - Sequential processing of amyloid precursor protein (APP) by membrane-bound proteases, BACE1 and gamma-secretase, plays a crucial role in the pathogenesis of Alzheimer disease. Much has been discovered on the properties of these proteases; however, regulatory mechanisms of enzyme-substrate interaction in neurons and their involvement in pathological changes are still not fully understood. It is mainly because of the membrane-associated cleavage of these proteases and the lack of information on new substrates processed in a similar way to APP. Here, using RNA interference-mediated BACE1 knockdown, mouse embryonic fibroblasts that are deficient in either BACE1 or presenilins, and BACE1-deficient mouse brain, we show clear evidence that beta subunits of voltage-gated sodium channels are sequentially processed by BACE1 and gamma-secretase. These results may provide new insights into the underlying pathology of Alzheimer disease. PMID- 15824103 TI - The NADPH oxidase Nox3 constitutively produces superoxide in a p22phox-dependent manner: its regulation by oxidase organizers and activators. AB - Nox3, a member of the superoxide-producing NADPH oxidase (Nox) family, participates in otoconia formation in mouse inner ears, which is required for perception of balance and gravity. The activity of other Nox enzymes such as gp91(phox)/Nox2 and Nox1 is known to absolutely require both an organizer protein (p47(phox) or Noxo1) andanactivatorprotein (p67(phox) or Noxa1); for the p47(phox)-dependent activation of these oxidases, treatment of cells with stimulants such as phorbol 12-myristate 13-acetate is also indispensable. Here we show that ectopic expression of Nox3 in various types of cells leads to phorbol 12-myristate 13-acetate-independent constitutive production of a substantial amount of superoxide under the conditions where gp91(phox) and Nox1 fail to generate superoxide, i.e. in the absence of the oxidase organizers and activators. Nox3 likely forms a functional complex with p22(phox); Nox3 physically interacts with and stabilizes p22(phox), and the Nox3-dependent superoxide production is totally dependent on p22(phox). The organizers p47(phox) and Noxo1 are capable of enhancing the superoxide production by Nox3 in the absence of the activators, and the enhancement requires the interaction of the organizers with p22(phox), further indicating a link between Nox3 and p22(phox). The p47(phox)-enhanced Nox3 activity is further facilitated by p67(phox) or Noxa1, whereas the activators cancel the Noxo1-induced enhancement. On the other hand, the small GTPase Rac, essential for the gp91(phox) activity, is likely dispensable to the Nox3 system. Thus Nox3 functions together with p22(phox) as an enzyme constitutively producing superoxide, which can be distinctly regulated by combinatorial use of the organizers and activators. PMID- 15824104 TI - Cdc42 and Ras cooperate to mediate cellular transformation by intersectin-L. AB - Cdc42, a Ras-related GTP-binding protein, has been implicated in the regulation of the actin cytoskeleton, membrane trafficking, cell-cycle progression, and malignant transformation. We have shown previously that a Cdc42 mutant (Cdc42(F28L)), capable of spontaneously exchanging GDP for GTP (referred to as "fast-cycling"), transformed NIH 3T3 cells because of its ability to interfere with epidermal growth factor receptor (EGFR)-Cbl interactions and EGFR down regulation. To further examine the link between the hyperactivation of Cdc42 and its ability to alter EGFR signaling and thereby cause cellular transformation, we examined the effects of expressing different forms of the Cdc42-specific guanine nucleotide exchange factor, intersectin-L, in fibroblasts. Full-length intersectin-L exhibited little ability to stimulate nucleotide exchange on Cdc42, whereas a truncated version that contained five Src homology 3 (SH3) domains, the Dbl and pleckstrin homology domains (DH and PH domains, respectively), and a C2 domain (designated as SH3A-C2) showed modest guanine nucleotide exchange factor activity, whereas a form containing just the DH, PH, and C2 domains (DH-C2) strongly activated Cdc42. However, DH-C2 showed little ability to stimulate growth in low serum or colony formation in soft agar, whereas SH3A-C2 gave rise to a much stronger stimulation of cell growth in low serum and was highly effective in stimulating colony formation. Moreover, although SH3A-C2 strongly transformed fibroblasts, it differed from the actions of the Cdc42(F28L) mutant, as SH3A-C2 showed little ability to alter EGFR levels or the lifetime of EGF coupled signaling through ERK. Rather, we found that SH3A-C2 exhibited strong transforming activity through its ability to mediate cooperation between Ras and Cdc42. PMID- 15824105 TI - G protein beta2 subunit-derived peptides for inhibition and induction of G protein pathways. Examination of voltage-gated Ca2+ and G protein inwardly rectifying K+ channels. AB - Voltage-gated Ca2+ channels of the N-, P/Q-, and R-type and G protein inwardly rectifying K+ channels (GIRK) are modulated via direct binding of G proteins. The modulation is mediated by G protein betagamma subunits. By using electrophysiological recordings and fluorescence resonance energy transfer, we characterized the modulatory domains of the G protein beta subunit on the recombinant P/Q-type channel and GIRK channel expressed in HEK293 cells and on native non-L-type Ca2+ currents of cultured hippocampal neurons. We found that Gbeta2 subunit-derived deletion constructs and synthesized peptides can either induce or inhibit G protein modulation of the examined ion channels. In particular, the 25-amino acid peptide derived from the Gbeta2 N terminus inhibits G protein modulation, whereas a 35-amino acid peptide derived from the Gbeta2 C terminus induced modulation of voltage-gated Ca2+ channels and GIRK channels. Fluorescence resonance energy transfer (FRET) analysis of the live action of these peptides revealed that the 25-amino acid peptide diminished the FRET signal between G protein beta2gamma3 subunits, indicating a reorientation between G protein beta2gamma3 subunits in the presence of the peptide. In contrast, the 35 amino acid peptide increased the FRET signal between GIRK1,2 channel subunits, similarly to the Gbetagamma-mediated FRET increase observed for this GIRK subunit combination. Circular dichroism spectra of the synthesized peptides suggest that the 25-amino acid peptide is structured. These results indicate that individual G protein beta subunit domains can act as independent, separate modulatory domains to either induce or inhibit G protein modulation for several effector proteins. PMID- 15824106 TI - Novel role for cyclin-dependent kinase 2 in neuregulin-induced acetylcholine receptor epsilon subunit expression in differentiated myotubes. AB - Cyclin-dependent kinases (CDKs) are a family of evolutionarily conserved serine/threonine kinases. CDK2 acts as a checkpoint for the G(1)/S transition in the cell cycle. Despite a down-regulation of CDK2 activity in postmitotic cells, many cell types, including muscle cells, maintain abundant levels of CDK2 protein. This led us to hypothesize that CDK2 may have a function in postmitotic cells. We show here for the first time that CDK2 can be activated by neuregulin (NRG) in differentiated C2C12 myotubes. In addition, this activity is required for expression of the acetylcholine receptor (AChR) epsilon subunit. The switch from the fetal AChRgamma subunit to the adult-type AChRepsilon is required for synapse maturation and the neuromuscular junction. Inhibition of CDK2 activity with either the specific CDK2 inhibitory peptide Tat-LFG or by RNA interference abolished neuregulin-induced AChRepsilon expression. Neuregulin-induced activation of CDK2 also depended on the ErbB receptor, MAPK, and PI3K, all of which have previously been shown to be required for AChRepsilon expression. Neuregulin regulated CDK2 activity through coordinating phosphorylation of CDK2 on Thr-160, accumulation of CDK2 in the nucleus, and down-regulation of the CDK2 inhibitory protein p27 in the nucleus. In addition, we also observed a novel mechanism of regulation of CDK2 activity by a low molecular weight variant of cyclin E in response to NRG. These findings establish CDK2 as an intermediate molecule that integrates NRG-activated signals from both the MAPK and PI3K pathways to AChRepsilon expression and reveal an undiscovered physiological role for CDK2 in postmitotic cells. PMID- 15824107 TI - Specific interference of urokinase-type plasminogen activator receptor and matrix metalloproteinase-9 gene expression induced by double-stranded RNA results in decreased invasion, tumor growth, and angiogenesis in gliomas. AB - We have previously demonstrated the effectiveness of adenovirus-mediated expression of antisense urokinase-type plasminogen activator receptor (uPAR) and matrix metalloproteinase-9 (MMP-9) in inhibiting tumor invasion in vitro and ex vivo. However, the therapeutic effect of the adenovirus-mediated antisense approach was shown to be transient and required potentially toxic, high viral doses. In contrast, RNA interference (RNAi)-mediated gene targeting may be superior to the traditional antisense approach, because the target mRNA is completely degraded and the molar ratio of siRNA required to degrade the target mRNA is very low. Here, we have examined the siRNA-mediated target RNA degradation of uPAR and MMP-9 in human glioma cell lines. Using RNAi directed toward uPAR and MMP-9, we achieved specific inhibition of uPAR and MMP-9. This bicistronic construct (pUM) inhibited the formation of capillary-like structures in both in vitro and in vivo models of angiogenesis. We demonstrated that blocking the expression of these genes results in significant inhibition of glioma tumor invasion in Matrigel and spheroid invasion assay models. RNAi for uPAR and MMP-9 inhibited cell proliferation, and significantly reduced the levels of phosphorylated forms of MAPK, ERK, and AKT signaling pathway molecules when compared with parental and empty vector/scrambled vector-transfected SNB19 cells. Furthermore, using RNAi to simultaneously target two proteases resulted in total regression of pre-established intracerebral tumor growth. Our results provide evidence that the use of hairpin siRNA expression vectors for uPAR and MMP-9 may provide an effective tool for cancer therapy. PMID- 15824108 TI - Hydralysins, a new category of beta-pore-forming toxins in cnidaria. AB - Cnidaria are venomous animals that produce diverse protein and polypeptide toxins, stored and delivered into the prey through the stinging cells, the nematocytes. These include pore-forming cytolytic toxins such as well studied actinoporins. In this work, we have shown that the non-nematocystic paralytic toxins, hydralysins, from the green hydra Chlorohydra viridissima comprise a highly diverse group of beta-pore-forming proteins, distinct from other cnidarian toxins but similar in activity and structure to bacterial and fungal toxins. Functional characterization of hydralysins reveals that as soluble monomers they are rich in beta-structure, as revealed by far UV circular dichroism and computational analysis. Hydralysins bind erythrocyte membranes and form discrete pores with an internal diameter of approximately 1.2 nm. The cytolytic effect of hydralysin is cell type-selective, suggesting a specific receptor that is not a phospholipid or carbohydrate. Multiple sequence alignment reveals that hydralysins share a set of conserved sequence motifs with known pore-forming toxins such as aerolysin, epsilon-toxin, alpha-toxin, and LSL and that these sequence motifs are found in and around the poreforming domains of the toxins. The importance of these sequence motifs is revealed by the cloning, expression, and mutagenesis of three hydralysin isoforms that strongly differ in their hemolytic and paralytic activities. The correlation between the paralytic and cytolytic activities of hydralysin suggests that both are a consequence of receptor-mediated pore formation. Hydralysins and their homologues exemplify the wide distribution of beta-pore formers in biology and provide a useful model for the study of their molecular mode of action. PMID- 15824109 TI - Tumor suppression by the von Hippel-Lindau protein requires phosphorylation of the acidic domain. AB - The tumor suppressor function of the von Hippel-Lindau protein (pVHL) has previously been linked to its role in regulating hypoxia-inducible factor levels. However, VHL gene mutations suggest a hypoxia-inducible factor-independent function for the N-terminal acidic domain in tumor suppression. Here, we report that phosphorylation of the N-terminal acidic domain of pVHL by casein kinase-2 is essential for its tumor suppressor function. This post-translational modification did not affect the levels of hypoxia-inducible factor; however, it did change the binding of pVHL to another known binding partner, fibronectin. Cells expressing phospho-defective mutants caused improper fibronectin matrix deposition and demonstrated retarded tumor formation in mice. We propose that phosphorylation of the acidic domain plays a role in the regulation of proper fibronectin matrix deposition and that this may be relevant for the development of VHL-associated malignancies. PMID- 15824110 TI - Hypoxic augmentation of Ca2+ channel currents requires a functional electron transport chain. AB - The incidence of Alzheimer disease is increased following ischemic episodes, and we previously demonstrated that following chronic hypoxia (CH), amyloid beta (Abeta) peptide-mediated increases in voltage-gated L-type Ca(2+) channel activity contribute to the Ca(2+) dyshomeostasis seen in Alzheimer disease. Because in certain cell types mitochondria are responsible for detecting altered O(2) levels we examined the role of mitochondrial oxidant production in the regulation of recombinant Ca(2+) channel alpha(1C) subunits during CH and exposure to Abeta-(1-40). In wild-type (rho(+)) HEK 293 cells expressing recombinant L-type alpha(1C) subunits, Ca(2+) currents were enhanced by prolonged (24 h) exposure to either CH (6% O(2)) or Abeta-(1-40) (50 nm). By contrast the response to CH was absent in rho(0) cells in which the mitochondrial electron transport chain (ETC) was depleted following long term treatment with ethidium bromide or in rho(+) cells cultured in the presence of 1 microm rotenone. CH was mimicked in rho(0) cells by the exogenous production of O2(-.). by xanthine/xanthine oxidase. Furthermore Abeta-(1-40) enhanced currents in rho(0) cells to a degree similar to that seen in cells with an intact ETC. The antioxidants ascorbate (200 microm) and Trolox (500 microm) ablated the effect of CH in rho(+) cells but were without effect on Abeta-(1-40)-mediated augmentation of Ca(2+) current in rho(0) cells. Thus oxidant production in the mitochondrial ETC is a critical factor, acting upstream of amyloid beta peptide production in the up-regulation of Ca(2+) channels in response to CH. PMID- 15824111 TI - Alternative splicing switches the divalent cation selectivity of TRPM3 channels. AB - TRPM3 is a poorly understood member of the large family of transient receptor potential (TRP) ion channels. Here we describe five novel splice variants of TRPM3, TRPM3alpha1-5. These variants are characterized by a previously unknown amino terminus of 61 residues. The differences between the five variants arise through splice events at three different sites. One of these splice sites might be located in the pore region of the channel as indicated by sequence alignment with other, better-characterized TRP channels. We selected two splice variants, TRPM3alpha1 and TRPM3alpha2, that differ only in this presumed pore region and analyzed their biophysical characteristics after heterologous expression in human embryonic kidney 293 cells. TRPM3alpha1 as well as TRPM3alpha2 induced a novel, outwardly rectifying cationic conductance that was tightly regulated by intracellular Mg(2+). However, these two variants are highly different in their ionic selectivity. Whereas TRPM3alpha1-encoded channels are poorly permeable for divalent cations, TRPM3alpha2-encoded channels are well permeated by Ca(2+) and Mg(2+). Additionally, we found that currents through TRPM3alpha2 are blocked by extracellular monovalent cations, whereas currents through TRPM3alpha1 are not. These differences unambiguously show that TRPM3 proteins constitute a pore forming channel subunit and localize the position of the ion-conducting pore within the TRPM3 protein. Although the ionic selectivity of ion channels has traditionally been regarded as rather constant for a given channel-encoding gene, our results show that alternative splicing can be a mechanism to produce channels with very different selectivity profiles. PMID- 15824112 TI - Oxidation of a eukaryotic 2-Cys peroxiredoxin is a molecular switch controlling the transcriptional response to increasing levels of hydrogen peroxide. AB - Although activation of the AP-1-like transcription factor Pap1 in Schizosaccharomyces pombe is important for oxidative stress-induced gene expression, this activation is delayed at higher concentrations of peroxide. Here, we reveal that the 2-Cys peroxiredoxin (2-Cys Prx) Tpx1 is required for the peroxide-induced activation of Pap1. Tpx1, like other eukaryotic 2-Cys Prxs, is highly sensitive to oxidation, which inactivates its thioredoxin peroxidase activity. Our data suggest that the reduced thioredoxin peroxidase-active form of Tpx1 is required for the peroxide-induced oxidation and nuclear accumulation of Pap1. Indeed, in contrast to the previously described role for Tpx1 in the activation of the Sty1 stress-activated protein kinase by peroxide, we find that both catalytic cysteines of Tpx1 are required for Pap1 activation. Moreover, overexpression of the conserved sulfiredoxin Srx1, which interacts with and reduces Tpx1, allows rapid activation of Pap1 at higher concentrations of H(2)O(2). Conversely, loss of Srx1 prevents the reduction of oxidized Tpx1 and prolongs the inhibition of Pap1 activation. Collectively, these data suggest that redox regulation of the thioredoxin peroxidase activity of Tpx1 acts as a molecular switch controlling the transcriptional response to H(2)O(2). Furthermore, they reveal that a single eukaryotic 2-Cys Prx regulates peroxide signaling by multiple independent mechanisms. PMID- 15824113 TI - The T loop structure is dispensable for substrate recognition by tRNase ZL. AB - tRNA 3'-processing endoribonucleases (tRNase Z, or 3'-tRNase; EC 3.1.26.11) are enzymes that remove 3'-trailers from pre-tRNAs. An about 12-base-pair stem, a T loop-like structure, and a 3'-trailer were considered to be the minimum requirements for recognition by the long form (tRNase ZL) of tRNase Z; tRNase ZL can recognize and cleave a micro-pre-tRNA or a hooker/target RNA complex that resembles a micro-pre-tRNA. We examined four hook RNAs containing systematically weakened T stems for directing target RNA cleavage by tRNase ZL. As expected, the cleavage efficiency decreased with the decrease in T stem stability, and to our surprise, even the hook RNA that forms no T stem-loop-directed slight cleavage of the target RNA, suggesting that the T stem-loop structure is important but dispensable for substrate recognition by tRNase ZL. To analyze the effect of the T loop on substrate recognition, we compared the cleavage reaction for a micro pre-tRNA with that for a 12-base-pair double-stranded RNA, which is the same as the micro-pre-tRNA except for the lack of the T loop structure. The observed rate constant value for the double-stranded RNA was comparable with that for the micro pre-tRNA, whereas the K(d) value for the complex with the double-stranded RNA was much higher than that for the complex with the micro-pre-tRNA. These results suggest that the T loop structure is not indispensable for the recognition, although the interaction between the T loop and the enzyme exists. Cleavage assays for such double-stranded RNA substrates of various lengths suggested that tRNase ZL can recognize and cleave double-stranded RNA substrates that are longer than 5 base pairs and shorter than 20 base pairs. We also showed that double stranded RNA is not a substrate for the short form of tRNase Z. PMID- 15824114 TI - Laminin gamma3 chain binds to nidogen and is located in murine basement membranes. AB - Recently a novel laminin gamma3 chain was identified in mouse and human and shown to have the same modular structure as the laminin gamma1 chain. We expressed two fragments of the gamma3 chain in mammalian cells recombinantly. The first, domain VI/V, consisting of laminin N-terminal (domain VI) and four laminin-type epidermal growth factor-like (domain V) and laminin N-terminal modules, was shown to be essential for self-assembly of laminins. The other was domain III3-5, which consists of three laminin-type epidermal growth factor-like modules and is predicted to bind to nidogens. The gamma3 VI/V fragment was a poor inhibitor for laminin-1 polymerization as was the beta2 VI/V fragment. The gamma3 III3-5 fragment bound to nidogen-1 and nidogen-2 with lower affinity than the gamma1 III3-5 fragment. These data suggested that laminins containing the gamma3 chain may assemble networks independent of other laminins. Polyclonal antibodies raised against gamma3 VI/V and gamma3 III3-5 showed no cross-reaction with homologous fragments from the gamma1 and gamma2 chains of laminin and allowed the establishment of gamma chain-specific radioimmunoassays and light and electron microscopic immunostaining of tissues. This demonstrated a 20-100-fold lower content of the gamma3 chain compared with the gamma1 chain in various tissue extracts of adult mice. The expression of gamma3 chain was highly tissue specific. In contrast to earlier assumptions, the antibodies against the gamma3 chain showed light microscopic staining exclusively in basement membrane zones of adult and embryonic tissues, such as the brain, kidney, skin, muscle, and testis. Ultrastructural immunogold staining localized the gamma3 chain to basement membranes of these tissues. PMID- 15824115 TI - Crystal structures of the ferrous dioxygen complex of wild-type cytochrome P450eryF and its mutants, A245S and A245T: investigation of the proton transfer system in P450eryF. AB - Cytochrome P450eryF (CYP107A) from Saccaropolyspora ertherea catalyzes the hydroxylation of 6-deoxyerythronolide B, one of the early steps in the biosynthesis of erythromycin. P450eryF has an alanine rather than the conserved threonine that participates in the activation of dioxygen (O(2)) in most other P450s. The initial structure of P450eryF (Cupp-Vickery, J. R., Han, O., Hutchinson, C. R., and Poulos, T. L. (1996) Nat. Struct. Biol. 3, 632-637) suggests that the substrate 5-OH replaces the missing threonine OH group and holds a key active site water molecule in position to donate protons to the iron linked dioxygen, a critical step for the monooxygenase reaction. To probe the proton delivery system in P450eryF, we have solved crystal structures of ferrous wild-type and mutant (Fe(2+)) dioxygen-bound complexes. The catalytic water molecule that was postulated to provide protons to dioxygen is absent, although the substrate 5-OH group donates a hydrogen bond to the iron-linked dioxygen. The hydrogen bond network observed in the wild-type ferrous dioxygen complex, water 63-Glu(360)-Ser(246)-water 53-Ala(241) carbonyl in the I-helix cleft, is proposed as the proton transfer pathway. Consistent with this view, the hydrogen bond network in the O(2).A245S and O(2) .A245T mutants, which have decreased or no enzyme activity, was perturbed or disrupted, respectively. The mutant Thr(245) side chain also perturbs the hydrogen bond between the substrate 5-OH and dioxygen ligand. Contrary to the previously proposed mechanism, these results support the direct involvement of the substrate in O(2) activation but raise questions on the role water plays as a direct proton donor to the iron-linked dioxygen. PMID- 15824118 TI - PsaK2 subunit in photosystem I is involved in state transition under high light condition in the cyanobacterium Synechocystis sp. PCC 6803. AB - To avoid the photodamage, cyanobacteria regulate the distribution of light energy absorbed by phycobilisome antenna either to photosystem II or to photosystem I (PSI) upon high light acclimation by the process so-called state transition. We found that an alternative PSI subunit, PsaK2 (sll0629 gene product), is involved in this process in the cyanobacterium Synechocystis sp. PCC 6803. An examination of the subunit composition of the purified PSI reaction center complexes revealed that PsaK2 subunit was absent in the PSI complexes under low light condition, but was incorporated into the complexes during acclimation to high light. The growth of the psaK2 mutant on solid medium was inhibited under high light condition. We determined the photosynthetic characteristics of the wild type strain and the two mutants, the psaK1 (ssr0390) mutant and the psaK2 mutant, using pulse amplitude modulation fluorometer. Non-photochemical quenching, which reflects the energy transfer from phycobilisome to PSI in cyanobacteria, was higher in high light grown cells than in low light grown cells, both in the wild type and the psaK1 mutant. However, this change of non-photochemical quenching during acclimation to high light was not observed in the psaK2 mutant. Thus, PsaK2 subunit is involved in the energy transfer from phycobilisome to PSI under high light condition. The role of PsaK2 in state transition under high light condition was also confirmed by chlorophyll fluorescence emission spectra determined at 77 K. The results suggest that PsaK2-dependent state transition is essential for the growth of this cyanobacterium under high light condition. PMID- 15824116 TI - Enhanced proliferation of cultured human vascular smooth muscle cells linked to increased function of RNA-binding protein HuR. AB - In dividing cells, the RNA-binding protein HuR associates with and stabilizes labile mRNAs encoding proliferative proteins, events that are linked to the increased cytoplasmic presence of HuR. Here, assessment of HuR levels in various vascular pathologies (intimal hyperplasia, atherosclerosis and neointimal proliferation, sclerosis of arterialized saphenous venous graft, and fibromuscular dysplasia) revealed a distinct increase in HuR expression and cytoplasmic abundance within the intima and neointima layers. On the basis of these observations, we postulated a role for HuR in promoting the proliferation of vascular smooth muscle cells. To test this hypothesis directly, we investigated the expression, subcellular localization, and proliferative influence of HuR in human vascular smooth muscle cells (hVSMCs). Treatment of hVSMCs with platelet-derived growth factor increased HuR levels in the cytoplasm, thereby influencing the expression of metabolic, proliferative, and structural genes. Importantly, knockdown of HuR expression by using RNA interference caused a reduction of hVSMC proliferation, both basally and following platelet-derived growth factor treatment. We propose that HuR contributes to regulating hVSMC growth and homeostasis in pathologies associated with vascular smooth muscle proliferation. PMID- 15824117 TI - Overexpression of superoxide dismutase or glutathione peroxidase protects against the paraquat + maneb-induced Parkinson disease phenotype. AB - Oxidative stress has been implicated in the pathogenesis of Parkinson disease based on its role in the cascade of biochemical changes that lead to dopaminergic neuronal death. This study analyzed the role of oxidative stress as a mechanism of the dopaminergic neurotoxicity produced by the combined paraquat and maneb model of the Parkinson disease phenotype. Transgenic mice overexpressing either Cu,Zn superoxide dismutase or intracellular glutathione peroxidase and non transgenic mice were exposed to saline, paraquat, or the combination of paraquat + maneb twice a week for 9 weeks. Non-transgenic mice chronically exposed to paraquat + maneb exhibited significant reductions in locomotor activity, levels of striatal dopamine and metabolites, and dopaminergic neurons in the substantia nigra pars compacta. In contrast, no corresponding effects were observed in either Cu,Zn superoxide dismutase or glutathione peroxidase transgenic mice. Similarly, the increase in levels of lipid hydroperoxides in the midbrain and striatum of paraquat + maneb-treated non-transgenic mice was not detected in either Cu,Zn superoxide dismutase or glutathione peroxidase transgenic mice. To begin to determine critical pathways of paraquat + maneb neurotoxicity, the functions of cell death-inducing and protective mechanisms were analyzed. Even a single injection of paraquat + maneb in the non-transgenic treated group modulated several key pro- and anti-apoptotic proteins, including Bax, Bad, Bcl xL, and upstream stress-induced cascade. Collectively, these findings support the assertion that protective mechanisms against paraquat + maneb-induced neurodegeneration could involve modulation of the level of reactive oxygen species and alterations of the functions of specific signaling cascades. PMID- 15824119 TI - Isolation and characterization of novel cyclotides from Viola hederaceae: solution structure and anti-HIV activity of vhl-1, a leaf-specific expressed cyclotide. AB - Based on a newly established sequencing strategy featured by its efficiency, simplicity, and easy manipulation, the sequences of four novel cyclotides (macrocyclic knotted proteins) isolated from an Australian plant Viola hederaceae were determined. The three-dimensional solution structure of V. hederaceae leaf cyclotide-1 (vhl-1), a leaf-specific expressed 31-residue cyclotide, has been determined using two-dimensional (1)H NMR spectroscopy. vhl-1 adopts a compact and well defined structure including a distorted triple-stranded beta-sheet, a short 3(10) helical segment and several turns. It is stabilized by three disulfide bonds, which, together with backbone segments, form a cyclic cystine knot motif. The three-disulfide bonds are almost completely buried into the protein core, and the six cysteines contribute only 3.8% to the molecular surface. A pH titration experiment revealed that the folding of vhl-1 shows little pH dependence and allowed the pK(a) of 3.0 for Glu(3) and approximately 5.0 for Glu(14) to be determined. Met(7) was found to be oxidized in the native form, consistent with the fact that its side chain protrudes into the solvent, occupying 7.5% of the molecular surface. vhl-1 shows anti-HIV activity with an EC(50) value of 0.87 microm. PMID- 15824120 TI - SUMOylation of the polyglutamine repeat protein, ataxin-1, is dependent on a functional nuclear localization signal. AB - SUMO (small ubiquitin-like modifier) is a member of the ubiquitin family of proteins. SUMO targets include proteins involved in numerous roles including nuclear transport and transcriptional regulation. The previous finding that mutant ataxin-1[82Q] disrupted promyelocytic leukemia (PML) oncogenic domains prompted us to determine whether ataxin-1 disrupts another component of PML oncogenic domains, Sp100 (100-kDa Speckled protein). Similar to the PML protein, mutant ataxin-1[82Q] redistributed Sp100 to mutant ataxin-1[82Q] nuclear inclusions. Based on the ability of PML and Sp100 to be covalently modified by SUMO, we investigated the ability of ataxin-1 to be SUMOylated. SUMO-1 was found to covalently modify the polyglutamine repeat protein ataxin-1. There was a decrease in ataxin-1 SUMOylation in the presence of the expanded polyglutamine tract, ataxin-1[82Q]. The phospho-mutant, ataxin-1[82Q]-S776A, restored SUMO levels to those of wild-type ataxin-1[30Q]. SUMOylation of ataxin-1 was dependent on a functional nuclear localization signal. Ataxin-1 SUMOylation was mapped to at least five lysine residues. Lys(16), Lys(194) preceding the polyglutamine tract, Lys(610)/Lys(697) in the C-terminal ataxin high mobility group domain, and Lys(746) all contribute to ataxin-1 SUMOylation. PMID- 15824121 TI - Vitamin D receptor-dependent regulation of colon multidrug resistance-associated protein 3 gene expression by bile acids. AB - The multidrug resistance-associated protein 3 (MRP3) is a multispecific anion transporter that is capable of transporting a number of conjugated and unconjugated bile acids. Expression of the MRP3 gene is increased during pathological states associated with elevated bile acid concentrations indicating a role for this transporter in adaptive and homeostatic bile acid metabolism. Analysis of Mrp3 mRNA levels in various mouse tissues with known relevance and/or exposure to bile acids revealed the highest levels of basal expression in the colon followed in order by the liver, duodenum, jejunum, ileum, and kidney. Functional analysis of a murine Mrp3 promoter reporter construct revealed vitamin D receptor (VDR)-dependent activation by 1,25-dihydroxyvitamin D(3) (VD3), 9-cis retinoic acid (RA), and the cholestatic secondary bile acid, lithocholic acid (LCA). Using a series of deletion constructs combined with sequence analysis, a candidate VDR response element (VDRE) was identified between -1028 and -1014 bp of the Mrp3 promoter. Activation of the Mrp3 promoter in response to VD3, RA, or LCA, as well as binding of VDR/RXR heterodimers, was attenuated substantially by mutation of this VDRE. Treatment of mice with VD3 or LCA demonstrated in vivo modulation of the Mrp3 gene in colon but not in the liver. Reduction of endogenous VDR expression in colon adenocarcinoma MCA-38 cells by siRNA transfection was associated with reduced constitutive and inducible expression of the Mrp3 gene. These data support a regulatory role for the VDR in the protection of colon cells from bile acid toxicity through regulation of the Mrp3 expression. PMID- 15824122 TI - Sir2 and the acetyltransferase, Pat, regulate the archaeal chromatin protein, Alba. AB - The DNA binding affinity of Alba, a chromatin protein of the archaeon Sulfolobus solfataricus P2, is regulated by acetylation of lysine 16. Here we identify an acetyltransferase that specifically acetylates Alba on this residue. The effect of acetylation is to lower the affinity of Alba for DNA. Remarkably, the acetyltransferase is conserved not only in archaea but also in bacteria where it appears to play a role in metabolic regulation. Therefore, our data suggest that S. solfataricus has co-opted this bacterial regulatory system to generate a rudimentary form of chromatin regulation. PMID- 15824123 TI - Mycobacterium tuberculosis strains possess functional cellulases. AB - The genomes of various Mycobacterium tuberculosis strains encode proteins that do not appear to play a role in the growth or survival of the bacterium in its mammalian host, including some implicated in plant cell wall breakdown. Here we show that M. tuberculosis H37Rv does indeed possess a functional cellulase. The x ray crystal structure of this enzyme, in ligand complex forms, from 1.9 to 1.1A resolution, reveals a highly conserved substrate-binding cleft, which affords similar, and unusual, distortion of the substrate at the catalytic center. The endoglucanase activity, together with the existence of a putative membrane associated crystalline polysaccharide-binding protein, may reflect the ancestral soil origin of the Mycobacterium or hint at a previously unconsidered environmental niche. PMID- 15824124 TI - PTEN, but not SHIP2, suppresses insulin signaling through the phosphatidylinositol 3-kinase/Akt pathway in 3T3-L1 adipocytes. AB - Glucose homeostasis is controlled by insulin in part through the stimulation of glucose transport in muscle and fat cells. This insulin signaling pathway requires phosphatidylinositol (PI) 3-kinase-mediated 3'-polyphosphoinositide generation and activation of Akt/protein kinase B. Previous experiments using dominant negative constructs and gene ablation in mice suggested that two phosphoinositide phosphatases, SH2 domain-containing inositol 5'-phosphatase 2 (SHIP2) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) negatively regulate this insulin signaling pathway. Here we directly tested this hypothesis by selectively inhibiting the expression of SHIP2 or PTEN in intact cultured 3T3-L1 adipocytes through the use of short interfering RNA (siRNA). Attenuation of PTEN expression by RNAi markedly enhanced insulin-stimulated Akt and glycogen synthase kinase 3alpha (GSK-3alpha) phosphorylation, as well as deoxyglucose transport in 3T3-L1 adipocytes. In contrast, depletion of SHIP2 protein by about 90% surprisingly failed to modulate these insulin-regulated events under identical assay conditions. In control studies, no diminution of insulin signaling to the mitogen-activated protein kinases Erk1 and Erk2 was observed when either PTEN or SHIP2 were depleted. Taken together, these results demonstrate that endogenous PTEN functions as a suppressor of insulin signaling to glucose transport through the PI 3-kinase pathway in cultured 3T3-L1 adipocytes. PMID- 15824125 TI - Proteome dynamics during C2C12 myoblast differentiation. AB - Mouse-derived C2C12 myoblasts serve as an experimentally tractable model system for investigating the molecular basis of skeletal muscle cell specification and development. To examine the biochemical adaptations associated with myocyte formation comprehensively, we used large scale gel-free tandem mass spectrometry to monitor global proteome alterations throughout a time course analysis of the myogenic C2C12 differentiation program. The relative abundance of approximately 1,800 high confidence proteins was tracked across multiple time points using capillary scale multidimensional liquid chromatography coupled to high throughput shotgun sequencing. Hierarchical clustering of the resulting profiles revealed differential waves of expression of proteins linked to intracellular signaling, transcription, cytoarchitecture, adhesion, metabolism, and muscle contraction across the early, mid, and late stages of differentiation. Several hundred previously uncharacterized proteins were likewise detected in a stage-specific manner, suggesting novel roles in myogenesis and/or muscle function. These proteomic data are complementary to recent microarray-based studies of gene expression patterns in developing myotubes and provide a holistic framework for understanding how diverse biochemical processes are coordinated at the cellular level during skeletal muscle development. PMID- 15824127 TI - Optimal treatment of renal anaemia (OPTA): improving the efficacy and efficiency of renal anaemia therapy in haemodialysis patients receiving intravenous epoetin. AB - The medical care of renal anaemia has received much attention over the past decade, as nephrologists have recognized the increased therapeutic value of erythropoiesis-stimulating agents. The European Best Practice Guidelines and the US National Kidney Foundation's Kidney Disease Outcome Quality Initiative Guidelines have provided evidence-based advice on the optimal treatment of renal anaemia, and have recommended a target haemoglobin (Hb) level of 11 g/dl or 11-12 g/dl. Achieving this target Hb level has been shown to improve quality of life and reduce the rate of hospitalization; there is also good evidence to suggest that achieving adequate Hb levels reduces morbidity and mortality in patients with end-stage renal disease. In recent years, a number of factors have been identified that may counteract the positive action of epoetin therapy. These treatment-influencing factors include inadequate haemodialysis, absolute and functional iron deficiency, anticoagulant use, inflammation and infection. Each factor on its own may result in a substantial decrease in Hb levels, or an increase in epoetin requirements of up to 100%. Therefore, optimal and cost effective treatment can only be achieved by adequately managing all of the factors that potentially can influence anaemia in patients with chronic kidney disease. Large-scale, cross-sectional surveys, such as the European Survey on Anaemia Management and the Dialysis Outcomes and Practice Patterns Study, have shown that there is still room for improving the efficacy and efficiency of anaemia therapy. The Optimal Treatment of Renal Anaemia (OPTA) initiative aims to help both physicians and nurses improve renal anaemia management by "translating" the standards set in published guidelines into practical clinical advice. PMID- 15824128 TI - Results of the European Survey on Anaemia Management 2003 (ESAM 2003): current status of anaemia management in dialysis patients, factors affecting epoetin dosage and changes in anaemia management over the last 5 years. AB - The European Survey of Anaemia Management 2003 (ESAM 2003) was a 1 day randomized survey conducted to assess anaemia management in dialysis patients 4 years after the introduction of the European Best Practice Guidelines. The survey included 8100 patients from 11 European countries and Israel. Overall, haemoglobin (Hb) levels > or =11.0 g/dl, as recommended by the guidelines, were achieved in 66% of patients. Only 48% of patients had adequate iron status, with transferrin saturation values missing for 27% and functional or absolute iron deficiency reported for 17 and 9%, respectively. In order to identify factors affecting epoetin dose and Hb levels, the countries were divided into two groups based on the percentage of patients with Hb levels > or =11.0 g/dl (>70% in group 1 and 60 70% in group 2). The most probable causes for better management in group 1 were administration of higher epoetin doses and better monitoring and management of iron status. In patients with Hb <11.0 g/dl, mean epoetin-alpha/beta doses were significantly lower for subcutaneous than intravenous (i.v.) administration, whereas mean doses were similar for both routes in patients with Hb > or =11.0 g/dl. When standardized for Hb levels, the dose ratio of i.v. epoetin-alpha/beta to i.v. darbepoetin alfa was 176:1 (95% confidence interval, 166:1-189:1). Limited comparisons between the eight countries that participated in ESAM 2003 and the original ESAM revealed that many patients still have haemoglobin levels below the current recommendations despite significant improvements in management of renal anaemia over the last 5 years. PMID- 15824129 TI - Epoetin-associated pure red cell aplasia in patients with chronic kidney disease: solving the mystery. AB - A substantial increase in the incidence of pure red cell aplasia (PRCA) associated with recombinant human erythropoietin (epoetin) treatment occurred in 1998. The upsurge of antibody-mediated PRCA was almost exclusively associated with chronic kidney disease patients who received subcutaneous epoetin therapy and the formulation of epoetin-alpha distributed outside the USA (EPREX/ERYPO). A systematic programme of technical, immunological and epidemiological investigations was initiated to identify the possible causes. The potential causes were evaluated on the basis of the following criteria: temporal correlation with the increase in incidence of PRCA, significant difference between EPREX/ERYPO and other epoetin products, sufficient concentration in the product to elicit a weak immune response, evidence of immunogenic activity in animals supportive, and consistent with available clinical data. Organic compounds that were leached from rubber stoppers through the action of polysorbate 80 were detected in pre-filled syringes with uncoated rubber stoppers containing polysorbate 80-formulated EPREX/ERYPO (introduced outside the USA in 1998). The leachates were not present when the stoppers were coated, in the product formulated with human serum albumin or in other epoetin products. The adjuvant activity of the leachates was demonstrated in mice. The incidence of PRCA was significantly higher in patients exposed to the polysorbate 80 formulation of epoetin-alpha delivered from pre-filled syringes with uncoated rubber stoppers, which were recalled in 2003, than in patients exposed to the same formulation from syringes with coated rubber stoppers. In conclusion, these data strongly suggest that leachates were the critical contributory factor in the increased incidence of antibody-mediated PRCA attributed to EPREX/ERYPO. PMID- 15824130 TI - Proteomic and genomic characterization of chromatin complexes at a boundary. AB - We have dissected specialized assemblies on the Saccharomyces cerevisiae genome that help define and preserve the boundaries that separate silent and active chromatin. These assemblies contain characteristic stretches of DNA that flank particular regions of silent chromatin, as well as five distinctively modified histones and a set of protein complexes. The complexes consist of at least 15 chromatin-associated proteins, including DNA pol epsilon, the Isw2-Itc1 and Top2 chromatin remodeling proteins, the Sas3-Spt16 chromatin modifying complex, and Yta7, a bromodomain-containing AAA ATPase. We show that these complexes are important for the faithful maintenance of an established boundary, as disruption of the complexes results in specific, anomalous alterations of the silent and active epigenetic states. PMID- 15824131 TI - ZW10 links mitotic checkpoint signaling to the structural kinetochore. AB - The mitotic checkpoint ensures that chromosomes are divided equally between daughter cells and is a primary mechanism preventing the chromosome instability often seen in aneuploid human tumors. ZW10 and Rod play an essential role in this checkpoint. We show that in mitotic human cells ZW10 resides in a complex with Rod and Zwilch, whereas another ZW10 partner, Zwint-1, is part of a separate complex of structural kinetochore components including Mis12 and Ndc80-Hec1. Zwint-1 is critical for recruiting ZW10 to unattached kinetochores. Depletion from human cells or Xenopus egg extracts is used to demonstrate that the ZW10 complex is essential for stable binding of a Mad1-Mad2 complex to unattached kinetochores. Thus, ZW10 functions as a linker between the core structural elements of the outer kinetochore and components that catalyze generation of the mitotic checkpoint-derived "stop anaphase" inhibitor. PMID- 15824132 TI - Inhibition of the anaphase-promoting complex by the Xnf7 ubiquitin ligase. AB - Degradation of specific protein substrates by the anaphase-promoting complex/cyclosome (APC) is critical for mitotic exit. We have identified the protein Xenopus nuclear factor 7 (Xnf7) as a novel APC inhibitor able to regulate the timing of exit from mitosis. Immunodepletion of Xnf7 from Xenopus laevis egg extracts accelerated the degradation of APC substrates cyclin B1, cyclin B2, and securin upon release from cytostatic factor arrest, whereas excess Xnf7 inhibited APC activity. Interestingly, Xnf7 exhibited intrinsic ubiquitin ligase activity, and this activity was required for APC inhibition. Unlike other reported APC inhibitors, Xnf7 did not associate with Cdc20, but rather bound directly to core subunits of the APC. Furthermore, Xnf7 was required for spindle assembly checkpoint function in egg extracts. These data suggest that Xnf7 is an APC inhibitor able to link spindle status to the APC through direct association with APC core components. PMID- 15824133 TI - PKCeta is required for beta1gamma2/beta3gamma2- and PKD-mediated transport to the cell surface and the organization of the Golgi apparatus. AB - Protein kinase D (PKD) binds to a pool of diacylglycerol (DAG) in the TGN and undergoes a process of activation that involves heterotrimeric GTP-binding protein subunits betagamma to regulate membrane fission. This fission reaction is used to generate transport carriers at the TGN that are en route to the cell surface. We now report that PKD is activated specifically by G protein subunit beta1gamma2 and beta3gamma2 via the Golgi apparatus-associated PKCeta. Compromising the kinase activity of PKCeta-inhibited protein transport from TGN to the cell surface. Expression of constitutively activated PKCeta caused Golgi fragmentation, which was inhibited by a kinase inactive form of PKD. Our findings reveal that betagamma, PKCeta, and PKD act in series to generate transport carriers from the TGN and their overactivation results in complete vesiculation of the Golgi apparatus. PMID- 15824134 TI - The p38alpha/beta MAPK functions as a molecular switch to activate the quiescent satellite cell. AB - Somatic stem cells cycle slowly or remain quiescent until required for tissue repair and maintenance. Upon muscle injury, stem cells that lie between the muscle fiber and basal lamina (satellite cells) are activated, proliferate, and eventually differentiate to repair the damaged muscle. Satellite cells in healthy muscle are quiescent, do not express MyoD family transcription factors or cell cycle regulatory genes and are insulated from the surrounding environment. Here, we report that the p38alpha/beta family of mitogen-activated protein kinases (MAPKs) reversibly regulates the quiescent state of the skeletal muscle satellite cell. Inhibition of p38alpha/beta MAPKs (a) promotes exit from the cell cycle, (b) prevents differentiation, and (c) insulates the cell from most external stimuli allowing the satellite cell to maintain a quiescent state. Activation of satellite cells and p38alpha/beta MAPKs occurs concomitantly, providing further support that these MAPKs function as a molecular switch for satellite cell activation. PMID- 15824135 TI - An internal GAP domain negatively regulates presynaptic dynamin in vivo: a two step model for dynamin function. AB - The mechanism by which the self-assembling GTPase dynamin functions in vesicle formation remains controversial. Point mutations in shibire, the Drosophila dynamin, cause temperature-sensitive (ts) defects in endocytosis. We show that the ts2 mutation, which occurs in the switch 2 region of dynamin's GTPase domain, compromises GTP binding affinity. Three second-site suppressor mutations, one in the switch 1 region of the GTPase domain and two in the GTPase effector domain (GED), dynamin's putative GAP, fully rescue the shi(ts2) defects in synaptic vesicle recycling. The functional rescue in vivo correlates with a reduction in both the basal and assembly-stimulated GTPase activity in vitro. These findings demonstrate that GED is indeed an internal dynamin GAP and establish that, as for other GTPase superfamily members, dynamin's function in vivo is negatively regulated by its GAP activity. Based on these and other observations, we propose a two-step model for dynamin during vesicle formation in which an early regulatory GTPase-like function precedes late, assembly-dependent steps during which GTP hydrolysis is required for vesicle release. PMID- 15824136 TI - A Rho family GTPase controls actin dynamics and tip growth via two counteracting downstream pathways in pollen tubes. AB - Tip growth in neuronal cells, plant cells, and fungal hyphae is known to require tip-localized Rho GTPase, calcium, and filamentous actin (F-actin), but how they interact with each other is unclear. The pollen tube is an exciting model to study spatiotemporal regulation of tip growth and F-actin dynamics. An Arabidopsis thaliana Rho family GTPase, ROP1, controls pollen tube growth by regulating apical F-actin dynamics. This paper shows that ROP1 activates two counteracting pathways involving the direct targets of tip-localized ROP1: RIC3 and RIC4. RIC4 promotes F-actin assembly, whereas RIC3 activates Ca(2+) signaling that leads to F-actin disassembly. Overproduction or depletion of either RIC4 or RIC3 causes tip growth defects that are rescued by overproduction or depletion of RIC3 or RIC4, respectively. Thus, ROP1 controls actin dynamics and tip growth through a check and balance between the two pathways. The dual and antagonistic roles of this GTPase may provide a unifying mechanism by which Rho modulates various processes dependent on actin dynamics in eukaryotic cells. PMID- 15824138 TI - Retraction. PMID- 15824139 TI - A risk prediction tool for melanoma? PMID- 15824137 TI - Laminin-sulfatide binding initiates basement membrane assembly and enables receptor signaling in Schwann cells and fibroblasts. AB - Endoneurial laminins (Lms), beta1-integrins, and dystroglycan (DG) are important for Schwann cell (SC) ensheathment and myelination of axons. We now show that SC expression of galactosyl-sulfatide, a Lm-binding glycolipid, precedes that of Lms in developing nerves. This glycolipid anchors Lm-1 and -2 to SC surfaces by binding to their LG domains and enables basement membrane (BM) assembly. Revealingly, non-BM-forming fibroblasts become competent for BM assembly when sulfatides are intercalated into their cell surfaces. Assembly is characterized by coalescence of sulfatide, DG, and c-Src into a Lm-associated complex; by DG dependent recruitment of utrophin and Src activation; and by integrin-dependent focal adhesion kinase phosphorylation. Collectively, our findings suggest that sulfated glycolipids are key Lm anchors that determine which cell surfaces can assemble Lms to initiate BM assembly and DG- and integrin-mediated signaling. PMID- 15824140 TI - The biology of tobacco and nicotine: bench to bedside. AB - Strong epidemiologic evidence links smoking and cancer. An increased understanding of the molecular biology of tobacco-related cancers could advance progress toward improving smoking cessation and patient management. Knowledge gaps between tobacco addiction, tumorigenesis, and cancer brought an interdisciplinary group of investigators together to discuss "The Biology of Nicotine and Tobacco: Bench to Bedside." Presentations on the signaling pathways and pathogenesis in tobacco-related cancers, mouse models of addiction, imaging and regulation of nicotinic receptors, the genetic basis for tobacco carcinogenesis and development of lung cancer, and molecular mechanisms of carcinogenesis were heard. Importantly, new opportunities to use molecular biology to identify and abrogate tobacco-mediated carcinogenesis and to identify high-risk individuals were recognized. PMID- 15824141 TI - Epidemiology and prevention of breast cancer. PMID- 15824142 TI - Chlamydia pneumoniae and lung cancer: epidemiologic evidence. AB - Chlamydia pneumoniae is a common cause of acute respiratory infection and has been hypothesized to cause several chronic diseases, including lung cancer. The purpose of this article is to identify, describe, and critically examine the published studies on the association between C. pneumoniae infection and risk of lung cancer. In the six studies identified, previous C. pneumoniae infection was defined on the basis of serologic criteria, which varied between studies. All studies reported elevated relative risk estimates for the association of serologic evidence of infection and risk of lung cancer. The three studies in which past infection was defined based on testing of prediagnostic blood specimens tended to have weaker results (odds ratio range, 1.2-2.1) than those based on postdiagnostic blood specimens (odds ratio range, 1.4-9.9). Selection bias, measurement error, and inadequate control for confounding are concerns in some of these studies. Nevertheless, results were relatively consistent, supporting a causal association. Inflammation caused by chronic infection with C. pneumoniae may be involved in the carcinogenic process but this relationship will be difficult to further define through serologic data. To better understand the nature of this association, both experimental study designs, such as those based on animal models or randomized controlled antibiotic treatment trials in humans, and observational study designs (e.g., studies that involve detection of C. pneumoniae in pulmonary specimens obtained before cancer onset) could be explored and may shed additional light on this important association. PMID- 15824143 TI - Physical activity and risk of endometrial cancer: a report from the Shanghai endometrial cancer study. AB - We evaluated the type and amount of physical activity associated with risk of endometrial cancer. In this population-based case-control study, in-person interviews were completed among 832 incident endometrial cancer cases and 846 age matched controls. Physical activity from exercise, household activities, and transportation was assessed in adolescence and adulthood, as was lifetime occupational activity. Logistic regression was used to estimate adjusted odds ratios (OR) and 95% confidence limits (95% CL). Women reporting exercise participation in both adolescence and adulthood were at nearly a 40% reduced risk (OR, 0.63; 95% CL, 0.42-0.95), compared with women reporting no exercise in either life period. Postmenopausal women who initiated exercise in adulthood were also at reduced risk (OR, 0.76; 95% CL, 0.56-1.02). Reductions in risk were also observed for common lifestyle activities, including household activity (both life periods) and walking for transportation (adulthood). Examination of the independent and combined effect of exercise and lifestyle activities revealed that women with less active lifestyles but who reported exercise were at 35% reduced risk (OR, 0.65; 95% CL, 0.41-1.02), whereas nonexercisers with more active lifestyles were at 40% to 45% reduced risk. These findings suggest that both lifestyle activities of lower intensity (e.g., walking and doing household chores) and intentional exercise can reduce endometrial cancer risk. PMID- 15824144 TI - Ki-67 expression in benign breast ductal cells obtained by random periareolar fine needle aspiration. AB - Ki-67 expression in ductal cells obtained by random periareolar fine needle aspiration (RPFNA) is currently being used as a response biomarker in phase II breast cancer chemoprevention trials; however, Ki-67 in RPFNA has not been well studied as a risk predictor for cancer, which would support its use as a response indicator. We examined the expression of Ki-67 in RPFNA specimens with hyperplasia +/- atypia obtained from 147 women at high risk for development of breast cancer. Median Ki-67 was 1.4% (range 0-24%). Ki-67 was higher in specimens from women < 50 versus those > or = 50 (median 2% versus 0.6%; P = 0.006) and from premenopausal women versus postmenopausal women (P = 0.037); however, hormone replacement therapy (predominately low-dose estrogen without progestins) had no effect. By univariate analysis, Ki-67 was positively correlated with ductal cell number (P = 0.001) and hyperplasia with atypia (P = 0.007). By multivariable analysis, the proportion of ductal cells expressing Ki-67 was again predicted by cell number, which, in turn, was predicted by cytologic atypia. The association of Ki-67 expression with cytologic atypia, a known risk factor for development of breast cancer, provides preliminary justification for its use as a response biomarker in phase II chemoprevention trials. PMID- 15824145 TI - Retinoic acid receptor-beta2 promoter methylation in random periareolar fine needle aspiration. AB - Methylation of the retinoic acid receptor-beta2 (RARbeta2) P2 promoter is hypothesized to be an important mechanism for loss of RARbeta2 function during early mammary carcinogenesis. The frequency of RARbeta2 P2 methylation was tested in (a) 16 early stage breast cancers and (b) 67 random periareolar fine needle aspiration (RPFNA) samples obtained from 38 asymptomatic women who were at increased risk for breast cancer. Risk was defined as either (a) 5-year Gail risk calculation > or = 1.7%; (b) prior biopsy exhibiting atypical hyperplasia, lobular carcinoma in situ, or ductal carcinoma in situ; or (c) known BRCA1/2 mutation carrier. RARbeta2 P2 promoter methylation was assessed at two regions, M3 (-51 to 162 bp) and M4 (104-251 bp). In early stage cancers, M4 methylation was observed in 11 of 16 (69%) cases; in RPFNA samples, methylation was present at M3 and M4 in 28 of 56 (50%) and 19 of 56 (38%) cases, respectively. RPFNAs were stratified for cytologic atypia using the Masood cytology index. The distribution of RARbeta2 P2 promoter methylation was reported as a function of increased cytologic abnormality. Methylation at both M3 and M4 was observed in (a) 0 of 10 (0%) of RPFNAs with Masood scores of < or = 10 (nonproliferative), (b) 3 of 20 (15%) with Masood scores of 11 to 12 (low-grade proliferative), (c) 3 of 10 (30%) with Masood scores of 13 (high-grade proliferative), and (d) 7 of 14 (50%) with Masood scores of 14 of 15 (atypia). Results from this study indicate that the RARbeta2 P2 promoter is frequently methylated (69%) in primary breast cancers and shows a positive association with increasing cytologic abnormality in RPFNA. PMID- 15824146 TI - Postmenopausal breast cancer risk and cumulative number of menstrual cycles. AB - OBJECTIVE: To explore whether the lifetime cumulative number of menstrual cycles, as an index for total exposure to endogenous estrogens, and the number of menstrual cycles until a first full-term pregnancy (FFTP), are associated with breast cancer risk in postmenopausal women. METHODS: Population-based study with data from the Prospect-European Prospective Investigation into Cancer and Nutrition study. Naturally menopausal participants were eligible (n = 6,718). The cumulative number of menstrual cycles was computed in 6,031 (90%) women. We calculated the number of cycles until FFTP among parous participants. The number of menstrual cycles was impossible to compute in women who reported to be always irregular; therefore, we added the "always irregular" category in the analysis. During the 46,746 person-years of follow-up, 168 breast cancer cases were identified. Cox regression models were used and adjustments were made to account for potential confounders. RESULTS: Even when our data does not show a clear linear gradient, we observed an increased breast cancer risk in women with a higher number of cumulative menstrual cycles in their lifetime. Using < or = 415 cycles as reference, the hazard ratio for the irregular group, 416-453, 454-490, and > or = 491 cycles was 1.11 (.56, 2.19), 1.88 (1.14, 3.12), 1.74 (1.05, 2.87), and 1.80 (1.09, 2.96), respectively. Although not statistically significant, and of less magnitude, the risk estimates for the number of cycles before FFTP showed the same tendency. CONCLUSION: Among women who underwent natural menopause, a higher number of menstrual cycles in lifetime, reflecting a longer exposure to endogenous estrogens, is associated with an increased breast cancer risk. PMID- 15824147 TI - Flavonoids and breast cancer risk in Italy. AB - Few epidemiologic studies have investigated the potential relation between flavonoids and breast cancer risk. We have applied recently published data on the composition of foods and beverages in terms of six principal classes of flavonoids (i.e., flavanones, flavan-3-ols, flavonols, flavones, anthocyanidines, and isoflavones) on dietary information collected in a large-case control study of breast cancer conducted in Italy between 1991 and 1994. The study included 2,569 women with incident, histologically confirmed breast cancer, and 2,588 hospital controls. Odds ratios (OR) and 95% confidence intervals were estimated by multiple logistic regression models. After allowance for major confounding factors and energy intake, a reduced risk of breast cancer was found for increasing intake of flavones (OR, 0.81, for the highest versus the lowest quintile; P-trend, 0.02), and flavonols (OR, 0.80; P-trend, 0.06). No significant association was found for other flavonoids, including flavanones (OR, 0.95), flavan-3-ols (OR, 0.86), anthocyanidins (OR, 1.09), as well as for isoflavones (OR, 1.05). The findings of this large study of an inverse association between flavones and breast cancer risk confirm the results of a Greek study. PMID- 15824148 TI - Lack of association between EBV and breast carcinoma. AB - Multiple conflicting findings have been presented which indicate that EBV may be found in anywhere from 0% to 51% of breast carcinomas. When EBV has been found causally associated with other human cancers, its DNA and one or more of its viral products have been detected in most tumor cells of a given biopsy. To test whether EBV has such an association with breast cancer, we measured the number of viral DNA molecules per cell in matched normal and tumor biopsies from 45 patients using real-time quantitative PCR. In no case could EBV DNA consistently be detected, with either of two different probes, at levels above 0.1 molecules per cell in two sections of the tumor samples. These levels of detection match those detected in EBV-negative cell lines and therefore likely represent noise in the assays. Equally importantly, the distribution of these low signals was the same between tumors and their matched normal controls. We conclude that EBV does not contribute to the development of breast cancers as it does to epithelial cancers such as nasopharyngeal and gastric carcinomas or to Burkitt's and Hodgkin's lymphomas. PMID- 15824149 TI - Cyp17, urinary sex steroid levels and breast cancer risk in postmenopausal women. AB - Endogenous sex hormones play an important role in the etiology of breast cancer. Polymorphisms in genes encoding for enzymes involved in steroidogenesis may therefore play a role in breast cancer risk. Cytochrome P450c17alpha (Cyp17) functions at key branch points in human steroidogenesis. A T-->C transition (A1 and A2 allele) in the 5' untranslated region may be associated with increased expression of Cyp17. Using a case-cohort design, we studied the effects of the A2 allele on endogenous sex hormone levels and breast cancer risk within a large population-based cohort (n = 9,349) in the Netherlands (the DOM-cohort). Cyp17 genotype was determined in 335 incident postmenopausal breast cancer cases, which occurred after follow-up (median time to follow-up, 19 years) of the entire cohort, and in a random sample of 373 women (subcohort). Concentrations of estrone (E1), estradiol (E2), testosterone, 5alpha-androstane-3alpha, 17beta-diol (3alphaD), and creatinine were measured in first-morning urine samples. Only among women with body mass index (BMI) < 25 kg/m2 was the A2A2 genotype associated with higher levels of E1, E2, and 3alphaD compared with a group of women with either the A1A1 or the A1A2 genotype (e.g., geometric means of E(1) in ng/mg(creatinine): A2A2, 2.23; A1A1/A1A2, 1.47; P = 0.03). Adjusted breast cancer rate ratios for women with the A1A2 or A2A2 genotype compared with women with the A1A1 genotype were 0.96 (0.68-1.37) and 0.80 (0.47-1.35), respectively. These results did not differ between women with low and high BMI. In conclusion, this paper shows that women with low BMI and the A2A2 genotype had higher endogenous sex steroid levels compared with women with the A1A1 genotype. However, these increased sex steroid levels are not translated into an increased breast cancer risk in these women. PMID- 15824150 TI - Genetic polymorphisms of ataxia telangiectasia mutated and breast cancer risk. AB - To evaluate the role of genetic polymorphisms of ataxia telangiectasia mutated (ATM) in the etiology of breast cancer, a hospital-based case-control study was conducted in Korea. Nine-hundred ninety-six histologically confirmed incident breast cancer cases and 1,181 cancer-free controls were recruited in Seoul between 1995 and 2003. Genotypes of the ATM polymorphisms-5144A > T, IVS21 + 1049T > C, IVS33 - 55T > C, IVS34 + 60G > A, and 3393T > G were determined by the 5'-nuclease assay. Individual haplotypes were estimated from genotype data by a Bayesian method. Five ATM alleles were found to be in strong linkage disequilibrium (D' > 0.82; P < 0.001). Haplotype frequencies were significantly different between cases and controls (chi2 test, P < 0.001). The ATM IVS21 + 1049 TC or CC, IVS34 + 60 GA or AA, and 3393 TG or GG genotypes were associated with increased breast cancer risk, particularly in premenopausal women [odds ratios (OR), 1.51; 95% confidence interval (CI), 1.11-2.05; OR, 1.42; 95% CI, 1.08-1.88; and OR, 1.37; 95% CI, 1.04-1.80, respectively]. Compared with diploid of TCCAG:TCCAG, the most common haplotype, the ATTGT:ATTGT was associated with decreased risk of breast cancer with borderline significance (OR, 0.77; 95% CI, 0.58-1.04) and TCCAG:ATCGT and ATTGT:ACCAG were associated with increased breast cancer risk (OR, 2.30; 95% CI, 1.18-4.48 and OR, 2.43; 95% CI, 1.1.07-5.52, respectively) after adjusting for age, education, age at first full-term pregnancy, parity, family history of breast cancer, alcohol consumption, and smoking. As the number of ATTGT haplotype decreased, the risk of breast cancer increased (P for trend < 0.01). Our results thus suggest that genetic polymorphisms of ATM play an important role in the development of breast cancer in Korean women. PMID- 15824151 TI - Increased retinoic acid receptor-beta4 correlates in vivo with reduced retinoic acid receptor-beta2 in esophageal squamous cell carcinoma. AB - Different retinoic acid receptor-beta (RAR-beta) isoforms seem to have contrasting biological effects in human carcinogenesis. Both in vitro and in vivo data indicate that RAR-beta2 expression is frequently lost or reduced (and transfecting RAR-beta2 suppresses growth and promotes apoptosis) in various cancer cells and tissues, whereas RAR-beta4 expression is increased in several cancer cell lines. To clarify the effects of different RAR-beta isoforms in esophageal carcinogenesis, we used real-time quantitative reverse transcription PCR to assess in vivo RAR-beta mRNA levels in specimens of normal and malignant human esophageal tissue, comparing these levels with each other and the expressions of other genes. RAR-beta2 mRNA expression was significantly reduced (i.e., lower in cancer than normal tissue) in 67% (18 of 27, P = 0.001) and RAR beta(4) mRNA was increased in 52% (14 of 27, P = 0.054) of our esophageal cancer cases. The expressions of RAR-beta1, chicken ovalbumin upstream promoter transcription factor-I (COUP-TFI), COUP-TFII, and peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA were reduced, whereas epidermal growth factor receptor and cyclin D1 expressions were increased in tumor compared with in normal tissues. Reduced RAR-beta2 expression correlated with increased RAR beta4 expression (P = 0.002) and with the suppression of COUP-TFI and COUP-TFII (P = 0.050 and 0.023, respectively) in tumor samples. These are the first in vivo expression patterns of RAR-beta2 and RAR-beta4 reported in humans or animals and support the in vitro data on these isoforms and their contrasting biological effects in human carcinogenesis. PMID- 15824152 TI - Frequent methylation of eyes absent 4 gene in Barrett's esophagus and esophageal adenocarcinoma. AB - Most esophageal adenocarcinomas arise within Barrett's esophagus but the cause of this increasingly prevalent condition remains unknown. Early detection improves survival and discriminant screening markers for Barrett's esophagus and cancer are needed. This study was designed to explore the natural history of eyes absent 4 (EYA4) gene methylation in the neoplastic progression of Barrett's esophagus and to evaluate methylated EYA4 as a candidate marker. Aberrant promoter methylation of EYA4 was studied by methylation-specific PCR using bisulfite treated DNA from esophageal adenocarcinomas, Barrett's esophagus, and normal epithelia, and then confirmed by sequencing. Eight cancer cell lines were treated with the demethylation agent 5-aza-2'-deoxycytidine, and EYA4 mRNA expression with and without treatment was quantified by real-time reverse-transcription PCR. EYA4 hypermethylation was detected in 83% (33 of 40) of esophageal adenocarcinomas and 77% (27 of 35) of Barrett's tissues, but only in 3% (2 of 58) of normal esophageal and gastric mucosa samples (P < 0.001). The unmethylated cancer cell lines had much higher EYA4 mRNA expression than the methylated cancer cell lines. Demethylation caused by 5-aza-2'-deoxycytidine increased the mRNA expression level by a median of 3.2-fold in methylated cells, but its effect on unmethylated cells was negligible. Results indicate that aberrant promoter methylation of EYA4 is very common during tumorigenesis in Barrett's esophagus, occurs in early metaplasia, seems to be an important mechanism of down-regulating EYA4 expression, and represents an intriguing candidate marker for Barrett's metaplasia and esophageal cancer. PMID- 15824153 TI - Dietary marine n-3 fatty acids in relation to risk of distal colorectal adenoma in women. AB - Epidemiologic studies of dietary marine n-3 fatty acids and risk of colorectal cancer have been inconsistent, and their relation to risk of colorectal adenoma has not been evaluated in detail. We examined dietary marine n-3 fatty acids and the ratio of marine n-3 to total n-6 fatty acids (n-3/n-6 ratio) in relation to risk of adenoma of the distal colon or rectum among 34,451 U.S. women who were initially free of colorectal cancer or polyps, who completed a semiquantitative food frequency questionnaire in 1980, and who underwent endoscopy from 1980 to 1998. We documented 1,719 distal colorectal adenoma cases (705 large adenomas, 897 small adenomas, 1,280 distal colon adenomas, and 505 rectal adenomas) during 18 years of follow-up. Neither dietary marine n-3 fatty acids nor n-3/n-6 ratio were associated with risk of total distal colorectal adenoma after adjustment for age and established risk factors [multivariable relative risk (RR) for extreme quintiles of dietary marine n-3 fatty acids = 1.04; 95% confidence interval (95% CI), 0.84-1.27, P(trend) = 0.66; RR for extreme quintiles of n-3/n-6 ratio = 1.02; 95% CI, 0.83-1.25; P(trend) = 0.86]. Similarly, no significant associations were observed separately for distal colon or rectal adenoma. However, higher intake of dietary marine n-3 fatty acids was nonsignificantly but suggestively inversely associated with large adenoma (RR, 0.74; 95% CI, 0.54-1.01; P(trend) = 0.16) but directly associated with small adenoma (RR, 1.36; 95% CI, 1.02-1.81; P(trend) = 0.09). Our findings do not support the hypothesis that a higher intake of marine n-3 fatty acids or a higher n-3/n-6 ratio reduces the risk of distal colorectal adenoma but are suggestive that higher intake may reduce the progression of small adenomas to large adenomas. PMID- 15824154 TI - Fiber intake and incidence of colorectal cancer among 76,947 women and 47,279 men. AB - Prospective cohort studies have consistently found no important link between fiber intake and risk of colorectal cancer. The recent large, prospective European Prospective Investigation into Cancer and Nutrition has challenged this paradigm by suggesting significant protection by high fiber intake. We prospectively investigated the association of fiber intake with the incidence of colon and rectal cancers in two large cohorts: the Nurses' Health Study (76,947 women) and the Health Professionals Follow-up Study (47,279 men). Diet was assessed repeatedly in 1984, 1986, 1990, and 1994 among women and in 1986, 1990, and 1994 among men. The incidence of cancer of the colon and rectum was ascertained up to the year 2000. Relative risk estimates were calculated using a Cox proportional hazards model simultaneously controlling for potential confounding variables. During follow-up including 1.8 million person-years and 1,596 cases of colorectal cancer, we found little association with fiber intake after controlling for confounding variables. The hazard ratio for a 5-g/d increase in fiber intake was 0.91 (95% confidence interval, 0.87-0.95) after adjusting for covariates used in the European Prospective Investigation into Cancer and Nutrition study and 0.99 (95% confidence interval, 0.95-1.04) after adjusting for additional confounding variables. Our data from two large prospective cohorts with long follow-up and repeated assessment of fiber intake and of a large number of potential confounding variables do not indicate an important association between fiber intake and colorectal cancer but reveal considerable confounding by other dietary and lifestyle factors. PMID- 15824155 TI - A prospective study of C-peptide, insulin-like growth factor-I, insulin-like growth factor binding protein-1, and the risk of colorectal cancer in women. AB - Hyperinsulinemia, hyperglycemia, and elevated insulin-like growth factor (IGF)-1 levels have been implicated in the etiology of colorectal cancer. However, the joint effects of insulin and IGF-I have not been considered, and whether hyperinsulinemia or hyperglycemia is more etiologically relevant is unclear. IGF binding protein-1 (IGFBP-1) has been hypothesized to mediate the effects of insulin, but epidemiologic data on IGFBP-1 are sparse. We conducted a nested case control study among the 32,826 women of the Nurses' Health Study who provided a blood sample in 1989 to 1990. After excluding diabetics, we confirmed 182 incident colorectal cancer cases over 10 years of follow-up and 350 controls. Cases were matched to two controls on year of birth, date of blood draw, and fasting status. C-peptide levels were weakly associated with risk of colon cancer [top quartile (Q4) versus bottom quartile (Q1): multivariable relative risk (MVRR), 1.76; 95% confidence interval (95% CI), 0.85-3.63]. Fasting IGFBP-1 was inversely associated with risk of colon cancer (MVRR, 0.28; 95% CI, 0.11-0.75). We observed no clear association between glycosylated hemoglobin and risk for colorectal cancer. The IGF-I to IGFBP-3 molar ratio was associated with colon cancer risk (MVRR, 2.82; 95% CI, 1.35-5.88), and women with low levels of both IGF-I/IGFBP-3 and C-peptide (or high IGFBP-1) were at low risk, and elevation of either was sufficient to increase risk. Although altering IGF-I levels may not be practical, the growing burden of obesity and consequently hyperinsulinemia, which seems increasingly important for colon cancer, may be a target for effective prevention. PMID- 15824156 TI - Detection of alkaline sphingomyelinase activity in human stool: proposed role as a new diagnostic and prognostic marker of colorectal cancer. AB - OBJECTIVES: Intestinal alkaline sphingomyelinase, by exerting a major role in dietary sphingomyelin digestion, is responsible for the generation of messengers able to trigger the rapid turnover and apoptosis in intestinal epithelial cells. Markedly reduced mucosal alkaline sphingomyelinase activity has been associated with human colorectal neoplasms. The aim of this study was to analyze the alkaline sphingomyelinase activity in feces from healthy subjects and colorectal adenocarcinoma patients and to correlate it with the enzyme activity in intestinal tissues. MATERIALS AND METHODS: The enzyme activity was measured both in the intestinal samples from 12 healthy controls and 51 patients with colorectal adenocarcinoma (tumoral and paratumoral tissue) and in the fecal samples of 34 healthy subjects and 29 patients with adenocarcinoma. The relation between sphingomyelinase activity and Dukes' stage, cell differentiation degree, age, and gender was also analyzed. RESULTS: Alkaline sphingomyelinase was significantly decreased (P < 0.001; mean reduction >90%) in tumoral intestinal mucosa of patients compared with controls independently of Dukes' stage and tumor differentiation grade. Interestingly, the enzyme activity in histologically normal paratumoral tissues was statistically lower than control samples (P < 0.001). As occurs in neoplastic tissues, a relevant mean reduction (P < 0.0001; almost 90%) of alkaline sphingomyelinase was revealed in stool samples from tumor patients when compared with controls. CONCLUSION: These findings may have implications for cancer biology and perhaps also for the design of clinical test, thus suggesting that the fecal sphingomyelinase activity could really reflect the human intestinal mucosa enzyme level and could represent a new marker for human colorectal adenocarcinoma, mainly taking into account its early appearance in intestinal neoplasms. PMID- 15824157 TI - APC Asp1822Val and Gly2502Ser polymorphisms and risk of colorectal cancer and adenoma. AB - Mutation of the adenomatous polyposis coli (APC) tumor suppressor gene is an important initiating factor in the early stages of the adenoma-carcinoma sequence. The aim of this study was to investigate the two most common APC variants (Asp1822Val and Gly2502Ser) and their association with colorectal cancer and adenoma and whether these relationships are influenced by dietary and lifestyle factors. We analyzed 556 adenoma cases and 557 matched controls and 197 cancer cases and 490 matched controls nested within the Nurses' Health Study cohort, 274 cancer cases and 456 matched controls nested within the Physicians' Health Study cohort, and 375 adenoma cases and 724 matched controls nested within the Health Professionals Follow-up Study cohort. APC Asp1822Val and Gly2502Ser polymorphisms were not associated with risk of colorectal cancer or adenoma. For colorectal cancer, a significant interaction was found between Asp1822Val genotype and postmenopausal hormone (PMH) use among postmenopausal women (P(interaction) = 0.03). Current PMH use was associated with reduced risk overall and a statistically significant lower risk of colorectal cancer among carriers of one or two copies of the APC 1822Val allele (relative risk, 0.46; 95% confidence interval, 0.24-0.88) relative to wild-type never or past PMH users. Our results suggest that cigarette smoking, alcohol intake, and family history of colorectal cancer were positively associated and regular aspirin intake was inversely associated with colorectal adenoma in men and women. No gene-environment interactions were observed with these risk factors or with other dietary risk factors previously hypothesized to interact with the APC Asp1822Val polymorphism. PMID- 15824158 TI - The T393C polymorphism of the G alpha s gene (GNAS1) is a novel prognostic marker in bladder cancer. AB - The G protein G(alpha)s pathway is linked to proapoptotic signaling in cancer cell lines. To assess the role of the GNAS1 locus encoding G(alpha)s as a genetic factor for disease progression of transitional cell carcinoma (TCC) of the bladder, we genotyped the synonymous T393C polymorphism in 254 patients with TCC (minor allele frequency: 0.43) to examine a potential association between genotypes and disease progression. Using Kaplan-Meier estimates to calculate 5 year probabilities of follow-up, we could show that progression-free survival, metastasis-free survival, and cancer-specific survival was significantly increased in TT genotypes (56%, 84%, 82%) compared with CC genotypes (35%, 53%, 58%). In multivariate Cox proportional hazard analysis, the T393C polymorphism was an independent prognostic factor for clinical outcome. Homozygous CC patients were at highest risk for progression [odds ratio (OR), 1.94; P = 0.020], metastasis (OR, 3.49; P = 0.005), and tumor-related death (OR, 2.49; P = 0.031) compared with TT genotypes. Heterozygous patients had an intermediate risk compatible with a gene-dose effect. Real-time PCR analysis of urothelial tumor tissue as well as adipose and heart tissue revealed that G(alpha)s mRNA expression was highest in TT genotypes, indicating a proapoptotic effect in these genotypes. In conclusion, the GNAS1 T393C status associated with differential G(alpha)s mRNA expression is a novel independent prognostic marker for clinical outcome supporting a functional role of G(alpha)s in bladder cancer progression. PMID- 15824160 TI - Tobacco-specific nitrosamines and their pyridine-N-glucuronides in the urine of smokers and smokeless tobacco users. AB - Tobacco-specific nitrosamines are believed to play a significant role as causes of cancer in people who use tobacco products. Whereas the uptake of one tobacco specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, has been shown by analysis of its metabolites in urine, there are no published studies on urinary levels of N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT), and N' nitrosoanabasine (NAB) or their metabolites in human urine. We developed a method for quantitation of NNN, NAT, NAB, and their pyridine-N-glucuronides NNN-N-Gluc, NAT-N-Gluc, and NAB-N-Gluc in human urine. Total NNN (NNN plus NNN-N-Gluc) was assayed using 5-methyl-N'-nitrosonornicotine as internal standard. Urine was treated with beta-glucuronidase. Following solvent partitioning and solid-phase extraction, total NNN was determined using gas chromatography with nitrosamine selective detection. Total NAT and total NAB were quantified in the same samples. Separate quantitation of NNN and NNN-N-Gluc was accomplished by extraction of the urine with ethyl acetate before beta-glucuronidase hydrolysis; NNN was analyzed in the ethyl acetate extract, and after enzyme treatment, NNN released from NNN-N Gluc was quantified in the extracted urine. Separate analyses of NAT, NAT-N-Gluc, NAB, and NAB-N-Gluc proceeded similarly. Analyte identities were confirmed by gas chromatography-tandem mass spectrometry. Mean levels of total NNN, NAT, and NAB in the urine of 14 smokers were (pmol/mg creatinine) 0.18 +/- 0.22, 0.19 +/- 0.20, and 0.040 +/- 0.039, respectively, whereas the corresponding amounts in the urine of 11 smokeless tobacco users were 0.64 +/- 0.44, 1.43 +/- 1.10, and 0.23 +/- 0.19, respectively. Pyridine-N-glucuronides accounted for 59% to 90% of total NNN, NAT, and NAB. The results of this study show the presence of NNN, NAT, NAB, and their pyridine-N-glucuronides in human urine and provide a quantitative method for application in mechanistic and epidemiologic studies of the role of tobacco-specific nitrosamines in human cancer. PMID- 15824159 TI - Polymorphisms in XPD exons 10 and 23 and bladder cancer risk. AB - INTRODUCTION: The nucleotide excision repair gene, xeroderma pigmentosum complementation group D (XPD), has been hypothesized to have a role in cancer risk, but results from prior molecular epidemiologic studies and genotype phenotype analyses are conflicting. MATERIALS AND METHODS: We examined the frequency of the XPD Asp312Asn polymorphism in exon 10 and the XPD Lys751Gln polymorphism in exon 23 in 505 incident bladder cancer cases and 486 healthy controls. RESULTS: Overall, the XPD exon 10 and 23 genotypes were not associated with bladder cancer risk, after adjusting for age, sex, ethnicity, and smoking status. A gender-specific role was evident that showed an increased risk for women, but not for men, associated with the variant genotypes for both exons. For example, when the exon 23 variant allele genotypes were combined (Lys/Gln + Gln/Gln), there was an increased bladder cancer risk in women [odds ratio (OR), 1.69; 95% confidence interval (95% CI), 1.12-2.58] but not in men (OR, 0.99; 95% CI, 0.79-1.24; P(interaction) = 0.041; OR, 1.62; 95% CI, 1.02-2.58). There was also a gene-smoking interaction that showed the variant alleles for either exon or the combination of both increase the risk of bladder cancer for light and heavy smokers. For exon 23 (P(interaction) = 0.057; OR, 1.21; 95% CI, 0.99-1.47), heavy smokers (> or = 20 pack-years) who carried the exon 23 variant allele genotypes had an OR of 4.13 (95% CI, 2.53-6.73), whereas heavy smokers with the wild-type genotypes were at lower risk (OR, 3.55; 95% CI, 2.19-5.75). Moderate smokers (1-19 pack-years) with the variant allele genotypes had an OR of 1.54 (95% CI, 0.94-2.53), whereas moderate smokers with the wild-type genotypes had an OR of 1.12 (95% CI, 0.63-1.98). CONCLUSIONS: Although we did not observe main effects associated with the XPD genotypes, these results do suggest the variant allele genotypes were associated with increased bladder cancer risk in women and smokers with statistically significant interactions in the exon 23 polymorphism. Although there is biological plausibility, these novel findings for gender and smoking should be interpreted with caution upon confirmation in larger studies. PMID- 15824161 TI - Safety and efficacy of weekly oral oltipraz in chronic smokers. AB - Cigarette smoking is thought to contribute to carcinogenesis by formation of DNA adducts of tobacco smoke constituents leading to genotoxic damage. The dithiolethione, oltipraz, is a putative cancer chemopreventive agent that induces phase II detoxifying enzymes in preclinical models and reduces aflatoxin adducts in humans living in areas with high dietary levels. To determine if oltipraz could reduce adduct levels of tobacco smoke constituents in the lungs and other target organs, chronic smokers were enrolled to one of three arms: 400 or 200 mg/wk oral oltipraz or placebo. Endobronchial tissue and bronchoalveolar lavage were done before and after 12 weeks of drug treatment; peripheral blood, urine, and oral saline rinse were also collected. Toxicity was assessed every 4 weeks. Fifty-nine of the 77 enrolled subjects completed the study. Of those receiving oltipraz, 15% experienced grade 2/3 toxicity, which was predominantly gastrointestinal. All subject withdrawals occurred in the oltipraz groups. There was no significant difference between pre- and post-polycyclic aromatic hydrocarbon-DNA adduct levels in lung epithelial cells measured by immunoperoxidase staining between treatment and placebo groups. Likewise, no significant differences were found in polycyclic aromatic hydrocarbon or benzo(a)pyrene-7,8-diol-9,10-epoxide adducts measured in blood, oral lining cells, or bladder lining cells. There was also no increase in mRNA or enzymatic activity of phase II enzymes and no change in glutathione levels. Thus, despite moderate drug-related toxicity, there was no significant effect on pharmacodynamic or surrogate risk biomarkers. Other agents with lower toxicity and greater activity to induce phase II enzymes are needed to definitively test the detoxification-induction paradigm in smokers. PMID- 15824162 TI - Fatty acid composition of red blood cell membranes and risk of squamous cell carcinoma of the skin. AB - Differential effects of fatty acids on carcinogenesis suggest that fatty acid composition is important in tumor development. Arachidonic acid and its metabolites elicit inflammation and promote tumor formation in mouse skin. Inhibitors of the arachidonic cascade inhibit tumor incidence. A population-based case control study in Southeastern Arizona tested the hypothesis that lower levels of arachidonic acid in RBC membranes were associated with decreased risk of skin squamous cell carcinoma (SCC; n = 335 SCC cases and 321 controls). Extracted and esterified RBC fatty acids were analyzed using capillary gas chromatography. Individual peaks for 14 fatty acids were measured as a percentage of total fatty acids. Logistic regression was used to estimate odds ratios (OR), adjusting for SCC risk factors (age, gender, actinic keratosis history, freckling, and tanning ability). Increased levels of arachidonic acid in RBC membranes were associated with increased risk of SCC [odds ratio (OR), 1.08 per mg/100 mL change; 95% confidence interval (95% CI), 1.02-1.15] and this association remained when controls with actinic keratosis precursor lesions were excluded. SCC risk was highest among the upper quartile of arachidonic acid (OR, 2.38; 95% CI, 1.37-4.12). In contrast, increasing proportions of palmitic acid (OR, 0.94; 95% CI, 0.89-1.00) and palmitoleicacid (OR, 0.49; 95% CI, 0.30-0.81) were associated with reduced SCC risk. More studies are needed to elucidate the function of RBC fatty acids so that recommendations can be made to alter the human diet for cancer prevention. PMID- 15824163 TI - No Evidence for BRAF as a melanoma/nevus susceptibility gene. AB - Somatic mutations of BRAF have been identified in both melanoma tumors and benign nevi. Germ line mutations in BRAF have not been identified as causal in families predisposed to melanoma. However, a recent study suggested that a BRAF haplotype was associated with risk of sporadic melanoma in men. Polymorphisms or other variants in the BRAF gene may therefore act as candidate low-penetrance genes for nevus/melanoma susceptibility. We hypothesized that promoter variants would be the most likely candidates for determinants of risk. Using denaturing high pressure liquid chromatography and sequencing, we screened peripheral blood DNA from 184 familial melanoma cases for BRAF promoter variants. We identified a promoter insertion/deletion in linkage disequilibrium with the previously described BRAF polymorphism in intron 11 (rs1639679) reported to be associated with melanoma susceptibility in males. We therefore investigated the contribution of this BRAF polymorphism to melanoma susceptibility in 581 consecutively recruited incident cases, 258 incident cases in a study of late relapse, 673 female general practitioner controls, and the 184 familial cases. We found no statistically significant difference in either genotype or allele frequencies between cases and controls overall or between male and female cases for the BRAF polymorphism in the two incident case series. Our results therefore suggest that the BRAF polymorphism is not significantly associated with melanoma and the promoter insertion/deletion linked with the polymorphism is not a causal variant. In addition, we found that there was no association between the BRAF genotype and mean total number of banal or atypical nevi in either the cases or controls. PMID- 15824164 TI - Intraindividual variability in arsenic methylation in a U.S. population. AB - Several recent investigations have reported associations between a reduced capacity to fully methylate inorganic arsenic and increased susceptibility to arsenic-caused cancer. In these studies, methylation patterns were based on a single assessment of urinary arsenic metabolites collected at the time of cancer diagnosis. However, the latency of arsenic-caused cancer may be several decades, and the extent to which a recent measurement can be used to estimate a person's past methylation pattern is unknown. In this investigation, the distribution of urinary inorganic arsenic, monomethylarsonate, and dimethylarsinate was used to assess intraindividual variation in methylation capacity in 81 subjects with low to moderate arsenic exposures. Multiple urine samples were collected from each subject over a 1-year period. Duplicate analyses done on 27 samples were used to assess laboratory measurement imprecision. The intraclass correlation coefficients (ICC) for the proportion of urinary arsenic as inorganic arsenic, monomethylarsonate, and dimethylarsinate in samples taken an average of 258 days apart, were 0.45 [95% confidence interval (95% CI), 0.23-0.63] 0.46 (95% CI, 0.24 0.64), and 0.49 (95% CI, 0.28-0.66). In analyses of duplicate samples, ICCs for the concentration of arsenic species ranged from 0.87 to 0.93, whereas ICCs for species proportions ranged from 0.63 to 0.76. These data suggest that individual methylation patterns remain fairly stable over time, although variability due to measurement imprecision or intraindividual changes over time does occur. This variability could lead to misclassification of methylation patterns and could bias relative risk estimates in studies of methylation and cancer towards the null. PMID- 15824165 TI - Cigarette smoking and risk of non-Hodgkin lymphoma: a pooled analysis from the International Lymphoma Epidemiology Consortium (interlymph). AB - BACKGROUND: The International Lymphoma Epidemiology Consortium (InterLymph) provides an opportunity to analyze the relationship between cigarette smoking and non-Hodgkin lymphoma with sufficient statistical power to consider non-Hodgkin lymphoma subtype. The results from previous studies of this relationship have been inconsistent, likely due to the small sample sizes that arose from stratification by disease subtype. To clarify the role of cigarette smoking in the etiology of non-Hodgkin lymphoma, we conducted a pooled analysis of original patient data from nine case-control studies of non-Hodgkin lymphoma conducted in the United States, Europe, and Australia. METHODS: Original data were obtained from each study and uniformly coded. Risk estimates from fixed-effects and two stage random-effects models were compared to determine the impact of interstudy heterogeneity. Odds ratios (OR) and 95% confidence intervals (95% CI) were derived from unconditional logistic regression models, controlling for study center, age, sex, and race. RESULTS: In our pooled study population of 6,594 cases and 8,892 controls, smoking was associated with slightly increased risk estimates (OR, 1.07; 95% CI, 1.00-1.15). Stratification by non-Hodgkin lymphoma subtype revealed that the most consistent association between cigarette smoking and non-Hodgkin lymphoma was observed among follicular lymphomas (n = 1452). Compared with nonsmokers, current smokers had a higher OR for follicular lymphoma (1.31; 95% CI, 1.12-1.52) than former smokers (1.06; 95% CI, 0.93-1.22). Current heavy smoking (> or = 36 pack-years) was associated with a 45% increased OR for follicular lymphoma (1.45; 95% CI, 1.15-1.82) compared with nonsmokers. CONCLUSIONS: Cigarette smoking may increase the risk of developing follicular lymphoma but does not seem to affect risk of the other non-Hodgkin lymphoma subtypes we examined. Future research is needed to determine the biological mechanism responsible for our subtype-specific results. PMID- 15824166 TI - Residential herbicide use and risk of non-Hodgkin lymphoma. AB - CONTEXT: Environmental exposure to herbicides has been hypothesized to contribute to the long-term increase in non-Hodgkin lymphoma (NHL). OBJECTIVE: To estimate the effects of residential herbicide exposure on NHL risk. DESIGN: Population based case-control study. SETTING: Iowa and metropolitan Detroit, Los Angeles, and Seattle, 1998 to 2000. PARTICIPANTS: NHL patients ages 20 to 74 years and unaffected residents identified by random digit dialing and Medicare eligibility files. MAIN OUTCOME MEASURES: Computer-assisted personal interviews (1,321 cases, 1,057 controls) elicited data on herbicide use at each home occupied since 1970. Levels of 2,4-dichlorophenoxy-acetic acid and dicamba were measured in dust taken from used vacuum cleaner bags in the current home (679 cases, 510 controls who had owned at least half of their carpets for > or = 5 years). RESULTS: Herbicide use on the lawn or garden was similar among cases and controls (adjusted relative risk, 1.02; 95% confidence interval, 0.84-1.23). Estimated risk did not increase with greater duration, frequency, or total number of applications of herbicides to the lawn, the garden, or to both combined. Risk was not elevated for respondents who applied the herbicides themselves and not for those exposed during the 1970s, 1980s, or 1990s. We detected 2,4-dichlorophenoxy-acetic acid equally often in homes of cases and controls (78%). We found dicamba in homes of 15% of cases and 20% of controls. We also found no elevation in risk among the respondents who had the highest dust levels and highest self-reported exposures. We found no consistent patterns for specific histologies. CONCLUSIONS: We found no detectable excess associated with residential exposures. Residential herbicide exposures are unlikely to explain the long-term increase in NHL. PMID- 15824167 TI - The MTHFR 1298A>C polymorphism and genomic DNA methylation in human lymphocytes. AB - Methylenetetrahydrofolate reductase (MTHFR) balances the pool of folate coenzymes in one-carbon metabolism for DNA synthesis and methylation, both implicated in carcinogenesis. Epidemiologic studies have shown that two functional polymorphisms in MTHFR gene, 677C>T and 1298A>C, are related to increased cancer risk. We aimed to analyze lymphocyte DNA from 198 subjects to evaluate the MTHFR 1298A>C polymorphism and folate status affecting genomic DNA methylation as a possible mechanism underlying the relationship between MTHFR polymorphisms and cancer susceptibility. Carriers of the 1298AA wild-type genotype showed lower genomic DNA methylation compared with 1298AC or 1298CC genotypes [3.72 versus 8.59 or 6.79 ng 5-methyl-2'-deoxycytidine (5-mCyt)/microg DNA, P < 0.0001 and P = 0.007, respectively]. When DNA methylation was evaluated according to plasma folate status, only 1298AA with low folate levels revealed diminished DNA methylation (P < 0.0001). Moreover, when the two MTHFR polymorphisms were concomitantly evaluated at the low folate status, DNA methylation was reduced only in 1298AA/677TT compared with 1298AA/677CC (3.11 versus 7.29 ng 5 mCyt/microg DNA, P = 0.001) and 1298CC/677CC genotypes (3.11 versus 7.14 ng 5 mCyt/microg DNA, P = 0.004). However, the high prevalence of 677TT mutants within the 1298AA group (79%) and the similar biochemical features of 1298AA/677CC and 1298CC/677CC combined genotypes suggest that the gene-nutrient interaction affecting DNA methylation in 1298AA is mainly due to the coexistence of the 677TT genotype and that the 1298A>C polymorphism may convey its protective effect not through this interaction but through another pathway in one-carbon metabolism. Further mechanistic studies are warranted to investigate how single polymorphisms as well as MTHFR combined genotypes exert their effect on cancer susceptibility. PMID- 15824168 TI - Dietary folate and risk of prostate cancer in Italy. AB - Folate status may affect cancer risk through its role in both methylation and nucleotide synthesis of DNA. A low dietary intake of folate has been linked to risk of several cancers, but epidemiologic studies with reference to prostate cancer are scanty. We therefore analyzed data from a case-control study of prostate cancer conducted between 1991 and 2002 in various areas of Italy. Cases were 1,294 patients with incident, histologically confirmed prostate cancer and controls were 1,451 patients admitted to the same network of hospitals of cases for acute, nonneoplastic conditions. All subjects were < 75 years old. Intake of folate and other nutrients was computed from a validated food frequency questionnaire. We adjusted for energy intake using the residual method, and calculated multivariate odds ratios (OR) and 95% confidence intervals (CI) using unconditional logistic regression. The OR of prostate cancer was 0.66 (95% CI, 0.51-0.85) for the highest versus the lowest quintile of folate intake. The relation between dietary folate and prostate cancer was consistent across strata of age, methionine, vitamin B6, and alcohol intake, and did not vary substantially according to Gleason score of prostate cancer. The combined OR for high-folate and low-alcohol intake versus low-folate and high-alcohol intake was 0.46 (95% CI, 0.29-0.75). Therefore, this study supports a favorable role of dietary folate on prostate cancer risk. PMID- 15824169 TI - Association of susceptibility alleles in ELAC2/HPC2, RNASEL/HPC1, and MSR1 with prostate cancer severity in European American and African American men. AB - Reported associations of ELAC2/HPC2, RNASEL/HPC1, and MSR1 with prostate cancer have been inconsistent and understudied in African Americans. We evaluated the role of 16 sequence variants in these genes with prostate cancer using 888 European American and 131 African American cases, and 473 European American and 163 African American, controls. We observed significant differences in ELAC2, RNASEL, and MSR1 allele frequencies by race. However, we did not observe significant associations between prostate cancer and any variants examined for both races combined. Associations were observed when stratified by race, family history, or disease severity. European American men homozygous for MSR1 IVS7delTTA had an elevated risk for localized stage [odds ratio, (OR), 3.5; 95% confidence interval (95% CI), 1.4-6.9], low-grade (OR, 3.2; 95% CI, 1.4-7.3) disease overall, and with low-grade (OR, 2.9; 95% CI, 1.2-7.2) or late-stage disease (OR, 5.2; 95% CI, 1.1-25.7) in family history-negative African Americans. MSR1 Arg293X was associated with family history-negative high-grade disease (OR, 4.0; 95% CI, 1.1-14.1) in European Americans. RNASEL Arg462Gln was associated with low-grade (OR, 1.5; 95% CI, 1.04-2.2) and early-stage (OR, 1.5; 95% CI, 1.02 2.1) disease in family history-negative European Americans. In family history positive individuals, Arg462Gln was inversely associated with low-grade (OR, 0.43; 95% CI, 0.21-0.88) and low-stage (OR, 0.46; 95% CI, 0.22-0.95) disease. In African Americans, Arg462Gln was associated with positive family history high stage disease (OR, 14.8; 95% CI, 1.6-135.7). Meta-analyses revealed significant associations of prostate cancer with MSR1 IVS7delTTA, -14,742 A>G, and Arg293X in European Americans; Asp174Tyr in African Americans; RNASEL Arg462Gln in European American's overall and in family history-negative disease; and Glu265X in family history-positive European Americans. Therefore, MSR1 and RNASEL may play a role in prostate cancer progression and severity. PMID- 15824170 TI - A DNA pooling strategy for family-based association studies. AB - Genome-wide association scans for disease susceptibility genes of complex diseases require genotyping on a massive scale. A DNA pooling strategy for family based association studies is described, which is robust to population stratification biases and to errors in pooling. It can achieve a statistical efficiency of 0.95 with approximately 1 of 8 or fewer genotyping efforts, and an efficiency of 0.90 with approximately 1 of 16 or fewer efforts compared with individual genotyping. The pooling method described in this article provides a tradeoff between genotyping efforts and subject recruitment efforts. PMID- 15824171 TI - Lifestyle determinants and mortality in German vegetarians and health-conscious persons: results of a 21-year follow-up. AB - BACKGROUND: The long-term observation of vegetarians in affluent countries can provide insight into the relative effects of a vegetarian diet and lifestyle factors on mortality. METHODS: A cohort study of vegetarians and health-conscious persons in Germany was followed-up prospectively for 21 years, including 1,225 vegetarians and 679 health-conscious nonvegetarians. Standardized mortality ratios compared with the German general population were calculated for all causes and specific causes. Within the cohort, Poisson regression modeling was used to investigate the joint effects of several risk factors on overall and cause specific mortality. RESULTS: Standardized mortality ratios for all-cause mortality was significantly below 100: 59 [95% confidence interval (95% CI), 54 64], predominantly due to a deficit of deaths from circulatory diseases. Within the cohort, vegetarian compared with nonvegetarian diet had no effect on overall mortality [rate ratio (RR), 1.10; 95% CI, 0.89-1.36], whereas moderate and high physical activity significantly reduced risk of death (RR, 0.62, 0.64), adjusted for age, sex, smoking, alcohol intake, body mass index, and educational level. Vegetarian diet was however associated with a reduced RR of 0.70 (95% CI, 0.41 1.18) for ischemic heart disease, which could partly be related to avoidance of meat. CONCLUSIONS: Both vegetarians and nonvegetarian health-conscious persons in this study have reduced mortality compared with the general population. Within the study, low prevalence of smoking and moderate or high level of physical activity but not strictly vegetarian diet was associated with reduced overall mortality. The nonsignificant reduction in mortality from ischemic heart diseases in vegetarians compared with health-conscious persons could be explained in part by avoidance of meat intake. PMID- 15824172 TI - Genotyping of patients with sporadic and radiation-associated meningiomas. AB - Ionizing radiation is the most established risk factor for meningioma formation. Our aim was to evaluate the main effect of selected candidate genes on the development of meningioma and their possible interaction with ionizing radiation in the causation of this tumor. The total study population included 440 cases and controls: 150 meningioma patients who were irradiated for tinea capitis in childhood, 129 individuals who were similarly irradiated but did not develop meningioma, 69 meningioma patients with no previous history of irradiation, and 92 asymptomatic population controls. DNA from peripheral blood samples was genotyped for single nucleotide polymorphisms (SNP) in 12 genes: NF2, XRCC1, XRCC3, XRCC5, ERCC2, Ki-ras, p16, cyclin D1, PTEN, E-cadherin, TGFB1, and TGFBR2. SNP analysis was done using the MassArray system (Sequenom, San Diego, CA) and computerized analysis by SpectroTYPER. Logistic regressions were applied to evaluate main effect of each gene on meningioma formation and interaction between gene and radiation. Intragenic SNPs in the Ki-ras and ERCC2 genes were associated with meningioma risk (odds ratio, 1.76; 95% confidence interval, 1.07-2.92 and odds ratio, 1.68; 95% confidence interval, 1.00-2.84, respectively). A significant interaction was found between radiation and cyclin D1 and p16 SNPs (P for interaction = 0.005 and 0.057, respectively). Our findings suggest that Ki ras and ERCC2 SNPs are possible markers for meningioma formation, whereas cyclin D1 and p16 SNPs may be markers of genes that have an inverse effect on the risk to develop meningioma in irradiated and nonirradiated populations. PMID- 15824173 TI - The effect of a school-based sun protection intervention on the development of melanocytic nevi in children: 6-year follow-up. AB - Because nevi share risk factors with melanoma and are strong risk factors for melanoma, they are suitable biomarkers for evaluating sun protection programs. Kidskin was a trial of a school-based sun protection program in Western Australia that included high and moderate intervention groups and a control group. Schools were assigned nonrandomly to groups. The primary outcome was number of nevi on the back. Nevi were counted at baseline, after 4 years intervention and again 2 years later. Linear growth models, allowing for correlated data within schools and children were fitted to the data. The primary analysis included 639 control children, 414 in the moderate and 355 in the high intervention group. Compared with the control group, the relative increase in number of nevi on the back was 0.89 (95% confidence interval, 0.81-0.99) for the high intervention group and 0.94 (95% confidence interval, 0.86-1.04) for the moderate group (P = 0.09). In subgroup analyses of nevi on the back, the association was stronger in boys (P < 0.001) than in girls (P = 0.7), although the test for interaction was not significant (P = 0.11). For the chest, examined in boys, the associations were similar to that for nevi on the back in boys. Associations were weak for nevi on the face and arms (P = 0.2); for this site, there was weak evidence of heterogeneity by sex. Overall, there was weak evidence that the Kidskin intervention reduced the number of new nevi over a 6-year period, but there was stronger evidence of an effect on the trunk in boys. PMID- 15824174 TI - Multiplexed immunobead-based cytokine profiling for early detection of ovarian cancer. AB - Early detection of ovarian cancer might improve clinical outcome. Some studies have shown the role of cytokines as a new group of tumor markers for ovarian cancer. We hypothesized that a panel comprised of multiple cytokines, which individually may not show strong correlation with the disease, might provide higher diagnostic power. To evaluate the diagnostic utility of cytokine panel, we used a novel multianalyte LabMAP profiling technology that allows simultaneous measurement of multiple markers. Concentrations of 24 cytokines (cytokines/chemokines, growth, and angiogenic factors) in combination with cancer antigen-125 (CA-125), were measured in sera of 44 patients with early-stage ovarian cancer, 45 healthy women, and 37 patients with benign pelvic tumors. Six markers, i.e., interleukin (IL)-6, IL-8, epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), and CA-125, showed significant differences in serum concentrations between ovarian cancer and control groups. Out of this group, IL-6, IL-8, VEGF, EGF, and CA-125, were used in a classification tree analysis that resulted in 84% sensitivity at 95% specificity. The receiver operator characteristic curve created using the combination of markers produced sensitivities between 90% and 100% in the area of 80% to 90% specificity, whereas the receiver operator characteristic curve for CA 125 alone resulted in sensitivities of 70% to 80%. The classification tree analysis for discrimination of benign condition from ovarian cancer used CA-125, granulocyte colony-stimulating factor (G-CSF), IL-6, EGF, and VEGF resulting in 86.5% sensitivity and 93.0% specificity. The presented data show that simultaneous testing of a panel of serum cytokines and CA-125 using LabMAP technology may present a promising approach for ovarian cancer detection. PMID- 15824175 TI - Serum trans-fatty acids are associated with risk of prostate cancer in beta Carotene and Retinol Efficacy Trial. AB - Biomarkers of trans-fatty acid consumption have been associated with increased risks of breast and colon cancer, although no studies have examined their associations with prostate cancer risk. Using data from the beta-Carotene and Retinol Efficacy Trial, this nested case-control study examined the relationships between serum phospholipid trans-fatty acids and prostate cancer incidence in 272 case and 426 control men. Trans-fatty acids were measured using organic extraction followed by separations with TLC and gas chromatography. Adjusted odds ratios for risk of prostate cancer with increasing levels of trans-fatty acids were calculated using logistic regression. There were consistent trends for increasing prostate cancer risk with higher levels of C18 but not C16 trans-fatty acids, although only trends for Delta11t 18:1 trans-vaccenic and Delta9c,12t 18:2 fatty acids reached statistical significance. Odds ratios (95% confidence interval) contrasting low versus high quartiles for these fatty acids were 1.69 (1.03-2.77) and 1.79 (1.02-3.15), respectively. There were no consistent differences in associations between low-grade and high-grade cancer among the subset of 209 cases with information on tumor grade. Additional studies are needed to confirm these findings and better control for factors, such as use of prostate-specific antigen screening, which may confound this association. PMID- 15824176 TI - The E211 G>A androgen receptor polymorphism is associated with a decreased risk of metastatic prostate cancer and androgenetic alopecia. AB - The androgen receptor (AR) gene encodes a transcription factor, which mediates androgen action in target tissues, including the prostate. Prostate cancer is androgen dependent, implicating AR in susceptibility to this male condition. Male pattern balding, androgenetic alopecia, has recently been associated with prostate cancer, suggesting shared androgen pathways. The CAG and GGC repeats in the AR have been studied extensively as markers of prostate cancer susceptibility, with inconclusive findings, whereas the AR-E211 G>A polymorphism has been associated with androgenetic alopecia. We assessed the repeat linked single nucleotide polymorphism as a marker of risk association in prostate cancer, including androgenetic alopecia, in an Australian population-based case control study. In 815 prostate cancer cases and 719 controls, the proportion of A allele carriers was the same in each group. Overall, there was no evidence for an association between the A allele and risk of prostate cancer, however, the proportion of A-allele carriers in metastatic prostate cancer (5%) was lower than in less advanced disease (16%, P = 0.03). The proportion of A-allele carriers was 24% in nonbald men but it was lower in men with vertex alopecia alone (13%, P = 0.001) or in combination with frontal alopecia (7%, P < 0.0001). This inverse association between the A allele and baldness was independent of prostate cancer status (P for interaction = 0.2). These results suggest that the AR-E211 A allele, in linkage with the functional repeat sequences, is associated with a lower risk of metastatic prostate cancer and a lower risk of alopecia. PMID- 15824177 TI - Genetic variants in the vitamin D receptor gene and prostate cancer risk. AB - Vitamin D receptor (VDR), a member of the steroid/thyroid hormone nuclear receptor family, is bound by the steroid hormone 1,25-dihydroxyvitamin D3, which is thought to play a role in the etiology and progression of prostate cancer. Polymorphisms in the VDR gene have been associated with prostate cancer risk, although findings are inconclusive. The purpose of this study was to determine if VDR polymorphisms were associated with prostate cancer risk using a large, Australian population-based study of 812 cases and 713 controls frequency-matched by age. As the 3' region polymorphisms are in strong linkage disequilibrium, for joint effects, we only evaluated the common g.60890G > A polymorphism with the unlinked g.27823C > T (5' region) polymorphism. Allele frequencies were similar in cases and controls (g.27823C > T, 36% versus 36%; g.60890 G>A, 41% versus 43%). No genotypes were individually associated with prostate cancer risk (all P > 0.3). All nine possible genotype combinations were evident, and although the g.27823CT/g.60890GA combination was nominally more prevalent in controls (24%) than in cases (19%, P = 0.03), there was no difference in the combined genotype distribution between cases and controls (P = 0.2). The associations of risk with genotype were between 0.91 and 1.03, all with 95% confidence intervals within 0.81 to 1.15. In conclusion, VDR polymorphisms either alone or in combination do not seem to contribute appreciably to prostate cancer risk. PMID- 15824178 TI - Levels of physical activity for colon cancer prevention compared with generic public health recommendations: population prevalence and sociodemographic correlates. AB - BACKGROUND: The proportion of Australian adults achieving physical activity levels believed to be sufficient for colon cancer prevention was estimated, and sociodemographic correlates (age, gender, educational attainment, occupation, marital status, and children in household) of meeting these levels of activity were analyzed. METHODS: Data from the 2000 National Physical Activity Survey were used to estimate the prevalence of participation in physical activity in relation to three criteria: generic public health recommendations, weekly amount of at least moderate-intensity physical activity currently believed to reduce risk of colon cancer, and weekly amount of vigorous-intensity physical activity believed to reduce risk of colon cancer. RESULTS: Overall, 46% of adults met the generic public health criterion, 26% met the colon cancer criterion based on participation in at least moderate-intensity physical activity, and 10% met the colon cancer criterion based on vigorous-intensity physical activity. Women were less likely than men to meet the colon cancer criteria. Younger and more educated persons were more likely to meet all three criteria. The most pronounced differences between gender, age, and educational attainment groups were found for meeting the amount of vigorous-intensity physical activity believed to reduce risk of colon cancer. CONCLUSIONS: The population prevalence for meeting proposed physical activity criteria for colon cancer prevention is low and much lower than that related to the more generic public health recommendations. If further epidemiologic studies confirm that high volumes and intensities of activity are required, the public health challenges for colon cancer will be significant. PMID- 15824179 TI - Utilization of BRCA1/BRCA2 mutation testing in newly diagnosed breast cancer patients. AB - BACKGROUND: Among newly diagnosed breast cancer patients who are at risk for carrying a BRCA1 or BRCA2 mutation, knowledge of mutation status can influence local breast cancer treatment decisions. Thus, genetic testing at the time of diagnosis is increasingly considered an option for such patients. In this study, we evaluated factors associated with the decision to undergo BRCA1/BRCA2 gene testing at the time of initial breast cancer diagnosis. METHODS: Participants were newly diagnosed breast cancer patients who had not yet received definitive local breast cancer treatment and who had a family history consistent with hereditary breast cancer. Participants were offered genetic counseling and BRCA1/BRCA2 testing with results in 2 to 3 weeks. RESULTS: Of 231 patients who referred to the study, 20 (9%) declined the baseline interview, 34 (15%) completed a baseline interview but declined genetic testing, and 177 (76%) underwent BRCA1/BRCA2 testing. Physician recommendation for BRCA1/BRCA2 testing and indecision about definitive local treatment were both associated with undergoing testing. Among patients who were tested, 38 (21%) proceeded with definitive local treatment before receiving test results. Delay in the availability of test results and low levels of anxiety were associated with the decision to proceed with definitive local treatment before receiving test results. CONCLUSIONS: These results suggest that if rapid testing is available and genetic referrals are made for appropriate patients, a high proportion are likely to opt for such testing. In particular, patients who have not yet reached a decision about definitive local treatment may benefit from a genetic referral. PMID- 15824180 TI - Age-specific incidence rates of in situ breast carcinomas by histologic type, 1980 to 2001. AB - Incidence rates of ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS) have increased rapidly over the past several decades largely due to the increased use of mammography. However, recent data from 1987 to 1999 indicate that invasive ductal carcinoma incidence rates have remained essentially constant, whereas rates of invasive lobular carcinoma have increased 65%, with greater increases observed among postmenopausal women. Data on recent trends in DCIS and LCIS incidence rates, particularly age-specific trends, are lacking. We evaluated trends in the incidence rates of DCIS overall, noncomedo DCIS, comedo DCIS, and LCIS using data from nine Surveillance, Epidemiology, and End Results cancer registries. DCIS incidence rates increased 7.2-fold [95% confidence interval (95% CI), 6.8-7.7] from 1980 to 2001, 1.8-fold (95% CI, 1.7-1.9) over the past 10 years (1992-2001), and 1.1-fold (95% CI, 1.0-1.2) over the past 5 years (1997-2001). The magnitudes of these increases were highest among women ages > or = 50 years. Furthermore, over the past 10- and 5-year periods, rates of noncomedo DCIS have generally increased across all age groups, whereas rates of comedo DCIS held constant or decreased. LCIS incidence rates increased 2.6-fold (95% CI, 2.3-2.9) from 1980 to 2001, 1.3-fold (95% CI, 1.2-1.5) over the past 10 years, and 1.1-fold (95% CI, 1.0-1.3) over the past 5 years. Similar to invasive lobular carcinoma, but unlike invasive ductal carcinoma, incidence rates of both DCIS and LCIS continue to increase in the United States primarily among older women. These trends present important public health and clinical challenges. PMID- 15824181 TI - Cimetidine and other histamine2-receptor antagonist use in relation to risk of breast cancer. AB - OBJECTIVE: Cimetidine, a histamine2-receptor antagonist (H2 blocker) commonly used to treat symptoms of peptic and duodenal ulcer, influences both hormonal and immune pathways. We investigated the influence of cimetidine use on the risk of breast cancer in our hospital-based case control surveillance study. METHODS: Data on medication use and other factors were elicited from patients admitted to hospitals from 1977 to 2002. We compared 6,994 breast cancer cases with a control group comprising cancer (n = 2,478) and noncancer (n = 6,004) diagnoses. Conditional logistic regression models were used to estimate odds ratios for H2 blocker use that began at least 1 year prior to admission. Regular use was defined as use for at least 4 days per week for at least 3 continuous months. RESULTS: The odds ratio for breast cancer among regular users of cimetidine was 0.9 (95% confidence interval, 0.6-1.2) using a combined cancer and noncancer control group. For use of 4 or more years' duration, the odds ratio was < 1.0 but was not statistically significant. The odds ratio for the regular use of other H2 blockers was 0.9 (95% confidence interval, 0.6-1.3). CONCLUSIONS: Our data agree with data from three prior studies which indicate that cimetidine is not associated with the risk of breast cancer. Other H2 blockers were also unrelated to the risk of breast cancer. PMID- 15824182 TI - Effects of electron-beam irradiation on whole genome amplification. AB - Electron-beam (E-beam) irradiation, currently being used to sterilize mail addressed to selected ZIP codes in the United States, has significant negative effects on the genomic integrity of DNA extracted from buccal-cell washes. We investigated the yield, composition, and genotyping performance of whole genome amplified DNA (wgaDNA) derived from 24 matched samples of E-beam-irradiated and nonirradiated genomic DNA (gDNA) as a model for the effects of degraded gDNA on the performance of whole genome amplification. gDNA was amplified using the Multiple Displacement Amplification method. Three methods of DNA quantification analysis were used to estimate the yield and composition of wgaDNA, and 65 short tandem repeat and single nucleotide polymorphism genotyping assays were used to evaluate the genotyping performance of irradiated and nonirradiated gDNA and wgaDNA. Compared with wgaDNA derived from nonirradiated gDNA, wgaDNA derived from irradiated gDNA exhibited a significantly reduced yield of wgaDNA and significantly reduced short tandem repeat and single nucleotide polymorphism genotyping completion and concordance rates (P < 0.0001). Increasing the amount of irradiated gDNA input into whole genome amplification improved genotyping performance of wgaDNA but not to the level of wgaDNA derived from nonirradiated gDNA. Multiple Displacement Amplification wgaDNA derived from E-beam-irradiated gDNA is not suitable for genotyping analysis. PMID- 15824183 TI - No effect of exercise on insulin-like growth factor 1 and insulin-like growth factor binding protein 3 in postmenopausal women: a 12-month randomized clinical trial. PMID- 15824184 TI - NO excess risk of breast cancer among female users of systemic glucocorticoids. PMID- 15824185 TI - No association of meat, fish, and egg consumption with ovarian cancer risk. PMID- 15824186 TI - Reported use of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors was not associated with reduced recurrence of colorectal adenomas. AB - We did a secondary analysis of data from three large colorectal adenoma chemoprevention trials to assess the association between 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor use and reduced risk of recurrent colorectal adenomas. Reported use of HMG-CoA reductase inhibitors was not associated with a reduced recurrence of colorectal adenomas, multiple adenomas, or advanced adenomas. Lack of statistical power from limited exposure to HMG-CoA reductase inhibitors might be responsible for the lack of association. PMID- 15824187 TI - No association between cytochrome P450 and glutathione S-transferase gene polymorphisms and risk of colorectal adenoma: results from the UK Flexible Sigmoidoscopy Screening Trial. PMID- 15824188 TI - TP53 mutation spectrum in lung cancer is not different in women and men. AB - Whether women are more susceptible to lung cancer than men has been controversial. Several case-control studies suggested that women have greater risk of lung cancer compared with men at similar levels of cigarette smoking, whereas some large cohort studies failed to observe this association. Other studies indicated that lung cancer may have biological characteristics and mechanisms of carcinogenesis that are gender specific. Therefore, we hypothesized that women are more susceptible to the carcinogenic effects of tobacco smoke exposure, as evidenced by a higher frequency of G:C-to-T:A somatic mutations in tumors from women in comparison with men at similar levels of tobacco smoke exposure. To investigate our hypothesis, we examined the TP53 mutational spectrum in a case-only (102 women and 201 men) series study where complete smoking information was available. A similar frequency and type of somatic TP53 mutations were observed in women and men. In conclusion, our study indicates that the TP53 mutation spectrum is similar in women and men. Our results are consistent with a recent large cohort study and summary of previous cohort studies, suggesting that women likely have equivalent susceptibility to lung cancer as men. PMID- 15824189 TI - CYP2E1 G1532C, NQO1 Pro187Ser, and CYP1B1 Val432Leu polymorphisms are not associated with risk of squamous cell carcinoma of the head and neck. PMID- 15824190 TI - An inactivation stabilizer of the Na+ channel acts as an opportunistic pore blocker modulated by external Na+. AB - The Na+ channel is the primary target of anticonvulsants carbamazepine, phenytoin, and lamotrigine. These drugs modify Na+ channel gating as they have much higher binding affinity to the inactivated state than to the resting state of the channel. It has been proposed that these drugs bind to the Na+ channel pore with a common diphenyl structural motif. Diclofenac is a widely prescribed anti-inflammatory agent that has a similar diphenyl motif in its structure. In this study, we found that diclofenac modifies Na+ channel gating in a way similar to the foregoing anticonvulsants. The dissociation constants of diclofenac binding to the resting, activated, and inactivated Na+ channels are approximately 880 microM, approximately 88 microM, and approximately 7 microM, respectively. The changing affinity well depicts the gradual shaping of a use-dependent receptor along the gating process. Most interestingly, diclofenac does not show the pore-blocking effect of carbamazepine on the Na+ channel when the external solution contains 150 mM Na+, but is turned into an effective Na+ channel pore blocker if the extracellular solution contains no Na+. In contrast, internal Na+ has only negligible effect on the functional consequences of diclofenac binding. Diclofenac thus acts as an "opportunistic" pore blocker modulated by external but not internal Na+, indicating that the diclofenac binding site is located at the junction of a widened part and an acutely narrowed part of the ion conduction pathway, and faces the extracellular rather than the intracellular solution. The diclofenac binding site thus is most likely located at the external pore mouth, and undergoes delicate conformational changes modulated by external Na+ along the gating process of the Na+ channel. PMID- 15824191 TI - Conduction through the inward rectifier potassium channel, Kir2.1, is increased by negatively charged extracellular residues. AB - Ion channel conductance can be influenced by electrostatic effects originating from fixed "surface" charges that are remote from the selectivity filter. To explore whether surface charges contribute to the conductance properties of Kir2.1 channels, unitary conductance was measured in cell-attached recordings of Chinese hamster ovary (CHO) cells transfected with Kir2.1 channels over a range of K+ activities (4.6-293.5 mM) using single-channel measurements as well as nonstationary fluctuation analysis for low K+ activities. K+ ion concentrations were shown to equilibrate across the cell membrane in our studies using the voltage-sensitive dye DiBAC4(5). The dependence of gamma on the K+ activity (a(K)) was fit well by a modified Langmuir binding isotherm, with a nonzero intercept as a(K) approaches 0 mM, suggesting electrostatic surface charge effects. Following the addition of 100 mM N-methyl-D-glucamine (NMG+), a nonpermeant, nonblocking cation or following pretreatment with 50 mM trimethyloxonium (TMO), a carboxylic acid esterifying agent, the gamma-a(K) relationship did not show nonzero intercepts, suggesting the presence of surface charges formed by glutamate or aspartate residues. Consistent with surface charges in Kir2.1 channels, the rates of current decay induced by Ba2+ block were slowed with the addition of NMG or TMO. Using a molecular model of Kir2.1 channels, three candidate negatively charged residues were identified near the extracellular mouth of the pore and mutated to cysteine (E125C, D152C, and E153C). E153C channels, but not E125C or D152C channels, showed hyperbolic gamma a(K) relationships going through the origin. Moreover, the addition of MTSES to restore the negative charges in E53C channels reestablished wild-type conductance properties. Our results demonstrate that E153 contributes to the conductance properties of Kir2.1 channels by acting as a surface charge. PMID- 15824192 TI - AKT2/3 subunits render guard cell K+ channels Ca2+ sensitive. AB - Inward-rectifying K+ channels serve as a major pathway for Ca2+-sensitive K+ influx into guard cells. Arabidopsis thaliana guard cell inward-rectifying K+ channels are assembled from multiple K+ channel subunits. Following the recent isolation and characterization of an akt2/3-1 knockout mutant, we examined whether the AKT2/3 subunit carries the Ca2+ sensitivity of the guard cell inward rectifier. Quantification of RT-PCR products showed that despite the absence of AKT2 transcripts in guard cells of the knockout plant, expression levels of the other K+ channel subunits (KAT1, KAT2, AKT1, and AtKC1) remained largely unaffected. Patch-clamp experiments with guard cell protoplasts from wild type and akt2/3-1 mutant, however, revealed pronounced differences in Ca2+ sensitivity of the K+ inward rectifier. Wild-type channels were blocked by extracellular Ca2+ in a concentration- and voltage-dependent manner. Akt2/3-1 mutants lacked the voltage-dependent Ca2+ block, characteristic for the K+ inward rectifier. To confirm the akt2/3-1 phenotype, two independent knockout mutants, akt2-1 and akt2::En-1 were tested, demonstrating that the loss of AKT2/3 indeed affects the Ca2+ dependence of guard cell inward rectifier. In contrast to AKT2 knockout plants, AKT1, AtKC1, and KAT1 loss-of-function mutants retained Ca2+ block of the guard cell inward rectifier. When expressed in HEK293 cells, AKT2 channel displayed a pronounced susceptibility toward extracellular Ca2+, while the dominant guard cell K+ channel KAT2 was Ca2+ insensitive. Thus, we conclude that the AKT2/3 subunit constitutes the Ca2+ sensitivity of the guard cell K+ uptake channel. PMID- 15824193 TI - On the gating mechanisms of the light-dependent conductance in Pecten hyperpolarizing photoreceptors: does light remove inactivation in voltage dependent K channels? AB - The hyperpolarizing receptor potential of ciliary photoreceptors of scallop and other mollusks is mediated by a cGMP-activated K conductance; these cells also express a transient potassium current triggered by depolarization. During steady illumination, the outward currents elicited by voltage steps lose their decay kinetics. One interesting conjecture that has been proposed is that the currents triggered by light and by depolarization are mediated by the same population of channels, and that illumination evokes the receptor potential by removing their steady-state inactivation. Exploiting the information that has become available on the phototransduction cascade of ciliary photoreceptors, we demonstrated that the same downstream signaling elements are implicated in the modulation of voltage-elicited currents: direct chemical stimulation both at the level of the G protein and of the final messenger that controls the light-dependent channels (cGMP) also attenuate the falling phase of the voltage-activated current. Application of a protein kinase G antagonist was ineffective, suggesting that a cGMP-initiated phosphorylation step is not implicated. To ascertain the commonality of ionic pathways we used pharmacological blockers. Although millimolar 4-aminopyridine (4-AP) suppressed both currents, at micromolar concentrations only the photocurrent was blocked. Conversely, barium completely and reversibly antagonized the transient voltage-activated current with no detectable effect on the light-evoked current. These results rule out that the same ionic pores mediate both currents; the mechanism of light modulation of the depolarization-evoked K current was elucidated as a time-dependent increase in the light-sensitive conductance that is superimposed on the inactivating K current. PMID- 15824194 TI - Fructose-induced fatty liver disease: hepatic effects of blood pressure and plasma triglyceride reduction. AB - The most known risk factor for nonalcoholic fatty liver disease (NAFLD) is the metabolic syndrome. In this study, we characterized changes in liver pathology, hepatic lipid composition, and hepatic iron concentration (HIC) occurring in rats given fructose-enriched diet (FED), with and without therapeutic maneuvers to reduce blood pressure and plasma triglycerides. Rats were given FED or standard rat chow for 5 weeks. Rats on FED were divided into 4 groups: receiving amlodipine (15 mg/kg per day), captopril (90 mg/kg per day), bezafibrate (10 mg/kg per day) in the last 2 weeks, or a control group that received FED only. FED rats had hepatic macrovesicular and microvesicular fat deposits develop, with increase in hepatic triglycerides (+198%) and hepatic cholesterol (+89%), but a decrease in hepatic phospholipids (-36%), hypertriglyceridemia (+223%), and hypertension (+15%), without increase in HIC. Amlodipine reduced blood pressure ( 18%), plasma triglycerides (-12%), but there was no change in hepatic triglycerides and phospholipids concentrations. Captopril reduced blood pressure (-24%), plasma triglycerides (-36%), hepatic triglycerides (-51%), and hepatic macrovesicular fat (-51%), but increased HIC (+23%), with a borderline increase in hepatic fibrosis. Bezafibrate reduced plasma triglycerides (-49%), hepatic triglycerides (-78%), hepatic macrovesicular fat (-90%), and blood pressure ( 11%). We conclude that FED rats can be a suitable model for human NAFLD. Drugs administered to treat various aspects of the metabolic syndrome could have hepatic effects. An increase in HIC in rats with NAFLD could be associated with increased hepatic fibrosis. PMID- 15824195 TI - Mechanisms of insulin resistance in humans and possible links with inflammation. AB - Insulin resistance is a major player in the pathogenesis of the metabolic syndrome and type 2 diabetes, and yet, the mechanisms responsible for it remain poorly understood. Magnetic resonance spectroscopy studies in humans suggest that a defect in insulin-stimulated glucose transport in skeletal muscle is the primary metabolic abnormality in insulin-resistant type 2 diabetics. Fatty acids appear to cause this defect in glucose transport by inhibiting insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and IRS-1 associated phosphatidylinositol 3-kinase activity. A number of different metabolic abnormalities may increase intramyocellular/intrahepatic fatty acid metabolites; these include increased fat delivery to muscle/liver as a consequence of either excess energy intake or defects in adipocyte fat metabolism and acquired or inherited defects in mitochondrial fatty acid oxidation. Understanding the molecular/biochemical defects responsible for insulin resistance is beginning to unveil novel therapeutic targets for treatment of the metabolic syndrome and type 2 diabetes. PMID- 15824196 TI - Role of NAD(P)H oxidase- and mitochondria-derived reactive oxygen species in cardioprotection of ischemic reperfusion injury by angiotensin II. AB - Reactive oxygen species (ROS) participate in cardioprotection of ischemic reperfusion (I/R) injury via preconditioning mechanisms. Mitochondrial ROS have been shown to play a key role in this process. Angiotensin II (Ang II) exhibits pharmacological preconditioning; however, the involvement of NAD(P)H oxidase, known as an ROS-generating enzyme responsive to Ang II stimuli, in the preconditioning process remains unclear. We compared the effects of 5 hydroxydecanoate (5-HD; an inhibitor of mitochondrial ATP-sensitive potassium channels), apocynin (an NAD(P)H oxidase inhibitor), and 4-hydroxy-2,2,6,6 tetramethyl piperidinoxyl (tempol; a membrane permeable radical scavenger) on pharmacological preconditioning by Ang II in rat cardiac I/R injury in vivo. Treatment with a pressor dose of Ang II before a 30-minute coronary occlusion reduced infarct size as determined 24 hours after reperfusion. The protective effects of Ang II were eliminated by pretreatment with 5-HD or apocynin, similar to tempol. Both 5-HD and apocynin suppressed the enhanced cardiac lipid peroxidation and activation of the apoptosis signal-regulating kinase/p38, c-Jun NH2-terminal kinase (JNK) pathways, but not the Raf/MEK/extracellular signal regulated kinase pathway, elicited by acutely administered Ang II. Apocynin but not 5-HD suppressed Ang II-induced augmentations of the NAD(P)H oxidase complex formation (p47phox, p22phox, and Rac-1) and its activity in the heart. Finally, 5 HD suppressed superoxide production by isolated cardiac mitochondria without any effect on their respiration. These results suggest that the preconditioning effects of Ang II for cardiac I/R injury may be mediated by cardiac mitochondria derived ROS enhanced through NAD(P)H oxidase via JNK and p38 mitogen-activated protein kinase activation. PMID- 15824197 TI - The antioxidant edaravone attenuates pressure overload-induced left ventricular hypertrophy. AB - The free radical scavenger 3-methyl-1-phenyl-2-pyrazolin-5-one (edaravone) is used to treat patients with ischemic brain damage. We and others reported previously that in vitro and in vivo reactive oxygen species (ROS) act as second messengers to develop cardiac hypertrophy. In this study, we used an in vivo murine model of pressure overload-induced cardiac hypertrophy to examine the effects of edaravone on left ventricular hypertrophy. The animals were subjected to the transverse thoracic aorta constriction, and edaravone (10 mg/kg) was infused intraperitoneally twice daily. Seven days after the operation, we observed a significant increase in ROS production in hearts, which was eliminated by the treatment with edaravone. Pressure-overloaded hearts showed a significant increase in left ventricular weight/body weight ratio and the expression level of atrial natriuretic factor mRNA, which were attenuated by edaravone. It also reduced perivascular and intermuscular fibrosis and inhibited pressure overload induced activation of apoptosis signal-regulating kinase 1 (ASK1) and its downstream kinases of c-Jun N-terminal protein kinase and p38 mitogen-activated protein kinase. Edaravone attenuated the hypertrophic response even when the treatment was started after the onset of cardiac hypertrophic response. These findings indicate that edaravone significantly attenuates pressure overload induced cardiac hypertrophy mediated through its antioxidative function and subsequent inhibition of ASK1 signaling pathway. PMID- 15824198 TI - Diabetic brain damage in hypertension: role of renin-angiotensin system. AB - Diabetes and hypertension are potent risk factors for cerebrovascular disease. We studied the effects of an angiotensin II type 1 receptor blockade (ARB) on brain damage in hypertensives in relation to diabetes. We studied cerebral metabolism (by proton magnetic resonance spectroscopy) and hemodynamics (by phase-contrast magnetic resonance angiography) before and 3 to 4 months after candesartan therapy in 20 diabetic hypertensives (DHTs) and 20 matched nondiabetic hypertensives (HTs). Silent multiple cerebral infarcts detected by brain MRI were more common in DHTs than in HTs (50% versus 25%). Cerebral N-acetyl aspartate (NAA; an indicator of functional neuronal mass) was lower in DHTs than in HTs (8.35 versus 9.58 mmol/kg; P=0.007). Baseline quantitative volume flow in the internal carotid arteries (ICAs) and the middle cerebral arteries (MCAs) was comparable between the 2 groups, whereas cerebrovascular reserve (CVR) assessed using acetazolamide (a cerebral arteriolar dilator) in ICAs (25% versus 35%; P=0.03) and MCAs (20% versus 31%; P=0.01) was lower in DHTs than in HTs. These baseline CVR and NAA values of DHT group were lower than those of 12 matched normotensives (CVR: 44% for ICA; 41% for MCA; NAA: 10.5 mmol/kg; all P<0.005). After candesartan therapy, CVR in ICAs and MCAs was significantly increased (P=0.001) independently of the reduction of the 24-hour blood pressure level, whereas the cerebral NAA level did not change. In conclusion, brain damage is advanced in DHTs. ARB partly improved the impaired cerebral microvascular function in DHTs. PMID- 15824199 TI - Pulmonary hypertension in a GTP-cyclohydrolase 1-deficient mouse. AB - BACKGROUND: GTP-cyclohydrolase 1 (GTP-CH1) catalyzes the first step for the de novo production of tetrahydrobiopterin (BH4), a cofactor for nitric oxide synthase (NOS). The hyperphenylalaninemic mutant mouse (hph-1) displays a 90% reduction in GTP-CH1 activity. Reduced BH4 decreases NOS activity and may lead to endothelial dysfunction, and there is increasing evidence that a dysfunction of the NOS pathway may be implicated in pulmonary hypertension. The aim of the study was to investigate whether reduced BH4 in the hph-1 mouse results in a pulmonary hypertensive phenotype. METHODS AND RESULTS: Morphological characterization of the heart, lung, and kidney and measurements of systemic and right ventricular blood pressures were performed in both hph-1 and wild-type mice. BH4 and NO(x) levels were also measured. Hph-1 mice had significantly lower NO(x) and BH4 levels, consistent with previous findings. Both morphological and in vivo data were indicative of a pulmonary but not systemic hypertensive phenotype. We observed increased right ventricle-left ventricle plus septum ratios attributable only to an increase in right ventricular mass, increased smooth muscle medial area in pulmonary resistance vessels, and significantly higher right ventricular pressures in vivo. There were no significant differences between left ventricular masses and systemic pressures, and there was no observed evidence of systemic hypertension in kidney sections between wild-type and hph-1. CONCLUSIONS: This study demonstrates that mice deficient in GTP-CH1/BH4 display a pulmonary hypertensive but not systemic hypertensive phenotype. PMID- 15824200 TI - Pivotal role for endothelial tetrahydrobiopterin in pulmonary hypertension. AB - BACKGROUND: Pulmonary hypertension is a fatal disease characterized by vasoconstriction and vascular remodeling. Loss of endothelial nitric oxide bioavailability is implicated in pulmonary hypertension pathogenesis. Recent evidence suggests that the cofactor tetrahydrobiopterin (BH4) is an important regulator of nitric oxide synthase enzymatic function. METHODS AND RESULTS: In the hph-1 mouse with deficient BH4 biosynthesis, BH4 deficiency caused pulmonary hypertension, even in normoxic conditions, and greatly increased susceptibility to hypoxia-induced pulmonary hypertension. In contrast, augmented BH4 synthesis in the endothelium, by targeted transgenic overexpression of GTP-cyclohydrolase I (GCH), prevented hypoxia-induced pulmonary hypertension. Furthermore, specific augmentation of endothelial BH4 in hph-1 mice by crossing with GCH transgenic mice rescued pulmonary hypertension induced by systemic BH4 deficiency. Lung BH4 availability controlled pulmonary vascular tone, right ventricular hypertrophy, and vascular structural remodeling in a dose-dependent manner in both normoxia and hypoxia. Furthermore, BH4 availability had striking effects on the immediate vasoconstriction response to acute hypoxia. These effects of BH4 were mediated through the regulation of nitric oxide compared with superoxide synthesis by endothelial nitric oxide synthase. CONCLUSIONS: Endothelial BH4 availability is essential for maintaining pulmonary vascular homeostasis, is a critical mediator in the pathogenesis of pulmonary hypertension, and is a novel therapeutic target. PMID- 15824201 TI - Impact of high glucose/high insulin and dichloroacetate treatment on carbohydrate oxidation and functional recovery after low-flow ischemia and reperfusion in the isolated perfused rat heart. AB - BACKGROUND: It is believed that increasing cardiac glucose metabolism in the setting of ischemia and reperfusion is protective because of the resulting decrease in fatty acid oxidation, which improves cardiac efficiency and increases glucose oxidation relative to glycolysis; however, these conclusions are based primarily on studies in which glucose is the only carbohydrate provided. The goal of this study was to examine the effect of stimulating myocardial carbohydrate use either by increasing glucose and insulin levels or by using dichloroacetate on the response to ischemia and reperfusion in hearts perfused with physiological concentrations of lactate and pyruvate plus glucose and fatty acids. METHODS AND RESULTS: Metabolic fluxes were determined in hearts from male Sprague-Dawley rats perfused with 13C-labeled substrates using 13C/1H-NMR isotopomer analysis after 30 minutes of low-flow ischemia (0.3 mL/min) and 60 minutes of reperfusion. Measurements were made under control conditions: 5 mmol/L glucose, 1 mmol/L lactate, 0.1 mmol/L pyruvate, 0.3 mmol/L palmitate, and 50 microU/mL insulin plus dichloroacetate 5 mmol/L or glucose and insulin increased to 30 mmol/L and 1000 microU/mL, respectively. Dichloroacetate increased carbohydrate oxidation and the ratio of glucose oxidation to glycolysis but did not improve functional recovery or cardiac efficiency; however, elevated glucose and insulin levels improved functional recovery and cardiac efficiency but did not increase carbohydrate oxidation or the ratio of glucose oxidation to glycolysis. CONCLUSIONS: These data support the notion that increasing myocardial glucose use is beneficial in the setting of ischemia and reperfusion; however, the protective effect appears not to be mediated by shifting the balance between carbohydrate and fatty acid oxidation. PMID- 15824202 TI - Clinical and echocardiographic determinants of long-term survival after surgical myectomy in obstructive hypertrophic cardiomyopathy. AB - BACKGROUND: Surgical myectomy has been the standard treatment for patients with drug-refractory obstructive hypertrophic cardiomyopathy. The clinical and echocardiographic predictors of long-term survival and freedom from cardiovascular morbidity after myectomy have been unclear. METHODS AND RESULTS: We studied a consecutive cohort of 338 adult patients (age at operation 47+/-14 [range 18 to 77] years, 60% male) who underwent myectomy at our institution. Preoperative resting left ventricular outflow tract (LVOT) gradient was 66+/-32 mm Hg (range 5 to 158 mm Hg). Early postoperative mortality was 1.5% (5 deaths): 4 deaths occurred between 1978 and 1992, and 1 death occurred between 1993 and 2002. During long-term follow-up, 83% of patients reported an improvement to functional class I or II. The majority of patients (98%) had no resting LVOT gradient. Long-term survival was excellent, with 98+/-1% survival at 1 year, 95+/ 1% at 5 years, and 83+/-3% at 10 years after myectomy. Multivariable Cox regression analysis identified 5 predictors of overall mortality: (1) age > or =50 years at surgery (hazard ratio [HR] 2.8, 95% CI 1.5 to 5.1, P=0.001), (2) female gender (HR 2.5, 95% CI 1.5 to 4.3, P=0.0009), (3) history of preoperative atrial fibrillation (HR 2.2, 95% CI 1.2 to 4.0, P=0.008), (4) concomitant CABG (HR 3.7, 95% CI 1.7 to 8.2, P=0.001), and (5) preoperative left atrial diameter > or =46 mm (HR 2.9, 95% CI 1.6 to 5.4, P=0.0008). Significant predictors of late major cardiovascular events found on multivariable analysis were (1) female gender (HR 3.3, 95% CI 2.0 to 5.4, P<0.0001), (2) history of preoperative atrial fibrillation (HR 1.9, 95% CI 1.1 to 3.3, P=0.02), and (3) preoperative left atrial diameter > or =46 mm (HR 2.5, 95% CI 1.5 to 4.3, P=0.0008). CONCLUSIONS: Myectomy provides excellent relief for LVOT obstruction in patients with hypertrophic cardiomyopathy. Preoperative clinical and echocardiographic variables can predict long-term outcome after myectomy. PMID- 15824203 TI - Association of hemoglobin levels with clinical outcomes in acute coronary syndromes. AB - BACKGROUND: In the setting of an acute coronary syndrome (ACS), anemia has the potential to worsen myocardial ischemia; however, data relating anemia to clinical outcomes in ACS remain limited. METHODS AND RESULTS: We examined the association between baseline hemoglobin values and major adverse cardiovascular events through 30 days in 39,922 patients enrolled in clinical trials of ACS. After adjustment for differences in baseline characteristics and index hospitalization treatments, a reverse J-shaped relationship between baseline hemoglobin values and major adverse cardiovascular events was observed. In patients with ST-elevation myocardial infarction, when those with hemoglobin values between 14 and 15 g/dL were used as the reference, cardiovascular mortality increased as hemoglobin levels fell below 14 g/dL, with an adjusted OR of 1.21 (95% CI 1.12 to 1.30, P<0.001) for each 1-g/dL decrement in hemoglobin. At the other end of the range of hemoglobin, patients with hemoglobin values >17 g/dL also had excess mortality (OR 1.79, 95% CI 1.18 to 2.71, P=0.007). In patients with non-ST-elevation ACS, with those with hemoglobin 15 to 16 g/dL used as the reference, the likelihood of cardiovascular death, myocardial infarction, or recurrent ischemia increased as the hemoglobin fell below 11 g/dL, with an adjusted OR of 1.45 (95% CI 1.33 to 1.58, P<0.001) for each 1 g/dL decrement in hemoglobin. Patients with hemoglobin values >16 g/dL also had an increased rate of death or ischemic events (OR 1.31, 95% CI 1.03 to 1.66, P=0.027). CONCLUSIONS: Anemia is a powerful and independent predictor of major adverse cardiovascular events in patients across the spectrum of ACS. PMID- 15824204 TI - Images in cardiovascular medicine. Inverted left atrial appendage ligation mimicking thrombus. PMID- 15824205 TI - Letter regarding article by Wichterle et al, "prevalent low-frequency oscillation of heart rate: novel predictor of mortality after myocardial infarction". PMID- 15824206 TI - Letter regarding article by Wang et al, "nesiritide does not improve renal function in patients with chronic heart failure and worsening serum creatinine". PMID- 15824207 TI - Abstracts of the 45th Annual Conference on Cardiovascular Disease Epidemiology and Prevention in association with the Council on Nutrition, Physicial Activity and Metabolism. April 29-May 2, 2006, Washington, DC, USA. PMID- 15824208 TI - Molecular beacons illuminate subcellular events. PMID- 15824209 TI - Another lesson from the factor V Leiden mouse: thrombin generation drives arterial disease. PMID- 15824210 TI - Should treatment of sepsis include statins? PMID- 15824211 TI - Local gene transduction of cyclooxygenase-1 increases blood flow in injured atherosclerotic rabbit arteries. AB - BACKGROUND: Cyclooxygenase-1 (COX-1) is the rate-limiting component in the synthesis of prostacyclin (PGI2), an important vasodilator and antithrombotic molecule. In balloon-injured, atherosclerosis-free porcine arteries, COX-1 gene transduction increases PGI2 production, induces durable vasodilation, and reduces thrombus formation. We tested the effectiveness of COX-1 local gene transduction for the prevention of postangioplasty restenosis in atherosclerotic arteries in a hypercholesterolemic rabbit model. METHODS AND RESULTS: We injured 1 carotid artery in 43 Watanabe heritable hyperlipidemic rabbits and performed local gene transduction using a viral vector containing the COX-1 gene (AdCOX-1, n=22) or no genes (Adnull, n=21). Three days later, AdCOX-1-treated arteries stimulated with arachidonic acid produced 100% more PGI2 (P<0.01), 400% more prostaglandin E2 (PGE2) (P<0.01), 400% more prostaglandin E1 (PGE1) (P<0.01), and 250% more cAMP (P<0.05) than Adnull-treated arteries. Twenty-eight days after treatment, Doppler sonography showed that blood flow velocity was preserved in AdCOX-1-treated arteries (ratio 0.92, injured compared with contralateral uninjured carotid artery) but reduced in Adnull-treated arteries (ratio 0.39), suggesting that AdCOX-1 prevented restenosis after injury. COX-1-transduced arteries also showed 80% greater lumen area 28 days after injury (P<0.01). CONCLUSIONS: The effectiveness of COX-1 in preventing restenosis and preserving normal blood flow 28 days after injury results from increased lumen area caused by durable vasodilation. COX-1 efficacy correlates with an early increase in the production of PGI2, PGE2, PGE1 (known to cause vasodilation), and cAMP. These results demonstrate for the first time that COX-1 gene transduction is an effective treatment for the prevention of postangioplasty restenosis of atherosclerotic arteries under clinically relevant conditions. PMID- 15824212 TI - Simvastatin blunts endotoxin-induced tissue factor in vivo. AB - BACKGROUND: Beyond lipid lowering, various antiinflammatory properties have been ascribed to statins. Moreover, in vitro studies have suggested the presence of anticoagulant effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, as lipopolysaccharide (LPS)-induced monocyte tissue factor (TF) was suppressed. In this study, we examined the role of statins in experimental endotoxemia on inflammatory and procoagulant responses in vivo. METHODS AND RESULTS: In this double-blind, placebo-controlled, parallel-group study, 20 healthy, male subjects were randomized to receive either simvastatin (80 mg/d) or placebo for 4 days before intravenous administration of LPS (20 IU/kg IV). Plasma high-sensitive C-reactive protein (hsCRP), monocyte chemoattractant protein (MCP 1), sCD40L, sCD40, and prothrombin fragment F1+2 (F1.2) were determined by ELISAs at baseline and at 4 and 8 hours after LPS administration. Monocyte TF expression and monocyte-platelet aggregates were measured by whole-blood flow cytometry over the same time course. The increases in hsCRP and MCP-1, both known inducers of TF, were significantly suppressed by statin treatment after LPS challenge. Statin premedication blunted the increase of monocyte TF expression in response to LPS. In parallel, endotoxin-induced formation of F1.2 was significantly reduced by simvastatin after 4 and 8 hours. LPS infusion affected neither the formation and activation of monocyte-platelet aggregates nor plasma levels of sCD40 and sCD40L. CONCLUSIONS: Simvastatin suppresses the inflammatory response to endotoxin and blunts monocyte TF expression but does not affect platelet activation. PMID- 15824213 TI - Increasing high-density lipoprotein cholesterol in dyslipidemia by cholesteryl ester transfer protein inhibition: an update for clinicians. AB - Reduced HDL cholesterol may be a risk factor comparable in importance to increased LDL cholesterol. Interventions that raise HDL are antiatherosclerotic, presumably through acceleration of reverse cholesterol transport and by antioxidant and antiinflammatory effects. In the hypercholesterolemic rabbit, HDL levels can be increased by >50% by inhibition of cholesteryl ester transfer protein (CETP), a molecule that plays a central role in HDL metabolism. This HDL raising effect is antiatherosclerotic in moderately severe hyperlipidemia but appears to be ineffective in the presence of severe hypertriglyceridemia. In humans, mutations resulting in CETP inhibition have been associated with both reduced and increased risk of atherosclerosis. Proposed explanations for these apparently disparate observations are that the antiatherosclerotic effect of CETP inhibition varies with either the metabolic milieu or the degree of CETP inhibition. We now have pharmacological inhibitors of CETP that are capable of increasing HDL by as much as 50% to 100% in humans. The importance of this development is that reduced HDL is a risk factor independent of LDL and that these new agents alter HDL by a magnitude comparable to that of statins on LDL. Clinical trials, now beginning, will need to identify the patient subsets in which CETP inhibition may be more or less effective. PMID- 15824214 TI - Atherothrombotic disorders: new insights from hematology. PMID- 15824215 TI - Randomized trial of treatment of amblyopia in children aged 7 to 17 years. AB - OBJECTIVE: To evaluate the effectiveness of treatment of amblyopia in children aged 7 to 17 years. METHODS: At 49 clinical sites, 507 patients with amblyopic eye visual acuity ranging from 20/40 to 20/400 were provided with optimal optical correction and then randomized to a treatment group (2-6 hours per day of prescribed patching combined with near visual activities for all patients plus atropine sulfate for children aged 7 to 12 years) or an optical correction group (optical correction alone). Patients whose amblyopic eye acuity improved 10 or more letters (> or =2 lines) by 24 weeks were considered responders. RESULTS: In the 7- to 12-year-olds (n = 404), 53% of the treatment group were responders compared with 25% of the optical correction group (P<.001). In the 13- to 17-year olds (n = 103), the responder rates were 25% and 23%, respectively, overall (adjusted P = .22) but 47% and 20%, respectively, among patients not previously treated with patching and/or atropine for amblyopia (adjusted P = .03). Most patients, including responders, were left with a residual visual acuity deficit. CONCLUSIONS: Amblyopia improves with optical correction alone in about one fourth of patients aged 7 to 17 years, although most patients who are initially treated with optical correction alone will require additional treatment for amblyopia. For patients aged 7 to 12 years, prescribing 2 to 6 hours per day of patching with near visual activities and atropine can improve visual acuity even if the amblyopia has been previously treated. For patients 13 to 17 years, prescribing patching 2 to 6 hours per day with near visual activities may improve visual acuity when amblyopia has not been previously treated but appears to be of little benefit if amblyopia was previously treated with patching. We do not yet know whether visual acuity improvement will be sustained once treatment is discontinued; therefore, conclusions regarding the long-term benefit of treatment and the development of treatment recommendations for amblyopia in children 7 years and older await the results of a follow-up study we are conducting on the patients who responded to treatment. PMID- 15824216 TI - Verteporfin therapy of subfoveal minimally classic choroidal neovascularization in age-related macular degeneration: 2-year results of a randomized clinical trial. AB - OBJECTIVE: To compare the treatment effect and safety of photodynamic therapy with verteporfin using a standard (SF) or reduced (RF) light fluence rate with that of placebo therapy in patients with subfoveal minimally classic choroidal neovascularization (CNV) with age-related macular degeneration. DESIGN: Phase 2, multicenter, double-masked, placebo-controlled, randomized clinical trial. SETTING: Nineteen ophthalmology practices in North America and Europe. PARTICIPANTS: Patients with initial best-corrected visual acuity of at least 20/250 and a lesion size of no greater than 6 Macular Photocoagulation Study (MPS) disc areas. METHODS: We randomly assigned 117 patients (1:1:1) to verteporfin infusion (6 mg/m(2)) and light application with an RF rate (300 mW/cm(2)) for 83 seconds (light dose of 25 J/cm(2)) or an SF rate (600 mW/cm(2)) for 83 seconds (light dose of 50 J/cm(2)) or to placebo infusion with RF or SF. Treatment was repeated every 3 months if the treating physician noted fluorescein leakage from CNV on angiography. Patients in whom a predominantly classic lesion developed could receive open-label standard verteporfin treatment. Best-corrected visual acuity was measured every 3 months, and angiographic changes were assessed by the Photograph Reading Center through the 3-month examination unless an ocular adverse event or conversion to a predominantly classic lesion was identified by an investigator. Safety was assessed throughout the study. All outcomes were on an intent-to-treat basis. RESULTS: One hundred three (88%) of 117 patients completed the 24-month examination. Twelve (30%) of 40 patients assigned to placebo received open-label standard verteporfin treatment after confirmation of presence of predominantly classic CNV. At month 12, a loss of at least 3 lines of visual acuity occurred in 5 (14%) of 36 eyes assigned to RF and 10 (28%) of 36 eyes assigned to SF, compared with 18 (47%) of 38 eyes assigned to placebo (RF, P = .002; SF, P = .08; RF + SF, P = .004). At month 24, this loss occurred in 9 (26%) of 34 eyes assigned to RF and 17 (53%) of 32 assigned to SF, compared with 23 (62%) of 37 eyes assigned to placebo (RF, P = .003; SF, P = .45; RF + SF, P = .03). Progression to predominantly classic CNV by 24 months was more common in the placebo group (11 [28%] of 39 patients compared with 2 [5%] of 38 in the RF group [P = .007] and 1 [3%] of 37 in the SF group [P = .002]). No unexpected ocular or systemic adverse events were identified. Treatment-related, usually transient visual disturbances were 13% with SF, 10% with placebo, and 5% with RF. CONCLUSIONS: Verteporfin therapy safely reduced the risks of losing at least 15 letters (> or =3 lines) of visual acuity and progression to predominantly classic CNV for at least 2 years in individuals with subfoveal minimally classic lesions due to age-related macular degeneration measuring 6 MPS disc areas or less. Based on the overall evidence available on verteporfin therapy for these lesions, the VIM Study Group would consider recommending verteporfin therapy for relatively small minimally classic lesions similar to those enrolled in the VIM Trial. PMID- 15824217 TI - Oxidative DNA damage in the human trabecular meshwork: clinical correlation in patients with primary open-angle glaucoma. AB - OBJECTIVE: To evaluate the intensity of oxidative molecular damage and its clinical correlations: visual field damage, intraocular pressure, age, and disease duration. METHODS: DNA was extracted from human trabecular meshwork specimens collected from 17 glaucoma-affected patients using standard filtration surgery. Twenty-one specimens from healthy eyes collected for cornea transplants serve as controls. Oxidative DNA damage was evaluated by determining 8-hydroxy-2' deoxyguanosine levels. All patients underwent a Humphrey 30-2 visual field examination and diurnal tonometry before surgery. RESULTS: The mean +/- SD DNA oxidative damage was 8.51 +/- 5.44 and 1.75 +/- 1.80 8-hydroxy-2'-deoxyguanosine molecules/10(5) normal nucleotides in patients with glaucoma and controls, respectively. A statistically significant correlation was found among human trabecular meshwork DNA oxidative damage, visual field damage, and intraocular pressure. No other statistically significant correlations were found. CONCLUSIONS: Oxidative stress may represent an important pathogenetic step in primary open-angle glaucoma because it could induce human trabecular meshwork degeneration, favoring an intraocular pressure increase, thus priming the glaucoma pathogenetic cascade. PMID- 15824218 TI - Optical coherence tomography longitudinal evaluation of retinal nerve fiber layer thickness in glaucoma. AB - OBJECTIVES: To longitudinally evaluate optical coherence tomography (OCT) peripapillary retinal nerve fiber layer thickness measurements and to compare these measurements across time with clinical status and automated perimetry. METHODS: Retrospective evaluation of 64 eyes (37 patients) of glaucoma suspects or patients with glaucoma participating in a prospective longitudinal study. All participants underwent comprehensive clinical assessment, visual field (VF) testing, and OCT every 6 months. Field progression was defined as a reproducible decline of at least 2 dB in VF mean deviation from baseline. Progression of OCT was defined as reproducible mean retinal nerve fiber layer thinning of at least 20 mum. RESULTS: Each patient had a median of 5 usable OCT scans at median follow up of 4.7 years. The difference in the linear regression slopes of retinal nerve fiber layer thickness between glaucoma suspects and patients with glaucoma was nonsignificant for all variables; however, Kaplan-Meier survival curve analysis demonstrated a higher progression rate by OCT vs VF. Sixty-six percent of eyes were stable throughout follow-up, whereas 22% progressed by OCT alone, 9% by VF mean deviation alone, and 3% by VF and OCT. CONCLUSIONS: A greater likelihood of glaucomatous progression was identified by OCT vs automated perimetry. This might reflect OCT hypersensitivity or true damage identified by OCT before detection by conventional methods. PMID- 15824219 TI - Outcomes of complex retinal detachment repair using 1000- vs 5000-centistoke silicone oil. AB - OBJECTIVE: To compare anatomic and visual acuity outcomes, as well as complication rates, after retinal detachment repair using 1000- vs 5000 centistoke silicone oil. METHODS: Records of all patients who underwent retinal detachment repair with silicone oil at one institution between January 1, 1995, and December 31, 2000, were reviewed. Anatomic outcomes included retinal redetachment and macula-off retinal redetachment. Visual acuity outcomes included ambulatory vision (> or =5/200) and change in visual acuity from preoperative examination. Complications included rates of secondary intraocular pressure elevation, hypotony, corneal opacification, cataract, and oil emulsification. Outcomes were assessed at 1 week, 1 month, 3 months, 6 months, and 1 year. RESULTS: The study included 82 eyes that underwent retinal detachment repair with 1000-centistoke silicone oil and 243 eyes that underwent retinal detachment repair with 5000-centistoke silicone oil. Demographic characteristics, cause of retinal detachment, and preoperative ocular characteristics were similar in the 2 groups. There was no significant difference in the rate of retinal redetachment at each of the follow-up intervals investigated. The cumulative retinal detachment rate was also similar between the 2 groups except among trauma cases, for which 1000-centistoke silicone oil was associated with a higher cumulative redetachment rate (P<.001). There was no significant difference between the groups with respect to (1) change in visual acuity from preoperatively to 6 months postoperatively and (2) the proportion of patients who achieved ambulatory vision at each of the follow-up intervals investigated. Rates of elevated intraocular pressure, hypotony, corneal abnormality, cataract, and silicone oil emulsification were similar in the 2 groups. CONCLUSIONS: Anatomic and visual acuity outcomes, as well as complication rates, were similar in both groups; retinal reattachment and ambulatory vision were achieved in most eyes regardless of oil viscosity. PMID- 15824220 TI - Symptoms and findings predictive for the development of new retinal breaks. AB - OBJECTIVE: To validate the conclusion of our previous prospective study of 250 patients with isolated posterior vitreous detachment: follow-up visits are only necessary if patients mention symptoms of flashes in combination with multiple floaters or a curtain or cloud at the initial examination, or an increase in number of floaters after the initial examination. METHODS: Prospective study of 270 consecutive patients with symptomatic isolated posterior vitreous detachment. All patients completed a questionnaire detailing their symptoms and had a full eye examination at the initial examination and at follow-up visits. Logistic regression with backward elimination was used for statistical analysis. We also performed pooled analysis of our previous and present study data. RESULTS: New retinal breaks developed in 10 patients (3.7%). Multiple floaters, a curtain or cloud, hemorrhages (retinal or vitreous) at the initial examination, and an increase in the number of floaters after the initial examination were found to be predictive factors for the development of new retinal breaks. These factors were also the only significant predictors after pooled analysis of both studies (520 patients, 23 breaks). CONCLUSIONS: We assume we can formulate a safe policy for scheduling patients with isolated posterior vitreous detachment: only patients with multiple floaters, a curtain or cloud, or hemorrhages (retinal or vitreous) at the initial examination should be scheduled for reexamination. All other patients should return only if the number of floaters increases. PMID- 15824221 TI - Asteroid hyalosis in an autopsy population: The University of California at Los Angeles (UCLA) experience. AB - OBJECTIVES: To study the prevalence and associations of asteroid hyalosis (AH) in a series of autopsy eyes. METHODS: Retrospective review of the University of California at Los Angeles (UCLA) autopsy eye database from 1965 to 2000 yielded 10,801 patients. The patients' medical histories were reviewed for evidence of diabetes mellitus, hypertension, hyperlipidemia, alcohol abuse, hypercalcemia, hypothyroidism, and chronic renal failure. Autopsy records were searched for evidence of optic atrophy, macular degeneration, posterior vitreous detachment, atherosclerosis, and chronic renal failure. Asteroid hyalosis was diagnosed by examination of the autopsy eyes. Univariate and multivariate statistical methods were used to analyze our data. RESULTS: The prevalence of AH was 1.96% in this autopsy population. By chi(2) analysis, AH was significantly correlated with age (P<.001), male sex (P = .006), age-related macular degeneration (P = .02), hypertension (P = .03), atherosclerosis (P<.001), and posterior vitreous attachment (P<.001). After adjusting for age in a multivariate logistic regression analysis, statistical significance was found only for posterior vitreous attachment (P = .002) and male sex (P = .046). No statistically significant association was found with diabetes mellitus or alcohol abuse by univariate or multivariate analysis. Analysis of the odds ratio showed a strong age effect that increased from 5.0 (95% confidence interval, 2.2-11.3) in age group 41 to 50 years, compared with 25.4 (95% Wald confidence interval, 8.2-77.9) in the age group of patients older than 90 years. CONCLUSIONS: A unique epidemiological autopsy cohort study of AH and its systemic associations yielded a higher prevalence of AH than previous studies. Asteroid hyalosis was strongly correlated with age and inversely correlated with posterior vitreous detachment. Unlike some previous reports, we found no statistically significant correlation between AH and diabetes mellitus. PMID- 15824222 TI - Psychological disturbance in graves ophthalmopathy. AB - OBJECTIVE: To study mood disturbance in Graves ophthalmopathy. METHODS: Forty eight patients (mean age, 55 years; 40 women and 8 men) with Graves ophthalmopathy from a university-based referral center were classified into two groups, 24 with moderate/severe disease (study group) and 24 with negligible/mild disease (control group). The groups were matched with regard to demographic and medical characteristics. All participants completed a mood survey to assess differences in degree of emotional distress. MAIN OUTCOME MEASURE: The Profile of Mood States survey, a 65-item self-reported inventory designed to assess emotional distress, was the primary outcome measure. A total mood disturbance score was assigned by summing the scores derived on the 6 subscales of the survey -tension, depression, vigor, confusion, fatigue, and anger. RESULTS: Analysis of variance revealed that patients with moderate/severe Graves ophthalmopathy showed significantly greater emotional distress than patients with mild/negligible Graves ophthalmopathy on the Profile of Mood States mean total score (P<.001). Additionally, patients who had disfigurement (proptosis) as the predominant clinical feature had significantly elevated emotional distress compared with the control group (P = .01), whereas no significant difference was detected between the control group and patients with diplopia as the predominant clinical feature (P = .20). CONCLUSION: Patients with moderate to severe Graves ophthalmopathy have significant mood disturbance, especially when disfiguring signs are predominant. We propose that evaluation of the psychological burden of the disease should be considered in routine follow-up and in decisions regarding treatment. PMID- 15824223 TI - Reasons for delay of surgical intervention in adult strabismus. AB - BACKGROUND: It has been our impression that adult patients with strabismus frequently delay surgical intervention. OBJECTIVES: To determine the length of time adult patients waited before undergoing strabismus surgery and to determine the reasons why these delays occurred. DESIGN, SETTING, AND PARTICIPANTS: Prospective survey of consecutive patients who delayed strabismus surgery for more than 1 year. INTERVENTION: Preoperative survey. MAIN OUTCOME MEASURES: The primary outcome measure was time between the onset of strabismus and surgery and the reason why surgery had not been previously sought. RESULTS: The mean age among 128 study participants was 45.7 years (age range, 18-86 years). The mean time between the onset of current strabismus and surgery was 19.9 years (range, 1 72 years). The major reasons for delay in seeking surgical treatment included the following: surgery was never offered by eye care specialist (35 patients [27%]), surgery was offered but declined by the patient (29 patients [23%]), the patient had received prior satisfactory nonsurgical care (17 patients [13%]), the patient had never sought care (14 patients [11%]), the patient had a previous poor surgical experience (8 patients [6%]), and the patient had been told by their eye specialist that nothing could be done or that surgery could make them worse (8 patients [6%]). CONCLUSIONS: Strabismus surgery is often delayed for many years in adult patients who could potentially benefit from it. Almost half of such delays could be avoided by better education of the lay public and the medical community. PMID- 15824224 TI - Postsurgical assessment and long-term safety of recombinant adeno-associated virus-mediated gene transfer into the retinas of dogs and primates. AB - OBJECTIVE: To evaluate, in dogs and primates, the short-term effects of subretinal injection and the safety of long-term recombinant adeno-associated virus (rAAV)-mediated transgene expression with respect to retinal morphology and function. METHODS: Subretinal delivery of rAAV (serotype 2, 4, or 5) was performed unilaterally in 14 beagles and 9 macaques. Postsurgical condition was evaluated during a 2-month follow-up study. Three dogs and 1 primate were examined for the long-term study. Green fluorescent protein expression was monitored by fluorescent retinal imaging. Retinal anatomy and function were assessed by angiography and electroretinography, respectively. RESULTS: Transgene expression was observed in 20 of 23 subretinally injected animals (both with and without vitrectomy). We did not detect an inflammatory response in any of the 23 treated subjects. In the long-term study, transgene expression was detected at the latest points evaluated: 36 months for the rAAV-2-injected dog, 24 months for the rAAV-4 and rAAV-5 dogs, and more than 18 months for the rAAV-4-injected primate. Angiography examinations were performed and showed no retinal abnormalities. Functional evaluation showed normal electroretinographic amplitude responses that were similar to those of the noninjected contralateral eyes. CONCLUSIONS: Subretinal injection of the rAAV vector in dogs and primates is a safe procedure with no perioperative complications and a high rate of successful retinal gene transfer. The retinal anatomy and function remained unchanged, despite persistent transgene expression up to 36 months postinjection with rAAV 2, -4, or -5. Additionally, we observed no other adverse effects, such as tumor formation due to possible insertional mutagenesis. These short- and long-term studies on rAAV transgene expression using large animals are encouraging for the prospects of ocular gene therapy applications in humans. CLINICAL RELEVANCE: These short- and long-term studies on rAAV transgene expression using large animals are encouraging for the prospects of ocular gene therapy applications in humans. PMID- 15824225 TI - Safety and efficacy of intravitreal injection of ranibizumab in combination with verteporfin PDT on experimental choroidal neovascularization in the monkey. AB - OBJECTIVE: To study the safety and efficacy of intravitreal injections of anti vascular endothelial growth factor antibody fragment (ranibizumab [formerly known as rhuFabV2], Lucentis; Genentech, South San Francisco, Calif) in combination with intravenous verteporfin (Visudyne; Novartis, East Hanover, NJ) photodynamic therapy (PDT) on experimental choroidal neovascularization in the monkey eye. METHODS: Choroidal neovascularization was induced by laser injury in both eyes of cynomolgus monkeys and followed with weekly fundus photography and fluorescein angiography. Two weeks after induction, weekly treatments were initiated. These treatments included using either an intravitreal injection of ranibizumab (previously known as rhuFabV2) in combination with verteporfin PDT or a ranibizumab vehicle (placebo) in combination with verteporfin PDT (PDT only). Six animals (group 1) initially received intravitreal injections followed 1 week later by PDT. Four animals (group 2) initially received PDT followed 1 week later by intravitreal injection. Two animals (group 3) received injections and PDT on the same day at 2-week intervals. Photodynamic therapy was applied in all 3 groups every 2 weeks for 3 treatments with follow-up through 2 weeks after the last PDT treatment. Fluorescein angiograms were graded using a masked standardized protocol. The data were analyzed using the McNemar chi(2) test for matched pairs. RESULTS: No choroidal neovascularization leakage was observed in the eyes of animals treated with ranibizumab and PDT at day 21 or 42 after the start of the first treatment. Leakage persisted in eyes treated with PDT alone at 21 days (3 of 12 eyes) and 42 days (2 of 12 eyes). At all time points studied, the ranibizumab and PDT-treated eyes experienced better angiographic outcomes than the eyes receiving PDT alone. CONCLUSION: These preliminary data indicate that an intravitreal ranibizumab injection in combination with verteporfin PDT (ranibizumab and PDT) causes a greater reduction in angiographic leakage than PDT and intravitreal vehicle injection (PDT only) in experimental choroidal neovascularization. CLINICAL RELEVANCE: This combination therapy can potentially offer a new treatment modality for choroidal neovascularization in patients with macular degeneration and other diseases. PMID- 15824226 TI - Long-term nutrient intake and 5-year change in nuclear lens opacities. AB - OBJECTIVE: To determine if usual nutrient intake is related to a 5-year change in the amount of lens nuclear opacification assessed by computer-assisted image analysis. DESIGN: A sample of 408 Boston, Mass-area women from the Nurses' Health Study aged 52 to 74 years at baseline participated in a 5-year study related to nutrition and vision. Usual nutrient intake was calculated as the average intake from 5 food frequency questionnaires that were collected over a 13- to 15-year period before the baseline evaluation of lens nuclear density. Duration of vitamin supplement use before baseline was determined from 7 questionnaires collected during this same period. We assessed the degree of nuclear density (opacification) using computer-assisted image analysis of digital lens images with amount of nuclear density measured as a function of average pixel gray scale, ranging from 0 (clear) to 255 (black). RESULTS: Median (range) baseline and follow-up nuclear densities were 44 (19 to 102) and 63 (32 to 213). The median (range) 5-year change in nuclear density was 18 (-29 to 134) and was positively correlated with the amount of opacification at baseline (Spearman correlation coefficient = 0.35; P<.001). Geometric mean 5-year change in nuclear density was inversely associated with the intake of riboflavin (P trend = .03) and thiamin (P trend = .04) and duration of vitamin E supplement use (P trend = .006). CONCLUSION: Our results suggest that long-term use of vitamin E supplements and higher riboflavin and/or thiamin intake may reduce the progression of age-related lens opacification. PMID- 15824227 TI - Anterior chamber depth and the risk of primary angle closure in 2 East Asian populations. AB - OBJECTIVE: To evaluate the risk of peripheral anterior synechiae (PAS) and glaucomatous optic neuropathy attributable to primary angle closure at varying anterior chamber depths (ACDs) in two East Asian populations. METHODS: Participants 40 years and older were from 2 population-based glaucoma surveys in Mongolia and Singapore. Central ACD was measured by optical pachymetry. Presence of PAS was determined by dynamic gonioscopy. Cases with secondary PAS were excluded. Glaucomatous optic neuropathy was diagnosed in subjects with structural and functional evidence of glaucoma. RESULTS: A total of 2032 subjects, consisting of 942 Mongolians and 1090 Chinese Singaporeans, were included in this study. A logistic model of the relationship between ACD and PAS among Singaporeans showed a consistent, incremental increase in PAS across the entire range of ACD. In deeper anterior chambers the rate of PAS exceeded that seen in Mongolians. In Mongolians, there was a clear threshold for ACD (2.4 mm) at or above which PAS were very uncommon. With ACD less than 2.4 mm, the rate of PAS rose rapidly to overtake that seen in Singaporeans. CONCLUSIONS: Shallow anterior chambers are a significant risk factor for angle closure in East Asians, although the nature of the association is specific to the individual population. There was a trend toward higher rates of glaucomatous optic neuropathy in people with the shallowest anterior chambers. PMID- 15824228 TI - Patient expectations regarding eye care: development and results of the Eye Care Expectations Survey (ECES). AB - OBJECTIVE: To develop an instrument (the Eye Care Expectations Survey [ECES]) that can identify and quantify the expectations of patients visiting eye care providers. METHODS: A total of 202 patients attending 4 ophthalmology practices affiliated with Duke University Eye Center enrolled in the study. All participants completed the pilot version of the ECES, which was developed through a review of the expectations literature and a multicondition focus group process. Factor analysis of patients' responses was used to identify the performance characteristics of the ECES. RESULTS: Factor analysis yielded factors that describe 4 distinct types of expectations: patient involvement in eye care, interpersonal manner, information about diagnosis and prognosis, and communication and clinical competence. These 4 factors explained 89% of the total variance. The factor analysis identified a subset of 27 questions (of the original 37) to represent the 4 factors. Estimates of internal consistency and test-retest reproducibility indicate that the ECES is reliable. In addition, tests of association with clinical variables support the construct validity of the survey. The psychometric properties of the ECES were influenced by the severity of the underlying eye condition as well as other demographic and clinical variables, including the patient's rating of his or her own vision, best corrected visual acuity, sex, education, race, and household income. CONCLUSIONS: Based on this cross-sectional study, the 27-item ECES appears to be a useful tool for better understanding patients' expectations regarding eye care. PMID- 15824229 TI - A novel TIMP3 mutation associated with Sorsby fundus dystrophy. PMID- 15824230 TI - Henry "Jullundur" Smith's "Extraction of cataract in the capsule": a landmark article. PMID- 15824231 TI - A blue-blocking intraocular lens should be used in cataract surgery. PMID- 15824232 TI - Intraocular lenses should block UV radiation and violet but not blue light. PMID- 15824233 TI - Treatment of amblyopia in older children. PMID- 15824234 TI - Oklahoma's optometrists encroach on surgical rights of ophthalmologists; place politics above patient safety. PMID- 15824235 TI - Sebaceous adenoma of the eyelid in Muir-Torre syndrome. PMID- 15824236 TI - Desmoplastic small round cell tumor: a potentially lethal neoplasm manifesting in the orbit with associated visual symptoms. PMID- 15824237 TI - Optical coherence tomography of optic disc swelling in acute primary angle closure glaucoma. PMID- 15824238 TI - Cavitary melanoma of ciliary body simulating a cyst. PMID- 15824239 TI - Arteriovenous dissection in a living human eye: clinicopathologic correlation. PMID- 15824240 TI - Cytomegalovirus retinitis in an immunocompetent patient. PMID- 15824241 TI - Chronic myelogenous leukemia manifested as bilateral proliferative retinopathy. PMID- 15824242 TI - Clinical comparison of the proview eye pressure monitor with the Goldmann Applanation Tonometer and the TonoPen. PMID- 15824244 TI - The utility of 0.5% apraclonidine in the diagnosis of horner syndrome. PMID- 15824246 TI - Optical coherence tomography and an intraretinal lesion. PMID- 15824247 TI - Prevalence of refractive errors among adults in Europe. PMID- 15824248 TI - It's time for a clinical trial to investigate intravitreal triamcinolone for macular edema due to retinal vein occlusion: the SCORE study. PMID- 15824249 TI - Subthalamic nucleus vs globus pallidus interna deep brain stimulation, the rematch: will pallidal deep brain stimulation make a triumphant return? PMID- 15824250 TI - Management of acute cerebellar stroke. AB - Acute cerebellar infarction or hemorrhage may initially manifest in a clinically indolent manner only to later deteriorate into a life-threatening neurologic catastrophe. At the other end of the spectrum, some patients with cerebellar stroke may present in a moribund comatose state. In both patient groups, it is often unclear at what point surgical intervention should be considered either to prevent further neurologic deterioration or to try to salvage a meaningful neurologic recovery. In this review, we present clinical cases that illustrate decision points in the management of patients with acute cerebellar stroke, with emphasis on clinical and imaging characteristics. We conclude with an analysis of clinical decision making in the management of patients with space-occupying cerebellar stroke. The management of acute cerebellar infarction or hemorrhage often requires difficult and prompt decisions by treating neurologists, and certain easily identifiable clinical and imaging findings may assist in appropriate patient triage and timely neurosurgical intervention. PMID- 15824251 TI - Human prion diseases: molecular and clinical aspects. AB - Compared with that of other human pathogens, the proposed replicative cycle of prions is disarmingly simple. It encompasses misfolding of a single protein, the cellular prion protein (PrPC), into a disease-associated form called PrPSc. This is followed by PrPSc aggregation and possibly fragmentation of aggregates, which may augment the number of replicative units. Although there is no formal proof of the correctness of this model, a wealth of evidence indicates that pathogen encoded informational nucleic acids are dispensable for prion replication. Despite the simplicity of the replicative process, the human phenotypic range of prion diseases is extremely variable and includes the sporadic, inherited, and acquired forms of Creutzfeldt-Jakob disease. In addition, prion diseases occur in a wide range of animals and can be propagated within and between animal species. The present review article discusses current concepts and controversies surrounding the basic biological features of prions. PMID- 15824252 TI - Pallidal vs subthalamic nucleus deep brain stimulation in Parkinson disease. AB - BACKGROUND: Deep brain stimulation (DBS) of the globus pallidus interna (GPi) and subthalamic nucleus (STN) has been reported to relieve motor symptoms and levodopa-induced dyskinesia in patients with advanced Parkinson disease (PD). Although it has been suggested that stimulation of the STN may be superior to stimulation of the GPi, comparative trials are limited. OBJECTIVE: To extend our randomized, blinded pilot comparison of the safety and efficacy of STN and GPi stimulation in patients with advanced PD. DESIGN: This study represents the combined results from our previously published, randomized, blinded, parallel group pilot study and additional patients enrolled in our single-center extension study. SETTING: Oregon Health and Science University in Portland.Patients Twenty three patients with idiopathic PD, levodopa-induced dyskinesia, and response fluctuations were randomized to implantation of bilateral GPi or STN stimulators. Patients and evaluating clinicians were blinded to stimulation site. All patients were tested preoperatively while taking and not taking medications and after 3, 6, and 12 months of DBS. MAIN OUTCOME MEASURES: Postoperatively, response of symptoms to DBS, medication, and combined medication and DBS was evaluated. Twenty patients (10 in the GPi group and 10 in the STN group) completed 12-month follow-up. RESULTS: Off-medication Unified Parkinson's Disease Rating Scale motor scores were improved after 12 months of both GPi and STN stimulation (39% vs 48%). Bradykinesia tended to improve more with STN than GPi stimulation. No improvement in on-medication function was observed in either group. Levodopa dose was reduced by 38% in STN stimulation patients compared with 3% in GPi stimulation patients (P = .08). Dyskinesia was reduced by stimulation at both GPi and STN (89% vs 62%). Cognitive and behavioral complications were observed only in combination with STN stimulation. CONCLUSION: Stimulation of either the GPi or STN improves many features of advanced PD. It is premature to exclude GPi as an appropriate target for DBS in patients with advanced disease. PMID- 15824253 TI - Interferon beta promotes nerve growth factor secretion early in the course of multiple sclerosis. AB - BACKGROUND: Interferon beta therapy has been shown to reduce the rate of clinical relapse and the frequency of magnetic resonance imaging-defined T2- weighted lesions in patients with multiple sclerosis (MS). When given early, interferon beta also reduces the rate of development of brain atrophy and improves axonal integrity. Nerve growth factor (NGF) can retard the severity and course of experimental allergic encephalomyelitis. OBJECTIVE: To determine whether interferon beta effects on patients with MS could be related to modulation of neurotrophin production within the central nervous system. DESIGN: We studied neurotrophin production by human glial and brain endothelial cells in response to coculture with MS patient-derived lymphocytes, and correlated levels of NGF secretion with clinical and magnetic resonance imaging-defined markers of disease. RESULTS: We demonstrate that production of NGF by human brain microvascular endothelial cells is triggered by interaction with T lymphocytes derived from MS patients. No such response was observed using human adult microglia or human fetal astrocytes. Nerve growth factor production by endothelial cells was potentiated by pretreating lymphocytes with interferon beta in vitro, and by using lymphocytes derived from MS patients treated with interferon beta in vivo. By using this assay, we show that levels of NGF induced by lymphocytes from MS patients inversely correlate with magnetic resonance imaging measures of brain atrophy and axonal injury. CONCLUSION: These findings suggest that interferon beta-mediated production of NGF at the level of the blood brain barrier, whether acting as an immunomodulator or directly on neural cells, is another potential mechanism contributing to the magnetic resonance imaging defined effect of interferon beta on brain atrophy when given early in the course of MS. PMID- 15824254 TI - Metabolite changes in normal-appearing gray and white matter are linked with disability in early primary progressive multiple sclerosis. AB - BACKGROUND: Abnormalities in normal-appearing brain tissues may contribute to disability in primary progressive multiple sclerosis (PPMS), where few lesions are seen on conventional imaging. OBJECTIVES: To evaluate the mechanisms underlying disease progression in the early phase of PPMS by measuring metabolite concentrations in normal-appearing white matter (NAWM) and cortical gray matter (CGM) and to assess their relationship with clinical outcomes. DESIGN: Case control study. SETTING: Tertiary referral hospital. Patients Forty-three consecutive patients within 5 years of onset of PPMS and 44 healthy control subjects. MAIN OUTCOME MEASURES: Concentrations of choline-containing compounds, phosphocreatine, myo-inositol, total N-acetyl-aspartate (tNAA), and glutamate glutamine were estimated using proton magnetic resonance spectroscopic imaging. Brain parenchymal, white matter and gray matter fractions and proton density and gadolinium-enhancing lesion loads were calculated. The Expanded Disability Status Scale and Multiple Sclerosis Functional Composite scores were recorded. RESULTS: In CGM, concentrations of the tNAA (P<.001) and glutamate-glutamine (P = .005) were lower in patients with PPMS than in controls. In NAWM, myo-inositol levels were higher (P = .002) and tNAA levels were lower (P = .005) in patients with PPMS than in controls. The Expanded Disability Status Scale score correlated with the tNAA concentration in CGM (r = -0.44; P = .03) and with myo-inositol (r = 0.41; P = .01) and glutamate-glutamine concentrations (r = 0.41; P = .01) in NAWM. Proton density lesion load correlated negatively with CGM tNAA concentration and positively with NAWM myo-inositol concentration. CONCLUSION: Metabolite changes, which differ in CGM and NAWM, occur in early PPMS and are linked with disability. PMID- 15824255 TI - Early differential diagnosis of multiple sclerosis using a new oligoclonal band test. AB - BACKGROUND: Intrathecal IgG synthesis (ITGS), in conjunction with magnetic resonance imaging, can help in the early diagnosis of multiple sclerosis (MS). Recently, we developed a new oligoclonal IgG band (OCGB) test for ITGS detection that is more sensitive and easier to interpret than previously described methods. OBJECTIVE: To assess the accuracy of a new OCGB detection test in the diagnosis of MS. DESIGN: Prospective observational study. SETTING: A hospital neurology department. Patients A total of 385 patients with various neurologic disorders. MAIN OUTCOME MEASURES: The sensitivity and specificity of the OCGB detection test for MS diagnosis. RESULTS: Intrathecal IgG synthesis was found in 127 patients with MS (96.2%), 18 (35.3%) with central nervous system infections, and 1 with motor neuron disease. Two patterns reflected ITGS. One pattern, showing OCGBs restricted to cerebrospinal fluid, was predominantly found in MS. The other pattern, with OCGBs in serum and additional bands in cerebrospinal fluid, was mostly found in central nervous system infections. No patients with other inflammatory neurologic diseases showed ITGS. These patients frequently displayed a mirror pattern, with identical bands in serum and cerebrospinal fluid. Considering all patients, the sensitivity for the diagnosis of MS was 96.2%, and the specificity was 92.5%. Excluding infections, which usually do not present a differential diagnosis problem with MS, the sensitivity was still 96.2%, and the specificity increased to 99.5%. CONCLUSION: The accuracy of this OCGB method reinforces the value of cerebrospinal fluid studies in the early differential diagnosis of MS. PMID- 15824256 TI - Progressive gray matter damage in patients with relapsing-remitting multiple sclerosis: a longitudinal diffusion tensor magnetic resonance imaging study. AB - BACKGROUND: Diffusion tensor magnetic resonance imaging (DT MRI) has the potential to provide in vivo information about tissue microstructure. In multiple sclerosis (MS), DT MRI has disclosed the presence of occult structural damage in the normal-appearing brain tissues. OBJECTIVE: To investigate whether DT MRI is sensitive to longitudinal changes of brain damage that may occur beyond the resolution of T2-weighted images in patients with relapsing-remitting MS. DESIGN: Twenty-six untreated patients with relapsing-remitting MS were followed up for 18 months. Dual-echo, DT and postcontrast T1-weighted MRIs of the brain were obtained at baseline and then every 3 months. Mean diffusivity (D) histograms of normal-appearing gray (GM) and white matter were produced. Total T2-hyperintense and T1-hypointense lesion volumes; normalized whole brain tissue, GM, and white matter volumes; percentage brain volume change between the study entry and exit images; average lesion D; and fractional anisotropy were also calculated. RESULTS: During the study period, a significant decrease of normalized whole brain tissue, average lesion fractional anisotropy and normal-appearing GM D histogram peak height, and a significant increase of average normal-appearing GM D and T2-hyperintense lesion volumes were observed. Changes of normal-appearing GM diffusivity were independent of the concomitant changes of normalized whole brain tissue and GM volumes. CONCLUSIONS: The DT MRI findings show progressive microstructural changes in the normal-appearing GM of patients with untreated relapsing-remitting MS. Such changes do not reflect a concomitant development of brain atrophy and confirm the importance of GM pathology in MS. PMID- 15824257 TI - Initial evaluations for multiple sclerosis in a university multiple sclerosis center: outcomes and role of magnetic resonance imaging in referral. AB - OBJECTIVE: To evaluate diagnostic outcomes, especially as they relate to reason for referral, of patients referred to a university-based multiple sclerosis (MS) center for possible MS. METHODS: Retrospective medical record review of all new patient visits to University of Colorado Multiple Sclerosis Center, Denver, from January 1, 2001, to June 30, 2003. RESULTS: Of 281 patients referred to evaluate the possibility of MS, after initial review 33% were diagnosed with MS or possible MS by the McDonald criteria. The rest had other neurological conditions (31.5%), probable psychiatric diagnoses (22.5%), or no clear diagnosis was made (12.5%). Of patients with typical, possible, or atypical demyelinating syndromes, 71%, 27%, and 0%, respectively (P<.001), had MS or possible MS. Of the 63% of patients referred on the basis of clinical symptoms and signs, 46% were diagnosed with MS or possible MS vs 11% of patients referred primarily on the basis of abnormal brain magnetic resonance imaging (MRI) results (P<.001). Of patients referred because of abnormal MRI results who did not have MS or possible MS, 70% had a clear alternative etiology for the abnormal MRI results, including migraine, age older than 50 years, other neurological disease, or hypertension. CONCLUSIONS: A significant percentage of patients referred to a university-based MS center have little or no likelihood of having MS, and many have undiagnosed, untreated psychiatric illness or common conditions with abnormal brain MRI results. With respect to the diagnosis of MS, greater training of primary care professionals, neurologists, and radiologists is necessary. PMID- 15824258 TI - Restless legs syndrome: confirmation of linkage to chromosome 12q, genetic heterogeneity, and evidence of complexity. AB - BACKGROUND: Genes are involved in the etiology of restless legs syndrome, a common sensorimotor disorder. OBJECTIVES: To replicate and to further characterize our previously reported chromosome 12q linkage results. DESIGN: Family linkage study. SETTING AND PARTICIPANTS: A total of 276 individuals from 19 families have been examined using a selection of markers spanning the identified candidate interval on chromosome 12q. RESULTS: Two-point analyses of individual pedigrees indicated that 5 kindreds were consistent with linkage to chromosome 12q. When considering these 5 pedigrees along with the family in which linkage was originally reported, we observed a maximum 2-point logarithm-of-odds score of 5.67 (at theta = 0.10; for marker D12S1636; autosomal recessive) and a maximum multipoint logarithm-of-odds score of 8.84 between the interval defined by markers D12S326 and D12S304. Furthermore, our results also suggest the presence of heterogeneity in restless legs syndrome as linkage was formally excluded across the region in 6 pedigrees. Interestingly, significantly higher periodic leg movements during sleep indices were observed for all probands with restless legs syndrome from linked families. CONCLUSIONS: These results support the presence of a major restless legs syndrome-susceptibility locus on chromosome 12q, which has been designated as RLS1, and also suggest that at least one additional locus may be involved in the origin of this prevalent condition. PMID- 15824259 TI - Presence of alanine-to-valine substitutions in myofibrillogenesis regulator 1 in paroxysmal nonkinesigenic dyskinesia: confirmation in 2 kindreds. AB - BACKGROUND: Paroxysmal nonkinesigenic dyskinesia (PNKD) is a rare disorder characterized by attacks of involuntary movements brought on by stress, alcohol, or caffeine, but not by movement. An autosomal dominant form of this disorder was mapped to chromosome 2q33-36, and different missense mutations in exon 1 of the myofibrillogenesis regulator 1 (MR1) gene were identified recently in 2 kindreds. OBJECTIVES: To describe studies on a new pedigree with PNKD, to explore the possibility of locus heterogeneity, and to further delineate the spectrum of mutations in MR1 in 2 families with PNKD. DESIGN, SETTING, AND PATIENTS: All 10 exons of MR1 were sequenced in DNA from members of 2 pedigrees with autosomal dominant PNKD. RESULTS: Different missense mutations in exon 1 of MR1 that cosegregate with disease were identified in each multiplex family. These single nucleotide mutations predicted substitution of valine for alanine in residue 7 in one family and residue 9 in the other. The same mutations were found in the only 2 families previously published. Family history and haplotype analysis make it unlikely that the families with the same mutations are related. CONCLUSIONS: The function of MR1 is unknown, but the 2 mutations identified in the 4 families with PNKD studied to date are predicted to disrupt the amino terminal alpha-helix suggesting that this region of the gene is critical for proper gene function under stressful conditions. Study of additional families will be important to determine whether analysis of a single exon (MR1 exon 1) is sufficient for genetic testing purposes. PMID- 15824260 TI - Sex differences in clinical and genetic determinants of levodopa peak-dose dyskinesias in Parkinson disease: an exploratory study. AB - BACKGROUND: Several factors, both clinical and genetic, may account for the risk of developing levodopa-induced peak-dose dyskinesias (PDD) in patients with Parkinson disease, but it is unclear how these factors interact for modulating the individual susceptibility for PDD. OBJECTIVE: To examine clinical and genetic risk factors for determining individual susceptibility of PDD in patients with Parkinson disease. DESIGN: Cohort study. SETTING: Referral center for Parkinson disease in Calabria, southern Italy. Patients Two hundred fifty patients with Parkinson disease were screened for the presence or absence of PDD following a short-term levodopa administration, and 215 subjects were available for further evaluations, including genotypic analysis of the CA dinucleotide short tandem repeat (CAn-STR) polymorphism located in the dopamine receptor D2 gene (DRD2). RESULTS: One hundred five patients (48.8%) exhibited PDD following short-term levodopa administration, and 110 patients (51.2%) did not. Multivariate logistic regression analysis showed that independent predictors for the occurrence of PDD were female sex, earlier age at onset of Parkinson disease, longer duration of treatment, and higher dose of levodopa. Genetic factors related to the DRD2 CAn STR polymorphism were not independent predictors for PDD in the total population, but they had a strong protective effect on the appearance of PDD when the multivariate analysis was performed in men (odds ratio, 0.34 [95% confidence interval, 0.14-0.84]). In women, a genetic protective effect on PDD was not evident. CONCLUSIONS: Risk factors for PDD, both clinical and genetic, act in different ways for men and women. Genetic factors related to the DRD2 polymorphic status have a protective effect on PDD development in men but not in women. A female sex-related effect for the risk of PDD may be so strong that it overcomes any protective effect due to genetic factors. PMID- 15824261 TI - Early clinical heterogeneity in choreoacanthocytosis. AB - BACKGROUND: Choreoacanthocytosis (CHAC) is a slowly progressive multisystem disorder with involuntary movements, cognitive decline, behavioral changes, seizures, and polyneuropathy caused by mutations in the VPS13A gene. OBJECTIVE: To describe the early clinical features and possible genotype-phenotype correlation in CHAC. DESIGN AND SETTING: Case series in a tertiary care center. PATIENTS AND MAIN OUTCOME METHODS: Choreoacanthocytosis was diagnosed in 3 patients of Jewish origin from 3 unrelated families. We reviewed their medical histories and performed molecular analysis by screening all 73 exons of VPS13A. RESULTS: Trichotillomania, hypertrophic cardiomyopathy, and idiopathic hyperCKemia, in 1 patient each, preceded the development of the full clinical spectrum of CHAC by 2 to 20 years. At diagnosis, 2 patients manifested signs of overt neuromuscular involvement and were homozygous for the 6059delC mutation, whereas 1 patient had only hyporeflexia and was homozygous for the EX23del mutation. Because only 1 of the 2 patients with 6059delC had cardiomyopathy, its relevance to CHAC is unclear. CONCLUSIONS: These findings extend the knowledge of significant early clinical heterogeneity in CHAC and suggest a possible genotype phenotype correlation. Awareness of the early manifestations may prevent misdiagnosis and enable appropriate genetic counseling. PMID- 15824262 TI - Cerebellar ataxia and central nervous system whipple disease. AB - BACKGROUND: Whipple disease (WD) is an infectious disease, which may affect the central nervous system. Central nervous system symptoms are eventually present in as many as 43% of the cases. To our knowledge, cerebellar ataxia in WD has never been formally studied in any large series. OBJECTIVE: To determine the prevalence of cerebellar ataxia in central nervous system WD. RESULTS: Between January 1974 and December 2003, we identified 11 patients who met criteria for definite central nervous system WD, the second largest series to date. Surprisingly, while oculomasticatory myorrhythmia was recorded in only 1 patient (9%), cerebellar ataxia had been documented in 5 cases (45%). CONCLUSION: Our data suggest that cerebellar ataxia should be considered a more common feature of central nervous system WD. PMID- 15824263 TI - Antioxidant treatment of patients with Friedreich ataxia: four-year follow-up. AB - BACKGROUND: Decreased mitochondrial respiratory chain function and increased oxidative stress have been implicated in the pathogenesis of Friedreich ataxia (FRDA), raising the possibility that energy enhancement and antioxidant therapies may be an effective treatment. OBJECTIVE: To evaluate the long-term efficacy of a combined antioxidant and mitochondrial enhancement therapy on the bioenergetics and clinical course of FRDA. DESIGN: Open-labeled pilot trial over 47 months.Patients Seventy-seven patients with clinical and genetically defined FRDA. Intervention A combined coenzyme Q(10) (400 mg/d) and vitamin E (2100 IU/d) therapy of 10 patients with FRDA over 47 months. MAIN OUTCOME MEASURES: Clinical assessment using echocardiography and the International Cooperative Ataxia Rating Scale and cardiac and skeletal muscle bioenergetics as assessed using phosphorus P 31 magnetic resonance spectroscopy. RESULTS: There was a significant improvement in cardiac and skeletal muscle bioenergetics that was maintained throughout the 47 months of therapy. Echocardiographic data revealed significantly increased fractional shortening at the 35- and 47-month time points. Comparison with cross-sectional data from 77 patients with FRDA indicated the changes in total International Cooperative Ataxia Rating Scale and kinetic scores over the trial period were better than predicted for 7 patients, but the posture and gait and hand dexterity scores progressed as predicted. CONCLUSION: This therapy resulted in sustained improvement in mitochondrial energy synthesis that was associated with a slowing of the progression of certain clinical features and a significant improvement in cardiac function. PMID- 15824264 TI - Autonomic dysfunction in Machado-Joseph disease. AB - OBJECTIVE: Machado-Joseph disease is an autosomal dominant spinocerebellar ataxia with expanded trinucleotide repeats. Although autonomic nervous system degeneration was documented in postmortem reports, the autonomic dysfunction in patients with Machado-Joseph disease, either in clinical analysis or electrophysiological investigations, has not yet been studied in detail. METHODS: Fifteen patients with genetically confirmed Machado-Joseph disease and 34 healthy subjects were studied. The study design included a detailed questionnaire, R-R interval variation on deep breathing or Valsalva maneuver, and sympathetic skin response evoked by electrical stimulation of the median nerve or magnetic stimulation of the neck. RESULTS: Sixty-seven percent of patients had at least 3 symptoms involving different aspects of autonomic functions. Voiding problems and thermoregulatory disturbance were the most common symptoms. Ten (71%) of 14 patients had abnormal R-R interval variation with a significant reduction of the mean ratio. Prolonged latency or absence of sympathetic skin response to electrical stimulation was identified in 73% of patients and to magnetic stimulation, in 53%. R-R interval variation and sympathetic skin response showed good correlation with the functional stage. Electrical stimulation revealed the highest sensitivity, specificity, and positive predictive value compared with other tests. CONCLUSION: The present investigation documents that autonomic dysfunction is not uncommon in patients with Machado-Joseph disease and might be related to the clinical progression. PMID- 15824265 TI - Founder haplotype for Machado-Joseph disease in the Indian population: novel insights from history and polymorphism studies. AB - BACKGROUND: The ACA haplotype is associated with 72% of the expanded repeats in Machado-Joseph disease (MJD) worldwide and has been traced to a Portuguese ancestry. It is present in only 5% of the normal chromosomes in the Portuguese population. OBJECTIVE: To trace the origin of expanded alleles of MJD in the Indian population. METHODS: We performed CAG repeat size determination and haplotype analysis for 9 families with MJD and 263 unrelated chromosomes with unexpanded CAG sequences from the Indian population. RESULTS: All the expanded alleles were exclusively associated with the ACA haplotype in the Indian population. Interestingly, this haplotype was very common in normal alleles (40%) as compared with the Portuguese population (5%) and was significantly associated with large normal alleles (Pearson chi(2)1 = 87.1, P<.001) in the Indian population. We also observed a rare intermediate allele of MJD with the ACA haplotype but with a CAG variant instead of CAA at the sixth position in the repeat tract. CONCLUSIONS: Overrepresentation of the ACA haplotype in large normal alleles in India as compared with the Portuguese population suggests that the expansion-prone large normal alleles with the ACA haplotype may have been introduced in the Portuguese population through admixture with South Asian populations. Detailed haplotype analysis of a CAG variant within the repeat tract in an intermediate allele of MJD suggests a mechanism of gene conversion in the expansion. PMID- 15824266 TI - Dietary folate and vitamin B12 intake and cognitive decline among community dwelling older persons. AB - BACKGROUND: Deficiencies in folate and vitamin B12 have been associated with neurodegenerative disease. OBJECTIVE: To examine the association between rates of age-related cognitive change and dietary intakes of folate and vitamin B12. DESIGN: Prospective study performed from 1993 to 2002. SETTING: Geographically defined biracial community in Chicago, Ill. PARTICIPANTS: A total of 3718 residents, 65 years and older, who completed 2 to 3 cognitive assessments and a food frequency questionnaire. MAIN OUTCOME MEASURE: Change in cognitive function measured at baseline and 3-year and 6-year follow-ups, using the average z score of 4 tests: the East Boston Tests of immediate and delayed recall, the Mini Mental State Examination, and the Symbol Digit Modalities Test. RESULTS: High folate intake was associated with a faster rate of cognitive decline in mixed models adjusted for multiple risk factors. The rate of cognitive decline among persons in the top fifth of total folate intake (median, 742 microg/d) was more than twice that of those in the lowest fifth of intake (median, 186 microg/d), a statistically significant difference of 0.02 standardized unit per year (P = .002). A faster rate of cognitive decline was also associated with high folate intake from food (P for trend = .04) and with folate vitamin supplementation of more than 400 microg/d compared with nonusers (beta = -.03, P<.001). High total B12 intake was associated with slower cognitive decline only among the oldest participants. CONCLUSIONS: High intake of folate may be associated with cognitive decline in older persons. These unexpected findings call for further study of the cognitive implications of high levels of dietary folate in older populations. PMID- 15824267 TI - Acute disseminated encephalomyelitis after liver transplantation. AB - BACKGROUND: During the past 10 years, acute disseminated encephalomyelitis has been reported a few times after organ transplantation. OBJECTIVE: To report a case of acute disseminated encephalomyelitis as a complication of liver transplantation. DESIGN: Case report. SETTING: The University of North Carolina Hospital and Medical Center, Chapel Hill. Patient A 49-year-old woman admitted because of acute onset of paresthesias, sensory loss, and weakness after liver transplantation. Acute clinical presentation, results of imaging studies, and comprehensive laboratory evaluation were consistent with acute disseminated encephalomyelitis. INTERVENTIONS: High-dose intravenous corticosteroid therapy followed by maintenance oral dosing. MAIN OUTCOME MEASURES: Clinical and magnetic resonance imaging improvement. RESULTS: Corticosteroid therapy halted clinical progression, with partial resolution of lesions on magnetic resonance images of the brain and spinal cord. CONCLUSIONS: This is, to our knowledge, the first report of acute disseminated encephalomyelitis after liver transplantation. Possible pathogenic mechanisms include a cross-reactive immune response to foreign antigens present within the transplanted organ, or an inflammatory response triggered by viral infection in an immunocompromised host. PMID- 15824268 TI - Value of gradient-echo magnetic resonance imaging in the diagnosis of familial cerebral cavernous malformation. AB - BACKGROUND: Cerebral cavernous malformations (CCMs) are congenital vascular anomalies that can cause seizures, intracranial hemorrhages, focal neurological deficits, and migrainelike headaches. Magnetic resonance (MR) imaging has substantially facilitated diagnosis of CCM. It is now widely accepted that familial clustering with an autosomal dominant inheritance pattern should be suspected in cases of multiple lesions. OBJECTIVE: To determine by MR imaging the penetrance of cavernous malformations in a 3-generation family that included 5 members with typical clinical signs and diagnostic findings. METHODS: All family members underwent routine MR T1-weighted and T2-weighted spin-echo sequences in addition to MR T2-weighted gradient-echo sequences. RESULTS: Four family members had been symptomatic with either brainstem bleeding, headaches, or focal neurological signs. The gradient-echo sequences yielded a dramatically higher sensitivity with regard to lesion number and distribution. As in previous reports of familial CCM, an increase in lesion number with increasing age, changes in lesion characteristics, de novo occurrence in serial MR imaging over time, and the phenomenon of anticipation could be confirmed in this family. CONCLUSION: Magnetic resonance gradient-echo sequences should be considered the method of choice for diagnosis of familial CCM. PMID- 15824269 TI - Leigh syndrome associated with mitochondrial complex I deficiency due to a novel mutation in the NDUFS1 gene. AB - BACKGROUND: Mutations in the nuclear-encoded subunits of complex I of the mitochondrial respiratory chain are a recognized cause of Leigh syndrome (LS). Recently, 6 mutations in the NDUFS1 gene were identified in 3 families. OBJECTIVE: To describe a Spanish family with LS, complex I deficiency in muscle, and a novel mutation in the NDUFS1 gene. DESIGN: Using molecular genetic approaches, we identified the underlying molecular defect in a patient with LS with a complex I defect. PATIENT: The proband was a child who displayed the clinical features of LS. RESULTS: Muscle biochemistry results showed a complex I defect of the mitochondrial respiratory chain. Sequencing analysis of the mitochondrial DNA-encoded ND genes, the nuclear DNA-encoded NDUFV1, NDUFS1, NDUFS2, NDUFS4, NDUFS6, NDUFS7, NDUFS8, and NDUFAB1 genes, and the complex I assembly factor CIA30 gene revealed a novel homozygous L231V mutation (c.691C- >G) in the NDUFS1 gene. The parents were heterozygous carriers of the L231V mutation. CONCLUSIONS: Identifying nuclear mutations as a cause of respiratory chain disorders will enhance the possibility of prenatal diagnosis and help us understand how molecular defects can lead to complex I deficiency. PMID- 15824270 TI - L-2-hydroxyglutaric aciduria: clinical, neuroimaging, and neuropathological findings. AB - BACKGROUND: l-2-Hydroxyglutaric aciduria is a rare, infantile-onset, autosomal recessive organic aciduria affecting exclusively the central nervous system. To our knowledge, only 1 complete report of the neuropathological findings in an adult has been published. OBJECTIVE: To present the clinical, neuroimaging, and neuropathological findings of l-2-hydroxyglutaric aciduria. DESIGN: Case report. SETTING: Complexo Hospitalario de Pontevedra, Pontevedra, Spain. PATIENT: A 15 year-old boy who had early infantile-onset progressive psychomotor regression, mild choreodystonia affecting the distal part of the upper limbs, pyramidal signs, and epilepsy. RESULTS: The diagnosis of l-2-hydroxyglutaric aciduria was confirmed by the finding of highly elevated levels of l-2-hydroxyglutaric acid in the serum, urine, and cerebrospinal fluid. The neuroimaging findings showed striking confluent subcortical white matter lesions and minimal basal ganglia (pallidum, thalamic, and putaminal) abnormalities. The patient died of a spontaneous mesenteric thrombosis. The postmortem neuropathological findings showed spongiosis and cystic cavitations in subcortical white matter, with minimal abnormalities of the basal ganglia. The dentate nucleus, a structure usually affected in neuroimaging studies, showed minimal neuronal loss but was surrounded by important spongiosis and microvacuolation with astrocytic proliferation. CONCLUSIONS: This case reaffirms that l-2-hydroxyglutaric aciduria is a spongiform type of leukoencephalopathy with cystic cavitations predominating in the subcortical areas. Although the neuroimaging findings are highly characteristic of the disease, in this patient cerebellar abnormalities were minimal and dentate signal abnormalities were not present. PMID- 15824271 TI - Constipation heralding neuroborreliosis: an atypical tale of 2 patients. AB - This is a report of 2 patients with Lyme disease who initially presented with severe constipation, which progressed to ascending muscular weakness resembling acute idiopathic polyneuritis, with neuropsychiatric symptoms, severe urinary retention, and hyponatremia. These symptoms resolved following proper antibiotic therapy. PMID- 15824272 TI - Fludeoxyglucose F 18 positron emission tomography and computed tomography of a giant retroperitoneal schwannoma occurring in a patient with neurofibromatosis type 2. PMID- 15824273 TI - Babinski-Nageotte syndrome with ipsilateral hemiparesis. PMID- 15824274 TI - Primary torsion dystonia. PMID- 15824275 TI - Evaluating the evidence for multiple sclerosis as an autoimmune disease. PMID- 15824276 TI - Survival in Alzheimer disease. PMID- 15824277 TI - Brain delivery of homovanillic acid to cerebrospinal fluid during human aging. PMID- 15824278 TI - "Translational" legume biology. Models to crops. PMID- 15824279 TI - Sequencing the genespaces of Medicago truncatula and Lotus japonicus. PMID- 15824280 TI - Nematodes. Sophisticated parasites of legumes. PMID- 15824281 TI - Bridging model and crop legumes through comparative genomics. PMID- 15824282 TI - Ubiquitin-mediated proteolysis. To be in the right place at the right moment during nodule development. PMID- 15824283 TI - Peace talks and trade deals. Keys to long-term harmony in legume-microbe symbioses. PMID- 15824285 TI - Legumes as a model plant family. Genomics for food and feed report of the Cross Legume Advances Through Genomics Conference. PMID- 15824286 TI - Floral patterning in Lotus japonicus. AB - Floral patterning in Papilionoideae plants, such as pea (Pisum sativum) and Medicago truncatula, is unique in terms of floral organ number, arrangement, and initiation timing as compared to other well-studied eudicots. To investigate the molecular mechanisms involved in the floral patterning in legumes, we have analyzed two mutants, proliferating floral meristem and proliferating floral organ-2 (pfo-2), obtained by ethyl methanesulfonate mutagenesis of Lotus japonicus. These two mutants showed similar phenotypes, with indeterminate floral structures and altered floral organ identities. We have demonstrated that loss of function of LjLFY and LjUFO/Pfo is likely to be responsible for these mutant phenotypes, respectively. To dissect the regulatory network controlling the floral patterning, we cloned homologs of the ABC function genes, which control floral organ identity in Arabidopsis (Arabidopsis thaliana). We found that some of the B and C function genes were duplicated. RNA in situ hybridization showed that the C function genes were expressed transiently in the carpel, continuously in stamens, and showed complementarity with the A function genes in the heterogeneous whorl. In proliferating floral meristem and pfo-2 mutants, all B function genes were down-regulated and the expression patterns of the A and C function genes were drastically altered. We conclude that LjLFY and LjUFO/Pfo are required for the activation of B function genes and function together in the recruitment and determination of petals and stamens. Our findings suggest that gene duplication, change in expression pattern, gain or loss of functional domains, and alteration of key gene functions all contribute to the divergence of floral patterning in L. japonicus. PMID- 15824288 TI - Inhibition of plant asparagine synthetase by monoterpene cineoles. PMID- 15824289 TI - Low-molecular-weight heparins: do we have the GUSTO to identify differences between alternative formulations? PMID- 15824287 TI - A systematic proteomic study of seed filling in soybean. Establishment of high resolution two-dimensional reference maps, expression profiles, and an interactive proteome database. AB - A high-throughput proteomic approach was employed to determine the expression profile and identity of hundreds of proteins during seed filling in soybean (Glycine max) cv Maverick. Soybean seed proteins were analyzed at 2, 3, 4, 5, and 6 weeks after flowering using two-dimensional gel electrophoresis and matrix assisted laser desorption ionization time-of-flight mass spectrometry. This led to the establishment of high-resolution proteome reference maps, expression profiles of 679 spots, and corresponding matrix-assisted laser desorption ionization time-of-flight mass spectrometry spectra for each spot. Database searching with these spectra resulted in the identification of 422 proteins representing 216 nonredundant proteins. These proteins were classified into 14 major functional categories. Proteins involved in metabolism, protein destination and storage, metabolite transport, and disease/defense were the most abundant. For each functional category, a composite expression profile is presented to gain insight into legume seed physiology and the general regulation of proteins associated with each functional class. Using this approach, an overall decrease in metabolism-related proteins versus an increase in proteins associated with destination and storage was observed during seed filling. The accumulation of unknown proteins, sucrose transport and cleavage enzymes, cysteine and methionine biosynthesis enzymes, 14-3-3-like proteins, lipoxygenases, storage proteins, and allergenic proteins during seed filling is also discussed. A user-intuitive database (http://oilseedproteomics.missouri.edu) was developed to access these data for soybean and other oilseeds currently being investigated. PMID- 15824290 TI - Effect of different antilipidemic agents and diets on mortality: a systematic review. AB - BACKGROUND: Guidelines for the prevention and treatment of hyperlipidemia are often based on trials using combined clinical end points. Mortality data are the most reliable data to assess efficacy of interventions. We aimed to assess efficacy and safety of different lipid-lowering interventions based on mortality data. METHODS: We conducted a systematic search of randomized controlled trials published up to June 2003, comparing any lipid-lowering intervention with placebo or usual diet with respect to mortality. Outcome measures were mortality from all, cardiac, and noncardiovascular causes. RESULTS: A total of 97 studies met eligibility criteria, with 137,140 individuals in intervention and 138,976 individuals in control groups. Compared with control groups, risk ratios for overall mortality were 0.87 for statins (95% confidence interval [CI], 0.81 0.94), 1.00 for fibrates (95% CI, 0.91-1.11), 0.84 for resins (95% CI, 0.66 1.08), 0.96 for niacin (95% CI, 0.86-1.08), 0.77 for n-3 fatty acids (95% CI, 0.63-0.94), and 0.97 for diet (95% CI, 0.91-1.04). Compared with control groups, risk ratios for cardiac mortality indicated benefit from statins (0.78; 95% CI, 0.72-0.84), resins (0.70; 95% CI, 0.50-0.99) and n-3 fatty acids (0.68; 95% CI, 0.52-0.90). Risk ratios for noncardiovascular mortality of any intervention indicated no association when compared with control groups, with the exception of fibrates (risk ratio, 1.13; 95% CI, 1.01-1.27). CONCLUSIONS: Statins and n-3 fatty acids are the most favorable lipid-lowering interventions with reduced risks of overall and cardiac mortality. Any potential reduction in cardiac mortality from fibrates is offset by an increased risk of death from noncardiovascular causes. PMID- 15824284 TI - Sequencing and analysis of common bean ESTs. Building a foundation for functional genomics. AB - Although common bean (Phaseolus vulgaris) is the most important grain legume in the developing world for human consumption, few genomic resources exist for this species. The objectives of this research were to develop expressed sequence tag (EST) resources for common bean and assess nodule gene expression through high density macroarrays. We sequenced a total of 21,026 ESTs derived from 5 different cDNA libraries, including nitrogen-fixing root nodules, phosphorus-deficient roots, developing pods, and leaves of the Mesoamerican genotype, Negro Jamapa 81. The fifth source of ESTs was a leaf cDNA library derived from the Andean genotype, G19833. Of the total high-quality sequences, 5,703 ESTs were classified as singletons, while 10,078 were assembled into 2,226 contigs producing a nonredundant set of 7,969 different transcripts. Sequences were grouped according to 4 main categories, metabolism (34%), cell cycle and plant development (11%), interaction with the environment (19%), and unknown function (36%), and further subdivided into 15 subcategories. Comparisons to other legume EST projects suggest that an entirely different repertoire of genes is expressed in common bean nodules. Phaseolus-specific contigs, gene families, and single nucleotide polymorphisms were also identified from the EST collection. Functional aspects of individual bean organs were reflected by the 20 contigs from each library composed of the most redundant ESTs. The abundance of transcripts corresponding to selected contigs was evaluated by RNA blots to determine whether gene expression determined by laboratory methods correlated with in silico expression. Evaluation of root nodule gene expression by macroarrays and RNA blots showed that genes related to nitrogen and carbon metabolism are integrated for ureide production. Resources developed in this project provide genetic and genomic tools for an international consortium devoted to bean improvement. PMID- 15824291 TI - A randomized trial comparing 2 low-molecular-weight heparins for the outpatient treatment of deep vein thrombosis and pulmonary embolism. AB - BACKGROUND: Low-molecular-weight heparins (LMWHs) are now standard therapy for the treatment of deep vein thrombosis (DVT) and pulmonary embolism (PE). No published trials have compared LMWHs, and few studies have examined outpatient therapy for PE. Only tinzaparin sodium has demonstrated superiority to unfractionated heparin in a clinical trial. METHODS: We compared 2 LMWH products, tinzaparin and dalteparin sodium, for the treatment of acute DVT and PE in a randomized, controlled clinical trial of consecutive outpatients presenting to a venous thromboembolism service at 4 tertiary-care hospitals. Patients were treated with subcutaneous tinzaparin sodium, 175 IU/kg every 24 hours, or subcutaneous dalteparin sodium, 200 IU/kg every 24 hours, for at least 5 days. Warfarin sodium therapy was started simultaneously and continued for 90 days. The primary end point was efficacy (recurrence of venous thromboembolism); safety (bleeding) was a composite end point. RESULTS: Two hundred fifty-four patients received tinzaparin (39 with PE and 215 with DVT) and 251 received dalteparin (51 with PE and 200 with DVT). Most patients had an active malignancy or idiopathic DVT/PE. The outcome events occurred in 11 (4.4%; 95% confidence interval [CI], 2.2%-7.7%) and 15 patients (5.9%; 95% CI, 3.3%-9.5%) in the dalteparin and tinzaparin groups, respectively, including 9 and 10 recurrences, respectively, and 2 and 5 major hemorrhages, respectively (P = .44). The 95% CI on the difference of -1.5% was -5.3% to 2.4%. CONCLUSIONS: Tinzaparin and dalteparin are safe and effective for the outpatient treatment of DVT or PE. Our finding of no differences between the LMWHs based on major clinical end points means that practical issues can be the deciding factor on which drug to use. PMID- 15824292 TI - Obesity, weight change, hypertension, diuretic use, and risk of gout in men: the health professionals follow-up study. AB - BACKGROUND: Limited prospective information exists on the relation between obesity and weight change and the risk of gout. Similarly, both hypertension and diuretic use have been considered risk factors for gout; however, their independent contributions have not been established prospectively. METHODS: We prospectively examined over a 12-year period (1986-1998) the relation between adiposity, weight change, hypertension, and diuretic use and incident gout in 47,150 male participants with no history of gout at baseline. We used a supplementary questionnaire to ascertain the American College of Rheumatology criteria for gout. RESULTS: During 12 years we documented 730 confirmed incident cases of gout. Compared with men with a body mass index (BMI) of 21 to 22.9, the multivariate relative risks (RRs) of gout were 1.95 (95% confidence interval [CI], 1.44-2.65) for men with a BMI of 25 to 29.9, 2.33 (95% CI, 1.62-3.36) for men with a BMI of 30 to 34.9, and 2.97 (95% CI, 1.73-5.10) for men with a BMI of 35 or greater (P for trend <.001). Compared with men who had maintained their weight (+/-4 lb) since age 21 years, the multivariate RR of gout for men who had gained 30 lb or more since age 21 years was 1.99 (95% CI, 1.49-2.66). In contrast, the multivariate RR for men who had lost 10 lb or more since the study baseline was 0.61 (95% CI, 0.40-0.92). The multivariate RRs of gout were 2.31 (95% CI, 1.96-2.72) for the presence of hypertension and 1.77 (95% CI, 1.42-2.20) for diuretic use. CONCLUSIONS: Higher adiposity and weight gain are strong risk factors for gout in men, while weight loss is protective. Hypertension and diuretic use are also important independent risk factors for gout. PMID- 15824293 TI - Management of aortic disease in Marfan Syndrome: a decision analysis. AB - BACKGROUND: Marfan syndrome is a relatively common heritable disorder of connective tissue that affects numerous organ systems, but the most severe complication is aortic aneurysm and dissection. A variety of medical and surgical approaches are available for managing the cardiovascular complications. Our objective was to compare elective composite graft surgery, elective valve-sparing surgery, and medical management for patients with both Marfan syndrome and thoracic aortic disease on the basis of life expectancy with differing diameters of the aortic root and rate of increase in the aortic root size. METHODS: A Markov decision analysis model was constructed to compare the 2 surgical options with watchful waiting with medical therapy. RESULTS: For our base-case analysis of a 20-year-old patient with Marfan syndrome and thoracic aortic aneurysm, the aortic valve-sparing option was preferred. It extended life expectancy to 73.8 years compared with the medical treatment option (71.4 years) and with the composite graft surgery (72.7 years). Our results show that there is a better outcome for a patient with an aortic root diameter between 3.0 and 3.5 cm with early prophylactic surgery than with deferred or emergency surgery. Medical treatment was preferred when the aortic root diameter was smaller than 3.0 cm. CONCLUSIONS: Although long-term follow-up data are not yet available, it appears that advances in the technique of valve-sparing surgery have made it the preferred option to composite graft, primarily to avoid the complications of anticoagulation. Our study indicates that patients who have an aortic root diameter of larger than 3.0 cm should be considered for prophylactic aortic surgery. PMID- 15824294 TI - Effect of exercise on blood pressure in older persons: a randomized controlled trial. AB - BACKGROUND: Because of age-related differences in the cause of hypertension, it is uncertain whether current exercise guidelines for reducing blood pressure (BP) are applicable to older persons. Few exercise studies in older persons have evaluated BP changes in relation to changes in body composition or fitness. METHODS: This was a 6-month randomized controlled trial of combined aerobic and resistance training; controls followed usual care physical activity and diet advice. Participants (aged 55-75 years) had untreated systolic BP (SBP) of 130 to 159 mm Hg or diastolic BP (DBP) of 85 to 99 mm Hg. RESULTS: Fifty-one exercisers and 53 controls completed the trial. Exercisers significantly improved aerobic and strength fitness, increased lean mass, and reduced general and abdominal obesity. Mean decreases in SBP and DBP, respectively, were 5.3 and 3.7 mm Hg among exercisers and 4.5 and 1.5 mm Hg among controls (P < .001 for all). There were no significant group differences in mean SBP change from baseline (-0.8 mm Hg; P=.67). The mean DBP reduction was greater among exercisers (-2.2 mm Hg; P=.02). Aortic stiffness, indexed by aortofemoral pulse-wave velocity, was unchanged in both groups. Body composition improvements explained 8% of the SBP reduction (P = .006) and 17% of the DBP reduction (P<.001). CONCLUSIONS: A 6 month program of aerobic and resistance training lowered DBP but not SBP in older adults with mild hypertension more than in controls. The concomitant lack of improvement in aortic stiffness in exercisers suggests that older persons may be resistant to exercise-induced reductions in SBP. Body composition improvements were associated with BP reductions and may be a pathway by which exercise training improves cardiovascular health in older men and women. PMID- 15824295 TI - Changes in disability before and after myocardial infarction in older adults. AB - BACKGROUND: Disability in older adults is thought to occur primarily as a consequence of clinical disease episodes. However, the temporal relationship between clinical disease and disability has received little systematic attention. METHODS: Data from a prospective population-based study of 2812 older adults were analyzed to examine changes in disability before and after acute myocardial infarction. Disability outcomes included up to 9 yearly assessments of disability in activities in daily living, disability in basic physical functions, and disability in tasks requiring basic mobility and strength. RESULTS: A total of 279 myocardial infarctions occurred during 9 years of follow-up. After adjustment for age and sex, the average yearly increase in disability in activities of daily living and basic physical functions was not significantly greater in the 1-year period after myocardial infarction than in the 3-year period before myocardial infarction (P values >.20). Disability in basic mobility and strength showed a significantly greater increase in the year after myocardial infarction (P = .02). The results did not change after adjustment for comorbidity and chest pain or when restricted to incident cases of myocardial infarction or survivors. An additional exploratory analysis suggested that the rate of increase in some forms of disability may start to accelerate at about 1 year before the event, rather than after the event. CONCLUSIONS: The increase in disability after myocardial infarction may form a continuation of increases that occur before the event and challenge commonly held notions about the temporal relationship between clinical disease and disability. Changes in disability before acute disease episodes may be related to subclinical disease. PMID- 15824296 TI - Health insurance coverage during the years preceding medicare eligibility. AB - BACKGROUND: Adults in late middle age who lack health insurance are more likely to die or experience a decline in their overall health. Because most estimates of the uninsured are cross-sectional, the true number of individuals whose health is at risk from being uninsured is unclear. METHODS: We analyzed a nationally representative sample of 6065 US adults 51 to 57 years old who were interviewed in 1992, 1994, 1996, 1998, and 2000 as part of the Health and Retirement Study. Insurance coverage was determined at the time of each interview and classified as private, public, or uninsured. Longitudinal data were used to determine the proportion of individuals who were uninsured at any interview during the 8-year study period. RESULTS: The proportion of participants who were uninsured at the time of the 1992, 1994, 1996, 1998, and 2000 interviews was 14.3%, 10.8%, 9.7%, 8.8%, and 8.2%, respectively. People frequently transitioned between having insurance and being uninsured. As a result, despite the declining prevalence of being uninsured, the percentage who were uninsured at least once during the 8 year period rose to 23.3% by 2000; few participants (2.6%) were continuously uninsured. Only 60.1% of participants were continuously enrolled in private insurance across all 5 interviews. CONCLUSIONS: The proportion of US adults in late middle age at risk from being uninsured over a 10-year follow-up period was 2 to 3 times higher than cross-sectional estimates. At least one quarter of older adults will be uninsured at some point during the years preceding eligibility for Medicare. PMID- 15824297 TI - Obesity, regional body fat distribution, and the metabolic syndrome in older men and women. AB - BACKGROUND: The metabolic syndrome is a disorder that includes dyslipidemia, insulin resistance, and hypertension and is associated with an increased risk of diabetes and cardiovascular disease. We determined whether patterns of regional fat deposition are associated with metabolic syndrome in older adults. METHODS: A cross-sectional study was performed that included a random, population-based, volunteer sample of Medicare-eligible adults within the general communities of Pittsburgh, Pa, and Memphis, Tenn. The subjects consisted of 3035 men and women aged 70 to 79 years, of whom 41.7% were black. Metabolic syndrome was defined by Adult Treatment Panel III criteria, including serum triglyceride level, high density lipoprotein cholesterol level, glucose level, blood pressure, and waist circumference. Visceral, subcutaneous abdominal, intermuscular, and subcutaneous thigh adipose tissue was measured by computed tomography. RESULTS: Visceral adipose tissue was associated with the metabolic syndrome in men who were of normal weight (odds ratio, 95% confidence interval: 2.1, 1.6-2.9), overweight (1.8, 1.5-2.1), and obese (1.2, 1.0-1.5), and in women who were of normal weight (3.3, 2.4-4.6), overweight (2.4, 2.0-3.0), and obese (1.7, 1.4-2.1), adjusting for race. Subcutaneous abdominal adipose tissue was associated with the metabolic syndrome only in normal-weight men (1.3, 1.1-1.7). Intermuscular adipose tissue was associated with the metabolic syndrome in normal-weight (2.3, 1.6-3.5) and overweight (1.2, 1.1-1.4) men. In contrast, subcutaneous thigh adipose tissue was inversely associated with the metabolic syndrome in obese men (0.9, 0.8-1.0) and women (0.9, 0.9-1.0). CONCLUSION: In addition to general obesity, the distribution of body fat is independently associated with the metabolic syndrome in older men and women, particularly among those of normal body weight. PMID- 15824298 TI - Bleeding complications associated with combinations of aspirin, thienopyridine derivatives, and warfarin in elderly patients following acute myocardial infarction. AB - BACKGROUND: Combinations of aspirin with thienopyridine derivatives (clopidogrel bisulfate or ticlopidine hydrochloride) and/or warfarin sodium are increasingly being used in various cardiac conditions. However, little is known about the bleeding risks associated with these combinations, particularly in elderly individuals at the population level. This study estimates the bleeding risks associated with combinations of aspirin, thienopyridine derivatives, and warfarin in elderly patients. METHODS: We conducted a population-based observational cohort study using linked administrative databases. A total of 21,443 elderly survivors of acute myocardial infarction between 1996 and 2000 were studied. Patients were divided into 5 groups according to drug exposure: aspirin alone, warfarin alone, aspirin plus a thienopyridine derivative (antiplatelet combination), aspirin plus warfarin (anticoagulant combination), and aspirin plus warfarin plus a thienopyridine derivative (3-drug combination). Hospitalizations for bleeding events were examined. RESULTS: Hospitalizations for bleeding were observed in 1428 patients (7%). Compared with rates of patients receiving aspirin alone (0.03 per patient-year), rates of bleeding were higher among patients receiving the antiplatelet combination (0.07 per patient-year), the anticoagulant combination (0.08 per patient-year), and the 3-drug combination (0.09 per patient year). Compared with aspirin alone, the adjusted odds ratios (95% confidence intervals) for bleeding were 1.65 (1.02-2.73) for patients receiving the antiplatelet combination and 1.92 (1.28-2.87) for patients receiving the anticoagulant combination. Only 1 of 141 patients in the 3-drug combination group had a bleeding event. CONCLUSION: In practice, antiplatelet and anticoagulant combinations lead to modest increases in bleeding risk in elderly patients, but the overall risk is small. PMID- 15824299 TI - Concealed renal insufficiency and adverse drug reactions in elderly hospitalized patients. AB - BACKGROUND: Adverse drug reactions (ADRs) are common causes of in-hospital complications for elderly people. The purpose of the present study is to verify whether concealed renal insufficiency, that is, reduction of the estimated glomerular filtration rate (GFR) in people with normal serum creatinine levels, is a risk factor for ADRs in elderly hospitalized patients. METHODS: We used data on 11,687 hospitalized patients enrolled in the Gruppo Italiano di Farmacovigilanza nell'Anziano study. The outcomes of the study were any ADR, ADR to hydrosoluble drugs, and ADR to any other drug during the hospital stay. We compared 3 groups: normal renal function (normal serum creatinine levels and normal estimated GFRs), concealed (normal serum creatinine levels and reduced estimated GFRs), or overt (increased creatinine levels and reduced estimated GFRs) renal insufficiency. The relationship between renal function and ADR was evaluated using contingency tables and multiple regression analysis including potential confounders. RESULTS: Concealed renal insufficiency was detected in 1631 (13.9%) patients and was frequently associated with male sex and poor nutritional status. Hydrosoluble drugs were responsible for 301 of the 941 recorded ADRs. After adjusting for potential confounders, both concealed (odds ratio [OR], 1.61; 95% confidence interval [CI], 1.15-1.25) and overt (OR, 2.02; 95% CI, 1.54-2.65) renal failure were associated with ADR to hydrosoluble drugs, but not with ADR to other drugs (OR, 0.83 [95% CI, 0.65-1.08], and OR, 1.01 [95%CI, 0.83-1.23], respectively). CONCLUSION: Older hospitalized patients frequently have impaired renal function despite normal serum creatinine levels and are exposed to an increased risk of ADRs to hydrosoluble drugs. PMID- 15824300 TI - Effects of a hospitalist model on elderly patients with hip fracture. AB - BACKGROUND: Hospitalists' increased role in perioperative medicine allows for examination of their effects on surgical patients. This study examined the effects of a hospitalist service created to medically manage elderly patients with hip fracture. METHODS: During a 2-year historical cohort study of 466 patients 65 years or older admitted for surgical repair of hip fracture, we examined outcomes 1 year prior to and subsequent to the change from the standard to the hospitalist model. RESULTS: The mean (SD) time to surgery (38 [47] vs 25 [53] hours; P<.001), time from surgery to dismissal (9 [8] vs 7 [5] days; P = .04), and length of stay (10.6 [9] vs 8.4 [6] days; P<.001) were shorter in the hospitalist group. Predictors of shorter time to surgery were care by the hospitalist group (P = .002), older age (P = .01), and fall as the mechanism of fracture (P<.001), while American Society of Anesthesia scores of 3 and 4 were associated with increased time to surgery (P<.001). Receiving care by the hospitalist group (P<.001) and diagnosis of delirium (P<.001) were associated with increased chance of earlier dismissal, while admission to the intensive care unit decreased this chance (P<.001). Diagnosis of delirium was more frequent in the hospitalist group (74 [32.2%] of 230 vs 42 [17.8%] of 236; P<.001). There were no differences in inpatient deaths or 30-day readmission rates. CONCLUSION: In elderly patients with hip fracture, a hospitalist model decreased time to surgery, time from surgery to dismissal, and length of stay without adversely affecting inpatient deaths or 30-day readmission rates. PMID- 15824302 TI - Guided prescription of psychotropic medications for geriatric inpatients. AB - BACKGROUND: Inappropriate use or excessive dosing of psychotropic medications in the elderly is common and can lead to a variety of adverse drug events including falls, oversedation, and cognitive impairment. METHODS: We developed a database of psychotropic medication dosing and selection guidelines for elderly inpatients. We displayed these recommendations to physicians through a computerized order entry system at a tertiary care academic hospital. The system was activated for 2 of 4 six-week study periods in an off-on-off-on pattern. Main outcome measures were agreement with the recommended daily dose for the initial drug order, incidence of dosing at least 10-fold greater than the recommended daily dose, prescription of nonrecommended drugs, inpatient falls, altered mental status as measured by a brief nursing assessment, and hospital length of stay. RESULTS: A total of 7456 initial orders for psychotropic medications were prescribed for 3718 hospitalized elderly patients with a mean +/- SD age of 74.7 +/- 6.7 years. The intervention increased the prescription of the recommended daily dose (29% vs 19%; P<.001), reduced the incidence of 10-fold dosing (2.8% vs 5.0%; P<.001), and reduced the prescription of nonrecommended drugs (10.8% vs 7.6% of total orders; P<.001). Patients in the intervention cohort had a lower in hospital fall rate (0.28 vs 0.64 falls per 100 patient-days; P = .001). No effect on hospital length of stay or days of altered mental status was found. CONCLUSION: A geriatric decision support system for psychotropic medications increased the prescription of recommended doses, reduced the prescription of nonrecommended drugs, and was associated with fewer inpatient falls. PMID- 15824303 TI - A prescribing cascade involving cholinesterase inhibitors and anticholinergic drugs. AB - BACKGROUND: The prescribing cascade model involves the misinterpretation of an adverse reaction to 1 drug and the subsequent, potentially inappropriate prescription of a second drug. We present a new example of the prescribing cascade involving cholinesterase inhibitors and anticholinergic drugs used to manage urinary incontinence. METHODS: A population-based retrospective cohort study was carried out in Ontario, Canada. Participants included 44,884 older adults with dementia (20,491 were dispensed a cholinesterase inhibitor and 24,393 were not), enrolled between June 1, 1999, and March 31, 2002. Subjects were observed until they received an anticholinergic drug, stopped the cholinesterase inhibitor treatment, died, or the study period ended (March 31, 2003). The main outcome measure was receipt of an anticholinergic drug to manage urinary incontinence. RESULTS: After adjusting for potential confounding factors, we observed that older adults with dementia who were dispensed cholinesterase inhibitors had an increased risk of subsequently receiving an anticholinergic drug (4.5% vs 3.1%; P<.001; adjusted hazard ratio, 1.55; 95% confidence interval, 1.39-1.72), relative to those not receiving cholinesterase inhibitors. This finding was consistent in a series of subgroup analyses. CONCLUSIONS: Use of cholinesterase inhibitors is associated with an increased risk of receiving an anticholinergic drug to manage urinary incontinence. The use of an anticholinergic drug in this setting may represent a clinically important prescribing cascade. Clinicians should consider the possible contributing role of cholinesterase inhibitors in new-onset or worsening urinary incontinence and the potential risk of coprescribing cholinesterase inhibitors and anticholinergic drugs to patients with dementia. PMID- 15824304 TI - Nickel and sulfites food allergy in patients with angioedema associated with ACE Inhibitor use. PMID- 15824305 TI - Cigarette smoking, cough reflex, and respiratory tract infection. PMID- 15824306 TI - Evolutionary dynamics of olfactory receptor genes in fishes and tetrapods. AB - Olfaction, which is an important physiological function for the survival of mammals, is controlled by a large multigene family of olfactory receptor (OR) genes. Fishes also have this gene family, but the number of genes is known to be substantially smaller than in mammals. To understand the evolutionary dynamics of OR genes, we conducted a phylogenetic analysis of all functional genes identified from the genome sequences of zebrafish, pufferfish, frogs, chickens, humans, and mice. The results suggested that the most recent common ancestor between fishes and tetrapods had at least nine ancestral OR genes, and all OR genes identified were classified into nine groups, each of which originated from one ancestral gene. Eight of the nine group genes are still observed in current fish species, whereas only two group genes were found from mammalian genomes, showing that the OR gene family in fishes is much more diverse than in mammals. In mammals, however, one group of genes, gamma, expanded enormously, containing approximately 90% of the entire gene family. Interestingly, the gene groups observed in mammals or birds are nearly absent in fishes. The OR gene repertoire in frogs is as diverse as that in fishes, but the expansion of group gamma genes also occurred, indicating that the frog OR gene family has both mammal- and fish-like characters. All of these observations can be explained by the environmental change that organisms have experienced from the time of the common ancestor of all vertebrates to the present. PMID- 15824307 TI - DNA repair proteins affect the lifecycle of herpes simplex virus 1. AB - We report that herpes simplex virus 1 (HSV-1) infection can activate and exploit a cellular DNA damage response that aids viral replication in nonneuronal cells. Early in HSV-1 infection, several members of the cellular DNA damage-sensing machinery are activated and accumulate at sites of viral DNA replication. When this cellular response is abrogated, formation of HSV-1 replication centers is retarded, and viral production is compromised. In neurons, HSV-1 replication centers fail to mature, and the DNA damage response is not initiated. These data suggest that the failure of neurons to mount a DNA damage response to HSV-1 may contribute to the establishment of latency. PMID- 15824308 TI - Delayed maturation of receptive field center/surround mechanisms in V2. AB - Neurons in the adult visual cortex are capable of integrating signals over a large area that surrounds their classic receptive field (RF), and this ability of cortical neurons is thought to be intimately involved in perceptual binding. It is not known, however, at what age these long-range signal interactions emerge. Here, we report that qualitatively adult-like center/surround interactions are already present in the primary visual cortex as early as postnatal day 14 in macaque monkeys. However, the RF surrounds of visual area 2 (V2) neurons were largely absent until 4 weeks of age and, as late as 8 weeks of age, center/surround signal interactions in V2 neurons were immature. Our results suggest that the cortical circuits underlying the RF center/surround of individual neurons mature considerably later in V2 than in the primary visual cortex and give critical evidence for the hypothesis that the functional maturation of the primate visual brain proceeds in a hierarchical manner. PMID- 15824310 TI - Herpes simplex virus 1 infected cell protein 0 forms a complex with CIN85 and Cbl and mediates the degradation of EGF receptor from cell surfaces. AB - Infected cell protein 0 (ICP0) is a 775-residue multifunctional herpes simplex virus protein associated with numerous functions related to transactivation of gene expression and repression of host defenses to infection. We report that an uncharted domain of ICP0 located between residues 245 and 510 contains multiple SH3 domain binding motifs similar to those required for binding to CIN85, the M(r) 85,000 protein that interacts with Cbl. CIN85 and Cbl are involved in endocytosis and negative regulation of numerous receptor tyrosine kinases. We report that ICP0 binds CIN85 in a reciprocal manner and that the complexes pulled down by ICP0 also contain Cbl. We tested the role of ICP0 in the down-regulation of receptor tyrosine kinases by using epidermal growth factor receptor (EGFR) as a prototypic receptor. In transfection assays, ICP0, in the absence of other viral genes, down-regulated EGF-dependent expression of a reporter gene (luciferase). ICP0 also down-regulated both total and cell surface levels of EGFR in EGF-independent manner. In wild-type virus-infected cells, the surface levels of EGFR were also decreased in the absence of EGF stimulation. Stimulation by EGF enhanced the decrease in surface EGFR. We conclude that ICP0 encodes SH3 domain binding sites that function to down-regulate signaling pathways associated with receptor tyrosine kinases. The results suggest that ICP0 precludes signaling to the infected cells through the receptor tyrosine kinases. PMID- 15824309 TI - Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence. AB - Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-). PMID- 15824311 TI - Observing spontaneous branch migration of Holliday junctions one step at a time. AB - Genetic recombination occurs between homologous DNA molecules via a four-way (Holliday) junction intermediate. This ancient and ubiquitous process is important for the repair of double-stranded breaks, the restart of stalled replication forks, and the creation of genetic diversity. Once formed, the four way junction alone can undergo the stepwise exchange of base pairs known as spontaneous branch migration. Conventional ensemble assays, useful for finding average migration rates over long sequences, have been unable to examine the affect of sequence and structure on the migration process. Here, we present a single-molecule spontaneous branch migration assay with single-base pair resolution in a study of individual DNA junctions that can undergo one step of migration. Junctions exhibit markedly different dynamics of exchange between stacking conformers depending on the point of strand exchange, allowing the moment at which branch migration occurs to be detected. The free energy landscape of spontaneous branch migration is found to be highly nonuniform and governed by two types of sequence-dependent barriers, with unmediated local migration being up to 10 times more rapid than the previously deduced average rate. PMID- 15824312 TI - Evolution and development of facial bone morphology in threespine sticklebacks. AB - How do developmental mechanisms evolve to control changing skeletal morphology, the shapes and sizes of individual bones? We address this question with studies of the opercle (OP), a large facial bone that has undergone marked morphological evolution in the ray-finned fish. Attributes for developmental analysis motivated us to examine how OP shape and size evolve and develop in threespine sticklebacks, a model system for understanding vertebrate evolution. We find that when Alaskan anadromous fish take up permanent residence in lakes, they evolve smaller and reshaped OPs. The change is a reduction in the amount of bone laid down along one body axis, and it arises at or shortly after the onset of OP development. A quantitative trait locus is present on linkage group 19 that contributes in a major way to this phenotype. PMID- 15824314 TI - Local and long-range stability in tandemly arrayed tetratricopeptide repeats. AB - The tetratricopeptide repeat (TPR) is a 34-aa alpha-helical motif that occurs in tandem arrays in a variety of different proteins. In natural proteins, the number of TPR motifs ranges from 3 to 16 or more. These arrays function as molecular scaffolds and frequently mediate protein-protein interactions. We have shown that correctly folded TPR domain proteins, exhibiting the typical helix-turn-helix fold, can be designed by arraying tandem repeats of an idealized TPR consensus motif. To date, three designed proteins, CTPR1, CTPR2, and CTPR3 (consensus TPR number of repeats) have been characterized. Their high-resolution crystal structures show that the designed proteins indeed adopt the typical TPR fold, which is specified by the correct positioning of key residues. Here, we present a study of the thermodynamic properties and folding kinetics of this set of designed proteins. Chemical denaturation, monitored by CD and fluorescence, was used to assess the folding and global stability of each protein. NMR-detected amide proton exchange was used to investigate the stability of each construct at a residue-specific level. The results of these studies reveal a stable core, which defines the intrinsic stability of an individual TPR motif. The results also show the relationship between the number of tandem repeats and the overall stability and folding of the protein. PMID- 15824313 TI - Hemoglobin conformation couples erythrocyte S-nitrosothiol content to O2 gradients. AB - It is proposed that the bond between nitric oxide (NO) and the Hb thiol Cys beta(93) (SNOHb) is favored when hemoglobin (Hb) is in the relaxed (R, oxygenated) conformation, and that deoxygenation to tense (T) state destabilizes the SNOHb bond, allowing transfer of NO from Hb to form other (vasoactive) S nitrosothiols (SNOs). However, it has not previously been possible to measure SNOHb without extensive Hb preparation, altering its allostery and SNO distribution. Here, we have validated an assay for SNOHb that uses carbon monoxide (CO) and cuprous chloride (CuCl)-saturated Cys. This assay is specific for SNOs and sensitive to 2-5 pmol. Uniquely, it measures the total SNO content of unmodified erythrocytes (RBCs) (SNO(RBC)), preserving Hb allostery. In room air, the ratio of SNO(RBC) to Hb in intact RBCs is stable over time, but there is a logarithmic loss of SNO(RBC) with oxyHb desaturation (slope, 0.043). This decay is accelerated by extraerythrocytic thiol (slope, 0.089; P < 0.001). SNO(RBC) stability is uncoupled from O(2) tension when Hb is locked in the R state by CO pretreatment. Also, SNO(RBC) is increased approximately 20-fold in human septic shock (P = 0.002) and the O(2)-dependent vasoactivity of RBCs is affected profoundly by SNO content in a murine lung bioassay. These data demonstrate that SNO content and O(2) saturation are tightly coupled in intact RBCs and that this coupling is likely to be of pathophysiological significance. PMID- 15824315 TI - Uncoupling of the functions of the Arabidopsis VIP1 protein in transient and stable plant genetic transformation by Agrobacterium. AB - Agrobacterium-mediated genetic transformation of plants, a unique example of transkingdom DNA transfer, requires the presence of several proteins encoded by the host cell. One such cellular factor is VIP1, an Arabidopsis protein proposed to interact with and facilitate import of the bacterial DNA-protein transport (T) complexes into the plant cell nucleus. Thus, VIP1 is required for transient expression of the bacterial DNA, an early step in the transformation process. However, the role of VIP1 in subsequent transformation events leading to the stable expression of bacterial DNA was unexplored. Here, we used reverse genetics to dissect VIP1 functionally and demonstrate its involvement in the stable genetic transformation of Arabidopsis plants by Agrobacterium. Our data indicate that the ability of VIP1 to interact with the VirE2 protein component of the T complex and localize to the cell nucleus is sufficient for transient genetic transformation, whereas its ability to form homomultimers and interact with the host cell H2A histone in planta is required for tumorigenesis and, by implication, stable genetic transformation. PMID- 15824316 TI - Heterotopic expression of MPF2 is the key to the evolution of the Chinese lantern of Physalis, a morphological novelty in Solanaceae. AB - Morphological novelties arise through changes in development, but the underlying causes of such changes are largely unknown. In the genus Physalis, sepals resume growth after pollination to encapsulate the mature fruit, forming the "Chinese lantern," a trait also termed inflated-calyx syndrome (ICS). STMADS16, which encodes a MADS-box transcription factor, is expressed only in vegetative tissues in Solanum tuberosum. Its ortholog in Physalis pubescens, MPF2, is expressed in floral tissues. Knockdown of MPF2 function in Physalis by RNA interference (RNAi) reveals that MPF2 function is essential for the development of the ICS. The phenotypes of transgenic S. tuberosum plants that overexpress MPF2 or STMADS16 corroborate these findings: these plants display enlarged sepals. Although heterotopic expression of MPF2 is crucial for ICS, remarkably, fertilization is also required. Although the ICS is less prominent or absent in the knockdown transgenic plants, epidermal cells are larger, suggesting that MPF2 exerts its function by inhibiting cell elongation and promoting cell division. In addition, severely affected Physalis knockdown lines are male sterile. Thus, heterotopic expression of MPF2 in floral tissues is involved in two novel traits: expression of the ICS and control of male fertility. Sequence differences between the promoter regions of the MPF2 and STMADS16 genes perhaps reflect exposure to different selection pressures during evolution, and correlate with the observed differences in their expression patterns. In any case, the effects of heterotopic expression of MPF2 underline the importance of recruitment of preexisting transcription factors in the evolution of novel floral traits. PMID- 15824317 TI - Xantha-l encodes a membrane subunit of the aerobic Mg-protoporphyrin IX monomethyl ester cyclase involved in chlorophyll biosynthesis. AB - Chlorophyll biosynthesis is a process involving approximately 20 different enzymatic steps. Half of these steps are common to the biosynthesis of other tetrapyrroles, such as heme. One of the least understood enzymatic steps is formation of the isocyclic ring, which is a characteristic feature of all (bacterio)chlorophyll molecules. In chloroplasts, formation of the isocyclic ring is an aerobic reaction catalyzed by Mg-protoporphyrin IX monomethyl ester cyclase. An in vitro assay for the aerobic cyclase reaction required membrane bound and soluble components from the chloroplasts. Extracts from barley (Hordeum vulgare L.) mutants at the Xantha-l and Viridis-k loci showed no cyclase activity. Fractionation of isolated plastids by Percoll gradient centrifugation showed that xantha-l and viridis-k mutants are defective in components associated with chloroplast membranes. The Xantha-l gene, corresponding to Arabidopsis thaliana CHL27, Rubrivivax gelatinosus acsF, Chlamydomonas reinhardtii CRD1, and CTH1 and situated at the short arm of barley chromosome 3 (3H), was cloned, and the mutations in xantha-l(35), xantha-l(81), and xantha-l(82) were characterized. This finding connected biochemical and genetic data because it demonstrated that Xantha-l encodes a membrane-bound cyclase subunit. The evidence suggests that the aerobic cyclase requires at least one soluble and two membrane-bound components. PMID- 15824318 TI - Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability. AB - Several mutations in PTEN-induced putative kinase 1 (PINK1) gene have been reported to be associated with recessive parkinsonism. The encoded protein is predicted to be a Ser/Thr protein kinase targeted to mitochondria. In this study, we have investigated the effects of mutations on PINK1 kinase activity in vitro and on expression levels and localization in mammalian cells. We chose to examine two point mutations: G309D, which was originally reported to be stable and properly localized in cells and L347P, which is of interest because it is present at an appreciable carrier frequency in the Philippines. We were able to confirm kinase activity and produce artificial "kinase-dead" mutants that are stable but lack activity. The L347P mutation grossly destabilizes PINK1 and drastically reduces kinase activity, whereas G309D has much more modest effects on these parameters in vitro. This finding is in line with predictions based on homology modeling. We also examined the localization of PINK1 in transfected mammalian cells by using constructs that were tagged with myc or GFP at either end of the protein. These results show that PINK1 is processed at the N terminus in a manner consistent with mitochondrial import, but the mature protein also exists in the cytosol. The physiological relevance of this observation is not yet clear, but it implies that a portion of PINK1 may be exported after processing in the mitochondria. PMID- 15824319 TI - Absence of the DNA-/RNA-binding protein MSY2 results in male and female infertility. AB - MSY2, a germ-cell-specific member of the Y-box family of DNA-/RNA-binding proteins, is proposed to function as a coactivator of transcription in the nucleus and to stabilize and store maternal and paternal mRNAs in the cytoplasm. In mice lacking Msy2, a normal Mendelian ratio is observed after matings between heterozygotes with equal numbers of phenotypically normal but sterile male and female homozygotes (Msy2-/-). Spermatogenesis is disrupted in postmeiotic null germ cells with many misshapen and multinucleated spermatids, and no spermatozoa are detected in the epididymis. Apoptosis is increased in the testes of homozygotes, and real-time RT-PCR assays reveal large reductions in the mRNA levels of postmeiotic male germ cell mRNAs and smaller reductions of meiotic germ cell transcripts. In females, there is no apparent decrease in either the number of follicles or their morphology in ovaries obtained from 2- and 8-day-old Msy2-/ mice. In contrast, follicle number and progression are reduced in 21-day-old Msy2-/- ovaries. In adult Msy2-/- females, oocyte loss increases, anovulation is observed, and multiple oocyte and follicle defects are seen. Thus, Msy2 represents one of a small number of germ-cell-specific genes whose deletion leads to the disruption of both spermatogenesis and oogenesis. PMID- 15824320 TI - An Early Pleistocene hominin mandible from Atapuerca-TD6, Spain. AB - We present a mandible recovered in 2003 from the Aurora Stratum of the TD6 level of the Gran Dolina site (Sierra de Atapuerca, northern Spain). The specimen, catalogued as ATD6-96, adds to the hominin sample recovered from this site in 1994-1996, and assigned to Homo antecessor. ATD6-96 is the left half of a gracile mandible belonging to a probably female adult individual with premolars and molars in place. This mandible shows a primitive structural pattern shared with all African and Asian Homo species. However, it is small and exhibits a remarkable gracility, a trait shared only with the Early and Middle Pleistocene Chinese hominins. Furthermore, none of the mandibular features considered apomorphic in the European Middle and Early Upper Pleistocene hominin lineage are present in ATD6-96. This evidence reinforces the taxonomic identity of H. antecessor and is consistent with the hypothesis of a close relationship between this species and Homo sapiens. PMID- 15824321 TI - Nonmammalian gonadotropin-releasing hormone molecules in the brain of promoter transgenic rats. AB - Mammalian gonadotropin-releasing hormone (GnRH1) and nonmammalian immunoreactive GnRH subtypes were examined in transgenic rats carrying an enhanced GFP (EGFP) reporter gene driven by a rat GnRH1 promoter. Double-label immunocytochemistry was performed on EGFP(+)/GnRH1 brain sections by using antisera against GnRH1, GnRH2 (chicken II), GnRH3 (salmon), or seabream GnRH. EGFP(+)/GnRH1 neurons were in the septal-preoptic hypothalamus but not in the midbrain, consistent with GnRH1-immunopositive neurons in WT rats. Apparent coexpression of EGFP(+)/GnRH1 with other GnRH subtypes was observed. All EGFP(+) neurons in the septal-preoptic hypothalamus were GnRH1-immunopositive. However, only approximately 80% of GnRH1 immunopositive neurons were EGFP(+), which awaits further elucidation. GnRH subtypes-immunopositive fibers and EGFP(+)/GnRH1 fibers were conspicuous in the organum vasculosum of the lamina terminalis, median eminence, and surrounding the ependymal walls of the third ventricle and the aqueduct in the midbrain. These results demonstrate that the expression of the EGFP-GnRH1 transgene is restricted to the bona fide GnRH1 population and provide clear morphological evidence supporting the existence of GnRH1 neuronal subpopulations in the septal-preoptic hypothalamus, which might be driven by different segments of the GnRH promoter. This genetic construct permits analyses of promoter usage in GnRH neurons, and our histochemical approaches open questions about functional relations among isoforms of this peptide, which regulates reproductive physiology in its behavioral and endocrine aspects. PMID- 15824322 TI - Full deacylation of polyethylenimine dramatically boosts its gene delivery efficiency and specificity to mouse lung. AB - High-molecular-mass polyethylenimines (PEIs) are widely used vectors for nucleic acid delivery. We found that removal of the residual N-acyl moieties from commercial linear 25-kDa PEI enhanced its plasmid DNA delivery efficiency 21 times in vitro, as well as 10,000 times in mice with a concomitant 1,500-fold enhancement in lung specificity. Several additional linear PEIs were synthesized by acid-catalyzed hydrolysis of poly(2-ethyl-2-oxazoline), yielding the pure polycations. PEI87 and PEI217 exhibited the highest efficiency in vitro: 115-fold and 6-fold above those of the commercial and deacylated PEI25s, respectively; moreover, PEI87 delivered DNA to mouse lung as efficiently as the pure PEI25 but at a lower concentration and with a 200-fold lung specificity. These improvements stem from an increase in the number of protonatable nitrogens, which presumably results in a tighter condensation of plasmid DNA and a better endosomal escape of the PEI/DNA complexes. As a validation of the potential of such linear, fully deacylated PEIs in gene therapy for lung diseases, systemic delivery in mice of the complexes of a short interfering RNA (siRNA) against a model gene, firefly luciferase, and PEI25 or PEI87 afforded a 77% and 93% suppression of the gene expression in the lungs, respectively. Furthermore, a polyplex of a siRNA against the influenza viral nucleocapsid protein gene and PEI87 resulted in a 94% drop of virus titers in the lungs of influenza-infected animals. PMID- 15824323 TI - Efficient cross-priming of tumor antigen-specific T cells by dendritic cells sensitized with diverse anti-MICA opsonized tumor cells. AB - Dendritic cells (DCs) have the capacity to prime tumor-specific T cell responses and are considered as potentially effective vaccines for immunotherapy of cancer. Critical parameters in the development of DC vaccines are the source of tumor antigen (TA) and the mode of DC-loading. Whole tumor cells contain complex assortments of TA, which has been exploited to enhance cross-presentation to CD8 T cells by DCs loaded with anti-syndecan mAb-opsonized myeloma cells. This approach may be broadly improved by targeting the MHC class I chain-related protein A (MICA), which is frequently and abundantly expressed on most if not all types of epithelial cancers but not in normal tissues except intestinal mucosa. Loading of DC with anti-MICA mAb-coated breast, melanoma, or ovarian tumor lines or uncultured ovarian cancer cells efficiently promoted TA cross-presentation and priming of multivalent anti-tumor CD8 and CD4 T cell responses. These were of substantially greater breadth and magnitude than those of T cells primed by peptide-pulsed or apoptotic tumor cell-loaded DCs. These results may advance DC vaccine development and provide a platform for adoptive T cell therapy and TA discovery. These results further suggest that antibody targeting of MICA might be applicable to elicit T cell immunity against tumors of diverse tissue origins in cancer patients. PMID- 15824324 TI - Evolutionary emergence of size-structured food webs. AB - Explaining the structure of terrestrial and aquatic food webs remains one of the most important challenges of ecological theory. Most existing models use emergent properties of food webs, such as diversity and connectance as parameters, to determine other food-web descriptors. Lower-level processes, in particular adaptation (whether by behavioral, developmental, or evolutionary mechanisms), are usually not considered. Here, we show that complex, realistic food webs may emerge by evolution from a single ancestor based on very simple ecological and evolutionary rules. In our model, adaptation acts on body size, whose impact on the metabolism and interactions of organisms is well established. Based on parameters defined at the organism scale, the model predicts emergent properties at the food-web scale. Variations of two key parameters (width of consumption niche and competition intensity) allow very different food-web structures and functionings to emerge, which are similar to those observed in some of the best documented food webs. PMID- 15824326 TI - The failed lumbar puncture. PMID- 15824325 TI - Two essential but distinct functions of the mammalian abasic endonuclease. AB - The mammalian abasic endonuclease, APE1, has two distinct roles in the repair of oxidative DNA damage and in gene regulation. Here we show that both functions are essential for cell survival. Deletion of the APE1 gene causes embryonic lethality in mice, and no nullizygous embryo fibroblasts have been isolated. We have now established nullizygous embryo fibroblast lines from APE1(-/-) mouse embryos that are transgenic with the "floxed" human APE1 (hAPE1) gene. Removal of hAPE1 by Cre expression through nuclear microinjection elicited apoptosis in these cells within 24 h, which was blocked by coinjection of the wild-type hAPE1 gene. In contrast, mutant hAPE1 alleles, lacking either the DNA repair or acetylation mediated gene regulatory function, could not prevent apoptosis, although the combination of these two mutants complemented APE deficiency induced by Cre. These results indicate that distinct and separable functions of APE1 are both essential for mammalian cells even in vitro and provide the evidence that mammalian cells, unlike yeast or Escherichia coli, absolutely require APE for survival, presumably to protect against spontaneous oxidative DNA damage. PMID- 15824327 TI - Cognitive benefits of sleep and their loss due to sleep deprivation. PMID- 15824328 TI - A multilayered approach to studying cortical malformations: EEG-fMRI. PMID- 15824329 TI - Treating fatigue in patients with MS: one step forward, one step back. PMID- 15824330 TI - Spinocerebellar ataxia type 14: opening a new door in dominant ataxia research? PMID- 15824332 TI - Conventional wisdom vs reality in stroke prevention. PMID- 15824331 TI - Adding aspirin to clopidogrel after TIA and ischemic stroke: benefits do not match risks. AB - Antiplatelet therapy is effective for reducing the risk of recurrent stroke and other serious vascular events in patients with recent TIA and ischemic stroke. Effective antiplatelet agents include aspirin, ticlopidine, clopidogrel, dipyridamole, and the combination of aspirin and dipyridamole. The combination of aspirin and clopidogrel is more effective than aspirin in patients with acute coronary syndrome but is more hazardous than clopidogrel alone in patients with recent TIA and ischemic stroke. Further trials are needed to determine whether the combination of aspirin and clopidogrel may have a role immediately after TIA and ischemic stroke in patients with symptomatic large artery atherothromboembolism and continued for approximately 3 months before switching to less hazardous antiplatelet regimens. PMID- 15824333 TI - Cortical/subcortical BOLD changes associated with epileptic discharges: an EEG fMRI study at 3 T. AB - BACKGROUND: Malformations of cortical development have characteristic interictal discharges, yet the mechanisms of generation of these discharges are not known in humans. Interictal discharges in malformations of cortical development were studied with EEG-fMRI. METHODS: Six subjects with malformations of cortical development and seizures were studied using spike-triggered fMRI at 3 T. The blood oxygen level-dependent (BOLD) signal changes associated with interictal discharges were measured. RESULTS: All subjects showed spike-related BOLD signal changes. In four subjects, the signal increases were seen in the lesion, and in four subjects, decreases were seen surrounding the lesion. Five subjects had BOLD signal changes at distant cortical sites and three had subcortical changes (basal ganglia, reticular formation, or thalamic). CONCLUSION: BOLD signal changes may be directly correlated with overall synaptic activity. Changes were found in and around the lesion of malformations of cortical development and in distant cortical and subcortical structures. The results suggest that EEG-fMRI studies might help elucidate the mechanisms of epileptic discharges in humans. PMID- 15824334 TI - Case-control study of SUDEP. AB - OBJECTIVE: To examine the influence of various factors on the risk of sudden unexpected death in epilepsy (SUDEP). METHODS: The authors investigated 154 cases in which a postmortem examination was performed. Each case had four controls with epilepsy from the community, matched for age and geographic location. Backward stepwise conditional logistic regression analysis was performed and odds ratios for risk and protection were determined. RESULTS: The risk of SUDEP was increased with a history of generalized tonic-clonic seizures in the previous 3 months (odds ratio [OR]: 13.8, 95% CI: 6.6 to 29.1). The presence of supervision at night was found to be protective (OR: 0.4, 95% CI: 0.2 to 0.8) when a supervising individual shared the same bedroom or when special precautions such as a listening device were employed (OR: 0.1, 95% CI: 0.0 to 0.3). CONCLUSION: This work lends support to the view that SUDEP is a seizure-related phenomenon and that control of tonic-clonic seizures is important in its prevention. Nocturnal supervision seems to protect against SUDEP. PMID- 15824335 TI - Risk of Stevens-Johnson syndrome and toxic epidermal necrolysis in new users of antiepileptics. AB - BACKGROUND: Estimates of risk of Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) associated with some antiepileptic drugs (AEDs) have used denominators based on the number of prescriptions or daily doses. Because the risk of SJS is highest in new users of drugs, the use of denominators reflective of all users can lead to low estimates of risk associated with drugs. In this study, risk in new users is assessed. METHODS: Data on all hospitalized patients with SJS and TEN with use of carbamazepine (CBZ), lamotrigine (LTG), phenobarbital (PHB), phenytoin (PHT), or valproic acid (VPA) were obtained from the German Registry for Serious Cutaneous Reactions. For 1998-2001, the numbers of new users were estimated from number of dispensed prescriptions in Germany, the average prescribed doses, and the duration of use in the Mediplus database (IMS Health) Germany, and assumptions that relate new use to growth in national dispensings. To minimize the probability of underestimating risk in new users, conservative estimates of new use that were somewhat lower than predicted from national prescription data were used. RESULTS: More than 90% of SJS and TEN cases occurred in the first 63 days of AED use. Over the 4 years, increases in dispensing were 5% for CBZ, 65% for LTG, 6% for PHB, -16% for PHT, and 26% for VPA. Across a range of assumptions about frequency of incident use, the risk estimates vary between 1 and 10 per 10,000 new users for CBZ, LTG, PHT, and PHY and were consistently lower for VPA. CONCLUSION: Across a range of assumptions used, the risk of hospitalization for Stevens-Johnson syndrome or toxic epidermal necrolysis in new users is low for carbamazepine, lamotrigine, phenytoin, phenobarbital, and valproic acid. Because conservative incidence use fractions were used, it is likely that some risks were overestimated. PMID- 15824336 TI - MRI of segmental zoster paresis. PMID- 15824337 TI - Modafinil for fatigue in MS: a randomized placebo-controlled double-blind study. AB - OBJECTIVE: To assess whether modafinil, a wakefulness-promoting agent, is useful for fatigue in patients with multiple sclerosis (MS). METHODS: Patients with MS with stable disability, and a baseline score of 45 or more on the Modified Fatigue Impact Scale (MFIS), were eligible for the 5-week randomized, double blind, placebo-controlled, parallel group study. The initial daily dose of modafinil was 200 mg for 1 week. Depending on tolerance, the dose was increased by 100 mg every week up to 400 mg/day and remained unchanged between day 21 and day 35. The primary outcome variable was the change of MFIS score at day 35. RESULTS: A total of 115 patients with MS were enrolled in the study and in the intention to treat analysis. The mean MFIS score at baseline was 63 +/- 9 in the placebo group and 63 +/- 10 in the modafinil group. MFIS scores improved between day 0 and day 35 in both placebo-treated and modafinil-treated groups, but no significant difference was detected between the two groups. There was no major safety concern. CONCLUSIONS: There was no improvement of fatigue in patients with multiple sclerosis treated with modafinil vs placebo according to the Modified Fatigue Impact Scale. PMID- 15824338 TI - Multiple Sclerosis Severity Score: using disability and disease duration to rate disease severity. AB - BACKGROUND: There is no consensus method for determining progression of disability in patients with multiple sclerosis (MS) when each patient has had only a single assessment in the course of the disease. METHODS: Using data from two large longitudinal databases, the authors tested whether cross-sectional disability assessments are representative of disease severity as a whole. An algorithm, the Multiple Sclerosis Severity Score (MSSS), which relates scores on the Expanded Disability Status Scale (EDSS) to the distribution of disability in patients with comparable disease durations, was devised and then applied to a collection of 9,892 patients from 11 countries to create the Global MSSS. In order to compare different methods of detecting such effects the authors simulated the effects of a genetic factor on disability. RESULTS: Cross-sectional EDSS measurements made after the first year were representative of overall disease severity. The MSSS was more powerful than the other methods the authors tested for detecting different rates of disease progression. CONCLUSION: The Multiple Sclerosis Severity Score (MSSS) is a powerful method for comparing disease progression using single assessment data. The Global MSSS can be used as a reference table for future disability comparisons. While useful for comparing groups of patients, disease fluctuation precludes its use as a predictor of future disability in an individual. PMID- 15824339 TI - Evidence of increased oxidative damage in subjects with mild cognitive impairment. AB - OBJECTIVE: To determine if increased levels of oxidative damage are present in the brains of persons with mild cognitive impairment (MCI), a condition that often precedes Alzheimer disease (AD). METHODS: The authors assessed the amount of protein carbonyls, thiobarbituric acid-reactive substances (TBARS), and malondialdehyde in the superior and middle temporal gyri (SMTG) and cerebellum of short postmortem interval and longitudinally evaluated normal subjects and those with MCI and early AD. RESULTS: Elevated levels of protein carbonyls (approximately 25%), malondialdehyde (approximately 60%), and TBARS (approximately 210%) were observed in the SMTG of individuals with MCI and early AD vs normal control subjects. The elevation in TBARS was associated with the numbers of neuritic but not diffuse plaques. Levels of protein carbonyls increased as delayed verbal memory performance declined. CONCLUSION: Oxidative damage occurs in the brain of subjects with mild cognitive impairment, suggesting that oxidative damage may be one of the earliest events in the onset and progression of Alzheimer disease. PMID- 15824340 TI - Association between mild parkinsonian signs and mild cognitive impairment in a community. AB - BACKGROUND: Mild parkinsonian signs (MPS) are associated with prevalent and incident dementia but it is not known whether they are associated with mild cognitive impairment (MCI). OBJECTIVE: To determine whether MPS and specific MPS (changes in axial function, rigidity, tremor) are associated with MCI in nondemented community-dwelling older people in northern Manhattan, NY. METHODS: Participants underwent neurologic assessment, including a modified motor portion of the Unified Parkinson Disease Rating Scale. MCI was diagnosed in nondemented participants who had cognitive impairment based on neuropsychological testing and no functional impairment. Participants with MCI were classified as having MCI with memory impairment (MCI+M) vs MCI without memory impairment (MCI-M). RESULTS: MCI was present in 608 (27.3%) of 2,230 participants, including 255 participants with MCI+M and 353 with MCI-M; 1,622 participants did not have MCI. MPS were present in 369 (16.5%) of 2,230 participants. In a univariate logistic regression model, odds of MCI+M (vs no MCI) were 51% higher in participants with MPS compared to those with no MPS (OR = 1.51, 95% CI = 1.09 to 2.09, p = 0.01). Multivariate models yielded similar results (OR = 1.45, 95% CI = 1.03 to 2.05, p = 0.03). Rigidity was present in a higher proportion of participants with MCI+M compared to participants without MCI. CONCLUSIONS: Mild parkinsonian signs, especially rigidity, are associated with amnestic mild cognitive impairment. Mild parkinsonian signs and mild cognitive impairment may share similar pathogeneses. Whether this involves Alzheimer-type pathology, Lewy bodies, or vascular changes in the basal ganglia or basal ganglia circuitry deserves further investigation in postmortem studies. PMID- 15824341 TI - Acute akinesia in Parkinson disease. AB - OBJECTIVE: To assess acute akinesia in patients with Parkinson disease (PD) ("acute akinesia" defined as a sudden deterioration in motor performance that persists for > or =48 hours despite treatment). METHODS: The study population was a cohort of 675 patients followed regularly for 12 years in the authors' outpatient clinic. All patients were studied when acute akinesia led to hospitalization. Unified Parkinson's Disease Rating Scale (UPDRS) scores were rated during the akinetic state and compared with ratings obtained 1.6 +/- 0.9 months before the onset or after recovery. RESULTS: Twenty-six patients developed acute akinesia; in 17 of the 26 patients, new akinetic symptoms first manifested at the onset of an infectious disease or after surgery and appeared unrelated to changes in treatment or altered levodopa kinetics. In nine patients, acute akinesia developed concurrently with gastrointestinal diseases or drug manipulations showed features of neuroleptic malignant syndrome. Acute akinesia severe enough to increase the UPDRS Motor Subscale score by 31.4 +/- 12.8 appeared within 2 to 3 days and persisted for 11.2 +/- 6.2 days despite attempts to increase the dopaminergic drug dose or administer continuous subcutaneous apomorphine infusion. Symptomatic recovery began 4 to 26 days after the onset of acute akinesia and appeared incomplete in 10 patients. Four patients of 26 died despite treatment. Levodopa kinetics were normal in all patients without gastrointestinal disease and in one patient with gastric stasis. CONCLUSIONS: Acute akinesia is a life-threatening complication of Parkinson disease (PD). It is unlike the "wearing-off" phenomenon that occurs when dopaminergic drug levels decline and responds to dopaminergic rescue drugs. Acute akinesia may be a clinical entity distinct from the previously described PD motor fluctuations. PMID- 15824342 TI - High-field, T2 reversed MRI of the hippocampus in transient global amnesia. AB - OBJECTIVE: To investigate the fine structural integrity of the hippocampus in patients with transient global amnesia (TGA) using high-resolution T2 reversed (T2R) MRI. METHODS: The authors performed detailed structural analysis of the hippocampus in 15 patients who had recovered from an episode of TGA and 150 randomly recruited normal volunteers across the adult age spectrum using high resolution T2R MRI obtained on a 3.0-T system. An additional 100 subjects, with stroke or tumor, were similarly studied and served as disease controls. RESULTS: The overall incidence of hippocampal cavities detected in normal volunteers increased with age but never exceeded 40%, whereas the incidence in disease control group was 31%. They were always unilateral. In contrast, cavities were found in all 15 patients with TGA (100%), an incidence higher than in normal or disease controls (p < 0.05; Ryan's multiple comparison test), and were bilateral in eight patients (53%). The cavities in all but one of the normal volunteers (99%) and all disease controls (100%) were crescent shaped and < or =2 mm in width. The cavities in 14 of 15 patients with TGA (93%) were considerably larger (>3 mm in width), and five of the patients had giant cavities (>5 mm in width). Most of the cavities in patients with TGA had a rounded shape and resembled pathologic cavities described in specimens of hypoxia-related CA1 necrosis. CONCLUSION: The data indicate that transient global amnesia may not be a benign entity. Delayed neuronal loss within CA1 area of Lorente de No may represent its important sequel. PMID- 15824343 TI - The unpleasantness of tonic pain is encoded by the insular cortex. AB - OBJECTIVE: Muscle pain differs from skin pain with respect to quality, accuracy of localization, and unpleasantness. This study was conducted to identify the brain regions associated with the affective-motivational component of tonic skin and muscle pain. METHODS: Forty healthy volunteers were investigated in three groups with different F-18 fluorodeoxyglucose PET activation scans. A verbal rating scale (VRS) was used to quantify pain intensity and unpleasantness. One group was investigated during painful infusion of an acidified phosphate buffer (pH 5.2) into either muscle or skin for 30 minutes. Muscle and skin infusions were adjusted to achieve pain intensity rating of VRS = 40. The second group received sham stimulation of muscle and skin by infusion of non-acidified phosphate buffer (pH 7.3 to 7.4, pain intensity = 0). The third group underwent only one PET scan without sensory stimulation. RESULTS: Unpleasantness ratings were higher (VRS 38.3 vs 25.5) during IM compared to intracutaneous stimulation, despite the same pain intensity (VRS = 40). Sham stimulation revealed no pain or unpleasantness. Regional cerebral glucose metabolism during sham stimulation showed similar findings for intracutaneous and IM infusions with significant activations of the bilateral anterior cingulate, bilateral frontal (premotor) cortex, and the ipsilateral parietal operculum. The comparison of pain vs sham stimulation revealed activations of the bilateral insula for IM but not intracutaneous stimulation. The unpleasantness perception in skin and muscle stimulation was positively correlated to the bilateral insular metabolism. CONCLUSION: The data suggest that the insula represents one main structure where the unpleasantness of tonic pain perception is encoded. PMID- 15824344 TI - Neuropsychological outcome after chemotherapy for primary CNS lymphoma: a prospective study. AB - BACKGROUND: Combined radio- and chemotherapy for primary CNS lymphoma (PCNSL) is associated with a considerable risk of long-term neurotoxicity. The impact of high-dose methotrexate (MTX)-based chemotherapy alone on cognition and quality of life (QOL) is controversial. OBJECTIVE: To assess the impact of the tumor itself and its treatment with high-dose MTX-based chemotherapy on long-term cognition and QOL in patients with PCNSL. METHODS: Prospective neuropsychological examinations and MRI were performed in patients with PCNSL who were in complete remission for more than 12 months after completion of chemotherapy. A QOL assessment was performed at long-term follow-up. RESULTS: Twenty-three patients were eligible. The median follow-up period was 44 months after diagnosis. In long term follow-up, 22 (95%) of 23 patients showed either preserved or improved cognitive functions as compared with pretreatment and immediate posttreatment baseline assessment. One patient showed an isolated decline in psychomotor speed. Eleven (48%) of 23 patients displayed at least mild cognitive deficits at long term follow-up not related to therapy. Nineteen (83%) of 23 patients reported a good QOL. MRI revealed confluent white matter abnormalities in eight patients that were not associated with cognitive decline. CONCLUSION: In patients with primary CNS lymphoma (PCNSL) treated with a methotrexate (MTX)-based chemotherapy, no gross cognitive decline has to be expected as a long-term treatment effect. MTX-induced white matter changes apparent on MRI are not inevitably associated with cognitive impairment. Nevertheless, a substantial fraction of patients with PCNSL retain cognitive deficits as a residual symptom of the tumor. PMID- 15824345 TI - Cellular telephones and risk for brain tumors: a population-based, incident case control study. AB - OBJECTIVE: To evaluate a possible association of glioma or meningioma with use of cellular telephones, using a nationwide population-based case-control study of incident cases of meningioma and glioma. METHODS: The authors ascertained all incident cases of glioma and meningioma diagnosed in Denmark between September 1, 2000, and August 31, 2002. They enrolled 252 persons with glioma and 175 persons with meningioma aged 20 to 69. The authors also enrolled 822 randomly sampled, population-based controls matched for age and sex. Information was obtained from personal interviews, medical records containing diagnoses, and the results of radiologic examinations. For a small number of cases and controls, the authors obtained the numbers of incoming and outgoing calls. They evaluated the memory of the respondents with the Mini-Mental State Examination and obtained data on socioeconomic factors from Statistics Denmark. RESULTS: There were no material socioeconomic differences between cases and controls or participants and non participants. Use of cellular telephone was associated with a low risk for high grade glioma (OR, 0.58; 95% CI, 0.37 to 0.90). The risk estimates were closer to unity for low-grade glioma (1.08; 0.58 to 2.00) and meningioma (1.00; 0.54 to 1.28). CONCLUSION: The results do not support an association between use of cellular telephones and risk for glioma or meningioma. PMID- 15824346 TI - Neurologic features of horizontal gaze palsy and progressive scoliosis with mutations in ROBO3. AB - OBJECTIVE: To review the neurologic, neuroradiologic, and electrophysiologic features of autosomal recessive horizontal gaze palsy and progressive scoliosis (HGPPS), a syndrome caused by mutation of the ROBO3 gene on chromosome 11 and associated with defective decussation of certain brainstem neuronal systems. METHODS: The authors examined 11 individuals with HGPPS from five genotyped families with HGPPS. Eight individuals had brain MRI, and six had electrophysiologic studies. RESULTS: Horizontal gaze palsy was fully penetrant, present at birth, and total or almost total in all affected individuals. Convergence, ocular alignment, congenital nystagmus, and vertical smooth pursuit defects were variable between individuals. All patients developed progressive scoliosis during early childhood. All appropriately studied patients had hypoplasia of the pons and cerebellar peduncles with both anterior and posterior midline clefts of the pons and medulla and electrophysiologic evidence of ipsilateral corticospinal and dorsal column-medial lemniscus tract innervation. Heterozygotes were unaffected. CONCLUSIONS: The major clinical characteristics of horizontal gaze palsy and progressive scoliosis were congenital horizontal gaze palsy and progressive scoliosis with some variability in both ocular motility and degree of scoliosis. The syndrome also includes a distinctive brainstem malformation and defective crossing of some brainstem neuronal pathways. PMID- 15824347 TI - Autosomal recessive mitochondrial ataxic syndrome due to mitochondrial polymerase gamma mutations. AB - OBJECTIVE: To investigate three families and one sporadic case with a recessively inherited ataxic syndrome. METHODS: Clinical and genetic studies were performed in six individuals. Southern blotting and real time PCR were used to detect deletions of mtDNA and mutations in the POLG gene were identified using a combination of DHPLC and direct DNA sequencing. RESULTS: The patients have a distinctive, progressive disorder that starts with episodic symptoms such as migraine-like headache or epilepsy. Ataxia, which is a combination of central and peripheral disease, develops later as does ophthalmoplegia. The commonest form of epilepsy was focal and involved the occipital lobes. Myoclonus was common and patients have a high risk of status epilepticus. MRI typically shows signal changes in the central cerebellum, olivary nuclei, occipital cortex, and thalami. COX negative muscle fibers were found in four of six; in one patient these were rare and in another absent. Multiple mtDNA deletions were identified in all patients, although in two these were not apparent on Southern blotting and real time PCR was required to demonstrate the defect. Two families were homozygous for a previously described POLG mutation, G1399A (A467T). One family and the sporadic case had the same two new mutations, a G to C at position 1491 (Q497H) and a G to C at 2243 (W748S). CONCLUSIONS: Mutations in POLG cause a recessively inherited syndrome with episodic features and progressive ataxia. Characteristic changes on MRI are seen and although skeletal muscle may appear morphologically normal, multiple mtDNA deletions can be detected using real-time PCR. PMID- 15824348 TI - Reliability and validity of the CMT neuropathy score as a measure of disability. AB - OBJECTIVE: To determine the validity and reliability of the Charcot-Marie-Tooth disease (CMT) neuropathy score (CMTNS) in patients with inherited neuropathy. BACKGROUND: Natural history studies and potential treatment trials for patients with various forms of CMT are limited by the lack of quantitative methodologies to monitor disease progression. Most cases of CMT can be considered length dependent axonal neuropathies because disability for even the demyelinating forms correlates with length-dependent axonal degeneration. The total neuropathy score (TNS) is a validated composite measure of disability in length-dependent axonal neuropathies but is weighted toward predominantly sensory neuropathies. Thus, the authors have devised a CMTNS, modified from the TNS, to provide a single measure to quantify CMT disability. METHODS: The authors measured inter- and intrainvestigator reliability of the CMTNS and performed a validation of the score with the Neuropathy Impairment Score (NIS), patient self-assessment scores, an ambulation index, and other measures of disability. RESULTS: Inter- and intrainvestigator reliability was more than 95% in the 60 patients evaluated. Patients could be divided into mild (CMTNS, < or =10), moderate (CMTNS, 11 to 20), and severe (CMTNS, > or =21) categories and demonstrated excellent correlations among all measures of disability. CONCLUSION: The Charcot-Marie Tooth disease (CMT) neuropathy score is a validated measure of length-dependent axonal and demyelinating CMT disability and can be investigated as an end point for longitudinal studies and clinical trials of CMT. PMID- 15824349 TI - Temporal and geographic variation in United States motor neuron disease mortality, 1969-1998. AB - OBJECTIVE: To describe temporal trends of motor neuron disease (MND) mortality in the United States. Variations in MND by demographic variables of sex, age, geography, and race/ethnicity were evaluated to assess the possible explanations for observed trends. METHODS: Multiple-cause mortality files from the National Center for Health Statistics for the years 1969 through 1998 were searched for all United States death records with codes corresponding to MND. Age-adjusted mortality rates were calculated by sex, race/ethnicity, age, birth cohort, and place of death. RESULTS: Overall MND mortality rates increased from 1.25 per 100,000 to 1.82 per 100,000, representing a 46% increase during the 30-year period. Rates among women increased by 60% and continue to rise. Rates among men rose by 35% during this period but have leveled off in the most recent decade evaluated. Mortality rates among African Americans and Hispanics were approximately 50% lower than rates among non-Hispanic whites. A southeast to northwest gradient was observed when rates were grouped by 12 geographic areas. MND mortality rates per 100,000 (and 95% CI) ranged from 2.22 (1.89 to 2.55) in the Northwest to 1.57 (1.44 to 1.71) in the Southeast. CONCLUSIONS: Variations in motor neuron disease (MND) mortality by time, race/ethnicity, sex, and geography were consistent with the hypothesis that environmental exposures, combined with factors of genetic susceptibility, play a role in the development of MND. PMID- 15824350 TI - Cognitive change in ALS: a prospective study. AB - OBJECTIVES: To investigate longitudinally the profile of cognitive impairment in nondemented patients with ALS. METHODS: Twenty nondemented patients with ALS and 18 controls were interviewed at two time points separated by a 6-month interval. The extensive battery was designed to accommodate the range of physical disability present in ALS, and included measures of executive, memory, language, and visuospatial functions, everyday behavior, and emotion. RESULTS: On a test of simple word retrieval (Computerised Sentence Completion Test) patients with ALS became slower over time, while controls became faster at completing sentences with appropriate words. This effect remained when the analysis accommodated for progressive speech disability. Patients with ALS also displayed an impairment in both written and spoken verbal fluency indices (time to think of each word) at both time points, but there was no evidence of deterioration over time. On the Short Inventory of Minor Lapses, carers of patients with ALS displayed increased awareness of cognitive dysfunction in everyday behavior while controls' ratings of their partners decreased. In addition, patients displayed more depressive symptomology (although well below clinical levels) on the second interview vs controls. Patients with ALS also displayed emotional lability at both time points, although this did not increase over time. CONCLUSIONS: Cognitive deterioration in nondemented patients with ALS is a relatively slow process. Selective cognitive impairment in the form of verbal fluency deficits, most likely indicating executive dysfunction, appears relatively early on in the course of the disease, although language functions may become vulnerable as the disease progresses. PMID- 15824351 TI - Pure midbrain infarction: clinical, radiologic, and pathophysiologic findings. AB - OBJECTIVE: To describe the clinical features, MRI findings, and the pathogenesis of the pure midbrain infarction. METHODS: Forty patients with infarcts limited to the midbrain were studied. MRI and angiography (mostly MR angiography) were performed in all patients. RESULTS: Clinical manifestations included gait ataxia in 27 (68%) patients, dysarthria in 22 (55%), limb ataxia in 20 (50%), sensory symptoms in 17 (43%), third nerve palsy in 14 (35%), definitive limb weakness (< or =IV/V) in nine (23%), and internuclear ophthalmoplegia in five (13%). According to MRI findings, the lesions were categorized into four groups. The anteromedial group (n = 18) was characterized by oculomotor disturbances (89%), ataxia (89%, bilateral in 17%), and sensory changes (39%) usually restricted to the perioral and hand areas. Lesions restricted to the subcortical area (n = 10) were usually related to small vessel disease (SVD) (78%), whereas those involving the medial surface (n = 8) were caused by large vessel disease (LVD) (78%). The anterolateral group (n = 11) was characterized by ataxia (70%) and definitive hemiparesis (30%) usually caused by LVD (82%). The combined group (n = 6) had frequent oculomotor disturbances (83%), definitive hemiparesis (67%), and ataxia (50%) and was usually associated with LVD (67%). The lateral group (n = 2) was characterized by prominent sensory symptoms. The prognosis was generally good except for one patient with a bilateral lesion. CONCLUSION: Clinical-radiologic correlation study yields four distinct subgroups: anteromedial, anterolateral, combined, and lateral. Large vessel disease and small vessel disease are usual pathogenic mechanisms, whereas cardiogenic embolism is rare. PMID- 15824352 TI - Thrombolysis in a case of lateral medullary syndrome: CT angiographic findings. PMID- 15824353 TI - Role of cerebral amyloid angiopathy in intracerebral hemorrhage in hypertensive patients. AB - OBJECTIVE: To assess the interaction of cerebral amyloid angiopathy (CAA) and arterial hypertension as cofactors for intracerebral hemorrhage (ICH). METHODS: The authors investigated 129 postmortem brains of hypertensive patients with and without ICH. Sixty-four patients had had deep (n = 40) or lobar (n = 24) ICH. Sixty-five patients without ICH served as controls. Established risk factors for ICH (age, gender, severity of hypertension, bleeding disorders, intake of anticoagulants, and chronic alcoholism) were identified from medical records. Four specimens per brain were stained with hematoxylin-eosin and Congo red. The entire ICH cohort and subgroups were compared with controls using single-factor and multiple logistic regression analyses. RESULTS: CAA was found in 15 of 64 subjects (23%) with ICH and in five of 65 controls (8%; p = 0.026). In single factor analysis, CAA was more prevalent in lobar ICH compared with controls (p = 0.007) but not in deep ICH. Poor control of hypertension was more prevalent in the entire ICH group (p = 0.01) and in deep ICH (p = 0.016) but not in lobar ICH. ICH was predictive of the presence of CAA (odds ratio: 5.6, 95% CI: 1.8 to 19.5, p = 0.003), and CAA was more likely to be found in lobar ICH in multivariable adjusted analysis. After adjustment for conventional risk factors, there was a weak association between CAA and deep ICH. CONCLUSION: Cerebral amyloid angiopathy plays a major role in the pathogenesis of intracerebral hemorrhage even in patients with more evident risk factors. PMID- 15824354 TI - APOE influences vasospasm and cognition of noncomatose patients with subarachnoid hemorrhage. AB - OBJECTIVE: To determine the influence of the APOE genotype on functional and cognitive outcome and on the incidence and prognosis of clinical vasospasm (delayed ischemic neurologic deficit [DIND]) in noncomatose patients with aneurysmal subarachnoid hemorrhage (SAH). METHODS: The authors reviewed the data of patients admitted for SAH to the Neurosurgical Departments of the San Gerardo Hospital of Monza (January 1996 to December 2001) and the Ospedali Riuniti of Bergamo (January 2002 to September 2003). The authors considered only noncomatose patients and evaluated outcome by means of the Rankin Disability Index and the Mini-Mental State Examination at least 6 months after the SAH. STATISTICAL ANALYSIS: Uni- and multivariate logistic regression. RESULTS: The authors included 101 patients. They found the epsilon4 allele in 26 patients (25.7%). The presence of the epsilon4 allele negatively affected the overall outcome (functional morbidity or cognitive morbidity, or both) (p = 0.0087) and, particularly, cognitive morbidity (p = 0.0028). Those with an epsilon4 allele had delayed ischemic neurologic deficit DINDs more frequently (p = 0.024) and, in the presence of DIND, they were more likely to show permanent neurologic deficits (p = 0.0051). CONCLUSIONS: ApoE4 negatively affects cognitive morbidity and delayed ischemic neurologic deficit recovery. The presence of an epsilon4 allele increases the risk of delayed ischemic neurologic deficit. PMID- 15824355 TI - A new dominant distal myopathy affecting posterior leg and anterior upper limb muscles. AB - OBJECTIVE: To report a dominant, slowly progressive early onset distal myopathy with sparing of the tibialis anterior. METHODS: Twelve affected and two possibly affected members from an Australian kindred were examined and investigated by EMG, imaging studies, histopathology, and genetic analysis. RESULTS: Affected patients had a slowly progressive condition with symmetric, distal weakness and wasting of the anterior upper and posterior lower limbs, with sparing of tibialis anterior, even in advanced disease. All patients remained ambulant and there was no evidence of cardiac or respiratory muscle involvement. Serum creatine kinase levels were either normal or mildly elevated. Imaging studies showed widespread involvement of the posterior and lateral leg compartments. Proximal muscles were radiologically abnormal only in advanced disease. Muscles that were mildly affected clinically appeared normal on imaging. EMG in nine patients showed widespread myopathic changes. Muscle histopathology in four patients showed either end stage muscle or nonspecific myopathic findings without inflammation or vacuoles. Microsatellite markers for distal myopathy loci were analyzed and all known distal myopathy phenotype genes and linkage regions were formally excluded by multipoint analysis. CONCLUSIONS: The affected patients in this kindred display a clinically distinct myopathy, with selective involvement of posterior lower and anterior upper limb muscles. The genetic analysis suggests the existence of one more distal myopathy locus. PMID- 15824356 TI - Glycine decarboxylase mutations: a distinctive phenotype of nonketotic hyperglycinemia in adults. AB - Three unrelated adult patients with mild hyperglycinemia, infantile hypotonia, mental retardation, behavioral hyperirritability, and aggressive outbursts were screened for glycine decarboxylase (GLDC) mutations; two novel missense mutations (A389V and R739H) were found. Both mutations had a 6 to 8% of normal GLDC activities when expressed in COS7 cells. PMID- 15824357 TI - The clinical and genetic spectrum of spinocerebellar ataxia 14. AB - Spinocerebellar ataxia 14 (SCA14) is associated with missense mutations in the protein kinase C gamma gene (PRKCG), rather than a nucleotide repeat expansion. In this large-scale study of PRKCG in patients with ataxia, two new missense mutations, an in-frame deletion, and a possible splice site mutation were found and can now be added to the four previously described missense mutations. The genotype/phenotype correlations in these families are described. PMID- 15824358 TI - Measuring Friedreich ataxia: Interrater reliability of a neurologic rating scale. AB - Measuring the severity of neurologic dysfunction in patients with inherited ataxias, including Friedreich ataxia (FA), is difficult because of the variable rate of progression, the variable age at onset and the variety of neural systems that may be affected. The authors discuss the problems related to rating scales in the ataxias, report a neurologic rating scale for FA, and demonstrate acceptable interrater reliability of the instrument. PMID- 15824359 TI - Intrinsic epileptogenicity in polymicrogyric cortex suggested by EEG-fMRI BOLD responses. AB - Polymicrogyria (PMG) is a widespread cortical malformation frequently associated with seizures and EEG spikes. Its epileptogenicity is poorly understood. Nine patients with simultaneous EEG and fMRI were studied to assess the blood oxygenation level-dependent response to spikes. Sixteen of 18 studies showed responses, with maximum activation involving the lesion in 61.5%, but often limited to a small fraction of that lesion, suggesting intrinsic epileptogenicity in small areas of the PMG cortex. PMID- 15824360 TI - Ethmoid abscess with findings simulating Weber syndrome. PMID- 15824361 TI - A randomized controlled crossover trial of aspirin for fatigue in multiple sclerosis. AB - Pharmacotherapeutic options for multiple sclerosis (MS)-related fatigue are limited. Thirty patients were randomly assigned to aspirin (ASA) 1,300 mg/day or placebo in a double-blind crossover study. Results favored ASA for the main clinical outcomes: Modified Fatigue Impact Scale scores (p = 0.043) and treatment preference (p = 0.012). There were no significant adverse effects. The results warrant further study and support a role for ASA-influenced mechanisms, perhaps immunologic, in the generation of MS-related chronic fatigue. PMID- 15824362 TI - An open label study of the effects of rituximab in neuromyelitis optica. AB - Eight patients with worsening neuromyelitis optica were treated with rituximab to achieve B cell depletion. Treatment was well tolerated. Six of eight patients were relapse free and median attack rate declined from 2.6 attacks/patient/year to 0 attacks/patient/year (p = 0.0078). Seven of eight patients experienced substantial recovery of neurologic function over 1 year of average follow-up. The pretreatment median Expanded Disability Status Scale score was 7.5, and at follow up examination was 5.5 (p = 0.013). PMID- 15824363 TI - Encephalopathy and peripheral neuropathy following diethylene glycol ingestion. AB - The authors report a 24-year-old man who developed encephalopathy and rapid quadriplegia following ingestion of a solution containing diethylene glycol (DEG). As quadriparesis evolved, motor response amplitudes were markedly reduced with preserved conduction velocities. Studies during clinical recovery revealed marked motor conduction velocity slowing and prolonged distal latencies. These data indicate that DEG intoxication may cause a primary acute axonal sensorimotor polyneuropathy with demyelinating physiology during recovery. PMID- 15824364 TI - Gadolinium encephalopathy in a patient with renal failure. AB - Gadolinium chelates are extensively used in MRI studies. Neurotoxicity due to gadolinium chelates is minimal and uncommon. A 57-year-old woman in renal failure developed a subacute encephalopathy after inadvertent repetitive gadolinium contrast administration. An unusual MRI appearance with CSF hyperintensity due to gadolinium diffusion into the CSF is also shown. PMID- 15824366 TI - Cranial MRI predicts outcome of spontaneous intracranial hypotension. AB - The outcome of spontaneous intracranial hypotension has been unpredictable. The results of initial MRI were correlated to outcome of treatment in 33 patients with spontaneous intracranial hypotension. A good outcome was obtained in 25 (97%) of 26 patients with an abnormal MRI vs only 1 (14%) of 7 patients with a normal MRI (p = 0.00004). These findings show that normal initial MRI is predictive of poor outcome in spontaneous intracranial hypotension. PMID- 15824367 TI - Central paroxysmal positional vertigo. PMID- 15824365 TI - Methyl bromide intoxication causes reversible symmetric brainstem and cerebellar MRI lesions. AB - Methyl bromide is toxic to the central and peripheral nervous systems. A patient with occupational exposure to this agent is described. MRI showed strikingly symmetric brainstem and cerebellar lesions. The patient's clinical course and the topography and resolution of his MRI abnormalities suggest that this condition is an energy deprivation syndrome. PMID- 15824368 TI - Heparin-induced thrombocytopenia in neurologic patients treated with low molecular-weight heparin. AB - The authors prospectively studied the risk for immune-mediated heparin-induced thrombocytopenia (HIT) in neurologic patients during administration of low molecular-weight heparin (LMWH) vs unfractionated heparin (UFH). None of 111 neurologic patients receiving LMWH developed HIT, whereas HIT occurred in 2.5% of 200 patients treated with UFH (p = 0.17). The rate of heparin-induced antibodies in patients treated with LMWH was lower than in patients treated with UFH (1.8 vs 20.5%; p < 0.001). PMID- 15824369 TI - Microbleeds in hereditary cerebral hemorrhage with amyloidosis-Dutch type. AB - In a hereditary variant of cerebral amyloid angiopathy (CAA), cerebral hemorrhage with amyloidosis-Dutch type, supratentorial microbleeds were found to occur independently of the presence of hypertension, whereas hypertension probably contributed to the development of cerebellar microbleeds. This predictable hereditary variant of CAA may be a useful model to study microbleeds in relation to CAA. PMID- 15824371 TI - CSF Abeta42 and tau or phosphorylated tau and prediction of progressive mild cognitive impairment. AB - Baseline CSF amyloid beta-peptide-42 (Abeta42), tau, and phosphorylated tau (P tau) levels from 46 control subjects and 78 patients with mild cognitive impairment (MCI) were measured. Twenty-three patients with MCI developed dementia during the study. Abnormal biomarkers were found early in the course of Alzheimer disease (AD). The most predictive assay for AD among the patients with MCI was the combination of Abeta42 and P-tau. PMID- 15824370 TI - Sequential antibodies to potassium channels and glutamic acid decarboxylase in neuromyotonia. AB - A patient with thymoma-associated neuromyotonia and voltage-gated potassium channel (Kv1.2 and Kv1.6) antibodies by immunoprecipitation and rat brain immunolabeling was treated successfully with immunoadsorption and cyclophosphamide. Curiously, glutamic acid decarboxylase antibodies, absent at onset, appeared later. Stiff-person syndrome was absent, but fast blink reflex recovery suggested enhanced brainstem excitability. The range of antibodies produced in thymoma-associated neuromyotonia is richer, and the timing of antibody appearance more complex, than previously suspected. PMID- 15824372 TI - A pilot, double-blind, placebo-controlled trial of indinavir in patients with ALS. AB - There is some evidence of retroviral infection in ALS. A randomized, double blind, placebo-controlled trial of indinavir in ALS was performed to assess safety and efficacy trends. Nephrolithiasis and gastrointestinal side effects were frequent with indinavir treatment. Group differences in the rate of decline were not significant between the groups for the ALS Functional Rating Scale (p = 0.36) or for the secondary variables. The toxicity and negative efficacy trends discourage further indinavir trials in ALS. PMID- 15824373 TI - Smoking affects the phenotype of Alzheimer disease. AB - Because epidemiologic and in vitro evidence conflict, the authors studied the association between smoking and Alzheimer disease (AD) in 46 never, 47 former, and 15 active smokers with AD followed to autopsy. Disease parameters were examined by smoking status and amount smoked in bivariate tests and in multivariate models controlling for age, sex, education, and APOE status. Smoking status was not associated with cognitive or neuropathologic measures. However, active smokers were significantly younger at death and higher levels of smoking were associated with shorter disease duration. PMID- 15824375 TI - Assessment of cognitive function and communication ability in a completely locked in patient. PMID- 15824374 TI - PRNP H187R mutation associated with neuropsychiatric disorders in childhood and dementia. AB - Described is a large family with an autosomal dominant dementia associated with an H187R mutation in the prion protein gene (PRNP). Clinical features include neuropsychiatric disturbances in childhood and adolescence, dementia in young adulthood with frontotemporal manifestations, and long disease duration. Neuropathology revealed atrophy and mild gliosis, whereas prion protein analysis revealed an abnormal conformer with unusual sensitivity to protease digestion. Mutations in PRNP may cause neuropsychiatric disorders that predate dementia by many years. PMID- 15824376 TI - Headache in cranial and cervical dystonia. PMID- 15824377 TI - Amygdala dysplasia with temporal lobe epilepsy and obsessive-compulsive disorder: an fMRI/EEG study. PMID- 15824379 TI - Jerky seesaw nystagmus in isolated internuclear ophthalmoplegia from focal pontine lesion. PMID- 15824378 TI - Nonepileptic visual hallucinations in Lafora disease. PMID- 15824380 TI - Personal history: resolution. PMID- 15824381 TI - Recombinant hepatitis B vaccine and the risk of multiple sclerosis: a prospective study. PMID- 15824382 TI - Season of birth and risk of brain tumors in adults. PMID- 15824383 TI - Sporadic Creutzfeldt-Jakob disease: magnetic resonance imaging and clinical findings. PMID- 15824384 TI - Statin therapy and risk of dementia in the elderly. PMID- 15824385 TI - Cosmetic neurology: the controversy over enhancing movement, mentation, and mood. PMID- 15824386 TI - Economic evaluation of donepezil in moderate to severe Alzheimer disease. PMID- 15824387 TI - Avian influenzavirus: are we prepared? PMID- 15824389 TI - The ethics of research involving Canada's Aboriginal populations. PMID- 15824391 TI - Clinical trials registry. PMID- 15824393 TI - Clinical trials registry. PMID- 15824394 TI - Clinical trials registry. PMID- 15824395 TI - Debate over credibility of natural health product claims. PMID- 15824396 TI - Russia underestimates HIV/AIDS incidence. PMID- 15824397 TI - More resident positions needed say medical students. PMID- 15824398 TI - Dangerous copyright proposals hit medical community hard. PMID- 15824401 TI - Canadian guidelines for body weight classification in adults: application in clinical practice to screen for overweight and obesity and to assess disease risk. PMID- 15824402 TI - Hand-arm vibration syndrome. PMID- 15824403 TI - Alkaptonuria and photography: a patient's urine tells the story. PMID- 15824404 TI - Is there a role for estrogen in the prevention and treatment of urinary incontinence? PMID- 15824405 TI - Epidemiology of severe trauma among status Aboriginal Canadians: a population based study. AB - BACKGROUND: Aboriginal Canadians are considered to be at increased risk of major trauma. However, population-based studies characterizing the distribution, determinants and outcomes of major trauma in this group are lacking. We sought to measure the impact of ethnicity, as reflected by Aboriginal status, on the incidence of severe trauma and to broadly define the epidemiologic characteristics of severe trauma among status Aboriginal Canadians in a large health region. METHODS: This population-based, observational study involves all adults (people > or = 16 years) resident in the Calgary Health Region between Apr. 1, 1999, and Mar. 31, 2002. Stratification of the population into status Aboriginal Canadians and the reference population was performed by Alberta Health and Wellness using an alternate premium arrangement field within the personal health care number. Injury incidence was determined by identifying all injuries with severity scores of 12 or greater in the Alberta Trauma Registry, regional corporate data and the Office of the Medical Examiner. RESULTS: Aboriginal Canadians were at much higher risk than the reference population in the Calgary Health Region of sustaining severe trauma (257.2 v. 68.8 per 100,000; relative risk [RR] 3.7, 95% confidence interval [CI] 3.0-4.6). Aboriginal Canadians were found to be at significantly increased risk of injuries resulting from motor vehicle crashes (RR 4.8, 95% CI 3.5-6.5), assault (RR 11.1, 95% CI 6.2-18.6) and traumatic suicide (RR 3.1, 95% CI 1.4-6.1). A trend toward higher median injury severity scores was observed among Aboriginal Canadians (21 v. 18, p = 0.09). Although the case-fatality rate among Aboriginal Canadians was less than half that in the reference population (14/93 [15%] v. 531/1686 [31%], p < 0.0001), population mortality was almost 2 times greater (RR = 1.8, 95% CI 1.0-3.0, p = 0.046). INTERPRETATION: Severe trauma disproportionately affects Aboriginal Canadians. PMID- 15824406 TI - Rate of elective cholecystectomy and the incidence of severe gallstone disease. AB - BACKGROUND: The use of elective cholecystectomy has increased dramatically following the widespread adoption of laparoscopic cholecystectomy. We sought to determine whether this increase has resulted in a reduction in the incidence of severe complications of gallstone disease. METHODS: We examined longitudinal trends in the population-based rates of severe gallstone disease from 1988 to 2000, using a quasi-experimental longitudinal design to assess the effects of the large increase in elective cholecystectomy rates after 1991 among people aged 18 years and older residing in Ontario. We also measured the rate of hospital admission because of acute diverticulitis, to control for secular trends in the use of hospital care for acute abdominal diseases. RESULTS: The adjusted annual rate of elective cholecystectomy per 100,000 population increased from 201.3 (95% confidence interval [CI] 197.0-205.8) in 1988-1990 to 260.8 (95% CI 257.1- 264.5) in 1992-2000 (rate ratio [RR] 1.35, 95% CI 1.32- 1.38, p 0.001). An anomalously high number of elective cholecystectomies were performed in 1991. Overall, the annual rate of severe gallstone diseases (acute cholecystitis, acute biliary pancreatitis and acute cholangitis) declined by 10% (RR 0.90, 95% CI 0.88- 0.91) for 1992-2000 as compared with 1988-1991. This decline was entirely due to an 18% reduction in the rate of acute cholecystitis (RR 0.82, 95% CI 0.80-0.84). INTERPRETATION: The increase in the rate of elective cholecystectomy that occurred following the introduction of laparoscopic cholecystectomy in 1991 was associated with an overall reduction in the incidence of severe gallstone disease that was entirely attributable to a reduction in the incidence of acute cholecystitis. PMID- 15824407 TI - Getting to the root of trauma in Canada's Aboriginal population. PMID- 15824408 TI - A call to bloggers (not a running group). PMID- 15824409 TI - The 2003 Canadian recommendations for dyslipidemia management: revisions are needed. PMID- 15824410 TI - The analysis by Manuel and colleagues creates controversy with headlines, not data. PMID- 15824412 TI - Intra-articular hyaluronic acid for the treatment of osteoarthritis of the knee: systematic review and meta-analysis. AB - BACKGROUND: Osteoarthritis of the knee affects up to 10% of the elderly population. The condition is frequently treated by intra-articular injection of hyaluronic acid. We performed a systematic review and meta-analysis of randomized controlled trials to assess the effectiveness of this treatment. METHODS: We searched MEDLINE, EMBASE, CINAHL, BIOSIS and the Cochrane Controlled Trial Register from inception until April 2004 using a combination of search terms for knee osteoarthritis and hyaluronic acid and a filter for randomized controlled trials. We extracted data on pain at rest, pain during or immediately after movement, joint function and adverse events. RESULTS: Twenty-two trials that reported usable quantitative information on any of the predefined end points were identified and included in the systematic review. Even though pain at rest may be improved by hyaluronic acid, the data available from these studies did not allow an appropriate assessment of this end point. Patients who received the intervention experienced a reduction in pain during movement: the mean difference on a 100-mm visual analogue scale was -3.8 mm (95% confidence interval [CI] -9.1 to 1.4 mm) after 2-6 weeks, -4.3 mm (95% CI -7.6 to -0.9 mm) after 10-14 weeks and -7.1 mm (95% CI -11.8 to -2.4 mm) after 22-30 weeks. However, this effect was not compatible with a clinically meaningful difference (expected to be about 15 mm on the visual analogue scale). Furthermore, the effect was exaggerated by trials not reporting an intention-to-treat analysis. No improvement in knee function was observed at any time point. Even so, the effect of hyaluronic acid on knee function was more favourable when allocation was not concealed. Adverse events occurred slightly more often among patients who received the intervention (relative risk 1.08, 95% CI 1.01 to 1.15). Only 4 trials explicitly reported allocation concealment, had blinded outcome assessment and presented intention-to treat data. INTERPRETATION: According to the currently available evidence, intra articular hyaluronic acid has not been proven clinically effective and may be associated with a greater risk of adverse events. Large trials with clinically relevant and uniform end points are necessary to clarify the benefit-risk ratio. PMID- 15824413 TI - Fertility after breast cancer: questions abound. PMID- 15824414 TI - Phase I study of oblimersen sodium, an antisense to Bcl-2, in untreated older patients with acute myeloid leukemia: pharmacokinetics, pharmacodynamics, and clinical activity. AB - PURPOSES: Pharmacologic downregulation of Bcl-2, an antiapoptotic protein overexpressed in cancer, might increase chemosensitivity in acute myeloid leukemia (AML). Herein, we investigated the feasibility of this approach in untreated elderly AML patients by administering oblimersen sodium (G3139), an 18 mer phosphorothioate antisense to Bcl-2, during induction and consolidation treatments. PATIENTS AND METHODS: Untreated patients with primary or secondary AML (stratified to cohort 1 or 2, respectively) who were > or = 60 years received induction with G3139, cytarabine, and daunorubicin at one of two different dose levels (45 and 60 mg/m2) and, on achievement of complete remission (CR), consolidation with G3139 and high-dose cytarabine. An enzyme-linked immunosorbent assay (ELISA)-based assay was used to measure plasma and intracellular concentrations (IC) of G3139. Bcl-2 mRNA and protein levels were quantified by real-time reverse transcriptase polymerase chain reaction and ELISA, respectively, in bone marrow samples collected before induction treatment and after 72 hours of G3139 infusion, prior to initiation of chemotherapy. RESULTS: Of the 29 treated patients, 14 achieved CR. With a median follow-up of 12.6 months, seven patients had relapsed. Side effects of this combination were similar to those expected with chemotherapy alone and were not dose limiting at both dose levels. After 72-hour G3139 infusion, Bcl-2/ABL mRNA copies were decreased compared with baseline (P = .03) in CR patients and increased in nonresponders (NRs; P = .05). Changes in Bcl-2 protein showed a similar trend. Although plasma pharmacokinetics did not correlate with disease response, the median IC of the antisense was higher in the CR patients compared with NRs (17.0 v 4.4 pmol/mg protein, respectively; P = .05). CONCLUSION: G3139 can be administered safely in combination with intensive chemotherapy, and the degree of Bcl-2 downmodulation may correlate with response to therapy. PMID- 15824415 TI - Ablative allogeneic hematopoietic cell transplantation in adults 60 years of age and older. AB - PURPOSE: To evaluate outcomes of ablative allogeneic hematopoietic cell transplantation (HCT) in older patients with hematologic malignancies. PATIENTS AND METHODS: We treated 52 patients from 1979 to 2002 with a median age of 62.8 years (range, 60.1 to 67.8 years) using ablative preparative regimens followed by allogeneic HCT from sibling donors. Diagnoses included myelodysplastic syndrome (MDS; n = 35), chronic myeloid leukemia (CML; n = 8), acute myeloid leukemia (AML; n = 6), and other (n = 3). Conditioning regimens included cyclophosphamide (CY) and busulfan (BU) (67%), total-body irradiation and CY (21%), BU-fludarabine (10%), and CY (2%). RESULTS: Eighteen (35%) of 52 patients are alive at a median of 4.6 years (range, 0.8 to 9.1 years) after transplantation. Median overall survival (OS) and progression-free survival were 300 and 218 days, respectively. Three-year OS and relapse rates are estimated to be 34% and 24%, respectively. Nonrelapse mortality (NRM) rates at 100 days and 3 years are estimated to be 27% and 43%, respectively. Grade 3 to 4 acute graft-versus-host disease (GVHD) occurred in 20% of patients, and chronic extensive GVHD was described in 53% of patients. Fourteen (40%) of 35 patients with MDS are alive at a median of 2.8 years (range, 0.8 to 8.2 years). Four of six patients with CML in chronic or accelerated phase are alive at a median of 6.9 years (range, 4.1 to 9.1 years) after transplantation. None of the patients with AML, CML in blast crisis, or other diagnoses have survived. Patients who underwent transplantation after 1993 had improved survival. CONCLUSION: These data suggest that allogeneic HCT is feasible in selected patients > or = 60 years of age, although novel methods to reduce NRM while maintaining efficacy are needed. PMID- 15824416 TI - Fertility preservation in breast cancer patients: a prospective controlled comparison of ovarian stimulation with tamoxifen and letrozole for embryo cryopreservation. AB - PURPOSE: To develop safe ovarian stimulation methods to perform in vitro fertilization (IVF) in breast cancer patients who wish to preserve their fertility via embryo cryopreservation before chemotherapy. PATIENTS AND METHODS: Sixty women (age range, 24 to 43 years) with breast cancer were prospectively studied. Twenty-nine patients underwent 33 ovarian stimulation cycles with either tamoxifen 60 mg/d alone (Tam-IVF) or in combination with low-dose follicle stimulating hormone (TamFSH-IVF) or letrozole 5 mg in combination with FSH (Letrozole-IVF). After IVF, all resultant embryos were cryopreserved to preserve fertility. Recurrence rates were compared with controls (n = 31) who elected not to undergo IVF. RESULTS: Compared with Tam-IVF, both TamFSH-IVF and Letrozole-IVF patients had greater numbers of follicles (2 +/- 0.3 v 6 +/- 1 and 7.8 +/- 0.9, respectively; P < .0001), mature oocytes (1.5 +/- 0.3 v 5.1 +/- 1.1 and 8.5 +/- 1.6, respectively; P < .001), and embryos (1.3 +/- 0.2 v 3.8 +/- 0.8 and 5.3 +/- 0.8, respectively; P < .001). Peak estradiol (E2) levels were lower with Letrozole-IVF and Tam-IVF compared with TamFSH-IVF. After 554 +/- 31 days (range, 153 to 1,441 days) of follow-up, cancer recurrence rate was similar between IVF and control patients (three of 29 v three of 31 patients, respectively; hazard ratio, 1.5; 95% CI, 0.29 to 7.4), and this estimate was not affected by cancer stage. CONCLUSION: The combination of low-dose FSH with tamoxifen (TamFSH-IVF) or letrozole (Letrozole-IVF) results in higher embryo yield compared with Tam-IVF. Recurrence rates do not seem to be increased, but the letrozole protocol may be preferred because it results in lower peak E2 levels. PMID- 15824418 TI - Identification of two new Helicobacter pylori surface proteins involved in attachment to epithelial cell lines. AB - Helicobacter pylori causes the development of gastritis, gastric ulcers and adenocarcinomas in humans. The establishment of infection is influenced by adherence to the gastric epithelium, and several bacterial adhesins and host cell receptors have been identified. H. pylori recognize the Lewis(b) receptor through the BabA adhesin but also readily adhere to epithelia in the absence of the Lewis(b) epitope, demonstrating the relevance of additional adhesive interactions. This study presents a novel method of identifying bacterial adhesins. Nickel beads were coated with H. pylori-derived, recombinantly expressed ORF proteins, and epithelial cells from the human stomach, intestine or urinary tract were allowed to adhere to those beads. The binding of epithelial cells to the protein-coated nickel beads was confirmed by electron microscopy or flow cytometry using antibodies directed towards the His-tags. Among the five ORFs tested, two new adhesive proteins (HP1188 and HP1430) were identified. Both were expressed on the surface of virulent H. pylori, with the HP1188 protein being most abundant. The purified HP1188 and HP1430 proteins bound more strongly to gastric than to other epithelial cell lines, suggesting that they may be involved in the colonization of the human gastric mucosa. In conclusion, this method facilitates the identification of ORFs of microbial origin involved in cellular interactions such as adherence. PMID- 15824419 TI - Antigenic and phenotypic modifications of Yersinia pestis under calcium and glucose concentrations simulating the mammalian bloodstream environment. AB - To study the possible mechanism of extracellular resistance to phagocytes developed by Yersinia pestis in the early stage of plague infection, the behaviour of two Y. pestis strains, the vaccine EV-76 and fully virulent 231 (LD(50), 10 c.f.u.), was studied in-depth after cultivation in vitro at the host temperature in conditions simulating the bloodstream environment of mammals. For this, two standard basal media supplemented with calcium and glucose in appropriate concentrations were employed: Hottinger broth, routinely used for growth of Y. pestis in vitro, and RPMI 1640, simulating human extracellular fluid. Although both media permitted Y. pestis to achieve the resistant state, RPMI enabled significantly higher bacterial proliferation and increased modifications in the production of the principal surface antigens that affect the relevant phenotype characteristics. In general, our results indicate that the Y. pestis bacteria in the resistant state do not produce species-specific antigens, i.e. fraction 1 or F1, 'murine' toxin or Ymt, plasminogen activator (Pla) and any surface-specific polysaccharides, resulting in unmasking of the cross-reactive epitopes of lipid A in reduced Y. pestis lipopolysaccharide. This may produce mimicry by Y. pestis of some human tissue and blood cell components, with no immune response and inflammation at the site of infection at the early stage, which enables Y. pestis to survive, extensively multiply and spread into the circulation. PMID- 15824420 TI - Immunogenicity of mycobacterial PPE44 (Rv2770c) in Mycobacterium bovis BCG infected mice. AB - The Pro-Pro-Glu (PPE) protein family of Mycobacterium tuberculosis includes 69 glycine-rich proteins with a conserved N-terminal domain. Their role in tuberculosis is unknown, but it has been speculated that they may have an important immunological significance. In this investigation, the immunogenicity of the ppe44 (Rv2770c) gene product in BALB/c mice infected subcutaneously or intravenously with Mycobacterium bovis bacille Calmette-Guerin (BCG) was evaluated. Mice infected subcutaneously developed high titres of anti-PPE44 IgG1 antibodies, while PPE44-specific IgG2a antibodies were absent at all times tested. PPE44-primed cells from draining lymph nodes and spleen produced low levels of IFN-gamma, and a moderate degree of delayed-type hypersensitivity was observed following PPE44 intracutaneous challenge. In mice infected intravenously, the anti-PPE44 IgG1 antibody response was markedly higher compared with the subcutaneous infection; anti-PPE44 IgG2a antibodies at titres approximately 0.5-2.0 log(10) lower than IgG1 were detected. Interferon (IFN) gamma production in PPE44-stimulated spleen-cell cultures was transient. These results indicate that PPE44 represents a novel mycobacterial antigen expressed during subcutaneous and intravenous infection by M. bovis BCG in BALB/c mice. Both infection models seem to polarize the immune response to PPE44 towards a Th2 phenotype, as testified by the IgG1 isotype being predominant over IgG2a and by the low IFN-gamma and delayed-type hypersensitivity responses. PMID- 15824421 TI - Sensitivity of laser light depolarization analysis for detection of malaria in blood samples. AB - Automated light depolarization analysis could be a useful tool for diagnosing malarial infections. This work discusses the results of a diagnostic efficacy study on 411 samples from patients with suspected malaria infection performed with a Cell-Dyn 4000 analyser. Light dispersed at 90 degrees and depolarized can be used for identifying and counting eosinophils. However, other cell populations with depolarizing capacity occur in malarial samples; these result from leukocytes ingesting haemozoin that is derived from the degradation of the haem group of haemoglobin performed by the parasite. A sensitivity of 72 % and specificity of 98 % were recorded, with positive and negative predictive values of 78 % and 97 %, respectively. Although the sensitivity level of the automated light depolarization analysis is not adequate to replace the existing methods for the diagnosis of parasitic diseases, it could alert clinicians to unsuspected infections by parasites, particularly those from the genus Plasmodium. PMID- 15824422 TI - Failure to detect capsule gene bexA in Haemophilus influenzae types e and f by real-time PCR due to sequence variation within probe binding sites. AB - Detection of the conserved capsule gene bexA is used to distinguish capsulate from non-capsulate Haemophilus influenzae. While developing a real-time PCR assay to detect bexA, it was found that bexA probes produced a detectable signal for H. influenzae types a to d, but failed to do so for H. influenzae types e and f. Sequencing revealed differences compared with H. influenzae types a to d within probe binding sites. To prevent misclassification of strains as non-capsulate, assays must detect all capsular types. PMID- 15824423 TI - Diagnosis of human brucellosis caused by Brucella canis. AB - The transmission of Brucella canis to man commonly occurs through contact with infected dogs or their secretions, or through direct laboratory exposure. The disease is underdiagnosed due to a general lack of serological testing facilities and misconceptions concerning its prevalence. This report shows the potential use of an indirect ELISA (IELISA) for the diagnosis of human brucellosis caused by B. canis in a population of patients negative by smooth-Brucella antigen tests but positive by rapid slide agglutination test (RSAT). One hundred and ten sera from asymptomatic people found negative by tests using smooth Brucella abortus antigen and by RSAT showed an IELISA specificity of 100 % when a cut-off value of 27 % positivity (%P) was selected. For 17 sera from patients with positive B. canis culture or in close contact with culture-positive dogs, the IELISA sensitivity was 100 % with the same cut-off value. The positive patients presented clinical symptoms similar to brucellosis caused by other species of Brucella and some of them received antibiotic treatment and made good progress. Using this cut-off value, we studied 35 patients with negative blood cultures but positive RSATs, and IELISA detected 18 as positive; of the 17 IELISA-negative, two were RSAT positive at dilution 1 : 2 and 15 were weakly positive with pure serum. These samples were probably from patients at an early stage of infection or indicate false-positive results. No cross-reaction was observed among the sera from nine cases with a diagnosis other than brucellosis, but cross-reactivity was evident in sera from patients infected with smooth-Brucella species. Since routine brucellosis diagnosis does not include B. canis investigation, infection with this species may be more widespread than is currently suspected. The RSAT could be a suitable screening test for the diagnosis of B. canis human brucellosis, and a supplementary technique, such as IELISA, performed on all positive RSAT samples that were negative by B. abortus antigen could ensure diagnostic specificity and confirm the diagnosis. PMID- 15824424 TI - A novel porA-based real-time PCR for detection of meningococcal carriage. AB - Real-time PCR based on the capsule transfer gene (ctrA) is a significant aid in the diagnosis of meningococcal infection but fails to detect a high proportion (60 %) of non-groupable strains associated with nasopharyngeal carriage. This study aimed to design a novel real-time (TaqMan) PCR that would detect more strains of meningococci and be suitable for large-scale carriage studies. Primer and probe sequences were based on the meningococcal porA gene and designed specifically to exclude the highly related porA pseudogene in Neisseria gonorrhoeae. The specificity of the assay was confirmed by testing strains of N. gonorrhoeae known to contain the porA pseudogene together with commensal strains of Neisseria lactamica and Neisseria sicca. None of these was detected in the assay. Neisseria meningitidis strains representing a wide range of serogroups together with non-groupable strains isolated from the nasopharynx were tested by ctrA assay and the novel porA-based TaqMan PCR. All carriage strains were detected by the porA-based assay including four that gave weak or no reaction with the ctrA assay. Comparison of ctrA and porA assays on 71 throat swabs obtained from university students showed that the porA assay detected meningococcal DNA in all samples that were ctrA positive plus three that were ctrA negative but culture positive. This novel porA-based TaqMan assay provides a highly specific method for detecting meningococcal DNA that is more sensitive than the ctrA assay for detecting meningococcal carriage and is particularly suitable for carriage studies where non-groupable strains and other Neisseria are present. PMID- 15824425 TI - Comparison of serotyping, pulsed field gel electrophoresis and amplified fragment length polymorphism for typing of Streptococcus pneumoniae. AB - The aim of the present study was to compare serotyping, PFGE and AFLP for typing of Streptococcus pneumoniae with regard to discriminatory power, typeability and typing system concordance. Thirty-four isolates from cerobrospinal fluid and 34 time-matched blood culture isolates collected from in-patients at two hospitals in western Norway during the period from January 1994 to May 2002 were included in the study. The discriminatory powers of serotyping, PFGE and AFLP were 0.93, 0.99 and 0.95, respectively. The typeabilities for serotyping, PFGE and AFLP were 1, 1 and 0.99, respectively. A good concordance was shown between all the typing methods. Serotyping would most probably have a higher discriminatory power if further subtyping had been performed. PFGE was more discriminatory than AFLP, and AFLP grouped more-distantly related isolates together. The two typing methods thus provided different information, and therefore both could be useful adjuncts to serotyping for the characterization of S. pneumoniae. PMID- 15824426 TI - Spore shedding pattern of Enterocytozoon bieneusi in asymptomatic children. AB - Stool samples from seven human immunodeficiency virus (HIV)-negative and two HIV positive children with asymptomatic Enterocytozoon bieneusi infections were daily examined to quantify spore shedding using Gram-chromotrope staining under light microscopy. The spore shedding pattern and intensity in these children was variable. Mean spore concentrations in the stool samples from these children ranged from 2.4 x 10(2) to 1.2 x 10(5) spores per gram. Light microscopy could detect spores in stool specimens for 9-33 days, while PCR was able to detect E. bieneusi in stool specimens for 3-40 days longer. This suggests that light microscopy may not detect low levels of spore shedding. Considering that the asymptomatic group are a potential source of infection, detection methods with a higher sensitivity should be used. PMID- 15824427 TI - Isolation and characterization of provisional serovar Shigella boydii E16553 from diarrhoeal patients in Bangladesh. AB - In previous studies with strains of the Shigella dysenteriae provisional serovars E22383 and E23507 from diarrhoeal stools from patients in Bangladesh, two strains of Shigella species were identified as Shigella boydii provisional serovar E16553 by a reference laboratory. Further tests with an antiserum to an international type strain of the provisional serovar E16553 identified an additional 15 isolates. None of the isolates reacted with antisera to the established Shigella serovars or any other provisional serovars reported so far and all showed biochemical reactions typical of S. boydii. All of the isolates harboured the 140 MDa invasion plasmid, had the ipaH gene and produced keratoconjunctivitis in the guinea pig eye. All isolates were susceptible to ampicillin, sulfamethoxazole trimethoprim, nalidixic acid, ciprofloxacin and mecillinam but eight strains were resistant to tetracycline. A single PFGE type (type A) was shown for all 17 clinical isolates, indicating a common source of origin. The pulsotype of the Bangladeshi isolates was closely related to that of a Japanese strain but was different from that of the type strain. On the basis of these biochemical, serological and virulence markers, and diverse geographical origin, it is recommended that the provisional status of serovar E16553 be changed and that it be included in the international serotyping classification scheme as S. boydii 19. PMID- 15824428 TI - Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods. AB - Propolis exhibits antimicrobial, anti-inflammatory and other biological effects. The aim of this study was to evaluate the activity of 30 % ethanolic extract of Bulgarian propolis against 94 Helicobacter pylori strains by three methods. By the agar-well diffusion method, only 13.8 % of the strains exhibited no inhibition by 30 microl propolis extract (containing 9 mg propolis) and all isolates were inhibited to some extent by 90 microl of the extract (27 mg propolis) per well. The mean diameters of growth inhibition by 30, 60 or 90 microl propolis extract or 30 microl 96 % ethanol per well were 16.8, 19.2, 27.5 and 8.3 mm, respectively. The propolis extract was more active than the ethanol (P < 0.001). With 90 microl propolis extract per well, 69.4 % of the strains exhibited large diameters of growth inhibition (> or =20 mm) versus 26.6 % with 30 mul per well (P < 0.001). With moist propolis discs, inhibition was detected in more strains (92.1 %) than with dried discs (78.2 %, P < 0.05), with mean inhibitory diameters of 18.7 and 13.8 mm, respectively. By the agar dilution method, 100 and 300 microg propolis ml(-1) inhibited the growth of 57.1 % and 76.2 %, respectively, of the 21 strains tested. In conclusion, Bulgarian propolis had a strong and dose-dependent activity against most of the H. pylori strains tested. Although the effect of propolis on H. pylori in vitro is promising, further microbiological, pharmacological and clinical trials are required. PMID- 15824429 TI - Colonization of 8-week-old conventionally reared goats by Escherichia coli O157 : H7 after oral inoculation. AB - Enterohaemorrhagic Escherichia coli O157 : H7 infections of man have been associated with consumption of unpasteurized goat's milk and direct contact with kid goats on petting farms, yet little is known about colonization of goats with this organism. To assess the contribution of flagella and intimin of E. coli O157 : H7 in colonization of the goat, 8-week-old conventionally reared goats were inoculated orally in separate experiments with 1x10(10) c.f.u. of a non verotoxigenic strain of E. coli O157 : H7 (strain NCTC 12900 Nal(r)), an aflagellate derivative (DMB1) and an intimin-deficient derivative (DMB2). At 24 h after inoculation, the three E. coli O157 : H7 strains were shed at approximately 5x10(4) c.f.u. (g faeces)(-1) from all animals. Significantly fewer intimin deficient bacteria were shed only on days 2 (P = 0.003) and 4 (P = 0.014), whereas from day 7 to 29 there were no differences. Tissues from three animals inoculated with wild-type E. coli O157 : H7 strain NCTC 12900 Nal(r) were sampled at 24, 48 and 96 h after inoculation and the organism was cultured from the large intestine of all three animals and from the duodenum and ileum of the animal examined at 96 h. Tissues were examined histologically but attaching-effacing (AE) lesions were not observed at any intestinal site of the animals examined at 24 or 48 h. However, the animal examined at 96 h, which had uniquely shed approximately 1x10(7) E. coli O157 : H7 (g faeces)(-1) for the preceding 3 days, showed a heavy, diffuse infection with cryptosporidia and abundant, multifocal AE lesions in the distal colon, rectum and at the recto-anal junction. These AE lesions were confirmed by immunohistochemistry to be associated with E. coli O157 : H7. PMID- 15824430 TI - DNA identification of the pathogen of candidal aspiration pneumonia induced in the course of oral cancer therapy. AB - Aspiration of oropharyngeal bacteria and fungi is occasionally suspected in patients with pneumonia. A patient with oral carcinoma underwent chemoradioimmunotherapy and, about 4 weeks from the start of the therapy, the patient suffered from severe oral mucositis induced by chemoradiotherapy, and candidal pneumonia was subsequently induced. The candidal pneumonia was insufficiently improved by potent antifungal drugs, taking a lethal course. Randomly amplified polymorphic DNA analysis and DNA sequence examination of strains isolated from the oral cavity 1 week before the onset of pneumonia and autopsied lung revealed the identity of both strains as Candida albicans, and the DNA analysis supported aspiration of oral Candida. These results indicate that the pathogen of the pneumonia, C. albicans, was aspirated from the oral cavity and that oral Candida is easily aspirated and becomes the pathogen of pneumonia. PMID- 15824431 TI - Concurrence between the gene expression pattern of Actinobacillus actinomycetemcomitans in localized aggressive periodontitis and in human epithelial cells. AB - Actinobacillus actinomycetemcomitans is a facultatively intracellular pathogen and the aetiological agent of localized aggressive periodontitis. Screening of the genome of A. actinomycetemcomitans for in vivo-induced antigen determinants previously demonstrated that the proteome of this organism differs in laboratory culture compared with conditions found during active infection. The aim of the present study was to determine whether the bacterial gene expression pattern inferred with in vivo-induced antigen technology (IVIAT) in human infections was consistent with the gene expression pattern occurring upon epithelial cell association. To this end, a real-time PCR method was developed and used to quantify absolute and relative bacterial gene expression of A. actinomycetemcomitans grown extra- and intracellularly in two human epithelial cell lines (HeLa and IHGK). The amount of template used in the assay was normalized using the total count of viable bacteria (c.f.u.) as a reference point and performed in duplicate in at least two independent experiments. Controls for this experiment included 16S rRNA and gapdh. Transcription of all eight ORFs tested increased significantly (P < 0.05) in HeLa and IHGK cells compared with bacteria grown extracellularly. The concurrence of gene expression patterns found in the two models suggests that these epithelial cells are valid in vitro models of infection for the genes tested. IVIAT is an experimental platform that can be used as a validation tool to assess the reliability of animal and other models of infection and is applicable to most pathogens. PMID- 15824432 TI - Unusual clinical presentation of brucellosis caused by Brucella canis. AB - Brucella canis is considered a rare cause of human brucellosis. The clinical importance of this infection may have been underestimated so far because of difficulties with presumptive diagnosis. The case described here presented symptoms compatible with brucellosis but the routine tests using Brucella abortus antigen were negative. The infection would have remained undiagnosed if culture had not been positive. This case illustrates the potential for a favourable outcome in Brucella canis diagnosis and supports recommendations for the use of B. canis serology. The infection should be suspected in patients with compatible symptoms and negative serology for B. abortus antigen. PMID- 15824433 TI - Evidence for low temperature regulation of biofilm formation in Staphylococcus epidermidis. PMID- 15824434 TI - An unusual cluster of dysentery due to Shigella dysenteriae type 4 in Dhaka, Bangladesh. PMID- 15824435 TI - The century beyond the gene. AB - In an increasing number of biological laboratories, the focus of research is shifting from sequence data to the functional meaning of that data. No longer content with structural mappings, there is a renewed interest abroad in what the United States Department of Energy calls, 'Bringing Genomes to Life'. For many, this means a movement beyond 'reductionism' to a 'systems biology'. The question is, what does this mean? PMID- 15824436 TI - The use of Xenopus oocytes and embryos as a route towards cell replacement. AB - When nuclei of somatic cells are transplanted to enucleated eggs of Xenopus, a complete reprogramming of nuclear function can take place. To identify mechanisms of nuclear reprogramming, somatic nuclei can be transplanted to growing meiotic oocytes of Xenopus, and stem cell genes activated without DNA replication. The combination of somatic cell nuclear transfer with morphogen signalling and the community effect may lead towards the possibility of cell replacement therapy. When mechanisms of nuclear reprogramming are understood, it may eventually be possible to directly reprogramme human somatic cell nuclei without the use of eggs. PMID- 15824437 TI - Collateral damage: spread of repeat-induced point mutation from a duplicated DNA sequence into an adjoining single-copy gene in Neurospora crassa. AB - Repeat-induced point mutation (RIP) is an unusual genome defense mechanism that was discovered in Neurospora crassa. RIP occurs during a sexual cross and induces numerous G : C to A : T mutations in duplicated DNA sequences and also methylates many of the remaining cytosine residues. We measured the susceptibility of the erg-3 gene, present in single copy, to the spread of RIP from duplications of adjoining sequences. Genomic segments of defined length (1, 1.5 or 2 kb) and located at defined distances (0, 0.5, 1 or 2 kb) upstream or downstream of the erg-3 open reading frame (ORF) were amplified by polymerase chain reaction (PCR), and the duplications were created by transformation of the amplified DNA. Crosses were made with the duplication strains and the frequency of erg-3 mutant progeny provided a measure of the spread of RIP from the duplicated segments into the erg 3 gene. Our results suggest that ordinarily RIP-spread does not occur. However, occasionally the mechanism that confines RIP to the duplicated segment seems to fail (frequency 0.1-0.8%) and then RIP can spread across as much as 1 kb of unduplicated DNA. Additionally, the bacterial hph gene appeared to be very susceptible to the spread of RIP-associated cytosine methylation. PMID- 15824438 TI - Stochasticity or the fatal 'imperfection' of cloning. AB - The concept of clone is analysed with the aim of exploring the limits to which a phenotype can be said to be determined geneticaly. First of all, mutations that result from the replication, topological manipulation or lesion of DNA introduce a source of heritable variation in an otherwise identical genetic background. But more important, stochastic effects in many biological processes may superimpose a phenotypic variation which is not encoded in the genome. The source of stochasticity ranges from the random selection of alleles or whole chromosomes to be expressed in small cell populations, to fluctuations in processes such as gene expression, due to limiting amounts of the players involved. The picture emerging is that the term clone is a statistical over-simplification representing a series of individuals having essentially the same genome but capable of exhibiting wide phenotypic variation. Finally, to what extent fluctuations in biological processes, usually thought of as noise, are in fact signal is also discussed. PMID- 15824439 TI - The return of the whole organism. AB - The long trend towards analysis at lower and lower levels is starting to reverse. The new integrative studies must make use of the resources uncovered by molecular biology but should also use the characteristics of whole organisms to measure the outcomes of developmental processes. Two examples are given of how movement between levels of analysis is being used with increasing power and promise. The first is the study of behavioural imprinting in birds where many of the molecular and neural mechanisms involved have been uncovered and are now being integrated to explain the behaviour of the whole animal. The second is the triggering during sensitive periods in early life by environmental events of one of several alternative modes of development leading to different phenotypes. A renewed focus on the whole organism is also starting to change the face of evolutionary biology. The decision-making and adaptability of the organism is recognized an important driver of evolution and is increasingly seen as an alternative to the gene-focused views. PMID- 15824440 TI - Do plants and animals differ in phenotypic plasticity? AB - This paper compares the flexibility in the nexus between phenotype and genotype in plants and animals. These taxa although considered to be fundamentally different are found to be surprisingly similar in the mechanisms used to achieve plasticity. Although non-cognitive behaviour occurs in plants, its range is limited, while morphological and developmental plasticity also occur to a considerable extent in animals. Yet both plants and animals are subject to unique constraints and thus need to find unique solutions to functional problems. A true comparison between the plant and animal phenotype would be a comparison between plants and sessile photosynthesizing colonial invertebrates. Such comparisons are lacking. However, they would provide important insights into the adaptive significance of plasticity in these groups. It is also suggested that a comparison of inflexible traits in these groups would provide an understanding of the constraints, as well as the costs and benefits,of a plastic versus non plastic phenotype in plants and animals. PMID- 15824441 TI - Not in their genes: phenotypic flexibility, behavioural traditions and cultural evolution in wild bonnet macaques. AB - Phenotypic flexibility, or the within-genotype, context-dependent, variation in behaviour expressed by single reproductively mature individuals during their lifetimes, often impart a selective advantage to organisms and profoundly influence their survival and reproduction. Another phenomenon apparently not under direct genetic control is behavioural inheritance whereby higher animals are able to acquire information from the behaviour of others by social learning, and, through their own modified behaviour, transmit such information between individuals and across generations. Behavioural information transfer of this nature thus represents another form of inheritance that operates in many animals in tandem with the more basic genetic system. This paper examines the impact that phenotypic flexibility, behavioural inheritance and socially transmitted cultural traditions may have in shaping the structure and dynamics of a primate society- that of the bonnet macaque (Macaca radiata), a primate species endemic to peninsular India. Three principal issues are considered: the role of phenotypic flexibility in shaping social behaviour, the occurrence of individual behavioural traits leading to the establishment of social traditions, and the appearance of cultural evolution amidst such social traditions. Although more prolonged observations are required, these initial findings suggest that phenotypic plasticity, behavioural inheritance and cultural traditions may be much more widespread among primates than have previously been assumed but may have escaped attention due to a preoccupation with genetic inheritance in zoological thinking. PMID- 15824442 TI - Mechanisms for the environmental regulation of gene expression: ecological aspects of animal development. AB - The environment can play a significant role in the production of phenotypes. However, the developmental mechanisms by which the environmental agents effect normal development are just becoming known. At least three paths have been found through which the environment can modify gene activity. The first is the neuroendocrine route. Here, the nervous system monitors the environment and transfers signals to the endocrine system. The endocrine hormones can then alter gene expression. The second route involves environmental factors that change the methylation pattern of genes, thereby altering their transcriptional capabilities. The third route involves the direct induction of gene expression in the host by microbial symbionts. The normal regulation of phenotype production by the environment should be considered a normal component of development and developmental biology. PMID- 15824443 TI - The pre-Mendelian, pre-Darwinian world: shifting relations between genetic and epigenetic mechanisms in early multicellular evolution. AB - The reliable dependence of many features of contemporary organisms on changes in gene content and activity is tied to the processes of Mendelian inheritance and Darwinian evolution. With regard to morphological characters, however, Mendelian inheritance is the exception rather than the rule, and neo-Darwinian mechanisms in any case do not account for the origination (as opposed to the inherited variation) of such characters. It is proposed, therefore, that multicellular organisms passed through a pre-Mendelian, pre-Darwinian phase, whereby cells, genes and gene products constituted complex systems with context-dependent, self organizing morphogenetic capabilities. An example is provided of a plausible 'core' mechanism for the development of the vertebrate limb that is both inherently pattern forming and morphogenetically plastic. It is suggested that most complex multicellular structures originated from such systems. The notion that genes are privileged determinants of biological characters can only be sustained by neglecting questions of evolutionary origination and the evolution of developmental mechanisms. PMID- 15824444 TI - Can morphogenesis be understood in terms of physical rules? AB - Because the morphogenesis of biological systems is not fully understood, researches from various points of view are necessary. The present author has recently made computer simulations with his colleagues to construct branching systems of human organs, such as the lung airway and the liver blood vessels. In the simulations certain rules are assumed to govern bifurcating processes of the systems. These rules are expressed in terms of physical and geometrical concepts, such as minimum energy consumption and uniform filling of branches in the space of organs. Results of computer simulation are quite similar to real structures. However, actual mechanisms of morphogenesis, i.e. effects of genes or proteins, are not considered in these studies. In this article, the present work is discussed in relation to the concept of biological pattern formation by Meinhardt and a recent study by Miura and Shiota on lung growth. PMID- 15824445 TI - Genotype, phenotype and cancer: role of low penetrance genes and environment in tumour susceptibility. AB - Role of heredity and lifestyle in sporadic cancers is well documented. Here we focus on the influence of low penetrance genes and habits, with emphasis on tobacco habit in causing head and neck cancers. Role of such gene-environment interaction can be well studied in individuals with multiple primary cancers. Thus such a biological model may elucidate that cancer causation is not solely due to genetic determinism but also significantly relies on lifestyle of the individual. PMID- 15824446 TI - Emergentism as a default: cancer as a problem of tissue organization. AB - During the last fifty years the dominant stance in experimental biology has been reductionism. For the most part, research programs were based on the notion that genes were in 'the driver's seat' controlling the developmental program and determining normalcy and disease (genetic reductionism and genetic determinism). Philosophers were the first to realize that the belief that the Mendelian genes were reduced to DNA molecules was questionable. Soon after these pronouncements, experimental data confirmed their misgivings. The optimism of molecular biologists, fueled by early success in tackling relatively simple problems, has now been tempered by the difficulties found when attempting to understand complex biological problems. Here, we analyse experimental data that illustrate the shortcomings of this sort of reductionism. We also examine the prevailing paradigm in cancer research, the somatic mutation theory (SMT), the premises of which are: (i) cancer is derived from a single somatic cell that has accumulated multiple DNA mutations; (ii) the default state of cell proliferation in metazoa is quiescence; and (iii) cancer is a disease of cell proliferation caused by mutations in genes that control proliferation and the cell cycle. We challenge the notion that cancer is a cellular problem caused by mutated genes by assessing data gathered both from within the reductionist paradigm and from an alternative view that regards carcinogenesis as a developmental process gone awry. This alternative view, explored under the name of the tissue organization field theory (TOFT), is based on premises that place cancer in a different hierarchical level of complexity from that proposed by the SMT, namely: (i) carcinogenesis represents a problem of tissue organization comparable to organogenesis, and (ii) proliferation is the default state of all cells. We propose that the organicist view, in which the TOFT is based, is a good starting point from which to explore emergent phenomena. However, new theoretical concepts are needed in order to grapple with the apparent circular causality of complex biological phenomena in development and carcinogenesis. PMID- 15824447 TI - Language from a biological perspective. AB - The faculty of language is unique to the human species. This implies that there are human-specific biological changes that lie at the basis of human language. However, it is not clear what the nature of such changes are, and how they could be shaped by evolution. In this paper, emphasis is laid on describing language in a Chomskyan manner, as a mental object. This serves as a standpoint to speculate about the biological basis of the emergence and evolution of language. PMID- 15824448 TI - Language cannot be reduced to biology: perspectives from neuro-developmental disorders affecting language learning. AB - The study of language knowledge guided by a purely biological perspective prioritizes the study of syntax. The essential process of syntax is recursion- the ability to generate an infinite array of expressions from a limited set of elements. Researchers working within the biological perspective argue that this ability is possible only because of an innately specified genetic makeup that is specific to human beings. Such a view of language knowledge may be fully justified in discussions on biolinguistics, and in evolutionary biology. However, it is grossly inadequate in understanding language-learning problems, particularly those experienced by children with neurodevelopmental disorders such as developmental dyslexia, Williams syndrome, specific language impairment and autism spectrum disorders. Specifically, syntax-centered definitions of language knowledge completely ignore certain crucial aspects of language learning and use, namely, that language is embedded in a social context; that the role of envrironmental triggering as a learning mechanism is grossly underestimated; that a considerable extent of visuo-spatial information accompanies speech in day-to day communication; that the developmental process itself lies at the heart of knowledge acquisition; and that there is a tremendous variation in the orthographic systems associated with different languages. All these (socio cultural) factors can influence the rate and quality of spoken and written language acquisition resulting in much variation in phenotypes associated with disorders known to have a genetic component. Delineation of such phenotypic variability requires inputs from varied disciplines such as neurobiology, neuropsychology, linguistics and communication disorders. In this paper, I discuss published research that questions cognitive modularity and emphasises the role of the environment for understanding linguistic capabilities of children with neuro-developmental disorders. The discussion pertains to two specific disorders, developmental dyslexia and Williams syndrome. PMID- 15824450 TI - Absence of strand bias for deletion mutagenesis during chromosomal leading and lagging strand replication in Escherichia coli. AB - Investigations were carried out to determine whether both DNA strands involved in Escherichia coli chromosomal DNA replication are replicated with similar accuracy. Experiments consisted of measuring the forward mutation rate from tonB(+) to tonB(-) in pairs of polA deficient strains in which the chromosomal target gene tonB was oriented in the two possible directions relative to the origin of replication, oriC. Within these pairs, the tonB sequence would be subjected to leading strand replication in one orientation and to lagging strand replication in the other. The most common tonB mutations in the polA1 strain were deletions followed by frameshifts. Among the deletions, a strong hotspot site with a 13-base deletion in the polA1 strains accounted for 18 of the 33 deletions in the one orientation, and 31 of the 58 deletions in the other. The results suggested that the two strands were replicated with equal or similar accuracy for deletion formation. PMID- 15824451 TI - Precise characterization of rDNA genes by intraspecies and inter-loci comparison of rDNA sequences and biochemical analysis of ribosomal RNA molecules in Agrobacterium tumefaciens. AB - Annotation of rRNA genes has been incomplete in Agrobacterium species although a number of Agrobacterial rDNA fragments have been sequenced. In this study, precise characterization of rRNA operons (rrn) was carried out in two biovar 1 strains, C58 and MAFF301001. Complete DNA sequencing of four rrns in MAFF301001 indicated that each operon codes for 16S, 23S and 5S rRNA as well as three tRNAs, trn(Ile), trn(Ala) and trn(Met). The genes and 16S-23S ITS of a given locus were exactly identical with those in the other three loci, except for a T-base loss in the 23S rRNA gene of rrnA and in the 5S rRNA gene of rrnB. Comparison with the four C58 rDNAs available in the DNA database indicated extensive sequence and size variations in the 23S rRNA gene, suggesting the presence of an intervening sequence (IVS). Biochemical RNA analysis, including Northern hybridization and 5' end mapping, in MAFF301001 revealed 2886-base and 2571-base precursors, two 1.3 kb major fragments, a 150-base fragment and removal of an IVS for 23S rRNA. We confirmed similar biochemical characteristics in the C58 strain. The features of rDNA detected here enable correction of previously reported information about Agrobacterial rRNAs and rRNA genes and should be useful for phylogenetic considerations. PMID- 15824452 TI - Improvement of the bioluminescence reporter system for real-time monitoring of circadian rhythms in the cyanobacterium Synechocystis sp. strain PCC 6803. AB - Circadian rhythm is a self-sustaining oscillation whose period length coincides with the 24-hour day-night cycle. A powerful tool for circadian clock research is the real-time automated bioluminescence monitoring system in which a promoter region of a clock-controlled gene is fused to a luciferase reporter gene and rhythmic regulation of the promoter activity is monitored as bioluminescence. In the present study, we greatly improved the bioluminescence reporter system in the cyanobacterium Synechocystis sp. strain PCC 6803. We fused an 805-bp promoter region of the dnaK gene seamlessly to the luxA coding sequence and integrated the P(dnaK)::luxAB fusion gene into a specific intergenic region of the Synechocystis genome (targeting site 1). The resulting new reporter strain, PdnaK::luxAB(-), showed 12 times the bioluminescence intensity of the standard reporter strain, CFC2. Furthermore, we generated strain PdnaK::luxAB(+), in which the P(dnaK)::luxAB fusion gene and the selection-marker spectinomycin resistance gene are transcribed in opposite directions. The PdnaK::luxAB(+) strain showed 19 times the bioluminescence intensity of strain CFC2. The procedures used to increase the bioluminescence intensity are especially useful for bioluminescence monitoring of genes with low promoter activity. In addition, these reporter constructs facilitate bioluminescence monitoring of any gene because the promoter fragments they contain can easily be replaced by digestion with unique restriction enzymes. They would therefore contribute to a genome-wide analysis of gene expression in Synechocystis. PMID- 15824453 TI - A catalytic subunit of cyclic AMP-dependent protein kinase, PKAC-1, regulates asexual differentiation in Neurospora crassa. AB - A cyclic AMP (cAMP)-dependent protein kinase pathway has been shown to regulate growth, morphogenesis and virulence in filamentous fungi. However, the precise mechanisms of regulation through the pathway remain poorly understood. In Neurospora crassa, the cr-1 adenylate cyclase mutant exhibits colonial growth with short aerial hyphae bearing conidia, and the mcb mutant, a mutant of the regulatory subunit of cAMP-dependent protein kinase (PKA), shows the loss of growth polarity at the restrictive temperature. In the present study, we isolated mutants of the catalytic subunit of the PKA gene pkac-1 through the process of repeat-induced point mutation (RIP). PKA activity of the mutants obtained through RIP was undetectable. The genome sequence predicts two distinct catalytic subunit genes of PKA, named pkac-1 (NCU06240.1, AAF75276) and pkac-2 (NCU00682.1), as is the case in most filamentous fungi. The results suggest that PKAC-1 works as the major PKA in N. crassa. The phenotype of the pkac-1 mutants included colonial growth, short aerial hyphae, premature conidiation on solid medium, inappropriate conidiation in submerged culture, and increased thermotolerance. This phenotype of pkac-1 mutants resembled to that of cr-1 mutants, except that the addition of cAMP did not rescue the abnormal morphology of pkac-1 mutants. The loss of growth polarity at the restrictive temperature in the mcb mutant was suppressed by pkac 1 mutation. These results suggest that the signal transduction pathway mediated by PKAC-1 plays an important role in regulation of aerial hyphae formation, conidiation, and hyphal growth with polarity. PMID- 15824454 TI - Fluorescent labeling of plant chromosomes in suspension by FISH. AB - By optimizing the concentration and time of treatment with hydroxyurea (HU), a DNA synthesis inhibitor, and trifluralin, a microtubule inhibitor, a highly effective (over 60%) cell cycle synchronization method for rye and barley meristem cells was developed. Chromosome suspensions containing highly purified and morphologically intact rye and barley chromosomes were prepared from the meristems of their root tips by homogenization. Digoxigenin-labeled 5S rDNA was used as a probe in FISH for the rye chromosomes in the suspension, and biotin labeled 17S rDNA and centromeric DNA were used in FISH for the rye and barley chromosome suspensions, respectively. Bright signals were detected at the specific regions of interest on the chromosomes. The results indicate that the method developed in this study is useful for selection and sorting of chromosomes that are not distinguishable by other means, using specific fluorescent labeling by FISH of the chromosomes in suspension. PMID- 15824455 TI - Alternative splicing of Pot1 (Protection of telomere)-like genes in Arabidopsis thaliana. AB - The Pot1 (Protection of telomere 1) is a G-rich single-stranded telomeric DNA binding protein, identified first in Schizosaccharomyces pombe, and shown to play an important role in stabilizing chromosomes. Pot1-like proteins or their encoding genes have been identified from yeasts to mammals. Based on the N terminal amino acid sequences of fission yeast and human Pot1, two Pot1-like proteins (AtPOT1-1 and AtPOT1-2) have been identified in Arabidopsis thaliana, but neither of them has been characterized yet. In this study, we amplified their full-length cDNAs by RT-PCR and found three different variants for AtPOT1-1 and two for AtPOT1-2 genes, suggesting that they are exposed to alternative splicing. Alternative splicing also occurs in human Pot1, and only one out of five splicing variants had tissue specificity. However, no tissue specificity was found for any variants of the AtPOT1-1 and AtPOT1-2 genes among buds, flowers, leaves, roots, stems, siliques and cultured cells. Northern blot hybridization indicated that AtPOT1-1 expresses more in meristematic tissues than in vegetative tissues. By western blot analysis, we found that the antibody made against the N-terminal amino acids of AtPOT1-1 recognized three different polypeptides, indicating that all three variants are being translated in Arabidopsis. PMID- 15824456 TI - Chromosome restriction enzyme digestion in domestic pig (Sus scrofa) constitutive heterochromatin arrangement. AB - The bimodal karyotype of pig appears to contain two types of constitutive heterochromatin, reflecting different satellite DNA families: GC-rich heterochromatin located mainly in the centromeric regions of the biarmed chromosomes, and less-GC-rich heterochromatin in the centromeric regions of the one-armed chromosomes. In order to better discriminate this constitutive heterochromatin, we treated pig chromosome preparations with eight different restriction endonucleases, followed by C-banding. This technique allowed an expedited characterization of the constitutive heterochromatin and demonstrated its great heterogeneity in pig chromosomes. Our work allowed the detection and identification of twenty-two heterochromatin subclasses (twelve centromeric, four interstitial, five telomeric, and the Yq band). Moreover, several cryptic interstitial and telomeric bands were revealed. The work presented here is useful not only for fundamental studies of chromosome banding and constitutive heterochromatin, but also offers a new approach for pig clinical cytogenetics. PMID- 15824457 TI - Microsatellite analysis of the spectacled bear (Tremarctos ornatus) across its range distribution. AB - DNA samples of the spectacled bear (Tremarctos ornatus) from five Andean countries, Venezuela, Colombia, Ecuador, Peru and Bolivia, were analyzed for nine microsatellite loci. Seven of them were polymorphic, which led us to investigate several population-genetic parameters. Private alleles and significant differences in gene frequencies were found among the populations studied, which demonstrated the extent of genetic differentiation among the spectacled bear populations. The levels of gene diversity measured with these microsatellites were rather modest in this species. Hardy-Weinberg disequilibrium was especially found for the overall and the Ecuadorian samples, and might be due to the Wahl und effect or consanguinity. Significant genetic heterogeneity was mainly observed among the Colombian and the Ecuadorian populations. Markov chain Monte Carlo simulations clearly showed that two different gene pools were present, one present in the Venezuelan-Colombian bears and other in the Ecuadorian ones. PMID- 15824458 TI - A comparison of interval mapping procedures including the QTL-cluster mapping. AB - Using the deterministic sampling, patterns of the log-likelihood surfaces expected in some interval mapping procedures for estimating the position of, and the effect for, QTL(s) were investigated for the situations where a single QTL or closely linked QTLs are contained in a chromosome segment bracketed with two markers. The mapping procedures compared were the conventional, likelihood-based interval mapping (IM), the regression interval mapping (RIM), and the QTL-cluster mapping (CM) in which the conditional probabilities of transmission of the whole segment marked by the flanking markers were taken into consideration. The half sib design was used here, and several cases of the true genetic model were considered, differing in the number of QTLs contained in the marker interval, the linkage phase for the sire, and the magnitude of the QTL(s) effect. For the true genetic models where a single QTL or closely linked QTLs being in coupling phase are contained in the interval, with (R)IM, clear global maxima of the log likelihood were observed within the range of the marker interval. It was shown that the estimates of the QTL(s) effect at the marked segment level are expected to be unbiased. On the other hand, in a setting where the linkage phase for the linked QTLs located in the interval was different from coupling and repulsion, there was found a ridge along the interval for the log-likelihood surface with (R)IM, indicating the dependency between the estimates of the position of, and the effect for, the putative QTL. For this case, it was found that the position of the putative QTL could be estimated as that of one of the flanking markers, and the estimate of the QTL effect be biased. In contrast, it was revealed that CM is expected to provide the unbiased estimate of the QTL(s)-effect at the segment level for any case of the true genetic models considered here. If the aim is for marker-assisted selection rather than mapping closely linked QTLs individually, then the CM approach is considered to be useful. PMID- 15824459 TI - Effective breakage of phage lambda DNA by shearing with ceramic-coated needle of syringe. AB - The loss of biological activity of phage lambda DNA was much greater when the DNA was sheared using a ceramic-coated needle attached to a syringe compared with a conventional stainless steel needle. Inactivation of the biological activity was due to breakage at the middle of the molecule. The thickness of the ceramic coating was a crucial factor for the breakage. Because approximately the same level of inactivation was observed with a non-coated needle as with thin glass and quartz tubes, it was concluded that the unknown characteristic(s) of the silicon nitride (SiNx) coating itself resulted in the effective breakage of lambda DNA molecules by shearing force. PMID- 15824460 TI - Pleiotropic actions of aldosterone and the effects of eplerenone, a selective mineralocorticoid receptor antagonist. AB - Aldosterone plays an important role in the pathogenesis of cardiovascular and renal disease that is independent of angiotensin II. Mineralocorticoid receptors are expressed in nonepithelial tissues such as the heart and blood vessels. Although mineralocorticoid receptor antagonism reduces mortality in patients with congestive heart failure, the progestational and antiandrogenic side effects of the nonspecific mineralocorticoid receptor antagonist, spironolactone, have limited its usefulness in the treatment of cardiovascular diseases. This review examines the expanding role of aldosterone, including its broad spectrum of non classical effects, and the recent clinical and experimental trials with the selective mineralocorticoid receptor antagonist, eplerenone. PMID- 15824461 TI - Role of interstitial ATP and adenosine in the regulation of renal hemodynamics and microvascular function. AB - The role of adenosine in the regulation of renal hemodynamics and function has been studied extensively; however, another purine agent, ATP, is also gaining recognition for its paracrine role in the kidney. Adenosine and ATP bind to specific membrane-bound P1 and P2 purinoceptors, respectively, and initiate a variety of biological effects on renal microvascular tone, mesangial cell function, and renal epithelial transport. The purpose of this review is to summarize the potential roles of interstitial ATP and adenosine as regulators of renal hemodynamics and microcirculation. In vitro blood-perfused juxtamedullary nephron preparation was used to assess the roles of ATP and adenosine in the regulation of renal microvascular tone. This approach mimics the adventitial exposure of renal microvascular smooth muscle to ATP and adenosine synthesized locally and released into the interstitial fluid. ATP selectively vasoconstricts afferent but not efferent arterioles via P2X and P2Y receptors, whereas, adenosine vasoconstricts both vascular segments via activation of adenosine A(1) receptors. Furthermore, selective P2X and P2Y receptor stimulation increases intracellular calcium concentration in vascular smooth muscle cells that are freshly isolated from the preglomerular microvasculature. These data support the hypothesis that interstitial ATP plays a critical role in the control of renal microvascular function through mechanisms that are independent of adenosine receptors. We have recently developed a renal microdialysis method to determine the dynamics of ATP and adenosine levels in the renal cortical interstitium. In this review, we also summarize current knowledge pertaining to the alterations in renal interstitial ATP and adenosine in some pathophysiological conditions. PMID- 15824462 TI - Predictors of controlled ambulatory blood pressure in treated hypertensive patients with uncontrolled office blood pressure. AB - Although some treated hypertensive patients have controlled 24-h ambulatory blood pressure (ABP) despite their uncontrolled office blood pressure (BP), the factors relating to the control of 24-h ABP remain unknown. We conducted a study to assess 24-h ABP and its association with other cardiovascular risk factors, including echocardiographic left ventricular hypertrophy (LVH), in elderly hypertensive patients (n =41) with uncontrolled office BP (>140/90 mmHg) during long-term medication. Although a majority of the patients had isolated elevation of office systolic BP (SBP), there was no significant relationship between office SBP and 24-h SBP, and about half of the patients had controlled 24-h ABP (125+/ 8/69+/-6 mmHg). Patients with controlled 24-h ABP (125+/-8/69+/-6 mmHg) had similar office BP (150+/-6/77+/-5 vs. 150+/-7/79+/-7 mmHg), but lower left ventricular mass index (LVMI) (123+/-34 vs. 156+/-34 g/m(2)) and body mass index (BMI) (24.4+/-2.1 vs. 26.4+/-3.6 kg/m(2)) compared with those with uncontrolled 24-h ABP (149+/-13/78+/-7 mmHg). Multivariate analysis showed that LVMI and BMI were independently associated with controlled 24-h ABP, and the control status of 24-h ABP was highly dependent on the presence of LVH and obesity. Therefore, absence of LVH and obesity may be useful for predicting the level of control of 24-h ABP in treated patients whose office BP is uncontrolled without ABP measurements. PMID- 15824463 TI - Peroxisome proliferator-activated receptor-gamma coactivator-1alpha polymorphism is not associated with essential hypertension and type 2 diabetes mellitus in Chinese population. AB - To investigate whether variations in the peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) are associated with essential hypertension and type 2 diabetes in a Chinese population. A case-control study design was applied in a Chinese population. Two single nucleotide polymorphisms (SNPs), +1302G>A and G482S, in the PGC-1alpha gene were genotyped and compared between 494 unrelated Chinese subjects with essential hypertension and type 2 diabetes and 555 normal control subjects with the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. These two polymorphisms were in highly significant linkage disequilibrium with each other (p <0.0001). The frequency of the 482S allele was 42.9% in the Chinese population, which was similar to the frequency in the Japanese population (43.7%), but much higher than those of Caucasian populations (30.8% to 38.1%). There were no associations of the G482S and +1302G>A polymorphisms and haplotype combinations with essential hypertension and type 2 diabetes. In addition, no associations were found between these two polymorphisms and blood pressure. In conclusion, these results indicated that these two variations in the PGC-1alpha gene might not contribute to the risk of hypertension and type 2 diabetes in the Chinese population studied here. PMID- 15824464 TI - The thiazide-sensitive Na(+)-Cl(-) cotransporter gene, C1784T, and adrenergic receptor-beta3 gene, T727C, may be gene polymorphisms susceptible to the antihypertensive effect of thiazide diuretics. AB - The response of blood pressure to thiazide diuretics (TZDs) differs among individuals. The prediction of the antihypertensive effect of TZDs is important for realizing individualized therapy in the management of hypertension. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) susceptible to the antihypertensive effect of TZDs, particularly focusing on genes related to water-electrolyte absorption in the kidney. Seventy-six outpatients (mean age, 65.4+/-9.0 years) with essential hypertension (EHT) taking TZDs were retrospectively assessed. We defined as responders (R) those whose mean blood pressure was lowered by more than 5 mmHg after the use of TZDs. Forty-eight SNPs in 17 genes (ADD1, GNB3, TSC [SLC12A3], MLR [NR3C2], NCX1 [SLC8A1], WNK1, WNK4, AGT, ACE, AT1 [AGTR1], CYP11B2, ADRB1, ADRB2, ADRB3, ADRA1A, ADRA1B, ADRA2A) were genotyped in the 76 patients. The SNPs in TSC, MLR, NCX1, WNK1, and WNK4 were identified by direct sequencing and those with minor frequencies of greater than 5% were genotyped in this study. The comparison of polymorphism prevalence between R and non-responders (NR) showed significant differences in TSC C1784T (C allele vs. T allele, odds ratio (OR)=3.81, p =0.016, confidence interval (CI): 1.25-11.63) and ADRB3 T727C (Trp64Arg) (T allele vs. C allele, OR=4.59, p =0.005, CI: 1.54-13.68). The blood pressure (BP) in patients homozygous for the major alleles of both TSC C1784T and ADRB3 T727C were significantly reduced by TZD treatment; however, the BP in those homozygous for the minor allele and heterozygous (TSC C1784T: TT+CT; ADRB3 T727C: CC+CT) for both SNPs were not significantly changed after TZD treatment. Both newly detected TSC C1784T and ADRB3 T727C are gene polymorphisms susceptible to the antihypertensive effect of TZDs in patients with EHT. Thus, the prediction of BP reduction by TZDs may be possible by evaluating these two SNPs. PMID- 15824465 TI - Hyperuricemia as a predictor of hypertension in a screened cohort in Okinawa, Japan. AB - Several epidemiological studies have shown a positive association between serum uric acid levels and the risk of hypertension. However, subjects in these studies were mostly men, or were incompletely examined for lifestyle-related variables. We prospectively examined the relation between hyperuricemia and the risk of developing hypertension with consideration for alcohol consumption and smoking habits in a large screened cohort of men and women. A total of 4,489 individuals (2,927 men and 1,562 women) who did not have hypertension and were not currently using antihypertensive medication were examined at the Okinawa General Health Maintenance Association in 1977. Subjects were re-examined in 2000. Hyperuricemia was defined as a serum uric acid level >or=7.0 mg/dl in men and >or=6.0 mg/dl in women. Hypertension was defined as systolic blood pressure (SBP) >or=140 mmHg, and/or diastolic blood pressure (DBP) >or=90 mmHg. A total of 289 subjects (201 men and 88 women) were hypertensive (SBP >or=140 mmHg, and/or DBP >or=90 mmHg) in 2000. Multivariate analysis was performed for development of hypertension in hyperuricemic subjects, adjusted for age, family history of hypertension, alcohol consumption, cigarette smoking, obesity, hypercholesterolemia, hypertriglyceridemia, low high-density lipoprotein cholesterol, and diabetes mellitus. The adjusted odds ratio (95% confidence interval) in men was 1.48 (1.08 2.02) and in women was 1.90 (1.03-3.51) (p <0.05, respectively). The results showed hyperuricemia to be a new predictor of hypertension development in both men and women. PMID- 15824466 TI - Applicability of the stages of change model for analyzing fruit and vegetable intake in relation to urinary potassium excretion: baseline results from the High Risk and Population Strategy for Occupational Health Promotion (HIPOP-OHP) Study. AB - The Stages of Change model evaluates and conceptualizes attempts to alter particular behavior patterns. To investigate the validity of this model for assessing fruit and vegetable intake, we examined the association between the stage of change in fruit and vegetable intake and urinary potassium excretion. The data were from baseline surveys taken in 1999 and 2000 from the High-Risk and Population Strategy for Occupational Health Promotion (HIPOP-OHP) study in Japan. This was a non-randomized control trial at 12 worksites in Japan and aimed to decrease cardiovascular risk factors. Cross-sectional analysis was performed using data from 6,774 participants (5,364 men and 1,410 women). We used three categories of the model: precontemplation or contemplation (P/C), indicating no commitment to change; preparation (P), indicating readiness to change behavior but not actually doing so; and action or maintenance (A/M), indicating an actual change in behavior. Urinary potassium excretion was estimated from the potassium and creatinine concentrations in spot urine samples. Multivariate analysis indicated that urinary potassium excretion in the A/M stage was 1.65 mmol/day more than in the P stage, and 1.44 mmol/day more than in the P/C stage for men (p <0.05, respectively). For women, urinary potassium excretion in the A/M stage was 1.26 mmol/day more than in the P/C stage (p <0.05) and 1.04 mmol/day more than in the P stage, although the latter result lacked statistical significance (p =0.08). This study supports the potential value of the Stages of Change model for increasing fruit and vegetable intake in the design of dietary intervention programs. PMID- 15824467 TI - Reproducibility of arterial stiffness indices (pulse wave velocity and augmentation index) simultaneously assessed by automated pulse wave analysis and their associated risk factors in essential hypertensive patients. AB - Arterial stiffness is a strong determinant of cardiovascular risk. Pulse wave velocity (PWV) and the augmentation index (AIx) are widely used as arterial stiffness indices. We studied the reproducibility of these indices and their association with cardiovascular risk factors in hypertensives. We measured brachial blood pressure (BP), brachial-ankle PWV (baPWV) and carotid AIx (cAIx) twice (at the baseline and 4 weeks after the baseline) using an automatic device in 103 hypertensives. The mean intraobserver-intersession difference was 29.0 cm/s with an SD of 201.6 cm/s for baPWV, and 0.5% with an SD of 5.9% for cAIx, and the Bland-Altman plots demonstrated the good reproducibility of baPWV and cAIx. Both baPWV and cAIx (the average of the 1st and the 2nd measurements) were significantly correlated with age, systolic BP (SBP), and pulse pressure (all, p <0.005); however, these factors were not correlated with each other (r =0.06, NS). cAIx was correlated with height, heart rate (HR), total cholesterol, and low density lipoprotein cholesterol (LDL-C) (all, p <0.05). In multiple regression analysis, age, SBP, and HR emerged as significant independent predictors of baPWV (adjusted R(2)=0.43, p <0.0001), while height, SBP, HR, and LDL-C emerged as significant independent predictors of cAIx (adjusted R(2)=0.58, p <0.0001). Both PWV and AIx measured using an automatic device were fairly reproducible, and their associated risk factors appeared to be different. Automated simultaneous measurement of these arterial stiffness indices may be useful for risk stratification of hypertensives. PMID- 15824468 TI - Efficacy of a multicomponent program (patient-centered assessment and counseling for exercise plus nutrition [PACE+ Japan]) for lifestyle modification in patients with essential hypertension. AB - With conventional lifestyle modification programs, it can be difficult for hypertensive individuals to modify their lifestyles and maintain the changes. We assessed whether a multicomponent program (patient-centered assessment and counseling for exercise plus nutrition [PACE+ Japan]) based on behavior theory and social cognitive theory would be effective for treating patients with essential hypertension. We examined 57 outpatients aged 62+/-10 years with essential hypertension irrespective of antihypertensive drug treatment. Participants were randomly divided into two groups: 1) a PACE+ Japan follow-up group (n =18), who were given an action-plan sheet and systemic health counseling by a physician and counselor every 4 weeks for 24 weeks, and 2) a PACE+ Japan only group (n =20), who were given an action-plan sheet but did not receive counseling. An age- and sex-matched control group (n =19) was also selected. The decrease in systolic blood pressure (SBP) (Delta SBP=SBP at 24 weeks minus that at 0 weeks) in the PACE+ Japan follow-up group was significantly greater than that in the control group. In addition, the Delta percentage of Fat (%Fat) and Delta urinary sodium extraction (U-Na) in the PACE+ Japan follow-up group were significantly greater than those in the control group. With regard to changes in total energy expenditure, exercise energy expenditure and total energy intake between 0 weeks and 24 weeks, significant improvements were observed for the PACE+ Japan follow-up group. Delta U-Na was determined to significantly predict Delta SBP as assessed by stepwise selection. In addition, the partial correlation coefficient of Delta SBP with Delta U-Na was 0.361 (p =0.011) as assessed by a multiple regression analysis. Therefore, PACE+ Japan follow-up counseling was associated with a reduction in SBP, which in turn was associated with reduction in U-Na. This new program may be effective for reducing blood pressure in hypertensives. PMID- 15824469 TI - Effects of bradykinin on cardiovascular remodeling in renovascular hypertensive rats. AB - Angiotensin converting enzyme (ACE) inhibitors inhibit both the formation of angiotensin II and the catabolism of bradykinin (BK). They prevent not only hypertension but also cardiac hypertrophy and fibrosis. An increase in BK level stimulates the expression of nitric oxide (NO) synthase (NOS) and induces prostaglandins, both of which are powerful vasodilator factors. The direct effect of BK against cardiac hypertrophy is still unclear. This study was performed to examine the cardioprotective effects of BK in hypertrophic models. Renovascular hypertensive (RHT) rats were treated with BK (1,000 ng/kg/day), BK+D-arginyl [Hyp(3), Thi(5), D-Tic(7), Oic(8)]-bradykinin (HOE140) (a BK B(2) receptor antagonist), and BK+N(omega)-nitro-L-arginine methyl ester (L-NAME) (a NOS inhibitor) for 3 weeks. Blood pressure was measured and echocardiographic analysis performed during the treatment. Histological data were analyzed to confirm the hypotrophic effect of BK. Treatment with BK improved cardiac remodeling, reducing both the heart weight/body weight ratio and the left ventricular wall thickness. However, co-treatment with HOE140 or L-NAME reversed the anti-hypertrophic action of BK. In particular, cardiac fibrosis or perivascular fibrosis, along with collagen accumulation, were inhibited by treatment with BK, while HOE140 and L-NAME counteracted these changes. In addition, expressions of atrial natriuretic peptides (ANP) and brain natriuretic peptides (BNP), which are markers of cardiac abnormalities, were down-regulated by treatment with BK. These effects were reversed by co-treatment with HOE140 and L-NAME. Together, these results indicate that BK directly inhibits the progression of cardiac hypertrophy and cardiac fibrosis due to NO release via the BK B(2) receptor. The BK-NO pathway may play an important role in the progression of cardiac remodeling. PMID- 15824470 TI - Calcium antagonist reduces oxidative stress by upregulating Cu/Zn superoxide dismutase in stroke-prone spontaneously hypertensive rats. AB - Recent studies have suggested that the calcium antagonists have an antiatherogenic antioxidant property. The effects of the calcium antagonists on reactive oxygen species (ROS)-related enzymes, however, remain unknown. We hypothesized that the calcium antagonists inhibit oxidative stress in the hearts of stroke-prone spontaneously hypertensive rats (SHRSP) through the ROS scavenging enzymes known as superoxide dismutases (SODs). Male 12-week-old Wister Kyoto rats (WKY) and SHRSP were used for the study. SHRSP were randomized and treated for 6 weeks with a vehicle, amlodipine (5 mg/kg/day), or enalapril (10 mg/kg/day). NAD(P)H oxidase activity was measured by a luminescence assay, and SOD activity was measured spectrophotometrically. Protein expressions were analyzed by immunoblots. Both drugs showed equipotent effects on systolic blood pressure, left ventricular hypertrophy and fibrosis, the wall-to-lumen ratio, the manganese SOD activity, ROS, and the endothelial NO synthase expression in the SHRSP hearts. Furthermore, amlodipine significantly restored copper/zinc containing SOD (Cu/ZnSOD) expression and its activity in SHRSP hearts to a level equal to that of WKY more effectively than did enalapril (p <0.05), whereas enalapril downregulated NAD(P)H oxidase activity more than did amlodipine (p <0.05) in the SHRSP hearts. Furthermore, amlodipine restored Cu/ZnSOD expression and its activity in SHRSP hearts to a level equal to that in WKY hearts, and this restoration was significantly more effective than that by enalapril (p <0.05); on the other hand, enalapril induced a greater downregulation of NAD(P)H oxidase activity in SHRSP hearts than did amlodipine (p <0.05). Thus, amlodipine may inhibit vascular remodeling and oxidative stress in the SHRSP heart by efficiently upregulating Cu/ZnSOD, suggesting that the calcium antagonist may exhibit an antiatherogenic antioxidative action beyond blood-pressure lowering through the restoration of Cu/ZnSOD activity in the heart in cases of hypertension. PMID- 15824471 TI - Hepatocyte growth factor stimulates nitric oxide production through endothelial nitric oxide synthase activation by the phosphoinositide 3-kinase/Akt pathway and possibly by mitogen-activated protein kinase kinase in vascular endothelial cells. AB - Hepatocyte growth factor (HGF) has recently been the focus of attention due to its angiogenic effects, which are similar to those of vascular endothelial growth factor (VEGF); because of these effects, HGF is considered to be a novel therapeutic agent against vascular disorders, including atherosclerotic angiopathies. Although nitric oxide (NO), which is derived from vascular endothelial cells (ECs), is also involved in angiogenesis, little is known regarding the interactions between HGF and NO. We therefore examined the effects of HGF on NO production as well as endothelial NO synthase (eNOS) phosphorylation, and investigated their mechanisms. In bovine aortic ECs, HGF induced a rapid (5 min) increase of NO production measured by diaminofluorescein 2 diacetate. Moreover, HGF rapidly (2.5 min) stimulated eNOS phosphorylation (Ser 1179) as determined by Western immunoblot analyses. Both of these effects were almost completely suppressed by the phosphoinositide 3-kinase (PI3K) inhibitor LY294002, and were partially suppressed by the mitogen-activated protein kinase (MAPK) kinase 1/2 inhibitor U0126. HGF also stimulated Akt phosphorylation (Ser 473), which was completely suppressed by LY294002 and was partially suppressed by U0126. Moreover, HGF stimulated extracellular signal-regulated kinase 1/2 phosphorylation (Thr-202/Tyr-204), which was completely suppressed by U0126 and was partially suppressed by LY294002. Taken together, these results indicate that HGF not only phosphorylates eNOS through the PI3K/Akt pathway, but also partially through the MAPK pathway, and that these two pathways may interact. Compared with VEGF, HGF was more potent in both NO production and eNOS phosphorylation. Our study thus demonstrates a novel activity of HGF-the stimulation of NO production which occurs via eNOS phosphorylation that may in turn be mediated by cross-talk between the PI3K/Akt and MAPK pathways. PMID- 15824472 TI - Shiftwork and metabolic risk factors of cardiovascular disease. AB - We conducted this study to examine the relationship between shift work duration and the metabolic risk factors of cardiovascular disease among shift workers. The study population consisted of 226 female hospital nurses and 134 male workers at a firm manufacturing diapers and feminine hygiene materials, whose mean ages were 28.5 yr for the nurses and 29.1 yr for the male workers. The fasting blood sugar level, serum cholesterol, blood pressure, height and weight, waist and hip circumferences (only for the nurses), and numbers of walks during work (as a measure of physical activity) were measured. Using the Karasek's job contents questionnaire, job stress was assessed. Information about the years of work, shift work duration, past medical and behavioral history, including smoking, was obtained by a self-administered questionnaire. With definitions of hypertension as systolic blood pressure (SBP) > or =160 or diastolic blood pressure (DBP) > or =90 mmHg occurring at least once, hypercholesterolemia as serum total cholesterol > or =240 mg/dl, obesity as body mass index (BMI) > or =25 kg/m(2) and as waist to hip ratio (WHR) > or =0.85, we examined the prevalences of metabolic risk factors among subjects. Regression analyses to show the relationships between shift work duration and metabolic risk factors were performed using simple and multivariate models stratified by age, and adjusted for smoking, drinking, job strain and physical activity. Duration of shift work was significantly associated with SBP or cholesterol level among male workers aged 30 or more. Among female nurses, it was inversely associated with DBP (in those who were below 30 yr old) and cholesterol (in those who were aged 30 or more). BMI was non-significantly associated with the duration of shift work in both male workers and female nurses who were 30 yr old or more. WHR in female nurses increased slightly according to increasing duration of shift work. Fasting blood sugar was not significantly associated with the duration of shift work in either sex regardless of age-group. These results suggest an association between shift work duration and the metabolic risk factors of cardiovascular disease. PMID- 15824473 TI - Memantine alleviates toxicity induced by dichlorvos in rats. AB - The changes of N-methyl-D-aspartate (NMDA) receptor and protective efficacy of memantine (MEM) in rats poisoned with dichlorvos were studied. Dichlorvos evoked down-regulation of the affinity and density of [(3)H]MK-801 binding to NMDA receptor in the brain of rats receiving dichlorvos (15 and 25 mg/kg bw, i.p.). The binding capacity of NMDA receptor and acetylcholinesterase activity were determined at 4 h, 8 h, 16 h, 24 h and 48 h after treatment. When rats were given a different doses of MEM (5, 15 and 45 mg/kg bw) after poisoning (dichlorvos 25 mg/kg bw), the latency of onset of signs was postponed and the magnitude of muscular fasciculation was alleviated as the dose of MEM increased. The lower doses of MEM (5 and 15 mg/kg bw) could antagonize the dichlorvos-evoked down regulation of NMDA receptor, while the highest dose (45 mg/kg bw) decreased the Bmax and Kd values of NMDA receptors. These results show the dichlorvos-evoked down-regulation of NMDA receptor might be self-regulation by the body to protect the central nervous system. MEM could antagonize the down-regulation of NMDA receptors, and alleviated signs of poisoning, especially reducing the magnitude of muscular fasciculation. We suggest that the role of NMDA receptor in organophosphates (OP) poisoning should receive more attention, and, that MEM treatment in acute OP poisoning, as a supplement to atropine and oxime, should be considered. PMID- 15824474 TI - Job strain and risk of cardiovascular events in treated hypertensive Japanese workers: hypertension follow-up group study. AB - In Japan, Karoshi, which means "death from overwork", has become a focus of social concern. However, no previous study has examined long working hours and job strain simultaneously in relation to incidence of cardiovascular events (CVE) in Japanese workers. We prospectively evaluated the relation between job strain or long working hours and risk of CVE in treated hypertensives during the years 1994-2000. In this study, we followed a sample of 1,615 participants (908 men and 707 women) aged 40-65 yr who were working more than 5 working hours per day. Participants completed questions about work-related conditions and lifestyle variables at baseline, and were free of diagnosed cardiovascular disease, stroke or cancer. Job strain (the combination of job demands and job control) was assessed using a simple questionnaire developed with reference to Karasek's model. The main outcome measure was incidence of CVE. During the mean follow-up period of 5.6 yr, we documented 38 cases of initial CVE. Analyses were conducted using the Cox proportional hazard model. We found a significant association between incidence of CVE and job strain categories, but not long working hours. Multivariate relative risks for CVE in active jobs and high strain jobs overall were 2.89 (95% CI: 1.33-6.28) and 2.45 (95% CI: 0.87-6.93); for men 2.94 (95% CI: 1.29-6.73) and 1.86 (95% CI: 0.51-6.75), and for women 3.97 (95% CI: 0.34-46.88) and 9.05 (95% CI: 1.17-69.86), respectively. In conclusion, active jobs and high strain jobs were associated with increased risk of CVE for treated hypertensive workers. PMID- 15824475 TI - Work, unemployment and life satisfaction among patients with diisocyanate induced asthma--a prospective study. AB - Occupational asthma has been found to be associated with lowered socio-economic outcomes, an increased unemployment rate and a decreased quality of life. The compensation of occupational diseases is comparatively favourable in Finland. Our aim was to follow-up the working status and life satisfaction of patients with diisocyanate-induced asthma in 245 cases diagnosed during 1976-1992. A questionnaire was sent out on average 10 (3-19) yr after the diagnosis to the surviving 235 patients. The questionnaire was validated by re-examining 91 of them clinically, and with spirometry, histamine challenge test and peak flow surveillance. Of the 213 responding patients, 14% were unemployed, and for 50% of them unemployment was caused by asthma. Unemployment was associated with nocturnal asthma symptoms (OR 10.93; CI 2.69-44.452), increased PEF variability (OR 8.46; CI 1.52-46.97) and with the use of short-acting beta-sympathomimetic medication (p=0.045). Satisfaction with life was associated with present working (OR 3.50; CI 1.73-7.06) and with good condition of asthma as assessed by the use of asthma medication (OR 0.49; CI 0.27-0.89) and objective measurements of the asthma condition, e.g. PEF variability (OR 0.21; CI 0.08-0.59). As a conclusion, unemployment was remarkably low as compared with earlier reports and related to the period of simultaneous economic recession. Unemployment, as well as dissatisfaction with life were associated with poor conditions of asthma. Unemployment was associated with improper asthma care favouring the use of short acting beta-sympathomimetic medication. Proper follow-up of asthma is essential for minimizing the social complaints of occupational asthma, as well as for enhancing life satisfaction. PMID- 15824476 TI - The relationship of job type to burnout in social workers at social welfare offices. AB - This study sought to determine the relationship of job type differences to burnout level, the details of job characteristics for each job type, and the association between burnout and job factors in 189 social workers at all social welfare offices in a prefecture in Japan. Among the three job types, 32.9% of social workers involved with public assistance, 29.0% of social workers involved with public assistance, the elderly, the disabled and single mothers, and 15.2% of social workers involved with the elderly, the disabled and single mothers were scored in the "high burnout" category (p<.05). Job type differed significantly with respect to the job characteristics of percentage of time spent on home visits per typical working day, job satisfaction, aversion to the job, and social support. The job type of public assistance work had a higher percentage of time spent on home visits per typical working day, aversion to the job, lower job satisfaction, and less social support than the job type involving no public assistance work. Multiple regression analyses showed the associations between job factors and burnout for each job type. Aversion to the job had a primary positive association with burnout for all social worker job types. Social support had a negative association with burnout in social workers whose clients included public assistance cases. The number of years in social work had a negative association with burnout, while percentage of time spent doing interviews per typical working day had a positive association with burnout in social workers who were involved with public assistance, the elderly, the disabled and single mothers. These results suggest that the job type of public assistance work may carry a higher risk of burnout than job types involving no public assistance work. To ameliorate this risk, it was thought to be important to improve aversion to the job as well as having a social support network for public assistance social workers. PMID- 15824477 TI - Prevalence of the metabolic syndrome using the modified ATP III definitions for workers in Japan, Korea and Mongolia. AB - A clustering of insulin resistance, hypertension and dyslipidemia has been labeled as the metabolic syndrome. Asians have a lower frequency of obesity than do Caucasians, but have an increasing tendency toward metabolic syndrome. Most data on metabolic syndrome are based on studies from Western countries with only limited information derived from Asian populations. We conducted a cross sectional study of individuals aged 30-60 yr in workplace settings. We examined and analyzed the health data of 1,384 Japanese, Koreans and Mongolians for metabolic syndrome based on the modified definitions of the working definition proposed by the Third Report of the National Cholesterol Educational Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (ATP III definition). The prevalence of metabolic syndrome using the ATP III-BMI30 and ATP III-BMI25 definitions was 7% and 12% for Japanese, 7% and 13% for Koreans, and 12% and 16% for Mongolians, respectively. With the exception of obesity, the prevalences of individual metabolic abnormalities within each of the three Asian groups were similar to each other and to reported rates of prevalence in the U.S.A. Nevertheless, the values of sensitivity and specificity by the metabolic syndrome definitions are remarkably different relative to ethnicity. A universal metabolic syndrome definition is inappropriate for comparisons of metabolic syndrome among Asian ethnic groups. We believe that the ATP III-BMI25 definition is suitable for the determination of metabolic syndrome among Japanese and Koreans, and that the ATP III-BMI30 is more appropriate for Mongolians. PMID- 15824478 TI - The Check Bus Project and its effectiveness on health promotion at work. AB - Employees of two large companies in Switzerland took part in a nationwide health information and promotion campaign at work, which included various physical examinations. A total of 5,849 consecutive participants were sent a questionnaire to determine whether changes had been made in their lifestyles and if they had benefited from participation; 2,361 (40.4%) responded. The campaign was rated highly at 8.3 on a scale from 1 to 10. Respondents reported benefiting most from the personal examination results, in particular the measurement of bone density, examination of the carotid arteries and discussions with the staff. Of the respondents, 921 stated they changed their lifestyle in the area of physical activity (64.4%) and nutrition (63.6%). Multiple logistic regression analysis showed the variables predictive of responding were age, hierarchy, diet and smoking. Health promotion at work and the choice of offered examinations may influence the effects of health care promotion at the workplace and the readiness of participants to improve their attitudes to health. Health promotion at work is highly regarded by employees who pay more attention to diet, smoke less and in particular those over 50 yr of age. High-tech equipment in a mobile unit may attract employers and employees may profit most from the individual examination results; 39% of the respondents stated they changed their lifestyle as a consequence of the Check Bus campaign. PMID- 15824479 TI - Effects of mixed organic solvents on neuromotor functions among workers in Buddhist altar manufacturing factories. AB - To clarify the neuromotor effects of long-term exposure to mixed organic solvents, postural sway and tremor were measured in 62 solvent workers of four Buddhist altar manufacturing factories who had worked for 1-46 (mean 12) yr. By using the passive gas sampler, 8-h time-weighted average concentrations in the workers were estimated to be 0.02-8.7 ppm for toluene, 0.02-7.7 ppm for xylene, 0.02-5.5 ppm for styrene and 0.02-40.5 ppm for n-hexane. Sagittal sway and sway area of the posturography with eyes closed were significantly larger in the solvent workers than in 35 age-matched controls (p<0.05), and there was a significant difference in Romberg quotient of sagittal sway between the two groups (p<0.05). Also, tremor intensities at 1.0-5.9 Hz, 6.0-9.9 Hz and 10.0-13.9 Hz with the right hand, and at 6.0-9.9 Hz with the left hand were significantly stronger in the solvent workers than in the controls. Among the solvent workers, transversal and sagittal sways with eyes open and tremor intensity at 10.0-13.9 Hz were significantly related to toluene exposure (p<0.05), which may have been due to the acute effects of such solvents. These findings suggest that long-term exposure to mixed organic solvents may impair neuromotor functions as measured by postural sway and tremor. PMID- 15824480 TI - The associations between menstrual function and life style/working conditions among nurses in Taiwan. AB - This study was aimed at understanding the relationship among menstrual pattern, dysmenorrhea, life style and working conditions in nurses. The nurses were randomly selected from a medical center in Northern Taiwan. Each subject completed daily records including life and working conditions during the study period. The study showed that there were statistically significant differences in work years, daily working hours and type of work shift among nurses that worked at different units in the hospital. In the perceived regular cycle group, nurses that worked the night shift only exhibited the shortest menstrual cycles, less than 25 d. There was a significant difference (p<0.05) among the nurses' menstrual cycles. Many life factors (such as passive tobacco smoke exposure, perceived life satisfaction and perceived life stress) and working factors (such as work years, perceived work satisfaction and perceived work stress) were not significantly related to menstrual cycle regularity. In addition, 30% of the nurses complained of dysmenorrhea. Some factors including age, marital status and perceived life satisfaction were significantly related to dysmenorrhea. However, other life factors (such as passive tobacco smoke exposure, smoking, coffee, alcohol, cold drink habits, exercise and perceived life stress) and working factors (such as working places, type of work shift, daily work hours, perceived work satisfaction and perceived work stress) showed no correlation with dysmenorrhea. This study indicates that women should pay attention to their menstrual function and dysmenorrhea phenomenon. PMID- 15824481 TI - Karojisatsu in Japan: characteristics of 22 cases of work-related suicide. AB - With the rapidly increasing number of work-related suicides in Japan (Karojisatsu, in Japanese), both applications for worker's compensation insurance and civil suits are proliferating. The phenomenon of work-related suicide is examined along with the process and related factors. With informed consent from bereaved families, two certified psychiatrists independently reviewed and summarized 22 insurance and legal reports filed by psychiatrists on employee suicides that were related to heavy workloads. A clinical epidemiologist participated in discussions with psychiatrists to reach a consensus concerning the cause of the suicides. Only one case involved a female. Seventeen had experienced personnel changes, such as a promotion or transfer. Low social support was recognized in 18, high psychological demand in 18, low decision latitude in 17, and long working hours in 19 cases (more than 11 hours per day for 3 months or more, and without a day off in 9). The subjects had depressive episodes by the ICD-10 criteria and showed suicidal signs. Ten of them saw a general practitioner because of unspecified somatic complaints, but no effective measures were taken. None of them had a history of psychiatric consultation or had received mental health education dealing with job stress management. Although causality cannot be made from this case series report, we hypothesize that long working hours, heavy workloads, and low social support may cause depression, which can lead to suicide. Appropriate countermeasures are urgently needed and the present findings suggest some of them are possible. PMID- 15824482 TI - Hand arm vibration syndrome among quarry workers in Vietnam. AB - Few studies have focused on the health effects of vibrating tools on workers in the tropical area. Work conditions and health effects related to rock drill operation were studied in 102 quarry workers, including 73 rock drill operators in Vietnam. We aimed to clarify (1) risk of vibration exposure, (2) occurrence of vibration-induced white finger (VWF), and (3) characteristics of hand-arm vibration syndrome (HAVS). Total weighted r.m.s. acceleration of the Chinese -or Russian-made rock drills, was 45-55 m/s(2). According to work observation studies, daily exposure time to vibration was 160-210 min. ISO5349 predicted that this exposure level would be associated with a high risk of HAVS in workers. We found no clear evidence of VWF. There may be several reasons why no worker exhibited VWF: (1) warmer work conditions, (2) younger and less experienced workers, (3) seasonal changes in work operations, and (4) healthy worker effect. On the other hand, 5-10% of rock drill operators might be suffering from moderate HAVS which was sensori-neural type dominant. There may be some characteristic features of HAVS among quarry workers in the tropical area. PMID- 15824483 TI - Cancer incidence among workers potentially exposed to chlorinated solvents in an electronics factory. AB - A retrospective cohort morbidity study based on standardized incidence ratios (SIRs) was conducted to investigate the possible association between exposure to chlorinated organic solvents and various types of cancers in an electronics factory. The cohort of the exposed group was retrieved from the Bureau of Labor Insurance (BLI) computer database records dating from 1978 through December 31, 1997. Person-year accumulation began on the date of entry to the cohort, or January 1, 1979 (whichever came later), and ended on the closing date of the study (December 31, 1997), if alive without contracting any type of cancers, or the date of death, or the date of the cancer diagnosis. Vital status and cases of cancer of study subjects were determined from January 1, 1979 to December 31, 1997 by linking cohort data with the National Cancer Registry Database. The cancer incidence of the general population was used for comparison. After adjustment for age and calendar year, only SIR for breast cancer in the exposed female employees were significantly elevated when compared with the Taiwanese general population, based on the entire cohort without exclusion. The SIR of female breast cancer also showed a significant trend of period effect, but no significant dose-response relationship on duration of employment. Although the total cancer as well as the cancer for trachea, bronchus and lung for the entire female cohort was not significantly elevated, trend analysis by calendar-year interval suggested an upward trend. However, when duration of employment or latency was taken into consideration, no significantly elevated SIR was found for any type of cancer in either male or female exposed workers. In particular, the risk of female breast cancer was not indicated to be increased. No significant dose-response relationship on duration of employment and secular trend was found for the above-mentioned cancers. This study provides no evidence that exposure to chlorinated organic solvents at the electronics factory was associated with elevated human cancers. Dominant short-term employees may bias the cancer risk toward false positive. PMID- 15824484 TI - Environmental mycological study and allergic respiratory disease among tobacco processing workers. AB - This study presents the results of an investigation of respiratory symptoms, lung function, chest X-ray examinations, analysis of specific IgE antibodies and skin prick tests to fungi on 130 tobacco-processing workers and 112 control workers. Industrial hygiene survey and environmental mycological studies were also performed. The average dust concentrations ranged from 13.76 to 29.55 mg/m(3) in the tobacco processing workshops. Also, the numbers of fungi colonies in the processing environments were much higher than those in the control environments. The prevalences of chronic respiratory or nasal symptoms in exposed workers were significantly higher than those in control workers (p<0.05 or p<0.01). The prevalences of lung function abnormalities in the exposed workers were significantly higher than those in control workers (p<0.05). The positive prevalences of specific IgE reactions to fungi (26.92% for A.fumigatus and 51.54% for Rhizopus nigricans) in exposed workers were also significantly higher than those in control workers (p<0.01). The positive prevalences of the skin prick test showed that 18.46% of the exposed workers were positive to A. fumigatus and 23.85% were positive to Rhizopus nigricans. Our results suggested that tobacco processing workers may develop respiratory diseases related to tobacco dust and we consider that fungi might be one of the allergens causing allergic respiratory or nasal diseases in tobacco processing workers. PMID- 15824485 TI - Merkel cell carcinoma in patients with long-term ingestion of arsenic. AB - Merkel cell carcinoma (MCC) is a rare primary neuroendocrine carcinoma of the skin, mostly occurring late in life on sun-exposed body parts. Little is known about the specific etiological factors in the pathogenesis of MCC. A previous report indicated that arsenic exposure might cause MCC, which might be another specific type of skin cancer associated with arsenic exposure. On the southwest coast of Taiwan, high arsenic levels in artesian well water have been documented, and various diseases associated with arsenic exposure have been found to be prevalent in this area. We report two pathologically confirmed cases of MCC in patients who had histories of long-term ingestion of arsenic from drinking water. The tumors were on the anterior chest wall, an area less exposed to the sun, in both cases. The literature on the dose-response relationship between arsenic exposure and MCC is limited. We estimated that the total arsenic ingested by these two cases was around 14.7 and 2.6 gm, respectively. We also tried to assess the cancer risk on the basis of the estimated doses of arsenic exposure and the cancer risk model developed by the U.S. Environmental Protection Agency (EPA). The estimated lifetime target cancer risk was 1.3 x 10(-2) in Case 1 and 2.3 x 10(-3) in Case 2. Both are much higher than the 10(-6) upper limit on lifetime cancer risk put forth by the U.S. EPA health protection standard. We believe that arsenic intoxication played an important role in the carcinogenic process of MCC in our cases. PMID- 15824486 TI - Role of word-external contexts in native speakers' identification of vowel length in Japanese. AB - Five groups of native Japanese listeners were assigned to five conditions differing in word-external contexts. In the Intact condition, three types of target disyllables, /mVmV/, /mV:mV/, and /mVmV:/ were spoken in a carrier sentence at two speaking rates. In the other four conditions, the target disyllables of the Intact condition were excised from the original carrier sentence (Excised); embedded in the carrier sentence of the other rate (Mismatch); presented only with the preceding three syllables (Preceding), and presented only with the following three syllables (Following). The accuracy for identifying the word types was higher for the Intact than Excised and Mismatch conditions, indicating that the presence of the carrier sentence with an appropriate rate was important for accurate identification. It was also found that either the preceding or the following short phrase contained sufficient information for identifying the word types. Further, there were effects of the preceding and following phrases on both the first and second vowels of the targets, suggesting that the distant speech materials more than two phonemes away from a target in both sides can affect identification accuracy. Implications are discussed in terms of the adjacency principle for rate normalization. PMID- 15824487 TI - The stop voicing contrast in French sentences: contextual sensitivity of vowel duration, closure duration, voice onset time, stop release and closure voicing. AB - This study examined the manner in which French speakers used some acoustic correlates to produce the stop voicing distinction in French sentences when syllables containing syllable initial and -final stops were between vowels (/pa_a/) and between voiceless fricatives (/pas_s/). Data analyses revealed that /b, d, g/ were longer, were more frequently phonated, and were preceded by longer vowels than /p, t, k/ in three conditions: syllable-initial stops between vowels and between voiceless fricatives and syllable-final stops between vowels. When a voiceless fricative /s/ followed /b, d, g/, the voicing contrast was reduced as a result of complete regressive voicing assimilation, achieved by the concomitant devoicing of /b, d, g/ closures and the significant reduction in voicing-related differences in preceding vowel and closure durations. When /s/ preceded /b, d, g/, the voicing distinction was enhanced: significant voicing-related duration differences were accompanied by the complete assimilation of /s/ to [z]. Overall, findings suggest that in French sentences, voicing assimilation is strictly regressive and complete assimilation is achieved by the covariation of several acoustic correlates, which attests to the complementarity of the underlying articulatory gestures. PMID- 15824489 TI - Stroboscopic-cine MRI data on Korean coronal plosives and affricates: implications for their place of articulation as alveolar. AB - The present study aimed to explore the place of articulation of the Korean (pulmonic egressive) lenis, aspirated and fortis affricates /c, c(h), c'/ in comparison with that of the coronal plosive counterparts /t, t(h), t'/. For this purpose, a stroboscopiccine magnetic resonance imaging (MRI) experiment was conducted to examine not only where oral contact and constriction occur during oral closure and at its release, but also which part of the tongue is involved and how the apex, the blade and the body of the tongue are positioned as a function of time. Based on the results of the MRI experiment, we propose that the affricates should be classified as alveolar like the plosives but that they could be different in tongue shapes during oral closure and at its release: the affricates are laminal and the plosives are apical or apico-laminal. This distinction is associated with correlated variations in tongue body position (measured as pharyngeal width) and tongue blade position: tongue body position is more fronted with a relatively higher tongue blade position in affricates than in plosives. It is proposed that the gestural differences between the two types of stops serve to enhance the auditory contrast between the [+strident] affricates and the [-strident] plosives. PMID- 15824488 TI - Language-specific articulatory settings: evidence from inter-utterance rest position. AB - The possible existence of language-specific articulatory settings (underlying or default articulator positions) has long been discussed, but these have proven elusive to direct measurement. This paper presents two experiments using X-ray data of 5 English and 5 French subjects linking articulatory setting to speech rest position, which is measurable without segmental interference. Results of the first experiment show that speech rest position is significantly different across languages at 5 measurement locations in the vocal tract, and is similar to previously described language-specific articulatory settings. The second experiment shows that the accuracy of achievement of speech rest position is similar to that of a specified vowel target (/i/). These results have implications for the phonetics and phonology of neutral vowels, segmental inventories, and L2 acquisition. PMID- 15824490 TI - Prenatal diagnosis, phenotypic and obstetric characteristics of holoprosencephaly. AB - The diagnosis of fetal malformations, especially those of the central nervous system, is strikingly important in the practice of genetic counseling. Early diagnosis is very significant, not only because of the prognosis, but also because of the emotional effects caused by the accompanying craniofacial malformations. The summary of the obstetrical and diagnostical characteristics should be useful in the management of holoprosencephaly. The analysis of the 50 cases we encountered between 1981 and 2000, including the anatomical, diagnostic and clinical aspects, as well as the associated craniofacial malformations, forms the essence of our publication. In one of the examined cases a familiar recurrence was verified. PMID- 15824491 TI - Evaluation of a fetus with Neu-Laxova syndrome through prenatal, clinical, and pathological findings. AB - We report a case of Neu-Laxova syndrome in a fetus at 22 weeks with the ultrasonographic findings of characteristic facial findings, limb contractures, kyphosis and polyhydramnios. Pathological and ultrasonographic studies are discussed. PMID- 15824492 TI - Prenatal ultrasonographic diagnosis of rhizomelic chondrodysplasia punctata by detection of rhizomelic shortening and bilateral cataracts. AB - Antenatal sonographic diagnosis of rhizomelic chondrodysplasia punctata depends on recognization of the combination of rhizomelic bone shortening and epiphyseal stippling. This is the only report of prenatal ultrasonographic diagnosis of bilateral cataracts in a fetus with rhizomelic chondrodysplasia punctata (type 1). Also, this is the first report of severe rhizomelic limb shortening, and bilateral cataracts prior to the recognization of epiphyseal stippling. PMID- 15824493 TI - Heterogeneity in fetal immunocompetence during the second trimester of gestation. Implications for treatment of nonimmune genetic disorders by in utero transplantation. AB - OBJECTIVE: To address the role that alloreactivity may play and better define the window for histoincompatible stem cell transplantation in utero. SUBJECTS, MATERIAL AND METHODS: We studied 9 fetal blood specimens obtained by cardiocentesis during elective abortions in the second trimester by multicolor flow cytometry and in vitro stimulation. RESULTS: Lymphocytes ranged from adult levels (3/9) to >90% leukocytes. Six specimens had T cells within adult range. T cells in the other specimens were reduced, while B cells were conversely elevated. This variability did not correlate with gestational age, or leukocyte composition. Following 4 h of mitogenesis, fetal CD4+ and CD8+ T cells from 1 of 5 specimens showed a response similar to that of maternal T cells, while the other 4 specimens showed a diminished response (0.3 +/- 0.2-fold). This heterogeneity did not correlate with gestational age, or lymphocyte subset distribution. Following 18 h of in vitro mitogenesis, fetal T cells from 2 specimens showed a response similar to that of maternal T cells (0.8 +/- 0.2 fold). Despite that, one specimen gave a 3-fold greater response in a one-way mixed lymphocyte reaction vs. maternal cells compared to the other specimen. CONCLUSION: We determine that fetal immunocompetence differs greatly during the second trimester and assessment of host vs. donor reactivity prior to in utero transplantation is likely to potentiate more favorable outcomes. PMID- 15824494 TI - Oligohydramnios in uncomplicated pregnancies beyond 40 completed weeks. A prospective, randomised, pilot study on maternal and neonatal outcomes. AB - OBJECTIVES: In low-risk pregnancies, the management of oligohydramnios is not clear. The aim of this study was to compare maternal and neonatal outcomes in cases of isolated oligohydramnios randomised to either induction of labour or expectant management. METHODS: 87 women pregnant beyond 40 completed weeks were asked to participate in the study. Fifty-four accepted and were randomised at 288 days of pregnancy. Twenty-six were randomised to expectant management and 28 to induction of labour. The primary maternal outcome was the mode of delivery and the primary neonatal outcomes were cord blood pH and Apgar score at delivery. RESULTS: No significant differences were found for any important maternal or neonatal outcome. CONCLUSION: Based on these results, the decisions about management could be individualised. Even though the number of patients included was small, these results are well in concordance with others. PMID- 15824495 TI - Biochemical composition of amniotic fluid in pregnancies complicated with twin twin transfusion syndrome. AB - OBJECTIVE: To compare the electrolyte composition of pregnancies complicated with twin-twin transfusion syndrome (TTTS) with that of physiologic pregnancies. MATERIALS AND METHODS: Amniotic fluid samples from 16 pregnancies were studied. Specimens were obtained from recipient sacs in 10 pregnancies undergoing fetoscopy for severe midtrimester TTTS. Additionally, 6 amniotic fluid samples were obtained transcervically from legal second-trimester pregnancy terminations. The concentrations of sodium, potassium, chloride, bicarbonate, calcium, glucose, osmolality, pH, total protein content and albumin were determined in each sample. RESULTS: The mean gestational age at sampling was 20.2 weeks (range 17.2-27.1) in the TTTS group and 18.4 (range 16.0-22.0) in the control group (p = NS). We found significant lower levels of albumin (0.22 +/- 0.04 vs. 0.39 +/- 0.11, p = 0.01) and total protein (0.19 +/- 0.08 vs. 0.51 +/- 0.17, p < 0.001) and higher levels of bicarbonate (16.90 +/- 1.45 vs. 14.50 +/- 2.17, p = 0.02) in amniotic fluid samples taken from recipient sacs of TTTS pregnancies. CONCLUSION: Amniotic fluid from the receptor in severe midtrimester TTTS differs significantly from control amniotic fluid samples in bicarbonate concentration, total protein content and albumin concentration. These findings may help to understand the pathophysiology of TTTS and to optimise therapeutic modalities. PMID- 15824496 TI - Too late prenatal diagnosis of fetal toxoplasmosis: a case report. AB - OBJECTIVE: We describe a case of severe fetal hydrocephalus due to toxoplasmosis which could not be diagnosed until late gestational age due to the lack of a serologic surveillance program during pregnancy; moreover, this case points to the usefulness of molecular biology tools in the diagnostic process. Abnormal ultrasound in the 2nd trimester was noticed and Toxoplasma gondii was demonstrated in amniotic fluid at the 28th week of gestation both by PCR and by mice inoculation. Fansidar and folinic acid were administered. The newborn suffered from progressive hydrocephalus, seizures, and pathological muscular tonus; ultrasound examination showed massive cerebral calcifications. Ophthalmologic examination revealed bilateral choroidoretinitis. Congenital toxoplasmosis was confirmed by the detection of anti- T. gondii IgM and IgA in the neonatal serum. CONCLUSION: The presented case is an example of severe fetal toxoplasmosis diagnosed and treated in utero. PMID- 15824497 TI - Management for neonatal aspiration syndrome caused by vernix caseosa. AB - OBJECTIVE: To clarify resuscitation methods against the lethal vernix caseosa aspiration syndrome. METHOD: A single case report with a literature review. RESULTS: Our case report was associated with aspiration syndrome caused by an airway obstruction of vernix caseosa: a proteolipid biofilm synthesized by the fetus. A 23-year-old woman normally delivered a mature infant at term. The infantile oral cavity was filled with numerous aggregates of vernix caseosa. Two hours after his birth, the infant died from respiratory insufficiency. We discuss the difference between management of our case and the previously reported case. CONCLUSION: Pregnant women with a diffuse pattern of high-level echoes in prenatal ultrasonography, suggesting the presence of massive vernix caseosa, should be transferred to a well-equipped institution that can administer inhaled nitric oxide and extracorporeal membrane oxygenation. PMID- 15824498 TI - Antenatal presentation of isolated femoral hypoplasia discovered at 18 weeks of gestation. AB - Prenatal diagnosis performed by ultrasound scan is now a routine part of antenatal care in France. A case of congenital short femur in an otherwise healthy infant is described. Antenatal diagnosis was made at 18 weeks of gestation and ultrasonic follow-up was performed. The right femur was shorter than the left. Further sonographic exploration demonstrated unilateral femoral hypoplasia without another anomaly. In this case, the occurrence of congenital short femur was sporadic. The diagnosis was confirmed after delivery. Prenatal sonographic features, differential diagnosis, prognosis and management are discussed. This case illustrates the importance of ultrasound as an early detector of certain congenital anomalies and as a useful tool in their follow-up. PMID- 15824499 TI - Detection of viral deoxyribonucleic acid in amniotic fluid: association with fetal malformation and pregnancy abnormalities. AB - OBJECTIVE: To first test the hypothesis that the presence of viral nucleic acid in amniotic fluid (AF) is associated with an abnormal pregnancy outcome, and second, to determine if the overall rate of polymerase chain reaction (PCR) positivity and the distribution of virus types vary geographically. STUDY DESIGN: Cytomegalovirus (CMV), parvovirus B19, adenovirus, enterovirus, herpes simplex virus, Epstein-Barr virus, and respiratory syncytial virus nucleic acids were sought in 423 AF samples obtained for clinical indications: 284 from the East Coast (EC) and 139 from the Midwest (MW). RESULTS: Gestational age at sampling was 19.1 weeks for EC and 20.1 weeks for MW. 13.5% of karyotypically normal singleton pregnancies (57/423) had a positive AF PCR. 11% of AF PCR from the EC while 18% of AF PCR from the MW were positive (p = 0.06). The most commonly detected viruses were adenovirus (77%), enterovirus (12%), and CMV and parvovirus B19 (5% each). Twenty-four percent of sonographically abnormal pregnancies (33/136) had a positive AF PCR compared to only 8.4% (24/287) of normal pregnancies (p < 0.001). CONCLUSION: A positive AF PCR is associated with an increased rate of fetal structural malformations, intrauterine growth restriction, hydrops and other fetal abnormalities. There were no significant geographic differences in the incidence of AF viral PCR positivity. PMID- 15824500 TI - Epidural local anesthetics: a novel treatment for fetal growth retardation? AB - BACKGROUND: Chronically compromised uterine perfusion may lead to placental insufficiency and subsequent intrauterine growth restriction (IUGR). Various therapeutic approaches (e.g. vasodilators, low-dose aspirin, intravenous glucose infusion, and hemodilution) are often of limited efficacy. Local anesthetics have been shown to improve placental blood flow in pre-eclamptic women. We hypothesized that epidural administration of local anesthetics might improve outcome in IUGR independent of the underlying cause. In preparation for a clinical trial to test this hypothesis, we performed a pilot study in 10 patients. METHODS: After approval of the study protocol, 10 pregnant women presenting with oligohydramnios and IUGR were included in the study. In addition to our standard protocol (magnesium, glucose, betamethasone), each patient received an epidural catheter (T10/T12) with continuous infusion of bupivacaine 0.175% at a rate of 5 ml/h. Uteroplacental circulation was monitored by Doppler sonography and the amount of amniotic fluid was estimated daily. RESULTS: Epidural insertion and infusion was performed without complications. Four patients continued to deteriorate rapidly, amniotic fluid volume did not change and uterine artery pulsatility index (PI) tended to increase. In the remaining 6 patients the clinical status stabilized, amniotic fluid volume tended to increase and uterine artery PI tended to decrease during treatment. This improvement was associated with a prolonged interval to cesarean section and increased infant birth weight. CONCLUSION: Our data suggest that, even if the underlying cause of IUGR is not pre-eclampsia, epidural local anesthetic administration might improve placental blood flow and be beneficial in a subgroup of patients. A clinical trial to test this hypothesis appears warranted. PMID- 15824501 TI - The ex utero intrapartum treatment (EXIT) procedure in giant fetal neck masses. AB - Giant fetal neck masses can cause airway obstructions with potential poor fetal prognosis after delivery. The relationship between the fetal neck mass and airway structure can be defined prenatally with ultrasound and magnetic resonance imaging (MRI). The ex utero intrapartum treatment (EXIT) procedure is an available technique to obtain a fetal airway while feto-maternal circulation is preserved. We present a case in which prenatally a giant fetal neck mass was diagnosed on ultrasound and MRI, and a successful EXIT procedure could be performed. PMID- 15824502 TI - Tetrasomy 9p mosaicism associated with a normal phenotype. AB - Isochromosome (tetrasomy) 9p is a rare chromosomal aberration characterized by phenotypic abnormalities ranging from mild developmental delay to multiple anomalies including intrauterine growth retardation, cerebral ventriculomegaly, dysmorphic facial features, cleft lip or palate, abnormal genitalia and renal anomalies. We present a patient with isochromosome (tetrasomy) 9p mosaicism who is a healthy normal adult male with oligospermia who has fathered two normal children. This chromosomal abnormality may be tissue specific, with a higher detection rate in cultured lymphocytes compared with fibroblasts. Therefore, there is an increased chance of missing the abnormality prenatally by amniocentesis or chorionic villus sampling. We are aware of only one other patient in the literature with a normal phenotype associated with mosaicism for this chromosomal abnormality. PMID- 15824503 TI - The ethics of gene therapy and abortion: public opinion. AB - On the grounds that the public should be consulted in decisions concerning the legitimate scope of germ-line genetic therapy (GLGT), survey data on the ethics of GLGT were collected from a large (n = 1,403) representative national sample of Australians in 2002. The data show that opinion is quite divided over GLGT in the case of a 'death sentence' genetic defect: 36% would forbid it, 23% have mixed feelings and 41% would allow it. For less serious conditions there is more opposition to GLGT. Thus, 48% would forbid GLGT to remedy a minor physical defect and 52% would oppose GLGT to counteract a propensity to violence, but fully 73% would disallow GLGT for cosmetic reasons. The data also show that opposition to abortion is lower than opposition to GLGT in the case of a 'death sentence' genetic defect, but at about the same level as, or greater than, opposition to GLGT for less serious issues. The questions show good measurement properties, including low missing data rates, so they are likely to provide an accurate picture of the public's views on the ethics of GLGT. It is suggested that a system for monitoring public opinion on these issues be developed. PMID- 15824504 TI - 2nd-trimester maternal serum human chorionic gonadotropin and alpha-fetoprotein levels in male and female fetuses with Down syndrome. AB - BACKGROUND/OBJECTIVE: Several studies have shown that the 2nd-trimester maternal serum alpha-fetoprotein (AFP) level is significantly lower and that the maternal serum human chorionic gonadotropin (hCG) level is significantly higher in the presence of a female fetus. This may potentially affect Down syndrome (DS) screening such that a higher false-positive rate may occur in women carrying a female fetus, whereas a lower detection rate may result in those carrying males. The purpose of this study was to evaluate the gender impact on marker levels in DS pregnancies and its effect on DS screening. METHODS: The study included 62 DS pregnancies with a single fetus of known gender (31 male and 31 female). Only pregnancies with chromosomal analysis showing trisomy 21 were included. The maternal serum levels of hCG, AFP, and unconjugated estriol were measured at 16 20 weeks of pregnancy. These levels were expressed as gestational-age-corrected multiples of the median. RESULTS: No statistically significant differences were noted in maternal serum levels of hCG or AFP in DS pregnancies between women carrying a female and those carrying a male DS fetus. No statistically significant differences in 'screen-negative' rates were noted among male and female fetuses. CONCLUSIONS: In normal pregnancies, the maternal serum hCG level is higher, and the AFP level is lower in the presence of a female fetus. However, this gender-related difference is not apparent in DS pregnancies. Therefore, the gender-related differences in serum marker levels would not result in a lower detection rate of DS in male fetuses. PMID- 15824505 TI - Prenatal screening for Down syndrome: didn't we forget something? PMID- 15824506 TI - MMF for lupus nephritis as a possible first aid block. PMID- 15824507 TI - Dysregulation of superoxide dismutase in chronic kidney disease. PMID- 15824508 TI - Comparison of intravenous iron sucrose to oral iron in the treatment of anemic patients with chronic kidney disease not on dialysis. AB - BACKGROUND: Few studies compare oral to intravenous (IV) iron for managing anemia in patients with chronic kidney disease (CKD) not on dialysis. METHODS: We enrolled 96 CKD anemic patients on erythropoietin in a randomized, open-label, multicenter, controlled study. Patients received 29 days of oral FeSO4 (325 mg t.i.d.) or intravenous (IV) iron sucrose (5 doses of 200 mg weekly). Assessments were made up to 14 days after the last dose. Primary endpoints were changes in hemoglobin and ferritin, and clinical success was evaluated from the percent of patients with combined endpoints of rises in hemoglobin/ferritin, hemoglobin/ferritin/TSAT, and hemoglobin/TSAT. RESULTS: There was no significant difference in hemoglobin values between IV and oral therapy. IV iron patients had greater increases in mean serum ferritin (288 ng/ml, p < 0.0001) compared to oral iron patients (-5.1 ng/ml, p = NS). IV iron patients with baseline ferritin < 100 ng/ml had a greater increase in hemoglobin (1.4 g/dl) compared to oral iron patients (0.9 g/dl) (p < 0.05). More IV iron patients (54.2%) attained hemoglobin values > 11.0 g/dl compared to oral iron patients (31.3%, p = 0.028), and met hemoglobin/ferritin (62.5%), hemoglobin/TSAT (47.9%), hemoglobin/ferritin/TSAT (43.8%), and ferritin/TSAT criteria (54.2%) than oral iron patients (0, 22.9, 0, and 0%, respectively). There were no serious side effects. CONCLUSIONS: These CKD patients had increases in both hemoglobin and ferritin following IV iron therapy, whereas those treated with oral iron had increases in hemoglobin without increases in iron stores. Iron sucrose, given weekly as 200 mg IV push over 5 min is an effective and safe anemia treatment in this population. PMID- 15824509 TI - Gene expression of perforin by peripheral blood lymphocytes as a marker of acute rejection. AB - BACKGROUND: Previous findings have demonstrated that the expression of cytotoxic effector molecules is increased in acute rejection of renal allografts. In the present study, we serially examined the gene expression of perforin, granzyme B and Fas ligand (FasL) in peripheral blood lymphocytes (PBLs) of renal allograft recipients to assess the potential of their expression as a marker of acute rejection. METHODS: PBLs were isolated from blood samples taken on days 2, 4, 6, 8, 10 and 12 after transplantation. Competitive PCR was performed to evaluate the abundance of mRNA of perforin, granzyme B and FasL. The mean value + 2 SD of each molecule in the control group was set as a discriminatory level for that particular molecule. RESULTS: When all measured samples were compared, perforin expression was significantly higher in patients with acute rejection than in the control group (1.84 +/- 3.01 vs. 0.71 +/- 0.48, p = 0.01). The percentage of perforin expression exceeding the discriminatory level was also significantly higher in patients with acute rejection (p = 0.0003). Five patients in the rejection group (5/7, 71.4%) showed perforin expression exceeding the discriminatory level, while only 1 patient in the control group did so (1/8, 12.5%) (p = 0.02). Perforin expressions of days 0 and 1 of rejection crisis were the highest over the study period. No consistent pattern of granzyme B and FasL expression was identified in relation to rejection crisis. CONCLUSION: Gene expression of perforin by PBLs was upregulated in accordance with acute rejection, thus offering the possibility that it may be utilized as a marker of acute rejection. PMID- 15824510 TI - Lymphocyte apoptosis: role of uremia and permeability of dialysis membrane. AB - BACKGROUND: Uremia is associated to host defense mechanism disorders. Lymphocyte apoptosis, which may cause alteration of the immune system, is increased in uremic patients. The aim of the present study was to determine if, in addition to uremia, dialysis membranes with different biocompatibility and permeability have an effect on lymphocyte apoptosis. METHODS: Cell apoptosis and Fas expression were assessed using flow cytometry in four groups of patients: (1) uremic non dialyzed (Non-D) patients; (2) hemodialysis (HD) patients on hemophan; (3) low flux polysulfone, and (4) high-flux polysulfone membrane. Ten healthy volunteers were used as controls. RESULTS: At baseline, lymphocytes from patients on hemophan showed an increase in apoptosis (18.4 +/- 6.9%) as compared with Non-D (7.2 +/- 2.8%; p < 0.001), low-flux (6.4 +/- 2.4%; p < 0.001), high-flux (2.6 +/- 1.2%; p < 0.001) and controls (2.0 +/- 1.0%; p < 0.001). Fas expression was similar in lymphocytes from Non-D and hemophan dialyzed patients (40.5 +/- 5% vs. 40.4 +/- 6%), and in both groups it was greater than low-flux (30%+/-7%; p < 0.001), high-flux (11 +/- 4%; p < 0.001) and controls (12.6 +/- 3%; p < 0.001). When lymphocytes were cultured for 48 h, apoptosis was similar in Non-D and hemophan (27.0 +/- 4.3% vs. 27.1 +/- 6.9%); apoptosis of lymphocyte from patients on low-flux (14.1 +/- 3.5%) was greater than on high-flux polysulfone membrane (7.0 +/- 2.0%; p < 0.001). CONCLUSION: These findings suggest that in dialysis patients lymphocyte apoptosis is influenced not only by the biocompatibility but also by the permeability of the dialysis membrane. PMID- 15824511 TI - Efficacy and safety of intravenous iron therapy for HCV-positive haemodialysis patients. AB - BACKGROUND: Iron supplementation is the cornerstone of anaemia management in haemodialysis (HD) patients. However, efficacy and safety of intravenous (IV) iron therapy in hepatitis C virus (HCV)-positive HD patients is yet to be elucidated. METHODS: Sixty-six maintenance HD patients with suboptimal response to recombinant human erythropoietin (rh-EPO) were administered IV iron. Each patient received 100 mg/session IV iron sucrose for ten consecutive HD sessions and then the dose was decreased to 50-100 mg weekly or biweekly. Patients were followed for haemoglobin (Hb), ferritin, rh-EPO dose requirements, transaminase levels, and adverse drug reactions. RESULTS: Baseline demographic and clinical characteristics, as well as Hb, ferritin, transaminase levels, rh-EPO and iron doses were similar between HCV-positive (n = 32) and HCV-negative patients (n = 29). After 5 months of follow-up, a significant increase in ferritin and Hb levels and decrease in rh-EPO doses were observed in both groups. The incidence of adverse drug reactions was not associated with HCV serology. Significant elevation in both alanine and aspartate aminotransferase levels were detected in HCV-positive patients. CONCLUSION: This study has shown that IV iron administration reverses suboptimal response to rh-EPO administration in HD patients regardless of HCV serology. There is however subtle increase of transaminase levels in HCV-positive patients. Further studies are warranted to reveal the impact of variation in serum transaminase levels during IV iron administration in HCV-positive HD patients. PMID- 15824512 TI - Renal involvement of non-Hodgkin's lymphoma and its prognostic effect in childhood. AB - OBJECTIVE: To evaluate renal involvement in childhood lymphoma and define its prognostic effects. PATIENTS AND METHODS: One hundred and four patients with non Hodgkin's lymphoma and renal involvement on admission to a single center between 1972 and 2003 were evaluated retrospectively. Blood urea nitrogen, serum creatinine, uric acid, electrolytes, and lactate dehydrogenase levels, as well as urinalysis, were evaluated. One or more of the following imaging methods were performed: intravenous urogram, ultrasound, computed tomography, and magnetic resonance imaging. The chi2 test was used to compare the groups. The Kaplan-Meier survival method was used to calculate survival rates, and the log-rank test was used to compare groups with respect to survival. Survival rates were also compared in two different time periods (before 1991 and after 1991). RESULTS: There were 76 boys and 28 girls with a median age of 6 (0.9-16) years. The renal infiltration pattern was nodular in 62 patients (59.6%) and diffuse in 40 patients (38.5%). Two patients had tumoral masses that originated from their kidneys (1.9%). Renal involvement was bilateral in 75 patients (72.1%); the remaining 29 patients had unilateral involvement. The overall survival rate was 42.5% with a median follow-up of 64 months. The factors that had a statistically significant impact on survival were high creatinine (p = 0.00001) and blood urea nitrogen levels (p = 0.0001), the onset of tumor lysis syndrome (p = 0.01), and the need for dialysis (p = 0.009). The survival rate was higher in the time period after 1991 (p = 0.01). CONCLUSION: Impaired renal function is a poor prognostic factor for non-Hodgkin's lymphoma. Renal function should therefore be monitored closely. Renal dysfunction caused by direct tumoral involvement may complicate therapy and shorten survival. PMID- 15824513 TI - Mycophenolate mofetil for remission induction in severe lupus nephritis. AB - BACKGROUND: Mycophenolate mofetil (MMF) is a potential alternative immunosuppressive to cyclophosphamide or azathioprine for the treatment of lupus nephritis. It has a superior toxicity profile to cyclophosphamide and is more effective than azathioprine when used in combination with cyclosporin for renal transplantation. METHODS: This open label study assessed the safety and efficacy of an induction regimen of MMF and prednisolone in 24 patients, with active WHO Class III, IV or V lupus nephritis, without previous exposure to cyclophosphamide or MMF. Patients received MMF 2 g/day and a tapering dose of oral prednisolone and were followed for 12 months. Renal response was defined using the validated BILAG (British Isles Lupus Assessment Group) Index. RESULTS: 20 patients achieved complete renal remission, 2 partial renal remission and 2 had renal disease refractory to MMF. Five patients were withdrawn, 2 for progressive renal disease, 2 for extra-renal flares and 1 following a severe infection. No patient was withdrawn for drug intolerance. There were 18 adverse events of which 10 were infections. Clinical improvement was mirrored by normalisation of complement levels and marked falls in circulating anti-double-stranded DNA antibodies. Follow-up for a mean period of 35 months documented only one episode of renal relapse which subsequently remitted with re-introduction of MMF. One patient died after 18 months' follow-up with uterine malignancy. CONCLUSIONS: MMF with prednisolone was effective for remission induction in severe lupus nephritis and was well tolerated. PMID- 15824514 TI - A2A adenosine receptor agonist and PDE4 inhibition delays inflammation but fails to reduce injury in experimental obstructive nephropathy. AB - BACKGROUND: Renal interstitial inflammation is a consequence of unilateral ureteral obstruction (UUO). Following ischemia/reperfusion, adenosine reduces renal inflammation and injury, effects which are potentiated by type 4 phosphodiesterase inhibitors. We therefore studied the effects of A2A adenosine receptor agonist (ATL146e), and PDE4 inhibitor (rolipram) in mice subjected to UUO. METHODS: Mice were subjected to UUO or sham operation, and received either vehicle or ATL146e + rolipram by osmotic minipump for 1 or 7 days. At 1, 3, 7, or 14 days after operation, renal macrophage infiltration, apoptosis, proliferation, tubular atrophy, and interstitial fibrosis were quantitated, and expressions of IL-6 and TGF-beta mRNA were determined. RESULTS: ATL146e + rolipram reduced macrophage infiltration by 40% after 3 days UUO (p < 0.05). Tubular apoptosis, tubular atrophy, and interstitial fibrosis were increased by 7 or 14 days UUO, but were unaffected by ATL146e + rolipram. However, cellular proliferation was increased by ATL146e + rolipram in the obstructed kidney. ATL146e + rolipram had no effect on the renal expression of IL-6 and TGF-beta mRNA. CONCLUSIONS: A2A receptor activation and PDE4 inhibition transiently reduce renal macrophage infiltration, but do not ameliorate the renal response to UUO. We speculate that the persistent stimulus for inflammation triggered by UUO cannot be reversed by agents that suppress inflammatory cell activation alone. PMID- 15824515 TI - STAT1-independent down-regulation of interferon-gamma-induced class II transactivator and HLA-DR expression by transforming growth factor beta-1 in human glomerular endothelial cells. AB - BACKGROUND: The competition between STAT1 and Smad3 for a limiting amount of the nuclear protein p300, a transcriptional coactivator, was suggested to be a mechanism for the antagonism between interferon-gamma (IFN-gamma) and transforming growth factor-beta1 (TGF-beta1). We investigated the effect of TGF beta1 on IFN-gamma-induced HLA-DR production in cultured human glomerular endothelial cells (HGECs), and the involvement of p300 in this process. METHODS: Cell surface expression of HLA-DR and mRNA levels of HLA-DR and class II transactivator (CIITA), the master regulator of HLA-DR gene transcription, were measured by cellular ELISA and Northern blot, respectively. The levels of STAT1 and Smad3 protein were analyzed by Western blot. Nuclear binding activity of STAT1 was assessed by electrophoretic mobility shift assay. RESULTS: IFN-gamma increased the cell surface expression of HLA-DR along with increases in the mRNA levels of CIITA and HLA-DR, while these stimulatory effects of IFN-gamma were down-regulated by TGF-beta1. IFN-gamma increased phosphorylation of STAT1 and this activation was not inhibited by TGF-beta1. IFN-gamma increased binding of p STAT1 to p300, while TGF-beta1 increased binding of Smad3 to p300. TGF-beta1 induced Smad3 binding to p300 was inhibited by IFN-gamma, whereas IFN-gamma induced p-STAT1 binding to p300 was not inhibited by TGF-beta1. IFN-gamma increased DNA binding activity of STAT1. Inhibition of interaction between STAT1 and p300 by addition of anti-p300 antibody to nuclear extract down-regulated DNA binding activity of STAT1. In contrast, TGF-beta1 did not inhibit IFN-gamma induced STAT1 binding to DNA. CONCLUSIONS: TGF-beta1 down-regulated IFN-gamma induced CIITA and HLA-DR expression in HGECs. Though there was an antagonism between IFN-gamma and TGF-beta1, the competition for p300 between p-STAT1 and Smad3 was not the mechanism for it. PMID- 15824516 TI - Suppressive mechanisms of Sairei-to on mesangial matrix expansion in rat mesangioproliferative glomerulonephritis. AB - BACKGROUND: Sairei-to (TJ-114) is a Japanese herbal medicine of standardized quality, originating from traditional Chinese medicine. In the present in vivo study, we investigated the suppressive effects of TJ-114 and related drugs, Shosaiko-to (TJ-9), and Saiboku-to (TJ-96), on mesangioproliferative glomerulonephritis (MsPGN) in rats. TJ-9 is a basal prescription of TJ-96 and TJ 114. We evaluated the efficacy of these drugs on proteinuria, extracellular matrix (ECM) accumulation, and superoxide dismutase (SOD)-activity. METHODS: MsPGN in Wistar rats was induced by intravenous injection of rabbit anti-rat thymocyte serum (ATS). TJ-114, TJ-9, TJ-96 (500 mg/kg/day), or prednisolone (PSL, 2 mg/kg/day) was orally administered to the rats as drinking water from the day of ATS injection (day 0) to day 8, when rats were sacrificed and the kidney specimens were collected. Macrophage infiltration was evaluated by immunostaining for ED-1. ECM was measured by trichrome-staining, and fibronectin immunostaining. Northern blotting was performed to clarify the mRNA expression of cytokines and fibronectin. SOD-activity in the homogenate of renal cortex was also evaluated. RESULTS: The amount of urinary protein was significantly decreased only in the TJ 114-treated group compared with the disease control group (p < 0.05). The number of ED-1-positive cells was significantly decreased in all the treatment groups (p < 0.05, respectively). Decreases in the trichrome-stained area were observed moderately in the TJ-114-treated group (66% of control, p < 0.001) and mildly in the PSL-treated group (76% of control, p < 0.001). The staining area of fibronectin in the glomerulus was significantly decreased in all the treated groups except PSL, and was especially suppressed in the TJ-114-treated group (45% of control, p < 0.001). Transforming growth factor (TGF) and connective tissue growth factor (CTGF) expression significantly decreased in the TJ-114-treated group to the control level (p < 0.05). TGF-beta, CTGF, and fibronectin mRNA were upregulated in the disease control group, and TJ-114 suppressed these mRNA expressions in glomeruli. The SOD-activity of renal cortex-homogenate was significantly augmented in all the treated groups except PSL, markedly in the TJ 96- and TJ-114-treated groups. CONCLUSION: These results suggest that TJ-114 ameliorates ECM accumulation in experimental rat MsPGN, partly suppressing TGF beta and CTGF expression through the recovery of SOD-activity. PMID- 15824517 TI - A correlation study of endoneurial fluid pressure and electroneurography of the facial nerve. AB - OBJECTIVE: The relationship between endoneurial fluid pressure (EFP) and electroneurography (ENoG) of the facial nerve was studied in order to evaluate the pathophysilogical basis of ENoG to serve as the criteria for decompression of the facial nerve. METHODS: While the values of ENoG were recorded by an instrument for physiology on the normal and crushed facial nerves of guinea pigs, EFPs were measured at the same time by a servo-nulling micropipette system. After the elevated EFP was measured, the facial nerves were removed and then fixed properly for examination under the light and electron microscope in order to determine the differences from various periods after injury. RESULTS: With the change of EFP in the facial nerve during the period from day 3 to the third week after crushed injury, the percentage of degenerated facial nerve fiber increased. There was a positive correlation between ENoG and EFP. The coefficient was 0.88 and 0.51 in the second and third week after crushed injury, respectively. Extensive edema in endoneurial and perivascular spaces of the facial nerve could be found at the early stage of injury, while proliferation of numerous Schwann cells appeared at the later stage. CONCLUSION: The change of ENoG could reflect the value of EFP relatively within 3 weeks after crushed injury. Our data indicate that ENoG could be useful to evaluate the pathogenesis underlying facial palsy. PMID- 15824518 TI - Inflammation--a new therapeutic target in pneumonia. AB - Inflammation is a hallmark of pneumonia. Therefore, managing inflammation is an attractive adjunct to targeted antibiotic therapy, mainly in severe pneumonia. Recent investigations indicate that glucocorticoids given in physiological doses (from 10-fold to 100-fold less than doses administered in the past) could be of benefit. We could also manage inflammation by administering or influencing cytokines. A major concern is that drugs designed to target a single cytokine or receptor could prove ineffective due to the redundancy of signaling pathways involved. This may require selection of drugs with broad activity or the targeting of molecules common to inflammatory signaling pathways. Drugs affecting multiple molecules or key inflammatory pathway intermediates could be more effective, but their use will need to be weighed against the risk of impairing innate immunity. Indirect approaches to manage inflammation, such as neutralizing cytotoxic substances in the lung (e.g., inhibiting, neutralizing and eliminating endotoxin), could be used in combination with other approaches. Ideally, potential treatment of life-threatening bacterial pneumonia will combine immunoadjuvant and conventional antibiotic therapy, although intense clinical research with immunotherapy has not yet yielded a successful treatment adjunct. We believe that compounds capable of stimulating early host defense and microbial clearance, but not the later phases of inflammatory tissue injury associated with sepsis, may be advantageous. PMID- 15824519 TI - Improving the evaluation of diffuse infiltrative lung diseases by means of mutual education: difficult task, but worth a try. PMID- 15824520 TI - It is time for this 'ROSE' to flower. PMID- 15824521 TI - It's not as simple as it seems: continuous positive airway pressure and cheyne stokes respiration. PMID- 15824522 TI - Do high-resolution CT findings of usual interstitial pneumonitis obviate lung biopsy? Views of pulmonologists. AB - BACKGROUND: High-resolution CT (HRCT) of the lungs has become an essential component to evaluate patients with diffuse lung disease. Little is known, however, about the current practices of pulmonologists caring for patients with these complex conditions, and, in particular, whether HRCT can obviate the need for surgical lung biopsy. OBJECTIVES: To investigate the practices of pulmonologists concerning the acceptability of a HRCT diagnosis in lieu of lung biopsy in diffuse lung disease. METHODS: We asked practicing pulmonologists among membership of the American College of Chest Physicians whether HRCT results could replace lung biopsy in 16 diffuse lung diseases. Responses were examined in light of published evidence, practice guidelines, and certain practice parameters. RESULTS: Two hundred and thirty (52.6%) of 437 eligible physicians responded. Sixty-seven percent (67%) of respondents accepted HRCT diagnosis for idiopathic pulmonary fibrosis/usual interstitial pneumonia (IPF/UIP) despite their awareness of guidelines recommending histological diagnosis. Most would not accept a radiologic diagnosis for lymphangioleiomyomatosis (LAM; 37%) or eosinophilic granuloma (Langerhans' cell histiocytosis, LCH; 19%), even though CT findings are frequently characteristic. Responses were similar by type of clinical practice and recency of fellowship training. Chest physicians who referred patients for HRCT more frequently were more likely to accept HRCT diagnosis (p=0.008) and those who had higher self-ratings of proficiency in reading HRCT (p = 0.004) were more likely to believe HRCT often suggests specific diagnosis. CONCLUSIONS: Most US pulmonologists will accept an HRCT diagnosis of IPF/UIP without lung biopsy, but are reluctant to do so for most other diffuse lung conditions including LAM and LCH. PMID- 15824524 TI - Acute effects of automated continuous positive airway pressure on blood pressure in patients with sleep apnea and hypertension. AB - BACKGROUND: The obstructive sleep apnea-hypopnea syndrome (OSAHS) is characterized by repetitive upper airway obstructions during sleep, and it might cause cardiovascular complications such as myocardial infarction, arrhythmias, and systemic and pulmonary hypertension. OBJECTIVES: We investigated the acute effects of automatic continuous positive airway pressure (automated CPAP) on blood pressure in patients with OSAHS and hypertension. METHODS: Polysomnography was used and ambulatory blood pressure measurements were done in 12 patients with OSAHS. Blood pressure and heart rate were measured at night (10 p.m. to 6 a.m.) and during the day (6 a.m. to 10 p.m.). During these periods systolic, diastolic and mean blood pressure and heart rate of the patients on the diagnostic day were compared with those on the treatment day. RESULTS: Patients had moderate or severe OSAHS; their mean age was 52.8+/-4.2 years. Systolic, diastolic and mean blood pressure and heart rate between the diagnostic and treatment day were not significantly different. Standard deviations of all these parameters during the night of the treatment day (9.1+/-4.5, 7.5+/-3.3, 8.0+/-3.0 mm Hg, and 4.8+/-1.5 beats/min, respectively) were significantly lower than during the night of the diagnostic day (12.6+/-4.9 mm Hg, p=0.023, 10.8+/-3.5 mm Hg, p=0.004, 11.6+/-4.4 mm Hg, p=0.006 and 6.9+/-1.6 beats/min, p=0.003, respectively). We did not find similar results during daytime periods. CONCLUSIONS: Automated CPAP therapy in patients with sleep apnea and hypertension did not decrease systolic and diastolic blood pressures and heart rates acutely. However, it might reduce the variability of these parameters during sleep in patients, but not during the day. It might be suggested that automated CPAP reduces cardiovascular morbidity of OSAHS via stabilizing heart rate and blood pressure during sleep. PMID- 15824523 TI - There is no relationship between chronic obstructive pulmonary disease and obstructive sleep apnea syndrome: a population study. AB - BACKGROUND: Both chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea syndrome (OSAS) are common diseases. Some recent studies suggest an increased prevalence of COPD among subjects with OSAS. OBJECTIVES: The study objective was to evaluate whether there is an epidemiological relationship between COPD and OSAS in a random population sample. MATERIALS AND METHODS: The study population, 356 males (53%) and 320 females, mean age 56.6+/-8.2 years (range 41-72), was selected from a voting list for parliamentary election in Warsaw. The investigation included lung diseases and smoking history with polysomnography and spirometry. RESULTS: OSAS was diagnosed in 76 subjects (11.3%), 59 males (8.8%) and 17 females (2.5%), mean apnea/hypopnea index (AHI) was 25.3+/-16.1, mean overnight SaO2 92.1+/-3.3%, minimum SaO2 76.9+/-9.4%, and SaO2<90%=18.9+/-23.9% of total sleep time. COPD was diagnosed in 72 subjects (10.7%), 39 males and 33 females. Severity of airflow limitation was assessed according to European Respiratory Society (ERS) guidelines: mild in 70%, moderate in 22%, and severe in 8%. In 7 subjects (9.2% of OSAS population, 1% of total population) OSAS and COPD overlapped. Polysomnographic variables were compared between overlap (overlap syndrome, OS) and OSAS subjects. In the OS mean AHI was 19.0 versus 25.3 in OSAS (nonsignificant), mean SaO2 89.6 versus 92.3% in OSAS (p<0.005), and time spent in SaO2<90% was 25.4 versus 18.2% in OSAS (p=0.04). CONCLUSIONS: COPD in subjects with OSAS was as frequent as in the general population. In the OS group mean arterial blood saturation was lower and time spent in desaturation was longer than in OSAS. The presented data suggest a more severe course of sleep-disordered breathing in subjects with coexisting COPD. PMID- 15824525 TI - A new parameter in the detection of tuberculosis activity: reactive oxygen metabolites. AB - BACKGROUND AND OBJECTIVES: In countries with a high frequency of tuberculosis, there are problems not only with active lung tuberculosis but also with past lung tuberculosis. Cases with sequel tuberculosis very frequently present with complaints like tuberculosis, and it is very hard to determine whether it is a sequel tuberculosis complication or reactivation of tuberculosis. In this study, we measured the serum reactive oxygen metabolite (ROM) levels of patients with active pulmonary tuberculosis and healthy controls, and investigated if these metabolites can be used as a criterion for differentiation between active pulmonary tuberculosis and sequel pulmonary tuberculosis. METHODS: 40 patients with active tuberculosis, 35 patients with sequel pulmonary tuberculosis and 30 healthy control subjects with a similar age range and sex distribution were included in the study. Serum total ROM levels were detected in the patients and control group. RESULTS: Mean serum ROM values were 994+/-236, 551+/-135 and 236+/ 59 U/l among active lung tuberculosis cases, sequel lung tuberculosis cases and the healthy control group, respectively. As a result of these findings, serum ROM levels of active lung tuberculosis cases and sequel lung tuberculosis cases were significantly higher than those of the control group (both p<0.001). The serum ROM levels of active lung tuberculosis cases were also significantly higher than those of sequel lung tuberculosis cases (p<0.001). CONCLUSIONS: In the light of our findings, it may be assumed that serum total ROM values can be used as an activity criterion in the differentiation of active lung tuberculosis and sequel lung tuberculosis. PMID- 15824526 TI - Tuberculous effusion: ADA activity correlates with CD4+ cell numbers in the fluid and the pleura. AB - BACKGROUND: Adenosine deaminase (ADA) is a commonly used marker in the diagnosis of tuberculous effusion and there is evidence that its production is linked to T cells and monocytes. Data on the correlation between ADA and T cells or macrophages in tuberculous effusions are conflicting. Furthermore, no studies have examined a possible correlation between pleural tissue infiltration and ADA. OBJECTIVES: We undertook this study to examine cell subsets in the fluid and the pleura in tuberculous effusion and their correlation to ADA. The use of cell subsets as a marker in the differential diagnosis was also examined. METHODS: Pleural fluid from 36 patients with tuberculous and 34 patients with malignant effusion as well as pleural tissue biopsies from 16 patients with tuberculous pleurisy were examined. The APAAP and the avidin-biotin complex immunocytochemical methods were used to examine CD4+ T cells and macrophages (CD68+), while ADA activity was measured by the Giusti colorimetric method. RESULTS: Our results showed that, in pleural fluid, CD4+ cells and ADA were significantly higher in tuberculous compared to malignant effusion (p<0.001 for all measurements). In pleural tissue biopsies, macrophages were the predominant cells but CD4+ T cells were also abundant. A significant correlation was found between ADA and CD4+ numbers in pleural fluid and tissue (r=0.45, p<0.01; r=0.75, p<0.001, respectively). ADA had high sensitivity and specificity for differential diagnosis while cell subsets did not. CONCLUSIONS: These results indicate that ADA activity correlates to CD4+ T cell infiltration in the pleura and the fluid. Moreover, ADA but no cell subsets may be used as markers of tuberculous effusion. PMID- 15824527 TI - Expression of matrix metalloproteinase-9 in pleural effusions of tuberculosis and lung cancer. AB - BACKGROUND: Matrix metalloproteinase (MMP)-9 has been implicated in the development of pleural effusions. OBJECTIVES: The aim of this study was to assess the expression of MMP-9 in pleural effusions of tuberculosis, lung cancer and transudates. METHODS: Ninety-one patients (37 tuberculous pleural effusions, 42 malignant pleural effusions of lung cancer and 12 transudates) were included. Concentrations of pleural fluid MMP-9 and tissue inhibitors of matrix metalloproteinase (TIMP)-1 were determined by immunoassay. We also investigated the cellular localization of MMP-9 and TIMP-1 by reverse-transcriptase polymerase chain reaction on lymphocytes from pleural effusions and by immunohistochemical analysis of pleural tissues. RESULTS: Pleural fluid MMP-9 levels, MMP-9/total protein and MMP-9/TIMP-1 ratios were significantly higher in tuberculous pleural effusions, whilst TIMP-1 levels were similar in the three groups. MMP-9 levels positively correlated with TIMP-1 and lactate dehydrogenase levels, and negatively with pH and glucose levels in pleural effusions. MMP-9 mRNA expression in lymphocytes tended to be higher in malignant pleural effusions of lung cancer than in the other groups without reaching statistical significance. The strongest immunoreactivity for MMP-9 was observed in epithelioid cells of tuberculous pleural tissues. Much lower levels of MMP-9 expression were found in tumor cells of pleural tissues. CONCLUSIONS: MMP-9 is increased in tuberculous pleural effusions compared with transudates and malignant pleural effusions of lung cancer and is produced predominantly by epithelioid cells in the granulomas of tuberculous pleural tissues. PMID- 15824528 TI - Blood gas analysis and chest x-ray findings in infants and preschool children with acute airway obstruction. AB - BACKGROUND: The importance of SaO2 in the assessment of respiratory distress in bronchial asthma has been reported. OBJECTIVES: To evaluate the correlation between blood gas analysis and chest X-ray lung opacities in young children presenting with acute respiratory symptoms. METHODS: Eighty patients (43 males and 37 females aged 0.5-24 months; mean+/-SD 9.1+/-7.2 months), either with acute wheezing respiratory symptoms and/or with crackles were enrolled in our study. In all children, blood gas analysis and chest X-rays were performed within 12 h following admission to the emergency department. RESULTS: In 55 children (68.75%) chest X-rays demonstrated lung opacities. Subjects with normal X-rays had paO2 and SaO2 higher than subjects with lung opacities (p<0.0001 and p=0.0001, respectively). Children with lung opacities almost always presented paO2<80 mm Hg. Sensitivity and specificity for the presence of lung opacities of paO2<80 mm Hg were 81 and 90%, respectively, while sensitivity and specificity of SaO2<95% were 92 and 40%, respectively. paO2<80 mm Hg in association with SaO2<95% had a positive predictive value for the diagnosis of pneumonia of 90.9%. CONCLUSIONS: Our study suggests that blood gas analysis, particularly paO2, may help in predicting the presence of lung opacities in patients aged less than 2 years. However, chest X-rays may still be needed to define the actual extension of opacities as well as the possible concomitant presence of complications. PMID- 15824529 TI - Utility of rapid on-site evaluation of transbronchial needle aspirates. AB - BACKGROUND: Rapid on-site evaluation has been proposed as a method to improve the yield of transbronchial needle aspiration. OBJECTIVES: This study investigated whether on-site analysis facilitates routine diagnostic bronchoscopy in terms of sampling, yield and cost. METHODS: Patients with lesions accessible for transbronchial needle aspiration on computed tomography were investigated. A cytopathologist screened the needle aspirates on site for the presence of diagnostic material. The bronchoscopic sampling process was adjusted according to the results. In 90 consecutive patients with neoplastic disease (n=70; 78%), non neoplastic disease (n=16; 18%) or undiagnosed lesions (n=4; 4%) we aspirated 162 lung tumours or lymph node sites (mediastinal: 7%; tracheobronchial: 68%; other: 25%). In 90 consecutive patients with neoplastic disease (n=70; 78%), non neoplastic disease (n=16; 18%) or undiagnosed lesions (n=4; 4%) we aspirated 162 lung lesions (paratracheal tumours or lymph nodes: 7%; tracheobronchial lymph nodes: 68%; other: 25%). RESULTS: The diagnostic yield of needle aspiration was 77 and 25% in patients with neoplastic and non-neoplastic lesions, respectively. Sampling could be terminated in 64% of patients after needle aspiration had been performed as the only diagnostic modality, and on-site analysis identified diagnostic material from the first site aspirated in 50% of patients. Only in 2 patients (2%) diagnostic aspirates were not recognized on site. On-site analysis was cost effective due to savings for disposable diagnostic tools, which exceeded the extra expense for the on-site cytology service provided. CONCLUSIONS: Rapid on-site analysis of transbronchial aspirates is a highly useful, accurate and cost-effective addition to routine diagnostic bronchoscopy. PMID- 15824530 TI - Catecholamine clearance from alveolar spaces of rat and human lungs. AB - BACKGROUND: Although aerosolized beta-adrenergic agonists have been used as a therapy for the resolution of pulmonary edema, the mechanisms of catecholamine clearance from the alveolar spaces of the lung are not well known. OBJECTIVE: To determine whether catecholamine clearance from the alveolar spaces is correlated with the fluid transport capacity of the lung. METHODS: Albumin solution containing epinephrine (10(-7)M) or norepinephrine (10(-7)M) was instilled into the alveolar spaces of isolated rat and human lungs. Alveolar fluid clearance rate was estimated by the progressive increase in the albumin concentration over 1 h. Catecholamine clearance rate was estimated by the changes in catecholamine concentration and alveolar fluid volume over 1 h. RESULTS: The norepinephrine clearance rate was faster than the epinephrine clearance rate in the rat and human lungs. In the rat lungs, amiloride (a sodium channel blocker) caused a greater decrease in alveolar fluid clearance and epinephrine clearance rate than propranolol (a nonselective beta-adrenergic antagonist). Although propranolol and phentolamine (an alpha-adrenergic antagonist), and 5-(N-ethyl-N isoprophyl)amiloride (a Na+/H+ antiport blocker) changed neither the alveolar fluid clearance nor the norepinephrine clearance rate, amiloride and benzamil (a sodium channel blocker) decreased both clearance rates. As in the rat lungs, amiloride decreased alveolar fluid and norepinephrine clearance rates in the human lungs. CONCLUSION: These results indicate that the catecholamine clearance rate from the alveolar spaces is correlated with alveolar fluid clearance in rat and human lungs. PMID- 15824531 TI - Yellow nail syndrome. PMID- 15824532 TI - Cheyne-Stokes respiration in congestive heart failure: continuous positive airway pressure of 5-8 cm H2O for 1 year in five cases. AB - Five adult patients with congestive heart failure (CHF) due to dilated cardiomyopathy complicated by Cheyne-Stokes respiration/central sleep apnea (CSR/CSA) were treated with continuous positive airway pressure (CPAP) with an initial pressure of 5 cm H2O. Four patients were followed up for 12 months with CPAP of 5 cm H2O. The rest, a 93-year-old patient, was followed up for 30 months, and the CPAP was reset at 8 cm H2O due to worsened cardiac function after 6 months and it was reset at 6 cm H2O due to dryness of the nose after 23 months. For all the patients with nightly CPAP use for 6.0+/-1.4 h per day for a year, frequency of CSR/CSA was significantly reduced after 3 and 12 months with CPAP (p<0.05). Moreover, their symptoms, cardiac function and sleep quality were significantly improved after 3 months (p<0.05), and were maintained above the pre CPAP levels after 12 months, except for the oldest patient whose cardiac function tended to deteriorate. The results suggest that CSR/CSA in CHF can be treated with CPAP set at a lower pressure than the conventional method, and that CPAP at 5-8 cm H2O is often effective in eliminating CSR/CSA, improving sleep quality, and presumably maintaining cardiac function. PMID- 15824533 TI - Immunosuppressant-responsive idiopathic lymphocytic pleuritis. AB - Lymphocyte-dominant pleural inflammation is seen in tuberculous, malignant and immune-mediated pleural disease. We describe two male patients who presented with chest pain and dyspnoea, in whom radiological and histological investigations showed idiopathic lymphocytic pleuritis that responded to immunosuppressive therapy with corticosteroids and/or azathioprine. Corticosteroid-responsive pleuritis has been described in association with many conditions, but little is known of idiopathic lymphocytic pleuritis. The authors suggest that patients with idiopathic pleural thickening undergo early thoracoscopic pleural biopsy, and that immunosuppressant therapy be considered for patients with idiopathic lymphocytic pleuritis. PMID- 15824534 TI - Acute respiratory failure with an abnormal chest radiograph. PMID- 15824535 TI - Psychological implications of respiratory health and disease. AB - The possibility that a subject's psychological status may influence respiratory sensations and that chronic respiratory disease may have psychological consequences has sparked great interest among clinicians and researchers. This paper reviews the existing research on the association between respiratory symptom perception and the psychological status and between chronic respiratory diseases, such as asthma and chronic obstructive pulmonary disease, and psychological disturbances. Moreover, it focuses on the role of stressful events in determining asthma exacerbations. The recent literature suggests that in patients with chronic respiratory diseases, the evaluation of breathlessness perception, psychological disturbances and the recording of any stressful events should be considered as relevant as the physical and functional assessment of respiration. PMID- 15824536 TI - Can reduced smoking be a way for smokers not interested in quitting to actually quit? AB - The predominating way to stop smoking is to do it abruptly. At every given time, the large majority of smokers are not motivated or willing to try and give up. Some smokers are entirely happy with their smoking, a larger group would like to smoke less and a third group wants to quit. With the abrupt quitting message we are only addressing those wanting to quit. Maybe not even all, since some of them may have tried many times already and learned that they cannot quit abruptly. They may have given up on giving up. Some interesting results are given in recent studies that have recruited smokers not motivated to quit but interested in reducing their smoking. From nine randomized placebo-controlled trials where smokers were given behavioural support and pharmacological assistance, motivation to quit seemed to have increased, and in each trial, a proportion of these unmotivated smokers gave up smoking. It is suggested that for smokers unable or not interested in giving up abruptly, a softer and more gradual approach should be considered. Such an approach may bring new smokers into treatment, produce more people wanting to quit and improve public health. PMID- 15824537 TI - Outbreak of Bronchiolitis obliterans associated with consumption of Sauropus androgynus in Japan--alert of food-associated pulmonary disorders from Japan. PMID- 15824538 TI - Miguel Hernan, winner of the Kenneth Rothman epidemiology prize, 2005. PMID- 15824539 TI - Our policy on conflict of interest: open for discussion. PMID- 15824540 TI - HIV and mortality of mothers and children: evidence from cohort studies in Uganda, Tanzania, and Malawi. AB - BACKGROUND: The steady decline in child mortality observed in most African countries through the 1960s, 1970s, and 1980s has stalled in many countries in the 1990s because of the AIDS epidemic. However, the census and household survey data that generally are used to produce estimates of child mortality do not permit precise measures of the adverse effect of HIV on child mortality. METHODS: To calculate excess risks of child mortality as the result of maternal HIV status, we used pooled data from 3 longitudinal community-based studies that classified births by the mother's HIV status. We also estimated excess risks of child death caused by increased mortality among mothers. The joint effects of maternal HIV status and maternal survival were quantified using multivariate techniques in a survival analysis. RESULTS: Our analysis shows that the excess risk of death associated with having an HIV-positive mother is 2.9 (95% confidence interval = 2.3-3.6), and this effect lasts throughout childhood. The excess risk associated with a maternal death is 3.9 (2.8-5.5) in the 2-year period centered on the mother's death, with children of both infected and uninfected mothers experiencing higher mortality risks at this time. CONCLUSION: HIV impacts on child mortality directly through transmission of the virus to newborns by infected mothers and indirectly through higher child mortality rates associated with a maternal death. PMID- 15824541 TI - Thrombophilic polymorphisms and intrauterine growth restriction. AB - BACKGROUND: The relationship between thrombophilic polymorphisms and intrauterine growth restriction remains unclear. Whereas a subset of these polymorphisms have received some attention, others have not. METHODS: We conducted a case-control study of 493 cases of intrauterine growth restriction (birthweight less than the 10th percentile for gestational age and sex) and 472 controls (greater than the 10th percentile). We also conducted a family study including approximately 250 case trios (affected newborn, mother, and father) for each studied gene. Plasminogen activator inhibitor-1(PAI-1) 4G/5G and Factor XIII Val134Leu variants were compared between maternal and newborn cases and controls. Relative risks for newborn and maternal PAI-1 and Factor XIII genotypes in case trios were estimated using a log-linear model. Transmission of MTHFR C677T and 1298C haplotypes was analyzed in case trios, and the estimated haplotype distribution compared between cases and controls. Finally, we explored pairwise gene-gene interactions between all measured polymorphisms, including Factor V Leiden G1691A and Prothrombin G20210A. RESULTS: PAI-1 and Factor XIII polymorphisms were not associated with an increased risk of intrauterine growth restriction in the case-control analysis, and the case-parent analysis showed no newborn or maternal excess genotype relative risk. No excess transmission of the MTHFR haplotypes was observed in case newborns, and the distribution of MTHFR haplotypes was similar between cases and controls. Some results were suggestive of interactions between genes. CONCLUSIONS: Overall, there seems to be little or no indication that thrombophilic genes, at least individually, have an effect on intrauterine growth restriction. PMID- 15824542 TI - Maternal smoking and birth weight: interaction with parity and mother's own in utero exposure to smoking. AB - BACKGROUND: Few studies have reported interactions between maternal smoking and other maternal characteristics and exposures. We examined maternal smoking in a cohort study for which data from 3 generations were available to examine maternal characteristics and exposures from a life-course perspective. METHODS: We had data from 3 generations: women enrolled in the U.S. Collaborative Perinatal Project (CPP) between 1959 and 1965 at the Baltimore site (G1); daughters (G2) of those G1 mothers who were followed to ages 27-33 years in the Pathways to Adulthood study; and children (G3) born to the G2 women who provided pregnancy and birth information during the Pathways study. These data allowed examination of exposures that occurred to the mother during her childhood and in utero. RESULTS: We found evidence of a 3-way interaction effect on birth weight for maternal smoking in pregnancy, maternal exposure to smoking in utero (grandmaternal smoking), and maternal parity. Maternal smoking reduced birth weight in 3 of the subgroups, with the size of the effect on birth weight moderated by parity and the mother's own in utero exposure to smoking. CONCLUSIONS: A mother's prenatal exposure to smoke may affect the birth weight of her offspring. This effect would be consistent with both the accumulation-of-risk and the fetal-programming hypotheses. PMID- 15824543 TI - Assessing maternal genetic associations: a comparison of the log-linear approach to case-parent triad data and a case-control approach. AB - BACKGROUND: In utero exposures, including maternal phenotypes, are potential risk factors for both early-onset and adult-onset diseases. Two alternative study designs use maternal genotypes at polymorphic loci as biomarkers of an offspring's in utero exposure: (1) a traditional case-control study with logistic regression analysis, in which cases, controls, and mothers of both types of subjects are genotyped; and (2) a case-parent triad study with log-linear analysis, in which cases and both parents are genotyped. METHODS: We used computer simulations to compare the operating characteristics of the log-linear approach to case-parent triad data and the case-control approach for assessing relative risks (RRs) associated with maternal genotypes. RESULTS: For high-risk allele frequencies (chromosomal prevalence; f) between 0.20 and 0.75, both methods allowed for valid, unbiased estimates of maternal RRs. The case-parent triad approach, however, had 43% greater power, on average, than the case-control approach with an equal number of genotypes, and 13% greater power with an equal number of cases. For example, under dominant inheritance, to detect 2-fold maternal RRs with 200 (or 150) cases when allele prevalence is between 0.15 and 0.40, the case-parent triad and equal-genotype case-control designs had, on average, 87% and 62% power, respectively. As f approached 0 or 1, the power of both methods decreased sharply. DISCUSSION: The greater efficiency of case-parent triads may be due to the inclusion of paternal genotype information, which allows for independent tests of disease association with maternal or offspring genotypes. These results highlight one potential advantage of case-parent triad data in assessing maternal genetics as risk factors for offspring disease. We discuss these findings and other considerations between the 2 methodological approaches. PMID- 15824544 TI - Does prenatal sonography affect intellectual performance? AB - BACKGROUND: Little is known about potential risks of ultrasound during pregnancy. The developing brain is susceptible to environmental influences, and prenatal ultrasound has been reported to affect cognitive abilities in animals. METHODS: To assess a possible association between prenatal ultrasound and intellectual performance, we studied men born in Sweden from 1973 to 1978 who enrolled for military service from 1991 to 1996. Intellectual scores (mean +/- standard deviation = 5 +/- 2) were measured by a test battery at enrollment, and subnormal performance was defined as a score of 3 or less. We estimated differences in mean intellectual score between ultrasound-exposed and ultrasound-unexposed using linear regression, and we calculated relative risks of subnormal performance using logistic regression analysis. RESULTS: There were 7999 eligible men born in a hospital (Malmo) that included ultrasound scanning in standard antenatal care (exposed) and 197,829 men born in hospitals without ultrasound scanning programs (unexposed). We found lower intellectual performance scores (mean difference = 0.16; 95% confidence interval = -0.21 to -0.11) and an increased risk of subnormal performance (odds ratio = 1.28; CI = 1.18 to 1.38) among ultrasound exposed compared with unexposed. However, men born in Malmo before scanning was introduced also had lower scores, and the decrease in test scores after the introduction of ultrasound was small. Moreover, we found no differences in intellectual performance within pairs of brothers as a result of ultrasound exposure. CONCLUSIONS: This study failed to demonstrate a clear association between ultrasound scanning and intellectual performance. PMID- 15824545 TI - Parent's age and the risk of oral clefts. AB - BACKGROUND: Some malformations are clearly associated with older maternal age, but the effect of older age of the father is less certain. The aim of this study is to determine the degree to which maternal age and paternal age independently influence the risk of having a child with oral clefts. METHODS: Among the 1,489,014 live births in Denmark during 1973-1996, there were 1920 children with nonsyndromic cleft lip with or without cleft palate and 956 children with nonsyndromic cleft palate. We used logistic regression to assess the impact of parental age on the occurrence of cleft lip with or without cleft palate and cleft palate. Interaction between mother's and father's age was included in the analysis. RESULTS: Separate analyses of mother's and father's age showed that older age was associated with increased risk of both cleft lip with or without cleft palate and cleft palate only. In a joint analysis, both maternal and paternal ages were associated with the risk of cleft lip with or without cleft palate, but the contribution of each was dependent on the age of the other parent. In the analysis of cleft palate only, the effect of maternal age disappeared, leaving only paternal age as a risk factor. CONCLUSION: Both high maternal age and high paternal age were associated with cleft lip with or without cleft palate. Higher paternal age but not maternal age increased the risk of cleft palate only. PMID- 15824546 TI - Eye anomalies: seasonal variation and maternal viral infections. AB - INTRODUCTION: Eye anomalies, including anophthalmos and microphthalmos (absent eye and small eye), are common features in congenital infection syndromes such as rubella. Influenza, Coxsackie virus, and Parvovirus are less well-established teratogens. We report an ecologic analysis of anophthalmia/microphthalmia prevalence in England in relation to temporal variation in these infections using routine infection data. METHODS: The national (England) anophthalmos/microphthalmos register holds details of 406 babies (excluding chromosomal anomalies) born 1988-1994. Weekly infection data were obtained from the U.K. Public Health Laboratory Service and Royal College of General Practitioners Weekly Returns Service. We assigned cases and births an estimated exposure based on the population infection counts in the gestational weeks known to be critical for ocular development. We calculated the rate ratio of anophthalmos/microphthalmos prevalence for infection in the 90th percentile compared with the 10th percentile by Poisson regression. RESULTS: There was a positive association between severe anophthalmos/microphthalmos prevalence and Parvovirus B19 (rate ratio = 1.26; 95% confidence interval = 1.00-1.58) and for influenza (1.41; 1.08-1.84). Coxsackie virus showed a negative association with any anophthalmos/microphthalmos (0.75; 0.58-0.98) and with severe cases (0.66; 0.46-0.95). CONCLUSIONS: This analysis provides suggestive evidence for a teratogenic role of influenza and Parvovirus in anophthalmos/microphthalmos. The role of maternal infections in congenital anomaly etiology is worthy of further investigation. PMID- 15824547 TI - Preterm delivery in Boston before and after September 11th, 2001. AB - BACKGROUND: We hypothesized that stress induced by the terrorist attacks of September 11, 2001 might shorten pregnancy. To test this hypothesis, we compared gestational duration and risk of preterm delivery among women who were pregnant on September 11 with women who had delivered before that date. STUDY DESIGN: We conducted a matched cohort study among pregnant women enrolled in the Boston based cohort study Project Viva between 1999 and 2001. Each of 606 participants, pregnant on September 11, 2001, was matched to 1 or 2 participants who delivered before that date. RESULTS: Compared with women who delivered before September 11th, women who were pregnant on September 11th had mean gestation length that was 0.13 weeks longer (95% confidence interval = -0.05 to 0.30) and an odds ratio for preterm delivery before 37 weeks' gestation of 0.60 (0.36 to 0.98). Only women exposed in the first trimester had longer gestation. CONCLUSIONS: Contrary to expectation, Boston-area women who were pregnant on September 11th had a lower risk of preterm delivery than women who delivered before that date. Although the interpretation of this finding is difficult, it is clear that the acute psychologic stress documented nationwide after the terrorist attacks did not increase the risk of preterm delivery in this population at some distance from the attacks. PMID- 15824548 TI - Health consequences of breast-feeding: doctors' visits and hospitalizations during the first 18 months of life in Hong Kong Chinese infants. AB - BACKGROUND: The evidence on whether breast-feeding reduces health services use in nonwhite infants is scant. We examined the effects of breast-feeding on health services utilization in Hong Kong Chinese infants. METHODS: We followed a population-based cohort of 8327 infants born in 1997 for 18 months. The main outcome measures were higher (above the sample mean) utilization of outpatient visits and hospitalizations for jaundice, gastrointestinal or respiratory/febrile illnesses, and all illnesses. RESULTS: Breast-fed infants had fewer illness related doctor visits overall through the first 18 months of life. Results were strongest for infants breast fed exclusively for 2 to 3 months (odds ratio [OR] for higher utilization = 0.78; 95% confidence interval [CI] = 0.62-0.99) and for 4 or more months (0.65; 0.53-0.81). However, breast-fed infants were more likely to receive outpatient care for jaundice, particularly in the first 3 months of life (ORs ranging from 2.5 to 8.4). Any breast-feeding was also associated with more jaundice-related hospital admissions, the effects of which were most acute in the first 3 months of life. Compared with exclusively formula-fed infants, the OR (CI) for mixed breast- and formula-fed was 2.4 (1.7-3.5); for exclusive breast feeding up to 1 month, 4.5 (2.7-7.6); for exclusive breast-feeding 2 to 3 months, 3.2 (1.8-5.7); and for exclusive breast-feeding 4 or more months, 3.4 (2.0-5.7). CONCLUSIONS: Breast-feeding in Hong Kong Chinese infants reduces doctor visits overall, but increases both outpatient visits and hospitalizations for jaundice. PMID- 15824549 TI - Active surveillance of vaccine safety: a system to detect early signs of adverse events. AB - BACKGROUND: There currently are no population-based systems in the United States to rapidly detect adverse events after newly introduced vaccines. To evaluate the feasibility of developing such systems, we used 5 years of data from 4 health maintenance organizations within the Centers for Disease Control and Prevention (CDC) Vaccine Safety Datalink. METHODS: Within every year, each week's vaccinated children were followed for 4 weeks, and rates of adverse events were compared with rates among children of similar ages before the introduction of the new vaccine. We assessed risks for intussusception after rotavirus vaccination and risks for fever, seizures, and other neurologic adverse events after the change from whole cell diphtheria-tetanus-pertussis (DTPw) to acellular DTP vaccine (DTPa). We used sequential probability ratio testing, adjusted for age, sex, calendar time, season, and HMO, and with a stopping value based on the probability of an adverse event under the null hypothesis and under a preset alternative hypothesis. RESULTS: We detected an increase in intussusception after 2589 vaccine doses of rotavirus vaccine, about the same time initial reports of intussusception were made to the Vaccine Adverse Events Reporting System. Decreases in risk for fever, seizures, and other abnormal neurologic events became detectable within 12 weeks, 42 weeks, and 18 months, respectively, after the change from DTPw to DTPa. CONCLUSIONS: We conclude that it is feasible to develop systems for rapid and routine population-based assessments of new vaccine safety. PMID- 15824550 TI - Breast cancer incidence in women prenatally exposed to maternal cigarette smoke. AB - BACKGROUND: Clinical studies show that maternal cigarette smoking reduces pregnancy estrogen levels. Women prenatally exposed to maternal cigarette smoke may, therefore, have a lower breast cancer risk because the fetal mammary gland's exposure to maternal estrogen is decreased. Associations between prenatal maternal cigarette smoke exposure and breast cancer, however, have not been observed in previous case-control studies that relied on exposure assessment after the onset of cancer. At the start of this study, cigarette smoking history was obtained directly from the mother. METHODS: The National Cooperative DES Adenosis project was a follow-up study of health outcomes in women prenatally exposed to diethylstilbestrol (DES). At the start of the study, women's mothers provided information about cigarette smoking habits during the time they were pregnant with the study participant. In the current study, the breast cancer rates are compared among 4031 women who were or were not prenatally exposed to maternal cigarette smoke. The resultant relative rate (RR) is adjusted for potential confounding by other breast cancer risk factors using Poisson regression modeling. RESULTS: Fetal exposure to maternal cigarette smoke appeared to be inversely associated with breast cancer incidence (RR = 0.49; 95% confidence interval [CI] = 0.24-1.03). The inverse association was more apparent among women whose mothers smoked 15 cigarettes or fewer per day than among daughters of heavier smokers. There were, however, too few cases to precisely estimate a possible dose-response relationship. CONCLUSION: These results support the hypothesis that in utero exposure to maternal cigarette smoke reduces breast cancer incidence. PMID- 15824551 TI - Influence of body size and body fat distribution on risk of uterine leiomyomata in U.S. black women. AB - BACKGROUND: Uterine leiomyomata are a major source of morbidity in black women. We prospectively investigated the risk of self-reported uterine leiomyomata in relation to body mass index (BMI), weight change, height, waist and hip circumferences, and waist-to-hip ratio in a large cohort of U.S black women. METHODS: Data were derived from the Black Women's Health Study, a U.S. prospective cohort study of black women who complete biannual mailed health questionnaires. From 1997 through 2001, we followed 21,506 premenopausal women with intact uteri and no prior diagnosis of uterine leiomyomata. Cox regression models were used to estimate incidence rate ratios (IRRs) and 95% confidence intervals (CIs). RESULTS: After 70,345 person-years of follow up, 2146 new cases of uterine leiomyomata confirmed by ultrasound (n = 1885) or hysterectomy (n = 261) were self-reported. Compared with the thinnest women (BMI <20.0 kg/m), the multivariate IRRs for women with BMIs of 20.0-22.4, 22.5-24.9, 25.0-27.4, 27.5 29.9, 30.0-32.4, and 32.5+ kg/m were 1.34 (95% CI = 1.02-1.75), 1.39 (1.07-1.81), 1.45 (1.12-1.89), 1.47 (1.11-1.93), 1.36 (1.02-1.80), and 1.21 (0.93-1.58), respectively. IRRs were larger among parous women. Weight gain since age 18 was positively associated with risk, but only among parous women. No other anthropometric measures were associated with risk. CONCLUSIONS: BMI and weight gain exhibited a complex relation with risk of uterine leiomyomata in the Black Women's Health Study. The BMI association was inverse J-shaped and findings were stronger in parous women. Weight gain was positively associated with risk among parous women only. PMID- 15824552 TI - The cross-sectional cohort study: an underutilized design. AB - Epidemiologists frequently study the experience of a population over time to estimate the association between exposure and outcome. These studies generally use case-control, prospective cohort, or retrospective cohort designs. The "cross sectional cohort study," as it is termed here, represents an alternative to these standard methods. With this design, an investigator samples a source population cross-sectionally and then retrospectively assesses subjects' histories of exposures and outcomes over a specified time period. Certain threats to validity, such as nonignorable exiting and measurement error from retrospective assessments, must be considered carefully when using this design. However, in some situations, the cross-sectional cohort design may offer advantages over traditional designs-especially in studies in which there is a long interval between exposures and outcomes, in which exposures and outcomes can be accurately assessed retrospectively, and in which the outcome does not accelerate any exiting from the population. Such situations often arise when studying chronic or episodic conditions with low mortality, such as psychiatric disorders. PMID- 15824553 TI - Respiratory medications and the risk of cardiac arrhythmias. AB - BACKGROUND: Medications used to treat respiratory diseases include beta adrenoceptors, antimuscarinics, inhaled and oral corticosteroids, and theophyllines. Most of these drugs have been associated indirectly with cardiac rhythm disorders, but epidemiologic evidence is limited. METHODS: To evaluate the association between respiratory drugs and the occurrence of rhythm disorders among patients with asthma and those with chronic obstructive pulmonary disease, we conducted a case-control study nested in a population-based cohort of individuals 10-79 years of age and registered in the U.K. General Practice Research Database after 1 January 1994. The analysis included 710 confirmed cases and 5000 controls frequency-matched to cases by age (interval of 1 year) and sex. RESULTS: No increased risk of arrhythmias overall was found among users of inhaled steroids (relative risk = 1.0; 95% confidence interval = 0.8-1.3). Short term use of theophylline was weakly associated with arrhythmia (1.8; 1.0-3.3). An increased risk was found among users of oral steroids, and the relative risk was greater at the beginning of therapy (2.6; 2.0-3.5). The risk of atrial fibrillation was increased, especially for short-term use of oral steroids (2.7; 1.9-3.8), and a weak association was seen for theophyllines, especially short term use (1.8; 0.9-3.7). Supraventricular tachycardia was associated with long term use of oral steroids (2.1; 0.8-5.7), long-term use of antimuscarinics (1.7; 0.7-4.1), and short-term use of theophylline (4.0; 0.9-18.1). Ventricular arrhythmias were associated with oral steroids (3.2; 0.8-13.3) and beta adrenoceptors (7.1; 0.8-65.9). CONCLUSIONS: Oral steroids and theophylline were the therapeutic groups associated with risk of developing atrial fibrillation, especially with new courses of therapy. Results from this study also are consistent with certain suspected dysrhythmic effects of theophyllines, with supraventricular tachycardia associated with antimuscarinics, and with ventricular arrhythmias associated with beta-adrenoceptors. PMID- 15824554 TI - Screening by prostate-specific antigen and digital rectal examination in relation to prostate cancer mortality: a case-control study. AB - BACKGROUND: The potential role of prostate cancer screening in reducing mortality is uncertain. To examine whether screening with the prostate-specific antigen (PSA) test or digital rectal examination is associated with reduced prostate cancer mortality, we conducted a population-based case-control study in 4 health maintenance organizations. METHODS: Cases were 769 health plan members who died because of prostate adenocarcinoma during the years 1997-2001. We randomly selected 929 controls from the health plan membership and matched them to cases on health plan, age, race, and membership history. Medical records were used to document all screening tests in the 10 years before and including the date on which prostate cancer was first suspected. RESULTS: Among white participants, 62% of cases and 69% of controls had a least 1 screening PSA test or digital rectal examination (odds ratio = 0.73; 95% confidence interval = 0.55-0.97). The corresponding proportions for blacks were 59% and 61% (1.0; 0.59-1.4). Most screening tests were digital rectal examinations; therefore, in the subgroup with no history of PSA screening, the association between digital rectal screening and prostate cancer mortality was similar to the overall association (0.65 [0.48 0.88] among whites; 0.86 [0.53-1.4] among blacks). Very few men received screening PSA without screening digital rectal examination (6% of cases and 7% of controls among whites). CONCLUSIONS: Digital rectal screening was associated with a reduced risk of death due to prostate cancer in our population. Because of several data limitations, this study could not accurately estimate the effect of PSA screening separate from digital rectal examination. PMID- 15824555 TI - Indoor heating sources and respiratory symptoms in nonsmoking women. AB - BACKGROUND: Secondary heating appliances are important indoor sources of air pollution, including particulate matter, nitrogen dioxide (NO2), and sulfur dioxide (SO2). We hypothesized that the use of secondary heating sources increases respiratory symptoms in women living in nonsmoking households and specifically that concentrations of SO2 and NO2 emitted from heating sources are associated with respiratory symptoms. METHODS: Mothers who delivered babies at 12 hospitals in Connecticut and Virginia (1993-1996) were enrolled. There were 888 women who contributed symptom and exposure information during the winter heating season (15 October to 15 April), for a total of 9783 reporting periods (median = 12 reporting periods per woman, interquartile range 11-12). Adjusted rate ratios (RRs) of effects of source use and measured concentrations on rate of days with symptoms were obtained using generalized estimating equations for a log-linear Poisson model, controlling age, education, race, history of allergies, number of children, dwelling type, and residence state. RESULTS: In adjusted models, each hour-per-day increase in kerosene heater use is associated with an increase in wheezing (RR = 1.06; 95% confidence interval (CI) = 1.01-1.11). Each hour of fireplace use is associated with increased cough (1.05; 1.01-1.09), sore throat (1.04; 1.00-1.08), and marginally with chest tightness (1.05; 0.99-1.12). Each 10 ppb increase in SO2 (a proxy for sulfate aerosol) is associated with increased wheezing (1.57; 1.10-2.26) and chest tightness (1.32; 1.01-1.71). CONCLUSIONS: Emissions from fireplaces, gas space heaters, and kerosene heaters may contribute to respiratory symptoms in a population of nonsmoking women. PMID- 15824556 TI - Ambient gas concentrations and personal particulate matter exposures: implications for studying the health effects of particles. AB - BACKGROUND: Data from a previous study conducted in Baltimore, MD, showed that ambient fine particulate matter less than 2.5 mum in diameter (PM2.5) concentrations were strongly correlated with corresponding personal PM2.5 exposures, whereas ambient O3, NO2, and SO2 concentrations were weakly correlated with their personal exposures to these gases. In contrast, many of the ambient gas concentrations were reasonable surrogates of personal PM2.5 exposures. METHODS: Personal multipollutant exposures and corresponding ambient air pollution concentrations were measured for 43 subjects living in Boston, MA. The cohort consisted of 20 healthy senior citizens and 23 schoolchildren. Simultaneous 24-hour integrated PM2.5, O3, NO2, and SO2 personal exposures and ambient concentrations were measured. All PM2.5 samples were also analyzed for SO4 (sulfate). We analyzed personal exposure and ambient concentration data using correlation and mixed model regression analyses to examine relationships among (1) ambient PM2.5 concentrations and corresponding ambient gas concentrations; (2) ambient PM2.5 and gas concentrations and their respective personal exposures; (3) ambient gas concentrations and corresponding personal PM2.5 exposures; and (4) personal PM2.5 exposures and corresponding personal gas exposures. RESULTS: We found substantial correlations between ambient PM2.5 concentrations and corresponding personal exposures over the course of time. Additionally, our results support the earlier finding that summertime gaseous pollutant concentrations may be better surrogates of personal PM2.5 exposures (especially personal exposures to PM2.5 of ambient origin) than they are surrogates of personal exposures to the gases themselves. CONCLUSIONS: Particle health effects studies that include both ambient PM2.5 and gaseous concentrations as independent variables must be analyzed carefully and interpreted cautiously, since both parameters may be serving as surrogates for PM2.5 exposures. PMID- 15824557 TI - Exposure to ambient and nonambient components of particulate matter: a comparison of health effects. AB - BACKGROUND: Numerous epidemiologic studies report associations between outdoor concentrations of particles and adverse health effects. Because personal exposure to particles is frequently dominated by exposure to nonambient particles (those originating from indoor sources), we present an approach to evaluate the relative impacts of ambient and nonambient exposures. METHODS: We developed separate estimates of exposures to ambient and nonambient particles of different size ranges (PM2.5, PM10-2.5 and PM10) based on time-activity data and the use of particle sulfate measurements as a tracer for indoor infiltration of ambient particles. To illustrate the application of these estimates, associations between cardiopulmonary health outcomes and the estimated exposures were compared with associations computed using measurements of personal exposures and outdoor concentrations for a repeated-measures panel study of 16 patients with chronic obstructive pulmonary disease conducted in the summer of 1998 in Vancouver. RESULTS: Total personal fine particle exposures were dominated by exposures to nonambient particles, which were not correlated with ambient fine particle exposures or ambient concentrations. Although total and nonambient particle exposures were not associated with any of the health outcomes, ambient exposures (and to a lesser extent ambient concentrations) were associated with decreased lung function, decreased systolic blood pressure, increased heart rate, and increased supraventricular ectopic heartbeats. Measures of heart rate variability showed less consistent relationships among the various exposure metrics. CONCLUSIONS: These results demonstrate the usefulness of separating total personal particle exposures into their ambient and nonambient components. The results support previous epidemiologic findings using ambient concentrations by demonstrating an association between health outcomes and ambient (outdoor origin) particle exposures but not with nonambient (indoor origin) particle exposures. PMID- 15824559 TI - Science policy and politics: will the recent past preface the future? PMID- 15824560 TI - A conversation with Lester Breslow. PMID- 15824561 TI - Mobile phone use and acoustic neuromas. PMID- 15824562 TI - Mobile phone use and acoustic neuromas. PMID- 15824563 TI - Mobile phone use and acoustic neuromas. PMID- 15824564 TI - Mobile phone use and acoustic neuromas. PMID- 15824565 TI - Mobile phone use and acoustic neuromas. PMID- 15824567 TI - "Black-box" epidemiology. PMID- 15824568 TI - "Black-box" epidemiology. PMID- 15824569 TI - Folic acid and placental pathology. PMID- 15824571 TI - Birth weight and intima-media thickness. PMID- 15824572 TI - Air pollution and breast cancer. PMID- 15824574 TI - Hepatitis C among intravenous drug users. PMID- 15824576 TI - Deterministic vs. probabilistic analyses to identify sensitive parameters in dose assessment using RESRAD. AB - The dose assessments for sites containing residual radioactivity usually involve the use of computer models that employ input parameters describing the physical conditions of the contaminated and surrounding media and the living and consumption patterns of the receptors in analyzing potential doses to the receptors. The precision of the dose results depends on the precision of the input parameter values. The identification of sensitive parameters that have great influence on the dose results would help set priorities in research and information gathering for parameter values so that a more precise dose assessment can be conducted. Two methods of identifying site-specific sensitive parameters, deterministic and probabilistic, were compared by applying them to the RESRAD computer code for analyzing radiation exposure for a residential farmer scenario. The deterministic method has difficulty in evaluating the effect of simultaneous changes in a large number of input parameters on the model output results. The probabilistic method easily identified the most sensitive parameters, but the sensitivity measure of other parameters was obscured. The choice of sensitivity analysis method would depend on the availability of site-specific data. Generally speaking, the deterministic method would identify the same set of sensitive parameters as the probabilistic method when 1) the baseline values used in the deterministic method were selected near the mean or median value of each parameter and 2) the selected range of parameter values used in the deterministic method was wide enough to cover the 5th to 95th percentile values from the distribution of that parameter. PMID- 15824577 TI - Design of an inexpensive, flexible, shielded cave for environmental radioactivity measurements. AB - A graded-absorber radiation shield (cave) was designed with low cost and flexibility to lower background for counting environmental and other samples containing small amounts of radioactivity. PMID- 15824578 TI - The University of Michigan Student Health Physics Society's Radiation and Health Physics World Wide Web Site. AB - The University of Michigan Student Health Physics Society's (UMSHPS) Radiation and Health Physics World Wide Web Site is an informative database of radiation and health physics related topics. With over 1,000 visitors each day, the UMSHPS web site provides professionals and the general public with a valuable resource for information and research. Users of this site can either search for information by topic or submit questions directly to the qualified members the national Health Physics Society. During the past year, progress has been made in replacing the site's older, less versatile framework with new search engines and refined submittal forms, as well as a "Frequently Asked Questions" section. Within the database, references will include brief summaries of the site's available information and target audience. Although these changes have been beneficial for the site, the UMSHPS continuously seeks professional opinions and ideas to further the services that this online resource can provide to the profession and to the general public. PMID- 15824581 TI - Hands and forearms, feet and ankles. PMID- 15824582 TI - ORS interview with James Graham, RSO and Associate Director of Environmental Health Services at Colorado State University. PMID- 15824583 TI - A procedure for doing radiological surveys of shipping containers. PMID- 15824585 TI - Risk-informed, performance-based inspections at medical facilities. AB - During the past couple of years, radiation safety professionals have observed a significant change with regard to the inspection philosophy of regulators. The NRC and many Agreement State agencies have implemented a performance-based, risk informed approach for inspecting medical Radiation Safety Programs. This new, less prescriptive approach originates from the necessity to produce safety benefits commensurate with their cost to the industry and still maintain health and safety performance. While compliance with regulatory requirements is important, regulatory agencies have been focusing on areas that provide greater safety benefit, such as protecting the radiation worker, members of the general public and the environment. This paper discusses simple and practical measures that may assist licensees in preparing for performance-based, risk informed inspections. PMID- 15824586 TI - An algorithm for pre- and post-irradiation fade in the Thermo 8802 LiF:Mg,Ti thermoluminescent dosimeter. AB - The diminished response of thermoluminescent phosphors over time is a well documented challenge to thermoluminescent dosimetry. Wide ranges in fading rates for various phosphor types have been reported, making it necessary for many external dosimetry programs to perform individual studies on thermoluminescent fade. Sandia National Laboratories currently uses the Thermo 8802 LiF:Mg,Ti thermoluminescent dosimeter (TLD) in its personnel external dosimetry program. Doses received in the field are calculated by applying a fade algorithm published by the manufacturer to TLD readings. Since the algorithm was established by characterizing the diminished response of a TLD similar to the 8802, Sandia chose to model its fade study after the analysis done by Thermo. As a result, the parameters of each experiment were comparable, and data from the two studies were compared to determine whether or not the current algorithm should be modified specifically for use at Sandia. Cards were irradiated using an internal Sr/Y source, and pre- and post-irradiation fading rates were monitored over a period of 18 wk. While significant fading was demonstrated, results closely matched those found in the original Thermo study. PMID- 15824587 TI - Medical radiography at sea. AB - The medical use of x-rays on cruise ships presents some interesting challenges to those used to radiation protection on land. However, the same radiation protection principles apply in both situations. The protection of staff and public apply wherever x-ray machines are used. Whether medical staff, patients, other crew or passengers, all need to be considered with respect to restriction of exposure. An important part of patient dose restriction is dose assessment. A procedure for doing this is presented in this article. The physical assessment of an x-ray facility has two aspects: assessment of shielding afforded by the room structure and assessment of the x-ray equipment itself. Assessment of ancillary equipment is also appropriate; for example, the film processor can have a direct impact on doses to patients. Badly processed films might necessitate repeat exposures. Underdeveloped films might also mislead operators into suspecting that the film is underexposed--the apparent solution is an increase in output causing an unacceptable increase in patient dose. This article also discusses some aspects of the design and assessment of x-ray facilities and the adaptations necessary to achieve a reasonable level of protection in an unusual environment. PMID- 15824588 TI - Test of radiation detectors used in homeland security applications. AB - This work was performed as part of the National Institute of Standards and Technology (NIST) program to support the development of the new American National Standards Institute (ANSI) standards N42.32-2003 and N42.33-2003 for hand-held detectors, and personal electronic dosimeters, as well as to support the Office of Law Enforcement Standards (OLES) and the Department of Homeland Security (DHS) in testing these types of detectors for their use by first responders. These instruments are required to operate over a photon energy range of 60 keV to 1.33 MeV and over a wide range of air-kerma rates. The performance and response of various radiation detectors, purchased by the NIST, was recorded when placed in 60Co, 137Cs, and x-ray beams at different air-kerma rates. The measurements described in this report were performed at the NIST x-ray and gamma-ray radiation calibration facilities. The instruments' response (exposure or dose rate readings) shows strong energy dependence but almost no dependence to different air-kerma rates. The data here reported provide a benchmark in support of current protocols that are being developed for radiation detection instrumentation used in homeland security applications. A future plan is to test these devices, plus other commercially available detectors, against ANSI standards N42.32-2003 and N42.33-2003. PMID- 15824589 TI - High level waste tank closure project: ALARA applications at the Idaho National Engineering and Environmental Laboratory. AB - Bechtel BWXT Idaho, Maintenance and Operating Contractor for the Department of Energy at the Idaho National Engineering and Environmental Laboratory, has emptied, cleaned, and sampled six of the eleven 1.135 x 10(6) L high level waste underground storage tanks at the Idaho Nuclear Technology and Engineering Center, well ahead of the State of Idaho Consent Order cleaning schedule. Cleaning of a seventh tank is expected to be complete by the end of calendar year 2004. The tanks, with associated vaults, valve boxes, and distribution systems, are being closed to meet Resource Conservation and Recovery Act regulations and Department of Energy orders. The use of remotely operated equipment placed in the tanks through existing tank riser access points, sampling methods and application of as low-as-reasonably-achievable (ALARA) principles have proven effective in keeping personnel dose low during equipment removal, tank, vault, and valve box cleaning, and sampling activities, currently at 0.03 Sv. PMID- 15824590 TI - Health physics challenges involved with opening a "seventeen-inch" concrete waste vault. AB - This paper describes the various activities involved with opening a sealed legacy "Seventeen-inch" concrete vault and the health physics challenges and solutions employed. As part of a legacy waste stream that was removed from the former Hazardous Waste Management Facility at Brookhaven National Laboratory, the "Seventeen-inch" concrete vault labeled 1-95 was moved to the new Waste Management Facility for ultimate disposal. Because the vault contained 239Pu foils with a total activity in excess of the transuranic waste limits, the foils needed to be removed and repackaged for disposal. Conventional diamond wire saws could not be used because of facility constraints, so this project relied mainly on manual techniques. The planning and engineering controls put in place enabled personnel to open the vault and remove the waste while keeping dose as low as reasonably achievable. PMID- 15824591 TI - A history of the international commission on radiological protection. AB - Within twelve months of the discovery of x rays, papers appeared in the literature reporting adverse effects from high exposure. By the time of the First World War, several countries were proposing restrictions for the exposure of radiation workers. In 1925, the first International Congress of Radiology, held in London, considered the need for a protection committee, which it established at its second Congress in Stockholm in 1928. This paper traces the history of the development, by ICRP, of its policies and the personalities involved in their development from its inception up to the modern era. The paper follows the progress from the early controls on worker doses to avoid deterministic effects, through the identification of stochastic effects to the concerns about increasing public exposure. The key features of the Recommendations made by ICRP from 1928 up to the current 1990 version are identified. PMID- 15824592 TI - Radionuclides and trace metals in Canadian moose near uranium mines: comparison of radiation doses and food chain transfer with cattle and caribou. AB - Tissues from 45 moose and 4 cattle were collected to assess the health of country foods near uranium mines in northern Saskatchewan. Bone, liver, kidney, muscle and rumen contents were analyzed for uranium, radium-226 (226Ra), lead-210 (210Pb), and polonium-210 (210Po). Cesium-137 (137Cs), potassium-40 (40K), and 27 trace metals were also measured in some tissues. Within the most active mining area, Po in liver and muscle declined significantly with distance from tailings, possibly influenced by nearby natural uranium outcrops. Moose from this area had significantly higher 226Ra, 210Pb, 210Po, and 137Cs in some edible soft tissues vs. one control area. However, soil type and diet may influence concentrations as much as uranium mining activities, given that a) liver levels of uranium, 226Ra, and 210Po were similar to a second positive control area with mineral-rich shale hills and b) 210Po was higher in cattle kidneys than in all moose. Enhanced food chain transfer from rumen contents to liver was found for selenium in the main mining area and for copper, molybdenum and cadmium in moose vs. cattle. Although radiological doses to moose in the main mining area were 2.6 times higher than doses to control moose or cattle, low moose intakes yielded low human doses (0.0068 mSv y(-1)), a mere 0.3% of the dose from intake of caribou (2.4 mSv y( 1)), the dietary staple in the area. PMID- 15824593 TI - Testing prediction capabilities of an 131I terrestrial transport model by using measurements collected at the Hanford nuclear facility. AB - A model describing transport of 131I in the environment was developed by SENES Oak Ridge, Inc., for assessment of radiation doses and excess lifetime risk from 131I atmospheric releases from Oak Ridge Reservation in Oak Ridge, Tennessee, and from Idaho National Engineering and Environmental Laboratory in southeast Idaho. This paper describes the results of an exercise designed to test the reliability of this model and to identify the main sources of uncertainty in doses and risks estimated by this model. The testing of the model was based on materials published by the International Atomic Energy Agency BIOMASS program, specifically environmental data collected after the release into atmosphere of 63 curies of 131I during 2-5 September 1963, after an accident at the Hanford PUREX Chemical Separations Plant, in Hanford, Washington. Measurements of activity in air, vegetation, and milk were collected in nine counties around Hanford during the first couple of months after the accident. The activity of 131I in the thyroid glands of two children was measured 47 d after the accident. The model developed by SENES Oak Ridge, Inc., was used to estimate concentrations of 131I in environmental media, thyroid doses for the general population, and the activity of 131I in thyroid glands of the two children. Predicted concentrations of 131I in pasture grass and milk and thyroid doses were compared with similar estimates produced by other modelers. The SENES model was also used to estimate excess lifetime risk of thyroid cancer due to the September 1963 releases of 131I from Hanford. The SENES model was first calibrated and then applied to all locations of interest around Hanford without fitting the model parameters to a given location. Predictions showed that the SENES model reproduces satisfactorily the time-dependent and the time-integrated measured concentrations in vegetation and milk, and provides reliable estimates of 131I activity in thyroids of children. SENES model generated concentrations of 131I closer to observed concentrations, as compared to the predictions produced with other models. The inter-model comparison showed that variation of thyroid doses among all participating models (SENES model included) was a factor of 3 for the general population, but a factor of 10 for the two studied children. As opposed to other models, SENES model allows a complete analysis of uncertainties in every predicted quantity, including estimated thyroid doses and risk of thyroid cancer. The uncertainties in the risk-per-unit-dose and the dose-per-unit-intake coefficients are major contributors to the uncertainty in the estimated lifetime risk and thyroid dose, respectively. The largest contributors to the uncertainty in the estimated concentration in milk are the feed-to-milk transfer factor (F(m)), the dry deposition velocity (V(d)), and the mass interception factor (r/Y)dry for the elemental form of iodine (I2). Exposure to the 1963 PUREX/Hanford accident produced low doses and risks for people living at the studied locations. The upper 97.5th percentile of the excess lifetime risk of thyroid cancer for the most extreme situations is about 10(-4). Measurements in pasture grass and milk at all locations around Hanford indicate a very low transfer of 131I from pasture to cow's milk (e.g., a feed-to-milk transfer coefficient, F(m), for commercial cows of about 0.0022 d L(-1)). These values are towards the low end of F(m) values measured elsewhere and they are low compared to the F(m) values used in other dose reconstruction studies, including the Hanford Environmental Dose Reconstruction. PMID- 15824594 TI - Sensitivity analyses of the standards for the proposed Yucca Mountain repository- a review, evaluation, and commentary. AB - The standards and regulations for the proposed Yucca Mountain high level radioactive waste repository, which were developed and promulgated by the U.S. Environmental Protection Agency and the U.S. Nuclear Regulatory Commission, respectively, are complex and challenging. A major reason is that they are divided into three parts, an Individual Protection Standard, a Human Intrusion Standard, and multiple Ground Water Protection Standards. Because the individual parts are not fully integrated, the one that controls under a specific set of circumstances depends on the radionuclide being evaluated, its mechanisms of transport, its avenues of intake, and differences in the specified limits. Although the coefficients in Federal Guidance Report (FGR) No. 11 are being used to estimate the doses, other sources (for example, Title 10, CFR, Part 20, and/or FGR No. 13) may deserve consideration. Since the regulations specify that the reasonably maximally exposed individual is an adult, this leaves unanswered the estimated doses to other age groups, such as infants and adolescents. Summarized in this paper are comparisons of the dose coefficients for different age groups, as well as evaluations of the sensitivity of effective and organ dose estimates for adults, depending on the source of the coefficients. All the latter analyses were based only on the consumption of ground water. While the dose estimates are different, depending on the sources of the coefficients, this was not unexpected. What these evaluations demonstrate is the caution that must be exercised to ensure that a full range of considerations is taken into account in interpreting the outcome of the dose assessments being made with respect to the proposed repository. PMID- 15824595 TI - Measurements of neutron effective doses and attenuation lengths for shielding materials at the heavy-ion medical accelerator in Chiba. AB - The effective doses and attenuation lengths for concrete and iron were measured for the design of heavy ion facilities. Neutrons were produced through the reaction of copper, carbon, and lead bombarded by carbon ions at 230 and 400 MeV.A, neon ions at 400 and 600 MeV.A, and silicon ions at 600 and 800 MeV.A. The detectors used were a Linus and a Andersson-Braun-type rem counter and a detector based on the activation of a plastic scintillator. Representative effective dose rates (in units of 10(-8) microSv h(-1) pps(-1) at 1 m from the incident target surface, where pps means particles per second) and the attenuation lengths (in units of m) were 9.4 x 10(4), 0.46 for carbon ions at 230 MeV.A; 8.9 x 10(5), 0.48 for carbon ions at 400 MeV.A; 9.3 x 10(5), 0.48 for neon ions at 400 MeV.A; 3.8 x 10(6), 0.50 for neon ions at 600 MeV.A; 3.9 x 10(6), 0.50 for silicon ions at 600 MeV.A; and 1.1 x 10(7), 0.51 for silicon ions at 800 MeV.A. The attenuation provided by an iron plate approximately 20 cm thick (nearly equal to the attenuation length) corresponded to that of a 50-cm block of concrete in the present energy range. Miscellaneous results, such as the angular distributions of the neutron effective dose, narrow beam attenuation experiments, decay of gamma ray doses after the bombardment of targets, doses around an irradiation room, order effects in the multi-layer (concrete and iron) shielding, the doses from different targets, the doses measured with a scintillator activation detector, the gamma-ray doses out of walls and the ratio of the response between the Andersson-Braun-type and the Linus rem counters are also reported. PMID- 15824596 TI - Age-dependent dose assessment of 226Ra from bottled water intake. AB - Water may present a source of prolonged exposure to naturally occurring radionuclides. One of the most frequently occurring radionuclides in natural mineral and spring waters is 226Ra and its decay products. The contribution of drinking water to the total exposure is very small, at about 5% of the average effective dose attributable annually to natural background radiation, but that exposure contributes to the risk of adverse health consequences. In this study the mean values of 226Ra concentration determined in natural mineral and spring bottled waters range from 6 to 412 mBq L(-1), which is in accord with Croatian legislation. 226Ra effective doses per year from spring water consumption range up to 86 microSv, while 226Ra effective doses per year from mineral water consumption show much higher values. The highest 226Ra effective doses per year from mineral waters consumption, which are up to seven times higher than the dose recommended by WHO (100 microSv), were found in infants and teens. Based on this study, drinking of certain brands of bottled mineral water is not recommended for these age groups because assessed 226Ra effective doses per year exceed the recommended limits. From other research it is known that testosterone appears in elevated concentration during these life periods and affects bone calcification. Therefore, testosterone could affect the retention of 226Ra into the bone. To make more precise conclusions further research is necessary. Adults and especially elderly people are much less susceptible to the presence of 226Ra. According to the results obtained in this study, 226Ra effective doses per year assessed for these age groups were considerably lower (i.e., 10 microSv). PMID- 15824597 TI - Radiation exposure rate from 131I-treated hyperthyroid patients--a dynamic study, with data for up to 42 d post therapy. AB - Hyperthyroid patients treated with radioactive 131I are a potential source of external and internal exposure to family members and others in close contact with these patients. Information on the exposure rate from the patient on any day post administration of the dosage may be helpful when implementing an effective radiation safety or ALARA strategy for the family or members of the general public. Exposure rate measurements were completed on 78 out of 128 hyperthyroid patients participating in the study. Measurements were taken at 1 m, 0.6 m, and 0.3 m from the patient for eight different dose regimens and for up to 42 d post dose administration. The measured exposure rate was plotted against days post dose administration to demonstrate how quickly the exposure rate reduced with time. As anticipated, significant positive correlation was found between exposure rate at 1 m, 0.6 m, and 0.3 m for all radioactive dose regimens. No significant correlation was found between the external exposure rate reduction post-therapy and the 131I uptake at 2 or 24 h, prior to therapy. This work is a dynamic study that provides comprehensive external radiation exposure rate measurements in hyperthyroid patients post therapy dose administration and may serve as a database for radiation safety related decision-making. PMID- 15824598 TI - Does the severity of radionuclide-induced skeletal malignancy depend upon radiation dose? AB - The severity of effect as a function of skeletal radiation dose for induced skeletal malignancy was investigated among a relatively large group of beagle dogs singly injected as young adults with soluble radionuclide. Bone-seeking radionuclides considered were 239Pu, 226Ra, 228Ra, 224Ra, 228Th, and 90Sr. Variables included skeletal radiation dose, tumor growth rate, maximum tumor volume, sex of the animals, growth period of the tumor, degree of calcification of the tumor, skeletal location of the tumor, year of death, and occurrence of metastases. Except for a significant relationship between tumor volume and metastatic process and for growth rate and tumor volume, no significant dependence of any two of these factors could be established. It is concluded from available data on skeletal malignancies among beagles exposed to bone-seeking radionuclides that the severity of the disease is not dependent upon skeletal radiation dose. PMID- 15824599 TI - Comments about Potter's review of internal dosimetry. PMID- 15824604 TI - Pay-for-performance is coming to home care. PMID- 15824606 TI - Role of medications in the etiology and treatment of restless leg syndrome. PMID- 15824607 TI - Creating a POC from the initial assessment: tips for accurately completing other diagnoses and orders for discipline and treatments. PMID- 15824608 TI - M0830: emergent care. PMID- 15824610 TI - Lessons learned: from the other side of the nurse-patient/family relationship. AB - I have been a visiting nurse for more than 30 years. During that time, I have been a case manager, a supervisor, a team manager, and a wound ostomy continence specialist (WOCN). I have had all kinds of patients, including many seriously ill cancer patients. My colleagues consider me a knowledgeable, compassionate, and committed nurse. You would think I would be well prepared when serious illness came knocking on my door. I wasn't. PMID- 15824611 TI - Solving the mystery: the role of competency assessment in OASIS documentation. AB - Having an effective ongoing competency assessment program allows regulatory compliance for competency assessment, OASIS data collection, and information management, while enhancing existing quality management efforts. This article provides an overview of the competency assessment process, its regulatory underpinnings, and describes how this process fits into the existing performance and quality measurements within a home care organization. PMID- 15824612 TI - Oral manifestations of diabetes. AB - Periodontal disease, a microvascular complication of diabetes mellitus, is a group of infections and lesions affecting the tissues that form the attachment apparatus of a tooth or teeth. An individual with uncontrolled diabetes will have an increased risk of infection and abnormal healing time that will compromise the health of the oral cavity. A collaborative team approach between the home care clinician and dental professionals results in accurate patient assessment and education that results in better overall health outcomes. PMID- 15824614 TI - Medication safety: look-alike/sound-alike drugs in home care. AB - A new Joint Commission on Accreditation of Healthcare Organizations National Patient Safety Goal (NPSG) for 2005 is related to look-alike/sound-alike medications. This article reviews this new goal and what actions home care and hospice organizations need to undertake to meet the intent of this new requirement and provides insightful home care and hospice adaptations to the NPSG's general goals. Even if an agency is not Joint Commission-accredited, this information should be integrated into policies, procedures, and clinician education to avoid dangerous and costly medication errors. PMID- 15824616 TI - The clinician's role in Medicare Prospective Payment. Part 3-visits per episode: are you controlling utilization? PMID- 15824617 TI - Avoid sleepiness while driving. AB - Returning to the office after a long work day, Kristin, a hospice nurse, learns that the nighttime on-call nurse is ill. Kristin agrees to take call and hopes it will be a slow night. Now it is 2 am and Kristin has to travel to a client's home. Yawning repeatedly and driving over the rumble strip on the highway, she turns on the radio and opens the car window to the chill air, trying to shock herself into alertness. PMID- 15824618 TI - HPNA--leading the way. PMID- 15824621 TI - Neither snow, nor sleet, nor dark of night can stop a home care nurse. AB - With a blizzard leaving from 22 to 30 inches of snow in the Metro Boston area in January, and 8 more inches dumped two days later, home-visiting staff at the Visiting Nurse Association (VNA) of Boston switched into emergency mode. As one of the oldest and largest VNAs in the country, serving 15,000 patients annually in Greater Boston, the agency was ready. PMID- 15824624 TI - Did TCDD exposure or service in Southeast Asia increase the risk of cancer in air force Vietnam veterans who did not spray agent orange? AB - OBJECTIVE: We sought to examine cancer incidence in 1482 Air Force veterans who served in Southeast Asia (SEA) and who were not occupationally exposed to herbicides. METHODS: Cancer incidence between 1982 and 2003 was determined by record review and Cox proportional hazards models were used to estimate risk ratios across serum 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and years served in SEA categories. RESULTS: All sites cancer risk increased with TCDD (relative risk = 1.6, 95% confidence interval = 1.2-2.2). The risk of prostate cancer increased with years of SEA service but not with TCDD. TCDD and years of SEA service interacted with all sites cancer; the risk was greatest in those with the highest TCDD levels and the longest time served in SEA. CONCLUSIONS: These results suggest nonoccupational exposures to TCDD or other factors while in SEA may contribute to cancer risk in these veterans. PMID- 15824625 TI - The association of health status, worksite fitness center participation, and two measures of productivity. AB - OBJECTIVES: We sought to examine the associations between participation in a worksite fitness center and worker productivity. METHODS: A modified Work Limitations Questionnaire (WLQ) and employees' short-term disability claims were used as productivity measures with multivariate logistic regression models to control for health risk, age, gender, and work location. RESULTS: Nonparticipants in a worksite fitness center were more likely to report health-related work productivity limitations for time management (odds ratio [OR] = 1.62, 95% confidence interval [CI] = 1.09-2.41), physical work (OR = 1.58, CI = 1.03-2.43), output limitations (OR = 2.24, CI = 1.01-2.12), and overall work impairment (OR = 1.41, CI = 1.00-1.96) than fitness center participants. Fitness center participation also was associated with 1.3 days fewer short-term disability days per year per employee (P = 0.02) and 0.39 fewer health risks (P = 0.01). CONCLUSIONS: These results support the association of worksite fitness center participants with improved worker productivity and fewer short-term disability workdays lost. PMID- 15824626 TI - Workstyle and upper-extremity symptoms: a biobehavioral perspective. AB - OBJECTIVES: Workstyle has been proposed to help explain the link between ergonomic and psychosocial factors in work-related upper-extremity symptoms/disorders. This study investigated ergonomic factors, work demands, job stress, and workstyle on pain and functional limitations in computer users. METHODS: One hundred sixty-nine participants completed self-report ratings of job stress, ergonomic exposures, and workstyle at baseline. Three months, later ratings of pain and functional limitations were obtained. RESULTS: Multivariable logistic regression indicated that ergonomic exposure (odds ratio [OR] = 2.7, 95% confidence interval [CI] = 1.3-5.5), time spent at a computer (OR = 1.9, 95% CI = 0.95-3.6), and higher scores on a workstyle measure (2.4, 95% CI = 1.1-5.3) were independently associated with case status. Path analyses revealed that a model that included ergonomic exposure, work demands, and workstyle predicted pain and functional limitations at 3 months. CONCLUSIONS: Workstyle contributes to case definition and is predictive of future pain and functional limitations in office workers with upper extremity symptoms. PMID- 15824627 TI - Impact of the home indoor environment on adult asthma and rhinitis. AB - OBJECTIVE: We sought to study the combined effects of multiple home indoor environmental exposures in adult asthma and rhinitis. METHODS: We studied 226 adults with asthma and rhinitis by structured interviews and home assessments. Environmental factors included dust allergen, endotoxin and glucan concentrations, and indoor air quality (IAQ) variables. Outcomes included forced expiratory volume in 1 second (FEV1) percent predicted, Severity of Asthma Score (SAS), Short-Form (SF)-12 Physical Component Scale (PCS), and asthma Quality of Life (QOL) score. RESULTS: House dust-associated exposures together with limited IAQ variables were related to FEV1 % predicted (R = 0.24; P = 0.0001) and SAS (R = 0.18; P = 0.007). IAQ and limited dust variables were associated with SF-12 PCS (R = 0.15; P = 0.02), but not QOL (R = 0.13; P = 0.16). CONCLUSIONS: The home environment is strongly linked to lung function, health status, and disease severity in adult asthma and rhinitis. PMID- 15824628 TI - Availability of new drugs and Americans' ability to work. AB - OBJECTIVE: The objective of this work was the investigation of the extent to which the introduction of new drugs has increased society's ability to produce goods and services by increasing the number of hours worked per member of the working-age population. METHODS: Econometric models of ability-to-work measures from data on approximately 200,000 individuals with 47 major chronic conditions observed throughout a 15-year period (1982-1996) were estimated. RESULTS: Under very conservative assumptions, the estimates indicate that the value of the increase in ability to work attributable to new drugs is 2.5 times as great as expenditure on new drugs. CONCLUSIONS: The potential of drugs to increase employee productivity should be considered in the design of drug-reimbursement policies. Conversely, policies that broadly reduce the development and utilization of new drugs may ultimately reduce our ability to produce other goods and services. PMID- 15824629 TI - Low-level hexavalent chromium exposure and rate of mortality among US chromate production employees. AB - OBJECTIVE: Although numerous studies have reported an elevated lung cancer risk among chromium chemical production employees, few studies have focused on employees hired after major process changes and enhanced industrial hygiene controls were implemented. METHODS: This study examines the mortality experience of two post-change cohorts of chromate production employees constituting the current US chromium chemical industry. RESULTS: Mortality among chromium chemical workers generally was lower than expected on the basis of national and state specific referent populations. Lung cancer mortality was 16% lower than expected, with only three lung cancer deaths (3.59 expected). CONCLUSION: The absence of an elevated lung cancer risk may be a favorable reflection of the post-change environment. However, longer follow-up allowing an appropriate latency for the entire cohort will be needed to confirm this conclusion. PMID- 15824630 TI - Rethinking first response: effects of the clean up and recovery effort on workers at the world trade center disaster site. AB - OBJECTIVE: We sought to describe the physical and mental health effects of the cleanup and recovery effort on workers at the World Trade Center disaster site. METHODS: A mailed survey was sent to truck drivers, heavy equipment operators, laborers, and carpenters. It assessed work-related exposures and somatic and mental health symptoms. In one open-ended question, respondents shared any aspect of their experiences they wished; these 332 narrative responses were analyzed using qualitative techniques. RESULTS: Respondents reported suffering debilitating consequences of their work, including depression, drug use, and posttraumatic stress disorder. They felt poorly prepared to work in a disaster, lacked protective equipment and training, and felt overwhelmed by the devastation they faced. CONCLUSIONS: These workers' experiences were qualitatively similar to the experiences of the first responders. To protect workers in the future, the focus on preparing "first" responders should be reconsidered more broadly. PMID- 15824631 TI - Occupation and risk of pancreatic cancer: a population-based case-control study in iowa. AB - OBJECTIVE: Previous epidemiological studies have inconsistently linked various occupations and industries to pancreatic cancer risk. METHODS: We analyzed data from a population-based case-control study conducted in Iowa involving 376 histologically confirmed incident pancreatic cancer cases and 2434 control subjects. RESULTS: A significantly increased risk was observed among men who worked in the following industries: chemical and allied products, transportation, and elementary and secondary schools. Increased risks also were observed in men who were employed as truck drivers; railroad brake, signal, and switch operators; purchasing agents and buyers; teachers; insurance agents; and retail supervisors. Among women, a significantly increased risk of pancreatic cancer was found for employment in furniture and home furnishing stores, and a borderline significantly increased risk among textile sewing machine operators and tenders. CONCLUSIONS: Working in several occupations and industries was associated with an increased risk of pancreatic cancer in this study, and these associations warrant further investigation. PMID- 15824632 TI - Integrating qualitative research into occupational health: a case study among hospital workers. AB - OBJECTIVE: We sought to better use qualitative approaches in occupational health research and integrate them with quantitative methods. METHODS: We systematically reviewed, selected, and adapted qualitative research methods as part of a multisite study of the predictors and outcomes of work-related musculoskeletal disorders among hospital workers in two large urban tertiary hospitals. RESULTS: The methods selected included participant observation; informal, open-ended, and semistructured interviews with individuals or small groups; and archival study. The nature of the work and social life of the hospitals and the foci of the study all favored using more participant observation methods in the case study than initially anticipated. CONCLUSIONS: Exploiting the full methodological spectrum of qualitative methods in occupational health is increasingly relevant. Although labor-intensive, these approaches may increase the yield of established quantitative approaches otherwise used in isolation. PMID- 15824633 TI - The relation of occupational organic solvent exposure to symptom reporting in a sample of white and Chinese midlife women. AB - OBJECTIVE: This study examined the relation of occupational solvent exposure to menopausal and other symptoms in midlife women. METHODS: We conducted a cross sectional study of 480 Chinese and 494 white women, aged 40-55 years, in Northern California. Levels of exposure to organic solvents (none, low, medium, or high) were assigned to each current job using a job-exposure matrix. RESULTS: A lower proportion of women with low occupational organic solvent exposure reported hot flashes or night sweats than working women with no solvent exposure (adjusted prevalence odds ratio [APOR] = 0.48, 95% confidence interval [CI] = 0.19-1.21). A greater proportion of women with high solvent exposure reported forgetfulness than women with no exposure (APOR = 2.51, 95% CI = 1.12-5.63). CONCLUSIONS: Some symptom reporting in midlife women was related to their occupational organic solvent exposure. PMID- 15824634 TI - Depression and occupational injury: results of a pilot investigation. AB - OBJECTIVE: Even mild clinical depression can cause decreased vigilance, attention span, increased irritability, and insomnia-all well-known precursors to occupational injury. This pilot project explores the relationship between occupational injury and depression. METHOD: One hundred twenty-one individuals with recent work-related injuries and 140 without work-related injuries completed a self-administered depression screening instrument (PHQ-9). We compared the two groups using bivariate analyses. The impact of depression on injury was examined using logistic regression analysis controlling for employment history, marital status, age, and sex. RESULTS: Overall, injured workers in this study were not more likely to be depressed than a comparison group of uninjured workers. However, injured women had significantly higher depression scores than non injured women (P = 0.04); no such difference was found for men. CONCLUSIONS: These data suggest that depression may serve as a precursor to occupational injury for women. PMID- 15824635 TI - Work-related recovery expectations and the prognosis of chronic low back pain within a workers' compensation setting. AB - OBJECTIVE: We examined the association between work-related recovery expectations and return-to-work in patients with chronic back pain. METHODS: A prospective cohort of workers receiving time-loss benefits for back pain of at least 6 weeks' duration was studied. Workers completed a battery of measures, including a work related recovery expectations questionnaire. Outcomes included surrogate indicators of timely return-to-work (days until suspension of time-loss benefits) and recovery (claim closure) censored at 1 year. Analysis included multivariable Cox and logistic regression. RESULTS: The sample was predominantly male (63%), with a mean age of 42 years. Positive work-related recovery expectations predicted a 26% faster suspension of time-loss benefits (95% confidence interval = 8-40%) and explained 7% of the variation in this outcome. CONCLUSION: Recovery expectations provide important information for predicting time to return-to-work, as measured through suspension of time-loss benefits (R approximately 7%) in patients with chronic back pain. PMID- 15824636 TI - Hot water for handwashing--where is the proof? PMID- 15824639 TI - The academic trauma center is a model for the future trauma and acute care surgeon. AB - BACKGROUND: Strategies to prevent the extinction of the trauma surgeon have focused on increasing the operative potential by including nontrauma general surgery emergencies. Although providing comprehensive emergent surgical care by the trauma service may seem novel, our institution has embraced this concept for the past 25 years. Recent discussions on the future of trauma surgery stimulated us to review our experience as a possible model for the future trauma and acute care surgeon. METHODS: We reviewed operative logs for 2002 and 2003 at our urban academic Level I trauma center. Six surgeons participate equally in call that covers trauma and nontrauma general surgical, thoracic, and vascular emergencies. Cases were classified as trauma, emergent, urgent, or according to the patient's clinical condition. The primary procedure for each operation was classified according to the American Board of Surgery Case Reporting System. RESULTS: We performed 4,082 operations during the study period, of which 8% were trauma, 11% were emergent, 40% were urgent, and 41% were elective. Abdominal and alimentary procedures accounted for 53% of all operations. Vascular, thoracic, and head and neck procedures accounted for 22%, 14%, and 9% of procedures, respectively. CONCLUSION: To resurrect our discipline, we must reclaim and expand our operative potential and be relieved of our excessive night and weekend burden of serving as housestaff for the neurosurgeons, orthopedic surgeons, and interventional radiologists. The trauma surgeon can effectively manage trauma and acute care surgery emergencies including thoracic and vascular conditions. Education of the future trauma and acute care surgeon must include specialty training in thoracic and vascular surgery. PMID- 15824640 TI - The future of trauma surgery--a perspective. PMID- 15824641 TI - Dollars and sense: attributing value to a level I trauma center in economic terms. AB - BACKGROUND: Reliable, accurate, program-specific data for hospital product lines are often difficult to obtain. The purpose of this study was to determine the impact that trauma center status has on hospital net income when compared with other traditional hospital product lines and services. METHODS: Over a 3-year period, financial data were collected for 16 payor classes: 8 major payors for all injury diagnoses, in-patient and out-patient. These data were analyzed by total charges, total direct costs, contribution margin, and net income. A key assumption of this strategy was that although injury patients are treated at most hospitals, only trauma center status allows access to patients and provision of services that can contribute significantly to the bottom line. RESULTS: Over the 3-year period, the contribution margin increased for trauma patients (excluding Level I trauma), Level I trauma patients, and the combined population of trauma patients. The most significant portion of the increase resulted from patients seen as a result of trauma center status. CONCLUSION: We conclude that, for our institution, the investment in resources necessary to achieve and maintain trauma center status makes economic sense in that the trauma program contributes favorably to hospital net revenue. PMID- 15824642 TI - The invisible trauma patient: emergency department discharges. AB - BACKGROUND: As the malpractice and financial environment has changed, injured patients evaluated by the trauma team and discharged from the emergency department (ED) are now commonplace. The evaluation, care, and disposition of this population has become a significant workload component but is not reported to accrediting organizations and is relatively invisible to hospital administrators. Our objective was to quantify and begin to qualify the evolving picture of the trauma ED discharge population as a work component of trauma service function in an urban, Level I trauma center with an aeromedical program. METHODS: Trauma registry (contacts, mechanism, transport, injuries, and disposition) and hospital databases (ED closure, occupancy rates) were queried for a 5-year period (1999-2003). Trend analysis provided statistical comparisons for questions of interest. RESULTS: During the 5-year study period, the total number of trauma contacts rose by 18.1% (2,220 in 1999 vs. 2,622 in 2003; trend p < 0.05). This increase in total contacts was not a manifestation of an increase in admissions (1,672 in 1999 vs. 1,544 in 2003) but rather a reflection of a marked increase in patients seen primarily by the trauma team and discharged from the ED (473 in 1999 vs. 1,000 in 2003; trend p < 0.05). These ED discharge patients were increasingly transported by helicopter (12.3% in 1999 vs. 29.2% in 2003; trend p < 0.05) and less frequently from urban areas (57.1% in 1999 vs. 48.1% in 2003; trend p < 0.05) over the course of the study period. Average injury severity of this group increased over the study period (Injury Severity Score of 2.7 +/- 0.1 in 1999 vs. 3.3 +/- 0.1 in 2003; trend p < 0.05). ED length of stay for this group increased 19.8% over the study period (trend p < 0.05), averaging nearly 5 hours in 2003. CONCLUSION: The total number, relative percentage, and injury severity of patients evaluated by the trauma team and discharged from the ED has significantly increased over the last 5 years, representing nearly 5,000 patient care hours in 2003. Systems to care for these patients in a cost- and resource-efficient fashion should be put in place. The impact of this growing population of patients on the workload of the trauma center should be recognized by accrediting agencies, hospital administration, and Emergency Medical Services. PMID- 15824643 TI - How does casualty load affect trauma care in urban bombing incidents? A quantitative analysis. AB - BACKGROUND: The aim of this modeling study was to examine how casualty load affects the level of trauma care in multiple casualty incidents and to define the surge capacity of the hospital trauma assets. METHODS: The disaster plan of a U.S. Level I trauma center was translated into a computer model and challenged with simulated casualties based on 223 patients from 22 bombing incidents treated at an Israeli hospital. The model assigns providers and facilities to casualties and computes the level of care for each critical casualty from six variables that reflect the composition of the trauma team and access to facilities. RESULTS: The model predicts a sigmoid-shaped relationship between casualty load and the level of care, with the upper flat portion of the curve corresponding to the surge capacity of the trauma assets of the hospital. This capacity is 4.6 critical patients per hour using immediately available assets. A fully deployed disaster plan shifts the curve to the right, increasing the surge capacity to 7.1. Overtriage rates of 50% and 75% shift the curve to the left, decreasing the surge capacity to 3.8 and 2.7, respectively. CONCLUSION: This model defines the quantitative relationship between an increasing casualty load and gradual degradation of the level of trauma care in multiple casualty incidents, and defines the surge capacity of the hospital trauma assets as a rate of casualty arrival rather than a number of beds. The study demonstrates the value of dynamic computer modeling as an important tool in disaster planning. PMID- 15824644 TI - Computed tomographic scanning and selective laparoscopy in the diagnosis of blunt bowel injury: a prospective study. AB - BACKGROUND: We prospectively evaluated whether computed tomographic (CT) scanning and selective laparoscopy (LP) for the diagnosis of blunt bowel injury (BBI) could prevent nontherapeutic laparotomy and delayed diagnosis. METHODS: Between April 1994 and May 2002, hemodynamically stable patients suspected of having BBI were enrolled in this study. Patients with hemodynamic instability or solid organ injuries with hemoperitoneum were excluded. All patients underwent a physical examination and contrast CT scanning at admission and once again approximately 12 hours (range, 6-24 hours) after admission. LP was performed under general anesthesia in patients who had local peritoneal signs and indirect CT signs (bowel thickening or isolated intraperitoneal fluid) or in whom abdominal pain or tenderness increased or intraperitoneal fluid increased on the repeat CT scan. The indications for a celiotomy were diffuse peritonitis, pneumoperitoneum on the abdominal CT scan, or bowel perforation visible on LP. RESULTS: During the study period, 399 of 1,074 patients admitted for blunt torso injuries were enrolled in this study. Eleven patients underwent emergency celiotomy and 11 underwent LP immediately after admission to the emergency department. One nontherapeutic laparotomy was performed among the patients who underwent celiotomy. The LPs revealed seven bowel perforations and one mesenteric laceration. After a repeat CT scan, three and seven of the patients underwent laparotomy and LP, respectively. Four bowel perforations were found by LP. The remaining 198 patients were treated conservatively, and no complications related to a delayed BBI diagnosis occurred. CONCLUSION: CT scanning and selective LP can prevent nontherapeutic laparotomy and delayed diagnosis in patients with suspected BBI. PMID- 15824645 TI - Iliac vein injuries in hemodynamically unstable patients with pelvic fracture caused by blunt trauma. AB - BACKGROUND: Major pelvic venous injuries secondary to blunt trauma can be a difficult problem in diagnosis and management. This study aimed to elucidate the clinical significance of iliac vein injuries demonstrated by venography in patients with blunt pelvic injuries who remained unstable even after transcatheter arterial embolization (TAE). METHODS: We reviewed the records of 72 patients with unstable pelvic fracture who presented with shock at our center after blunt trauma from 1999 through 2003. The average Injury Severity Score was 34.3 in this study population. RESULTS: TAE was the first method of choice to control bleeding from pelvic fracture in 61 patients. Thirty-six patients recovered from shock after TAE. Eighteen of 25 who did not recover from shock died. In 11 of these 25, transfemoral venography with a balloon catheter was performed, revealing significant venous extravasation in 9: common iliac vein in 5, internal iliac vein in 3, and external iliac vein in 1. The average Injury Severity Score of patients with iliac vein injury was 45.8. Treatments for venous injuries were laparotomy for hemostasis (n = 1, survivors = 0), retroperitoneal gauze packing (n = 3, survivors = 1), and endovascular stent placement (n = 3, survivors = 3). Two patients suffered from cardiac arrest before treatment for venous injury. External fixations were performed after TAE according to fracture type. CONCLUSION: The iliac vein injury is the principal cause of hemorrhagic shock in some patients with unstable pelvic fractures after blunt trauma. Venography is useful for identifying iliac vein injuries. PMID- 15824646 TI - Differential effects of anesthetics on endotoxin-induced liver injury. AB - BACKGROUND: The liver is both a source and a target of inflammatory and anti inflammatory mediators during sepsis. The oxidative stress proteins inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1) are upregulated in the liver during sepsis but have opposite roles. Upregulation of HO-1 has hepatoprotective effects, whereas iNOS has injurious effects to the liver. Although recent studies indicate that ketamine anesthesia has anti-inflammatory effects during sepsis, the effects of other anesthetics are unknown. We hypothesized that ketamine, but not isoflurane, would attenuate lipopolysaccharide (LPS)-induced liver injury through differential modulation of iNOS and HO-1. METHODS: Adult rats were given no anesthesia (saline), continuous isoflurane inhalation, or intraperitoneal ketamine (70 mg/kg). One hour later, saline or LPS (20 mg/kg intraperitoneally) was given for 5 hours. Rats were killed, serum prepared for determination of hepatocellular enzymes, and the liver assessed for iNOS and HO-1 by Western immunoblot. RESULTS: LPS significantly increased serum aspartate aminotransferase levels, iNOS, and HO-1 immunoreactivity in the liver. Ketamine but not isoflurane attenuated LPS-induced liver injury, upregulated HO-1, and downregulated iNOS. CONCLUSION: These data indicate that anesthetics differ in their effects on the liver in a rat model of sepsis with LPS. Ketamine has hepatoprotective effects against LPS-induced liver injury that appear to be mediated, at least in part, by differential modulation of the oxidative stress proteins iNOS and HO-1. Thus, ketamine may be the anesthetic agent of choice for septic patients requiring anesthesia. PMID- 15824647 TI - The use of neuromuscular blocking agents to facilitate prehospital intubation does not impair outcome after traumatic brain injury. AB - BACKGROUND: Several studies have demonstrated that the success rate of prehospital intubation is improved by the use of neuromuscular blocking agents (NMBAs). However, a recent study has reported that prehospital intubation with NMBAs worsens outcome after traumatic brain injury (TBI). We sought to determine the effect of the use of NMBAs to facilitate prehospital intubation on outcome after TBI. METHODS: All patients admitted to our Level I trauma center with a head Abbreviated Injury Scale score >/= 3 were identified by the trauma registry (January 1998-June 2003). Patient records were matched with prehospital databases. Patients were further stratified on the basis of prehospital Glasgow Coma Scale (GCS) score into mild (GCS score of 14/15), moderate (GCS score of 9 13), and severe (GCS score < 9) TBI. Outcome included mortality and good outcome (survival to discharge with a GCS score of 14/15). RESULTS: There were 3,052 patients who were identified as having been transported directly from the field. Complete prehospital data were available for 2,012 patients (66%). Of these, 920 were mild TBI (intubation rate, 17.4%), 293 moderate TBI (intubation rate, 57.7%), and 799 severe TBI (intubation rate, 95%). Overall, 72% of intubated patients received NMBAs. There were no significant differences in demographics or injury severity between the groups. Patients not receiving NMBAs were more likely to be hypotensive and have prehospital cardiopulmonary resuscitation (p = 0.001). The unadjusted mortality for the patients intubated with NMBAs was 25% versus 37% for those not receiving NMBAs (p < 0.001). When adjusted for confounding variables, patients intubated with NMBAs were more likely to survive (odds ratio, 0.63; 95% confidence interval, 0.41-0.97; p = 0.04) and have a good outcome (odds ratio, 1.7; 95% confidence interval, 1.2-2.6; p = 0.006) than those in the no NMBA group. CONCLUSION: The use of NMBAs to facilitate prehospital intubation improves outcome for patients with TBI. The value of prehospital intubation for TBI remains to be determined; however, any trial evaluating nonintubation for TBI must be compared with NMBA-facilitated intubation to be valid. PMID- 15824648 TI - Subsequent development of thrombocytopenia and coagulopathy in moderate and severe head injury: support for serial laboratory examination. AB - BACKGROUND: Patients with moderate and severe traumatic brain injury (TBI) are at risk for secondary brain insults such as thrombocytopenia and coagulopathy. This study assessed the development of thrombocytopenia and coagulopathy at admission and within the subsequent 72 hours after TBI. METHODS: Blunt trauma patients with moderate or severe TBI and an extracranial Abbreviated Injury Scale score less than 3 were reviewed. Data collection included initial and subsequent prothrombin time, partial thromboplastin time, and platelet values. RESULTS: On initial evaluation, thrombocytopenia was present in 14% and coagulopathy in 21% of patients. By the third day, thrombocytopenia and coagulopathy increased to 46% and 41%, respectively. Of patients who died, 67% had thrombocytopenia and 62% had coagulopathy. CONCLUSION: Patients with moderate and severe TBI are at risk for thrombocytopenia and coagulopathy, not only at admission but also on subsequent laboratory examination. Repeat laboratory evaluation is warranted even if initial results are normal in this population. PMID- 15824649 TI - Evaluation of the applicability, efficacy, and safety of a thromboembolic event prophylaxis guideline designed for quality improvement of the traumatically injured patient. AB - BACKGROUND: Thromboembolic events (TE) such as deep venous thrombosis (DVT) and pulmonary embolism (PE) are common after trauma. Our Trauma Practice Management Committee developed an evidence-based DVT/PE prophylaxis guideline using a modified Delphi approach to standardize care and reduce TE rates. Our objective was to evaluate the applicability, efficacy, and safety of this guideline in the traumatized patient, especially those admitted first to the intensive care unit (ICU). METHODS: We developed a risk-stratified DVT/PE prophylaxis guideline incorporating specific injuries, pertinent history, and physiologic parameters, favoring aggressive therapy in those at highest risk of dying from a PE. We prospectively collected data using this guideline in all patients admitted to the trauma or orthopedic-trauma services that were expected to stay for more than 48 hours (March-December 2003). Comparison was made with historical controls. Data collected included DVT, PE, prophylaxis level chosen, inferior vena cava filters, admission service and location, TRISS scores, length of stay, outcomes, adverse events, and specific risk factors. RESULTS: TE rates after implementation of the guideline were lower than historical controls for all patients (1.9% vs. 1.0%, p = 0.059) and for patients admitted first to the ICU (6.3% vs. 2%, p = 0.018). Completed sheets were collected for 46% of the targeted population. No bleeding events caused by guideline anticoagulation were noted, and one death occurred after inferior vena cava filter placement. Nine of the 12 TEs in the treatment group were in patients with spine or closed-head injury, delaying chemical prophylaxis. CONCLUSION: Form-based, risk-adjusted prophylaxis against TE leads to lower TE rates in a general and orthopedic ICU trauma population. Protocol compliance should be enforced. PMID- 15824650 TI - Kupffer cell ablation improves hepatic microcirculation after trauma and sepsis. AB - BACKGROUND: Macrophages undergo maladaptive alterations after trauma. In this study, we assessed the role of Kupffer cells in hepatic microcirculatory response to endothelin-1 (ET-1) after femur fracture (FFx) and cecal ligation and puncture (CLP). METHODS: Sprague-Dawley rats (200-300 g) underwent sham, FFx, CLP, or FFx + CLP. To ablate Kupffer cells, group 1 animals were treated with gadolinium chloride, and group 2 animals received saline. Hepatic microcirculation was assessed by intravital microscopy. Liver mitochondrial redox state and tissue oxygen (tPo2) were determined by NADH and ruthenium fluorescence, respectively. Liver damage was estimated by alanine aminotransferase levels. Differences were assessed using analysis of variance followed by Student-Newman-Keuls post hoc test. RESULTS: After 10 minutes of ET-1, CLP and FFx + CLP caused significant reduction in hepatic perfusion index (2.5-fold and 5-fold vs. sham, p < 0.05, respectively), redox state (36% and 45% vs. sham, p < 0.01, respectively), tPo2 (10% and 12% vs. sham, p < 0.05, respectively), and more liver damage compared with sham and FFx-treated animals. Kupffer cell depletion restored microcirculation, redox state, and tPo2 and abrogated hepatocellular damage. CONCLUSION: Kupffer cells contribute directly to hepatic microcirculatory dysfunction and liver injury after inflammatory stress. Furthermore, Kupffer cell depletion ameliorates the microcirculatory perturbations of trauma and sepsis. Thus, modulation of Kupffer cell response may prove beneficial. PMID- 15824651 TI - Clinically relevant osmolar stress inhibits priming-induced PMN NADPH oxidase subunit translocation. AB - BACKGROUND: The plasma membrane NADPH oxidase is responsible for the external generation of superoxide by neutrophils (polymorphonucleocytes [PMNs]). The oxidase is a multicomponent enzyme, active only when all subunits are translocated to and assembled at the membrane. We have recently demonstrated that platelet-activating factor (PAF) priming of PMNs translocates the cytosolic p67 subunit to the membrane position. Osmolar stress attenuates PAF priming of the oxidase. Consequently, we hypothesized that clinically relevant osmolar stress inhibits PAF priming-induced p67 translocation. METHODS: Isolated human PMNs were incubated at 37 degrees C for 5 minutes in buffer or 180 mmol/L hypertonic saline (HTS) followed by 3 minutes of incubation with or without 2 mumol/L PAF (resting, PAF, HTS, and HTS-PAF). Digital microscopy was used to determine p67 location in whole PMNs. Subcellular fractions were prepared and membrane translocation of p67 determined by protein electrophoresis. Resting cytosol fractions were immunodepleted of p67 and NADPH oxidase activity measured using p67-deficient sodium dodecyl sulfate cell-free oxidase assays: resting, PAF, or HTS-PAF membrane (1 mug) was combined with immunodepleted resting cytosol (25 mug). RESULTS: By all methodologies, PAF stimulated translocation of p67 to the PMN membrane and this translocation was prevented by osmolar stress (HTS-PAF). In cell-free oxidase assays, the membrane content of p67 after PAF stimulation was increased sufficiently to induce oxidase activity, whereas resting and HTS-PAF membrane did not (0.1 +/- 0.02, 0.23 +/- 0.04, and 0.14 +/- 0.04, respectively, p < 0.01) (resting versus HTS-PAF, no difference). CONCLUSION: PAF priming of the PMN oxidase involves translocation of p67 to the plasma membrane. Clinically relevant osmolar stress with hypertonic saline prevents this PAF-induced translocation of the p67 oxidase subunit. This finding provides new insight into the mechanisms responsible for osmolar control of PMN functional responses. PMID- 15824652 TI - Life after 80 hours: the impact of resident work hours mandates on trauma and emergency experience and work effort for senior residents and faculty. AB - BACKGROUND: The purpose of this study was to evaluate the impact of work hours mandates on (1) senior resident patient exposure and operating experience in trauma and emergency surgery and (2) faculty work effort. METHODS: We measured resident and faculty work on the trauma and emergency surgery services at our Level I trauma center during two comparable 6-month periods. Period 1 (July 1 December 31, 2002) had no call restrictions, separate trauma and emergency service resident call, and some overlap of faculty call responsibilities. Period 2 (July 1-December 31, 2003) had resident work hours compliance and complete integration of resident and faculty trauma and emergency call. Work hours were measured by surveys for faculty and residents. All data were collected prospectively. RESULTS: Resident exposure to trauma patients was similar during both time periods. Emergency surgery admissions declined during period 2; however, intensive care unit admissions increased. The number of operations performed by senior residents did not change; however, there was a shift in the median number of emergency surgery cases to more senior residents. Faculty work hours increased slightly despite a decrease in faculty call. CONCLUSION: Work hours compliance resulted in a 50% reduction in senior resident call and a 19% decrease in their work hours with no significant change in trauma/emergency patient care exposure or operative case load. Service call amalgamation reduced faculty call by 21% but did not result in a corresponding change in work hours or productivity. PMID- 15824653 TI - Long-term posttraumatic stress disorder persists after major trauma in adolescents: new data on risk factors and functional outcome. AB - BACKGROUND: Little is known about long-term psychological outcomes after major trauma in adolescents, a leading cause of death and preventable morbidity in this age group. A prospective epidemiologic study was conducted to examine quality of life (QoL) and posttraumatic stress disorder (PTSD) outcomes in injured adolescents. The specific objectives of the present report are to describe long term PTSD and to identify risk factors for long-term PTSD and the impact of PTSD on QoL. METHODS: Between April 26, 1999, and November 13, 2002, 401 eligible trauma patients aged 12 to 19 years triaged to five participating trauma center hospitals in a regionalized trauma system were enrolled in the study. The admission criteria for patients were as follows: age 12 to 19 years and injury diagnoses excluding severe traumatic brain injury or spinal cord injury. PTSD was diagnosed with the Impact of Events Scale-Revised; scores of 24+ were used to diagnose PTSD. QoL after trauma was measured using the Quality of Well-Being scale, a sensitive and well-validated functional index (range: 0 = death to 1.000 = optimum functioning). Patient outcomes were assessed at discharge and at 3, 6, 12, 18, and 24 months after discharge. RESULTS: The rate of long-term PTSD was 27%, with high rates over the follow-up period. Risk factors for long-term PTSD were perceived threat to life (odds ratio [OR], 2.2; p < 0.001.); death of a family member at the scene (OR, 4.7; p < 0.001); no control over injury event (OR, 1.7; p < 0.05), and violence-related injury (OR, 2.2; p < 0.05). PTSD in adolescents was significantly and strongly associated with female gender, older age, low socioeconomic status, drug and alcohol abuse, and other adolescent behavioral problems. Long-term PTSD was also associated with marked and significant QoL deficits throughout the 24-month follow-up interval. CONCLUSION: High rates of long-term PTSD persist after major trauma in adolescents. Injury events such as perceived threat to life and control over the event are strongly associated with PTSD risk. Prolonged PTSD severely impacts QoL outcomes. Early identification and treatment of risk factors for long-term PTSD will be important to improve outcomes in injured adolescents. PMID- 15824654 TI - Epidemiology of suicide and the impact on Western trauma centers. AB - BACKGROUND: Suicide represents the eleventh leading cause of death in the United States. Most suicides are a result of self-directed injury and are commonly treated in trauma centers. The purpose of this article is to characterize the epidemiology of suicide in the Western Mountain states and to describe the impact of these injuries on trauma centers. METHODS: Data were accumulated from the National Trauma Data Bank, the American Association of Suicidology, and the Centers for Disease Control and Prevention Web-based Injury Statistics Query and Reporting System. Variables analyzed included the epidemiology of suicide in relation to regional location, age, gender, mechanism of injury, mortality, disposition, Total Functional Independence Measure (TFIM) score, and length of stay in the intensive care unit versus the hospital floor. Statistical analysis was performed on select data points to demonstrate relative significance. Several comparisons were made among intentional injuries directed at oneself, at others, and unintentional injuries with respect to the above variables. RESULTS: The Western region of the country has maintained the highest suicide rates since 1990, with 8 of the 10 Western Mountain states having rates twice as high as the national average. Approximately 88% of suicides presenting to trauma centers involve firearms, stabbings, or falls, usually causing injuries that lead to significant morbidity and mortality. Suicide attempts represent 2% of all admissions to trauma centers, with two thirds of these patient dying while in the hospital. Average length of stay in the intensive care unit and on hospital floors was 2.2 and 5.2 days, respectively, which was higher than the average length of stay of either unintentional injuries or intentional injuries directed at others. CONCLUSION: The Western Mountain states represent an endemic region for suicide. Because of the violent nature of these injuries, they often require significant medical care and finances usually provided by state governments. Suicides also contribute to the morbidity and mortality in a disproportionate manner in comparison with motor vehicle crashes and homicides. In conclusion, the Western region bears a larger burden than other regions in caring for this subset of patients with respect to medical and financial resources. Increased public awareness and preventative measures can aid in reducing not only the overall incidence of suicide but also the patient load and costs that are incurred on this country's health care system. PMID- 15824655 TI - Institutional practice guidelines on management of pelvic fracture-related hemodynamic instability: do they make a difference? AB - BACKGROUND: The management of patients with hemodynamic instability related to pelvic fracture is a major challenge, with high morbidity and mortality. Evidence based institutional practice guidelines (PG) were developed as a strategy to optimize the care of these patients. The aims of this study were to evaluate the adherence to the new PG and compare the outcomes before and after their implementation. METHODS: Major blunt trauma patients (Injury Severity Score [ISS] > 15) with hemodynamic instability (initial base deficit > 6 mEq/L or received > 6 units of packed red blood cells [PRBCs] during the first 12 hours) related to pelvic fracture were investigated. Patients presenting with ongoing bleeding from other regions or with severe head injury (Glasgow Coma Scale score < 9) were excluded. The pre-PG group (n = 17) were patients managed during the 18 months ending on December 31, 2001. The post-PG group (n = 14) consisted of patients managed during the subsequent 18 months. Demographics, ISS, shock severity, resuscitation, and outcome data were prospectively collected. The adherence to the key steps of PG was evaluated retrospectively in the pre-PG and prospectively in the post-PG group, including abdominal clearance (AC) with diagnostic peritoneal aspiration/lavage or ultrasound (<15 minutes), noninvasive pelvic binding (PB) (<15 minutes), pelvic angiography (PA) (<90 minutes after admission), and minimally invasive orthopedic fixation (MIOF) (<24 hours). Data are presented as mean +/- SEM or percentages. RESULTS: The pre-PG and post-PG groups were similar regarding age (40 +/- 4 years vs. 42 +/- 6 years), gender (both 71% male), ISS (39 +/- 3 vs. 37 +/- 4), admission base deficit (9 +/- 1 vs. 10 +/- 1) admission systolic blood pressure (116 +/- 7 vs. 112 +/- 6 mm Hg), Glasgow Coma Scale score (12 +/- 1 vs. 12 +/- 1), and PRBC transfusion in the first 12 hours (9 +/- 2 U vs. 9 +/- 2 U). The adherence to the guidelines in the post-PG period was as follows: AC, 100%; PB, 86% (p < 0.05 based on t test or chi test); PA, 93% (p < 0.05 based on t test or chi test); and MIOF, 86%. In the pre PG period, adherence to the guidelines was as follows: AC, 65%; PB, 0%; PA, 30%; and MIOF 52%. In the post-PG period, the 24-hour PRBC transfusion decreased from 16 +/- 2 U to 11 +/- 1 U and the mortality decreased from 35% to 7% (p < 0.05 based on t test or chi test for both). CONCLUSION: The adherence to the PG as a reflection of optimal management was significantly improved. PG focusing particular on timely hemorrhage control reduced the 24-hour transfusion requirements and the mortality rate in the post-PG group. PMID- 15824656 TI - Unsafe at any age: a retrospective review of all-terrain vehicle injuries in two level I trauma centers from 1995 to 2003. AB - BACKGROUND: All-terrain vehicles (ATVs) are popular recreational and utility vehicles. In 1984, Cogbill published an article regarding three-wheelers. These are no longer manufactured, but the injury and death rate with four-wheeled ATVs is high and disproportionately affects young riders. METHODS: We conducted a retrospective review at two Level I trauma centers from January 1994 to April 2003. Statistical analysis was performed using the SAS V8.2 program. Values of p < 0.05 were significant. RESULTS: Two hundred eight patients were identified. There were no differences identified in demographics, mechanism, types of injury, Injury Severity Score (ISS), or Glasgow Coma Scale (GCS) score. Seventy-five percent were male and 84% were white. The mean age was 23 +/- 13 years. The average ISS was 12.3 +/- 9 and the mean GCS score was 13.1 +/- 3.7. Injury mechanisms were loss of stability (33%), separation of rider from ATV (32%), and ATV versus stationary object (27%). ISS for ages 12 to 15 years was significantly higher than for other ages (14.5 vs. 11.5, p = 0.04, Wilcoxon rank sum test) and included more major head injuries (40.4% vs. 21.8%, p = 0.09, Wilcoxon rank sum test). They experienced fewer spinal fractures (3.9% vs. 15.4%, p = 0.03) and pelvic injuries (0% vs. 9%, p = 0.02, Wilcoxon rank sum test). The GCS score in this group was lower (12.3 vs. 13.4, p = 0.03, Wilcoxon rank sum test). CONCLUSION: Adolescent ATV riders have more severe injuries and more head injuries than other age groups. Prevention efforts should target this group. PMID- 15824657 TI - Laparoscopy is sufficient to exclude occult diaphragm injury after penetrating abdominal trauma. AB - BACKGROUND: Occult diaphragm injury after penetrating thoracoabdominal injury can be difficult to diagnose and can remain occult for months to years. Delayed diagnosis is associated with the risk of hernia formation, strangulation, and high morbidity and mortality. Although laparoscopy has been proposed as a means of evaluating the diaphragm in these patients, prior studies did not include a confirmatory procedure or did not report long-term follow-up. Thus, true sensitivity and specificity remain unknown. The purpose of this study was to determine the sensitivity and specificity of laparoscopy for the detection of diaphragm injury after penetrating thoracoabdominal trauma. We hypothesized that laparoscopy alone is sufficient to exclude diaphragm injury after penetrating thoracoabdominal trauma. METHODS: We conducted a prospective case series of 34 hemodynamically normal asymptomatic patients with thoracoabdominal penetrating injuries. All patients underwent diagnostic laparoscopy to evaluate the diaphragm for the presence of injury. All patients then underwent confirmatory celiotomy (n = 30) or video-assisted thoracoscopy (n = 4). RESULTS: All patients were men between the ages of 18 and 54 years. There were 37 stab wounds and 1 gunshot wound. The mean lowest preoperative systolic blood pressure recorded was 120 +/- 18 mm Hg. Penetrating injuries were stratified by anatomic location (anterior, 18; posterior, 8; flank, 9; not specified, 3). There were 7 true-positive, 30 true-negative, no false-positive, and 1 false-negative result. Specificity, sensitivity, and negative predictive value were 100%, 87.5%, and 96.8%, respectively. The single missed injury occurred in a patient with hemoperitoneum from associated splenic injury that obscured the diaphragm and warranted celiotomy. CONCLUSION: In asymptomatic hemodynamically normal patients with penetrating thoracoabdominal injury, laparoscopy alone is sufficient to exclude diaphragmatic injury. PMID- 15824658 TI - Should patients with isolated hip fractures be included in trauma registries? AB - BACKGROUND: Patients with isolated hip fractures are frequently excluded from trauma registries. The goal of this study was to show that patients with these injuries have higher resource use and poorer outcomes than the rest of the trauma population. METHODS: The Quebec Trauma Registry was used to identify all trauma patients from April 1, 1998, to March 31, 2003. Patients who were dead on arrival at the emergency room were excluded. Isolated hip fracture (HIP) was defined as a diagnosis of a single fracture to the neck of the femur (Abbreviated Injury Scale 1990 codes 851808.3, 851810.3, 851812.3, and 851818.3) secondary to a fall and for which the Injury Severity Score was 9 or 10 (no other Abbreviated Injury Scale code higher than 1). Patients with all other trauma diagnosis (OT) were used for comparison. Outcome variables were length of hospital stay, length of intensive care unit (ICU) stay, in-hospital complications, and status and orientation at discharge. Chi-square and Wilcoxon rank-sum tests were used. RESULTS: There were 68,422 patients: 14,426 (21.1%) HIP patients and 53,996 (78.9%) OT patients. The median Injury Severity Score was 9 for HIP (range, 9-10) and 9 for OT (range, 1-75). Mean length of hospital stay was 18.4 days for HIP compared with 11.7 days for OT (p < 0.0001). HIP patients represented 29.5% of the total hospital stay. ICU stay was required for 1,353 HIP patients (9.4%) and for 12,395 (23.0%) OT patients (p < 0.0001). Mean ICU stay was 3.9 days for HIP compared with 5.5 days for OT (p = 0.0006). In-hospital mortality was 8.5% in HIP compared with 3.7% in OT (p < 0.0001). HIP represented 62.7% of patients referred for long-term care and 39.3% of patients referred to a rehabilitation center. CONCLUSION: Patients with HIP represented 21.1% of admissions while accounting for 42% of total days of hospitalization and 38% of deaths. Patients with hip fractures have a significantly higher risk of death, prolonged hospital stay, and complication rate, and are more often transferred to a rehabilitation center or to a long-term nursing home than the rest of the trauma population despite lower severity. They require multidisciplinary care typical of the rest of the trauma population and should be included in the trauma registry if the registry is to document the full outcome and resource use of the trauma population. PMID- 15824659 TI - Heart rate variability and spontaneous baroreflex sequences: implications for autonomic monitoring during hemorrhage. AB - BACKGROUND: Noninvasive procedures for predicting progression to hemodynamic instability during induced central hypovolemia in humans were evaluated. The purpose of this study was twofold: (1) to track changes in autonomic function induced by a model of hemorrhage, and (2) to determine whether measures of autonomic function are reliable without strict control of breathing. METHODS: Electrocardiogram, respiratory frequency, and arterial pressure during progressive lower body negative pressure were recorded for 10 subjects, and for a separate sample of 20 subjects during 5-minute periods of spontaneous breathing or controlled-frequency breathing at 15 breaths per minute.. Heart rate variability was calculated in both time and frequency domains. Up and down baroreflex sequences were calculated with linear regression analysis between sequential changes in systolic pressures and accompanying parallel changes in R-R intervals. RESULTS: Heart rate variability (r = 0.92) and up (r = 0.90) and down (r = 0.96) sequences changed in direct inverse relation to decreased central volume as produced by progressive increases in lower body negative pressure, whereas mean arterial pressures remained constant (r = 0.26). Neither heart rate variability nor up and down baroreflex sequences were affected by the mode of breathing. CONCLUSIONS: Analysis of heart rate variability and baroreflex sequences in hemorrhaging patients may provide advance recognition of those at risk for progression to shock. In conjunction with more traditional modes of assessing volume status, tracking early changes in autonomic function may improve resuscitation efforts for the hemodynamic compromised patient. PMID- 15824660 TI - Can near-infrared spectroscopy identify the severity of shock in trauma patients? AB - BACKGROUND: Our recent experimental study showed that peripheral muscle tissue oxygen saturation (StO2), determined noninvasively by near-infrared spectroscopy (NIRS), was more reliable than systemic hemodynamics or invasive oxygenation variables as an index of traumatic shock. The purpose of this study was to establish the normal range of thenar muscle StO2 in humans and the relationship between shock state and StO2 in trauma patients. METHODS: This was a prospective, nonrandomized, observational, descriptive study in normal human volunteers (n = 707) and patients admitted to the resuscitation area of our Level I trauma center (n = 150). To establish a normal StO2 range, an NIRS probe was applied to the thenar eminence of volunteers (normals). Subsequently, in a group of trauma patients, an NIRS probe was applied to the thenar eminence and data were collected and stored for offline analysis. StO2 monitoring was performed continuously and noninvasively, and values were recorded at 2-minute intervals. Five moribund trauma patients were excluded. Members of our trauma faculty, blinded to StO2 values, classified each patient into one of four groups (no shock, mild shock, moderate shock, and severe shock) using conventional physiologic parameters. RESULTS: Mean +/- SD thenar StO2 values for each group were as follows: normals, 87 +/- 6% (n = 707); no shock, 83 +/- 10% (n = 85); mild shock, 83 +/- 10% (n = 19); moderate shock, 80 +/- 12% (n = 14); and severe shock, 45 +/- 26% (n = 14). The thenar StO2 values clearly discriminated the normals or no shock patients and the patients with severe shock (p < 0.05). CONCLUSION: Decreased thenar muscle tissue oxygen saturation reflects the presence of severe hypoperfusion and near-infrared spectroscopy may be a novel method for rapidly and noninvasively assessing changes in tissue dysoxia. PMID- 15824661 TI - Increases in both buccal and sublingual partial pressure of carbon dioxide reflect decreases of tissue blood flows in a porcine model during hemorrhagic shock. AB - BACKGROUND: Earlier studies had demonstrated that gastric partial pressure of carbon dioxide (PCO2) of the gastric wall is an early indication of perfusion failure. Because hypercarbia is a general phenomenon of perfusion failure, this study investigated whether sublingual and buccal mucosal tissue PCO2 also serve as a sensitive indicator of systemic blood flow during hemorrhagic shock. METHODS AND RESULTS: Hemorrhagic shock was induced in five male domestic pigs. Buccal and sublingual PCO2 increased from 60 to 129 mm Hg (p < 0.01) in parallel with average decreases in cardiac output to 44% (from 6.1 to 2.8 L/minute; p < 0.01), and mean arterial pressure to 47% (from 115 to 57 mm Hg; p < 0.01), over the 2 hour interval of shock. According to colored microspheres used for measurements, buccal mucosal flow decreased to 35% (from 6.3 to 2.2 mL/minute/100 g; p < 0.01) in close parallel with sublingual blood flow, which decreased to 34% (from 7.2 to 2.5 mL/minute/100 g; p < 0.01). Liver flow decreased to 56% (from 152 to 85 mL/minute/100 g;p < 0.01), and renal flow to 47% (from 272 to 128 mL/minute/100 g;p < 0.01) of baseline values. The procedure yielded to overall comparable volume exchanges in different animals. After reinfusion of shed blood, buccal PCO2, like sublingual PCO2, was restored to approximately baseline values (respectively, from 56 to 71 mm Hg and from 60 to 71 mm Hg; p = not significant), together with arterial pressure, cardiac output, and end-tidal CO2 (EtCO2) (respectively, from 115 to 115 mm Hg, from 6.1 to 6.3 L/minute, and from 35 to 39 mm Hg; p = not significant), but there was delayed reversal of lactic acidosis (from 0.7 to 2.5 mmol/L;p < 0.01). In five unbled control animals, no significant changes were observed over the same interval. CONCLUSION: The current study extends the rationale for noninvasive measurements to both buccal and sublingual mucosa for diagnosis and quantitation of hemorrhagic shock severity. PMID- 15824662 TI - Early diagnosis of hypovolemic shock by sonographic measurement of inferior vena cava in trauma patients. AB - BACKGROUND: The diameter of the inferior vena cava in trauma patients may be useful for evaluating hypovolemia. METHODS: Between June 2003 and September 2003, 35 injured patients transferred to the authors' hospital were prospectively investigated. They were divided into two groups: a shock group (n = 10) and a control group (n = 25). The maximum anteroposterior diameter of the inferior vena cava was measured using a sonography at arrival and on hospital day 5. RESULTS: The average diameter of the inferior vena cava in the shock group was significantly smaller than in the control group. There was no significant change in the diameter of the inferior vena cava in the control group, but significant change was seen in the shock group between arrival and hospital day 5. CONCLUSION: The diameter of the inferior vena cava was found to correlate with hypovolemia in trauma patients. PMID- 15824663 TI - Validation of direct intraabdominal pressure measurement using a continuous indwelling compartment pressure monitor. AB - BACKGROUND: According to recommendations, intraabdominal pressure should be monitored every 8 hours for patients at high risk of abdominal compartment syndrome. Continuous intraabdominal pressure monitoring may be valuable for these patients. METHODS: For 15 patients undergoing laparoscopic surgery, a pressure monitor was introduced after formation of pneumoperitoneum. During the procedure, the laparoscopic insufflator pressure was varied. The pressure monitor values and the time to equilibrium were recorded. RESULTS: Altogether, 152 pressure recordings were taken for the patients studied. The measurements from the insufflator and pressure monitor were compared using a Bland-Altman plot. The mean difference between the techniques was 0.04 +/- 3.8, and 95% of the points from the pressure monitor were within two standard deviations of the mean difference. Pressure changes were essentially "real time." CONCLUSIONS: The intracompartmental pressure monitor provides accurate, rapid, and direct measurement of intraabdominal pressure, and may be a useful tool for continuous intraabdominal pressure measurement among patients at risk of abdominal compartment syndrome. PMID- 15824664 TI - Spine injuries are common among asymptomatic patients after gunshot wounds. AB - BACKGROUND: Spine injuries after gunshot wounds are thought to be rare among asymptomatic patients. The occurrence of spine injuries among asymptomatic patients with gunshot wounds was studied to determine the necessity for mandatory spine immobilization and radiographic imaging. METHODS: In this retrospective cohort study, initial physical examination, radiographic findings, and final diagnosis and treatment were reviewed. Patients were included if they were admitted to the authors' level 1 trauma center with gunshot wounds to the head, neck, or trunk during a 10-year period. Spine injuries were considered "significant" if the injury was associated with spinal cord injury or required spine-related surgical procedures or prolonged spine immobilization. Spine injuries were defined as "unsuspected" if there were no neurologic findings at admission. RESULTS: During the study period, 2,450 patients who survived more than 24 hours were admitted with gunshot wounds to the trunk, neck, or head. Of these patients, 244 (approximately 10%) had spine injuries, and 228 of them had complete records. Two thirds of the spine injuries were found to be significant, requiring surgery or prolonged immobilization, and 13% were unsuspected. CONCLUSIONS: Spine injuries without neurologic signs are not uncommon among patients with gunshot wounds. Complete radiographic spine imaging is therefore recommended to ensure that spine injuries are not missed in this population. PMID- 15824665 TI - Treatment of femoral shaft aseptic nonunion associated with broken distal locked screws and shortening. AB - BACKGROUND: This retrospective study investigated the treatment of femoral shaft aseptic nonunions associated with broken distal locked screws and shortening. METHODS: In this study, 11 femoral shaft aseptic nonunions associated with both broken distal locked screws and shortening in 11 consecutive adult patients were treated. All nonunions were associated with at least 1.5 cm (range, 1.5-3.5 cm) shortening. These nonunions were treated by removal of locked nails, one-stage femoral lengthening, static locked nail stabilization, and corticocancellous bone graft supplementation. Postoperatively, ambulation with protected weightbearing was encouraged as early as possible. RESULTS: Ten nonunions were followed up for a median of 4.1 years (range, 1.8-5.5 years), and nine fractures healed at a median of 4 months (range, 3-6 months). The nonunion case had broken locked screws again at 5 months and was treated with exchange nailing. The fracture healed uneventfully at 4 months. No other complications occurred. CONCLUSIONS: The key to removal of broken screws is withdrawal of the nail slightly to release the incarcerated broken screw end. The screw end then is pushed out with a used Knowles' pin or a smaller screwdriver under image intensifier guidance. Concomitant one-stage femoral lengthening to treat nonunion with shortening has a high success rate. PMID- 15824666 TI - Controlled study of in-line ovine spleen transection assisted by radiofrequency ablation. AB - BACKGROUND: Trauma to the spleen or tumors of the spleen often require total splenectomy for control of hemorrhage. Partial splenectomy is the preferred technique because of the short- and long-term sepsis problems in asplenic patients. Multiple techniques for partial splenectomy have been tried in the past with limited success. The authors designed the in-line radiofrequency ablation (ILRFA) probe for liver surgery. It uses radiofrequency energy to make a linear coagulative plane that allows the parenchyma of solid vascular organs to be divided. In this study, for the first time, the efficiency of ILRFA was tested with the ovine spleen. METHODS: Seven sheep were used for this study. With the sheep under general anesthesia, a laparotomy was performed. The first sheep was used for a pilot study. Eight partial splenectomies were made in the remaining six sheep using ILRFA. For a control, a matching partial splenectomy was made in each sheep using diathermy and sutures. Blood loss was measured by determining the difference in the weights of dry sponges and blood-stained sponges after resection. A paired t test was used to compare the bleeding between the control and the ILRFA techniques. RESULTS: The mean blood loss was 33.14 +/- 17 g using ILRFA and 123.43 +/- 72 g in the control group. The bleeding was significantly reduced in the ILRFA group (p = 0.0056). The time required to apply ILRFA was 12 minutes. CONCLUSION: Partial splenectomy was achieved in the ovine spleen using radiofrequency energy with minimal blood loss. PMID- 15824667 TI - Proximal left subclavian artery disruption caused by blunt trauma in an adolescent: case report and review of the literature. PMID- 15824668 TI - Liver transplantation in severe hepatic trauma after hepatic artery embolization. PMID- 15824669 TI - Extensive hydrofluoric acid injuries: a serious problem. PMID- 15824670 TI - Delayed penetrating intrathoracic injury from multiple rib fractures. PMID- 15824671 TI - Delayed bladder rupture after symphyseal hardware failure. PMID- 15824672 TI - Posterior sagittal anorectoplasty for the management of blunt anorectovaginal injury: case report. PMID- 15824673 TI - Novel therapies for sepsis: a review. AB - BACKGROUND: Over the past 30 years, efforts have been made to identify therapeutic targets in the host response to infection. METHODS: A review of the randomized controlled clinical sepsis trials and meta-analyses of glucocorticoids, mediator-specific anti-inflammatory agents, and anticoagulant agents was performed. RESULTS: The effects of glucocorticoids in sepsis appear to be dose-dependent, with high doses decreasing survival and low doses improving survival. As a class, the mediator-specific anti-inflammatory agents have a small beneficial effect on survival; however, no single agent has demonstrated significant benefit. The treatment effects of these agents appear to be related to the patient's risk of death. As a class, the anticoagulant agents do not improve survival; however, the efficacy of these agents may have been confounded by concurrent heparin therapy. Activated protein C demonstrated a beneficial effect on survival that was dependent on severity of illness. CONCLUSION: Trials of agents directed at altering the host's response during sepsis have had variable results, and it appears that several different factors may alter the efficacy of these agents. PMID- 15824674 TI - Traumatic pneumorrhachis. AB - Traumatic pneumorrhachis is extremely rare, and a review of the English literature has revealed only 31 reported cases. We review the cause and pathophysiology of this unique entity in trauma patients. We also discuss the significance of this entity to the trauma specialist, paying special attention to the differences between air in the subarachnoid space versus air in the epidural space. PMID- 15824675 TI - Bleeding post-traumatic pseudoaneurysm of the thigh. PMID- 15824676 TI - Civilian use of rotor wing medical transport has had its opponents. PMID- 15824677 TI - Cervical spine imaging in comatose patients. PMID- 15824680 TI - Twenty years of emergency medical services for children: a cause for celebration and a call for action. PMID- 15824681 TI - The use of high-dose epinephrine for patients with out-of-hospital cardiopulmonary arrest refractory to prehospital interventions. AB - OBJECTIVE: To determine if high-dose epinephrine (HDE) used during out-of hospital cardiopulmonary arrest refractory to prehospital interventions improves return of spontaneous circulation, 24-hour survival, discharge survival, and neurological outcomes. METHODS: A multicenter randomized controlled trial was conducted between May 1991 and October 1996 to compare the effectiveness of HDE versus standard-dose epinephrine (SDE) in patients having out-of-hospital cardiopulmonary arrest refractory to prehospital resuscitation efforts. Cardiopulmonary arrest was classified as "medical" or "traumatic." Two hundred thirty patients were enrolled in 7 pediatric emergency departments. Ages ranged from newborn to 22 years. Seventeen patients met exclusion criteria. Patients were assigned to receive HDE (0.1 mg/kg for the initial dose and 0.2 mg/kg for subsequent doses) or SDE (0.01 mg/kg). The main end points evaluated were return of spontaneous circulation, 24-hour survival, discharge survival, and neurological outcome. RESULTS: One hundred twenty-seven patients received HDE (32 trauma patients), and 86 patients received SDE (27 trauma patients). Among medical patients, 24 (25%) of 95 experienced return of spontaneous circulation in the HDE group as compared with 9 (15%) of 59 in the SDE group (P = 0.14, chi2 = 2.17, relative risk = 1.66 [0.83-3.31]). Sixteen (17%) of 95 HDE patients and 5 (8%) of 59 SDE patients survived at least 24 hours (P = 0.14, chi2 = 2.16, relative risk = 1.99 [0.77-5.14]). Nine survivors to discharge received HDE, and 2 received SDE (P = 0.21, Fisher exact test, relative risk = 2.75 [0.61-12.28]). There were no long-term survivors among the trauma patients. Eight of 11 long term survivors had severe neurological outcomes defined by the Glasgow Outcome Scale (2/2 SDE, 6/9 HDE; P = 0.51, Fisher exact test). CONCLUSION: HDE does not improve or diminish return of spontaneous circulation, 24-hour survival, long term survival, or neurological outcome compared with SDE in out-of-hospital cardiopulmonary arrest. PMID- 15824682 TI - Intervention to improve interobserver agreement in the assessment of children with pharyngitis. AB - OBJECTIVE: The objective of this study is to examine the effectiveness of an intervention to improve interobserver reliability of clinical evaluation of children with pharyngitis. METHODS: This was a sequential trial performed in an academic children's hospital emergency department. Two sets of 100 children aged 2 to 18 years with a complaint of sore throat were independently evaluated by emergency department attending staff and residents for coryza, scarlatiniform rash, tonsillar enlargement, tonsillar erythema, tonsillar exudates, cervical adenopathy, and palatal petechiae. The first 100 subjects were evaluated according to the clinician's routine practice; the second set was evaluated using a visual-aid sheet showing pictorial gradations of each characteristic. All findings were scored dichotomously and analyzed by kappa statistics. RESULTS: Both sets of patients were similar with respect to age, sex, race, and prior episodes of group A beta-hemolytic streptococcus. Interobserver reliability without the visual aid was poor in all areas except tonsillar enlargement and tonsillar exudates which had kappa values of 0.50 and 0.58, respectively. The use of the visual-aid sheet did not significantly improve reliability in any of the clinical evaluations, and all kappa scores remained below 0.50. CONCLUSION: Interobserver reliability in evaluating clinical signs of pharyngitis is fair to poor. The use of pictorial prompts does not improve interobserver reliability. PMID- 15824683 TI - Bronchiolitis in US emergency departments 1992 to 2000: epidemiology and practice variation. AB - OBJECTIVE: To describe the epidemiology of US emergency department (ED) visits for bronchiolitis, including the characteristics of children presenting to the ED and the variability in bronchiolitis care in the ED. METHODS: Data were obtained from the 1992 to 2000 National Hospital Ambulatory Medical Care Survey. Cases had International Classification of Diseases, Ninth Revision, Clinical Modification code 466 and were younger than 2 years. National estimates were obtained using assigned patient visit weights; 95% confidence intervals were calculated using the relative standard error of the estimate; analysis used chi2 and logistic regression. RESULTS: From 1992 to 2000, bronchiolitis accounted for approximately 1,868,000 ED visits for children younger than 2 years. Among this same age group, the overall rate was 26 (95% confidence interval 22-31) per 1000 US population and 31 (95% confidence interval 26-36) per 1000 ED visits. These rates were stable over the 9-year period. Comparing children with bronchiolitis to those presenting with other problems, children with bronchiolitis were more likely boys (61% vs. 53%; P = 0.01) and Hispanic (27% vs. 20%; P = 0.008). Therapeutic interventions varied and 19% were admitted to the hospital. The multivariate predictor for receiving systemic steroids was urgent/emergent status at triage (odds ratio 4.0, 1.9-8.4). Multivariate predictors for admission were Hispanic ethnicity (odds ratio 2.3, 1.1-5.0) and urgent/emergent status at triage (odds ratio 3.7, 2.0-6.9). CONCLUSIONS: ED visit rates for bronchiolitis among children younger than 2 years were stable between 1992 and 2000. The observed ED practice variation demonstrates that children are receiving medications for which there is little supporting evidence. Boys and Hispanics are at-risk groups for presentation to the ED, and Hispanics are more likely to be hospitalized. PMID- 15824684 TI - Unintentional childhood poisoning in the Sharon area in Israel: a prospective 5 year study. AB - OBJECTIVES: To study the epidemiology and risk factors for unintentional exposure to poisoning among the Jewish and the Arab population in the Sharon area in Israel. METHODS: We prospectively evaluated visits to the pediatric emergency department because of unintentional poisoning exposure, at the Meir General Hospital. We collected demographic data, substance exposure data, and the clinical outcome of the poisoning. RESULTS: During the 5 years of the study, 502 children were evaluated for unintentional poisoning, 84% Jewish and 16% Arabs; 88.5% occurred in children younger than 5 years, with a peak incidence at the age of 2 years (39.5%). Medications including hormones, vitamins, and antibiotics were the most common cause of exposure. Most children (95%) had no symptoms or abnormal findings on physical examination (84%), and most (85%) were discharged after several hours of observation. However, children of Arab origin presented with severe clinical manifestations because of a high rate of pesticide poisoning. There was 1 death from organic phosphate poisoning. CONCLUSION: Exposure to poisoning is commonly encountered in children. Pesticides exposure is more common in the Arab community and is usually associated with more severe clinical manifestations. Educational preventive programs are mandatory. PMID- 15824685 TI - Pediatric carbamazepine suspension overdose-clinical manifestations and toxicokinetics. AB - Two toddlers ingested unknown quantities of their older sibling's carbamazepine suspension and rapidly manifested central nervous depression requiring intubation in 1 patient. Coma was the primary clinical finding throughout their care with no anticholinergic syndrome, seizures, or dysrhythmia. Both patients recovered without sequelae within 24 hours. Initial carbamazepine concentrations were 36.6 and 22.7 mg/L. The elimination rates (zero-order kinetic) were approximately 1.4 and 0.75 mg/L per hour. We provide the first toxicokinetic data for carbamazepine suspension overdose in children. We confirm that the oral absorption of suspension carbamazepine is rapid necessitating prompt referral to a health care facility for this exposure. PMID- 15824686 TI - Two teenagers with syncope. AB - We report the cases of 2 teenagers with syncope and headaches who were subsequently found, on head computed tomography examinations, to have central nervous system etiologies (arteriovenous malformation and arachnoid cyst) of their syncope. These cases highlight the importance of a focused history and physical examination when evaluating patients who present with syncope. PMID- 15824687 TI - Sports-related pneumothorax in children. AB - Pneumothorax after blunt chest trauma in the absence of rib fractures is uncommon and has only rarely been reported as a result of sporting activity. Presentation may vary from an apparently normal physical examination in the presence of a small pneumothorax to hemodynamic compromise in the presence of a tension pneumothorax. High fitness levels in athletes may result in failure to recognize symptoms and delay diagnosis, potentially increasing morbidity. It is imperative for the emergency physician to exclude pneumothorax in children who present with chest pain after blunt chest trauma from sports injury. We report our experience with and the management of 3 patients with pneumothoraces. PMID- 15824688 TI - Ventriculoperitoneal shunt malfunction presenting with pleuritic chest pain. AB - BACKGROUND: Ventriculoperitoneal (VP) shunts are widely used for treating hydrocephalus. These devices are prone to malfunction with up to 70% requiring revision. Shunt infection and obstruction comprise the majority of malfunctions and usually present dramatically. However, rare presentations occur. METHODS/RESULTS: We report a rare case of VP shunt malfunction presenting with pleuritic chest pain. A 13-year-old girl with a VP shunt placed at birth for congenital hydrocephalus presented on multiple occasions with pleuritic chest pain, cough, and fever. She was diagnosed with an upper respiratory tract infection and discharged home. She returned with respiratory compromise, and chest x-ray depicted the shunt catheter in the pleural space with an associated pleural effusion and infiltrate. The patient fully recovered with intravenous antibiotics, thoracentesis, and placement of a new shunt system. CONCLUSIONS: VP shunt malfunction usually presents with signs and symptoms of increased intracranial pressure and/or infection. However, unusual presentations of malfunction may occur with signs and symptoms which appear unrelated to the shunt. Thus, all patients with VP shunts warrant a comprehensive evaluation. PMID- 15824689 TI - Nucleic acid amplification tests (polymerase chain reaction, ligase chain reaction) for the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae in pediatric emergency medicine. AB - Nucleic acid amplification tests, such as ligase chain reaction and polymerase chain reaction, offer potential advantages of speed, simplicity, and accuracy in the detection of genitourinary tract infection with Neisseria gonorrhoeae and Chlamydia trachomatis. Their appropriate use in pediatric emergency medicine depends on an understanding of their strengths and weaknesses. Problems arise in defining the sensitivity and, especially, specificity of these tests. The clinical scenario, the site of infection, the age and sex of the patient, and especially the presence or absence of medicolegal concerns strongly affect the applicability of these tests. The risk of false positives may be significant even when legal concerns do not arise and even if a highly specific test is used. This article reviews the uses and limitations of such tests in pediatric emergency medicine. Discussion is directed to both technical and practical considerations. PMID- 15824692 TI - ECGs in the ED. PMID- 15824693 TI - Swallowed Coke can tab: is it still stuck in the esophagus? PMID- 15824694 TI - Child passenger safety in the school-age population. PMID- 15824695 TI - Chilling lessons from the Shipman inquiry. PMID- 15824696 TI - Community matrons: what's in a name? PMID- 15824697 TI - Impact of a course on community nurses' practice: an evaluation. AB - An evaluation of a community course for nurses working in primary care was undertaken to ascertain whether it made an impact on the students' clinical practice. The course was undertaken by 16 nurses working within community nursing teams that included intermediate care, health visiting, school nursing and district nursing. Three quarters of the nurses had worked in the community for less than one year and half of these were newly qualified nurses. The evaluation consisted of semi-structured questionnaires, which were completed after the course and by the student and their mentor after 3 months. The assessment of practice document submitted by the students for assessment was also scrutinized for examples of changes students had made to their practice. The results demonstrate that the students felt that the community course made an impact on their practice and encouraged them to develop some of the key skills identified for community nurses. PMID- 15824698 TI - Irritable bowel syndrome: diagnosis and symptom management. AB - Irritable bowel syndrome (IBS) is a very common problem affecting 10% of the population at some time. Its cause and pathogenesis, however, remain poorly understood. Diagnosis is usually straightforward and detailed investigations are only required when the presentation is atypical. Treating IBS is always a challenge. The concerns and expectations of the patient must be met, along with any associated psychosocial issues. Medication on its own without addressing all of these issues is usually unsuccessful. PMID- 15824699 TI - Is tea tree oil effective at eradicating MRSA colonization? A review. AB - In vitro studies show that tea tree oil is capable of killing methicillin resistant Staphylococcus aureus (MRSA) in a laboratory setting. This review of randomized controlled trials (RCTs) was undertaken to find out whether it is effective at eradicating MRSA colonization compared to standard mupirocin-based regimens in colonized patients. A wide range of databases and internet sources were searched to identify published and unpublished studies. Two RCTs were found that researched the effectiveness of tea tree oil preparations against MRSA. One small RCT (n = 30) showed a large but non-significant improvement at eradicating MRSA compared to traditional treatment, whereas a larger study (n = 224) demonstrated little difference in rates of eradication overall (41% for tea tree and 49% for mupirocin, p = 0.286). However, the larger study found that those with nasal colonization receiving a tea tree regimen were more likely to remain colonized with MRSA in the nose (absolute risk increase 31%, p<0.001). Currently there is insufficient evidence to support the routine use of tea tree oil in clinical practice for eradication of MRSA colonization. PMID- 15824702 TI - Hypertension management: the primary care nursing role. AB - Hypertension is a prevalent chronic illness that is implicated in many cardiovascular diseases. Practice nurses and the district nursing team have a major role to play in its management. In this article, current British Hypertension Society guidelines are presented and discussed with regards to their evidence base. Implications for nursing practice are highlighted. PMID- 15824703 TI - Barbara Stuttle: a life-long champion of pioneering nurses. Interview by Tom Pollard. PMID- 15824704 TI - Communication skills and district nurses: examples in palliative care. AB - This article discusses some communication skills that may be challenging for district nurses who are working with palliative care patients. The article suggests that assessing what the patient knows about their illness is a key factor to effective communication. It pays particular attention to the first visit made by the district nurse. The article also discusses some specific situations that the district nurse may encounter, such as answering direct patient questions, denial, collusion and anger. A framework of communication styles developed by Wilkinson is used to understand the responses that may be given in such challenging situations. PMID- 15824705 TI - The law and intimate examinations and treatments. PMID- 15824706 TI - Will new matrons boost 'grey' health care? PMID- 15824707 TI - Zinc paste bandages and the treatment of Flegel's disease. AB - Such is their prevalence in the population that community nurses are very likely to encounter unfamiliar dermatological problems during their practice, many of which will have been self-treated for some time before professional help is sought. This article reports a case of a rare congenital condition called Flegel's disease, which leads to the presence of small horny papules on the skin, predominantly on the legs. In this case, the papules had deteriorated and were causing wider problems. PMID- 15824708 TI - Lymphoedema: recognition, assessment and management. AB - This article aims to review the function of the lymphatic system, what lymphoedema is and how this condition can be treated. It identifies the differences between oedemas with the emphasis on pure lymphoedema. Topics covered include differentiating between oedemas, basic treatment plans, the importance of early detection and appropriate referral to specialist practitioners. It highlights the need for all health professionals to have some understanding and awareness of this chronic condition. PMID- 15824709 TI - Assessment and management of fungating wounds: a review. AB - Fingating wounds always present a management challenge. Not only is it often difficult to manage the physical aspects of the wound - pain, bleeding, exudate and odour - but also the psychological impact of a fungating wound on the patient and their family or carers is often considerable. This review examines current practice in relation to fungating wounds, and emphasizes the central important of assessment, communication and sensitivity in nursing management. PMID- 15824710 TI - In an election year, focus on the positive. PMID- 15824711 TI - Clinical competence in sharp debridement: an innovative course. AB - A quick and effective way of promoting wound healing is to carry out sharp debridement where indicated, but this procedure carries a high level of clinical risk. Minimizing risk is an essential feature of clinical governance. A new course in wound debridement was developed at King's College, London in response to local NHS trust demand which required assessment of competence for this high risk procedure. This innovative development crossed traditional boundaries, and required high collaboration between education and practice. PMID- 15824716 TI - Losing and retrieving. PMID- 15824717 TI - Dementia treatment update. PMID- 15824718 TI - Relationship of dementia and body weight. PMID- 15824719 TI - Dementia: complex care needing ongoing assessment. PMID- 15824720 TI - Incorporating medication regimen reviews into the interdisciplinary care planning process. AB - Although many long-term care providers regard medication utilization reviews in terms of regulatory mandates, survey deficiencies, practice intrusion, and policy violations, not all consider these activities in such negative terms. This article describes the approach used by a large interdisciplinary team at a private continuing-care retirement community (CCRC) in North Carolina that includes medication regimen review in the larger context of resident care planning. PMID- 15824724 TI - Development of nurse competencies to improve dementia care. AB - The rapid increase in the number of elders who need dementia care and the critical need for skilled care providers prompted Florida legislators to enact legislation to improve the care of these residents. One component of the new legislation mandated dementia training for long-term care staff and led to the development of dementia care competencies that would guide a competency-based curriculum to meet the demand for training. The competencies, methods used for development, and information regarding how to access these newly developed resources are described in this article. PMID- 15824725 TI - Falls associated with dementia: how can you tell? AB - In any practice setting, professional nurses must be skilled to assess and manage medical problems rapidly. Post-fall assessment among those with dementia is a daunting task. Emergent medical problems after a fall are difficult to detect among those with dementia, because impaired communication portrays a false reality that the older adult is uninjured. Furthermore, early detection of potential post-fall injuries may not occur within a health care system that relies on verbal communication and eyewitness accounts as the main source of fall occurrence information. Professional nurses must turn to other means of fall detection. One such strategy relies on observable changes in function and behavior, such as a recent decline in mobility, change in gait, level of consciousness, neurological function or vital signs. Those most susceptible exhibit prior observable signs of visual-spatial impairment such as impaired clock drawing, gait apraxia, and inability to negotiate their environment without falling. PMID- 15824726 TI - Imposed versus involved: different strategies to effect driving cessation in cognitively impaired older adults. AB - In the lives of persons with cognitive impairments, a time comes when it is necessary to give up driving. This may be a particularly difficult issue for the driver, his or her family members, and their health care professionals. In this study, the phenomenon of unsafe driving by cognitively impaired older adults and the process of affecting driving cessation, was explored through guided interviews with 216 persons throughout the state of Florida. Participants included professionals working in the aging network, mobility counselors, safety officers, individuals with a mild degree of cognitive impairment and their family members. The strategies employed to influence or effect cessation could be categorized as those that involved the individual affected and those that were imposed on the individual. Conditions that appeared to influence whether driving cessation would occur were observed. There were distinct pros and cons to each strategy. In this article, specific information is provided to guide nurses working with cognitively impaired people who have become unsafe drivers. It is expected that driving cessation can be voluntarily effected in many cases but may have to be imposed in others. The skilled nurse will know when and which type of recommendation is appropriate. PMID- 15824727 TI - Subcortical vascular dementia. AB - Vascular dementia is the second most prevalent type of dementia in the United States today. This article includes a review of its pathophysiology, which involves the damage of small vessels in the brain, an abundance of which are in the subcortical region, thus creating a subcategory called subcortical vascular dementia (SVD). Various diseases, such as diabetes and high blood pressure, predispose the individual to damage to these small vessels. The symptoms of SVD are included as a review and helpful outline to differentiate SVD from Alzheimer's dementia and depression. Additionally, evidence-based interventions are reviewed. Nurses play a unique role in preventing and minimizing this dementia, which afflicts such a large percentage of our elderly population. PMID- 15824728 TI - The Frail Elderly Community-Based Case Management Project. AB - Nationally, the older population is expected to double in numbers through the year 2030. Health care providers are challenged to develop new models of care delivery for this unique population. Described in this article is one project that demonstrates successful outcomes for a community in eastern North Carolina. The model implemented community-based geriatric case management for frail elderly citizens residing in a private home or in an assisted living facility. Conventional hands-on delivery was combined with the distance-based convenience of telehealth. The outcomes prove this model to be cost-effective while improving quality of life for enrollees. PMID- 15824729 TI - Bik/NBK accumulation correlates with apoptosis-induction by bortezomib (PS-341, Velcade) and other proteasome inhibitors. AB - Proteasome inhibitors have emerged as promising anticancer therapeutic agents. Bortezomib (PS-341), a specific proteasome inhibitor, exhibits antitumor activity against a wide range of malignancies and has been approved by the US Food and Drug Administration for the treatment of relapsed or refractory multiple myeloma. However, the molecular mechanisms of bortezomib-mediated apoptosis remain unclear. To characterize the mechanisms of apoptosis induction by proteasome inhibitors, we examined levels of Bcl-2 protein family members (Bik/NBK, Bax, Bak, Bcl-2, and Bcl-XL), release of cytochrome c, and activation of caspase-9 and -3 in human colon cancer cell lines DLD1, LOVO, SW620, and HCT116; human lung cancer cell line H1299; and human ovarian cancer cell line SKOV3 after they were treated with bortezomib. The result showed that bortezomib induced rapid accumulation of Bik/NBK but not other Bcl-2 family members in all six cell lines. Bortezomib-mediated Bik/NBK accumulation and apoptosis were also observed in human embryonic kidney cells 293 and normal human bronchial epithelial cells. Moreover, dramatic Bik/NBK accumulation and apoptosis induction were observed when cells were treated with proteasome inhibitor MG132 and calpain inhibitor I (ALLN). Furthermore, no detectable changes in IkappaBalpha levels or in NFkappaB functionality were found after treatment with bortezomib. Finally, Bik/NBK accumulation was caused by stabilization of the protein from degradation and was associated with bortezomib cytotoxicity and apoptosis induction. Pretreatment of DLD1 cells with Bik/NBK siRNA reduced bortezomib-mediated Bik/NBK accumulation and cell death. Our results suggested that Bik/NBK is one of the mediators of proteasome inhibitor-induced apoptosis. PMID- 15824730 TI - Caspase-dependent processing activates the proapoptotic activity of deleted in breast cancer-1 during tumor necrosis factor-alpha-mediated death signaling. AB - Deleted in breast cancer-1 (DBC-1) was initially cloned from a homozygously deleted region in breast and other cancers on human chromosome 8p21, although no function is known for the protein product it encodes. We identified the generation of amino-terminally truncated versions of DBC-1 during tumor necrosis factor (TNF)-alpha-mediated apoptosis. Full-length 150 kDa DBC-1 underwent caspase-dependent processing during TNF-alpha-mediated death signaling, to produce p120 DBC-1 and p66 DBC-1 carboxy-terminal fragments. Endogenous DBC-1 localized to the nucleus in healthy cells, but localized to the cytoplasm during TNF-alpha-mediated apoptosis, consistent with the loss of the amino-terminus containing the nuclear localization signal. Overexpression of an amino-terminal truncated DBC-1, resembling p120 DBC-1, caused mitochondrial clustering, mitochondrial matrix condensation, and sensitized cells to TNF-alpha-mediated apoptosis. The carboxy-terminal coiled-coil domain of DBC-1 was responsible for the cytoplasmic and mitochondrial localization, and for the death-promoting activity of DBC-1. Thus, caspase-dependent processing of DBC-1 may act as a feed forward mechanism to promote apoptosis and possibly also tumor suppression. DBC 1, like its homolog cell cycle and apoptosis regulatory protein-1 (CARP-1), may function in the regulation of apoptosis. PMID- 15824731 TI - Impact of PKCdelta on estrogen receptor localization and activity in breast cancer cells. AB - Regulation of estrogen receptor (ER) function in breast cancer cells is a complex process involving different signalling mechanisms. One signal transduction component that appears to influence ER signalling is protein kinase C (PKC). PKCdelta is a particular isoenzyme of the novel PKC subfamily that plays a role in growth control, differentiation and apoptosis. The aim of the present study was to investigate the impact of PKCdelta on the regulation of the transcriptional activity of the human ERalpha. By using 12-O-tetradecanoylphorbol 13-acetate (TPA), Bryostatin1 and Rottlerin, we show that active PKCdelta is a proproliferative factor in estrogen-dependent breast cancer cells. Furthermore, activation of PKCdelta by TPA resulted in activation and nuclear translocation of ERalpha and in an increase of ER-dependent reporter gene expression. Transfection and expression of the regulatory domain RDdelta of PKCdelta, which is inhibitory to PKCdelta, inhibited the TPA-induced ERalpha activation and translocation. ERalpha was not phosphorylated by PKCdelta; however, glycogen synthase kinase-3 (GSK3) was identified as a substrate of PKCdelta. The expression of RDdelta resulted in a decrease of TPA-induced GSK3 phosphorylation and translocation into the nucleus. We suggest that GSK3 plays a role in the PKCdelta-related nuclear translocation of ERalpha. PMID- 15824732 TI - An essential role of Pak1 phosphorylation of SHARP in Notch signaling. AB - The p21-activated kinases (Paks), an evolutionarily conserved family of serine/threonine kinases, play an important role in cytoskeletal reorganization in mammalian cells. The Notch signaling pathway plays an important role in the determination of cell fate/differentiation in a number of organs. Notch signaling is a complex process, and the mechanism by which Notch regulates multiple cellular processes is intriguing. The expression of both Notch and Pak1 has been shown to be deregulated in several human cancers. Using yeast two-hybrid screening, we identified SHARP, one of the Notch signaling components, as a Pak1 interacting protein. We found that SHARP is a physiologic interacting substrate of Pak1, and that this interaction enhances SHARP-mediated repression of Notch target genes. Pak1 phosphorylation sites in SHARP were mapped to Ser3486 and Thr3568 within the SHARP repression domain. Mutation of Pak1 phosphorylation sites in SHARP, inhibition of Pak1 functions by a Pak1-autoinhibitory fragment (amino acids 83-149), or expression of Pak1-specific siRNA interfered with SHARP mediated repression of Notch target reporter gene activation. These results demonstrate that Pak1-SHARP interaction plays an essential role in enhancing the corepressor functions of SHARP, thereby modulating Notch signaling in human cancer cells. PMID- 15824733 TI - Genetic alterations of the KLF6 gene in gastric cancer. AB - The KLF6 is a zinc-finger tumor suppressor that is frequently mutated in several human cancers and broadly involved in differentiation and development, growth related signal transduction, cell proliferation, apoptosis, and angiogenesis. To determine whether genetic alterations of KLF6 gene are involved in the development and/or progression of gastric cancer, we have screened a set of 80 sporadic gastric cancers for mutations and allele loss of the KLF6 gene. Four missense mutations, S155R, P172 T, S180L, and R198 K, were detected in transactivation domain of the KLF6 gene and one of them had biallelic mutations with somatic mutation of one allele and loss of the remaining allele. All of the cases with mutation were of advanced intestinal-type gastric cancer with lymph node metastasis. In addition, 16 (43.2%) of 37 informative cases showed allelic loss at KLF6 locus. Interestingly, allelic loss was also frequent in intestinal type gastric cancer. Therefore, our data suggest that genetic alterations of KLF6 gene might play an important role in the development or progression of sporadic gastric cancers. PMID- 15824734 TI - Stress-specific signatures: expression profiling of p53 wild-type and -null human cells. AB - Gene expression responses of human cell lines exposed to a diverse set of stress agents were compared by cDNA microarray hybridization. The B-lymphoblastoid cell line TK6 (p53 wild-type) and its p53-null derivative, NH32, were treated in parallel to facilitate investigation of p53-dependent responses. RNA was extracted 4 h after the beginning of treatment when no notable decrease in cell viability was evident in the cultures. Gene expression signatures were defined that discriminated between four broad general mechanisms of stress agents: Non DNA-damaging stresses (heat shock, osmotic shock, and 12-O-tetradecanoylphorbol 13-acetate), agents causing mainly oxidative stress (arsenite and hydrogen peroxide), ionizing radiations (neutron and gamma-ray exposures), and other DNA damaging agents (ultraviolet radiation, methyl methanesulfonate, adriamycin, camptothecin, and cis-Platinum(II)diammine dichloride (cisplatin)). Within this data set, non-DNA-damaging stresses could be discriminated from all DNA-damaging stresses, and profiles for individual agents were also defined. While DNA damaging stresses showed a strong p53-dependent element in their responses, no discernible p53-dependent responses were triggered by the non-DNA-damaging stresses. A set of 16 genes did exhibit a robust p53-dependent pattern of induction in response to all nine DNA-damaging agents, however. PMID- 15824735 TI - Identification of novel VHL target genes and relationship to hypoxic response pathways. AB - Upregulation of hypoxia-inducible factors HIF-1 and HIF-2 is frequent in human cancers and may result from tissue hypoxia or genetic mechanisms, in particular the inactivation of the von Hippel-Lindau (VHL) tumour suppressor gene (TSG). Tumours with VHL inactivation are highly vascular, but it is unclear to what extent HIF-dependent and HIF-independent mechanisms account for pVHL tumour suppressor activity. As the identification of novel pVHL targets might provide insights into pVHL tumour suppressor activity, we performed gene expression microarray analysis in VHL-wild-type and VHL-null renal cell carcinoma (RCC) cell lines. We identified 30 differentially regulated pVHL targets (26 of which were 'novel') and the results of microarray analysis were confirmed in all 11 novel targets further analysed by real-time RT-PCR or Western blotting. Furthermore, nine of 11 targets were dysregulated in the majority of a series of primary clear cell RCC with VHL inactivation. Three of the nine targets had been identified previously as candidate TSGs (DOC-2/DAB2, CDKN1C and SPARC) and all were upregulated by wild-type pVHL. The significance for pVHL function of two further genes upregulated by wild-type pVHL was initially unclear, but re-expression of GNG4 (G protein gamma-4 subunit/guanine nucleotide-binding protein-4) and MLC2 (myosin light chain) in a RCC cell line suppressed tumour cell growth. pVHL regulation of CDKN1C, SPARC and GNG4 was not mimicked by hypoxia, whereas for six of 11 novel targets analysed (including DOC-2/DAB2 and MLC2) the effects of pVHL inactivation and hypoxia were similar. For GPR56 there was evidence of a tissue specific hypoxia response. Such a phenomenon might, in part, explain organ specific tumorigenesis in VHL disease. These provide insights into mechanisms of pVHL tumour suppressor function and identify novel hypoxia-responsive targets that might be implicated in tumorigenesis in both VHL disease and in other cancers with HIF upregulation. PMID- 15824736 TI - In Ewing's sarcoma CCN3(NOV) inhibits proliferation while promoting migration and invasion of the same cell type. AB - Altered expression of CCN3 has been observed in a variety of musculoskeletal tumours, including Ewing's sarcoma (ES). Despite its widespread distribution, very little is known about its biological functions and molecular mechanisms of action. We transfected CCN3 gene into a CCN3-negative ES cell line and analysed the in vitro and in vivo behaviours of stably transfected clones. Forced expression of CCN3 significantly reduced cell proliferation in vitro, growth in anchorage-independent conditions, and tumorigenicity in nude mice. Despite the antiproliferative effect, CCN3-transfected ES cells displayed increased migration and invasion of Matrigel. The decreased expression of alpha2beta1 integrin receptor and the increased amount of cell surface-associated matrix metalloproteinase (MMP)-9 following the expression of CCN3 may be the basis for the increased migratory abilities of transfected cells. Cells lacking alpha2beta1 are less facilitated to have stable anchorage since the predominant collagen extracted from ES tissue is indeed type I collagen, and proMMP-9 was recently found to provide a cellular switch between stationary and migratory ES cell phase. Our findings are in line with those recently obtained in glioblastoma. However, the underlying molecular mechanisms appear to be different, further highlighting the importance of the cellular context in the regulation of function of CCN proteins. PMID- 15824737 TI - Rare amplicons implicate frequent deregulation of cell fate specification pathways in oral squamous cell carcinoma. AB - Genomes of solid tumors are characterized by gains and losses of regions, which may contribute to tumorigenesis by altering gene expression. Often the aberrations are extensive, encompassing whole chromosome arms, which makes identification of candidate genes in these regions difficult. Here, we focused on narrow regions of gene amplification to facilitate identification of genetic pathways important in oral squamous cell carcinoma (SCC) development. We used array comparative genomic hybridization (array CGH) to define minimum common amplified regions and then used expression analysis to identify candidate driver genes in amplicons that spanned <3 Mb. We found genes involved in integrin signaling (TLN1), survival (YAP1, BIRC2), and adhesion and migration (TLN1, LAMA3, MMP7), as well as members of the hedgehog (GLI2) and notch (JAG1, RBPSUH, FJX1) pathways to be amplified and overexpressed. Deregulation of these and other members of the hedgehog and notch pathways (HHIP, SMO, DLL1, NOTCH4) implicates deregulation of developmental and differentiation pathways, cell fate misspecification, in oral SCC development. PMID- 15824738 TI - Recurrent chromosomal aberrations in INK4a/ARF defective primary lymphomas predict drug responses in vivo. AB - Predicting responsiveness to anticancer therapy based on molecular findings at diagnosis is important to optimize treatment decisions. Although clinical outcome correlates with distinct mutations in some cancer entities, treatment responses within these lesion-stratified subgroups still remain heterogeneous, underscoring the need for additional prognosticators. We previously demonstrated that defined genetic defects at the INK4a/ARF locus, which encodes the tumor suppressors p16INK4a and ARF, not only accelerated lymphomagenesis in the Emu-myc transgenic mouse but also interfered with treatment sensitivity. In this study, we take a nonbiased genome-wide approach to examine whether the responsiveness of these lymphomas can be further stratified based on cytogenetic information at diagnosis. Indeed, using spectral karyotyping, comparative genomic hybridization, and fluorescence in situ hybridization in 38 primary lymphomas, we find recurrent cytogenetic alterations that refine the predictive value of INK4a/ARF lesions on drug responses in vivo: gain of chromosome 14, which was never detected in INK4a/ARFnull lymphomas, defined an ARFnull subgroup with superior treatment outcome. Gain of chromosome 6 was identified as a recurrent chromosomal aberration that predisposed ARFnull tumors to their subsequent INK4a loss during therapy. These data illustrate how cytogenetic information from cancer specimens might complement established prognostic markers and may improve anticancer treatment strategies. PMID- 15824739 TI - Inactivation of p16, RUNX3, and HPP1 occurs early in Barrett's-associated neoplastic progression and predicts progression risk. AB - Patients with Barrett's esophagus (BE) are at increased risk of developing esophageal adenocarcinoma (EAC). Clinical neoplastic progression risk factors, such as age and the length of the esophageal BE segment, have been identified. However, improved molecular biomarkers predicting increased progression risk are needed for improved risk assessment and stratification. Using real-time quantitative methylation-specific PCR, we screened 10 genes (HPP1, RUNX3, RIZ1, CRBP1, 3-OST-2, APC, TIMP3, p16, MGMT, p14) for promoter hypermethylation in 77 EAC, 93 BE, and 64 normal esophagus (NE) specimens. A subset of genes manifesting significant differences in methylation frequencies between BE and EAC was then analysed in 20 dysplastic specimens. All 10 genes except p14 were frequently methylated in EACs, with RUNX3, HPP1, CRBP1, RIZ1, and OST-2 representing novel methylation targets in EAC and/or BE. p16, RUNX3, and HPP1 displayed increasing methylation frequencies in BE vs EAC. Furthermore, these increases in methylation occurred early, at the interface between BE and low-grade dysplasia (LGD). To demonstrate the silencing effect of hypermethylation, we selected the EAC cells BIC1, in which the HPP1 promoter is natively methylated, and subjected them to 5 aza-2'-deoxycytidine (Aza-C) treatment. Real-time RT-PCR indicated increased HPP1 mRNA levels after 3 days of Aza-C treatment, as well as decreased levels of methylated HPP1 DNA. Hypermethylation of a subset of six genes (APC, TIMP3, CRBP1, p16, RUNX3, and HPP1) was then tested in a retrospective longitudinal study of 99 BE and nine LGD specimens obtained from 53 BE patients undergoing surveillance endoscopy. Only high-grade dysplasia (HGD) or EAC were defined as progression end points. Two patient groups were compared: eight progressors (P) and 45 nonprogressors (NP), using Cox proportional hazards models to determine the relative progression risks of age, BE segment length, and methylation events. Multivariate analyses revealed that only hypermethylation of p16 (odds ratio (OR) 1.74, 95% confidence interval (CI) 1.33-2.20), RUNX3 (OR 1.80, 95% CI 1.08-2.81), and HPP1 (OR 1.77, 95% CI 1.06-2.81) were independently associated with an increased risk of progression, whereas age, BE segment length, and hypermethylation of TIMP3, APC, or CRBP1 were not independent risk factors. In combined analyses, risk was detectable up to, but not earlier than, 2 years preceding neoplastic progression. Hypermethylation of p16, RUNX3, and HPP1 in BE or LGD may represent independent risk factors for the progression of BE to HGD or EAC. These findings have implications regarding risk stratification, early EAC detection, and the appropriate endoscopic surveillance interval for patients with BE. PMID- 15824740 TI - Estrogen receptor positivity in mammary tumors of Wnt-1 transgenic mice is influenced by collaborating oncogenic mutations. AB - The majority (75%) of human breast cancers express estrogen receptor (ER). Although ER-positive tumors usually respond to antiestrogen therapies, 30% of them do not. It is not known what controls the ER status of breast cancers or their responsiveness to antihormone interventions. In this report, we document that transgenic (TG) expression of Wnt-1 in mice induces ER-positive tumors. Loss of Pten or gain of Ras mutations during the evolution of tumors in Wnt-1 TG mice has no effect on the expression of ER, but overexpression of Neu or loss of p53 leads to ER-negative tumors. Thus, our results provide compelling evidence that expression of ER in breast cancer may be influenced by specific genetic changes that promote cancer progression. These findings constitute a first step to explore the molecular mechanisms leading to ER-positive or ER-negative mammary tumors. In addition, we find that ER-positive tumors arising in Wnt-1 TG mice are refractory to both ovariectomy and the ER antagonist tamoxifen, but lose ER expression with tamoxifen, suggesting that antiestrogen selects for ER-negative tumor cells and that the ER-positive cell fraction is dispensable for growth of these tumors. This is a first report of a mouse model of antiestrogen-resistant ER-positive breast cancers, and could provide a powerful tool to study the molecular mechanisms that control antiestrogen resistance. PMID- 15824741 TI - A mutation-created novel intra-exonic pre-mRNA splice site causes constitutive activation of KIT in human gastrointestinal stromal tumors. AB - We report a new mechanism of aberrant pre-mRNA splicing resulting in constitutive activation of a mis-spliced oncoprotein (KIT) leading to malignancy (gastrointestinal stromal tumor) in contrast to loss of function of mis-spliced proteins resulting in diverse human diseases in the literature. The mechanisms of three consecutive molecular events, deletion of noncoding and coding regions encompassing the 3' authentic splice site, creation of a novel intra-exonic pre mRNA 3' splice acceptor site leading to in-frame loss of 27 nucleotides (nine amino acids; Lys550-Lys558), and the mechanism of constitutive activation of the mis-spliced KIT are elucidated. Loss of a peptide in a critical location unleashed the protein from autoinhibition (as evidenced by three-dimensional structural analysis), causing KIT to become constitutively activated and resulting in the GIST phenotype. We also demonstrated that only one of the following two exonic splicing enhancers is sufficient for inclusion of the KIT exon 11 in vivo: AACCCATGT (nucleotides 2-10 from the 5' end, which are recognized by SC35, SRp55, and SF2/ASF) or GGTTGTTGAGG (nucleotides 27-37 from the 5' end, which are recognized by SC35 and SRp55), suggestive of exonic enhancer redundancy. PMID- 15824742 TI - Roles of thioredoxin reductase 1 and APE/Ref-1 in the control of basal p53 stability and activity. AB - The p53 protein is redox-sensitive in vitro but in vivo effectors of this sensitivity are not known. In yeasts deficient for thioredoxin (Trx) reductase (TRR), p53 accumulates in an inactive, oxidized form, suggesting a role for TRR Trx in controlling p53. In mammalian cells, p53 binds to redox factor-1 (APE/Ref 1), an enzyme containing an abasic endonuclease domain involved in base excision repair, and a thiol reductase domain recycled by Trx and involved in regulating the transcription factor AP-1. To evaluate the role of TRR and APE/Ref-1 in p53 regulation, we have abrogated their expression using RNA interference in cell lines expressing wild-type p53. Inhibition of TRR resulted in accumulation of oxidized Trx and increased levels and DNA-binding activity of p53, with no phosphorylation of Ser15 or Ser20. In contrast, inhibition of APE/Ref-1 accelerated p53 protein turnover, resulting in a decrease in p53 levels and activity. However, inhibition of either TRR or APE/Ref-1 did not prevent activation and accumulation of p53 in response to DNA-damage by doxorubicin. When both factors were inhibited, basal levels of p53 were restored. These results suggest that TRR-Trx and APE/Ref-1 cooperate in the control of basal p53 activity, but not in its induction by DNA-damage. PMID- 15824743 TI - Quantitative genome scan and Ordered-Subsets Analysis of autism endophenotypes support language QTLs. AB - Autism is a neurodevelopmental syndrome with early childhood onset and deficits in three behavioral and cognitive dimensions: language, social skills and repetitive or restrictive behaviors. We hypothesized that using these endophenotypes would provide more power to detect linkage than the diagnosis of autism. Previously, we reported results for a nonparametric quantitative trait locus (QTL) genome scan in 152 families with autism, which revealed a linkage peak related to spoken language on 7q35. Here, we present the results of a nonparametric QTL scan of autism endophenotypes in 291 multiplex families, including the original 152. The strongest evidence for an 'age at first word' QTL was on chromosomes 3q at 147 cM (Z=3.10, P<0.001), and 17q at 93 cM (Z=2.84, P=0.002), both represent novel susceptibility loci for autism endophenotypes. There was also support for a previously identified autism peak on chromosome 17 at 43 cM (Z=2.22, P=0.013) with 'age at first phrase'. The 7q35 language peak was attenuated (Z=2.05, P=0.02) compared with the original finding. To explore the possibility of increased heterogeneity resulting from the addition of 135 families to the sample, we conducted an Ordered-Subsets Analysis on chromosome 7; these results suggest that the 132 autism families with the earliest average age at first word are responsible for the QTL on 7q35. This locus on 7q35 may harbor a gene contributing variability in spoken language that is not uniquely related to language delay in autism. PMID- 15824744 TI - Lack of association of the COMT (Val158/108 Met) gene and schizophrenia: a meta analysis of case-control studies. AB - There is strong evidence for a genetic contribution to schizophrenia, but the contribution of individual candidate genes remains uncertain. We attempted to replicate a recent meta-analysis that reported an association of the catechol O methyltransferase (COMT) Val allele with schizophrenia, and suggested that this effect may be moderated by ancestry. We included reports published subsequent to the original meta-analysis, and included a formal test of the moderating effect of ancestry in order to test whether the association operates differently in populations of European ancestry compared to populations of Asian ancestry. A corrected P-value for the 5% significance threshold was employed where appropriate, using Bonferroni's method, and studies that demonstrated departure from Hardy-Weinberg equilibrium among controls were excluded. When all studies were included in a meta-regression, there was evidence for a significant association of COMT Val allele frequency with schizophrenia case status and a significant main effect of ancestry. The interaction of COMT Val allele frequency and ancestry was also significant. However, when only studies that reported allele frequencies that did not depart significantly from Hardy-Weinberg equilibrium among controls were included, these effects were no longer significant. The results of our meta-analysis do not support an association between the COMT Val allele and schizophrenia case status, and do not support recent claims that this association may be moderated by ancestry. PMID- 15824745 TI - Population-based and family-based studies on the serotonin transporter gene polymorphisms and bipolar disorder: a systematic review and meta-analysis. AB - The serotonin transporter (5-HTT) is a candidate gene for bipolar disorder (BPD). It has been investigated for association with the illness in a series of studies, but overall results have been inconsistent and its role in the disorder remains controversial. Systematic reviews using meta-analytical techniques are a useful method for objectively and reproducibly assessing individual studies and generating combined results. We performed two meta-analyses of published studies- both population-based and family-based studies--investigating the association between BPD and the 5-HTT gene-linked polymorphic region (5-HTTLPR) and the intron 2 variable numbers of tandem repeats (VNTR) polymorphisms. The literature was searched using Medline and Embase to identify studies for inclusion. We statistically joined population-based and family-based studies into a single meta analysis. For both polymorphisms, our review revealed significant pooled odds ratios (ORs): 1.12 (95% CI 1.03-1.21) for the 5-HTTLPR and 1.12 (95% CI 1.02 1.22) for the intron 2 VNTR. Meta-regression showed that neither the study type (population-based vs family-based; P=0.41 for the 5-HTTLPR and P=0.91 for the intron 2 VNTR) nor the sample ethnicity (Caucasian vs non-Caucasian; P=0.35 for the 5-HTTLPR and P=0.66 for the intron 2 VNTR) significantly contributed to the heterogeneity of the meta-analyses. The observed ORs could be regarded simply as a very small but detectable effect of the 5-HTT, which has an additive effect when combined with other susceptibility loci. Alternative hypotheses on this finding were also discussed: a stronger effect of the haplotypes involving the two polymorphisms or other SNP markers; a more direct effect of these polymorphisms on specific phenotypes of BPD; and the presence of gene-environment interaction as a mediator of the genetic effects of 5-HTT. PMID- 15824746 TI - Aerobic exercise, lipids and lipoproteins in overweight and obese adults: a meta analysis of randomized controlled trials. AB - OBJECTIVE: Use the meta-analytic approach to examine the effects of aerobic exercise on lipids and lipoproteins in overweight and obese adults. DATA SOURCES: (1) Computerized literature searches, (2) cross-referencing from review and original articles, (3) hand searching, and (4) expert review of reference list. STUDY SELECTION: (1) randomized controlled trials, (2) aerobic exercise > or =8 weeks, (3) adult humans > or =18 y of age, (4) all subjects overweight or obese (BMI > or =25 kg/m(2)), (5) studies published in journal, dissertation, or master's thesis format, (6) studies published in the English-language, (7) studies published between 1 January 1955 and 1 January 2003, (8) assessment of one or more of the following lipid and/or lipoprotein variables: total cholesterol (TC), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), and triglycerides (TG). DATA ABSTRACTION: Dual coding by the first two authors (inter-rater agreement=0.96). RESULTS: In total, 13 studies representing 31 groups (17 exercise, 14 control), 613 subjects (348 exercise, 265 control), and up to 17 outcomes were available for pooling. Across all categories, random-effects modeling resulted in statistically significant improvements for TC (X +/- s.e.m., - 3.4+/-1.7 mg/dl, 95% CI, - 6.7 to - 0.2 mg/dl) and TG (X +/-s.e.m., - 16.1+/-7.3 mg/dl, 95% CI, - 30.2 to - 2.1 mg/dl) but not HDL (X +/- s.e.m., 1.6+/-0.8 mg/dl, 95% CI, - 0.02 to 3.2 mg/dl) or LDL (X +/-s.e.m., - 0.5+/-1.3 mg/dl, 95% CI, - 3.0 to 2.0 mg/dl). Changes were equivalent to improvements of 2% (TC), 11% (TG), 3% (HDL), and 0.3% (LDL). After conducting sensitivity analyses (each study deleted from the model once), only decreases in TG remained statistically significant. Increases in HDL were associated with increases in maximum oxygen consumption (VO(2 max) in ml/kg/min, r=0.75, P=0.002) and decreases in body weight (r=0.77, P<0.001), while decreases in LDL were associated with decreases in body weight (r=0.75, P=0.009). CONCLUSIONS: Aerobic exercise decreases TG in overweight and obese adults. However, a need exists for additional randomized controlled trials in various overweight and/or obese populations above and beyond those included in our analysis. PMID- 15824747 TI - Is there a causal relation between obesity and asthma? Evidence from Chile. AB - OBJECTIVE: An association between obesity and asthma symptoms has been reported in the literature, but such a relationship is inconsistent if atopic status or bronchial hyper responsiveness (BHR) is considered. The objective was to assess the association between obesity and asthma symptoms or BHR in adults. METHODS: A study was carried out in 1232 people born between 1974 and 1978 in Chile. The participants completed the European Community Health Survey questionnaire, were skin tested and subject to a BHR challenge to methacholine. MEASUREMENTS: Weight, height and waist circumference were measured and body mass index (BMI) was calculated. RESULTS: There was a positive association between wheeze and breathlessness following exercise and BMI (both with an OR 1.03, 95% CI 1.00 1.06), the associations with wheeze tended to disappear in women who did react at least to one allergen, and persisted in those who did not react to any allergens. BMI was negatively associated with BHR (OR 0.93, 95% CI 0.89-0.97). Waist circumference was not associated with asthma symptoms and it was negatively associated with BHR. CONCLUSION: Although there was an association between BMI and asthma symptoms, there were weaknesses in the evidence because waist circumference, a more direct measure of obesity than BMI, was not associated with asthma symptoms, and BMI and waist circumference were negatively associated with BHR. PMID- 15824749 TI - Body fat measurement in Indian men: comparison of three methods based on a two compartment model. AB - Obesity is a major risk factor for diabetes and related disorders. The current classification of obesity is based on body mass index (BMI, kg/m(2)), which is a surrogate for the total body fat. Since the relationship between BMI and body fat varies in different populations, an independent validation of the BMI-body fat relationship in the population of interest is desirable. OBJECTIVES: (1) To study the validity of field methods of measuring body fat (multiple skinfolds and bioimpedance) against a criterion method (deuterium dilution) and (2) To compare the prevalence of obesity (WHO 2000 criteria for BMI) with adiposity (body fat >25%) in middle-aged Indian men in rural and urban Pune. DESIGN: Community-based multistage stratified random sampling of middle-aged men from rural and urban Pune for study of body composition and cardiovascular risk. A third of these men, selected to represent wide BMI distribution, were studied for body fat measurements by specific methods. SUBJECTS: A total of 141 healthy men, approximately similar number from rural, urban slums and middle class from Pune. They were 39.3 (+/-6.2) y old and had a BMI of 21.9 (+/-3.7) kg/m(2). MEASUREMENTS: Anthropometry (height, weight and multiple skinfold thicknesses) by trained observers using standardised technique to calculate body fat by Durnin and Womersley's equation. Total body water and body fat by bioelectrical impedance analysis (BIA) and deuterium oxide dilution (D(2)O). RESULTS: Mean total body fat was 14.3 kg (23.0%) by anthropometry, 16.5 kg (26.0%) by BIA and 15.3 kg (24.6%) by D(2)O method. Although there was a good correlation between fat estimation by three methods (r= approximately 0.9, P<0.001 all), compared to D(2)O method anthropometry underestimated body fat by 1.0 kg and BIA overestimated fat by 1.2 kg (P<0.001 both). Using the standard cut-point of 25% body fat for 'adiposity' 29.5% rural, 46.0% slum and 75.0% middle class men were adipose. These proportions were considerably higher than the number of men who were 'preobese' (BMI> or =25-29.9 kg/m(2), 9.0% rural, 22.0% urban slums and 27.0% urban middle class) and 'obese' (BMI >30 kg/m(2), 4.0% urban slums, none in rural and urban middle class). CONCLUSION: We recommend that future studies assessing risk for chronic diseases in Indians should measure adiposity by anthropometry (multiple skinfolds) or BIA (calibrated for Indians) rather than relying only on BMI cut-points. PMID- 15824748 TI - Adiponectin receptor gene expression in human skeletal muscle cells is not regulated by fibrates and thiazolidinediones. AB - BACKGROUND: Thiazolidinediones as PPARgamma agonists and fibrates as PPARalpha agonists improve insulin sensitivity in insulin-responsive tissues. Recent data show an induction of adiponectin receptor 2 (AdipoR2) by PPARalpha and PPARgamma agonists in human macrophages. OBJECTIVE: In this study, we examined the effects of thiazolidinediones and fibrates on the expression of adiponectin receptors in human skeletal muscle cells, an important cell type in the context of insulin resistance. RESULTS AND METHODS: In vitro differentiated human myotubes treated with troglitazone or rosiglitazone (20 h) showed no significant changes in AdipoR1 and AdipoR2 mRNA expression. PPARgamma activation was controlled by determination of PPARgamma mRNA induction. Likewise, differentiated myotubes treated with Wy-14,643 or fenofibrate (20 h) revealed no significant regulation of AdipoR1 and AdipoR2 mRNA. PPARalpha activation was assessed by measuring PDHK4 mRNA expression. CONCLUSION: Induction of AdipoR gene expression in human skeletal muscle cells is not involved in the insulin-sensitizing effects of thiazolidinediones or fibrates. PMID- 15824751 TI - The Third International Symposium on Obesity and Hypertension ISOH'03: 'Genetics and Molecular Mechanisms' (October 23-25, 2003, Berlin Germany). AB - The Third International Symposium on Obesity and Hypertension (ISOH'03) was held on 23-25, October 2003 at the Max Delbruck Center for Molecular Medicine in Berlin-Buch, Germany. The meeting, which consisted largely of invited lectures, presented a state-of-the-art overview of the genetic and molecular mechanisms that link obesity and hypertension. The over 40 oral presentations were supplemented by around 90 poster presentations from 34 countries. The meeting was attended by around 250 participants from 54 countries. This paper briefly reviews the contents of the invited lectures presented at this meeting covering topics ranging from genetics, molecular mechanisms, pathophysiology, cardiovascular risk, to the management of patients with obesity-related hypertension. Stimulated by the continuing success of these Symposia, the organizers are currently planning to hold a Fourth International Symposium on Obesity and Hypertension (ISOH'05) in Berlin: a tentative date for this meeting has been set for 27-29, October 2005. PMID- 15824750 TI - Inhibition of appetite by nasal leptin administration in rats. AB - OBJECTIVE: Leptin inhibits appetite and reduces body weight. However, subcutaneous leptin administration is not very effective on weight reduction. The present studies were undertaken to test the hypotheses that nasally administered leptin effectively accesses to the brain and inhibits appetite. METHODS: Recombinant leptin (0.5 mg/rat) was administered into the bilateral nasal spaces of rats (i.n.). Changes in serum immunoreactive leptin (IRL) and cerebrospinal fluid (CSF)-IRL concentrations after i.n. leptin administration were compared after intraperitoneal (i.p.) administration. The influence of 0.1 or 0.5% lysophosphatidylcholine (LPC) as an optimizer of leptin absorption was examined. The anorexic effects of i.n. leptin were compared with i.p. leptin in ad libitum fed rats. RESULTS: The i.n. leptin increased CSF-IRL concentrations, although serum IRL concentrations of rats administered leptin i.n. were lower than those administered i.p. The addition of 0.1 and 0.5% LPC dose-dependently increased serum IRL concentrations, but did not modify CSF-IRL concentrations in i.n. leptin-treated rats. The i.n. leptin inhibited dark-time food consumption at 0-1 h and 3-6 h in ad libitum fed rats. In contrast, i.p. leptin reduced food consumption only for an hour. Phosphorylated signal transducer and activator of transcription (STAT) 3 immunoreactive cells increased in the arcuate nucleus (ARC) of the hypothalamus at 3 h only following i.n. leptin. CONCLUSION: The present study demonstrated that i.n. leptin caused longer inhibition of appetite and phosphorylation of STAT3 in ARC. It is concluded that the trans-nasal route may be useful for the selective access of leptin to the brain in obese people. PMID- 15824752 TI - Lower-body fat mass as an independent marker of insulin sensitivity--the role of adiponectin. AB - AIMS: To study the association between lower-body fat and estimates of whole-body insulin sensitivity in middle-aged men with and without a history of juvenile onset obesity, and to determine the possible mediating role of fasting serum adiponectin level as an insulin-sensitizing peptide. METHODS: A total of 401 men aged 39-65 y, body mass index 18-54 kg/m2, participated in the study. The following variables were measured on the study participants: regional body fat distribution as assessed by dual energy X-ray absorptiometry, abdominal sagittal diameter, maximal oxygen uptake (VO2max), physical activity, fasting and post glucose load levels of plasma glucose, serum insulin, and blood non-esterified fatty acid plus fasting levels of serum adiponectin and HbA1c. RESULTS: Lower body fat mass was positively associated with insulin sensitivity as estimated by Matsudas index also after adjusting for age, lean tissue mass, trunkal fat mass, weight changes since draft board examination, VO2max and the level of physical activity. In a subgroup of men selected for a large lower-body fat mass, fasting serum insulin concentration was 24% lower (P<0.01) and fasting serum adiponectin 33% higher (P<0.005) compared to a subgroup of men with a small lower-body fat mass but with similar trunkal fat mass. CONCLUSION: Lower-body fat mass is positively associated with an estimate of insulin sensitivity independently of trunkal fat mass in both lean and obese middle-aged men and this effect could partly be statistically explained by variations in serum adiponectin levels. PMID- 15824753 TI - Influence of CYP2C9 polymorphisms, demographic factors and concomitant drug therapy on warfarin metabolism and maintenance dose. AB - Warfarin is an anticoagulant drug with narrow therapeutic index and high interindividual variability in dose requirement. S-warfarin is metabolized mainly by polymorphic cytochrome P450 (CYP) 2C9. We systematically quantified the influence of CYP2C9 genotype, demographic factors and concomitant drug treatment on warfarin metabolism and maintenance dose. The mean warfarin doses were lower in carriers of one (2.71 mg/day, 59 patients) and two polymorphic alleles (1.64 mg/day, 11 patients) than in carriers of two wild-type alleles (4.88 mg/day, 118 patients). Multiple regression analysis demonstrated that CYP2C9 genotype, age, concomitant treatment with warfarin metabolism inducers and lean body weight contributed significantly to interindividual variability in warfarin dose requirement (adjusted R(2)=0.37). The same factors, except for age, significantly influenced S-warfarin clearance (adjusted R(2)=0.42). These results can serve as a starting point for designing prospective studies in patients in the initiation phase of genotype-based warfarin therapy. PMID- 15824754 TI - Nontraumatic spinal cord lesions: a prospective study of medical complications during in-patient rehabilitation. AB - STUDY DESIGN: Prospective study between 1st Jan 1995 and 31st Dec 1999. OBJECTIVE: To document medical complications among subjects with Nontraumatic Spinal Cord Lesions (NTSCL) during in-patient rehabilitation. SETTING: Bangalore, India. METHODS: Persons with NTSCL admitted for in-patient rehabilitation were included in the study. Clinical evaluation was carried out according to The International Standards for Neurological and Functional classification of Spinal Cord Injury. Disability was quantified using Barthel index. All medical complications were documented. RESULTS: A total of 297 subjects (154 men and 143 women) with NTSCL were included. The number of medical complications in each patient varied from 0 to 17 (mean=6.1+/-3.7). Common complications seen were urinary tract infections (184), spasticity (169), pain (149), urinary incontinence (147), depression (114), respiratory tract infections (101), constipation (92), pressure ulcers (89), contractures (52) and sleep disturbance (43). The number of medical complications correlated positively with duration of stay (Pearson's correlation coefficient r=0.5, P<0.01) and negatively with Barthel Index at admission (r=-0.2, P<0.05) and at discharge (r=-0.2, P<0.05). Complications were more frequent among people with tetraplegia than those with paraplegia (P<0.001). CONCLUSIONS: Medical complications are frequent among subjects undergoing rehabilitation for NTSCL. Patients with severe disability at admission have more complications during rehabilitation. Conversely, individuals with more complications have greater disability at discharge. PMID- 15824755 TI - The effect of passive cycling movements on spasticity after spinal cord injury: preliminary results. AB - OBJECTIVE: To investigate the influence of rhythmic passive movements of the legs on the reduction of spasticity after spinal cord injury (SCI). SETTING: Swiss Paraplegic Centre Nottwil, Switzerland. METHODS: A total of 10 subjects with motor complete SCI were treated with a cycling device for half an hour. Before and after cycling their spasticity was tested with an isokinetic dynamometer. The subjects were tested one week later by exactly the same procedure with a half an hour break instead of the cycling. Subjects were asked about their spasticity before and after the cycling and break. RESULTS: There was no significant difference in elicited peak torque either before and after the cycling, or before and after the break (MANOVA, P<0.05). Six out of 10 subjects estimated their spasticity as less after the cycling. CONCLUSION: With the isokinetic dynamometer, it was not possible to show an effect of passive cycling on spasticity reduction. However, six out of 10 of the subjects estimated their spasticity to be less after cycling. This positive effect might be attributed to a reduced spasticity in the trunk and/or to the attention the subjects perceived during the intervention. PMID- 15824756 TI - Recommended guidelines for studies of human subjects with spinal cord injury. PMID- 15824757 TI - A review of body mass index and waist circumference as markers of obesity and coronary heart disease risk in persons with chronic spinal cord injury. AB - STUDY DESIGN: Literature review. BACKGROUND: Increased fat mass and coronary heart disease (CHD) are secondary complications of chronic spinal cord injury (SCI). In able-bodied populations, body mass index (BMI, body weight (kg)/height (m(2))) is a widely used surrogate marker of obesity and predictor of CHD risk. Waist circumference, an accurate and reproducible surrogate measure of abdominal visceral adipose tissue, is also associated with CHD risk (more so than BMI) in able-bodied populations. OBJECTIVE: To review the literature on the accuracy of BMI and waist circumference as surrogate measures of obesity and CHD risk in persons with chronic SCI. SETTING: Ontario, Canada. METHODS: Literature review. RESULTS: In the SCI population, BMI is an insensitive marker of obesity, explains less of the variance in measured percent fat mass than in the able-bodied, and is inconsistently related to CHD risk factors. This may be due to potential measurement error, and to the inability of BMI to distinguish between fat and fat free mass and to measure body fat distribution. Waist circumference has not been validated as a surrogate measure of visceral adipose tissue, however preliminary evidence supports a relationship between waist circumference and CHD risk in the SCI population. CONCLUSIONS: We recommend that SCI-specific BMI classifications be determined. We also recommend that accuracy and reliability of waist circumference as a surrogate measure of visceral adipose tissue and CHD risk be determined in men and women with long-standing paraplegia and tetraplegia. PMID- 15824758 TI - Rehabilitation outcomes following traumatic spinal cord injury in a tertiary spinal cord injury centre: a comparison with an international standard. AB - STUDY DESIGN: Retrospective descriptive analysis of data of patients with traumatic spinal cord injury (SCI) in a tertiary SCI centre. OBJECTIVES: To identify the characteristics of the rehabilitation outcomes of patients with different levels of traumatic SCI and to compare the results with data reported in the American Consortium for Spinal Cord Medicine. SETTING: A newly established tertiary SCI centre in Tai Po Hospital, Tai Po, Hong Kong. METHODS: A total of 33 patients with traumatic SCI admitted in 2002 were included in the study. They were classified into different ASIA subgroups based on their levels and completeness of injury. The functional status changes measured by the Functional Independence Measure (FIM) (on admission, placement and upon discharge, and at 1 and 3 months post discharge) and discharge placement were recorded as rehabilitation outcomes. RESULTS: A total, 24 patients were tetraplegic while nine were paraplegic. Seven and two from tetraplegic and paraplegic groups were readmitted with late complications due to urinary tract infection, spasticity and/or occurrence of pressure sores. The mean age was found to be 48.36 (SD=15.64) years. In all, 16 (48.48%) sustained the injury from falling from height. The trend of FIM motor scores at discharge across different ASIA subgroups appeared to be comparable to those reported in the American Consortium for Spinal Cord Medicine with scores generally lower. Significant functional improvements during the hospital phase were found in the two tetraplegic and paraplegic ASIA D subgroups (t3=3.430, P<0.05; t2=4.083, P=0.55, respectively). Significant differences were also revealed among subgroups (F(7,32)=6.625, P<0.0005) with lower level tetraplegic groups appearing to stay much longer in the rehabilitation centre. In all, 64.5% of newly diagnosed patients returned to live in the community. CONCLUSIONS: This report gives a preliminary overview on the characteristics of rehabilitation outcomes in one of the SCI centres in Hong Kong in relation to the international standard. Longitudinal study with larger population and community integration outcomes might be included in the future to reveal a better picture in the SCI rehabilitation in Hong Kong. PMID- 15824760 TI - Antioxidant effects of statins. AB - Statins, a group of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, are widely used in clinical practice for their efficacy in producing significant reductions in plasma cholesterol and LDL cholesterol and in reducing morbidity and mortality from cardiovascular disease. However, several large clinical trials have suggested that the cholesterol-lowering effects of statins may not completely account for the reduced incidence of cardiovascular disease seen in patients receiving statin therapy. A number of recent reports have shown that statins may also have important antiinflammatory effects, in addition to their effects on plasma lipids. Since inflammation is closely linked to the production of reactive oxygen species (ROS), the molecular basis of the observed antiinflammatory effects of statins may relate to their ability block the production and/or activity of ROS. In this review, we will discuss both the inhibition of ROS generation by statins, through interference with NAD(P)H oxidase expression and activity, and the actions of statins that serve to blunt the damaging effects of these radicals, including effects on antioxidant enzymes, lipid peroxidation, LDL cholesterol oxidation and nitric oxide synthase. These antioxidant effects of statins likely contribute to their clinical efficacy in treating cardiovascular disease as well as other chronic conditions associated with increased oxidative stress in humans. PMID- 15824759 TI - Patterns of lower extremity innervation in pediatric spinal cord injury. AB - STUDY DESIGN: Retrospective review. OBJECTIVES: To identify relationships between lower extremity innervation and level of injury, mechanism of injury, and age at injury in a pediatric population with spinal cord injury (SCI). Secondarily, relationships between innervation and completeness of injury, time since injury, race, and sex were evaluated. SETTING: Pediatric orthopedic referral hospital, Philadelphia, Pennsylvania. METHODS: Records of 190 subjects, ages 1-21 years, were reviewed. Data collected from the medical record included lower extremity muscle innervation, American Spinal Injury Association (ASIA) level and class, mechanism of injury, age at injury, time since injury, race, and sex. To determine innervation, lower extremity muscles had been tested using surface electrical stimulation and identified as being innervated or denervated. If a muscle responded weakly, strength duration testing was performed. For analysis via logistic regression, subjects were grouped based upon level and mechanism of injury. RESULTS: A relationship (P<0.0001) was found between ASIA level and lower extremity innervation of all muscles and between length of time since injury and lower extremity innervation for some muscles. Following multiple logistic regression, only ASIA level remained as an independent predictor of lower extremity innervation status. CONCLUSION: Our results show that lower extremity innervation does differ based on the level of the injury. Denervation began to be seen with injuries in the lower thoracic region and more predominantly with injuries in the lumbar region. This supports our hypothesis that the incidence of lower motor neuron injuries would increase as injuries became more caudal. Our hypotheses of a relationship between innervation status and mechanism of injury and age at injury were not supported. This information is important in determining treatment strategies, eligibility for electrical stimulation techniques, and potential regenerative strategies. SPONSORSHIP: This study was funded by Shriners Hospitals for Children, Grant #8530. PMID- 15824761 TI - Antihypertensive drug interactions. AB - There are many drug interactions with antihypertensive agents and some of these are highly significant. Patients with hypertension frequently take multiple medications and may be at increased risk for drug interactions. Nearly every elderly patient with multiple medical problems will have the potential for one drug interaction in their regimen. These drug interactions can lead to morbidity or even mortality if appropriate steps are not taken to minimize this risk. Drug interactions may occur due to pharmacokinetic (i.e., absorption, distribution, metabolism, elimination) or pharmacodynamic interactions. Physicians and pharmacists must remain vigilant in their monitoring of potential drug interactions and make appropriate dosage or therapy adjustments. PMID- 15824762 TI - Modulation of the inflammatory process by statins. AB - Statins reduce cholesterol levels through competitive inhibition of 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase, the key enzyme that regulates cholesterol synthesis. The cholesterol-lowering effect of statins is also due to an increase in the uptake of cholesterol by cells as a result of intracellular cholesterol depletion and enhanced expression of low-density lipoprotein (LDL) receptors. The use of statins as lipid-lowering agents has lead to remarkable changes in the treatment and prevention of ischemic heart disease. Results of large clinical trials of patients with ischemic heart disease have demonstrated that statins reduce inflammatory markers such as C-reactive protein, an independent risk factor in the disease. Statins exhibit properties that are beyond their lipid-lowering effects. These non-lipid-lowering properties involve the inhibition of the isoprenoid pathway through decreased synthesis of many nonsteroidal isoprenoid compounds. The focus on the immunomodulatory effect of statins is the result of the positive outcome of pravastatin treatment in cardiac transplantation patients, as well as angiographic regression studies showing insignificant changes in the degree of coronary stenosis despite a large reduction in cardiac events. Statin treatment reduces the risk of ischemic stroke despite the fact that LDL cholesterol is not directly associated with the risk of stroke. This observation lead to the investigation of the role of statins in inflammation and the immune system. Recent research data demonstrated that statins inhibit the induction of the major histocompatibility (MHC) class II expression by interferon-gamma (IFN-gamma), leading to repression of MHC II mediated T-cell activation. Furthermore, statins inhibit the expression of specific cell surface receptors on monocytes, adhesion molecules and also integrin-dependent leucocyte adhesion. While statins may stimulate the secretion of caspase-1, IL-1beta and IL-18 in peripheral mononuclear cells in response to Mycobacterium tuberculosis, they exhibit additional effects on inflammation by decreasing IL-6 synthesis in human vascular smooth muscle cells (VSMC) in vitro. The focus of this monograph is to highlight the role of statins in the modulation of the immune system and inflammatory processes. PMID- 15824768 TI - "GaI": a versatile reagent for the synthetic chemist. AB - The current renaissance in main group chemistry has been fuelled by the remarkable array of fundamentally interesting yet synthetically applicable low oxidation state p-block compounds that have appeared over the last decade. Their syntheses generally require the ready availability of low oxidation state element halide precursors. In the case of gallium this is provided by the simple to prepare reagent, "GaI", which since it was first reported in 1990, has been utilised in areas as varied as organic synthesis and gallium cluster construction. This article tracks the history of this extraordinary material and highlights its synthetic diversity; hopefully allowing the reader to envisage its application to aspects of their own research fields. PMID- 15824769 TI - Interaction of cobalt(II) and nickel(II) with a gamma-linked, bis-beta-diketone ligand to yield an unusual coordination motif. AB - Discrete cobalt(II) and nickel(II) complexes of type ML (where L is a bis(2,4 pentanedionate) derivative incorporating a -SCH2CH2S- bridge between the gamma carbons of respective beta-diketone units) have been synthesised and represent the first examples in which a gamma-substituted, thioether-containing beta diketone ligand coordinates via a sulfur and oxygen donor in preference to the classical coordination mode involving both beta-diketonate oxygens. PMID- 15824770 TI - Magnetic ordering in a mononuclear cobalt(II) complex containing a Schiff-base pentadentate ligand. AB - A mononuclear cobalt(II) complex CoL5, containing the pentadentate O2N3 salen type Schiff-base ligand H2L5=N,N[prime or minute]-bis(3-tert-butyl-2-hydroxy-5 methylbenzylidenyl)-1,7-diamino-4-methyl-4-azaheptane, exhibits magnetic ordering at 4 K as proven by ac magnetic susceptibility (both in- and out-of-phase), magnetization, field-cooled magnetization and zero-field cooled magnetization measurements. PMID- 15824771 TI - The first examples of metal-mediated addition of a phosphorus imine to nitriles; the preparation and X-ray crystal structures of [PtCl4{NH=C(Et)N=PPh3}2] and [PtCl2(EtCN){NH=C(Et)N[double bond, length as m-dash]PPh3}]. AB - Imino(triphenyl)phosphorane, Ph3P=NH (1), reacts with nitrile complexes of Pt(IV) to generate hydrolytically sensitive [PtCl4{NH=C(R)N=PPh3}2](R=Me 2a, Et 2b, Ph 2c), and with the Pt(II) complex [PtCl2(EtCN)2] to give [PtCl2(EtCN){NH=C(Et)N=PPh3}](3) and [PtCl2{NH=C(Et)N=PPh3}2](4); X-ray crystallography performed upon (2b) and (3) confirms the presence of an imine/nitrile addition ligand bound by the terminal nitrogen. PMID- 15824772 TI - Synthesis, crystal structures and magnetic properties of cyanide- and phenolate bridged [M(III)NiII]2 tetranuclear complexes (M=Fe and Cr). AB - The binuclear complex NiII2L(H2O)2(ClO4)2(1) and the neutral tetranuclear bimetallic compounds [{M(III)(phen)(CN)4}2{NiII2L(H2O)2}].2CH3CN with M=Fe (2) and Cr (3)[H2L=11,23-dimethyl-3,7,15,19-tetraazatricyclo[19.3.1.1(9,13)]hexacosa 2,7,9,11,13(26),14,19,21(25),22,24-decaene-25,26-diol] have been synthesized and the structures of and determined by single crystal X-ray diffraction. and are isostructural compounds whose structure is made up of centrosymmetric binuclear cations [Ni2(L)(H2O)2]2+ and two peripheral [M(phen)(CN)4]- anions [M=Fe (2) and Cr (3)] acting as monodentate ligands towards the nickel atoms through one of their four cyanide nitrogen atoms. The environment of the metal atoms in 2 and 3 is six-coordinated: two phen-nitrogen and four cyanide-carbon atoms at the iron and chromium atoms and a water molecule, one cyanide-nitrogen and two phenolate oxygens and two imine-nitrogens from the binucleating ligand L2- at the nickel atom build distorted octahedral surroundings. The values of the FeNi and CrNi separations through the single cyanide bridge are 5.058(1) and 5.174(2)A respectively, whereas the Ni-Ni distances across the double phenolate bridge are 3.098(2)(2) and 3.101(1) A (3). The magnetic properties of have been investigated in the temperature range 1.9-290 K. The magnetic behaviour of corresponds to that of an antiferromagnetically coupled nickel(II) dimer with J=-61.0(1) cm-1, the Hamiltonian being defined as H=-J S(A).S(B). An overall antiferromagnetic behaviour is observed for and with a low-lying singlet spin state. The values of the intramolecular magnetic couplings are J(Fe-Ni)=+17.4(1) cm-1 and J(Ni-Ni(a))= 44.4(1) cm-1 for and J(Cr-Ni)=+11.8(1) cm-1 and J(Ni-Ni(a))=-44.6(1) cm-1 for [H= J(M-Ni)(S(M).S(Ni)+S(Ma).S(Nia))-J(Ni-Nia)S(Ni)S(Nia)]. Theoretical calculations using methods based on density functional theory (DFT) have been employed on in order to analyze the efficiency of the exchange pathways involved and also to substantiate the exchange coupling parameters. PMID- 15824773 TI - A series of neutral radical CpNi(dithiolene). complexes. AB - The heteroleptic neutral radical dithiolene complexes CpNi(dmit)., CpNi(dsit). and CpNi(dmid).(dmit=1,3-dithiole-2-thione-4,5-dithiolate; dsit=1,3-dithiole-2 thione-4,5-diselenolate; dmid=1,3-dithiole-2-one-4,5-dithiolate) are obtained from the reaction of (Cp2Ni)BF4 with either (n-Bu4N)[Ni(dmit)2] and (n Bu4N)[Ni(dmid)2] or PhSb(dmit) and PhSb(dsit), respectively. The three complexes reduce reversibly to the corresponding Ni(II) anions and oxidize reversibly to the cationic state. As deduced from DFT calculations performed on CpNi(dmit)., the SOMO of these complexes is essentially localized on the dithiolene moiety with little metal contribution. CpNi(dsit). is isostructural with CpNi(dmit). and crystallizes in the monoclinic system, space group P2(1). In the solid-state structures of both CpNi(dmit). and CpNi(dsit)., molecules interact through a three-dimensional set of intermolecular interactions mediated by short SS, SeSe and SSe contacts, as confirmed from the temperature and field dependence of the magnetic susceptibility by the observation of an antiferromagnetic ground state below T(Neel)=27 K in CpNi(dmit)., 18 K in CpNi(dsit).. Finally, CpNi(dmid). crystallizes in the orthorhombic system, space group Pnma. Molecules organize into uniform chains through the stacking of the dmid moieties in a sigma-type face-to-face overlap. PMID- 15824774 TI - Reactions of a {Mo16}-type polyoxometalate cluster with electrophiles: a synthetic, theoretical and magnetic investigation. AB - A medium-nuclearity mixed-valence polyoxomolybdate [H2Mo16O52]10-={Mo16}(1a) was synthesized using an approach that employed protonated hexamethylenetetramine (HMTAH+) as counter ion and yielded (HMTAH)10 1a.34 H2O (1). The {Mo16} cluster anion exhibits significant nucleophilicity and traps electrophiles such as divalent transition metal ions, resulting in a family of isostructural compounds based on {Mo16M2}-type anions [M(H2O)8H2Mo16O52]6- (M=FeII (2), MnII (3), CoII (4)). The highly reactive nature of the {Mo16} system is also revealed by rearrangement and decomposition reactions of to either slowly form a sodium bridged heptamolybdate-based chain compound (5) when left in the reaction solution or, in the presence of very high concentrations of electrophiles, to heptamolybdate-based cluster compounds [M2(H2O)9Mo7O24]2- of the {M2Mo7}-type (M=FeII (6), MnII (7)). Compounds were characterised by single crystal X-ray diffraction, elemental analysis, IR spectroscopy, magnetic susceptibility measurements, and density functional theory calculations. PMID- 15824775 TI - Crystal structure and magnetic properties of a pseudo-cubic close-packed array of oxalate linked {FeII6(mu3-OH)6}6+ clusters. AB - The ionic hexanuclear cluster aggregate [FeII6(mu3-OH)6]6+ has been synthesised using hydrothermal conditions starting from ferrous oxalate in the presence of barium and bromide or iodide ions. A single crystal X-ray structure on the compound Ba4[FeII6(mu3-OH)6(C2O4)6]Br2.6H2O shows that the [FeII6(mu3-OH)6]6+ units are held together by bridging oxalates in a pseudo-cubic close-packed array. The barium ions in conjunction with oxalate groups provide a barrel-shaped cage between the FeII6 aggregates containing the bromide counterions and lattice waters and corresponding to an 'octahedral hole' in the pseudo-cubic close-packed structure. A magnetic susceptibility study shows that the FeII centres are antiferromagnetically coupled. Below 10 K the system displays a long range antiferromagnetic ordering mediated by the oxalate bridges and a molecular-based description of the magnetism is no longer valid. PMID- 15824776 TI - Synthesis and characterization of mono beta-diketiminatosamarium amides and hydrocarbyls. AB - Reaction of SmCl3 with 1 eq of KL (L =[DippNC(Me)CHC(Me)NDipp]; Dipp = 2,6-i Pr2C6H3) in THF afforded the dimeric samarium dichloride LSmCl2(THF)Cl2SmL (1) in high yield. Reactions of 1 with NaN(SiMe3)2, KNHAr (Ar = 2,4,6-t-Bu3C6H2), KBHEt3, and KCp*(Cp*= C5Me5) yielded various new complexes: LSmClN(SiMe3)2 (2), LSm[N(SiMe3)2]2 (3), LSmNHAr(HBEt3) (4), LSm(NHAr)2 (5), and LSmCp*Cl (6). Reaction of 1 with one eq of NaN(SiMe3)2 followed by treatment with excess AlMe3 afforded a unique bimetallic samarium tetramer Cl3L2Sm2(AlMe4)2Sm2L2Cl3 (7). Reaction of 6 with LiMe or LiCH2SiMe3 afforded LSmCp*Me (8) and LSmCp*CH2SiMe3 (9) in excellent yield. Methyl abstraction from with B(C6F5)3 in toluene yielded the cationic borate species (LSmCp*)[MeB(C6F5)3] (10). Molecular structures of 1 7 and 9 were determined by X-ray single crystal analysis. PMID- 15824777 TI - "A posteriori" modification of carbosilane dendrimers and dendrons: their activation in core and branch positions. AB - The introduction of phenyl groups at different points on carbosilane dendrimers allows their acidolytic conversion to highly reactive triflato groups which in turn are readily substituted by anionic nucleophiles. Core phenylated first fourth generation dendrimers were synthesized from tri(allyl)phenylsilane by an alternating sequence of hydrosilylation and allylation steps. Similarly, carbosilane dendrimers containing phenyl-Si groups at the branching points and in the periphery were prepared from tetraallylsilane which was hydrosilylated with PhHSiCl2. Reaction of the phenylated dendrimers with triflic acid in toluene cleanly gave the silyl triflate derivatives, provided that the correct stoichiometry of the reagents was used. In the presence of a large excess of triflic acid the SiMe3-end groups are slowly converted to SiMe2(OTf)-units. The proof of concept was provided by the fixation of a {Ph2PCH2} group using the lithiated diphenylphosphinomethanide Ph2PCH2Li, obtained by cleavage of Ph3SnCH2PPh2 with PhLi, as well as a lithiated ether-alcohol functionalized triphos derivative to the core of a third generation carbosilane dendrimer. PMID- 15824778 TI - The fixation of bis(2-pyridylimino)isoindolato (BPI) ligands to dendritic carbosilanes. AB - Bis(2-pyridylimino)isoindolato (BPI) ligands, containing an alkynyl linker unit which allows their fixation to carbosilane dendrimers and dendrons, were synthesized by reaction of 4-nitrophthalodinitrile with 4-butynol giving the phthalodinitrile derivative containing the linker. These were subsequently reacted with two molar equivalents of 2-amino-4-methylpyridine and 2-amino-4 (t)butylpyridine yielding the respective BPI protioligands 2a and 2b. Lithiation with LDA and reaction with Si-Cl or Si-OTf (OTf=triflate) end groups in core or peripheral positions of dendritic carbosilanes gave the endodendrally and expdendrally functionalized dendrimers. Among these the first and second generation dendrimers [G-1](8-exo)-4-[C[triple bond, length as m-dash]CCH2CH2O] 10-MeBPI (8), [G-1]12-exo-4-[C[triple bond, length as m-dash]CCH2CH2O]-10-MeBPI (9) and [G-2](16-exo)-4-[C[triple bond, length as m-dash]CCH2CH2O]-10-MeBPI (10) were synthesized and fully characterized. The functional dendrimers were metallated by reaction with [(PhCN)2PdCl2] in dichloromethane to give the corresponding pallada-dendrimers. PMID- 15824779 TI - Electronic and steric effects in fragmentation reactions of Os3(CO)9(mu-C4Ph4). AB - P-Donor nucleophiles of cone angle >or=145 degrees are known to react with Os3(CO)9(mu-C4Ph4) quite differently from those with cone angles M3+ can be observed. In the binuclear complexes the coordinatively saturated octahedral ion M[prime or minute] is bound to the two oxygen atoms of the bridging oximate groups and the four nitrogen atoms of the tetradentate ligand tren. Two metal ions (M and M') are linked by the double cis-oximate bridge and are incorporated in a six-membered bimetallic chelate ring. Metallamacrocycle formation leads to certain changes in the structural parameters of the binuclear complexes as compared to those observed in the mononuclear species. Also the study of the electrochemical activity of binuclear complexes has shown important differences in their redox behaviour as compared to their mononuclear precursors. PMID- 15824782 TI - Mononuclear oxovanadium complexes of tris(2-pyridylmethyl)amine. AB - Mononuclear oxovanadium(IV) and dioxovanadium(V) complexes of tris(2 pyridylmethyl)amine (tpa) have been prepared for the first time. Crystal structure determinations of three oxovanadium(IV) complexes, [VO(SO4)(tpa)], [VOCl(tpa)]PF6, or [VOBr(tpa)]PF6, and a dioxovanadium(v) complex [V(O)2(tpa)]PF6 disclosed that the tertiary nitrogen of the tpa ligand always occupies the trans to-oxo site. The structures of an oxo-peroxo complex [VO(O2)(tpa)]Cl that was prepared previously and of a mu-oxo vanadium(III) complex [{VCl(tpa)}2(mu O)](PF6)2 have also been determined. The tertiary nitrogen is located at a trans site to the peroxo and chloride ligands, respectively. The total sums of the four V-N bond lengths from the tpa ligand are remarkably similar among the six complexes, indicating that the vanadium oxidation states become less influential in tpa bonding due primarily to the coordination of electron-donating oxo ligand(s). Absorption spectra of [VOCl(tpa)]+ in acetonitrile showed a significant change upon addition of p-toluenesulfonic acid and HClO4, but not on addition of benzoic acid. Protonation at the oxo ligand by the former two acids is suggested. Cyclic voltammetric studies in acetonitrile verified the proton coupled redox behavior of the V(III)/V(IV) process involving the oxo ligand for the first time. From the dependence of the added p-toluenesulfonic acid to the CV, redox potentials for the following species have been estimated: [V(IV)OCl(tpa)]+/[V(III)OCl(tpa)](E1/2=-1.59 V vs. Fc+/Fc), [V(IV)(OH)Cl(tpa)]2+/[V(III)(OH)Cl(tpa)]+(Epc=-1.34 V), [V(IV)(OH2)Cl(tpa)]3+/[V(III)(OH2)Cl(tpa)]2+(Epa=-0.49 V), and [V(IV)Cl2(tpa)]2+/[V(III)Cl2(tpa)]+(E1/2=-0.89 V). The reduction of [V(V)(O)2(tpa)]+ in 0.05 M [(n-Bu)4N]PF6 acetonitrile showed a major irreversible reduction wave V(V)/(IV) at -1.48 V. The metal reduction potentials of the oxovanadium(IV) and dioxovanadium(V) species are very close, reinforcing the significant influence of the oxo ligand(s). PMID- 15824783 TI - New and versatile routes to zirconium imido dichloride compounds. AB - Reactions of zirconium dialkyl- or bis(amido)-dichloride complexes "[Zr(CH2SiMe3)2Cl2(Et2O)2]" or [Zr(NMe2)2Cl2(THF)2] with primary alkyl and aryl amines are described. Reaction of "[Zr(CH2SiMe3)2Cl2(Et2O)2]" with RNH2 in THF afforded dimeric [Zr2(mu-NR)2Cl4(THF)4](R=2,6-C6H3iPr2 (1), 2,6-C6H3Me2 (2) or Ph (3)), [Zr2(mu-NR)2Cl4(THF)3](R=tBu (5), iPr (6), CH2Ph (7)), or the "ate" complex [Zr2(mu-NC6F5)2Cl6(THF)2{Li(THF)3}2](4, the LiCl coming from the in situ prepared "[Zr(CH2SiMe3)2Cl2(Et2O)2]"). With [Zr(NMe2)2Cl2(THF)2] the compounds [Zr2(mu NR)2Cl4(L)x(L')y](R=2,6-C6H3iPr2 (8), 2,6-C6H3Me2 (9), Ph (10) or C6F5 (11); (L)x(L')y=(NHMe2)3(THF), (NHMe2)2(THF)2 or undefined), [Zr2(mu-NtBu)2Cl4(NHMe2)3] (12) and insoluble [Zr(NR)Cl2(NHMe2)]x(R=iPr (13) or CH2Ph (14)) were obtained. Attempts to form monomeric terminal imido compounds by reaction of or with an excess of pyridine led, respectively, to the corresponding dimeric adducts [Zr2(mu-2,6-C6H3Me2)2Cl4(py)4] (15) and [Zr2(mu-NtBu)2Cl4(py)3] (16). The X-ray structures of 1, 2, 4, 8, 12 and 15 have been determined. PMID- 15824784 TI - Pressure and temperature effects on metal-to-metal charge transfer in cyano bridged CoIII-FeII complexes. AB - The effects of pressure and temperature on the energy (Eop) of the metal-to-metal charge transfer (MMCT, FeII-->CoIII) transition of the cyano-bridged complexes trans-[L14CoNCFe(CN)5]- and cis-[L14CoNCFe(CN)5]- (where L14=6-methyl-1,4,8,11 tetraazacyclotetradecan-6-amine) were examined. The changes in the redox potentials of the cobalt and iron metal centres with pressure and temperature were also examined and the results interpreted with Marcus-Hush theory. The observed redox reaction volumes can mainly be accounted for in terms of localised electrostriction effects. The shifts in Eop due to both pressure and temperature were found to be less than the shifts in the energy difference (DeltaE degrees]) between the CoIII-FeII and CoII-FeIII redox isomers. The pressure and temperature dependence of the reorganisational energy, as well as contributions arising from the different spin states of CoII, are discussed in order to account for this trend. To study the effect of pressure on CoIII electronic absorption bands, a new cyano-bridged complex, trans-[L14CoNCCo(CN)5], was prepared and characterised spectroscopically and structurally. X-Ray crystallography revealed this complex to be isostructural with trans-[L14CoNCFe(CN)5].5H2O. PMID- 15824785 TI - C-C coupling reactions in the coordination sphere of rhodium(I) and rhodium(III): New routes for the di- and trimerization of terminal alkynes. AB - The alkynyl(vinylidene)rhodium(I) complexes trans-[Rh(C[triple bond, length as m dash]CR)(=C=CHR)(PiPr3)2] 2, 5, 6 react with CO by migratory insertion to give stereoselectively the butenynyl compounds trans-[Rh{eta1-(Z)-C(=CHR)C[triple bond, length as m-dash]CR}(CO)(PiPr3)2](Z)-7-9, of which (Z)-7 (R=Ph) and (Z)-8 (R=tBu) rearrange upon heating or UV irradiation to the (E) isomers. Similarly, trans-[Rh{eta1-C(=CH2)C[triple bond, length as m-dash]CPh}(CO)(PiPr3)2] 12 and trans-[Rh{eta1-(Z)-C(=CHCO2Me)C[triple bond, length as m-dash]CR}(CO)(PiPr3)2](Z) 15, (Z)-16 have been prepared. At room temperature, the corresponding "non substituted" derivative trans-[Rh{eta1-C(=CH2)C[triple bond, length as m dash]CH}(CO)(PiPr3)2] 18 is in equilibrium with the butatrienyl isomer trans [Rh(eta1-CH=]C=C=CH2)(CO)(PiPr3)2] 19 that rearranges photochemically to the alkynyl complex trans-[Rh(C[triple bond, length as m-dash]CCH=CH2)(CO)(PiPr3)2] 20. Reactions of (Z)-7, (E)-7, (Z)-8 and (E)-8 with carboxylic acids R'CO2H (R'=CH3, CF3) yield either the butenyne (Z)- and/or (E)-RC[triple bond, length as m-dash]CCH=CHR or a mixture of the butenyne and the isomeric butatriene, the ratio of which depends on both R and R'. Treatment of 2 (R=Ph) with HCl at -40 degrees C affords five-coordinate [RhCl(C[triple bond, length as m-dash]CPh){(Z) CH=CHPh}(PiPr3)2] 23, which at room temperature reacts by C-C coupling to give trans-[RhCl{eta2-(Z)-PhC[triple bond, length as m-dash]CCH=CHPh}(PiPr3)2](Z)-21. The related compound trans-[RhCl(eta2-HC[triple bond, length as m dash]CCH=CH2)(PiPr3)2] 27, prepared from trans-[Rh(C[triple bond, length as m dash]CH)(=C=CH2)(PiPr3)2] 17 and HCl, rearranges to the vinylvinylidene isomer trans-[RhCl(=C=CHCH=CH2)(PiPr3)2] 28. While stepwise reaction of 2with CF3CO2H yields, via alkynyl(vinyl)rhodium(III) intermediates (Z)-29 and (E)-29, the alkyne complexes trans-[Rh(kappa1-O2CCF3)(eta2-PhC[triple bond, length as m dash]CCH=CHPh)(PiPr3)2](Z)-30 and (E)-30, from 2 and CH3CO2H the acetato derivative [Rh(kappa2-O2CCH3)(PiPr3)2] 33 and (Z)-PhC[triple bond, length as m dash]CCH=]CHPh are obtained. From 6 (R=CO2Me) and HCl or HC[triple bond, length as m-dash]CCO2Me the chelate complexes [RhX(C[triple bond, length as m dash]CCO2Me){kappa2(C,O)-CH=CHC(OMe)=O}(PiPr3)2] 34 (X=Cl) and 35 (X=C[triple bond, length as m-dash]CCO2Me) have been prepared. In contrast to the reactions of [Rh(kappa2-O2CCH3)(C[triple bond, length as m-dash]CE)(CH=CHE)(PiPr3)2] 37(E=CO2Me) with chloride sources which give, via intramolecular C-C coupling, four-coordinate trans-[RhCl{eta2-(E)-EC[triple bond, length as m dash]CCH=CHE}(PiPr3)2](E)-36, treatment of 37with HC[triple bond, length as m dash]CE affords, via insertion of the alkyne into the rhodium-vinyl bond, six coordinate [Rh(kappa2-O2CCH3)(C[triple bond, length as m-dash]CE){eta1-(E,E) C(=CHE)CH=CHE}(PiPr3)2] 38. The latter reacts with MgCl2 to yield trans [RhCl{eta2-(E,E)-EC[triple bond, length as m-dash]CC(=CHE)CH=CHE}(PiPr3)2] 39, which, in the presence of CO, generates the substituted hexadienyne (E,E) EC[triple bond, length as m-dash]CC(=CHE)CH=CHE 40. PMID- 15824786 TI - Metal-to-ligand charge-transfer sensitisation of near-infrared emitting lanthanides in trimetallic arrays M2Ln (M=Ru, Re or Os; Ln=Nd, Er or Yb). AB - The new pro-ligand 4-methyl-4'-(carbonylamino(2-(tert-butoxycarbonylamino)ethyl)) 2,2'-bipyridyl (L1) has been prepared and used to synthesise the complex fac Re(I)Cl(CO)3(L1) 1 and the complex salts [M(II)(bipy)2(L1)](PF6)2 (M=RuII 8 or OsII 15). Deprotection with trifluoroacetic acid affords the amine-functionalised derivatives fac-Re(I)Cl(CO)3(L2) 2, [M(II)(bipy)2(L2)](PF6)2 (M=RuII 9 or OsII 16) which react with the dianhydride of diethylenetriamine pentaacetic acid (DTPA) to give the binuclear complex {fac-Re(I)Cl(CO)3}2(L3) 3 and the complex salts [{M(II)(bipy)2}2(L3)](PF6)4 (M = RuII 10 or OsII 17). The latter react with salts Ln(OTf)3 to afford a series of 12 heterotrimetallic compounds that contain a lanthanide (Ln) ion in the DTPA binding site; {fac-Re(I)Cl(CO)3}2(L3)LnIII (Ln=Nd 4, Er 5, Yb 6 or Y 7) and [{M(II)(bipy)2}2(L3)LnIII](PF6)(OTf)3 (M=RuII, Ln=Nd 11, Er 12, Yb 13 or Y 14; M=OsII, Ln=Nd 18, Er 19, Yb 20 or Y 21). All of these trimetallic species display absorption bands ascribed to metal-to-ligand charge-transfer (MLCT) excitations, and luminescence measurements show that these excited states can be used to sensitise near-infrared emission from LnIII (Ln=Nd, Er or Yb) ions. Single crystal X-ray structures of L1 and [RuII(bipy)2(L2H)](H2PO4)3.(CH3)2CO.0.8H2O were obtained, the latter revealing the presence of H2PO4- counter anions, the source of which is presumed to be hydrolysis of PF6- ions. PMID- 15824787 TI - Bulky triarylarsines are effective ligands for palladium catalysed Heck olefination. AB - The four arsines, As{C6H3(o-CH3)(p-Z)}3{Z=H (2a) or OMe (2b)} and As{C6H3(o CHMe2)(p-Z)}3{Z=H (2c) or OMe (2d)} react with [PdCl2(NCPh)2] or [PtCl2(NCBu(t))2] to give trans-[MCl2L2] or trans-[M2Cl2(mu-Cl)2L2]. The crystal structures of trans-[PdCl2(2a)2] and [PtCl2(2d)2] have been determined, the latter as its dichloromethane solvate. The structures show that in these complexes, the ligands adopt gga type conformations as do all analogous tri-o tolyl- and tri-o-isopropylphenylphosphines in square-planar and octahedral complexes. The variable-temperature NMR behaviour of the complexes shows that they are fluxional due to restricted As-C bond rotation. The rate of the fluxionality is more rapid than in the analogous phosphine complexes and this is associated with longer As-C and As-M bonds allowing more free movement. The catalytic activity of the palladium complexes of the arsines and their phosphine analogues for the reaction of 4-bromoacetophenone and n-butyl acrylate has been screened. The results show that the arsines are generally superior to the phosphines as ligands for this catalysis. Tri(o-isopropylphenyl)phosphine and tri(o-isopropylphenyl)arsine are superior to tri-o-tolylphosphine as ligands for this Heck reaction and a p-methoxy substituent improves the arsine catalyst but not the phosphine catalyst. The phosphine catalysts are superior to the arsine catalysts for the reaction of 4-chloroacetophenone and n-butyl acrylate. These observations are discussed in the context of ligand stereoelectronic effects, as measured by the Tolman electronic parameter, nuCO of the [NiL(CO)3]{L=AsAr3 or PAr3}. PMID- 15824788 TI - Macropolyhedral boron-containing cluster chemistry. The reaction of B16H20 and B14H18 with [PtMe2(PMe2Ph)2] to give [(PMe2Ph)2PtB16H17Me] and [(PMe2Ph)2PtB14H16]. AB - Structurally characterised 17-vertex [(PMe2Ph)2PtB16H17Me] 3 is obtained, albeit in low yield, by platination of 16-vertex B16H20 1 using [PtMe2(PMe2Ph)2] under mild conditions. Platination has occurred on the {B10} subcluster of 1, interesting in that B16H20 itself deprotonates on the {B8} subcluster: the reference 16-vertex [B16H19]- anion 1a, prepared by deprotonation of 1 with 1,8 bis(dimethylamino)naphthalene, is also structurally characterised. [PtMe2(PMe2Ph)2] with 14-vertex B14H18 2 similarly gives a low yield of 15-vertex [(PMe2Ph)2PtB14H16] 5, of formulation and structure substantiated by DFT calculations. PMID- 15824789 TI - Porphyrin-triarylamine conjugates: strong electronic communication between triarylamine redox centers via the porphyrin dication. AB - A set of porphyrin-triarylamine hybrids have been synthesized in good yield by Sonogashira palladium-catalyzed cross-coupling reactions between the zinc complex of 5,15-diethynyl-10,20-dimesitylporphyrin and the appropriate iodophenyldiarylamines. The crystal structure of porphyrin 1 shows that the dihedral angle between the acetylene-bonded benzene rings and the porphyrin macrocycle is 20.0 degrees. Such a structural characteristic enables effective electronic perturbations within the molecule. The electronic spectra are red shifted and display a broad Soret band and an intense Q band relative to those of meso-substituted tetraarylporphyrins. These conjugates display four oxidations and one reduction. All the electrochemical reactions involve one-electron transfer. The first and second oxidations are reversible and can be assigned to the porphyrin-centered reactions. The third and fourth ones, separated by about 270 mV, correspond to the triarylamine units. The comproportionation constant (Kc) is calculated to be 3.67x10(4). The electron coupling between the triarylamine moieties, at a separation of >23 A, is remarkably strong. The electrochemical results and the absorption spectra show that the electronic characteristics of these porphyrins can be significantly modulated by the triarylamine substituents via the conjugated carbon-carbon triple bond. Variations of the substituents on the triarylamines can fine-tune the electronic properties of these molecules. PMID- 15824790 TI - Syntheses, crystal structures and properties of novel copper(II) complexes obtained by reactions of copper(II) sulfate pentahydrate with tripodal ligands. AB - Three novel metal-organic frameworks (MOFs), [Cu(1)SO4].H2O (4), [Cu2(2)2(SO4)2].4H2O (5) and [Cu(3)(H2O)]SO4.5.5H2O (6), were obtained by hydrothermal reactions of CuSO4.5H2O with the corresponding ligands, which have different flexibility. The structures of the synthesized complexes were determined by single-crystal X-ray diffraction analyses. Complex 4 has a 2D network structure with two types of metallacycles. Complex 5 also has a 2D network structure in which each independent 2D sheet contains two sub-layers bridged by oxygen atoms of the sulfate anions. Complex 6 has a 2D puckered structure in which the sulfate anions serve as counter anions, which are different from those in complexes 4 (terminators) and 5 (bridges). The different structures of complexes 4, 5 and 6 indicate that the nature of organic ligands affected the structures of the assemblies greatly. The magnetic behavior of complex 5 and anion-exchange properties of complex 6 were investigated. PMID- 15824791 TI - Syntheses, structures and fluorescent properties of two novel coordination polymers in the U-Cu-H3pdc system. AB - Two novel coordination polymers, UO2(C5H2N2O4)(H2O) (1) and (UO2)Cu(C5H2N2O4)2(H2O)2 (2), have been prepared by the hydrothermal reaction of uranyl nitrate hexahydrate [(UO2(NO3)2.6H2O], 3,5-pyrazoledicarboxylic acid (H3pdc) and copper(II) nitrate hemipentahydrate (Cu(NO3)2.2.5H2O) and characterized by single-crystal X-ray diffraction, thermogravimetric analyses (TGA) and fluorescence spectroscopy. Compound 1 (monoclinic, P2(1)/c, a=6.9556(6)A, b=11.302(1)A, c= 10.5288(9)A, beta=90.057(2) degrees and Z=4) consists of a two-dimensional sheet containing uranyl hexagonal bipyramids. Compound 2 (triclinic, P-1, a=5.1014(7)A, b=7.6067(11)A, c=10.2910(15)A, alpha=72.380(3) degrees, beta=86.796(3) degrees, gamma=84.447(3) degrees and Z=1) consists of two-dimensional sheets. Both structures contain the linear UO2(2+) moiety and have extended networks built up from the H3pdc ligand. Compound 1 exhibits the characteristic UO(2)2+ emission spectra when it is excited at the ligand or uranium excitation wavelength. With the addition of the copper metal center in compound 2, the uranium emission is absent regardless of the excitation wavelength. PMID- 15824792 TI - Kinetics and mechanisms of the electron transfer reactions of oxo-centred carboxylate bridged complexes, [Fe3(mu3-O)(O2CR)6L3]ClO4, with verdazyl radicals in acetonitrile solution. AB - A range of oxo-centred, carboxylate bridged tri-iron complexes of general formula [Fe3(mu3-O)(O2CR)6L3]ClO4(R=CH2CN, CH2F, CH2Cl, CH2Br, p-NO2C6H4; L=pyridine, 3 methylpyridine, 4-methylpyridine, 3,5-dimethylpyridine, 3-cyanopyridine and 3 fluoropyridine) have been prepared and characterised. The choice of R and L was dictated by the requirement that the complexes undergo a one-electron reduction when reacted with verdazyl radicals. All except the complexes where L=pyridine and R=CH2CN, CH2Cl and p-NO2C6H4 have not been previously reported. The redox behaviour of these compounds has been investigated using cyclic voltammetry in acetonitrile in the absence and in the presence of free L. In general, all complexes exhibited reversible one-electron reductions. Electrochemical behaviour improved in the presence of an excess of L. The kinetics of the electron transfer reaction observed when acetonitrile solutions of the complexes were reacted with a range of verdazyl radicals were monitored using stopped-flow spectrophotometry. Under the experimental conditions, the reactions were quite rapid and were monitored under second-order conditions. Marcus linear free energy plots indicated that the outer-sphere electron transfer reactions were non-adiabatic in nature. Nevertheless, application of the self-exchange rate constants of the verdazyl radicals, k11, and the tri-iron complexes, k22, to the Marcus cross relation resulted in calculated values of the cross-reaction rate constant, k12, that were within a factor of five of the experimentally determined value. PMID- 15824793 TI - Synthesis and characterisation of a series of alkylmagnesium amide and related oxygen-contaminated "alkoxy" compounds. AB - Synthesised either by an unusual tert-butyl metathesis between tert-butyllithium and a n,s-butylmagnesium amide or by reaction of an alkyl Grignard reagent and a sodium amide, five tert-butylmagnesium amides, Bu(t)MgDBA (5)(DBA=dibenzylamide), Bu(t)MgDA (6)(DA=diisopropylamide), Bu(t)MgHMDS (7)(HMDS=1,1,1,3,3,3 hexamethyldisilazide), Bu(t)MgTMP (8)(TMP=2,2,6,6-tetramethylpiperidide) and Bu(t)MgNCy2 (9)(cy=cyclohexyl) have been isolated as crystalline solids. All five amides have been characterised by X-ray crystallography and solution NMR spectroscopic studies. The former studies reveal a common dimeric molecular structure with amido bridges in a planar (MgN)2 ring and terminal Bu(t) ligands on the Mg atoms. Also described is the dodecameric primary amide [Bu(n)MgN(H)Dipp]12 (10a) and its monomeric solvate Bu(n)MgN(H)Dipp.TMEDA (10b)(Dipp=2,6-diisopropylphenyl; TMEDA=N,N,N',N'-tetramethylethylenediamine). The crystal structures of the oxo-insertion products Bu(t)MgOBu(t).THF (11), Bu(t)Mg(mu-OBu(t))(mu-TMP)MgTMP (12) and Mg(OBu(n))HMDS.solv [solv=THF (13a) or Et2O (13b)], made fortuitously during the course of this work, are also presented. PMID- 15824794 TI - Calcium sulfate hemihydrate is the inorganic mineral in statoliths of Scyphozoan medusae (Cnidaria). AB - Scyphomedusae use inorganic crystals (statoliths) for gravity sensing. The organs which contain the statoliths are called rhopalia. Rhopalia of five different species of the three different orders of the class Scyphozoa were studied with high-end solid-state chemical methods to elucidate the crystallographic nature of the biomineral: synchrotron powder diffraction, synchrotron single-crystal diffraction, synchrotron microtomography, scanning electron microscopy, and energy dispersive X-ray spectroscopy. Each rhopalium contains a large number of statoliths in an ordered way. The statoliths of all species consist of calcium sulfate hemihydrate, a water-deficient phase. This is remarkable for sea-living organisms consisting mostly of water. The phylogenetic relationships within the class Scyphozoa are discussed. PMID- 15824797 TI - [Cataract in childhood: socioeconomic and gestational profiles and neuropsychomotor development]. AB - PURPOSE: To analyze the socioeconomic and gestational profiles, and the neuropsychomotor development of patients with infantile cataract cared for at a medical ophthalmologic center in Pernambuco state, Brazil. METHODS: A standardized questionnaire was applied, in October and November 2003, to the parents of children who underwent child cataract surgery at the Altino Ventura Foundation. RESULTS: Of the 40 patients, 23 were females (57.0%) and 17 males (43.0%). Most of the mothers (65.0%) and fathers (55.0%) of the children informed that they had not finished elementary school. The family income was between 1 and 3 minimum wages in 70.0% of the cases. As for the parental marital status, 40.0% of them were married. Thirty-nine mothers (97.5%) underwent prenatal exams, 37 reported that they had not received rubella immunization (92.5%), and 13 reported rubella during their pregnancy (32.5%). Three mothers reported consanguinity with their husbands and 5 were exposed to medications during pregnancy. Most of the patients had a neuropsychomotor development within limits considered normal by other studies. Fourteen patients (35.0%) were 6 years or older when the interview was performed. Eleven (78.6%) of these children attended school and 45.4% of them had to repeat the school grade. CONCLUSION: It was observed in the studied sample that the patients were characterized by being part of a low-income population. Less than half of the interviewed parents were married and the patients presented low school performance. Insufficiencies in the health system as regards immunization against rubella was identified. PMID- 15824798 TI - [Myotonic dystrophy type 1 in cataract patients: molecular diagnosis for screening and genetic counseling]. AB - PURPOSE: To detect MD1 premutation and full mutation carriers among cataract patients and offer familial genetic counseling. METHODS: We studied the DNA of 60 selected cataract patients through polymerase chain reaction analysis. This study was performed at the "Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto" where selected patients had been examined at the Cataract Outpatient Clinic from 01/01/1982 to 30/06/1995. Selection criteria were age under 55 with no obvious precipitating factor, except diabetes mellitus type 2, with or without neuromuscular signs suggestive of myotonic dystrophy. RESULTS: Three patients were found to have a full mutation corresponding to 5% of the group. Additional affected individuals were found among patients' relatives. No premutation was found. CONCLUSIONS: These results emphazise the importance of screening for MD1 gene carriers among cataract patients, and further genetic counselling. PMID- 15824799 TI - [Laboratory study of the mycotic infectious eye diseases and factors associated with keratitis]. AB - PURPOSE: To analyze the frequency and the etiology of mycotic infectious eye diseases diagnosed by culture at the Ocular Diseases Laboratory of the Federal University of Sao Paulo (LOFT-UNIFESP), Sao Paulo, SP, Brazil, and the factors associated with fungal keratitis. METHODS: We retrospectively analyzed the 14,391 laboratory requests regarding infectious eye diseases between 1975 and May 2003. For the fungal culture proven examinations we observed the morphologic type of the isolated fungus, the gender and age of the patient, the site of ocular involvement, the year of identification, and the factors associated with keratitis cases, according to the identified etiologic agent. Results were considered significant if p<0.05. RESULTS: We verified 296 (2.05%) ocular mycosis cases with 265 keratitis (89.5%), 27 intraocular infections (9.1%), and 4 infections at other sites (1.3%). Filamentous fungi were isolated in 233 cases (78.7%) 74.7% being male (n=174) and 25.3% female (n=59). Yeasts were isolated in 63 cases (21.3%) 49.2% being male (n=31) and 50.8% female (n=32). Ages varied between 2 and 99 years in the filamentous fungi group and between 14 and 88 years in the yeast group. The most frequently found etiologic agents of the filamentous fungi isolations were Fusarium (n=137; 58.8%), Aspergillus (n=28; 12.0%), and Penicillium (n=12; 5.2%); and in yeasts, Candida (n=59; 93.7%). Regarding keratitis, 95.1% had at least one associated factor. Ocular trauma was significantly associated with filamentous fungi keratitis (p=0.0002); and previous surgeries (p=0.0002), ocular (p=0.0002) and systemic diseases (p=0.0002) with yeast keratitis. The use of topic antibiotics showed association with filamentous fungi keratitis, at a lower statistical significance (p=0.0224). CONCLUSIONS: Laboratory analysis showed predominance of filamentous fungi over the yeasts, and keratitis was the main observed ocular infection. Fungal keratitis involved at least one associated factor in most cases. PMID- 15824800 TI - Comparison of PMMA, foldable silicone and foldable acrylic hydrophobic intraocular lenses in combined phacoemulsification and trabeculectomy. AB - PURPOSE: To compare the postoperative results of phacotrabeculectomy with implantation of PMMA, foldable silicone or foldable hydrofobic acrylic intraocular lens (IOL). SETTING: Glaucoma unit, The Royal Liverpool University Hospital, Liverpool, United Kingdom. METHODS: We studied a total of 124 eyes of three consecutive groups of patients with glaucoma and cataract that underwent phacotrabeculectomy with implantation of a PMMA (30 eyes), a foldable silicone (57 eyes) or a foldable acrylic (37 eyes) IOL. Postoperative Snellen visual acuity and intraocular pressure (IOP), and early and late complications were assessed. All data were analyzed by means of chi2 test, Fisher's exact test, ANOVA/MANOVA tests or a combination whenever appropriate. RESULTS: In all three groups the early and late mean postoperative IOPs were significantly lower than the preoperative ones (p<0.001), with no intergroup differences (p=0.48). The number of eyes with early postoperative hypertension (IOP>25 mmHg) and hypotony (IOP<7 mmHg) was similar in the three groups (p=0.91 and p=0.92 respectively). All groups showed improvement in mean visual acuity (p<0.001), and the differences among the groups were not significant (p=0.79). By 9-12 months after surgery IOPs lower than 22 mmHg without glaucoma medication were found in 76.9% in the PMMA group, 76.6% in the silicone group and in 76.9% in the acrylic group. At the same interval, best visual acuity of 6/12 or better was attained in 80.8%, 83% and 80.8%, in the PMMA, silicone and acrylic groups respectively. The silicone group had significantly more postoperative fibrin reaction into the anterior chamber (p=0.01) and giant cell deposits on the IOL (p<0.0001) than the PMMA and the acrylic groups. The rate of Yag laser posterior capsulotomy was lower with the acrylic IOL (0%) than with the silicone (12.2%) or PMMA (13.3%) IOLs (p=0.08). CONCLUSION: In patients with glaucoma and cataract, phacotrabeculectomy with PMMA, silicone or acrylic IOL was equally effective in lowering the IOP and improving visual acuity. However, incidence of fibrin reaction and lens deposits was higher in those eyes which received a silicone IOL. The PMMA and the acrylic groups did not differ with respect to postoperative complications, but those eyes with an acrylic IOL had a lower rate of posterior capsule opacification. PMID- 15824801 TI - [Cost-benefit analysis of ocular examination of patients with rosacea]. AB - PURPOSE: To observe the main ocular manifestations of patients with rosacea and to evaluate the cost-benefit of supplementary diagnostic examinations. METHODS: Twenty patients with acne rosacea from the Dermatology Service of the Sao Paulo Federal University (UNIFESP) were interviewed through a questionnaire, submitted to ophthalmologic examination and tested with Schirmer I, fluorescein and rose Bengal staining, impression cytology and conjunctival scrub. The costs of these examinations were abstracted from the Public Health System's values (SUS), Brazilian Medical Association values (AMB) and values charged by a private service. RESULTS: Twenty patients were enrolled initially in the study, but only 16 completed it. Eleven patients were female (55%), 19 were white (95%) and the average age was 56 years. All patients had at least one complaint related to rosacea. The most common symptoms were itching (40%), tearing (35%), foreign body sensation (30%). The most common signs were telangiectasis of lid margins (70%), hyperemia of lid margins (70%), meibomitis (65%) and blepharitis (50%). Most subsidiary examination showed normal results. There was difference of costs between the different paying sources. CONCLUSIONS: The main ocular manifestations in rosacea were related with meibomian gland disease. Considering the subsidiary examination costs and their results, the ophthalmologic check-up presents the best cost-benefit for the patient with rosacea. PMID- 15824802 TI - [Comparison between the pain induced by phacoemulsification with topical anesthesia and by peribulbar anesthesia without sedation]. AB - PURPOSE: To compare pain sensation induced by phacoemulsification with topical anesthesia with that by peribulbar anesthesia, without sedation. METHODS: Using a 10-level visual pain analogue scale, the pain induced by phacoemulsification with topical anesthesia (2% tetracaine drops) was measured in 20 patients. The same scale was used to measure the pain induced by peribulbar anesthesia (2% lidocaine and 0.5% bupivacaine) in 21 patients. The same surgeon performed peribulbar infiltrations and surgeries. The phacoemulsifications were done using clear corneal approach followed by in-the-bag intraocular lens implantation. Pain comparison between the two groups was performed by the non-parametrical U Mann Whitney test. RESULTS: The distribution of the pain scores of the patients who underwent cataract extraction with topical anesthesia ranged from 0 to 5 (median value = 2). The scores of the patients who underwent peribulbar infiltration ranged from 0 to 7 (median value = 3). The mean rank of the group of patients who had surgery with topical anesthesia (15.78) was significantly different from the mean rank of the patients who received peribulbar anesthesia (25.98) (p = 0.0057). CONCLUSION: Without sedation, the pain induced by phacoemulsification with topical anesthesia was milder than the pain induced by peribulbar infiltration. PMID- 15824803 TI - [The role of frequency doubling perimetry in the diagnosis of glaucoma: screening in employees of a public hospital in an urban area of Sao Paulo]. AB - PURPOSE: To study the prevalence of glaucoma in employees of a public hospital in Sao Paulo and to analyze the role of frequency doubling perimetry along with non contact tonometry and direct ophthalmoscopy, in the screening for glaucoma. METHODS: 612 employees of the Central Hospital of the "Santa Casa de Sao Paulo" were evaluated from October 15 to 20, 2000; 438 were women and 174 men, with mean age of 45.05+/-7.7 years (range, 35 to 81 years); 437 were white, 104 mulatto, 43 black and 28 Asian. All examinations were done by 10 eye specialists with experience in glaucoma. All employees had both eyes examined, by means of frequency doubling perimetry (FDT), ophthalmoscopy (FO), and non-contact tonometry (TNC). RESULTS: 159 (25.98%) subjects presented with at least one abnormal testing; 5 (3.14%) had high IOP (TNC+); 13 (8.17%) presented suspicious disk (FO+); 110 (69.18%) presented abnormal FDT (FDP+); 8 (5.03%) TNC+ and FO+; 10 (6.28%) had TNC+ and FDT+; 9 (5.66%) presented FO+ and FDT+; 4 (2.51%) had TNC+, FO+ and FDT+; 12 (1.96%) individuals were diagnosed with glaucoma, four of whom (0.65%), with normal pressure glaucoma. CONCLUSIONS: Frequency doubling perimetry was found to be an important step in the screening for glaucoma in the study. Of the 12 individuals that had the diagnosis of glaucoma, 5 (41.6%) would not have been diagnosed if they had not undergone frequency doubling perimetry. PMID- 15824804 TI - [Cost of phacoemulsification in the national campaign of elective cataract surgery in Itapolis, SP, Brazil]. AB - PURPOSE: To determine the average cost of cataract surgery using the phacoemulsification technique with intraocular lens implant (IOL). This study was conducted during a national campaign to treat patients with cataracts promoted by the Brazilian Council of Ophthalmology (CBO) and the Health Ministry in the city of Itapolis-SP from March to December, 2000. METHODS: All expenses related to the surgery were collected (pre-, intra-, and postoperative data) and analyzed. Fifty eight patients with senile cataracts and without any other ocular findings were submitted to the procedure in a prospective study. RESULTS: The average cost of the surgery in this study was R$ 485.03 or USD 248.05. This value represents the average cost of the surgery itself; in which some materials, equipment, taxes and other services were previously determined and used for this project. CONCLUSION: The phacoemulsification technique for cataract surgery has been used more and more, due to its effectiveness, being an excellent method to treat cataracts. Although this is an excellent technique, the economical aspect does not allow it to be used for part of the population. PMID- 15824805 TI - [Strabismus surgical treatment: technical-economic evaluation]. AB - PURPOSE: To evaluate, from the economic and technical point of view, the surgical treatment of strabismus. METHODS: A retrospective survey of available consecutive medical charts from the Altino Ventura Foundation-Recife/PE, Brazil was performed. Data on 100 patients who underwent strabismus surgery (from January of 2001 to February of 2003) and 100 patients who underwent cataract surgery (January of 2003) were analyzed. The times to performed these surgical procedures were observed. The times to accomplish strabismus surgeries were compared to those of cataract surgeries. In addition the variability of time among different strabismus surgeries was also observed. RESULTS: Surgical mean time for strabismus correction was longer than that for cataract surgery (77.3+/-27.6 min versus 58.3+/-12.7 min; p<0.0001). A significant variability in time to accomplish strabismus surgeries was also observed. In the case of intervention in up to two muscles, mean time was less than in the case of intervention in more than two muscles (66.0+/-16.6 min versus 97.5+/-31.7 min; p<0.0001). CONCLUSION: Longer surgical time of strabismus surgeries as compared to cataract surgeries gives support to the idea that the medical remuneration for strabismus surgery should be, at least, equivalent to that of cataract surgery. In addition, the fact of surgical mean time being so variable, suggests that this procedure should have a different remuneration. PMID- 15824806 TI - [Reliability of corneal thickness measurements by Orbscan II and ultrasound pachymeter]. AB - PURPOSE: To access the reliability of corneal thickness measurements by Orbscan II and ultrasound pachymeter. METHODS: A retrospective study of 134 eyes from 67 normal subjects between January and June 2001 was obtained. Sex, age, visual acuity, refractive error and corneal thickness measurements by Orbscan II and ultrasound pachymetry were evaluated. All results were analyzed by the paired t test (p<0.05). RESULTS: Sixty-seven subjects, 34 (50.7%) female and 33 (49.3%) male, with a mean age of 32.44+/-9.98 were evaluated. The mean of spherical equivalent was -2.68+/-2.62. The mean corneal thickness was 534.81+/-34.45 with the Orbscan II system and 535.00+/-29.53 with the ultrasound pachymeter, values that were not significantly different (p=0.8922). The correlation coefficient between both instruments was 0.8774, and it is effective. CONCLUSIONS: Both methods are similar showing good correlation. In this study, the Orbscan II showed reliability, and this tool is extremely useful to evaluate subjects regarding refractive surgery. PMID- 15824807 TI - Epidemiologic study of anisometropia in students of Natal, Brazil. AB - PURPOSE: To perform an epidemiologic study in students in Natal/Brazil, with relation to refractional anisometropia, evaluating criteria such as: gender, age, and association with strabismus and amblyopia. METHODS: A study of 1,024 students randomly selected from several districts of Natal/Brazil was undertaken by the Department of Ophthalmology of the Federal University of Rio Grande do Norte (UFRN), observing the following criteria of > or =2 spherical or cylindrical diopter refractional anisometropia relating it to sex, age, association with strabismus, amblyopia and anisometropia classification. RESULTS: We found a prevalence of 2% (N=21) anisometropia in the students. The female gender predominated with 81% (N=17). In students with anisometropia, we observed an association with strabismus in 9.5% of cases (N=2), both with exotropia. The association of anisometropia with amblyopia occurred in 47.6% of the cases (N=10), with 8 cases of unilateral amblyopia and 2 cases of bilateral amblyopia. CONCLUSIONS: There was a predominance of anisometropia in females, and an increased prevalence of strabismus and amblyopia in students with anisometropia. PMID- 15824808 TI - [Economic impact of eyedrop cost in glaucoma treatment]. AB - PURPOSE: To evaluate the daily cost of antiglaucoma eyedrops and the economic impact related to the minimal wage; to compare the cost of drug association in the single presentation in relation to separate presentations; to analyze the additional percent antiglaucoma drug cost related to standard therapy (generic timolol maleate). METHODS: Fifteen eyedrop bottles of each one of the twenty antiglaucoma products were used. The number and mean eyedrop size per bottle of drug were measured and the duration and treatment costs calculated. RESULTS: A large variation in the mean daily cost, respectively: R$ 0.077 for generic timolol maleate (cheapest) and R$ 1.910 for Xalacom (most expensive), was observed. It should be noted that the product of minimal economic impact, as related to the minimal wage, was the standard eyedrop therapy with 1.2% to 1.6%, while the association of Xalatan with Timoptol XE showed a variation of 21.7% to 30.0%. The cost of Cosopt and Xalacom was greater than the associations of, respectively: Trusopt + standard eyedrop and Xalatan + standard eyedrop (p<0.001). Xalacom represented an additional cost to standard therapy of 1.698.2% to 1.765.1%. CONCLUSIONS: The variation of antiglaucoma eyedrop cost was almost 30 times between the cheapest and the most expensive, which represented an economic monthly impact of 29.1% on the value of the minimal wage. The combined therapy in the separate presentations presented a lower cost than drug association in a single presentation. The use of antiglaucoma drugs represents a high percent additional cost related to standard therapy. PMID- 15824809 TI - [Eyelid measures before and after upper blepharoplasty--quantitative evaluation]. AB - PURPOSE: To evaluate eyelid measures before and after upper blepharoplasty surgery using a digital image system. METHODS: Eighteen eyelids of 9 patients with dermatochalasis were evaluated at the Hospital das Clinicas de Botucatu UNESP regarding eyelid measures before and after upper blepharoplasty. All patients were female, aged between 40 and 75 years. The images were acquired by a photographic camera, transferred to a computer and then analyzed by the Scion Image Frame Grabber program. The evaluated parameters were upper eyelid position in primary gaze position, upper palpebral sulcus and lateral eyelid angle. The measures were obtained before and after 60 days of upper dermatochalasis surgery. RESULTS: After the surgery, there was an increase of upper eyelid position and upper palpebral sulcus; the lateral palpebral angle did not show any alteration. CONCLUSION: Eyelid position changes after blepharoplasty and digital image processing provides quantitative measures of the eyelid, allowing a better quantification of the surgical results. PMID- 15824810 TI - [Dacryocystography in adult lacrimal system evaluation]. AB - PURPOSE: To evaluate dacrycystographic examination in patients with suspicion of lacrimal apparatus obstruction. METHODS: A retrospective study was performed to evaluate dacryocystographies of 100 adults suspected of nasolacrimal duct obstruction at the "Faculdade de Medicina de Botucatu-UNESP". The data were submitted to descriptive statistical analysis and chi-square test. RESULTS: The patients were mainly female, over 60 years old, presenting a low obstruction located at Arlt's sinus, with a grade 2 or 3 lacrimal sac. Lacrimal pathways with normal tear drainage, with or without dilatation and nasal alterations such as concha hypertrophy were also observed. CONCLUSION: Dacryocystographic evaluation was important to show the level of obstruction, the presence of dilatation of the lacrimal sac, as well as alterations in nearby structure. This information might influence the adopted surgical technique and the final treatment results. PMID- 15824811 TI - [Contact lens fitting after refractive surgery]. AB - PURPOSE: To evaluate the fitting and use of contact lens in patients submitted to refractive surgery. METHODS: This was a retrospective study in 53 patients submitted to refractive surgery who later started to use contact lens, from 1999 to 2003. The parameters were: previous ametropia, refractive surgery procedure, ametropia after surgery, postoperative spherical equivalent, postoperative keratometry, base curve of fitted contact lens, contact lens design, final visual acuity with spectacles after surgery, final visual acuity with contact lens, complications and the reason for stopping the use. The follow-up of patients ranged from 1 month to 84 months (average of 42.5 months). RESULTS: Of 53 evaluated patients, 19 patients had undergone LASIK (Laser Assisted in Situ Keratomileusis), 29 patients had undergone RK (radial keratotomy), 4 patients had undergone PRK (photorefractive keratectomy) and in one patient it was not possible to know which surgery was performed. 61.29% of the patients (57 eyes of a total of 93 eyes) were fitted with rigid gas-permeable lenses. There was an improvement of visual acuity in 60.21% of the cases (VA>20/40), with few complications. CONCLUSION: Due to the great number of performed refractive surgery, an increase in the number of patients unhappy with the postoperative result is expected, and for these patients, many times the use of contact lens is the best option. The fitting of contact lenses after refractive surgery demands knowledge, dedication and has good results principally regarding better visual acuity. PMID- 15824812 TI - [Conjunctival autograft transplantation in primary pterygium]. AB - PURPOSE: To evaluate the use of conjunctival autograft transplantation in the treatment of primary pterygium. METHODS: Prospective study of 230 patients (230 eyes), with primary pterygium, who underwent conjunctival autograft as a treatment option for recurrent pterygium. The minimal follow-up was 12 months. RESULTS: We observed a recurrency rate of 2.17%, 0.43% had loss of graft. CONCLUSION: Conjunctival transplant is a procedure that rarely brings any complication. However, the literature on the results leads to new clinical and randomized, studies, in order to evaluate the responses of the conjunctival autograft transplant as related to the other surgical techniques for pterygium. PMID- 15824813 TI - [Tarsorrhaphy: applications in a Cornea Service]. AB - PURPOSE: To evaluate the main indications for tarsorrhaphy in a Corneal Service, as well as success rate and possible complications. METHODS: All patients who underwent tarsorrhaphy from January 1st, 2002 to December 30th, 2002, in Sao Geraldo Eye Hospital - Federal University of Minas Gerais were evaluated retrospectively. Reviewed data included patient's sex and age, indication for tarsorrhaphy, duration of signs and symptoms before tarsorrhaphy, time for epithelial healing after tarsorrhaphy, type of tarsorrhaphy, complications, follow-up time. RESULTS: Eighteen patients underwent tarsorrhaphy. The indications for a tarsorrhaphy were exposure keratopathy (27.8%), persistent epithelial defect associated with penetrating keratoplasty (38.8%), neurotrophic ulcer (11.1%), dry eye syndrome (5.6%), Stevens-Johnson syndrome (11.1%), and chemical burn (5.6%). The epithelial defect resolved completely in 15 patients (83.3%). Mean duration of signs and symptoms before tarsorrhaphy was 98.7+/-48.6 days, and time to heal after tarsorrhaphy was 53.2+/-22.8 days. Of the 18 procedures, two (11.1%) were temporary and 16 (88.9%) permanent. Complications after tarsorrhaphy were premature opening of the tarsorrhaphy, trichiasis and pyogenic granuloma. CONCLUSION: Tarsorrhaphy is a simple procedure, being also very safe and effective in the management of nonhealing epithelial defects, with a success rate of 83.3% and with minor complications. PMID- 15824814 TI - [Comparative study of the density of corneal endothelial cells after phacoemulsification by the "divide and conquer" and "quick chop" techniques]. AB - PURPOSE: Since one of the most implicated factors in corneal endothelial cell loss after phacoemulsification is the ultrasound time, and that the "quick chop" technique utilizes less ultrasound time, the objective of this study was to compare and analyze the reduction of the central corneal endothelial cell density between the patients submitted to the "divide and conquer" and "quick chop" techniques. METHODS: Fifty-six patients with grade 3 senile cataract were prospectively selected and submitted to endocapsular phacoemulsification by the "divide and conquer" (28 patients, group 1) and "quick chop" (28 patients, group 2) techniques. The non-contact corneal central specular microscopy performed pre- and postoperatively at one month, 3 months and 6 months, and the ultrasound time used for each surgery, were verified. RESULTS: We observed that the "divide and conquer" technique spent an average of 2.0 minutes of ultrasound time and the "quick chop", 1.1 minute; that the reduction of the central corneal endothelial cell density was significant for both techniques in the first postoperative month (16.5% in group 1 and 19.4% in group 2) and that the variations in central corneal endothelial cell density that followed, at the 3rd and 6th month were not significant. There was no statistically significant difference between the variation of the central corneal endothelial cell density that occurred with both techniques (Student's "t" test p=0.334; in the 1st postoperative month). CONCLUSIONS: We concluded that the reduction of central corneal endothelial cell density was not statistically different between the two techniques. PMID- 15824815 TI - [Visual outcome with binocular implantation of SA-40N multifocal intraocular lens at "Hospital de Olhos de Minas Gerais"]. AB - PURPOSE: To evaluate the outcomes of phacoemulsification with binocular implantation of SA-40N multifocal intraocular lens at the "Hospital de Olhos de Minas Gerais". METHODS: The charts of 58 patients (116 eyes) who were submitted to phacoemulsification with binocular implantation of SA-40N multifocal intraocular lens were retrospectively analyzed. RESULTS: The mean preoperative spherical equivalent was +1.55D (ranging from -4.25 to +6.25D) and after surgery it was -0.59D (ranging from -2.25 to +1.25D). Thirty days after surgery, 94 eyes (81%) achieved 20/20 for better distance corrected visual acuity. For best uncorrected binocular near vision, 29 (50%) patients were able to read J1 and 25 (43.1%), J2. When we compare the best corrected distance vision, before and after surgery, we observed that 68 (58.6%) eyes keep the same best corrected visual acuity, 33 (28.4%) eyes gained lines of visual acuity and 15 (12.9%) lost lines of vision. Regarding complications, 19 (32.7%) reported halos, three (2.6%) eyes had macular edema and five (4.3%) eyes had posterior capsular opacification. Thirty-six patients (62.1%) did not need to use spectacles. CONCLUSION: The results of this study indicate that multifocal intraocular lens bring about less dependence on spectacle wear and high patient satisfaction despite some reports of halos. PMID- 15824816 TI - [Unilateral keratoconus associated with continual eye rubbing due to nasolacrimal obstruction--case report]. AB - To describe a possible case of unilateral keratoconus in a patient with continual eye rubbing due to an ipsilateral nasolacrimal obstruction in a 17-year-old woman complaint of progressing low visual acuity in the left eye. Presented with constant epiphora in the left eye throughout her life as a result of ipsilateral nasolacrimal obstruction, which required frequent wiping of this eye. Based on ocular examination, the patient was diagnosed as having unilateral keratoconus in the left eye. As in other studies, keratoconus is thought to be associated with persistent eye rubbing. This patient represents a case of unilateral keratoconus, where there is a confirmed history of habitual eye rubbing due to ipsilateral nasolacrimal obstruction, thereby suggesting a possible causal relationship between eye rubbing and keratoconus. PMID- 15824817 TI - [Bilateral central retinal vein occlusion associated with blood hyperviscosity syndrome--case report]. AB - The authors report the case of a 16-year-old male patient who presented with blurred vision and bilateral optic disc edema, then developing bilateral central retinal vein occlusion. On laboratory work-up, he was found to have multiple myeloma IgA along with hyperviscosity syndrome, which led to the ophthalmological features. After proper treatment, the patient recovered visual acuity and normalized his eye fundus changes. Bilateral central retinal vein occlusion finding may yield the diagnosis of major systemic diseases. Fundoscopic features may serve as parameters on treatment evaluation. PMID- 15824818 TI - [Charles Bonnet syndrome: visual hallucinations in patients with ocular diseases- case report]. AB - In this article the authors report two cases of Charles Bonnet syndrome, defined as complex visual hallucinations in patients with low vision, and the patient is aware of the unreal nature of the phenomenon. A great number of cases is misdiagnosed due to lack of direct questioning by the physician. Since the emotional distress caused by this disease, the knowledge of its symptoms is essential in the management of these patients. PMID- 15824819 TI - [Ocular aspergillosis--case report]. AB - Presence of fungus in the conjunctiva is a constant threat to the eyes, because these microorganisms, defined as opportunistic, may provoke severe ocular infections, in situations as low organic resistance, use of immunosuppressants, antibiotics and epithelial alteration. Our goal here is to report a case of ocular aspergillosis in an immunosuppressed patient where paroxysmal nocturnal hemoglobinuria was diagnosed. A 51-year-old immunosuppressed, thrombocytopenic patient was hospitalized with a diagnosis of paroxysmal nocturnal hemoglobinuria. On examination, she had a visual acuity of 20/40 in the right eye (RE) and light perception in the left (LE). She presented hyposphagma, bilateral conjunctival edema on biomicroscopy and had some multiple and circumscribed conjunctival abscesses in both eyes; clear cornea in both eyes. Fundoscopy of the RE did show any particularity, in the LE there was a smear hemorrhage. Computed tomograph showed a fat periocular infiltration. Magnetic resonance lead to the same finding, compatible with orbital cellulitis. Hemoculture showed Aspergillus growth and direct smears of conjunctival material was negative. There was great improvement while treating her with amphotericin B, but there was complete remission after using 5% natamicyn eye drops. Orbital infections caused by Aspergillus are uncommon, usually appearing in immunosuppressed patients. Frequently they progress insidiously, and can be confounded with other orbital processes. Immunological impairment can inhibit the expression of local and systemic symptoms, resulting in diagnostic confusion. The diagnosis is established by laboratory tests, but culture can be negative in spite of the classical presentation, making the beginning of treatment difficult. In these cases management starts according to the symptoms. PMID- 15824820 TI - [Eyelid trichoepithelioma--report of 2 cases]. AB - Trichoepithelioma is a benign skin tumor and is most commonly found on the face, however, there are few reports about its occurrence on the eyelids. It is not usually recognized because of its rarity, origin, biological potential, and controversial classification. The purpose of this paper is to report two cases of solitary trichoepithelioma on the eyelid, their histopathology and treatment. This lesion should be considered when a single solid nodule on the face appears, and can be differentiated from basal-cell carcinoma. Histopathologic confirmation is essential. PMID- 15824821 TI - [Corneal wound healing response following different modalities of refractive surgical procedures]. AB - The corneal wound healing response following refractive procedures represents a subject of high relevance, due to its direct influence on the postoperative results. Technical modifications of current refractive procedures, like the automated flap creation with the femtosecond laser, LASEK, PRK with mitomycin C and Epi-LASIK have been proposed as alternatives to traditional LASIK and PRK. Several theoretical advantages have encouraged the diffusion of these new techniques; however, a better understanding of the corneal wound healing response following these procedures is required. The present text proposes a review of the corneal wound healing characteristics following different modalities of refractive surgical procedures. PMID- 15824822 TI - [Sad orbit, or how dissimilar . . ]. PMID- 15824823 TI - [Relationship of oncogene C-erbB-2 amplification in breast cancer with pathological parameters and disease free survival]. AB - BACKGROUND: Proto-oncogene c-erbB-2 is located in chromosome 17 region q21 and codifies a 185 Kd protein, with tyrosine kinase activity. The amplification of this gene is associated with relapse and lower survival in breast cancer. Overexpression of this gene can be detected by immunohistochemistry (IHC). However, fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) allow the simultaneous analysis of morphology and overexpression of the gene. AIM: To evaluate the relationship of c-erbB-2 oncogene amplification measured by FISH with histological graduation, presence of positive Iymph nodes and evolution of breast cancer. PATIENTS AND METHODS: One hundred and ten tissue samples of invasive ductal or lobulillar breast cancer, positive for c-erbB-2 oncogene by IHC were analysed. The presence of c-erbB-2 oncogene amplification was subsequently analyzed by FISH. RESULTS: There was a significant association of c-erbB-2 amplification by FISH with pathological graduation of the tumor, number of regional Iymph nodes involved and disease free survival. CONCLUSIONS: Proto-oncogene c-erbB-2 amplification is a good indicator of bad prognosis in invasive breast cancer. PMID- 15824824 TI - [Results of radiofrequency ablation of the inferior vena cava-tricuspid valve isthmus for atrial flutter]. AB - BACKGROUND: Radiofrequency ablation of the inferior vena cava-tricuspid valve isthmus relieves atrial flutter in 95% of cases. AIM: To evaluate the long term results of radiofrequency ablation of the inferior vena cava-tricuspid valve isthmus in atrial flutter. MATERIAL AND METHODS: Retrospective review of 86 patients with common atrial flutter, treated with radiofrequency ablation of the isthmus, while in sinus rhythm or flutter. Patients were contacted by telephone for a new clinical and electrocardiographic assessment, three to 40 months after the procedure. RESULTS: Of all the patients treated, five died and five were lost from follow up, thus 76 patients (mean age 56 years, 58 males) were contacted for reassessment. At the moment of the procedure, 51% had an underlying cardiac disease and 25% had high blood pressure. All referred palpitations, 25% had dyspnea, 84% were receiving antiarrhythmic drugs and 33% were on oral anticoagulants. Flutter was paroxystic in 83% and chronic in 17%. Fulguration was successful in all patients; one patient presented a high grade atrioventricular block as a complication of the procedure. At reassessment, 82% of patients were in sinus rhythm, 16% had atrial fibrillation and 2%, an atypical flutter. CONCLUSIONS: Radiofrequency fulguration is a safe and effective treatment of atrial flutter. PMID- 15824825 TI - [Markers of thrombophilia in patients with retinal vein thrombosis]. AB - BACKGROUND: Thrombophilia is an alteration of hemostasis that increases the risk to venous or arterial thrombosis. This condition may be the underlying cause of retinal vein thrombosis. AIM: To study the presence of thrombophilia in patients with retinal vein thrombosis. PATIENTS AND METHODS: Prospective study of 55 patients aged 22 to 86 years, with retinal vein thrombosis (central or branch). Antithrombin III, coagulant protein C, functional protein S, resistance to activated C protein, homocysteine, prothrombin G20210A gene, lupus anticoagulant and anticardiolipin antibodies were measured in all. RESULTS: Seventeen patients had thrombophilic markers (antiphospholipid syndrome in seven, hyperhomocysteinemia in six and resistance to protein C in three). Of these 17 patients, 53% had high blood pressure, 35% an abnormal serum lipid profile and 23% a personal history of thrombosis. The thrombosis was central in 12 (ischemic in four) and of a branch in five (ischemic in two). CONCLUSIONS: Thrombophilic markers must be assessed in patients with retinal vein thrombosis. PMID- 15824826 TI - [Proposal of a method to assess global quality of diet]. AB - BACKGROUND: Several studies have attempted to assess the global quality of diet. One of such initiatives is the Healthy Eating index (HEI), developed in the United States. However, its application in other countries requires certain adaptations. AIM: To adapt the HEI to the Chilean population and to assess its value in Chilean school age children and adults. MATERIAL AND METHODS: A random sample of 264 school age children aged 13.4 +/- 0.7 years and 264 adults aged 35.7 +/- 0.6 years was studied. A 24 hours food recall was done and the HEI seas calculated considering 10 variables based in the Feeding Guides for the Chilean population. Feeding habits were classified as healthy when the score was over 80 (of a maximum of 100), as requiring changes when the score was between 51 and 80 and as unhealthy when the score was 50 or less. RESULTS: The mean scores for children and adults were 58.4 +/- 11.3 and 56.2 +/- 11.1 respectively (p< 0.02). The variables with lower scores were vegetables, fruits, dairy products and sodium. Only 1.5% of the population had healthy feeding habits, without differences by sex or age. Feeding quality was significantly worse among obese subjects. CONCLUSIONS: The HEI is a tool that can be used in the Chilean population and can be used to assess the effectiveness of health promotion initiatives. It also can help to focalize educational programs to specific nutrients. PMID- 15824827 TI - [Periodontal disease among patients with acute coronary syndrome]. AB - BACKGROUND: Epidemiological studies suggest an association between periodontal disease and coronary heart disease. It is possible that periodontal disease may contribute to plaque destabilization in patients with acute coronary syndrome. AIM: To assess the association between severity of periodontal disease, the number of acute plaques and extension of coronary artery disease in patients with acute coronary syndrome. PATIENTS AND METHODS: The severity of periodontal disease was assessed by skilled independent observers in patients with acute coronary syndrome subjected to coronary angiography. The periodental disease was classified as mild or severe. Acute plaques were defined on angiography as those with thrombi, ulcers or flow alterations. The extension of coronary disease was analyzed using the Sullivan score. RESULTS: Forty three patients (35 males) aged 41 to 83 years, were studied. Mild and severe periodontal disease was present in 18 (42%) and 25 (58%) patients respectively. Seventy six percent of those with severe disease had two or more acute plaques, compared with 17% of those with mild disease (p<0.001). Median Sullivan score was 80.6 and 57.2 in patients with severe or mild periodontal disease respectively (p=0.001). CONCLUSIONS: Severe periodontal disease was associated with a higher number of acute coronary plasques and a higher extension of coronary artery disease, in patients with acute coronary syndromes. PMID- 15824828 TI - [Survival of patients with brain metastases]. AB - BACKGROUND: Brain metastases are the most common cerebral tumors, have a poor prognosis and their incidence is five times higher than primary brain tumors. AIM: To analyze the survival of patients with the diagnosis of brain metastases, operated in our institution. PATIENTS AND METHODS: We retrospectively reviewed all patients operated from January 1989 to December 2001, whose pathological diagnosis confirmed the presence of cerebral metastases. The death date of each patient was obtained from the analysis of death certificates, obtained from the computerized National Registry. RESULTS: In 46 operated patients, the date of death was determined. In 23 of them, information about primary site of malignancy, type of surgery performed and adjunctive treatment with Holocerebral Radiotherapy (Rt) was obtained. The overall median survival time of the 46 patients was 29 weeks (range 4-207). Thirty percent of patients were alive one year after surgery. Among those patients with complete clinical information, the median survival of 16 patients who received postoperative Rt was 41 weeks (range 12-207), compared to a survival of 18 weeks (range 8-72), among those that did not receive Rt (p = 0.04). CONCLUSIONS: The median survival for patients operated for cerebral metastases in our institution is 29 weeks. Those who are operated and receive Rt, have a longer survival, than those who did not receive Rt. PMID- 15824829 TI - [A proposal of new indices for hospital management]. AB - BACKGROUND: Diagnosis related groups (DRGs) are the most reliable patient classification system in hospital management. When this information is unavailable, other reliable classification system must be used. AIM: To obtain useful indices for hospital management, based on descriptive multivariate techniques. MATERIAL AND METHODS: Data on admissions to a University Hospital during 2003 were analyzed. Number of discharges, lethality rate, re-admission rate, number of outpatient consultations, length of hospital stay and surgical complexity index were analyzed, using information obtained by the Operations Management Department. The Principal Components Analysis (PCA) technique was applied and the R correlation matrix was used. RESULTS: A total of 24,345 discharges were analyzed. The first two principal components were selected, accounting cumulatively for 76% of data variability (47% for the first and 29% for the second). CONCLUSIONS: The first component may be assimilated to a new index representing the difficulty of the attended cases, which we have termed Case Complexity. The second principal component would explain the number of attended persons, which we have termed Case Load. These two indices allow us to classify hospital services. PMID- 15824830 TI - [Skin prick test of immediate hypersensitivity in a selected Chilean pediatric population sample]. AB - BACKGROUND: Skin prick test (SPT) of immediate hypersensitivity is a main instrument in the diagnosis of allergy. AIM: To demonstrate the applicability of skin prick test in different age groups. PATIENTS AND METHODS: We studied children and adolescents with the diagnosis of allergy in the Pediatric Respiratory Laboratory of the Catholic University of Chile, from January 2001 to March 2002. The SPT was performed using a standardized technique. The allergens were applied on the volar surface of the forearm in children older than 4 years of age and in younger children it was applied on their back. For study purposes we separated them into three age groups: GI < or =2 years and 11 months, GII from 3 to 4 years and 11 months, GIII > or =5 years. RESULTS: We studied 408 children, aged between 8 months and 15 years. The SPT was applied to all patients with no adverse effects of any kind. There was a positive reaction in 57.7% of children. The reaction was positive in 37% in G1, 39% in GII and 65% in GIII (p <0.001). The predominant allergens for each group were dust mites (Dermatophagoides pteronissinus and farinae). CONCLUSIONS: SPT was useful when used on a selected pediatric population. The frequency of sensitization increased significantly with age. However, more than one third of children between 2 and 4 years of age tested positive to one or more allergen, demonstrating its applicability in this age group. PMID- 15824831 TI - Transverse myelitis and chronic urticaria in systemic lupus erythematosus. Case report. AB - We report a 40 years old woman with chronic urticaria and acute transverse myelitis associated with systemic lupus erythematosus. The urticaria appeared in her adolescence and after 26 years was followed by photosensitivity, peripheral polyarthritis and acute transverse myelitis, with positive antiphospholipid and antinuclear antibodies. Both chronic urticaria and acute transverse myelitis have been described associated with or appearing as the first manifestation of systemic lupus erythematosus. Transverse myelitis is a rare and still poorly understood condition reported in about 2% of patients with systemic lupus. PMID- 15824832 TI - [Pure autonomic failure. Bradbury Eggleston Syndrome. Case report]. AB - Pure Autonomic Failure is a progressive, adult onset, degenerative disorder of the autonomic nervous system characterized clinically by orthostatic hypotension, bladder, sexual and sudomotor dysfunction. Since there are no other associated somatic neurological deficits, this condition must be considered in the differential diagnosis of orthostatic hypotension. We report a 64 years old man with a history of seven years of autonomic dysfunction, with severe orthostatic hypotension, erectile and bladder dysfunction. Autonomic tests showed low circulating norepinephrine levels, sweating abnormalities with regional anhydrosis of the left side of the trunk and abnormal cardiovagal response, indicating generalized autonomic failure. Peripheral somatic neuropathies with autonomic involvement were excluded by normal electrophysiologic tests and the patient was diagnosed pure autonomic failure. Treatment with fludrocortisone and midodrine improved orthostatic tolerance. PMID- 15824833 TI - [Severe loxoscelism with lethal outcome. Report of one case]. AB - Loxoscelism, is caused by the bite of Loxosceles laeta spiders. It has two clinical forms: cutaneous loxoscelism (CL) and viscerocutaneous loxoscelism (VCL). VCL is characterized by hematuria, hemoglobinuria, jaundice, fever and sensorial involvement. In severe cases there is massive hemolysis and renal failure, with high lethality. We report a 71 year-old man, brought to the hospital five days after suffering a spider bite. The patient was admitted with a severe kidney failure, hemolysis, metabolic acidosis and clotting disorder. The patient was managed with adrenal steroids and dialisys but died five weeks after hospital admission. This particular patient consulted late and had multiple factors of poor prognosis. PMID- 15824834 TI - [An update on the pharmacological treatment of Alzheimer disease]. AB - Dementia in general, especially Alzheimer disease and vascular dementia, are diseases of high prevalence with severe socio-economic consequences in all countries. In recent years, due to the obtention of new pharmaceutical products acting on different brain neurotransmitters, there has been important changes in the therapy of these diseases. Although these drugs do not stop disease progression, there is consistent evidence of their usefulness in cognitive, behavioral and functional domains and of their pharmaco-economical justification. This article reviews the main drugs available for Alzheimer disease and some future therapeutic perspectives. PMID- 15824835 TI - [Evolutionary medicine: an emergent basic science]. AB - Evolutionary Medicine is an emergent basic science that offers new and varied perspectives to the comprehension of human health. The application of classic evolutionary theories (descent with modification, and natural selection) to the human organism, to its pathogens, and their mutual co-evolution, provides new explanations about why we get sick, how we can prevent this, and how we can heal. Medicine has focused mainly on the proximate or immediate causes of diseases and the treatment of symptoms, and very little on its evolutionary or mediate causes. For instance, the present human genome and phenotypes are essentially paleolithic ones: they are not adapted to modern life style, thus favoring the so-called diseases of civilization (ie: ateroesclerosis, senescence, myopia, phobias, panic attacks, stress, reproductive cancers). With the evolutionary approach, post modern medicine is detecting better the vulnerabilities, restrictions, biases, adaptations and maladaptations of human body, its actual diseases, and its preventions. PMID- 15824836 TI - [Experience with electronic files in a university neonatology unit]. AB - The electronic file is a reality in medical practice nowadays. We have a decade of experience with electronic files in a neonatology unit. We use a local network that consists in one server and 8 connected computers, distributed in the hospital. Filemaker Pro is used as database administrator and access to data is protected with passwords. Data entry is made by health care professionals in charge of the patients. Patient's reports and statistical information are based on data entered to the system. This methodology allows to have update clinical data, indexing of information, to maintain track of pharmacological indications, prescribe parenteral nutrition and obtain information for research purpose. It is possible therefore, with a minimal computing expertise, to devise electronic files that can improve the quality of health care. PMID- 15824837 TI - [Systematic reviews and metaanalysis: are the best evidence?]. PMID- 15824838 TI - [Critical appraisal: enteral nutrition is better than parenteral nutrition for patients with acute pancreatitis]. AB - OBJECTIVE: To compare the safety and clinical outcomes of enteral and parenteral nutrition in patients with acute pancreatitis. DATA SOURCES: Medline, Embase, Cochrane controlled trials register, and citation review of relevant primary and review articles. STUDY SELECTION: Randomized controlled studies that compared enteral nutrition with parenteral nutrition in patients with acute pancreatitis. From 117 articles screened, six were identified as randomized controlled trials and were included for data extraction. DATA EXTRACTION: Six studies with 263 participants were analyzed. Descriptive and outcome data were extracted. Main outcome measures were infections, complications other than infections, operative interventions, length of hospital stay, and mortality. The metaanalysis was performed with the random effects model. DATA SYNTHESIS: Enteral nutrition was associated with a significantly lower incidence of infections (relative risk 0.45; 95% confidence interval 0.26 to 0.78, P= 0.004), reduced surgical interventions to control pancreatitis (0.48, 0.22 to 1.0, P= 0.05), and a reduced length of hospital stay (mean reduction 2.9 days, 1.6 days to 4.3 days, P <0.001). There were no significant differences in mortality (relative risk 0.66, 0.32 to 1.37, P = 0.3) or non-infectious complications (0.61, 0.31 to 1.22, P= 0.16) between the two groups of patients. CONCLUSIONS: Enteral nutrition should be the preferred route of nutritional support in patients with acute pancreatitis. PMID- 15824839 TI - [Physicians and professionalism. Patients and information]. AB - In the present time, the advances in sciences brought by technology are worshiped. This contributes to dehumanize human relationships. Medical profession is a commitment to oneself, society and social justice. Scientific and humanist formation must coexist during the training of new physicians. The main objective of medical profession is the relationship with patients. Patients have acquired increasing autonomy and a deliberative relationship is the closest to the ideal medical action. The relationship must be based in autonomy, beneficence, lack of maleficence and justice. The patient must receive all the information about his ailment and the diagnostic and therapeutic alternatives in a clear and intelligible language. This will allow to obtain a fully conscious informed consent from the patient. Asking for this consent, gives the patient the opportunity to practice his responsibility and to decide the best for him in a friendly and confident environment. In the middle of science and technology, a close contact with ethics and humanism will allow a better understanding of the integrity of subjects, specially when they are ill. PMID- 15824840 TI - [Fever: conflicting scales]. PMID- 15824841 TI - [Heat-related illness: an underdiagnosed cause of relapsing fever ]. PMID- 15824843 TI - Revival of bone strength: the bottom line. PMID- 15824844 TI - Low skeletal muscle mass is associated with poor structural parameters of bone and impaired balance in elderly men--the MINOS study. AB - In 796 men, 50-85 years of age, decreased relative skeletal muscle mass index was associated with narrower bones, thinner cortices, and a consequent decreased bending strength (lower section modulus), as well as with impaired balance and an increased risk of falls. INTRODUCTION: In men, appendicular skeletal muscle mass (ASM) is correlated positively with BMC and areal BMD (aBMD). In elderly men, low muscle mass and strength (sarcopenia) is associated with difficulties in daily living activities. The aim of this study was to evaluate if ASM is correlated with bone size, mechanical properties of bones, balance, and risk of falls in elderly men. MATERIALS AND METHODS: This study used 796 men, 50-85 years of age, belonging to the MINOS cohort. Lifestyle factors were evaluated by standardized questionnaires. Estimates of mechanical bone properties were derived from aBMD measured by DXA. ASM was estimated by DXA. The relative skeletal muscle mass index (RASM) was calculated as ASM/(body height)(2.3). RESULTS: After adjustment for age, body size, tobacco smoking, professional physical activity, and 17beta estradiol concentration, RASM was correlated positively with BMC, aBMD, external diameter, and cortical thickness (r = 0.17-0.34, p < 0.0001) but not with volumetric BMD. Consequently, RASM was correlated with section modulus (r = 0.29 0.39, p < 0.0001). Men in the lowest quartile of RASM had section modulus of femoral neck and distal radius lower by 12-18% in comparison with men in the highest quartile of RASM. In contrast, bone width was not correlated with fat mass, reflecting the load of body weight (except for L(3)), which suggests that the muscular strain may exert a direct stimulatory effect on periosteal apposition. After adjustment for confounding variables, a decrease in RASM was associated with increased risk of falls and of inability to accomplish clinical tests of muscle strength, static balance, and dynamic balance (odds ratio per 1 SD decrease in RASM, 1.31-2.23; p < 0.05-0.001). CONCLUSIONS: In elderly men, decreased RASM is associated with narrower bones and thinner cortices, which results in a lower bending strength. Low RASM is associated with impaired balance and with an increased risk of falls in elderly men. It remains to be studied whether low RASM is associated with decreased periosteal apposition and with increased fracture risk in elderly men, and whether the difference in skeletal muscle mass between men and women contributes to the between-sex difference in fracture incidence. PMID- 15824845 TI - Relationship of volumetric BMD and structural parameters at different skeletal sites to sex steroid levels in men. AB - In a population-based, cross-sectional study, we related age-associated changes in vBMD and in bone structural parameters to circulating bioavailable estradiol and testosterone levels in men. Associations between these bone mass/structural parameters and sex steroid levels were progressively stronger with age. Our previously postulated "threshold" for skeletal estrogen deficiency was most evident at cortical sites. INTRODUCTION: Serum sex steroids, particularly estrogen levels, are associated with bone mass in men, and previous work has suggested that there may be a "threshold" bioavailable estradiol (bio E(2)) level below which the male skeleton becomes estrogen deficient. However, previous studies addressing this issue have exclusively used DXA, which cannot separate trabecular from cortical bone or provide information on bone geometry or structure. MATERIALS AND METHODS: In an age-stratified population sample of 314 men (age, 22-91 years), we assessed volumetric BMD (vBMD) and bone geometry by QCT at the lumbar spine, femoral neck, distal radius, and distal tibia and related these to circulating bio E(2) and bio testosterone (T) levels. RESULTS: Compared with young men (age, 20-39 years), middle-aged men (age, 40-59 years) had significantly lower bio T (-26%, p < 0.001) and bio E(2) (-9%, p = 0.038) levels, and these decreases were even greater in the elderly men (age > or = 60 years, -60% and -38% for bio T and bio E(2), respectively, p < 0.001 for both). Reflecting their intact gonadal status, vBMD/structural parameters were not related to sex steroid levels in young men, whereas bio E(2) levels were associated consistently with vBMD and variably with bone geometric parameters in the elderly men; middle-aged men showed associations with bio E(2) and bio T at some sites. At all cortical sites, vBMD was associated with bio E(2) at low (<30 pM, R = 0.27-0.41, p < 0.05-0.001) but not high (> or =30 pM, R = -0.003 to 0.12, p = not significant) levels; no such differences were evident at trabecular sites. CONCLUSIONS: In men, bio E(2) is the most consistent predictor of vBMD and some bone geometric variables as assessed by QCT. We also extend our previous findings on a possible "threshold" for skeletal estrogen deficiency by showing that this is most evident for cortical sites. PMID- 15824846 TI - Postmenopausal bilateral oophorectomy is not associated with increased fracture risk in older women. AB - We studied whether oophorectomy performed after menopause is associated with an increased risk of hip or vertebral fractures in 6295 Study of Osteoporotic Fractures participants. There was no association between postmenopausal oophorectomy and the risk of hip or vertebral fractures. INTRODUCTION: Bilateral oophorectomy after natural menopause has been associated with an increased risk of osteoporotic fractures, potentially because of a decline in serum estradiol and testosterone levels after the oophorectomy. We prospectively tested this hypothesis in the Study of Osteoporotic Fractures (SOF). MATERIALS AND METHODS: We studied 6295 white women 65 years of age participating in the SOF who were not taking estrogen therapy at baseline. Hip fracture analyses included 708 hip fractures; vertebral fracture analyses included 267 incident vertebral fractures. Baseline serum estradiol and free testosterone values were available in a small subset of participants. RESULTS AND CONCLUSION: There were no significant differences in age, weight, or BMD between the women who underwent postmenopausal oophorectomy (n = 583) and those who did not (n = 5712). Free testosterone levels were significantly lower among women who had a postmenopausal oophorectomy. A history of postmenopausal oophorectomy was not associated with an increased risk of hip (hazard ratio [HR] = 1.1; 95% CI = 0.9-1.5) or vertebral fracture (HR = 0.7; 95% CI = 0.5-1.2). The relationship between oophorectomy and hip fracture was not altered by adding serum estradiol level (HR = 1.3; 95% CI = 0.5-3.2) or serum free testosterone level (HR = 1.7; 95% CI = 0.8-3.7) to the model. In summary, postmenopausal oophorectomy was not associated with an increased risk of hip or vertebral fracture in this cohort. These results are in contrast to previous findings, suggesting that the relationship between postmenopausal oophorectomy and fractures is not fully elucidated and that incidental oophorectomy after menopause should still be considered carefully in each potential patient. PMID- 15824847 TI - Adjusting data to body size: a comparison of methods as applied to quantitative trait loci analysis of musculoskeletal phenotypes. AB - The aim of this study was to compare three methods of adjusting skeletal data for body size and examine their use in QTL analyses. It was found that dividing skeletal phenotypes by body mass index induced erroneous QTL results. The preferred method of body size adjustment was multiple regression. INTRODUCTION: Many skeletal studies have reported strong correlations between phenotypes for muscle, bone, and body size, and these correlations add to the difficulty in identifying genetic influence on skeletal traits that are not mediated through overall body size. Quantitative trait loci (QTL) identified for skeletal phenotypes often map to the same chromosome regions as QTLs for body size. The actions of a QTL identified as influencing BMD could therefore be mediated through the generalized actions of growth on body size or muscle mass. MATERIALS AND METHODS: Three methods of adjusting skeletal phenotypes to body size were performed on morphologic, structural, and compositional measurements of the femur and tibia in 200-day-old C57BL/6J x DBA/2 (BXD) second generation (F(2)) mice (n = 400). A common method of removing the size effect has been through the use of ratios. This technique and two alternative techniques using simple and multiple regression were performed on muscle and skeletal data before QTL analyses, and the differences in QTL results were examined. RESULTS AND CONCLUSIONS: The use of ratios to remove the size effect was shown to increase the size effect by inducing spurious correlations, thereby leading to inaccurate QTL results. Adjustments for body size using multiple regression eliminated these problems. Multiple regression should be used to remove the variance of co-factors related to skeletal phenotypes to allow for the study of genetic influence independent of correlated phenotypes. However, to better understand the genetic influence, adjusted and unadjusted skeletal QTL results should be compared. Additional insight can be gained by observing the difference in LOD score between the adjusted and nonadjusted phenotypes. Identifying QTLs that exert their effects on skeletal phenotypes through body size-related pathways as well as those having a more direct and independent influence on bone are equally important in deciphering the complex physiologic pathways responsible for the maintenance of bone health. PMID- 15824848 TI - Intravenous neridronate in children with osteogenesis imperfecta: a randomized controlled study. AB - In a randomized controlled study, we investigated the effect of treatment with intravenous neridronate in prepubertal children with OI. Our study suggests that quarterly intravenous infusions of the bisphosphonate significantly raise the rate of increase in BMD at both the spine and hip, the projected area of the lumbar vertebrae, and height. These results are associated with a significant decrease in the risk of clinical fractures. INTRODUCTION: Osteogenesis imperfecta (OI) is a heritable disease of connective tissue, characterized by increased bone fragility. Bisphosphonates currently seem to be the most promising therapy, but randomized, controlled studies are scarce and have never been carried out in prepubertal children. MATERIALS AND METHODS: This was a randomized, controlled 3 year clinical trial. The Italian Patients' Society of OI (AsItOI) sent their members affected by any type of OI to two centers at the University of Verona (Italy) to participate in the study. Sixty-four children, 6-11 years of age for boys and 6-9 years of age for girls, with no signs of puberty and who were never treated with bisphosphonates, were randomized to either intravenous neridronate (2 mg/kg infused IV in 30 minutes every 3 months) or no treatment, with a ratio of 2:1. Control patients were given the same bisphosphonate therapy at the end of the first year. BMD and projected bone areas, as measured by DXA, at spine and hip, height, and peripheral fracture incidence, both prospective and retrospective (2 years preceding randomization), were the main outcomes of the study. RESULTS: At the end of the first year, spine and hip BMD rose by 3.5-5.7% in control patients and by 18-25% (p < 0.001 versus controls) in the active group, respectively. During the following 2 years, the treatment in all patients was associated with BMD increases of 10-25% per year. Height and the DXA-derived projected area of lumbar spine rose during the first year of observation significantly more in the active group than in the control group (<0.01 and <0.05, respectively). Both height and spine projected area continued to rise in the treated patients toward levels found in healthy individuals. During the first year of treatment, 45% of the control patients and 27% of the active group had a nonvertebral fracture, but this difference was not statistically significant (p = 0.2). The total number of fractures was 18 in the 22 control patients and 13 in the active group (relative risk, 0.36; 95% CI, 0.15-0.87; p < 0.05). CONCLUSION: Intravenous neridronate infusions, administered quarterly, significantly increase BMD and lower the risk of clinical fracture in prepubertal children with OI. PMID- 15824849 TI - Decrease in thoracic vertebral bone attenuation with calcium-based phosphate binders in hemodialysis. AB - We performed a post hoc analysis of a 52-week randomized trial conducted in adult hemodialysis patients that compared the effects of calcium-based phosphate binders and sevelamer, a nonabsorbable polymer, on parameters of mineral metabolism and vascular calcification by electron beam tomography. In this analysis, we evaluated the relative effects of calcium and sevelamer on thoracic vertebral attenuation by CT and markers of bone turnover. Subjects randomized to calcium salts experienced a significant reduction in trabecular bone attenuation and a trend toward reduction in cortical bone attenuation, in association with higher concentrations of serum calcium, lower concentrations of PTH, and reduced total and bone-specific alkaline phosphatase. INTRODUCTION: In patients with chronic kidney disease, hyperphosphatemia is associated with osteodystrophy, vascular and soft tissue calcification, and mortality. Calcium-based phosphate binders are commonly prescribed to reduce intestinal phosphate absorption and to attenuate secondary hyperparathyroidism. Clinicians and investigators have presumed that, in hemodialysis patients, calcium exerts beneficial effects on bone. MATERIALS AND METHODS: We performed a post hoc analysis of a 52-week randomized trial conducted in adult hemodialysis patients that compared the effects of calcium-based phosphate binders and sevelamer, a nonabsorbable polymer, on parameters of mineral metabolism and vascular calcification by electron beam tomography. In this analysis, we evaluated the relative effects of calcium and sevelamer on thoracic vertebral attenuation by CT and markers of bone turnover. RESULTS AND CONCLUSIONS: The average serum phosphorus and calcium x phosphorus products were similar for both groups, although the average serum calcium concentration was significantly higher in the calcium-treated group. Compared with sevelamer-treated subjects, calcium-treated subjects showed a decrease in thoracic vertebral trabecular bone attenuation (p = 0.01) and a trend toward decreased cortical bone attenuation. More than 30% of calcium-treated subjects experienced a 10% or more decrease in trabecular and cortical bone attenuation. On study, sevelamer-treated subjects had higher concentrations of total and bone-specific alkaline phosphatase, osteocalcin, and PTH (p < 0.001). When used to correct hyperphosphatemia, calcium salts lead to a reduction in thoracic trabecular and cortical bone attenuation. Calcium salts may paradoxically decrease BMD in hemodialysis patients. PMID- 15824850 TI - Functional analysis of the single nucleotide polymorphism (787T>C) in the tissue nonspecific alkaline phosphatase gene associated with BMD. AB - Polymorphisms of the TNSALP gene have not previously been studied as a possible determinant for variations in BMD or as a predisposing genetic factor for osteoporosis. This study showed a significantly higher association between the 787T>C (Tyr246His) TNSALP gene and BMD among 501 postmenopausal women. Furthermore, the effects of amino acid substitution on the catalytic property of the protein translated from the 787T>C gene were examined. INTRODUCTION: Alkaline phosphatase (ALP) is present mainly on the cell membrane in various tissues and hydrolyzes a variety of monophosphate esters into inorganic phosphoric acid and alcohol. Human ALPs are classified into four types: tissue-nonspecific, intestinal, placental, and germ cell types. Based on studies of hypophosphatasia, which is a systemic skeletal disorder resulting from a tissue-nonspecific ALP (TNSALP) deficiency, TNSALP was suggested to be indispensable for bone mineralization. MATERIALS AND METHODS: We explored the possibility that the TNSALP gene may contribute to age-related bone loss in humans by examining the association between TNSALP gene polymorphisms and BMD in 501 Japanese postmenopausal women. To analyze the protein translated from the TNSALP gene associated with BMD, we constructed a TNSALP cDNA expression plasmid. RESULTS: We genotyped two single nucleotide polymorphisms (787T>C[Tyr246His] and 876A>G[Pro275Pro]), which proved to be in complete linkage disequilibrium. There was a significant difference in BMD and the BMD score adjusted for age and body weight (Z score) among haplotypes (p = 0.041), which was lowest among 787T/876A homozygotes, highest among 787T>C/876A>G homozygotes, and intermediate among heterozygotes. In subgroups divided by age, haplotypes were significantly associated with BMD in older postmenopausal women (>74 years; p = 0.001), but not in younger postmenopausal women (<74 years; p = 0.964). Expression of the 787T>C TNSALP gene using COS-1 cells showed that the protein translated from 787T>C had ALP-specific activity similar to that of 787T. Interestingly, the K(m) value for TNSALP in cells transfected with the 787T>C TNSALP gene was decreased significantly compared with that of cells bearing the 787T gene, reflecting the higher affinity. CONCLUSIONS: These results suggest that variation in TNSALP may be an important determinant of age-related bone loss in humans and that the phosphate metabolism pathway may provide a novel target for the prevention and treatment of osteoporosis. PMID- 15824851 TI - Heterozygous mutations in the LDL receptor-related protein 5 (LRP5) gene are associated with primary osteoporosis in children. AB - Three of 20 patients with juvenile osteoporosis were found to have a heterozygous mutation in the LRP5 gene. No mutations were found in the type I collagen genes. Mutations in the other family members with similar bone phenotype confirmed that LRP5 has a role in both juvenile and adult osteoporosis. INTRODUCTION: The gene encoding the low-density lipoprotein receptor-related protein 5 (LRP5) gene has recently been shown to affect bone mass accrual during growth and to be involved in osteoporosis-pseudoglioma syndrome and a high bone mass phenotype. Mutations in the type I collagen genes (COL1A1 and COL1A2) are known to cause osteogenesis imperfecta, characterized by increased bone fragility. MATERIALS AND METHODS: Here we analyzed COL1A1, COL1A2, and LRP5 for mutations in 20 pediatric patients with primary osteoporosis characterized by low BMD, recurrent fractures, and absent extraskeletal manifestations. RESULTS AND CONCLUSIONS: No mutations were detected in the type I collagen genes, but two missense mutations (A29T and R1036Q) and one frameshift mutation (C913fs) were found in the LRP5 gene in three of the patients. The frameshift mutation was also seen in the proband's father and brother, who both were found to have significant osteoporosis. R1036Q was observed in the proband's mother and two brothers, who all had osteoporosis. These results indicate that heterozygous mutations in the LRP5 gene can cause osteoporosis in both children and adults. PMID- 15824852 TI - Ghrelin directly regulates bone formation. AB - To clarify the role of ghrelin in bone metabolism, we examined the effect of ghrelin in vitro and in vivo. Ghrelin and its receptor, GHS-R1a, were identified in osteoblasts, and ghrelin promoted both proliferation and differentiation. Furthermore, ghrelin increased BMD in rats. Our results show that ghrelin directly affects bone formation. INTRODUCTION: Ghrelin is a gut peptide involved in growth hormone (GH) secretion and energy homeostasis. Recently, it has been reported that the adipocyte-derived hormone leptin, which also regulates energy homeostasis and opposes ghrelin's actions in energy homeostasis, plays a significant role in bone metabolism. This evidence implies that ghrelin may modulate bone metabolism; however, it has not been clarified. To study the role of ghrelin in skeletal integrity, we examined its effects on bone metabolism both in vitro and in vivo. MATERIALS AND METHODS: We measured the expression of ghrelin and growth hormone secretagogue receptor 1a (GHS-R1a) in rat osteoblasts using RT-PCR and immunohistochemistry (IHC). The effect of ghrelin on primary osteoblast-like cell proliferation was examined by recording changes in cell number and the level of DNA synthesis. Osteoblast differentiation markers (Runx2, collagen alpha1 type I [COLI], alkaline phosphatase [ALP], osteocalcin [OCN]) were analyzed using quantitative RT-PCR. We also examined calcium accumulation and ALP activity in osteoblast-like cells induced by ghrelin. Finally, to address the in vivo effects of ghrelin on bone metabolism, we examined the BMD of Sprague Dawley (SD) rats and genetically GH-deficient, spontaneous dwarf rats (SDR). RESULTS: Ghrelin and GHS-R1a were identified in osteoblast-like cells. Ghrelin significantly increased osteoblast-like cell numbers and DNA synthesis in a dose dependent manner. The proliferative effects of ghrelin were suppressed by [D Lys(3)]-GHRP-6, an antagonist of GHS-R1a, in a dose-dependent manner. Furthermore, ghrelin increased the expression of osteoblast differentiation markers, ALP activity, and calcium accumulation in the matrix. Finally, ghrelin definitely increased BMD of both SD rats and SDRs. CONCLUSIONS: These observations show that ghrelin directly stimulates bone formation. PMID- 15824853 TI - Inactivation of the Na-Cl co-transporter (NCC) gene is associated with high BMD through both renal and bone mechanisms: analysis of patients with Gitelman syndrome and Ncc null mice. AB - Chronic thiazide treatment is associated with high BMD. We report that patients and mice with null mutations in the thiazide-sensitive NaCl cotransporter (NCC) have higher renal tubular Ca reabsorption, higher BMD, and lower bone remodeling than controls, as well as abnormalities in Ca metabolism, mainly caused by Mg depletion. INTRODUCTION: Chronic thiazide treatment decreases urinary Ca excretion (UVCa) and increases BMD. To understand the underlying mechanisms, Ca and bone metabolism were studied in two models of genetic inactivation of the thiazide-sensitive NaCl cotransporter (NCC): patients with Gitelman syndrome (GS) and Ncc knockout (Ncc(-/-)) mice. MATERIALS AND METHODS: Ca metabolism was analyzed in GS patients and Ncc(-/-) mice under conditions of low dietary Ca. BMD was measured by DXA in patients and mice, and bone histomorphometry was analyzed in mice. RESULTS: GS patients had low plasma Mg. They exhibited reduced UVCa, but similar serum Ca and GFR as control subjects, suggesting increased renal Ca reabsorption. Blood PTH was lower despite lower serum ionized Ca, and Mg repletion almost corrected both relative hypoparathyroidism and low UVCa. BMD was significantly increased in GS patients at both lumbar (+7%) and femoral (+16%) sites, and osteocalcin was reduced. In Ncc(-/-) mice, serum Ca and GFR were unchanged, but UVCa was reduced and PTH was elevated; Mg repletion largely corrected both abnormalities. Trabecular and cortical BMD were higher than in Ncc(+/+) mice (+4% and +5%, respectively), and despite elevated PTH, were associated with higher cortical thickness and lower endosteal osteoclastic surface. CONCLUSIONS: Higher BMD is observed in GS patients and Ncc(-/-) mice. Relative hypoparathyroidism (human) and bone resistance to PTH (mice), mainly caused by Mg depletion, can explain the low bone remodeling and normal/low serum Ca despite increased renal Ca reabsorption. PMID- 15824854 TI - Bone adaptation to a mechanical loading program significantly increases skeletal fatigue resistance. AB - Using a mechanical loading program to induce bone adaptation, we found that small (<2-fold) changes in the structural properties of the rat ulna increased its fatigue resistance >100-fold. This indicates that a moderate exercise program may be an effective preventative strategy for stress fractures. INTRODUCTION: There are currently limited preventative strategies for stress fractures. Because stress fracture risk is directly influenced by skeletal properties, it has been hypothesized that modification of these properties using a mechanical loading program may positively influence risk. The aim of this study was to investigate whether the bone changes associated with a mechanical loading program can enhance skeletal fatigue resistance. MATERIALS AND METHODS: Site-specific mechanical loading was performed on one forearm of adult female Sprague-Dawley rats using the axial compression loading model. Loading was performed 3 days/week for 5 consecutive weeks to induce adaptation. The loaded and nonloaded ulnas in each animal were removed after the loading program, and their material and structural properties were determined. The ulna pairs were subsequently loaded until fatigue failure at the same constant peak axial load. RESULTS: Mechanical loading induced consistent and predictable changes in the structural properties of loaded ulnas, with the largest change being a nearly 2-fold increase in midshaft minimum second moment of area (I(MIN)). The mechanical-loading induced bone changes resulted in a >100-fold increase in fatigue resistance in loaded ulnas, with resistance being exponentially related to the structural properties of the ulna. CONCLUSIONS: This study found that by enhancing the structural properties of a bone through a mechanical loading program, its fatigue resistance could be significantly improved. This indicates that an exercise program aimed at modifying bone structure may be used as a possible prevention strategy for stress fractures. PMID- 15824855 TI - Genetic variation in structure-function relationships for the inbred mouse lumbar vertebral body. AB - Structure-function relationships were determined for L(5) vertebral bodies from three inbred mouse strains. Genetic variability in whole bone mechanical properties could be explained by a combination of the traits specifying the amount, distribution, and quality of the cortical and trabecular bone tissue. INTRODUCTION: Although phenotypically correlated with fracture, BMD may be disadvantageous to use in genetic and biomechanical analyses because BMD does not distinguish the contributions of the underlying morphological and compositional bone traits. Developing functional relationships between the underlying bone traits and whole bone mechanical properties should further our understanding of the genetics of bone fragility. MATERIALS AND METHODS: Microarchitecture and composition of L(5) vertebral bodies (n = 10/strain) from A/J, C57BL/6J, and C3H/HeJ inbred mouse strains were determined using muCT with an isotropic voxel size of 16 mum(3). Failure load, stiffness, and total deformation as a measure of ductility were measured in compression using a noncontact strain extensometer imaging system. A correlation analysis related morphological and compositional bone traits to whole bone mechanical properties. A multivariate analysis identified structure-function relationships for each genotype. RESULTS: No single bone trait accurately explained the genetic variation in mechanical properties. However, a combination of traits describing the amount, distribution, and quality of cortical and trabecular bone tissue explained >70% of the variation in vertebral mechanical properties. Importantly, structure-function relationships were unique among genotypes. CONCLUSIONS: Different genetic backgrounds use different combinations of underlying bone traits to create mechanically functional structures. Using a single complex trait such as BMD or BV/TV as the sole phenotypic marker in genetic analyses may prove to be disadvantageous because of the complex relationship between mechanical properties and the underlying bone traits. Therefore, considering multiple bone traits and the interaction among these bone traits is necessary to understand the relationship between genetic background and complex whole bone mechanical properties. PMID- 15824856 TI - Comparative effect of soy protein, soy isoflavones, and 17beta-estradiol on bone metabolism in adult ovariectomized rats. AB - This study provided a comprehensive investigation on the effect of soy protein and soy isoflavones on both calcium and bone metabolism in virgin adult rats. The measurements included bone histology, calcium kinetic modeling, calcium balance, bone densitometry, and whole body densitometry. Results confirmed the bone preserving effect of estrogen but did not support a bone-sparing role of soy isoflavones. INTRODUCTION: Several animal and short-term human studies have indicated that soy protein isolate enriched with isoflavones may be used as an alternative therapy to estrogen replacement therapy. However, none of the previous studies have investigated this estrogenic effect on both calcium and bone metabolism in animals or humans, which is essential in ascertaining the mode of action of isoflavones. MATERIALS AND METHODS: This study was designed to determine the effects of soy protein versus isoflavones on calcium and bone metabolism in an ovariectomized rat model. Unmated 6-month-old ovariectomized and sham-operated female Sprague-Dawley rats were randomly assigned to nine groups (16 rats/group) and pair-fed soy- or casein-based diets with or without isoflavones for 8 weeks. A reference group was administered estrogen through subcutaneous implants (20-35 pg/liter plasma). Bone densitometry, histomorphometry, and mechanical testing were used to study bone metabolism and quality. Calcium metabolism was studied using calcium tracer balance and kinetics. RESULTS: After ovariectomy, estrogen prevented bone loss in trabecular bone and suppressed formation on both trabecular and cortical bone surfaces. Isoflavones given as enriched soy protein isolate or supplements did not prevent trabecular bone loss. Combining isoflavones with estrogen had no additional benefits over estrogen alone. There were no differences in response to isoflavones caused by protein source. None of the treatments significantly affected either total Ca balance or (45)Ca absorption. However, soy protein showed significant effects on reducing urinary loss of Ca in animals, irrespective of isoflavone level, perhaps because of the lower amount of sulfur containing amino acids in soy protein. CONCLUSION: Estrogen, but not isoflavones at the levels tested, suppressed bone remodeling in both trabecular and cortical bone after ovariectomy. PMID- 15824857 TI - TRAF2 is essential for TNF-alpha-induced osteoclastogenesis. AB - TRAF2-deficient mice show embryonic lethality, and we developed a new in vitro differentiation system to show the function of TRAF2 in osteoclastogenesis, in which osteoclast progenitors are derived from the fetal liver of TRAF2-deficient mice. Using this system, we showed that TRAF2 is required for TNF-alpha-induced osteoclastogenesis. INTRODUCTION: TNF receptor-associated factor 2 (TRAF2) is a signal transducer for RANK and for two TNF receptor isotypes, TNFR1 and TNFR2. Because TRAF2-deficient mice show embryonic lethality, it has remained unclear whether TRAF2 is crucial in RANKL- or TNF-alpha-induced osteoclastogenesis. MATERIALS AND METHODS: Osteoclast progenitors derived from fetal liver were cultured in the presence of monocyte macrophage colony-stimulating factor (M CSF), and flow cytometry for characterization of surface markers on these cells was performed. To examine the involvement of TRAF2 in osteoclast differentiation, we cultured osteoclast progenitors from TRAF2-deficient and wildtype mice with soluble RANKL or TNF-alpha in the presence of M-CSF, and counted the number of TRACP(+) multinucleate cells formed. c-jun N-terminal kinase (JNK) and NF-kappaB activation in osteoclast progenitors was examined by Western blot analysis and electrophoretic mobility shift assay, respectively. Nuclear factor of activated T cells (NFATc1) expression and activation were analyzed by RT-PCR and immunofluorescence staining, respectively. To examine whether TRAF2 overexpression induced osteoclastogenesis, TRAF2 was overexpressed in osteoclast progenitors form wildtype bone marrow by retrovirus infection. RESULTS AND CONCLUSIONS: Osteoclast progenitors from normal fetal liver, which were cultured with M-CSF, expressed surface molecules c-fms, Mac-1, and RANK, and could differentiate into TRACP(+) multinucleate cells in the presence of soluble RANKL or TNF-alpha. RANKL-induced osteoclastogenesis gave a reduction of 20% in the progenitors from TRAF2-deficient mice compared with that of the cells from littermate wildtype mice, whereas TNF-alpha-induced osteoclastogenesis was severely impaired in the cells from the TRAF2-deficient mice. Only a few TRACP(+) multinucleate cells were formed, and TNF-alpha-mediated activation of JNK, NF kappaB, and NFATc1 was defective. TRAF2 overexpression induced differentiation of osteoclast progenitors from wildtype mice into TRACP(+) multinucleate cells. These results suggest that TRAF2 plays an important role in TNF-alpha-induced osteoclastogenesis. PMID- 15824858 TI - Combined angiogenic and osteogenic factor delivery enhances bone marrow stromal cell-driven bone regeneration. AB - Bone formation is a coordinated process involving various biological factors. We have developed a scaffold system capable of sustained and localized presentation of osteogenic (BMP-4) and angiogenic (VEGF) growth factors and human bone marrow stromal cells to promote bone formation at an ectopic site. Combined delivery of these factors significantly enhanced bone formation compared with other conditions. INTRODUCTION: Tissue regeneration entails complex interactions between multiple signals and materials platforms. Orchestrating the presentation of these signals may greatly enhance the regeneration of lost tissue mass. Bone formation, for example, is dependent on the signaling of BMPs, molecules initiating vascularization (e.g., vascular endothelial growth factor [VEGF]), and osteogenic precursor cells capable of responding to these cues and forming bone tissue. It was hypothesized that combined and concerted delivery of these factors from biodegradable scaffolds would lead to enhanced bone formation. MATERIALS AND METHODS: Poly(lactic-co-glycolic acid) scaffolds containing combinations of condensed plasmid DNA encoding for BMP-4, VEGF, and human bone marrow stromal cells (hBMSCs) were implanted into the subcutaneous tissue of SCID mice. Implants (n = 6) were retrieved at 3, 8, and 15 weeks after implantation. Bone and blood vessel formation was determined qualitatively and quantitatively by methods including histology, immmunostaining, and muCT. RESULTS: Scaffolds delivering VEGF resulted in a prominent increase in blood vessel formation relative to the conditions without VEGF. BMP-4 expression in scaffolds encapsulating condensed DNA was also confirmed at the 15-week time-point, showing the characteristic of long-term delivery in this system. Combined delivery of all three types of factors resulted in a significant increase in the quantity of regenerated bone compared with any factor alone or any two factors combined, as measured with DXA, X-ray, and histomorphometric analysis. Furthermore, bone formed with all three factors had elastic moduli significantly higher than any other condition. CONCLUSIONS: Concerted delivery of BMP-4, VEGF, and hBMSCs promoted greater bone formation relative to any single factor or combination of two factors. Materials systems that allows multifactorial presentation more closely mimic natural developmental processes, and these results may have important implications for bone regeneration therapeutics. PMID- 15824859 TI - Identification and characterization of cis-acting elements in the human and bovine PTH mRNA 3'-untranslated region. AB - The human PTH mRNA 3'-UTR has a cis element homologous to the rat cis-acting instability element and a more proximal element identical to the single binding element identified in bovine PTH mRNA 3'-UTR. The function of the elements was shown in vitro. INTRODUCTION: In the rat, Ca(2+) and phosphate regulate PTH mRNA stability by the interaction of trans-acting proteins with a defined cis-acting instability element in the distal region of the PTH mRNA 3'-untranslated region (UTR). This element has been characterized in the rat and is conserved in human, canine, feline, and murine 3'-UTRs but not in bovine and porcine 3'-UTRs. MATERIALS AND METHODS: Parathyroid protein-binding assays to the PTH mRNA transcripts were performed. Functionality was studied in reporter genes that were transiently transfected into HEK293 cells. RESULTS: Protein-RNA binding experiments identified an element in bovine PTH mRNA at the proximal end of the 3'-UTR that is different from the rat protein-binding element. The human 3'-UTR contains both elements, but only the distal element binds proteins. Functional studies with HEK293 cells transiently transfected with reporter genes containing the different elements and flanking nucleotides (nt) showed that the human distal element destabilized a reporter mRNA similar to the effect of this element in the rat. A reporter mRNA containing the single bovine PTH mRNA protein-binding element was also destabilized, and this was prevented by coexpression of AU-rich element binding factor 1 (AUF1). CONCLUSION: Our results identify a new protein binding element in the PTH mRNA 3'-UTR. In bovine PTH mRNA, it is the only element, and it is functional in destabilizing a reporter gene. It is also present in other species, including human PTH mRNA, where it is not functional, possibly because of differences in flanking sequences. The human PTH mRNA 3'-UTR distal element is highly homologous to the rat cis-acting instability element and destabilized a reporter gene, indicating its functionality. Therefore, different species have alternative cis-acting protein-binding elements that may determine the regulation of PTH mRNA stability in response to changes in serum calcium and phosphate. PMID- 15824860 TI - Retinoid signaling regulates CTGF expression in hypertrophic chondrocytes with differential involvement of MAP kinases. AB - Retinoids are important for growth plate chondrocyte maturation, but their downstream effectors remain unclear. Recently, CTGF (CCN2) was found to regulate chondrocyte function, particularly in the hypertrophic zone. The goal of the study was to determine whether CTGF is a retinoid signaling effector molecule, how it is regulated, and how it acts. INTRODUCTION: Using a combination of in vivo and in vitro approaches, we carried out a series of studies at the cellular, biochemical, and molecular level to determine whether and how retinoid signaling is related to expression and function of connective tissue growth factor (CTGF) in chondrocyte maturation and endochondral ossification. MATERIALS AND METHODS: Limbs of chick embryos in ovo were implanted with retinoic pan-antagonist RO 41 5253-filled beads, and phenotypic changes were assessed by in situ hybridization. CTGF gene expression and roles were tested in primary cultures of immature and hypertrophic chondrocytes. Cross-talk between retinoid signaling and other pathways was tested by determining endogenous levels of active ERK1/2 and p38 MAP kinases and phenotypic modulations exerted by specific antagonists of mitogen activated protein (MAP) kinases and BMP signaling (Noggin). RESULTS: Interference with retinoid signaling blocked expression of CTGF and other posthypertrophic markers in long bone anlagen in vivo and hypertrophic chondrocyte cultures, whereas all-trans-retinoic acid (RA) boosted CTGF expression and even induced it in immature proliferating cultures. Exogenous recombinant CTGF stimulated chondrocyte maturation, but failed to do so in presence of retinoid antagonists. Immunoblots showed that hypertrophic chondrocytes contained sizable levels of phosphorylated ERK1/2 and p38 MAP kinases that were dose- and time-dependently increased by RA treatment. Experimental ERK1/2 inhibition led to a severe drop in baseline and RA-stimulated CTGF expression, whereas p38 inhibition increased it markedly. These responses were gene-specific, because the opposite was seen with other hypertrophic chondrocyte genes such as collagen X and RA receptor gamma (RARgamma). Tests with Noggin showed that RA induction of CTGF expression was negatively influenced by BMP signaling, whereas induction of collagen X expression was BMP-dependent. CONCLUSIONS: Retinoids appear to have a preeminent role in controlling expression and function of CTGF in hypertrophic and posthypertrophic chondrocytes and do so with differential cooperation and intervention of MAP kinases and BMP signaling. PMID- 15824861 TI - Oropharyngeal skeletal disease accompanying high bone mass and novel LRP5 mutation. AB - Gain-of-function mutation in the gene encoding LRP5 causes high bone mass. A 59 year-old woman carrying a novel LRP5 missense mutation, Arg154Met, manifested skeletal disease affecting her oropharynx as well as dense bones, showing that exuberant LRP5 effects are not always benign. INTRODUCTION: Gain-of-function mutation (Gly171Val) of LDL receptor-related protein 5 (LRP5) was discovered in 2002 in two American kindreds with high bone mass and benign phenotypes. In 2003, however, skeletal disease was reported for individuals from the Americas and Europe carrying any of six novel LRP5 missense mutations affecting the same LRP5 protein domain. Furthermore, in 2004, we described a patient with neurologic complications from dense bones and extensive oropharyngeal exostoses caused by the Gly171Val defect. MATERIALS AND METHODS: A 59-year-old woman was referred for dense bones. Three years before, mandibular buccal and lingual exostoses (osseous "tori") were removed because of infections from food trapping between the teeth and exostoses. Maxillary buccal and palatal exostoses were asymptomatic. Radiographic skeletal survey showed marked thickening of the skull base and diaphyses of long bones (endosteal hyperostosis). BMD Z scores assessed by DXA were +8.5 and +8.7 in the total hip and L(1)-L(4) spine (both approximately 195% average control), respectively. LRP5 mutation analysis was carried out for the LRP5 domain known to cause high bone mass. RESULTS: Biochemical evaluation excluded most secondary causes of dense bones, and male-to-male transmission in her family indicated autosomal dominant inheritance. PCR amplification and sequencing of LRP5 exons 2-4 and adjacent splice sites revealed heterozygosity for a new LRP5 missense mutation, Arg154Met. CONCLUSIONS: LRP5 Arg154Met is a novel defect that changes the same first "beta-propeller" module as the eight previously reported LRP5 gain-of-function missense mutations. Arg154Met alters a region important for LRP5 antagonism by dickkopf (Dkk). Therefore, our patient's extensive oropharyngeal exostoses and endosteal hyperostosis likely reflect increased Wnt signaling and show that exuberant LRP5 effects are not always benign. PMID- 15824863 TI - Personality characteristics of asthma patients. AB - INTRODUCTION: Asthma, similarly to other chronic conditions, is strongly influenced by psychological factors. Previous studies have not established the personality characteristics of adult asthma patients as compared to non-patients. OBJECTIVES: To study the psychological characteristics of adult asthma patients in comparison to a pattern drawn up for the Portuguese population. To study the relationships between the personality and the severity and duration of the disease. METHODS: Outpatients of the Immunallergology and Pulmonology units of the Sao Joao Hospital suffering from asthma responded to the Revised NEO Personality Inventory (NEO-PI-R). Patients with co-morbidity factors were not excluded. The doctor who attended them classified the severity of the asthma according to the Global Initiative for Asthma (GINA). Relationships between the personality and the severity/duration of the disease were analysed using the ANOVA models. RESULTS: 300 asthma patients aged between 17 and 79 were studied. Of these, 75% were female; the great majority had intermittent/light persistent asthma (71%); 17% had moderate persistent asthma and 12% had severe persistent asthma. The duration of the disease was less than 10 years in 34% and over 23 years in 35%. The asthma patients had higher Neuroticism scores (p < 0.001), with the other facets (except impulsiveness) equally high. All the remaining domains- Extroversion, Openness to Experience, Conscientiousness and Agreeableness--had lower scores than the control group (p < 0.001). Both Extroversion and Openness to Experience decrease with growth in the severity (p = 0.003; p = 0.009) and the duration of the disease (p = 0.006; p = 0.013). Neuroticism increases in tandem with the severity of the disease. CONCLUSION: This study shows the predominance of Neurotic characteristics and lowered Extroversion, Openness to Experience, Agreeableness and Conscientiousness characteristics in asthmatics as compared to the general Portuguese population. Decreased Extroversion and Openness to Experience are observed as the severity and duration of the disease increase. Further studies are necessary to clarify the relationships between personality and the severity and duration of the disease. PMID- 15824864 TI - Acoustic rhinometry, rhinomanometry and spirometry of the asthmatics patients with rhinitis symptoms before and after oral corticoid. AB - INTRODUCTION: Asthma and rhinitis frequently co-exist in the same patient. It may exist a specific condition witch affects the general respiratory activity and also able to produce symptoms into upper and/ or lower airways. PURPOSE: To analyze the alteration and functional burst of upper and lower airways before and after oral corticoid. METHODS: From July of 2000 to August of 2002, sixteen (69 % was female ), acute asthmatics patients with rhinitis symptom assisted in the pneumology department, were analyzed through cross-sectional study using: symptoms questionnaire, physical examination, radiogram, spirometry, acoustic rhinometry, and rhinomanometry. RESULTS: Sixteen patients presented compromising symptoms of the upper airways. In the spirometry, FEV1, FEV1/FVC e FEF25-75%, presented major statistically differences. The acoustic rhinometry presented important statistical differences in the minimal cross- sectional area 1 e 2 in the left side and total nasal volume, after medicine intervention. CONCLUSIONS: The functional evaluations upon lower and upper airways, improvements in FEV1, FEV1/FVC e FEF25-75%, were observed through spirometry and total nasal volume, analyzing the acoustic rhinometry after oral corticoid. PMID- 15824865 TI - [Advances and challenger in the diagnosis and medical treatment of respiratory diseases]. AB - In the present article, we reviewed the most recent advances in the medical area and areas that contributed for the advances of medical science. We relate the advances in the areas of Biology and the Genetics, Bioengineering and Telecommunications among others. We also reviewed the most relevant advances in some areas of the more frequent pathologies of the respiratory tract, particularly those that urgently need new progress in terms of new techniques of diagnosis, therapeutics or prevention measures in the next years. PMID- 15824866 TI - [Influenza B virus pneumonia after bone marrow transplantation: case report with emphasis to the high resolution computed tomography findings]. AB - Nine year-old male patient, who underwent bone marrow transplantation because of acute myelogenous leukemia, presented with fever and dry cough three days after the procedure. The chest radiograph demonstrated bilateral diffuse reticular infiltrate. The high resolution computed tomography showed peribronchovascular interstitial thickening, peripheral small centrilobular nodules and areas of ground-glass attenuation. The bronchoalveolar lavage demonstrated positive direct fluorescence antibody testing against influenza B virus. Treatment with aerolizated ribavirin was instituted during 10 days and the patient showed clinical-radiological improvement. PMID- 15824867 TI - [Bronchospasm and hemoptysis. Could have the same cause?]. AB - The authors describe the case of a man, 46 years old, former smoker, former bird breeder, followed in the service of Imunoallergology for bronchospasm without response to the treatment. After experiencing an hemoptysis he was admitted in our service. He was submitted to many exams, which include a flexible bronchoscopy. This technique allows the identification of an endobronquial necrotic mass, which almost obstructed the main right bronchus. In order to obtain a bigger and more representative sample a rigid bronchoscopy was performed. This technique showed a broncholith, which was partially removed, and a bronchial fistula. Actually he is assimptomatic and under annual surveillance. First described for Aristoteles, the broncholithiasis is defined by the presence of stones in the bronchial tree. It is a very rare pathology, with a variable clinical expression, which must be considered whenever there is respiratory complaints in association with hilar or mediastinic calcifications. PMID- 15824868 TI - [Churg-Strauss syndrome--case report]. AB - Churg-Strauss syndrome (CSS) is a rare systemic vasculitis, that essentially affects medium-sized vessels, involves multiple organs and in the majority of cases is related to asthma and eosinophilia. The authors make a review of this disease and discuss its differential diagnosis presenting the case of a woman with asthma that at the age of 65 is diagnosed with CSS. PMID- 15824869 TI - [Cardiac resynchronization therapy: are all questions answered?]. PMID- 15824870 TI - [Acute hemodynamic effects]. AB - Several studies on the acute effect of cardiac resynchronization in patients with advanced heart failure (HF) and left bundle branch block (LBBB) have shown that left and biventricular stimulation increase pulse pressure and contractility, while patients with a QRS complex <150 ms may deteriorate during stimulation. Patients with LBBB, severe HF and a QRS width >150 ms underwent right, left and biventricular stimulation at different AV delays. Acute response was defined as > or =10% pulse pressure increase. 165 of 188 patients (88%) in sinus rhythm (47 women, mean age 62.5+/-10 years, ejection fraction 23+/-8%, NYHA class 3.1+/-0.3) were regarded acute responders. 10% of 103 patients with dilated cardiomyopathy and 16.5% of 79 patients with coronary artery disease were considered non responders. 29 patients (81%) with 2 posterolateral veins were acute responders with 10 of them (33%) being responders in only one vein. 54 patients had a higher pulse pressure increase (10.7+/-10.6%) with atrio-left ventricular stimulation, 48 patients with atrio-biventricular stimulation (9.8+/-6.4%). At one-year follow up, heart failure had significantly (p<0.0001) improved from NYHA class 3.1+/-0.4 to 2.1+/-0.7, VO(2)peak from 12.7+/-2.8 to 15.9+/-3.6 ml/min/kg. Left ventricular enddiastolic diameter being an indicator of reverse remodeling decreased from 80.5+/-10.5 to 73.3+/-13 (p<0.0001). Hemodynamic testing before CRT allows for the identification of acute non-responders as well as the best mode and site of stimulation and the optimal atrioventricular delay in responders. PMID- 15824871 TI - [Improved identification of suitable patients for cardiac resynchronization therapy by transthoracic echocardiography]. AB - Transthoracic echocardiography provides numerous options for the evaluation and quantification of contractile cardiac asynchrony in patients with advanced heart failure. Important information on the presence of asynchrony can be obtained already during a standard routine examination with conventional techniques (2D, M mode and Doppler). Newer techniques such as tissue Doppler imaging and real-time 3D-echocardiography enable us to better quantify the degree of asynchrony. The following article describes the echocardiographic features of asynchrony and algorithms for the improved identification of suitable patients for cardiac resynchronization therapy. PMID- 15824872 TI - [Effect of cardiac resynchronization therapy (CRT) on exercise tolerance, functional capacity and quality of life in patients with congestive heart failure]. AB - This review article summarizes the effect of cardiac resynchronization therapy (CRT) on exercise tolerance, functional capacity and quality of life, as it has been shown in previous randomized controlled trials. Based on these data we carefully have to reconsider the initial goals of this therapy. An early prophylactic implantation is not justified today. Especially patient selection has to be performed with more care in the future using new methods for detection of cardiac asynchrony to avoid a mismatch between implant numbers and real functional benefit. PMID- 15824873 TI - [Cardiac resynchronization therapy and arrhythmias]. AB - Cardiac resynchronization therapy (CRT) is now considered an established therapy for patients with chronic heart failure in the presence of a wide QRS complex. Though proarrhythmic effects have been described in a few cases, CRT did not increase the frequency of ventricular tachyarrhythmias in prospective studies. In patients on CRT therapy, persistent atrial fibrillation sometimes converts back to sinus rhythm, possibly dependent on the duration of atrial fibrillation. PMID- 15824874 TI - [Effect of CRT on morbidity and mortality]. AB - Patients with chronic heart failure who show a left ventricular ejection fraction < or =35% and remain in NYHA class III or IV despite optimal pharmacologic treatment show less morbidity and mortality on cardiac resynchronization therapy (CRT) if the left ventricle shows asynchrony. Although only one study has shown a significant reduction of mortality about 4% less deaths per year in the first two years can be resumed. Procedure related mortality is less than 1%. By improving on average one NYHA class 10-20% less patients experience hospitalization due to heart failure in the first two years after implantation of a CRT device. Patients who are hospitalized despite CRT have on average a hospital stay below 5 days. At least 10% of patients are currently suffering surgical revisions due to infections, dislocations and high pacing thresholds of the left ventricular lead. If costs saved by less hospitalization can finance costs of CRT or even exceed the costs of CRT remains unknown. PMID- 15824875 TI - [Current guidelines]. AB - Cardiac resynchronisation therapy (CRT) has been adopted fast into clinical practice and current guidelines as a new therapy for some patients with severe systolic heart failure. Current American guidelines have accepted cardiac resynchronisation therapy with the highest level of evidence (A) and have classified the indication in the second highest range (II a). Recommendations in a consensus statement of the German Cardiac Society have been more moderate and have considered contemporarily unsolved questions. After completion of the COMPANION study, which showed a significant benefit for cardiac-resynchronization therapy in the primary end point (mortality and hospitalisation), there were claims for a new evaluation of the cardiac resynchronisation therapy in the German guidelines for cardiac pacing. Contrarily to the American guidelines a risk stratification has been performed into class I (therapy should be performed), class IIa (therapy is reasonable) and class II b (therapy is not unreasonable). PMID- 15824877 TI - [Cardiac resynchronization therapy--what will the future bring?]. PMID- 15824876 TI - [Peculiarities in the follow up of resynchronization therapy]. AB - While resynchronization therapy (CRT) is based on conventional pacemaker- and ICD technology, its main intention is to minimize inter- and intraventricular asynergy and to establish optimal AV timing if sinus rhythm is preserved. The focus of this contribution is a series of conditions which jeopardize the therapeutic goal of CRT and should be recognized and hopefully corrected during follow up of CRT systems. These scenarios include uncertainties about left ventricular capture, double sensing in the ventricles, inhibition of the ventricular output (and loss of resynchronization), atrial and ventricular tachycardia and rate adaptation. Technical issues of following rhythm management devices are only discussed in the context of CRT requirements. PMID- 15824878 TI - [Pacemaker---EKG--Quiz]. PMID- 15824881 TI - Subfrontal schwannoma. AB - Subfrontal schwannoma is a rare disease, which can be mis-diagnosed as an olfactory meningioma or a neuroblastoma, because of similar clinical symptoms and signs and neuroradiological features. Especially for young subjects, olfactory neuroblastoma should be carefully differentiated, since the management strategies for those lesions are significantly different. The craniofacial approach is often needed for the resection of a neuroblastoma. We report a case of 14-year old boy in which olfactory neuroblastoma was suspected prior to surgery, but turned out to be a schwannoma histologically. Molecular genetic examination revealed neither NF2 gene mutation nor loss of heterozygosity of chromosome 22q, unlike common schwannomas. PMID- 15824880 TI - Review and critique of the quality of exercise recommendations for cancer patients and survivors. AB - The benefit of exercise for cancer patients is starting to become recognized. The purposes of this paper were to review the literature to examine whether research findings are being converted into guidelines for patients and survivors and to examine the quality of evidence on which they were based. A computer search of major health databases was conducted for peer-reviewed literature and books on exercise and cancer, and an Internet search was conducted for cancer websites reporting any exercise guidelines/recommendations for cancer patients. Seven peer reviewed articles, eight books and eight cancer websites were identified that suggested exercise guidelines for cancer patients and survivors. None of the published guidelines identified appeared to have been developed via a process that would allow them to be cited as evidence-based guidelines. Based on the studies to date, no direct cancer-specific evidence about the best type, frequency, duration or intensity of exercise is currently available in the peer reviewed literature. It is currently not known what would be most beneficial for which cancers, at which stage of disease or treatment. Given the current interest in cancer and exercise, there is an urgent need for an evidence-based set of exercise guidelines to be developed. PMID- 15824882 TI - Extracellular fluid S100B in the injured brain: a future surrogate marker of acute brain injury? AB - The authors describe the measurement of S100B protein in brain extracellular fluid (ECF) of patients with acute brain injury (traumatic brain injury and subarachnoid haemorrhage) using the technique of microdialysis. To our knowledge, this is the first report of S100B measurement in the human brain. Acute Brain Injury (ABI) is a leading cause of death and disability and the need for a practical and sensitive biochemical marker for monitoring these patients is urgent. The calcium binding astrocyte protein, S100B, may be a candidate for this role. Previous serum studies have shown S100B to be a sensitive predictor of mortality and rise in intracranial pressure in ABI, but it has never before been measured directly within the brain. The ECF reflects the local biochemistry of the brain parenchyma, and the use of intracerebral microdialysis opens up the possibility of studying many novel surrogate markers of injury in the laboratory, in addition to the conventional markers it measures at the bedside (lactate, pyruvate, glucose, and glycerol). In this preliminary report of two cases, the authors demonstrate the quantification of S100B in ECF microdialysate, and investigate whether changes in hourly S100B profile can be related to secondary brain injury. It is shown that extracellular concentrations of S100B change markedly in response to secondary brain injury. Further investigation is required to determine whether extracellular S100B measurement in ABI could assist in patient management. PMID- 15824883 TI - Effects of deletions of kelch-like genes on cowpox virus biological properties. AB - Cowpox virus (CPXV) strain GRI-90 contains six genes encoding kelch-like proteins. All six proteins contain both, the N-terminal BTB domain and the C terminal kelch domain. We constructed mutant variants of a CPXV strain with targeted deletions of one to four genes of the kelch family, namely D11L, C18L, G3L, and A57R. As kelch genes are located in terminal variable regions of the CPXV genome, we studied the relationship of these genes with integral biological characteristics such as virulence, host range, reproduction in vitro and in ovo (in chicken embryos). It was demonstrated that the following effects occurred in a gene dose dependent manner with an increase of the number of genes deleted: (1) range of sensitive cells altered--deletion mutants lacking three genes displayed a considerably decreased ability to reproduce in MDCK cells; mutants lacking four genes lost this ability completely; (2) analysis of pocks formed by mutants with deletion of three and four kelch-like genes on chorioallantoic membranes of chicken embryos demonstrated that pock size and virus yield were significantly decreased; (3) light microscopic analysis of the pocks revealed impaired proliferation and reduced vascularisation in the pock region. More alterations were detected by electron microscopic analysis: the reproduction of mutants results in a reduction of the number of mature virions formed, and in many cells this process was arrested at the stage of assembly of immature virions; and (4) the evaluation of LD(50) and body weight loss in BALB/c mice infected intranasally with CPXVs revealed a reduction of the virulence of the deletion mutants, which became statistically significant when four kelch-like genes were excised. PMID- 15824884 TI - Nucleocapsid gene variability reveals two subgroups of Lettuce necrotic yellows virus. AB - The complete nucleocapsid (N) genes of eight Australian isolates of Lettuce necrotic yellows virus (LNYV) were amplified by reverse transcription PCR, cloned and sequenced. Phylogenetic analyses of these sequences revealed two distinct subgroups of LNYV isolates. Nucleotide sequences within each subgroup were more than 96% identical but heterogeneity between groups was about 20% at the nucleotide sequence level. However, less than 4% heterogeneity was noted at the amino acid level, indicating mostly third nucleotide position changes and a strong conservation for N protein function. There was no obvious geographical or temporal separation of the subgroups in Australia. PMID- 15824885 TI - Stable deletions arising in the readthrough region of Soil-borne wheat mosaic virus RNA2 define the 5' limit of the functional promoter for the p19 subgenomic RNA. AB - The appearance of de novo deletion mutations in the readthrough (RT) region (nucleotide positions 861-2591) downstream of the capsid protein (CP) gene of a Japanese strain of Soil-borne wheat mosaic virus RNA2 was examined using infectious transcripts. Mutant RNA2s with different deletions predominated in independent serial passage experiments but all best-adapted mutants retained the 3'-terminal portion of the RT gene in frame with the CP gene. The longest best adapted mutation deleted the 1434 nucleotides between positions 1061 and 2494. When the RT protein was truncated by insertion of a termination codon plus an additional nucleotide to give a +1 frame-shift, after serial passages the progeny viruses regained the ability to express the C-terminal region of RT by an internal deletion. The 5' terminus of the p19 subgenomic RNA was identified at position 2598 and an essential transcription signal for this mRNA mapped between positions 2534 and 2563. A mutant in which this essential promoter element has been deleted cannot transcribe the p19 subgenomic RNA and has lost infectivity in planta. These results indicate that the 3'-terminal region of the RT gene has a major function in cis for expression of p19, which is essential for infecting plants. A reason for retaining the RT C-terminal region in stable deletion mutants is still unknown. PMID- 15824886 TI - Observations on recovery from and recurrence of HSV-2 infections in adult mice that were rescued from lethal vaginal infection by antiviral therapy. AB - An adult mouse model for studies of latency and recurrence after vaginal HSV-2 infection is not available at present, largely because the infection kills most mice within 14 days. We describe here an antiviral therapy that rescues most vaginally infected mice from death. Vaginally infected mice were nearly all rescued by combined treatment with one dose of monoclonal anti-HSV glycoprotein D 3 days after infection plus valacyclovir in the drinking water on days 3, 4, 5, 7, 9, 11, 13, and 15 after infection. At 60 days after infection, PCR measurements revealed that most rescued mice had viral DNA in their lumbosacral dorsal root ganglia, lumbosacral spinal cords, and paracervical autonomic ganglia, consistent with the possibility that latent infections were established. At this time, immunolabeling revealed CD45+ lymphoid cells in these neural tissues in rescued mice but not in normal control mice. In vivo depletion of T lymphocytes with monoclonal antibodies caused a recurrence of herpes illness symptoms earlier and in a larger proportion of rescued mice than was observed in non-depleted rescued mice. Interestingly, many rescued mice (46/114) spontaneously developed a syndrome of typical herpes illness symptoms that began with ruffled fur on a mouse that previously had sleek fur and progressed to arched backs, feeble gait, hindlimb paralysis, and death or euthanasia, or in some cases to recovery to health. This high incidence of apparent spontaneous recurrence of HSV-2 infection in rescued mice suggests that it may be possible, with some refinement of the procedure, to obtain an effective adult mouse model for studies of therapeutic vaccination to inhibit or prevent HSV-2 recurrence after genital tract infection. PMID- 15824887 TI - Biolistic infection of cassava using cloned components of Indian cassava mosaic virus. AB - Cassava mosaic disease (CMD) is a major constraint to cassava production in Africa and Asia. Of the two begomoviruses associated with CMD on the Indian subcontinent, Indian cassava mosaic virus (ICMV) and Sri Lankan cassava mosaic virus, only the latter has been successfully reintroduced into cassava to resolve the aetiology of the disease. Here, we report the complete nucleotide sequence of an ICMV isolate from Maharashtra (ICMV-[Mah2]), central India. Biolistic inoculation of the cloned components produced a systemic infection and typical mosaic symptoms in cassava, thereby fulfilling Koch's postulates. The availability of infectious clones will provide a valuable tool to screen new cassava cultivars for disease resistance under defined conditions. PMID- 15824888 TI - Expression, antigenicity and studies on cell receptor binding of the hemagglutinin of infectious salmon anemia virus. AB - Infectious salmon anemia (ISA) virus belongs to the proposed genus Isavirus of Orthomyxoviridae and is an emerging pathogen in Atlantic salmon (Salmo salar) farming. The hemagglutinin-esterase (HE) of ISA virus share several characteristics with the influenza virus hemagglutinin. This study reports recombinant expression of different ISA virus HE mutants in fish cell lines. Some introduced mutations, representing minimal differences in single amino acid residues, resulted in remarkable effects on expression efficiency in general and on surface expression specifically. Receptor binding assays showed that amino acid residues in the N-terminal half part are important in receptor binding, either being directly involved in the binding, or by influencing the structure of the binding site. Deletion of the putative N-glycosylation sites of the ISA virus HE, located near the transmembrane region, did not influence expression, receptor binding properties or staining by either a neutralising MAb, or salmon convalescent sera. The humoral immune response of Atlantic salmon after ISA virus infection showed weak neutralising activity and the results indicated that it was directed against HE. PMID- 15824889 TI - Increased bone density and decreased bone turnover, but no evident alteration of fracture susceptibility in elderly women with diabetes mellitus. AB - Bone density, bone turnover and fracture susceptibility were evaluated in 1,132 randomly recruited women, all 75 years old. Seventy-four of the women had diabetes, while 1,058 women did not. Areal bone mineral density (aBMD) of the hip and lumbar spine was investigated by dual energy X-ray absorptiometry (DXA), and bone mass of the calcaneus was measured by ultrasound. Urinary deoxypyridinoline/creatinine (U-DPD/Crea) and serum C-terminal cross-linked telopeptide of type 1 collagen (S-CTX) were assessed as markers of bone resorption. Serum bone-specific alkaline phosphatase (S-bone ALP) and serum osteocalcin (S-OC) were assessed as markers of bone formation. Also, serum 25(OH) vitamin D and serum parathyroid hormone (S-PTH) were assessed. Fracture susceptibility was evaluated retrospectively and prospectively for up to 6.5 years. In diabetic women, the aBMD of the femoral neck was 11% higher (p<0.001), and BMD of the lumbar spine was 8% higher (p=0.002) than in non-diabetic women. There was no difference in bone mass by ultrasound of the calcaneus. Women with diabetes had higher BMD of the femoral neck (p<0.001) and lumbar spine (p=0.03) also after correction for differences in body weight. In diabetic women, U DPD/Crea, S-CTX, and S-OC were decreased when compared with non-diabetic women (p=0.001 or less). After correction for covariance of body weight and plasma creatinine, S-CTX (p<0.001) and S-OC (p<0.001) were still lower in the diabetic women. Diabetic patients had hypovitaminosis D (p=0.008), a difference explained by differences in time spent outdoors and body weight. S-PTH did not differ between the groups. Women with diabetes had no more lifetime fractures (52%) than women without diabetic disease (57%), (p=0.31). This study shows that elderly women with diabetes and without severe renal insufficiency have high bone mass and low bone turnover. The high bone mass and low bone turnover is not likely to have a strong influence on fracture susceptibility. PMID- 15824890 TI - Osteoporosis In Turkish HIV/AIDS patients: comparative analysis by dual energy X ray absorptiometry and digital X-ray radiogrammetry. AB - Recently, osteoporosis has attracted concern among physicians treating patients with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS). Many confounding factors are assumed to play a role in its pathogenesis. The discussion has increased dramatically since the introduction of highly active antiretroviral therapy (HAART), and, in fact, the pertinent data have become much more contradictory. In this study, we have evaluated the BMD of our HIV/AIDS patients, comparatively by two methodologies: dual energy X-ray absorptiometry (DXA) and digital X-ray radiogrammetry (DXR). The study comprised 27 HIV/AIDS patients (15 males, 12 females). Bone mineral density measurements using DXA (Hologic QDR-4500) were performed at the lumbar spine (L1-L4), femur and distal radius. DXR evaluations were done by Pronosco X-posure system (Sectra Pronosco, Denmark) using the X-ray graphs of the patients' non-dominant hands. Nine patients (33.33%) were found to have osteoporosis. Fourteen (51.85%) had osteopenia and four (14.81%) were normal. Estimated BMD and cortical thickness measurements obtained from DXR significantly correlated with lumbar, femoral and radial DXA measurements. Whether disease-related or drug-related, the tendency to a decline in the bone mass of these patients exists, and future studies are awaited to unravel the clinical significance of osteopenia, risk of fracture and the individual differences between HAART regimens in HIV/AIDS. We also believe that launching screening and treatment guidelines for osteoporosis in these patients will then become reasonable. Last but not least, DXR appears to be a promising tool in this regard. PMID- 15824891 TI - Identification of 156 novel SNPs in 29 genes encoding G-protein coupled receptors. AB - We have been performing extensive screening on single nucleotide polymorphisms (SNPs) in and around genes encoding drug metabolizing enzymes, transporters, and receptors and have constructed the high-density SNP maps of such gene regions. In addition to genetic information reported earlier, we identified a total of 390 genetic variations, 358 SNPs and 32 genetic variations of other types, detected in 29 genes encoding G-protein coupled receptors in Japanese populations. Following a comparison of our data with SNPs in the dbSNP database in the US National Center for Biotechnology Information, 156 SNPs from these gene loci are considered to be novel. The fine-scale SNP maps constructed in this study should serve an important resource for studies of linkage-disequilibrium mapping for complex genetic diseases and drug-response phenotypes. PMID- 15824892 TI - Abrogation of DUSP6 by hypermethylation in human pancreatic cancer. AB - Our previous study indicated that DUSP6/MKP-3/PYST1 could act as a tumor suppressor in human pancreatic cancer. DUSP6 was frequently underexpressed in primary pancreatic cancer tissues by an unknown mechanism. In this study, we demonstrated that hypermethylation of the expressional control region of DUSP6 could account for its abrogation in cultured human pancreatic cancer cells and in primary pancreatic cancer tissues. First, we checked intrinsic transcriptional expression levels of DUSP6 by a quantitative real time PCR assay in 16 cultured pancreatic cancer cell lines and found that the cells could be classified into four groups: very-low-level expression, low-level expression, high-level expression, and very-high-level expression. We observed restored expression of DUSP6 after treatment with 5-azacytidine and trichostatin A, a DNA methyltransferase inhibitor and a histone deacetylase inhibitor, respectively, in cells with intrinsically very-low-level and low-level expression of DUSP6. Using a sodium-bisulfite-modification assay, we found that CpG sequences in intron 1 of DUSP6 were heavily methylated in MIA PaCa-2 and PAN07JCK, both showing the very low level of intrinsic expression of the gene. On the other hand, no methylation in this region was detected in 14 other cell lines. We checked the methylation state of this region by a methylation-specific PCR method in 12 primary pancreatic cancer tissues and compared it with the expression state of DUSP6 investigated by immunohistochemistry. Methylation was detected in five of eight cases with abolished expressions of DUSP6, four of which were poorly differentiated adenocarcinoma. On the other hand, none of the four cases with preserved expression of DUSP6 showed methylation. The methylation state significantly correlated with both the abolishment of protein expression (p = 0.038) and the histological subtype of adenocarcinoma (p = 0.023) by chi-square test. These results indicate that hypermethylation of the CpG islands in intron 1 may account for the strong suppression of DUSP6 expression. Other mechanism(s) and/or other CpG sites outside of our investigation may have some influence upon expressional suppression. Our combined results suggest that hypermethylation with modification of histone deacetylation play an important role in transcriptional suppression of DUSP6 in human pancreatic cancer. PMID- 15824893 TI - Snf1 kinase complexes with different beta subunits display stress-dependent preferences for the three Snf1-activating kinases. AB - Three upstream kinases, Pak1, Tos3 and Elm1, are able to activate the Snf1 kinase. Since the Snf1 kinase itself assembles into three complexes that differ in their beta subunit identity, the possibility exists that each upstream kinase might be dedicated to a single isoform of the Snf1 kinase. To test this dedicated activator hypothesis, we generated a series of yeast strains that lacked different combinations of upstream kinases and beta subunits. Cells expressing only one of the three upstream kinases exhibited distinct abilities to activate Snf1, depending on the beta subunit present in the Snf1 kinase complex and the stress imposed on the cells. Pak1 and Gal83 were the most promiscuous. Pak1 was able to activate all three isoforms of the Snf1 kinase under all stress conditions tested. The Gal83 isoform of Snf1 was able to be activated by any of the three upstream kinases under aerobic growth conditions but showed a preference for Pak1 during growth on raffinose. Our results indicate that the three Snf1-activating kinases are not dedicated to specific isoforms of the Snf1 kinase. Instead, the different isoforms of the Snf1 kinase display stress dependent preferences for the Pak1, Tos3 and Elm1 kinases. PMID- 15824894 TI - [The citation frequency for prominent researchers in HNO medical science. Scientometric follow-up of German scientists over 10 years]. AB - BACKGROUND: In 1995, 18 scientists involved in research on otorhinolaryngology in Germany (median of age 55 years; one woman) entered the Who is Who of German medical research because of their top citation rates in the worldwide Science Citation Index (SCI). Their publications were cited about 12 times as often as those of colleagues with an habilitation or professorship. Are their publications equally important in international research activity 10 years later? METHODS: Investigations were carried out using SCI 2002. RESULTS: Seventeen of the former stars still publish from Germany. Their total citation rate has remained almost unchanged. CONCLUSIONS: The stability and the large individual differences in citation rates indicate that a personal dimension is present, here called "research-talent", i.e. the ability to produce research of high quality. Internationally highly successful researchers reach their maximum productivity at about 40 years of age and maintain it even after official retirement. Therefore, there is little risk in promoting them and their projects. Moreover, they serve as examples for their colleagues and the rising scientific generation on how to do internationally successful research. PMID- 15824895 TI - [Impact of rescue method and the destination clinic on mortality in polytrauma. A status report]. AB - BACKGROUND: The aim of this study was to document the present knowledge from the medical literature on (1) efficacy of aeromedical evacuation (helicopter emergency medical service, HEMS) and (2) influence of the level of the first receiving hospital on mortality of patients. METHODS: Systematic review of the literature between 1970 and 2003; identification of studies with an evidence level of at least III and included control group; own results. RESULTS: (1) 17 studies concerning the efficacy of HEMS were included into the review. No single study yielded shorter rescue times with the use of HEMS. 11 of 17 studies showed a significantly higher survival rate (8.2 to 52%) with the employment of HEMS especially with mid-degree polytrauma. (2) All 6 relevant studies dealing with hospital level found a considerable lower mortality rate (19 to 42%) for patients treated primarily at a level 1 trauma center or comparable institution. CONCLUSIONS: The analyzed studies showed a trend toward decreased mortality rates with the employment of HEMS. Considering the comparable hospital level and even longer rescue times with HEMS, these differences can be explained with higher quality of initial diagnosis and treatment of the HEMS rescue team. Furthermore, mortality rates can be lowered significantly through primary treatment at a level 1 trauma center. Thus, the more flexible choice of the first receiving hospital represents a specific, clinically relevant advantage of HEMS in emergency medicine. PMID- 15824896 TI - The CT appearances of delayed amniotic fluid clearance from the lungs in an infant with absent pulmonary valve and congenital lobar emphysema. AB - Congenital lobar emphysema (CLE) is a cause of severe neonatal respiratory distress. Overexpansion of the affected pulmonary lobe in the fetus is due to narrowing of the airway, with a resultant 'ball-valve' effect. At birth, there may be delayed clearance of fetal lung fluid. Early chest radiographs show opacification of the hyperexpanded lobe. The CT findings in the immediate neonatal period have not been previously reported. We describe the imaging in a neonate with tetralogy of Fallot and absent pulmonary valve with secondary CLE. CT demonstrates the hyperexpanded lobe with initial thickening of the interlobular septa and alveolar ground glass attenuation, with subsequent clearing. This resorption of fetal lung fluid via the pulmonary interstitium should not be confused with interstitial lung disease. PMID- 15824897 TI - Molecular assessment of the elastic properties of collagen-like homotrimer sequences. AB - Knowledge of the mechanical behavior of collagen molecules is critical for understanding the mechanical properties of collagen fibrils that constitute the main architectural building block of a number of connective tissues. In this study, the elastic properties of four different type I collagen 30-residue long molecular sequences, were studied by performing stretching simulations using the molecular mechanics approach. The energy-molecular length relationship was achieved by means of the geometry optimization procedure for collagen molecule strains up to 10%. The energy was interpolated by a second order function, and the second order of the derivative with respect to the mean length corresponded to the molecule stiffness. According to the hypothesis of linear elastic behavior, except for one sequence, the elastic modulus was around 2.40 GPa. These values are larger than fibril values, and they confirm the hypothesis that tendon mechanical properties are deeply related to tendon hierarchical structure. A possible explanation of the lowest values obtained for one sequence (1.33-1.53 GPa) is provided and discussed. PMID- 15824898 TI - Locally recurrent rectal cancer: when should radiofrequency ablation be used? PMID- 15824899 TI - RNA interference screening in ticks for identification of protective antigens. AB - Ticks are ectoparasites of wild and domestic animals and humans, and are considered to be the most important arthropod vector of pathogens in North America. Development of vaccines directed against tick proteins may effect reduction of tick infestations and transmission of tick-borne pathogens. The limiting step for the development of tick vaccines has been the identification of tick protective antigens. Reverse vaccinology approaches aimed at reducing animal experimentation while allowing for the rapid screening of pools of potential tick vaccine candidates would greatly facilitate progress towards the development of tick vaccines. Herein, we describe the screening of Ixodes scapularis cDNAs for identification of tick protective antigens using RNA interference (RNAi). The results of the RNAi screening were similar to those obtained previously using expression library immunization and demonstrated that RNAi could serve as a more rapid and cost-effective tool for vaccine antigen discovery in ticks and in other nonmodel organisms. PMID- 15824900 TI - Seasonal prevalence and intensity of hookworms (Uncinaria spp.) in California sea lion (Zalophus californianus) pups born in 2002 on San Miguel Island, California. AB - Intestines of California sea lion (Zalophus californianus) pups (n= 204), born in 2002 on San Miguel Island, California, were examined for hookworms (Uncinaria spp.) as part of a seasonal mortality study from June through December 2002 and January 2003. The investigation was planned to coincide with most of the previously established hookworm infection period of the pups. Prevalence of hookworms in dead pups was 100% for each month of the study. The geometric mean intensity of infections per month was: 94.03 (n=30) for June, 629.09 (n=50) for July, 319.90 (n=31) for August, 159.90 (n=30) for October, 109.03 (n=30) for November, 37.84 (n=24) for December 2002 and 11.05 (n=9) for January 2003. In addition to the temporal pattern, the infection intensity was higher for pups in good condition and for male pups. An inter-year comparison of hookworm counts from dead pups collected in July of 1996, 2000, and 2002 also demonstrated higher intensity in pups in better condition but sex-differences in intensity were inconsistent across years. The inter-year comparison also demonstrated higher intensities in dead pups collected from portions of the rookery with sandy substrate versus rocky substrate. No annual differences in intensity were found after adjusting for substrate and condition. PMID- 15824901 TI - Domestic and wild mammals infection by Trypanosoma evansi in a pristine area of the Brazilian Pantanal region. AB - The objective of this study was to estimate the Trypanosoma evansi infection rate and epizootical status of wild and domestic animals from the Brazilian Pantanal region using a standardized polymerase chain reaction (PCR). We used a simple DNA extraction method based on Chelex resin (BioRad, USA) on blood eluted from filter paper confetti. Primers directed to repetitive nuclear DNA sequences were used in the PCR, and could detect 30 fg of T. evansi DNA. The analytical specificity of the assay was evaluated using T. evansi, T. rangeli, T. cruzi, Leishmania braziliensis, Crithidia fasciculata and Herpetomonas muscarum DNAs as templates and the technique showed the expected 164 bp specific band solely for Trypanozoon trypanosomes. The application of the standardized PCR protocol in 274 field samples from domestic and wild mammals from the Rio Negro (Brazilian Pantanal region), showed a general infection rate of 10.2% while the traditional parasitological technique (direct search of the protozoan by the microematocrit centrifugue technique) was able to determine infection in only 1.1% of the animals. The peccaries and feral pigs were found to be the animals most frequently infected with T. evansi (24.4% and 30.7%, respectively). Both sampling and extraction methods used herein, showed to be simple and efficient to be applied in epidemiological surveys using PCR. PMID- 15824902 TI - Immunogenicity of two Echinococcus granulosus antigens EgA31 and EgTrp in mice. AB - Here, we investigate in mice the immunogenicity of two antigens EgA31 and EgTrp which are expressed by the larval stage of Echinococcus granulosus. These recombinant proteins were used alone or as a mixture (EgA31-EgTrp) to immunize BALB/c mice. By flow cytometry, we have shown that the ratio CD4+/CD8+ of splenocytes were significantly higher in the antigen-immunized groups. The specific antibody in the sera and cytokine producing splenocytes was evaluated by enzyme-linked immunosorbent assay. EgA31, EgTrp or EgA31-EgTrp elicited high antibody titer of IgG and IgA. Among IgG isotypes, IgG1 was predominant for each antigen tested alone or combined. The production of IL-12, IFN-gamma, IL-10 and IL-6 cytokines was significantly higher in mice immunized with recombinant proteins. Our results suggest that, in BALB/c mice, a mixed Th1/Th2, response to EgA31, EgTrp and EgA31-EgTrp is obtained. The use of both antigens separately or in combination as candidate vaccine proteins is discussed. PMID- 15824903 TI - Molecular biological investigations of Brazilian Tunga sp. isolates from man, dogs, cats, pigs and rats. AB - Twenty-four isolates of Tunga were collected from afflicted humans, dogs, cats, pigs and rats in Brazil. To investigate genetic diversity, a hypervariable section of mitochondrial 16S rDNA was amplified using PCR and subsequently sequenced. In order to compare results with another species of the genus Tunga, three isolates of the recently described Tunga trimamillata were also checked. Whereas eleven isolates (five from cats, three from dogs and three from humans) were of identical sequence, thirteen isolates collected from dogs, humans, pigs and rats showed differences in sequence up to 49%, so that the existence of one or more new species of Tunga may be presumed. PMID- 15824904 TI - Development of genetic methods and construction of a chromosomal glnK1 mutant in Methanosarcina mazei strain Go1. AB - The methanogenic archaeon Methanosarcina mazei strain Go1 has so far proven to be genetically intractable due to its low plating efficiency on solid medium and the lack of an effective transformation method. Here, we report the first significant improvement in plating efficiency (up to 10%), which was achieved by (1) selecting for a spontaneous mutant of M. mazei that shows significantly higher resistance to mechanical stress during spreading an agar plates, and (2) plating the cells in 0.5% top agar with trimethylamine as a carbon and energy source under a H2S-containing atmosphere (0.1%). Using this mutant we succeeded in establishing a liposome-mediated transformation protocol, which for the first time allowed genetic manipulation of the M. mazei Go1 strain. We further report on the construction of the first chromosomal deletion mutant of M. mazei by means of homologous recombination. Characterization of this mutant strain revealed that M. mazei cells lacking a functional glnK1-gene exhibited a partial growth defect under nitrogen limitation when molecular nitrogen was used as the sole nitrogen source. Quantitative RT-PCR analysis, however, showed that genes involved in nitrogen assimilation or nitrogen fixation are transcribed in the glnK1 mutant as in the wild type. Thus, we propose that the archaeal GlnK1 protein is not directly involved in the transcriptional regulation of genes involved in nitrogen metabolism, but rather affects their protein products directly. PMID- 15824906 TI - Transgenic cotton (Gossypium hirsutum L.) seedlings expressing a tobacco glutathione S-transferase fail to provide improved stress tolerance. AB - Transgenic cotton (Gossypium hirsutum L.) lines expressing the tobacco glutathione S-transferase (GST) Nt107 were evaluated for tolerance to chilling, salinity, and herbicides, antioxidant enzyme activity, antioxidant compound levels, and lipid peroxidation. Although transgenic seedlings exhibited ten-fold and five-fold higher GST activity under normal and salt-stress conditions, respectively, germinating seedlings did not show improved tolerance to salinity, chilling conditions, or herbicides. Glutathione peroxidase (GPX) activity in transgenic seedlings was 30% to 60% higher under normal conditions, but was not different than GPX activity in wild-type seedlings under salt-stress conditions. Glutathione reductase, superoxide dismutase, ascorbate peroxidase, and monodehydroascorbate reductase activities were not increased in transgenic seedlings under salt-stress conditions, while dehydroascorbate reductase activity was decreased in transgenic seedlings under salt-stress conditions. Transgenic seedlings had 50% more oxidized glutathione when exposed to salt stress. Ascorbate levels were not increased in transgenic seedlings under salt-stress conditions. Malondialdehyde content in transgenic seedlings was nearly double that of wild-type seedlings under normal conditions and did not increase under salt-stress conditions. These results show that expression of Nt107 in cotton does not provide adequate protection against oxidative stress and suggests that the endogenous antioxidant system in cotton may be disrupted by the expression of the tobacco GST. PMID- 15824907 TI - Nitric oxide is involved in the Azospirillum brasilense-induced lateral root formation in tomato. AB - Azospirillum spp. is a well known plant-growth-promoting rhizobacterium. Azospirillum-inoculated plants have shown to display enhanced lateral root and root hair development. These promoting effects have been attributed mainly to the production of hormone-like substances. Nitric oxide (NO) has recently been described to act as a signal molecule in the hormonal cascade leading to root formation. However, data on the possible role of NO in free-living diazotrophs associated to plant roots, is unavailable. In this work, NO production by Azospirillum brasilense Sp245 was detected by electron paramagnetic resonance (6.4 nmol. g-1 of bacteria) and confirmed by the NO-specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA). The observed green fluorescence was significantly diminished by the addition of the specific NO scavenger 2-(4 carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). Azospirillum-inoculated and noninoculated tomato (Lycopersicon esculentum L.) roots were incubated with DAF-2 DA and examined by epifluorescence microscopy. Azospirillum-inoculated roots displayed higher fluorescence intensity which was located mainly at the vascular tissues and subepidermal cells of roots. The Azospirillum-mediated induction of lateral root formation (LRF) appears to be NO dependent since it was completely blocked by treatment with cPTIO, whereas the addition of the NO donor sodium nitroprusside partially reverted the inhibitory effect of cPTIO. Overall, the results strongly support the participation of NO in the Azospirillum-promoted LRF in tomato seedlings. PMID- 15824908 TI - Changes in cell wall polysaccharides in developing barley (Hordeum vulgare) coleoptiles. AB - Cell wall polysaccharides in developing barley coleoptiles were examined using acetic acid-nitric acid extraction, alditol acetate and methylation analyses and enzymatic digestion. The coleoptile cell wall from imbibed grain was rich in pectic polysaccharides (30 mol%), arabinoxylan (25 mol%), cellulose (25 mol%) and xyloglucan (6 mol%), but contained only low levels of (1-->3,1-->4)-beta-D-glucan (1 mol%). During 5 days of coleoptile growth, pectic polysaccharides decreased steadily to about 9 mol%, while (1-->3,1-->4)-beta-D-glucan increased to 10 mol%. Following the cessation of growth of the coleoptiles at about 5 days, (1-->3,1- >4)-beta-D-glucan content rapidly decreased to 1 mol%. The cellulose content of the walls remained at about 35-40 mol% throughout coleoptile growth. Similarly, arabinoxylan content remained essentially constant at 25-30 mol% during growth, although the ratio of substituted to unsubstituted 4-linked xylosyl units decreased from about 4:1 to 1:1. Xyloglucan content ranged from 6 mol% to 10 mol% and the oligosaccharide profile determined using a xyloglucan-specific endoglucanase and MALDI-TOF mass spectrometry indicated that the oligosaccharides XXGG and XXGGG were the principal components, with one and two acetyl groups, respectively, Thus, dramatic changes in wall composition were detected during the growth of barley coleoptiles, both with respect to the relative abundance of individual wall constituents and to the fine structure of the arabinoxylans. PMID- 15824909 TI - Computed tomography of the inner ear: size of anatomical structures in the normal temporal bone and in the temporal bone of patients with Meniere's disease. AB - The purpose of the study was to obtain reference values for the sizes of anatomical structures of the inner ear on computed tomography (CT) images and to compare these values with those obtained from patients with Meniere's disease. CT images of the temporal bone of 67 patients without inner ear pathology and 53 patients with Meniere's disease have been evaluated. CT was performed in the sequential mode (1-mm slice thickness, 120 kV, 125 mA). Anatomical structures, such as the length and the width of the cochlea and of the vestibule, the height of the basal turn, the length and the width of the cochlear, the vestibular and the singular aqueduct and the internal auditory meatus and the diameter of the semicircular canals, were measured, using a dedicated postprocessing workstation. Reference values from the control group could be obtained. In the patients with Meniere's disease, the length and the width of the vestibular aqueduct were smaller, compared with the values from the control group. The values obtained from the control group can serve as reference values for adult patients. The different sizes of anatomical structures of the control group and of patients suffering from Meniere's disease suggest that functional impairment might be related to subtle morphological changes. PMID- 15824910 TI - Underuse of lipid-lowering drugs and factors associated with poor adherence: a real practice analysis in Italy. AB - BACKGROUND: Many studies have indicated the adequate use of lipid-lowering drugs (LLDs) as a factor in reducing the risk of cardiovascular disease. However, in clinical practice, a very high percentage of patients are not adequately treated. OBJECTIVE: To analyze the management of hypercholesterolemia in a non experimental setting and to estimate the factors associated with poor adherence to treatment. METHODS: A longitudinal study was performed using clinical and demographic data recorded in the General Practitioners' database. The sample included all patients, aged 30 years or over, with total blood cholesterol measured between 1 January and 31 December 2000. Utilization of LLDs was defined as the standardized daily dose of the drugs purchased during the 12 months preceding the cholesterol measurement. RESULTS: The study included 4764 patients (mean age 59.4+/-14.1 years, 40.7% males). Of the subjects with a total cholesterol higher than a 6.5 mmol/l, approximately 17% were treated with LLDs. About 39% of the patients with previous atherosclerotic diseases were taking statins. Analysis of patients taking LLDs showed that 40.6% of subjects took less than half of the defined daily dose. Factors associated with poor adherence to treatment were: absence of previous atherosclerotic diseases, absence of concomitant diseases, and smoking. A total cholesterol of less than 5 mmol/l was achieved in 19.9% of patients. CONCLUSIONS: Analyzing the data contained in the general medicine database made it possible to evaluate the use of LLDs in clinical practice and to establish the need to pay greater attention to achieving the objective set by the treatment. PMID- 15824911 TI - Lower oropharyngeal deposition of inhaled ciclesonide via hydrofluoroalkane metered-dose inhaler compared with budesonide via chlorofluorocarbon metered-dose inhaler in healthy subjects. AB - OBJECTIVE: Inhaled corticosteroids may cause oropharyngeal side effects if deposited in the oropharynx in active form. Ciclesonide, an inhaled corticosteroid with low glucocorticoid receptor affinity, is activated primarily in the lung by esterases to an active metabolite, desisobutyryl-ciclesonide (des CIC), with high glucocorticoid receptor affinity. We studied oropharyngeal deposition of ciclesonide, des-CIC, and budesonide. METHODS: In an open-label, randomized, two-treatment (administered in sequence), five-period study, 18 healthy subjects received 800 microg (ex-valve) inhaled ciclesonide via a hydrofluoroalkane-pressurized, metered-dose inhaler followed by 800 microg budesonide (Pulmicort) by a chlorofluorocarbon-pressurized, metered-dose inhaler (four puffs of 200 microg each, ex-valve) or vice versa. Oropharyngeal cavity rinsing was performed immediately, or 15, 30, 45, or 60 min after inhalation (one rinsing per study period), and the solutions were analyzed using liquid chromatography with tandem mass spectrometric detection. RESULTS: Ciclesonide and budesonide were detected in most oropharyngeal wash samples. Maximal concentration of each inhaled corticosteroid was reached immediately post inhalation; maximal concentrations of ciclesonide and des-CIC were 30% and 0.67%, respectively, of budesonide. Oropharyngeal deposition of ciclesonide and budesonide decreased rapidly within 15 min post-inhalation, and less rapidly thereafter. Less than 10% of the residual ciclesonide in the oropharynx was converted to des-CIC. The molar dose-adjusted amount of des-CIC was 4% of budesonide (P < 0.0001). There were no significant adverse events. CONCLUSION: Oropharyngeal deposition of des-CIC was more than one order of magnitude lower than that of budesonide when administered by the respective metered-dose inhalers. This may explain the low frequency of oropharyngeal side effects of ciclesonide in clinical studies. PMID- 15824912 TI - Influence of cigarette smoking on melatonin levels in man. AB - OBJECTIVE: Polycyclic aromatic hydrocarbons in cigarette smoke induce cytochrome P450(CYP)1A2, which is involved in the hepatic metabolism of melatonin (MT). This suggests that habitual smokers may have low serum MT levels during smoking compared with a non-smoking period. We decided to investigate whether this suggestion is correct. METHODS: Eight habitual smokers were tested on two occasions. They had smoked prior to the first occasion but had not smoked for 7 days prior to the second. Each test was divided into two parts. The first part spanned the night between 2000 hours and 0800 hours. Venous blood samples were collected every second hour during this period for analysis of endogenous serum MT. The second part was performed the subsequent day. At 0930 hours, 25 mg MT was ingested orally, and blood samples for exogenous serum MT analysis were collected every hour between 1000 hours and 1600 hours. Endogenous and exogenous areas under the serum MT-time curve (MT-AUCs) were calculated. RESULTS: Endogenous serum MT-AUCs were similar during the two periods. Oral administration of MT induced supraphysiological levels of serum MT. Moreover, exogenous serum MT-AUCs were significantly smaller before than after smoking abstinence (7.34+/-1.85 versus 21.07+/-7.28 nmol/lxh; P<0.02; means+/-SEM). CONCLUSION: This investigation shows that exogenous, but not endogenous (at night), serum MT levels are influenced by cigarette smoking. When the MT levels are low, the influence of CYP1A2 levels appears to be less pronounced than when they are high, and the enzyme capacity hugely utilized. These findings implicate that interactions between exogenous MT, and substrates metabolized by CYP1A2, may differ in individuals before and after smoking abstinence. PMID- 15824913 TI - Interaction between mivacurium and pancuronium: impact of the order of administration. AB - INTRODUCTION: Potentiation of mivacurium by low-dose pancuronium is mostly due to an inhibition of plasma butyryl cholinesterase (BchE) resulting in a decreased rate of hydrolysis of mivacurium. Nevertheless, an interaction at the receptor site could not be ruled out. By changing the order of the muscle relaxant injections, we may lessen the pharmacokinetic interaction and assess the impact at the acetylcholine receptor level. METHODS: Twenty patients scheduled for general anesthesia with propofol and fentanyl, and isoflurane were randomized into two groups receiving, mivacurium 100 microg kg-1 followed by pancuronium 15 microg kg-1 (group 1) or pancuronium 15 microg kg-1 followed by mivacurium 100 microg kg-1 (group 2). BchE before and after injection of each relaxant was measured. Neuromuscular block was assessed with a force transducer at the adductor pollicis measuring the elicited twitch to ulnar nerve stimulation. RESULTS: The neuromuscular block was greater when pancuronium was administered before mivacurium (100% versus 96+/-3%; P<0.05). Times to recovery of the elicited twitch response to 25% and 75% of control value were increased by 100% (P<0.05). After pancuronium, decreases in BchE of 11% and 14% in groups 1 and 2 were observed, respectively. CONCLUSION: Interaction between mivacurium and low dose pancuronium is significant only when mivacurium is injected after pancuronium. PMID- 15824914 TI - Are incorrectly used drugs more frequently involved in adverse drug reactions? A prospective study. AB - OBJECTIVES: The aims of this study were to identify and evaluate the frequency of drugs involved in adverse drug reactions (ADRs) for which recommendations of the Summary of Product Characteristics (SPC) had not been respected and to compare the percentages of drugs suspected of being the cause of ADRs when correctly and incorrectly used. METHODS: All ADRs reported to the Regional Pharmacovigilance Centre of Tours (RPC) over a period of 5 months were analysed to identify drugs used "incorrectly" defined by drug use beyond the recommendations of the SPC, i.e. drugs used when contra-indicated and/or drugs used for an off-label indication, and/or drugs used at an inappropriate dose, and/or inappropriate duration of treatment, and/or drugs used in the presence of a potentially or definitely interacting drug. RESULTS: Included in the study were 182 ADRs involving 182 patients. Of 642 drugs, 169 (26%) were incorrectly used and for 81 patients (44.5%) ADRs involved at least one "incorrectly" used drug. These included 10% (64 of 642) drug interactions, 7.3% (47 of 642) off-licence indications, 5% (32 of 642) inadequate dosage, 3% (20 of 642) incorrect duration of treatment and 1% (6 of 642) contraindications. "Correctly" used drugs appeared to be less often the cause of the ADRs than "incorrectly" used drugs (59.4% versus 75%, P=0.0001). CONCLUSION: "Incorrectly" used drugs were more often causally linked to ADR than correctly used drugs. A meaningful number of ADRs could probably be avoided if SPC guidelines for a safe and effective drug use are carefully adhered to. PMID- 15824915 TI - Effects of shift work on QTc interval and blood pressure in relation to heart rate variability. AB - OBJECTIVES: There is evidence that shift work contributes to excess cardiovascular mortality. The purpose of this study was to determine the effects of shift work on heart rate-corrected QT (QTc) and blood pressure in relation to heart rate variability (CV(RR)). METHODS: The study population consisted of 153 male shiftworkers and 87 male day workers who were employed at a copper-smelting plant. The QTc interval, total power spectral density (t-PSD) of 100 RR intervals, PSDs with frequencies of 0.01 Hz-0.15 Hz and 0.15 Hz-0.40 Hz (PSD(LF) and PSD(HF)), CV(RR), low frequency (LF) component variability and high frequency (HF) component variability (CCV(LF) and CCV(HF)) and %LF (PSD(LF)/(PSD(LF)+PSD(HF)) .100) were measured (LF and HF components are thought to reflect the sympathetic and parasympathetic activities, respectively). RESULTS: The QTc interval was significantly longer in the shiftworkers than in the day workers, although there was no significant difference in systolic or diastolic blood pressure between the two groups. Also, the CCV(LF) and log(PSD(LF)) were significantly depressed in the shiftworkers. In the day workers, the QTc interval was significantly related to the CV(RR), log(t-PSD), CCV(HF), log(PSD(HF)),%LF, and log(LF/HF ratio) (partial correlation coefficient r=-0.305, -0.377, -0.312, -0.355, 0.297, and 0.277, respectively). In the shiftworkers only two relations of the QTc interval to non-specific CV(RR) and log(t-PSD) were significant. CONCLUSION: The clear association between long QTc interval and reduced parasympathetic activity observed in the day workers did not exist in the shiftworkers. Rather, the shiftworkers had a selective reduction in sympathetic drive, which may have compensated for prolonged QTc interval due to shift work. On the other hand, the impact of shift work on hypertension remains unclear, although reduced sympathetic drive in the shiftworkers may explain the absence of differences in blood pressure between shiftworkers and day workers to some extent. PMID- 15824917 TI - An overview of the Sino-German symposium on new developments in surgical and basic pathology. PMID- 15824916 TI - Workplace stress, lifestyle and social factors as correlates of back pain: a representative study of the German working population. AB - OBJECTIVES: To investigate the prevalence of back pain in the German working population and the relationship between back pain and workplace stresses, lifestyle and social factors. METHODS: The first National Health Survey of the Federal Republic of Germany was carried out between October 1997 and March 1999. It comprised a representative epidemiological cross-sectional study of the working population, with a total sample of 3,488 persons between the ages of 18 and 69 years. The participants took part in a medical examination and answered a self-rating questionnaire. The relationship between subjective back pain and workplace stresses and social and lifestyle factors was investigated with bivariate tests and multiple logistical regression analyses. RESULTS: The 7-day prevalence for back pain in the German working population was found to be 34%, and the 1-year prevalence was 60%. The odds ratios were significantly higher in women, persons of lower socioeconomic status, married and depressed persons and non-athletes. Carrying heavy loads or maintaining a single working posture were the most significant work-related correlates of back pain, for members of both the female and male working population, while environmental stress and psychological stress correlated significantly with back pain in men only. CONCLUSIONS: This study reports the first representative epidemiological prevalence data for back pain, and its correlates and potential risk factors, for the German working population. To reduce the negative impact of back pain the most promising behavioural and conditional prevention measures in the workplace would be to reduce carrying stress and to vary working posture. In addition, a more active, athletic lifestyle, plus the avoidance of being overweight, should provide an additional protective or preventive effect. PMID- 15824918 TI - Design, synthesis, physical and chemical characterisation, and biological interactions of lectin-targeted latex nanoparticles bearing Gd-DTPA chelates: an exploration of magnetic resonance molecular imaging (MRMI). AB - The physical and chemical parameters involved in the design and synthesis of biospecifically targeted nanoparticulate contrast media for magnetic resonance molecular imaging (MRMI) were explored in this pilot investigation. Latex nanoparticles 100, 400 and 900 nm in diameter were doubly derivatised, first with tomato lectin and then with gadolinium(III)-diethylenetriamine pentaacetic acid (Gd-chelates) to target them to epithelial and endothelial glycocalyceal N glycans and to generate contrast enhancement in magnetic resonance imaging (MRI). After intravenous injection into mice, human placental cotyledons or human Vena saphena magna, contrasty images of the vascular structures were obtained in 1.5 T MRI with spatial resolution 0.1 mm in the imaging plane and 0.6 mm in the z axis, persisting >60 min and resistant to washing out by buffer rinses. Ultrastructural analysis of the nanoparticles revealed the targeting groups at the nanoparticle surfaces and the distribution of the Gd-chelates within the nanoparticles and enabled counts for use in determining relaxivity. The relaxivity values revealed were extremely high, accounting for the strong MR signals observed. Occasionally, nanoparticles larger than 100 nm were seen in close spatial association with disrupted regions of cell membrane or of collagen fibrils in the extracellular matrix. The data suggest that 100-nm nanoparticles generate adequate contrast for MRMI and cause least disruption to endothelial cell surfaces. PMID- 15824921 TI - Guidelines for the pharmacotherapy of premature ejaculation. AB - In the 1960s, premature ejaculation was considered a learned behaviour that had to be treated by behavioural therapy, such as the so-called stop-start technique. The efficacy of this method was questioned. Physicians have always looked for medication that could delay ejaculation. Many studies, controlled and open label, have now clearly shown that rapid ejaculation can be treated quickly and effectively with clomipramine and SSRIs. This article describes the guidelines for the pharmacological treatment of premature ejaculation. These guidelines were developed to ensure that medication are used when clear indications are present. PMID- 15824922 TI - The solvent-tolerant Pseudomonas putida S12 as host for the production of cinnamic acid from glucose. AB - A Pseudomonas putida S12 strain was constructed that efficiently produced the fine chemical cinnamic acid from glucose or glycerol via the central metabolite phenylalanine. The gene encoding phenylalanine ammonia lyase from the yeast Rhodosporidium toruloides was introduced. Phenylalanine availability was the main bottleneck in cinnamic acid production, which could not be overcome by the overexpressing enzymes of the phenylalanine biosynthesis pathway. A successful approach in abolishing this limitation was the generation of a bank of random mutants and selection on the toxic phenylalanine anti-metabolite m-fluoro phenylalanine. Following high-throughput screening, a mutant strain was obtained that, under optimised culture conditions, accumulated over 5 mM of cinnamic acid with a yield (Cmol%) of 6.7%. PMID- 15824923 TI - Inhibition of P-glycoprotein transport function and reversion of MDR1 multidrug resistance by cnidiadin. AB - PURPOSE: Overexpression of P-glycoprotein (Pgp) encoded by the MDR1 gene is one of the major obstacles to successful cancer chemotherapy. The goal of this study was to evaluate if, among other natural coumarins, cnidiadin, a furanocoumarin present in traditional Chinese medications and in a spice commonly used in Greek food, inhibits Pgp transport activity and has the potential to reverse MDR1 multidrug resistance. METHODS: Using MDR1-transfected Madin-Darby canine kidney (MDCK-MDR1) cells as a model of cells expressing the human MDR1 phenotype, and verapamil or CsA or both as positive control, we tested the capacity of six natural coumarins (umbelliferone, esculin, esculetin, cnidiadin, angelicin and psoralen) to induce the accumulation of rhodamine-123 (R-123) and [3H] vinblastine ([3H]-VBL) and to modulate the photolabeling of Pgp by SDZ 212-122, a diazirin cyclosporin A. The growth-inhibitory effect of cnidiadin and its capacity to enhance the cell toxicity of vinblastine (VBL) or vincristine (VCR) was then evaluated by the WST-1 assay in two cell lines overexpressing Pgp (MDCK MDR1 and vincristine-resistant KB/VCR). RESULTS: Cnidiadin was the only tested coumarin capable of significantly accumulating R-123 and [3H]-VBL and inhibiting Pgp photolabeling in MDCK-MDR1 cells. The dose-dependent increase in [3H]-VBL uptake (IC50 26.5 microM) induced by cnidiadin in the dose range 1-100 microM correlated with inhibition of Pgp photolabeling. At 10 microM cnidiadin inhibited photolabeling by 59% and sensitized both MDCK-MDR1 and KB/VCR cells to vinca alkaloids. CONCLUSION: Cnidiadin is a cytotoxic agent capable in vitro of competitively inhibiting the binding and efflux of drug by Pgp and of enhancing the cell toxicity of vinca alkaloids in two cell lines (MDCK-MDR1 and mutant human carcinoma KB/VCR) overexpressing Pgp. This suggests that diet or traditional preparation containing cnidiadin may contribute to the reversal of MDR1 multidrug resistance and may affect the bioavailability of Pgp substrates orally administered. However, due to its cell toxicity, clinical interest in cnidiadin as a chemosensitizer appears to be limited. PMID- 15824924 TI - Dinuclear platinum anticancer complexes with fluorescent N,N'-bis(aminoalkyl)-1,4 diaminoanthraquinones: cellular processing in two cisplatin-resistant cell lines reflects the differences in their resistance profiles. AB - The biological activity of N,N'-bis(aminoalkyl)-1,4-diaminoanthraquinones (aminoalkyl is 2-aminoethyl, 3-aminoprop-1-yl and 4-aminobut-1-yl) and their dinuclear platinum complexes has been evaluated in the U2-OS human osteosarcoma cell line and its cisplatin-resistant U2-OS/Pt subline. All the compounds have been found to exhibit high cytotoxicity in the sensitive cell line, and to overcome cisplatin resistance in U2-OS/Pt cells. Cellular processing of N,N' bis(2-aminoethyl)-1,4-diaminoanthraquinone and the respective dinuclear platinum complex in the sensitive and resistant U2-OS cells has been studied over time using digital fluorescence microscopy. Cellular processing of the compounds has been found to be similar in sensitive and resistant U2-OS cells, which is in agreement with the lack of cross-resistance in the U2-OS/Pt cell line. Both the platinum complex and the free ligand quickly enter the cell and accumulate in the nucleus. The platinum complex is excreted from the cell via the Golgi apparatus, while the weakly basic anthraquinone ligand accumulates in the Golgi complex, where it is taken up by lysosomes and then transported to the cell surface. The cellular distribution of the fluorescent anthraquinones and their dinuclear platinum complexes in the sensitive/resistant pair of U2-OS osteosarcoma cell lines is compared with the earlier studied cellular processing in the sensitive/resistant pair of A2780 ovarian carcinoma cell lines. In the A2780cisR cell line, the platinum complexes (and not the free ligands) are sequestered in lysosomes, which is not the case in A2780 sensitive cells. The differences in cellular distribution of the compounds in these two sensitive/resistant pairs of cell lines most likely result from different resistance profiles in A2780cisR and U2-OS/Pt cells. Lysosomes of A2780cisR cells are less acidic than lysosomes of A2780 sensitive cells, which is likely to be the cause of a defect in endocytosis. The disruption of normal endocytosis might facilitate sequestration of the platinum complexes in lysosomes, which partly confers the cross-resistance of these complexes with cisplatin in the A2780cisR cell line. In contrast, sequestration in acidic vesicles does not occur in U2-OS/Pt cells that do not exhibit enhanced lysosomal pH and which are likely to have normal endocytosis. PMID- 15824925 TI - Use of myocardial perfusion imaging to predict the effectiveness of coronary revascularisation in patients with stable angina pectoris. AB - PURPOSE: Coronary revascularisation is the treatment of choice in patients with stable angina who have significant stenoses. From a pathophysiological point of view, however, mitigation of angina is to be expected only in the presence of reversible ischaemia. Therefore it was the aim of this study to examine the effect of revascularisation on stable angina in relation to the myocardial perfusion imaging (MPI) pattern prior to intervention. METHODS: Three hundred and eighty-four patients (58.0+/-8.8 years) referred for angiography underwent MPI. Prior to MPI and at 2-year follow-up, patients were classified as having typical angina, atypical angina, non-cardiac chest pain or no pain, and the severity of chest pain was graded according to the Canadian Cardiovascular Society (CCS) criteria. The patients themselves estimated their pain on a visual analogue scale. Management was based on symptoms and angiographic findings, since the results of MPI were not communicated. RESULTS: Among the 240 patients who were not revascularised, 79% had typical or atypical angina at study entrance versus 40% at follow-up. In comparison, 93% of the 144 revascularised patients had typical or atypical angina before intervention versus only 36% at follow-up. This additional advantage of invasive therapy was present only in patients with reversible defects; revascularisation had no additional effect in patients with normal perfusion or irreversible defects. Similarly, additional, significant reductions in CCS class and visual analogue score were observed exclusively in patients with reversible defects. CONCLUSION: In patients referred for coronary angiography owing to known or suspected stable angina, revascularisation was significantly more effective than medical treatment exclusively in patients with reversible ischaemia. PMID- 15824926 TI - Repeated injections of 131I-rituximab show patient-specific stable biodistribution and tissue kinetics. AB - PURPOSE: It is generally assumed that the biodistribution and pharmacokinetics of radiolabelled antibodies remain similar between dosimetric and therapeutic injections in radioimmunotherapy. However, circulation half-lives of unlabelled rituximab have been reported to increase progressively after the weekly injections of standard therapy doses. The aim of this study was to evaluate the evolution of the pharmacokinetics of repeated 131I-rituximab injections during treatment with unlabelled rituximab in patients with non-Hodgkin's lymphoma (NHL). METHODS: Patients received standard weekly therapy with rituximab (375 mg/m2) for 4 weeks and a fifth injection at 7 or 8 weeks. Each patient had three additional injections of 185 MBq 131I-rituximab in either treatment weeks 1, 3 and 7 (two patients) or weeks 2, 4 and 8 (two patients). The 12 radiolabelled antibody injections were followed by three whole-body (WB) scintigraphic studies during 1 week and blood sampling on the same occasions. Additional WB scans were performed after 2 and 4 weeks post 131I-rituximab injection prior to the second and third injections, respectively. RESULTS: A single exponential radioactivity decrease for WB, liver, spleen, kidneys and heart was observed. Biodistribution and half-lives were patient specific, and without significant change after the second or third injection compared with the first one. Blood T(1/2)beta, calculated from the sequential blood samples and fitted to a bi-exponential curve, was similar to the T(1/2) of heart and liver but shorter than that of WB and kidneys. Effective radiation dose calculated from attenuation-corrected WB scans and blood using Mirdose3.1 was 0.53+0.05 mSv/MBq (range 0.48-0.59 mSv/MBq). Radiation dose was highest for spleen and kidneys, followed by heart and liver. CONCLUSION: These results show that the biodistribution and tissue kinetics of 131I-rituximab, while specific to each patient, remained constant during unlabelled antibody therapy. RIT radiation doses can therefore be reliably extrapolated from a preceding dosimetry study. PMID- 15824927 TI - Comparison of 99mTc-sestamibi/18FDG DISA SPECT with PET for the detection of viability in patients with coronary artery disease and left ventricular dysfunction. AB - PURPOSE: Dual-isotope simultaneous acquisition (DISA) single-photon emission computed tomography (SPECT) is an attractive technique as it permits assessment of both myocardial glucose metabolism and perfusion within a single session, but few data on its accuracy for the assessment of viability are available as yet. In the present study, DISA SPECT was compared with positron emission tomography (PET) for the detection of myocardial viability in normal and dysfunctional left ventricular (LV) myocardium. METHODS: Fifty-eight patients with chronic coronary artery disease and LV dysfunction (LV ejection fraction 33+/-12%) were studied. Patients underwent a 1-day dipyridamole stress 99mTc-sestamibi/18F fluorodeoxyglucose (18FDG) DISA SPECT and 13N-ammonia/18FDG PET protocol. Within 1 week, resting MRI was performed to assess contractile function. Comparison of PET and SPECT data was performed using both visual and quantitative analysis. RESULTS: The correlation of normalised activities of the flow tracers 99mTc sestamibi and 13N-ammonia was good (r = 0.82; p < 0.001). The correlation between the two 18FDG studies was also good (r = 0.83; p < 0.001). The agreement for the assessment of viability for all segments between DISA SPECT and PET was 82%, with a kappa-statistic of 0.59 (95% CI 0.53-0.64), without a significant difference; in dysfunctional segments only, the agreement was 82%, with a kappa-statistic of 0.63 (95% CI 0.56-0.70), without a significant difference. When the DISA SPECT data were analysed visually, the agreement between DISA SPECT and PET was 83%, with a kappa-statistic of 0.58 (95% CI 0.52-0.63), without a significant difference. Moreover, there was no significant difference between visual and quantitative DISA SPECT analysis for the detection of viability. CONCLUSION: This study shows an overall good agreement between 99mTc-sestamibi/18FDG DISA SPECT and PET for the assessment of myocardial viability in patients with severe LV dysfunction. Quantitative or visual analysis of the SPECT data did not influence the agreement with PET, suggesting that visual assessment may be sufficient for clinical purposes. PMID- 15824929 TI - [Carl Freiherr von Rokitansky: a commemoration of his 200th birthday]. PMID- 15824928 TI - Littre's hernia in newborn infants: report of two cases. AB - Littre's hernia was originally defined by Reinke in 1841 as "the presence of a Meckel's diverticulum in any hernial sac." It is generally difficult to differentiate from other types of hernia until complications arise. It is a rare and accidental finding at any age, but it is absolutely exceptional in neonates. To our knowledge, only one case under the age of 1 month has been reported in the literature. Herein two cases treated in the first 20 days of life are reported. PMID- 15824930 TI - Locomotion and the pollen hoarding behavioral syndrome of the honeybee (Apis mellifera L.). AB - Honeybees selected for the colony level phenotype of storing large quantities of pollen (pollen hoarding) in the nest exhibit greater walking activity than those selected against pollen hoarding. In this study, we use a simple walking assay to demonstrate that walking activity increases with the proportion of high pollen hoarding alleles in pure and backcrossed strains of bees (high-strain bees > offspring generated from a high backcross > offspring generated from a low backcross > low-strain bees). The trait is heritable but is not associated with markers linked to three quantitative trait loci (QTL) mapped for their effects on pollen hoarding with demonstrated pleiotropic effects on pollen and nectar foraging and learning behavior. However, locomotion in non-selected bees is correlated with responsiveness to sucrose, a trait that correlates with foraging and learning behavior. We propose that pollen-hoarding behavior involves a syndrome of behavioral traits with complex genetic and regulatory architectures that span sensory sensitivity, foraging behavior, and learning. We propose that locomotor activity is the component of this syndrome and reflects the early maturation of the bees that become pollen foragers. PMID- 15824932 TI - The role of pre-operative templating in primary total knee replacement. AB - Templating of preoperative X-rays is routinely performed before a total knee replacement. We performed this retrospective study on preoperative templating in primary total knee replacement to assess its accuracy and reproducibility. Preoperative radiographs of 47 patients were templated twice by two observers separately after a gap of 3 months. The size of actual implant used was compared to the size predicted by templating. Data were analysed to assess inter and intra observer variation. Templating was accurate for both tibial and femoral components in only 53.2% of observations. This study also revealed that templating is a highly-subjective and observer-dependent technique. Inter observer and intra-observer mismatch was present in 46.8% and 43.6% of readings respectively. We conclude from this study that preoperative templating is neither accurate nor reproducible. PMID- 15824933 TI - Acute locking in revision total knee arthroplasty due to disengagement of the locking screw. AB - Acute locking of the joint in a replaced knee joint is very rare. This report describes an acute locking episode of a revised modular total knee replacement, occurring more than 2 years after surgery. A disengaged screw from the modular femoral component had lodged in the joint at the inferior pole of the patella and required urgent arthroscopic removal. There was no subsequent failure of the stem condylar junction, nor loosening of the femoral component. PMID- 15824934 TI - The influence of tibial slope on maximal flexion after total knee arthroplasty. AB - Many surgeons believe that increasing the tibial slope in total knee arthroplasty (TKA) is beneficial with regard to maximal postoperative flexion. Review of the clinical literature, however, does not confirm this hypothesis, neither does it give an answer to the question of how much flexion gain can be expected per degree extra tibial slope. The purpose of this study was, therefore, to evaluate and quantify the influence of tibial slope on maximal postoperative flexion in contemporary posterior cruciate ligament (PCL)-retaining TKA. Twenty-one cadaver simulations of a standard PCL-retaining TKA were studied while reproducing identical deep flexion femorotibial kinematics as documented by three-dimensional computer-aided videofluoroscopy from patients with well-functioning TKAs of the same design. In each knee the tibial component was consecutively implanted with 0 degrees posterior slope, 4 degrees posterior slope, and 7 degrees posterior slope. Maximal flexion was recorded for each configuration. Average maximal flexion at 0 degrees tibial slope was 104 degrees, and increased significantly to 112 degrees when the same knees were implanted with 4 degrees tibial slope. Increasing the slope further to 7 degrees again significantly improved average maximal flexion to 120 degrees. When postoperative radiographic tibial slope was compared to maximal flexion, an average gain of 1.7 degrees flexion for every degree extra tibial slope was noted. Increasing the tibial slope in PCL-retaining TKA does indeed improve maximal flexion before tibial insert impingement occurs against the femoral bone. The surgeon can expect an average gain of 1.7 degrees flexion for every degree extra tibial slope. PMID- 15824935 TI - Anatomical repair of lateral ligaments in patients with chronic ankle instability. AB - In a prospective study, 19 patients with chronic ankle instability underwent clinical and radiographic reexaminations 36 months after anatomical reconstruction. In addition, dynamic pedography was conducted and peroneal reaction time measured on a tilting platform for an evaluation of functional aspects. Prior to this examination, 32 patients had been asked to fill in a questionnaire and make a detailed subjective evaluation of current discomfort, stability, flexibility and sporting abilities. Eighty-eight percent of the patients reported satisfactory results; only 3% complained of persistent instability. In 71% the ability to take part in sports had improved after surgery, and 85% of the patients reported unrestricted walking abilities. Supination ability was impaired in 5% of the patients at the follow-up. The radiographic examination showed restored ankle stability with a significant reduction of talar tilt and talar translation; a postoperative increase in signs and symptoms of arthrosis was not observed. Dynamic pedography showed a large degree of symmetry of plantar pressure distribution after surgery. There were no significant differences in peroneal reaction time in the repaired and intact ankles. The results of the study show that it is possible to restore ankle stability with anatomical reconstruction without impairing the range of movement in the ankle joint complex. Progressive osteoarthrosis can be prevented. PMID- 15824936 TI - Bioconversion of phytosterols to androstanes by mycobacteria growing on sugar cane mud. AB - Direct sterol conversion of sugar cane mud (residue) by Mycobacterium sp. was demonstrated to be possible technologically, thus avoiding sugar cane oil extraction and further processes of extraction and purification of phytosterols from this oil. Indeed, mycobacterial cells were able to convert phytosterols from sugar cane mud into 4-androstene-dione (AD) and 1,4 androsta-diene-3,17-dione (ADD). For the various concentrations assayed, concomitant higher yields for both androstanes were achieved at 20% (w/w) sugar cane mud in media. Furthermore, conversions were similar to those from other substrates, such as a mixture of phytosterols. The results suggest that the mycobacterial cell is able to easily access and bioconvert sugar cane mud phytosterols. PMID- 15824937 TI - EGGS for patient-centered outcomes. PMID- 15824938 TI - Responses to a simple barter task in chimpanzees, Pan troglodytes. AB - Chimpanzees (Pan troglodytes) frequently participate in social exchange involving multiple goods and services of variable value, yet they have not been tested in a formalized situation to see whether they can barter using multiple tokens and rewards. We set up a simple barter economy with two tokens and two associated rewards and tested chimpanzees on their ability to obtain rewards by returning the matching token in situations in which their access to tokens was unlimited or limited. Chimpanzees easily learned to associate value with the tokens, as expected, and did barter, but followed a simple strategy of favoring the higher value token, regardless of the reward proffered, instead of a more complex but more effective strategy of returning the token that matched the reward. This response is similar to that shown by capuchin monkeys in our previous study. We speculate that this response, while not ideal, may be sufficient to allow for stability of the social exchange system in these primates, and that the importance of social barter to both species may have led to this convergence of strategies. PMID- 15824939 TI - Endoscopic gastroenteric anastomosis using magnets. AB - BACKGROUND: Current management of malignant obstruction of the upper digestive tract includes surgical gastrointestinal bypass or endoscopic insertion of self expandable metal stents. The safety, efficacy, and long-term patency rates of anastomoses created using the novel technique of endoscopic gastroenteric anastomosis using magnets (EGAM) are evaluated in this study. PATIENTS AND METHODS: 15 patients (13 men, 2 women; mean age 64.5 years) with malignant obstruction, who underwent EGAM and had monthly follow-up between December 2001 and May 2003, were included in this study. RESULTS: The procedure was successful in 13 patients (88.66 %). The mean survival was 5.23 months. There were four minor complications (30.76 %) during the follow-up period. CONCLUSION: Our results demonstrate the feasibility, safety. and efficacy of this technique for creating a gastroenteric anastomosis. The success rate was 86.6 %, there were no immediate complications, and there was no mortality related to the procedure. PMID- 15824940 TI - Clinical impact of capsule endoscopy compared to push enteroscopy: 1-year follow up study. AB - BACKGROUND AND STUDY AIMS: The long-term outcome for patients with obscure bleeding after capsule endoscopy (CE) is still unclear. In this study, the clinical outcome was used as the gold standard to determine the sensitivity and specificity of CE and push enteroscopy (PE) in the diagnosis of small-bowel lesions in patients with obscure bleeding. PATIENTS AND METHODS: Fifty-eight patients from a previous prospective study (comparing PE and CE) were included; the patients were contacted after 1 year. The final diagnosis, bleeding status, new gastrointestinal examinations, and treatments performed were recorded. On the basis of these data, each case was classified into true/false positive or true/false negative findings at PE and CE. The results were compared with the initial classification of lesions observed at CE: highly relevant (P2) and less relevant (P0, P1) lesions. RESULTS: Follow-up data were available for 56 patients. According to the defined true/false positive and negative cases, the sensitivity and specificity values for CE and PE were 92 % and 48 %, and 80 % and 69 %, respectively ( P < 0.01 for the difference between CE and PE). Highly relevant (P2) lesions observed at CE were more frequently classified into true positive cases (15 of 18 versus seven of 22; P < 0.01) and led more frequently to therapeutic decisions (11 of 18 versus five of 22; P = 0.02) in comparison with less relevant lesions (P0, P1). CONCLUSIONS: CE is a highly sensitive examination for the detection of small-bowel lesions in patients with obscure gastrointestinal bleeding, with a specificity lower than that of PE when the clinical outcome is used as the gold standard. PMID- 15824941 TI - Is early endoscopy in the emergency room beneficial in patients with bleeding peptic ulcer? A "fortuitously controlled" study. AB - BACKGROUND AND STUDY AIMS: In previous randomized trials, early endoscopy improved the outcome in patients with bleeding peptic ulcer, though most of these studies defined "early" as endoscopy performed within 24 hours after admission. Using the length of hospital stay as the primary criterion for the clinical outcome, we compared the results of endoscopy done immediately after admission (early endoscopy in the emergency room, EEE) with endoscopy postponed to a time within the first 24 hours after hospitalization, but still during normal working hours ("delayed" endoscopy in the endoscopy unit, DEU). PATIENTS AND METHODS: We conducted a retrospective analysis of data from 81 consecutive patients with bleeding peptic ulcer admitted in 1997 and 1998 (age range 16 - 90 years). Of these 81 patients, 38 underwent DEU (the standard therapy at the hospital) and 43 underwent EEE. Patients in the two groups were comparable with regard to admission criteria, were equally distributed with respect to their risk of adverse outcome (assessed using the Baylor bleeding score and the Rockall score), and differed only in the treatment they received. Endoscopic hemostasis was performed whenever possible in all patients with Forrest types I, IIa, and IIb ulcer bleeding. RESULTS: We found similar rates in the two groups for recurrent bleeding (16 % in DEU patients vs. 14 % in EEE patients), persistent bleeding (8 % in DEU patients vs. none in EEE patients), medical complications (21 % in DEU patients vs. 26 % in EEE patients), the need for surgery (8 % in DEU patients vs. 9 % in EEE patients), and the length of hospital stay (5.1 days for DEU patients vs. 5.9 days for EEE patients). None of the differences between the two groups in these parameters were statistically significant. None of the patients died. CONCLUSIONS: Early endoscopy in an emergency room did not improve the clinical outcome in our 81 consecutive patients with bleeding peptic ulcer. PMID- 15824942 TI - A nationwide study of the use of self-expanding stents in patients with esophageal cancer in Sweden. AB - BACKGROUND AND STUDY AIMS: Endoscopic insertion of self-expanding metal stents is used for palliative treatment of esophageal and cardia cancer, but the safety profiles, and the influence of age, sex, and hospital volume have not previously been evaluated in a population-based setting. PATIENTS AND METHODS: A retrospective population-based study was conducted, including all patients treated with esophageal stenting for malignant disease recorded in Swedish national registries in the period 1997 - 2000. Patients with esophageal or cardia cancer were identified in the cancer registry, and those treated with esophageal stenting were selected from the in-patient registry. Survival and emigration were assessed using the registries for causes of death and emigration, respectively. Procedure-related complications were assessed by reviewing medical records. RESULTS: Among 1052 registered patients with esophageal or cardia cancer, 402 patients (38 %), with a median age of 74 years, were treated with esophageal stenting. After treatment, the median in-hospital stay was 7 days, and the median survival was 100 days. Survival times were similar between age groups and sexes. Stenting was conducted at 38 Swedish hospitals, with a range of 1-59 procedures at each hospital. Among 152 (38 %) patients who were evaluated for complications, 41 (27 %) had complications of some type, while 20 (13 %) experienced direct procedure-related complications. No differences in complication frequencies were found between high-volume and low-volume centers. There were only two procedure related deaths (1 %), both due to esophageal perforation. No reduction in the median survival time was found in patients with complications. CONCLUSIONS: Treatment with self-expanding metal stents is a widely used and reasonably safe procedure among patients with malignant dysphagia, independent of sex, age, or hospital volume in Sweden. PMID- 15824943 TI - Management of malignant stricture of the esophagogastric junction with a newly designed self-expanding metal stent with an antireflux mechanism. AB - BACKGROUND AND STUDY AIMS: When stents are placed across the esophagogastric junction for palliative treatment of malignant strictures, they may lead to esophagogastric reflux. The aim of this study was to compare the effectiveness of a newly designed antireflux stent with that of a standard open stent and a currently available antireflux stent (Dostent) in preventing gastroesophageal reflux symptoms in patients with inoperable cancer at the esophagogastric junction. PATIENTS AND METHODS: Thirty-six consecutive patients with cancer at the esophagogastric junction were randomly assigned to undergo placement of a newly designed antireflux stent (n = 12), a Dostent (n = 12), or a standard open stent (n = 12). Technical and clinical success, dysphagia score, reflux symptoms, complications and ambulatory 24-h esophageal pH monitoring were assessed. RESULTS: The technical success rates were 100 %. After 1 week, dysphagia had improved in all patient groups ( P < 0.05), but the degree of improvement did not differ between the three groups. The DeMeester score was significantly lower in the group with the newly designed antireflux stent than in the other groups. The fraction of the total recording time during which esophageal pH was below 4 was 3.14 +/- 5.78 % using the newly designed antireflux stent, in comparison with 29.25 +/- 15.41 % in the Dostent group and 15.01 +/- 11.72 % in the standard open stent group ( P < 0.001). Fewer reflux episodes occurred with the newly designed antireflux stent than with the Dostent or standard open stent. There were no complications with any of the three stents. CONCLUSIONS: The newly designed antireflux stent is effective in relieving dysphagia caused by malignant cancer at the esophagogastric junction. The newly designed antireflux stent is significantly more effective in preventing gastroesophageal reflux than currently available antireflux stents. PMID- 15824944 TI - Oil-lubricated colonoscopy: easier and less painful? AB - BACKGROUND AND STUDY AIMS: Methods of lubrication are rarely considered to be a matter for study in gastrointestinal endoscopy. We evaluated a new technique, i. e. the release of seed oil in discrete amounts from the tip of the scope during colonoscopy. PATIENTS AND METHODS: 346 consecutive patients prospectively underwent colonoscopy with a standard lubricating method (using water-soluble jelly; group A) or with the standard method plus seed oil (corn oil) instillations through the biopsy channel (group B). The following variables were evaluated in the two groups: the success rate for total intubation, and the time required to reach the cecum; the time needed to examine the colon at withdrawal; the detection rates for colorectal diseases; the level of pain and degree of difficulty associated with the examination. Patients in whom total colonoscopy was not achieved were asked to undergo a further examination in which the other lubricating technique was used. RESULTS: Successful intubation to the cecum was significantly more frequent ( P < 0.005) in the oil lubrication group (group B, 159/168) than in the control group (group A, 145/170), and less time was needed ( P < 0.001). No significant differences were found with regard to time for examination at withdrawal and detection rates for colorectal diseases. Level of pain and degree of difficulty during colonoscopy were significantly lower in the oil group ( P < 0.001). In the cross-over examinations done in patients in whom total colonoscopy was not achieved, no statistical difference was found between the two groups. We observed no side effects for patients or damage to the instrument. CONCLUSIONS: The proposed technique could be a simple, safe, and inexpensive method for easier and less painful colonoscopy; moreover, it might facilitate difficult examinations. Further studies are needed to confirm our data and to ensure that the use of this technique is not liable to damage the scope. PMID- 15824945 TI - Significance of colonoscopy in patients with intestinal graft-versus-host disease after hematopoietic stem cell transplantation. AB - BACKGROUND AND STUDY AIMS: Patients with intestinal graft-versus-host disease (GVHD) are generally in poor clinical condition. In this study we aimed to establish the clinical significance of endoscopic diagnosis of this condition, observing only the distal section of the large intestine. PATIENTS AND METHODS: Endoscopic and pathological findings at colonoscopy were evaluated retrospectively in 12 patients who were diagnosed with intestinal GVHD after undergoing hematopoietic stem cell transplantation. RESULTS: The main mucosal changes observed at endoscopy were granular change, edema, "spotty redness", and sloughing. These were clearly displayed after enhancement with Indigo carmine staining, and with insertion of the colonoscope only as far as the distal section of the large intestine. A histological diagnosis of intestinal GVHD was made in 50 % of the patients, whose intestinal epithelium specimens showed numerous apoptotic bodies. It was possible to perform total colonoscopy in two patients who were in relatively good condition clinically, but there were no remarkable differences in the endoscopic findings throughout the large intestine, from the terminal ileum to the rectum. In terms of clinical outcomes of the 12 patients, their prognosis was poor in that they all either went on to suffer from chronic GVHD or died. CONCLUSIONS: Endoscopic and histological findings on distal colonoscopy are clinically significant in the diagnosis of intestinal GVHD, and limiting this examination to the distal section of the large intestine avoids causing further clinical deterioration in patients who are already in very poor general condition and the possibility of causing endoscopy-related complications. PMID- 15824946 TI - Clinical, colonoscopic, and histological profile of colonic tuberculosis in a tertiary hospital. AB - BACKGROUND AND STUDY AIMS: Colonic tuberculosis is not uncommon in developing countries. As emigration to the West increases, it is worthwhile to recall the clinical, colonoscopic, and histopathological features of this condition. PATIENTS AND METHODS: The clinical, colonoscopic and histopathological findings were evaluated in 43 patients with colonic tuberculosis. RESULTS: Abdominal pain, weight loss, diarrhea, fever, and a lump in the abdomen were the commonest symptoms. Extraintestinal tuberculosis was present in 11 patients (26 %). Colonoscopy revealed ulcers in 30 patients (70 %), nodules in 24 (56 %), a deformed cecum and ileocecal valve in 17 (40 %), strictures in 10 (23 %), polypoid lesions in six (14 %), and fibrous bands forming mucosal bridges in three (7 %). The cecum and ascending colon were the commonest sites involved. Segmental tuberculosis was seen in six of the 32 patients (19 %) in whom full length colonoscopy could be performed. Two or more sites were involved in 19 patients (44 %). Histopathology revealed well-formed granulomas in 23 patients (54 %). Fourteen of the above patients (61 %) had caseation and 11 (48 %) had confluence of the granulomas. Acid-fast bacilli were present in the biopsies from two patients (5 %). Ill-formed granulomas were seen in seven patients (16 %) and chronic inflammatory changes in 13 (30 %). Despite the various histopathological findings, all of the patients responded to antitubercular treatment and continued to remain asymptomatic during the follow-up period. CONCLUSIONS: Colonoscopy with biopsy is a useful method for diagnosing colonic tuberculosis. Even in the absence of the classic histopathological features, a therapeutic trial may be indicated in a given clinical and colonoscopic setting. Follow-up is essential. PMID- 15824947 TI - Value of endorectal ultrasonography for diagnosing rectovaginal septal endometriosis infiltrating the rectum. AB - BACKGROUND AND STUDY AIMS: Rectovaginal septal endometriosis (RVSE) can pose serious therapeutic problems when there is infiltration of the rectal septum (which occurs in approximately half of the cases). The aim of this study was to assess the value of endoscopic ultrasonography in diagnosing rectal wall involvement by pelvic endometriosis. PATIENTS AND METHODS: A prospective study was carried out from May 1998 to March 2003 at a single hospital center. The 30 patients included in the study presented with suspected RVSE and underwent systematic anorectal endoscopic ultrasonographic exploration prior to the surgical intervention. The endoscopic ultrasonography was carried out under general anesthesia with a 7.5-MHz miniprobe equipped with a distal balloon. RESULTS: The anorectal endoscopic ultrasonographic examination (EUS) showed the presence of endometriosis in the rectovaginal septum in 26 patients (88 %), in the uterosacral ligaments in 10 patients (33 %), and in the ovaries in two patients (6 %). At EUS, the nodules were infiltrating the rectal wall in 17 patients (56 %). The surgical exploration demonstrated endometriosis in the rectovaginal septum in 26 cases, the uterosacral ligaments in 22 cases, and the ovaries in 16 cases. The rectal wall was completely infiltrated in 12 cases and only partly in four cases, and intestinal tract resection was required in 10 cases. The sensitivity, specificity, and positive and negative predictive value of anorectal endoscopic ultrasonography as a means of diagnosing endometriosis of the rectovaginal septum and infiltration of the rectal wall were found to be 96 %, 100 %, 100 % and 83 %, and 92 %, 66 %, 64 % and 92 %, respectively; and the diagnostic accuracy was at 96 % and 80 %, respectively. The sensitivity for detecting nodules in the uterosacral ligaments or in the ovaries was 42 % and 14 %, respectively, leading to diagnostic accuracy rates of 56 % and 53 %. CONCLUSIONS: In terms of its sensitivity and its negative predictive value, anorectal endoscopic ultrasonography is a very effective means of detecting endometriosis of the rectovaginal septum and assessing possible infiltration of the rectal wall. However, this method is not as accurate for nodules located far from the EUS probe, as is the case with the uterosacral ligaments and ovaries. PMID- 15824948 TI - Puncture of solid pancreatic tumors guided by endoscopic ultrasonography: a pilot study series comparing Trucut and 19-gauge and 22-gauge aspiration needles. AB - BACKGROUND AND STUDY AIMS: The aim of this prospective study was to compare endoscopic ultrasonography-guided Trucut needle biopsy (EUS-TNB) with EUS-guided fine-needle aspiration biopsy (EUS-FNAB) using 19- and 22-gauge needles for biopsy from different sites in patients with solid pancreatic cancers. PATIENTS AND METHODS: Sixteen consecutive patients with masses in the uncinate process (n = 3), the head (n = 5), or the body and tail (n = 8) of the pancreas underwent both EUS-TNB and EUS-FNAB. The specimens obtained were evaluated by histopathological analysis alone RESULTS: Tissue specimens were obtained by Trucut needle, and by 19-gauge and 22-gauge aspiration needles in 69 %, 69 %, and 100 % of patients respectively. Sensitivity for malignancy was 69 % for all needles. Tissue sampling by Trucut and by 19-gauge aspiration needle from masses in the uncinate process was impossible. The sensitivity of the Trucut and 19 gauge aspiration needles was 100 % in the 11 patients with successful procedures. If Trucut or 19-gauge aspiration needles had been used for body and tail masses, and the 22-gauge aspiration needle for masses in the uncinate process and head, the sensitivity for malignancy would have been 81 %. CONCLUSIONS: EUS-TNB allows reliable tissue sampling for the diagnosis of pancreatic masses, but its use is limited to lesions in the body and tail of the pancreas. EUS-FNAB using a 22 gauge needle may be useful for accurate diagnosis in some patients with masses in the uncinate process or the head of the pancreas. PMID- 15824949 TI - UEGW Report, Prague 2004: endoscopic highlights. PMID- 15824950 TI - On writing (7): writing a book (2). PMID- 15824951 TI - Endoscopic ultrasound-guided fine-needle aspiration biopsy in esophageal cancer. PMID- 15824952 TI - Adaptive index of hemoglobin color enhancement for the diagnosis of colorectal disease. PMID- 15824953 TI - Therapeutic endoscopic retrograde cholangiopancreatography without fluoroscopy in four critically ill patients using wire-guided intraductal ultrasound. AB - Emergent endoscopic retrograde cholangiopancreatography cannot be performed at the bedside in critically ill patients in an intensive care unit because of the requirement for fluoroscopy. Moving such patients to a safe location where fluoroscopy is available can pose practical problems, and can lead to delayed intervention, which may adversely affect the outcome. We report the use of intraductal ultrasound to facilitate therapeutic biliary interventions in four critically ill patients in an intensive care unit. Cannulation was performed endoscopically at the bedside using a sphincterotome and a guide wire. Intraductal ultrasound, rather than fluoroscopy, was then used to confirm the location of the wire within the common bile duct prior to performing endoscopic sphincterotomy or stent placement. This technique was successful in all four patients. PMID- 15824954 TI - Drainage of gallbladder fossa fluid collections with endoprosthesis placement under endoscopic ultrasound guidance: a preliminary report of two cases. AB - Collections of fluid in the gallbladder fossa can be detected by ultrasound in as many as 29% of patients following cholecystectomy. Traditionally, persistent collections are treated by percutaneous drainage and bile duct decompression. We present two cases of persistent gallbladder fossa fluid collections which were refractory to bile duct decompression but which were successfully drained by endoscopic ultrasound-guided endoprosthesis placement. Under endoscopic ultrasound (EUS) control, a 19-gauge needle was inserted through the duodenal wall into the gallbladder fossa fluid collection. A guide wire was coiled within the collection, and an endoprosthesis was placed over the wire. Endoprosthesis insertion was successful in both cases, resulting in rapid symptomatic and radiographic improvement. EUS-guided drainage offers a minimally invasive alternative to percutaneous treatment of persistent gallbladder fossa fluid collections following cholecystectomy. PMID- 15824955 TI - Failure of capsule transportation due to gastroparesis after gastric truncal vagotomy. PMID- 15824956 TI - Use of the curved linear-array echo endoscope to identify gastrorenal shunts in patients with gastric fundal varices. PMID- 15824958 TI - Esophageal insertion of Polyflex stents without fluoroscopy in peptic strictures. PMID- 15824960 TI - Endoscopic identification of a gastric Dieulafoy's lesion following ephedrine administration. PMID- 15824961 TI - Endoclip-assisted closure of infected gastrocutaneous fistulas: report of two cases. PMID- 15824962 TI - Successful prevention of stent migration caused by placement of a second stent. PMID- 15824963 TI - Endoscopic resection of a large pedunculated colonic polyp using an insulated-tip diathermy knife. PMID- 15824964 TI - Successful endoscopic retrieval of an impacted trapezoid basket with severed traction wires. PMID- 15824965 TI - Diminutive gastrointestinal lesions in cholesterol crystal embolization. PMID- 15824966 TI - Zinc fluxes during acute and chronic exposure of INS-1E cells to increasing glucose levels. AB - Zinc in beta-cell secretory vesicles is essential for insulin hexamerization, and tight vesicular zinc regulation is mandatory. Little is known about zinc ion fluxes across the secretory vesicle membrane and the influence of changes in the extracellular environment on vesicular zinc. Our study aim was to investigate the effect of acute and chronic exposure to various glucose concentrations on zinc in secretory vesicles, the relation between zinc and insulin, and the presence of two zinc transporters, ZnT1 and ZnT4, in INS-1E cells. Zinc ions were demonstrated and semi-quantified using zinc-sulfide autometallography. Insulin content and secreted insulin were measured. Measurements were made on INS-1E cells after exposure to 2.0, 6.6, 16.7, and 24.6 mmol/l glucose for 1, 24, and 96 hours. 1h: Increasing glucose resulted in no changes in intravesicular zinc ions at 2, and 24.6 mmol/l glucose, but a slight increase at 16.7 mmol/l glucose. 24 and 96 h: Increasing glucose led to decreased vesicular zinc ion content accompanied by a decrease in insulin content. ZnT1 and ZnT4 were present in the cytoplasm. Our results demonstrate that intra-vesicular zinc ions respond to changes in the extra-cellolar glucose concentration, especially during chronic high glucose concentrations, where the content of vesicular zinc ions decreases. PMID- 15824967 TI - SAPK/JNK plays a role in transforming growth factor-beta-induced VEGF synthesis in osteoblasts. AB - We previously reported that transforming growth factor-beta (TGF-beta) activates p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase, resulting in the stimulation of vascular endothelial growth factor (VEGF) synthesis in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the involvement of stress-activated protein kinase/c- Jun N-terminal kinase (SAPK/JNK), another member of the MAP kinase superfamily, in TGF-beta-induced VEGF synthesis in these cells. TGF-beta markedly induced SAPK/JNK phosphorylation. SP600125, a specific inhibitor of SAPK/JNK, markedly reduced TGF beta-induced VEGF synthesis. SP600125 suppressed TGF-beta-induced SAPK/JNK phosphorylation. PD98059, an inhibitor of upstream kinase of p44/p42 MAP kinase and SB203580, an inhibitor of p38 MAP kinase, each failed to reduce TGF-beta induced SAPK/JNK phosphorylation. A combination of SP600125 and PD98059 or SP600125 and SB203580 suppressed TGF-beta-stimulated VEGF synthesis in an additive manner. These results strongly suggest that TGF-beta activates SAPK/JNK in osteoblasts, and that SAPK/JNK plays a role in addition to p42/p44 MAP kinase and p38 MAP kinase in TGF-beta-induced VEGF synthesis. PMID- 15824968 TI - Improvement of insulin resistance by panax ginseng in fructose-rich chow-fed rats. AB - In an attempt to probe a new target for handling insulin resistance, we used Panax ginseng root to screen the effect on insulin resistance induced by fructose rich chow in rats. Insulin action on glucose disposal rate was measured using the glucose-insulin index, which is the product of the areas under the curve of glucose and insulin during the intraperitoneal glucose tolerance test. Oral administration of Panax ginseng root (125.0 mg/kg) into rats three times daily for three days after receiving fructose-rich chow for four weeks reversed the increased glucose-insulin index, indicating that Panax ginseng root has the ability to improve insulin sensitivity. In addition, the plasma glucose concentrations in rats repeatedly treated with Panax ginseng root were not elevated as markedly as those of the vehicle-treated group during the fructose rich chow-feeding period. Also, the time in which the plasma glucose-lowering response to tolbutamide (10.0 mg/kg, i. p.) receded in fructose-rich chow fed rats was markedly delayed by repeated Panax ginseng root treatment compared to the vehicle-treated group. The plasma glucose-lowering activity of tolbutamide is believed to depend on the secretion of endogenous insulin, which is widely used as an indicator of insulin resistance development. Thus, it provided supportive data that oral administration of Panax ginseng root could delay the development of insulin resistance in rats. In conclusion, our results suggest that oral administration of Panax ginseng root improves insulin sensitivity and may be used as an adjuvant therapy for treating diabetic patients with insulin resistance. PMID- 15824969 TI - Leptin concentrations in plasma and follicular fluid from prepubertal gilts as influenced by fasting, refeeding and insulin. AB - This study's aim was to examine whether fasting and refeeding would influence leptin levels in both plasma and follicular fluid from prepubertal gilts, and whether insulin affects leptin levels in fasting gilts. In experiment 1, four gilts were fasted for 72 h and then refed. Blood samples were withdrawn during normoalimentation, at the end of fasting, and for 4 h after refeeding. All samples were assayed for leptin; alternate samples were assayed for insulin, glucose and non-esterified fatty acids (NEFA). Fasting caused a decrease in leptin, glucose and insulin levels in plasma, while NEFA concentrations increased. In experiment 2, four gilts were given insulin as a bolus (0.2 IU/kg body weight) after 68 h of fasting. Blood samples were collected every 15 min around insulin administration and were assayed for leptin, insulin and glucose. This experiment shows that insulin administration increases leptin levels during fasting. In experiment 3, gilts were ovariectomized during normal alimentation (n=4), after 48 h of fasting (n=4), and after 48 h of realimentation following 48 h of fasting (n=4). Leptin levels in both plasma and follicular fluid collected after 48 h of fasting were significantly lower than those observed during normoalimentation or refeeding. In conclusion, a transient increase in insulin during fasting is effective in restoring leptin concentrations; in addition, leptin levels in follicular fluid parallel those in plasma. PMID- 15824970 TI - PNMT transgenic mice have an aggressive phenotype. AB - PNMT (phenylethanolamine-N-methyl-transferase) is the enzyme that catalyzes the formation of epinephrine from norepinephrine. In transgenic mice over-expressing PNMT, observations revealed a very high level of aggression compared to their background strain, C57BL/6J. To evaluate the influence of PNMT on aggression and emotionality in this transgenic line, single-sex male and female groups were independently established that consisted of either four wild-type mice or four transgenic mice overexpressing PNMT. The members of each group were littermates. Mixed single-sex groups consisting of two transgenic mice and two wild-type mice were also established. Almost no fights were observed within the female groups. In males, the transgenic line showed a significantly higher level of fighting than controls (p=0.007) and mixed male groups (p=0.02). Housing mice from the transgenic line in mixed groups with wild-type mice seems to decrease the level of aggression in the transgenic line. In conclusion, this is the first study to demonstrate a clear, significant increase in aggression arising from PNMT overexpression. This suggests an important role for central epinephrine levels in aggressive behavior. PMID- 15824971 TI - Troglitazone reduces leptinemia during experimental dexamethasone-induced stress. AB - BACKGROUND AND AIMS: Recently, we found that profound anorexia observed in a catabolic model induced by chronic glucocorticoid (dexamethasone, Dex) injection could be associated with strong hyperleptinemia. To investigate the implication of leptin in this catabolic stress response, we used a model whereby leptin secretion was inhibited using troglitazone (Trg) concomitantly with a Dex-induced stress injection. METHODS: Adult rats (3 months, n=12) were stressed with a Dex injection (1.5 mg/kg/day ip, 5 days) and either treated (DXTG+, n=6) or not (DXTG , n=6) with Trg (60 mg/kg/day sc, 5 days). These DXTG+ and DXTG- groups were compared with an untreated ad libitum group and a pair-fed group receiving saline ip instead of the Dex injection. The effects of troglitazone treatment on leptin gene expression in adipose tissue, blood glucose, insulin, and on hepatic parameters under stress conditions were determined. RESULTS: Trg treatment specifically diminished leptinemia (30%, DXTG+ vs DXTG-, p<0.05). Insulinemia and glycemia remained unchanged, as did leptin gene expression; food intake improved, but hepatic capacities did not show any alteration. CONCLUSION: Trg is a useful agent in exploring certain potential effects of leptin on metabolic and immune disorders occurring during aggression. PMID- 15824972 TI - Three-generation investigation on serotonin content in rat immune cells long after beta-endorphin exposure in late pregnancy. AB - Female rats were treated with beta-endorphin on the 19th day of pregnancy. Serotonin content of immune cells (peritoneal lymphocytes, monocyte-macrophage granulocyte group (mo-gran), mast cells, blood lymphocytes, granulocytes and monocytes, thymus lymphocytes) were studied in the mothers (P-generation four weeks after delivery), in the male offspring (F1) generation (at seven weeks), in the female offspring (four weeks after their own delivery) and in their offspring (F2 generation, at seven weeks). P-mother cells' serotonin content was not influenced by endorphin treatment, while F1 generation's mo-gran and blood lymphocyte serotonin content was reduced (in contrast, histamine content of mo gran increased). Four weeks after delivery, an increase in serotonin content was observed in the F1 generation in the peritoneal lymphocytes and mast cells as well as in blood lymphocytes. In contrast, serotonin content was reduced in blood granulocytes and monocytes. In the F2 (grandson) generation, a reduction in mast cell serotonin content and sensitization of blood and thymic lymphocytes to repeated endorphin treatment was provoked. The significant changes were more expressed in the F2 generation compared to F1, also appearing earlier. The results unequivocally suggest that the increase in endorphin levels during late pregnancy can cause permanent changes in the F1 and F2 generations, which means that the imprinting effect can be transgenerationally transmitted. PMID- 15824973 TI - Effects of estrogen replacement therapy on bone and glucose metabolism in a male with congenital aromatase deficiency. AB - Little is known about the impact of estrogen replacement therapy for bone formation, glucose metabolism and hormonal parameters on males with aromatase deficiency. Transdermal estrogen (TE) replacement was initiated at 100 microg/week in months 0-3, 50 microg/week in months 3-6, 25 microg/week in months 6-12, 75 microg/week in months 12-24, and 25 microg/week in months 24-36 to substitute for the deficiency in a 27-year-old homozygous male with a mutation on the CYP19 gene. Estradiol levels increased from<10 at baseline to 45, 12, 27 and 17 pg/ml (normal range 10-50) after 6, 12, 24 and 36 months, and inversely correlated to LH and FSH levels. Testosterone levels changed from 31.2 nmol/l at baseline to 3.8, 22.1, 7.1 and 22.0 nmol/l (9.5-30) after 6, 12, 24 and 36 months, respectively, and correlated closely to basal and stimulated LH and FSH levels at 100 microg GnRH. Bone maturation progressed, and metacarpal and phalangeal epiphysis closed after 12 months. Spongiosa-hydroxyapatite of the radius assessed by quantitative computed tomography changed from 52 to 83, 51, 69 and 71 mg/cm3 (120-160); bone mineral density of the lumbar spine assessed by dual energy X-ray-absorptiometry (normal value>1.150) increased from 0.971 (T Score -2.24) to 1.043 (-1.64), 1.065 (-1.46), 1.128 (-0.93) g/cm2 and 1.021 ( 1.82) after 6, 12, 24 and 36 months of TE, respectively. Osteocalcin as a bone formation parameter and aminoterminal collagen type I telopeptide as a bone resorption parameter increased during high-dose estrogen supplementation, and then decreased during the lower doses. Lipoprotein (a) increased from 20 mg/dl at baseline to 60 and 62 mg/dl after 6 and 12 months, and then decreased to 24 and 25 mg/dl after 24 and 36 months, respectively, while total cholesterol, HDL, LDL and triglycerides did not change. AUC glucose decreased continuously after oral glucose load, and HOMA IR reached its lowest value the 75 microg weekly estradiol dose. This study confirms the role of estrogens in achieving bone mineralization and maturation in human males. Additionally, estradiol has dual negative feedback sites that on the hypothalamus to decrease GnRH pulse frequency, and on the pituitary to decrease responsiveness to GnRH. The improvement in glucose metabolism after estrogen replacement therapy suggests a probable role of sex steroids in insulin sensitivity. The optimal weekly dose of transdermal estrogen replacement for normalizing estrogen levels and maintain bone mass in adult males with aromatase deficiency may be 50-75 microg spread over two doses. PMID- 15824974 TI - Sex hormones in patients with epilepsy-hormonal changes in epileptic men and women taking antiepileptics. AB - The relationship between epilepsy and endocrine system has attracted the attention of investigators for a number of years. Epilepsy is a common neurological disorder; both seizures and antiepileptic drugs can compromise the physical and hormonal aspects of sexual development. Impairment of libido and sexual potency have been frequently reported in male epileptic patients. Women with epilepsy have a greater risk of infertility (anovulatory cycles and polycystic ovary syndrome). This review analyses the main data from the literature in order to clarify the role of epilepsy and antiepileptic drugs on sex hormones in epileptic patients. As gonad dysfunction is frequently observed in women and men with epilepsy, particularly when taking antiepileptic drugs, ovarian and testicular function must be carefully monitored. PMID- 15824975 TI - The effect of short-term cold and hot exposure on total plasma ghrelin concentrations in humans. PMID- 15824976 TI - Focal neurological deficits and West Nile virus infection. AB - Our experience with West Nile virus infection revealed that 54% of 28 patients had a focal neurological deficit at presentation. A meningitis or encephalitis syndrome was absent in 47% of patients with focal deficits. Details of the variety of deficits, the time to development of deficits, and the associated radiological and laboratory findings are also discussed in the present report. PMID- 15824977 TI - Milkers' nodules complicated by erythema multiforme and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation for multiple myeloma. AB - We describe a case of cow-transmitted parapoxvirus infection--also known as milkers' nodules--after a hematopoietic stem cell transplantation for multiple myeloma. The infection was complicated by erythema multiforme and acute exacerbation of graft-versus-host disease. Parapoxvirus was confirmed by electron microscopy. The natural history of milker's nodules in immunocompetent hosts is described and compared to that in our immunocompromised patient. PMID- 15824978 TI - Anaerobic sepsis due to multidrug-resistant Bacteroides fragilis: microbiological cure and clinical response with linezolid therapy. AB - We describe the first reported case of anaerobic sepsis due to Bacteroides fragilis with simultaneous resistance to metronidazole, beta-lactams, beta lactam/beta-lactamase inhibitors, carbapenems, macrolides, and tetracyclines. Microbiological cure and clinical improvement was achieved with linezolid therapy, an agent that may be useful for the treatment of multidrug-resistant anaerobic infections. PMID- 15824979 TI - Changing bacteriology of adult community-acquired lung abscess in Taiwan: Klebsiella pneumoniae versus anaerobes. AB - BACKGROUND: Most literature regarding lung abscess focuses on anaerobic bacterial lung abscess, and aerobic gram-negative bacillary infection is less frequently discussed. This study was conducted to investigate the bacteriology of community acquired lung abscess and to improve the empirical therapeutic strategy for adults with community-acquired lung abscess. METHODS: We reviewed and analyzed data on 90 consecutive adult cases of bacteriologically confirmed community acquired lung abscess treated during 1995-2003 at a tertiary university hospital in Taiwan. RESULTS: We found that a high proportion (21%) of cases of lung abscess were due to Klebsiella pneumoniae infection, which differs from the findings of previous studies. Lung abscess due to K. pneumoniae was associated with underlying diabetes mellitus (odds ratio [OR], 4.3; 95% confidence interval [CI], 1.0-18.4; P = .039) and negatively correlated with a time from onset of symptoms to diagnosis of >30 days (OR, 0.2; 95% CI, 0.1-0.7; P = .008). A higher percentage of patients with K. pneumoniae lung abscess had concomitant bacteremia (OR, 9.4; 95% CI, 1.1-81.9; P = .032), delayed defervesence (OR, 9.2; 95% CI, 1.8 47.8; P = .004), and multiple cavities noted on radiographs (OR, 11.0; 95% CI, 1.3-94.9; P = .015), compared with patients with anaerobic bacterial lung abscess. The rate of nonsusceptibility to clindamycin and penicillin among anaerobes and Streptococcus milleri group isolates increased. CONCLUSION: K. pneumoniae has become a more common cause of lung abscess than before, and a high proportion of anaerobes and S. milleri strains have become resistant to penicillin and clindamycin. A beta-lactam/beta-lactamase inhibitor or second- or third-generation cephalosporin with clindamycin or metronidazole is suggested as empirical antibiotic therapy for community-acquired lung abscess. PMID- 15824980 TI - The role of anaerobic bacteria in lung abscess. PMID- 15824981 TI - Surveillance of nosocomial sepsis and pneumonia in patients with a bone marrow or peripheral blood stem cell transplant: a multicenter project. AB - BACKGROUND: For surveillance of nosocomial bloodstream infections (BSIs) and pneumonia during neutropenia in adult patients who have undergone bone marrow transplantation (BMT) or peripheral blood stem cell transplantation (PBSCT), a multicenter study--the Hospital Infection Surveillance System for Patients with Hematologic/Oncologic Malignancies (ONKO-KISS)--was initiated in Germany in 2000. METHODS: Nosocomial infections were identified in neutropenic patients by means of Centers for Disease Control and Prevention definitions for laboratory confirmed BSI and modified criteria for pneumonia. RESULTS: During the first 38 month period of the study (i.e., through December 2003), a total of 1899 patients associated with 28,273 neutropenic days were investigated. Of these, 1173 (62%) had undergone allogeneic and 726 (38%) had undergone autologous BMT or PBSCT. The mean duration of neutropenia was 14.9 days (9.6 and 18.1 days after autologous and allogeneic transplantation, respectively). Overall, 395 BSIs and 168 cases of pneumonia were identified. The pooled mean site-specific incidence density per 1000 neutropenic days was 14.0 for BSI (12.4 and 18.9 for the allogeneic and autologous transplantation groups, respectively) and 5.9 for pneumonia (6.1 and 5.6 in the allogeneic and autologous transplantation groups, respectively). After allogeneic transplantation, 22.4 BSIs per 100 patients and 11.0 cases of pneumonia per 100 patients occurred, whereas 18.2 BSIs per 100 patients and 5.4 cases of pneumonia per 100 patients occurred after autologous transplantation. The majority (57%) of pathogens associated with BSI were coagulase-negative staphylococci. CONCLUSIONS: The ongoing ONKO-KISS project provides unprecedented reference data about the incidence of pneumonia and sepsis among BMT recipients and PBSCT recipients in Germany. These data will be used for further evaluation of the impact of hygiene measures and therapeutic regimens for these patients. PMID- 15824982 TI - Clinical outcomes of human herpesvirus 6 reactivation after hematopoietic stem cell transplantation. AB - BACKGROUND: Although human herpesvirus 6 (HHV-6) is known to reactivate during hematopoietic stem cell transplantation (HSCT), the clinical significance of this finding is controversial. METHODS: We used a quantitative PCR test for HHV-6 to assay plasma samples prospectively collected from a cohort of 110 allogeneic HSCT recipients to evaluate the clinical effects of HHV-6 infection. A retrospective review of medical records was performed to determine clinical end points. RESULTS: HHV-6 reactivation occurred in 52 (47%) of the 110 subjects. Factors that increased the risk of subsequent HHV-6 reactivation were hematologic malignancy that occurred at a time other than the first remission (adjusted P = .002), a mismatch in the sexes of donor and recipient (adjusted P=.05), younger age (adjusted P = .01), and the receipt of glucocorticoids (adjusted P = .06). HHV-6 reactivation was associated with subsequent all-cause mortality (adjusted hazard ration [HR], 2.9; 95% confidence interval [CI], 1.1-7.5), grade 3-4 graft versus-host disease (GVHD) (adjusted HR, 4.9; 95% CI, 1.5-16), a lower probability of monocyte engraftment (adjusted HR, 0.42; 95% CI; 0.22-0.80), a lower probability of platelet engraftment (adjusted HR, 0.47; 95% CI, 0.21-1.1; P = .05) and a higher platelet transfusion requirement (adjusted P = .02). A higher level of HHV-6 DNA was associated with subsequent central nervous system (CNS) dysfunction (HR, 21; 95% CI, 1.8-249). CONCLUSIONS: HHV-6 reactivation is common after allogeneic HSCT and is associated with subsequent delayed monocyte and platelet engraftment, increased platelet transfusion requirements, all-cause mortality, grade 3-4 GVHD, and CNS dysfunction. PMID- 15824983 TI - Purpura fulminans due to Staphylococcus aureus. AB - BACKGROUND: Purpura fulminans is an acute illness commonly associated with meningococcemia or invasive streptococcal disease, and it is typically characterized by disseminated intravascular coagulation (DIC) and purpuric skin lesions. In this article, we report the first 5 cases (to our knowledge) of purpura fulminans directly associated with Staphylococcus aureus strains that produce high levels of the superantigens toxic shock syndrome toxin-1 (TSST-1), staphylococcal enterotoxin serotype B (SEB), or staphylococcal enterotoxin serotype C (SEC). METHODS: Cases were identified in the Minneapolis-St. Paul, Minnesota, metropolitan area during 2000-2004. S. aureus infection was diagnosed on the basis of culture results, and susceptibility to methicillin was determined. The ability of the isolated organisms to produce TSST-1, SEB, SEC, and Panton-Valentine leukocidin (PVL) was determined. TSST-1, SEB, and SEC levels were also quantified after in vitro growth of the organisms. RESULTS: In 3 of the 5 cases, the infecting S. aureus strain was isolated from the blood cultures. In 2 of the 5 cases, the infecting S. aureus strain was isolated only from the respiratory tract, indicating that purpura fulminans and toxic shock syndrome resulted from exotoxin and/or other host factors, rather than septicemia. One of these latter 2 patients also had necrotizing pneumonia, and the isolated S. aureus was a methicillin-resistant strain that produced both SEC and PVL. Only 2 of the 5 patients survived, and 1 of the survivors received activated protein C. CONCLUSIONS: Staphylococcal purpura fulminans may be a newly emerging illness associated with superantigen production. Medical practitioners should be aware of this illness. PMID- 15824984 TI - Staphylococcal purpura fulminans: a toxin-mediated disease? PMID- 15824985 TI - Prospective evaluation of community-acquired acute-phase hepatitis C virus infection. AB - BACKGROUND: More than two-thirds of hepatitis C virus (HCV) infections in Western countries are caused by injection drug use, but prospective clinical data regarding the most common mode of HCV acquisition are rare, in part because acute phase HCV infection is usually asymptomatic. METHODS: To characterize acute-phase HCV infection, 179 HCV antibody-negative injection drug users were prospectively evaluated; 62 (34%) of these patients had seroconverted. Twenty of the participants who seroconverted had long-term follow-up with consistent monthly sampling before and after seroconversion, allowing detailed study. RESULTS: The first indication of HCV infection was the presence of HCV RNA in serum, which preceded elevation of alanine transaminase levels and total bilirubin levels to > or =2 times baseline in 45% and 77% of patients, respectively. No subjects had jaundice. The median time from initial viremia to seroconversion was 36 days (range, 32-46 days). In one instance, viremia was detected 434 days before seroconversion. However, in no other case was HCV RNA detected >63 days before seroconversion. In subjects with viral persistence, a stable level of HCV RNA in the blood was noted in some subjects within 60 days after the initial detection of viremia, but in others, it was not apparent until >1 year later. In subjects with long-term viral clearance, HCV became persistently undetectable as early as 94 and as late as 620 days after initial viremia. CONCLUSIONS: These data underscore the importance of nucleic acid screening of blood donations to prevent HCV transmission and of long-term follow-up to ascertain whether there is viral persistence, at least among injection drug users. PMID- 15824986 TI - Acute-phase hepatitis C virus infection: implications for research, diagnosis, and treatment. PMID- 15824987 TI - Multistate outbreak of Listeria monocytogenes infection linked to delicatessen turkey meat. AB - BACKGROUND: Despite a decreasing incidence of listeriosis in the United States, molecular subtyping has increased the number of recognized outbreaks. In September 2000, the New York City Department of Health identified a cluster of infections caused by Listeria monocytogenes isolates with identical molecular subtypes by pulsed-field gel electrophoresis (PFGE) and ribotyping. METHODS: To determine the magnitude of the outbreak and identify risk factors for infection, we notified state health departments and conducted a case-control study. A case was defined as a patient or mother-infant pair infected with Listeria monocytogenes whose isolate yielded the outbreak PFGE pattern. Controls were patients infected with Listeria monocytogenes whose isolate yielded a different PFGE pattern. Patients were asked about food and drink consumed during the 30 days before the onset of illness. RESULTS: Between May and December 2000, there were 30 clinical isolates of Listeria monocytogenes with identical PFGE patterns identified in 11 US states. Cases of infection caused by these isolates were associated with 4 deaths and 3 miscarriages. A case-control study implicated sliced processed turkey from a delicatessen (Mantel-Haenszel odds ratio, 8.0; 95% confidence interval, 1.2-43.3). A traceback investigation identified a single processing plant as the likely source of the outbreak, and the company voluntarily recalled 16 million pounds of processed meat. The same plant had been identified in a Listeria contamination event that had occurred more than a decade previously. CONCLUSIONS: Prevention of persistent L. monocytogenes contamination in food processing plants presents a critical challenge to food safety professionals. PMID- 15824988 TI - Treatment of multidrug-resistant tuberculosis in San Francisco: an outpatient based approach. AB - BACKGROUND: Treatment of patients with multidrug-resistant tuberculosis requires prolonged therapy, often involving long hospital stays. Despite intensive and costly therapy, cure rates are relatively low. METHODS: We reviewed the outcomes for all patients with multidrug-resistant tuberculosis treated in San Francisco, California, during 1982-2000 and identified billing charges for patients treated during 1995-2000. Mycobacterium tuberculosis isolates were genotyped by IS6110 based restriction fragment-length polymorphism analysis. RESULTS: Forty-eight cases were identified with resistance to a median of 3 drugs (range, 2-9 drugs). The median age of the patients was 49.5 years (range, 22-78 years); 36 (75%) of 48 patients were foreign born, 11 (23%) were human immunodeficiency virus (HIV) seropositive, and 45 (94%) had pulmonary tuberculosis. Thirty-two (97%) of the 33 HIV-seronegative patients were cured, with only 1 relapse occurring 5 years after treatment. All 11 HIV-seropositive patients died during observation. Twenty-one patients (44%) required hospitalization, with a median duration of stay of 14 days (range, 3-74 days). The estimated inpatient and outpatient aggregate cost for the 11 patients treated after 1994 was $519,928, with a median cost of $27,752 per patient. No secondary cases of multidrug-resistant tuberculosis were identified through population-based genotyping. CONCLUSIONS: Treatment of multidrug-resistant tuberculosis in HIV-seronegative patients largely on an outpatient basis was feasible and was associated with high cure rates and lower cost than in other published studies. Patients with underlying HIV infection had very poor outcomes. PMID- 15824989 TI - Identification of methicillin-resistant Staphylococcus aureus carriage in less than 1 hour during a hospital surveillance program. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has spread worldwide and is responsible for significant morbidity, mortality, and health care costs. Control strategies to limit the emergence and spread of this organism rely on rapid and sensitive tests for detection of MRSA carriage. However, the standard surveillance culture method for detecting MRSA is labor intensive and time-consuming (2-3 days per procedure). There is thus a need for a rapid and accurate method to screen for MRSA carriage. METHODS: We recently developed an easy-to-use real-time polymerase chain reaction (PCR) assay suitable for specific detection of MRSA in nasal specimens in <1 h. We studied the efficacy of our new PCR assay in routine screening for nasal MRSA carriage during a hospital surveillance program. A total of 331 nasal specimens obtained from 162 patients at risk for colonization were tested by both the standard mannitol agar culture method and our PCR assay. RESULTS: The PCR assay detected MRSA in all 81 samples that were culture positive for MRSA. The PCR assay detected 4 additional MRSA positive specimens, for a specificity of 98.4%, a positive predictive value of 95.3%, and a sensitivity and negative predictive value of 100%. CONCLUSIONS: This novel PCR assay allows reliable identification of MRSA carriers in <1 h. This test should facilitate the efficacy of MRSA surveillance programs. PMID- 15824990 TI - Diagnosis of enteroviral meningitis by use of polymerase chain reaction of cerebrospinal fluid, stool, and serum specimens. AB - BACKGROUND: Because enteroviruses can be detected in various clinical samples during enteroviral meningitis, we analyzed the combined diagnostic utility of polymerase chain reaction (PCR) of cerebrospinal fluid (CSF), feces, and serum for detection of enterovirus in specimens obtained from adults with aseptic meningitis or encephalitis. METHODS: PCR results were analyzed for 34 adults for whom enteroviral meningitis was diagnosed on the basis of virus isolation and antibody detection in our hospital during 1999-2003. PCR results were also analyzed for 77 adults with meningitis or encephalitis of another defined cause for whom this assay was used for diagnostic evaluation during that period. RESULTS: Twenty-six (76%) of 34 CSF samples and 24 (96%) of 25 fecal samples collected from patients with enteroviral meningitis had positive PCR results. The diagnostic yield of the test was lower for CSF specimens obtained >2 days after clinical onset, compared with CSF collected < or =2 days after onset. Instead, PCR of feces was highly useful also later, because 12 of the 13 fecal specimens obtained 5-16 days after clinical onset had positive test results. None of 75 CSF samples and 2 of 48 fecal samples obtained from patients with nonenteroviral infection had positive PCR results. All serum samples were PCR negative. CONCLUSIONS: PCR of fecal specimens obtained throughout the course of enteroviral meningitis had the highest clinical sensitivity for detecting enterovirus. It is recommended that, in addition to performance of CSF PCR, fecal samples collected from patients with suspected enteroviral meningitis should be tested by PCR, especially when the duration of symptoms is >2 days. PMID- 15824991 TI - Serious infections in elderly patients with diabetes mellitus. AB - The elevated serum glucose levels associated with diabetes mellitus (DM) alter host immune responses, resulting in a well-documented increase in the predisposition to infectious processes. Furthermore, the cumulative effect of age related immune senescence, superimposed on this enhanced risk of infections, can lead to serious and life-threatening infections in elderly patients with DM. Because infection associated with aging can frequently present in a subtle and atypical manner, prompt recognition of infection and treatment with appropriate empirical broad-spectrum antimicrobial agents, in conjunction with surgical intervention, is often necessary to eradicate such infections. Common sites of serious infection associated with DM include the head and neck, biliary tract, and urinary tract, as well as the skin, soft tissue, and bony structures of the feet in particular. PMID- 15824992 TI - Unique aspects of antimicrobial use in older adults. AB - Elderly persons consume a significant proportion of health care resources. Antimicrobials are just one class among many pharmaceuticals that are prescribed more frequently to elderly patients than to younger patients. There are unique aspects of antimicrobial use in elderly persons that make prescribing complicated and monitoring unpredictable. Physiologic changes associated with aging result in altered pharmacokinetics, and accurate estimates of renal function cannot be made with standard methods. Together, these qualities make antimicrobial dosing difficult. Because of a higher prevalence of other chronic diseases, there is a greater propensity for polypharmacy and a resulting risk of an adverse event or a significant drug interaction. Lastly, irrespective of altered pharmacokinetics, adverse effects of many antimicrobials are more common in elderly persons, which introduces an added dimension to ensuring safety with antimicrobial therapy. PMID- 15824994 TI - Invasive infection with multidrug-resistant Salmonella enterica serotype typhimurium definitive type 104 among HIV-infected adults. AB - BACKGROUND: Multidrug-resistant Salmonella enterica serotype Typhimurium definitive type 104 (MRDT104), with resistance to at least ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (R-type ACSSuT), was first detected in the United States in 1985, and the prevalence increased to account for nearly 7% of Salmonella infections in 1998. METHODS: A retrospective study of S. Typhimurium infections in an urban health care system assessed whether infection with an antibiotic-resistant strain--and specifically MRDT104--was associated with invasive disease or HIV infection. Sixty cases of S. Typhimurium infection were identified. RESULTS: Of the 50 isolates available for analysis, 30 (60%) were MRDT104. Pathogens were isolated from blood in 25 (83%) of 30 patients infected with MRDT104, compared with 10 (50%) of 20 patients who were infected with non-MRDT104 strains (P = .01). Among isolates obtained from 32 HIV-infected patients, 19 (95%) of 20 MRDT104 isolates were from blood specimens, compared with 8 (66%) of 12 non-MRDT104 isolates (P = .05). CONCLUSIONS: MRDT104 accounted for the majority of S. Typhimurium infections in this patient population, and MRDT104 infections were more invasive than non-MRDT104 infections, particularly in HIV-infected persons. PMID- 15824995 TI - Characterization of immunologic and virological parameters in HIV-infected patients with primary effusion lymphoma during antiblastic therapy and highly active antiretroviral therapy. AB - BACKGROUND: Primary effusion lymphoma (PEL) represents a peculiar lymphoma infected with human herpesvirus 8 (HHV-8) and occurs predominantly in human immunodeficiency virus (HIV)-infected patients. The aim of the present study was to evaluate the immunologic and virological parameters, including HHV-8 viremia, of 5 HIV-infected patients with PEL whose disease was diagnosed and treated at our institute. METHODS: Five patients were enrolled in the study. Biological parameters, such as latent and lytic HHV-8 antigen levels, plasma HHV-8 load, Epstein-Barr virus plasma DNA load, HIV-1 load, and CD4 cell count, were assessed before treatment, during therapy, and at follow-up. RESULTS: Four patients were treated with chemotherapy and highly active antiretroviral therapy (HAART), and 1 was treated with HAART alone; 3 of 5 patients reached complete remission. HHV-8 could be detected before the initiation of therapy in plasma from all patients analyzed. HHV-8 levels decreased after therapy in 4 patients. During the whole observation period, plasma HHV-8 load showed a statistically significant inverse correlation with CD4 cell count but no significant correlation with HIV load and response to therapy. CONCLUSIONS: Our analysis demonstrates that HHV-8 can be detected in the plasma at the onset of PEL; its prognostic role needs to be explored. CD4 cell count seems to be the most important indicator of progression of PEL. PMID- 15824993 TI - Parasitic central nervous system infections in immunocompromised hosts. AB - Immunosuppression due to therapy after transplantation or associated with HIV infection increases susceptibility to various central nervous system (CNS) infections. This article discusses how immunosuppression modifies the presentation, diagnosis, and treatment of selected parasitic CNS infections, with a focus on toxoplasmosis, Chagas disease, neurocysticercosis, schistosomiasis, and strongyloidiasis. PMID- 15824996 TI - A molecular epidemiological analysis of 2 Staphylococcus aureus clonal types colonizing and infecting patients with AIDS. AB - BACKGROUND: Persons with acquired immune deficiency syndrome (AIDS) who use drugs appear to be at increased risk for colonization and infection with Staphylococcus aureus. Little is known about the nature of and risk factors responsible for this association. This study is among the first to prospectively follow carriage and infection in this uniquely high-risk population. METHODS: We prospectively followed the cases of 75 patients with AIDS in a residential drug treatment facility and screened for S. aureus nasal colonization and infection. RESULTS: Thirty-seven baseline cultures (49%) were positive for S. aureus, and 81% of subjects were colonized at least once during the study. Thirteen subjects experienced 17 infections. Pulsed-field gel electrophoresis and sequence-based typing methods revealed that 244 (92%) of the isolates belonged to either clonal type A or B. Clonal type A was methicillin-susceptible. Clonal type B consisted of 3 main subtypes (B1, B2, and B3), all with the same allelic profile (ST8) and staphylococcal protein A gene (spa) type (7). Of note, subtype B1 was methicillin susceptible (ST8 and spa type 7), lacking mecA, whereas the other B clones were methicillin-resistant. Both clones were resistant to trimethoprim sulfamethoxazole. Clonal type B isolates were relatively resistant, suggesting prior exposure to the health care setting. CONCLUSIONS: This study demonstrates a sustained high rate of S. aureus carriage and infection. It demonstrates the capacity of unique methicillin-resistant S. aureus clones with an established linkage to earlier outbreaks of methicillin-resistant S. aureus, as well as to human immunodeficiency virus--infected subjects, to persist in this residential setting. It also illustrates the apparent genetic instability or transmissibility of the staphylococcal chromosomal cassette mec type IV element. PMID- 15824997 TI - It is time to implement routine, not risk-based, HIV testing. AB - Approximately one-quarter of a million persons in the United States who are infected with human immunodeficiency virus (HIV) do not know it. To decrease the number of such persons, primary care providers should make HIV testing a routine component of health care. HIV testing should also be offered routinely in other settings, such as emergency departments, jails, and substance abuse treatment centers. Currently, the Centers for Disease Control and Prevention and the Infectious Diseases Society of America recommend routine HIV testing only in settings where the prevalence of HIV infection is > or =1%; in settings where the prevalence of HIV infection is <1%, testing should be based on risk assessment. Because of the impracticality of strategies for testing that are based on estimates of prevalence, and because of the inaccuracy of risk assessment, we propose that HIV testing be routinely offered to any person who is sexually active. As an adjunct to the implementation of routine testing programs, counseling practices need to be streamlined, and rapid HIV testing needs to be implemented in the appropriate settings. PMID- 15824998 TI - Establishment of a public antiretroviral treatment clinic for adults in urban Botswana: lessons learned. AB - Countries in sub-Saharan Africa are under significant pressure to open large scale, public antiretroviral treatment clinics. Many lessons have been learned in Botswana, where the first public antiretroviral treatment clinic in Africa was established. The availability of core, well-trained medical staff will be the primary factor that limits a rapid scale-up of antiretroviral treatment programs. PMID- 15824999 TI - HIV/AIDS care in Africa today. PMID- 15825000 TI - The role of immune reconstitution inflammatory syndrome in AIDS-related Cryptococcus neoformans disease in the era of highly active antiretroviral therapy. AB - This study of human immunodeficiency virus (HIV)-infected patients coinfected with Cryptococcus neoformans found that 30% of patients who initiated highly active antiretroviral therapy developed immune reconstitution inflammatory syndrome (IRIS). Patients with C. neoformans-related IRIS had higher cerebrospinal fluid opening pressures, glucose levels, and white blood cell counts, compared with patients with typical HIV-associated C. neoformans meningitis. PMID- 15825001 TI - Varicella-associated invasive group A streptococcal disease in Alberta, Canada- 2000-2002. AB - We determined the incidence of invasive group A streptococcal (GAS) disease among children < or =14 years of age in Alberta, Canada, with a focus on infections associated with varicella. During 2000-2002, there were 71 cases of invasive GAS disease among patients in this age group. Fourteen cases occurred in children with varicella (19.7%), for an annual incidence of 0.73 varicella-associated cases per 100,000 children. Necrotizing fasciitis was not noted in patients with invasive GAS disease and varicella. PMID- 15825002 TI - Daptomycin-resistant, methicillin-resistant Staphylococcus aureus bacteremia. AB - We describe a patient who developed daptomycin-resistant, methicillin-resistant Staphylococcus aureus (MRSA) during an episode of presumed septic thrombophlebitis of the portal vein. Although daptomycin is an alternative agent for treatment of drug-resistant gram-positive bacterial infections, development of resistance during prolonged use may occur with MRSA bacteremia from a persistent focus. PMID- 15825003 TI - The use of dexamethasone in bacterial meningitis. PMID- 15825004 TI - Is computed tomography of the head useful before lumbar puncture? PMID- 15825006 TI - Candidemia in patients with cancer: are persistent neutropenia and severity of illness score still relevant? PMID- 15825008 TI - Fatal fungemia due to Phaeoacremonium inflatipes in a child with severe aplastic anemia. PMID- 15825009 TI - Rhinovirus and lower respiratory tract infection in adults. PMID- 15825010 TI - Multidrug-resistant Shigella sonnei and Salmonella enterica Serotype typhimurium isolates producing CTX-M beta-lactamases as causes of community-acquired infection in France. PMID- 15825011 TI - Bacteremia due to Streptococcus gallolyticus, or the perils of revised nomenclature in bacteriology. PMID- 15825012 TI - Cytomegalovirus hepatitis associated with use of anti-tumor necrosis factor-alpha antibody. PMID- 15825014 TI - Cerebral tuberculomas or tuberculous brain abscess: the dilemma continues. PMID- 15825015 TI - Combination antifungal therapy for invasive aspergillosis. PMID- 15825017 TI - Successful treatment of cerebral blastomycosis with voriconazole. AB - Blastomycosis can occasionally involve the central nervous system (CNS). Amphotericin B deoxycholate is considered the drug of choice for the treatment of CNS blastomycosis. Significant toxicity may be associated with its use. We describe a case of cerebral blastomycoma that was successfully treated with voriconazole. PMID- 15825018 TI - Candida prosthetic valve endocarditis cured by caspofungin therapy without valve replacement. AB - A 64-year-old woman with a mechanical mitral valve prosthesis developed late onset Candida endocarditis. Blood cultures grew Candida glabrata and Candida krusei. Transesophageal echocardiography demonstrated vegetations on the valve. The patient was not medically fit for valve replacement, but her condition was successfully treated with 6 weeks of intravenous caspofungin therapy. PMID- 15825019 TI - Vibrio fluvialis hemorrhagic cellulitis and cerebritis. AB - We describe a case of Vibrio fluvialis hemorrhagic cellulitis and cerebritis following multiple fire-ant stings and wading in brackish water. A high index of suspicion is required for diagnosis of this specific pathogen and concordant infection. This is, to our knowledge, the first report of this type of wound infection. PMID- 15825020 TI - Modeling community- and individual-level effects of child-care center attendance on pneumococcal carriage. AB - BACKGROUND: The prevalence of pneumococcal carriage varies widely across communities. This variation is not fully explained by risk factors at the individual level but may be explained by factors producing effects at both the individual and community levels, such as child-care center (CCC) attendance. METHODS: We developed a transmission model to evaluate whether the combined risks of attending CCCs and associating with playmates who attend CCCs account for a large proportion of the variability in the prevalence of pneumococcal carriage across communities. We based parameters for the model on data from a multicommunity study. RESULTS: According to our model, differences in the proportion of children who attend CCCs can account for a range of 4%-56% in the prevalence of pneumococcal carriage across communities. Our model, which was based on data collected from 16 Massachusetts communities, predicts that the odds of carriage associated with CCC attendance are 2-3 times the odds associated with no CCC attendance (individual-level effect). The model also predicts that the odds of carriage for nonattendees in a community with CCCs are up to 6 times the odds for children in a community without CCCs (community-level effect). In addition, the mean number of hours spent at CCCs by a single attendee appears to exert effects on pneumococcal carriage that are independent of either the proportion of CCC attendance in the community or the mean number of hours these attendees spend in child care. CONCLUSIONS: We used data from multiple communities to develop a transmission model that explains marked differences in pneumococcal carriage across communities by variations in CCC attendance. This model only accounts for CCC attendance among young children and does not include other known risk factors for pneumococcal carriage. PMID- 15825021 TI - Modeling the association between pneumococcal carriage and child-care center attendance. PMID- 15825022 TI - Shigella sonnei meningitis due to interleukin-1 receptor-associated kinase-4 deficiency: first association with a primary immune deficiency. AB - BACKGROUND: Inherited interleukin-1-receptor-associated kinase-4 (IRAK-4) deficiency is a recently described immunodeficiency associated with pyogenic bacterial infections and a poor inflammatory response. Shigella sonnei is generally associated with outbreaks of rectocolitis in developed countries, but systemic illnesses have occasionally been reported. An underlying primary immunodeficiency has not been found in such cases before now. METHODS: We report the clinical and immunological features of a patient with IRAK-4 deficiency who has a history of systemic shigellosis in addition to other infections. RESULTS: The patient has a history of Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa infections during childhood and an episode of S. sonnei septicemia and meningitis at 10 years of age. This patient's history contrasted with that of other individuals infected concurrently by the same organism. Of note, these episodes were not accompanied by acute phase responses in our patient. Subsequently, the patient has had more episodes of staphylococcal disease, but no systemic illnesses. The patient is now 30 years old and has been doing well since prophylactic antibiotic treatment was stopped 4 years ago. DISCUSSION: To our knowledge, this is the first report of a case of systemic shigellosis in a person with a primary immunodeficiency, expanding the spectrum of infections associated with IRAK-4 deficiency. Thus, immunity mediated by IRAK 4 seems to be crucial for both the containment of and the inflammatory response to S. sonnei infection in the intestinal mucosa. IRAK-4 deficiency and related disorders should be considered in patients with systemic shigellosis. PMID- 15825023 TI - Variants of the natural resistance-associated macrophage protein 1 gene (NRAMP1) are associated with severe forms of pulmonary tuberculosis. AB - Although genetic factors may affect susceptibility to tuberculosis, studies that have assessed variants of the natural resistance-associated macrophage protein 1 gene (NRAMP1) and their association with tuberculosis in humans have yielded conflicting results. It is likely that NRAMP1 polymorphisms may be associated with progression to severe forms of pulmonary tuberculosis rather than with susceptibility to Mycobacterium tuberculosis infection. To test this possibility, we examined NRAMP1 variants at the INT4 and D543N loci, as well as their association with severe forms of pulmonary tuberculosis, in 127 patients with active pulmonary tuberculosis and in 91 ethnically matched, healthy control subjects in areas of China where tuberculosis is endemic. We found that NRAMP1 polymorphisms at these 2 loci were significantly associated with 2 severe forms of pulmonary tuberculosis: sputum smear-positive tuberculosis and cavitary tuberculosis. The NRAMP1 variants were not associated with pulmonary M. tuberculosis infection, when analyses of all patients with tuberculosis and all control subjects were performed. The findings of the present study support the hypothesis that genetic variants of NRAMP1 may have an effect on bacilli growth and on outcomes of pulmonary tuberculosis, but not on susceptibility to M. tuberculosis infection. PMID- 15825024 TI - Temporal-spatial analysis of severe acute respiratory syndrome among hospital inpatients. AB - BACKGROUND: We report the temporal-spatial spread of severe acute respiratory syndrome (SARS) among inpatients in a hospital ward during a major nosocomial outbreak and discuss possible mechanisms for the outbreak. METHODS: All inpatients who had stayed in the same ward as the initial index case patient for any duration before isolation were recruited into a cohort and followed up to document the occurrence of SARS. The normalized concentration of virus-laden aerosols at different locations of the ward was estimated by use of computational fluid dynamics modeling. The attack rates in the various subgroups stratified by bed location were calculated. Multivariate Cox proportional hazards regression was used to document important risk factors. RESULTS: The overall attack rate of SARS was 41% (30 of 74 subjects). It was 65%, 52%, and 18% in the same bay, adjacent bay, and distant bays, respectively (P = .001). Computation fluid dynamics modeling indicated that the normalized concentration of virus-laden aerosols was highest in the same bay and lowest in the distant bays. Cox regression indicated that staying in the ward on 6 or 10 March entailed higher risk, as well as staying in the same or adjacent bays. The epidemic curve showed 2 peaks, and stratified analyses by bed location suggested >1 generation of spread. CONCLUSIONS: The temporal-spatial spread of SARS in the ward was consistent with airborne transmission, as modeled by use of computational fluid dynamics. Infected health care workers likely acted as secondary sources in the latter phase of the outbreak. PMID- 15825025 TI - Adenovirus infection after pediatric bone marrow transplantation: is treatment always necessary? AB - BACKGROUND: Adenovirus infections are associated with significant rates of morbidity and mortality among children after bone marrow transplantation (BMT). Many transplantation units use molecular virological methods, such as polymerase chain reaction (PCR), for surveillance for adenovirus infection and give preemptive antiviral therapy to children with evidence of disseminated adenovirus infection. This treatment strategy has never been evaluated in clinical trials. METHODS: We retrospectively tested blood samples obtained from a cohort of children who had undergone BMT before the introduction of regular weekly surveillance for adenovirus infection. A total of 273 samples collected from 26 patients between May 1998 and June 2002 were tested for adenovirus infection by quantitative PCR. Virus load was quantified for each sample yielding positive test results, and the clinical notes and virological records of each child were reviewed. RESULTS: Evidence of adenovirus infection was found in 11 children (42%), 7 of whom had not previously had positive test results. Receipt of T cell depleted transplants was associated with a significantly higher incidence of adenovirus infection during the posttransplantation period. The 2 children who died from adenovirus disease developed infection within 2 weeks after transplantation, and both had very low absolute lymphocyte counts at the time of diagnosis. Seven of 11 children with blood samples that were found to be positive for adenovirus by PCR cleared the virus without antiviral therapy. CONCLUSIONS: Surveillance for adenovirus by PCR is better than symptomatic testing for detecting adenovirus infection. Antiviral therapy may not be necessary for all children who develop adenovirus viremia after BMT. PMID- 15825026 TI - Effectiveness of pneumococcal vaccination for elderly people in Catalonia, Spain: a case-control study. AB - BACKGROUND: Observational studies offer an approach to evaluating the effectiveness of vaccination programs. We evaluated the effectiveness of a 23 valent pneumococcal vaccination program for elderly people in Catalonia, Spain, in a matched-set case-control study. METHODS: We identified 149 cases of invasive pneumococcal disease among patients aged > or =65 years who were hospitalized in 12 large hospitals in Catalonia during the period of 1 January 2001 through 31 March 2002. We selected 2 hospital control patients and 1 outpatient control subject for each case patient, matching on the basis of age and underlying medical conditions. We obtained their pneumococcal vaccination histories and used conditional logistic regression to determine effectiveness of vaccination. RESULTS: Among all 149 cases of invasive pneumococcal disease, 131 (87.9%) were caused by vaccine or vaccine-related serotypes. In the adjusted analysis, overall effectiveness of vaccination against infections due to all serotypes was 70% (95% confidence interval [CI], 48%-82%). Among immunocompetent subjects with or without high-risk conditions, effectiveness of vaccination was 76% (95% CI, 51% 88%), but among immunocompromised subjects it was 50% (95% CI, -44% to 82%). Among subjects with infections due to vaccine or vaccine-related serotypes, effectiveness of vaccination was 72% (95% CI, 50%-85%) overall and 78% (95% CI, 50%-90%) in those who were immunocompetent, but it was only 46% (95% CI, -54% to 81%) in those who were immunocompromised. Overall effectiveness of vaccination was 65% (95% CI, 35%-81%) during the noninfluenza period. CONCLUSIONS: Pneumococcal vaccination was effective in preventing invasive pneumococcal disease among all elderly persons in Catalonia. Effectiveness was greater in immunocompetent persons, most of whom had underlying high-risk conditions. The number of subjects was too small to determine whether vaccination was effective in those who were immunocompromised. PMID- 15825028 TI - Colds and influenza-like illnesses in university students: impact on health, academic and work performance, and health care use. AB - BACKGROUND: Upper respiratory tract illnesses (URIs) are a major cause of morbidity among adults, with substantial direct and indirect costs to society, but their impact among university students has not been well described. We sought to assess the impact of URIs (colds and influenza-like illnesses [ILIs]) on the health, academic and work performance, and health care use of university students. METHODS: This was a cohort study of college students at the University of Minnesota, Twin Cities campus (Minneapolis-St. Paul), who were recruited during October 2002 and followed up from November 2002 through April 2003. All 42,000 registered students were invited via e-mail to participate. Baseline information was obtained in October 2002. Monthly follow-up information about colds or ILIs was obtained for the period of November 2002 through April 2003. Data were collected by use of Internet-based questionnaires. RESULTS: Of 4919 volunteers, 3249 completed all follow-up surveys. The mean age was 22.7 years; 68% of the volunteers were female. Ninety-one percent had > or = 1 URI (83% had > or = 1 cold, and 36.7% had > or = 1 ILI). These URIs caused 6023 bed-days, 4263 missed school days, 3175 missed work days, and 45,219 days of illness. Of the cohort, 22.2% had > or = 1 health care visit, and 15.8% used antibiotics to treat a URI; 27.8% did poorly on a test and 46.3% did poorly on a class assignment. ILIs versus colds had a much greater impact on all parameters (e.g., general health level was 55%-60% lower with ILI vs. no URI and 33%-39% lower for colds vs. no URI; P < .001 for each). CONCLUSION: Colds and ILIs were common and associated with substantial morbidity in university students. Enhanced efforts to prevent and control URIs, especially influenza vaccination, could improve the health and well-being of the 17 million college and university students in this country. PMID- 15825027 TI - Frequency of interleukin-4 (IL-4) -589 gene polymorphism and vaginal concentrations of IL-4, nitric oxide, and mannose-binding lectin in women with recurrent vulvovaginal candidiasis. AB - BACKGROUND: A C-->T substitution at position -589 in the interleukin-4 (IL-4) gene is associated with increased production of IL-4. Associations between this polymorphism and recurrent vulvovaginal candidiasis (RVVC), as well as vaginal concentrations of IL-4 and the anticandidal compounds nitric oxide (NO) and mannose binding lectin (MBL), were evaluated. METHODS: Vaginal samples obtained by lavage from 42 women with RVVC during the acute stage of the disease and 43 control samples were assayed by enzyme-linked immunosorbent assay for IL-4 and NO metabolites. The -589 IL-4 gene polymorphism was detected by polymerase chain reaction and endonuclease digestion. Data were analyzed by Fisher's exact test, the nonparametric Mann-Whitney and Kruskal-Wallis tests, and Spearman rank correlation. P < .05 was considered significant. RESULTS: Candida albicans was identified in 38 patients with RVVC; 3 others had infection due to Candida tropicalis, and 1 had infection due to Candida krusei. The IL-4 T,T genotype was detected in 59.5% of patients with RVVC and in 7.0% of control subjects (P < .0001). The frequency of IL-4*T was 76.2% in patients with RVVC and 23.3% in control subjects (P < .0001). The median concentration of vaginal IL-4 was elevated in patients with RVVC, compared with control subjects (P < .0001). Conversely, vaginal concentrations of NO metabolites (P = .02) and MBL (P < .0001) were reduced in patients with RVVC. There was a positive association between IL-4*T homozygosity and vaginal IL-4 levels (P < .0001) and negative associations between this genotype and vaginal NO (P = .01) and MBL (P < .0001) concentrations. CONCLUSIONS: Reduced vaginal levels of anticandidal factors in IL 4*T homozygotes may increase susceptibility to RVVC. PMID- 15825029 TI - Safety and immunogenicity of glycoprotein D-adjuvant genital herpes vaccine. AB - BACKGROUND: Two previous trials have suggested that a herpes simplex virus (HSV) type 2 glycoprotein D (gD) vaccine combined with the adjuvants alum and 3'-O deacylated-monophosphoryl lipid A (MPL) is well tolerated and provides protection against genital herpes disease in women with no preexisting HSV antibody. METHODS: The safety and immunogenicity of this vaccine were evaluated in a large, multicenter, double-blind, randomized, placebo-controlled trial. The effects of sex and preexisting HSV immunity were sought. RESULTS: When solicited symptoms that continued after the initial 4 days of observation were excluded, the incidence of unsolicited symptoms occurring during the 7 months after vaccination (the primary analysis period) was 22.1% in vaccine recipients and 21.9% in placebo recipients. Significant increases in the number of local and systemic symptoms were found in vaccine recipients within 4 days after vaccination. However, most symptoms were mild to moderate in severity and were short lived. Women reported symptoms more frequently than did men, but preexisting immunity had little effect. The vaccine induced higher titers of HSV gD antibody on enzyme linked immunosorbent assays than did natural infection with HSV. CONCLUSION: The vaccine was generally safe, well tolerated, and immunogenic. PMID- 15825030 TI - Influenza virus infection in travelers to tropical and subtropical countries. AB - BACKGROUND: Influenza outbreaks have been reported among travelers, but attack rates and incidence are unknown. METHODS: A cohort study was conducted. Travelers to subtropical and tropical countries recruited at the University of Zurich Travel Clinic (Switzerland), January 1998 to March 2000, were investigated with pre- and posttravel assessment of hemagglutination inhibition and by questionnaire. RESULTS: Among 1450 travelers recruited who completed questionnaires and provided serum samples before departure, 289 (19.9%) reported febrile illness during or after traveling abroad; of these, 211 (73.0%) provided paired serum samples. Additionally, paired serum samples were collected from 321 frequency-matched afebrile control subjects among the remaining 1161 subjects of the study population. Seroconversion for influenza virus infection was demonstrated in 40 (2.8%) of all travelers; 18 participants (1.2%) had a > or = 4 fold increase in antibody titers. This corresponds to an incidence of 1.0 influenza-associated events per 100 person-months abroad. Among the 211 febrile participants, 27 (12.8%) had seroconversion, 13 (6.2%) with a > or = 4-fold increase; among the 321 afebrile control subjects, 13 (4.0%) had seroconversion, 5 (1.6%) with a > or = 4-fold increase. Twenty-five seroconverters (62.5%; P = .747) acquired influenza outside of the European epidemic season. Sixteen patients (40.0%) sought medical attention either abroad or at home, and 32 (80.0%) were asymptomatic at the time of completion of the survey. CONCLUSIONS: This survey indicates that influenza is the most frequent vaccine-preventable infection among travelers to subtropical and tropical countries. Infections occur mainly outside the domestic epidemic season, and they have a considerable impact. Pretravel vaccination should be considered for travelers to subtropical and tropical countries. PMID- 15825031 TI - Predicting antimicrobial resistance in invasive pneumococcal infections. AB - BACKGROUND: The prevalence of multiantimicrobial resistance among Streptococcus pneumoniae continues to increase worldwide. In patients presenting with infection possibly due to pneumococci, recognition of risk factors that would identify those likely to have an antibiotic-resistant isolate might assist clinicians in choosing the most appropriate empirical therapy. METHODS: A prospective cohort study of invasive pneumococcal infection was conducted in Toronto, Canada. Risk factors for antimicrobial resistance were evaluated by means of univariate and multivariate modeling. RESULTS: A total of 3339 patients with invasive pneumococcal infection were identified between 1995 and 2002. Multivariate modeling revealed that risk factors for infection with penicillin-resistant as opposed to penicillin-susceptible pneumococci were year of infection (odds ratio [OR], 1.28; P < .001), absence of chronic organ system disease (OR, 1.72; P = .03), and previous use of penicillin (OR, 2.47; P = .006), trimethoprim sulfamethoxazole (TMP-SMX; OR, 5.97; P < .001), and azithromycin (OR, 2.78; P = .05). Infection with TMP-SMX-resistant pneumococci was associated with absence of chronic organ system disease (OR, 1.64; P = .001) and with previous use of penicillin (OR, 1.71; P = .03), TMP-SMX (OR, 4.73; P < .001), and azithromycin (OR, 3.49; P = .001). Infection with macrolide-resistant isolates was associated with previous use of penicillin (OR, 1.77; P = .03), TMP-SMX (OR, 2.07; P = .04), clarithromycin (OR, 3.93; P < .001), and azithromycin (OR, 9.93; P < .001). Infection with fluoroquinolone-resistant pneumococci was associated with previous use of fluoroquinolones (OR, 12.1; P < .001), current residence in a nursing home (OR, 12.9; P < .001), and nosocomial acquisition of pneumococcal infection (OR, 9.94; P = .003). CONCLUSIONS: Knowledge of antimicrobial use during the 3 months before infection is crucial for determining appropriate therapy for a patient presenting to the hospital with an illness for which S. pneumoniae is a possible cause. Nosocomial acquisition and nursing home acquisition are significant risk factors for infection with fluoroquinolone-resistant pneumococci. PMID- 15825032 TI - Antibiotic use: the crystal ball for predicting antibiotic resistance. PMID- 15825033 TI - Enzyme-linked immunospot assay responses to early secretory antigenic target 6, culture filtrate protein 10, and purified protein derivative among children with tuberculosis: implications for diagnosis and monitoring of therapy. AB - BACKGROUND: The ability to detect tuberculosis-specific lymphocytes by enzyme linked immunospot (ELISPOT) assay may have important implications for the diagnosis and monitoring of tuberculosis in children, for which routine methods lack sensitivity. We conducted a study to determine the presence and time course of ELISPOT responses in children with tuberculosis. METHODS: Blood samples were obtained from children with a clinical diagnosis of tuberculosis, and interferon gamma ELISPOT assays were performed using purified protein derivative (PPD), early secretory antigenic target 6 (ESAT-6), and culture filtrate protein 10 (CFP10) as stimulants. A subset of children were retested after 1, 3, and 6 months of therapy. RESULTS: Detectable responses to ESAT-6 or CFP10 were found in 49 of 70 children with clinical tuberculosis but were more frequently found in those with culture-proven disease (P = .05). The number of subjects with responses to PPD increased after 1 month of therapy (P = .0004) and decreased at 3 and 6 months. CONCLUSION: Tuberculosis-specific ELISPOT testing is a promising tool that should be evaluated as a potential diagnostic test for childhood tuberculosis. We caution against the use of an early decrease in response as a marker of successful antituberculous chemotherapy. PMID- 15825034 TI - Factors influencing the effectiveness of oseltamivir and amantadine for the treatment of influenza: a multicenter study from Japan of the 2002-2003 influenza season. AB - BACKGROUND: To evaluate the effectiveness of oseltamivir and amantadine for the treatment of influenza with respect to various clinical factors, a prospective multicenter study of the influenza season of 2002-2003 was done with 2163 patients whose condition was diagnosed by an antigen-detection test kit. METHODS: Oseltamivir was administered to 803 patients with influenza A (A+Os group) and 684 patients with influenza B (B+Os group). Amantadine was administered to 676 patients with influenza A (A+Am group). RESULTS: For each group, the duration of fever (i.e., body temperature, > or = 37.5 degrees C) was significantly shorter in patients who received the drug within 12 h after the onset of symptoms than in patients who received the drug > 12 h after the onset. For all 3 groups, the duration of fever was shorter in patients with a highest temperature < 39 degrees C than in patients with temperatures > or = 39 degrees C. The duration of fever was significantly longer for the B+Os group than for the A+Os group. Multiple regression analysis found that the type of influenza, the highest body temperature, and the time between the onset of symptoms and the start of treatment are independent factor that influence the duration of fever. CONCLUSIONS: Early administration increases the benefit of anti-influenza drugs- not only the benefit of oseltamivir treatment for influenza A, but also the benefit of amantadine treatment for influenza A and oseltamivir treatment for influenza B. Oseltamivir may be less effective as a treatment for influenza B than for influenza A. A highest body temperature of > or = 39 degrees C was an indicator of a longer duration of fever. PMID- 15825035 TI - Risk factors for increasing multidrug resistance among extended-spectrum beta lactamase-producing Escherichia coli and Klebsiella species. AB - BACKGROUND: The importance of infections due to extended-spectrum beta -lactamase producing Escherichia coli and Klebsiella species (ESBL-EK) has been increasingly recognized in recent years. ESBL-EK infections are of clinical concern, because few antimicrobials are available as therapeutic options. Increased reliance on carbapenems has led to increasing carbapenem resistance. Efforts to maintain current therapeutic options for ESBL-EK infections are essential. METHODS: We conducted a case-control study to identify risk factors for multidrug resistance (MDR) among ESBL-EK. All patients at our institution who had an inpatient clinical culture result positive for an ESBL-EK during the period of 1 June 1997 through 31 December 2002 were eligible for inclusion. An MDR ESBL-EK was defined as ESBL-EK demonstrating resistance to trimethoprim-sulfamethoxazole, aminoglycosides, and quinolones. All available ESBL-EK isolates were characterized by pulsed-field gel electrophoresis (PFGE). RESULTS: Of 361 total ESBL-EK isolates, 68 (18.8%) were MDR. During the study period, the prevalence of MDR among ESBL-EK isolates increased from 12.5% to 26.9%. The only independent risk factor for MDR ESBL-EK was the infecting organism (i.e., Klebsiella pneumoniae; adjusted odds ratio, 11.7; 95% confidence interval, 4.77-28.51; P < .001). Prior antibiotic use was not independently associated with MDR ESBL-EK. PFGE patterns from K. pneumoniae isolates indicated close genetic relatedness among a substantial proportion of isolates. CONCLUSIONS: The emergence of MDR among ESBL-EK has important implications for the future ability to treat these infections. The strong association between the species of infecting organism and MDR suggests that the epidemiology in K. pneumoniae may be unique. PFGE results suggest that horizontal spread is important in the emergence of MDR ESBL-EK. PMID- 15825036 TI - Gastroenteritis due to Listeria monocytogenes. AB - It has been known for a long time that many patients experience diarrhea antecedent to the development of bacteremia or meningoencephalitis due to Listeria monocytogenes, but it was only recently that convincing evidence was obtained that this organism can cause acute, self-limited, febrile gastroenteritis in healthy persons. At least 7 outbreaks of foodborne gastroenteritis due to L. monocytogenes have been reported. Illness typically occurs 24 h after ingestion of a large inoculum of bacteria and usually lasts 2 days. Common symptoms include fever, watery diarrhea, nausea, headache, and pains in joints and muscles. L. monocytogenes should be considered to be a possible etiology in outbreaks of febrile gastroenteritis when routine cultures fail to yield a pathogen. PMID- 15825037 TI - Colistin: the revival of polymyxins for the management of multidrug-resistant gram-negative bacterial infections. AB - The emergence of multidrug-resistant gram-negative bacteria and the lack of new antibiotics to combat them have led to the revival of polymyxins, an old class of cationic, cyclic polypeptide antibiotics. Polymyxin B and polymyxin E (colistin) are the 2 polymyxins used in clinical practice. Most of the reintroduction of polymyxins during the last few years is related to colistin. The polymyxins are active against selected gram-negative bacteria, including Acinetobacter species, Pseudomonas aeruginosa, Klebsiella species, and Enterobacter species. These drugs have been used extensively worldwide for decades for local use. However, parenteral use of these drugs was abandoned approximately 20 years ago in most countries, except for treatment of patients with cystic fibrosis, because of reports of common and serious nephrotoxicity and neurotoxicity. Recent studies of patients who received intravenous polymyxins for the treatment of serious P. aeruginosa and Acinetobacter baumannii infections of various types, including pneumonia, bacteremia, and urinary tract infections, have led to the conclusion that these antibiotics have acceptable effectiveness and considerably less toxicity than was reported in old studies. PMID- 15825038 TI - Undetectable viremia without antiretroviral therapy in patients with HIV seroconversion: an uncommon phenomenon? AB - BACKGROUND: The objective of this study was to identify the frequency and characteristics of patients with human immunodeficiency virus (HIV) infection who had spontaneously achieved viremia below the detection limit of either 400 or 500 copies/mL (depending on the test used) and to describe the duration of undetectable viremia without antiretroviral therapy (ART). METHODS: In the French Agence Nationale de Recherches sur le SIDA SEROCO cohort, 426 patients with HIV seroconversion (i.e., seroconverters) enrolled between 1988 and 1995 had serial measurement of HIV RNA levels during follow-up (with a cutoff date of 31 December 2002). Factors that distinguished those patients who had spontaneously achieved undetectable viremia (> or =2 consecutive viral loads <400 or <500 copies/mL while not receiving ART) were identified by logistic regression. A Cox model was used to estimate the predictive value of factors related to the duration of undetectable viremia. RESULTS: Undetectable viremia had been spontaneously achieved in 36 of 426 seroconverters. Women (adjusted odds ratio [aOR], 2.44; 95% confidence interval [CI], 1.03-5.80) and subjects with baseline HIV RNA level < or =3.76 log10 copies/mL (aOR, 0.31; 95% CI, 0.11-0.82), baseline HIV DNA level < or =2.61 log10 copies/mL (aOR, 0.14; 95% CI, 0.04-0.44), and high baseline CD4+ cell count (aOR, 1.18 for each 100 cells/mm3; 95% CI, 1.03-1.35]) were more likely to have achieved undetectable viremia. The sustainability of this phenomenon (median duration, 11.9 months; range, 4.6-62.8 months) was associated with low baseline HIV DNA and RNA levels. CONCLUSIONS: Achieving undetectable viremia without ART was not rare, because 6.7% of seroconverters still had a viral load of <400 or <500 copies/mL 5 years after seroconversion. These data should be considered when assessing virologic outcome for patients who interrupt highly active ART initiated during primary infection. PMID- 15825039 TI - Challenges in the study of patients with HIV type 1 seroconversion. PMID- 15825040 TI - Influence of 516G>T polymorphisms at the gene encoding the CYP450-2B6 isoenzyme on efavirenz plasma concentrations in HIV-infected subjects. AB - We examined 516G>T polymorphisms at the gene encoding the cytochrome P450 in 100 human immunodeficiency virus-positive subjects who were receiving efavirenz (EFV). Elevated plasma EFV concentrations were found in 40% of subjects with the polymorphic homozygous genotype and 19% of subjects with the heterozygous genotype. Conversely, 20% of subjects with the wild-type genotype had subtherapeutic concentrations of EFV. CYP2B6-516 genotyping may help to identify subjects who have plasma EFV concentrations that are outside of the therapeutic range. PMID- 15825041 TI - Influence of ciprofloxacin resistance on risk factors for endovascular infection in patients with infection due to group C nontyphoid salmonellae. AB - From January 2000 through September 2004, a total of 54 patients with infection due to group C nontyphoid salmonellae were evaluated; 8 patients had gastroenteritis alone, and 46 patients had bacteremia. Of the 46 patients who had bacteremia, 12 had endovascular infection and 34 did not. The number of infections due to ciprofloxacin-resistant Salmonella organisms is increasing. Ciprofloxacin-resistant Salmonella organisms predisposed patients to acquire bacteremia, but they did not seem to predispose patients to acquire endovascular infection. PMID- 15825042 TI - Characterization and management of paradoxical upgrading reactions in HIV uninfected patients with lymph node tuberculosis. AB - In a retrospective analysis, paradoxical deterioration of lymph node tuberculosis in human immunodeficiency virus-uninfected patients was common, occurring in 25 (23%) of 109 patients after treatment had been given for a median of 46 days (interquartile range, 21-139 days) and persisting for a median of 67.5 days (interquartile range, 34-111 days). We found no association between the use of steroids and the duration of reaction. PMID- 15825043 TI - Issues pertaining to data extraction and classification and publication bias in meta-analysis of the diagnostic accuracy of markers for bacterial infection. PMID- 15825045 TI - Important factors to consider for patients with community-acquired pneumonia. PMID- 15825047 TI - Appropriate antimicrobial therapy for community-acquired methicillin-resistant Staphylococcus aureus carrying the Panton-Valentine leukocidin genes. PMID- 15825049 TI - How infrequent are opportunistic diseases and immune reconstitution syndromes among HIV-infected individuals who have favorable CD4+ cell count responses to antiretroviral therapy? PMID- 15825051 TI - High risk of infectious disease caused by salmonellae and mycobacteria infections in patients with Crohn disease treated with anti-interleukin-12 antibody. PMID- 15825052 TI - Anti-interleukin-12 antibody treatment for Crohn disease: potential risk of invasive disease due to mycobacteria and salmonellae infection. PMID- 15825053 TI - Mismanagement of malaria among travelers in developing countries. PMID- 15825055 TI - Clostridium difficile infection among health care workers receiving antibiotic therapy. PMID- 15825059 TI - NIH slated for annual increase in 2006 of less than one percent. PMID- 15825062 TI - Certificate of added qualification for hepatology: vindication not vanity. PMID- 15825063 TI - Image of the month. PMID- 15825064 TI - Increased risk of rectal cancer after prostate radiation: a population-based study. AB - BACKGROUND & AIMS: Radiation therapy for prostate cancer has been associated with an increased rate of pelvic malignancies, particularly bladder cancer. The association between radiation therapy and colorectal cancer has not been established. METHODS: We conducted a retrospective cohort study using Surveillance, Epidemiology, and End Results (SEER) registry data from 1973 through 1994. We focused on men with prostate cancer, but with no previous history of colorectal cancer, treated with either surgery or radiation who survived at least 5 years. We evaluated the effect of radiation on development of cancer for 3 sites: definitely irradiated sites (rectum), potentially irradiated sites (rectosigmoid, sigmoid, and cecum), and nonirradiated sites (the rest of the colon). Using a proportional hazards model, we evaluated the effect of radiation on development of colorectal cancer over time. RESULTS: A total of 30,552 men received radiation, and 55,263 underwent surgery only. Colorectal cancers developed in 1437 patients: 267 in irradiated sites, 686 in potentially irradiated sites, and 484 in nonirradiated sites. Radiation was independently associated with development of cancer over time in irradiated sites but not in the remainder of the colon. The adjusted hazards ratio for development of rectal cancer was 1.7 for the radiation group, compared with the surgery-only group (95% CI: 1.4-2.2). CONCLUSIONS: We noted a significant increase in development of rectal cancer after radiation for prostate cancer. Radiation had no effect on development of cancer in the remainder of the colon, indicating that the effect is specific to directly irradiated tissue. PMID- 15825065 TI - Trichuris suis therapy for active ulcerative colitis: a randomized controlled trial. AB - BACKGROUND & AIMS: Ulcerative colitis is most common in Western industrialized countries. Inflammatory bowel disease is uncommon in developing countries where helminths are frequent. People with helminths have an altered immunological response to antigens. In animal models, helminths prevent or improve colitis by the induction of regulatory T cells and modulatory cytokines. This study determined the efficacy and safety of the helminth Trichuris suis in therapy of ulcerative colitis. METHODS: This was a randomized, double blind, placebo controlled trial conducted at the University of Iowa and select private practices. Trichuris suis ova were obtained from the US Department of Agriculture. The trial included 54 patients with active colitis, defined by an Ulcerative Colitis Disease Activity Index of > or =4. Patients were recruited from physician participants and were randomly assigned to receive placebo or ova treatment. Patients received 2500 Trichuris suis ova or placebo orally at 2-week intervals for 12 weeks. RESULTS: The primary efficacy variable was improvement of the Disease Activity Index to > or =4. After 12 weeks of therapy, improvement according to the intent-to-treat principle occurred in 13 of 30 patients (43.3%) with ova treatment compared with 4 of 24 patients (16.7%) given placebo (P = .04). Improvement was also found with the Simple Index that was significant by week 6. The difference in the proportion of patients who achieved an Ulcerative Colitis Disease Activity Index of 0-1 was not significant. Treatment induced no side effects. CONCLUSIONS: Ova therapy seems safe and effective in patients with active colitis. PMID- 15825066 TI - Sippy of the Sippy diet regimen. PMID- 15825068 TI - Fertility and pregnancy-related events in women with celiac disease: a population based cohort study. AB - BACKGROUND & AIMS: Previous studies have raised concern about reduced fertility and increased adverse pregnancy-related events in women with celiac disease, but none has estimated overall fertility compared with the general female population. METHODS: We compared computerized primary care data for 1521 women with celiac disease with data for 7732 age- and practice-matched women without celiac disease. We estimated population-based rates of fertility and adverse pregnancy outcomes. RESULTS: Crude fertility rates were 48.2 and 47.7 live births per 1000 person-years for women with and without celiac disease, respectively (rate ratio, 1.01; 95% confidence interval, 0.90-1.14). Age-specific fertility rates showed that women with celiac disease had lower fertility when younger but higher fertility when older compared with women without celiac disease. This increase in relative fertility with increasing age held whether women had treated or untreated celiac disease. Risks of cesarean section (odds ratio, 1.33; 95% confidence interval, 1.03-1.70) and miscarriage (rate ratio, 1.31; 95% confidence interval, 1.06-1.61) were moderately higher in women with celiac disease, but risks of assisted birth, breech birth, preeclampsia, postpartum hemorrhage, ectopic pregnancy, stillbirth, and termination were similar. CONCLUSIONS: Overall, women with celiac disease have fertility similar to that of the general female population, but they have their babies at an older age. Although our findings may reflect a disease effect, the age shift in fertility rates and the increase in cesarean section risk is consistent with socioeconomic or educational advantages of women with celiac disease. PMID- 15825067 TI - Duodenal ulcer promoting gene of Helicobacter pylori. AB - BACKGROUND & AIMS: Identification of a disease-specific H pylori virulence factors predictive of the outcome of infection remains unachieved. METHODS: We used the polymerase chain reaction and Southern blot to compare the presence of 14 vir homologue genes with clinical presentation of H pylori infection, mucosal histology, and mucosal interleukin (IL)-8 levels. RESULTS: We examined 500 H pylori strains from East Asia and South America, including 120 with gastritis, 140 with duodenal ulcer (DU), 110 with gastric ulcer (GU), and 130 with gastric cancer. Only 1 gene that encompassed both jhp0917 and jhp0918 called dupA (duodenal ulcer promoting gene) was associated with a specific clinical outcome. dupA was present in 42% of DU vs. 21% of gastritis (adjusted odds ratio [OR] = 3.1, 95% confidence interval [CI]: 1.7-5.7). Its presence was also associated with more intense antral neutrophil infiltration and IL-8 levels and was a marker for protection against gastric atrophy, intestinal metaplasia, and gastric cancer (OR for gastric cancer = 0.42, 95% CI: 0.2-0.9 compared with gastritis). In vitro studies in gastric epithelial cells using dupA -deleted and -complemented mutants showed that the dupA plays roles in IL-8 production, in activation of transcription factors responsible for IL-8 promoter activity, and in increased survivability at low pH. CONCLUSIONS: dupA is a novel marker associated with an increased risk for DU and reduced risk for gastric atrophy and cancer. Its association with DU-promoting and -protective effects against atrophy/cancer was evident in both Asian and Western countries. PMID- 15825069 TI - Ornidazole for prophylaxis of postoperative Crohn's disease recurrence: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND & AIMS: Crohn's disease almost inevitably recurs after ileocolonic resection, and effective prophylactic therapy has not been identified. We investigated the efficacy and safety of ornidazole, a nitroimidazole antibiotic, for the prevention of clinical recurrence of Crohn's disease after curative ileocolonic resection in a placebo-controlled double-blind clinical trial. METHODS: Eighty patients were randomized to ornidazole 1 g/day or placebo started within 1 week of resection and continued for 1 year. The primary end point was the proportion of patients with clinical recurrence at 1 year. Secondary end points were endoscopic recurrence at 3 months and 12 months after resection. RESULTS: Two patients in the ornidazole group withdrew consent and were not dosed. Ornidazole significantly reduced the clinical recurrence rate at 1 year from 15 of 40 (37.5%) patients in the placebo group to 3 of 38 (7.9%) patients in the ornidazole group (Fisher exact test, 8.03; P = .0046; odds ratio, 0.14; 95% confidence interval, 0.037-0.546). Ornidazole reduced endoscopic recurrence at 12 months from 26 of 33 (79%) in the placebo group to 15 of 28 (53.6%) in the ornidazole group (chi2 , 4.37; P = .037; odds ratio, 0.31; 95% confidence interval, 0.10-0.94). Endoscopic recurrence at 3 and 12 months predicted clinical recurrence. Significantly more patients in the ornidazole group dropped out from the study because of side effects (P = .041). CONCLUSIONS: Ornidazole 1 g/day is effective for the prevention of recurrence of Crohn's disease after ileocolonic resection. PMID- 15825070 TI - Infliximab maintenance treatment reduces hospitalizations, surgeries, and procedures in fistulizing Crohn's disease. AB - BACKGROUND & AIMS: Infliximab is effective in closing fistulas in patients with Crohn's disease. We examined the effect of infliximab maintenance treatment on hospitalizations, surgeries, and procedures in patients with fistulizing Crohn's disease enrolled in the ACCENT II study. METHODS: After 5 mg/kg infliximab at weeks 0, 2, and 6, a total of 282 patients were separately randomized at week 14 as responders (at least a 50% reduction from baseline in the number of draining fistulas at both weeks 10 and 14) or nonresponders to receive placebo or 5 mg/kg infliximab maintenance every 8 weeks. At week 22 and later, patients who lost response could be treated with a maintenance dose 5 mg/kg higher. Data on Crohn's disease-related hospitalizations, surgeries, and procedures were compared between the treatment groups for responders and all randomized patients. RESULTS: A total of 282 patients were randomized at week 14, of whom 195 were randomized as responders. Among patients randomized as responders, those who received infliximab maintenance had significantly fewer mean hospitalization days (0.5 vs. 2.5 days; P < .05), mean numbers (per 100 patients) of hospitalizations (11 vs. 31; P < .05), all surgeries and procedures (65 vs. 126; P < .05), inpatient surgeries and procedures (7 vs. 41; P < .01), and major surgeries (2 vs. 11; P < .05), compared with those who received placebo maintenance. CONCLUSIONS: In patients with fistulizing Crohn's disease, infliximab 5 mg/kg every 8 weeks significantly reduced hospitalizations, surgeries, and procedures compared with placebo. PMID- 15825071 TI - Randomized study comparing banding and propranolol to prevent initial variceal hemorrhage in cirrhotics with high-risk esophageal varices. AB - BACKGROUND & AIMS: Standard care for prevention of first esophageal variceal hemorrhage is beta-blockade, but this may be ineffective or unsafe. Our purpose was to compare endoscopic banding with propranolol for prevention of first variceal hemorrhage. METHODS: In a multicenter, prospective trial, 62 patients with cirrhosis with high-risk esophageal varices were randomized to propranolol (titrated to reducing resting pulse by > or =25%) or banding (performed monthly until varices were eradicated) and were followed up on the same schedule for a mean duration of 15 months. The primary end point was treatment failure, defined as the development of endoscopically documented variceal hemorrhage or a severe medical complication requiring discontinuation of therapy. Direct costs were estimated from Medicare reimbursements and fixed or variable charges for services up to treatment failure. RESULTS: Background variables of the treatment groups were similar. The trial was stopped early after an interim analysis showed that the failure rate of propranolol was significantly higher than that of banding (6/31 vs. 0/31; difference, 19.4%; P = .0098; 95% confidence interval for true difference, 6.4%-37.2%). Significantly more propranolol than banding patients had esophageal variceal hemorrhage (4/31 vs. 0/31; difference, 12.9%; P = .0443; 95% confidence interval for true difference, 0.8%-29%), and the cumulative mortality rate was significantly higher in the propranolol than in the banding group (4/31 vs. 0/31; difference, 12.9%; P = .0443; 95% confidence interval for true difference, 0.8%-29%). Direct costs of care were not significantly different. CONCLUSIONS: For patients with cirrhosis with high-risk esophageal varices and no history of variceal hemorrhage, propranolol-treated patients had significantly higher failure rates of failure, first esophageal varix hemorrhage, and cumulative mortality than banding patients. Direct costs of medical care were not significantly different. PMID- 15825072 TI - Colchicine treatment of alcoholic cirrhosis: a randomized, placebo-controlled clinical trial of patient survival. AB - BACKGROUND & AIMS: Colchicine improved survival and reversed cirrhosis in several small clinical trials. We compared the efficacy and safety of long-term colchicine, as compared with placebo, in patients with advanced alcoholic cirrhosis. METHODS: Five hundred forty-nine patients with advanced (Pugh B or C) alcoholic cirrhosis were randomized to receive either colchicine 0.6 mg twice per day (n = 274) or placebo (n = 275). Treatment lasted from 2 to 6 years. The primary outcome was all-cause mortality. Secondary outcomes were liver-related morbidity and mortality. Liver biopsy was requested prior to entry and after 24 months of treatment. RESULTS: Attendance at scheduled clinic visits and adherence with study medication were similar in colchicine and placebo groups. Alcohol intake was less than 1 drink per day in 69% of patients. In an intention-to-treat analysis, all-cause mortality was similar in colchicine (49%) and placebo (45%) patients (P = .371). Mortality attributed to liver disease was 32% in colchicine and 28% in placebo patients (P = .337). Fewer patients receiving colchicine developed hepatorenal syndrome. In 54 patients with repeat liver biopsies after 24 or more months of treatment, cirrhosis improved to septal fibrosis in 7 patients (3 colchicine, 4 placebo) and to portal fibrosis in 1 patient (colchicine). CONCLUSIONS: In patients with advanced alcoholic cirrhosis, colchicine does not reduce overall or liver-specific mortality. Liver histology improves to septal fibrosis in a minority of patients after 24 months of treatment, with similar rates of improvement in patients receiving placebo and colchicine. Colchicine is not recommended for patients with advanced alcoholic cirrhosis. PMID- 15825073 TI - Variable phenotypes of enterocolitis in interleukin 10-deficient mice monoassociated with two different commensal bacteria. AB - BACKGROUND & AIMS: To explore the hypothesis that selective immune responses to distinct components of the intestinal microflora induce intestinal inflammation, we characterized disease kinetics and bacterial antigen-specific T-cell responses in ex germ-free interleukin 10 -/- and wild-type control mice monoassociated with Enterococcus faecalis , Escherichia coli , or Pseudomonas fluorescens . METHODS: Colitis was measured by using blinded histological scores and spontaneous interleukin 12 secretion from colonic strip culture supernatants. Interferon gamma secretion was measured from mesenteric or caudal lymph node CD4 + T cells stimulated with bacterial lysate-pulsed antigen-presenting cells. Luminal bacterial concentrations were measured by culture and quantitative polymerase chain reaction. RESULTS: Escherichia coli induced mild cecal inflammation after 3 weeks of monoassociation in interleukin 10 -/- mice. In contrast, Enterococcus faecalis-monoassociated interleukin 10 -/- mice developed distal colitis at 10-12 weeks that was progressively more severe and associated with duodenal inflammation and obstruction by 30 weeks. Neither bacterial strain induced inflammation in wild-type mice, and germ-free and Pseudomonas fluorescens monoassociated interleukin 10 -/- mice remained disease free. CD4 + T cells from Enterococcus faecalis- or Escherichia coli-monoassociated interleukin 10 -/- mice selectively produced higher levels of interferon gamma and interleukin 4 when stimulated with antigen-presenting cells pulsed with the bacterial species that induced disease; these immune responses preceded the onset of histological inflammation in Enterococcus faecalis -monoassociated mice. Luminal bacterial concentrations did not explain regional differences in inflammation. CONCLUSIONS: Different commensal bacterial species selectively initiate immune-mediated intestinal inflammation with distinctly different kinetics and anatomic distribution in the same host. PMID- 15825074 TI - Pancreatic carcinoma cells induce fibrosis by stimulating proliferation and matrix synthesis of stellate cells. AB - BACKGROUND & AIMS: Tumor desmoplasia is one of the representative histopathologic findings in ductal pancreatic adenocarcinoma. The aims of this study were to examine the cellular and molecular mechanisms of fibrogenesis associated with pancreatic adenocarcinomas. METHODS: Immunostainings were performed with human pancreatic adenocarcinomas (n = 27) and tumors induced in nude mice (n = 36) by subcutaneously injecting MiaPaCa2, Panc1, and SW850 with and without pancreatic stellate cells. Matrix-producing cells were isolated from pancreatic adenocarcinomas and compared with pancreatic stellate cells isolated from tissue of chronic pancreatitis. Paracrine stimulation of pancreatic stellate cells by carcinoma cells was studied regarding matrix synthesis (collagen and c fibronectin on protein and messenger RNA level) and cell proliferation (bromodeoxyuridine incorporation). RESULTS: High numbers of desmin and alpha smooth muscle actin-positive cells were detected in 26 of 27 pancreatic adenocarcinomas. Intense fibronectin and collagen stainings were associated with these cells. By using cytofilament stainings, gene expression profiling, and morphological examinations, the matrix-producing cells obtained by the outgrowth method from pancreatic adenocarcinomas were identified as pancreatic stellate cells. Supernatants of MiaPaCa2, Panc1, and SW850 cells stimulated proliferation and collagen type I and c-fibronectin synthesis of cultured pancreatic stellate cells. Preincubation of the carcinoma cell supernatants with neutralizing antibodies against fibroblast growth factor 2, transforming growth factor beta 1, and platelet-derived growth factor significantly reduced the stimulatory effects. Subcutaneous injection of carcinoma cells and pancreatic stellate cells induced fast-growing subcutaneous fibrotic tumors in nude mice. Morphometric analysis of carcinoma cells (cytokeratin stainings) showed a high density of carcinoma cells in these tumors. CONCLUSIONS: Pancreatic stellate cells strongly support tumor growth in the nude mouse model. The increased deposition of connective tissue in pancreatic carcinoma is the result of a paracrine stimulation of pancreatic stellate cells by carcinoma cells. PMID- 15825075 TI - Colitis in mice lacking the common cytokine receptor gamma chain is mediated by IL-6-producing CD4+ T cells. AB - BACKGROUND & AIMS: Mice that have a truncated mutation of the common cytokine receptor gamma chain (CR gamma -/Y) are known to spontaneously develop colitis. To identify the pathologic elements responsible for triggering this localized inflammatory disease, we elucidated and characterized aberrant T cells and their enteropathogenic cytokines in CR gamma -/Y mice with colitis. METHODS: The histologic appearance, cell population, T-cell receptor V beta usage, and cytokine production of lamina propria lymphocytes were assessed. CR gamma -/Y mice were treated with anti-interleukin (IL)-6 receptor monoclonal antibody to evaluate its ability to control colitis, and splenic CD4 + T cells from the same mouse model were adoptively transferred into SCID mice to see if they spurred the appearance of colitis. RESULTS: We found marked thickening of the large intestine, an increase in crypt depth, and infiltration of the colonic lamina propria and submucosa with mononuclear cells in the euthymic CR gamma -/Y mice, but not in the athymic CR gamma -/Y mice, starting at the age of 8 weeks. Colonic CD4 + T cells with high expressions of antiapoptotic Bcl-x and Bcl-2 were found to use selected subsets (V beta 14) of T-cell receptor and to exclusively produce IL-6. Treatment of CR gamma -/Y mice with anti-IL-6 receptor monoclonal antibody prevented the formation of colitis via the induction of apoptosis in IL-6 producing CD4 + T cells. Adoptive transfer of pathologic CD4 + T cells induced colitis in the recipient SCID mice. CONCLUSIONS: Colonic IL-6-producing thymus derived CD4 + T cells are responsible for the development of colitis in CR gamma /Y mice. PMID- 15825076 TI - Loss of Klf4 in mice causes altered proliferation and differentiation and precancerous changes in the adult stomach. AB - BACKGROUND & AIMS: The epithelial zinc-finger transcription factor Klf4 (formerly GKLF) regulates cellular proliferation and differentiation in vitro. Klf4 null mice die by postnatal day 1 and show changes in epithelial differentiation of skin and colon. METHODS: We used tissue-specific gene ablation to generate mice lacking Klf4 in their gastric epithelia. Klf4 mutant mice and controls were killed for histology, immunohistochemistry, quantitative real-time polymerase chain reaction (qPCR), and serum gastrin levels. Klf4 messenger RNA (mRNA) levels were analyzed in Foxa3-Cdx2 transgenic mice and controls. Human gastric cancers and matched normal tissue were used for qPCR and immunohistochemistry for KLF4. RESULTS: Klf4 mutant mice survive to adulthood and show increased proliferation and altered differentiation of their gastric epithelia. Klf4 mutants also display aberrant expression of acidic mucins and TFF2/SP-positive cells, findings characteristic of premalignant conditions, but no inflammation, intestinal metaplasia, dysplasia, or cancer up to 1 year of age. Expression of KLF4 is nearly absent in human gastric cancer, suggesting that failure to activate KLF4 during normal cellular differentiation may be a common feature of gastric cancers. p21 WAF1/CIP1 is an in vivo target of Klf4, but Klf4 is not a mediator of Cdx2. CONCLUSIONS: Loss of a single genetic factor, Klf4, leads to dramatic changes in the gastric epithelia of mice, and Klf4 is part of a regulatory pathway involving p21 WAF1/CIP1 but not Cdx2. Thus, Klf4 is critical for normal gastric epithelial homeostasis. PMID- 15825077 TI - CDX2-regulated expression of iron transport protein hephaestin in intestinal and colonic epithelium. AB - BACKGROUND & AIMS: The homeobox transcription factor CDX2 has a key role in intestinal development and differentiation. Mice heterozygous for Cdx2 inactivation develop colonic polyps with epithelial cells showing gastric or squamous differentiation. Loss of CDX2 expression is seen in some poorly differentiated colon carcinomas in humans. Conversely, ectopic CDX2 expression in the stomach of transgenic mice promotes intestinal metaplasia, and CDX2 expression often is seen in intestinal metaplasia in stomach and esophagus. To enhance knowledge of CDX2 function, we sought to define CDX2-regulated genes. METHODS: HT-29 and WiDr colorectal cancer (CRC) cells with low endogenous CDX2 expression were transduced with a CDX2 expression vector, and gene expression changes were assessed by microarrays. RESULTS: The gene for ceruloplasmin-related iron transport protein hephaestin (HEPH) was induced by CDX2 in HT-29 and WiDr. In other CRC lines and human and mice tissues, endogenous HEPH expression was linked to CDX2 expression. Activation of CDX2 rapidly induced HEPH expression, and RNA interference-mediated inhibition of CDX2 led to lower HEPH expression. Studies with HEPH reporter gene constructs and chromatin-immunoprecipitation approaches suggested that CDX2 directly regulates HEPH transcription. In CRC cells, CDX2 induction suppressed intracellular iron levels, consistent with the view that HEPH regulates iron export. CDX2 expression was modulated in response to changes in intracellular iron levels, implying a regulatory pathway in which increased iron levels lead to increased expression of CDX2 and HEPH and enhanced iron export. CONCLUSIONS: CDX2 has a key role in regulating HEPH expression and iron levels in intestinal cells. PMID- 15825078 TI - Serotonin inhibits Na+/H+ exchange activity via 5-HT4 receptors and activation of PKC alpha in human intestinal epithelial cells. AB - BACKGROUND & AIMS: Increased serotonin levels have been implicated in the pathophysiology of diarrhea associated with celiac and inflammatory diseases. However, the effects of serotonin on Na+ /H+ exchange (NHE) activity in the human intestine have not been investigated fully. The present studies examined the acute effects of 5-hydroxytryptamine (5-HT) on NHE activity using Caco-2 cells as an in vitro model. METHODS: Caco-2 cells were treated with 5-HT (.1 micromol/L, 1 h) and NHE activity was measured as ethyl-isopropyl-amiloride (EIPA)-sensitive 22Na uptake. The effect of 5-HT receptor-specific agonists and antagonists was examined. The role of signaling intermediates in 5-HT-mediated effects on NHE activity was elucidated using pharmacologic inhibitors and immunoblotting. RESULTS: NHE activity was inhibited significantly (approximately 50%-75%, P < .05) by .1 micromol/L 5-HT via inhibition of maximal velocity (Vmax) without any changes in apparent affinity (Km) for the substrate Na+ . NHE inhibition involved a decrease of both NHE2 and NHE3 activities. Studies using specific inhibitors and agonists showed that the effects of 5-HT were mediated by 5-HT4 receptors. 5 HT-mediated inhibition of NHE activity was dependent on phosphorylation of phospholipase C gamma 1 (PLC gamma 1) via activation of src-kinases. Signaling pathways downstream of PLC gamma 1 involved increase of intracellular Ca 2+ levels and subsequent activation of protein kinase C alpha (PKC alpha). The effects of 5-HT on NHE activity were not cell-line specific because T84 cells also showed NHE inhibition. CONCLUSIONS: A better understanding of the regulation of Na+ absorption by 5-HT offers the potential for providing insights into molecular and cellular mechanisms involved in various diarrheal and inflammatory disorders. PMID- 15825079 TI - Therapeutic effects of rectal administration of basic fibroblast growth factor on experimental murine colitis. AB - BACKGROUND & AIMS: Basic fibroblast growth factor (bFGF) is a promising therapeutic agent for various diseases. It remains unclear, however, whether bFGF is effective for the treatment of inflammatory bowel disease. The aim of this study was to examine the efficacy of bFGF on 2 experimental murine colitis models and to investigate its molecular mechanisms. METHODS: We evaluated the effects of human recombinant bFGF (hrbFGF) on mice with dextran sulfate sodium (DSS)-induced colitis and mice with trinitrobenzene sulfonic acid (TNBS)-induced colitis as well as normal mice. Body weight, survival rate, and histologic findings of the colonic tissues were examined. Gene expression of tumor necrosis factor (TNF) alpha, cyclooxygenase (COX)-2, transforming growth factor (TGF)-beta, mucin 2 (MUC2), intestinal trefoil factor (ITF), and vascular endothelial growth factor (VEGF) in the colonic tissues was determined. The proliferation activity of hrbFGF on the colonic epithelium was evaluated by immunohistochemistry. RESULTS: Rectal administration of hrbFGF ameliorated DSS-induced colitis in a dose dependent manner. Gene expression of TNF-alpha was significantly reduced in the colonic tissues of mice with DSS-induced colitis treated with hrbFGF, whereas MUC2 and ITF messenger RNA expression was up-regulated. Rectal administration of hrbFGF significantly improved the survival rate of mice with TNBS-induced colitis and partially ameliorated colitis. hrbFGF significantly increased the number of Ki-67-positive cells in the colonic epithelium of normal mice, and up-regulated the gene expression of COX-2, TGF-beta, MUC2, ITF, and VEGF in the colonic tissues. CONCLUSIONS: Rectal administration of bFGF might be a promising option for the treatment of inflammatory bowel disease. PMID- 15825081 TI - Clostridium difficile toxin B activates the EGF receptor and the ERK/MAP kinase pathway in human colonocytes. AB - BACKGROUND & AIMS: Clostridium difficile toxin B (TxB) mediates acute inflammatory diarrhea characterized by neutrophil infiltration and intestinal mucosal injury. In a xenograft animal model, TxB was shown to induce interleukin (IL)-8 gene expression in human colonic epithelium. However, the precise mechanisms of this TxB response are unknown. The aim of this study was to investigate the TxB-mediated proinflammatory pathway in colonocytes. METHODS: The effect of TxB on epidermal growth factor receptor (EGFR), extracellular signal regulated kinase (ERK) 1/2 signaling pathway and IL-8 gene expression was assessed in nontransformed human colonic epithelial NCM460 cells. TxB regulation of EGFR-ERK1/2 signaling pathways was determined using immunoblot analysis, confocal microscopy, and enzyme-linked immunosorbent assay, whereas IL-8 gene expression was measured by luciferase promoter assay. RESULTS: TxB activates EGFR and ERK1/2 phosphorylation with subsequent release of IL-8 from human colonocytes. Pretreatment with either the EGFR tyrosine kinase inhibitor, AG1478, or an EGFR-neutralizing antibody blocked both TxB-induced EGFR and ERK activation. By using neutralizing antibodies against known ligands of EGFR, we found that the activation of EGFR and ERK1/2 phosphorylation was mediated by transforming growth factor-alpha (TGF-alpha). Inhibition of matrix metalloproteinase (MMP) decreased TGF-alpha secretion and TxB-induced EGFR and ERK activation. Inhibition of MMP, EGFR, and ERK activation significantly decreased TxB-induced IL-8 expression. CONCLUSIONS: TxB signals acute proinflammatory responses in colonocytes by transactivation of the EGFR and activation of the ERK/MAP kinase pathway. PMID- 15825080 TI - Distinct temporal-spatial roles for rho kinase and myosin light chain kinase in epithelial purse-string wound closure. AB - BACKGROUND & AIMS: Small epithelial wounds heal by purse-string contraction of an actomyosin ring that is regulated by myosin light chain (MLC) kinase (MLCK) and rho kinase (ROCK). These studies aimed to define the roles of these kinases in purse-string wound closure. METHODS: Oligocellular and single-cell wounds were created in intestinal epithelial monolayers. Fluorescence imaging and electrophysiologic data were collected during wound closure. Human biopsies were studied immunohistochemically. RESULTS: Live-cell imaging of enhanced green fluorescent protein-beta-actin defined rapid actin ring assembly within 2 minutes after wounding. This progressed to a circumferential ring within 8 minutes that subsequently contracted and closed the wound. We therefore divided this process into 2 phases: ring assembly and wound contraction. Activated rho and ROCK localized to the wound edge during ring assembly. Consistent with a primary role in the assembly phase, ROCK inhibition prevented actin ring assembly and wound closure. ROCK inhibition after ring assembly was complete had no effect. Recruitment and activation of MLCK occurred after ring assembly was complete and coincided with ring contraction. MLCK inhibition slowed and then stopped contraction but did not prevent ring assembly. MLCK inhibition also delayed barrier function recovery. Studies of human colonic biopsy specimens suggest that purse-string wound closure also occurs in vivo, because MLC phosphorylation was enhanced surrounding oligocellular wounds. CONCLUSIONS: These results suggest complementary roles for these kinases in purse-string closure of experimental and in vivo oligocellular epithelial wounds; rho and ROCK are critical for actin ring assembly, while the activity of MLCK drives contraction. PMID- 15825082 TI - Increased expression and function of integrins in enterocytes by endotoxin impairs epithelial restitution. AB - BACKGROUND & AIMS: Experimental necrotizing enterocolitis (NEC) is characterized by circulating endotoxin (lipopolysaccharide [LPS]) and impaired enterocyte migration. We hypothesized that LPS increases integrin function and cell-matrix adhesion, leading to impaired enterocyte migration in the pathogenesis of NEC. METHODS: NEC-like intestinal injury was induced in newborn rats by hypoxia/gavage feedings, and restitution was determined by assessing bromodeoxyuridine-labeled enterocytes along the crypt-villus axis. Newborn mice were injected with 5 mg/kg LPS. IEC-6 cells were treated with LPS +/- LY294002 or wortmannin, and beta 1- and alpha 3-integrins were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunofluorescence. Beta 1-integrin function was determined by adherence of fibronectin beads to IEC-6 monolayers. Migration of IEC-6 cells into a scraped wound was measured by time-lapse microscopy. RESULTS: Newborn intestinal injury was associated with decreased intestinal restitution and increased alpha 3- and beta 1-integrin expression in the ileal mucosa, which also was observed after LPS injection. In IEC-6 cells, LPS caused an increase in the expression of alpha 3- and beta 1-integrins, a shift of beta 1-integrins from the cytoplasm to the plasma membrane and an increase in fibronectin bead adhesion during which beta 1-integrins accumulated underneath attached beads. These effects could be reversed with LY294002 or wortmannin, suggesting phosphatidylinositol-3-phosphate kinase (PI3K) dependence. The increased integrin matrix adhesion by LPS led to an inhibition of enterocyte migration, which could be reversed by anti-beta 1-antibodies. CONCLUSIONS: Enterocyte migration is inhibited by LPS through increased expression and function of alpha 3- and beta 1 integrins. Modulation of enterocyte migration via integrins may provide novel insights into the pathogenesis of NEC, in which intestinal restitution is impaired. PMID- 15825083 TI - Identification of cholelithogenic enterohepatic helicobacter species and their role in murine cholesterol gallstone formation. AB - BACKGROUND & AIMS: Helicobacter spp are common inhabitants of the hepatobiliary and gastrointestinal tracts of humans and animals and cause a variety of well described diseases. Recent epidemiologic results suggest a possible association between enterohepatic Helicobacter spp and cholesterol cholelithiasis, chronic cholecystitis, and gallbladder cancer. To test this, we prospectively investigated the effects of Helicobacter spp infection in cholesterol gallstone pathogenesis in the highly susceptible C57L/J mouse model. METHODS: Helicobacter spp-free adult male C57L mice were infected with several different enterohepatic Helicobacter spp or left uninfected and fed either a lithogenic diet or standard mouse chow for 8 and 18 weeks. At the conclusion of the study, bile was examined microscopically and diagnostic culture and polymerase chain reaction were performed. RESULTS: Mice infected with Helicobacter bilis or coinfected with Helicobacter hepaticus and Helicobacter rodentium and fed a lithogenic diet developed cholesterol gallstones at 80% prevalence by 8 weeks compared with approximately 10% in uninfected controls. Monoinfections with H hepaticus , Helicobacter cinaedi , and H rodentium gave a cholesterol gallstone prevalence of 40%, 30%, and 20%, respectively; the latter 2 groups did not differ significantly from uninfected animals. Neither infected nor uninfected mice fed a chow diet developed cholesterol gallstones. CONCLUSIONS: These findings, along with prior epidemiologic studies, suggest that Helicobacter spp play a major role in the pathophysiology of cholesterol gallstone formation in mice and perhaps humans. PMID- 15825084 TI - Hepatitis C virus expression suppresses interferon signaling by degrading STAT1. AB - BACKGROUND & AIMS: The molecular mechanisms by which hepatitis C virus (HCV) antagonizes the antiviral actions of interferon (IFN) have not been fully characterized. Specifically, how HCV proteins impact on IFN signaling components has yet to be elucidated. We used an HCV cell-based expression model to examine the interaction between HCV protein expression and host type I IFN signaling components in the Jak-STAT kinase pathway. METHODS: Full-length HCV and HCV subgenomic constructs corresponding to structural and each of the nonstructural proteins were transiently transfected into Huh-T7 cells. HCV expression was monitored by an HCV core antigen enzyme-linked immunosorbent assay. STAT1, P STAT1, and HCV protein expression was investigated with immunoprecipitation and Western blots. RESULTS: Overexpression and small interfering RNA studies showed that STAT1 was indispensable for control of HCV expression. STAT1 and P-STAT1 expression were markedly reduced in HCV-transfected cells. Full-length HCV, HCV core/E1/E2, and NS3-4A were each associated with decreased STAT1 expression, which was attributable to proteasome-dependent degradation of STAT1. HCV core, but not HCV E1, E2, NS3, NS4, or NS5, bound to STAT1. STAT2 expression was not affected by HCV. CONCLUSIONS: HCV expression selectively degrades STAT1 and reduces P-STAT1 accumulation in the nucleus in a proteasome-dependent manner. HCV core protein binds STAT1, suggesting that this viral protein is associated with STAT1 degradation. STAT1 plays an indispensable role in innate antiviral immunity against HCV expression. In turn, HCV subverts the Jak-STAT kinase by selectively inducing STAT1 degradation. PMID- 15825085 TI - Bile acids induce hepatic stellate cell proliferation via activation of the epidermal growth factor receptor. AB - BACKGROUND & AIMS: Hepatic stellate cell (HSC) proliferation is a key event in the development of liver fibrosis. In many liver diseases, HSCs are exposed to inflammatory cytokines, reactive oxygen species, and bile acids. Although inflammatory cytokines and reactive oxygen species are known to promote proliferation of HSCs, nothing is known about the effects of bile acids on HSC proliferation or apoptosis. The aim of this study was to investigate the effects of bile acids on HSC proliferation. METHODS: HSCs were exposed to bile acids with different hydrophobicity (5-200 micromol/L). HSC proliferation and cell cycle related events were assessed by bromodeoxyuridine incorporation, cell counting and proliferating cell nuclear antigen and cyclin E expression, apoptosis by caspase-3 activity assay, immunocytochemistry for active caspase-3 and acridine orange staining, and activation of signal transduction pathways by Western blot using phospho-specific antibodies. Uptake of bile acids was investigated using fluorescent bile acids. RESULTS: All bile acids, at concentrations >25 micromol/L, induce a 2.5- to 3-fold increase in HSC proliferation via activation of the epidermal growth factor receptor. Bile acid-induced proliferation is mediated by activation of a protein kinase C/extracellular signal-regulated kinase/p70S6K-dependent pathway. Bile acids did not induce apoptosis in HSCs. HSCs do not take up fluorescent bile acids and do not express the bile acid importer ntcp. CONCLUSIONS: Bile acids at levels reached in cholestatic conditions are an independent profibrogenic factor. Bile acids induce HSC proliferation via the activation of the epidermal growth factor receptor, whereas HSCs are protected against bile acid-induced apoptosis by excluding bile acids. PMID- 15825086 TI - Mathematical modeling of primary hepatitis C infection: noncytolytic clearance and early blockage of virion production. AB - BACKGROUND & AIMS: Although hepatitis C virus kinetics and immune determinants during primary infection have been described, the virus-host interplay is not fully understood. We used mathematical modeling to elucidate and quantify virus host dynamics. METHODS: Ten chimpanzees were infected intrahepatically with H77 RNA (n = 3) or intravenously with infected serum. Blood samples were taken 1-3 times per week for 6 months. A new model was fitted to the observed HCV RNA and alanine aminotransferase (ALT) kinetics. RESULTS: After infection, viral levels increased in a biphasic manner with a transient decline in between. This can be explained by a partial block (mean, 91%) of virion production, possibly due to an endogenous type I interferon response. After reaching maximum levels, a long viral plateau (mean, 6.1 log cp/mL) can be explained by blind homeostasis and lack of susceptible cells. Modest elevations in ALT levels (21-93 IU/L) were concurrently observed, indicating a shorter half-life of infected versus noninfected hepatocytes (mean ratio, 2.6). Following the ALT flare, viral titers rapidly declined to a lower (mean, 4.5 log cp/mL; n = 6) or undetectable level (n = 4). This decline is compatible with increased cell death (mean minimal estimate half-life, 28.7 days) and noncytolytic clearance (mean maximal estimate half life, 24.1 days) of infected cells. CONCLUSIONS: Our results quantify virus-host dynamics during primary HCV infection and suggest that endogenous type I interferon slows virus production in the early acute phase. Partial or effective virus control correlates with the half-life of infected cells regulated by both cytolytic and noncytolytic mechanisms. PMID- 15825087 TI - Colorectal cancer at a young age. PMID- 15825088 TI - Oral contraceptives and polyp regression in familial adenomatous polyposis. AB - Epidemiologic and experimental reports suggest that female hormones protect against the development of colorectal cancer, but studies are limited. We describe a patient in the placebo arm of a 4-year primary chemoprevention trial who developed adenomatous polyps and then had eradication of polyps after the administration of oral contraceptives. No change in the prostaglandin levels in the colonic mucosa was noted after polyp elimination, making nonsteroidal anti inflammatory drug ingestion unlikely as a cause. This report represents the regression of colorectal adenomas with the use of estrogen/progesterone compounds. PMID- 15825090 TI - Gastrointestinal food allergy: new insights into pathophysiology and clinical perspectives. AB - Adverse reactions to food that result in gastrointestinal symptoms are common in the general population; while only a minority of such individuals will have symptoms due to immunologic reactions to foods, gastrointestinal food allergies do exist in both children and adults. These immune reactions are mediated by immunoglobulin E-dependent and -independent mechanisms involving mast cells, eosinophils, and other immune cells, but the complexity of the underlying mechanisms of pathogenesis have yet to be fully defined. Knowledge of the spectrum of adverse reactions to foods that affect the digestive system, including gastrointestinal food allergy, is essential to correctly diagnose and manage the subset of patients with immunologically mediated adverse reactions to foods. Potentially fatal reactions to food necessitate careful instruction and monitoring on the part of health care workers involved in the care of individuals at risk of anaphylaxis. New methods of diagnosis and novel strategies for treatment, including immunologic modulation and the development of hypoallergenic foods, are exciting developments in the field of food allergy. PMID- 15825089 TI - Gene expression profiling of intestinal epithelial cell maturation along the crypt-villus axis. AB - BACKGROUND & AIMS: To define the genetic reprogramming that drives intestinal epithelial cell maturation along the crypt-villus axis, enterocytes were sequentially isolated from the villus tip to the crypts of mouse small intestine. METHODS: Changes in gene expression were assessed using 27,405-element complementary DNA microarrays (14,685 unique genes) and specific changes validated by Western blotting. RESULTS: A total of 1113 genes differentially expressed between the crypt and villus were identified. Among these, established markers of absorptive and goblet cell differentiation were up-regulated in villus cells, whereas Paneth cell markers were maximally expressed in crypt cells. The 1113 differentially expressed genes were significantly enriched for genes involved in cell cycle progression, RNA processing, and translation (all predominantly down-regulated during maturation) and genes involved in cytoskeleton assembly and lipid uptake (predominantly up-regulated during maturation). No enrichment for apoptosis-regulating genes was observed. We confirmed that Wnt signaling was maximal in the proliferative compartment and observed a decrease in MYC and an increase in MAD and MAX expression during the maturation program. Consistent with these changes, the 1113 genes were enriched for MYC targets, establishing the importance of this network in intestinal cell maturation. CONCLUSIONS: This database serves as a resource for understanding the molecular mechanisms of intestinal cell maturation and for dissection of how perturbations in the maturation process can lead to changes in gastrointestinal physiology and pathology, particularly intestinal tumorigenesis. PMID- 15825091 TI - Ionizing radiation and rectal cancer: victims of our own success. PMID- 15825092 TI - A novel approach to the treatment of ulcerative colitis: is it kosher? PMID- 15825093 TI - Primary prophylaxis for variceal bleeding: are we there yet? PMID- 15825094 TI - Commensal flora: wolf in sheep's clothing. PMID- 15825096 TI - Does acid suppression cause community-acquired pneumonia? PMID- 15825095 TI - Helicobacter and cholesterol gallstones: do findings in the mouse apply to man? PMID- 15825097 TI - Colon ischemia--need to unravel its predisposition. PMID- 15825098 TI - Organ failure as an indicator of severity of acute pancreatitis: time to revisit the Atlanta classification. PMID- 15825099 TI - IgG food antibodies and irritating the bowel. PMID- 15825100 TI - Early HVPG measurement and tips for acute variceal hemorrhage: is sooner really better? PMID- 15825105 TI - Hypothermia may impair hepatic regeneration in acute liver failure. PMID- 15825107 TI - Caution before embracing serum markers of liver fibrosis in clinical practice. PMID- 15825108 TI - Biomarkers as a first-line estimate of injury in chronic liver diseases: time for a moratorium on liver biopsy? PMID- 15825112 TI - Should balloon dilatation of the sphincter oddi be avoided in clinical practice? PMID- 15825113 TI - Recombinant factor VIIa for variceal bleeding: when, why, and how? PMID- 15825115 TI - National Institutes of Health Consensus Development Conference Statement on Celiac Disease, June 28-30, 2004. AB - NIH consensus and state-of-the-science statements are prepared by independent panels of health professionals and public representatives on the basis of (1) the results of a systematic literature review prepared under contract with the Agency for Healthcare Research and Quality (AHRQ); (2) presentations by investigators working in areas relevant to the conference questions during a 2-day public session; (3) questions and statements from conference attendees during open discussion periods that are part of the public session; and (4) closed deliberations by the panel during the remainder of the second day and morning of the third. This statement is an independent report of the panel and is not a policy statement of the NIH or the Federal Government. The statement reflects the panel's assessment of medical knowledge available at the time the statement was written. Thus, it provides a "snapshot in time" of the state of knowledge on the conference topic. When reading the statement, keep in mind that new knowledge is inevitably accumulating through medical research. PMID- 15825116 TI - Overview and pathogenesis of celiac disease. PMID- 15825117 TI - Should adults be screened for celiac disease? What are the benefits and harms of screening? AB - The symptoms of celiac disease are diverse, and the disease is often asymptomatic. Without active serologic screening, most cases probably remain undiagnosed. Recent serologic screening assays allow mass screening for the disease. However, there is no evidence as yet to suggest that symptom-free celiac disease patients run an increased risk of small intestinal lymphoma or other complications. The prevention of osteoporosis seems to be the strongest indicator for widespread screening today. Screening asymptomatic individuals for celiac disease may be even harmful. A lifelong gluten-free diet is not easy to maintain, and the subject's quality of life may deteriorate. It is also debatable whether patients found by active screening adhere to a gluten-free diet similarly to symptomatic ones. The cost-effectiveness of population screening is dubious. Serologic screening should be applied in individuals with even subtle symptoms indicative of celiac disease, such as subclinical-isolated iron deficiency. In various autoimmune conditions, the risk of celiac disease is approximately 5% and, in individuals with affected first-degree relatives, 15%. Infertility, neurologic symptoms such as polyneuropathy, ataxia, epilepsy with posterior cerebral calcification, and osteoporosis are conditions in which celiac disease should be kept in mind. Elevated aminotransferases and liver failure can lead to a diagnosis of celiac disease. Evidence today does not support mass screening of celiac disease. Instead, increased alertness should be observed in patients at risk of the condition. PMID- 15825118 TI - Consequences of testing for celiac disease. AB - Population screening studies have identified that up to two thirds of celiac disease (CD) cases are asymptomatic. The aim of this study was to conduct a systematic review of the expected consequences of testing for CD in the following populations: (1) patients with symptoms suggestive of CD, (2) asymptomatic at risk populations, and (3) general population. Standard systematic review methodology was used. A comprehensive literature search was conducted in MEDLINE (1996-2003), EMBASE (1974-2003), CAB (1972 forward), PsychINFO (1840-2003), AGRICOLA (1970-2003), and Sociological Abstracts (1963 forward); searches were conducted in December 2003. Pooled summary estimates were not calculated. The majority of the included studies were before-after studies, case control, or retrospective cohorts. The quality of evidence for the before-after studies is weaker. The overall strength of the evidence for this issue was fair to good. This area of research is relatively new, and further high-quality studies are required. The consequences of testing for celiac disease in symptomatic individuals appears to have a positive impact on patient-relevant outcomes. The data are less clear for those with silent CD or those with lower grade histologic lesions in small bowel biopsy. The literature suggests that compliance is less than ideal in these individuals, especially if diagnosed when adults. Long-term outcomes have not been extensively studied in those with silent CD. PMID- 15825119 TI - Dietary guidelines and implementation for celiac disease. AB - Medical nutrition therapy is the only accepted treatment for celiac disease. This paper summarizes a review of scientific studies using the gluten-free diet, nutritional risk factors, controversial elements of the diet, and its implementation in treating celiac disease. Treatment for celiac disease requires elimination of the storage proteins found in wheat, rye, and barley. The inclusion of oats and wheat starch is controversial. Research supports that oats may be acceptable for patients with celiac disease and can improve the nutritional quality of the diet. However, use of oats is not widely recommended in the United States because of concerns of potential contamination of commercial oats. Studies assessing the contamination of commercial oats are limited. Research indicates no differences in patients choosing a strict wheat starch containing, gluten-free diet vs. a naturally gluten-free diet. Factors other than trace gluten may be the cause of continued villous atrophy in some patients. The impact of nutrient malabsorption caused from untreated celiac disease is well documented. The diet and gluten-free products are often low in B vitamins, calcium, vitamin D, iron, zinc, magnesium, and fiber. Few gluten-free products are enriched or fortified, adding to the risk of nutrient deficiencies. Patients newly diagnosed or inadequately treated have low bone mineral density, imbalanced macronutrients, low fiber intake, and micronutrient deficiencies. Also troubling is the increased incidence of obesity seen in persons with celiac disease following a gluten-free diet. Because of the nutritional risks associated with celiac disease, a registered dietitian must be part of the health care team that monitors the patient's nutritional status and compliance on a regular basis. PMID- 15825120 TI - The gluten-free diet: how to provide effective education and resources. AB - A strict gluten-free diet (GFD) for life is the only treatment for celiac disease (CD). This article reviews (1) the impact of the GFD on the quality of life of individuals with CD and their families; (2) the causes of poorly controlled CD; (3) the access to and source and quality of information provided by health professionals and other groups; (4) management strategies, including nutritional assessment and education guidelines; (5) a variety of resources available to individuals and health professionals; (6) innovative educational initiatives and partnerships; and (7) specific recommendations to address the increasing numbers of people with CD and the growing need for gluten-free (GF) foods and further education about CD and the GFD. Successful management of CD requires a team approach, including the person with CD and his or her family, physician, dietitian, and celiac support group; an individualized approach; understanding of quality of life issues; use of evidence-based, current information and resources; and regular follow-up to monitor compliance, nutritional status, and additional information and support. The physician must clearly communicate, with a positive attitude, an overview of CD and strongly emphasize the importance of a GFD for life. It is essential that the physician initiate an immediate referral to a dietitian with expertise in CD for nutritional assessment, diet education, meal planning, and assistance with the adaptation to the challenging new gluten-free lifestyle. Good dietary compliance will reduce the risk of further complications and associated health care costs and improve quality of life in patients with CD. PMID- 15825121 TI - Follow-up of patients with celiac disease: achieving compliance with treatment. AB - Celiac disease is the only autoimmune condition for which we know the environmental trigger: gluten. Complete removal of gluten from the diet in a patient with celiac disease should result in symptomatic, serologic, and histologic remission. However, compliance with the gluten-free diet, especially in the United States, is extremely challenging. Compliance can be measured both noninvasively, by dietary history and measurement of serum antibodies, and invasively, by using endoscopic and histologic criteria. The advantages and disadvantages of these various modalities are discussed. The highest rates of compliance are reported in patients who are diagnosed as young children, whereas adolescents and those diagnosed via mass serologic screening have the most transgressions. Barriers to compliance include the poor palatability of gluten free foods, confusing food-labeling practices, and common comorbid psychologic burdens such as anxiety and depression. Because celiac disease is a multisystemic disorder, physicians need to be aware of the potential autoimmune, nutritional, and malignant complications. An algorithm for the follow-up and management of the newly diagnosed celiac disease patient is presented, which includes regular follow-up; measurement of serum antibodies; eliciting a detailed dietary history; and examination for signs and symptoms of nutritional deficiencies, malignancy, and other autoimmune diseases. Ideally, a team approach to the follow-up of the newly diagnosed patient should include regular supervision by an interested physician, medical nutritional counseling by a registered dietician, and access to local and national support groups knowledgeable about this condition. PMID- 15825122 TI - Clinical features and diagnosis of celiac disease. AB - Celiac disease is a chronic enteropathy caused by intolerance to gluten. The true prevalence of this condition is much greater than previously recognized, with increasing numbers of silent cases being diagnosed. Population-based studies, using serologic screening, have indicated that the prevalence of celiac disease in Caucasian populations is .5%-1%. The pattern of incidence is changing, with a greater proportion of cases diagnosed later in adulthood. The pathologic lesion is characterized by a flattened small intestinal mucosa with a lymphocytic infiltrate, crypt hyperplasia, and villous atrophy. Absorptive function may be impaired and patients can experience gastrointestinal symptoms and malabsorption leading to development of anemia, osteoporosis, or other complications. Untreated celiac disease is associated with significant morbidity and increased mortality, largely owing to the development of enteropathy-associated intestinal lymphoma. The pathologic changes and symptoms resolve when gluten is excluded from the diet for a sustained period. PMID- 15825123 TI - What are the sensitivity and specificity of serologic tests for celiac disease? Do sensitivity and specificity vary in different populations? AB - A number of serologic tests are available commercially for identifying individuals who require an intestinal biopsy examination to diagnose celiac disease (CD). The aim of this study was to determine which test, or combination of tests, was most sensitive and specific for this purpose. We performed a literature review of studies that determined the sensitivity and specificity of serologic tests for CD. Studies that compared biopsy examination-confirmed cases of CD with controls with normal intestinal histology were selected for analysis. Sensitivities and specificities for the antigliadin tests were highly variable. Immunoglobulin (Ig)G-based antigliadin (AGA) tests generally were poor in both parameters whereas the IgA-based test was poorly sensitive but more specific. The IgA endomysium (EMA-IgA) and tissue transglutaminase (TTG-IgA) tests were both highly sensitive and specific with values for both parameters exceeding 95% in most studies. There were no identifiable differences between adults and children with respect to these tests. There was no evidence that a combination of tests was better than a single test using either the EMA IgA or TTG IgA. Either the EMA IgA or TTG-IgA test is most useful for identifying individuals with CD. The variability and generally lower accuracy associated with the AGA tests make them unsuitable for screening purposes. There is no advantage to using a panel of tests as opposed to a single test. Because these data were obtained largely from studies conducted in a research setting, it is possible the tests will be less accurate when used in the clinical setting. PMID- 15825124 TI - Genetic testing: who should do the testing and what is the role of genetic testing in the setting of celiac disease? AB - Celiac disease is a remarkable and common immune-mediated disorder determined by both the presence of characteristic HLA alleles (DQ2 and DQ8) and one of the best characterized environmental factors (gliadin) for any common autoimmune disease. The discovery of transglutaminase autoantibodies and the development of assays for these antibodies has allowed the identification of a large number of asymptomatic individuals with autoimmunity and intestinal biopsy evidence of celiac lesions. Further understanding of the sequelae of asymptomatic celiac disease, and the interaction between genetic susceptibility and environmental factors, are likely to alter fundamentally both genetic screening for celiac disease and its therapy. PMID- 15825125 TI - The diagnostic accuracy of serologic tests for celiac disease: a systematic review. AB - Clinicians are increasingly utilizing noninvasive serologic tests for the diagnosis and screening of celiac disease (CD). The aim of this study was to conduct a systematic review of the diagnostic performance of serologic tests for the diagnosis and screening of CD. Standard systematic review methodology was used. A literature search was conducted in MEDLINE (1966 to October 2003) and EMBASE (1974 to December 2003) databases. A weighted mean of the sensitivity and specificity along with 95% confidence intervals and summary receiver operating characteristic (ROC) curves were calculated. The pooled specificity of endomyseal antibody (EMA)-monkey esophagus (ME) or EMA-human umbilical cord (HU) was close to 100% in adults and children. The pooled specificity of transglutaminase antibody (tTG)-guinea pig (GP) and tTG-human recombinant (HR) were between 95% and 99%. IgA-EMA-ME demonstrated sensitivities of 96% and 97% in children and adults, respectively. EMA-HU demonstrated a similar sensitivity of 97% in children but 90% in adults. The pooled sensitivity of tTG-GP in adults and children was 90% and 93%, respectively. The sensitivity of tTG-HR was 98% and 96%, respectively. The performance of antigliadin antibody was inferior to that of EMA and tTG. EMA and tTG offer high sensitivity and specificity. The sensitivity of these tests appears to be lower than reported when milder histologic grades are used to define CD (below 90%). If true, the nearly perfect negative predictive value of these tests would drop. The positive predictive value of these tests is likely lower than reported when the tests are applied in low-prevalence populations. PMID- 15825126 TI - Epidemiology of celiac disease: what are the prevalence, incidence, and progression of celiac disease? PMID- 15825127 TI - Celiac disease in patients with an affected member, type 1 diabetes, iron deficiency, or osteoporosis? AB - Celiac disease (CD) may be a much under recognized condition, in part because it is not considered in those patients at high risk or in clinical conditions that may be a manifestation of CD. Screening high-risk groups may reveal a higher prevalence than seen in the general population. Several diseases have been described to have an association with CD that may be greater than by chance alone. The presence of autoimmune conditions such as insulin-dependent diabetes mellitus (T1DM) or a family history of CD or dermatitis herpetiformis may increase the risk of coexisting CD. The prevalence of CD may be increased in certain patient groups, including the following: osteoporosis or low bone mass or iron-deficiency anemia. Many studies, including population-based work, suggest a prevalence of CD of 3%-7% of T1DM and 4%-10% of first-degree family members. Anemia and osteoporosis are common in patients with newly diagnosed CD. Conversely, CD is common in referral populations with those conditions but makes a relatively small contribution to the overall community prevalence of anemia or postmenopausal osteoporosis. Most screen-found patients tend to have little or no gastrointestinal symptoms. In conclusion, significant reservoirs of CD can be found in some at-risk groups, such as those with T1DM, family members, and referred patients with osteoporosis and anemia. It is not clear what impact CD has on the quality of life of these individuals. PMID- 15825128 TI - The prevalence of celiac disease in average-risk and at-risk Western European populations: a systematic review. AB - Until recently, celiac disease (CD) was felt to be a rare disease in the United States. The aim of this study was to conduct a systematic review of the prevalence of CD in general Western populations and in populations at high risk for CD. Standard systematic review methodology was used. A literature search was conducted in MEDLINE (1966 to October 2003) and EMBASE (1974 to December 2003) databases. Qualitative and quantitative prevalence estimates were produced after assessing study heterogeneity. The prevalence of CD in general Western populations is close to 1% and is somewhat higher in certain Western European populations. The prevalence of CD in populations at risk for CD is as follows: 3% 6% in type 1 diabetic patients, up to 20% in first-degree relatives, 10%-15% in symptomatic iron-deficiency anemia (IDA), 3%-6% in asymptomatic IDA, and 1%-3% in osteoporosis. The prevalence of CD in patients suspected of having CD varied depending on the reasons for suspecting CD and on whether the study was conducted in a referral center. In general, the prevalence ranged from 5% to 15%, but was up to 50% in symptomatic patients evaluated in a tertiary referral center. CD is a common medical condition. The prevalence is higher still in high-risk groups. Clinicians in a variety of specialties should have a high index of suspicion for the diagnosis of CD and in particular need to pay close attention to the identified high-risk groups. PMID- 15825129 TI - Clinical presentation of celiac disease in the pediatric population. AB - Celiac disease (CD) is an immune-mediated enteropathy triggered by the ingestion of gluten-containing grains (including wheat, rye, and barley) in genetically susceptible individuals. CD is associated with HLA molecules DQ2 (90%-95%) and DQ8 (5%-10%), and in the continued presence of gluten the disease is self perpetuating. CD is one of the most common lifelong disorders worldwide and is characterized by a variety of clinical presentations. These include the typical malabsorption syndrome (classic symptoms) and a spectrum of symptoms potentially affecting any organ or body system (nonclassic symptoms). Because CD often is atypical or even clinically silent, many cases go undiagnosed and are exposed to the risk of long-term complications. There is growing interest in the social aspects of CD because the burden of illness related to this condition is doubtless higher than previously thought. PMID- 15825130 TI - The many faces of celiac disease: clinical presentation of celiac disease in the adult population. AB - The major modes of presentation of patients with celiac disease are the classic diarrhea-predominant form and silent celiac disease. Those with silent celiac disease lack diarrhea, although they may present with manifestations of celiac disease that include an irritable bowel syndrome, anemia, osteoporosis, neurologic diseases, or malignancy. A significant proportion of patients are diagnosed through screening at-risk groups including relatives of patients and insulin-dependant diabetics. Nondiarrheal presentations now are seen more commonly than those with diarrhea. Patients with celiac disease have a greater burden of disease than the general population because of autoimmune diseases and malignancies. There is a need for screening studies of patients with conditions associated with celiac disease to determine whether the large numbers of people with undiagnosed celiac disease currently are seeking health care. PMID- 15825131 TI - Association of celiac disease and intestinal lymphomas and other cancers. AB - Celiac disease (CD) is associated with intestinal lymphoma and other forms of cancer, especially adenocarcinoma of the small intestine, of the pharynx, and of the esophagus. Enteropathy-associated T-cell lymphoma (EATL) is a rare form of high-grade, T-cell non-Hodgkin lymphoma (NHL) of the upper small intestine that is specifically associated with CD. This NHL subtype arises in patients with either previously or concomitantly diagnosed CD. In a subgroup of patients, there is progressive deterioration of a refractory form of CD. EATL derives from a clonal proliferation of intraepithelial lymphocytes and is often disseminated at diagnosis. Extraintestinal presentations are not uncommon in the liver/spleen, thyroid, skin, nasal sinus, and brain. The outlook of EATL is poor. Recent studies indicated that (1) CD is associated with a significantly increased risk for NHL, especially of the T-cell type and primarily localized in the gut (EATL); (2) the CD-lymphoma association is less common than previously thought, with a relative risk close to 3; (3) CD screening is not required in patients with NHL of any primary site at the onset, unless suggested by specific findings (T-cell origin and/or primary gut localization). The risk of NHL associated with clinically milder (or silent) forms could be lower than in typical cases of CD. Several follow-up studies suggest that the GFD protects from cancer development, especially if started during the first years of life. Strict adherence to the GFD seems to be the only possibility of preventing a subset of rare but very aggressive forms of cancer. PMID- 15825132 TI - Skin manifestations of celiac disease. PMID- 15825133 TI - Neurologic presentation of celiac disease. AB - Celiac disease (CD) long has been associated with neurologic and psychiatric disorders including cerebellar ataxia, peripheral neuropathy, epilepsy, dementia, and depression. Earlier reports mainly have documented the involvement of the nervous system as a complication of prediagnosed CD. However, more recent studies have emphasized that a wider spectrum of neurologic syndromes may be the presenting extraintestinal manifestation of gluten sensitivity with or without intestinal pathology. These include migraine, encephalopathy, chorea, brain stem dysfunction, myelopathy, mononeuritis multiplex, Guillain-Barre-like syndrome, and neuropathy with positive antiganglioside antibodies. The association between most neurologic syndromes described and gluten sensitivity remains to be confirmed by larger epidemiologic studies. It further has been suggested that gluten sensitivity (as evidenced by high antigliadin antibodies) is a common cause of neurologic syndromes (notably cerebellar ataxia) of otherwise unknown cause. Additional studies showed high prevalence of gluten sensitivity in genetic neurodegenerative disorders such as hereditary spinocerebellar ataxia and Huntington's disease. It remains unclear whether gluten sensitivity contributes to the pathogenesis of these disorders or whether it represents an epiphenomenon. Studies of gluten-free diet in patients with gluten sensitivity and neurologic syndromes have shown variable results. Diet trials also have been inconclusive in autism and schizophrenia, 2 diseases in which sensitivity to dietary gluten has been implicated. Further studies clearly are needed to assess the efficacy of gluten-free diet and to address the underlying mechanisms of nervous system pathology in gluten sensitivity. PMID- 15825134 TI - Should all children be screened for celiac disease? AB - To address the issue of screening children for celiac disease, current evidence has been summarized and placed within the context of 8 established criteria for childhood screening. Screening all children for celiac disease is not recommended at this time. Areas with gaps in knowledge are identified as areas in need of further study. These include the timing of screening, defining the natural history of screening-identified celiac disease, developing tools to predict disease onset and disease remission, and the risks of screening. PMID- 15825137 TI - Fever and fussiness in a 17-day-old infant. PMID- 15825138 TI - Illnesses among recently immigrated children. AB - The number of children immigrating to the United States has increased steadily during the last decade. American families are adopting a significant portion of these children, more than 20,000. Recently immigrated children face many different health risks when compared to children born in the United States. They are subject to many infectious diseases no longer seen commonly in the United States such as malaria, tuberculosis, and HIV. They are more likely to have inadequate immunity to vaccine-preventable illnesses. Recent immigrants have a higher likelihood of having malnutrition and developmental delay. Finally, many will have suffered psychological trauma in either institutions or refugee camps. These children require specialized testing, care, and treatment in the pediatric office. PMID- 15825139 TI - Respiratory tract infections in children in developing countries. AB - Acute respiratory infections are the main cause of morbidity worldwide, and pneumonia represents one of the main causes of death in children younger than the age of 5 years in developing countries. Several risk factors for acquiring respiratory infections in developing countries, such as poverty, restricted family income, low parental education level, low birth weight, malnutrition, and lack of breastfeeding, have been described. Another important factor in recent years that has aggravated the problem of acute respiratory infections in developing countries is the human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) pandemic. In developing countries, identifying the etiology is difficult and World Health Organization recommends making the diagnosis of pneumonia based on clinical parameters. The diagnosis and suitable treatment administered by primary healthcare workers, particularly in rural areas, is part of the strategy to prevent mortality. Finally, it is important that industrialized countries support the economic growth of the developing countries to improve their healthcare system and to ensure that performed research studies will give benefits to their populations. PMID- 15825140 TI - Current concepts of childhood tuberculosis. AB - Tuberculosis (TB) is a social disease with medical implications. An estimated one third of the world's population (2 billion people) is infected with the tubercle bacilli. In the year 2000 alone, an estimated 8.3 million new cases of TB occurred, of which 884,019 (10.7%) were in children. Of the total, 659,379 (75%) occurred in 22 high-burden countries, most of which were resource-poor. Despite a growing body of literature demonstrating the substantial impact made by addressing childhood TB, current guidelines are geared toward the treatment and control of TB in adults. This review focuses on TB disease among children younger than 15 years of age, the standard World Health Organization category for TB in children. The purpose of this work is to review the epidemiology, pathophysiology, and clinical presentation of childhood TB. The challenges of accurate surveillance, confirmation of diagnoses, and effective treatment of childhood TB in resource-poor settings are highlighted. PMID- 15825141 TI - Pediatric malaria in the developing world. AB - Hundreds of millions of people suffer from malaria, and more than a million children die of malaria each year. Malaria typically presents with fever and headache, but the presentation often is nonspecific. The diagnosis should be based on blood tests, and thick and thin smears are the standard means of identifying parasites. In some areas, chloroquine still is effective as treatment, but other medications are needed in most parts of the world. Patients with severe disease (altered consciousness, marked anemia, and/or respiratory distress) should begin therapy parenterally. Control measures depend on the use of insecticide-treated bednets, early identification and treatment of symptomatic individuals, and intermittent preventive therapy. Progress continues toward the development of a useful vaccine. PMID- 15825142 TI - Treatment of human immunodeficiency virus (HIV) in children using antiretroviral drugs. AB - Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) is having a devastating impact on African men, women and children. Antiretroviral treatment of children has lagged behind that of adults in Africa and globally. Fortunately, several national and international initiatives are helping to catalyze access of HIV-infected children to treatment. In general, the principles of antiretroviral treatment are the same for resource-rich and resource-poor settings. However, the more rapid progression of HIV disease often observed among children in Africa and some other resource-poor settings may argue for a more aggressive approach to initiation of treatment. In addition, numerous barriers to treatment of HIV-infected children in Africa and other resource-poor settings exist and must be overcome, including the expense of antiretroviral medications, lack of pediatric drug formulations, and poor human capacity and infrastructure for treatment administration. The 2.2 million African children currently living with HIV/AIDS, and many more living in poor countries on other continents, are dependent on all of us to work creatively to overcome barriers to the large-scale implementation of programs for health-restoring, life-prolonging antiretroviral treatment. PMID- 15825143 TI - A millennium update on pediatric diarrheal illness in the developing world. AB - More than one billion diarrhea episodes occur every year among children younger than 5 years of age in socioeconomically developing countries causing 2 to 2.5 million deaths. More than twenty viral, bacterial, and parasitic enteropathogens are currently associated with acute diarrhea. Rotavirus and diarrheagenic Escherichia coli are the most common pathogens responsible for acute diarrhea episodes in children; Shigella spp., Salmonella spp, Campylobacter jejuni/coli, Vibrio cholerae, Aeromonas spp, and Plesiomonas spp. occur more commonly in poorer areas and infections caused by protozoa and helminthes occur mainly in areas where environmental sanitation is significantly deteriorated. Initial clinical assessment of a child with diarrhea should focus on obtaining an accurate evaluation of hydration and nutritional status. Assessment of stool characteristics (e.g., liquid non-bloody stools vs. dysenteric or bloody stools) is a key feature in determining potential pathogens causing an acute diarrhea episode. Diagnostic guidelines are discussed in the article. The major therapeutic intervention for all individuals with diarrhea consists of fluid and electrolyte therapy. When antimicrobial therapy is appropriate, selection of a specific agent should be made based upon susceptibility patterns of the pathogen or information on local susceptibility patterns. Current guidelines for administering appropriate antimicrobial treatment are provided in the article. Preventive measures include careful personal hygiene, especially promotion of hand washing. Immunizations currently or soon to be available for Salmonella serotype Typhi, cholera prevention, and rotavirus are discussed. PMID- 15825144 TI - Gnathostomiasis: a review of a previously localized zoonosis now crossing numerous geographical boundaries. AB - Expansion in international travel and increases in immigration have resulted in an increased number of persons in Europe and other Western countries who have returned from foreign travel with parasitic infections rarely seen previously in the United States. Among the diseases caused by helminthic parasites is gnathostomiasis, a disease caused by Gnathostoma spp. Once confined primarily to Southeast Asia, it now is a public health concern in Mexico and other countries. This article reviews the causes and epidemiology of gnathostomiasis, the life cycle of the helminth, the clinical picture and diagnosis of gnathostomiasis, and the treatments for and means of avoiding the disease. PMID- 15825145 TI - Alfred Fabian Hess (1875-1933). 1955. PMID- 15825147 TI - Activation of human T lymphocytes under conditions similar to those that occur during exposure to microgravity: a proteomics study. AB - A number of experiments, conducted under microgravity conditions, i.e. in space shuttle biolaboratories or in ground based systems simulating the conditions occurring in microgravity, show that in hypogravity, in vitro human lymphocyte activation is severely impaired. However, very early stimulation steps of T lymphocytes are not compromised, since CD69 receptor, the earliest membrane activation marker, is expressed by T cells at a level comparable to that observed on 1 g activated lymphocytes. Since CD69 engagement, together with submitogenic doses of phorbol esters, transduces an activation signal to T lymphocytes, we undertook a comparative study on the stimulation mediated through this receptor on human CD3+ cells cultured under conditions similar to those which occur during exposure to microgravity, i.e. in clinorotation, or at 1 g. During the early hours of activation, increased levels of intracellular calcium and increased mitochondrial membrane potential were detectable in clinorotating as well as in 1 g cells. However, after 48 hours clinorotation, interleukin 2 production by T lymphocytes was significantly reduced and cell proliferation was greatly decreased. By means of a differential proteomics approach on T cells activated in clinorotation or at 1 g for 48 hours, we were able to detect statistically significant quantitative protein alterations. Seven proteins with modified expression values were identified; they are involved in nucleic acids processing, proteasome regulation and cytoskeleton structure. PMID- 15825148 TI - Subcellular proteomic analysis of host-pathogen interactions using human monocytes exposed to Yersinia pestis and Yersinia pseudotuberculosis. AB - Yersinia pestis, the etiological agent of plague, is of concern to human health both from an infectious disease and a biodefense perspective. While Y. pestis and Yersinia pseudotuberculosis share more than 90% DNA homology, they have significantly different clinical manifestations. Plague is often fatal if untreated, yet Y. pseudotuberculosis causes severe intestinal distress but is rarely fatal. A better understanding of host response to these closely related pathogens may help explain the different mechanisms of virulence and pathogenesis that result in such different clinical outcomes. The aim of this study was to characterize host protein expression changes in human monocyte U937 cells after exposure to Y. pestis and Y. pseudotuberculosis. In order to gain global proteomic coverage of host response, proteins from cytoplasmic, nuclear and membrane fractions of host cells were studied by two-dimensional differential gel electrophoresis and relative protein expression differences were quantitated. Differentially expressed proteins, with at least 1.5-fold expression changes and p values of 0.01 or less, were identified by mass spectrometry including matrix assisted laser desorption/ionization-MS or liquid chromatography tandem mass spectrometry. With these criteria, differential expression was detected in 16 human proteins after Y. pestis exposure and 13 human proteins after Y. pseudotuberculosis exposure, of which only two of the differentially expressed proteins identified were shared between the two exposures. Proteins identified in this study are reported to be involved in a wide spectrum of cellular functions and host defense mechanisms including apoptosis, cytoskeletal rearrangement, protein synthesis and degradation, DNA replication and transcription, metabolism, protein folding, and cell signaling. Notably, the differential expression patterns observed can distinguish the two pathogen exposures from each other and from unexposed host cells. The functions of the differentially expressed proteins identified provide insight on the different virulence and pathogenic mechanisms of Y. pestis and Y. pseudotuberculosis. PMID- 15825149 TI - Proteomics of breast cancer: enhanced expression of cytokeratin19 in human epidermal growth factor receptor type 2 positive breast tumors. AB - The human epidermal growth factor receptor, type 2 (HER-2/neu or c-erbB-2) is a 185 kDa transmembrane protein that is phosphorylated upon ligand binding and dimerization with members of the HER/c-erbB family and regulates cell growth and differentiation. Its overexpression is strongly associated with advanced disease, metastasis and poor clinical outcome. To better understand the mechanisms underlying the poor prognosis of breast tumors with HER-2/neu-positive status, parallel proteomic analyses were performed on estrogen receptor-negative and node positive breast tumors with or without overexpression of the HER-2/neu oncogene, using laser capture microdissection and two-dimensional gel electrophoresis. The differentially expressed proteins were identified by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Cytokeratin 19 (CK19), one of the identified proteins, was highly expressed in the HER-2/neu-positive breast tumors when compared with HER-2/neu negative breast tumors. The enhanced overexpression of CK19 in HER-2/neu-positive tumors was further analyzed using semiquantitative reverse-transcription polymerase chain reaction, Western blotting and reverse-phase protein arrays. Immunohistochemical staining of sections from a breast tumor tissue microarray of 97 tumors showed moderate to strong staining against anti-CK19 antibody in 20 (5 with moderate and 15 with strong staining) of the 26 HER-2/neu-positive tumors (76.9%) and in 22 (12 with moderate and 10 with strong staining) of 52 HER-2/neu negative tumors (48%) (p = 0.002). Our results indicate that CK19, an intermediate fragment of the cytoskeleton, and other proteins showing differential expression, are likely to be intricately involved in intra- and intercellular molecular events driving the more aggressive tumor proliferation, invasion and metastasis associated with HER-2/neu-positive tumors. PMID- 15825150 TI - Versatile protein microarray based on carbohydrate-binding modules. AB - Non-DNA microarrays, such as protein, peptide and small molecule microarrays, can potentially revolutionize the high-throughput screening tools currently used in basic and pharmaceutical research. However, fundamental obstacles remain that limit their rapid and widespread implementation as an alternative bioanalytical approach. These include the prerequisite for numerous proteins in active and purified form, ineffectual immobilization strategies and inadequate means for quality control of the considerable numbers of multiple reagents. This study describes a simple yet efficient strategy for the production of non-DNA microarrays, based on the tenacious affinity of a carbohydrate-binding module (CBM) for its three-dimensional substrate, i.e., cellulose. Various microarray formats are described, e.g., conventional and single-chain antibody microarrays and peptide microarrays for serodiagnosis of human immunodeficiency virus patients. CBM-based microarray technology overcomes many of the previous obstacles that have hindered fabrication of non-DNA microarrays and provides a technically simple but effective alternative to conventional microarray technology. PMID- 15825151 TI - Distinct proteome features of plasma microparticles. AB - Plasma microparticles (MPs) are spherical cell membrane fragments derived from either apoptotic or activated cells. Characterized by a rich phospholipid moiety and many protein constituents, MPs normally circulate in the blood and contribute to numerous physiological processes. In disease states, MPs derived from the injured organ likely contain valuable markers for determining the site, type, and extent of disease pathology. However, the basic protein characteristics of plasma MPs have yet to be described. In this study, MPs from a pooled plasma sample derived from 16 healthy donors, all of group A blood type, were prepared by ultracentrifugation. Flow cytometry confirmed that a majority of these MPs are smaller than 1 microm. Factor Xa generation assay revealed the presence of tissue factor activity in these MPs, confirming MPs' role in initiating blood coagulation. The MP proteome was analyzed by two-dimensional (2-D) gel electrophoresis performed in triplicate, and compared with a 2-D gel of pooled whole plasma and blood platelets. Overall, plasma MPs displayed distinct protein features and a greater number of protein spots (1021-1055) than that detected in whole plasma (331-370). Protein spots expressed in high abundance in the MP proteome were then excised and submitted for protein identity determination. This process provided protein identification for 169 protein spots and reported their relative protein quantities within the MP proteome. These 169 protein spots represented 83 different proteins and their respective isoforms. Thirty of these proteins have never before been reported in previous proteome analyses of human plasma. These results provide unprecedented information on the MP proteome and create a basis for future studies to understand MP biology and pathophysiology. PMID- 15825152 TI - Proteomic changes associated with inactivation of the Candida glabrata ACE2 virulence-moderating gene. AB - Inactivation of the gene encoding the transcriptional activator Ace2 in the fungal pathogen Candida glabrata results in an almost 200-fold increase in virulence characterised by acute mortality and a massive over-stimulation of the pro-inflammatory arm of the innate immune system. In this study we have adopted a proteomics approach to identify cellular functions regulated by C. glabrata Ace2 that might contribute to this increase in virulence. A two-dimensional polyacrylamide gel electrophoresis map of the C. glabrata proteome was constructed. We identified a total of 123 proteins, 61 of which displayed reproducible and statistically significant alterations in their levels following inactivation of ACE2. Of these, the levels of 32 proteins were elevated, and 29 were reduced in ace2 cells. These data show that Ace2 influences metabolism, protein synthesis, folding and targeting, and aspects of cell growth and polarisation. Some of these functions are likely to contribute to the effects of Ace2 upon the virulence of C. glabrata. PMID- 15825153 TI - Chronic pudendal nerve neuromodulation in women with idiopathic refractory detrusor overactivity incontinence: results of a pilot study with a novel minimally invasive implantable mini-stimulator. AB - AIMS: To assess the results of chronic pudendal nerve neuromodulation with a novel technique in a pilot study on women with idiopathic refractory detrusor overactivity incontinence. MATERIALS AND METHODS: A percutaneous screening test (PST) was performed in patients with urodynamically demonstrated detrusor overactivity incontinence. Such a test includes the performance of a cystometrogram without and with percutaneous pudendal nerve stimulation and is considered positive if stimulation results in a more than 50% increase in the bladder volume at the first involuntary detrusor contraction or the maximum cystometric capacity. Patients with a positive PST qualified for the implantation of a mini-neurostimulator with an integrated electrode, a so-called bion(R), adjacent to the pudendal nerve at Alcock's Canal. Five-day voiding-incontinence diaries were the main tool for the evaluation of therapy. RESULTS: A PST was performed in 14 women. Six patients responded positively and received a bion. The degree of incontinence decreased significantly in this group, which also included patients in whom sacral neuromodulation had failed. There were no severe adverse events. CONCLUSIONS: Chronic pudendal nerve stimulation may reduce the degree of detrusor overactivity incontinence, even in patients in whom sacral neuromodulation fails. Bion therapy is relatively simple and minimally invasive and is well tolerated by the patient. PMID- 15825154 TI - Mechanistic studies on the tyrosinase-catalyzed formation of the anachelin chromophore. AB - The complex secondary metabolite anachelin, isolated from the freshwater cyanobacterium Anabaena cylindrica, is believed to act as siderophore, facilitating iron uptake. Its structure is characterized by a fascinating blend of polyketide, peptide, and alkaloid fragments. In particular, the tetrahydroquinolinium-derived chromophore is unique among natural products, and its biosynthesis is unknown. We propose a hypothesis for the biogenesis of the anachelin chromophore starting from a C-terminally bound L-Tyr residue. It is proposed that this amino acid is reductively aminated, methylated, and hydroxylated. Oxidation of this catechol diamine substrate by a tyrosinase would lead to an o-quinone, which would react by intramolecular aza-annulation and tautomerization to give the anachelin chromophore. In order to evaluate this hypothesis, a model substrate related to the proposed biogenetic precursor was prepared. It was shown that the enzyme tyrosinase is able to transform this substrate into an anachelin chromophore derivative, which corroborates the biogenetic hypothesis. In order to gain further insight into the mechanism of this transformation, we performed spectrophotometric reaction monitoring, allowing the formation of the expected product to be observed. In addition, a rise in absorption at around 250 nm might be due to the presence of a spiro five membered ring intermediate resulting from an alternative 1,4-addition to the o quinone. Lastly, we were able to show that the action of tyrosinase on this substrate follows Michaelis-Menten kinetics (k(cat)=123 s(-1) and K(m)=8.66 mM). Interestingly, the catalytic efficiency is decreased only by a factor of 30 relative to the natural substrate L-DOPA. PMID- 15825155 TI - Pathology of radiation and anthracycline cardiotoxicity. AB - Radiation-induced heart disease (RIHD) and anthracycline cardiotoxicity are two patterns of cardiac dysfunction caused by therapeutic interventions to treat malignancies. They occur in both the pediatric and adult populations and there is evidence to suggest that pediatric patients are at greater risk. This is due in part to the longer survival rates but also reflects increased susceptibility to the attendant complications caused by both therapies. Radiation can cause injury to all the components of the heart, including the vasculature, while anthracycline toxicity is generally limited to the myocardium. PMID- 15825157 TI - Prevention of coronary artery disease in cancer patients. AB - Improvements in cancer treatment are increasing survival rates, placing cancer survivors at increased risk of coronary artery disease (CAD). Thoracic radiation as cancer therapy can accelerate the atherosclerotic process, and some older studies suggest that mediastinal radiation can increase the CAD risk two to fivefold. The management of serum lipids using diet and drugs is the cornerstone of efforts designed to reduce CAD events. The present article reviews the association between radiation and coronary disease and provides guidelines for the prevention of atherosclerotic CAD in this patient group and in the general population of cancer survivors. PMID- 15825158 TI - Reactions of ion-pair intermediates of solvolysis. AB - The detection during solvolysis of competing signature reactions such as the racemization of a chiral substrate, or the scrambling of oxygen isotopes at the leaving group serves as evidence for a stepwise mechanism that proceeds through a reversibly formed ion-pair intermediate. We have examined these signature reactions and determined the relative rates of competing solvolysis, isomerization, and racemization reactions of neutral ring-substituted 1 phenylethyl derivatives. These data were used to define the relative rate constants for partitioning of carbocation-anion pairs between dynamic processes such as reorganization of the ion pair within a solvent cage, and migration of the leaving group anion from the "top" to the "bottom" face of the planar carbocation. We present here a well-defined picture of the dynamics of these reactions in the mostly aqueous solvent of 50/50 (v/v) trifluoroethanol/water. A more complicated problem is the description of the changes in the reaction energy profiles for solvolysis, isomerization, and racemization reactions that occur as one destabilizes the carbocation intermediate of the stepwise reaction. We have examined the signature isomerization and racemization reactions of a ring substituted 1-phenylethyl derivative that reacts by a "borderline" mechanism. We suggest that the signature isomerization reaction may in fact follow a concerted mechanism that bypasses the putative carbocation-anion pair intermediate. PMID- 15825159 TI - Spatial distributions of desorbing products in basic catalytic processes and surface nano-structures. AB - Reviews of recent progress in angle-resolved measurements of desorbing surface reaction products are discussed. The angular and velocity distributions of desorbing products deliver information about the reaction site as well as the reaction mechanism when the products are repulsively desorbed. These distribution measurements can yield symmetry and orientation information of the reaction site for associative processes whereas, in dissociative desorption, the collimation of fragment desorption is related to the orientation of the intermediate species immediately before dissociation. These different collimations provide information on desorption steps whenever any step becomes rate determining. PMID- 15825160 TI - Retromolar trigone squamous cell carcinoma treated with radiotherapy alone or combined with surgery. AB - BACKGROUND: Treatment outcomes were analyzed for retromolar trigone squamous cell carcinoma. METHODS: Between June 1966 and August 2003, 99 patients were treated with radiotherapy alone (35 patients) or radiotherapy combined with surgery (64 patients). Followup ranged from 0.2 to 23.8 years (median, 3.3 yrs). All living patients had followup for at least 1 year. RESULTS: The 5-year local-regional control rates after definitive radiotherapy versus surgery and radiotherapy were as follows: Stages I-III, 51% and 87%; Stage IV, 42% and 62%; and overall, 48% and 71%, respectively. The 5-year cause-specific survival rates after definitive radiotherapy compared with surgery and radiotherapy were as follows: Stages I III, 56% and 83%; Stage IV, 50% and 61%; and overall, 52% and 69%, respectively. Multivariate analyses revealed that the likelihood of cure was better with surgery and radiotherapy compared with definitive radiotherapy. CONCLUSIONS: The likelihood of cure after treatment for retromolar trigone squamous cell carcinoma was influenced by the extent of disease and treatment. Patients treated with surgery and radiotherapy had a better outcome than those treated with radiotherapy alone. PMID- 15825161 TI - The efficacy of epirubicin, cisplatin, uracil/tegafur, and leucovorin in patients with advanced biliary tract carcinoma. AB - BACKGROUND: Advanced biliary tract carcinoma is among the most prevalent fatal diseases in Korea. However, to our knowledge, to date no effective therapeutic modality has been shown to prolong the survival of patients in the inoperable stages of this disease. METHODS: This Phase II study was conducted to determine the efficacy and toxicity of a combined regimen of epirubicin, cisplatin, and uracil/tegafur (UFT) modulated by leucovorin in patients with advanced or recurrent biliary tract carcinoma. RESULTS: Eleven of 40 patients (27.5%) had gallbladder carcinoma, and the remaining patients had tumors arising from other sites in the biliary tract. All patients were treated with intravenous epirubicin (50 mg/m(2) on Day 1), intravenous cisplatin (60 mg/m(2) on Day 1), oral UFT (300 mg/m(2) per day on Days 1-21), and oral leucovorin (75 mg per day on Days 1-21). Nine patients exhibited a partial response, representing 22.5% of the possible response rate (95% confidence interval [95% CI], 12.8-32.2%) based on an intention-to-treat analysis. The median survival was 34 weeks (95% CI, 20-48 weeks), and the median time to disease progression was 16 weeks (95% CI, 7-25 weeks). Neutropenia and thrombocytopenia comprised dose-limiting toxicity conditions. CONCLUSIONS: The combination of epirubicin, cisplatin, and UFT modulated by leucovorin was active marginally in patients with advanced biliary tract carcinoma and was capable of stabilizing the disease effectively. Because it was a safe and convenient treatment modality, it may be used in outpatient care with only minor toxicity in patients with advanced malignancies of the biliary tract. PMID- 15825162 TI - Clinical role of tissue and serum levels of SCCA antigen in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most frequent cancer in the world and a common occurrence in patients with liver cirrhosis in western and North American countries. Ultrasound screening is a powerful technique for HCC diagnosis, whereas the only available serologic test, alpha-fetoprotein, has poor reliability. It has been reported that the squamous cell carcinoma antigen (SCCA) is overexpressed in HCC tissue. In our study, the expression of SCCA was investigated in tumoral and peritumoral tissues and in the serum of 52 HCC patients, as well as in the serum of 48 cirrhotic patients. The results show that SCCA expression is much stronger in the tumoral than in the peritumoral tissue of HCC. Moreover, it is also evident in metastatic nodules present in the peritumoral tissue. SCCA serum levels were significantly higher in HCC samples than in cirrhotic samples. However, no correlation was found between SCCA expression and the HCC histologic degree, nor did SCCA expression correlate with tumor size, presence of metastasis or clinical outcome. In conclusion, in HCC patients, the SCCA antigen could represent a useful marker for the detection of micro-metastasis in the tissues and for large-scale screening of serum in patients at risk. PMID- 15825163 TI - COX-2 inhibitors suppress integrin alpha5 expression in human lung carcinoma cells through activation of Erk: involvement of Sp1 and AP-1 sites. AB - Tumor cell expression of COX-2 has been implicated in the progression of murine and human lung cancer. Inhibition of COX-2 by nonsteroidal antiinflammatory drugs reduces the risk of cancer development in humans and suppresses tumor growth in animal models. However, the underlying mechanisms for this beneficial effect are not fully understood. Here we explore the potential link between the anticancer effects of COX-2 inhibitors and the expression of the integrin alpha5beta1. Expression of this integrin in carcinoma cells is associated with invasiveness and malignant progression. This, together with our studies showing that fibronectin, the ligand of alpha5beta1, stimulates the growth of human lung carcinoma cells, and that this effect is mediated through alpha5beta1-dependent signals, has prompted us to examine the effects of COX-2 inhibitors on alpha5beta1 expression in human non small cell lung carcinoma (NSCLC) cells. We found that the selective COX-2 inhibitors NS398 and Nimesulide decreased mRNA expression and protein production of the integrin alpha5 subunit. This effect was associated with inhibition of NSCLC cell adhesion to fibronectin. The COX-2 inhibitors triggered the phosphorylation of extracellular signal-regulated kinase (Erk) in a time-dependent manner, and the inhibitor of Mek-1/Erk PD98095 prevented their inhibitory effects on integrin alpha5 expression. Transient transfection assays showed that the COX-2 inhibitors affected integrin alpha5 gene transcription by acting between -92 to -41 bp of the human integrin alpha5 gene promoter. Gel mobility shift assays showed that the COX-2 inhibitors increased Sp1 DNA binding, but decreased that of AP-1. These effects were accompanied by an increase in Sp1 protein and a decrease in c-Jun protein expression, as well as inhibition of SAPK/JNK phosphorylation. The Sp1 inhibitor, Mithramycin A, also blocked the inhibitory effect of the COX-2 inhibitors on alpha5 expression and promoter activity. Overall, these findings suggest that COX 2 inhibitors suppress alpha5beta1 integrin expression in NSCLC through effects on integrin alpha5 gene transcription mediated by Erk activation, increased Sp1, decreased AP-1 DNA binding and inactivation of SAPK/JNK signals. Our observations unveil a new mechanism of action against NSCLC for COX-2 inhibitors that relates to regulation of integrin alpha5 gene expression and, consequently, recognition of extracellular matrices (i.e., fibronectin) by tumor cells. (c) 2005 Wiley Liss, Inc. PMID- 15825164 TI - Preparation and characterization of calibration beads for sorting cells expressing a beta-lactamase gene reporter. AB - BACKGROUND: Modern drug discovery has been based on high-throughput screening using whole-cell assays. A prominent role has been assigned to the reporter gene technology based on a beta-lactamase and the fluorogenic substrate CCF2. Successful application of this technology requires fluorescence-activated cell sorting. We describe the preparation and characterization of calibration beads for sorting cells expressing the beta-lactamase gene using the CCF2 substrate. METHODS: To model Forster resonance energy transfer (FRET) between the coumarin donor and the fluorescein acceptor of the CCF2 reporting dye, we used activated polystyrene beads with primary amino groups. Donor and acceptor fluorophores were attached to the beads at different ratios via succinimidyl esters. The beads were characterized with a fluorescence plate reader and a flow cytometer. RESULTS: We prepared polystyrene beads with five different ratios of donor and acceptor fluorophores and beads that carried a donor or a receptor fluorophore alone. Fluorescence measurements demonstrated that the prepared beads well represent the FRET of CCF2 substrate. CONCLUSION: We have demonstrated that the prepared beads can be successfully used for the setup of fluorescence-activated cell sorting to sort cells with CCF2 reporter substrate and the beta-lactamase reporter gene. PMID- 15825165 TI - Persistent oligoclonal CD4dimCD8+T cells in peripheral blood. AB - BACKGROUND: Routine CD4/CD8 T-cell phenotyping may shows a small fraction of CD4dimCD8+ T cells with a homogeneous appearance as described for lymphoproliferative syndromes or chronic infections. The aim of this study was to elucidate the significance of CD4dimCD8+ T cells and their degree of diversity. METHODS: Phenotyping was performed in 272 samples from healthy donors, elderly patients, and immunocompromised (human immunodeficiency virus or renal transplantation) patients. RESULTS: The CD4dimCD8+ T cells had decreased fluorescence intensity for CD4 but not for CD8. The frequency of patients with CD4dimCD8+ T cells (>20 cells/microl; 10.3% of patients with human immunodeficiency virus and 7.7% with renal transplantation) was not significantly different when compared with healthy donors (9.7%). The CD4dimCD8+ T cells did not express the activation marker CD69. The CD8 of CD4dimCD8+ T cells expressed the heterodimeric (beta) isoform. In 13 of 26 samples, the apparently highly homogeneous CD4dimCD8+ T cells expressed one predominant T-cell receptor Vbeta clonotype. These predominant clonotypes were widely distributed among patients: Vbeta 5.2, 17, 2, 3, 5.1, 13.1, 14, and 20. CONCLUSIONS: Whether these findings demonstrate an oligoclonal reaction to chronic inflammation or an emerging lymphoproliferative disorder must be elucidated in a long-term longitudinal study. By analogy to monoclonal gammopathy, we propose to name this phenomenon "oligoclonal clonopathy of undetermined significance." PMID- 15825166 TI - Effects of loss of p53 and p16 function on life span and survival of human urothelial cells. AB - Human urothelial cell carcinoma evolves via the accumulation of numerous genetic alterations, with loss of p53 and p16 function representing important stages in the development of superficial lesions and their progression to malignant disease. To investigate the effects of disabling either or both proteins in otherwise normal human urothelial cells, we performed retroviral transductions with a dominant negative p53 miniprotein and/or mutant cyclin-dependent kinase 4 (CDK4R24C) in 3 independent cell lines. Although cells with disabled p53 function showed a higher proliferation rate, inactivation of neither p53 nor p16 function resulted in any extension of life span and the double-transductants failed to flourish, demonstrating that further genetic alterations are required to attain an immortalised phenotype. However, CDK4R24C transductants showed a marked increase in apoptotic susceptibility to membrane-presented CD40 ligand, being intermediate between normal cells (nonsusceptible) and transformed cells (highly susceptible). By contrast, loss of p53 function alone only slightly increased the apoptotic susceptibility of urothelial cells. These results demonstrate that loss of p16 function, while insufficient to immortalise urothelial cells, nevertheless renders the cells more vulnerable to apoptosis induced by CD40 ligation. PMID- 15825167 TI - Intratumoral injection of immature dendritic cells enhances antitumor effect of hyperthermia using magnetic nanoparticles. AB - Dendritic cells (DCs) are potent antigen-presenting cells that play a pivotal role in regulating immune responses in cancer and have recently been shown to be activated by heat shock proteins (HSPs). We previously reported that HSP70 expression after hyperthermia induces antitumor immunity. Our hyperthermia system using magnetite cationic liposomes (MCLs) induced necrotic cell death that was correlated with HSP70 release. In the present study, we investigated the therapeutic effects of DC therapy combined with MCL-induced hyperthermia on mouse melanoma. In an in vitro study, when immature DCs were pulsed with mouse B16 melanoma cells heated at 43 degrees C, major histocompatibility complex (MHC) class I/II, costimulatory molecules CD80/CD86 and CCR7 in the DCs were upregulated, thus resulting in DC maturation. C57BL/6 mice bearing a melanoma nodule were subjected to combination therapy using hyperthermia and DC immunotherapy in vivo by means of tumor-specific hyperthermia using MCLs and directly injected immature DCs. Mice were divided into 4 groups: group I (control), group II (hyperthermia), group III (DC therapy) and group IV (hyperthermia + DC therapy). Complete regression of tumors was observed in 60% of mice in group IV, while no tumor regression was seen among mice in the other groups. Increased cytotoxic T lymphocyte (CTL) and natural killer (NK) activity was observed on in vitro cytotoxicity assay using splenocytes in the cured mice treated with combination therapy, and the cured mice rejected a second challenge of B16 melanoma cells. This study has important implications for the application of MCL-induced hyperthermia plus DC therapy in patients with advanced malignancies as a novel cancer therapy. PMID- 15825169 TI - Selective methodologies for the synthesis of biologically active piperidinic compounds. AB - The synthesis of optically active substituted piperidines has been achieved by using four different methodologies. The first one is an intramolecular nucleophilic displacement of activated alcohol moieties that was used to build up the piperidine ring of (-)-prosophylline and (-)-slaframine, and the second one is a ring-closing metathesis of unsaturated amines which was employed in the synthesis of (+)-sedamine and 4a,5-dihydrostreptazoline. The third methodology is the alpha-functionalization of N-Boc piperidines which was particularly useful in the synthesis of argatroban, and the fourth one is a ring expansion of prolinols to 3-chloropiperidines or 3-hydroxypiperidines which was utilized to synthesize ( )-paroxetine, (-)-pseudoconhydrine, the piperidine ring of (-)-velbanamine and (+)-zamifenacin. PMID- 15825168 TI - Lack of guanylyl cyclase C, the receptor for Escherichia coli heat-stable enterotoxin, results in reduced polyp formation and increased apoptosis in the multiple intestinal neoplasia (Min) mouse model. AB - Guanylyl cyclase C (GC-C), a transmembrane receptor for bacterial heat-stable enterotoxin and the mammalian peptides guanylin and uroguanylin, mediates intestinal ion secretion and affects intestinal cell growth via cyclic GMP signaling. In intestinal tumors, GC-C expression is maintained while guanylin and uroguanylin expression is lost, suggesting a role for GC-C activation in tumor formation or growth. We show by in situ hybridization that GC-C expression is retained in adenomas from multiple intestinal neoplasia (Apc(Min/+)) mice. In order to determine the in vivo role of GC-C in intestinal tumorigenesis, we generated Apc(Min/+) mice homozygous for a targeted deletion of the gene encoding GC-C and hypothesized that these mice would have increased tumor multiplicity and size compared to wild-type Apc(Min/+) mice on the same genetic background. In contrast, the absence of GC-C resulted in a reduction of median polyp number by 55%. There was no change in the median diameter of polyps, suggesting no effect on tumor growth. Somatic loss of the wild-type Apc allele, an initiating event in intestinal tumorigenesis, also occurred in polyps from GC-C-deficient Apc(Min/+) mice. We have found increased levels of apoptosis as well as increased caspase-3 and caspase-7 gene expression in the intestines of GC-C-deficient Apc(Min/+) mice compared with Apc(Min/+) mice. We propose that these alterations are a possible compensatory mechanism by which loss of GC-C signaling also affects tumorigenesis. PMID- 15825170 TI - Dietary patterns and risk of prostate cancer in Ontario, Canada. AB - Dietary patterns reflect combinations of dietary exposures, and here we examine these in relation to prostate cancer risk. In a case-control study, 80 incident primary prostate cancer cases and 334 urology clinic controls were enrolled from 1997 through 1999 in Kingston, Ontario, Canada. Food-frequency questionnaires were completed prior to diagnosis and assessed intake in the 1-year period 2-3 years prior to enrollment. Among controls, dietary intake was used in principal components analyses to identify patterns that were then evaluated with all subjects in relation to prostate cancer risk using unconditional logistic regression, controlling for age. Four dietary patterns were identified: Healthy Living, Traditional Western, Processed and Beverages. Increased prostate cancer risk is apparent in relation to the Processed pattern, composed of processed meats, red meats, organ meats, refined grains, white bread, onions and tomatoes, vegetable oil and juice, soft drinks and bottled water. The OR for the highest tertile compared to baseline is 2.75 (95% CI 1.40-5.39), with a dose-response pattern (trend test p < 0.0035). Our results suggest that a dietary pattern including refined grain products, processed meats and red and organ meats contributes to increased prostate cancer risk. Since dietary information was collected before subjects knew their diagnosis, recall bias was avoided. PMID- 15825171 TI - Novel real-time PCR assay using a universal molecular marker for diagnosis of hematologic cancers. PMID- 15825172 TI - TX-1877, a bifunctional hypoxic cell radiosensitizer, enhances anticancer host response: immune cell migration and nitric oxide production. AB - We investigated in the current study the effect of TX-1877, a bifunctional hypoxic cell radiosensitizer, in augmenting anticancer host response. In the syngeneic squamous cell carcinoma-bearing mouse model, a single administration of TX-1877 significantly inhibited the primary tumor growth as well as lung metastasis. TX-1877 administration resulted in a significant infiltration of immune cells, such as CD4+T, CD8+T cells, macrophages and dendritic cells (DCs), and an increased expression of chemokines for cytotoxic T lymphocytes (CTLs), helper T-cell 1 (Th1) cells, monocytes/macrophages and DCs, in tumor tissues. Nitric oxide (NO) production and the expression of inducible NO synthase (iNOS) and interferon-gamma, a major Th1 cytokine that plays a major role in anticancer immunity, were also enhanced. Furthermore, neutralization of NO by N-monomethyl-L arginine acetate resulted in a marked inhibition of the antitumor effect of TX 1877. In tumor-draining lymph nodes, MHC class I-restricted CD8+ memory CTLs specific for inoculated cancer cells were induced by TX-1877. In in vitro experiments, TX-1877 induced chemokines and iNOS/NO in several types of culture cells. These findings strongly suggested that TX-1877 induces migration of CD8+CTLs, CD4+Th1 cells, macrophage/monocytes and dendritic cells into the tumor site, and that this migration is mediated by chemokine induction. In addition, it was suggested that NO produced by several types of cells stimulated by TX-1877 in the tumor sites plays a major role in the anticancer effect of TX-1877. TX-1877 was thus shown to be an effective immunopotentiator as well as a hypoxic cell radiosensitizer. PMID- 15825173 TI - Absence of the ERBB2 kinase domain mutation in lung adenocarcinomas in Korean patients. PMID- 15825176 TI - Polarized light out-coupling from lightguides for LCDs. AB - New designs of lightguide systems, which emit linear polarized light with a high efficiency for transmissive and transflective LCD applications, are presented. These systems are equipped with nano- and/or micro-structured films or coatings, which emit highly collimated or diffuse linearly polarized light with a high efficiency. The films are based on polarization-selective scattering, reflection, or diffraction of light and their properties can be tuned to a large extent dependent on their envisioned application. For instance, edge-lit lightguide systems are discussed, which combine a range of desirable features such as a high transparency in direct view, a direct emission of light at normal angles to the plane of the lightguide, and a purely unidirectional out-coupling of light towards the LCD-side. PMID- 15825175 TI - Aberrant methylation of secreted frizzled-related protein genes in esophageal adenocarcinoma and Barrett's esophagus. AB - Hypermethylation of secreted frizzled-related proteins (SFRP) genes frequently occurs with several cancers but has not been studied in esophageal adenocarcinoma or its precursor-Barrett's esophagus. To explore the role of SFRP methylation in the neoplastic progression of Barrett's esophagus and to evaluate methylated SFRP genes as biomarkers for Barrett's esophagus and cancer, methylation of SFRP genes was determined in esophageal adenocarcinomas, Barrett's esophagus and normal epithelia using methylation-specific PCR. Protein expression of SFRP genes was then assessed in these tissues by immunohistochemistry. The mRNA expression of SFRP genes was quantified by real-time reverse-transcription PCR in esophageal adenocarcinoma cell lines with and without demethylation by 5-aza-2'deoxycytidine and inhibition of deacetylation by trichostatin A treatment. Hypermethylation of SFRP1, 2, 4 and 5 was detected in 93%, 83%, 73% and 85% of 40 cancers; 81%, 89%, 78% and 73% of 37 Barrett's epithelia; 25%, 64%, 32% and 21% of 28 adjacent normal epithelia from Barrett's patients; and 10%, 67%, 0% and 13% of 30 normal esophagogastric epithelia from healthy individuals, respectively (p < 0.001 for SFRP1, 4 and 5; p < 0.05 for SFRP2). Protein expression of SFRP1, 2 and 4 was downregulated in 87%, 67% and 90% of cancers, and expression correlated inversely with grade and stage of cancers and with grade of dysplasia. Expression of SFRP2 and SFRP4 proteins was lower in cancers with corresponding gene methylation (p < 0.05). Demethylation treatment effectively re-expressed SFRP mRNA in cancer cell lines. Thus, hypermethylation of SFRP genes is a common early event in the evolution of esophageal adenocarcinoma, and methylation of SFRP1, 4 and 5 might serve as biomarkers for Barrett's neoplasia. Aberrant promoter methylation appears to functionally silence SFRP gene expression in esophageal adenocarcinoma. PMID- 15825177 TI - Matrix metalloproteinases activity and stromal-epithelial interactions. PMID- 15825178 TI - Genomic gains on chromosome 1q in retinoblastoma: consequences on gene expression and association with clinical manifestation. AB - Many retinoblastomas (Rbs) show genomic alterations in addition to mutational loss of both normal RB1 alleles. The most frequent of these changes are gains on chromosomes 1q and 6p and losses on 16q. To identify the genes targeted by gains on chromosome 1q, we used quantitative-multiplex PCR to determine DNA copy number changes in 76 primary tumors and 6 Rb cell lines. In addition, in 21 of these tumors, gene expression was analyzed by cDNA microarray hybridization. Increased copy numbers of loci on chromosome 1q were present in 34 (45%) primary tumors and in all 6 cell lines. Two regions of gain emerged, one in 1q32 and another in 1q21. Tumors with 1q gains showed higher RNA expression of several genes in these 2 regions. The clinical manifestation of tumors with and without gains was similar with regard to many aspects, including size, necrosis and calcification. However, the distribution of age at diagnosis was remarkably distinct, with earlier diagnosis in tumors without gains. This suggests that these tumors either are initiated earlier or grow faster than tumors with gains. This association with clinical manifestation indicates that gains on 1q are significant for the biology of Rb. The genes on 1q with copy number gains and overexpression are candidates that need to be tested for their individual contribution to the progression of Rb. PMID- 15825179 TI - SPOC1, a novel PHD-finger protein: association with residual disease and survival in ovarian cancer. AB - We report the identification of a novel human gene (SPOC1) which encodes a protein with a PHD-finger domain. The gene is located in chromosomal region 1p36.23, a region implicated in tumor development and progression. RNA in situ hybridization experiments showed strong SPOC1 expression in some rapidly proliferating cell types, such as spermatogonia, but not in nonproliferating mature spermatocytes. In addition, high SPOC1 mRNA expression was observed in several ovarian cancer cell lines. This prompted us to systematically examine SPOC1 expression in ovarian cancer in relation to prognosis. SPOC1 mRNA expression was quantified in tumor tissue of 103 patients with epithelial ovarian cancer. Interestingly, SPOC1 was associated with residual disease, whereby patients with unresectable tumors showed higher levels compared to patients without residual tumor tissue after surgery (p = 0.029). The univariable proportional hazards model showed an association between SPOC1 expression and survival (p = 0.043, relative risk = 1.535). Median survival time was 1,596 days for patients with low SPOC1 expression vs. only 347 days for patients with high expression, using Kaplan-Meier analysis. However, SPOC1 was not associated with survival when multivariable analysis was adjusted for residual disease. This can be explained by the correlation between residual disease and SPOC1 expression. In conclusion, SPOC1 is a novel PHD-finger protein showing strong expression in spermatogonia and ovarian cancer cells. SPOC1 overexpression was associated with unresectable carcinomas and shorter survival in ovarian cancer. PMID- 15825180 TI - Selecting the right fluorophores and flow cytometer for fluorescence resonance energy transfer measurements. AB - BACKGROUND: Fluorescence resonance energy transfer applied in flow cytometry (FCET) is an excellent tool for determining supramolecular organization of biomolecules at the cell surface or inside the cell. Availability of new fluorophores and cytometers requires the establishment of fluorophore dye pairs most suitable for FCET measurements. METHODS: A gastric tumor cell line (N87) was labeled for major histocompatibility complex class I heavy chain and beta2 microglobulin with antibodies conjugated with fluorescein- and indocarbocyanine like fluorophores and analyzed in FCET measurements on a cell-by-cell basis using three flow cytometers: FACSCalibur, FACSDiVa, and FACSArray. RESULTS: Normalized fluorescence intensity values were measured and normalized energy transfer efficiencies, spectral overlap integrals, and crucial dye- and instrument dependent parameters were calculated for all matching pairs of seven fluorophores on the three commercial cytometers. The most crucial parameter in determining the applicability of the donor-acceptor pairs was the normalized fluorescence intensity and the least important one was the spectral overlap. CONCLUSIONS: On the basis of available laser lines, the optimal dye pair for all three cytometers is the Alexa546-Alexa647 pair, which produces high energy transfer efficiency values and has the best spectral characteristics with regard to laser excitation, detection of emission, and spectral overlap. PMID- 15825181 TI - Impedance spectroscopy flow cytometry: on-chip label-free cell differentiation. AB - BACKGROUND: The microfabricated impedance spectroscopy flow cytometer used in this study permits rapid dielectric characterization of a cell population with a simple microfluidic channel. Impedance measurements over a wide frequency range provide information on cell size, membrane capacitance, and cytoplasm conductivity as a function of frequency. The amplitude, opacity, and phase information can be used for discrimination between different cell populations without the use of cell markers. METHODS: Polystyrene beads, red blood cells (RBCs), ghosts, and RBCs fixed in glutaraldehyde were passed through a microfabricated flow cytometer and measured individually by using two simultaneously applied discrete frequencies. The cells were characterized at 1,000 per minute in the frequency range of 350 kHz to 20 MHz. RESULTS: Cell size was easily measured with submicron accuracy. Polystyrene beads and RBCs were differentiated using opacity. RBCs and ghosts were differentiated using phase information, whereas RBCs and fixed RBCs were differentiated using opacity. RBCs fixed using increasing concentrations of glutaraldehyde showed increasing opacity. This increased opacity was linked to decreased cytoplasm conductivity and decreased membrane capacitance, both resulting from protein cross-linking. CONCLUSIONS: This work presents label-free differentiation of cells in an on-chip flow cytometer based on impedance spectroscopy, which will be a powerful tool for cell characterization. PMID- 15825182 TI - Functional analysis of p53 gene and the prognostic impact of dominant-negative p53 mutation in endometrial cancer. AB - In addition to the loss of function, mutant p53 can possess a dominant-negative effect on wild-type p53 and may also exert gain-of-function activity. It is not clear whether the functional status of p53 mutation contributes to differences in outcome in endometrial cancer. We collected a total of 92 RNA samples of high quality from endometrial cancer tissues, and the samples were subjected to yeast functional assay and sequencing for p53 mutations. The detected mutant p53 genes were further investigated for their dominant-negative activity using a yeast based transdominance assay. p53 mutation was found in 24 out of 92 (26.1%) tumors, of which 10 exhibited no dominant-negative activity (recessive mutation) and 14 showed dominant-negative activity. Dominant-negative p53 mutation was related to advanced stages (p = 0.01), nonendometrioid type tumors (p = 0.01) and grade 3 tumors (p = 0.04). The patients with dominant-negative mutation had significantly shorter survival than patients with no mutation (p < 0.0001) and those with a recessive mutation (p = 0.01) in the p53 gene. No difference in survival was found between the patients with tumors harboring a recessive p53 mutation and those with tumors harboring a wild-type p53. Multivariate analysis revealed that dominant-negative p53 mutation (p = 0.019), FIGO stage (p = 0.0037) and histologic subtype (p = 0.014) were independently related to patient survival. Dominant-negative p53 mutation was the most important prognostic factor for stage III/IV endometrial cancer (p = 0.0023). In conclusion, dominant negative p53 mutation is often found in advanced stages and aggressive histologic subtypes of endometrial cancer and it is a strong predictor of survival of patients with advanced endometrial cancer. To elucidate further the role of p53 mutation in endometrial cancer, it is necessary to investigate gain-of-function activity involving dominant-negative p53 mutant proteins. PMID- 15825183 TI - Flow cytometric quantitative determination of ingestion by phagocytes needs the distinguishing of overlapping populations of binding and ingesting cells. AB - BACKGROUND: The use of flow cytometry with fluorescently labeled particles provides the means to examine quantitatively the phagocytotic capacity of an individual phagocyte. This report describes an improved flow cytometric method of analysis for kinetic measurement of phagocytosis of fluorescein isothiocyanate (FITC)-labeled zymosan particles by human leukocytes. METHODS: FITC-labeled zymosan was incubated with leukocyte suspension, and at selected time intervals fluorescence positive neutrophils were divided by phagocytotic gates into three subpopulations: neutrophils that were neither binding nor ingesting particles, neutrophils that were only binding particles (binding cells), and neutrophils that were binding and ingesting particles (ingesting cells). For the distinction between internalized and surface-bound FITC-labeled zymosan, trypan blue (1.2 mg/ml) was used to quench surface-bound fluorescence. RESULTS: The technical challenges related to settings of phagocytotic gates and derivation of phagocytotic equations were presented. From 28 control samples, numerical values of mean fluorescence intensities and percentages of phagocytotic subpopulations inside phagocytotic gates before and after quenching were inserted into phagocytotic equations and corrected phagocytotic parameters were calculated. Calculated parameters were surprisingly constant across individuals. CONCLUSIONS: Essential elements of the present method appeared to be partial quenching of extracellular fluorescence with trypan blue and distinguishing between overlapping populations of binding and ingesting cells. Corrections using derived phagocytotic equations proved necessary for accurate kinetic phagocytotic measurements. Corrections were less necessary when the ingestion process was finished. PMID- 15825184 TI - Areca (betel) nut extract activates mitogen-activated protein kinases and NF kappaB in oral keratinocytes. AB - Areca (betel) was recently proved a carcinogenic substance by the International Agency for Research on Cancer. However, the signaling impact of areca in oral keratinocyte is still obscure. Mitogen-activated protein kinase superfamilies, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinases (JNK) and p38, together with transcription factor NF-kappaB, are important signaling elements. We examined the activation of these signaling pathways in OECM-1 and SAS oral keratinocytes, treated with ripe areca nut extract (ANE). In both cells, a rapid increase in JNK1 activity at 0.5 hr was noted following treatment of ANE. ERK was profoundly activated during 0.5-2 hr in OECM-1 cells. Contrasting p38 activity was noted in these 2 cells. In both cells, ANE also activated NF-kappaB pathway in a biphasic manner, particularly for SAS cells. NF kappaB was activated by approximately 2- to 4-fold at 0.5-1 hr and a plateau or slight decrease of activity existed between 1 and 6 hr. Later, another higher episode of NF-kappaB activity was raised. This was accompanied with the rapid degradation in cytosolic IkappaBalpha as well as an increase of nuclear NF-kappaB in both cells. ANE treatment did not activate epidermal growth factor receptor signaling system, but blockage of NF-kappaB activation rendered the suppression of ANE-modulated COX-2 upregulation in OECM-1. This study identified that ANE affected interactive signaling systems in oral keratonocytes that could be the pathogenetic basis for areca. PMID- 15825185 TI - Penile cancer: importance of circumcision, human papillomavirus and smoking in in situ and invasive disease. AB - Few population-based case-control studies have assessed etiologic factors for penile cancer. Past infection with high-risk human papillomavirus (HPV) is a known risk factor for penile cancer; however, few previous studies have related the HPV DNA status of the tumor to potential demographic and behavioral risk factors for the disease or evaluated whether in situ and invasive penile cancer share risk factors. Little information is available on the role and timing of circumcision in the etiology of penile cancer. We conducted a population-based case-control study in western Washington state that included 137 men diagnosed with in situ (n = 75) or invasive (n = 62) penile cancer between January 1, 1979, and December 31, 1998, and 671 control men identified through random digit dialing. Cases and controls were interviewed in person and provided peripheral blood samples. Case and control blood samples were tested for antibodies to HPV16 and HSV-2, and tumor specimens from cases were tested for HPV DNA. Men not circumcised during childhood were at increased risk of invasive (OR = 2.3, 95% CI 1.3-4.1) but not in situ (OR = 1.1, 95% CI 0.6-1.8) penile cancer. Approximately 35% of men with penile cancer who had not been circumcised in childhood reported a history of phimosis compared to 7.6% of controls (OR = 7.4, 95% CI 3.7-15.0). Penile conditions such as tear, rash and injury were associated with increased risk of disease. Among men not circumcised in childhood, phimosis was strongly associated with development of invasive penile cancer (OR = 11.4, 95% CI 5.0 25.9). When we restricted our analysis to men who did not have phimosis, the risk of invasive penile cancer associated with not having been circumcised in childhood was not elevated (OR = 0.5, 95% CI 0.1-2.5). Cigarette smoking was associated with a 4.5-fold risk (95% CI 2.0-10.1) of invasive penile cancer. HPV DNA was detected in 79.8% of tumor specimens, and 69.1% of tumors were HPV16 positive. The proportion of HPV DNA-positive tumors did not vary by any risk factors evaluated. Many risk factors were common for both in situ and invasive disease. However, 3 factors that did not increase the risk for in situ cancer proved significant risk factors for invasive penile cancer: lack of circumcision during childhood, phimosis and cigarette smoking. The high percentage of HPV DNA positive tumors in our study is consistent with a strong association between HPV infection and the development of penile cancer regardless of circumcision status. Circumcision in early childhood may help prevent penile cancer by eliminating phimosis, a significant risk factor for the disease. PMID- 15825186 TI - Neuromuscular manifestations of critical illness. AB - Critical illness, more precisely defined as the systemic inflammatory response syndrome (SIRS), occurs in 20%-50% of patients who have been on mechanical ventilation for more than 1 week in an intensive care unit. Critical illness polyneuropathy (CIP) and myopathy (CIM), singly or in combination, occur commonly in these patients and present as limb weakness and difficulty in weaning from the ventilator. Critical illness myopathy can be subdivided into thick-filament (myosin) loss, cachectic myopathy, acute rhabdomyolysis, and acute necrotizing myopathy of intensive care. SIRS is the predominant underlying factor in CIP and is likely a factor in CIM even though the effects of neuromuscular blocking agents and steroids predominate in CIM. Identification and characterization of the polyneuropathy and myopathy depend upon neurological examination, electrophysiological studies, measurement of serum creatine kinase, and, if features suggest a myopathy, muscle biopsy. The information is valuable in deciding treatment and prognosis. PMID- 15825187 TI - The relationship between adsorption energies of methyl on metals and the metallic electronic properties: a first-principles DFT study. AB - A theoretical study of CH3 adsorbed on the (111) surface of some transition and noble metal surfaces M (M = Cu, Ni, Rh, Pt, Pd, Ag, Au) and on the Fe(100) is presented. We find that the hollow site is preferred more than the top one for Fe, Ni, Rh, and Cu, but it is the other way for Pt, Pd, Au, and Ag. In addition, a good linear relationship was observed between the chemisorption energy and d band center for Group VIII metals or the square of the coupling matrix element for Group IB metals at the hollow site. Interestingly, with a detailed comparison of the adsorption energies at the top and hollow sites, we find that the adsorption energies among each group are very similar on the top site, which supports the theoretical model of Hammer and Norskov that the coupling between the HOMO of adsorbate and sp states of the metal is dominant and almost equal, and that the second coupling to the d-band contributes less but reflects the change of the adsorption energy. It confirms that the coupling to the d band comprises two opposite factors, that is, the d-band center was attractive and the square of the coupling matrix element was repulsive, such that the contributions from the two factors can counteract each other at the top site. PMID- 15825188 TI - Quantum chemical modeling of the reduction of cis-diammineplatinum(IV) tetrachloride [Pt(NH3)2Cl4] by methyl thiolate anion. AB - We present here both an ab initio and quantum mechanical/molecular mechanical (QM/MM) study of the cis-[Pt(NH3)2Cl4] complex reduction by methyl thiolate anion, SCH(-3), which is used as a model of glutathione. Geometry and electronic structure of cis-[Pt(NH3)2Cl4] are determined without and in aqueous medium. The mechanism of the reaction of reduction is characterized. The calculated activation energy of the reaction compares remarkably well with the experimental value. PMID- 15825189 TI - Role of Ser50 phosphorylation in SCG10 regulation of microtubule depolymerization. AB - Members of the stathmin-like protein family depolymerize microtubules (MTs), probably due to the ability of each stathmin monomer to bind two tubulin heterodimers in a complex (T(2)S complex). SCG10, a member of this family, is localized in the growth cone of neurons. It has four identified sites of serine phosphorylation (S50, S63, S73, and S97). Of these, S50 and S97 are phosphorylated by cAMP-dependent protein kinase, an enzyme involved in growth cone guidance. When the equivalent sites in stathmins are phosphorylated, they lose their ability to depolymerize MTs. We investigated the specific role of the two cAMP-dependent protein kinase (PKA) phosphorylation sites in SCG10. A mutant of SCG10 phosphorylated only on S50 retained the ability to depolymerize MTs, but SCG10 phosphorylated on S97 or on both S50 and S97 lost MT-depolymerizing activity. Surface plasmon resonance studies revealed that the phosphorylation of SCG10 at these sites reduced the tubulin heterodimer binding, mainly due to a reduced rate of association. In particular, compared to the two other phosphorylated forms, SCG10 phosphorylated at S50 had a significantly smaller dissociation constant for the binding of the first tubulin heterodimer and larger association and dissociation rate constants for the binding of the second heterodimer. This indicates that the phosphorylation of S50 compensates for the effect of phosphorylation at other sites by modulating T2S complex formation. Furthermore, these results suggest that S50-P maintains MT-depolymerizing activity, which indicates that the biological functions of phosphorylation at S50 and S97 are different. PMID- 15825190 TI - Early apoptotic and late necrotic components associated with altered Ca2+ homeostasis in a peptide-delivery model of polyglutamine-induced neuronal death. AB - The mechanisms by which polyglutamine expansion causes common features of neuronal death remain unclear. Here we describe an approach for delivering polyglutamine expansions directly into cultured sympathetic neurons. Glutamine (Q) residues (n = 10, 22, 30) were conjugated with a peptide possessing translocation properties across plasma membranes (PDP) and a nuclear localization signal (NLS). These peptides were rapidly incorporated into sympathetic neurons and showed neurotoxicity in a length- and dose-dependent manner. A robust induction of c-jun and cyclin D1 occurred following treatment with PDP-Q22-NLS. Enhanced c-Jun phosphorylation showed c-Jun N-terminal kinase (JNK) activation. Coincidentally, TrkA tyrosine phosphorylation was decreased in association with loss of phospho-Akt, the downstream target of PI-3 kinase. Despite such proapoptotic signals, neither release of cytochrome c from mitochondria nor caspase-3/7 activation was detected. TdT-mediated dUTP nick-end labeling-positive nuclear condensation, but no fragmentation, occurred. At 24 hr of treatment, cytoplasmic Ca2+ levels began to become elevated, and the cellular level of ATP was decreased. Cytoplasmic Ca2+ responses to KCl depolarization displayed a delayed recovery, providing evidence for lack of Ca2+ homeostasis. The neurons became committed to death at about 36 hr when mitochondrial Ca2+ uptake declined concurrently with loss of mitochondrial membrane potential. Collectively, these results show that, despite induction of early apoptotic signals, nonapoptotic neuronal cell death occurred via perturbed Ca2+ homeostasis and suggest that mitochondrial permeability transition may play important roles in this model of neuronal death. PMID- 15825191 TI - D-beta-hydroxybutyrate is neuroprotective against hypoxia in serum-free hippocampal primary cultures. AB - Hypoxia decreased survival of cultured rat primary hippocampal neurons in a time dependent manner. Addition of 4 mM Na D-beta-hydroxybutyrate (bHB), a ketone body, protected the cells for 2 hr and maintained the increase in survival compared to that of controls for up to 6 hr. Trypan blue exclusion indicated that acute cell death was reduced markedly after 2-hr exposure to hypoxia in the bHB treated group. The presence of bHB also decreased the number of neurons exhibiting condensed nuclei visualized by propidium iodide, indicative of apoptosis. The mitochondrial transmembrane potential (Em/c) was maintained for up to 2 hr exposure to hypoxia in the bHB-treated group, whereas the potential in the control group was decreased. Furthermore, cytochrome C release, caspase-3 activation, and poly (ADP-ribose) polymerase (PARP) cleavage were decreased in the bHB-treated group for the first 2 hr of exposure. These findings indicate that ketone bodies may be a candidate for widening the therapeutic window before thrombolytic therapy and at the same time decreasing apoptotic damage in the ischemic penumbra. PMID- 15825192 TI - Coregistration of quantitative proton magnetic resonance spectroscopic imaging with neuropathological and neurophysiological analyses defines the extent of neuronal impairments in murine human immunodeficiency virus type-1 encephalitis. AB - Relatively few immune-activated and virus-infected mononuclear phagocytes (MP; perivascular macrophages and microglia) may affect widespread neuronal dysfunction during human immunodeficiency virus type 1 (HIV-1)-associated dementia (HAD). Indeed, histopathological evidence of neuronal dropout often belies the extent of cognitive impairment. To define relationships between neuronal function and histopathology, proton magnetic resonance spectroscopic imaging (1H MRSI) and hippocampal long-term potentiation (LTP) were compared with neuronal and glial immunohistology in a murine model of HIV-1 encephalitis (HIVE). HIV-1(ADA)-infected human monocyte-derived macrophages (MDM) were stereotactically injected into the subcortex of severe combined immunodeficient (SCID) mice. Sham-operated and unmanipulated mice served as controls. Seven days after cell injection, brain histological analyses revealed a focal giant cell encephalitis, with reactive astrocytes, microgliosis, and neuronal dropout. Strikingly, significant reductions in N-acetyl aspartate concentration ([NAA]) and LTP levels in HIVE mice were in both injected and contralateral hemispheres and in brain subregions, including the hippocampus, where neuropathology was limited or absent. The data support the importance of 1H MRSI as a tool for assessing neuronal function for HAD. The data also demonstrate that a highly focal encephalitis can produce global deficits for neuronal function and metabolism. PMID- 15825193 TI - Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells. AB - Embryonic stem (ES) cells proliferate and maintain their pluripotency for over 1 year in vitro and may therefore provide a sufficient source for cell therapies. However, most of the previously reported methods for obtaining a source for cell therapies have not been simple. We describe here a novel method for induction of neurospheres from mouse ES cells by coculturing on PA6 cells instead of the formation of embryoid bodies. The ES cells cocultured with the PA6 stromal cell line for at least 3 days were capable of differentiating into spheres. The cells in the spheres were all green fluorescent protein (GFP) positive, showing that they were derived from GFP-expressing D3-ES cells. The spheres contained nestin positive cells. The number of spheres increased when they were cocultured with PA6 for a longer period. Sphere formation was observed even after 10 mechanical dissociations and subculturings, showing its self-renewal ability. The cells differentiated into microtubule-associated protein-2 (MAP2)-positive neuronal cells and glial fibrillary acidic protein (GFAP)-positive glial cells. gamma Aminobutyric acid-positive cells and tyrosine hydroxylase-positive cells were also observed in the spheres. The percentages of the MAP2- or GFAP-positive cells in the sphere changed according to the period of coculture on PA6 cells. At an early stage of coculture, more neurons were generated and, at a later period, more glial cells were generated. These results suggested that neurosphere could be generated from ES cells by coculturing with PA6, and that these cells resembled neural stem cells derived from mouse fetal brain tissue. PMID- 15825194 TI - Caspases-2, -3, and -7 are involved in thapsigargin-induced apoptosis of SH-SY5Y neuroblastoma cells. AB - Caspase-2 has been reported to play a role in the cell death observed under a number of different conditions; however, it is unclear whether caspase-2 plays a role in cell death triggered by endoplasmic reticulum (ER) stress. The purpose of this study was to determine whether caspase-2 is involved in SH-SY5Y neuroblastoma cell death caused by thapsigargin-induced ER stress. Thapsigargin treatment (1 microM, 16 hr) stimulated the proteolytic processing of caspases-2, 3, and -7, suggesting that these caspases are activated by ER stress. The role of these caspases in thapsigargin-induced cell death was examined by using cell permeable caspase inhibitors. In the absence of pretreatment with caspase inhibitors, thapsigargin (0.1 microM, 20 hr) reduced the number of viable cells to 53.9% +/- 3.3% of starting-time control. Pretreatment for 90 min with either the pan-caspase inhibitor Z-VAD-FMK or the caspase-2-selective inhibitor Z-VDVAD FMK inhibited thapsigargin-stimulated cell death, resulting in the number of viable cells being 115.6% +/- 5.3% (P < 0.001) and 69.3% +/- 2.9% (P < 0.01), respectively, of starting-time control. Neither the caspase-3- and -7-selective inhibitor Z-DEVD-FMK nor the caspase-9-selective inhibitor Z-LEHD-FMK significantly affected thapsigargin-stimulated cell death. An anticaspase-12 reactive protein was also identified in SH-SY5Y cells, but thapsigargin had no effect on proteolysis of this protein. These data demonstrate that caspases-2, 3, and -7 are involved in ER stress-mediated death of SH-SY5Y cells. PMID- 15825196 TI - Stereoselective kinetic study of hexaconazole enantiomers in the rabbit. AB - Hexaconazole [(RS)-2-(2,4-dichlorophenyl)-1-(1H-1,2,4-triazol-1-yl)hexan-2-ol] is a potent triazole fungicide. The (-) isomer accounts for most of the fungicidal activity. The stereo- and/or enantioselective kinetics of hexaconazole were investigated in rabbits by intravenous injection. The concentrations of (-)- and (+)-hexaconazole in plasma, liver, and kidney tissue were determined by HPLC with a cellulose tris(3,5-dimethylphenylcarbamate)-based chiral stationary phase and by gas chromatography-mass spectrometry. After intravenous administration of racemic hexaconazole (rac-hexaconazole) at 30 mg/kg, plasma, liver, and kidney levels of the (+)-enantiomer decreased more rapidly than those of the (-) enantiomer. The (-)-/(+)-enantiomer ratio of the area under the concentration time curve (AUC(0-infinity)) was 1.35. The total plasma clearance value (CL) of (+)-enantiomer was more than 1.3-fold higher than that of the (-)-hexaconazole. The enantiomeric ratio (ER) increased with time in plasma, liver, and kidney. Other pharmacokinetic parameters of the enantiomers were also different. These results indicate substantial stereoselectivity in the kinetics of hexaconazole enantiomers in rabbits. PMID- 15825197 TI - Configurational stability of 2-benzoylcyclohexanone: unexpected solvent effects on the rate of racemization. AB - The kinetics of the racemization of 2-benzoylcyclohexanone 1 in hexanes, ethanol, and mixtures thereof have been investigated by time dependence of enantiomeric purity using enantioselective HPLC. In pure hexanes and ethanol, the racemization half-lives were determined as 552 and 23.8 min, respectively, at 66 degrees C. Surprisingly, racemization of 1 in mixtures of hexanes and ethanol was found to involve an induction period followed by a sigmoidal decrease of the enantiomeric excess with half-lives varying between 11.5 and 24.0 min. This unexpected solvent influence on the rate of racemization of 1 was attributed to complex isomerization mechanisms involving three possibly interconverting enol tautomers of 1. PMID- 15825198 TI - Identifying stereoisomers of the asymmetric hydrogenation catalyst [Me-BPE Rh(COD)]+BF4 -. AB - The performance of a catalyst used in asymmetric synthesis is likely to be dependent upon its stereoisomeric purity. An impurity was detectable by (31)P NMR in early development batches of the asymmetric hydrogenation catalyst [(S,S)-Me BPE-Rh(COD)](+)BF(4) (-) made from the ligand bis((2S,5S)-2,5 dimethylphospholano)ethane [(S,S)-Me-BPE]. Its identity as a stereoisomer with one chiral and one meso-phospholane ring was deduced by comparison of the (31)P NMR spectra and GC traces of the ligand with a deliberately synthesized mixture of isomers. Interestingly, the impurity corresponded to a trans-meso isomer formed by thermal (200 degrees C) pyramidal inversion at phosphorus of the initially synthesized cis-meso-phospholane when the ligand was purified by distillation. Low levels of this trans-meso/chiral impurity do not significantly impair the enantioselectivity of the rhodium complex as an asymmetric hydrogenation catalyst, but high levels of stereochemical impurities resulted in a loss of both enantioselectivity and activity. Therefore it is indeed important to establish that a catalyst used in asymmetric catalysis is sufficiently stereoisomerically pure. Owing to strict control of the stereochemical purity of the key hexane-2,5-diol intermediate, the impurity is not detected in production batches. PMID- 15825199 TI - Effect of dairy products on the lifetime of Provox2 voice prostheses in vitro and in vivo. AB - BACKGROUND: Reduction of biofilm formation on tracheoesophageal voice prostheses by certain dairy products might extend their clinical lifetime. The purpose of this study was to determine the influence of certain dairy products on voice prosthetic biofilms and lifetimes in vitro and in vivo. METHODS: The in vitro results were accomplished using an artificial throat. The lifetimes of Provox2 prostheses were evaluated in a patient group that daily consumed the evaluated products. RESULTS: Buttermilk and Yakult Light fermented milk decreased the amount of bacteria on voice prostheses but stimulated yeast prevalence in vitro. Concurrently, lifetimes of voice prostheses in patients consuming buttermilk were not significantly different, whereas patients consuming Yakult Light fermented milk drink had a significantly (p < .01) increased prosthesis lifetime by a factor of 3.76. CONCLUSION: Yakult Light fermented milk drink reduced biofilm formation on Provox2 prostheses in vitro and in vivo and significantly increased prosthesis lifetime. In vivo, no significant effects were observed for patients consuming buttermilk. PMID- 15825200 TI - A case of cervical metastases from temporal bone carcinoid. AB - BACKGROUND: Carcinoids are neuroendocrine tumors that typically occur in the gastrointestinal tract and lung and less frequently in the head and neck region. Whereas the metastatic potential of laryngeal carcinoid is well documented, only one case of metastasis of temporal bone carcinoid to the cervical lymph nodes has been reported. We present the case of a 55-year-old woman with cervical metastases almost 3 years after a primary diagnosis of temporal bone carcinoid. METHODS: The patient had undergone a subtotal petrosectomy for a temporal bone carcinoid and, 29 months later, revision surgery for a local recurrence. Four months later, multiple laterocervical swelling appeared, and the patient underwent modified radical neck dissection. RESULTS: Histologic examination after the neck dissection was suggestive of lymph node metastases from a carcinoid tumor. Immunohistochemical analysis of the lymph node metastases showed neoplastic cells positive for keratin, vimentin, chromogranin, and neuron specific enolase and negative for thyroglobulin, calcitonin, S-100, and parathyroid hormone. The patient was treated with intensity-modulated radiation therapy and neck irradiation. CONCLUSIONS: We believe that temporal bone carcinoids have metastatic potential not predictable by histologic features. Surgical excision is the treatment of choice for patients with temporal bone carcinoids, with the approach and technique determined by the extent of the mass. Moreover, before planning surgery and during follow-up, neck node status must be carefully detected. PMID- 15825202 TI - Head and neck cancer care at a community-based teaching hospital. AB - BACKGROUND: The purpose of this analysis was to demonstrate that the National Cancer Institute (NCI) Community Clinical Oncology Program (CCOP) will result in accruals to national research protocols for patients with malignancies in a community-based teaching hospital. METHODS: We retrospectively reviewed the records of oncology patients eligible for clinical trials referred to the Helen F. Graham Cancer Center (HFGCC). RESULTS: The average number of patients who actually enroll in clinical trials nationally is 2.5%. The number of patients who enrolled in clinical trials at the HFGCC CCOP in 2003 was 24%. CONCLUSIONS: The integration of a CCOP in a community-based teaching hospital will enhance adoption of state-of-the-art care and increase accruals to national research protocols. PMID- 15825201 TI - Craniofacial resection for malignant paranasal sinus tumors: Report of an International Collaborative Study. AB - BACKGROUND: Malignant tumors of the superior sinonasal vault are rare, and, because of this and the varied histologic findings, most outcomes data reflect the experience of small patient cohorts. This International Collaborative study examines a large cohort of patients accumulated from multiple institutions experienced in craniofacial surgery, with the aim of reporting benchmark figures for outcomes and identifying patient-related and tumor-related predictors of prognosis after craniofacial resection (CFR). METHODS: Three hundred thirty-four patients from 17 institutions were analyzed for outcome. Patients with esthesioneuroblastoma were excluded and are being reported separately. The median age was 57 years (range, 3-98 years). One hundred eighty-eight patients (56.3%) had had prior single-modality or combined treatment, which included surgery in 120 (36%), radiation in 79 (23.7%), and chemotherapy in 56 (16.8%). The most common histologic findings were adenocarcinoma in 107 (32%) and squamous cell carcinoma in 101 (30.2%). The margins of resection were close or microscopically positive in 95 (30%). Adjuvant radiotherapy was given in 161 (48.2%) and chemotherapy in 16 (4.8%). Statistical analyses for outcomes were performed in relation to patient characteristics, tumor characteristics, including histologic findings and extent of disease, surgical resection margins, prior radiation, and prior chemotherapy to determine predictive factors. RESULTS: Postoperative mortality occurred in 15 patients (4.5%). Postoperative complications occurred in 110 patients (32.9%). The 5-year overall, disease-specific, and recurrence-free survival rates were 48.3%, 53.3%, and 45.8%, respectively. The status of surgical margins, histologic findings of the primary tumor, and intracranial extent were independent predictors of overall, disease-specific, and recurrence-free survival on multivariate analysis. CONCLUSIONS: CFR for malignant paranasal sinus tumors is a safe surgical treatment with an overall mortality of 4.5% and complication rate of 33%. The status of surgical margins, histologic findings of the primary tumor, and intracranial extent are independent predictors of outcome. PMID- 15825203 TI - AMORE protocol in pediatric head and neck rhabdomyosarcoma: descriptive analysis of failure patterns. AB - BACKGROUND: The AMORE protocol is a local treatment for patients with nonorbital pediatric head and neck rhabdomyosarcoma (HNRMS). The objectives of this study were: (1) to assess the adequacy of the concept, and (2) to identify factors associated with relapse. METHODS: We performed a retrospective multidisciplinary review of 22 children primarily treated according to the AMORE protocol, excluding two children with inadequate imaging data. RESULTS: Seven patients had a local relapse, six within and one outside the residual tumor area. Five of the six patients with relapse in the residual area had gross total or debulking (incomplete) surgery, suboptimal position of the mold for brachytherapy, or both. In the 15 nonrecurrent cases, four patients had either incomplete surgery or suboptimal mold position. Both surgical and brachytherapeutic factors seem to be associated with relapse. CONCLUSIONS: AMORE is an adequate concept. More rigid preoperative imaging and intraoperative verification of the brachytherapy mold position might lead to a reduction in the number of local failures. PMID- 15825204 TI - Advanced squamous cell carcinoma of lower gingivobuccal complex: patterns of spread and failure. AB - BACKGROUND: Carcinoma of the gingivobuccal complex is commonly associated with the use of smokeless tobacco known as "quid." METHODS: We conducted a retrospective chart review of 511 patients with advanced cancer of gingivobuccal complex surgically treated during 1994 to 1995. We evaluated patterns of disease failure in these patients and correlated disease-free survival with various prognostic factors. RESULTS: During a median follow-up of 46 months, 159 locoregional recurrences and 11 distant metastases were detected in 148 patients. Seventy-nine percent of the recurrences appeared within 18 months of surgery, and the median survival for patients with recurrent disease was less than 4 months. Two-year and 5-year disease-free survival rates were 64% and 57%, respectively. On multivariate analysis, disease-free survival showed significant correlation with skin involvement and extracapsular spread. CONCLUSIONS: Gingivobuccal cancers usually fail locoregionally. Soft tissue infiltration and extracapsular spread of nodal disease influence disease-free survival. PMID- 15825205 TI - Complications of craniofacial resection for malignant tumors of the skull base: report of an International Collaborative Study. AB - BACKGROUND: Advances in imaging, surgical technique, and perioperative care have made craniofacial resection (CFR) an effective and safe option for treating malignant tumors involving the skull base. The procedure does, however, have complications. Because of the relative rarity of these tumors, most existing data on postoperative complications come from individual reports of relatively small series of patients. This international collaborative report examines a large cohort of patients accumulated from multiple institutions with the aim of identifying patient-related and tumor-related predictors of postoperative morbidity and mortality and set a benchmark for future studies. METHODS: One thousand one hundred ninety-three patients from 17 institutions were analyzed for postoperative mortality and complications. Postoperative complications were classified into systemic, wound, central nervous system (CNS), and orbit. Statistical analyses were carried out in relation to patient characteristics, extent of disease, prior radiation treatment, and type of reconstruction to determine factors that predicted mortality or complications. RESULTS: Postoperative mortality occurred in 56 patients (4.7%). The presence of medical comorbidity was the only independent predictor of mortality. Postoperative complications occurred in 433 patients (36.3%). Wound complications occurred in 237 (19.8%), CNS-related complications in 193 (16.2%), orbital complications in 20 (1.7%), and systemic complications in 57 (4.8%) patients. Medical comorbidity, prior radiation therapy, and the extent of intracranial tumour involvement were independent predictors of postoperative complications. CONCLUSIONS: CFR is a safe surgical treatment for malignant tumors of the skull base, with an overall mortality of 4.7% and complication rate of 36.3%. The impact of medical comorbidity and intracranial tumor extent should be carefully considered when planning therapy for patients whose tumors are amenable to CFR. PMID- 15825206 TI - Measurement of the coagulation dynamics of bovine liver using the modified microscopic Beer-Lambert law. AB - BACKGROUND AND OBJECTIVES: During heating, the optical properties of biological tissues change with the coagulation state. In this study, we propose a technique, which uses these changes to monitor the coagulation process during laser-induced interstitial thermotherapy (LITT). STUDY DESIGN/MATERIALS AND METHODS: Untreated and coagulated (water bath, temperatures between 35 degrees C and 90 degrees C for 20 minutes.) samples of bovine liver tissue were examined using a Nd:YAG (lambda = 1064 nm) frequency-domain reflectance spectrometer. We determined the time integrated intensities (I(DC)) and the phase shifts (Phi) of the photon density waves after migration through the tissue. From these measured quantities, the time of flight (TOF) of the photons and the absorption coefficients of the samples were derived using the modified microscopic Beer-Lambert law. RESULTS: The absorption coefficients of the liver samples decreased significantly with the temperature in the range between 50 degrees C and 70 degrees C. At the same time, the TOF of the investigated photos was found increased indicating an increased scattering. The coagulation dynamics could be well described using the Arrhenius formalism with the activation energy of 106 kJ/mol and the frequency factor of 1.59 x 10(13)/second. CONCLUSIONS: Frequency-domain reflectance spectroscopy in combination with the modified microscopic Beer-Lambert (MBL) is suitable to measure heat induced changes in the absorption and scattering properties of bovine liver in vitro. The technique may be used to monitor the coagulation dynamics during local thermo-coagulation in vivo. PMID- 15825207 TI - Tensile bond strength of a flowable composite resin to ER:YAG-laser-treated dentin. AB - BACKGROUND AND OBJECTIVES: This in vitro study evaluated the influence of a flowable composite resin (FCR) on the tensile bond strength of resin to dentin treated with the Er:YAG Laser (L) and diamond bur (DB). STUDY DESIGN/MATERIALS AND METHODS: Ninety dentin surfaces obtained from 45 third molars were ground and randomly divided into six groups (n = 15): G1-DB, G2-DB+FCR, G3-L (100 mJ, 10 Hz, 37.04 J/cm2), G4-L (100 mJ, 10 Hz, 37.04 J/cm2)+FCR, G5-L (250 mJ, 2 Hz, 92.60 J/cm2), and G6-L (250 mJ, 2 Hz, 92.60 J/cm2)+FCR. After surface etching with 37% phosphoric acid and the application of an adhesive system, inverted conical specimens were prepared with a hybrid composite resin. In groups G2, G4, and G6 a FCR was placed before the hybrid composite resin. After 24 hours-storage in distilled water, the tensile test was performed in a universal testing machine (0.5 mm/minute, 500 N). RESULTS: Data were submitted to Kruskal Wallis test (P = 0.01). The mean bond strength values (MPa+/-SD) were: G1-13.54 (+/-2.99), G2 14.67 (+/-2.32), G3-9.49 (+/-3.09), G4-14.60 (+/-2.76), G5-8.97 (+/-3.89), and G6 13.02 (+/-2.18). Groups G1 and G2 presented the highest bond strength values, which were statistically similar to those of G4 and G6. The groups treated with laser and without the FCR (G3 and G5) showed the lowest shear bond strength values. CONCLUSIONS: FCR can increase the adhesion to dentin treated with Er:YAG laser within different parameters. PMID- 15825208 TI - Relevance vector machine for optical diagnosis of cancer. AB - BACKGROUND AND OBJECTIVES: A probability-based, robust diagnostic algorithm is an essential requirement for successful clinical use of optical spectroscopy for cancer diagnosis. This study reports the use of the theory of relevance vector machine (RVM), a recent Bayesian machine-learning framework of statistical pattern recognition, for development of a fully probabilistic algorithm for autofluorescence diagnosis of early stage cancer of human oral cavity. It also presents a comparative evaluation of the diagnostic efficacy of the RVM algorithm with that based on support vector machine (SVM) that has recently received considerable attention for this purpose. STUDY DESIGN/MATERIALS AND METHODS: The diagnostic algorithms were developed using in vivo autofluorescence spectral data acquired from human oral cavity with a N(2) laser-based portable fluorimeter. The spectral data of both patients as well as normal volunteers, enrolled at Out Patient department of the Govt. Cancer Hospital, Indore for screening of oral cavity, were used for this purpose. The patients selected had no prior confirmed malignancy and were diagnosed of squamous cell carcinoma (SCC), Grade-I on the basis of histopathology of biopsy taken from abnormal site subsequent to acquisition of spectra. Autofluorescence spectra were recorded from a total of 171 tissue sites from 16 patients and 154 healthy squamous tissue sites from 13 normal volunteers. Of 171 tissues sites from patients, 83 were SCC and the rest were contralateral uninvolved squamous tissue. Each site was treated separately and classified via the diagnostic algorithm developed. Instead of the spectral data from uninvolved sites of patients, the data from normal volunteers were used as the normal database for the development of diagnostic algorithms. RESULTS: The diagnostic algorithms based on RVM were found to provide classification performance comparable to the state-of-the-art SVMs, while at the same time explicitly predicting the probability of class membership. The sensitivity and specificity towards cancer were up to 88% and 95% for the training set data based on leave- one-out cross validation and up to 91% and 96% for the validation set data. When implemented on the spectral data of the uninvolved oral cavity sites from the patients, it yielded a specificity of up to 91%. CONCLUSIONS: The Bayesian framework of RVM formulation makes it possible to predict the posterior probability of class membership in discriminating early SCC from the normal squamous tissue sites of the oral cavity in contrast to dichotomous classification provided by the non-Bayesian SVM. Such classification is very helpful in handling asymmetric misclassification costs like assigning different weights for having a false negative result for identifying cancer compared to false positive. The results further demonstrate that for comparable diagnostic performances, the RVM-based algorithms use significantly fewer kernel functions and do not need to estimate any hoc parameters associated with the learning or the optimization technique to be used. This implies a considerable saving in memory and computation in a practical implementation. PMID- 15825209 TI - Preferences of Women Evaluating Risks of Tamoxifen (POWER) study of preferences for tamoxifen for breast cancer risk reduction. AB - BACKGROUND: The objective of this study was to understand the attitudes and preferences of risk-eligible women regarding use of tamoxifen for breast cancer risk reduction. METHODS: A cross-sectional, mixed-methods interview study was conducted at a university medical center and at community sites. Participants were women who had an estimated 5-year breast cancer risk > or = 1.7% and no prior breast cancer. Interviews were conducted in English or Spanish. The interview included a 15-minute, standardized educational session on the potential benefits and harms of tamoxifen followed by close-ended and open-ended questions about participants' inclinations to take tamoxifen and factors important to their decision. A demographic questionnaire, a test on knowledge of potential benefits and harms of tamoxifen, and an interview evaluation were included. RESULTS: Two hundred fifty-five women completed interviews. Their estimated mean 5-year breast cancer risk was 2.8%; and their mean self-perceived 5-year risk was 32.7%. After the educational intervention, 45 women (17.6%) were inclined to take tamoxifen. Very high risk women (> 3.5%) were no more inclined to take it than women with lower risk (1.7-3.5%). In a multivariable analysis, lower income, confidence in the effectiveness of tamoxifen, and concern about fractures were associated with being inclined to take it; concern about pulmonary embolism, dyspareunia, cataracts, and low self-perceived breast cancer risk were associated negatively with taking tamoxifen. Participants expressed concerns about adverse effects. CONCLUSIONS: Less than 20% of women were interested in tamoxifen after education about potential benefits and harms, despite a very high self-perceived breast cancer risk. Candidate chemoprevention agents must have few potential adverse effects to achieve widespread acceptance. PMID- 15825210 TI - Relation between smoking cessation and receiving results from three annual spiral chest computed tomography scans for lung carcinoma screening. AB - BACKGROUND: The relation between undergoing a single computed tomography (CT) screening for lung carcinoma and the potential long-term impact on smoking status has been equivocal. Perhaps, recommendations from multiple cancer screenings may promote smoking abstinence among individuals at high risk for lung carcinoma. METHODS: The current longitudinal study comprised 926 current smokers and 594 former smokers who participated in 3 annual follow-up low-dose, fast spiral chest CT scan screenings for lung carcinoma. Baseline demographic, pulmonary function, smoking history variables, and previous abnormal findings were evaluated as predictors of self-reported point prevalence smoking abstinence. RESULTS: Among current smokers at baseline, abstinence from smoking during the 3-year follow-up was associated with older age, worse baseline pulmonary function, and abnormal CT finding the previous year requiring interim follow-up. Of participants who received abnormal screens each of the previous 3 years, 41.9% reported smoking abstinence compared with 28.0% with 2 abnormal screens, 24.2% with 1 abnormal screen, and 19.8% with no abnormal screens. Among former smokers, abstinence from smoking was associated with a longer duration of abstinence before the baseline visit. CONCLUSIONS: Smokers with abnormal CT findings from multiple CT screens were more likely to be abstinent from smoking at the 3-year follow-up. Multiple low-dose, fast spiral chest CT scan screenings for lung carcinoma may represent teachable moments and opportunities to enhance motivation for smoking abstinence. Further research is needed to continue to investigate how annual screening may enhance motivation for health behavior change. PMID- 15825212 TI - Copper-catalyzed enantioselective conjugate addition of Grignard reagents to alpha,beta-unsaturated esters. PMID- 15825211 TI - Temporary dermal scatter reduction: quantitative assessment and implications for improved laser tattoo removal. AB - BACKGROUND AND OBJECTIVES: Temporary dermal clearing, i.e., reduction in the attenuation coefficient of the dermis and epidermis, may lead to improved laser tattoo removal by providing increased efficiency of laser delivery to embedded ink particles and enabling the use of shorter wavelength visible lasers more effective on certain inks. STUDY DESIGNS/MATERIALS AND METHODS: In a hairless guinea pig model of human tattoo, we tested both intradermal and transdermal application of glycerol, using visual inspection, spectral analysis, and optical coherence tomography techniques to assess effectiveness. In controlled experiments, we compared the outcomes of single laser treatment sessions for both cleared and uncleared tattoo sites using Q-switched 755 and 532 nm lasers on three different inks. RESULTS: Intradermal injection of clearing agents induced dermal clearing but resulted in necrosis and scar. Transdermal application of clearing agents resulted in moderate reversible clearing, which was localized to the superficial layers of the skin and did not result in complications. Statistically significant differences in laser treatment outcome were observed relative to a number of treatment parameters including the treatment of certain tattoos by short wavelength lasers. CONCLUSIONS: Temporary clearing of superficial skin layers may be performed in an apparently safe and reliable manner. Clearing should lead to increased penetration of laser light to tattoos and should, therefore, increase treatment efficiency. Further study is needed to determine the degree to which this change is of clinical value. PMID- 15825213 TI - Synthesis of diazirinyl photoprobe carrying a novel cleavable biotin. PMID- 15825214 TI - Quantum tunneling of magnetization in lanthanide single-molecule magnets: bis(phthalocyaninato)terbium and bis(phthalocyaninato)dysprosium anions. PMID- 15825215 TI - Vapochromic behavior accompanied by phase transition between charge-polarization and charge-density-wave states in a quasi-one-dimensional iodine-bridged dinuclear platinum compound. PMID- 15825216 TI - Separation of hyaluronic acid by ultrahigh-voltage capillary gel electrophoresis. AB - Hyaluronic acid was separated using 95 kV applied potential in a polyacrylamide gel-filled capillary. The results of this separation were compared to those obtained using a capillary electrophoresis instrument operated at a more conventional potential of 15 kV. For lower-molecular-weight oligomers, the separation efficiency was found to improve by about tenfold, and the resolution by about threefold. However, the improvement in resolution declined as the polymer molecular weight increased. PMID- 15825217 TI - Prediction of electrophoretic mobilities of peptides in capillary zone electrophoresis by quantitative structure-mobility relationships using the Offord model and artificial neural networks. AB - The aim of this work was to explore the usefulness of empirical models and multivariate analysis techniques in predicting electrophoretic mobilities of small peptides in capillary zone electrophoresis (CZE). The data set consists of electrophoretic mobilities, measured at pH 2.5, for 125 peptides ranging in size between 2 and 14 amino acids. Among the existing empirical models, the Offord model (i.e., mu identical with Q/M(2/3)) gave the best correlation for the data set. A quantitative structure-mobility relationship (QSMR) was developed using the Offord's charge-over-mass term (Q/M(2/3)) as one descriptor combined with the corrected steric substituent constant (E(s, c)) and molar refractivity (MR) descriptors to account for the steric effects and bulkiness of the amino acid side chains. The multilinear regression (MLR) of the data set showed an improvement in the predictive ability of the model over the simple Offord's relationship. A 3-4-1 back propagation artificial neural networks (BP-ANN) model resulted in a significant improvement in the predictive ability of the QSMR over the MLR treatment, especially for peptides of higher charges that contain basic amino acids arginine, histidine, and lysine. The improved correlations by the BP ANN analysis suggest the existence of nonlinear characteristic in the mobility charge relationships. PMID- 15825218 TI - Electroosmotic flow suppression in capillary electrophoresis: chemisorption of trimethoxy silane-modified polydimethylacrylamide. AB - Adsorbed polymers are widely used to suppress electroosmotic flow (EOF) in capillary electrophoresis (CE). Polymeric coatings, physisorbed onto the surface of the capillary wall, are often unstable under harsh conditions. This can be attributed to the reversible nature of the coating which becomes apparent when the adsorbed layer competes with a second species in the electrophoresis buffer solution for attachment/interaction with the capillary surface. In an effort to overcome the problem of coating instability, trimethoxysilane-modified polydimethylacrylamide was synthesized. This copolymer rapidly adsorbs on the wall from ultradilute aqueous solutions. After incubation at a temperature of 60 degrees C silyl groups, which extend from the polymer backbone, form condensation bonds with the silanols on the capillary surface. This enables subsequent formation of strong covalent bonds between the copolymer and the capillary wall. In this research, we establish that physisorption of polymer chains to the surface is essential for close alignment of surface and polymer silane groups which facilitates the formation of covalent bonds. PMID- 15825219 TI - Relationship between biochemical biomarkers and pre-copulatory behaviour and mortality in Gammarus pulex following pulse-exposure to lambda-cyhalothrin. AB - The impact of the pyrethroid lambda-cyhalothrin was studied in an in-stream mesocosm placed in a natural riffle of a Danish stream. Twice during summer 2002, the natural macroinvertebrate community was exposed in situ to a 30-min pulse of lambda-cyhalothrin. During exposure, nets caught macroinvertebrates in drift. Exposed and unexposed individuals of the amphipod Gammarus pulex (L) were transferred to aquaria in the laboratory and monitored for biochemical changes (ie biomarkers), pre-copulatory behaviour and mortality. Biochemical biomarkers were identified by screening extracts of exposed and unexposed G pulex using high performance planar chromatography (HPPC). Biochemical biomarkers were detected 3 h after pulse-exposure, and one biomarker was persistent up to 7 days after exposure. Pre-copulatory behaviour (ie pair formation) was significantly impaired up to 5 days after exposure, and had not fully recovered at the end of the observation period. EC10(0.5 h) and EC50(0.5 h) values for pre-copulatory behaviour were 0.04 and 0.20 microg litre(-1), respectively. Mortality was significant at 0.35 microg litre(-1) with an LC50(0.5 h) of 5.69 microg litre( 1). There was a significant relationship between two biomarkers and mortality. The study demonstrated that pulse-exposure at expected environmental concentrations can reduce local populations of G pulex, resulting in severe impact on populations with limited possibilities of re-colonisation. PMID- 15825220 TI - Piperazine-based buffers for liposome coating of capillaries for electrophoresis. AB - Anionic liposomes can be coated on fused-silica capillaries for electrophoresis in the presence of N-(hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES) as background electrolyte (BGE) solution. In this work, the interaction of various compounds with zwitterionic and anionic phospholipid coatings was studied with HEPES at pH 7.4 as BGE solution. The chromatographic and electrophoretic behavior of three test sample solutions (anionic, cationic, and neutral) was investigated for evaluation of the phospholipid coatings. Our results show that hydrophobic interactions between analytes and the phospholipid coating are important for the migration of charged analytes. In addition, the performances of other piperazine-based buffers, i.e., N-(2-hydroxyethyl)piperazine-N'-(2 hydroxypropanesulfonic acid), piperazine-N,N'-bis(2-ethanesulfonic acid), and piperazine-N,N'-bis(hydroxypropane sulfonic acid), at pH 7.4, as liposome solvent and BGE solution were evaluated and compared with the performance of HEPES at pH 7.4. The anionic liposome solution comprised 80/20 mol% phosphatidylcholine/phosphatidylserine. A simple test solution was selected and the chromatographic and electrophoretic migration behavior of the analytes was evaluated. The results show that, in addition to HEPES, other piperazine-based buffers at pH 7.4 are suitable for coating of fused-silica capillaries with anionic liposomes. PMID- 15825221 TI - Bibliography. Current world literature. Respiratory infection. PMID- 15825222 TI - Cell biology. GSK-3beta and microtubule assembly in axons. PMID- 15825223 TI - Patient information series. Chronic obstructive pulmonary disease (COPD). PMID- 15825224 TI - [A course of idiopathic generalized epilepsy in adult and elderly patients]. AB - Thirty-six patients with idiopathic generalized epilepsy (IGE) aged 35-67 years (mean age 44.4 +/- 10.6 years, 14 male, 22 female) have been studied. Ten of them were over 50 years, 8 had a diagnosis of juvenile absence epilepsy (JAE); 12- juvenile myoclonic epilepsy (JME) and 16--awakening epilepsy (AE), IGE with isolated generalized tonic-clonic seizures. The control group included 80 patients with IGE (22 male, 58 female aged 15-34 years, mean age 20.5 +/- 4.3 years), 29 of them being affected with JAE, 28--JME, 23--AE. Elderly patients with IGE experienced difficulties in the control over absences, myoclonic seizures and generalized seizures. In this group, the seizures-free cases were rare. The results suggest worse clinical prognosis of IGE for elderly patients as compared with young ones. PMID- 15825225 TI - [Clinical aspects of witchcraft delusions]. AB - To distinguish clinical variants and to specify nosologic entity of witchcraft delusions, 69 patients (10 males, aged 15-72 years) have been examined. It was found that witchcraft delusions exist in passive and active forms. In a passive form, the patient is sure that unknown (mystic) power damaged him/her; in an active form the patient, possessing a gift for unusual abilities, can influence the others (bewitches, heals, etc). Five clinical syndromes, in the structure of which the above delusions were found, namely, paranoiac-hypochondriac, hallucination-paranoid, depressive-paranoid, paraphrenic and delirious, were identified. Psychoses of schizophrenia spectrum were diagnosed in 52 patients, organic--in 8, alcoholic--in 7 and recurrent depressive disorder--in 2. Clinical significance of witchcraft delusions is closely related to its social aspect. Being combined with ideas of persecution, poisoning and damage, it results in the brutal forms of delusions defense and may be considered as an unfavorable prognostic trait. PMID- 15825226 TI - [Parameters of platelet serotonin system in patients with schizophrenia treated with olanzapine: a search for the serotonergic predictors of therapeutic efficacy]. AB - Fifty-nine patients, 21 women and 38 men, with ICD-10 diagnosis of schizophrenia (F20.0), attack-like type, were treated with olanzapine during 28 weeks (8-weeks of acute and 20-weeks of maintenance therapy). Evaluation of clinical symptoms measured by the Positive and Negative Syndromes scale (PANSS) revealed that female patients responded better to therapy as compared to male ones, with PANSS total, PANSS negative and PANSS general psychopathological scores being significantly reduced (p < 0.006) in females after 1 week of the treatment and in males--after 2 weeks. In the female group, a reduction of PANSS total score by 50% in the acute stage of treatment qualified as a very good response was observed in 7 (33%) patients and in the male group--in 1 (2.7%). The between groups difference was highly significant (p = 0.002). When examined for a rate of 3H-serotonin uptake into platelets, density of sites of 3H-imipramine binding on the whole platelets, platelet serotonin level and levels of high- also low molecular weight forms of platelet immunoreactive serotonin transporter protein, a significant decrease of the platelet serotonin level, comparing to controls, was detected in the female group before treatment. During the treatment, this parameter gradually increased up to control level. Other parameters did not change significantly for 28-weeks of therapy and did not differ from the control values. There were positive correlations between the levels of platelet serotonin before treatment and subsequent reduction of the PANSS total and positive subscale scores in the female group. In responders with a very good treatment related response, the serotonin level in the platelets before treatment was higher compared to the values in resistant patients: 5.4 +/- 2.5 and 2.7 +/- 1.3 nmol/10(9) cells, respectively. Relative risk (RR) of unfavorable treatment outcome in patients with initially reduced levels of platelet serotonin was approximately twice lower (RR = 1.83; Cl 95% 1.1-34.9) than that in patients with normal or elevated levels of platelet serotonin. The results suggest that selection of patients with initial higher level of platelet serotonin before olanzapine treatment can reduce the risk of non-responding to therapy by 36%. PMID- 15825227 TI - [Social views on psychiatric patients and psychiatry in the context of stigmatization]. PMID- 15825228 TI - [Nikolai Mikhailovich Itsenko (the 115th anniversary)]. PMID- 15825229 TI - Support the Ryan White CARE Act. PMID- 15825230 TI - Dating and daring to love again. One woman's journey after becoming a young widow. PMID- 15825231 TI - Model of neural circuit comparing static and adaptive synapses. AB - Replacing static synapses with the adaptive ones can affect the behaviour of neuronal network. Several network setups containing synapses modelled by alpha functions, called here static synapses, are compared with corresponding setups containing more complex, dynamic synapses. The dynamic synapses have four state variables and the time constants are of different orders of magnitude. Response of the network to modelled stimulations was studied together with effects of neuronal interconnectivity, the axonal delays and the proportion of excitatory and inhibitory neurons on the network output. Dependency of synaptic strength on synaptic activity was also studied. We found that dynamic synapses enable network to exhibit broader spectrum of responses to given input and they make the network more sensitive to changes of network parameters. As a step towards memory modelling, retention of input sequences in the network with static and dynamic synapses was studied. The network with dynamic synapses was found to be more flexible in reducing the interference between adjacent inputs in comparison to the network containing static synapses. PMID- 15825232 TI - Intellectual disability and psychotic disorders of adult epilepsy. AB - PURPOSE: To investigate the prevalence, psychopathology, and cognitive functions associated with psychotic disorders among adult epilepsy patients with intellectual disability (ID) based on a multicenter study in Japan. METHODS: The study was divided into three phases: a prevalence study of psychotic disorders among new referrals of epilepsy, a polydiagnostic comparative study of patients with psychotic epilepsy and those with schizophrenia, and a neuropsychological study of patients with psychotic epilepsy and education level-matched controls. RESULTS: Among 336 new referrals of epilepsy, a higher prevalence of psychotic disorders was found among patients with ID (24%) than among those with normal intelligence (6%). The psychotic symptoms and operational diagnoses of psychotic epilepsy patients with ID were similar to those of patients with normal intelligence. A wide range (7-86%) of psychotic epilepsy patients was diagnosed as having schizophrenia, depending on the operational criteria used. Patients with psychotic epilepsy had more disturbances in verbal memory and attention functions than controls. CONCLUSIONS: Epilepsy patients with ID show a predisposition to develop psychotic disorders. Distinguishing their psychotic symptoms from those of schizophrenia is difficult. Subtle cognitive disturbances predispose to psychotic disorders in epilepsy. PMID- 15825233 TI - What we confront with employment of people with epilepsy in Korea. AB - PURPOSE: To investigate the current status of employability in people with epilepsy (PWE) in Korea. METHODS: Data were collected from 543 adult epilepsy patients. Housewives and students were excluded. Information was obtained from self-completed questionnaires. RESULTS: The unemployment rate for Korean PWE was as high as 31%. This figure was five times higher than that of general population. The unemployed PWE had significantly lower quality of life than the employed PWE. the employability of PWE was significantly affected by certain factors: the frequency and severity of seizures; age at onset; interseizure psychosocial disabilities including self-esteem, personality, and problem-solving style; and social discrimination. Actual discriminatory practices in the employment of the PWE were prevalent in Korea. Overall, 24.5% of respondents believed that they had even been treated unfairly at work or when trying to get a job. More than half of those who disclosed their disease to employers reported that they have been refused a job because of epilepsy. About 75% of the patients surveyed said that they have never disclosed their disease in applying for a job. CONCLUSIONS: In Korea, PWE have faced many difficulties in employment. The discrimination against PWE and negative social attitudes toward epilepsy may play an important role in the employability of PWE. We need active participation in the "Out of the Shadows" world campaign. PMID- 15825234 TI - Current concepts in screening for noncommunicable disease: World Health Organization Consultation Group Report on methodology of noncommunicable disease screening. AB - Screening people for serious diseases is receiving increasing attention as studies demonstrates the potential benefits of early detection and early intervention in preventing morbidity and mortality. Screening tests are available for some of the most important noncommunicable diseases, including cardiovascular disease, type 2 diabetes and several site-specific cancers. However, screening guidelines for cardiovascular disease, cancers and diabetes vary within and between countries because many national and international organizations are developing their own guidelines. There is a need to address the issue of screening for disease, with an emphasis on providing accurate and practical information relevant to all countries. This report arose from a Consultation Group meeting held at WHO headquarters in Geneva. It presents an examination of the principles of screening with an emphasis on noncommunicable disease prevention and control policy. PMID- 15825235 TI - General discussion. PMID- 15825236 TI - Proceedings of a symposium on the relations between IgA nephropathy and tonsils. Okayama, Japan, April 13, 2003. PMID- 15825237 TI - Surgeon skill influences OPCAB success. PMID- 15825238 TI - The complexities of the chimeric practice model. PMID- 15825239 TI - [U.S. Army Dental Corps in World War II]. AB - The newly created U.S. Army Dental Corps, had to deal with a huge recruitment of troops after the Japanese attack of Pearl Harbour.Initially quickly overtaken by events, they gradually made up their lack of staff by the massive training of army dentists, in close co-operation with the American dental schools and by taking on many civil dentists. The lack of dental equipment and material was made up by the increased national output. With these advantages, both the mobile unites at the front and the dental clinics in the rear guard, appointed to the many regiments of differing size, proved to be well organised and successful. PMID- 15825240 TI - Evaluation of skilled leg movements in patients with stroke using a computerized motor-skill analyzer for the lower limb. AB - The utility of a new computerized motor-skill analyzer for the lower limb was studied in healthy subjects and patients with stroke. The computerized motor skill analyzer for the lower limb evaluated accuracy in a figure-tracking task with the foot. The intraclass correlation coefficients of measurements in subjects were .61 to .89 (p < .01). The trajectory errors of the unaffected lower limb in stroke patients were negatively correlated with physical ability. Trajectory errors of the patients were significantly larger than those of the healthy subjects. These results suggest that the computerized motor-skill analyzer is useful for quantitatively evaluating motor-skills of the lower limb. PMID- 15825241 TI - [Diagnosis and treatment of neoplasms in children. Proceedings of the 2nd Congress of the Polish Society of Pediatric Oncology and Hematology. 29-30 May 2003, Krakow, Poland]. PMID- 15825242 TI - Effects of Korean traditional herbal remedy on heart rate variability: linear and nonlinear analysis. AB - Heart rate variability was compared in 20 subjects taking traditional herbal remedy (Ondamtanggamibang) or placebo control pill, in placebo controlled design experiment. Herbal remedy group reported more pleasant and clam emotions than did the placebo group. Herbal remedy reduced the heart rate and increased heart rate variability (HRV) as indicated by a reduced low frequency/high frequency power ratio of heart rate variability. With nonlinear analysis, the Poincare plot index of HRV and approximate entropy was greater in the herbal remedy group than in the control group. These findings suggest that herbal remedy stabilizes the sympathovagal function and cardiac autonomic nervous system by inducing more positive emotions than does placebo therapy. In conclusion, herbal remedy may act by stabilizing both the autonomic nervous system and the emotional state. PMID- 15825243 TI - Non sinit esse feros. An essay on Ovid and the motto of the Royal College of Physicians of Edinburgh. AB - Questions are not infrequently asked about the meaning of the motto displayed on the College Arms. When a translation is offered--usually along the lines of 'It is forbidden to be cruel'--there is often puzzlement about why a College of Physicians should have chosen such a phrase to embody its aims and aspirations. In this essay I discuss the motto's relations to the couplet of Latin verse from which it derives and offer a possible explanation of how the misquotation--for such it is--that forms the motto may have come about. Then I say a little about the circumstances under which, some 2000 years ago, the Roman poet Ovid composed the poem which is its source. Finally, I show how consideration of an accurate text of Ovid's whole original couplet offers an interpretation of the motto that was apposite at the time of the College's foundation and is no less so more than three centuries later. PMID- 15825244 TI - Nerve, Muscle, and Beyond: the R.B. Stein Symposium. January 29 - February 1, 2004 in Vernon, British Columbia, Canada. PMID- 15825245 TI - Adolf Hitler's medical care. AB - For the last nine years of his life Adolf Hitler, a lifelong hypochondriac had as his physician Dr Theodor Morell. Hitler's mood swings, Parkinson's disease, gastro-intestinal symptoms, skin problems and steady decline until his suicide in 1945 are documented by reliable observers and historians, and in Morell's diaries. The bizarre and unorthodox medications given to Hitler, often for undisclosed reasons, include topical cocaine, injected amphetamines, glucose, testosterone, estradiol, and corticosteroids. In addition, he was given a preparation made from a gun cleaner, a compound of strychnine and atropine, an extract of seminal vesicles, and numerous vitamins and 'tonics'. It seems possible that some of Hitler's behaviour, illnesses and suffering can be attributed to his medical care. Whether he blindly accepted such unorthodox medications or demanded them is unclear. PMID- 15825246 TI - Screening for sexually transmitted diseases and hepatitis in 18-29-year-old men recently released from prison: feasibility and acceptability. AB - Men entering prisons have high rates of sexually transmitted disease (STD), hepatitis, and HIV. This study sought to determine the acceptability and feasibility of screening for STD and hepatitis in young men released from prison. Participants were interviewed six months after release and offered free screening. Of 42 (56%) eligible men who participated in the qualitative interview, 33 (79%) provided at least a blood or urine specimen. Eight of 33 (24%) men tested had chlamydia, trichomoniasis, hepatitis B or C virus (HBV or HCV). Three of 32 (9%) had chlamydia, three of 32 (9%) had trichomoniasis, two of 28 (7%) had prior syphilis, and two of 28 (7%) had HCV. Of 28 tested for HBV, six (21%) were immune, two (7%) had chronic infection, and 20 (71%) were susceptible. Barriers to screening included lack of forewarning, inconvenience, and insufficient incentive. In conclusion, screening for STD and hepatitis among former inmates can be acceptable and feasible. Forewarning, reducing the time burden, and providing monetary incentives may increase screening rates. PMID- 15825248 TI - Morphology 2003. Abstracts of the 41st Symposium of the Czech Anatomical Society with International Participation. September 15-16, 2003, Hradec Kralove. PMID- 15825247 TI - Prenatal maternal cortisol, fetal activity and growth. AB - Pregnant women (N = 131) were recruited at a prenatal ultrasound clinic between 20 and 28 weeks (M = 24 weeks) gestation. The women were given the CES-D for depression and the State-Trait Anxiety Inventory and were asked to provide a urine sample to be assayed for cortisol, norepinephrine, epinephrine, dopamine, and serotonin. Ultrasound sessions were conducted and coded for fetal activity and estimated fetal weight. Regression analyses were then conducted with fetal activity and fetal weight as outcome variables. Gestational age entered both analyses as the first variable followed by: (1) prenatal cortisol as a significant predictor of fetal activity; and (2) prenatal cortisol as a significant predictor of estimated fetal weight. PMID- 15825249 TI - Curare: the South American arrow poison. AB - The history of curare is both curious and convoluted. A product of South American culture it emerged in the sixteenth century from the mists of antiquity at the same time as quinine, coca, and chocolate. Like quinine, at first came the extract but no plant, and later the plant but no chemical compound. It took more than 300 years and the efforts of many explorers and scientists to resolve the problem. These included Condamine, Humboldt, Brodie, Waterton, Bernard, Dale, Walker, and King. Finally, the pure compound d-tubocurarine was isolated from the liana Chondrodendron and synthesised. Its specific physiological action was blockade of the effect of acetylcholine at the neuro-muscular junction. Such a paralytic poison could be used to kill oneself or others. The bizarre plot to kill the Prime Minister, Lloyd George, during the First World War is described. Fortunately this nefarious plan was thwarted by the Secret Service! PMID- 15825250 TI - Development of hepatotoxicity in HIV patients switching at least one protease inhibitor. AB - In order to evaluate the occurrence of hepatotoxicity in patients treated with antiretroviral therapy (ART) who switch protease inhibitor (PI), and the role of viral hepatitis in its development, we performed a retrospective study on 182 HIV patients treated with ART for 24 months. The presence of hepatitis viruses and alanine transaminase levels were evaluated. Hepatotoxicity developed in a low number of subjects without co-infection, but was significantly higher in co infected patients (14/51 versus 62/131, P = 0.01). Ritonavir was associated with higher rates of severe hepatotoxicity in the co-infected group. Patients presenting any problems related to ART, including the development of hepatotoxicity, continued therapy by switching PI. The occurrence of hepatotoxicity with second/third choice PIs, including ritonavir, remained stable. Our results suggest that switching PI does not increase the occurrence of drug-related liver toxicity. PMID- 15825251 TI - Complexity analysis of dense array EEG signal reveals sex difference. AB - This article deals with the complexity aspects of the recorded electroencephalogram (EEG) signal from male and female subjects. The analysis follows direct application of time series measures of global linear complexity and characterization of the embedded complexity in the signals using the nonlinear statistic of approximate entropy. The study reveals significant differences in complexity between the two sex groups during passive, no-task conditions, whereas no apparent variation exists during a mental task state. The detection of subtle changes as well as the ease in presenting a global picture of the complexity variation on the human cortical surface makes the nonlinear statistic a better marker of system complexity. PMID- 15825253 TI - Employee manuals. PMID- 15825252 TI - Genetic variations in vascular endothelial growth factor and endothelial nitric oxide synthase and their contributions to human disease. AB - This review addresses a rapidly growing area of vascular biology, i.e. genomic variations in vascular genes that underlie different human phenotypes. Two of the most important molecular in vascular biology, endothelial nitric oxide synthase (eNOS) and vascular endothelial cell growth factor (VEGF)are discussed. Variations in the eNOS gene have been correlated with a number of human diseases including hypertension, coronary vasospasm, smoking dependent risk of coronary disease, myocardial infarction and placental disruption. Similarly, variations in the VEGF gene have been associated with increased risk of various cancers, DiGeorge syndrome, psoriasis, diabetic renal disease and amyotropic lateral sclerosis. Understanding the molecular basis of these genetic variations and how they contribute to the pathophysiology provides new and important insights into human disease. PMID- 15825254 TI - [Social Psychiatry at the Charite. Proceedings of the symposium to commemorate the 30th anniversary of social psychiatry. Schizophrenia: genetics, cognition, therapy, rehabilitation. 11-12 June 2004, Berlin, Germany]. PMID- 15825255 TI - Space life sciences: life support systems and biological systems under influence of physical factors. PMID- 15825256 TI - Dermatology Update: Focus on Skin and Soft Tissue Infections. Chicago, Illinois, USA, 2003. Conference proceedings. PMID- 15825257 TI - Effects of air velocity on photosynthesis of plant canopies under elevated CO2 levels in a plant culture system. AB - To obtain basic data for adequate air circulation for promoting plant growth in closed plant production modules in bioregenerative life support systems in space, effects of air velocities ranging from 0.1 to 0.8 m s-1 on photosynthesis in tomato seedlings canopies were investigated under atmospheric CO2 concentrations of 0.4 and 0.8 mmol mol-1. The canopy of tomato seedlings on a plug tray (0.4 x 0.4 m2) was set in a wind-tunnel-type chamber (0.6 x 0.4 x 0.3 m3) installed in a semi-closed-type assimilation chamber (0.9 x 0.5 x 0.4 m3). The net photosynthetic rate in the plant canopy was determined with the differences in CO2 concentrations between the inlet and outlet of the assimilation chamber multiplied by the volumetric air exchange rate of the chamber. Photosynthetic photon flux (PPF) on the plant canopy was kept at 0.25 mmol m-2 s-1, air temperature at 23 degrees C and relative humidity at 55%. The leaf area indices (LAIs) of the plant canopies were 0.6-2.5 and plant heights were 0.05-0.2 m. The net photosynthetic rate of the plant canopy increased with increasing air velocities inside plant canopies and saturated at 0.2 m s-1. The net photosynthetic rate at the air velocity of 0.4 m s-1 was 1.3 times that at 0.1 m s-1 under CO2 concentrations of 0.4 and 0.8 mmol mol-1. The net photosynthetic rate under CO2 concentrations of 0.8 mmol mol-1 was 1.2 times that under 0.4 mmol mol-1 at the air velocity ranging from 0.1 to 0.8 m s-1. The results confirmed the importance of controlling air movement for enhancing the canopy photosynthesis under an elevated CO2 level as well as under a normal CO2 level in the closed plant production modules. PMID- 15825258 TI - [Abstracts of the 109th Annual Meeting of the Japanese Ophthalmological Society. 24-25 March 2005]. PMID- 15825259 TI - Pharmacokinetic, pharmacodynamic and pharmacogenomic approaches for measuring and managing variability in drug response. Abstracts of the 15th Keio University International Symposium for Life Sciences and Medicine. January 25-26, 2005, Tokyo, Japan. PMID- 15825260 TI - Abstracts of the 30th Annual Meeting of the German Society of Cell Biology. 16-19 March 2005, Heidelberg, Germany. PMID- 15825261 TI - Somatostatin receptor physiology and targets for somatostatin analogue therapy. Abstracts of the Young Investigators' Meeting. 28-30 January 2005, Barcelona, Spain. PMID- 15825262 TI - Abstracts of the 37th Congress of the Japan Epilepsy Society. Sendai, Japan, October 30-31, 2003. PMID- 15825263 TI - California ramps up stem cell plans. PMID- 15825264 TI - New studies raise global warming fears. PMID- 15825265 TI - [Health General, leader of private hospitalization in Europe]. PMID- 15825266 TI - Not so good vibrations. PMID- 15825267 TI - Howard Berg. PMID- 15825268 TI - Extracellular proteases: biological and behavioral roles in the mammalian central nervous system. AB - Extracellular proteases and their inhibitors have been implicated in both physiological and pathological states in the central nervous system (CNS). Given the presence of several classes of proteases, it is believed that each enzyme may undertake distinct biological roles. Some are indispensible for neuronal migration, neurite outgrowth and pathfinding, and synaptic plasticity. Others are required for neuronal death and tumor growth and invasion. Furthermore, studies from transgenic animals lacking or overexpressing one or more of the proteases have suggested that functional compensations and redundance among different members do exist. Normally, protease activity is tightly regulated by specific inhibitors to prevent disastrous proteolysis. Various insults can disrupt the fine control of proteolysis and caise pathological changes. Novel strategies have been attempted to maintain or restore protease-inhibitors homeostasis, thus minimizing damages to the CNS. They may provide us with effective therapeutic tools for fighting certain neurological disorders. PMID- 15825269 TI - [Experimental research on laser-induced single-molecule detection]. AB - Single-molecule detection is a kind of ultra-high sensitive detection technique. It is widely used in biology, medicine, environment and so on. Here we report briefly the principle and device of our laser-induced single-molecule detection spectrum instrument. We also report the experimental results on the instrument by using C445 dye, including the affection of sample velocity, energy characteristic, consistence linearity and detection limit. We discuss the effect of photon bleach on experimental results. These results are the base of our future work. PMID- 15825270 TI - [Spectral properties and energy transfer of Ce3+ and Eu2+ ions in Ca8Mg(SiO4)4Cl2]. AB - Ce3+ and Eu2+ codoped calcium magnesium chlorosilicate Ca8Mg(SiO4)4Cl2 (CMSC) was synthesized for the first time. The diffuse reflection spectra,excitation and emission spectra of CMSC : Ce3+, Eu2+ were measured and studied. Due to the good overlap between the Ce3+ emission bands and Eu2+ excitation bands, the energy transfer between Ce3+ and Eu2+ ions is expected to be very efficient and is confirmed by the emission spectra of CMSC : Ce3+, Eu2+ system. The fluoresence lifetime of Ce3+ in CMSC : 0.006Ce3+ and CMSC : 0.006Ce3+, 0.03Eu2+ was determined to be 30.1 and 17.2ns,respectively. By the calculation,the probability of energy transfer from Ce3+ to Eu2+ (PSA) is 2.49 x 10(7)/s and energy transfer efficiency (eta(SA)) is 42.9%. PMID- 15825271 TI - [Real-time remote detection of weak-spectra of chemical vapors using subtractive spectroscopy techniques]. AB - Infrared spectra from the several typical chemical vapors were observed remotely using the RFX-40 passive Fourier transform infrared spectrometer. In this paper, we discuss mainly the characteristics of infrared radiance transfer of chemical vapors. Subtractive spectroscopy was used to reduce the effect of atmosphere radiance on weak spectrum and enhance the spectral features of the chemical vapors. PMID- 15825272 TI - [Application of computer simulation in the study of vibrational distributions for the products from energy transfer reaction]. AB - The method of computer simulation in the study of vibrational distributions for the products from the energy transfer reaction is discussed. Taking the Ar (3P(0.2)) + PCl --> PCl (A3II) reaction as an example, we analysed the process of the simulation and the problems that should be paid attention to. The Franck Condon factors of PCl (A3II --> X3sigma(-1)) are reported for the first time. PMID- 15825273 TI - [The observations of plasma of AC operation in the CT-6B Tokamak by Ha emissions]. AB - The spatial distribution and temporal evolution of plasma of AC operation in the CT-6B Tokamak has been observed through Ha emissions by an optical multichannel diagnostic system for the first time. The results show the plasma density is not completely lost during current reversal, therefore, there were no need for second breakdown after the current reversal. The spatial and temporal evolution of plasma is dependent on the current wave forms. PMID- 15825274 TI - [Characteristics of the absorption spectra of the mixtures of C42(Al), C32 (Si) and so on]. AB - The mixtures containing C42 (A1), C32 (Si), C30 (Ca) and C28 (Fe) are produced by means of arc discharge and He gas convection. The spectra are measured and compared with the absorption spectra of all carbon molecules. The result shows that after imbeded in all carbon molecules, the Al, Si, Ca and Fe atoms do not change the positions of the absorption peak of original molecules, but only change the probability of pi --> pi* transition and n --> pi* transition of these molecules. PMID- 15825275 TI - [Enhancement of the resolution of overlapped spectra by derivative deconvolution]. AB - The principle of derivative deconvolution is introduced in this paper. By derivative deconvolution operated on simulated and measured spectra, the half width of peaks has been decreased, the resolution of overlapped spectra has been enhanced and therefore the accuracy of analytical study has been improved. PMID- 15825276 TI - [Study on the oxidation of N-alkylacridans and 10,10'-dialkyl-9,9'-biacridens by spectrophotometer]. AB - The oxidation of N-alkylacridans and 10,10'-dialkyl-9,9'-biacridens with pi acceptors TCNQ, DDQ, TCNE and CA were studied by UV, IR, ESR and fluorescent spectra, the oxidation mechanism for N-alkylacridans and 10,10'-dialkyl-9,9' biacridens was presented respectively. PMID- 15825277 TI - [Photothermal deflection spectroscopy for the detection of fluoride ion content in water]. AB - In this paper the application of photothermal deflection spectroscopy to the detection of fluoride ion content in water is described. This PDS method gets rid of two factors that impair the test sensitivity of regular photometric method applied to the measurement of solution concentration, and increases the sensitivity. That the PDS signal varied with modulation frequencies, pump laser powers and probe laser powers is discussed, and the appropriate parameters of test apparatus are chosen. PMID- 15825278 TI - [Detection of radical anion of nitrobenzene in situ on palladium electrode in different solvent]. AB - By means of a highly sensitive electrolytic cell installed in the ESR cavity, radical anion of nitrobenzene in the process of nitrobenzene electroreduction on palladium electrode in different solvent is dectected by flowing electrolytic system. The results show that in different solvent the discrimination of the ESR spectra are different, the solvent has an effect on nitrobenzene radical anion. The nitrobenzene electroreduction on Pd electrode must produce radical anion as an intermediate. PMID- 15825279 TI - [Library search of UV spectra of organic environmental pollutants by artificial neural network]. AB - In this paper, the artificial neural network (ANN) was applied to the library search of UV spectra of organic environmental pollutants. The optimization of network parameters was discussed and the effects of noise and impurity were investigated in detail. The use of derivative spectra for ANN library search was proposed to enhance the resolution of the UV spectra. This method could speed up the convergence of the network and could enhance the tolerance of impurity level, but was subject to noise comparing with the ANN using conventional UV spectra. Results show that the ANN is superior to the correlation coefficient method in resistance to noise and impurity. PMID- 15825280 TI - [Correction of spectral interferences by ICP-AES primary expert system]. AB - A subpackage for spectral interference correction has been set up in ICP-AES primary expert system. Some numerical recipes have been applied in this subpackage, such as curve smoothing, derivative spectrum, FFT technique, spectral resolving etc. A comprehensive method for spectral interference correction is presented. All these numeric recipes have been tested with simulated data and also utilized in several typical examples of real spectra. Results are satisfactory. PMID- 15825281 TI - [Spectra analysis of ignition flame in two-stroke gasoline engine burning blended fuel]. AB - In a two-stroke gasoline engine, exhaust gas pollution is especially severe when it burns rich mixture of blended fuel. The results of spectra analysis of ignition combustion flame show that the peak luminous intensities of three characteristic spectra CH(431.5nm), C2(516.5nm) and CN(387nm) are strong, of which the peak luminous intensity of CN(387nm) is the most outstanding. The mechanism of NO(x) formation in exhaust gas can be illustrated by Fenimore theorem. The mechanism was also verified by experiments conducted by Japanese researcher K. Nagase. PMID- 15825282 TI - [Commentary on the methods for studying the mechanism of electrothermal atomic absorption spectrometry]. AB - The methods for studing the mechanism of electrothermal atomic absorption spectrometry are critically reviewed 40 references and cited. PMID- 15825283 TI - [Determination of selenium by platform graphite furnace atomic absorption spectrometry]. AB - The matrix modifier and its amount, the ashing or atomization temperature of selenium and interference experiment are investigated. The experimental result shows that the application of Pd-Ca as mixed matrix modifier not only reduces interference but also enhances ashing temperature and increases sensitivity. The method can be used directly for the determination of selenium in cereal by platform graphite furnace atomic absorption. The characteristic mass is 39 ng,the detection limit is 0.11 microg/g, the relative standard deviation is 4.9-7.7% and the recovery is 92-100%. PMID- 15825284 TI - [Determination of germanium and selenium in malt powder by GFAAS]. AB - A method for determination of germanium and selenium in malt powder has been developed by graphite furnace atomic absorption spectrometry with L'vov platform. While (Pd+Ni) and Pd were used as matrix modifiers,the ashing temperatures of Ge and Se were increased to 1400 and 1200 degrees C respectively,and the matrix inferences were removed. The characteristic masses are 31 pg Ge and 23pg Se(0.0044 A), the detection limits are 28pg Ge and 62pg Se (3 sigma), the precisions (n = 9) are 3. 7-5.6%(Ge) and 4.3-6.5% (Se) and the recoveries are among 90-105%. PMID- 15825285 TI - [Indirect determination of pyritic sulfur in coal by flame atomic absorption spectrometry]. AB - An atomic absorption spectrophotometric method for the determination of pyritic sulfur in coal was developed based on oxidizing pyritic sulfur in coal into sulfate, which then reacted on an excess of solid barium chromate,and was followed by collection of the unreacted barium chromate and barium sulfate precipitates, and AAS determination of free chromate (equivalent to the sulfur originally present) at 357.9nm. The method was applied satisfactorily to the determination of pyritic sulfur in coal samples. The recovery of sulfur was 95 102% and relative standard deviation 3-5%. PMID- 15825286 TI - [Direct determination of zinc in polymerized rosin by flame atomic absorption spectrometry]. AB - Using 95% ethanol as the solvent, we directly determined zinc in polymerized rosin by flame atomic absorption spectrometry. The recovery rate is from 100 to 103 percent. The relative standard deviation (RSD) of the determination is between 1.1-4.1%. The sensitivity is high and the characteristic concentration reaches 0.0077 microg x mL(-1)/1%. The method is quite significant in forestry chemical works with its simplicity and efficiency. PMID- 15825287 TI - [Study of determination method for trace cobalt in serum of cancer patients]. AB - A method for the determination of micro amount of Co in serum of cancer patients by GFAAS has been established. The effect of protein in serum was eliminated by adding (1+1) HNO3. The blood serum was determined directly by adding Triton X-100 and NH4VO3 as matrix modifier. The method is handy and fast. The rate of recovery is 97.60-102.21% with RSD < 7.21. PMID- 15825288 TI - [Determination of trace selenium and tellurium in steel by hydride-generation nondispersive atomic fluorescence spectrometry]. AB - A method was developed for the determination of trace Se and Te in steel by ion exchange-separation hydride-generation nondispersive atomic fluorescence spectrometry. 40% (V/V) chlorhydric acid was selected as the best medium for hydride reaction. The detection limits of Se and Te are 0.27 and 0.33 ng/mL, relative standard deviations are 6.99 and 6.11%, respectively. The recovery was in the range of 96.7-102.0%. The results are satisfactory. PMID- 15825289 TI - [Column chromatographic preconcentration of trace copper in natural water using dithizone supported on naphthalene and determined by atomic absorption spectrometer]. AB - In this paper a column chromatographic preconcentration method using dithizone supported on naphthalene for copper in natural water is provided. The dissolving reagent is dimethylformamide (DMF). The dissolving solution is determined by atomic absorption spectrometer. The effect of pH, the flow rate of water samples, the choice of dissolving reagent, the effect of diverse ions were studied. RSD is 2.3%. The recoveries for the added standard are between 96%-103%. This method was applied to determined trace copper in natural water samples and standard water samples with satisfactory results. PMID- 15825290 TI - [Determination of total Ca in the lung tissue of rats by FAAS]. AB - Determination of total Ca in the Lung Tissue of Rats by FAAS was described, with a mixed microemulsion consisting of sodium dodecyl sulfate, N-dodecyl, N-dimethyl benzol ammonium bromide and nonyl phenol polyethyeneoxy ether. Comparing the result of the microemulsion FAAS with that of solution FAAS showed that the former was better than the latter in sensitivity, precision, selectivity and linearity of the calibration curve, with a detection limit of 1.92 microg x cm( 3), relative standard deviation of 1.92% (n = 10) and linear regressive coefficient of 0.9998 of the calibration curve over 0-20 microg x cm(-3) calcium. The method was applied to determine calcium amount in the lung tissue of rats, and the results were in accordance with those odtained by the conventional solution FAAS, in which a sample was reduced to ashes at a high temperature. PMID- 15825291 TI - [Burner head with high sensitivity in atomic absorption spectroscopy]. AB - This paper presents a burner head with gas-sample separate entrance and double access, which is used for atomic absorption spectroscopy. According to comparison and detection, the device can improve sensitivity by a factor of 1 to 5. In the meantime it has properties of high stability and resistance to interference. PMID- 15825292 TI - [Determination of the valence of iron by the conventional XRF spectrometer]. AB - FeKbeta emission spectra and parameters of mixtures are studied by conventional XRF spectrometer and PEAKFIT programs. The true spectra profile and it's parameters are obtained by profile resolving method. Integral area ratios are also calculated. The relations of these parameters and the valences of the iron are found. Fe3O4 sample and the iron ore which has different valences are quantified. PMID- 15825293 TI - [X-ray fluorescence analysis of Kaolin concentrate]. AB - An X-ray fluorescence technique was developed for the analysis of Al2O3, SiO2 and impurities in Kaolin concentrate. Two standard series, one for dual series of Al2O3-SiO2 and the other for all of the impurities, were prepared, respectively, from spectral pure agents based on the estimates of analyte content ranges and matrix effects in Kaolin concentrate. Powderd sample after determination of loss of ignition was pressed into disc by means of a little powderd sample preparation method modyfied in this paper. All analyte contents were calculated directly from the calibration curves without any matrix correction. In contrast to fusion technique the method has no the effects of flux impurities and dilute ration on the determination of minor and micro components in Kaolin concentrate. Errors of the results are < 1% for major component Al2O3, SiO2 and total, and < 5% for most and < 10% for few of the impurities. PMID- 15825294 TI - [Cyclodextrin induced-room temperature phosphorescence for the determination of quinoline without deoxidization]. AB - A cyclodextrininduced-room temperature phosphorescence (CD-RTP) system for the determination of quinoline without deoxidization procedure was developed based on the external heavy atom effect of dibromopropane (DBP), the RTP maximum wavelengths lambda(ex)/lambda(em) = 273/496nm. The difference of heavy atom effect between dibromopropane DBP and dibromoethane (DBE) was examined and the apparent formation constants of the ternary inclusion complexes for quinoline/beta-CD/DBP and quinoline/beta-CD/DBE systems were determined. As compared with DBE, the heavy atom effect of DBP was higher and the RTP intensity was stronger. The RTP intensity was linear for quinoline/beta-CD/DBP systems with quinoline concentration in the ranges of 5.0 x 10(-7)-5.0 x 10(-5)mol/L and 5.0 x 10(-5)-5.0 x 10(-4)mol/L, respectively. The detection limit was 7.8 x 10( 8)mol/L. The determination precision was satisfactory. PMID- 15825295 TI - [The fluorescence properties of DaYaWan soil humic acid]. AB - The fluorescence properties of humic acid derived from DaYaWan soil were analyzed using the conventional fluorescence and synchronous scan fluorescence spectroscopy. The results show that synchronous technique applied to the study of humic acid can reduce the fluorescence spectrum of a compound to a single sharp peak, and decrease the signal overlapping. In addition, the effects of various deltalambda, concentration and pH values on synchronous fluorescence spectroscopy were investigated in detail. PMID- 15825296 TI - [Study of the behavior on meso-Tetra (4-sulfophenyl) Porphyrin (TPPS) in the microemulsion system]. AB - The change of absorption spectrum of meso-Tetra (4-sulfophenyl) Porphyrin (TPPS) in microemulsion system (Triton X-100-nonane-pentanol-water) was studied. The electric dissociation constants (Ka ) and dimerization constants (KD) were calculated. It was shown that Ka of TPPS is added but KD of TPPS is decreased in microemulsion system as compared with that in aqueous solution. PMID- 15825297 TI - [Indirect determination of trace amount of oxalic acid radical by the I2-CCl4 extraction spectrophotometry]. AB - The article studies that in dilute H2SO4 dielectric substance IO3(-) reacts with C2O4(2-) to release certain amount of iodine under certain reaction condition: 2IO3(-) + 5C2O4(2-) + 12H+ = I2 + 10CO2 increasing + 6H2O therefore the method of indirect determination with I2-CCl4 extraction spectrophotometry is established. The reaction temperature is controlled in 90 degrees C; the reaction time 3 minutes; the reaction acidity pH1.0-2.0. The common ion basically does not affect the determination. The lowest detection limit of C2O4(2-) is 0.08 microg x mL( 1). After comparing the method of the recovery experiment with that of other determination, a satisfactory result is aquired and the recovery is between 96 98%. PMID- 15825298 TI - [Study on colour reaction of iron(II)-thibarbituric acid-nitrite system and its analytical application]. AB - In this paper, the optimum conditions of colour reaction of iron (II) thibarbituric acid-NO2(-) system, existent state and spectrophotometric characteristies of the complex were studied, and the reaction mechanism discussed. A new spectrophotometric method for the determination of trace iron was established. In a buffer solution of borate/NaOH of pH9.5, iron (II) and thibarbituric acid-NO2(-) react to form a stable blue complex anion, for which the maximum absorbance is at 645nm and the molar absorptivity is 2.66 x 10(4)L x mol(-1) x cm(-1). Beer's law is obeyed in the range of 0-40 microg/25mL for iron (II). The method is simple and rapid, shows satisfactory selectivity and precision and has been applied to determine iron in waters with satisfactory results. PMID- 15825300 TI - Detection of R factors in naturally occurring Vibrio anguillarum strains. AB - R factors were detected in Vibrio anguillarum strains from vibrio-diseased freshwater fish, ayu (Plecoglossus altivelis), in Japan. It was found that 65 out of the 68 random isolates from epidemics of vibrio disease in 1973 carried transferable drug resistance factors. The most common type determined resistance to sulfonamides, streptomycin, chloramphenicol, and tetracycline and belong to the fi(-) type. The high incidence of R factors in Vibrio anguillarum from cultured fish is assumed to be due to selective pressure exerted by chemotherapeutic agents used in fish culturing. PMID- 15825299 TI - Effect of amphotericin B and clotrimazole on lymphocyte stimulation. AB - Human lymphocytes were stimulated in vitro by phytohemagglutinin, concanavalin A, pokeweed mitogen, purified protein derivative-tuberculin, and allogenic cells. The deoxyribonucleic acid synthesis of the lymphocytes was inhibited in increasing degree by 4 to 8 mug of amphotericin B per ml of culture irrespective of lymphocyte stimulant used. The effect of clotrimazole on the deoxyribonucleic acid synthesis varied between experiments with different and also between experiments with the same stimulant. Ten micrograms of clotrimazole per ml was generally inhibiting, whereas in one experiment 2 mug or more per ml inhibited the purified protein derivative-induced deoxyribonucleic acid synthesis. The effects of amphotericin B and clotrimazole were neutralized by serum. PMID- 15825301 TI - Isolation and antibacterial activity of acylphloroglucinols from Myrtus communis. AB - Isolation procedures of two new acylphloroglucinols, myrtucommulone-A and myrtucommulone-B, from Myrtus communis leaves are given. Myrtucommulone-A was highly antibacterial against gram-positive bacteria but was not active against gram negatives. The chemical relation to other acylphloroglucinols and the antibacterial activity of the compounds isolated are discussed. PMID- 15825302 TI - In vitro susceptibility of Bacteroides corrodens and Eikenella corrodens to ten chemotherapeutic agents. AB - Forty strains of Bacteroides corrodens and Eikenella corrodens were tested to determine their degree of susceptibility to 10 commonly prescribed chemotherapeutic agents. Significant differences were evident between strictly anaerobic strains and facultatively anaerobic strains. PMID- 15825303 TI - Bacteriocin production and susceptibility among strains of Streptococcus mutans grown in the presence of sucrose. AB - Whereas the bacteriocin activity of eight strains of Streptococcus mutans was unaffected by growth in the presence of sucrose, such a medium rendered five S. mutans and one S. salivarius indicator strains unsusceptible to the bacteriocins. In contrast, the susceptibility of three S. faecalis and one S. pyogenes strains was unaltered when these indicators were grown in the presence of sucrose. Since the S. mutans and S. salivarius strains produced extracellular polysaccharides from sucrose whereas the S. faecalis and S. pyogenes strains did not, it was concluded that a coating of extracellular polysaccharides rendered normally susceptible organisms unsusceptible to bacteriocin action. This supports previous suggestions that such bacteriocins are not active in vivo and therefore play no role in regulating the microbial ecology in dental plaque. PMID- 15825304 TI - Evaluation of the efficacy of minocycline therapy for staphylococcal soft-tissue infection. AB - Ten patients with soft-tissue infections due to Staphylococcus aureus were treated with minocycline, a semisynthetic tetracycline with potent in vitro antistaphylococcal effects. Serum concentrations averaged three to five times the concentration of minocycline required to inhibit growth of S. aureus in vitro. Clearing of the infecting organism was slow (less than 50% of lesions were sterile on day 10 of therapy), but clinical improvement was noted in 8 of 10 patients. PMID- 15825305 TI - Empirical equation for pharmacokinetic analysis of drug serum levels after oral application. AB - The equation log(concentration) = a + b/ [Formula: see text] + c. [Formula: see text] + d.(T - phi) was used to calculate serum level curves from individual data sets of drug serum concentrations, obtained from experiments with orally administered drugs. These curves were subsequently used to calculate peak values, times for onset of peak, area, and other pharmacokinetic parameters that ought to be independent of any preconceived theory about the behavior of the drug in the system. A program is described by which parameters for individual data sets can be calculated and a mean curve with a peak value, time for onset of peak, and area equal to the arithmetic mean of the corresponding values for the participating subjects, is produced. The results of this method are compared to those of the "one-compartment model with lag time" and shown to be superior in all the test cases. In particular, the proposed method performs well with data from a highly protein-bound drug, for which the one-compartment model fails completely. Data sets of six to eight samples taken over the entire period of detectable serum levels, with 10% analytical error in the results, gave estimates of peak value and area with a coefficient of variation of 6 to 9%, whereas the variation in the estimates for different subjects in a treatment group amounted to 20 to 60%. This shows that the proposed method, although able to cope with a short series of imprecise measurements available in practical work, is still sufficiently sensitive to detect real differences between the individual subjects. PMID- 15825306 TI - Clinical pharmacology of pivampicillin. AB - Studies on pivampicillin hydrochloride and ampicillin trihydrate, administered in capsules to healthy volunteers, indicated that pivampicillin was absorbed more efficiently from the gastrointestinal tract than ampicillin. Average peak concentrations of ampicillin in the serum after doses equimolar to 250 mg of ampicillin were 6.8 mug/ml at 56 min with pivampicillin and 1.96 mug/ml at 1 h 24 min with ampicillin. The maximal concentration after pivampicillin treatment was also higher than that recorded when twice the equimolar dose of ampicillin, which averaged 3.2 mug/ml at 1 h 42 min, was used. The urinary excretion of ampicillin, expressed as a percentage of the administered dose, averaged 67 to 73 and 25 to 29% after administration of pivampicillin and ampicillin, respectively. The bioavailability of ampicillin, taken as the area under the serum curve, obtained with pivampicillin at a 250-mg ampicillin dose level was superior to that obtained with a 500-mg dose of ampicillin. Comparison of a suspension intended for children, containing the pivampicillin free base with a suspension of ampicillin trihydrate, emphasized the difference recorded for the capsule preparations. Administration of pivampicillin with a meal rich in fat and protein had no depressant effect on the absorption. Concurrent administration of probenecid caused higher and prolonged concentrations of ampicillin in the serum. PMID- 15825307 TI - Effect of the systemic fungicide carboxin on electron transport function in membranes of Micrococcus denitrificans. AB - The systemic fungicide carboxin (5,6-dihydro-2-methyl-1,4-oxathiin-3 carboxanilide) inhibited oxidation of succinate by membranes prepared from Micrococcus denitrificans, the K(i) being 16 muM. Oxycarboxin (5,6-dihydro-2 methyl-1,4-oxathiin-3-carboxanilide-4,4-dioxide), F831 (5,6-dihydro-2-methyl-1,4 oxathiin-3-carboxanilide-4-oxide), and another succinate oxidase inhibitor, 4,4,4 trifluoro-1-(2-thienyl)-1,3-butanedione (TTB) were less effective inhibitors of succinate oxidation by membranes of M. denitrificans. Oxidation of other substrates (nicotinamide adenine dinucleotide, reduced form, d-lactate, l lactate, malate, and d,l-alpha-hydroxybutyrate) was inhibited to a lesser degree by carboxin, and formate oxidation was entirely resistant. With all substrates tested, oxycarboxin, the dioxide analogue of carboxin, was less effective than carboxin. Carboxin also inhibited dichlorophenol indophenol (DCIP) reductase activities by these membranes in a manner both qualitatively and quantitatively similar to the inhibition of oxidation of the various substrates. The inhibition of DCIP reductase activities by TTB was qualitatively similar to carboxin, but TTB was a less effective inhibitor with all substrates tested. The inhibition of DCIP reductase by carboxin could be relieved by phenazine methosulfate with all substrates except d-lactate. Only slight inhibition of d-lactate-stimulated uptake of [(14)C]glycine by these membrane vesicles was seen with carboxin. Uptake of [(14)C]glycine could be stimulated to varying degrees with the other substrates tested, but in no case did carboxin cause significant inhibition. Membranes isolated from M. denitrificans are a useful system for investigating the mechanism of inhibition of electron transport function by carboxin, and the use of this system for evaluations of carboxin and its metabolites is suggested. PMID- 15825308 TI - Biological activity of antibiotic G-418, a new micromonospora-produced aminoglycoside with activity against protozoa and helminths. AB - On the basis of parallel in vitro studies with antibiotic G-418, gentamicin, neomycin, and kanamycin, antibiotic G-418 was found to be less potent than gentamicin but more active than either kanamycin or neomycin against most strains, with the exception of Pseudomonas, for which neomycin was more active than antibiotic G-418, and enterococci, for which antibiotic G-418 was more active than the other three antibiotics. Mouse protection tests indicated that antibiotic G-418 is approximately half as potent as gentamicin and its acute toxicity is one-half to one-third that of gentamicin. PMID- 15825309 TI - Interaction of clindamycin and gentamicin in vitro. AB - The minimal inhibitory concentrations of clindamycin and gentamicin alone and in combinations were determined by a microdilution method for 163 aerobic, facultative, and anaerobic clinical isolates. All 77 strains of Staphylococcus aureus, Diplococcus pneumoniae, Streptococcus pyogenes, and anaerobic bacteria (except for three strains of Clostridium) were inhibited by 1.6 mug or less of clindamycin per ml. Gentamicin did not interfere with the activity of clindamycin within the range of concentrations tested (0.1 to 100 mug/ml); for some strains combinations were synergistic. Sixty-two (94%) of 66 strains of Enterobacteriaceae and Pseudomonas aeruginosa were inhibited by 6.2 mug or less of gentamicin per ml. Combinations of clindamycin and gentamicin were indifferent for 29 strains and synergistic for 33 strains. All 20 strains of enterococcus, three strains of Clostridium, three strains of Escherichia coli, and one strain of Proteus rettgeri were resistant to both clindamycin (minimal inhibitory concentration greater than 3.1 mug/ml) and gentamicin (minimal inhibitory concentration greater than 6.2 mug/ml). Combinations of clindamycin and gentamicin were indifferent for 16 and synergistic for 11 of the resistant strains. Except for clindamycin-sensitive isolates, synergy was usually observed only at concentrations of one or both drugs which are not readily obtainable in vivo. Antagonism was never observed. PMID- 15825310 TI - Triple crossover study on absorption and excretion of ampicillin, pivampicillin, and amoxycillin. AB - The absorption and excretion of equivalent doses of ampicillin, pivampicillin, and amoxycillin were compared in 10 healthy volunteers after administration of these antibiotics in a randomized triple crossover fashion. As recommended, ampicillin and amoxycillin were administered on an empty stomach, whereas pivampicillin was given with breakfast. The mean half-life times were nearly identical for all three antibiotics: 60.9 min for ampicillin; 58.4 min for pivampicillin; and 62.2 min for amoxycillin. The absorption of the drugs was evaluated according to the following three criteria: the mean peak serum levels; the "area under the curve"; and the percentage of recovery in urine. By these criteria, pivampicillin was the best-absorbed drug, with absorption 3.0 to 3.6 times higher than that of ampicillin and 1.2 to 1.5 times higher than that of amoxycillin, whereas the absorption of amoxycillin was 2.1 to 2.9 times higher than that of ampicillin. PMID- 15825311 TI - Antivirion effects of streptovaricin complex against Friend virus. AB - The in vitro antivirion activities of five different streptovaricin complex lots against the polycythemic strain of the Friend virus were evaluated. The assay system was based on the inhibition of the Friend virus-induced spleen foci. The virus inactivation process was shown to be susceptible to variation in temperature, pH, and time. The antivirion activity and the acute toxicity for mice, as well as the optical properties of these streptovaricin complexes, do not co-vary; this suggests that their biological activities are not associated with a single molecular structure. In addition, the antivirion activity of the five preparations of streptovaricin complex differs about 30-fold, indicating that this activity does not reside in a major component of the complex. PMID- 15825312 TI - Assessment of cytosine arabinoside as an antiviral agent in humans. AB - Cytotoxicity, minimal inhibitory concentrations of herpesviruses, and pharmacokinetic studies of cytosine arabinoside (ara-C) were done. Ara-C compared favorably with idoxuridine in in vitro studies of antiviral activity versus herpes simplex, varicella-zoster, and cytomegalovirus. However, ara-C was 10 times more toxic to tissue cultures, and concentrations in serum and urine of three patients who were given ara-C at acceptable dosages (1 mg/kg per day) were not measurable by our assay. These studies predict that ara-C is not likely to be a useful antiviral agent in humans because its therapeutic to toxic ratio approaches unity. These predictions of little clinical efficacy seem now to have been confirmed by clinical trails in humans. Pharmacokinetic studies outlined here should precede and help formulate controlled clinical trials of potential antiviral agents in humans. PMID- 15825313 TI - Paper disk-agar diffusion assay of penicillin in the presence of streptomycin. AB - Microbiological assay of individual antibiotics in mixtures of antibiotics depends on the use of selective inactivation and/or of test bacteria with differential susceptibility. Controlled experiments revealed that streptomycin in concentrations of 20 and 40 mug/ml did not influence a disk diffusion assay of penicillin with Sarcina lutea (ATCC 9341) as the test organism. In the case of penicillin concentrations less than or equal to 1 IU/ml, addition of 80 mug of streptomycin per ml influenced the penicillin assay significantly. Clinical use of streptomycin resulting in levels above 40 mug/ml usually did not occur; therefore penicillin could be assayed as though streptomycin were not present. We observed additionally that S. lutea was unable to grow on agar plates prepared with semicarbazide hydrochloride. PMID- 15825314 TI - Effect of leucomycins and analogues on binding [14C ]erythromycin to Escherichia coli ribosomes. AB - We examined the effect of leucomycins, leucomycin derivatives, and other 16 membered macrolides (tylosin, niddamycin, spiramycin I, and spiramycin III) on [(14)C]erythromycin binding to ribosomes. Results of these studies enabled determination of the association and dissociation constants for the binding of each of these compounds to Escherichia coli ribosomes. In addition, the binding of the leucomycins and the leucomycin derivatives to ribosomes in general correlated with their antimicrobial activity. PMID- 15825315 TI - Proflavine and light in the treatment of experimental herpetic ocular infections. AB - The effect of purified proflavine and light exposure was assessed in rabbits whose eyes had been infected with one of two strains of herpesvirus. In comparing proflavine-light and placebo-light treatment, 0.1% proflavine administered twice daily for 5 days had a significant effect in suppressing herpetic eye disease, but 0.05% proflavine was less effective. In addition to being effective in infections with either virus strain, the 0.1% proflavine also suppressed intensity of corneal epithelial ulceration and stromal opacity in animals pretreated with subconjunctival corticosteroids to produce more severe disease. Proflavine or idoxuridine (IDU) alone or in combination showed no differences in suppressing herpetic ocular disease, but all were significantly more effective than placebo. Virus recovery rates were approximately the same from eyes treated with proflavine, IDU, or placebo, indicating that viral replication in the cornea and conjunctiva was not completely suppressed by either of the antiviral drugs alone or in combination. PMID- 15825316 TI - Antimicrobial susceptibility of Haemophilus influenzae. AB - Selected clinical isolates of Haemophilus influenzae were tested for their susceptibility to seven antibiotics by a microtiter broth dilution and an agar diffusion method. Eleven of 40 strains tested were resistant to ampicillin, the drug of choice, by both methods. All the strains tested were susceptible to chloramphenicol, and all but one were susceptible to tetracycline. Of the other four antibiotics tested, the ranges of minimal inhibitory concentrations (MICs) were: 0.5 to 4 mug of gentamicin per ml, 0.5 to 4 mug of erythromycin per ml, and 2 to 16 mug of clindamycin per ml. MICs of ampicillin for both the susceptible and resistant strains were markedly affected by inoculum concentration. The ampicillin MICs of the resistant strains were also affected by the time of incubation. PMID- 15825317 TI - Beta-lactamase activity in ampicillin-resistant Haemophilus influenzae. AB - The specific activity, substrate profile, response to inhibitors, inducibility, and cellular localization of the beta-lactamase produced by an ampicillin resistant strain of Haemophilus influenzae type B were investigated. In these properties the enzyme resembles beta-lactamases produced by other gram-negative bacilli more closely than those produced by gram-positive organisms. It is quite similar to an enzyme found in strains of Klebsiella pneumoniae, and differs significantly from those described in other gram-negative bacilli. Comparison of the substrate profile with the minimal inhibitory concentrations of various beta lactamase antibiotics suggests that the beta-lactamase plays an important role in the antibiotic resistance of this organism. PMID- 15825318 TI - Effect of adenosine deaminase upon the antiviral activity in vitro of adenine arabinoside for vaccinia virus. AB - This study determined that the effect of 9-beta-d-arabinofuranosyl-adenine (adenine arabinoside, Ara-A) upon vaccinia virus plaque development in the stable monkey kidney line, LLC-MK(2), was increased approximately 40-fold when an inhibitor of adenosine deaminase (ADA) was added to the tissue culture media along with infective inocula. The concentration of Ara-A required to completely suppress plaque development (total plaque inhibitory concentration(100); TPIC(100)) was greater than 10 mug/ml. However, when ADA activity was inhibited, the TPIC(100) was 0.5 mug/ml or less. Chromatographic assay of arabinosylpurines in the media provided evidence that adenine arabinoside was rapidly deaminated to 9-beta-d-arabinofuranosylhypoxanthine by the cellular monolayers, in the absence of animal serum, and that the rate of deamination, at 5 mug/ml, by the cells was equal to the rate of diffusion of Ara-A across the cellular membrane. The half life of Ara-A in the media, starting with 5 mug/ml, was 2 to 3 h and shorter at lower concentrations. The study demonstrates the profound effect that an indicator system, acting as an intact biological unit, can have upon a potential antiviral compound. PMID- 15825319 TI - Antibacterial effects of silver electrodes with weak direct current. AB - Silver, platinum, gold, stainless-steel, and copper electrodes were used with low currents (0.02 to 20 muA/mm(2)) to explore their electrochemical effects on the growth of four bacterial species. In the higher current ranges, all electrodes inhibited growth at both poles, usually in conjunction with electrolytic break down of the medium and severe corrosion of the metal. Silver, however, was extremely bacteriostatic, even at the lowest current, when used as the anode. Quantitative studies showed that most of this inhibition takes place in a few hours and is not accompanied by changes in pH. Electrochemically injected silver from the anode is probably the instrumental agent, being effective in concentrations of about 5 mug/ml. This is the equivalent concentration of silver sulfadiazine that has been shown to give complete inhibition of bacteria, but without the sulfonamide moiety. PMID- 15825320 TI - Comparative in vitro evaluation of BL-P1654 and carbenicillin against Pseudomonas. AB - Prior investigators have reported a broad gram-negative spectrum and a marked anti-Pseudomonas activity in vitro with BL-P1654, 6-[R-alpha (guanylureido)phenylacetamido]-penicillanic acid. An in vitro comparison of BL P1654 to carbenicillin was performed against 84 strains of Pseudomonas and 39 strains of Enterobacteriaceae. Mean minimal inhibitory concentrations in Mueller Hinton broth of BL-P1654 and carbenicillin were 12.5 and 100 mug/ml, respectively. Bactericidal studies with representative strains of Pseudomonas showed a consistent lack of bactericidal activity with BL-P1654. Mean bactericidal concentrations of BL-P1654 and carbenicillin were >/=200 and 100 mug/ml, respectively. The lack of bactericidal activity of BL-P1654 was further substantiated with the "killing curve" technique. Based on these data, BL-P1654 can not be considered a substitute for carbenicillin as the penicillin analogue of choice for Pseudomonas infections. PMID- 15825321 TI - Synthesis of vaccinia virus polypeptides in the presence of hydroxyurea. AB - Hydroxyurea (5 x 10(-3) M) inhibits the development of vaccinia virus in HeLa cells early after infection and prevents the synthesis of viral deoxyribonucleic acid. In the presence of hydroxyurea, late viral polypeptides are not formed, as observed by electrophoresis on polyacrylamide gels. PMID- 15825322 TI - Microbiological assays and bioautography of maytansine and its homologues. AB - A quantitative microbiological assay and a bioautography system with Penicillium avellaneum UC-4376 was developed for the antitumor drugs maytansine, and its homologues, maytanprine and maytanbutine. The susceptibility of the assay is 1.5 mug/ml. PMID- 15825323 TI - Ampicillin resistance in Haemophilus influenzae as determined by a rapid test for beta-lactamase production. AB - A rapid capillary tube method was used to demonstrate beta-lactamase production by ampicillin-resistant strains of Haemophilus influenzae. PMID- 15825325 TI - [Time resolved photoluminescence of PPV derivatives/C60 combination system]. AB - We report the integrated and picosecond time-resolved photoluminescence (TRPL) measurement from two kinds of combination films: Poly(2-methoxy-5-(4 butenyloxy)phenylene vinylene) (MB-PPV)/C60 and Poly (2-methoxy-5-(4'-bromo butoxy) phenylene vinylene) (MBB-PPV)/C60. Comparing with the pure MBB-PPV film, PL weakening and quenching of MBB-PPV were observed in the multilayer and mixed MBB-PPV/C60 films respectively. From TRPL spectra, the change of PL decay lifetime could be clearly seen, those could be attributed to the excitation transfer (ET) process between the excited MBB-PPV molecule and C60 molecule. Further measurements indicate that there is no noticeable dependence of the ET process on the temperature in the combination films. PMID- 15825324 TI - Mutagenic activity of diazoacetylglycine derivatives. AB - Some diazoacetyl derivatives of glycine known as antitumor and immunosuppressive agents have been found to be powerful mutagens of the base substitution type. PMID- 15825326 TI - [Indirect sensitized upconversion in Tm3+ and Yb3+ codoped non-crystal pentaphosphate]. AB - We report the blue upconversion luminescence of Tm3+ and Yb3+ codoped non-crystal pentaphosphate pumped by -798nm laser diode. The samples were directly excited to 3F4 level of Tm3+ ion. Two energy transfer processes between Tm3+ and Yb3+ contribute mainly to the population of 1G4 level of Tm3+. Phonon plays an important role in the upconversion process. PMID- 15825327 TI - [The evaluation of parameters pertinent to the excitation in aurora with a time domain recursive instrumental-variables algorithm]. AB - The time-domain expression and spectral analysis of the combined direct and indirect excitation process in aurora are described. The technique to evaluate the parameters pertinent to the excitation in aurora with a time-domain recursive instrumental-variables algorithm was studied. The parameters thus determined compare favourably with those evaluated by means of a non-recursive cross spectral technique. A modification to the algorithm allows the tracking of changes in the excitation mechanism by evaluating parameters which vary in time. Processing of real auroral photometric measurements with this method has established the existence of such changes and demonstrated their time history. PMID- 15825328 TI - [Spectroscopic ellipsometry characteristics and the optical properties of liquid crystalline copolyesters]. AB - The optical properties of two thermotropic liquid crystalline copolyesters were characterized by spectroscopic ellipsometry and UV-visible spectroscopy. The differences in the refractive index variation and chromatic dispersion have been revealed and discussed in terms of differences in the vibrating transition of excited electrons and the molecular structures of the polymers. PMID- 15825329 TI - [Ultrasensitive analysis of R6G and R110 in solution]. AB - This paper describes the ultrasensitive detection of R6G and R110 in a flowing aqueous sample, based on laser-induced fluorescence (LIF) combined with mode locked pulsed excitation and time-correlated single-photon counting. A high repetition rate mode-locked argon ion laser operated at 514.5 nm was used as the excitation source. The laser beam was found tightly into a thin wall capillary flow cell. The probe volume was about 12.6 x 10(-12)L. Fluorescence was collected at 90 degrees using a x 40 microscope objective, and was detected by a fast response PMT detector. A time-gate was used to discriminate Raman scattering light background. Using the criterion of 3 standard deviations above background, the detection limits obtained for R6G and R110 were 3.8 x 10(-14) mol/L and 1 x 10(-13) mol/L respectively, and the average probability was lower than 1 in the presence of a R6G or a R110 molecule in the detection volume. PMID- 15825330 TI - [Recursion formula of energy levels for oxygen-like ions]. AB - A recursion formula of energy levels for Oxygen-like ions has been given based on the generalization of the perturbation expansion. It is found that the formula is considerably simple and accurate. PMID- 15825331 TI - [Analysis of the properties of plasma produced by pulsed laser ablation of metal Al at different ambient pressures]. AB - A time and space resolved diagnostic technique was used to study the properties of plasma produced by a pulsed laser on an Al target irradiated at different ambient pressures. The velocities of projected particles were measured. The projected ranges for 1-10keV AI2+ ions ejecting into ambient gas at different pressures were calculated in theory. The calculated results can explain the observed experimental phenomena successfully. PMID- 15825332 TI - [The thin film and absorption spectra of CsCu2I3]. AB - This paper introduces a method of manufacturing CsCu2I3 thin films through hot evaporation. According to the absorption spectra at low temperature, the excitonic coefficient of CsCu2I3 was calculated. The spectral analysis shows that the localization of electron and exciton excitations in the twofold Chain created by CuI4(3-) causes the complicated character of the optical spectra of CsCu2I3. PMID- 15825333 TI - [Study on the interaction of lauryl sodium sulfate and rhodamine-6G before micellar formation]. AB - The absorption spectra of an aqueous solution of Lauryl sodium sulfate (SDS) and Rhodamine-6G (R6G) over a wide concentration range were studied. The results show that when c(sds) < 1 x 10(-4)mol x L(-1) SDS forms complex with R6G with molar ratio of 1 : 1, but when c(sds) > 1 x 10(-4) mol x L(-1) the premicelle, which affects the aggregate state of R6G obviously, is formed in the solution. With SDS present, the aggregation number and aggregation constant of R6G were determined, and the dissociating of R6G by inorganic salts also investigated. PMID- 15825334 TI - [Study on the infrared spectra of 9-methoxy benzoxanthene-3,4-dicarboximide fluorescent dyes]. AB - The characteristic infrared spectra of eight novel 9-methoxy benzoxanthene-3, 4 dicarboximide fluorescent dyes was studied and the substiuent influences on the spectrum nature was also pointed out. The nature of N-substituent group in the imide ring affects the position of v(c=o) by mesomeric and inductive effects. Imides give the rise to two vc=o absorptions, namely v(a)(c=o) and v(as)(c=o) (1600-1700cm(-1)). The separations between v(a)(c=)o and v(as)(c=o) were about 36 43cm(-1). PMID- 15825335 TI - [Raman spectrum characteristics of methane]. AB - Two characteristic Raman peaks of methane at 2917 and 3020cm(-1) were measured. The results show that the four C-H bonds of methane molecular are unequal. The assignment of Raman bands at 2917 and 3020cm(-1) was discussed in this paper. PMID- 15825336 TI - [Effect of surface plasma resonance absorption on the intensities of Raman bonds with different frequency shifts]. AB - Obvious differences in intensities of the Raman bands with different frequency shifts were found for prydine molecules and their derivatives adsorbed on silver sol that were excited by different wavelength light. This phenomenon was analysed. PMID- 15825337 TI - [Research of eximer laser-excited fast speed spectrochemical analysis]. AB - Experimental method including XeCl eximer Laser-excited, optical multichannel analyzer (OMA) combined with the simultaneous spectrum widen technique was introduced in this paper. We programed data acquisition software and tested alloy steel sample. The acquisition result of 150nm simultaneous spectrum was showed. The charaterics and significances of our research were discussed. PMID- 15825338 TI - [Determination of impurities in tantalum by ICP-AES]. AB - The spectral and non-spectral inteferences of tantalum to the determination of Al, Ca, Cr, Cu, Fe, Mg, Mn, Mo, Nb, Ni, Pb, Sn and Ti by ICP-AES were studied. The non-spectral inteference can be eliminated by optimizing operating conditions. Three methods for correction of spectral inteferences were evaluated. The proposed method was used for the determination of impurities in synthetic samples with satisfactory results. PMID- 15825339 TI - [Direct collection with graphite probe filters and determination of trace cadmium in atmospheric particulate matter by graphite probe furnace atomic absorption spectrometry]. AB - After atmospheric particulate matter was directly collected with graphite probe filters, the trace cadmium in APM was determined by graphite probe furnace atomic absorption spectrometry. The results showed that the absorbance (peak-area mode)and the concentration of cadmium had a excellent linearity in the range of 0 50 microg/L. The characteristic mass and detection limit (3 sigma) were 2.48 and 1.86 pg, respectively. Its relative standard deviation was 2.06% (n = 11). The results of analysis of the National Bureau of standards of America, Standard Reference Material (SRM) No. 1648, Urban Particulate Matter, gave a recovery of 90.15-99.29% with RSD of 4.09%. The method proved to be accurate, sensitive, rapid and simple. PMID- 15825340 TI - [Determination of total amount of acid in soda water by AAS]. AB - A new indirect AAS method for the determination of the total amount of acid in soda water is investigated. After adding MgNH4PO4 reagent to sample solution to react with organic acid, the unreacted reagent subsides by using centrifuge and Mg is detected in solution by AAS. The content of acid is calculated in samples. The results agreed weel with those obtained by volumetric analysis. The relative standard deviations are less than 5.3% and the recoveries are between 91 and 104%. PMID- 15825341 TI - [Study on the atomic fluorescence spectrometric determination of trace bismuth in geological samples by hydride generation in slurry]. AB - A method for atomic fluorescence spectrometric determination of trace bismuth in geological samples by hydride generation in slurry was proposed. The effect of various factors, such as acid media, the particle size of sample and the concentration of slurry etc., on hydride generation was investigated. This method is simple and rapid and has been applied to the determination of trace bismuth in GSD standard samples with satisfaction. PMID- 15825342 TI - [Study on the determination of trace mercury in foods by circulating enrichment cold vapour atomic absorption spectrometry]. AB - The determination of trace mercury using the self-developed circulating enrichment apparatus with cold vapour atomic absorption spectrometry was investigated. Various experimental conditions were discussed and optimized. The effect of interferences was examined. The developed method is sensitive, accurate, simple and available and has already been used in the analysis of various kinds of food samples and national standard reference materials, and the results obtained were satisfactory. PMID- 15825343 TI - [Determination of trace Fe in doxycycline hydrochloric acid by flame atomic absorption spectrophotometry]. AB - This paper reports the development of the determination of trace metal Fe in doxycycline hydrochloric acid by flame atomic absorption spectrophotometry. The method is convenient to determine elements in doxycycline hydrochloric acid. The results show that the method offers good precision and accuracy. The relative standard deviation for Fe was 0.54% and the rate of recovery was 99.2%. PMID- 15825344 TI - [Synthesis and molecular recognition study of 6-Cs-beta-CD]. AB - The reagents 6-Cs-beta-CD with molecular recognition function were synthesized and confirmed. Both basic condition and enough illumination were necessary for obtaining stable and strong fluorescence. With 6-Cs-beta-CD as the host, cyclohexanol, cyclohexene, cyclohexane and bromocyclohexane as the guest, the recognition properties were studied. The affinity of 6-Cs-beta-CD and the guest is distinct with different hydrophobicity of the guest. PMID- 15825345 TI - [A kinetic study of organic reaction by Kalman filter and spectrophotometry]. AB - The Kalman filter-spectrometry was used to the study on the chemical dynamics of alpha-Oxo ketene cyclic dithioacetals. Results show that the method can effectively eliminate or reduce the intercomponent interferences in the simultaneous determination of all components. The method was successfully used to study the condensation reaction dynamics of 1-phenyl-2-(1,2 dithioethylene)methylene-1,3-dibutandione and m-nitrophenyl aldehyde. PMID- 15825346 TI - [Simultaneous determination of zinc, cadmium and lead chelated by porphyrin with partial least squares fluorimetry]. AB - The fluorescence spectra of super-highly sensitive chelate systems containing multicomponent metal porphyrins are analysed by partial least squares method, and a computerized analysis for the simultaneous determination of zinc, cadmium and lead is developed. Zinc, cadmium and lead can quantitatively combine with meso tetrakis-(4-methoxyphenyl)-porphyrin in a strong base medium and in the presence of Tween-80 when they are heated for 30 min in a boiling water-bath. The proposed method has been applied to the simultane ous determination of zinc, cadmium and lead in artificial and environmental water samples, and the recovery ratios are in the range of 93.2-107%. PMID- 15825347 TI - [A fluorescence photometry study on the suberinite in the shallow burial peat]. AB - A detailed study of the suberinite in the shallow burial peat from Zhenan basin, western Yunnan, has been carried out. The study has revealed that the suberinite is the most unstable maceral among all the stable macerals. It may be oxidized and/or decomposed, resulting in changes in the structure and, especially optical feature. The fluorescence light of the suberinite that has not been oxidized and decomposed is green with I546 ranging from 1.5 to 2.0; and the fluorescence spectrum has one peak: lambda(max) ranging from 480 to 510nm; Q500/650 ranging from 0.12 to 0.26; semi-wave width averaging 150nm. Nevertheless, those suberinites that are oxidized and/or decomposed give off yellow or yellowish brown fluorescent light and the I546 reduces to less than 1, even to 0.05 lambda(max) increases from 580 to 610nm; Q500/560 from 2.75 to 3.31. With illuminating time going by, the fluorescence feature of the suberinite could be changed, mainly in the fluorescence intensity and fluorescence spectrum. The suberinite that has been not oxidized and/or decomposed changes negetively in fluorescence intensity and its fluorescence spectrum moves towards red light wavelength. Nevertheless those suberinites that have been oxidized and/or decomposed change positively in fluorescence intensity and the fluorescent spectrum moves towards blue light wavelength. PMID- 15825348 TI - [Determination of ascorbic acid and its stability of Fructus Hippophae fruit by the method of high-valence iron reduction]. AB - The high-valence iron reduction method was used to determine the ascorbic acid of fructus hippophae fruit. The reduction method had simple, high sensitivity and accuracy in comparison with fluorospectrophotometry or indophenol titration method. Its limit of detection was 0.01 microg/mL, the relative standard deviation and the average recovery was 0.83% (n = 8) and 95.6% (n = 6) respectively. We also researched the influence of the environment factors (temperature, storage period and condition, pH value) on the stability of ascorbic acid in fructus hippophae fruit. The ascorbic acid in fructus hippophae fruit was more stable than thepure ascorbic acid on the same condition of heating (80 C,1 hour), in pH neutral or storing in the refrigerator. PMID- 15825349 TI - [Determination of the content of serum calcium with methylthymol blue as chromogenic reagent]. AB - It is reported in this paper that calcium can be determined with MTB as chromogenic reagent by isoabsorption dual-wavelength elimination. The measuring wavelength and referential wavelength were 613.7 and 590.0nm, respectively. No masking reagent was necessary in the analysis. The average recovery and RSD of calcium are 98.98% and 0.81%, respectively. Compared with titration, this method is rapid, simple and precise. PMID- 15825350 TI - [Simultaneous determination of metronidazolum and vitamin B6 by the derivative spectrophotometry]. AB - A new method for simultaneous determination of metronidazolum and vitamin B6 was established by the third derivative spectrophotometry in 0.1 mol/L HCl solution. The detection limits were 0.81 microg/mL for metronidazolum and 0.16 microg/mL for vitamin B6. The linear range of determination was 1.6-22.4 microg/mL for both metronidazolum and vitamin B6. This method has been applied to the determination of metronidazolum and vitamin B6 in tabellae metronidazoli compositae with satisfactory results. The coefficients of variation are below 2.26 and 2.65% and the recoveries are 93-102% and 99-101% for metronidazolum and vitamin 6, respectively. PMID- 15825351 TI - [Flotation spectrophotometric determination of platinum on isochromatic dye ion pair with tetrabromofluorescein and rhodamine 6G]. AB - The flotation spectrophotometric method for the determination of platinum with isochromatic dye ion-pair is described. Rhodamine 6G (R6G) was produced and entered into aqueous phase when pH = 5.5 buffer solution was added to the Pt (SnCl3)5 (R6G)3 by isopropyl ether flotation. And then R6G. TBF was floated by tetrabromofluorescein(TBF)with toluene. The floatation was dissolved in acetone and absorbance of the solution was measured at 530nm. The sensitivity was raised because of the six dyes (Pt : R6G : TBF = 1 : 3 : 3) assistant effect. The molar absorptivity of this method is 5.92 x 10(5)L x mol(-1) x cm(-1). Beer's law was obeyed in the range of 0-2 microg/10mL with a relative standard deviation of 3.8%. The method was applied satisfactorily to the determination of platinum contained in catalysts without separation. PMID- 15825352 TI - [Application of charge transfer reaction in the determination of aminosalicylate sodium]. AB - A spectrophotometric method for the determination of aminosalicylate sodium has been developed based on the charge transfer reaction. The Beer's law is obeyed in the range of 40-160 microg/mL of aminosalicylate sodium. The apparent molar absorptivity of the complex at 660nm is 1.17 X 10(3)L x mol(-1). The average standard deviation of ten determination is 0.67%. The composition of aminosalicylate sodium 2, 6-Dichloroquinone chlorimide complex is 1:1. The proposed method has been applied to the determination of aminosalicylate sodium tablets with satisfactory results. PMID- 15825353 TI - [Application of three-dimension luminescence method to the identification of petroleum pollutants]. AB - As three-dimension luminescence method can provide abundant sample information, it is applied to the identification of petroleum pollutants. In this paper, the whole analytical process of three-dimension luminescence method is described, including sample disposal, analytical conditions, analytical process, especially the automation in acquisition, disposal and differentiation of analytical data. PMID- 15825354 TI - [Determination of Y, Ce, Ba and Cu in Y(1-x) Ce(x),Ba(2)Cu(3)O(y), using x-ray fluorescence fundamental parameter method]. AB - The determination of Y, Ce, Ba and Cu in a series of Y(1-x)Ce(x)Ba(2)Cu(3)O(y) samples has been performed using X-ray fluorescence fundamental parameter method. The composition is given in atomic percentage. The results agree well with those obtained by ICP. It is shown that this method is appropriate for the determination of compositions of the high temperature superconductors. In addition, the influence of oxygen on analytical elements has been discussed in this paper. The fraction of the matrix total mass absorption that is associated with oxygen for each analytical line can not be larger than 3.44%. The determined composition of the samples show different from their nominal composition,which may affect the properties of high temperature superconductor. PMID- 15825355 TI - [Study on a new method for spectrophotometry]. AB - In this paper, the relation between absorbance difference and concentration was measured by the ultraviolet spectrophotometer and computer made P-E company of U.S.A. The basic goal is to prove that the standard absorption curve method can be replaced by the new method, in which the sample was put in standard room and the standard material in sample room. Utilizing this method can minimize the error, most importantly, can measure the higher concentration sample, which can not be measured by the standard curve method. The new method does not require a Linear deltaA-c linear relationship. PMID- 15825356 TI - [Analysis of trace Pd in marine deposit samples using laser-excited atomic fluorescence spectrometry]. AB - Laser-excited atomic fluorescence spectrometry and its experimental set-up are introduced in this paper. Using this spectrometer, Pd contents in 36 marine deposit samples of the East-sea continental shelf were analyzed. The results show that the detection limit and the precision of this spectrometer are 50 pg/mL and 6% respectively, and the measurements of Pd in marine deposit samples are reliable. PMID- 15825357 TI - [A study on the spectra of L-fucose]. AB - The concentration of L-fucose will extraordinarily increase in urine in patients with early stage of cancer. It seems that this is a significant clue for early phase diagnosis of cancer. Unfortunately the sensitive detection of L-fucose is difficult. Thus we carried out researches on IR-spectrum, Raman spectrum, ultraviolet absorption spectrum and ultraviolet fluorescence spectrum. We compared the results from patients and healthy subjects. It is confirmed that the spectroscopic method, especially ultraviolet fluorescence spectrophotometry, may be suitable for early stage screening of some kinds of cancer. This test is rapid, inexpensive and painless. PMID- 15825358 TI - [Photoacoustic spectra of La (III), Gd (III), Tb (III) 8-hydroxyquinoline complexes and relaxation process]. AB - 8-hydroxyquinoline complexes of La(III) Gd (III) and Tb (III) were synthesized and the photoacoustic (PA)spectra of these complexes were measured in the region of 300-800nm at room temperature. The PA spectra and excitation spectra are compared and interpreted. It is observed that the photoacoustic spectra and the excitation spectra behave complementarily. The different fluorescence properties of the complexes and the relaxation process models are studied by the photoacoustic amplitude and phase spectra. It is confirmed by the PA phase spectrum that the PA absorption of pi-pi* transition of Tb (Ox)3 complex is dominantly contributed by the slow radiationless relaxation which is due to inner molecular energy transfer. PMID- 15825359 TI - [Low temperature FTIR study of the structure of ruthenium polyhydrides]. AB - A novel compound of Ru2H4 (CO) (PPh3)4 (A) is synthesized under hydrogen atmosphere. An in situ infrared apparatus which can be operated at liquid nitrogen temperarure is established. The structural conversion of RuH4 (PPh3)3 between dihydrogen and dihydride is observed firstly at room temperature. It is suggested that a fluxional molecule between terminal and bridged hydrogen appears in (A) with temperature changes. H2 molecule coordinated in ruthenium complex is rather easy to be substituted by CO, N2 and small organic molecule, this property is very important for activing molecular hydrogen and using in hydrogenation of unsaturated compounds. PMID- 15825360 TI - [FTIR study on the normal and tumor gastrointestinal tissues]. AB - Series of cancer tissues and corresponding normal tissues of gastrointestinal tract (stomach, colon, esophagus) were studied by FTIR technique and the results showed the analogy of the spectra for the cancer tissues, while the spectra of normal tissues can be classified into three kinds. The secondary structures of protein were obtained by using deconvolution and curve-fitting techniques with Amide I bands and the results showed that the contents of alpha -helix and beta sheet are different between the normal and cancer tissues. PMID- 15825361 TI - [The high resolution spectroscopy of CH35 Cl3 and CH35Cl2(37) V = 3 stretching overtone]. AB - The Fourier transform spectrometer (FTS) is used to record the high resolution spectrum of V = 3 stretching overtone for two isotopic species: CH35Cl3 and CH35Cl2(37)Cl. The rotational constants of CH35Cl2(37) Cl are calculated approximately from its isotope, CH35Cl3. The ground states combination difference method is used to assign the observed transitions. Because of the relatively small rotational constant, the resolution of 0.02 cm(-1) is still not enough to resolve the rotational K structure of CHCl3. So, all the observed transitions are assigned as parallel band transitions arising from the k = 0 rotational states. A weighted non-linear least-square fitting program is developed to fit the assigned transitions and derive the rotational constants. The result of the fitting for the CH35Cl2(37)Cl spectrum indicates that it is reasonable and reliable to derive the spectroscopic parameters using the isotopic effect. PMID- 15825362 TI - [The investigation of interaction for silver(I) ion with gamma-globulin by resonance spectroscopy]. AB - The interaction for silver (I) ion with gamma-globulin has been investigated by resonance spectroscopy. It is shown that 2 or 4 coordination species for amino acid residues of gamma-globulin with Ag (I) ion were formed and their resonance spectra appear to be different in amide bands region. It has been studied that effect of reaction time on resonance Raman(RR) spectra in heat condition. Results showed that the intensity of RR has maximum at about 15 min of reaction, and after the time, Ag (I) ion on species will reduced to elemental silver. PMID- 15825363 TI - [Synthesis of new pyrazoline fluorescent compounds and their IR spectra and fluorescence property]. AB - The Fluorescent compounds of pyrazoline have been used in many field. According to Schellhammer theory, we designed and synthesized five new benzothiazoyl pyrazoline compounds. Their infrared spectra and fluorescent spectra of five compounds were detected. The results showed that five compounds have fluorescence, fluorescence emission wavelength is about 450nm. Effect of different substituents on the fluorescent intensity was investigated. PMID- 15825364 TI - [Spectroscopic investigations on sol-gel transition of alginate solutions by addition of cupric ion]. AB - Viscosity measurements and 13C-NMR, UV/VIS, and Raman spectroscopic studies have been carried out of various concentration of alginate solutions undergoing sol gel transition induced by addition of cupric cation. From analyses of the spectra, a qualitative explaination was suggested of the interaction mode of the coordination between functional groups and cupric ions, the conformational change of polychains and the behaviour of solvent molecules druing the process of sol gel transition. The mechanisms of phase transition of our system induced by the heavy metal ion have been discussed. PMID- 15825365 TI - [Molecular spectra study of fabrication reaction of lead styphnate]. AB - The entire reaction process in the fabrication of lead styphnate in situ was analyzed by using laser Raman spectrometer and FTIR spectrometer, including reaction time determination, testing of the reactants proportions, and identification of the resultants in terms of molecular structure. PMID- 15825366 TI - [Characteristics of IR spectra for oxadiazole]. AB - Infrared spectra of 2,5-o-dimethylbenzene-1,3,4-oxdiazole and 2,5-m dimethylbenzene-1,3,4-oxdiazole were studied. Their principal vibration bands were assigned and the characteristics of their IR spectra were compared with each other. PMID- 15825367 TI - [Study of spectral interferences of REEs with the matrix of dy for a high resolution ICP-AES]. AB - The spectral interferences of the matrix of Dy on other REEs have been studied for a high resolution ICP-AES with a grating of 3600 grooves/mm. 66 analytical lines of 14 REEs have been selected as the prominent lines in the spectral interference studies. Three kinds of databases for Dy lines have been established for the spectra search and recognition. Thus, these databases can provide information of Dy lines accurately and conveniently. PMID- 15825368 TI - [Study of the excitation and diffusion processes of sample atoms in a microwave boosted microsecond-pulse glow discharge source with optical emission spectrometry]. AB - The excitation and diffusion processes of brass sample atoms in a GD-MIP tandem source have been studied with a home-made MIP boosted micros-pulse GD device. Experimental results show that under relatively low discharge pressure (< 180Pa), the micros-pulse GD can couple quite well with MIP and emit strong radiation of analytical lines. When the GD source is operated under a pressure higher than 200Pa, two emission peaks appear independently in time for a given resonant atomic line, because sample atoms are structurally excited separately first by the micros-pulse GD and then by the MIP. According to the two emission peaks, the diffusing velocities of copper atoms and zinc atoms can be calculated, yielding values of 150 and 129m/s, respectively, and the most excited area in-the micros pulse GD is about 1.94-2.25mm away from the sputtering surface of the sample cathode. The effects of discharge parameters on emission intensities have been also investigated. PMID- 15825369 TI - [Determination of trace elements in shark cartilage by inductively coupled plasma atomic emission spectrometry]. AB - Semiquantitative estimation of all elements in shark cartilage was investigated by inductively coupled plasma mass spectrometry (ICP-MS). The determination of trace elements, namely Fe, Zn, Se, Cu, Mn, Mo, Ti and Sr in shark cartilage, was carried out using inductively coupled plasma atomic emission spectrometry (ICP AES). The matrix effects were overcome by using yttrium as an internal standard element. The recoveries are in the range of 81.6 to 100.7%. The determination limits of Fe, Zn, Se, Cu, Mn, Mo, Ti and Sr are 0.60, 0.55, 0.21, 0.39, 0.042, 0.27, 0.038 and 0.48 microg x g(-1), respectively. The results showed that the shark cartilage contains higher amount of Fe, Zn, Se, Cu, Mn, Mo, Ti and Sr than those in other fishes and in other animal bones. PMID- 15825370 TI - [Direct determination of trace elements in tungsten products with ICP-AES]. AB - A direct determination of Al, As, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Sn, Ti and V in tungsten products with ICP-AES is presented. The influence of tungsten matrix is studied. Based on the selected optimum operating parameters, the feasibility of the proposed method is evaluated by analyzing three WO3 National References. The results are satisfactory. PMID- 15825371 TI - [An ICP-AES method for analysis of impurities in zinc and its compounds]. AB - An ICP-AES method for determining Ag, Co, Al, Pb etc. is described. The procedure involves direct determination of zinc and separating zinc from the impurities using coprecipitation with Y (OH)3. The RSD is 1.6-8.1%. PMID- 15825373 TI - [Recent development of studies on interferences and their correction methods in ICP-AES(II)]. AB - In this paper, the spectral interferences in ICP-AES were reviewed continuously. PMID- 15825374 TI - [Determination of sulphate ion in environmental water samples by flow injection spectrophotometry]. AB - Sulphate ion in environmental water samples was successfully determined by flow injection spectrophotometry, based on the reaction with barium dimethylsulphonazo (III) complex. The linear range is 1-4 mg/L, the recoveries are 95-106%, relative standard deviation is 1.5% (n = 10) and the precision of the procedure for long time (about 4h) < 5%, respectively. PMID- 15825372 TI - [Determination of 15 trace rare earth elements in rice by microwave digestion ICP MS]. AB - An ICP-MS method for direct determination of 15 trace rare earth elements in rice was developed. Rapid and complete dissolution of rice was achieved by a microwave digestion procedure with concentrated nitric acid and hydrogen peroxide. The detection limits are 7.0-25.8 pg/mL, the recoveries for spiked samples are 95 105%, RSDs are 2.3-4.2%. This method is rapid, sensitive and accurate. PMID- 15825375 TI - [Extractive chromatograph on-line preconcentration flow injection flame atomic absorption spectrometric determination of trace silver in geological samples]. AB - Extractive chromatograph using a TPP column was applied to preconcentration and matrix separation for trace silver in geological samples. A flow injection on line method for the determination of silver by flame AAS is described. An enrichment factor of 20 was achieved at a sampling frequeucy of 30 times/h. The precision RSD is 2.0% for the analysis of a geochemical standard reference material (GSD-12, n = 11) and the detection limit obtained (3sigma) is 0.02 microg/g. PMID- 15825376 TI - [Determination of trace lead in copper by enhancing effect FAAS-using derivative technique]. AB - A new method for the determination of trace lead in copper is proposed by enhancing effect-flame atomic absorption spectrometry using derivative technique. Enhancing effect of matrix Cu on lead was investigated. The sensitivity for lead was enhanced 17 times than that of conventional flame atomic absorption spectrometry by combining derivative technique with matrix Cu enhancing effect. Determination result is satisfactory by this method with 0.007 microg/mL detection limit and 0.7% RSD. PMID- 15825378 TI - [Determination of Sb in the resin and products of PET by graphite furnace atomic absorption spectrometry]. AB - The method of determing Sb in resin and products of PET by graphite furnace atomic absorption spectrometry was studied. The sample was pretreated by 4% acetic acid and determined by graphite furnace atomic absorption spectrometry. The sensitivity of this method was 30.5 pg/1% absorption, the relative standard deviation was 3.12% and the rate of recovery was 91.68-97.74%. The results obtained by this method agreed with those of malachite green spectrophotometry. PMID- 15825377 TI - [AAS determination of trace lead, cadmium, copper and nickel by enrichment with 2 mercaptobenzimidazole chelating cellulose]. AB - Chelating cellulose is of simple preparation,excellent nature and low cost. MBM chelating cellulose was prepared with good chelating reagent (2 mercaptobenzimidazole) grafted on cotton by indirect etherification method. Four heavy metal ions in one solution can be simultaneously preconcentrated and eluted under good condition. Results have shown that the approach for separating and preconcentrating trace of Cu, Pb, Cd and Ni has advantages of high exchanging speed, high preconcentration multiple, high selectivity, easy elution and excellent results. PMID- 15825379 TI - [Grazing techniques and x-ray fluorescence analysis]. AB - X-ray analysis has become a new analysis method of material and thin film. Using this method, we can obtain much information on material such as density, layer thickness, interface roughness and compositional depth profile. Application and development of grazing technique in X-ray fluorescence analysis are introduced in this paper. PMID- 15825380 TI - [Study on the spectra of granite of asbestos red and sesame white]. AB - In this paper, the composition and spectra of granite of asbestos red and sesame white are studied by ICP-AES, XRF, IR and X-ray diffraction. According to the X ray diffraction, the components were determined by computer. Comparing results, it is concluded that the red colour of asbestos red granite is caused by the red minerals of Fe contained in it. PMID- 15825381 TI - [Phenomena of supramolecular assembly fluid room temperature phosphorescence (II) -effects of alcohols on room temperature phosphorescence of cyclodextrin/4-iodo 4'-ethyl-biphenyl/bromocyclohexane system]. AB - Beta-cyclodextrin( beta/-CD)can induce 4-iodo-4'-ethyl-biphenyl(IEBP)to emit room temperature phosphorescence(RTP) in the presence of bromocyclohexane (BCH). If butyl alcohol (BuOH) is presented as a fourth component at the same time, RTP intensity of the system is enhanced obviously. Effects of different alcohols, CDs and derivatives of beta-CD on RTP emission of the system are studied. The results show that a ternary host-guest inclusion complex is formed among beta-CD, IEBP and BCH, but the alcohol hydroxyl bonds to the rim of the CD cavities by hydrogen bonding and the alkyl end of alcohol flips over the two tops of the CD due to hydrophobic interaction force. Accordingly, the alcohol acts as a lid for CD cup, thereby shielding IEBP from oxygen and decreasing the collision quenching of oxygen, and decreasing the micro polarity of the CD cavity and the formation constant of ternary inclusion complex increases. As a result, the RTP of the system is enhanced. As far as we can see that the system studied is a typical supramolecular assembly one and the BuOH acts as a molecular regulation for RTP emitting. PMID- 15825382 TI - [Analysis of alpha-Oxo ketene cyclic dithioacetals compounds by using Kalman filtering and UV spectrophotometry]. AB - The Kalman filtering and UV spectrophotometry have been used to analyte the alpha Oxo ketene cyclic dithioacetal compounds in the present paper. The innovation sequence and concentration trends in the Kalman filtering process have been investigated, results show that the combination of the Kalman filtering and UV spectrophotometry is an applicable and reliable means in analysis of this kind of compounds. PMID- 15825383 TI - [A kind of method for measuring absorbance spectrum with dual beam light and one detector]. AB - A kind of method of measuring spectral transmittivity with source compensated is introduced in this paper. It detects simultaneously intensity of source and transmission light with dual light road and one detector by means of the characters of selective frequence amplifing and coherent detection of a lock-in amplifier. The experimental result shows that the method is excellent on overcoming shake of a source and improving signal-to-noise ratio. PMID- 15825384 TI - [Study on the interaction between Cu2+ and cholesterol by spectrometric analysis]. AB - With four kinds of spectrometric analysis methods, i.e. spectrophotometric analysis, AAS, ICP-AES and IR, the interaction between Cu2+ and cholesterol in aqueous solution has been examined. The same results from the different methods showed that there was a surface adsorption between Cu2+ and cholesterol and this interaction would be prominent in physiology-like situation. PMID- 15825385 TI - [Simultaneous determination of phenol and resorcinol by bouble wavelength standard additions method]. AB - In this paper, double wavelength standard additions method was first used in fluorescence analysis. The simultaneous determination of phenol and resorcinol could be carried out when the ratio of phenol to resorcinol concentrations was in the range of 15:1-1:15. The method gives satisfactory results. PMID- 15825386 TI - [Determination of paeoniflorin in Huoxuezhuyu powder]. AB - Thin-layer chromatogram was used to separate paeoniflorin from other compositions in Huoxuezhuyu powder. The content was determined by UV spectrophotometry. The mean recovery was 98.74%, and CV was 1.43%. This method could be used for other compound preparations of traditional Chinese medicines. PMID- 15825387 TI - Proposed standardized method for testing and interpreting susceptibility of Bacteroides fragilis to tetracycline. AB - One hundred twenty-four strains of Bacteroides fragilis were examined for susceptibility to tetracycline disks and by minimal inhibitory concentration (MIC) determinations. MIC values and zone sizes around 30-mug tetracycline disks were determined by using selected test conditions which included Mueller-Hinton agar supplemented with sheep blood, vitamin K, and hemin and an incubation temperature of 35 C in an atmosphere of 80% N(2), 10% H(2), and 10% CO(2). Strains were separated into two distinct populations by geometric mean MIC and dis tests. Of the 124 strains, 78 were resistant and 46 were susceptible. The resistant strains had geometric mean MIC's of 8 mug/ml or greater, whereas the geometric mean MIC's of sensitive strains were 5 mug/ml or less. The disk test proved to be more reproducible than the MIC test and completely separated the resistant and susceptible populations. An interpretive scheme for B. fragilis to tetracycline was statistically derived on the basis of the distribution of zone sizes of susceptible and resistant strains: resistant, 18 mm or less; indeterminant, 19 to 20 mm; and susceptible, 21 mm or greater. These zone sizes compared closely with the Kirby-Bauer criteria for aerobic bacteria. PMID- 15825388 TI - Screening method of agents against the R factor by the use of an Hfr made by integrative suppression with an R factor. AB - By the use of an Hfr strain made by integrative suppression of a derepressed R factor, R100-1 in a dnaA-ts mutant of Escherichia coli and several other appropriate control strains with and without the R factor, a screening system was established which can be used to detect agents which are either inhibitory to the replication of the R factor and lead to its "curing" or are preferentially inhibitory to the growth of the R-carrying bacteria. The validity of this system was shown with several agents already known to be experimentally effective. PMID- 15825389 TI - Human pharmacology of 6-[D-alpha-(3-guanylureido)-phenylacetamido]-penicillanic acid (BL-P1654). AB - BL-P1654 is a new alpha-substituted ureido penicillin which has broad-spectrum activity. Doses of 0.5 and 1.0 g administered by rapid intravenous injection produce mean peak serum concentrations of 40.7 and 80.8 mug/ml, respectively. The same doses administered by 1-h intravenous infusions produce mean peak serum concentrations of 22.6 and 47 mug/ml, respectively. The mean serum concentrations from 1 to 6 h after onset of administration of the drug are the same for both schedules. The mean serum concentration at 6 h after the rapid intravenous injection of 0.5- and 1.0-g doses of BL-P1654 are 3.4 and 5.4 mug/ml, respectively. A dose of 1.0 g every 4 h maintains a serum concentration of about 15 mug/ml. About 50 to 65% of the BL-P1654 is excreted in the urine during the first 6 h. PMID- 15825390 TI - Rifampin: inhibition of ribonucleic acid synthesis after potentiation by amphotericin B in Saccharomyces cerevisiae. AB - The inhibition of Saccharomyces cerevisiae growth by amphotericin B and rifampin was studied. Rifampin alone had no effect on growth or macromolecular syntheses. Lethal amounts of amphotericin B produced a late inhibition of ribonucleic acid (RNA) synthesis simultaneous with the arrest of growth and protein synthesis. In contrast, low doses of amphotericin B along with rifampin caused an early arrest of RNA synthesis, followed by a later arrest of growth and protein synthesis. Used with rifampin, amphotericin B thus appears to increase cell permeability for rifampin, which in turn inhibits RNA synthesis; such results are consistent with some reports of inhibition of yeast RNA polymerase function by rifampin. Experiments with petite mutants ruled out any special effect of the antibiotics on mitochondrial RNA synthesis, so that nuclear RNA synthesis is affected. Acrylamide gel analyses of RNA pulse-labeled after addition of the two antibiotics in synergy showed that synthesis of all major classes of RNA was progressively and uniformly inhibited. PMID- 15825391 TI - Molecular basis for the selective toxicity of amphotericin B for yeast and filipin for animal cells. AB - Among the polyene antibiotics, many, like filipin, cannot be used clinically because they are toxic; amphotericin B, however, is useful in therapy of human fungal infections because it is less toxic. Both the toxicity of filipin and the therapeutic value of amphotericin B can be rationalized at the cellular and molecular level by the following observations: (i) these polyene antibiotics showed differential effects on cells; filipin was more potent in lysing human red blood cells, whereas amphotericin B was more potent in inhibiting yeast cell growth; and (ii) the effects of filipin were more efficiently inhibited by added cholesterol, the major membrane sterol in human cells, whereas the effects of amphotericin B were more efficiently inhibited by ergosterol, the major membrane sterol in yeast. The simplest inference is that the toxicity and effectiveness of polyenes are determined by their relative avidities for the predominant sterol in cell membranes. PMID- 15825392 TI - Immunological responsiveness of tuberculosis patients receiving rifampin. AB - Rifampin has been shown to impair both humoral and cell-mediated immune responses in animal models. In order to detect a similar effect in man, 11 patients with active tuberculosis were evaluated before, 2 weeks after, and 12 to 16 weeks after initiating rifampin. Several parameters were serially measured including blood lymphocytes, intradermal response to intermediate strength tuberculin (PPD), and in vitro proliferative responses to phytohemagglutinin (PHA), a nonspecific mitogen, and to the specific antigens, PPD and influenza A. No changes in lymphocyte counts were noted. No changes in response were noted 2 weeks after beginning treatment with rifampin. However, compared with initial and 2-week responses, the PHA response was reduced by 47%, the PPD by 68%, and the influenza by 75%, and 6 of 11 patients showed no induration after tuberculin skin testing at the 12- to 16-week point. These results indicate that in doses employed for the treatment of tuberculosis, rifampin has an immunosuppressive effect in man that develops gradually. PMID- 15825393 TI - Alterations in dihydropteroate synthetase in cell-free extracts of sulfanilamide resistant Neisseria meningitidis and Neisseria gonorrhoeae. AB - Extracts from Neisseria meningitidis and N. gonorrhoeae with varying susceptibility to sulfanilamide have been investigated for dihydropteroate synthetase activity. Sulfanilamide was a competitive inhibitor of dihydropteroate synthetase with respect to p-aminobenzoate in extracts from both species. Though the K(m) for p-aminobenzoate was unaffected, the K(i) for sulfanilamide increased and the V(max) decreased as the strains' resistance to sulfanilamide increased. Temperature studies have revealed differences in the dihydropteroate synthetase from N. meningitidis and N. gonorrhoeae. A direct relationship was observed between the minimal inhibitory concentration of sulfanilamide determined in vitro and the ratio of K(i)/K(m). This ratio may be a molecular explanation of sulfanilamide resistance for both N. meningitidis and N. gonorrhoeae. PMID- 15825394 TI - Effects of some antiviral isatinisothiosemicarbazones on cellular and viral ribonucleic acid synthesis in Mengovirus-infected FL cells. AB - Three antiviral isatinisothiosemicarbazones strongly inhibited the incorporation of [(3)H]uridine into the ribonucleic acid (RNA) of FL cells as a consequence of the inhibition of uridine transport. After prelabeling of cells at a low temperature (1 h at 16 C) with uptake of [(3)H]uridine into the acid-soluble nucleotide pool, the later addition of the test compounds revealed only a small or negligible influence on host-directed RNA synthesis. The pulse-labeled soluble nucleotide pool of FL cells was sufficient to give a gradual increase in incorporation into RNA over a period of 7 h. With the same method of prelabeling at the beginning of the experiment, it was also possible to detect virus-induced RNA synthesis in the presence of actinomycin D. In this way the specific inhibitory action of the three isatinisothiosemicarbazones on viral RNA synthesis could be demonstrated. PMID- 15825395 TI - Inactivation of herpes simplex virus by thiosemicarbazones and certain cations. AB - We demonstrated that herpes simplex virus types 1 and 2, including a type 2 strain which transforms hamster cells in vitro, and Herpesvirus saimiri are inactivated by exposure to thiosemicarbazones. Because thiosemicarbazones are thought to interact with heavy metals in this inactivation process (9), we tested and found some of these herpesviruses to be susceptible to exposure to certain heavy metals. A virion polymerase was sought because the ribonucleic acid (RNA) dependent deoxyribonucleic acid (DNA) polymerase of Rous sarcoma virus and the DNA-dependent RNA polymerase of vaccinia virus are inhibited. However, neither DNA nor RNA polymerase activity could be demonstrated in herpes simplex virions. The ability of thiosemicarbazone to ameliorate the course of herpes simplex virus infection in rabbit eyes was observed, but was considered insufficient to be of clinical importance. PMID- 15825396 TI - Evaluation of isoprinosine in experimental human rhinovirus infection. AB - The prophylactic efficacy of isoprinosine was evaluated in a double-blind fashion in volunteers challenged with two types of rhinovirus. In the rhinovirus 44 and 32 trials, each of 9 men received a placebo, and eight and 11 men received the drug, respectively. Oral isoprinosine, 6 g a day, was given for 2 days prior to intranasal challenge with 100 mean tissue culture infective doses of the virus and for 7 postchallenge days. In both trials the occurrence and severity of colds were greater in the placebo group, but the difference between the two groups was not significant. Higher antibody titers for both viruses and a greater number of rhinovirus 32 isolations were demonstrated in the drug group but without statistically significant differences. The prophylactic isoprinosine treatment may suppress the cold syndrome, but its effect was not convincingly apparent. PMID- 15825397 TI - Treatment of poxvirus infections in rabbits with 9-beta-D arabinofuranosyladenine. AB - The antiviral efficacy of 9-beta-d-arabinofuranosyladenine (ara-A) was evaluated in localized lesions produced by the intradermal inoculation of rabbits with vaccinia virus (VV) and rabbit Shope fibroma virus (SFV). Ara-A administered intraperitoneally suppressed or significantly reduced the cutaneous pustular lesions produced by VV as well as the benign skin tumors caused by the SFV. With a daily dose of 300 mg/kg given for 5 days starting at the time of infection, or with 600 mg/kg daily starting 3 days after inoculation, we were able to suppress completely the formation of tumors induced by the SFV. The appearance of pustular lesions induced by VV was completely suppressed by a dose of 600 mg of ara-A per kg given for 3 days when the treatment was initiated at the time of infection, but a significant reduction in the number of pustular lesions was obtained with a single dose of 600 mg/kg, or with five doses of 300 mg/kg starting 24 h after inoculation. No toxic effect of ara-A was noted in the treated rabbits. PMID- 15825398 TI - Enhanced toxicity for mice of 6-mercaptopurine with bacterial endotoxin. AB - The toxicity of 6-mercaptopurine was potentiated by 2 mg of either Escherichia coli 026:B6 B endotoxin or Salmonella typhosa 0901 W endotoxin per kg. Nonlethal doses of heat-killed, gram-negative bacteria were also capable of potentiating the lethality of 6-mercaptopurine (6-MP). Salmonella minnesota S, the wild-type strain, and S. minnesota Re 595, a mutant containing only the lipid A and 2-keto 3-deoxyoctonate moiety of the endotoxin molecule, exhibited the same capability to enhance the toxic action of 6-MP. Endotoxin (lipopolysaccharide [LPS]) did not affect the clearance of 6-MP from the circulation, but did alter its apparent metabolism as indicated by blood levels of a metabolite, 6-thiouric acid. The concentration of blood urea nitrogen (BUN) in mice 18 h after injection of 100 mg of 6-MP per kg simultaneously with 2 mg of LPS per kg was significantly elevated over normal values. However, these BUN values were significantly less than those resulting from the administration of one mean lethal dose of either agent. The clearance from the circulation of the gram-negative organism E. coli HB, or the gram-positive organism Staphylococcus epidermidis S, was not affected by 6-MP. Endotoxin had no effect on the clearance of S. epidermidis S, but inhibited that of E. coli HB. When 6-MP and LPS were administered simultaneously with either bacterial species, only the clearance of E. coli HB was inhibited. Mice were protected from the lethality associated with combinations of 6-MP and LPS by (i) prior treatment with phenobarbital, (ii) caffeine, (iii) methylprednisolone, and (iv) polymyxin B sulfate. With the exception of caffeine, each regimen protected mice against the lethal effects of 400 mg of 6-MP per kg, and methylprednisolone or polymyxin B protected mice against 8 mg of LPS per kg. PMID- 15825399 TI - Studies on the mechanism of action of miconazole: effect of miconazole on respiration and cell permeability of Candida albicans. AB - The antifungal drug, miconazole nitrate, inhibits the growth of several species of Candida. Candida albicans, one of the pathogenic species, was totally inhibited at a concentration of approximately 10 mug/ml. Endogenous respiration was unaffected by the drug at a concentration as high as 100 mug/ml, whereas exogenous respiration was markedly sensitive and inhibited to an extent of 85%. The permeability of the cell membrane was changed as evidenced by the leakage of 260-nm absorbing materials, amino acids, proteins, and inorganic cations. The results we present clearly show that the drug alters the cellular permeability, and thus the exogenous respiration becomes sensitive to the drug. PMID- 15825400 TI - Effect of certain polypeptides on the biological activities of Measles virus. AB - The effect of a selected tripeptide, Z-d-Phe-Phe-Arg(NO(2)) (SV-4814) on the kinetics of lysis of green monkey erythrocytes by measles virus hemolysin suggested conditions of competitive inhibition. SV-4814 prevented not only hemoglobin release, but also the preceding leakage of K(+) through the cell membrane. The capacity of 12 different dipeptides, 6 tripeptides, including SV 4814, and 4 tetrapeptides to inhibit measles virus-specific hemolysis and infectivity was analyzed. Tri- and tetrapeptides were more active than dipeptides. The presence of aromatic amino acids and a free carboxyl terminal appeared to be a prerequisite for inhibitory activity. Two compounds, one tripeptide (Z-Phe-d-Phe-d-Tyr) and one tetrapeptide [Z-d-Phe-Arg(NO(2))-d-Met], carried an inhibitory activity, which was about two times higher than that of SV 4814. PMID- 15825401 TI - Regulation of the formation of alkaline phosphatase during neomycin biosynthesis. AB - The use of a washed-cell system to study the factors controlling the synthesis of alkaline phosphatase and neomycin by Streptomyces fradiae has shown that calcium and magnesium salts are stimulatory, with maximal synthesis of both achieved with a combination of these salts. Among all the carbon sources studied, only arabinose induces alkaline phosphatase synthesis, whereas glucose and other carbon sources inhibit the synthesis of the enzyme. Asparagine is a very good inducer of enzyme and neomycin synthesis, with lysine and alanine having lower stimulatory effects. The appearance of alkaline phosphatase is due to de novo protein synthesis as demonstrated by the inhibition of its synthesis in the presence of chloramphenicol. There is a good correlation between the synthesis of alkaline phosphatase and neomycin biosynthesis. PMID- 15825402 TI - Demonstration of R factors from Pseudomonas aeruginosa. AB - Sixty strains of Pseudomonas aeruginosa were selected from our clinical stock cultures and used as donors of drug resistance to examine the presence of R factors. They were highly resistant to one or more of six drugs including tetracycline (TC), chloramphenicol (CM), streptomycin (SM), sulfanilamide (SA), carbenicillin (CPC), and kanamycin (KM). Three FP(-), auxotrophic mutants of P. aeruginosa were used as recipients, which had been made resistant to either rifampin (RMP) or KM by in vitro mutation. Each donor was mated with each of the three recipients, and 45 strains were found to transfer their resistance to one or more recipients. A second series of conjugation experiments using 45 exconjugants as donors showed that 23 could transfer their resistance. We conclude that these strains carry R factors that are transmissible to one or more recipients. Eleven carried resistance to (TC.CM.SM.SA.CPC), nine to (TC.CM.SM.SA), one to (TC.CM.SA.CPC), one to (TC.CM.SA), and one to (SM). All of these R factors could not be transmitted to other species belonging to the family Enterobacteriaceae. PMID- 15825403 TI - Inactivation of dihydrostreptomycin and spectinomycin by Staphylococcus aureus. AB - Three types of Staphylococcus aureus strains were isolated with respect to streptomycin (SM) and spectinomycin (SPC) resistance, namely, SM(r)SPC(r), SM(r)SPC(s), and SM(s)SPC(r) (r, resistant; s, sensitive). Curing experiments and transduction analysis of strain MS7990 (SM(r).SPC(r).EM(r)) (EM, erythromycin) disclosed that the loci governing SM and SPC resistance are different and exist on different nontransferable plasmids (r factor), one plasmid carrying the genes governing SM resistance and another possessing the genes governing resistance to both SPC and EM. Strain MS7990 (SM(r).SPC(r)) inactivated both drugs by adenylylation. Similarly, the SM(r)SPC(s) and SM(s)SPC(r) strains inactivated SM and SPC, respectively, by adenylylation, although the adenylylated positions of both drugs have not been established as yet. The adenylylated SM in staphylococci was shown to be different from 3''-adenylyl-SM, indicating the possibility of the existence of a different enzyme from SM3''-adenylyl transferase demonstrated in Escherichia coli strains. PMID- 15825404 TI - Enzymatic inactivation of a new aminoglycoside antibiotic, sisomicin by resistant strains of Pseudomonas aeruginosa. AB - The antibacterial activity of sisomicin (SS), a new aminoglycoside antibiotic active toward clinical isolates of Pseudomonas aeruginosa, was determined and compared with that of the gentamicin C complex. Both drugs were effective against these strains and showed almost the same antibacterial activity. A few strains were found to be resistant to SS. The antibiotic was inactivated by a cell-free extract from the SS-resistant strains due to acetylation of the drug. Comparative studies of the inactivation of the drugs which lack a 6'-amino group in the amino sugar linked to 2-deoxystreptamine strongly suggested that SS inactivation was due to acetylation of the 6'-amino group of the 4',5'-didehydropurpurosamine moiety. PMID- 15825405 TI - Alleged effect of bile constituents on gentamicin assays. AB - It has been claimed that gentamicin assays on the serum of jaundiced patients give falsely low levels and suggested that the antibiotic in such sera is bound to bile acids and thus is partly inactivated. No evidence of such inactivation has been obtained (i) in the serum of jaundiced patients treated with gentamicin, (ii) in sera with high bilirubin contents to which gentamicin was added, or (iii) when bile or bile acids were added with gentamicin to normal serum. PMID- 15825406 TI - Gentamicin-adenylyltransferase activity as a cause of gentamicin resistance in clinical isolates of Pseudomonas aeruginosa. AB - Gentamicin adenylyltransferase activity was found in extracts of clinical isolates of gentamicin-resistant Pseudomonas aeruginosa. Extracts of one of these isolates, P. aeruginosa POW, inactivated gentamicin in the presence of adenosine 5'-triphosphate. Extracts of strain POW catalyzed the binding of radioactivity from [(14)C]adenine adenosine 5'-triphosphate to gentamicin components, tobramycin, sisomicin, kanamycin A and B and, to a variable degree, streptomycin and spectinomycin. The substrate profile with these agents and other aminocyclitols was similar to that obtained with R factor-mediated gentamicin adenylyltransferase found in Enterobacteriaceae. Adenylylating activity was absent in gentamicin-susceptible mutants of strain POW. Adenylylation may be added to acetylation as an enzymatic mechanism responsible for gentamicin resistance among strains of P. aeruginosa. PMID- 15825407 TI - Synergy between cephalosporin and aminoglycoside antibiotics against Providencia and Proteus. AB - Clinical isolates of Providencia and Proteus with relative aminoglycoside resistance were tested for susceptibility to combinations of gentamicin or tobramycin with cephalothin or cefazolin. The minimal bactericidal concentration of aminoglycoside for one-third of the strains was reduced by fourfold or more in the presence of one-fourth of the minimal bactericidal concentration of either cephalosporin. This effect was achieved by clinically attainable concentrations of cephalothin or cefazolin. PMID- 15825408 TI - New antibiotic produced by bacteria, 5-beta-D-xylofuranosylneamine. AB - A new aminoglycoside antibiotic was isolated from the fermentation broths of two strains of Bacillus species. The antibiotic is active against gram-positive and some gram-negative bacteria, and its antimicrobial spectrum is similar to that of ribostamycin. The chemical structure was determined to be 5-beta-d xylofuranosylneamine, which is identical to the deacylated product obtained from butirosin A. PMID- 15825409 TI - Mechanism of silver sulfadiazine action on burn wound infections. AB - The role of silver and sulfadiazine in the mechanism of action of silver sulfadiazine on burn wound infections was investigated. Silver, but not sulfadiazine, was bound by bacteria. Sulfadiazine did not act as an antibacterial agent in low concentrations, but exhibited specific synergism in combination with subinhibitory levels of silver sulfadiazine. The efficacy of silver sulfadiazine is thought to result from its slow and steady reactions with serum and other sodium chloride-containing body fluids, which permits the slow and sustained delivery of silver ions into the wound environs. In this circumstance, a relatively minute amount of sulfadiazine appears active. PMID- 15825410 TI - Susceptibility of Clostridium ramosum to antimicrobial agents. AB - The susceptibility of 49 strains of Clostridium ramosum to 10 antibiotics was determined by agar dilution and disk diffusion tests. Results showed that, among the anaerobes, C. ramosum is second only to Bacteroides fragilis in its resistance to antimicrobial agents. All strains were susceptible to penicillin, carbenicillin, chloramphenicol, vancomycin, and metronidazole at readily achievable blood levels. Most strains (83%) were susceptible to erythromycin. There was a high level of resistance to clindamycin in 16% of the strains. All isolates were resistant to rifampin and gentamicin, and most were resistant to lincomycin. Assessment of susceptibility by measurement of inhibition zone diameters with disk diffusion tests was not satisfactory. PMID- 15825411 TI - Plasmid-mediated production of staphylococcin in bacteriophage type 71 Staphylococcus aureus. AB - Staphylococcin production by Staphylococcus aureus strain C55 was eliminated when producing strains were exposed to various curing treatments. Acridine orange, ethidium bromide, and sodium dodecyl sulfate each effected less than 15% cure rates in treated cells. Rifampin eliminated the staphylococcin in 83% of treated isolates, and growth at 42 C resulted in a 97% cure rate. Curing of staphylococcin production and elimination of penicillinase plasmid were independent of one another. Several characteristics of the cured strains were examined and compared with the bacteriocin-producing strains. Whereas bacteriocin producing cells were resistant to the bactericidal action of the staphylococcin and failed to adsorb it, cured cells adsorbed the staphylococcin and were susceptible to its lethal effect. PMID- 15825412 TI - Comparison and evaluation of carbenicillin disks in diffusion susceptibility testing. AB - A broad variety of bacterial strains, including 79 Pseudomonas aeruginosa, were studied in an in vitro evaluation of carbenicillin disk susceptibility testing. Regression analysis with both 50-mug and 100-mug carbenicillin disks was carried out. Organisms having minimal inhibitory concentration values of 100 to 200 mug/ml demonstrated zones of less than 11 mm with the 50-mug disk, resulting in very little opportunity for appropriate discrimination of results. The line of regression for the 50-mug disk intersected the ordinate at a point just above the minimal inhibitory concentration value considered to be the limit of intermediate susceptibility for Pseudomonas. These considerations, together with evidence of greater disk content variation in the 50-mug than in the 100-mug disks assayed, considerable manufacturer-to-manufacturer variability with the 50-mug disk, and the more appropriate performance of the 100-mug disk, lead us to conclude that the 100-mug disk better serves the clinical test requirements for this agent than does the 50-mug disk, which is currently the only disk available for laboratory testing. PMID- 15825413 TI - Potential unreliability of nitrofurantoin disks in susceptibility testing. AB - The Kirby-Bauer disk diffusion antimicrobial susceptibility test was studied with three lots of commercial nitrofurantoin disks by using 50 strains of Proteus mirabilis. One lot of disks consistently gave zone diameters 3 to 6 mm larger than the other two and resulted in a substantial number of Proteus mirabilis strains being erroneously categorized as sensitive. Antimicrobial content in disks of the three lots did not account for these differences. The pH of eluate from the unreliable disks was higher than from disks of the other two and resulted in more rapid diffusion of the antimicrobial and consequently larger zones of inhibition. PMID- 15825414 TI - Pharmacokinetics of tobramycin in patients with stable renal impairment, patients undergoing peritoneal dialysis, and patients on chronic hemodialysis. AB - The pharmacokinetics of tobramycin were studied in five patients with stable renal impairment, four patients requiring peritoneal dialysis, and four patients on chronic hemodialysis. The half-life of the drug varied with the level of the serum creatinine in the first group of patients, and the average volume of distribution was 15 liters. Only 49% of the administered dose of tobramycin was recovered during 36 h of peritoneal dialysis. The average clearance of tobramycin during hemodialysis was 49.1 ml/min, and 51.5% of the administered dose was recovered during a 6-h dialysis. PMID- 15825415 TI - Chromatography of penicillins, penicilloates, and penicilloylamides on dextran gels. AB - The factors influencing the chromatographic behavior on dextran gels of penicillins and their derivatives were investigated by comparing elution profiles and partition coefficients (K(D) and K(AV)) of penicillins differing in side chain structure and among penicillin derivatives of identical side-chain but different nuclear structure. Under the conditions of pH and ionic strength employed (pH 7.4, 0.145 M NaCl, 0.05 M PO(4)), side-chain adsorptive effects best explained the anomalous behavior of benzylpenicillin and of oxacillin and its chlorine-substituted analogues. Polar side-chain substituents, such as the amino group of ampicillin and the carboxyl group of carbenicillin, and cleavage of the beta-lactam ring, exemplified by penicilloates and penicilloylamines, both appeared to interfere with side-chain-directed adsorption. The differential adsorption of penicillins and their derivatives to dextran gels is not only of theoretical interest relative to the mechanism of chromatography but of practical application to analytical and preparative procedures in penicillin chemistry. PMID- 15825416 TI - Technique for determining the bactericidal effect of drug combinations. AB - Two paper strips, each containing different antimicrobial agents, were placed on plates on Mueller-Hinton agar to permit antibiotic to enter the agar. A filter membrane was placed on this plate, and the microorganisms were planted on the membrane. After 6 h of incubation at 37 C, the membrane was transferred to antibiotic-free Mueller-Hinton agar containing triphenyltetrazolium hydrochloride and incubated for 18 h at 37 C. Specific growth patterns were indicative of additive (indifferent), synergistic, or antagonistic effects of the drug combination used. Trimethoprim and sulfamethoxazole proved to act synergistically against 85% of Escherichia coli, 86% of Klebsiella, and 89% of Proteus mirabilis strains tested. A few strains resistant to either drug were susceptible to their combination. The technique was useful against organisms with widely differing susceptibilities to the two antimicrobial agents tested. PMID- 15825417 TI - Antibiotic produced by Fusarium equiseti NRRL 5537. AB - Fusarium equiseti NRRL 5537 grown on an autoclaved white corn grit medium for 3 to 4 weeks at room temperature produced a substance in excess of 5 g/kg of substrate that inhibited some gram-positive bacteria including mycobacteria. Most Bacillus subtilis, Mycobacterium phlei, and Staphylococcus aureus strains were inhibited when 1 mug of the antibiotic per ml was incorporated into the culture medium. Except for Neisseria perflava, gram-negative bacteria, yeasts, and molds were not inhibited by 128 mug/ml. The antibiotic was recovered as a white powder, had a melting point of 65 to 66 C, and had an intraperitoneal mean lethal dose in white mice of 63 mg/kg of body weight. In thin-layer chromatographic analysis the compound appeared as a single spot in two different solvent systems. Mass spectrometry determined that the molecular weight of the antibiotic was 373 with a molecular formula of C(22)H(31)NO(4). Chemical microanalysis was in accord with the formula. PMID- 15825418 TI - Alteration of contractile function and calcium ion movements in vascular smooth muscle by gentamicin and other aminoglycoside antibiotics. AB - Experiments were conducted to examine the effects of certain aminoglycoside antibiotics on contractile responses and related calcium ion (Ca(2+)) movements in isolated vascular smooth muscle. Gentamicin, kanamycin, and streptomycin decreased contractile responses produced by norepinephrine, histamine, and high K(+) in rabbit aortic strips. The inhibitory action of these antibiotics on mechanical function was more pronounced when the Ca(2+) concentration of the bathing solution was decreased from 1.5 mM (normal Ca(2+) solution) to 0.05 mM (low Ca(2+) solution). The uptake of radiocalcium ((45)Ca) into the isolated media-intimal layer of rabbit aortae was decreased in a maintained manner by each antibiotic. With gentamicin, the inhibitory effect on (45)Ca uptake was shown to be dependent upon the concentration of gentamicin employed and to be more evident in a 0.1 mM Ca(2+) solution than in a normal Ca(2+) solution. In addition, the rate of (45)Ca efflux from the rabbit aortic media-intimal layer was increased in a sustained manner by gentamicin, streptomycin, and kanamycin. Furthermore, contractile responses induced by high K(+) and norepinephrine in canine carotid arterial strips were inhibited by gentamicin. Present findings indicate that aminoglycoside antibiotics interfere with Ca(2+)-linked events leading to activation of the contractile mechanism of vascular smooth muscle. These in vitro findings may partially explain the occurrence of in vivo cardiovascular depression that has occasionally been observed after the administration of chemically related antimicrobial agents. PMID- 15825419 TI - Effect of clindamycin on acute and chronic toxoplasmosis in mice. AB - The effect of clindamycin on survival of mice during acute infection with the RH and C56 strains of Toxoplasma and the ability of this drug to prevent congenital transmission during the acute stage of the infection in the mother and to eradicate the parasite from tissues of mice chronically infected with the C56 strain were evaluated. The drug effectively prevented death due to the acute infection and, in the experimental model employed, eradicated the organism at least from the liver, spleen, and brain of approximately 30 to 50% of the acutely infected animals which survived. Clindamycin also effectively prevented congenital transmission during the acute infection in the mother. During short term treatment (7 days), persistent parasitemia in the chronic infection was effectively diminished or eradicated. More prolonged treatment resulted in a significant clearing of the organisms from the spleens and livers, but not from the brains, of chronically infected mice. PMID- 15825420 TI - In vitro antiviral activity of 6-substituted 9-beta-D-ribofuranosylpurine 3', 5' cyclic phosphates. AB - A series of twelve recently synthesized 6-substituted derivatives of 9-beta-d ribofuranosylpurine 3',5'-cyclic phosphate (RPcMP) were evaluated for in vitro antiviral activity, using inhibition of viral cytopathogenic effect as the primary parameter for evaluation. Inhibition of the development of intra- and extracellular virus titer was used as a secondary criterion with certain viruses. Five derivatives were considered to have significant antiviral activity. 6 Hydroxylamino-RPcMP was active against type 1 herpes simplex, cytomegalo-, and vaccinia viruses. 6-Thio-RPcMP was inhibitory to types 1 and 2 herpes simplex, cytomegalo-, vaccinia, and type 3 parainfluenza viruses. The 6-methylthio derivative was active against types 1 and 2 herpes simplex, cytomegalo-, and vaccinia viruses, and types 1A, 2, 8, and 13 rhinoviruses; alteration of this 6 substitution to 6-ethylthio or to 6-benzylthio weakened the herpes- and vaccinia virus activity of the compound, but each continued to have significant antirhinovirus activity. The effect of time of addition of 6-methylthio-RPcMP to type 1 herpes simplex virus-infected cells was determined; the compound was most active when added prior to the virus. Early removal of the compound from the infected cells markedly reduced its antiviral effectiveness. PMID- 15825421 TI - Chemotherapy of an experimental Fusobacterium (Sphaerophorus) necrophorum infection in mice. AB - An experimental animal model for testing antibiotics in vivo against Fusobacterium (Sphaerophorus) necrophorum has been developed. It incorporates the subcutaneous injection of the bacteria into mice followed by intraperitoneal administration of the antibiotic at 24, 48, 72, and 96 h. Mean effective dose values are based on the number of survivors 21 days after challenge. Tetracycline was the most effective drug tested, with a mean effective dose of 5.0 mg/kg, compared with mean effective dose values of 11.1 for clindamycin, 11.8 for penicillin-G, and 52.9 for lincomycin. PMID- 15825422 TI - Effect of penicillin on the in vivo formation of the D-alanyl-L-alanine peptide cross-linkage in cell walls of Micrococcus luteus. AB - Penicillin G was found to inhibit the formation of the d-alanyl-l-alanine cross linkage in intact cells of Micrococcus luteus. This reaction was approximately 50 fold less susceptible to penicillin than the formation of the d-alanyl-l-lysine cross-linkage in the same organism. The presence of two penicillin-susceptible transpeptidation reactions that function in the incorporation of peptidoglycan precursors into the cell wall is proposed. PMID- 15825423 TI - In vitro comparison of four aminoglycoside antibiotics: sisomicin, gentamicin, tobramycin, and BB-K8. AB - The activity of sisomicin, gentamicin, and tobramycin against 273 clinical bacterial isolates was about equal (except for greater activity of tobramycin against Pseudomonas) and was greater than that of BB-K 8 as determined by antibiotic dilution methods in agar and broth. PMID- 15825424 TI - BRL.8988 (talampicillin), a well-absorbed oral form of ampicillin. AB - The phthalidyl ester (BRL.8988, talampicillin) of ampicillin gave serum levels in man two and a half times those obtained with ampicillin itself. PMID- 15825425 TI - Iododeoxyuridine and herpesviral encephalitis: lack of inhibitory action against low-grade viral replication. AB - Equine herpesvirus 1 replicated in the brains of 2-week-old mice but did not produce fatal encephalitis; it thus simulated the majority of cases of herpes simplex encephalitis in man. This replication was not inhibited by iododeoxyuridine, although in tissue cultures the equine and human viruses were equally susceptible. The continued use of iododeoxyuridine for human encephalitis should be seriously questioned. PMID- 15825426 TI - [Soil quality assessment of forest stand in different plantation esosystems]. AB - After a clear-cutting of the first generation Cunninghamia lanceolata plantation in 1982, three plantation ecosystems, pure Michelia macclurei stand (PMS), pure Chinese-fir stand (PCS) and their mixed stand, were established in spring 1983, and their effects on soil characteristics were evaluated by measuring some soil physical, chemical, microbiological and biochemical parameters. After 20 years' plantation, all test indices showed differences among different forest management models. Both PMS and MCM had a favorable effect on soil fertility maintenance. Soil quality assessment showed that some soil functions, e.g., water availability, nutrient availability, root suitability and soil quality index were all in a moderate level under the mixed and pure PMS stands, whereas in a relatively lower level under successive PCS stand. The results also showed that there existed close correlations between soil total organic C (TOC), cation exchange capacity (CEC), microbial biomass-C (Cmic) and other soil physical, chemical and biological indices. Therefore, TOC, CEC and Cmic could be used as the indicators in assessing soil quality in this study area. In addition, there were also positive correlations between soil microbial biomass-C and TOC, soil microbial biomass-N and total N, and soil microbial biomass-P and total P in the present study. PMID- 15825427 TI - [Population structure and distribution pattern of rare plant communities in Houhe Nature Reserve]. AB - An investigation on the size structure and spatial pattern of 26 major tree species with a diameter at breast height (DBH) > or = 5 cm was made on a 1 hm2 fixed plot in the mixed evergreen and deciduous broad-leaved forest in Houhe National Nature Reserve of Central China. The results showed that 7 populations of the 26 species, i.e., Dipteronia sinensi, Aesculus wilsonii, Pterostyrax psilophyllus, Davidia involucrate, Euptelea pleiosperma, Tetracentron sinense and Cercidiphyllum japonicum, were rare endangered, species, and two of the 7 populations, T. sinense and C. japonicum, were very limited in numbers. The population structure of P. psilophyllus was in declining, and that of the rest was in growing. The spatial distribution pattern of P. psilophyllus, A. wilsonii and T. sinense was in random, and that of the others was clumped. Among the other 19 non-rare populations, the population structure of P. wilsonii was in declining, that of D. lotus, A. palmatum, A. wilsonii, A. franchetii, M. cuneifolia and H. dulcis was stable, and the rest was in growing. The spatial distribution pattern of D. lotus and H. dulcis was in random, and that of the others was clumped. On the whole, the growing populations were dominant, amounting to 61.54%, and the stable and declining populations accounted for 6.92% and 11.54%, respectively. The results of the two judging methods were consistent, showing that the distribution pattern of clumping (80.77%) was dominant, while the random and even distribution patterns were infrequent. PMID- 15825428 TI - [Biomass distribution patterns of Alnus hirsuta and Betula platyphylla-swamp ecotone communities in Changbai Mountains]. AB - In order to reveal the growth patterns of dominant tree species and the distribution patterns of community biomass along the horizontal environmental gradients or among the vertical layers of communities in Changbai Mountains, this paper studied the biomass distribution patterns of Alnus hirsuta-swamp and Betula platyphylla-swamp ecotone communities. The results showed that there were some differences in growth rate and in adaptability to habitats between A. hirsuta and B. platyphylla. In the wetland habitats of the ecotone, A. hirsuta grew 1-2 times faster than B. platyphylla, but along the gradient from swamp to forest, it grew slowly, while B. platyphylla grew fast. Therefore, A. hirsuta was a favorite tree species in wetland habitats. The distribution pattern of organ biomass was similar between A. hirsute and B. platyphylla, the trunk being 1/2, tree root 1/4, branch 1/10, bark 1.5/20, and leaf 1/20. The vertical distribution pattern of biomass was also similar between A. hirsute-swamp and B. platyphylla-swamp ecotone communities, the tree, shrub, and herbage layer accounted for 87%-90%, 7% 9%, and 2%-3%, respectively in the whole ecotone communities. The community biomass increased linearly from swamp to forest with the change of environment factors. PMID- 15825429 TI - [Position of Betula ermanii population ecotone in Changbai Mountains]. AB - The vegetation on the northern slope of Changbai Mountains forms a vertical zone due to the vertical differentiation of moisture and heat conditions. Ecotones are obviously existed between different vegetation zones, but it is difficult to decide their positions. In the area from 1400 m to 2200 m elevation, we adopted the methods of gradient sampling, fractal analysis, population pattern analysis, and interspecific competition index analysis to describe Betula ermanii population ecotone. The results showed that there was a forest ecotone between Betula ermanii and Picea-Abies forest near elevation 1650 m, and there was a forest line ecotone between Betula ermanii and alpine tundra near elevation 2080 m. PMID- 15825430 TI - [Major features of decline of Pinus sylvestris var. mongolica plantation on sandy land]. AB - In view of the decline of man-made sand-fixation forest of Pinus sylvestris var. mongolica in Zhanggutai sand land of Liaoning Province, this paper studied the major characteristics of the decline. The appearance of the declining man-made sand-fixation forest of Pinus sylvestris var. mongolica was grey green, its needle leaf was very thin, the blooming and fruiting rate was low, the average quantity of cones per tree was only 10.4-16.5, with only 6.96 g to 7.39 g per thousand seeds, and there were many empty and astringent seeds. The seasonal dynamics of nutrients in 2-year-old pine needle leaf was similar, i.e., the N and P contents decreased, while K content increased, showing that the nutrient cycle was imbalance. The chlorophyll content in 2-year-old needle leaf of declined forest was high, while that in 1-year-old healthy forest was also high but with a wide increasing range. The infected harm of shoot blight was the clearest mark to the decline of man-made sand-fixation forest of Pinus sylvestris var. mongolica. After the forest declined, the height and the DBH of the pine trees decreased evidently, and the structure of DBH distribution moved "left". The quantity of weak pine trees increased by 15.9%-27.2%, the roots decreased by 22.9%-28.9%, and the absorbing roots (diameter < 0.5 cm) decreased most seriously. PMID- 15825431 TI - [Decline regularity and causes of Pinus sylvestris var. mongolica plantation on sandy land]. AB - The study showed that the decline of Pinus sylvestris var. mongolica plantation out of its natural distribution area was a phenomenon of premature senility. The infected harm of shoot blight was the direct cause of Pinus sylvestris var. mongolica dying, but wasn't the basic reason. The decline of Pinus sylvestris var. mongolica was caused by the interaction of biological and other factors, which was "the third disease" beyond infectious and non-infectious diseases, namely, "decline disease of forest". Climate difference, improper management, unbalanced usage of soil water, and high content of N settled from air were the inducing factors, among which, climate difference and unbalanced usage of soil water were the main ones, while insect harm and shoot blight were the promoting factors. PMID- 15825432 TI - [Effects of light and temperature on seed germination of Picea asperata and Betula albo-sinensis]. AB - The effects of different light and temperature conditions on seed germination of two keystone species (Picea asperata and Betula albo-sinensis) in subalpine coniferous forests were studied by simulating environmental conditions in climatic chambers. The results indicated that the optimal temperature of seed germination for both species was 25 degrees C, with the germination rate of 88.8 +/- 8.4% and 30.8 +/- 10.4%, respectively. Through multiple comparisons with Tukey's HSD, B. albo-sinensis was more sensitive to temperature than P. asperata. Seeds of P. asperata could germinate when the temperature reached 10 degrees C, but those of B. albo-sinensis could not, which implied that the latter as a pioneer species of subalpine coniferous forest areas needed higher lowest temperature for seed germination. Day and night temperature difference (15 degreesC/10 degrees C, 20 degrees C/15 degrees C and 20 degrees C/10 degrees C) had significant effect on the germination rates of tested seeds, which was the highest under 20 degrees C/15 degrees C. A wide range of day and night temperature difference seemed to be unfavorable to the seed germination of B. albo-sinensis. Light intensity also had a marked effect on the germination rates of test seeds. Under 20% transmittance, the germination rate of P. asperata and B. albo-sinensis was the highest, being 83.0 +/- 6.6% and 29.2 +/- 5.5%, respectively, and the initiation time and regularity of germination were the best as well. Furthermore, B. albo-sinensis was less sensitive to light than P. asperata, which was determined by the life behavior of tree itself. PMID- 15825433 TI - [Effects of 6-BA and AsA on photosynthesis photoinhibition of attached poplar leaves under osmotic stress of root]. AB - In order to know more about the relationships between photosynthesis photoinhibition and reactive oxygen species metabolism, the effects of 6 benzyladenine (6-BA) and ascorbate (AsA) on net photosynthetic rate (Pn), apparent quantum yield (AQY), superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities, O2-* generation rate, and H2O2 and malondialdehyde (MDA) contents were studied with attached leaves of poplar clone seedlings under osmotic stress of root. Under osmotic stress, the photosynthesis photoinhibition of attached poplar leaves, judged by the significant decrease of Pn and AQY, was aggravated, and the balance of reactive oxygen species metabolism was destroyed. The superoxide dismutase (SOD) activity increased, but ascorbate peroxidase (APX) activity decreased. In the meantime, the O2-* generation rate and the contents of H2O2 and malondialdehyde (MDA) increased. When osmotic stressed poplar seedlings were pretreated with 6-BA and AsA, the activities of SOD and APX increased, O2-* generation rate and H2O2 and MDA contents decreased, and photosynthesis photoinhibition was alleviated. The contents of reactive oxygen species and MDA in poplar leaves were negatively correlated with net photosynthetic rate and apparent quantum yield. It's indicated that the photosynthesis photoinhibition of attached leaves of poplar clone seedlings had intrinsic relations with the accumulation of reactive oxygen species under osmotic stress of root, and the alleviation effects of 6-BA and AsA on photosynthesis photoinhibition were related to their promotion effects to the scavenging system of reactive oxygen species. PMID- 15825434 TI - [Effect of soil water potential on hydraulic parameters of Fraxinus mandshurica seedlings]. AB - In this paper, soil water potential was controlled in the ranges of 0-(-)20 kPa (W1), -20-(-)40 kPa (W2), -40-(-)60 kPa (W3), -60-(-)80 kPa (W4) and -80-(-)160 kPa (W5) by a suite of specially designed root-sphere osmotic irrigation technique, and the dynamics of transpiration, water suction, tissue water potential and hydraulic resistance of Fraxinus mandshurica seedlings grown under these soil water conditions were studied. The results indicated that under sub saturated soil water condition (W1), fine root water potential fluctuated at high levels from -0.08 MPa to -0.19 MPa, which brought about the lowest soil-root hydraulic resistance (35.52 MPa x s(-1) x m(-2) x mol(-1) in diurnal average), the highest water suction rate (4.44 mmol x m(-2) x s(-1) in average from 10:00 to 14:00), and the strongest transpiration rate (5.11 mmol x m(-2) x s(-1) in average from 10:00 to 14:00). Under field capacity (W2), soil-root hydraulic resistance increased several times above that under W1, water suction rate and transpiration rate declined significantly, while the diurnal single-peak pattern of transpiration was not altered. Under orderly increased soil water stress (W3 W5), soil-root hydraulic resistance (the average from 10:00 to 14:00) increased approximately 10-24 times above that under W1, which caused the consequent low water suction rate and low fluctuation levels of leaf water potential (-1.25 MPa (-)1.94 MPa for W5), and transpiration was seriously restricted (the transpiration rate being only 0.91-1.55 mmol x m(-2) x s(-1) at midday for W3 W5). It is concluded that soil water was not equally available to Fraxinus mandshurica seedlings in the test ranges of soil water potential, and significant water stress could occur when the soil water potential was lower than -40 kPa. PMID- 15825435 TI - [Photosynthetic characteristics of main urban forest species Syringa oblata in Shenyang]. AB - Using LI-6400 CO2 gas analysis system and by the methods of correlation analysis and multinomial regression, this paper studied the diurnal changes of Syringa oblata photosynthesis during different seasons and assessed the correlation between net photosynthetic rate and environmental factors. The results indicated that the photosynthetic rate showed a double-peak curve in spring and summer and a single peak curve in autumn. The major factors affecting the photosynthetic rate were the solar radiation and stomatal conductance in spring and summer, and the stomatal conductance and intercellular CO2 intensity in autumn. The related equation was also constructed in this paper. PMID- 15825436 TI - [Dynamics of nutrient element iron in soil-plant ecosystem of the Songnen Plain Leymus chinensis grassland]. AB - The study showed that in the soil-plant ecosystem of the Songnen Plain Leymus chinensis grassland, the contents of total and available iron in soil were relatively low, while Leymus chinensis was abundant in iron. The concentrations of soil total and available iron decreased with soil depth. Soil available iron positively correlated with soil organic matter, while negatively correlated with soil pH. The dynamics of soil total iron showed a "V" curve during the growth season, with the lowest in July, and that of soil available iron was similar to soil total iron from May to August, and gradually decreased after August. The iron concentration in Leynus chinensis changed dramatically with the trend of root > rhizome > leaf > litter > stem. The iron concentrations in leaf and stem had a fluctuant decrease, while those in root, rhizome and litter were fluctuated in "V" curve. Soil A layer enriched in a little more available iron. The iron activation with an average of 0.64% increased from May to August, and decreased after August, with the lowest in October. The shift ratio from underground to aboveground part of Leymus chinensis dropped during May-July, went up in August, and then dropped again. The average shift ratio from aboveground part to litter was 105.0%, which negatively correlated with that from underground to aboveground part. PMID- 15825437 TI - [Changes of soil chemical properties in sheep urine patches in Inner Mongolia steppe]. AB - Sheep play an important role in the nutrient cycling in extensively grazed grasslands, mainly through their annual return of around 5-25 kg N in the shape of urine. This return changes the temporal distribution of nutrients in soil and alters their bioavailability. Urine patches are the most important areas of the transformation and cycling of nutrients in the grazed grasslands. In order to make known the changes of soil chemical properties in sheep urine patches, an experiment was set up in the typical steppe area of central Inner Mongolia. The results indicated that soil pH value reached the maximum on the second day after urine excretion, and then declined. It maintained at around 6.5 from the 58th day, which was under the level of the control. Soil dissolved organic carbon (DOC) in urine patches exhibited the identical changing characteristics with soil pH, and had a significantly (P < 0.01) positive correlation with it. Soil total dissolved phosphorus (TDP) increased significantly (P < 0.05) after urine excretion, and reached the maximum at the 2nd day. The percentage of molybdate reactive phosphorus (MRP), dissolved organic phosphorus (DOP) and dissolved condensed phosphorus (DCP) showed significantly two changing phases, and there existed significantly (P < 0.05) negative correlations between MRP, DOP and DCP. Soil NH4+-N reached the maximum on the 2nd day after urine excretion and then declined, while soil NO3(-)-N increased significantly after 14 days and reached the maximum on the 21st day, showing significant "double peaks" characteristics. Soil dissolved organic nitrogen (DON) also increased significantly after urine excretion. PMID- 15825438 TI - [Changes of four common plant populations growth and their anti-oxidative enzymatic system in desertification process]. AB - This paper studied the changes of the growth and anti-oxidative enzymatic system of four common plant populations during the desertification process of sandy grassland. The results showed that in the process of desertification, the individual height and density and the density percentage of the populations all had a decreasing trend. The growth of Melilotoides ruthenica was more vigorous before moderate desertification (MD) stage, but restricted after that. In MD stage, the growth of Leymus chinensis was heavily restricted, and its individual height, density and density percentage accounted for 57.19%, 2.50% and 6.22% of those in original vegetation (OV) stage, respectively. The individual height and density of Cleistogenes squarrosa and Artemisia frigida increased in the stages of potential desertification (PD), light desertification (LD) or MD because of their phase status of dominant species and their stronger stress resistance. The SOD and POD activities of the common plant populations increased in PD and MD stages, but decreased in LD and heavy desertification (HD) stages. The CAT activity of Leymus chinensis was higher, whose response to desertification was not significant (P > 0.05), and that of Melilotoides ruthenica increased significantly in PD and HD stages (P < or = 0.01). The activities of the three anti-oxidative enzymes in the common plant populations, except the CAT activity of Melilotoides ruthenica, decreased in HD stage. The MDA content in the common populations increased firstly, then decreased, and finally increased from OV to MD stage, and had a significant difference in different desertification gradients (P < or = 0.05). Based on the integrated analysis of the ecological and physiological changes, it could be concluded that Leymus chinensis was more sensitive to desertification, while Melilotoides ruthenica had a stronger bioenergy. PMID- 15825439 TI - [Effects of grazing on architecture and small-scale pattern of grasses on artificial grassland in subtropical zone]. AB - This study was conducted on a 5-year artificial grassland in subtropical zone of South China. The main types of established artificial grassland there were Dactylis glomerata-Lolium prenne-Trifolium repens and D. glomerata-T. repens pastures. Four grazing intensities were designed, i.e., CK (no grazing), G1 (6 adult sheep x hm(-2)), G2 (7.5 adult sheep x hm(-2)) and G3 (10 adult sheep x hm( 2)), and all the grazing plots were rotationally grazed. The architecture and small-scale pattern of grasses on the grazed and ungrazed grassland were discussed. After a period of 5-year grazing, the plant basal width and sward height of D. glomerata and T. pratense decreased gradually. In treatments CK, G1, G2 and G3, the basal width of D. glomerata was 6-8, 2-4, 0-2 and 0-2 cm, and that of T. pratense was 1-1.2, 6-8, 4-6 and 2-4 cm, respectively. The tuft density of D. glomerata in treatments CK, G1, G2 and G3 was 60, 95.1, 210.2 and 160 tufts x m(-2), respectively. The tiller number per plant of D. glomerata decreased, while its tuft density increased significantly due to the increased grazing intensity. With the increase of grazing intensity, the internode length of T. repens decreased, while its branching angle increased. The average internode length in treatments CK, G1, G2 and G3 was 2.04, 1.69, 1.64 and 1.51 cm, while the branching angle was 46.5, 65, 73 and 77.3 degrees, respectively. The average leaf density of T. repens in treatments CK, G1, G2 and G3 was 2.9, 13.0, 4.7 and 1.0 x m(-2), while the relevant stolon density was 19.9, 101, 142 and 82.6 m x m(-2), respectively. Under moderate grazing intensity, both the leaf and stolon densities of T. repens increased. The main scale on small pattern of D. glomerata, T. repens and T. pratense was 2 cm x 2 cm, which was further decreased under higher grazing intensity in the treatments of D. glomerata and T. pratense. Considering the heterogeneity of environmental resources, the change of the architecture and small-scale pattern could be regarded as an adaptation of grasses under grazing disturbance. PMID- 15825440 TI - [Snow disaster in Aletai Region--its remote sensing monitoring model and evaluation method]. AB - In this paper, a monitoring model of snow depth was built based on the 4 scenes of NOAA satellite digital images under sunshiny condition and the corresponding ground observation data from 20 meteorological stations during 2 snow disasters from 1996 to 1997 in North Xinjiang. The pixel-based snow coverage rate and snow spatial classification were studied by using linear mixture spectrum disassembling method, and two grid data layers based quantified indices used for estimating snow hazard grade of grassland and animal husbandry were put forward. The results indicated that by using the snow monitoring model and linear mixture spectrum disassembling method, the image cell based snow depth and snow coverage rate could be calculated, and the precision of snow classification could be improved. The image cell based snow hazard index could systematically express the spatial distribution of snow, grass, animal and climate conditions, and reflect the snow hazard grade of grassland and animal husbandry. PMID- 15825441 TI - [Relationships between biochemical qualities of paddy rice and climate conditions in the Anning River Valley]. AB - Based on the experimental data of paddy rice planted geographically periodically without control in the Anning River Valley of Sichuan Province, this paper quantitatively analyzed the effects of meteorological conditions from 40 days before heading to ripeness on the biochemical qualities of paddy rice, and developed a model about the integrated relationships between biochemical qualities of paddy rice and mean temperature, daily range of temperature and daylight hours, which would be of significance both for the instruction of paddy distribution in the Anning River Valley and for improving rice qualities via adjusting planting time. The results showed that climate conditions had a great effect on the biochemical qualities of paddy rice, which was different in ways and in critical periods. The period when the correlation between some ingredients of rice and climate conditions was most significant was before or after, or from before to after full heading, which widened the existing knowledge about the period which was important for paddy rice qualities forming. Applying this finding in the Anning River Valley during 2002-2003, the accumulated high-grade paddy farming area stood at 73,000 hm2, 30,000 hm2 more than that in 2001, and brought 0.2 billion yuan increment to the peasants, promoting the development of high-grade paddy greatly. PMID- 15825442 TI - [Effects of small amount precision seeding on winter wheat yield]. AB - A two-year autumn seeding experiment was conducted during 2001-2003 in Beijing to study the effects of small amount precision seeding on winter wheat yield. When the seeding amount was 22.5 kg x hm(-2), the best average yield of winter wheat varieties DS No.1 and Linkang No.1 was 6836.25 and 7353.75 kg x hm(-2), respectively, and some experimental plots had a yield surpassed 7500 kg x hm(-2). The test varieties had a normal expression of growth and development in their growth period, and the contents of total saccharide, proline and lysine in seedlings were higher, and the tillering ability of plant was stronger than the control. The net photosynthesis and transpiration rates, RS, COND and CINT of flag leaf showed the vigorous physiological functions of the plants, and the higher activities of nitrate reductase and SOD showed their stronger metabolism activity. There were more spikes per plant for the test varieties. In practicing small amount precision seeding, variety selection is the prerequisite, and sowing amount is the heart of the matter. PMID- 15825443 TI - [Relationships between water-stable aggregates and nutrient status in black soil after reclamation]. AB - The study on the dynamics of water-stable aggregates in black soil after reclamation and its relation to soil nutrient status showed that the contents of soil water-stable aggregates decreased significantly at initial reclamation stage, but the declining became small with time. > 0.25 mm water-stable aggregates had a significant correlation with soil organic C, total N, total P and CEC, the correlation coefficients being 0.7625, 0.6794, 0.6084 and 0.8134, respectively, and also had a significant correlation with soil exchangeable Ca, Mg, K, Na. A closer relationship of the aggregates was found with soil organic C and CEC than with other fertility properties, while no evident correlation existed between the aggregates and soil total K, available P, available K and slow-release K. In order to maintain a good aggregate structure of black soil, it is of most importance to concern a proper balance of soil organic C. PMID- 15825444 TI - [Effects of land use and vegetation restoration on soil quality in a small catchment of the Loess Plateau]. AB - Soil quality improvement plays an important role in sustaining global biosphere. This paper studied the changes of soil quality after 25 years' land use and vegetation restoration at the Anjiapo catchment of western Loess Plateau. The analyses of soil characteristics of wasteland, almond land, farmland, pineland, shrub land and fallow land showed that different land use and vegetation restoration had different effects on soil integrated fertility index. Soil organic matter content was increased due to planting shrubs and forests. Both vegetation restoration and fallow could improve soil quality. Cultivation practice could decrease soil nutrient levels, and cropland soil was degraded. Shrub soil formed "fertile island" in the semi-arid region. Fallow could improve soil fertility to some extent. Human activities and vegetation restoration could affect soil nutrient contents after land use pattern was changed. With the launch out into the "Grand development of Western China", "Grain for Green Project" and ecological restoration, both shrub planting and fallow (natural restoration) should be the optional choices to restore soil fertility, as they could decrease soil erosion and improve soil condition at catchment scale, especially in the hilly and gully loess area. Integrative control of small catchment may be the best way for the sustainable development of the semi-arid hilly area of Loess Plateau. PMID- 15825445 TI - [Dynamic analysis of land use and land coverage in mid-eastern China during 1988 2000]. AB - Using remote sensing (RS) and geographic information system (GIS) technologies and based on the documentary and field sampling data, this paper constructed and analyzed the 1988 and 2000 landscape maps of mid-eastern China. The mapping area of mid-eastern China was about 2.87 x 10(6) km2. With the rapid increase of economy and population, the area of farmland and town in mid-eastern China increased 10.921 x 10(5) hm2 and 13.754 x 10(5) hm2, accounted for a 2.14% and 19.25% increase, respectively from 1988 to 2000. At the same time, the forest area decreased 17.83 x 10(4) hm2, declined by 1.12%, because of the increasing demand for lumber provision and reclamation. The meadow area in 2000 increased 19.287 x 10(5) hm2, ascending 7.75%, while the coverage and quality of grassland declined, owing to the deteriorating environment. The untilled land in 2000 dropped 34.31%, contrast with 1988 by the reason of a large number of lands being exploited. PMID- 15825446 TI - [Interactions between fungal-feeding nematodes and fungi and their effects on soil nitrogen mineralization]. AB - A species of fungal-feeding nematodes (Aphelenchus avenae) and two species of fungi (Micheli corticolus and Moniliaceae sp.) were isolated from an alluvial soil planted with paddy-rice and wheat in subtropical region. In order to approach their interactions and effects on soil nitrogen mineralization, a gnotobiotic microcosm experiment was carried out with four treatments, i.e., 1) soil + fungi-I (M. corticolus), 2) soil + fungi I + Aphelenchus avenae, 3) soil + fungi II (Moniliaceae sp.), and 4) soil + fungi II + Aphelenchus avenae at 22 degrees C for 25 days. The results showed that the individuals of nematode and fungi were increased simultaneously, suggesting that the inoculation of A. avenae stimulated the growth of fungi. The fungal-feeding nematode increased during the initial 20 days of incubation and then decreased slowly, and the individuals of the nematode that fed on Moniliaceae sp. (II) were higher than those fed on M. corticolus (I) (P<0.01). In the first 12 days of incubation, the population of Moniliaceae sp. (II) increased 2.5-3.5 times, while that of M. corticolus (I) increased slower but caught up after the 12th day, and the density reached 5.0 5.7 times higher than the corresponding treatments with only fungi inoculation. The interactions encouraged the soil nitrogen mineralization of all four treatments, as compared with the control. Comparing the treatments with and without fungal-feeding nematode inoculation, the treatments with fungal-feeding nematode had a higher content of soil NH4+-N than those with only fungi inoculations, which indicated that nematode had a greater effect on soil nitrogen mineralization than fungi. As for the two species of fungi, M. corticolus (I) had a greater effect on soil nitrogen mineralization than Moniliaceae sp. (II). It was observed, however, that the nitrification rate declined as compared with the control. PMID- 15825447 TI - [Effects of Bacillus thuringiensis transgenic rice and chemical insecticides on arthropod communities in paddy-fields]. AB - This paper assessed the effects of cry1Abl/cry1Ac-carrying transgenic Bacillus thuringiensis rice (Bt rice) and chemical insecticides on the arthropod communities under paddy-field conditions in terms of arthropod guild dominance, family composition and dominance distribution of each guild, individuals of each guild, some common indices of arthropod communities, and community dissimilarity. In most cases, no significant differences were found between the Bt and control rice plots without any chemical insecticide spray in such arthropod community specific parameters as guild dominance, family composition and dominance distribution, temporal dynamics of the individuals of each guild or all arthropod, as well as some common community indices (including species richness, Shannon-Wiener diversity index, evenness index and dominant index) and their temporal dynamics. The similarity of arthropod community between the Bt and control rice plots were apparently higher. It was clear that Bt rice generally showed no marked negative effect on the arthropod community in paddy field. In contrast, some arthropod community specific parameters such as guild dominances in the control plot with chemical insecticide spray were in some cases markedly and even significantly different from those in the control plot without any chemical insecticide spray. The similarity of arthropod community between the control plots with and without chemical insecticide spray was relatively lower. It could be concluded that the effect of Bt rice on the arthropod community was apparently lower than that of chemical insecticides. PMID- 15825448 TI - [Host transfer of Beauveria bassiana population in pine stand ecosystem and impact of its genetic diversity on sustainable control of Masson's pine caterpillars]. AB - In an anniversary investigation post inoculative release of Beauveria bassiana against Masson's pine caterpillars in a Masson's pine plantation, 119 strains were isolated from 30 host insects, soil, litter and air. Based on an esterase isozyme analysis, all these strains were assigned to 32 esterase types, suggesting abundant genetic diversity. The esterase type 2 was assigned including 18 other strains from 11 different host insects, indicating that the released strain had successfully survived in different insect populations and persisted in enzootic condition in 12 insect populations including pine caterpillars and grey pine weevils. Other host insects could maintain the food chain when the target insect population was low. The other esterase types included 1-23 strains. The anniversary host transfer dynamics further revealed that B. bassiana could transfer among different host insects in the pine ecosystem. The virulence of the strains of each esterase differed substantially, which suggested that B. bassiana persisted and dispersed along more than one route, and each esterase stood for at least one branch in the food web. Some hosts connected different esterase types at nodal points, making the food web very complicated. In addition, the strains isolated from soil, litter, forest crown and air belonged to different esterase types, suggesting that a complex of saprophytic food chains existed in the plantation, which made the food web more complicated, and benefited the sustainable control of the Masson's pine caterpillar and other pest insects in the ecosystem. PMID- 15825449 TI - [Effects of ultra low volume Beauveria bassiana oil formulation on biological diversity]. AB - In 2001-2003, an ultra low volume (ULV) Beauveria bassiana (Bb) oil formulation was used to prevent against Dendrolimus punctatus, and its effect on biological diversity (species richness, individual number, species diversity index and evenness index, and dominant index) was investigated. The results showed that there were 473, 392 and 266 species in non-control, Bb control and chemical control areas, respectively. The individual number ratio of natural enemies to pest in Bb control area (0.1049:1) was close to that in non-control area (0.1051:1), but much higher than that in chemical control area (0.0558:1). In Bb- and chemical control areas, the species richness and diversity decreased, but dominant index increased. There existed significant difference between chemical control and non-control areas, but no significant difference was found between Bb control and non-control areas. ULV Bb had no obvious effect on the natural enemies and non-target insects. PMID- 15825450 TI - [Relationships between Aphis gossypii and its natural enemies in megranate field]. AB - In this paper, the relevancy of Aphis gossypii with its natural enemies was studied by using grey systematic analysis. The results showed that the major natural enemies of Aphis gossypii were Tetragnathidae (0.8607), Therdion octomaculatum (0.8058), Therdion octomaculatum (0.7989), Haramonia aiyridis (0.7881) and Chrysopa sepetempunctata (0.7758). As for the daily total predation, the natural enemies which highly associated with the ideal dominant natural enemies were Erigonidium graminicola (0.8975), Tromobidiidae (0.8132), Propylacea japonica (0.7806) and Chrysopa sepetempunctata (0.7669); while for the quantity, Erigonidium graminicola (0.8482), Chrysopa sinica (0.7533), Tetragnathidae (0.7532), Therdion octomaculatum (0.7411) and Chrysopa sepetempunctata (0.7716) were the major natural enemies. According to the relational grade of the individuals between various natural enemies and ideal dominant natural enemies, the major natural enemies were Erigonidium graminicola (0.8461), Tromobidiidae (0.7325), Propylacea japonica (0.6983), Chrysopa sepetempunctata (0.6815) and Chrysopa sinica (0.6757). The species having >0.9567 horizontal and vertical niche overlaps with Aphis gossyppii were Erigonidium graminicola, Chrysopa sepetempunctata and Tetragnathidae, while the temporal niche overlaps between Erigonidium graminicola, Chrysopa sepetempunctata, tetragnathidae and Aphis gossyppii were >0.4020. It could be concluded that the main natural enemies of Aphis gossyppii were Erigonidium graminicola, Chrysopa sepetempunctata and Tetragnathidae. PMID- 15825451 TI - [Impact of automobile exhaust on membrane lipid peroxidation and protective enzyme activities in seedlings foliage of four northern broadleaved tree species]. AB - By means of fumigating one-year-old seedlings in open top chambers, this paper studied the impact of automobile exhaust on the pH value, relative conductivity, malondialdehyde (MDA) and chlorophyll contents, superoxide dismutase (SOD) and peroxidase (POD) activities, and ascorbic acid (ASA) content in the seedlings foliage of four tree species, Acer mono, Malus baccata, Prunus ussuriensis, and Acer ginnala. During the fumigation, the seedlings were exposed to the same exhaust gas concentration (25 microg x m(-3), indicated by the NO2 concentration in exhaust) for different durations (1, 3, 5, 7 d), and to different concentrations (40, 60, 80, 100 microg NO2 x m(-3)) for same duration (2 h). The results showed that the pH value and the chlorophyll and ascorbic acid (ASA) contents decreased, whereas the relative conductivity, malondialdehyde (MDA) content, and superoxide dismutase (SOD) and peroxidase (POD) activities increased with increasing fumigation duration and exhaust concentration. Obvious interspecies variations in term of physiological features were found. After treated 7 days with 25 microg NO2 x m(-3) and treated 2 h with 100 microg NO2 x m(-3), only a 1.5% and 2.7% decrease of cell juice pH was found in A. ginnala, respectively, compared to the control. The corresponding data for P. ussuriensis was 9.42% and 13.89%, followed by M. baccata. The chlorophyll content of A. mono, A. ginnala, M. baccata and P. ussuriensis was 83.0%, 71.3%, 68.7% and 54.9%, respectively of the control after 7 days treated with 25 microg NO2 x m(-3), and the corresponding data under 100 microg NO2 x m(-3) treatment was 60.2%, 73.1%, 43.4% and 51.2%, respectively. The decrease of ASA content and Acer ginnala was less in A. mono than in M. baccata and P. ussuriensis. The relative conductivity and MDA content of A. mono increased respectively by 68.1% and 52.5% in compared with control, while those of A. ginnala had the least increment. As for the 100 microg NO2 x m(-3) treatment, the maximum increases of relative conductivity (99.8%) and MDA content (52.5%) were found in M. baccata, while the least were found in A. mono and A. ginnala. The SOD and POD activities generally increased under the highest concentration of exhaust, with the exception of M. baccata. To summarize, almost all the test physiological features of A. ginnala were least affected, suggesting its great tolerance to exhaust stress, but in contrary, the greatest variation of the physiological features and the dramatic decline of SOD and POD activities of M. baccata at the highest exhaust concentration indicated that this tree species had the least tolerance to exhaust stress. PMID- 15825452 TI - [Purification efficiency of several wetland macrophytes on COD and nitrogen removal from domestic sewage]. AB - In order to investigate the role of wetland macrophytes in waster water purification and to select appropriate native filter plants in constructed wetland, three vertical-flow constructed wetlands were built with river sands as the substrates of Acorus gramineus, Juncus effusus and Iris japonica, and one without plant as the control. Investigation on the removal of COD and total nitrogen (TN) from domestic sewage showed that within lower concentrations of COD (<200 mg x L(-1)) and TN (<30 mg x L(-1)), more than 90% of COD and 80% of TN were removed from domestic sewage in all constructed wetlands. When the concentration of COD and TN increased, the purification efficiency of all constructed wetlands decreased to some extent. The constructed wetlands with macrophytes had a higher efficiency than control. Among the three constructed wetlands with macrophytes, the one with Acorus gramineus had an average purification efficiency of 80.46% for COD and 77.77% for TN, that with Juncus effusus was 75.53% for COD and 71.17% for TN, and the one with Iris japonica was 70.50% for COD and 66.38% for TN. The constructed wetland without vegetation had an average purification efficiency of 61.39% for COD and 55.81% for TN. Acorus gramineus was more capable of removing COD and TN than Juncus effusus and Iris japonica. Vegetation biomass was the main factor affecting the removal rate of COD and nitrogen, because it significantly correlated with the ability of absorbing organic substance and nitrogen, and with the nitrification and denitrification around roots. PMID- 15825453 TI - [Weed-suppressive effect of phenolic acids]. AB - Allelochemical substitutions salicylic acid, p-hydroxybenzonic acid, cinnamic acid, vanillic acid and ferulic acid were tested in a gyrating regression experiment to study the weed-suppressive effect of phenolic acids on the root growth of barnyardgrass. The results showed that cinnamic acid had a significant inhibitory effect, its square item regression coefficient being -6.18; salicylic acid, p-hydroxybenzonic acid and ferulic acid had the same trend of weed suppressive potential as cinnamic acid, while vanillic acid had a different weed suppressive effect from the others. The inhibition rate (IR) on the root growth of barnyardgrass reached its highest level (78.5%) when 0.06 mmol x L(-1) salicylic acid, 0.60 mmol x L(-1) p-hydrobenzonic acid, 0.24 mmol x L(-1) cinnamic acid, 0.02 mmol x L(-1) vannillic acid and 0.02 mmol x L(-1) ferullic acid were mixed together to apply. PMID- 15825455 TI - [Heavy metals accumulation in different parts of Paeonia ostii and soils at copper tailings yard]. AB - The study on the heavy metals contamination of Paeonia ostii and soil at the copper tailings yard in Tongling City indicated that the soil of copper tailings yard was extremely barren, only with 1.1-3.4 g x kg(-1) organic mater (1/15 of normal soil), but the contents of its Cu, Cd, Pb and Zn were much higher. Cu and Cd contents were 587.43-1176.44 mg x kg(-1) and 3.08-5.16 mg x kg(-1), respectively, 5-10 times higher than the grade two of national environmental quality standard for soils. Cu, Cd and Pb contents in Paeonia ostii exceeded the standard of medicine-use plants. Especially in its root peel, Cu and Cd contents stood at 31.50-64.00 mg x kg(-1) and 0.98-1.45 mg x kg(-1), respectively, 1.6-3.6 times surpassing the standard, which meant that both Paeonia ostii and the soil had been severely polluted. There existed evident difference in the contents of heavy metals in different parts of Paeonia ostii. Stem, leaf and root peel had higher contents of Zn, Cd, Pb and Cu. The accumulation factors of Zn and Cd were higher than those of Cu and Pb, but they were all smaller in root peel. PMID- 15825454 TI - [Mitigation effect of several controlled-release N fertilizers on ammonia volatilization and related affecting factors]. AB - By using static absorption and soil column leaching methods, this paper studied the behaviors of several controlled-release N fertilizers in soil under laboratory conditions. The results showed that under the application rate of 450 mg x kg(-1), total ammonia volatilization from three controlled-release fertilizers decreased by 49.7%, 28.0% and 71.2%, respectively, in comparing with common urea. When the application rate was 600 mg x kg(-1), total ammonia volatilization decreased by 34.6%, 12.3%, 69.9%, respectively. Controlled-release fertilizers could markedly reduce total ammonia volatilization from soil and decrease environment pollution via fertilization. The results also indicated that total ammonia volatilization correlated significantly with soil urease activity, pH value and N leaching rate. The correlation coefficient between total ammonia volatilization and accumulated N leaching rate was 0.9533, and that between total ammonia volatilization and soil urease activity and pH value was 0.9533 and 0.9908, respectively. PMID- 15825456 TI - [Cu and Pb contents in Dichondra repens leaf and their effects on its physiological indexes]. AB - This paper analyzed the contents of Cu and Pb in Dichondra repens grown on Cu and Pb contaminated soil to determine if this plant is a hyperaccumulator of two elements. The results indicated that total content of Cu (44.8 g x kg(-1)) in the plant was higher than that of Pb (25.59 g x kg(-1)), and the contents of two elements were higher in roots and stems than in leaves and increased with increasing soil contamination. The accumulation coefficient of Cu (0.784) was higher than that of Pb (0.465). With the increasing concentrations of two heavy metals, the chlorophyll a content and chlorophyll a/b ratio in leaf decreased, the SOD activity increased to the peak and decreased subsequently, while the POD activity always maintained an increasing trend. The membrane permeability of cell was not significantly varied. PMID- 15825457 TI - [Using paper as the carbon source for groundwater nitrate removal]. AB - The nitrate removal from groundwater was studied in laboratory columns packed with unprocessed paper as the sole substrate for microbial population. The results showed that the reactor with paper as carbon source started up fast, and temperature and hydraulic retention time (HRT) markedly affected the denitrification process. The denitrification rate at 25 degrees C was approximately 1.7 times higher than that at 14 degrees C. When the initial concentration was 45.2 mg x L(-1), 99.6% of nitrate (nitrite was not detected) could be removed after 8.6 h at 25 +/- 1 degrees C. The nitrogen removal rate was only 50% when HRT was 7.2 h. pH and dissolved oxygen (DO) had little effect on denitrification, and the nitrogen removal rate did not changed when pH value ranged between 6.0-9.0 and DO ranged between 0-6 mg x L(-1). Biofilm was formed on the paper and consumed the paper during the whole process. Using the combined paper reactor and activated carbon adsorption at 25 +/- 1 degrees C and HRT 9.0 h, the nitrate could be completely removed if its initial concentration was not higher than 33.9 mgN x L(-1) and the dissolved organic carbon (DOC) in the effluent was less than 11 mg x L(-1). PMID- 15825459 TI - [Research advances in dynamic mechanism and its simulation of eco-hydrological process in forest catchment]. AB - Hydrological process is the key link between climatic fluctuations and ecologic dynamics of forests at spatial and temporal scales. To some extent, global climate change and large-scale human's activities will impact the spatial temporal changes of eco-hydrological process in forest catchments in the future. Therefore, researches on the dynamic mechanism of eco-hydrological processes in forest catchments play a crucial role in understanding and controlling the rational use of ecological resources, resuming regional ecology, and sustainable development in economy. This paper described the interception, evapotranspiration and rainfall-runoff of forest ecological system and their effects on hydrological process. The spatial structure of soil moisture and its evolution with time were also the cause and consequence of forest. The spatial-temporal interaction between hydrologic and ecologic dynamics, the application of distributed hydrological model, and the eco-hydrological dynamics and cybernetics of forest would be the most exciting frontiers of the relative researches in the future. PMID- 15825458 TI - [Ecosystem approach: a new concept for ecosystem management]. AB - Ecosystem approach (EA) was firstly proposed by ecologists in developed countries and then supported by a number of international institutions and NGOs, among which, CBD, IUCN and WWF played important roles. Ecosystem approach is an integrated strategy for the management of land, soil and bio-resources. The application of EA will help to reach a balance between conservation, sustainable use, and fair and equitable sharing of the benefits arising out of the utilization of biological resources. Ecosystem approach is a methodology of ecosystem management, focusing on the biological organisms and recognising that human beings, with their cultural diversity, are an integral component of many ecosystems. The decision V/6 adopted by the Conference of the Parties to the CBD at its fifth meeting in 2000 concretized the EA in the form of twelve principles and five operational guidelines. Our government has recently made a lot of efforts in ecosystem management at large scale with many important measures and obtained significant achievements, but the potential role of local governments, institutions and individuals has not been fully played. In the attempts of managing a specific ecosystem, there are many successful cases done by our ecologists, for example, the ecological management of Chinese fir plantation forest in central subtropical China. However, the whole ecosystem at national or regional level is confronted with a lot of serious problems, mainly because there is a lack of complete understanding of the significance of ecosystem management and a lack of guidelines or principles from an integrated scientific theory. The introduction and implementation of ecosystem approach will play an important role in improving the ecosystem management in China. PMID- 15825460 TI - [Organic agriculture and sustainable development]. AB - Basing on the research and practice of organic agriculture at home and abroad, this paper discussed the objectives of developing green food and the principles that must be persisted in the practice in China. In the light of the arguments concerning with sustainable agriculture, we also discussed the significance of "alternative agriculture" in theory and practice. Compared with conventional high intensity agriculture, the production approaches of organic alternatives can improve soil fertility and have fewer detrimental effects on the environment. It is unclear whether conventional agriculture can be sustained because of the shortcomings presented in this paper, and it has taken scientists approximately one century to research and practice organic farming as a representative of alternative agriculture. The development of green food in China has only gone through more than ten years, and there would be some practical and theoretical effects on the development of China's green food if we exploit an environment friendly production pattern of organic agriculture which majors in keeping human health and maintaining sustainable agriculture. PMID- 15825461 TI - [Physiological and biochemical bases of xero-tolerance of fungal biocontrol agents and their application]. AB - Hyphomycetes fungal biocontrol agents usually infect host insects by conidial germination and penetration into insect integument under high humidity condition which is generally the restrictive factor for the application of mycoinsecticides in insect control in the field. It is ideal to produce more xero-tolerant inocula (e.g. conidia) of the agents, although the current formulation technology has helped to some degree to reduce the dependence of fungal formulations on the high humidity. This paper reviewed the advance in basic and applied studies on the physiological and biochemical bases of fungal xero-tolerance and its water activity regulation in the past two decades. Possible approaches to make fungal formulations more xero-tolerant were discussed, with an emphasis on the increased accumulation of biologically compatible solutes such as low-molecular polyols and trehalose to the inocula produced for formulation. More efforts are necessitated to understand the mechanisms involved in molecular biology and in regulating the accumulation of the compatible solutes, and to explore for new technology to enhance the xero-tolerance of the fungal agents through biochemical regulation in mass production. PMID- 15825462 TI - [Influence of elevated atmospheric CO2 on rhizosphere microbes and arbuscular mycorrhizae]. AB - The changes of microbial communities in rhizosphere and the formation of mycorrhizae play an important role in affecting the dynamics of plant communities and terrestrial ecosystems. This paper summarized and discussed the effects of elevated atmospheric CO2 on them. Under elevated atmospheric CO2, the carbohydrates accumulated in root systems increased, and the rhizospheric environment and its microbial communities as well as the formation of mycorrhizae changed. It is suggested that the researches in the future should be focused on the effects of rhizosphere microbes and arbuscular mycorrhizae on regulating the carbon dynamics of plant communities and terrestrial ecosystems under elevated atmospheric CO2. PMID- 15825463 TI - [Comparison of clonal architecture between two divergent Leymus chinensis types in Songnen grassland]. AB - This paper studied the clonal architecture of two divergent Leymus chinensis types (grey-green type and yellow-green type) in Songnen grassland, and compared their internode length, spacer length, interbranching length, interbranching angle, and ramet population density and height under the same habitat. The results showed that there was no significant difference in these clonal characteristics except spacer length and ramet population density between the two types of L. chinensis, and yellow-green type, with less spacer length and more ramet density than grey-green type, should be more adaptable to the resourceful habitat. Moreover, the V-indices of the clonal architecture of two divergent L. chinensis types were all close to 1, and the difference was not significant. Therefore, both of the two types belonged to typical guerilla clonal plant. PMID- 15825464 TI - [Effects of soil salt and water contents on growth and biomass allocation of Taxodium distichum]. AB - One year old Taxodium distichum seedlings were grown in pots with varied soil water and salt (NaCl) levels (alone and in combination) under greenhouse conditions to determine their effects on the growth and biomass allocation of the seedlings. The results showed that there were significant effects of soil salt and water contents on the relative height growth, relative stem diameter growth, biomass increment and allocation, shoot/root ratio, seedling survival, ratio of branch to stem area, ratio of leaf mass to stem area, and ratio of leaf mass to branch area. According to the biomass increment, relative height growth and relative stem diameter growth, 20 treatments were classified into 3 groups by means of systematic cluster analysis. The first one was the treatments waterlogging, flooding, waterlogging + 0.15% NaCl, flooding + 0.15% NaCl, and waterlogging + 0.3% NaCl, which had a high growth, the second one was the treatments flooding + 0.45% NaCl, flooding + 0.3% NaCl, 75% of field capacity, and 75% of field capacity+ 0.15% NaCl, which had a medium growth, and the last one was the treatments 50% of field capacity, 50% of field capacity + 0.15% NaCl, waterlogging + 0.45% NaCl, 25% of field capacity, 25% of field capacity + 0.15% NaCl, 75% of field capacity + 0.3% NaCl, 75% of field capacity + 0.45% NaCl, 25% of field capacity + 0.3% NaCl, 50% of field capacity + 0.3% NaCl, 50% of field capacity + 0.45% NaCl, and 25% of field capacity + 0.45% NaCl, which had a poor growth. PMID- 15825465 TI - [Interspecific competition between Trichogramma confusum and T. pretiosum on Corcyra cephaloica factitious eggs]. AB - By means of single and mixed inoculation, this paper studied the interspecific competition between T. confusum and T. pretiosum on the factitious eggs of Corcyra cephaloica under different parasitoid densities, host densities and inoculated spaces. The results showed that for both singly and mixed inoculated groups, the parasitism increased with parasitoid density but decreased with host density, whereas the percentage of female progeny dropped with parasitoid density but enhanced with host density. No significant effect was observed on adult emergence for all parasitoid and host density treatments. In mixed inoculated group, the proportion of T. pretiosum in the progeny decreased with parasitoid density but increased with host density, and was more than 50% in all treatments, indicating that T. pretiosum had a stronger competitive ability than T. confusum. In the spaces ranging from 4 cm3 to 102 cm3, the parasitism decreased gradually in both singly and mixed inoculated groups. The percentage of female progeny and adult emergence had no significant difference among different inoculated spaces. In mixed inoculated group, the proportion of T. pretiosum in the progeny was more than 50% in all treatments except space 102 cm3, but decreased with space, which suggested that T. confusum could improve their competitive ability through increasing their search areas and looking for more hosts. PMID- 15825466 TI - What is in a name? PMID- 15825467 TI - Evidence shows changing roles of health care risk managers. PMID- 15825468 TI - Misuse of forced-air warming devices can be hazardous. PMID- 15825469 TI - Learning from near misses through reflection: a new risk management strategy. PMID- 15825470 TI - An integrated delivery system's approach to identifying and reducing risk. PMID- 15825471 TI - Criminal background checks help ensure patient safety. PMID- 15825472 TI - Emergency medicine. New Mexico state law partially preempted by EMTALA. Godwin v. Memorial Medical Center. PMID- 15825473 TI - Medical malpractice. States differ on tolling of statute in cases involving minors. Piselli v. 75th St. Medical Center; Seamans v. MacMillan. PMID- 15825474 TI - Medical malpractice/products liability. Learned intermediary rule will protect device manufacturer. Ellis v. C. R. Bard, Inc. PMID- 15825475 TI - Error concealment for shape in MPEG-4 object-based video coding. AB - In asynchronus transfer mode networks, cell loss or channel errors can cause data to be dropped in the channel. When digital images/videos are transmitted over these networks, one must be able to reconstruct the missing data so that the impact of the errors is minimized. In this paper, we present an error-concealment technique for shape in MPEG-4 object-based video coding. This method, which is based on using global motion estimation and compensation techniques for boundary recovery, consists of three steps: (1) boundary extraction from shape; (2) boundary patching using global motion compensation; and (3) boundary filling to reconstruct the shape of the damaged video object planes. Global motion parameters are inserted as part of the USER_DATA field in the compressed stream and are utilized in reconstructing the damaged boundaries of compressed video object planes. PMID- 15825476 TI - Dynamic contrast-based quantization for lossy wavelet image compression. AB - This paper presents a contrast-based quantization strategy for use in lossy wavelet image compression that attempts to preserve visual quality at any bit rate. Based on the results of recent psychophysical experiments using near threshold and suprathreshold wavelet subband quantization distortions presented against natural-image backgrounds, subbands are quantized such that the distortions in the reconstructed image exhibit root-mean-squared contrasts selected based on image, subband, and display characteristics and on a measure of total visual distortion so as to preserve the visual system's ability to integrate edge structure across scale space. Within a single, unified framework, the proposed contrast-based strategy yields images which are competitive in visual quality with results from current visually lossless approaches at high bit rates and which demonstrate improved visual quality over current visually lossy approaches at low bit rates. This strategy operates in the context of both nonembedded and embedded quantization, the latter of which yields a highly scalable codestream which attempts to maintain visual quality at all bit rates; a specific application of the proposed algorithm to JPEG-2000 is presented. PMID- 15825477 TI - Mutual information-based analysis of JPEG2000 contexts. AB - Context-based arithmetic coding has been widely adopted in image and video compression and is a key component of the new JPEG2000 image compression standard. In this paper, the contexts used in JPEG2000 are analyzed using the mutual information, which is closely related to the compression performance. We first show that, when combining the contexts, the mutual information between the contexts and the encoded data will decrease unless the conditional probability distributions of the combined contexts are the same. Given I, the initial number of contexts, and F, the final desired number of contexts, there are S(I, F) possible context classification schemes where S(I, F) is called the Stirling number of the second kind. The optimal classification scheme is the one that gives the maximum mutual information. Instead of using an exhaustive search, the optimal classification scheme can be obtained through a modified generalized Lloyd algorithm with the relative entropy as the distortion metric. For binary arithmetic coding, the search complexity can be reduced by using dynamic programming. Our experimental results show that the JPEG2000 contexts capture the correlations among the wavelet coefficients very well. At the same time, the number of contexts used as part of the standard can be reduced without loss in the coding performance. PMID- 15825478 TI - Sparse geometric image representations with bandelets. AB - This paper introduces a new class of bases, called bandelet bases, which decompose the image along multiscale vectors that are elongated in the direction of a geometric flow. This geometric flow indicates directions in which the image gray levels have regular variations. The image decomposition in a bandelet basis is implemented with a fast subband-filtering algorithm. Bandelet bases lead to optimal approximation rates for geometrically regular images. For image compression and noise removal applications, the geometric flow is optimized with fast algorithms so that the resulting bandelet basis produces minimum distortion. Comparisons are made with wavelet image compression and noise-removal algorithms. PMID- 15825479 TI - Linear demosaicing inspired by the human visual system. AB - There is an analogy between single-chip color cameras and the human visual system in that these two systems acquire only one limited wavelength sensitivity band per spatial location. We have exploited this analogy, defining a model that characterizes a one-color per spatial position image as a coding into luminance and chrominance of the corresponding three colors per spatial position image. Luminance is defined with full spatial resolution while chrominance contains subsampled opponent colors. Moreover, luminance and chrominance follow a particular arrangement in the Fourier domain, allowing for demosaicing by spatial frequency filtering. This model shows that visual artifacts after demosaicing are due to aliasing between luminance and chrominance and could be solved using a preprocessing filter. This approach also gives new insights for the representation of single-color per spatial location images and enables formal and controllable procedures to design demosaicing algorithms that perform well compared to concurrent approaches, as demonstrated by experiments. PMID- 15825480 TI - Variational image reconstruction from arbitrarily spaced samples: a fast multiresolution spline solution. AB - We propose a novel method for image reconstruction from nonuniform samples with no constraints on their locations. We adopt a variational approach where the reconstruction is formulated as the minimizer of a cost that is a weighted sum of two terms: (1) the sum of squared errors at the specified points and (2) a quadratic functional that penalizes the lack of smoothness. We search for a solution that is a uniform spline and show how it can be determined by solving a large, sparse system of linear equations. We interpret the solution of our approach as an approximation of the analytical solution that involves radial basis functions and demonstrate the computational advantages of our approach. Using the two-scale relation for B-splines, we derive an algebraic relation that links together the linear systems of equations specifying reconstructions at different levels of resolution. We use this relation to develop a fast multigrid algorithm. We demonstrate the effectiveness of our approach on some image reconstruction examples. PMID- 15825481 TI - Recovery of image blocks using the method of alternating projections. AB - A technique for block-loss restoration in block-based image and video coding, dubbed recovery of image blocks using the method of alternating projections (RIBMAP), is developed. The algorithm is based on orthogonal projections onto constraint sets in a Hilbert space. For the recovery of a linear dimension N size block, a total of 8N vectors are extracted from the surrounding area of an N x N missing block. These vectors form a library from which the best matching spatial information for the missing block is extracted. Recovery vectors, including both undamaged and restored damaged pixels, are introduced. The vectors are used to find highly correlated information relating to the lost pixels. To assure continuity with the surrounding undamaged area, three additional convex constraints are formulated. Adherance to these sets is imposed using alternating projections. Simulation results using orthogonal projections demonstrate that RIBMAP recovers spatial structure faithfully. Simulation comparisons with other procedures are presented: Ancis and Giusto's hybrid edge-based average-median interpolation technique, Sun and Kwok's projections onto convex sets-based method, Hemami and Meng's interblock correlation interpolation approach, Shirani et al.'s modified interblock correlation interpolation scheme, and Alkachouh and Bellanger's fast discrete cosine transformation-based spatial domain interpolation algorithm. Characteristic of the results are those of the "Lena" JPEG image when one fourth of periodically spaced blocks in the image have errors. The peak signal-to-noise ratio of the restored image is 28.68, 29.99, 31.86, 31.69, 31.57, and 34.65 dB using that of Ancis and Giusto, Sun and Kwok, Hemami and Meng, Shirani et al., Alkachouh and Bellanger, and RIPMAP, respectively. PMID- 15825482 TI - An automatic nonrigid registration for stained histological sections. AB - Automatic computer-based analyses of histological sections which are differently stained require that they are related to each other. Most registration methods are only able to perform rigid-body motion and are sensitive to noise and artifacts. Histological images, however, are accompanied by several artifacts and different contrasts, which require a nonrigid registration. In this paper, we present a hierarchical nonrigid registration algorithm able to align images, which contain minor image artifacts. The algorithm requires no a priori knowledge of the true image. The hierarchical design of the algorithm enhances robustness and accuracy, and saves computational costs. The proposed algorithm is decomposed into a fast, coarse, rigid registration step and a slower, but finer, nonrigid step. For the coarse registration, we use image pyramids, while for the second step, we combine a point-based registration with an elastic thin-plate spline interpolation. Accuracy tests, performed for 20 histological images obtained from human arteries, have shown that the error measure is acceptable, and that the image noise does not cause a problem. The associated convergence rate of the mean pixel displacement error during the rigid and nonrigid registrations is satisfying. The algorithm can be applied to various multicontrast elastic registration problems in medical imaging and may be extended to three dimensions. PMID- 15825483 TI - JPEG2000-coded image error concealment exploiting convex sets projections. AB - Transmission errors in JPEG2000 can be grouped into three main classes, depending on the affected area: LL, high frequencies at the lower decomposition levels, and high frequencies at the higher decomposition levels. The first type of errors are the most annoying but can be concealed exploiting the signal spatial correlation like in a number of techniques proposed in the past; the second are less annoying but more difficult to address; the latter are often imperceptible. In this paper, we address the problem of concealing the second class or errors when high bit planes are damaged by proposing a new approach based on the theory of projections onto convex sets. Accordingly, the error effects are masked by iteratively applying two procedures: low-pass (LP) filtering in the spatial domain and restoration of the uncorrupted wavelet coefficients in the transform domain. It has been observed that a uniform LP filtering brought to some undesired side effects that negatively compensated the advantages. This problem has been overcome by applying an adaptive solution, which exploits an edge map to choose the optimal filter mask size. Simulation results demonstrated the efficiency of the proposed approach. PMID- 15825484 TI - An orthogonal family of quincunx wavelets with continuously adjustable order. AB - We present a new family of two-dimensional and three-dimensional orthogonal wavelets which uses quincunx sampling. The orthogonal refinement filters have a simple analytical expression in the Fourier domain as a function of the order lamda, which may be noninteger. We can also prove that they yield wavelet bases of L2(R2) for any lambda > 0. The wavelets are fractional in the sense that the approximation error at a given scale a decays like O(a(lamda)); they also essentially behave like fractional derivative operators. To make our construction practical, we propose a fast Fourier transform-based implementation that turns out to be surprisingly fast. In fact, our method is almost as efficient as the standard Mallat algorithm for separable wavelets. PMID- 15825485 TI - A memory learning framework for effective image retrieval. AB - Most current content-based image retrieval systems are still incapable of providing users with their desired results. The major difficulty lies in the gap between low-level image features and high-level image semantics. To address the problem, this study reports a framework for effective image retrieval by employing a novel idea of memory learning. It forms a knowledge memory model to store the semantic information by simply accumulating user-provided interactions. A learning strategy is then applied to predict the semantic relationships among images according to the memorized knowledge. Image queries are finally performed based on a seamless combination of low-level features and learned semantics. One important advantage of our framework is its ability to efficiently annotate images and also propagate the keyword annotation from the labeled images to unlabeled images. The presented algorithm has been integrated into a practical image retrieval system. Experiments on a collection of 10,000 general-purpose images demonstrate the effectiveness of the proposed framework. PMID- 15825486 TI - Myocardial motion analysis from B-mode echocardiograms. AB - The quantitative assessment of cardiac motion is a fundamental concept to evaluate ventricular malfunction. We present a new optical-flow-based method for estimating heart motion from two-dimensional echocardiographic sequences. To account for typical heart motions, such as contraction/expansion and shear, we analyze the images locally by using a local-affine model for the velocity in space and a linear model in time. The regional motion parameters are estimated in the least-squares sense inside a sliding spatiotemporal B-spline window. Robustness and spatial adaptability is achieved by estimating the model parameters at multiple scales within a coarse-to-fine multiresoluion framework. We use a wavelet-like algorithm for computing B-spline-weighted inner products and moments at dyadic scales to increase computational efficiency. In order to characterize myocardial contractility and to simplify the detection of myocardial dysfunction, the radial component of the velocity with respect to a reference point is color coded and visualized inside a time-varying region of interest. The algorithm was first validated on synthetic data sets that simulate a beating heart with a speckle-like appearance of echocardiograms. The ability to estimate motion from real ultrasound sequences was demonstrated by a rotating phantom experiment. The method was also applied to a set of in vivo echocardiograms from an animal study. Motion estimation results were in good agreement with the expert echocardiographic reading. PMID- 15825487 TI - Enhanced resolution in Sar/ISAR imaging using iterative sidelobe apodization. AB - Resolution enhancement techniques in radar imaging have attracted considerable interest in recent years. In this work, we develop an iterative sidelobe apodization technique and investigate its applications to synthetic aperture radar (SAR) and inverse SAR (ISAR) image processing. A modified noninteger Nyquist spatially variant apodization (SVA) formulation is proposed, which is applicable to direct iterative image sidelobe apodization without using computationally intensive upsampling interpolation. A refined iterative sidelobe apodization procedure is then developed for image-resolution enhancement. Examples using this technique demonstrate enhanced image resolution in various applications, including airborne SAR imaging, image processing for three dimensional interferometric ISAR imaging, and foliage-penetration ultrawideband SAR image processing. PMID- 15825488 TI - A comparison of rapid bovine spongiform encephalopathy testing methods on autolyzed bovine brain tissue. AB - In 1999, the European Union (EU) approved 3 rapid methods for the testing of bovine brain samples for the presence of bovine spongiform encephalopathy (BSE). The evaluation that led to the approval did not include an analysis of autolyzed material. Member states of the EU have active surveillance programs for BSE, which target fallen stock as well as other categories of cattle. Autolysis is a common feature of fallen stock samples because there can be a considerable delay between death and collection of samples. Therefore, it is important to know whether these tests perform optimally on autolyzed samples. The Veterinary Laboratories Agency (VLA) selected 250 positive fallen stock samples. These had been detected during routine testing using the Prionics-Check Western blot and confirmed as BSE cases by immunohistochemistry or electron microscopy. Samples were graded according to the degree of autolysis and then tested by the 3 methods: Prionics-Check Western blot, Platelia test, and Enfer test. All 3 methods correctly classified the samples as positive BSE cases, therefore alleviating doubt about their ability to do so. Subsequent EU validation exercises, such as those conducted in 2002--2003, have included the testing of autolyzed material. It is important that all new methods be evaluated on autolyzed tissue before approval for official use. PMID- 15825489 TI - Application of nox-restriction fragment length polymorphism for the differentiation of Brachyspira intestinal spirochetes isolated from pigs and poultry in Australia. AB - Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry. PMID- 15825490 TI - Comparison of ear notch immunohistochemistry, ear notch antigen-capture ELISA, and buffy coat virus isolation for detection of calves persistently infected with bovine viral diarrhea virus. AB - Two techniques performed on skin biopsy samples (ear notches), immunohistochemistry (IHC) and antigen-capture ELISA (AgELISA), were compared for detection of bovine viral diarrhea virus (BVDV) persistent infection (PI) in 559 Angus calves between the ages of 1 and 5 months. The calves also were tested for BVDV infection using virus isolation (VI) and reverse transcription (RT)-PCR on buffy coat samples and for antibodies to BVDV types la and 2 by serum neutralization (SN). Sixty-seven of 559 (12.0%) calves tested positive at initial screening by IHC, AgELISA, or VI, and all 67 were kept for a minimum of 3 months and retested monthly by IHC, AgELISA, VI, RT-PCR, and SN. Of the calves positive at initial screening, 59/67 (88.1%) were determined PI and 8/67 (11.9%) were determined acutely infected. Both IHC and AgELISA detected 100% of PI calves; however, IHC and AgELISA also detected 6 and 8 acutely infected calves, respectively, at initial screening. Furthermore, IHC and AgELISA continued to detect 3 and 4 acutely infected calves, respectively, 3 months after initial screening. Three acutely infected calves had IHC staining indistinguishable from PI calves at initial screening. Both IHC and AgELISA are accurate at detecting BVDV-infected calves, but veterinarians and producers should be advised that both tests detect some calves acutely infected with BVDV in addition to PI animals. Repeat testing using VI or RT-PCR on buffy coat samples should be performed at 30 days after initial screening to conclusively discriminate between acute and PI. PMID- 15825491 TI - Evaluation of a PCR to detect Salmonella in fecal samples of horses admitted to a veterinary teaching hospital. AB - The diagnostic accuracy of a PCR used to identify horses shedding Salmonella spp. in their feces during hospitalization was estimated, relative to bacterial culture of serially collected fecal samples, using longitudinal data. Five or more fecal samples were collected from each of 116 horses admitted as inpatients, for reasons other than gastrointestinal disease, between July 26, 2001 and October 25, 2002. All 873 fecal samples collected were tested with a PCR based on oligonucleotide primers defining a highly conserved segment of the histidine transport operon gene of Salmonella typhimurium, and each sample was cultured for Salmonella spp. One or more samples from 87 (75%) horses were PCR positive, and Salmonella was cultured from 1 or more samples from 11 (9.5%) horses. All culture positive horses had at least 1 PCR-positive result, whereas only 29 (28%) culture negative horses were PCR negative on all fecal samples tested. The PCR was most specific, relative to bacterial culture of serially collected fecal samples, when used to test samples from Quarterhorse or breeds other than Thoroughbred or Standardbred, or from clinical (vs. healthy, accompanying horses) cases. Overall, the PCR had the greatest agreement (70%), compared with bacterial culture of serially collected fecal samples, using a cutoff of 2 or more positive PCR test results to define a Salmonella-positive horse. The reasons why some fecal samples, from which Salmonella organisms cannot be isolated, are PCR positive need to be determined before the PCR can be incorporated into Salmonella surveillance programs for hospitalized equine populations. PMID- 15825492 TI - High-throughput real-time RT-PCR assay to detect the exotic Newcastle Disease Virus during the California 2002--2003 outbreak. AB - During the 2002--2003 Exotic Newcastle Disease (END) outbreak in Southern California, a high-throughput real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) system was developed to respond to the large diagnostic and surveillance sample workload. A 96-well RNA extraction method, using magnetic bead technology, combined with a 96-well RRT-PCR assay, allowed 1 technician to process and test more than 400 samples per day. A 3-technician team could complete testing on approximately 1,900 samples per day. The diagnostic sensitivity of the high-throughput RRT-PCR assay was 0.9967 (95% CI 0.9937 0.9997) based on 926 virus isolation confirmed positive samples. Diagnostic specificity using an initial 434 virus isolation confirmed negative samples was 100%. A diagnostic specificity of 0.9999 (95% CI 0.9999, >0.9999) was subsequently calculated on the basis of 2 false-positive results among 65,343 surveillance samples collected after the final END-positive case was confirmed in May 2003. Assay performance over 500 replicates, including reproducibility of the combined extraction and RRT-PCR amplification steps yielded a standard deviation of 0.70 RRT-PCR cycle thresholds (Ct) and a standard deviation of 0.59 Ct for the RRT-PCR steps alone. The high-throughput RRT-PCR developed for END contributed significantly to the 2002--2003 END control effort, reducing the predicted timeline for eradication from 3 years to just 11 months, primarily because of the large number of samples that could be rapidly tested. The 96-well approach described for high-throughput END RRT-PCR could be adapted to other rapid, high volume testing needs, as required for potential foreign animal disease responses or intensive surveillance efforts. PMID- 15825493 TI - Clinical and virological findings in pups naturally infected by canine parvovirus type 2 Glu-426 mutant. AB - An outbreak of canine parvovirus type 2 infection caused by the Glu-426 mutant in 2 litters of pups is reported. The infected pups (n = 6) were monitored daily for evidence of clinical signs and hematological changes and for the evaluation of viral shedding in the feces. The disease induced by the Glu-426 mutant was mild in all the infected pups. Vomiting and hemorrhagic diarrhea were not observed; however, the pups developed mucoid diarrhea (3.5 median days), depression (1.5 median days), and relative leukopenia and lymphopenia (2.5 median days). Fever and loss of appetite were observed only in 2 pups. Virus was detected in the feces for 4.5, 6.5, and 46 median days by hemagglutination, virus isolation on cell cultures, and real-time polymerase chain reaction (PCR), respectively. By real-time PCR, the highest viral DNA titers were detected in the feces of both litters at day 10, reaching median values of more than 10(10) DNA copies/mg of feces. PMID- 15825494 TI - C-reactive protein measurement in canine saliva. AB - An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments. PMID- 15825496 TI - Combined use of the ASK and SHK-1 cell lines to enhance the detection of infectious salmon anemia virus. AB - Infectious salmon anemia (ISA) is a severe disease primarily affecting commercially farmed Atlantic salmon (Salmo salar) in seawater. The disease has been reported in portions of Canada, the United Kingdom, the Faroe Islands, and the United States. Infectious salmon anemia virus (ISAV), the causative agent of ISA, has also been isolated from several asymptomatic marine and salmonid fish species. Diagnostic assays for the detection of ISAV include virus isolation in cell culture, a reverse transcriptase-PCR, an enzyme-linked immunosorbent assay, and an indirect fluorescent antibody test. Virus isolation is considered the gold standard, and 5 salmonid cell lines are known to support growth of ISAV. In this study, the relative performance of the salmon head kidney 1 (SHK-1), Atlantic salmon kidney (ASK), and CHSE-214 cell lines in detecting ISAV was evaluated using samples from both experimentally and naturally infected Atlantic salmon. Interlaboratory comparisons were conducted using a quality control-quality assurance ring test. Both the ASK and SHK-1 cell lines performed well in detecting ISAV, although the SHK-1 line was more variable in its sensitivity to infection and somewhat slower in the appearance of cytopathic effect. Relative to the SHK-1 and ASK lines, the CHSE-214 cell line performed poorly. Although the ASK line appeared to represent a good alternative to the more commonly used SHK-1 line, use of a single cell line for diagnostic assays may increase the potential for false-negative results. Thus, the SHK-1 and ASK cell lines can be used in combination to provide enhanced ability to detect ISAV. PMID- 15825495 TI - Immunohistochemical and ultrastructural detection of intestinal spirochetes in Thoroughbred horses. AB - Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen-containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen containing spirochetes were found in 7 of 12 horses (58.3%) and were more frequent in the cecum than in other parts of the bowel. It was not clear whether the infection was clinically related to diarrhea or dysentery, but histopathology revealed a close association between the bacterial infection and epithelial hyperplasia. Crypt epithelium consisted mainly of goblet cells and showed frequent mitosis throughout its length. Inflammatory cells and congestion were also present. There were numerous spirochetes in the crypts, and some invaded the cecal and colonic epithelia and underlying lamina propria. Ultrastructurally, the spirochetes were divided into 4 types. Three types were identified in degenerative epithelial cells or intracellularly. Brachyspira antigen-containing intestinal spirochetes invading the mucosa were capable of causing epithelial hyperplasia in the cecum and colon in the horses. The findings in this study will increase awareness of the importance of intestinal spirochetosis and may also be helpful for diagnosis and treatment of this condition. PMID- 15825497 TI - Comparative analysis of expert and machine-learning methods for classification of body cavity effusions in companion animals. AB - A rule-based expert system using CLIPS programming language was created to classify body cavity effusions as transudates, modified transudates, exudates, chylous, and hemorrhagic effusions. The diagnostic accuracy of the rule-based system was compared with that produced by 2 machine-learning methods: Rosetta, a rough sets algorithm and RIPPER, a rule-induction method. Results of 508 body cavity fluid analyses (canine, feline, equine) obtained from the University of California-Davis Veterinary Medical Teaching Hospital computerized patient database were used to test CLIPS and to test and train RIPPER and Rosetta. The CLIPS system, using 17 rules, achieved an accuracy of 93.5% compared with pathologist consensus diagnoses. Rosetta accurately classified 91% of effusions by using 5,479 rules. RIPPER achieved the greatest accuracy (95.5%) using only 10 rules. When the original rules of the CLIPS application were replaced with those of RIPPER, the accuracy rates were identical. These results suggest that both rule-based expert systems and machine-learning methods hold promise for the preliminary classification of body fluids in the clinical laboratory. PMID- 15825498 TI - Simultaneous detection of North American and European porcine reproductive and respiratory syndrome virus using real-time quantitative reverse transcriptase PCR. AB - Porcine reproductive and respiratory syndrome (PRRS) is 1 of the most economically important diseases of swine. Detection of the etiologic agent, PRRS virus (PRRSV), represents a diagnostic challenge due to the heterogeneity of field isolates as well as the propensity for swine to develop persistent infection in which virus is difficult to detect. Recently European (EU) lineage PRRSV isolates, which are genetically divergent from North American (NA) isolates, have been introduced into NA swine further complicating efforts to diagnose this disease. In this study, real-time (TaqMan) reverse transcriptase (RT)-PCR assays were developed for multiplex detection, differentiation, and quantification of NA and EU PRRSV field isolates. Oligonucleotide primers and dual-labeled probes were selected from conserved regions of open-reading frame 7 and the 3'-untranslated region. The real-time RT-PCR assays described for the NA or EU genotype of PRRSV detected viral RNA from 83/83 strains (74 NA; 9 EU) previously isolated by cell culture between 1992 and 2003. The analytical sensitivity of both assays was consistently found to be less than a single TCID50, which corresponded to 5-10 RNA molecules, and was not significantly reduced when the reactions were performed in a multiplex format. When performing multiplex reactions, sensitive detection was possible even when 1 viral RNA concentration was up to 5,000-fold higher than the second. The diagnostic sensitivity and specificity of the multiplex reaction was found to be at a minimum equivalent to that of both nested RT-PCR and virus isolation. PMID- 15825499 TI - Malignant catarrhal fever-like disease in sheep after intranasal inoculation with ovine herpesvirus-2. AB - A malignant catarrhal fever (MCF)-like disease was induced experimentally in 3 sheep after aerosol inoculation with ovine herpesvirus-2 (OvHV-2). Each of 3 OvHV 2-negative sheep was nebulized with 2 ml of nasal secretions containing approximately 3.07 X 10(9) OvHV-2 DNA copies from a sheep experiencing an intensive viral-shedding episode. Ovine herpesvirus-2 DNA became detectable by polymerase chain reaction in the peripheral blood leukocytes of all 3 sheep within 3 days, and all 3 seroconverted between 6 and 8 days postinfection (PI). The sheep developed clinical signs, with copious mucopurulent nasal discharge and fever around 14 days PI. One of the 3 clinically affected sheep was euthanized at 18 days PI. Major lesions at necropsy were multifocal linear erosions and ulcers in mucosa of the cheeks, tongue, pharynx, and proximal esophagus and mild disseminated pneumonia. Microscopically, there was extensive moderate superficial histiocytic-lymphocytic rhinitis with epithelial dissociation and degeneration. Moderate multifocal histiocytic bronchointerstitial pneumonia was associated with loss of terminal bronchiolar epithelium. Lymphocytic vasculitis was present only in the lung. The remaining 2 sheep recovered clinically, approximately 25 days PI. The study revealed that clinical signs and lesions resembling MCF can develop when uninfected sheep are exposed to a high dose of aerosolized OvHV-2. PMID- 15825500 TI - First outbreak of respiratory disease associated with swine influenza H1N2 virus in pigs in Korea. AB - This report describes the first diagnosis of swine influenza H1N2 virus infection in growing pigs in Korea by virus isolation, reverse transcription-polymerase chain reaction (RT-PCR), histopathology, and in situ hybridization. The subtype of swine influenza virus isolates was determined to be H1N2 by RT-PCR. The most consistent and predominant histological feature was bronchointerstitial pneumonia. Lung tissues from these pigs were hybridized with the nonradioactive digoxigenin-labeled complementary DNA (cDNA) probe from H1 HA and N2 NA genes but not from H3 HA and N1 NA genes. A strong hybridization signal was seen in bronchial- and bronchiolar-lining epithelial cells and in alveolar and interstitial macrophages. PMID- 15825501 TI - Validation of a human immunoturbidimetric assay to measure canine albumin in urine and cerebrospinal fluid. AB - The aim of this study was to validate an automated immunoturbidimetric assay used to quantify human albumin in urine and to accurately measure canine albumin concentrations in both urine and cerebrospinal fluid. The partial homology existing between human and canine albumin limited the accuracy of the human assays in measuring canine albumin without method modifications. Thus, the assay was modified by calibrating the analyzer with calibrators made in the laboratory containing known concentrations of canine albumin. To prepare the set of calibrators, the albumin concentration of pooled sera of healthy dogs was assessed in 5 replicates using the BromocresolGreen assay. Pooled samples were aliquoted and serially diluted to obtain the expected concentrations of albumin (0.5, 1, 5, 13, and 30 mg/dl) for establishing the canine calibration curve. Thereafter, the performance was assessed by analyzing canine urine and CSF The modified assay accurately quantified canine albumin in both specimens, as indicated by the following. Intra- and interassay variability was 0.92% and 2.74%, respectively; recovery was 99.66% and 99.07% in urine and 105.02% in CSF No interference was detected when hemolysate and glucose were added to urine. The test was linear within the verified range (0-225 mg/dl). These results demonstrate that the modified human albumin immunoturbidimetric assay can be a useful tool in the veterinary diagnostic laboratory. It is accurate and tends itself to automatization on chemistry analyzers. PMID- 15825502 TI - Detection of Echinococcus multilocularis infection in a colony of cynomolgus monkeys (Macaca fascicularis) using serology and ultrasonography. AB - Five animals in a colony of cynomolgus monkeys (Macaca fascicularis) died or were euthanatized because of alveolar echinococcosis, during a period of 5 years. The remainder of the colony was screened for possible infection with Echinococcus multilocularis, using serology and ultrasonography. A total of 46 animals out of a group of 55 were examined. The presence of anti-Em2 antibodies analyzed with enzyme-linked immunosorbent assay was demonstrated in 3 monkeys. In 2 of these 3 monkeys, multilocular structures compatible with metacestodal cysts in the liver were identified, using ultrasonography. The presence of alveolar echinococcosis was subsequently confirmed at postmortem examination in 1 animal. The other animals are still alive. Two other monkeys were negative in the serological examination but had cystic structures in the liver, which were identified as bile duct cysts at postmortem examination in 1 animal. The other monkey is still alive. These findings suggest that serology for antibodies against the Em2 antigen may represent a useful method in identifying animals that might be infected with E. multilocularis and are therefore at risk of developing fatal alveolar echinococcosis. PMID- 15825503 TI - Enterotoxemia associated with beta2 toxin-producing Clostridium perfringens type A in two Asiatic black bears (Selenarctos thibetanus). AB - Beta2 (beta2) toxin-producing Clostridium perfringens type A strains were found to be associated with necrotic and hemorrhagic intestinal lesions in 2 Asiatic black bears (Selenarctos thibetanus) that died suddenly. Ten isolates were obtained from the liver, lungs, heart, and small and large intestine of the animals and were examined by multiplex polymerase chain reaction for the genes encoding the 4 lethal toxins (alpha, beta, epsilon, and iota) for classification into toxin types as well as for the genes encoding enterotoxin and the novel beta2-toxin for subclassification. In addition, the cpb2 sequence of the 10 isolates was different from the published sequence of cpb2 of pig type C isolate CWC245, whereas it was highly similar to the cpb2 sequence of the C. perfringens type A strain 13. This finding suggests the existence of 2 cpb2 subtypes. This is the first report of enterotoxemia associated with the presence of C. perfringens producing beta2-toxin in the tissues and intestinal content of Asiatic black bears. PMID- 15825504 TI - Detection of bovine leukemia virus by in situ polymerase chain reaction in tissues from a heifer diagnosed with sporadic thymic lymphosarcoma. AB - An 18-month-old bovine heifer was presented for clinical evaluation after a sudden onset of ventral edema. Clinical and pathological evaluations were consistent with thymic lymphosarcoma, a sporadic form of lymphosarcoma in cattle, which is not generally considered to be associated with bovine leukemia virus (BLV). This heifer was seropositive for BLV at 6 and 18 months of age. Tissues obtained at necropsy were evaluated using in situ polymerase chain reaction. The BLV proviral DNA was detected in lymphocytes of the thymus as well as in epithelial cells of the liver and kidney. This report presents evidence that thymic lymphosarcomas can be associated with BLV infection and that BLV may have a broader cellular tropism than was supposed previously. PMID- 15825505 TI - Neuronal ceroid-lipofuscinosis in a Holstein steer. AB - A young, partially blind Holstein steer was affected by mild cerebral atrophy. Formalin-fixed cerebral gray matter was diffusely yellow brown. Microscopically, there were eosinophilic, autofluorescent granules primarily in the cytoplasm of cerebral neurons. There was also extensive retinal atrophy with complete loss of the rod and cone layers. Ultrastructural examination of affected cerebral neurons revealed a mixture of granular osmiophilic and lamellar patterns in the cytoplasmic storage bodies. This suggests the existence of neuronal ceroid lipofuscinosis in the Holstein breed. PMID- 15825506 TI - Environmental air sampling to detect exotic Newcastle disease virus in two California commercial poultry flocks. AB - The 2002--2003 Exotic Newcastle Disease (END) outbreak in Southern California poultry provided an opportunity to evaluate environmental air sampling as an efficient and cost-effective means of sampling flocks for detection of a circulating virus. Exotic Newcastle Disease virus was detected by real-time reverse transcriptase PCR from air samples collected using a wetted-wall cyclone style air sampler placed within 2 m of birds in 2 commercial flocks suspected of being naturally exposed to END virus during the outbreak. Exotic Newcastle Disease virus was detected after 2 hours of air sampling the poultry-house environments of the 2 naturally infected flocks. PMID- 15825508 TI - Health literacy. PMID- 15825507 TI - Fatal mycobacteriosis with hepatosplenomegaly in a young dog due to Mycobacterium avium. AB - Cases of disseminated Mycobacterium avium infections in dogs are rare because it appears that the species is innately resistant to infection. A 2-year-old, castrated, 5 kg Shih Tzu-Poodle-cross developed anemia, abdominal pain, lethargy, and splenomegaly. Histological examination of surgically removed spleen indicated marked granulomatous splenitis with myriad intracytoplasmic acid-fast bacterial rods. Ultrastructural examination revealed the presence of 3-4-microm-long mycobacteria in phagolysosomes of epithelioid macrophages. Tissue extract of lightly fixed spleen was positive for M. avium 16S ribosomal RNA and negative for M. tuberculosis complex IS6110 DNA by polymerase chain reaction testing. Anemia was associated with the presence of mycobacteria-infected macrophages in bone marrow. The animal's condition deteriorated, and euthanasia was performed after a clinical course of 2 months. The principal morphological findings at necropsy were severe diffuse granulomatous hepatitis, enteric lymphadenomegaly, and segmental granulomatous enteritis with intralesional mycobacteria present. Mycobacterium avium was cultured from enteric lymph nodes sampled at necropsy. The source of infection was not established but was presumed to be environmental with an enteric portal of entry. PMID- 15825509 TI - Support Bill C-22. PMID- 15825510 TI - Method of euthanasia in food animals must be practical and safe. PMID- 15825511 TI - Dispensing barbiturates to swine producers. PMID- 15825512 TI - Logical to vaccinate rather than risk infection. PMID- 15825513 TI - An ethicist's commentary on a feedlot using a drug in an unapproved way. PMID- 15825514 TI - Laboratory confirmed positive cases of equine West Nile virus in Alberta in 2003. PMID- 15825515 TI - Evaluation of gastrointestinal activity patterns in healthy horses using B mode and Doppler ultrasonography. AB - Healthy adult horses were examined by using transabdominal ultrasonography to quantitatively and qualitatively evaluate activity of the jejunum, cecum, and colon with B mode and Doppler techniques. Doppler ultrasound was used to assess jejunal peristaltic activity. Examinations were performed on multiple occasions under imposed colic evaluation conditions, including fasting, nasogastric intubation, and xylazine sedation. In fasted horses, jejunal visibility was increased and jejunal, cecal, and colonic activity was decreased. The stomach was displaced ventrally and was visualized ventral to the costochondral junction. Xylazine sedation in fed horses had minimal effects; however, in fasted horses, xylazine significantly decreased jejunal and cecal activity. Nasogastric intubation in fasted horses had no observable effects on activity, but moved the stomach dorsally. B mode and Doppler jejunal activity were strongly correlated. Prior feeding and sedation status need to be considered when interpreting the results of equine abdominal ultrasound examinations. Doppler techniques may be useful for assessing jejunal activity. PMID- 15825516 TI - Antimicrobial resistance of selected Salmonella isolates from food animals and food in Alberta. AB - Salmonella isolates (n = 209) obtained from food animals and foods in Alberta during 1996 through 1999 were tested for sensitivity to 17 antimicrobials. Of the 3553 antimicrobial susceptibility tests on Salmonella isolates, 11.8% were positive for resistance. These isolates were commonly resistant to tetracycline (35.4%), streptomycin (32.5%), sulfamethoxazole (28.7%), ticarcillin (27.3%), and ampicillin (26.8%). Resistance to at least 1 antimicrobial was observed in 112 isolates (53.6%). Salmonella Typhimurium, S. Typhimurium var. Copenhagen, and S. Heidelberg were the most common serovars among isolates resistant to individual antimicrobials and multiple antimicrobials. The most common profile of multiple antimicrobial resistance was that which included resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline, and ticarcillin. The proportions of isolates that were resistant to antimicrobials were greater among bovine isolates of Salmonella than among poultry isolates, and this difference was greater among isolates from veterinary diagnostic sources than among those from monitoring sources. PMID- 15825517 TI - Effects of a single dose of ketamine on duodenal motility activity in pigs. AB - In order to investigate the effects of a single dose of ketamine on duodenal motility, the present study focused on the electric impedance technique. Five pigs (32 to 40 kg, CVC group) were instrumented with a central venous catheter 1 d before measurements. The next day, general anesthesia was started and maintained via central venous catheter by propofol and fentanyl. In contrast, the pigs of the KETA group (n = 5) received ketamine intramuscularly prior to the induction of anesthesia by the injection of propofol-fentanyl via an ear vein. An intraluminal impedance catheter was manually introduced into the proximal duodenum. Measurements were recorded for 4 h. The KETA group showed a median duration of phase II that was shortened by 35%, while phase I was prolonged by 73% (P < 0.05). In conclusion, when gastrointestinal motility has to be investigated, the effects of a single dose of ketamine, even for premedication, should be taken into consideration. PMID- 15825518 TI - Efficacy of a saponin-adjuvanted inactivated respiratory syncytial virus vaccine in calves. AB - The objective of this study was to determine whether a commercially available, saponin-adjuvanted, inactivated bovine respiratory syncytial virus (BRSV) vaccine would protect calves from experimental infection with virulent BRSV. This was a randomized controlled trial comprising 14, 8- to 9-week-old calves seronegative for BRSV Group 1 calves (n = 8) were not vaccinated and group 2 calves (n = 6) were vaccinated on days 0 and 19 with an inactivated BRSV vaccine. All calves were challenged with virulent BRSV on day 46. Clinical signs, arterial PO2, and immune responses were monitored after challenge. Calves were euthanatized on day 54 (8 d after challenge) and lungs were examined for lesions. Vaccination elicited increases in BRSV-specific immunoglobulin (Ig) G and virus neutralizing antibody titers. Challenge with BRSV resulted in severe respiratory tract disease and extensive pulmonary lesions in control calves, but no signs of clinical disease and minimal or no pulmonary lesions in vaccinated calves. Arterial blood oxygen values on day 53 (7 d after challenge) in control calves were significantly lower than those in vaccinated calves, which remained within normal limits. Control calves shed BRSV for several days after challenge, whereas BRSV was not detected on deep nasal swabs from vaccinated calves. In summary, the results indicated that this inactivated BRSV vaccine provided clinical protection from experimental infection with virulent virus 27 d after vaccination and significantly decreased the prevalence and severity of pulmonary lesions. Efficacy was similar to that reported for other commercial inactivated and modified-live BRSV vaccines. PMID- 15825519 TI - Primary splenic mesenchymal chondrosarcoma in a dog. AB - A large splenic mass was found during exploratory celiotomy in a 12-year-old Labrador retriever. Histologic examination of the mass revealed a mesenchymal chondrosarcoma. The dog was euthanatized 7 wk postoperatively, due to suspected tumor recurrence. Extraskeletal mesenchymal chondrosarcomas are rare in dogs and are associated with a guarded prognosis. PMID- 15825520 TI - Choanal lymphosarcoma in a 7-year-old golden retriever: diagnosis and treatment. AB - A 7-year-old, spayed female, golden retriever with a 1-year history of progressive stertor and reverse sneezing was referred for further diagnostics. Endoscopy and computed tomography of the head revealed a bilobed mass in the choanae. Cytologic and histopathologic examinations of the mass were consistent with lymphosarcoma. Treatment consisted of chemotherapy. PMID- 15825521 TI - Orthopedic odds and ends. PMID- 15825522 TI - The art of chronic medical problems. PMID- 15825523 TI - Two cases of multicentric lymphosarcoma in swine: gross and histopathologic findings. PMID- 15825524 TI - Superficial pyoderma, secondary to the hairless nature of the breed. PMID- 15825525 TI - The Pan-American Association of Ophthalmology: an opportunity for the Americas. PMID- 15825526 TI - RESIO revisited: visual function assessment and cataract surgery in British Columbia. AB - BACKGROUND: Cataract surgery in a population with mild levels of functional impairment was studied in the Regional Evaluation of Surgical Indications and Outcomes (RESIO) project. The RESIO investigators assessed the overall visual function assessment (VFA) score before and after surgery. In this study we reanalysed available RESIO data to better understand the findings for individual VFA items in this patient population. METHODS: We calculated effect sizes for longitudinal changes in each VFA item and for changes in overall VFA scores using the Mann-Whitney statistic. We considered effect sizes less than 0.3 as small, between 0.3 and 0.8 as moderate, and more than 0.8 as large. We compared differences in the individual VFA item scores before and after cataract surgery using a paired t test and Wilcoxon's signed-rank test. RESULTS: Of the 32 surgeons originally involved in the RESIO project, 18 (60%) agreed to have the data for their patients reanalysed. We found pre- and postoperative VFA data for 709 cataract operations (mean age of patients 73.4 years [standard deviation 9.4 years]; 60.7% women). Of the 47 people with a perfect preoperative VFA score, 28 (59.6%) had a perfect score postoperatively, 13 (27.6%) had a score of 90 or better, 5 (10.6%) had a score of 87 to 90, and 1 (2.1%) had a score of 67. Among the 200 people with a preoperative VFA score of 90 to 99, postoperatively the VFA score improved for 146 (73.0%), remained the same for 43 (21.5%) and worsened for 11 (5.5%). The mean scores for all individual VFA items improved postoperatively. The largest improvements were noted for "driving during the day" (effect size 1.1), "self-care activities" (0.8), "driving during the night" (0.5), "doing fine handiwork" (0.5), "reading newspaper or book" (0.4) and "participating in religious activities" (0.4). INTERPRETATION: The original conclusion from the RESIO project seems correct: based on VFA, the current threshold for cataract surgery in British Columbia is very low. Individual VFA items, particularly reading and night driving, are most strongly and consistently influenced by cataract surgery, regardless of overall VFA score. In the absence of visual acuity data, it is not possible to relate VFA to cataract surgical decision making in British Columbia. PMID- 15825527 TI - Consent to cataract surgery performed by residents. AB - BACKGROUND: Surgical teaching seems to be in conflict with the contract between surgeon and patient. We carried out a study to determine the prevalence of consistent disclosure to patients that a resident will perform part or all of their cataract surgery procedure. A second objective was to investigate the effect of such disclosure on patients' willingness to undergo the procedure. METHODS: We sent a survey to all 20 ophthalmologists working in our university affiliated hospitals, inquiring about their practice of disclosure to patients regarding residents' involvement in surgery. Staff physicians were also asked to record their patients' consent to an operation performed partly or entirely by a trainee while under supervision. RESULTS: Of the 20 surveys sent, only 5 (25%) were returned. Those who declined to participate in the study mentioned several reasons, including that such disclosure might increase a patient's anxiety level, that they might lose potential patients as patients might be reluctant to have trainees perform their surgery, and lack of time to talk to patients about these issues. Of the five ophthalmologists who completed the survey, four were part time affiliated staff and one was a geographic full-time physician working in our institution. Four of the five ophthalmologists said that they do not consistently disclose residents' involvement to their patients. Of the 49 patients enrolled, only 8 (16%) agreed to undergo the procedure after being informed that a trainee would be actively involved. INTERPRETATION: It is crucial to inform patients that residents may be involved in their surgery in order to avoid possible litigation. However, our results suggest that such disclosure may have a negative effect on surgical education because it could limit the number of cases available to trainees. PMID- 15825528 TI - Anophthalmia and microphthalmia in the Alberta Congenital Anomalies Surveillance System. AB - BACKGROUND: A higher than expected rate of anophthalmia/microphthalmia (A/M) for 1999 was noted in both the Alberta Congenital Anomalies Surveillance System (ACASS) and the Canadian Congenital Anomalies Surveillance System (CCASS). Since this increase was at variance with the previous 19 years, we performed a review to determine whether the increase was true and, if so, the possible explanation. METHODS: We reviewed the records of the cases of A/M in the ACASS together with the accompanying attachments (e.g., consultant, autopsy and chromosome reports) for 1991-2001. In addition, we contacted all 91 registered ophthalmologists in Alberta. Letters were also written to the Edmonton and Calgary offices of the Canadian National Institute for the Blind (CNIB). RESULTS: Sixty cases of A/M were ascertained over the study period. Of the 88 active ophthalmologists in the province, 21 (24%) replied, but no new cases were ascertained from this source. No replies were received from the CNIB. We constructed five categories of clinical phenotypes for the 60 cases: 20 had a chromosomal etiology, 13 had a recognized syndrome or association, 16 had extraocular malformations, 5 had other eye anomalies, and 6 had A/M only. Pregnancy terminations were not included. The higher rate in 1999 was mainly due to cases with a chromosomal etiology or a recognized syndrome or association. There was no indication that a teratogen was causing a cluster of A/M cases, as our annual rates were comparable to those for other jurisdictions not only in Canada but also in other countries. INTERPRETATION: Our review confirmed that the rate of A/M in Alberta in 1999 was high but that the increase was mainly due to five cases of trisomy 13 together with one case associated with a syndrome (Meckel-Gruber). Our findings provide reassurance that there was no environmental cause of clustering of anophthalmia or microphthalmia. This review demonstrates the importance of ongoing population based surveillance in providing baseline birth prevalence rates for evaluating trends and clusters. PMID- 15825529 TI - Effects of a 0.9% sodium chloride ophthalmic solution on the ocular surface of symptomatic contact lens wearers. AB - BACKGROUND: Ocular surface-related discomfort is the main reason for stopping contact lens wear. We carried out a study to evaluate the efficacy of preservative-free artificial tears containing 0.9% sodium chloride on ocular surface signs and symptoms in contact lens wearers experiencing discomfort and its possible influence on the duration of contact lens wear. METHODS: We studied 49 contact lens wearers experiencing discomfort who had normal results of slit lamp biomicroscopy, a fluorescein tear film break-up time (BUT) of 10 seconds or more, and wetting greater than 5 mm in 5 minutes on the Schirmer 1 test with and without anesthesia. Twenty-nine subjects (16 men and 13 women with a mean age of 32.5 years [standard deviation (SD) 8.7 years]) received one instillation of the 0.9% sodium chloride solution four times daily in the lower conjunctival fornix for 21 days. Twenty subjects (12 men and 8 women with a mean age of 35.1 [SD 6.2] years) received no drops and served as a control group. The overall comfort and duration of contact lens wear, results of tear film analysis and adverse events were recorded on days 7 and 21. Patients rated their symptoms (while not receiving any medications or hydrating solutions) on a 100-mm visual analogue scale with "Excellent (lenses not felt)" at the left and "Very uncomfortable (lenses cause irritation or discomfort)" at the right. Measurement of corrected visual acuity, slit-lamp examination, determination of the tear film BUT, the Schirmer 1 test with and without anesthesia, and assessment of the colour and surface of the lens were performed at baseline and at day 21. We analysed the data for the more uncomfortable eye or, if the eyes were equally uncomfortable, the right eye. RESULTS: Significant lessening of ocular discomfort was observed in the treatment group during the study: the mean rating on the visual analogue scale at baseline was 60.2 mm (SD 12.7 mm), compared with 35.8 mm (SD 18.0 mm) at day 21 (p < 0.001, Student's t test). The duration of contact lens wear was significantly longer at day 21 than at baseline (7.0 [SD 2.6] hours vs. 6.4 [SD 2.6] hours, p < 0.05, Student's t test), and the proportion of subjects with conjunctival hyperemia was significantly lower at day 21 (48.3% vs. 82.8%, p < 0.05, chi2 test). No statistically significant changes were observed in tear film BUT, results of the Schirmer 1 test, corneal punctate staining by fluorescein or results of tear film analysis. The treatment was well tolerated by all patients. No significant differences in any of the variables studied were observed in the control group. INTERPRETATION: Treatment with a preservative-free 0.9% sodium chloride ophthalmic solution reduced ocular surface discomfort and extended the duration of contact lens wear without interfering with the tear film or contact lens materials. Long-term studies are needed to confirm the role of this solution in reducing discomfort experienced by contact lens wearers. PMID- 15825530 TI - Effect of the menstrual cycle on standard achromatic and blue-on-yellow visual field analysis of women with migraine. AB - BACKGROUND: It has been postulated that migraine and glaucoma may have common vascular causative factors. Significant sex-based differences in the incidence of many important ocular conditions raise the possibility that estrogens may have direct effects on the eye. We performed a study to determine the effect of the menstrual cycle on standard achromatic automated perimetry (SAP) and short wavelength automated perimetry (SWAP) (blue-on-yellow perimetry) of women with migraine. METHODS: Both eyes of 73 normally menstruating women (31 subjects with migraine and 42 healthy control subjects) were included in the study. Subjects underwent a complete ocular examination including SAP and SWAP in both the follicular phase (12th to 13th day of the cycle) and the luteal phase (1 to 2 days before the onset of bleeding) of two consecutive menstrual cycles.We performed visual field analysis using the Humphrey Field Analyzer II with the full-threshold central 30-2 program. Mean sensitivity was calculated for the superior temporal, inferior temporal, superior nasal and inferior nasal regions separately. RESULTS: Thirteen subjects were lost to follow-up (5 in the migraine group and 8 in the control group), leaving 26 subjects and 34 subjects respectively. There was no significant difference in mean age between the two groups (33.9 years [standard deviation (SD) 3.4 years] vs. 35.1 years [SD 3.3 years]). The mean duration of migraine was 7.6 (SD 3.1) years (range 3-14 years). In both groups, serum estradiol levels were significantly lower (p = 0.001) and serum progesterone levels were significantly higher (p < 0.001) in the luteal phase than in the follicular phase. In the control group, the mean sensitivity values with SWAP were significantly lower in the luteal phase than in the follicular phase (p = 0.04). A similar decrease was observed for the subjects with migraine with both SAP and SWAP (p = 0.01). There was no difference in regional mean sensitivity between the two phases with either perimetric test in the control group. For the subjects with migraine, there was no difference in regional mean sensitivity between the two phases with SAP. However, with SWAP, the mean sensitivity for the nasal visual field locations was significantly lower in the luteal phase than in the follicular phase (p = 0.01). INTERPRETATION: Our study provides further evidence of an effect of sex hormones on the visual field of women with migraine. In addition to assessment of intraocular pressure, menstrual cycle phases should be considered in women with migraine at risk for glaucomatous optic neuropathy. PMID- 15825531 TI - Evaluation of a new preoperative ophthalmic solution. AB - BACKGROUND: Maximum mydriasis and corneal clarity during intraocular surgery are important to ensure operational safety. However, repeated instillation of mydriatic and anti-inflammatory ophthalmic solutions during surgery may affect compliance and may damage the corneal epithelium. We developed an ophthalmic solution containing tropicamide, phenylephrine hydrochloride and diclofenac sodium, and compared the properties and effect on corneal epithelial barrier function and on mydriasis of the solution and of its individual components. METHODS: We developed the ophthalmic solution by mixing 10 mL of 0.5% tropicamide/0.5% phenylephrine, 10 mL of 5% phenylephrine and 10 mL of 0.1% diclofenac (TPD). We determined the stability of the chemicals in solution using high-performance liquid chromatography immediately, 1 week and 1 month after mixing. Corneal epithelial barrier function was assessed before and after instillation of the solutions in 26 eyes of 17 patients (5 with proliferative diabetic retinopathy or diabetic maculopathy, 8 with cataract and 4 with no eye disease). The fluorescent intensity was measured 10 times at the central cornea 30 minutes after instillation of 3 mL of 0.5% fluorescein solution and the average value calculated. Finally, the pupil diameter of 50 eyes of 38 patients undergoing cataract surgery was measured before and immediately after the operation. RESULTS: No remarkable changes in the pH or pharmacologic activity of the TPD solution were observed at any time after mixing. In the patients with diabetic retinopathy or cataract, the increase in mean fluorescent intensity was significantly greater with the individual solutions than with the TPD solution (p < 0.01); there was no significant difference in mean fluorescent intensity in the subjects with no eye disease. No statistically significant difference in pupil diameter was observed between the eyes that received the TPD solution and those that received the individual solutions. INTERPRETATION: TPD ophthalmic solution is simple to prepare and use. The TPD solution had a similar effect on mydriasis as the three individual solutions but was less destructive to the corneal epithelium. PMID- 15825532 TI - Complications of intravitreal injection of triamcinolone acetonide. AB - BACKGROUND: Intravitreal injection of triamcinolone acetonide appears to be a promising treatment for a variety of proliferative, edematous, neovascular and inflammatory ocular disorders. Reported complications include intraocular pressure (IOP) elevation, cataract formation, retinal detachment, vitreous hemorrhage and endophthalmitis. The purpose of this investigation was to report the complications of intravitreal triamcinolone injection that may be attributable to the injection procedure or to the corticosteroid suspension. METHODS: A total of 212 eyes of 180 patients who underwent intravitreal triamcinolone acetonide injection for various indications were enrolled. All patients received 8 mg/0.2 mL of triamcinolone. A total of 270 injections were performed by the same surgeon under topical anesthesia. The patients were followed for a mean of 9.2 months. Complications related to the injection procedure and to the corticosteroid were recorded. RESULTS: The most common complication encountered during follow-up was transient elevation of the IOP above 21 mm Hg (44 eyes [20.8%]). The average IOP rose by 28.5%, 38.2%, 16.7% and 4.2% from baseline at 1, 3, 6 and 9 months respectively. The mean IOP values at 1, 3 and 6 months were statistically significantly higher than the mean preinjection value (p < 0.001). Fourteen eyes (6.6%) had cataract progression and underwent cataract surgery with intraocular lens implantation. Endophthalmitis developed in one eye (0.5%); the patient underwent vitrectomy with silicone oil injection. Pseudoendophthalmitis occurred in one eye (0.5%), and pseudohypopyon was observed in two eyes (0.9%). INTERPRETATION: Intravitreal triamcinolone injection was effective in a variety of ocular disorders. Patients should be monitored closely given the potential for complications of the injection procedure or the corticosteroid suspension. PMID- 15825533 TI - Subconjunctival spread of ciliary body melanoma after glaucoma filtering surgery: a clinicopathological case report. PMID- 15825534 TI - Metastatic choroidal melanoma presenting as a solitary pulmonary nodule. PMID- 15825535 TI - Bilateral Candida chorioretinitis: involvement of the second eye after 3 years. PMID- 15825536 TI - Acinetobacter corneal graft ulcer and endophthalmitis: report of two cases. PMID- 15825537 TI - Alacrima presenting in the third decade. PMID- 15825538 TI - Intractable secondary glaucoma from hyphema following YAG iridotomy. PMID- 15825539 TI - Combined retinal artery and vein occlusion in a patient with systemic lupus erythematosus and antiphospholipid syndrome. PMID- 15825540 TI - Sweet's syndrome presenting as acute episcleritis. PMID- 15825541 TI - Review of awakening agents. AB - Brain injuries are a serious burden of illness to Canada and the US. Advances in managing head trauma have allowed more patients to emerge from decreased levels of consciousness and helped them cope with neurocognitive, neurobehavioural, and neuropsychiatric deficits. In this article, we review the current (1986-2002) evidence surrounding the pharmacological management of arousal states and the aforementioned neurological sequelae of head injury in either acute or chronic conditions. This article will review the evidence for the use of psychostimulants (methylphenidate), antidepressants (amitriptyline, selective serotonin reuptake inhibitors, and buproprion), Parkinson's medications (amantadine, bromocriptine, carbidopa/levodopa), anticonvulsants (valproic acid), modafinil (Provigil), lactate, hyperbaric oxygen chamber, electroconvulsive therapy, and transmagnetic stimulation, in patients following a head injury. The review did not include all anticonvulsants, neuroleptics, beta-blockers, benzodiazepines, azospirones or cognitive enhancers. Unfortunately, the quality of the evidence is generally poor, and sometimes conflicting, which in turn results in indecisive guidelines for treating patients. Accepting the inherent flaws in the evidence we feel that this paper may serve as a stepping-stone for future researchers to improve data gathering that targets neurocognitive, neurobehavioural and neuropsychiatric symptoms following a head injury. PMID- 15825542 TI - Progress in clinical neurosciences: Advances in the management of low-grade gliomas. AB - The management of low-grade gliomas represents one of the most challenging and controversial areas in neuro-oncology. Many aspects of the treatment of low-grade gliomas are debated, including the optimal timing of surgery and radiotherapy, the benefit of extensive surgery, and the impact of these variables on the natural history of these indolent and generally incurable tumours. The recently published results of several large multicentre trials addressing the timing and dose of radiotherapy have provided solid evidence for delayed and reduced dose irradiation. These studies have also confirmed prognostic variables that can be used to guide management of individual patients. Among these variables is the observation that tumours with oligodendroglial features have a better natural history and response profile. The recognition that as many as two thirds of low grade gliomas have oligodendroglial features, advances in molecular diagnostics making accurate pathologic diagnosis of oligodendroglial tumours possible, and the established chemosensitivity of malignant oligodendrogliomas, have raised new issues surrounding the potential value of chemotherapy for low-grade gliomas. This review will be restricted to low-grade diffuse astrocytomas, oligodendrogliomas, and low-grade mixed oligoastrocytomas in adults, and provide evidence-based guidelines for the management of these tumours, including the emerging role of chemotherapy as initial treatment. PMID- 15825543 TI - Canadian Association of Neuroscience review: Cellular and synaptic insights into physiological and pathological pain. EJLB-CIHR Michael Smith Chair in Neurosciences and Mental Health lecture. AB - Neurons and synapses in the central nervous system are plastic, undergoing long term changes throughout life. Studies of molecular and cellular mechanisms of such changes not only provide important insight into how we learn and store new knowledge in our brains, but they also reveal the mechanisms of pathological changes that occur following injury. The author proposes that during induction, neuronal mechanisms underlying physiological functions, such as learning and memory, may share some common signaling molecules with abnormal or injury-related changes in the brain. Distinct synaptic and neuronal network mechanisms are involved in pathological pain as compared to cognitive learning and memory. Nociceptive information is transmitted and regulated at different levels of the brain, from the spinal cord to the forebrain. Furthermore, N-methyl-D-aspartate receptor-dependent and calcium-calmodulin activated adenylyl cyclases (AC1 and AC8) in the anterior cingulate cortex play important roles in the induction and expression of persistent inflammatory and neuropathic pain. Neuronal activity in the anterior cingulate cortex can also influence nociceptive transmission in the dorsal horn of the spinal cord by activating the endogenous facilitatory system. Our results provide important synaptic and molecular insights into physiological responses to injury. PMID- 15825544 TI - Incidence and prevalence of multiple sclerosis in Newfoundland and Labrador. AB - BACKGROUND: The incidence and prevalence of multiple sclerosis (MS) in Newfoundland and Labrador (NL) had been reported in 1984 and was considered to be relatively low at that time. This study revisits the incidence and prevalence of MS in NL for the year 2001. METHODS: Case searches through patient files of neurologists in NL were conducted. A complete list of patients billed for MS in NL between 1996 and 2003 was obtained and all cases were confirmed via chart review. RESULTS: There were 493 living MS patients yielding a prevalence of 94.4/100,000 which is significantly higher than previously reported. Of the living patients, 330 had relapsing remitting (RRMS), 94 had secondary progressive, 66 had primary progressive (PPMS) and three had unspecified MS. The total female to male ratio was 2.7:1. There was no difference between the female to male ratios for RRMS vs PPMS. Patients with PPMS had a later onset compared to RRMS (p<0.00001). Yearly incidences were relatively constant from 1994 to 2001 (5.6/100,000). Significant delays between first symptoms and final diagnosis were common and the delay time has not changed over the past 15 years. A prevalence of 88.9/100,000 was estimated from survival and incidence trends and was not significantly different than the measured prevalence (p=0.38). CONCLUSIONS: The increase in incidence and prevalence are accounted for through both better access to diagnostic facilities and more practicing neurologists. The revised prevalence and incidence are more in keeping with recently reported values throughout Canada. PMID- 15825545 TI - Multiple sclerosis in Newfoundland and Labrador--a model for disease prevalence. AB - BACKGROUND: Newfoundland and Labrador, Canada, have been almost exclusively populated by immigrants from southwest England and southeast Ireland. The province's population grew largely by natural increase from 20,000 people in 1835 to half a million at present. Very little interregional migration occurred within the province. This uniquely-populated region and its subsequent founder effect provide the basis to develop models of disease prevalence. OBJECTIVES: To develop a model for the regional prevalence of multiple sclerosis (MS), accounting for settlement patterns and geographic location (latitude). METHODS: All living MS patients with confirmed addresses (438 patients) in the province were mailed a survey requesting their place of birth. Regional prevalences were calculated from a 75% rate of return of the survey. Theoretical regional prevalences were proportionally calculated from the source prevalences of southwest England, southeast Ireland, Scotland and the Channel Islands based on settlement patterns. These theoretical regional prevalences were corrected for geographical variations of latitude based on observations in the United Kingdom. Theoretical and actual regional prevalences were compared. RESULTS: When actual regional prevalences were compared with theoretical prevalences, very little variation was noted, especially after correcting for variation in latitude. CONCLUSION: A regional variation in MS prevalence is noted in the island portion of Newfoundland and Labrador. This regional variation can be modeled by using both migration patterns and latitudinal position. This model demonstrates that the prevalence of MS is influenced by both genetic and environmental contributions. PMID- 15825546 TI - Indicators of functional status for primary malignant brain tumour patients. AB - BACKGROUND: We compared the functional status and survival time of patients with malignant gliomas. METHODS: This retrospective review included 143 patients diagnosed with malignant gliomas. Patients were grouped according to histopathological diagnosis. To measure functional status, patients were assigned a Karnofksy performance status (KPS) score at the time of presentation and at one, three, six, nine, 12 months and yearly intervals thereafter. Data were analyzed using descriptive methods as well as Kruskal-Wallis tests, Chi-square tests, Log-Rank tests and Cox's proportional hazards modeling. RESULTS: Eighty four patients were male. The median age of patients was 63 years. One hundred and seven patients had a histopathological diagnosis of glioblastoma multiforme, 23 of anaplastic astrocytoma and 13 of anaplastic oligodendroglioma. Twenty-nine patients received aggressive multimodal treatment, 83 received intermediate treatment and the remaining 31 patients received conservative therapy. Significant treatment complications occurred in 33% of patients including four post-operative deaths. The anaplastic oligodendroglioma group had lower mortality and maintained better KPS scores over time, as did patients receiving full treatment. The most significant prognostic factors for functional status included age, pretreatment KPS, and type of treatment received. The most significant factors associated with time until death included age, severity of comorbidities, pretreatment KPS, presence of confusion, histopathological diagnosis and type of treatment received. CONCLUSION: In patients with malignant gliomas, younger age, better functional status at presentation and aggressive multimodal treatment were associated with improved longer-term functional status and survival. Confirmation of the effect of multimodal treatment on patient functional status would require a randomised controlled clinical trial. PMID- 15825547 TI - Anterograde ventricular irrigation during fourth ventricle region surgery. AB - OBJECTIVE: To reduce postoperative hydrocephalus following surgery in the region of the fourth ventricle. METHOD/TECHNIQUE: We describe the use of a conventionally placed lateral ventricular catheter to facilitate anterograde ventricular irrigation during surgery in the region of the fourth ventricle. This technique offers a safe alternative to more conventional approaches to fourth ventricular irrigation and obviates the need for placement of a third or fourth ventricular catheter and/or serial lumbar punctures by thoroughly flushing blood and tissue debris from the operative field at the time of surgery. RESULTS: Preliminary results spanning a five-year period show a lower incidence in the need for early ventriculo-peritoneal shunts (5%) and delayed ventriculo peritoneal shunts (10%). CONCLUSION: This technique represents a safe method of reducing hydrocephalus following operative procedures in the region of the fourth ventricle. PMID- 15825548 TI - The BRAINS study: safety, tolerability, and dose-finding of repinotan in acute stroke. AB - BACKGROUND AND PURPOSE: Repinotan is a potent, serotonin (5-HT1A) full receptor agonist that interferes with ischemia-mediated neuronal cell death in animal models. This double-blind, placebo-controlled phase II study examined the safety, tolerability, and dose of repinotan in patients with acute ischemic stroke. METHODS: Patients with acute hemispheric ischemia and a National Institutes of Health Stroke Scale of 4 to 25 were randomized to placebo or repinotan 0.5, 1.25, or 2.5 mg/day (d) given by continuous intravenous infusion for 72 hours. Treatment was started within six hours of symptom onset. Evaluations were performed at four weeks and three months. RESULTS: Among 240 patients in the safety analysis, repinotan was well-tolerated, with adverse events appearing more frequently in the highest dose group (2.5 mg/d). The most common adverse event was headache (21.3% to 35% with repinotan and 24.1% with placebo). Most (75%) adverse events were of mild or moderate severity. The most common severe adverse events were neurological worsening, cerebral hemorrhage, and brain edema. The number of deaths and serious adverse events were similar among placebo and repinotan groups. Compared to the placebo group, the proportion of patients with good outcomes at three months was greatest in the group receiving repinotan 1.25 mg/d, although the difference was not statistically significant. CONCLUSIONS: This study indicates that the incidence of adverse events was comparable with all doses of repinotan and placebo, and no safety issues were observed. A trend toward better tolerability with evidence of efficacy was observed with the repinotan 1.25 mg/d dose. PMID- 15825549 TI - Neurofilament M gene in a French-Canadian population with Parkinson's disease. AB - BACKGROUND: Recently, a single base pair substitution (G1747A) mutation of the neurofilament M (NF-M) gene was reported in a French-Canadian patient with early onset Parkinson's disease (PD). Three unaffected siblings were found to be heterozygotes for the NF-M Gly336Ser mutation but, to date, no other affected PD individuals have been found with a similar mutation. No other individuals with Parkinson's disease and of similar ethnic background have been screened for this mutation. METHODS: We screened 102 French-Canadian patients with definite PD and 45 French-Canadian controls for this substitution in the NF-M gene using a PCR restriction enzyme digestion method. RESULTS: None of the patients or controls carried this mutation. CONCLUSION: Our results would indicate that this mutation is not common even in a PD population of similar ethnic background and suggest this change represents a rare variant. However, these results do not exclude the possibility that other mutations in this gene could be present. PMID- 15825550 TI - Microstent-assisted coiling for wide-necked intracranial aneurysms. AB - OBJECTIVE: To describe the results, technical feasibility, efficacy and challenges encountered in our preliminary experience using a self-expandable microstent, optimized for intracranial use, as an adjunct in the endovascular treatment of wide-necked aneurysms. METHODS: Only broad-necked aneurysms (dome-to neck ratio < or = 2, or an isolated neck size > 4.5 mm) were treated with Neuroform microstent from July 2003 to May 2004. The techniques used for stent deployment were either parallel or sequential. Angiographic results were recorded immediately for all patients and classified as Class 1 (complete occlusion), Class 2 (neck remnant) or Class 3 (sac remnant) by three interventional neuroradiologists not involved in the procedure. Follow-up angiography at six months was obtained for one case. Modified Rankin Score scale was assessed for all patients. RESULTS: Seventeen intracranial aneurysms in a total of 18 patients were treated (mean age, 52.2 yr). Eight patients (44.4%) presented with acute subarachnoid hemorrhage. Eleven aneurysms (61.1%) were in the posterior circulation. Average dome size was 10.2 mm (range, 3.7-19.8 mm) and average neck size was 5.36 mm (range, 3.0-10.0 mm). Six out of seven aneurysms of the anterior circulation were approached with parallel technique. Eight aneurysms of the posterior circulation were approached with sequential technique. Average number of coils deployed was 9.64 (range, 4-23 coils). Eleven aneurysms (64.8%) resulted in Class 1 and/or Class 2. One technical failure was observed. Technical complications were recognized in four patients (23.5%), all of them with unruptured aneurysms in the anterior circulation. Two patients (11.7%) presented transient immediate clinical complications. One patient (5.8%) had minor permanent neurological complication. Neither major clinical complications nor death were encountered. Favorable clinical outcome (Modified Rankin Scale score 0 2) was observed in 88.2% of the patients (average follow-up time, 4.72 months). CONCLUSIONS: Absence of major permanent complications and satisfactory immediate obliteration degree in our preliminary experience indicates that microstent assisted coiling technique is useful for the minimally invasive treatment of broad-necked complex aneurysms that are not ideal for conventional endovascular treatment and are at a high risk for conventional surgical treatment. PMID- 15825551 TI - Electrogastrography in patients with Parkinson's disease. AB - BACKGROUND: Impaired gastrointestinal motility in Parkinson's disease may affect absorption of levodopa and contribute to the disabling response fluctuations (RF). In this study gastric myoelectric activity was recorded with electrogastrography in patients with PD and correlated with the duration, severity and the presence of RF. METHOD: Electrogastrography (EGG) was performed in 36 patients with PD of which 22 were men. The mean age was 67 years (48-81), mean duration of disease was 7.07 years (1-20), and mean duration of treatment with levodopa was 5.07 years (1-20). Gastric dysrhythmia was diagnosed when either preprandial or postprandial dysrhythmia for more than 30% of the recording period was detected. RESULTS: The EGG was abnormal in 24 of 36 patients. Significant association was found between preprandial dysrhythmia and duration of disease (P=0.002); duration of levodopa treatment (P=0.003), severity of 86RF (P=0.001), but not with age (P=0.076). Out of 18 patients with RF, 17 had at least one pattern of dysrhythmia. In 11 out of the 18 patients without RF, the EGG was normal while the remaining seven had at least one pattern of dysrhythmia. CONCLUSION: Abnormal EGG was quite common in this group of patients with PD, particularly in those with RF. The most common pattern of abnormality was preprandial dysrhythmia, which was positively associated with disease duration and length of levodopa treatment. Although frequently asymptomatic, preprandial dysrhythmia leading to impaired gastric emptying may contribute to irregular absorption of levodopa from the small intestine and contribute to disabling response fluctuations. PMID- 15825552 TI - Severe traumatic brain injury in a large Canadian health region. AB - BACKGROUND: Although severe traumatic brain injury (sTBI) is a devastating condition with tremendous public health implications, the epidemiology of this disease has not previously been described in Canada. We sought to define the incidence, risk factors and outcome of patients suffering sTBI in a large Canadian region. METHODS: A population-based surveillance cohort design was utilized to identify all Calgary Heath Region residents who were victims of trauma with an injury severity score > or = 12. Subsequent application of a specific sTBI case definition defined the final cohort. RESULTS: The annual incidence of sTBI was 11.4 per 100,000 population. The incidence of sTBI was significantly higher for males as compared to females [17.1 vs. 5.9 per 100,000; relative risk (RR) = 2.91, 95% confidence interval; 2.17, 3.94; p<0.0001]. There was a striking increase in the annual age specific population incidence of sTBI observed among those older than 74 years of age. The relative risk among the highest risk group of elderly (>85 years) males as compared to the lowest risk female group (50-64 years) was 19.78 (95% CI; 6.27, 62.3; p<0.0001). One hundred and eight patients died prior to hospital discharge for a mortality rate of 5.1 per 100,000 per year. CONCLUSIONS: Severe traumatic brain injury is common among residents of the Calgary Health Region and is associated with a high mortality rate. Males and the elderly are at the highest risk for acquiring sTBI and may represent target groups for preventive efforts. PMID- 15825553 TI - Clobazam as add-on therapy for temporal lobe epilepsy and hippocampal sclerosis. AB - BACKGROUND: Clobazam is a benzodiazepine with known antiepileptic action; however, it is not considered first line therapy in the treatment of epilepsy. The objective of this study was to evaluate the efficacy of clobazam as add-on therapy in adults with temporal lobe epilepsy associated with MRI evidence of hippocampal sclerosis (HS). METHOD: This is a retrospective study, conducted at our epilepsy clinic which evaluated clobazam as add-on therapy in patients with temporal lobe epilepsy and MRI signs of HS. Clobazam was prescribed based on the minimum effective dose up to the maximum tolerated dose. RESULTS: Seventy-eight patients met the inclusion criteria (51 women), ages ranging from 16 to 76 years old (mean=42.2). Dosage of clobazam ranged from 5 to 60 mg/day (mean=22.6 mg/day). Clobazam was used from one month to eight years (mean=29 months). Sixteen (20.5%) patients were seizure-free, 20 (25.5%) had more than 75% improvement in seizure control, eight (10%) had more than 50% and 20 (26%) were non responders to clobazam. In 14 (18%) we could not determine seizure frequency during follow-up. The improvement in seizure control lasted for more than one year in 30 (68%) patients. CONCLUSION: Our data suggest that clobazam should be considered as add-on therapy in the treatment of patients with temporal lobe epilepsy associated with MRI signs of HS. PMID- 15825554 TI - A spontaneous recurrent seizure bioassay for anti-epileptogenic molecules. AB - Drug design in epilepsy is now tackling a new target--epileptogenesis. This is the process whereby a normal brain becomes susceptible to recurrent seizures. One of the stumbling blocks in the design and discovery of new chemical entities as antiepileptogenics is the implementation of an appropriate biological model. Current models, such as the maximal electroshock model, are models of seizures, not models of epileptogenesis. To develop such a model, we have extended and modified a chronic pilocarpine spontaneous recurrent seizure (SRS) model for the purposes of developing a bioassay with which to screen new compounds for putative antiepileptogenic bioactivity. PMID- 15825555 TI - A 58-year-old woman with progressive vertigo, deafness and weakness. PMID- 15825556 TI - Calcium pyrophosphate dihydrate deposition disease resulting in cervical myelopathy. PMID- 15825557 TI - An infant with central nervous system complications of disseminated tuberculosis. PMID- 15825558 TI - Cerebral small vessel disease in pseudoxanthoma elasticum: three cases. AB - BACKGROUND: Cerebral small vessel disease is rarely described in association with pseudoxanthoma elasticum (PXE), a hereditary connective tissue disorder with skin, eye and vascular manifestations. This autosomally inherited elastic tissue disease has been attributed to mutations in the ABCC6 gene located on chromosome 16p13.1. Different stroke mechanisms are suggested in PXE patients, arterial hypertension and accelerated atherosclerosis being the leading ones. CASE DESCRIPTIONS: Case 1: A 49-year-old man with history of mild hypertension presented with recurrent transient ischemic attacks. At the age of 42, evaluation for progressive visual loss and skin changes led to diagnosis of PXE. Brain magnetic resonance imaging (MRI) disclosed multiple lacunar infarctions and confluent periventricular white matter lesions (WML). Case 2: A 71-year-old woman with history of mild hypertension suffered right-sided stroke. Diagnosis of PXE was made at the age of 48 due to severe visual loss and skin changes. Brain MRI revealed multiple lacunar infarctions and subcortical ischemic leukoencephalopathy. Case 3: A 47-year-old woman with prominent skin changes and bilateral amblyopia developed right-sided weakness. Skin biopsy confirmed PXE. Several lacunar infarcts in deep white matter and pons were revealed on MRI. DISCUSSION: We present three patients with clinical and histopathological features of PXE who presented with multiple lacunar strokes, two with extensive confluent WML. These cases illustrate that PXE is a rare but significant risk factor for small vessel disease and stroke in patients of all age groups. Occlusive small vessel disease and subsequent lacunar infarcts and WML represent important PXE manifestations. PMID- 15825559 TI - Acute bilateral opercular strokes causing loss of emotional facial movements. AB - The classic anterior opercular syndrome of Foix-Chavany-Marie presents with loss of voluntary facial, pharyngeal, lingual, and mastication movements, with preservation of emotional and automatic movements. Most commonly, sequential strokes affecting bilateral opercula cause this syndrome. The inverse clinical presentation, with selective loss of emotional facial movements, has only rarely been reported, and is less well-localized. CASE REPORT: We report a case of selective loss of emotional facial movements which resulted from bilateral acute infarcts. No etiology was discovered, and the syndrome was reversible. DISCUSSION: The available literature, and findings in this case, suggest that voluntary and automatic facial movements have distinct pathways, and damage to the insula bilaterally may lead to the selective loss of emotional facial movements. The clinical presentation of this inverse automatic/voluntary dissocation needs to be recognized as a rare syndrome with bilateral localization, so that patients at higher risk of further stroke can quickly be identified. PMID- 15825560 TI - Sex steroid and growth factor profile of a meningioma associated with pregnancy. AB - BACKGROUND: Increased growth of meningiomas during pregnancy as well as postpartum clinical regression of symptoms have been reported but remain poorly understood. A better understanding of the factors that contribute to these observations, including potential factors associated with pregnancy, could enable design of more effective adjuvant therapies. METHODS: We describe the presentation of a meningioma during the immediate postpartum period. Serial imaging demonstrated subsequent rapid decrease in size of the tumour prior to any intervention. The lesion was resected, and the tissue was subjected to immunostaining for gene products associated with pregnancy, including estrogen receptor (ER), progesterone receptor (PR), platelet-derived growth factor receptor B (PDGFRB), fibroblastic growth factor receptor 2 (FGFR-2), epidermal growth factor receptor (EGFR) and human placental lactogen (hPL). RESULTS: The lesion proved to be an atypical fibroblastic meningioma grade II (WHO). Immunostaining demonstrated significant staining for PR, PDGFRB, and FGFR-2. No specific staining for ER, EGFR, or hPL was identified. CONCLUSION: Although clinical regression of meningioma following pregnancy is well-recognized, imaging data are much less abundant. This report provides clear clinical and imaging documentation of a meningioma associated with pregnancy. In addition, the growth factor profile of this tumour suggests the importance of PR, PDGFRB, and FGFR-2 as potential therapeutic targets. PMID- 15825561 TI - Vecuronium induced prolonged paralysis in two pediatric intensive care patients. PMID- 15825562 TI - Re: Exacerbation of pre-existing epilepsy by mild head injury. Tai PC, Gross DW. Can J Neurol Sci 2004; 31:394-397. PMID- 15825563 TI - Re: Comparison of monitoring techniques for intraoperative cerebral ischemia. DW Rowed, DA Houlden, LM Burkholder, AB Taylor. Can J Neurol Sci 2004;31(3):347-356. PMID- 15825564 TI - Flexible endoscopic evaluation of swallowing with sensory testing: patient characteristics and analysis of safety in 1,340 consecutive examinations. AB - Flexible endoscopic evaluation of swallowing with sensory testing (FEESST) is a comprehensive endoscopic assessment of the sensory and motor components of a swallow. Previous studies addressing patient safety issues with respect to FEESST included relatively small numbers of patients and paid almost no attention to patient characteristics. The purpose of this study was to determine the incidence of FEESST-related complications in the outpatient and inpatient settings and to analyze patient diagnoses that led to the performance of FEESST. We performed a prospective study of FEESST complications in 1,340 consecutive evaluations performed over a 4 1/2-year period. The primary outcome variables were incidence of epistaxis and airway compromise. The secondary outcome variable was underlying patient diagnoses. The incidence of epistaxis was 1 in 1,340 (0.07%). There were no instances of airway compromise. Stroke was the most common reason for the performance of FEESST (343; 25.6%), followed by cardiac-related dysphagia (298; 22.2%) following open heart surgery (169/298; 56.7%), heart attack, congestive heart failure, or new arrhythmia. The remaining causes were head and neck cancer (207; 15.4%), pulmonary disease (141; 10.5%), chronic neurologic disease (124; 9.3%), and acid reflux disease (80; 6.0%). We conclude that FEESST is a relatively safe procedure for the sensory and motor assessment of dysphagia in a cohort of patients with a wide variety of underlying diagnoses. The emergence of cardiac surgery as a common cause of dysphagia warrants further study. PMID- 15825565 TI - Meta-analysis of upper probe measurements in normal subjects and patients with laryngopharyngeal reflux. AB - We report a meta-analysis of a series of studies in which 24-hour ambulatory pH monitoring was performed in 1) normal subjects, 2) the normal control subjects in studies of laryngopharyngeal reflux (LPR), and 3) the patients with LPR in these controlled studies. The statistical analysis utilized the fixed-effects model by Mantel-Haenszel and the random-effects mixed model. There were 16 studies from the past 12 years that fulfilled the inclusion criteria. They involved 793 subjects (264 normal and 529 with LPR). The numbers of positive pharyngeal reflux events for normal subjects and for patients with LPR differed with a p value of <.0001. There was also a significant difference in the mean percentage of acid exposure times between normal subjects and patients with LPR (p = .003). We conclude that the upper probe gives accurate and consistent information in normal subjects and patients with LPR. The numbers of reflux events and acid exposure times are most important in distinguishing normal subjects from patients with LPR. The technology and methodology of probe testing is quite reliable and is consistent on a worldwide basis. PMID- 15825566 TI - Histologic characterization of rat vocal fold scarring. AB - This study aimed to clarify the characteristics of rat vocal fold scarring by examining the alteration of key components in the extracellular matrix: hyaluronic acid, collagen, and fibronectin. Under monitoring with a 1.9-mm diameter telescope, unilateral vocal fold stripping was performed, and larynges were harvested at 2, 4, 8, and 12 weeks after operation. The vocal folds were histologically analyzed with Alcian blue stain, trichrome stain, and immunofluorescence of collagen type I, collagen type III, and fibronectin. The scarred vocal folds showed less hyaluronic acid and more collagen types I and III than did the controls at all time points. Type III was stable for 12 weeks, while type I declined until 8 weeks and thereafter remained unchanged. Fibronectin increased for 4 weeks and then decreased; it was close to the control level at 8 and 12 weeks. These results suggest that the tissue remodeling process in scarred vocal folds slows down around 2 months after wounding. PMID- 15825567 TI - Bile-induced laryngitis: is there a basis in evidence? AB - Most agree that bile reflux occurs with regularity in an otherwise healthy population and that biliary and acid reflux may play a synergistic role in damaging esophageal mucosa. But to what extent is laryngeal mucosa at risk? We constructed a saline-controlled rat model (n = 40) in which active component solutions of bile--taurocholic acid and chenodeoxycholic acid--were applied to intact laryngeal mucosa at various pH levels. Histologic sampling of the laryngeal mucosa allowed inflammation scores to be generated by a pathologist blinded to the solutions used. Both taurocholic acid at acid pH and chenodeoxycholic acid at basic pH preferentially induced statistically greater inflammation scores than did the saline control, approaching or exceeding inflammation scores attributed to hydrochloric acid at pH 1.2. These observations may clarify reasons for failure to uniformly control laryngeal injury by adequate suppression of gastric acid alone and may further justify alternative methods of laryngeal protection in patients refractory to adequate acid control. PMID- 15825568 TI - Pulsed dye laser treatment of laryngeal granulomas. AB - Laryngeal granulomas are effectively treated with antireflux therapy and speech therapy. Failure to respond leads to treatment with Botox or surgical excision. We report on the use of the pulsed dye laser for treating chronic granulomas that do not respond to standard therapy. We performed a retrospective review from September 2002 to September 2003. Patients identified with chronic granulomas that were not responding to standard therapy were treated in our office with the pulsed dye laser. Ten patients were identified; the mean age was 58 years. Two patients underwent more than one pulsed dye laser treatment. Five of the 10 had resolution of their lesions, and 3 had a partial response. Two were unchanged. The average follow-up was 6 months, and there were no complications. We conclude that in-office use of the pulsed dye laser is a relatively safe and effective method for treating laryngeal granulomas that do not respond to antireflux therapy and speech therapy. PMID- 15825569 TI - Unilateral vocal cord paralysis causes contralateral false-positive positron emission tomography scans of the larynx. AB - Positron emission tomography (PET) is used in the management of head and neck cancers. It identifies tissue with increased metabolic activity and is not specific for malignancy. A false-positive PET scan of the larynx is associated with vocal cord paralysis. We reviewed PET scan reports of patients with lung cancer from 1998 to 2001 to identify patients with increased 18-fluoro-2 deoxyglucose uptake in the larynx without a known history of head and neck cancer and then correlated this increased uptake with laryngoscopic findings. There were 17 patients who had a positive PET finding in the larynx. Fifteen of those had a false-positive PET scan in the larynx. All had contralateral vocal cord paralysis. Two patients were noted to have head and neck cancer. We conclude that vocal cord paralysis can cause a false-positive PET scan on the contralateral side of the larynx due to overactivity of laryngeal muscles that compensate for the paralyzed cord. PMID- 15825570 TI - Direct revascularization is superior for rat parathyroid allotransplantation with FK506. AB - Parathyroid gland allotransplantation has been a challenging goal for decades. Our objective was to optimize a parathyroid allotransplantation model for analysis of short-term or low-dose immunosuppressive regimens. Rats that had undergone parathyroidectomy received parathyroid allografts either by direct microvascular anastomoses as part of a composite laryngotracheal graft or by direct implantation into hind limb muscle. All rats were maintained on daily low dose FK506 (tacrolimus). Intact serum parathyroid hormone (PTH) levels for both groups were recorded over a period of at least 45 days and compared with a repeated-measures mixed model. We found that microvascular anastomosis was superior to implantation for parathyroid gland survival, as all revascularized grafts immediately produced normal levels of PTH, whereas implanted grafts had a significantly slower recovery of function (p < .001). Four of the 11 implanted grafts (36%) never produced detectable PTH levels. Using this model, we are developing innovative strategies that will lead to successful parathyroid allotransplantation without the need for chronic immunosuppression. PMID- 15825571 TI - Sudden sensorineural hearing loss in lupus erythematosus associated with antiphospholipid syndrome: case report and review. AB - Systemic lupus erythematosus is a disease of unknown cause that manifests with tissue and cellular alterations due to the deposition of antibodies and pathogenic immune complexes. The disease can be associated with anticardiolipin antibody syndrome, a disorder of recurrent vascular thrombosis and thrombocytopenia associated with a persistent anticardiolipin test positivity. In this report, we describe the case of a young woman affected by lupus erythematosus and positive for anticardiolipin antibodies who was brought to our observation for a sudden sensorineural hearing loss in the right ear. The patient was successfully treated with osmotic therapy and, 13 months after the initial symptom, is now in good clinical condition with no recurrence of the hearing loss and normal bilateral hearing. Together with a critical review of the pertinent literature, this rare clinical case led us to formulate some original comments. PMID- 15825572 TI - Purulent chondritis of the laryngeal framework cartilages. AB - This manuscript reports on our experience with purulent chondritis of the laryngeal cartilages (PCLC), an entity that has not yet been described. Three patients had a diagnosis of PCLC. The probable causes were relapsing polychondritis, a previous prolonged intubation, and an idiopathic cause. The patients suffered from hoarseness and inspiratory stridor for 1 to 3 months before diagnosis. None complained of pain in the neck. Laryngoscopy showed supraglottic edema. A computed tomography scan revealed abscess formation between the intact inner and outer perichondria of the thyroid cartilage. The treatment included rigid endoscopy, external incision and drainage, and prolonged medical therapy. The culture results were Staphylococcus aureus in the first 2 cases and Aspergillus fumigatus in the third. The second patient (in whom the cricoid cartilage was also affected) required emergency tracheotomy. The other 2 patients did not require airway intervention. The rarity of PCLC and the relatively mild symptoms require a high index of suspicion for its diagnosis. PMID- 15825573 TI - Optimal stimulus intensity and reliability of air stimulation technique for elicitation of laryngo-upper esophageal sphincter contractile reflex. AB - To determine the optimal air stimulus intensity and duration for elicitation of the laryngo-upper esophageal sphincter (UES) contractile reflex, we studied 37 healthy volunteers 20 to 81 years of age. A sleeve device monitored the UES pressure. For laryngeal stimulation, we used an air stimulator unit (Pentax AP 4000) that incorporated a nasolaryngeal endoscope. The arytenoids and interarytenoid areas were stimulated at least three times by three different stimuli: 6-mm Hg air pulse with 50-ms duration, 10-mm Hg air pulse with 50-ms duration, and 6-mm Hg air pulse with 2-second duration. Of 1,165 air stimulations, 1,041 resulted in mucosal deflections. Of these, 451 resulted in an abrupt increase in UES pressure. The response/deflection ratio for 6-mm Hg stimulation with 2-second duration was significantly higher than those for air pulses with 50-ms duration (p < .001). We conclude that although the laryngo-UES contractile reflex can be elicited by an air pulse with 50-ms duration, this ultrashort stimulation is not reliable. Using longer-duration pulses (at least 2 seconds) improves the reliability of elicitation of the laryngo-UES contractile reflex. PMID- 15825574 TI - Morphological and radiologic evaluation of the human nasopalatine duct. AB - In several mammals, a direct connection between the nasal cavity and the oral cavity is a common finding. The structure is named the nasopalatine duct (NPD). It has been hypothesized to be functional in terms of transportation of odorants from the oral cavity to the nasal cavity. In humans, the NPD exists during embryological development. The connection between the nasopalatine infundibulum and the incisive fossa is typically closed at the time of birth. We present the case of a 24-year-old man who presented with a persistent NPD. By means of a thin, soft tube, it was possible to probe the NPD. Magnetic resonance imaging revealed a bony gap between the hard palate and the alveolar process of the maxillary bone with a length of 16 mm and a width of 6 mm. In light of the literature of the past 500 years, this represents a rare finding of a persistent NPD. PMID- 15825575 TI - Taste disorders after tonsillectomy: case report and literature review. AB - The authors describe a case of dysgeusia that occurred during the recovery period after a tonsillectomy. The cause was thought to be a lesion to the lingual branch of the glossopharyngeal nerve because of the location of the symptoms at the posterior region of the buccal cavity and because of the raised electrogustometric thresholds in the posterior region of one half of the tongue. Clarification of this type of case was made by a review of the literature from 1966 to June 2004, carried out with the aid of Medline. From a medicolegal standpoint, it is important to inform the patient of the risk of dysgeusia after tonsillectomy, especially if that patient has a profession in which taste plays an essential role. PMID- 15825576 TI - Use of fibrin glue as a hemostatic in endoscopic sinus surgery. AB - Endoscopic sinus surgery (ESS), especially when combined with turbinectomy and/or with submucous resection of the septum, may involve postoperative bleeding that might end with nasal packing. Nasal packing causes pain, rhinorrhea, and inconvenience and may not stop the postoperative bleeding. The aim of our study was to compare the hemostatic properties of the second-generation surgical sealant Quixil (Crosseal) with those of nasal packing in ESS. We performed a prospective randomized trial in 64 consecutive patients who underwent ESS and presented excessive intraoperative and/or postoperative bleeding. They were allocated by the sealed-envelope method into two groups. A routine ESS procedure was ended with Merocel nasal packing in group 1, and with aerosol application of Quixil sealant at the operative site in group 2. The hemostatic effects were evaluated objectively in the clinic by anterior rhinoscopy and endoscopy and assessed subjectively by the patients at follow-up visits. In group 1, various types of postoperative bleeding occurred in 25% of patients. In group 2 there was no postoperative bleeding, except for 1 case of late hemorrhage (3.12%). Drainage and ventilation of the paranasal sinuses were not impaired. There were no allergic reactions to the glue. We conclude that aerosol application of fibrin glue can be readily performed in ESS, requires no special treatment (antibiotics), and appears to have an adequate hemostatic effect. The use of this second-generation glue in ESS appears to stop nasal bleeding well and to be relatively safe and convenient. PMID- 15825577 TI - Ossicular discontinuity and exostoses in Proteus syndrome: a case report. AB - Proteus syndrome (PS) is a rare hamartomatous disorder characterized by mosaic overgrowth of multiple tissues that manifests early in life and is progressive. The presence of unilateral external auditory canal exostoses in a patient who is not a swimmer or surfer is suggestive of PS. However, hearing loss is not a typical feature. Here, we describe exostoses and ossicular discontinuity with conductive hearing loss in a patient with PS. The treatment consisted of canalplasty and ossicular chain reconstruction. A postoperative reduction was demonstrated in the patient's air-bone gap, from 21 dB to 13 dB for the pure tone average (four frequencies) and from 41 dB to 15 dB in the high-frequency range (6,000 to 8,000 Hz). Causes of ossicular discontinuity are discussed. Routine annual audiometric and otolaryngological evaluation should be considered in all patients with temporal bone involvement of PS. PMID- 15825578 TI - Response of visual loss in allergic fungal sinusitis to oral corticosteroids. AB - Visual loss associated with allergic fungal sinusitis is most often treated with surgery followed by oral corticosteroids. A case is presented in which, because of substantial medical comorbidities, surgery could not be initially performed and the visual loss was corrected with prednisone alone. This case serves to reinforce the central role of corticosteroids in treatment of this enigmatic condition. PMID- 15825579 TI - Anatomy of the cricothyroid articulation: differences between men and women. AB - A greater difficulty in exposing the arytenoid cartilage during rotation surgeries has been observed for men. The objective of the present study was to describe the position of the cricothyroid articulation and the distance between the right and left articulations, and to compare these findings between genders. The following measurements were obtained for 16 cadavers (9 men and 7 women): angulation of the cricothyroid articulation in the cricoid ring, dimensions of the thyroid articular surface of the cricoid cartilage, and distance between the cricothyroid and cricoarytenoid articulations. The cricothyroid articulation angle was narrower in men than in women (p = .04). The major diameter of the articular facet of the thyroid cartilage was wider in men (p = .001). The longer lamina of the thyroid cartilage, as well as the more posterior position of the cricothyroid articulation, in men might explain the greater difficulty in exposing the arytenoid cartilage during laryngeal framework surgeries observed for this group of patients. PMID- 15825581 TI - Trauma and dissociation in conversion disorder and chronic pelvic pain. AB - OBJECTIVE: The purpose of this study was to: a) assess the link between sexual and/or physical abuse and dissociation in conversion disorder and chronic pelvic pain patients; and b) assess whether this effect is independent of level of general psychopathology. METHOD: This report examines data from four separate samples. Fifty-two patients with chronic pelvic pain, 61 patients with non epileptic seizures, and two samples of patients (102 and 54) with predominantly motor or sensory types of conversion disorder were studied. RESULTS: Using point biserial correlations no compelling evidence for a consistent and positive association of sexual and/or physical abuse with dissociation was found. After statistically controlling for level of psychopathology using multiple regression analyses, in most of the cases the association of abuse with dissociation was no longer statistically significant. Only physical abuse predicted level of somatoform dissociation over and above level of psychopathology. CONCLUSION: In future clinical studies of dissociation in patients with conversion disorder and chronic pelvic pain more complex models may be needed with less exclusive reliance on historical antecedents such as childhood abuse and more emphasis on recent potentially traumatizing experiences or co-morbid psychiatric disorders. PMID- 15825580 TI - Does antidepressant adherence have an effect on glycemic control among diabetic antidepressant users? AB - OBJECTIVE: To examine the relationship between adherence to antidepressant medications and HbA1c levels among patients with diabetes in a managed care setting. METHOD: The analysis included measures of HbA1c levels before, during, and after initial antidepressant use among 568 patients with diabetes enrolled in the Harvard Pilgrim Health Care insurance plan from 1991-1995. Adherence was defined as four refills in a six-month period after the first antidepressant prescription. General linear models using SAS PROC MIXED were used to estimate the effects of covariates including antidepressant adherence on HbA1c levels over time, comparing patients who were adherent to antidepressant medications to those patients who were non-adherent to antidepressant medications. RESULTS: Adherence to antidepressant treatment was not significantly associated with HbA1c levels among diabetic patients who are antidepressant users. Younger age, use of insulin and oral medications, and female gender were all significantly associated with HbA1c levels over time. CONCLUSIONS: Although we did not observe any association between level of adherence to antidepressant therapy among diabetic patients and levels of glucose control, our results confirm previously established associations between patient characteristics and glycemic control. Further research is needed to disentangle the complex relationship among antidepressant treatment adherence and diabetes outcomes. PMID- 15825582 TI - Information needs of recently diagnosed cancer patients in Brazil. AB - OBJECTIVE: The communication and disclosure of medical information to cancer patients has been an important issue in oncology, reflecting changes in the physicians' approach when dealing with the patients' ethical right to receive information. The aim of this study was to assess the general and specific information desired by cancer patients regarding their health conditions and to evaluate minor psychiatric symptoms. METHOD: Two-hundred and ninety-eight patients were examined at an oncological cancer center in Sao Paulo, Brazil, using a questionnaire designed to investigate patients' preferences about cancer information and evaluate minor psychiatric symptoms. RESULT: Ninety-five percent of the patients were interested in obtaining information on their health condition, with regard to whether they had cancer (95%), the chances of recovery (89%), and the side effects during treatment (94%). Younger patients tended to show more interest in obtaining information than the elderly. The Self-Reporting Questionnaire classified 25.8% of the sample as a probable psychiatric case, but no correlation was found between Self-Reporting Questionnaire scores and patients' desires for medical information. CONCLUSION: The majority of patients wished to be informed about their illness condition, and no correlation was found between psychiatric morbidity and desire for information. PMID- 15825583 TI - Detection and management of depressive disorders in primary care in Spain. AB - OBJECTIVE: Many patients with depression who attend primary care are not detected and are unaware of the benefits of therapeutic intervention. Our aim is to evaluate the recognition and management of depressed patients presenting in primary care in Spain. METHODS: This was a two-phase cross-sectional study in primary care centers in Tarragona (Spain). In the first phase, we screened 906 consecutive patients using the Zung's Self-Rating Depression Scale. In the second phase, all 209 patients whose results were positive and 97 patients whose results were negative (1/7 at random) were given the Structured Clinical Interview for DSM-IV Disorders, a series of questionnaires, and the judgment of the physician about the presence of depression was determined. In the 120 patients with major depression and/or dysthymia, predictors of detection and treatment of depression were determined. RESULTS: Seventy-two percent of depressed patients were detected. The detection was associated with educational level, severity of the depression, level of impairment, and the complaint of explicit psychological symptoms. Thirty-four percent were receiving treatment with antidepressants. Antidepressant treatment was associated with marital status, severity of and impairment from the depression, frequency of visits to the family physician, and the patients' complaint of psychological symptoms. CONCLUSION: Many depressed patients are not detected, but these are often the patients who present with the mildest forms of depression and for whom the benefits of detection are far from clear. Although treatment was associated with the severity of depression, most depressed patients, at any level of seriousness, do not receive appropriate treatment. PMID- 15825584 TI - Screening for mental illness in primary care clinics. AB - OBJECTIVES: To assess the prevalence of mental disorders in inner city outpatient clinics and to improve the diagnosis of mental illness in primary care. METHODS: The Problem Oriented Patient Report (POPR), a patient self-report checklist, was administered to 362 outpatients at two inner-city Buffalo primary care clinics. Patients' completed POPR checklists were evaluated to identify those with potential mental illness diagnoses and were available for the physicians' review during the patients' visits. After the visit, clinical charts were reviewed to determine the frequency of new mental illness diagnoses among continuing and new patients. RESULTS: The screening checklist (POPR) revealed potential mental illnesses in 148/362 outpatients, of which 98% had not been identified by the physicians who had reviewed the patients' completed POPR forms. Only five new diagnoses of mental illnesses were independently made by clinics' physicians-all in follow-up (continuing) patients. CONCLUSIONS: The physicians in the two clinics did not diagnose mental illnesses in their patients, even when written checklists of self-reports were available to them. Differences in staff attitudes may influence the data collection process, and patients' as well as physicians' responses. A weakness of this study is that the POPR might generate false positive results. Even if this were the case, the rate of previous diagnoses was still extremely low. PMID- 15825585 TI - Obsessive-compulsive disorder following brain injury: a review. AB - OBJECTIVE: This article reviews obsessive-compulsive disorder (OCD) following acquired brain injury. OCD type symptomatology without any other cognitive, behavioral, or emotional problems following a brain injury appears to be rare. However, the first onset of obsessions and compulsions following a brain injury has been described, but cognitive impairments can potentially color the presentation. METHOD: This article reviews obsessive-compulsive disorder (OCD) after brain injury, including traumatic brain injury, cerebrovascular accidents, brain tumors and brain infections. Overlapping symtomatology between OCD and brain injury is discussed. RESULTS: Findings regarding the neuro-imaging of OCD after brain injury are reviewed and integrated with neuropsychological data from studies with idiopathic OCD. CONCLUSION: There appears to be some convergance between findings from studies of the neuropsychology of idiopathic OCD and neuro imaging of OCD following brain injury. PMID- 15825586 TI - Kallmann's syndrome and schizophrenia. AB - OBJECTIVE: Kallmann's Syndrome is a heritable disorder characterized by the association of hypogonadotropic hypogonadism and anosmia or hyposmia. A common pathogenesis for KS and schizophrenia had been proposed based on shared pathologies of these two disorders, although no such clinical associations have ever been reported. METHOD: We report a 35 year old man with schizophrenia and Kallmann's Syndrome. The patient presented with signs and symptoms of hypogonadism, severe hyposmia and normal endocrine functions of the anterior pituitary. Hyposmia has been attributed to the absence of the olfactory bulbs and tracts and atrophy of the olfactory gyri, but normal olfactory mucosa. The patient presented with paranoid schizophrenia with persecutory delusions, auditory hallucinations, thought disorder, depersonalization, and gradual but marked global deterioration. RESULTS: Psychiatric evaluation revealed an entirely different psychopathological and personality profile between the patient and the six other Kallmann patients studied. Cycle sequencing analysis revealed a normal sequence of all 14 exons of the KAL gene. In conclusion, based on the presented case, Kallmann's Syndrome and schizophrenia represent a rare clinical association rather than a syndrome with a common pathogenesis, which if present should be confined to the olfactory dysfunction. PMID- 15825587 TI - Depression in children and adolescents with cystic fibrosis: case studies. AB - OBJECTIVE: Cystic fibrosis is an autosomal recessively inherited multi-system disorder of children and adults characterized by the obstruction and infection of airways presenting as a wide variety of symptoms. It has varying degrees of psychological impact on patients and their families, requiring numerous adjustments in their life styles. Newer sophisticated medical treatments of cystic fibrosis has led to an increase in life expectancy; however, the same kind of progress has not been made in the psychological interventions available to treat emotional disturbances. METHOD: We report three cases of children and adolescents with cystic fibrosis who were diagnosed with depression and treated with a combination of antidepressant medications and other psychosocial interventions. RESULTS AND CONCLUSIONS: Two of the three cases suggest that treatment with antidepressant medications has a positive influence on psychological outcomes. We further propose clinical studies to test efficacy, safety, and long-term effects of these medications in patients with cystic fibrosis. PMID- 15825588 TI - Melanocortin signalling, single nucleotide polymorphism, and eating disorders. PMID- 15825589 TI - Life and death politics. The Schiavo case is just the latest front in a much nastier war. PMID- 15825590 TI - Wrestling with the final call. When it comes to end-of-life decisions, taking an ethical path isn't always easy. PMID- 15825591 TI - Inside Terri's brain. She's probably not in pain. Still, doctors can't read her mind. PMID- 15825592 TI - When life is on the line. PMID- 15825593 TI - No, it wasn't a cynical ploy. PMID- 15825594 TI - Of birds and men. A deadly virus is brewing in Asia. Could this be the next killer pandemic? PMID- 15825595 TI - A regrettable limit on life. PMID- 15825596 TI - A time for uncertainty. PMID- 15825597 TI - Mending migraines. Can plugging a hole in the heart end headache agony? PMID- 15825598 TI - End of the affair. PMID- 15825599 TI - Society pays debts. PMID- 15825600 TI - The pressure mounts. Mental illness among grad students is on the rise. Can schools help them? PMID- 15825601 TI - A new kind of ministry. Divinity schools teach students to bring their faith to careers outside the clergy. PMID- 15825602 TI - Special report: America's best graduate schools. Schools of medicine. PMID- 15825603 TI - Not older, but faster. PMID- 15825604 TI - What do physicians want from health plans? PMID- 15825605 TI - Going on the offensive against defensive medicine. PMID- 15825606 TI - Current billing system fosters error, dishonesty. PMID- 15825607 TI - Family practice base salary continues to improve. PMID- 15825608 TI - HSAs: early returns are in. PMID- 15825609 TI - Does your managed Medicare plan have enough geriatricians? PMID- 15825610 TI - Direct-to-Consumer ads garner mixed outcomes. PMID- 15825611 TI - This country cannot continue with the system we now have. PMID- 15825612 TI - Kaiser's asthma outcomes will take your breath away. PMID- 15825613 TI - ED drugs covered, but managed care unmoved. PMID- 15825614 TI - Transparency catches eye in coalition's deal with Medco. PMID- 15825615 TI - Aptamers: slowing progression of AMD. PMID- 15825616 TI - Costly information system eventually pays off. PMID- 15825617 TI - Practice makes perfect. PMID- 15825618 TI - Harmonisation of control of use legislation for veterinary medicines. PMID- 15825619 TI - Specialist referral--the winds of change. PMID- 15825620 TI - Registration for overseas veterinarians. PMID- 15825621 TI - Enzootic pneumonia of pigs--a diagnostic dilemma. AB - In an enzootic pneumonia-free Australian pig herd, an outbreak of a severe respiratory disease in the grow-out herd was initially diagnosed as acute tracheitis and pneumonia precipitated by the dusty environment, with a superimposed mixed infection of Pasteurella multocida and Arcanobacterium pyogenes. Culture for Mycoplasma hyopneumoniae, Salmonella sp and fungi was negative. The outbreak persisted. Subsequently, gross lesions consistent with enzootic pneumonia occurred, but histological lesions were equivocal and definitive tests for M hyopneumoniae remained negative. Eighteen months after the initial outbreak, gross and histological lesions were consistent with enzootic pneumonia but serological tests were still negative. Almost 2 years later, one of four nasal swabs was positive by the polymerase chain reaction test for M hyopneumoniae, and then lung samples were sporadically positive. The pneumonic disease became endemic in the herd. Gross lesions consistent with enzootic pneumonia occurred in another herd belonging to the same company nearly 2 years after the initial outbreak. Again, results of laboratory tests were inconsistent. Despite sporadic positive polymerase chain reaction tests for M hyopneumoniae, the respiratory disease resolved within 4 months and there has been no clinical evidence of enzootic pneumonia during the subsequent 4 years. These cases raise important questions about the role of the diagnostic tests and their interpretation, and the ecology of M hyopneumoniae and its role in enzootic pneumonia. PMID- 15825622 TI - What is your diagnosis? Right forelimb lameness in an 8-year-old Rottweiler. Osteogenic sarcoma. PMID- 15825624 TI - Surgical repair of a cleft soft palate in an alpaca. AB - A 26-day-old entire male alpaca was referred for surgical treatment of a cleft soft palate. Mandibular symphysiotomy and three-layer closure of the palatal defect resulted in primary intention healing. Despite complications related to anaesthesia the alpaca made a full recovery. PMID- 15825623 TI - Fibrocartilaginous embolism in a chondrodystrophoid breed dog. AB - An 8-year-old intact male Shih Tzu dog was admitted with acute-onset tetraplegia. Magnetic resonance imaging showed an abnormality of the cervical intramedullary spinal cord. Histopathological examination of a spinal biopsy confirmed the presence of intravascular cartilaginous emboli. This is the first report of an antemortem diagnosis of fibrocartilaginous embolism in a chondrodystrophoid breed dog. PMID- 15825625 TI - Simultaneous occurrence of epitheliogenesis imperfecta with syndactyly in a calf and a lamb. PMID- 15825626 TI - The safety assessment of chemical residues in animal-derived foods. PMID- 15825627 TI - Residues of spinosad in the tissues of sheep after aerosol treatment of blowfly myiasis. AB - OBJECTIVE: To measure the residues of spinosad and chlorhexidine in the tissues of sheep after treatment of blowfly strike. PROCEDURE: Fourteen sheep with natural myiasis and 12 with artificial infestations of Lucilia cuprina larvae had the wool removed over their infestations and were treated with an aerosol wound dressing containing spinosad and chlorhexidine. Sheep were killed up to 14 days after treatment and residues of the chemicals measured in tissues. RESULTS: Chlorhexidine was not detected in any tissue. Residues of spinosad were highest in fat, lowest in muscle and intermediate in liver and kidney. The highest residue detected was 0.2 mg/kg spinosad in perirenal fat 7 days after generous treatment of a sheep with a large fly strike. Residues of spinosad in fat peaked 3 to 7 days after treatment and 1 to 3 days after treatment in liver and kidney. CONCLUSION: These studies present a realistic worst-case in struck sheep and at the highest dose studied, equivalent to 5.8 mg spinosad per kg body weight, the maximum residue detected of 0.2 mg/kg in peri-renal fat was 20% of the Australian maximum residue limit. Muscle, liver and kidney residues of spinosad were also below the Australian maximum residue limits at all times. PMID- 15825628 TI - The timing and distribution of strains around the surface of the midshaft of the third metacarpal bone during treadmill exercise in one Thoroughbred racehorse. AB - OBJECTIVE: To confirm that the midshaft dorsal cortex of the third metacarpal bone experienced higher compressive strains during fast exercise than the medial or lateral cortices, and that the strain peak occurred earlier in the hoof-down phase of the stride on the dorsal cortex than the medial or lateral cortices. DESIGN: Observations of a single horse. PROCEDURE: Strains were collected from a single, sound, 3-year-old Thoroughbred mare during treadmill exercise from rosette strain gauges implanted onto the medial, lateral and dorsal surfaces of the midshaft of the right cannon bone, simultaneously with data from a hoof switch that showed when the hoof was in the stance phase. RESULTS: Peak compressive strains on the dorsal surface of the third metacarpal bone were proportional to exercise speed and occurred at about 30% of stance. Peak compressive strains on the medial surface of the non-lead limb reached a maximum at a speed around 10 m/s and occurred at mid-stance. Peak compressive strains on the lateral surface varied in timing and size between strides at all exercise speeds, but remained less than -2000 microstrains. CONCLUSIONS: The timing of peak compressive strains on the dorsal cortex suggests a relationship to deceleration of the limb following hoof impact, so the main determinants of their size would be exercise speed and turning (as shown in previous experiments). This experiment confirms data from other laboratories that were published but not discussed, that peak compressive strains on the medial surface occur at mid stance. This suggests that they are related to the support of body weight. The strains on the lateral cortex occurred at variable times so may be associated with the maintenance of balance as well as the support of body weight. Understanding the loading of the third metacarpal bone will help to determine causes of damage to it and ways in which the bone might be conditioned to prevent such damage. PMID- 15825629 TI - Reproductive performance of early postpartum Holstein cows where oestrus was synchronised following chronic GnRH agonist treatment. AB - OBJECTIVE: The objectives of this study were: to compare the recovery of follicular development in early postpartum cows that had been treated for 7, 14 or 21 d with implants containing the GnRH agonist deslorelin; to evaluate the effectiveness of human chorionic gonadotrophin (hCG) for the induction of ovulation when a follicle was at least 10 mm in diameter following implant removal; and to compare final pregnancy rates for treated cows and untreated contemporaries. PROCEDURE: Within 3 d of calving Holstein cows were allocated to receive a single subcutaneous deslorelin implant to be left in place for either 7, 14 or 21 d, or to remain untreated as controls. Every deslorelin treated cow was monitored twice weekly for 35 d to determine the interval from implant removal to resumption of ovulation using serial transrectal ultrasonography and plasma progesterone assay. An injection of 1000 IU hCG was given to induce ovulation when a follicle of at least 10 mm diameter was first observed. Oestrous cycles of every cow were synchronised to facilitate artificial insemination (Al) at the start of the seasonally concentrated Al program and resynchronised for three rounds. Pregnancy testing was performed by ultrasonography 13 weeks after the first round of Al. RESULTS: Deslorelin implants inhibited ovulation for at least 10 d after they were removed. Ovarian follicles were smaller for the group that had implants for 21 d at the time of implant removal. Eighteen cows selected for treatment with hCG ovulated and formed multiple corpora lutea within 7 d. There was no effect of treatment duration on final pregnancy rates. After three rounds of AI the pooled final pregnancy rate for every cow that had received a deslorelin implant was similar to the rest of the herd (67% versus 63%; Deslorelin versus Herd, P > 0.1). The interval from start date of the AI program to conception was also unaffected by treatment (9.6 +/- 3.0 versus 14.8 +/- 1.7 d; Deslorelin versus Herd; P > 0.1). CONCLUSION: No significant effect was detected on the interval from implant removal to first ovulation by altering the duration of deslorelin treatment. Treatment with hCG when a follicle at least 10 mm in diameter was present induced ovulation in most cases. Although no significant improvement in fertility was found, a larger field trial using this model for induced anoestrous is necessary before any effect on fertility could confidently be stated. PMID- 15825630 TI - Peracute ammonia toxicity: a consideration in the pathogenesis of Phalaris aquatica 'Polioencephalomalacia-like sudden death' poisoning of sheep and cattle. PMID- 15825631 TI - Drug utilization: using objective data to achieve cost-efficiency. AB - By grading pharmacy benefit managers' (PBMs') performance using an objective evaluation process, employers can identify specific opportunities and establish measurable goals to define efficient drug utilization. Many employers come away from the grading process with a similar conclusion--Increased generic dispensing is a must for achieving delivery of an affordable prescription drug benefit. By focusing on the conditions that affect their population, employers are ensuring that PBMs implement practical programs that make it easy for physicians to prescribe affordable high-value drugs. PMID- 15825632 TI - Combating employee benefit cost and control issues: the case for coalition purchasing. AB - Growing in popularity, employee benefit coalitions can impact an employer's bottom line and are becoming a market factor to be dealt with in every industry and business setting. This article examines how modern-day coalitions are bringing classic aggregation theory into the 21st century and the reasons behind the growing popularity of employee benefit coalitions. It also suggests several strategic initiatives that human resource (HR) and benefit managers can take within their own organization to unlock the power of the coalition movement toward a better, more efficient buying model. PMID- 15825633 TI - Spousal surcharge programs--an overview. AB - Spousal surcharge programs help employers whose goal is to provide "above average" health benefit plans by limiting the potential "financial leakage" liabilities from covering the spouses of employees who are eligible for other health care coverage. Spousal surcharge programs are just one alternative available to help employers manage the rising cost of providing health care coverage to dependents. This article explores the prevalence, plan design, financial implications, administrative and other considerations in implementing a spousal surcharge program. PMID- 15825634 TI - Administrative Committee of the Wal-Mart Stores, Inc. Associates' Health and Welfare Plan v. Varcoi. PMID- 15825635 TI - Eat this now! How savvy marketing is contributing to the nation's obesity epidemic. PMID- 15825636 TI - No, it's not just your sweet tooth. PMID- 15825637 TI - Dances with fruit flies. PMID- 15825638 TI - Analysis of the sponge [Porifera] gene repertoire: implications for the evolution of the metazoan body plan. AB - Sponges [phylum Porifera] form the basis of the metazoan kingdom and represent the evolutionary earliest phylum still extant. Hence, as living fossils, they are the taxon closest related to the hypothetical ancestor of all Metazoa, the Urmetazoa. Until recently, it was still unclear whether sponges are provided with a defined body plan. Only after the cloning, expression and functional studies of characteristic metazoan genes, could it be demonstrated that these animals comprise the structural elements which allow the sponge cells to organize themselves according to a body plan. Adhesion molecules involved in cell-cell and cell-matrix interactions have been identified. Among the cell-cell adhesion molecules the aggregation factor (AF) is the prominent particle. It is composed of a core protein that is associated with the adhesion molecules, a 36 kDa as well as a 86 kDa polypeptide. A galectin functions as a linker of the AF to the cell-membrane-associated receptor, the aggregation receptor (AR). The most important extracellular matrix molecules are collagen- and fibronectin-like molecules. These proteins interact with the cell-membrane receptors, the integrins. In addition, a neuronal receptor has been identified, which--together with the identified neuroactive molecules--indicate the existence of a primordial neuronal network already in Porifera. The primmorph system, aggregated cells that retain the capacity to proliferate and differentiate, has been used to demonstrate that a homeobox-containing gene, Iroquois, is expressed during canal formation in primmorphs. The formation of a body plan in sponges is supported by skeletal elements, the spicules, which are composed in Demospongiae as well as in Hexactinellida of amorphous, noncrystalline silica. In Demospongiae the spicule formation is under enzymic control of silicatein. Already at least one morphogen has been identified in sponges, myotrophin, which is likely to be involved in the axis formation. Taken together, these elements support the recent conclusions that sponges are not merely nonorganized cell aggregates, but already complex animals provided with a defined body plan. PMID- 15825639 TI - Sponge-associated bacteria: general overview and special aspects of bacteria associated with Halichondria panicea. AB - Increasing evidence is accumulating that highlights the important role of bacteria in bacteria-sponge associations. It appears to be equally important to analyse the specific association of bacteria with sponges, to realise the biological function of biologically active substances produced by sponge associated bacteria, and to consider the relationship between bacteria and sponges in the search for new pharmaceutical products. In this chapter the current knowledge on bacteria-sponge associations is briefly reviewed. Results are summarised that were obtained by three major methodological approaches: (1) classical microscope observations, (2) investigations attempting to characterise sponge-associated bacteria by describing pure culture isolates, and (3) the rapidly growing evidence from genetic analyses of sponge-associated bacteria. Special emphasis is given to the evidence of possible symbiotic interactions between bacteria and sponges and to the synthesis of natural products by bacteria isolated from or associated with marine sponges. Case studies including morphological and genetic studies together with results from pure culture studies have been performed with bacteria from the sponges Rhodopaloeides odorabile, Aplysina cavernicola, and Halichondria panicea. In addition, new results on bacteria associated with Halichondria panicea are also presented. PMID- 15825640 TI - Microbial diversity of marine sponges. AB - The recent application of molecular microbial ecology tools to sponge-microbe associations has revealed a glimpse into the biodiversity of these microbial communities, that is considered just 'the tip of the iceberg'. This chapter provides an overview over these new findings with regard to identity, diversity and distribution patterns of sponge-associated microbial consortia. The sponges Aplysina aerophoba (Verongida), Rhopaloeides odorabile (Dicytoceratida) and Theonella swinhoei (Lithistida) were chosen as model systems for this review because they have been subject to both, cultivation-dependent and cultivation independent approaches. A discussion of the microbial assemblages of Halichondriapanicea is presented in the accompanying chapter by Imhoff and Stohr. Considering that a large fraction of sponge-associated microbes is not yet amenable to cultivation, an emphasis has been placed on the techniques centering around the 16S rRNA gene. A section has been included that covers the potential of sponge microbial communities for drug discovery. Finally, a 'sponge-microbe interaction model' is presented that summarizes our current understanding of the processes that might have shaped the community structure of the microbial assemblages within sponges. PMID- 15825641 TI - Full absolute stereostructures of natural products directly from crude extracts: The HPLC-MS/ MS-NMR-CD 'triad'. AB - This chapter deals with an efficient methodology available in our Center of Excellence, BIOTECmarin: the novel analytical 'triad' HPLC-MS/MS-NMR-CD. By this method, which was, in this complemented form, first introduced into phytochemical research by our group, we can not only rapidly identify known structures, but can also investigate new metabolites and establish their full absolute stereostructures online, directly from crude extracts, without the necessity of first isolating the compounds. The LC-CD option, which we have been using for the first time in natural products analysis, becomes even more valuable by the possibility of interpreting the online CD spectra by their simulation or prediction through quantum chemical calculation, thus avoiding the usual, often risky, empirical comparison with the CD spectra of (sometimes not so related) compounds of known absolute stereostructure or the application of (sometimes not really applicable) likewise empirical CD rules. The hyphenated analytical methods are additionally complemented by our synthetic expertise, again involving new concepts and strategies developed in our group. The methods and their application will first be explained and exemplified for plant-derived ('terrestrial') natural products, for which they were initially developed, and will then be applied to the online structural elucidation of novel natural products from marine organisms. PMID- 15825643 TI - Sustainable use of marine resources: cultivation of sponges. AB - Among all metazoan phyla, sponges are known to produce the largest number of bioactive compounds, some of them metabolites with human therapeutic value. Therefore, an increasing interest in basic cell biology research up to biochemical engineering can be observed aiming at the production of sponge metabolites under completely controlled conditions. One major obstacle is the limited availability of larger quantities of defined sponge material--the so called supply problem. In this chapter, different approaches used so far for producing sponge biomass by in situ aquaculture as well as some significant progress in the maintenance of sponges in aquaria are reviewed. These approaches are mainly based on old methods for producing commercial bath sponges as well as on experience in maintaining sponges in public aquaria and on the usage of artificial substrates for a natural-like colonization structure. In recent years, great efforts have been made to set up in vitro culture systems for the cultivation of sponge cells. One of the major advantages of cell cultures is the possibility to control and manipulate the cultivation conditions depending on the sponge species and the target metabolite. Up to now, monolayer cultures of dissociated sponge cells have been shown in a few cases to produce the desired product. However, to date, no continuously growing sponge cell line has been established. Organotypic culture systems, which maintain or mimic the natural tissue structure, have been developed in recent years and demonstrate a promising way towards the biotechnology of sponges. Successful attempts to produce sponge metabolites using the three-dimensional growing primmorphs are given. The use of sponge fragments, another three-dimensional approach, has reappeared and has also been successfully used as an in vitro approach as well as for the biotechnological production of boreal sponge tissue. PMID- 15825642 TI - Bioactive natural products from marine invertebrates and associated fungi. AB - Marine natural products with their unique structural features and pronounced biological activities continue to provide lead structures in the search for new drugs from nature. Invertebrates such as sponges, tunicates, mollusks and others that are either sessile or slow moving and mostly lack morphological defense structures have so far provided the largest number of marine-derived secondary constituents including some of the most interesting drug candidates. This review highlights recent research findings of our group related to natural products from marine invertebrates. Areas that are covered include ecological functions of secondary constituents from sponges against predatory fish, the search for new pharmacologically active constituents from sponges and tunicates, and sponge associated fungi as an evolving source for new bioactive natural products. PMID- 15825644 TI - Sustainable production of bioactive compounds from sponges: primmorphs as bioreactors. AB - Sponges [phylum Porifera] are a rich source for the isolation of biologically active and pharmacologically valuable compounds with a high potential to become effective drugs for therapeutic use. However, until now, only one compound has been introduced into clinics because of the limited amounts of starting material available for extraction. To overcome this serious problem in line with the rules for a sustainable use of marine resources, the following routes can be pursued; first, chemical synthesis, second, cultivation of sponges in the sea (mariculture), third, growth of sponge specimens in a bioreactor, and fourth, cultivation of sponge cells in vitro in a bioreactor. The main efforts to follow the latter strategy have been undertaken with the marine sponge Suberites domuncula. This species produces compounds that affect neuronal cells, such as quinolinic acid, a well-known neurotoxin, and phospholipids. A sponge cell culture was established after finding that single sponge cells require cell-cell contact in order to retain their telomerase activity, one prerequisite for continuous cell proliferation. The sponge cell culture system, the primmorphs, comprises proliferating cells that have the potency to differentiate. While improving the medium it was found that, besides growth factors, certain ions (e.g. silicate and iron) are essential. In the presence of silicate several genes required for the formation of the extracellular matrix are expressed (silicatein, collagen and myotrophin). Fe3+ is essential for the synthesis of the spicules, and causes an increased expression of the ferritin-, septin- and scavenger receptor genes. Furthermore, high water current is required for growth and canal formation in the primmorphs. The primmorph system has already been successfully used for the production of pharmacologically useful, bioactive compounds, such as avarol or (2'-5')oligoadenylates. Future strategies to improve the sponge cell culture are discussed; these include the elucidation of those genes which control the proliferation phase and the morphogenesis phase, two developmental phases which the cells in primmorphs undergo. In addition, immortalization of sponge cells by transfection with genomic DNA appears to be a promising way, since recent studies underscore the applicability of this technique for sponges. PMID- 15825645 TI - Approaches for a sustainable use of the bioactive potential in sponges: analysis of gene clusters, differential display of mRNA and DNA chips. AB - In recent years, analyses of the genome organization of marine sponges have begun that have led to the elucidation of selected genes and gene arrangements that exist in gene clusters (e.g. the receptor tyrosine kinase cluster and the allograft inflammatory factor cluster). Most of these studies were performed with the demosponge Suberites domuncula; but Geodia cydonium (Demospongiae), Aphrocallistes vastus (Hexactinellida) and Sycon raphanus (Calcarea) were also investigated. Both S. domuncula and G. cydonium possess a surprisingly large genome of approximately 1.7 pg DNA per haploid set. Taking the high gene density in these sponges into account and considering that predominantly single-copy DNA exists, the gene number of S. domuncula and G. cydonium was estimated to be approximately 300,000. Presumably, the large gene number in the sponge genome is due to regional gene duplication; so far evidence for a transposition in sponges has been presented. Data indicate that only 0.25 % of the total sponge genome comprises CA/TG microsatellites, and until now also no SINEs/transposable elements have been identified. Due to the rapid progress in the field of molecular biology of sponges the application of sponge genes for expression studies by DNA-array techniques (microarray) has become possible. These achievements will be further supported by the systematic analysis of the expressed genome of sponges; the results will be (partially) released (http://spongebase.uni-mainz.de/cgi-bin/blast/blastserver.cgi). In our efforts employing the results from the analysis of the genome to molecular biotechnology, we applied the technique of differential display of mRNA. One example, the effect of silicate on gene expression in S. domuncula, is outlined here. Future results will allow the identification of the genes involved in the synthesis of bioactive compounds from sponges [Porifera]. This progress will contribute considerably to a fruitful and fast development in the field of molecular marine biotechnology. PMID- 15825646 TI - Sorbicillactone A: a structurally unprecedented bioactive novel-type alkaloid from a sponge-derived fungus. AB - This chapter deals with the discovery of sorbicillactone A, as an illustrative example of the fruitful cooperation within BIOTECmarin--its isolation and chemical characterization, and its biological activities. Sorbicillactone A was isolated from a strain of Penicillium chrysogenum cultured from a sample of the Mediterranean sponge Ircinia fasciculata; it possesses a unique bicyclic lactone structure, seemingly derived from sorbicillin. Among the numerous known sorbicillin-derived structures, it is the first found to contain nitrogen and thus the first representative of a novel type of 'sorbicillin alkaloids', apparently originating from a likewise remarkable biosynthesis. Furthermore, the compound exhibits promising activities in several mammalian and viral test systems, in particular a highly selective cytostatic activity against murine leukemic lymphoblasts (L5178y) and the ability to protect human T cells against the cytopathic effects of HIV-1. These properties qualify sorbicillactone A or one of its derivatives for animal and (hopefully) also future therapeutic human trials. PMID- 15825647 TI - Controversial prompts system dumped over 'target' concerns. AB - The controversial 472 prompts proposed for the new NHS ratings system have been ditched because of fears they could turn into a set of must-do targets, the Healthcare Commission announced today. During 2005-06 the commission will concentrate on measuring performance against 24 core standards describing basic levels of performance. PMID- 15825648 TI - DoH sweetener for APMS deals. AB - Reluctant high-street pharmacy chains are to be tempted into providing primary care services through a new 'standardised' procurement programme. A central Department of Health team with a budget of 4m pounds sterling will 'pump-prime' procurement by providing advice on contract specifications and how to procure services. PMID- 15825650 TI - On extending payment by results. PMID- 15825649 TI - A children's champion with all to play for. AB - Professor Al Aynsley-Green's appointment as England's first children's commissioner takes health and social services into uncharted territory. Do events in the other countries of the UK, which already have commissioners, hold any clues on the likely impact of the appointment in England? Trevor Philpott assesses the likely direction of travel. PMID- 15825651 TI - Bursting out all over. AB - Last week the National Institute for Clinical Excellence merged with the Health Development Agency, boosting the profile of an organisation that chief executive Andrew Dillon claims 'all three parties are keen to maintain' and which 26 per cent of the population has heard of. But is the newly expanded NICE ready--and able--to deliver on its public health remit? PMID- 15825652 TI - Election campaign. Poll position. PMID- 15825653 TI - On clinical management. PMID- 15825654 TI - Infection control. A bug's life. AB - Everybody agrees that infection control is a serious problem and that something needs to be done about it, but nobody can agree exactly what. In this week's special report, we look at NHS plans to improve the worst MRSA record in Europe. Our Good Management special focuses on improved understanding of how to deal with clinical wastes. Plus John Williams comes to the defence of the private cleaning contractors blamed for the spread of MRSA. PMID- 15825655 TI - Infection control special. Good management. Expanding waste lines. AB - New waste regulations are largely incompatible with current healthcare. There are questions over how members of staff will be able to classify waste. The regulations also pose logistical problems and space constraints. PMID- 15825656 TI - Drug costs. Antibiotic pharmacist saves 10 pounds sterling per patient per day. PMID- 15825657 TI - Training. Stopping the use of dangerous disinfectants. PMID- 15825658 TI - On why politics and MRSA don't mix. PMID- 15825659 TI - Campaigns. How to create a 'clean culture'. PMID- 15825660 TI - Patient involvement. Harness the public to improve cleanliness. PMID- 15825661 TI - Mental health services. Must try harder. AB - Professor Louis Appleby's review sets out progress on mental healthcare reform, and future priorities. Spending is higher than ever, but there is still a lack of transparency around funding. More must be done to meet the needs of black and minority ethnic communities. PMID- 15825662 TI - Building. Design for lives. PMID- 15825663 TI - Finance. Money changes everything. AB - The DoH wants to introduce payment by results to mental healthcare by 2008-09. The NHS Information Authority case-mix service is developing healthcare resource groups for inpatients and community care. One way to introduce payment by results to the sector would be to pay per item of service, such as per inpatient day. PMID- 15825664 TI - Good management. The right side of bed. AB - A mental health trust is tackling out-of-area care by reducing available beds. Its recovery model of care reduces the reliance on hospital care. Out-of-area treatment has dropped to zero and bed usage is at 84 percent. PMID- 15825665 TI - Benefits. Help inpatients claiming benefits. PMID- 15825666 TI - Prescribing. Switching schizophrenia treatments. PMID- 15825667 TI - Recruitment. Lessons on setting up primary care mental health teams. PMID- 15825668 TI - Care management. Treating personality disorder. PMID- 15825669 TI - Dealing with staff concerns. 'Print this if you dare'. PMID- 15825671 TI - Staff incentives. How to say thank you. PMID- 15825670 TI - Recruitment barometer. Primed for change. PMID- 15825672 TI - [Antidepressants and suicide]. PMID- 15825673 TI - [A new atypical antipsychotic with partial dopamine agonist effect (aripiprazole)]. AB - Aripiprazole act as a partial agonist on the D2 receptors in contrast to the other antipsychotic agents which exert a pure antagonist action. In the autoreceptors with large receptor reserve aripiprazole shows primarily agonistic action, while postsynaptically its antagonistic action becomes predominant. In addition aripiprazole is an antagonist at the 5-HT2A, receptor and a partial agonist on the 5-HT1 receptor. On the basis of these receptor actions it is suggested that aripiprazole works as a stabilizer of both the dopaminergic and serotoninergic systems. Probably due to this dual stabilizing effect aripiprazole ameliorates positive and negative as well as anxiety and cognitive symptoms. Especially the very rare extrapyramidal symptoms, prolactine level elevation, QT interval lengthening and absence of severe body weight increase and metabolic disturbances are noteworthy, which all seriously impair the health condition, the quality of life and the therapy adherence of the patients. Due to the very low affinity of aripiprazole to the H1 and muscarinic receptors aripiprazole practically does not lead to increase of body weight. The significant clinical efficacy coupled with good tolerability assures high level clinical effectiveness for aripiprazole in the broad clinical practice. PMID- 15825674 TI - [Review of antidepressants from the TCAs to the third generation drugs]. AB - Standpoints regarding the pathomechanism of unipolar major depressive disorder (MDD) and the relevant pharmaco-therapeutical possibilities are discussed in the paper. Generations of the antidepressants from the tri- and teracyclic drugs (TCAs) to the modern, dual-acting antidepressants, including the still popular selective serotonin reuptake inhibitors (SSRIs) are surveyed in more detailed. Pharmacodynamic properties of each antidepressant subgroup with the therapeutical benefits, the occasional side-effects and with some characteristic features of the detailed ingredients in certain cases are emphasized. Some benefits and possible disadvantages regarding the combinations are also discussed briefly. The main aim of this review is to give an objective approach to the psychiatrists in order to choose the best and optimal pharmaco-therapeutical method in their daily practice. PMID- 15825675 TI - Suicide prevention strategies--a brief review. AB - Since suicide is a very complex, multicausal human behaviour, its prevention should also be complex. The prediction of suicide is very difficult at the level of the general population, but it is much easier among patients with certain mental disorders, because most persons who kill themselves have diagnosable and treatable psychiatric disorders. This article reviews the most important biological and non-biological suicide prevention strategies. PMID- 15825676 TI - Treatment possibilities of Alzheimer's disease. AB - A brief summary of the current treatment of Alzheimer disease (AD) (cholinergic replacement therapy, influence of glutamatergic neurotransmission, treatment based on the beta-amyloid cascade theory, antioxidants, anti-inflammatory drugs) clearly proves that the applied strategies are practically inefficient. We describe therefore the rationale and design of a reasonable clinical trial to test the validity of Knoll's concept that the administration of a synthetic mesencephalic enhancer substance prior to the precipitation of the symptoms is our only chance to significantly reduce the prevalence of the two main neurodegenerative disorders AD and Parkinson's disease (PD). Considering that in the population over 65 there are substantial sex (68% female, 32% male) and geographical (highest rate: 10% in USA) differences in the incidence of AD, we propose to perform the clinical trial in 75-85 year old females in the USA. Individuals without (Group 1) and with (Group 2) predisposition to AD should be selected. One third in each group should be treated daily with placebo, (-) deprenyl (1 mg) and (-)-BPAP (1 mg), respectively. Series of studies proved already the protective effect of the synthetic mesencephalic enhancer substances against age-related neurodegenerative changes in the brain. We may therefore expect a significant difference in the placebo versus drug treated groups in the number of individuals who will precipitate with the passing of time the symptoms of AD or PD. The introduction of a safe and efficient prophylactic therapy that significantly decreases the prevalence of AD is a necessity which cannot be further postponed. PMID- 15825677 TI - The use of the synthetic enhancer substances (-)-deprenyl and (-)-BPAP in major depression. AB - There is still a great need for the development of antidepressants with a new pharmacological spectrum. The finding that phenylethylamine and tryptamine are endogenous enhancers of the impulse propagation mediated release of catecholamines and serotonin in the brain, and the development of synthetic mesencephalic enhancer substances opened the possibility to stimulate catecholaminergic and serotonergic neurons in the mesencephalon via a previously unknown mechanism. (-)-Deprenyl, a prototype of the phenylethylamine-derived synthetic enhancer substances, stimulates the catecholaminergic neurons in the brain but is almost ineffective on the serotonergic neurons. R-(-)-1-(benzofuran 2-yl)-2-propylaminopentane, (-)-BPAP, the recently developed tryptamine-derived selective synthetic mesencephalic enhancer substance, a hundred times more potent compound than (-)-deprenyl, acts also on the serotonergic neurons. The evaluation of the special pharmacological profile of the synthetic mesencephalic enhancer substance, especially the high potency and the unusual safety and tolerability of (-)-BPAP provide hope that this compound may in the future significantly improve the effectiveness of drug therapy in major depression and its combination with uptake inhibitors may substantially diminish the number of therapy resistant cases. PMID- 15825678 TI - [Misdiagnosed PTSD and zeldox pharmacotherapy in case of a political prisoner]. AB - Treatment of survivors of political terror is an emerging and difficult field. Reports on posttraumatic stress disorder (PTSD) in political prisoners within the former Eastern Block countries is low and mostly restricted to German sources. During the totalitarian period administrative and clinical decisions often had to take into account political realities not found in other treatment environments. That practice might have lead to biased professional training, lack of experience extending into the post-communist era and leading to current underpresentation of PTSD. The authors present a case report of a Hungarian political prisoner with long history of PTSD who had a "carry over" diagnosis of paranoid schizophrenia even 15 years after the collapse of the communist regime. After decades of continuous administration of antipsychotic and antidepressive medications, either alone or in combination, Zeldox monotherapy has proven to be the most effective treatment for this patient. PMID- 15825679 TI - The relationship between surface adsorption and the hydrolysis of amitraz in anionic surfactant solutions. AB - The adsorption of amitraz to various adsorbents was studied in terms of the amount and rate of adsorption and the effect that adsorption had on the stability of amitraz in an aqueous environment. Adsorption results showed that in terms of their ability to adsorb amitraz from solution the adsorbents tested in this study can be ordered as follows: coarse carbon > cation exchange resin > or = anion exchange resin > fine carbon. Amitraz was not adsorbed on sand and potassium oxihumate. Adding sodium lauryl sulfate and potassium oxihumate to aqueous suspensions of suspended adsorbents containing adsorbed amitraz showed that both these anionic surfactants significantly increased the hydrolysis rate because the half-lives for amitraz was reduced from 27 days for a suspension to only 8 hours for amitraz adsorbed to a cation exchange resin and suspended in an aqueous buffer pH 5.8 containing 0.5% of the anionic surfactant sodium lauryl sulfate and 12 hours when 1% potassium oxihumate was added. PMID- 15825680 TI - Photo-fenton assisted reaction of dimethoate in aqueous solutions. AB - The photo-Fenton reaction of an organophosphorus insecticide, dimethoate (O,O dimethyl methylcarbamoylmethyl phosphorodithioate), was studied by following the identification and determination of the decomposition products and the total carbon removal rate. The reactions were performed in a batch recycle reactor, at room temperature, using UV radiation, H2O2 as oxidant, and FeCl3 x 6H2O as catalyst. The oxidation results were determined with a total organic carbon (TOC) analyzer and ion chromatography. The presence of reaction products was identified by gas chromatography-mass spectrometry (GC-MS). Apart from the sulfate, phosphate, and ammonium ions, the presence of dimethyl phosphite, N-methyl acetamide, and formic acid was also detected. Excess of H2O2 concentration did not influence the reaction rate. The expression for the total carbon removal was assessed and the TOC removal rate constants were calculated. PMID- 15825681 TI - Heterogeneous photocatalytic degradation of picloram, dicamba, and floumeturon in aqueous suspensions of titanium dioxide. AB - Heterogeneous photocatalytic degradation of three-selected herbicide derivatives: (1) picloram (4-Amino-3,5,6-trichloropyridine-2-carboxylic acid, (2) dicamba (2 Methoxy-3,6-dichlorobenzoic acid, and (3) floumeturon (N,N-Dimethyl-N-[3 (trifluoromethyl)phenyl]-urea) 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 herbicide was studied under different conditions such as pH, catalyst concentration, substrate concentration, different types of TiO2, and in the presence of electron acceptors 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 photocatalysts in the case of dicamba (2) and floumeturon (3), whereas Hombikat UV100 was found to be better for the degradation of picloram (1). The herbicide picloram (1) was found to degrade faster as compared to dicamba (2) and floumeturon (3). The degradation products were analyzed by gas chromatography mass spectrometry (GC/MS) technique, and plausible mechanisms for the formation of products have been proposed. PMID- 15825682 TI - Interaction effects of insecticides on microbial populations and dehydrogenase activity in a black clay soil. AB - Three insecticides, monocrotophos, quinalphos, and cypermethrin, were applied at 0, 5, 10, and 25 microg g(-1) either singly or in combination to a black clay soil to investigate their effects on the soil microflora and dehydrogenase activity. All three insecticides significantly enhanced the proliferation of bacteria and fungi and the soil dehydrogenase activity even at the highest level of 25 microg g(-1). Monocrotophos or quinalphos in combination with cypermethrin at tested levels interacted significantly to yield additive, synergistic, and antagonistic responses toward bacteria and fungi and dehydrogenase activity in soil. Antagonistic interactions were more pronounced toward soil microflora and dehydrogenase activity when the two (monocrotophos or quinalphos + cypermethrin) insecticides were present together in the soil at highest level (25 + 25 microg g(-1)), whereas synergistic or additive responses occurred at lower level with the same combination of insecticides in soil. PMID- 15825683 TI - Effects of azadirachtin on Beet soilborne pomovirus and soil biological properties on sugar beet. AB - Beet soilborne pomovirus (BSBV) is an important soilborne virus disease in growing areas of sugar beet. BSBV was transmitted by Polymyxa betae Keskin. Effects of azadirachtin on BSBV and soil biological properties were studied under a greenhouse. The presence of BSBV was tested in soil samples using bait plant test and triple antibody sandwich-enzyme-linked immunosorbent assay (TAS-ELISA). The concentration of BSBV in sugar beet roots was significantly reduced by the application of azadirachtin to the soils. TAS-ELISA absorption values in 1.52, 3.04, and 7.60 ppm were lower than that 0.38 and 0.76 ppm of azadirachtin. Furthermore, in this study, it was determined the 0.38, 0.76, and 1.52 ppm doses of azadirachtin in soils had high amounts of soil biological properties (Cmic, BSR, DHA), while the 3.04 and 7.60 ppm doses had no statistical significance compared to the control treatments. PMID- 15825684 TI - A cost-effective screening method for pesticide residue analysis in fruits, vegetables, and cereal grains. AB - This paper reports the results of studies performed to investigate the potential of applying thin layer chromatography (TLC) detection in combination with selected extraction and cleanup methods, for providing an alternative cost effective analytical procedure for screening and confirmation of pesticide residues in plant commodities. The extraction was carried out with ethyl acetate and an on-line extraction method applying an acetone-dichloromethane mixture. The extracts were cleaned up with SX-3 gel, an adsorbent mixture of active carbon, magnesia, and diatomaceous earth, and on silica micro cartridges. The Rf values of 118 pesticides were tested in eleven elution systems with UV, and eight biotest methods and chemical detection reagents. Cabbage, green peas, orange, and tomatoes were selected as representative sample matrices for fruits and vegetables, while maize, rice, and wheat represented cereal grains. As an internal quality control measure, marker compounds were applied on each plate to verify the proper elution and detection conditions. The Rf values varied in the different elution systems. The best separation (widest Rf range) was achieved with silica gel (SG)--ethyl acetate (0.05-0.7), SG--benzene, (0.02-0.7) and reverse phase RP-18 F-254S layer with acetone: methanol: water/30:30:30 (v/v) (0.1-0.8). The relative standard deviation of Rf values (CV(Rf)) within laboratory reproducibility was generally less than 20%, except below 0.2 Rf, where the CVRf rapidly increased with decreasing Rf values. The fungi spore inhibition, chloroplast inhibition, and enzyme inhibition were found most suitable for detection of pesticides primarily for confirming their identity or screening for known substances. Their use for determination of pesticide residues in samples of unknown origin is not recommended. PMID- 15825685 TI - Dissipation of chlorothalonil, chlorpyrifos, and profenofos in a Malaysian agricultural soil: a comparison between the field experiment and simulation by the PERSIST model. AB - A comparison of dissipation of chlorothalonil, chlorpyrifos, and profenofos in a Malaysian agricultural soil between the field experiment and simulation by the PERSIST model was studied. A plot of sweet pea (Pisum sativum) from a farm in the Cameron Highlands was selected for the field experiment. The plot was treated with chlorothalonil, chlorpyrifos, and profenofos. Core soil collection was conducted according to the sampling schedule. Residues of the three pesticides were analyzed in the laboratory. Simulations of the three pesticides' persistency were also conducted using a computer-run software PERSIST. Generally, predicted data obtained using PERSIST were found to be high for the three pesticides except for one field measurement of chlorpyrifos. The predicted data for profenofos, which is the most mobile of the three pesticides tested, was not well matched with the observed data compared to chlorothalonil and chlorpyrifos. PMID- 15825686 TI - Use of composted animal manures to reduce 1,3-dichloropropene emissions. AB - Atmospheric emission of the soil fumigant 1,3-dichloropropene (1,3-D) has been associated with the deterioration of air quality in certain fumigation areas. To minimize the environmental impacts of 1,3-D, feasible and cost-effective control strategies are in need of investigation. One approach to reduce emissions is to enhance the surface layer of a soil to degrade 1,3-D. A field study was conducted to determine the effectiveness of composted steer manure (SM) and composted chicken manure (CKM) to reduce 1,3-D emissions. SM or CKM were applied to the top 5-cm soil layer at a rate of 3.3 or 6.5 kg m(-2). An emulsified formulation of 1,3-D was applied through drip tape at 130.6 kg ha(-1) into raised beds. The drip tape was placed in the center of each bed (102 cm wide) and 15 cm below the surface. Passive flux chambers were used to measure the loss of 1,3-D for 170 h after fumigant application. Results indicated that the cumulative loss of 1,3-D was about 48% and 28% lower in SM- and CKM-amended beds, respectively, than in the unamended beds. Overall, both isomers of 1,3-D behaved similarly in all treatments. The cumulative loss of 1,3-D, however, was not significantly different between the two manure application rates for either SM or CKM. The results of this study demonstrate the feasibility of using composted animal manures to control 1,3-D emissions. PMID- 15825687 TI - The effects of magnesium and ammonium additions on phosphate recovery from greenhouse wastewater. AB - Phosphorus recovery from greenhouse wastewater, using precipitation crystallization, was conducted under three levels of calcium concentration, 304 mg/L (7.6 mmol/L), 384 mg/L (9.6 mmol/L), and 480 mg/L (12 mmol/L), and also with additions of ammonium and magnesium into the wastewater. Jar test results confirmed high phosphate removal, with more than 90% of the removal achieved with a pH as low as 7.7. Under the low calcium concentration, ammonium addition affected the chemical reactions at pH lower than 8.0, where struvite was produced; when the pH was raised to 8.8, other calcium compounds dominated the precipitation. Under the medium calcium concentration, ammonium and magnesium addition helped struvite precipitation in the low pH range. Hydroxyapatite (HAP) was the main product. Under the high calcium concentration, ammonium addition showed no effects on the precipitation. PMID- 15825688 TI - Heavy metal uptake by two edible Amaranthus herbs grown on soils contaminated with lead, mercury, cadmium, and nickel. AB - The uptake of an element by a plant is primarily dependent on the plant species, its inherent controls, and the soil quality. Amaranthus hybridus (green herbs) and Amaranthus dubius (red herbs) were chosen to investigate their response and ability to accumulate and tolerate varying levels of elements in their roots and shoots. Red herbs and green herbs were grown in soil pots contaminated with three mixtures of Cd(II), Ni(II), Pb(II), and Hg(II). Plants in the control treatment were grown in the absence of the heavy metals mixture. The distribution of Cd, Ni, Pb, and Hg in the plants (in roots, stems, and leaves) was determined in two stages. Stage 1, after 5 weeks of plant growth and stage 2, full grown after 10 weeks of growth. In the red herbs the Cd concentration in the leaves at stage 2 was 150 ppm and was present in higher concentrations than Ni, Hg, and Pb. At the highest contamination level, in the green herbs plant, Hg was present in the highest concentration in the root, i.e., 336 ppm at stage 1, while the level in the leaves was 7.12 ppm. Both the green and red herbs species showed an affinity for Ni and Cd with moderate to high levels detected in the leaves, respectively. PMID- 15825689 TI - Mosquito surveys carried out on Green Island, Orchid Island, and Penghu Island, Taiwan, in 2003. AB - Field surveys of mosquitoes were carried out on Green, Orchid, and Penghu Islands in 2003 to ascertain the status of mosquito vectors. Eighteen species of mosquitoes were collected, including three species of Anopheles, four species of Aedes, eight species of Culex, two species of Armigeres, and one species of Malaya. Seventeen previously recorded species were not collected in this study but 11 species collected had not previously been recorded. Ten newly recorded species, An. maculatus, An. takasagoensis, Ae. alcasidi, Ae. lineatopennis, Ae. vexans vexans, Ar. omissus, Cx. vishnui, Cx. halifaxii, Cx. hayashii, and Cx. neomimulus, were collected on Green Island and one previously unrecorded species, Ar. subalbatus, was collected on Orchid Island. Potential vectors An. maculatus and An. sinensis, malaria vectors in Korea and Mainland China, Ae. albopictus, a vector of dengue in Taiwan and West Nile virus in the USA, Cx. vishnui and Cx. tritaeniorhynchus, Japanese encephalitis vectors in Taiwan, Ae. vexans vexans, an eastern equine encephalitis vector in the USA, and Cx. quinquefasciatus, a vector of filariasis in Taiwan and West Nile virus in the USA, were among the mosquito species collected. PMID- 15825690 TI - Mutation of the p53 tumor suppressor gene in transitional cell carcinoma of the urinary tract in Taiwan. AB - Early detection of a mutated p53 gene is thought to provide useful information in a wide range of human tumors. The aim of this study was to identify the role of the p53 gene in transitional cell carcinoma of the urinary tract. From March 1992 to July 2003, 75 patients (54 men and 21 women) with a mean age of 66.85 years and pathologically diagnosed transitional cell carcinoma were enrolled in this study. Fifty-eight patients had bladder cancer, eight had ureteral cancer, and nine had renal-pelvic cancer. Rapid screening for mutation of the p53 gene was performed using polymerase chain reaction (PCR), single-strand conformation polymorphism (SSCP), and sequencing analysis. Primer sets were designed to amplify fragments within exons 4, 5, 6, 7, and 8 of the p53 gene. Pathology classified 37 tumors as low grade and 38 as high grade. Tumor stage was pT1 or less in 29 patients and at least pT2 in 46 patients. Of the 75 patients in this study, 47 (62.7%) had a p53 mutation. Of the patients with a p53 mutation, 33 (70.2%) had invasive tumors. Invasive tumors were associated with p53 mutation (p < 0.05). Noted in 20 patients (26.7%), exon 4 was the most common site of the mutation. Of the patients with exon 4 mutations, 15 (75%) had invasive tumors and nine (45%) had high-grade tumors. Additionally, among the 20 patients with a common polymorphism at codon 72, 16 (80%) had invasive tumors and 14 (70%) had high-grade tumors. In this study, 62.7% of patients with transitional cell carcinoma had a p53 mutation, suggesting that the p53 gene mutation may be used as a marker of transitional cell carcinoma. Invasive tumors are more likely to have a p53 gene mutation. A simple analysis of the p53 gene using PCR/SSCP is suitable for screening for p53 abnormalities in transitional cell carcinoma. The relationship between cancer risk and the codon 72 polymorphism of exon 4 needs further investigation. PMID- 15825691 TI - Compurerized morphometric study of thyroid follicular carcinoma in correlation with known prognostic factors. AB - This study investigates the correlation between computer-assisted nuclear morphometry and known prognostic factors in thyroid follicular carcinoma. Thirty six patients with thyroid follicular carcinoma who underwent surgery between 1991 and 2001 were grouped according to sex, age, size of the primary lesion, the presence of vascular invasion, and metastases. Four nuclear parameters were measured and analyzed: mean nuclear area, mean nuclear perimeter, largest to smallest diameter ratio of the nuclei, and coefficient of variation of the nuclear area. Our results indicated that none of the chosen nuclear variables were significantly correlated with the prognostic factors studied. In conclusion, nuclear morphometry does not seem to correlate with known prognostic factors and cannot serve as an additional predicting factor for biologic behavior. PMID- 15825692 TI - Ultrasonographic findings in hemiplegic knees of stroke patients. AB - Clinical and radiologic asymmetric arthritic differences between paralyzed and nonparalyzed limbs of stroke patients have been reported. Arthritic pathology aggravates motor dysfunction and compromises rehabilitation. Musculoskeletal ultrasonography plays an important role in showing soft tissue and the articular cartilage of the knee. Fifty-nine patients with either ischemic or hemorrhagic stroke-induced right or left hemiplegia were recruited to evaluate soft-tissue and intra-articular cartilage changes in hemiplegic knees of stroke patients using ultrasonography. An additional 15 subjects (30 knees) without knee disease or a history of knee trauma or surgery were used as controls. There were significant differences in suprapatellar effusion and patellar tendinitis between hemiplegic and nonhemiplegic knees. Suprapatellar effusion and pes anserinus tendinitis were correlated with Brunnstrom stage. The length of time since stroke onset was not significantly correlated with positive ultrasonographic findings in hemiplegic knees. In conclusion, ultrasonography is useful for detecting periarticular soft-tissue changes and intra-articular lesions in hemiplegic knees of stroke patients. PMID- 15825693 TI - Acute respiratory distress syndrome after early successful primary percutaneous coronary intervention therapy in acute myocardial infarction: a case report. AB - Acute respiratory distress syndrome (ARDS) is characterized by acute-onset dyspnea, diffuse bilateral pulmonary infiltration, low pulmonary capillary wedge pressure (PCWP), and an arterial oxygen tension/inspired oxygen fraction (PaO2/FiO2) ratio of less than 200 mmHg. Acute myocardial infarction (AMI), whether complicated by circulatory arrest, cardiogenic shock, and hypotension or not, was reported as an etiologic factor in the development of ARDS in the prethrombolytic era. In the thrombolytic era, two cases of AMI complicated with ARDS have been reported. ARDS in these two patients resulted from anaphylactic reaction to the thrombolytic agent and not from the hemodynamic consequences of AMI. Development of ARDS during the AMI period has not been reported after early successful primary percutaneous coronary intervention (PCI). Herein, we report a 61-year-old male patient with persistent chest pain who was diagnosed with Killip II anterior ST-segment elevation AMI. He was treated successfully with primary PCI 2.5 hours after the onset of chest pain. Unfortunately, on the third hospital day, acute-onset dyspnea (respiratory rate, 33 beats/min), fever (38.5 degrees C), leukocytosis (white blood cell count, 18,360/microL), and diffuse bilateral pulmonary infiltration were noted. ARDS was diagnosed from the low PCWP (8 mmHg) and a PaO2/FiO2 of less than 200 mmHg (160 mmHg). No usual causes of ARDS such as infection, aspiration, trauma, shock, or drug reactions were noted. We assumed that, in this particular patient, the systemic inflammatory response syndrome frequently induced by AMI might have caused this episode of ARDS. This may imply that AMI itself is a possible etiology of ARDS. PMID- 15825694 TI - Ectopic pancreatic tissue presenting as an umbilcal mass in a newborn: a case report. AB - The case of an ectopic pancreatic mass at the umbilicus in an 8-day-old male neonate is reported, the youngest patient with this condition ever reported in an English-language journal. The patient was healthy except for a protruding mass with intermittent mucous discharge at the base of the umbilical stump. Surgical intervention was performed under the impression of the umbilical mass. Pathology diagnosed an ectopic pancreas with acute hemorrhage. To the best of our knowledge, only one case of ectopic pancreas presenting as an umbilical mass with intermittent mucous discharge has previously been reported. PMID- 15825695 TI - Treatment of laryngeal radionecrosis with hyperbaric oxygen therapy: a case report. AB - An 81-year-old male with early-stage laryngeal carcinoma had been treated with 60 Gy curative radiotherapy. He complained of a sore throat, foul odor in the mouth, progressive dyspnea, and fever 2 months after the completion of radiotherapy. Direct laryngoscopy revealed narrowing of the glottis with diffuse ulcerative necrotic tissue. Biopsies at multiple sites and pathology revealed intense coagulation necrosis with complete denudation of covering epithelium without any malignancy. Since laryngeal radionecrosis was suspected, the patient received hyperbaric oxygen (HBO) therapy 40 times for 1 hour of 100% O2 at 2 atm absolute pressure. His clinical symptoms gradually improved and repeated endolaryngeal biopsies were undertaken near the end of HBO therapy and again 6 months later. The patient's larynx healed completely with diffuse fibrosis and no malignant cells were found on pathology. Radionecrosis must be differentiated from cancer recurrence following curative radiotherapy for early laryngeal cancer. HBO therapy could be a useful treatment adjunct for laryngeal radionecrosis. PMID- 15825696 TI - Intravascular lymphomatosis mimicking acute disseminated encephalomyelitis: a case report. AB - We describe the clinical, radiologic, and postmortem findings in a 49-year-old woman with intravascular lymphomatosis. The patient presented with progressive limb weakness followed by progressive disseminated cerebral neurologic symptoms. Disseminated encephalomyelitis was suspected due to the clinical and radiologic findings. Steroid pulse therapy and intravenous immunoglobulin were given but did not help. The patient died of multiple organ failure 3 months after the onset of symptoms. A diagnosis of disseminated intravascular large B-cell lymphomatosis was established based on findings from histopathology and immunohistochemistry studies on autopsy specimens of the brain and other visceral organs collected postmortem. PMID- 15825697 TI - Lower urinary tract symptoms in Thai women attending the menopause clinic: prevalence and associated factors. AB - OBJECTIVE: To determine the prevalence of lower urinary tract symptoms and associated factors in women attending the menopause clinic. METHOD: Nine hundred and fifty-six women attending the menopause clinic, Ramathibodi Hospital were interviewed regarding their general health issues and lower urinary tract symptoms by means of an anonymous questionnaire. Demographic data, obstetric history, and underlying diseases were analysed by using Student t-test, Chi square and Fisher exact test. P < 0.05 was considered as a level of significance. RESULTS: A total of 956 women, mean age 52.89 +/- 5.80 years, completed the questionnaire. The prevalence of stress incontinence, nocturia, urgency, frequency, and urge incontinence were 58.3%, 40.3%, 33.9%, 22.7%, and 6.6%, respectively. Lower urinary tract symptoms was found to be associated with marital status, coexisting medical diseases, menopausal status, previous term delivery, and vaginal delivery (P < 0.05). CONCLUSIONS: Lower urinary tract symptoms was a common problem among women attending the menopause clinic. Marital status, coexisting medical diseases, menopausal status, parity, and mode of delivery were associated with this problem. PMID- 15825698 TI - Accuracy of Nugent's score and each Amsel's criteria in the diagnosis of bacterial vaginosis. AB - OBJECTIVE: To determine the diagnostic accuracy of Nugent's score and each Amsel's criterion in the diagnosis of bacteria vaginosis (BV), considering Amsel's criteria as the gold standard. DESIGN: Cross-sectional, descriptive study (diagnostic test) Setting: Family planning clinic, Siriraj Hospital, Mahidol University. SUBJECTS: A total of 217 women who attended the Family Planning Clinic at Siriraj Hospital between August and December 2003. METHOD: Pelvic examination was performed on each participant. Samples of vaginal discharge was tested for BV infection using both Amsel's criteria and Nugent's score. Interpretation was made blinded without knowledge of each test result. Using Amsel's criteria as a gold standard, sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of Nugent's score and each of Amsel's criteria were estimated. RESULTS: Considering Amsel's criteria as the gold standard, Nugent's score showed a sensitivity of 65.6% (95%CI 46.8%, 80.8%), specificity of 97.3% (95%CI 93.5%, 99.0%), positive predictive value (PPV) of 80.8% (95%CI 60.0%, 92.7%), negative predictive value (NPV) of 94.2% (95%CI 89.7%, 96.9%) and accuracy of 92.6% (95%CI 88.1%, 95.6%). Both vaginal pH and whiff test demonstrated 100% sensitivity. However, vaginal pH showed lower specificity than the whiff test (58.9% and 97.3% respectively). CONCLUSION: Nugent's score might not be suitable to use as a screening test for diagnosis of BV due to its low sensitivity. The whiff test is the best clinical criteria of Amsel's criterion in the diagnosis of BV due to its high sensitivity and specitivity. PMID- 15825699 TI - Maternal grief after abortion and related factors. AB - OBJECTIVE: To investigate maternal grief after abortion and the factors that might relate to the intensity of maternal grief. DESIGN: Cross-sectional, descriptive study. SUBJECT: 132 women who attended the abortion clinic, Department of Obstetrics and Gynecology, Faculty of Medicine, Siriraj Hospital, Thailand. METHOD: The subjects were asked to complete the questionnaires including demographic characteristics, history of previous pregnancy, and Perinatal Grief scales, two weeks after abortion. The data was then analyzed to determine maternal grief and related factors of grief intensity. RESULTS: There were 7 women with severe grief intensity (5.3%), 50 with moderately grief intensity (37.9%) and 75 with mild grief intensity (56.8%). The factors associated with PGS scores were low income, had had ultrasonography, gestational age of > 16 weeks and methods of treatment. CONCLUSION: Grief is worldwide among women who have recently aborted. The related factors with grief intensity can be used for screening psychological problems of the women who experience abortion. If found, the physicians can closely observe and help them to work through their coping mechanism and prepare them to get another successful pregnancy in the future. PMID- 15825700 TI - Efficacy and safety of 12-week treatment with fenofibrate 300 mg in Thai dyslipidemic patients. AB - BACKGROUND: High levels of low density lipoprotein (LDL) cholesterol is a known major factor in atherosclerosis. In addition to LDL-cholesterol, an increase in the triglycerides-rich lipoprotein and a decrease in HDL-cholesterol increase the risk of coronary artery disease. Fenofibrate, a fibric acid derivative, is highly effective in reducing serum triglycerides and LDL-cholesterol and produces a modest increase in HDL-cholesterol. The present study was done to evaluate the efficacy of fenofibrate at 300 mg daily on serum lipid profiles and to study the drug safety and tolerability of fenofibrate in Thai patients. MATERIAL AND METHOD: Forty patients with elevated serum total cholesterol, LDL cholesterol were recruited for 12 weeks of 300 mg per day of fenofibrate therapy. Blood analysis for lipid profiles, liver function test, creatinine and muscle enzyme were done at the begining and end of the study. RESULTS: The mean baseline total cholesterol, LDL-cholesterol, triglycerides and HDL-cholesterol were 249 mg/dl, 160 mg/dl, 325 mg/dl and 43 mg/dl respectively. Significant changes of all lipid parameters from baseline were observed after 12 weeks of treatment. Reduction of serum total cholesterol, LDL-cholesterol and triglycerides were 16, 23, and 41 percent respectively. Increased serum HDL-cholesterol of 14 percent was also observed. One patient withdrew from the trial due to chest pain. Two asymptomatic elevated transaminase were detected during the study. CONCLUSION: Fenofibrate at 300 mg per day is effective and safe in treating Thai patients with dyslipidemia. PMID- 15825701 TI - Validation of the OSTA index for discriminating between high and low probability of femoral neck and lumbar spine osteoporosis among Thai postmenopausal women. AB - RATIONALE: Bone mineral density (BMD) measurement of postmenopausal women is needed to diagnose osteoporosis. This is an expensive procedure and available in only a few hospitals in Thailand. The Osteoporosis Self-Assessment Tool for Asians (OSTA) index, based on age and weight, has been developed for screening of postmenopausal Asian women to identify women for whom there is little advantage in undergoing bone densitometry. This index was developed for prediction of neck of the femur osteoporosis and did not include spinal osteoporosis, which is also common. Furthermore, the index was based on data from a sample of mainly ethnic Chinese. There is evidence that the BMD of Thai women is significantly higher for the same age and weight than that of women of several other Asian ethnics, thus there is a need to validate the appropriateness of the OSTA index for both femur and spinal osteoporosis among the Thai population. OBJECTIVE: To determine the properties of the OSTA index as a screening tool among postmenopausal women in southern Thailand. PATIENTS: Three-hundred-and-eighty-eight postmenopausal women, with no history of disease or use of drugs associated with secondary osteoporosis and no history of treatment for osteoporosis, attending the postmenopausal clinic or internal medicine out-patient department of Songklanagarind Hospital, a teaching hospital in southern Thailand, between November 2000 and April 2002. METHOD: BMD t-scores of the neck of the femur and lumbar spine were collected retrospectively and a diagnosis of osteoporosis made according to WHO criteria. Sensitivity and specificity and their 95% confidence limits were calculatedfor the dichotomized OSTA index. RESULTS: Thirty-one percent of the women were detected as having osteoporosis, comprising neck of the femur (12 percent) and lumbar spine (31 percent). Twenty nine women (7.5 percent) had evidence of spinal fracture. The OSTA index at the standard cut-point of < or = -1 had a sensitivity and specificity of 0.93 (95% CI: 0.82 - 0.99) and 0.61 (95% CI: 0.56-0.66) respectively for neck of the femur but only 0.80 (95% CI: 0.72-0.87) and 0.70 (95% CI: 0.64-0.75) respectively for lumbar spine. Raising the cut-point to < or = 0 reduced the high false negative rate (0.20) in prediction of lumbar spine osteoporosis to 0.07 (95% CI: 0.03-0.13) and identified 27 percent (95% CI: 23-32 percent) of all women at low risk of osteoporosis at either site. These women may not need to undergo BMD measurement. CONCLUSION: The standard cut-point of the OSTA index could identify most Thai postmenopausal women with osteoporosis of the neck of the femur. However, to improve the sensitivity of detection of osteoporosis of the lumbar spine, the cut-point of < or = 0 may be more appropriate. PMID- 15825703 TI - Evaluation of electrophysiological response to thymectomy in patients with myasthenia gravis. AB - Clinical response to thymectomy in patients with myasthenia gravis varies in terms of timing and degree of improvement. Electrophysiological test is a reasonably reliable and objective way to evaluate neuromuscular transmission in these patients. The purpose of this study was to evaluate the electrophysiological response before and after thymectomy by repetitive nerve stimulation test in order to estimate the timing of initial improvement, progress and degree of improvement after thymectomy. According to electrophysiological findings, it appeared that neuromuscular transmission in these patients might have begun to improve even as early as one week after thymectomy and steadily improved by the end of one year of the study corresponding to clinical improvement. PMID- 15825702 TI - Efficacy and safety of piribedil in early combination with L-dopa in the treatment of Parkinson's disease: a 6-month open study. AB - BACKGROUND: Piribedil is a non-ergot D2/D3 dopamine agonist with antagonistic effect on alpha2-adrenoceptors and lack of agonist properties at 5-HT2A/2C receptors. Previous studies indicated its efficacy in monotherapy as well as in combinatio' s disease in L-dopa-treated parkinsonian patients. PATIENTS AND METHOD: A 6-month, open-labeled, multicenter study was conducted in Thai Parkinsonian patients who were insufficiently controlled by L-dopa (< or = 600 mg/day). Piribedil 50 mg in retard form was titrated upward to 150 mg/day (50 mg tid) by the 5th week and up to 6 months as an add-on treatment. L-dopa daily dose was kept stable until the 3rd month and could be adjusted afterwards. The main efficacy parameter was the change in UPDRS part III score versus baseline over Full Analysis Set, score variation, and percentage of responders defined by at least 30% decrease from baseline of total UPDRS part III score. The secondary efficacy criteria were changes in L-dopa dose between the third month and the end of the study, UPDRS part II score variation, Hoehn and Yahr stage variation and Schwab and England Activities of Daily Living Scale variation. The acceptability of piribedil was assessed by physical examination, weight, blood pressure and heart rate as well as the reported adverse events. RESULTS: Twenty-nine patients (55.2% male) with the mean age of 64.0 +/- 7.2 years and mean duration of disease of 18.3 +/- 8.2 months were recruited The mean UPDRS part III score at baseline was 19.8 +/- 11.4. After 6-month treatment with piribedil, mean UPDRS part III score significantly decreased to 6.6 +/- 4.7 (p < 0.0001) with mean score variation of 13.3 +/- 10.3. Twenty-seven patients (93.1%) were responders. Mean UPDRS part II score was significantly decreased from 7.2 +/- 5.4 at baseline to 2.7 +/- 2.1 at the end of 6 months (p < 0.0001). Hoehn and Yahr stage and Schwab and England Activities of Daily Living Scale were also significantly improved Reported adverse events were mainly gastrointestinal symptoms. Blood pressure and heart rate were not significantly changed during the study period. Peak dose dyskinesia was reported only in one patient. Two patients (6.9%) were withdrawn because of adverse events. CONCLUSION: Piribedil was effective on motor symptoms during a 6-month treatment in early parkinsonian patients insufficiently controlled by L-dopa and it was well tolerated. PMID- 15825704 TI - Outcome after transsternal radical thymectomy for myasthenia gravis: 14-year review at Ratchaburi Hospital. AB - OBJECTIVE: To determine predictability factors, clinical features and outcome in the patients after transsternal radical thymectomy (TRT) for Myasthenia Gravis (MG). METHOD: A 14-year retrospective review (1990-2004) and analysis of medical data on 32 patients with MG who underwent TRT. Two patients were lost to follow up. Preoperative medication included anticholinesterase drugs and steroids. Plasmaphereses were used in 2 cases before operation. The modified Osserman clinical classification (Table 1) was used to define disease severity. The status of the patients was evaluated as follows: A (remission), B (improvement), C (no change), D (deterioration), E (death due to myasthenia gravis). Using univariate analysis, sex, age, onset, Osserman class and pathology correlated with outcome and statistical significance is defined as P < 0.05. RESULTS: Among the 30 patients in the present study with MG who underwent TRT women comprised 70% (21 of 30 patients), and mean age was 42.87 +/- 12.16 year (19-65 years). The preoperative duration of the disease ranged from 0.5 to 120 months (mean = 25.72 +/- 30.68 months). Clinical statuses of patients as assessed on initial evaluation were 13 patients (43.3%) were in class IIA, 6 (20%) in class IIB, and 11 (36.7%) in class III. (Table 2) The histology of thymus glands consisted of hyperplasia in 22 patients, normal in 2, atrophic thymus in 3, thymomas in 3, (2 were malignant thymomas). Hospital mortality was 0, but 9-month mortality was 3.33% (1 of 30 patients). The mean follow-up period was 41.80 +/- 53.89 months. Complete remission (A) was achieved in 40% of patients (12 of 30), and marked improvement (B) of MG in 40% (12 of 30), for a total benefit rate of 80%. 6.7% (2 of 30) were unchanged (C), 10% (3 of 30) were worse (D) and 3.3% died because of MG (E). Using univariate analysis on sex, clinical status by Osseman classification, and histopathology correlated significantly with outcome (P < 0. 05); 95.2% of women (20 of 21) benefited from the procedure, versus 44.4% of men (4 of 9). 100% (19 of 19) of patients in class IIA and IIB benefited from the procedure, versus 45.5% (4 of 9) of patients in class III. All patients with thymoma presented a less favorable outcome (deteriorated in 2, and died in 1) versus 11.1% (3 of 24) of patients without thymomas were deteriorated. CONCLUSION: The remission and improvement in 80% of patients suggested that adequate thymic tissue were removed with radical thymectomy. Female, absence of thymoma, thymic hyperplasia, patients in class IIA, and IIB (non-respiratory involvement) are favorable predictability prognostic factors. PMID- 15825705 TI - Knowledge and skill after brief ACLS training. AB - OBJECTIVES: To determine the knowledge level and skill base in nurse anesthetists before and after brief ACLS training, and again three months later. METHOD: Thirty nurse anesthetists were tested for knowledge and skill before ACLS training comprising 1-hr lecture and handout, and 1-hr simulation training. Concepts included ABCD, primary and secondary survey, management, medications, and algorithms for common problems. Skill practice comprised airway management, chest compression and practice with equipment. After the training, the nurse anesthetists were immediately tested and again three months later. RESULTS: Age of participants averaged 39.33 + or - 3.14 years and working experience 10.04 +/- 3.23 years. The knowledge and skill scores pre- vs post-training vs three-months later was 50.32 +/- 15.24 vs 75.40 +/- 10.29 (p < 0.001) vs 60.48 +/- 11.80 (p < 0.001) and 65.00 + 16.07 vs 79.67 +/- 10.80 (p < 0.001) vs 75.67 +/- 14.53 (p < 0.001), respectively. The pre-training vs three-months-post-training skill scores was not statistically different (p = 0.255). CONCLUSION: After the briefACLS training knowledge and skills were significantly improved, but knowledge was not retained at the post-training test levels until the 3-month check, albeit skills had persisted. More frequent ACLS education is necessary. PMID- 15825706 TI - Fit-and-fill analysis of trochanteric gamma nail for the Thai proximal femur: a virtual simulation study. AB - The present study present a three-dimensional virtual simulation method to evaluate the fit-and-fill effect of the insertion of a trochanteric gamma nail (TGN) in 98 Thai dadaveric proximal femora. The circular best fit of the 2 dimensional cross-section of the femoral canal and the nail at 4 levels [d100, d120, d140 and d160] which were located at 100, 120, 140 and 160 mm distal to the tip of the greater trochanter were calculated. The evaluation of each level included, 1) the diameters of the medullary canal, 2) the percentage of area filled by the nail in the unreamed medullary canal, 3) the minimal reamer diameter that required enlargement of the canal to accommodate TGN insertion, 4) the minimal inner cortical reaming thickness that needed to be removed, 5) the percentage of cortical bone area that needed to be removed prior to nail insertion and 6) the deviation of the nail center from the center of the medullary canal. The results showed that at 4 studied locations the diameter of unreamed medullary canal averaged 10.3 to 11.8 mm. The nail cross-section that could fill the medullary canal averaged 86.9-95.1%. The minimal reaming diameter for the medullary canal to accommodate the TGN insertion averaged 11.3 to 12.3 mm. The inner cortical thickness that should be removed averaged 0.6 to 0.8 mm. The cortical bone that needed to be removed averaged 13.6 to 19.3% of the total cortical area. The deviation of the nail center from the canal center averaged 0.3 to 0.8 mm. The present study showed some mismatching of the TGN to that of the Thai proximal femur. Appropriate reaming to prepare the medullary canal should be considered prior to TGN insertion to prevent technical problem. Future re-design of the implant may be considered for Thai patients. PMID- 15825707 TI - Prevalence of diabetic retinopathy in relation to duration of diabetes mellitus in community hospitals of Lampang. AB - Three thousand and forty nine patients diagnosed with diabetes mellitus were examined in 13 community (district) hospitals in Lampang from January to December 2002. Complete eye examination with standard protocol was used to determine the prevalence and severity of diabetic retinopathy. The prevalence of the background or non-proliferative diabetic retinopathy (BDR or NPDR) was 18.9% and proliferative diabetic retinopathy (PDR) was 3% in all age groups. For the relationship of the duration of diabetes, it showed that the longer the duration of diabetes the higher the prevalence of diabetic retinopathy. In BDR or NPDR, the retinopathy varied from 13.11 to 22.91% in persons having diabetes for less than 10 years and up to 42.86% in those with diabetes for up to 20 years. In the PDR group, the prevalence was 2.15 to 2.42% in persons with diabetes for less than 10 years and up to 10.20% for those with diabetes for up to 20 years. The severity of retinopathy was found to be not only related to a longer duration of diabetes but also related to higher glycosylated hemoglobin levels, higher systolic blood pressure and the presence of proteinuria. PMID- 15825708 TI - Prevalence of CagA and VacA genotype of Helicobacter pylori in Thai children. AB - Seroprevalence of Helicobacter pylori infection in children is variable according to geographical location and family sanitation. A previous study in Bangkok showed an incidence of 25.5% in 1998. The higher incidence in the urban and rural area is predicted in lower economic classes and poor sanitation. OBJECTIVE: To study the occurance of CagA and VacA genotype in Thai children using the Western blot technique. MATERIAL AND METHOD: Sera of 159 Thai native children aged 0-15 year without associated abdominal pain from different provinces in 4 parts of the Kingdom of Thailand were tested with the rapid screening test for H. pylori. The positive specimen was further tested with the Western blot technique for determination of Urea A (p37), CagA (p116) and VacA (p89). RESULT: Fiftyfive andfifty two (34.6%) were tested positive by the rapid test while 32.7% were positivefor the band of current infection marker (CIM). The 28 selected positive sera with complete history of housing and water supply were analysed. Thai children living in urban areas have a higher prevalence and the CagA+, VacA+ are found in 96.43% of infected patients. The transmission may be through the water supply. CONCLUSION: A high prevalence of Helicobacter pylori infection was found in childhood period in urban areas and may be associated with the local water supply. PMID- 15825709 TI - Aneurysmal third nerve palsy. AB - To study the clinical characteristics, treatment and outcome of patients with aneurysmal third nerve palsy. Eleven patients with isolated third nerve palsy from an intracranial aneurysm from 1998 to 2002 at Ramathibodi Hospital were reviewed retrospectively. The average age was 60 years. Ipsilateral headache and/or retroorbital pain occured in 80 percent of the patients. Ten patients had unilateral disease, whereas one patient had bilateral involvement. The most common site of aneurysm was at the origin of the posterior communicating artery. Endovascular treatment with coil embolization provided successful occlusion as well as neurosurgical clipping. Recovery of third nerve function was found in all patients but had variable degrees. Patients who received early treatment, especially within 10 days after onset of oculomotor dysfunction appeared to have the best chance of recovery. Aberrant regeneration developed in 5 cases. The commonest sign was lid retraction during adduction and downward gaze. PMID- 15825710 TI - Management of blunt duodenal injuries. AB - BACKGROUND/AIMS: Blunt duodenal injuries are relatively rare. Diagnosis is usually delayed resulting in significant morbidity and mortality. Treatment of the injured duodenum varies according to severity of injury and duration before diagnosis. Duodenal fistulas are serious complications with potential mortality. The purpose of this study was to examine results of treatment of patients with blunt duodenal injuries at our institution. PATIENTS AND METHOD: This is a retrospective study of patients who sustained blunt duodenal injuries and were admitted to King Chulalongkorn Memorial Hospital, Bangkok, Thailand from January 1990 to December 2003. During the study period, management of duodenal injuries at our institution depended largely on severity of injuries, timing of diagnosis and the presence of retroperitoneal infections. Intramural hematoma of the duodenum was treated conservatively. Uncomplicated wounds of the duodenum (grade II and grade III injury) with no obvious retroperitoneal infections were treated by simple duodenal repair. Pyloric exclusion was performed in cases of difficult duodenal repair and/or delayed diagnosis (> 24 hours after injury) with obvious evidence of retroperitoneal infections. RESULTS: Twenty six patients were entered into the study. Five patients (19.2%) had intramural hematoma of the duodenum, all were successfully treated by conservative treatment. Twenty one patients (80.8%) had transmural tear of the duodenal wall. Ten of them (47.6%) underwent simple repair, 10 (47.6%) underwent simple repair of the duodenal wounds combined with pyloric exclusion (2 of them underwent the operations elsewhere), and 1 (4.8%) underwent pancreaticoduodenectomy. Seven patients who had transmural tear of the duodenum developed complications (33.3%). Two patients had duodenal fistulas (9.5%); 1 in the simple repair group and 1 in the pyloric exclusion group. One patient who underwent pyloric exclusion had leakage of the gastrojejunostomy anastomosis with intact duodenal repair resulting in a complicated and prolonged hospital course. There was no mortality in this study. CONCLUSIONS: The outcome in management of blunt duodenal injuries at our institution was acceptable with low morbidity and no mortality. Intramural hematomas were safely treated conservatively. Uncomplicated duodenal wounds were treated by simple suture repair. Pyloric exclusion was a useful additional procedure in patients with complicated duodenal injuries. Retroperitoneal infections was a strong indication to perform this procedure in addition to simple repair of the duodenal wounds. Pancreaticoduodenectomy should be reserved for only severed combined duodenal and pancreatic head injuries. PMID- 15825711 TI - Comparison of effectiveness between gas flow 1 and 2 l.min(-1) for general anesthesia in infants and children. AB - OBJECTIVES: To determine 1) Success rate of using fresh gas flow (FGF) 1 l.min( 1) compared to 2 l.min(-1) in pediatric patients 2) Necessity of using anesthetic agent analyzer 3) predicting volatile anesthetic concentration 4) cost difference. METHOD: Seventy-seven patients (age 10 days to 8 years) who underwent general anesthesia were randomly allocated into 2 groups: the control group (FGF 2 l.min(-1)) and the study group (FGF 1 l.min(-1)). Outcome measures included system leakage, SaO2, PECO2, FiO2, Fi and FeN2O, isoflurane dial setting, Fi and Fe isoflurane, isoflurane mass consumed and sodalime used. RESULTS: There was no difference between the groups regarding demographic data, duration of surgery and anesthesia. Success rate in using FGF 2 l.min(-1) was 100% and FGF 1 l.min(-1) was 92%. All failure cases (8%) were due to system leakage. The necessity of using a capnometer was similar at 5.3-7.7% in both groups. FiO2 was > or = 0.3 at any time. FiN2O and FeN2O were not different. Fi isoflurane was 13-15% lower than dial setting. Overall savings from using FGF 1 l/min was 37.8%. CONCLUSION: FGF 1 l.min(-1) could be safely used in most pediatric patients with lower cost. Capnometer was recommended, whereas FiO2 and Fi isoflurane could be clinically adjusted PMID- 15825712 TI - Prevalence of hepatitis B virus and hepatitis C virus co-infection with human immunodeficiency virus in Thai patients: a tertiary-care-based study. AB - BACKGROUND: Hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV share the route of transmission. HBV or HCV co-infection with HIV has been associated with a reduced survival rate, an increased risk of progression to severe liver disease, and an increased risk of hepatotoxicity associated with active antiretroviral therapy. Information regarding prevalence of HBV and HCV co infection with HIV in Thailand is limited. PATIENTS AND METHOD: A cross-sectional study of prevalence and risk factors of HBV and HCV co-infection in HIV-infected patients was conducted. All HIV-infected patients who were cared for in March 2003 at Ramathibodi Hospital were included. RESULTS: There were 529 HIV-infected patients with a mean age of 36.7 years and 56.5% males. Of these, 58.8% lived in Bangkok, whereas, the others were from provincial areas. Heterosexual contact were the acquisition of HIV infection in 98.1% of all patients. The prevalence of HBV infection was 8.7%, and HCV infection was 7.8%. There was no difference between the prevalence of these infections in Bangkok and provincial areas (p = 0.115). History of intravenous drug use was associated with both HBV and HCV co infection (p < 0.001). HCV co-infection group was also associated with male gender (p = 0.002) and elevated serum alanine transaminase (ALT) level (p = 0.0003). CONCLUSIONS: The prevalence of HBV and HCV co-infection with HIV in Thai patients is significant. In the author s resources-limited setting, history of intravenous drug use is a major indicator to screen for both HBV and HCV co infection. Male gender and elevated serum ALT level are also suggestive of HCV co infection. PMID- 15825713 TI - Retrospective study of pathologically proven colitis in King Chulalongkorn Memorial Hospital. AB - To assess the basic clinicopathological information of colonic inflammation in Thai patients, the authors retrospectively analyzed the 249 biopsied cases with pathological diagnosis as colitis in King Chulalongkorn Memorial Hospital during the five-year period from 1998 to 2002. All subjects were included in this study, whether newly diagnosed or follow-up cases. There were 122 (49%) males and 127 (51%) females with the mean age of 51 years. Non-specific colitis was the most frequent histological diagnosis (72%), followed by infectious colitis (12%), in particular mycobacterial infection. The biopsy specimens were commonly obtained from the rectosigmoid colon (38%). Mucous bloody diarrhea (28%), watery diarrhea (26%), and lower gastrointestinal bleeding (19%) were the three most common symptoms at presentation in order of frequency. PMID- 15825714 TI - Comparison of different culture methods on bacterial recovery in hemodialysis fluids. AB - To examine the culture method that could provide the highest bacterial recovery, 143 reverse osmosis water samples used in hemodialysis were collected for comparison of the media (Tryptic Soy Agar, TSA vs Reasoner's 2A Agar, R2A), the temperature (20 degrees C vs 37 degrees C), the duration of incubation (48-hour vs 7-day), and the culture technique (membrane filtration vs spread plate methods). The European Best Practice Guideline method, R2A at 20 degrees Cfor 7 day incubation provided higher bacterial recovery than the Association for the Advancement of Medical Instrumentation (AAMI) method, TSA at 37 degrees C for 48 hour incubation. The membrane filtration method gave better yield than the spread plate method. As such, the European Best Practice Guideline method in combination with the membrane filtration technique would be the culture method of choice for hemodialysis fluids. PMID- 15825715 TI - Determination of the thecal sac ending using magnetic resonance imaging: clinical applications in craniospinal irradiation. AB - PURPOSE: To determine level of the thecal sac ending in unaffected individuals of young age-group and in patients with cerebrospinal fluid (CSF) seeding tumor using Magnetic Resonance Imaging (MRI). MATERIAL AND METHOD: MRI of the lumbosacral spine of 253 cases (age range 1-40 years) had been reviewed. Of these, 20 cases were known having CSF seeding tumors. Two hundred and thirty three cases of unaffected individuals were used as control group. Sagittal MR images were used to determine the level of thecal sac ending by referencing on the corresponding vertebral body and intervertebral disc. RESULTS: The range of the thecal sac ending level in unaffected population varied from the lower one third of L5 to the middle one-third of S3. About 97% of cases, the distal end of thecal sac terminated at the S2-3 intervertebral disc space or higher. In patients with CSF seeding tumor, the range of thecal sac ending was from the middle one-third of L5 to the S2-3 intervertebral disc space. There was no statistically significant difference of thecal ending level between unaffected group and patients with CSF seeding tumors (p=0.19). CONCLUSION: Placing inferior border of radiation field at the middle one-third of S3 vertebra would help to cover the entire subarachnoid space in all cases. Without MR machine, this study may be used as a general guideline for placing the inferior border for spinal irradiation. PMID- 15825716 TI - Four years follow-up of bone mineral density change in premenopausal women with systemic lupus erythematosus. AB - OBJECTIVE: To measure the change in bone mineral density (BMD) in premenopausal women with systemic lupus erythematosus (SLE) during 4 years of follow-up, and to identify the role of glucocorticoid and disease related variables. METHOD: Premenopausal women with SLE were clinically evaluated and underwent BMD measurement of the lumbar spine, femoral neck and trochanter by dual energy x-ray absorptiometry. RESULTS: 106 SLE patients were evaluated with a mean age of 31.7 +/- 7.5 years, duration of SLE 2.5 +/- 2.6 years, mean daily dose 17.1 +/- 14 mg/d, duration of prednisolone treatment 16.3 +/- 19.9 months during 4 years of follow-up. There was no significant change in BMD at the lumbar spine (1.051 +/- 0.15 vs 1.052 +/- 0.14 vs 1.056 +/- 0.17 vs 1.056 +/- 0.19; p = 0.27), femoral neck (0.861 +/- 0.12 vs 0.867 +/- 0.12 vs 0.846 +/- 0.12 vs 0.844 +/- 0.12, p = 0.28) and trochanter (0.718 +/- 0.12 vs 0.726 +/- 0.13 vs 0.717 +/- 0.13 vs 0.709 +/- 0.14; p = 0.26) at the baseline, first, second and fourth year follow-up study. Furthermore, annual percentage BMD changes were not significant in lumbar BMD (p = 0.37), femoral neck BMD (p = 0.65) and trochanteric BMD (p = 0.47) during the 4 years follow-up study. The average annual percentage change of BMD was not significantly associated with change in age, body mass index (BMI), disease activity, disease severity, disease duration and prednisolone treatment. In addition, there were no significant bone changes between subgroups treated with < or = 7.5 mg and > 7.5 mg daily dose of prednisolone as indicated by BMD at the lumbar spine, femoral neck and trochanter as well as annual percentage BMD changes over the study period. CONCLUSION: There was no significant change of lumbar spine, femoral neck or trochanteric BMD in premenopausal SLE women treated with corticosteroid. These findings suggest that low dose prednisolone may not be detrimental to bone in premenopausal women with SLE during longterm treatment. PMID- 15825717 TI - Randomized double-blind study of phenylephrine 2.5% vs 10% on pupillary dilation. AB - PURPOSE: To compare the efficacy of phenylephrine 2.5% versus 10% on pupillary dilation, and also compare their side-effects. METHOD: Patients at the Eye Clinic Srinagarind Hospital were randomized into two groups. Patients in group 1 received 1% tropicamide and 10% phenylephrine, whereas those in group 2 received 1% tropicamide and 2.5% phenylephrine. Pupil diameter, blood pressure and heart rate were measured before and after eyedrop instillation. RESULTS: Five hundred and sixty four patients were randomized into 293 patients (group 1) and 271 patients (group 2), using simple random sampling method. Mean pupil diameters before instillation in group 1 were 4.43 +/- 1.13 mm in the right eye and 4.31 +/ 0.95 mm in the left eye, whereas those in group 2 were 4.45 +/- 1.0 mm in the right eye and 4.32 +/- 0.92 mm in the left eye. After the instillation, the mean pupil diameters in group 1 were 7.58 +/- 0.96 mm in the right eye and 7.60 +/- 1.03 mm in the left eye, whereas those in group 2 were 7.17 +/- 1.04 mm in the right eye and 7.07 +/- 1.06 mm in the left eye. The difference was statistically significant (P < 0.05). There was no significant difference in mean systolic and diastolic blood pressure after instillation between the two groups. However, the mean heart rate after instillation in group 1 was greater than the value in group 2 with statistically significant difference. CONCLUSION: Pupillary dilation with 1% tropicamide and 10% phenylephrine is more effective than 1% tropicamide and 2.5% phenylephrine with statistically significant difference. After single dose instillation, there was no significant difference in the mean blood pressure between the two groups. PMID- 15825718 TI - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS): a case report in Thailand. AB - Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a rare congenital disease with high mortality rate. The authors report a case of a female term infant with massive abdominal distension at birth. Abdominal ultrasonography revealed a huge cystic mass resolved after urinary catheterization. Exploratory laparotomy was performed and intraoperative findings were an enlarged urinary bladder, microcolon, short bowel and malrotation of the small intestine. Ladd's procedure, ileostomy and vesicostomy were performed. Pathological findings of rectal biopsy revealed normal ganglion cells. Although prokinetic drugs were given for promoting bowel motility, enteral feedings were not tolerated. She died from septicemia at the age of 50 days. PMID- 15825719 TI - The rabies epidemic on Flores Island, Indonesia (1998-2003). AB - Flores is an isolated previously rabies-free Indonesian island which has been experiencing a canine rabies outbreak which resulted in at least 113 human deaths. It started with the importation of three dogs from rabies endemic Sulawesi in September of 1997. Local authorities responded with massive killing of dogs starting in early 1998. Approximately 70% of the dogs, in the district where rabies had been introduced, were killed during that year, yet canine rabies still exists on Flores at this time (June 2004). Approximately 50 percent of dog bite cases and human deaths were in children under 15 years of age. Between 1998 and the end of 2002, 3,389 post-exposure rabies treatments were provided and none of the treated subjects died. Almost all patients were given the Thai Red Cross intradermal rabies postexposure treatment regimen using tissue culture rabies vaccine. A small but unknown number of patients with very severe exposures also received equine rabies immune globulin. This occurrence indicates that even massive culling of the dog population, without an intensive vaccination campaign of the survivors, will not arrest an outbreak even if it occurs on a small island. PMID- 15825720 TI - Electrophysiological concept of ventricular defibrillation mechanism. AB - Sudden cardiac death is a major health problem in most industrialized countries around the world including Thailand. It is mainly caused by ventricular fibrillation (VF). Currently, defibrillation is the only effective clinical treatment of this fatal arrhythmia. Although defibrillation mechanism has been investigated extensively for many decades, its definite mechanism is still debated. It is known that understanding the basic mechanism of defibrillation is essential to develop better treatment of VF: In the present article, seven hypotheses commonly proposed as the mechanism of ventricular defibrillation are reviewed. Since research in the field of defibrillation mechanism is dynamic, the present review is to update the information to clinicians and basic investigators on the mechanism of defibrillation available to date. PMID- 15825721 TI - Sex reassignment surgery in Thailand. AB - Many years ago Thai society considered transsexualism (Gender identity disorder or Gender dysphoria) which is commonly known as Kathoey (a word originally used to denote hermaphrodites), Sao Prapet Song or Tut (as in 'Tootsie') were low class citizens, dirty dressing and had to hide in a dark corner selling their services as prostitutes. This made us unwilling to do sex reassignment surgery for this group of people because the idea of eradicating normal sexual organs for the purpose that was not accepted by the society. Consequently the authors have experience in cases where these people wandered seeking doctors who had no competency nor enough experience to do the surgery. The authors could not inhibit the desire of these people who usually suffer from gender identity disorder from strongly wishing to change their genital sex to the sex they want. The outcome of the surgery was not satisfactory for the patients. There were complications and sequelae which caused the authors to correct them later which might be more difficult than doing the original surgery. In addition there were more studies about the etiology and affect of the disorder on these people that changed the social point of view. The women who wanted to be a him and men who would like to be a her should be considered as patients who need to be cured to set the harmony about their genetic sex and the desire to be the opposite sex and also to be regarded by others as a member of that other sex. The treatments of transsexualism usually begin with conventional psychiatric and endocrinological treatment to adjust the mind to the body. For those who failed conservative treatment in adjusting the mind to the body then sex reassignment surgery will be the only way to transform their body to their mind and give the best result in properly selected patients. Preecha Tiewtranon, the pioneer in sex reassignment surgery in Thailand, did his transsexualism case in 1975 together with Dr. Prakob Thongpeaw. Sex reassignment surgery has been taught in Chulalongkorn University Hospital since 1983. (At present, it is the only medical school in Thailand that has sex reassignment surgery systematically taught and with good results). There have been many versions of development of the surgical techniques to gain better and better results. PMID- 15825722 TI - Riddell named to lead AABP. PMID- 15825723 TI - Veterinarian part of US-Russian collaboration on anthrax sensor. PMID- 15825724 TI - Diagnostic tests for mucopolysaccharidosis. PMID- 15825725 TI - More on vegetarian/vegan cat foods. PMID- 15825726 TI - Endorses Heifer International's tsunami relief. PMID- 15825727 TI - An alternative method to assess efficiency of estrus detection in dairy herds. PMID- 15825729 TI - What is your diagnosis? Pneumocephalus. PMID- 15825728 TI - What is your diagnosis? A wood fragment was detected within the mediastinum, penetrating the cranial aspect of the pericardial sac and in contact with the right auricle. PMID- 15825730 TI - Future needs and recommendations for leadership in veterinary medicine. PMID- 15825731 TI - Response of a veterinary college to career development needs identified in the KPMG LLP study and the executive summary of the Brakke study: a combined MBA/DVM program, business certificate program, and curricular modifications. AB - In the present market, veterinarians with a strong background in career development, practice management, and business skills have a clear advantage in achieving financial success. Although there is ample evidence that the scientific and clinical skills of veterinary college graduates are high, there are also data that suggest that additional capabilities in the business realm may promote greater economic success. As noted in the KPMG executive summary, the field of veterinary medicine must make changes in its "current business practices and attitudes" to be successful in the future. Furthermore, the KPMG study found that 36% of industry employers reported that some jobs within their companies had specific job requirements that were not met by a veterinarian with only a veterinary medical degree. The areas of additional training most often cited included business, administration, personnel management, sales and marketing, and financial skills. Yet, Lewis and Klausner found that veterinarians reported challenges in the business realm, such as "how business works and how business goals are translated into action. This challenge held true for veterinarians in industry, academia, government, and private practice." The present gender trends in the field of veterinary medicine provide additional impetus to make career development and business skills training more prevalent. Presently, women comprise >65% of the veterinary student population and approximately 45% of all practicing veterinarians. In some areas of practice, the rate is much higher. For example, in 2002, women comprised 48.2% of all small animal exclusive private practitioners. Unfortunately, the KPMG study found that female veterinarians in private practice report lower self-evaluation of business management and financial skills, compared with their male cohorts. Female veterinarians in nonprivate practice report lower self-evaluation in communication, personnel management, business management, and marketing skills than that reported by males. As a result of these pressing needs, CSU CVMBS has undertaken a major initiative to improve the veterinary practice management and business skills training of veterinary students by offering a variety of options to gain this knowledge: a combined MBA/DVM degree program, a Business Certificate Program for Health Professions, and core curriculum courses. In this way, students can select the amount of focus they want to place on career development and business skills as they earn their DVM degree, to best ensure that they become successful veterinarians. PMID- 15825732 TI - The role of glucosamine and chondroitin sulfate in treatment for and prevention of osteoarthritis in animals. PMID- 15825733 TI - Evaluation of diphenhydramine as a sedative for dogs. AB - OBJECTIVE: To determine and compare levels of sedation achieved by IM administration of diphenhydramine, saline (0.9% NaCl) solution, and acepromazine in dogs. DESIGN: Prospective randomized study. ANIMALS: 56 dogs. PROCEDURE: Dogs were randomly assigned to receive diphenhydramine at 2, 4, or 8 mg/kg (0.9, 1.8, or 3.6 mg/lb, respectively) i.m.; acepromazine at 0.1 mg/kg (0.05 mg/lb) i.m.; or saline solution at 0.05 mL/kg (0.02 mL/lb) i.m. Sedation was assessed by use of a 6-category descriptive system based on observation and interaction. RESULTS: Dogs in the acepromazine group had significantly higher sedation scores than did dogs in the saline solution or diphenhydramine groups at 30 minutes. Dogs in the diphenhydramine groups did not have significantly different sedation scores from dogs in the saline solution group at any time point. CONCLUSIONS AND CLINICAL RELEVANCE: Diphenhydramine did not cause clinically appreciable sedation in healthy dogs. Diphenhydramine is not suitable as a sole sedative prior to general anesthesia in dogs. PMID- 15825734 TI - Use of a particulate extracellular matrix bioscaffold for treatment of acquired urinary incontinence in dogs. AB - OBJECTIVE: To evaluate use of a particulate bioscaffold consisting of the extracellular matrix (ECM) of the urinary bladder from pigs for treatment of acquired urinary incontinence in dogs resistant to medical treatment. DESIGN: Case series. ANIMALS: 9 female dogs with acquired urinary incontinence. PROCEDURE: In 6 dogs, 30 mg of particulate ECM in 1.0 mL of a carrier consisting of glycerin and saline 10.9% NaCI) solution was injected into each of 3 equally spaced sites around the circumference of the internal urethral sphincter via an endoscopic technique. In the remaining 3 dogs (control dogs), 1.0 mL of the carrier alone was injected in 3 equally spaced sites around the circumference of the internal urethral sphincter in a similar manner. RESULTS: For dogs treated with the ECM, median duration of urinary continence following treatment was 168 days (range, 84 to 616 days), whereas for the control dogs, median duration of urinary continence following the procedure was 14 days (range, 7 to 31 days). Two of the 3 control dogs were treated with the ECM at the end of the study and were continent for 119 and 252 days. No adverse effects were observed in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that endoscopically guided injection of particulate ECM into the internal urethral sphincter may be useful for the treatment of acquired urinary incontinence in female dogs. PMID- 15825735 TI - Minimum alveolar concentration of isoflurane in mechanically ventilated Dumeril monitors. AB - OBJECTIVE: To determine minimum alveolar concentration (MAC) of isoflurane in mechanically ventilated Dumeril monitors (Varanus dumerili). DESIGN: Prospective study. ANIMALS: 10 healthy adult Dumeril monitors. PROCEDURE: Anesthesia was induced with isoflurane in oxygen delivered through a face mask. Monitors were endotracheally intubated, and end-tidal and inspired isoflurane concentrations were continuously measured. After equilibration at an end-tidal-to-inspired isoflurane concentration ratio of >0.9 for 20 minutes, an electrical stimulus (50 Hz, 50 V) was delivered to the ventral aspect of the tail for up to 1 minute and the monitor was observed for purposeful movement. End-tidal isoflurane concentration was then decreased by 10%, and equilibration and stimulation were repeated. The MAC was calculated as the mean of the lowest end-tidal isoflurane concentration that prevented positive response and the highest concentration that allowed response. A blood sample for blood gas analysis was collected from the tail vein at the beginning and end of the anesthetic period. RESULTS: Mean +/- SD MAC of isoflurane was 1.54 +/- 0.17%. Mean heart rates at the upper and lower MAC values were 32.4 +/- 3 beats/min and 34 +/- 4.5 beats/min, respectively. During the experiment, PaCo2 decreased significantly from 43.1 mm Hg to 279 mm Hg and blood pH and HCO3 concentration increased significantly from 7.33 to 7.64 and from 25.3 to 32.9 mmol/L, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The MAC of isoflurane in Dumeril monitors was similar to that reported in mammals but lower than values reported in other reptiles. This difference may be reflective of the more advanced cardiovascular physiologic features of monitor lizards. PMID- 15825736 TI - Clinical, echocardiographic, and electrocardiographic abnormalities in Boxers with cardiomyopathy and left ventricular systolic dysfunction: 48 cases (1985 2003). AB - OBJECTIVE: To identify clinical, echocardiographic, and electrocardiographic abnormalities in Boxers with cardiomyopathy and echocardiographic evidence of left ventricular systolic dysfunction. DESIGN: Retrospective study. ANIMALS: 48 mature Boxers. PROCEDURE: Medical records were reviewed for information on age; sex; physical examination findings; and results of electrocardiography, 24-hour ambulatory electrocardiography, thoracic radiography, and echocardiography. RESULTS: Mean age of the dogs was 6 years (range, 1 to 11 years). Twenty (42%) dogs had a systolic murmur, and 9 (19%) had ascites. Congestive heart failure was diagnosed in 24 (50%) dogs. Seventeen (35%) dogs had a history of syncope. Mean fractional shortening was 14.4% (range, 1% to 23%). Mean left ventricular systolic and diastolic diameters were 4.5 cm (range, 3 to 6.3 cm) and 5.3 cm (range, 3.9 to 7.4 cm), respectively. Twenty-eight (58%) dogs had a sinus rhythm with ventricular premature complexes (VPCs), and 20 had supraventricular arrhythmias (15 with atrial fibrillation and 5 with sinus rhythm and atrial premature complexes). Sixteen of the dogs with supraventricular arrhythmias also had occasional VPCs. Morphology of the VPCs seen on lead II ECGs was consistent with left bundle branch block in 25 dogs, right bundle branch block in 8, and both in 11. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that Boxers with cardiomyopathy and left ventricular dysfunction frequently have arrhythmias of supraventricular or ventricular origin. Whether ventricular dysfunction was preceded by electrical disturbances could not be determined from these data, and the natural history of myocardial disease in Boxers requires further study. PMID- 15825737 TI - Bilateral cavernous sinus syndrome in dogs: 6 cases (1999-2004). AB - OBJECTIVE: To determine clinical features, diagnostic imaging abnormalities, underlying disease, disease progression, and outcome in dogs with bilateral cavernous sinus syndrome. DESIGN: Retrospective study. ANIMALS: 6 dogs. PROCEDURE: Dogs were included if clinical signs consistent with bilateral cavernous sinus syndrome (i.e., deficits of the third, fourth, and sixth cranial nerves and at least 1 of the first 2 branches of the fifth cranial nerve) were present and a lesion of the cavernous sinus was identified by means of diagnostic imaging or postmortem examination. RESULTS: 5 dogs were evaluated because of problems referable to abnormal ocular motility or pupillomotor dysfunction, and 1 dog was evaluated because of partial motor seizures involving the face and bilateral mydriasis. Four dogs had neurologic signs referable to an extrasinusoidal lesion at the time of initial examination, and the remaining 2 dogs eventually developed extrasinusoidal signs. Besides neuroanatomic location, the only consistent neuroimaging feature was variably intense, heterogeneous enhancement of cavernous sinus lesions. Neoplasia was histologically confirmed as the underlying cause in 5 of the dogs and was suspected in the remaining dog. Median survival time for the 4 dogs that were treated was 199 days (range, 16 to 392 days). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that bilateral cavernous sinus syndrome is rare in dogs but should be suspected in dogs with compatible clinical signs. Affected dogs have a poor prognosis, and dogs with clinical signs of bilateral cavernous sinus syndrome should be systematically evaluated for neoplastic disease. PMID- 15825738 TI - Evaluation of factors that affect embryonic loss in dairy cattle. AB - OBJECTIVE: To identify potential risk factors for embryonic loss before 35 to 42 days of gestation in dairy cattle. DESIGN: Prospective observational study. ANIMALS: 381 cows. PROCEDURE: Body condition score was determined at the time of artificial insemination (AI; day 0) and on days 20, 23, and 27 and between days 35 and 41; serum progesterone concentration was measured on days 0; 20 or 21; and 23, 24, or 25. Cows were excluded from analyses if day 0 serum progesterone concentration was > or = 1.0 ng/mL and classified as pregnant on day 23 if serum progesterone was > 1.5 ng/mL on day 20 or 21 and day 23, 24, or 25. Cows were examined via transrectal ultrasonography on day 27 or 28 and rectally palpated for pregnancy on days 35 to 41. RESULTS: 39% of cows that were pregnant on day 23 lost their embryo by day 27, and 18% of cows that were pregnant on day 27 or 28 were not pregnant on days 35 to 41. Breeding a pregnant cow posed the greatest risk for embryonic loss at both time periods. Mean serum progesterone concentrations on day 21 or 22 and day 23, 24, or 25 were lowest for cows that lost an embryo between days 24 and 28. Cows with a linear somatic cell count score > 4.5 before AI were twice as likely to lose the embryo by 35 to 41 days, compared with cows with a score < 4.5. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that embryonic loss could be reduced by more accurate estrus detection, reducing mastitis, and strategies to improve progesterone concentration after breeding. PMID- 15825739 TI - Milk flow and udder health in cows after treatment of covered teat injuries via theloresectoscopy: 52 cases (2000-2002). AB - OBJECTIVE: To determine milk flow, somatic cell counts (SCCs), and the incidence of clinical mastitis in cows that had undergone theloresectoscopy for treatment of teat stenosis caused by mucosal detachment in the region of the streak canal or Furstenberg's rosette. DESIGN: Retrospective study. ANIMALS: 52 cows with teat stenosis that were treated via theloresectoscopy. PROCEDURE: Medical records of eligible cows were reviewed. Additional data regarding milking ease, SCC, development of clinical mastitis of the affected gland, and whether the cow remained in the herd were collected via owner-completed questionnaires. RESULTS: 49 of 52 questionnaires were completed. At referral, teat sinusitis was diagnosed in 29 of 52 cows. Milk flow was normal in 38 of 41 treated teats at discharge and in 24 of 40 during the next lactation. Thirteen of 49 cows were culled during the next lactation because of abnormal udder health. High SCC and teat sinusitis at referral and development of clinical mastitis during the 10-day period after surgery resulted in high SCCs in the remainder of the current lactation. The incidence of clinical mastitis during the remainder of the current and during the next lactation was higher in cows that had teat sinusitis at the time of surgery, compared with those that did not. CONCLUSIONS AND CLINICAL RELEVANCE: Teat stenoses resulting from mucosal lesions in the region of the streak canal or Furstenberg's rosette may be successfully treated via theloresectoscopy. Inflammation of the teat sinus and gland at the time of surgery may adversely affect udder health. PMID- 15825740 TI - Optical coherence tomography in uncooperative children under general anesthesia. PMID- 15825741 TI - What's your diagnosis? Congenital iris ectropion syndrome associated with glaucoma. PMID- 15825742 TI - A shining light in the Land of the Midnight Sun. PMID- 15825743 TI - The clinical spectrum and treatment outcome of retinoblastoma in Indian children. AB - PURPOSE: To study the clinical spectrum and treatment outcome of retinoblastoma in Indian children. PATIENTS AND METHODS: This retrospective study analyzed 488 eyes of 355 retinoblastoma patients treated at a tertiary care ophthalmic hospital in southern India during a 14-year period. RESULTS: Retinoblastoma involved one eye in 177 (50%) and both eyes in 178 (50%) patients. Mean age at presentation was 23.98 +/- 23.37. PMID- 15825744 TI - A systematic review of the applicability and efficacy of eye exercises. AB - PURPOSE: To examine the current scientific evidence base regarding the efficacy of eye exercises as used in optometric vision therapy. METHODS: A search was performed of the following databases: Allied and Complementary Medicine Database, Cochrane Database of Systematic Reviews, Cochrane Register of Controlled Trials, EMBASE, and MEDLINE. Relevant articles were reviewed and analyzed for strengths and weaknesses. Pertinent sections of classic texts were studied to provide a historical basis and to serve as a source for additional early references. RESULTS: Forty-three refereed studies were obtained. Of these, 14 were clinical trials (10 controlled studies), 18 review articles, 2 historical articles, 1 case report, 6 editorials or letters, and 2 position statements from professional colleges. Many of the references listed by the larger reviews were unpublished or published in obscure or nonrefereed sources and therefore were not accessible. CONCLUSIONS: Eye exercises have been purported to improve a wide range of conditions including vergence problems, ocular motility disorders, accommodative dysfunction, amblyopia, learning disabilities, dyslexia, asthenopia, myopia, motion sickness, sports performance, stereopsis, visual field defects, visual acuity, and general well-being. Small controlled trials and a large number of cases support the treatment of convergence insufficiency. Less robust, but believable, evidence indicates visual training may be useful in developing fine stereoscopic skills and improving visual field remnants after brain damage. As yet there is no clear scientific evidence published in the mainstream literature supporting the use of eye exercises in the remainder of the areas reviewed, and their use therefore remains controversial. PMID- 15825745 TI - Comparison of eccentric fixation measurements using the streak target of an ophthalmoscope and a traditional visuoscopy target. AB - PURPOSE: To compare measurements of eccentric fixation using two different targets for fixation, a traditional visuoscope target and the streak target of an ophthalmoscope. PATIENTS AND METHODS: Monocular fixation was evaluated using visuoscopy in 47 patients ranging in age from 4 months to 22 years. Visuoscopy measurements were compared using both the traditional visuoscope target and the streak target of a Welch Allyn ophthalmoscope. The streak light was rotated both horizontally and vertically to detect both horizontal and vertical eccentric fixation. RESULTS: The streak target improved testability of eccentric fixation in children younger than 3 years compared to the traditional visuoscope target (75% versus 30%). All of the patients older than 3 years were testable using both targets, with both methods yielding the same results. All of the patients with a visual acuity of at least 20/20 also demonstrated central fixation using both techniques. CONCLUSIONS: Using the streak of a Welch Allyn ophthalmoscope as a visuoscope target allows for testing of eccentric fixation in children younger than 3 years. PMID- 15825746 TI - Long-term results of trabeculectomy with different concentrations of mitomycin C in refractory developmental glaucoma. AB - PURPOSE: To evaluate the long-term results of trabeculectomy with two different concentrations of mitomycin C in refractory developmental glaucoma. PATIENTS AND METHODS: A total of 48 eyes of 37 patients with previous failure of congenital glaucoma surgery underwent mitomycin C-augmented trabeculectomy. Data were analyzed retrospectively, and patients were divided into two groups. Group 1 consisted of 18 patients (25 eyes) who underwent trabeculectomy with a 0.4 mg/mL mitomycin C solution and group 2 consisted of 19 patients (23 eyes) who underwent trabeculectomy with a 0.2 mg/mL mitomycin C solution. Pre- and postoperative intraocular pressure (IOP), best-corrected visual acuity, bleb characteristics, success rate, and complications were compared for both groups. RESULTS: Mean time between primary surgery and mitomycin C-augmented trabeculectomy was 19.3 months (range, 11 months to 7 years). Mean preoperative IOP was 28.4 +/- 2.9 mm Hg in group 1 and 29.0 +/- 2.4 mm Hg in group 2. Final postoperative IOP was 17.0 +/- 1.6 for group 1 and 17.1 +/- 1.4 mm Hg for group 2. There were no significant differences between the groups for IOP, success rate, and complications (P > .05). The most common postoperative complication was shallow anterior chamber (11 [22.9%] of 48 eyes). CONCLUSIONS: A 0.4 or 0.2 mg/mL mitomycin C solution for 4 minutes may be used during trabeculectomy in refractory developmental glaucoma. PMID- 15825747 TI - The cost and yield of photoscreening: impact of photoscreening on overall pediatric ophthalmic costs. AB - BACKGROUND: Approximately 5% of preschool-age children suffer from amblyopia. Many of them have high or unequal hyperopia. Amblyogenic risk factors frequently can be detected by photoscreening. METHODS: Free photoscreening was offered to Alaskan children ages 1 to 5 from urban and rural screening hubs. Screened images were mailed to the Alaska Blind Child Discovery coordinating center for physician photoscreen interpretation, specifically seeking latent or anisometropic hyperopia. Parents and screeners then were mailed results and information about amblyopia. Follow-up examination data were tallied, and a cost-consequence analysis was developed for various vision screening paradigms and eye care. RESULTS: From 1996 through 2003, a total of 13,255 screenings were performed with a positive interpretation rate of 4.7%. Penetrance of screening was 22% in urban and 44% in rural communities. Positive predictive value was estimated to be more than 90%. Average cost to screen and inform an Alaskan preschooler was approximately 10.67 dollars, and cost to detect amblyogenic risk factors by photoscreening in an Alaskan was approximately 206 dollars. Compared to American Academy of Pediatrics (AAP) 1995 guidelines, implementing photoscreening added 9%, while mandating complete prekindergarten examination added 49% to overall eye care. CONCLUSIONS: MTI photoscreening achieved high community penetrance and high positive predictive value for latent hyperopia and other amblyogenic factors. When follow-up costs are considered, adding photoscreening to current AAP guidelines may add 112 dollars per child over 10 years, but probably would assist in the reduction of amblyopia. Penetrance of urban photoscreening likely will remain low unless pediatric vision screening guidelines and reimbursement are revised. PMID- 15825748 TI - Cone-rod dystrophy and acquired dissociated vertical nystagmus. AB - Although usually a manifestation of vestibular or neurological disease, acquired nystagmus occasionally can be seen in ocular diseases such as ocular albinism and cone dystrophy. The present case of cone-rod dystrophy presenting with acquired dissociated vertical nystagmus probably demonstrates another unusual manifestation of cone-rod dystrophy. PMID- 15825749 TI - Ocular complications of sigmoid sinus thrombosis from L-asparaginase. AB - This article presents a 9-year-old boy with acute lymphocytic leukemia who developed ocular complications after beginning treatment with L-asparaginase. Ocular motor examination revealed esotropia with a right abduction deficit, consistent with a sixth nerve palsy, and funduscopy revealed bilateral optic disc edema. Thrombosis of the right sigmoid sinus was noted on magnetic resonance imaging and magnetic resonance venography. Symptoms gradually resolved after discontinuing L-asparaginase and treatment with heparin. PMID- 15825750 TI - Optic nerve cysticercosis. AB - Optic nerve cysticercosis is rare but causes significant visual morbidity. A high index of suspicion is warranted for patients in endemic areas who present with sudden vision loss. Visual morbidity can be prevented by early imaging studies and therapeutic interventions. PMID- 15825751 TI - Congenital horner syndrome and hemiplegia secondary to carotid dissection. AB - A 5-month-old infant had right Horner syndrome and left hemiplegia. Magnetic resonance imaging confirmed smaller cerebral hemisphere and magnetic resonance angiography showed reduced blood flow in the internal carotid artery on the right. A diagnosis of congenital hemiplegia and carotid occlusion secondary to maternal trauma during pregnancy was made. PMID- 15825752 TI - Aniridia (PAX6(+/-)). PMID- 15825753 TI - New mosquito records of an area known for Japanese encephalitis hyperendemicity, Gorakhpur District, Uttar Pradesh, India. AB - During long-term ecological studies carried out between 1990 and 2000, a total of 67 mosquito species were collected from Gorakhpur District, Uttar Pradesh, India, of which 43 were documented for the 1st time. The genus Aedes was represented by 15 species, Anopheles by 15, Culex by 24, Mansonia by 3, and Mimomyia, Ochlerotatus, and Verrallina by 2 each. The genera Aedeomyia, Armigeres, Coquillettidia, and Uranotaenia were represented by single species. The impact of ecological changes on mosquito fauna in Gorakhpur District is discussed. The role of culicine mosquitoes in disease transmission in the area is highlighted. PMID- 15825754 TI - Crossing experiments of Anopheles minimus species C and putative species E. AB - In the Anopheles minimus complex, 2 sibling species (A and C) are generally accepted. Recently, a 3rd species, provisionally designated An. minimus species E, has been described from the Ryukyu Archipelago, Japan, based on crossing experiments (A and E), DNA analysis, mitotic karyotypes, and some morphological characteristics. The present study reports the results of crossing experiments between species C and putative species E. Hybridization between the progeny of An. minimus species C from Thailand and putative species E from Japan revealed postzygotic genetic incompatibility. Although F1 hybrid progeny were obtained from both directions of crosses, the hybrid males from C female x E male crosses were completely sterile, with atrophied testes and accessory glands. In addition, the external terminalia of all of these males never completely rotated and the males failed to copulate by artificial mating. In E female x C male crosses, the hybrid males showed partially sterile testes in which most spermatozoa were abnormal (enlarged head) and inactive, and they had very little success in inseminating females. The salivary gland polytene chromosomes of F1 hybrid larvae from species C female x species E male showed a fixed heterozygous inversion on the 3L arm. Those F1 hybrids from species E female x species C male showed partial asynapsis on identified arms (2R and 3L) and a fixed heterozygous inversion on the 3R arm. When the F1 hybrid females from both directions of crosses were backcrossed with either C or E males, they produced male progeny with abnormal spermatozoa. Study of mating behavior in a 30 x 30 x 30-cm cage showed that the C males failed to mate with either C or E females, indicating that species C males cannot breed in confined spaces (lack stenogamy). Putative species E males had little success in inseminating species C females. This study provides strong evidence of genetic incompatibility between An. minimus species C and putative species E, supporting previous data that species E is a distinct species in the An. minimus complex. PMID- 15825755 TI - New, recent, and questionable mosquito records from Mississippi. AB - Enhanced mosquito surveillance in Mississippi has confirmed the presence of Mansonia titillans and Ochlerotatus trivittatus, which represent new state records for Mississippi. Recent records of Aedes albopictus, Anopheles maverlius, An. quadrimaculatus, An. smaragdinus, and Ochlerotatus hendersoni, and questionable records of Ochlerotatus dorsalis, Oc. stimulans, and Psorophora pygmaea are reviewed. PMID- 15825756 TI - Investigation of relationships between Aedes aegypti egg, larvae, pupae, and adult density indices where their main breeding sites were located indoors. AB - Aedes aegypti (L.) density indices obtained in a dengue fever (DF) endemic area were compared. One hundred and twenty premises, in an urban area of Colombia where dengue type-1 and type-2 virus cocirculated, were randomly selected and sampled for 7 months. The geometric mean monthly numbers (density index, DI) of Ae. aegypti eggs (ODI), 4th instar larvae (LDI), pupae (PDI), and adults (ADI) were calculated based on the use of ovitraps, nets, and manual aspirators, respectively. A negative temporal correlation was observed between the LDI and the ODI (r = -0.83, df = 5, and P < 0.01). Positive temporal correlations were only observed between the LDI and the PDI (r = 0.90, df = 5, and P < 00.5) and the Breteau and House indices (r = 0.86, df = 5, and P < 0.01). No other correlations were found between these indices and any of the other density indices or the incidence of suspected DF cases in residents, the temperature, the rainfall, or seasonal fluctuations. Our results were, therefore, probably due to the most productive Ae. aegypti breeding sites (large water containers) being located indoors within this study area. The number of adult female Ae. aegypti/person (n = 0.5) and pupae/person (n = 11) in our study area were lower and dramatically higher than the transmission thresholds previously reported for adult and pupae, respectively. Because there were confirmed DF cases during the study period, the transmission threshold based on the Ae. aegypti pupae was clearly more reliable. We found that the mean ovitrap premise index (OPI) was 98.2% during this study and that the mean larval (L-4th instars) premise index (LPI) was 59.2%, and therefore we suggest that the OPI and LPI would be more sensitive methods to gauge the effectiveness of A. aegypti control programs. PMID- 15825757 TI - Vegetation management to stimulate denitrification increases mosquito abundance in multipurpose constructed treatment wetlands. AB - Inundation of dried emergent vegetation that has been knocked down using heavy equipment is a vegetation management strategy that provides an inexpensive source of supplemental organic carbon to stimulate denitrification, a process important for nitrate removal in constructed treatment wetlands. The abundance of larval mosquitoes (Culex sp. and Culiseta sp.) in multipurpose constructed wetlands that had undergone autumnal vegetation management intended primarily to enhance denitrifying bacteria was significantly higher than in wetlands that remained in continuous operation. Mosquito abundance in wetlands that had undergone vegetation management during autumn was much lower than that observed during spring in a previous study; nevertheless, vegetation management practices that inundate dried, felled macrophytes are contraindicated for mosquito control. PMID- 15825758 TI - Colonization of Ochlerotatus taeniorhynchus from Riohacha, Colombia. AB - The main objective of the present work was to establish and maintain a colony of Ochlerotatus taeniorhynchus, Riohacha strain, under laboratory conditions and to study its life cycle. The colony's insectary was established from adult male and female mosquitoes collected from peripheral neighborhoods of Riohacha, Colombia. Environmental conditions established in the insectary were 26.5 degrees C average temperature, 80% average relative humidity, and 12 h photoperiodicity. Eight continuous generations were taken into account for maintaining the mosquitoes and analyzing their life cycle. The male mosquito's average life cycle was 26.8 days. The female's cycle was 30.8 days. Analysis of each of the biological stages of development (mean days) produced the following results: egg incubation 4.55 +/- 0.291, larvae 8.28 +/- 0.499, pupae 1.32 +/- 0.215, adult male 12.65 +/- 5.920. and adult female 16.73 +/- 6.034. The Riohacha colony has been maintained for 32 generations in 31 months. Comparison of the Riohacha colony with the previously established Cartagena and Barranquilla colonies showed few differences in the duration of stages of the life cycle between strains. PMID- 15825759 TI - Impact of West Nile virus outbreak upon St. Tammany Parish Mosquito Abatement District. AB - St. Tammany Parish, Louisiana, experienced an outbreak of West Nile virus (WNV) in 2002, with 40 human cases and 4 deaths, most occurring from June to August. Culex pipiens quinquefasciatus was believed to be the primary vector of WNV during the outbreak, although circumstantial evidence suggests that Aedes albopictus also may have been involved in transmission. Dead bird reports were the 1st indication of the outbreak and were an excellent indicator of WNV activity; however, sentinel chickens were the most effective in tracking viral activity. Although sentinel chickens tested positive 2-3 wk after the 1st dead bird, they peaked at about the same time as human cases, and continued testing positive when viral activity was no longer detected in birds and mosquito pools. Lag time between the 1st positive sentinel chicken and the peak in human cases was 6 wk. If this trend continues in the future, sentinel chickens could be used to predict the peak in number of human cases. The 2002 WNV outbreak had a strong impact on operational budget of the St. Tammany Parish Mosquito Abatement District (88% increase above the 3-year average). Vector control activities accounted for most of the operational increase and consisted of targeted population reduction of known WNV-competent mosquito species. The goal of these activities was to prevent new human WNV cases. The 3- to 10-fold reduction in vector mosquito populations from May to August, together with a dramatic drop in number of new human cases by the end of August, indicated that our strategy was effective. PMID- 15825760 TI - Detection of chicken antibodies to mosquito salivary gland antigens by enzyme immunoassay. AB - Sentinel chickens are used to detect western equine encephalomyelitis, St. Louis encephalitis, and West Nile virus activity. Flocks that receive high mosquito exposure will be most effective for surveillance purposes. However, mosquito population indices at the flock sites may only provide an indirect measure of potential exposure. Therefore, we developed an indirect enzyme immunoassay to detect chicken antibodies to salivary gland antigens (SGAs) from Culex tarsalis, the primary arbovirus vector in California. Chickens fed upon by Cx. tarsalis developed an antibody response that was proportional to the amount of exposure. Cross-reactivity between sera from Cx. tarsalis-exposed chickens and SGAs from Culex pipiens quinquefasciatus, Culex pipiens pipiens, Ochlerotatus melanimon, and Ochlerotatus sierrensis was likely due to shared SGAs among these species. This serologic assay for mosquito exposure could be used to evaluate the sensitivity of sentinel flocks for detecting arboviral activity. PMID- 15825761 TI - Quantification of West Nile virus in vector mosquito saliva. AB - Saliva was collected from 4 species of mosquitoes intrathoracically inoculated with West Nile virus (WNV). The amount of infectious virus in the saliva was quantified by plaque assay and the number of WNV genomic equivalents (GE) was measured by reverse transcriptase-polymerase chain reaction. Ochlerotatus triseriatus had the greatest mean amount of infectious virus per saliva collection, followed by Aedes albopictus, Culex pipiens, and Cx. quinquefasciatus. The mean GE/saliva collection was also greatest in Oc. triseriatus, followed by Cx. quinquefasciatus, Cx. pipiens, and Ae. albopictus. The variance of log GE/saliva collection for Ae. albopictus was significantly lower than the variance for the other 3 species. This study provides a basis for comparing this component of vector competence and for determining the amounts of virus inoculated into vertebrates in experimental host competence studies. PMID- 15825762 TI - Persistence and wash-resistance of insecticidal efficacy of nettings treated with deltamethrin tablet formulation (K-O TAB) against malaria vectors. AB - Persistence, wash-resistance, and shelf life of mosquito nets treated with a water-dispersible tablet formulation of synthetic pyrethroid insecticide deltamethrin (K-O TAB) at 25 mg/m2 was evaluated against malaria vectors in India. During June 2001, treated and untreated polyester, nylon, and cotton nets were separately distributed in 3 villages and cone bioassays were performed on Anopheles culicifacies and An. stephensi 1 day after treatment and thereafter every month for 12 months. The mosquitoes were exposed for 3 min on the nettings (treated and unwashed, or treated and washed once or twice in 3 months, and untreated) and knock-down (1 h) and 24 h postexposure mortality were recorded. Unwashed polyester nets, and those washed once 1 month after treatment, gave 100% mortality in An. culicifacies for 6 months. A 2nd wash at 3 months after treatment marginally reduced the insecticidal action. Anopheles stephensi was fully susceptible up to 4 months when exposed to unwashed nets but washing considerably reduced insecticidal action (65-78% after 2 washes). Treated nylon and cotton nets were effective for 4 months on both vectors. Treated nets kept on shelf retained 100% efficacy for 10 months. Overall, the treated nets gave a considerably long persistence of insecticidal action even after a single wash. Treated polyester nets were found most effective. Compared with our earlier experiences of using liquid formulations, the tablet formulation is likely to have a better community acceptance in treating nets. PMID- 15825763 TI - Impact of environmental factors on the toxicity of Bacillus thuringiensis var. israelensis IPS82 to Chironomus kiiensis. AB - To determine the feasibility of microbiological control of chironomid larvae in water sources for the city of Shenzhen, China, the toxicity characteristics of Bacillus thuringiensis var. israelensis (Bti) IPS82 on Chironomus kiiensis Tokunaga were studied. Tests on fermentation products of IPS82 showed good correlations between toxicity, cell density, dissolved oxygen, and the spore forming phase. In this study, bioassays were carried out with different stage larvae. Results showed that the LC50s (24 h postexposure) were 8.2, 15.2, 24.7, and 38.6 mg/liter for the 1st, 2nd, 3rd, and 4th instars, respectively. Tests on environmental factors influencing toxicity of Bti to C. kiiensis showed that sunlight is the most important factor, shortening the half-life of Bti from 21 days in dark to 10 days under sunlight. Temperature variations (15-30 degrees C) caused no impact on toxicity, but a 16% increase in larval mortality was observed at 35 degrees C. The toxicity of IPS82 was greatest at a pH of 7. In field trials, dosages above 100 mg/liter were effective in the control of C. kiiensis. Our study indicated that it is feasible to use Bti to control C. kiiensis in city source water. PMID- 15825764 TI - A new in vitro bioassay system for discovery of novel human-use mosquito repellents. AB - A Klun & Debboun (K&D) test module, previously developed and used for quantitative measurement of the efficacy of mosquito repellents on human volunteers, was adapted for in vitro evaluation of repellents by coupling the module with a membrane-blood reservoir. Performance of Deet, Bayrepel, and SS-220 insect repellents in the new in vitro system was compared with their performance on humans against mosquitoes using our standard in vivo system. For each compound, in vitro dose-response assays were conducted with compounds applied to cloth positioned over blood reservoirs covered with Baudruche membrane against Aedes aegypti. The repellents were also tested in vitro against Anopheles stephensi and Ae. aegypti at a fixed dose of 24 nmol compound/cm2 cloth over the Baudruche and Edicol collagen membranes. Concurrently, the repellents were tested at the fixed dose using the K&D module on human volunteers. The observed proportions of both mosquito species deterred from biting in the fixed doses in the in vitro assays were similar to those obtained using humans, being clearly able to distinguish controls from repellents, and differing only in the ranking of the effectiveness of some of the repellents. Dose-response relationships of the in vitro and in vivo systems were also very similar, although not directly comparable because the data were not collected concurrently. This new in vitro assay system can be used in high throughput screening of compounds to identify new repellents having potential for use as topical mosquito repellents on humans. PMID- 15825765 TI - Cytochrome P450 genes: molecular cloning and overexpression in a pyrethroid resistant strain of Anopheles minimus mosquito. AB - We previously determined that physiological resistance in a laboratory-selected pyrethroid-resistant Anopheles minimus species A Theobald mosquito is associated with increased detoxification via a P450-mediated mechanism. A CYP6 gene, CYP6AA3, was subsequently cloned and found overexpressed in 2 resistant mosquito generations (F13 and F19). We report herein the cloning of CYP6P7 and CYP6P8 genes with full coding sequences from the same An. minimus mosquito colony strain. CYP6P7 and CYP6P8 encode proteins, each with 509 amino acids. CYP6P7 had the closest (81%) amino acid identity with Anopheles gambiae CYP6P2. CYP6P8 genes had 79% identity with An. gambiae CYP6P1. Using semiquantitative reverse transcription-polymerase chain reaction analysis, the mRNA expression level of CYP6P7 presented approximately 2- and 4-fold increases in F19 and F25 deltamethrin-resistant populations, respectively, compared with the parent susceptible strain. CYP6P8 mRNA expression levels were not significantly different between the 3 filial generations. The overexpression of CYP6AA3 mRNA was greater than that of CYP6P7 in F19 and F25 resistant populations. The relative increase of both CYP6AA3 and CYP6P7 mRNA was correlated with increased resistance to deltamethrin in An. minimus. PMID- 15825766 TI - Monoterpenes from thyme (Thymus vulgaris) as potential mosquito repellents. AB - Five monoterpenes (carvacrol, p-cymene, linalool, alpha-terpinene, and thymol) derived from the essential oil of thyme (Thymus vulgaris) were examined for their repellency against the mosquito Culex pipiens pallens. All 5 monoterpenes effectively repelled mosquitoes based on a human forearm bioassay. Alpha terpinene and carvacrol showed significantly greater repellency than a commercial formulation, N,N-diethyl-m-methylbenzamide (deet), whereas thymol showed similar repellency to that of deet. The duration of repellency after application for all these monoterpenes was equal to or higher than that of deet. These findings indicate that a spray-type solution containing 2% alpha-terpinene may serve as an alternative mosquito repellent. PMID- 15825767 TI - Integration of Bacillus thuringiensis H-14 formulations and pyriproxyfen for the control of larvae of Aedes aegypti and Aedes albopictus. AB - Studies were carried out on the bioefficacy and residual activity of Bacillus thuringiensis israelensis H-14 (Bti) (water-dispersible granules of VectoBac ABG 6511 and liquid formulations of VectoBac 12AS) and pyriproxyfen (insect growth regulator, Sumilarv 0.5%) as direct applications for control of larvae of Aedes aegypti and Aedes albopictus. Two dosages of each Bti formulation (285 and 570 international toxic units [ITU]/liter) and the integration of both Bti formulations and pyriproxyfen were used for residual tests with 45-liter earthen jars for a period of 4 wk. In 1 test series, the treated water was replenished daily with 6 liters of seasoned untreated water. In the 2nd test series, the water in the jars was topped up to the 40-liter level during evaluation. Neither Bti formulation remained effective for a full week. Water-dispersible Bti granules provided effective initial control activity against Ae. aegypti and Ae. albopictus for both test designs (with replenishment and without replenishment of water). The higher dosage (570 ITU/liter) for both Bti formulations was only partially effective at the end of 1 wk after being diluted. After 1 wk, water dispersible Bti granules provided greater larval mortality than did liquid Bti formulation against both mosquito species when integrated with pyriproxyfen. Pyriproxyfen (79.5 and 159 mg/liter) on its own showed low larvicidal activity but provided very effective control of adult emergence. In this study, integration of Bti (285 and 570 ITU/liter) with pyriproxyfen (79.5 mg/liter) extended the duration of partial larval control somewhat, but live larvae persisted throughout the 4-wk test. The integration effect was more obvious when water-dispersible Bti granules were integrated with pyriproxyfen than when liquid Bti was used. Integration of Bti with pyriproxyfen had a negative effect on adult emergence, which was completely inhibited by pyriproxyfen after day 1. Daily replenishment of water increased Bti activity and provided slightly better larval control. Aedes albopictus and Ae. aegypti were both completely susceptible to the higher concentration of Bti and pyriproxyfen in both test designs (with replenishment and without replenishment of water). PMID- 15825769 TI - The larval habitat of Verrallina (Verrallina) lugubris and chaetotaxy of field collected larvae. AB - The larval habitat of Verrallina lugubris (Barraud) is defined based upon collection of 648 larvae from swamp pools from January to March 1998 in Pichavaram mangrove forest, and 125 larvae during September 1997 in Maravakadu mangrove forest in the eastern coast of South India. Salinity of the larval habitat ranged from 12,150 to 24,500 mg/liter in Pichavaram and from 11,600 to 13,000 mg/liter in Maravakadu. Associated mosquito species were Culex sitiens Wiedemann, Ochlerotatus portonovoensis (Tewari and Hiriyan), and Anopheles subpictus Grassi. The chaetotaxy of field-collected larvae of Ve. lugubris is presented. PMID- 15825768 TI - Culicoides and other biting flies on the Palos Verdes Peninsula of Southern California, and their possible relationship to equine dermatitis. AB - Biting insects were sampled to investigate the cause(s) of dermatitis (putative Culicoides hypersensitivity [CHS]) on horses on the Palos Verdes Peninsula, Los Angeles County, California. Suction traps baited with CO2 were operated at 5 sites from April 2002 to August 2003, supplemented by sampling from horses. Six species of Culicoides, 7 species of mosquitoes, and occasional Simulium and Stomoxys were collected in traps. Culicoides sonorensis was the most common midge trapped, although C. obsoletus and C. freeborni also were collected repeatedly. Insects from the belly region of horses in spring (peak dermatitis period) were mostly C. obsoletus, which were far more abundant than indicated by CO2-baited suction traps. The predominant mosquito was Culiseta incidens; Culex quinquefasciatus and Cx. tarsalis also could be common, depending on the site. Mosquitoes, particularly Cs. incidens, were more abundant the 2nd year (normal rainfall), whereas C. sonorensis and C. freeborni were more abundant the 1st year (drought period). Culicoides obsoletus, supplemented by C. sonorensis, is regarded as a primary suspect causing CHS in this area. PMID- 15825770 TI - Update on Ochlerotatus japonicus in the state of Washington. AB - A limited survey was conducted in 2003 to determine if Ochlerotatus japonicus was spreading outside of King County, Washington. Ochlerotatus japonicus was found in north-central Pierce County and west-central Snohomish County. These collections represent county records. An additional site was reported from King County. Ochlerotatus japonicus is established and spreading to other areas of Washington. PMID- 15825771 TI - The first record of larval collections of Ochlerotatus thibaulti in New York state. AB - Larvae of Ochlerotatus thibaulti Dyar and Knab have not been reported in New York State or New England, but adult records suggest that breeding populations are present. In 2003 and 2004, larval Oc. thibaulti were collected at 3 sites upstate near the Catskill Mountain region in Lomontville, NY. This is the 1st record of Oc. thibaulti collected in New York and New England. Larvae were found in a variety of subterranean habitats within and near semipermanent woodland pools, including under hummocks and in deep and shallow depressions of upturned trees. No immatures were observed after late June. Larvae of Culiseta melanura were found in the same habitats in the early spring and later in the season after Oc. thibaulti had emerged. PMID- 15825772 TI - First report of West Nile virus in mosquitoes from Lubbock County, Texas. AB - Since July 2002, ongoing surveillance efforts have been conducted to determine potential vectors of West Nile virus (WNV) and Saint Louis encephalitis virus (SLEV) in the mosquito population occurring in Lubbock County, Texas. Adult mosquitoes collected in Lubbock County during 2002 and 2003 represented 7 genera, with Culex tarsalis and Ochlerotatus sollicitans being the predominant species collected. Mosquitoes were initially screened for WNV and SLEV by using the VecTest antigen panel assay. Positive VecTest results were confirmed by reverse transcriptase-polymerase chain reaction. West Nile virus-positive pools of mosquitoes were detected in 2002 and 2003, with the majority of the positive pools consisting of Cx. tarsalis. None of the mosquito pools tested positive for SLEV. PMID- 15825773 TI - Aerial optimization and canopy penetration study of Dibrom 14 Concentrate. AB - This document describes the results of collaborative trials between St. Tammany Parish Mosquito Control, ADAPCO, Inc., and the Navy Disease Vector Ecology and Control Center, Naval Air Station, Jacksonville, FL. These tests provide data on the aerial optimization testing of Dibrom 14 Concentrate adulticide. During the week of June 17, 2002, 1 canopy penetration and 3 optimization studies were conducted in St. Tammany Parish, LA, using a Britton Norman twin turbine Islander equipped with 10-800067 flat fan nozzles. Dibrom 14 Concentrate (AMVAC Chemical Corp., Los Angeles, CA) was applied at 3.12 liter per min from a release height of 60.96 m above ground level (agl) at 140 kts indicated airspeed. This resulted in an application rate of 0.65 oz/acre. PMID- 15825774 TI - Design of a mosquito trap support pole for use with CDC miniature light traps. AB - A mosquito trap support pole constructed from polyvinyl chloride and aluminum pipes was designed to hang a Centers for Disease Control and Prevention miniature light trap and dry ice container. Miniature light traps normally hang from tree branches. The trap support pole is designed to hang traps and dry ice bait in areas where no suitable trees exist. PMID- 15825775 TI - Advanced orthodontic education: evolution of assessment criteria and methods to meet future challenges. AB - The achievement of academic and research institutes has increasingly become the subject of ranking by independent organizations. This trend has educational and economic implications, which may pertain to attracting students and endowments as well as allocating of funds to specific educational components. The aims of this article were to discuss the reliability and soundness of the existing academic ranking methods and to propose guidelines for the quantitative assessment of advanced orthodontic programs. This criteria-driven assessment assigns weight factors for the basic components of orthodontic education and provides a template for clarifying the standing of programs. Moreover, a combination of program and individual assessment is proposed to monitor the implementation of educational criteria and assess their effectiveness. PMID- 15825776 TI - Long-term dental arch changes after rapid maxillary expansion treatment: a systematic review. AB - This systematic review evaluates long-term dental arch changes after rapid maxillary expansion treatment on orthodontic patients with constricted arches. Clinical trials that assessed dental arch changes through measurements on dental casts or cephalometric radiographs were selected. No patients with surgical or other simultaneous treatment during the active expansion period were accepted. Electronic databases were searched with the help of a senior Health Sciences librarian. Original articles were retrieved from the selected abstracts, and their references were also scanned for possible missing articles. Forty-one articles met the initial inclusion criteria, but 35 were later rejected because they lacked a control group or only evaluated dental changes or used a semirapid technique. Some of them also lacked a reported measurement error. From the remaining articles, two did not report a long-term evaluation. From the final four articles, two measured changes through dental casts and two assessed changes through radiographs (one through lateral cephalometric radiographs and one through posteroanterior radiographs). Similar maxillary molar and cuspid expansion could be found in adolescents and young adults. Significantly less indirect mandibular molar and cuspid expansion was attained in young adults compared with adolescents. A significant overall gain in the maxillary and mandibular arch perimeter was found in adolescents. More transverse dental arch changes were found after puberty as compared with before, but the difference may not be clinically significant. No anteroposterior dental changes were found on lateral cephalometric radiographs. PMID- 15825777 TI - Assessment of orthodontic treatment outcomes: early treatment versus late treatment. AB - This investigation compares the treatment outcome of early treatment (in the mixed dentition) with that of late treatment (early permanent dentition) using objective evaluation criteria. Pretreatment and post-treatment records of all patients (n = 512) completed from 1998 to 2000 in the graduate orthodontics clinic at the Indiana University School of Dentistry (IUSD) were evaluated by the American Board of Orthodontics Objective Grading System (ABO OGS) and Comprehensive Clinical Assessment (CCA) method developed at IUSD. Two definitions of early treatment were used in this study: (1) all patients started in the mixed dentition with early-treatment objectives and (2) female individuals were <10 years and male individuals were <10.5 years of age when treatment began. Comparison of the final results between early- vs late-treatment groups showed that the early-treatment group had significantly longer treatment time and worse CCA scores than the late-treatment group, regardless of the definition of the early-treatment group or whether the early-debond (premature treatment termination) cases were included or not. There was no significant difference between early- and late-treatment groups regarding the ABO OGS score, which indicated that the CCA method is more sensitive in detecting compromised outcomes for patients with long treatment times. Prematurely terminated treatment was more prevalent in the early-treatment group than in the late-treatment group. In this large sample of consecutive patients (n = 512), the disadvantages of early treatment was prolonged treatment time, worse CCA score, and a higher incidence of premature termination of treatment, which was attributed to patient/parent "burn-out." PMID- 15825778 TI - Changes in head posture after rapid maxillary expansion in mouth-breathing girls: a controlled study. AB - The influence of respiratory function on craniofacial development and head posture has been demonstrated previously. This study evaluated the effect of rapid maxillary expansion (RME) on nasopharyngeal airway adequacy, head posture, and facial morphology in children with nasal obstruction. Fifty-five girls (8-15 years of age) who needed maxillary expansion, showed reduced nasopharyngeal airway adequacy (pm-Ad 2), and were subjectively assessed as mouth breathers were allocated randomly into 2 groups. The 23 subjects in the first group were treated with RME, and the 22 subjects in the other group were followed about 8 months before beginning therapy and became untreated controls. Dental casts and lateral skull radiographs exposed in natural head position were obtained at the first visit and 6 months later for all subjects. In the girls under active treatment there was a statistically significant increase of pm-Ad 2 (P < .0001), a significant increase of the cervical lordosis angle (P < .0001), a flexion of the head (P < .0001), and a decrease in the craniocervical angulation (P < .0001) (paired t-tests). No significant changes were seen in the control group. The correlation coefficients indicated a mild correlation between pm-Ad 2 distance and craniocervical angulation (SN/OPT angle) (r = 0.61 at P < .001). RME is capable of increasing nasopharyngeal airway adequacy in girls, and this leads to a decrease in craniocervical angulations. The clinical importance of these results is yet to be clarified. PMID- 15825779 TI - First or second premolar extraction effects on facial vertical dimension. AB - If the concept of mesial movement of molars to reduce the "wedge effect" and decrease facial vertical dimension (FVD) is valid, it is important to investigate the effect of first (P1) and second premolar (P2) extraction on FVD. This study compares the mesial movement of molars and changes in FVD between P1 and P2 extraction groups in Class I malocclusion with a hyperdivergent facial type. We compared 27 cases with maxillary and mandibular P1 extractions (group 1) and 27 cases with maxillary and mandibular P2 extractions (group 2). To determine FVD changes due to treatment and to compare differences between two groups, paired t test and independent t-test were performed, respectively. Group 2 showed more mesial movement of the maxillary and mandibular first molars and less retraction of the maxillary and mandibular central incisors than group 1 (P < .05). Both groups showed increased anterior facial height (P < .05), but there were no statistically significant differences in angular and proportional measurements between pre- and posttreatment. There was no significant difference in the amount of FVD change between groups 1 and 2 except in the maxillomandibular plane angle and SN to palatal plane angle (P < .05). These results suggest that there is no decrease in FVD regardless of the maxillary and mandibular P1 or P2 extraction. Therefore, the hypothesis that P2 extraction in hyperdivergent facial types will result in mesial molar movement and decrease FVD by reducing the wedge effect is invalid. PMID- 15825780 TI - Association of malocclusion and functional occlusion with subjective symptoms of TMD in adults: results of the Study of Health in Pomerania (SHIP). AB - An analysis of exclusively representative population-based studies on adults has shown that only few and inconsistent associations could be detected between malocclusions and clinical signs of temporomandibular disorders (TMD)--and none for functional occlusion factors (occlusal interferences, non-working side contacts, etc). The aim of this study was to analyze associations between morphologic occlusion as well as factors of functional occlusion and subjectively perceived symptoms of TMD--again on the basis of the population-based Study of Health in Pomerania (SHIP), providing a sample of 4310 subjects (out of 7008 subjects yielding a response rate of 68.8%) aged 20 to 81 years, and other international representative studies from the systematic review. Besides occlusal factors also parafunctions and socioeconomic status (SES) were taken into account (including age and sex). Multiple logistic regression analysis was used--adjusted for SES. In this study, none of the occlusal factors were significantly associated with the indication of more frequent subjective TMD symptoms. However, the parafunction "frequent clenching" was connected with subjective TMD symptoms (odds ratio = 3.4). Compared with other population-based studies few and (across studies) inconsistent associations between malocclusions and subjective TMD symptoms could be ascertained. No significant associations of factors of functional occlusion with TMD symptoms were identifiable. PMID- 15825781 TI - A special method of predicting mandibular growth potential for Class III malocclusion. AB - The purpose of this study was to establish an equation to predict the mandible growth potential (GP) for Class III malocclusion on the basis of the analysis of the cervical vertebrae in a single cephalometric radiograph and to compare its predictive accuracy with other methods. Data comprised two groups each with 22 Japanese girls. Group A was examined to construct the prediction equation. Group B served to compare the predictive accuracy with the GP method and the method of Mito et al (MM). The following results were obtained: (1) an equation was determined to obtain mandible GP on the basis of measurements in the third and fourth cervical vertebral bodies and (2) the average errors between the predicted increment and the actual increment for each method were 1.45 mm for the equation, 2.91 mm for the GP method, and 2.48 mm for the MM. These results suggest that using cervical vertebral measurements might allow predicting the mandible GP length for Class III malocclusion. PMID- 15825782 TI - Ossification of laryngeal cartilages on lateral cephalometric radiographs. AB - This study was undertaken to identify the variations in the physiologic ossification of human laryngeal cartilages by evaluating the lateral cephalometric radiographs of healthy males and females referred to our facility. Lateral cephalometric radiographs of 359 patients (141 male and 218 female; ages 10 to 59 years) referred from various specialty clinics between April 1998 and March 2000 were used. Frequencies and confidence intervals were obtained and tabulated for both thyroid and cricoid ossifications as seen on these cephalometric radiographs. The interobserver agreement was strong for both thyroid (kappa = .986) and cricoid (kappa = .982) observations. Overall, thyroid cartilage was more frequently ossified compared with cricoid. Various degrees of thyroid and cricoid cartilage ossification were found in 186 patients (120 female and 66 male) in the third decade and beyond. Among 173 patients in the first 2 decades (98 females and 75 male), evidence of thyroid ossification was found in 2 male patients aged 14 and 18, and ossification was detected in the cricoid cartilage in 12 patients. There was a preponderance of laryngeal cartilage ossification in men compared with women. Radiographically detectable laryngeal ossification increased with age starting in the third decade. There is a general trend of increase in the ossification of laryngeal cartilages as the age advanced. PMID- 15825783 TI - Are the lower incisors the best predictors for the unerupted canine and premolars sums? an analysis of a Peruvian sample. AB - The lower permanent incisor tooth width sum has been proposed as the best predictor for the tooth width sums of the unerupted canine and premolars (SPCP) for populations from different ethnic origins. Only two previous studies have refuted it. The purpose of the present study was to determine which sum or combination of sums of permanent tooth widths presented the best prediction capability for the SPCP in a Peruvian sample, to calculate a specific linear regression equation for this population, and to evaluate the clinical significance. A total of 150 children with complete permanent dentitions were selected. Fifty more children were used as a validation sample for the application of a multiple linear regression equation (MLRE). They did not present clinically visible dental caries or proximal restorations and no active or previous orthodontic treatment. Their dental casts were measured to 0.1 mm with a sliding caliper with a Vernier scale. Three-way analysis of variance, Pearson Correlation Test, Fisher Z values and a MLRE were used for the statistical analysis. The combination of the sums of permanent upper and lower central incisors and upper first molars was the best predictor for the SPCP in this sample. A MLRE was calculated including sex and arch as additional predictor variables. The MLRE determination coefficient was 60% with a standard error of 0.8 mm. This new MLRE underestimates (less than 1 mm discrepancy) the actual SPCP in only 7% of the cases on the basis of a validation sample. PMID- 15825784 TI - Intermaxillary tooth size discrepancy and malocclusion: is there a relation? AB - The aims of this study were to identify the possible sex differences in tooth size ratios between males and females, to determine whether there is a difference in the incidence of tooth size discrepancies for both the anterior and overall ratios when comparing with Angle Class I; Class II, division 1; Class II, division 2; and Class III malocclusion groups, to compare the tooth size ratios of different malocclusion groups with the anterior and overall tooth size ratios of 150 untreated normal occlusion subjects. In addition, the aim was to determine the percentage of tooth size discrepancies outside 2 SD from Bolton means for tooth ratios present in each malocclusion group and in the overall sample of this study. This study consisted of 150 subjects who served as the normal occlusion group and 560 patients who showed four different malocclusion characteristics (Angle Class I; Class II, division 1; Class II, division 2; and Class III). Tooth size measurements were performed on the models of normal occlusion and pretreatment models. For statistical evaluation, Student's t-test, analysis of variance and Tukey Honestly Significant Difference tests were performed. A significant sex difference was found only in the overall ratio for normal occlusion subjects (P < .001). All malocclusion groups showed statistically significant higher overall ratios than the normal occlusion group (P < .001). There were no statistically significant differences among malocclusion groups; however, there were a large number of patients within each group who had discrepancies greater than 2 SD from the mean. Further investigations are needed to explain the probable racial differences and relationships between malocclusion and tooth size measurements. PMID- 15825785 TI - Pain control during fixed orthodontic appliance therapy. AB - The control of pain during orthodontic treatment is of great interest to both clinicians and patients. However, there has been limited research into the control of this pain, and there is no standard of care for controlling this discomfort. This prospective study determines the pain sequelae in fixed orthodontic treatment and evaluates comparatively the analgesic effects of nonsteroidal anti-inflammatory drugs for the control of this pain. One hundred and fifty orthodontic patients who were to have teeth bonded in at least one arch were randomly assigned to one of six groups: (1) placebo/placebo, (2) ibuprofen/ibuprofen, (3) flurbiprofen/flurbiprofen, (4) acetaminophen/acetaminophen, (5) naproxen sodium/naproxen sodium, and (6) aspirin/aspirin. The pain evaluations were made during chewing, biting, fitting the front teeth, and fitting the back teeth using a 100-mm visual analogue scale (VAS) for seven days. All the analgesics succeeded in decreasing the pain levels compared with the placebo group. However, naproxen sodium and aspirin groups showed the lowest pain values, and the acetaminophen group showed VAS results similar to those of the two analgesics. PMID- 15825786 TI - Thermal changes in the pulp chamber during different adhesive clean-up procedures. AB - The aim of this in vitro study was to measure the temperature changes in the pulpal chamber when different adhesive clean-up procedures were used. Ninety intact extracted human maxillary central incisors were used in the study. The teeth were divided into six groups of 15 teeth each. The removal of the remaining composite on the tooth surface was performed with a tungsten carbide bur. The residual adhesive was removed using a high-speed handpiece with and without water cooling and a contra-angle handpiece with and without water cooling at high and low speeds. A J-type thermocouple wire was positioned in the center of the pulp chamber. The results were analyzed with analysis of variance (ANOVA) and the Tukey-honestly significant difference test. Two-factor ANOVA revealed significant interaction between the handpiece type and water cooling. In this study, the high speed contra-angle handpiece without water cooling group had the highest deltaT values (7.58 degrees C+/-1.84 degrees C) among all the clean-up procedures. The decrease in pulpal temperature with water cooling was -5.34 degrees C for the handpiece, -5.36 degrees C for the low-speed contra-angle handpiece and -4.98 degrees C for the high-speed contra-angle handpiece. Clinicians should be aware of the potential thermal damage to the pulp, which may result from long clean-up procedures without water cooling. Adhesive removal procedures should be performed with adequate water cooling to prevent temperature increases that might be harmful to pulpal tissues. PMID- 15825787 TI - Measurement of white lesions surrounding orthodontic brackets: captured slides vs digital camera images. AB - Digital images provide an efficient means of processing and storing clinical images and are increasingly being used in orthodontic practice. However, for research purposes it must be shown that measurements from digital images are as reliable as those from captured slides, particularly in the area of postorthodontic demineralization. The aim of this in vitro study was to compare the assessment of demineralized white lesions surrounding orthodontic brackets using images produced from captured slides with those from a digital camera. Thirty teeth with orthodontic brackets and a systematic arrangement of artificially induced demineralization were used. Standardized images of the teeth were taken in 35-mm slide format and using a digital camera. The slides were scanned and saved as digital images. All the images were numbered, recoded in a random order, and assessed by a second investigator. The image was examined for the presence or absence of demineralization. This was compared with the actual demineralization pattern, and the positive and negative predictive values were calculated. Twenty images from each group were randomly chosen and duplicated for an assessment of measurement error. The positive and negative predictive values were better from the digital images (0.92 and 0.81) than from the captured slides (0.88 and 0.74). The percentage agreements for the repeat assessments of the same slide were similar (96% for the captured slides and 93% for the digital images). Measurements of enamel demineralization using images from a digital camera are as accurate and reproducible as images captured from a photographic slide. PMID- 15825788 TI - Archwire ligation techniques, microbial colonization, and periodontal status in orthodontically treated patients. AB - Fixed or removable orthodontic appliances impede the maintenance of oral hygiene and result in plaque accumulation. Plaque retention surrounding orthodontic appliances leads to enamel demineralization caused by organic acids produced by bacteria in the dental plaque. Many studies have evaluated the effects of fixed orthodontic appliances on microbial flora and periodontal status, but only a few have evaluated the method of ligation as an additional factor. The aim of this study was to determine the changes in microbial flora and periodontal status after orthodontic bonding and to determine whether two different archwire ligation techniques affect these changes. A total of 21 orthodontic patients scheduled for fixed orthodontic treatment were selected for this split-mouth study. Two commonly used auxiliaries (elastomeric rings and ligature wires) for tying archwires were tested. Microbial and periodontal records were obtained before bonding (T0), one week later (T1), and five weeks after bonding (T2). Paired t-test and Wilcoxon signed rank test were used to compare the groups statistically. Although, teeth ligated with elastomeric rings exhibited slightly greater numbers of microorganisms than teeth ligated with steel ligature wires, the differences were not statistically significant and could be ignored. The two archwire ligation techniques showed no significant differences in the gingival index, bonded bracket plaque index, or pocket depths of the bonded teeth. However, teeth ligated with elastomeric rings were more prone to bleeding. Therefore, elastomeric ring use is not recommended in patients with poor oral hygiene. PMID- 15825789 TI - Comparison of bonding time and shear bond strength between a conventional and a new integrated bonding system. AB - Conventional adhesive systems use 3 different agents, an enamel conditioner, a primer solution, and an adhesive resin during the bonding of orthodontic brackets to enamel. A characteristic of some new bonding systems is that they combine the conditioning and priming agents into a single application as well as precoat the bracket with the adhesive in an attempt to save time during the bonding procedure. This study compared the total bonding time and shear bond strength (SBS) of 2 bracket-bonding systems: (1) an integrated system that incorporates a self-etching primer and precoated brackets and (2) a conventional system in which the etchant and primer are applied separately and the adhesive applied to the bracket by the clinician. The results of the SBS and the total bonding time comparisons (t = 3.451) of the 2 adhesive systems showed a significant difference (P = .0001). The mean SBS was 9.4+/-3.7 MPa for the new bonding system and 6.2+/ 4.4 MPa for the conventional system. The mean total bonding time was 36.5 s/tooth for the new system and 46.7 s/tooth for the conventional system. The clinician has to decide whether the increase in bond strength, the decrease in the total bonding time, and the steps saved during the bonding procedure with the new bonding system balance the increased cost incurred. PMID- 15825790 TI - Effects of using a new protective sealant on the bond strength of orthodontic brackets. AB - This study determines the effect of applying a new protective sealant to the enamel surface on the shear bond strengths of orthodontic brackets. Sixty teeth were randomly divided into three groups. In group 1 (control), the brackets were bonded to the etched teeth using the regular sealant. In group 2, the sealant was replaced with Pro Seal and light cured as recommended by the manufacturer; the brackets were then placed, and the adhesive was light cured. In group 3, Pro Seal was applied, the bracket with the adhesive was placed on the tooth, and both Pro Seal and the adhesive were cured simultaneously. The purpose of this modification was to reduce one of the steps during the bonding procedure. A shear force was applied at the bracket-tooth interface using a Zwick Universal Test Machine. The results of the analysis of variance (F-ratio = 1.35) indicated that the shear bond strengths of the three groups were not significantly different (P = .267) from each other. The mean shear bond strength of the control group was 4.9+/-2.1 MPa. The mean shear bond strength for teeth coated with Pro Seal and light cured followed by application and light curing of the adhesive was 4.8+/-2.3 MPa, and the mean for the teeth coated with Pro Seal and then bracket placed followed by simultaneous light curing of the sealant and the adhesive was 4.0+/-1.5 MPa. PMID- 15825791 TI - Acidic soft drinks effects on the shear bond strength of orthodontic brackets and a scanning electron microscopy evaluation of the enamel. AB - This study investigated the effects of acidic soft drinks on the resistance of metal brackets to shear forces in vitro and in vivo. Thirty noncarious maxillary premolar teeth, scheduled for extraction for orthodontic purposes, were used in the in vivo group. Thirty other noncarious maxillary premolar teeth, already extracted for orthodontic purposes, were used in the in vitro group. The teeth in both groups were divided equally in three subgroups, ie, the Coca-Cola, Sprite, and control subgroups. Brackets were bonded using conventional methods. Teeth in the in vivo group were rinsed with the acidic drink three times for five minutes daily and extracted after three months. Teeth in the in vitro group were kept in the acidic drink for five minutes on three equal time intervals within 24 hours. The brackets from both groups were subjected to shearing forces using a Universal test machine. After the shearing tests, a scanning electron microscope was used to determine the amount and the localization of erosion. The results indicated that both acidic soft drink subgroups had a reduced debonding resistance in vivo and in vitro compared with their control subgroups. No statistical difference in debonding resistance was found between the in vivo and in vitro groups. Areas of defect due to erosion were observed on the enamel surface around the brackets in both the in vitro and in vivo groups. Acidic soft drinks such as Coca-Cola and Sprite have a negative effect on bracket retention against shearing forces and enamel erosion. PMID- 15825792 TI - Adhesive thickness effects on the bond strength of a light-cured resin-modified glass ionomer cement. AB - These in vitro studies investigated the effect of adhesive thickness on the tensile and shear bond strength of a light-cured, resin-modified glass-ionomer cement (FO). A light-cured conventional composite resin (CO) was used as the control material. Mesh-based metal brackets were bonded to extracted human premolars using FO and CO. The adhesive thickness was controlled by a special device and 0, 0.25, and 0.5 mm thicknesses were tested for both bonding agents. All bonded specimens were stored in distilled water at 37 degrees C for 48 hours and thermocycled between 5 degrees C and 55 degrees C for 200 cycles before testing. Analysis of variance showed that bond strength was significantly affected by the adhesive thickness (P < .001) and type of adhesive (P = .001). There were statistically significant differences between the mean bond strengths of the groups at the P < .05 level of significance. For all adhesive thicknesses, CO had higher bond strength values than those of FO in both test modes. The bond strength values were also analyzed using a Weibull analysis, which showed the most favorable adhesive thickness, and the 5% and 90% probabilities of failures was 0.25 mm in the FO groups. Bracket-adhesive interface failure was predominant for all groups in tensile testing, but enamel-adhesive interface failures increased with increased adhesive thickness in shear testing for the FO. This study suggests that adhesive thickness under a bracket could be particularly important when using a FO in direct bonding. PMID- 15825793 TI - Bonding durability of using self-etching primer with 4-META/ MMA-TBB resin cement to bond orthodontic brackets. AB - This study determines the bonding durability when a self-etching primer is used with Superbond C&B (a 4-methacryloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butyl borane resin) to bond orthodontic brackets to enamel. Thermocycling test was used to assess bonding durability. Metal brackets were bonded to the phosphoric acid-etched or Megabond self-etching primer-treated enamel surface of human premolars using Superbond C&B. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours or after 2000 or 5000 cycles of thermocycling between 5 degrees C and 55 degrees C. Data were analyzed using 2-way analysis of variance and Fisher's protected least significant difference test for multiple comparisons. There was no significant difference in shear bond strength between phosphoric acid and Megabond self etching primer treatment before the thermocycling test. After 2000 and 5000 thermal cycles, significant decreases in shear bond strength were observed with phosphoric acid etching. On the contrary, no significant differences in shear bond strength were observed after 2000 and 5000 thermal cycles with Megabond self etching primer. The adhesive remnant indices were not significantly different between phosphoric acid etching and Megabond self-etching primer treatment either before or after thermal cycles. This study suggested that when used with Superbond C&B in bonding orthodontic brackets, Megabond self-etching primer is superior to phosphoric acid as an enamel preparation agent in providing durable bond strength. PMID- 15825794 TI - Unusual orthodontic correction of bilateral maxillary canine-first premolar transposition. AB - Tooth transposition is a subject that intrigues orthodontists because of the associated treatment planning. Approximately 0.3-0.4% of the population has this type of tooth disharmony, and in the literature, most authors are in disagreement about the treatment approach. In this article, a case is presented of bilaterally maxillary canine-first premolar transposition associated with bilaterally upper lateral incisor agenesis treated in a very unusual way. The transposed teeth were orthodontically reversed to their normal sequence and the missing lateral incisor spaces closed. We choose this approach because once the decision was made to close the upper lateral incisor agenesis spaces, it was inadequate to position the upper first premolars in contact with the central incisors. PMID- 15825795 TI - Nonsurgical treatment of adult open bite using edgewise appliance combined with high-pull headgear and class III elastics. AB - This case report describes the effect of a combination of high-pull headgear and Class III elastics on the nonsurgical treatment of an adult open bite. The 19 year 1-month-old Japanese female presented with the anterior open bite of 4.0 mm and mild crowding. She had a skeletal Class II but a Class III molar relationship due to a severe proclination of the mandibular dental arch. Unilateral congenital missing premolars caused a discrepancy between the facial and dental midline. After extraction of two premolars and the impacted mandibular third molars, nonsurgical therapy was performed using the standard edgewise appliance combined with a high-pull headgear and Class III elastics. The successful treatment outcome and stability of the final occlusion indicates that a combination of high pull headgear and Class III elastics is one of the effective devices in the nonsurgical treatment of open bite and, is especially helpful in uprighting the mandibular dental arch. PMID- 15825797 TI - A new era for the ABO! PMID- 15825796 TI - Maxillary canine transposition to incisor site: a rare condition. AB - Tooth transposition is defined as positional interchange of 2 adjacent teeth within the same quadrant. The incidence has been reported as about 0.4%, and the maxillary permanent canine has been reported as the tooth most frequently involved. Several etiologic factors like genetics, interchange in position of the developing tooth buds, trauma, mechanical interferences, and early loss of incisors have been associated with tooth transposition. Articles in the literature report various kinds of transpositions, but there are only a few reported cases of transposition of the maxillary canine to central incisor site. This kind of transposition has been suggested as resulting primarily by opportunistic canine migration into the space left from an early lost central incisor. However, there are only 2 long-term roentgenographic follow-up studies documenting maxillary canine migration and substantiating this hypothesis. This report shows 2 cases of this rare condition, with 1 of them documented with long term records. PMID- 15825798 TI - Physical examinations for young athletes. PMID- 15825799 TI - Bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease: how safe is eating beef? AB - Cases of bovine spongiform encephalopathy (BSE, mad cow disease) have been found in North American cattle. Its human counterpart, called variant Creutzfeldt-Jakob disease (variant CJD), is rare but seems to be linked to eating diseased beef. Many questions remain about these diseases, such as why young people seem at greater risk of variant CJD. Also, are some people more genetically at risk for acquiring variant CJD than others? PMID- 15825800 TI - Liver biopsy 2005: when and how? AB - Clinical imaging and serologic testing are increasingly replacing biopsy for diagnosing hepatic diseases. However, more biopsies are being done to stage and grade hepatitis C and fatty liver disease, to diagnose space-occupying lesions (typically with fine-needle aspiration biopsy), and to assess response to therapy. If biopsy is planned, it is important to evaluate its indications and risks and, if other physicians are involved, who is responsible for what. PMID- 15825801 TI - The impact of stress urinary incontinence on sexual activity in women. AB - In women, stress urinary incontinence is a common problem that may lead to sexual dysfunction. We review the epidemiological data, the pathophysiology, and the risk factors for these two "closet" disorders, how they are related, how we can get patients to talk about them, and how the treatment of stress urinary incontinence may affect sexual dysfunction. PMID- 15825802 TI - The emergence of methicillin-resistant Staphylococcus aureus in the community. AB - Infections with methicillin-resistant Staphylococcus aureus (MRSA), long endemic in hospitals and nursing homes, are now being reported in the community as well. While we await further epidemiological and microbiological study of this emerging pathogen, current clinical practice requires a reconsideration of the empiric use of beta-lactam agents for the seriously ill patient with a gram-positive infection. PMID- 15825803 TI - Antiphospholipid antibody syndrome: recent findings on managing this challenging condition. AB - Patients with antiphospholipid syndrome can suffer recurrent thrombosis that is very difficult to manage. Recent research has demonstrated that high-intensity anticoagulation is not superior to standard therapy. Basic questions about the nature and treatment of this syndrome remain unanswered. PMID- 15825804 TI - Do hip fractures need to be repaired within 24 hours of injury? PMID- 15825805 TI - Cholesterol guidelines update: more aggressive therapy for higher-risk patients. AB - The 2004 update to the National Cholesterol Education Program guidelines goes farther than the 2001 version in suggesting an optional low-density lipoprotein cholesterol (LDL-C) goal of less than 70 mg/dL for patients at "very high risk." It recommends starting both diet and drug therapy in all patients at high or very high risk whose LDL-C level is above the goal level, with the goal of reducing LDL-C by 30% to 40%. These more aggressive guidelines are based on results of five clinical studies published since 2001. PMID- 15825806 TI - Courting doctors through portals. Hospital looks for advantage as first in market. PMID- 15825807 TI - Valley Health System shifts Web strategy, to market its doctors. PMID- 15825808 TI - Hospital sites plan their moves to more strategic roles: survey asks how, when, why. PMID- 15825809 TI - Dairy intake, obesity, and metabolic health in children and adolescents: knowledge and gaps. AB - There is an urgent need to identify nutrition-related risk factors for obesity and the metabolic syndrome, because the prevalence of these conditions continues to rise among children and adolescents. While some studies suggest that dairy and calcium intake may attenuate obesity and the metabolic syndrome, others do not support these findings. In addition, very little research has been done in children and adolescents, especially in minority youth, who are at the greatest risk for obesity and metabolic dysfunctions. Longitudinal studies examining the role of dairy intake in relation to changes in body composition and metabolic profiles during growth are also critically needed. Of the studies conducted thus far, part of the discrepancy in findings may be due to the uncertainty over whether the effect of dairy intake is independent of energy intake or other eating pattern variables. Further, there is no consensus on how to qualify (i.e., which foods) or quantify (i.e., which cutoffs and/or units) dairy consumption. The widespread problem of implausible dietary reporting in observational studies and the lack of compliance monitoring in intervention trials may also contribute to inconsistent findings. Given the lack of consensus on the effect of dairy, particularly in children and adolescents, more research is warranted before any recommendations can be made on dietary guidelines, policies, and interventions. PMID- 15825810 TI - Biological role of vitamin C in keratinocytes. AB - Epidemiological studies have suggested an association between vitamin C (and other antioxidant vitamins) and cancer risk. However, the mechanisms accounting for prevention have not been extensively investigated. In skin, vitamin C (ascorbic acid) exerts different biological roles, including photoprotective effects and participation in collagen synthesis. This paper reports new findings about additional functions of the vitamin. Vitamin C counteracts oxidative stress via transcriptional and post-translational mechanisms; this modulation may interfere with the activity of redox-sensitive transcription factors, commitment to differentiation or cell cycle arrest, and apoptosis in response to DNA damage. All of these vitamin C-mediated responses might be important in different cell types, allowing for the maintenance of body homeostasis. PMID- 15825811 TI - Influence of vitamin K on anticoagulant therapy depends on vitamin K status and the source and chemical forms of vitamin K. AB - Warfarin therapy requires close monitoring to avoid excessive bleeding and to maintain the effective therapeutic concentration assessed with the internationalized ratio (INR). High vitamin K intake can decrease the therapeutic effectiveness of warfarin, while poor vitamin K status appears to increase the sensitivity to small changes in vitamin K intake, especially from supplements. Very large amounts of vitamin K from a single meal with vegetables (400 g of vegetables with 700 to 1500 microg of vitamin K1) can measurably change INR, but occasional typical servings (<100 g) would probably have little lasting impact on INR. Warfarin requirements may change in those altering their intake of dark green vegetables. The 2005 Dietary Guidelines for Americans recommends 3 cups/week of dark-green vegetables, which contain about 100 to 570 microg/serving of vitamin K1. Less well-known sources and chemical forms of vitamin K, such as MK-7 in natto (a fermented Japanese product), also measurably influence INR. Additional research is needed in warfarin-treated patients to fully quantify the interactions among various sources and chemical forms of vitamin K, age, genotype, and other factors. PMID- 15825812 TI - Function of the protein RPE65 in the visual cycle. AB - A protein called RPE65 performs a key role in the trans-cis isomerization of retinol in the retinal pigment epithelium of the eye. The palmitoylation of RPE65 serves to switch off the visual cycle in darkness and to switch it on in the light. PMID- 15825813 TI - Taking control of your job. PMID- 15825814 TI - Tools for novice health care clinical administrators. AB - With the face of health care changing at a blistering pace due to shrinking resources, advanced technology, and growing consumer demands, organizations, for profit and nonprofit alike, have responded by flattening organizational hierarchies. Consequently, it is not an uncommon practice to promote "successful technical experts" to clinical administrator positions. Ill-prepared, rookie clinical administrators are often left struggling to manage an organization with little experience and training. Fortunately, there is help. This article will identify and apply internal and external tools that newly promoted clinical administrators can use to hurdle potential pitfalls. PMID- 15825815 TI - Social marketing as a tool to improve behavioral health services for underserved populations in transition countries. AB - Human development efforts continue to change the world and improve quality of life for humans. Without the struggle and drive to contemplate new ideas to improve society, the global community would be in a constant state of oppression. Although cultures and norms change as international boundaries are crossed, the universal goal is to improve standards of living to include behavioral health services for underserved populations. In recent times, pioneers and community groups have used social marketing as an instrument to change public perceptions and behaviors within societies. These efforts have transformed nations in the acceptance and understanding of community health and rehabilitation, education, service, and human rights. This article examines the justification for utilization of the concepts and tools of social marketing to bring about proactive behavior modification among segments of underserved populations. A section of this article provides an overview of the basics of social marketing for the benefit of makers of health policy in transition countries. Finally, the case of 2 underserved population segments in the Republic of Slovakia, a new member of the European Union (former socialist block member), is examined for possible implementation. PMID- 15825816 TI - Hospital restructuring stressors, support, and nursing staff perceptions of unit functioning. AB - Hospital restructuring and downsizing have taken place in most developed countries during the past 10 years. A small but growing body of research findings has identified aspects of these changes that serve as sources of stress for nursing staff during these transitions as well as organizational initiatives that facilitate those transitions. This study reports results from a longitudinal study of hospital restructuring and downsizing on nursing staff perceptions of hospital functioning. Data were collected in November 1996 and again in November 1999 from hospital-based nursing staff using questionnaires. The findings showed moderate but statistically significant relationships between restructuring stressors and organizational support in 1996 and perceptions of hospital functioning in 1999. Nursing staff reporting higher levels of restructuring stressors and lower hospital support in 1996 indicated more negative perceptions of unit and hospital functioning in 1999. PMID- 15825817 TI - Hospital nurses' intentions to remain: exploring a northern context. AB - Retaining nurses is of significant concern to all hospitals but even more of a concern to northern and rural hospital managers. This study provides insights into factors related to nurses' intentions to remain. A sample of 122 nurses from 13 northern hospitals in Western Canada participated in the study. The nurses completed questionnaires and participated in structured interviews. A model was proposed which suggested that work experiences (job and decision latitude, feedback, perceptions of how viewed and treated by others, fairness of policies, and safety of the job environment) would be related to job satisfaction and then affective commitment. Age and tenure, and ties to the community were proposed as predictors of continuance commitment. Both affective and continuance commitments were expected to be related to intention to remain in the hospital. The model was partially supported by regression analyses. Work experiences predicted job satisfaction and affective commitment. Affective commitment, continuance commitment, and ties to the community are related to nurses' intentions to remain. Supplemental analyses indicated that the strongest relationships were found for management's views and treatment of nurses, knowledge and ability utilization, safe environment, and fairness of organizational policies. PMID- 15825818 TI - Passion in today's health care leaders. AB - Passion in today's health care leaders is essential as health care organizations face increasing demands for survival. Leaders in health care have been educated, selected, promoted, and retained based on their analytical and creativity skills. Today's health care leaders must also have emotional intelligence. Emotional intelligence is primal for passion. Emotional intelligence, which leads to passion, is crucial to the survivability of today's health care organizations. In order for health care organizations to go from good to great, the leader must inspire followers through passion. This article encourages health care leaders to gain awareness of emotional intelligence and to use emotional intelligence as part of their leadership to inspire passion. Through passion, leaders and followers become more motivated to accomplish the health care mission of serving others. PMID- 15825819 TI - Using a multipronged approach to implement organizational performance improvements. AB - Efforts to improve organizational performance sometimes fail. One way to increase the likelihood of a successful implementation is to adopt a multipronged approach. This article describes a multipronged approach for change management and performance improvement. The approach combines a systems implementation process with sensitivity to the human side of change. Tactics to defuse resistance are also discussed. The approach assists leaders assigned to coordinate performance improvements. PMID- 15825820 TI - Conflict management in public university hospitals in Turkey: a pilot study. AB - By nature, hospitals are extremely complex organizations, combining many different professional groups within an intricate administrative structure. Conflicts therefore expectedly arise between individuals, groups, and departments. It is in the interest of health care administrators to periodically assess the major factors giving rise to these conflicts. In this study, a questionnaire designed to measure sources of conflict in the workplace was completed by 204 staff members at Gazi University Hospital. Of the participants, 30.9% were physicians, and 12.5% were administrators at various levels; 61.5% were female, and 38.5% were male. In terms of work experience, 52.6% of participants had worked less than 5 years at the hospital. The results of the study show that educational differences among the hospital staff were a major barrier to good communication and information flow between groups. Professionals in the same specialties experienced fewer conflicts. Another source of conflict was that resource allocation was considered unfair across departments. Although the hospital management provided an ombudsman for staff concerns, staff rarely resorted to the ombudsman because of the stigma associated with complaining. A lack of opportunity for career advancement was mentioned by 52% of the participants as a source of conflict. At present, job performance and rewards are not closely related in public university hospitals in Turkey because promotions and pay raises are strictly limited by law. Bureaucracy was also perceived to be a source of conflict, with 48.4% of participants saying that their performance was less than optimal because of the presence of multiple supervisors. This pilot study suggests that in Turkey, legislative reform is needed to give public university hospitals more flexibility regarding work incentives, open-door policies at the administrative level, and social interactions to improve teamwork among hospital staff. PMID- 15825821 TI - Incorporating ethics into your comprehensive organizational plan. AB - Today's health care executives find their organizations facing internal and external environments that are behaving in chaotic and unpredictable ways. From inadequate staffing and an increase in clinical errors to outdated risk management procedures and increased competition for scare reimbursements, these health care managers find themselves making decisions without being fully informed of the ethical ramifications of these decisions. A 6-part Comprehensive Organizational Plan is presented that helps the health care decision maker better understand the key success factors for the organization. The Comprehensive Organizational Plan is an overall plan that is intended to protect and serve your organization. The 6 plans in the Comprehensive Organizational Plan cover the following areas: competition, facilities, finances, human resources, information management, and marketing. The comprehensive organizational plan includes an overlay of the ethical considerations for each part of the plan. PMID- 15825822 TI - A critical examination of formal and informal mentoring among nurses. AB - This article focuses on how mentoring among nurses can help them to better serve their profession and health care organizations to more successfully achieve their goals. Specifically, it addresses how formal and informal mentoring can yield benefits to both the nursing profession and health care organizations. It presents some advantages and disadvantages of formal and informal mentoring. Finally, it delineates some important considerations that should be carefully analyzed before establishing a formal program. PMID- 15825823 TI - The manager and oral presentations: from conference room to convention hall. AB - Public speaking is a skill that can prove to be of considerable value to health care managers, yet a great many managers have not been trained as speakers and thus tend to avoid speaking situations or approach them with fear and doubt. However, any working manager can become an effective speaker through conscious effort to do so. The keys to developing one's ability to deliver effective oral presentations are preparation and practice. Preparation includes knowing one's subject and one's audience, appropriately organizing the material, learning how to present information according to what one wishes to convey or accomplish, learning how to utilize visual aids, and working to improve one's manner of using language in a public setting. The same guidelines apply whether the manager is speaking to a small audience in a conference room or a large audience at a conference or convention. Beyond following the guidelines for appropriate preparation and delivery, regardless of scope of presentation and size of audience, success and a degree of comfort with public speaking come with practice, practice, practice. PMID- 15825824 TI - A manager asks: Working and coping with physicians. PMID- 15825825 TI - The effect of alpha-linolenic acid and linoleic acid on the growth and development of formula-fed infants: a systematic review and meta-analysis of randomized controlled trials. AB - This systematic review and meta-analysis aimed to evaluate the effect of modifying 18-carbon PUFA [18-C PUFA: alpha-linolenic acid (ALA, 18:3n-3) and linoleic acid (LA, 18:2n-6)] in the diets of term and preterm infants on DHA (22:6n-3) status, growth, and developmental outcomes. Only randomized controlled trials (RCT) involving formula-fed term and preterm infants, in which the 18-C PUFA composition of the formula was changed and growth or developmental outcomes were measured, were included. Differences were presented as control (standard formula) and treatment (18-C PUFA-supplemented formula). Primary analyses for term infants were 4 and 12 mon and for preterm infants 37-42 and 57 wk postmenstrual age. Five RCT involving term infants and three RCT involving preterm infants were included in the systematic review. Infants fed ALA supplemented formula had significantly higher plasma and erythrocyte phospholipid DHA levels than control infants. There was no effect of ALA supplementation on the growth of preterm infants. In term infants, ALA supplementation was associated with increased weight and length at 12 mon, which was at least 4 mon after the end of dietary intervention. Developmental indices of term infants did not differ between groups. There was a transient improvement in the retinal function of preterm infants fed ALA-supplemented diets compared with controls. The findings suggest that ALA-supplemented diets improve the DHA status of infants. Further studies are needed to provide convincing evidence regarding the effects of ALA supplementation of formula on infant growth and development. PMID- 15825826 TI - Highly unsaturated fatty acid synthesis in vertebrates: new insights with the cloning and characterization of a delta6 desaturase of Atlantic salmon. AB - Fish are an important source of the n-3 highly unsaturated fatty acids (HUFA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids that are crucial to the health of higher vertebrates. The synthesis of HUFA involves enzyme-mediated desaturation, and a delta5 fatty acyl desaturase cDNA has been cloned from Atlantic salmon (Salmo salar) and functionally characterized previously. Here we report cloning and functional characterization of a delta6 fatty acyl desaturase of Atlantic salmon and describe its genomic structure, tissue expression, and nutritional regulation. A salmon genomic library was screened with a salmon delta5 desaturase cDNA and positive recombinant phage isolated and subcloned. The full-length cDNA for the putative fatty acyl desaturase was shown to comprise 2106 bp containing an open reading frame of 1365 bp specifying a protein of 454 amino acids (GenBank accession no. AY458652). The protein sequence included three histidine boxes, two transmembrane regions, and an N-terminal cytochrome b5 domain containing the heme-binding motif HPGG, all of which are characteristic of microsomal fatty acid desaturases. Functional expression showed that this gene possessed predominantly delta6 desaturase activity. Screening and sequence analysis of the genomic DNA of a single fish revealed that the delta6 desaturase gene constituted 13 exons in 7965 bp of genomic DNA. Quantitative real-time PCR assay of gene expression in Atlantic salmon showed that both delta6 and delta5 fatty acyl desaturase genes, and a fatty acyl elongase gene, were highly expressed in intestine, liver, and brain, and less so in kidney, heart, gill, adipose tissue, muscle, and spleen. Furthermore, expression of both delta6 and delta5 fatty acyl desaturase genes in intestine, liver, red muscle, and adipose tissue was higher in salmon fed a diet containing vegetable oil than in fish fed a diet containing fish oil. PMID- 15825827 TI - Molecular evidence that the rate-limiting step for the biosynthesis of arachidonic acid in Mortierella alpina is at the level of an elongase. AB - The oil-producing fungus Mortierella alpina 1S-4 is an industrial strain for arachidonic acid (AA) production. To determine its physiological properties and to clarify the biosynthetic pathways for PUFA, heterologous and homologous gene expression systems were established in this fungus. The first trial was performed with an enhanced green fluorescent protein gene to assess the transformation efficiency for heterologous gene expression. As a result, strong fluorescence was observed in the spores of the obtained transformant, suggesting that the foreign gene was inherited by the spores. The next trial was performed with a homologous PUFA elongase (GLELOp) gene, this enzyme having been reported to catalyze the elongation of GLA (18:3n-6) to dihomo-gamma-linolenic acid (20:3n-6), and to be the rate-limiting step of AA production. The FA composition of the transformant was different from that of the host strain: The GLA content was decreased whereas that of AA was increased. These data support the hypothesis that the GLELOp enzyme plays an important role in PUFA synthesis, and may indicate how to control PUFA biosynthesis. PMID- 15825828 TI - Over-expression of hepatic neutral cytosolic cholesteryl ester hydrolase in mice increases free cholesterol and reduces expression of HMG-CoAR, CYP27, and CYP7A1. AB - Hepatic neutral cytosolic cholesteryl ester hydrolase (hncCEH) is a key enzyme in the regulation of hepatic free cholesterol (FC). In examining the effects of over expression of this enzyme on cholesterol homeostasis, mice were infected with a recombinant adenovirus construct (AdCEH) of the rat hncCEH cDNA driven by the human cytomegalovirus promoter. Cholesteryl esterase and p-nitrophenylcaprylate (PNPC) esterase activities were measured in liver postmitochondrial supernatants at 1, 3, 7, and 11 d after infection with AdCEH or a control virus expressing beta-galactosidase (AdbetaGAL). The PNPC esterase activity of AdCEH mice peaked threefold higher than controls on day 2, declining on subsequent days. In contrast, cholesteryl esterase peaked eightfold higher than controls on day 3, indicating a shift in substrate selectivity of hncCEH. Hepatic FC peaked at 144% of controls, 7 d postinfection. The mRNAs for cholesterol 7alpha-hydroxylase, sterol 27-hydroxylase, and HMG-CoA reductase decreased to 47, 46, and 58% of controls, respectively, on day 7, coinciding with peak FC concentrations. Coinciding with increased cholesteryl esterase activity, hepatic esterified cholesterol dropped precipitously from day 3 onward, to 11% of controls by day 11. Hepatic TAG levels also declined, consistent with the reported TAG lipase activity of hncCEH. These results demonstrate elevation of FC and depletion of cholesteryl esters by over-expression of hncCEH, which were resistant to compensatory responses by other enzymes of cholesterol homeostasis. PMID- 15825829 TI - beta-Oxidation capacity of red and white muscle and liver in Atlantic salmon (Salmo salar L.)--effects of increasing dietary rapeseed oil and olive oil to replace capelin oil. AB - Post-smolt Atlantic salmon (Salmo salar) were fed six diets in which capelin oil was replaced with 0, 25, 50, 75, or 100% rapeseed oil (RO; low-erucic acid) or 50% olive oil (OO). The experimental diets were fed to single groups of Atlantic salmon for 42 wk, whereas the 100% capelin oil (0% RO) diet was fed in duplicate. The beta-oxidation capacity of palmitoyl-CoA was determined, using a method optimized for salmon tissues, at the start of the experiment, after 21 wk (October), and after 42 wk (March) in red and white muscle and in liver. Red muscle showed the highest specific beta-oxidation capacity, but when expressed as total beta-oxidation capacity for the whole tissue, white muscle was the most important tissue for the beta-oxidation of FA. From the initial to the final sampling, the beta-oxidation capacity of white muscle increased significantly, whereas the beta-oxidation capacity in liver decreased significantly. After 22 wk, white muscle exhibited an increased beta-oxidation capacity when the dietary RO content was raised from 25 to 75%, with similar effects in red muscle and liver after 42 wk of feeding. The present results also show that the beta oxidation capacity increased with an increase in fish size. PMID- 15825830 TI - Sulfur-substituted and alpha-methylated fatty acids as peroxisome proliferator activated receptor activators. AB - FA with varying chain lengths and an alpha-methyl group and/or a sulfur in the beta-position were tested as peroxisome proliferator-activated receptor (PPAR)alpha, -delta(beta), and -gamma ligands by transient transfection in COS-1 cells using chimeric receptor expression plasmids, containing cDNAs encoding the ligand-binding domain of PPARalpha, -delta, and -gamma. For PPARalpha, an increasing activation was found with increasing chain length of the sulfur substituted FA up to C14-S acetic acid (tetradecylthioacetic acid = TTA). The derivatives were poor, and nonsignificant, activators of PPARdelta. For PPARgamma, activation increased with increasing chain length up to C16-S acetic acid. A methyl group was introduced in the alpha-position of palmitic acid, TTA, EPA, DHA, cis9,trans11 CLA, and trans10,cis12 CLA. An increased activation of PPARalpha was obtained for the alpha-methyl derivatives compared with the unmethylated FA. This increase also resulted in increased expression of the two PPARalpha target genes acyl-CoA oxidase and liver FA-binding protein for alpha methyl TTA, alpha-methyl EPA, and alpha-methyl DHA. Decreased or altered metabolism of these derivatives in the cells cannot be excluded. In conclusion, saturated FA with sulfur in the beta-position and increasing carbon chain length from C9-S acetic acid to C14-S acetic acid have increasing effects as activators of PPARalpha and -gamma in transfection assays. Furthermore, alpha-methyl FA derivatives of a saturated natural FA (palmitic acid), a sulfur-substituted FA (TTA), and PUFA (EPA, DHA, c9,t11 CLA, and t10,c12 CLA) are stronger PPARalpha activators than the unmethylated compounds. PMID- 15825831 TI - High-fat diets impede the lowering effect of cyclosporine A on rat brain lipids and interact with the expression of apolipoproteins E and J. AB - Cyclosporine A (CsA), a common immunosuppressive agent, produces hyperlipidemia and apolipoprotein profile alterations in plasma as well as neurological and psychiatric complications. In rats, 10 mg CsA/kg/d treatments for 3 wk induce alterations of the electroencephalogram, and of the blood and brain lipids. Using this model, we evaluated whether triacylglycerol (TG)- and lecithin (PC)-enriched diets, reported to decrease epileptic episodes (TG) and to improve memory, could modify the effects of CsA treatment on brain lipids and possibly change apolipoprotein (apo) E and apoJ gene expression. To evaluate this hypothesis, three groups of rats were treated for 3 wk with CsA and received a low-fat, PC, or TG diet. Three other groups were fed the above-mentioned diets and were treated with the CsA solvent. As a control, one group was fed only the low-fat diet. The CsA-mediated decreases in brain cholesterol and PC contents, under a low-fat diet, were eliminated by the TG and PC diets. These high-fat diets induced a global increase in hippocampal transcriptional activity, as revealed by elevated polyadenylated RNA levels. The apoE and apoJ mRNA levels in the cortex and hippocampus of rats receiving the solvent were not statistically different between the TG- and PC-enriched diets but showed important variations compared with the low-fat diet solvent-treated group. A differential effect between the two high-fat diets was observed in the hippocampus, resulting in a significant increase of the apoE to apoJ ratio with the PC diet. The balance between apoE and apoJ is presumed to be important in encephalopathic mechanisms, by its involvement through low levels of brain cholesterol and PC, that might be associated with mental disorders. Our results therefore suggest that diet enrichment with polyunsaturated fat might be beneficial during CsA therapy. However, if the high levels in PC used here are more beneficial on CsA peripheral side effects than similar enrichment in TG, this does not seem to be the case in the brain. Thus, lower levels in PC should be tested. PMID- 15825832 TI - Restoration of depressed prostanoid-induced ileal contraction in spontaneously hypertensive rats by dietary fish oil. AB - We have reported that dietary fish oil (FO) rich in n-3 PUFA modulates gut contractility. It was further demonstrated that the gut of spontaneously hypertensive rats (SHR) has a depressed contractility response to prostaglandins (PG) compared with normotensive Wistar-Kyoto (WKY) rats. We investigated whether feeding diets supplemented with n-3 PUFA increased gut contractility and restored the depressed prostanoid response in SHR gut. Thirteen-week-old SHR were fed diets containing fat at 5 g/100 g as coconut oil (CO), lard, canola oil containing 10% (w/w) n-3 FA as alpha-linolenic acid (1 8:3n-3), or FO (as HiDHA, 22:6n-3) for 12 wk. A control WKY group was fed 5 g/100 g CO in the diet. As confirmed, the SHR CO group had a significantly lower gut response to PGE2 and PGF2alpha compared with the WKY CO group. Feeding FO increased the maximal contraction response to acetylcholine in the ileum compared with all diets and in the colon compared with lard, and restored the depressed response to PGE2 and PGF2alpha in the ileum but not the colon of SHR. FO feeding also led to a significant increase in gut total phospholipid n-3 PUFA as DHA (22:6n-3) with lower n-6 PUFA as arachidonic acid (20:4n-6). Canola feeding led to a small increase in ileal EPA (20:5n-3) and DHA and in colonic DHA without affecting contractility. However, there was no change in ileal membrane muscarinic binding properties due to FO feeding. This report confirms that dietary FO increases muscarinic- and eicosanoid receptor-induced contractility in ileum and that the depressed prostanoid response in SHR ileum, but not colon, is restored by tissue incorporation of DHA as the active nutrient. PMID- 15825833 TI - Dietary n-3 polyunsaturated fatty acids increase T-lymphocyte phospholipid mass and acyl-CoA binding protein expression. AB - Dietary flaxseed oil, which is enriched in alpha-linolenic acid, and fish oil, which is enriched in EPA and DHA, possess anti-inflammatory properties when compared with safflower oil, which is enriched in linoleic acid. The influence of flaxseed oil and fish oil feeding on lipid metabolism in T-lymphocytes is currently unknown. This study directly compared the effects of feeding safflower oil, flaxseed oil, and fish oil for 8 wk on splenic T-lymphocyte proliferation, phospholipid mass, and acyl-CoA binding protein expression in the rat. The data show that both flaxseed oil and fish oil increased acyl-CoA binding protein expression and phosphatidic acid mass in unstimulated T-lymphocytes when compared with safflower oil feeding. Fish oil feeding increased cardiolipin mass, whereas flaxseed oil had no effect. After stimulation, flaxseed oil and fish oil blunted T-lymphocyte interleukin-2 production and subsequent proliferation, which was associated with the lack of increased acyl-CoA binding protein expression. The results reported show evidence for a novel mechanism by which dietary flaxseed oil and fish oil suppress T-lymphocyte proliferation via changes in acyl-CoA binding protein expression and phospholipid mass. PMID- 15825834 TI - Milk fat synthesis is unaffected by abomasal infusion of the conjugated diene 18:3 isomers cis-6,trans-10, cis-12 and cis-6,trans-8,cis-12. AB - It has been previously established that trans-10,cis-12 CLA is a potent inhibitor of milk fat synthesis. Although the mechanism of this action is not completely understood, it has been speculated that eicosanoid-like metabolites of this isomer formed by the activity of tissue desaturases may be responsible for its activity. The objective of this study was to investigate the effects of an enrichment containing an 18:3 conjugated diene, produced in the metabolism of trans-10,cis-12 CLA, on milk fat synthesis. Three rumen-fistulated Holstein cows (210+/-8 d in milk) were randomly assigned in a 3 x 3 Latin square experiment. Treatments were (i) control, (ii) trans-10,cis-12 CLA supplement (2.1 g/d; positive control), (iii) enrichment providing two conjugated diene 18:3 isomers (2.6 g/d of cis-6,trans-10,cis-12 and 4.0 g/d of cis-6,trans-8,cis-12) and trans 10,cis-12 CLA (2.1 g/d). Treatments were abomasally infused for 5 d at 4-h intervals, and there was a 7-d interval between periods. Milk yield, dry matter intake, and milk protein yield were unaffected by treatments. In contrast, the trans-10,cis-12 CLA supplement reduced milk fat yield by 27%, whereas the supplement enriched with conjugated diene 18:3 isomers (treatment iii) had no effect on milk fat yield beyond that attributable to its trans-10,cis-12 CLA content. The transfer efficiency of trans-10,cis-12 CLA into milk fat was 25 and 24% for treatments ii and iii, respectively. At the same time, the abomasally infused conjugated diene 18:3 isomers were transferred to milk fat with an efficiency of 33 and 41% for cis-6,trans-10,cis-12 and cis-6,trans-8,cis-12 18:3, respectively. Overall, short-term abomasal infusion of the conjugated diene 18:3 isomers had no effect on milk fat synthesis, thereby offering no support for an involvement of metabolites of trans-10,cis-12 CLA in the regulation of milk fat synthesis. PMID- 15825835 TI - GC-MS structural characterization of fatty acids from marine aerobic anoxygenic phototrophic bacteria. AB - The FA composition of 12 strains of marine aerobic anoxygenic phototrophic bacteria belonging to the genera Erythrobacter, Roseobacter, and Citromicrobium was investigated. GC-MS analyses of different types of derivatives were performed to determine the structures of the main FA present in these organisms. All the analyzed strains contained the relatively rare 11-methyloctadec-12-enoic acid, and three contained 12-methyl-octadec-11-enoic acid, which has apparently never been reported before. High amounts of the very unusual octadeca-5,11-dienoic acid were present in 9 of the 12 strains analyzed. A FA containing a furan ring was detected in three strains. Analytical data indicated that this FA was 10,13-epoxy 11-methyloctadeca-10,12-dienoic acid. A very interesting enzymatic peroxidation of the allylic carbon 10 of cis-vaccenic acid was observed in three strains. Deuterium labeling and GC-MS analyses enabled us to demonstrate that this enzymatic process involves the initial dioxygenase-mediated formation of 10 hydroperoxyoctadec-11(cis)-enoic acid, which is then isomerized to 10 hydroperoxyoctadec-11(trans)-enoic acid and converted to the corresponding hydroxyacids and oxoacids. Different biosynthetic pathways were proposed for these different compounds. PMID- 15825836 TI - Preparation and fractionation of conjugated trienes from alpha-linolenic acid and their growth-inhibitory effects on human tumor cells and fibroblasts. AB - Conjugated alpha-linolenic acid (CLnA) was prepared from alpha-linolenic acid (9,12,15-18:3n-3, LnA) by alkaline treatment; we fractionated CLnA into three peaks by reversed-phase column-HPLC as evidenced by monitoring absorbance at 205, 235, and 268 nm. Peak I was a conjugated dienoic FA derived from LnA, whereas Peaks II and III were conjugated trienoic LnA. Proton NMR analysis showed that Peak III consisted of the all-trans isomer. The methylated Peak III was further divided into five peaks (Peaks IV-VIII) by silver ion column-HPLC. Peak V, a major constituent in the Peak III fraction, was identified as conjugated 10t,12t,14t-LnA by GC-EIMS and 1H NMR analysis. Peaks III and V, which consisted of conjugated all-trans trienoic LnA, had stronger growth-inhibitory effects on human tumor cell lines than the other collected peaks and strongly induced lipid peroxidation as compared with Peaks I, II, and LnA. We propose that conjugated all-trans trienoic FA have the strongest growth-inhibitory effect among the conjugated trienoic acids and conjugated dienoic acids produced by alkaline treatment of alpha-LnA, and that this effect is mediated by lipid peroxidation. PMID- 15825837 TI - Bioengineering and translational research. PMID- 15825838 TI - When money wasn't king. PMID- 15825839 TI - Teaching entrepreneurship in senior design courses. PMID- 15825840 TI - Cardiac resynchronization therapy. PMID- 15825841 TI - Complementary and alternative medicine in the technology age. Applying bioengineering to understand and evaluate the efficacy of age-old remedies. PMID- 15825842 TI - Bridging conventional medicine and complementary and alternative medicine. BME's pivotal role in the growth of alternative and integrative medicine. PMID- 15825843 TI - Functional magnetic resonance imaging of real and sham acupuncture. Noninvasively measuring cortical activation from acupuncture. PMID- 15825844 TI - Using ultrasound to understand acupuncture. Acupuncture needle manipulation and its effect on connective tissue. PMID- 15825845 TI - The application of fMRI to basic experiments in acupuncture. The effects of stimulus points and content on cerebral activities and responses. PMID- 15825846 TI - Sympathetic nervous system activity in stress and biofeedback relaxation. Monitoring SNS activity with the photoplethysmographic-wave envelope and temperature-variability signals. PMID- 15825847 TI - Massage therapy in the treatment of lymphedema. Rationale, results, and applications. AB - The ongoing NCCAM-supported experimental and clinical translational approaches should shed light not only on the physiologic mechanisms underlying the benefits of massage therapy but could also, if successful in defined populations of patients, have a substantial impact by providing a simpler, more cost-effective LE treatment alternative worldwide. PMID- 15825848 TI - Using a healing robot for the scientific study of shirodhara. Altered states of consciousness and decreased anxiety through Indian dripping oil treatments. PMID- 15825849 TI - Molecular prospecting for drugs from the sea. Isolating therapeutic peptides and proteins from cone snail venom. PMID- 15825850 TI - User-interactive registration of bone with A-mode ultrasound. PMID- 15825851 TI - The informative-capacity phenomenon of drying drops. PMID- 15825852 TI - The Vioxx debacle. PMID- 15825853 TI - Cardiopulmonary resuscitation development. PMID- 15825854 TI - COMAR Technical Information Statement. The IEEE exposure limits for radiofrequency and microwave energy. PMID- 15825855 TI - Extinction coefficients of hemoglobin for near-infrared spectroscopy of tissue. PMID- 15825856 TI - The challenge of integrating the healthcare enterprise. PMID- 15825857 TI - Pulsatile cerebrospinal fluid dynamics in the human brain. AB - Disturbances of the cerebrospinal fluid (CSF) flow in the brain can lead to hydrocephalus, a condition affecting thousands of people annually in the US. Considerable controversy exists about fluid and pressure dynamics, and about how the brain responds to changes in flow patterns and compression in hydrocephalus. This paper presents a new model based on the first principles of fluid mechanics. This model of fluid-structure interactions predicts flows and pressures throughout the brain's ventricular pathways consistent with both animal intracranial pressure (ICP) measurements and human CINE phase-contrast magnetic resonance imaging data. The computations provide approximations of the tissue deformations of the brain parenchyma. The model also quantifies the pulsatile CSF motion including flow reversal in the aqueduct as well as the changes in ICPs due to brain tissue compression. It does not require the existence of large transmural pressure differences as the force for ventricular expansion. Finally, the new model gives an explanation of communicating hydrocephalus and the phenomenon of asymmetric hydrocephalus. PMID- 15825858 TI - Encoding and decoding target locations with waves in the turtle visual cortex. AB - Visual stimuli elicit waves of activity that propagate across the visual cortex of turtles. An earlier study showed that these waves encode information about the positions of stimuli in visual space. This paper addresses the question of how this information can be decoded from the waves. Windowing techniques were used to temporally localize information contained in the wave. Sliding encoding windows were used to represent waves of activity as low dimensional temporal strands in an appropriate space. Expanding detection window (EDW) or sliding detection window (SDW) techniques were combined with statistical hypothesis testing to discriminate input stimuli. Detection based on an EDW was more reliable than detection based on a SDW. Detection performance improved at a very early stage of the cortical response as the length of the detection window is increased. The property of intrinsic noise was explicitly considered. Assuming that the noise is colored provided a more reliable estimate than did the assumption of a white noise in the cortical output. PMID- 15825859 TI - Comparing generalized and phase synchronization in cardiovascular and cardiorespiratory signals. AB - We made use of multivariate nonlinear analysis methods to study the interdependence between the cardiac interval variability and both the respiratory activity and the systolic pressure in rats. The study was carried out in basal conditions and after the application of different drugs affecting the cardiovascular system. The results showed that there are changes both in the extent and in the directionality of such interdependences because of the drugs. The inhibition of the NO and the parasympathetic blockade changed the cardiovascular coordination, with the latter one also modifying the interdependence between the cardiac interval and the respiratory signal. This suggests that the nonlinear approach might be very helpful to explore the interaction between subsystems of the cardiovascular control system. PMID- 15825860 TI - Mutual information function assesses autonomic information flow of heart rate dynamics at different time scales. AB - The autonomic information flow (AIF) represents the complex communication within the Autonomic Nervous System (ANS). It can be assessed by the mutual information function (MIF) of heart rate fluctuations (HRF). The complexity of HRF is based on several interacting physiological mechanisms operating at different time scales. Therefore one prominent time scale for HRF complexity analysis is not given a priori. The MIF reflects the information flow at different time scales. This approach is defined and evaluated in the present paper. In order to aggregate relevant physiological time scales, the MIF of HRF obtained from eight adult Lewis rats during the awake state, under general anesthesia, with additional vagotomy, and additional betal-adrenergic blockade are investigated. Physiologically relevant measures of the MIF were assessed with regard to the discrimination of these states. A simulation study of a periodically excited pendulum is performed to clarify the influence of the time scale of MIF in comparison to the Kolmogorov Sinai entropy (KSE) of that well defined system. The general relevance of the presented AIF approach was confirmed by comparing mutual information, approximate entropy, and sample entropy at their respective time scales. PMID- 15825861 TI - High-resolution analysis of ambulatory electrocardiograms to detect possible mechanisms of premature ventricular beats. AB - For generations of electrocardiogram (ECG) analysis, the presence of premature ventricular beats (PVBs) has been characterized as a common event in the ECG without regard to the mechanism which has caused the PVB in the first place. At best, the coupling interval with the preceding sinus beat may be noted. This viewpoint persisted throughout the era of automated ECG analysis, as well as influencing the treatment of more life threatening events by PVB suppression strategies alone. This study proposed three hypotheses which would link the PVB to a specific mechanism or milieu. Each of these hypotheses requires significant signal processing of the continuously recorded high resolution ECG. Data are presented which demonstrate that abnormal intra-QRS potentials may be linked to a reentrant mechanism for the PVBs and that many patients have significant changes in these potentials in the sinus beats preceding the PVB. Changes in the characteristics of the repolarization as measured in the T/U wave period were also observed and could be linked to triggered activity mechanism for some PVBs. Finally, the role of subclinical ST segment changes also indicates that low grade ischemia may play a role in modulating either PVB mechanism. The data generated by this study suggest that a new view toward PVB mechanism as measured by ECG characteristics may warrant a more rational approach to renewed interest identifying the malignant PVBs and their eventual clinical management. PMID- 15825862 TI - Methodological principles of T wave alternans analysis: a unified framework. AB - Visible T wave alternans (TWA) in the electrocardiogram (ECG) had been regarded as an infrequent phenomenon during the first 80 years of electrocardiography. Nevertheless, computerized analysis changed this perception. In the last two decades, a variety of techniques for automatic TWA analysis have been proposed. These techniques have allowed researchers to detect nonvisible TWA in a wide variety of clinical and experimental conditions. Such studies have recently shown that TWA is related to cardiac instability and increased arrhythmogenicity. Comparison of TWA analysis methods is a difficult task due to the diversity of approaches. In this paper, we propose a unified framework which holds the existing methods. In the light of this framework, the methodological principles of the published TWA analysis schemes are compared and discussed. This framework may have an important role to develop new approaches to this problem. PMID- 15825863 TI - Automated estimation of the phase between thoracic and abdominal movement signals. AB - This paper presents a new procedure for the automated estimation of the phase relation between thoracic and abdominal breathing signals measured by inductance plethysmography (RIP). This estimation is achieved using linear filters, binary converters and an exclusive-or gate. The filters are designed offline from prior knowledge of the spectrum of subjects' respiration, reducing computational complexity and providing on-line processing capabilities. Some numerical results based on simulated time series and infant respiration data are provided, showing that the new method is less biased than the Pearson correlation method, commonly used for assessment of thoracoabdominal asynchrony. Our method offers further advantages: 1) it works with uncalibrated measurements; 2) it provides quantitative phase estimates with no need to estimate the underlying frequency of the breathing signals; 3) it does not require nonconvex optimization search algorithms; and 4) it is easy to implement and to automate. PMID- 15825864 TI - A technique to track individual motor unit action potentials in surface EMG by monitoring their conduction velocities and amplitudes. AB - The speed of propagation of an action potential along a muscle fiber, its conduction velocity (CV), can be used as an indication of the physiological or pathological state of the muscle fiber membrane. The motor unit action potential (MUAP), the waveform resulting from the spatial and temporal summation of the individual muscle fiber action potentials of that motor unit (MU), propagates with a speed referred to as the motor unit conduction velocity (MUCV). This paper introduces a new algorithm, the MU tracking algorithm, which estimates MUCVs and MUAP amplitudes for individual MUs in a localized MU population using SEMG signals. By tracking these values across time, the electrical activity of the localized MU pool can be monitored. An assessment of the performance of the algorithm has been achieved using simulated SEMG signals. It is concluded that this analysis technique enhances the suitability of SEMG for clinical applications and points toward a future of noninvasive diagnosis and assessment of neuromuscular disorders. PMID- 15825865 TI - Quantifying errors in spectral estimates of HRV due to beat replacement and resampling. AB - Spectral estimates of heart rate variability (HRV) often involve the use of techniques such as the fast Fourier transform (FFT), which require an evenly sampled time series. HRV is calculated from the variations in the beat-to-beat (RR) interval timing of the cardiac cycle which are inherently irregularly spaced in time. In order to produce an evenly sampled time series prior to FFT-based spectral estimation, linear or cubic spline resampling is usually employed. In this paper, by using a realistic artificial RR interval generator, interpolation and resampling is shown to result in consistent over-estimations of the power spectral density (PSD) compared with the theoretical solution. The Lomb-Scargle (LS) periodogram, a more appropriate spectral estimation technique for unevenly sampled time series that uses only the original data, is shown to provide a superior PSD estimate. Ectopy removal or replacement is shown to be essential regardless of the spectral estimation technique. Resampling and phantom beat replacement is shown to decrease the accuracy of PSD estimation, even at low levels of ectopy or artefact. A linear relationship between the frequency of ectopy/artefact and the error (mean and variance) of the PSD estimate is demonstrated. Comparisons of PSD estimation techniques performed on real RR interval data during minimally active segments (sleep) demonstrate that the LS periodogram provides a less noisy spectral estimate of HRV. PMID- 15825866 TI - Comparison of trend detection algorithms in the analysis of physiological time series data. AB - This paper presents a comparative performance analysis of various trend detection methods developed using fuzzy logic, statistical, regression, and wavelet techniques. The main contribution of this paper is the introduction of a new method that uses noise rejection fuzzy clustering to enhance the performance of trend detection methodologies. Furthermore, another contribution of this work is a comparative investigation that produced systematic guidelines for the selection of a proper trend detection method for different application requirements. Examples of representative physiological variables considered in this paper to examine the trend detection algorithms are: 1) blood pressure signals (diastolic and systolic); and 2) heartbeat rate based on RR intervals of electrocardiography signal. Furthermore, synthetic physiological data intentionally contaminated with various types of real-life noise has been generated and used to test the performance of trend detection methods and develop noise-insensitive trend detection algorithms. PMID- 15825867 TI - Elastic registration of biological images using vector-spline regularization. AB - We present an elastic registration algorithm for the alignment of biological images. Our method combines and extends some of the best techniques available in the context of medical imaging. We express the deformation field as a B-spline model, which allows us to deal with a rich variety of deformations. We solve the registration problem by minimizing a pixelwise mean-square distance measure between the target image and the warped source. The problem is further constrained by way of a vector-spline regularization which provides some control over two independent quantities that are intrinsic to the deformation: its divergence, and its curl. Our algorithm is also able to handle soft landmark constraints, which is particularly useful when parts of the images contain very little information or when its repartition is uneven. We provide an optimal analytical solution in the case when only landmarks and smoothness considerations are taken into account. We have applied our approach to perform the elastic registration of images such as electrophoretic gels and fly embryos. The validation of the results by experts has been favorable in all cases. PMID- 15825868 TI - Fluoroscopy-based 3-D reconstruction of femoral bone cement: a new approach for revision total hip replacement. AB - In revision total hip replacement the removal of the distal femoral bone cement can be a time consuming and risky operation due to the difficulty in determining the three-dimensional (3-D) boundary of the cement. We present a new approach to reconstruct the bone cement volume by using just a small number of calibrated multiplanar X-ray images. The modular system design allows the surgeon to react intraoperatively to problems arising during the individual situation. When encountering problems during conventional cement removal, the system can be used on demand to acquire a few calibrated X-ray images. After a semi-automatic segmentation and 3-D reconstruction of the cement with a deformable model, the system guides the surgeon through a free-hand navigated or robot-assisted cement removal. The experimental evaluation using plastic test implants cemented into anatomic specimen of human femoral bone has shown the potential of this method with a maximal error of 1.2 mm (0.5 mm RMS) for the distal cement based on just 4 5 multiplanar X-ray images. A first test of the complete system, comparing the 3 D-reconstruction with a computed tompgraphy data set, confirmed these results with a mean error about 1 mm. PMID- 15825869 TI - Automated extraction of temporal motor activity signals from video recordings of neonatal seizures based on adaptive block matching. AB - This paper presents an automated procedure developed to extract quantitative information from video recordings of neonatal seizures in the form of motor activity signals. This procedure relies on optical flow computation to select anatomical sites located on the infants' body parts. Motor activity signals are extracted by tracking selected anatomical sites during the seizure using adaptive block matching. A block of pixels is tracked throughout a sequence of frames by searching for the most similar block of pixels in subsequent frames; this search is facilitated by employing various update strategies to account for the changing appearance of the block. The proposed procedure is used to extract temporal motor activity signals from video recordings of neonatal seizures and other events not associated with seizures. PMID- 15825870 TI - A system-on-chip digital pH meter for use in a wireless diagnostic capsule. AB - This paper describes the design and implementation of a system-on-chip digital pH meter, for use in a wireless capsule application. The system is organized around an 8-bit microcontroller, designed to be functionally identical to the Motorola 6805. The analog subsystem contains a floating-electrode ISFET, which is fully compatible with a commercial CMOS process. On-chip programmable voltage references and multiplexors permit flexibility with the minimum of external connections. The chip is designed in a modular fashion to facilitate verification and component re-use. The single-chip pH meter can be directly connected to a personal computer, and gives a response of 37 bits/pH, within an operating range of 7 pH units. PMID- 15825871 TI - Temperature dependence of tissue impedivity in electrical impedance tomography of cryosurgery. AB - The temperature-dependent impedivity of rat liver, transverse abdominal muscle and full skin was determined in vitro as a function of frequency across the temperature range 5 degrees C to 37 degrees C and from 100 Hz to 10 kHz. This study was motivated by an increasing interest in using electrical impedance tomography (EIT) for imaging of cryosurgery and a lack of applicable data in the hypothermic range. Using a controlled-temperature impedance analyzer, it was found that as the temperature is reduced the resulting increase in tissue impedivity is more pronounced at low frequencies and that the beta dispersion, resulting from cell membrane polarization, shifts to lower frequencies. With these new data a simple case study of EIT of liver cryosurgery was examined, using a finite-element model incorporating the Pennes bio-heat equation, to determine the impact of this behavior on imaging accuracy. Overestimation of the ice-front position was found to occur if the EIT system ignored the effects of the low-temperature zone surrounding the frozen tissue. This error decreases with increasing blood perfusion and with higher measurement frequencies. PMID- 15825872 TI - Effects of radiofrequency energy on human chondromalacic cartilage: an assessment of insulation material properties. AB - The objective of this study was to establish guidelines for the selection of an insulation material used to surround the electrode of radiofrequency energy (RFE) probes used for thermal chondroplasty. These guidelines were established by identifying which insulation materials resulted in the least amount of chondrocyte death while smoothing the surface of chondromalacic cartilage. RFE causes electrolyte oscillation and molecular friction in the tissue to heat it and subsequently smooth the surface. Material properties investigated included the coefficient of thermal expansion (CTE), thermal conductivity (TC), and volume resistivity (VR). Fresh human chondromalacic cartilage samples of Outerbridge grades II and III were obtained from patients undergoing total knee arthroplasty. Stiffness measurements were taken pretreatment and posttreatment. RFE was applied to a 1-cm2 area for 15 s in a paintbrush treatment pattern. The insulation materials evaluated included Macor (decrease CTE, decrease TC, increase VR; in relation to CTE = 10 x 10(-6)/degrees C at 20 degrees C, TC = 3 W/mK, VR=1 x 10(14) ohm x cm), zirconia toughened alumina (ZTA) and 99.5% alumina (decrease CTE, increase TC, increase VR), aluminum nitride (decrease CTE, increase TC, decrease VR), Teflon (PTFE) (increase CTE, decrease TC, increase VR), partially stabilized zirconia (YTZP) (decrease CTE, decrease TC, decrease VR), and Ultem (increase CTE, decrease TC, decrease VR). There were no significant differences between pretreatment and posttreatment stiffness of the cartilage for any material investigated. Subjectively scored scanning electron microscopy (SEM) images revealed that the surfaces of all samples treated with RFE were relatively smooth with melted fronds. Prototype probes made with Macor, 99.5% alumina, and ZTA had TC < or = 30 W/mol x K and resulted in a mean of 35% less cell death (176+/-56 microm, 130+/-48 microm, and 114+/-33 microm, respectively) than aluminum nitride, PTFE, and YTZP (246+/-68 microm, 231+/-108 microm, and 195+/-89 microm, respectively). Macor, 99.5% alumina, and ZTA prototype probes all had VR > or = 1 x 10(14) ohm x cm and resulted in a mean 37% less cell death than aluminum nitride or YTZP. There was no apparent relationship between CTE and the depth of chondrocyte death. PMID- 15825873 TI - An ultra-low-power programmable analog bionic ear processor. AB - We report a programmable analog bionic ear (cochlear implant) processor in a 1.5 microm BiCMOS technology with a power consumption of 211 microW and 77-dB dynamic range of operation. The 9.58 mm x 9.23 mm processor chip runs on a 2.8 V supply and has a power consumption that is lower than state-of-the-art analog-to-digital (A/D)-then-DSP designs by a factor of 25. It is suitable for use in fully implanted cochlear-implant systems of the future which require decades of operation on a 100-mAh rechargeable battery with a finite number of charge discharge cycles. It may also be used as an ultra-low-power spectrum-analysis front end in portable speech-recognition systems. The power consumption of the processor includes the 100 microW power consumption of a JFET-buffered electret microphone and an associated on-chip microphone front end. An automatic gain control circuit compresses the 77-dB input dynamic range into a narrower internal dynamic range (IDR) of 57 dB at which each of the 16 spectral channels of the processor operate. The output bits of the processor are scanned and reported off chip in a format suitable for continuous-interleaved-sampling stimulation of electrodes. Power-supply-immune biasing circuits ensure robust operation of the processor in the high-RF-noise environment typical of cochlear implant systems. PMID- 15825874 TI - Objective measurement of tactile mislocalization. AB - Stimulating the skin with intensities close to the sensory threshold causes erroneous localization of the site of stimulation. Previous studies using manual methods for applying faint tactile stimuli have shown that localization errors obey a somatotopic principle in which tactile stimuli are preferentially mislocalized to sites adjacent to the stimulated skin region. However, manual testing of mislocalization is time consuming and only partially objective because results depend on the skills of the tester. To improve the testing procedure, an automated apparatus was developed. The procedure adjusted stimulus intensity adaptively during testing to remain near the individual subject's sensory threshold, so that mislocalizations occurred often enough to assess somatotopic organization. The new method was applied to 12 healthy subjects. In each subject, the five digits of the right hand were stimulated singly in random order. Localization errors were distributed preferentially to fingers close to the stimulated finger rather than to distant fingers. The profile of mislocalization differed significantly from that expected on the basis of response bias or guessing behavior. The present results replicate previous findings obtained for manual testing with improved sensitivity and indicate that the new technique is a useful tool for the study of somatosensory processing on a perceptual level. PMID- 15825875 TI - Optimal electrical stimulation modality for cortical esophageal evoked potentials: transmural or intraesophageal? AB - Esophageal electrical stimulation using short and a relatively small number of (200 micros, 0.2 Hz, n = 25) electrical pulses generates a characteristic and well defined cortical evoked potential response (EP). There are two methods of stimulation: either through intraesophageal electrodes or with transmural electrodes. The objective of this paper is to compare EP response, sensations and heart rate variability power spectra elicited by both stimulation modalities in healthy volunteers. Our results suggest that transmural stimulation is more accurately perceived and at lower intensities, produces more reproducible peaks of higher amplitude than during intraesophageal stimulation. During either mode of esophageal stimulation, power within the high-frequency component of the heart rate variability power spectrum is enhanced. PMID- 15825876 TI - Energy-optimal electrical excitation of nerve fibers. AB - We derive, based on an analytical nerve membrane model and optimal control theory of dynamical systems, an energy-optimal stimulation current waveform for electrical excitation of nerve fibers. Optimal stimulation waveforms for nonleaky and leaky membranes are calculated. The case with a leaky membrane is a realistic case. Finally, we compare the waveforms and energies necessary for excitation of a leaky membrane in the case where the stimulation waveform is a square-wave current pulse, and in the case of energy-optimal stimulation. The optimal stimulation waveform is an exponentially rising waveform and necessitates considerably less energy to excite the nerve than a square-wave pulse (especially true for larger pulse durations). The described theoretical results can lead to drastically increased battery lifetime and/or decreased energy transmission requirements for implanted biomedical systems. PMID- 15825877 TI - Noninvasive detection of coronary artery disease by arterio-oscillo-graphy. AB - Coronary artery disease (CAD) causes oscillations in peripheral arteries. Oscillations of the walls of the brachial arteries of 51 patients were recorded [together with the electrocardiogram (ECG)] by an accelerometer at different cuff pressures. By analyzing the energy of the oscillations in the 30-250 Hz band, 16 of 22 patients with CAD and 26 of 29 non-CAD subjects were classified correctly, independent of the ECG, and with no effect of heart murmurs. PMID- 15825878 TI - Improving the accuracy and reliability of motion tracking methods used for extracting temporal motor activity signals from video recordings of neonatal seizures. AB - This paper presents an approach for improving the accuracy and reliability of motion tracking methods developed for video based on block motion models. This approach estimates the displacement of a block of pixels between two successive frames by minimizing an error function defined in terms of the pixel intensities at these frames. The minimization problem is made analytically tractable by approximating the error function using a second-order Taylor expansion. The improved reliability of the proposed method is illustrated by its application in the extraction of temporal motor activity signals from video recordings of neonatal seizures. PMID- 15825879 TI - Comments on "accuracy limitations of chronaxie values". AB - Some additions/corrections are offered to Geddes, 2004. Stimulation is initiated by the second spatial derivative of the voltage along the nerve (activating function) rather than current density. Chronaxie values change with distance from the electrode. Anodic stimulation can excite via anodic break excitation, or via virtual cathodes around the anode. PMID- 15825880 TI - Can squirrel monkeys (Saimiri sciureus) plan for the future? Studies of temporal myopia in food choice. AB - In seven experiments, 2 squirrel monkeys were given choices between arrays of food that varied in the quantity offered. In Experiments 1-5, the monkeys were offered choices between quantities of the same food that varied in a 2:1 ratio. The squirrel monkeys failed to show the temporal myopia effect or a decrease in preference for the larger quantity as the absolute number of food items offered increased. Even when given choices of 8 versus 16 peanuts and 10 versus 20 peanuts, both monkeys significantly preferred the larger quantity. An examination of the monkeys' rates of consumption indicated that 20 peanuts were consumed over a 1- to 2-h period, with eating bouts separated by periods of nonconsumption. In Experiments 6A, 6B, and 7, food was either pilfered or replenished 15 min after an initial choice, so that choice of the smaller quantity led to more total food in the long run. These manipulations caused both monkeys to reduce choice of the larger quantity, relative to baseline choice. The results suggest that squirrel monkeys anticipated the future consequences of their choices. PMID- 15825881 TI - Spatial patterns and memory for locations. AB - Rats obtained food from the tops of vertical poles in a 5 x 5 matrix of locations. On each trial, the baited locations formed one of the two possible exemplars of a checkerboard spatial pattern. During training, locations that had been visited earlier in the trial were indicated by a visual cue. Following training, performance with and without the visual cues was compared. Spatial choices were controlled by the checkerboard spatial pattern. The visual cues enhanced the ability of rats to avoid revisits of locations. However, the visual cues did not enhance control by the spatial pattern, as would be expected if the same spatial memories were involved in avoidance of revisits and coding the location of baited locations. PMID- 15825882 TI - Spontaneous recovery varies inversely with the training-extinction interval. AB - Four experiments found the magnitude of spontaneous recovery after extinction to be greater with a shorter interval between initial conditioning and extinction. Experiments 1 and 2 used a Pavlovian magazine approach procedure with rat subjects, Experiment 3 used an instrumental training procedure with rats, and Experiment 4 used a sign-tracking procedure with pigeons. These results are not anticipated by many accounts of spontaneous recovery that attribute it to the fading of learning that occurred during extinction. PMID- 15825883 TI - Recovery of the rabbit's conditioned nictitating membrane response without direct reinforcement after extinction. AB - Three experiments demonstrated that, following the extinction of an established conditioned stimulus (CSA--e.g., tone), the pairing of a novel, cross-modal stimulus (CSB--e.g., light) with the unconditioned stimulus (US) results in strong recovery of responding to the extinguished CSA. Experiment 1 demonstrated that the recovery of responding to CSA is not the result of US reinstatement but is attributable to pairings of CSB with the US. Experiment 2 demonstrated that the recovery of responding is specific to CSA and is not the result of cross modal generalization. Experiment 3 revealed that a large number of CSB-US pairings in Stage 1 significantly reduced the amount of recovery to CSA during subsequent CSB-US trials. Experiment 3 also provided unexpected evidence of cross modal secondary extinction. The extinction and subsequent recovery of responding seen in the present experiments is discussed with respect to possible contributions from contextual associations, CS processing, US processing, conditioned response expression, and layered excitatory associations. PMID- 15825884 TI - Stable individual differences on developmental tasks in young yellow-crowned parakeets, Cyanoramphus auriceps. AB - We report on stable individual differences in young yellow-crowned parakeets across 38 tasks of cognitive development on three scales involving object permanence, means-end relations, and spatial relations. Stable performance ranks on blocks of tasks emerged after 13 weeks in two groups of sibling parakeets, one hand-reared and the other parent-reared. Examination of subject characteristics, such as hatch order, sex, general activity level, avoidances, latencies, social status, and errors, showed no significant correlations with these performance ranks. Characteristic individual approaches to unstructured play activities (from a fourth scale) were consistent with individual behavioral differences seen on the structured cognitive tasks. PMID- 15825885 TI - Taste + odor interactions in compound aversion conditioning. AB - In three experiments with rats, taste + odor interactions in compound aversion conditioning were investigated. In Experiment 1, two odors (0.02% almond and 0.02% orange) were compared on single-element odor aversions, taste (denatonium) potentiated odor aversions, and potentiated odor aversions following taste extinction. Although no odor differences were seen following single-element conditioning, both types of potentiated orange odor aversions were stronger than their almond odor counterparts. These data show that odors of similar conditionability are differentially potentiated by the same taste. To determine whether these differences were due to unique perceptual representations, the effects of elemental extinction or compound extinction on aversions to the compound were investigated in Experiments 2 and 3. In Experiment 2, orange odor extinction weakened responding to the compound significantly more than taste extinction did. In contrast, almond odor extinction and taste extinction produced similar decrements in responding to the compound in Experiment 3. These results suggest that the perceptual representation of these specific taste + odor compounds are different, and they are discussed in regard to configural and within-compound association accounts of potentiation. PMID- 15825886 TI - A mixed design reveals that glucose moieties facilitate extinction of a conditioned taste aversion in rats. AB - Separate groups of water-deprived rats had four trials with 15-min access to 0.0073 M saccharin, 0.3 M alanine, 0.3 M glucose, 0.1 M maltose, 0.3 M fructose, 0.06 M sucrose, or 0.03 M Polycose. Trials 1-3 were followed by injections of either 0.15 M LiCl (1.33 ml/100 g b.w., i.p.) or saline; Trial 4 (Test) was CS only. Extinction included either 48-h access to water alone or to the appropriate CS, both followed by a 24-h, two-bottle choice of CS and water. This 3-day cycle was repeated five to six times. All rats acquired comparable conditioned taste aversions (CTAs), but extinction rates varied with the test and the taste CS. No CTA extinguished during the two-bottle choices following 2 water days. During one bottle CS exposure, all CTAs extinguished, but the aversion continued longer in the probe two-bottle tests. Intake of glucose moieties recovered rapidly, often in two cycles; the other CSs took four to six cycles. Thus, CTA extinction varies with the nature of the taste CS. PMID- 15825888 TI - Processing of empty and filled time intervals in pigeons. AB - Pigeons were trained initially with 2- and 8-sec empty or filled intervals as sample stimuli. Interval onset and termination was signaled by 1-sec start and stop markers. Following retention and psychophysical testing, both groups were trained with the alternative type of interval, and the tests were repeated. Group empty-first demonstrated a choose-long effect with both empty and filled intervals. Group filled-first demonstrated a weak (and nonsignificant) choose short effect with filled intervals and a robust choose-long effect with empty intervals. Both groups tended to time the markers and to add that duration to the sample duration only on filled-interval trials. Initial training with empty intervals alters the way pigeons process temporal information on filled-interval trials, whereas initial training with filled intervals has little effect on the processing of temporal information on empty-interval trials. PMID- 15825887 TI - Sign-tracking (autoshaping) in rats: a comparison of cocaine and food as unconditioned stimuli. AB - A series of experiments was performed to determine whether sign-tracking would occur in rats with intravenous (i.v.) cocaine as the unconditioned stimulus. In Experiment 1, a retractable lever paired with food produced strong sign-tracking, but a lever paired with one of three doses of i.v. cocaine did not elicit any approach or contact behavior. Experiment 2 demonstrated that doses of cocaine that did not elicit sign-tracking would function as a positive reinforcer for a lever contact operant. In Experiment 3, an artificial consummatory response was added to make the cocaine reinforcement episode more behaviorally comparable to that occasioned by food. Although the rats readily performed this response when it was required to receive cocaine infusions, they still did not contact a lever that signaled the availability of these infusions. It appears that cocaine is different from other positive reinforcers (e.g., food, water, warmth, or intracranial stimulation) in that it will not produce sign-tracking in rats. PMID- 15825889 TI - Acquisition of schedule-induced polydipsia by rats in proximity to upcoming food delivery. AB - Food-deprived rats that receive intermittent delivery of small amounts of food develop excessive drinking--specifically, schedule-induced polydipsia (SIP). A main characteristic of SIP is its occurrence at the beginning of interfood intervals. The purpose of this study was to demonstrate that SIP can be developed toward the end of interfood intervals, in closer proximity to upcoming than to preceding food delivery. In Experiment 1, two groups were exposed to a fixed-time (FT) 30-sec food schedule with water available during the first or the last 15 sec of each interfood interval. Two additional groups, which had access to water throughout, were exposed to FT 30-sec or FT 15-sec schedules of food presentation. The FT 30-sec group with free access to water developed the highest level of intake; similar and intermediate levels were induced in all the remaining groups. In Experiment 2, three groups of rats were exposed to an FT 90 sec food schedule with water available during the first, the second, or the last 30 sec of each interfood interval. One additional group with access to water throughout was exposed to the FT 90-sec schedule of food presentation. The group with free access to water developed a higher level of consumption than did the other groups, but by the end of training none of the four groups showed statistical differences in polydipsic drinking. Results show that adjunctive drinking can be developed in proximity to upcoming food delivery even with long interfood intervals. PMID- 15825890 TI - Effects of runway shift and stay rules on rats' serial pattern learning in the T maze. AB - Rats received three-trial series on a T-maze consisting of extended visually distinct left-black and right-striped side runways. During the first phase of training, when allowed to select baited runways within these series, they predominantly alternated their choices. During the second phase, rats received forced-choice serial pattern training of series consisting of two rewarded (R) trials and one nonrewarded (N) trial in two fixed orders, RRN and RNR. In Experiment 1, the rats in the runway shift rule group always received the second R trial when forced down a runway opposite that on the preceding trial in the series and the N trial when forced down the same runway. The rats in the runway stay rule group always received the second R trial when forced down the same runway and the N trial when forced down the opposite runway. In Experiment 2, each rat was conditionally trained with both runway outcome rules as determined by the central alley lighting and the type of food in the side alleys. The rats took longer to reduce their running speed on the N trial within each sequence under the runway stay rule than under the runway shift rule. They also took longer to acquire serial pattern responding for the RNR than for the RRN series only under the runway stay rule condition. When subsequently reexposed to series of free-choice trials on the final phase, rats maintained spontaneous alternating choice patterns under the runway shift rule conditions but either seldom alternated their choices (Experiment 1) or greatly reduced choice alternations (Experiment 2) under the runway stay rule condition. We discussed these effects in terms of rats' natural foraging strategies and as a factor that interacts with other within- and between-series variables that affect serial pattern behavior. PMID- 15825891 TI - Cancer center benefits from collaborative approach. PMID- 15825892 TI - Flextime: hospital spaces bend to meet changing demand. PMID- 15825893 TI - Airways: assessing and upgrading OR air handling systems. PMID- 15825894 TI - Do diligence. A look at the 10 most common errors in medical equipment maintenance invoices. PMID- 15825895 TI - Suite success: applying 'universal' principles to surgical suites. PMID- 15825896 TI - Order from chaos: MSDS management systems help facilities stay on top of need-to know info. PMID- 15825897 TI - Exercise, pain, perceived family support, and quality of life in Korean patients with ankylosing spondylitis. AB - Relations of habitual exercise and pain, perceived family support, and the quality of life in patients with functional class II for ankylosing spondylitis were explored. In a cross-sectional study perceived pain, family support, and quality of life were compared for 30 patients (23 women and 7 men whose mean age was 28.3 yr.+/-8.6 yr.) practicing exercise regularly and for 38 sedentary patients (31 women and 7 men whose mean age was 27.2+/-6.7 yr.). Exercising patients reported significantly lower pain, greater perceived family support, and increased quality of life than their sedentary peers. Pain ratings were significantly negatively correlated with the quality of life in both groups (r = .26 in exercisers and r = -.50 in sedentary patients) and control group's perceived family support was significantly correlated .44 with quality of life. These results encourage further study of the associations of habitual exercise with perceived pain, family support, and quality of life. PMID- 15825898 TI - Wais-III norms for working-age adults: a benchmark for conducting vocational, career, and employment-related evaluations. AB - The Wechsler Intelligence Scales are routinely used to assess threshold variables which correlate with subsequent job performance. Intellectual testing within educational and clinical settings accommodates natural developmental changes by referencing results to restricted age-band norms. However, accuracy in vocational and career consultation, as well as equity in hiring and promotion requires the application of a single normative benchmark unbiased by chronological age. Such unitary norms for working-age adults (18- to 64-yr.-olds) were derived from the WAIS-III standardization sample in accord with the proportional representation of the seven age-bands subsumed within this age range. Tabular summaries of results are given for the conversion of raw scores to scaled scores for the working-age population which can be used to derive IQ values and Index Scores. PMID- 15825899 TI - Provision of antiretroviral prophylaxis to child rape victims in South Africa: HIV status and delayed reporting. AB - In a sample of 780 South African child victims of rape (girls under the age of 18 years; age M= 10.5 yr., SD=4.8), 10% did not qualify for antiretroviral prophylaxis because HIV status at presentation was positive and a further 34.1% did not qualify because the rape was reported more than 72 hours after the alleged offense. Regression analyses indicated that child rape victims who reported that they had been sexually active prior to the rape constitute a particularly high risk group for HIV infection and suggest primary and secondary prevention programs should be designed to decrease sexual activity among children and to encourage more immediate reporting of rape among younger children, children who are not sexually active, and children who are incestuously abused. PMID- 15825900 TI - Sex and generational differences in desired characteristics in mate selection. AB - This study compared the ranking of 18 personal characteristics or qualities which college students might use in selecting a future mate and builds on six other assessment periods from 1939-1996 which dealt with this issue. Among the most consistent findings for all seven assessment periods is the high value both sexes place on dependable character, emotional stability/maturity, pleasing disposition, and mutual attraction/love. Men across all assessment periods placed a higher value than women on good health, good cook/housekeeper, and good looks, while women placed a higher value on ambitious/industrious, similar educational background, and good financial prospect. These trends could go along with ethological theory. For both sexes chastity and similar religious background gained value; however, this may be an artifact of the current sample of 100 students. PMID- 15825901 TI - Police solidarity and tolerance for police misbehavior, revisited. AB - A small scale replication of Lester and Brink's (1985) study of police misbehavior was recently conducted with a sample of 125 police officers from Southern California. The findings of this study were diametrically opposed to those of Lester and Brink's findings. This study indicates that there does not appear to be a relationship between peer association while off duty and a proclivity for official misconduct. Several reasons are mentioned for possible explanation of the different findings. PMID- 15825902 TI - Relationship between short-term sexual strategies and sexual jealousy. AB - In a classic study, Buss, Larson, Westen, and Semmelroth reported that men were more distressed by the thought of a partner's sexual infidelity (sexual jealousy) and women were more distressed by the thought of a partner's emotional infidelity (emotional jealousy). Initially, Buss and his associates explained these results by suggesting that men are concerned about uncertainty of paternity, that is, the possibility of raising another man's child while believing the child is their own. However, later they explained the results in terms of men's preference for short-term sexual strategies. The purpose of this research was to test the explanation of short-term sexual strategies. Men and women subjects were instructed to imagine themselves in a relationship which was either short-term (primarily sexual) or long-term (involving commitment) and then respond to Buss's jealousy items. It was hypothesized that, when both men and women imagined a short-term relationship, they would be more threatened by a partner's sexual infidelity, and, when they imagined a long-term relationship, they would be more threatened by a partner's emotional infidelity. Support was found for this hypothesis. PMID- 15825903 TI - Individual characteristics associated with resilience in single-parent families. AB - This study identified individual characteristics as a resource to enhance the resilience of a family dealing with the loss of a parent. 25 white single-parent families who had lost a parent between 1 and 4 yr. previously were identified by four postgraduate students in the Western Cape, South Africa. Each single-parent, 19 women and 6 men (M age=48 yr., SD=7.65), were asked to state the personal qualities which helped the family adapt after the loss, after the Family Attachment and Changeability Index 8 and the Ego-resiliency Scale were completed. The expected positive relationship between personal resilience and family functioning could not be confirmed. However, the qualitative results indicate that optimism, perseverance, faith, expression of emotions, and self-confidence were prominent individual characteristics of resilience viewed as resources in promoting resilience in these single-parent families. PMID- 15825904 TI - Measurement error and the correlation between positive and negative affect: spearman (1904, 1907) revisited. AB - There has been considerable debate in the literature on the structure of affect, especially on the correlation between positive and negative affect and the effect of measurement error on this correlation. In this brief article, it is shown that, as Spearman (1904, 1907) noted, the extent to which the correlation between imperfect measures is attenuated by measurement error depends upon the reliabilities of the measures used. PMID- 15825905 TI - Psychometric properties of the reassurance-seeking scale in a Turkish sample. AB - This study examined the psychometric properties of the Reassurance-Seeking Scale in a sample of 102 Turkish undergraduate students. High internal consistency reliability was found for the Reassurance-Seeking Scale (alpha=.86). Factor analysis of the scale identified a single component that accounted for 71% of the total variance. The scale was significantly positively correlated with the Beck Depression Inventory and Beck Anxiety Inventory and had a significantly negative correlation with the Rosenberg Self-esteem Scale. Partial correlations of Reassurance-seeking with Depression scores as controlled by Anxiety scores and with Anxiety scores as controlled by Depression scores indicated that Reassurance seeking scores maintained association with Depression but not with Anxiety. All these findings were in line with expectations. PMID- 15825906 TI - The Spanish version of Betts' Questionnaire Upon Mental Imagery. AB - Sheehan's shortened form of Betts' Questionnaire Upon Mental Imagery is one of the most widely used inventories for measuring imaging. Participants were 562 undergraduate students (414 women and 148 men), with a mean age of 20.2 yr. In a principal components analysis followed by varimax orthogonal rotation for the Spanish version, eight factors were identified, six of which coincide with those of the original English version. Internal consistency was .92 (Cronbach alpha). Scores on the Betts Questionnaire Upon Mental Imagery correlated -.34 with those on the Gordon Test of Visual Imagery Control, .58 with those on the Vividness of Visual Imagery Questionnaire, and .02 with the Verbalizer-Visualizer Questionnaire. PMID- 15825907 TI - Effects of anxiety on communication tasks for children in traditional and individualized-multiage classrooms. AB - The effects of anxiety on concept acquisition were evaluated for first and second grade children enrolled in either a traditional or in an individualized or multiage learning program, the latter specifically designed to reduce anxiety during the learning process. Regardless of their learning environment, first graders who scored high on trait anxiety, measured by the State-Trait Anxiety Inventory for Children, performed more poorly than those with low trait anxiety. Individualized-multiage second grade children scored lower on state anxiety than children in the traditional group. The largest reduction in anxiety occurred at the second grade, suggesting that anxiety reduction was cumulative for the individualized-multiage program children during the first two years of their primary grade education. However, the individualized-multiage environment was detrimental to performance of second graders on the easier concrete concept acquisition test, suggesting that some anxiety is needed during learning to enhance attention and facilitate performance. PMID- 15825908 TI - Updating information in verbal working memory and executive functioning. AB - 50 older adults (M age=77.9 yr., SD=7.3; 35 women and 15 men) were tested using the updating working-memory task. They were also given the neuropsychological Wisconsin Card Sorting Test, assumed to evaluate executive functioning and the frontal cortex. A factor analysis with age, education, and verbal ability partialled out was computed on the updating task outcomes and resulted in a two factor solution, indicating that this task requires two independent processes, interpreted as reflecting a storage component and an updating component. Partial correlations with age, education, and verbal ability partialled out indicated that Wisconsin Card Sorting Test measures were significantly associated with the factor supposed to reflect the updating process. Such results appeared consistent with the model of working memory with a central executive system involved in the updating process and related to the executive-frontal functioning, and a phonological loop system involved in the storage of verbal information and not linked to executive-frontal functions. PMID- 15825909 TI - Type a behavior pattern as a predictor of psychiatric sick-leaves of Spanish teachers. AB - The present study examined the relationship between the Type A Behavior Pattern and certain psychopathological disorders, namely, anxiety and depression, within an occupational setting. A sample group of 100 high school teachers (37 men and 63 women) who had taken one or more psychiatric leaves of absence were studied. An additional 100 teachers (47 men and 53 women), with no history of psychiatric sick-leaves, were randomly assigned to a control group. The groups shared the same organizational setting. The Jenkins Activity Survey, Form H, was administered to assess the Type A Behavior Pattern of both groups. Binomial logistic regression analysis indicated this pattern was related to psychiatric sick-leaves. PMID- 15825910 TI - Topic variability and linguistic convergence in marital couples' speech as related to specific attachment style. AB - Links between mental working models, attachment style, and linguistic strategies were analyzed in the present research, which focused on linguistic choices made by marital couples. Eight couples (16 Italian subjects, M age=49.1, SD=0.4) with drug-addicted sons and undergoing systemic therapeutic treatment were recruited. Conversational transcripts were coded by two judges. The research analyzed (a) the definition of a topics map derived from the Adult Attachment Interview, using the discursive level of the topic; (b) the systemic-functional analysis of the topics, through both the micro- and macro-unit levels and the lexical/semantic indexes of the discourse; (c) the within-couple convergence in terms of linguistic moves, and, more generally, of their cognitive strategies. Specifically, a hierarchical model of the topic organization was used to explain the recursivity of the thematic choices made by the participants. Secondly, the characterization of the topics in terms of their cognitive/emotive values (high/low "critical" topics) was explained, especially through some "linguistic markers" as disfluency or lexical/semantic indexes (i.e., linguistic variety and completeness indexes). Third, it was found that different and specific linguistic strategies were adopted by the couples in order to reiterate their internal working models. The high convergence of linguistic patterns inside the marital couple was discussed as representative of "dysfunctional" working models, from both the communicative and relational points of view. PMID- 15825911 TI - Personal disturbance scale: factor structure confirmed in a large nonclinical sample. AB - The two-component structure of anxiety and depression items of the short form Personal Disturbance Scale, reported in an earlier clinical study of 480 adult psychiatric patients, was substantially replicated in a large nonclinical sample of 758 adults. PMID- 15825912 TI - Status of psychology as a science in northeast Brazil: undergraduate students' perceptions. AB - The Psychology as Science scale was used to assess whether Brazilian students believe psychology is a science. 190 undergraduates from four universities in Brazil participated. Analysis showed that 54% of students strongly agree that psychology is a science, 26% strongly agree that psychological research is necessary and that training in methodology is important, and 25% strongly agree that behavior is predictable. PMID- 15825913 TI - Constructive thinking inventory: evidence of validity among Iranian managers. AB - This study examined the validity and incremental validity of the Constructive Thinking Inventory in a sample of Iranian managers. These 159 men were 39.9 yr. old (SD=2.5) and volunteered to participate in a project in which they responded to the Constructive Thinking Inventory, the Big Five Factors, the Costello and Comrey Depression and Anxiety Scales, and the Perceived Stress Scale. Numerous findings confirmed the validity of the Constructive Thinking Inventory, and the Global Constructive Thinking subscale displayed incremental validity. These data supported the validity of the Constructive Thinking Inventory and its associated theoretical assumptions in a sample of Iranian managers. PMID- 15825914 TI - Foot-in-the-door and door-in-the-face: a comparative meta-analytic study. AB - The so-called foot-in-the-door and door-in-the-face techniques have been generally considered as procedures which may consistently influence others' behavior. This study compared efficiency (in terms of compliance rates with various target requests) of both techniques using meta-analysis. It is based on the accumulated studies directly comparing foot-in-the-door (n=22, subjects= 1581) and door-in-the-face (n=22, subjects= 1611) under a similar target request. Analysis indicated no significant differences in efficiency between the paradigms. PMID- 15825915 TI - Preliminary study of psychological factors affecting clinic attendance and glycemic control of Japanese patients with type 2 diabetes mellitus. AB - We studied 50 Japanese patients who were first diagnosed with type 2 diabetes mellitus. Psychological functioning was assessed by using the Rorschach test and the Yatabe-Guilford Personality test. Analysis indicated that patients who discontinued clinic attendance during the 2-yr. study period had more depression, lower general activity and less extraversion, more negative self-attitudes, and more concern about being helpless than those who continued having regular followups. This study also found that patients who had worse glycemic control 2 yr. after being first diagnosed reported more inferiority feelings, lower general activity, more need for closeness to others, and more emotional demands than those who had better glycemic control. In conclusion, understanding the psychological functioning would be helpful in the management of Japanese patients with type 2 diabetes mellitus. PMID- 15825916 TI - Women with low activity are at increased risk for depression. AB - This study assessed if there are sex differences in the relationship between amount of physical activity and depression in primary care outpatients. Participants (N= 111), referred by their family physicians, wore a heart-rate monitor for 24 hr. The two-lead monitor collected the number of beats per minute and minute-averaged horizontal movement data, e.g., from walking or moving about. All participants underwent structured clinical interviewing using the Structured Clinical Interview for DSM-IV and completed the Beck Depression Inventory-II and the Symptom Checklist-90-Revised. For the total sample, patients with low activity were more likely to be depressed (p<.006). Adults under the age of 45 (n=85) with low activity were 7.4 times more likely to be depressed than young adults who showed normal activity. However, in adults 45 or older, there was no relationship between activity and depression. In a subsample of women under the age of 55 (n=70), 91% of the low activity group and 39% of the normal activity group were diagnosed with depression (p<.003). Women in the low activity group were 15.7 times more likely to be depressed than women in the normal activity group. In contrast, there was no relationship between activity and depression in men. Additional research is needed to assess whether low activity is a potent predictor of depression in women. PMID- 15825917 TI - Frontal lobe dementia and subcortical vascular dementia: a neuropsychological comparison. AB - We compared the performance of 40 patients with frontal lobe dementia to that of 40 patients with subcortical vascular dementia (80 patients including, 46 men and 34 women) in a set of tasks assessing attentional, executive, and behavioural tasks. The frontal lobe dementia represents an important cause for degenerative disruption and is increasingly recognised as an important form (up to 25%) of degenerative dementia among individuals of late-middle-age. The main involvement is the frontal-subcortical pathway, which is the final target of impairment even in subcortical vascular dementia. A wider involvement of the cortical (decisional) layers in frontal dementia, in contrast with the prominent and widespread involvement of the subcortical pathways (refinement and corrections programs) creates the different profiles of the two groups. Frontal patients have more difficulties in abstract reasoning, focusing attention, and implementing strategies to solve problems. They exhibit more profound behavioural alterations in personality and social conduct and show only moderate depression, and a total lack of insight concerning their dinical condition. In contrast, the patients with subcortical vascular dementia have poor general cognitive functions, high insight, and important depression and apathy as the principal and most salient characteristic of their behavioral conduct. PMID- 15825918 TI - Factor structure of the self-construal scale in a Vietnamese-American adolescent sample. AB - This study investigated the factor structure of the Self-construal Scale, an important measure of interdependent and independent self-construal. An exploratory factor analysis with the principal component method and varimax rotation supported Singelis' original two-factor model (1994) for a sample of 152 Vietnamese-American high school adolescents recruited from Little Saigon in Orange County, California. Implications for the conceptualization of self construal and issues in research are discussed. PMID- 15825919 TI - Factors associated with intentions for colorectal cancer screenings in a Chinese sample. AB - This study investigated factors associated with intention to screen for colorectal cancer among Chinese people age 40 yr. and older. In-depth individual interviews were conducted. Participants were recruited from urban middle-class communities in Taichung, Taiwan through homogeneous, criterion, and snowball sampling strategies (N=32). Immersion and crystallization techniques were used in data analysis. Member check was used to ensure data quality. Analysis showed that factors related to intentions to have fecal occult blood tests were influenced by the inconvenience (61%) and the unpleasant screening procedure (29%), while participants' gastrointestinal symptoms or family history (32%) and physicians' recommendation or patients' self-health conditions (25%) were more important to intentions for a flexible sigmoidoscopy or colonoscopy screening. This study suggests tailoring strategies and messages to encourage use of various screening. Unique cultural issues, such as avoidance of facing problems (28%), embarrassment (23%), or lack of acceptance of preventive behaviors (16%), were also discussed with recommendations provided. PMID- 15825920 TI - Complementary and alternative medicine use by psychiatric inpatients. AB - 82 psychiatric inpatients hospitalized for acute care were interviewed about their use of complementary and alternative medicine (CAM) modalities. The clinical diagnoses of respondents included Depressive Disorder (61%), Substance Abuse (26%), Schizophrenia (9%), and Anxiety Disorders (5%). Analysis indicated that 63% used at least one CAM modality within the previous 12 mo. The most frequently used modality was herbal therapies (44%), followed by mind-body therapies such as relaxation or mental imagery, hypnosis, meditation, biofeedback (30%), and spiritual healing by another (30%). Physical modalities such as massage, chiropractic treatment, acupuncture, and yoga were used by 21% of respondents. CAM therapies were used for a variety of reasons ranging from treatment of anxiety and depression to weight loss. However, most respondents indicated they did not discuss such use with their psychiatrist or psychotherapist. PMID- 15825921 TI - Higher-order factors of the big five model of personality: a reanalysis of Digman (1997). AB - Based on the results from factor analyses conducted on 14 different data sets, Digman proposed a model of two higher-order factors, or metatraits, that subsumed the Big Five personality traits. In the current article, problems in Digman's analyses were explicated, and more appropriate analyses were then conducted using the same 14 correlation matrices from Digman's study. The resultant two-factor model produced improper solutions, poor model fit indices, or both, in almost all of the 14 data sets and thus raised serious doubts about the veracity of Digman's proposed model. PMID- 15825922 TI - Experience of colorectal cancer screening using a home-administered kit for fecal occult blood tests among a Chinese worksite population in Taiwan. AB - This study examined the experience of fecal occult blood test (FOBT) using a home administered kit that requires no stool handling for colorectal cancer screening among Chinese people age 40 and older. 304 participants were recruited from 10 worksites in Taiwan. Both oral and written instructions on how to use the test kit were provided. After participants completed the screening test, their experiences were documented through structured open-ended probing survey questions. Although analysis showed some challenges for Chinese participants as first time users, the overall reactions and perceived advantages towards the kit were promising. Interventions should consider the participant's stage of adaptation to new screening strategies, literacy, and environmental conditions. Since no known research has been conducted among Chinese people with a kit, experience provides valuable qualitative insights for planning intervention. PMID- 15825923 TI - Self-pluralism and dissociation. AB - The study examined the relationships among self-pluralism and dissociative experiences. 100 students were assessed using the Self-pluralism Scale and the Dissociative Experiences Scale. Scores on self-pluralism were positively correlated with others: overall Dissociation (r=.50), Absorption (r=.54), Depersonalization (r=.43) and Amnesia (r = .28). PMID- 15825924 TI - Predicting attitudes toward operation Iraqi freedom. AB - 98 male and female college students ranging in age from 17 to 22 years (M= 19.2, SD= 1.1) were administered a questionnaire that contained Altemeyer's 20-item Right-Wing Authoritarianism Scale (RWA), questions about the legitimacy of the 2000 presidential election, Mayton's Physical Nonviolence Subscale, and a scale designed to measure attitudes toward Operation Iraqi Freedom. RWA scores, perceptions of the legitimacy of the 2000 presidential election, and scores on the Physical Nonviolence subscale were independent predictors of attitudes toward Operation Iraqi Freedom. In addition, those who had higher RWA scores were more likely to perceive the 2000 presidential election as legitimate. There was no significant correlation between RWA scores and attitudes toward physical nonviolence. PMID- 15825925 TI - Intensity and prevalence of current fears among Iranian university students. AB - The intensity and prevalence of fears were assessed in Iranian university students by applying the Fear Survey Schedule for Children, II. Analysis indicated that the intensity and prevalence of fears varied by students' sex, field of study, and birthplace. Results are discussed in relation to previous findings, and a few recommendations together with some educational implications were noted. PMID- 15825926 TI - Neuropsychological correlates of Greek alcoholic patients who report memory disturbances. AB - We studied 40 male and 37 female (M age=63 yr.) Greek alcoholic patients and an equal number of control subjects. Both groups were evaluated with the Mini-Mental State Examination, the Syndrome Short Test, the Verbal Fluency Test (Category & Letter), the Clock Test, and the Digit Span (Forward and Backward from the Wechsler Adult Intelligence Scale-Revised). Alcoholic patients had statistically significant lower scores on MMSE, Verbal Fluency Test, and Digit Span, and higher scores on the Syndrome Short Test, while positive correlations were found among MMSE, Verbal Fluency Test, Clock Test, Digit Span-Backward, and age. These findings point to frontal lobe dysfunction in Greek alcoholic patients which is not different from that shown in patients from diverse ethnic and cultural backgrounds. PMID- 15825927 TI - Knowledge and fear of developing Alzheimer's disease in a sample of healthy adults. AB - Alzheimer's disease is the fourth leading cause of death among seniors but its insidious onset makes it difficult to distinguish from age-related decline and little is known about how much knowledge and fear people entertain about this disease. This study examined knowledge and fear of Alzheimer's disease by surveying 127 young (M= 19.6 yr.) and 118 older (M=67.9 yr.) participants. Younger participants obtained a score of 64%, while older participants obtained 58% on the knowledge test. Younger participants reported significantly higher fear overall. Knowledge and Fear scores were not significantly correlated with having a family member or knowing someone with Alzheimer's disease. PMID- 15825928 TI - Differences in readers' response towards advertising versus publicity. AB - Publicity is assumed to have higher credibility than advertising prompting communication specialists to call for its use alongside traditional advertising to achieve overall communication goals. To test the differences in readers' response towards advertisements and publicity, a sample of students (N= 104; 52 men and 52 women) were shown printed messages identified as advertising or publicity. The publicity format appeared to be associated with deeper processing because participants had higher recall, more message-relevant thoughts, and better discrimination for the publicity message. In addition, participants rated the publicity message higher on Source Credibility, Brand Attitude, and Purchase Intent than a comparable advertisement. PMID- 15825929 TI - Preliminary study of police experience of misconduct in their colleagues. AB - A pilot study of 33 male and 2 female municipal police officers, most between 30 and 40 years old, found that those whose colleagues more often engage in misconduct experience more excitement and more stress in their work. PMID- 15825930 TI - The relationship of character structure to persuasive communication in advertising. AB - The persuasive effect of character structure--defined as a person's organized set of drives, dispositions, and satisfactions with which they approach the world- was assessed in the context of printed advertising. Subjects were exposed to one of two levels of argument strength (strong versus weak) and one of two levels of message spokesperson (celebrity versus noncelebrity) in a printed-advertising task. Subjects classified as Other-directed, individuals who possess a strong need to get along with others, exhibited greater attitudinal responsiveness to the test advertisement as measured on a composite attitude scale than did those classified as Inner-directed, needing to get ahead, or succeed. Other-directed subjects' attitude scores were more responsive to the message source than were the comparable scores of Inner-directed subjects. Results regarding argument strength were not significant but suggest that Other-directed attitudes are influenced by a combination of textual arguments and social cues. PMID- 15825931 TI - Cognitive self-consciousness and meta-worry and their relations to symptoms of worry and obsessional thoughts. AB - The relation between cognitive self-consciousness and meta-worry, and their association with symptoms of worry and obsessional thoughts were examined. 53 undergraduate students completed the expanded version of the Cognitive Self consciousness Scale, the Meta-worry subscale of the Anxious Thoughts Inventory, the Penn State Worry Questionnaire, and the Padua Inventory-Revised. Analysis showed that cognitive self-consciousness and meta-worry were moderately correlated (r =.57). Further, both constructs were positively associated with symptoms of worry and obsessional thoughts. When controlling for cognitive self consciousness, metaworry remained significantly correlated to both types of symptoms. Yet, when controlling for meta-worry, correlations between cognitive self-consciousness and symptoms of worry and obsessional thoughts clearly attuned and were no longer significant. These findings suggest that meta-worry is more important for understanding excessive, intrusive thought patterns than the mere tendency to monitor one's thoughts. PMID- 15825932 TI - Students' ratings of teaching effectiveness: a laughing matter? AB - Gump in 2004 identified a positive significant relationship between awareness of daily class objectives and ratings of the instructor's overall teaching effectiveness. The idea that rating of teaching effectiveness can be related to other non-teaching related attributes of the lecturer was further examined. Correlations based on ratings of teaching effectiveness from 453 undergraduate students (M=21 yr., SD= 5.5; 73% women) showed that another nonteaching related variable, namely, how funny the instructor was perceived, was significantly related to indicators of teaching effectiveness. PMID- 15825933 TI - Child molestations by homosexual foster parents: Illinois, 1997--2002. AB - Do those who engage in homosexuality disproportionately sexually abuse foster or adoptive children as reported by child protective services? Illinois child services reported sexual abuse for 1997 through 2002. 270 parents committed "substantiated" sexual offenses against foster or subsidized adoptive children: 67 (69%) of 97 of these mother and 148 (86%) of 173 of these father perpetrators sexually abused girls; 30 (31%) of the mothers and 25 (14%) of the father perpetrators sexually abused boys, i.e., 92 (34%) of the perpetrators homosexually abused their charges. Of these parents 15 both physically and sexually abused charges: daughters by 8 of the mothers and 4 of the fathers, sons by 3 of the mothers, i.e., same-sex perpetrators were involved in 53%. Thus, homosexual practitioners were proportionately more apt to abuse foster or adoptive children sexually. PMID- 15825934 TI - Young children's natural learning environments: contrasting approaches to early childhood intervention indicate differential learning opportunities. AB - 1,000 parents of infants and toddlers enrolled in early childhood intervention programs were surveyed about the number of learning opportunities provided their preschool children using different approaches to early intervention. Findings showed that more learning opportunities were reported when participation in everyday activity settings was conceptualized as a type of intervention rather than as settings for professionals to conduct their interventions. PMID- 15825935 TI - Relationship between Machiavellianism scores and performance of real estate salespersons. AB - Data from two samples (ns=37 and 35) of real estate agents showed a significant positive correlation of .37 between Machiavellianism (Mach-B scores) and self reported sales volume. Present findings support earlier results from samples of stockbrokers and automobile salespersons showing Mach-B scores to be positively related to sales performance. PMID- 15825936 TI - University faculty preparation of students in using natural environment practices with young children. AB - 155 university faculty teaching students in physical therapy, occupational therapy, speech and language pathology, early childhood special education, or multidisciplinary studies programs were surveyed to assess how the students were taught how to use everyday family and community activities as natural learning opportunities for young children. Analysis showed that the faculty provided very little training in using community activity settings as contexts for children's learning and that physical therapy faculty provided less training in using natural environments as sources of children's learning opportunities than faculty in the other disciplines. PMID- 15825937 TI - Gang violence: too high a price to pay. Integris Health, Oklahoma City. PMID- 15825938 TI - Domestic violence: the silent killer. Fairview Hospital and Health System, Minneapolis. PMID- 15825939 TI - Hospital governing boards and quality of care: a call to responsibility. PMID- 15825940 TI - One giant leap for quality. When boards get behind quality initiatives, patient care benefits. AB - When boards get behind quality initiatives, patient care definately benefits--but which measures help most? Following is an analysis of how well the Leapfrog Group's guidelines are working at participating hospitals. PMID- 15825941 TI - Physicians in governance: their unique learning curve. PMID- 15825943 TI - The gift of life. PMID- 15825942 TI - Effective hospital governance: compliance risk areas. PMID- 15825944 TI - Hospitals compare quality. CMS' release of quality data may change behavior. PMID- 15825945 TI - Reversal of fortune. After years of scaling back beds because of low occupancy rates, behavioral health facilities are beginning to make a comeback. AB - For decades states have been cutting behavioral health beds because of low occupancy. But a recent survey showed occupancy rates at these hospitals at record highs, and analysts expect more consolidation as larger providers expand. A change in federal payment methods favors free-standing facilities. "The free standing units will do better as a group," says the National Association of Psychiatric Health Systems' Mark Covall. PMID- 15825946 TI - Pushing procedures. Study condemns ads by academic medical centers. PMID- 15825947 TI - Facing the music. Multihospital systems adopting Sarbanes-Oxley changes. PMID- 15825948 TI - Poor performance. Insurance group's ratings system angers providers. PMID- 15825949 TI - Docs, seniors get more bad news. Medicare premiums to rise even more than expected. PMID- 15825950 TI - Broke and banished. Pending bill would make exclusion by CMS easier. PMID- 15825951 TI - For-profits for sale. Investor-owned hospital chains choosing to sell, not buy. PMID- 15825952 TI - On the job. Nurses reveal most, least enjoyable aspects of work. PMID- 15825953 TI - Record settlement. PharMerica resolves kickback allegations. PMID- 15825954 TI - Back in action. Feds force hospitals to end market agreements. PMID- 15825955 TI - Risky business. Okla. hospitals may launch liability insurance company. PMID- 15825956 TI - Va. hospital exits Carilion. PMID- 15825957 TI - A long, sad week. Healthcare organizations fail to voice opinions in Schiavo debate. PMID- 15825958 TI - A study in what not to do. Schiavo case reveals dangers of letting strangers make end-of-life decisions. PMID- 15825959 TI - Armed with innovation. Israel has become a prolific developer of new medical technologies and the nation's dominant defense industry is often the incubator. PMID- 15825960 TI - By the numbers. Top nursing facility chains ranked by number of beds as of Dec. 31, 2003. PMID- 15825961 TI - Evolving demographics and disparities in an urban diabetes clinic: implications for diabetes education and treatment. AB - OBJECTIVES: To compare demographics and disease characteristics in a multiethnic diabetes clinic population to identify changes over time. DESIGN: Analysis and comparison of demographics and disease characteristics of diabetes patients, recorded electronically at intake over 10 years. SETTING: An urban outpatient diabetes clinic. PATIENTS: A total of 8,551 African-American (88%), White (7%), or Hispanic (3%) patients (average age, 52 years; mean diabetes duration, 5.1 years; 59% women). MAIN OUTCOME MEASURES: Proportion of patients by ethnic group, age, diabetes duration, initial hemoglobin A1c, and body mass index. RESULTS: Between 1992 and 2001, the percentage of African-American patients was relatively unchanged (from 87.6% to 87.2%; P=.2), White patients decreased (from 9% to 5%; P=.0006), and Hispanic patients increased (from 1.3% to 5.5%; P<.0001). Among African-American patients, average age decreased from 52 to 50 years (P=.015), diabetes duration decreased from 5.6 years to 4.3 years (P=.0003), initial hemoglobin A1c decreased from 9.3% to 8.8% (P<.0001), and body mass index increased from 31 kg/m2 to 32.1 kg/m2 (P=.0001). Compared with African-American and White patients, Hispanic patients were younger (P<.0001) and had a lower body mass index (P<.0001) but had hemoglobin A1c comparable to that of African American patients (9.3% vs 9.1%; P=.45) and higher than that of White patients (9.3% vs 8.7%; P=.0022). CONCLUSIONS: The demographic and disease profiles of patients in this urban diabetes clinic have shifted, and disparities in glycemic control and obesity exist. Modifications in treatment and education approaches may be necessary to compensate for a changing patient population. PMID- 15825962 TI - Chronic pain in older Black Americans: the influence of health and psychosocial factors. AB - Chronic pain may differentially affect racial and ethnic minorities while diminishing their mental well-being and physical health. Although the literature documents the impact of chronic pain on physical, psychological, and social well being in White Americans, the impact of chronic pain in Black Americans has not been extensively investigated. This study examined the relationship between pain intensity, health variables, various demographic characteristics, and psychosocial factors in a sample of older Black Americans (N=189). Participants were older, community-dwelling Black Americans with a mean age of 69.8+/-9.12 years. Results showed that reporting more depressive symptoms, greater physical impairment, and being younger were significant indicators of pain intensity and accounted for 27% (R=.52) of its variation. These results suggest that pain adversely affects the psychological and social well-being in some older Black Americans. This investigation emphasizes the need for further studies examining within-group differences of the pain experience. This research may help develop models that assess how social, cultural, environmental, physical and psychological health factors influence the daily experience of pain in racially and ethnically diverse populations. PMID- 15825963 TI - Addressing oral health disparities in settings without a research-intensive dental school: collaborative strategies. AB - Research suggests that oral health is linked to systemic health, and those with poor oral health are potentially at greater risk for important diseases, including cardiovascular disease, stroke, diabetes mellitus, and adverse pregnancy outcomes. Asians and Pacific Islanders (APIs) in Hawaii have high rates of many such diseases. Studies in children in Hawaii have revealed disparities in dental health; for example, API children have significantly higher rates of cavities than other groups. Hence, conducting further study is vital in adults, particularly APIs, to assess oral health and its correlation to overall health outcomes. Given the lack of a dental school and the lack of fluoridated water in the state, the University of Hawaii's John A. Burns School of Medicine (ABSOM) has identified the need to assume a leadership role in creating effective community-based oral health research and treatment programs. With the support of the National Institute of Dental and Craniofacial Research, JABSOM fostered a collaborative relationship with the University of North Carolina at Chapel Hill School of Dentistry, a premiere research-intensive dental school, the Waimanalo Health Center, and the Hawaii State Department of Health. This partnership has worked together to implement a community-based approach to performing research designed to illuminate disparities and develop innovative strategies to promote oral health in Hawaii's diverse populations. We hope that this collaborative, culturally competent approach may serve as a model for use in other settings without a research-intensive dental school. PMID- 15825964 TI - Endothelial nitric oxide synthase intron 4 polymorphism is a marker for coronary artery disease in African-American and Caucasian men. AB - OBJECTIVES: We investigated the association of the intron 4 polymorphism of the endothelial nitric oxide synthase (eNOS) gene with coronary artery disease (CAD). BACKGROUND: Genetic alterations in the gene encoding for eNOS could contribute to the development and progression of CAD. METHODS: We genotyped for the eNOS intron 4 polymorphism in 194 subjects undergoing coronary angiography. Genotyping was performed with polymerase chain reaction-restriction fragment length polymorphism for the variable number of tandem repeats in intron 4. Coronary artery disease (CAD) was assessed by quantitative coronary angiography, and endothelial function was measured by brachial ultrasonography. We performed logistic regression analysis for the effect of eNOS intron 4 polymorphism and other coronary risk factors on multi-vessel CAD and endothelial function. RESULTS: The 4a-allele frequency in African Americans was 0.31, while the 4a-allele frequency in Caucasians was 0.15 (P<.001). The prevalence of the 4a-allele was highest among subjects with multi-vessel disease both for African Americans and for Caucasians. A race-adjusted comparison of the prevalence of the 4a-allele among subjects with multi-vessel disease vs those without was statistically significant (P=.03). No correlation was found between the eNOS intron 4 polymorphism and endothelial function. CONCLUSIONS: The eNOS intron 4 polymorphism may be a marker of multi vessel CAD in African Americans and Caucasians. PMID- 15825965 TI - Race-ethnic differences in the extent, prevalence, and progression of coronary calcium. AB - OBJECTIVE: To compare across four race-ethnic groups the baseline prevalence and extent of coronary calcium and the 7-year rate of progression in the extent of coronary calcium. DESIGN: The South Bay Heart Watch is a prospective cohort study designed to appraise the value of coronary calcium for predicting cardiovascular outcomes in asymptomatic adults with cardiac risk factors. Statistical analyses were conducted to evaluate ethnic differences in the prevalence, extent, and progression of coronary calcium among Caucasian, African-American, Hispanic, and Asian participants. SETTING: Population-based study. PATIENTS OR PARTICIPANTS: Between December 1990 and December 1992, 1289 participants without coronary heart disease underwent baseline risk factor screening and computed tomography for coronary calcification (Cohort 1). Seven years later, 828 (64%) participants returned for follow-up evaluation and re-scanning (Cohort 2). MAIN OUTCOME MEASURES: Prevalence, extent, and progression of coronary artery calcium. RESULTS: In Cohort 2, compared to Whites, African Americans had a lower prevalence of coronary calcium at baseline (P=.012) and follow-up (P=.005), smaller calcium scores at baseline (P=.005) and follow-up (P=.0004), and less progression (P=.001); Hispanics had a lower prevalence of coronary calcium at follow-up (P=.04) with smaller calcium scores (P=.011), and less progression (P=.009). In contrast, no differences were seen between Whites and Asian/Pacific Islanders. Race-ethnic differences in progression persisted after adjusting for risk factors and participation bias (P<.05). CONCLUSIONS: The present results lend further credence to the notion that race-ethnic differences exist in the prevalence and rate of progression of coronary calcification. The relationship between calcification and the incidence of coronary heart disease in these race ethnic groups needs further exploration. PMID- 15825966 TI - Racial and ethnic disparities in the pharmacologic management of diabetes mellitus among long-term care facility residents. AB - OBJECTIVE: To evaluate the prevalence of racial and ethnic disparities of antidiabetic treatment among residents of long-term care facilities in five states. RESEARCH DESIGN AND METHODS: Retrospective, cross-sectional study of 50,427 elderly nursing home residents with diabetes in New York, South Dakota, Kansas, Mississippi, and Ohio between 1993 and 1997. RESULTS: Thirty to fifty percent of residents received no antidiabetic medications. After adjusting for sociodemographic characteristics, comorbid conditions and diabetes severity, Blacks and Hispanics had lower rates of antidiabetic medication use than Whites, while Asians had slightly higher rates. For Native Americans the results were mixed, little disparity was seen when compared with Whites observed among New York nursing home residents, and while in South Dakota Native Americans had significantly lower rates of antidiabetic medication use than Whites. CONCLUSION: Although lack of information on glycemic control and non-medical treatments do not allow us to comment on quality of diabetes care, more research is needed to understand why some nursing homes residents are less likely to receive antidiabetic medication. PMID- 15825967 TI - Risk factors for coronary heart disease among immigrant women from Iran and Turkey, compared to women of Swedish ethnicity. AB - OBJECTIVES: The aim of the study was to compare a group of immigrant women from the Middle East living in Sweden to Swedish-born controls regarding the prevalence of certain cardiovascular risk factors. DESIGN: Health survey of randomly selected foreign-born women and women native in Sweden. SETTING: Uppsala, Sweden. SUBJECTS: A total of 107 immigrant women aged 35-64 years from the Middle East (Iran [N=71] and Turkey [N=36]) living in Uppsala and residents in Sweden for at least three years and a control group of ethnic Swedish women (N=50). MAIN OUTCOME MEASURES: A questionnaire and a clinical examination specially directed towards measuring cardiovascular risk factors and prevalence of obesity, diabetes mellitus, and hypertension. RESULTS: A less beneficial cardiovascular risk profile was found among immigrant women than among ethnic Swedish women. Turkish women had substantially higher body mass index (BMI), larger waist, higher waist/hip ratio and sagittal abdominal diameter, higher levels of serum triglycerides, and lower HDL cholesterol concentration compared with Swedish-born women. A similar tendency was seen also for Iranian women. CONCLUSION: The present study shows important ethnic differences in cardiovascular disease risk factor pattern. Immigrant women from Iran and Turkey are heavier than women born in Sweden and have a higher prevalence of abdominal obesity, an unfavorable lipid profile, and a high degree of physical inactivity during leisure time, which may predispose for a higher incidence of diabetes and atherosclerotic cardiovascular disease. PMID- 15825968 TI - Genetic predictors of coronary heart disease risk factors in premenopausal African-American women. AB - PURPOSE: This study examined polymorphisms at the loci HLA-DQB1, HLA-DRB1, TNFa microsatellite, and D6S89, all which lie within or telomeric to the major histocompatibility complex (MHC) and the apolipoprotein E (APOE) and THO-1 loci in premenopausal African-American women (AAW) for their association with known coronary heart disease (CHD) risk factors. The sample, drawn from community and military sources, included premenopausal AAW (mean age=34.18) who were at low risk (n=117) and high risk (n=173) for CHD. METHODS: In this case- (high risk) control (low risk) study, venous blood was used for DNA extraction. Polymorphisms were assessed by using a variety of standard polymerase chain reaction (PCR) methods. Allelic controls were used in all reactions, and two individuals sized and concurred on allele assignment in each analysis. Vertical auto profile (VAPI), glucose challenge tests, measurement of insulin levels, blood pressure, body mass index (BMI), and waist-hip ratio (WHR) assessments were conducted by using standardized procedures. Pearson's correlation coefficients and assessment of allele distributions via relative frequency and frequency variance were conducted in relation to military status, risk group, and risk factors by using exact P values and likelihood ratio chi-squared (Irchi2) statistic. The significance level was set at .05. RESULTS: Of the 237 women (low and high risk), 116 of the women in the sample were in Stage I obesity or heavier. Of 237 women (low and high risk), 85 (36%) of the women in this sample were insulin resistant. The frequency of D6589 allele 185, D6S89 allele 191, TNFa allele 97, and TNFa allele 103 alleles were higher in the high-risk than the low-risk group; and the D6S89 195 allele was higher in the low-risk group. Elevated systolic blood pressure (SBP) was associated with HLA-DRB1*09 and TNFa 117 alleles. APOE*4, TNFa 109, and DRB1*107 alleles were associated with increased relative risk for elevated total cholesterol to high-density lipoprotein (HDL) ratios. APOE*4 and D6589 193 alleles were associated with an elevated risk for low-density lipoprotein (LDL) or LDL sub-fraction levels. APOE*2 was associated with a lower relative risk for total cholesterol to HDL ratios. Metabolic syndrome was identified in 26.6% of the sample and was associated with the presence of DRB1*09, DRB1*12, and DRB1*15 alleles. Lp(a) levels were positively associated with risks for HDL, HDL2, HDL3, LDL, and total cholesterol. Lp(a) was negatively associated with risks for very low-density lipoprotein (VLDL), triglyceride, fasting blood sugar (FBS), insulin resistance, SBP, weight, and WHR. CONCLUSION: The association of APOE, DRB1, D6589 and TNFa alleles with risk of CHD suggest that these are candidate genes or linked to genes for CHD in this cohort of AAW. Our data supported elevated plasma Lp(a) as a potential risk factor in AAW; however, its role is still unclear. The premenopausal AAW in this sample had a higher than expected rate of metabolic syndrome, which was associated with DRB1 alleles. PMID- 15825969 TI - Metabolic syndrome in a multiethnic population in rural Hawaii. AB - OBJECTIVE: The National Cholesterol Education Program Adult Treatment Panel III diagnostic criteria for metabolic syndrome (MS) provide a standard for comparing various populations. Using these criteria, the Third National Health and Nutrition Examination Survey reported an overall US prevalence of 21.8%. With these same criteria, we estimated the prevalence of MS among a multiethnic population in rural Hawaii. DESIGN: These data are from a cross-sectional survey from 1997-2000. SETTING: The survey was conducted in the rural community of North Kohala. PARTICIPANTS: More than 1,450 adult residents from five ethnic categories were included: Caucasian, Japanese, Filipino, Hawaiian/part-Hawaiian, Other/mixed non-Hawaiian. Ethnic ancestry was determined by self-report. Ethnic differences were compared by using logistic regression. MAIN OUTCOMES: Blood pressure, height, weight, and waist circumference, fasting and two-hour post-oral glucose challenge plasma was obtained for lipid and glucose determinations. RESULTS: Overall prevalence was 33.4%. Prevalence was significantly higher among all ethnic groups when compared to Caucasians. Despite significant differences in the prevalence of overweight and abdominal obesity, the prevalence of MS was similar in all non-Caucasian ethnic groups. Filipinos had the highest adjusted odds for prevalent MS (prevalence OR=4.2; 95% CI=2.4-7.3). CONCLUSION: Metabolic syndrome (MS) prevalence was high in Asian ethnic groups previously reported to have low cardiovascular disease (CVD) mortality. These findings suggest either a differential effect of CVD risk factors on mortality among some ethnic groups, or more likely, that future mortality rates will increase among those ethnic groups that currently enjoy low mortality rates. PMID- 15825970 TI - Dietary intake, perceptions regarding body weight, and attitudes toward weight control of normal weight, overweight, and obese Black females in a rural village in South Africa. AB - OBJECTIVES: To determine dietary intake and attitude toward weight control of normal weight, overweight, and obese Black women. DESIGN: Cross-sectional survey. SETTING: Rural village in KwaZulu province, South Africa. PARTICIPANTS: A convenience sample (N=187) of 25- to 55-year-old women, of whom 28.9% (N=54) were of normal weight, 41.2% (N=77) were overweight, and 29.9% (N=56) were obese. MAIN OUTCOME MEASURES: Dietary intake, attitude toward weight control, perceived causes of overweight, perceived health risks, and self-reported health status. RESULTS: Dietary intake did not differ among the three groups. Overweight and obese women did not view weight control differently than women of normal weight. Most women were unconcerned about their weight. Only 2% of overweight and 30% of obese women thought they were too fat. Most women (96%) agreed that obesity was caused by biological disorders, while 39% and 9% agreed that it was caused by poor eating habits and eating too much food, respectively. Most women did not recognize the relationship between food consumed and degenerative diseases. Compared to normal-weight and overweight women, obese women had a significantly higher prevalence of self-reported high blood pressure and suffered more from back pain. CONCLUSIONS: Most of these rural women were unconcerned about their weight and most overweight and obese women did not want to lose weight. The acceptance of overweight and obesity can hinder the effectiveness of weight control programs. A special attempt should be made to target such programs to rural woman. PMID- 15825971 TI - Church-based obesity treatment for African-American women improves adherence. AB - This study compared adherence to Behavioral Choice Treatment (BCT), a 12-week obesity treatment program that promotes weight loss and exercise, among 22 Caucasian-American and 10 African-American overweight women in a university setting to 10 African-American overweight women in a church setting. Behavioral Choice Treatment (BCT) promotes moderate behavior change that can be comfortably and therefore permanently maintained. Participants obtained feedback from computerized eating diaries and kept exercise logs. Results indicated that both university groups exhibited comparable eating pathology at pre- and post treatment and comparable weight loss, despite the African-American sample attending fewer sessions. The African-American church group exhibited less disordered eating attitudes, less interpersonal distrust (eg, reluctance to form close relationships or sense of alienation) at pre-treatment, and experienced significantly greater weight loss than either university group. All groups lost weight and maintained these losses at 12-month follow-up. Preliminary results suggest treatment setting may play an important role in treatment adherence and sample characteristics. PMID- 15825972 TI - Physical activity of Aboriginals with type 2 diabetes: an exploratory study. AB - Given the magnitude of the diabetes epidemic among Canadian Aboriginals and the corresponding need to develop physical activity interventions, the aims of this study were to: 1) examine the meaning of physical activity; 2) assess physical activity behavior levels; and 3) examine the association of key Social Cognitive Theory (SCT) constructs with physical activity behavior. Thirty-four Aboriginals with diabetes completed a survey composed of questions regarding: 1) the perceived meaning of physical activity; 2) physical activity behavior; and 3) SCT constructs. An emerging theme revealed that some participants perceived physical activity leisure-time activities as appropriate across the lifespan, while the majority perceived leisure-time activities to be only for youth. Based on the reported energy expenditure estimates, 61.5% of participants were categorized as sedentary. However, when occupational and household activities were taken into account, 33.0% were categorized as sedentary. Bivariate correlations revealed that no SCT constructs were significantly associated with energy expenditure scores. Results suggest that specific SCT construct items may help understand physical activity behavior change. PMID- 15825973 TI - Alcohol use and adherence to prescribed therapy among under-served Latino and African-American patients using emergency department services. AB - Non-adherence to prescribed therapy is a factor that results in many patients attending emergency departments. However, increasingly, we recognize that patients who seek health care from inner-city emergency departments are among those experiencing health disparities. For these marginalized patients, emergency departments can play an important role in complementing the process of individualized care to achieve successful health outcomes. Research that examines socio-psychological characteristics and correlates of healthcare utilization of emergency department service users among under-served minority patients is needed for redirecting appropriate care among this segment of our population. OBJECTIVE: This study examines the prevalence and correlates of adherence to prescribed therapy among patients presenting to an inner-city emergency department for a medical emergency or trauma injury. Specific attention is paid to the role of alcohol. METHOD: A cross-sectional face-to-face survey was conducted among a consecutive sample of 412 Hispanic and African Americans aged 18 years and older who sought care at an inner-city emergency department facility. RESULTS: Among patients who use medication (N=145), 32% reported low adherence. More than 24% scored positive for alcohol problems based on the Alcohol Use Disorders Identification Test (AUDIT). Results of multivariate regression analysis showed that those emergency department patients who: 1) identified themselves as Hispanic, 2) reported excessive use of alcohol, 3) reported a higher number of depressive symptoms, and 4) reported a higher number of alcohol related consequences were less likely to adhere to prescribed therapy. CONCLUSION: Future investigation regarding factors related to adherence to prescribed medication among emergency department patients should take into consideration the role of alcohol and depression in this process. Further research is also needed to factor in patient's ethnicity in the context of treatment adherence. PMID- 15825974 TI - Influence of nativity on neonatal survival of Black twins in the United States. AB - OBJECTIVE: To determine the association between maternal nativity and neonatal survival of twins among Black mothers. METHODS: We conducted a retrospective cohort study of twin live births to Black mothers in the United States from 1995 through 1998. We compared levels of overall, early, and late neonatal mortality between twins of US-born and those of foreign-born Black mothers by using hazard ratios generated from a Cox Proportional Hazards Regression model. We adjusted for dependence of observations within twin clusters by means of the Robust Sandwich Estimator. RESULTS: A total of 70,884 individual twin live births to US born (64,035) and foreign-born (6,849) mothers were analyzed. Twins of US-born mothers had a 23% higher likelihood of dying within the neonatal period compared to those of foreign-born mothers (hazard ratio [HR]=1.23; 95% confidence interval [CI]=1.04-1.46). The disparity in neonatal demise occurred exclusively in the early neonatal period (HR=1.29; 95% CI, 1.06-1.50), with mortality indices comparable in the late neonatal period (HR=0.96; 95% CI, 0.68-1.35). Low and very low birth weight (P<.0001), preterm and very preterm (P<.0001), and small-for gestational-age neonates (P<.0001) were more prevalent among twins of US-born mothers. CONCLUSIONS: Compared to those of foreign-born, twins of US-born Black mothers experienced higher mortality in the neonatal period. The mortality disadvantage resulted mainly from lower gestational age at birth and the preponderance of small-for-gestational-age babies among US-born Black mothers. PMID- 15825975 TI - Using focus groups of older African Americans and Latinos with diabetes to modify a self-care empowerment intervention. AB - OBJECTIVE: To make cultural and age-specific modifications to a self-care empowerment intervention. DESIGN: Focus groups with self-administered surveys. SETTING: Two public health diabetes and geriatrics clinic and two senior centers in and around South Los Angeles. PARTICIPANTS: African Americans and Latinos aged > or = 55 years with diabetes (N=79), and two groups of health educators (N=16). MAIN OUTCOME MEASURES: After describing the proposed intervention, the focus group facilitators asked participants: 1) whether the community of interest would be interested in the proposed empowerment intervention; 2) why or why not; and 3) how the intervention could be modified to better address the concerns of the study community of interest. All groups were audiotaped, transcribed, and (when appropriate) translated into English. Three independent investigators read all transcripts and completed standardized coding forms for each transcript. In addition, at the end of each focus group, all participants completed a self administered written survey asking them to rate aspects of the proposed intervention. RESULTS: Older African Americans and Latinos endorsed the intervention but desired an expanded dietary educational component and identified disability as an important missing content area. Participants rejected the use of an audio learning tool and did not believe that matching group-facilitator sociodemographic characteristics was important as long as facilitators demonstrated cultural competency. CONCLUSIONS: These findings illustrate a model of participatory research in which researchers and community members work together to develop an empowerment intervention that will meet community needs and will have greater cultural appropriateness. Modifying the intervention in accordance with these findings should enhance the relevance and impact of the self-care intervention. PMID- 15825976 TI - Perceptions of African-American culture and implications for clinical trial design. AB - OBJECTIVE: To identify unique cultural variables for African Americans that might limit the effectiveness of behavioral interventions in clinical trials. DESIGN: Focus group discussions lasting 90 minutes. SETTING: Outpatient, clinical research center. PATIENTS: Twenty-six African-American men and women, who completed the screening process but were ineligible for the PREMIER study, participated in six focus group sessions. PREMIER is a multicenter, randomized clinical trial that studies the effects of three different lifestyle interventions designed to reduce blood pressure without medication. MEASUREMENTS AND MAIN RESULTS: Participants used a value sort of cultural characteristics to select items that make them unique as African Americans. The following seven themes were consistently identified: 1) extensive use of nontraditional support systems; 2) general mistrust of European Americans; 3) African Americans' being undervalued as human beings and members of American society; 4) effective use of improvisation; 5) uneven playing field as a result of persistent discrimination; 6) preservation of a unique ethnic identity; 7) socioeconomic status as a major influence and predictor of behaviors. CONCLUSIONS: Cultural variables can affect African-American perceptions of the feasibility of certain behavior modifications as health interventions and their perceptions of clinical research. Using these themes, investigators can design trials and interventions that capitalize on certain cultural variables and avoid strategies that conflict with others. The identification of such cultural characteristics unique to African Americans may help to enhance the outcomes achieved by African Americans in clinical trials, improving the generalizability of results from behavior modification research. PMID- 15825977 TI - The informed consent process in a cross-cultural setting: is the process achieving the intended result? AB - This report is based on the experiences of Navajo interpreters working in a diabetes clinical trial and describes the problems encountered in translating the standard research consent across cultural and linguistic barriers. The interpreters and a Navajo language consultant developed a translation of the standard consent form, maintaining the sequence of information and exactly translating English words and phrases. After four months of using the translated consent, the interpreters met with the language expert and a diabetes expert to review their experiences in presenting the translation in the initial phases of recruitment. Their experiences suggest that the consent process often leads to embarrassment, confusion, and misperceptions that promoted mistrust. The formal processes that have been mandated to protect human subjects may create barriers to research in cross-cultural settings and may discourage participation unless sufficient attention is given to ensuring that both translations and cross cultural communications are effective. PMID- 15825978 TI - Prevalence and predictors of smoking behavior among Samoans in three geographical regions. AB - OBJECTIVES: Provide comprehensive data on smoking behavior among Samoans. DESIGN: Cross-sectional, using systematic random sampling procedures, and in-person interviews. SETTING: US Territory of American Samoa, Hawaii, and Los Angeles, California. PARTICIPANTS: 1834 adult, non-institutionalized Samoans. INTERVENTION: None. MAIN OUTCOME MEASURES: Prevalence and independent predictors of smoking. RESULTS: Approximately one fourth (26.6%) of Samoans were current smokers, with 31.4% of men and 22.5% of women currently smoking. More current smokers were found in American Samoa (28.9%), followed by Hawaii (26.9%) and Los Angeles (24.1%, P<.001). At each site, Samoan men compared with the women were significantly more likely to smoke (P<.001), initiate smoking earlier (P<.05), and smoke more cigarettes (P<.05). Cessation rates for the sample were very low. Predictors of smoking included being younger, male, married, less educated, with lower income, and more acculturated. CONCLUSION: The high smoking and low cessation rates indicate that smoking-related diseases will be significant causes of morbidity and mortality for Samoans for many years. The findings further underscore the importance of documenting smoking patterns and their determinants for subgroups rather than aggregates such as Asian American and Pacific Islanders. If Samoans are to meet the Healthy People 2010 tobacco goals, there is a need to: 1) develop tailored tobacco awareness and cessation programs based on the recommendations made by the Taskforce on Community Preventive Services; 2) understand the complex interactions between social, cultural, and psychological determinants of smoking and cessation behaviors; and 3) develop policies to limit availability of tobacco, environmental exposure from tobacco, and increase cessation efforts. PMID- 15825979 TI - Ethnic differences in trends and determinants of cigarette smoking in Hawaii. AB - BACKGROUND: The prevalence of smoking continues to be higher in some ethnic groups than in others, despite the overall decrease of smoking during the last 30 years. The goal of this project was to investigate the trends and predictors of cigarette smoking among a population of Caucasians, Japanese, Chinese, Filipino, and Native Hawaiians. METHODS: We combined data from 19 earlier studies conducted in Hawaii between 1975 and 2001. After excluding 1,885 individuals without smoking data, 158,629 subjects (40.0% Japanese, 30.3% Caucasian, 14.3% Hawaiian, 8.1% Filipino, 3.0% Chinese) were included in the analysis. The prevalence of current smoking and past smoking was estimated for 5-year periods by sex and ethnicity and age-adjusted to the state's population. We calculated odds ratios and 95% confidence intervals by using polytomous logistic regression to explore determinants of smoking, while controlling for clustering by study. RESULTS: Men were more likely to smoke than women throughout the study period, but they experienced a greater decline in smoking until 1994. We observed a small increase in smoking prevalence thereafter. Native Hawaiians reported the highest smoking prevalence, Japanese the lowest, and Caucasians intermediate levels. Graduate level education had the strongest inverse association with current and past smoking. Older age at interview, being married, and a higher body weight were related to lower smoking prevalence. CONCLUSIONS: Our data suggest a modest reversal in the declining smoking prevalence during the late 1990s. The persistent ethnic differences require new approaches that reach those groups who remain at high risk for adverse health effects from smoking. PMID- 15825980 TI - On the coding and reporting of race and ethnicity in New Hampshire for purposes of cancer reporting. AB - BACKGROUND: The objective was to investigate how data on race and ethnicity are collected by hospitals reporting to the New Hampshire State Cancer Registry (NHSCR). METHOD: NHSCR surveyed hospitals asking how information on race and ethnicity were collected. A review of relevant legal mandates and national guidelines was undertaken. RESULTS: Many hospitals lack policies on collection, computer systems fail to support national guidelines, and staff rely on visual inspection. CONCLUSIONS: Hospital staffs are not now culturally equipped to collect race and ethnicity in a meaningful way. The numerator in cancer incidence rates is most likely not accurate and for some smaller populations very biased. A new framework is needed that takes into account the needs of the democracy. PMID- 15825981 TI - Disparities in the coverage of cancer information in ethnic minority and mainstream mass print media. AB - BACKGROUND: Significant disparities in cancer mortality exist as a function of ethnicity and race in North America. Little is known, however, about the presentation of cancer information in mass media that targets ethnic minority groups. OBJECTIVES: 1) To evaluate the volume and type of cancer coverage and the readability of cancer articles in Canadian mainstream and ethnic minority newspapers; and 2) to compare newspaper coverage of cancer with Canadian cancer mortality. DESIGN: Seven mainstream and 25 ethnic minority (Jewish, First Nations, Black/Caribbean, East Indian) English-language newspapers were assessed for cancer coverage in the year 2000. Articles were analyzed by using frequencies and nonparametric tests. The total number of cancer articles (N=171) in ethnic minority papers and a random 20% from mainstream papers were also evaluated for readability level by using SMOG. RESULTS: There were a total of 748 cancer articles (721 mainstream; 27 ethnic). Coverage was weighted towards breast cancer (20.1% mainstream, 33.3% ethnic of cancer articles) and contained little or no coverage of prostate (7.4% mainstream, 8.6% ethnic), colorectal (3.9% mainstream, 3.7% ethnic), or lung (3.9% mainstream, 0 ethnic) cancers. The mean SMOG readability scores were Grades 12.7 and 13.2 for mainstream and ethnic papers, respectively. Readability scores differed significantly in ethnic newspapers, with the most difficult (highest readability) levels in East Indian (Grade 16.3) and the easiest (lowest readability) levels in First Nations (Grade 11.3) papers. Cancer articles were not highly culturally tailored, as measured by identification of specific ethnic minority groups within ethnic and mainstream newspapers. CONCLUSIONS: Cancer coverage in ethnic and mainstream newspapers did not accurately reflect the leading causes of cancer death in Canada. Results also suggest the need for the collection of cancer data by ethnic minority group in Canada. Without the disaggregation of cancer statistics by ethnicity, we cannot inform high-risk subgroups of the population and appropriately tailor cancer prevention and treatment programs. PMID- 15825982 TI - For the patient. Managing diabetes with cultural sensitivity and self empowerment. PMID- 15825983 TI - For the patient. Many nursing home patients do not receive diabetes medicine. PMID- 15825985 TI - For the patient. Understanding reasons for obesity is important to weight control programs. PMID- 15825984 TI - For the patient. Helping American Samoans quit smoking. PMID- 15825986 TI - For the patient. How chronic pain affects African Americans. PMID- 15825987 TI - For the patient. Collecting cancer information in New Hampshire. PMID- 15825988 TI - For the patient. Exercise is important to preventing and controlling type 2 diabetes. PMID- 15825989 TI - For the patient. Do Canadian newspapers print readable, timely cancer stories? PMID- 15825990 TI - [Epidemiologic surveillance of thyroid cancers]. PMID- 15825991 TI - [Stomach cancer]. AB - Despite its decline in incidence in developed countries, gastric cancer is the second digestive cancer in France and remains the second most cause of cancer related deaths in the world. Gastritis induced by H. pylori infection and food regimen are the most frequent precancerous gastric factors. However there is no definite clinical evidence of the benefit of eradication on cancer risk. By waiting for effective anti-H. pylorivaccine, H. pylori should be only eradicated in selected patients at the highest risk of cancer. The stake is to develop inexpensive tests for identification of individuals at high risk, depending on genotypic polymorphisms of both the bacterium and the host. The endoscopic diagnosis is currently made at an advanced stage, related to non-specific and late symptoms. The prognosis remains poor with 5 years overall survival rate less than 20%. Surgical resection with D1 lymphadenectomy is the gold standard curative treatment. An adjuvant therapy with chemoradiotherapy should be considered for patients at high risk for recurrence. Gastric cancer is considered to be a chemotherapy sensitive disease, but polychemotherapy regimens (fluorouraci +/- cisplatin +/- epirubicin) result in modest increased survival (median 9 months); yet, the promising effectiveness of new drugs (irinotecan, docetaxel, oxaliplatin, capecitabin, targeted biotherapies) makes us hope an improvement of results. A number of entities, linitis plastica, gastric MALT lymphoma and stromal tumors should be recognised, because must have a different treatment. PMID- 15825992 TI - [Horton's disease]. PMID- 15825993 TI - [Pathology of the thyroid: an exposed endocrine gland]. PMID- 15825994 TI - [Thyroid nodule]. AB - The thyroid nodule is a frequent, most often benign, chronic, multifocal and slowly progressive disease. The first line strategy is to diagnose cancerous nodules (<5%) and relies upon fine needle aspiration (FNA), a specialised technique which in trained hands has a false negative rate of below 5%. The interest to explore small thyroid nodules is controversial since the prognosis of thyroid cancer is excellent for lesions measuring less than 20 mm. Though imaging accuracy is quite limited in assessing the diagnosis of thyroid cancer, both ultrasounds (US) and thyroid scan are helpful to enhance nodular identification (>30%), to sort the nodules relevant for cytological sampling and to optimize the follow-up, the major source of health costs. Suspicious and non contributive FNAs must have a control FNA within 6 months. Nodules with a non suspicious FNA (>85%) require long term follow-up. This follow-up is mainly morphological. New or evolutive nodules, as assessed by palpation or US, will require iterative FNAs or should be considered for surgery. In patients with hyperfunctioning nodules on the scan (10 to 20%), a yearly evaluation of the TSH level is sufficient. These nodules account either for autonomously functioning ones, which slowly develop towards thyrotoxicosis, or for hyperplastic nodules frequently disclosing a lymphocytic thyroiditis. Morbidity due to thyroid autonomy is still underestimated especially in aging patients with TSH levels < or =0.60 mU/L. An algorithmic approach to the diagnostic and follow-up evaluation of thyroid nodule is suggested. PMID- 15825995 TI - [Hyperthyroidism: current concepts and management]. AB - Hyperthyroidism leads to an overproduction of thyroid hormones, which determine diverse expressions of thyrotoxicosis. However a high prevalence of mild and subclinical forms of hyperthyroidism are now recognised, since the biological measurement of thyroid hormones and TSH modified the diagnostic criteria. Clinical, biological, ultrasonographic and scintigraphic data allow the identification of the different causes of hyperthyroidism. This point is very important to assess the prognosis and to choose the right therapy. Graves' disease, solitary and multinodular toxic goiters constitute the main causes of hyperthyroidism, with iatrogenic thyrotoxicosis. Antithyroid drugs, surgery and radioactive iodine, are useful for the traditional forms of hyperthyroidism. But the choice of the therapy and the procedures for the follow-up has now a more rational background. PMID- 15825996 TI - [Current views on hypothyroidism in adults]. AB - The various aspects of primary thyroid insufficiency are presented. Subclinical hypothyroidism is a special condition the prevalence of which is increasing. In the face of a growing concern of the population, subclinical hypothyroidism receives much attention from endocrinologists and internists but despite recent recommendations therapeutic strategies are not clear enough yet. The question of the feasibility of the systematic screening, at least in predisposed people, for hypothyroidism is under study. Overt hypothyroidism which is usually preceded by a long phase of subclinical hypothyroidism is therefore presentable if substitutive treatment is given early enough. Occasional patients, however, may rich the level of profound hypothyroidism unnoticed. The modalities of treatment of over hypothyroidism are described. Latrogenic hypothyroidism is also described. Finally, myxoedema coma is discussed not because its prevalence, which is very low, but because this condition may be forgotten. PMID- 15825997 TI - [Multinodular goiters]. AB - The incidence of multinodular goiters (MNG) differs according to the countries, and seems to be widely dependent of the iodine status. In France, the SU.VI.MAX study indicates that 4.75% of the active population is suffering from MNG, this incidence is increased with gender and age. The role of genetic factors is estimated as being around 80%. The remaining 20% are related to environmental factors, some of them being still unknown. Admit these external factors, the iodine deficiency is predominant and should be theoretically easy to avoid. Most of MNG develop on diffuse goiters and could be prevented when these underlying lesions are treated early by the administration of thyroid hormone alone or associated with iodide potassium. Once the nodularity of the thyroid tissue is present, the medical treatment is of weak efficacy, it usually stabilises the goiter and prevents the occurrence of new nodules. The two main treatments are surgery and radioactive iodine, the latter is currently used not only when the former is contra-indicated. In some countries, radioactive iodine is considered as the best alternative in the majority of cases. PMID- 15825998 TI - [Thyroid dysfunction and pregnancy]. AB - The pregnancy is a state of thyroid hyperstimulation, therefore of changes of thyroid hormone values. Results of thyroid hormone measurement have to be analysed in the context of gestation age. Hyperthyroidism, mostly represented by Graves' disease, requires a multidisciplinary management, owing to possible maternal, foetal and neonatal complications. Treatment with antithyroid drugs, is a compromise between the risk of uncontrolled maternal hyperthyroidism and the risk of iatrogenic foetal hypothyroidism. Evaluation of foetal thyroid function considers the titre of thyrotropin receptor antibodies in the mother's blood, the dose of antithyroid drugs to maintain euthyroidism in the mother, and the signs of foetal hyperthyroidism on ultrasound. Maternal hypothyroidism is associated with foetal and maternal morbidity. Untreated or inappropriately treated, it is associated with poorer performances of offspring in intelligence tests. Thyroid autoimmunity is associated with hypofertility, particularly with spontaneous abortion. Screening for thyroid dysfunction during pregnancy, although not systematic, should have broad indications. PMID- 15825999 TI - [Iodine excess and thyroid dysfunction]. AB - Hypothyroidism or hyperthyroidism may develop during iodine excess. Hypothyroidism occurs mostly in newborns or in elderly patients with underlying autoimmune thyroiditis and is treated by substitutive doses of thyroxine. Hyperthyroidism is of two types. Type I develops in patients with pretoxic thyroid glands, the radioactive tracer uptake remains high. This type is treated with thionamides and potassium perchlorate, as well as, in some cases, by thyroidectomy or iodine 131. Type II develops in non pretoxic thyroid gland, and it evoluates spontaneously towards euthyroidism, in several months, despite continuation of iodine excess. It responds to glucocorticoid therapy. PMID- 15826000 TI - [Graves' ophthalmopathy]. AB - Graves' ophthalmopathy (GO) is a chronic autoimmune process that affects the retrobulbar tissue and has strong etiological links with graves' disease. Pathogenesis is incompletely understood. Symptoms include proptosis, extraocular muscle dysfunction, eyelid swelling and retraction. In its severe expression, it is a disfiguring and potentially sight threatening disease that profoundly influences and impairs the quality of life of affected individuals. Only a minority of patients develop severe expressions of the disease, requiring aggressive treatments. Glucocorticoids, orbital radiotherapy and decompressive orbital surgery represent a mainstay and a well established treatment for severe and active GO. PMID- 15826001 TI - [Does thyroid scintigram still have indications in 2005?]. AB - Since 15 years, the indications of thyroid scintigrams have been dramatically reduced. This examination appeared useless in most cases of hypothyroidism and for the evaluation of thyroid cysts or for infracentimetric nodules. For greater solid nodules, its poor sensibility and bad specificity have resulted in its replacement by echography and fine needle biopsy. Thyroid scintigram is however usefull for the non obvious etiologic diagnosis of thyrotoxicosis and in case of thyroid goiter with a low TSH and normal thyroid hormons. Whole body131 I scans remain usefull for thyroid cancer follow up, especially after therapeutic dose of radioiodine. PMID- 15826002 TI - [Thyroid diseases: what you need to know]. PMID- 15826003 TI - [Old and new reforms]. PMID- 15826004 TI - [Oral pain]. PMID- 15826005 TI - [Pathology of inoculation]. PMID- 15826006 TI - [Iatrogenic disease]. PMID- 15826007 TI - [The sick elderly person: symptomatology, psychological and therapeutic aspects]. PMID- 15826008 TI - [History of Louis XIV's disease in Calais]. PMID- 15826010 TI - Chemistry and properties of nanocrystals of different shapes. PMID- 15826012 TI - New approaches to nanofabrication: molding, printing, and other techniques. PMID- 15826011 TI - Self-assembled monolayers of thiolates on metals as a form of nanotechnology. PMID- 15826013 TI - Synthesis and properties of molecular rods. 2. Zig-zag rods. PMID- 15826014 TI - Artificial molecular rotors. PMID- 15826015 TI - Toward intelligent molecular machines: directed motions of biological and artificial molecules and assemblies. PMID- 15826016 TI - Supramolecular nanotube architectures based on amphiphilic molecules. PMID- 15826017 TI - Self-assembled nanoreactors. PMID- 15826018 TI - About supramolecular assemblies of pi-conjugated systems. PMID- 15826019 TI - Nanostructures in biodiagnostics. PMID- 15826020 TI - Phenolic acids enzymatic lipophilization. AB - Lipophilization is the esterification of a lipophilic moiety (fatty acid or fatty alcohol) on different substrates (phenolic acid, sugar, protein, ...), resulting in new molecules with modified hydrophilic/lipophilic balance. This reaction can be obtained chemically or enzymatically using different enzymes. Phenolic acids possess interesting biological properties (antioxidant, chelator, free radical scavenger, UV filter, antimicrobial, ...), but because of their relatively low solubility in aprotic media, their application in oil-based products is limited. Therefore, the esterification of their carboxylic acid function with a fatty alcohol enhances their hydrophobicity and results in a multifunctional amphiphilic molecule. Enzymatic lipophilization of phenolic acids is nowadays studied for potential industrial applications. Different systems have been proposed to perform the reaction yield [free or immobilized enzymes (lipase, feruloyl esterase, tannase, etc.), free or added organic solvent, addition of surfactant, microemulsion system, etc.]. Some of the functional properties of these esters have been demonstrated. This review presents a panorama of the advances in this field. PMID- 15826021 TI - The physics and chemistry behind the bubbling properties of champagne and sparkling wines: a state-of-the-art review. AB - In this review, the latest results about the chemical physics behind the bubbling properties of Champagne and sparkling wines are collected and fully illustrated. The chemistry of carbon dioxide molecules dissolved into the liquid matrix (section 2) is presented, as are the three main steps of a fleeting bubble's life, that is, the bubble nucleation on tiny particles stuck on the glass wall (section 3), the bubble ascent and growth through the liquid matrix (section 4), and the bursting of bubbles at the liquid surface (section 5), which constitutes the most intriguing, functional, and visually appealing step. PMID- 15826022 TI - Investigation of the potential utility of single-bounce attenuated total reflectance Fourier transform infrared spectroscopy in the analysis of distilled liquors and wines. AB - A new Fourier transform infrared (FTIR) spectroscopic method based on single bounce attenuated total reflectance (SB-ATR) spectroscopy was developed for the analysis of distilled liquors and wines. For distilled liquors, a partial least squares (PLS) calibration was developed for alcohol determination based on the SB ATR/FTIR spectra of mixtures of ethanol and distilled water. An independent set of 12 different distilled liquor samples was predicted from the PLS calibration, and a standard deviation of the differences for accuracy (SDD(a)) between actual and predicted values of 0.142% (v/v) was obtained. The potential utility of SB ATR/FTIR spectroscopy for the analysis of wines was initially evaluated based on a comparison with Fourier transform near-infrared (FT-NIR) spectroscopy and FTIR spectroscopy using a flow-through transmission cell. PLS calibrations for alcohol, total reducing sugars, total acidity and pH were developed using pre analyzed wine samples (n = 28), and for SB-ATR/FTIR spectroscopy, the SDD(a) for the leave-one-out cross-validation statistics were of the order of 0.100% (v/v), 0.707 g L(-1), 0.189 g L(-1) (H2SO4), and 0.230, respectively. Overall, the SB ATR/FTIR results were better than those obtained using FT-NIR spectroscopy and comparable to those obtained with transmission FTIR spectroscopy. A PLS calibration based on preanalyzed wine samples (n = 72) for the prediction of 11 different components and parameters in wines by SB-ATR/FTIR spectroscopy was subsequently developed and validated using an independent sample set (n = 77). Good coefficients of correlation between the reference and predicted values for the validation set were obtained for most of the components and parameters except citric acid, volatile acids, and total SO2. The results of this study demonstrate the suitability of SB-ATR/FTIR spectroscopy for the routine analysis of distilled liquors and wines. PMID- 15826023 TI - Detection of extra virgin olive oil adulteration with lampante olive oil and refined olive oil using nuclear magnetic resonance spectroscopy and multivariate statistical analysis. AB - High-field 31P NMR (202.2 MHz) spectroscopy was applied to the analysis of 59 samples from three grades of olive oils, 34 extra virgin olive oils from various regions of Greece, and from different olive varieties, namely, 13 samples of refined olive oils and 12 samples of lampante olive oils. Classification of the three grades of olive oils was achieved by two multivariate statistical methods applied to five variables, the latter being determined upon analysis of the respective 31P NMR spectra and selected on the basis of one-way ANOVA. The hierarchical clustering statistical procedure was able to classify in a satisfactory manner the three olive oil groups. Subsequent application of discriminant analysis to the five selected variables of oils allowed the grouping of 59 samples according to their quality with no error. Different artificial mixtures of extra virgin olive oil-refined olive oil and extra virgin olive oil lampante olive oil were prepared and analyzed by 31P NMR spectroscopy. Subsequent discriminant analysis of the data allowed detection of extra virgin olive oil adulteration as low as 5% w/w for refined and lampante olive oils. Further application of the classification/prediction model allowed the estimation of the percent concentration of refined olive oil in six commercial blended olive oils composed of refined and virgin olive oils purchased from supermarkets. PMID- 15826024 TI - Analysis of leaf surface sesquiterpenes in potato varieties. AB - A comparative study of potato leaf sesquiterpenes was carried out. GC, GC-MS, and NMR analyses were used to identify and quantify the sesquiterpenes in the leaf surfaces of 10 potato (Solanum tuberosum) varieties. Two sesquiterpene alcohols and 17 sesquiterpene hydrocarbons were identified and quantitatively determined. The distribution of the sesquiterpenes was found to be variety-specific. The sesquiterpene contents of the different potato varieties were subjected to cluster and principal component analyses. The eight potato varieties of the main chemotype cluster were dominated by beta-caryophyllene (9-148 ng/cm2), germacrene D (2-46 ng/cm2), germacrene D-4-ol (0.4-31 ng/cm2), beta-sesquiphellandrene (1-34 ng/cm2), and an unknown sesquiterpene alcohol III (0.2-37 ng/cm2). Chemometric classification distinguished two varieties, Mila and Vistula, from a major cluster. The Vistula variety was distinguished from the others by its high contents of beta-caryophyllene, alpha-humulene, germacrene D, and germacrene D-4 ol and the Mila variety by beta-elemene, trans-alpha-bergamotene, (Z)-beta farnesene, (E)-beta-farnesene, trans-beta-bergamotene, beta-sesquiphellandrene, and unknown sesquiterpene alcohols I, II, III. PMID- 15826025 TI - Differentiation of carbohydrate gums and mixtures using fourier transform infrared spectroscopy and chemometrics. AB - Guar gum, a nonionic galactomannan, is used as an economical thickener and stabilizer in the food industry and is often combined with xanthan, locust bean gum (LBG), or carboxymethylcellulose (CMC) to promote synergistic changes in viscosity or gelling behavior via intermolecular interactions; however, the adulteration of LBG with guar gum is a well-known industrial problem. The ability to identify the purity of gums and concentrations of individual gums in mixtures would be advantageous for quality control in the food industry. Fourier transform infrared spectroscopy (FTIR) methods are rapid and require minimum sample preparation. The objectives of this study were to evaluate the ability of FTIR techniques to (1) differentiate LBG with a variety of mannose/galactose (M/G) ratios, (2) differentiate guar, LBG, tara, and fenugreek gums, (3) differentiate pure guar gum from guar gum mixed with LBG, xanthan gum, or CMC, (4) quantify LBG, xanthan gum, and CMC in guar gum, and (5) quantify guar gum in LBG. Two FTIR methods were used: diffuse reflectance (DRIFT) on powdered gum samples added to KBr at 5%, w/w, and attenuated total reflectance (ATR) on 1%, w/w, gum solutions. Spectra were collected and then analyzed by multivariate statistical procedures (chemometrics). The DRIFT method provided better discrimination and quantitative results than the ATR method. Canonical variate analysis (CVA) of DRIFT spectra (1200-700 cm(-1)) was able to classify LBG with various M/G ratios, pure galactomannans, and pure versus mixtures of gums with 100% accuracy. Quantification of an individual gum in gum mixtures (0.5-15%, w/w) was possible using partial least-squares (PLS) analysis of DRIFT spectra with R2 > 0.93 and using this approach for quantifying guar gum added to LBG resulted in an R2 > 0.99, RMSEC = 0.29, and RMSEP = 3.31. Therefore, the DRIFT FTIR method could be a useful analytical tool for quality control of select gums and gum mixtures used in the food industry. PMID- 15826026 TI - Identification and quantification of epsilon-(gamma-glutamyl)lysine in digests of enzymatically cross-linked leguminous proteins by high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). AB - A rapid and convenient method for the precise quantification of epsilon-(gamma glutamyl)lysine isopeptide in lyophilized proteolytic digests of cross-linked plant protein samples was developed. The isopeptide was baseline-separated from three other isomers containing lysyl and glutamyl residues by reverse-phase high performance liquid chromatography after exhaustive proteolytic digestion of the samples cross-linked by a microbial transglutaminase (MTG). Highly selective detection was performed by electrospray mass spectrometry in MS/MS mode. Demonstrating the applicability of the suggested analytical procedure, enzymatic cross-linking of protein isolates from soy [Glycine max (L.) Merr.], pea [Pisum sativum L.], and the sweet lupin species Lupinus albus L. and Lupinus angustifolius L. was investigated after incubation with 0.01 g of MTG/100 g of protein for 0-240 min at 40 degrees C. The liquid chromatography-mass spectrometry (LC-MS) method was successfully applied to monitor the kinetics of epsilon-(gamma-glutamyl)lysine isopeptide formation. Since the calculated initial levels of epsilon-(gamma-glutamyl)lysine in the genuine leguminous protein isolates were between 40 and 77 micromol/100 g, an isopeptide detection limit of 0.5 microg/mL, corresponding to approximately 50 micromol/100 g of protein, was shown to suffice for quantifying the cross-linking rate enzymatically induced by MTG. Concentrations of epsilon-(gamma-glutamyl)lysine in the texturized proteins ranged from 100 to 500 micromol/100 g of protein. PMID- 15826027 TI - Determination of major carotenoids in a few Indian leafy vegetables by high performance liquid chromatography. AB - Leafy vegetables [Basella rubra L., Peucedanum sowa Roxb., Moringa oleifera Lam., Trigonella foenum-graecum L., Spinacia oleracea L., Sesbania grandiflora (L.) Poir., and Raphanus sativus L.] that are commonly used by the rural population in India were evaluated in terms of their main carotenoid pattern. The extracted carotenoids were purified by open column chromatography (OCC) on a neutral alumina column to verify their identity by their characteristic UV-visible absorption spectra. Reverse-phase high-performance liquid chromatography (HPLC) on a C18 column with UV-visible photodiode array detection under isocratic conditions was used for quantification of isolated carotenoids. Acetonitrile/methanol/dichloromethane (60:20:20 v/v/v) containing 0.1% ammonium acetate was used as a mobile phase. The major carotenoids identified by both methods were lutein, beta-carotene, violaxanthin, neoxanthin, and zeaxanthin. Among the carotenoids identified, lutein and beta-carotene levels were found to be higher in these leafy vegetables. Results show that P. sowa and S. oleracea are rich sources of lutein (77-92 mg/100 g of dry wt) and beta-carotene (36-44 mg/100 g of dry wt) compared with other leafy vegetables. The purity of carotenoids eluted by OCC was clarified by HPLC, and they were found to be 92% +/ 3% for neoxanthin, 94% +/- 2% for violaxanthin, 97% +/-2% for lutein and zeaxanthin, and 90% +/- 3% for beta-carotene. It could be recommended to use P. sowa and S. oleracea as rich sources of lutein and beta-carotene for health benefits. The OCC method proposed is relatively simple and provides purified carotenoids for feeding trials. PMID- 15826028 TI - Rapid prediction of acid detergent fiber, neutral detergent fiber, and acid detergent lignin of rice materials by near-infrared spectroscopy. AB - A rapid predictive method based on near-infrared spectroscopy (NIRS) was developed to measure acid detergent fiber (ADF), neutral detergent fiber (NDF), and acid detergent lignin (ADL) of rice stem materials. A total of 207 samples were divided into two subsets, one subset (approximately 136 samples) for calibration and cross-validation and the other subset for independent external validation to evaluate the calibration equations. Different mathematical treatments were applied to obtain the best calibration and validation results. The highest coefficient of determination for calibration (R2) and coefficient of determination for cross-validation (1-VR) were 0.968 and 0.949 for ADF, 0.846 and 0.812 for NDF, and 0.897 and 0.843 for ADL, respectively. Independent external validation still gave a high coefficient of determination for external validation (r2) and a low standard error of performance (SEP) for the three parameters; the best validation results were SEP = 0.933 and r2 = 0.959 for ADF, SEP = 2.228 and r2 = 0.775 for NDF, and SEP = 0.616 and r2 = 0.847 for ADL, indicating that NIR gave a sufficiently accurate prediction of ADF and ADL content of rice material but a less satisfactory prediction for NDF. This study suggested that routine screening for these forage quality parameters with large numbers of samples is possible with NIRS in early-generation selection in rice-breeding programs. PMID- 15826029 TI - Study of enrofloxacin depletion in the eggs of laying hens using diphasic dialysis extraction/purification and determinative HPLC-MS analysis. AB - A study of the depletion of enrofloxacin residues in eggs was carried out using a diphasic dialysis procedure for the extraction of fluoroquinolone residues from the matrix. Enrofloxacin was administered to laying hens through the intramuscular route (15 mg/day) and orally (12 mg/day). After daily collection, the egg albumen and the egg yolk were separated, and the residue levels were determined using an HPLC-MS (API-ESI) method. The enrofloxacin and ciprofloxacin peaks gradually increased until the fifth day, because the drug was employed for 5 days. However, differences were observed in the depletion curves of enrofloxacin and its metabolite when both parts of the egg and the mode of administration were considered. PMID- 15826030 TI - Phenylpropanoid glycosides from Tynanthus panurensis: characterization and LC-MS quantitative analysis. AB - A phytochemical analysis of the methanol extract of Tynanthus panurensis bark led to the isolation of one new phenylpropanoid glycoside, eugenol-O-[beta-D xylopyranosyl-(1-->5)-O-beta-D-apiofuranosyl-(1-->6)-O-beta-D-glucopyranoside], the known verbascoside, isoverbascoside, and leucosceptoside, along with the known flavonoid apigenin 8-C-[beta-D-xylopyranosyl-(1-->6)-beta-D glucopyranoside], namely, katchimoside. Their structures were established by NMR and ESIMS experiments. Additionally, a quantitative study of the phenylpropanoid glycosides fraction of T. panurensis bark and of the hydroalcoholic extract prepared according to the traditional recipe was performed by combining high performance liquid chromatography diode array detection with positive electrospray ionization tandem mass spectrometry. The new eugenol derivate was found to be the most abundant phenylpropanoid glycoside in both dried bark (19.5 mg/g) and hydroalcoholic extract (0.24 mg/mL). The antioxidant activity of all the isolated compounds and of the methanol and hydroalcoholic extract of the bark was determined by measuring the free radical scavenging effects using the Trolox equivalent antioxidant capacity method. The traditional hydroalcoholic extract showed a moderate activity. PMID- 15826031 TI - Analysis of anthocyanins in rat intestinal contents--impact of anthocyanin chemical structure on fecal excretion. AB - Absorption of dietary anthocyanins is limited; however, fecal anthocyanin excretion has been rarely studied. We developed a method for extraction and analysis of fecal anthocyanins. Aqueous methanol (60%) maximized extraction efficiency (approximately 88%). Severe anthocyanin degradation (monitored by high performance liquid chromatography) was observed in feces stored at -18 degrees C; therefore, storage time should be minimized and lower temperatures used. Fecal and cecal content samples were collected from 32 rats receiving either chokeberry, bilberry, grape-enriched (3.85 g monomeric anthocyanin per kg diet), or control diet for 14 weeks. Fecal anthocyanin concentrations were significantly different among groups (0.7/1.8/2.0 g/kg wet feces, chokeberry/bilberry/grape). Anthocyanin profiles of cecal contents and feces were similar. Losses in the intestinal contents were high for anthocyanin glucosides, moderate for galactosides, and negligible for arabinosides or xylosides. Acylation or diglucosylation enhanced anthocyanin stability. High anthocyanin concentration in the fecal content may favor anthocyanin absorption into the colon epithelial cells, resulting in potential health benefits. PMID- 15826032 TI - Uptake of thallium from artificially and naturally contaminated soils into rape (Brassica napus L.). AB - An inductively coupled plasma mass spectrometry (ICP/MS) method was used for the evaluation of thallium transfer from naturally (pedogeochemically) and artificially contaminated soils into rape. Two sets of three different types of top soils (heavy, medium, and light) were used for pot experiments. The first set was collected from areas with high levels of pedogeochemical thallium (0.3, 1.5, and 3.3 mg kg(-1) DM). The second set of three soils with naturally low content of thallium was artificially contaminated with thallium sulfate to achieve five levels of contamination (0, 0.4, 2, 4, and 6 mg kg(-1) DM Tl). The soil samples and the samples of winter and spring rape (straw, seeds) from both sets were collected and analyzed. Plant and soil samples from fields were collected at 42 selected sites situated in South Bohemia and in Czech-Moravian Highlands where higher pedogeochemical content of thallium was expected. More intensive transport (better availability) of Tl was observed in the case of artificially contaminated soils. The physicochemical form and the total content of Tl in soil were found to be the main factors influencing its uptake by plants. The concentration of Tl in rapeseeds in the field samplings was mostly 45% of its content in the particular soil. Nevertheless the uptake of Tl from soils with naturally high pedogeochemical content can be high enough to seriously endanger food chains. These findings are very important because of the high toxicity of Tl and the absence of threshold limits for Tl in soils, agricultural products, feedstuffs, and foodstuffs in most countries including the Czech Republic. PMID- 15826033 TI - Application of cluster analysis (CLA) in feed chemical imaging to accurately reveal structural-chemical features of feeds and plants within cellular dimension. AB - Synchrotron Fourier transform infrared (FTIR) microspectroscopy can explore molecular chemical features of the microstructure of feeds. The most straightforward method of data analysis is the mapping of specific functional group intensities and frequencies by peak heights and/or areas. However, this univariate statistical method does not always accurately identify functional group locations and concentrations, because the so-called "unique" bands for the peak area mapping have more or less inference with other nonunique bands in feed and plant tissues. The objective of this study was to use a multivariate analysis method--called agglomerative hierarchical cluster analysis (CLA)--to analyze infrared spectra for chemical imaging. The results show the CLA method gave satisfactory results and was conclusive in showing that it can discriminate and classify functional group differences existing in different structure regions. This approach (CLA for chemical imaging) places synchrotron FTIR microspectroscopy at the forefront of those techniques that could potentially be used in the rapid characterization of feed microstructure. PMID- 15826034 TI - Development of a radioimmunoassay for the quantitative determination of 8 prenylnaringenin in biological matrices. AB - Seven carboxylic acid haptens of 8-prenylnaringenin (8-PN) were synthesized, coupled to cationized bovine serum albumin, and employed to raise specific antisera in rabbits. Two linkers of different lengths (C3H6COOH and C6H12COOH) were coupled to the C7-OH group and separated into their respective enantiomers yielding the first four haptens. Racemic derivatives with C4'-OH coupled linkers C5H10COOH and C9H18COOH were synthesized carrying a methylated C7-OH. Another racemic C4'-OH hapten (CH2COOH) was prepared starting from naringenin. The haptens elicited variable antibody titers dependent on linker lengths, with short linkers giving the best results. Three antisera were characterized in detail: anti-C7-carboxy-propyloxy-2S-(-)-8-PN (anti-H-11), anti-C7-carboxy-propyloxy-2R (+)-8-PN (anti-H-10), and anti-C4'-carboxy-methoxy-rac-8-PN (anti-H-25). anti-H 10 and anti-H-11 showed about 9% enantiomeric cross-reactivity, and anti-H-11 did not discriminate between isoxanthohumol (IX) and 8-PN (84% cross-reactivity). For anti-H-10, cross-reactivities in the range of 2-5% were found for xanthohumol, IX, and 6-prenylnaringenin. Respective numbers for anti-H-25 were 0.02, 0.1, and 0.2%. Tritiated 8-PN was synthesized yielding a 3H-tracer of high specific radioactivity (2.22 GBq/mg). A radioimmunoassay using anti-H-25 and 3H-8-PN was established and used for the quantitative determination of 8-PN in various beer brands and in the urine of six men after the consumption of three different brands of beer. Furthermore, the dose-dependent excretion of 8-PN was tested after the consumption of a higher volume of a single beer brand with and without spiking with 8-PN and a small oral dose of authentic 8-PN, respectively. Conflicting results led to a pilot test on the in vivo conversion (demethylation) of IX into 8-PN in two men. Conversion rates of 1.9 and 4.4% were estimated. Thus, the total 8-PN dose in beer brands spiced with natural hop or hop products seems to be the sum of the 8-PN amount in a consumed volume and the amount arising from the conversion of IX. PMID- 15826035 TI - Interaction between morin and sodium dodecyl sulfate (SDS) micelles. AB - Electronic absorption spectra, fluorescence emission spectra, ATR-FTIR spectra, cyclic voltammetric measurements, and ab initio quantum calculation are used to study the properties of morin in SDS micelles of different microstructures and microenvironments and to gain the information about the binding of morin with the SDS micelles. Morin can be located in the SDS micelles with its phenyl group (deviating by 38.98 degrees from the planarity), which leads to the increase of the planarity and the extension of pi conjugation of the whole molecule. The embedment of two hydroxyl groups on the phenyl into a more hydrophobic microenvironment makes the oxidation peak of morin move to a higher potential with a decreased peak current. The binding constant (K) and the distribution coefficient (P) of morin in the spherical SDS micelle are larger than those in the rodlike SDS micelle. The binding of morin with SDS micelle is a spontaneous (DeltaG < 0) and exothermic process (DeltaH < 0), and the hydrophobic force is the main driving force for its solubilization. PMID- 15826036 TI - Increased salicylate concentrations in urine of human volunteers after consumption of cranberry juice. AB - The aim of this study was to assess whether regular consumption of cranberry juice results in elevations in urinary salicylate concentrations in persons not taking salicylate drugs. Two groups of healthy female subjects (11/group) matched for age, weight, and height consumed 250 mL of either cranberry juice or a placebo solution three times a day (i.e., 750 mL/day) for 2 weeks. At weekly intervals, salicylic acid and salicyluric acid (the major urinary metabolite of salicylic acid) concentrations were determined in urine by HPLC with electrochemical detection. Concentrations of salicylic acid in plasma were also determined. Consumption of cranberry juice was associated with a marked increase (p < 0.001) of salicyluric and salicylic acids in urine within 1 week of the intervention. After 2 weeks, there was also a small but significant (p < 0.05) increase in salicylic acid in plasma. The regular consumption of cranberry juice results in the increased absorption of salicylic acid, an anti-inflammatory compound that may benefit health. PMID- 15826037 TI - Phenolic compounds in external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC). AB - Glycosylated kaempferol derivatives from the external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC) characterized by reversed-phase HPLC-DAD MS/MS-ESI were kaempferol 3-O-sophorotrioside-7-O-glucoside, kaempferol 3-O- (methoxycaffeoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7 O-glucoside, kaempferol 3-O-sophorotrioside-7-O-sophoroside, kaempferol 3-O sophoroside-7-O-sophoroside, kaempferol 3-O-tetraglucoside-7-O-sophoroside, kaempferol 3-O-(sinapoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O (feruloyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside, kaempferol 3-O-(sinapoyl)sophoroside, kaempferol 3-O-(feruloyl)sophorotrioside, kaempferol 3-O-(feruloyl)sophoroside, kaempferol 3-O-sophoroside, and kaempferol 3-O-glucoside. These acylated derivatives are reported for the first time in nature, with the exception of kaempferol 3-O-(sinapoyl)sophoroside. Quantification of the identified compounds was achieved by HPLC-DAD and carried out in samples cultivated under conventional or organic practices and collected at different times. In general, samples from organic production exhibited higher total phenolics content than those from conventional practices collected in the same period. PMID- 15826038 TI - Andrastins A-D, Penicillium roqueforti Metabolites consistently produced in blue mold-ripened cheese. AB - This is the first finding of andrastins in blue cheese as well as any other sample type. Here, they were produced by the secondary starter culture Penicillium roqueforti. After purification by normal-phase chromatography followed by combined reverse-phase ion-exchange chromatography, the andrastins A D were detected by liquid chromatography combined with UV and high-resolution mass spectrometry. In 23 representative samples of European blue cheeses, andrastin A was consistently found in quantities between 0.1 and 3.7 microg/g of cheese (median 2.4 microg/g). Assuming the same molar response factors as for andrastin A, the B, C, and D analogues were present in approximately 5-, 3-, and 5-20-fold lower amounts than andrastin A, respectively. The andrastins are protein farnesyltransferase inhibitors and are capable of inhibiting the efflux of anticancer drugs from multidrug-resistant cancer cells. Thus, their presence in common blue cheese suggests a potential for a positive or negative impact on human health. PMID- 15826039 TI - In vitro investigation into the potential prebiotic activity of honey oligosaccharides. AB - The effect of honey oligosaccharides on the growth of fecal bacteria was studied using an in vitro fermentation system. Prior to treatment, glucose and fructose (31.73 and 21.41 g/100 g of product, respectively) present in honey, which would be digested in the upper gut, were removed to avoid any influence on bacterial populations in the fermentations. Nanofiltration, yeast (Saccharomyces cerevisiae) treatment, and adsorption onto activated charcoal were used to remove monosaccharides. Prebiotic (microbial fermentation) activities of the three honey oligosaccharide fractions and the honey sample were studied and compared with fructooligosaccharide (FOS), using 1% (w/v) fecal bacteria in an in vitro fermentation system (10 mg of carbohydrate, 1.0 mL of basal medium). A prebiotic index (PI) was calculated for each carbohydrate source. Honey oligosaccharides seem to present potential prebiotic activity (PI values between 3.38 and 4.24), increasing the populations of bifidobacteria and lactobacilli, although not to the levels of FOS (PI of 6.89). PMID- 15826040 TI - Antifungal synergistic effect of scopoletin, a hydroxycoumarin isolated from Melia azedarach L. fruits. AB - In the continuous search for antifungal compounds from plants, the hydroxycoumarin scopoletin (1) was isolated from seed kernels of Melia azedarach L. from which three other compounds, vanillin (2), 4-hydroxy-3 methoxycinnamaldehyde (3), and (+/-) pinoresinol (4), have also been isolated. Guided fractionation through autobiography on TLC using Fusarium verticillioides (Saccardo) Nirenberg as test organism led to the isolation of 1, which exhibited a minimum inhibitory concentration (MIC) of 1.50 mg/mL in the microbroth dilution method. Despite its own weak activity, when the coumarin was combined with the above-mentioned compounds, a strong enhancement of the antifungal effect was observed, even showing a complete inhibition in the growth of the pathogen when 1 was added at a concentration of up to 5% of its MIC value. The same level of effectiveness was observed when the synthetic antifungal agents Mancozeb and Carboxin were each combined with compounds 1-4, in which cases it became possible to decrease the effective concentrations of these commercial compounds by up to 2.5 and 3%, respectively. PMID- 15826041 TI - Antioxidant activity of dietary fruits, vegetables, and commercial frozen fruit pulps. AB - Fruits, vegetables, and commercial frozen pulps (FP) consumed in the Brazilian diet were analyzed for antioxidant activities using two different methods, one that determines the inhibition of copper-induced peroxidation of liposome and another based on the inhibition of the co-oxidation of linoleic acid and beta carotene. The anthocyanin-rich samples showed the highest, concentration dependent, antioxidant activities in both systems. In the liposome system, at both 10 and 50 microM gallic acid equivalent (GAE) addition levels, the neutral and acidic flavonoids of red cabbage, red lettuce, black bean, mulberry, Gala apple peel, jambolao, acai FP, mulberry FP, and the acidic flavonoids of acerola FP showed the highest antioxidant activities (>85% inhibition). In the beta carotene bleaching system, the samples cited above plus red guava gave inhibition values >70%. On the other hand, some samples showed pro-oxidant activity in the liposome system coincident with a low antioxidant activity in the beta-carotene system. There was no relationship between total phenolics content, vitamin C, and antioxidant activity, suggesting that the antioxidant activity is a result of a combination of different compounds having synergic and antagonistic effects. PMID- 15826042 TI - Determination of beta-carotene and lutein available from green leafy vegetables by an in vitro digestion and colonic fermentation method. AB - Green leafy vegetables (Spinacea oleracea, Cnidoscolus aconitifolius, and Solanum americanum) contain a high amount of beta-carotene (27-52 mg/100 g of dry sample) and lutein (140-193 mg/100 g of dry sample). The amount of beta-carotene and lutein released from the food matrix by the action of digestive enzymes ranged from 22 to 67% and from 27 to 77%, respectively. There was a significant correlation between the enzymatic release of carotenoids (lutein + beta-carotene) and the content of Klason lignin, nonstarch polysaccharides, and resistant protein. The carotenoids released by the in vitro colonic fermentation ranged from 2 to 11%, and part of them (0.251-4.03 mg/100 g of original dry sample) remained intact in the fermentation media and could be potentially absorbed in the colon. A significant part of carotenoids seems to be unavailable in the intestinal tract (16% in S. oleracea to 58% in C. aconitifolius). PMID- 15826043 TI - Triterpene glycosides of Siraitia grosvenori inhibit rat intestinal maltase and suppress the rise in blood glucose level after a single oral administration of maltose in rats. AB - The effect of the crude extract from Siraitia grosvenori Swingle (SG-ex) on the postprandial rise in blood glucose level was investigated. The increase in plasma glucose level in response to the oral administration of maltose was significantly suppressed in rats when SG-ex was given orally 3 min before the maltose administration. There was, however, no effect when glucose was administered instead, suggesting that the antihyperglycemic effect of SG-ex is elicited by inhibition of maltase in the small intestinal epithelium. In vitro, SG-ex inhibited rat small intestinal maltase. Similar effects were also observed both in vivo and in vitro when the concentrate of the sweet elements (triterpene glycosides) prepared from SG-ex was used. Furthermore, the main sweet element of SG-ex, mogroside V, and some minor elements such as mogroside IV, siamenoside I, and mogroside III also exhibited maltase inhibitory effect with IC50 values of 14, 12, 10, and 1.6 mM, respectively. These results suggest that SG-ex exerts anti-hyperglycemic effects in rats by inhibiting maltase activity and that these effects are at least partially exerted by its sweet elements, triterpene glycosides. PMID- 15826044 TI - Composition and antibacterial activity of Pseudocytisus integrifolius (Salisb.) essential oil from Algeria. AB - The essential oil composition of an endemic Algerian Cruciferae, Pseudocytisus integrifolius (Salisb.) Rehder, was analyzed by gas chromatography (GC) and GC mass spectrometry (MS). Eighty-three components representing more than 96.5% of the oil were identified. The major components were dimethyl disulfide (33.4%), dimethyl trisulfide (24.2%), and an unsaturated nitrile (31.7%). Fractionation on a silica gel column led to the identification of trace-level compounds, in particular, polar compounds such as nitriles and aldehydes, and to the isolation of dimethyl disulfide, dimethyl trisulfide, and an unsaturated nitrile. Structural analysis using high-resolution mass spectrometry (HRMS) and 1H,13C NMR techniques enabled the identification of pent-4-enenitrile. Variation in essential oil composition and yields was studied according to harvesting time, drying, and parts of the plant. The essential oil of aerial parts was tested for its antibacterial activity using a paper disk method. The oil was effective on the inactivation of Escherichia coli and Pseudomonas aeruginosa and ineffective on the inactivation of Staphylococcus aureus. PMID- 15826045 TI - Stability of ferric complexes with 3-hydroxyflavone (flavonol), 5,7 dihydroxyflavone (chrysin), and 3',4'-dihydroxyflavone. AB - The acid dissociation and ferric stability constants for complexation by the flavonoids 3-hydroxyflavone (flavonol), 5,7-dihydroxyflavone (chrysin), and 3',4' dihydroxyflavone in 50:50 (v/v) ethanol/water are determined by pH potentiometric and spectrophotometric titrations and the linear least-squares curve-fitting program Hyperquad. Over the entire range of pH and reagent concentrations spanning the titration experiments, the stoichiometry for iron-flavonoid complex formation was 1:1 for all three flavonoids examined. The three flavonoids were chosen for their hydroxy substitution pattern, with each possessing one of the three most commonly suggested sites for metal binding by the flavonoids. On the basis of the calculated stability constants, the intraflavonoid-binding site competition is illustrated as a function of pH via speciation curves. The curves indicate that the binding site comprised of the 3',4'-hydroxy substitutions, the catecholic site, is most influential for ferric complexation at the physiological pH of 7.4. The possibility for antioxidant activity by flavonoid chelation of ferric iron in the presence of other competitive physiological complexing agents is demonstrated through additional speciation calculations. PMID- 15826046 TI - Soybean oil based greases: influence of composition on thermo-oxidative and tribochemical behavior. AB - The biodegradable properties and lubricating ability of greases depend on both the base oil and the thickener. Mineral oils are the most widely used lubricant base fluids due to their inherent lubricity and low cost, but recent environmental awareness has forced consideration of the use of biodegradable fluids such as vegetable oils and certain synthetic fluids in grease formulations. This study presents data on the thermo-oxidation behavior and tribology of biodegradable greases formulated with soybean oil and different compositions of metal soap thickener. The composition of thickener has been varied by using fatty acids with different degrees of unsaturation and fatty acids of different chain lengths. The improvement of thermo-oxidation and tribological properties as a result of changing thickener/base oil ratio and the antioxidative effect of some suitable additives have also been evaluated. PMID- 15826047 TI - Detoxification of corn stover and corn starch pyrolysis liquors by ligninolytic enzymes of Phanerochaete chrysosporium. AB - Phanerochaete chrysosporium (ATCC 24725) shake flask culture with 3 mM veratryl alcohol addition on day 3 was able to grow and detoxify different concentrations of diluted corn stover (Dcs) and diluted corn starch (Dst) pyrolysis liquors [10, 25, and 50% (v/v)] in defined media. GC-MS analysis of reaction products showed a decrease and change in some compounds. In addition, the total phenolic assay with Dcs samples demonstrated a decrease in the phenolic compounds. A bioassay employing Lactobacillus casei growth and lactic acid production was developed to confirm the removal of toxic compounds from 10 and 25% (v/v) Dcs and Dst by the lignolytic enzymes, but not from 50% (v/v) Dcs and Dst. The removal did not occur when sodium azide or cycloheximide was added to Ph. chrysosporium culture media, confirming the participation of lignolytic enzymes in the detoxification process. A concentrated enzyme preparation decreased the phenolic compounds in 10% (v/v) corn stover and corn starch pyrolysis liquors to the same extent as the fungal cultures. PMID- 15826048 TI - Detoxification of corn stover and corn starch pyrolysis liquors by Pseudomonas putida and Streptomyces setonii suspended cells and plastic compost support biofilms. AB - Plant biomass can be liquefied into fermentable sugars (levoglucosan then to glucose) for the production of ethanol, lactic acid, enzymes, and more by a process called pyrolysis. During the process microbial inhibitors are also generated. Pseudomonas putida (ATCC 17484) and Streptomyces setonii75Vi2 (ATCC 39116) were employed to degrade microbial inhibitors in diluted corn stover (Dcs) and diluted corn starch (Dst) pyrolysis liquors. The detoxification process evaluation included measuring total phenols and changes in UV spectra, a GC-MS analysis, and a bioassay, which employed Lactobacillus casei subsp. rhamosus (ATCC 11443) growth as an indicator of detoxification. Suspended-cell cultures illustrated limited detoxification ability of Dcs and Dst. P. putida and S. setoniiplastic compost support (PCS) biofilm continuous-stirred-tank-reactor pure cultures detoxified 10 and 25% (v/v) Dcs and Dst, whereas PCS biofilm mixed culture also partially detoxified 50% (v/v) Dcs and Dst in repeated batch culture. Therefore, PCS biofilm mixed culture is the process of choice to detoxify diluted pyrolysis liquors. PMID- 15826049 TI - Thermal and high-pressure inactivation kinetics of polyphenol oxidase in Victoria grape must. AB - The inactivation kinetics of polyphenol oxidase (PPO) in freshly prepared grape must under high hydrostatic pressure (100-800 MPa) combined with moderate temperature (20-70 degrees C) was investigated. Atmospheric pressure conditions in a temperature range of 55-70 degrees C were also tested. Isothermal inactivation of PPO in grape must could be described by a biphasic model. The values of activation energy and activation volume of stable fraction were estimated as 53.34 kJ mol(-1) and -18.15 cm3 mol(-1) at a reference pressure of 600 MPa and reference temperature of 50 degrees C, respectively. Pressure and temperature were found to act synergistically, except in the high-temperature-low pressure region where an antagonistic effect was found. A third-degree polynomial model was successfully applied to describe the temperature/pressure dependence of the inactivation rate constants of the stable PPO fraction in grape must. PMID- 15826050 TI - Chemical indicators of heat treatment in fortified and special milks. AB - Carbohydrate and furosine contents in 12 commercial fortified and special milk samples (pasteurized goat's and ewe's milks; ultrahigh-temperature (UHT) goat's milk, UHT milks fortified with calcium, magnesium, fiber, or royal jelly and honey; and lactose-hydrolyzed milks) were analyzed. Except for lactose-hydrolyzed milks, furosine, lactose, lactulose, galactose, glucose, N-acetylgalactosamine, N acetylglucosamine, and myo-inositol contents were similar to the previously reported values for UHT or pasteurized milk samples. In lactose-hydrolyzed milks, lactulose was not detectable and lactose was present in low amount; high levels of glucose, galactose, fructose, tagatose, and furosine were also detected in this type of milk. Results found in commercial milks were compared to those obtained in laboratory-prepared UHT milks with lactose hydrolyzed prior to heating. Hydrolysis of lactose before thermal treatments promoted elevated accumulation of reducing sugars (galactose and glucose) that could be partially converted to the corresponding isomers (tagatose and fructose) during heating. In addition, the reducing sugars could also react with the amino groups of proteins, giving rise to the corresponding Amadori compound. According to the obtained results, heating prior to hydrolysis of lactose is suggested to avoid a considerable loss of available lysine. PMID- 15826051 TI - Release of nucleotides and nucleosides during yeast autolysis: kinetics and potential impact on flavor. AB - Nucleotides, particularly 5'-nucleotides, are important flavoring agents found in many foods and beverages. Their precise effect on the flavor of wines aged on lees has not been examined previously. In this study nucleotides and nucleosides released by yeast during autolysis in a model wine system and in Champagne wines were identified and quantified, and their impact on wine flavor was determined. Ribonucleotides only were detected in yeast autolysate and in Champagne wines. In wines ribonucleotides were quantified by tandem mass spectrometry coupled to HPLC. The maximum concentration of total nucleotides was very low with a maximum of approximately 3 mg/L in wine aged on yeasts for 9 years. In young wines the most important nucleotide was 5'-UMP, but after 2 years of aging its concentration decreased and the concentration of 5'-GMP slowly increased. The threshold values of the most representative nucleotides in Champagne wines were higher than the concentrations found in the same wines. However, it is known that there is synergism between the different nucleotides and also in the presence of glutamic acid. This phenomenon could explain the difference observed in descriptive profiles of wines spiked with nucleotides. PMID- 15826052 TI - Separation of membranes from acid-solubilized fish muscle proteins with the aid of calcium ions and organic acids. AB - Calcium chloride, and to a lesser extent MgCl2, aided in the separation of membranes by centrifugation from cod (Gadus morhua) muscle homogenates solubilized at pH 3 in the presence of citric acid or malic acid but not lactic acid. Adding citric acid and Ca2+ before solubilizing the cod muscle homogenates was needed for the effect. At 1 mM citric acid, 70-80% of the phospholipid and 25 30% of the protein were removed at 10 mM Ca2+. At 8 mM Ca2+, citric acid showed an optimal effect on phospholipid removal at 5 mM with 90% of the phospholipid and 35% of the protein removed. The treatment with citric acid and Ca2+ was also effective in separating the membrane from solubilized herring (Clupea harengus) muscle homogenate. Ca2+ and citric acid might exert their influence by disconnecting linkages between membranes and cytoskeletal proteins. PMID- 15826054 TI - Fate of pesticides during the winemaking process in relation to malolactic fermentation. AB - The effect of red wine malolactic fermentation on the fate of seven fungicides (carbendazim, chlorothalonil, fenarimol, metalaxyl, oxadixyl, procymidone, and triadimenol) and three insecticides (carbaryl, chlorpyrifos, and dicofol) was investigated. After malolactic fermentation using Oenococcus oeni, which simulated common Australian enological conditions, the concentrations of the active compounds chlorpyrifos and dicofol were the most significantly reduced, whereas the concentrations of chlorothalonil and procymidone diminished only slightly. The effect of these pesticides on the activity of the bacteria was also studied. Dicofol had a major inhibitory effect on the catabolism of malic acid, whereas chlorothalonil, chlorpyrifos, and fenarimol had only a minor effect. PMID- 15826053 TI - Polyphenol-rich phloem enhances the resistance of total serum lipids to oxidation in men. AB - In humans, polyphenol supplementation studies have resulted in inconsistent findings in lipid peroxidation. Our aim was to investigate the effects of a 4 week consumption of polyphenol-rich phloem on serum lipids and lipid peroxidation in the hydrophilic fraction of serum and on isolated lipoproteins. We conducted a randomized double-blind supplementation study consisting of 75 nonsmoking hypercholesterolemic men. Participants consumed 70 g daily of either rye bread (placebo) or phloem-fortified rye bread containing 31 mg (low polyphenol, LP) or 62 mg (high polyphenol, HP) of catechins. The ex vivo susceptibility of total serum lipids and VLDL and LDL to oxidation after copper induction was measured as a lag time to the maximal oxidation rate at the baseline and after the supplementation. In the HP group, an increase in the oxidation resistance of total serum lipids was observed (11.4%), while no effect was seen in the LP group (-0.8%) or in the placebo group (-1.0%) (p = 0.007). No differences were observed in the oxidation resistance of VLDL and LDL between the study groups. The phloem also increased in vitro oxidation resistance of serum lipids and radical scavenging activity (DPPH.) in a dose-dependent manner. Our results suggest that polyphenols may inhibit lipid peroxidation in the hydrophilic fraction of serum. PMID- 15826055 TI - Effect of different cooking methods on vegetable oxalate content. AB - Approximately 75% of all kidney stones are composed primarily of calcium oxalate, and hyperoxaluria is a primary risk factor for this disorder. Nine types of raw and cooked vegetables were analyzed for oxalate using an enzymatic method. There was a high proportion of water-soluble oxalate in most of the tested raw vegetables. Boiling markedly reduced soluble oxalate content by 30-87% and was more effective than steaming (5-53%) and baking (used only for potatoes, no oxalate loss). An assessment of the oxalate content of cooking water used for boiling and steaming revealed an approximately 100% recovery of oxalate losses. The losses of insoluble oxalate during cooking varied greatly, ranging from 0 to 74%. Because soluble sources of oxalate appear to be better absorbed than insoluble sources, employing cooking methods that significantly reduce soluble oxalate may be an effective strategy for decreasing oxaluria in individuals predisposed to the development of kidney stones. PMID- 15826056 TI - Kinetic analysis of nonisothermal differential scanning calorimetry of 1,3 dipalmitoyl-2-oleoylglycerol. AB - The crystallization of fats has been extensively studied because of its importance in the processing of food and food ingredients. Differential scanning calorimetry (DSC) is widely used in such studies. The aim of this study was to examine the determination of kinetic parameters from nonisothermal DSC crystallization of a model fat, 1,3-dipalmitoyl-2-oleoylglycerol. We applied peak and isoconversional methods to determine activation energies and compared these techniques with a nonparametric method, which separates the temperature dependence and degree of crystallization dependence of the crystallization rate. The Johnson-Mehl-Avrami-Erofeyev-Kolmogorov (JMAEK) model provided the best fit to the data, while the temperature dependence of the rate constant was best explained by a Vogel-Fulcher relationship, where the reference temperature was the melting point of the crystallizing species. PMID- 15826057 TI - Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola. AB - Since the 18th of April 2004, two new regulations, EC/1829/2003 on genetically modified food and feed products and EC/1830/2003 on traceability and labeling of GMOs, are in force in the EU. This new, comprehensive regulatory framework emphasizes the need of an adequate tracing system. Unique identifiers, such as the transgene genome junction region or a specific rearrangement within the transgene DNA, should form the basis of such a tracing system. In this study, we describe the development of event-specific tracing systems for transgenic maize lines Bt11, Bt176, and GA21 and for canola event GT73. Molecular characterization of the transgene loci enabled us to clone an event-specific sequence into a plasmid vector, to be used as a marker, and to develop line-specific primers. Primer specificity was tested through qualitative PCRs and dissociation curve analysis in SYBR Green I real-time PCRs. The primers were then combined with event-specific TaqMan probes in quantitative real-time PCRs. Calibration curves were set up both with genomic DNA samples and the newly synthesized plasmid DNA markers. It is shown that cloned plasmid GMO target sequences are perfectly suitable as unique identifiers and quantitative calibrators. Together with an event-specific primer pair and a highly specific TaqMan probe, the plasmid markers form crucial components of a unique and straighforward tracing system for Bt11, Bt176, and GA21 maize and GT73 canola events. PMID- 15826058 TI - Rapid tests to assess the antioxidant activity of Phaseolus vulgaris L. dry beans. AB - The antiradical activity of dry beans was evaluated in order to assess the validity of this test and to correlate the results with those achieved with the Folin-Ciocalteu method and with a rapid spectrophotometric method for the analysis of total flavonoids. Four landraces (12 samples) of common beans (Phaseolus vulgaris L.), collected in two regions of Italy (Tuscany and Basilicata) in three different years, were analyzed. The EC50 values ranged from 39 to 2810 mg sample/mg 1,1-diphenyl-2-picrylhydrazil radical. The phenolic content of each sample was expressed as gallic acid equivalents; it changed from 1.17 to 4.40 mg/g. The flavonoid content, expressed as mg of (+)-catechin per g of dry seeds, ranged from 0.24 to 1.43 mg/g. The qualiquantitative composition of polyphenols has been also elucidated by means of high-performance liquid chromatography (HPLC)-diode array detection and HPLC/MS. These investigations showed that rapid tests can contribute to assessing the quality of functional food. PMID- 15826059 TI - Effects of grinding and thermal treatments on hydrolysis susceptibility of pea proteins (Pisum sativum L.). AB - The effects of three particle sizes with two types of grindings and two thermal treatments on pea protein extraction (PE) and susceptibility to in vitro enzymatic hydrolysis (pepsin plus trypsin) were studied. Degrees of hydrolysis (DH) were calculated. Remaining peptides were detected by SDS-PAGE and identified by immunoblotting and MS/MS spectrometry. The increase in particle size decreased PE and DH due to a restricted access of solvents and enzymes to proteins. The thermal treatment induced a decrease in PE but did not modify DH. Heating improved legumin (alphaM) and convicilin pepsin hydrolyses but reduced the pea albumin 2 (PA2) hydrolysis. After pepsin and trypsin hydrolysis, only peptides from vicilin and lectin were identified by LC-MS/MS analyses, whatever the treatment. PMID- 15826060 TI - Cultivar and growing region determine the antioxidant polyphenolic concentration and composition of apples grown in New Zealand. AB - Evidence suggests that increasing consumption of fruit and vegetables contributes to improved health and well-being by providing protection from diseases including various cancers and cardiovascular disease. Although there is uncertainty about which components generate this effect, an attractive hypothesis is that the antioxidants are at least partly responsible. We measured the polyphenolic concentrations in 10 different apple cultivars grown commercially in New Zealand, each sourced from three different geographic regions. Our results showed that the concentration of polyphenolics varied among the apple cultivars, with Pacific Queen containing 2.7 times the amount of polyphenolics found in Cox's Orange. Furthermore, there were significant differences in polyphenolic concentrations in fruit from different regions for some cultivars but not for others. We also measured the polyphenolic concentrations in apple skin and flesh and found that on average 46% of the polyphenolics in whole apples were in the skin. Essentially all of the flavonols (quercetin derivatives) were present in the skin. To maximize the intake of apple polyphenols, it is necessary to consume apples of cultivars with high polyphenolic concentrations such as Pacific Queen and include the skin. Our results also showed that there is potential for promoting apple fruit from specific geographical regions because they contained elevated concentrations of antioxidant polyphenolic compounds. PMID- 15826061 TI - Typification of cider brandy on the basis of cider used in its manufacture. AB - A study of typification of cider brandies on the basis of the origin of the raw material used in their manufacture was conducted using chemometric techniques (principal component analysis, linear discriminant analysis, and Bayesian analysis) together with their composition in volatile compounds, as analyzed by gas chromatography with flame ionization to detect the major volatiles and by mass spectrometric to detect the minor ones. Significant principal components computed by a double cross-validation procedure allowed the structure of the database to be visualized as a function of the raw material, that is, cider made from fresh apple juice versus cider made from apple juice concentrate. Feasible and robust discriminant rules were computed and validated by a cross-validation procedure that allowed the authors to classify fresh and concentrate cider brandies, obtaining classification hits of >92%. The most discriminating variables for typifying cider brandies according to their raw material were 1 butanol and ethyl hexanoate. PMID- 15826062 TI - Comparison of the chemical composition of extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extraction versus standard hot water or 70% ethanol extraction. AB - The aqueous extract of American skullcap (Scutellaria lateriflora L. (S. lateriflora), Lamiaceae) has been traditionally used by North American Indians as a nerve tonic and for its sedative and diuretic properties. Recent reports stated that flavonoids and possibly amino acids are responsible for the anxiolytic activity. As a part of our search for environmentally friendly solvents to extract the active components from medicinal plants, we used S. lateriflora in a comparison of accelerated solvent extraction (ASE) using water, and supercritical fluid extraction (SFE) using CO2 and 10% EtOH as modifier, at different temperatures. Flavonoids and amino acids were quantified by HPLC-UV and HPLC-MS, respectively. The flavonoid content was compared with conventional extraction methods (hot water extraction and 70% ethanol). The use of ASE at 85 degrees C with water as solvent gave the best results for flavonoid glycosides and amino acids, whereas SFE gave higher yields of flavonoid aglycones. However, the results obtained for total flavonoids were not significatively superior to hot water extraction or 70% aqueous EtOH extract. PMID- 15826063 TI - Pressure and temperature stability of 5-methyltetrahydrofolic acid: a kinetic study. AB - Detailed kinetic studies of [6S] and [6RS] 5-methyltetrahydrofolic acid (5-CH3 H4folate) degradation during thermal (from 60 to 90 degrees C) and high pressure/thermal (from 30 to 45 degrees C; from 200 to 700 MPa) treatments were carried out. The results confirmed that the temperature and pressure induced degradation kinetics of [6S] 5-CH3-H4folate were identical (within 95% confidence interval) with those of [6RS] 5-CH3-H4folate. Under equal processing conditions, the estimated degradation rate constants (k), activation energy (E(a)), and activation volume (V(a)) values of [6S] and [6RS] 5-CH3-H4folate were the same (95% confidence interval). The modified thermodynamic model proposed by Nguyen and co-workers (J. Agric. Food Chem. 2003, 51, 3352-3357) to describe the pressure and temperature dependence of the rate constant for folate degradation was reevaluated. PMID- 15826064 TI - Nutritional potential of raw and free alpha-galactosides lupin (Lupinus albus Var. multolupa) seed flours. Effect of phytase treatment on nitrogen and mineral dialyzability. AB - The effect of the removal of alpha-galactosides from Lupinus albus L. var. multolupa on the chemical composition of the prepared flour and the dialyzability of N, total P, Ca, Mg, Fe, Zn, and Mn was studied. The extraction process caused a significant increase in total and insoluble nitrogen contents and decreased the amount of soluble protein nitrogen. However, neither these changes nor treatment with phytase seemed to considerably affect in vitro protein digestibility. Except for Ca and Cu, total mineral contents were significantly reduced by the extraction process. The process also caused a significant reduction in the dialyzability of all the minerals studied except P. The decrease in mineral dialyzability was partially counteracted by phytase treatment in amounts of 250 500 phytase units/kg of lupin flour. In the case of Fe, mineral dialyzability did not differ significantly between the two lupin flours studied with treatment with 500 phytase units/kg. Zinc dialyzability was the most efficiently improved by phytase treatment (P < 0.0001), followed by P, Fe, and Mn, and finally by Ca and Mg (P < 0.05). PMID- 15826065 TI - The use of carbohydrate profiles and chemometrics in the characterization of natural honeys of identical geographical origin. AB - A study of the real possibilities of carbohydrate profiles and chemometrics to characterize the botanical origin of honey from a single geographical area, the Province of Soria (Spain), is presented. To this end, 14 carbohydrates were quantified using high-performance liquid chromatography (HPLC) with pulsed amperometric detection (PAD) in 77 natural honeys, the botanical origins of which were ling, spike lavender, French lavender, thyme, forest, and multifloral. Principal component analysis has been employed as a first approach to characterize the honey samples analyzed, showing similarities between spike lavender and multifloral honeys. The best discrimination among groups is obtained when four canonical discriminant analyses were carried out sequentially, origin by origin, achieving an overall percentage of success of 90% following cross validation. PMID- 15826066 TI - Identification and characterization of anthocyanins by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry in common foods in the United States: vegetables, nuts, and grains. AB - Anthocyanins in common foods in the United States, other than fruits and berries, were identified and characterized by high-performance liquid chromatography (HPLC)-electrospray ionization-tandem mass spectrometry coupled with diode array detection. Of all of the 40+ vegetables, nuts, and grains screened, seven vegetables, one nut, and one grain were found to contain anthocyanins; the number of anthocyanins detected varied from two in pistachio nuts to 34 in red radishes. The individual anthocyanins were identified by comparing their mass spectrometric data and retention times with those of standards, published data, and reference food samples. In all of the samples analyzed, except for sorghum, only six common anthocyanidins (delphinidin, cyanidin, pelargonidin, petunidin, peonidin, and malvidin) were found as their glycosides. Anthocyanins in certain vegetables such as red cabbage and red radish were highly conjugated with sugars and acylated groups, and thus, their structures were very complicated. Eight different either aliphatic or aromatic acylated groups (acetoyl, coumaroyl, malonoyl, p hydroxybenzoyl, feruoyl, caffeoyl, sinapoyl, and oxaloyl) were identified in the anthocyanins. In addition to glucose, six other sugar moieties (galactose, xylose, rhamnose, rutinose, sambubiose, and laminaribiose) were observed. Three varieties of sorghum were found to contain 3-deoxyanthocyanidins and their derivatives as major anthocyanins. A number of new anthocyanins were identified in the foods studied. This paper presents complete HPLC profiles and MS spectrometric data, obtained under the same experimental conditions, for common vegetables, pistachio nuts, and sorghum that contain anthocyanins. PMID- 15826067 TI - Seasonal variations in the level of plant constituents in greenhouse production of cherry tomatoes. AB - The content of selected plant constituents was measured in cherry tomatoes (Lycopersicon esculentumMill. cv. Jennita) during conventional Norwegian tomato production in a greenhouse from May until October 2004. Samples were collected according to standard production procedure with orange-yellow colored fruits at weight in the range of 12.4-19.3 g and size in the range of 28.9-33.0 mm (diameter). The content of selected compounds based on 100 g FW were found to vary in the following range during the season: 7.38-28.38 mg of chalconaringenin, 0.32-0.92 mg of rutin, 0.24-1.06 mg of chlorogenic acid, 5.60-20.02 mg of ascorbic acid, 1.60-5.54 mg of lycopene, and 0.37-0.55 mg beta-carotene. Only minute amounts of naringenin together with kaempferol 3-rutinoside and caffeic acid, which previously have been reported from tomatoes, were detected. The content of chalconaringenin to rutin and that of lycopene to beta-carotene showed a strong correlation during the season (p < 0.001). The content of total phenolics and methanol-soluble antioxidants also showed a correlation (p < 0.001), and were found in the range 14.6-32.6 mg of gallic acid equivalents (GAE)/100 g fresh weight (FW) and 445-737 micromol of Fe(II)/100 g FW, respectively. Seasonal variation in the level of plant constituents is shown to be related to photon flux density and fertilization level. PMID- 15826068 TI - Syntheses, antifeedant activity, and QSAR analysis of new oxa(thia)diazolyl 3(2H) pyridazinones. AB - Twenty-eight new oxa(thia)diazolyl 3(2H)-pyridazinone derivatives were synthesized. Some compounds showed good chronic growth activities against the armyworm, Pseudaletia separata (Walker). Their EC50 values were determined in vivo. Nineteen 2-tert-butyl-4-chloro-5-[5'-aryl-2'-(1',3',4'-oxadiazolyl)methoxy] 3(2H)-pyridazinones were quantitatively analyzed using the QSAR procedure. The partial least squares method was applied to find relationships between activity and the physicochemical data. The results showed that dipole moment, molar refractivity, and log P are identified as critical parameters for chronic growth effects. PMID- 15826069 TI - Characterization of aroma compounds responsible for the rosy/floral flavor in Cheddar cheese. AB - The aroma-active compounds that contribute to the rosy/floral flavor in Cheddar cheese were characterized using both instrumental and sensory techniques. Two cheeses (>12 months old) with rosy/floral flavor and two Cheddar cheeses of similar ages without rosy/floral flavors were selected. After direct solvent extraction/solvent-assisted flavor evaporation and separation into neutral/basic and acidic fractions, samples were analyzed by gas chromatography-olfactometry with aroma extract dilution analysis. Selected compounds were quantified using internal standard methodology. Some of the intense aroma-active compounds in the neutral basic fraction of the rosy/floral cheeses included 2-phenethanol (rosy), phenylethyl acetate (rosy), and phenylacetaldehyde (rosy/floral). Quantification, threshold analysis, and sensory analysis of model cheeses confirmed that increased concentrations of phenylacetaldehyde and phenylacetic acid caused rosy/floral flavor when spiked into Cheddar cheese. PMID- 15826070 TI - Relationship of ethylene biosynthesis to volatile production, related enzymes, and precursor availability in apple peel and flesh tissues. AB - Regulation of ethylene biosynthesis or action has a major effect on volatiles production in apples. To understand the biochemical processes involved, we used Greensleeves apples from a transgenic line with a high suppression of ethylene biosynthesis. The study was focused at the level of the aroma volatile-related enzymes, including alcohol acyltransferase (AAT), alcohol dehydrogenase (ADH), and lipoxygenase (LOX) and at the level of amino acids and fatty acids as aroma volatile precursors in peel and flesh tissues. In general, volatile production, enzyme activity levels, and precursor availability were higher in the peel than the flesh and were differentially affected by ethylene regulation. AAT enzyme activity showed a clear pattern concomitant with ethylene regulation. Contrarily, ADH and LOX seem to be independent of ethylene modulation. Isoleucine, an important precursor of aroma compounds including 2-methylbutanoate esters, showed a major increase in the peel during ripening and responded significantly to ethylene regulation. Other important aroma volatiles precursors, like linoleic and linolenic acid, showed an accumulation during ripening associated with increases in aldehydes. The significance of these changes in relation to aroma volatile production is discussed. PMID- 15826071 TI - Carotenoid pigmentation affects the volatile composition of tomato and watermelon fruits, as revealed by comparative genetic analyses. AB - Tomato near-isogenic lines differing in fruit carotenogenesis genes accumulated different aroma volatiles, in a strikingly similar fashion as compared to watermelon cultivars differing in fruit color. The major volatile norisoprenoids present in lycopene-containing tomatoes and watermelons were noncyclic, such as geranial, neral, 6-methyl-5-hepten-2-one, 2,6-dimethylhept-5-1-al, 2,3 epoxygeranial, (E,E)-pseudoionone, geranyl acetone, and farnesyl acetone, seemingly derived from lycopene and other noncyclic tetraterpenoids. Beta-ionone, dihydroactinodiolide, and beta-cyclocitral were prominent in both tomato and watermelon fruits containing beta-carotene. Alpha-ionone was detected only in an orange-fleshed tomato mutant that accumulates delta-carotene. A yellow flesh (r) mutant tomato bearing a nonfunctional psy1 gene and the yellow-fleshed watermelon Early Moonbeam, almost devoid of carotenoid fruit pigments, also lacked norisoprenoid derivatives and geranial. This study provides evidence, based on comparative genetics, that carotenoid pigmentation patterns have profound effects on the norisoprene and monoterpene aroma volatile compositions of tomato and watermelon and that in these fruits geranial (trans-citral) is apparently derived from lycopene in vivo. PMID- 15826072 TI - Development and validation of a dynamic vapor sorption-fast gas chromatography flame ionization detection method for rapid analysis of volatile release from glassy matrices. AB - Historically, saturated salt solutions in desiccators have been used to investigate the effects of increasing relative humidity and temperature on the volatile retention efficiency of amorphous glasses. Obtaining data using desiccators is a static process that gives the researcher discrete data points with which to draw conclusions. Dynamic vapor sorption (DVS; SMS, London, U.K.) is a humidification system that creates specific relative humidity and temperature environments within a chamber that contains the material being investigated. This study had two specific aims: (1) to develop a DVS-fast GC-FID method that dynamically evaluates the effects of humidification and temperature increases on volatile release from amorphous carbohydrate glasses and (2) to evaluate the validity of the DVS-fast GC-FID method. Artificial cherry Durarome (Firmenich, Plainsboro, NJ) was used as the model system. The DVS-fast GC-FID method proved to be an innovative, accurate, and precise technique that can be used to conduct humidification/temperature-volatile release studies. PMID- 15826073 TI - Antioxidant activity in lingonberries (Vaccinium vitis-idaea L.) and its inhibitory effect on activator protein-1, nuclear factor-kappaB, and mitogen activated protein kinases activation. AB - Lingonberry has been shown to contain high antioxidant activity. Fruits from different cultivars of lingonberry (Vaccinium vitis-idaea L.) were evaluated for fruit quality, antioxidant activity, and anthocyanin and phenolic contents. The fruit soluble solids, titratable acids, antioxidant capacity, and anthocyanin and phenolic contents varied with cultivars. Lingonberries contain potent free radical scavenging activities for DPPH*, ROO*, *OH, and O2*- radicals. Pretreatment of JB6 P+ mouse epidermal cells with lingonberry extracts produced a dose-dependent inhibition on the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12-O-tetradecanoylphorbol-13 acetate (TPA) or ultraviolet-B (UVB). Lingonberry extract blocked UVB-induced phosphorylation of the mitogen-activated protein kinase (MAPK) signaling members ERK1, ERK2, p38, and MEK1/2 but not JNK. Lingonberry extract also prevented TPA induced phosphorylation of ERK1, ERK2, and MEK1/2. Results of soft agar assays indicated that lingonberry extract suppressed TPA-induced neoplastic transformation of JB6 P(+) cells in a dose-dependent manner. Lingonberry extract also induced the apoptosis of human leukemia HL-60 cells in a dose-independent manner. These results suggest that ERK1, ERK2, and MEK1/2 may be the primary targets of lingonberry that result in suppression of AP-1, NF-kappaB, and neoplastic transformation in JB6 P(+) cells and causes cancer cell death by an apoptotic mechanism in human leukemia HL-60 cells. PMID- 15826074 TI - Inhibitory effect of naturally occurring flavonoids on the formation of advanced glycation endproducts. AB - The objective of this study was to investigate the inhibitory effect of naturally occurring flavonoids on individual stage of protein glycation in vitro using the model systems of delta-Gluconolactone assay (early stage), BSA-methylglyoxal assay (middle stage), BSA-glucose assay, and G.K. peptide-ribose assay (last stage). In the early stage of protein glycation, luteolin, qucertin, and rutin exhibited significant inhibitory activity on HbA1C formation (p < 0.01), which were more effective than that of aminoguanidine (AG, 10 mM), a well-known inhibitor for advanced glycation endproducts (AGEs). For the middle stage, luteolin and rutin developed more significant inhibitory effect on methylglyoxal medicated protein modification, and the IC50's were 66.1 and 71.8 microM, respectively. In the last stage of glycation, luteolin was found to be potent inhibitors of both the AGEs formation and the subsequent cross-linking of proteins. In addition, phenyl-tert-butyl-nitron served as a spin-trapping agent, and electron spin resonance (ESR) was used to explore the possible mechanism of the inhibitory effect of flavonoids on glycation. The results indicated that protein glycation was accompanied by oxidative reactions, as the ESR spectra showed a clear-cut radical signal. Statistical analysis showed that inhibitory capability of flavonoids against protein glycation was remarkably related to the scavenging free radicals derived from glycoxidation process (r = 0.79, p < 0.01). Consequently, the inhibitory mechanism of flavonoids against glycation was, at least partly, due to their antioxidant properties. PMID- 15826075 TI - Changes in carotenoid and ascorbic acid contents in fruits of different tomato genotypes related to the depletion of UV-B radiation. AB - The aim of the present study was to investigate if the depletion of UV-B radiation affected the most representative carotenoids as well as the ascorbic acid content in tomato fruits, harvested at both breaker and firm red stages. To do this, three tomato genotypes, DRW 5981, HP 1, and Esperanza, were grown inside a greenhouse either covered with polyethylene transparent to UV-B or depleted of UV-B by a special covering film. The antioxidant properties of the fruits were evaluated on the water-insoluble fractions according to the ABTS method. UV-B effect on antioxidant activity was negligible in DRW and HP 1 genotypes, whereas it was detrimental in Esperanza at both ripening stages. This genotype seems to have a negligible capability of accumulating carotenoids and a great susceptibility to detrimental effects of UV-B; conversely, the DRW genotype shows high carotenoid levels under sunlight conditions and a further promotion by UV-B. On the other hand, the HP 1 mutant displays an intermediate behavior and represents the only genotype favored by UV-B with respect to ascorbic acid accumulation. PMID- 15826076 TI - Denaturation and aggregation of three alpha-lactalbumin preparations at neutral pH. AB - The denaturation and aggregation of reagent-grade (Sigmaalpha-La), ion-exchange chromatography purified (IEXalpha-La), and a commercial-grade (Calpha-La) alpha lactalbumin were studied with differential scanning calorimetry (DSC), polyacrylamide gel electrophoresis, and turbidity measurement. All three preparations had similar thermal denaturation temperatures with an average of 63.7 degrees C. Heating pure preparations of alpha-lactalbumin produced three non native monomer species and three distinct dimer species. This phenomenon was not observed in Calpha-La. Turbidity development at 95 degrees C (tau95 degrees C) indicated that pure preparations rapidly aggregate at pH 7.0, and evidence suggests that hydrophobic interactions drove this phenomenon. The Calpha-La required 4 times the phosphate or excess Ca2+ concentrations to develop a similar tau95 degrees C to the pure preparations and displayed a complex pH-dependent tau95 degrees C behavior. Turbidity development dramatically decreased when the heating temperature was below 95 degrees C. A mechanism is provided, and the interrelationship between specific electrostatic interactions and hydrophobic attraction, in relation to the formation of disulfide-bonded products, is discussed. PMID- 15826077 TI - Conformational stability of the potato serine protease inhibitor group. AB - The thermal unfolding of potato serine protease inhibitor (PSPI), the most abundant protease inhibitor group in potato tuber, was measured using far UV CD spectroscopy, fluorescence spectroscopy, and DSC. The results indicate that the thermal as well as the guanidinium-induced unfolding of PSPI occurs via a non-two state mechanism in which at least one stable intermediate is present. Additionally, the occurrence of aggregation, especially at low scan rates, increases the apparent cooperativity of the unfolding and makes the system kinetically rather than thermodynamically controlled. Aggregate formation seems to occur via a specific mechanism of which PSPI in a tetrameric form is the end product and which may involve disulfide interchanges. PMID- 15826078 TI - Influence of binding of sodium dodecyl sulfate, all-trans-retinol, palmitate, and 8-anilino-1-naphthalenesulfonate on the heat-induced unfolding and aggregation of beta-lactoglobulin B. AB - Heat treatment of bovine beta-lactoglobulin B (beta-LG) causes it to partially unfold and aggregate via hydrophobic association and intra- and interprotein disulfide bonds. The first stage, which involves a "loosening" of the native structure, is influenced by the environmental conditions, such as pressure, pH, and added solutes. In the present study, four potential beta-LG ligands [palmitate, sodium dodecyl sulfate (SDS), 8-anilino-1-naphthalenesulfonate (ANS), and all-trans-retinol (retinol)] were added to beta-LG solutions prior to heat treatment for 12 min at temperatures between 40 and 93 degrees C. The extent of the changes in secondary and tertiary structures, unfolding, and aggregation at 20 degrees C were determined by circular dichroism, fluorescence, and alkaline- and SDS-polyacrylamide gel electrophoresis (PAGE). Both palmitate and SDS stabilized the native structure of beta-LG against heat-induced structural flexibility, subsequent unfolding, and denaturation. Retinol was less effective, probably because of its lower affinity for the calyx-binding site, and ANS did not stabilize beta-LG, suggesting that ANS did not bind strongly in the calyx. It was also noted that holding a beta-LG solution with added SDS or ANS promoted the formation of a hydrophobically associated non-native dimer. PMID- 15826079 TI - Purification and partial characterization of broccoli (Brassica oleracea Var. Italica) peroxidases. AB - Three peroxidase (POD) isoenzymes were purified from a soluble extract of broccoli stems. The acidic and neutral PODs were purified to homogeneity by using ion exchange and hydrophobic chromatography. The basic POD was purified by cation exchange and gel filtration chromatography. The neutral and basic PODs had molecular masses of approximately 43 kDa, and the acidic POD had a molecular mass of 48 kDa by SDS-PAGE. pI was approximately 4, 5, and 8 for acidic, neutral, and basic PODs, respectively. Optimum activity using guaiacol as the H donor was obtained at pH approximately 6 for both neutral and basic PODs and at pH approximately 4 for acidic POD. All three of the purified isoenzymes are glycosylated. Reaction rates with various substrates including guaiacol, guaiacol/MBTH, DMAB/MBTH, and ferulic acid/MBTH were different among the isoenzymes. K(m) and amino acid composition were also determined. PMID- 15826080 TI - Heat inactivation and reactivation of broccoli peroxidase. AB - Heat inactivation characteristics differed for acidic (A), neutral (N), and basic (B) broccoli peroxidase. At 65 degrees C, A was the most heat stable followed by N and B. The activation energies for denaturation were 388, 189, and 269 kJ/mol for A, N, and B, respectively. Reactivation of N occurred rapidly, within 10 min after the heated enzyme was cooled and incubated at room temperature. The extent of reactivation varied from 0 to 50% depending on the isoenzyme and heating conditions (temperature and time). The denaturation temperature allowing the maximum reactivation was 90 degrees C for A and horseradish peroxidase (HRP) and 70 and 80 degrees C for B and N, respectively. In all cases, heat treatment at low temperatures for long times prevented reactivation of the heated enzymes. Calcium (5 mM) increased the thermal stability of N and B but had no effect on reactivation. The presence of 0.05% bovine serum albumin decreased thermal stability but increased the extent of reactivation of A.. PMID- 15826081 TI - Changes in plasma lipid and antioxidant activity in rats as a result of naringin and red grapefruit supplementation. AB - The aim of this investigation was to compare the influence of naringin versus red grapefruit juice on plasma lipid levels and plasma antioxidant activity in rats fed cholesterol-containing and cholesterol-free diets. The antioxidant activity of a correlated quantity of red grapefruit juice was higher than that of naringin. Forty-two male Wistar rats were randomly divided into six groups of 7 named control, naringin, grapefruit, Chol, Chol/naringin, and Chol/grapefruit. The rats of the control group were fed basal diet (BD) and 1-2 mL of distilled water. To the BD of the other five groups were added 0.46-0.92 mg of naringin dissolved in 1-2 mL of distilled water (naringin), 1-2 mL of red grapefruit juice (grapefruit), 1% of nonoxidized cholesterol (NOC) and 1-2 mL of distilled water (Chol), 1% of NOC and 0.46-0.92 mg of naringin in 1-2 mL of water (Chol/naringin), and 1% of NOC and 1-2 mL of red grapefruit juice (Chol/grapefruit). After 30 days of different feeding, it was found that diets supplemented with red grapefruit juice and to a lesser degree with naringin improved the plasma lipid levels mainly in rats fed cholesterol and increased the plasma antioxidant activity. In conclusion, naringin is a powerful plasma lipid lowering and plasma antioxidant activity increasing flavonone. However, fresh red grapefruit is preferable than naringin: it more effectively influences plasma lipid levels and plasma antioxidant activity and, therefore, could be used as a valuable supplement for disease-preventing diets. PMID- 15826082 TI - Origin of the high-frequency resonances in 1H NMR spectra of muscle tissue: an in vitro slow magic-angle spinning study. AB - High-resolution slow magic-angle spinning (150 Hz) 1H PASS NMR spectroscopy is performed on intact excised rat m. tibialis anterior. Untreated muscles and muscles in vitro incubated in Krebs-Ringers buffer based on deuterium oxide are investigated. In the high-frequency region of the 1H NMR spectra, resonances from H4 (approximately 7.1-7.2 ppm) and H2 (approximately 8.2-8.5 ppm) in histidine are observed. In addition, a resonance appears at 6.7 ppm for the untreated muscles. However, this resonance is absent in muscles following incubation in deuterium oxide. On the basis of its behavior in deuterium oxide combined with supplementary measurements for creatine solutions, the 6.7 ppm resonance is ascribed to the amino protons in creatine. Moreover, the present study demonstrates that the observation of the 6.7 ppm resonance depends on pH, which explains earlier reports stating its occasional appearance. Finally, measurements on solutions of ATP/AMP and histidine indicate that both ATP/AMP and histidine contribute to the resonances at approximately 8.2-8.5 ppm in the 1H NMR spectra of muscle tissue. PMID- 15826083 TI - Role of beta-adrenoceptor signaling and AMP-activated protein kinase in glycolysis of postmortem skeletal muscle. AB - Postmortem glycolysis is directly linked to the incidences of PSE (pale, soft, and exudative) and DFD (dark, firm, and dry) meats, which cause significant economic loss to the meat industry. However, mechanisms controlling postmortem glycolysis are unclear. The objective of this study was to determine the role of beta-adrenoceptor signaling and AMP-activated protein kinase (AMPK) in postmortem glycolysis. Eighteen 2 month old C57BL/6J female mice were randomly separated into three groups. Group I received an intraperitoneal injection of saline solution only and served as the control; group II received a saline injection and then were forced to swim for 1 min; and group III received an injection of propranolol (1 mg/kg) in saline. In addition, six C57BL/6J female AMPK knockout mice were assigned to group IV, which received a saline injection and were forced to swim for 1 min. The longissimus dorsi muscle was sampled at 0, 1, and 24 h postmortem for pH and enzyme activity measurements. The objective is to elucidate the roles of beta-adrenoceptor signaling and AMPK in the glycolysis of postmortem muscle. Results showed that AMPK activity had a major role in determining the ultimate muscle pH, with an ultimate pH for control mice of 6.16 and AMPK knockout mice of 6.48. The beta-adrenoceptor signaling is essential for initial rapid glycolysis. Blocking beta-adrenoceptor signaling prevented the initial pH decline induced by stress. Activation of beta-adrenoceptor signaling due to preslaughter stress activates glycogen phosphorylase, resulting in a rapid glycolysis shortly after slaughter. On the other hand, the activation of AMPK is important for maintaining the activity of glycogen phosphorylase and pyruvate kinase, leading to a sustained glycolysis and a low ultimate pH. PMID- 15826084 TI - Iron and zinc in vitro availability in pearl millet flours (Pennisetum glaucum) with varying phytate, tannin, and fiber contents. AB - Simulations of gastro-intestinal digestion, used to estimate in vitro iron and zinc availability, were performed on two kinds of samples: (i) samples with decreased phytate contents from whole pearl millet flour and (ii) nondephytinized or dephytinized samples from two pearl millet grain fractions, a decorticated fraction with low fiber and tannin contents and a bran fraction with high fiber and tannin contents. Iron and zinc in vitro availabilities of whole pearl millet flour were significantly improved by phytate degradation, even if the IP6 were not all degraded. Total dephytinization of decorticated fraction led to a marked increase in iron and zinc in vitro availabilities, but that of bran fraction had no effect on either iron or zinc in vitro availability. Even if phytates are involved in reducing in vitro iron and zinc availability in pearl millet flour, fibers and tannins play an important role by chelating a high proportion of iron and zinc in grain hulls. PMID- 15826085 TI - Quantitation of carcinogenic heterocyclic aromatic amines and detection of novel heterocyclic aromatic amines in cooked meats and grill scrapings by HPLC/ESI-MS. AB - A tandem solid-phase extraction method was used to isolate carcinogenic heterocyclic aromatic amines (HAAs) from cooked meats. The following 10 HAAs were identified by HPLC/ESI-MS/MS: 2-amino-9H-pyrido[2,3-b]indole (2-AalphaC), 2-amino 3-methyl-9H-pyrido[2,3-b]indole (MeAalphaC), 2-amino-3-methylimidazo[4,5 f]quinoline (IQ), 2-amino-3-methylimidazo[4,5-f]quinoxaline (IQx), 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline (8-MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5 f]quinoxaline (4,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino 1,7,9-trimethylimidazo[4,5-g]quinoxaline (7,9-DiMeIgQx), and 2-amino-1 methylimidazo[4,5-b]quinoline (IQ[4,5-b]); the latter HAA has not previously been reported in cooked meats. The concentrations of these HAAs ranged from <0.03 to 15 ppb in cooked meats and poultry, to 75 ppb in cooked beef extract, and to 85 ppb in grill scrapings. The product ion scan mode was used to confirm the identities of these HAAs. Six other compounds were detected that appear to contain the N-methylimidazoquinoxaline skeleton on the basis of their product ion spectra, and these compounds are probable isomers of IQx, 8-MeIQx, and DiMeIQx. A number of known HAAs and novel HAAs of unknown genotoxic potential are formed at appreciable levels in cooked meats. PMID- 15826086 TI - Addition of glycine reduces the content of acrylamide in cereal and potato products. AB - The effects of adding amino acids on the content of acrylamide in potato crisps, French fries, flat breads, and bread crusts were investigated. Addition of glycine or glutamine during blanching of crisps reduced the amount of acrylamide by approximately 30% compared to no addition. No effect was found in French fries. Addition of glycine during doughmaking significantly reduced acrylamide in both flat breads and bread crusts. In bread crusts the reduction of acrylamide ranged from 50 to >90% depending on the baking condition. In flat breads the reduction varied between 60 and >95%. PMID- 15826087 TI - Correlation between surface photovoltage and blend morphology in polyfluorene based photodiodes. AB - We present a microscopic study of photoinduced charge generation in polyfluorene based photovoltaic structures. The sub-100 nm lateral resolution of scanning Kelvin probe microscopy allows characterizing the three-dimensional structure of thin films of blends of poly-(9,9'-dioctylfluorene-co-benzothiadiazole) (F8BT) and poly-(9,9'-dioctylfluorene-co-bis-N,N'-(4-butylphenyl)-bis-N,N'-phenyl-1,4 phenylenediamine) (PFB). From the strong correlation between surface photovoltage and blend morphology, we propose a simple model for the lateral and vertical film structure identifying in particular those regions with the most efficient conduction pathway for the photocurrent. PMID- 15826088 TI - Shell distribution on colloidal CdSe/ZnS quantum dots. AB - Scanning transmission electron microscopy (STEM) coupled with electron energy loss spectroscopy (EELS) was used to determine the chemical distribution of semiconductor shell material around colloidal core-shell CdSe/ZnS quantum dots (QDs). EELS signals from positions around the QD indicate a well-defined shell of ZnS surrounding the CdSe core, but the distribution of the shell material is highly anisotropic. This nonuniformity may reflect the differences in chemical activity of the crystal faces of the core QD and implies a nonoptimal QD surface passivation. PMID- 15826089 TI - Electrical conductivity of ferritin proteins by conductive AFM. AB - Electrical conductivity measurements were performed on single apoferritin and holoferritin molecules by conductive atomic force microscopy. Conductivity of self-assembled monolayer films of ferritin molecules on gold surfaces was also measured. Holoferritin was 5-15 times more conductive than apoferritin, indicating that for holoferritin most electron-transfer goes through the ferrihydrite core. With 1 V applied, the average electrical currents through single holoferritin and apoferritin molecules were 2.6 pA and 0.19 pA, respectively. PMID- 15826090 TI - Nanoscale morphology of high-performance polymer solar cells. AB - Transmission electron microscopy and electron diffraction are used to study the changes in morphology of composite films of poly(3-hexylthiophene) (P3HT) and a methanofullerene derivative (PCBM) in bulk heterojunction solar cells. Thermal annealing produces and stabilizes a nanoscale interpenetrating network with crystalline order for both components. P3HT forms long, thin conducting nanowires in a rather homogeneous, nanocrystalline PCBM film. Both the improved crystalline nature of films and increased but controlled demixing between the two constitutes therein after annealing explains the considerable increase of the power conversion efficiency observed in these devices. PMID- 15826091 TI - Gold nanoparticles quench fluorescence by phase induced radiative rate suppression. AB - The fluorescence quantum yield of Cy5 molecules attached to gold nanoparticles via ssDNA spacers is measured for Cy5-nanoparticle distances between 2 and 16 nm. Different numbers of ssDNA per nanoparticle allow to fine-tune the distance. The change of the radiative and nonradiative molecular decay rates with distance is determined using time-resolved photoluminescence spectroscopy. Remarkably, the distance dependent quantum efficiency is almost exclusively governed by the radiative rate. PMID- 15826092 TI - Progression of respiratory syncytial virus infection monitored by fluorescent quantum dot probes. AB - We report the use of quantum dots (QDs) to identify the presence and monitor the progression of respiratory syncytial virus (RSV) infection over time by labeling the F and G proteins. In addition, co-localization of these viral proteins was shown using confocal microscopy. The implications of these results are that QDs may provide a method for early, rapid detection of viral infection and open the door for future studies of the intricate spatial features cell trafficking of viral proteins. PMID- 15826093 TI - A virus-based nanoblock with tunable electrostatic properties. AB - Five different "HIS tag" mutants of cowpea mosaic virus were made by genetically introducing six contiguous histidine residues at various locations on the virus capsid. The mutant particles showed differential affinity for binding nickel, and their electrostatic properties could be controlled as a function of the protonation state of the exposed histidine sequence. The specific addressability of the HIS tag was corroborated by the selective modification of the histidine sequence with nanogold cross-linked to the Ni-NTA moiety. PMID- 15826094 TI - Peptide-conjugated quantum dots activate neuronal receptors and initiate downstream signaling of neurite growth. AB - Quantum dots (QDs) could serve as fluorescent scaffolds for effecting specific physiological and pharmacological responses in cells. Here, we conjugate the peptide ligand betaNGF to QD surfaces, and confirm surface modification and single QD nanostructure using AFM. We show that betaNGF-QDs retain bioactivity, activate TrkA receptors, and initiate neuronal differentiation in PC12 cells. Receptor-evoked activity of QD-immobilized ligands has wide-ranging implications for the development of molecular tools and therapeutics targeted at understanding and regulating cell function. PMID- 15826095 TI - Electrochemical nanofabrication using crystalline protein masks. AB - We have developed a simple and robust method to fabricate nanoarrays of metals and metal oxides over macroscopic substrates using the crystalline surface layer (S-layer) protein of Deinococcus radiodurans as an electrodeposition mask. Substrates are coated by adsorption of the S-layer from a detergent-stabilized aqueous protein extract, producing insulating masks with 2-3 nm diameter solvent accessible openings to the deposition substrate. The coating process can be controlled to achieve complete or fractional surface coverage. We demonstrate the general applicability of the technique by forming arrays of cuprous oxide (Cu(2)O), Ni, Pt, Pd, and Co exhibiting long-range order with the 18 nm hexagonal periodicity of the protein openings. This protein-based approach to electrochemical nanofabrication should permit the creation of a wide variety of two-dimensional inorganic structures. PMID- 15826096 TI - Near-field photochemical imaging of noble metal nanostructures. AB - The sub-diffraction imaging of the optical near-field in nanostructures, based on a photochemical technique, is reported. A photosensitive azobenzene-dye polymer is spin coated onto lithographic structures and is subsequently irradiated with laser light. Photoinduced mass transport creates topographic modifications at the polymer film surface that are then measured with atomic force microscopy (AFM). The AFM images correlate with rigorous theoretical calculations of the near-field intensities for a range of different nanostructures and illumination polarizations. This approach is a first step toward additional methods for resolving confined optical near fields, which can augment scanning probe methodologies for high spatial resolution of optical near fields. PMID- 15826097 TI - Inelastic effects on the transport properties of alkanethiols. AB - Using first-principles approaches, we investigate local heating and the inelastic contribution to the current for various alkanethiols sandwiched between metal electrodes. In the absence of good heat dissipation into the bulk electrodes, we find that the local temperature of the alkanethiols is relatively insensitive to their length. This is due to the rates of heating and cooling processes scaling similarly with length. On the other hand, when considering heat dissipation into the bulk electrodes, the local temperature of alkanethiols decreases as their length increases. We also find that the inelastic scattering profile displays an odd-even effect with length which compares well with experimental results. This effect is due to the alternating direction of the CH(3) group motion with respect to current flow with increasing C atoms in the chain and is very sensitive to the structure of the carbon-sulfur-gold bond. Inelastic scattering profiles can therefore help illuminate the bonding configuration of molecules to metallic surfaces. PMID- 15826098 TI - Photopatterning of actin filament structures. AB - We report a general strategy for spatiotemporal control of actin polymerization in vitro using photoactivatable actin. Caged actin was synthesized by chemically modified lysine residues on monomeric actin and released with focused ultraviolet (UV) illumination. Epifluorescence microscopy revealed nucleation and elongation of individual actin filaments (8 nm in diameter) after localized release of caged actin. We also used this strategy to generate branched filament structures by releasing caged actin in the presence of actin binding proteins. Controlled self assembly of actin filaments represents a versatile "bottom-up" technique for constructing structural building blocks and functional templates for nanoscience applications. PMID- 15826099 TI - Molecular self-assembly of "nanowires"and "nanospools" using active transport. AB - Mastering supramolecular self-assembly to a similar degree as nature has achieved on a subcellular scale is critical for the efficient fabrication of complex nanoscopic and mesoscopic structures. We demonstrate that active, molecular-scale transport powered by biomolecular motors can be utilized to drive the self assembly of mesoscopic structures that would not form in the absence of active transport. In the presented example, functionalized microtubules transported by surface-immobilized kinesin motors cross-link via biotin/streptavidin bonds and form extended linear and circular mesoscopic structures, which move in the presence of ATP. The self-assembled structures are oriented, exhibit large internal strains, and are metastable while the biomolecular motors are active. The integration of molecular motors into the self-assembly process overcomes the trade-off between stability and complexity in thermally activated molecular self assembly. PMID- 15826100 TI - Nanowire single-electron memory. AB - We demonstrate storage of electrons in semiconductor nanowires epitaxially grown from Au nanoparticles. The nanowires contain multiple tunnel junctions (MTJs) of InP barriers and InAs quantum dots designed such that the metal seed particles act as storage nodes. By positioning a second nanowire close to the seed particle it is possible to detect tunneling of individual electrons through the MTJ at 4.2 K. A strong memory effect is observed in the detector current when sweeping the writing voltage. PMID- 15826101 TI - Nanowire probes for high resolution combined scanning electrochemical microscopy atomic force microscopy. AB - We describe a method for the production of nanoelectrodes at the apex of atomic force microscopy (AFM) probes. The nanoelectrodes are formed from single-walled carbon nanotube AFM tips which act as the template for the formation of nanowire tips through sputter coating with metal. Subsequent deposition of a conformal insulating coating, and cutting of the probe end, yields a disk-shaped nanoelectrode at the AFM tip apex whose diameter is defined by the amount of metal deposited. We demonstrate that these probes are capable of high-resolution combined electrochemical and topographical imaging. The flexibility of this approach will allow the fabrication of nanoelectrodes of controllable size and composition, enabling the study of electrochemical activity at the nanoscale. PMID- 15826102 TI - Surfactant-assisted synthesis of water-soluble and biocompatible semiconductor quantum dot micelles. AB - We report a simple, rapid approach to synthesize water-soluble and biocompatible fluorescent quantum dot (QD) micelles by encapsulation of monodisperse, hydrophobic QDs within surfactant/lipid micelles. Analyses of UV-vis and photo luminescence spectra, along with transmission electron microscopy, indicate that the water-soluble semiconductor QD micelles are monodisperse and retain the optical properties of the original hydrophobic QDs. The QD micelles were shown to be biocompatible and exhibited little or no aggregation when taken up by cultured rat hippocampal neurons. PMID- 15826103 TI - Inhibition of the acetycholine esterase-stimulated growth of Au nanoparticles: nanotechnology-based sensing of nerve gases. AB - The acetylcholine esterase, AChE, mediated hydrolysis of acetylthiocholine (1) yields a reducing agent thiocholine (2) that stimulates the catalytic enlargement of Au NP seeds in the presence of AuCl(4)(-). The reductive enlargement of the Au NPs is controlled by the concentration of the substrate (1) and by the activity of the enzyme. The catalytic growth of the Au NPs is inhibited by 1,5-bis(4 allyldimethylammoniumphenyl)pentane-3-one dibromide (3) or by diethyl p nitrophenyl phosphate (paraoxon; 4), thus enabling a colorimetric test for AChE inhibitors. The colorimetric assay was also developed on glass supports. PMID- 15826104 TI - High-yield plasma synthesis of luminescent silicon nanocrystals. AB - Light emission from silicon based on quantum confinement in nanoscale structures has sparked intense research into this field ever since its discovery about 15 years ago. A barrier to the widespread utilization of luminescent silicon nanocrystals in such diverse application areas as optoelectronics, solid-state lighting for general illumination, or fluorescent agents for biological applications has been the lack of a simple, high-yield synthesis approach. Here we report a scaleable single-step synthesis process for luminescent silicon nanocrystals based on a low-pressure nonthermal plasma. PMID- 15826106 TI - Enhanced gas sensing by individual SnO2 nanowires and nanobelts functionalized with Pd catalyst particles. AB - The sensing ability of individual SnO(2) nanowires and nanobelts configured as gas sensors was measured before and after functionalization with Pd catalyst particles. In situ deposition of Pd in the same reaction chamber in which the sensing measurements were carried out ensured that the observed modification in behavior was due to the Pd functionalization rather than the variation in properties from one nanowire to another. Changes in the conductance in the early stages of metal deposition (i.e., before metal percolation) indicated that the Pd nanoparticles on the nanowire surface created Schottky barrier-type junctions resulting in the formation of electron depletion regions within the nanowire, constricting the effective conduction channel and reducing the conductance. Pd functionalized nanostructures exhibited a dramatic improvement in sensitivity toward oxygen and hydrogen due to the enhanced catalytic dissociation of the molecular adsorbate on the Pd nanoparticle surfaces and the subsequent diffusion of the resultant atomic species to the oxide surface. PMID- 15826105 TI - Six-helix bundles designed from DNA. AB - We present a designed cyclic DNA motif that consists of six DNA double helices that are connected to each other at two crossover sites. DNA double helices with 10.5 nucleotide pairs per turn facilitate the programming of DNA double crossover molecules to form hexagonally symmetric arrangements when the crossover points are separated by seven or fourteen nucleotide pairs. We demonstrate by atomic force microscopy well-formed arrays of hexagonal six-helix bundle motifs both in 1D and in 2D. PMID- 15826107 TI - Conformal anodic oxidation of aluminum thin films. AB - Membrane-based synthesis, also called template synthesis, is a very general approach used to prepare arrays of nanomaterials with monodispersed geometrical features. The most commonly used porous templates are track-etched polycarbonate and porous anodic alumina membranes. Common to all these templates is the fact that the pores are perpendicular to the surface of the membrane. Here, a novel approach is presented, where the pores are synthesized parallel to the surface of the membrane. For the first time, the anodic oxidation of an aluminum thin film is performed laterally, i.e., parallel to the surface of the substrate, instead of perpendicular as usually done. For low anodic oxidation voltages (between 3 and 5 V) we obtain highly regular and ordered pore arrays, at least over a few hundred nanometers length, with a minimum pore size of approximately 3 to 4 nm. With such porous alumina structures, the controlled in-plane organization of arrays of template-grown nanowires and carbon nanotubes for reproducible device fabrication should be much easier. PMID- 15826108 TI - Catalytic solid-phase seeding of silicon nanowires by nickel nanocrystals in organic solvents. AB - Colloidal nickel (Ni) nanocrystals were used to direct the synthesis of crystalline silicon (Si) nanowires in an organic solvent. The reaction temperatures ranged from 400 degrees C to 520 degrees C with pressures from 14.3 to 23.4 MPa, conditions that are well above the critical point of the solvent. The Ni nanocrystals play two roles in the synthesis: (1) Ni catalyzes the decomposition of the silicon precursors, i.e., arylsilanes, alkylsilanes, and trisilane, to silicon; (2) Ni nanocrystals induce silicon crystallization through the solid-phase alloying of Si in the Ni seeds. We call this nanowire growth mechanism supercritical fluid-solid-solid (SFSS) synthesis. PMID- 15826109 TI - Continuous synthesis of gold nanoparticles in a microreactor. AB - A continuous flow microreactor was used for the synthesis of gold nanoparticles (5 to 50 nm) directly from a gold salt (HAuCl(4)) and a reducing agent (ascorbic acid). Experimental parameters were optimized to obtain narrow size distributions, which were at average two times narrower than those obtained in a conventional synthesis. Additionally, two approaches, i.e., elevation of pH during reaction and hydrophobization of internal reactor surfaces were tested to suppress reactor fouling. PMID- 15826110 TI - Three-helix bundle DNA tiles self-assemble into 2D lattice or 1D templates for silver nanowires. AB - We present a DNA nanostructure, the three-helix bundle (3HB), which consists of three double helical DNA domains connected by six immobile crossover junctions such that the helix axes are not coplanar. The 3HB motif presents a triangular cross-section with one helix lying in the groove formed by the other two. By differential programming of sticky-ends, 3HB tiles can be arrayed in two distinct lattice conformations: one-dimensional filaments and two-dimensional lattices. Filaments and lattices have been visualized by high-resolution, tapping mode atomic force microscopy (AFM) under buffer. Their dimensions are shown to be in excellent agreement with designed structures. We also demonstrate an electroless chemical deposition for fabricating metallic nanowires templated on self assembled filaments. The metallized nanowires have diameters down to 20 nm and display Ohmic current-voltage characteristic. PMID- 15826111 TI - Penetrating the oxide barrier in situ and separating freestanding porous anodic alumina films in one step. AB - A simple method for penetrating the barrier layer of an anodic aluminum oxide (AAO) film and for detaching the AAO film from residual Al foil was developed by reversing the bias voltage in situ after the anodization process is completed. With this technique, we have been able to obtain large pieces of free-standing AAO membranes with regular pore sizes of sub-10 nm. By combining Ar ion milling and wetting enhancement processes, Au nanowires were grown in the sub-10 nm pores of the AAO films. Further scaling down of the pore size and extension to the deposition of nanowires and nanotubes of materials other than Au should be possible by further optimizing this procedure. PMID- 15826112 TI - Electric-field assisted growth and self-assembly of intrinsic silicon nanowires. AB - Electric-field assisted growth and self-assembly of intrinsic silicon nanowires, in-situ, is demonstrated. The nanowires are seen to respond to the presence of a localized DC electric field set up between adjacent MEMS structures. The response is expressed in the form of improved nanowire order, alignment, and organization while transcending a gap. This process provides a simple yet reliable method for enhanced control over intrinsic one-dimensional nanostructure placement and handling. PMID- 15826113 TI - Immunotargeted nanoshells for integrated cancer imaging and therapy. AB - Nanoshells are a novel class of optically tunable nanoparticles that consist of a dielectric core surrounded by a thin gold shell. Based on the relative dimensions of the shell thickness and core radius, nanoshells may be designed to scatter and/or absorb light over a broad spectral range including the near-infrared (NIR), a wavelength region that provides maximal penetration of light through tissue. The ability to control both wavelength-dependent scattering and absorption of nanoshells offers the opportunity to design nanoshells which provide, in a single nanoparticle, both diagnostic and therapeutic capabilities. Here, we demonstrate a novel nanoshell-based all-optical platform technology for integrating cancer imaging and therapy applications. Immunotargeted nanoshells are engineered to both scatter light in the NIR enabling optical molecular cancer imaging and to absorb light, allowing selective destruction of targeted carcinoma cells through photothermal therapy. In a proof of principle experiment, dual imaging/therapy immunotargeted nanoshells are used to detect and destroy breast carcinoma cells that overexpress HER2, a clinically relevant cancer biomarker. PMID- 15826114 TI - Enrichment of single-walled carbon nanotubes by diameter in density gradients. AB - The bulk enrichment and separation of single-walled carbon nanotubes (SWNTs) by diameter has been achieved through ultracentrifugation of DNA-wrapped SWNTs in aqueous density gradients. The separation is identified by the visual formation of colored bands of SWNTs in the density range of 1.11-1.17 g cm(-3). The optical absorbance spectra of the separated SWNTs indicate that SWNTs of decreasing diameter are increasingly more buoyant. This nondestructive and scalable separation strategy is expected to impact the fields of molecular electronics, optoelectronics, and sensing where SWNTs of a monodisperse band gap are essential. PMID- 15826115 TI - Periodic DNA nanotemplates synthesized by rolling circle amplification. AB - Rolling circle amplification (RCA) is an elegant biochemical method by which long single-stranded DNA molecules with a repeating sequence motif can be readily synthesized. In RCA, small circular single-stranded oligonucleotides serve as templates for the polymerization of the complementary strand. A DNA polymerase with an efficient strand displacement activity can copy the circular template without stopping. This results in a long DNA strand with periodic sequence. We here demonstrate that this method, using DNA recognition and biotin-streptavidin binding, provides a simple procedure for DNA-directed nanoscale organization of matter. As an example, a 74 nucleotide (nt) long circular DNA molecule is amplified into a sequence-periodic single strand with a length up to several micrometers. Hybridization of this long periodic DNA template to the biotinylated complement of the sequence motif results in a long DNA duplex with a periodic arrangement of biotin binding sites. On this duplex, streptavidin-coated particles can be organized into one-dimensional arrays. The resulting DNA constructs are characterized by gel electrophoresis and atomic force microscopy. PMID- 15826116 TI - Nanowire array gratings with ZnO combs. AB - Diffraction gratings are mainly manufactured by mechanical ruling, interference lithography, or resin replication, which generally require expensive equipment, complicated procedures, and a stable environment. We describe the controlled growth of self-organized microscale ZnO comb gratings by a simple one-step thermal evaporation and condensation method. The ZnO combs consist of an array of very uniform, perfectly aligned, evenly spaced and long single-crystalline ZnO nanowires or nanobelts with periods in the range of 0.2 to 2 microm. Diffraction experiments show that the ZnO combs can function as a tiny three-beam divider that may find applications in miniaturized integrated optics such as three-beam optical pickup systems. PMID- 15826117 TI - Programmable DNA self-assemblies for nanoscale organization of ligands and proteins. AB - We demonstrate the precise control of periodic spacing between individual protein molecules by programming the self-assembly of DNA tile templates. In particular, we report the application of two self-assembled periodic DNA structures, two dimensional nanogrids, and one-dimensional nanotrack, as template for programmable self-assembly of streptavidin protein arrays with controlled density. PMID- 15826118 TI - Complexes of DNA bases and gold clusters Au3 and Au4 involving nonconventional N H...Au hydrogen bonding. AB - DNA base-gold interactions are studied theoretically at the DFT level using Au(3) and Au(4) clusters as simple catalytic models for Au particles. The bonding between DNA bases and gold clusters occurs via the anchoring of a Au atom to the N or O atoms of the bases. In the most stable planar base-Au(3) complexes, the Au N or Au-O anchor bonds are reinforced by N-H...Au bonds. The mechanism of formation of these nonconventional H-bonds is discussed. PMID- 15826119 TI - Magnetomechanical detection of the specific activities of endonucleases by cantilevers. AB - Thiolated nucleic acids 1 or 2 are immobilized on Au-coated cantilevers and hybridized with the complementary nucleic acids 1a or 2a associated with magnetic particles. The duplexes 1/1a or 2/2a include specific sequences for the scission by Apa I or Mse I, respectively. The cantilevers positioned in a flow cell are subjected to an external magnetic field, leading to the deflection of the cantilevers. Upon the specific scission of the DNA duplexes by Apa I or Mse I, the magnetic particles are disconnected from the cantilevers leading to their retraction to the rest position. The deflection/retraction of the cantilevers are followed by a conventional atomic force microscope optical detection system. PMID- 15826120 TI - Fabrication of large number density platinum nanowire arrays by size reduction lithography and nanoimprint lithography. AB - Large number density Pt nanowires with typical dimensions of 12 microm x 20 nm x 5 nm (length x width x height) are fabricated on planar oxide supports. First sub 20 nm single crystalline silicon nanowires are fabricated by size reduction lithography, and then the Si nanowire pattern is replicated to produce a large number of Pt nanowires by nanoimprint lithography. The width and height of the Pt nanowires are uniform and are controlled with nanometer precision. The nanowire number density is 4 x 10(4) cm(-1), resulting in a Pt surface area larger than 2 cm(2) on a 5 x 5 cm(2) oxide substrate. Bimodal nanowires with different width have been generated by using a Pt shadow deposition technique. Using this technique, alternating 10 and 19 nm wide nanowires are produced. PMID- 15826121 TI - Photoconductivity spectra of single-carbon nanotubes: implications on the nature of their excited States. AB - We have measured the photoconductivity excitation spectra of individual semiconducting carbon nanotubes incorporated as the channel of field-effect transistors. In addition to the pronounced resonance that correlates with the second van Hove transition (E(22)) in semiconducting carbon nanotubes, a weaker sideband at about 200 meV higher energy is observed. Electronic structure calculations that include electron-phonon coupling indicate that the spectra originate from the simultaneous excitation of an exciton (main resonance) and a C C bond stretching phonon (sideband). The spectral features are not compatible with an interband interpretation of the excitation involved. PMID- 15826122 TI - The catalytic nanodiode: gas phase catalytic reaction generated electron flow using nanoscale platinum titanium oxide Schottky diodes. AB - Forty microampere current was generated on a platinum-titanium dioxide Schottky diode during the platinum catalyzed steady-state oxidation of carbon monoxide at 80 degrees C. For reaction events that produced four CO(2) molecules, three injected electrons were collected in a diode comprising a 5 nm thick platinum and a 150 nm titanium dioxide film. The electron injection flux depends on the thickness of the platinum and the titanium dioxide diode properties as well as the conditions of the catalytic reactions. PMID- 15826123 TI - Transparent and flexible carbon nanotube transistors. AB - We report the fabrication of transparent and flexible transistors where both the bottom gate and the conducting channel are carbon nanotube networks of different densities and Parylene N is the gate insulator. Device mobilities of 1 cm(2) V( 1) s(-1) and on/off ratios of 100 are obtained, with the latter influenced by the properties of the insulating layer. Repetitive bending has minor influence on the characteristics, with full recovery after repeated bending. The operation is insensitive to visible light and the gating does not influence the transmission in the visible spectral range. PMID- 15826124 TI - Failure of the vapor-liquid-solid mechanism in Au-assisted MOVPE growth of InAs nanowires. AB - We report the temperature dependence of the Au-assisted growth of InAs nanowires in MOVPE. Extensive studies of the growth of such nanowires have attributed growth to the so-called vapor-liquid-solid (VLS) mechanism, with a liquid Au-In alloy particle. We assert here that growth is instead assisted by a solid particle and does not occur at all when the particle is a liquid. Thus the temperature range of InAs nanowire growth is limited by the melting of the Au-In alloy. Comparison with growth of InAs nanowires in the same system assisted by a layer of SiO(x) is used to support this conclusion. PMID- 15826125 TI - Regulating self-assembly of spherical oligomers. AB - In multistep reactions, stability of intermediates is critical to the rate of product formation and a significant factor in generating kinetic traps. The capsid protein of cowpea chlorotic mottle virus (CCMV) can be induced to assemble into spherical particles of 30, 60, and 90 dimers. Based on examining assembly kinetics and reaction end points, we find that formation of uniform, ordered structures is not always a result of reactions that reach equilibrium. Equilibration or, alternatively, kinetic trapping can be identified by a straightforward analysis. Altering the assembly path of "spherical" particles is a means of controlling the distribution of products, which has broad applicability to self-assembly reactions. PMID- 15826126 TI - Novel method for site-controlled surface nanodot fabrication by ion beam synthesis. AB - By using a Ga FIB system to spatially control the implantation of Ga into SiO(2) followed by vacuum annealing, we have fabricated self-assembled surface Ga nanodots with a high degree of control of nucleation location. The morphology of the Ga nanodots is closely related to Ga dose, showing a critical dose needed for nucleation that results in Ga nanodot formation just below the surface, while at higher doses Ga nanodots form on the surface as metallic Ga droplets. Possible applications include defining nucleation sites for subsequent growth, use as Ga source for GaN or GaAs quantum dots, or as catalyst for nanowire growth. PMID- 15826128 TI - Probing the effects of conjugation path on the electronic transmission through single molecules using scanning tunneling microscopy. AB - A systematic study of the relationship between the molecular structure of a series of thiol end-capped oligo-phenylenevinylenes (OPVs) and the coherent electronic transmission at the single molecule level was measured by scanning tunneling microscopy (STM). This reveals a significant change in the electronic transparency of various OPV derivatives due to the insertion of a methylene spacer group or due to nitro group substitution. Apparently, changes in the conjugation path through the central benzene ring from para to meta substitution does not have a profound effect on the electronic transparency of the molecules. PMID- 15826127 TI - Manipulation and real-time electrical detection of individual bacterial cells at electrode junctions: a model for assembly of nanoscale biosystems. AB - Biological cells are complex objects that have the potential to act as templates for the subsequent construction of nanoscale structures. We demonstrate the ability to controllably and reversibly manipulate individual, live bacterial cells across micron-sized electrical gaps, and to detect bridging directly through changes in the electrical response. Our model system, Bacillus mycoides, is a rod-shaped bacterium approximately 800 nm wide and 5 microm long, similar in size and shape to many inorganic nanowires. PMID- 15826129 TI - Nanomagnetic planar magnetic resonance microscopy "lens". AB - The achievement of three-dimensional atomic resolution magnetic resonance microscopy remains one of the main challenges in the visualization of biological molecules. The prospects for single spin microscopy have come tantalizingly close due to the recent developments in sensitive instrumentation. Despite the single spin detection capability in systems of spatially well-isolated spins, the challenge that remains is the creation of conditions in space where only a single spin is resonant and detected in the presence of other spins in its natural dense spin environment. We present a nanomagnetic planar design where a localized Angstrom-scale point in three-dimensional space is created above the nanostructure with a nonzero minimum of the magnetic field magnitude. The design thereby represents a magnetic resonance microscopy "lens" where potentially only a single spin located in the "focus" spot of the structure is resonant. Despite the presence of other spins in the Angstrom-scale vicinity of the resonant spin, the high gradient magnetic field of the "lens" renders those spins inactive in the detection process. PMID- 15826130 TI - Equilibrium and nonequilibrium transport of oxygen in carbon nanotubes. AB - The equilibrium and nonequilibrium transport of O(2) through open-ended, hydrogen terminated, single-walled carbon nanotubes is examined using classical molecular dynamics simulations. It is found in both cases that the O(2) forms well-defined layers around the nanotube interior and/or exterior, and that molecular transport approaches normal mode-diffusion as the nanotube diameter increases. The interactions between the O(2) and the nantubes are stronger than that among the O(2), and this difference increases as the nanotube diameter decreases. PMID- 15826131 TI - Third-harmonic generation from single gold nanoparticles. AB - We report the first observation of third-harmonic signals from individual gold colloids down to 40 nm diameter. Excited with 1-ps pulses at 1500 nm, the colloids generate 500-nm light, close to the plasmon resonance. The third harmonic intensity varies as the square of the colloid surface area. Although weak, the third-harmonic signals of gold labels as small as 15 nm in diameter are expected to be accessible with 100-fs pulses. They could be used in microscopy for single-biomolecule tracking. PMID- 15826132 TI - Detection limits for nanoscale biosensors. AB - We examine through analytical calculations and finite element simulations how the detection efficiency of disk and wire-like biosensors in unmixed fluids varies with size from the micrometer to nanometer scales. Specifically, we determine the total flux of DNA-like analyte molecules on a sensor as a function of time and flow rate for a sensor incorporated into a microfluidic system. In all cases, sensor size and shape profoundly affect the total analyte flux. The calculations reveal that reported femtomolar detection limits for biomolecular assays are very likely an analyte transport limitation, not a signal transduction limitation. We conclude that without directed transport of biomolecules, individual nanoscale sensors will be limited to picomolar-order sensitivity for practical time scales. PMID- 15826133 TI - Comment on "gold nanoshells improve single nanoparticle molecular sensors". PMID- 15826137 TI - Virus crystals as nanocomposite scaffolds. AB - The generation of long-range three-dimensional nanoscopic patterns is a major goal in materials chemistry. Here we report a strategy for creating such systems using virus crystals as scaffolds which can be infiltrated with metal specifically palladium and platinum. The inorganic component effectively packs within the porous macromolecular crystal architecture, providing a route for patterning these materials on the nanometer length scale. To verify the quality of the metal infiltration, SEM-EDX was used to determine the homogeneous distribution of metal across the crystal, and TEM was used to confirm that the metal was confined within the porous structure of the crystal. PMID- 15826138 TI - Study of quasi-monodisperse In2O3 nanocrystals: synthesis and optical determination. AB - In this communication, we report a successful synthesis of quasi-monodisperse In2O3 nanocrystals with high crystallinity in a high-temperature organic solution. The average size of nanocrystals can be tuned using a dynamic injection technique. TEM and XRD investigations indicate that each nanocrystal is a single crystal. The optical determination implies that the photoluminescence behavior of these In2O3 nanocrystals is different from that of the bulk, probably due to the combination of weak quantum-confinement-effects and the nature of high crystallinity in nanocrystals. PMID- 15826140 TI - Electrochemical behavior of the tris(pyridine)-Cu funnel complexes: an overall induced-fit process involving an entatic state through a supramolecular stress. AB - The electrochemical behavior of the tris(pyridine) calix[6]arene Cu adducts is unique as compared to that of most classical Cu complexes in a strain-free environment. The presence of MeCN buried inside the cavity is a prerequisite for a quasi-reversible behavior in a dynamic mode. The CV behavior assisted by simulation outlines that the coordination adaptability of the Cu(II)/Cu(I) redox states is completely reversed, with a Td geometry enforced at either redox states. Hence, the supramolecular control of the Cu coordination by a protein like pocket determines the dynamics of the electron transfer process, its thermodynamics, and the kinetics of the reorganizational barrier and generates a preorganized state for oxidation. This redox behavior corresponds to an overall induced-fit process generating a truly entatic highly oxidizing Cu(II) state through a protein-like strain by involvement of the secondary coordination sphere. PMID- 15826139 TI - Hangman salophens. AB - We report here a modular approach for the construction of a new class of compounds, the Hangman salophens. In the Hangman motif, an acid-base functionality "hangs" over the face of a redox cofactor. In contrast to more synthetically intractable porphyrin-based Hangman systems, Hangman salophens permit the facile control of their proton and redox properties for the study of the proton-coupled electron transfer (PCET) activation of small molecules. By investigating the catalase-like disproportionation of H2O2, we show that the presence (1) of a strong proton-donating hanging group (i.e., carboxylic acid) and (2) of electron-donating groups on the redox-active salen imparts significant catalytic activity for the O-O bond activation of small molecule substrates. The contribution of the new ligand framework to furthering our understanding of how PCET can be implemented in the design of active/selective catalysts will be discussed. PMID- 15826141 TI - Intramolecular C-H/C-D exchange in cofacially stacked polyfluorenes via electron induced bond activation. AB - EPR studies in hexamethylphosphoramide, along with DFT studies, reveal that the one-electron reduction of pi-stacked polyfluorenes containing two, three, and four fluorene units leads to the corresponding anion radicals where the odd electron is located exclusively on an outside fluorene moiety. These anion radicals disproportionate to form small concentrations of diamagnetic dianions, wherein there is a nonclassical pz overlap that crosses the C2 axis. The presence of the two extra electrons activates the C-H bonds in the ortho positions, and concomitant overlap of the p-orbitals involving adjacent fluorene moieties results in intramolecular hydrogen exchange at temperatures as low as 90 K. PMID- 15826142 TI - Selective and catalytic arylation of N-phenylpyrrolidine: sp3 C-H bond functionalization in the absence of a directing group. AB - We herein describe our studies on arylation of N-phenylpyrrolidine, which led to the development of a new transformation for the direct and selective arylation of sp3 C-H bonds in the absence of a directing group. In this method, Ru(H)2(CO)(PCy3)3 4 was used as the catalyst, and preliminary mechanistic studies suggested that Ru(Ph)(I)(CO)(PCy3)2 5 is the key intermediate of the catalytic cycle. A large kinetic isotope effect (kH/kD = 5.4) was observed, which supports the proposal that C-H bond metalation is the slow step. Preliminary examination of the substrate scope showed that in addition to N-phenylpyrrolidine, N-methyl- and N-benzylpyrrolidine, as well as N-benzoylpyrrolidine, were arylated under the reaction conditions. PMID- 15826143 TI - Na5(In4S)(InS4)3.6H2O, a zeolite-like structure with unusual SIn4 tetrahedra. AB - A hydrated sodium indium sulfide, Na5(In4S)(InS4)3.6H2O, has been prepared by hydrothermal synthesis. This material contains a tetrahedral sulfur site coordinated to four trivalent indium ions, an unusual bonding pattern not previously observed in open framework chalcogenides. The structure is related to the perovskite (CaTiO3) type with simultaneous substitutions of Ti by SIn4, O by InS4, and Ca2+ by [Na5(H2O)6]5+. It is a wide-gap semiconductor and shows photocatalytic activity under UV light for hydrogen generation from aqueous solution without use of any cocatalyst. PMID- 15826145 TI - Unusually short RNA sequences: design of a 13-mer RNA that selectively binds and recognizes theophylline. AB - RNA plays critical roles in numerous biological processes and constitutes valuable therapeutic targets. RNA is significant not only for its roles in transmitting the genetic code but also for its enzymatic functions in ribozymes and in peptide bond formation in ribosomes. Recent studies have shown that RNAs containing as few as 22 nucleotides can be key elements in cellular functions. This suggests the possibility of using short RNAs as regulatory elements. Here, we show that ligand recognition and selectivity by RNA molecules can occur with only the presence of a binding pocket and as few as six additional scaffolding nucleotides holding the binding pocket in place. A 13-mer RNA truncation of a 33 mer aptamer for theophylline preserves the ability to bind to theophylline and to discriminate against the structurally similar compound caffeine. The truncated aptamer retains nearly all of the same structural elements in its binding site as those present in the original aptamer. This is the first demonstration of selective ligand binding by a 13-mer RNA. PMID- 15826146 TI - Activation of high-T(C) ferromagnetism in Mn2+-doped ZnO using amines. AB - We report the discovery that high-TC ferromagnetism in manganese-doped ZnO (Mn2+:ZnO) can be activated by amine binding and calcination. The activation of ferromagnetism is attributed to the incorporation of uncompensated p-type dopants into the ZnO lattice upon amine calcination, a process that has substantial precedence in the literature surrounding p-type ZnO. The experimental observations are consistent with a microscopic mechanism involving formation of bound magnetic polarons upon introduction of p-type dopants into Mn2+:ZnO. These results clearly demonstrate that Mn2+:ZnO ferromagnetism is critically sensitive to defects other than the magnetic dopants themselves, offering some insight into the diversity of experimental observations reported previously for this material. PMID- 15826144 TI - Bio-orthogonal affinity purification of direct kinase substrates. AB - Protein phosphorylation is a major mechanism of post-translational protein modification used to control cellular signaling. A challenge in phosphoproteomics is to identify the direct substrates of each protein kinase. Herein, we describe a chemical strategy for delivery of a bio-orthogonal affinity tag to the substrates of an individual protein kinase. The kinase of interest is engineered to transfer a phosphorothioate moiety to phosphoacceptor hydroxyl groups on direct substrates. In a second nonenzymatic step, the introduced phosphorothioate is alkylated with p-nitrobenzylmesylate (PNBM). Antibodies directed against the alkylated phosphorothioate epitope recognize these labeled substrates, but not alkylation products of other cellular nucleophiles. This strategy is demonstrated with Cdk1/cyclinB substrates using ELISA, western blotting, and immunoprecipitation in the context of whole cell lysates. PMID- 15826147 TI - Template-free directional growth of single-walled carbon nanotubes on a- and r plane sapphire. AB - We report high-throughput growth of highly aligned single-walled carbon nanotube arrays on a-plane and r-plane sapphire substrates. This is achieved using chemical vapor deposition with ferritin as the catalyst. The nanotubes are aligned normal to the [0001] direction for growth on the a-plane sapphire. They are typically tens of micrometers long, with a narrow diameter distribution of 1.34 +/- 0.30 nm. In contrast, no orientation was achieved for growth on the c plane and m-plane sapphire, or when Fe films, instead of ferritin, were used as the catalyst. Such orientation control is likely related to the interaction between carbon nanotubes and the sapphire substrate, which is supported by the observation that when a second layer of nanotubes was grown, they followed the gas flow direction. These aligned nanotube arrays may enable the construction of integrable and scalable nanotube devices and systems. PMID- 15826148 TI - Photoswitchable orientational patterns of confined domains in monolayers. AB - We report here a photoswitch process that involves collective molecular reorientation in a monolayer of an azobenzene derivative. Using polarized light we force the transition between two clearly distinguishable orientational mesoscopic configurations that can be monitored by reflection optical microscopy. A model that combines thermodynamic and kinetic arguments is proposed, and it is able to reproduce both the two states and the mechanism involved in the transition. We conclude that the phenomenon reported here is essentially different from the usual electric-field-induced molecular alignment often found in liquid crystalline materials and devices. Instead, it involves a photoexcitation concomitant with an H-aggregation process. PMID- 15826149 TI - Chemical confirmation of a pentavalent phosphorane in complex with beta phosphoglucomutase. AB - This communication reports the X-ray crystal structure of the alpha-d-galactose-1 phosphate complex with that of Lactococcus lactis beta-phosphoglucomutase (beta PGM) crystallized in the presence of Mg2+ cofactor and the enzyme-to-phosphorus ratio determined by protein and phosphate analyses of the crystalline complex. The 1:1 ratio determined for this complex was compared to the 1:2 ratio determined for the crystals of beta-PGM grown in the presence of substrate and Mg2+ cofactor. This result verifies the published structure assignment of this latter complex as the phosphorane adduct formed by covalent bonding between the active site Asp8 carboxylate to the C(1)phosphorus of the beta-glucose 1,6 bisphosphate ligand and rules out the proposal of a beta-PGM-glucose-6-phosphate 1-MgF3- complex. PMID- 15826150 TI - Doubly catalytic sensing of HIV-1-related CCR5 sequence in prokaryotic cell-free translation system using riboregulator-controlled luciferase activity. AB - A molecular-beacon-type riboregulator (mRNA) was applied to multiply catalytic gene sensing. It consists of a reporter gene for firefly protein luciferase and, upstream thereof, a regulator hairpin domain composed of an RBS/anti-RBS stem (RBS = ribosome binding site) and a loop which is complementary to the target. The hairpin and, hence, the RBS are rendered open upon binding of a target oligonucleotide of the human CC chemokine receptor 5 sequence in a prokaryotic cell-free translation system (10 muL) to ignite ribosomal catalytic translation, or transcription/translation when using a DNA form of the probe, to produce luciferase, which is assayed by a catalytic chemiluminescence reaction. The sensing, using an unmodified RNA or even dsDNA as a probe with a chemiluminescence output, is thus doubly catalytic or amplifiable with a sensitivity at 95%). We also obtained evidence that 2 can form 1 under mild conditions, and that 2 reacts with nucleophiles, such as methanol, by conjugate addition. PMID- 15826172 TI - Prosthetic heme modification during halide ion oxidation. Demonstration of chloride oxidation by horseradish peroxidase. AB - Myeloperoxidase (MPO), eosinophil peroxidase (EPO), and chloroperoxidase can oxidize iodide, bromide, and chloride, but most peroxidases, including the prototypical horseradish peroxidase (HRP), reportedly only oxidize iodide and, in some cases, bromide. We report here that incubation of HRP with Br(-) and H(2)O(2) at acidic pH results in both bromination of monochlorodimedone and modification of the heme group. Mass spectrometry indicates that the heme 2- and 4-vinyl groups are modified by either replacement of a vinyl hydrogen by a bromide or addition of HOBr to give a bromohydrin. These reactions do not occur if protein-free heme and Br(-) are co-incubated with H(2)O(2) or if the HRP reaction is carried out at pH 7. Surprisingly, similar prosthetic heme modifications occur in incubations of HRP with H(2)O(2) and Cl(-). A mechanism is proposed involving oxidation of Br(-) or Cl(-) to give HOBr or HOCl, respectively, followed by addition to a vinyl group. In the reaction with Cl(-), a meso-chloro heme adduct is also formed. This first demonstration of Cl(-) oxidation by HRP, and the finding that prosthetic heme modification occurs when Br(-) or Cl(-) is oxidized in the absence of a cosubstrate, show that only modest tuning is required to achieve the unique chloride oxidation activity of MPO and EPO. The results raise the question of how the prosthetic hemes of MPO and EPO, whose function is to produce oxidized halide species, escape modification. PMID- 15826173 TI - Single-molecule tracing on a fluidic microchip for quantitative detection of low abundance nucleic acids. AB - Here, we report a method capable of quantitative detection of low-abundance DNA/RNA molecules by incorporating confocal fluorescence spectroscopy, molecular beacons, and a molecular-confinement microfluidic reactor. By using a combination of ac and dc fields via a trio of 3-D electrodes in the microreactor, we are able to precisely direct the transport of individual molecules to a minuscule laser focused detection volume ( approximately 1 fL). A burst of fluorescence photons is detected whenever a molecular beacon-target hybrid flows through the detection region, and the amount of targets can be directly quantified according to the number of recorded single-molecule flow-through events. This assay consumes only attomoles of molecular probes and is able to quantitatively detect subpicomolar DNA targets. A measurement time of less than 2 min is sufficient to complete the detection. PMID- 15826174 TI - Mechanistic studies on the binding of nitric oxide to a synthetic heme-thiolate complex relevant to cytochrome p450. AB - The synthetic heme-thiolate complex (SR) in methanol binds nitric oxide (k(on) = (2.7 +/- 0.2) x10(6) M(-)(1) s(-)(1) at 25 degrees C) to form SR(NO). The binding of NO to the SR complex in a noncoordinating solvent, such as toluene, was found to be almost 3 orders of magnitude faster than that in methanol. The activation parameters DeltaH(), DeltaS(), and DeltaV() for the formation of SR(NO) in methanol are consistent with the operation of a limiting dissociative mechanism, dominated by dissociation of methanol in SR(MeOH). In the presence of an excess of NO, the formation of SR(NO) is followed by subsequent slower reactions. The substantially negative activation entropy and activation volume values found for the second observed reaction step support an associative mechanism which involves attack of a second NO molecule on the thiolate ligand in the initially formed SR(NO) complex. The following slower reactions are strongly accelerated by a large excess of NO or by the presence of NO(2)(-) in the SR/NO reaction mixture. They can be accounted for in terms of dynamic equilibria between higher nitrogen oxides (NO(x)()) and reactive SR species, which lead to the formation of a nitrosyl-nitrite complex of SR(Fe(II)) as the final product. This finding is clearly supported by laser flash photolysis studies on the SR/NO reaction mixture, which do not reveal simple NO photolabilization from SR(Fe(III))(NO), but rather involve the generation of at least three photoinduced intermediates decaying with different rate constants to the starting material. The species formed along the proposed reaction pathways were characterized by FTIR and EPR spectroscopy. The results are discussed in terms of their relevance for the biological function of cytochrome P450 enzymes and in context of results for the reaction of NO with imidazole- and thiolate-ligated iron(III) hemoproteins. PMID- 15826175 TI - Succinct synthesis of beta-amino acids via chiral isoxazolines. AB - beta-Amino acids are important synthetic targets due to their presence in a wide variety of natural products, pharmaceutical agents, and mimics of protein structural motifs. While beta-amino acids containing geminal substitution patterns have enormous potential for application in these contexts, synthetic challenges to the stereoselective preparation of this class of compound have thus far limited more complete studies. We present here a straightforward method employing chiral isoxazolines as key intermediates to access five different beta amino acid structural types with excellent selectivity. Of particular note is the use of this approach to prepare highly substituted cis-beta-proline analogues. The ready access to these diversely substituted compounds is expected to facilitate future studies of the structure and function of this important class of molecules. PMID- 15826176 TI - Enantioselective cyanosilylation of ketones catalyzed by a chiral oxazaborolidinium ion. AB - The chiral oxazaborolidinium salt 1 (X = TfO) is an excellent catalyst for the cyanosilylation of methyl ketones promoted by trimethylsilyl cyanide and diphenylmethyl phosphine oxide as co-reactants (to generate Ph(2)MePOTMS(N=C:) as a reactive intermediate). The face selectivity of this reaction parallels that previously observed for the corresponding reaction of aldehydes. A unifying and rational mechanistic explanation is provided for these enantioselective reactions. Evidence is presented to support the importance of alpha-C-H...O hydrogen bonding, pi,pi-interaction of the complexed ketonic carbonyl with the mexyl group of 1, and an early transition state for high enantioselectivity. The cyanosilylation reaction described herein provides access to many useful chiral compounds. PMID- 15826177 TI - Chemistry of the diazeniumdiolates: Z right harpoon over left harpoon E isomerism. AB - Here, we explore the chemistry of the previously undocumented E form of diazeniumdiolates having the structure R(1)R(2)NN(O)=NOR(3). Reported crystallographic studies have uniformly revealed the Z configuration, and our attempts to observe a Z --> E conversion through thermal equilibration or photochemical means have, until now, consistently failed to reveal a significant amount of a second conformer. As a typical example, the NMR spectrum of trimethyl derivative Me(2)NN(O)=NOMe revealed no evidence for a second configuration. Electronic structure calculations attribute this finding to a prohibitively high interconversion barrier of approximately 40 kcal/mol. A similar result was obtained when we considered the case of R(1) = Me = R(3) and R(2) = H at the same levels of theory. However, when MeHNN(O)=NOMe was ionized by dissociating the N-H bond, the barrier was calculated to be lower by approximately 20 kcal/mol, with the E form of the anion being favored over Z. This circumstance suggested that an E isomer might be isolable if a Z anion were formed and given sufficient time to assume the E configuration, then quenched by reaction with an electrophile to trap and neutralize the E form and restore the putatively high interconversion barrier. Consistent with this prediction, basifying iPrHNN(O)=NOCH(2)CH(2)Br rapidly led to a six-membered heterocycle that was crystallographically characterized as containing the -N(O)=NO- functional group in the E configuration. The results suggest an approach for generating pairs of Z and E diazeniumdiolates for systematic comparison of the rates at which the individual isomers release bioactive NO and of other physicochemical determinants of their biomedical utility. PMID- 15826178 TI - Ruthenium(salen)-catalyzed aerobic oxidative desymmetrization of meso-diols and its kinetics. AB - Chiral (nitrosyl)ruthenium(salen) complexes were found to be efficient catalysts for aerobic oxidative desymmetrization of meso-diols under photoirradiation to give optically active lactols. The scope of the applicability of this reaction ranges widely from acyclic diols to mono-cyclic diols, although fine ligand tuning of the ruthenium(salen) complexes was required to attain high enantioselectivity (up to 93% ee). In particular, the nature of the apical ligand was found to affect not only enantioselectivity but also kinetics of the desymmetrization reaction. Spectroscopic analysis of the oxidation disclosed that irradiation of visible light is indispensable not only for dissociation of the nitrosyl ligand but also for single electron transfer from the alcohol-bound ruthenium ion to dioxygen. PMID- 15826179 TI - Dependence of transition state structure on substrate: the intrinsic C-13 kinetic isotope effect is different for physiological and slow substrates of the ornithine decarboxylase reaction because of different hydrogen bonding structures. AB - Ornithine decarboxylase is the first and the rate-controlling enzyme in polyamine biosynthesis; it decarboxylates l-ornithine to form the diamine putrescine. We present calculations performed using a combined quantum mechanical and molecular mechanical (QM/MM) method with the AM1 semiempirical Hamiltonian for the wild type ornithine decarboxylase reaction with ornithine (the physiological substrate) and lysine (a "slow" substrate) and for mutant E274A with ornithine substrate. The dynamical method is variational transition state theory with quantized vibrations. We employ a single reaction coordinate equal to the carbon carbon distance of the dissociating bond, and we find a large difference between the intrinsic kinetic isotope effect for the physiological substrate, which equals 1.04, and that for the slow substrate, which equals 1.06. This shows that, contrary to a commonly accepted assumption, kinetic isotope effects on slow substrates are not always good models of intrinsic kinetic isotope effects on physiological substrates. Furthermore, analysis of free-energy-based samples of transition state structures shows that the differences in kinetic isotope effects may be traced to different numbers of hydrogen bonds at the different transition states of the different reactions. PMID- 15826180 TI - Structure, function, and inhibition along the reaction coordinate of CTX-M beta lactamases. AB - CTX-M enzymes are an emerging group of extended spectrum beta-lactamases (ESBLs) that hydrolyze not only the penicillins but also the first-, second-, and third generation cephalosporins. Although they have become the most frequently observed ESBLs in certain areas, there are few effective inhibitors and relatively little is known about their detailed mechanism. Here we describe the X-ray crystal structures of CTX-M enzymes in complex with different transition-state analogues and beta-lactam inhibitors, representing the enzyme as it progresses from its acylation transition state to its acyl enzyme complex to the deacylation transition state. As the enzyme moves along this reaction coordinate, two key catalytic residues, Lys73 and Glu166, change conformations, tracking the state of the reaction. Unexpectedly, the acyl enzyme complex with the beta-lactam inhibitor cefoxitin still has the catalytic water bound; this water had been predicted to be displaced by the unusual 7alpha-methoxy of the inhibitor. Instead, the 7alpha-group appears to inhibit by preventing the formation of the deacylation transition state through steric hindrance. From an inhibitor design standpoint, we note that the best of the reversible inhibitors, a ceftazidime like boronic acid compound, binds to CTX-M-16 with a K(i) value of 4 nM. When used together in cell culture, this inhibitor reversed cefotaxime resistance in CTX-M-producing bacteria. The structure of its complex with CTX-M enzyme and the structural view of the reaction coordinate described here provide templates for inhibitor design and intervention to combat this family of antibiotic resistance enzymes. PMID- 15826181 TI - Glycopeptides related to beta-endorphin adopt helical amphipathic conformations in the presence of lipid bilayers. AB - A series of glycosylated endorphin analogues designed to penetrate the blood brain barrier (BBB) have been studied by circular dichroism and by 2D-NMR in the presence of water; TFE/water; SDS micelles; and in the presence of both neutral and anionic bicelles. In water, the glycopeptides showed only nascent helix behavior and random coil conformations. Chemical shift indices and nuclear Overhauser effects (NOE) confirmed helices in the presence of membrane mimics. NOE volumes provided distance constraints for molecular dynamics calculations used to provide detailed backbone conformations. In all cases, the glycopeptides were largely helical in the presence of membrane bilayer models (micelles or bicelles). Plasmon waveguide resonance (PWR) studies showed hen egg phosphatidyl choline (PC) bilayers produce amphipathic helices laying parallel to the membrane surface, with dissociation constants (K(D)) in the low nanomolar to micromolar concentration range. Two low-energy states are suggested for the glycosylated endorphin analogues, a flexible aqueous state and a restricted membrane bound state. Strong interactions between the glycopeptide amphipaths and membranes are crucial for penetration of the BBB via an endocytotic mechanism (transcytosis). PMID- 15826182 TI - Spectroscopic and computational studies of Ni superoxide dismutase: electronic structure contributions to enzymatic function. AB - Ni-containing superoxide dismutase (NiSOD) is the most recently discovered member of the class of metalloenzymes that detoxify the superoxide radical in aerobic organisms. In this study, we have employed a variety of spectroscopic and computational methods to probe the electronic structure of the NiSOD active site in both its oxidized (NiSOD(ox), possessing a low-spin (S = (1)/(2)) Ni(3+) center) and reduced (NiSOD(red), containing a diamagnetic Ni(2+) center) states. Our experimentally validated computed electronic-structure description for NiSOD(ox) reveals strong sigma-bonding interactions between Ni and the equatorial S/N ligands, which give rise to intense charge-transfer transitions in the near UV region of the absorption spectrum. Resonance Raman (rR) spectra obtained with laser excitation in this region exhibit two features at 349 and 365 cm(-)(1) that are assigned to Ni-S(Cys) stretching modes. The NiSOD(red) active site also exhibits a high degree of metal-ligand bond covalency as well as filled/filled pi interactions between Ni and S/N orbitals, which serve to adjust the redox potential of the Ni(2+) center. Comparison of our computational results for NiSOD(red) with those obtained in parallel studies of synthetic [NiS(2)N(2)] complexes reveals that the presence of an anionic N-donor ligand is crucial for promoting metal-based (versus S-based) oxidation of the active site. The implications of our electronic-structure descriptions with respect to the function of NiSOD are discussed, and a comparison of M-S(Cys) bonding in NiSOD and other metalloenzymes with sulfur ligation is provided. PMID- 15826183 TI - Terephthalamide derivatives as mimetics of helical peptides: disruption of the Bcl-x(L)/Bak interaction. AB - A series of Bcl-x(L)/Bak antagonists, based on a terephthalamide scaffold, was designed to mimic the alpha-helical region of the Bak peptide. These molecules showed favorable in vitro activities in disrupting the Bcl-x(L)/Bak BH3 domain complex (terephthalamides 9 and 26, K(i) = 0.78 +/- 0.07 and 1.85 +/- 0.32 microM, respectively). Extensive structure-affinity studies demonstrated a correlation between the ability of terephthalamide derivatives to disrupt Bcl x(L)/Bak complex formation and the size of variable side chains on these molecules. Treatment of human HEK293 cells with the terephthalamide derivative 26 resulted in disruption of the Bcl-x(L)/Bax interaction in whole cells with an IC(50) of 35.0 microM. Computational docking simulations and NMR experiments suggested that the binding cleft for the BH3 domain of the Bak peptide on the surface of Bcl-x(L) is the target area for these synthetic inhibitors. PMID- 15826185 TI - Spin-crossover dendrimers: generation number-dependent cooperativity for thermal spin transition. AB - Poly(benzyl ether) dendrons having a focal triazole unit (Gntrz: trz = triazole; n = generation number = 0-2) were found to react with (MeSO(3))(2)Fe to form dendritic coordination polymers ([Fe(Gntrz)(3)](MeSO(3))(2).2H(2)O) that undergo the thermal spin transition. When the generation number of the dendritic unit was larger (n = 0 --> 1 --> 2), the average degree of polymerization (D(p) = 20 --> 10 --> 3) and spin-crossover temperature (T(c) = 335 --> 315 --> 300 K) of the resulting polymer were lower. However, the abruptness of the spin transition was not monotonically dependent on the generation number; (G1trz)Fe exhibited an abrupt spin transition with a temperature width of only 10 K, while the smallest and largest members of the (Gntrz)Fe family both displayed a rather broad spin transition temperature width (30 (n = 0) and 50 K (n = 2)). X-ray diffraction and calorimetric analyses indicated the presence of a discotic columnar core-shell assembly with a crystal lattice best occupied by a C(3)(v)() symmetric array of medium-sized (G1trz)Fe. PMID- 15826184 TI - Substrate oxidation by copper-dioxygen adducts: mechanistic considerations. AB - A series of copper-dioxygen adducts [{Cu(II)(MePY2)(R)}(2)(O(2))](B(C(6)F(5))(4))(2) (1(R)()), systematically varying in their electronic properties via ligand pyridyl donor substituents (R = H, MeO, and Me(2)N), oxidize a variety of substrates with varying C-H or O-H bond dissociation enthalpies. Detailed mechanistic studies have been carried out, including investigation of 1(R)() thermodynamic redox properties, 1(R)() tetrahydrofuran (THF) and N,N'-dimethylaniline (DMA) oxidation kinetics (including analyses of substrate dicopper binding equilibria), and application of mechanistic probes (N-cyclopropyl-N-methylaniline (CMA) and (p-methoxyphenyl)-2,2 dimethylpropanol (MDP)), which can distinguish if proton-coupled electron transfer (PCET) processes proceed through concerted electron-transfer proton transfer (ETPT) or consecutive electron-transfer proton-transfer (ET/PT) pathways. The results are consistent with those of previous complementary studies; at low thermodynamic driving force for substrate oxidation, an ET/PT is operable, but once ET (i.e., substrate one-electron oxidation) becomes prohibitively uphill, the ETPT pathway occurs. Possible differences in coordination structures about 1(Me)()()2(N)()/1(MeO)() compared to those of 1(H)() are also used to rationalize some of the observations. PMID- 15826186 TI - "Majority-rules" operative in chiral columnar stacks of C3-symmetrical molecules. AB - C(3)-symmetrical disks 1, preorganized by acylated 2,2'-bipyridine-3,3'-diamine moieties and decorated with nine identical chiral, lipophilic tails, aggregate into a dynamic helix in apolar solvents. The aggregates, previously shown to be governed by the "sergeants-and-soldiers" principle when mixed with achiral analogues, are now also revealed to obey the "majority-rules" effect, a phenomenon not earlier observed in nonpolymers. Our experimental circular dichroism data can be accurately described with a recently developed theory. A fit of the theory to the experimental results shows that the mismatch penalty, i.e., the free energy of a monomer present in a helix of its nonpreferred screw sense (0.94 kJ/mol), is about 8 times lower than the penalty for a helix reversal (7.8 kJ/mol). This corresponds well to our vision of the supramolecular architecture of the disks. PMID- 15826187 TI - Copper(I) 1,2,4-triazolates and related complexes: studies of the solvothermal ligand reactions, network topologies, and photoluminescence properties. AB - One-pot solvothermal treatments of organonitriles, ammonia, and Cu(II) salts yielded Cu(I) and 3,5-disubstituted 1,2,4-triazolates. The organic triazolate components were derived from copper-mediated oxidative cycloaddition of nitriles and ammonia, in which a key intermediate 1,3,5-triazapentadienate was isolated as [Cu(II)(4-pytap)(2)] (4-Hpytap = 2,4-di(4-pyridyl)-1,3,5-triazapentadiene) via controlled solvothermal conditions. This intermediate could also be synthesized by Ni(II)-mediated reactions; however, the final triazoles were obtained only when Cu(II) was employed. Therefore, the reaction mechanism of these reactions was elucidated as follows: nitrile was first attacked by ammonia to form the amidine, which further reacted with another nitrile or self-condensed to yield 1,3,5-triazapentadiene, which was coordinated to two Cu(II) ions in its deprotonated form. A two-electron oxidation of the 1,3,5-triazapentadienate mediated by two Cu(II) ions gave one triazolate and Cu(I) cations. Other in situ ligand reactions, such as C-C bond cleavage and hydrolysis, were also found for the nitriles under these solvothermal conditions. Another remarkable feature of these crystalline Cu(I) triazolates is their simple, typical 3- or 4-connected network topologies. The self-assembly of these nets is presumably controlled by steric hindrance, which is subsequently applied to the rational design of the close-packed 2D networks [Cu(I)(tz)](infinity) and [Ag(I)(tz)](infinity) (Htz = 1,2,4-triazole), as well as the porous 3D network [Cu(I)(etz)](infinity) (Hetz = 3,5-diethyl-1,2,4-triazole). The interesting photoluminescence properties of these coinage d(10) metal complexes were also investigated. PMID- 15826188 TI - Stepwise and dramatic enhancement of anion recognition with a triple-site receptor based on the calix[4]arene framework using two different cationic effectors. AB - Synthesis and binding behavior of a novel multi-responsive host 1, in which two esters, two polyether moieties, two urea sites, and two bipyridine units as ion binding sites are arranged on the calix[4]arene skeleton, is reported. 1 recognizes Na(+) and Ag(+) simultaneously and quantitatively and captures an anionic guest. The ability of 1 to recognize anions, including CF(3)SO(3)(-) and BF(4)(-), remarkably increases in a stepwise manner using Na(+) and Ag(+) as effectors. The enhancement of the K(a) eventually reaches factors of 1500 and 2000 for NO(3)(-) and CF(3)SO(3)(-), respectively, in the presence of both Na(+) and Ag(+), compared to the free 1. The regulation of binding of multiple ligands may be applicable to multistep cascade systems for the amplification of molecular events, and further studies in this field could provide insight applicable to more advanced molecular devices. PMID- 15826189 TI - Crystalline polymorph selection and discovery with polymer heteronuclei. AB - The discovery and selective production of crystalline polymorphs, an outstanding problem in solid-state chemistry, is of great importance industrially in, for example, the manufacture of pharmaceuticals and pigments. Despite considerable efforts, no reliable method exists to produce all of the stable polymorphs of a given compound. Herein, we report methodology to control the phenomenon of crystal polymorphism through the use of diverse libraries of polymer heteronuclei including both commercially available polymers and combinatorially synthesized cross-linked polymers. This new approach for exploring polymorph space offers the advantage of high throughput crystallization to discover multiple polymorphs combined with the ability to selectively produce a given form from a single solvent and temperature condition by simply varying the nature of the polymer substrate. This technique is successfully demonstrated on the pharmaceuticals acetaminophen, sulfamethoxazole, and carbamazepine and on the pharmaceutical intermediate 5-methyl-2-[(2-nitrophenyl)amino]-3-thiophenecarbonitrile (ROY). High throughput screening, accomplished by optical microscopy and Raman spectroscopy, identified the selective production of the two stable polymorphs of acetaminophen and all six stable forms of ROY. Furthermore, one new form of carbamazepine and two new forms of sulfamethoxazole were discovered; in these cases, single crystals were obtained enabling the structural characterization of two new tetramorphic systems. PMID- 15826190 TI - Stereochemistry of hexenyl radical cyclizations with tert-butyl and related large groups: substituent and temperature effects. AB - The long held notion that hexenyl radicals bearing large substituents on the radical carbon cyclize to give 1,2-trans-substituted cyclopentanes is experimentally disproved by study of the radical cyclization of an assortment of simple and complex substrates coupled with careful product analysis and rigorous assignment of configurations. X-ray studies and syntheses of authentic samples establish that the published assignments for cis- and trans-1-tert-butyl-2 methylcyclopentane must be reversed. The original assignment based on catalytic hydrogenation of 1-tert-butyl-2-methylenecyclopentane was compromised by migration of the double bond prior to hydrogenation. The cyclization of 1-tert butylhexenyl radical is moderately cis selective, and the selectivity is increased by geminal substitution on carbon 3. This selectivity trend is general and extends to relatively complex substrates. It has allowed Ihara to reduce the complexity of an important class of round trip radical cyclizations to make linear triquinanes to the point where two tricyclic products-cis-syn-cis and cis anti-cis-account for about 80% of the products. However, the further increase in selectivity that was proposed by lowering the temperature is shown to be an artifact of the analysis methods and is not correct. This work solidifies "1,2 cis selectivity" in cyclizations of 1-subsituted hexenyl radicals as one of the most general stereochemical trends in radical cyclizations. PMID- 15826191 TI - Scandium(III) catalysis of transimination reactions. Independent and constitutionally coupled reversible processes. AB - Sc(OTf)(3) efficiently catalyzes the self-sufficient transimination reaction between various types of C=N bonds in organic solvents, with turnover frequencies up to 3600 h(-)(1) and rate accelerations up to 6 x 10(5). The mechanism of the crossover reaction in mixtures of amines and imines is studied, comparing parallel individual reactions with coupled equilibria. The intrinsic kinetic parameters for isolated reactions cannot simply be added up when several components are mixed, and the behavior of the system agrees with the presence of a unique mediator that constitutes the core of a network of competing reactions. In mixed systems, every single amine or imine competes for the same central hub, in accordance with their binding affinity for the catalyst metal ion center. More generally, the study extends the basic principles of constitutional dynamic chemistry to interconnected chemical transformations and provides a step toward dynamic systems of increasing complexity. PMID- 15826192 TI - Kinetic stabilization of an oligomeric protein by a single ligand binding event. AB - Protein native state stabilization imposed by small molecule binding is an attractive strategy to prevent the misfolding and misassembly processes associated with amyloid diseases. Transthyretin (TTR) amyloidogenesis requires rate-limiting tetramer dissociation before misassembly of a partially denatured monomer ensues. Selective stabilization of the native TTR tetramer over the dissociative transition state by small molecule binding to both thyroxine binding sites raises the kinetic barrier of tetramer dissociation, preventing amyloidogenesis. Assessing the amyloidogenicity of a TTR tetramer having only one amyloidogenesis inhibitor (I) bound is challenging because the two small molecule binding constants are generally not distinct enough to allow for the exclusive formation of TTR.I in solution to the exclusion of TTR.I(2) and unliganded TTR. Herein, we report a method to tether one fibril formation inhibitor to TTR by disulfide bond formation. Occupancy of only one of the two thyroxine binding sites is sufficient to inhibit tetramer dissociation in 6.0 M urea and amyloidogenesis under acidic conditions by imposing kinetic stabilization on the entire tetramer. The sufficiency of single occupancy for stabilizing the native state of TTR provides the incentive to search for compounds displaying striking negative binding cooperativity (e.g., K(d1) in nanomolar range and K(d2) in the micromolar to millimolar range), enabling lower doses of inhibitor to be employed in the clinic, mitigating potential side effects. PMID- 15826193 TI - Role of conformation and electronic structure in the chemistry of ground and excited state o-pyrazolylphenylnitrenes. AB - The chemistry of 2-(1-pyrazolyl)- (2a) and 2-[1-(3,5 dimethylpyrazolyl]phenylnitrene (2b) has been studied in EtOH solution at room temperature, in EtOH glasses at 90 K, and in Ar matrices at 12 K. These nitrenes were chosen as suitable models for clarifying the mechanism of intramolecular reactions because attack at the pyrazole ring can occur according to different modes and the asymmetry of the substituent gives rise to different conformations. Detailed DFT and CASSCF/CASPT2 studies on the conformation and decay paths of both spin states of the nitrenes have been carried out. Ring expansion to dehydroazepines (via benzoazirines) is calculated to be competitive in both nitrenes, but in the dimethyl derivative, 2b, attack onto the N lone pair (which is made more nucleophilic by the methyl groups) is favored. The higher barriers (by 4-8 kcal/mol) in singlet 2a cause 60-70% of this nitrene to decay by intersystem crossing to the triplet. Thus, the seemingly straightforward formation of benzo-fused heterocycles through intramolecular attack of the pyrazoline N lone pair by the singlet phenylnitrene can only overcome ring expansion and intermolecular reactions under favorable circumstances. The comparatively persistent triplet nitrenes are characterized in matrices, and the yields of photocyclization products (mainly pyrazolo[1,5-a]benzimidazole (7) from 2a and 5,6-dihydropyrazolo[1,5-a]quinoxaline (8) from 2b) are shown to depend on the preferred conformation of the starting azide and nitrene. PMID- 15826194 TI - Superacidifiers: assessing the activation and the mode of charge transmission of the extraordinary electron-withdrawing SO2CF3 and S(O)(=NSO2CF3)CF3 substituents in carbanion stabilization. AB - We report on a structural (multinuclear NMR), thermodynamic (pK(a)), and kinetic (Marcus intrinsic reactivity) study of the ionization of benzylic carbon acids activated by an exocyclic (alpha) SO(2)CF(3) group and SO(2)CF(3) or S(O)(=NSO(2)CF(3))CF(3) in the para position of the phenyl ring. The latter exerts an enormous acidifying effect of ca. 8 pK units as compared with 4-H benzyltriflone in Me(2)SO solution, (corresponding to remarkably high Hammett sigma values sigma(p) approximately 1.35, sigma(p)(-) approximately 2.30). In considering the origin of this effect, important information was derived in comparing medium effects on pK(a)'s for NO(2), SO(2)CF(3), and S(O)(=NSO(2)CF(3))CF(3) activated carbon acids. Highly contrasting behavior was thus induced by H(2)O --> Me(2)SO transfer, with a large decrease in acidity of alpha-nitro activated carbon acids but a large increase in acidity of alpha SO(2)CF(3) analogues, leading to remarkable inversions in C-H acidity. These results support the view that in the case of the triflones the carbanion negative charge resides for the most part at the exocyclic Calpha carbon, implying a major role of a polarizability effect. (1)H, (13)C, and (19)F NMR data fully support this proposal. Most importantly, the intrinsic reactivity (log k(0)) positioning 9 and 10 on the Marcus scale for carbon acids could be kinetically measured in 50%H(2)O-50%Me(2)SO; for 9, log k(0) = 3.80 and for 10, log k(0) = 4.20. Such high log k(0) values correspond to low intrinsic barriers which can only be reconciled on the basis of minimum electronic and structural reorganization in formation of the conjugate carbanions. This further emphasizes polarization as the predominant mechanistic mode of charge stabilization in these species. PMID- 15826195 TI - Studies of an enneanuclear manganese single-molecule magnet. AB - The reaction of [Mn(3)O(O(2)CMe)(6)(py)(3)] with the tripodal ligand H(3)thme (1,1,1-tris(hydroxymethyl)ethane) affords the enneanuclear complex [Mn(9)O(7)(O(2)CCH(3))(11)(thme)(py)(3)(H(2)O)(2)] 1.1MeCN.1Et(2)O. The metallic skeleton of complex 1 comprises a series of 10 edge-sharing triangles that describes part of an idealized icosahedron. Variable temperature direct current (dc) magnetic susceptibility data collected in the 1.8-300 K temperature range and in fields up to 5.5 T were fitted to give a spin ground state of S = (17)/(2) with an axial zero-field splitting parameter D = -0.29 cm(-)(1). Ac susceptibility studies indicate frequency-dependent out-of-phase signals below 4 K and an effective barrier for the relaxation of the magnetization of U(eff) = 27 K. Magnetic measurements of single crystals of 1 at low temperature show time- and temperature-dependent hysteresis loops which contain steps at regular intervals of field. Inelastic neutron scattering (INS) studies on complex 1 confirm the S = (17)/(2) ground state and analysis of the INS transitions within the zero-field split ground state leads to determination of the axial anisotropy, D = -0.249 cm(-)(1), and the crystal field parameter, B(4)(0) = 7(4) x 10(-)(6) cm(-)(1). Frequency domain magnetic resonance spectroscopy (FDMRS) determined the same parameters as D = -0.247 cm(-)(1) and B(4)(0) = 4.6 x 10(-)(6) cm(-)(1). DFT calculations are fully consistent with the experimental findings of two Mn(II) and four Mn(III) ions "spin up" and three Mn(IV) ions "spin down" resulting in the S = (17)/(2) spin ground state of the molecule, with D = -0.23 cm(-)(1) and U = 26.2 K. PMID- 15826196 TI - The influence of sequence context and length on the kinetics of DNA duplex formation from complementary hairpins possessing (CNG) repeats. AB - The formation of unusual structures during DNA replication has been invoked for gene expansion in genomes possessing triplet repeat sequences, CNG, where N = A, C, G, or T. In particular, it has been suggested that the daughter strand of the leading strand partially dissociates from the parent strand and forms a hairpin. The equilibrium between the fully duplexed parent:daugter species and the parent:hairpin species is dependent upon their relative stabilities and the rates of reannealing of the daughter strand back to the parent. These stabilities and rates are ultimately influenced by the sequence context of the DNA and its length. Previous work has demonstrated that longer strands are more stable than shorter strands and that the identity of N also influences the thermal stability [Paiva, A. M.; Sheardy, R. D. Biochemistry 2004, 43, 14218-14227]. Here, we show that the rate of duplex formation from complementary hairpins is also sequence context and length dependent. In particular, longer duplexes have higher activation energies than shorter duplexes of the same sequence context. Further, [(CCG):(GGC)] duplexes have lower activation energies than corresponding [(CAG):(GTC)] duplexes of the same length. Hence, hairpins formed from long CNG sequences are more thermodynamically stable and have slower kinetics for reannealing to their complement than shorter analogues. Gene expansion can now be explained in terms of thermodynamics and kinetics. PMID- 15826197 TI - Poly-p-phenylene phosphine/polyaniline alternating copolymers: electronic delocalization through phosphorus. AB - Phosphorus-containing poly(N-arylaniline)s and related polymer model compounds have been prepared. The spectroscopic and electronic properties of the materials were investigated via UV-vis-NIR spectroscopy and cyclic voltammetry. PPPP-PANI copolymers containing p-phenylene diamine units in the polymer backbone have electronic and spectroscopic properties characteristic of aromatic substituted p phenylene diamines. Copolymers containing -(-C(6)H(4)-P-C(6)H(4)-P-C(6)H(4)-)- linkages between nitrogen centers show evidence for weak electronic delocalization along the polymer chain. The electrochemical and spectroscopic properties support strong electronic delocalization in copolymers containing -(-P C(6)H(4)-N-C(6)H(4)-)- repeat units. The presence of a single diphenylphosphine bridge between nitrogen centers provides an efficient mode of electronic delocalization between nitrogen centers. PPPP oxide-PANI copolymers and related polymer model compounds were also prepared and investigated. The resemblance of PPPP oxide-PANI copolymers to isolated p-phenylene diamines or triarylamines suggests electronic isolation of the amine fragments in the polymer. The conversion of phosphorus(III) phosphines to phosphorus(V) phosphine oxides inhibits electronic delocalization through phosphorus, further supporting delocalization of the lone pair of electrons on phosphorus in PPPP-PANI copolymers. PPPP-PANI copolymers are a new type of pi-conjugated polymer with low oxidation potentials and electronic delocalization through phosphorus along the polymer chain. PMID- 15826198 TI - Crotylsilane reagents in the synthesis of complex polyketide natural products: total synthesis of (+)-discodermolide. AB - An efficient, highly convergent stereocontrolled synthesis of (+)-discodermolide has been achieved with 2.1% overall yield (27 steps longest linear sequence). The absolute stereochemistry of the C1-C6 (12), C7-C14 (13), and C15-C24 (11) subunits was introduced using asymmetric crotylation methodology. Key elements of the synthesis include the use of hydrozirconation-cross-coupling methodology for the construction of C13-C14 (Z)-olefin, acetate aldol reaction to construct the C6-C7 bond and install the C7 stereocenter with high levels of 1,5-anti stereoinduction, and the use of palladium-mediated sp(2)-sp(3) cross-coupling reaction to join the advanced fragments, which assembled the carbon framework of discodermolide. PMID- 15826199 TI - Catalytic enantioselective construction of all-carbon quaternary stereocenters: synthetic and mechanistic studies of the C-acylation of silyl ketene acetals. AB - With the aid of an appropriate chiral catalyst, acyclic silyl ketene acetals react with anhydrides to furnish 1,3-dicarbonyl compounds that bear all-carbon quaternary stereocenters in good ee and yield. Mechanistic studies provide strong support for a catalytic cycle that involves activation of both the electrophile (anhydride --> acylpyridinium) and the nucleophile (silyl ketene acetal --> enolate). PMID- 15826200 TI - A general nonaqueous route to binary metal oxide nanocrystals involving a C-C bond cleavage. AB - A widely applicable solvothermal route to nanocrystalline iron, indium, gallium, and zinc oxide based on the reaction between the corresponding metal acetylacetonate as metal oxide precursor and benzylamine as solvent and reactant is presented. Detailed XRD, TEM, and Raman studies prove that, with the exception of the iron oxide system, where a mixture of the two phases magnetite and maghemite is formed, only phase pure materials are obtained, gamma-Ga(2)O(3), zincite ZnO, and cubic In(2)O(3). The particle sizes lie in the range of 15-20 nm for the iron, 10-15 nm for the indium, 2.5-3.5 nm for gallium, and around 20 nm for zinc oxide. GC-MS analysis of the final reaction solution after removal of the nanoparticles showed that the composition is rather complex consisting of more than eight different organic compounds. Based on the fact that N isopropylidenebenzylamine, 4-benzylamino-3-penten-2-one, and N-benzylacetamide were the main species found, we propose a detailed formation mechanism encompassing solvolysis of the acetylacetonate ligand, involving C-C bond cleavage, as well as ketimine and aldol-like condensation steps. PMID- 15826201 TI - MOCVD-derived highly transparent, conductive zinc- and tin-doped indium oxide thin films: precursor synthesis, metastable phase film growth and characterization, and application as anodes in polymer light-emitting diodes. AB - Four diamine adducts of bis(hexafluoroacetylacetonato)zinc [Zn(hfa)(2).(diamine)] can be synthesized in a single-step reaction. Single crystal X-ray diffraction studies reveal monomeric, six-coordinate structures. The thermal stabilities and vapor phase transport properties of these new complexes are considerably greater than those of conventional solid zinc metal-organic chemical vapor deposition (MOCVD) precursors. One of the complexes in the series, bis(1,1,1,5,5,5 hexafluoro-2,4-pentadionato)(N,N'-diethylethylenediamine)zinc, is particularly effective in the growth of thin films of the transparent conducting oxide Zn-In Sn-O (ZITO) because of its superior volatility and low melting point of 64 degrees C. ZITO thin films with In contents ranging from 40 to 70 cation % (a metastable phase) were grown by low-pressure MOCVD. These films exhibit conductivity as high as 2900 S/cm and optical transparency comparable to or greater than that of commercial Sn-doped indium oxide (ITO) films. ZITO films with the nominal composition of ZnIn(2.0)Sn(1.5)O(z)() were used in fabrication of polymer light-emitting diodes. These devices exhibit light outputs and current efficiencies almost 70% greater than those of ITO-based control devices. PMID- 15826202 TI - Cobalt(III) corroles as electrocatalysts for the reduction of dioxygen: reactivity of a monocorrole, biscorroles, and porphyrin-corrole dyads. AB - Three series of cobalt(III) corroles were tested as catalysts for the electroreduction of dioxygen to water. One was a simple monocorrole represented as (Me(4)Ph(5)Cor)Co, one a face-to-face biscorrole linked by an anthracene (A), biphenylene (B), 9,9-dimethylxanthene (X), dibenzofuran (O) or dibenzothiophene (S) bridge, (BCY)Co(2) (with Y = A, B, X, O or S), and one a face-to-face bismacrocyclic complex, (PCY)Co(2), containing a Co(II) porphyrin and a Co(III) corrole also linked by one of the above rigid spacers (Y = A, B, X, or O). Cyclic voltammetry and rotating ring-disk electrode voltammetry were both used to examine the catalytic activity of the cobalt complexes in acid media. The mixed valent Co(II)/Co(III) complexes, (PCY)Co(2), and the biscorrole complexes, (BCY)Co(2), which contain two Co(III) ions in their air-stable forms, all provide a direct four-electron pathway for the reduction of O(2) to H(2)O in aqueous acidic electrolyte when adsorbed on a graphite electrode, with the most efficient process being observed in the case of the complexes having an anthracene spacer. A relatively small amount of hydrogen peroxide was detected at the ring electrode in the vicinity of E(1/2) which was located at 0.47 V vs SCE for (PCA)Co(2) and 0.39 V vs SCE for (BCA)Co(2). The cobalt(III) monocorrole (Me(4)Ph(5)Cor)Co also catalyzes the electroreduction of dioxygen at E(1/2) = 0.38 V with the final products being an approximate 50% mixture of H(2)O(2) and H(2)O. PMID- 15826203 TI - Determination of the temperature dependence of the H-D spin-spin coupling constant and the isotope effect on the proton chemical shift for the compressed dihydride complex [CpIr(P-P)H2]2+. AB - Complex [CpIr(dmpm)H(2)](2+) (dmpm = bis(dimethylphosphino)methane) has been reported to display temperature-dependent spin-spin coupling constant ((1)J(HD)) and isotope effect on the (1)H NMR chemical shift (Deltadelta). A combined electronic structure density functional theory + quantum nuclear dynamics study is used to determine from first-principles the unusual temperature dependence of the spin-spin coupling constant. It is found that the potential energy surface describing the motion of the Ir-H(2) unit has a deeper minimum in the dihydride region and is characterized by important anharmonicities. These anomalies affect the nature of the vibrational states of the unit and are the main reason for the unusual temperature dependence of (1)J(HD) and Deltadelta. These results suggest experimental tests to identify compressed dihydride transition metal complexes. PMID- 15826204 TI - Redox-noninnocence of the S,S'-coordinated ligands in bis(benzene-1,2 dithiolato)iron complexes. AB - The electronic structures of complexes of iron containing two S,S'-coordinated benzene-1,2-dithiolate, (L)(2)(-), or 3,5-di-tert-butyl-1,2-benzenedithiolate, (L(Bu))(2)(-), ligands have been elucidated in depth by electronic absorption, infrared, X-band EPR, and Mossbauer spectroscopies. It is conclusively shown that, in contrast to earlier reports, high-valent iron(IV) (d(4), S = 1) is not accessible in this chemistry. Instead, the S,S'-coordinated radical monoanions (L(*))(1)(-) and/or (L(Bu)(*))(1)(-) prevail. Thus, five-coordinate [Fe(L)(2)(PMe(3))] has an electronic structure which is best described as [Fe(III)(L)(L(*))(PMe(3))] where the observed triplet ground state of the molecule is attained via intramolecular, strong antiferromagnetic spin coupling between an intermediate spin ferric ion (S(Fe) = (3)/(2)) and a ligand radical (L(*))(1)(-) (S(rad) = (1)/(2)). The following complexes containing only benzene 1,2-dithiolate(2-) ligands have been synthesized, and their electronic structures have been studied in detail: [NH(C(2)H(5))(3)](2)[Fe(II)(L)(2)] (1), [N(n Bu)(4)](2)[Fe(III)(2)(L)(4)] (2), [N(n-Bu)(4)](2)[Fe(III)(2)(L(Bu))(4)] (3); [P(CH(3))Ph(3)][Fe(III)(L)(2)(t-Bu-py)] (4) where t-Bu-py is 4-tert butylpyridine. Complexes containing an Fe(III)(L(*))(L)- or Fe(III)(L(Bu))(L(Bu)(*))- moiety are [N(n-Bu)(4)][Fe(III)(2)(L(Bu))(3)(L(Bu)(*))] (3(ox)()), [Fe(III)(L)(L(*))(t-Bu-py)] (4(ox)()), [Fe(III)(L(Bu))(L(Bu)(*))(PMe(3))] (7), [Fe(III)(L(Bu))(L(Bu)(*))(PMe(3))(2)] (8), and [Fe(III)(L(Bu))(L(Bu)(*))(PPr(3))] (9), where Pr represents the n-propyl substituent. Complexes 2, 3(ox)(), 4, [Fe(III)(L)(L(*))(PMe(3))(2)] (6), and 9 have been structurally characterized by X-ray crystallography. PMID- 15826205 TI - Reversible enolization of beta-amino carboxamides by lithium hexamethyldisilazide. AB - The enolization of beta-amino carboxamides by lithium hexamethyldisilazide (LiHMDS) in THF/toluene and subsequent diastereoselective alkylation with CH(3)I are reported. In situ IR spectroscopic studies reveal that beta-amino carboxamides coordinate to LiHMDS at -78 degrees C before enolization. Comparison with structurally similar carboxamides suggests that the beta-amino group promotes the enolization. IR spectroscopic studies also show that the enolization is reversible. Efficient trapping of the enolate by CH(3)I affords full conversion to products. (6)Li and (15)N NMR spectroscopic studies reveal that lithium enolate-LiHMDS mixed dimers and trimers as well as a homoaggregated enolate are formed during the reaction. At ambient temperature, racemization of the beta-position through a putative reversible Michael addition was observed. PMID- 15826206 TI - Synthesis of quantum-sized cubic ZnS nanorods by the oriented attachment mechanism. AB - Quantum-sized ZnS nanocrystals with quasi-spherical and rod shapes were synthesized by the aging reaction mixtures containing diethylzinc, sulfur, and amine. Uniform-sized ZnS nanorods with the average dimension of 5 nm x 21 nm, along with a small fraction of 5 nm-sized quasi-spherical nanocrystals, were synthesized by adding diethylzinc to a solution containing sulfur and hexadecylamine at 125 degrees C, followed by aging at 300 degrees C. Subsequent secondary aging of the nanocrystals in oleylamine at 60 degrees C for 24 h produced nearly pure nanorods. Structural characterizations showed that these nanorods had a cubic zinc blende structure, whereas the fabrication of nanorods with this structure has been known to be difficult to achieve via colloidal chemical synthetic routes. High-resolution TEM images and reaction studies demonstrated that these nanorods are formed from the oriented attachment of quasi spherical nanocrystals. Monodisperse 5 nm-sized quasi-spherical ZnS nanocrystals were separately synthesized by adding diethylzinc to sulfur dissolved in a mixture of hexadecylamine and 1-octadecene at 45 degrees C, followed by aging at 300 degrees C. When oleic acid was substituted for hexadecylamine and all other procedures were unchanged, we obtained 10 nm-sized quasi-spherical ZnS nanocrystals, but with broad particle size distribution. These two different sized quasi-spherical ZnS nanocrystals showed different proportions of zinc blende and wurtzite crystal structures. The UV absorption spectra and photoluminescence excitation spectra of the 5 nm ZnS quasi-spherical nanocrystals and of the nanorods showed a blue-shift from the bulk band-gap, thus showing a quantum confinement effect. The photoluminescence spectra of the ZnS nanorods and quasi-spherical nanocrystals showed a well-defined excitonic emission feature and size- and shape-dependent quantum confinement effects. PMID- 15826207 TI - Unexpected behavior of the surface composition of PtRh alloys during chemical reaction. AB - The changes in the surface composition of a Pt(50)Rh(50)(100) alloy due to an ongoing 2H(2) + O(2)-->2H(2)O chemical reaction have been studied in situ. Exploiting the high-energy resolution and surface sensitivity of synchrotron radiation core-level spectroscopy it was possible to monitor the population of the two transitions metals atoms at the gas-surface interface. Sequences of fast high-resolution core-level spectra of the Rh3d(5/2), Pt4f(7/2), and O1s core levels showed a continuous exchange of atoms between the first and subsurface layers. An unexpected Pt surface enrichement was found under slightly oxidizing conditions, opposite to what found in a highly oxidizing atmosphere. PMID- 15826208 TI - Electronic configuration assignment and the importance of low-lying excited states in high-spin imidazole-ligated iron(II) porphyrinates. AB - The synthesis and characterization of six new high-spin deoxymyoglobin models (imidazole(tetraarylporphyrinato)iron(II)) are described. These have been intensively studied by temperature-dependent Mossbauer spectroscopy from 295 to 4.2 K. All complexes show a strong temperature dependence for the quadrupole splitting consistent with low-lying excited states of the same or lower multiplicity. An analysis of the data obtained in applied magnetic fields leads to the assignment of the sign of the quadrupole splitting. All model compounds as well as those of deoxymyoglobin and deoxyhemoglobin, previously studied, have a negative sign for the quadrupole splitting. Although not previously predicted, this experimental observation leads to the assignment of the ground-state electronic configuration for all high-spin imidazole-ligated iron(II) porphyrinates as (d(xz)())(2)(d(yz)())(1)(d(xy)())(1)(d(z)()()2)(1)(d(x)()()2( )(y)()()2)(1). This is a distinctly different ground-state electronic configuration from other high-spin iron(II) porphyrinates; differences in structural details for the two classes of high-spin complexes are also discussed. The apparent anomaly of differing signs for the zero-field splitting constant between previously studied model complexes and the heme proteins is addressed; the difference appears to result from the fact that the assumptions used in the spin Hamiltonian approach that has been applied to these complexes are not adequately satisfied. Structures of four of the new five-coordinate species have been determined. Core conformations in these derivatives show variation, but these and previously studied compounds reveal a limited number of conformational patterns. The bond lengths and other geometrical parameters such as porphyrin core size and iron out-of-plane displacement support a high-spin state assignment for the iron(II). PMID- 15826209 TI - Functionalization of thioctic acid-capped gold nanoparticles for specific immobilization of histidine-tagged proteins. AB - This paper presents an efficient strategy for the specific immobilization of fully functional proteins onto the surface of nanoparticles. Thioctic acid derivatized gold clusters are used as a scaffold for further stepwise modification, leading to a cobalt(II)-terminated ligand shell. A histidine tag introduced by genetic engineering into a protein is coordinated to this transition metal ion. The specific immobilization has been demonstrated for the cases of a genetically engineered horseradish peroxidase and ferredoxin-NADP(+) reductase, confirming the attachment of the fully functional proteins to the Co(II)-terminated nanointerface. The absence of nonspecific protein adsorption and the specificity of the binding site have been verified using several analogues of the enzymes without the histidine tag. PMID- 15826210 TI - Colorimetric molecularly imprinted polymer sensor array using dye displacement. AB - A colorimetric sensor array composed of seven molecularly imprinted polymers was shown to accurately identify seven different aromatic amines. The response patterns were systematically classified using linear discriminant analysis with 94% classification accuracy. Analyses of the response patterns of the analytes to the imprinted polymer array suggest that the different selectivity patterns, although subtle, appear to arise from the imprinting process. The molecular imprinting process enabled the rapid preparation of the polymers in the array from ethylene glycol dimethacrylate and methacrylic acid (80:20) in the presence of six different template molecules plus a blank nonimprinted polymer. The response of the imprinted polymer array was coupled to a colorimetric response, using a dye displacement strategy. A benzofurazan dye was selected and shown to give an accurate measure of the binding properties of the imprinted polymer array to all seven analytes. The colorimetric response also enabled the inclusion of analytes that are not spectroscopically active and were not among the original analytes that were used as template molecules. This broadens the potential utility of the imprinted polymer sensor array strategy to a wider range of analytes and applications. PMID- 15826211 TI - Evidence for d orbital aromaticity in square planar coinage metal clusters. AB - Quantitative evidence for the existence of aromaticity involving the d orbitals of transition metals is provided for the first time. The doubly bridged square planar (D(4)(h)()) coinage metal clusters (M(4)Li(2), M = Cu (1), Ag (2), and Au (3)) are characterized as aromatic by their substantial nucleus independent chemical shifts (NICS) values in the centers (-14.5, -14.1, and -18.6, respectively). Nevertheless, the participation of p orbitals in the bonding (and cyclic electron delocalization) of 1-3 is negligible. Instead, these clusters benefit strongly from the delocalization of d and to some extent s orbitals. The same conclusion applies to Tsipis and Tsipis' H-bridged D(4)(h)() Cu(4)H(4) ring (4). Canonical MO-NICS analysis of structures 1-3 shows the total diatropic d orbital contributions to the total NICS to be substantial, although the individual contributions of the five sets of filled d orbitals vary. The d orbital aromaticity of Cu(4)Li(2) also is indicated by its atomization energy, 243.2 kcal/mol, which is larger than Boldyrev's doubly (sigma and pi) aromatic Al(4)Li(2) (215.9 kcal/mol). PMID- 15826212 TI - Cross surface ambipolar charge percolation in molecular triads on mesoscopic oxide films. AB - We report on cross surface ambipolar charge percolation within a monolayer of a molecular triad adsorbed on semiconducting or insulating mesoscopic metal oxide films. The triad consists of a triphenlyamine (TPA) donor and a perylenemonoimide (PMI) acceptor connected by a bithiophene (T2) bridge. The self-assembled PMI-T2 TPA monolayer exhibits p-type or n-type conduction depending on the potential that is applied to the conducting glass (FTO) electrode supporting the oxide films. Cross surface electron transfer is turned on at around -1.24 V (vs Fc+/Fc) where the PMI moiety is electroactive. The color of the film changes from red to blue during the reduction of the PMI. By contrast, lateral hole transfer is turned on at around 0.8 V (vs Fc+/Fc) where the TPA moiety becomes electroactive. The stepwise oxidation of the T2-TPA units at 0.79 and 1.28 V (vs Fc+/Fc) is associated with a color change of the film from red to black. Cyclic voltammetric as well as chronocoulometric and spectroelectrochemical measurements were applied to determine the percolation threshold for cross surface charge transfer and the diffusion coefficients for the electron and hole hopping process. The effect of oxide surface states on the lateral charge motion was also investigated. PMID- 15826213 TI - Diffusion measurements for molecular capsules: pulse sequences effect on water signal decay. AB - Diffusion NMR and, more recently, diffusion ordered spectroscopy (DOSY) are gaining popularity as efficient tools for the characterization of supramolecular systems in solution. Here, using diffusion NMR of hydrogen-bond molecular capsules, we demonstrate that the use of different diffusion sequences may have a dramatic effect on exchanging peaks. In fact, we found that the signal decay of the water peak in [(1a)(6)(H(2)O)(8)] is monoexponential in the pulsed gradient spin-echo (PGSE) and stimulated echo (PGSTE) sequences and biexponential in the longitudinal eddy current delay (LED) and the bipolar longitudinal eddy current delay (BPLED) sequences, routinely used in modern DOSY experiments. By performing these diffusion measurements on molecular capsules, in which water is not part of the molecular capsules, we demonstrate that this phenomenon is observed only for water molecules that exchange between two sites that differ considerably in their diffusion coefficients. Degeneration of the LED or the BPLED sequences into PGSTE type sequences by shortening the te period resulted in the disappearance of the extra slow diffusing component. The origin, as well as the implications of the different results obtained from conventional diffusion sequences, such as the PGSE and PGSTE as compared with the LED and BPLED sequences generally used in DOSY experiments, are briefly discussed. PMID- 15826214 TI - Systematic control of photochemistry: the dynamics of photoisomerization of a model cyanine dye. AB - On-the-fly CASSCF nonadiabatic dynamics have been used to model the trans-cis isomerization of a model cyanine dye. Our results show that the photochemical generation of the trans versus cis product is dynamically controlled by the presence of an extended cis-trans conical intersection seam that persists along all torsional angles. This in turn suggests that the photochemistry could be completely controlled by controlling the distribution of momentum components in a wave packet excited by laser photolysis in a coherent control experiment. PMID- 15826219 TI - Vocal experimentation in the juvenile songbird requires a basal ganglia circuit. AB - Songbirds learn their songs by trial-and-error experimentation, producing highly variable vocal output as juveniles. By comparing their own sounds to the song of a tutor, young songbirds gradually converge to a stable song that can be a remarkably good copy of the tutor song. Here we show that vocal variability in the learning songbird is induced by a basal-ganglia-related circuit, the output of which projects to the motor pathway via the lateral magnocellular nucleus of the nidopallium (LMAN). We found that pharmacological inactivation of LMAN dramatically reduced acoustic and sequence variability in the songs of juvenile zebra finches, doing so in a rapid and reversible manner. In addition, recordings from LMAN neurons projecting to the motor pathway revealed highly variable spiking activity across song renditions, showing that LMAN may act as a source of variability. Lastly, pharmacological blockade of synaptic inputs from LMAN to its target premotor area also reduced song variability. Our results establish that, in the juvenile songbird, the exploratory motor behavior required to learn a complex motor sequence is dependent on a dedicated neural circuit homologous to cortico-basal ganglia circuits in mammals. PMID- 15826218 TI - The neural substrates of infant sleep in rats. AB - Sleep is a poorly understood behavior that predominates during infancy but is studied almost exclusively in adults. One perceived impediment to investigations of sleep early in ontogeny is the absence of state-dependent neocortical activity. Nonetheless, in infant rats, sleep is reliably characterized by the presence of tonic (i.e., muscle atonia) and phasic (i.e., myoclonic twitching) components; the neural circuitry underlying these components, however, is unknown. Recently, we described a medullary inhibitory area (MIA) in week-old rats that is necessary but not sufficient for the normal expression of atonia. Here we report that the infant MIA receives projections from areas containing neurons that exhibit state-dependent activity. Specifically, neurons within these areas, including the subcoeruleus (SubLC), pontis oralis (PO), and dorsolateral pontine tegmentum (DLPT), exhibit discharge profiles that suggest causal roles in the modulation of muscle tone and the production of myoclonic twitches. Indeed, lesions in the SubLC and PO decreased the expression of muscle atonia without affecting twitching (resulting in "REM sleep without atonia"), whereas lesions of the DLPT increased the expression of atonia while decreasing the amount of twitching. Thus, the neural substrates of infant sleep are strikingly similar to those of adults, a surprising finding in light of theories that discount the contribution of supraspinal neural elements to sleep before the onset of state dependent neocortical activity. PMID- 15826220 TI - Work stress, substance use, and depression among young adult workers: an examination of main and moderator effect model. AB - In this cross-sectional study, main and moderated relationships between 5 job stressors and alcohol consumption, drug use, and depression were examined using data from a community sample of 583 young adults (mean age = 23.68 years). Analyses revealed a few direct associations between high job boredom, low skill variety, and low autonomy and depression measures and heavy alcohol use. There were no direct relationships between job stress and binge drinking, alcohol consumption, drug use, or heavy drug use. In a few cases, job stress-outcome relationships were moderated by intrinsic job motivation or gender. The findings supported a specificity-of-effects hypothesis and underscored the need for examining the processes linking occupational stress to substance use and depression. PMID- 15826221 TI - The impact of job characteristics on work-to-family facilitation: testing a theory and distinguishing a construct. AB - This study used objective measures of job characteristics appended to the National Survey of Midlife Development in the United States (MIDUS), self reported job characteristics, and an individual resource characteristic (orientation toward personal growth) to test a theory of work-family facilitation. Results indicated that resource-rich jobs enable work-to-family facilitation. A higher level of work-to-family facilitation was reported by individuals in jobs with more autonomy and variety and whose jobs required greater substantive complexity and social skill. There was no support for the hypotheses that these effects would be more pronounced for individuals with higher levels of personal growth. The authors found significant differences in the strength of the associations of job characteristics with work-to-family facilitation and work-tofamily conflict, suggesting they are different constructs with distinct antecedents. PMID- 15826222 TI - Are trends in work and health conditions interrelated? A study of Swedish hospital employees in the 1990s. AB - Swedish hospital personnel were followed over an 8-year period, characterized by staff redundancies and restructuring processes. Self-rated and administrative data sets from 1994 to 2001 allowed for studying long-term consequences of organizational instability for staff health and work conditions. The aim was to identify, on a work-unit level, trends in work and health conditions and their interdependence. Regression analysis showed a downward trend in mental health and an upward trend in long-term sick leave. Increasing trends of work demands were accompanied by deteriorating mental health, and decreasing time to plan work showed the strongest association with increasing long-term sick leave. Job satisfaction and support were decreasing. A stable short-term sick leave rate over years related to lack of support. PMID- 15826223 TI - The impact of deployment length and experience on the well-being of male and female soldiers. AB - This study examined the effects of stressor duration (deployment length) and stressor novelty (no prior deployment experience) on the psychological health of male and female military personnel returning from a peacekeeping deployment. The sample consisted of men (n = 2,114) and women (n = 1,225) surveyed for symptoms of depression and posttraumatic stress. The results confirmed the hypotheses. Longer deployments and 1st-time deployments were associated with an increase in distress scores. However, the relationship between deployment length and increased distress was found only for male soldiers. The findings demonstrate the importance of considering the impact of exposure to long-term occupational stressors and confirm, in part, previous research that has demonstrated a different stress response pattern for men and women. PMID- 15826224 TI - The longitudinal effects of work-family conflict and positive spillover on depressive symptoms among dual-earner couples. AB - This study assessed longitudinal and cross-sectional relationships between work family conflict, positive spillover, and depression in a national sample of 234 dual-earner couples. The authors also assessed crossover effects (i.e., the transmission of emotions, affect, or stress from 1 member of a dyad to another) of work-family conflict and positive spillover on spouses' depression. Two general findings of the study were that (a) positive spillover has a stronger impact on depression than does work-family conflict, and (b) the effects of spouses' positive spillover were more strongly related to decreased depression than were the effects of one's own positive spillover. Significant longitudinal effects were related to the crossover of positive spillover on decreased spouse depression. PMID- 15826225 TI - Sexual versus nonsexual workplace aggression and victims' overall job satisfaction: a meta-analysis. AB - A meta-analytic approach was used to examine whether sexual and nonsexual forms of nonviolent workplace aggression (both verbal and nonverbal) share equivalent or differential relationships with victims' overall job satisfaction. When the meta-analytic comparison was restricted to all-female samples to hold victims' gender constant, nonsexual aggression was found to share a significantly stronger negative relationship with victims' overall job satisfaction than was sexual aggression. In addition, nonsexual aggression was found to share a stronger negative link with women's level of job satisfaction than with men's. PMID- 15826226 TI - Job resources buffer the impact of job demands on burnout. AB - This study tested and refined the job demands-resources model, demonstrating that several job resources play a role in buffering the impact of several job demands on burnout. A total of 1,012 employees of a large institute for higher education participated in the study. Four demanding aspects of the job (e.g., work overload, emotional demands) and 4 job resources (e.g., autonomy, performance feedback) were used to test the central hypothesis that the interaction between (high) demands and (low) resources produces the highest levels of burnout (exhaustion, cynicism, reduced professional efficacy). The hypothesis was rejected for (reduced) professional efficacy but confirmed for exhaustion and cynicism regarding 18 out of 32 possible 2-way interactions (i.e., combinations of specific job demands and resources). PMID- 15826227 TI - Multiple-object tracking is based on scene, not retinal, coordinates. AB - This study tested whether multiple-object tracking-the ability to visually index objects on the basis of their spatiotemporal history-is scene based or image based. Initial experiments showed equivalent tracking accuracy for objects in 2-D and 3-D motion. Subsequent experiments manipulated the speeds of objects independent of the speed of the scene as a whole. Results showed that tracking accuracy was influenced by object speed but not by scene speed. This held true whether the scene underwent translation, zoom, rotation, or even combinations of all 3 motions. A final series of experiments interfered with observers' ability to see a coherent scene by moving objects at different speeds from one another and by distorting the perception of 3-D space. These reductions in scene coherence led to reduced tracking accuracy, confirming that tracking is accomplished using a scene-based, or allocentric, frame of reference. PMID- 15826228 TI - Early, involuntary top-down guidance of attention from working memory. AB - Four experiments explored the interrelations between working memory, attention, and eye movements. Observers had to identify a tilted line amongst vertical distractors. Each line was surrounded by a colored shape that could be precued by a matching item held in memory. Relative to a neutral baseline, in which no shapes matched the memory item, search was more efficient when the memory cue matched the shape containing the target, and it was less efficient when the cued stimulus contained a distractor. Cuing affected the shortest reaction times and the first saccade in search. The effect occurred even when the memory cue was always invalid but not when the cue did not have to be held in memory. There was also no evidence for priming effects between consecutive trials. The results suggest that there can be early, involuntary top-down directing of attention to a stimulus matching the contents of working memory. PMID- 15826229 TI - Effector-dependent learning by observation of a finger movement sequence. AB - Can observational learning be effector dependent? In 3 experiments, observers watched a model respond to a 6-item unique sequence in a serial reaction time task. Their sequence knowledge was then compared with that of controls who had performed an unrelated task or observed a model responding to random targets. Observational learning was indicated when the introduction of a new sequence was associated with more reaction time elevation in observers than in controls. The authors found evidence of observational learning only when observers used the finger movement sequence that they observed during training, not when they responded at the same sequence of locations using different digits. Free generation and recognition tests also detected observational learning. These results imply that observational learning can be both explicit and effector dependent. PMID- 15826230 TI - Timing during interruptions in timing. AB - Duration and location of breaks in time interval production were manipulated in various conditions of stimulus presentation (Experiments 1-4). Produced intervals shortened and then stabilized as break duration lengthened, suggesting that participants used the break as a preparatory period to restart timing as quickly as possible at the end of the break. This interpretation was supported in Experiment 5, in which similar results were obtained with a reaction time response executed at the end of the break. In all experiments, produced intervals lengthened as the break occurred later during the interval. The authors conclude that varying break location and duration reveal, respectively, the influence of attentional time-sharing before the interruption and of preparatory processes taking place during the break. PMID- 15826231 TI - Practice increases the efficiency of evidence accumulation in perceptual choice. AB - Most models of choice response time base decisions on evidence accumulated over time. A fundamental distinction among these models concerns whether each piece of evidence is equally weighted (lossless accumulation) or unequally weighted (leaky accumulation). The authors tested a hypothesis derived from A. Heathcote and S. Brown's (2002) self-exciting expert competitor (SEEXC) model of skill acquisition: that evidence accumulation becomes less leaky with practice. The hypothesis was supported by observation that the effects of prime stimuli increased with practice. The authors used metacontrast masked primes, which could not be consciously discriminated by most participants, to avoid methodological problems associated with conscious strategy changes. The form of the law of practice in the data is also shown to be consistent with the SEEXC model. PMID- 15826232 TI - On the limits of advance preparation for a task switch: do people prepare all the task some of the time or some of the task all the time? AB - This study investigated the nature of advance preparation for a task switch, testing 2 key assumptions of R. De Jong's (2000) failure-to-engage theory: (a) Task-switch preparation is all-or-none, and (b) preparation failures stem from nonutilization of available control capabilities. In 3 experiments, switch costs varied dramatically across individual stimulus-response (S-R) pairs of the tasks virtually absent for 1 pair but large for others. These findings indicate that, across trials, task preparation was not all-or-none but, rather, consistently partial (full preparation for some S-R pairs but not others). In other words, people do not prepare all of the task some of the time, they prepare some of the task all of the time. Experiments 2 and 3 produced substantial switch costs even though time deadlines provided strong incentives for optimal advance preparation. Thus, there was no evidence that people have a latent capability to fully prepare for a task switch. PMID- 15826233 TI - The impact of order incongruence between a task-irrelevant auditory sequence and a task-relevant visual sequence. AB - A novel effect is reported in which serial recall of visual digits was disrupted to a greater degree by the presence of the same set of digits presented as an irrelevant auditory sequence than by the presence of irrelevant auditory consonants, but only when the order of the irrelevant digits was incongruent with that of the to-be-remembered digits (Experiment 1). Experiment 2 replicated this order-incongruence effect and showed also that disruption was dictated by the number of order-incongruent transitions but not by the number of novel tokens contained within the irrelevant sequence. The results favor an interference-by- process approach to the disruption of serial memory by irrelevant sound over approaches based on notions of interference by content and/or interference by depletion of attentional resources. PMID- 15826234 TI - Skeletal structure of printed words: evidence from the stroop task. AB - Do readers encode the sequencing of consonant (C) and vowel (V) phonemes (skeletal structure) in printed words? The authors used the Stroop task to examine readers' sensitivity to skeletal structure. In Experiment 1, CVC nonwords (e.g., pof) facilitated the naming of colors with congruent frames (e.g., red, a CVC word) but not with incongruent ones (e.g., green). In Experiment 2, the color black (a CCVC frame) was named faster with a congruent CCVC frame (e.g., grof) compared to either CCVCC (e.g., groft) or CVC (e.g., gof) incongruent controls. Finally, in Experiment 3, the color pink (a CVCC frame) was named faster with a CVCC frame (e.g., goft) compared to either CCVCC or CVC incongruent controls. In most cases, congruent frames shared no segments with the color name. These findings demonstrate that readers automatically assemble the skeletal structure of printed words. PMID- 15826235 TI - Perception of visual speed while moving. AB - During self-motion, the world normally appears stationary. In part, this may be due to reductions in visual motion signals during self-motion. In 8 experiments, the authors used magnitude estimation to characterize changes in visual speed perception as a result of biomechanical self-motion alone (treadmill walking), physical translation alone (passive transport), and both biomechanical self motion and physical translation together (walking). Their results show that each factor alone produces subtractive reductions in visual speed but that subtraction is greatest with both factors together, approximating the sum of the 2 separately. The similarity of results for biomechanical and passive self-motion support H. B. Barlow's (1990) inhibition theory of sensory correlation as a mechanism for implementing H. Wallach's (1987) compensation for self-motion. PMID- 15826236 TI - Visual short-term memory during smooth pursuit eye movements. AB - Visual short-term memory (VSTM) was probed while observers performed smooth pursuit eye movements. Smooth pursuit keeps a moving object stabilized in the fovea. VSTM capacity for position was reduced during smooth pursuit compared with a condition with eye fixation. There was no difference between a condition in which the items were approximately stabilized on the retina because they moved with the pursuit target and a condition in which the items moved across the retina because they were stationary in space. The reduction of capacity for position was eliminated when miniature items were presented on the pursuit target. Similarly, VSTM capacity for color did not differ between smooth pursuit and fixation. The results suggest that visuospatial attention is tied to the target during smooth pursuit, which impairs VSTM for the position of peripheral objects. Sensory memory during smooth pursuit was only slightly impaired. PMID- 15826237 TI - Polymorphism in Nos2 gene is absent in Prague colony of Dahl/Rapp salt-sensitive and salt-resistant rats. AB - We have searched for polymorphism of inducible nitric oxide synthase gene (Nos2 gene) in the Prague colony of salt-sensitive and salt-resistant Dahl/Rapp rats. Specific primers were used to confirm previously described Nos2 gene polymorphism because this gene was suggested to be a potential candidate gene for genetic hypertension. Phenotyping (blood pressure, organ weight, plasma lipids) have confirmed the data known from other colonies of Dahl/Rapp rats. However, in our colony we were not able to find any Nos2 gene polymorphism between salt-sensitive and salt-resistant rats, which was previously described in animals from Harlan colony. Moreover, the genetic homogeneity of our salt-sensitive and salt resistant rats in terms of Nos2 gene was the same as in the original Brookhaven colony of Dahl rats. This is surprising because our colony has been established from breeding pairs kindly provided by Prof. J.P. Rapp more than 15 years ago. It seems that the polymorphism found in Harlan colony could be the result of previous contamination or genetic drift during the breeding conditions specific for this colony. PMID- 15826238 TI - Spontaneous quantal and non-quantal release of acetylcholine at mouse endplate during onset of hypoxia. AB - At 20 (0)C, both quantal and non-quantal spontaneous acetylcholine release (expressed as miniature endplate potential frequency [f-MEPPs] and the H-effect, respectively) increased during the first 30 min of hypoxia in solution with normal extracellular calcium ([Ca(2+)](o) = 2.0 mM). The hypoxia-induced tenfold increase of the f-MEPPs was virtually absent in low calcium solution([Ca(2+)](o) = 0.4 mM) whereas there was still a significant increment of non-quantal release. This indicates that each of these two processes of acetylcholine release is influenced by mechanisms with different oxygen sensitivity. The rise of f-MEPPs during the onset of hypoxia apparently requires Ca(2+) entry into the nerve terminal, whereas the non-quantal release can be increased by another factors such as a lower level of ATP. PMID- 15826239 TI - Secretion of cryptococcal phospholipase B1 (PLB1) is regulated by a glycosylphosphatidylinositol (GPI) anchor. AB - The secreted, multifunctional enzyme PLB1 (phospholipase B1 protein encoded by the PLB1 gene) is a virulence determinant of the pathogenic fungus Cryptococcus neoformans, but the mechanism of its secretion is unknown. The cryptococcal PLB1 gene encodes putative, N-terminal LP (leader peptide) and C-terminal GPI (glycosylphosphatidylinositol) anchor attachment motifs, suggesting that PLB1 is GPI-anchored before secretion. To investigate the role of these motifs in PLB1 secretion, four cDNA constructs were created encoding the full-length construct (PLB1) and three truncated versions without the LP and/or the GPI anchor attachment motifs [(LP-)PLB1 (PLB1 expressed without the LP consensus motif), (LP )PLB1(GPI-) (PLB1 expressed without the LP and GPI consensus motifs) and PLB1(GPI ) (PLB1 expressed without the GPI anchor attachment motif) respectively]. The constructs were ligated into pYES2, and galactose-induced expression was achieved in Saccharomyces cerevisiae. The LP was essential for secretion of the PLB1 protein and its three activities (PLB, lysophospholipase and lysophospholipase transacylase). Deletion of the GPI motif to create PLB1(GPI-) resulted in a redistribution of activity from the cell wall and membranes to the secreted and cytosolic fractions, with 36-54% of the total activity being secreted as compared with <5% for PLB1. PLB1 produced the maximum cell-associated activity (>2-fold more than that for PLB1(GPI-)), with 75-86% of this in the cell-wall fraction, 6 19% in the membrane fraction and 3-7% in the cytosolic fraction. Cell-wall localization was confirmed by release of activity with beta-glucanase in both S. cerevisiae recombinants and wild-type C. neoformans. The dominant location of PLB1 in the cell wall via GPI anchoring may permit immediate release of the enzyme in response to changing environmental conditions and may represent part of a novel mechanism for regulating the secretion of a fungal virulence determinant. PMID- 15826240 TI - The caspase family in urochordates: distinct evolutionary fates in ascidians and larvaceans. AB - BACKGROUND INFORMATION: Caspases are cysteine proteases that mediate apoptosis (programmed cell death) initiation and execution. Apoptosis is a conserved mechanism shared by all metazoans, although its physiological function and complexity show considerable taxon-dependent variations. To gain insight into the caspase repertoire of putative ancestors to vertebrates, we performed exhaustive genomic searches in urochordates, a sister taxon to vertebrates in which ascidians and appendicularians display chordate characters at early stages of their development. RESULTS: We identified the complete caspase families of two ascidians (Ciona intestinalis and C. savignyi) and one larvacean (Oikopleura dioica). We found in ascidian species an extremely high number of caspase genes (17 for C. intestinalis and 22 for C. savignyi), deriving from five founder gene orthologues to human pro-inflammatory, initiator and executioner caspases. Although considered to be sibling species, C. intestinalis and C. savignyi only share 11 orthologues, most of the additional genes resulting from recent mass duplications. A sharply contrasted picture was found in O. dioica, which displayed only three caspase genes deriving from a single founder gene distantly related to caspase 3/7. The difference between ascidian and larvacean caspase repertoires is discussed in the light of their developmental patterns and life cycles. CONCLUSIONS: The identification of caspase members in two ascidian species delineates five founder genes that bridge the gap between vertebrates and Ecdysozoa (arthropods and nematodes). Given the amazing diversity among urochordates, determination and comparison of the caspase repertoires in species from orders additional to Enterogona (ascidians) and Oikopleuridae might be highly informative on the evolution of caspase-dependent physiological processes. PMID- 15826241 TI - The N370S (Asn370-->Ser) mutation affects the capacity of glucosylceramidase to interact with anionic phospholipid-containing membranes and saposin C. AB - The properties of the endolysosomal enzyme GCase (glucosylceramidase), carrying the most prevalent mutation observed in Gaucher patients, namely substitution of an asparagine residue with a serine at amino acid position 370 [N370S (Asn370- >Ser) GCase], were investigated in the present study. We previously demonstrated that Sap (saposin) C, the physiological GCase activator, promotes the association of GCase with anionic phospholipid-containing membranes, reconstituting in this way the enzyme activity. In the present study, we show that, in the presence of Sap C and membranes containing high levels of anionic phospholipids, both normal and N370S GCases are able to associate with the lipid surface and to express their activity. Conversely, when the amount of anionic phospholipids in the membrane is reduced (approximately 20% of total lipids), Sap C is still able to promote binding and activation of the normal enzyme, but not of N370S GCase. The altered interaction of the mutated enzyme with anionic phospholipid-containing membranes and Sap C was further demonstrated in Gaucher fibroblasts by confocal microscopy, which revealed poor co-localization of N370S GCase with Sap C and lysobisphosphatidic acid, the most abundant anionic phospholipid in endolysosomes. Moreover, we found that N370S Gaucher fibroblasts accumulate endolysosomal free cholesterol, a lipid that might further interfere with the interaction of the enzyme with Sap C and lysobisphosphatidic acid-containing membranes. In summary, our results show that the N370S mutation primarily affects the interaction of GCase with its physiological activators, namely Sap C and anionic phospholipid-containing membranes. We thus propose that the poor contact between N370S GCase and its activators may be responsible for the low activity of the mutant enzyme in vivo. PMID- 15826242 TI - CD109 expression in squamous cell carcinoma of the uterine cervix. AB - CD109 is a cell surface protein, a member of the alpha(2) macroglobulin/C3,C4,C5 family of thioester-containing proteins. The authors have recently reported that high expression of the CD109 gene was detected in approximately half of the examined lung and esophageal squamous cell carcinomas as well as in the testis, and that CD109 has the characteristics of a cancer-testis antigen. In the present study CD109 expression in cervical squamous cell carcinoma was compared with that in endometrial adenocarcinoma by reverse transcription polymerase chain reaction (RT-PCR). The result demonstrated that CD109 expression is significantly higher in cervical squamous cell carcinomas than in endometrial adenocarcinomas and normal cervix and endometrium. In contrast, when expression of RET finger protein (RFP) and bromodomain testis-specific (BRDT) genes, which are also known to be highly expressed in the testis, was examined, no significant difference in their expression levels was observed between squamous cell carcinomas and adenocarcinomas. These findings suggest that CD109 may become a molecular target for the development of new therapeutics for squamous cell carcinoma of various tissue origins. PMID- 15826243 TI - Coexistence of gastric- and intestinal-type endocrine cells in gastric and intestinal mixed intestinal metaplasia of the human stomach. AB - Intestinal metaplasia (IM) in the human stomach has previously been classified into a gastric and intestinal mixed (GI-IM) and a solely intestinal phenotype (I IM). The phenotypes of mucous and endocrine cells were evaluated in 3034 glandular ducts associated with chronic gastritis. In the pyloric region, the relative expression of gastric endocrine cell markers, such as gastrin and somatostatin, decreased gradually from glandular ducts with only gastric mucous cell phenotype (G type) to GI-IM toward I-IM, while that of the intestinal endocrine cell markers, glicentin, gastric inhibitory polypeptide (GIP), and glucagon-like peptide-1 (GLP-1) was inversely correlated. In the fundic region, gastrin-positive cells emerged in the pseudo-pyloric and GI-IM glands, whereas I IM glands did not possess any gastrin-positive cells, suggesting the presence of a distinct pathway of intestinalization. Double staining revealed coexistence of gastrin- and GLP-1-positive cells in the same gland and occasionally in the same cell in GI-IM glands. These results suggest that the phenotypes of endocrine cells are in line with those for mucous counterparts and support the concept that all of the different types of mucous and endocrine cells in normal and IM glands might be derived from a single progenitor cell in each gland. PMID- 15826244 TI - Proposal of histological criteria for intraepithelial atypical/proliferative biliary epithelial lesions of the bile duct in hepatolithiasis with respect to cholangiocarcinoma: preliminary report based on interobserver agreement. AB - Biliary lining epithelia of the bile ducts in biliary diseases are known to have intraepithelial atypical/proliferative lesions related to the development of cholangiocarcinoma. The purpose of the present study was to determine the histological criteria for these lesions based on interobserver agreement. Digital images of 30 intraepithelial atypical/proliferative lesions in the stone containing intrahepatic bile ducts of hepatolithiasis (30 cases) were sent to 10 pathologists. At first, 10 pathologists made a diagnosis (either of reactive/regenerative change, low-grade or high-grade biliary intraepithelial neoplasia (BilIN-1 and BilIN-2), or in situ carcinoma (BilIN-3)) based on their own criteria. The histological criteria for these four lesions were then determined, and the digital images of the same lesions with proposed criteria were re-distributed. Interobserver agreement on these four lesions was slightly improved (kappa = 0.44, first diagnosis; 0.49, second diagnosis) and intraobserver agreement was 'almost perfect' (kappa = 0.82 at both first and second diagnosis). Interobserver agreement between BilIN-1 and BilIN-2 and that between BilIN-2 and BilIN-3 were 'moderate', although the agreement between regenerative/reactive change and BilIN-1 was 'fair'. In this report, we propose histological criteria for reactive/regenerative change, BilIN-1, BilIN-2 and BilIN-3. Improvement of interobserver agreement suggests their applicability in diagnostic and research fields. PMID- 15826245 TI - Post-inflammatory glomerular remodeling is influenced by transformed mesangial cells. AB - To identify the role of transformed mesangial cells (MC) during glomerular remodeling, anti-thymocyte-1 (Thy1) nephritis; modified Thy1 nephritis (injections of anti-Thy1 antibody four times, weekly); and Thy1 nephritis treated with signal transduction inhibitor 571 (Thy1 + STI); were analyzed. At week 1 the index of MC proliferation in modified Thy1 nephritis and in mesangiolysis in Thy1 + STI nephritis was highest among the three models. From week 4, the index of alpha-smooth muscle actin (alpha-SMA) was significantly higher in modified Thy1 nephritis than the other two models. Production of the mesangial matrix including type IV collagen was increased in modified Thy1 but inhibited in Thy1 + STI nephritis. In contrast to modified Thy1 nephritis, the capillary numbers in glomeruli recovered to normal at week 4 in Thy1, and at week 8 in Thy1 + STI nephritis. At week 12, both the adhesive and sclerotic index was significantly higher in modified Thy1 than in the other two models. Data suggest that a moderate amount of mesangial matrix results in a complete repair of capillary loops. Overproduction of the mesangial matrix retards capillary remodeling and finally induces glomerulosclerosis. Insufficient mesangial matrix delays the repair of capillary loops. In conclusion, transformed MC may influence glomerular remodeling by changing the amount of mesangial matrix. PMID- 15826246 TI - Zygomycosis involving lungs, heart and brain, superimposed on pulmonary edema. AB - Zygomycosis is an uncommon but frequently fatal infection and occurs mostly in immunosuppressed hosts, whereas approximately 50% of zygomycosis occurs in diabetic patients. The current patient initially presented with persistent pulmonary edema secondary to renal failure. This was the last of four admissions within 1 year for this 68-year-old woman, for whom the chief complaints were shortness of breath and chest pain. Her past medical history included insulin requiring type 2 diabetes and hypertension for 10 years, and chronic heart and renal failure. She was previously admitted to the hospital for what appeared to be pulmonary edema secondary to renal failure. In the last admission the patient developed pulmonary hemorrhage and metabolic acidosis. Transbronchial biopsy was performed, showing irregular fungal hyphae in the blood vessels, morphologically consistent with zygomycosis. Central nervous system computed tomography also revealed a large infarct in the cerebral hemisphere. The patient died on the seventh hospital day. At autopsy three organs were extensively involved by zygomycosis: (i) lungs were diffusely hemorrhagic with acute infarcts; (ii) pericardium had fibrotic inflammation; and (iii) the left cerebral hemisphere, cerebellum and pons had large hemorrhagic infarct by zygomycosis infection. Corticosteroid medication and hemodialysis triggered increasing hyperglycemia, metabolic acidosis and iron overload, which contributed to zygomycosis infection that subsequently spread to the heart and brain as a rare consequence. PMID- 15826247 TI - Intrahepatic cholangiocarcinoma with multicystic, mucinous appearance and oncocytic change. AB - A case is reported herein of intrahepatic cholangiocarcinoma (ICC) with multicystic, mucinous appearance and oncocytic change. Because of liver dysfunction, a 73-year-old woman was hospitalized in early October 2003. She was diagnosed as having ICC of the right hepatic lobe with occlusion of the hilar and perihilar bile ducts by imaging examination. Extended right lobectomy was performed but the patient died of liver failure on the next day. In surgically resected specimens, the tumor (3 x 3 cm) was mainly located in the right lobe, and tumors infiltrated along the biliary tree as well as invading into the adjacent hepatic parenchyma. The tumor mass had a sponge or honeycomb appearance. Microscopically, these tumors were composed of multiple microcysts filled by abundant mucin and lined by micropapillary adenocarcinoma cells. Their cytoplasm was acidophilic, appearing as an oncocyte, and carcinoma cells were positive for mitochondrial antigen in addition to biliary cytokeratins. There were no ovarian like stromas around these cystic tumors, and communication of the biliary lumen with these carcinomatous cysts was not evident, thus different from biliary mucinous cystadenocarcinoma and intraductal papillary neoplasm of the liver. This is the third case of multicystic mucinous ICC and the present case might have been derived from intrahepatic peribiliary glands. PMID- 15826248 TI - Renal oncocytosis. AB - Renal oncocytosis is a rare disorder in which numerous oncocytic nodules develop in the kidney. An additional case is reported here. The patient was a 51-year-old woman who had received hemodialysis for 27 years. Nineteen years previously she had developed a tumorous lesion in the right kidney, which had been diagnosed as oncocytoma with laparotomic biopsy. Recently the kidney was removed because of enlargement of the tumor. The renal parenchyma was entirely replaced with numerous brownish nodules. Histologically, the nodules were composed of nests of uniform oncocytic cells. Ultrastructurally, the oncocytic cells contained numerous mitochondria. Immunohistochemical features of the nodules were identical to those of sporadic oncocytomas, that is, immunophenotypes similar to the distal nephron and reactivity with antimitochondrial antigen. Based on these findings, the lesion was diagnosed as renal oncocytosis. It was not possible to determine whether the larger nodules should be diagnosed as oncocytoma or a part of oncocytosis. Additionally, the germ line mutation of the Birt-Hogg-Dube (BHD) syndrome gene was examined using the genomic DNA obtained from the peripheral lymphocytes, which failed to show any gene alteration. Despite the rare occurrence pathologists and urologists should be aware of renal oncocytosis, as a precursor lesion of renal oncocytoma and chromophobe renal cell carcinoma. PMID- 15826249 TI - Autopsy case of primary choriocarcinoma of the urinary bladder. AB - Choriocarcinomas usually develop in the uterus and ovaries in the female, being extremely rare in the extragenital organs in the male. Extragenital choriocarcinomas in the male usually develop in the mediastinum or retroperitoneum. The frequency of choriocarcinoma in the urinary bladder is extremely low. The purpose of the present paper was to report an autopsy case of choriocarcinoma in the urinary bladder in the male. An 81-year-old male patient with macrohematuria was first diagnosed with transitional cell carcinoma (TCC). At autopsy a hemorrhagic necrotic tumor, which was found in the urinary bladder with metastatic lesions in the lungs, was diagnosed as choriocarcinoma microscopically. There was no evidence for choriocarcinoma derived from any other organs than the urinary bladder, although there were metastatic lesions in both lungs and the direct invasion into the prostate. From these findings it is concluded that the tumor was a primary choriocarcinoma in the urinary bladder in a male patient. Choriocarcinoma of the urinary bladder is very rare, but the prognosis is extremely poor in comparison with TCC even in the urinary bladder. Therefore, it is essential to clearly discriminate between choriocarcinomas and TCC. PMID- 15826252 TI - MRI: safety in patients with pacemakers or defibrillators: is it prime time yet? PMID- 15826250 TI - Amyloidosis of pancreatic islets in primary amyloidosis (AL type). AB - Seven cases of primary amyloidosis (AL-type) were studied immunocytochemically for the possible involvement of pancreatic islets. The two cases with extensive organ involvement by AL-amyloidosis revealed amyloid deposits in pancreatic islets by routine HE and Congo red staining, which were positive for amyloid p and amyloid a, but were only focally positive for light chains kappa and lambda. Positive staining for amyloid p and amyloid a was also noted in the scattered pancreatic acinar tissues, and this positive staining was not specifically located in pancreatic islets as seen in type 2 diabetes mellitus. It is concluded that amyloid deposits in pancreatic islets occur in systemic AL-amyloidosis by a different mechanism from type 2 diabetes. Islet amyloidosis in AL-amyloidosis appears to deposit via circulation, depositing in both pancreatic islets and acinar tissue through blood vessels. In type 2 diabetes, beta islet cells die by cytotoxic effects of smaller amylin (islet amyloid polypeptide, IAPP) aggregates, and the interstitial space created by the necrotic beta cells is replaced by larger IAPP aggregates, to form complex, polymerized islet amyloid. In AL amyloidosis, the amount of amyloid and light chain deposits in pancreatic islets is much less than that of the other organs and appears to have no connection to type 2 diabetes because the patients did not present diabetes or hyperglycemia. However, considerable islet amyloidosis can be seen in severe AL-type amyloidosis. PMID- 15826253 TI - Industry viewpoint: Guidant: Pacemakers, ICDs, and MRI. PMID- 15826254 TI - Industry viewpoint: Medtronic: Pacemakers, ICDs, and MRI. PMID- 15826255 TI - Industry viewpoint: St. Jude Medical: Pacemakers, ICDs and MRI. PMID- 15826256 TI - Government viewpoint: U.S. Food & Drug Administration: Pacemakers, ICDs and MRI. PMID- 15826257 TI - Outcome of magnetic resonance imaging (MRI) in selected patients with implantable cardioverter defibrillators (ICDs). AB - OBJECTIVE: To determine if simple strategies used to safely scan pacemaker patients could be applied to implantable cardioverter defibrillator (ICD), patients undergoing MRI allowing ICD patients to undergo MRI as well. INTERVENTIONS: Screening, reprogramming, and monitoring strategies were used to facilitate MRI. RESULTS: Seven patients underwent eight MRI scans at 1.5 T. Post MRI, all devices demonstrated no change in pacing, sensing, impedances, charge times, or battery status. The patient undergoing a lumbar spine scan experienced a "power-on-reset" of his ICD without permanent impairment of his device. CONCLUSION: Scanning of ICD patients might be performed if appropriate reprogramming and monitoring is implemented. PMID- 15826258 TI - Is magnetic resonance imaging safe in cardiac pacemaker recipients? AB - Magnetic resonance imaging (MRI) is currently contraindicated in cardiac pacemaker (PM) recipients. The objectives of this prospective study were to (1) reassess the risks of performing an MRI scan in patients with PM, (2) compared the pacing functions before and after the exposure to MRI, and (3) monitor the development of possible adverse effects. Thirteen patients implanted with an Affinity DR model 5330 PMs (St. Jude Medical) connected to a Tendril model 1388 leads (St. Jude Medical) underwent 2.0 T-MRI for a variety of indications. All patients displayed a stable spontaneous rhythm at the time of the MRI scan and were not considered to be PM-dependent. The sensing and pacing functions were analyzed and the impedance of both leads was measured before and after the scan. The MRI scan was performed with all PM programmed in DDD mode. The sensing configuration was bipolar. All patients were monitored utilizing a standard electrocardiographic monitor and direct verbal communication. PM Inhibition, asynchronous pacing, or inappropriately rapid pacing was not observed. No patient reported discomfort, heat, or motion sensation at the PM implant site. There were no significant differences in the sensing, stimulation, AutoCapture threshold, and lead impedance measurements before and after MRI. The results of this study suggest that performing 2.0 T-MRI scans in patients with Affinity DR model 5330 PM connected to a Tendril model 1388 lead is safe. PMID- 15826259 TI - Rate-dependent AV delay optimization in cardiac resynchronization therapy. AB - BACKGROUND: During cardiac resynchronization therapy (CRT), cardiac performance is dependent on an optimized atrioventricular delay (AVD). However, the optimal AVD at different heart rates has not been defined yet during CRT. METHOD: The effects of an increase in heart rate by pacing or physical exercise on optimal AVD were studied in 36 patients with biventricular pacemakers/defibrillators. The velocity time integral (VTI) in the left ventricular outflow tract (LVOT) was measured with pulsed Doppler either at three different paced heart rates in the supine position or in seated position before and after physical exercise. RESULTS: The baseline AVD was optimized to 99 +/- 19 ms in the supine and 84 +/- 22 ms in the seated position. When the heart rate was increased by DDD pacing, there was a positive linear relationship between an increase in heart rate, in AVD and in VTI (LVOT-VTI + 0.047 cm/s per 10 beats per minute (bpm) heart rate increase per 20 ms increase in AVD, P = 0.007). A similar but more pronounced relationship was found after physical exercise in the seated position (LVOT-VTI + 0.146 cm/s per 10 bpm heart rate increase per 20 ms increase of AVD, P = 0.013). This effect was observed in patients with and without AV block and mitral regurgitation. CONCLUSIONS: In conclusion, the systolic performance of the dilated ventricle, which depends on an elevated preload, is critically affected by the appropriate timing of the AVD during exercise. In contrast to normal pacemaker patients, in CRT the relatively short baseline AVD should be prolonged at increased heart rates. Further studies with other means of measuring exercise cardiac performance are needed to confirm these unexpected findings. PMID- 15826261 TI - Atrial pacing should be used more frequently in sinus node disease. AB - BACKGROUND: Single lead atrial (AAI(R)) pacing for sick sinus syndrome provides physiological pacing and is less expensive than a dual chamber system. Compared with ventricular-based pacing, it maintains the normal cardiac depolarization sequence, is associated with less atrial fibrillation and heart failure, and avoids pacemaker syndrome. We sought whether it is possible to select patients for AAI(R) pacing with a low likelihood of subsequent ventricular pacing, and whether this approach was cost-effective. METHODS: A retrospective review was conducted of AAI(R) pacemaker implantations. Patients requiring a further procedure for insertion of a ventricular lead (for DDD(R) or VVI(R) pacing) were compared with those who remained atrially paced. RESULTS: 2.7% (117 of 4,366) of implants were AAI(R), compared with less than 1% overall for New Zealand and Australia. During follow-up of 3.5 (2.3, 7.7) years, insertion of a ventricular lead was required in 14 (12%), and was more likely in those with pre-existing PR interval >0.20 seconds (odds ratio 7.8, P = 0.003) or left bundle branch block (LBBB, odds ratio 17, P = 0.037). Right bundle branch block, left anterior fascicular block, or history of paroxysmal atrial fibrillation were not more frequent in the group requiring ventricular pacing, and preimplantation Wenckebach point was not different. The most efficient strategy was initial AAIR implantation in all except those with LBBB or PR interval >0.20 seconds. Compared with routine DDDR implantation, cost was reduced by 20%, with a 1.4% annual incidence of further procedures. CONCLUSION: AAIR is the appropriate pacing choice for sick sinus syndrome without LBBB or PR interval >0.20 seconds. PMID- 15826260 TI - Electrophysiologic study-guided therapy with sotalol for life-threatening ventricular tachyarrhythmias. AB - The aim of this study was to investigate the long-term efficacy and safety of electrophysiologic study (EPS)-guided sotalol administration combined with implantable cardioverter defibrillators (ICD) for ventricular tachyarrhythmias (VTA). This study enrolled 92 patients with both structural heart disease and sustained VTA. Sotalol was administered to 57 patients, and its efficacy was assessed by EPS. Long-term treatment was continued in combination with ICD in 31 patients (57%) whose VTA was no longer inducible (responder group) and in 16 patients whose VTA remained inducible (nonresponder group). The long-term outcomes were compared among the responder group, the nonresponder group, and 35 ICD recipients untreated with antiarrhythmic drugs (ICD-only group). During a mean follow-up of 44 +/- 33 months, the recurrence of VTA was not significantly different between all patients treated with sotalol (30%) and patients in the ICD only group (46%). However, the recurrence of VTA was significantly lower in the responder (13%) than in the nonresponder (63%) or the ICD-only groups (46%). There was no significant difference in VTA recurrence between the nonresponder and the ICD-only groups. One patient each in the responder and the ICD-only groups died suddenly, and all-cause mortality was similar in the three groups. The incidence of inappropriate ICD discharges was less in the sotalol than in the ICD-only groups. No patient had to discontinue long-term sotalol treatment because of the adverse effects. In conclusion, sotalol reduced VTA recurrence in the responding patients and inappropriate ICD discharge. EPS may predict the efficacy of sotalol for VTA recurrence. PMID- 15826262 TI - Transcoronary ablation of septal hypertrophy does not alter ICD intervention rates in high risk patients with hypertrophic obstructive cardiomyopathy. AB - INTRODUCTION: Transcoronary ablation of septal hypertrophy (TASH) is safe and effectively reduces the intraventricular gradient in patients with hypertrophic obstructive cardiomyopathy (HOCM). To analyze the potential of anti- and proarrhythmic effects of TASH, we studied the discharge rates of implanted cardioverter defibrillators (ICD) in patients with HOCM who are at a high risk for sudden cardiac death. METHODS: ICD and TASH were performed in 15 patients. Indications for ICD-implantation were secondary prevention in nine patients after resuscitation from cardiac arrest with documented ventricular fibrillation (n = 7) or sustained ventricular tachycardia (n = 2) and primary prevention in 6 patients with a family history of sudden deaths, nonsustained ventricular tachycardia, and/or syncope. All the patients had severe symptoms due to HOCM (NYHA functional class = 2.9). RESULTS: During a mean follow-up time of 41 +/- 22.7 months following the TASH procedure, 4 patients had episodes of appropriate discharges (8% per year). The discharge rate in the secondary prevention group was 10% per year and 5% in the group with primary prophylactic implants. Three patients died during follow-up (one each of pulmonary embolism, stroke, and sudden death). CONCLUSION: In conclusion, on the basis of ICD-discharge rates in HOCM-patients at high risk for sudden death, there is no evidence for an unfavorable arrhythmogenic effect of TASH. The efficacy of ICD treatment for the prevention of sudden cardiac death in HOCM could be confirmed, however, mortality is high in this cohort of hypertrophic cardiomyopathy patients. PMID- 15826263 TI - Editorial comment: Operation and failure. PMID- 15826264 TI - Cardiac resynchronization induces favorable neurohumoral changes. AB - AIM: The aim of this article is to examine whether cardiac resynchronization therapy (CRT) induces improvements in the neurohumoral system. METHODS AND RESULTS: Thirteen patients with HF (left ventricular (LV) ejection fraction <35%) were included. Before and after 6 months of CRT, myocardial (123)I metaiodobenzylguanidine ((123)I-MIBG) uptake indices, used as an index of neural norepinephrine reuptake and retention, and brain natriuretic peptide (BNP) levels, used as an index of LV end-diastolic pressure, NYHA classification and echocardiographic indices were assessed. Six months of CRT resulted in significant improvement in (1) NYHA classification and reduction in QRS width (P < 0.001), (2) decrease of LV end-diastolic diameter (P = 0.005), LV end-systolic diameter (P = 0.005), septal to lateral delay (P = 0.01) and mitral regurgitation (MR, P = 0.04), (3) delayed (123)I-MIBG heart/mediastinum ratios improved (P = 0.03) and (123)I-MIBG washout decreased (P = 0.001), and (4) BNP levels decreased (P = 0.001). CONCLUSIONS: Parallel to significant functional improvement and echocardiographic reverse remodeling and resynchronization, our data indicate that CRT induces favorable changes in the neurohumoral system. PMID- 15826265 TI - A comparative study of the action of dexamethasone sodium phosphate and dexamethasone acetate in steroid-eluting pacemaker leads. AB - BACKGROUND: The aim of this study was to characterize acute and medium-term pacemaker lead performance with the two most commonly used glucocorticosteroids: dexamethasone sodium phosphate and dexamethasone acetate. METHODS: Forty sets of atrial and ventricular passive-fixation leads containing either dexamethasone sodium phosphate or dexamethasone acetate were implanted as dual chamber pacemakers. Randomization was equally distributed to both arms of the study. Stimulation thresholds, lead impedance, and sensing were measured on the day of implant, day 1, 1 month, 3 months, and 6 months following the implant. RESULTS: For the dexamethasone sodium phosphate arm, the atrial stimulation thresholds were 0.9 +/- 0.1 V at implant and 0.8 +/- 0.1 V at 6 months, and in the ventricle 0.5 +/- 0.1 V at implant and 0.6 +/- 0.1 V at 6 months. In the dexamethasone acetate arm, the atrial stimulation thresholds were 0.7 +/- 0.1 V at implant and at 6 months, and in the ventricle 0.5 +/- 0.1 V at implant and at 6 months. There were no significant differences between dexamethasone sodium phosphate or dexamethasone acetate leads for stimulation thresholds at any of the intervals of follow-up. P- and R-wave sensing were similarly maintained over the duration of follow-up with no significant differences between groups at any of the intervals of follow-up. Pacing lead impedance showed a trend towards lower values in the dexamethasone acetate arm, which only reached statistical significance at 3 months and beyond for ventricular leads. CONCLUSIONS: Leads containing dexamethasone sodium phosphate and dexamethasone acetate demonstrate equivalent and excellent acute and medium-term pacemaker lead performance characteristics. PMID- 15826266 TI - Use of advanced mapping systems to guide ablation in complex cases: experience with noncontact mapping and electroanatomic mapping systems. AB - OBJECTIVE: This report describes our experience with noncontact mapping and electroanatomic mapping in complex ablations, which are defined as ablations done after failure of conventional ablation. MATERIAL AND METHODS: Patients were included (N = 68; 49% with structural heart disease) in whom previous ablation failed and in whom a second procedure was done with advanced mapping. Non-contact mapping was used in 17 patients, electroanatomic mapping in 36, and both noncontact and electroanatomic mapping in 15. Arrhythmias included focal atrial tachycardia (n = 16), reentrant atrial tachycardia (n = 14), right ventricular outflow tachycardia (n = 10), post-myocardial infarction ventricular tachycardia (n = 9), and others (n = 19). RESULTS: Acute success at the second ablation was achieved in 79% of patients. At 20 +/- 9 months after the procedure, 69% of these patients reported having significantly fewer symptoms than before the second ablation, and 51% were free of symptoms. Only 16% were using antiarrhythmic medications. Complications included a small pericardial effusion in two patients, hypotension in one patient, and a femoral pseudoaneurysm in another. CONCLUSIONS: Advanced mapping is a useful and safe adjunct for catheter ablation after ablation has failed in patients with complex substrate. PMID- 15826267 TI - Introduction of permanent cardiac stimulation/defibrillation leads via the retro pectoral veins. AB - When the cephalic vein route is not easily accessible for the introduction of permanent stimulation/defibrillation leads, retro-pectoral veins can be looked for, which are usually present and suitable in most patients. As with the cephalic vein route, it is a safer approach than direct subclavian vein puncture. Moreover, using a guidewire and a split introducer increases the rate of successful cannulation. PMID- 15826268 TI - Current clinical issues for MRI scanning of pacemaker and defibrillator patients. AB - Dramatic increases in both magnetic resonance imaging (MRI) usage and cardiac device-based therapy have resulted in an estimated 50-75% probability of a patient being indicated for an MRI over the lifetime of their device. Some recent studies have demonstrated "safe procedures" and "no adverse events" in the limited populations, clinical situations, and specific devices and lead orientations tested. While these investigations are useful to help ascertain the hazards for patients with cardiac devices, they do not demonstrate clear freedom from risk. All components of active implantable systems must be engineered during the design stage to provide safety in current and evolving MR environments. Device manufacturers need to secure regulatory approval to confirm their products' safety under multiple clinical and technical variables. PMID- 15826269 TI - The enigma of statistics and modern cardiology. PMID- 15826271 TI - Phantom crosstalk. PMID- 15826272 TI - Cardiac magnetic resonance imaging in a patient with implantable cardioverter defibrillator. AB - The presence of pacemakers and implantable cardioverter-defibrillators (ICD) is considered historically a contraindication to magnetic resonance (MR) imaging. This image modality has unparalleled soft-tissue imaging capabilities, and many consider it as the image of choice for patients with arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C). ICDs are now smaller, with less magnetic materials and improved electromagnetic interference protection. We tested modern ICDs for heat, force, function and image distortion and found that several of them may indeed be MRI safe. We report here a patient who was suspected of ARVD/C, underwent ICD implantation based on MR safety testing, and underwent intentionally scheduled follow-up cardiac MR imaging. This is the description of a patient with an ICD who had planned MRI scanning. The scan was safe and most of the MRI images were of high quality. PMID- 15826273 TI - Safe performance of magnetic resonance imaging on a patient with an ICD. AB - This is a report on a patient with an implanted cardioverter defibrillator (ICD) who intentionally underwent magnetic resonance imaging (MRI) of a malignant brain tumor. To avoid inadequate detection of ventricular tachycardia (VT) or ventricular fibrillation (VF), the ICD was inactivated by programming the VT detection and VT/VF-therapy status off. The patient came through the protocol safely and without any difficulty or discomfort. There was no arrhythmic event. MRI affected neither programmed data nor the function of the ICD system. PMID- 15826274 TI - Pleomorphic ventricular tachycardia: an uncommon presentation in idiopathic left ventricular tachycardia. AB - Idiopathic left ventricular tachycardia (ILVT) is a distinct entity that arises in the left ventricle, may have reentrant mechanism and is verapamil-sensitive. Pleomorphism as defined by multiple ventricular tachycardia morphologies is usually associated with either coronary artery disease or cardiomyopathy but very rare in cases of ILVT. In this case report, we describe an unusual case of ILVT with two ECG morphologies of the opposite axis that were successfully eliminated with radiofrequency ablation. The successful ablation sites were closely located to each other in the left lower ventricular septum. PMID- 15826275 TI - Infrequent syncope in a heavy vehicle driver. AB - We present a case study of a truck driver with sinus node dysfunction diagnosed with the help of an implantable loop recorder more than 18 months after implantation. This case emphasizes that cardiogenic syncope may be very infrequent and its diagnosis can be elusive despite extensive testing. PMID- 15826276 TI - Isotretinoin (13-cis-retinoic acid) associated atrial tachycardia. AB - We describe a 16-year-old boy who presented with palpitations for 1 week while being on isotretinoin treatment for nodulocystic facial acne for 3 months. Twenty four-hour Holter monitoring showed frequent premature atrial beats and episodes of nonsustained atrial tachycardia. He never had any episodes of palpitations previously. His complaints almost disappeared within a week after stopping the treatment. He remained asymptomatic since the discontinuation of the drug. The temporal relationship between isotretinoin treatment and patient's symptoms in the presence of documented arrhythmia suggests a drug-related cause. As a result, clinicians should be aware of the possible arrhythmogenic effect of isotretinoin. PMID- 15826277 TI - Late perforation of the heart, pericardium, and diaphragm by an active-fixation ventricular lead. AB - This case report describes a late perforation of right heart wall, pericardium, and diaphragm by an active-fixation ventricular lead without development of pericardial effusion and cardiac tamponade even under oral warfarin treatment. PMID- 15826278 TI - Longitudinally partitioned coronary sinus: an unusual anomaly of the coronary venous system. AB - We report a case of a patient with advanced heart failure who had a longitudinally partitioned coronary sinus. With multidirectional fluoroscopic views and a careful approach to the target lumen, a left ventricular lead for biventricular pacing was placed successfully in the left marginal vein. PMID- 15826279 TI - Re.: Complete loss of ICD programmability after magnetic resonance imaging. PMID- 15826281 TI - Clinical evaluation of Aspergillus-PCR for detection of invasive aspergillosis in immunosuppressed patients. AB - We evaluated the utility of Aspergillus PCR as a tool in diagnosing invasive aspergillosis in patients at risk. Aspergillosis was assessed according to the European Organization for Research and Treatment of Cancer (EORTC), Mycosis Study Group definitions. Nine and seven patients with proven and probable aspergillosis respectively were evaluated. Whole blood samples prior (n = 41) and during antifungal treatment (n = 67), and tissue specimens (n = 9) and/or bronchoalveolar lavage fluids (n = 7) were investigated. In patients with proven infections, the sensitivities of PCR of lung samples were 100%, of blood samples prior treatment were 66%, and during treatment 55%. Clearance of fungal DNA from blood was associated with resolution of clinical symptoms in two of four patients with proven infection. Consecutive positive PCR results for Aspergillus are fatal as two of five patients died. In patients with probable infections, the sensitivities of PCR of lung fluids were 85%, of blood samples prior treatment were 57%, and during treatment 42%. The benefits of PCR diagnosing and screening of whole blood are limited if sampling takes place once treatment has started. The performance of Aspergillus PCR from tissue samples should be recommended in addition to microscopic examination and culture technique for sensitive detection of fungal infection. PMID- 15826282 TI - [Fungi in the mediastinum: rare, but relevant]. AB - Fungal infections of the mediastinum are rare. There are five forms to be differentiated: (i) hematogenous fungal infection of the mediastinum after fungemia; (ii) postoperative fungal infection after cardiosurgical sternotomy; (iii) as special form of a descending necrotizing fasciitis; (iv) infection of the mediastinum per continuitatem; (v) fungal infection of the posterior mediastinum after esophageal perforation. In the surgical department, University of Schleswig-Holstein, Campus Lubeck, five patients with a fungal mediastinitis were treated during a period of 10 years. Three patients suffered from a descending mediastinitis, one patient from a fungal infection after pseudarthrosis of the sternum (postoperatively after cardiosurgery). The section results of one other patient with pulmonary aspergillosis showed a complete invasion of the mediastinum by Aspergillus fumigatus. PMID- 15826283 TI - [Intra-abdominal mycoses]. AB - The incidence of invasive mycoses in patients undergoing abdominal surgery amounts to approximately 8% and shows an upward trend in epidemiological studies. The lethality of these systemic mycoses, which are mostly based on Candida infections constitutes up to 60%. The development of a sytemic mycosis is marked by exogenic, endogenic and iatrogenic risk factors and typically displays tissue invasion after an initial fungal contamination or systemic dissemination via fungal sepsis. Fungal peritonitis is generally a monoinfection with Candida spp., where Candida albicans outweighs in 70% of cases. Aspergillus spp. are only detected abdominally in rare cases. The histological verification of a fungal invasion is regarded as proof of the existence of an invasive mycosis, but typical macroscopic findings with corresponding cultural findings can also confirm the diagnosis. Systemic mycosis requires an early initiation of a consistent antimycotic therapy as well as definitive surgical eradication of the focus in order to reduce high lethal rate. Resistances or incorrect dosages can be validated objectively by means of histological monitoring of the antimycotic therapy, thus affording early recognition of the need to change the substance class. PMID- 15826284 TI - [Invasive fungal infections in patients after liver transplantation]. AB - Advances in surgical technique, immunosuppression, and medical management have greatly improved clinical results after liver transplantation (LTx). Fungal infections in LTx-patients still represent serious complications and are associated with a significant decrease in survival. The majority of fungal infections in LTx-patients are caused by Candida species, which is explained by the major abdominal surgery. Aspergillus infections are second common, whereas other fungal infections such as pneumocystosis, cryptococcosis, or zygomycosis represent rare events. The high mortality of invasive fungal infections in LTx recipients is explained by the severity of the underlying medical condition and by difficulties in diagnosis and medical therapy. Currently available diagnostic tests do not allow a timely and reliable diagnosis of invasive fungal infections in LTx-patients. Amphotericin B has been the standard treatment for invasive candidiasis and aspergillosis for many years but the high frequency of side effects limits its application. Fluconazole is widely used due to better tolerability and fewer drug interactions. Disadvantages are the lack of activity against Aspergillus species and the selection of resistant Candida strains. Progress is to be expected from new antimycotic agents belonging to azoles (voriconazole) and echinocandins (caspofungin) as these are less toxic and have a broad range of antimycotic activity. Analysis of prognostic factors allows identifying LTx-patients at high risk for invasive fungal infection. Antimycotic prophylaxis or pre-emptive therapy may improve clinical outcome in this patient subgroup. PMID- 15826285 TI - [Fungal infections in patients with necrotizing pancreatitis: risk-factors, incidence, therapy]. AB - Haemorraghic necrotizing pancreatitis may lead to a serious disease with multi organ failure, which is to be treated with intensive care. Patients suffering from infected necrosis are usually operated (necrosectomy). By doing this, it is possible to get a microbiological analysis. The most common virulent species are Enterobacteriaceae. According to the literature, fungal infections appear in 15 30% of the cases. Since 1996, 73 patients were treated surgically in our department. A number of 50 patients (68,5%) developed a fungal infection during the course of the disease. The mortality rate was 62%. PMID- 15826286 TI - [Complicated course of oesophageal perforations because of fungal infections]. AB - Perforations of the oesophagus are characterized by a high mortality rate, varying between 7 and 49%. In the analyzed group of patients between the years 1986 and 2003, the mortality rate of 28% was caused by septic complications such as mediastinitis and pneumonia. Microbiological examinations of samples from different locations and various secretions, detected mycoses in 28% of the patients (n = 16). Compared with the total of mycoses, a higher mortality of 50% was calculated. The invasive Candida infection of the oesophagus itself can cause a perforation. In perforations of the oesophagus, simultaneous mycoses arise by fungi invading sterile compartments or by haematogenous and lymphogenous dissemination up to sepsis. Uncomplicated courses do not need antimycotic therapy. At the beginning of the treatment, a microbiological monitoring should be provided, particularly with regard to the intraoperative facts. In relation to the course and the risk factors of the patients, an antimycotic therapy is initiated. Surgical clearing and sufficient drainage of the collateral compartments such as pleural and mediastinal compartments is highly significant. PMID- 15826287 TI - [Surgical treatment of pulmonary aspergilloma]. AB - Invasive pulmonary aspergillosis is a life-threatening disease, developing from infection mostly by Aspergillus fumigatus. It is an opportunistic infection and occurs mostly in immunosuppressed patients, after tuberculosis, in patients with AIDS or malignomas. The most common symptom and complication is hemoptysis up to massive bleeding. Surgical treatment is obligatory on patients with a pulmonary aspergilloma. Time and form of intervention should be discussed between surgeon and pulmonologist. Resection should follow anatomical borders like lobectomy or pneumectomy. Any remaining intrathoracic cavities should be filled either with remaining lung tissue or with muscle flaps of latissimus or pectoralis muscle or omentum majus. Following these rules, intervention can be performed with reasonable mortality and morbidity rate. PMID- 15826288 TI - [Surveillance of invasive mold infections in lung transplant recipients: effect of antimycotic prophylaxis with itraconazole and voriconazole]. AB - Between January 2002 and December 2003 all 157 patients (pts) that underwent lung transplantation (LTx) at our institution were prospectively screened for invasive aspergillosis (IA) during their perioperative hospital stay. Patients were regarded as IA positive, if they met the EORTC criteria for 'probable' or 'proven' IA. Records of pts were screened retrospectively for antimycotic prophylaxis. Eight of the 157 pts developed 'probable' or 'proven' IA (5.1%) within 17 +/- 10 days after LTx. This was associated with a 14-fold increased mortality compared with all pts without aspergillosis (P < 0.01, OR 13.8, CI(95%) 2.5-82). Preoperative colonization with Aspergillus was a significant risk factor for IA (P < 0.001, OR 21.9, CI(95%) 4.9-97). We switched our prophylactic strategies to the primary administration of voriconazole in high risk pts (pre LTx colonization) starting in December 2002. Six pts (6%) of 101 pts receiving itraconazole for antimycotic prophylaxis beginning at postoperative day (POD) one developed IA, of which three pts showed cerebral aspergillosis. One pt (5%) of 18 pts receiving voriconazole prophylaxis developed IA, while 10 pts showed pretransplant colonization with Aspergillus species. Thirty-eight pts received itraconazole prophylaxis at a later time point (>POD 14). By switching our prophylactic strategy to the use of voriconazole in high risk pts, we have decreased the incidence of IA from 8% (six of 75) in 2002 to 2% (two of 82) in 2003. This study shows a high incidence of IA during the very early postoperative course after LTx of 5%. This is associated with a significantly increased risk for mortality. Voriconazole prophylaxis appears to be superior to itraconazole, especially in high risk pts with pretransplant Aspergillus colonization. PMID- 15826289 TI - [Antimycotic drugs under continuous renal replacement therapy]. AB - In critically ill patients with acute renal failure and continuous renal replacement therapy (CRRT), the elimination of antimycotic agents is influenced by many factors; these include not only the nature and dose of the renal replacement therapy but also the properties of the medicine. High protein binding, a high molecular weight and little or no renal elimination even in subjects with normal renal function suggest that a substance is not eliminated by CRRT. This is the case with amphotericin B, caspofungin, itraconazole and voriconazole, but not with fluconazole. In the case of fluconazole, an increase in dose than that used in patients with normal renal function may be required according to the nature and dose of CRRT, because the clearance of fluconazole can be substantially higher under CRRT than in subjects with normal renal function. Daily doses of 800 mg or more may be necessary. As a result of the diverse influences on elimination in patients receiving CRRT, the determination of fluconazole through concentrations cannot be dispensed with, in order to avoid underdosing and a failure of the antimycotic therapy. PMID- 15826290 TI - [Therapy of intraabdominal fungal infections]. AB - Candida albicans is the fourth most germ that can be identified on surgical intensive care unit (SICU). During the course of severe peritonitis recognition of Candida is crucial for physicians but interpretation of Candida-positive microbiologic samples is difficult. The indication for antimycotic therapy requires differentiation between harmless contamination or severe invasive mycosis associated with high mortality. Therefore, we propose a four-stage classification. Stage I is the initial contamination of the abdominal cavity by Candida spp. Stage IIa is characterized by persistence of fungi in patients without risk factors, IIb with risk factors respectively. Stage III means histological evidence of Candida invasion into the peritoneal layer. Stage IV is a generalized infection with fungemia/fungal sepsis. We recommend antimycotic therapy in stage IIb or higher. PMID- 15826291 TI - [Missing granulocytic infiltrate in pityriasis versicolor--indication of specific anti-inflammatory activity of the pathogen?]. AB - The yeast Malassezia furfur is a part of the resident flora of human skin. It causes various diseases such as pityriasis versicolor, which hardly shows signs of inflammation despite marked clinical symptoms (e.g. hypopigmentation). The pathophysiology related morphological picture might give a clue to this phenomenon. As a part of the literature data are controversial, the present study compared the inflammatory infiltrate of pityriasis versicolor with that of tinea corporis in 40 human skin preparations each from diagnostic specimens. All preparations were stained with HE and PAS. Neutrophilic granulocytes were counted in the HE stain, and hyphae and spores in the PAS stain. The number of counted cells was related to the size of the respective area and the values were compared between pityriasis and tinea corporis. Significantly, more neutrophilic granulocytes were found with tinea corporis (P > 0.01), while they were virtually not demonstrable with pityriasis versicolor. It is surprising that fungal load in the stratum corneum is significantly higher with pityriasis versicolor (P > 0.01). Obviously the immune response involving neutrophilic granulocytes does not occur despite high bacterial load. This might be explained by reduced immunogenicity because of high content of lipids in the cell membrane. Furthermore, pityriarubins that are produced during tryptophan metabolism might be involved, which, in a stimulus-dependent manner, can suppress the ROS production of neutrophilic granulocytes in vivo. PMID- 15826292 TI - [Fungal sepsis--therapeutical strategy]. AB - The incidence of fungal infection as well as fungal sepsis has increased dramatically during the last decade. Changes of local microbial flora after broad spectrum antibiotic therapy allow overgrowth of Candida species. Prophylactic strategies to lower fungal infection and sepsis include adequate and restrictive antibiotic therapy. Concerning the treatment of the septic syndrome, supportive as well as adjunctive strategies like early-goal-directed cardiovascular therapy, hydrocortisone replacement therapy, intense insulin application to achieve normoglycemia as well as the application of activated Protein C besides a consequent source control regimen and standard intensive care therapy, are able to improve significantly the outcome of septic patients. PMID- 15826293 TI - [Severe case of Candida peritonitis in a patient on CAPD--a successful treatment]. AB - Continuous ambulatory peritoneal dialysis (CAPD) is a widespread method of treatment used in approximately 10% of all patients suffering from terminal renal insufficiency. The main problem of this procedure is the increased risk of peritoneal infection. The incidence of such a peritonitis is quoted at one episode per 13-18.4 months of treatment. Candida peritonitis is a particularly severe form of CAPD peritonitis. This is a nosocomial infection with a high lethality rate of about 60%. The incidence of Candida peritonitis in CAPD patients amounts to approximately 5% of all intraabdominal infections and is on the increase. The authors describe a severe case of Candida peritonitis in a patient on CAPD. The therapeutic concept of this severe illness is illustrated. PMID- 15826294 TI - [Therapy on mycoses after simultaneous pancreas-kidney transplantation]. AB - Patients suffering from type I diabetes mellitus have a descending expectancy of life, after developing diabetic nephropathy and undergoing haemodialysis. After 3 years of haemodialysis, approximately only 40% of the patients are still alive. The only remedy of this incurable metabolic disease is the simultaneous pancreas kidney transplantation. After transplantation, fungal colonization and fungal infection is a serious threat for the patient's life. In this context, it is necessary to distinguish between colonization and invasive mycosis. In the case of unsuccessful antimycotic treatment, the removal of the transplanted organ, depending on the patient's condition, may be necessary to save the patient's life. PMID- 15826295 TI - [Indication for antimycotic therapy for tracheobronchial candidosis under artificial ventilation]. AB - Tracheobronchial candidosis is an impetuous complication in intensive care medicine. This article presents a concept to compare diagnostic procedure, Candida species and resistant species of different intensive care units with each other. This concept should encourage bench marking between similar intensive care units. The report and retrospective analysis of the intensive care course offer the opportunity to reflect own decisions and to adjust them to the current therapy strategies. Both procedures should improve the antimycotic therapy for intensive care units and should avoid the occurrence of resistant species. Candida species are often detected in the respiratory system of ventilated patients in intensive care, but this alone is no indication for antimycotic therapy. A strict retention is recommended, but this retention is diminished by an unclear infection, critical situation of the patient in the case of multiple organ failure, additional infection and long term ventilation. A therapy strategy for individual situations should be established and a close diagnostic procedure should be performed. A positive blood culture or detection of Candida species in two or more diagnostic materials indicate an early antimycotic therapy. PMID- 15826296 TI - [Recurrent Candida sepsis with prolonged respiratory failure and severe liver dysfunction]. AB - Systemic mycoses, especially pulmonary diseases and septicemia are observed increasingly at intensive care units. Essential risk factors for development of candidosis are the expanded use of antibiotics and immunocompromised patients, caused either as a result of a severe underlying disease or iatrogenically induced after organ transplantation. Candida albicans is the most frequent pathogen in microbiological findings. Blood cultures are only positive in massive fungemia. We report a 50-year-old patient with recurrent Candida-septicemia: rupture of the distal esophagus after dilatation because of cardiac achalasia with mediastinal emphysema and mediastinitis. Severe acute respiratory distress syndrome after aspiration with septic shock and acute renal failure at the beginning. Long-term mechanical ventilation, continuous renal replacement therapy and multifarious antibiotic therapy. Early microbiological samples of several positive blood cultures and bronchoalveolar lavages revealed the presence of Candida albicans. In the further clinical course, detection of Pseudomonas species in bronchoalveolar lavages and Staphylococci as well as Enterococci in a number of positive blood cultures. Later on development of a severe liver dysfunction with test results that showed an intrahepatic cholestasis. Because of coagulation failure commencement of artificial liver support with the MARS-system (molecule adsorbent recirculating system). Decrease of high bilirubin levels was accompanied by improvement of clinical condition of the patient. In the following course, repeated severe systemic infections with phases of septicemia or rather septic shock and detection of Candida in several positive blood cultures and bronchoalveolar lavages. In each case increasing bilirubin levels with signs of intrahepatic cholestasis and each time improvement with antimycotic therapy (voriconazol, caspofungin and fluconazol). The patient showed more and more signs of immunodeficiency in the sequel. The clinical appearance of candidosis is manifold. Systemic Candida infections are frequent in patients with immunodeficiency. A recurrent Candida septicemia with prolonged respiratory failure and severe liver dysfunction in form of cholestatic hepatosis, that improved several times with antimycotic therapy in combination with evidence based intensive care measures and artificial organ support is a comparatively rare event. PMID- 15826297 TI - [Pulmonary zygomycosis--a rare angioinvasive fungal infection]. AB - Zygomycosis caused by Rhizopus species is an aggressive and rapidly progressive opportunistic fungal infection in immunocompromised patients. It comprises mucocutaneous, rhinocerebral, pulmonary, urological and disseminated infections. Predisposing factors are immunosuppression owing to severe diseases, immunological defects or metabolic disturbances like diabetic ketoacidosis. Rhizopus infections are characterized by angioinvasive growth, necroses of infected tissue and perineural invasion. The histopathologic demonstrable invasion of blood vessels is remarkable for a fungal infection. The mortality of zygomycosis is very high, especially for disseminated disease and when immunosuppression cannot be corrected. We report about two cases of pulmonary zygomycosis, caused by Rhizopus spp.: patient 1, female 73 years old: Delayed clinical course according to hip arthroplasty infection and infection of a femoropopliteal bypass of the right leg, eventually exarticulation of the right hip joint, Pseudomonas pneumonia, severe sepsis caused by staphylococci, acute respiratory distress syndrome (ARDs), acute renal failure and multiple use of antibiotics. Subsequently detection of Rhizopus spp. in the bronchoalveolar lavage and treatment with amphotericin B for this reason. Patient 2, male 68 years old: transplantation of kidney in past medical history, presenting with acute renal failure and with quite a few infections before. In the sequel development of abscessing pneumonia on the right side with a pleural empyema. Rhizopus spp. were detected by microbiological testing in the empyema fluid. These findings required surgical intervention, resection of the lower lobe of the right lung and within the same operation of the renal graft because of rejection. The patient was treated with caspofungin. The further course was delayed by several septic phases. Both patients died later on in spite of all efforts. The very rarely seen pulmonary zygomycosis caused by infection with Rhizopus spp. developed in both patients owing to immunosuppression, in one patient iatrogenically induced by immunosuppressive drugs after organ transplantation, in the other as a result of prolonged severe sepsis. In comparison with other mycoses treatment of Rhizopus infections remains difficult. The affinity to blood vessels, where the fungi multiply, and their feature of vascular invasion with thrombosis and infarction complicate therapeutic efforts. PMID- 15826298 TI - 'PACLIMS': a component LIM system for high-throughput functional genomic analysis. AB - BACKGROUND: Recent advances in sequencing techniques leading to cost reduction have resulted in the generation of a growing number of sequenced eukaryotic genomes. Computational tools greatly assist in defining open reading frames and assigning tentative annotations. However, gene functions cannot be asserted without biological support through, among other things, mutational analysis. In taking a genome-wide approach to functionally annotate an entire organism, in this application the approximately 11,000 predicted genes in the rice blast fungus (Magnaporthe grisea), an effective platform for tracking and storing both the biological materials created and the data produced across several participating institutions was required. RESULTS: The platform designed, named PACLIMS, was built to support our high throughput pipeline for generating 50,000 random insertion mutants of Magnaporthe grisea. To be a useful tool for materials and data tracking and storage, PACLIMS was designed to be simple to use, modifiable to accommodate refinement of research protocols, and cost-efficient. Data entry into PACLIMS was simplified through the use of barcodes and scanners, thus reducing the potential human error, time constraints, and labor. This platform was designed in concert with our experimental protocol so that it leads the researchers through each step of the process from mutant generation through phenotypic assays, thus ensuring that every mutant produced is handled in an identical manner and all necessary data is captured. CONCLUSION: Many sequenced eukaryotes have reached the point where computational analyses are no longer sufficient and require biological support for their predicted genes. Consequently, there is an increasing need for platforms that support high throughput genome-wide mutational analyses. While PACLIMS was designed specifically for this project, the source and ideas present in its implementation can be used as a model for other high throughput mutational endeavors. PMID- 15826299 TI - Hepatocyte growth factor (HGF) in fecal samples: rapid detection by surface plasmon resonance. AB - BACKGROUND: The development of biosensors, based on surface plasmon resonance (SPR) technology, enables monitoring of a variety of biospecific interactions without the need for chemical-, biological- or radiological-labelled reagents. METHOD: We utilised SPR to detect hepatocyte growth factor (HGF) in reconstituted faecal samples and studied samples from patients with infectious gastroenteritis (n = 20) and normal controls (n = 10). Mouse anti-human HGF monoclonal antibodies and recombinant human HGF receptor (c-Met)/Fc chimera were immobilised in flow cells of a CM5 biosensor chip. RESULTS: We found that infectious gastroenteritis produced a higher signal response compared to controls, due to binding of HGF to monoclonal anti-HGF antibody as well as binding of HGF to c-Met receptor (p < 0.01). The SPR signal response correlated with results from ELISA (r = 72%, p > 0.001). The signal response decreased significantly (p < 0.05) when samples were diluted with dextran, because of reduction in both specific as well as unspecific binding of HGF to dextran. The decrease in the specific response might imply that the dextran- binding site for HGF overlaps with the antibody binding epitope, or that dextran binding induces a conformational change of the HGF molecule. Bands corresponding to HGF were found by gel electrophoresis of purified faeces in an affinity chromatography column immobilised by HGF ligands. CONCLUSION: Determination of HGF by SPR might be beneficial in diagnosis of acute situations that present with symptoms of gastroenteritis and may, possibly, guide appropriate medical treatments. This is to our knowledge the first report on the use of SPR for detection of HGF in faeces samples. PMID- 15826300 TI - The role of Mediterranean diet in the epidemiology of metabolic syndrome; converting epidemiology to clinical practice. AB - Metabolic syndrome is a collection of associated conditions such as dyslipidemia, high blood pressure, impaired glucose tolerance and tendency to develop fat around the abdomen. It is now well known that individuals with the metabolic syndrome are at high risk for atherosclerosis and, especially, coronary heart disease. However, it has been suggested that people with the metabolic syndrome may benefit from aggressive lifestyle modification, through diet and exercise. In this review we summarize scientific evidence regarding the effect of Mediterranean diet on the development of metabolic syndrome. PMID- 15826301 TI - Halothane potentiates the alcohol-adduct induced TNF-alpha release in heart endothelial cells. AB - BACKGROUND: The possibility exists for major complications to occur when individuals are intoxicated with alcohol prior to anesthetization. Halothane is an anesthetic that can be metabolized by the liver into a highly reactive product, trifluoroacetyl chloride, which reacts with endogenous proteins to form a trifluoroacetyl-adduct (TFA-adduct). The MAA-adduct which is formed by acetaldehyde (AA) and malondialdehyde reacting with endogenous proteins, has been found in both patients and animals chronically consuming alcohol. These TFA and MAA-adducts have been shown to cause the release of inflammatory products by various cell types. If both adducts share a similar mechanism of cell activation, receiving halothane anesthesia while intoxicated with alcohol could exacerbate the inflammatory response and lead to cardiovascular injury. METHODS: We have recently demonstrated that the MAA-adduct induces tumor necrosis factor-alpha (TNF-alpha) release by heart endothelial cells (HECs). In this study, pair and alcohol-fed rats were randomized to receive halothane pretreatments intra peritoneal. Following the pretreatments, the intact heart was removed, HECs were isolated and stimulated with unmodified bovine serum albumin (Alb), MAA-modified Alb (MAA-Alb), Hexyl-MAA, or lipopolysaccharide (LPS), and supernatant concentrations of TNF-alpha were measured by ELISA. RESULTS: Halothane pre treated rat HECs released significantly greater TNF-alpha concentration following MAA-adduct and LPS stimulation than the non-halothane pre-treated in both pair and alcohol-fed rats, but was significantly greater in the alcohol-fed rats. CONCLUSION: These results demonstrate that halothane and MAA-adduct pre-treatment increases the inflammatory response (TNF-alpha release). Also, these results suggest that halothane exposure may increase the risk of alcohol-induced heart injury, since halothane pre-treatment potentiates the HEC TNF-alpha release measured following both MAA-Alb and LPS stimulation. PMID- 15826302 TI - Monitoring processed, mature Human Immunodeficiency Virus type 1 particles immediately following treatment with a protease inhibitor-containing treatment regimen. AB - Protease inhibitors (PIs) block HIV-1 maturation into an infectious virus particle by inhibiting the protease processing of gag and gag-pol precursor proteins. We have used a simple anti-HIV-1 p24 Western blot to monitor the processing of p55gag precursor into the mature p24 capsid immediately following the first dosage of a PI-containing treatment regimen. Evidence of PI activity was observed in plasma virus as early as 72 hours post treatment-initiation and was predictive of plasma viral RNA decrease at 4 weeks. PMID- 15826303 TI - Breakdown of accommodation in nerve: a possible role for persistent sodium current. AB - BACKGROUND: Accommodation and breakdown of accommodation are important elements of information processing in nerve fibers, as they determine how nerve fibers react to natural slowly changing stimuli or electrical stimulation. The aim of the present study was to elucidate the biophysical mechanism of breakdown of accommodation, which at present is unknown. RESULTS: A model of a space-clamped motor nerve fiber was developed. It was found that this new model could reproduce breakdown of accommodation when it included a low-threshold, rapidly activating, persistent sodium current. However, the phenomenon was not reproduced when the persistent sodium current did not have fast activation kinetics or a low activation threshold. CONCLUSION: The present modeling study suggests that persistent, low-threshold, rapidly activating sodium currents have a key role in breakdown of accommodation, and that breakdown of accommodation can be used as a tool for studying persistent sodium current under normal and pathological conditions. PMID- 15826304 TI - Myeloid antigens in childhood lymphoblastic leukemia: clinical data point to regulation of CD66c distinct from other myeloid antigens. AB - BACKGROUND: Aberrant expression of myeloid antigens (MyAgs) on acute lymphoblastic leukemia (ALL) cells is a well-documented phenomenon, although its regulating mechanisms are unclear. MyAgs in ALL are interpreted e.g. as hallmarks of early differentiation stage and/or lineage indecisiveness. Granulocytic marker CD66c -- Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) is aberrantly expressed on ALL with strong correlation to genotype (negative in TEL/AML1 and MLL/AF4, positive in BCR/ABL and hyperdiploid cases). METHODS: In a cohort of 365 consecutively diagnosed Czech B-precursor ALL patients, we analyze distribution of MyAg+ cases and mutual relationship among CD13, CD15, CD33, CD65 and CD66c. The most frequent MyAg (CD66c) is studied further regarding its stability from diagnosis to relapse, prognostic significance and regulation of surface expression. For the latter, flow cytometry, Western blot and quantitative RT-PCR on sorted cells is used. RESULTS: We show CD66c is expressed in 43% patients, which is more frequent than other MyAgs studied. In addition, CD66c expression negatively correlates with CD13 (p < 0.0001), CD33 (p = 0.002) and/or CD65 (p = 0.029). Our data show that different myeloid antigens often differ in biological importance, which may be obscured by combining them into "MyAg positive ALL". We show that unlike other MyAgs, CD66c expression is not shifted from the onset of ALL to relapse (n = 39, time to relapse 0.3-5.3 years). Although opposite has previously been suggested, we show that CEACAM6 transcription is invariably followed by surface expression (by quantitative RT PCR on sorted cells) and that malignant cells containing CD66c in cytoplasm without surface expression are not found by flow cytometry nor by Western blot in vivo. We report no prognostic significance of CD66c, globally or separately in genotype subsets of B-precursor ALL, nor an association with known risk factors (n = 254). CONCLUSION: In contrast to general notion we show that different MyAgs in lymphoblastic leukemia represent different biological circumstances. We chose the most frequent and tightly genotype-associated MyAg CD66c to show its stabile expression in patients from diagnosis to relapse, which differs from what is known on the other MyAgs. Surface expression of CD66c is regulated at the gene transcription level, in contrast to previous reports. PMID- 15826305 TI - Statistical Viewer: a tool to upload and integrate linkage and association data as plots displayed within the Ensembl genome browser. AB - BACKGROUND: To facilitate efficient selection and the prioritization of candidate complex disease susceptibility genes for association analysis, increasingly comprehensive annotation tools are essential to integrate, visualize and analyze vast quantities of disparate data generated by genomic screens, public human genome sequence annotation and ancillary biological databases. We have developed a plug-in package for Ensembl called "Statistical Viewer" that facilitates the analysis of genomic features and annotation in the regions of interest defined by linkage analysis. RESULTS: Statistical Viewer is an add-on package to the open source Ensembl Genome Browser and Annotation System that displays disease study specific linkage and/or association data as 2 dimensional plots in new panels in the context of Ensembl's Contig View and Cyto View pages. An enhanced upload server facilitates the upload of statistical data, as well as additional feature annotation to be displayed in DAS tracts, in the form of Excel Files. The Statistical View panel, drawn directly under the ideogram, illustrates lod score values for markers from a study of interest that are plotted against their position in base pairs. A module called "Get Map" easily converts the genetic locations of markers to genomic coordinates. The graph is placed under the corresponding ideogram features a synchronized vertical sliding selection box that is seamlessly integrated into Ensembl's Contig- and Cyto- View pages to choose the region to be displayed in Ensembl's "Overview" and "Detailed View" panels. To resolve Association and Fine mapping data plots, a "Detailed Statistic View" plot corresponding to the "Detailed View" may be displayed underneath. CONCLUSION: Features mapping to regions of linkage are accentuated when Statistic View is used in conjunction with the Distributed Annotation System (DAS) to display supplemental laboratory information such as differentially expressed disease genes in private data tracks. Statistic View is a novel and powerful visual feature that enhances Ensembl's utility as valuable resource for integrative genomic-based approaches to the identification of candidate disease susceptibility genes. At present there are no other tools that provide for the visualization of 2-dimensional plots of quantitative data scores against genomic coordinates in the context of a primary public genome annotation browser. PMID- 15826306 TI - Proliferation dynamics of germinative zone cells in the intact and excitotoxically lesioned postnatal rat brain. AB - BACKGROUND: The forebrain subventricular zone (SVZ)-olfactory bulb pathway and hippocampal subgranular zone (SGZ) generate neurons into adulthood in the mammalian brain. Neurogenesis increases after injury to the adult brain, but few studies examine the effect of injury on neural and glial precursors in the postnatal brain. To characterize the spatio-temporal dynamics of cell proliferation in the germinative zones, this study utilized a model of postnatal damage induced by NMDA injection in the right sensorimotor cortex at postnatal day 9. Dividing cell populations were labeled with 5-Bromodeoxyuridine (BrdU) in the intact and damaged postnatal brain. Identity of proliferating cells was determined by double immunolabeling with nestin, GFAP, NeuN and tomato lectin (TL). RESULTS: In the control brain, grouped BrdU+ cells were observed in the Rostral Migratory Stream (RMS), SVZ and SGZ. Maximal proliferation was seen at P12, persisted until P23 and diminished by P49. After injury, a striking reduction in the number of BrdU+ cells was observed in the ipsilateral SVZ from 10 hours (58% decrease) until 14 days post-lesion (88% decrease). In contrast, an increase in grouped BrdU+ cells was seen in the striatum adjacent to the depleted SVZ. Significantly reduced numbers of BrdU+ cells were also seen in the RMS until 3 days post-lesion. No changes were noted in the SGZ. Both in controls and lesioned hemispheres, BrdU+ cells located in the germinal zones were mostly nestin positive and negative for GFAP, NeuN, and TL. In the SVZ area lining the ventricle, BrdU+/nestin+ cells were mainly located between TL+ ependyma and parenchymal GFAP+ astrocytes. After excitotoxicity, a decrease in the number and orientation of GFAP/nestin+ prolongations leaving the SVZ to the cortex, corpus callosum and striatum was noted until 5 days post-lesion. CONCLUSION: Postnatal excitotoxic injury differentially affects proliferating cells in the germinative zones: no change is observed in the dentate gyrus whereas excitotoxicity causes a significant decrease in proliferating cells in the SVZ and RMS. Depletion of BrdU+ cells in the postnatal SVZ and RMS differs from previous studies after adult brain injury and may affect the SVZ-RMS migration and is suggestive of progenitor recruitment to injured areas. PMID- 15826307 TI - A strategy to study tyrosinase transgenes in mouse melanocytes. AB - BACKGROUND: A number of transgenic mice carrying different deletions in the Locus Control Region (LCR) of the mouse tyrosinase (Tyr) gene have been developed and analysed in our laboratory. We require melanocytes from these mice, to further study, at the cellular level, the effect of these deletions on the expression of the Tyr transgene, without potential interference with or from the endogenous Tyr alleles. It has been previously reported that it is possible to obtain and immortalize melanocyte cell cultures from postnatal mouse skin. RESULTS: Here, we describe the efforts towards obtaining melanocyte cultures from our Tyr transgenic mice. We have bred our Tyr transgenic mice into Tyr c-32DSD mutant background, lacking the endogenous Tyr locus. In these conditions, we failed to obtain immortalized melanocytes. We decided to include the inactivation of the Ink4a-Arf locus to promote melanocyte immortalisation. For this purpose, we report the segregation of the Ink4a-Arf null allele from the brown (Tyrp1b) mutation in mice. Finally, we found that Ink4a-Arf +/- and Ink4a-Arf -/- melanocytes had undistinguishable tyrosine hydroxylase activities, although the latter showed reduced cellular pigmentation content. CONCLUSION: The simultaneous presence of precise genomic deletions that include the tyrosinase locus, such as the Tyr c-32DSD allele, the Tyr transgene itself and the inactivated Ink4a-Arf locus in Tyrp1B genetic background appear as the crucial combination to perform forthcoming experiments. We cannot exclude that Ink4a-Arf mutations could affect the melanin biosynthetic pathway. Therefore, subsequent experiments with melanocytes will have to be performed in a normalized genetic background regarding the Ink4a-Arf locus. PMID- 15826308 TI - Subjective face recognition difficulties, aberrant sensibility, sleeping disturbances and aberrant eating habits in families with Asperger syndrome. AB - BACKGROUND: The present study was undertaken in order to determine whether a set of clinical features, which are not included in the DSM-IV or ICD-10 for Asperger Syndrome (AS), are associated with AS in particular or whether they are merely a familial trait that is not related to the diagnosis. METHODS: Ten large families, a total of 138 persons, of whom 58 individuals fulfilled the diagnostic criteria for AS and another 56 did not to fulfill these criteria, were studied using a structured interview focusing on the possible presence of face recognition difficulties, aberrant sensibility and eating habits and sleeping disturbances. RESULTS: The prevalence for face recognition difficulties was 46.6% in individuals with AS compared with 10.7% in the control group. The corresponding figures for subjectively reported presence of aberrant sensibilities were 91.4% and 46.6%, for sleeping disturbances 48.3% and 23.2% and for aberrant eating habits 60.3% and 14.3%, respectively. CONCLUSION: An aberrant processing of sensory information appears to be a common feature in AS. The impact of these and other clinical features that are not incorporated in the ICD-10 and DSM-IV on our understanding of AS may hitherto have been underestimated. These associated clinical traits may well be reflected by the behavioural characteristics of these individuals. PMID- 15826309 TI - PREP-Mt: predictive RNA editor for plant mitochondrial genes. AB - BACKGROUND: In plants, RNA editing is a process that converts specific cytidines to uridines and uridines to cytidines in transcripts from virtually all mitochondrial protein-coding genes. There are thousands of plant mitochondrial genes in the sequence databases, but sites of RNA editing have not been determined for most. Accurate methods of RNA editing site prediction will be important in filling in this information gap and could reduce or even eliminate the need for experimental determination of editing sites for many sequences. Because RNA editing tends to increase protein conservation across species by "correcting" codons that specify unconserved amino acids, this principle can be used to predict editing sites by identifying positions where an RNA editing event would increase the conservation of a protein to homologues from other plants. PREP-Mt takes this approach to predict editing sites for any protein-coding gene in plant mitochondria. RESULTS: To test the general applicability of the PREP-Mt methodology, RNA editing sites were predicted for 370 full-length or nearly full length DNA sequences and then compared to the known sites of RNA editing for these sequences. Of 60,263 cytidines in this test set, PREP-Mt correctly classified 58,994 as either an edited or unedited site (accuracy = 97.9%). PREP Mt properly identified 3,038 of the 3,698 known sites of RNA editing (sensitivity = 82.2%) and 55,956 of the 56,565 known unedited sites (specificity = 98.9%). Accuracy and sensitivity increased to 98.7% and 94.7%, respectively, after excluding the 489 silent editing sites (which have no effect on protein sequence or function) from the test set. CONCLUSION: These results indicate that PREP-Mt is effective at identifying C to U RNA editing sites in plant mitochondrial protein-coding genes. Thus, PREP-Mt should be useful in predicting protein sequences for use in molecular, biochemical, and phylogenetic analyses. In addition, PREP-Mt could be used to determine functionality of a mitochondrial gene or to identify particular sequences with unusual editing properties. The PREP-Mt methodology should be applicable to any system where RNA editing increases protein conservation across species. PMID- 15826311 TI - Who wants to join preventive trials?--Experience from the Estonian Postmenopausal Hormone Therapy Trial [ISRCTN35338757]. AB - BACKGROUND: The interest of patients in participating in randomized clinical trials involving treatments has been widely studied, but there has been much less research on interest in preventive trials. The objective of this study was to find out how many women would be interested in a trial involving postmenopausal hormone therapy (PHT) and how the women's background characteristics and opinions correlated to their interest. METHODS: The data come from recruitment questionnaires (n = 2000) sent to women in Estonia in 1998. A random sample of women aged 45 to 64 was drawn from the Population Registry. The trial is a two group randomized trial comparing estrogen-progestogen therapy with placebo or no drugs. A brief description of the study was attached to the questionnaires. Women were not told at this stage of the recruitment which group they would be assigned to, however, they were told of the chance to receive either hormone, placebo or no treatment. RESULTS: After two reminders, 1312 women (66%) responded. Eleven percent of the women approached (17% of the respondents) were interested in joining the trial, and 8% wanted more information before deciding. When the 225 women who stated clearly that they were interested in joining and the 553 women who said they were not interested were compared, it was found that interested women were younger and, adjusting for age, that more had given birth; in other respects, the sociodemographic characteristics and health habits of the interested women were similar to those of the non-interested women. The interested women had made more use of more health services, calcium preparations and PHT, they were more often overweight, and more had chronic diseases and reported symptoms. Interested women's opinions on the menopause were more negative, and they favoured PHT more than the non-interested women. CONCLUSION: Unlike the situation described in previous reports on preventive trials, in this case Estonian women interested in participating in a PHT trial were not healthier than other women. This suggests that trials involving PHT are more similar to treatment trials than to preventive trials. In a randomized controlled trial, more information should be obtained from those women who decline to participate. PMID- 15826312 TI - Identification of a truncated nucleoprotein in avian metapneumovirus-infected cells encoded by a second AUG, in-frame to the full-length gene. AB - BACKGROUND: Avian metapneumoviruses (aMPV) cause an upper respiratory disease with low mortality, but high morbidity primarily in commercial turkeys. There are three types of aMPV (A, B, C) of which the C type is found only in the United States. Viruses related to aMPV include human, bovine, ovine, and caprine respiratory syncytial viruses and pneumonia virus of mice, as well as the recently identified human metapneumovirus (hMPV). The aMPV and hMPV have become the type viruses of a new genus within the Metapneumovirus. The aMPV nucleoprotein (N) amino acid sequences of serotypes A, B, and C were aligned for comparative analysis. Based on predicted antigenicity of consensus protein sequences, five aMPV-specific N peptides were synthesized for development of peptide-antigens and antisera. RESULTS: The presence of two aMPV nucleoprotein (N) gene encoded polypeptides was detected in aMPV/C/US/Co and aMPV/A/UK/3b infected Vero cells. Nucleoprotein 1 (N1) encoded from the first open reading frame (ORF) was predicted to be 394 amino acids in length for aMPV/C/US/Co and 391 amino acids in length for aMPV/A/UK/3b with approximate molecular weights of 43.3 kilodaltons and 42.7 kilodaltons, respectively. Nucleoprotein 2 (N2) was hypothesized to be encoded by a second downstream ORF in-frame with ORF1 and encoded a protein predicted to contain 328 amino acids for aMPV/C/US/Co or 259 amino acids for aMPV/A/UK/3b with approximate molecular weights of 36 kilodaltons and 28.3 kilodaltons, respectively. Peptide antibodies to the N-terminal and C terminal portions of the aMPV N protein confirmed presence of these products in both aMPV/C/US/Co- and aMPV/A/UK/3b-infected Vero cells. N1 and N2 for aMPV/C/US/Co ORFs were molecularly cloned and expressed in Vero cells utilizing eukaryotic expression vectors to confirm identity of the aMPV encoded proteins. CONCLUSION: This is the first reported identification of potential, accessory in frame N2 ORF gene products among members of the Paramyxoviridae. Genomic sequence analyses of related members of the Pneumovirinae other than aMPV, including human respiratory syncytial virus and bovine respiratory syncytial virus demonstrated the presence of this second potential ORF among these agents. PMID- 15826313 TI - Why nutraceuticals do not prevent or treat Alzheimer's disease. AB - A great deal of research has pointed to deleterious roles of metal ions in the development of Alzheimer's disease. These include: i) the precipitation and aggregation of amyloid beta (Abeta) peptides to form senile plaques and neurofibrillary tangles, and/or ii) the augmentation of oxidative stress by metal ion mediated production and activation of hydrogen peroxide. The growing trend in nutraceutical intake is in part a result of the belief that they postpone the development of dementias such as Alzheimer's disease. However, pathogenic events centred on metal ions are expected to be aggravated by frequent nutraceutical intake. Novel therapeutic approaches centred on chelators with specificity for copper and iron ions should be fully explored. PMID- 15826314 TI - Inhaled corticosteroids for abnormal pulmonary function in children with a history of chronic lung disease of infancy: study protocol [ISRCTN55153521]. AB - BACKGROUND: There is considerable evidence from the literature that children with chronic lung disease of infancy (CLD) have abnormal pulmonary function in childhood and this could have an impact on their life quality and overall health. There are similarities between CLD and asthma, and corticosteroids are the mainstay treatment for asthma. Many physicians use inhaled corticosteroids in children with CLD with no evidence. Therefore we wish to conduct a randomized double-blinded placebo controlled trial to test for the role of inhaled corticosteroids in children aged from 3 to 9 years with a history of CLD. Our primary hypothesis will be that inhaled corticosteroids are beneficial in children with CLD. METHODS: Our primary hypothesis is that using inhaled steroids; Beclomethasone Dipropionate (QVAR) 100 mcg 2 puffs 2 times a day for 6 weeks will improve the respiratory system resistance and the quality of life in children with CLD. DISCUSSION: We propose that Beclomethasone Dipropionate (QVAR) will affect the pulmonary function after 6 weeks of treatment. In summary we think that our study will highlight knowledge on whether the use of inhaled steroids is clinically effective for CLD. PMID- 15826315 TI - Assessing spatio-temporal variability of risk surfaces using residential history data in a case control study of breast cancer. AB - BACKGROUND: Most analyses of spatial clustering of disease have been based on either residence at the time of diagnosis or current residence. An underlying assumption in these analyses is that residence can be used as a proxy for environmental exposure. However, exposures earlier in life and not just those in the most recent period may be of significance. In breast cancer, there is accumulating evidence that early life exposures may contribute to risk. We explored spatio-temporal patterns of risk surfaces using data on lifetime residential history in a case control study of breast cancer, and identified elevated areas of risk and areas potentially having more exposure opportunities, defined as risk surfaces in this study. This approach may be more relevant in understanding the environmental etiology of breast cancer, since lifetime cumulative exposures or exposures at critical times may be more strongly associated with risk for breast cancer than exposures from the recent period. RESULTS: A GIS-based exploratory spatial analysis was applied, and spatio temporal variability of those risk surfaces was evaluated using the standardized difference in density surfaces between cases and controls. The significance of the resulting risk surfaces was tested and reported as p-values. These surfaces were compared for premenopausal and postmenopausal women, and were obtained for each decade, from the 1940s to 1990s. We found strong evidence of clustering of lifetime residence for premenopausal women (for cases relative to controls), and a less strong suggestion of such clustering for postmenopausal women, and identified a substantial degree of temporal variability of the risk surfaces. CONCLUSION: We were able to pinpoint geographic areas with higher risk through exploratory spatial analyses, and to assess temporal variability of the risk surfaces, thus providing a working hypothesis on breast cancer and environmental exposures. Geographic areas with higher case densities need further epidemiologic investigation for potential relationships between lifetime environmental exposures and breast cancer risk. Examination of lifetime residential history provided additional information on geographic areas associated with higher risk; limiting exploration of chronic disease clustering to current residence may neglect important relationships between location and disease. PMID- 15826316 TI - Phase III trial of postoperative cisplatin, interferon alpha-2b, and 5-FU combined with external radiation treatment versus 5-FU alone for patients with resected pancreatic adenocarcinoma -- CapRI: study protocol [ISRCTN62866759]. AB - After surgical intervention with curative intention in specialised centres the five-year survival of patients with carcinoma of the exocrine pancreas is only 15%. The ESPAC-1 trial showed an increased five-year survival of 21% achieved with adjuvant chemotherapy. Investigators from the Virginia Mason Clinic have reported a 5-year survival rate of 55% in a phase II trial evaluating adjuvant chemotherapy, immunotherapy and external-beam radiation. DESIGN: The CapRI study is an open, controlled, prospective, randomised multi-centre phase III trial. Patients in study arm A will be treated as outpatients with 5-Fluorouracil; Cisplatin and 3 million units Interferon alpha-2b for 5 1/2 weeks combined with external beam radiation. After chemo-radiation the patients receive continuous 5 FU infusions for two more cycles. Patients in study arm B will be treated as outpatients with intravenous bolus injections of folinic acid, followed by intravenous bolus injections of 5-FU given on 5 consecutive days every 28 days for 6 cycles. A total of 110 patients with specimen-proven R0 or R1 resected pancreatic adenocarcinoma will be enrolled. An interim analysis for patient safety reasons will be done one year after start of recruitment. Evaluation of the primary endpoint will be performed two years after the last patients' enrollment. DISCUSSION: The aim of this study is to evaluate the overall survival period attained by chemo-radiotherapy including interferon alpha 2b administration with adjuvant chemotherapy. The influence of interferon alpha on the effectiveness of the patients' chemoradiation regimen, the toxicity, the disease-free interval and the quality of life are analysed. Different factors are tested in terms of their potential role as predictive markers. PMID- 15826317 TI - Many accurate small-discriminatory feature subsets exist in microarray transcript data: biomarker discovery. AB - BACKGROUND: Molecular profiling generates abundance measurements for thousands of gene transcripts in biological samples such as normal and tumor tissues (data points). Given such two-class high-dimensional data, many methods have been proposed for classifying data points into one of the two classes. However, finding very small sets of features able to correctly classify the data is problematic as the fundamental mathematical proposition is hard. Existing methods can find "small" feature sets, but give no hint how close this is to the true minimum size. Without fundamental mathematical advances, finding true minimum size sets will remain elusive, and more importantly for the microarray community there will be no methods for finding them. RESULTS: We use the brute force approach of exhaustive search through all genes, gene pairs (and for some data sets gene triples). Each unique gene combination is analyzed with a few-parameter linear-hyperplane classification method looking for those combinations that form training error-free classifiers. All 10 published data sets studied are found to contain predictive small feature sets. Four contain thousands of gene pairs and 6 have single genes that perfectly discriminate. CONCLUSION: This technique discovered small sets of genes (3 or less) in published data that form accurate classifiers, yet were not reported in the prior publications. This could be a common characteristic of microarray data, thus making looking for them worth the computational cost. Such small gene sets could indicate biomarkers and portend simple medical diagnostic tests. We recommend checking for small gene sets routinely. We find 4 gene pairs and many gene triples in the large hepatocellular carcinoma (HCC, Liver cancer) data set of Chen et al. The key component of these is the "placental gene of unknown function", PLAC8. Our HMM modeling indicates PLAC8 might have a domain like part of lP59's crystal structure (a Non-Covalent Endonuclease lii-Dna Complex). The previously identified HCC biomarker gene, glypican 3 (GPC3), is part of an accurate gene triple involving MT1E and ARHE. We also find small gene sets that distinguish leukemia subtypes in the large pediatric acute lymphoblastic leukemia cancer set of Yeoh et al. PMID- 15826318 TI - The neuronal ceroid lipofuscinosis Cln8 gene expression is developmentally regulated in mouse brain and up-regulated in the hippocampal kindling model of epilepsy. AB - BACKGROUND: The neuronal ceroid lipofuscinoses (NCLs) are a group of inherited neurodegenerative disorders characterized by accumulation of autofluorescent material in many tissues, especially in neurons. Mutations in the CLN8 gene, encoding an endoplasmic reticulum (ER) transmembrane protein of unknown function, underlie NCL phenotypes in humans and mice. The human phenotype is characterized by epilepsy, progressive psychomotor deterioration and visual loss, while motor neuron degeneration (mnd) mice with a Cln8 mutation show progressive motor neuron dysfunction and retinal degeneration. RESULTS: We investigated spatial and temporal expression of Cln8 messenger ribonucleic acid (mRNA) using in situ hybridization, reverse transcriptase polymerase chain reaction (RT-PCR) and northern blotting. Cln8 is ubiquitously expressed at low levels in embryonic and adult tissues. In prenatal embryos Cln8 is most prominently expressed in the developing gastrointestinal tract, dorsal root ganglia (DRG) and brain. In postnatal brain the highest expression is in the cortex and hippocampus. Expression of Cln8 mRNA in the central nervous system (CNS) was also analyzed in the hippocampal electrical kindling model of epilepsy, in which Cln8 expression was rapidly up-regulated in hippocampal pyramidal and granular neurons. CONCLUSION: Expression of Cln8 in the developing and mature brain suggests roles for Cln8 in maturation, differentiation and supporting the survival of different neuronal populations. The relevance of Cln8 up-regulation in hippocampal neurons of kindled mice should be further explored. PMID- 15826319 TI - Lipoprotein docosapentaenoic acid is associated with serum matrix metalloproteinase-9 concentration. AB - BACKGROUND: Polyunsaturated fatty acids (PUFA) are thought to play important roles in inflammation. The n-3 series is considered as anti-inflammatory, and some studies have reported increased plasma n-3 polyunsaturated fatty acid pattern in chronic inflammatory conditions. In this study we sought to clarify relationships of the levels of arachidonic acid and the polyunsaturated n-3 fatty acid compositions of isolated LDL, HDL2 and HDL3 particles with matrix metalloproteinase-9 (MMP-9), a marker of inflammation. RESULTS: The subjects were divided into two groups: those with lower and those with higher than the median serum MMP-9 concentration. In all lipoprotein fractions, the mean percentage of docosapentaenoic acid (C22:5n-3) was higher in the group of subjects with higher MMP-9 level than in those with lower serum MMP-9 concentration (P < 0.01 for all). Likewise, the ratio of docosapentaenoic acid to arachidonic acid (C20:4n-6) was higher in the subjects with higher MMP-9 compared with the lower MMP-9 group (P < 0.001 for all). CONCLUSION: So far, the evidence for an anti-inflammatory role of the n-3 PUFA has come from dietary interventions. Our results were obtained from a free-living population and indicate that there is a positive correlation between n-3 docosapentaenoic acid and MMP-9. What had triggered the rise in MMP-9 is not known, since serum level of MMP-9 is raised in many inflammatory conditions. These findings may indicate an increased biosynthesis of n-3 polyunsaturated fatty acids in subclinical inflammation. PMID- 15826321 TI - Conceptions of objects across categories: childhood patterns resemble those of adults. AB - Studies of category-specific disorders have suggested that categories of living and non-living things vary in the properties that are most salient to recognition. Studies of the object features generated by normal adults have also revealed different patterns of responses to different categories. These adult patterns are likely to originate in childhood, but there are few reports of children's verbal conceptions of objects, and none at present of objects from different categories. This paper investigates the development of object conceptions, in a large group of children, aged 3 years 7 months to 11 years 6 months, in response to 'What is a -- ?' questions directed to 72 objects, selected from two categories of living things (animals and fruit/vegetables) and two categories of artifacts (implements and vehicles). Proportions of perceptual to-functional features provided by the children to living and non-living things varied with the range of features defined as functions, just as studies of adults have found. Apart from the distribution of superordinate responses, which were significantly more salient to living than to non-living categories, no other property separated the two categories. Only the category of implements could be distinguished from the other categories on the basis of the features generated. It is argued that the perceptual-functional theory of category-specific disorders receives little support from this study, but that in general the distribution of features generated by young children is similar to that produced by normal adults. PMID- 15826320 TI - Risk factors for delirium in acutely admitted elderly patients: a prospective cohort study. AB - BACKGROUND: Delirium is a neuropsychiatric syndrome frequently observed in elderly hospitalised patients and can be found in any medical condition. Due to the severe consequences, early recognition of delirium is important in order to start treatment in time. Despite the high incidence rate, the occurrence of delirium is not always identified as such. Knowledge of potential risk factors is important. The aim of the current study is to determine factors associated with the occurrence of a prevalent delirium among elderly patients acutely admitted to an internal medicine ward. METHODS: All consecutive patients of 65 years and over acutely admitted to the Department of Internal Medicine of the Academic Medical Centre, Amsterdam, a university hospital, were asked to participate. The presence of delirium was determined within 48 hrs after admission by an experienced geriatrician. RESULTS: In total, 126 patients were included, 29% had a prevalent delirium after acute admission. Compared to patients without delirium, patients with delirium were older, more often were cognitively and physically impaired, more often were admitted due to water and electrolyte disturbances, and were less often admitted due to malignancy or gastrointestinal bleeding. Independent risk factors for having a prevalent delirium after acute admission were premorbid cognitive impairment, functional impairment, an elevated urea nitrogen level, and the number of leucocytes. CONCLUSIONS: In this study, the most important independent risk factors for a prevalent delirium after acute admission were cognitive and physical impairment, and a high serum urea nitrogen concentration. These observations might contribute to an earlier identification and treatment of delirium in acutely admitted elderly patients. PMID- 15826322 TI - Why the spontaneous images created by the hands during talk can help make TV advertisements more effective. AB - The design of effective communications depends upon an adequate model of the communication process. The traditional model is that speech conveys semantic information and bodily movement conveys information about emotion and interpersonal attitudes. But McNeill (2000) argues that this model is fundamentally wrong and that some bodily movements, namely spontaneous hand movements generated during talk (iconic gestures), are integral to semantic communication. But can we increase the effectiveness of communication using this new theory? Focusing on advertising we found that advertisements in which the message was split between speech and iconic gesture (possible on TV) were significantly more effective than advertisements in which meaning resided purely in speech or language (radio/newspaper). We also found that the significant differences in communicative effectiveness were maintained across five consecutive trials. We compared the communicative power of professionally made TV advertisements in which a spoken message was accompanied either by iconic gestures or by pictorial images, and found the iconic gestures to be more effective. We hypothesized that iconic gestures are so effective because they illustrate and isolate just the core semantic properties of a product. This research suggests that TV advertisements can be made more effective by incorporating iconic gestures with exactly the right temporal and semantic properties. PMID- 15826323 TI - Distinguishing heroes from celebrities. AB - Although numerous studies have concerned both 'greatness' and 'celebrity worship,' little previous research has overtly investigated the psychological bases for distinguishing 'heroes' from 'celebrities'. Study 1 indicated that, in response to 13 attitudinal measures, participants' responses to heroes could be summarized in terms of emotional attachment, drive for affiliation, and ordinariness, whereas the corresponding factors for celebrities were drive for affiliation, disdain, and ordinariness. Study 2 investigated how certain individuals might come to be perceived as producing 'great' outputs. Using a vignette, manipulations of 'dead versus alive,' and 'good conduct versus bad conduct' mediated perceptions of the emotional significance of a pop song. The results are used to generate some tentative theoretical explanations for the differences between heroism and celebrity. PMID- 15826324 TI - Mood state and gambling: using mobile telephones to track emotions. AB - Mobile telephones were used to collect data on the relationship between gambling and mood state from gamblers in the field. Seventeen gamblers called an interactive voice response system running on a computer before, during and after a gambling episode. Measures taken in this way included self-reports of anxiety/arousal, the amount of money gambled, whether the result was a win or loss, the amount won or lost, and the type of gambling engaged in. Other measures were taken during an initial briefing session using conventional questionnaires that included self-reports of anxiety/arousal taken in a non-gambling situation, dissociation during gambling, and a measure of degree of impairment of control. The results showed that subjective anxiety/arousal levels were significantly higher during and after gambling than during the urge to gamble or at baselines. Losing was associated with increased subjective anxiety/arousal after play, and winning was associated with a decrease in subjective anxiety/arousal. This suggests that gambling may be a cause of increased subjective anxiety/arousal, rather than functioning to relieve it. A cluster of variables associated with impaired control and subjective anxiety/arousal levels was also identified. The method of collecting data using mobile telephones appears to be a valuable development. PMID- 15826325 TI - Chance favours only the prepared mind: Incubation and the delayed effects of peer collaboration. AB - Research has shown that the beneficial effects of peer collaboration are not always apparent until some time has elapsed. Such delayed effects are not readily incorporated in current models of collaborative learning, but because they constitute incubation effects in the psychological sense of the term, they should in principle be consistent with cognitive accounts of how incubation occurs. Accordingly, the paper reports three studies which test whether, in accordance with cognitive models of incubation, the delayed effects of peer collaboration could result from: (a) the breaking over time of unhelpful mental sets, (b) the engagement in post-collaborative reflective appraisal, and (c) the experience of subsequent events that, thanks to collaboration, can be used productively. The studies involved 9- to 12-year-old children working on the factors relevant to floating and sinking. The results provide no evidence for the relevance of set breaking or reflective appraisal, but suggest strongly that peer collaboration can 'prime' children to make good use of subsequent input. It is argued that in addition to clarifying how the delayed effects occur and applying incubation models in a novel context, the results flag up issues that are common to collaborative and cognitive theory, and that neither address adequately. PMID- 15826326 TI - Gaze and arrow cueing of attention reveals individual differences along the autism spectrum as a function of target context. AB - Observing averted gaze results in a reflexive shift of attention to the gazed-at location. In two experiments, participants scoring high and low on the Autism Spectrum Quotient questionnaire (AQ; Baron-Cohen, Wheelwright, Skinner, Martin, & Clubley, 2001) observed arrow and gaze cues to investigate cueing effect magnitude as a function of the context in which peripheral targets could appear. While identical cueing effects were found with gaze and arrow cues, the more striking results concerned target stimuli. In Experiment 1, targets could appear on a peripheral face, or on scrambled face parts. Overall, greater cueing effects were found when the target appeared on a face. However, this face bias was only observed in participants with low AQ scores, whereas high AQ scorers oriented more to scrambled features. Experiment 2 found equal cueing to targets appearing on tools, as compared with tool parts. However, individual differences were again observed, where low AQ scorers showed larger cueing towards tools, while high scorers oriented more to scrambled parts, as in Experiment 1. These results support the idea that low AQ individuals orient strongly to objects attended by others. However, since the same results were found for arrow cues, this effect may generalize to all central cues to attention. High AQ scorers possessing many more autistic-like traits tended to orient more to scrambled shapes, perhaps reflecting a bias for orienting to local details. PMID- 15826327 TI - 'It's still bending': verbal suggestion and alleged psychokinetic ability. AB - Some alleged psychics appear to be able to deform metallic objects, such as keys and cutlery, by thought alone. This paper describes two studies that examined whether one aspect of these demonstrations could be created by verbal suggestion. In the first study, participants were shown a videotape in which a fake psychic placed a bent key on a table. Participants in one condition heard the fake psychic suggest that the key was continuing to bend, whilst those in the other condition did not. Participants in the suggestion condition were significantly more likely to report that the key continued to bend. These findings were replicated in the second study. In addition, participants who reported that the key continued to bend displayed a significantly higher level of confidence in their testimony than others, and were significantly less likely to recall that the fake psychic had suggested the continued bending of the key. Neither experiment revealed any differences between participants who expressed a prior belief in the paranormal compared with those who did not. The paper discusses the implications of these results for the psychology of suggestion and the assessment of eyewitness testimony for anomalous events. PMID- 15826328 TI - Maternal/paternal antenatal attachment and fourth-dimensional ultrasound technique: a preliminary report. AB - The purpose of the present research was to investigate the role of the fourth dimensional (4D) ultrasound scanning on the antenatal attachment development, in pregnant women (19-23 weeks of gestation) and their partners. A total of 44 couples were asked to complete questionnaires about maternal and paternal antenatal attachment, before the two-dimensional (2D; control group) and 4D (experimental group) ultrasound scanning and 2 weeks later. In the groups of women, our findings underline the progressive increase in antenatal attachment throughout gestation: the mean level of antenatal attachment was significantly different between the first and the second assessment. No significant differences are shown between 2D and 4D ultrasound scanning groups. We do not eliminate the possibility that ultrasound scanning practice would be a helpful and crucial means of investigation for a complete explanation of prenatal attachment (Ainsworth, Blehar, Waters, & Wall, 1978; Fivaz-Depeursinge & Corboz-Warney, 2000) and parental mental representations (Stern, 1987, 1997). PMID- 15826330 TI - An application of an extended health belief model to the prediction of breast self-examination among women with a family history of breast cancer. AB - OBJECTIVES: This study reports an application of the health belief model (HBM) to the prediction of breast self-examination (BSE) among women with a family history of breast cancer. The study also considered the influence of breast cancer worries and past behaviour. METHODS: Eight hundred and thirty-three women completed questionnaires, based on the HBM, to assess their beliefs about breast cancer and BSE. Of these women, 567 were followed-up at 9 months when BSE frequency was assessed. RESULTS: Discriminant function analysis was employed to discriminate among infrequent, appropriate and excessive BSE. Two functions were calculated which were predictive of group membership. The first function maximally discriminated between the infrequent BSE group and the other two groups, with infrequent selfexaminers reporting a greater number of self-efficacy and emotion barriers, fewer benefits and less frequent BSE at Time 1. The second function maximally discriminated between the excessive BSE group and the appropriate BSE group, with excessive selfexaminers reporting higher levels of breast cancer worries and perceived severity and fewer self-efficacy barriers. CONCLUSIONS: The results highlight the importance of focusing on excessive as well as infrequent BSE. Interventions designed to enhance women's confidence in their ability to perform BSE, coupled with attempts to reduce breast cancer worries, may encourage more appropriate and effective BSE. PMID- 15826331 TI - Intense-personal celebrity worship and body image: evidence of a link among female adolescents. AB - OBJECTIVES: The aim of the present study was to examine the relationship between celebrity worship and body image within the theoretical perspective of intense para-social relationships with celebrities. DESIGN: Correlation and multiple regression analyses were used to examine the relationships between celebrity worship and body image. METHOD: Three samples, 229 (102 males and 127 females) adolescents, 183 (88 males and 95 females) full-time university undergraduate students, and 289 (126 males and 163 females) adults were administered an amended version of the Celebrity Attitude Scale, the Attention to Body Shape Scale, and the Body Shape Questionnaire-Revised. RESULTS: Significant relationships were found between attitudes toward celebrities and body image only among female adolescents. Multiple regression analyses suggested that Intense-personal celebrity worship accounted for unique variance in scores in body image. CONCLUSIONS: Findings suggest that in female adolescents, there is an interaction between Intense-personal celebrity worship and body image between the ages of 14 and 16 years, and some tentative evidence has been found to suggest that this relationship disappears at the onset of adulthood, 17 to 20 years. Results are consistent with those authors who stress the importance of the formation of para social relationships with media figures, and suggest that para-social relationships with celebrities perceived as having a good body shape may lead to a poor body image in female adolescents. PMID- 15826332 TI - Evidence for controlled heroin use? Low levels of negative health and social outcomes among non-treatment heroin users in Glasgow (Scotland). AB - OBJECTIVES: This longitudinal study focused on 126 long-term heroin users who had never been in specialist treatment for use of any drug. The primary aim of the study was to assess whether this 'hidden' population resembled heroin users identified with drug treatment agencies, or alternatively, to test whether heroin could indeed be used in a controlled, non-intrusive fashion for an extended period of time. DESIGN AND METHODS: Recruitment was achieved through chain referred purposive sampling methods, and data were collected through two semi structured interviews. 67% of participants were re-recruited for follow-up. RESULTS: Participants had levels of occupational status and educational achievement comparable to that in the general UK population, and considerably higher than typically found in heroin research. At the conclusion of the study, six participants had entered treatment. While there was evidence of intensive risky patterns of drug use among the sample, there was equal evidence for planned, controlled patterns of use. Some drugrelated negative health and social outcomes had occurred on a lifetime basis, but ongoing problems were rare, and heroin was not a significant predictor in either context. In contrast to typical samples of heroin users, high levels of negative health and social outcomes did not appear to be inevitable within this sample. Frequency of heroin use was predicted by attributional items, indicating the importance of psychological factors in drug use and addiction. CONCLUSIONS: Drug research should more fully incorporate previously hidden populations to more fully inform theory and practice. The pharmacological properties of specific substances should not be assumed to inevitably lead to addictive and destructive patterns of drug use. PMID- 15826333 TI - The nature of imagery processes underlying food cravings. AB - OBJECTIVE: The study used a working memory approach to examine the nature of the imagery processes underlying food cravings. DESIGN AND METHOD: A sample of 60 dieters and 60 non-dieters were asked to imagine either a food induction or a holiday induction scenario. Participants then performed 18 trials of either a visual imagery task (loading the visuo-spatial sketch pad) or auditory imagery task (loading the phonological loop). Food craving was measured before and after the induction scenario, and then after every 6 trials of the imagery task. RESULTS: Craving intensity increased following instructions to imagine the food (but not holiday) induction scenario, especially for dieters. As predicted, the visual imagery task was superior to the auditory imagery task in reducing the level of food craving. DISCUSSION: The results confirmed the imaginal basis of food cravings. Specifically, they demonstrated that the imagery processes involved in food cravings are predominantly visual in nature. Hence concurrent tasks that load the visuospatial sketch pad of working memory can be used to reduce food cravings. The findings have potential application in the treatment of craving episodes in clinical populations. PMID- 15826334 TI - Optimism, coping style and emotional well-being in cardiac patients. AB - OBJECTIVE: Optimism is associated with superior emotional well-being in people with chronic and acute health problems, possibly because optimists are more likely to implement problem-focused coping. Another interpretation posits that optimism can be a defensive response designed to diminish affective reactions to health problems. The study objective is to investigate this possibility. DESIGN: A cross-sectional examination of relationships between dispositional and relative optimism, threat avoidance and emotional well-being in 85 cardiac patients. RESULTS: Blunting, a measure of threat avoidance, was found to be associated with both optimism and emotional well-being, and the common variance was predictive of positive affect. As expected, this link was stronger in people with low self efficacy for problem-focused coping. CONCLUSION: These findings support a defensive interpretation of optimism amongst patients with recently-experienced cardiac disease, particularly as the effect was more pronounced in the low self efficacy subsample. We discuss possible explanations for these findings and implications for the study of coping with serious illness. PMID- 15826335 TI - Two Black men with prostate cancer: a narrative approach. AB - OBJECTIVES: This paper demonstrates the value of a narrative approach for health psychology. It focuses on the lives of two Black men with prostate cancer, drawn from a larger study investigating the links between masculinities and prostate cancer. DESIGN: The study was a qualitative, interview-based study. Each participant was interviewed four times. METHODS AND ANALYSIS: The men were asked to describe and discuss their prostate cancer experience, as well as their lives prior to illness. In order to gain a perspective on individual experiences of men with prostate cancer, we took a narrative approach to gathering and analysing data. Results are reported through two descriptive narratives. CONCLUSIONS: The narratives of the men described in the paper show how the interaction of race with health and illness is neither predictable nor consistent at the individual level. Black men, like all men with prostate cancer, have diverse experiences and are influenced by a wide array of personal and societal factors. While the high risk of prostate cancer among Black men makes proactive interventions advisable, such interventions will be most effective if the heterogeneity of men's experiences are taken into account. PMID- 15826336 TI - Determinants of condom use among a random sample of single heterosexual adults. AB - OBJECTIVES: The aim of this study was to identify the determinants of single heterosexuals' use of condoms during each sexual intercourse. DESIGN: Cohort of individuals followed over a period of 2 years. METHODS: Respondents were 574 single heterosexual individuals who answered questions during a telephone interview. RESULTS: Attitude, perceived behavioural control, self-efficacy and moral norm explained 65% of the variance in intentions to use condoms ( p, .0001). The determinants of condom use at 1 year follow-up (27% explained variance) were perceived behavioural control, past behaviour, and the interaction between intention and intention stability. At 2 years follow-up, the significant determinants were past behaviour and intention by intention stability. In the present study, intentions to use condoms which remained stable across time were strong predictors of condom use (at 1 year: beta = .42, p < .0001; at 2 years: beta = .39, p, < .003), while unstable intentions were not (at 1 year: beta = .02, ns; at 2 years: beta = .19, ns). CONCLUSION: Intention stability is a major determinant of the accuracy of intention for the prediction of subsequent behaviour. PMID- 15826337 TI - Trying to do my best as a mother: decision-making in families of children undergoing elective surgical treatment for short stature. AB - OBJECTIVES: To explore how families make decisions about elective leg-lengthening surgical treatment. DESIGN: Interviews were conducted and analysed using Interpretative Phenomeno logical Analysis (Smith, 1995). METHODS: Data were gathered using semi-structured interviews with nine mothers of children who had recently decided to undergo treatment. RESULTS: Overall, the decision process was guided by the mothers' central concern to act responsibly as a parent. Thematic analysis indicated that the decision was taken in a social context where short stature could lead to discrimination and disability. The decision-making process evolved gradually over several years as mothers and children gathered information about treatment. While mothers emphasized that ultimately it was their child's decision, they monitored the decision process and filtered the information available in an attempt to ensure that the child made a well-informed and wise choice. CONCLUSIONS: The decision was presented as an ongoing process by the mothers, their concerns representing their desire to do their best as parents for their children. Theoretically, the mothers' description of the process can be understood in terms of their attempts to resolve an ethical dilemma. Clinical implications include recognition of the role of the psychologist in supporting mothers in their decision making and thus indirectly helping children. PMID- 15826338 TI - The impact of job strain on the predictive validity of the theory of planned behaviour: an investigation of exercise and healthy eating. AB - OBJECTIVES: This study examined the impact of the job strain model on exercise and healthy eating within the framework of the Theory of Planned Behaviour. DESIGN: Participants completed a questionnaire measuring the components of the Theory of Planned Behaviour and the job strain model. A follow-up questionnaire a week later measured behaviour. METHOD: The questionnaires were completed by e mail. The initial questionnaire was completed by 331 employees, and 286 follow-up questionnaires were returned. RESULTS: Job demands affected exercise indirectly by lowering perceptions of behavioural control over exercise. However, variables from the job strain model were not related to exercise intentions or behaviour. In contrast, employees in passive jobs intended to eat more sweets and snack foods and employees in low strain jobs were more likely to realize their intentions to eat more sweets and snack foods. However, variables from the job strain model did not influence consumption of fruit and vegetables. CONCLUSIONS: While people may consider the impact of job strain on exercise whilst making decisions about whether or not to exercise, job strain has a more direct impact on healthy eating. However, job strain may only affect consumption of so-called 'high density' foods, rather than foods such as fruit and vegetables. PMID- 15826339 TI - Women's opinions about attending for breast cancer screening: stability of cognitive determinants during three rounds of screening. AB - OBJECTIVES: To examine the stability of beliefs and intentions towards repeat attendance at breast cancer screening, using the Theory of Planned Behaviour. The aims of the study were to examine whether and how cognitions changed in the course of the programme, and whether intentions that were assessed proximally were better predictors of behaviour than those assessed distally. DESIGN AND METHODS: A total of 2,657 women filled out a baseline questionnaire (T(1)), 2 months after being invited for an initial mammogram in the Dutch Breast Cancer Screening Programme. Actual attendance data in the second and third screening rounds were subsequently collected and follow-up questionnaires were sent to parts of the sample at four points in time: shortly before (T(2)) and after (T(3)) the second screening round, and shortly before (T(4)) and after (T(5)) the third screening round. RESULTS: Only minor changes in beliefs and intentions were found. In the assessments shortly before screening, women were somewhat less positive about attending than in the assessments shortly after screening. Throughout the course of the programme, women's opinions about attending remained positive. In fact, women became somewhat more convinced that they were vulnerable to getting breast cancer, and that participating in screening was beneficial to them. Actual attendance in subsequent rounds of screening was higher than expected. Proximal beliefs and intentions were only slightly more predictive of actual behaviour than distal beliefs. CONCLUSIONS: In organized breast cancer screening, beliefs and intentions remain positive and rather stable. Although our results should be interpreted with caution, due to little variation in behaviour, they suggest that the gap between intentions and behaviour could not be substantially reduced by proximal assessment of determinants. PMID- 15826340 TI - Thought suppression and cognitive vulnerability to depression. AB - OBJECTIVES: Cognitive reactivity (CR) has been defined as the relative ease with which maladaptive cognitions or cognitive styles are triggered by mild (nonpathological) mood fluctuations. CR has been found to predict relapse of depression (Segal, Gemar and Williams, 1999). This study compared different measures of CR, and also investigated the role of thought suppression as a possible mechanism underlying CR. DESIGN AND METHODS: Participants included 24 previously depressed, and 24 never depressed individuals who underwent a mood induction. They also completed a questionnaire designed to measure CR (LEIDS; Van der Does, 2002a), and participated in the scrambled sentences task (SST). The SST was designed to uncover thought suppression tendencies, and has been shown to discriminate between never depressed and previously depressed samples. RESULTS: LEIDS scores were higher for previously depressed than for never depressed individuals. However, CR as measured with the mood induction did not distinguish between these groups. The LEIDS was correlated with the results of the SST and with self-report measures of thought suppression. CONCLUSION: Active suppression of unwanted thoughts may be involved in the apparent inactive state of depressive cognitions during remission. PMID- 15826341 TI - Are measures of hypomanic personality, impulsive nonconformity and rigidity predictors of bipolar symptoms? AB - OBJECTIVES: It has been suggested that temperaments such as 'hypomanic personality' (HYP) have an explanatory role in affective disorders. Similarly, the impulsive nonconformity scale, originally designed to assess psychosis proneness, was recently found to augment the prediction of manic episodes. Conversely, research indicates that 'rigidity', a central feature of Typus Melancholicus (TMEL; von Zerssen, 1996), may characterize the premorbid personality of depressives. DESIGN: The present study combines these three scales to prospectively predict manic and depressive symptoms diagnosed 2 years later in a non-college student population. METHOD: Structured clinical interviews for DSM IV were conducted with 114 individuals (60% female, mean age = 19 . 9 years), 2 years after an initial screening. RESULTS: It was found that none of the predictors predicted purely depressive symptoms. As expected, HYP emerged as the strongest predictor of (hypo-)manic symptoms. CONCLUSIONS: While rigidity did not predict depression, people with a hypomanic temperament are at risk of developing symptoms of bipolar disorder, especially (hypo-)manic ones. It is noteworthy that this was evident in our young sample, still in the earlier stages of the high risk period. PMID- 15826342 TI - Sentence completion test for depression (SCD): an idiographic measure of depressive thinking. AB - OBJECTIVES: This study set out to investigate the reliability and validity of the Sentence Completion Test for Depression (SCD) as a clinical measure. In contrast to questionnaire measures of depressive thinking, respondents finish incomplete sentences using their own words. This elicits idiographic information concurrent with measuring depressive thinking. METHOD: In Study 1, measures of negative thinking were tested between a depressed group and a non-depressed control group. A preliminary item analysis was conducted and replicated on separate samples in Study 2. Psychometric properties of the test were investigated. In Study 3, idiographic validity and sensitivity to change were explored in a sample of clinical cases with reference to cognitive-behavioural case-formulation. RESULTS: In Study 1, the depressed group produced more negatives and fewer positives, and the SCD demonstrated good content validity, internal consistency and inter-rater reliability. The preliminary short-form had comparable psychometric properties, and these were replicated on new samples in Study 2. Sensitivity and specificity values were above 90% in both studies. In Study 3, idiographic content generated hypotheses about target problems and dysfunctional beliefs within cognitive behavioural case-formulation, and SCD scores were sensitive to clinical change. CONCLUSIONS: The SCD demonstrates good construct validity, internal consistency, inter-rater reliability, sensitivity, and specificity. It offers an idiographic assessment of depression that is complementary to questionnaire measures, particularly by generating hypotheses about target problems and dysfunctional beliefs within a cognitive-behavioural case-formulation. This is achieved without loss to reliability and validity at the nomothetic level. PMID- 15826343 TI - Neuropsychological correlates of the YAQ-S and YAQ-I self- and informant-reported ADHD symptomatology, emotional and social problems and delinquent behaviour. AB - OBJECTIVES: The objective was to evaluate the relationship between self- and informant-rated symptoms of ADHD and performance on neuropsychological tests of attention and impulsivity (Letter Cancellation Test [LCT], Continuous Performance Test [CPT] and Matching Familiar Figures test [MFF]). DESIGN: Participants were clinical referrals for assessment of ADHD in adulthood and healthy controls. METHOD: ADHD adults were compared with individuals with mild psychiatric disorders (including attention problems), a healthy control group on neuropsychological measures of attention (CPT and LCT) and on impulsivity (MFF). Secondly, the scores obtained on these measures were correlated with the Young ADHD Questionnaire-S (YAQ-S) and Young ADHD Questionnaire-I (YAQ-I) self- and informant-reported scales of ADHD symptomatology, emotional problems, delinquency, and social functioning. RESULTS: The ADHD group were more impaired than the healthy controls on neuropsychological measures, but only the MFF differentiated them from the psychiatric controls. Tests of attentional control positively correlated with the selfreported functioning of the ADHD group but not among the psychiatric or normal controls. There was a positive correlation between tests of attention (but not impulsivity) and self-ratings of ADHD symptomatology for the ADHD group, suggesting that ADHD adults are better at recognizing attentional problems than impulse control. A positive correlation was found between the Delinquency scale and a test of impulsivity for the ADHD group only. CONCLUSIONS: The findings suggest that there are neuropsychological underpinnings of the ADHD syndrome but neuropsychological tests may discriminate poorly between ADHD and mild psychiatric disorders. ADHD adults may be more reliable informants of adult functioning than their parents, although they may underreport impulsive behaviour. PMID- 15826344 TI - The dilemmas of residential care staff working with the challenging behaviour of people with learning disabilities. AB - OBJECTIVES: To investigate residential care staff beliefs and feelings about the challenging behaviour of adults with learning disabilities in their care, and how they perceive these beliefs and feelings to have developed over time. DESIGN: A qualitative study using thematic analysis. METHODS: A group of 18 staff from 10 different residential services participated in indepth semi-structured interviews. Transcripts were analysed according to thematic analysis techniques drawn from interpretative phenomenological analysis (IPA) and grounded theory. The analysis was then subjected to scrutiny by participants using a respondent validation survey. RESULTS: Staff talked of dilemmas about whether challenging behaviour should be seen as a 'communication' of need or as a 'behaviour problem', how to balance a 'firm' response with 'kindness', and how to deal with unpleasant feelings evoked by the work, especially fear and frustration. Over time, staff reported overcoming initial fears of the client by 'getting to know them', or alternatively, avoiding the client, 'cutting off' emotionally, or protecting themselves with safety procedures. CONCLUSIONS: The analysis suggests that staffs are troubled by the limitations of a narrow behavioural discourse. Staff development and training based on richer approaches that integrate behavioural ideas with a value-based philosophy, might allow staff to respond to needs expressed by behaviour without fear of reinforcing it. Services should attend to staff emotional reactions to their work, to help them maintain nonavoidant coping strategies. PMID- 15826345 TI - Visual hallucinations in psychiatric conditions: appraisals and their relationship to distress. AB - OBJECTIVES: Visual hallucinations (visions) associated with psychiatric conditions are poorly understood. This study tested the hypothesis that appraisals of visions would predict distress associated with visions. It also aimed to describe typical appraisals of visions. DESIGN: A cross-sectional interview study of people with visual hallucinations. METHOD: Participants (N = 20) who had experienced visions caused by their psychiatric condition (i.e. not by additional pathology such as epilepsy or substance abuse) completed a semi structured interview and the Beck Depression Inventory (BDI). Possible associations between appraisals of visions (e.g. of controllability or outcome) and distress were examined. Novel categories of appraisal were generated in an attempt to reflect individuals' experiences as closely as possible. RESULTS: Appraisals of negative outcome, but not control, predicted distress. Thus, if individuals believed that the persistence of their visions would result in unpleasant consequences, they experienced more distress. Neither the form nor content of the visions predicted distress, nor was current mood predictive. Five categories of appraisal were generated, and independent raters could assign these reliably. Preliminary analyses showed that where individuals appraised a vision as indicating a special task, gifts or powers, they experienced less distress. CONCLUSIONS: Appraisals of visions predicted the distress associated with visions. 'Objective' characteristics of visions were poorer predictors of distress, and appraisals did not simply follow from the content of the vision. However, current mood may partially determine appraisals. The novel categories of distress require replication but may help to guide clinical practice. Therapeutic interventions aimed at altering appraisals may reduce distress. PMID- 15826346 TI - Early separation anxiety and adult attachment style in women with eating disorders. AB - OBJECTIVE: To ascertain whether women with eating disorders have a higher frequency of separation anxiety symptoms in childhood, and a higher prevalence of insecure styles of adult attachment, compared with healthy women. METHODS: The Separation Anxiety Symptom Inventory (SASI) and the Attachment Style Questionnaire (ASQ) were administered to 78 women with eating disorders and 64 healthy women. RESULTS: Compared with control women, women with eating disorders reported more severe symptoms of separation anxiety during childhood, and scored higher on the ASQ scales, reflecting insecure styles of adult attachment. Early separation anxiety and insecure attachment were not correlated with age of onset or illness duration. The diagnostic subgroup was not associated with a specific style of insecure attachment: compared with control women, both anorexic and bulimic women scored higher on the ASQ scales reflecting anxious attachment, but not on the scales reflecting avoidant attachment. CONCLUSIONS: The results confirm the link between eating disorders and insecure attachment that has been found in previous studies, and extend it to childhood symptoms of separation anxiety. PMID- 15826347 TI - Coping with depression and vulnerability to mania: a factor analytic study of the Nolen-Hoeksema (1991) Response Styles Questionnaire. AB - OBJECTIVES: To assess the factor structure of the Nolen-Hoeksema (1991) Response Styles Questionnaire (RSQ), and to investigate the relationship between coping with depression and other measures of affective symptomatology in a student sample. DESIGN: A factor analytic study of the RSQ followed by an investigation of the relationship between RSQ scale scores and measures of affective symptomatology. METHOD: Five hundred twenty-eight undergraduate students completed a battery of questionnaires comprising the RSQ, Beck Depression Inventory, Hypomania Personality Questionnaire, Positive and Negative Affect Scale and the Dysfunctional Attitudes Scale. Factor analysis of the RSQ was performed and correlational and regression analyses were conducted on the data from the other four questionnaire measures. RESULTS: Factor analysis revealed a meaningful 3-factor solution which measured coping styles characterized by (i) rumination; (ii) pleasant distraction and problemsolving; and (iii) risk-taking. Rumination and risk-taking were independently associated with both depression and hypomania scores, and hypomania was associated with both depression and dysfunctional attitudes. Additional relationships between coping and the other measures are also reported. CONCLUSIONS: The results support the validity of the coping styles concept and suggest their natural separation into three distinct strategies. Abnormal coping is discussed as a potential contributing factor to affective symptoms, including symptoms of bipolar disorder. PMID- 15826348 TI - Development of depressed mood predicts onset of psychotic disorder in individuals who report hallucinatory experiences. AB - OBJECTIVES: Current psychological theories state that the clinical outcome of hallucinatory experiences is dependent on the degree of associated distress, anxiety, and depression. This study examined the hypothesis that the risk for onset of psychotic disorder in individuals with self-reported hallucinatory experiences would be higher in those who subsequently developed depressed mood than in those who did not. DESIGN: A prospective cohort study of a general population sample. METHODS: A sample of 4,670 individuals with no lifetime evidence of any psychotic disorder were interviewed with the Composite International Diagnostic Interview Schedule (CIDI) at baseline and 1 and 3 years later. At Year 3, individuals with CIDI evidence of psychotic symptoms were interviewed by clinicians to identify potential onset of psychotic disorder. Psychotic disorder was specified at three levels; two involving severity of positive symptoms of psychosis, and one using additional clinical judgment of need for care. RESULTS: Given the presence of hallucinatory experiences at baseline, the increase in risk of having the psychosis outcome at Year 3 was higher in the group with depressed mood at Year 1 than in the group without depressed mood at Year 1 (any level of psychotic symptoms: risk difference 17.0%, 95% CI - 1.7, 35.7; severe level of psychotic symptoms: risk difference 21.7%, 95% CI 3.2, 40.2; needs-based diagnosis of psychotic disorder: risk difference 16.8%, 95% CI 0.4, 33.3). CONCLUSION: The results are in line with current psychological models of psychosis that emphasize the role of secondary appraisals of psychotic experiences in the onset of clinical disorder. PMID- 15826349 TI - The prediction of hallucinatory predisposition in non-clinical individuals: examining the contribution of emotion and reasoning. AB - BACKGROUND: Emotion, especially anxiety, has been implicated in triggering hallucinations. Reasoning processes are also likely to influence the judgments that lead to hallucinatory experiences. We report an investigation of the prediction of hallucinatory predisposition by emotion and associated processes (anxiety, depression, stress, self-focused attention) and reasoning (need for closure, extreme responding). METHOD: Data were analysed from a questionnaire survey in a student population (N = 327). RESULTS: Higher levels of anxiety, self focus, and extreme responding were associated with hallucinatory predisposition. Interactions between these three variables did not strengthen the predictive effect of each. Depression, stress, and need for closure were not found to be predictors of hallucinatory experience in the regression analysis. CONCLUSION: Emotional and reasoning processes may both need to be considered in the understanding of hallucinatory experience. PMID- 15826350 TI - Autobiographical memory specificity and emotional abuse. AB - OBJECTIVE: Examined the relationship between trauma and memory specificity and the importance in this of level of support received. METHOD: Fifty-two female undergraduates completed the Autobiographical Memory Test (AMT; Williams and Broadbent, 1986), assessing memory specificity and the Traumatic Experiences Checklist (TEC; Nijenhuis, Van der Hart, and Vanderlinden, 1999), assessing qualitative aspects of trauma. RESULTS: In low-specific students, those who reported emotional abuse were less specific in their memory. The scores of students who had not received any support for the abuse showed a trend towards retrieval of less specific memories, as compared with those who had received support. CONCLUSION: Results provide evidence for an association between trauma (emotional abuse) and reduced memory specificity, and suggest that receiving support following the abuse might protect individuals from developing over general memory. PMID- 15826353 TI - The many faces of sequence alignment. AB - Starting with the sequencing of the mouse genome in 2002, we have entered a period where the main focus of genomics will be to compare multiple genomes in order to learn about human biology and evolution at the DNA level. Alignment methods are the main computational component of this endeavour. This short review aims to summarise the current status of research in alignments, emphasising large scale genomic comparisons and suggesting possible directions that will be explored in the near future. PMID- 15826354 TI - Bioinformatics analysis of alternative splicing. AB - Over the past few years, the analysis of alternative splicing using bioinformatics has emerged as an important new field, and has significantly changed our view of genome function. One exciting front has been the analysis of microarray data to measure alternative splicing genome-wide. Pioneering studies of both human and mouse data have produced algorithms for discerning evidence of alternative splicing and clustering genes and samples by their alternative splicing patterns. Moreover, these data indicate the presence of alternative splice forms in up to 80 per cent of human genes. Comparative genomics studies in both mammals and insects have demonstrated that alternative splicing can in some cases be predicted directly from comparisons of genome sequences, based on heightened sequence conservation and exon length. Such studies have also provided new insights into the connection between alternative splicing and a variety of evolutionary processes such as Alu-based exonisation, exon creation and loss. A number of groups have used a combination of bioinformatics, comparative genomics and experimental validation to identify new motifs for splice regulatory factors, analyse the balance of factors that regulate alternative splicing, and propose a new mechanism for regulation based on the interaction of alternative splicing and nonsense-mediated decay. Bioinformatics studies of the functional impact of alternative splicing have revealed a wide range of regulatory mechanisms, from NAGNAG sites that add a single amino acid; to short peptide segments that can play surprisingly complex roles in switching protein conformation and function (as in the Piccolo C2A domain); to events that entirely remove a specific protein interaction domain or membrane anchoring domain. Common to many bioinformatics studies is a new emphasis on graph representations of alternative splicing structures, which have many advantages for analysis. PMID- 15826355 TI - Putting microarrays in a context: integrated analysis of diverse biological data. AB - In recent years, multiple types of high-throughput functional genomic data that facilitate rapid functional annotation of sequenced genomes have become available. Gene expression microarrays are the most commonly available source of such data. However, genomic data often sacrifice specificity for scale, yielding very large quantities of relatively lower-quality data than traditional experimental methods. Thus sophisticated analysis methods are necessary to make accurate functional interpretation of these large-scale data sets. This review presents an overview of recently developed methods that integrate the analysis of microarray data with sequence, interaction, localisation and literature data, and further outlines current challenges in the field. The focus of this review is on the use of such methods for gene function prediction, understanding of protein regulation and modelling of biological networks. PMID- 15826356 TI - Bioinformatics approaches and resources for single nucleotide polymorphism functional analysis. AB - Since the initial sequencing of the human genome, many projects are underway to understand the effects of genetic variation between individuals. Predicting and understanding the downstream effects of genetic variation using computational methods are becoming increasingly important for single nucleotide polymorphism (SNP) selection in genetics studies and understanding the molecular basis of disease. According to the NIH, there are now more than four million validated SNPs in the human genome. The volume of known genetic variations lends itself well to an informatics approach. Bioinformaticians have become very good at functional inference methods derived from functional and structural genomics. This review will present a broad overview of the tools and resources available to collect and understand functional variation from the perspective of structure, expression, evolution and phenotype. Additionally, public resources available for SNP identification and characterisation are summarised. PMID- 15826357 TI - A survey of current work in biomedical text mining. AB - The volume of published biomedical research, and therefore the underlying biomedical knowledge base, is expanding at an increasing rate. Among the tools that can aid researchers in coping with this information overload are text mining and knowledge extraction. Significant progress has been made in applying text mining to named entity recognition, text classification, terminology extraction, relationship extraction and hypothesis generation. Several research groups are constructing integrated flexible text-mining systems intended for multiple uses. The major challenge of biomedical text mining over the next 5-10 years is to make these systems useful to biomedical researchers. This will require enhanced access to full text, better understanding of the feature space of biomedical literature, better methods for measuring the usefulness of systems to users, and continued cooperation with the biomedical research community to ensure that their needs are addressed. PMID- 15826358 TI - Current efforts in the analysis of RNAi and RNAi target genes. AB - RNAi is RNA interference by short RNAs. It influences gene-expression by down regulation of mRNAs, typically by complementarity to the 3' UTR (untranslated region) of the mRNA. microRNAs (miRNAs) are short RNAs acting as natural RNAi. miRNAs mediate down-regulation of many mRNAs from developmental genes and transcription factor genes. Natural examples for this additional level of post transcriptional control are increasing. Suitable computer-based search strategies for new miRNA candidates include precursor folding as well as different compositional search strategies. Example programs for this are presented. New own and other data are provided for an overview on such strategies. A strategy feasible in plants for miRNA target identification is direct base pairing of miRNAs to potential mRNA target 3' UTRs. Correct identification in animals usually requires comparative genomics and conserved UTR regions pairing to conserved miRNA substructures. A number of example programs and target examples for these tasks are examined. Finally, strategies and programs for artificial gene silencing by designed RNAi are explained. PMID- 15826359 TI - There is no silver bullet--a guide to low-level data transforms and normalisation methods for microarray data. AB - To overcome random experimental variation, even for simple screens, data from multiple microarrays have to be combined. There are, however, systematic differences between arrays, and any bias remaining after experimental measures to ensure consistency needs to be controlled for. It is often difficult to make the right choice of data transformation and normalisation methods to achieve this end. In this tutorial paper we review the problem and a selection of solutions, explaining the basic principles behind normalisation procedures and providing guidance for their application. PMID- 15826361 TI - Importance of Debye and Keesom interactions in separating m-xylene and p-xylene in GC-MS analysis utilizing PEG stationary phase. AB - In electron impact gas chromatography (GC)-mass spectrometry analysis of a complex mixture, such as gasoline, two coeluting solutes can be distinguished if each has a unique major ion. The boiling-point difference between m-xylene and p xylene, which also has identical major ions (mz-1: 91 and 106 Da), is 0.77 degrees C. These cannot be separated even on a crossed-linked polydimethylsiloxane capillary column, which has a minimum of 5000 plates/m. They are separated on a crossed-linked polar polyethylene glycol (PEG) capillary column. GC separation on a stationary phase depends on the relative strengths of solute-solute, stationary phase-stationary phase, and solute-stationary phase interactions. Although the calculated molar electronic polarization and refractivity factor of Lorenz-Lorentz equation for m-xylene and p-xylene are nearly equal because of its greater dipole moment difference (0.30 and 0.02), the calculated orientation polarization of m-xylene is 80 times greater than p xylene. This implies the dipole reinforcement through inductive polarization by the hydroxyl of PEG stationary phase molecules is greater on m-xylene than p xylene. In addition, as the permanent dipole moment of m-xylene is 15 times greater than p-xylene, m-xylene has a stronger Keesom interaction with PEG. In order for m-xylene and p-xylene to solvate in PEG, analytes must overcome the PEG PEG Keesom/hydrogen bonding interaction forces. Physical and chemical parameters indicate that compared with p-xylene, m-xylene has a greater Debye-Keesom interaction tendency with PEG molecules. This is supported by the 0.12-min. retention-time difference between them. PMID- 15826362 TI - Improvement of doxazosin determination in human plasma using high-performance liquid chromatography with fluorescence detection. AB - A simple, sensitive, rapid, and reproducible high-performance liquid chromatographic method is developed and validated for the determination of doxazosin in human plasma without a solvent extraction procedure. This method involves plasma protein precipitation using methanol. The structurally related compound prazosin is used as an internal standard. Doxazosin is detected with high sensitivity using spectrofluorimetry. Over the concentration range 0.5-20 ng/mL, the absolute recovery values are all greater than 98%. The method has a quantitation limit of 0.5 ng/mL. The intra- and interday coefficient of variation and inaccuracy values are all less than 8% and 7%, respectively. Therefore, the method has been applied in pharmacokinetic studies of doxazosin. PMID- 15826363 TI - Plate number requirements for establishing method suitability. AB - Establishing the suitability of an analytical system has become a routine requirement in the testing of modern pharmaceuticals. Acceptable parameters that illustrate the system is performing as intended and in an equivalent manner to the original validation are often set at the time of method validation and transferred with the method to the production laboratory. For chromatographic methods, these parameters include--but are not limited to--resolution, tailing, and plate number specifications. Transferring methods is often a seamless transition from research to quality control. However, far too often the quality group receives arguably "overzealous" and strict requirements for the method. More specifically, chromatographic methods get issued with plate number specifications that far exceed the minimum number required to achieve sufficient resolution of the analytes. Presented here is a discussion of the setting of realistic plate number specifications that still maintain the minimum resolution of the chromatographic critical pair. PMID- 15826364 TI - HPTLC method for the simultaneous determination of amlodipine and benazepril in their formulations. AB - A new, simple, precise, rapid, and selective high-performance thin-layer chromatographic (HPTLC) method is developed for the simultaneous analysis of amlodipine and benazepril in pharmaceutical formulations. The method uses zolpidem as an internal standard (IS). The stationary phase used is silica gel 60 F254 prewashed with methanol. The mobile phase consists of an ethyl acetate methanol-ammonia solution (8.5:2.0:1.0, v/v/v). Detection and quantitation are performed densitometrically at lambda = 254 nm. The Rf values of amlodipine, benazepril, and zolpidem (IS) are 0.58, 0.50, and 0.78, respectively. The limits of detection of amlodipine and benazepril are 0.02 and 0.2 microg; linearity ranges are 0.1-0.8 and 0.2-2.0 microg; and the percentage recoveries are 99.79% and 100.25%, respectively. PMID- 15826365 TI - Trace analysis of ethinyl estradiol in casein diet using gas chromatography with electron capture detection. AB - Ethinyl estradiol (EE2) is an extremely potent synthetic estrogen and a common component in oral contraceptives. The drug has a well-characterized pharmacological profile and is used as a positive control in toxicological investigations of compounds having estrogenic activity. An analytical method developed for the determination of low microg/kg levels of EE2 in a casein-based rodent diet is presented. A methanol extract of casein diet is purified for instrumental analysis by a 3-fold solid-phase extraction process. The sample extract is derivatized with pentafluoropropionic anhydride to the pentafluoropropionyl product and analyzed by capillary gas chromatography with electron-capture detection. Recoveries of EE2 from casein diet fortified at 5, 10, and 50 microg/kg average 88.8% and have a relative standard deviation (%) of 7.2. The method limit of detection in a casein-based diet is 1 microg/kg. PMID- 15826366 TI - Rapid analysis of 2,4-D in soil samples by modified Soxhlet apparatus using HPLC with UV detection. AB - The 2,4-dichlorophenoxy acetic acid (2,4-D) is used as a systemic herbicide to control broadleaf weeds in wheat, corn, range land/pasture land, sorghum, and barley. In this study, a fast and efficient method is developed by selection of modified extraction apparatus and high-performance liquid chromatography (HPLC) UV conditions for the determination of 2,4-D in soil samples. The method is applied to the study of soil samples collected from the agricultural field. The herbicide is extracted from soil samples by acetonitrile in a modified Soxhlet apparatus. The advantages of the apparatus are that it uses small volume of organic solvent, reduced time of extraction, and better recovery of the analyte. The extract is filtered using a very fine microfiber paper. The total extract is concentrated in a rotatory evaporator, dried under ultrahigh pure N2, and finally reconstituted in 1 mL of acetonitrile. HPLC-UV at 228 nm is used for analysis. The herbicide is identified and quantitated using the HPLC system. The method is validated by the analysis of spiked soil samples. Recoveries obtained varied from 85% to 100% for spiked soil samples. The limit of quantitation (LOQ) and the limit of detection (LOD) are 0.010 and 0.005 parts per million (ppm), respectively, for spiked soil samples. The LOQ and LOD are 0.006 and 0.003 ppm for unspiked soil samples. The measured concentrations of 2,4-D in spiked soil samples are between 0.010 and 0.020 ppm with an average of 0.016 +/- 0.003 ppm. For unspiked soil samples it is between 0.006 ppm and 0.012 ppm with an average of 0.009 +/- 0.002 ppm. The measured concentrations of 2,4-D in soil samples are generally low and do not exceed the regulatory agencies guidelines. PMID- 15826367 TI - Superheated water extraction of essential oils of Origanum micranthum. AB - Superheated water extraction is used to extract essential oil of leaves of Origanum micranthum. The effect of different temperatures on the essential oil profile and rate of extraction as a function of time is investigated. The components of essential oil of Origanum micranthum are removed from the aqueous extract by C18 solid-phase extraction. The identification of components is carried out using comprehensive gas chromatography-time of flight-mass spectrometry. The number of extracted components is almost the same; however, the concentrations change with changing temperature. The highest yield (0.64%) is found at a temperature of 150 degrees C, 2 mL/min and 60 bar for 30 min. The increasing temperature from 100 degrees C to 175 degrees C increased the rate of extraction of six selected components of essential oil of Origanum micranthum. cis-Sabinenehydrate exhibits the fastest rate of extraction at all temperatures studied. Some degradation products are observed at a temperature of 175 degrees C. PMID- 15826368 TI - The identification of 3,4-MDMA from its mass equivalent isomers and isobaric substances using fast LC-ESI-MS-MS. AB - 3,4-methylenedioxymethamphetamine (3,4-MDMA, "Ecstacy") and its 17 isomers and isobaric substances are studied using liquid chromatography (LC)-positive electrospray ionization-mass spectrometry (MS). 3,4-MDMA is a controlled substance, whereas in many countries the other studied isobaric compounds are not. A method for confirmation of the presence of 3,4-MDMA in drug seizures is developed and validated. Using single MS, the compounds produce an intense protonated molecule and some characteristic fragments; but tandem MS (MS-MS) is applied to enhance specificity. The MS-MS fragmentation is studied in order to distinguish 3,4-MDMA from the other 17 related compounds. However, the MS-MS spectra of 3,4-MDMA and six related compounds are very similar. Therefore, the LC MS-MS method is developed for the unambiguous identification of 3,4-MDMA. The use of a monolithic column allows for 5-min gradient runs. This qualitative method is tested with 49 Ecstacy samples seized by the police. All results are congruent with the ones obtained with other methods. PMID- 15826369 TI - Rapid speciation analysis of Cr(VI) and Cr(III) by reversed-phase high performance liquid chromatography with UV detection. AB - A simple and rapid method is developed for the simultaneous determination of Cr(VI) and Cr(III) based on the formation of their different complexes with ammonium pyrrolidine-dithiocarbamate (APDC). Separation is performed using reversed-phase high-performance liquid chromatography coupled with UV detection. The conditions for complex formation and speciation are determined, such as solution pH, amount of APDC, temperature, and type of mobile phase. In order to substantially reduce the analysis time, the separation is carried out without extraction of chromium-APDC complexes from the mother liquor. Under the optimum analysis conditions, the chromatograms obtained show good peak separation, and the absolute detection limits (3s) are 2.2 microg/L for Cr(VI) and 4.5 microg/L for Cr(III). The calibration curves are linear from 3 to 5000 microg/L for Cr(VI) and 5 to 3000 microg/L for Cr(III). The relative standard deviations of peak areas in five measurements using a sample solution of 200 microg/L are less than 2% for Cr(VI) and 4% for Cr(III), indicating good reproducibility for this analytical method. Furthermore, simultaneous determination of Cr(VI) and Cr(III) is successful with the application of the proposed procedure in the synthetic wastewaters containing common heavy metal ions: Fe(III), Pb(II), Cd(II), Cu(II), and Zn(II). PMID- 15826370 TI - Role of sulfur compounds in the detection of amino acids by ninhydrin on TLC plate. AB - Three new sulfur reagents for specific identification of amino acids on thin layer chromatography plates have been introduced. These three sulfur containing reagents are capable of developing various distinguishable colors with many of them. A probable mechanism for such color formation is proposed. PMID- 15826372 TI - The dynamics of LPS recognition: complex orchestration of multiple receptors. AB - The molecular mechanisms that have been designed to protect the host from invading pathogens are responsible for sepsis, an often fatal response of the immune system against microbial pathogens. In the past few years, intense research in the field of innate immunity has identified a plethora of pattern recognition receptors that are responsible for bacterial-induced activation. Recognition of bacterial lipopolysaccharide seems to involve a complex orchestration of protein-protein interactions that eventually leads to cellular activation. In this review, we attempt to unravel the dynamic interactions that occur among the different receptors involved and dictate the outcome of the innate immune response. PMID- 15826373 TI - Porphyromonas gingivalis fimbriae are pro-inflammatory but do not play a prominent role in the innate immune response to P. gingivalis. AB - The fimA gene encodes the major fimbrial protein of Porphyromonas gingivalis. It has been shown to stimulate adhesion to salivary proteins and other bacteria. It is also thought to play a major role in invading and stimulating host cells. To determine whether the fimA gene represents one of the principal molecules of P. gingivalis that induces inflammation, we tested purified FimA protein and a mutant P. gingivalis (DPG3) that lacks the fimA gene versus wild-type (WT) P. gingivalis. When injected into connective tissue of the scalp, purified FimA protein induced TNF-alpha and MIP-2 expression confirming that it is pro inflammatory. WT P. gingivalis induced TNF-alpha expression and recruitment of PMNs in the same model. However, DPG3 P. gingivalis stimulated TNF expression and PMN recruitment to the same extent. The latter was consistent with similar induction of the chemokine MIP-2. Similar results were obtained with diabetic mice that have a more prolonged inflammatory response to bacterial stimulation. These results indicate that FimA is a potent inducer of inflammatory cytokine expression but, in the context of P. gingivalis infection, it is not a principal stimulator of the innate host response. PMID- 15826374 TI - Lipoprotein-bound endotoxin exerts an immunomodulatory effect on hepatocytes through the lipid A domain of LPS. AB - BACKGROUND: We have previously shown that chylomicron (CM)-bound lipopolysaccharide (LPS) inhibits the host innate immune response by rendering hepatocytes tolerant to pro-inflammatory cytokine stimulation. However, LPS is a complex macromolecule containing both lipid and carbohydrate domains. We hypothesized that just as lipid A confers the toxicity of LPS, it is also responsible for the immunoregulatory effect on hepatocytes. METHODS: We pretreated primary rat hepatocytes for 2 h with a series of CM-LPS complexes in which the endotoxin moiety varied in its structure and/or toxicity. Subsequently, the cells were stimulated with a mixture of pro-inflammatory cytokines. Nitric oxide production was measured as an indicator of hepatocellular activation. RESULTS: All pretreatments wherein the CM-bound complex contained the lipid A moiety readily inhibited the hepatocellular cytokine response, including CM bound to lipid A alone. In contrast, CM-LPS complexes containing detoxified LPS, which lacks the lipid A domain, had no effect on the hepatocellular response to cytokines. CONCLUSIONS: The lipid A domain of the LPS macromolecule is both sufficient and essential for the CM-mediated induction of cytokine tolerance in hepatocytes. However, this process is independent of the specific endotoxic activity of the lipid A moiety. PMID- 15826375 TI - Optimisation of pyrogen testing in parenterals according to different pharmacopoeias by probabilistic modelling. AB - The rabbit test to detect pyrogenic contamination in parenterals is crucial to ensure patient safety. The pharmacopoeial tests in Europe, the US and Japan are based on the fever reaction of rabbits, but differ in their experimental design and in their algorithms to assess contamination. Employing an international reference endotoxin, fever can be induced in rabbits. Data from 171 rabbits built the base for probabilistic modelling of the fever reaction and for the comparison of the pharmacopoeial tests. The rabbit fever reaction could be modelled as a function of the amount of injected endotoxin (per kg body weight) by linear regression. Combining the pharmacopoeial algorithms of the rabbit pyrogen test with the developed model allowed analysis of differences regarding test results and animal consumption. This showed that the assessment of pyrogenic contamination strongly depends on the respective pyrogen test stipulated by regulations. Additionally, the approach was used to develop a new experimental design. Two specific versions of this design resulted in a reduction of the number of animals used by about 30% while the safety of the test was maintained. A need for harmonisation is evident, allowing optimisation of the experimental design, which promotes animal welfare. PMID- 15826376 TI - Role of transforming growth factor-beta1 (TGF-beta1) in endotoxin-induced hepatic failure after extensive hepatectomy in rats. AB - Postoperative infections after hepatectomy sometimes lead to fatal hepatic failure, but the mechanism of the hepatic failure is unclear. Wistar rats underwent 90% hepatectomy, and were then divided into three groups: (i) the SAL group, injected with normal saline; (ii) the LPS group, injected with lipopolysaccharide (LPS) every day for 1 week; and (iii) the LPS plus TGF-Ab (LPS+TGF-Ab) group, injected with LPS with anti-transforming growth factor-beta1 (TGF-beta1) antibody. We investigated survival rates, TGF-beta1 expression in the liver, liver regeneration by proliferating cell nuclear antigen labeling index, hepatocyte apoptosis by single stranded DNA labeling index, and perisinusoidal fibrosis using Masson's trichrome staining. The LPS group (30.4%) had a significantly lower survival rate than the SAL group (84%) and tended to be lower than the LPS+TGF-Ab group (49.4%). Liver regeneration in the LPS group was significantly lower than in the other groups. In the LPS group, hepatocyte apoptosis and perisinusoidal fibrosis was significantly more remarkable, and TGF beta1 expression was significantly higher than in the SAL group. TGF-beta1 enhanced by LPS plays an important role in the mechanism of hepatic failure by infections after hepatectomy, especially in inhibition of liver regeneration, and induction of hepatocyte apoptosis and perisinusoidal fibrosis. PMID- 15826377 TI - Crystal structure of a peptidoglycan recognition protein (PGRP) in complex with a muramyl tripeptide from Gram-positive bacteria. AB - Peptidoglycan recognition proteins (PGRPs) are pattern recognition receptors of the innate immune system that bind, and in some cases hydrolyse, bacterial peptidoglycans (PGNs). We determined the crystal structure of the C-terminal PGN binding domain of human PGRP-Ialpha in complex with a muramyl tripeptide representing the conserved core of lysine-type PGNs. The peptide stem of the ligand is buried at the deep end of a long binding groove, with N-acetylmuramic acid situated in the middle of the groove, whose shallow end could accommodate N acetylglucosamine. Both peptide and glycan moieties are essential for binding by PGRPs. Conservation of key PGN-contacting residues indicates that all PGRPs employ this basic PGN-binding mode. The structure identifies variable residues that likely mediate discrimination between lysine- and diaminopimelic acid-type PGNs. In addition, we propose a mechanism for PGN hydrolysis by Zn2+-containing catalytic PGRPs. PMID- 15826378 TI - Production and purification of recombinants of mouse MASP-2 and sMAP. AB - Small mannose-binding lectin (MBL)-associated protein (sMAP) is a component of the complex consisting of MBL and MBL-associated serine proteases (MASPs) in the lectin complement pathway. sMAP is a truncated form of MASP-2, which is generated by an alternative splicing from a single structural MASP-2 gene. Upon activation of the MBL-MASPs complex, MASP-2 cleaves the complement C4, but the role of sMAP which lacks the serine protease domain is not clear. To clarify the role of sMAP in activation of the lectin pathway, we have generated sMAP-gene deficient mice which are also deficient for MASP-2. In this study, we generated and purified mouse recombinant sMAP (rsMAP) and rMASP-2 using the Drosophila expression system for the reconstitution assay of the deficient mice. In preliminary experiments, these purified recombinants were able to reconstitute the MBL-MASPs-sMAP complexes and the addition of rMASP-2 to deficient serum restored the C4 cleavage activity of the MBL-MASPs complex. From these data, rsMAP and rMASP-2 generated in this study seem to be useful in analysis of the deficient mice. PMID- 15826379 TI - Evolution and integration of innate immune recognition systems: the Toll-like receptors. AB - Toll-like receptors (TLRs) recognize specific components of micro-organisms and trigger the activation of innate immunity. TLR-mediated signaling pathways are now rapidly being elucidated. TLR signaling originates from the cytoplasmic Toll/IL-1 receptor (TIR) domain, which is conserved among all TLRs. Furthermore, TIR domain-containing adaptors, such as MyD88, TRIF, TIRAP and TRAM, play essential roles in TLR signaling. MyD88 is essential for inflammatory cytokine production via all TLRs, whereas TRIF is involved in TLR3- and TLR4-mediated MyD88-independent induction of IFN-beta. Thus, innate immunity represents a skillful system that senses microbial invasion and initiates appropriate immune responses. PMID- 15826380 TI - Purification and characterization of deacylated and/or palmitoylated lipid A species unique to Salmonella enterica serovar Typhimurium. AB - The Salmonella enterica serovar Typhimurium virulence gene products PhoP/PhoQ sense host microenvironments to regulate the expression of a lipid A 3-O deacylase, PagL, and a lipid A palmitoyltransferase, PagP. Therefore, deacylation and/or palmitoylation of lipid A could occur in Salmonellae adapted to host environments. The PhoP/PhoQ-regulated modification of lipid A alters host recognition and signaling, and may play an important role in host defense against Salmonellae infection. Here we report the purification and characterization of modified lipid A species. Deacylated lipid A, deacylated and palmitoylated lipid A, and palmitoylated lipid A species were generated in Escherichia coli cells heterologously expressing salmonellae PagL and/or PagP, and then purified by sequential thin-layer chromatography. The purified lipid A species showed m/z values that correspond to single lipid A species on mass spectrometry analysis. The modified lipid A species showed reduced ability to induce cellular signaling through Toll-like receptor 4, suggesting a specific function of the lipid A modifications in the pathogenesis of salmonellae infection. PMID- 15826381 TI - Otto H. E. Westphal (1913-2004). PMID- 15826383 TI - The heavy burden of a silent scourge: domestic violence. PMID- 15826384 TI - [Validation of a short questionnaire to use in clinical consultations to detect gender violence]. AB - OBJECTIVE: To validate a Spanish-language questionnaire to be used in primary care consultations to detect cases of violence against women. METHODS: The validation study was carried out between October 2002 and October 2003 in the Program of General Internal Medicine of the Clinical Hospital of the University of Buenos Aires, in the city of Buenos Aires, Argentina. The original version of this Spanish-language questionnaire had been developed in sexual and reproductive health clinics of five countries of Latin America and the Caribbean. The questionnaire covered the most frequent forms of gender-based violence (emotional, physical, sexual, and sexual abuse during childhood); explored how and when the violence occurred and who perpetrated it; and investigated the patient's perception of her current risk. In the research in Buenos Aires the format and theoretical content of the survey were evaluated, and linguistic and psychometric validations were also carried out. The reliability of the tool was assessed in terms of its reproducibility, stability, and internal consistency. RESULTS: The study found that the women surveyed in Buenos Aires had a good understanding of the questions, the examples given with the questions, and the response options. The women did not perceive the survey as being intimidating or as an intrusion into their private lives. The women generally regarded the use of the survey in primary care settings as a key opportunity for them to express their suffering and to receive assistance. The level of agreement for the answers to the questions in all five areas (psychological violence, physical violence, sexual violence, sexual abuse during childhood, and perception of current risk) was very high, in terms of both reproducibility (kappa values of 0.63 to 1.00) and stability (kappa values of 0.62 to 1.00). The alpha correlation coefficient for internal consistency was 0.755 for psychological violence and physical violence, 0.498 for psychological violence and sexual violence, 0.596 for physical violence and sexual violence, and 0.715 for the different manifestations of violence (psychological, physical, sexual, and abuse during childhood). CONCLUSION: This research demonstrated the soundness of the brief questionnaire, in terms of its format, content, comprehensiveness, and reliability. This instrument can be used in clinical consultations to detect situations of psychological violence, physical violence, sexual violence, and sexual abuse during childhood. PMID- 15826385 TI - [Risk factors for anemia among 6- to 12-month-old children in Brazil]. AB - OBJECTIVE: To estimate the prevalence of anemia and to determine associated risk factors among infants receiving routine health care in public clinics in Brazil. METHOD: This cross-sectional study included 2,715 infants between 6 and 12 months old in 12 cities, in all five of the geographic regions of Brazil. Information regarding the child and its feeding habits was obtained from the mother or other caregiver, using a questionnaire. Nutritional status was determined based on height and weight measurements. The hemoglobin concentration was measured using the HemoCue portable hemoglobinometer. Anemia was defined as hemoglobin < 11 g/dL. The infants' eating habits were assessed based on what they were eating around the time of the questionnaire interviews. The association between anemia and the different variables was evaluated through bivariate analysis, followed by multiple logistic regression using a hierarchical selection model. RESULTS: The prevalence of anemia for the entire group was 65.4%. Multiple regression analysis identified the following risk factors for anemia: living in the Southeastern Region of Brazil (odds ratio (OR) = 1.57, 95% confidence interval (95% CI) = 1.25 1.99), maternal age < 20 years (OR = 1.58, 95% CI = 1.21-2.07), birthweight < 2,500 g (OR = 2.09, 95% CI = 1.48-2.95), not being breast-fed (OR = 1.28, 95% CI = 1.02-1.61), receiving both breast milk and other foods (OR = 1.40, 95% CI = 1.10-1.78), and male gender (OR = 1.24, 95% CI = 1.06-1.46). CONCLUSIONS: The high proportion of anemic children indicates the need to emphasize, in prenatal and infant health programs, intervention measures for anemia control. Our results could guide these measures, focusing on the groups at greatest risk, such as low birthweight babies and the children of adolescent mothers. PMID- 15826386 TI - Changes in the AIDS epidemiologic situation in Puerto Rico following health care reform and the introduction of HAART. AB - OBJECTIVES: To compare the occurrence of AIDS as well as the sociodemographic and clinical profiles of AIDS patients in Puerto Rico before and after the introduction of highly active antiretroviral therapy (HAART) and the privatization of the island's public health care system. METHODS: We compared the incident AIDS cases for two three-year periods, 1992-1994 and 1998-2000, in four populations: (1) entire United States, (2) Puerto Rico, (3) Bayamon Health Region (located in north-central Puerto Rico, it includes 11 of the island's 78 municipalities), and (4) an HIV cohort enrolled at the Universidad Central del Caribe (UCC) School of Medicine. The UCC is located in Bayamon, Puerto Rico, within an academic medical complex that houses the teaching hospital (Ramon Ruiz Arnau University Hospital), the ambulatory health care facilities (Immunology Clinics) for patients with HIV, and administrative buildings. This represents the major government-sponsored health care infrastructure within the Bayamon Health Region. RESULTS: Incident AIDS declined substantially between the two periods in each of the four populations studied. The 48.1% decline in Puerto Rico exceeded the 40.9% decline in the United States. The decline in Puerto Rico likely resulted from increased availability and implementation of HAART and the delivery of health care to HIV/AIDS patients in an integrated fashion within each regional ambulatory clinic. In spite of this improvement, the absolute number of patients with AIDS on the island remains high. Substantial resources for treatment and prevention are required. The proportion of new AIDS cases was lower among women, persons 40 years of age or older, the less educated, and those living alone. Injection drug use remains the predominant mode of transmission in Puerto Rico. CONCLUSIONS: Further gains in Puerto Rico's fight against AIDS will depend on the island's ability to reduce the transmission that occurs through injection drug use; the use of HAART on a larger number of vulnerable patients, particularly intravenous drug users; educational interventions to improve medication compliance in certain risk groups; and specific measures aimed at decreasing the rate of injection drug use. PMID- 15826387 TI - [Atraumatic restorative treatment to control dental caries: history, characteristics, and contributions of the technique]. AB - This paper presents relevant scientific information on the history, characteristics, and contributions of atraumatic restorative treatment (ART) for use in preventing and controlling dental caries. Within the area of oral public health, ART has been for years an economical, effective method for preventing and controlling caries in vulnerable populations. Among other things, ART reduces the stress and anxiety in patients that conventional restoration methods produce. This technique promises major benefits for Latin America. However, given its limitations with dental cavities on two or more surfaces, it is recommended that more research on this approach be encouraged, with the aim of improving the technique's effectiveness based on its characteristics, indications, and technical merits. PMID- 15826388 TI - A paradigm for developing a comprehensive treatment protocol for survivors of domestic violence. PMID- 15826390 TI - Maternal death due to domestic violence: an unrecognized critical component of maternal mortality. PMID- 15826391 TI - Enhancing regional capacity in chronic disease surveillance in the Americas. PMID- 15826392 TI - [The most recent advances in developing vaccines against malaria]. PMID- 15826393 TI - [The prevalence of Chagas' disease in puerperal women and congenital transmission in an endemic area of Peru]. AB - OBJECTIVES: To determine the prevalence of antibodies against Trypanosoma cruzi in puerperal women and to assess possible congenital transmission of Chagas' disease in the department of Arequipa, Peru, where the disease is endemic. METHODS: Women who had given birth between December 2001 and July 2002 in three hospitals (two urban and one rural) and four health centers (three rural and one urban) of the department of Arequipa, Peru, were studied. The serological study included screening all the puerperal women in order to detect antibodies against T. cruzi through indirect immunofluorescence (IIF), with confirmatory testing done with enzyme-linked immunosorbent assay (ELISA) testing and the titration of immunoglobulin G (IgG) antibodies by IIF. IIF tests to screen for immunoglobulin M (IgM) antibodies were done with the seropositive women and their newborns, and infection was evaluated through xenodiagnosis (evaluated at 30 and 60 days) and the direct micromethod of Freilij et al. The results were analyzed in terms of the presence of the vector and of cases of Chagas' disease in the places where the puerperal women had been born and where they were living. Two neonatologists clinically evaluated the newborns in order to detect abnormalities and signs of congenital Chagas' disease. RESULTS: The overall prevalence of Chagas' disease in the 3 000 puerperal women studied was 0.73%. Prevalence was highest in two health centers located in rural areas (2.2% in El Pedregal and 4.1% in La Joya) (P=0.018). The disease was associated with previous direct contact with the vector (P<0.05) and with having been born in an area considered endemic (P<0.01). Four (20%) of the 20 seropositive puerperal women were also positive by xenodiagnosis. However, none of the women was aware of her infectious carrier state, and none showed the characteristic symptoms or signs of acute or chronic Chagas' disease. IgM antibodies were not detected in any of the puerperal women. One neonate (whose mother did not have evidence of parasitemia) presented an IgM titer of 1/8, but in later controls neither IgM nor IgG antibodies were detected. Parasites were not detected in the blood of the neonates by either of the two testing methods used. Of the 20 neonates evaluated, one presented microcephaly and hepatosplenomegaly; although the child had specific IgG antibodies against T. cruzi at birth, the antibodies were not present at the age of two months. The growth and development of the other 19 newborns were normal. CONCLUSIONS: The prevalence of Chagas' disease in puerperal women of the department of Arequipa, Peru, is low. No cases of intrauterine congenital transmission were found. We recommend carrying out studies on prenatal detection that evaluate more mothers and in which women who give birth at home also participate. PMID- 15826394 TI - HIV prevention programs of nongovernmental organizations in Latin America and the Caribbean: the Global AIDS Intervention Network project. AB - OBJECTIVE: The objective of this paper is to describe HIV prevention programs conducted by nongovernmental organizations (NGO) that are meeting this challenge. METHODS: One NGO undertaking HIV prevention programs was evaluated in each of the 23 countries participating in the Global AIDS Intervention Network (GAIN) Project throughout Latin America and the Caribbean. A two-stage selection process was used: (1) a search in databases and other information sources; (2) identification of NGOs that were best established and most actively engaged in HIV prevention activity. Executive directors were questioned about staffing, budget issues, populations served and barriers faced by these entities. RESULTS: The 23 NGOs conducted 58 direct-service programs and had been conducting HIV prevention activities for a mean of 8 years (SD=4.45; range 1-18 years). Average annual program budget was US $205,393 (range: US $10,000 to US $1,440,000). The NGOs reported a mean of 4.5 full-time employees (range 0-15, SD=4.7). Many relied on volunteers (median=10, mean=51, range 0-700, SD=150) to conduct HIV prevention activities. The NGOs provided prevention services for the general community (82.6%), children and adolescents (34.8%) and men who have sex with men (30.4%). Activities conducted by NGOs included train-the-trainer activities (43.5%) and face-to-face prevention activities (34.8%). Obstacles cited included lack of funding (60.9%) and HIV-related stigma and discrimination (56.5%). CONCLUSION: The strategies used by NGOs to overcome barriers to prevention are a testament to their ingenuity and commitment, and serve as examples for NGOs in other world regions. PMID- 15826395 TI - [Smoking in households in Brazil with children younger than 5 years of age]. AB - OBJECTIVES: To estimate the prevalence of exposure to smoking in households with children younger than 5 years of age in the city of Cuiaba, Mato Grosso, Brazil, and to identify the main determinants of that exposure. METHODS: This cross sectional, population-based study included 2 037 families from the city of Cuiaba. The parents of the children or other caregivers answered a questionnaire concerning the smoking habits of people living in the household, sociodemographic characteristics of the household, and the household's living conditions. RESULTS: The overall prevalence of smoking in the households was 37.7%. In terms of income, in the households with a monthly per capita income of 2 minimum wages or more, the prevalence of smoking was 24.1%; it was 31.3% in the households with a monthly per capita income between 1 and 1.9 minimum wages; and it was 46.0% in households with a monthly per capita income below 1 minimum wage. With respect to socioeconomic level (the family's material goods and purchasing power and the educational level of the head of the household, divided into five classes, from A (highest) to E (lowest)), the higher the socioeconomic level, the lower was the prevalence of smoking in the household: 26.9% for class A, 26.4% for class B, 34.9% for class C, 45.1% for class D, and 47.2% for class E. There was a higher prevalence of smoking when the father did not live in the home (47.5% versus 35.5%), when the head of the family was someone other than the father or the mother (53.8% versus 33.9% and 38.8%, respectively), when the father was younger than 20 years (52.3% versus 31.2% for fathers 40 or older), and when the mother was younger than 20 years (46.1% versus 22.5% for mothers 40 or older). In terms of occupation, the lowest prevalence of household smoking was found with health professionals and teachers, for both women (18.3%) and men (14.7%). After multivariate logistic regression analysis, the following variables remained associated with household smoking: absence of the father from the household, maternal and paternal schooling, maternal and paternal occupation, child's age, and household monthly per capita income. CONCLUSION: The risk of exposure to household smoking (and thus to passive smoking) in this sample of children younger than 5 years of age was higher in households living in poorer socioeconomic conditions. PMID- 15826396 TI - Frequency and characteristics of selected enteropathogens in fecal and rectal specimens from childhood diarrhea in Trinidad, 1998-2000. AB - OBJECTIVE: To determine the prevalence and characteristics of selected enteric pathogens in diarrheic children in six counties of the island of Trinidad. METHODS: This cross-sectional study was conducted from April 1998 through March 2000. Fecal or rectal swab specimens from children (<12 years) were collected and then processed, using standard methods, to detect Salmonella spp., Shigella spp., enteropathogenic Escherichia coli (EPEC), Campylobacter spp., Yersinia spp., Cryptosporidium parvum, and parasite ova. The antibiograms of the enteropathogens were determined using the disk diffusion method. RESULTS: A total of 236 samples were processed; 86 samples originated directly from 17 heath centers in two counties (St. George East and St. George West), while 150 samples were obtained from the Trinidad Public Health Laboratory, having been submitted by private practitioners and personnel from six counties in Trinidad. Of the 236 samples, 33 (14.0%) were positive for Shigella, 4 (1.7%) for Salmonella, and 1 (0.4%) for EPEC. Two of the samples (0.8%) were positive for Campylobacter jejuni, while 1 sample (0.4%) was positive for hookworm ova. All the samples were negative for Cryptosporidium parvum and Yersinia spp. With the 86 samples collected directly from the health centers, in St. George East County the frequency of Shigella was 20.0% (12 of 60), compared with 26.9% (7 of 26) for samples from St. George West County, but the difference was not statistically significant (P>0.05 with the chi square test). For the 150 samples from the six counties that had been submitted directly to the Trinidad Public Health Laboratory, 14 of them (9.3%) were positive for Shigella, a figure statistically significantly lower than that found with the samples sampled directly from the health centers (P<0.05 with the chi square test). Sh. sonnei was the predominant serotype detected, accounting for 28 of the 33 Shigella isolates (84.8%) recovered from the 236 samples. Overall, the frequency of detection of enteropathogens had no seasonal pattern nor relationship to the county of origin. Of the 37 isolates of Salmonella and Shigella tested for antimicrobial sensitivity, all of them were sensitive to ciprofloxacin, gentamicin, and cefotaxime. In terms of resistance, 3 of the 37 isolates (8.1%) exhibited resistance to ampicillin, 1 (2.7%) to chloramphenicol, and 1 (2.7%) to sulfamethoxazole/ trimethoprim. CONCLUSION: Of the enteropathogens for which assays were done, Shigella sonnei was the most prevalent, and it has the highest probability of being an important etiological agent of childhood diarrhea in Trinidad. PMID- 15826397 TI - [The economic costs of pneumonia in children under 2 years of age in Colombia]. AB - OBJECTIVE: To analyze the costs of pneumonias presumed to be of viral or bacterial origin, as diagnosed by chest X ray, in four reference center hospitals in three cities in Colombia. METHODS: A total of 128 cases of pneumonia (64 bacterial cases and 64 viral cases) that had consecutively entered the hospitals in the study between July 2001 and January 2003 were investigated. The diagnosis of pneumonia was based on chest X rays. The study population was composed of children under 2 years of age who required hospitalization. In order to estimate the costs for bacterial pneumonias and viral pneumonias, the cost of each activity was determined for each case, and then average costs were calculated. RESULTS: The average cost of the presumably bacterial pneumonia cases was US $611.50 (95% confidence interval (95% CI), US $532.20-690.80); that of the presumably viral cases was US $472.20 (95% CI, US $331.80-612.60). The observed differences were due to direct expenses, especially drugs (antibiotics), special services, and diagnostic tests. In the two groups the families were similar in their incomes and the indirect costs that they had to bear, so the indirect costs were not considered relevant in terms of distinguishing between the costs caused by the two forms of pneumonia. CONCLUSIONS: The study found differences in the direct costs of care between the presumably bacterial cases and the presumably viral ones. The study results also gave an approximation of the indirect costs to the patients' families caused by the pneumonias. The differences found in the direct costs also indicate that X-ray diagnosis is useful for differentiating between viral and bacterial pneumonia. Few studies in Latin America have assessed the economic costs of pneumonia in children, so this study can serve as a reference for future research on the impact of interventions against pneumonia. PMID- 15826398 TI - [Immunization coverage survey: empirical assessment of the cluster sampling method proposed by the World Health Organization]. AB - OBJECTIVE: To assess sample representativeness and the precision of estimates of immunization coverage obtained with the 30 by 7 cluster sampling method proposed by the World Health Organization, by applying the method to determine immunization coverage in two municipalities (Diadema and Sao Caetano do Sul) in the state of Sao Paulo, Brazil, in 2000. METHOD: The representativeness of the samples was determined by comparing the census sectors picked by lot for the surveyed sectors and for the nonsurveyed sectors in both municipalities, in terms of socioeconomic and demographic characteristics (age distribution of the population, schooling, proportion of households headed by a women, monthly income of household head, and sanitary conditions of the home (piped-in water, connected to the sewer system)). The precision of the coverage estimates for the vaccines in the basic immunization schedule-BCG; diphtheria, pertussis, and tetanus (DPT); poliomyelitis; hepatitis B; measles; and measles, mumps, and rubella (MMR)-was determined by calculating the design effect and the width of the confidence intervals. Precision was considered to be satisfactory if the design effect was below 2.0 and the confidence interval width was below 10%. RESULTS: In both municipalities the comparison between the surveyed and nonsurveyed sectors showed a similar distribution in terms of socioeconomic and demographic variables. Concerning the precision of the estimates, the design effect was below 2.0 for all the vaccines, both in Sao Caetano do Sul and Diadema. In Diadema, the confidence interval width was below 10% for all the vaccines, except for MMR (10.1%). In Sao Caetano do Sul, only 89% of the expected sample were included, so the width of the confidence interval was slightly above 10% for the poliomyelitis vaccine (10.3%), the hepatitis B vaccine (11.8%), the mumps vaccine (10.4%), the MMR (12.9%), and the complete schedule (11.2%). CONCLUSION: The cluster sampling method proposed by the World Health Organization produces representative data as long as the methodological procedures for selecting the sample are rigorously followed in the field. PMID- 15826399 TI - Asthma control in Latin America: the Asthma Insights and Reality in Latin America (AIRLA) survey. AB - OBJECTIVES: The aims of this survey were (1) to assess the quality of asthma treatment and control in Latin America, (2) to determine how closely asthma management guidelines are being followed, and (3) to assess perception, knowledge and attitudes related to asthma in Latin America. METHODS: We surveyed a household sample of 2,184 adults or parents of children with asthma in 2003 in 11 countries in Latin America. Respondents were asked about healthcare utilization, symptom severity, activity limitations and medication use. RESULTS: Daytime asthma symptoms were reported by 56% of the respondents, and 51% reported being awakened by their asthma at night. More than half of those surveyed had been hospitalized, attended a hospital emergency service or made unscheduled emergency visits to other healthcare facilities for asthma during the previous year. Patient perception of asthma control did not match symptom severity, even in patients with severe persistent asthma, 44.7% of whom regarded their disease as being well or completely controlled. Only 2.4% (2.3% adults and 2.6% children) met all criteria for asthma control. Although 37% reported treatment with prescription medications, only 6% were using inhaled corticosteroids. Most adults (79%) and children (68%) in this survey reported that asthma symptoms limited their activities. Absence from school and work was reported by 58% of the children and 31% of adults, respectively. CONCLUSIONS: Asthma control in Latin America falls short of goals in international guidelines, and in many aspects asthma care and control in Latin America suffer from the same shortcomings as in other areas of the world. PMID- 15826401 TI - [Neoliberal health sector reforms in Latin America: unprepared managers and unhappy workers]. AB - This work analyzes the neoliberal health sector reforms that have taken place in Latin America, the preparation of health care workers for the reforms, the reforms' impacts on the workers, and the consequences that the reforms have had on efficiency and quality in the health sector. The piece also looks at the process of formulating and implementing the reforms. The piece utilizes secondary sources and in-depth interviews with health sector managers in Bolivia, Colombia, Costa Rica, the Dominican Republic, Ecuador, El Salvador, and Mexico. Neoliberal reforms have not solved the human resources problems that health sector evaluations and academic studies had identified as the leading causes of health system inefficiency and low-quality services that existed before the reforms. The reforms worsened the situation by putting new pressures on health personnel, in terms of both the lack of necessary training to face the challenges that came with the reforms and efforts to take away from workers the rights and benefits that they had gained during years of struggles by unions, and to replace them with temporary contracts, reduced job security, and lower benefits. The secrecy with which the reforms were developed and applied made workers even more unified. In response, unions opposed the reforms, and in some countries they were able to delay the reforms. The neoliberal reforms have not improved the efficiency or quality of health systems in Latin America despite the resources that have been invested. Nor have the neoliberal reforms supported specific changes that have been applied in the public sector and that have demonstrated their ability to solve important health problems. These specific changes have produced better results than the neoliberal reforms, and at a lower cost. PMID- 15826402 TI - [Neoliberal reforms in health services in Latin America: a critical view from two case studies]. AB - Neoliberal reforms have promoted privatization and decentralization as strategies to improve equity, efficiency, and the quality of health services. In this piece the impact of these reforms in Latin America is critically analyzed, and the impacts of privatization in Colombia and of decentralization in Mexico are detailed. These two cases show that after 10 years of privatization in Colombia and 20 years of decentralization in Mexico the reforms have had the opposite of the desired effect: They have not improved equity, have increased health expenditures, have not increased efficiency, and have not shown a positive impact on quality. Public health programs in Colombia have deteriorated, while decentralization in Mexico has had a very high cost, without achieving the proposed objectives. It is officially accepted that decentralization in Mexico has increased inequity, and that new reforms implemented in 2003 promote vertical programs. Health systems based on regulated competition are not the most suitable ones for Latin America. Latin American countries should improve their health systems in line with the principles stated in the Declaration of Alma Ata and according to their own national experiences. PMID- 15826403 TI - A Conversational Model of art therapy. AB - This paper illustrates a 'Conversational Model' of art therapy. The Conversational Model was jointly created by Robert Hobson and Russell Meares. It is a developmental theory unique in its clinical application. The focus of the paper is two sessions that altered the course of therapy. In these sessions, variations on Donald Winnicott's "squiggle-game" and Hobson's "party game" were used to engage an isolative, reluctant incarcerated patient. The interventions illustrate the basic tenets of the Conversational Model. The theoretical process- from disruption to repair--is visually recorded in the artwork. The central argument of the paper is that interactive art therapy interventions can be effective, when used appropriately. By engaging the patient in a 'visual' conversation, he/she may develop an emotional vocabulary, a prerequisite for a psychotherapeutic conversation. The paper begins with a brief historical overview of the interface between art and psychoanalysis, the context out of which 'art therapy'--a distinct body of theory--evolved. Theory interweaves with clinical material in a narrative style. What I say and do in therapy is aimed at promoting understanding: a 'conversation', a meeting between two experiencing subjects (an I and a Thou), here and now, in such a way that learning can be effective in other relationships. If, as I believe, psychotherapy is a matter of promoting a personal dialogue, then we need to know how to receive, express, and share feeling: how to learn a language of the heart in its 'minute particulars'. PMID- 15826404 TI - Vocal manifestations of internal multiplicity: Mary's voices. AB - According to the assimilation model, personality may be considered as a community of internal voices representing traces of previous experiences. This study addressed the hypothesis that a person's internal voices sound different from each other. First, following an intensive qualitative procedure, four clinically sophisticated raters identified six internal voices in a 45-minute interview with Mary, a graduate student in psychology. They characterized the voices by (a) distinctive names, (b) descriptions of vocal and personality characteristics, and (c) sets of excerpted passages. Next, using the voice names and descriptions, additional raters independently sorted the passages into groups. Sortings based on transcript plus audio agreed with the sophisticated raters on 68.9% of the passages, as compared with 37.2% for sortings based on transcript only, while chance agreement expected for both was 16.7%. These findings support a literal view of internal multiplicity, consistent with a modular organization of experiential information within personality. PMID- 15826405 TI - The effect of mood on responses to the Young Schema Questionnaire: short form. AB - The schema concept has had an important role in both early and more recent accounts of psychopathology. Schemas are underlying cognitive structures that are thought to create vulnerability to disorders, because they act as templates for the perception, encoding, storage, and retrieval of information. Recent approaches to the understanding and treatment of personality disorders give schemas particular prominence, and the concept of early maladaptive schemas forms the cornerstone of Young's (1999) schema-focused cognitive therapy. The aim of this paper is to examine the effect of mood on responses to the Young Schema Questionnaire--short form (YSQ-S; Young, 1998). A sample of 50 non-clinical participants completed the YSQ-S on three different occasions: in neutral mood, and following happy and depressed mood inductions. The results of 30 participants with full data sets showed that emotional deprivation and defectiveness scores increased after the depressed mood induction, whereas entitlement scores increased after the happy mood induction. The results are discussed in relation to cognitive theories of vulnerability to psychopathology, and future directions for research are suggested. PMID- 15826406 TI - Ex-clients' evaluation of bereavement counselling in a voluntary sector agency. AB - This study aims to carry out an evaluation of bereavement counselling provided by a voluntary charity from the perspective of clients who had availed themselves of the service. A questionnaire survey of ex-clients of one branch of a major voluntary sector bereavement care organization was carried out. Based on a sample of 89 returned questionnaires (representing a response rate of 45%), findings indicated that paraprofessional voluntary counsellors can provide a counselling service to bereaved people that satisfies the majority of clients who report positive experiences in counselling and post-counselling outcomes, and generally rate their counselling as helpful. Nonetheless, there was a minority of ex clients who did not regard their status, 6 weeks after counselling, as being more positive. Further, the reputation of the voluntary agency within the community and among potential referrers and ex-clients appears strong, offering additional positive evidence of the status of their bereavement counselling service in the locality. PMID- 15826407 TI - Protecting the sacred and expressions of rituality: examining the relationship between extrinsic dimensions of religiosity and unhealthy guilt. AB - The aim of the present study was to examine the relationship between extrinsic religiosity and unhealthy guilt by testing two specific hypotheses. Using multidimensional measures of religious orientation and guilt, among 344 (169 male and 175 female) adults, and consistent with predictions; extrinsic-personal religiosity was found to be related to trait and state guilt, and extrinsic social religiosity was found to be related to moral standards guilt. The present findings suggest evidence for a present conceptualization of the relationship between extrinsic religiosity and unhealthy guilt as the result of internal and external conflicts. The findings suggest that, when looking at maladaptive guilt among religious clients in clinical, therapeutic or counselling settings, it may be crucial to distinguish between dimensions of extrinsic religiosity and unhealthy guilt. PMID- 15826408 TI - The relationship between social deprivation and unilateral termination (attrition) from psychotherapy at various stages of the health care pathway. AB - The relationship between social deprivation and attrition from psychotherapy was examined at various stages of the health care pathway. Data providing information on service users' discharge status gave a measure of attrition at different stages along this pathway. On the basis of their postcode, service users were allocated a Townsend deprivation score, which is a measure of social deprivation. Of the sample, 60% terminated therapy prior to agreed discharge at various pathway stages. Service users who failed to attend their first appointment had significantly higher social deprivation scores than those who completed therapy. Early terminators, who stopped attending before their fifth session, had a significantly higher level of social deprivation than the late terminators, and those who completed therapy. The late terminators did not significantly differ from completers on social deprivation. These results support earlier findings showing that socio-economic status influences attrition from therapy. However, socioeconomic status only affects two stages of the pathway - attendance at the first appointment, and the early stage of attending therapy. It does not affect earlier or later stages of the pathway. Applying the health action process approach (HAPA) model to the results, the action/maintenance stage can be represented by attendance for therapy. Socio-economic status influences this stage of the model, because social support and resources are important determinants for compliance. Earlier stages, characterized by HAPA as a decisional/motivation stage thought to be influenced by beliefs, was not affected by social deprivation. It is concluded that attrition from therapy should be studied separately for each pathway stage. Earlier conflicting findings on causes of attrition may have resulted from studying different combinations of pathway stages. PMID- 15826409 TI - Alexithymia: its prevalence and correlates in a British undergraduate sample. AB - INTRODUCTION: Alexithymia is characterized by a difficulty identifying and describing emotional states, as well as an externally oriented thinking style. This study investigated the prevalence of alexithymia in a British undergraduate sample and assesses its relationship to both parental bonding and dissociation. METHOD: The Toronto alexithymia scale (TAS-20), the Parental Bonding Instrument (PBI), and the Dissociative Experience Scale (DES) were administered to a sample of 181 male and 190 female undergraduate students from both arts and science subjects. RESULTS: Rates of alexithymia were comparable with those observed in some other countries. Contrary to predictions, females were found to have higher rates than males, and the highest presence of alexithymia was in female science students. As in previous studies, alexithymia was linked to both dissociation and perceptions of a lack of maternal care, though the degree of association to the latter was small. Dissociative experiences were predicted by both maternal overprotection and difficulties identifying feelings. DISCUSSION: Some qualified support was found for the relevance of early maternal bonding to later difficulties processing emotions. The presence of greater alexithymia in females, and female science students in particular, was discussed in reference to similar observations elsewhere. There was also an understandable relationship between 'difficulty identifying feelings' (TAS) and both depersonalization/derealization and absorption (DES). PMID- 15826410 TI - Thirty-six month outcome data from a trial of counselling with chronically depressed patients in a general practice setting. AB - Counsellors have been employed in general practice with little evidence of effectiveness in this setting. This randomized controlled trial examined long term effectiveness of short-term counselling in general practice for patients with chronic depression, either alone or combined with anxiety. Participants were 181 patients recruited from nine general practitioners' (GP) practices in Derbyshire by screening consecutive attenders using the Beck Depression Inventory. Both the experimental and control group received routine GP treatment but the experimental group were also referred to the practice counsellor. Depression and social adjustment were measured at baseline, 6, 12, and 36 months. There was an overall significant improvement in the actual scores over time, but there were no significant differences between the two groups on any of the measures at either 6, 12, or 36 months. Although fewer experimental group patients were still 'cases' on the BDI than controls at 6 and 12 months, this difference disappeared at 36 months. There was no evidence to demonstrate a long term effect of improved outcomes in those referred to counselling. PMID- 15826412 TI - The nutrition of active muscle flaps. AB - In the last 25 years, techniques utilising muscle flaps to perform mechanical work have become part of the reconstructive surgeon's repertoire. In this paper, these techniques and the particular physiological challenges faced by active flaps are discussed and a sub-classification of muscle flaps into passive and active is proposed. A series of experiments are described that were designed to compare local pO(2) and gaseous perfusion, at multiple sites in pedicled tibialis anterior muscle flaps and controls, using specially developed dual amperometric microelectrodes, in an in vivo rabbit model. No significant difference in perfusion was found comparing flaps and controls until the flap's vascular pedicles were divided and ligated, and then flap perfusion fell to zero. Flap and control oxygenation varied significantly only during mobilisation and again, after pedicle division and ligation (F[13,55] = 12.3; P < 0.001)]. This absence of functional ischaemia as a consequence of mobilisation in the model used suggests the practice of delaying electrical conditioning of muscle flaps in dynamic cardiomyoplasty and neoanal sphincteroplasty might be unnecessary if these data are reproduced in human latissimus dorsi muscle. The relevance of these findings to free flap oxygen consumption and to the delay phenomenon is discussed. PMID- 15826413 TI - Disease-centred advice for patients with superficial transitional cell carcinoma of the bladder. AB - INTRODUCTION: Urologists tend to regard superficial tumours (e.g. pTa or pT1 transitional cell carcinoma of the bladder) as being of low pathogenicity. There is a clearly established link between cigarette smoking and bladder cancer, with incidence, recurrence and mortality being positively associated with duration of smoking and number of cigarettes smoked. PATIENTS AND METHODS: A questionnaire based audit was undertaken to determine the amount of information being provided by urologists for patients who had been diagnosed with pTa or pT1 transitional cell carcinoma of the bladder about both their disease, its aetiology and appropriate advice regarding life-style change. RESULTS: A total of 78 patients adequately completed the questionnaire. Of these, 55 (71%) had been smokers at some time, and 24 (31%) continued to smoke at the time of follow-up. Only 26 of these 55 (47%) were aware of their underlying diagnosis. This level of knowledge was similar in non-smokers, of whom only 12 (52%) were aware of their disease. Of the ever-smokers, only 12 (22%) were aware that smoking was a risk factor for the development of bladder cancer, and 7 (13%) were aware that continued smoking could worsen the prognosis. Only 18 (33%) of the 55 smoking patients had been told to stop smoking, for any reason, by their general medical practitioner, and only 4 (7%) had been told to stop by a urologist. CONCLUSION: In the urology department in which the audit was performed, patients with bladder cancer were not being provided with adequate information about their disease. PMID- 15826414 TI - Screening for prostate cancer: the case for. PMID- 15826415 TI - Radioguided occult lesion localisation (ROLL) is available in the UK for impalpable breast lesions. AB - INTRODUCTION: To test the feasibility and reliability of ROLL in a district general hospital (DGH) dealing with screening detected breast lesions. PATIENTS AND METHODS: [(99m) Tc]-labelled colloidal human serum albumin was injected in the core of the breast lesion under ultrasound or stereotactic guidance 2-4 h prior to surgery. At operation, the radioactivity is localised using a gamma probe. This allows optimal placement of the skin incision and subsequent WLE of the abnormal area. RESULTS: ROLL was utilised on 36 patients (median age, 61 years; range, 43-75 years); of these, 33 B5 lesions had a therapeutic one-step procedure (lumpectomy and axillary dissection) and 3 B4 patients had the lesion excised for diagnostic purposes. Localisation lasted a median of 8 min (range, 5 15 min), ROLL-guided wide tumour excision lasted 20 min (range, 15-30 min), and median postoperative hospital stay was 2 days (range, 1-3 days). Median cancer diameter was 12 mm (range, 6-40 mm). Margins were clear in 29 patients, while 7 patients with DCIS had involved margins. Median minimal clearance was 5 mm (range, < 1-10 mm). Patients had either excellent (24/36) or good (12/36) cosmetic results. CONCLUSIONS: ROLL successfully localised all lesions; this technique can be implemented in any DGH with a Nuclear Medicine Department. The learning curve is short, cost effectiveness is proven, and cosmetic results are highly rewarding. ROLL could rapidly become the standard localisation technique in the UK. PMID- 15826416 TI - Day-case breast cancer axillary surgery. AB - INTRODUCTION: The standard locoregional management of breast cancer is excision of the primary tumour and axillary staging with suction drainage of the axilla. The objective of this study was to determine the safety, tolerability and efficacy of day-case surgery without suction drainage. PATIENTS AND METHODS: A review of complete, prospectively collected data was performed on all breast cancer patients (screening and symptomatic) planned to undergo day-case axillary surgery at a University Teaching Hospital between 2000 and 2002. Postoperative complications were recorded and the notes of patients not discharged on the day of their surgery were also examined to establish the reason for overnight stay. RESULTS: 165 patients underwent intended day-case axillary surgery (axillary dissection level 1/2; median age, 55 years; range, 39-76 years). Of these, 16 (9.7%) were admitted overnight usually due to over-running of theatre lists (n = 13; 81%). 29 patients (17.6%) underwent axillary dissection alone, the remainder had axillary surgery combined with wide local excision (median number of lymph nodes excised 11; range, 2-18). Complications included symptomatic seroma formation in 37 patients (22%) and wound infection in 16 patients (10%). CONCLUSIONS: Day-case axillary surgery can be performed safely with surgical morbidity comparing favourably to published work of 'traditional' axillary drainage following lymphadenectomy. PMID- 15826417 TI - The use of tranexamic acid to reduce blood loss during total hip arthroplasty: an observational study. AB - INTRODUCTION: To test the hypothesis that the fall in haemoglobin following total hip arthroplasty is reduced by tranexamic acid administration. PATIENTS AND METHODS: A cohort of 64 patients were studied, 32 received tranexamic acid 20 mg/kg on induction. Surgery was performed by the senior author in a standardised fashion. Haemoglobin levels were measured 2 weeks pre- and 3 days postoperatively. Any complications were noted. The study group was matched using the bone and joint research database for age, sex, procedure, disease and pre operative haemoglobin level. RESULTS: In the group receiving no tranexamic acid, the mean fall in haemoglobin was 3.8 g/dl (CI of mean 3.4-4.3) and in the group treated with tranexamic acid 2.8 g/dl (CI of mean 2.5-3.2) P < 0.05. Complications included one non-fatal pulmonary embolus in the tranexamic acid group. CONCLUSIONS: The administration of 20 mg/kg of tranexamic acid on induction of surgery is an effective method of reducing the haemoglobin fall following hip arthroplasty. PMID- 15826418 TI - An economic justification for autologous blood re-infusion in primary total knee replacement surgery. AB - INTRODUCTION: To justify economically the use of autologous blood re-infusion after total knee replacement surgery compared with vacuum drains. To determine if the patients using autologous re-infusion units have a reduced allogenic blood transfusion requirement and hospital stay. PATIENTS AND METHODS: Prospectively, 50 patients undergoing primary unilateral total knee replacements with autologous re-infusion units were studied. They were matched for age, sex, type of prosthesis and the month in which surgery took place to a second group undergoing the same surgery with vacuum drains. The results for the second group were obtained retrospectively from the notes. The outcome measures were the need for allogenic blood transfusion and length of postoperative hospital stay. RESULTS: The use of re-infusion units reduced the need for postoperative allogenic blood transfusion from 28% to 4% in total knee replacement surgery. The cost of using re-infusion units was the same as vacuum drains. Overall, the autologous re infusion patients were discharged 2 days earlier (99% confidence interval). CONCLUSIONS: Re-infusion units are no more expensive than vacuum drains. In addition, autologous blood has many clinical benefits compared to allogenic blood. Re-infusion may shorten the hospital stay for patients undergoing total knee replacement surgery. PMID- 15826419 TI - Satisfaction levels in orthopaedic out-patients. AB - INTRODUCTION: It is important that patients are satisfied with an out-patient consultation. This ensures compliance with treatment and attendance for follow up. The aim of this study was to identify factors regarding out-patient consultation associated with patient satisfaction. METHODS: A two-part questionnaire identifying expectations of, and subsequent satisfaction with, a new out-patient consultation was completed by 106 out-patients. RESULTS: There was no correlation between not seeing the clinician anticipated and reduced satisfaction (P = 0.17). Using more information sources was associated with less satisfaction (P = 0.02). Patients were less satisfied if their expectations of either treatment or outcome were changed. CONCLUSIONS: Meeting patients' expectations is an essential part of effective communication. The use of specialist physiotherapists and general practitioners with a special interest is an effective way of seeing more new patients. PMID- 15826420 TI - Shortcomings of the National Joint Registry: a survey of consultants' views. AB - INTRODUCTION: The National Joint Registry (NJR) for England and Wales was launched in April 2003. The UK Department of Health (DoH) awarded the contract to run the NJR to Atomic Energy Authority (AEA) Technology in September 2002. The aim was to etablish the views of a large group of orthopaedic consultants on the new NJR. METHODS: A questionnaire was sent by post to 405 orthopaedic consultants in the Midlands and South West. RESULTS: Overwhelming support was found for the idea of a national joint replacement register that is used for peer-run audit. However, there was wide-spread concern about the lack of orthopaedic representation on the steering committee. The majority of surgeons have concerns about the possible use of NJR data for the production of league tables. PMID- 15826422 TI - A method for determining the rate of major limb amputations in battle casualties: experiences of a British Field Hospital in Iraq, 2003. AB - INTRODUCTION: The majority of battle casualties undergoing surgery at 34 Field Hospital, the sole Coalition field hospital in Iraq during the conflict, sustained injuries to the extremities. To compare our experiences with those from previous conflicts, we report data on major limb amputations and propose a method for determining the rate of major limb amputation in a conflict setting. PATIENTS AND METHODS: A retrospective review of battle casualties admitted to the hospital was carried out based on casualty records and operating theatre logbooks. Data were collected for the period 26 March and 8 May 2003, focusing on casualties undergoing surgery for battle-injured extremities during the conflict. RESULTS: 68 (55%) casualties underwent surgery for battle injuries to extremities. Six upper and eight lower limb amputations (proximal to carpals and tarsals) were carried out from a total of 87 battle-injured limbs that had surgery, giving an overall amputation rate of 16% (14/87). CONCLUSIONS: In presenting our amputation rate of 16%, we highlight the lack of uniformity in describing 'amputation rates' between conflicts. A consistent method for quantifying amputations performed in a conflict setting could prove to be a useful tool. PMID- 15826423 TI - Primary total knee replacement: a comparison of a nationally agreed guide to best practice and current surgical technique as determined by the North West Regional Arthroplasty Register. AB - INTRODUCTION: In 1999, a statement of best practice in primary total hip replacement was approved by the Council of the British Orthopaedic Association (BOA) and by the British Association for Surgery of the Knee (BASK) to provide a basis for regional and national auditable standards: we have compared practice in the North West of England to this document to ascertain adherence to this guide to best practice. MATERIALS AND METHODS: A direct comparison of data held on the North West Hip Arthroplasty Register for 2001/2002 and BASK/BOA guidelines was performed. 86 surgeons from 26 hospitals were included in the study. RESULTS: A mean of 93.3% of operations were performed in the surgeon's usual theatre. All of these theatres had vertical laminar air flow systems. 42.2% of respondents routinely used exhaust suits and 68.1% of respondents routinely used impermeable disposable gowns. All surgeons use some form of anti-thromboembolic prophylaxis; 66.2% use a combination of both mechanical and chemical means. All surgeons used antibiotic prophylaxis. The most popular choice of antibiotic was a cephalosporin. 93.7% of surgeons routinely use antibiotic-loaded cement. The PFC and Kinemax prostheses were the most commonly used prostheses. Interestingly, 97.7% of all first-choice implants were cemented. Only 2 surgeons used uncemented total knee replacement. 69.8% of surgeons used a posterior cruciate retaining design. A midline longitudinal skin incision is used by 87.2% of surgeons, a medial longitudinal skin incision by 7.0% and a lateral longitudinal skin incision by 5.8% of surgeons. A medial parapatellar capsular incision is preferred by 91.9% with the remainder using mid vastus or trivector retaining capsulotomy. Closure of capsulotomies is performed in flexion by 65.1% and in extension by 34.9%. In patients with osteoarthritis, 38.4% routinely resurfaced the patella, 34.9% never resurfaced the patella and 26.7% selectively resurfaced. This was in direct contrast to practice for patients with rheumatoid arthritis in whom 66.3% routinely resurfaced the patella, 22.1% never resurfaced the patella and 11.6% selectively resurfaced. DISCUSSION AND CONCLUSIONS: This study has demonstrated variation of practice in hip arthroplasty across the North West region and significant divergence from the BASK/BOA statement of best practice. The introduction of a properly funded national arthroplasty register will surely help to clarify the effect of such diverse practice on patient outcome. PMID- 15826424 TI - Reducing methicillin-resistant Staphylococcus aureus (MRSA) patient exposure by infection control measures. AB - INTRODUCTION: To assess the effectiveness of infection control measures (pre admission screening and patient segregation) on reducing in-patient exposure to methicillin-resistant Staphylococcus aureus (MRSA). PATIENTS AND METHODS: A prospective case-control study in a district general hospital. All admissions to 3 wards over an 83-month period from September 1995 to July 2002 inclusive (a total of approximately 34,000 patients). Outcome measures were a statistical analysis of the difference in numbers of new cases of MRSA colonisation or infection between the 3 wards. RESULTS: There was a statistical significance in incidence of new MRSA cases between the ward with active infection control measures in place and the control wards. CONCLUSIONS: The described infection control measures reduced the exposure of patients to MRSA. This reduces the risk of MRSA infection, which is of importance in orthopaedics, and has further benefits that may also be applied in other surgical specialties. PMID- 15826425 TI - Intravenous fluid-associated morbidity in postoperative patients. AB - INTRODUCTION: There is marked variation in postoperative fluid prescribing which may contribute to postoperative morbidity. However, there are few data regarding the overall incidence of fluid associated morbidity in postoperative patients. PATIENTS AND METHODS: Data regarding fluid and electrolyte prescription, fluid balance and intravenous fluid associated morbidity were prospectively collected from 71 patients representing 173 patient days of intravenous fluid therapy. RESULTS: There was no correlation between fluid and electrolytes prescription and pre-operative weight, serum electrolyte levels or ongoing fluid losses. 17% of patients developed significant fluid associated morbidity. 7 patients developed a tachyarrhythmia, which was associated with the prescription of inadequate maintenance potassium. 5 patients developed fluid overload, associated with excessive fluid volume and sodium administration. CONCLUSIONS: Surgical house staff do not appear to use the available fluid balance information when prescribing. The introduction of fluid prescribing protocols may improve practice. This study provides an accurate measure of fluid-associated morbidity in order to measure the efficacy of such protocols. PMID- 15826426 TI - Vasectomy management in Morecambe Bay NHS Trust. AB - INTRODUCTION: Vasectomy is a common method of contraception in the UK. However, there is a wide variation in management protocols. The aim of the present study was to identify differences within the hospitals of Morecambe Bay NHS Trust and to recommend a uniform practice. PATIENTS AND METHODS: Retrospective case notes review of 395 vasectomy procedures performed within the Morecambe Bay NHS Trust in a 1-year period. RESULTS: Inconsistency was found with regards to the anaesthetic technique, the vas histology request and the timing of the semen analysis. The non-compliance rate for postvasectomy semen analysis was 33.4%. The complication and failure rates were 4.04% and 0.51%, respectively. Motile sperm (n = 4) was submitted at an average time of 8 weeks' postvasectomy. In half of those cases, vasectomy proved unsuccessful. Immotile sperm (n = 41) was submitted at an average time of 9.5 weeks and, in 80% of those men, semen cleared at an average time of 15.5 weeks' postvasectomy. An azoospermic (n = 285) sample was submitted at an average time of 10.5 weeks. Eleven of those men submitted a second sample with immotile sperm at an average time of 12 weeks' postvasectomy and that was eventually clear at 18 weeks in the majority of cases. CONCLUSIONS: A uniform vasectomy practice should include vasectomy under local anaesthesia if possible, no vas histology and a request for a single sample at 12 weeks. If this is clear, vasectomy should be considered successful. If any sperm are present, then a further sample should be requested at 16 weeks' postvasectomy. Immotile sperm at that time should not justify any further samples and a 'special clearance' should be issued to those men. PMID- 15826427 TI - Recent NICE guidance of interest to surgeons. PMID- 15826430 TI - Hepatitis C and general practice: the crucial role of primary care in stemming the epidemic. PMID- 15826431 TI - The prevention of suicide in later life: a task for GPs? PMID- 15826432 TI - Impact of a positive hepatitis C diagnosis on homeless injecting drug users: a qualitative study. AB - BACKGROUND: Increasing numbers of injecting drug users are presenting to primary care and a growing number of general practices are specifically providing care for homeless people. Injecting drug users are at the greatest risk of hepatitis C infection and homeless drug misusers, because of their drug-taking behaviour and patterns, have been identified as being at greater risk of harm of blood-borne diseases than the general population. However, little work has been conducted with injecting drug users or homeless people who have hepatitis C and little is known about how the virus may affect them. AIM: To explore the impact of a positive hepatitis C diagnosis on homeless injecting drug users. DESIGN OF STUDY: This study employed qualitative research. In-depth interviews allowed the exploration of the impact of a potentially life-threatening diagnosis within the context of a person's expressed hierarchy of needs. SETTING: A primary care centre for homeless people in the north of England. METHOD: In-depth interviews about the impact of a positive hepatitis C diagnosis on their lives were conducted with 17 homeless injecting drug users who had received a positive hepatitis C diagnosis. The interviews were audiotaped, transcribed, and analysed using the framework approach. RESULTS: Receiving a positive diagnosis for hepatitis C resulted in feelings of shock, devastation, disbelief, anger, and questioning. A positive diagnosis had lasting social, emotional, psychological, behavioural, and physical effects on homeless injecting drug users, even years after the initial diagnosis. Most responders were diagnosed by a doctor in primary care or by hospital staff; however, not all had sought testing and a number were tested while inpatients and were unaware that blood had been taken for hepatitis C virus serology. CONCLUSIONS: The implications for clinical policy and primary care practice are discussed, including the issues of patient choice, confidentiality, and pre- and post-test discussions. Posttest discussions should be followed up with additional social, psychological, and medical support and counselling. PMID- 15826433 TI - Detecting suicidal ideation in older patients: identifying risk factors within the general practice setting. AB - BACKGROUND: GPs are the most frequently accessed health professional among suicidal individuals in the community. AIM: To determine the prevalence of psychological distress and suicidal ideation among patients aged 60 years and older presenting to GPs, and the relationship between these variables in detecting patients who may be contemplating suicide. DESIGN OF STUDY: Cross sectional analysis of older patients presenting to Australian GPs between 2002 and 2003. SETTING: One thousand and sixty-one consecutive patients aged 60 years or over attending one of 54 randomly selected Western Australian GPs. METHOD: Prior to their medical consultation, patients completed a self-report questionnaire, which included questions about current suicidal ideation (Depressive Symptom Inventory Suicidality Subscale [DSI-SS]) and depression (Center for Epidemiological Studies Depression Scale [CES-D]). Patients' chief complaints were obtained from consultation summary sheets completed by their GP. RESULTS: Although only 5.1% of patients presented with psychological complaints, 5.8% acknowledged current suicidal ideation and 23.8% had clinically significant levels of depressive symptomatology. Suicidal ideation was associated with CES-D scores greater than 16 (odds ratio [OR] = 3.7, 95% confidence interval [CI] = 1.5 to 8.9), feelings of depression (OR = 7.7, 95% CI = 3.4 to 17.7), and previous suicide attempt (OR = 7.4, 95% CI = 2.7 to 20.2) in a logistic regression model, but not with poor self-perceived health, use of licit drugs (smoking, alcohol, and hypnotics), or type of presenting complaint at the time of assessment. CONCLUSIONS: Although older general practice patients tend to present for issues related to their physical health, approximately a quarter of this cohort also possess high levels of psychological distress, including current thoughts of suicide. Older patients who show any signs of depression or distress should be asked about psychological symptoms, including suicidal ideation. PMID- 15826434 TI - A cross-sectional survey of patients' beliefs about stress and their help-seeking behaviour. AB - BACKGROUND: Stress has become an increasingly common presentation in general practice. This may relate to an increase in stress in people's lives or a change in the meaning of stress and its conceptualisation as a legitimate problem for the GP. AIM: To explore patients' beliefs about stress, their association with help-seeking behaviour, and to examine differences by ethnic group. DESIGN OF STUDY: Cross-sectional survey of general practice patients attending to see their GP. SETTING: An inner-city London practice. METHOD: Consecutive general practice patients completed a questionnaire, which involved rating a series of symptoms for the extent to which they were associated with stress and describing their help-seeking behaviour. In total, 548 patients completed the questionnaire. Most patients described themselves as black Caribbean (n = 163), black African (n = 48), or white British (n = 187). RESULTS: The symptoms most frequently associated with stress were sleeping problems, feeling depressed, feeling panicky, having high blood pressure and feeling anxious; feeling ashamed, experiencing indigestion, having diarrhoea, feeling hot or cold, and suffering from constipation were least commonly associated to stress. This model of stress did not vary by ethnic group. Ethnic group differences were found for the association between the model of stress and help-seeking behaviour. Although white British patients consistently reported that the more a symptom was seen as indicative of stress, the more likely they would be to visit the doctor for that symptom, this association was not found for either black Caribbean or black African patients. CONCLUSIONS: The belief that stress-related symptoms are a legitimate problem for the GP is not universal and varies according to ethnic group. Stress is used by different patients in different ways and offers a variable pathway to the doctor. PMID- 15826435 TI - GP treatment decisions for patients with depression: an observational study. AB - BACKGROUND: GPs are prescribing more antidepressants than previously, but not in accordance with guidelines. The reasons why they prescribe are not well understood. AIM: To explore associations between GP treatment and severity of depression, patients' life difficulties, previous history of illness and treatment, and patient attitudes. DESIGN: Observational study in two phases, 3 years apart. SETTING: Seven practices in Southampton, UK. METHOD: Adult attenders who consented were screened for depression in the waiting room. After the consultation, the 17 participating GPs completed questionnaires on the perceived presence and severity of depression, patients' life difficulties, previous problems and treatment, patient attitudes towards antidepressants, and their treatment decisions. Patients returned postal questionnaires on sociodemographics, life events, physical health, and attitudes towards antidepressants. RESULTS: Of 694 patients screened in the two phases, the GPs rated 101 (15%) as depressed, acknowledged depression in 44 cases (6%), and offered treatment in 27 (4%), including antidepressants in 14 (2%). Offers of antidepressants were more likely in both phases where the GPs rated the depression as moderate rather than mild, and where they perceived a positive patient attitude to antidepressants. However, GP ratings of severity did not agree well with the validated screening instrument, and their assessments of patients' attitudes to treatment were only moderately related to patients' self reports. CONCLUSIONS: In line with current guidelines, GPs base prescribing decisions on the perceived severity of depression, taking patients' preferences into account, but they do not accurately identify which patients are likely to benefit from treatment. Better ways to assess depression severity and patient attitudes towards antidepressants are needed in order to target treatment more appropriately. PMID- 15826436 TI - The effect of deprivation, age and sex on NHS Direct call rates. AB - BACKGROUND: The use of primary care services in the UK is traditionally high in deprived areas. There has been little research into the effect of deprivation on the uptake of NHS Direct, a national nurse-led health helpline. AIM: To explore the impact of deprivation, age and sex on call rates to two NHS Direct sites. DESIGN OF STUDY: Ecological study. SETTING: West Yorkshire and West Midlands NHS Direct sites. METHOD: Details of NHS Direct calls between July 2001 and January 2002 were linked to electoral wards and the Indices of Multiple Deprivation for 2000. Age-standardised call rates were calculated for five deprivation levels. Using a negative binomial regression model, West Yorkshire call rates were analysed by age group, sex, deprivation level and geographical location. Rates were mapped by ward for West Yorkshire NHS Direct. RESULTS: Six-monthly call rates were highest for children under 5 years of age (130 per 1000 population). The ratio of female to male calls (all ages) was 1.30 (95% confidence interval [CI] = 1.27 to 1.33), this ratio being highest for the 15-44 year age group (P < 0.001). For both West Yorkshire and West Midlands NHS Direct, call rates (all ages combined) were highest in areas within the middle of the range of deprivation. West Yorkshire call rates about those under 5 years of age were lower in the most deprived areas than in the least deprived areas (< 1 year, P = 0.06; 1-4 years, P = 0.03). For adults aged 15-64 years, call rates were significantly higher in the most deprived areas (P < 0.001). CONCLUSION: This work supports previous research and shows that overall demand for NHS Direct is highest in areas where deprivation is at or just above the national average. Additionally, this study suggests that the effect of extreme deprivation appears to raise adult call rates but reduce rates of calls about children. PMID- 15826437 TI - UK childbirth delivery options in 2001-2002: alternatives to consultant unit booking and delivery. AB - BACKGROUND: Government policy advocates maternal choice in pregnancy care. Two key issues are place of birth and type of lead professional. Anecdotal evidence suggests there is variation in both these issues across the UK, but there has been no recent national assessment of whether maternal options are in line with government policy. AIM: To establish the range of women's childbirth delivery options, degree of midwife autonomy, and supporting training and governance mechanisms. DESIGN: Two postal questionnaires. SETTING: UK maternity units. METHOD: Questionnaires were sent to maternity services managers. MAIN OUTCOME MEASURES: number and type of units and births, transfers and care types; midwifery procedures; clinical governance and training activities. RESULTS: Completed questionnaires were received from 301 out of 308 (97.7%) units in 2002 and from 258 out of 309 (83.5%) units in 2001. Midwife-led care is available in 186 English (76.9%), 15 Welsh (78.9%), 18 Scottish (48.6%) and three Northern Ireland (30.0%) units. There are 73 (24.3%) stand-alone, 22 (7.3%) alongside, 127 (42.2%) integrated and 79 (26.2%) consultant units (for definitions of unit types, see main text), with a median 2215 hospital, 25 home and 210 midwife-led births. The median antenatal and labour transfers from midwife-led units are 25.5% (interquartile range [IQR] = 18.5-36.5%) and 18.0% (IQR = 13.4-24.8%) respectively; transfers are independent of distance to nearest consultant unit, country and unit type. CONCLUSIONS: Despite government policy promoting greater parental choice, this is not in evidence in many parts of the UK. The wide variations in home birth, midwife-led care and maternity-unit types merit further exploration. If more midwife-led units are to be established as a way of promoting parental choice and dealing with junior doctor rota problems, then such units must have adequate governance and training activities in place. PMID- 15826438 TI - Understanding of blood pressure by people with type 2 diabetes: a primary care focus group study. AB - BACKGROUND: For many people with type 2 diabetes most care is provided in primary care. While people with both diabetes and hypertension are at increased risk of complications, little is known about their understanding of blood pressure. AIM: To explore the understanding and beliefs about the importance of blood pressure held by people with type 2 diabetes. DESIGN OF STUDY: Framework analysis of qualitative research using focus groups. SETTING: Thirty-two participants were recruited from four general practices and a religious meeting group in Nottingham. Discussions took place in five community centres providing familiar surroundings for participants. METHOD: In order to get views expressed fully, white, Asian, and African-Caribbean participants met in five separate groups. Facilitators were fluent in the appropriate language and one member of the research team was present at all focus groups. RESULTS: Some participants, including those with raised blood pressure, were not aware of the increased importance of achieving good blood pressure control. No participants mentioned the increased risk of eye or kidney disease as a result of the combination of diabetes and raised blood pressure. Participants' perceptions regarding the control of blood sugar and blood pressure were different: blood sugar control was seen as their responsibility but blood pressure control was seen as the responsibility of the doctor. There was scepticism regarding the diagnosis of raised blood pressure, of targets and the management of blood pressure. There was also scepticism about the advice and education about diabetes given in primary care. CONCLUSIONS: People with type 2 diabetes require more knowledge of the increased risks they have from raised blood pressure, although this alone is unlikely to improve blood-pressure control. Strategies to increase the degree of control over and responsibility taken for the control of blood pressure need development and may require the specific development of participatory and negotiating skills among people with type 2 diabetes. Increasing the participation of these people in their own care will require doctors and nurses to work in a different way. PMID- 15826440 TI - Qualifications. PMID- 15826439 TI - Motivational interviewing: a systematic review and meta-analysis. AB - BACKGROUND: Motivational Interviewing is a well-known, scientifically tested method of counselling clients developed by Miller and Rollnick and viewed as a useful intervention strategy in the treatment of lifestyle problems and disease. AIM: To evaluate the effectiveness of motivational interviewing in different areas of disease and to identify factors shaping outcomes. DESIGN OF STUDY: A systematic review and meta-analysis of randomised controlled trials using motivational interviewing as the intervention. METHOD: After selection criteria a systematic literature search in 16 databases produced 72 randomised controlled trials the first of which was published in 1991. A quality assessment was made with a validated scale. A meta-analysis was performed as a generic inverse variance meta-analysis. RESULTS: Meta-analysis showed a significant effect (95% confidence interval) for motivational interviewing for combined effect estimates for body mass index, total blood cholesterol, systolic blood pressure, blood alcohol concentration and standard ethanol content, while combined effect estimates for cigarettes per day and for HbA(1c) were not significant. Motivational interviewing had a significant and clinically relevant effect in approximately three out of four studies, with an equal effect on physiological (72%) and psychological (75%) diseases. Psychologists and physicians obtained an effect in approximately 80% of the studies, while other healthcare providers obtained an effect in 46% of the studies. When using motivational interviewing in brief encounters of 15 minutes, 64% of the studies showed an effect. More than one encounter with the patient ensures the effectiveness of motivational interviewing. CONCLUSION: Motivational interviewing in a scientific setting outperforms traditional advice giving in the treatment of a broad range of behavioural problems and diseases. Large-scale studies are now needed to prove that motivational interviewing can be implemented into daily clinical work in primary and secondary health care. PMID- 15826441 TI - Failing asylum seekers. PMID- 15826442 TI - Corticosteroid injections: from bench to bedside? PMID- 15826443 TI - Gender-based inequalities. PMID- 15826444 TI - Teenage pregnancy: more than meets the eye. PMID- 15826445 TI - SAPC: scotching the myths. PMID- 15826446 TI - Drinking to, and gambling with, our health. PMID- 15826447 TI - Will GP revalidation make things better or worse? PMID- 15826453 TI - Don quixote at 400. PMID- 15826449 TI - What do we mean by health? PMID- 15826455 TI - Inequalities and elections (with reference to Robert Burns). PMID- 15826460 TI - Bariatric surgery is a cost-saving for the healthcare system. PMID- 15826461 TI - Health-related quality of life and symptoms of depression in extremely obese persons seeking bariatric surgery. AB - BACKGROUND: Extreme obesity, defined by a body mass index (BMI) > or =40 kg/m2, is associated with increased risk of depression and with impairments in healthrelated quality of life (HRQoL). This study examined the relationships among BMI, HRQoL, and symptoms of depression in persons with extreme obesity. METHOD: Participants were 306 patients who sought bariatric surgery (mean +/- SD age 43.8+/-9.4 years, BMI 52.8+/-9.3 kg/m2) and who completed questionnaires to assess symptoms of depression and HRQoL prior to surgery. We defined HRQoL impairment as a score > or =1 SD below national means. RESULTS: Impairments in HRQoL were common: >40% of participants scored in the impaired ranges of physical functioning, physical role limitations, and bodily pain. Results of analyses of variance (ANOVAs) indicated that impairments in HRQoL were significantly related to symptoms of depression. Patients with impaired HRQoL scored above the cut-off point for clinically significant depression, while those without such impairment scored below that cut-off, regardless of BMI. The contribution of BMI to depression was not significant in any ANOVA. CONCLUSION: HRQoL is more strongly and more directly related to symptoms of depression than is BMI. These findings highlight the need to assess HRQoL in patients with extreme obesity and suggest that interdisciplinary strategies to address HRQoL impairments may be beneficial in this population. PMID- 15826462 TI - Predictors of nonalcoholic steatohepatitis and advanced fibrosis in morbidly obese patients. AB - BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is a common form of chronic liver disease in the United States. It is commonly associated with the components of the metabolic syndrome including obesity. From the spectrum of NAFLD, only patients with nonalcoholic steatohepatitis (NASH) have been convincingly shown to have a potential for progression to cirrhosis. We report the prevalence of NAFLD and NASH as well as predictors of NASH and advanced fibrosis in morbidly obese patients. METHODS: 212 consecutive patients who underwent bariatric surgery were enrolled in the study. A liver biopsy was performed at the time of the surgery. Causes of chronic liver disease other than NAFLD were excluded by clinical and laboratory evaluation. RESULTS: The prevalence of NAFLD was 93%. Of those with NAFLD, 26% had NASH. 17 patients (9%) had advanced fibrosis (i.e., bridging fibrosis or cirrhosis). Male gender, AST, and type 2 diabetes mellitus were independently associated with NASH. Waistto-hip ratio, AST, and focal hepatocyte necrosis on liver biopsy were independently associated with advanced fibrosis. Interestingly, while AST was associated with NASH and advanced fibrosis, the majority of the patients with either NASH or advanced fibrosis had normal AST. CONCLUSIONS: NAFLD and NASH are very common in morbidly obese patients undergoing bariatric surgery. Features associated with the metabolic syndrome and liver cell injury are independently associated with either NASH or advanced fibrosis. PMID- 15826463 TI - Reoperative laparoscopic Roux-en-Y gastric bypass: an experience with 49 cases. AB - BACKGROUND: Long-term complications leading to reoperation after primary bariatric surgery are not uncommon. Reoperations are particularly challenging because of tissue scarring and adhesions related to the first operation. Reoperations must address the complication(s) related to the scarring and, at the same time, prevent weight regain that would inevitably occur after simple reversal. Conversion to Roux-en-Y gastric bypass (RYGBP) has repeatedly been demonstrated to be the procedure of choice in most situations. It has traditionally been performed through an open approach. Our aim is to describe our experience with the laparoscopic approach in reoperations to RYGBP over the past 5 years. METHODS: All patients undergoing laparoscopic RYGBP as a reoperation were included in this study. Patients with multiple previous operations or patients with band erosion after gastric banding were submitted to laparotomy. Data were collected prospectively. RESULTS: Between June 1999 and August 2004, 49 patients (44 women, 5 men) underwent laparoscopic reoperative RYGBP. The first operation was gastric banding in 32 and vertical banded gastroplasty in 15. The mean duration of the reoperation was 195 minutes. No conversion to open was necessary. Overall morbidity was 20%, with major complications in 2 patients (4%). Weight loss, or weight maintenance, was satisfactory, with a BMI <35 kg/m2 up to 4 years in close to 75% of the patients. CONCLUSIONS: Laparoscopic RYGBP can be safely performed as a reoperation in selected patients provided that the surgical expertise is available. These procedures are clearly more difficult than primary operations, as reflected by the long operative time. Overall morbidity and mortality, however, are not different. Long-term results regarding weight loss or weight maintenance are highly satisfactory, and comparable to those obtained after laparoscopic RYGBP as a primary operation. PMID- 15826464 TI - Unchanged hypovitaminosis D and secondary hyperparathyroidism in morbid obesity after bariatric surgery. AB - BACKGROUND: Morbidly obese patients have been reported to present with vitamin D insufficiency and secondary hyperparathyroidism. We assessed whether bariatric surgery alters the 25-hydroxyvitamin D (calcidiol) and intact parathyroid hormone (iPTH) levels in patients presenting with morbid obesity. METHODS: A cross sectional survey was conducted on 144 patients of whom 80 had not undergone bariatric surgery, while 64 had bariatric surgery at a mean of 36 months previously. Calcidiol levels were defined as being normal (>50 nmol/L), insufficient (2550 nmol/L) and deficient (<25 nmol/L). Mild secondary hyperparathyroidism was defined as iPTH >7.3 pmol/L with simultaneous normal values for creatinine, calcium and phosphorus. RESULTS: 80% of the patients presented low vitamin D levels and mild secondary hyperparathyroidism. Previous surgery or the presence of diabetes did not influence calcidiol levels. Corrected serum calcium, phosphorus, alkaline phosphatase, iPTH and Calcidiol were similar between subjects with and without surgery. CONCLUSIONS: Vitamin D deficient states with secondary hyperparathyroidism in the morbidly obese precede and are not significantly affected by bariatric surgery. Hypovitaminosis D with secondary hyperparathyroidism due to low calcidiol bio-availability should be added to the crowded list of sequelae of morbid obesity. While further studies are warranted, it seems advisable to support vitamin D supplementation in the morbidly obese population. PMID- 15826465 TI - Continuous positive airway pressure (CPAP) changes in bariatric surgery patients undergoing rapid weight loss. AB - BACKGROUND: Obstructive sleep apnea (OSA) is a common condition in morbidly obese patients, with the reported prevalence ranging from 12-78%. There is increasing recognition of the need to diagnose and treat/manage OSA both preoperatively and postoperatively. Nasal CPAP is the preferred treatment of OSA; however, weight loss is associated with a reduction in required pressures. We evaluated the CPAP pressure requirements in a group of patients undergoing rapid weight loss following Roux-en-Y gastric bypass. METHODS: 15 patients who had been diagnosed with OSA before surgery were retrospectively evaluated. All patients had demonstrated compliance on home CPAP therapy, were minimally 3 months post surgery and had follow-up reports that their CPAP was less effective. We obtained data on age, sex, weight, BMI, and apnea/hypopnea index (AHI). Optimal CPAP pressure was obtained initially through attended in-laboratory complex polysomnography. Follow-up CPAP pressure was obtained using an auto-titrating PAP device at home. These data were used to evaluate the pressure changes that accompanied weight loss. RESULTS: This group of patients had lost an average of 44.5 +/- 19.4 kg. Four patients had achieved their goal weight. Their starting CPAP pressures averaged 11 +/- 3.0 cm H2O, with a range of 7-18 cm H2O. Follow-up CPAP pressures averaged 9 +/- 2.7 cm H2O, with a range of 4-12 cm H2O, representing an overall reduction of 18%. The subgroup of patients who had achieved goal weight had a pressure reduction of 22% (9 +/- 2.0 to 7 +/- 1.0 cm H2O). CONCLUSION: CPAP pressure requirements change considerably in bariatric surgery patients undergoing rapid weight loss. Auto-titrating PAP devices have promise for facilitating the management of CPAP therapy during this time. Consideration should also be given to the use of autotitrating PAP units as the treatment of choice in these patients. PMID- 15826466 TI - Endoscopy after Roux-en-Y gastric bypass: a community hospital experience. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGBP) is a common surgical intervention for morbid obesity. Postoperative GI symptoms are common. This study reports the endoscopic findings in symptomatic patients. METHODS: Patients who developed GI symptoms after RYGBP at a single community hospital were referred for endoscopic evaluation. Standard endoscopic procedures using standard endoscopic equipment were used. RESULTS: From April 2002 to April 2004, 23 out of 200 patients underwent 35 endoscopic procedures. All patients complained of some degree of epigastric pain, nausea and vomiting regardless of endoscopic findings. The most common endoscopic finding was ulcer disease (12 patients - 52%). Other findings included normal postoperative anatomy (7 patients - 30%), anastomotic stricture (1 patient - 4.3%), obstructed biliopancreatic limb (1 patient - 4.3%), acute gastric pouch bleed (1 patient - 4.3%), anastomotic rupture/dehiscence (1 patient - 4.3%). H. pylori was not detected in any patient. CONCLUSIONS: In patients who have had RYGBP, symptoms were a poor predictor of endoscopic pathology. Ulcer disease was the most common endoscopic finding. These ulcers were not associated with H. pylori. All ulcers responded well to oral proton pump inhibitors (PPI) and sucralfate therapy. The community gastroenterologist should be acquainted with the typical post-surgical anatomy and possible endoscopic intervention for RYGBP patients. PMID- 15826467 TI - Patient characteristics impacting excess weight loss following laparoscopic adjustable gastric banding. AB - BACKGROUND: Weight loss is more variable after laparoscopic adjustable gastric banding (LAGB) than after gastric bypass. Subgroup analysis of patients may offer insight into this variability. The aim of our study was to identify preoperative factors that predict outcome. METHODS: Demographics, co-morbid conditions and follow-up weight were collected for our 1st 200 LapBand patients. Linear regression determined average %EWL. Logistic regression analysis identified factors that impacted %EWL. RESULT: 200 patients returned for 778 follow-up visits. Median age was 44 years (21-72) and median BMI 45 kg/m2 (31-76). 140 (80%) were women. Average %EWL was y % = 0.007 %/day (days since surgery) + 0.12% (correlation coef. 0.4823; P<0.001). %EWL at 1 year was 37%. The best-fit logistic regression model found 7 factors that significantly changed the odds of achieving average %EWL. Older patients, diabetic patients and patients with COPD had greater odds of above average %EWL. Female patients, patients with larger BMIs, asthmatic patients and patients with hypertension had increased odds of below average %EWL. CONCLUSION: Specific patient characteristics and comorbid conditions significantly altered the odds of achieving satisfactory %EWL following gastric banding. PMID- 15826468 TI - Results after laparoscopic adjustable gastric banding in patients over 55 years of age. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) has become the most popular bariatric intervention in Europe. International guidelines recommended age limits for bariatric surgery of 18-60 years. The aim of this study was to evaluate the immediate results in morbidly obese patients >55 years old, treated with LAGB. METHODS: Between January 1996 and January 2004, 350 patients underwent LAGB. 24 (6.8%) were >55 years old (Group A), mean age 58.6+/-3.3 years, mean preoperative BMI 42.3+/-4.5 kg/m2. A comparative randomized analysis with 24 patients younger than age 55 years was performed (Group B: mean age 41.2+/-9.6 years, mean BMI 42.1+/-3.6 kg/m2). Baseline clinical features, operative parameters and postoperative results were evaluated. RESULTS: No perioperative complications were recorded. Conversion rate and mortality were nil. Major postoperative complications occurred in 2 patients (8.3%) from Group A (1 intragastric prosthesis migration, 1 pouch dilatation) and 2 patients (8.3%) from Group B (intragastric migrations). Reoperation was needed in 3 cases, and one erosion (Group B) is on the waiting list for removal. Minor complications: 1 port infection in each group required ambulatory port substitution; 1 intraperitoneal portcatheter disconnection (Group B) was successfully treated laparoscopically. Mean follow-up was 31.7 months (Group A) and 33.0 months (Group B). Mean postoperative BMI at 12 and 24 months was 35.9+/-4.2 and 33.8+/-4.9 for Group A, and 33.8+/-4.6 and 33.2+/-6.0 for Group B. CONCLUSION: There have been no significant differences in results between the 2 groups. LAGB has been safe and effective in patients >55 years old. PMID- 15826469 TI - Obstructive symptoms associated with the 9.75-cm Lap-Band in the first 24 hours using the pars flaccida approach. AB - BACKGROUND: For some patients, especially those with a higher BMI, a non selective Lap-Band placement using the pars flaccida approach with application of the small-diameter bands (9.75 and 10 cm) may be too tight or may require significant gastroesophageal junction dissection and thinning. In such a case, the major perioperative complication is acute obstruction immediately after surgery. We review the etiology of obstructive complications that present postoperatively in the first 24 hours. CASE REPORTS: Acute postoperative stoma obstruction (esophageal outlet stenosis) was observed in 5 patients who underwent 9.75-cm Lap-Band placement for morbid obesity. 2 of these patients had a postoperative upper GI series showing a misplaced band with gastric slippage, and repeat operation was required. 3 patients had gastric obstruction without slippage. Of the latter, 1 patient insisted that the band be removed rather than being replaced with a longer one, and the remaining 2 were managed with conservative treatment, involving extended hospitalization until the edema subsided and the patient slowly regained the ability to swallow. CONCLUSION: Obstructive symptoms associated with the Lap-Band using the pars flaccida approach can be addressed conservatively in most patients or by minimally invasive surgery; however we believe that routine use of the 11-cm Lap-Band for the pars flaccida approach could easily prevent this early complication. PMID- 15826470 TI - Port complications following laparoscopic adjustable gastric banding for morbid obesity. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) has gained widespread acceptance. However, the technique has problems intrinsic to the material wear and tear around the port and connecting tubing that can lead to failure. Port complications are considered to be minor; however, few studies have analyzed them, and the optimal technique of port implantation and management has not been elucidated. METHODS: All patients who suffered from complications involving the tubing or access-port were included in this study. Their complaints, imaging studies, operative reports and hospitalization files were retrospectively reviewed. RESULTS: 1,272 of the patients were available for a mean follow-up period of 37 months. During this time, 91 patients (7.1%) experienced port complications that required 103 revisional operations. Of these patients, 62 had system leaks, 19 infectious problems, and 10 miscellaneous problems requiring operative correction. Overall port problems led to band removal in 6 patients, and replacement in 1 patient. CONCLUSION: Access-port complications after the Lap Band procedure are among the most common and annoying ones, and can render the device susceptible to failure. Careful surgical technique and routine use of radiologic guidance for band adjustments are the keys to avoiding complications. PMID- 15826471 TI - Invited commentary: port and tubing complications. PMID- 15826472 TI - Comparison of changes in lipid profile after bilio-intestinal bypass and gastric banding in patients with morbid obesity. AB - BACKGROUND: The presence of hypercholesterolemia is currently not considered a selection criteria for performing gastric restrictive or diversionary bariatric surgery. METHODS: We prospectively investigated the effects of the bilio intestinal bypass (BI-bypass) with a wide cholecysto-jejunal anastomosis and of adjustable gastric banding (AGB) on blood lipid concentrations in obese patients. To clarify the mechanism of the hypocholesterolemic effect of the BI-bypass, daily fecal sterol excretion was measured by gas-liquid chromatography (GLC). RESULTS: At 1 year after BI-bypass compared to baseline, the hypercholesterolemic (n=18) and the normocholesterolemic (n=19) patients significantly reduced total ( 38% and -27%, respectively), LDL (-47% and -24%, respectively) and HDL (-11% and 13%, respectively) cholesterol and total / HDL cholesterol ratio (-25% and -13%, respectively). At 1 year after AGB, the total / HDL cholesterol ratio was significantly decreased (-11%) compared to baseline in hypercholesterolemic (n=12) but not in normocholesterolemic (n=6) patients, while total and LDL cholesterol were not affected in both groups. At 3 years after BI-bypass compared to baseline, the hypercholesterolemic (n=9) and the normocholesterolemic (n=11) patients significantly reduced total (-43% and -28%, respectively) and LDL (-53% and -29%, respectively) cholesterol and total / HDL cholesterol ratio (-38% and 21%, respectively). The BI-bypass induced a significant (P <0.005; n=7) 6-fold increase in mean fecal cholesterol output. CONCLUSIONS: The BI-bypass but not the AGB leads to a persistent and marked beneficial effect on blood LDL cholesterol associated with an increased cholesterol fecal output. BI-bypass but not AGB is indicated in morbidly obese patients with hypercholesterolemia. PMID- 15826473 TI - The effect of surgical weight reduction on functional status in morbidly obese patients with low back pain. AB - BACKGROUND: Although low back (LBP) pain is not a life-threatening disease, it is a source of significant discomfort and disability and accounts for work absences. It has been shown previously that morbid obesity is associated with increased frequency of LBP and that surgical weight loss improves the symptomatology. However, there are no studies to quantitatively assess the exact degree of functional disability caused by severe obesity and the degree of improvement of LBP that follows weight loss from bariatric surgery. METHODS: 29 morbidly obese candidates for bariatric surgery with LBP, weight 132.5+/-27 (mean+/-SD) kg and BMI 47.2+/-8.8 kg/m2 were examined for their functional status using psychometric instruments specifically designed to objectively assess the patients' complaints. The preoperative scores were measured by a) visual analogue scales (VAS1, VAS2, VAS3), b) Roland-Morris disability questionnaire, c) Oswestry LBP disability questionnaire, and d) Waddell disability index, and were compared with the scores obtained by the same instruments 2 years after vertical banded gastroplasty. RESULTS: The postoperative weight (92.3+/-19 kg) and BMI (32.9+/-6.3 kg/m2) of the 29 patients were significantly reduced (P<0.001). The improved functional disability scores were statistically significant: a) VAS1 1.59+/-1.86 (mean+/-SD) vs 0.32+/-0.64, P<0.001; b) VAS2 5.5+/-1.97 vs 2.14+/-1.88, P<0.001; c) VAS3 0.77+/-1.11 vs 0.09+/-0.29, P=0.006, d) Roland-Morris 7.89+/-5.11 vs 1.89+/-2.13, P<0.001; e) Oswestry 21.22+/-15.63 vs 5.61+/-7.51, P<0.001; f) Waddell 2.81+/ 1.37 vs 0.56+/-0.72, P<0.001. CONCLUSIONS: Surgical weight loss significantly improves the degree of functional disability of morbidly obese patients suffering from LBP. PMID- 15826474 TI - Adiponectin expression and adipose tissue lipolytic activity in lean and obese women. AB - BACKGROUND: The authors evaluated whether adipose adiponectin expression is related to adipose tissue lipolytic activity and fatty acid oxidation rate in lean and obese women. METHODS: The study consisted of 60 adult females distributed in a wide range of adiposity (BMI 24.0-53.4 kg/m2). Body composition was estimated by bioelectrical impedance. Respiratory quotient was measured by open-circuit indirect calorimetry. RT-PCR assays were performed to measure TNFalpha and adiponectin expression in subcutaneous adipose tissue biopsies. Lipolysis studies were performed in fresh tissue samples obtained from subcutaneous abdominal depots during bariatric surgery in 19 morbidly obese females or by an incisional biopsy or during abdominal elective surgery in normal weight and obese females. Glycerol release was measured with a colorimetric endpoint method. RESULTS: Patients with a higher degree of adiposity showed lower adipose tissue adiponectin expression and hormone-sensitive lipase (HSL) activity than women in the low range of adiposity. HSL activity was positively related to adiponectin expression. No relationship was observed between adiponectin and TNFalpha subcutaneous adipose tissue expression. The positive relationship between respiratory quotient and adiponectin expression was in the limit of statistical significance (P=0.05). Percentage of body fat and mRNA adiponectin explained 26% of the variance of the adipose tissue HSL activity. CONCLUSIONS: Low adipose tissue adiponectin expression observed in obese people may contribute to the progression of obesity and its co-morbidities by modulating hormone sensitive lipase activity and fatty acid oxidation. PMID- 15826475 TI - Changes in glucagon-like peptide-1 (GLP-1) secretion after biliopancreatic diversion or vertical banded gastroplasty in obese subjects. AB - BACKGROUND: Bariatric operations promote weight loss and improve glucose homeostasis. Glucagon-like peptide-1 (GLP-1) is considered as a possible mediator of the antidiabetic effects of such operations. METHODS: The present study aimed to gain information on the time course for changes in glucose tolerance, as well as insulin, glucagon and GLP-1 secretion, during an oral glucose tolerance test (OGTT), in 31 obese patients examined 1, 3 and 6 months after Larrad's biliopancreatic diversion (BPD) or 6 months after vertical banded gastroplasty (VBG). RESULTS: A time-related progressive decrease in body weight coincided with lowering of plasma triglycerides, decrease of basal plasma glucose and its incremental area during OGTT, and reduction of basal plasma insulin together with an increase of its incremental area. The time-related decrease of plasma glucagon during OGTT was comparable before and after surgery. Both the basal plasma GLP-1 concentration and its incremental area during the OGTT increased strikingly after surgery, a steady-state situation being reached 3 months after surgery. The most striking differences between the somewhat older and less glucose-tolerant subjects of VBG compared to BPD after surgery, consisted in a decrease in cholesterol and LDL only observed in BPD and a much more pronounced increase in basal and incremental plasma GLP-1 in BPD. GLP-1, like glucagon, increased lipolysis, but failed to duplicate the lipogenetic action of insulin in isolated adipocytes obtained at the time of surgery. CONCLUSION: These findings support the postulated role of GLP-1, secreted by the hindgut, as a key mediator of the antidiabetic effects of bariatric operations. PMID- 15826476 TI - One-anastomosis gastric bypass by laparoscopy: results of the first 209 patients. AB - BACKGROUND: One-Anastomosis Gastric Bypass (OAGB) by laparoscopy consists of constructing a divided 25-ml (estimated) gastric pouch between the esophago gastric junction and the crow's foot level, parallel to the lesser curvature, which is anastomosed latero-laterally to a jejunal loop 200 cm distal to the ligament of Treitz. METHODS: The results of our first 209 OAGB patients operated from July 2002 to June 2004 are reported. Mean age was 41 years (14-66), BMI 48 (39-86) and mean excess body weight 66 kg (35-220). In 144 patients, OAGB was the only operation performed, and in 61 patients it was accompanied by other surgery (18 cholecystectomies, 5 incisional hernia repairs, and 38 adhesiolysis), and in 4 patients a restrictive bariatric operation had been performed previously. RESULTS: 2 patients (0.9%) were converted to open surgery due to uncontrollable bleeding. 3 patients (1.4%) needed re-operation in the immediate postoperative period. 5 patients (2.3%) needed prolonged hospital stay due to acute pancreatitis in 1 and anastomotic leakage in 4, all resolving with conservative treatment. 2 patients died (0.9%), 1 from fulminant pulmonary thromboembolism and 1 from nosocomial pneumonia. Long-term complications have occurred in only 2 patients who developed clinically significant iron-deficiency anemia. Mean excess weight loss was 75% after 1 year and >80% at 2 years. CONCLUSION: OAGB is a simple, safe and effective operation with less perioperative risk than conventional gastric bypass, quicker return to normal activities, and better quality of life. PMID- 15826477 TI - Development of acute gouty attack in the morbidly obese population after bariatric surgery. AB - BACKGROUND: Gout is associated with increased body weight. We evaluated the prevalence of gout and acute gouty attacks in the morbidly obese population who underwent bariatric surgery. METHODS: The medical records and operative reports of 1,240 patients who underwent bariatric surgery were reviewed retrospectively for weight parameters, BMI, weight loss, medical history of gout, and onset of acute gouty attacks. RESULTS: Of the 1,240 patients, 5 (0.4%) had been previously diagnosed with gout. 2 of these 5 had acute attacks during the postoperative period, and responded successfully to intravenous colchicine. CONCLUSION: Although rare, gout must be considered a co-morbid illness in obese and morbidly obese patients. Surgeons should be familiar with the signs and symptoms of attacks in the postoperative period, and be knowledgeable in the management. PMID- 15826478 TI - The biliopancreatic diversion with the duodenal switch: results beyond 10 years. AB - The BPD/DS, if properly performed, has the best long-term weight loss of any bariatric operation. It is easy to reverse or revise, has the least marginal ulcers, cures the highest percentage of co-morbidities, has the least failures, and permits normal although smaller meals. It is our opinion that the BPD/DS should be considered as the gold standard bariatric operation. PMID- 15826479 TI - Empowerment and self-management after weight loss surgery. AB - The bariatric surgery health-care professional team are often frustrated and discouraged when patients are unwilling or unable to follow their advice to achieve ideal outcomes after obesity surgery. The acute care model that suffices for other types of surgery is inadequate after a surgery that requires chronic life-long behavioral changes. Practical interventions that facilitate collaborative relationships and foster patient-centered practices are the key to giving up feeling responsible for the choices that patients make, by being responsible to them, and achieving better outcomes. PMID- 15826480 TI - Gastric cancer after Roux-en-Y gastric bypass. AB - Roux-en-Y gastric bypass (RYGBP) is one of the most commonly performed surgical procedures for morbid obesity. Several complications that may develop in the short- and long-term have been reported. We present a patient who presented with cancer in the bypassed stomach 8 years after RYGBP. Although the development of this lesion is rare and only a few cases have been reported, there are aspects worthy of discussion. Several monitoring, diagnostic and therapeutic alternatives are analyzed. PMID- 15826481 TI - Successful treatment of gastric fistula following rhabdomyolysis after vertical banded gastroplasty. AB - Vertical banded gastroplasty is a gastric restrictive operation which has been performed with very satisfactory results in our department. We present a 46 year old male with BMI 48, who experienced rhabdomyolsis after a VBG operation, complicated by perforation of an upper pouch ulcer and subsequent gastric fistula. Cardiac and renal failure occurred, necessitating intensive therapy with catecholamines, diuretics and hemodialysis. The patient underwent multiple operations, drainage of a retroperitoneal abscess, suture of a perforated ulcer, and gastric decompression by a gastrostomy. Prolonged treatment including TPN, drainage, broad spectrum antibiotics, skin and would protection and jejunostomy feeding, were necessary to obtain an eventual successful outcome. This case demonstrates that unexpected surgical complications may occur in morbidly obese patients and how difficult and long the management of these may be. Rhabdomyolsis is a potentially life-threatening complication of bariatric surgery, and careful postoperative observation of the patient is mandatory. PMID- 15826482 TI - Severe hypocalcemia following total thyroidectomy after biliopancreatic diversion. AB - Patients undergoing malabsoprtive operations for bariatric surgery are prone to disturbances of bone metabolism, but this does not commonly lead to clinical symptoms. We present a morbidly obese patient who had undergone the biliopancreatic diversion of Larrad, and presented clinical symptoms of severe hypocalcemia and tetany after total thyroidectomy. Very high doses of i.v. calcium and calcitriol and 10 days of hospitalization were required to control the symptoms and correct plasma levels. The physiological aspects that contribute to this situation are discussed. PMID- 15826483 TI - Simultaneous paraesophageal hernia repair and gastric banding. AB - The presence of a hiatal hernia is generally considered a contraindication to gastric banding in the morbidly obese, despite recent reports indicating favorable outcomes following simultaneous repair of sliding hernias and laparoscopic adjustable gastric banding (LAGB). A 66-year-old woman weighing 120 kg (BMI 45) with arterial hypertension and gastroesophageal reflux-related chronic obstructive pulmonary disease underwent repair of a large paraesophageal hernia and LAGB. At 40 months followup, the patient had lost 44% excess body weight (BMI 36) and had no complaints of heartburn, regurgitation or dysphagia. She was no longer hypertensive and her pulmonary condition had improved significantly. Barium swallow at 30 months showed normal anatomy and positioning of the band. Because other minimally traumatic surgical options are lacking, the author believes morbidly obese patients with hiatal hernia should not be denied the advantages of LAGB. Adequate weight reduction, resolution of gastroesophageal reflux and other co-morbidities can be expected if an appropriate surgical technique is used. PMID- 15826484 TI - A minimally invasive solution for necrotic fundus following slipped adjustable gastric band. AB - The laparoscopic adjustable gastric band (LAGB) is widely used for treatment of morbid obesity. Band slippage is a well known long-term complication. Herein, we present a patient with band slippage who developed strangulation of the fundus with irreversible necrosis. The chosen surgical solution was minimally invasive. A diagnostic laparoscopy and consequently resection of the necrotic fundus was performed. A postoperative symptomatic left pleural effusion resolved after chest tube insertion. One week later, the patient was discharged with satisfactory results. PMID- 15826485 TI - Non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma. AB - Non-alcoholic fatty liver disease (NAFLD) is characterized by an excessive accumulation of fatty acids and triglycerides within the cytoplasm of the hepatocytes of non-alcohol users. The natural history varies according to the initial histological diagnosis. A current consideration is that cryptogenic cirrhosis may be representative of a late stage of non-alcoholic steatohepatitis (NASH), which has lost its features of necroinflammatory activity and steatosis in up to 80% of patients. Since NASH is able to progress to cirrhosis, hepatocellular carcinoma (HCC) development may be an end-stage of this disease. We report below two clinical cases of patients diagnosed with NASH who developed HCC. The relationship between NAFLD and HCC is reviewed. PMID- 15826486 TI - Rapid resolution of diabetes after gastric bypass. PMID- 15826489 TI - Overview of innate immunity in Drosophila. AB - Drosophila protects itself from infection by microbial organisms by means of its pivotal defense, the so-called innate immunity system. This is its sole defense as it lacks an adaptive immunity system such as is found in mammals. The strong conservation of innate immunity systems in organisms from Drosophila to mammals, and the ease with which Drosophila can be manipulated genetically, makes this fly a good model system for investigating the mechanisms of virulence of a number of medically important pathogens. Potentially damaging endogenous and/or exogenous challenges sensed by specific receptors initiate signals via the Toll and/or Imd signaling pathways. These in turn activate the transcription factors Dorsal, Dorsal-related immune factor (Dif) and Relish, culminating in transcription of genes involved in the production of antimicrobial peptides, melanization, phagocytosis, and the cytoskeletal rearrangement required for appropriate responses. Clarifying the regulatory interactions between the various pathways involved is very important for understanding the specificity and termination mechanism of the immune response. PMID- 15826490 TI - Recent advances in the innate immunity of invertebrate animals. AB - Invertebrate animals, which lack adaptive immune systems, have developed other systems of biological host defense, so called innate immunity, that respond to common antigens on the cell surfaces of potential pathogens. During the past two decades, the molecular structures and functions of various defense components that participated in innate immune systems have been established in Arthropoda, such as, insects, the horseshoe crab, freshwater crayfish, and the protochordata ascidian. These defense molecules include phenoloxidases, clotting factors, complement factors, lectins, protease inhibitors, antimicrobial peptides, Toll receptors, and other humoral factors found mainly in hemolymph plasma and hemocytes. These components, which together compose the innate immune system, defend invertebrate from invading bacterial, fungal, and viral pathogens. This review describes the present status of our knowledge concerning such defensive molecules in invertebrates. PMID- 15826491 TI - Gene expression profiles of HeLa Cells impacted by hepatitis C virus non structural protein NS4B. AB - By a cDNA array representing 2308 signal transduction-related genes, we studied the expression profiles of HeLa cells stably transfected by Hepatitis C virus nonstructural protein 4B (HCV-NS4B). The alterations of the expression of four genes were confirmed by real-time quantitative RTPCR; and the aldo-keto reductase family 1, member C1 (AKR1C1) enzyme activity was detected in HCV-NS4B transiently transfected HeLa cells and Huh-7, a human hepatoma cell line. Of the 2,308 genes we examined, 34 were up-regulated and 56 were down-regulated. These 90 genes involved oncogenes, tumor suppressors, cell receptors, complements, adhesions, transcription and translation, cytoskeleton and cellular stress. The expression profiling suggested that multiple regulatory pathways were affected by HCV-NS4B directly or indirectly. And since these genes are related to carcinogenesis, host defense system and cell homeostatic mechanism, we can conclude that HCV-NS4B could play some important roles in the pathogenesis mechanism of HCV. PMID- 15826492 TI - Cloning and characterization of a gene encoding 22 kDa functional protein of bacteriophage MB78. AB - Functional protein of MB78 bacteriophage having apparent molecular weight of 22 kDa is expressed from 1.7 kb HindIII G fragment. The nucleotide sequence of this fragment showed two open reading frames of 222 and 196 codons in tail-to-tail orientation separated by a 62-nucleotide intercistronic region. The ORF of 22 kDa protein is present in opposite orientation, i.e. in the complementary strand, preceded by a strong ribosomal binding site and a promoter sequence. Another ORF started from the beginning of the fragment whose promoter region and translational start site lies in the 0.45kb HincII U fragment which is located next to the HindIII G fragment, that has the sequence for DNA bending. 3'end of the fragment has high sequence homology to the EaA and EaI proteins of bacteriophage P22, a close relative of MB78 phage. PMID- 15826493 TI - Differential expression and stability of endogenous nuclear factor E2-related factor 2 (Nrf2) by natural chemopreventive compounds in HepG2 human hepatoma cells. AB - Nuclear factor-E2-related factor 2 (Nrf2) is known as a key regulator of ARE mediated gene expression and the induction of Phase II detoxifying enzymes and antioxidant enzymes, which is also a common property of many chemopreventive agents. In the present study, we investigated the regulatory role of different chemopreventive agents including sulforaphane (SUL), allyl isothiocyanate (AITC), indole-3-carbinol (I3C), and parthenolide (PTL), in the expression and degradation of Nrf2 and the induction of the antioxidant enzyme HO-1. SUL strongly induced Nrf2 protein expression and ARE-mediated transcription activation, retarded degradation of Nrf2 through inhibiting Keap1, and thereby activating the transcriptional expression of HO-1. AITC was also a potent inducer of Nrf2 protein expression, ARE-reporter gene and HO-1 but had little effect on delaying the degradation of Nrf2 protein. Although PTL and I3C could induce AREreporter gene expression and Nrf2 to some extent, they were not as potent as SUL and AITC. However, PTL dramatically induced the HO-1 expression, which was comparable to SUL, while I3C had no effect. In addition, when treated with SUL and PTL, inhibition of proteasome by MG132 did not cause additional accumulation of Nrf2, suggesting the involvement of other degradation mechanism(s) in the presence of these compounds such as SUL and PTL. In summary, the results of our current study indicated that different chemopreventive compounds have different regulatory properties on the accumulation and degradation of Nrf2 as well as the induction of cellular antioxidant enzyme HO-1. PMID- 15826494 TI - Comparative study of enzyme activity and stability of bovine and human plasmins in electrophoretic reagents, beta-mercaptoethanol, DTT, SDS, Triton X-100, and urea. AB - Effects of common electrophoretic reagents, reducing agents (beta-mercaptoethanol [BME] and DTT), denaturants (SDS and urea), and non-ionic detergent (Triton X 100), on the activity and stability of bovine plasmin (b-pln) and human plasmin (h-pln) were compared. In the presence of 0.1% SDS (w/v), all reagents completely inhibited two plns, whereas SDS (1%) and urea (1 M) denatured plns recovered their activities after removal of SDS by treatment of 2.5% Triton X-100 (v/v). However, reducing agents (0.1 M of BME and DTT) treated plns did not restore their activities. Based on a fibrin zymogram gel, five (from b-pln) and four (from h-pln) active fragments were resolved. Two plns exhibited unusual stability in concentrated SDS and Triton X-100 (final 10%) and urea (final 6 M) solutions. Two bands, heavy chain-2 (HC-2) and cleaved heavy chain-2 (CHC-2), of b-pln were completely inhibited in 0.5% SDS or 3 M urea, whereas no significant difference was found in h-pln. Interestingly, 50 kDa (cleaved heavy chain-1, CHC-1) of b-pln and two fragments, 26 kDa (light chain, LC) and 29 kDa (microplasmin, MP), of h pln were increased by SDS in a concentration dependent manner. We also found that the inhibition of SDS against both plns was reversible. PMID- 15826495 TI - Genetic heterogeneity of the tropical abalone (Haliotis asinina) revealed by RAPD and microsatellite analyses. AB - Genetic heterogeneity of the tropical abalone, Haliotis asinina was examined using randomly amplified polymorphic DNA (RAPD) and microsatellite analyses. One hundred and thirteen polymorphic RAPD fragments were generated. The percentage of polymorphic bands of H. asinina across overall primers was 85.20%. The average genetic distance of natural samples within the Gulf of Thailand (HACAME and HASAME) was 0.0219. Larger distance was observed when those samples were compared with HATRAW from the Andaman Sea (0.2309 and 0.2314). Geographic heterogeneity and F(ST) analyses revealed population differentiation between H. asinina from the Gulf of Thailand and the Andaman Sea (p < 0.0001). Three microsatellite loci (CUHas1, CUHas4 and CUHas5) indicated relatively high genetic diversity in H. asinina (total number of alleles = 26, 5, 23 and observed heterozygosity = 0.84, 0.42 and 0.33, respectively). Significant population differentiation was also found between samples from different coastal regions (p < 0.0001). Therefore, the gene pool of natural H. asinina in coastal Thai waters can be genetically divided to 2 different populations; the Gulf of Thailand (A) and the Andaman Sea (B). PMID- 15826496 TI - A modified mutation detection method for large-scale cloning of the possible single nucleotide polymorphism sequences. AB - Although the human genome has been nearly completely sequenced, the functions and the roles of the vast majority of the genes, and the influences of single nucleotide polymorphisms (SNPs) in these genes are not entirely known. A modified mutation detection method was developed for large-scale cloning of the possible SNPs between tumor and normal cells for facilitating the identification of genetic factors that associated with cancer formation and progression. The method involves hybridization of restriction enzyme-cut chromosomal DNA, cleavage and modification of the sites of differences by enzymes, and differential cloning of sequence variations with a designed vector. Experimental validations of the presence and location of sequence variations in the isolated clones by PCR and DNA sequencing support the capability of this method in identifying sequence differences between tumor cells and normal cells. PMID- 15826497 TI - Interaction of resveratrol and genistein with nucleic acids. AB - Resveratrol (RES) and genistein (GEN) are the dietary natural products known to possess chemopreventive property and also the ability to repair DNA damage induced by mutagens/carcinogens. It is believed that the therapeutic activity of these compounds could be primarily due to their interaction with nucleic acids but detailed reports are not available. We here explore the interaction of these drugs with nucleic acids considering DNA and RNA as a potential therapeutic target. The interaction of RES and GEN has been analysed in buffered solution with DNA [saline sodium citrate (SSC)] and RNA [tris ethylene diammine tetra acetic acid (TE)] using UV-absorption and Fourier transform infrared (FTIR) spectroscopy. The UV analysis revealed lesser binding affinity with nucleic acids at lower concentration of RES (P/D = 5.00 and 10.00), while at higher drug concentration (P/D = 0.75, 1.00 and 2.50) hyperchromic effect with shift in the lambda(max) is noted for DNA and RNA. A major RES-nucleic acids complexes was observed through base pairs and phosphate backbone groups with K = 35.782 M(-1) and K = 34.25 M(-1) for DNARES and RNA-RES complexes respectively. At various concentrations of GEN (P/D = 0.25, 0.50, 0.75, 1.00 and 2.50) hyperchromicity with shift in the lambda(max) from 260-->263 nm and 260--> 270 nm is observed for DNA-GEN and RNA-GEN complexes respectively. The binding constant (from UV analysis) for GEN-nucleic acids complexes could not be obtained due to GEN absorbance overlap with that of nucleic acids at 260 nm. Nevertheless a detailed analysis with regard to the interaction of these drugs (RES/GEN) with DNA and RNA could feasibly be understood by FTIR. PMID- 15826498 TI - Cloning and expression analysis of gonadogenesis-associated gene SPATA4 from rainbow trout (Oncorhynchus mykiss). AB - Gonadogenesis is a complicated process which involves multi-gene interactions. A rainbow trout (Oncorhynchus mykiss) gene spermatogenesis associated 4 (SPATA4) was cloned and characterized from adult rainbow trout testis. The cDNA sequence of rainbow trout SPATA4 contains an open reading frame of 1, 081 nucleatides encoding a putative protein of 259 amino acids. The putative protein from rainbow trout shares a 76.8% homology with zebrafish SPATA4. No trans-membrane regions or signal peptide were detected using bioinformatics methods. Subcellular localization analysis revealed that rainbow trout SPATA4 was a nuclear protein with highest possibility (39.1%). Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the distribution of rainbow trout SPATA4 in eleven organs of adult rainbow trout. The result demonstrated that this gene express specifically in testis and slight amount of expression was detected in ovary. Further analysis of SPATA4 characterization and function in rainbow trout may provide insight into the understanding of gonadogenesis process. PMID- 15826499 TI - 4-acetoxyscirpendiol of Paecilomyces tenuipes inhibits Na(+)/D-glucose cotransporter expressed in Xenopus laevis oocytes. AB - Cordyceps, an entomopathogenic fungus, contains many health-promoting ingredients. Recent reports indicate that the consumption of cordyceps helps reduce blood-sugar content in diabetics. However, the mechanism underlying this reduction in circulatory sugar content is not fully understood. Methanolic extracts were prepared from the fruiting bodies of Paecilomyces tenuipes, and 4 beta acetoxyscirpendiol (4-ASD) was eventually isolated and purified. Na(+)/Glucose transporter-1 (SGLT-1) was expressed in Xenopus oocytes, and the effect of 4-ASD on SGLT-1 was analyzed utilizing a voltage clamp and by performing 2-deoxy-D-glucose (2-DOG) uptake studies. 4-ASD was shown to significantly inhibit SGLT-1 activity compared to the non-treated control in a dose-dependent manner. In the presence of the derivatives of 4-ASD (diacetoxyscirpenol or 15-acetoxyscirpendiol), SGLT-1 activity was greatly inhibited in an 4-ASD-like manner. Of these derivatives, 15-acetoxyscirepenol inhibited SGLT-1 as well as 4-ASD, whereas diacetoxyscirpenol was slightly less effective. Taken together, these results strongly indicate that 4-ASD in P. tenuipes may lower blood sugar levels in the circulatory system. We conclude that 4-ASD and its derivatives are effective SGLT-1 inhibitors. PMID- 15826500 TI - Enhanced antioxidant enzymes are associated with reduced hydrogen peroxide in barley roots under saline stress. AB - Antioxidant enzymes are related to the resistance to various abiotic stresses including salinity. Barley is relatively tolerant to saline stress among crop plants, but little information is available on barley antioxidant enzymes under salinity stress. We investigated temporal and spatial responses of activities and isoform profiles of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), non-specific peroxidase (POX), and glutathione reductase (GR) to saline stress in barley seedlings treated with 200 mM NaCl for 0, 1, 2, 5 days, respectively. In the control plant, hydrogen peroxide content was about 2 fold higher in the root than in the shoot. Under saline stress, hydrogen peroxide content was decreased drastically by 70% at 2 d after NaCl treatment (DAT) in the root. In the leaf, however, the content was remained unchanged by 2 DAT and increased about 14 % at 5 DAT. In general, the activities of antioxidant enzymes were increased in the root and shoot under saline stress. But the increase was more significant and consistent in the root. The activities of SOD, CAT, APX, POX, and GR were increased significantly in the root within 1 DAT, and various elevated levels were maintained by 5 DAT. Among the antioxidant enzymes, CAT activity was increased the most drastically. The significant increase in the activities of SOD, CAT, APX, POX, and GR in the NaCl-stressed barley root was highly correlated with the increased expression of the constitutive isoforms as well as the induced ones. The hydrogen peroxide content in the root. PMID- 15826501 TI - Expression of porcine acid-labile subunit (pALS) of the 150-kilodalton ternary insulin-like growth factor complex and initial characterization of recombinant pALS protein. AB - Acid-labile subunit (ALS) is a component of the 150-kDa insulin-like growth factor-binding protein-3 (IGFBP-3) complex, which, by sequestering the majority of IGFs-I and -II and thereby prolonging the half-life of them in plasma, serves as a circulating reservoir of IGFs in mammalian species. A pGEX-2T plasmid and a baculovirus expression constructs harboring a coding sequence for glutathione-S transferase (GST)-porcine ALS (pALS) fusion protein were expressed in BL21(DE3) E. coli and Sf9 insect cells, respectively. The expressed protein was purified by glutathione or Ni-NTN affinity chromatography, followed by cleavage of the fusion protein using Factor Xa. In addition, pALS and hIGFBP-3 were also produced in small amounts in the Xenopus oocyte expression system which does not require any purification procedure. A 65-kDa pALS polypeptide was obtained following the prokaryotic expression and the enzymatic digestion, but biochemical characterization of this polypeptide was precluded because of an extremely low expression efficiency. The baculovirus as well as Xenopus-expressed pALS exhibited the expected molecular mass of 85 kDa which was reduced into 75 and 65 kDa following deglycosylation of Asn-linked carbohydrates by Endo-F glycosidase, indicating that the expressed pALS was properly glycosylated. Moreover, irrespective of the source of pALS, the recombinant pALS and hIGFBP-3 formed a 130-kDa binary complex which could be immunoprecipitated by anti-hIGFBP-3 antibodies. Collectively, results indicate that an authentic pALS protein can be produced by the current expression systems. PMID- 15826502 TI - Purification and biochemical properties of glutathione S-transferase from Lactuca sativa. AB - A glutathione S-transferase (GST) from Lactuca sativa was purified to electrophoretic homogeneity approximately 403-fold with a 9.6% activity yield by DEAE-Sephacel and glutathione (GSH)-Sepharose column chromatography. The molecular weight of the enzyme was determined to be approximately 23,000 by SDS polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the enzyme was significantly inhibited by ShexylGSH and S-(2,4-dinitrophenyl) glutathione. The enzyme displayed activity towards 1-chloro-2,4-dinitrobenzene, a general GST substrate and high activities towards ethacrynic acid. It also exhibited glutathione peroxidase activity toward cumene hydroperoxide. PMID- 15826503 TI - The association between the T102C polymorphism of the HTR2A serotonin receptor gene and HDL cholesterol level in Koreans. AB - 5-HT2A is one of major serotonin receptor that is involved in the action of serotonin-targeting drugs. Previous clinical studies have shown an unexpected association between lower cholesterol level and psychiatric diseases, in which T102C polymorphism of HTR2A, gene of 5-HT2A serotonin receptor, might be involved. Therefore, we hypothesized a potential association between lower cholesterol level and T102C polymorphism. The effect of the T102C polymorphism on the serum lipid profiles of 646 subjects without specific psychiatric disease was investigated. Genotype was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. There were significantly lower levels of total cholesterol (193.6 +/- 35.0 versus 202.1 +/- 45.5 mg/dl, p = 0.016) and HDL-cholesterol (42.7 +/- 11.6 versus 46.3 +/- 12.7 mg/dl, p = 0.004) in CC genotype than non-CC genotypes. Moreover, multivariate analysis showed that the CC genotype is a strong predictor of a lower HDL-cholesterol level (p < 0.001). In conclusion, this study shows that the CC genotype of the HTR2A gene is related to lower HDL-cholesterol level in Koreans. This is the first demonstration showing the potential genetic relationship between the serotonin receptor gene polymorphism and the HDL-cholesterol level. PMID- 15826504 TI - Secondary structure, 1H, 13C and 15N resonance assignments and molecular interactions of the dishevelled DIX domain. AB - Dishevelled (Dvl) is a positive regulator of the canonical Wnt signaling pathway, which regulates the levels of beta-catenin. The beta-catenin oncoprotein depends upon the association of Dvl and Axin proteins through their DIX domains, and its accumulation directs the expression of specific developmental-related genes at the nucleus. Here, the (1)H, (13)C and (15)N resonances of the human Dishevelled 2 DIX domain are assigned using heteronuclear nuclear magnetic resonance (NMR) spectroscopy. In addition, helical and extended elements are identified based on the NMR data. The results establish a structural context for characterizing the actin and phospholipid interactions and binding sites of this novel domain, and provide insights into its role in protein localization to stress fibers and cytoplasmic vesicles during Wnt signaling. PMID- 15826505 TI - Asparagine-473 residue is important to the efficient function of human dihydrolipoamide dehydrogenase. AB - Dihydrolipoamide dehydrogenase (E3) catalyzes the reoxidation of dihydrolipoyl moiety of the acyltransferase components of three alpha-keto acid dehydrogenase complexes and of the hydrogen-carrier protein of the glycine cleavage system. His 457 of Pseudomonas putida E3 is suggested to interact with the hydroxyl group of Tyr-18 of the other subunit and with Glu-446, a component in the last helical structure. To examine the importance of the suggested interactions in human E3 function, the corresponding residue of human E3, Asn-473, was substituted to Leu using site-directed mutagenesis. The E3 mutant was expressed in Escherichia coli and highly purified using an affinity column. Its E3 activity was decreased about 37-fold, indicating that Asn-473 residue was important to the efficient catalytic function of human E3. Its slightly altered spectroscopic properties implied that small conformational changes could occur in the E3 mutant. PMID- 15826506 TI - Glycoproteins contained within Soamsan, a traditional Oriental medicine, are the main class of active ingredients responsible for the medicine-induced immune stimulation. AB - In our previous study, Soamsan, a traditional Oriental medicine, was shown to enhance the induction of antigen-specific immune responses, and it was speculated that the enhancing activity might be closely associated with glycoproteins contained within the medicine. To elucidate this speculation, protein samples from each component, used in the preparation of Soamsan, were obtained and their immune stimulating activities were tested with mouse splenocytes. All the samples markedly enhanced the lymphocyte proliferation and cytokine secretion by the mouse splenocytes. In particular, the enhancement was significantly higher with the protein sample treatments than with those of the original crude sample. Furthermore, the pronase E- and NaIO4-mediated inhibition of splenocyte stimulation activity of the protein samples clearly supported that glycoproteins are the main class of active ingredients responsible for the lymphocyte stimulating activity of the samples. Consequently, our findings suggest that glycoproteins might have a pivotal role in Soamsan-mediated immune modulation, although the in vivo effect of the glycoproteins should be further elucidated. PMID- 15826507 TI - [The clinical relationship today]. PMID- 15826508 TI - [Canada: family medicine. Crisis and response]. PMID- 15826509 TI - [Improving communication between levels of health care: direct referral of patients to a one-stop service for major outpatient surgery]. AB - OBJECTIVE: To analyze the results obtained with a "one-stop" specialty service designed as part of a collaborative program involving primary and specialized care in order to improve communication between levels of care and reduce the delay in referral and surgical treatment for patients eligible for outpatient surgery. DESIGN: Prospective, descriptive, longitudinal study. SETTING: Major Outpatient Surgery Unit of the Hospital Clinico San Carlos and health centers serving Health Area 7 in Madrid, central Spain. PARTICIPANTS: Patients more than 14 years of age with a surgical condition involving the abdominal wall, pilonidal sinus, soft-tissue tumor, or proctological disease. INTERVENTION: Direct referral, with completed preoperative work-up, of patients from health centers to the Major Outpatient Surgery Unit of the Hospital Clinico San Carlos according to a protocol developed by consensus. The patient is seen on the same day for surgical work-up and anesthesia work-up, and is given preoperative information. Patients then make only one further visit to the hospital to undergo surgery. MAIN MEASURES: Number of patients with each diagnosis referred, diagnostic concordance between the health center and hospital, delay from referral to surgical treatment, number of trips made for different appointments, and referral rate. RESULTS: A total of 188 patients were referred. More than two thirds (68.7%) had an abdominal wall condition eligible for direct referral. Diagnostic concordance was 96%. The delay from referral until surgery was reduced by 60%, and the number of trips for appointments was reduced by 66.6%. The overall referral rate was 12.6%. CONCLUSIONS: Because of its feasibility, acceptability, and cost-efficiency, the direct referral system has the potential to improve relations between primary and specialized care and enhance the quality of care by shortening the delay to treatment. PMID- 15826510 TI - [Can communication between levels of care be improved?]. PMID- 15826511 TI - [The opinion of the users as opportunity for improvement in primary health care]. AB - OBJECTIVES: To determine those aspects of Primary Care Center (PCC) that displeases more the clients/users, to investigate its determinants, and to apply the intervention or corresponding interventions. DESIGN: Results evaluation oriented. Detection of dissatisfaction reasons through cross observational study from survey self-fulfilled on 200 individuals selected through random cluster sampling. A continuous improvement group was implemented in order to the factors causing detected problems and their possible solutions were analyzed. LOCATION: Rambla PCC, Sant Feliu de Llobregat. Catalonian Health Institute. PARTICIPANTS AND CONTEXT: Users/clients of the PCC who were in the waiting room. Improvement Group: Professionals of Sant Feliu Primary Care Team and Baix Llobregat Nord Primary Care Service. INTERVENTIONS: Reorganization of the family doctors' agendas and reduction of bureaucratic tasks in the physicians' offices. PRINCIPAL MEASUREMENTS: Respondents socio-demographic data, and reasons for their dissatisfaction. Determinants of the problem were analyzed. The principal cause of discomfort was the delay of previous appointment. RESULTS: Among reasons for discomfort expressed by the users, the most frequent was lag time with 16% (95% CI, 10.9-21.1). This complaint referred both to wait for obtaining visit and to be in the waiting room before being visited. As determinants of delay there were identified a high number of patients, visits duplicity, bureaucratic visits, high number of urgent visits and an insufficient previous appointment visits. CONCLUSIONS: Patient's opinions help us to know how public health services are accomplishing their expectations. Process improvement is indispensable in order to provide a quality product, but it must be endowed with resources adequacy to population needs. PMID- 15826512 TI - [Breast-feeding: can health staff positively affect its duration?]. AB - INTRODUCTION: Mother's milk is the ideal meal for the baby during the first six months of life. A good health education (before and after birth) helps to prolong breastfeeding (BF). OBJECTIVE: To know the prevalence of BF among a group of women and to study the relationship with health education. DESIGN: Observational and analytic study. SETTING: Dr. Pujol i Capsada Primary Health Center (El Prat de Llobregat); Casagemes CAD's afterbirth group in Badalona, and Alba Lactancia Women's group. SUBJECTS OF THE STUDY: 135 women with babies born between the 1st of March 2002 and the 28th of February 2003. METHOD: Mother's motivation for breast-feeding and the type of information she has got were evaluated. Breast feeding period was measured from the second month of baby's life by means of a survey passed by nursing staff. RESULTS: The average BF period was of 160 days, 22% breastfed their babies for less than a week, and 14% between a week and a month. The main variables in relation with the increase of BF are: to want to breast-feed (P=.05); mother sleeping with the baby (P=.03); to receive assistance from nursing staff while in hospital (P=.01); not to receive additional meals in hospital (P=.02); no problems during the first month of life (P<.0001); to receive information from health center (P=.009). If the first contact mother-baby (bringing the baby close to the breast) lasts more than 30 minutes 51% give up BF before the first month; but if the first contact is before of 30 minutes only give up 20.8% (P=.002). CONCLUSIONS: The prevalence of BF increases among the women that got health education and support from the health professionals. PMID- 15826513 TI - [Professional burnout in dentists and stomatologists of the Galician Health Service]. AB - AIM: To analyse the degree of professional burnout in primary care dentists of the Galician Health Service (GHS). DESIGN: Descriptive, cross-sectional study. SETTING: Dental care units of the GHS primary care network. SUBJECTS: All primary care dentists of the new primary care model of the GHS (N=79). Response rate: 50% (N=35). MEASUREMENTS: Anonymous self-applied questionnaire: the Maslach Burnout Inventory, that evaluates emotional exhaustion, depersonalization, and personal achievements; a social and personal questionnaire. Student's t test, ANOVA, 2 or Fisher's exact test were used. MAIN RESULTS: Mean age: 46.58+/-9.47; sex: 74.3% males; temporary job: 82.9%; years in the GHS: 15.25+/-8.34. High values were detected in: emotional exhaustion 54.3%; depersonalization 55.6%; personal achievements 6.9%. No statistically significant differences were identified in terms of age, sex, marital status, type of contract, rural vs urban areas, number of patients treated, or years in the GHS, apart from the "personal achievements" subscale, that was rated, significantly higher (P=.046) by those who had shorter experience at for the GHS. CONCLUSION: GHS dentists show a degree of professional burnout close to the one described for family doctors at the Spanish National Health System. PMID- 15826514 TI - [Emotion and intuition as tools to deal with uncertainty when taking decisions in family practice]. PMID- 15826515 TI - [Complementary and alternative medicine: some reflections on bioethics]. PMID- 15826516 TI - [Impact of vaccination programmes in Spain]. PMID- 15826517 TI - [How to organise teaching consultations for training residents. Some basic principles]. PMID- 15826518 TI - [Probable cardiac failure due to rofecoxib]. PMID- 15826519 TI - [Sepsis due to PSA]. PMID- 15826520 TI - [Home care patients: a day in their life]. PMID- 15826521 TI - [Prevalence of thyroid dysfunction among the elderly]. PMID- 15826523 TI - [Few clinical trials, and these with poorly reported results]. PMID- 15826524 TI - [Serious haemorrhages linked to acenocoumarol in a rural area]. PMID- 15826525 TI - [Changes in the surgical treatment of bronchogenic cancer]. PMID- 15826526 TI - [Non-small cell bronchogenic cancer in stage IA: mortality patterns after surgery]. AB - OBJECTIVE: To determine the causes of death in patients treated surgically for nonsmall cell lung cancer (NSCLC) in stage IA and to evaluate the impact on survival of not performing systematic lymph node dissection and of the number of nodes resected. PATIENTS AND METHODS: The study sample consisted of 156 patients operated on for NSCLC and classified in stage IA according to TNM staging. Only palpable or visible lymph nodes were dissected. Kaplan-Meier survival curves were compared using a log-rank test. RESULTS: At the end of the study, 85 (54.5%) patients had died, 67 (42.9%) were alive, and 4 (2.5%) were lost to follow up. Twenty-three (14.7%) died from a recurrence of NSCLC: 2 with local tumors (1.2%), 2 with mediastinal node involvement (1.2%), and 19 (12.1%) with distant metastasis. The cause of death was unrelated to NSCLC in 62 (39.7%) cases: 33 (21.1%) had a new tumor, 18 of which were bronchogenic, and 29 (18.5%) had nonmalignant disease. The 5-year survival rate was 81.4%. The rate was 88.9% among patients from whom no lymph nodes were excised and 79.9% among those with node excision, although the difference was not statistically significant (P=.4073). CONCLUSIONS: Our experience suggests that neither the fact of not performing systematic lymph node dissection nor the number of nodes resected has an impact on survival. A substantial number of patients died of causes unrelated to the NSCLC for which they had been treated. PMID- 15826527 TI - [Prognostic value of the carcinoembryonic antigen found in pleural lavage fluid from patients with lung carcinoma]. AB - OBJECTIVE: To detect the tumor marker carcinoembryonic antigen (CEA) in pleural lavage fluid taken during surgery from patients with pulmonary carcinoma without associated pleural effusion and assess its possible prognostic implications. PATIENTS AND METHODS: A prospective, observational study was undertaken to include consecutive patients who underwent surgical treatment for lung carcinoma in which pleural lavage was performed prior to closure of the thoracic cavity (study group). The same techniques and measurements were used in patients undergoing thoracotomy for benign disease (control group). The preoperative blood level of CEA was also quantified. RESULTS: In the study group, the median CEA levels in blood and pleural lavage fluid were 2.90 ng/mL and 0.40 ng/mL respectively; these figures are higher than those corresponding to the control group. A CEA level of 0.30 ng/mL in pleural lavage fluid was established as a cutoff point, based on the corresponding receiver operating characteristic curve, with a sensitivity of 68.4% and a specificity of 35.7%. A graph of survival in relation to this cutoff point revealed a statistically significant effect (P<.05). CONCLUSIONS: It is possible to detect CEA in pleural lavage fluid from the thoracic cavity of patients with lung carcinoma. The values obtained are higher than those found in fluid from patients without neoplastic disease, and this parameter functions as an independent predictor of disease course. PMID- 15826528 TI - [Clinical and epidemiological study of disease caused by Mycobacterium kansasii in the metropolitan area of Bilbao, Spain]. AB - OBJECTIVE: Epidemiological description of individuals from whom Mycobacterium kansasii isolates were obtained in respiratory samples, and analysis of the isolates using molecular biological techniques. MATERIAL AND METHODS: A descriptive retrospective/ prospective study was carried out from January 1994 to April 2002 in Basurto Hospital and Santa Marina Hospital and from January 2000 to April 2002 in Cruces Hospital, Galdakao Hospital, and San Eloy Hospital. Diagnosis of the disease was performed according to American Thoracic Society criteria; other definitions were also applied to allow inclusion of all cases. Disease caused by M. kansasii in patients who were not infected with the human immunodeficiency virus (HIV) was compared with disease caused by Mycobacterium tuberculosis in a control group. Polymerase chain reaction was applied with analysis of restriction fragment length polymorphisms to differentiate between species of mycobacteria and classify them into genotypes. Amplified fragment length polymorphisms were used to recognize clones within each genotype. RESULTS: The patient charts of 334 patients in which an isolate of M. kansasii had been recorded were reviewed. We considered 220 patients to be suffering from disease caused by M. kansasii (American Thoracic Society criteria along with probable disease according to established definitions). The disease was more frequent in male patients (n=185; 84.1%) and in individuals who were not HIV positive (n=184; 83.6%). The highest incidence of disease in the Bizkaia region was found in Margen Izquierda-Encartaciones, where the rate was 8.05 per 100 000 inhabitants. In the Bilbao area, the highest rate was found in the districts lying on the outskirts. The underlying diseases were tuberculosis (20.5%), chronic obstructive pulmonary disease (25.9%), pulmonary neoplasia (7.7%), silicosis (0.9%), chronic liver disease (11.4%), and duodenal ulcer (8.6%). The most frequent constitutional symptoms were fever (39.1%), loss of appetite (23.2%), and weight loss (33.3%). Among the respiratory symptoms, the most outstanding were cough (70.9%) and expectoration (62.3%). The most frequent radiographic patterns were cavitation and pulmonary infiltration. The most common treatment regimen was rifampicin, isoniazid, and ethambutol (43.4%), and the average duration was 12 months in patients who were HIV negative. Analysis of antibiotic sensitivity, performed on 56 strains, revealed that 100% were resistant to isoniazid, while none displayed rifampicin resistance. Thirty-four cases of disease caused by M. kansasii were compared with 68 cases of tuberculosis, all of them without HIV infection. The comparison revealed a predominance of smokers, respiratory symptoms, and cavitation in patients with disease caused by M. kansasii. The majority of the isolates (98.5%) corresponded to genotype I. A total of 8 clones were obtained; the clones designated 1 and 3 were more common in HIV-positive and HIV-negative individuals respectively. CONCLUSIONS: In recent years, there has been an increase in the number of patients with disease caused by M. kansasii in the province of Bizkaia. The disease is more frequent in male patients, individuals who are HIV negative, and in urban areas. In addition, more respiratory symptoms and a higher incidence of cavitation were found in patients with disease caused by M. kansasii than in those with tuberculosis. Genotype I is the most common isolate, and clones 1 and 3 affect 80% of patients suffering from the disease. PMID- 15826529 TI - [Results of a comprehensive workplace program for the prevention and treatment of smoking addiction]. AB - OBJECTIVES: To assess a comprehensive smoking prevention and treatment program in an electrical appliances company with 1600 employees. MATERIAL AND METHODS: The program included smoking restrictions with the designation of smoking areas and the offer of smoking cessation treatment for the smokers affected. Study variables were age, sex, nicotine dependence (Fagerstrom Test), carbon monoxide in expired air, adherence to therapy, and smoking abstinence at 1 week, 1 month, 3 months (end of treatment), and 6 months. Successful smoking abstinence was defined as continuous abstinence from the beginning of treatment. RESULTS: Smoking prevalence was 34.8% and 19.5% of smokers requested treatment (77.4% men and 22.6% women). Mean (SD) age was 41.3 (10.3) years. Mean score of nicotine dependence was 5.3 (2.6) and the mean quantity of carbon monoxide in expired air was 35.6 (23.7) ppm. Adherence to therapy was good in 80% of patients. Rate of abstinence was 57.5% at 6 months, signifying a 4% reduction in prevalence. CONCLUSIONS: Workplace smoking cessation programs reduce prevalence and facilitate the establishment of smoking restrictions at the worksite. Companies are convenient settings for the implementation of programs aimed at smoking prevention and treatment. PMID- 15826530 TI - [Lung cancer risk in shoe manufacturing]. AB - OBJECTIVE: Workers in shoe manufacturing have been reported to be at a greater relative risk for bronchogenic carcinoma. Given the implications for our practice setting, we carried out a study to a) clarify whether working in shoe manufacturing is a risk factor for lung cancer and b) detect histological differences between lung cancers in shoe manufacturers and in other lung cancer patients. PATIENTS AND METHODS: This case-control study compared all lung cancer patients diagnosed in Hospital Elda, Alicante, Spain, between January 1994 and December 1999, with a control group composed of patients admitted to the same hospital for accidental fractures. Information on occupational history and tobacco dependency was collected from all patients by telephone questionnaire. RESULTS: One hundred and ninety-one case patients and 192 control patients were included in the study; 52 of the cases (27.2%) and 48 controls (25%) worked in shoe manufacturing. No statistically significant differences were found between the 2 groups, not even when we limited the cases and controls to only those who had worked more than 30 years in shoe manufacturing or when we analyzed only subjects who had had especially high risk occupations. No differences in tumor histology were found between cancer patients who worked in shoe manufacture and those who did not. CONCLUSIONS: Working in shoe manufacturing has not proven to be a risk factor for bronchogenic carcinoma. PMID- 15826531 TI - [Respiratory infections caused by environmental mycobacteria]. PMID- 15826532 TI - [Hospital treatment of chronic obstructive pulmonary disease exacerbations: an evidence-based review]. PMID- 15826533 TI - [Bronchiectasis following colectomy in a patient with ulcerative colitis and factor V Leiden mutation]. AB - For some years it has been known that lung disease may be present in patients with inflammatory bowel disease (IBD). Diseases of the central airway, bronchi, and lung parenchyma are among the most common forms of lung involvement in IBD patients. Bronchiectasis is frequent and almost always appears after the onset of disease and in close association with inflammatory activity. However, reports of the appearance of bronchiectasis following colectomy for an exacerbation of IBD have been rare. We present the case of a 36-year-old man with severe ulcerative colitis who, following a total colectomy, developed bilateral bronchiectasis 12 months after surgery and for whom both preoperative and postoperative computed tomography scans were available. PMID- 15826534 TI - [Bronchioloalveolar adenoma associated with bronchiolitis obliterans and leishmaniasis with lung involvement in acquired immunodeficiency syndrome]. AB - Visceral leishmaniasis is not unusual in patients with acquired immunodeficiency syndrome (AIDS), but lung infiltration is uncommon. Leishmaniasis involving the lung often manifests as interstitial pneumonitis. We report a case in which the discovery of amastigotes in the transbronchial biopsy led to a diagnosis of leishmaniasis. However, the findings from x-rays and study of the bronchoalveolar lavage fluid were consistent with bronchiolitis obliterans, possibly caused by the AIDS virus. In addition, the transbronchial biopsy findings were consistent with a diagnosis of bronchioloalveolar adenoma with radiographic evidence of multiple nodules. PMID- 15826535 TI - [Intraosseous lipoma of rib]. PMID- 15826536 TI - [Hospitalization costs for chronic obstructive pulmonary disease in Andalusia, Spain: 1998 through 2002]. PMID- 15826538 TI - [Algorithms are always useful for the diagnosis of pulmonary embolism]. PMID- 15826539 TI - [Impact of a hospital antibiotic policy]. PMID- 15826540 TI - [Impact of a non-compulsory antibiotic control program (PACTA): cost reductions and decreases in some nosocomial infections]. AB - BACKGROUND: Antibiotics account for 30% of hospital pharmacy expenses. More than 50% of the prescriptions are considered inappropriate; hence, programs devoted to optimizing the prescription of antibiotics should be developed. We present the results of a non-compulsory program for the assessment and control of antibiotic treatment in the University Hospital 12 de Octubre in Madrid. METHODS: The program was applied in the hospitalization units of six medical and surgical departments. Treatments in all patients were checked daily and recommendations were made in writing, according to previously established criteria. The program was used for 12 months and the results were compared with those of the previous 12 months. RESULTS: 1,280 treatments were reviewed and 524 recommendations were made (80% of them were accepted). There was a 13.82% reduction in the number of defined daily doses of antibiotics/100 inpatient-days. Antibiotic expenditure decreased by 65,352 euros (5,446 euros/month), implying a reduction of 1.21 euros/hospitalization-bed/day. There were no statistically significant differences in length of hospital stay or mortality between the two periods. A reduction in the incidence of Clostridium difficile diarrhea (p < 0.0001) and Candida spp. isolations (p < 0.05) was observed. CONCLUSIONS: Following application of a non-compulsory control program, antibiotic prescription improved and expenditure decreased, with no change in length of hospital stay or mortality. There was a reduction in the incidence of some nosocomial infections. Acceptation of the program by the physicians of the departments implicated was favorable. PMID- 15826541 TI - [Dermatophytosis with concurrent lesions in distant locations. Prognostic and therapeutic significance]. AB - INTRODUCTION: Recurrence and a lack of treatment response are common in dermatophytosis. In patients with cutaneous and concurrent toenail lesions, often only the former are investigated, which may result in inappropriate treatment due to misdiagnosis. METHODS: Between January 2000 and April 2004, we prospectively studied the presence of dermatophytic fungi in lesions other than those prompting the consultation to further investigate the diagnosis. RESULTS: We prospectively identified 61 patients with dermatophytosis and concurrent lesions caused by the same fungus at sites distant from the primary lesion. Concurrent lesions occurred in 15.9% of culture-confirmed dermatophyte infections. Thirty-six patients (59%) consulted for skin lesions located at sites other than that of the primary lesion in the foot. The most frequently identified species was Trichophyton rubrum (50/61, 82%). CONCLUSIONS: Localization of all the lesions, as well as isolation and identification of the causative fungus, are essential to establish the prognosis and choose the most appropriate antifungal agent, route of administration, and duration of treatment in dermatophytosis. PMID- 15826542 TI - [Clinical and epidemiological study of spotted fever group rickettsiosis in Albacete, Spain]. AB - INTRODUCTION: The aim of this study was to describe the clinical and epidemiological features of spotted fever group rickettsiosis acquired in the province of Albacete, Spain. METHODS: We retrospectively reviewed the clinical records of patients with a diagnosis of spotted fever group rickettsiosis and positive serologic results between 1997 and 2003. Criteria for inclusion were as follows: 1) seroconversion in IgG to Rickettsia conorii by indirect immunofluorescence, or 2) a single titer of 160 dollars with two or three of the following symptoms: fever, eschar, or rash. RESULTS: There were 41 cases in the study period. Mean age was 58 years (range: 14-84). Thirty-seven (90%) patients were 40 or more years old (RR: 10,1; 95% CI: 4,3-24,0). Twenty-five (63%) patients were of rural origin (RR: 3,5; 95% CI: 1,8-6,5). Thirty-six (88%) patients had regular or recent contact with the rural milieu or with animals. Forty patients had fever and 38 (93%) had one or two eschars. There were 32 cases of Mediterranean spotted fever, 2 cases with symptoms consistent with DEBONEL/TIBOLA, and 7 atypical cases with eschar and no rash. CONCLUSION: Spotted fever group rickettsiosis was mainly found in adults in contact with the rural milieu or with animals. Age over 40 years old and residence in a rural area were risk factors. Mediterranean spotted fever was the most common syndrome. PMID- 15826543 TI - [Changes in the antimicrobial susceptibility of Escherichia coli isolates from nosocomial versus community-acquired urinary tract infections]. AB - INTRODUCTION: We studied the variations in Escherichia coli sensitivity patterns to commonly used antimicrobial agents in urinary tract infections, by stratifying isolates according to year and source. MATERIAL AND METHODS: Retrospective analysis of the sensitivity of 14,319 E. coli urine isolates to 14 antimicrobials during the period of 1994-2001. Sensitivity comparison by source of the isolate began in 1996, and included 13,263 isolates originating in the hospital and 2,350 originating in the community. RESULTS: Penicillins were the least active antibiotic agents against the E. coli isolated (sensitivity 40.9%), followed by cotrimoxazole (66.4%), nalidixic acid (70.8%) and norfloxacin (76.1%). Sensitivities to the other antimicrobials tested remained near 95%. Sensitivity of E. coli isolates to most of the antimicrobials tested decreased gradually during the 8-year period, with a marked decrease for nalidixic acid and norfloxacin (76.0% down to 63.3% and 85.1% down to 66.6%, respectively). Nevertheless, sensitivity of E. coli to nitrofurantoin and ampicillin sulbactam/amoxicillin-clavulanic acid increased; no significant differences were observed regarding cefazolin, gentamicin and fosfomycin. Sensitivity was higher in community E. coli isolates than in nosocomial isolates. The greatest differences corresponded to cephalosporins (OR = 2), nitrofurantoin (OR = 1.72), and ampicillin-sulbactam/ amoxicillin-clavulanic acid (OR = 1.57). Antimicrobials with significant differences in sensitivity between isolates of different sources decreased gradually from 6 (1996) to 1 (2001). CONCLUSIONS: Penicillins, cotrimoxazole and quinolones can no longer be considered the antimicrobials of choice for empirical treatment of E. coli urinary tract infections. Along the study period, we observed a reduction in the initial susceptibility differences among hospital and community isolates. PMID- 15826544 TI - [Factors predicting the failure of interruption of highly active antiretroviral treatment (HAART) in HIV-infected patients]. AB - INTRODUCTION: Treatment interruptions may be an alternative to HAART in the management of chronically infected HIV-patients. We designed this study in an attempt to assess the predictability of this strategy. METHODS: We recruited HIV infected patients whose treatment had been suspended. Interruption was due to the patient's own decision, or toxicity, or because the patient had started the treatment with more than 350 CD41 cells/microL (immunologic criteria). RESULTS: Forty-one consecutive patients were included, with a median follow-up of 13 months. Failure was associated with the reason for interruption (p 5 0.0063). Failure occurred in 14.3% of those who interrupted treatment due to immunological criteria and in 40% of those who interrupted treatment due to their own decision or toxicity. The reasons for interruption were: toxicity in 11 patients (26.8%), personal decision in 9 (21.9%) and immunological criteria in 21 (51.2%). In the univariate analysis, the nadir CD41 cell count < 350 cel./microL [OR 16 (p = 0.054)] was statistically significant in the patients who stopped treatment due to immunological criteria, while treatment with protease inhibitors [OR 14 (p = 0.032)] was statistically significant in the remaining patients. In the multivariable analysis only nadir CD41 < 350 cel./microL was independently related with failure. CONCLUSIONS: Failure was related to interruption criteria and was greater in patients who stopped due their own decision or toxicity. When interruption was due to immunological criteria, the factor predicting failure was nadir CD41 cell count < 350 cel./microL. In the remaining patients, none of the variables was related to failure. PMID- 15826545 TI - [Detection of methicillin resistance and identification of Staphylococcus spp. from positive blood culture bottles using the mecA and nucA genes with the LightCycler System]. AB - INTRODUCTION: The aim of this study was to evaluate the feasibility of detecting Staphylococcus aureus and coagulase-negative staphylococci (CoNS) and of identifying methicillin resistance directly in positive BACTEC blood culture bottles using the LightCycler system. METHODS: One hundred thirty-one positive blood culture bottles in which Gram-positive cocci in cluster were observed after Gram staining and 40 positive bottles with microorganisms other than staphylococci were studied. A molecular assay based on an automated DNA extraction protocol with a MagNA Pure LC instrument was used. Oligonucleotide primers and fluorescence-labeled hybridization probes were designed for amplification and sequence-specific detection of both a 408-pb fragment within the mecA gene and a 279-pb fragment within the S. aureus-specific nucA gene. RESULTS: All the bottles that yielded methicillin-resistant S. aureus (MRSA), methicillin-sensitive S. aureus (MSSA) or methicillin-resistant CoNS (MRCoNS) strains were correctly identified by the nucA and mecA PCR assays. One bottle that yielded a mixed culture of MSSA and MRCoNS gave positive results for both genes. In the 21 bottles with methicillin-susceptible CoNS (MSCoNS), nucA PCR were negative, but two of these bottles gave positive results for the mecA gene. The sensitivity and specificity of the nucA gene assay were 100%. The sensitivity and specificity of the PCR assay for detection of methicillin resistance with the mecA gene were 100% and 97.5%, respectively. CONCLUSION: This is a sensitive and highly specific method for identifying staphylococci in positive blood cultures, allowing discrimination between methicillin-susceptible and -resistant strains in less than 3 hours after Gram stain. PMID- 15826546 TI - [Changing trends in HIV epidemics: recent diagnoses in the Spanish VACH cohort (2000-2002)]. AB - BACKGROUND: Studying the changing trends of HIV epidemics is a useful means of evaluating the results of current preventive plans as well as of defining future needs and objectives. METHODS: We performed a cross-sectional study of the newly diagnosed cases of HIV infection included in the Spanish VACH cohort. New HIV cases were defined as those diagnosed between January 2001 and December 2002. Their epidemiologic characteristics were compared with those of patients included in the same cohort who had been diagnosed between January 1998 and December 2000. RESULTS: We studied 603 new cases (27% women). In 146 (24.4%) HIV infection had been acquired by sharing material for intravenous drug use (IVDU), 171 (28,6%) were men who had had sex with other men (MSM) and 247 (41.3%) acknowledged some risk for heterosexual HIV transmission. The median age was 36 years (range: 18 80). Only 1.5% of the patients were younger than 20 years while 32.1% were older than 40 years. This percentage was significantly higher than that corresponding to 1998-2000 (27.5%; p < 0,05). HIV infection was diagnosed simultaneously with an AIDS-defining condition in 13.3% of patients and an AIDS-defining disease was diagnosed in the first month after HIV-diagnosis in another 40 patients (6.6%). CONCLUSIONS: We confirm the trends observed in previous studies: a growing proportion of newly diagnosed cases among women, a decreasing proportion of IVDU, a growth of MSM, and a trend toward diagnosis at a later age. PMID- 15826547 TI - Diagnosis of Enterocytozoon bieneusi by the polymerase chain reaction in archival fixed duodenal tissue. AB - This study involved ninety five formalin-fixed paraffin-embedded duodenal biopsy specimens retrieved from hospital files that were microscopically observed for the presence of microsporidia. Eleven samples that revealed compatible organisms were analyzed by the polymerase chain reaction (PCR) with four different protocols for the detection of Enterocytozoon bieneusi. Amplicons of the right size were obtained by at least one method for nine samples, remaining two negative ones. We report a PCR methodology that allows the use of archival specimens obtained for traditional pathology. PMID- 15826548 TI - [Recommendations from GESIDA/SEFH/PNS to improve adherence to antiviral treatment (2004)]. AB - Since the early days of antiretroviral therapy, adherence has emerged as the milestone of success; in fact, it is the most potent predictor of effectiveness. The main factors related to adherence include the complexity of the therapeutic regimen, adverse effects, psychological problems, alcoholism and active addiction to drugs, lack of social and family support and the patient's beliefs and attitudes about the treatment. Adherence monitoring should be part of the HIV patient's regular care, and should be done with feasible, easily applied methods adapted to the different clinical settings. The minimally acceptable measures should include use of a validated questionnaire, together with data from the Pharmacy Department's drug dispensation registry. All patients that begin HAART or undergo a change of treatment should participate in a treatment education program imparted by health professionals with knowledge and experience in the management of patients with HIV infection. The health team (doctors, pharmacists and nursing professionals) should offer maximum availability to solve the doubts and problems that may occur during treatment. When sub-optimal adherence is detected, intervention strategies based on psychological therapy, educational efforts and personal advice should be attempted, in order to adapt the treatment scheme to the patient's habits and provide solutions to the problem of non compliance. In certain situations, co-morbid conditions will also require attention. Treatment adherence, being a multidimensional problem, needs a multidisciplinary team approach. The choice of therapy, only one aspect of the multidimensional problem of adherence, must be a careful and individualized decision; however, simpler regimens with regard to the number of pills and daily dose are desirable. PMID- 15826549 TI - [Diseases produced by Borrelia]. AB - Lyme borreliosis, caused by Borrelia burgdorferi sensu lato, is a multi-organ infection with dermatological, rheumatological, neurological, and cardiac manifestations. The main characteristic is a skin lesion, named erythema migrans. Relapsing fever, caused by numerous species of Borrelia, is characterized by a periodic cycle of acute and afebrile episodes. The serological diagnosis of these infections has limited value in sensitivity, specificity and predictive values. Lyme borreliosis is usually diagnosed by recognition of a characteristic clinical picture with serological confirmation, and the diagnosis of relapsing fever relies on direct observation of spirochetes in peripherical blood. The elected treatment is almost always tetracycline for the young or for adults but not for pregnant women, although betalactamic (such as penicillin or 3rd generation cephalosporin for the central nervous system) or macrolides are indicated in several situations. The prognosis, with adequate treatment, is good. In the majority of Spanish regions, due to the low incidence of these diseases, the prophylactic antimicrobial treatment after a tick bite is not indicated. PMID- 15826550 TI - [Oral pain and loss of teeth in a 36-year-old woman]. PMID- 15826551 TI - [Slowly progressing skin ulcers following a stay in Costa Rica]. PMID- 15826552 TI - [Necrotizing fasciitis and bacteriemia]. PMID- 15826553 TI - [Meningeal cryptococcosis as a sign of immune reconstitution syndrome]. PMID- 15826554 TI - [Toxic epidermal necrolysis associated with abacavir]. PMID- 15826555 TI - [Brucellosis with multiple pulmonary nodules and optic neuritis]. PMID- 15826556 TI - [Severe community-acquired pneumonia due to Proteus mirabilis]. PMID- 15826557 TI - [Invasive meningococcal disease]. PMID- 15826558 TI - [Growth in healthy infants aged 0-2 years and comparison with reference charts]. AB - BACKGROUND: Growth reference charts should be periodically adapted to the population in which they will be applied, according to ethnic variation, secular growth change, and current feeding patterns. OBJECTIVE: To perform an anthropometric analysis of healthy contemporary infants aged 0-24 months and to compare the results with the most commonly used reference values. PATIENTS AND METHODS: We performed an observational, longitudinal and retrospective study of weight and length measured in the routine health checks of all infants born between 1998 and 2001 in the same Health Area and who received the infant formula beikost at a similar age. Exclusion criteria were low-birth-weight (< 2,500 g), prematurity, and chronic disease. Infants who did not complete the follow-up until the age of 2 years were also excluded. We calculated the means for each age, sex and variable. The Z-score represented by these means in the longitudinal charts most commonly used in Spain (Fundacion Orbegozo) and in the Euro-Growth study were calculated and were compared with Student's one-sample t-test. RESULTS: Of the 256 infants born in the study period, 30 were excluded (11.71%) and 226 infants were finally enrolled (52.2% boys). In relation to the Spanish reference values, in boys the weight Z-score at 12 and 24 months was +0.28 (95% CI: +0.10/+0.45) and +0.33 (95% CI: +0.15/+0.52) respectively, and the length Z score was +0.59 (95% CI: +0.43/+0.76) and +0.52 (95% CI: +0.33/+0.72). In girls the weight z-score was similar to that for boys (+0.24 [95% CI: +0.08/+0.40] and +0.37 [95% CI: +0.17/+0.57]) but growth in length was greater (+0.86 [95%CI: +0.66/+1.06] and +0.73 [95% CI: +0.52/+0.94]). This difference was statistically significant in all cases. At the age of 2 years, 17.75 % of girls and 8.47 % of boys were above the 97th percentile in length. No differences in length at 2 years were found in either boys or girls in relation to the Euro-Growth study. CONCLUSIONS: Anthropometric measurements in the first 2 years of life showed a significant increase in relation to the charts designed 20 years ago, especially in length and in girls. PMID- 15826559 TI - [Postnatal growth up to 2 years of corrected age in a cohort of very low birth weight infants]. AB - OBJECTIVE: To study postnatal somatic growth up to 2 years of corrected postnatal age (CPA) in a cohort of very low birth weight infants (VLBWI) in our unit, according to their intrauterine growth status at birth. PATIENTS AND METHODS: We performed an observational follow-up study of postnatal growth in a cohort of VLBWI (birth weight < 1,500 g) attended from January 1, 1994 to December 31, 1999. Weight, length, and head circumference were assessed at birth and again at 2 years CPA. Infants were classified as small-for-gestational age (SGA) if weight was below 2 standard deviations (SD) of the reference population mean. RESULTS: The cohort consisted of 260 VLBWI (18.5% SGA). Mean gestational age (SD) was 29 (2.7) weeks, weight was 1,150 (223) g, length was 37.4 (2.7) cm, and head circumference was 27 (1.8) cm. At birth Z-scores were -0.82, -0.92 and -0.8 for weight, length and head circumference. The corresponding mean Z scores at 2 years' CPA were -1.44, -1.49 and -0.31. At 2 years' CPA, more SGA infants still had a weight below the 3rd percentile than those with normal birthweight (26% vs. 16%). SGA infants had a higher daily weight increase during their hospital stay (27.5 vs. 21.2 g/day, p > 0.5). Gender did not significantly influence the growth pattern. CONCLUSIONS: Weight showed the lowest postnatal increase, particularly in infants < 1,000 g. Head circumference showed the greatest catch-up growth pattern, almost reaching the mean for the general population by 2 years' CPA. Growth retardation in SGA continued up to 2 years' CPA. PMID- 15826560 TI - [Morbidity at 2 years in infants with a birth weight of < 1,500 g]. AB - OBJECTIVES: To determine morbidity at 2 years' corrected age in a cohort of neonates with a birth weight of less than 1,500 g born in 1999-2000 in a tertiary hospital in the Autonomous Community of Madrid. PATIENTS AND METHODS: An observational longitudinal follow-up study was performed in a cohort of 213 infants with a birth weight of < 1,500 g. Of these, 188 (87%) completed the 2 year follow-up. Various types of neurosensorial disability were studied, paying special attention to the main impairments: vision, hearing and motor impairment, and low development quotient. RESULTS: Among the patients initially included in the study, 87% completed the follow-up; 17.1% had one or more major sequela. We found one case (0.4%) of bilateral neurosensorial deafness and one case of bilateral blindness. At the age of 2 years, 5.8 % (11/188) had cerebral palsy, 14.9% had a development quotient below 85 and 18% had not reached the 3rd percentile for weight. Factors of poor neurological prognosis were subnormal head size at the age of 2 years and white matter disease (including persistent intraparenchymal periventricular echodensity and ventriculomegaly or irregular shape) as ultrasound findings. CONCLUSIONS: Less than a fifth of the very low birth weight infants presented severe sequelae at 2 years of follow-up. Factors of poor neurological prognosis were subnormal head size at 2 years and the presence of white matter disease on ultrasonography. The findings on growth and development were worrying, since 18 % of the patients had not reached the 3rd percentile for weight at 2 years' corrected age. PMID- 15826561 TI - [Utility of procalcitonin, C-reactive protein and leukocyte count in the detection of meningococcal disease during an epidemic cluster]. AB - OBJECTIVE: To determine the clinical utility of biological markers of the acute phase of bacterial infection (procalcitonin, C-reactive protein and leukocyte count) in invasive meningococcal disease during an epidemic cluster. PATIENTS AND METHODS: Thirty-six patients with feverish syndrome who visited the emergency unit of our hospital within a 6-month period were studied. In all patients, serum procalcitonin and C reactive protein levels and leukocyte count were determined, and blood culture was performed. RESULTS: Invasive meningococcal disease, confirmed by blood culture, was found in seven of the 36 patients studied. The most frequent clinical presentation was a feverish syndrome of less than 24 hours of onset, progressing to sepsis in subsequent hours. Comparison of procalcitonin and C reactive protein concentrations in patients with and without meningococcal disease revealed that procalcitonin and C reactive protein levels greater than 10 ng/ml and 49.95 ng/ml respectively had high sensitivity, specificity and predictive values. Procalcitonin levels < 0.5 ng/ml were useful to easily rule out invasive meningococcal disease. In the case of leukocyte count, no value with clinical significance could be established, although counts were higher in patients with invasive meningococcal disease. CONCLUSIONS: The finding of prolactin levels of < 0.5 ng/ml and/or C-reactive protein levels of < 49.95 ng/ml in children or teenagers with fever of less than 24 hours of onset indicates a low probability of invasive meningococcal disease in epidemic situations. PMID- 15826562 TI - [Increasing diagnosis of eosinophilic esophagitis in Spain]. AB - INTRODUCTION: Eosinophilic esophagitis is a chronic inflammation of the esophagus characterized by marked eosinophilic infiltration. It is frequently associated with other allergic diseases. In the last few years, the incidence of eosinophilic infiltration has shown a striking increase in Spain, partly due to better knowledge of the disease and to the correct diagnosis of patients previously thought to be suffering from gastroesophageal reflux. MATERIAL AND METHODS: To report our experience in the diagnosis of eosinophilic esophagitis in the last few years and compare our data with previously published findings and current knowledge of the subject. RESULTS: From January 1997 to November 2003, 11 patients (nine boys and two girls) were diagnosed with eosinophilic esophagitis. The mean age at diagnosis was 9 years and 7 months. The most frequent symptoms were dysphagia with solids and food impaction. Seven patients had a familial history of allergy. Allergic tests were positive in eight patients; five had food allergy, five were positive for aeroallergens and two patients had mixed allergy. Endoscopy showed esophageal trachealization in two patients and papular whitish exudate in a further two; both these findings are characteristic of eosinophilic esophagitis. Endoscopic appearance was normal in seven patients. Esophageal biopsies showed > 20 eosinophils/hpf. Five patients had eosinophilic infiltration in other parts of the digestive tract. All the patients with food allergy were put on exclusion diets. Three patients received systemic steroids and cromolyn sodium and three received montelukast, with good response in all patients. CONCLUSIONS: The incidence of eosinophilic esophagitis is increasing. This disease should be considered in patients with longstanding symptoms presumed to be caused by gastroesophageal reflux or motility disorders with poor response to standard therapy. Eosinophilic esophagitis is frequently associated with allergy. A normal appearance of esophageal mucosa on endoscopy should not prevent the clinician from taking multiple biopsies. Patients with eosinophilic esophagitis show good response to anti-allergic treatment. PMID- 15826563 TI - [Case-control study of risk factors associated with constipation. The FREI Study]. AB - BACKGROUND: Children represent one of the patient groups most affected by constipation. Our objective was to identify and describe the risk factors associated with childhood constipation. PATIENTS AND METHODS: The study had a case-control, retrospective, open and multicenter design. Clinical data on possible risk factors were collected through an ad-hoc questionnaire. Two groups were studied: children with and without constipation. RESULTS: Nine hundred twenty-one children were recruited; of these, 898 (97.6%) were included in the statistical analysis. There were 408 (45.4%) children in the constipated group and 490 (54.5%) in the non-constipated group. Most of the children with constipation (53.6%) had a maternal history of constipation compared with 21.4% of children without constipation (p < 0.05). More than half (53.2%) of the constipated children reported a lack of regularity in their toilet habits while 64.9 % of the children without constipation went to the toilet regularly. Toilet training started slightly earlier (at 3 years) in children without constipation (93.2%) than in those with the disorder (83.8%) (p < 0.05). At school, 57.4% of the children with constipation never used the toilet compared with 26.8% of those without constipation (p < 0.05). A total of 73.4% of children with constipation drank less than four glasses of water per day compared with 47.1% of those without constipation (p < 0.05). Consumption of vegetables and legumes in the diet was significantly lower in children with constipation than in those without (p < 0.05). The risk factors linked to childhood constipation found in this study were a familial history of constipation, irregular toilet habits, low dietary fiber contents and no fruit intake. The main preventive factors against constipation were water and vegetable consumption and training on the use of the toilet at school. CONCLUSIONS: Daily toilet training and dietary changes are needed to prevent constipation among children and to achieve regular defecation. This preventive intervention should be reinforced at school. PMID- 15826564 TI - [Brainstem dysgenesis: Mobius, Cogan and Pierre Robin syndromes revisited]. AB - We propose the term brainstem dysgenesis to designate patients with congenital dysfunction of the cranial nerves and muscle tone due to prenatal lesions or anomalies of the brainstem. In some patients, the dysgenesis is genetically determined and can be isolated or form part of a more extensive polymalformation syndrome (mutations of organizing or regulatory genes). In most patients with brainstem dysgenesis, however, the disorder is caused by prenatal destructive or disruptive lesions of vascular origin. Depending on the vessels involved and the magnitude of the lesion, clinical manifestations can range from intrauterine death to mild involvement of several cranial nerves. Clinical findings in some of these patients may coincide with those described in Mobius, Pierre Robin or Cogan syndromes and, if that is the case, the eponym will indicate the location of the brainstem lesion. Clinical manifestations in most patients with brainstem dysgenesis, however, do not fit into any of the aforementioned syndromes. In these circumstances the term brainstem dysgenesis should be used followed by a detailed description of each patient's clinical findings and/or the brainstem segment presumably involved. The prognosis of patients with brainstem dysgenesis due to prenatal destructive lesions depends on the magnitude of the vascular territory involved and, in most cases, is better than the initial clinical manifestations would indicate. PMID- 15826565 TI - [Flexible bronchoscopy techniques: bronchoalveolar lavage, bronchial biopsy and transbronchial biopsy]. AB - This article completes previous recommendations of the Techniques Group of the Spanish Society of Pediatric Pulmonologists on the practice of flexible bronchoscopy in children. We review the most frequently performed diagnostic and therapeutic procedures applied through the flexible bronchoscope: bronchoalveolar lavage, bronchial biopsy and transbronchial biopsy. Recommendations are also provided on the practice of nonbronchoscopic bronchoalveolar lavage. We review the indications and contraindications of these techniques, the equipment required, and the preparation and monitoring of the patient before, during and after the procedure. The complications of these techniques are also discussed. These recommendations may be adopted, modified or rejected according to clinical needs and constraints. PMID- 15826566 TI - [Tuberous sclerosis associated with rhabdomyoma in the left ventricular outlet tract]. AB - We report a 2-month old newborn with a family history of adenoma sebaceum, achromic spots and renal lithiasis. Echocardiography-magnetic resonance imaging showed a solitary pediculated rhabdomyoma in the left ventricular outlet tract causing a severe systolic aortic valve obstruction. Surgical excision was carried out through the aortic valve with no valvular lesions. The patient presented multiple seizures 24 hours after surgery with good response to medical therapy. Ten days later, computed tomography showed a spot lesion in the subependimary area in the posterior position of the parietal horn, compatible with a hamartoma associated with tuberous sclerosis. PMID- 15826567 TI - [Odontogenic fibromyxoma]. AB - A 3-year-old boy presented a submandibular mass that progressively increased in size. Pathological study confirmed an odontogenic fibromyxoma. This entity is rare in children. PMID- 15826568 TI - [Mobius syndrome and an apparently life-threatening event]. AB - Mobius syndrome is characterized by congenital facial weakness with impairment of ocular abduction. A subgroup of these patients have associated apneas because of involvement of brainstem respiratory centers located slightly lateral to the abducens nuclei. We report a 7-month old infant admitted to the pediatric intensive care unit because of an episode of cyanosis, hypotonia and unresponsiveness. The patient then became respirator dependent afterwards. On examination, facial diplegia, impairment of ocular abduction and hypotonia were evident. Magnetic resonance imaging (MRI) revealed abnormal signal intensity in brainstem tegmentum. At the age of 11 months he was discharged but required a home ventilator. He died 5 months later due to an infection. Mobius syndrome is associated with central respiratory dysfunction. The finding of abnormal signal intensity in brainstem tegmentum on MRI is a possible predictor of apnea in these patients. PMID- 15826569 TI - [Adverse reaction of pseudoephedrine]. AB - We present a case of a 7 years old girl who developed an episode of myoclonic movements and tremors after being medicated with a not well quantified amount of a pseudoephedrine/antihistamine combination. We want to highlight the potential toxicity of pseudoephedrine, usually administered as part of cold-syrup preparations which are used for symptomatic treatment of upper respiratory tract cough and congestion associated with the common cold and allergic rhinitis. Although these products are generally considered to be safe either by physicians and parents, we can't underestimate the potential adverse events and toxic effects that can occur when administering these medications. PMID- 15826570 TI - [Reasons for the 2003-2004 meningitis C vaccination campaign in Castilla-La Mancha]. PMID- 15826571 TI - [Solid cardiac mass due to hydatidosis]. PMID- 15826573 TI - [A rare lesion in the pediatric population: Mucinous cystoadenoma of ovary]. PMID- 15826574 TI - [Asymptomatic congenital bronchial atresia in a 10-year-old boy]. PMID- 15826575 TI - [Use of helmets with bicycles and motorbikes]. PMID- 15826576 TI - [Thyroid hemiagenesia diagnosed in a 5-month-old infant]. PMID- 15826577 TI - [Lichen striatus]. PMID- 15826578 TI - [Hypohidrotic ectodermal dysplasia]. PMID- 15826579 TI - [Association between microsatellite instability and clinico-pathological characteristics in sporadic colon cancer]. AB - BACKGROUND AND OBJECTIVE: Currently, colon cancer is a leading cause of cancer death world-wide. It progresses according to three molecular pathways, named suppressor, mutador and methylator. Microsatellite instability is a hallmark of the lack of reparation, of DNA mismatches and it characterizes a subset of colon tumors (unstable tumors, MSI). MSI-H patients (high degree of microsatellite instability) seem to share clinico-pathological differences with MSS (microsatellite stable) and MSI-L (low degree of microsatellite instability) patients. In this study, associations between high degree of microsatellite instability and pathological (location, mucinous content, differentiation grade, stages T3N0, stages II and III) and clinical features (response to chemotherapy, disease-free survival and overall survival) were evaluated. PATIENTS AND METHOD: 117 patients with sporadic colon cancer were classified into two populations (MSS/MSI-L and MSI-H) by using PCR and electrophoresis of seven microsatellites, according to the National Cancer Institute recommendations. RESULTS: MSI-H tumors tended to be located in the right colon (p = 0.022) and were of mucinous histologic type (p = 0.04). No differences in disease-free survival and overall survival between group of stage II and III patients with MSS/ MSI-L and corresponding ones with MSI-H colon cancer were found (p = 0.54, p = 0.37, respectively). Conversely, MSI-H patients with stage II colon cancer had a favourable prognosis (p = 0.027). Nevertheless, response to 5-fluorouracil (5-FU) and leucovorin was similar in MSS/ MSI-L and MSI-H groups (p = 0.38). CONCLUSIONS: MSI-H patients are characterized by certain pathological features; those MSI-H patients with a stage II seem to have a better prognosis than MSS/ MSI-L patients. PMID- 15826580 TI - [Influence of comorbidity on admission management and pharmacological treatment prescribed at discharge in acute myocardial infarction]. AB - BACKGROUND AND OBJECTIVE: The management of cardiac ischemic patients differs depending on their comorbidity. The Charlson Index (ChI) and its adaptations are well established and widely used tools to quantify a patient comorbidity. The aim of this study is to evaluate the influence of comorbidity quantified by the ChI in the treatment administered at admission and in the pharmacological treatment prescribed at discharge in the setting of an acute myocardial infarction with and without ST segment elevation. PATIENTS AND METHOD: We studied a total of 955 patients consecutively admitted in our hospital with the diagnosis of acute myocardial infarction. Comorbidity was obtained at the first day of admission applying the ChI. According to this value patients were classified from minor to major in 2 subgroups (ChI or= 2) and differences in the admission and discharge treatments between both groups were analyzed. RESULTS: Patients admitted with acute myocardial infarction without ST segment elevation and ChI > 2 received less frequently betablockers at discharge, but there were no significant differences in the use of ACE inhibitors, calcium channel blockers or statins. In addition they were submitted less frequently to revascularization procedures or treadmills, and no differences were found in the use of echocardiograms. Patients with ST segment elevation and ChI > 2 were less frequently treated with betablockers or statins at discharge, and were submitted to less treadmills or echocardiograms; furthermore, in these patients, there were no significant differences in the use of ACE inhibitors, calcium channel blockers, thrombolytics or revascularization procedures. CONCLUSIONS: Comorbidity quantified on admission by the ChI is an independent factor that modifies in hospital and ambulatory management of patients with acute myocardial infarction. There is a lower use of invasive techniques as well as a lower prescription of betablockers at discharge in patients with greater comorbidity. PMID- 15826581 TI - [Chronic fatigue syndrome and multiple chemical hypersensitivity after insecticide exposure]. AB - BACKGROUND AND OBJECTIVE: Chronic Fatigue Syndrome (CFS) and Multiple Chemical Sensitivity (MCS) are well-defined illnesses that may appear after some toxic exposures. PATIENTS AND METHOD: We report a consecutive series of 26 patients who developed CFS after exposure to insecticide products. It was associated with MCS in a third of cases. RESULTS: Toxic exposure was of labour origin after returning to usual work place after a process of fumigation. In 42% of cases there was no fulfilment of fumigation safety rules. The majority of patients were mean-aged women who developed an acute upper airway inflammatory syndrome, without muscarinic or nicotinic manifestations, followed by digestive syndrome, neurocognitive, fibromyalgic and chronic fatigue manifestations. The course of disease was shorter than 1 year in 5 cases (19%), longer than 1 year in 15(58%), and disabling in 6 cases (23%). CONCLUSIONS: Due to the possible prevention of this toxic exposure, it is very important to carefully follow measures of environment isolation and ventilation after insecticide use in order to avoid the development of these diseases. PMID- 15826582 TI - [Colorectal cancer and microsatellite instability]. PMID- 15826583 TI - [From Hippocrates to genomics in clinical medicine]. PMID- 15826584 TI - [Chronic open angle glaucoma]. PMID- 15826585 TI - [Role of calcium intake in obesity]. AB - Recent evidences from epidemiological and intervention trials suggest that a higher calcium intake is associated with a lower body weight and a reduction in total and central body fat. The potential role of calcium intake in the regulation of body adiposity has been explained by the intracellular calcium, which is able to promote adipocyte fat accumulation by exerting a coordinate regulation stimulating lipogenesis and suppressing lipolysis. Moreover, the intracellular calcium has been implicated on the etiopathogenesis of hypertension, insulin resistance and dyslipidemia, appearing as a potential common substrate in the metabolic syndrome. Dietary modulation of intracellular calcium through calciotropic hormones opens an exciting possibility to prevent and tackle obesity and its associated complications by increasing the calcium intake. However, available evidences are mostly indirect and further studies specifically designed for this purpose are needed. PMID- 15826586 TI - [A way to make clinical pragmatism operative: sistematization of the actuation of competent physicians]. PMID- 15826588 TI - [Epidemiology of asymptomatic pulmonary embolism in cancer patients]. PMID- 15826589 TI - [Liver fibrosis in HIV/HCV co-infected patients]. PMID- 15826590 TI - [Palivizumab and bronchiolitis: need to apply the efficiency in order to join clinical practice to scientific studies]. PMID- 15826592 TI - [Massive brain hemorrhage in a case of Creutzfeldt-Jakob disease]. PMID- 15826600 TI - 13-cis retinoic acid and isomerisation in paediatric oncology--is changing shape the key to success? AB - Retinoic acid isomers have been used with some success as chemotherapeutic agents, most recently with 13-cis retinoic acid showing impressive clinical efficacy in the paediatric malignancy neuroblastoma. The aim of this commentary is to review the evidence that 13-cis retinoic acid is a pro-drug, and consider the implications of retinoid metabolism and isomerisation for the further development of retinoic acid for cancer therapy. The low binding affinity of 13 cis retinoic acid for retinoic acid receptors, low activity in gene expression assays and the accumulation of the all-trans isomer in cells treated with 13-cis retinoic acid, coupled with the more-favourable pharmacokinetic profile of 13-cis retinoic acid compared to other isomers, suggest that intracellular isomerisation to all-trans retinoic acid is the key process underlying the biological activity of 13-cis retinoic acid. Intracellular metabolism of all-trans retinoic acid by a positive auto-regulatory loop may result in clinical resistance to retinoic acid. Agents that block or reduce the metabolism of all-trans retinoic acid are therefore attractive targets for drug development. Devising strategies to deliver 13-cis retinoic acid to tumour cells and facilitate the intracellular isomerisation of 13-cis retinoic acid, while limiting metabolism of all-trans retinoic acid, may have a major impact on the efficacy of 13-cis retinoic acid in paediatric oncology. PMID- 15826601 TI - Bis(pivaloyloxymethyl) thymidine 5'-phosphate is a cell membrane-permeable precursor of thymidine 5'-phosphate in thymidine kinase deficient CCRF CEM cells. AB - Bis(pivaloyloxymethyl) thymidine 5-phosphate (POM(2)-dTMP) has been investigated as a membrane-permeable prodrugs of dTMP. The growth inhibitory activity of POM(2)-TMP has been compared with thymidine (TdR) in wild type CCRF CEM cells (CEM) and a strain that lacks TdR kinase (CEM tk-). After 72 h incubation at 37 degrees C, TdR showed significant antiproliferative activity (IC(50)=27 microM) against CEM cells but was weakly effective (IC(50)=730 microM) against the mutant cell line. By comparison, bis(pivaloyloxymethyl) thymidine 5'-monophosphate (POM(2)-dTMP) was equally inhibitory (IC(50)=5 microM) to both cell lines. The growth inhibitory effects were reversed by deoxycytidine. Cellular [methyl (3)H]dTTP pools increased linearly over 2h during incubation of CEM or CEM tk- with 5 microM POM(2)-[methyl-(3)H]dTMP. The incorporation of [methyl-(3)H]TdR into HClO(4)-insoluble cell residue by CEM tk- was <0.1% that of CEM and did not increase over 1h. In contrast, CEM tk- incorporated radioactivity from POM(2) dTMP into acid insoluble residue at a rate 59% that of CEM. These results demonstrate that POM(2)-dTMP can penetrate into cells and serve as a source of dTMP. PMID- 15826602 TI - Involvement of multiple signaling pathways in the post-bariatric induction of IL 6 and IL-8 mRNA and release in human visceral adipose tissue. AB - The present studies were designed to determine the site of and the mechanism for the rapid increase in IL-6 and IL-8 mRNA observed in human visceral adipose tissue after removal during laparoscopic bariatric surgery. Upregulation of IL-6 and IL-8 mRNA as well as their release were seen within 3h whether one intact piece of tissue or minced pieces of adipose tissue were incubated in vitro. Most of the IL-6 and IL-8 mRNA content of visceral adipose tissue after 3h of incubation was in the non-fat cells. Actinomcyin D markedly reduced the upregulation of IL-6 and IL-8 mRNA. Incubation of adipose tissue explants with a soluble TNFalpha receptor (etanercept) plus a blocking antibody against IL-lbeta reduced by 55% the increase in IL-6 mRNA and by 42% that of IL-8 mRNA seen between 1 and 5h of incubation. The upregulation of IL-8 and IL-6 mRNA accumulation as well as their release over a 2 or 4h incubation was reduced by around 50% in the presence of an inhibitor of the p38 MAPK or an inhibitor of the NFkappaB pathway and by 85% in the presence of both inhibitors. The data suggest that the relative trauma and/or hypoxia that occurs when adipose tissue is removed results in the release of TNFalpha and IL-1beta. These cytokines, and probably other factors as well, enhance IL-6 and IL-8 mRNA accumulation in human adipose tissue explants through mechanisms involving the p38 MAPK and NFkappaB pathways. PMID- 15826603 TI - Severity of pancreatitis-associated gut barrier dysfunction is reduced following treatment with the PAF inhibitor lexipafant. AB - The aim of the present study was to investigate the potential effect of treatment with a platelet-activating factor (PAF) antagonist, lexipafant (BB-882), on gut endothelial and epithelial barrier dysfunction and leukocyte recruitment in rats with acute pancreatitis. Severe acute pancreatitis was induced by the intraductal administration of 5% sodium taurodeoxycholate and pancreatitis-associated gut barrier dysfunction was characterized by increased exudation of radiolabelled albumin into the interstitium and alterations in bidirectional (over both the endothelial and epithelial barrier components) permeability of the intestine at the early stage of bile salt-induced acute pancreatitis. Levels of interleukin 1beta and 6, ileal and colonic myeloperoxidase (MPO) content, clearance of radiolabelled albumin from blood to the gut lumen or gut lumen to blood, and leakage of radiolabelled albumin to the ileum or colon were measured 3 and 12h after induction of acute pancreatitis. Treatment with lexipafant 30 min and 6h after pancreatitis reduced severity of pancreatitis-associated intestinal dysfunction, associated with a diminish in systemic concentrations of IL-1 and local leukocyte recruitment. The findings imply that PAF plays a critical role in the development of pancreatitis-associated gut barrier dysfunction and that PAF antagonist in some forms may represent potential candidates for future therapeutic intervention. PMID- 15826604 TI - G2 arrest and apoptosis by 2-amino-N-quinoline-8-yl-benzenesulfonamide (QBS), a novel cytotoxic compound. AB - We screened a library of 11,000 small molecular weight chemicals, looking for compounds that affect cell viability. We have identified 2-amino-N-quinoline-8-yl benzenesulfonamide (QBS) as a potent cytotoxic compound that induces cell cycle arrest and apoptosis. Treatment of Jurkat T cells with QBS increased the levels of cyclin B1 as well as phosphorylated-cdc2, which was accompanied by reduced activity of cdc2 kinase, suggesting that QBS may induce cell cycle arrest at G2 phase. Structural analogues of QBS also exhibited similar effects on cell cycle progression and cell viability. Long-term treatment with QBS resulted in DNA fragmentation, cytochrome C release, and PARP cleavage, and an increase in the number of subdiploidy cells, indicative of cellular apoptosis. Moreover, QBS induced apoptosis was blocked by z-VAD-fmk, a pan-caspase inhibitor. These results suggest that QBS is a novel and potent compound that induces G2 arrest and subsequent apoptosis, implicating it as a putative candidate for chemotherapy. PMID- 15826605 TI - The effect of quercetin on pro-apoptotic activity of cisplatin in HeLa cells. AB - It is well known that some tumour cells are very resistant to chemotherapy induced cell death which indicate poor prognosis for patients. Thus the aim of the present study was to investigate the effect of quercetin on pro-apoptotic activity of cisplatin in human cervix carcinoma cells (HeLa). Three variants of experiments were performed. In the first one cells were incubated with studied drugs separately for 8 and 24h. In the second, drugs were added to the culture medium simultaneously. In third cisplatin or quercetin addition was followed by subsequent quercetin or cisplatin treatment, respectively. We observed different apoptotic effects, dependent on the drug succession. Preincubation of cells with quercetin followed by cisplatin treatment appeared to be the most effective and was correlated with strong activation of caspase-3 and inhibition of both heat shock proteins (Hsp72) and multi-drug resistance proteins (MRP) levels. Our results indicate that quercetin pretreatment sensitizes HeLa cells to cisplatin induced apoptosis in HeLa cells. PMID- 15826606 TI - Very stable superoxide radical adducts of 5-ethoxycarbonyl- 3,5-dimethyl pyrroline N-oxide (3,5-EDPO) and its derivatives. AB - Oxygen radicals are involved in the onset of many diseases. Adequate spin traps are required for identification and localisation of free radical formation in biological systems. Superoxide spin adducts with half-lives up to 20 min at physiological pH have recently been reported to be formed from derivatives of the spin trap 5-ethoxycarbonyl-5-methyl-1-pyrroline N-oxide (EMPO). This is a major improvement over DMPO (t(1/2) ca. 45 s), and even DEPMPO (t(1/2) ca. 14 min). In this study, an additional methyl group was introduced into position 3 or 4 of the pyrroline ring which greatly increases the stability of the respective superoxide spin adducts. In addition, the ethoxy group of EMPO was exchanged by either a propoxy- or an iso-propoxy group in order to test the influence of increasing lipophilic properties of the investigated spin traps. The structure of all compounds was confirmed by (1)H and (13)C-NMR with full signal assignment. In comparison with EMPO (t(1/2) ca. 8 min) or DEPMPO (t(1/2) ca. 14 min), the superoxide adducts of all novel spin traps were considerably higher (t(1/2) ca. 12-55 min). In addition, various other spin adducts obtained from oxygen-centered as well as carbon-centered radicals (e.g. derived from methanol or linoleic acid hydroperoxide) were also detected. PMID- 15826607 TI - Clofibrate and perfluorodecanoate both upregulate the expression of the pregnane X receptor but oppositely affect its ligand-dependent induction on cytochrome P450 3A23. AB - The pregnane X receptor (PXR) interacts with a vast array of structurally dissimilar chemicals and confers induction of several major types of drug metabolizing enzymes such as cytochrome P450s (CYP). We previously reported that the expression of PXR was markedly increased in rats treated with clofibrate and perfluorodecanoic acid (PFDA). The present study was undertaken to test the hypothesis that induced expression of PXR increases PXR ligand-dependent induction on CYP3A23. Rat hepatocytes were treated with clofibrate or PFDA individually, or along with PXR ligand pregnenolone 16alpha-carbonitrile (PCN), and the levels of PXR and CYP3A23 were determined by Western blots. Both clofibrate and PFDA markedly increased the expression of PXR with PFDA being more potent, and the induction was abolished by actinomycin D, an inhibitor for mRNA synthesis. As expected, PCN alone markedly induced the expression of CYP3A23. Interestingly, co-treatment with clofibrate enhanced the induction, whereas co treatment with PFDA suppressed it. Clofibrate and PFDA represent multi-classes of chemicals called peroxisome proliferators including many therapeutic agents and industrial pollutants. The opposing effects of clofibrate and PFDA on the PCN induced expression of CYP3A23 suggest that peroxisome proliferators likely increase the expression of PXR but differentially alter its ligand-dependent induction. The interaction between PXR inducer and ligand provides a novel mechanism on how functionally and structurally distinct chemicals cooperatively regulate the expression of xenobiotic-metabolizing enzymes and transporters. PMID- 15826608 TI - The role of NMDA receptor upregulation in phencyclidine-induced cortical apoptosis in organotypic culture. AB - Phencyclidine (PCP) is an N-methyl-D-aspartate receptor (NMDAR) antagonist known to cause selective neurotoxicity in the cortex following subchronic administration. The purpose of this study was to test the hypothesis that upregulation of the NMDAR plays a role in PCP-induced apoptotic cell death. Corticostriatal slice cultures were used to determine the effects of NMDAR subunit antisense oligodeoxynucleotides (ODNs) on PCP-induced apoptosis and NMDAR upregulation. NR1, NR2A or NR2B antisense ODNs were incubated alone or with PCP for 48h. One day following washout, it was observed that PCP treatment caused an increase in NR1, NR2A and Bax polypeptides in the cortex, but had no effect on Bcl-xL. These increases were associated with an increase in cortical histone associated DNA fragments. Co-incubation of PCP with either NR1 or NR2A antisense significantly reduced PCP-induced apoptosis, while neither NR2B antisense ODN nor NR1 sense ODN used as a control had an effect. This effect was exactly correlated with the ability of the antisense ODNs to prevent PCP-induced upregulation of NR subunit proteins and the pro-apoptotic protein, Bax. That is, western analysis showed that antisense ODNs directed against either NR1 or NR2A prevented PCP induced increases in Bax in addition to preventing the upregulation of the respective receptor proteins. On the other hand, the NR2B antisense ODN had no effect on either NR2B protein or on Bax. These data suggest that NR1 and NR2A antisense ODNs offer neuroprotection from apoptosis, and that upregulation of the NR1 and NR2A subunits following PCP administration is at least partly responsible for the observed apoptotic DNA fragmentation. PMID- 15826609 TI - Reduced nicotine distribution from mother to fetal brain in rats vaccinated against nicotine: time course and influence of nicotine dosing regimen. AB - Nicotine is a teratogen in rats and possibly in humans. Vaccination against nicotine is being studied as a possible treatment for nicotine dependence. The safety of maternal vaccination against nicotine during or prior to pregnancy is not known. In this study, female rats were vaccinated and then administered acute or chronic nicotine during pregnancy at doses simulating nicotine exposure in smokers. Maternal vaccination reduced nicotine distribution to both maternal brain (44-47%) and fetal brain (17-39%) for up to 25 min after a single maternal nicotine dose administered on gestational day (GD) 20, but had a smaller effect on nicotine distribution to brain after continuous nicotine infusion. Nicotine distribution to maternal or fetal brain after repeated nicotine bolus doses was reduced immediately following an individual dose in vaccinated rats, but the chronic accumulation of nicotine in fetal brain was not altered. Nicotine distribution to whole fetus, in contrast to fetal brain, was generally not altered by vaccination. Nicotine-specific antibody concentration in fetal serum was 10% that of maternal serum, and in fetal brain was <1% of maternal serum. Although nicotine transfer to the whole fetus was not reduced by vaccination, protein binding data suggest that nicotine-specific antibody transferred from mother to fetus served to bind nicotine in fetal serum, reduce the unbound nicotine concentration, and thereby reduce nicotine distribution to fetal brain. These data comment on the safety of vaccination against nicotine during pregnancy, and suggest that vaccination may reduce the distribution of nicotine to fetal brain under some nicotine dosing conditions. PMID- 15826610 TI - Serotonin glucuronidation by Ah receptor- and oxidative stress-inducible human UDP-glucuronosyltransferase (UGT) 1A6 in Caco-2 cells. AB - Caco-2 cells are a widely used model in drug development to study intestinal drug transport and metabolism. Recently, serotonin (5-hydroxytryptamine, 5-HT) has been characterized as a highly selective substrate of human UDP glucuronosyltransferase UGT1A6 [Krishnaswamy S, Duan SX, von Moltke LL, Greenblatt DJ, Court MH. Validation of serotonin (5-hydroxytryptamine) as an in vitro substrate probe for human UDP-glucuronosyltransferase (UGT) 1A6. Drug Metab Disp 2003; 31:133-9], an isoform which conjugates planar phenols and is inducible by Ah receptor agonists and by oxidative/electrophile stress. To gain more insight into intestinal 5-HT disposition, uptake and metabolism of this neurotransmitter was studied in Caco-2 cell monolayers. It was found that 5-HT was taken up from the basolateral and to a lesser extent from the apical surface. It was mainly excreted basolaterally as 5-HT glucuronide. 5-HT UGT activity and UGT1A6 mRNA were induced by Ah receptor agonists and by oxidative stress generated by tert-butylhydroquinone and by isomeric thymoquinone, a potential antitumor agent and constituent of Nigella sativa seeds, commonly used as a condiment in the Middle East. While UGT1A6 induction was clearly detectable in NAD(P)H:quinone oxidoreductase 1 (NQO1)-deficient Caco-2 cells, it was not induced in NQO1-efficient HT-29 colon adenocarcinoma cells. The results suggest that--in addition to its detoxification function--intestinal UGT1A6 contributes to intestinal homeostasis of 5-HT from dietary sources and from release by enterochromaffin cells. PMID- 15826611 TI - Appearance questions can be misleading: a discourse-based account of the appearance-reality problem. AB - Preschoolers' success on the appearance-reality task is a milestone in theory-of mind development. On the standard task children see a deceptive object, such as a sponge that looks like a rock, and are asked, "What is this really?" and "What does this look like?" Children below 412 years of age fail saying that the object not only is a sponge but also looks like a sponge. We propose that young children's difficulty stems from ambiguity in the meaning of "looks like." This locution can refer to outward appearance ("Peter looks like Paul") but in fact often refers to likely reality ("That looks like Jim"). We propose that "looks like" is taken to refer to likely reality unless the reality is already part of the common ground of the conversation. Because this joint knowledge is unclear to young children on the appearance-reality task, they mistakenly think the appearance question is about likely reality. Study 1 analyzed everyday conversations from the CHILDES database and documented that 2 and 3-year-olds are familiar with these two different uses of the locution. To disambiguate the meaning of "looks like," Study 2 clarified that reality was shared knowledge as part of the appearance question, e.g., "What does the sponge look like?" Study 3 used a non-linguistic measure to emphasize the shared knowledge of the reality in the appearance question. Study 4 asked children on their own to articulate the contrast between appearance and reality. At 91%, 85%, and 81% correct responses, children were at near ceiling levels in each of our manipulations while they failed the standard versions of the tasks. Moreover, we show how this discourse based explanation accounts for findings in the literature. Thus children master the appearance-reality distinction by the age of 3 but the standard task masks this understanding because of the discourse structure involved in talking about appearances. PMID- 15826612 TI - Feature inference and the causal structure of categories. AB - The purpose of this article was to establish how theoretical category knowledge specifically, knowledge of the causal relations that link the features of categories-supports the ability to infer the presence of unobserved features. Our experiments were designed to test proposals that causal knowledge is represented psychologically as Bayesian networks. In five experiments we found that Bayes' nets generally predicted participants' feature inferences quite well. However, we also observed a pervasive violation of one of the defining principles of Bayes' nets-the causal Markov condition-because the presence of characteristic features invariably led participants to infer yet another characteristic feature. We argue that this effect arises from a domain-general bias to assume the presence of underlying mechanisms associated with the category. Specifically, people take an exemplar to be a "well functioning" category member when it has most or all of the category's characteristic features, and thus are likely to infer a characteristic value on an unobserved dimension. PMID- 15826614 TI - Development of pre-hospital trauma-care system--an overview. AB - Trauma is recognized as a serious health care problem world-wide and there is a general agreement that trauma is best addressed in a systematic manner. Accordingly, trauma systems must be designed to use efficiently the limited health care resources available. The majority of injuries are of only minor, or moderate, severity, and can be managed well at local community hospitals. On the other hand, a significant minority of injured patients will require extensive and expensive care in order to survive, or to minimize the morbidity of injury. For a trauma system to be optimally effective, given limited available resources and the need for urgent care of the seriously injured, it is critical to utilize a method of differentiating those injury victims, who need the specialized expertise and resources available in trauma centres, from those who can be cared for adequately locally. Rapid field assessment of injured patients is, therefore, an essential element of any effectively functioning trauma system. This review examines the evolution of pre-hospital trauma-care systems, during the triage of a trauma victim. PMID- 15826615 TI - Where to split plaster casts. AB - Plaster casts are often split to accommodate swelling following injury. This is not influenced by the axis of the split. The aim of this study was to compare the mechanical properties of plasters split along different axes. Full plasters were applied to mannequin forearms, and then split along dorsal, volar, radial or ulnar sides. Following this the plasters were loaded in a dorsal direction. We found that of all the axes tested, the dorsal split was the best for maintaining fracture reduction (p = 0.001). PMID- 15826616 TI - Complications encountered while using thin-wire-hybrid-external fixation modular frames for fracture fixation. A retrospective clinical analysis and possible support for "Damage Control Orthopaedic Surgery". AB - One hundred ninety eight adult patients who had sustained long bone fractures were treated by external fixation from admission to bone healing and consolidation. Of these, 135 had sustained high-energy injuries, 39 of them had suffered multi-system injuries. Superficial pin track infection was the most common complication, occurring predominantly in pins located in the femur, upper tibia and upper humerus. There were no cases of deep infection or osteomyelitis. One patient with a femoral shaft fracture developed a DVT although he was on preventive low molecular weight heparin, i.e. sc Clexane 40 mg daily. There were no cases of PE or ARDS. External fixation systems are a minimal invasive surgical modality, which allow three-dimensional fracture fixation after closed or minimal open reduction. They require a good command of surgical anatomy, but provide an optimal preservation of the fracture's soft tissue envelope, the critical biological factor for new bone formation and fracture healing. Recent publications have suggested that in the critically ill patient, minimally invasive fracture fixation surgery may prevent the perpetuation of a reactive, life threatening inflammatory reaction (the "second hit") which may induce the development of multiple organ dysfunction (MODS). PMID- 15826617 TI - Complications of temporary and definitive external fixation of pelvic ring injuries. AB - OBJECTIVES: To determine the incidence of complications of external fixation in pelvic ring injuries, comparing fixator use for temporary and definitive treatment. DESIGN: Retrospective case-note review. SETTING: A regional centre for pelvic trauma in the UK. PATIENTS: 100 consecutive patients. INTERVENTION: All patients were treated with pelvic external fixation for a pelvic ring injury. RESULTS: In 52 patients, external fixation was intended for use as the definitive treatment of the pelvic ring injury and was maintained for a mean duration of 60 days (17-113). In 48 patients, it was used temporarily for a mean duration of 8 days (1-20) before internal fixation of the pelvic ring. The complication rate for definitive and temporary fixators was 62 and 21%, respectively. Pin-site infection occurred in 50% of definitive fixators and 13% of temporary fixators but rarely led to more serious complications. In five patients, the definitive management was changed as a result of a complication of the external fixator. The commonest cause for revision of either fixator was aseptic pin loosening. Revision for loose pins in eight patients was associated with the use of two pins in each iliac crest rather than three. CONCLUSIONS: The temporary use of external fixation is safe and effective, but use for definitive treatment is associated with a high rate of infection and aseptic pin loosening. PMID- 15826618 TI - Operative management of acetabular fractures. A review of 73 fractures. AB - Between June 1994 and December 2000, 80 displaced acetabular fractures were treated by open reduction and internal fixation, where possible by a single surgical approach. We report our results in 72 patients (73 fractures) who completed a minimum follow-up of 2 years. The average age of patients was 39.5 years (range 15-76 years). Thirty-four fractures were classified as simple and the remaining 39 were complex fractures. Twenty-four patients were recorded to have posterior dislocation of the hip joint at the time of initial presentation and at surgery eight patients were noted to have varying degrees of damage to the head of the femur. The average time to surgery was 11.7 days (range 1-35 days); 80% of cases were operated within 2 weeks period. The average follow-up was 45.5 months (range 24-96 months). In 67 fractures (92%), only a single approach was used (Anterior Ilioinguinal 26 cases; Posterior Kocker-Lagenbach 41 cases). Five fractures needed an extensile triradiate approach and only one case required a combined anterior and posterior approach. Congruent reduction was achieved in 89% of cases. Main complications included deep infection in two patients (2.7%) and severe degenerative changes in three patients (4.2%). A total of four patients (5.5%) required a total hip replacement. There were no cases of deep vein thrombosis or pulmonary embolism. Heterotopic ossification grade III was seen in three (4.1%) cases and none were grade IV. At the latest follow-up the average Harris hip score was 85 (range 20-100). We conclude that in the majority of cases internal fixation is possible through a single approach provided patients are referred early to a specialist unit. This results in satisfactory outcome with reduced morbidity and complication rate. PMID- 15826619 TI - Impact energy absorption by specimens from the upper end of the human femur. AB - A cadaveric biomechanical study was performed to investigate the fracture energy absorbed by strips of bone from the proximal femur in relation to age and gender, under impact loading conditions. Four groups (young male, young female, old male, old female) of four cadaveric proximal femurs were used in each case. Four bone strips were taken from the neck and four from the subtrochanteric area and these were tested under dynamic-impact conditions using the Charpy impact test. The fracture energy was calculated as the energy needed to achieve fracture per unit area, and expressed in J/m2. Bone specimens from young males are significantly tougher under impact conditions to those of females (p = 0.001), whereas between the old male and female groups, fracture energy does not significantly differ (p = 0.165). There was also significant difference (p < 0.0005) between the young and the old groups in both genders. The fracture energy absorption of the subtrochanteric area compared to that of the femoral neck for the same group of age and gender is in general slightly higher for all groups. In conclusion, gender in the young age group played a significant role in bone resistance in breaking whereas in the older age group it played a less important role. PMID- 15826620 TI - Dislocated Thompson hemiarthroplasty--the management of the recurrent dislocator. AB - Dislocation of a Thompson hemiarthroplasty of the hip is a serious complication with a high mortality rate. Previous papers have focused on surgical techniques to try and prevent dislocation. There is little in the literature on how to manage a patient after a dislocation. Patients with a dislocated Thompson hemiarthroplasty over a 5-year period from 1997 to March 2002 were analysed. Attempts were made to identify factors which may contribute to redislocation. Our strategies for preventing redislocation were evaluated. Of the 612 patients who received a Thompson hemiarthroplasty 23 patients (4%) dislocated. The average number of dislocations per patient was 2.4. Thirteen patients (57%) redislocated their prosthesis. Ten patients (43%) dislocated at least twice. Seven patients (30%) had either a total hip replacement, Girdlestone's procedure or the hip was left dislocated. Out of 15 patients fitted with an abduction brace 8 (60%) redislocated. Out of 8 patients treated with traction 6 (75%) redislocated. The 6 month mortality for patients suffering a dislocation was 7/23 (30%). If the prosthesis dislocates twice, the hip should be deemed unstable and consideration should be given to a revision procedure. Abduction braces and traction are ineffective in this condition and should be abandoned. PMID- 15826621 TI - Fracture of the femoral neck, the risk of serious underlying pathology. AB - Between 1997 and 2003, a total of 2223 patients presented to our hospital with the neck of femur fracture. This retrospective study examines the histological results of 90 patients (4%) suspected of having a fracture associated with serious underlying pathology. The mean age at the time of fracture was 80 years (44-97). The patients were divided into four groups. Group I: 34 patients with fracture without history of fall or trauma. Group II: 21 patients with suspicious lesions on the standard pelvic radiograph taken on admission. Group III: 27 patients with past history of malignancy without known bone metastases. Group IV: 8 patients with past history of malignancy and known bone metastases. None of the patients in groups I and II had significant abnormalities other than osteoporosis. Four patients (15%) in group III and six patients (75%) in group IV had metastases on histological examination. It is concluded that only groups III and IV are likely to have fractures due to metastatic disease. PMID- 15826622 TI - Varus impacted intracapsular hip fractures. AB - We describe a distinct variant of an impacted intracapsular hip fracture that is difficult to classify within any of the current fracture classification systems. Instead of impaction occurring laterally, as generally occurs with a Garden grade I fracture, there is medial impaction. We have termed this a varus impacted fracture. Nineteen such fractures were identified. The majority of these patients presented with a history of progressive hip pain over several days without a definite history of trauma. For the 16 fractures treated by internal fixation without any attempt at fracture reduction, fracture healing occurred in 11 cases. PMID- 15826623 TI - Role of valgus osteotomy and fixation by double-angle blade plate in neglected displaced intracapsular fracture of neck of femur in younger patients. AB - Head preservation is the mainstay of management in younger patients with intracapsular fracture neck of femur. In this prospective study, we have tried to evaluate the role of valgus osteotomy and internal fixation by double-angle blade plate in patients below 60 years of age with displaced intracapsular fractures and whose time since injury is equal to or more than 3 weeks. Among the 20 fractures which are managed by this procedure, 18 fractures united. We recommend this procedure for the treatment of neglected displaced intracapsular fractures in young patients. PMID- 15826624 TI - Treatment of intertrochanteric fractures in geriatric patients with a modified external fixator. AB - Forty-two geriatric patients who had an intertrochanteric fracture were treated with a semicircular modification of the Ilizarov frame designed by Cattaneo and Catagni between January 1997 and September 2001. Twenty-five of the patients were female, 17 male. The average age of the patients was 77.5 years (range, 63-99). No intraoperative complication occurred. Deep pin-track infection was found in four patients and varus deformity was observed in two patients and shortening of less than 2 cm in 10 patients. Fixator removal was achieved in a mean time of 12 weeks (range, 10-18). No implant failure, refracture or stiffness of knee and hip joint movements was recorded. We concluded that the treatment of intertrochanteric fractures of the elderly patients with our modification provides significant advantages such as minimal operative and anaesthetic risks, no blood loss, early weight-bearing, short hospitalisation time and rapid union time. PMID- 15826625 TI - Home and leisure activities and childhood knee injuries. AB - AIM: To assess the relative occurrence of non motor-vehicle knee injuries and identify important clusters that can be targeted for preventive interventions. METHODS: The study subjects covered 2167 children (0-14 years) who suffered non motor-vehicle knee injuries out of 66870 registered during a three-year period in an established Emergency Department Injury Surveillance System (EDISS). A more serious joint injury was identified in 263 (12%) children, whereas the remaining 1904 children had only soft tissue knee injuries. RESULTS: The incidence of non motor-vehicle knee injuries was estimated at 6.5 per 1000 children-years. Both the incidence of knee injuries and the male-to-female ratio increase with increasing age, reflecting the gender and age pattern of physical activity. Three clusters were identified: The first consisted of more serious knee injuries among older children, frequently resulting after a fall from stairs or a collision in school during winter months; the second cluster consisted of rather minor knee injuries occurring mostly among younger girls at home or in playgrounds, following a fall after stumbling or hit by an object while playing, especially during the summer; the third cluster comprised injuries among older boys, sustained mainly subsequent to overexertion in a sports area. CONCLUSION: Knee injuries tend to be more common among boys but more serious among girls. More and less serious knee injuries tend to fall into distinct clusters that could facilitate prioritization of preventive measures. PMID- 15826626 TI - Peroneal nerve damage associated with the proximal locking screws of the AIM tibial nail. AB - We report a case of division of the deep peroneal nerve resulting from a drill used in the insertion of an oblique proximal locking screw in an AIM tibial intramedullary nail (DePuy). Operative findings and anatomical study indicate there is a risk of damage to the peroneal nerve associated with the oblique proximal locking screws used in this nail design. If a patient has peroneal nerve palsy after nailing of the tibia, the possibility of nerve division should be considered, so that early exploration and repair of the nerve can be performed. PMID- 15826628 TI - Early versus delayed surgical treatment of open tibial fractures: effect on the rates of infection and need of secondary surgical procedures to promote bone union. AB - The purpose of this study was to evaluate whether there was any significant difference in the rates of infection and of secondary surgical procedures to promote bone union, between early(6 h or less after arrival to hospital) and delayed(more than 6 h) surgical treatment of open tibial fractures. Three hundred and eighty-three open tibial fractures were evaluated. 184 fractures had early and 199 had delayed surgical treatment. The rates of infection and secondary surgical procedures to promote bone union of the two groups were compared with univariate and multivariate statistical methods. There was no statistically significant difference between early and delayed treatment groups with respect to overall infection (53/184 versus 51/199 infection rates, P = 0.96), deep infection (8/184 versus 8/199 infection rates, P = 1.0), and rates of secondary surgical procedures to promote bone union (24/184 versus 20/198, P = 0.77). We were unable to demonstrate any significant difference in infection rates or need of secondary procedures to promote bone union, between early and delayed surgical treatment of open tibial fractures. PMID- 15826629 TI - Ilizarov external fixator: acute shortening and lengthening versus bone transport in the management of tibial non-unions. AB - Eighteen patients with tibial shaft non-unions were treated by the Ilizarov method between March 1995 and September 2001 by the senior author. Three subgroups of six patients each were treated by either acute shortening and lengthening, bone transport or simple stabilisation with a frame. All aspects of non-union, infection, shortening, deformity and bone loss were addressed by using Ilizarov principles. There were 10 cases of infected non-unions in the entire series. Bone resection in the shortening group was between 3 and 6 cm (median 4.6) compared to 3-7.5 cm (median 5.9) in the bone transport group. Union was achieved in all the patients with the average time to union at 12.1 months, 17.2 months and 8.0 months, respectively. The bone transport group required additional bone grafting in five patients (83.3%) prior to union compared to one (16.7%) in the acute shortening group. PMID- 15826630 TI - Interlocking nails for displaced metaphyseal fractures of the distal tibia. AB - Metaphyseal fractures of the distal tibia near the ankle joint are difficult to manage. Poor soft tissue coverage and comminution of the fracture complicate open reduction. This prospective study aims to evaluate the practicability of using interlocking nails to treat such fractures. Using the method of closed reduction and internal fixation with a shortened tibial interlocking nail, 20 consecutive cases of distal tibial metaphyseal fractures within 4 cm of the ankle joint line were enrolled from 1997 to 2001. All patients received regular post-operative radiographic check-up and the ankle function was evaluated with the Iowa Ankle Evaluation rating system. Our result was satisfactory and all of the fractures united solidly with a mean union time of 17.2 weeks. No major complication occurred. We conclude that tibial interlocking nailing is a reliable and safe method for managing metaphyseal fractures of the distal tibia near the ankle joint. PMID- 15826631 TI - Arthroscopic assessment of the syndesmosis following ankle fracture. AB - A Maisonneuve ankle fracture without convincing evidence of syndesmotic injury on plain radiographs is described. Stress views and computerised tomography (CT) were also negative. A high index of suspicion led to ankle arthroscopy, which revealed a diastasis, and also an osteochondral fragment. This was treated with two percutaneous diastasis screws and removal of the osteochondral fragment. This case suggests that ankle arthroscopy should be considered as part of the management of syndesmotic injury where conventional imaging techniques fail to show syndesmotic disruption. PMID- 15826632 TI - Inadvertent guide wire advancement in hip fracture fixation with fatal outcome. PMID- 15826633 TI - Clavicular hook-plate: complications of retaining the implant. PMID- 15826635 TI - Premature biomechanical failure of the distal fixation screws of the IC Nail- case report. PMID- 15826637 TI - Effects of hypertonic dextran in hypovolaemic shock: a prospective clinical trial by Alpar EK, Killamapalli VV. PMID- 15826638 TI - A fail safe method of digital tourniquet. PMID- 15826640 TI - Parasite genomes. AB - The availability of genome sequences and the associated transcriptome and proteome mapping projects has revolutionised research in the field of parasitology. As more parasite species are sequenced, comparative and phylogenetic comparisons are improving the quality of gene prediction and annotation. Genome sequences of parasites are also providing important data sets for understanding parasite biology and identifying new vaccine candidates and drug targets. We review some of the preliminary conclusions from examination of parasite genome sequences and discuss some of the bioinformatics approaches taken in this analysis. PMID- 15826641 TI - Integration of tools and resources for display and analysis of genomic data for protozoan parasites. AB - Centralisation of tools for analysis of genomic data is paramount in ensuring that research is always carried out on the latest currently available data. As such, World Wide Web sites providing a range of online analyses and displays of data can play a crucial role in guaranteeing consistency of in silico work. In this respect, the protozoan parasite research community is served by several resources, either focussing on data and tools for one species or taking a broader view and providing tools for analysis of data from many species, thereby facilitating comparative studies. In this paper, we give a broad overview of the online resources available. We then focus on the GeneDB project, detailing the features and tools currently available through it. Finally, we discuss data curation and its importance in keeping genomic data 'relevant' to the research community. PMID- 15826642 TI - Protozoan genomes: gene identification and annotation. AB - The draft sequence of several complete protozoan genomes is now available and genome projects are ongoing for a number of other species. Different strategies are being implemented to identify and annotate protein coding and RNA genes in these genomes, as well as study their genomic architecture. Since the genomes vary greatly in size, GC-content, nucleotide composition, and degree of repetitiveness, genome structure is often a factor in choosing the methodology utilised for annotation. In addition, the approach taken is dictated, to a greater or lesser extent, by the particular reasons for carrying out genome-wide analyses and the level of funding available for projects. Nevertheless, these projects have provided a plethora of material that will aid in understanding the biology and evolution of these parasites, as well as identifying new targets that can be used to design urgently required drug treatments for the diseases they cause. PMID- 15826643 TI - Characterisation of the two most abundant genes in the Haemonchus contortus expressed sequence tag dataset. AB - Analysis of the Haemonchus contortus Expressed sequence tag (EST) dataset revealed that almost 10% of all ESTs (1719 ESTs) belong to a family of related genes. Close analysis of the ESTs suggests that these represent two genes (called here Hc-nim-1 and Hc-nim-2) with multiple alleles of each. These genes show significant similarity to two genes from Caenorhabditis elegans, F54D5.3 (Wormbase accession WBGene00010049, corresponding protein WP:CE28033) and F54D5.4 (WBGene00010050, WP:CE03409) of unknown function. Reverse transcriptase coupled PCR showed that both genes are transcribed from the L4 stage onwards and are transcribed in both male and female adult worms. A partial bacterial recombinant of the Hc-NIM-1 protein was made and used to raise antiserum in rabbits which recognised a 19 kDa antigen in the water soluble protein fraction of adult worms. By immunohistochemistry, the Hc-NIM-1 protein was localised in the hypodermis of the pharyngeal region of adult worms but not posterior in the hypodermis surrounding the reproductive tract. To investigate the function of this novel protein family we conducted a RNA interference experiment for the homologuous proteins in C. elegans. No visible phenotype was detected after simultaneous RNAi treatment for both Ce-F54D5.3 and Ce-F54D5.4. PMID- 15826644 TI - A comparison of match-only algorithms for the analysis of Plasmodium falciparum oligonucleotide arrays. AB - This study is motivated by two data sets which employ a custom Plasmodium falciparum version of the Affymetrix GeneChip, containing only perfect match (PM) oligonucleotides. A PM-only chip cannot be analysed using the standard Affymetrix supplied software. We compared the performance of three match-only algorithms on these data: the Match Only Integral Distribution (MOID) algorithm, Robust Multichip Analysis (RMA), and the Model Based Expression Index (MBEI). We validated the differential expression of several genes using quantitative reverse transcriptase-PCR. We also performed a comparison using two publicly available 'benchmarking' data sets: the Latin Square spike-in data set generated by Affymetrix, and the Gene Logic dilution series. Since we know what the true fold changes are in these special data sets, they are helpful for assessment of expression algorithms. PMID- 15826645 TI - Transcriptional profiling of Entamoeba histolytica trophozoites. AB - We have developed an Entamoeba histolytica genomic DNA microarray and used it to develop a transcriptional profile of 1,971 E. histolytica (HM-1:IMSS) genes. The arrays accurately detected message abundance and 31-47% of amebic genes were expressed under standard tissue culture conditions (levels detectable by Northern blot analysis or RT-PCR respectively). Genes expressed at high levels ( approximately 2% of total) included actin (8.m00351), and ribosomal genes (20.m00312). Moderately expressed genes ( approximately 14% of total) included cysteine proteinase (191.m00117), profilin (156.m00098), and an Argonaute family member (11.m00378). Genes with low-level expression ( approximately 15% of total) included Ariel1 (160.m00087). Genes with very low expression ( approximately 16% of total) and those not expressed ( approximately 52% of total) included encystation-specific genes such as Jacob cyst wall glycoprotein (33.m00261), chitin synthase (3.m00544), and chitinase (22.m00311). Transcriptional modulation could be detected using the arrays with 17% of genes upregulated at least two fold in response to heat shock. These included heat shock proteins (119.m00119 and 279.m00091), cyst wall glycoprotein Jacob (33.m00261), and ubiquitin associated proteins (16.m00343; 195.m00092). Using Caco-2 cells to model the host parasite interaction, we verified that host cell killing was dependent on live ameba. However, surprisingly these events did not appear to induce major transcriptional changes in the parasites. PMID- 15826646 TI - Analysing proteomic data. AB - The rapid growth of proteomics has been made possible by the development of reproducible 2D gels and biological mass spectrometry. However, despite technical improvements 2D gels are still less than perfectly reproducible and gels have to be aligned so spots for identical proteins appear in the same place. Gels can be warped by a variety of techniques to make them concordant. When gels are manipulated to improve registration, information is lost, so direct methods for gel registration which make use of all available data for spot matching are preferable to indirect ones. In order to identify proteins from gel spots a property or combination of properties that are unique to that protein are required. These can then be used to search databases for possible matches. Molecular mass, pI, amino acid composition and short sequence tags can all be used in database searches. Currently the method of choice for protein identification is mass spectrometry. Proteins are eluted from the gels and cleaved with specific endoproteases to produce a series of peptides of different molecular mass. In peptide mass fingerprinting, the peptide profile of the unknown protein is compared with theoretical peptide libraries generated from sequences in the different databases. Tandem mass spectroscopy (MS/MS) generates short amino acid sequence tags for the individual peptides. These partial sequences combined with the original peptide masses are then used for database searching, greatly improving specificity. Increasingly protein identification from MS/MS data is being fully or partially automated. When working with organisms, which do not have sequenced genomes (the case with most helminths), protein identification by database searching becomes problematical. A number of approaches to cross species protein identification have been suggested, but if the organism being studied is only distantly related to any organism with a sequenced genome then the likelihood of protein identification remains small. The dynamic nature of the proteome means that there really is no such thing as a single representative proteome and a complete set of metadata (data about the data) is going to be required if the full potential of database mining is to be realised in the future. PMID- 15826647 TI - Multidrug resistance in parasites: ABC transporters, P-glycoproteins and molecular modelling. AB - Parasitic diseases, caused by protozoa, helminths and arthropods, rank among the most important problems in human and veterinary medicine, and in agriculture, leading to debilitating sicknesses and loss of life. In the absence of vaccines and with the general failure of vector eradication programs, drugs are the main line of defence, but the newest drugs are being tracked by the emergence of resistance in parasites, sharing ominous parallels with multidrug resistance in bacterial pathogens. Any of a number of mechanisms will elicit a drug resistance phenotype in parasites, including: active efflux, reduced uptake, target modification, drug modification, drug sequestration, by-pass shunting, or substrate competition. The role of ABC transporters in parasitic multidrug resistance mechanisms is being subjected to more scrutiny, due in part to the established roles of certain ABC transporters in human diseases, and also to an increasing portfolio of ABC transporters from parasite genome sequencing projects. For example, over 100 ABC transporters have been identified in the Escherichia coli genome, but to date only about 65 in all parasitic genomes. Long established laboratory investigations are now being assisted by molecular biology, bioinformatics, and computational modelling, and it is in these areas that the role of ABC transporters in parasitic multidrug resistance mechanisms may be defined and put in perspective with that of other proteins. We discuss ABC transporters in parasites, and conclude with an example of molecular modelling that identifies a new interaction between the structural domains of a parasite P glycoprotein. PMID- 15826648 TI - Networks in phylogenetic analysis: new tools for population biology. AB - Phylogenetic analysis has changed greatly in the past decade, including the more widespread appreciation of the idea that evolutionary histories are not always tree-like, and may, thus, be best represented as reticulated networks rather than as strictly dichotomous trees. Reconstructing such histories in the absence of a bifurcating speciation process is even more difficult than the usual procedure, and a range of alternative strategies have been developed. There seem to be two basic uses for a network model of evolution: the display of real but unobservable evolutionary events (i.e. a hypothesis of the true phylogenetic history), and the display of character conflict within the data itself (i.e. a summary of the data). These two general approaches are briefly reviewed here, and the strengths and weaknesses of the different implementations are compared and contrasted. Each network methodology seems to have limitations in terms of how it responds to increasing complexity (e.g. conflict) in the data, and therefore each is likely to be more appropriate for one of the two uses than for the other. Several examples using parasitological data sets illustrate the uses of networks within the context of population biology. PMID- 15826649 TI - Protein folding rates estimated from contact predictions. AB - Folding rates of small single-domain proteins that fold through simple two-state kinetics can be estimated from details of the three-dimensional protein structure. Previously, predictions of secondary structure had been exploited to predict folding rates from sequence. Here, we estimate two-state folding rates from predictions of internal residue-residue contacts in proteins of unknown structure. Our estimate is based on the correlation between the folding rate and the number of predicted long-range contacts normalized by the square of the protein length. It is well known that long-range order derived from known structures correlates with folding rates. The surprise was that estimates based on very noisy contact predictions were almost as accurate as the estimates based on known contacts. On average, our estimates were similar to those previously published from secondary structure predictions. The combination of these methods that exploit different sources of information improved performance. It appeared that the combined method reliably distinguished fast from slow two-state folders. PMID- 15826650 TI - Breaking the stereo barrier of amino acid attachment to tRNA by a single nucleotide. AB - Aminoacyl-tRNA synthetases are responsible for attaching amino acid residues to the tRNA 3'-end. The two classes of synthetases approach tRNA as mirror images, with opposite but symmetrical stereochemistries that allow the class I enzymes to attach amino acid residues to the 2'-hydroxyl group of the terminal ribose, whereas, the class II enzymes attach amino acid residues to the 3'-hydroxyl group. However, we show here that the attachment of cysteine to tRNA(Cys) by the class I cysteinyl-tRNA synthetase (CysRS) is flexible; the enzyme is capable of using either the 2' or 3'-hydroxyl group as the attachment site. The molecular basis for this flexibility was investigated. Introduction of the nucleotide U73 of tRNA(Cys) into tRNA(Val) was found to confer the flexibility. While valylation of the wild-type tRNA(Val) by the class I ValRS was strictly dependent on the terminal 2'-hydroxyl group, that of the U73 mutant of tRNA(Val) occurred at either the 2' or 3'-hydroxyl group. Thus, the single nucleotide U73 of tRNA has the ability to break the stereo barrier of amino acid attachment to tRNA, by mobilizing the 2' and 3'-hydroxyl groups of A76 in flexible geometry with respect to the tRNA acceptor stem. PMID- 15826651 TI - Dihydroorotase from Escherichia coli: loop movement and cooperativity between subunits. AB - Escherichia coli dihydroorotase has been crystallized in the presence of the product, L-dihydroorotate (L-DHO), and the structure refined at 1.9A resolution. The structure confirms that previously reported (PDB entry 1J79), crystallized in the presence of the substrate N-carbamyl-D,L-aspartate (D, L-CA-asp), which had a dimer in the asymmetric unit, with one subunit having the substrate, L-CA-asp bound at the active site and the other having L-DHO. Importantly, no explanation for the unusual structure was given. Our results now show that a loop comprised of residues 105-115 has different conformations in the two subunits. In the case of the L-CA-asp-bound subunit, this loop reaches in toward the active site and makes hydrogen-bonding contact with the bound substrate molecule. For the L-DHO bound subunit, the loop faces in the opposite direction and forms part of the surface of the protein. Analysis of the kinetics for conversion of L-DHO to L-CA asp at low concentrations of L-DHO shows positive cooperativity with a Hill coefficient n=1.57(+/-0.13). Communication between subunits in the dimer may occur via cooperative conformational changes of the side-chains of a tripeptide from each subunit: Arg256-His257-Arg258, near the subunit interface. PMID- 15826652 TI - The crystal structure of a novel, latent dihydroorotase from Aquifex aeolicus at 1.7A resolution. AB - Dihydroorotases (EC 3.5.2.3) catalyze the reversible cyclization of carbamoyl aspartate to form dihydroorotate in de novo pyrimidine biosynthesis. The X-ray structures of Aquifex aeolicus dihydroorotase in two space groups, C222(1) and C2, were determined at a resolution of 1.7A. These are the first structures of a type I dihydroorotase, a class of molecules that includes the dihydroorotase domain of mammalian CAD. The type I enzymes are more ancient and larger, at 45 kDa, than the type II enzymes exemplified by the 38 kDa Escherichia coli dihydroorotase. Both dihydroorotases are members of the metallo-dependent hydrolase superfamily, whose members have a distorted "TIM barrel" domain containing the active site. However, A.aeolicus dihydroorotase has a second, composite domain, which the E.coli enzyme lacks and has only one of the two zinc atoms present in the E.coli enzyme. A.aeolicus dihydroorotase is unique in exhibiting significant activity only when complexed with aspartate transcarbamoylase, whereas the E.coli dihydroorotase and the CAD dihydroorotase domain are active as free proteins. The latency of A.aeolicus dihydroorotase can be related to two differences between its structure and that of E.coli dihydroorotase: (1) the monoclinic structure has a novel cysteine ligand to the zinc that blocks the active site and possibly functions as a "cysteine switch"; and (2) active site residues that bind the substrate in E.coli dihydroorotase are located in disordered loops in both crystal structures of A.aeolicus dihydroorotase and may function as a disorder-to-order "entropy switch". PMID- 15826653 TI - LINE-1 retrotransposition requires the nucleic acid chaperone activity of the ORF1 protein. AB - LINE-1 is a highly successful, non-LTR retrotransposon that has played a leading role in shaping mammalian genomes. These elements move autonomously through an RNA intermediate using target-primed reverse transcription (TPRT). L1 encodes two essential polypeptides for retrotransposition, the products of its two open reading frames, ORF1 and ORF2. The exact function of the ORF1 protein (ORF1p) in L1 retrotransposition is unknown, although it is an RNA-binding protein that can act as a nucleic acid chaperone. Here, we investigate the requirements for these two activities in L1 retrotransposition by examining the consequences of mutating two adjacent and highly conserved arginine residues in the ORF1p from mouse L1. Substitution of both arginine residues with alanine strongly reduces the affinity of the protein for single-stranded nucleic acid, whereas substitution of one or both with lysine has only minimal effects on this feature. Rather, the lysine substitutions alter the delicate balance between the ORF1 protein's melting and reannealing activities, thereby reducing its nucleic acid chaperone activity. These findings establish the importance of the nucleic acid chaperone activity of ORF1p to successful L1 retrotransposition, and provide insight into the essential properties of nucleic acid chaperones. PMID- 15826654 TI - The archaeon Haloarcula marismortui has few modifications in the central parts of its 23S ribosomal RNA. AB - Post-transcriptional modifications were mapped in domains II, IV and V of 23S RNA from the archaeon Haloarcula marismortui. The RNA was investigated by two primer extension techniques using reverse transcriptase and three mass spectrometry techniques. One primer extension technique utilized decreasing concentrations of deoxynucleotide triphosphates to detect 2'-O-ribose methylations and other polymerase blocking modifications. In the other, the rRNA was chemically modified, followed by mild alkaline hydrolysis to map pseudo-uridine groups (Psis). RNA fragments for mass spectrometry were isolated from 23S rRNA by site directed RNase H or mung bean nuclease digestion followed by gel purification. Modified RNase digestion fragments were identified with matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) and the modifications were further studied by tandem MS. Psis suggested by the primer extension technique were verified by specific cyanoethylation and mass spectrometric detection. A total of only five post-transcriptionally methylated nucleotides and three Psis were detected in the three 23S rRNA domains. One of the methylated nucleotides has not been reported while a dispute about the number of Psis is solved. The limited number of modified nucleotides suggests that H. marismortui does not have special needs for extensive rRNA modifications. We have performed detailed investigations on the three-dimensional location and molecular interactions of the modified nucleotides by computer analysis. Our results show that all the modified positions are at regions with RNA-RNA contacts and all except one are at the surface of the subunit and in functionally important regions. PMID- 15826655 TI - Human, Drosophila, and C.elegans TDP43: nucleic acid binding properties and splicing regulatory function. AB - TAR DNA binding protein (TDP43), a highly conserved heterogeneous nuclear ribonucleoprotein, was found to down-regulate splicing of the exon 9 cystic fibrosis transmembrane conductance regulator (CFTR) through specific binding to a UG-rich polymorphic region upstream of the 3' splice site. Despite the emergence of new information regarding the protein's nuclear localization and splicing regulatory activity, TDP43's role in cells remains elusive. To investigate the function of human TDP43 and its homologues, we cloned and characterized the proteins from Drosophila melanogaster and Caenorhabditis elegans. The proteins from human, fly, and worm show striking similarities in their nucleic acid binding specificity. We found that residues at two different positions, which show a strong conservation among TDP43 family members, are linked to the tight recognition of the target sequence. Our three-dimensional model of TDP43 in complex with a (UG)(m) sequence predicts that these residues make amino acid side chain to base contacts. Moreover, our results suggest that Drosophila TDP43 is comparable to human TDP43 in regulating exon splicing. On the other hand, C.elegans TDP43 has no effect on exon recognition. TDP43 from C.elegans lacks the glycine-rich domain found at the carboxy terminus of the other two homologues. Mutants of human and fly TDP43 devoid of the C-terminal domain are likewise unable to affect splicing. Our studies suggest that the glycine-rich domain is essential for splicing regulation by human and fly TDP43. PMID- 15826656 TI - Comparative binding of p53 to its promoter and DNA recognition elements. AB - Tumor suppressor p53 is a transcription factor that transactivates a wide range of genes, including those in DNA repair, cell cycle arrest, apoptosis and its own degradation. To estimate the role of selectivity in binding to its promoters, we measured the binding affinities of a tetrameric p53 construct (p53CT) in vitro with 20 of its recognition elements from a variety of representative genes. The binding of full length p53 to four representative sequences exactly paralleled the affinities to p53CT. The binding of p53 to different recognition elements was co-operative and the affinities varied by up to 50-fold. p53 bound with high affinity to the recognition elements of all the genes involved in cell cycle arrest and some of the genes in apoptosis. All of the lower affinity-binding sites were in genes involved in apoptosis. Our quantitative-binding data were in agreement with published cell-based assays. The regulation of p53 activity is in part determined through the specificity of its DNA-binding interactions. PMID- 15826657 TI - Structure of Pyrococcus horikoshii NikR: nickel sensing and implications for the regulation of DNA recognition. AB - The Pyrococcus horikoshii OT3 genome contains a gene (PH0601 or nikR) encoding a protein (PhNikR) that shares 33.8% amino acid sequence identity with Escherichia coli nickel responsive repressor NikR (EcNikR), including many residues that are functionally important in the E.coli ortholog. We succeeded in crystallization and structural characterization of PhNikR in the apo form and two nickel bound forms that exhibit different conformations, open and closed. Moreover, we have identified a putative "low-affinity" nickel-binding pocket in the closed form. This binding site has unusual nickel coordination and exhibits high sensitivity to phosphate in the crystal structure. Analysis of the PhNikR structures and structure-based mutational studies with EcNikR reveals a plausible mechanism of nickel-dependent promoter recognition by the NikR family of proteins. PMID- 15826658 TI - Specific binding of poly(ADP-ribose) polymerase-1 to cruciform hairpins. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) participates in DNA cleavage and rejoining dependent reactions, such as DNA replication, recombination and repair. PARP-1 is also important in transcriptional regulation, although the determinants for its binding to undamaged genomic DNA have not been defined. Previously, we have shown by low-resolution mapping that PARP-1 may bind to the cruciform-forming regions of its own promoter. Here, using DNase I and nuclease P(1) footprinting and atomic force microscopy, we show that PARP-1 binds to stem/loop boundaries of cruciform hairpins. Cleavage of the cruciform by the junction resolvase T4 endonuclease VII is independent of PARP-1, which indicates that PARP-1 does not bind to the four-arm junctions of the cruciform. Thus, PARP-1 differs from other cruciform-binding proteins by binding to hairpin tips rather than to junctions. Furthermore, our data indicate that PARP-1 can interact with the gene regulatory sequences by binding to the promoter-localized cruciforms. PMID- 15826659 TI - In vitro DNA packaging of PRD1: a common mechanism for internal-membrane viruses. AB - PRD1 is the type virus of the Tectiviridae family. Its linear double-stranded DNA genome has covalently attached terminal proteins and is surrounded by a membrane, which is further enclosed within an icosahedral protein capsid. Similar to tailed bacteriophages, PRD1 packages its DNA into a preformed procapsid. The PRD1 putative packaging ATPase P9 is a structural protein located at a unique vertex of the capsid. An in vitro system for packaging DNA into preformed empty procapsids was developed. The system uses cell extracts of overexpressed P9 protein and empty procapsids from a P9-deficient mutant virus infection and PRD1 DNA containing a LacZalpha-insert. The in vitro packaged virions produce distinctly blue plaques when plated on a suitable host. This is the first time that a viral genome is packaged in vitro into a membrane vesicle. Comparison of PRD1 P9 with putative packaging ATPase sequences from bacterial, archaeal and eukaryotic viruses revealed a new packaging ATPase-specific motif. Surprisingly the viruses having this packaging ATPase motif, and thus considered to be related, were the same as those recently grouped together using the coat protein fold and virion architecture. Our finding here strongly supports the idea that all these viruses infecting hosts in all domains of life had a common ancestor. PMID- 15826660 TI - Site-specific DNA-nicking mutants of the heterodimeric restriction endonuclease R.BbvCI. AB - The restriction enzyme R.BbvCI cleaves duplex DNA within a seven base-pair asymmetric recognition sequence, thus: CCTCAGC/GCTGAGG-->CC--TCAGC/GC--TGAGG. We show that R.BbvCI comprises two different subunits, R(1) and R(2); that each subunit contains a catalytic site for DNA strand hydrolysis; and that these sites act independently and strand-specifically. In turn, each catalytic site was inactivated by mutagenesis to form dimeric enzymes in which only one site remained functional. The altered enzymes hydrolyzed just one strand of the recognition sequence, nicking the DNA rather than cleaving it. Enzymes in which the catalytic site in the R(1) subunit remained functional nicked the bottom strand of the sequence, producing CCTCAGC/GC--TGAGG, while those in which the catalytic site in the R(2) subunit remained functional nicked the top strand, producing CC--TCAGC/GCTGAGG. These DNA-nicking enzymes could prove useful for investigation of DNA repair, recombination, and replication, and for laboratory procedures that initiate from nicks, such as DNA degradation, synthesis, and amplification. PMID- 15826661 TI - Cleavage of individual DNA strands by the different subunits of the heterodimeric restriction endonuclease BbvCI. AB - BbvCI cleaves an asymmetric DNA sequence, 5'-CC downward arrow TCAGC-3'/5'-GC downward arrow TGAGG-3', as indicated. While many Type II restriction enzymes consist of identical subunits, BbvCI has two different subunits: R(1), which acts at GC downward arrow TGAGG; and R(2), which acts at CC downward arrow TCAGC. Some mutants of BbvCI with defects in one subunit, either R(1)(-)R(2)(+) or R(1)(+)R(2)(-), cleave only one strand, that attacked by the native subunit. In analytical ultracentrifugation at various concentrations of protein, wild-type and mutant BbvCI enzymes aggregated extensively, but are R(1)R(2) heterodimers at the concentrations used in DNA cleavage reactions. On a plasmid with one recognition site, wild-type BbvCI cleaved both strands before dissociating from the DNA, while the R(1)(-)R(2)(+) and R(1)(+)R(2)(-) mutants acted almost exclusively on their specified strands, albeit at relatively slow rates. During the wild-type reaction, the DNA is cleaved initially in one strand, mainly that targeted by the R(1) subunit. The other strand is then cleaved slowly by R(2) before the enzyme dissociates from the DNA. Hence, the nicked form accumulates as a transient intermediate. This behaviour differs from that of many other restriction enzymes, which cut both strands at equal rates. However, the activities of the R(1)(+) and R(2)(+) subunits in the wild-type enzyme can differ from their activities in the R(1)(+)R(2)(-) and R(1)(-)R(2)(+) mutants. Each active site in BbvCI therefore influences the other. PMID- 15826662 TI - The three-dimensional structure of the bifunctional 6-hydroxymethyl-7,8 dihydropterin pyrophosphokinase/dihydropteroate synthase of Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae and other fungi, the enzymes dihydroneopterin aldolase, 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) and dihydropteroate synthase (DHPS) are encoded by a polycistronic gene that is translated into a single polypeptide having all three functions. These enzymatic functions are essential to both prokaryotes and lower eukaryotes, and catalyse sequential reactions in folate biosynthesis. Deletion or disruption of either function leads to cell death. These enzymes are absent from mammals and thus make ideal antimicrobial targets. DHPS is currently the target of antifolate therapy for a number of infectious diseases, and its activity is inhibited by sulfonamides and sulfones. These drugs are typically used as part of a synergistic cocktail with the 2,4-diaminopyrimidines that inhibit dihydrofolate reductase. A gene encoding the S.cerevisiae HPPK and DHPS enzymes has been cloned and expressed in Escherichia coli. A complex of the purified bifunctional polypeptide with a pterin monophosphate substrate analogue has been crystallized, and its structure solved by molecular replacement and refined to 2.3A resolution. The polypeptide consists of two structural domains, each of which closely resembles its respective monofunctional bacterial HPPK and DHPS counterpart. The mode of ligand binding is similar to that observed in the bacterial enzymes. The association between the domains within the polypeptide as well as the quaternary association of the polypeptide via its constituent DHPS domains provide insight into the assembly of the trifunctional enzyme in S.cerevisiae and probably other fungal species. PMID- 15826663 TI - Crystal structure of earthworm fibrinolytic enzyme component B: a novel, glycosylated two-chained trypsin. AB - The earthworm fibrinolytic enzyme (EFE), belonging to a group of serine proteases with strong fibrinolytic activity, has been used in a mixture as an oral drug for prevention and treatment of thrombosis in East Asia. The EFE component b (EFE-b) is one of seven EFE components from Eisenia fetida, and among them it has nearly the highest fibrinolytic activity. Here, we report its crystal structure at a resolution of 2.06A. The structural analysis shows that EFE-b should be classified as a trypsin from earthworm. However, it is distinct from other trypsins. It is a two-chained protease with an N-terminal, pyroglutamated light chain and an N-glycosylated heavy chain. Furthermore, the heavy chain contains a novel structural motif, an eight-membered ring resulting from a disulfide bridge between two neighboring cysteine residues, and a cis peptide bond exists between these two cysteine residues. The crystal structure of EFE-b provides the structural basis for its high level of stability and reveals its complicated post translational modifications in earthworm. This structure is the first reported for a glycosylated two-chained trypsin, which may provide useful clues to explain the origin and evolution of the chymotrypsin family. PMID- 15826664 TI - The GB1 amyloid fibril: recruitment of the peripheral beta-strands of the domain swapped dimer into the polymeric interface. AB - Three-dimensional domain swapping has been evoked as a mechanism for oligomerization of proteins. Here, we show for the immunoglobulin-binding domain B1 of streptococcal protein G (GB1) that fibril formation is observed readily for variants that exist as domain-swapped dimers. No fibril was formed by a revertant that exhibits the stable wild-type GB1 fold or a mutant comprising a highly destabilized, fluctuating ensemble of conformers. Structural features of the GB1 amyloid fibril were characterized by cysteine disulfide cross-linking. Residues in the outer edge beta-strands of the domain-swapped dimer readily form intermolecular disulfide bonds prior to and during fibril formation. On the basis of these data, a structural model for the assembly of domain-swapped dimers into a polymeric structure of the GB1 fibril is proposed. PMID- 15826665 TI - A structure-based database of antibody variable domain diversity. AB - The diversity of natural antibodies is limited by the genetic mechanisms that engender diversity and the functional requirements of antigen binding. Using an in vitro-evolved autonomous heavy chain variable domain (V(H)H-RIG), we have investigated the limits of structurally-tolerated diversity in the three complementarity-determining regions and a fourth loop within the third framework region. We determined the X-ray crystal structure of the V(H)H-RIG domain at 1.9A resolution and used it to guide the design of phage-displayed libraries encompassing the four loops. The libraries were subjected to selections for structural stability, and a database of structurally-tolerated sequences was compiled from the sequences of approximately 1000 unique clones. The results reveal that all four loops accommodate significantly greater diversity than is observed in nature. Thus, it appears that most sequence biases in the natural immune repertoire arise from factors other than structural constraints and, consequently, it should be possible to enhance the functions of antibodies significantly through in vitro evolution. PMID- 15826666 TI - Solution structure of the Ran-binding domain 2 of RanBP2 and its interaction with the C terminus of Ran. AB - The termination of export processes from the nucleus to the cytoplasm in higher eukaryotes is mediated by binding of the small GTPase Ran as part of the export complexes to the Ran-binding domains (RanBD) of Ran-binding protein 2 (RanBP2) of the nuclear pore complex. So far, the structures of the first RanBD of RanBP2 and of RanBP1 in complexes with Ran have been known from X-ray crystallographic studies. Here we report the NMR solution structure of the uncomplexed second RanBD of RanBP2. The structure shows a pleckstrin homology (PH) fold featuring two almost orthogonal beta-sheets consisting of three and four strands and an alpha-helix sitting on top. This is in contrast to the RanBD in the crystal structure complexes in which one beta-strand is missing. That is probably due to the binding of the C-terminal alpha-helix of Ran to the RanBD in these complexes. To analyze the interaction between RanBD2 and the C terminus of Ran, NMR titration studies with peptides comprising the six or 28 C-terminal residues of Ran were performed. While the six-residue peptide alone does not bind to RanBD2 in a specific manner, the 28-residue peptide, including the entire C-terminal helix of Ran, binds to RanBD2 in a manner analogous to the crystal structures. By solving the solution structure of the 28mer peptide alone, we confirmed that it adopts a stable alpha-helical structure like in native Ran and therefore serves as a valid model of the Ran C terminus. These results support current models that assume recognition of the transport complexes by the RanBDs through the Ran C terminus that is exposed in these complexes. PMID- 15826667 TI - Structure of S5a bound to monoubiquitin provides a model for polyubiquitin recognition. AB - Ubiquitin is a key regulatory molecule in diverse cellular events. How cells determine the outcome of ubiquitylation remains unclear; however, a likely determinant is the specificity of ubiquitin receptor proteins for polyubiquitin chains of certain length and linkage. Proteasome subunit S5a contains two ubiquitin-interacting motifs (UIMs) through which it recruits ubiquitylated substrates to the proteasome for their degradation. Here, we report the structure of S5a (196-306) alone and complexed with two monoubiquitin molecules. This construct contains the two UIMs of S5a and we reveal their different ubiquitin binding mechanisms and provide a rationale for their unique specificities for different ubiquitin-like domains. Furthermore, we provide direct evidence that S5a (196-306) binds either K63-linked or K48-linked polyubiquitin, and in both cases prefers longer chains. On the basis of these results we present a model for how S5a and other ubiquitin-binding proteins recognize polyubiquitin. PMID- 15826668 TI - Recognizing and defining true Ras binding domains I: biochemical analysis. AB - Common domain databases contain sequence motifs which belong to the ubiquitin fold family and are called Ras binding (RB) and Ras association (RalGDS/AF6 Ras associating) (RA) domains. The name implies that they bind to Ras (or Ras-like) GTP-binding proteins, and a few of them have been documented to qualify as true Ras effectors, defined as binding only to the activated GTP-bound form of Ras. Here we have expressed a large number of these domains and investigated their interaction with Ras, Rap and M-Ras. While their (albeit weak) sequence homology suggest that the domains adopt a common fold, not all of them bind to Ras proteins, irrespective of whether they are called RB or RA domains. We used fluorescence spectroscopy and isothermal titration calorimetry to show that the binding affinities vary over a large range, and are usually specific for either Ras or Rap. Moreover, the specificity is dictated by a set of key residues in the interface. Stopped-flow kinetic analysis showed that the association rate constants determine the different affinities of effector binding, while the dissociation rate constants are in a similar range. Manual sequence analysis allowed us to define positively charged sequence epitopes in certain secondary structure elements of the ubiquitin fold (beta1, beta2 and alpha1) which are located at similar positions and comprise the hot spots of the binding interface. These residues are important to qualify an RA/RB domain as a true candidate Ras or Rap effector. PMID- 15826669 TI - Recognizing and defining true Ras binding domains II: in silico prediction based on homology modelling and energy calculations. AB - Considering the large number of putative Ras effector proteins, it is highly desirable to develop computational methods to be able to identify true Ras binding molecules. Based on a limited sequence homology among members of the Ras association (RA) and Ras binding (RB) sub-domain families of the ubiquitin super family, we have built structural homology models of Ras proteins in complex with different RA and RB domains, using the FOLD-X software. A critical step in our approach is to use different templates of Ras complexes, in order to account for the structural variation among the RA and RB domains. The homology models are validated by predicting the effect of mutating hot spot residues in the interface, and residues important for the specificity of interaction with different Ras proteins. The FOLD-X calculated energies of the best-modelled complexes are in good agreement with previously published experimental data and with new data reported here. Based on these results, we can establish energy thresholds above, or below which, we can predict with 96% confidence that a RA/RB domain will or will not interact with Ras. This study shows the importance of in depth structural analysis, high quality force-fields and modelling for correct prediction. Our work opens the possibility of genome-wide prediction for this protein family and for others, where there is enough structural information. PMID- 15826670 TI - X-ray structure of a water-soluble analog of the membrane protein phospholamban: sequence determinants defining the topology of tetrameric and pentameric coiled coils. AB - Phospholamban (PLB) is a pentameric transmembrane protein that regulates the Ca(2+)-dependent ATPase SERCA2a in sarcoplasmic reticulum membranes. We previously described the computational design of a water-soluble variant of phospholamban, WSPLB, which reproduced many of the structural and functional properties of the native membrane-soluble protein. While the full-length WSPLB forms a pentamer in solution, a truncated variant forms very stable tetramers. To obtain insight into the tetramer-pentamer cytoplasmic switch, we solved the crystal structure of the truncated construct, WSPLB 21-52. This peptide has a heptad sequence repeat with Leu residues at a- and Ile at d-positions from residues 31-52. The crystal structure revealed that WSPLB 21-52 adopted an antiparallel tetrameric coiled coil. This topology contrasts with the parallel topology of an analogue of the coiled-coil of GCN4 with the same Leu(a) Ile(d) repeat. Analysis of these structures revealed how the nature of the partially exposed residues at e- and g-positions influence the topology formed by the bundle. We also constructed a model for the pentameric form of PLB using the coiled-coil parameters derived from a single monomer in the tetrameric structure. This model suggests that both buried and interfacial hydrogen bonds are important for stabilizing the parallel pentamer. PMID- 15826671 TI - Hypothesis: the origin of life in a hydrogel environment. AB - A hypothesis is proposed that the first cell(s) on the Earth assembled in a hydrogel environment. Gel environments are capable of retaining water, oily hydrocarbons, solutes, and gas bubbles, and are capable of carrying out many functions, even in the absence of a membrane. Thus, the gel-like environment may have conferred distinct advantages for the assembly of the first cell(s). PMID- 15826672 TI - Induction and the Turing-Child field in development. AB - The central problem in biological development is the understanding of epigenesis. The dominant theory of development in the last 80 years that also purports to explain epigenesis is induction theory. It suggests that development is driven by sequential inductions where each "induction" (in one sense of the word induction) is effected by the action of an inducing part of the embryo on a responding part of the embryo. The theory stems from Spemann and Mangold (W.Roux' Arch.f.Entw.d.Organis.u.mikrosk.Anat.100 (1924) 599) who transplanted a tissue from the dorsal blastopore lip of Triturus into the ventral ectoderm of another gastrula and thus initiated and "induced" (in another sense of the word induction) gastrulation and embryogenesis in the ventral side of the host that became a double embryo (siamese twins). We explain this induction, i.e. the formation of the double embryo, according to the Child theory and the Turing Gierer-Meinhardt theory when it is also assumed that cAMP and ATP are the Turing activator and inhibitor, respectively. Spemann and Mangold (W.Roux' Arch.f.Entw.d.Organis.u.mikrosk.Anat.100 (1924) 599) also suggested that the ingressing mesoderm induces the overlying ectoderm to form the neural plate and neural tube. This 'neural induction', the 'primary embryonic induction', became the cornerstone of induction theory, i.e. of the sequential induction concept referred to above. But we argue that the metabolic gradients that precede and accompany neurulation, as obtained by Child, also for Triturus, arise through a Turing self-organization if it is assumed that cAMP and ATP are the Turing morphogens, and these gradients are the cause and primary event of neurulation. Thus there is no need to invoke the 'neural induction'. It is argued that fundamental events such as gastrulation and also organ formation are caused by the Turing-Child field and not by sequential induction. Similar principles, such as bud formation caused by a radial metabolic pattern that transforms to a longitudinal pattern, govern the formation, for example, of the mouth and the gut. The formation and localization of bottle cells is explained according to the Child-Turing field and modern biochemistry. The chemical metabolic pre-pattern precedes, and causes, morphogenesis and differentiation as envisaged by Turing. The Spemann and Mangold (W.Roux' Arch.f.Entw.d.Organis.u.mikrosk.Anat.100 (1924) 599) transplantation experiment when performed on a sea urchin duplicates not only the phenotype but also the metabolic (reduction) pattern. These experimental results, by Horstadius, predicted by Child, follow from the Turing-Gierer Meinhardt theory if it is assumed that cAMP and ATP are the Turing morphogens. If the transplantation is performed not onto the whole sea urchin but onto only a part of it, that manifests only a part of the metabolic pattern, then from the part a phenotypic whole underlain by a normal and a whole metabolic pattern can be rescued. These experimental results of Horstadius follow from Turing theory if cAMP and ATP are the Turing morphogens. Understanding how to transform a part into a whole can be valuable in regenerative medicine. Unspecific induction of a secondary amphibian embryo is similar to the induction of posterior structures at the anterior pole of an insect, and the "double abdomen" (and Kalthoff's experimental results) of the midge Smittia resulting from UV irradiation of the anterior pole, can be explained by Meinhardt theory of unspecific induction if ATP is the Turing morphogen. When not working on regeneration, Child investigated intact living organisms and his observation method was not disruptive to normal development, whereas workers in induction theory work with pieces and in general disrupt normal development. We conclude that the Turing-Child field causes all development and explains epigenesis. Sequential induction does not explain epigenesis and does not exist in normal development. But induction in the sense of a transplantation leading to double embryo or rescuing a whole phenotype from a part is of interest. PMID- 15826673 TI - Modeling embryogenesis and cancer: an approach based on an equilibrium between the autostabilization of stochastic gene expression and the interdependence of cells for proliferation. AB - A large amount of data demonstrating the stochastic nature of gene expression and cell differentiation has accumulated during the last 40 years. These data suggest that a gene in a cell always has a certain probability of being activated at any time and that instead of leading to on and off switches in an all-or-nothing fashion, the concentration of transcriptional regulators increases or decreases this probability. In order to integrate these data in an appropriate theoretical frame, we have tested the relevance of the selective model of cell differentiation by computer simulation experiments. This model is based on stochastic gene expression controlled by cellular interactions. Our results show that it is readily able to produce tissue organization. A model involving only two cells generated a bi-layer cellular structure of finite growth. Cell death was not a drawback but an advantage because it improved the viability of this bi layer structure. However, our results also show that cellular interactions cannot be simply based on raw selection between cells. Instead, tissue coordination includes at least two basic components: phenotypic autostabilization (differentiated cells stabilize their own phenotype) and interdependence for proliferation (differentiated cells stimulate the proliferation of alien phenotypes). In this modified autostabilization-selection model, cellular organization and growth arrest result from a quantitative equilibrium between the parameters controlling these two processes. An imbalance leads to tissue disorganization and invasive cancer-like growth. These findings suggest that cancer does not result solely from mutations in the cancerous cell but from the progressive addition of several small alterations of the equilibrium between autostabilization and interdependence for proliferation. In this frame, it is not solely the cancerous cell that is abnormal. The whole organism is involved. Tumor growth is a local effect of an imbalance between all the factors involved in tissue organization. PMID- 15826674 TI - Fertility in South Australian commercial Merino flocks: sources of reproductive wastage. AB - To identify reasons for low fertility in the Merino in South Australia, we defined the extent and sources of reproductive wastage in flocks of maiden (n=14) and mature age (n=54) Merino ewes managed on 43 properties over 4 year. In a second study, reproductive wastage was examined in a very high ovulating flock of mature age South Australian Merino ewes heterozygous for the FecB gene, mated to lamb in either the autumn or spring. Losses in the latter flock were examined in less detail than the large-scale study, but allowed wastage to be partitioned between pre- and peri/post-natal sources. In the first experiment, reproductive wastage between mating and weaning was estimated at 59.7 potential lambs or ova lost per 100 ewes exposed to rams, representing 42.4% of ova shed. The main source of wastage occurred at lambing (55.6% of total loss), with death of twins a major contributor (35.3% of total loss). Other important sources of wastage were from partial failure of multiple ovulation (PFMO; 20.4% of total loss) and from ewes mating but not lambing (13.2% of total loss). Reproductive wastage did not vary with either age of ewe (maiden versus mature) or season of mating (October-December versus January-March). Mean ovulation rate, estimated as 141 per 100 ewes ovulating (range 100-200), varied with age of ewe (129 versus 144; maiden versus mature) but not with season of mating or year (1990-1991). Values for fertility, fecundity and lamb survival were 90, 127 and 73%, respectively. Fecundity of maiden ewes was lower than that of mature ewes (116 versus 130). Survival of single and twin lambs between birth and tail docking was 83 and 56%, respectively. Net reproductive efficiency, or lambs weaned per 100 ewes exposed to rams, averaged 81 (range 31-122). In the second experiment, mean ovulation rate of FecB Merino ewes was 316 per 100 ewes ovulating, with 242 ova (potential lambs) lost between ovulation and tail docking. This loss was equally shared between pre- and peri/post-natal sources (123 versus 119), and demonstrates severe limitation of the Merino to successfully bear and rear multiple litters. We concluded that future research in commercial Merino flocks be focused on lamb mortality, particularly of twins, and on PFMO, the major source of embryo loss. PMID- 15826675 TI - Use of strontium for bovine oocyte activation. AB - Strontium efficiently activates mouse oocytes, however, there is limited information on its use in cattle. Thus, the objective of this study was to establish a suitable protocol for activating bovine oocyte with strontium. For pronuclear development, the absence of calcium and magnesium in the activation medium (TALP) with 10 and 50 mM strontium (34.4 and 53.1%, respectively) was superior to the complete TALP (6.5 and 19.4%, respectively). In all activation media, better results were observed with 25 and 50 mM strontium (21.9-53.1 and 19.4-53.1%, respectively). Incubation for 4 h promoted similar results in all strontium concentrations. However, strontium at 15, 20, and 25 mM for 6 and 8 h (40.7, 46.7, and 48.3%, and 29.3, 48.3, and 40.7%, respectively) were superior to control (15.5 and 10%, respectively). After in vitro maturation for 26 h, strontium (S; 20 mM in Ca2+- and Mg2+-free TALP for 6 h), ionomycin+strontium (IS), and strontium+ionomycin (SI) (60, 63.3, and 65%, respectively) were similar in pronuclear development and superior to ionomycin (I; 5 microM for 5 min; 36.7%). In treatments S and I, only 1 PN zygotes were observed. In treatment S, most of them had 1 and 2 PB (35.7 and 60.7%, respectively), and in treatment I, 0, 1, and 2 PB (14.3, 57.1, and 28.6%, respectively). Most of the zygotes in treatment IS and SI were 1 PN 2 PB (77.4 and 61.6%, respectively). The number of oocytes with clusters of cortical granules was similar in all treated groups (11 29%). Cortical granule exocytosis in treatment IS (68%) was similar to S (54%) and superior to I, SI, and control (27, 45, and 5.0%, respectively). Cleavage and blastocyst rates were similar for S, I, IS, and SI treatments (61.7-76.7, and 8.3 13.3%, respectively) and the same was observed for ICM, TE, and total cell number, and ICM/total cell ratio (22-25, 64-69, and 86-95, and 0.26-0.27). In conclusion, strontium may be efficiently applied for bovine oocyte activation at 20 mM in Ca2+- and Mg2+-free TALP medium for 6 h. PMID- 15826676 TI - Cryopreservation of channel catfish sperm: effects of cryoprotectant exposure time, cooling rate, thawing conditions, and male-to-male variation. AB - The purpose of this study was to extend previous work on the cryopreservation of channel catfish (Ictalurus punctatus) sperm. The objectives were to compare the effects of freezing and thawing on motility of sperm for: (1) 1 or 48-h exposure before freezing to 5% methanol and use of 0.5 or 0.25 mL straws; (2) 1 h or 5-day exposure before freezing to 5% methanol; (3) cooling at 45 or 3 degrees C/min; (4) thawing at 30, 40 or 50 degrees C using 5 or 10 s duration, and (5) cryopreservation with 5 or 10% methanol of samples from 50 males to analyze male to-male variation. No differences were found in motility reduction for 1 or 48 h exposure times in 5% methanol, for use of 0.5 or 0.25 mL straws, or for 1 h or 5 day exposures in 5% methanol. A cooling rate of 45 degrees C/min resulted in lower motility reduction (33+/-9%) than a rate of 3 degrees C/min (83+/-13%) (P=0.002). A thawing temperature of 50 degrees C resulted in lower motility reduction (25+/-14%) than 30 degrees C (51+/-21%) or 40 degrees C (59+/-11%) (P=0.001). A thawing duration of 10 s resulted in lower motility reduction (38+/ 12%) than a duration of 5 s (52+/-12%) (P=0.005), and there was an interaction between thawing temperature and duration (P=0.050). A concentration of 5% methanol resulted in lower motility reduction (43+/-17%) than 10% methanol (67+/ 14%) (P=0.001). Regression analysis showed no relationship between motility before freezing and after thawing for 5% methanol (r2=0.012) or 10% methanol (r2=0.011). PMID- 15826677 TI - Seasonal changes in fecal testosterone concentrations and their relationship to the reproductive behavior, antler cycle and grouping patterns in free-ranging male Pampas deer (Ozotoceros bezoarticus bezoarticus). AB - The purpose of this study was to validate noninvasive endocrine monitoring techniques for Pampas deer and to evaluate seasonal changes in testicular steroidogenic activity and their correlation to reproductive behavior, antler cycle and group size. Thus, fecal samples, behavioral data and observations of antler status were collected at monthly intervals during 1 year from free-ranging Pampas deer stags (three radio-collared individuals and 15 random individuals) living in Emas National Park, Brazil (18 degrees S latitude). Fecal steroids were extracted using 80% methanol and steroid concentrations were quantified by a commercial enzyme immunoassay (EIA). Fecal testosterone concentrations peaked in December-January (summer), March (early autumn) and in August-September (winter spring), with minimal values from April-July. Reproductive behavior had two peaks, the first in December-January, characterized by predominately anogenital sniffing, flehmen, urine sniffing, chasing and mounting behavior, and the second peak in July-September (behavior primarily related to gland marking). There were significant correlations between fecal testosterone and reproductive behavior (r=0.490), and between fecal testosterone and antler phases (r=0.239). Antler casting and regrowth occurred under low testosterone concentrations, whereas velvet shedding was associated with high concentrations of testosterone. We inferred that Pampas deer stags exhibited a seasonal cycle that modulated sexual behavior and the antler cycle, and we concluded that fecal steroid analysis was a practical and reliable non-invasive method for the evaluation of the endocrine status of free-ranging Pampas deer. PMID- 15826678 TI - Alpha-tocopherol improves biochemical and dynamic parameters in cryopreserved boar semen. AB - Cryopreservation is associated with the production of reactive oxygen species which lead to lipid peroxidation of sperm membranes. The objective was to determine an alpha-tocopherol concentration capable of improving the quality of cryopreserved porcine semen. Boar spermatozoa frozen with 200, 500 or 1000 microg/mL alpha-tocopherol were thawed and incubated at 37 degrees C for 4 h. Routine parameters of semen quality, susceptibility to lipid peroxidation 2 thiobarbituric acid (TBARS) and oxygen uptake were evaluated. Motility was higher (P<0.05) in samples treated with different concentrations of alpha-tocopherol up to 2 h of incubation. Viability and acrosome integrity significantly decreased during incubation (no significant differences between treatments). Two hundred micrograms per milliliter alpha-tocopherol protected spermatozoa against lipid peroxidation during incubation, but 1000 microg/mL failed to protect after 2 h of incubation. There was a negative association between TBARS and motility, suggesting that lipid peroxidation affected sperm motility. Both control and 200 microg/mL alpha-tocopherol samples preserved the capacity to generate oxidative energy up to 1 h of incubation. The addition of 200 microg/mL alpha-tocopherol in the semen extender could be useful to preserve boar spermatozoa against the oxidative stress generated by cryopreservation. PMID- 15826679 TI - Ovarian follicular development and oocyte quality in anestrous ewes treated with melatonin, a controlled internal drug release (CIDR) device and follicle stimulating hormone. AB - The objective of the current study was to determine the effects of hormonal treatments on ovarian follicular development and oocyte quality in anestrous ewes. Multiparous crossbred (RambouilletxTarghee) ewes were given melatonin implants (MEL) and/or controlled internal drug release (CIDR) devices in conjunction with follicle stimulating hormone (FSH) during anestrus (March-May). In Experiment 1, ewes (n=25) were assigned randomly to four groups (n=4-7/group) in a 2x2 factorial arrangement [+/-MEL and +/-CIDR], resulting in Control (no treatment), CIDR, MEL, and MEL/CIDR groups, respectively. Ewes received an implant containing 18 mg of melatonin (Melovine) on Day 42 and/or a CIDR from Days 7 to 2 (Day 0: oocyte collection). In Experiment 2, ewes (n=12) were assigned randomly to two groups (n=6/group; 1CIDR or 2CIDR) and received the same type of melatonin implant on Day 60. All ewes received a CIDR device from Days 22 to -17 and 2CIDR ewes received an additional CIDR device from Days -10 to -2. In both experiments, ewes were given FSH im twice daily (morning and evening) on Days -2 and -1 (Day -2: 5 units/injection; Day -1: 4 units/injection). On the morning of Day 0, ovaries were removed, follicles>or=1 mm were counted, and oocytes were collected. Thereafter oocytes were matured and fertilized in vitro. In Experiment 1, the number of visible follicles and the rates of oocyte recovery and in vitro maturation were similar (P>0.10) for Control, CIDR, MEL and MEL/CIDR (overall 29.7+/-2.9%, 89.9+/-7.1% and 95.0+/-2.0%, respectively). The rates of in vitro fertilization (IVF) were lower (P<0.01) for CIDR and MEL/CIDR than for Control and MEL groups (10.3% and 10.1% versus 20.0% and 18.5%, respectively). In Experiment 2, the number of visible follicles, and the rates of oocyte recovery and in vitro maturation were similar (P>0.10) for 1CIDR and 2CIDR groups (overall 27.3+/-3.2%, 92.1+/-2.7% and 90.2+/-1.9%, respectively). However, the rates of IVF were lower (P<0.01) for 2CIDR than 1CIDR group (30.2% versus 58.0%, respectively). In summary, when treatment with P4 commenced only 2 d before oocyte collection, rates of IVF were reduced in both experiments. Therefore, progestin treatment protocols used in ovine IVF programs should be carefully designed to minimize adverse effects on fertilization rates. In addition, melatonin treatment did not affect follicular development and oocyte quality for anestrous ewes. PMID- 15826680 TI - Apoptosis in cumulus cells, but not in oocytes, may influence bovine embryonic developmental competence. AB - Aim of our study was to clarify if the occurrence of apoptosis in oocytes and cumulus cells is correlated to bovine oocyte developmental competence. The cumulus-oocyte complexes (COCs) were selected according to cumulus status: G1 with more than five layers of compact cumulus cells, G2 with one to five layers of compact cumulus cells and G3 with expanded cumulus cells. The degree of apoptosis in cumulus cells and oocytes measured by caspase staining and TUNEL assay before and after maturation, and 24 h post-insemination was compared to the cleavage, blastocyst formation and hatching rates of each group. Highest cleavage, blastocyst and hatching rates were found in cumulus-oocyte complexes with more than five layers of compact cumulus cells, but no apoptosis was detected in immature or in vitro matured oocytes, regardless of the cumulus status. Many cumulus cells contained active caspases before maturation, but caspase activity declined dramatically after maturation. TUNEL positive cells were rarely observed in each cumulus-oocyte complex upon oocyte recovery, but a huge increase of them was seen after in vitro maturation. Significantly more TUNEL and caspase positive cells were found in G2 cumulus-oocyte complexes. Our results suggest that: (i) oocyte apoptosis does not account for the inferior oocyte quality of G2 and G3; (ii) apoptosis occurs in cumulus cells regardless of the number and compactness of cumulus cells; and (iii) the degree of apoptosis in the compact cumulus-oocyte complexes (G1 and G2) is negatively correlated to the developmental competence of oocyte. PMID- 15826681 TI - FSH and LH plasma levels in bitches with differences in risk for urinary incontinence. AB - To determine whether the height of the plasma gonadotropin levels after spaying is associated with urinary incontinence, the concentrations of plasma follicle stimulating hormone (FSH) and luteinizing hormone (LH) were determined once in 191 intact and 308 spayed bitches. The bitches were grouped according to their risk for urinary incontinence and the medians of their respective gonadotropin levels were compared. For intact anestrous bitches, the FSH- and LH-plasma concentrations were 5.2 (4, 8) ng/mL (median (Q1, Q3)) and 0.5 (0.5-0.5) ng/mL, respectively. In the first year after spaying, the gonadotropin concentrations rose significantly, then stabilised at a level around 10 times those of intact bitches (FSH 62.5 (44, 91) ng/mL; LH 6.1(4, 11) ng/mL). The plasma gonadotropin concentrations of long-term spayed (>12 months) continent bitches (n=209) were higher (FSH 66.8 (46, 104) ng/mL; LH 6.5 (4, 11) ng/mL) than in spayed incontinent bitches (n=60) (FSH 51.5 (38, 74) ng/mL; LH 5.5 (3, 8) ng/mL), the latter also had a higher body weight. Multiple regression analysis showed that the FSH-plasma concentration and not the body weight was decisive for the occurrence of urinary incontinence. The results of this study suggest that levels of gonadotropins are associated, directly or indirectly in the pathophysiology of urinary incontinence after spaying. PMID- 15826682 TI - Interaction between lactation, photoperiodism and male effect in German Merino ewes. AB - A study with 93 German Merino ewes was performed from January until the end of March to clarify the relative importance of lactation, photoperiodism and ram effect on cyclic activity and lambing data. Ovarian activity was registered by progesterone concentrations in blood plasma three times weekly. Half of the ewes were kept under supplemental light (20 h/day) for the last 6 weeks of lactation and additionally 3 weeks post-weaning, the other half were kept under natural photoperiod but were weaned simultaneously. Thereafter, light was reduced to natural photoperiod and rams were introduced to half of the ewes, of both light reduced and photoperiod group. Ewes entered cyclicity during lactation gradually, but at weaning 56% of photoperiod ewes and 53% of supplemental light ewes were still acyclic. After weaning, resumption of cyclic activity before ram introduction was more pronounced (P<0.05) in the photoperiod group (75% cyclic) than in the supplemental light group (51% cyclic). Ram introduction led to cyclicity in all ewes. Light reduction without ram slightly increased cyclicity but 57% were still acyclic. In the photoperiod group without ram no ewe entered cyclicity and two ewes even ceased cycling again. Data show that German Merinos still have a remarkable lactational anoestrus but are extremely sensitive to ram. Light reduction has no direct effect on cyclicity but is likely to contribute to the elevated ovulation rate so that a combination with the ram effect led to a higher lambing rate (1.94) compared to photoperiod and ram (1.55). PMID- 15826683 TI - Bovine blastocyst development rate in vitro is influenced by selection of oocytes by brillant cresyl blue staining before IVM as indicator for glucose-6-phosphate dehydrogenase activity. AB - The aim of this present study was to increase the efficiency of blastocyst production from cows after in vitro maturation/fertilization (IVM/IVF) by oocyte selection before maturation. Oocytes were selected on the basis of brillant cresyl blue (BCB) staining, used to indicate glucose-6-phosphate dehydrogenase (G6PDH) activity. To re-valuate the hypothesis that growing oocytes are expected to have a high level of active G6PDH, while mature oocytes have low G6PDH activity, cumulus oocyte complexes (COCs) were recovered from slaughterhouse ovaries by slicing the surface of the ovary. Only oocytes with a compact cumulus investment were used. Oocytes were placed into three groups: (1) control--placed immediately into culture; (2) holding control--COCs kept in PBS containing 0.4% BSA for 90 min before placement into culture; and (3) treatment--incubation with BCB for 90 min before culture. Treated oocytes were then divided into BCB- (colorless cytoplasm, increased G6PDH) and BCB+ (colored cytoplasm, low G6PDH) on their ability to metabolize the stain. Activity of G6PDH was determined via measurement of NADP reduction induced by G6P as substrate oxidized by G6PDH in the cytosol of control, BCB- and BCB+ groups; G6PDH activity was significant higher in BCB- COCs than in control and BCB+ COCs. After IVM, oocytes were fertilized in vitro. Embryos were cultured to day 8. The rate of maturation to metaphase II was significantly higher for control and BCB+ oocytes than for BCB- oocytes. The BCB+ oocytes yielded a significantly higher proportion of blastocysts (34.1%) than did control or holding control oocytes (18.3 and 19.2%); and both controls and BCB+ oocytes had significantly higher blastocyst development than did BCB- oocytes (3.9%). These results show that the staining of bovine cumulus oocyte complexes with BCB before in vitro maturation may be used to select developmentally competent oocytes for IVF. In addition, G6PDH activity may be useful as a marker for oocyte quality in future studies on factors affecting developmental competence. PMID- 15826684 TI - Effect of undegradable protein concentration in the post-weaning diet on body growth and reproductive development of Assaf rams. AB - Twenty-four growing Assaf lambs, divided into four groups of six animals, were used to study the effect of the undegradable protein content of the post-weaning diet on feed intake, body growth and reproductive development. From week 1 to week 21, the four groups were fed ad libitum as follows: group LL was given barley straw and low protein concentrate (LP); group HH was given barley straw and high protein concentrate (HP); group LH was given barley straw and LP concentrate from week 1 to 11 (period 1) and barley straw and HP concentrate from week 12 to 21 (period 2); group HL was given barley straw and HP concentrate in period 1 and barley straw and LP concentrate in period 2. From week 22 to week 26 (period 3), all animals received the same amount of hay and LP concentrate. Barley straw intake was not significantly (P>0.05) affected by dietary treatments. In the 1st period, average concentrate intake and live body weight gain (LWG) were greater in lambs fed HP than LP supplement. In the 2nd period, concentrate intake was not significantly (P>0.05) affected by type of supplement, but LWG was greater for lambs fed HP than LP supplement. Scrotal circumference in week 11 was significantly (P<0.05) lower in lambs fed LP supplement than in lambs fed HP supplement. No significant differences (P>0.05) due to dietary treatments were observed on scrotal circumference in weeks 21 and 25. Dietary treatments had no significant (P>0.05) effect on either circulating concentration of testosterone or ejaculate characteristics. In conclusion, results from this study suggest that supplementing diets with undegradable protein enhanced performance throughout the breeding period and accelerated testis growth. Nevertheless, final testis size, pattern of circulating testosterone and sperm output were unaffected by dietary treatments. PMID- 15826685 TI - Effects of a high semen-collection frequency on the quality of sperm from ejaculates and from six epididymal regions in boars. AB - This study examines the effect of semen-collection rhythm on the sperm maturation process in boar epididymis. Three post-pubertal boars were submitted to a high semen-collection frequency (stressed boars) during 4 days, and three males were kept as a control group (control boars). Semen samples coming from six epididymal regions and from the ejaculate of each male were evaluated for sperm concentration, sperm vitality, sperm motility and sperm morphology. In each epididymal region, either fluid resorption or fluid secretion was determined from the variation in sperm concentration. The pattern of fluid resorption-secretion along the epididymal duct differed significantly between the stressed and control boars. A high semen-collection frequency also affected the development of sperm motility and the sperm cytoplasmic droplet displacement along the epididymal duct. The incidence of some sperm abnormalities was also found to be higher in some epididymal regions and ejaculates of stressed boars. From the results of this study, it can be concluded that a high semen-collection frequency brings about an altered resorption and secretion pattern of the epididymal fluid, which results in defective sperm maturation and abnormal development of sperm motility. PMID- 15826686 TI - Culture of early stage ovine embryos to blastocyst enhances survival rate after cryopreservation. AB - The current study assessed both the effects of in vitro culture and developmental stage of early stage in vivo produced ovine embryos on their ability to survive cryopreservation. Early stage embryos (n=226) were recovered from the oviduct, at different days of the early luteal phase, at three different developmental stages: 2- to 4-cell, 5- to 8-cell and 9- to 12-cell. For each stage, half of the embryos were cultured to the blastocyst stage and frozen thereafter (CF), while the remainder was frozen just after recovery (EF). A third experimental group (BF; n=43) included blastocysts obtained from the uterus and frozen immediately after recovery. Embryo viability post-thawing was determined by assessing their rate of development to the hatched blastocyst stage following in vitro culture. Culture negatively affected embryo viability, since survival rate was higher in blastocysts obtained from the uterus than in those from culture (83.7% versus 66.1%; P<0.05); also the cryosurvival of cultured embryos was lower when the timing of blastocyst formation was extended (P<0.01). However, survival following freezing-thawing of early developmental stages was significantly lower when compared to viability of their counterparts cultured to the blastocyst stage (23.1% versus 66.1%, P<0.0001). In conclusion, our results indicate that, despite the deleterious effects of culture per se, the culture of early in vivo produced ovine embryos to the blastocyst stage prior to be frozen improves their survival after thawing. PMID- 15826687 TI - Benefits of TEMPOL on ram semen motility and in vitro fertility: a preliminary study. AB - Extending the preservation time of fresh semen is an important goal in artificial insemination programs particularly for ewes in natural oestrus, where insemination periods are longer than for ewes synchronized with hormonal treatments. The aim of this study was to evaluate the effect of the antioxidant TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) on the maintenance in long term storage of ram semen motility and fertility. Semen from Sarda breed rams was diluted in two extenders: sodium citrate buffer with TEMPOL and skimmed milk, used as control. Samples diluted with TEMPOL were cooled at either 15 degrees C or 22 degrees C, while those diluted with skimmed milk were cooled at 15 degrees C. Each sample was divided into four stocks, and stored for different times (5 min, 24, 48 and 72 h). Three aliquots were taken from each stock for every storage period. One was immediately evaluated under microscope; one was used for in vitro fertilization; one was incubated for 2 h in controlled humidified atmosphere (5% CO2, 7% O2 and 88% N2) at 39 degrees C, then evaluated for motility and utilized for in vitro fertilization. Ram semen diluted with media containing TEMPOL demonstrated increased motility, fertility and an improved protective effect when it was stored at 15 degrees C. PMID- 15826688 TI - Differences in the incidence of apoptosis between in vivo and in vitro produced blastocysts of farm animal species: a comparative study. AB - The occurrence of pregnancies and births after embryo transfer (ET) of in vivo produced embryos is generally more successful compared to that of embryos produced in vitro. This difference in ET success has been observed when embryos of morphological equal (high) quality were used. The incidence of apoptosis has been suggested as an additional criterion to morphological embryo evaluation in order to assess embryo quality and effectively predict embryo viability. In this study, equine, porcine, ovine, caprine and bovine in vivo and in vitro produced morphologically selected high quality (grade-I) blastocysts were compared for the occurrence of apoptosis in blastomeres. The total number of cells per embryo and the number of cells with damaged plasma membranes, fragmented DNA and fragmented nuclei per embryo were assessed in selected blastocysts by combining Ethidium homodimer (EthD-1), terminal dUTP nick end labeling (TUNEL) and Hoechst 33342 staining. In general, the level of blastomere apoptosis was low. A higher level of apoptosis was observed in in vitro produced equine, porcine and bovine blastocysts compared to their in vivo counterparts. Interestingly, 4 of the initially selected 29 bovine in vitro produced blastocysts exhibited extensive signs of apoptosis affecting the inner cell mass (ICM), which is not compatible with a viable conceptus. Repeated occurrence of this observation may explain the lower ET outcome of in vitro produced bovine embryos compared to in vivo produced embryos. It is concluded that, although in morphologically high quality blastocysts of several farm animal species a significant difference exists in the percentages of apoptotic cells between in vivo and in vitro produced embryos, the incidence of apoptosis at the blastocyst stage is at such a low level that it cannot reflect the substantial differences in embryo viability that have been described between in vivo and in vitro produced blastocysts following ET. PMID- 15826689 TI - Production of piglets with sexed semen employing a non-surgical insemination technique. AB - The aim of the present study was to ascertain whether multiparous sows could successfully be inseminated with sexed semen non-surgically. Spermatozoa were stained with Hoechst 33342 and separated flowcytometrically in X- and Y chromosome bearing sperm populations employing the Beltsville Sperm Sexing Technology (BSST). After weaning, estrus was induced in sows with PMSG and hCG. Animals were inseminated once per estrus non-surgically with a specially designed catheter into the tip of the uterine horn, employing 50x10(6) of either sexed or non-sexed spermatozoa diluted in 2 ml Androhep. Pregnant sows were allowed to go to term. Mean pregnancy rate from inseminations with unsexed spermatozoa was 54.5% whereas inseminations with sexed spermatozoa resulted in 33.3% pregnant sows. All but one piglet born after insemination with sexed semen were of the predicted sex. The sex of those piglets born after inseminations with non-sexed spermatozoa was 61.1% for male and 38.9% for female sex. It is concluded that non surgically inseminations with flowcytometrically sexed spermatozoa can be conducted successfully. PMID- 15826690 TI - Detection of cooling-induced membrane changes in the response of boar sperm to capacitating conditions. AB - There is a need for methods of rapid and sensitive sperm function assessment. As spermatozoa are not able to fertilize an oocyte before having undergone a series of complex physiological changes collectively called capacitation, it is logical to assess sperm function under fertilizing conditions in vitro. In this study, the responsiveness of sperm to capacitating conditions in vitro was monitored by changes in sperm response to ionophore and by changes in the amount of intracellular calcium ions in stored boar semen. Boar semen was diluted at 32 and 20 degrees C and stored for 24 and 72 h at 16 and 10 degrees C. Ionophore-induced changes and increased intracellular calcium ion content in boar spermatozoa were recorded by flow cytometry and found to progress as a function of time during incubation under capacitating conditions. All responsiveness parameters (increases in proportions of membrane-defective spermatozoa, acrosome-reacted spermatozoa, and cells with high intracellular calcium levels) were shown to be sensitive to subtle physiological changes occurring at low storage temperatures. The initial levels of sperm with a high calcium content were higher in semen stored at 10 degrees C, but the accumulation of internal calcium was lower than in semen stored at 16 degrees C. The loss of membrane integrity and increase in the proportion of acrosome-reacted cells were higher in semen stored at 10 degrees C. Dilution at 20 degrees C had no negative effect on membrane integrity or responsiveness to capacitating conditions. There was no significant difference between semen stored for 24 and 72 h in terms of membrane integrity, acrosome reaction, and intracellular calcium after capacitation treatment. However, dynamics of cell death and acrosome reaction in response to capacitating conditions were somewhat accelerated after 72 h storage, especially in semen stored at 10 degrees C. It can be concluded that the simultaneous use of the sperm membrane responsiveness and kinetic parameters is a sensitive tool for the detection of storage-related membrane changes in boar semen. PMID- 15826691 TI - Influence of cysteamine supplementation and culture in portable dry-incubator on the in vitro maturation, fertilization and subsequent development of mouse oocytes. AB - The need to transport oocytes and embryos between two laboratories have prompted us to evaluate the effects of in vitro maturation of immature mouse oocytes in a CO2-deficient dry heat portable incubator and subsequent in vitro development of these fertilized mouse oocytes in a standard CO2 incubator. In addition, the effects of cysteamine supplementation on maturation rate and embryonic development during in vitro maturation (IVM) and culture of embryos in the portable incubator were also investigated. Germinal vesicle stage mouse oocytes, recovered at 40-h post-FSH from 6- to 8-week-old C57BL/6xCBA F1 healthy female mice, were matured in vitro in a modified TCM-199 supplemented with or without 100 microM cysteamine in a standard incubator (5% CO2; 37 degrees C) or cultured in a CO2-deficient dry heat portable incubator for 5 h at 37 degrees C and thereafter transferred to a standard incubator for further culture. The addition of cysteamine in the IVM medium significantly improved maturation rates of the GV mouse oocytes to metaphase II stage. However, cysteamine supplementation in the culture medium did not significantly improve fertilization and blastocyst formation rates of IVM and ovulated oocytes, and in vivo-derived zygotes. Culture conditions in a CO2-deficient dry heat portable incubator did not adversely affect the developmental competence of in vivo-derived zygotes and in vitro matured mouse oocytes after IVF or parthenogenetic activation. Cysteamine supplement in the IVM medium could enhance nuclear maturation of these immature oocytes during shipment. PMID- 15826692 TI - Mitochondrial activity of frozen-thawed spermatozoa assessed by MitoTracker Deep Red 633. AB - The present study was conducted to find a more objective method of evaluating sperm quality than the current subjective motility evaluations by testing the applicability of a novel fluorescent probe, Mitotracker Deep Red 633 (M-22426), for measuring the mitochondrial activity of spermatozoa both after freezing/thawing (PT) and after swim-up selection (SU), using flow cytometry (FC). The results from FC were compared to those of conventional microscopic motility evaluations and of computer-assisted sperm analysis (CASA) as well as to the fertility obtained after AI in the field. Semen from six Estonian Holstein bulls, processed when the sires were aged 3, 5, and 7 years, was included in the experiment. Sperm motility (measured either subjectively or by means of CASA) was always significantly (p<0.01-0.001) higher in the spermatozoa recovered by SU, for any of the age groups considered, or even when combining the age groups. Motility (measured subjectively) after SU was significantly (p<0.05) higher when bulls reached 7 years of age, compared to earlier collection ages, but no differences were registered between ages for CASA-assessed motility, either after SU or immediately PT. The numbers of spermatozoa with high red fluorescence also increased after SU: p<0.05 (for semen from bulls aged 3 years), p<0.001 (5 years), p<0.001 (7 years), and p<0.001 when all age groups were combined. The proportion of spermatozoa with high mitochondrial activity as determined by Mitotracker Deep Red 633 correlated with sperm motility (p<0.01) both PT and after SU, but not with the fertility results. In conclusion, MitoTracker Deep Red 633 seems to be a reliable marker for frozen-thawed bovine semen viability both PT and after SU. PMID- 15826693 TI - Treating heifers with GnRH from 4 to 8 weeks of age advanced growth and the age at puberty. AB - In heifer calves, an early transient increase in circulating concentrations of LH is associated with early follicular development and is thought to regulate the timing of puberty. In an attempt to hasten the onset of sexual maturity, the early rise in LH concentration was advanced by injecting heifer calves with 120 ng/kg of GnRH (n=6) twice daily from 4 to 8 weeks of age; control calves received saline (n=6). Blood samples were collected every 15 min for 10h at 4, 8, 14, 20, 26, 32, 38, 44 and 50 weeks of age. Treatment with GnRH increased mean circulating concentrations of LH at 8 weeks of age (P<0.05), LH pulse frequency at 4 and 8 weeks of age (P<0.05), and reduced the mean age at puberty by 6 weeks (56.8+/-1.7 versus 62.8+/-2.4 weeks of age, for GnRH treated and control calves, respectively; P=0.04). Body weight gain was greater in GnRH-treated calves than control calves (P<0.05), and the rate of weight gain was shown to be a significant covariate within age at puberty. In conclusion, we suggest that the timing of the early rise in LH concentrations is a critical signal involved in the timing of puberty in heifers. PMID- 15826694 TI - Embryonic mortality in buffaloes synchronized and mated by AI during the seasonal decline in reproductive function. AB - The aim was to determine the factors that contribute to embryonic mortality in buffaloes mated by AI during a period of increasing day length which corresponds to a natural decline in reproductive activity. Italian Mediterranean buffalo cows (n=243) showing regular estrous cycles were synchronized using the Ovsynch-TAI program and mated by AI at 16 and 40 h after the second injection of GnRH. Blood samples were collected on Days 10 and 20 after the first AI and assayed for progesterone (P4). Pregnancy diagnosis was undertaken on Days 26 and 40 after the first AI using rectal ultrasonography. Buffaloes with a conceptus on Day 26 but not on Day 40 were judged to have undergone embryonic mortality and for these animals uterine fluid was recovered by flushing and analysed for common infectious agents. Estrus synchronization was achieved in 86% of buffaloes and the pregnancy rate on Day 40 was 34%. Embryonic mortality between Days 26 and 40 occurred in 45% of buffaloes and was associated with the presence of significant infectious agents in only 10 buffaloes (8%). Concentrations of P4 on Day 10 after AI were higher (P<0.05) in buffaloes that established a pregnancy than in buffaloes that showed embryonic mortality that was not associated with infectious agents. Similarly, on Day 20 after AI P4 concentrations were higher (P<0.01) in pregnant buffaloes compared with non-pregnant buffaloes and buffaloes that had embryonic mortality. It is concluded that a reduced capacity for P4 secretion can explain around 50% of embryonic mortalities in buffaloes synchronised and mated by AI during a period of low reproductive activity and that other as yet unidentified factors also have a significant effect on embryonic survival. PMID- 15826695 TI - Influence of force location in orthodontic shear bond strength testing. AB - OBJECTIVE: The purpose of the present study was to analyze the influence of debonding force location in shear bond strength testing of orthodontic brackets in vitro. METHODS: Ninety extracted permanent bovine mandibular incisors were randomly divided into 3 groups of 30 specimens each. Teeth were bonded with stainless steel orthodontic brackets. Enamel surfaces were etched with 37% phosphoric acid for 30s and bonded with a composite adhesive. Debonding force measurements were performed with a universal testing machine. Location of the debonding force was: bracket base (group A), ligature groove (group B), occlusal bracket wings (group C). RESULTS: Mean shear bond strength measurements were as follows: 22.70(4.23)MPa (group A), 11.52(2.74)MPa (group B), 9.44(2.96)MPa (group C). Analysis of variance indicated that there were significant differences in shear bond strength. Post-hoc Tukey tests showed that bond strength measurements of group A were significantly different from those of groups B and C. The adhesive remnant index also showed significant differences and ranged from a mean of 1.53 in group A to a mean of 2.10 in group C. SIGNIFICANCE: Debonding force location had a significant influence on shear bond strength measurements and bond failure pattern, indicating that this parameter needs to be taken into consideration for interstudy comparison of in vitro results. PMID- 15826696 TI - Self-etch vs etch-and-rinse adhesives: effect of thermo-mechanical fatigue loading on marginal quality of bonded resin composite restorations. AB - OBJECTIVE: This study evaluated the marginal integrity of dentine adhesives bonded to enamel and dentine, before and after thermo-mechanical loading (TML). METHODS: MO cavities with proximal boxes beneath the CEJ were prepared in extracted human third molars. Direct resin composite restorations (Tetric Ceram) were bonded with 3-step etch-and-rinse (Syntac Classic, Solobond Plus, OptiBond FL), 2-step etch-and-rinse (Admira Bond, Single Bond), 2-step self-etch (AdheSE, Clearfil SE Bond), and 1-step self-etch (all-in-one) adhesives (Adper Prompt, Xeno III, iBond). Marginal gaps were analyzed using SEM of epoxy resin replicas. Bonded interfaces before TML were examined with TEM to identify pre-existing attributes for subsequent marginal disintegration. RESULTS: In enamel, high percentages of gap-free margins were initially identified for all adhesives. After TML, etch-and-rinse adhesives exhibited significantly higher percentages of gap-free margins (approximately 90%) compared with two-step self-etch (approximately 75%) and all-in-one (approximately 55%) adhesives (p<0.05). iBond did not completely etch through the enamel smear layer. In dentine, 89-100% gap free margins were initially observed. After TML, there were no statistical differences among etch-and-rinse (62-70%) and two-step self-etch (62-63%) adhesives (p>0.05). The all-in-one adhesives exhibited significantly less gap free margins (<40%) in dentine (p<0.05), with iBond showing the worst marginal integrity (15%). The presence of pre-existing water channels within the adhesives probably expedited water sorption when restorations were under functional stresses. CONCLUSION: Enamel bonding was more effective with phosphoric acid etching. Etch-and-rinse and 2-step self-etch adhesives showed promising marginal adaptation to dentine and may have a better clinical prognosis than the all-in one bonding approach. PMID- 15826697 TI - Effect of fracture strength of primer-adhesive mixture on bonding effectiveness. AB - OBJECTIVE: The purpose of this study was to measure the micro-tensile bond strength (microTBS) of two adhesives to dentin and to correlate this microTBS with the micro-tensile fracture strength (microTFS) of primer-adhesive mixtures of the two adhesives. METHODS: The three-step etch&rinse adhesive OptiBond FL (Kerr) and the two-step self-etch adhesive Clearfil SE Bond (Kuraray) were used to bond Filtek Z100 (3M ESPE) to twenty mid-coronal dentin surfaces. The microTBS was determined at 1 and 24 h after light-curing. In addition, the microTFS of primer-adhesive mixtures (primer:adhesive=1:3 by weight) and of the pure adhesives (controls) were measured after the same time intervals. RESULTS: The microTBS to dentin of OptiBond FL at 24 h was significantly higher than at 1 h and than the microTBS of Clearfil SE Bond at 24 h. The mean microTFS of the primer-adhesive mixtures was always lower than the microTFS of the respective adhesives. The highest reduction in microTFS due to mixing primer with the adhesive resin was measured for Clearfil SE Bond. Both the microTFS of the primer adhesive mixtures and the pure adhesives of OptiBond FL and Clearfil SE Bond increased with time. SIGNIFICANCE: The early 1 h microTBS to dentin was lower than the 24 h microTBS for OptiBond FL, but not for Clearfil SE Bond. Differences in polymerization efficiency as well as in the resulting interfacial strength may explain such varying early bond strengths to dentin. PMID- 15826698 TI - The static strength and modulus of fiber reinforced denture base polymer. AB - OBJECTIVES: Partial fiber reinforcements have been employed to strengthen dentures both during repair and in the manufacturing process. The reinforcing fibers can be evenly distributed in the denture base polymer or alternatively fiber-rich phase in the denture base polymer can form a separate structure. The aim of this study was to determinate static three-point flexural strength and modulus of denture base polymer that had been reinforced with different fiber reinforcements. METHODS: The test specimens (3 x 5 x 50 mm) were made of auto polymerized denture base polymer and reinforced with different fiber reinforcements. The test groups were: (A) no fibers; (B) non-impregnated polyethylene fibers; (C) light-polymerized monomer impregnated glass fibers; (D) porous polymer preimpregnated glass fibers and (E) light-polymerized monomer polymer impregnated glass fibers. The fibers were oriented parallel to the long axis of the specimen and embedded into the denture base resin on the compression side (n=7) or tension side (n=7). Dry specimens were tested with three-point static flexural strength test set-up at crosshead speed of 5 mm/min. RESULTS: The statistical analysis by two-way analysis of variance showed that the brand and the location of the fiber reinforcements significantly influenced the flexural strength (p<0.0001). However, the location of the fiber reinforcements did not influence the flexural modulus (p<0.722). SIGNIFICANCE: The results suggest that impregnated and preimpregnated fibers reinforce denture base polymer more than non-impregnated fibers. Fiber reinforcements placed on the tensile side resulted in considerably higher flexural strength and flexural modulus values compared with same quantity of fibers placed on the compression side. PMID- 15826699 TI - Anisotropic fracture in bovine root and coronal dentin. AB - OBJECTIVES: This purpose of this study was to address the fundamental question of how the fracture properties of dentin are related to its composite structure. METHODS: Strain concentration tests and impression-induced damage tests were designed to compare bovine root dentin with coronal dentin, and to understand the role of individual structural elements in the fracture of dentin. RESULTS: Absent in peritubular dentin, root dentin is insensitive to cracks and exhibits higher fracture resistance than coronal dentin that has a typical brittle fracture behavior along the peritubular dentin. Fracture analysis and impression damage experiments found that root dentin is highly anisotropic in fracture behavior. Cracking is predominantly controlled by the organization of collagen fibrils, with the incremental lines being the weakest planes. In coronal dentin, highly mineralized peritubular dentin that intersects with the incremental lines creates additional weak orientations that compete with the incremental lines and thus greatly decrease the degree of fracture anisotropy. SIGNIFICANCE: This study demonstrated that dentin is by no means homogeneous in terms of fracture properties. Location and orientation (especially in terms of incremental lines) should be taken into account when examining tooth failure both in laboratory and in clinical studies. PMID- 15826700 TI - The adhesion between prefabricated FRC posts and composite resin cores: microtensile bond strength with and without post-silanization. AB - OBJECTIVES: Aim of the study was to measure the adhesion between two types of translucent prefabricated FRC posts (FRC Postec, Ivoclar-Vivadent, FRC; Light Post, RTD, LP), and two types of flowable composites used as core materials (UnifilFlow, GC, UF; Tetric Flow, Ivoclar-Vivadent, TF), with or without the application of a silane (Monobond-S, Ivoclar-Vivadent, S) on the post surface. METHODS: The experimental groups were: 1.1 FRC+UF; 1.2 FRC+S+UF; 1.3 FRC+TF; 1.4 FRC+S+TF: 2.1 LP+UF; 2.2 LP+S+UF; 2.3 LP+TF; 1.4 LP+S+TF. The bond strength at the interface between post and core was measured with the microtensile non trimming technique. Thirty to thirty-five beam-shaped specimens per group were obtained from cylinders of core material, which had been built up around the post by progressively adding small increments of composite resin. Each specimen was loaded in tension until failure at either one of the two post-core interfaces present in each stick. The differences in interfacial bond strength among the groups were tested for statistical significance with the two-way ANOVA. RESULTS: The measured bond strengths in MPa were: [table: see text]. The statistical analysis revealed that post-silanization had a significant effect on adhesion (p<0.05). With any combination of post and core materials tested, the application of a silane onto the post surface prior to building up the core significantly increased the post-core bond strength. SIGNIFICANCE: For improved adhesion at the interface between prefabricated FRC posts and composite resin cores, post silanization is advisable. PMID- 15826701 TI - Microleakage of various cementing agents for full cast crowns. AB - OBJECTIVES: To evaluate microleakage and marginal gaps in full cast crown restorations bonded with six different types of cementing agents. METHODS: Sixty non-carious human premolars and molars were prepared in a standardized manner for full cast crown restorations. The mesial and distal margins were located in dentin, while the vestibular and palatal/lingual margins were located in enamel. Crowns were made from a high-gold alloy using a standardized technique. The specimens were randomized to six groups of cementing agents: one zinc-phosphate cement (Harvard cement), one conventional glass-ionomer cement (Fuji I), one resin-modified glass-ionomer cement (Fuji Plus), two standard resin cements (RelyX ARC, Panavia F), and one self-adhesive universal resin cement (RelyX Unicem). After 4 weeks of storage in distilled water at 37 degrees C, the specimens were subjected to 5000 thermocycles ranging from 5 to 55 degrees C. Then, they were placed in a silver nitrate solution, embedded in resin blocks, and vertically cut in buccolingual and mesiodistal direction. Subsequently, the objects were evaluated for microleakage and marginal gap using a high-resolution digital microscope camera. RESULTS: A number of inter-group differences were statistically significant. RelyX Unicem showed the smallest degree of microleakage both in enamel and in dentin. Panavia F und RelyX Unicem were associated with significantly larger marginal gaps than all other cementing agents. No association was observed between microleakage and marginal gap other than a weak direct correlation when using Harvard cement on enamel. SIGNIFICANCE: The cementing agents investigated revealed different sealing abilities. These differences were not associated with specific types of materials. PMID- 15826702 TI - Influence of surface and heat treatments on the flexural strength of a glass infiltrated alumina/zirconia-reinforced dental ceramic. AB - OBJECTIVES: The purpose of this study was to investigate the influence of sandblasting, grinding, grinding orientation and polishing before and after heat treatment, on the flexural strength of a glass-infiltrated alumina/zirconia reinforced dental ceramic (In-Ceram Zirconia). METHODS: The uniaxial flexural strength was calculated on 160 bar-shaped specimens (20 x 4 x 1.2mm) divided equally into eight groups as follows: sandblasted; sandblasted heated; polished; polished heated; ground parallel to the tensile axis; ground parallel heated; ground perpendicular; and ground perpendicular heated. Data were analyzed with multiple regression analysis, one-way ANOVA and Tukey's pairwise multiple comparisons and Weibull analysis. The treated and fractured surfaces were observed with SEM. The relative content of the monoclinic phase was quantified with an X-ray diffraction analysis. RESULTS: A thin layer of glass was present on the surface of the specimens after heat treatment and contributed to an improvement of the flexural strength. Surface treatment (not followed by heat treatment) generated phase transformation which, however, was not sufficient to avoid strength degradation caused by the flaws introduced with the surface treatments. Sandblasting caused the most marked strength degradation. Polishing alone increased the reliability, but did not improve the strength. The orientation of grinding in respect of the direction of the tensile stresses did not influence the ultimate tensile strength. SIGNIFICANCE: The present study suggests that any surface treatment performed on In-Ceram Zirconia should always be followed by heat treatment to avoid strength degradation. PMID- 15826703 TI - Effect of two different bleaching regimens on the gloss of tooth colored restorative materials. AB - OBJECTIVES: Vital tooth bleaching with peroxide is one of the most common cosmetic procedures in dentistry and can be accomplished using a variety of methods or regimens. Recently, new generation of tooth color restorative materials were introduced to market. The purpose of this in vitro study was to determine the gloss changes of three different tooth color restorative materials: Flowable composite (Filtek Flow/3M), packable composite (Filtek P60/3M) and ormocer (Definite/DEGUSSA) after two different bleaching regimens (Vivastyle/VIVADENT) and (Crest Professional Whitestrips/PROCTER and GAMBLE). METHODS: 16 specimens 30 x 30 x 2 mm size were fabricated from each restorative material. After gloss values were measured with gloss meter, at two different angles of illumination (20 and 60 degrees ), 10% carbamide peroxide (Vivastyle) was applied for 2 h per day for fourteen days to the half of the specimens while 6.5% hydrogen peroxide strip bands (Crest Professional Whitestrips) were applied to the remaining eight of the specimens for 30 min twice daily for 14 days. During the test period the specimens were stored in 37 degrees C and 100% relative humidity. At the end of bleaching regimen the gloss measurements were repeated and the data were subjected to statistical analysis. RESULTS: The Wilcoxon Signed Rank Test analysis revealed that the gloss values were affected by both bleaching regimens (P=0.012). Whitestrips decreased the gloss values of Filtek P60 (at 20 and 60 degrees , P<0.001) and Filtek Flow (at 20 degrees , P=0.05 and at 60 degrees , P=0.05) significantly compared to Vivastyle, while the gloss values of Definite did not show any significant change between Vivastyle and Whitestrips application (at 20 degrees P=0.279; at 60 degrees , P=0.279, Mann Whitney U Test). The gloss values of materials were significantly different before (at 20 degrees P<0.001; at 60 degrees , P=0.003) and after bleaching (at 20 degrees P<0.05; at 60 degrees , P<0.05) with the highest value of Filtek Flow followed Filtek P60 and then by Definite (Kruskal Wallis test). SIGNIFICANCE: As the gloss of tooth colored restorative materials could be affected by bleaching regimens, it is necessary to consider the type of the material before starting the treatment. PMID- 15826704 TI - The degree of conversion of fiber-reinforced composites polymerized using different light-curing sources. AB - OBJECTIVES: Fiber-reinforced composites (FRC) with different polymer matrix compositions are available on the market. The aim of this study was to compare the degree of monomer conversion of FRC that contained either dimethacrylate resin matrix or semi-IPN resin matrix. In addition, the effect of different types of light-curing units and durations of light irradiation on the degree of conversion was evaluated. MATERIALS AND METHODS: Continuous unidirectional fibers impregnated with dimethacrylate or semi-IPN resin were compared. As control materials, a particulate filler composite resin and an unfilled resin were used. The materials were photopolymerized using various irradiation times and different types of light-curing units (quartz-tungsten-halogen with standard or turbo tip, light-emitting diode and plasma-arc). Degree of monomer conversion (DC%) was determined by FT-IR spectrometry. The infrared spectra were recorded at seven different time points after polymerization. RESULTS: ANOVA showed significant differences between the materials tested, and the curing units (P<0.05). The DC% of semi-IPN and dimethacrylate matrix FRC was not statistically different (P>0.05) compared to that of unfilled resin. Conventional quartz-tungsten halogen with turbo tip showed the highest DC% (69-72.3%), whereas the plasma-arc curing unit showed the lowest degree of conversion (47-62%). SIGNIFICANCE: Within the limitations of this study, it can be concluded that semi-IPN matrix of FRC did not show a difference in the degree of conversion compared to dimethacrylate resin FRC when quartz-tungsten or a light-emitting diode polymerization unit was used. The plasma-arc light-curing unit showed a lower degree of conversion with all materials compared to the light-emitting diode and quartz-tungsten-halogen lamps. PMID- 15826705 TI - Fracture resistance of yttrium oxide partially-stabilized zirconia all-ceramic bridges after veneering and mechanical fatigue testing. AB - OBJECTIVES: Yttria-stabilized zirconia (Y-TZP) ceramic is a high-performance material with excellent mechanical properties suitable for fixed partial dentures. The purpose of this study was to evaluate after fatigue testing, the effect of heat-treatment and veneering on the fracture resistance of frameworks manufactured using sintered and subsequently hot isostatic pressed yttrium oxide partially-stabilized zirconia (Denzir). METHODS: The specimens were subjected to dynamic loading in water. Thereafter, using a universal testing machine, the fracture resistance of three-unit fixed partial dentures was determined; (i) of the frameworks as delivered after machining, (ii) after the frameworks had been subjected to heat-treatment similar to veneering with a glass-ceramic (Eris) or a feldspar-based ceramic (Vita D) and (iii) after the frameworks had been veneered with these ceramics. In addition, the fracture resistance of frameworks as delivered after machining not subjected to dynamic loading in water was determined. RESULTS: Cyclic loading in water did not significantly (p>0.05) affect the fracture resistance. The load necessary to fracture the frameworks as delivered after machining was significantly (p<0.001) higher than for the heat treated and veneered specimens. No significant difference was seen between the specimens veneered with Eris and those veneered with Vita D (p>0.05). For all but three specimens the fractures were located in the loading point and through one of the connectors. SIGNIFICANCE: Heat-treatment and veneering reduce fracture resistance of hot isostatic pressed zirconia. Nevertheless, the results obtained indicate that it is an interesting material for potential in all-ceramic restorations. PMID- 15826706 TI - Shrinkage behavior of a resin-based composite irradiated with modern curing units. AB - OBJECTIVE: The present study determined the influence of different light curing regimes (four light-emitting diode (LED) units (Freelight 1 and 2, 3M-ESPE; e light, GC; Bluephase (prototype), Ivoclar Vivadent), two quartz-tungsten-halogen (QTH) lights (Astralis 10, Ivoclar Vivadent; Swiss Master Light, EMS) and one plasma-light curing unit (Easy Cure, DMDS)) on the curing behavior of a resin based composite material (InTen-S, Ivoclar Vivadent). METHODS: Polymerization shrinkage was induced by light curing the tested material with 14 different regimes of the curing units mentioned above. The contraction stress was recorded for 300 s at room temperature with a Stress-Strain-Analyzer (c(FACTOR)=0.3). The maximum contraction stresses after 300 s, the time until gelation (t(0.5N)), and the coefficient of near linear fit of contraction force/time (gradient) were analyzed. The statistical analysis was conducted using ANOVA (alpha=0.05) and Tukey's post hoc test. RESULTS: The five tested regimes of the LED unit e-light revealed the lowest statistically significantly maximum contraction stress followed by the low intensity LED unit Freelight 1 and the plasma curing unit Easy Cure. The high intensity LED unit Freelight 2 exhibited a significantly higher contraction stress compared to Freelight 1. No significant differences between the standard and exponential modes within these curing units were found. No significant differences were found between the LED unit Freelight 2 and the pulse program of the halogen light curing unit Astralis 10. The highest polymerization stresses were observed for the high energy curing units, either QTH (Swiss Master Light and Astralis 10) or LED (Bluephase). SIGNIFICANCE: Fast contraction force development, high contraction stress and an early start of stress build-up cause tension in the material with possible subsequent distortion of the bond to the tooth structure. The lowest polymerization stress was observed for the low energy LED lamps, while the plasma unit and the high energy QTH and LED curing units produced two to three times higher stress. PMID- 15826708 TI - Treatment for hepatic encephalopathy: tips from TIPS? PMID- 15826709 TI - Refined tools for the treatment of hepatocellular carcinoma. PMID- 15826710 TI - Treatment with peg-interferon alfa-2b and ribavirin of hepatitis C virus associated mixed cryoglobulinemia: a pilot study. AB - BACKGROUND/AIMS: The aim of this study is to verify the efficacy and safety of peg-interferon alfa-2b in combination with ribavirin for initial treatment of HCV associated mixed cryoglobulinemia. METHODS: Eighteen patients (7 women and 11 men) affected by mixed cryoglobulinemia were included in the study and treated with peg-interferon alfa-2b 1.0 microg/kg once a week plus ribavirin (1000 mg daily) for 48 weeks, regardless of the HCV genotype. RESULTS: At the end of the treatment HCV-RNA became undetectable in 15 patients (83%) and most patients improved clinically. One subject suspended treatment at 13th week due to depression. A large fraction of the patients (8 cases: 44%) relapsed both virologically and clinically a few weeks after the end of therapy. At the end of follow-up, only eight patients (44%) obtained a sustained virological response. CONCLUSIONS: Peg-interferon alfa-2b in combination with ribavirin seems safe and useful for patients affected by mixed cryoglobulinemia, but not as effective as in patients with HCV-positive chronic hepatitis without cryoglobulinemia. PMID- 15826711 TI - Estimating the impact of hepatitis C virus therapy on future liver-related morbidity, mortality and costs related to chronic hepatitis C. AB - BACKGROUND/AIMS: Chronic Hepatitis C virus (HCV) infection is common and often produces a progressive disease. Some studies suggest that HCV related complications will increase in the future. Our aim was to estimate the future morbidity, mortality and costs of chronic HCV infection in a cohort of patients infected by HCV and to evaluate the impact of HCV therapy. METHODS: A mathematical model was used to project over the next 30 years, the HCV related complications and costs in a cohort of 419,895 infected patients representing the HCV infected population in Spain. The impact of HCV therapy with peginterferon and ribavirin in this population was also projected. RESULTS: A gradual decline in the infected population is expected in the future, however, the proportion of patients with cirrhosis will increase by up to 14% and morbidity associated with HCV infection by up to 10% by the year 2030 with a subsequent increment in HCV related costs. However, treating from 10 to 50% of the HCV population will result in a reduction of 6 and 26% in morbidity and 4 and 20% in mortality, respectively. The cost per year of life gained ranges from 6078 for a 29-year-old patient to 8911 for a 59-year-old patient. CONCLUSIONS: In the future, HCV infection mortality, morbidity and associated costs will increase. Treatment of the chronic HCV infected population can eradicate the infection, increase patients' survival and reduce the need for liver transplantation, making this a cost-effective strategy. PMID- 15826712 TI - Diagnosis of hepatitis C virus related acute hepatitis by serial determination of IgM anti-HCV titres. AB - BACKGROUND/AIMS: We investigated whether serial determination of IgM to HCV core protein (HCV IgM) may be useful to identify acute hepatitis C (AHC) and to distinguish this disease from reactivation of chronic hepatitis C (r-CHC). METHODS: We enrolled 35 consecutive patients with AHC identified by seroconversion to anti-HCV and 31 consecutive patients who had been anti-HCV positive for at least six months at the time of reactivation. Titres of HCV IgM were calculated as Index values by a commercially available enzyme immunoassay. RESULTS: During the early phase of the illness we observed a wide variation in the HCV IgM Index values in all patients in the AHC group and consistent values in all cases in the r-CHC group. The HCV viral load determined soon after the onset of symptoms was of no use in identifying AHC.Twenty-three patients in the AHC group were observed as outpatients for 6-30 months; of these, 10 became plasma HCV-RNA negative within the third month of observation, but three showed a subsequent reactivation of HCV infection. CONCLUSIONS: Our data indicate that the detection of high and variable titres of HCV IgM in the early phase of the illness may identify acute hepatitis C and allow early antiviral treatment. PMID- 15826713 TI - Inhibition of glucagon improves splanchnic hyporesponse to terlipressin in cirrhotic rats with blood retention in the gastric lumen. AB - BACKGROUND/AIMS: Portal hypotensive effect of terlipressin is less effective when given during hemorrhage than in stable state. Blood retention in the stomach can induce splanchnic hyperemia which is mainly a consequence of an increased glucagon release. This study was undertaken to evaluate whether gastric blood retention contributes to the splanchnic hyporesponse to terlipressin. METHODS: Plasma glucagon determination was performed under basal conditions and after intragastric blood gavage in sham-operated and cirrhotic rats. Additionally, splanchnic hemodynamic effects to terlipressin were measured in blood-gavaged cirrhotic rats with or without glucagon antiserum or octreotide infusion. Another set of air-gavaged cirrhotic rats was included for comparison. RESULTS: Plasma glucagon level increased in both sham-operated and cirrhotic rats following blood gavage. Compared to air-gavaged cirrhotic rats, splanchnic hyporesponse to terlipressin was observed in cirrhotic rats receiving intragastric blood gavage. However, this splanchnic hyporesponse to terlipressin in blood-gavaged cirrhotic rats was overcome by glucagon antiserum or octreotide infusion. CONCLUSIONS: Intragastric blood gavage induced an elevation of plasma glucagon level and led to a splanchnic hyporesponse to terlipressin. Glucagon antiserum or octreotide administration overcame this hyporesponse. Excessive release of circulating glucagon may be an important factor for splanchnic hyporesponse to terlipressin in cirrhotic portal hypertension during hemorrhage. PMID- 15826714 TI - Clinical significance of extrapyramidal signs in patients with cirrhosis. AB - BACKGROUND/AIMS: Extrapyramidal signs have been described in cirrhosis and there is little information about their clinical significance. The aims of this study have been to investigate the relationship between extrapyramidal signs and cognitive impairment, and what is their influence on quality of life. METHODS: 46 patients with cirrhosis were evaluated for cognitive impairment with psychometric tests (Trail-Making Test part A, Grooved-Pegboard, Block-Design, Oral Symbol Digit and Stroop Test) and cognitive evoked potentials (P300). Extrapyramidal signs were evaluated using the UPDRS scale. Health-related quality of life (HRQL) was measured using SF-36 scale and the Chronic Liver Disease Questionnaire (CLDQ). RESULTS: Twenty-two patients had extrapyramidal signs, and these patients scored worse in all psychometric tests, except Block-Design. Patients with extrapyramidal signs also showed a longer P300 latency. Moreover, patients with extrapyramidal signs had the worst score in all the HRQL scales used. A multivariate analysis disclosed that the only variable showing an independent relationship to the mental component summary of SF-36 and with CLDQ was UPDRS score. CONCLUSIONS: We have found a clear relationship between the presence of extrapyramidal signs and cognitive impairment. Moreover, patients with extrapyramidal signs have worse scores in quality of life scales. PMID- 15826715 TI - High adiponectin in chronic liver disease and cholestasis suggests biliary route of adiponectin excretion in vivo. AB - BACKGROUND/AIMS: Models of fatty liver diseases and fibrosis suggest a hepatoprotective effect of adiponectin, an adipocyte-derived hormone with antidiabetic, antiobesity, antiatherogenic and anti-inflammatory effects. METHODS: We studied adiponectin serum levels in 111 chronic liver disease (CLD) patients and 226 healthy controls and the impact of cholestasis on adiponectin by bile duct ligation experiments in mice. RESULTS: Adiponectin was significantly elevated in CLD, and correlated with stage of liver cirrhosis, liver cell injury, e.g. aminotransferase activity, and inflammatory markers, but not with liver synthesis capacity, insulin sensitivity (HOMA index) or clinical complications. As patients with biliary liver diseases and cholestasis exhibited the highest adiponectin levels, we experimentally investigated a potential biliary route of adiponectin excretion. Following bile duct ligation in mice adiponectin levels rapidly increased without affecting hepatic adiponectin gene expression. Also, adiponectin was detectable in human bile. High adiponectin concentrations were associated with severe cholangitis and/or cholestasis on liver histology. CONCLUSIONS: Adiponectin is elevated in chronic liver disease and correlates with inflammation and liver damage. High adiponectin levels after bile duct ligation in mice and in human bile from cholestatic patients suggest that biliary secretion is involved in adiponectin clearance and that adiponectin could serve as a novel marker indicating cholestasis in liver cirrhosis. PMID- 15826716 TI - Pharmacological prophylaxis of hepatic encephalopathy after transjugular intrahepatic portosystemic shunt: a randomized controlled study. AB - BACKGROUND/AIMS: Hepatic encephalopathy is a frequent event after transjugular intrahepatic-portosystemic-shunt (TIPS), especially during the first months. Aim of this study was to compare two different treatments (lactitol 60 g/day, rifaximin 1200 mg/day) with no-treatment in the prevention of post-TIPS hepatic encephalopathy. METHODS: Seventy-five consecutive cirrhotics submitted to TIPS were randomized to receive either one of the above treatments or no-treatment. The main end-point was the occurrence of an episode of overt hepatic encephalopathy during the first month post-TIPS. Before the procedure and weekly thereafter the patients were evaluated by examining their mental status, asterixis, ammonia and trail-making-test Part-A (TMT-A). RESULTS: The three groups were comparable for age, sex, etiology, Child-Pugh-score, post-TIPS porto systemic gradient, previous hepatic encephalopathy, basal values of ammonia and psychometric performance. Twenty-five patients developed hepatic encephalopathy (33%, CI 95%=22-45%). One-month incidence was similar in the three groups (P=0.97). Previous hepatic encephalopathy (Relative Hazard=3.79;1.27-11.31) and basal-TMT-A Z-score>1.5 (RH=3.55;1.24-10.2) were predictors of post-TIPS encephalopathy at multivariate analysis. A <5 mmHg porto-systemic gradient was also significantly related to the occurrence of encephalopathy. CONCLUSIONS: Our data show that treatment with lactitol or rifaximin is not effective in the prophylaxis of hepatic encephalopathy during the first month after a TIPS. PMID- 15826717 TI - Blockade of intrahepatic adenosine receptors improves urine excretion in cirrhotic rats induced by thioacetamide. AB - BACKGROUND/AIMS: In healthy rats, we recently showed that reduced intrahepatic portal blood flow leads to activation of hepatic adenosine receptors and a nerve induced decrease in urine production. We hypothesize that the impaired urine excretion in liver cirrhosis is related to an increase in intrahepatic adenosine. METHODS: Anesthetized normal and thioacetamide-induced cirrhotic rats were instrumented for the measurement of urine flow, hepatic portal venous blood flow, and renal arterial blood flow. 8-Phenyltheophylline was used to block adenosine receptors. RESULTS: Compared to normal rats, cirrhotic rats had a lower baseline urine flow (P<0.05). In both normal and cirrhotic rats, intraportal but not intravenous administration of 8-phenyltheophylline increased urine flow. Saline overload in normal rats increased urine flow (from 6.8+/-0.6 to 42.2+/-4.6 microlmin(-1)) and this ability was impaired in cirrhotic rats (from 3.9+/-0.4 to 6.2+/-0.9 microlmin(-1)). Intraportal, but not intravenous, administration of 8 phenyltheophylline partially restored the renal ability to excrete the saline load. CONCLUSIONS: Impaired renal ability to excrete urine in liver cirrhosis is related to the activation of intrahepatic adenosine receptors, and this is consistent with our previous data showing renal regulation through a hepatorenal neural mechanism activated by intrahepatic adenosine. PMID- 15826718 TI - Combination of vitamin K2 and the angiotensin-converting enzyme inhibitor, perindopril, attenuates the liver enzyme-altered preneoplastic lesions in rats via angiogenesis suppression. AB - BACKGROUND/AIMS: Chemoprevention should be a promising approach to improve the prognosis of the patients with hepatocellular carcinoma (HCC). Angiogenesis is now recognized as a crucial step not only in tumor growth, but also in early carcinogenesis. The aim of this study was to elucidate the combination effect of the clinically used vitamin K(2) (VK) and the angiotensin-converting enzyme inhibitor, perindopril (PE), on hepatocarcinogenesis, especially in conjunction with angiogenesis. METHODS: In a diethylnitrosamine-induced rat hepatocarcinogenesis model, the effects of VK and PE on the development of liver enzyme-altered preneoplastic lesions and angiogenesis were examined. RESULTS: Treatment with both VK and PE markedly inhibited the development of preneoplastic lesions in association with suppression of neovascularization in the liver. The combination treatment with VK and PE exerted a more potent inhibitory effect as compared with the single agent treatments. The in vitro study demonstrated that VK and PE inhibited the endothelial cell (EC) tubular formation. VK also suppressed the EC proliferation in a dose-dependent manner. CONCLUSIONS: The combination of VK and PE exerted a chemopreventive effect against rat liver carcinogenesis via suppression of angiogenesis. Since both agents are widely used in the clinical practice, this combination therapy may represent a potential new strategy for chemoprevention against HCC in the future. PMID- 15826720 TI - MELD score is a better prognostic model than Child-Turcotte-Pugh score or Discriminant Function score in patients with alcoholic hepatitis. AB - BACKGROUND/AIMS: The aim of the present study was to compare MELD score, Child Turcotte-Pugh (CTP) score, modified Maddrey's Discriminant Function (DF) score, and the related variables in predicting in-hospital mortality of patients with alcoholic hepatitis. METHODS: A retrospective chart review and statistical analyses were done on 202 patients consecutively admitted for alcoholic hepatitis from 1997 to 2002 at the Liver Unit at Rancho Los Amigos Medical Center. RESULTS: Twenty-nine patients died during the hospitalization. Admission MELD score (OR 1.1, P=0.005), first week MELD score (OR 1.2, P<0.0001), and first week increase in MELD score (OR 1.3, P<0.0001) were independently associated with in-hospital mortality. The area under the receiver operating curve (AUC) for the first week increase in MELD score was higher compared to CTP score (P=0.0004) and DF score (P=0.059). Moreover, the first week MELD score >/=20 had the best sensitivity (91%) and specificity (85%) compared with admission or first week change MELD score. CONCLUSIONS: The present study indicates that in patients with alcoholic hepatitis, admission, first week, and first week change in MELD score are significantly independent predictors for in-hospital mortality. MELD score is a more valuable model than CTP or DF score in patients admitted with alcoholic hepatitis. PMID- 15826719 TI - Mild hypothermia prevents brain edema and attenuates up-regulation of the astrocytic benzodiazepine receptor in experimental acute liver failure. AB - BACKGROUND/AIMS: Mild hypothermia has proven useful in the clinical management of patients with acute liver failure. Acute liver failure in experimental animals results in alterations in the expression of genes coding for astrocytic proteins including the "peripheral-type" (astrocytic) benzodiazepine receptor (PTBR), a mitochondrial complex associated with neurosteroid synthesis. To gain further insight into the mechanisms whereby hypothermia attenuates the neurological complications of acute liver failure, we investigated PTBR expression in the brains of hepatic devascularized rats under normothermic (37 degrees C) and hypothermic (35 degrees C) conditions. METHODS: PTBR mRNA was measured using semi quantitative RT-PCR in cerebral cortical extracts and densities of PTBR sites were measured by quantitative receptor autoradiagraphy. Brain pregnenolone content was measured by radioimmunoassay. RESULTS: At coma stages of encephalopathy, animals with acute liver failure manifested a significant increase of PTBR mRNA levels. Brain pregnenolone content and [(3)H]PK 11195 binding site densities were concomitantly increased. Mild hypothermia prevented brain edema and significantly attenuated the increased receptor expression and pregnenolone content. CONCLUSIONS: These findings suggest that an attenuation of PTBR up-regulation resulting in the prevention of increased brain neurosteroid content represents one of the mechanisms by which mild hypothermia exerts its protective effects in ALF. PMID- 15826721 TI - p38 MAP-kinase regulates function of gap and tight junctions during regeneration of rat hepatocytes. AB - BACKGROUND/AIMS: Hepatocyte regeneration is considered to be associated with adaptive changes in expression of gap and tight junctions through multiple signal transduction pathways including p38 MAP-kinase. The role of the stress responsitive MAP-kinase, p38 MAP-kinase, signaling pathway in function of gap and tight junctions was examined during regeneration of rat hepatocytes in vivo and in vitro. METHODS: We examined changes in formation, expression and function of gap and tight junctions in rat livers after 70% partial hepatectomy and in primary cultures of rat hepatocytes, by using a p38 MAP-kinase inhibitor, SB203580. RESULTS: When p38 MAP-kinase was activated during partial hepatectomy, down-regulation of Cx32 and up-regulation of claudin-1 were observed. By SB203580 treatment, the down-regulation of Cx32 was inhibited and the up-regulation of claudin-1 was enhanced, well maintaining the structures of gap and tight junctions. SB203580 treatment did not affect the increase of hepatocyte proliferation. In EGF induced proliferative rat hepatocytes treated with SB203580, the expression and function of Cx32 and claudin-1 were increased. CONCLUSIONS: Dynamic changes of formation of gap and tight junctions during regeneration of rat hepatocytes in vivo and in vitro are in part controlled via a p38 MAP-kinase signaling pathway, and are independent of cell growth. PMID- 15826722 TI - Sexual dimorphism in endotoxin susceptibility after partial hepatectomy in rats. AB - BACKGROUND/AIMS: Liver failure due to endotoxemia after hepatectomy is a fatal complication. Little is known regarding the gender influence on this pathophysiological condition. This study was conducted to investigate whether a gender difference exists in the endotoxin susceptibility after hepatectomy. METHODS: Sexually mature male and female rats received an intravenous administration of lipopolysaccharide (LPS), as endotoxin, 48h after a two-thirds hepatectomy. RESULTS: The 24-h survival rate after LPS administration was significantly higher in females (75%) than in males (38%). Ovariectomy reduced the survival rate in females to 44%. Plasma tumor necrosis factor-alpha levels 1h after LPS were significantly elevated in males and ovariectomized females. The inducible nitric oxide synthase (iNOS) gene expression in liver and spleen, and consequent nitric oxide production 3h after LPS were significantly enhanced in males and ovariectomized females when compared to females, in addition to less functional and structural liver damage in females. CONCLUSIONS: Our results indicate a gender difference in the susceptibility to endotoxemia in the early phase after hepatectomy. Female tolerance to these conditions may be mediated by an inhibition of excessive inflammatory response in the liver and the spleen, partially via the suppression of iNOS gene up-regulation. PMID- 15826723 TI - Systemic efficacy of combined suicide/cytokine gene therapy in a murine model of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Gene therapy might be a promising therapeutic approach for advanced hepatocellular carcinoma (HCC) not amenable to any effective traditional treatment. The aim of the study was to evaluate the in vitro and in vivo efficacy of combined gene therapy of HCC with two different MoMLV-derived retroviral vectors, an MFG- and a LXSN-derived vector, both containing HSV-TK and hIL-2. RESULTS: In vitro experiments on HCC cells showed efficient killing of transduced cells and efficient bystander effect after ganciclovir (GCV) treatment, with higher antitumor activity when the MFG-based vector was used. In vivo studies in a murine syngenic model of HCC demonstrated that treatment with GCV led to complete regression of tumors composed of transduced cells and regression of distant non-transduced tumors. Tumor transduction and efficacy of treatment was also demonstrated after in vivo delivery of vectors. Microarray analysis of tumor samples in mice receiving gene therapy showed up-regulation of genes involved in immune response and signal transduction. CONCLUSIONS: We demonstrated the in vitro and in vivo efficacy of combined retroviral-mediated gene therapy for HCC, with significant systemic therapeutic efficacy in vivo. PMID- 15826724 TI - Increase of doxorubicin sensitivity by doxorubicin-loading into nanoparticles for hepatocellular carcinoma cells in vitro and in vivo. AB - BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is known to be chemoresistant to anticancer drugs due to the multidrug resistant (MDR) transporters expression. Here, we compared in vitro and in vivo the anti-tumor efficacy of doxorubicin loaded polyisohexylcyanoacrylate nanoparticles (PIHCA-Dox) versus free doxorubicin (Dox). These nanoparticles are known to overcome the MDR phenotype. METHODS: We first determined in vitro the 50% inhibition concentration (IC(50)) of these drugs on different human hepatoma cell lines. Secondly, the efficacy of the drugs in vivo was determined on the X/myc transgenic murine model of HCC by histological counting of apoptotic tumorous hepatocytes and by TUNEL labeling. We characterized by semi-quantitative RT-PCR the MDR-related gene (mdr1, mdr3, mrp1) expression pattern in this model. RESULTS: In vitro, IC(50) was reduced with PIHCA-Dox versus Dox for Huh7 (1.7-fold reduction; P<0.001), HepaRG (4.5-fold reduction; P<0.01), HepG2 (1.5-fold reduction; P<0.001), and HepG2.2.15 (1.5-fold reduction; P=0.059). In vivo, HCC in transgenic mice overexpressed the mdr1 and mdr3 genes and the antitumor drugs efficacy was greatly enhanced after injection of PIHCA-Dox (9.0+/-5.0%; n=15) versus Dox (4.6+/-3.3%; n=13; P=0.01) for apoptotic bodies count. CONCLUSIONS: These promising data showing a higher anti tumor efficacy on HCC of PIHCA-Dox versus Dox, warrant further studies in both animals and humans. PMID- 15826725 TI - An in vitro model of hepatitis C virus genotype 3a-associated triglycerides accumulation. AB - BACKGROUND/AIMS: The hepatitis C virus (HCV) induces lipid accumulation in vitro and in vivo. Although clinical observations are consistent with a direct effect of HCV genotype 3a on lipid metabolism, experimental systems have focused on the expression of HCV proteins of genotype 1. To extend these observations, we established an in vitro model expressing the HCV core of different genotypes. METHODS: The HCV core protein from patients with severe (genotype 3a) or no (genotypes 1b, 2a, 3h, 4h and 5a) liver steatosis was expressed in Huh7 cells. Core protein expression (by immunohistochemistry and immunoblot) and triglycerides accumulation (by Oil Red O stain and enzymatic measurement) were evaluated 48h after transfection. RESULTS: Although triglyceride accumulation occurred with genotypes 1b, 3a and 3h, the genotype 3a core protein expression resulted in the highest level of accumulation (i.e. about 3-fold with respect to 1b, and 2-fold with respect to 3h). This effect was not related to core protein expression levels and was abolished by culturing cells in lipid-free medium. CONCLUSIONS: Consistent with observations in chronic hepatitis C patients, the in vitro expression of HCV genotype 3a core protein is the ideal candidate model for studying the mechanisms of HCV-associated steatosis. PMID- 15826726 TI - Hepatic phospholipids in alcoholic liver disease assessed by proton-decoupled 31P magnetic resonance spectroscopy. AB - BACKGROUND/AIMS: Alteration of the phospholipid composition of hepatic biomembranes may be one mechanism of alcoholic liver disease (ALD). We applied proton-decoupled (31)P magnetic resonance spectroscopic imaging ({(1)H}-(31)P MRSI) to 40 patients with ALD and to 13 healthy controls to confirm that metabolic alterations in hepatic phospholipid intermediates could be detected non invasively. METHODS: All patients underwent liver biopsy. Specimens were scored in non-cirrhosis [fatty liver (n=3), alcoholic hepatitis (n=2), fibrosis (n=4), alcoholic hepatitis plus fibrosis (n=16)], and cirrhosis (n=15). {(1)H}-(31)P spectra were collected on a clinical 1.5-Tesla MR system and were evaluated by calculating signal intensity ratios of hepatic phosphomonoester (PME), phosphodiester (PDE), phosphoethanolamine (PE), phosphocholine (PC), glycerophosphorylethanolamine (GPE), and glycerophosphorylcholine (GPC) resonances. RESULTS: The signal intensity ratio GPE/GPC was significantly elevated in cirrhotic (1.19+/-0.22; P=0.002) and non-cirrhotic ALD patients (1.01+/-0.13; P=0.006) compared to healthy controls (0.68+/-0.04), while PE/PC and PME/PDE were significantly elevated in cirrhotic ALD patients compared to controls (1.68+/-0.60 vs. 0.97+/-0.31; P=0.02, and 0.38+/-0.02 vs. 0.25+/-0.01; P=0.002, respectively) and non-cirrhotic patients. CONCLUSIONS: The data support that {(1)H}-(31)P MRSI appears to distinguish cirrhotic from non-cirrhotic ALD patients and confirms changes in hepatic phospholipid metabolism observed in an animal model. PMID- 15826727 TI - The role of hepatitis B virus integrations in the pathogenesis of human hepatocellular carcinoma. PMID- 15826728 TI - Heterozygous G71R-mutation causing Gilbert's syndrome in one of 103 random persons of German descent. PMID- 15826729 TI - Chlamydia pneumoniae infection is an unlikely cause of primary biliary cirrhosis. PMID- 15826730 TI - Growth of giant hepatic hemangioma after triplet pregnancy. PMID- 15826734 TI - Anticonvulsants in bipolar disorder. AB - In recent years, a number of anticonvulsants have been more rigorously investigated for their potential mood-stabilizing properties. They are heterogeneous in their mechanisms of action and in their efficacy in the various mood states in bipolar illness (Table 3). At present, evidence from well controlled studies supports the role of DIV and CBZ in the treatment of acute mania. DIV seems to have better efficacy than lithium in mixed mania or mania associated with depressive symptoms and is recommended as a first-line pharmacologic option in acutely manic or mixed manic patients. Neither CBZ nor DIV have robust evidence supporting their efficacy in the treatment of acute bipolar depression, although DIV clearly possesses beneficial effects on depressive symptomatology and prophylaxis against depressive episodes during long term treatment. Results from a large study indicate that LAM has significant efficacy in bipolar depression without the associated risks of cycle acceleration or manic/hypomanic switches. LAM should be considered a primary option in patients with bipolar depression and in bipolar II patients with rapid cycling. DIV is recommended as a first-line option in bipolar I patients with rapid cycling. LAM has proven efficacy in the prophylaxis of bipolar I disorder and should be considered along with lithium or DIV as treatment of choice in the long term management of bipolar disorder. For the other anticonvulsants, including CBZ and OXC, there is still inadequate evidence of efficacy as monotherapy in the long-term management of bipolar disorder. Even less data exist for other available AEDs, and consensus is growing that someAEDs (eg, GBP) have little or no specific effect in bipolar disorder. Despite the progress made in the past decade, a wider therapeutic armamentarium is critically needed, because a large proportion of bipolar patients do not respond to acute treatments during a manic or depressive episode and have frequent relapse and recurrences during long-term treatment. As additional AEDs become available, rigorously designed and large scale studies examining AEDs as monotherapy and AEDs in combination therapies versus placebo must be undertaken to assess efficacy and safety more adequately to provide better guidance for the clinician faced with the management of this challenging mood disorder. PMID- 15826735 TI - Atypical antipsychotics for bipolar disorder. AB - Atypical antipsychotic agents have been widely investigated for their efficacy in acute mania. The data to date suggest that olanzapine,risperidone, quetiapine, aripiprazole, and ziprasidone are effective, with no significant differences in antimanic efficacy among these agents. These agents are effective as an alternative to lithium or divalproex as monotherapy or in combination with these mood stabilizers. The data concerning their utility in acute bipolar depression and maintenance treatment of bipolar disorder are limited. The studies to date suggest that olanzapine has modest acute antidepressant properties but probably has efficacy comparable to lithium and divalproex in preventing manic and depressive episodes. Quetiapine seems to have robust antidepressant properties, but these data need to be replicated in further trials before quetiapine can be recommended as a first-line agent for acute bipolar depression. Aripiprazole has shown promise in preventing manic episodes in one 6-month study, but further studies with at least 1-year duration and larger sample sizes are needed before this agent can be recommended as a monotherapy for prophylaxis of bipolar disorder. It is currently unknown if risperidone, aripiprazole, and ziprasidone have any efficacy in treating acute bipolar depression. Similarly, long-term studies are needed to ascertain the role of risperidone, quetiapine, and ziprasidone in the maintenance treatment of bipolar disorder. Overall, the atypical antipsychotic agents as a group represent an effective and relatively safe addition to the armamentarium for the treatment of bipolar disorder. PMID- 15826736 TI - Treatment of bipolar depression. AB - This article discusses current practices in the treatment of bipolar depression. In the absence of more definitive research, the treatment of bipolar depression is guided by clinical experience and expert opinion, and sometimes by marketing and popular trends, as much as it is by hard data. Considering the limitations of current knowledge is an essential component of the scientific practice of psychiatry. PMID- 15826737 TI - Psychosocial treatments for bipolar disorders. AB - Psychosocial problems may be causes or consequences of BP relapses,and adding psychologic therapies to usual-treatment approaches may improve the prognosis of those at risk of persistent symptoms or frequent episodes. The three core individual manualized therapies (IPSRT, cognitive therapy, and FFT) have all developed specific models for use in BP. Colom et al's group psychoeducation model also has a clearly developed rationale and format, and it allows individuals to share their views of BP with others, to learn adaptive coping strategies from the other 8 to 12 members of the group, and to have regular contact with an expert therapist. Careful review of the four more extended and comprehensive approaches and the brief technique-driven interventions demonstrates that the effective therapies incorporate one or more of the modules show in Box 1. At present,the choice between the four extended models is more likely to be dictated by patient choice or the availability of a trained therapist. The technique-driven interventions are briefer than the specific therapies (about 6-9 sessions compared with about 20-22 sessions) and usually offer a generic, fixed treatment package targeted at a circumscribed issue such as medication adherence or managing early symptoms of relapse. These brief interventions can be delivered by a less-skilled or less-experienced professional than the specific model. They potentially seem to be useful in day-to-day clinical practice in general adult psychiatry settings; additional larger-scale, randomized trials should be encouraged. Given the reduction in relapse rates and hospitalizations associated with the use of psychologic therapy as an adjunct to medication, it is likely that these approaches will prove to be clinically and cost effective. They may provide a significant improvement in the quality of life of individuals with BP (and indirectly to that of their partners and family members). Brief,evidence-based therapies represent an important component of good clinical practice in the management of BP. Studies of a comprehensive, whole system approach to the collaborative psychobiosocial management of BP are being undertaken in the United States. If these approaches improve the quality and continuity of care for individuals with BP, they will have further implications for the delivery and organization of mental health services. The number and variety of trials of psychosocial interventions is exciting for researchers and clinicians interested in BP. Enthusiasm for advocating these approaches should be tempered by an acknowledgment that the trials undertaken so far largely demonstrate efficacy in selected samples of patients treated at specialist BP clinics or psychologic treatment research centers. Translating efficacy into effectiveness requires evidence that the approaches used in the treatment trials are equally beneficial when used by the wider therapist community treating patients seen routinely in non-specialist or nonresearch centers. These patients often have multiple problems or complex presentations that preclude their involvement in pharmacologic or psychologic treatment studies, but monitoring the outcomes of these representative samples will be important in determining the true place of psychologic approaches in the management of BP. Large-scale studies are now underway on both sides of the Atlantic (the Medical Research Council study in the United Kingdom and the STEP-BD project in the United States). These trials are likely to answer basic questions about the benefits and limitations of psychologic therapies in the acute and maintenance treatment of BP in the clinical realm and will increase understanding of the effectiveness-versus efficacy question. PMID- 15826738 TI - Pharmacotherapy of children and adolescents with bipolar disorder. AB - The identification and treatment of children and adolescents with a bipolar disorder is often challenging and difficult. Many of the psychotropic agents used to treat adults with bipolar disorder may also be-used to treat children and adolescents with these disorders. Further controlled trials using combination pharmacotherapy in children and adolescents with bipolar disorders are needed to advance the field of pediatric bipolarity and provide optimal care for these patients. There are multiple ongoing trials of mood stabilizers and atypical antipsychotics that will provide important controlled data that are currently lacking in the field. PMID- 15826739 TI - Mood-stabilizing drugs: are their neuroprotective aspects clinically relevant? AB - The possibility that there may be subtypes of bipolar disorder and the slow progress in understanding the therapeutic mechanism for approved mood-stabilizing drugs make the challenges of intelligent drug design seem daunting. Nonetheless, the numerous shortcomings in current pharmaco-therapy underscore the need to develop novel therapies. There are significant problems with currently approved mood-stabilizing drugs: 1. Up to 40% of patients fail to respond to monotherapy with either lithium or valproic acid. 2. Common use of polypharmacotherapy increases the side effects associated with treatment. 3. Treatment must continue for weeks to months for therapeutic effects to be greater than placebo. 4. Up to 60% of patients will discontinue therapy, which is somewhat attributable to unwanted side effects. Thus, it is critical that new medications without these problems be developed for bipolar disorder. The hypothesis that mood-stabilizing drugs are neuroprotective is an important first step in new drug development. To determine if the clinical efficacy of mood-stabilizing drugs is dependent on the neuroprotective or neurogenic properties of these medications, greater strides need to be made in relating findings from cell culture and animal models to human imaging and pathology. Mounting evidence supports the neuroprotective and neurogenic properties of lithium and valproic acid ina variety of cell-culture models. It is important for clinical, biochemical, and in vitro differences between these medications to be examined, not ignored,because these differences may reveal critical distinctions between the neural mechanisms of these drugs. Continuation of the in vitro work will aid in the understanding of the mechanism by which these drugs are neuroprotective,but such studies do not advance the understanding of whether these effects are critical for the clinical efficacy of these medications. In attempting to understand the in vivo effects of these medications, a variety of evidence supports the neuroprotective and neurogenic aspects of lithium and valproic acid in healthy rodents and animal models of gross brain insult. More work needs to be done to assess whether these effects occur in animal models for bipolar disorder. The proof of principle for supporting the claim that the neuroprotective or neurogenic properties are important clinically will come from longitudinal clinical studies that compare brain morphology and function before and during treatment. If enough evidence supports the hypothesis that the neuroprotective and neurogenic properties of mood-stabilizing drugs are important for their clinical efficacy, new medications that are more efficacious and have fewer side effects will be designed based on this discovery. PMID- 15826740 TI - Bipolar disorder and substance abuse. AB - SUDs are common in people with BPD. The reasons for this association are not well understood and may be related to several factors. When present,SUDs in BPD patients seem to be associated with a poor prognosis. The treatment of patients with BPD and an SUD has been the subject of relatively little investigation, but medications that are effective mood stabilizers seem to decrease substance use in some reports. PMID- 15826741 TI - Cognition in bipolar disorder. AB - BPD is often associated with cognitive deficits that tend to be present regardless of mood state. Greater impairments tend to be seen in BPD patients who are older, have an early onset of the disease, and suffer a more severe course of illness. The literature also suggests that cognitive deficits are present early in patients with BPD and may be cumulative, showing an association with the number of affective (particularly depressed) episodes over time. Cognitive deficits in BPD may share some common characteristics with those seen in patients with schizophrenia, although the latter tend to show much greater and generalized cognitive impairment. For example, unlike patients with schizophrenia, patients with BPD typically do not score lower than normal persons on measures of global intellectual ability. There also is not overwhelming evidence of laterality or localization of cognitive deficits in BPD, although debate in the literature continues. More visuospatial deficits tend to be found in BPD and UPD than in schizophrenia, thereby raising the possibility of greater involvement of right hemisphere systems in mood disorders. In general, despite variability across investigations, deficits in executive functioning, episodic memory,sustained concentration, and, to a lesser extent, visuospatial skills seem to be the most consistent areas of impairment in BPD. Just as neuroimaging anomalies have been well documented in schizophrenia, structural brain abnormalities have been noted in BPD,most commonly involving the basal ganglia or white matter. Specific comparisons of cerebral atrophy and ventricular size between patients with schizophrenia and BPD have not been definitive, making it difficult to draw conclusions about structural brain abnormalities that might be specific to BPD. Nonetheless, there is enough evidence to suggest that white-matter abnormalities are reported with a greater frequency in BPD patients than in patients with UPD or schizophrenia. Functional neuro-imaging studies of mood disorders have indicated that the frontal cortex,basal ganglia, and temporal lobes are involved. The relationships between neuroimaging and neurocognitive abnormalities in BPD are worthy of additional investigation. Clearly, efforts directed toward phenotyping neuropsychiatric disorders using such measures, in addition to other clinical, neuroimaging, neurophysiologic, and genotypic information, may yield important insights into the development, nature, and course of illness. It is hoped that this understanding will lead to better identification of individuals who may be prone to greater cognitive impairment or decline and those who might be more responsive to specific treatments. PMID- 15826742 TI - Magnetic resonance findings in bipolar disorder. AB - The MR findings reviewed in this article suggest structural, chemical, and functional abnormalities in specific brain regions participating in mood and cognitive regulation, such as the DLPFC, anterior cingulate, amygdala,STG, and corpus callosum in subjects with bipolar disorder. These abnormalities would represent an altered anterior-limbic network disrupting inter- and intrahemispheric communication and underlying the expression of bipolar disorder. Available studies are limited by several confounding variables, such as small and heterogeneous patient samples, differences in clinical and medication status, and cross-sectional design. It is still unclear whether abnormalities in neurodevelopment or neurodegeneration play a major role in the pathophysiology of bipolar disorder. These processes could act together in a unitary model of the disease, with excessive neuronal pruning/apoptosis during childhood and adolescence being responsible for the onset of the disorder and subsequent neurotoxic mechanisms and impaired neuroplasticity and cellular resilience being responsible for further disease progression. Future MR studies should investigate larger samples of first-episode drug-free patients, pediatric patients, subjects at high risk for bipolar disorder, and unaffected family members longitudinally. Such a study population is crucial to examine systematically whether brain changes are present before the appearance of symptoms (eg, maldevelopment) or whether they develop afterwards, as a result of illness course (eg, neurodegeneration). These studies will also be instrumental in minimizing potentially confounding factors commonly found in adult samples, such as the effects of long-term medication, chronicity, and hospitalizations. Juvenile bipolar patients often have a strong family history of bipolar disorder. Future studies could help elucidate the relevance of brain abnormalities as reflections of genetic susceptibility to the disorder. MR studies associated with genetic, post-mortem, and neuropsychologic studies will be valuable in separating state from trait brain abnormalities and in further characterizing the genetic determinants, the neuropathologic underpinnings, and the cognitive disturbances of bipolar disorder. PMID- 15826743 TI - Hypothalamic-pituitary-adrenal axis and bipolar disorder. AB - There is robust evidence demonstrating abnormalities of the HPA axis in bipolar disorder. Hypercortisolism may be central to the pathogenesis of depressive symptoms and cognitive deficits, which may in turn result from neurocytotoxic effects of raised cortisol levels. Manic episodes may be preceded by increased ACTH and cortisol levels, leading to cognitive problems and functional impairments. Identification and effective treatment of mood and cognitive symptoms of mood disorders are clinical goals, but currently available treatments may fall short of this ideal. Manipulation of the HPA axis has been shown to have therapeutic effects in preclinical and clinical studies, and recent data suggest that direct antagonism of GRs maybe a future therapeutic strategy in the treatment of mood disorders. PMID- 15826744 TI - Recent findings on the genetic basis of bipolar disorder. AB - What began as a search for a specific gene for bipolar disorder has now become a search for multiple susceptibility genes as it has be-come clear that the genetic basis of bipolar disorder probably involves multiple genes interacting with each other and with environmental components in as-yet mysterious ways. This article reviews the most recent findings and the emerging picture in the genetics of bipolar disorder. PMID- 15826745 TI - Incidence of prostate cancer in Sicily: results of a multicenter case-findings protocol. AB - OBJECTIVE: To establish the incidence of prostate cancer (PCa) in Sicily in patients who entered an early detection protocol. METHODS: From February 2002 to February 2004, 16,298 subjects aged 40-75 entered the protocol. Patients with suspicious DRE, PSA>10 ng/ml, PSA4 ng/ml underwent a combined MRI/MRSI analysis. Suspicious zones on standard MRI included low intensity signals on T2 weighted images. A high choline+creatine-to-citrate ratio defined a MRSI suspicious zone. A 10 cores following peripheral biopsy scheme was done to which were added supplementary biopsies on the MRI/MRSI suspicious zones. RESULTS: The mean age was 62.3 years (51-74), the mean pre biopsy serum PSA was 12 (3.87-35), the mean free/total PSA ratio was 11% (5-20). The mean number of previous prostate biopsy rounds was 2.04. 15 prostate cancers were diagnosed (35.7%). In 9 cases, abnormal MRI/MRSI findings and positive biopsy sites were located on the same prostatic zones. In 5 cases, MRSI alone located the positive biopsy zones. Sensitivity of combined MRI/MRSI in this study was 73.3%; specificity, positive predictive value, negative predictive value and accuracy were 96.3%, 91.6%, 86.6% and 88% respectively. CONCLUSIONS: This preliminary study shows that the combination of MRI and MRSI might be able to guide and therefore limit the number of iterative biopsies and cores for patients who are at high risk of having a prostate cancer. In some cases, MRSI alone allows identification of neoplasic prostatic zones. Other studies are needed to confirm these data. PMID- 15826748 TI - Efficacy and tolerability of radiotherapy as treatment for bicalutamide-induced gynaecomastia and breast pain in prostate cancer. AB - OBJECTIVE: To assess the efficacy and tolerability of localised radiotherapy for the treatment of bicalutamide ('Casodex''Casodex' is a trademark of the AstraZeneca group of companies.)-induced gynaecomastia and/or breast pain. METHODS: This open-label, non-comparative, multicentre study included 51 patients receiving bicalutamide 150 mg for the treatment of non-metastatic prostate cancer (T1b-T4, Nx, M0). Patients who developed symptomatic gynaecomastia and/or breast pain received two 6-Gy fractions of external-beam radiation to the breasts and were then assessed at two 3-monthly follow-up visits. RESULTS: 37/51 (72.5%) patients experienced gynaecomastia and 41/51 (80.4%) experienced breast pain, typically within the first 6 months. Twenty seven and 38 patients, respectively, went on to receive breast irradiation. Following radiotherapy, gynaecomastia improved or resolved in 7/27 (25.9%) and 2/27 (7.4%) cases, respectively, and breast pain improved or resolved in 12/38 (31.6%) and 3/38 (7.9%) cases, respectively. No change was observed in 7 patients (25.9%) with gynaecomastia and 12 patients (31.6%) with breast pain, while 9 patients (33.3%) and 8 patients (21.1%), respectively, worsened. Radiotherapy-related adverse events, reported by 18/41 (43.9%) patients, were generally mild and short lived (median duration approximately 5 weeks). CONCLUSIONS: Therapeutic radiotherapy, using two fractions of 6 Gy external-beam radiation to the male breast, improves the intensity of bicalutamide-induced gynaecomastia and/or breast pain in approximately one-third of patients. Adverse events were often mild and short lived. PMID- 15826749 TI - Expression of the endothelin axis in bladder cancer: relationship to clinicopathologic parameters and long-term survival. AB - OBJECTIVES: Endothelin-1 (ET-1) and its receptors ET(A)R and ET(B)R, referred to as the Endothelin-axis, play an emerging role in cancer. We examined the ET-axis immunohistochemically in invasive bladder cancer. METHODS: Tumor specimens from 157 patients after cystectomy were stained immunohistochemically for ET-1, ET(A)R and ET(B)R. After semiquantitative analysis the staining results were correlated with clinicopathological parameters and survival rates. RESULTS: Overexpression of ET-1, ET(A)R and ET(B)R was identified in 26.8%, 58.8% and 76.9% of cases, respectively. No association with TNM staging and histologic grading was found. However, patients with ET(B)R expression tended to have organ-confined tumors (p=0.16) and no vascular invasion (p=0.09), the latter being statistically significant in the subgroup of G3 tumors (p=0.02). ET(B)R overexpression was associated with favorable disease-free survival (p=0.04). CONCLUSIONS: The ET axis is overexpressed in bladder cancer, ET(B)R predominating in this entity and being associated with a more favorable prognosis. Further studies are warranted to elucidate the role of the ET-axis as a molecular target in bladder cancer. PMID- 15826750 TI - Dynamic sentinel node biopsy in penile carcinoma: evaluation of 10 years experience. AB - PURPOSE: The aim of this study was to evaluate the results of 10 years dynamic sentinel node biopsy experience in penile carcinoma at our institute. PATIENTS AND METHODS: 140 patients with clinically node-negative groins were prospectively included. Lymphoscintigraphy was performed after injection of 99mTechnetium nanocolloid around the primary tumour. The sentinel node was intraoperatively identified with the aid of patent blue dye and a gamma ray detection probe. Lymph node dissection was performed only if sentinel node metastasis was found. Median follow-up was 52 months (range 5-129). RESULTS: Lymphoscintigraphy visualized at least 1 sentinel node in 138 patients. Sentinel node metastasis was found in 37 inguinal regions of 31 patients. The sentinel node was the only tumour-positive node in 78% (29/37) of the dissection specimens. Complications occurred in 8% (17/206) of the operated groins. False-negative results were encountered in 6 patients resulting in a false-negative rate of 16% (6/37 patients). 5-year disease-specific survival was 96% and 66% for patients with a tumour-negative sentinel node and tumour-positive sentinel node, respectively (p=0.001). CONCLUSION: Dynamic sentinel node biopsy in penile carcinoma is of important diagnostic, prognostic, and therapeutic value at the cost of only minor morbidity. PMID- 15826751 TI - The effect of biofeedback physical therapy in men with Chronic Pelvic Pain Syndrome Type III. AB - Recent studies suggest that the symptoms of chronic non-bacterial prostatitis (CP) or Chronic Pelvic Pain Syndrome (CPPS) may be due to or associated with pelvic floor muscle dysfunction. Therapies aimed to improve relaxation and proper use of the pelvic floor muscles such as biofeedback physical therapy and pelvic floor re-education are expected to give symptom improvement. The objective of this study was to evaluate the effect of biofeedback physical therapy on the symptoms of men with CPPS. MATERIALS AND METHODS: Between March 2000 to March 2004, 33 consecutive men were diagnosed with CP/CPPS based on history including the NIH-CPSI questionnaire and physical examination including pelvic floor muscle tonus, urinalysis, uroflowmetry with residual urine measurement and transrectal ultrasonography of the prostate. All patients participated in a pelvic floor biofeedback re-educating program. A rectal EMG probe was used to measure resting tone of the pelvic floor muscles and was helpful for instruction pelvic floor muscles contraction and relaxation. RESULTS: Two of the 33 men dropped out. In the remaining 31 men, mean age 43.9 years (range 23-70), the mean total Chronic Prostatitis Symptom Index (NIH-CPSI) changed from 23.6 (range 11-34) at baseline to 11.4 (range 1-25) after treatment (p<0.001). The mean value of the pelvic floor muscle tonus was 4.9 at diagnosis (range 2.0-10.0) and decreased to 1.7 (range 0.5-2.8) after treatment (p<0.001). CONCLUSIONS: Our study clearly demonstrates a significant effect of biofeedback physical therapy and pelvic floor re-education for CP/CPPS patients, leading to a significant improvement of the symptom score. The correlation between the pelvic muscle tonus results with NIH-CPSI score is highly suggestive that the pelvic floor plays an important role in the pathophysiology of CP/CPPS. PMID- 15826752 TI - Vardenafil for the treatment of erectile dysfunction: a critical review of the literature based on personal clinical experience. AB - OBJECTIVE: To critically review the literature on vardenafil in the treatment of erectile dysfunction while integrating the clinical findings with the personal experience of the authors. METHODS: Analysis of published full-length papers that were identified through Medline search from January 2000 through May 2004. Abstracts published in peer-reviewed journals from the same period were also considered. RESULTS: Efficacy, tolerability and safety, as reported in the peer reviewed literature compares well with the authors' personal experience. Authors' personal observations include discussions on potency, selectivity, selection of initial dose, counselling for patients characteristically considered difficult-to treat (diabetes, prostatectomy, depression), including the determination of the maximal efficacious dose and the possible role of daily dosing, optimisation of the use of vardenafil according to its pharmacokinetic and pharmacodynamic profiles (onset and reliability), and management of ED patients with or at risk for cardiovascular disease. CONCLUSIONS: Extensive experience with vardenafil as reported in peer reviewed literature confirms the important role of vardenafil in the management of patients with ED. The development of each physician's own experience with vardenafil is key to optimise overall satisfaction of this therapy by the patient and his partner. PMID- 15826753 TI - Laparoscopic adrenal surgery for recurrent tumours in patients with hereditary phaeochromocytoma. AB - OBJECTIVE: To report our experience with Laparoscopic Partial Adrenalectomy (LPA) for recurrent tumours in patients with hereditary phaeochromocytoma. PATIENTS AND METHODS: Five patients with hereditary phaeochromocytoma (4 with von Hippel Lindau disease and 1 with Multiple Endocrine Neoplasia 2B), who had undergone adrenal surgery previously, presented with recurrent adrenal tumours. One patient was pregnant at 20 weeks of gestation. All patients underwent hormonal evaluation, genetic screening and imaging with CT or MRI, metaiodobenzylguanidine (MIBG) scintigraphy. RESULTS: Of the 7 attempted LPA in five patients, five procedures (71%) were successfully completed and total adrenalectomy was needed on two occasions. The adrenal vein could be spared in all patients except one. There were no intra-operative complications. The adrenal function was adequate in all patients without need for steroid supplementation except one patient who lost both adrenals eventually. There was no correlation between the preservation of adrenal vein and adrenocortical function. CONCLUSION: Laparoscopic partial adrenalectomy is feasible, safe and effective in recurrent phaeochromocytoma, despite previous adrenal surgery and is technically easier if the previous approach had been laparoscopic as well. Patients with hereditary phaeochromocytoma are prone for recurrent tumours and may need repeated surgical procedures. Hence, minimally invasive approach is ideal for these patients. PMID- 15826754 TI - Innervation of the female human urethral sphincter: 3D reconstruction of immunohistochemical studies in the fetus. AB - OBJECTIVES: The precise location, origin and nature of nerve fibers innervating the urethral sphincter have not been clearly established. Classical anatomical studies based on cadaver dissections have provided conflicting results concerning the location of pudendal and autonomic nerve fibers. This study was designed to identify nerve fibers innervating the urethral sphincter and to provide a three dimensional representation of their tissue relations in the female human fetus. MATERIALS AND METHODS: Histology and immunohistochemistry (Masson's Trichromic, Luxol Fast Blue, Protein S 100 immunostaining and smooth fiber actin immunostaining) were performed on the external urethral sphincter of ten female fetuses with a crown-rump length of 112 to 340mm. Three-dimensional reconstructions of the urethral structure and innervation were obtained from serial sections using Surf Driver 3.5.3 software (David Moody and Scott Lozanoff). RESULTS: Three-dimensional reconstructions of the same sections with different stains demonstrated the precise structure of the muscle layers (smooth and striated muscle fibers) and nerve fibers (myelinated and unmyelinated) and their relations with the urethra and vaginal wall. The proximal third consisted of a circular smooth muscle sphincter, the middle third consisted of two circular layers of smooth and striated muscle fibers and the distal third consisted of a circular layer of smooth muscle fibers surrounded by an omega-shaped layer of striated muscle fibers. In the proximal third of the urethral sphincter, myelinated fibers were identified running with unmyelinated fibers from the pelvic plexus. These fibers were closely related to the lateral and anterior aspects of the vagina. Unmyelinated fibers entered the smooth muscle part of the sphincter at 4 o'clock and at 8 o'clock. Most myelinated fibers entered the sphincter at 3 o'clock and at 9 o'clock. CONCLUSION: Histological and immunohistochemical three-dimensional reconstruction of the anatomical structures of the urethral sphincter provides a better understanding of the origin and nature of the Innervation participating in urinary continence. It provides a very informative view of the three-dimensional arrangement of sphincter muscle layers. PMID- 15826755 TI - Outcome following TVT sling procedure: a comparison of outcome recorded by surgeons to that reported by their patients at a London district general hospital. AB - OBJECTIVE: Retrospective study to assess patient satisfaction rates after TVT sling procedure for stress urinary incontinence (SUI), and comparison of these results to the post-operative progress documented by the surgeons in the clinical notes. METHODS: All TVT sling cases at our institution during February 1999 to December 2002 were included. Data was collected from clinical notes on post operative outcome as recorded by the surgeons, and a patient satisfaction questionnaire was used to assess patients' perception of their progress. RESULTS: The response rate to the patient satisfaction questionnaire was 72%. Overall 94% of the patients were satisfied with the procedure. However, they reported cure rates of only 44%. This is comparable to the 46% cure rates documented by the surgeons. CONCLUSION: The surgeons' and the patients' perceptions regarding outcome and change in symptoms after TVT sling for SUI at our hospital, correlate well. Hence, the documentation in the clinical notes by the surgeons can be relied upon as an accurate representation of post-operative patient progress. Despite the relatively low cure rates (44-46%) for SUI with TVT sling procedure, the patient satisfaction rates with the outcome are high (94%), reiterating that there is a poor correlation between quality of life impairment and the concept of cure. PMID- 15826756 TI - Erosions and urinary retention following polypropylene synthetic sling: Australasian survey. AB - INTRODUCTION: There are few published reports on the incidence and management of urethral and vaginal erosions following the use of polypropylene synthetic slings. Moreover, there is very little Australasian data on their use for management of female urinary incontinence or on their associated complications. METHODS: A 1-page survey was mailed to the Australian and New Zealand members of the Urological Society of Australasia (N=326). The survey included questions on the use of polypropylene synthetic sling and the incidence and management of post operative vaginal and urethral erosions and urinary retention. RESULTS: 198 surveys were returned (response rate: 61%). Polypropylene synthetic sling procedure is practiced by 39% of the respondents with a total of 1459 cases (TVT: 993, SPARC: 466). The incidence of vaginal erosions, urethral erosions and urinary retention was 1.2%, 0.6% and 6.5%, respectively. Thirty five percent of vaginal erosions were asymptomatic and only identified on routine post-operative vaginal examination. One third of urethral erosions presented more than 1 year after surgery and 89% of these were symptomatic. Thirty four percent of patients with urinary retention required surgical intervention to correct the retention. CONCLUSIONS: The incidence of urethral and vaginal erosions following polypropylene synthetic sling procedures is lower than that with other synthetic slings. However, a high index of suspicion and long-term follow-up are required to identify and manage these complications of this relatively new procedure. PMID- 15826757 TI - Surgical management of stress urinary incontinence: a questionnaire based survey. AB - OBJECTIVES: To determine the trends in the surgical management of urinary stress incontinence amongst members of the International Urogynaecology Association (IUGA). DESIGN: Postal Questionnaire Survey. METHODS: 530 members of the International Urogynaecology association were sent a postal questionnaire regarding their practice in the surgical management of urinary stress incontinence. We also collected data on the demographic profile of members and the preferred primary and secondary continence procedures. OUTCOME: Consensus in the surgical management of urinary stress incontinence amongst members of the IUGA. RESULTS: Five hundred and thirty questionnaires were sent to IUGA members worldwide. Two hundred and seventeen questionnaires were received of which fifteen were from physiotherapists, so were excluded from our analysis. The overall response rate was 41%, and the useable response rate was 38%. Of the total results analysed (two hundred and two), one hundred and twelve (54%) were from teaching hospital, sixty-two (31%) were from district general hospitals and twenty-eight (14%) were from members in private practice. The preferred primary continence procedure was Tension Free Vaginal Tape (TVT) in one hundred and thirty four (68%) respondents. The preferred secondary continence procedure was colposuspension or Trans obturator tape in twenty-six respondents each (13%). CONCLUSIONS: Almost all respondents (97.1%) were skilled at performing either TVT or colposuspension, which have been identified as the preferred methods of surgical management by the NICE (National Institute of Clinical Excellence, UK). Although colposuspension has been identified as the gold standard surgical procedure in the management of stress incontinence, 16% of respondents were not performing colposuspension. There appears to be little evidence base to the surgical techniques in the management of stress urinary continence. PMID- 15826758 TI - Success of repeat detrusor injections of botulinum a toxin in patients with severe neurogenic detrusor overactivity and incontinence. AB - OBJECTIVES: Detrusor injections with botulinum toxin type A are an effective treatment for neurogenic detrusor overactivity, lasting for 9-12 months. When the patients develop botulinum resistance, subsequent injections might be less effective. Repeat injections in patients with severe neurogenic detrusor overactivity and incontinence were studied. METHODS: Patients received Botox (300 UI) or Dysport (750 UI) injections. Clinical variables: satisfaction, anticholinergics use, mean and maximum bladder capacity, continence volume. Cystometric parameters: compliance, cystometric capacity, reflex volume. STATISTICS: Anova, chi2-tests; t-tests and paired t-tests (p=0.05). RESULTS: Forty-three men and 23 women (mean age 38.3 years; mean duration of lesion 9.2 years) were included. The interval between subsequent injections (on average 9-11 months) did not change significantly (p=0.5594). The satisfaction was high and anticholinergics use decreased substantially (p=0.0000). Significant improvements were found in clinical parameters and in cystometric capacity, for compliance only at the second treatment. The incidence of reflex contractions was significantly reduced. Four patients had transient adverse events after Dysport. CONCLUSIONS: Repeat injections with botulinum toxin type A are as effective as the first one. The cause for repeat treatment is relapse of overactive bladder symptoms. PMID- 15826759 TI - Prolonged sacral neuromodulation testing using permanent leads: a more reliable patient selection method? AB - OBJECTIVE: To assess the effect of prolonged sacral neuromodulation testing using permanent leads comparing the usual evaluation period of 4 to 7 days to a prolonged evaluation period of a minimum of 14 days. PATIENTS AND METHODS: A consecutive series of 20 patients (16 females and 4 males) undergoing prolonged sacral neuromodulation testing using permanent leads between September 2000 and March 2004 were evaluated retrospectively. 10 suffered from urgency-frequency syndrome, 3 from urge incontinence and 7 from non-obstructive chronic urinary retention. Key bladder diary variables at baseline, after the usual and prolonged evaluation period and at the last follow-up were compared. RESULTS: The median age was 52 years (interquartile range (IQR) 38-59) and the median evaluation period 28 days (IQR 18-29). 16 of the 20 patients (80%) had successful prolonged sacral neuromodulation testing and underwent the implantation of the IPG that was placed in the anterior abdominal wall in 6 and in the upper buttock in 10 patients. The eligibility for IPG implantation was significantly (p=0.031) increased from 50% after the usual to 80% after the prolonged evaluation period. At a median follow-up of 22 months (IQR 12-34), sacral neuromodulation was successful in 14 (88%) of the 16 IPG implanted patients but failed in 2. CONCLUSIONS: Prolonged sacral neuromodulation testing using permanent leads is more reliable for accurate patient selection than the usual evaluation period. Therefore, this method is strongly recommended and suggested to become the standard test procedure. PMID- 15826760 TI - Clinical experience with the N-shaped ileal neobladder: assessment of complications, voiding patterns, and quality of life in our series of 58 patients. AB - PURPOSE: The purpose of this retrospective study was to assess complications, voiding patterns, and quality of life in patients with an orthotopic bladder substitution, using an N-shaped ileal neobladder. MATERIALS AND METHODS: Between May 1996 and December 2002, 58 patients (52 men and 6 women) underwent an orthotopic ileal neobladder reconstruction after radical cystectomy. The mean age was 47 for the female and 60 for the male patients. In all patients an N-shaped ileal pouch was constructed. This pouch has not yet been described in the literature before. All procedures were performed by the same surgeon (HVP) and the mean follow-up was 38 months. Complications were registered as early (occurring within 3 months) or late (occurring after 3 months), and as pouch related and non-pouch-related. The patients took part in a pelvic floor re education programme for as long as they were incontinent. All patients completed a retrospective Quality of Life questionnaire, based on the QLQ-C30 questionnaire, which was validated by the EORTC's Study Group on Quality of Life. RESULTS: In 38% of the patients, early complications occurred, whereas 48% had late complications. The most frequent early complications were diarrhea (24%) and pyelonephritis (9%). Diarrhea was again the most frequently mentioned non-pouch related complication (19%). The most frequently observed pouch-related late complication was ileo-urethral stenosis. This occurred in five patients. All of these 5 patients were re-operated using a minimally invasive approach. Daytime continence was achieved in 95% of patients and nighttime continence in 66%. Hyper continence with subsequent need for CISC was observed in 5 out of 6 women (83%) and 0 out of 52 men (0%). The retrospective QoL questionnaire learned that the impact of bladder removal and orthotopic bladder substitution has acceptable impact on patient's everyday life. Diarrhea was mentioned as being the most discomforting complication by most of the patients. CONCLUSIONS: We describe a modified orthotopic ileal neobladder: the ileal N-pouch. The functional results with this pouch are good. Complication rates and QoL are comparable with the larger series published by other authors, using different ileal neobladder reconstructions. PMID- 15826761 TI - Prevention of chronic urinary retention in orthotopic bladder replacement in the female. AB - OBJECTIVE: Chronic urinary retention is a frequent complication after orthotopic bladder replacement. Herein a new technical modification to avoid voiding dysfunction is described. METHODS: Between January 1995 and January 2004, 62 women had orthotopic bladder replacement after radical cystectomy. From November 1998 on, 35 patients were operated using a new technical modification, consisting in the fixation of a large and thick flap of peritoneal and extraperitoneal tissue deriving from the posterior wall of the vagina to the edges of endopelvic fascia to create a resistant hammock under the reservoir. RESULTS: 25 women have a follow-up longer than 12 months, range 12-56, and are considered in this study. Only one, out of 25 patients, still has occasionally stress incontinence but she did not need to use pads. Two patients are incontinent at night. None of 25 patients has a residual urine volume greater than 100 ml or needed intermittent catheterization. Median bladder capacity is 350 ml, range 280-430 ml. Median creatinine level is 1.1 mg/dl, range 0.7-1.2 mg/dl. CONCLUSION: The creation of a posterior support to the neobladder, by harvesting a thick flap, has, in our series, completely avoided the occurrence of chronic urinary retention, which is the most common long-term complication of bladder replacement in the female. Our series is sufficiently large to exclude serendipity and to be compared with other series in the literature. PMID- 15826762 TI - Comparison of orthotopic sigmoid and ileal neobladders: continence and urodynamic parameters. AB - OBJECTIVES: The objective this of the study was to compare continence rates and urodynamic parameters among patients who had undergone orthotopic bladder substitution with sigmoid or ileal segments. METHODS: Continent urinary reservoirs were constructed in 112 patients. Fifty patients received a sigmoid neobladder (SN) and 62 patients an ileal neobladder (IN). Thirty-four patients with an SN (mean age 64.4 years), and 20 with an IN (mean age 57.8 years) agreed to postoperative urodynamic evaluation at a median time after surgery of 18 and 37 months, respectively. Continence and urodynamic parameters were compared in both groups. RESULTS: The average reservoir capacity of the SN (296 ml) was lower than the IN (546 ml). The majority of patients voided by the Valsalva maneuver and achieved good peak flow rates [SN group 16.6 (range 7-32) ml/s, IN group 25.5 (range 5-35) ml/s]. Of the patients with an SN 26 (76%) and with an IN 15 (75%) emptied to near completion with a post-void residual (PVR) of less than 100 ml. Daytime continence was achieved in 90% of IN patients and 85% of SN patients. Only 9% of patients with an SN and 60% of patients with an IN were continent at night. CONCLUSION: A neobladder constructed from detubularized ileum or sigmoid achieves adequate capacity with a satisfactory daytime continence rate. Nighttime incontinence in patients with IN can be at least partly explained by periods of high pressure due to neobladder contractions in combination with a relaxed sphincter during sleep. The low nighttime continence rate of the SN is probably related to its small functional capacity. PMID- 15826763 TI - How to interpret a deterioration of split function? AB - OBJECTIVE: A drop of split renal function often constitutes a criterion for pyeloplasty in hydronephrosis since it is considered as representing deterioration of the affected kidney. The aim of this work was to determine, in a selected population of patients with a drop of split function of at least 5%, if the evolution of single kidney glomerular filtration rate (SKGFR) was parallel to the one of split renal function. METHODS: From a large data basis, we found retrospectively only 29 children (10 below and 19 above two years of age at first examination) having had at least two Tc-99m mertiatide (Tc-99m MAG3) renographic explorations for various urological diseases, with a decrease of split function of at least 5% between the two examinations. Evolution of split function was compared to evolution of SKGFR obtained by means of the combination of Tc-99m MAG3 split function and overall glomerular filtration rate as given by the chromium Cr 51 ethylenediamine tetraacetic acid (EDTA) clearance. RESULTS: For the group above two years of age, SKGFR increased or remained stable in 63% of the cases, while in the children less than 2 years of age, a decrease of SKGFR was never observed, according to the maturation of overall GFR in this age group. Thus, the decrease of split function was not necessarily associated with a similar decrease of SKGFR. CONCLUSION: In patients with unilateral or bilateral urological disorders, deterioration of split renal function does not necessary correspond to a loss of function of the affected kidney. SKGFR often modifies the interpretation of split function. PMID- 15826764 TI - Complete repair of bladder exstrophy in boys: can hypospadias be avoided? AB - INTRODUCTION: Complete repair of classic bladder exstrophy in male newborns has been successful with minimal morbidity. However, the technique may create hypospadias in some cases. We have recently adopted a modification to obtain an orthotopic meatus in bladder exstrophy boys. MATERIALS AND METHODS: Between November 1998 and December 2002 complete repair of classic bladder exstrophy was carried out in 27 boys. Complete penile disassembly was performed in 22 boys including 4 newborns and 18 older children; mean age 3+/-2 years old. Modified disassembly was used in the last 5 boys including 4 newborns and a 9-month-old boy. During repair of epispadias, the dissection starts on the ventral aspect of the penis as usual. The urethral plate is separated from both corpora cavernosa to allow ventral transposition of the plate. In the complete disassembly technique, the urethral plate is completely separated from both hemiglans. In this modification, while the urethral plate is completely separated from both corpora cavernosa, the extreme distal end of the urethral plate remains attached to the distal ends of both hemiglans. Thus, when the plate is tubularized with fine interrupted sutures the meatus ends up at the tip of the glans penis. The symphysis pubis is re-approximated as usual and corpora cavernosa are approximated dorsally in the midline. RESULTS: There was no major complication. Mean follow-up was 43+/-7 and 7+/-4 months for complete and modified disassembly groups respectively. Of the 22 boys, who underwent complete disassembly, 15 (68%) ended up with hypospadias and 7 (32%) had an orthotopic meatus. Modified disassembly has resulted in an excellent cosmetic appearance and orthotopic meatus in the 5 boys. Penile length was not shorter than boys who underwent complete disassembly. Parents of the 5 boys noticed normal straight morning erection. CONCLUSION: Although complete penile disassembly allows ventral placement of the urethra, hypospadias is created in approximately two thirds of the cases. When the extreme distal end of the urethral plate remains attached to the distal ends of both hemiglans (modified disassembly), orthotopic meatus can be obtained in all boys. With modified disassembly, posterior mobilization of the bladder and urethra does not result in corporal angulations or shortening because of the proportionate inward movement of the corpora that accompanies symphyseal approximation. The modification is feasible in newborns and infants. These short term results may obviate the need for later penile reconstructive procedure. PMID- 15826765 TI - Insulin-like growth factor binding protein 2: an androgen-dependent predictor of prostate cancer survival. AB - BACKGROUND: Evidence suggests that the insulin-like growth factor (IGF) family is important in prostate cancer. We evaluate the ability of IGF markers to predict biochemical recurrence-free survival (bRFS) following radical prostatectomy (RRP). METHODS: Preoperative sera from 141 patients undergoing RRP were analyzed for IGF-I, IGFBP-1, IGFBP-2 and IGFBP-3. A multivariate Cox model was created to assess the ability of these markers to predict bRFS. Preoperative covariables included: biopsy Gleason score, clinical TNM stage, serum PSA and neoadjuvant hormonotherapy. Kaplan-Meier curves were stratifying by IGF cutpoints (determined by ROC analysis) and hormonotherapy status. RESULTS: Average follow-up was 6.92 years, median PSA was 6.9 ng/ml and 85.1% of patients had cT2NxM0 disease. 49 patients experienced a PSA failure. Average levels of IGF-I, IGFBP-1, IGFBP-2 and IGFBP-3 were 156.5 ng/ml, 48.1 ng/ml, 396.8 ng/ml and 3303.7 ng/ml, respectively. IGFBP-2 was an independent predictor of PSA failure. Patients treated with neoadjuvant ADT (n=71) and whom had high IGFBP-2 levels experienced a 5-year bRFS that was better than those with low IGFBP-2 levels (77.7% vs. 53.3%). Patients without neoadjuvant ADT whom had high IGFBP-2 levels had a 5-year bRFS that was worse than those with low IGFBP-2 levels (64.5% vs. 82.7%). CONCLUSIONS: Preoperative IGFBP-2 predicts post-radical prostatectomy bRFS and is independent of stage, Gleason score and PSA. Increased IGFBP-2 is associated with better survival in patients with neoadjuvant hormonotherapy but worse survival in those without, suggesting a dramatic switch in function of this protein dependent on the presence or absence of androgens. PMID- 15826766 TI - Inhibition of bcl-2 enhances the efficacy of chemotherapy in renal cell carcinoma. AB - OBJECTIVES: Renal cell cancer (RCC) is highly resistant to chemotherapy. Increased expression of the antiapoptotic gene bcl-2 in tumors is known to be associated with poor responses to systemic treatment of cancer. Down-regulation of bcl-2 expression using antisense oligonucleotides (asON) has been shown to increase chemosensitivity in clinical phase I-III studies with various cancers. However, no studies on the efficacy of this approach in RCC have been reported so far. This study aimed to evaluate whether bcl-2 asON could enhance efficacy of chemotherapy in human RCC. MATERIAL AND METHODS: Expression of bcl-2 mRNA and protein was analyzed in different RCC cell lines by RT-PCR and Western blot. Cells with high or low bcl-2 mRNA and protein expression were treated with different concentrations of bcl-2 asON in combination with cisplatin. AsON induced down-regulation of bcl-2 mRNA and protein was documented by RT-PCR and Western blot. Treatment effects on cell viability were analyzed by colorimetric tetrazolium (MTT) assay. Immunohistochemical staining of M30-positive cells was performed for quantification of apoptotic cells. RESULTS: Transfection of high bcl-2 expressing cells with bcl-2 asON alone induced no reduction of cell viability at a concentration range from 100-1000 nM. In combination therapy, pretreatment with asON significantly enhanced MTT reduction after cisplatin treatment. IC50 concentrations of cisplatin were 1 microg/ml with and 2.7 microg/ml without prior incubation. The marked reduction of cell viability correlated with an 8-fold increase of apoptotic cells after combination treatment. Only a minor increase of cisplatin effectivity was noted after asON preincubation of cells with lower bcl-2 expression. CONCLUSIONS: The combination of cisplatin and bcl-2 antisense ON exerts significantly greater effects on cell viability and apoptosis than either agent used alone on human RCC cells. These data indicate that inhibition of bcl-2 expression may be an attractive therapeutic strategy in RCC tumors with high bcl-2 expression. PMID- 15826767 TI - Isolation of primary endothelial and stromal cell cultures of the corpus cavernosum penis for basic research and tissue engineering. AB - OBJECTIVES: Primary cell cultures derived from the corpus cavernosum are frequently used as in vitro models to define cellular mechanisms involved in erectile function. However, previous studies often lack detailed isolation protocols or a precise characterisation of the culture composition excluding especially contaminating fibroblasts. This study aimed at critically analysing and reproducing reported isolation methods, as well as establishing new procedures to receive highly pure and morphologically differentiated endothelial, smooth muscle and fibroblastic cells derived from the human penis. METHODS: We evaluated numerous isolation and enrichment techniques using cavernosal tissue from 57 patients. Assessment factors displayed the purity, cell yield, practicability and reproducibility. The purity in cultured cells was analysed using immunocytochemistry and Western blots. RESULTS: An enzymatic protocol was established for the isolation and cultivation of cavernosal endothelial cells with an impressive purity of 98.0+/-0.8%. In contrast, already published nearly pure smooth muscle cell cultures were not reproducible in our laboratory. Meaningful evidence for an overwhelming presence of fibroblasts in these widely accepted pure smooth muscle cell cultures is presented. CONCLUSION: Endothelial cell cultures derived from human corpora cavernosa are reproducible and reliable to serve for cell culture-based investigations of the endothelial dysfunction. The discrepancy in the purity of smooth muscle cell cultures might reflect laboratory and tissue source factors, lacking an exclusion of fibroblasts in other studies or changes in stromal phenotype under culture conditions. Further research is necessary to clarify a possible plasticity between smooth muscle cells and (myo)fibroblasts and assess functional properties. PMID- 15826768 TI - Re: Stancik I, Luftenegger W, Klimpfinger M, Muller MM, Hoeltl W. Effect of NIH IV prostatitis on free and free-to-total PSA. Eur Urol 2004;46:760-4. PMID- 15826769 TI - Re: Granitsiotis P, Kirk D. Chronic testicular pain: an overview. Eur Urol 2004;45:430-6. PMID- 15826771 TI - Regulation of uncoupling proteins 2 and 3 in porcine adipose tissue. AB - This study was performed to determine whether or not uncoupling protein 2 (UCP2) and UCP3 expression in porcine subcutaneous adipose tissue are hormonally regulated in vitro and whether their expression is correlated with changes in metabolic activity. Tissue slices (approximately 100 mg) were placed in 12-well plates containing 1 mL of DMEM/F12 with 25 mM Hepes, 0.5% BSA, pH 7.4. Triplicate slices were incubated with basal medium or hormone supplemented media at 37 degrees C with 95% air/5% CO2. Parallel cultures were maintained for either 2 or 24 h to evaluate metabolic viability of the tissue. Slices were transferred to test tubes containing 1 mL of DMEM/F12 with 25 mM Hepes, 3% BSA, 5.5 mM glucose, 1 microCi 14C-U-glucose/mL and incubated for an additional 2 h at 37 degrees C. Glucose metabolism in 2-h incubations did not differ from 24-h (chronic) incubations, indicating viability was maintained (P>0.05). Expression of UCP2 and UCP3 was assessed in slices following 24h of incubation with various combinations of hormones by semi-quantitative RT-PCR. Expression of UCP2 was induced by leptin (100 ng/mL; P<0.05). Growth hormone (100 ng/mL) inhibited UCP2 expression (P<0.05). Expression of UCP3 was inhibited by growth hormone (100 ng/mL; P<0.05), tri-iodothyronine (10 nM; P<0.05) or leptin (100 ng/mL; P<0.05). Changes in UCP expression could not be associated with overall changes in glucose metabolism by adipose tissue slices in chronic culture. PMID- 15826772 TI - Evidence for a functional bone morphogenetic protein (BMP) system in the porcine ovary. AB - Bone morphogenetic proteins (BMPs) play important roles in controlling fertility and ovulation rate. There is however, little information on the BMP system in the ovary of a large polyovular species. The aims of the present study were to investigate BMP-2 and -6 protein expression in the porcine ovary, their effects on granulosa cells in culture and their mechanism of action. Cells and oocytes were recovered from healthy antral follicles 2-6mm in diameter. When assessed by Western blotting, oocytes and follicular fluid contained BMP-2 and -6. In addition, BMP-2 and -6 were observed in granulosa cells and BMP-2 was also found in theca cells. Granulosa cells were cultured in a serum-free system for 144 h in the presence of increasing doses (0, 3, 30 and 100 ng/ml) of BMP-2 or BMP-6. Both BMPs suppressed progesterone production in a dose-dependent manner after 48 h (P<0.001) and 144 h (P<0.05). Only BMP-6 stimulated cell proliferation at 100 ng/ml (P<0.05). Investigation into the mechanism of action found that BMP-2 and 6 decreased cyclic adenosine monophosphate (cAMP) production (P<0.01), expression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) protein (P<0.001) and steroidogenic acute regulatory protein (StAR) (BMP-6 only; P<0.05). This supports the hypothesis that BMP-2 and -6 act as luteinization inhibitors. In conclusion, these findings provide evidence for the presence of a complex signalling mechanism in the porcine ovary and suggest that both BMP-2 and -6 may act in a paracrine manner to control granulosa cell function in this large polyovulatory species. PMID- 15826773 TI - Nutritional secondary hyperparathyroidism in rabbits. AB - The present study was designed to document the effect of a low (0.6%) calcium high (1.2%) phosphorus (LCaHP) diet on the development of parathyroid gland hyperplasia in rabbits and to describe the dynamics of parathyroid function (PTH Ca2+ curves) in rabbits with nutritional secondary hyperparathyroidism (N2HPT). Parathyroid gland weight, parathyroid cell proliferation (measured as percentage of cells in S-phase), and parathyroid calcium (CaRmRNA) and Vitamin D (VDRmRNA) receptor expression were measured in normal rabbits and in rabbits with N2HPT. The PTH-Ca2+ curve was studied in normal rabbits (Group I) and in rabbits with N2HPT at two stages: 2-3 weeks (Group IIA) and 5-6 weeks (Group IIB) after being fed LCaHP diet. An increase in parathyroid gland weight and percentage of cells in S-phase was detected in the course of N2HPT. After receiving a LCaHP diet for 6 weeks rabbits had decreased levels of CaRmRNA but VDRmRNA remained unchanged. A progressive increase in the concentrations of plasma PTH (Group IIA=167+/-14 pg/ml and Group IIB=377+/-54 pg/ml, P<0.05 versus Group I=27+/-3 pg/ml) was detected in the rabbits fed a LCaHP diet. This was accompanied by an increase in maximal and minimal PTH, reductions in plasma Ca2+ and calcitriol and elevations in plasma phosphate and creatinine. In conclusion, feeding a LCaHPD results in a rapid induction of N2HPT in rabbits. After 6 weeks on the LCaHPD rabbits develop parathyroid hyperplasia characterized by increases in PTH secretion, glandular weight and proliferation and by a decrease in CaRmRNA. PMID- 15826774 TI - Effects of exogenous cortisol on the GH/IGF-I/IGFBP network in channel catfish. AB - Glucocorticoids are known to hinder somatic growth in a number of vertebrate species. In order to better understand the mechanisms through which they may act in channel catfish, we examined the effects of feeding cortisol on the growth hormone (GH)/insulin-like growth factor-I (IGF-I)/IGF-binding protein (IGFBP) network. Fish (30.6+/-3.0 g) were fed once daily for 4 weeks and treatments included: (1) High-cortisol (dietary cortisol provided at 400 mg/kg feed), (2) Low-cortisol (dietary cortisol provided at 200 mg/kg feed), and (3) Control (commercial catfish feed). Fish fed diets with cortisol weighed approximately 50% less than Controls. Feed intake was reduced by approximately 30% in both treatments of cortisol fed fish compared to Controls. A approximately 20-kDa IGFBP was observed in plasma from High- and Low-treated fish while it was not detected in Control fish plasma. High-cortisol treatment increased pituitary GH mRNA expression approximately 10-fold while liver IGF-I mRNA expression was not different between cortisol-treated fish and Controls. Cortisol treatments decreased plasma levels of IGF-I. These data indicate that feeding cortisol for 4 weeks reduces weight gain, feed intake, and plasma levels of IGF-I and induces a approximately 20-kDa IGFBP. One mechanism through which cortisol may impede growth of catfish is through an increase in a low molecular weight IGFBP which may lead to inhibitory effects on the action of IGF-I. PMID- 15826775 TI - Dopamine receptors in equine ovarian tissues. AB - Dopamine (DA) agonist and antagonist treatments can affect ovarian reproductive events in the mare. To support our theory that DA produces these effects by acting directly on the ovary, we analyzed equine ovarian tissues for the presence of dopamine receptor-1 (D1r) and dopamine receptor-2 (D2r) mRNA by reverse transcription polymerase chain reaction (RT-PCR) and D1r and D2r proteins by Western blot and immunohistochemistry (IHC). RT-PCR was performed on RNA isolated from ovarian cortex, medulla, granulosa/theca or corpus luteum (CL) tissues and from pituitary (D2r control) and renal artery (D1r control). D1r and D2r specific primers were designed from partial DNA sequences known for the horse (D2r) or conserved sequences from other species (D1r). Western blot analyses were conducted on CL, cortex and granulosa/theca samples and IHC was performed on CL tissues using D1r or D2r specific antibodies. The incidence of positive D2r mRNA was high in CL and ovarian cortex, low in granulosa/theca, and not detectable in ovarian medulla. Dopamine D1r mRNA incidence was high (50%) only in CL tissues. D1r and D2r antibody staining was positive for each tissue type analyzed by Western blot procedures. All CL tissues prepared by IHC showed positive staining for D1r and D2r proteins. Both DA receptor proteins appeared uniformly distributed throughout the CL tissue. These results indicate that equine ovarian tissues do possess D1r and D2r, and suggests that DA can act directly on ovarian tissues through its interaction with DA receptors. PMID- 15826776 TI - Differences in reproductive pattern between wild and domestic rams are not associated with inter-specific annual variations in plasma prolactin and melatonin concentrations. AB - This study compares morphological changes, performed with real-time transrectal ultrasonography, of testis and accessory glands and seasonal endocrine changes in a wild (Mouflon) and a domesticated (Spanish Merino) breed of sheep. In Mouflons, the maximum plasma testosterone concentrations, testicular diameter and vesicular gland size occurred synchronously during autumn (P<0.001). In Merino rams the highest circulating levels of testosterone (P<0.05) and maximum testicular diameter (P<0.001) occurred during summer, with no seasonal variations in vesicular and bulbourethral glands. The seasonal changes in plasma concentrations of prolactin were not correlated with annual variations in testicular and glandular size in neither wild nor domestic species. No differences were observed between both species for the seasonal pattern of prolactin secretion and mean amplitude of melatonin. Wide differences in reproductive patterns between wild and domestic types of rams do not appear to be attributed to seasonal changes in prolactin and melatonin secretion. PMID- 15826777 TI - Effect of dietary supplementation of chitosan and galacto-mannan-oligosaccharide on serum parameters and the insulin-like growth factor-I mRNA expression in early weaned piglets. AB - The study was to determine effects of dietary supplementation of chitosan (COS) and galacto-mannan-oligosaccharides (GMOS) on some serum biochemical indices, serum growth hormone (GH) and insulin-like growth factor-I (IGF-I) levels, and hepatic and long gissimus muscle IGF-I mRNA expression in early-weaned piglets. Twenty six Duroc x Landrace x Yorkshire piglets at the age of 15 days were used. The piglets had access to creep feed during the suckling. Six piglets were sacrificed for sampling at the beginning of the study. The other 20 piglets were individually housed in metabolic cages and randomly allotted to four corn and soybean meal-based diets including the control group, the antibiotic group with 110 mg lincomycin/kg diet, the COS group containing 0.025% COS, and the GMOS group with 0.20% GMOS, respectively, in a 2-week feeding experiment. Blood urea nitrogen (BUN) level was reduced whereas serum total protein concentration was increased (P<0.05) in responses to the COS and GMOS supplementation. Dietary supplementation of COS and GMOS also increased (P<0.05) the serum GH and IGF-I levels along with enhanced hepatic and the muscle IGF-I mRNA abundance. Dietary supplementation of oligosaccharides such as COS and GMOS may improve growth and feed conversion efficiency by increasing plasma GH and IGF-I levels, in the early weaned piglets. PMID- 15826778 TI - Differential estradiol effects on estrogen and progesterone receptors expression in the oviduct and cervix of immature ewes. AB - In order to study the effect of estradiol-17beta (E2) on estrogen (E) and progesterone (P) receptors expression in oviduct and cervix of lambs, their respective transcripts (ERalpha mRNA and PR mRNA) were determined by solution hybridization and the receptor proteins (ER and PR) by binding assays after E2 treatments. Lambs (n=4 in each group) were not treated or treated with one, two or three i.m. injections of E2 (1 microg/kg) at 24 h of interval. Tissues were obtained 12 or 24 h after the last E2 injection. Estradiol treatments increased ERalpha mRNA and PR mRNA concentrations in an organ-dependent manner: transitory in the oviduct while maintained in the cervix. The E2 effect on the oviductal and cervical ER and PR concentrations were biphasic, with an initial reduction of receptors content that was followed by restoration. The ER restoration in oviduct was earlier than in the cervix. In summary, this study shows that E2 treatments may exert an inductive effect in ERalpha mRNA and PR mRNA levels and a biphasic effect in ER and PR concentrations in oviduct and cervix of immature ewe. These E2 effects varied in timing and strength depending on the organ of the reproductive tract. PMID- 15826779 TI - Stimulating effects of androgen on proliferation of cultured ovarian germ cells through androgenic and estrogenic actions in embryonic chickens. AB - The effect of androgen on germ cell proliferation was evaluated by a chicken ovarian germ-somatic cell co-culture model and the mechanisms were explored. Ovarian cells were dispersed from 18-day-old embryos, cultured in serum-free McCoy's 5A medium and challenged with testosterone (T) alone or in combinations with androgen receptor antagonist Flutamide, estrogen receptor antagonist Tamoxifen or aromatase inhibitor Letrozole for 48 h. Germ cells were identified by c-kit immunocytochemistry. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). The labeling index (LI) was determined for germ cells. Results showed that T (10(-7) to 10(-6)M) significantly increased the number of germ cells (P<0.05) and this stimulating effect was inhibited by Flutamide (10 1000 ng/ml), Tamoxifen (10-1000 ng/ml) or Letrozole (10(-9) to 10(-7)M) in a dose dependent manner. Furthermore, PCNA-LI of germ cells displayed similar changes with the numbers of germ cells. These results indicated that T-stimulated proliferation of cultured ovarian germ cells through both, androgenic and estrogenic actions in embryonic chickens. PMID- 15826780 TI - [After the passage to the year 2000, the passage of Afar at the year 2005]. PMID- 15826781 TI - [Surgical patient with severe sepsis: activated protein C and Surviving Sepsis Campaign]. PMID- 15826782 TI - [Selective decontamination of digestive tract: the light stays at orange]. PMID- 15826783 TI - [Activated protein C treatment: experience about 23 patients in the operative period]. AB - OBJECTIVE: To evaluate the use of activated C protein (ACP) in a Surgical Intensive Care Unit. STUDY DESIGN: A prospective observational study. PATIENTS AND METHODS: All patients receiving ACP during 20 months in the operative period. RESULTS: Twenty-three patients were treated by ACP. The origin of sepsis was peritonitis (n = 14), infected pancreatitis (n = 3), mediastinitis (n = 2), one urologic sepsis, one facial cellulitis, one catheter related infection, and one postoperative pneumonia. In two cases, the peritonitis was associated with a pleuretic infection, and in two other cases with parietal cellulites. Mean age was 69+/-13 years. Severities evaluated by SAPS II, LODS were 59+/-13 and 7+/-3, respectively. Mean number organ dysfunction was 3.3+/-1.0. Septic shock was present in 91% with concomitant use of catecholamines for a mean period of 87+/ 64 hours. Bacteraemia was present in 43% of the patients. A treatment with hydrocortisone was associated in 52% of the patients. The ICU and hospital lengths of stay were 15+/-16 days, and 34+/-38 days, respectively. Mortality at day 28 was 35%. Two significant bleeding were observed, one requiring red blood cell transfusion and the other one a surgical control of the bleeding associated with red blood cell transfusion. CONCLUSION: With global management of severe sepsis, including the use of activated C Protein, this prospective observational study showed a 30% reduction of the predicted mortality by SAPS II scoring without significant increase of bleeding episodes in a surgical context. PMID- 15826784 TI - [Low molecular weight heparin in patients undergoing vascular surgery]. AB - OBJECTIVES: Despite the fact that there are no approved indications by the French regulatory agency and despite the absence of recommendations, low molecular weight heparins (LMWH) are frequently used in vascular surgery, an area at both high risk of perioperative arterial and venous thrombosis as well as of bleeding. The aim of this study was to investigate medical practice in vascular surgery, using a survey of prescribers of antithrombotic agents. STUDY DESIGN: Survey of physicians from different specialities involved in patient care before, during, and after vascular surgery. PATIENTS AND METHODS: Between March and June 2003, 301 physicians filled a questionnaire providing information about their antithrombotic regimens depending on the type of vascular surgery. These physicians are involved in an important part of vascular surgery activity in France. RESULTS: The survey confirmed the use of LMWH in near 80% of patients scheduled for vascular surgery. The prescribed LMWH is frequently associated with antiplatelets agents. During patient's hospitalization, LMWH prescription is guided by numerous factors, making the prescription protocol rather complex. Major risk factors favoring continued prophylaxis after patient's hospital discharge include surgery with a high risk of thromboembolic events and limited ambulation. Analysis of the data also showed that the surgeon and the referring physician are both implicated in the majority of the postoperative care following patient's discharge, and therefore, in prescribing the antithrombotic drug. CONCLUSION: This survey gives information on practical use of LMWH in the vascular surgery setting. This information could lead to prospective studies of LMWH in situations other than venous thromboembolic disease. PMID- 15826785 TI - [Endovascular treatment of isthmic aortic rupture: use of second generation stent grafts]. AB - OBJECTIVE: Prospective analysis of endovascular management of traumatic isthmic rupture with second generation stent grafts. STUDY DESIGN: Prospective analysis and follow-up. PATIENTS: Ten consecutive multiple injured patients presenting an acute isthmic traumatic rupture who underwent an endovascular repair with second generation stent grafts. METHODS AND RESULTS: The aortic injury was diagnosed by spiral computed tomography scan. The appropriate time to repair was decided according to multidisciplinary decision after analysis of associated injuries status and mediastinal lesions evolution. Endovascular repair was successfully completed in all patients under general anaesthesia without requirement of haemodynamic manipulations. Despite a prolonged length of stay related to associated injuries, all patients were discharged from hospital without migration of devices or complication related to the endovascular procedure. After a 20 months follow-up (range 6 - 38 months), all patients were alive with a satisfactory CT scan analysis. CONCLUSION: The immediate availability of the second generation of stents-grafts allowed the endovascular treatment of isthmic rupture without haemodynamic manipulations or massive heparinization. The analysis of this selected series reinforces the interest of this non-invasive technique for anaesthetists especially in polytraumatized patients. PMID- 15826786 TI - [Role of Musca domestica in the transmission of multiresistant bacteria in the centres of intensive care setting in sub-Saharan Africa]. AB - OBJECTIVE: The houseflies (Musca domestica -MD) are involved in the transmission of the diseases of the faecal danger in subsaharan Africa. What is their role in the transmission of multiresistant bacteria in an intensive care setting in Dakar? TYPE OF STUDY: Descriptive, forward-looking. MATERIALS AND METHODS: The study was conducted from May 1 to September 30, 2003. During this period a flytrap was put above every patient carrier of BMR (methicillin resistant Staphylococcus, extended spectrum beta-lactamases-enterobacteria, ticarcillin resistant Pseudomonas). Caught MD were carried to the microbiology laboratory and incubated in heart-brain broth at 37 degrees C for 18 hours. Then selective gelose platers were used for identification of bacteria. Comparison of antibiotic sensitivities of bacteria isolated from MD and from the patients was made. RESULTS: Out of 441 hospitalized patients, 26 were colonised or infected by BMR. Human pathogenic microorganisms were obtained from 99 out of 120 flies. Seventeen of those flies were carried of BMR. Six among these 17 flies had BMR with sensitivity profile and phenotype of resistance identical to those of the patients under the trap with birdlime. More 3 MD carried BMR BLSE not found on our patients. CONCLUSION: This study shows that the MD can carry BMR. Their participation in the crossed transmission, between patients, of multiresistant bacteria in intensive care setting, must be discussed. The eradication of the MD in our Africa subsaharan services must be looked for. PMID- 15826787 TI - [Selective digestive decontamination: the light as changed from red to green]. AB - OBJECTIVES: To study the efficacy of selective digestive decontamination (SDD) for the prevention of nosocomial infections, particularly pneumonia, as well as its impact on the emergence of multiresistant bacteria. DATA SOURCES: Data collected from the Pubmed: original articles, review articles and editorial published on SDD. The keywords were: selective digestive decontamination, pneumonia, intensive care unit, infection. DATA SELECTION: Ten randomized clinical trials performed since 1995 in mechanically ventilated adult patients hospitalized in intensive care unit. RESULTS: The rationale for the use of SDD consists on the parenteral administration of a short course of antibiotic associated with the topical use of non-absorbable antibiotics directed against Gram negative bacteria. Five randomized studies described a reduction in the incidence of pneumonia associated with SDD. Only one study has showed a decrease in mortality rate. The other five studies, which present some methodological limitations, concluded the lack of efficacy of SDD. Regarding the emergence of multiresistant bacteria, the literature underlines the role of environment. The use of SDD seems to trigger the resistance in endemic areas, while these are softened in the units with a good control of their ecology. CONCLUSION: The data from the literature provide arguments to use SDD in targeted patient populations like multiple traumas in intensive care units, which have a low rate of multiresistant bacteria. PMID- 15826788 TI - The early steps of chloroform anaesthesia in Turkey during the Ottoman Empire in the 19th century. AB - OBJECTIVE: The aim of this study was to research the pioneering steps for the employment of chloroform in Turkey in comparison with the developments in the West i.e. in the United States and in Europe. The development of anaesthesiology in the West started in the first half of the 19th century. As an anaesthetic substance, ether was first employed in a medical operation by R. Liston in December 1846. But taking into consideration of its bronchially irritant effect, British gynaecologist Dr. J.Y. Simpson preferred to utilize chloroform in obstetrical operations in 1847. The paper aims at shedding light on the earlier steps for modern anaesthesiology in Turkey in that sense. METHODS: The survey used evaluation of archival documents, first hand-original sources such as the annual medical reports of the Medical School, books, official journals, and newspapers of the time, and also secondary sources concerned with the subject. RESULTS: In view of the findings of the survey, chloroform, as an anaesthetic material, began to be administered surgically in Turkey much earlier than it was already known. It was experienced and used in operations at the surgical clinic of the Imperial School of Medicine at the Capital city, Istanbul in 1848. The Crimean War (1853-1855) induced to the prevalent surgical use of chloroform in Istanbul on the soldiers back from the front. In other words, it was evidenced that surgeons started to make use of this anaesthetic substance in the Ottoman Empire, shortly after it was put into medical practice in Europe. CONCLUSION: This study deals with that phenomenal progress of chloroform anaesthesia in the medical history in Turkey during the second half of the 19th century. PMID- 15826789 TI - [Anaesthetic-induced myocardial preconditioning: fundamental basis and clinical implications]. AB - OBJECTIVE: Volatile halogenated anaesthetics offer a myocardial protection when they are administrated before a myocardial ischaemia. Cellular mechanisms involved in anaesthetic preconditioning are now better understood. The objectives of this review are to understand the anaesthetic-induced preconditioning underlying mechanisms and to know the clinical implications. DATA SOURCES: References were obtained from PubMed data bank (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi) using the following keywords: volatile anaesthetic, isoflurane, halothane, sevoflurane, desflurane, preconditioning, protection, myocardium. DATA SYNTHESIS: Ischaemic preconditioning (PC) is a myocardial endogenous protection against ischaemia. It has been described as one or several short ischaemia before a sustained ischemia. These short ischaemia trigger a protective signal against this longer ischaemia. An ischemic organ is able to precondition a remote organ. It is possible to replace the short ischaemia by a preadministration of halogenated volatile anaesthetic with the same protective effect, this is called anaesthetic PC (APC). APC and ischaemic PC share similar underlying biochemical mechanisms including protein kinase C, tyrosine kinase activation and mitochondrial and sarcolemnal K(ATP) channels opening. All halogenated anaesthetics can produce an anaesthetic PC effect. Myocardial protection during reperfusion, after the long ischaemia, has been shown by successive short ischaemia or volatile anaesthetic administration, this is called postconditioning. Ischaemic PC has been described in humans in 1993. Clinical studies in human cardiac surgery have shown the possibility of anaesthetic PC with volatile anaesthetics. These studies have shown a decrease of postoperative troponin in patient receiving halogenated anaesthetics. PMID- 15826790 TI - [Anaesthesia and prostate surgery]. AB - OBJECTIVES: To review the current data about anaesthetic management in prostate surgery with special regards on analysis and prevention of specific risks, appropriate anaesthetic procedure keeping with surgery and patient, recognition and treatment of adverse events. DATA SOURCES AND EXTRACTION: The Pubmed database was searched for articles (1990-2004) combined with references analysis of major articles on the field. DATA SYNTHESIS: It is strongly recommended to settle germfree urine in the preoperative period. The thromboembolic risk of radical retropubic prostatectomy for cancer parallels lower abdomen oncologic surgery and is prolonged. Preoperative evaluation of cardiovascular, respiratory, neurological and metabolic comorbidity is a source of prognostic information and an essential tool in the management of elderly patients with prostate disease. Extreme patient positioning applied in prostate surgery induces haemodynamic and respiratory changes and are associated with severe muscular and nervous injuries. The laparoscopic access for radical prostatectomy is a growing alternative to the open surgical procedure. Acute normovolaemic haemodilution is a consistent and cost-effective blood conservation strategy in reducing allogenic blood transfusion for radical retropubic prostatectomy. Whether open transvesical or transurethral prostatectomy for treatment of benign hypertrophy depends on the size of the gland: transurethral resection is safe up to 80 g. Intrathecal anaesthesia with a T9 cephalad spread of sensory block, produces adequate conditions for transurethral prostatectomy and allows a rapid diagnosis of irrigating fluid absorption syndrome. In spite of recommended preoperative antibiotic prophylaxis, bacteriemias are frequent during transurethral prostate resection. PMID- 15826791 TI - [Necrotizing bacterial skin and software tissue infection with muscular involvement revealing of cancer of the colon]. AB - We report the case of a 38-year-old woman with a necrotizing bacterial skin and soft tissue infection with muscular involvement. The clinical picture was similar to a gaseous gangrene of the right lower limb with a septic shock and multiple organ failure, without predisposing factor such as trauma, and necessitating a hip amputation. The primary site of the disease was a perforated colic adenocarcinoma with peritoneal and retroperitoneal infection. The association of necrotizing skin and soft tissue infection with muscular involvement due to Clostridium septicum to a neoplasma is classical and in front of such an infection a neoplasma should be researched. PMID- 15826792 TI - [Delayed and fortuitous diagnosis of right diaphragmatic rupture during preoperative evaluation]. AB - Although diaphragmatic rupture occurs after violent blunt trauma, its diagnosis is often delayed. The intra-thoracic displacement of abdominal organs through diaphragmatic rupture may lead to early or delayed complications because of compression of heart and lungs or strangulation of the abdominal viscera. A 49 year-old woman was scheduled to undergo elective lumbar disc surgery in prone position. The preoperative chest radiograph revealed an abnormally elevated right hemi diaphragm with loops of colon filling the right costophrenic angle. The CT scan confirmed right chronic diaphragmatic rupture with colon and liver herniations. Fifteen months previously, this woman had been involved in a traffic accident, with blunt right trauma resulting in right pelvic fractures. Perianaesthetic course increase the pre-existing mechanical risk of complications of chronic diaphragmatic rupture. Primary repair of right chronic diaphragmatic rupture through thoracotomy must be recommended. When another surgery must be done in emergency, the feasibility of regional anaesthesia should be considered. PMID- 15826793 TI - [Intraspinal air, a rare complication of blunt chest trauma]. AB - Pneumorachis, defined as the presence of air within the spinal canal, has rarely been described, and is exceptionally due to thoracic trauma. We report the case of a 37-year-old patient who sustained a motor vehicle accident. The chest CT scan showed a bilateral-hemothorax, a small right pneumothorax, rib fractures, and a fracture of the fourth thoracic vertebra associated with air in the spinal canal extending from T6 to L2. The fracture of T4 associated with the pleural lesion is probably responsible for the pneumorachis. Possible pathogenic mechanisms of this rare blunt chest complication are discussed. PMID- 15826794 TI - [Prolonged succinylcholine-induced apnoea associated with cholinesterase silent plasma variant]. AB - Mivacurium and succinylcholine are short-acting neuromuscular blocking drugs. The duration of their action depends on their rapid hydrolysis by plasma cholinesterase or butyrylcholinesterase. More than thirty genetic variants of this enzyme have been described but the atypical variant is the most frequently involved in prolonged apnoea following the administration of these two myorelaxants. We present one case of patient who presented a five hours long apnoea associated with null cholinesterase activity and detection of a butyrylcholinesterase silent variant (Sil-1, 1 for 100,000 persons in population) after sequencing. PMID- 15826795 TI - [Obstetrical anaesthesia and porphyrias]. AB - Acute hepatic porphyrias are genetic diseases, characterized by acute neurological symptoms, sometimes fatal, triggered by different factors, in particular by many anaesthetic drugs, and also by pregnancy. We report here the experience of three porphyric patients'deliveries, allowing us to consider a proposition of management in this context. After discussion between anaesthesiologist, obstetrician and porphyria specialist, two types of management of such patients can be foresee. Asymptomatic patients, or in long remission, can benefit from locoregional anesthesia techniques with bupivacaine for both labour analgesia and Caesarean section. Spinal anaesthesia is then the technique of choice, allowing using smaller quantity of local anaesthetic than epidural anaesthesia. For symptomatic patients, or in crisis, we have rather choose intravenous narcotics for labour analgesia, and general anaesthesia for Caesarean section. The hypnotic agent of choice for both induction and maintenance of such anaesthesia is then propofol. PMID- 15826796 TI - [Uncommon aetiology of peroneal compartment syndrome]. AB - Compartment syndromes are well recognized following major trauma. However, although uncommon, they may occur following athletic activity. We report a case of acute exertional peroneal compartmental syndrome in a 32-year-old that developed following horse riding. Because of the ignorance of pathology, a regional analgesia was carried out resulting in delayed diagnosis. Postoperative electromyography showed the absence of a fibula nerve compound action potential. At one-year follow-up visit following decompressive fasciotomy, muscular strength of the muscles of the anterior tibial compartment almost returned to normal. The presentation of this case of compartmental syndrome following horse riding allows to discuss the place of the regional anaesthesia. Because this anaesthesia technique can delay the diagnosis and the surgical treatment, it should not be used in first intention in the treatment of severe pain associated with compartmental syndrome. PMID- 15826797 TI - [Use of recombinant human activated protein C in severe sepsis in patient with a pericardial hemorrhagic collection after a recent cardiac surgery]. PMID- 15826798 TI - [Cardiac tamponade after internal jugular catheter malposition]. PMID- 15826799 TI - Crack cocaine as a cause of acute postoperative pulmonary edema in a pregnant drug addict. PMID- 15826800 TI - [Recommendations for management of severe sepsis and septic shock. Surviving sepsis campaign]. PMID- 15826801 TI - [Management of disseminated intravascular coagulation. Current practices]. PMID- 15826802 TI - [The preanesthetic visit "in the hours before the moment of the intervention": considerations]. PMID- 15826803 TI - Increased functional expression of P-glycoprotein in Caco-2 TC7 cells exposed long-term to cadmium. AB - The objective of this study was to investigate whether P-glycoprotein (P-gp) functional expression in intestinal cells is modified after long-term exposure to the food contaminant cadmium (Cd). The Caco-2 cell line, clone TC7, was first validated as a cellular model for long-term exposure to cadmium. Cytotoxicity tests after acute exposure of 24 h showed a significant concentration-dependent decrease in cellular viability at cadmium levels higher than 10 microM and led us to select the cadmium ranges for long-term exposure: 1, 5, and 10 microM. Intestinal cells were exposed to these cadmium concentrations for four consecutive weeks without inducing DNA condensation or fragmentation. In the second part of this work, we studied the functional expression of the drug efflux pump multidrug resistance P-glycoprotein after long-term exposure to cadmium by immunoblotting with the monoclonal antibody F4 and measurement of calcein-AM+/ the P-gp inhibitor verapamil. Western blot analysis with the F4 antibody detected a single band of 170 to 180 kDa which is the size previously reported for P-gp. Calcein-AM assay showed that four weeks exposure of intestinal cells to 1, 5, and 10 microM Cd increased P-gp functional expression in proportion to the Cd concentration. PMID- 15826804 TI - Estrogenic activity of 37 components of commercial sunscreen lotions evaluated by in vitro assays. AB - Thirty-seven chemical components of commercial sunscreen lotions were evaluated for estrogen agonistic and/or antagonistic activity using two in vitro assays, (1) an ELISA-based estrogen receptor competitive binding assay (ER-ELISA) and (2) a modified yeast two-hybrid estrogen assay, with and without addition of a rat liver preparation, S9 mix. Eleven compounds, most of which were benzophenone derivatives and parabens, showed binding affinity to ER by ER-ELISA without S9 mix. Although the activities of almost all of the compounds were attenuated by addition of S9 mix, 4-octylphenylsalicylate and 2,2'-dihydroxy-4,4' dimethoxybenzophenone acquired estrogenic activity, suggesting metabolic activation of these compounds. Two benzophenones showed agonistic activity in the yeast two-hybrid assay without S9 mix. The activity of one of these was reduced by S9 treatment and a further two benzophenones was activated. Eight parabens were active in this assay without S9 exposure, but their activities were eliminated by S9 treatment. Benzophenones with para-phenolic hydroxyl groups and parabens with branched and/or longer linear chains were generally more potent in both bioassays. In addition, weak antagonistic activity of 4-t-butylphenyl salicylate, 2-ethylhexyl 4-dimethylaminobenzoate and (+/-)-alpha tocopherolacetate was observed with S9 treatment. In vivo testing of the compounds reported here to have estrogen agonistic and antagonistic activities is required to confirm their effects at an organismal level. PMID- 15826805 TI - Effects of 6-hydroxydopamine on mitochondrial function and glutathione status in SH-SY5Y human neuroblastoma cells. AB - SH-SY5Y human neuroblastoma cells were incubated with 6-hydroxydopamine (6-OHDA) for 4 and 24 h to examine the mechanism of cell death and to determine the time dependent effects of 6-OHDA on cellular glutathione status. After 4 h, 6-OHDA significantly depleted cellular ATP and GSH concentrations with only slight increases in cell death. GSH:GSSG ratios and mitochondrial membrane potential (Deltapsim) were significantly decreased during 4 h incubations with 6-OHDA. High concentrations of 6-OHDA (100 microM) induced oxidative stress and mitochondrial dysfunction in SH-SY5Y cells within 4 h leading to cell death. In 24 h incubations, 25 and 50 microM 6-OHDA significantly decreased ATP concentrations; however, significant increases in cell death were only observed with 50 microM 6 OHDA. 6-OHDA induced a concentration-dependent increase in GSH and total glutathione concentrations after 24 h. After exposure to 50 microM 6-OHDA, GSH concentrations were increased up to 12-fold after 24 h with no change in the GSH:GSSG ratio. Gene analysis suggests that the increase in GSH concentration was due to increased expression of the GSH synthesis genes glutamate cysteine ligase modifier and catalytic subunits. Our results suggest that 6-OHDA induces oxidative stress in SH-SY5Y cells resulting in an adaptive increase in cellular GSH concentrations. PMID- 15826806 TI - Cadmium chloride-induced DNA and lysosomal damage in a hepatoma cell line. AB - Cadmium is a toxic metal and no uniform mechanism of toxicity has so far been proposed. The aim of this study was to investigate the biochemical effects of cadmium chloride in a rat hepatoma cell line (HTC cells) and the cellular events mediating DNA damage. HTC cells were exposed to various concentrations of cadmium chloride for 5 and 8 h and lysosomal damage was assessed with the neutral red assay (NR) and fluorescence microscopy. Mitochondrial integrity was assessed from ATP levels and DNA damage determined with the single cell gel electrophoresis/comet assay. The formation of reactive oxygen species (ROS) was also determined under the same experimental conditions with the dichlorofluorescein assay. Cytotoxicity was assessed with the LDH leakage assay and the levels of glutathione were measured and correlated with the other effects. The results indicate that lysosomal damage occurs at a lower concentration of cadmium chloride (20 microM) than DNA damage (500 microM) in HTC cells. The latter effect was accompanied by an increase of reactive oxygen species without any significant LDH leakage whereas lysosomal damage was significant as determined by the neutral red assay and confirmed with fluorescence microscopy. The effect of CdCl2 on mitochondria and glutathione levels were observed at concentrations or incubation times higher than the ones required to induce lysosomal damage. The data suggest that DNA damage may be due to the formation of reactive oxygen species. It is possible that cadmium induced lysosomal damage is an earlier event than DNA damage and can mediate other cellular events that lead to cell death. PMID- 15826807 TI - Cytotoxic effects of 110 reference compounds on HepG2 cells and for 60 compounds on HeLa, ECC-1 and CHO cells. II mechanistic assays on NAD(P)H, ATP and DNA contents. AB - In this study the focus is on the comparison of fluorometric assays, using Alamar Blue (AB) and Hoechst 33342 coloration, and luminometric assays, using Cyto-Lite and ATP-Lite, for toxicity measurements. With AB, ATP-Lite and Cyto-Lite the energy status of the cell is measured and with Hoechst 33342 the amount of DNA. These assays were carried out with different dosages of several toxic compounds with the following permanent cell lines: human liver (Hep G2), human endometrium (ECC-1), human cervix (HeLa) and Chinese hamster ovary (CHO) cells. In these assays toxicity of 110 compounds was assessed in Hep G2 cells. With 60 of those, toxicity was assessed in Hela, ECC-1 and CHO cells. These compounds were non narcotic antitussives, nasal decongestants, narcotic analgesics, hypnotics, vasodilators, specific cellular energy blockers, cellular proliferation inhibitors, ion channel blockers, estrogens, antiestrogens, androgens, progestagens and others. The outcome of this study is that all four cell lines were responsive to the same set of 60 drugs with a comparable indication of toxicity. Hep G2 cells appear slightly more sensitive, as compared to the other three cell lines. Evaluation up to dosages of 3.2 x 10(-4) or even 3.2 x 10(-3)M for some of the compounds for these four assays in Hep G2 cells demonstrated toxicity for 45 of the 60 (75%) reference compounds with known toxicity in these assays. With a new set of 50 compounds, among which there were estrogens, androgens, progestagens and antiestrogens, 18 (36%) were identified as toxic up to a concentration of 3.2 x 10(-5)M. In conclusion, many of the 60 tested reference compounds gave similar dose and toxicity effects on these permanent cell lines. Therefore, all these cell lines can be used for toxicity screening with AB, ATP-Lite, Cyto-Lite and Hoechst 33342. However, species specific cell lines may reveal species specific effects, as shown with digoxin. PMID- 15826808 TI - Cytotoxic effects of 100 reference compounds on Hep G2 and HeLa cells and of 60 compounds on ECC-1 and CHO cells. I mechanistic assays on ROS, glutathione depletion and calcein uptake. AB - In this study fluorometric assays have been used for medium throughput screening on toxicity. Dichlorofluorescein diacetate, monochlorobimane and calcein-AM were fluorophores for the measurement of the formation of reactive oxygen species (ROS), the quantification of glutathione and the membrane stability, respectively. These assays have been carried out in the presence or absence of toxic compounds and with four different cell lines, i.e. human liver (Hep G2), human endometrium (ECC-1), human cervix (HeLa) and Chinese hamster ovary cells (CHO). In these assays the toxic dose of 60 reference compounds was assessed for Hep G2, HeLa, ECC-1 and CHO cells and of 40 pharmaceutical compounds for Hep G2 (ROS, glutathione) or HeLa (calcein) cells, only. These compounds were narcotic analgesics, hypnotics, vasodilators, specific cellular energy blockers, cellular proliferation inhibitors, ion channel blockers, estrogens, antiestrogens, androgens, progestagens and others. The outcome of this study revealed that all four cell lines were responsive to the same set of drugs. Only for some drugs Hep G2 cells appear slightly more sensitive, as compared to the other three cell lines. In general the HeLa cell line was the most sensitive cell line for the calcein uptake, while the Hep G2 cell line shows slightly more sensitivity for dichorofluorescein and monochlorobimane assays than the other three cell lines. Further evaluation at higher toxic dosages with Hep G2 cells for ROS and glutathione depletion and HeLa cells for calcein uptake, demonstrated toxic effects for 56 of the 100 reference compounds in these assays, among which there were estrogens, androgens, progestagens and antiestrogens. In conclusion, almost all tested compounds gave similar dose and toxicity effects on the permanent cell lines used in this study. Only three compounds showed more tissue specific cell responses. This shows that in principle all four cell lines can be used for toxicity screening. PMID- 15826809 TI - Molecular modelling of human microsomal epoxide hydrolase (EH) by homology with a fungal (Aspergillus niger) EH crystal structure of 1.8 A resolution: structure activity relationships in epoxides inhibiting EH activity. AB - Homology modelling of the human microsomal epoxide hydrolase (EH) enzyme based on the fungal (Aspergillus niger) EH crystallographic template is reported. The active site lies in a well-defined, essentially hydrophobic, pocket within the overall enzyme structure. Two tyrosine residues, that are conserved in all known mammalian EH sequences, are able to form hydrogen bonds (one per tyrosine residue) with the epoxide oxygen atom on the known EH substrate, styrene oxide. There is also a small hydrophobic cleft, within the active site region, where the phenyl group of styrene oxide can bind, but this appears to be restricted such that the presence of bulky side-chains will render poor substrate status to the incoming epoxide molecule. Quantitative structure-activity relationship (QSAR) studies on a series of low molecular weight epoxides provide useful results which appear to be generally consistent with the human microsomal EH model, and thus may be used predictively for assessing the EH substrate and/or inhibitor status of untested compounds. PMID- 15826810 TI - Induction of apoptosis and inhibition of telomerase activity in human bone marrow and HL-60 p53 null cells treated with anti-cancer drugs. AB - Telomerase plays a key role in the maintenance of chromosomal stability in tumours, and the ability of anti-cancer agents to inhibit telomerase activity is under investigation. In this study, we evaluated the effect of etoposide and taxol, on the telomerase activity and telomere length in human leukaemia p53 null cells and human bone marrow cells, as well as apoptosis and cell cycle modulation. Results showed that after exposure to the drugs, HL-60 cells as well as the human progenitors underwent a block in G2 and subsequently apoptosis, whereas stromal cells from bone marrow did not undergo a block in G2 or enter apoptosis after etoposide exposure. Telomere length increased in stromal cells after treatment with both etoposide and taxol whereas in HL-60 cells only after etoposide treatment with. Bax, bcl-2 and bcl-x change their expression in stromal cells, whereas bcl-x was induced after drug treatment and bcl-2 down regulated in progenitor cells. Our data suggest that telomerase activity and apoptosis are correlated and they seem to be modulated by a common gene, bcl-2. PMID- 15826811 TI - Cytotoxicity, genotoxicity and antimutagenicity of hexane extracts of Agaricus blazei determined in vitro by the comet assay and CHO/HGPRT gene mutation assay. AB - Agaricus blazei Murrill ss. Heinem, known as the sun mushroom or himematsutake, is a basidiomycete native to Brazil, which is popular for its medicinal properties. The aim of this study was to test hexane extracts (one fraction and its four sub-fractions) of A. blazei for bioactivity in cultured mammalian cells (CHO-K1). The comet assay, the colony forming assay (CFA) and CHO/HGPRT gene mutation assay were used respectively to determine genotoxicity, cytotoxicity and antimutagenicity of these extracts at different concentrations. The cells were incubated in culture medium and treated for 3h according to the standard protocol for each assay. The DNA damage-inducing agent ethylmethane sulfonate (EMS) was utilized as the positive control and also in combination with extracts to test for a protective effect. Statistical analysis of the data was performed using analysis of variance (ANOVA) and Tukey's test. A relationship between cytotoxicity and genotoxicity could be established and two extracts EH6B and EH6D showed a protective tendency, while the others did not, with the primary extract EH6 causing the most substantial damage to genetic material. These findings warrant more in-depth studies of the active principles of this mushroom. PMID- 15826812 TI - Effect of the immunosuppressive regime of Edmonton protocol on the long-term in vitro insulin secretion from islets of two different species and age categories. AB - The success of the new immunosuppressive regime known as the Edmonton protocol in islet allotransplantation may suggest that it is also possible that this regime may prevent the rejection of xenografts. This protocol applies a combination of Tacrolimus, Sirolimus and Daclizumab at low doses. This combination may have some toxicity that affects the function and viability of the pancreatic islets. The choice of species or age category, whose islets can tolerate the toxicity of this immunosuppressive combination, may become important for the graft survival. It was the aim of this study to investigate the long-term effect of this regime on insulin secretion from pancreatic islets isolated from two species (rats and pigs) and from two age categories (day 7 postnatal [P7] and adult rat). Islets were cultured for three weeks in medium containing Tacrolimus in a concentration of 5 ng/ml, while the concentration of Sirolimus was 15 ng/ml. Daclizumab was added at the beginning of culture and once weekly in a final concentration of 10 ng/ml. In immunosuppressive-treated groups, Glucose was able to stimulate increases of insulin secretion over the basal value after 1 and 3 weeks in adult rat islets, and could not stimulate this secretion in P7 islets, while it stimulated the secretion only after 1 week, but not 3 weeks, in porcine islets. The immunosuppressive regimen caused significant reductions of glucose-stimulated insulin secretion magnitude in adult rat and porcine islets after 3 weeks, while it reduced both basal and stimulated secretions after 1 and 3 weeks in P7 rat islets. There was no difference in DNA contents between control and immunosuppressive-treated groups after 1 or 3 weeks in any of the islet preparations. DNA decreased considerably with the time in culture. The change in DNA content over 3 weeks was higher in the Edmonton group of adult porcine and P7 rat than in adult rat islets. Comparison of the responses of islets from different age categories and species leads to conclude that in vitro cultures of adult rat islets are more tolerant to this immunosuppressive combination toxicity than P7 islets, and there is variable responses of islets from different adult species to this toxicity. PMID- 15826813 TI - In vitro genotoxic evaluation of three alpha-asarone analogues. AB - alpha-Asarone has shown a significant capacity to reduce the level of lipids, including cholesterol. However, several toxic and genotoxic studies have determined that its use may pose a risk to human health. Therefore, a series of compounds structurally analogous to alpha-asarone were prepared in order to maintain the same pharmacological properties but with low toxicity. In this study we evaluated the potential of three alpha-asarone analogues to induce mutagenicity using the Ames test (strains TA98 and TA100 in the presence of metabolic activation), as well as the induction of sister chromatid exchanges (SCE) in cultured human lymphocytes. The tested compounds were: 1-(2,4,5 trimethoxyphenyl)propan-1-one (D1), 1-(2-chloro-4,5-dimethoxyphenyl)propan-1-one (D2), and 1-(4,5-dimethoxy-2-nitrophenyl)propan-1-ol (D3). The results in the first assay showed no mutagenic effect for the three tested analogues; in the TA100 strain, certain cytotoxicity did appear in the case of D2 and D3 only at high concentrations. In regard to the SCE assay, compounds D1 and D2 presented no statistical differences in comparison with the control culture values; however, the high dose of D3 (300 microg/ml) produced a significant increment in SCE (68% above the control value). With respect to the mitotic index and the cellular proliferation kinetics, we observed a reduction when compounds D2 and D3 were used at the higher concentrations. Our results encourage further preclinical studies of these compounds in both in vitro and in vivo models (particularly for analogues D1 and D2), to determine their toxicological profile and establish the possibility of using them in humans. PMID- 15826814 TI - Toxicity of fatty acids on murine and human melanoma cell lines. AB - High concentrations of certain fatty acids can cause cell death via apoptosis or necrosis. The aim of this study was to investigate the toxicity of saturated and unsaturated fatty acids on melanoma cell lines, which was evaluated by either loss of membrane integrity and/or DNA fragmentation using flow cytometric analysis. Evidence is presented that saturated and unsaturated fatty acids exert toxic effects on melanoma cells through loss of membrane integrity and/or DNA fragmentation. Arachidonic and linoleic acids were the most effective in decreasing the number of viable S91 murine melanoma cells, causing loss of membrane integrity and DNA fragmentation at 100 microM concentration already after 24 h in culture. In B16F10 murine melanoma cells, palmitic acid was the most toxic, inducing cell death by both apoptosis and necrosis. The human melanoma cell lines were more resistant to the toxic effect of fatty acids. In SK Mel 23 cells, indications of cytotoxicity were detected only after 48 h treatment with arachidonic, linoleic, palmitic and palmitoleic acids at 200 microM concentration. Linoleic acid was the most toxic for this cell line. In SK-Mel 28 human cells, only palmitic acid caused a significant decrease of the number of viable cells, inducing DNA fragmentation after 24 and 48 h treatments at 200 microM concentration. PMID- 15826815 TI - VEGF, IL-18 and NO production by neutrophils and their serum levels in patients with oral cavity cancer. AB - It is known that the relationship between pro-angiogenic and anti-angiogenic factors is responsible for the presence and intensity of neoangiogenesis. The angiogenic factors are produced by tumour cells and/or by tumour-infiltrating inflammatory cells such as macrophages or polymorphonuclear leukocytes (PMN). In the present study we compared VEGF secretion with IL-18 and NO release by PMN derived from oral cavity cancer patients. Knowledge of the relationship between mediators above could help in better understanding the role of PMN in angiogenesis in this patient group. The results from culture supernatants of PMN were confronted with the serum levels of parameters examined. We found an interesting relationship between VEGF and IL-18 concentrations in the culture supernatants of PMN derived from patients with oral cavity cancer. High production of VEGF was associated with low production of IL-18 by PMN derived from patients before treatment. During examinations after treatment we found lower concentrations of VEGF and higher concentrations of IL-18 than those in the study before treatment. In contrast to VEGF and IL-18, the NO production by PMN of cancer patients, before and after treatment, was unchanged. We also demonstrated markedly elevated serum levels of VEGF as well as IL-18 according to the progression of the disease. Results obtained indicate that relations between VEGF and IL-18 released by PMN may promote neoangiogenesis and may be important for benign tumour cells to acquire metastatic phenotype in the early stage of oral cavity cancer. Furthermore, our results suggest that the concentrations of VEGF and IL-18 in the serum are sensitive tumour markers in this patient group before and after treatment. PMID- 15826816 TI - Treatment of mice with IL-12 DNA constructs leads to augmented NK activity in lungs but low IFN-gamma release -- implications for Bordetella pertussis infections following aerosol challenge. AB - Interleukin-12 protein has been widely used experimentally in therapeutic and adjuvant settings in the treatment of different diseases including intra-cellular bacterial infections. The in vivo clearance of Bordetella pertussis infections in naive mice and in animals vaccinated with whole cell vaccine is considered to be a Th-1 dependent mechanism. Furthermore, the addition of IL-12 protein to an acellular pertussis vaccine increases the efficacy of this vaccine. Whilst the use of IL-12 protein is often beneficial, a number of problems there are associated with this cytokine including toxicities and down regulation of normal immune functions. The use of DNA constructs encoding this cytokine may be a way of achieving maximum therapeutic benefit with minimum toxicity. The aims of this study were to optimise the effects of two IL-12 DNA constructs, especially with respect to augmenting pulmonary immune responsiveness and to compare the effect of IL-12 DNA and IL-12 protein on bacterial colonisation of lungs following aerosol challenge with B. pertussis. We found that IL-12 DNA constructs augmented the activity of pulmonary NK cells but had little effect on the course of B. pertussis infections in mice. In contrast to IL-12 protein, the DNA constructs had no immunosuppressive effects on splenic lymphocyte mitogen responses. PMID- 15826817 TI - The impact of burn injury and ethanol on the cytokine network of the mouse hypothalamus: reproductive implications. AB - Nearly 50% of the patients admitted to hospitals for burn injuries have detectable levels of alcohol (EtOH) in their circulation. In fact, EtOH is often a causal factor in their injury. It is well known that EtOH as well as burn injury disrupt function of the hypothalamic-pituitary-gonadal (HPG) axis. The cellular mechanisms by which EtOH and/or burn impacts on the HPG are not entirely understood. In the studies reported here, we tested the hypothesis that these injuries mediated their effects by local hypothalamic inflammation. Young adult male mice were subjected to either a 15% total body surface area, full thickness scald, to EtOH, or to both and compared to appropriate controls. They were sacrificed 48 h later. EtOH and burn, as well as the combined injury, consistently and impressively reduced serum testosterone, while increasing hypothalamic concentrations of all three of the pro-inflammatory cytokines, TNFalpha, IL-1beta, and IL-6. In general, the increases induced by burn were greater than those caused by EtOH and the effect of the combined insult was not additive. Hypothalamic concentrations of LHRH were also increased. The data are consistent with the idea that EtOH and/or burn, as models of critical illness, medicate their hypothalamic suppressive effects via increase in pro-inflammatory cytokines. PMID- 15826818 TI - Interleukin 4 induces the expression of inducible nitric oxide synthase in eosinophils. AB - We investigated the effects of the Th2-like cytokines IL-4 and IL-13 and of IL-10 on the induction of iNOS and NO production in rat eosinophils. Addition of mIL-4 to the eosinophil culture increased iNOS activity and nitrite production but did not improve the stimulatory effect of IFN-gamma and LPS. In contrast to eosinophils, addition of mIL-4 to macrophage cultures inhibited the iNOS expression and nitrite production induced by IFN-gamma plus LPS. Addition of mIL 13 to the eosinophil cultures did not significantly change iNOS activity and nitrite production in cells stimulated or not with IFN-gamma plus LPS. On the other hand, IL-13 inhibited iNOS activity in IFN-gamma plus LPS-stimulated macrophages. In the presence of IL-10, iNOS activity in non-stimulated eosinophil or macrophage cultures was not significantly altered, but the enzyme expression was inhibited in IFN-gamma plus LPS-stimulated eosinophils or macrophages. The production of nitrite by eosinophils stimulated by IFN-gamma plus LPS was inhibited by the presence of IL-10 in the medium. In conclusion, eosinophils might exhibit differential modulation of the L-arginine/iNOS pathway depending on the profile of Th2 cytokines produced during allergic diseases. IL-4 appears to be an important Th2 cytokine involved in the induction of the L-arginine/iNOS pathway in eosinophils. PMID- 15826819 TI - Neuroprotection with the CXCL8 inhibitor repertaxin in transient brain ischemia. AB - Infiltration of polymorphonuclear neutrophils (PMNs) is thought to play a role in ischemic brain damage. The present study investigated the effect of repertaxin, a new noncompetitive allosteric inhibitor for the receptors of the inflammatory chemokine CXC ligand 8 (CXCL8)/interleukin-8 (IL-8), on PMN infiltration and tissue injury in rats. Cerebral ischemia was induced by permanent or transient occlusion of the middle cerebral artery and myeloperoxidase activity, a marker of PMN infiltration, and infarct volume were evaluated 24 h later. Repertaxin (15 mg/kg) was administered systemically at the time of ischemia and every 2 h for four times. In permanent ischemia repertaxin reduced PMN infiltration by 40% in the brain cortex but did not limit tissue damage. In transient ischemia (90-min ischemia followed by reperfusion), repertaxin inhibited PMN infiltration by 54% and gave 44% protection from tissue damage. Repertaxin had anti-inflammatory and neuroprotective effects also when given at reperfusion and even at 2 h of reperfusion. The protective effect of repertaxin did not interfere with brain levels of the chemokine. Since the PMN infiltration and its inhibition by repertaxin were comparable in the two models we conclude that reperfusion induces PMN activation, and inhibition of CXCL8 by repertaxin might be of pharmacological interest in transient ischemia. PMID- 15826820 TI - Chronic inflammation and hemodialysis reduce immune competence of peripheral blood leukocytes in end-stage renal failure patients. AB - Immunoincompetence is a profound problem in end-stage renal failure patients undergoing hemodialysis, and chronic inflammation with altered serum levels of inflammation markers has been reported. Gene expression patterns have had little relevance for leukocyte research so far because of limitations in transcript levels and stability. Using a new stimulation system we induced the expression of immune-relevant transcripts in whole blood preparations ex vivo and stabilized transcript levels by preventing RNA degradation and uncontrolled gene induction. Using quantitative real-time PCR we could show that basal TGF-beta mRNA expression is about 2-fold decreased in end-stage renal failure patients, while expression of TNF-alpha becomes 2-fold increased, further doubling during hemodialysis. By short term stimulation with phytohemagglutinin (PHA) for 2 h we tested for the immune competence of peripheral blood leukocytes and demonstrated that hemodialysis decreases TNF-alpha-mediated immune responsiveness more than 3 fold. This study shows by induction and stabilization of immune-relevant transcripts that chronic inflammation and hemodialysis are crucial factors for disturbed immune competence of end-stage renal failure patients. PMID- 15826822 TI - Models for peripheral B cell development and homeostasis. AB - Immature B cells undergo key maturation and selection events after migrating to peripheral lymphoid organs. We will review recent advances in our understanding of the cell populations and molecular interactions underlying the differentiation of immature peripheral B cells into mature marginal zone (MZ) and follicular B cells, and discuss potential mechanisms by which numbers of MZ and follicular B cells are maintained. We will also discuss current controversies over the identity of precursor cells for MZ and follicular B cells, and propose a potentially unifying model for precursor-product relationships in peripheral B cell maturation. PMID- 15826823 TI - Regulation of mature T cell homeostasis. AB - The overall size and the composition of the mature T cell pool are regulated by homeostatic mechanisms. Recent work has revealed that homeostatic signals are received from contact with two members of the common gamma chain family of cytokines, IL-7 and IL-15, and from self-MHC/peptide ligands. In essence, homeostasis of naive T cells is regulated by IL-7 and self-MHC/peptide ligands and homeostasis of memory CD8 cells is controlled by IL-7 and IL-15. All of these signals also appear to be important to a varying degree for homeostasis of memory CD4 cells, but the details are less well understood than for other cell type. PMID- 15826824 TI - Homeostatic niche specification among naive and activated B cells: a growing role for the BLyS family of receptors and ligands. AB - B lymphocyte homeostasis encompasses the establishment and maintenance of independently regulated niches, within which cells compete for viability promoting resources. The BLyS/BLyS receptor family controls the size and composition of these niches, by governing the selection and survival of most peripheral B cells. Moreover, different receptor-ligand sets from this family dominate the regulation of various B cell subsets. These observations suggest a model whereby the regulation of BLyS receptors by differentiative and stimulatory cues yield characteristic BLyS receptor signatures, thus specifying homeostatic niche and competitive advantage. PMID- 15826825 TI - Endogenous proliferation: burst-like CD4 T cell proliferation in lymphopenic settings. AB - Rapid and slow proliferation is observed when naive CD4 T cells are transferred into lymphopenic hosts. We have recently proposed that the rapid, burst-like proliferation, designated endogenous proliferation, is a peripheral mechanism by which memory T cells of diverse specificity are generated without exogenous antigenic stimulation. In this review, we discuss some of unique features of endogenous proliferation. We argue that it is regulated not by the absolute number of memory cells present but by the range of specificities of those cells. We discuss the physiologic significance of endogenous proliferation and outline goals for future studies. PMID- 15826826 TI - Homeostatic proliferation of B cells. AB - Naive B cells introduced into a lymphopenic host undergo antigen-independent proliferation which is inhibited in a cell dose dependent manner by feedback from mature B cells. Homeostatic proliferation is a generalized lymphocyte property with B cells sharing many of the inductive and regulatory characteristics established for naive and memory CD4+ and CD8+ T cells and NK cells. In this communication we discuss the cytokine requirements for B cell HP, extend the murine studies to human cells, and propose the hypothesis that B cell HP may provide an antigen-independent mechanism for maintaining B cell memory. PMID- 15826827 TI - The CD8 memory T cell subsystem: integration of homeostatic signaling during migration. AB - The ability of memory CD8 T cells to patrol non-lymphoid tissues represents an effective method whereby proficient immunosurveillance is achieved. From the analysis of memory CD8 T cell migration in vivo, it is clear that tissue-specific factors control trafficking and residence time within tissues. We propose that at least three pools of memory CD8 T cells exist based on migratory capabilities as dictated by their location in the body. Moreover, we hypothesize that the process of acquisition of homeostatic signals in specific tissues, such as the cytokines IL-7 and IL-15, regulates the mobility of memory T cells. PMID- 15826828 TI - T cell homeostasis: keeping useful T cells alive and live T cells useful. AB - It has become clear that the regulation of T cell numbers is under peripheral homeostatic control. However, the rules for homeostasis vary with the T cells' differentiation state and the overall T cell number in the animal. Furthermore, homeostatic pressures can cause unexpected changes in T cell differentiation and function which might promote or dampen T cell reactivity. In this review, we focus on the role of peptide/MHC and cytokine interactions in regulating the size and composition of the T cell pool. PMID- 15826829 TI - Homeostasis of T cell numbers: from thymus production to peripheral compartmentalization and the indexation of regulatory T cells. AB - A system under homeostatic control tends to maintain its structure and functions by establishing dynamic equilibriums controlled by multiple regulatory mechanisms. We have shown that this is the case for immune system. Several different mechanisms seem to participate in the homeostatic control of T cell numbers and population distribution. In other words, besides a quantitative dimension, there is also a qualitative dimension in T cell homeostasis. This is achieved through competition by driving the specialization of sub-populations of lymphocytes to occupy specific niches in the peripheral pool and by developing independent homeostatic mechanisms for each particular cell sub-set. Thus, the sizes of the naive and memory T cell compartments are governed by independent homeostatic mechanisms, which preserve the capacity to deal with any novel infection (conferred by the presence of naive T cells) whilst ensuring the efficacy of memory responses when dealing with recurring antigens. Peripheral T cell homeostasis also depends on the integrity of sub-population structure and the presence of regulatory CD4+ CD25+ T cells. The indexation of regulatory CD4+ CD25+ T cell numbers to the numbers of peripheral activated CD4+ T cells is another mechanism of homeostasis that has major advantages in the control of immune responses. It ensures continuous regulation of T cell numbers throughout immune responses, allowing for increases in cell numbers as long as the proportion of CD4+ CD25+ regulatory T cells is kept. PMID- 15826830 TI - Fusion oncogenes. PMID- 15826831 TI - ETV6: a versatile player in leukemogenesis. AB - Alterations of the ets family transcription factor ETV6 (TEL) and the RUNT domain transcription factor RUNX1 (AML1) play pivotal roles in the leukemogenesis of various types of leukemia. While only three fusion partners of RUNX1 namely ETO, ETV6 and MTG16 have been described so far, there is a plethora of ETV6 fusion partners with about 20 partners described so far. Apart from forming fusion genes there are other genetic alterations of ETV6 including deletions, point mutations and possible alterations at the promoter level that might contribute to the malignant phenotype. This review will focus on ETV6 and on the different mechanisms that are used by this gene to cause leukemia. PMID- 15826832 TI - The versatile mixed lineage leukaemia gene MLL and its many associations in leukaemogenesis. AB - The marked association of abnormalities of chromosome 11 long arm, band q23, with human leukaemia led to the identification of the 11q23 gene called MLL (or HTRX, HRX, TRX1, ALL-1). MLL can become fused with one of a remarkable panoply of genes from other chromosome locations in individual leukaemias, leading to either acute myeloid or lymphoid tumours (hence the name MLL for mixed lineage leukaemia). The unusual finding that a single protein could be involved in both myeloid and lymphoid malignancies and that the truncated protein could do so as a fusion with very disparate partners has prompted studies to define the molecular role of MLL fusions in leukaemogenesis and to the development of MLL-controlled mouse models of leukaemogenesis. These studies have defined MLL-fusion proteins as regulators of gene expression, controlling such elements as HOX genes, and have indicated a variety of mechanisms by which MLL-fusion proteins contribute to leukaemogenesis. PMID- 15826833 TI - Context matters: the hen or egg problem in Ewing's sarcoma. AB - Ewing's sarcoma and related tumors (ESFT) are characterized by rearrangements of EWS with ets family genes. While detection of these gene fusions greatly facilitated diagnosis, it has not provided any clues about the tissue of origin. Immunological and gene expression profiling studies favour a neuroectodermal histogenesis. These investigations did not appreciate the impact of EWS-ets proteins on the tumor phenotype. Introduction of EWS-ets into different cellular models resulted in diverse outcomes ranging from the induction of cell cycle arrest or apoptosis to transformation and tumorigenicity, and from blocked differentiation to trans-differentiation. Thus, the molecular signature of EWS ets proteins depends on the cell type. The hen or egg problem in ESFT, therefore, is whether ESFT reflect the phenotype of the tumor stem cell that is blocked in differentiation by the activity of the EWS-ets gene fusion or if the oncogene imposes an incomplete differentiation program on a pluripotent precursor cell. This article addresses the problem by considering the tissue distribution of FLI1 and ERG expression and by reviewing evidence for combinatorial control of EWS-ets activity. PMID- 15826834 TI - The EWS-WT1 gene fusion in desmoplastic small round cell tumor. AB - Desmoplastic small round cell tumor (DSRCT) is a poorly understood neoplasm with distinctive clinical, histologic and immunophenotypic features. It is associated with a novel, specific chromosomal abnormality, t(11;22)(p13;q12) that fuses EWS with WT1 leading to production of a chimeric protein with transcriptional regulatory activity. This chimeric transcription factor has unique DNA-binding properties and regulates expression of specific target genes. Several of these have been identified and their biological role characterized. The dysregulated expression of EWS-WT1 targets contribute to the malignant phenotype of DSRCT and provide valuable insight regarding the molecular mechanisms underlying the development and progression of this distinct translocation associated tumor. PMID- 15826835 TI - Understanding mesenchymal cancer: the liposarcoma-associated FUS-DDIT3 fusion gene as a model. AB - Chromosomal translocations entail the generation of gene fusions in mesenchymal tumors. Despite the successful identification of these specific and consistent genetic events, the nature of the intimate association between the gene fusion and the resulting phenotype still remains to be elucidated. Here these studies are reviewed, using FUS-DDIT3 as a model to illustrate how they have contributed to current understanding in unique and unexpected ways. FUS-DDIT3 is a chimeric oncogene generated by the most common chromosomal translocation t(12;16)(q13;p11) associated with liposarcomas. The application of transgenic methods to the study of this sarcoma-associated FUS-DDIT3 gene fusion has provided insights into their functions in vivo, and suggested mechanisms by which lineage selection may be achieved. PMID- 15826836 TI - ETV6-NTRK3: a chimeric protein tyrosine kinase with transformation activity in multiple cell lineages. AB - The ETV6-NTRK3 (TEL-TRKC) gene fusion was discovered by breakpoint analysis of the t(12;15)(p13;q25) translocation associated with congenital fibrosarcoma, a pediatric soft tissue malignancy. ETV6-NTRK3 (EN) encodes the sterile alpha motif oligomerization domain of the ETV6 (TEL) transcription factor linked to the protein tyrosine kinase domain of the neurotrophin-3 receptor NTRK3 (TRKC). The EN chimeric oncoprotein links to multiple signaling cascades including Ras-MAP kinase and PI3K-AKT through the IRS-1 adapter protein. Recent evidence indicates that a functional insulin-like growth factor 1 receptor axis and higher order polymer formation are essential for EN oncogenesis. EN has been detected in other malignancies, including secretory breast carcinoma. This chimeric oncoprotein is therefore unique in being expressed in tumors derived from multiple cell lineages. PMID- 15826837 TI - Fusion oncogenes and tumor type specificity--insights from salivary gland tumors. AB - Salivary gland tumors are frequently characterized by recurrent chromosome translocations, which have recently been shown to result in pathogenetically relevant fusion oncogenes. These genes encode novel fusion proteins as well as ectopically expressed normal or truncated proteins, and are found in both benign and malignant salivary gland tumors. The major targets of the translocations are DNA-binding transcription factors (PLAG1 and HMGA2) involved in growth factor signaling and cell cycle regulation, and coactivators of the Notch (MAML2) and cAMP (TORC1) signaling pathways. Identification of these fusion oncogenes has contributed to our knowledge of molecular pathways leading to epithelial tumors in general, and to salivary gland tumors in particular. Interestingly, the fusions in salivary gland tumors do not seem to be as tumor type specific as those in leukemias and sarcomas. Instead, they may function by activating basic transformation pathways that can function in multiple cell types. The downstream gene products of these fusions will be important targets for development of new intracellular therapeutic strategies. PMID- 15826838 TI - Fusion oncogenes in tumor development. AB - Currently, all identified fusion oncogenes are found in rare tumor forms, and most of them only in specific tumor types. Some fusion oncogenes are frequent in healthy individuals suggesting that they rarely induce tumor growth. Multiple double-strand breaks that cluster in time and space increases the risk for formation of fusion oncogenes genes. The normal cell type specific spatial distribution of chromatin and genes in interphase nuclei may affect the risk for fusion of specific genes. Transcriptional orientation, splicing of reading frames, size and sequences of breakpoint introns are other risk factors. The biological activity of fusion oncoproteins is the most important factor for penetrance. The effects in specific target cells may explain the tumor type specificity of most fusion oncogenes. PMID- 15826839 TI - Differential scanning calorimetric studies of a Bacillus halodurans alpha amylase. AB - The thermal unfolding of Amy 34, a recombinant alpha-amylase from Bacillus halodurans, has been investigated using differential scanning calorimetry (DSC). The denaturation of Amy 34 involves irreversible processes with an apparent denaturation temperature (T(m)) of 70.8 degrees C at pH 9.0, with four transitions, as determined using multiple Gaussian curves. The T(m) increased by 5 degrees C in the presence of 100-fold molar excess of CaCl2 while the aggregation of Amy 34 was observed in the presence of 1000-fold molar excess of CaCl2. Increase in the calcium ion concentration from 1- to 5-fold molar excess resulted in an increase in calorimetric enthalpy (DeltaH(cal)), however, at higher concentrations of CaCl2 (up to 100-fold), DeltaH(cal) was found to decrease, accompanied by a decrease in entropy change (DeltaS), while the T(m) steadily increased. The presence of 100-fold excess of metal chelator, EDTA, resulted in a decrease in T(m) by 10.4 degrees C. T(m) was also decreased to 61.1 degrees C and 65.9 degrees C at pH 6.0 and pH 11.0, respectively. PMID- 15826840 TI - The utility of neurophysiological markers in the study of alcoholism. AB - OBJECTIVE: This review attempts to differentiate neuroelectric measures (electroencephalogram (EEG), event-related potentials (ERPs) and event-related oscillations (EROs)) related to acute and chronic effects of alcohol on the brain from those that reflect underlying deficits related to the predisposition to develop alcoholism and related disorders. The utility of these neuroelectric measures as endophenotypes for psychiatric genetics is evaluated. METHODS: This article reviews the main findings of EEG and ERP abnormalities in alcoholics, offspring of alcoholics at high risk to develop alcoholism and the electrophysiological effects of alcohol on high risk compared to low-risk offspring. It highlights findings using EROs, a fast developing tool in examining brain function and cognition. It also reviews evidence of genetic findings related to these electrophysiological measures and their relationship to clinical diagnosis. RESULTS: Many of these abnormal neuroelectric measures are under genetic control, may precede the development of alcoholism, and may be markers of a predisposition toward the development of a spectrum of disinhibitory conditions including alcoholism. Genetic loci underlying some neuroelectic measures that involve neurotransmitter systems of the brain have been identified. CONCLUSIONS: Quantitative neuroelectric measures (EEG, ERPs, EROs) provide valuable endophenotypes in the study of genetic risk to develop alcoholism and related disorders. SIGNIFICANCE: Genetic studies of neuroelectric endophenotypes offer a powerful strategy for identifying susceptibility genes for developing psychiatric disorders, and provide novel insights into etiological factors. PMID- 15826841 TI - Lack of asymmetrical transfer for linguistic stimuli in schizophrenia: an ERP study. AB - OBJECTIVE: To assess the mechanisms underlying lack of speeded information transfer asymmetry (faster right to left) for verbal information in schizophrenia. METHODS: Interhemispheric transfer times (IHTT) between the hemispheres were assessed using a lateralized lexical-decision task in males with schizophrenia (N = 12) and matched controls (N = 12). Words were presented to the left visual field (LVF), right visual field (RVF), or bilaterally (BVF) while 128 channel EEG was recorded continuously. A direct measure of IHTT in each direction was obtained by comparing the latencies of the N160 evoked potential (EP) component in the hemispheres contralateral and ipsilateral to stimulation. RESULTS: Controls showed faster information transfer from the right to left hemisphere (R-to-L) for linguistic stimuli. The two groups did not differ for IHTTs L-to-R. Lack of IHTT asymmetry in the schizophrenia groups was associated with an overall concomitant decrease in the amplitude of the N160 in the right hemisphere. CONCLUSIONS: Differences in IHTT asymmetry may be attributed to lack of right hemisphere activation and not callosal dysfunction as has been previously suggested. SIGNIFICANCE: It is suggested that a relative excess of myelinated axons in the right hemisphere speeds IHTT faster R-to-L, findings are discussed with reference to differences in right hemisphere white matter connectivity in schizophrenia. PMID- 15826842 TI - Acute isovolemic anemia impairs central processing as determined by P300 latency. AB - OBJECTIVE: Acute anemia slows the responses to clinical tests of cognitive function. We tested the hypothesis that these slowed responses during acute severe isovolemic anemia in healthy unmedicated humans result from impaired central processing. METHODS: A blinded operator measured the latency of the P300 peak in nine healthy volunteers at each volunteer's baseline hemoglobin concentration (Hb), and again after isovolemic hemodilution to Hb 5 g/dL. At both Hb concentrations, the P300 latency was measured twice: with the blinded subject breathing air or 100% oxygen, administered in random order. RESULTS: Anemia increased P300 latency significantly from baseline values (P < 0.05). Breathing oxygen during induced anemia resulted in a P300 latency not different from that at baseline when breathing air (P = 0.5) or oxygen (P = 0.8). CONCLUSIONS: Impaired central processing is, at least in part, responsible for the slowed responses and deficits of cognitive function that occur during acute isovolemic anemia at Hb 5-6 g/dL. SIGNIFICANCE: The P300 latency appears to be a potential measure of inadequate central oxygenation. In healthy young adults with acute anemia, erythrocytes should be transfused to produce Hb>5-6 g/dL. As a temporizing measure, administration of oxygen can reverse the cognitive deficits and impaired central processing associated with acute anemia. PMID- 15826843 TI - Examining the diagnostic utility of EEG power measures in children with attention deficit/hyperactivity disorder. AB - OBJECTIVE: This study investigated the diagnostic utility of EEG power during eyes-closed resting conditions for children with attention-deficit/hyperactivity disorder (AD/HD). METHODS: Subjects consisted of 253 boys with AD/HD combined type and 67 age-matched controls. EEG was recorded from 21 sites during an eyes closed resting condition and was Fourier transformed to provide estimates for total power and absolute and relative power in delta, theta, alpha and beta bands. Factor analysis was used to group sites into frontal, central and posterior regions, with these data subjected to cluster analysis. Logistic regression was performed on the entire AD/HD sample versus control, for AD/HD clusters versus control, and then for each AD/HD cluster independently versus control, using total, absolute and relative power measures. RESULTS: Logistic regression performed on the clusters independently produced the best classification results, with a sensitivity of 89.0% and a specificity of 79.6%, with an overall classification accuracy of 87.0%. CONCLUSIONS: The obtained classification results are supportive of an independent diagnostic test for AD/HD based on EEG power at rest. SIGNIFICANCE: This is the first study to investigate sensitivity and specificity of EEG power for AD/HD in a resting condition. PMID- 15826844 TI - Lateralization of cerebral hemodynamics during Wisconsin Card Sorting Test: a functional transcranial Doppler sonography study. AB - OBJECTIVE: Studies on lateralization of cerebral metabolism during Wisconsin Card Sorting Test (WCST), a well-known paradigm of category learning, have shown mixed results. Moreover, sorting dimension (number, color and shape) is a cofounder of laterality in WCST. Functional transcranial Doppler sonography (fTCD) has a high temporal resolution and allows the measurement of mean cerebral blood flow velocity (CBFV) in the middle (MCA) and anterior cerebral arteries (ACA), which supply lateral and medial parts of the frontal and parietal lobes, respectively. We used fTCD to investigate CBFV changes occurring in both MCA and ACA during WCST and different sorting dimensions. METHODS: Twenty-one subjects underwent twice two distinct phases of the WCST, namely maintaining a rule (maintaining set) and searching for a new rule (set shifting), during bilateral fTCD of the MCA and ACA. RESULTS: There was a left-sided dominance of CBFV during maintaining set and set shifting in the MCA. CBFV was not associated with test performance. The sorting dimension number caused the highest CBFV increase in both MCA and ACA during maintaining set, and the sorting dimension shape caused lowest CBFV decrease in both MCA during set shifting. CONCLUSIONS: This study confirms results that cerebral blood flow (CBF) lateralizes to the left side during WCST. The 3 sorting dimensions provoked distinct processing speed during maintaining set and set shifting, but caused no effect on hemispheric lateralization. SIGNIFICANCE: Functional transcranial Doppler sonography can be used to assess CBFV during WCST and different sorting dimensions, and the latter modulate reaction time and cerebral hemodynamics. PMID- 15826846 TI - Repetitive transcranial magnetic stimulation (rTMS) at high and low frequency: an efficacious therapy for major drug-resistant depression? AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) is proposed for the treatment of drug-resistant depression. Studies performed in accordance with evidence-based medicine (EBM) are scarce, particularly in seeking optimal treatment and evaluation parameters. We aimed to test various types of rTMS in a large sample of depressed patients following EBM rules and to investigate treatment-related changes in plasma levels of neurotransmitters involved in depression. METHODS: Seventy-one drug-resistant depressed patients were randomly assigned to low (1 Hz) or high (17 Hz) rate TMS, applied for 5 days over the left dorsolateral prefrontal cortex (L-DLPFC). Patients were separated into two study designs. One group (20 patients) received only active treatment, while the other entered a double-blind, placebo-controlled, crossover design. Pre- and post treatment blood samples were taken for evaluation of plasma levels of dopamine and serotonin. RESULTS: After a week of treatment patients had a measurable benefit. However, overall the placebo stimulation did not differ significantly from real stimulation, nor were differences observed between the two rates of rTMS. The only difference emerged when the real stimulation was applied at 17 Hz following placebo treatment. Plasma levels of neurotransmitters between active and placebo rTMS were similar. CONCLUSIONS: Using the treatment schedule of 1 week, although a clinical improvement after active treatment was indeed observed, this was both clinically and biochemically indistinguishable from that seen in the placebo arm. SIGNIFICANCE: This suggests that most of the previous emphasis, for short period of treatment, should be tempered down and that further work is required in order to verify whether optimal stimulation and evaluation parameters for TMS-treatment of depression beyond the placebo effect may be found following EBM rules. PMID- 15826847 TI - Short-lasting impairment of temperature perception by high frequency rTMS of the sensorimotor cortex. AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) has become a useful tool for investigating and even modulating human brain function. RTMS of the human motor cortex can produce changes in excitability that outlast the period of stimulation. To investigate the persistent effect of high-frequency rTMS of sensorimotor cortex (SM1) on somatosensory function. METHODS: We evaluated the thermal thresholds (cold and warm sensation) in 14 normal subjects before and after a short train of 5Hz rTMS over the SM1 or occipital cortex (OC). RESULTS: Threshold for cold perception was increased immediately after rTMS of the left SM1 and no effects at all were noticed after OC stimulation. There was a slight, not significant, increase of warm threshold immediately after the rTMS of the left SM1 and no effects at all were noticed after OC stimulation. CONCLUSIONS: High frequency rTMS over primary sensorimotor cortex seems to modulate sensory function related to thermal (cold) perception. SIGNIFICANCE: The method may be useful for both the study of normal human physiology of temperature perception and for rTMS based manipulation of brain plasticity in patients with sensory disturbances. PMID- 15826845 TI - Spatial-anatomical mapping of NoGo-P3 in the offspring of alcoholics: evidence of cognitive and neural disinhibition as a risk for alcoholism. AB - OBJECTIVE: The concept of disinhibition as a behavioral and biological trait has been considered to be involved in the etiology of alcoholism and its co-existing disorders. The magnitude and functional mapping of event-related potential P3(00) components were analyzed, in order to examine the possible response inhibition deficits in the offspring of alcoholics. METHODS: The P3 components were compared between 50 offspring of alcoholics (OA) and a matched normal control group (NC) using a visual Go/NoGo task. The low-resolution electromagnetic tomography (LORETA) was used to analyze the functional brain mapping between groups. RESULTS: The results indicated that the OA group manifested decreased P3 amplitude during the NoGo but not the Go condition compared to the NC group. The voxel-by-voxel analysis in LORETA showed group differences at several brain regions including prefrontal areas during the processing of NoGo but not Go signals. CONCLUSIONS: The decreased NoGo-P3 suggests that cognitive and neural disinhibition in offspring of alcoholics may serve as a neurocognitive index for a phenotypic marker in the development of alcoholism and related disorders. SIGNIFICANCE: Dysfunctional neural and response inhibition in the offspring of alcoholics perhaps provides an endophenotypic marker of risk for the development of alcoholism and related disorders. PMID- 15826848 TI - Attention induces reciprocal activity in the human somatosensory cortex enhancing relevant- and suppressing irrelevant inputs from fingers. AB - OBJECTIVE: We studied whether attention regulates information processing in the human primary somatosensory cortex (SI) by selective enhancement of relevant- and suppression of irrelevant information. METHODS: Under successive and simultaneous electric stimuli to both the right index and middle fingers, tactile stimuli were randomly (20%) presented on one of the two fingers in separate two runs exchanging the finger. Subjects were requested to discriminate the tactile stimuli in an attention task to induce attention to one finger and to ignore the stimuli in a control task to avoid such an attention focus. Somatosensory evoked magnetic fields were measured only for the two-finger electric stimulation and an early component (M50) was analyzed. RESULTS: In spite of the two-finger simultaneous stimulation, attention to either the index or middle finger lowered or heightened the M50-sourse location, respectively. The attention task did not increase the M50 amplitude. CONCLUSIONS: Attention to a finger enhanced selectively the representation of the finger in the SI cortex. However, this SI activity did not increase the M50 amplitude, suggesting that the attention suppressed another finger region receiving the unattended inputs. SIGNIFICANCE: Attention regulates the SI activity by selectively enhancing the task-relevant information and by filtering out other noise inputs. PMID- 15826849 TI - Coherence of the electroencephalogram during the first sleep cycle. AB - OBJECTIVE: The increasing amplitude of the electroencephalogram (EEG) during non rapid eye movement sleep implies a progressive synchronization of neuronal activity. We sought to characterize the spatial relationship of cortical activity at different frequencies during the first sleep cycle, focusing on sleep stages 3 and 4 (slow wave sleep). METHODS: Sleep EEGs were obtained at home from six adults using a portable recorder. Signal power and magnitude squared coherence were measured during the first sleep cycle. Spectra obtained from bipolar and common reference derivations were compared. RESULTS: During slow wave sleep, signal power is highest in the delta frequency band and regional coherence below 5 Hz is broadly distributed. Although signal power in the alpha and sigma frequency bands is lower, peaks of regional coherence in those bands are similar to or higher than delta-band coherence. Regional coherence during slow wave sleep is differentially distributed with a 14 Hz component in central and posterior regions and a 10 Hz component in frontal and central regions. CONCLUSIONS: Ten and 14 Hz rhythms are an essential component of slow wave sleep. SIGNIFICANCE: The interpretation of scalp EEG power and coherence spectra is limited by the lack of a satisfactory recording reference. However, conclusions can be made by comparing and contrasting results from both bipolar and common reference recordings. PMID- 15826850 TI - Cardiovascular variability during periodic leg movements: a spectral analysis approach. AB - OBJECTIVE: Changes in cardiovascular measures have been advocated as sensitive markers of phasic events arising from sleep. The current study was aimed to analyse the effects of periodic leg movements (PLMS) on heart rate variability (HRV) during NREM sleep in patients having restless legs syndrome and periodic leg movements during sleep. METHODS: The absolute and normalized high- and low frequency peaks from spectral analysis (FFT) of R-R intervals, the HRV changes using wavelet transform, the geometric and time domain HRV were measured in 14 patients with restless legs syndrome and PLMS. The analysis was done comparing one hundred, 10 min periods with PLMS (PLMS+) and 60 periods without PLMS (PLMS-) in stage 2 of NREM sleep. In 8 patients analysis was also done in slow wave sleep (SWS). RESULTS: Occurrence of PLMS induced changes in geometrical indices of HRV, with a rise of the triangular index and the triangular interpolation of R-R intervals in PLMS+ periods (P < 0.0001). Small changes in time domain indices were found during PLMS+ periods, while the SD of the R-R intervals (SDNN), reflecting global HRV, was significantly higher (P = 0.001). While the low frequency (LF) power significantly increased in PLMS+ periods (P < 0.0001), high frequency (HF) power showed a weak and not significant reduction. The rise in sympathetic activity as detected by frequency domain HRV analysis was related to density and interval of PLMS. Comparison between sleep stages of NREM sleep demonstrated lower values of HRV measures when PLMS+ periods occur in SWS. CONCLUSIONS: Overall, PLMS occurrence was associated with a shift to increased sympathetic activity without significant changes in cardiac parasympathetic activity. The frequency domain analysis of HRV appears to be an easy tool to estimate the autonomic changes related to PLMS and PLMS- arousals and to differentiate their occurrence during stage 2 and deep sleep. SIGNIFICANCE: Spectral HRV measures may offer a simple approach to estimate the degree of autonomic changes occurring in relation to periodic leg movements in restless legs patients. PMID- 15826851 TI - Cortical excitability during prolonged antiepileptic drug treatment and drug withdrawal. AB - OBJECTIVE: Previous reports characterized the effects of administration of single oral doses of antiepileptic drugs (AED) on cortical excitability. However, AED effects on cortical excitability, and their relationship to plasma blood levels, during chronic drug administration at therapeutic doses are not known. The objective of the study was to determine whether plasma blood levels during chronic administration at therapeutic doses would accurately predict changes in corticomotor excitability. METHODS: We used transcranial magnetic stimulation (TMS) to measure cortical excitability during 5 weeks administration of carbamazepine (CBZ) and lamotrigine (LTG), and subsequent AED withdrawal in 20 healthy volunteers. Data were analyzed using ANOVA(RM) and regression analysis. RESULTS: Resting motor thresholds (r-MT) increased with increasing total and free CBZ and LTG levels during drug administration, but not drug withdrawal. After acute AED withdrawal, r-MT elevation persisted in most individuals with CBZ despite undetectable plasma levels, compared to a rapid normalization with LTG. In contrast, acute drug withdrawal resulted in a transient decrease in r-MT in 3/10 individuals with CBZ and 2/10 with LTG. CONCLUSIONS: Plasma levels provide information on motor cortical function during active treatment phases but not during AED withdrawal. SIGNIFICANCE: The transient decrease in r-MT associated with acute AED withdrawal could represent a physiological substrate contributing to AED withdrawal seizures. PMID- 15826852 TI - Hidden focal EEG seizures during prolonged suppressions and high-amplitude bursts in early infantile epileptic encephalopathy. AB - OBJECTIVE: We report on a 27-month-old female with atypical early infantile epileptic encephalopathy (EIEE), who developed tonic spasms, partial seizures and myoclonic jerks along with episodic bradycardia at 5 days. METHODS: We recorded digital electroencephalography (EEG) using either an 11-channel neonatal montage or 19 channel scalp electrodes, at 200 Hz sampling rate, and a single reference for a minimum of 30 min. RESULTS: At 18 days EEG showed suppression-burst (SB) patterns during wakefulness and sleep. Tonic spasms concomitant with bursts recorded as brief, low-amplitude fast waves. EEG at 8 months showed increased amplitude of bursts to 1 mV and extension of suppression periods to 65 s. By increasing recording sensitivity, we detected focal epileptiform discharges of slow rhythmic sharp and slow waves building to 30 microV during suppression periods. Status epilepticus occurred at 16 months. EEG at 27 months returned to the previous SB pattern with rare partial seizures. CONCLUSIONS: This report is the first to demonstrate clinically silent focal EEG seizures during prolonged suppression periods in atypical EIEE by off-line digital EEG. SIGNIFICANCE: Digital EEG sensitivity can reveal covert electrical activity during suppression periods in epileptic neonates and infants. PMID- 15826853 TI - Origin, structure, and role of background EEG activity. Part 3. Neural frame classification. AB - OBJECTIVE: To show that cortical responses to conditioned stimuli (CS) include intermittently induced spatial patterns of amplitude modulation (AM) of beta gamma oscillation called frames. METHODS: EEGs were recorded from 8x8 high density arrays fixed on primary sensory cortices of rabbits trained to discriminate CS with reinforcement (CS+) from those without (CS-). EEG frames were located with a pragmatic information index, H(e). The spatial patterns of the first 3 frames on each of 37-40 trials were measured by the square of 64 analytic amplitudes from the Hilbert transform to give points in 64-space. The questions were asked: Did the frames from CS+ trials and CS- trials differ within each sequential group? Did the 3 frames differ from each other (form 3 clusters of points)? RESULTS: EEG frames that were identified by high H(e) had AM patterns that could be classified with respect to CS+ and CS- well above chance levels. Two stages of correct frame classification occurred on each trial: 40-130 ms after CS onset with a gamma carrier frequency, and 450-550 ms with a beta carrier frequency. Peak power in the beta frames was double that in gamma frames, and mean pattern surface area of beta frames was nearly 4-fold greater. CONCLUSIONS: Under the impact of a CS on a sensory neocortex, the background EEG activity reorganized in sequential frames of coordinated activity, first local and modality-specific, thereafter global. SIGNIFICANCE: The size, texture and duration of these AM patterns indicate that spatial patterns of human beta frames may be accessible with high-density scalp arrays for correlation with phenomenological reports by human subjects. PMID- 15826854 TI - Geodesic photogrammetry for localizing sensor positions in dense-array EEG. AB - OBJECTIVE: An important goal for functional brain studies using EEG technology is to estimate the location of brain sources that produce the scalp-recorded signals. The accuracy of source estimates is dependent upon many variables, one of which is the accurate description of the scalp positions of the EEG sensors. The objective of the present research was to develop a photogrammatic method for sensor localization that is fast, accurate, and easy to use. METHODS: With the novel photogrammetric method, multiple cameras were arranged in a geodesic array, and images of the sensors on the subject's head were acquired allowing for the reconstruction of the 3D sensor positions. RESULTS: Data from the photogrammetric method were compared with data acquired with the conventional electromagnetic method. The accuracy of the photogrammatic method, quantified as RMS of the measured positions and the actual known positions, was similar (mean error = 1.27 mm) to the electromagnetic method (mean error = 1.02 mm), and both approximated the localization error of the calibration object (mean error = 0.56 mm). CONCLUSIONS: Accurate determination of 3D sensor positions can be accomplished with minimal demands on the time of the subject and the experimenter using the photogrammetric method. PMID- 15826855 TI - Evaluation of the distortion of EEG signals caused by a hole in the skull mimicking the fontanel in the skull of human neonates. AB - OBJECTIVE: Interpretation of Electroencephalography (EEG) signals from newborns is in some cases difficult because the fontanels and open sutures produce inhomogeneity in skull conductivity. We experimentally determined how EEG is influenced by a hole mimicking the anterior fontanel since distortion of EEG signals is important in neurological examinations during the perinatal period. METHODS: Experiments were carried out on 10 anesthetized farm swine. The fontanel was mimicked by a hole (12 x 12 mm) in the skull. The hole was filled with 3 types of medium differing in conductivity (air, 0 S/m; sucrose-agar, 0.017 S/m; saline-agar, 1.28 S/m). Three positions of the snout were stimulated with a concentric bipolar electrode to activate cortical areas near the middle, the edge, and the outside of the hole. The somatic-evoked potential (SEP) was recorded by a 4 x 4 electrode array with a 4mm grid spacing. It was placed on the 4 quadrants of a 28 x 28 mm measurement area on a saline-soaked filter paper over the skull, which served as artificial scalp. RESULTS: The SEP over the hole was clearly stronger when the hole was filled with sucrose- or saline-agar as compared to air, although paradoxically the leakage current was stronger for the sucrose- than saline-agar. The current leaking from the hole was strongly related to position of the active tissue. It was nearly negligible for sources 6-10 mm away from the border of the hole. The distortion was different for 3 components of the SEP elicited by each stimulus, probably indicating effects of source distance relative to the hole. CONCLUSIONS: EEG is strongly distorted by the presence of a hole/fontanel with the distortion specifically dependent on both conductivity of the hole and source location. SIGNIFICANCE: The distortion of the EEG is in contrast to the lack of distortion of magnetoencephalography (MEG) signals shown by previous studies. In studying brain development with EEG, the infant's head and sources should be modeled accurately in order to relate the signals to the underlying activity. MEG may be particularly advantageous over EEG for studying brain functions in infants since it is relatively insensitive to skull defects. PMID- 15826856 TI - Differences in potentials and excitability properties in simulated cases of demyelinating neuropathies. Part I. AB - OBJECTIVE: The aim of this study is to investigate the potentials (intracellular, extracellular, electrotonic) and excitability properties (strength-duration and charge-duration curves, strength-duration time constants, rheobases, recovery cycles) in three cases of uniform myelin wrap reduction (20, 50 and 70%) along the fibre length. METHODS: The internodally systematically demyelinated cases (termed as ISD1, ISD2 and ISD3) are simulated using our previous double cable model of human motor fibres. RESULTS: In the more severely demyelinated cases, the intracellular potentials are with significantly reduced amplitude, prolonged duration and slowed conduction velocity, whereas the electrotonic potentials show greater increase in the early part of the hyperpolarizing responses. The radial decline of the extracellular potential amplitudes depends on the radial distance of the field point and increases with the increase of the distance and demyelination. The time constants and rheobasic currents increase with the increase of the degree of demyelination. In the recovery cycles, the more severely demyelinated cases have greater refractoriness (the increase in threshold current during the relative refractory period), supernormality and less late subnormality than the normal case. CONCLUSIONS: The myelin thickness has significant effects on the potentials and axonal excitability properties of the simulated demyelinated human motor fibres. The obtained abnormalities in the potentials and excitability properties can be observed in Charcot-Marie-Tooth disease type 1A (CMT1A). SIGNIFICANCE: The study provides new information about the pathophysiology of human demyelinating neuropathies. PMID- 15826857 TI - Differences in potentials and excitability properties in simulated cases of demyelinating neuropathies. Part II. Paranodal demyelination. AB - OBJECTIVE: The purpose of the present investigation is to study the potentials and axonal excitability properties in progressively greater degrees of uniform paranodal demyelination of human motor nerve fibres. METHODS: Using our previous double cable model of human motor nerve fibre, 3 paranodally systematically demyelinated cases (termed as PSD1, PSD2 and PSD3) are simulated by an uniform paranodal resistance reduction (20, 50 and 77%) along the fibre length. RESULTS: Considerably reduced amplitudes, prolonged durations and slowed conduction velocities are obtained for the intracellular potentials of the PSD2 and PSD3 cases. In contrast, the electrotonic potentials show abnormally greater increase in the early part of the hyperpolarizing responses. The extracellular potentials indicate increased polyphasia in the PSD3 case. The strength-duration time constants are shorter and the rheobases higher in the demyelinated cases. In the recovery cycles, the demyelinated cases have less refractoriness, greater supernormality and less late subnormality than the normal case. CONCLUSIONS: The reduction of the paranodal seal resistance has significant effects on the potentials and axonal excitability properties of the simulated demyelinated human motor fibres. The obtained abnormalities in the potentials and excitability properties can be observed in vivo in patients with chronic inflammatory demyelinating polyneuropathy. SIGNIFICANCE: The study provides important information about the pathology of human demyelinating neuropathies. PMID- 15826858 TI - MG and LEMS overlap syndrome: case report with electrophysiological and immunological evidence. AB - OBJECTIVE: To report the clinical, electrophysiological, and immunological findings in one patient with MG and LEMS overlap syndrome (myasthenia gravis and Lambert-Eaton myasthenic syndrome). METHODS: Evaluation of clinical, electrophysiological, and immunological findings in one patient with this disease. RESULTS: A female patient with pernicious anemia had clinical findings of ptosis, diplopia, proximal leg weakness, areflexia and a positive edrophonium test as well as the classic patterns initially of MG and later of LEMS on successive repetitive nerve stimulation (RNS) tests. Immunologically she demonstrated positive acetylcholine receptor antibody (ACh-ab) and positive N type voltage-gated calcium-channel antibody (VGCC-ab) titers. CONCLUSIONS: Among five reported cases combining the features of MG and LEMS and having positive AChR- and VGCC-ab titers, our patient provides the most convincing clinical, electrophysiological, and immunological evidence for the existence of MG and LEMS overlap syndrome. SIGNIFICANCE: MG and LEMS overlap syndrome is a distinct entity. PMID- 15826859 TI - Decrement of the skin conductance response to repeated volitional inspiration. AB - OBJECTIVE: To examine response decrement of the recently reported inspiratory skin conductance response (SCR) [Lim CL, Seto-Poon M, Clouston PD, Morris JG. Sudomotor nerve conduction velocity and central processing time of the skin conductance response. Clin Neurophysiol 2003;114:2172-80]. METHODS: Twelve healthy adult volunteers performed 3 tasks (A) a control task of maintaining tidal breathing and then two randomized tasks, (B) a deep inspiration to a target oral pressure and (C) tapping with a finger. Each task was performed 30 times on cue every 20s in 3 runs with 5 min of rest between runs. The SCR, oral pressure, airflow, inspired volume and cue signal were recorded continuously and analysed offline. SCR amplitude was logarithmically transformed and then statistically analysed, using a linear mixed effects model, as a function of run number, trial number and absolute error between target and actual oral pressures. RESULTS: Inspiratory efforts elicited exponentially decreasing SCR amplitude with increasing trial number during each run (P < 0.0001). After adjusting for trial number, the mean SCR amplitude of the second and the third run were, respectively, 24.2 (95% CI (0.175, 0.336), P < 0.001) and 14.4% (95% CI (0.104, 0.200), P < 0.001) of the first run amplitude. CONCLUSIONS: Volitional deep inspiration reliably activates an SCR that exhibits response decrement with repetition, which may be habituation. SIGNIFICANCE: The volitional inspiratory SCR may assist in the assessment of sympathetic autonomic status in patients with peripheral afferent neuropathy. PMID- 15826860 TI - Axonal potassium conductance and glycemic control in human diabetic nerves. AB - OBJECTIVE: To investigate the effects of hyperglycemia on axonal excitability and potassium conductance in human diabetic nerves. METHODS: Threshold tracking was used to measure excitability indices, which depend on potassium channels (supernormality, late subnormality, threshold electrotonus, and a current/threshold relationship) in median motor axons of 96 diabetic patients. The effects of hyperglycemia on these indices were analyzed. RESULTS: Among diabetic patients, higher serum hemoglobin A1c (HbA1c) levels were significantly associated with greater supernormality (P = 0.04) and smaller late subnormality (P = 0.02), suggestive of reduced nodal/paranodal potassium currents under hyperglycemia. Threshold electrotonus and current/threshold relationships did not correlate with HbA1c levels, but partly related with nerve conduction slowing. CONCLUSIONS: Hyperglycemia could reduce nodal potassium conductances, possibly due to reduced membranous potassium gradient or suppression of potassium channels. In contrast, internodal potassium conductances may be determined by both metabolic factors and structural changes such as exposure of internodal channels by demyelination. SIGNIFICANCE: Measurements of the excitability indices could provide new insights into nodal and internodal axonal membrane properties in human diabetic neuropathy, whereas multiple factors can affect especially internodal properties. PMID- 15826861 TI - Variability in spatial normalization of pediatric and adult brain images. AB - OBJECTIVE: Normalization of brain images is a necessity for group comparisons of source analyses based on realistic head models. In this paper we compared the outcome of a linear registration method for brain images of psychiatric and control groups of different ages in order to assess the relative adequacy of normalization in such diverse groups. METHODS: Magnetic Resonance images (MRI) of the brains of pediatric and adolescent subjects (mean ages 19 and 10.5 years) with a pervasive developmental disorder (PDD) and their healthy controls were included. A simple voxel-wise test of the group variances in image intensities was performed to evaluate regional differences in registration quality. Dipole analysis of visual P1 was performed to establish whether source locations were comparable across groups. RESULTS: Significant differences between pediatric groups were found in white matter and thalamic regions of the brain. For all other group-wise comparisons, differences were confined to skull and neck regions. Dipole locations were found to be more anteriorly located in the adolescent groups. CONCLUSIONS: The normalization procedure used in this paper is based on a brain template of normal adult brains from a restricted age group, and the results show that the use of this method in pediatric groups is less adequate. The method seems suitable for use in psychiatric groups. Also, the generators of visual P1 in PDD patients were found to be comparable to controls. SIGNIFICANCE: The results suggest that this existing normalization method can be used in diverse populations, but is less suitable for pediatric images. PMID- 15826862 TI - Effect of spatial attention on human sensorimotor integration studied by transcranial magnetic stimulation. AB - OBJECTIVE: Recent transcranial magnetic stimulation (TMS) studies showed that the sensory input can decrease the motor cortex excitability (afferent inhibition). To clarify the effect of attention on sensorimotor integration, we investigated the effect of spatial attention on afferent inhibition. METHODS: Right median nerve electrical stimulation followed, at variable delays (10-300 ms), by TMS over the left motor cortex was applied to 9 subjects, during 3 conditions; spatial attention to the right and left hand, and control (no attention) tasks. RESULTS: Inhibition of the motor evoked potential occurred at inter-stimulus interval of 20 and 100 ms, which was more was marked during spatial attention to the right than to the left hand. CONCLUSIONS: Enhancement of the afferent inhibition induced by spatial attention to the stimulated side is likely to reflect the interaction between attention and sensorimotor integration. SIGNIFICANCE: The spatial attention may modulate the sensorimotor integration studied by afferent inhibition of the MEP. PMID- 15826863 TI - Temporal activation pattern of parietal and premotor areas related to praxis movements. AB - OBJECTIVE: We sought to determine the cortical physiology underlying praxis movements in normal subjects using electroencephalography (EEG). METHODS: Eight normal subjects were instructed to perform six types of self-paced tool-use pantomime and communicative gesture movements with the right hand. We recorded 64 channel EEG using a linked ear reference and electromyogram (EMG) from right thumb and forearm flexors. RESULTS: Data revealed early slow wave components of the movement-related cortical potential (MRCP) beginning over the left parietal area about 3s before movement onset, similarly for both movement types. At movement onset, maximal amplitude was present over central and bilateral sensorimotor areas. Event-related desynchronization (ERD) in the beta band was seen over the left parietal and sensorimotor cortices during preparation, later spreading to the homologous area of the right hemisphere. Alpha ERD was mainly in the left sensorimotor cortex about 1.5s before movement onset. Beta ERD in mesial frontal areas was greater during preparation for tool use compared to communicative gesture movements. Mesial frontal beta event-related synchronization (ERS) developed more rapidly after communicative gestures than tool-use. CONCLUSIONS: The dynamics of parietal and frontal activities indicates the timing of these areas in the production of praxis. The posterior parietal cortex contributes to the early slow wave negativity of the MRCP. SIGNIFICANCE: Planning self-paced praxis movements begins as early as 3s before movement in the left parietal area and subsequently engages frontal cortical regions. PMID- 15826864 TI - Asymmetric spatiotemporal patterns of event-related desynchronization preceding voluntary sequential finger movements: a high-resolution EEG study. AB - OBJECTIVE: To study spatiotemporal patterns of event-related desynchronization (ERD) preceding voluntary sequential finger movements performed with dominant right hand and nondominant left hand. METHODS: Nine subjects performed self-paced movements consisting of three key strokes with either hand. Subjects randomized the laterality and timing of movements. Electroencephalogram (EEG) was recorded from 122 channels. Reference-free EEG power measurements in the beta band were calculated off-line. RESULTS: During motor preparation (-2 to -0.5s with respect to movement onset), contralateral preponderance of event-related desynchronization (ERD) (lateralized power) was only observed during right hand finger movements, whereas ERD during left hand finger movements was bilateral. CONCLUSIONS: For right-handers, activation on the left hemisphere during left hand movements is greater than that on the right hemisphere during right hand movements. SIGNIFICANCE: We provide further evidence for motor dominance of the left hemisphere in early period of motor preparation for complex sequential finger movements. PMID- 15826865 TI - Periodic motor cortical excitability changes associated with PSDs of EEG in Creutzfeldt-Jakob disease (CJD). AB - OBJECTIVE: To assess the temporal relationship between excitability changes of the motor cortex and periodic synchronous discharges (PSDs) of electroencephalograms (EEGs) in Creutzfeld-Jakob disease (CJD) patients. METHODS: In four patients with classical type CJD we gave transcranial magnetic stimulation (TMS) randomly in time while simultaneously recording their EEGs and electromyograms (EMGs). The intensity of TMS was fixed to evoke 0.3-0.5 mV EMG in the abductor pollicis brevis muscle (APB) during periods in which there were no PSDs. We assessed the relation between the timing of TMS and EMG response sizes when PSD appeared in EEGs. The time interval between TMS and the onset of preceding PSD was plotted against the amplitude of acquired motor evoked potentials (MEPs). RESULTS: The amplitude of MEP was at a minimum when the TMS pulse was given just after the beginning of PSD and was maximal when given just before the onset of the next PSD. As a result, the size of MEP amplitude periodically changed and was phase-locked to the PSD, though its cycle slightly preceded that of PSD. CONCLUSIONS: Our study has demonstrated that excitability changes in motor cortex are closely related to the occurrence of PSD. SIGNIFICANCE: It has been revealed for the first time that motor cortical excitability is related to PSD cycle. PMID- 15826866 TI - Temporal excitation properties of paresthesias evoked by thalamic microstimulation. AB - OBJECTIVE: The neuronal elements mediating the effects of deep brain stimulation (DBS) are unknown. The objective was to determine the strength-duration properties of the neuronal elements that mediate paresthesias evoked by thalamic microstimulation. METHODS: The strength-duration properties of the neuronal elements causing paresthesias were measured using intraoperative microstimulation of the human thalamus. The sample included both concordant (reported in the same region as the mapped sensory receptive fields) and discordant paresthesias (reported in a region different than the mapped sensory receptive fields). RESULTS: There were no significant differences between the chronaxies of concordant and discordant paresthesias. There was no significant correlation between chronaxie and rheobase for concordant paresthesias, but a strong negative correlation existed for discordant paresthesias. CONCLUSIONS: Chronaxies did not distinguish the neuronal elements mediating concordant and discordant paresthesias, but correlations between chronaxie and rheobase suggest that concordant paresthesias were produced by activation of local cells while discordant paresthesias were caused by activation of axons of passage. SIGNIFICANCE: The similarity between the strength-duration properties of paresthesias evoked by thalamic stimulation, tremor reduction evoked by thalamic DBS, and EMG responses to thalamic DBS does not mean that these effects are caused by the same neural elements. PMID- 15826867 TI - The mouse vesicular inhibitory amino acid transporter gene: expression during embryogenesis, analysis of its core promoter in neural stem cells and a reconsideration of its alternate splicing. AB - The vesicular inhibitory amino acid transporter, VIAAT (also known as vesicular GABA transporter VGAT) transports GABA or glycine into synaptic vesicles. To initiate an analysis of the expression and regulation of VIAAT during neurogenesis we have cloned and characterized the mouse Viaat gene. We find that the mouse Viaat coding sequence is encoded by two exons spanning 5.3 kb. A survey of expression by whole mount in situ hybridization of mouse embryos indicates that Viaat is activated early in neuron differentiation and is expressed widely within the developing CNS; however, we did not detect expression in the superficial non-neural structures that express the GABA synthase Gad1. Analysis of the Viaat promoter indicates that a minimal promoter region containing a CG rich sequence is sufficient for efficient expression in neural stem and precursor cells. Our analysis of the Viaat sequence and splicing does not support the existence of two Viaat isoforms as previously proposed [Ebihara et al., Brain Res. Mol Brain Res. 110 (2003), 126-139]. Instead, the alternative isoform Viaat a appears to be due to PCR artifacts that have occurred independently in multiple labs. PMID- 15826868 TI - A transmembrane phospholipase D in Phytophthora; a novel PLD subfamily. AB - Phospholipase D (PLD) is a ubiquitous enzyme in eukaryotes that participates in various cellular processes. Its catalytic domain is characterized by two HKD motifs in the C-terminal part. Until now, two subfamilies were recognized based on their N-terminal domain structure. The first has a PX domain in combination with a PH domain and is designated as PXPH-PLD. Members of the second subfamily, named C2-PLD, have a C2 domain and have, so far, only been found in plants. Here we describe a novel PLD subfamily that we identified in Phytophthora, a genus belonging to the class oomycetes and comprising many important plant pathogens. We cloned Pipld1 from Phytophthora infestans and retrieved full-length sequences of its homologues from Phytophthora sojae and Phytophthora ramorum genome databases. Their promoters contain two putative regulatory elements, one of which is highly conserved in all three genes. The three Phytophthora pld1 genes encode nearly identical proteins of around 1807 amino acids, with the two characteristic HKD motifs in the C-terminal part. Homology of the predicted proteins with known PLDs however is restricted to the two catalytic HKD motifs and adjacent domains. In the N-terminal part Phytophthora PLD1 has a PX-like domain, but it lacks a PH domain. Instead the N-terminal region contains five putative membrane spanning domains suggesting that Phytophthora PLD1 is a transmembrane protein. Since Phytophthora PLD1 cannot be categorized in one of the two existing subfamilies we propose to create a novel subfamily named PXTM-PLD. PMID- 15826869 TI - Lack of microbicidal response in human macrophages infected with Parachlamydia acanthamoebae. AB - Parachlamydia acanthamoebae is an obligate intracellular bacterium naturally infecting free-living amoebae. This potential agent of pneumonia resists destruction by human macrophages, inducing their death by apoptosis. However, the strategy used by Parachlamydia to escape the microbicidal effectors of macrophages remains unknown. In this work, we defined the effect of Parachlamydia on the cytokine secretion (measured in culture supernatants by immunoassays), on the oxidative burst (measured using a fluorogenic probe), on the production of nitric oxide (Griess assay), and on transcription of glutaredoxin, tumor necrosis factor alpha (TNF-alpha) and indoleamine 2,3-dioxygenase (IDO). Living Parachlamydia did not induce an oxidative burst, the secretion of cytokines such as IL-6, IL-10 and TNF-alpha, nor the transcription of TNF-alpha in macrophages. However, living Parachlamydia led to increased secretion of IL-1beta and increased transcription of glutaredoxin, an anti-oxidant. The transcription of IDO, an enzyme, which catalyzes decyclization of l-tryptophan, was slightly up regulated. Heat-inactivated Parachlamydia did not induce either an oxidative burst or the production of pro-inflammatory cytokines. In contrast to living bacteria, it had no effect on the IL-1beta release, but it induced IL-10 secretion. In conclusion, after being internalized, Parachlamydia may resist the microbicidal effectors of human macrophages through not inducing oxidative burst and pro-inflammatory cytokine production. PMID- 15826870 TI - Differential turnover of cortical and trabecular bone in transgenic mice overexpressing cathepsin K. AB - Cathepsin K is a major osteoclastic protease. We have recently shown that overexpression of mouse cathepsin K gene in transgenic UTU17 mouse model results in high turnover osteopenia of metaphyseal trabecular bone at the age of 7 months. The present report extends these studies to a systematic analysis of cortical bone in growing and adult mice overexpressing cathepsin K. Mice homozygous for the transgene locus (UTU17+/+) and their control littermates were studied at the age of 1, 3, 7, and 12 months. Bone properties were analyzed using peripheral quantitative computed tomography (pQCT), histomorphometry, histochemistry, radiography, and biomechanical testing. In addition, the levels of biochemical markers of bone turnover were measured in the sera. Unexpectedly, cortical thickness and cortical bone mineral density were increased in the diaphyseal region of growing and adult UTU17+/+ mice. This was associated with an increased number of vascular canals leading to increased cortical porosity in UTU17+/+ mice without changes in the ultimate bending force or stiffness of the bone. In UTU17+/+ mice, osteopenia of metaphyseal trabecular bone was observed already at the age of 1 month. In sera of 1-month-old UTU17+/+ mice, the activity of tartrate-resistant acid phosphatase 5b was decreased and the levels of osteocalcin increased. Our results support the role of cathepsin K as a major proteinase in osteoclastic bone resorption. Excessive production of cathepsin K induced osteopenia of metaphyseal trabecular bone and increased the porosity of diaphyseal cortical bone. The increased cortical thickness and bone mineral density observed in diaphyses of UTU17+/+ mice demonstrate the different nature and reactivity of trabecular and cortical bone in mice. These results suggest that the biomechanical properties of cortical bone are preserved through adaptation as outlined in Wolff's law. PMID- 15826871 TI - Saeng-Ji-Hwang has a protective effect on adriamycin-induced cytotoxicity in cardiac muscle cells. AB - This study examined the effect of Saeng-Ji-Hwang (SJH: Radix Rehmanniae) on cardiac muscle cells. Adriamycin-exposed H9C2 cardiac muscle cells were treated with a water extract of SJH. The adriamycin induced cell death and caspase-3 activation were significantly inhibited by SJH (2 mg/ml), which can be explained by the increase in Bcl-2 expression and the inhibition of Bax expression. Adriamycin reduced the Mn-SOD protein expression level in H9C2 cardiac muscle cells but a SJH treatment partially but significantly reversed this effect. Manganese (Mn)-TBAP or Mn-TMyM--mitochondria-specific SOD mimetic agent--reduced the adriamycin-induced cytotoxicity. It was also shown that SJH inhibits the release of H2O2 and prevents lipid peroxidation in the presence of adriamycin. This study examined the intracellular GSH level, which showed that adriamycin significantly decreased the intracellular GSH level but SJH increased it. BSO, a selective inhibitor of glutamyl cysteinyl ligase, which is a rate-limiting enzyme in GSH synthesis, did not affect the viability of the cardiac muscle cells. However, a combination of BSO with SJH in the presence of adriamycin reversed the SJH-induced protection. Overall, the results suggest that SJH-associated Mn-SOD and GSH are important factors in the mechanism of the SJH-induced protective mechanism in H9C2 cardiac muscle cells. PMID- 15826872 TI - Effect of the inhibition of triiodothyronine (T3) production by thiamazole on the T3 and serotonin content of immune cells. AB - Triiodothyronine (T3) and serotonin are present in the cells of immune system (blood, peritoneal and thymic lymphocytes, monocytes and granulocytes of the blood and peritoneal fluid, mast cells). In the present experiments the effect of thiamazole, an antithyroid drug was studied on the content of these two hormones in immune cells after one and two weeks of continuous treatment (by drinking water, containing 30 mg/100 ml thiamazole, ad libitum) in adult male rats, using flow cytometric and confocal microscopic analysis. In thymic lymphocytes both hormone contents significantly increased in both time points. A significant decrease of T3 was observed in peritoneal monocytes and granulocytes also in both time points, in peritoneal mast cells after one week and in blood lymphocytes after two weeks. Serotonin content in addition to the elevated thymic values (in both measurements) was significantly reduced in blood lymphocytes after two weeks. Confocal microscopy demonstrated heterogeneous cell populations with disparate hormone content and mostly diffuse localization The experiments call attention to the presence of T3 in the immune cells and to its variable concentration under the effect of a thyrostatic drug as well, as to the T3 serotonin relationship in the cells of the immune system. PMID- 15826873 TI - Preferred conformations of endogenous cannabinoid ligand anandamide. AB - Anandamide (arachidonyl-ethanolamide, AEA) is an important endogenous cannabinoid ligand isolated from porcine brain. AEA has a flexible molecular structure with a series of four non-conjugated double bonds, a hydrophobic alkyl chain, and a carboxyamide head group. It is known that AEA binds to cannabinoid receptor and induces cannabimimetic activity. However, questions still remain about the three dimensional arrangement of the pharmacophoric groups of AEA that facilitate its interaction with cannabinoid receptor, a member of transmembrane G-protein coupled receptors (GPCRs). Such information is of critical importance for the design of novel analogs of potential therapeutic values. In the present studies, we developed a combined approach of 2D high-resolution NMR and computer modeling to investigate conformational features of AEA in solution. The developed method and experimental data is then applied to study the structural properties of AEA in a membrane-like environment that will be reported elsewhere. In addition to the measured NOEs, the dihedral angle constraints were for the first time being used as experimentally-determined structural constraints for performing molecular dynamics simulations to refine the NMR-determined AEA conformations. Our results showed that AEA prefers an extended pseudo-helical conformation in solution with two oxygen atoms pointing towards the same side and a straight pentyl chain, which was an averaged conformation observed on the basis of NMR time scale. The results were correlated to the computer predicted AEA models reported by others. The established NMR-based computational approach provides an alternative way to explore further the detailed conformational properties of AEA that encodes important pharmacophoric and conformational information regarding the activation of cannabinoid receptors. PMID- 15826874 TI - Correlation of susceptibility to 6-aminonicotinamide and hydrocortisone-induced cleft palate. AB - Our previous genome-wide Quantitative Trait Locus (QTL) mapping study using mouse A/J by C57BL/6J recombinant inbred (RI) lines suggested several chromosomal regions contain genes influencing susceptibility to phenytoin (PT)-induced cleft lip with or without cleft palate [CL(P)] and 6-aminonicotinamide (6-AN)-induced isolated cleft palate (CP). Importantly, the same chromosomal regions but different RI parental strain alleles were sometimes implicated in susceptibility to these different kinds of orofacial clefting. Here we report the susceptibility to hydrocortisone (HC)-induced CP in these RI lines. We treated pregnant females with HC and studied the incidence of CP in day 17 fetuses. RI lines showed highly correlated responses to HC and 6-AN. The A/J parental line and five RI lines showed very high levels of clefting in response to both of these teratogens. The C57BL/6J parental line and five other RI lines exhibited low incidence of CP for these teratogens. In contrast, there was no significant correlation between incidence of PT-induced CL(P) and HC-induced CP. PMID- 15826875 TI - Skin temperature rise induced by calcitonin gene-related peptide in gonadotropin releasing hormone analogue-treated female rats and alleviation by Keishi-bukuryo gan, a Japanese herbal medicine. AB - The effects of Keishi-bukuryo-gan on calcitonin gene-related peptide (CGRP) induced elevation of skin temperature were investigated in gonadotropin-releasing hormone (GnRH) analogue-treated female rats. Leupline (1.0 mg/kg) as the GnRH analogue was subcutaneously (s.c.) injected into female rats. After Keishi bukuryo-gan (100-1,000 mg/kg, p.o.) or 17beta-estradiol (0.010 mg/kg, s.c.) was administered to GnRH analogue-treated rats for 14 days, CGRP-induced skin temperature elevation, concentration of plasma 17beta-estradiol and pituitary gonadotropin (luteinizing hormone; LH, and follicle stimulating hormone; FSH) were measured. In addition, effects of 17beta-estradiol and Keishi-bukuryo-gan on the proliferation of estrogen-dependent human breast cancer (MCF-7) cells were investigated under in vitro conditions. GnRH analogue significantly lowered the concentrations of plasma 17beta-estradiol and pituitary gonadotropins. Tissue weights of the ovaries and uterus were also decreased by the analogue. Under the condition of estrogen deficiency, intravenous (i.v.) injection of exogenous CGRP (10 microg/kg) elevated the skin temperature of the hind paws more significantly than it did in sham-treated control rats. Estrogen supplementation inhibited this elevation of skin temperature with restoration of both the lowered plasma estrogen level and the decreased uterine weight in GnRH analogue-treated rats. On the other hand, Keishi-bukuryo-gan inhibited the elevation of skin temperature in a dose-dependent manner without restoring the plasma estrogen level and uterine weight. In addition, in an in vitro study, MCF-7 cells proliferated in a dose dependent manner by the addition of 17beta-estradiol (10(-13)-10(-8) M) to the medium. However, Keishi-bukuryo-gan (10(-6)-10(-4) mg/ml) did not activate the MCF-7 cell proliferation. These results suggest that Keishi-bukuryo-gan, which does not exhibit estrogen activity, may be useful for the treatment of hot flashes in women who are undergoing medical ovariectomy with a GnRH analogue. PMID- 15826876 TI - Effect of miconazole on intracellular Ca2+ levels and proliferation in human osteosarcoma cells. AB - The effect of miconazole, an anti-fungal drug, on cytoplasmic free Ca2+ concentrations ([Ca2+]i) in human osteosarcoma cells (MG63) was explored by using the Ca2+-sensitive dye fura-2. Miconazole acted in a concentration-dependent manner with an EC50 of 75 microM. The Ca2+ signal comprised a gradual rise and a sustained elevation. Removal of extracellular Ca2+ reduced 50% of the signal. In Ca2+-free medium, the [Ca2+]i rise induced by 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) was completely inhibited by pretreatment with 20 microM miconazole. Pretreatment with thapsigargin partly inhibited miconazole-induced Ca2+ release. The miconazole-induced Ca2+ release was not changed by inhibition of phospholipase C with 2 microM U73122. By using tetrazolium as a fluorescent probe, it was shown that 10-100 microM miconazole decreased cell proliferation rate in a concentration-dependent manner. Collectively, this study shows that miconazole induces [Ca2+]i rises in human osteosarcoma cells via releasing Ca2+ mainly from the endoplasmic reticulum in a manner independent of phospholipase C activity, and by causing Ca2+ influx. Furthermore, miconazole may be cytotoxic to the cells at higher concentrations. PMID- 15826877 TI - Inhibitory avoidance memory retention in the elevated T-maze is impaired after perivascular manipulation of the common carotid arteries. AB - Perivascular manipulation promoted by the positioning of a silicone collar around the common carotid arteries causes local inflammation and has been suggested as an animal model of atherosclerosis. This manipulation induces biochemical and morphological changes that are similar to those observed in the early stage of atherosclerosis in humans. Based on evidences showing that atherosclerosis is associated with cognitive deficits in humans, we presently investigated the temporal consequences of the bilateral positioning of silicone collars around the common carotid arteries (n = 15) on inhibitory avoidance memory retention in male Wistar rats tested in the elevated T-maze. The effects of this procedure were compared to those observed in sham-operated animals (n = 15) and to those observed in animals submitted to permanent bilateral occlusion of the common carotid arteries (n = 16). Additionally we studied the effects of the pretreatment with the non-selective anti-inflammatory drug indomethacin (n = 13) or the selective COX-2 inhibitor celecoxib (n = 12) and compared the effects to those of the pretreatment with vehicle (n = 11). The results showed that the silicone collar implants induced deficits in memory retention when animals were tested 2 and 4, but not 15 or 30, days after surgery. Permanent bilateral occlusion of the common carotid arteries impaired avoidance retention up to 30 days after surgery. Pretreatment with indomethacin (2 mg/kg/day) or celecoxib (5 mg/kg/day) post surgery and up to 3 days thereafter did not prevent memory deficits caused by silicone collar implants. Our data suggest that the prostanoids that participate in the inflammatory process triggered by the placement of the silicone collar do not seem responsible for the deficit in memory retention observed during the first days after collar placement. PMID- 15826878 TI - Effect of lycium barbarum polysaccharide on human hepatoma QGY7703 cells: inhibition of proliferation and induction of apoptosis. AB - Lycium barbarum polysaccharide (LBP), extracted from Lycium barbarum that is a kind of traditional Chinese herb, is found to have anticancer activity. In this study, the effect of LBP on the proliferation rate, cell cycle distribution and apoptosis in the human hepatoma QGY7703 cell line were investigated. The effects of this compound were also tested on the concentration of calcium in cells. LBP treatment caused inhibition of QGY7703 cell growth with cycle arrest in S phase and apoptosis induction. The amount of RNA in cells and the concentration of intracellular Ca2+ were increased. Moreover, the distribution of calcium in cells was changed. Taken together, the study suggests that the induction of cell cycle arrest and the increase of intracellular calcium in apoptotic system may participate in the antiproliferative activity of LBP in QGY7703 cells. PMID- 15826879 TI - Naringenin attenuates cisplatin nephrotoxicity in rats. AB - The effect of naringenin (NAR), a naturally occurring citrus flavanone, on the acute nephrotoxicity produced by cisplatin (7 mg/kg, i.v.) was investigated in the rat. Oral administration of NAR (20 mg/kg/day) for 10 days, starting 5 days before cisplatin single i.v. injection, produced significant protection of renal function. NAR reduced the extent of cisplatin-induced nephrotoxicity, as evidenced by significant reduction in serum urea and creatinine concentrations, decreased polyuria, reduction in body weight loss, marked reduction in urinary fractional sodium excretion and glutathione S-transferase (GST) activity, and increased creatinine clearance. Cisplatin-induced alterations in renal cortex lipid peroxides and GST activity were markedly improved by NAR. Cisplatin-induced alterations in renal cortex antioxidant defense system were greatly prevented by NAR. In cisplatin-NAR combined treatment group, antioxidant enzymes namely superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) were significantly increased to 54.5, 30.3 and 35.6%, respectively compared to cisplatin treated group. Platinum renal content was not affected by NAR treatment. The results provide further insight into the mechanisms of cisplatin induced nephrotoxicity and confirm the antioxidant potential of NAR. PMID- 15826880 TI - Prospective evaluation of folic acid supplementation on plasma homocysteine concentrations during weight reduction: a randomized, double-blinded, placebo controlled study in obese women. AB - Elevated total homocysteine concentrations and obesity are both associated with an increased risk of cardiovascular disease. However, previous studies of weight reduction on serum homocysteine concentrations have obtained inconsistent reports. We investigated the effect of folic acid supplementation on serum homocysteine concentrations via a randomized, double-blinded, placebo-controlled study. Seventy-four obese women [age (mean +/- SEM) 41 +/- 1 years; body mass index, 29.6 +/- 0.5 kgs/m2] completed a 12 weeks weight reduction program with dietary advice and light exercise. They were also randomized to take either folic acid supplementation (5 mg daily, n = 36) or placebo (n = 38) groups. This program led to a weight reduction of 7.7% and 8.9% of initial weight for folic acid supplementation and placebo groups, respectively. Serum folate concentrations increased for 3 folds (p < 0.001) in the folic acid group. In the folic acid group, there was a trend of lower fasting serum homocysteine concentrations (7.6 +/- 0.2 vs. 7.3 +/- 0.3 micromol/L), but it did not reach statistical significance (p = 0.170). However, we found that serum homocysteine concentrations decreased significantly in those with higher baseline homocysteine concentrations (8.7 +/- 1.3 vs. 7.8 +/- 1.5 micromol/L, p = 0.004), while it did not change in those with lower baseline homocysteine concentrations (6.6 +/- 0.6 vs. 6.8 +/- 1.2 micromol/L, p = 0.334). Reduction of serum homocysteine concentrations did not correlate with elevation of serum folate concentrations (p = 0.646) in obese women with higher baseline homocysteine concentrations. In conclusion, serum homocysteine concentrations can be maintained in obese women during mild to moderate weight loss. Folic acid supplementation decreased serum homocysteine concentrations in those women who had higher serum homocysteine concentrations before participating in the weight reduction program. PMID- 15826885 TI - Counting counts. PMID- 15826886 TI - Measuring the burden of premature mortality in Pakistan: use of sentinel surveillance systems. AB - INTRODUCTION: In developing countries, finding valid and reliable data is a great challenge. The Federal Bureau of Statistics in Pakistan maintains a surveillance system that records vital events. Results are reported as the Pakistan Demographic Survey (PDS). METHODS: The adjusted mortality data from PDS-1997 was used to estimate the burden of premature mortality using the healthy life year (HeaLY) methodology. RESULTS: The burden of premature mortality for Pakistan was 367 HeaLYs lost per 1000 population. More than half of the HeaLYs are being lost due to infection, maternal and perinatal conditions, and malnutrition. CONCLUSION: In the absence of a system maintaining reliable information regarding vital events, PDS provides a good alternate source of such information that can be used to aid policy making for health care and future research. PMID- 15826887 TI - Unintentional injuries over a 1-year period in a rural Vietnamese community: describing an iceberg. AB - OBJECTIVES: To document unintentional injuries in a rural community over a 1-year period as a basis for prioritizing preventive activities. STUDY DESIGN: Quarterly home visits over 1 year to elicit experience of injury among household members in the preceding 3 months. METHODS: In total, 24,776 people living in rural communities in the Bavi District, Northern Vietnam, were surveyed in home visits during 2000. In the home visits, injuries that needed care or disrupted normal activities were recorded, together with their circumstances. RESULTS: Overall, 2079 new non-fatal injuries were recorded over 23,338 person-years, a rate of 89/1000 person-years-at-risk. Males had a significantly higher injury rate than females for all age groups except for those aged 35-59 years and the elderly (P<0.05). The elderly were at highest risk of injury (P<0.05), particularly females. Home injuries occurred at the highest overall rate, particularly among the elderly. Road traffic injuries were most common among children. Most injuries involved contact with another object. Less than one-quarter of injury victims sought care at a health facility. CONCLUSIONS: Community-based household surveys revealed the hidden part of the injury iceberg, as well as showing high incidence rates, indicating that injury is an important public health problem which should be a priority for intervention in rural Vietnam, and probably elsewhere. This comprehensive study is intended to contribute evidence and methods to the Ministry of Health's national programme for injury prevention, and to a wider audience. PMID- 15826888 TI - Applying a basic development needs approach for sustainable and integrated community development in less-developed areas: report of ongoing Iranian experience. AB - INTRODUCTION: Despite considerable achievements in the provision of basic developmental facilities in terms of drinking water, access to primary healthcare services, high-quality and nutritious food, social services, and proper housing facilities, there are many rural and slum communities in Iran where these essential needs remain unfulfilled. Lack of equity is prominent, as large differences exist in underprivileged provinces. New policies developed in the past two decades have resulted in substantial achievements in meeting population needs and reducing the socio-economic gap; nevertheless, poverty levels, unemployment due to a large increase in the birth rate in the early 1980s, and lack of community participation are matters yet to be addressed. To overcome these deficiencies, a basic development needs approach was adopted to promote the concept of community self-help and self-reliance through intersectoral collaboration, creating an environment where people could take an active part in the development process, with the Iranian government providing the necessary support to achieve the desired level of development. DESCRIPTION OF THE PROJECT: Following firm commitment from the Iranian government and technical support from the World Health Organization Regional Office, basic development needs was assigned a high priority in health and health-related sectors, reflected in the third National Masterplan (2001-2005). A comprehensive intersectoral plan was designed, and pilot projects were commenced in three villages. Each village elected a representative, and committee clusters were formed to run and monitor projects identified by a process of local needs assessment and priority assignment. In each region, a variety of needs were elicited from these assessments, which were actively supported by local authorities. LESSON LEARNED: A basic development needs approach was found to be a reliable discipline to improve community participation, needs-led resource allocation and intersectoral co-operation in community development, particularly in underprivileged areas. Iran's initial experience of basic development needs has gained widespread public support but will require periodical evaluation as it is introduced into other rural and urban regions across the country. PMID- 15826889 TI - Epidemiology of oesophageal atresia and tracheo-oesophageal fistula in Hawaii, 1986-2000. AB - The most common major birth defects affecting the oesophagus and trachea are oesophageal atresia (OA) and tracheo-oesophageal fistula (TOF). Epidemiological information on OA and/or TOF in the USA has been derived from only a few populations. Sixty-one cases of OA or TOF were identified through a population based birth defects registry in Hawaii during 1986-2000. The relationship between various clinical and demographic factors and OA or TOF were examined. The rate of OA or TOF was 2.2 per 10,000 live births. The cases were 67.2% OA with TOF, 21.3% TOF alone, and 11.5% OA alone. Risk of OA or TOF was significantly higher among multiple births and live births with low birth weight and gestational age. The risk of OA or TOF was significantly higher for whites. The rate of OA or TOF was consistent with that reported elsewhere. The risk of the defects differed significantly between the major racial/ethnic groups in Hawaii. The impact of many factors on OA or TOF in Hawaii was similar to that reported elsewhere. PMID- 15826890 TI - Prevalence and risk factors associated with dyslipidaemia in children and adolescents among ethnic groups in Taiwan. AB - BACKGROUND: The mortality rates of cerebral and cardiovascular diseases are higher for aborigines than non-aborigines in Taiwan. Hypertriglyceridaemia and hypercholestolaemia are risk factors for cardiovascular diseases. OBJECTIVES: To investigate the prevalence of dyslipidaemia and its associated risk factors in aborigine (Atayal, Paiwan and Bunun tribes) and non-aborigine (Fukein and Hakka Chinese) children and adolescents in Taiwan. STUDY DESIGN: This was a cross sectional study. METHODS: In total, 718 males and 721 females, below 20 years of age, were recruited. Our study defined dyslipidaemia as serum triglyceride and cholesterol levels greater than 200 and 240 mg/dl, respectively. RESULTS: The serum triglyceride level and the prevalence of hypertriglyceridaemia were similar in both aborigines and non-aborigines and both sexes, but the Bunun and Paiwan tribes had the highest prevalence of hypertriglyceridaemia in males (11.8-29.4%) and females (10.9-22.8%) compared with other aboriginal tribes (5.1-10.8% for males and 7.8-9.2% for females). Serum cholesterol concentrations and the prevalence of hypercholesterolaemia were lower in the aborigines than non aborigines for both sexes (P<0.05), with the Atayal tribe having the lowest prevalence in males (1.1%) and females (2.1%) compared with other aboriginal tribes (2.4-4.5% for males and 5.7-8.0% for females). Using multivariate-adjusted logistic regression modelling, hypertriglyceridaemia was significantly associated with the Bunun tribe (odds ratio (OR)=3.2, 95% confidence intervals (CI) 1.6 6.1), hyperuricaemia (OR=1.8, 95% CI 1.2-2.6), hypercholesterolaemia (OR=3.3, 95% CI 1.7-6.4) and alcohol use (OR=2.8, 95% CI 1.2-6.6). Hypercholesterolaemia, after controlling for age and sex, was significantly associated with the Atayal tribe (OR=0.2, 95% CI 0.1-0.5), hypertriglyceridaemia (OR=3.5, 95% CI 1.8-6.7) and hyperuricaemia (OR=3.2, 95% CI=1.7-6.0). CONCLUSIONS: For the young people of Taiwan, hypertriglyceridaemia is associated with hyperuricaemia, hypercholesterolaemia and alcohol use, and hypercholesterolaemia is associated with hypertriglyceridaemia and hyperuricaemia. Compared with non-aborigines, the young aborigines of some tribes have a higher prevalence of hypertriglyceridaemia and a lower serum cholesterol level. PMID- 15826891 TI - How education, income, control over life and life style contribute to risk factors for cardiovascular disease among adults in a post-communist country. AB - OBJECTIVES: To examine the effect of education, income, food habits and control over life on risk factors for cardiovascular diseases in Poland, a former communist country with ongoing transformations of the political and economic system, where cardiovascular disease is an important health problem. METHODS: The data were collected by questionnaire, as part of the CINDI Programme, from 2001 to 2002 in Lodz, an industrial city in Poland. The surveys were directed at random samples of 2000 men and 2000 women; 1847 participants aged 18-64 years responded. As detailed in the ATP III report, participants with three or more of abdominal obesity, hypertriglyceridaemia, low high-density-lipoprotein cholesterol, high blood pressure or high fasting glucose were defined as having the metabolic syndrome. RESULTS: We have found that education is more strongly associated with the metabolic syndrome. Our study suggests that higher education may be a strong and consistent predictor of good health. Also, low control over life (unfulfilled daily needs, unfulfilling work, little or no reward for effort at work) and lack of physical activity have been shown to be predictors for the metabolic syndrome. CONCLUSIONS: Our findings suggest that interventions to improve the health of the Polish population need to be specific for the social environment. PMID- 15826892 TI - Poor health is associated with episodic heavy alcohol use: evidence from a National Survey. AB - OBJECTIVES: The objective of this study was to examine the relationship between self-rated health and episodic heavy drinking in a representative sample of American adults. We also sought to determine ethnic and gender differences in the association between self-rated health and episodic heavy drinking. METHODS: Data (n=4649) from the Third US National Health and Nutrition Examination Survey were utilized for this investigation. Episodic heavy drinking was defined as the consumption of five or more and four or more alcoholic beverages on one occasion for men and women, respectively. Poor health was defined as answering fair or poor to the question: "Would you say your health in general is excellent, very good, good, fair or poor?" Odds ratio from the logistic linear regression analysis was used to estimate the risk for poor health that was associated with episodic heavy drinking. Statistical adjustments were made for age, hypertension, diabetes, current smoking, body mass index and race/ethnicity. RESULTS: Overall, episodic heavy drinking was associated with increased odds of poor self-rated health in men and women. In men, episodic heavy drinking was independently associated with 1.28 (95% CI: 1.07-1.82) increased odds of poor health. The corresponding value in women was 1.86 (95% CI: 1.05-2.28). In men, being Black was associated with approximately two-fold (OR=1.96; 95% CI: 1.33, 2.89), and being Hispanic was associated with approximately four-fold (OR=3.59; 95% CI: 2.50, 5.14) increased odds of poor self-rated health relative to being White. The corresponding odds ratios in women were 2.97 (95% CI: 1.90, 4.64) and 5.18 (95% CI: 3.23, 8.30). Associations were greater among blacks (adjusted OR=2.41; 95% CI: 1.81-3.22) and Hispanics (adjusted OR=4.15; 95% CI: 3.12-5.52) than among whites. CONCLUSIONS: Poor health is associated with episodic heavy alcohol consumption. Public health strategies to curb alcohol abuse may improve self reported health status in these at-risk populations. PMID- 15826893 TI - Sedentary behaviour, physical activity and weight problems in adolescents in Wales. AB - OBJECTIVE: We studied the prevalence and stability of overweight and obesity in a cohort of adolescents, and the effects of sedentary behaviour and physical activity on changes in body mass. The study also examined the extent to which physical activity mediated the relationship between sedentary behaviour and body mass. DESIGN: Four-year cohort study. METHODS: The study was part of the Health Behaviour of School-aged Children Study that took place in Wales between 1994 and 1998. Body height and weight measurements and self-report data on sedentary behaviour, physical activity and psychosocial adjustment were collected from 355 adolescents on two occasions 4 years apart. The mean age of the sample at baseline was 12.30 (SD=6.30) years. RESULTS: Weight conditions (underweight, overweight and obesity) and body mass were moderately stable over the interval. Regression analyses showed that sedentary behaviour at Time 1 predicted body mass at Time 2, while physical activity predicted a change in body mass over time. The influence of sedentary behaviour on body mass was not found to be mediated by physical activity. However, weight problems in Year 7 coincided with getting bullied, bullying others, and feeling left out of things. Obesity was also related to snacking and skipping breakfast. CONCLUSIONS: Sedentary behaviour and physical activity in early adolescence both influenced body mass in late adolescence. Results indicated that promoting healthy diets and physical activities may have long-term health benefits for young people. PMID- 15826894 TI - A review of UK housing policy: ideology and public health. AB - OBJECTIVES: The aim of this paper is to review UK public health policy, with a specific reference to housing as a key health determinant, since its inception in the Victorian era to contemporary times. REVIEW: This paper reviews the role of social and private housing policy in the development of the UK public health movement, tracing its initial medical routes through to the current socio economic model of public health. The paper establishes five distinct ideologically and philosophically driven eras, placing public health and housing within liberal (Victorian era), state interventionist (post World War 1; post World War 2), neoliberal (post 1979) and "Third Way" (post 1997) models, showing the political perspective of policy interventions and overviewing their impact on public health. The paper particularly focuses on the contemporary model of public health since the Acheson Report, and how its recommendations have found their way into policy, also the impact on housing practice. CONCLUSIONS: Public health is closely related to political ideology, whether driven by the State, individual or partnership arrangements. The current political system, the Third Way, seeks to promote a sustainable "social contract" between citizens and the State, public, private and voluntary organizations in delivering community-based change in areas where health inequalities can be most progressively and successfully addressed. PMID- 15826895 TI - Effect of indoor air quality in the postnatal period on lung function in pre adolescent children: a retrospective cohort study in Poland. AB - The purpose of this study was to determine the association between level of lung function in pre-adolescence and indoor air quality in the postnatal period. The retrospective cohort study was carried out in a sample of 1036 pre-adolescent children (9 years of age) attending schools in two residential areas of Krakow, Poland. Measurement of health outcomes considered lung function together with height and weight. Indoor air quality was based on environmental tobacco smoke and type of household heating. In addition, the number of winter months that occurred during the first 6 months of life was included as a key independent variable. Multivariate linear regression of lung function measured by forced expiratory volume in 1s (FEV1) and forced vital capacity (FVC), and postnatal exposure to indoor pollution in the heating season (PEIP score) was adjusted for potential confounders such as maternal smoking during pregnancy and parental education as a proxy of social class. In the total study sample, the adjusted beta coefficient for FEV1 per unit of the PEIP score was -0.06 (P=0.02), while that for FVC was -0.05 (P=0.04). The analysis carried out in the more polluted area found that children living in households heated with gas or coal had a PEIP score that was strongly inversely related to lung function (adjusted beta coefficient for FEV1=-0.13; P=0.03; for FVC=-0.15, P=0.01), whereas regression coefficients were not significant in the group of children living in households with central heating. This study suggests that a lower level of lung function in pre-adolescent children can be related to postnatal exposure to indoor emissions in the winter. PMID- 15826896 TI - Polio immunization policy in Mexico: economic assessment of current practice and future alternatives. AB - The World Health Organization recommends that all children aged less than 5 years should be vaccinated against polio through intensive immunization programmes as well as routine immunization. A national immunization week (NIW) was held in February 2002 in the Monterrey district of Mexico. A prospective micro-costing study was conducted to measure the total cost to the state of the NIW, the cost profile, and the ratio of cost per immunization contact to cost per dose of oral polio vaccine (OPV), and to compare OPV and inactive polio vaccine (IPV) in economic terms. Two scenarios were used as the basis for calculation. The cost of volunteers was excluded from the "lower-cost scenario" and included in the "upper cost scenario". The total cost of the NIW was USD 100,454 for the lower-cost scenario and USD 156,614 for the upper-cost scenario. The major part of the costs was personnel costs (67.30 and 77.53% of the total costs in the lower- and upper cost scenario, respectively). The ratio of cost per immunization contact to cost per dose of OPV was 6.45 for the lower-cost scenario and 10.05 for the upper-cost scenario. Changing from the current OPV-based intensive and routine schedule to a sequential IPV-OPV routine schedule would save USD 14.52 per vaccinated child, and changing to a full IPV routine schedule would save USD 9.41 per vaccinated child. PMID- 15826897 TI - Seroprevalence of pertussis among children in Eastern Turkey. AB - The aim of this cross-sectional study was to determine the immunity status of children to pertussis by socio-demographic characteristics in Eastern Turkey. The study sample consisted of 840 randomly selected and healthy children aged 0-71 months. The seroprevalence of pertussis was 30.1%. Age, parent education and economic status were not associated with the geometric mean titers (GMT) of pertussis antibody, while gender, residential area and the application number of diphtheria-tetanus-whole-cell pertussis vaccine were associated with GMT. Most preschool children are susceptible to pertussis and current vaccination efforts do not provide adequate immunization. PMID- 15826898 TI - Dental caries in preschool children in Bilac, Brazil. PMID- 15826907 TI - Molecular characterization of tomato-infecting begomoviruses in Thailand. AB - Bipartite geminiviruses infecting tomatoes in Thailand were detected by polymerase chain reaction (PCR) using CPA5/CPA2 primers. Products derived from PCR-amplified full-length DNA-A and DNA-B of TYLCV collected from Chiang Mai, Nong Khai, and Sakon Nakhon were cloned and sequenced. DNA-A from Chiang Mai was 2747 nts long; Nong Khai, 2744 nts; and Sakon Nakhon, 2747 nts, and those of DNA B from Chiang Mai were 2750 nts long; Nong Khai, 2749 nts; and Sakon Nakhon, 2749 nts. The genomes of these virus isolates were organized like those of other begomoviruses. The DNA-A had two ORFs in the virion sense and four ORFs in the complementary sense. The DNA-B had two ORFs in the virion sense and one ORF in the complementary sense. Nucleotide sequences of DNA-A of TYLCV from Chiang Mai, Nong Khai, and Sakon Nakhon were closely related to those of Tomato yellow leaf curl Thailand virus (TYLCTHV) and Tomato yellow leaf curl Thailand virus [Myanmar] (TYLCTHV-[MM]) with nucleotide sequence identity ranging from 89% to 95%. Based on sequence comparisons and phylogenetic analyses, these three virus isolates studied were identified as new strains of TYLCTHV and named Tomato yellow leaf curl Thailand virus-Chiang Mai (TYLCTHV-[CM]The GenBank accession codes for DNA-A of TYLCTHV-[CM], -[NK], and -[SK] are , and , respectively. The GenBank accession codes for DNA-B of TYLCTHV-[CM], -[NK], and -[SK] are , , and , respectively.), Nong Khai (TYLCTHV-[NK] and Sakon Nakhon (TYLCTHV-[SK]). PMID- 15826908 TI - Expression and analysis of the Epstein-Barr virus BARF1-encoded protein from a tetracycline-regulatable adenovirus system. AB - Epstein-Barr virus (EBV) has been associated with human cancers of lymphocytic or epithelial origin. Potential functions of the BARF1 early gene in EBV oncogenesis emerged from our observations showing expression of BARF1-encoded protein in nasopharyngeal carcinoma biopsies, and induction of either malignant transformation (in rodent fibroblast and human B cell lines) or immortalization (in monkey primary epithelial cells) following BARF1 transfection. We previously reported expression of the BARF1 product as a cytoplasm/membrane-associated protein from 293-tTA cells infected with a BARF1-recombinant adenovirus. Since constitutive expression of BARF1 from this heterologous system became inefficient, we developed a tetracycline-regulatable recombinant vector expressing BARF1 and green fluorescent protein from a dicistronic message. As here reported, stable and efficient expression of BARF1 from this vector in either permissive or non-permissive cell lines, allowed the first sequencing identification and further molecular characterization of BARF1-encoded protein. PMID- 15826909 TI - Cotton leaf curl Gezira virus-satellite DNAs represent a divergent, geographically isolated Nile Basin lineage: predictive identification of a satDNA REP-binding motif. AB - Cotton leaf curl Gezira virus (CLCuGV), a species of the genus Begomovirus (family Geminiviridae), was recently cloned from cotton, okra, and Sida alba plants exhibiting leaf-curling and vein-thickening symptoms in Sudan. Here, we describe a previously unknown lineage of single-stranded DNA satellite (satDNA) molecules, which are associated with CLCuGV, and are required for development of characteristic disease symptoms. Co-inoculation of cotton and Nicotiana benthamiana plants with satDNAs cloned from cotton, okra, and S. alba, together with CLCuGV as the 'helper virus' resulted in the development of characteristic leaf-curling and vein-thickening symptoms in both hosts. An anatomical study of symptomatic, virus-infected cotton leaves revealed that spongy parenchyma cells had developed instead of collenchyma cells at the sites of vein thickening. Phylogenetically, the CLCuGV-associated satDNAs from Sudan, together with their closest relatives from Egypt, form a new satDNA lineage comprising only satDNAs from the Upper and Lower Nile Basins. Analysis of satellites and their helper virus sequences identified a predicted REP-binding site consisting of the directly repeated sequence, 'CGGTACTCA', and an inverted repeated sequence, 'TGAGTACCG', which occur in the context of a 17-nucleotide motif. The conserved REP-binding motif identified herein, together with strict geographic isolation, and apparent host-restriction, may be the collective hallmark of these new satDNA begomovirus lineages, extant in the Nile Basin. PMID- 15826910 TI - Composition bias and genome polarity of RNA viruses. AB - I have observed a relationship between GC content in coding sequences of RNA viruses and their genome polarity. Positive-stranded RNA viruses have significantly higher GC contents than negative-stranded RNA viruses. Coding sequences of all negative-stranded RNA viruses are biased toward high A in coding strands (high T in genomes), while two distinct patterns were observed among positive-stranded RNA genomes. This finding suggests that RNA viruses with different genome polarity are under different mutational pressure, which may be a consequence of the difference in the strategies of viral genome expression and replication. The GC content directly affects the viral codon adaptation index using highly expressed human genes as the reference set, which may theoretically predict the efficiency of viral gene expression in human cells. PMID- 15826911 TI - Avipoxvirus multiplication in a mammalian cell line. AB - Avipoxviruses have many advantages and are being increasingly employed as recombinant vaccine vectors. One attractive feature is that while inserted transgenes are expressed in immunologically favourable ways, avipoxvirus infections of mammalian cells are believed to be abortive. The experimental evidence supporting this belief is, however, based on a limited number of mammalian cell-types and a few avipoxvirus species. We evaluated two avian and eight mammalian cell lines for permissivity to three avipoxvirus strains, one reference fowlpoxvirus and two newly isolated strains from sparrow and pigeon, respectively. Both avian cell lines were, as expected, permissive for all three avipoxvirus strains. However, by multiplication assays, we found to our surprise that Syrian baby hamster kidney (BHK-21) cells were equally permissive to all virus strains. Results from electron microscopy of infected BHK-21 cells revealed viral morphogenesis proceeding to various forms of infectious viruses. These results were supported by the demonstration of avipoxvirus specific late gene expression and avipoxvirus specific DNA restriction pattern in BHK-21 infected cells. PMID- 15826912 TI - Promoter activity of left inverted terminal repeat and downstream sequences of porcine adenovirus type 3. AB - Early region 1 (E1) of porcine adenovirus type 3 (PAdV-3) consists of E1A and E1B transcription units. The authentic promoter region of E1A contains a TATA box at nucleotide position (nt) 449 and a bifunctional regulatory element between nt 374 and 431, which enhances the transcription of E1A, but represses that of E1B. Here, we investigated the role of the left inverted terminal repeat (ITR) and its downstream sequences (between nt 151 and 312) in the transcription of early viral genes, and viral replication. Mutant PAdV-3s without the authentic E1A promoter region could be rescued by transfection of mutant genomic DNA into fetal porcine retina cells. Moreover, the mutant PAdV-3s produced E1A-specific mRNA and remained viable in swine testis (ST) cells suggesting that the left-terminal 151 bp including the ITR, can serve as a promoter for E1A expression. However, mutant PAdV-3s containing deletion including authentic E1A promoter region, displayed both reduced steady-state levels of early gene mRNAs (E1A, E1B, E2A, E3, and E4) and decreased rate of viral replication in ST cells. Interestingly, mutant PAdV 3s containing the left-terminal 312 bp displayed increased transcription of early genes including E1A. Our results suggest that the left ITR of PAdV-3 contain the promoter like elements and the sequences (between nt 151 and 312) downstream of left ITR can enhance its promoter activity. PMID- 15826913 TI - Phylogenetic and restriction fragment length polymorphism analyses of hemagglutinin (H) protein of canine distemper virus isolates from domestic dogs in Japan. AB - We conducted phylogenetic and restriction fragment length polymorphism (RFLP) analyses of 995 nucleotides within the hemagglutinin (H) gene open reading frame (ORF) of field isolates of 23 canine distemper virus (CDV) strains isolated from domestic dogs in Japan between 1982 and 1998. The phylogenetic analysis showed that Japanese field isolates could be separated into three groups. Eighteen out of the twenty-three strains constituted one cluster consisting of Japanese CDVs, four strains formed a second Japanese CDV group, and only one strain belonged to a group containing foreign CDV strains. By RFLP analysis using SspI, we could distinguish all the Japanese field isolates from the vaccine strains. Thus, the RFLP method is useful for differentiating the infections with field CDV strains from the vaccine strains in clinical cases. PMID- 15826914 TI - Early activation of the mitochondrial apoptotic pathway in Vesicular Stomatitis virus-infected cells. AB - Vesicular Stomatitis Virus (VSV) has been shown to induce apoptosis in a caspase dependent manner, but the precise apoptotic pathway remains unknown. We found that caspases 9 and 3, but not caspase 8, were activated during VSV-induced apoptosis in infected Vero cells. Since caspase 9 is related to the mitochondrial apoptotic pathway, we analyzed some mitochondrial events such as changes in the mitochondrial transmembrane potential (Deltapsim) and mitochondrial release of apoptogenic proteins such as cytochrome c and the apoptosis inducing factor (AIF). We found that VSV infection triggers the dissipation of the Deltapsim and the release of both cytochrome c and AIF from the mitochondrial intermembrane space very early in the VSV infection. These results indicate that the trigger of apoptosis in VSV-infected cells occurs through the early activation of the mitochondrial apoptotic pathway. On the other hand, intracellular levels of the anti-apoptotic proteins, such as Bcl-2 and Bcl-xL, and the pro-apoptotic protein Bax, were assessed during viral infection. These analyses showed that as viral infection proceeded, the cellular level of Bcl-xL decreased, while the levels of Bax and Bcl-2 remained unaffected. The significance of the Bcl-xL modulation is also discussed. PMID- 15826915 TI - Co-infection by two distinct totivirus-like double-stranded RNA elements in Chalara elegans (Thielaviopsis basicola). AB - A full-length cDNA clone was developed from a 5.3 kb double-stranded (ds) RNA element present in strain CKP of the plant pathogenic fungus Chalara elegans. The complete nucleotide sequence was 5310 bp in length and sequence analysis revealed that it contained three large putative open reading frames (ORFs). ORF1 was initiated at nucleotide position 329 and encoded a putative coat protein, which shared some homology (35-45% amino acid identity) to other dsRNAs in the family Totiviridae. Both ORF2 and ORF3 were initiated at nucleotide positions 2619 and 4071, respectively, and encoded a putative RNA-dependent RNA polymerase (RdRp). Sequence comparison using deduced amino acid sequences of both ORF2 and ORF3 revealed that all RdRp conserved motifs shared highest homology (41% identity) to that of SsRNA1 of Totiviridae. This dsRNA in C. elegans was designated Chalara elegans RNA Virus 1 (CeRV1). During the development of the full-length cDNA clone of CeRV1, several partial cDNA clones from an additional dsRNA fragment in strain CKP were obtained, which when aligned with each other, produced one linear fragment which was 2336 bp long. Northern blot and sequence analysis of this second clone showed it differed in sequence composition from CeRV1. This dsRNA in C. elegans was designated Chalara elegans RNA Virus 2 (CeRV2). Sequence analysis of CeRV2 showed it contained all conserved motifs and shared some homology (45% amino acid identity) to RdRp regions of Totiviridae. The nucleotide and amino acid sequences of the conserved motifs of the RdRp regions between CeRV1 and CeRV2 showed an identity of 56% and 50%, respectively. These findings suggest that co-infection of two distinct totivirus-like dsRNAs (CeRV1 and CeRV2) in C. elegans, a first report in this fungus. Transmission electron microscopy of strain CKP of C. elegans revealed the presence of putative virus-like particles in the cytoplasm, which were similar both in shape and size to viruses in the Totiviridae. PMID- 15826916 TI - Monoclonal antibodies against regions topologically surrounding the homodimeric beta-barrel interface of Epstein-Barr virus nuclear antigen-1. AB - Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is essential for maintenance of EBV latency. Four mouse monoclonal antibodies (mAbs) against the part of the EBNA-1 sequence (amino acids 451-641) containing the domain that forms a homodimeric eight-stranded beta-barrel were generated and characterized, examined for immunocytochemical staining, immunoblotting and isoelectric focusing of EBNA 1 proteins, and used to examine interactions between EBNA-1 polypeptides by far Western blot assays. Far-Western blot analyses using the mAbs suggest that both the beta-strand (aa 593-604) and alpha helix (aa 568-582) are essential for EBNA 1 dimerization, consistent with yeast two-hybrid studies of mutant EBNA-1 polypeptides. These mAbs should be useful for studies on the structure and function of EBNA-1 proteins. PMID- 15826917 TI - Genetic characterization of L-Zagreb mumps vaccine strain. AB - Eleven mumps vaccine strains, all containing live attenuated virus, have been used throughout the world. Although L-Zagreb mumps vaccine has been licensed since 1972, only its partial nucleotide sequence was previously determined (accession numbers , and ). Therefore, we sequenced the entire genome of L-Zagreb vaccine strain (Institute of Immunology Inc., Zagreb, Croatia). In order to investigate the genetic stability of the vaccine, sequences of both L-Zagreb master seed and currently produced vaccine batch were determined and no difference between them was observed. A phylogenetic analysis based on SH gene sequence has shown that L-Zagreb strain does not belong to any of established mumps genotypes and that it is most similar to old, laboratory preserved European strains (1950s-1970s). L-Zagreb nucleotide and deduced protein sequences were compared with other mumps virus sequences obtained from the GenBank. Emphasis was put on functionally important protein regions and known antigenic epitopes. The extensive comparisons of nucleotide and deduced protein sequences between L Zagreb vaccine strain and other previously determined mumps virus sequences have shown that while the functional regions of HN, V, and L proteins are well conserved among various mumps strains, there can be a substantial amino acid difference in antigenic epitopes of all proteins and in functional regions of F protein. No molecular pattern was identified that can be used as a distinction marker between virulent and attenuated strains. PMID- 15826919 TI - Thyroid peroxidase autoantibodies in euthyroid subjects. AB - Thyroid peroxidase (TPO) is a key enzyme in the formation of thyroid hormones and a major autoantigen in autoimmune thyroid diseases. Titers of TPO antibodies also correlate with the degree of lymphocytic infiltration in euthyroid subjects, and they are frequently present in euthyroid subjects (prevalence 12-26%). Even within the normal range for thyrotropin (TSH), TPO antibody titers correlate with TSH levels, suggesting that their presence heralds impending thyroid failure. Assays for serum TPO antibodies have become much more sensitive, and very low titers can be found in virtually all subjects. However, titers above an assay dependent cut-off are a clear risk factor for hypothyroidism; in the Whickham survey the annual risk of developing hypothyroidism in TPO-positive women with normal thyrotropin levels was 2.1%. Measuring TPO antibodies in euthyroid subjects can be used to identify subjects with increased risk for hypothyroidism: e.g. as triage to measure thyrotropin. This could be done in women who wish to become pregnant and those with an increased risk per se who are pregnant (to predict first trimester hypothyroidism, and postpartum thyroid dysfunction), patients with other autoimmune diseases, subjects on amiodarone, lithium, or interferon-alpha, and in relatives of patients with autoimmune thyroid diseases. PMID- 15826920 TI - Non-thyroid autoantibodies in autoimmune thyroid disease. AB - Autoimmune thyroid disease is frequently accompanied by other organ-specific and non-organ-specific diseases, most likely because there is sharing of genetic and possibly environmental susceptibility factors. These associations are well recognized in the autoimmune polyglandular syndromes; autoimmune thyroid disease is one of the three major endocrinopathies in the type 2 syndrome and occurs in around 4% of type 1 patients. This review considers the frequency of disease specific autoantibodies in patients with thyroid autoimmunity and briefly examines the role of such antibodies in performing screening for the associated conditions. Recommendations are made for using such autoantibody tests in the setting of patients with autoimmune thyroid disorders, and also for the utility of screening for thyroid autoimmunity in patients with pernicious anaemia, Addison's disease, coeliac disease, primary biliary cirrhosis, myasthenia gravis, lymphocytic hypophysitis, systemic lupus erythematosus and rheumatoid arthritis. At present, however, there are no large-scale trials that have shown the cost benefit ratio of autoantibody screening for autoimmunity screening, and clinicians must use individual judgement combined with heightened awareness to identify who to test. PMID- 15826921 TI - Thyrotropin receptor antibodies: new insights into their actions and clinical relevance. AB - The thyrotropin receptor (TSHR) is a G-protein-coupled receptor with a large ectodomain. TSH, acting via TSHR, regulates thyroid growth and thyroid hormone production and secretion. The TSHR undergoes complex post-translational processing involving dimerization, intramolecular cleavage, and shedding of its ectodomain, and each of these processes may influence the antigenicity of the TSHR. The TSHR is also the major autoantigen in Graves' disease, as well as a leading candidate autoantigen in both Graves' ophthalmopathy and pretibial myxedema. The naturally conformed TSHR is most effectively presented as an autoantigen to the immune system, causing the production of stimulating TSHR-Abs. There are also autoantibodies which block the TSHR from TSH action, and neutral TSHR-Abs which have no influence on TSH action. TSHR-Abs can be detected by competition assays of TSHR-Abs for labeled TSH, or monoclonal TSHR-Ab binding to solubilized TSHRs, or by bioassays using thyroid cells or mammalian cells expressing recombinant TSHRs. PMID- 15826922 TI - Hashimoto's encephalopathy: myth or reality? An endocrinologist's perspective. AB - Since the first description of a case of episodic encephalopathy associated with Hashimoto's thyroiditis in 1966, many cases of corticosteroid-responsive encephalopathy associated with positive antithyroid antibodies, clinical Hashimoto's thyroiditis, or spontaneous autoimmune thyroid failure have been reported. These patients have neurologic manifestations of encephalopathy unrelated to other known causes. The condition has thus been termed 'Hashimoto's encephalopathy'. The literature shows no proven association between thyroid disease and the neurologic process. Although the association of a common endocrinologic condition and a rare neurologic disease may occur by chance, this type of encephalopathy probably has an autoimmune nature and thus is more likely to occur in the background of another autoimmune condition such as autoimmune thyroid disease. Until the pathogenesis of these coincident conditions is better defined, the term 'corticosteroid-responsive encephalopathy associated with autoimmune thyroiditis' is more accurate and descriptive than Hashimoto's encephalopathy. Advances in the field may clarify this seemingly inconsistent terminology. PMID- 15826923 TI - Pituitary antibodies and lymphocytic hypophysitis. AB - Lymphocytic hypophysitis (LYH) is a pituitary disease which can cause headache, changes in visual field and pituitary dysfunction. The clinical, histopathological and morphological findings and its association with other autoimmune disorders allow LYH to be included among the autoimmune diseases. Pituitary trans-sphenoidal biopsy is thought to be the diagnostic gold standard for LYH, even if some morphological findings on hypothalamic-pituitary magnetic resonance imaging (MRI) can suggest the occurrence of this disease. Despite the fact that organ-specific antibodies are good markers of many autoimmune endocrine diseases, the pathogenetic and diagnostic roles of anti-pituitary antibodies (APAs) in LYH are still under discussion. In fact, several methods have been used to detect APAs, but the conflicting results from different methods have impaired the clinical relevance of these antibodies. Recently, APAs have been detected by an immunofluorescence method in patients with selective idiopathic hypopituitarism (particularly in those with growth-hormone deficiency) and in adults with autoimmune endocrine diseases. The results suggest that only when they are present at high titres may they be considered a good marker of pituitary involvement, and in particular of growth-hormone-producing cells. PMID- 15826924 TI - Adrenal cortex autoantibodies in subjects with normal adrenal function. AB - The recent advances in our understanding of immunology have greatly improved our knowledge about the natural history of autoimmune diseases and, in particular, of autoimmune Addison's disease (Autoimmune AD). Autoimmune AD is a chronic disorder with a long preclinical period marked by the presence of adrenal cortex autoantibodies (ACAs). In this chapter the main data on this will be analyzed. The populations with the highest risk of Autoimmune AD are first relatives of patients with AAD and patients with autoimmune diseases, particularly those with chronic hypoparathyroidism or with premature ovarian failure. The best markers to identify the subjects at risk are ACAs detected by the immunofluorescence test on human or animal tissues, or 21-hydroxylase autoantibodies (21-OHAbs) detected by radioimmunoassay (RIA). The evaluation of adrenal cortex function in these individuals includes the basal determination of adrenocorticotropic hormone (ACTH), cortisol, aldosterone, plasma renin activity and cortisol after intravenous stimulation with synthetic ACTH. The multivariate analysis of the main factors (genetics, age, gender, titers of antibodies, pre-existing disease, status of the adrenal function) revealed that the risk of future AAD depends only on the presence of high antibody titers, chronic hypoparathyroidism or chronic candidiasis and adrenal dysfunction. On the basis of these parameters the risk of future Autoimmune AD can be calculated with an equation model. Patients with different risk scores need to be monitored at different time intervals, and those at high risk need to be strictly monitored and are the ideal subjects for future prevention trials. PMID- 15826925 TI - Diabetes-related antibodies in euglycemic subjects. AB - Type 1 diabetes mellitus is preceded by autoimmunity against the insulin producing islet beta cells. Autoantibodies against islet antigens such as insulin, glutamic acid decarboxylase, and the protein tyrosine phosphatase-like molecule IA-2 are found in most patients with type 1 diabetes and are now established markers for the clinical diagnosis and the preclinical phase of this disease. The development of islet autoantibodies and diabetes is influenced by genetic and environmental factors, and the detection and characterization of islet autoantibodies in euglycemic members of affected families identifies some individuals who have a markedly elevated risk for type 1 diabetes. This ability to accurately predict diabetes risk in non-diabetic subjects will prove very useful for targeted recruitment of participants of interventional studies aimed at preventing the progression to type 1 diabetes in subjects at risk. PMID- 15826926 TI - Diabetes-related antibodies in adult diabetic patients. AB - Islet autoimmunity is made evident by the appearance of islet-cell antibodies directed against insulin (IAA), glutamic acid decarboxylase (GADA), protein tyrosine phosphatase IA-2 (IA-2Ab) and other autoantigens. IAA and IA-2Ab are predominantly detected in childhood type 1 diabetes mellitus (T1DM), while frequency of GADA is not affected by age. In adult-onset T1DM patients, GADA is the immune marker of higher diagnostic sensitivity. In adult diabetic patients not requiring insulin treatment for at least 6 months after diagnosis, GADA identifies the so-called latent autoimmune diabetes in adults (LADA). In over 80% of cases, LADA patients develop insulin dependency within a few years after the diagnosis and have an increased risk for the development of other organ-specific autoimmune diseases. High GADA titers identify a subgroup of LADA patients with low body mass index (BMI), low C-peptide levels and increased frequency of T1DM related HLA class II haplotypes. GADA assay should be offered to every diabetic patient, and in cases of positivity screening for other autoimmune diseases should be carried out. PMID- 15826927 TI - Gonadal antibodies interfering with female reproduction. AB - While the involvement of anti-ovarian antibodies (AOAs) is highly likely, yet still controversial, in patients with patent premature ovarian failure (POF), it is even more difficult--for several reasons--to ascertain the clinical significance of these antibodies in patients without obvious ovarian failure. First, AOAs form a heterogeneous group of antibodies recognizing several different antigenic targets such as granulosa and thecal cells, zona pellucida, oocyte cytoplasm, corpus luteum, as well as gonadotrophins and their receptors. Second, the detection of AOAs in various clinical situations does not readily imply a causal relationship between these antibodies and impaired ovarian function. Third, diagnostic tools for detecting AOAs and their molecular targets have to be improved to yield more reliable data and allow a better comprehension of the pathophysiology of AOAs. Preliminary results with immunosuppressive therapy in selected AOA patients have been encouraging, but randomized trials have to be performed. PMID- 15826928 TI - Obstetric complications due to autoantibodies. AB - Autoimmune diseases are most common and most active in young women; it is therefore not uncommon for obstetricians and physicians to encounter pregnant women with these conditions, and knowledge of the potential maternal, foetal and neonatal complications is essential for good clinical management. The most common maternal autoimmune endocrine conditions in pregnancy are insulin-dependent diabetes mellitus and thyroid disease. Other relatively common non-endocrine autoimmune conditions include systemic lupus erythematosus and anti-phospholipid syndrome. Much rarer autoimmune conditions include autoimmune thrombocytopenia, rheumatoid arthritis, myasthenia gravis and Addison's disease. In this chapter, we discuss autoimmune endocrine conditions and briefly mention some non-endocrine conditions of particular importance. PMID- 15826929 TI - Evaluation of risk factors for nasopharyngeal carcinoma in high-risk nasopharyngeal carcinoma families in Taiwan. AB - A study of nasopharyngeal carcinoma (NPC) families with two or more affected members was conducted in Taiwan (265 families with 2,444 individuals, 502 affected and 1,942 unaffected) to determine the association between NPC and potential etiologic factors in NPC high-risk families. Similar to results from a previous case-control study in Taiwan, Guangdong salted fish consumption during childhood, exposure to wood, and betel nut consumption were all associated with elevated NPC risk using conditional logistic regression, although these associations were not as strong as in the case-control study possibly due to shared environment among family members. Risk associated with cumulative wood exposure and salted fish consumption before age 10 was stronger in families with early NPC age-onset [odds ratio (OR(wood)), 5.10; 95% confidence interval (95% CI), 1.50-17.34; OR(fish), 3.94; 95% CI, 1.47-10.55] or three or more affected members (OR(wood), 4.41; 95% CI, 1.58-12.30; OR(fish), 4.27; 95% CI, 1.10-16.47). In contrast, a tendency for elevated risk was noted for betel nut use in late age onset families (OR, 2.44; 95% CI, 1.16-5.13) and the CYP2E1 c2 allele in families with less than three affected members (OR, 2.06; 95% CI, 1.04-3.35). Risk estimates associated with these exposures were similar when the analyses were restricted to EBV-seropositive subjects. To better adjust for degree of relationship among family members and residual genetic correlations, we also calculated ORs using a variance components model. The results from the two methods were similar indicating that the risk estimates from conditional logistic regression were unbiased. PMID- 15826930 TI - Bolus transit patterns in healthy subjects: a study using simultaneous impedance monitoring, videoesophagram, and esophageal manometry. AB - Impedance monitoring (Imp) measures bolus transit. Combining Imp with manometry (EM) allows the effect of contractile patterns on transit to be assessed. The objective of this study is to identify bolus transit patterns in normal subjects, correlate Imp findings with the gold standard barium esophagram (Ba), and compare bolus transit with concomitant EM findings. Simultaneous Ba-Imp-EM was performed for 2 min in 15 normal volunteers (women, 11; age, 43 yr). Combined impedance pressure sites were 5, 10, 15, 20 cm above the lower esophageal sphincter (LES). Boluses (10 ml) of 45% barium mixed with 0.9% NaCl were swallowed at > or = 20-s intervals (5-6 swallows/subject). Imp and Ba showed three bolus transit patterns, and the two methods were in agreement on the pattern type in 97% (83/86) of swallows. Normal bolus transit was found in 73% (61/83), and each had normal peristalsis and contraction amplitude. Stasis in the proximal esophagus occurred in 7 of 83 swallows despite normal manometric parameters in 4 of 7 swallows. Retrograde escape of a residue of incompletely cleared bolus from just above the LES to the site 5 cm above occurred in 14 of 83 swallows. Retrograde escape was triggered by the next swallow, occurred despite normal manometric parameters, and did not occur if the swallow interval was >30 s. In 55% (47/86) of swallows, air accumulated in the distal esophagus and persisted there for a mean of 3.6 s until cleared into the stomach. We conclude that impedance monitoring is a valid transit test and describe bolus transit patterns in normal subjects for comparison with patients with esophageal motility disorders. PMID- 15826931 TI - Toll-like receptor-4 is required for intestinal response to epithelial injury and limiting bacterial translocation in a murine model of acute colitis. AB - Inflammatory bowel disease (IBD) arises from a dysregulated mucosal immune response to luminal bacteria. Toll-like receptor (TLR)4 recognizes LPS and transduces a proinflammatory signal through the adapter molecule myeloid differentiation marker 88 (MyD88). We hypothesized that TLR4 participates in the innate immune response to luminal bacteria and the development of colitis. TLR4-/ and MyD88-/- mice and littermate controls were given 2.5% dextran sodium sulfate (DSS) for 5 or 7 days followed by a 7-day recovery. Colitis was assessed by weight loss, rectal bleeding, and histopathology. Immunostaining was performed for macrophage markers, chemokine expression, and cell proliferation markers. DSS treatment of TLR4-/- mice was associated with striking reduction in acute inflammatory cells compared with wild-type mice despite similar degrees of epithelial injury. TLR4-/- mice experienced earlier and more severe bleeding than control mice. Similar results were seen with MyD88-/- mice, suggesting that this is the dominant downstream pathway. Mesenteric lymph nodes from TLR4-/- and MyD88 /- mice more frequently grew gram-negative bacteria. Altered neutrophil recruitment was due to diminished macrophage inflammatory protein-2 expression by lamina propria macrophages in TLR4-/- and MyD88-/- mice. The similarity in crypt epithelial damage between TLR4-/- or MyD88-/- and wild-type mice was seen despite decreased epithelial proliferation in knockout mice. TLR4 through the adapter molecule MyD88 is important in intestinal response to injury and in limiting bacterial translocation. Despite the diversity of luminal bacteria, other TLRs do not substitute for the role of TLR4 in this acute colitis model. A defective innate immune response may result in diminished bacterial clearance and ultimately dysregulated response to normal flora. PMID- 15826932 TI - Signal-transduction pathways that regulate visceral smooth muscle function. III. Coupling of muscarinic receptors to signaling kinases and effector proteins in gastrointestinal smooth muscles. AB - Stimulation of muscarinic M3 and M2 receptors on gastrointestinal smooth muscle elicits contraction via activation of G proteins that are coupled to a diverse set of downstream signaling pathways and effector proteins. Many studies suggest a canonical excitation-contraction coupling pathway that includes activation of phospholipases, production of inositol 1,4,5-trisphosphate and diacylglycerol, release of calcium from the sarcoplasmic reticulum, activation of L-type calcium channels, and activation of nonselective cation channels. These events lead to elevated intracellular calcium concentration, which activates myosin light chain kinase to phosphorylate and activate myosin II thus causing contraction. In addition, muscarinic receptors are coupled to signaling pathways that modulate the effect of activator calcium. The Rho/Rho kinase pathway inhibits myosin light chain phosphatase, one of the key steps in sensitization of the contractile proteins to calcium. Phosphatidylinositol 3-kinases and Src family tyrosine kinases are also activated by muscarinic agonists. Src family tyrosine kinases regulate L-type calcium and nonselective cation channels. Src activation also leads to activation of ERK and p38 MAPKs. ERK MAPKs phosphorylate caldesmon, an actin filament binding protein. P38 MAPKs activate phospholipases and MAPKAP kinase 2/3, which phosphorylate HSP27. HSP27 may regulate cross-bridge function, actin filament formation, and actin filament attachment to the cell membrane. In addition to the well-known role of M3 muscarinic receptors to regulate myoplasmic calcium levels, the integrated effect of muscarinic activation probably also includes signaling pathways that modulate phospholipases, cyclic nucleotides, contractile protein function, and cytoskeletal protein function. PMID- 15826933 TI - Events at the host-microbial interface of the gastrointestinal tract. II. Role of the intestinal epithelium in pathogen-induced inflammation. AB - An immense number of bacteria reside within the intestinal lumen. The task of appropriately identifying and responding to microbial threats lies primarily with the single layer of cells that line the intestinal tract. Intestinal epithelial cells have developed a number of strategies aimed at identifying microorganisms and eliciting the appropriate inflammatory response. The pathogen recognition mechanisms and the signaling and inflammatory events that ensue within the intestine are the focus of this review. PMID- 15826934 TI - Immediate early genes of glucocorticoid action on the developing intestine. AB - Prior studies have demonstrated that glucocorticoid hormones elicit functional maturation of the small intestine as evidenced by their ability to induce increases in the expression of various digestive hydrolases, such as sucrase isomaltase and trehalase. However, these increases have a lag time of approximately 24 h, suggesting that they are secondary effects of hormone action. To identify candidate primary response genes, we performed microarray analysis on pooled RNA from jejunums of untreated postnatal day 8 mouse pups and from littermates who earlier received dexamethasone 2 h. Fluorescent dye-labeled samples were hybridized in quadruplicate to glass-spotted cDNA microarrays containing 15,000 cDNA clones from the National Institute of Aging cDNA clone set. Analysis of the resulting signals using relatively stringent criteria identified 66 transcripts upregulated and 36 downregulated by 2 h of glucocorticoid treatment. Among the upregulated transcripts, the magnitude of the increase detected by microarray ranged from 1.4- to 16-fold. Selected mRNAs from throughout the range were subsequently analyzed by Northern blot analysis. Of 11 mRNAs chosen all were confirmed, and there was a strong correlation between the magnitude of the increase observed from the microarray analysis and from Northern blot analysis. Additional time points showed that these transcripts peaked between 2 and 6 h and had returned to baseline by 24 h. Gene ontology analysis showed pleiotropic effects of dexamethasone on the developing intestine and pointed to genes in the development category as being likely candidates for mediation of glucocorticoid-induced maturation of intestinal function. PMID- 15826935 TI - Identification of spectrin-like repeats required for high affinity utrophin-actin interaction. AB - Most studies aimed at characterizing the utrophinactin interaction have focused on the amino-terminal tandem calponin homology domain. However, we recently reported evidence suggesting that spectrin-like repeats of utrophin also participate in binding to actin. Here we expressed several recombinant fragments encoding the utrophin amino-terminal domain alone or in combination with various numbers of spectrin-like repeats. We further quantitatively characterized the actin binding properties of each recombinant utrophin fragment using a high-speed sedimentation assay. To evaluate the capacity of each protein to stabilize actin filaments, we compared the effect of utrophin recombinant fragments and full length utrophin on 6-propionyl-2-(N,N-dimethylamino)naphthalene actin depolymerization. Our results suggest that, whereas the amino-terminal domain is essential for primary interaction between utrophin and actin, spectrin-like repeats have additive effects on the affinity and stoichiometry of binding. Our data indicate that the amino-terminal domain and first 10 consecutive spectrin like repeats recapitulate the actin binding activity of full-length utrophin more faithfully than the amino-terminal domain alone. These findings support the model for lateral association of utrophin along the actin filament and provide the molecular basis for designing the most effective utrophin "mini-genes" for treatment of dystrophinopathies. PMID- 15826936 TI - Free cholesterol-loaded macrophages are an abundant source of tumor necrosis factor-alpha and interleukin-6: model of NF-kappaB- and map kinase-dependent inflammation in advanced atherosclerosis. AB - Two key features of atherosclerotic plaques that precipitate acute atherothrombotic vascular occlusion ("vulnerable plaques") are abundant inflammatory mediators and macrophages with excess unesterified, or "free," cholesterol (FC). Herein we show that FC accumulation in macrophages leads to the induction and secretion of two inflammatory cytokines, tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6). The increases in TNF-alpha and IL-6 mRNA and protein were mediated by FC-induced activation of the IkappaB kinase/NF kappaB pathway as well as activation of MKK3/p38, Erk1/2, and JNK1/2 mitogen activated protein kinases (MAPK). Activation of IkappaB kinase and JNK1/2 was needed for the induction of both cytokines. However, MKK3/p38 signaling was specifically involved in TNF-alpha induction, and Erk1/2 signaling was required for IL-6. Most interestingly, activation of all of the signaling pathways and induction of both cytokines required cholesterol trafficking to the endoplasmic reticulum (ER). The CHOP branch of the unfolded protein response, an ER stress pathway, was required for Erk1/2 activation and IL-6 induction. In contrast, one or more other ER-related pathways were responsible for activation of p38, JNK1/2, and IkappaB kinase/NF-kappaB and for the induction of TNF-alpha. These data suggest a novel scenario in which cytokines are induced in macrophages by endogenous cellular events triggered by excess ER cholesterol rather than by exogenous immune cell mediators. Moreover, this model may help explain the relationship between FC accumulation and inflammation in vulnerable plaques. PMID- 15826937 TI - The affinity of the dynein microtubule-binding domain is modulated by the conformation of its coiled-coil stalk. AB - The microtubule-binding domain (MTBD) of dynein is separated from the AAA (ATPase with any other activity) core of the motor by an approximately 15-nm stalk that is predicted to consist of an antiparallel coiled coil. However, the structure of this coiled coil and the mechanism it uses to mediate communication between the MTBD and ATP-binding core are unknown. Here, we sought to identify the optimal alignment between the hydrophobic heptad repeats in the two strands of the stalk coiled coil. To do this, we fused the MTBD of mouse cytoplasmic dynein, together with 12-36 residues of its stalk, onto a stable coiled-coil base provided by Thermus thermophilus seryl-tRNA synthetase and tested these chimeric constructs for microtubule binding in vitro. The results identified one alignment that yielded a protein displaying high affinity for microtubules (2.2 microM). The effects of mutations applied to the MTBD of this construct paralleled those previously reported (Koonce, M. P., and Tikhonenko, I. (2000) Mol. Biol. Cell 11, 523-529) for an intact dynein motor unit in the absence of ATP, suggesting that it resembles the tight binding state of native intact dynein. All other alignments showed at least 10-fold lower affinity for microtubules with the exception of one, which had an intermediate affinity. Based on these results and on amino acid sequence analysis, we hypothesize that dynein utilizes small amounts of sliding displacement between the two strands of its coiled-coil stalk as a means of communication between the AAA core of the motor and the MTBD during the mechanochemical cycle. PMID- 15826938 TI - Rapid in vitro assembly dynamics and subunit turnover of FtsZ demonstrated by fluorescence resonance energy transfer. AB - We have developed an assay for the assembly of FtsZ based on fluorescence resonance energy transfer (FRET). We mutated an innocuous surface residue to cysteine and labeled separate pools with fluorescein (donor) and tetramethylrhodamine (acceptor). When the pools were mixed and GTP was added, assembly produced a FRET signal that was linearly proportional to FtsZ concentration from 0.7 microm (the critical concentration (C(c))) to 3 microm. At concentrations greater than 3 microm, an enhanced FRET signal was observed with both GTP and GDP, indicating additional assembly above this second C(c). This second C(c) varied with Mg(2+) concentration, whereas the 0.7 microm C(c) did not. We used the FRET assay to measure the kinetics of initial assembly by stopped flow. The data were fit by the simple kinetic model used previously: monomer activation, a weak dimer nucleus, and elongation, although with some differences in kinetic parameters from the L68W mutant. We then studied the rate of turnover at steady state by pre-assembling separate pools of donor and acceptor protofilaments. When the pools were mixed, a FRET signal developed with a half-time of 7 s, demonstrating a rapid and continuous disassembly and reassembly of protofilaments at steady state. This is comparable with the 9-s half-time for FtsZ turnover in vivo and the 8-s turnover time of GTP hydrolysis in vitro. Finally, we found that an excess of GDP caused disassembly of protofilaments with a half-time of 5 s. Our new data suggest that GDP does not exchange into intact protofilaments. Rather, our interpretation is that subunits are released following GTP hydrolysis, and then they exchange GDP for GTP and reassemble into new protofilaments, all on a time scale of 7 s. The mechanism may be related to the dynamic instability of microtubules. PMID- 15826939 TI - The Pro33 isoform of integrin beta3 enhances outside-in signaling in human platelets by regulating the activation of serine/threonine phosphatases. AB - Integrin beta(3) is polymorphic at residue 33 (Leu(33) or Pro(33)), and the Pro(33)-positive platelets display enhanced aggregation, P-selectin secretion, and shorter bleeding times. Because outside-in signaling is critical for platelet function, we hypothesized that the Pro(33) variant provides a more efficient signaling than the Leu(33) isoform. When compared with Pro(33)-negative platelets, Pro(33)-positive platelets demonstrated significantly greater serine/threonine phosphorylation of extracellular signal-regulated kinase (ERK2) and myosin light chain (MLC) but not cytoplasmic phospholipase A2 upon thrombin induced aggregation. Tyrosine phosphorylation of integrin beta(3) and the adaptor protein Shc was no different in the fibrinogen-engaged platelets from both genotypes. The addition of Integrilin (alpha(IIb)beta(3)-fibrinogen blocker) or okadaic acid (serine/threonine phosphatase inhibitor) dramatically enhanced ERK2 and MLC phosphorylation in the Pro(33)-negative platelets when compared with Pro(33)-positive platelets, suggesting that integrin engagement during platelet aggregation activates serine/threonine phosphatases. The phosphatase activity of myosin phosphatase (MP) that dephosphorylates MLC is inactivated by phosphorylation of the myosin binding subunit of MP at Thr(696), and aggregating Pro(33)-positive platelets exhibited an increased Thr(696) phosphorylation of MP. These studies highlight a role for the dephosphorylation events via the serine/threonine phosphatases during the integrin outside-in signaling mechanism, and the Leu(33) --> Pro polymorphism regulates this process. Furthermore, these findings support a mechanism whereby the reported enhanced alpha granule secretion in the Pro(33)-positive platelets could be mediated by an increased phosphorylation of MLC, which in turn is caused by an increased phosphorylation and subsequent inactivation of myosin phosphatase. PMID- 15826940 TI - Backbone-methylated analogues of the principle receptor binding region of human parathyroid hormone. Evidence for binding to both the N-terminal extracellular domain and extracellular loop region. AB - We have used backbone N-methylations of parathyroid hormone (PTH) to study the role of these NH groups in the C-terminal amphiphilic alpha-helix of PTH (1-31) in binding to and activating the PTH receptor (P1R). The circular dichroism (CD) spectra indicated the structure of the C-terminal alpha-helix was locally disrupted around the methylation site. The CD spectra differences were explained by assuming a helix disruption for four residues on each side of the site of methylation and taking into account the known dependence of CD on the length of an alpha-helix. Binding and adenylyl cyclase-stimulating data showed that outside of the alpha-helix, methylation of residues Asp30 and Val31 had little effect on structure or activities. Within the alpha-helix, disruption of the structure was associated with increased loss of activity, but for specific residues Val21, Leu24, Arg25, and Leu28 there was a dramatic loss of activities, thus suggesting a more direct role of these NH groups in correct P1R binding and activation. Activity analyses with P1R-delNT, a mutant with its long N-terminal region deleted, gave a different pattern of effects and implicated Ser17, Trp23, and Lys26 as important for its PTH activation. These two groups of residues are located on opposite sides of the helix. These results are compatible with the C terminal helix binding to both the N-terminal segment and also to the looped-out extracellular region. These data thus provide direct evidence for important roles of the C-terminal domain of PTH in determining high affinity binding and activation of the P1R receptor. PMID- 15826941 TI - WAVE3-mediated cell migration and lamellipodia formation are regulated downstream of phosphatidylinositol 3-kinase. AB - WAVE3 is a member of the WASP/WAVE family of protein effectors of actin reorganization and cell movement. The precise role of WAVE3 in cell migration and its regulation, however, have not been elucidated. Here we show that endogenous WAVE3 was found to be concentrated in the lamellipodia at the leading edge of migrating MDA-MB-231 cells. Platelet-derived growth factor (PDGF) treatment induced lamellipodia formation as well as two-dimensional migration of cells in the wound-closure assay and chemotactic migration toward PDGF in three dimensional migration chambers. Knockdown of WAVE3 expression by RNA interference prevented the PDGF-induced lamellipodia formation and cell migration. Treatment of cells with LY294002, an inhibitor of phosphatidylinositol 3-kinase (PI3K), also abrogated the PDGF-induced lamellipodia formation and cell migration, suggesting that PI3K may be required for WAVE3 activity. WAVE3 and the PI3K regulatory subunit, p85, were found to interact in a yeast two-hybrid screen, which was confirmed through co-immunoprecipitation. The WAVE3-p85 interaction was mediated by the N-terminal region of WAVE3 and the C-terminal SH2 domain of p85. These results imply that the WAVE3-mediated migration in MDA-MB-231 cells via lamellipodia formation is activated downstream of PI3K and induced by PDGF. The findings of the WAVE3-p85 partnership also suggest a potential regulatory role for p85 in WAVE3-dependent actin-cytoskeleton reorganization and cell migration. PMID- 15826942 TI - Dual effect of tamoxifen on arterial KCa channels does not depend on the presence of the beta1 subunit. AB - Tamoxifen has been reported to directly activate large conductance calcium activated potassium (KCa) channels through the KCa beta1 subunit, suggesting a cardio-protective role of this compound. The present study using knock-out (KO) mice for the KCa channel beta1 subunit was aimed at understanding the molecular mechanisms of the effects of tamoxifen on arterial smooth muscle KCa channels. Single channel studies were conducted in excised patches from cerebral artery myocytes from both wild-type and KO animals. The present data demonstrated that tamoxifen can inhibit arterial KCa channels due to a major decrease in channel open probability (P(o)), a mechanism different from the reduction in single channel amplitude reported previously and also observed in the present work. A tamoxifen-induced decrease in P(o) was present in arterial KCa channels from both wild-type and beta1 KO animals. This inhibition was concentration-dependent and partially reversible with a half-maximal concentration constant IC(50) of 2.6 microm. The effect of tamoxifen was actually dual Single channel kinetic analysis showed that tamoxifen shortens both mean closed time and mean open time; the latter is probably due to an intermediate duration voltage-independent blocking mechanism. Thus, tamoxifen block would predominate when KCa channel P(o) is >0.1 0.2, limiting the maximum P(o), whereas a leftward shift in voltage or Ca(2+) activation curves can be observed for P(o) values lower than those values. This dual effect of tamoxifen appears to be independent of the beta1 subunit. The molecular specificity of tamoxifen, or eventually other xenoestrogen derivatives, for the KCa channel beta1 subunit is uncertain. PMID- 15826943 TI - Receptor epitope usage by an interleukin-5 mimetic peptide. AB - The cyclic peptide AF17121 is a library-derived antagonist for human interleukin 5 (IL5) receptor alpha (IL5Ralpha) and inhibits IL5 activity. Our previous results have demonstrated that the sixth arginine residue of the peptide is crucial for the inhibitory effect and that several acidic residues in the N- and C-terminal regions also make a contribution, although to a lesser extent (Ruchala, P., Varadi, G., Ishino, T., Scibek, J., Bhattacharya, M., Urbina, C., Van Ryk, D., Uings, I., and Chaiken, I. (2004) Biopolymers 73, 556-568). However, the recognition mechanism of the receptor has remained unresolved. In this study, AF17121 was fused to thioredoxin by recombinant DNA techniques and examined for IL5Ralpha interaction using a surface plasmon resonance biosensor method. Kinetic analysis revealed that the dissociation rate of the peptide.receptor complex is comparable with that of the cytokine.receptor complex. The fusion peptide competed with IL5 for both biological function and interaction with IL5Ralpha, indicating that the binding sites on the receptor are shared by AF17121 and IL5. To define the epitope residues for AF17121, we defined its binding footprint on IL5Ralpha by alanine substitution of Asp(55), Asp(56), Glu(58), Lys(186), Arg(188), and Arg(297) of the receptor. Marked effects on the interaction were observed in all three fibronectin type III domains of IL5Ralpha, in particular Asp(55), Arg(188), and Arg(297) in the D1, D2, and D3 domains, respectively. This footprint represents a significant subset of that for IL5 binding. The fact that AF17121 mimics the receptor binding capability of IL5 but antagonizes biological function evokes several models for how IL5 induces activation of the multisubunit receptor system. PMID- 15826944 TI - Exosome-dependent trafficking of HSP70: a novel secretory pathway for cellular stress proteins. AB - The heat shock proteins (HSPs) are a family of intracellular proteins found in all eukaryotes and prokaryotes. Their functions are well characterized and are central to maintaining cellular homeostasis and in promoting cell survival in response to stressful cellular conditions. However, several studies provide evidence that specific members of the HSP family might be secreted via an unidentified exocytotic pathway. Here we show that exosomes, small membrane vesicles that are secreted by numerous cell types, contribute to the release of HSP70 from human peripheral blood mononuclear cells (PBMCs) in both basal and stress-induced (heat shock at 40 or 43 degrees C for 1 h) states. HSP70 release from PBMCs is independent of the common secretory pathway because Brefeldin A, an inhibitor of the classical protein transport pathway, did not block HSP70 release. Furthermore, we show that HSP70 release from PBMCs does not occur via a lipid raft-dependent pathway, because treatment with methyl-beta-cyclodextrin, a raft-disrupting drug, had no affect on HSP70 release. To examine whether exosomes contributed to HSP70 release from PBMCs, exosomes were purified from PBMC cultures, and exosomal number and HSP70 content were determined. We demonstrate that although heat shock does not influence the exosomal secretory rate, the HSP70 content of exosomes isolated from heat shocked PBMCs is significantly higher than control. These data identify a novel secretory pathway by which HSP70 can be actively released from cells in both the basal and stress-induced state. PMID- 15826945 TI - Involvement of GADD153 and cardiac ankyrin repeat protein in hypoxia-induced apoptosis of H9c2 cells. AB - Oxidative stress is the main cause of cardiac injury during ischemia/reperfusion but the molecular mechanism for this process is unclear. In this study, it was found that hypoxia induces apoptosis in rat embryonic heart-derived H9c2 cells leading to the induction of GADD153, which is an apoptosis-related gene. Therefore, this study addressed the molecular role of GADD153 in hypoxia-induced apoptosis. The stable or inducible overexpression of GADD153 sensitized the H9c2 cells to apoptotic cell death. The results suggest that the transactivation domain of the GADD153 might be responsible for this cell execution and play a role in the nucleoplasmic localization of GADD153. The cells transiently transfected with the antisense GADD153 were more resistant to hypoxia-induced apoptosis than the vector control cells. Furthermore, GADD153 transcriptionally down-regulated the expression of the cardiac ankyrin repeat protein gene (CARP), which is a nuclear transcriptional co-factor that negatively regulates the expression of the cardiac gene. The ectopic expression of CARP in H9c2 cells increased the resistance to hypoxia-induced apoptosis. These results suggest that GADD153 overexpression and the concomitant down-regulation of CARP might have a causative role in the apoptotic cell injury of hypoxic H9c2 cells. PMID- 15826946 TI - Calcium-dependent changes in the flexibility of the regulatory domain of troponin C in the troponin complex. AB - With the recent advances in structure determination of the troponin complex, it becomes even more important to understand the dynamics of its components and how they are affected by the presence or absence of Ca(2+). We used NMR techniques to study the backbone dynamics of skeletal troponin C (TnC) in the complex. Transverse relaxation-optimized spectroscopy pulse sequences and deuteration of TnC were essential to assign most of the TnC residues in the complex. Backbone amide (15)N relaxation times were measured in the presence of Ca(2+) or EGTA/Mg(2+). T(1) relaxation times could not be interpreted precisely, because for a molecule of this size, the longitudinal backbone amide (15)N relaxation rate due to chemical shift anisotropy and dipole-dipole interactions becomes too small, and other relaxation mechanisms become relevant. T(2) relaxation times were of the expected magnitude for a complex of this size, and most of the variation of T(2) times in the presence of Ca(2+) could be explained by the anisotropy of the complex, suggesting a relatively rigid molecule. The only exception was EF-hand site III and helix F immediately after, which are more flexible than the rest of the molecule. In the presence of EGTA/Mg(2+), relaxation times for residues in the C-domain of TnC are very similar to values in the presence of Ca(2+), whereas the N-domain becomes more flexible. Taken together with the high flexibility of the linker between the two domains, we concluded that in the absence of Ca(2+), the N-domain of TnC moves independently from the rest of the complex. PMID- 15826947 TI - Galpha12/13-mediated production of reactive oxygen species is critical for angiotensin receptor-induced NFAT activation in cardiac fibroblasts. AB - Angiotensin II (Ang II) activates multiple signaling pathways leading to hyperplasia of cardiac fibroblasts. Reactive oxygen species (ROS) produced by Ang II stimulation are assumed to play pivotal roles in this process. Here, we show that ROS mediate Ang II-induced activation of nuclear factor of activated T cells (NFAT) in rat cardiac fibroblasts. Ang II-induced NFAT activation was suppressed by diphenyleneiodonium (an NADPH oxidase inhibitor), dominant negative (DN)-Rac, DN-p47(phox), and an inhibitor of Galpha(12/13) (Galpha(12/13)-specific regulator of G protein signaling domain of p115RhoGEF, p115-regulator of G protein signaling (RGS)). Stimulation of Ang II receptor increased the intracellular ROS level in a Rac- and p47(phox)-dependent manner. Because p115-RGS suppressed Ang II-induced Rac activation, Ang II receptor-coupled Galpha(12/13) mediated NFAT activation through ROS production by Rac activation. Ang II-induced nuclear translocation of the green fluorescent protein (GFP)-tagged amino-terminal region of NFAT4 (GFP-NFAT4) was suppressed by p115-RGS or BAPTA but not by diphenyleneiodonium. The expression of constitutively active (CA)-Galpha(12/13), CA-G translocation alpha(13), or CA-Rac increased the nuclear of GFP-NFAT4. These results suggest that NFAT activity is regulated by both Ca(2+)-dependent and ROS dependent pathways. Furthermore, activation of c-Jun NH(2)-terminal kinase (JNK) induced by Ang II stimulation is required for NFAT activation because Ang II induced NFAT activation was inhibited by SP600125, a selective JNK inhibitor. These results indicate that Ang II stimulates the nuclear translocation and activation of NFAT by integrated pathways including the activation of Galpha(12/13), Rac, NADPH oxidase, and JNK and that Galpha(12/13)-mediated ROS production is essential for NFAT transcriptional activation. PMID- 15826948 TI - Are STATS arginine-methylated? AB - Transcription factors of the STAT (signal transducer and activator of transcription) family are important in signal transduction of cytokines. They are subject to post-translational modification by phosphorylation on tyrosine and serine residues. Recent evidence suggested that STATs are methylated on a conserved arginine residue within the N-terminal region. STAT arginine methylation has been described to be important for STAT function and loss of arginine methylation was discussed to be involved in interferon resistance of cancer cells. Here we provide several independent lines of evidence indicating that the issue of arginine methylation of STATs has to be reassessed. First, we show that treatment of melanoma and fibrosarcoma cells with inhibitors used to suppress methylation (N-methyl-2-deoxyadenosine, adenosine, dl-homocysteine) had profound and rapid effects on phosphorylation of STAT1 and STAT3 but also on p38 and Erk signaling cascades which are known to cross-talk with the Jak/STAT pathway. Second, we show that anti-methylarginine antibodies did not precipitate specifically STAT1 or STAT3. Third, we show that mutation of Arg(31) to Lys led to destabilization of STAT1 and STAT3, implicating an important structural role of Arg(31). Finally, purified catalytically active protein arginine methyltransferases (PRMT1, -2, -3, -4, and -6) did not methylate STAT proteins, and cotransfection with PRMT1 did not affect STAT1-controlled reporter gene activity. Taken together, our data suggest the absence of arginine methylation of STAT1 and STAT3. PMID- 15826949 TI - Identification of a myosin VII-talin complex. AB - Myosin VII (M7) plays a role in adhesion in both Dictyostelium and mammalian cells where it is a component of a complex of proteins that serve to link membrane receptors to the underlying actin cytoskeleton. The nature of this complex is not fully known, prompting a search for M7-binding proteins. Co immunoprecipitation experiments reveal that Dictyostelium M7 (DdM7) interacts with talinA, an actin-binding protein with a known role in cell-substrate adhesion. No additional proteins are observed in the immunoprecipitate, indicating that the interaction is direct. The N-terminal region of the DdM7 tail that lies between the region of predicted coil and the first MyTH4 domain is found to harbor the talinA binding site. Localization experiments reveal that talinA does not serve as a membrane receptor for DdM7 and vice versa. These findings reveal that talinA is a major DdM7 binding partner and suggest that their interaction induces a conformational change in each that, in combination with membrane receptor binding, promotes the assembly of a high avidity receptor complex essential for adhesion of the cell to substrata. PMID- 15826950 TI - Repression of interleukin-5 transcription by the glucocorticoid receptor targets GATA3 signaling and involves histone deacetylase recruitment. AB - Glucocorticoids are the mainstay of asthma therapy and mediate the repression of a number of cytokine genes, such as Interleukin (IL)-4, -5, -13, and granulocyte macrophage colony-stimulating factor (GM-CSF), which are central to the pathogenesis of asthmatic airway inflammation. The glucocorticoid receptor (GR) mediates repression by a number of diverse mechanisms. We have previously suggested that one such repressive activity is by direct binding of GR to elements within the GM-CSF enhancer that are recognized by the nuclear factor of activated T cells.activator protein 1 (NF-AT.AP-1) complex. We reasoned that, because many cytokine genes activated in asthma are transcriptionally regulated by the recruitment of this complex to DNA, their binding sites might provide a target for GR to mediate its repressive effects. Here, we show that transcriptional repression of the Interleukin-5 gene involves recruitment of GR to a DNA region located within the IL-5 proximal promoter, which is bound by NF AT and AP-1 proteins. GR recruitment had a profound effect upon the activation capacity of GATA3, which has a binding site close to the NF-AT.AP-1 domain in both IL-5 and IL-13 promoters. Repression by GR involves co-repressor recruitment, because treatment of transfected cells with the deacetylase inhibitor trichostatin A caused a partial relief of repression. Additionally, repression could be augmented by co-transfection of cells with a histone deacetylase (HDAC1). These data suggest that the local recruitment of GR causes repression by inhibiting transcriptional activation by GATA3, a key tissue specific determinant of expression of Th2 cytokines. PMID- 15826951 TI - Myosin II regulatory light chain is required for trafficking of bile salt export protein to the apical membrane in Madin-Darby canine kidney cells. AB - BSEP, MDR1, and MDR2 ATP binding cassette transporters are targeted to the apical (canalicular) membrane of hepatocytes, where they mediate ATP-dependent secretion of bile acids, drugs, and phospholipids, respectively. Sorting to the apical membrane is essential for transporter function; however, little is known regarding cellular proteins that bind ATP binding cassette proteins and regulate their trafficking. A yeast two-hybrid screen of a rat liver cDNA library identified the myosin II regulatory light chain, MLC2, as a binding partner for BSEP, MDR1, and MDR2. The interactions were confirmed by glutathione S transferase pulldown and co-immunoprecipitation assays. BSEP and MLC2 were overrepresented in a rat liver subcellular fraction enriched in canalicular membrane vesicles, and MLC2 colocalized with BSEP in the apical domain of hepatocytes and polarized WifB, HepG2, and Madin-Darby canine kidney cells. Expression of a dominant negative, non-phosphorylatable MLC2 mutant reduced steady state BSEP levels in the apical domain of polarized Madin-Darby canine kidney cells. Pulse-chase studies revealed that Blebbistatin, a specific myosin II inhibitor, severely impaired delivery of newly synthesized BSEP to the apical surface. These findings indicate that myosin II is required for BSEP trafficking to the apical membrane. PMID- 15826952 TI - Mechanism of transcription factor recruitment by acidic activators. AB - Many transcriptional activators are intrinsically unstructured yet display unique, defined conformations when bound to target proteins. Target-induced folding provides a mechanism by which activators could form specific interactions with an array of structurally unrelated target proteins. Evidence for such a binding mechanism has been reported previously in the context of the interaction between the cancer-related c-Myc protein and the TATA-binding protein, which can be modeled as a two-step process in which a rapidly forming, low affinity complex slowly converts to a more stable form, consistent with a coupled binding and folding reaction. To test the generality of the target-induced folding model, we investigated the binding of two widely studied acidic activators, Gal4 and VP16, to a set of target proteins, including TATA-binding protein and the Swi1 and Snf5 subunits of the Swi/Snf chromatin remodeling complex. Using surface plasmon resonance, we show that these activator-target combinations also display bi phasic kinetics suggesting two distinct steps. A fast initial binding phase that is inhibited by high ionic strength is followed by a slow phase that is favored by increased temperature. In all cases, overall affinity increases with temperature and, in most cases, with increased ionic strength. These results are consistent with a general mechanism for recruitment of transcriptional components to promoters by naturally occurring acidic activators, by which the initial contact is mediated predominantly through electrostatic interactions, whereas subsequent target-induced folding of the activator results in a stable complex. PMID- 15826953 TI - Main chain hydrogen bond interactions in the binding of proline-rich gluten peptides to the celiac disease-associated HLA-DQ2 molecule. AB - Binding of peptide epitopes to major histocompatibility complex proteins involves multiple hydrogen bond interactions between the peptide main chain and major histocompatibility complex residues. The crystal structure of HLA-DQ2 complexed with the alphaI-gliadin epitope (LQPFPQPELPY) revealed four hydrogen bonds between DQ2 and peptide main chain amides. This is remarkable, given that four of the nine core residues in this peptide are proline residues that cannot engage in amide hydrogen bonding. Preserving main chain hydrogen bond interactions despite the presence of multiple proline residues in gluten peptides is a key element for the HLA-DQ2 association of celiac disease. We have investigated the relative contribution of each main chain hydrogen bond interaction by preparing a series of N-methylated alphaI epitope analogues and measuring their binding affinity and off-rate constants to DQ2. Additionally, we measured the binding of alphaI gliadin peptide analogues in which norvaline, which contains a backbone amide hydrogen bond donor, was substituted for each proline. Our results demonstrate that hydrogen bonds at P4 and P2 positions are most important for binding, whereas the hydrogen bonds at P9 and P6 make smaller contributions to the overall binding affinity. There is no evidence for a hydrogen bond between DQ2 and the P1 amide nitrogen in peptides without proline at this position. This is a unique feature of DQ2 and is likely a key parameter for preferential binding of proline rich gluten peptides and development of celiac disease. PMID- 15826954 TI - Distinct amino acid residues may be involved in coactivator and ligand interactions in hepatocyte nuclear factor-4alpha. AB - Hepatocyte nuclear factor-4 (HNF-4) is a transcription factor of the nuclear hormone receptor superfamily that is constitutively active without the addition of exogenous ligand. Crystallographic analysis of the HNF-4alpha and HNF-4gamma ligand binding domains (LBDs) demonstrated the presence of endogenous ligands that may act as structural cofactors for HNF-4. It was also proposed by crystallographic studies that a combination of ligand and coactivator might be required to lock the receptor in its active state. We previously showed that mutations in amino acid residues Ser-181 and Met-182 in H3, Leu-219 and Leu-220 and Arg-226 in H5, Ileu-338 in H10, and Ileu-346 in H11, which line the LBD pocket in HNF-4alpha and come in contact with the ligand, impair its transactivation potential. In the present study, physical and functional interaction assays were utilized with two different coactivators, PGC-1 and SRC 3, to address the role of coactivators in HNF-4 function. We show that the integrity of the hinge (D) domain of HNF-4alpha and the activation function (AF) 2 activation domain region are critical for coactivation. Surprisingly, a different mode of coactivation is observed among the LBD point mutants that lack transcriptional activity. In particular, coactivation is maintained in mutants Ser-181, Arg-226, and Ile-346 but is abolished in mutants Met-182, Leu-219, and Ile-338. Physical interactions confirm this pattern of activation, implying that distinct amino acid residues may be involved in coactivator and ligand interactions, although some residues may be critical for both functions. Our results provide evidence and expand predictions based on the crystallographic data as to the role of coactivators in HNF-4alpha constitutive transcriptional activity. PMID- 15826955 TI - The canonical intrinsic mitochondrial death pathway has a non-apoptotic role in signaling lens cell differentiation. AB - The mitochondrial cell death pathway is known for its role in signaling apoptosis. Here, we describe a novel function for the mitochondrial cell death pathway in signaling initiation of differentiation in the developing lens. Most remarkably, we induced lens cell differentiation by short-term exposure of lens epithelial cells to the apoptogen staurosporine. Activation of apoptosis-related pathways induced lens epithelial cells to express differentiation-specific markers and to undergo morphogenetic changes that led to formation of the lens like structures known as lentoids. The fact that multiple stages of differentiation are expressed at a single stage of development in the embryonic lens made it possible to precisely determine the timing of expression of proteins associated with the apoptotic pathway. We discovered that there was high expression in the lens equatorial epithelium (the region of the lens in which differentiation is initiated) of pro-apoptotic molecules such as Bax and Bcl-x(S) and release of cytochrome c from mitochondria. Furthermore, we found significant caspase-3-like activity in the equatorial epithelium, yet this activity was far lower than that associated with lens cell apoptosis. These apoptotic pathways are likely regulated by the concurrent expression of prosurvival molecules, including Bcl-2 and Bcl-x(L); phosphorylation of Bad; and high expression of inhibitor of apoptosis proteins chicken IAP1, IAP3, and survivin. This finding suggests that prosurvival pathways allow pro-apoptotic molecules to function as molecular switches in the differentiation process without tipping the balance toward apoptosis. We call this process apoptosis-related Bcl-2- and caspase-dependent (ABC) differentiation. PMID- 15826956 TI - Proprioception and palisade endings in extraocular eye muscles. AB - Palisade endings occur only in extraocular muscles, and their function is unknown. They form a cuff of nerve terminals around the tips of muscle fibers. We describe here the advantages of using antibodies to a synaptosomal-associated protein (SNAP-25) to study properties of palisade endings in man, monkey, and rat. The stain can be combined readily with other immunofluorescence procedures, and results suggest that the synapses of palisade endings do not bind alpha bungarotoxin (i.e., are not motor), nor do they contain substance P. These double labeling data support the hypothesis that palisade endings are non-nociceptive sensory receptors, and could serve a proprioceptive function. With SNAP-25 immunolabeling, palisade endings were identified in the rat for the first time. Thus, palisade endings appear to be present in all vertebrate extraocular muscles studied to date. Their apparent universality, which contrasts with the more variable manifestation of extraocular muscle spindles and Golgi tendon organs, would be expected if proprioceptive feedback is necessary to the function of the ocular motor system, and if palisade endings are the critical proprioceptive structure. PMID- 15826957 TI - SEE++: a biomechanical model of the oculomotor plant. AB - The consequences of changes in the oculomotor system on the three-dimensional eye movements are difficult to grasp. Although changes to the rectus muscles can still be approximately understood with simplified geometric models, this approach no longer works with the oblique muscles. It is shown how SEE++, a biomechanical model of the oculomotor plant that was built on the ideas of Miller and Robinson (1984) can improve the understanding of the effects of changes to the oblique eye muscles. By displaying only selected muscles, and by illustrating the relative contribution of these muscles through color-coding the bulb surface, the functional properties of the oblique muscles can be presented in a much clearer way. Investigating the effects of a hyperactive inferior oblique muscle shows that this type of model can help to clarify the functional cause of a pathology, which can otherwise be unclear, even for common pathologies. PMID- 15826958 TI - Human angular vestibulo-ocular reflex axis disconjugacy: relationship to magnetic resonance imaging evidence of globe translation. AB - Magnetic resonance imaging (MRI) demonstrates that the lateral rectus pulley shifts 0.5 mm inferiorly relative to the medial rectus in 20 degrees upgaze, but 0.5 mm superiorly in 20 degrees downgaze, whereas the globe translates 0.7 mm nasally in adduction and 0.2 mm nasally in abduction. If pulleys influence ocular kinematics, these effects would predict disconjugate alterations of the yaw vestibulo-ocular reflex (VOR) rotational axes. Binocular eye and head movements were recorded using three-dimensional search coils in 8 humans (age 24 +/- 2 years, mean +/- SE) undergoing directionally randomized, transient, whole-body yaw (2800 degrees /s2 peak) in darkness while fixating straight ahead, as well as +/- 18 degrees vertically. Eye and head rotational velocity axes were expressed as quaternions in Listing coordinates. In the initial 70 ms, the ocular axis varied with vertical gaze by one-quarter the angle of target elevation, but this effect summed significantly with a disconjugate effect of horizontal duction. In central gaze, the mean adducting eye (AD) rotational axis tilted 3.4 +/- 0.8 degrees forward relative to the head axis, while that of the abducting eye (AB) tilted 0.6 +/- 0.8 degrees backward. In downgaze, the AD rotational axis tilted 8.6 +/- 1.0 degrees forward, and AB 5.7 +/- 1.2 degrees forward. In upgaze, the AD rotational axis tilted backward by 0.1 +/- 0.7 degrees, and AB backward 3.4 +/ 0.9 degrees. We suggest that nasal globe translation relative to the fixed trochlea produces binocular extorsion accounting for yaw VOR axis disconjugacy, and thus a horizontal duction dependence in VOR rotational axis summating with classic dependence of VOR axis on vertical gaze. Confirmation of predicted duction-dependent VOR disconjugacy supports the idea that rectus pulleys influence kinematics for all eye movements. PMID- 15826959 TI - Human angular vestibulo-ocular reflex initiation: relationship to Listing's law. AB - An ideal vestibulo-ocular reflex (VOR) generates ocular rotations compensatory for head motion. During visually guided movements, Listing's law (LL) constrains eye rotation to axes in Listing's plane (LP). Recently, it has been reported that the VOR axis is not collinear with the rotation axis of the head, but is influenced by eye position in the orbit. Elaborate models have been proposed suggesting dynamic neural control of the VOR axis. By examining the variability and time course of changes in VOR axis orientation, we sought to test plausibility of these models. Binocular LPs were defined in eight humans. The VOR was evoked by a highly repeatable, transient, whole-body yaw rotation in darkness at peak acceleration 2800 deg/s2. Immediately prior to rotation, subjects regarded targets at eye level, 20 degrees up, or 20 degrees down. Eye and head positions were expressed in LP coordinates for comparison with LL. Eye position generally followed head position and departed LP when the head axis tilted out of LP. In the velocity domain the VOR axis tilted 28 +/- 9% of the change in vertical eye position, but there was significant intrasubject variation (14% to 41%). This roughly "quarter-angle" behavior began with the earliest detectable VOR. Given the brief latency and marked interindividual variability of the eye position dependence of the VOR rotational axis, and the small deviation of the VOR from LL in the position domain, it is speculated that this behavior is largely due to orbital mechanics interacting with the basic neural commands that initiate the VOR. PMID- 15826960 TI - Binocular coordination in fore/aft motion. AB - Stabilization of images on the fovea during either fore/aft translation of a subject or fore/aft movement of a visual target in front of a stationary observer imposes complex geometrical requirements that depend upon the eccentricity of the object of interest with respect to the eyes. Each eye needs to be rotated independently with varying proportions of conjugate (version) and disconjugate (vergence) eye movements to maintain fixation of the target. Here, we describe binocular coordination in the early response to translational movements of normal subjects along their naso-occipital axis. We recorded the responses evoked by small (about 4 cm), abrupt (about 0.7 g), fore/aft translations in four normal subjects while they viewed a near target. In the forward and backward starting positions the target was 15 or 10.5 cm away, respectively. Each subject was tested with the target centered between the eyes, aligned on the right eye, and placed to the right of the right eye by approximately 3 cm. The three conditions differed only in the lateral eccentricity of the target, yet the geometrical requirements for image stabilization are very different: pure vergence, one eye still, or mostly version. We found that the eye-movement responses closely matched what was needed for visual stabilization of the target, though responses to stimuli calling for divergence were less accurate than those for convergence. The latency of these responses ranged from 40 to 65 ms and achieved about 80% of the ideal response by 90 to 100 ms after the onset of the stimulus. Next, we asked whether these eye movements were generated by the vestibular system or by high-level strategies for image stabilization, such as pursuit. Thus, in a second set of experiments we used the mean profile of fore?aft body motion computed for each subject to drive a small visual target across the same distances and in the same eccentricities used during body translations. We found that visually driven responses had longer latencies (by at least 80 ms, ranging from 144 to 155 ms) and slower dynamics (with significantly lower peak eye velocities), highlighting the different subsystems producing the two types of responses. Saccades were also an important component of the response to both visual and vestibular stimuli, less frequent during the centered-target configuration and more frequent during viewing of eccentric targets. Visual stimuli evoked saccadic corrections more often and at shorter latencies than did vestibular stimuli. Both smooth and saccadic eye movements were appropriately disconjugate and their pattern depended on whether the eyes were converging or diverging. PMID- 15826961 TI - Vestibular responses to sound. AB - Research into vestibular responses to sound has evolved in four stages. The first, largely the work of Tullio in the 1920s, involved inspection of the eye, head, and postural responses to sound of alert animals with surgical fenestrae into various parts of the bony labyrinth. The second, begun in 1964 by Bickford and his group and continued by our group and then by others in the last 10 years, involves the measurement of evoked myogenic potentials to air-conducted and bone conducted clicks and tones in normal humans. The third, begun by Mikaelian at about the same time as Bickford and continued by McCue, our group, and others, involves electrophysiological recordings of primary vestibular afferent neuron responses to sound in anesthetized animals. The fourth involves measurements of vestibulo-ocular responses to sound in humans with the Tullio phenomenon. It was begun by Minor and his group in 1998 with the observation that sound-induced nystagmus in humans, the Tullio phenomenon, aligned with the rotation axis of the superior semicircular canal. They then showed a defect in the temporal bone between the apex of the superior semicircular canal and the middle cranial fossa, which was the cause of most, if not all, cases of sound-induced nystagmus. Here some of the key observations made in each of these four stages are reviewed. PMID- 15826962 TI - Angular and linear vestibulo-ocular responses in humans. AB - A new technique is introduced to measure linear and angular vestibulo-ocular responses in three dimensions. Using a three-dimensional motion platform, human subjects underwent whole-body rotations and translations. Eye movements were measured with an infrared video recording device and/or with scleral search coils. Subjects were tested with sinusoidal stimulation and impulses under light and dark conditions. The results show that for sinusoidal stimulation, torsion compensatory eye movements (roll stimulation) have a low gain compared to horizontal (yaw) and vertical (pitch) compensatory eye movements. With impulses, we reliably assessed gain and delay for rotations in yaw, pitch, and roll. Under this stimulus condition the gain for roll (torsion eye movements) was also low compared to yaw and pitch (horizontal and vertical eye movements). For translations, the gain of eye-movement responses varied between 0.7 and 1 in the light. In the dark, responses were lower and more variable. PMID- 15826963 TI - Residual torsion following ocular counterroll. AB - A recent study on static ocular counterroll suggested the existence of residual torsion (RT): when healthy subjects repositioned their head to the upright position after sustained static tilt, eye position differed from the original ocular torsion measured prior to the static head tilt. Our experiments aimed at further characterizing this phenomenon. Using a three-dimensional motorized turntable, healthy human subjects (n = 8) were rotated quasi-statically (0.05 deg/s2, 2 deg/s velocity plateau reached after 40 s) from the upright position about the naso-occipital axis. Three full whole-body rotations were completed while subjects fixed upon a blinking laser dot straight ahead in otherwise complete darkness. Three-dimensional eye movements were recorded with modified dual search coils (wires exiting inferiorly). Torsional position of the right eye at consecutive upright body positions was analyzed. The torsional eye position before the beginning of the chair rotation was defined as zero torsion. On average, the right eye was intorted by 1.3 degrees or extorted by 2.0 degrees after the first full chair rotation in the clockwise or counterclockwise direction, respectively. These torsional offset values of the right eye did not significantly change after the two subsequent full chair rotations. We conclude that RT observed after static ocular counterroll is the result of static hysteresis, that is, a position lag of the eye, which depends on the direction of head roll. The fact that residual torsion did not further increase after the first rotation cycle emphasizes that RT is a static rather than a dynamic phenomenon. PMID- 15826964 TI - Adaptation of VOR to Coriolis stimulation. AB - The vestibulo-ocular reflex (VOR) is normally characterized by the gain and phase of slow-phase velocity (SPV) relative to the stimulus velocity. Although this is perfectly satisfactory for steady-state sinusoidal oscillations about a single axis, it is less useful when applied to transient responses. The well-known decay of nystagmus following a step change of head velocity approximately follows a double exponential, with an initial amplitude (A), a long time constant (tau), and an adaptation time constant (tau(a)). We have developed a means of representing the transient response for a complex head velocity stimulus as experienced during high-speed artificial gravity (AG) experiments. When a subject, lying supine on a rotating horizontal platform, makes a yaw head movement of amplitude theta, the vertical semicircular canals experience a step in angular velocity. The pitch stimulus is equal to the change in the component of the centrifuge angular velocity (omega(c)) aligned with the interaural axis, and gives rise to a vertical VOR. The magnitude of the step change is omega(c) sin theta. The SPV is approximated by an exponential decay of amplitude A and single time constant tau, and then normalized relative to this stimulus step. MATLAB scripts filter the raw eye position data to remove noise, blinks, and saccades, differentiate the signal, and remove fast phases to obtain SPV. The amplitude of the fitted SPV exponential is divided by omega(c) sin theta to obtain the normalized SPV. A and tau are shown to behave differently as subjects adapt to repeated head movements of different amplitudes. PMID- 15826965 TI - The role of gravity in adaptation of the vertical angular vestibulo-ocular reflex. AB - The gain of the vertical angular vestibulo-ocular reflex (aVOR) was adapted in side-down and prone positions in two monkeys and tested in four planes: left /right-side down; forward/backward; and two intermediate planes that lie approximately in the planes of the vertical semicircular canal pairs, left anterior/right posterior (LA/RP) and right anterior/left posterior (RA/LP). Gain changes, expressed as a percent of preadapted values, were plotted as a function of head orientation in the planes of tilt, and fitted with sinusoids to obtain the gravity-dependent (amplitude) and gravity-independent (bias) components of adaptation. Gravity-dependent gain changes were always maximal when tested in a plane that included the head orientation in which the aVOR gain had been adapted. Changes were minimal when the head was tilted in a plane orthogonal to the plane of adaptation, and were smaller but still significant when tested in the two intermediate planes. Gravity-independent VOR gain changes were uniform over all planes of head tilt. Thus, the gravity-dependent and gravity-independent components could be separated experimentally. The aVOR gain changes from the head tilts in different directions were utilized to reconstruct the gain changes in three dimensions. They formed a continuous surface, which peaked in and around the position of adaptation. These studies support the postulate that gain adaptation has both gravity-independent and gravity-dependent components, and further show that these gain changes have a three-dimensional structure. These results are similar to those in humans, indicating that the gravity-dependent adaptation of the aVOR is likely to be a common phenomenon across species. PMID- 15826966 TI - Vestibular cortical area in the periarcuate cortex: its afferent and efferent projections. AB - Vestibular input to the periarcuate cortex in the Japanese monkey was examined by analyzing laminar field potentials evoked by electrical stimulation of the vestibular nerve. Vestibular-evoked potentials consisted of early-positive and late-negative potentials and early-negative and late-positive potentials in the superficial and deep layers of the cortex, respectively. They were distributed bilaterally in the periarcuate cortex around the junction of the spur and the arcuate sulcus. This vestibular-projecting area corresponded to the periarcuate area where retrogradely-labeled corticovestibular neurons were distributed after the injection of a tracer into the vestibular nuclei. Comparison of the vestibular-projection area with the distribution of smooth pursuit-related neurons in the same monkey revealed that such neurons existed in the vestibular projecting area of the periarcuate cortex. PMID- 15826967 TI - Saccular projections in the human cerebral cortex. AB - The cerebral cortical areas processing saccular information were investigated in human subjects using the fMRI method and loud clicks, which selectively activate the saccule. The results were compared with previous vestibular evoked potential (VEP) studies in anesthetized patients following vestibular nerve stimulation. Nine normal subjects participated in fMRI studies. By comparing the cortical areas activated by a click at 85 dB (auditory activation) with those activated by 102 dB (auditory plus saccular activation), the following cortical areas were selectively activated by saccular stimulation: intraparietal sulcus, frontal eye fields, prefrontal cortex, and postcentral gyrus, in addition to insula, supplementary motor area, and anterior and posterior cingulate cortex. Previous VEP studies also revealed similar activation areas by vestibular nerve stimulation with latencies at 6 ms, suggesting that the shortest pathways for activation of cerebral cortical neurons from the labyrinth are trisynaptic, with a relay in the thalamus. The activated areas are also consistent with results in previous studies using caloric stimulation, which primarily activates horizontal semicircular canals. These results suggest that canal and otolith information is processed largely by similar cortical areas in humans. Multiple cortical areas activated by these studies suggest that these areas are involved in different aspects of processing vestibular information. The saccular projections to the prefrontal and frontal cortex suggest that these areas are involved in planning motor synergies to counteract loss of equilibrium. PMID- 15826968 TI - Sensorimotor transformation for visually guided saccades. AB - Visually guided movements require the brain to perform a sensorimotor transformation. The key to understanding this transformation is to understand the different roles of the superior colliculus (SC) and cerebellum (CB). The SC has a three-layered structure. Cells in the top layer have visual, but not motor, responses. However, cells in the deeper layers have both visual and motor responses. Thus, for a long time it was thought that the SC encoded both the retinal location of a sensory stimulus and the desired change in eye movement needed to acquire it. However, copious evidence has accumulated that shows that the SC encodes only the retinal location of a visual target, and not the movement needed to foveate it. Thus, the information needed to make accurate movements must come from another part of the brain, which is proposed to be the cerebellum. Here it is shown how the cerebellum could perform the sensorimotor transformation. PMID- 15826969 TI - Saccades to moving targets. AB - The metrics and dynamics of saccades to stationary and moving targets were observed in monkeys (Macaca mulatta). To isolate the effects of target speed on the saccade from contributions of smooth pursuit, saccade velocity was corrected for intrasaccadic pursuit velocity on a trial-by-trial basis prior to analysis. The effects of presaccadic retinal error and target speed on the saccadic velocity profile were determined by analyzing the partial correlations computed as a function of time after saccade onset. The main results are: (1) Saccade amplitude is determined not only by the retinal error sampled before the saccade, but also by the speed of the target during the latency period. (2) The dynamics of saccades, even if compensated for smooth-pursuit components, differ between forward- and backward-moving targets. (3) Whereas the presaccadic retinal error affects the eye velocity throughout the saccade, target speed has no effect before peak velocity. These results are discussed in the context of current models of saccade generation and their physiological substrates, in particular the role of the cerebellum in the local feedback loop. PMID- 15826970 TI - Visual motion perception at the time of saccadic eye movements and its relation to spatial mislocalization. AB - The same retinal image motion can be produced by a variety of combinations of eye and target motion. In natural conditions, extraretinal information disambiguates retinal information for motion perception. By controlling the timing of visual motion with respect to saccades, it was possible to appraise the roles for motion perception of the signal related to saccades occurring in the vicinity of visual motion. When a visual motion was seen before or after a saccade, its perceived direction was biased in the direction opposite to the saccade. The magnitude of the bias depended on the timing of the visual motion with respect to the saccade and the meridian of the visual motion. The bias appears to be independent of the deformation of visual space reported to occur before and after saccades. PMID- 15826971 TI - Context-specific adaptation of saccade gain is enhanced with rest intervals between changes in context state. AB - Dual-state adaptation of motor responses has been known for some time. A more recent development is a form of dual-state adaptation known as "context-specific adaptation," which was explored through the use of saccade gain adaptation. In this model, two different adapted saccade gains are associated with two different states of a context cue, and the gain switches between the two adapted states when the context cue changes state. Such adaptation is imposed by alternating context/adaptation states over the course of an adaptation session. Here, vertical eye position as a context cue for adaptation of horizontal saccade gain is used: gain increase is induced with the eyes up 10 degrees, and gain decrease with the eyes down 10 degrees. This context cue is not very effective: there is interference between context/adaptation conditions such that gain-decrease adaptation with eyes down transfers to the eyes-up (gain-increase) context. It was hypothesized that the juxtaposition in time of the alternating adaptation states exacerbated this interference. In order to test this, one-minute rest breaks were inserted between each change in context/adaptation state. The resulting context-specific adaptation improved dramatically: gain-increase and gain-decrease adaptations were more rapid and more complete. This resembles consolidation of motor learning, which, however, occurs over much longer time spans (hours rather than minutes). Thus, the results may reflect the operation of a novel "short-term" motor consolidation process. PMID- 15826972 TI - Cognitive processes in saccade generation. AB - The analysis of saccades offers an opportunity to study a number of different cognitive processes, such as visuospatial attention, working memory, and volitional conflict. A study of saccades in patients with visuospatial hemineglect, who performed a visual search task, showed repeated fixations on targets previously discovered, yet they often failed to retain the information that a particular target had previously been discovered. High-resolution structural brain scanning showed that this abnormality was due either to a lesion in the right intraparietal sulcus or the right inferior frontal lobe. Detailed analysis of the scanpaths suggested that the former location was associated with an accumulating impairment in remapping target locations across saccades or impaired memory of previously inspected target locations, whereas the latter location was more consistent with a failure to inhibit responses to rightward locations. When combined with a spatial bias to the right, such deficits might explain why many neglect patients often reexamine rightward targets, at the expense of items to their left. The functions of the supplementary eye field (SEF), in the medial frontal lobe, in relation to saccade generation are controversial. A series of studies in a patient with a focal lesion of the right SEF has indicated an important role for the SEF in the rapid self-control of saccadic eye movements and in set-switching (i.e., implementing control in situations of response conflict when ongoing saccadic plans have to be changed rapidly), rather than monitoring errors. In a recent fMRI study of normal subjects, it was shown that the SEF is involved in implementing the resolution of any volitional conflict, whereas other presupplementary motor areas are involved in the generation of volitional plans and processing volitional conflict. PMID- 15826973 TI - Role of the posterior parietal cortex in the initiation of saccades and vergence: right/left functional asymmetry. AB - This study explored in humans the role of the posterior parietal cortex (PPC) in saccades, vergence, and combined saccade-vergence movements by means of transcranial magnetic stimulation (TMS). TMS was applied to the right PPC at 80 ms, 90 ms, or 100 ms after target onset in experiment 1, and to the left PPC in experiment 2. Control experiments were also run in which TMS was applied over the primary motor cortex at 90 ms after target onset. Relative to no-TMS trials, TMS over the right PPC prolonged significantly the latency of almost all eye movements (saccades in either direction, convergence, divergence, and components of combined eye movements). Such latency increase was significant mostly when TMS was delivered 90 ms after target onset. In contrast, TMS of the left PPC increased the latency only for saccades to right, convergence, and convergence combined with rightward saccades; latency increase occurred for all time windows of TMS deliver (80, 90, or 100 ms after target onset). TMS over the vertex had no effect on the latency for any type of eye movement. TMS of either the left or the right PPC or of the motor cortex did not alter the accuracy of any type of eye movement. Thus, the effects of TMS on latency are time-, area-, and eye-movement specific. We suggest that the right PPC is involved primarily in the processing of fixation disengagement, whereas the left PPC participates in the initiation of eye movements via different spatial selective mechanisms that concern exclusively targets to the right and/or to near. PMID- 15826974 TI - Readout of higher-level processing in the discharge of superior colliculus neurons. AB - The discharge of neurons in the deeper layers of the superior colliculus (SC) was studied while monkeys performed two visual discrimination tasks that required different amounts of cognitive processing. In a search paradigm the animal's task was to saccade to the location of an odd-colored stimulus located in an array of distractors of uniform color (pop-out visual search). The visual stimuli remained on the screen as the discrimination process distinguished target from distractors. In a choice response task the color of a central cue signaled which stimulus from a previously presented array of colored stimuli was to be the target of a saccade. The stimulus array was turned off well before the central cue was presented. Most neurons showed activity aligned on both the visual input and the motor response in single-target tasks. Many of these same neurons showed additional discharge that was correlated with the required higher-level decision processes in both of these more natural visual tasks. In the case of pop-out search the SC has been shown to be functionally involved in the decision processes. The cue-aligned activity in SC in the choice response task is surprising because no transient visual stimulus appeared in the response field of the neuron. PMID- 15826975 TI - Synaptic inputs and their pathways from fixation and saccade zones of the superior colliculus to inhibitory burst neurons and pause neurons. AB - The caudal part of the superior colliculus (SC) plays an important role in the generation of saccades, whereas the rostral part of the SC is considered to be involved in visual fixation. The present study was performed to determine neural connections from the rostral and caudal parts of the SC to inhibitory burst neurons (IBNs) and pause neurons (PNs) in the nucleus raphe interpositus in the anesthetized cat, and to reveal the functional role of the rostral SC on eye movements. The intracellular potentials from IBNs and PNs were recorded, and the effects of stimulation of the SC on these neurons were analyzed. The results show that IBNs receive monosynaptic excitation from the contralateral caudal SC, and disynaptic inhibition from the ipsilateral caudal SC via contralateral IBNs. In addition, IBNs receive disynaptic inhibition from the rostral part of the SC on either side via inhibitory interneurons other than IBNs. Intracellular recording from PNs revealed that they receive convergent excitation from the rostral parts of the bilateral superior colliculi and that the rostral SC inhibits IBNs on both sides via PNs. The neural connections determined in this study support the functional independence of the rostral SC and are consistent with the notion that the "fixation zone" is localized in the rostral SC. These results show that the fixation zone in the rostral SC may suppress the initiation of bilateral saccades via pause neurons. PMID- 15826976 TI - Initiation and suppression of saccades by the frontal eye field in the monkey. AB - After a saccadic eye movement occurs to an interesting object appearing in the visual field, visual fixation holds its image on the fovea and suppresses saccades to other objects appearing in the visual field. To understand the neural mechanism of visual fixation, the effects of electrical stimulation of the frontal eye field (FEF) on the generation of electrically evoked saccades (Esacs) and the suppression of saccades in trained monkeys were investigated. When the properties of the electrically evoked suppression of visually guided (Vsacs) and memory-guided saccades (Msacs) were examined, two types of suppression were found. Stimulation of a wide area of the FEF suppressed only ipsiversive Vsacs and Msacs at stimulus intensities lower than those for eliciting Esacs, whereas stimulation of a localized area of the FEF suppressed the initiation of both Vsacs and Msacs in any direction during and approximately 50 ms after stimulation. However, neither stimulation affected the vector of these saccades. The thresholds for suppression were usually less than 50 microA. The most effective stimulation timing for the suppression of ipsiversive and contraversive Vsacs was approximately 40 to 50 ms before saccade onset. Therefore, suppression occurred in the efferent pathway for Vsacs at the premotor rather than the motoneuronal level, most likely in the superior colliculus and/or the paramedian pontine reticular formation. The results suggest that the suppression in the FEF may play a role in maintaining visual fixation by suppressing the generation of saccades. PMID- 15826977 TI - Cortical mechanisms of saccade generation from execution to decision. AB - Several cortical areas are involved in the control of ocular motor behavior, but little is known about the cognitive processes occurring prior to saccade triggering. The use of functional brain imaging allows a better understanding of the networks involved specifically in distinct processes of saccade generation. The use of sophisticated visual stimulation paradigms in functional imaging studies suggests that the decision process of where to look when faced with visual alternatives is subserved by a prefrontoparietal network, including the dorsolateral prefrontal cortex. PMID- 15826978 TI - The role of the human dorsolateral prefrontal cortex in ocular motor behavior. AB - The dorsolateral prefrontal cortex (DLPFC) is involved in the preparation of saccadic eye movements. Lesion studies and functional magnetic resonance imaging (fMRI) studies suggest that the human DLPFC is located in area 46 of Brodmann. The DLPFC has direct connections with the main cortical ocular motor areas, that is with the frontal eye field (FEF) and the supplementary eye field (SEF) in the frontal lobe; with several (associative, attentional, and motor) areas in the posterior parietal cortex (PPC), including the parietal eye field (PEF); with the cingulate eye field in the anterior cingulate cortex; and directly downstream with the superior colliculus in the brainstem. Lesion and fMRI studies using the antisaccade paradigm have shown that the DLPFC is involved in the inhibition of unwanted reflexive saccades (triggered toward the target by the PEF), whereas the triggering of correct intentional antisaccades (made in the direction opposite to the target) may depend mainly upon the FEF. The DLPFC also controls short-term spatial working memory involved in memory-guided saccades, as shown by lesion and transcranial magnetic stimulation (TMS) studies. By contrast, medium-term spatial memory (after 25 s) may be controlled by the medial temporal cortex (MTC). Recently, TMS studies have suggested that the transmission of memorized information from the integrative parietal areas (PPC) to the MTC is performed via both an indirect pathway comprising the DLPFC (i.e., transmission in series) and a direct pathway bypassing the DLPFC (i.e., transmission in parallel). Furthermore, the DLPFC is involved in the preparation of predictive saccades (i.e., saccades made before the appearance of an expected target) and saccade sequences, and, therefore, also controls some aspects of temporal working memory. Lastly, the involvement of the DLPFC has recently been reported in tasks comprising a target selection or a directional decision to make for the forthcoming saccade. These different functions suggest that the DLPFC plays a major role in the decisional processes governing ocular motor behavior. PMID- 15826979 TI - Short-latency disparity vergence in humans: evidence for early spatial filtering. AB - Our study was concerned with the disparity detectors underlying the initial disparity vergence responses (DVRs) that are elicited at ultrashort latencies by binocular disparities applied to large images. DVRs were elicited in humans by applying horizontal disparity to vertical square-wave gratings lacking the fundamental (termed here, the "missing fundamental"). In the frequency domain, a pure square wave is composed of odd harmonics--first, third, fifth, seventh, etc. -such that the third, fifth, seventh, etc., have amplitudes that are one-third, one-fifth, one-seventh, etc., that of the first, and the missing fundamental lacks the first harmonic. The patterns seen by the two eyes have a phase difference of one-quarter wavelength, so the disparity of the features and 4n + 1 harmonics (where n = integer) has one sign (crossed or uncrossed), whereas the 4n - 1 harmonics--including the strongest Fourier component (the third harmonic)- has the opposite sign (uncrossed or crossed): spatial aliasing. The earliest DVRs, recorded with the search-coil technique, had minimum latencies of 70 to 80 ms and were generally in the direction of the third harmonic, that is, uncrossed disparities resulted in convergent eye movements. In other experiments on the DVRs, one eye saw a missing fundamental and the other saw a pure sine wave with the contrast and wavelength of the third harmonic but differing in phase by one quarter wavelength. This resulted in short-latency vergence in accordance with matching of the third harmonic. These data all indicate the importance of the Fourier components, consistent with early spatial filtering prior to binocular matching. PMID- 15826980 TI - Initial ocular following in humans depends critically on the fourier components of the motion stimulus. AB - Visual motion is sensed by low-level (energy-based) and high-level (feature based) mechanisms. Our interest is in the motion detectors underlying the initial ocular following responses (OFR) that are elicited at ultrashort latencies by sudden motions of large images. OFR were elicited in humans by applying horizontal motion to vertical square-wave gratings lacking the fundamental. In the frequency domain, a pure square wave is composed of the odd harmonics--first, third, fifth, seventh, etc.--such that the third, fifth, seventh, etc., have amplitudes that are one-third, one-fifth, one-seventh, etc., that of the first, and the missing fundamental stimulus lacks the first harmonic. Motion consisted of successive quarter-wavelength steps, so the features and 4n+1 harmonics (where n = integer) shifted forward, whereas the 4n-1 harmonics--including the strongest Fourier component (the third harmonic)--shifted backward (spatial aliasing). Thus, the net Fourier energy and the non-Fourier features moved in opposite directions. Initial OFR, recorded with the search coil technique, had minimum latencies of 60 to 70 ms and were always in the direction of the third harmonic, for example, leftward steps resulted in rightward OFR. Thus, the earliest OFR were strongly dependent on the motion of the major Fourier component, consistent with mediation by oriented spatiotemporal visual filters as in the well-known energy model of motion detection. Introducing interstimulus intervals of 10 to 100 ms (during which the screen was uniform gray) reversed the initial direction of tracking, consistent with extensive neurophysiological and psychophysical data suggesting that the visual input to the motion detectors has a biphasic temporal impulse response. PMID- 15826981 TI - Role of vestibular signals in the caudal part of the frontal eye fields in pursuit eye movements in three-dimensional space. AB - For accurate visual information about objects of interest moving slowly in three dimensional (3D) space, primates with binocular fields use both frontal smooth pursuit (frontal-pursuit) and vergence eye movements (i.e., depth pursuit) to maintain the images of the objects precisely on the foveae of left and right eyes. Moreover, during head or whole-body movement, both frontal- and depth pursuit systems must interact with the vestibular system to minimize slip of the retinal images that degrades image quality considerably. The caudal part of the frontal eye fields (FEF) contains many frontal-pursuit neurons. Previous studies have shown that a majority of pursuit neurons there discharge for both frontal pursuit and vergence and carry pursuit-in-3D signals. To understand how vestibular inputs interact with pursuit-in-3D signals, three different experiments that examined the nature of vestibular signals in the caudal FEF are described in this review. A majority of caudal FEF pursuit neurons responded to whole-body rotation with preferred directions similar to frontal-pursuit directions and carried frontal gaze (eye-in-space) velocity signals. They were activated in association with adaptive pursuit eye movements induced by cross axis pursuit-vestibular interactions. During fore/aft and right/left translation in complete darkness, they were also modulated with preferred directions of many neurons similar to pursuit-preferred directions. Previous studies showed that caudal FEF pursuit neurons also receive visual signals about target motion. Taken together, these results suggest that the caudal FEF coordinates its various inputs to provide signals for accurate eye-movement-in-space commands. PMID- 15826982 TI - Yellow filters can improve magnocellular function: motion sensitivity, convergence, accommodation, and reading. AB - The magnocellular system plays an important role in visual motion processing, controlling vergence eye movements, and in reading. Yellow filters may boost magnocellular activity by eliminating inhibitory blue input to this pathway. It was found that wearing yellow filters increased motion sensitivity, convergence, and accommodation in many children with reading difficulties, both immediately and after three months using the filters. Motion sensitivity was not increased using control neutral density filters. Moreover, reading-impaired children showed significant gains in reading ability after three months wearing the filters compared with those who had used a placebo. It was concluded that yellow filters can improve magnocellular function permanently. Hence, they should be considered as an alternative to corrective lenses, prisms, or exercises for treating poor convergence and accommodation, and also as an aid for children with reading problems. PMID- 15826983 TI - Tilt psychophysics measured in nonhuman primates. AB - A method to measure tilt psychophysics in nonhuman primates has been developed and tested. Two rhesus monkeys were trained to orient a light bar parallel to the direction of gravity by rotating a small steering wheel attached to the primate chair. After adequate training, they performed this task during motion paradigms that rotated the gravitoinertial force (GIF) in the roll plane. The animals accurately aligned the light bar with the direction of gravity during and following roll tilts. Subtle aspects of perceived roll tilt previously recorded in human subjects were also observed in the monkeys. During centrifugation and linear translation, an interaural inertial force rotates the GIF in the roll plane. Similar to humans, the animals' perceived direction of gravity shifted toward alignment with the GIF during these paradigms, and the dynamics of these shifts demonstrated an interaction between the otolith afferents that transduce the GIF and rotational cues from the semicircular canals. Finally, optokinetic roll stimulation induced an illusion of roll tilt that was dependent on head orientation, similar to results described in human subjects. Taken together, these findings validate the methodology employed to measure tilt psychophysics; the monkeys' responses were qualitatively similar to those described in human subjects and also recapitulated subtle aspects of the perceptual response observed in humans. This new methodology sets the stage for innovative experimental approaches that will help to elucidate the neural mechanisms underlying the perception of spatial orientation. PMID- 15826984 TI - Vestibular memory-contingent whole-body return: brave exocentered dancers. AB - A whole-body, self-driven return from passive rotation (90 degrees to 270 degrees ) test was executed by male and female subjects, and by professional female ballet dancers. To accomplish the return task, subjects were free to use the egocentered reference frame (inversion) or the exocentered one (completion). The results show that with inversion all kinetic parameters were reproduced, whereas the completion performance was highly variable. Although inversion was the default strategy, female subjects used more completions than male subjects, and female dancers still more, although not more accurately. The high variability noted in completion shows a deficiency in integrating vestibular signals for updating the egocentric representation of an external target during passive body rotations, even in dancers. Furthermore, with completions after 180 degrees stimuli, the results suggest that both ego- and exocentered reference frames cannot be used simultaneously. PMID- 15826985 TI - Is an internal model of head orientation necessary for oculomotor control? AB - In order to test whether the control of eye movement in response to head movement requires an internal model of head orientation or instead can rely on directly sensing information about head orientation and movement, perceived gravity was separated from physical gravity to see which dominated the eye-movement response. Internal model theory suggests that the oculomotor response should be driven by perceived, internalized gravity, whereas the direct sensing theory predicts it should always be driven by vestibularly sensed gravity. Subjects lay on an airbed either supine or on their side and were sinusoidally translated along their dorsoventral body axis. The direction of perceived gravity was separated from physical gravity by performing the experiments in a room built on its side with the direction of its "floor" orthogonal to both physical gravity and the subject's translation. The swinging sum of the imposed sinusoidal acceleration with physical gravity was thus in a plane orthogonal to its sum with perceived gravity. Oculomotor responses to these swinging vectors were looked for and responses were found only to the sum of the acceleration with physical gravity, not perceived gravity. It was concluded that an internal model is not used to drive these compensatory eye movements. PMID- 15826986 TI - Expectation of sensory stimulation modulates brain activation during visual motion stimulation. AB - The differential effects of visual hemifield motion stimulation during fixation of a stationary target were compared under two conditions: fixation straight ahead without any further instructions and fixation straight ahead with attention shifted to the "dark hemifield." Data from nine right-handed volunteers revealed that striate and extrastriate right hemispheric visual areas exhibited larger activations during left hemifield motion stimulation when attention was shifted to the right dark hemifield. Montreal Neurological Institute (MNI) coordinates (26, -98, -4) of the additional clusters activated in the latter condition corresponded best to the kinetic occipital region, which is known to process both shape and motion information, and to parts of area V3 posterior to V3A, which has been shown repeatedly to mediate motion perception. A simple computational model of transhemispheric visuovisual interaction is proposed. The basic mechanism of this model is a central predictor formed by a feedback loop that detects a mismatch between input to the two hemispheres. Predicted stimulation is then compared with the actual input. If the sensed motion of a visual hemifield is larger than the predicted net motion in the model, activation of the respective neural population is increased; conversely, a smaller actual motion causes less activation. PMID- 15826987 TI - Gaze position corrective eye movements in normal subjects and in patients with vestibular deficits. AB - Eye movements in response to high-acceleration head rotations (thrusts) in the horizontal plane from patients with unilateral (UVD) or bilateral vestibular loss (BVD) were recorded. The rapid, gaze-position corrections (GPCs) that appeared when vestibulo-ocular reflex (VOR) slow phases were undercompensatory were characterized. For comparison, eye movements from normal subjects who were asked to generate saccades in the direction opposite head rotation (in the same direction as slow phases) were recorded. This normal-subject model produced responses with spatial and temporal characteristics similar to those from GPCs in patients as follows: When head rotations were generated actively, compared with passively, gaze-position errors and corresponding GPCs were smaller and occurred earlier. During passively generated head thrusts, GPCs still occurred when head rotations were made in total darkness, though their accuracy decreased as the requirement for maintaining gaze on a specific location in space was relaxed. Time of onset of GPCs was not rigidly tied to head kinematics (peak velocity or peak acceleration). Speeds of GPCs, however, were lower than speeds of similar sized, head-fixed saccades. Finally, during passive and active head thrusts in patients, sustained, high-frequency (20 to 30 Hz) oscillations that appeared as tiny saccades were occasionally observed, one immediately following the other, resembling a compensatory slow-phase response. Taken together, the results suggest that one strategy for overcoming a VOR deficit is to enlist the saccadic system to produce an oculomotor response that is required to compensate for head rotation. This response may come in the form of high-velocity GPCs or smaller amplitude oscillations. PMID- 15826988 TI - Asymmetry of the pitch vestibulo-ocular reflex in patients with cerebellar disease. AB - Responses to pitch head impulses were measured in 15 patients with cerebellar degeneration and downbeat nystagmus and in 5 control subjects, using three-axis search coils. For each subject, response gains were calculated as (1) the ratio of instantaneous vertical eye velocity to pitch head velocity (at 30 ms and at 70 ms into the response), and (2) the ratio of peak eye velocity to peak head velocity. Gains varied more widely among patients. When calculated at 70 ms and using peak values, patients had higher gains for downward pitch, and normal subjects had symmetric gains. At 30 ms, gains were more symmetric in patients, but at that point control subjects actually had lower gains for downward pitch. Thus, the pitch gain ratio (ratio of downward gain to upward gain) was consistently greater in patients than in normal subjects. Because downward impulses were generally faster, eye velocities during downward and upward pitch for equivalent head speeds were also compared. Again, patients had higher gains for downward pitch. These results are consistent with the authors' hypothesis that cerebellar disease results in a higher sensitivity of anterior than posterior semicircular canal pathways, perhaps through loss of inhibition from the flocculus/paraflocculus complex on anterior canal secondary neurons in the vestibular nuclei. The pitch gain asymmetry was larger than, and did not correlate with, the velocity of spontaneous upward drift. This supports the notion that other mechanisms are likely to contribute to downbeat nystagmus in these patients. PMID- 15826989 TI - Acute vestibular neuritis: prognosis based upon bedside clinical tests (thrusts and heaves). AB - We compared bedside tests of vestibulo-ocular function (head thrust and head heave signs) with caloric testing results in 68 patients with acute vestibular neuritis seen at onset and in follow-up for one year. The head thrust and head heave signs each were strong predictors of a decreased probability of recovery, and if both were present, there was a trend for a slower recovery and a further decrease in the probability of recovery. If the head thrust sign was absent, recovery was assured. Our results suggest that careful bedside testing of semicircular canal (head thrust maneuver) and otolith (head heave maneuver) function provides useful information for predicting prognosis in patients with acute vestibular neuritis. PMID- 15826990 TI - Medial vestibular nucleus lesions in Wallenberg's syndrome cause decreased activity of the contralateral vestibular cortex. AB - Three patients with the clinical diagnosis of Wallenberg's syndrome caused by acute unilateral ischemic infarctions, which included the vestibular nucleus in the medullary brain stem and afferent vestibular pathways, were examined by positron emission tomography (PET) during caloric vestibular stimulation. They all had typical signs of vestibular dysfunction such as transient rotatory vertigo with vomiting at the onset, ipsiversive body and ocular lateropulsion, and a complete ocular tilt reaction with tilts of the subjective visual vertical. Compared with healthy volunteers, who show activation in a network of temporoparietal vestibular areas within both hemispheres, especially in the posterior insula and retroinsular region that contains the human homologue of the parietoinsular vestibular cortex (PIVC) in monkeys, the activation pattern of the patients with Wallenberg's syndrome was typically changed. During caloric irrigation of the ear ipsilateral to the side of the lesion, they showed no or significantly reduced activation in the contralateral hemisphere, whereas the activation pattern in the ipsilateral hemisphere appeared "normal." These results are compatible with bilateral ascending vestibular pathways from the vestibular nuclei to the vestibular cortex. The novel finding in all three patients was that the activation patterns were compatible with the assumption that only the crossing fibers from the medial vestibular subnucleus to the contralateral medial longitudinal fascicle were affected, but the ipsilateral vestibular thalamocortical projections via the superior vestibular subnucleus were spared. Thus, the activation pattern in the PET study may reflect the vestibular tonic imbalance within the vestibular nuclei circuitry at the cortical level. PMID- 15826991 TI - Clinical implications of a mathematical model of benign paroxysmal positional vertigo. AB - Benign paroxysmal positional vertigo (BPPV) is the most commonly diagnosed vertigo syndrome. It is caused by movement of detached otoconia within the inner ear (canalithiasis) or otoconia adherent to the cupula (cupulolithiasis). A mathematical model incorporating fluid dynamics of BPPV, which results in the following insights, has been developed recently: (1) The characteristic latency of BPPV is explained by movement of detached otoconia through the ampulla, as pressure caused by moving otoconia is negligible until otoconia enter the narrow duct of the semicircular canal. Typical otoconia move at a rate of 0.2 mm/s, or about 1% of the circumference of the canal each second. (2) Particle-wall interactions can account for the considerable variability in duration and latency of BPPV. (3) Dispersion of a clump of otoconia creates more rather than less nystagmus. Thus, dispersion is not a viable explanation of fatigability. (4) Cupulolithiasis is predicted to cause a far weaker nystagmus than canalithiasis. (5) Inertial effects of treatment maneuvers cause negligible movement of otoconia. PMID- 15826992 TI - Effect of 3,4-diaminopyridine on the postural control in patients with downbeat nystagmus. AB - Downbeat nystagmus (DBN) is a common, usually persistent ocular motor sign in vestibulocerebellar midline lesions. Postural imbalance in DBN may increase on lateral gaze when downbeat nystagmus increases. 3,4-Diaminopyridine (3,4-DAP) has been shown to suppress the slow-phase velocity component of downbeat nystagmus and its gravity-dependent component with concomitant improvement of oscillopsia. Because the pharmacological effect is thought to be caused by improvement of the vestibulocerebellar Purkinje cell activity, the effect of 3,4-DAP on the postural control of patients with downbeat nystagmus syndrome was examined. Eye movements were recorded with the video-based Eyelink II system. Postural sway and pathway were assessed by posturography in lateral gaze in the light and on eye closure. Two out of four patients showed an improvement of the area of postural sway by 57% of control (baseline) on eye closure. In contrast, downbeat nystagmus in gaze straight ahead and on lateral gaze did not benefit in these two patients, implying a specific influence of 3,4-DAP on the vestibulocerebellar control of posture. It was concluded that 3,4-DAP may particularly influence the postural performance in patients with downbeat nystagmus. PMID- 15826993 TI - Evaluating large saccades in patients with brain-stem or cerebellar disorders. AB - Clinicians conventionally test saccades at the bedside by noting the accuracy, initiation time, and speed of large movements, with the patient's head stationary. Partly for methodological reasons, laboratory analysis of saccades has mainly focused on movements of 20 degrees or less. By measuring the velocity waveform of large saccades, it is possible to examine more closely the way in which brain stem and cerebellum guide the eye to the target. Large saccades made by healthy humans show a positively skewed velocity profile. Slow saccades made by patients with brain-stem disorders show a prolonged plateau of low velocity. Some patients with cerebellar disorders may show increased acceleration and deceleration of saccades. Each of these velocity waveforms can be modeled by changing the parameters that describe medium-lead burst neuron firing. In certain other brain-stem and cerebellar disorders, transient decelerations or premature terminations of saccades occur; such velocity waveforms cannot be modeled solely by changing the parameters that describe burst neuron firing. Instead, it is necessary to postulate dysfunction of the mechanism that normally inhibits pontine omnipause neurons, thereby permitting burst neurons to discharge until the saccade is completed. Analysis of large, abnormal saccades calls for application of novel techniques to identify the beginning and end of the saccadic pulse command. PMID- 15826994 TI - Recovery of peripheral versus central nerves identified by saccadic velocity after abducens neuropathy. AB - The abducens is the motor nerve with the most substantial course, both within and outside the brain and it innervates only one muscle. Sixth nerve palsy affords an opportunity to compare recovery after central versus peripheral nerve damage by assessing the dynamics of abduction. Horizontal saccade peak velocities and durations in 14 patients with unilateral peripheral sixth nerve palsies (5 acute, 9 chronic) are compared with those in 5 patients with central sixth nerve palsies (2 acute, 3 chronic) and with those in 10 normal subjects. Acutely, abducting saccades in the paretic eye were slow in both central and peripheral palsies, as anticipated from weakness of the lateral rectus muscle. In chronic central palsies, abducting saccadic velocities remained reduced, but in chronic peripheral palsies, they increased to normal within the limited range of excursion. The chronically damaged peripheral nerve behaves like a high-pass filter in transmitting phasic velocity commands, whereas tonic position commands remain defective, accounting for limited abduction but normal velocities within the range of duction. In chronic central (fascicular) palsies, saccade velocities remain reduced. Impaired conduction from damage to central myelin or axons is more persistent in central palsies, consistent with limited regeneration within the brain. Recording of saccade velocities may aid the distinction of fascicular from peripheral palsies. Saccade speed is repaired in peripheral palsies, probably by remyelination, and perhaps also by central monocular adaptation of innervation selectively to the paretic eye in order to drive both eyes rapidly and simultaneously to a target in the paretic field of motion. PMID- 15826995 TI - Relative atrophy of the flocculus and ocular motor dysfunction in SCA2 and SCA6. AB - Two hereditary ataxia syndromes show distinct profiles of region-specific atrophy and ocular motor deficits. Selective pontine atrophy is associated with slowed saccades in ataxin-2 mutations, and selective floccular atrophy is associated with impaired pursuit and gaze-holding abnormalities in Ca(V)2.1 mutations. Although the flocculus seems to be spared relative to the pons in ataxin-2 mutations, and pursuit and gaze-holding appear to be relatively normal, these can be difficult to assess at the bedside, as corrective saccades are also slow and hard to discern. Here, we show the presence of significant floccular atrophy compared with controls in both ataxin-2 and Ca(V)2.1 mutations, which raises the possibility that abnormalities of smooth pursuit or gaze-holding are present in both conditions. PMID- 15826996 TI - Niemann-Pick type C disease in two affected sisters: ocular motor recordings and brain-stem neuropathology. AB - Two sisters with Niemann-Pick disease type C were examined: the brain in one sister, who had died, was examined, and eye movements in the other, surviving sister were recorded. Ocular motor recordings showed marked slowing of vertical saccades with relative sparing of horizontal saccades, pursuit, and the vestibulo ocular reflex. Neuropathological findings included glial fibrillary lesions in the area of the posterior commissure and neuronal loss in the rostral interstitial nucleus of the MLF with preservation of the interstitial nucleus of Cajal and ocular motor complex. These neuropathologic findings correlate well with our current understanding of the anatomy and physiology of the supranuclear control of vertical gaze. PMID- 15826997 TI - Potential role of anti-GAD antibodies in abnormal eye movements. AB - Glutamic acid decarboxylase (GAD) catalyzes the conversion of glutamic acid to gamma-aminobutyric acid (GABA). Autoantibodies directed against GAD (antiGAD-Ab) have been described in patients with insulin-dependent diabetes mellitus, stiff man syndrome, and in a few patients with progressive cerebellar ataxia. The presence of these autoantibodies suggests an autoimmune pathophysiological mechanism for the neurological manifestations in these disorders. However, the exact role of antiGAD-Ab and GABAergic neurotransmission in the pathogenesis of the neurological manifestations, particularly in progressive cerebellar ataxia, is not fully understood. The cases of two patients with subacute cerebellar ataxia associated with antiGAD-Ab presenting with abnormal eye movements are reported. One patient presented a periodic alternating nystagmus (PAN), whereas the other presented a downbeat nystagmus (DBN) and slow vertical saccades. The potential role of antiGAD-Ab and the resultant GABAergic neurotransmission deficit in oculomotor manifestations is discussed. PMID- 15826998 TI - Dependence of the torsional vestibulo-ocular reflex on the direction of gravity. AB - The torsional vestibulo-ocular reflex (tVOR), which is mainly driven by input from the semicircular canals, has been shown to be augmented by otolith input: its gain is increased in the upright as compared to the supine head orientation. We tested whether otolith input in right/left ear down and upside-down orientation also contributes to the tVOR. In contrast to current models of canal otolith interaction, which predict equal responses in upright and upside-down orientations, we found that the tVOR in the upside-down position is significantly decreased, possibly due to an additive interaction of canal and otolith signals. PMID- 15826999 TI - About the effects of velocity saturation on smooth pursuit. AB - By means of simple simulations and based on experimental results from the literature, it is argued that correct consideration of the well known velocity saturation of the smooth pursuit eye movement system may suggest new insight into some intriguing aspects of this system's behavior. PMID- 15827000 TI - Effects of retinal image slip on modulation of visual vestibulo-ocular reflex during near viewing. AB - During viewing of a near target, the gain of the visually modulated vestibulo ocular reflex (vVOR) exceeds 1.0. We investigated whether retinal image slip contributes to this gain increase by measuring responses during strobe illumination. Subjects showed greater gain values during normal versus strobe illumination of the near target, even during rotation at frequencies above 1.0 Hz, indicating that retinal image velocity information is used to optimize vVOR. PMID- 15827001 TI - Tests of Hering- and Helmholtz-type models for saccade-vergence interactions by comparing visually guided and memory-guided movements. AB - We compared the dynamic properties of memory-guided and visually-guided saccade vergence movements. For memory-guided responses, convergence components were slowed proportionally more than corresponding saccadic components, compared with visually-guided responses. This result is consistent with independent saccadic and vergence systems, and supports a Hering-type model for saccade-vergence interactions. PMID- 15827002 TI - Automatic detection of camera translation in eye video recordings using multiple methods. AB - A concern with video eye movement tracking is that movement of the camera headset relative to the head creates an artifact of eye movement in pupil-detection software. We describe the development of, and compare the results of, three automatic image processing algorithms to measure camera movement. The best of the algorithms has an average accuracy of 1.3 pixels, equivalent to 0.49 deg with our eye tracking system. PMID- 15827003 TI - Comparison of velocity waveforms of eye and head saccades. AB - We compared the velocity waveforms of head and eye-in-head movements during eye head saccades over a range 5-50 degrees. The velocity waveforms of eye-in-head saccades showed a skew ratio (acceleration period/saccade duration) that varied as a function of saccade size and duration. The velocity waveforms of head saccades showed a more constant skew of approximately 0.5 for the range of movements tested. These results suggest independent system controllers for eye and head components of eye-head saccades. PMID- 15827004 TI - Fixation stability in normal children. AB - Fixation stability was found to increase with increasing age (4-15 years) in normal children, but there was no directional preponderance in fixation location. PMID- 15827005 TI - Phase-synchronization decay of fixational eye movements. AB - In nonstationary noisy systems the traditional cross-correlation method may not appropriately detect all cases of interdependencies between coupled systems. The phase-synchronization method was previously found useful in detecting synchronization in several systems. We here applied the phase-synchronization decay to study the synchronization between six combinations of binocular fixational eye movement components. We found that only two components were synchronized: the right and left horizontal with each other and the right and left vertical. Furthermore, the vertical-vertical components were much more synchronized than the horizontal. PMID- 15827006 TI - Human surge linear vestibulo-ocular reflex during tertiary gaze viewing. AB - The otolith-mediated linear vestibulo-ocular reflex (IVOR) was studied in 9 normal humans undergoing transient whole-body surges at 0.5 g peak acceleration while viewing targets eccentrically placed in tertiary positions that combined horizontal and vertical eccentricities at distance of 15, 25, or 50 cm both in darkness and light. Mean velocity gain (+/-SEM) for the horizontal component was 0.61 +/- 0.04 in darkness and increased to 0.72 +/- 0.03 for visible targets (P < 0.05), and for the vertical component was 0.54 +/- 0.02 in darkness, not significantly different from horizontal component gain. For visible targets, vertical component gain significantly increased to 0.63 +/- 0.04 (P < 0.05) with visible targets, but remained significantly less than horizontal component gain. PMID- 15827007 TI - Interaction between cortical control of vestibular function and spatial representation in man. AB - An asymmetry of the vestibulo-ocular reflex is observed after unilateral posterior cortical lesions in human. The co-occurrence of vestibular deficits and visuo-spatial disorders suggests a functional link between the representation and update of space and the integration of vestibular information in the occipito parieto-temporal cortex of the right hemisphere. PMID- 15827008 TI - Superior canal dehiscence is not due to cephalic displacement of the labyrinth. AB - Superior semicircular canal dehiscence syndrome may have a congenital basis. CT scans of 44 control and 21 dehiscent superior canals were examined to determine if dehiscent canals were more cephalically placed or vertically oriented than control canals. Results showed that neither was the case. Instead, the defect may be in the process of ossification above the superior canal. PMID- 15827009 TI - Saccadic analysis for early identification of neurological involvement in Gaucher disease. AB - The early identification of a neurological involvement in Gaucher's Disease (GD) is essential for a correct dosage of the enzymatic therapy. The saccadic eye movement characteristics of 15 patients affected by the non-neurological GD form show normal peak velocity values and some slight alterations of the velocity profiles present in more than 70% of the recorded eye movements in four cases; two of them later developed epilepsy. This fact supports the hypothesis that the presence of alterations in the velocity profile of the most part of saccadic movements is an index of initial neurological involvement. Further evaluations are necessary in order to understand if an increase of therapy dose should be used in all the subjects presenting such saccadic alterations. PMID- 15827010 TI - The specificity and sensitivity of uninhibited COR in labyrinthine-defective patients. AB - The Cervico-ocular reflex (COR), which depends on proprioceptive afferents from neck muscles to the vestibular nucleus, makes little contribution to the stabilization of gaze in humans. If labyrinthine function is lost because of disease, however, the COR may increase in gain and assume greater importance in generating compensatory eye rotations during natural head movements. We tested the COR, i.e., eye movement responses to rotation of the trunk about the earth fixed head (swinging test with fixed head STFH). If the vestibular function is intact, normally there should be no nystagmus. A total of 88 labyrinthine defective patients and 78 controls were studied. Of the 166 subjects evaluated, 88 presented some kind of alteration (either central or peripheral) and had STFH results showing "no inhibition" in 85 patients. Therefore, the STFH's sensibility is 96.6%. The remaining 3.4% were false negatives (i.e., pathology that could not be detected by the test). Of the 166 subjects evaluated, 78 had normal results and normal STFH. Therefore, STFH's specificity was 100%. We conclude that the study of the COR through STFH is a simple, useful, and clinically reliable test which should improve our knowledge of the interesting relationship between VOR and COR. PMID- 15827011 TI - Recurrent vertigo in extrinsic compression of the brain stem. AB - We present four patients who suffered from recurrent vertigo crises, which in most cases lasted hours or even days. In all of them the coincident finding was the presence of vascular compression of the brainstem (mainly vertebral dolichoectasia). Given the fact that the vertigo was always episodic and in that in the two cases where a treatment with carbamazepine could be used the result was the total suppression of the crises, we propose an ictal mechanism for them. Our findings differ from previous descriptions because: 1) the crises last longer, hours or even days, and 2) the involvement of the brainstem structures was extrinsic. PMID- 15827012 TI - Use of topiramate (topamax) in a subgroup of migraine-vertigo patients with auditory symptoms. AB - Five-three percent of the patients who suffer from migraine present severe incapacity and need rest in bed. If we add to this the incapacity produced by vertigo, then the quality of life of these patients is seriously affected. Migraine/Vertigo (MV) should be another criterion in the selection of preventive treatment even when other criteria are not fulfilled. Auditory symptoms may accompany MV. We treated 10 patients with Topiramate in an open trial, twice a day, with an average dose of 100 mg/day. The treatment period for these patients ranges between 6 and 16 months, with a mean of 9. As of today, all patients present no crisis. Regarding auditory symptoms, all the patients referred that they were stabilized. The effect began quickly, from the first month in most patients as it has been reported in other studies. PMID- 15827013 TI - Abnormal eye movements predict disability in MS: two-year follow-up. AB - We conducted a two-year follow-up study of 40 patients with MS in whom we had reported that abnormal eye movements (AEM) were associated with greater general disability. AEM patients (17/40) remained significantly (p < .001) more disabled (median EDSS of 7.0) than those with normal eye movements (median EDSS of 5.0). AEM and great disability were associated with abnormal MRI signals in brainstem or cerebellum, where disease may involve control circuits for eye movements as well as descending motor pathways. PMID- 15827014 TI - Saccade velocity as a surrogate disease marker in spinocerebellar ataxia type 2. AB - We measured in 82 spinocerebellar ataxia type 2 (SCA2) patients and in 80 controls maximal saccade velocity (MSV) and correlated it to polyglutamine expansion size and disease duration. MSV is strongly decreased in SCA2 patients and is influenced mostly by polyglutamine size. PMID- 15827015 TI - Effect of 4-aminopyridine on upbeat and downbeat nystagmus elucidates the mechanism of downbeat nystagmus. AB - The potassium channel blocker 4-aminopyridine (4-AP) restored vertical smooth pursuit and gaze holding in light in one patient with upbeat (UBN) and in one with downbeat nystagmus (DBN). Without a visible target, however, 4-AP had no effect on UBN, but DBN vanished. We hypothesize that this difference in the effects of 4-AP, which is known to increase the excitability of cerebellar Purkinje cells, can be attributed to the different lesion sites involved in UBN and DBN. PMID- 15827016 TI - Smooth pursuit in patients with downbeat nystagmus. AB - Smooth pursuit and gaze holding in light and darkness was investigated in 19 patients with downbeat nystagmus (DBN) and 17 control subjects. Vertical downward smooth pursuit was selectively impaired in patients, and gaze holding with a visible target was only slightly better than in darkness. The selective impairment of downward pursuit and the poor visual suppression of ocular drift in DBN support the involvement of the floccular lobe, which participates in gaze holding and contains gaze-velocity Purkinje cells with a downward on-direction for smooth pursuit. PMID- 15827017 TI - Cerebellar influence in oculomotor phase-transition behavior. AB - The cerebellum plays an important role in predicting and producing regularly timed motor responses. In the present report, we examined the performance of SCA6 cerebellar patients and normal controls in a simple saccade tracking task. Our results imply that the cerebellum plays a role in maintaining predictive motor behavior once it has been established. PMID- 15827018 TI - Familial positional downbeat nystagmus and cerebellar ataxia: clinical and pathologic findings. AB - A family with progressive cerebellar ataxia is reported. The earlier signs of cerebellar dysfunction was positional downbeat nystagmus (PDBN). An autopsy of one member with PDBN, who died early in the disease of unrelated causes, showed loss of Purkinje cells primarily in the nodulus. PMID- 15827019 TI - Predictive saccades to a regularly alternating target in cerebellar patients. AB - Predictive saccade tracking (targets stepping back and forth at frequencies of 1.25 = 1.0 Hz) and self-paced saccades between two stationary targets were measured in patients with cerebellar degeneration and in normal subjects. Cerebellar patients produced similar predictive latencies and generated the same percentage of predictive saccades as did normal subjects. Only the variability in the self-paced saccade paradigm was different between the groups. PMID- 15827020 TI - The origin of downbeat nystagmus: an asymmetry in the distribution of on directions of vertical gaze-velocity Purkinje cells. AB - Various hypotheses on the origin of cerebellar downbeat nystagmus (DBN) have been presented; the exact pathomechanism, however, is still not known. Based on previous anatomical and electrophysiological studies, we propose that an asymmetry in the distribution of on-directions of vertical gaze-velocity Purkinje cells leads to spontaneous upward ocular drift in cerebellar disease, and therefore, to DBN. Our hypothesis is supported by a computational model for vertical eye movements. PMID- 15827021 TI - Drift in ocular counterrolling during static head tilt. AB - A decreasing drift of the counter-rolled eye position (OCR) during head tilt was recently described. The underlying mechanism is not known. OCR in eleven healthy subjects was recorded (Search coil, Skalar) during a head tilt paradigm in two test conditions. The head was tilted with a velocity below (test 1) and above (test 2) detection threshold for the semicircular canals (SC) and held static for eight minutes. A significant drift of OCR was revealed in test 2 (P = .0006, ANOVA) and close to significant in test 1 (P = .07). No statistical difference was found between the two test conditions. The results suggest that the OCR drift was not caused by the SC complex merely. PMID- 15827022 TI - Retention of saccadic adaptation in humans. AB - In the present study, we tested in human subjects the persistence of the oculomotor changes resulting from saccadic adaptation up to 19 days after exposure to the double step target protocol. The main results indicate that the reduction of saccade gain related to the adaptation session (mean gain change of 5 subjects = 22 +/- 4.7%) was partially but significantly retained after 1 day and 5 days (mean amount of retention = 36 +/- 17% and 19.7 +/- 13.3%, respectively) but was no longer significant at day 11 and 19. Unexpectedly, gain changes were larger for leftward than for rightward saccades. No change in saccade dynamics was observed. These data suggest that in humans, adaptive mechanisms induce long lasting changes in visually-guided saccade amplitude, probably reflecting plastic changes in the brain. PMID- 15827023 TI - Effect of blinks on saccades before smooth-pursuit eye-movement initiation. AB - The effect of blinks on the saccade frequency before the onset of step ramp smooth pursuit was assessed in healthy controls. Blinks elicited before smooth pursuit onset suppressed saccades in contrast to a control and a gap paradigm. The source of blink associated suppression of saccades remains unclear but is probably not caused by visual signals or the omnipause neurons. PMID- 15827024 TI - Pontine lesions may cause selective deficits of "slow" vergence eye movements. AB - To address the role of pontine nuclei in vergence control, eye movements to ramp ('slow vergence') and step targets ('fast vergence') were recorded in two patients with unilateral mediolateral pontine infarctions and in ten healthy controls. 'Slow' vergence and conjugate smooth pursuit eye movements were impaired while 'fast' vergence was not. We conclude that like smooth pursuit signals, vergence signals are distributed in the pontine nuclei. PMID- 15827025 TI - Internuclear ophthalmoparesis in episodic ataxia type 2. AB - Two patients sharing a novel mutation of the CACNA1A gene for P/Q calcium channels showed significant slowing of adducting saccades compared with normal subjects or patients with cerebellar disease. Internuclear ophthalmoparesis (INO) was clinically evident in one. While these findings might be specific to this mutation, INO in our patients with episodic ataxia type 2 suggested involvement outside the cerebellum, either in the brain-stem internuclear pathway or at the neuromuscular junction. PMID- 15827026 TI - Video-based eye tracking: our experience with Advanced Stimuli Design for Eye Tracking software. AB - We present an independent, flexible, and easily programmable software program for generating a wide set of visual stimuli paradigms in eye-movement studies. The software, called ASTIDET (Advanced Stimuli Design for Eye Tracking), has been interfaced in real time with a high speed video-based eye tracking system in order to obtain a reliable measurement of saccades. Two saccadic paradigms have been tested (gap and memory guided tasks) in 10 normal subjects. The preliminary results confirm that ASTIDET is a user-friendly software and can be interfaced with a video-based eye-tracking device in order to obtain reliable measurement of saccades. PMID- 15827027 TI - Neural integrator function in murine CACNA1A mutants. AB - Time constants of gaze holding are shortened in rocker and tottering mice, two strains whose cerebellar dysfunction stems from genetic alterations of the P/Q calcium channel. The finding suggests that in mice as in primates, the cerebellum contributes to the function of the neural integrator. Studying CACNA1A mutants may elucidate how cerebellar signals support gaze holding. PMID- 15827028 TI - Single or combined oblique muscle surgery in acquired and congenital superior oblique palsy. AB - Oblique muscle surgery with weakening of the inferior oblique muscle alone or in combination with a tuck of the superior oblique tendon reduces in a dose dependent relationship the vertical deviation in patients with an acquired or congenital superior oblique palsy. Reduction of cyclotropia and the V-sign is less dose dependent. PMID- 15827029 TI - Binocular control of saccades in idiopathic Parkinson's disease. AB - We focused on the saccade disconjugate control in idiopathic Parkinson's disease patients. Our data showed that in IPD patients the saccade precision was differently impaired in the two eyes--namely, the disconjugate component was larger than in controls--more for the remembered than for the reflexive task. PMID- 15827030 TI - Studies of eccentric gaze stability: effects of pitch head position on horizontal gaze-holding in patients with cerebellar disease. AB - We studied the effects of change of head position in the pitch plane on the ability to hold horizontal eccentric gaze in five patients with cerebellar disorders. All patient showed a change in the time constant of horizontal centripetal drift when the head was pitched forward or back. This result suggests that otolithic inputs can influence the neural integrator for horizontal eye movements and indicates the value of testing horizontal gaze holding in different head positions. PMID- 15827031 TI - Catalysis, subcellular localization, expression and evolution of the targeting peptides degrading protease, AtPreP2. AB - We have previously identified a zinc metalloprotease involved in the degradation of mitochondrial and chloroplast targeting peptides, the presequence protease (PreP). In the Arabidopsis thaliana genomic database, there are two genes that correspond to the protease, the zinc metalloprotease (AAL90904) and the putative zinc metalloprotease (AAG13049). We have named the corresponding proteins AtPreP1 and AtPreP2, respectively. AtPreP1 and AtPreP2 show significant differences in their targeting peptides and the proteins are predicted to be localized in different compartments. AtPreP1 was shown to degrade both mitochondrial and chloroplast targeting peptides and to be dual targeted to both organelles using an ambiguous targeting peptide. Here, we have overexpressed, purified and characterized proteolytic and targeting properties of AtPreP2. AtPreP2 exhibits different proteolytic subsite specificity from AtPreP1 when used for degradation of organellar targeting peptides and their mutants. Interestingly, AtPreP2 precursor protein was also found to be dual targeted to both mitochondria and chloroplasts in a single and dual in vitro import system. Furthermore, targeting peptide of the AtPreP2 dually targeted green fluorescent protein (GFP) to both mitochondria and chloroplasts in tobacco protoplasts and leaves using an in vivo transient expression system. The targeting of both AtPreP1 and AtPreP2 proteases to chloroplasts in A. thaliana in vivo was confirmed via a shotgun mass spectrometric analysis of highly purified chloroplasts. Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that AtPreP1 and AtPreP2 are differentially expressed in mature A. thaliana plants. Phylogenetic evidence indicated that AtPreP1 and AtPreP2 are recent gene duplicates that may have diverged through subfunctionalization. PMID- 15827032 TI - Camurati-Engelmann disease--a case report and literature review. PMID- 15827033 TI - A defect in cortisol production in rheumatoid arthritis: why are we still looking? PMID- 15827034 TI - Ultraviolet-A1 phototherapy modulates Th1/Th2 and Tc1/Tc2 balance in patients with systemic lupus erythematosus. AB - OBJECTIVE: Ultraviolet-A1 (UVA1) phototherapy is effective for a variety of dermatological diseases. We examined the effectiveness and reliability of low dose UVA1 phototherapy (60 kJ/m2/treatment) in patients suffering from systemic lupus erythematosus (SLE). We studied the changes in immunological parameters. METHODS: The patients received a 9-week course of phototherapy according to the following regimen: five times a week during the first 3 weeks, three times a week during the second 3 weeks and twice during the last 3 weeks. Among other things, we analysed the proportions of T helper 1 (Th1), Th2, T cytotoxic (Tc1) and Tc2 cell populations in the peripheral blood of patients by flow cytometric detection of intracytoplasmic interferon gamma (IFN-gamma) and interleukin 4 (IL-4). RESULTS: Our study showed the improvement of clinical symptoms determined by the subjective clinical disease activity scoring and the SLE Disease Activity Index (SLEDAI). By the end of UVA1 phototherapy, the mean value of SLEDAI had decreased from 7.2+/-5.6 to 0.9+/-1.8, which was significant (P = 0.005). Immunological investigations detected a decrease in the frequency of IFN-gamma-producing Th1 and Tc1 cells and a decrease in the Th1/Th2 and Tc1/Tc2 ratios after UVA1 therapy. CONCLUSION: According to the literature, IFN-gamma has a pathogenic role in the development of SLE. We observed a decreased proportion of IFN-gamma secreting cells, which we think is presumably one of the beneficial effects of UVA1 therapy. On the basis of our study, UVA1 phototherapy does seem to be an effective adjuvant in the treatment of SLE patients. PMID- 15827035 TI - A regional audit of the use of COX-2 selective non-steroidal anti-inflammatory drugs (NSAIDs) in rheumatology clinics in the West Midlands, in relation to NICE guidelines. AB - OBJECTIVES: Whilst all non-steroidal anti-inflammatory drugs (NSAIDs) can cause adverse gastrointestinal events, COX-2-selective inhibitors (COX-2) may have improved gastrointestinal safety compared with non-selective NSAIDs (NSNSAIDs). In 2001, the National Institute for Clinical Excellence (NICE) published guidance on the use of the COX-2 agents celecoxib, rofecoxib, meloxicam and etodolac for rheumatoid arthritis (RA) and osteoarthritis (OA). This study aimed to audit the appropriateness of NSAID use in relation to NICE guidance in rheumatology out patients. METHODS: Questionnaires were completed for all patients attending clinics in 18 rheumatology units in the West Midlands over a 2-week period. Data collected included patient demographics, NSAID type, indications, duration of use (> or =3 months was considered prolonged), and concomitant prescription of corticosteroids, warfarin and gastroprotective agents. RESULTS: Data were collected on 2846 patients; 1164 (41%) were taking NSAIDs (791 NSNSAIDs, 373 COX 2). Of the 1164 NSAID users, 753 (65%) had a diagnosis of RA or OA (483 NSNSAIDs, 270 COX-2). Overall, 37% of NSAID prescriptions were appropriate. Of the NSNSAID users, 92% had at least one risk factor for adverse gastrointestinal events and were therefore inappropriately treated. Prolonged use (in 89%) and age > or =65 yr (in 23%) were the most frequent risk factors identified. Of the COX-2 users, 97% had one or more risk factors and were appropriately treated. Analysis of the RA/OA subgroup revealed similar findings. Thirty-six per cent were taking NSAIDs appropriately; 97% of NSNSAID use was inappropriate and 97% of COX-2 use was appropriate treatment. In the whole cohort, gastroprotective agents were used in 26% of NSNSAID users, 56% of gastroprotective agents being proton pump inhibitors. CONCLUSIONS: Ninety-two per cent of patients attending rheumatology clinics who were taking NSNSAIDs should have been prescribed a COX-2-selective agent in relation to NICE guidance. Duration of use and age > or =65 yr emerged numerically as the most important risk factors. Significant numbers of patients taking NSNSAIDs may be at risk from adverse gastrointestinal events and clinicians may wish to review their prescribing patterns. Conversely, 97% of patients taking COX-2 agents were treated appropriately. Although practice overall conformed poorly with NICE guidance, NSAID prescribing also needs to be considered in the context of recent concerns regarding the cardiovascular risks of COX-2 agents. PMID- 15827036 TI - An exacerbation of Behcet's syndrome presenting with bilateral papillitis. PMID- 15827037 TI - Joint hypermobility in children. PMID- 15827038 TI - Platelet-endothelial cell interactions in murine antigen-induced arthritis. AB - OBJECTIVES: Growing evidence supports the substantial pathophysiological impact of platelets on the development of rheumatoid arthritis. At present there are no methods for studying these cellular mechanisms in vivo. The aim of this study was to visualize and investigate platelet-endothelial cell interaction in the knee joint of mice with antigen-induced arthritis (AiA) by means of intravital microscopy. METHODS: In 14 mice (Balbc) intravital microscopic assessment was performed on day 8 after AiA induction in two groups (controls, AiA). The severity of AiA was assessed by measuring knee joint swelling and by histological scoring. Ex vivo fluorescently labelled rolling and adherent platelets and leucocyte-endothelium interactions were investigated by intravital fluorescence microscopy. RESULTS: Swelling of the knee joint as well as histological score was significantly enhanced in arthritic animals compared with controls. In control mice intravital microscopy revealed low baseline rolling and sticking of leucocytes and fluorescently labelled platelets. AiA induced a significant increase in the fraction of rolling leucocytes (3 times) and rolling platelets (6 times) compared to the control group. Furthermore, AiA induction resulted in a significantly enhanced number of adherent leucocytes (3-fold) and adherent platelets (12-fold) in comparison with control animals. CONCLUSIONS: Platelet kinetics were directly analysed using intravital microscopy in the arthritic microcirculation in vivo for the first time. We provide the first evidence that platelets accumulate in arthritic vessels, indicating platelet activation due to AiA. Platelet recruitment and subsequent activation might play an important role in the pathogenesis of rheumatoid arthritis. PMID- 15827039 TI - Arthritis in association with human immunodeficiency virus infection in Black African children: causal or coincidental? AB - OBJECTIVES: To compare human immunodeficiency virus (HIV)-infected and HIV uninfected children with arthritis of unknown origin to determine whether the association between HIV infection and arthritis is causal or coincidental. METHOD: Retrospective review of 132 children with arthritis who were tested for HIV infection. RESULTS: Thirty-five (27%) of the children were HIV infected and the male to female ratio was 2.5:1 (P = 0.02). Arthritis was the presenting feature of HIV infection in 78% of these children. The remaining 97 (73%) were diagnosed as having juvenile idiopathic arthritis. 'Spondyloarthropathy-like' features were found in 34% of HIV-infected children compared with 5% of uninfected children. CONCLUSION: The high prevalence of HIV infection in 27% of children, the predominance of males and the increased prevalence of 'spondyloarthropathy-like' features, supports a causal relationship between HIV infection and arthritis. PMID- 15827040 TI - Necrotizing scleritis associated with rheumatoid arthritis: long-term remission with high-dose infliximab therapy. PMID- 15827041 TI - Insufficiency fracture of the tarsal navicular in a patient with rheumatoid arthritis. PMID- 15827042 TI - Anti-endothelial cell antibodies from lupus patients bind to apoptotic endothelial cells promoting macrophage phagocytosis but do not induce apoptosis. AB - OBJECTIVE: Anti-endothelial cell antibodies (AECA) have been reported to induce apoptosis. We investigated the induction of apoptosis by these autoantibodies and their involvement in the removal of apoptotic cells. METHODS: AECA isolated from patients with active systemic lupus erythematosus (SLE) were incubated with human umbilical vein endothelial cells (HUVECs). AECA-positive sera were identified using a cell-based ELISA. Apoptosis was measured by morphology and phosphatidylserine externalization using flow cytometry with fluorescein isothiocyanate (FITC)-conjugated annexin V. Flow cytometry was used to investigate AECA binding to apoptotic cells using FITC-conjugated anti-human immunoglobulin G (IgG). Apoptotic endothelial cells were stained with a red dye (PKH26) and co-cultured with macrophages, and phagocytosis was visualized under phase contrast microscopy. RESULTS: AECA from patients with SLE did not induce apoptosis compared with normal IgG (nIgG) at any time point, as assessed by morphology (at 24 h, P = 0.167) or phosphatidylserine externalization (at 24 h, P = 0.098). However, there was increased binding of AECA to apoptotic endothelial cells (48.8 +/- 11.9 compared with 25.8 +/- 6.7% AECA binding to freshly isolated cells, P< 0.001). These opsonized endothelial cells showed greater phagocytosis by macrophages (mean phagocytic index 24.9 +/- 4.5%) when cells opsonized with nIgG were compared with AECA (34.8 +/- 3.4% n = 5, P = 0.01). CONCLUSION: In conclusion, AECA bind to apoptotic endothelial cells but do not induce endothelial cell apoptosis. Macrophage phagocytosis is increased by opsonization of apoptotic endothelial cells by AECA, a proinflammatory mechanism of cell removal. PMID- 15827043 TI - Sensory-motor incongruence and reports of 'pain'. PMID- 15827044 TI - Outcome following onset of juvenile idiopathic inflammatory arthritis: II. predictors of outcome in juvenile arthritis. AB - OBJECTIVE: To assess the relative contributions of demographic, clinical and laboratory variables in predicting outcome in juvenile idiopathic inflammatory arthritis (JIA), based on a review of the existing literature. METHODS: Electronic reference database searches for the previous 10 yr were conducted and studies examining the role of major potential predictors of main outcomes were identified. Where possible, subjects were grouped by JIA disease subtype. In addition to demographic variables, the following disease-related predictors were assessed: nature of joint involvement, acute-phase response, and presence of autoantibodies. These were then analysed for three main outcomes of interest: remission as assessed by disease activity; functional impairment; and structural damage as assessed by radiological joint erosions. RESULTS: In general, female gender, polyarticular and symmetrical joint involvement, elevated inflammatory markers and rheumatoid factor positivity were the most consistent predictors of a poor outcome, although the studies were frequently inconsistent in both the direction and the magnitude of the effects. CONCLUSIONS: These data are too variable to accurately identify those predictors associated with poor outcome following the onset of JIA. Although some of this variation may be the result of true differences between study populations, the vast majority of inconsistencies are explainable by the absence of standardized classification systems, outcome definitions, therapeutic approach and research tools. More comprehensive prospective evaluation is required before robust prediction models can be generated. PMID- 15827045 TI - Outcome following onset of juvenile idiopathic inflammatory arthritis: I. frequency of different outcomes. AB - OBJECTIVE: To determine the outcome, following the onset of juvenile idiopathic inflammatory arthritis, in terms of remission of disease activity, loss of function and structural damage based on a review of the available published data. METHODS: Electronic databases were searched for major studies publishing outcome data in the past 10 yr in juvenile idiopathic arthritis, juvenile rheumatoid arthritis and juvenile chronic arthritis, and 21 studies were selected. The proportions of children in the different categories of the outcomes of interest are described. Data were stratified where possible by disease subtype. RESULTS: There were major differences between the studies reviewed in terms of study design, case selection and the results obtained. In general, children with systemic- or polyarticular-onset disease were much less likely to go into remission than those with oligoarticular onset, although the remission rates in the latter group ranged from 36 to 84%. Several different approaches were used to assess functional outcome but the pattern of results between the different subgroups was the same as with remission. Similarly, children with polyarticular disease in all the cohorts reviewed were substantially more likely to have erosive radiological damage on follow-up. The rates of individual outcomes, even within a subgroup, varied considerably between studies and this does not appear to be explained solely by differences in methodology. CONCLUSIONS: There remains a considerable lack of clarity in the prognosis following onset of juvenile idiopathic arthritis for the major outcomes considered, although those with oligoarthritis at presentation have the best outcome. The ability to offer accurate prognosis is particularly important to both reassure parents and guide treatment at disease onset. To achieve this, large definitive prospective studies will be required. PMID- 15827046 TI - Renal toxicity of oxaliplatin. PMID- 15827047 TI - Reduced baroreflex sensitivity is associated with increased vascular calcification and arterial stiffness. AB - INTRODUCTION: Vascular calcification is a critical determinant of cardiovascular morbidity and mortality in chronic haemodialysis (HD) patients. The pathophysiology underlying this observation remains obscure. Baroreceptor sensitivity (BRS) is important in the maintenance of an appropriate cardiovascular status both at rest and under the physiological stress of HD. BRS is determined by both the mechanical properties of the vascular wall, mediating the transfer of transmural pressure, and afferent and efferent autonomic function. We aimed to study the association between arterial structure, function and BRS in chronic HD patients. METHODS: We studied 40 chronic HD patients mean age 62+/-2 (26-86) years who had received HD for a mean 40+/-4 (9-101) months. Spontaneous BRS was assessed using software studying the relationship between inter-beat variability and beat to beat changes in systolic blood pressure. Functional characteristics of conduit arteries (pulse wave analysis) were studied with applanation tonometry at the radial artery. Arterial calcification was assessed in lower limbs using reconstructed multi-slice computed tomography and quantified with volume-corrected calcification scores within the superficial femoral artery. RESULTS: Mean BRS was 4.43+/-0.44 ms/mmHg, with a wide range from 1.0 to 11.5 ms/mmHg. This correlated with arterial stiffness as measured by time to shoulder calculated from the central pulse wave analysis (r = 0.4, P = 0.01). BRS was also associated with vascular calcification (P = 0.01) but not by other factors such as dialysis vintage, age or pre-dialysis systolic/diastolic blood pressure. CONCLUSION: The reduction in BRS and the resulting aberrant blood pressure response to the physiological stress and volume changes of HD may be important in the further understanding of the pathophysiology of the increased mortality in HD patients with vascular calcification. PMID- 15827048 TI - A detailed analysis of sodium removal by peritoneal dialysis: comparison with predictions from the three-pore model of membrane function. AB - BACKGROUND: The development of fluid and salt retention is a potential problem for all peritoneal dialysis (PD) patients. Sodium removal by the peritoneum is predominantly determined by convective fluid loss but influenced by diffusion and sieving due to free water transport as predicted by the three-pore model (TPM). The aim of the study was to establish the effect of transport status, dwell length and glucose concentration on observed ultrafiltration (UF), dialysate sodium concentration ([Na(+)](D)) and removal, and compare this with that predicted by a computer program based on the principles of the TPM. METHODS: This was a cross-sectional study of UF and [Na(+)](D) collected prospectively from dwells classified by length, glucose concentration and membrane transport characteristics. Solute transport, converted to area parameter and UF capacity, was measured on each occasion by the peritoneal equilibration test. These parameters, along with plasma [Na(+)], were entered into the computer model. Fixed values for other parameters, e.g. hydraulic conductance and lymphatic absorption and sump volume, were used. RESULTS: A total of 1853 dwells from 182 patients [10% were on automated PD (APD)] were analysed. There was a high degree of correlation (r = 0.83-95, P<0.001) between the observed and predicted values for UF, [Na(+)](D) and sodium removal across the full range of dwell categories. The model overpredicted UF as the net volume increased with increasing glucose concentration, independently of solute transport. This bias was not fully explained by the preferential use of hypertonic dialysate by patients with reduced UF capacity. The prediction of [Na(+)](D) described sodium sieving, which was overestimated in a small number of patients with UF failure. There were no discrepancies between continous ambulatory PD (CAPD) and APD patients. CONCLUSION: This analysis endorses the TPM as a description of membrane function, particularly in relation to sodium sieving and removal. The relationship between dialysate glucose concentration and achieved UF appears to be more complex; even accounting for extended time on treatment and reduction in the osmotic conductance in patients preferentially using hypertonic exchanges, further adjustments may be needed to account for the tendency to overestimate UF. PMID- 15827049 TI - Disturbed calcium metabolism in a patient with bipolar disorder and impaired renal function. PMID- 15827050 TI - Factors influencing peritoneal transport parameters during the first year on peritoneal dialysis: peritonitis is the main factor. AB - BACKGROUND: Studies on the evolution of peritoneal transport during the first year of peritoneal dialysis (PD) are scarce and their results are contradictory. The aim of the present study was to analyse the evolution of peritoneal transport and residual renal function during the first year on PD, and to determine the factors that may influence them. METHODS: We studied 249 patients on continuous ambulatory PD with glucose exchange solutions (117 men, 132 women, mean age 51.9+/-16 years) 59 of whom had diabetes (25 type I). At baseline and after 1 year, we determined the mass transfer coefficients of urea (U-MTAC) and creatinine (Cr-MTAC), net ultrafiltration and residual renal function. RESULTS: Residual renal function decreased significantly during the first year (from 3.9+/ 2.8 to 2.4+/-2.2 ml/min, P<0.001). Both U-MTAC and Cr-MTAC decreased after 1 year [U-MTAC from 22.7+/-7.8 to 20.7+/-6.6 ml/min (P<0.001), Cr-MTAC from 10.5+/-5.3 to 10.1+/-4.6 ml/min (NS)]. The ultrafiltration capacity increased significantly (from 923+/-359 to 987 U 341 ml/4 h, P<0.001). The evolution of MTAC values was independent of age, sex, diabetes and amount of hypertonic glucose used. When patients were grouped according to their initial Cr-MTAC, we observed a tendency toward normalization of the parameters of peritoneal function. Patients with peritonitis (n = 88) showed a first year increase in Cr-MTAC, which was significantly higher than in patients without peritonitis (11.1+/-5 vs 9.5+/-4.2, P<0.01). Ultrafiltration decreased in patients with more than four accumulated days of peritonitis (from 1062+/-447 to 1024+/-340 ml/4 h, NS); it increased in patients without peritonitis. CONCLUSIONS: The peritoneal transport parameters tended toward normalization during the first year on PD, mainly with a decrease of small solute transport and an increase of ultrafiltration capacity. This evolution is independent of age, gender, diabetes and higher exposure to glucose in PD solutions. Peritonitis was the only independent factor that affected peritoneal function during the first year on peritoneal dialysis. PMID- 15827051 TI - Prolonged treatment of refractory Wegener's granulomatosis with 15 deoxyspergualin: an open study in seven patients. AB - BACKGROUND: A subset of patients with Wegener's granulomatosis does not respond to daily oral cyclophosphamide (CYC) plus corticosteroids or suffers from intolerable side effects. A 6 month course of the immunosuppressant 15 deoxyspergualin (DSG) has previously been employed successfully in these refractory cases. However, there are no reports on long-term treatment with DSG. METHODS: To document the effects of prolonged DSG treatment, this study reports on seven patients suffering refractory Wegener's granulomatosis, who were successfully treated with DSG over an average of 26.5 months (range: 11-55.5 months). RESULTS: Before administration of DSG, patients had experienced an average of 6.6 relapses (range: 3-12) under an average of 5.4 (range: 2-11) different therapeutic approaches, which included CYC in all cases. All suffered active disease when DSG was initiated. Four were unresponsive to CYC and three did not tolerate it. DSG (0.5 mg/kg/day subcutaneous) was given for 2-3 weeks until the leukocyte count dropped to 3000/microl, followed by a rest until a leukocyte count of 4000/microl was reached again. No other immunosuppressants besides corticosteroids were given. All patients showed a long-lasting, favourable response to DSG with complete (n = 5) or partial (n = 2) remission. Only one case relapsed while being treated with DSG. Termination/interruption of DSG was followed by relapse in four of five occasions. Resumption of DSG led to complete remission. Currently, five of the seven patients are still treated with DSG and are in remission. Infections, mainly of the respiratory tract, were observed in five cases and resolved after treatment. One case developed a third degree heart block that required pacing. CONCLUSIONS: In patients with refractory Wegener's granulomatosis, prolonged treatment with DSG seems safe and successful. PMID- 15827052 TI - Foreword. PMID- 15827053 TI - Assays for detecting and diagnosing antibody-mediated pure red cell aplasia (PRCA): an assessment of available procedures. AB - Antibody (Ab)-mediated pure red cell aplasia (PRCA) develops when patients mount a neutralizing Ab response to recombinant erythropoiesis-stimulating agents (ESAs) such as epoetin-alpha (EPO). These neutralizing Abs can also cross neutralize endogenous EPO, leading to a state of absolute EPO resistance and transfusion dependence. The diagnosis of Ab-mediated PRCA in part relies on the sensitive and specific detection of serum anti-EPO Abs and a confirmatory examination of patients' bone marrow for lack of erythroid precursors. To date, a variety of assays have been used to detect anti-EPO Abs, including radioimmunoprecipitation (RIP) assay, enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR) and bioassays that measure neutralizing Abs. Each of these assays can yield informative results and possess characteristic benefits and limitations, so it is unclear whether or not a "superior" assay is required or possible. To date, a universal standardized assay has not yet been established that would facilitate the comparison of Ab data derived from different laboratories and retrospective analysis of stored sera. This review evaluates the results of studies measuring anti-EPO Abs with different assays and compares their relative advantages and disadvantages in terms of specificity, sensitivity, ease of use and ability to measure Ab-binding affinities and subclasses. These comparisons provide a basis for determining the optimal assay(s) for screening and/or analysis of patients' serum for anti-EPO Abs during treatment or after onset of Ab-mediated PRCA. PMID- 15827054 TI - Antibody-mediated pure red cell aplasia (PRCA) treatment and re-treatment: multiple options. AB - In the vast majority of patients with antibody (Ab)-mediated pure red cell aplasia (PRCA), simple withdrawal of the erythropoiesis-stimulating agent (ESA) does not effectively reverse PRCA. In contrast, immunosuppressive treatments can induce the disappearance of anti-erythropoietin Abs and a reversal of PRCA. Consensus opinion on the optimal therapy has not been established, but individual case reports or case series suggest that kidney transplantation or treatment with corticosteroids plus cyclophosphamide are the most effective therapies. However, treatment with cyclosporine is an interesting alternative, since it appears to be effective in at least two-thirds of patients and with minimal side effects. Due to the key role of ESAs in the management of patients with chronic kidney disease (CKD), some patients have been re-treated with an ESA following resolution of Ab mediated PRCA. In all reported cases, this treatment increased haemoglobin levels, alleviated the need for transfusions and did not have side effects. However, one should be extremely cautious when deciding to re-treat a patient with ESA, due to the small number of reported cases and the possibility of publication bias. PMID- 15827055 TI - Antibody-mediated pure red cell aplasia (PRCA): ensuring future progress by collecting data from a registry. AB - The recent emergence of antibody (Ab)-mediated pure red cell aplasia (PRCA) resulting from administration of certain erythropoiesis-stimulating agents (ESAs) has heightened awareness of the potential for this disorder in patients receiving ESAs for anaemia associated with chronic kidney disease (CKD). The Spanish Society of Nephrology sponsored an independent registry for analysis of patients who developed epoetin-induced, Ab-mediated PRCA in Spain. Twelve patients from 11 regional hospitals were included in the Spanish PRCA registry from November 2000 to December 2002 that met the criteria for Ab-mediated PRCA. Patients were reported using a standardized form specifically developed for PRCA, and serum samples were analysed in a central laboratory at Reina Sofia University Hospital in Cordoba, Spain. The characteristics of these patients, their serological and haematological results, and the outcomes of immunosuppressive therapies are presented. The Spanish PRCA registry serves as a model for establishing an independent global PRCA registry sponsored by the various nephrology societies in European Union countries, and elsewhere and coordinated by key investigators from the respective countries. PMID- 15827056 TI - What is antibody-mediated pure red cell aplasia (PRCA)? AB - Antibody (Ab)-mediated pure red cell aplasia (PRCA) is an immunological pathology associated with the production of neutralizing Abs that inhibit the erythropoietic activity of endogenous erythropoietin (EPO) and recombinant erythropoiesis-stimulating agents (ESAs). Although this disorder occurs very rarely, the number of reported cases has increased dramatically in recent years, predominantly in patients with chronic kidney disease (CKD)-associated anaemia receiving subcutaneous (s.c.) injections of one particular formulation of recombinant epoetin-alpha. This disorder is differentiated from classic forms of PRCA that are caused by chemical toxaemia (i.e. erythroblastopenia induced by chemical compounds), lymphoproliferative neoplasms, thymoma, human parvovirus B19 and certain autoimmune disorders. Patients with Ab-mediated PRCA develop resistance to EPO and severe anaemia that follows a period of successful erythropoietic response, and exhibit characteristic decreases in blood haemoglobin (Hb) level and in the number of circulating reticulocytes. However, it is not yet possible to predict which patients will develop PRCA or when in the course of their treatments PRCA may develop. Laboratory confirmation of Ab mediated PRCA requires bone marrow examination demonstrating few or no erythroid precursors and the presence of serum anti-EPO Abs using a validated assay. These neutralizing anti-EPO Abs recognize the protein core of the EPO molecule; carbohydrate groups on EPO can affect the binding of Abs but are themselves not immunological determinants. Animal models are being developed to increase further our understanding of the immunological mechanisms underlying the onset and progression of Ab-mediated PRCA. PMID- 15827057 TI - Follow-on biologics: challenges of the "next generation". AB - The imminent patent expiration of many biopharmaceutical products will produce the possibility for generic versions of these therapeutic agents (i.e. biosimilars). However, there are a number of issues that will make approval of biosimilars much more complicated than the approval of generic equivalents of conventional pharmaceuticals. These issues centre on the intrinsic complexity of biopharmaceutical agents, which are recombinant proteins in most cases, and the heterogeneity of proteins produced by different manufacturing processes (i.e. differences in host cells, purification and processing, formulation and packaging). The increased occurrence of antibody (Ab)-mediated pure red cell aplasia (PRCA) associated with a change in the formulation of one particular epoetin-alpha product highlights the potential for increased immunogenicity of recombinant proteins with different formulations, or those manufactured by different processes. Thus, verification of the similarity to or substitutability of biosimilars with reference innovator biopharmaceutical products will require much more than a demonstration of pharmacokinetic similarity, which is sufficient for conventional, small molecule generic agents. Regulatory requirements for the approval of biosimilars have not yet been fully established, but preliminary guidelines from the European Agency for the Evaluation of Medicinal Products (EMEA) state that the complexity of the product, the types of changes in the manufacturing process, and differences in quality, safety and efficacy must be taken into account when evaluating biosimilars. For most products, results of clinical trials demonstrating safety and efficacy are likely to be required. In addition, because of the unpredictability of the onset and incidence of immunogenicity, extended post-marketing surveillance is also important and may be required. PMID- 15827058 TI - Antibody-mediated pure red cell aplasia (PRCA): epidemiology, immunogenicity and risks. AB - Although epoetin-induced antibody (Ab)-mediated pure red cell aplasia (PRCA) was very rare prior to 1998, a large increase in the number of global cases was observed from 1999 to 2002 in patients treated with erythropoiesis-stimulating agents (ESAs) for the anaemia associated with chronic kidney disease (CKD). The number of global cases of this immunological form of PRCA has declined precipitously since 2003 following increased awareness of this disorder and changes in the handling and administration of the formulation of epoetin-alpha (EPO) that was associated with the majority of cases. Current recommendations state that patients should stop treatment immediately following a diagnosis of Ab mediated PRCA and not resume treatment with the same or another ESA. The feasibility of re-treatment following remission from PRCA or during ongoing immunosuppressive therapy is currently under investigation. The immunological mechanism for developing Ab-mediated PRCA is unknown, but a variety of factors may increase the immunogenicity of epoetin or other ESAs. Product-related factors that have the potential to impact on immunogenicity include sequence variations in proteins, the degree and nature of protein glycosylation, the manufacturing process, handling and storage, and components and properties of the product formulation. Patient-related factors associated with developing Ab-mediated PRCA include skin reactions, immune status and treatment history. Increased awareness among physicians of the factors contributing to the development of PRCA and its distinguishing clinical features has coincided with studies aimed at identifying effective therapies for this disorder. A number of immunosuppressive therapies have been shown recently to reconstitute effectively the erythropoietic response in patients with PRCA. However, therapeutic approaches for this serious immunological reaction to recombinant ESAs remain investigative. PMID- 15827059 TI - Arterial stiffness: intermediate or surrogate endpoint for cardiovascular events? PMID- 15827060 TI - Personality traits and heart rate variability predict long-term cardiac mortality after myocardial infarction. AB - AIMS: To investigate personality traits and sympatho-vagal modulation of heart rate variability (HRV) during acute myocardial infarction (AMI), assessing their relationships and their long-term prognostic value. METHODS AND RESULTS: Psychological traits and 24 h HRV were prospectively investigated in 246 patients at discharge of an AMI. Patients were followed-up to 8 years for the occurrence of cardiac death and non-fatal reinfarction. Low coping and anxiety traits associated with reduced HRV characterized the study population. At univariate analysis, low emotional sensitivity and insecurity, relative tachycardia, reduced high frequency (HF), and low frequency power and pNN50 were predictive of cardiac death at 8-year follow-up. At multivariable analysis, low emotional sensitivity and low HF power remained predictive, with a relative risk of 4.18 (P=0.003) and 2.76 (P=0.007), respectively; also the type of infarction (Q vs. non-Q) and hospital length of stay were independent predictive variables. CONCLUSION: Anxiety and emotional sensitivity were significant predictors of 8-year cardiac mortality after AMI. Reduced HF power, a recognized marker of vagal withdrawal, increased the risk. PMID- 15827061 TI - Adherence to guidelines is a predictor of outcome in chronic heart failure: the MAHLER survey. AB - AIMS: The impact on outcome of the implementation of European guidelines for the treatment of chronic heart failure (CHF) has not been evaluated. We investigated the consequences of adherence to care by cardiologists on the rate of CHF and cardiovascular (CV) hospitalizations and time to CV hospitalization. METHODS AND RESULTS: We constructed class adherence indicators for angiotensin-converting enzyme (ACE)-inhibitors, beta-blockers, spironolactone, diuretics, and cardiac glycosides and GAIs (GAI3 adherence to first three classes of heart failure medication, GAI5 adherence to five classes). In the study, 1410 evaluable patients (mean age 69, 69% males, New York Heart Association (NYHA) II: 64%, III: 34%, IV: 2%) were enrolled and followed up for 6 months by 150 randomly selected cardiologists/cardiology departments from six European countries (France, Germany, Italy, The Netherlands, Spain, and UK). Overall, adherence to treatment guidelines was 60 (GAI3) and 63% (GAI5) and was better for ACE-I (88%) or diuretics (82%) than for cardiac glycosides (52%), beta-blockers (58%), and spironolactone (36%). In the three tertiles of the population defined by a decreasing mean adherence score value, CHF and CV hospitalization rates were, respectively, 6.7, 9.7, and 14.7% and 11.2, 15.9, and 20.6% (P<0.002 and P<0.001, respectively). Global adherence indicator GAI3 was an independent predictor of time to CV hospitalization in a multi-variable model together with NYHA Class, history of CHF hospitalization, ischaemic aetiology, diabetes mellitus, and hypertension. CONCLUSION: We demonstrate that adherence of physicians to treatment guidelines is a strong predictor of fewer CV hospitalizations in actual practice. There is a need to develop further quality improvement programmes in this condition. PMID- 15827062 TI - Extracellular matrix remodelling in human aortic valve disease: the role of matrix metalloproteinases and their tissue inhibitors. AB - AIMS: Aortic valve diseases are characterized by pathological remodelling of valvular tissue but the cellular and molecular effectors involved in these processes are not well known. The role of matrix metalloproteinase (MMP)-2, MMP 9, MMP-3, MMP-7, and tissue inhibitor of matrix metalloproteinase (TIMP)-1 and TIMP-2 are investigated here. METHODS AND RESULTS: Histological analysis of pathological valves [aortic stenosis (AS) (n=49), aortic regurgitation (AR) (n=23)] and control valves (n=8) was performed. The main tissue abnormalities (calcification, inflammatory cells, and capillaries) observed in AS were less severe or absent in AR. However, both groups of pathological valves displayed similar histological signs of extracellular matrix (ECM) remodelling. Biochemical analysis of MMPs and TIMPs (gelatin and casein zymography and ELISA) was performed on valve extracts. MMP-2 activity was not significantly different in control and pathological valves. Increases in MMP-9 and MMP-3 in AS demonstrated an inflammatory state. Finally, there was a four- to seven-fold increase of TIMP 1 in pathological valves. TIMP-1, TIMP-2, and MMP-2 were synthesized by the valvular interstitial cells in primary culture. CONCLUSION: This study demonstrates the involvement of the MMP/TIMP system in ECM remodelling of both AS and AR. These findings provide evidence of inflammatory injury more severe in AS than in AR and involvement of mesenchymal cell response. PMID- 15827063 TI - Open access and openly accessible: a study of scientific publications shared via the internet. AB - OBJECTIVES: To determine how often reprints of scientific publications are shared online, whether journal readership level is a predictor, how the amount of file sharing changes with the age of the article, and to what degree open access publications are shared on non-journal websites. DESIGN: The internet was searched using an application programming interface to Google, a popular and freely available search engine. MAIN OUTCOME MEASURES: The proportion of reprints of journal articles published between 1994 and 2004 from within 13 subscription based and four open access journals that could be located online at non-journal websites. RESULTS: The probability that an article could be found online at a non journal website correlated with the journal impact factor and the time since initial publication. Papers from higher impact journals and more recent articles were more likely to be located. On average, for the high impact journal articles published in 2003, over a third could be located at non-journal websites. Similar trends were observed for the delayed or full open access publications. CONCLUSIONS: Decentralised sharing of scientific reprints through the internet creates a degree of de facto open access that, though highly incomplete in its coverage, is none the less biased towards publications of higher popular demand. PMID- 15827064 TI - Indinavir alters regulators of protein anabolism and catabolism in skeletal muscle. AB - The HIV protease inhibitor indinavir adversely impairs carbohydrate and lipid metabolism, whereas its influence on protein metabolism under in vivo conditions remains unknown. The present study tested the hypothesis that indinavir also decreases basal protein synthesis and impairs the anabolic response to insulin in skeletal muscle. Indinavir was infused intravenously for 4 h into conscious rats, at which time the homeostasis model assessment of insulin resistance was increased. Indinavir decreased muscle protein synthesis by 30%, and this reduction was due to impaired translational efficiency. To identify potential mechanisms responsible for regulating mRNA translation, several eukaryotic initiation factors (eIFs) were examined. Under basal fasted conditions, there was a redistribution of eIF4E from the active eIF4E.eIF4G complex to the inactive eIF4E.4E-BP1 complex, and this change was associated with a marked decrease in the phosphorylation of 4E-BP1 in muscle. Likewise, indinavir decreased constitutive phosphorylation of eIF4G and mTOR in muscle, but not S6K1 or the ribosomal protein S6. In contrast, the ability of a maximally stimulating dose of insulin to increase the phosphorylation of PKB, 4E-BP1, S6K1, or mTOR was not altered 20 min after intravenous injection. Indinavir increased mRNA expression of the ubiquitin ligase MuRF1, but the plasma concentration of 3-methylhistidine remained unaltered. These indinavir-induced changes were associated with a marked reduction in the plasma testosterone concentration but were independent of changes in plasma levels of IGF-I, corticosterone, TNF-alpha, or IL-6. In conclusion, indinavir acutely impairs basal protein synthesis and translation initiation in skeletal muscle but, in contrast to muscle glucose uptake, does not impair insulin-stimulated signaling of protein synthetic pathways. PMID- 15827065 TI - Proteomic analysis on insulin signaling in human hematopoietic cells: identification of CLIC1 and SRp20 as novel downstream effectors of insulin. AB - Insulin/IGF-I-dependent signals play important roles for the regulation of proliferation, differentiation, metabolism, and autophagy in various cells, including hematopoietic cells. Although the early protein kinase activation cascade has been intensively studied, the whole picture of intracellular signaling events has not yet been clarified. To identify novel downstream effectors of insulin-dependent signals in relatively early phases, we performed high-resolution two-dimensional electrophoresis (2-DE)-based proteomic analysis using human hematopoietic cells 1 h after insulin stimulation. We identified SRp20, a splicing factor, and CLIC1, an intracellular chloride ion channel, as novel downstream effectors besides previously reported effectors of Rho-guanine nucleotide dissociation inhibitor 2 and glutathione S-transferase-pi. Reduction in SRp20 was confirmed by one-dimensional Western blotting. Moreover, MG-132, a proteasome inhibitor, prevented this reduction. By contrast, upregulation of CLIC1 was not observed in one-dimensional Western blotting, unlike the 2-DE results. As hydrophilic proteins were predominantly recovered in 2-DE, the discrepancy between the 1-DE and 2-DE results may indicate a certain qualitative change of the protein. Indeed, the nuclear localization pattern of CLIC1 was remarkably changed by insulin stimulation. Thus insulin induces the proteasome dependent degradation of SRp20 as well as the subnuclear relocalization of CLIC1. PMID- 15827066 TI - Alterations in growth and apoptosis of insulin receptor substrate-1-deficient beta-cells. AB - Insulin and IGF-I activate antiapoptotic pathways via insulin receptor substrate (IRS) proteins in most mammalian cells, including beta-cells. IRS-1 knockout (IRS 1KO) mice show growth retardation, hyperinsulinemia, and hyperplastic but dysfunctional islets without developing overt diabetes, whereas IRS-2KOs develop insulin resistance and islet hypoplasia leading to diabetes. Because both models display insulin resistance, it is difficult to differentiate islet response to insulin resistance from islet defects due to loss of proteins in the islets themselves. We used a transplantation approach, as a means of separating host insulin resistance from islet function, to examine alterations in proteins in insulin/IGF-I signaling pathways that may contribute to beta-cell proliferation and/or apoptosis in IRS-1KO islets. Islets isolated from wild-type (WT) or IRS 1KO mice were transplanted into WT or insulin-resistant IRS-1KO males under the kidney capsule. The beta-cell mitotic rate in transplanted islets in IRS-1KO recipients was increased 1.5-fold compared with WT recipients and was similar to that in endogenous pancreases of IRS-1KOs, whereas beta-cell apoptosis was reduced by approximately 80% in IRS-1KO grafts in IRS-1KO recipients compared with WT recipients. Immunohistochemistry showed a substantial increase in IRS-2 expression in IRS-1KO islets transplanted into IRS-1KO mice as well as in endogenous islets from IRS-1KOs. Furthermore, enhanced cytosolic forkhead transcription factor (FoxO1) staining in IRS-1KO grafts suggests intact Akt/PKB activity. Together, these data indicate that, even in the absence of insulin resistance, beta-cells deficient in IRS-1 exhibit a compensatory increase in IRS 2, which is associated with islet growth and is characterized by both proliferative and antiapoptotic effects that likely occur via an insulin/IGF I/IRS-2 pathway. PMID- 15827067 TI - Trends in tuberculosis--US, 2004. PMID- 15827068 TI - Generalized seizure and toxic epidermal necrolysis following levofloxacin exposure. AB - OBJECTIVE: To report the case of a ciprofloxacin-allergic patient who developed a generalized tonic-clonic seizure and toxic epidermal necrolysis (TEN) following a single dose of levofloxacin. CASE SUMMARY: An 87-year-old white woman was admitted to the hospital following a transient episode of unresponsiveness that had been accompanied by flailing of her limbs. Approximately 4 hours earlier, she had developed a pruritic rash on her trunk and limbs, and 3 hours before this had taken a first dose of levofloxacin. The fluoroquinolone had been prescribed for treatment of an upper respiratory tract infection. She had developed a skin rash approximately 3 years earlier following ciprofloxacin prescribed for a urinary tract infection. On admission, the patient had a normal neurologic examination. She was mildly hypomagnesemic (serum magnesium 1.7 mg/dL), with no other electrolyte imbalances present. Skin biopsy confirmed TEN. The lesions progressed to involve 30% of the body surface area and were managed with polymyxin B and gramicidin cream. Levofloxacin was discontinued on admission, and no anticonvulsants were prescribed. The woman remained seizure-free at discharge one week later. DISCUSSION: Generalized tonic-clonic seizures are a rare complication of levofloxacin therapy. TEN following levofloxacin use has, to our knowledge, as of March 28, 2005, been previously reported only once. The seizure and TEN were probably induced by levofloxacin as corroborated by the Naranjo probability scale. We believe that the previous adverse dermatologic reaction to ciprofloxacin sensitized our patient to levofloxacin. CONCLUSIONS: These rare adverse reactions to levofloxacin, involving disparate organ systems, can occur simultaneously. A previous dematologic adverse reaction to a fluoroquinolone can sensitize a patient to more severe adverse reactions (with onset after only a single dose of the subsequent fluoroquinolone). Further fluoroquinolone use should be avoided in such patients. PMID- 15827070 TI - Sudden cardiac death due to risperidone therapy in a patient with possible hypertrophic cardiomyopathy. PMID- 15827069 TI - Etoricoxib: a highly selective COX-2 inhibitor. AB - OBJECTIVE: To review the available literature evaluating the pharmacology, pharmacokinetics, clinical efficacy, and adverse effects of etoricoxib, a highly selective cyclooxygenase-2 (COX-2) inhibitor that is not currently approved for use in the US. DATA SOURCES: Literature retrieval was accessed through MEDLINE (1966-December 2004), Current Contents (1998-December 2004), and Cochrane Library (4th quarter 2004). References from retrieved articles, information from the manufacturer, and abstracts from the American College of Rheumatology and Annual European Congress of Rheumatology meetings were searched. STUDY SELECTION AND DATA EXTRACTION: All clinical trials published in English evaluating etoricoxib were included in this review. An abstract was excluded if it presented preliminary data from trials that are now published, analyzed data previously reported in a published clinical trial, or compared etoricoxib with placebo for an indication with published active-comparator controlled trials. DATA SYNTHESIS: Twelve clinical trials evaluating efficacy were reviewed. Efficacy for acute pain has been evaluated in acute gout, primary dysmenorrhea, and dental surgery and for chronic pain in rheumatoid arthritis, osteoarthritis, and chronic lower back pain. For safety, 3 clinical trials and 6 retrospective analyses of gastrointestinal, renovascular, or cardiovascular adverse effects were reviewed. CONCLUSIONS: Available studies demonstrate the efficacy of etoricoxib compared with nonsteroidal antiinflammatory drugs, but no published studies to date have compared etoricoxib with other selective COX-2 inhibitors. While these agents have demonstrated a significant reduction in gastrointestinal adverse effects, the cardiovascular adverse effects of selective COX-2 inhibition are not well defined. Further study is necessary to delineate the benefits and risks of etoricoxib compared with alternative treatment regimens. PMID- 15827071 TI - Serotonin toxicity associated with concomitant use of linezolid. AB - OBJECTIVE: To report 2 cases of serotonin toxicity (ST) associated with concomitant use of linezolid and serotonergic drugs and review previously published case reports. CASE SUMMARIES: Case 1. A 38-year-old white female with cystic fibrosis treated with venlafaxine 300 mg/day for one year was prescribed linezolid 600 mg intravenously every 12 hours for treatment of methicillin resistant Staphylococcus aureus (MRSA) pulmonary infection. She displayed symptoms of ST 8 days after the introduction of linezolid. The venlafaxine dosage was decreased to 150 mg/day, and symptoms gradually abated over 36 hours. Case 2. A 37-year-old male with multiple myeloma received citalopram 40 mg/day and trazodone 150 mg/day for anxiety-related disorders. Linezolid treatment with 600 mg orally twice daily was instituted for MRSA cellulitis. The following day, the patient developed anxiety, panic attacks, tremors, tachycardia, and hypertension that persisted throughout linezolid treatment. Symptoms finally waned 5 days after linezolid treatment was stopped. DISCUSSION: The symptoms observed in our patients were consistent with Sternbach's criteria for ST. A review of published case reports showed a short time to onset of symptoms following the introduction of linezolid, generally within 1-3 days. Also of note is the use of relatively high dosages of serotonergic drugs. Use of the Naranjo probability scale indicated a possible relationship between the use of linezolid and the occurrence of ST in both cases. CONCLUSIONS: Clinicians should pay special attention to patients treated with serotonergic drugs, especially those receiving dosages in the higher end of the normal range who are prescribed linezolid, and consider tapering or reducing the dosage of serotonergic drugs for the duration of antibiotic therapy. PMID- 15827072 TI - Inhaled insulin: Exubera. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, efficacy, and safety of Exubera, a novel, dry-powder formulation of insulin for inhalation, and describe patient satisfaction and quality-of-life data. DATA SOURCES: A MEDLINE search (1966-November 2004) was conducted using the key words inhaled insulin and Exubera for clinical trials limited to human research published in English. BIOSIS Previews and the American Diabetes Association Scientific Abstracts were used for published abstract information. STUDY SELECTION AND DATA EXTRACTION: All available human studies of Exubera were selected for review. References of identified articles were used for additional citations. DATA SYNTHESIS: Exubera is a rapid-acting insulin administered by oral inhalation before meals with long acting insulin administered subcutaneously once or twice daily for type 1 or 2 diabetes mellitus. Exubera provides similar efficacy and improved patient satisfaction compared with standard subcutaneous insulin therapy (ie, NPH twice daily with regular insulin before meals). Efficacy has also been demonstrated for Exubera when used as adjunctive therapy with oral medications for type 2 diabetes. The onset of Exubera is more rapid and its duration of action is similar to that of regular insulin. To date, Exubera administered before meals with a once-daily long-acting subcutaneous insulin (usually Ultralente) has been compared with standard subcutaneous NPH/regular insulin regimens. Comparison of premeal Exubera plus a basal long-acting insulin analog (eg, glargine) with a regimen of premeal subcutaneous rapid-acting insulin analog (eg, lispro or aspart) plus a basal long-acting insulin analog (eg, glargine) is needed to fully evaluate Exubera. Pulmonary safety appears to be maintained for up to 4 years, although there are no data, as of this writing, on the use of this agent in patients with pulmonary conditions. CONCLUSIONS: Exubera is an effective inhaled insulin for preprandial use in type 1 or 2 diabetes. Improved patient satisfaction over injected insulin increases its potential for use earlier in the treatment of type 2 diabetes. PMID- 15827073 TI - Cefprozil-induced rash in infectious mononucleosis. PMID- 15827074 TI - Sildenafil for pulmonary hypertension. AB - OBJECTIVE: To evaluate the efficacy of sildenafil for treatment of pulmonary hypertension. DATA SOURCES: Literature retrieval was accessed through MEDLINE (1977-March 2005), Cochrane Library, and International Pharmaceutical Abstracts (1977-March 2005) using the terms sildenafil and pulmonary hypertension. In addition, reference citations from publications identified were reviewed. STUDY SELECTION AND DATA EXTRACTION: All articles in English identified from the data sources were evaluated. Studies including >5 patients with primarily adult populations were included in the review. DATA SYNTHESIS: The treatment of pulmonary hypertension is challenging. Sildenafil has recently been studied as monotherapy and in combination with other vasodilators in the management of pulmonary hypertension. Eight hemodynamic studies and 12 clinical trials were reviewed (1 retrospective, 3 double-blind, 8 open-label). Sildenafil reduced pulmonary arterial hypertension and pulmonary vascular resistance/peripheral vascular resistance index and tended to increase cardiac output/cardiac index compared with baseline. Sildenafil was comparable to nitric oxide and at least as effective as iloprost or epoprostenol in terms of its pulmonary vasoreactivity. Combination therapy with iloprost, nitric oxide, or epoprostenol resulted in enhanced and prolonged pulmonary vascular effects. Clinical trials suggest that sildenafil improves exercise tolerance and New York Heart Association functional class, but large, randomized controlled trials are needed to confirm these findings. Overall, sildenafil was well tolerated. CONCLUSIONS: Overall, sildenafil is a promising and well-tolerated agent for management of pulmonary hypertension. Further well-designed trials are warranted to establish its place in the treatment of pulmonary hypertension. PMID- 15827075 TI - Impact of a pharmaceutical care program in a community pharmacy on patients with dyslipidemia. AB - BACKGROUND: Inappropriate use of medications is a significant problem in health care today. A possible solution to this problem may be achieved through better control of patients' drug therapy. OBJECTIVE: To design a pharmaceutical care program for dyslipidemic patients within a community pharmacy setting that provides education in the areas of medication compliance and lifestyle modifications, while emphasizing the importance of achieving cholesterol goals to ensure improvement in quality of life. METHODS: Patients at an outpatient pharmacy volunteered to be surveyed for 16 weeks. Although both the intervention and control groups were surveyed, the randomly selected intervention group was interviewed more frequently and more comprehensively. Cholesterol, triglycerides, glucose, weight, risk factors, drug-related problems (DRPs), and quality of life were measured via a survey at the onset of the study and continually measured until the study's conclusion. RESULTS: In the intervention group, 26 DRPs were detected, of which 24 were resolved; in the control group, 26 DRPs were detected, of which 5 were resolved. When comparing initial and final blood cholesterol levels in the intervention group, the mean decrease was 27.0 +/- 41.1 mg/dL (p = 0.0266); in the control group, the average blood cholesterol level decreased by a mean of 1.4 +/- 37.2 mg/dL (p = 0.6624). In the intervention group, the triglyceride level decreased an average of 50.5 +/- 80.3 mg/dL (p = 0.0169), while the control group experienced a mean triglyceride level increase of 29.6 +/ 118.5 mg/dL (p = 0.1435). As a result of the intervention, the quality of life in the intervention group was improved. CONCLUSIONS: Short-term pharmaceutical care plans developed in a retail pharmacy within the proper setting may contribute to improved blood lipid values, cardiovascular disease risk factors, and patients' quality of life. PMID- 15827076 TI - Comment: urinary tract infections during pregnancy. PMID- 15827077 TI - A prospective study of psychosocial risk factors and absence due to musculoskeletal disorders--implications for occupational screening. AB - OBJECTIVES: There is considerable evidence documenting the association between psychosocial risk factors and work disability due to musculoskeletal disorders, and this has prompted suggestions that psychosocial screening should be administered in the workplace in order to identify individuals at risk of prolonged absence. However, the predictive value of psychosocial risk factors on return-to-work is largely unknown. The present study aimed to explore the predictive relationship between psychosocial risk factors and absence due to musculoskeletal disorders of the lower back and upper limbs. METHODS: A prospective study of 4637 workers from a large, multi-site company in the UK was conducted in which a wide range of established questionnaires were used to collect baseline psychosocial data. Respondents were then followed over the ensuing 15 months, and absence due to musculoskeletal disorders was recorded. RESULTS: 219 workers took absence due to musculoskeletal disorders. Detrimental cut-off scores (risks) on the psychosocial instruments were established, and it was found that work-related psychosocial risk factors predicted the likelihood of a future spell of absence (odds ratios ranging between 1.6 and 3.2), but not the duration of that absence. CONCLUSIONS: Although work-related psychosocial factors were associated with the occurrence of absence due to musculoskeletal disorders, these findings do not lend support to the use of routine occupational psychosocial screening in order to predict prolonged absence. PMID- 15827078 TI - STARS: statistics on inter-atomic distances and torsion angles in protein secondary structures. AB - A graphics package has been developed for performing statistics on interatomic distances and torsion angles in protein secondary structures (STARS) from a protein crystal structure database. It allows one to obtain both the graphical view and the text format of distributions of the distances and angles for atoms located in 10 types of protein secondary structures. STARS will facilitate assignment of ambiguous NOESY peaks, structure determination by nuclear magnetic resonance, structure validation and comparison of protein folds. AVAILABILITY: All data, documents and execute files are freely downloadable at http://stars.zhengyuhome.com. The software works appropriately on Windows system, without any compilation or installation. CONTACT: dbsydw@nus.edu.sg. PMID- 15827079 TI - Visualizing profile-profile alignment: pairwise HMM logos. AB - The availability of advanced profile-profile comparison tools, such as PRC or HHsearch demands sophisticated visualization tools not presently available. We introduce an approach built upon the concept of HMM logos. The method illustrates the similarities of pairs of protein family profiles in an intuitive way. Two HMM logos, one for each profile, are drawn one upon the other. The aligned states are then highlighted and connected. AVAILABILITY: A web interface offering online creation of pairwise HMM logos is available at http://www.sanger.ac.uk/Software/analysis/logomat-p. Furthermore, software developers may download a Perl package that includes methods for creation of pairwise HMM logos locally. CONTACT: bsb@sanger.ac.uk. PMID- 15827080 TI - Three-dimensional computation of atom depth in complex molecular structures. AB - MOTIVATION: For a complex molecular system the delineation of atom-atom contacts, exposed surface and binding sites represents a fundamental step to predict its interaction with solvent, ligands and other molecules. Recently, atom depth has been also considered as an additional structural descriptor to correlate protein structure with folding and functional properties. The distance between an atom and the nearest water molecule or the closest surface dot has been proposed as a measure of the atom depth, but, in both cases, the 3D character of depth is largely lost. In the present study, a new approach is proposed to calculate atom depths in a way that the molecular shape can be taken into account. RESULTS: An algorithm has been developed to calculate intersections between the molecular volume and spheres centered on the atoms whose depth has to be quantified. Many proteins with different size and shape have been chosen to compare the results obtained from distance-based and volume-based depth calculations. From the wealth of experimental data available for hen egg white lysozyme, H/D exchange rates and TEMPOL induced paramagnetic perturbations have been analyzed both in terms of depth indexes and of atom distances to the solvent accessible surface. The algorithm here proposed yields better correlations between experimental data and atom depth, particularly for those atoms which are located near to the protein surface. AVAILABILITY: Instructions to obtain source code and the executable program are available either from http://sienabiografix.com or http://sadic.sourceforge.net CONTACT: niccolai@unisi.it SUPPLEMENTARY INFORMATION: http://www.Sienabiogzefix.com/publication. PMID- 15827082 TI - EMPReSS: European mouse phenotyping resource for standardized screens. AB - Standardized phenotyping protocols are essential for the characterization of phenotypes so that results are comparable between different laboratories and phenotypic data can be related to ontological descriptions in an automated manner. We describe a web-based resource for the visualization, searching and downloading of standard operating procedures and other documents, the European Mouse Phenotyping Resource for Standardized Screens-EMPReSS. AVAILABILITY: Direct access: http://www.empress.har.mrc.ac.uk CONTACT: e.green@har.mrc.ac.uk. PMID- 15827081 TI - LS-SNP: large-scale annotation of coding non-synonymous SNPs based on multiple information sources. AB - MOTIVATION: The NCBI dbSNP database lists over 9 million single nucleotide polymorphisms (SNPs) in the human genome, but currently contains limited annotation information. SNPs that result in amino acid residue changes (nsSNPs) are of critical importance in variation between individuals, including disease and drug sensitivity. RESULTS: We have developed LS-SNP, a genomic scale software pipeline to annotate nsSNPs. LS-SNP comprehensively maps nsSNPs onto protein sequences, functional pathways and comparative protein structure models, and predicts positions where nsSNPs destabilize proteins, interfere with the formation of domain-domain interfaces, have an effect on protein-ligand binding or severely impact human health. It currently annotates 28,043 validated SNPs that produce amino acid residue substitutions in human proteins from the SwissProt/TrEMBL database. Annotations can be viewed via a web interface either in the context of a genomic region or by selecting sets of SNPs, genes, proteins or pathways. These results are useful for identifying candidate functional SNPs within a gene, haplotype or pathway and in probing molecular mechanisms responsible for functional impacts of nsSNPs. AVAILABILITY: http://www.salilab.org/LS-SNP CONTACT: rachelk@salilab.org SUPPLEMENTARY INFORMATION: http://salilab.org/LS-SNP/supp-info.pdf. PMID- 15827083 TI - Selective Golgi export of Kir2.1 controls the stoichiometry of functional Kir2.x channel heteromers. AB - Surface expression of ion channels and receptors often depends on intrinsic sequence motifs that control their intracellular transport along the secretory pathway. Although members of the Kir2.x subfamily share two such motifs - a diacidic ER export motif and a positively charged Golgi export motif - they strongly differ in their surface expression. Whereas Kir2.1 shows prominent plasma membrane localization, Kir2.4 channels accumulate within the Golgi complex. By constructing chimeras between Kir2.1 and Kir2.4 subunits, a stretch of 20 amino acids was identified in the Kir2.1 C-terminus that is both necessary and sufficient to promote anterograde transport of Kir channel subunits at the level of trafficking from the Golgi to the plasma membrane. The core element of the identified sequence bears a tyrosine-dependent YXXPhi consensus motif for adaptin binding, with the flanking residues determining its functional efficiency. As the signal is dominant in promoting surface transport of Kir2.1/Kir2.4 channel heteromers and is recognized by both the epithelial and neuronal intracellular sorting machinery, the preferential Golgi export of Kir2.1 will control the stoichiometry of Kir2.x heteromers expressed on the cell surface. PMID- 15827084 TI - RasGEF-containing proteins GbpC and GbpD have differential effects on cell polarity and chemotaxis in Dictyostelium. AB - The regulation of cell polarity plays an important role in chemotaxis. Previously, two proteins termed GbpC and GbpD were identified in Dictyostelium, which contain RasGEF and cyclic nucleotide binding domains. Here we show that gbpC-null cells display strongly reduced chemotaxis, because they are unable to polarise effectively in a chemotactic gradient. However, gbpD-null mutants exhibit the opposite phenotype: cells display improved chemotaxis and appear hyperpolar, because cells make very few lateral pseudopodia, whereas the leading edge is continuously remodelled. Overexpression of GbpD protein results in severely reduced chemotaxis. Cells extend many bifurcated and lateral pseudopodia, resulting in the absence of a leading edge. Furthermore, cells are flat and adhesive owing to an increased number of substrate-attached pseudopodia. This GbpD phenotype is not dependent on intracellular cGMP or cAMP, like its mammalian homolog PDZ-GEF. Previously we showed that GbpC is a high-affinity cGMP binding protein that acts via myosin II. We conclude that cGMP activates GbpC, mediating the chemoattractant-induced establishment of cell polarity through myosin. GbpD induces the formation of substrate-attached pseudopodia, resulting in increased attachment and suppression of polarity. PMID- 15827085 TI - PAK4 mediates morphological changes through the regulation of GEF-H1. AB - Precise spatial and temporal regulation of Rho GTPases is required in controlling F-actin-based changes in cell morphology. The molecular mechanisms through which microtubules (MTs) modulate the activity of RhoGTPases and regulate the actin cytoskeleton are unclear. Here we show that p21-activated-kinase 4 (PAK4) mediates morphological changes through its association with the Rho-family guanine nucleotide exchange factor (GEF), GEF-H1. We show that this association is dependent upon a novel GEF-H1 interaction domain (GID) within PAK4. Further, we show that PAK4-mediated phosphorylation of Ser810 acts as a switch to block GEF-H1-dependent stress fiber formation while promoting the formation of lamellipodia in NIH-3T3 cells. We found that the endogenous PAK4-GEF-H1 complex associates with MTs and that PAK4 phosphorylation of MT-bound GEF-H1 releases it into the cytoplasm of NIH-3T3 cells, which coincides with the dissolution of stress fibers. Our observations propose a novel role for PAK4 in GEF-H1-dependent crosstalk between MTs and the actin cytoskeleton. PMID- 15827086 TI - Nef induces apoptosis by activating JNK signaling pathway and inhibits NF-kappaB dependent immune responses in Drosophila. AB - The human immunodeficiency virus type 1 (HIV-1) nef gene encodes a 27-kDa protein that plays a crucial role during AIDS pathogenesis, but its exact functional mechanism has not been fully elucidated and remains controversial. The present study illuminated the in vivo functions of Nef using Drosophila, in which genetic analyses can be conveniently conducted. Using Drosophila transgenic lines for wild-type Nef, we demonstrated that Nef is not involved in the regulation of cell proliferation but rather specifically induces caspase-dependent apoptosis in wings in a cell-autonomous manner. Interestingly, myristoylation-defective Nef completely failed to induce the apoptotic wing phenotypes, consistent with previous reports demonstrating a crucial role for membrane localization of Nef in vivo. Further genetic and immunohistochemical studies revealed that Nef-dependent JNK activation is responsible for apoptosis. Furthermore, we found that ectopic expression of Nef inhibits Drosophila innate immune responses including Relish NF kappaB activation with subsequent induction of an antimicrobial peptide, diptericin. The in vivo functions of Nef in Drosophila are highly consistent with those found in mammals and so we propose that Nef regulates evolutionarily highly conserved signaling molecules of the JNK and NF-kappaB signaling pathways at the plasma membrane, and consequently modulates apoptosis and immune responses in HIV target cells. PMID- 15827087 TI - End4/Sla2 is involved in establishment of a new growth zone in Schizosaccharomyces pombe. AB - The rod-shaped Schizosaccharomyces pombe cell grows in a polarized fashion from opposing ends. Correct positioning of the growth zones is directed by the polarity marker Tea1 located at the cell ends where actin patches accumulate and cell growth takes place. We show that the S. pombe homologue of Saccharomyces cerevisiae SLA2, a protein involved in cortical actin organization and endocytosis, provides a link between the polarity marker and the growth machinery. In wild-type fission yeast cells, this homologue End4/Sla2 is enriched at cell ends during interphase and localizes to a medial ring at cell division, mirroring the actin localization pattern throughout the cell cycle. Proper localization relies on membrane trafficking and is independent of both the actin and microtubule cytoskeletons. End4/Sla2 is required for the establishment of new polarised growth zones, and deletion of its C-terminal talin-like domain prevents the establishment of a new growth zone after cell fission. We propose that End4/Sla2 acts downstream of the polarity marker Tea1 and is implicated in the recruitment of the actin cytoskeleton to bring about polarised cell growth. PMID- 15827088 TI - p130/p107/p105Rb-dependent transcriptional repression during DNA-damage-induced cell-cycle exit at G2. AB - The progression of normal cells from G2 into mitosis is stably blocked when their DNA is damaged. Tumor cells lacking p53 arrest only transiently in G2, but eventually enter mitosis. We show that an important component of the stable G2 arrest in normal cells is the transcriptional repression of more than 20 genes encoding proteins needed to enter into and progress through mitosis. Studies from a number of labs including our own have shown that, by inducing p53 and p21/WAF1, DNA damage can trigger RB-family-dependent transcriptional repression. Our studies reported here show that p130 and p107 play a key role in transcriptional repression of genes required for G2 and M in response to DNA damage. For plk1, repression is partially abrogated by loss of p130 and p107, and is completely abrogated by loss of all three RB-family proteins. Mouse cells lacking RB-family proteins do not accumulate with a 4N content of DNA when exposed to adriamycin, suggesting that all three RB-family proteins contribute to G2 arrest in response to DNA damage. Stable arrest in the presence of functional p53-to-RB signaling is probably due to the ability of cells to exit the cell cycle from G2, a conclusion supported by our observation that KI67, a marker of cell-cycle entry, is downregulated in both G1 and G2 in a p53-dependent manner. PMID- 15827089 TI - Non-random chromosome positioning in mammalian sperm nuclei, with migration of the sex chromosomes during late spermatogenesis. AB - Chromosomes are highly organized and compartmentalized in cell nuclei. The analysis of their position is a powerful way to monitor genome organization in different cell types and states. Evidence suggests that the organization of the genome could be functionally important for influencing different cellular and developmental processes, particularly at early stages of development (i.e. fertilization and the consequent entry of the sperm nucleus into the egg). The position of chromosomes in the sperm nucleus might be crucial, because their location could determine the time at which particular chromatin domains are decondensed and remodelled, allowing some epigenetic level of control or influence over subsequent paternal gene expression in the embryo. Here, we analyse genome organization by chromosome position in mammalian sperm nuclei from three breeds of pig, as a model species. We have mapped the preferential position of all chromosomes (bar one) in sperm nuclei in two dimensions and have established that the sex chromosomes are the most internally localized chromosomes in mature sperm. The distribution of two autosomes and chromosomes X and Y in sperm heads was compared in primary and secondary spermatocytes and spermatids in porcine testes. The sex chromosomes were found at the nuclear edge in primary spermatocytes, which correlates with the known position of the XY body and their position in somatic cells, whereas, in spermatids, the sex chromosomes were much more centrally located, mirroring the position of these chromosomes in ejaculated spermatozoa. This study reveals the temporal repositioning of chromosome territories in spermatogenesis. PMID- 15827090 TI - Mechanics of neutrophil phagocytosis: behavior of the cortical tension. AB - The mechanical implementation of phagocytosis requires a well-coordinated deployment of cytoplasm and membrane during the creation of a phagosome. We follow the time course of this process in initially round passive neutrophils presented with antibody-coated beads of radii 1.1 to 5.5 microm. In particular, we monitor the cortical tension as the apparent cellular surface area increases due to cell-driven deformations induced by phagocytosis. The behavior of the tension is then compared with conditions of similar area expansion caused by externally imposed deformations during cell aspiration into a micropipette. Whereas the resting tension remains low for an area expansion of up to only 30% during aspiration, it remains low even after an area expansion of up to 80% in phagocytosis. This is probably the result of membrane insertion from inner stores by exocytosis. We further find that the onset of viscous tension, proportional to the rate of area expansion and caused by the unfurling of plasma membrane wrinkles, is significantly delayed in phagocytosis compared with aspiration. We propose that this is the result of phagocytosis-triggered enzymatic activity that releases spare plasma membrane normally sequestered by velcro-like bonds in a reservoir of surface folds and villi. PMID- 15827091 TI - A novel susceptibility locus at 2p24 for generalised epilepsy with febrile seizures plus. AB - Generalised epilepsy with febrile seizures plus (GEFS+) is a clinically and genetically heterogeneous epilepsy syndrome. Using positional cloning strategies, mutations in SCN1B, SCN1A, and GABRG2 have been identified as genetic causes of GEFS+. In the present study, we describe a large four generation family with GEFS+ in which we performed a 10 cM density genome-wide scan. We obtained conclusive evidence for a novel GEFS+ locus on chromosome 2p24 with a maximum two point logarithm of the odds (LOD) score of 4.22 for marker D2S305 at zero recombination. Fine mapping and haplotype segregation analysis in this family delineated a candidate region of 3.24 cM, corresponding to a physical distance of 4.2 Mb. Linkage to 2p24 was confirmed (p = 0.007) in a collection of 50 nuclear and multiplex families with febrile seizures and epilepsy. Transmission disequilibrium testing and association studies provided further evidence (p < 0.05) that 2p24 is a susceptibility locus for febrile seizures and epilepsy. Furthermore, we could reduce the candidate region to a 2.14 cM interval, localised between D2S1360 and D2S2342, based upon an ancestral haplotype. Identification of the disease gene at this locus will contribute to a better understanding of the complex genetic aetiology of febrile seizures and epilepsy. PMID- 15827092 TI - A male-specific quantitative trait locus on 1p21 controlling human stature. AB - BACKGROUND: Many genome-wide scans aimed at complex traits have been statistically underpowered due to small sample size. Combining data from several genome-wide screens with comparable quantitative phenotype data should improve statistical power for the localisation of genomic regions contributing to these traits. OBJECTIVE: To perform a genome-wide screen for loci affecting adult stature by combined analysis of four previously performed genome-wide scans. METHODS: We developed a web based computer tool, Cartographer, for combining genetic marker maps which positions genetic markers accurately using the July 2003 release of the human genome sequence and the deCODE genetic map. Using Cartographer, we combined the primary genotype data from four genome-wide scans and performed variance components (VC) linkage analyses for human stature on the pooled dataset of 1417 individuals from 277 families and performed VC analyses for males and females separately. RESULTS: We found significant linkage to stature on 1p21 (multipoint LOD score 4.25) and suggestive linkages on 9p24 and 18q21 (multipoint LOD scores 2.57 and 2.39, respectively) in males-only analyses. We also found suggestive linkage to 4q35 and 22q13 (multipoint LOD scores 2.18 and 2.85, respectively) when we analysed both females and males and to 13q12 (multipoint LOD score 2.66) in females-only analyses. CONCLUSIONS: We strengthened the evidence for linkage to previously reported quantitative trait loci (QTL) for stature and also found significant evidence of a novel male specific QTL on 1p21. Further investigation of several interesting candidate genes in this region will help towards characterisation of this first sex specific locus affecting human stature. PMID- 15827093 TI - Severe growth hormone insensitivity resulting from total absence of signal transducer and activator of transcription 5b. AB - CONTEXT: The central clinical feature of GH insensitivity (GHI) is severe growth failure associated with elevated serum concentrations of GH and abnormally low serum levels of IGF-I. GHI can be the result of an abnormality in the GH receptor or aberrancies downstream of the GH receptor. OBJECTIVE: We investigated the GH IGF-I axis in a young female GHI subject who presented with a height of 114 cm ( 7.8 sd score) at age 16.4 yr. PATIENT: The subject, from a consanguineous pedigree, had circulating levels of GH and GH-binding protein that were normal to elevated, whereas IGF-I (7.2 ng/ml; normal, 242-600), IGF-binding protein-3 (543 ng/ml; normal, 2500-4800), and acid-labile subunit (1.22 microg/ml; normal, 5.6 16) levels were abnormally low and failed to increase during an IGF-I generation test. DESIGN: Dermal fibroblast cultures were established with the consent of the patient and family. Immunoblot analysis of cell lysates and DNA sequencing of her signal transducer and activator of transcription 5b (STAT5b), a critical intermediate of the GH-IGF-I axis, were performed. RESULTS: Sequencing of the STAT5b gene revealed a novel homozygous insertion of a single nucleotide in exon 10. The insertion resulted in a frame shift, leading to early protein termination and consequent lack of immunodetectable STAT5b protein. CONCLUSION: The identification of a second case of severe growth failure associated with STAT5b mutation implicates a unique and critical role for STAT5b in GH stimulation of IGF-I gene expression and statural growth. PMID- 15827094 TI - Dose-dependent effects of testosterone on sexual function, mood, and visuospatial cognition in older men. AB - CONTEXT: The relationships between testosterone dose and its effects on sexual function, mood, and visuospatial cognition are poorly understood. OBJECTIVE: To elucidate testosterone dose-response relationships in older men, we examined the effects of graded testosterone doses on sexual function, mood, and visuospatial cognition in healthy, older men (age, 60-75 yr). SETTING: This study was performed at the General Clinical Research Center. INTERVENTION/METHODS: Subjects each received a long-acting GnRH agonist to suppress endogenous testosterone production and were randomized to receive one of five doses (25, 50, 125, 300, and 600 mg) of testosterone enanthate weekly for 20 wk. Questionnaires were used to evaluate sexual function. Scores for overall sexual function as well as subcomponents of sexual function (libido, sexual activity, and erectile function) were calculated. RESULTS: Changes in overall sexual function (P = 0.003) and waking erections (P = 0.024) differed by dose. An interaction between libido and being sexually active was observed, such that libido changed by testosterone dose only among men who reported being sexually active at the beginning of the study (P = 0.009). Men's log-transformed free testosterone levels during treatment were positively correlated with overall sexual function (P = 0.001), waking erections (P = 0.040), spontaneous erections (P = 0.047), and libido (P = 0.027), but not with intercourse frequency (P = 0.428) or masturbation frequency (P = 0.814). No effects of testosterone dose were observed on two measures of mood: Hamilton's Depression Inventory (P = 0.359) and Young's Mania Scale (P = 0.851). The number of trials completed on a computer-based test of visuospatial cognition differed by dose (P = 0.042), but the number of squares correctly completed on this task did not differ by dose (P = 0.159). CONCLUSIONS: Different aspects of male behavior respond differently to testosterone. When considered together with previous data from young men, these data indicate that testosterone dose-response relationships for sexual function and visuospatial cognition differ in older and young men. PMID- 15827095 TI - Androgen levels in adult females: changes with age, menopause, and oophorectomy. AB - CONTEXT: Changes in androgen levels across the adult female life span and the effects of natural menopause and oophorectomy have not been clearly established. OBJECTIVE: The objective of this study was to document the effects of age on androgen levels in healthy women and to explore the effects of natural and surgical menopause. DESIGN, SETTING, AND PARTICIPANTS: A cross-sectional study was conducted of 1423 non-healthcare-seeking women, aged 18-75 yr, randomly recruited from the community over 15 months. MAIN OUTCOME MEASURES: Serum levels by age of total testosterone (T), calculated free T, dehydroepiandrosterone sulfate, and androstenedione in a reference group of women free of confounding factors. Women in the reference group had no usage of exogenous steroid therapy; no history of tubal ligation, hysterectomy, or bilateral oophorectomy; and no hyperprolactinemia or polycystic ovarian syndrome. The effects of natural and surgical menopause on sex steroid levels were also examined. RESULTS: In the reference population (n = 595), total T, calculated free T, dehydroepiandrosterone sulfate, and androstenedione declined steeply with age (P < 0.001), with the decline of each being greater in the earlier than the later decades. Examination of serum androgen levels by year in women aged 45-54 yr showed no independent effect of menopausal status on androgen levels. In women aged 55 yr or older, those who reported bilateral oophorectomy and were not on exogenous steroids had significantly lower total T and free T levels than women 55 yr or older in the reference group. CONCLUSIONS: We report that serum androgen levels decline steeply in the early reproductive years and do not vary because a consequence of natural menopause and that the postmenopausal ovary appears to be an ongoing site of testosterone production. These significant variations in androgens with age must be taken into account when normal ranges are reported and in studies of the role of androgens in women. PMID- 15827096 TI - Disturbance of the fetal thyroid hormone state has long-term consequences for treatment of thyroidal and central congenital hypothyroidism. AB - BACKGROUND: During T(4) supplementation of patients with thyroidal (primary) congenital hypothyroidism (CH) TSH concentrations are frequently elevated despite free T(4) (FT(4)) concentrations being well within the reference range. To examine the thyroid's regulatory system, we analyzed thyroid function determinants in children with congenital and acquired thyroid disorders and in controls. METHODS: Retrospectively, plasma FT(4), TSH, and T(3) concentrations were analyzed in T(4)-supplemented children aged 0.5-20.0 yr with thyroidal CH, central (secondary or tertiary) CH, or autoimmune thyroid disease and in control children with type 1 diabetes mellitus. RESULTS: When TSH was within the reference range (0.4-4.0 mU/liter), mean FT(4) in thyroidal CH [1.65 ng/dl; 95% confidence interval (CI), 1.62-1.67] was significantly higher than in autoimmune thyroid disease (1.15 ng/dl; 95% CI, 1.11-1.19) and diabetes (1.08 ng/dl; 95% CI, 1.06-1.10). In central CH, when TSH was less than or equal to 0.02 mU/liter, mean FT(4) was 1.27 ng/dl (95% CI, 1.24-1.29). When FT(4) was within the reference range (0.78-1.79 ng/dl), 43% of the TSH measurements in thyroidal CH were more than 4.0 mU/liter, compared with 18% in autoimmune thyroid disease and 0% in type 1 diabetes mellitus; in central CH, 95% of TSH measurements were less than 0.4 mU/liter. CONCLUSIONS: In T(4)-supplemented patients with thyroidal CH, when TSH concentrations are established within the reference range, FT(4) concentrations tend to be elevated, and vice versa. Because this phenomenon could not be observed in acquired thyroidal hypothyroidism, we hypothesize that a pre- and/or perinatal hypothyroid state shifts the setpoint of the thyroid's regulatory system. In central CH, when FT(4) concentrations are established within the reference range, the pituitary secretes only minute amounts of TSH. For monitoring T(4) supplementation, reference ranges for FT(4) and TSH should be adapted to the etiology of hypothyroidism. PMID- 15827097 TI - Codon-specific development of pheochromocytoma in multiple endocrine neoplasia type 2. AB - CONTEXT: Recent data suggest a codon-specific, age-related development of multiple endocrine neoplasia type 2. OBJECTIVE: The objective of this study was to delineate the codon-specific, age-related development of multiple endocrine neoplasia type 2-associated pheochromocytoma. DESIGN: We describe a cohort study with a mean observation period of 26.9 yr. SETTING: The study took place in a tertiary referral center at a university hospital. PATIENTS: Included in this study were 206 consecutive carriers (74 index, 132 nonindex) operated on at this institution who harbored point mutations in the RET (rearranged during transfection) protooncogene. INTERVENTION: The intervention was adrenalectomy for clinically confirmed pheochromocytoma. MAIN OUTCOME MEASURE: The main outcome measure was time to histopathological diagnosis of pheochromocytoma. RESULTS: Pheochromocytomas developed in 28% (five of 18) of carriers with mutations in codon 918, 29% (20 of 68) of carriers with mutations in codon 634, 14% (three of 21) of carriers with mutations in codon 618, 13% (two of 16) of carriers with mutations in codon 620, and 13% (two of 16) of carriers with mutations in codon 791. Earliest age of manifestation for each genotype was 22, 18, 29, 22, and 39 yr. Contralateral pheochromocytomas developed after 4 yr (one carrier each had a mutation in codon 618 or 620) and 5.2 yr (six carriers had mutations in codon 634). No pheochromocytomas were identified in carriers of mutations in codons 609 (n = 2), 611 (n = 8), 630 (n = 2), 768 (n = 8), 790 (n = 22), 804 (n = 18), and 891 (n = 7). CONCLUSIONS: Based on these and published preliminary data, annual screening for pheochromocytoma may be warranted from age 10 yr in carriers of RET mutations in codons 918, 634, and 630, and from age 20 yr in the remainder. PMID- 15827098 TI - Adrenal ganglioneuromas in children with multiple endocrine neoplasia type 2: a report of two cases. AB - CONTEXT: Pheochromocytomas of the adrenal gland are a common component of the multiple endocrine neoplasia type 2 (MEN2) syndromes. However, pure adrenal ganglioneuromas, an extremely rare pediatric tumor of neural crest origin composed of mature ganglion cells, have never been reported in association with MEN2 in humans. MEN2A is comprised of medullary thyroid carcinoma (MTC), pheochromocytoma, and parathyroid hyperplasia. MEN2B is characterized by MTC, pheochromocytoma, neural abnormalities of the gastrointestinal tract, and mucosal neuromas. EVIDENCE ACQUISITION: We report two pediatric patients, one with MEN2A and one with MEN2B, who developed isolated adrenal ganglioneuromas without evidence of pheochromocytomas. EVIDENCE SYNTHESIS: MEN2A and MEN2B are caused by activating mutations in the RET proto-oncogene, which encodes a tyrosine kinase receptor essential for signal transduction in neural crest-derived tissues, including the peripheral and enteric nervous systems, C cells of the thyroid gland, and chromaffin cells of the adrenal gland. Both pheochromocytomas and ganglioneuromas originate from neural crest cells. Interestingly, two mouse models of MEN2B exhibit adrenal ganglioneuroma formation. One mouse model develops only ganglioneuromas (but not pheochromocytomas) and expresses only one of the oncogenic RET isoforms. The other mouse model, created by site-directed mutagenesis to simulate the most common human mutation, develops both ganglioneuromas and pheochromocytomas. CONCLUSIONS: Given our two cases, our current understanding of the mouse models, and the common origins of all these tumor cell types, we recommend including ganglioneuromas as a rare, but not unexpected, component of the MEN2 syndromes. PMID- 15827099 TI - Efficacy of octreotide-LAR in dieting women with abdominal obesity and polycystic ovary syndrome. AB - CONTEXT: Somatostatin reduces LH, GH, and insulin, and somatostatin receptors are present at the ovarian level; somatostatin analogs are thus potential candidates for treatment of the polycystic ovary syndrome (PCOS). OBJECTIVE: The purpose of this study was to evaluate the effect of octreotide-LAR, a long-acting somatostatin analog, in anovulatory abdominal obese women with PCOS. DESIGN: A single-blind, placebo-controlled study was performed, lasting for 7 months. SETTING: The patients were ambulatory throughout the study. PATIENTS: Twenty PCOS subjects were enrolled. Eighteen completed the study. INTERVENTIONS: A low calorie diet was given during the first month, a low-calorie diet plus octreotide LAR (10 mg; n = 10 subjects) or placebo (n = 10 subjects) was then given, with one im injection every 28 d (for 6 months). MAIN OUTCOME MEASURES: The main outcome measures were clinical features, computerized tomography measurement of fat distribution, androgens, GH, IGF-I, IGF-binding proteins (IGFBPs), fasting and glucose-stimulated insulin, and ovulation. RESULTS: Octreotide had no additional effect in reducing body fat or improving fat distribution than placebo. Conversely, octreotide produced an additional decrease in fasting (P = 0.018) and glucose-stimulated (P = 0.038) insulin levels, an increase in IGFBP-2 (P = 0.042) and IGFBP-3 (P = 0.047), and an improvement in hirsutism (P = 0.004). Moreover, a trend toward greater reductions in testosterone (P = 0.061) and androstenedione (P = 0.069) was observed in women treated with octreotide-LAR compared with those given placebo. All women treated with octreotide ovulated at the end of the study compared with only one of those receiving placebo (P < 0.001). CONCLUSIONS: Octreotide-LAR may be usefully applied to hypocalorically dieting, abdominal obese PCOS women to improve hyperandrogenism and the insulin IGF-I system. Restoration of ovulatory menstrual cycles appears to be another advantage of this treatment. PMID- 15827100 TI - Genetic and environmental factors influencing fasting serum adiponectin in Hispanic children. AB - CONTEXT: Because of its antiinflammatory and insulin-sensitizing properties, adiponectin may play a role in the development of cardiovascular disease and type 2 diabetes. OBJECTIVES: The aims of these analyses were: 1) to estimate the heritability of fasting serum adiponectin; 2) to evaluate the effects of age, sex, and body composition on fasting serum adiponectin; 3) to test for associations between fasting serum adiponectin and diet, fitness, energy expenditure, and fat oxidation; and 4) to determine the relationships between fasting serum adiponectin, insulin and lipids, and blood pressure in Hispanic children. DESIGN: Genetic and environmental factors influencing fasting serum adiponectin were investigated in a cohort of children participating in the VIVA LA FAMILIA Study in 2000-2005. SETTING: This study was performed at the Children's Nutrition Research Center. PARTICIPANTS: The study participants were 805 Hispanic nonoverweight and overweight children, ages 4-19 yr. MAIN MEASURE: The main measure of the study was fasting serum adiponectin. RESULTS: The heritability of serum adiponectin was 0.93 +/- 0.10 (P = 2.4 x 10(-40)). Adiponectin differed by age (P = 0.001), sex (P = 0.04), and weight (P = 0.001) status. Adiponectin levels declined with age, in association with changes in sex hormones and growth factors. Adiponectin was not associated with macronutrient intake, fitness, 24-h energy expenditure, or fat oxidation. Controlling for age, sex, and percent fat mass, adiponectin was inversely associated with homeostasis model of insulin resistance, triglycerides (TG)/high-density lipoprotein cholesterol (HDL-C), and systolic blood pressure (P = 0.001). Significant positive genetic correlations were detected between adiponectin and total cholesterol (rho(G) = 0.19), HDL-C (rho(G) = 0.32), low-density lipoprotein cholesterol (rho(G) = 0.24), and IGF-binding protein-1 (rho(G) = 0.39), and negative genetic correlations were detected between adiponectin and leptin (rho(G) = -0.30), TG (rho(G) = -0.21), TG/HDL-C (rho(G) = -0.33), and IGF-binding protein-3 (rho(G) = -0.32), indicating shared genetic components in their expression. CONCLUSION: The high heritability of adiponectin and pleiotropy seen between adiponectin and leptin, growth factors, and lipids may play a role in the pathogenesis of cardiovascular disease and type 2 diabetes in overweight Hispanic children. PMID- 15827101 TI - Peroxisome proliferator-activated receptor-gamma and retinoid X receptor signaling regulate fatty acid uptake by primary human placental trophoblasts. AB - CONTEXT: Transplacental transfer of fatty acids from the maternal to the fetal circulation is essential for fetal development. The nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) regulates fatty acid transport and storage in adipocytes and other cell types. OBJECTIVE: This study tested the hypothesis that PPARgamma and its heterodimeric nuclear receptor partner, retinoid X receptor (RXR), regulate fatty acid uptake by human trophoblasts. DESIGN: Prospective basic laboratory in vitro research was conducted using primary term human trophoblasts. SETTING: The study was performed in the perinatal biology laboratory of an academic medical center. PATIENTS OR OTHER PARTICIPANTS: Study materials were obtained from healthy pregnant women at a gestational age of 37-41 wk. INTERVENTIONS: There were no interventions. MAIN OUTCOME MEASURES: Fat uptake and accumulation in human placental trophoblasts were measured. RESULTS: We initially demonstrated that activation of PPARgamma and/or RXR with selective agonists increased the accumulation of neutral lipids in trophoblasts as well as uptake of free fatty acids. Furthermore, activation of PPARgamma and RXR enhanced the expression of the fat droplet-associated protein adipophilin along with fatty acid transport protein (FATP)4, whereas expression of FATP2 was decreased by activation of RXR. Finally, we found that inhibition of p38 MAPK, which diminishes the activity of PPARgamma in trophoblasts, inhibited fatty acid uptake and blocked the PPARgamma- and RXR-dependent increases in adipophilin and FATP4 expression, yet stimulated the expression of FATP1, FATP2, and FATP3. CONCLUSIONS: These data support a role for PPARgamma and RXR in regulation of fatty acid transport and storage in human placental trophoblasts. PMID- 15827102 TI - Ferriman Gallwey self-scoring I: performance assessment in women with polycystic ovary syndrome. AB - OBJECTIVE: The objective of this study was to determine performance characteristics of Ferriman Gallwey self-scoring in women with the polycystic ovary syndrome. DESIGN: A simultaneous Ferriman Gallwey map scoring design was used. SETTING: The study was performed at the General Clinical Research Center in a university medical center. PATIENTS: Twenty-one hirsute patients with the polycystic ovary syndrome were studied. INTERVENTIONS: Interventions included masked patient, physician, and research nurse simultaneous and independent Ferriman Gallwey map scoring. MAIN OUTCOME MEASURES: The main outcome measures were modified Ferriman Gallwey map scores. RESULTS: Agreement analysis demonstrated that patient self scores, physician scores, and research nurse scores were quite discrepant. CONCLUSION: The modified Ferriman Gallwey Map scoring system has too much variation to be clinically useful. PMID- 15827103 TI - Effects of calcium supplementation on body weight and blood pressure in normal older women: a randomized controlled trial. AB - CONTEXT: Epidemiological data suggest that high calcium intakes are associated with decreased body weight and blood pressure. However, there is little evidence from randomized trials that addresses these important issues. OBJECTIVE: The objective of this study was to assess the long-term effects of calcium on body weight and blood pressure. DESIGN: This is a substudy of an ongoing, double blind, randomized, controlled trial of calcium supplementation. End points were assessed at 30 months. SETTING: This study was performed at a university medical center. PARTICIPANTS: Normal postmenopausal women (mean age, 74 yr; mean weight, 67 kg; mean blood pressure, 134/70 mm Hg at baseline) participated in this study. INTERVENTION: Study subjects were treated with calcium (1 g/d; n = 732) and placebo (n = 739). MAIN OUTCOME MEASURES: Body weight and blood pressure were the main outcome measures. RESULTS: Weight decreased by 368 +/- 132 g (mean +/- se) with calcium treatment and by 369 +/- 134 g with placebo (P = 0.93). Fat and lean masses did not show an effect of calcium. Blood pressure showed transient reductions of 1-2 mm Hg at 6 months in the calcium group, resulting in a significant between-group difference only for systolic pressure (P = 0.048). At 30 months, the change from baseline in systolic pressure was 0.0 +/- 0.9 mm Hg in the calcium group and 2.4 +/- 0.9 mm Hg in the placebo group (P = 0.14). For diastolic pressures, the changes were -0.2 +/- 0.4 and 0.8 +/- 0.4 mm Hg, respectively (P = 0.13). In those with baseline calcium intakes less than 600 mg/d, the treatment effect was greater and did persist. CONCLUSIONS: Calcium supplementation of 1 g/d does not produce biologically significant effects on body weight, and its hypotensive effect is small and transient in most women. PMID- 15827104 TI - Single-dose pharmacokinetics and tolerability of alendronate 35- and 70-milligram tablets in children and adolescents with osteogenesis imperfecta type I. AB - CONTEXT: Alendronate (ALN) is a bisphosphonate compound that can be administered orally and has potential use in pediatric osteoporotic conditions. OBJECTIVE: The objective was to evaluate the pharmacokinetics and single-dose tolerability of ALN in children with osteogenesis imperfecta. DESIGN: ALN was administered iv and orally in a two-period, randomized crossover study, with doses separated by a 2 wk washout and follow-up carried out within 2 wk after the last ALN dose. SETTING: The study was conducted at the pediatric metabolic bone research unit at the Shriners Hospital for Children, Montreal, Canada. PATIENTS: Twenty-four children (aged 4-16 yr; eight girls) with osteogenesis imperfecta type I participated. INTERVENTIONS: All patients received iv ALN at a dose of 125 mug. In addition, patients weighing less than 40 kg received an oral dose of ALN 35 mg, whereas those weighing 40 kg or more received ALN 70 mg orally. MAIN OUTCOME MEASURES: Total urinary excretion and oral bioavailability of ALN, blood and urine safety parameters, and adverse events were the main outcome measures. RESULTS: The total urinary excretion of ALN after the iv dose was similar for both weight groups. The mean oral bioavailability (95% confidence interval) was 0.43% (0.28, 0.64%) for patients weighing less than 40 kg and 0.56% (0.36, 0.87%) for patients weighing 40 kg or more. Eighteen patients reported a total of 44 clinical adverse experiences, none of which were serious. The most common adverse experiences were mild to moderate headache (n = 7), nausea (n = 7), fever (n = 5), and abdominal pain (n = 6). Eighty percent of the adverse experiences (35 of 44) occurred within 48 h of medication administration, 91% (40 of 44) lasted less than 24 h, and 84% (37 of 44) were reported after oral dosing. Laboratory safety monitoring revealed a marginal decrease in absolute lymphocyte count and serum alkaline phosphatase after the study compared with baseline for both weight categories. CONCLUSIONS: The mean oral bioavailability of 35- and 70-mg ALN tablets was less than 0.6%, comparable to adult studies. Adverse experiences from single-dose ALN were minor, and the drug was generally well-tolerated. PMID- 15827105 TI - Effect of somatostatin on duodenal glucose absorption in man. AB - OBJECTIVE: The hyperglycemic hyperinsulinemic clamp technique using intraduodenally infused glucose is an attractive tool for studying postprandial glucose metabolism under strictly controlled conditions. Because it requires the use of somatostatin (SST), we examined, in this study, the effect of SST on intestinal glucose absorption. CONTEXT: Twenty-six normal volunteers were given a constant 3-h intraduodenal infusion of glucose (6 mg.kg(-1).min(-1)) labeled with [2-(3)H]glucose for glucose absorption measurement. During glucose infusion, 19 subjects received iv SST at doses of 10-100 ng.kg(-1).min(-1) plus insulin and glucagon, and seven subjects were studied under control conditions. In the controls, glucose was absorbed at a rate that, after a 20-min lag period, equaled the infusion rate. RESULTS: With all the doses of SST tested, absorption was considerably delayed but equaled the rate of infusion after 3 h. At that time, only 5 +/- 2% of the total amount of infused glucose was unabsorbed in the control subjects vs. 36 +/- 2% (P < 0.001) in the SST-infused subjects. In the latter, the intraluminal residue was almost totally absorbed within 40 min of the cessation of SST infusion. At the lowest dose of SST tested (10 ng.kg(-1).min( 1)), suppression of insulin secretion was incomplete. CONCLUSION: These properties of SST hamper the use of intraduodenal hyperglycemic hyperinsulinemic clamps as a tool for exploring postprandial glucose metabolism. PMID- 15827106 TI - A study of the hexose-6-phosphate dehydrogenase gene R453Q and 11beta hydroxysteroid dehydrogenase type 1 gene 83557insA polymorphisms in the polycystic ovary syndrome. AB - CONTEXT: The R453Q variant in the hexose-6-phosphate dehydrogenase gene (H6PD) and 83557insA mutations in 11beta-hydroxysteroid dehydrogenase (11betaHSD) type 1 gene (HSD11B1) interact, resulting in cortisone reductase deficiency (CRD), a rare disorder characterized by a polycystic ovary syndrome (PCOS)-like phenotype. OBJECTIVE: The objective was to study these mutations in PCOS. DESIGN: The design was a case-control study. SETTING: The study was conducted in an academic hospital. PARTICIPANTS: A total of 116 PCOS patients and 76 nonhyperandrogenic controls participated. MAIN OUTCOME MEASURES: Genotype distributions and influence of genotypes on clinical and biochemical variables and, in 28 patients and 12 controls, estimates of 11betaHSD oxoreductase activity were the main outcome measures. RESULTS: Four controls and five patients presented three of four mutant alleles in H6PD R453Q and HSD11B1 83557insA, which is the genotype observed in some subjects with CRD. Estimates of 11betaHSD oxoreductase activity were measured in six of these nine women, ruling out CRD. Moreover, H6PD R453Q and HSD11B1 83557insA genotypes, either separately or in combination, did not influence 11betaHSD oxoreductase activity. The distribution of H6PD R453Q genotypes (R/R, R/Q, and Q/Q) was different in patients and controls (42% of controls and 63% of PCOS patients were R/R; 53% of controls and 31% of PCOS patients were R/Q; and 5% of controls and 6% of PCOS patients were Q/Q; chi(2) = 9.1; P = 0.011). Patients homozygous for R453 alleles presented with increased cortisol and 17-hydroxyprogesterone levels, compared with carriers of Q453 alleles, but these differences were not observed in controls. On the contrary, HSD11B1 83557insA genotypes were not associated with PCOS and did not influence any phenotypic variable. CONCLUSIONS: Digenic triallelic genotypes of the H6PD R453Q variant and HSD11B1 83557insA mutation do not always cause CRD. On the contrary, the H6PD R453Q variant is associated with PCOS and might influence its phenotype by influencing adrenal activity. PMID- 15827107 TI - Independent and combined effects of testosterone and growth hormone on extracellular water in hypopituitary men. AB - CONTEXT: Symptoms of fluid retention in GH-deficient patients during GH replacement are greater in men than in women, suggesting that testosterone may augment or estradiol may attenuate the antinatriuretic actions of GH. The mechanisms underlying the sodium-retaining effects of GH are poorly understood. AIM: The aim of this study was to investigate the effects of GH and testosterone, alone and in combination, on extracellular water (ECW) and the hormonal mechanisms involved. DESIGN: Two separate, open-label, randomized, two-period, crossover studies were performed; the first compared the effects of GH alone with those of GH and testosterone, and the second compared the effects of testosterone alone with those of GH and testosterone. PARTICIPANTS: Twelve hypopituitary men with GH deficiency and hypogonadism were studied. INTERVENTION: During the weeks of intervention, GH (0.5 mg/d) and testosterone enanthate (250 mg) were administered by im injection. OUTCOME MEASURES: The outcome measures were ECW, IGF-I, plasma renin activity (PRA), aldosterone (Aldo), and atrial natriuretic peptide (ANP). RESULTS: GH treatment significantly increased (P < 0.05) both IGF I and ECW, and these changes were enhanced by cotreatment with testosterone (P = 0.07 for both). PRA, Aldo, and ANP levels did not change. Testosterone treatment alone did not change the IGF-I concentration, whereas cotreatment with GH induced a marked increase. Testosterone alone increased (P < 0.05) ECW, and the effect was augmented (P < 0.01) by cotreatment with GH. Although PRA and ANP did not change, plasma Aldo decreased after single and combined treatments. CONCLUSION: GH and testosterone exerted independent and additive effects on ECW. The mechanisms of fluid retention for both hormones are likely to be exerted on the renal tubules. This is the first direct evidence that testosterone increases ECW. PMID- 15827108 TI - Fibroblast growth factor-23 in patients with Graves' disease before and after antithyroid therapy: its important role in serum phosphate regulation. AB - OBJECTIVE: Hyperthyroidism is a well-described cause of hyperphosphatemia. We aimed to clarify the physiological role of fibroblast growth factor (FGF)-23 in serum phosphate homeostasis in patients with Graves' disease during the course of treatment for hyperthyroidism. CONTEXT: The study group comprised 56 patients (45 for a cross-sectional study and 11 for a longitudinal study) with Graves' disease. For the cross-sectional study, patients were assigned, on the basis of their serum phosphate level, to a hypophosphatemia group (n = 14), a normophosphatemia group (n = 16), or a hyperphosphatemia group (n = 15). Serum FGF-23, calcium, phosphate, PTH, and 1,25-dihydroxyvitamin D [1,25(OH)(2)D] levels were compared between the three groups. For the longitudinal study, we assessed changes in these biochemical indices before and after antithyroid treatment. RESULTS: In the cross-sectional study, the serum FGF-23 level was significantly higher (P < 0.05) in the hyperphosphatemia group than in the other groups (61 +/- 36 ng/liter vs. 31 +/- 22 ng/liter and 30 +/- 9 ng/liter). In the longitudinal study, serum levels of FGF-23 decreased significantly (P < 0.05) from a high of 54 +/- 12 ng/liter before treatment to 29 +/- 14 ng/liter after treatment. In contrast, the serum 1,25(OH)(2)D level increased significantly (P < 0.005) from 55 +/- 22 pmol/liter before treatment to 185 +/- 76 pmol/liter 3 months after treatment. Serum FGF-23 levels were positively correlated with serum phosphate levels (P < 0.0001) and negatively correlated with serum 1,25(OH)(2)D levels (P < 0.0001). CONCLUSIONS: The significant positive correlation between serum levels of phosphate and FGF-23 indicates that FGF-23 may play an important role in serum phosphate homeostasis by its up-regulation in the hyperphosphatemic condition. PMID- 15827109 TI - A critical analysis of pituitary tumor shrinkage during primary medical therapy in acromegaly. AB - CONTEXT: Somatostatin analogs have been successfully used to treat patients with GH-secreting pituitary adenomas because they are safe, effective, and usually well tolerated. The results of studies evaluating acromegaly treatment with the somatostatin receptor ligands (SRLs), octreotide and lanreotide, have supported the use of these agents for primary medical therapy before or as an alternative to traditional interventions of surgery and radiotherapy in selected cases. EVIDENCE ACQUISITION: We therefore undertook a systematic literature overview to characterize the results of studies involving primary therapy with somatostatin analogs and their effects on pituitary tumor size. Because most studies in which pituitary tumor shrinkage has been assessed involve uncontrolled, open-label, prospective trials or retrospective case series, the lack of a control arm does not permit pooling of data in a metaanalytic fashion to determine tumor size reduction. Therefore, this systematic review was designed to document and stratify data by study design, summarize therapeutic regimens and patient characteristics, assess the percentage of patients showing changes in tumor size, and calculate the weighted average effect on size reduction. EVIDENCE SYNTHESIS: Overall, for patients who experience significant shrinkage, an approximately 50% decrease in pituitary mass is achieved when a somatostatin analog is used exclusively or before surgery or radiotherapy. Fourteen studies (n = 424) provided a definition of significant tumor shrinkage, and the results showed that 36.6% (weighted mean percentage) of patients receiving primary SRL therapy for acromegaly experienced a significant reduction in tumor size. The weighted mean percent reduction in tumor size was 19.4% for those studies in which all patients received SRLs and change in tumor size was reported for all patients. CONCLUSIONS: Clinical implications are discussed for patients in whom tumor size control with SRLs is an important objective, typically those who have failed surgery or are being treated with primary medical therapy with large tumors. PMID- 15827110 TI - The hypothalamic-pituitary-adrenal axis in pregnancy: challenges in disease detection and treatment. AB - Pregnancy dramatically affects the hypothalamic-pituitary-adrenal axis leading to increased circulating cortisol and ACTH levels during gestation, reaching values in the range seen in Cushing's syndrome (CS). The cause(s) of increased ACTH may include placental synthesis and release of biologically active CRH and ACTH, pituitary desensitization to cortisol feedback, or enhanced pituitary responses to corticotropin-releasing factors. In this context, challenges in diagnosis and management of disorders of the hypothalamic-pituitary-adrenal axis in pregnancy are discussed. CS in pregnancy is uncommon and is associated with fetal morbidity and mortality. The diagnosis may be missed because of overlapping clinical and biochemical features in pregnancy. The proportion of patients with primary adrenal causes of CS is increased in pregnancy. CRH stimulation testing and inferior petrosal sinus sampling can identify patients with Cushing's disease. Surgery is a safe option for treatment in the second trimester; otherwise medical therapy may be used. Women with known adrenal insufficiency that is appropriately treated can expect to have uneventful pregnancies. Whereas a fetal/placental source of cortisol may mitigate crisis during gestation, unrecognized adrenal insufficiency may lead to maternal or fetal demise either during gestation or in the puerperium. Appropriate treatment and management of labor are reviewed. PMID- 15827111 TI - Insight into Graves' hyperthyroidism from animal models. AB - Graves' hyperthyroidism can be induced in mice or hamsters by novel approaches, namely injecting cells expressing the TSH receptor (TSHR) or vaccination with TSHR-DNA in plasmid or adenoviral vectors. These models provide unique insight into several aspects of Graves' disease: 1) manipulating immunity toward Th1 or Th2 cytokines enhances or suppresses hyperthyroidism in different models, perhaps reflecting human disease heterogeneity; 2) the role of TSHR cleavage and A subunit shedding in immunity leading to thyroid-stimulating antibodies (TSAbs); and 3) epitope spreading away from TSAbs and toward TSH-blocking antibodies in association with increased TSHR antibody titers (as in rare hypothyroid patients). Major developments from the models include the isolation of high affinity monoclonal TSAbs and analysis of antigen presentation, T cells, and immune tolerance to the TSHR. Studies of inbred mouse strains emphasize the contribution of non-MHC vs. MHC genes, as in humans, supporting the relevance of the models to human disease. Moreover, other findings suggest that the development of Graves' disease is affected by environmental factors, including infectious pathogens, regardless of modifications in the Th1/Th2 balance. Finally, developing immunospecific forms of therapy for Graves' disease will require painstaking dissection of immune recognition and responses to the TSHR. PMID- 15827112 TI - Architectural and synaptic mechanisms underlying coherent spontaneous activity in V1. AB - To investigate the existence and the characteristics of possible cortical operating points of the primary visual cortex, as manifested by the coherent spontaneous ongoing activity revealed by real-time optical imaging based on voltage-sensitive dyes, we studied numerically a very large-scale ( approximately 5 x 10(5)) conductance-based, integrate-and-fire neuronal network model of an approximately 16-mm(2) patch of 64 orientation hypercolumns, which incorporates both isotropic local couplings and lateral orientation-specific long-range connections with a slow NMDA component. A dynamic scenario of an intermittent desuppressed state (IDS) is identified in the computational model, which is a dynamic state of (i) high conductance, (ii) strong inhibition, and (iii) large fluctuations that arise from intermittent spiking events that are strongly correlated in time as well as in orientation domains, with the correlation time of the fluctuations controlled by the NMDA decay time scale. Our simulation results demonstrate that the IDS state captures numerically many aspects of experimental observation related to spontaneous ongoing activity, and the specific network mechanism of the IDS may suggest cortical mechanisms and the cortical operating point underlying observed spontaneous activity. PMID- 15827113 TI - Coupling neurotrophins to cell migration through selective guanine nucleotide exchange factor activation. PMID- 15827114 TI - Viral trafficking violations in axons: the herpesvirus case. PMID- 15827115 TI - Organization of synaptic myonuclei by Syne proteins and their role during the formation of the nerve-muscle synapse. PMID- 15827116 TI - Late-onset motoneuron disease caused by a functionally modified AMPA receptor subunit. AB - Amyotrophic lateral sclerosis (ALS) is a devastating disorder of the central nervous system in middle and old age that leads to progressive loss of spinal motoneurons. Transgenic mice overexpressing mutated human Cu(2+)/Zn(2+) superoxide dismutase 1 (SOD1) reproduce clinical features of the familial form of ALS. However, changes in SOD1 activity do not correlate with severity of motor decline in sporadic cases, indicating that targets unrelated to superoxide metabolism contribute to the pathogenesis of the disease. We show here that transgenic expression in mice of GluR-B(N)-containing L-alpha-amino-3-hydroxy-5 methylisoxazole-4-proprionate (AMPA) receptors with increased Ca(2+) permeability leads to late-onset degeneration of neurons in the spinal cord and decline of motor functions. Neuronal death progresses over the entire lifespan but manifests clinically in late adulthood, resembling the course of a slow neurodegenerative disorder. Additional transgenic expression of mutated human SOD1 accelerates disease progression, aggravates the severity of motor decline, and decreases survival. These observations link persistently elevated Ca(2+) influx through AMPA channels with progressive motor decline and late-onset degeneration of spinal motoneurons, indicating that functionally altered AMPA channels may be causally related to pathogenesis of sporadic ALS in humans. PMID- 15827117 TI - Fluorescence measurements reveal stoichiometry of K+ channels formed by modulatory and delayed rectifier alpha-subunits. AB - Modulatory alpha-subunits, which comprise one-fourth of all voltagegated K(+) channel (Kv) alpha-subunits, do not assemble into homomeric channels, but selectively associate with delayed rectifier Kv2 subunits to form heteromeric channels of unknown stoichiometry. Their distinct expression patterns and unique functional properties have made these channels candidate molecular correlates for a broad set of native K(+) currents. Here, we combine FRET and electrophysiological measurements to determine the stoichiometry and geometry of heteromeric channels composed of the delayed rectifier Kv2.1 subunit and the modulatory Kv9.3 alpha-subunit. Kv channel alpha-subunits were fused with GFP variants, and heteromerization of different combinations of tagged and untagged alpha-subunits was studied. FRET, evaluated by acceptor photobleaching, was only observed upon formation of functional channels. Our results, obtained from two independent experimental paradigms, suggest the formation of heteromeric Kv2.1/Kv9.3 channels of fixed stoichiometry consisting of three Kv2.1 subunits and one Kv9.3 subunit. Strikingly, despite this uneven stoichiometry, we find that heteromeric Kv2.1/Kv9.3 channels maintain a pseudosymmetric arrangement of subunits around the central pore. PMID- 15827118 TI - Generalizations of the classical Weyl and Colin de Verdiere's formulas and the orbit method. AB - The classical Weyl formula expresses the leading term of the asymptotics of the counting function N(lambda, H) of the spectrum of a self-adjoint operator H in an invariant form: one can "hear" the volume of the subset of the cotangent bundle where the symbol of the operator H is less than lambda. In particular, it is applicable to Schrodinger operators with electric potentials growing at infinity. The Weyl formula is formulated in an invariant form; however, it gives +infinity for magnetic Schrodinger operators with magnetic tensors growing at infinity. For these operators, Colin de Verdiere's formula is known, but the form of the latter is not invariant. In this article, we suggest an invariant generalization of both Weyl's and Colin de Verdiere's formulas for wide classes of Schrodinger operators with polynomial electric and magnetic fields. The construction is based on the orbit method due to Kirillov, and it allows one to hear the geometry of coadjoint orbits. PMID- 15827119 TI - Rates of intron loss and gain: implications for early eukaryotic evolution. AB - We study the intron-exon structures of 684 groups of orthologs from seven diverse eukaryotic genomes and provide maximum likelihood estimates for rates and numbers of intron losses and gains in these same genes for a variety of lineages. Rates of intron loss vary from approximately 2 x 10(-9) to 2 x 10(-10) per year. Rates of gain vary from 6 x 10(-13) to 4 x 10(-12) per possible intron insertion site per year. There is an inverse correspondence between rates of intron loss and gain, leading to a 20-fold variation among lineages in the ratio of the rates of the two processes. The observed rates of intron gain are insufficient to explain the large number of introns estimated to have been present in the plant-animal ancestor, suggesting that introns present in early eukaryotes may have been created by a fundamentally different process than more recently gained introns. PMID- 15827120 TI - A glimpse at the organization of the protein universe. PMID- 15827121 TI - Cre/lox-regulated transgenic zebrafish model with conditional myc-induced T cell acute lymphoblastic leukemia. AB - We have created a stable transgenic rag2-EGFP-mMyc zebrafish line that develops GFP-labeled T cell acute lymphoblastic leukemia (T-ALL), allowing visualization of the onset and spread of this disease. Here, we show that leukemias from this transgenic line are highly penetrant and render animals moribund by 80.7 +/- 17.6 days of life (+/-1 SD, range = 50-158 days). These T cell leukemias are clonally aneuploid, can be transplanted into irradiated recipient fish, and express the zebrafish orthologues of the human T-ALL oncogenes tal1/scl and lmo2, thus providing an animal model for the most prevalent molecular subgroup of human T ALL. Because T-ALL develops very rapidly in rag2-EGFP-mMyc transgenic fish (in which "mMyc" represents mouse c-Myc), this line can only be maintained by in vitro fertilization. Thus, we have created a conditional transgene in which the EGFP-mMyc oncogene is preceded by a loxed dsRED2 gene and have generated stable rag2-loxP-dsRED2-loxP-EGFP-mMyc transgenic zebrafish lines, which have red fluorescent thymocytes and do not develop leukemia. Transgenic progeny from one of these lines can be induced to develop T-ALL by injecting Cre RNA into one-cell stage embryos, demonstrating the utility of the Cre/lox system in the zebrafish and providing an essential step in preparing this model for chemical and genetic screens designed to identify modifiers of Myc-induced T-ALL. PMID- 15827122 TI - Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. AB - Biomaterial surface chemistry has profound consequences on cellular and host responses, but the underlying molecular mechanisms remain poorly understood. Using self-assembled monolayers as model biomaterial surfaces presenting well defined chemistries, we demonstrate that surface chemistry modulates osteoblastic differentiation and matrix mineralization independently from alterations in cell proliferation. Surfaces were precoated with equal densities of fibronectin (FN), and surface chemistry modulated FN structure to alter integrin adhesion receptor binding. OH- and NH(2)-terminated surfaces up-regulated osteoblast-specific gene expression, alkaline phosphatase enzymatic activity, and matrix mineralization compared with surfaces presenting COOH and CH(3) groups. These surface chemistry dependent differences in cell differentiation were controlled by binding of specific integrins to adsorbed FN. Function-perturbing antibodies against the central cell binding domain of FN completely inhibited matrix mineralization. Furthermore, blocking antibodies against beta(1) integrin inhibited matrix mineralization on the OH and NH(2) surfaces, whereas function-perturbing antibodies specific for beta(3) integrin increased mineralization on the COOH substrate. These results establish surface-dependent differences in integrin binding as a mechanism regulating differential cellular responses to biomaterial surfaces. This mechanism could be exploited to engineer materials that control integrin binding specificity to elicit desired cellular activities to enhance the integration of biomaterials and improve the performance of biotechnological culture supports. PMID- 15827123 TI - Gene expression profiling reveals molecularly and clinically distinct subtypes of glioblastoma multiforme. AB - Glioblastoma multiforme (GBM) is the most common form of malignant glioma, characterized by genetic instability, intratumoral histopathological variability, and unpredictable clinical behavior. We investigated global gene expression in surgical samples of brain tumors. Gene expression profiling revealed large differences between normal brain samples and tumor tissues and between GBMs and lower-grade oligodendroglial tumors. Extensive differences in gene expression were found among GBMs, particularly in genes involved in angiogenesis, immune cell infiltration, and extracellular matrix remodeling. We found that the gene expression patterns in paired specimens from the same GBM invariably were more closely related to each other than to any other tumor, even when the paired specimens had strikingly divergent histologies. Survival analyses revealed a set of approximately 70 genes more highly expressed in rapidly progressing tumors that stratified GBMs into two groups that differed by >4-fold in median duration of survival. We further investigated one gene from the group, FABP7, and confirmed its association with survival in two unrelated cohorts totaling 105 patients. Expression of FABP7 enhanced the motility of glioma-derived cells in vitro. Our analyses thus identify and validate a prognostic marker of both biologic and clinical significance and provide a series of putative markers for additional evaluation. PMID- 15827124 TI - A population-epigenetic model to infer site-specific methylation rates from double-stranded DNA methylation patterns. AB - Cytosine methylation is an epigenetic mechanism in eukaryotes that is often associated with stable transcriptional silencing, such as in X-chromosome inactivation and genomic imprinting. Aberrant methylation patterns occur in several inherited human diseases and in many cancers. To understand how methylated and unmethylated states of cytosine residues are transmitted during DNA replication, we develop a population-epigenetic model of DNA methylation dynamics. The model is informed by our observation that de novo methylation can occur on the daughter strand while leaving the opposing cytosine unmethylated, as revealed by the patterns of methylation on the two complementary strands of individual DNA molecules. Under our model, we can infer site-specific rates of both maintenance and de novo methylation, values that determine the fidelity of methylation inheritance, from double-stranded methylation data. This approach can be used for populations of cells obtained from individuals without the need for cell culture. We use our method to infer cytosine methylation rates at several sites within the promoter of the human gene FMR1. PMID- 15827125 TI - Mutant-specific gene programs in the zebrafish. AB - Hematopoiesis involves the production of stem cells, followed by the orchestrated differentiation of the blood lineages. Genetic screens in zebrafish have identified mutants with defects that disrupt specific stages of hematopoiesis and vasculogenesis, including the cloche, spadetail (tbx16), moonshine (tif1g), bloodless, and vlad tepes (gata1) mutants. To better characterize the blood program, gene expression profiling was carried out in these mutants and in scl morphants (scl(mo)). Distinct gene clusters were demarcated by stage-specific and mutant-specific gene regulation. These were found to correlate with the transcriptional program of hematopoietic progenitor cells, as well as of the erythroid, myeloid, and vascular lineages. Among these, several novel hematopoietic and vascular genes were detected, for instance, the erythroid transcription factors znfl2 and ncoa4. A specific regulation was found for myeloid genes, as they were more strongly expressed in vlt mutants compared with other erythroid mutants. A unique gene expression pattern of up-regulated isoprenoid synthesis genes was found in cloche and scl(mo), possibly in migrating cells. In conjunction with the high conservation of vertebrate hematopoiesis, the comparison of transcriptional profiles in zebrafish blood mutants represents a versatile and powerful tool to elucidate the genetic regulation of blood and blood vessel development. PMID- 15827126 TI - Siglec-9 transduces apoptotic and nonapoptotic death signals into neutrophils depending on the proinflammatory cytokine environment. AB - We report about new apoptotic and non-apoptotic death pathways in neutrophils that are initiated via the surface molecule sialic acid-binding immunoglobulin like lectin (Siglec)-9. In normal neutrophils, Siglec-9 ligation induced apoptosis. Inflammatory neutrophils obtained from patients with acute septic shock or rheumatoid arthritis demonstrated increased Siglec-9, but normal Fas receptor-mediated cytotoxic responses when compared with normal blood neutrophils. The increased Siglec-9-mediated death was mimicked in vitro by short term preincubation of normal neutrophils with proinflammatory cytokines, such as granulocyte/macrophage colony-stimulating factor (GM-CSF), interferon-alpha (IFN alpha), and IFN-gamma, and was demonstrated to be caspase independent. Experiments using scavengers of reactive oxygen species (ROS) or neutrophils unable to generate ROS indicated that both Siglec-9-mediated caspase-dependent and caspase-independent forms of neutrophil death depend on ROS. Interestingly, the caspase-independent form of neutrophil death was characterized by cytoplasmic vacuolization and several other nonapoptotic morphologic features, which were also seen in neutrophils present in joint fluids from rheumatoid arthritis patients. Taken together, these data suggest that apoptotic (ROS- and caspase dependent) and nonapoptotic (ROS-dependent) death pathways are initiated in neutrophils via Siglec-9. The new insights have important implications for the pathogenesis, diagnosis, and treatment of inflammatory diseases such as sepsis and rheumatoid arthritis. PMID- 15827127 TI - Preferential suppression of trisomy 8 compared with normal hematopoietic cell growth by autologous lymphocytes in patients with trisomy 8 myelodysplastic syndrome. AB - Clinical observations and experimental evidence link bone marrow failure in myelodysplastic syndrome (MDS) with a T cell-dominated autoimmune process. Immunosuppressive therapy is effective in improving cytopenias in selected patients. Trisomy 8 is a frequent cytogenetic abnormality in bone marrow cells in patients with MDS, and its presence has been associated anecdotally with good response to immunotherapy. We studied 34 patients with trisomy 8 in bone marrow cells, some of whom were undergoing treatment with antithymocyte globulin (ATG). All had significant CD8+ T-cell expansions of one or more T-cell receptor (TCR) Vbeta subfamilies, as measured by flow cytometry; expanded subfamilies showed CDR3 skewing by spectratyping. Sorted T cells of the expanded Vbeta subfamilies, but not of the remaining subfamilies, inhibited trisomy 8 cell growth in short term hematopoietic culture. The negative effects of Vbeta-expanded T cells were inhibited by major histocompatibility complex (MHC) class 1 monoclonal antibody (mAb) and Fas antagonist and required direct cell-to-cell contact. Sixty-seven percent of patients who had de novo MDS with trisomy 8 as the sole karyotypic abnormality responded to ATG with durable reversal of cytopenias and restoration of transfusion independence, with stable increase in the proportion of trisomy 8 bone marrow cells and normalization of the T-cell repertoire. An increased number of T cells with apparent specificity for trisomy 8 cells is consistent with an autoimmune pathophysiology in trisomy 8 MDS. PMID- 15827128 TI - Seliciclib (CYC202 or R-roscovitine), a small-molecule cyclin-dependent kinase inhibitor, mediates activity via down-regulation of Mcl-1 in multiple myeloma. AB - Cyclin-dependent kinase (CDK) inhibitors have the potential to induce cell-cycle arrest and apoptosis in cancer cells. Seliciclib (CYC202 or R-roscovitine) is a potent CDK inhibitor currently undergoing phase-2 clinical testing in lung and B cell malignancies. Here we studied the in vitro cytotoxic activity of seliciclib against multiple myeloma (MM) cells. Our data demonstrate that seliciclib has potent cytotoxicity against MM cells that are both sensitive and resistant to conventional therapy as well as primary MM cells from patients. Cell-cycle and Western blot analysis confirmed apoptosis. Importantly, seliciclib triggered a rapid down-regulation of Mcl-1 transcription and protein expression independent of caspase cleavage. Adherence of MM cells to bone marrow stromal cells (BMSCs) induced increased Mcl-1 expression associated with signal transducer and activator of transcription 3 (STAT3) phosphorylation, which was inhibited in a time- and dose-dependent manner by seliciclib. Furthermore, seliciclib inhibited interleukin 6 (IL-6) transcription and secretion triggered by tumor cell binding to BMSCs. Up-regulation of Mcl-1 expression in cocultures was only partially blocked by neutralizing antibody to IL-6, suggesting alternative mechanisms of Mcl-1 modulation by seliciclib. Finally, combination studies of seliciclib with doxorubicin and bortezomib show in vitro synergism, providing the rationale for testing these drug combinations to improve patient outcome in MM. PMID- 15827129 TI - Rituximab prevents recurrence of thrombotic thrombocytopenic purpura: a case report. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare disorder of small vessels that is associated with deficiency of the von Willebrand factor-cleaving protease, ADAMTS13. The presence of anti-ADAMTS13 autoantibodies is considered a factor predisposing to relapses. Despite close monitoring and intensive plasma treatment, in these patients acute episodes are still associated with substantial morbidity and mortality rates, and the optimal therapeutic option should be prevention of relapses. This study was conducted in a patient with recurrent TTP due to high titers of ADAMTS13 inhibitors, who used to have 2 relapses of TTP a year. The study compared the standard treatment plasma exchange with rituximab. Results documented that plasma exchange had only a small transient effect on ADAMTS13 activity and inhibitors; on the contrary, prophylaxis with rituximab was associated with disappearance of anti-ADAMTS13 antibodies, a progressive recovery of protease activity, and it allowed the patient to maintain a disease-free state during a more than 2-year follow-up. PMID- 15827130 TI - Identification and characterization of epitopes of the receptor for hyaluronic acid-mediated motility (RHAMM/CD168) recognized by CD8+ T cells of HLA-A2 positive patients with acute myeloid leukemia. AB - The receptor for hyaluronic acid-mediated motility (RHAMM/CD168) has been described as a leukemia-associated antigen. To define T-cell epitopes of RHAMM/CD168 toward specific immunotherapies for acute myeloid leukemia (AML), 10 potential HLA-A2-binding RHAMM/CD168 peptides (R1 to R10) were synthesized based on computer algorithms and screened by enzyme-linked immunospot (ELISPOT) analysis using CD8+ T cells isolated from peripheral blood (PB) of patients with AML and healthy donors. We found that CD8+ cells from 7 of 13 (54%) patients with AML presensitized with peptides R3 (ILSLELMKL) or R5 (SLEENIVIL) specifically recognized T2 cells pulsed with R3 (39%) or R5 (15%) peptide. In contrast, only 4 of 21 (19%) healthy volunteers had CD8+ cells reactive with R3- or R5-pulsed T2 cells after presensitization. The presence of R3 peptide-specific effector T cells in the peripheral blood of patients with AML could be confirmed by staining as HLA-A2/R3 peptide tetramer+ CCR7-CD45RA+ cells. In chromium-51 release assays, peptide-primed CD8+ T cells from patients with AML were able to lyse RHAMM/CD168 peptide-pulsed T2 cells, AML blasts, and dendritic cells generated thereof (AML DCs). Transfection of COS7 cells with RHAMM/CD168 cDNA revealed that peptides R3 and R5 are naturally processed epitopes of RHAMM/CD168 that are presented in an HLA-A2-restricted manner. In summary, RHAMM/CD168 is a promising target for immunotherapies in patients with AML, and we have therefore initiated a clinical vaccination trial with R3 peptide. Because RHAMM/CD168 is also expressed in various other hematologic malignancies and solid tumors, vaccines targeting this antigen may have even wider application. PMID- 15827131 TI - RNA interference in biology and disease. AB - RNA interference (RNAi) is a conserved biologic response to double-stranded RNA that results in the sequence-specific silencing of target gene expression. Over the past 5 years, an intensive research effort has facilitated the rapid movement of RNAi from a relatively obscure biologic phenomenon to a valuable tool used to silence target gene expression and perform large-scale functional genomic screens. In fact, recent studies reported in this journal and others have demonstrated success using RNAi to address the role of oncogene expression in leukemia cell lines and to validate the therapeutic potential of RNAi for treating these blood disorders. In order to advance these applications and gain an appreciation for the future of RNAi both in basic research and in the treatment of diseases caused by aberrant gene expression, it is important to have an understanding of the process of RNAi and its limitations. PMID- 15827132 TI - Conservation of unique cell-surface CD antigen mosaics in HIV-1-infected individuals. AB - Cluster of differentiation (CD) antigens are expressed on cells of myeloid and lymphoid lineages. As most disease processes involve immune system activation or suppression, these antigens offer unique opportunities for monitoring host responses. Immunophenotyping using limited numbers of CD antigens enables differentiation states of immune system cells to be determined. Extended phenotyping involving parallel measurement of multiple CD antigens may help identify expression pattern signatures associated with specific disease states. To explore this possibility we have made a CD monoclonal antibody array and scanner, enabling the parallel immunophenotyping of leukocyte cell suspensions in a single and rapid analysis. To demonstrate this approach, we used the specific example of patients infected with human immunodeficiency virus type-1 (HIV-1). An invariant HIV-induced CD antigen signature has been defined that is both robust and independent of clinical outcome, composed of a unique profile of CD antigen expression levels that are both increased and decreased relative to internal controls. The results indicate that HIV-induced changes in CD antigen expression are disease specific and independent of outcome. Their invariant nature indicates an irreversible component to retroviral infection and suggests the utility of CD antigen expression patterns in other disease settings. PMID- 15827133 TI - Regional induction of adhesion molecules and chemokine receptors explains disparate homing of human B cells to systemic and mucosal effector sites: dispersion from tonsils. AB - Ethical constraints restrict direct tracking of immune-cell migration throughout the human body in vivo. We, therefore, used deletion of the immunoglobulin M (IgM) heavy-chain constant-gene (Cmu) segment as a marker to provide a dispersal signature of an effector B-cell subset (IgD(+)IgM(-)CD38+) induced selectively in human tonsils. By DNA analysis, the Cmu deletion identified dissemination of such blasts and their plasma-cell progeny to peripheral blood, lymph nodes, and bone marrow, as well as to mucosae and glands of the upper airways. Also the endocervix was often positive, while the small intestine was mainly negative, as could be expected from the identified homing-molecule profile of the marker cells, with relatively low levels of integrin alpha4beta7 and CC chemokine receptor 9 (CCR9). Of further importance for vaccine design, the circulating cells expressed abundantly CD62L (L-selectin) and CCR7, which provided a mechanism for integration of respiratory and systemic immunity. Most mucosal vaccines are at present administered perorally, and our results suggested that the nasal route is no alternative for vaccination against rotavirus or other small-intestinal infections in humans. However, immunization of nasopharynx associated lymphoid tissue clearly appears preferable to target respiratory pathogens and may to some extent also protect against infections of the female genital tract. PMID- 15827134 TI - The level of TACI gene expression in myeloma cells is associated with a signature of microenvironment dependence versus a plasmablastic signature. AB - B-cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) have been shown to promote multiple myeloma (MM) cell growth. We show that the main site of production for BAFF and APRIL is the bone marrow (BM) environment, and that production is mainly by monocytes and neutrophils. In addition, osteoclasts produce very high levels of APRIL, unlike BM stromal cells. Myeloma cells (MMCs) express TACI (transmembrane activator and calcium modulator and cyclophilin ligand interactor), the receptor of BAFF/APRIL, at varying levels. TACI expression is a good indicator of a BAFF-binding receptor. Expression data of purified MMCs from 65 newly diagnosed patients have been generated using Affymetrix microarrays and were analyzed by supervised clustering of groups with higher (TACI(hi)) versus lower (TACI(lo)) TACI expression levels. Patients in the TACI(lo) group had clinical parameters associated with bad prognosis. A set of 659 genes was differentially expressed between TACI(hi) and TACI(lo) MMCs. This set makes it possible to efficiently classify TACI(hi) and TACI(lo) MMCs in an independent cohort of 40 patients. TACI(hi) MMCs displayed a mature plasma cell gene signature, indicating dependence on the BM environment. In contrast, the TACI(lo) group had a gene signature of plasmablasts, suggesting an attenuated dependence on the BM environment. Taken together, our findings suggest using gene expression profiling to identify the group of patients who might benefit most from treatment with BAFF/APRIL inhibitors. PMID- 15827135 TI - Junctional adhesion molecule-A deficiency increases hepatic ischemia-reperfusion injury despite reduction of neutrophil transendothelial migration. AB - The endothelial receptors that control leukocyte transmigration in the postischemic liver are not identified. We investigated the role of junctional adhesion molecule-A (JAM-A), a receptor expressed in endothelial tight junctions, leukocytes, and platelets, for leukocyte transmigration during hepatic ischemia reperfusion (I/R) in vivo. We show that JAM-A is up-regulated in hepatic venular endothelium during reperfusion. I/R-induced neutrophil transmigration was attenuated in both JAM-A-/- and endothelial JAM-A-/- mice as well as in mice treated with an anti-JAM-A antibody, whereas transmigration of T cells was JAM-A independent. Postischemic leukocyte rolling remained unaffected in JAM-A-/- and endothelial JAM-A-/- mice, whereas intravascular leukocyte adherence was increased. The extent of interactions of JAM-A-/- platelets with the postischemic endothelium was comparable with that of JAM-A+/+ platelets. The I/R-induced increase in the activity of alanine aminotransferase (ALT)/aspartate aminotransferase (AST) and sinusoidal perfusion failure was not reduced in JAM-A /- mice, while the number of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL)-positive hepatocytes was significantly higher. Thus, we show for the first time that JAM-A is up-regulated in hepatic venules and serves as an endothelial receptor of neutrophil transmigration, but it does not mediate leukocyte rolling, adhesion, or platelet endothelial cell interactions. JAM-A deficiency does not reduce I/R-induced microvascular and hepatocellular necrotic injury, but increases hepatocyte apoptosis, despite attenuation of neutrophil infiltration. PMID- 15827136 TI - Effects of aging on the homing and engraftment of murine hematopoietic stem and progenitor cells. AB - To test the hypothesis that aging has negative effects on stem-cell homing and engraftment, young or old C57BL/6 bone marrow (BM) cells were injected, using a limiting-dilution, competitive transplantation method, into old or young Ly5 congenic mice. Numbers of hematopoietic stem cells (HSCs) and progenitor cells (HPCs) recovered from BM or spleen were measured and compared with the numbers initially transplanted. Although the frequency of marrow competitive repopulation units (CRUs) increased approximately 2-fold from 2 months to 2 years of age, the BM homing efficiency of old CRUs was approximately 3-fold lower than that of young CRUs. Surprisingly, the overall size of individual stem-cell clones generated in recipients receiving a single CRU was not affected by donor age. However, the increased ages of HSC donors and HSC transplant recipients caused marked skewing of the pattern of engraftment toward the myeloid lineage, indicating that HSC-intrinsic and HSC-extrinsic (microenvironmental) age-related changes favor myelopoiesis. This correlated with changes after transplantation in the rate of recovery of circulating leukocytes, erythrocytes, and platelets. Recovery of the latter was especially blunted in aged recipients. Collectively, these findings may have implications for clinical HSC transplantation in which older persons increasingly serve as donors for elderly patients. PMID- 15827137 TI - Ly49 and CD94/NKG2 receptor acquisition by NK cells does not require lymphotoxin beta receptor expression. AB - A crucial step in murine natural killer (NK) cell development, mediated by bone marrow stromal cells, is the induction of Ly49 and CD94/NKG2 receptor expression. The signals that regulate Ly49 receptor expression are still largely undetermined. It has been shown that interaction between lymphotoxin alpha1beta2 (LTalpha1beta2) and LTbeta receptor (LTbetaR), expressed on lymphoid progenitor cells and nonlymphoid bone marrow stromal cells, respectively, is important for both quantitative and functional NK cell development. Therefore, we have investigated the role of LT-LTbetaR-mediated signaling in Ly49 and CD94/NKG2 receptor acquisition. We show that the NK receptor repertoire of LTbetaR-/- mice can only be partially analyzed because of the residual 129/Ola mouse genetic background, due to a physical linkage of the LTbetaR locus and the loci encoding the Ly49 and CD94/NKG2 receptors. Therefore, we transferred wild-type B6 lymphoid committed progenitor cells into LTbetaR-/- mice, which differentiated into NK cells with a normal NK cell receptor repertoire. Also, administration of LTbetaR immunoglobulin (Ig), which acts as a soluble receptor for LTalpha1beta2, resulted in reduced NK cell percentages but did not influence the Ly49 and CD94/NKG2 receptor acquisition on remaining NK cells. These results indicate that LTbetaR mediated signals are not required for Ly49 and CD94/NKG2 receptor acquisition. PMID- 15827138 TI - Transcriptional activation of alpha/beta interferon genes: interference by nonsegmented negative-strand RNA viruses. PMID- 15827139 TI - The common viral insertion site Evi12 is located in the 5'-noncoding region of Gnn, a novel gene with enhanced expression in two subclasses of human acute myeloid leukemia. AB - The leukemia and lymphoma disease locus Evi12 was mapped to the noncoding region of a novel gene, Gnn (named for Grp94 neighboring nucleotidase), that is located immediately upstream of the Grp94/Tra1 gene on mouse chromosome 10. The Gnn gene is conserved in mice and humans. Expression of fusion constructs between GFP and Gnn cDNA isoforms in HEK-293 cells showed that Gnn proteins are located mainly in the cytoplasm. Immunoblotting experiments demonstrated the presence of multiple Gnn protein isoforms in most organs, with the lowest levels of expression of the protein detected in bone marrow and spleen. The Evi12-containing leukemia cell line NFS107 showed high levels of expression of a approximately 150-kDa Gnn isoform (Gnn107) that was not observed in control cell lines. Overexpression may be due to the viral insertion in Evi12. The Gnn107 protein is probably encoded by a Gnn cDNA isoform that is expressed exclusively in NFS107 cells and that includes sequences of TU12B1-TY, a putative protein with homology to 5' nucleotidase enzymes. Interestingly, using Affymetrix gene expression data of a cohort of 285 patients with acute myeloid leukemia (AML), we found that GNN/TU12B1-TY expression was specifically increased in two AML clusters. One cluster consisted of all AML patients with a t(8;21) translocation, and the second cluster consisted of AML patients with a normal karyotype carrying a FLT3 internal tandem duplication. These findings suggest that we identified a novel proto-oncogene that may be causally linked to certain types of human leukemia. PMID- 15827140 TI - Transmission barriers for bovine, ovine, and human prions in transgenic mice. AB - Transgenic (Tg) mice expressing full-length bovine prion protein (BoPrP) serially propagate bovine spongiform encephalopathy (BSE) prions without posing a transmission barrier. These mice also posed no transmission barrier for Suffolk sheep scrapie prions, suggesting that cattle may be highly susceptible to some sheep scrapie strains. Tg(BoPrP) mice were also found to be susceptible to prions from humans with variant Creutzfeldt-Jakob disease (CJD); on second passage in Tg(BoPrP) mice, the incubation times shortened by 30 to 40 days. In contrast, Tg(BoPrP) mice were not susceptible to sporadic, familial, or iatrogenic CJD prions. While the conformational stabilities of bovine-derived and Tg(BoPrP) passaged BSE prions were similar, the stability of sheep scrapie prions was higher than that found for the BSE prions but lower if the scrapie prions were passaged in Tg(BoPrP) mice. Our findings suggest that BSE prions did not arise from a sheep scrapie strain like the one described here; rather, BSE prions may have arisen spontaneously in a cow or by passage of a scrapie strain that maintains its stability upon passage in cattle. It may be possible to distinguish BSE prions from scrapie strains in sheep by combining conformational stability studies with studies using novel Tg mice expressing a chimeric mouse-BoPrP gene. Single-amino-acid substitutions in chimeric PrP transgenes produced profound changes in incubation times that allowed us to distinguish prions causing BSE from those causing scrapie. PMID- 15827141 TI - Characterization of a recombinant herpes simplex virus 1 designed to enter cells via the IL13Ralpha2 receptor of malignant glioma cells. AB - Malignant glioma tumor cells in situ exhibit on their surfaces the interleukin 13 (IL-13) receptor designated IL13Ralpha2. To target herpes simplex virus 1 to this receptor, we constructed a recombinant virus (R5111) in which the known heparan sulfate binding sites in glycoproteins B and C were deleted and IL-13 was inserted into both glycoproteins C and D. We also transduced a baby hamster kidney cell line lacking the known viral receptors (J1-1) and Vero cells with a plasmid encoding IL13Ralpha2. The J1-1 derivative (J-13R) cell line is susceptible to and replicates the R5111 recombinant virus but not the wild-type parent virus. We report the following. (i) Expression of IL13Ralpha2 was rapidly lost from the surface of transduced cells grown in culture. The loss appeared to be related to ligands present in fetal bovine serum in the medium. None of the malignant glioma cell lines cultivated in vitro and tested to date exhibited the IL13Ralpha2 receptor. (ii) Soluble IL-13 but not IL-4 or IL-2 blocked the replication of R5111 recombinant virus in J-13R cells. (iii) The endocytosis inhibitor PD98059 blocked the replication in J1-1 cells of a mutant lacking glycoprotein D (gD-/-) but not the replication of R5111 in the J-13R cells. We conclude that R5111 enters cells via its interaction with the IL13Ralpha2 receptor in a manner that cannot be differentiated from the interaction of wild type virus with its receptors. PMID- 15827142 TI - Unique long terminal repeat and surface glycoprotein gene sequences of feline leukemia virus as determinants of disease outcome. AB - The outcome of feline leukemia virus (FeLV) infection in nature is variable, including malignant, proliferative, and degenerative disorders. The determinants of disease outcome are not well understood but are thought to include viral, host, and environmental factors. In particular, genetic variations in the FeLV long terminal repeat (LTR) and SU gene have been linked to disease outcome. FeLV 945 was previously identified as a natural isolate predominant in non-T-cell neoplastic and nonneoplastic diseases in a geographic cohort. The FeLV-945 LTR was shown to contain unique repeat elements, including a 21-bp triplication downstream of the enhancer. The FeLV-945 SU gene was shown to encode mutational changes in functional domains of the protein. The present study details the outcomes of infection with recombinant FeLVs in which the LTR and envelope (env) gene of FeLV-945, or the LTR only, was substituted for homologous sequences in a horizontally transmissible prototype isolate, FeLV-A/61E. The results showed that the FeLV-945 LTR determined the kinetics of disease. Substitution of the FeLV-945 LTR into FeLV-A/61E resulted in a significantly more rapid disease onset but did not alter the tumorigenic spectrum. In contrast, substitution of both the FeLV 945 LTR and env gene changed the disease outcome entirely. Further, the impact of FeLV-945 env on the disease outcome was dependent on the route of inoculation. Since the TM genes of FeLV-945 and FeLV-A/61E are nearly identical but the SU genes differ significantly, FeLV-945 SU is implicated in the outcome. These findings identify the FeLV-945 LTR and SU gene as determinants of disease. PMID- 15827143 TI - Identification of novel subgenomic RNAs and noncanonical transcription initiation signals of severe acute respiratory syndrome coronavirus. AB - The expression of the genomic information of severe acute respiratory syndrome coronavirus (SARS CoV) involves synthesis of a nested set of subgenomic RNAs (sgRNAs) by discontinuous transcription. In SARS CoV-infected cells, 10 sgRNAs, including 2 novel ones, were identified, which were predicted to be functional in the expression of 12 open reading frames located in the 3' one-third of the genome. Surprisingly, one new sgRNA could lead to production of a truncated spike protein. Sequence analysis of the leader-body fusion sites of each sgRNA showed that the junction sequences and the corresponding transcription-regulatory sequence (TRS) are unique for each species of sgRNA and are consistent after virus passages. For the two novel sgRNAs, each used a variant of the TRS that has one nucleotide mismatch in the conserved hexanucleotide core (ACGAAC) in the TRS. Coexistence of both plus and minus strands of SARS CoV sgRNAs and evidence for derivation of the sgRNA core sequence from the body core sequence favor the model of discontinuous transcription during minus-strand synthesis. Moreover, one rare species of sgRNA has the junction sequence AAA, indicating that its transcription could result from a noncanonical transcription signal. Taken together, these results provide more insight into the molecular mechanisms of genome expression and subgenomic transcription of SARS CoV. PMID- 15827144 TI - A conformational change in the adeno-associated virus type 2 capsid leads to the exposure of hidden VP1 N termini. AB - The complex infection process of parvoviruses is not well understood so far. An important role has been attributed to a phospholipase A2 domain which is located within the unique N terminus of the capsid protein VP1. Based on the structural difference between adeno-associated virus type 2 wild-type capsids and capsids lacking VP1 or VP2, we show via electron cryomicroscopy that the N termini of VP1 and VP2 are involved in forming globules inside the capsids of empty and full particles. Upon limited heat shock, VP1 and possibly VP2 become exposed on the outsides of full but not empty capsids, which is correlated with the disappearance of the globules in the inner surfaces of the capsids. Using molecular modeling, we discuss the constraints on the release of the globularly organized VP1-unique N termini through the channels at the fivefold symmetry axes outside of the capsid. PMID- 15827145 TI - Complete genome sequence and in planta subcellular localization of maize fine streak virus proteins. AB - The genome of the nucleorhabdovirus maize fine streak virus (MFSV) consists of 13,782 nucleotides of nonsegmented, negative-sense, single-stranded RNA. The antigenomic strand consisted of seven open reading frames (ORFs), and transcripts of all ORFs were detected in infected plants. ORF1, ORF6, and ORF7 had significant similarities to the nucleocapsid protein (N), glycoprotein (G), and polymerase (L) genes of other rhabdoviruses, respectively, whereas the ORF2, ORF3, ORF4, and ORF5 proteins had no significant similarities. The N (ORF1), ORF4, and ORF5 proteins localized to nuclei, consistent with the presence of nuclear localization signals (NLSs) in these proteins. ORF5 likely encodes the matrix protein (M), based on its size, the position of its NLS, and the localization of fluorescent protein fusions to the nucleus. ORF2 probably encodes the phosphoprotein (P) because, like the P protein of Sonchus yellow net virus (SYNV), it was spread throughout the cell when expressed alone but was relocalized to a subnuclear locus when coexpressed with the MFSV N protein. Unexpectedly, coexpression of the MFSV N and P proteins, but not the orthologous proteins of SYNV, resulted in accumulations of both proteins in the nucleolus. The N and P protein relocalization was specific to cognate proteins of each virus. The subcellular localizations of the MFSV ORF3 and ORF4 proteins were distinct from that of the SYNV sc4 protein, suggesting different functions. To our knowledge, this is the first comparative study of the cellular localizations of plant rhabdoviral proteins. This study indicated that plant rhabdoviruses are diverse in genome sequence and viral protein interactions. PMID- 15827146 TI - Array analysis of simian varicella virus gene transcription in productively infected cells in tissue culture. AB - Simian varicella virus (SVV) is a neurotropic alphaherpesvirus of monkeys that is a model for varicella pathogenesis and latency. Like human varicella-zoster virus (VZV), SVV causes chicken pox (varicella), becomes latent in ganglia along the entire neuraxis, and reactivates to produce shingles (zoster). We developed macroarrays to determine the extent of viral transcription from all 70 predicted SVV open reading frames (ORFs) in infected cells in tissue culture. Cloned fragments (200 to 400 bp) from the 5' and 3' ends of each ORF were PCR amplified, quantitated, spotted onto nylon membranes, and fixed by UV cross-linking. Using a cDNA probe prepared from poly(A)+ RNA extracted from SVV-infected Vero cells at the height of the cytopathic effect (3 days after infection) and chemiluminescence for detection, transcripts corresponding to all SVV ORFs were identified. The abundance of each SVV transcript was compared with that previously demonstrated for VZV in infected tissue culture cells. PMID- 15827147 TI - RhoA signaling is required for respiratory syncytial virus-induced syncytium formation and filamentous virion morphology. AB - Respiratory syncytial virus (RSV) is an important human pathogen that can cause severe and life-threatening respiratory infections in infants, the elderly, and immunocompromised adults. RSV infection of HEp-2 cells induces the activation of RhoA, a small GTPase. We therefore asked whether RhoA signaling is important for RSV replication or syncytium formation. The treatment of HEp-2 cells with Clostridium botulinum C3, an enzyme that ADP-ribosylates and specifically inactivates RhoA, inhibited RSV-induced syncytium formation and cell-to-cell fusion, although similar levels of PFU were released into the medium and viral protein expression levels were equivalent. Treatment with another inhibitor of RhoA signaling, the Rho kinase inhibitor Y-27632, yielded similar results. Scanning electron microscopy of C3-treated infected cells showed reduced numbers of single blunted filaments, in contrast to the large clumps of long filaments in untreated infected cells. These data suggest that RhoA signaling is associated with filamentous virus morphology, cell-to-cell fusion, and syncytium formation but is dispensable for the efficient infection and production of infectious virus in vitro. Next, we developed a semiquantitative method to measure spherical and filamentous virus particles by using sucrose gradient velocity sedimentation. Fluorescence and transmission electron microscopy confirmed the separation of spherical and filamentous forms of infectious virus into two identifiable peaks. The C3 treatment of RSV-infected cells resulted in a shift to relatively more spherical virions than those from untreated cells. These data suggest that viral filamentous protuberances characteristic of RSV infection are associated with RhoA signaling, are important for filamentous virion morphology, and may play a role in initiating cell-to-cell fusion. PMID- 15827148 TI - Structure and assembly of a T=1 virus-like particle in BK polyomavirus. AB - In polyomaviruses the pentameric capsomers are interlinked by the long C-terminal arm of the structural protein VP1. The T=7 icosahedral structure of these viruses is possible due to an intriguing adaptability of this linker arm to the different local environments in the capsid. To explore the assembly process, we have compared the structure of two virus-like particles (VLPs) formed, as we found, in a calcium-dependent manner by the VP1 protein of human polyomavirus BK. The structures were determined using electron cryomicroscopy (cryo-EM), and the three dimensional reconstructions were interpreted by atomic modeling. In the small VP1 particle, 26.4 nm in diameter, the pentameric capsomers form an icosahedral T=1 surface lattice with meeting densities at the threefold axes that interlinked three capsomers. In the larger particle, 50.6 nm in diameter, the capsomers form a T=7 icosahedral shell with three unique contacts. A folding model of the BKV VP1 protein was obtained by alignment with the VP1 protein of simian virus 40 (SV40). The model fitted well into the cryo-EM density of the T=7 particle. However, residues 297 to 362 of the C-terminal arm had to be remodeled to accommodate the higher curvature of the T=1 particle. The loops, before and after the C-terminal short helix, were shown to provide the hinges that allowed curvature variation in the particle shell. The meeting densities seen at the threefold axes in the T=1 particle were consistent with the triple-helix interlinking contact at the local threefold axes in the T=7 structure. PMID- 15827149 TI - St, a truncated envelope protein derived from the S protein of duck hepatitis B virus, acts as a chaperone for the folding of the large envelope protein. AB - Envelope proteins of hepadnaviruses undergo a unique folding mechanism which results in the posttranslational translocation of 50% of the large envelope protein (L) chains across the endoplasmic reticulum. This mechanism is essential for the eventual positioning of the receptor-binding domain on the surface of the virus particle and in duck hepatitis B virus (DHBV) is dependent on the small (S) envelope protein as part of the assembly process. In this study, we report the identification of a third envelope protein, St, derived from the S protein and carrying functions previously attributed to S. Antibody mapping and mutagenesis studies indicated St to be C terminally truncated, spanning the N-terminal transmembrane domain (TM1) plus the adjacent cysteine loop. We have previously shown that the mutation of two conserved polar residues in TM1 of S (SAA) eliminates L translocation and assembly. A plasmid expressing a functional equivalent of St was able to rescue assembly, demonstrating that this assembly defect is due to mutations of the corresponding residues in St and not in S per se. Immunofluorescence analysis showed that St directly affects L protein cellular localization. These results indicate that St acts as a viral chaperone for L folding, remaining associated with the DHBV envelope upon secretion. The presence of St at a molar ratio of half that of L suggests that it is St which regulates L translocation to 50%. PMID- 15827150 TI - Effects of nonstructural proteins NS1 and NS2 of human respiratory syncytial virus on interferon regulatory factor 3, NF-kappaB, and proinflammatory cytokines. AB - Human respiratory syncytial virus (HRSV) is the leading cause of serious pediatric acute respiratory tract infections, and a better understanding is needed of the host response to HRSV and its attenuated vaccine derivatives. It has been shown previously that HRSV nonstructural proteins 1 and 2 (NS1 and NS2) inhibit the induction of alpha/beta interferon (IFN-alpha/beta) in A549 cells and human macrophages. Two principal transcription factors for the early IFN-beta and -alpha1 response are interferon regulatory factor 3 (IRF-3) and nuclear factor kappaB (NF-kappaB). At early times postinfection, wild-type HRSV and the NS1/NS2 deletion mutants were very similar in the ability to activate IRF-3. However, once NS1 and NS2 were expressed significantly, they acted cooperatively to suppress activation and nuclear translocation of IRF-3. Since these viruses differed greatly in the induction of IFN-alpha/beta, NF-kappaB activation was evaluated in Vero cells, which lack the structural genes for IFN-alpha/beta and would preclude confounding effects of IFN-alpha/beta. This showed that deletion of the NS2 gene sharply reduced the ability of HRSV to induce activation of NF kappaB. Since recombinant HRSVs from which the NS1 or NS2 genes have been deleted are being developed as vaccine candidates, we investigated whether the changes in activation of host transcription factors and increased IFN-alpha/beta production had an effect on the epithelial production of proinflammatory factors. Viruses lacking NS1 and/or NS2 stimulated modestly lower production of RANTES (Regulated on Activation Normal T-cell Expressed and Secreted), interleukin 8, and tumor necrosis factor alpha compared to wild-type recombinant RSV, supporting their use as attenuated vaccine candidates. PMID- 15827151 TI - Amino acid changes in proteins 2B and 3A mediate rhinovirus type 39 growth in mouse cells. AB - Many steps of viral replication are dependent on the interaction of viral proteins with host cell components. To identify rhinovirus proteins involved in such interactions, human rhinovirus 39 (HRV39), a virus unable to replicate in mouse cells, was adapted to efficient growth in mouse cells producing the viral receptor ICAM-1 (ICAM-L cells). Amino acid changes were identified in the 2B and 3A proteins of the adapted virus, RV39/L. Changes in 2B were sufficient to permit viral growth in mouse cells; however, changes in both 2B and 3A were required for maximal viral RNA synthesis in mouse cells. Examination of infected HeLa cells by electron microscopy demonstrated that human rhinoviruses induced the formation of cytoplasmic membranous vesicles, similar to those observed in cells infected with other picornaviruses. Vesicles were also observed in the cytoplasm of HRV39 infected mouse cells despite the absence of viral RNA replication. Synthesis of picornaviral nonstructural proteins 2C, 2BC, and 3A is known to be required for formation of membranous vesicles. We suggest that productive HRV39 infection is blocked in ICAM-L cells at a step posttranslation and prior to the formation of a functional replication complex. The observation that changes in HRV39 2B and 3A proteins lead to viral growth in mouse cells suggests that one or both of these proteins interact with host cell proteins to promote viral replication. PMID- 15827152 TI - Gamma interferon-dependent, noncytolytic clearance of sindbis virus infection from neurons in vitro. AB - Due to the nonrenewable nature of neurons, recovery from viral infection of the central nervous system requires noncytopathic mechanisms for control of virus replication. Recovery from alphavirus encephalitis can occur without apparent neurological damage through the effects of antibody and gamma interferon (IFN gamma). To establish an in vitro cell culture system that will allow the study of mechanisms of IFN-gamma-mediated control of Sindbis virus (SINV) replication in neurons, we have characterized the susceptibility to SINV infection and IFN-gamma responsiveness of two neuronal cell lines that can be differentiated in vitro: CSM14.1, a rat nigral cell line, and NSC34, a mouse motor neuron cell line. Undifferentiated CSM14.1 and NSC34 cells were permissive for SINV and susceptible to virus-induced cell death. With differentiation, CSM14.1 cells reduced virus replication and became progressively resistant to virus-induced cell death, resulting in prolonged virus replication. NSC34 cells did not differentiate completely and became only partially resistant to SINV infection. Both CSM14.1 and NSC34 cells responded to pretreatment with IFN-gamma by decreasing SINV replication. Differentiated CSM14.1 cells treated 24 h after infection with IFN gamma responded with increased cell viability and clearance of infectious virus. IFN-gamma treatment sequentially altered the ratio of genomic to subgenomic viral RNA synthesis, promoted recovery of cellular protein synthesis, reduced viral protein synthesis, and inhibited viral RNA transcription within 24 h after treatment. We conclude that CSM14.1 cells provide an excellent model for the study of IFN-gamma-mediated noncytolytic clearance of SINV from mature neurons. PMID- 15827153 TI - Covert human immunodeficiency virus replication in dendritic cells and in DC-SIGN expressing cells promotes long-term transmission to lymphocytes. AB - HIV-1 virions are efficiently captured by monocyte-derived immature dendritic cells (iDCs), as well as by cell lines expressing the lectin DC-SIGN. Viral infectivity can be retained for several days, and even enhanced, before transmission to CD4+ lymphocytes. The role of DC-SIGN in viral retention and enhancement of infection is not fully understood and varies according to the cell line expressing the lectin. We studied here the mechanisms underlying this process. We focused our study on X4-tropic human immunodeficiency virus (HIV) strains, since they were widely believed not to replicate in iDCs. However, we first show that X4 HIV replicates covertly and slowly in iDCs. This is also the case in Raji-DC-SIGN cells, which are classically used to study HIV transmission. We used either single-cycle or replicative HIV and measured viral RT and replication to further demonstrate that transfer of incoming virions from iDCs or DC-SIGN+ cells occurs only on the short-term (i.e., a few hours after viral exposure). There is no long-term storage of original HIV particles in these cells. A few days after viral exposure, replicative viruses, and not single-cycle virions, are transmitted to CD4+ cells. The cell-type-dependent activity of DC SIGN reflects the ability of HIV to replicate covertly in some cells, and not in others. PMID- 15827154 TI - Highly protective in vivo function of cytomegalovirus IE1 epitope-specific memory CD8 T cells purified by T-cell receptor-based cell sorting. AB - Reconstitution of antiviral CD8 T cells is essential for controlling cytomegalovirus (CMV) infection after bone marrow transplantation. Accordingly, polyclonal CD8 T cells derived from BALB/c mice infected with murine CMV protect immunocompromised adoptive transfer recipients against CMV disease. The protective population comprises CD8 T cells with T-cell receptors (TCRs) specific for defined and for as-yet-unknown viral epitopes, as well as a majority of nonprotective cells with unrelated specificities. Defined epitopes include IE1/m123 and m164, which are immunodominant in terms of the magnitude of the CD8 T-cell response, and a panel of subordinate epitopes (m04, m18, M45, M83, and M84). While cytolytic T-lymphocyte lines (CTLLs) were shown to be protective regardless of the immunodominance of the respective epitope, the individual contributions of in vivo resident epitope-specific CD8 T cells to the antiviral control awaited investigation. The IE1 peptide 168-YPHFMPTNL-176 is generated from the immediate-early protein 1 (IE1) (pp89/76) of murine CMV and is presented by the major histocompatibility complex class I (MHC-I) molecule Ld. To quantitate its contribution to the protective potential of a CD8-T memory (CD8 TM) cell population, IE1-TCR+ and IE1-TCR- CD8-TM cells were purified by epitope specific cell sorting with IE1 peptide-loaded MHC-immunoglobulin G1 dimers as ligands of cognate TCRs. Of relevance for clinical approaches to an adoptive cellular immunotherapy, sorted IE1 epitope-specific CD8-TM cells were found to be exceedingly protective upon adoptive transfer. Compared with CTLLs specific for the same epitope and of comparable avidity and TCR beta-chain variable region (Vbeta)-defined polyclonality, sorted CD8-TM cells proved to be superior by more than 2 orders of magnitude. PMID- 15827155 TI - Dengue virus inhibits alpha interferon signaling by reducing STAT2 expression. AB - Alpha/beta interferon (IFN-alpha/beta) is a key mediator of innate antiviral responses but has little effect on the established replication of dengue viruses, which are mosquito-borne flaviviruses of immense global health importance. Understanding how the IFN system is inhibited in dengue virus-infected cells would provide critical insights into disease pathogenesis. In a recent study analyzing the ability of individual dengue virus-encoded proteins to antagonize the IFN response, nonstructural (NS) protein 4B and possibly NS2A and NS4A were identified as candidate IFN antagonists. In monkey cells, NS4B appeared to inhibit both the IFN-alpha/beta and IFN-gamma signal transduction pathways, which are distinct but overlapping (J. L. Munoz-Jordan, G. G. Sanchez-Burgos, M. Laurent-Rolle, and A. Garcia-Sastre, Proc. Natl. Acad. Sci. USA 100:14333-14338, 2003). For this study, we examined the effects of dengue virus on the human IFN system, using cell lines that were stably transfected with self-replicating subgenomic dengue virus RNA (replicons) and that expressed all of the dengue virus nonstructural proteins together. We show here that in replicon-containing cells dengue virus RNA replication and the replication of encephalomyocarditis virus, an IFN-sensitive virus, are resistant to the antiviral effects of IFN alpha. The presence of dengue virus replicons reduces global IFN-alpha-stimulated gene expression and specifically inhibits IFN-alpha but not IFN-gamma signal transduction. In cells containing replicons or infected with dengue virus, we found reduced levels of signal transducer and activator of transcription 2 (STAT2), which is a key component of IFN-alpha but not IFN-gamma signaling. Collectively, these data show that dengue virus is capable of subverting the human IFN response by down-regulating STAT2 expression. PMID- 15827156 TI - Few mutations in the 5' leader region mediate fitness recovery of debilitated human immunodeficiency type 1 viruses. AB - Repeated bottleneck passages of RNA viruses result in fitness losses due to the accumulation of deleterious mutations. In contrast, repeated transfers of large virus populations result in exponential fitness increases. Human immunodeficiency virus type 1 (HIV-1) manifested a drastic fitness loss after a limited number of plaque-to-plaque transfers in MT-4 cells. An analysis of the mutations associated with fitness loss in four debilitated clones revealed mutation frequencies in gag that were threefold higher than those in env. We now show an increase in the fitness of the debilitated HIV-1 clones by repeated passages of large populations. An analysis of the entire genomic nucleotide sequences of these populations showed that few mutations, from two to seven per clone, mediated fitness recovery. Eight of the 20 mutations affected coding regions, mainly by the introduction of nonsynonymous mutations (75%). However, most of the mutations accumulated during fitness recovery (12 of 20) were located in the 5' untranslated leader region of the genome, and more specifically, in the primer binding site (PBS) loop. Two of the viruses incorporated the same mutation in the primer activation signal in the PBS loop, which is critical for the tRNA3Lys mediated initiation of reverse transcription. Moreover, 25% of the mutations observed were reversions. This fact, together with the presence of a large proportion of nonsynonymous replacements, may disclose the operation, during large population passages, of strong positive selection for optimal HIV-1 replication, which seems to be primarily affected by binding of the tRNA to the PBS and the initiation of reverse transcription. PMID- 15827157 TI - Viremia and nasal and rectal shedding of rotavirus in gnotobiotic pigs inoculated with Wa human rotavirus. AB - Respiratory symptoms with rotavirus shedding in nasopharyngeal secretions have been reported in children with and without gastrointestinal symptoms (Zheng et al., 1991, J. Med. Virol. 34:29-37). To investigate if attenuated and virulent human rotavirus (HRV) strains cause upper respiratory tract infections or viremia in gnotobiotic pigs, we inoculated them with attenuated or virulent HRV intranasally, intravenously, or orally or via feeding tube (gavage) and assayed virus shedding. After oral or intranasal inoculation with attenuated HRV, the pigs remained asymptomatic, but 79 to 95% shed virus nasally and 5 to 17% shed virus rectally. After inoculation by gavage, no pigs shed virus nasally or rectally, but all pigs seroconverted with antibodies to HRV. No viremia was detected through postinoculation day 10. Controls inoculated intranasally with nonreplicating rotavirus-like particles or mock inoculated did not shed virus. In contrast, 100% of pigs inoculated with virulent HRV (oral, intranasal, or gavage) developed diarrhea, shed virus nasally and rectally, and had viremia. The infectivity of sera from the viremic virulent HRV-inoculated pigs was confirmed by inoculating gnotobiotic pigs orally with pooled HRV-positive serum. Serum inoculated pigs developed diarrhea and fecal and nasal virus shedding and seroconverted with serum and intestinal HRV antibodies. Pigs inoculated intravenously with serum or intestinal contents from the viremic virulent HRV inoculated pigs developed diarrhea, virus shedding, and viremia, similar to the orally inoculated pigs. This study provides new evidence that virulent HRV causes transient viremia and upper respiratory tract infection in addition to gastrointestinal infection in gnotobiotic pigs, confirming previous reports of rotavirus antigenemia (Blutt et al., Lancet 362:1445-1449, 2003). Our data also suggest that intestinal infection might be initiated from the basolateral side of the epithelial cells via viremia. Additionally, virus shedding patterns indicate a different pathogenesis for attenuated versus virulent HRV. PMID- 15827158 TI - Replication-competent adenovirus formation in 293 cells: the recombination-based rate is influenced by structure and location of the transgene cassette and not increased by overproduction of HsRad51, Rad51-interacting, or E2F family proteins. AB - Propagation of E1 region replacement adenovirus vectors in 293 cells results in the rare appearance of replication-competent adenovirus (RCA). The RCA genome contains E1 DNA acquired from the 293 cellular genome. The Luria-Delbruck fluctuation test was adapted to measure RCA formation rates. To test if structure affected rate, we measured rates during the production of adenovirus vectors with genomes containing three different expression cassette arrangements. The vectors had different extents of sequence identity with integrated Ad5 DNA of 293 cells and had different distributions of identity flanking the expression cassettes. Empty cassette vector RCA rates ranged from 2.5 x 10(-8) to 5.6 x 10(-10). The extent of sequence identity was not an accurate RCA rate predictor. The vector with the highest RCA rate also had the least overall sequence identity. To define factors controlling RCA generation, adenovirus vectors expressing E2F family proteins, known to modulate recombination gene expression, and overexpressing the human Rad51 recombination protein were analyzed. Compared to their corresponding empty vectors, RCA rates were not increased but were slightly decreased. Initial results suggested expression cassette orientation and/or transcription direction as potential RCA rate modifiers. Testing adenovirus vectors with identical transgene cassettes oriented in opposite directions suggested that transcription direction was not the basis of these rate differences. Thus, the overall structure and location of the transgene cassette had the largest effect on RCA rate. The RCA fluctuation test should be useful for investigators who require accurate measurements of targeted recombination and the probability of RCA formation during stock production. PMID- 15827159 TI - Potential of equine herpesvirus 1 as a vector for immunization. AB - Key problems using viral vectors for vaccination and gene therapy are antivector immunity, low transduction efficiencies, acute toxicity, and limited capacity to package foreign genetic information. It could be demonstrated that animal and human cells were efficiently transduced with equine herpesvirus 1 (EHV-1) reconstituted from viral DNA maintained and manipulated in Escherichia coli. Between 13 and 23% of primary human CD3+, CD4+, CD8+, CD11b+, and CD19+ cells and more than 70% of CD4+ MT4 cells or various human tumor cell lines (MeWo, Huh7, HeLa, 293T, or H1299) could be transduced with one infectious unit of EHV-1 per cell. After intranasal instillation of EHV-1 into mice, efficient transgene expression in lungs was detectable. Successful immunization using EHV-1 was shown after delivery of the human immunodeficiency virus type 1 Pr55gag precursor by the induction of a Gag-specific CD8+ immune response in mice. Because EHV-1 was not neutralized by human sera containing high titers of antibodies directed against human herpesviruses 1 to 5, it is concluded that this animal herpesvirus has enormous potential as a vaccine vector, because it is able to efficiently transduce a variety of animal and human cells, has high DNA packaging capacity, and can conveniently be maintained and manipulated in prokaryotic cells. PMID- 15827160 TI - Linked tumor-selective virus replication and transgene expression from E3 containing oncolytic adenoviruses. AB - Historically, the adenoviral E3 region was found to be nonessential for viral replication in vitro. In addition, adenoviruses whose genome was more than approximately 105% the size of the native genome were inefficiently packaged. These profound observations were used experimentally to insert transgenes into the adenoviral backbone. More recently, however, the reintroduction of the E3 region into oncolytic adenoviruses has been found to positively influence antitumor efficacy in preclinical models and clinical trials. In the studies reported here, the granulocyte-macrophage colony-stimulating factor (GM-CSF) cDNA sequence has been substituted for the E3-gp19 gene in oncolytic adenoviruses that otherwise retained the E3 region. Five viruses that differed slightly in the method of transgene insertion were generated and compared to Ar6pAE2fGmF (E2F/GM/DeltaE3), a previously described E3-deleted oncolytic adenovirus encoding GM-CSF. In all of the viruses, the human E2F-1 promoter regulated E1A expression and GM-CSF expression was under the control of the adenoviral E3 promoter and the packaging signal was relocated immediately upstream from the right terminal repeat. The E3-gp19-deleted viruses had similar cytolytic properties, as measured in vitro by cytotoxicity assays, but differed markedly in their capacity to express and secrete GM-CSF. Ar15pAE2fGmF (E2F/GM/E3b), the virus that produced the highest levels of GM-CSF and retained the native GM-CSF leader sequence, was selected for further analysis. The E2F/GM/E3b and E2F/GM/DeltaE3 viruses exhibited similar cytotoxic activity and GM-CSF production in several tumor cell lines in vitro. However, when compared in vivo in nude mouse xenograft tumor models, E2F/GM/E3b spread through tumors to a greater extent, resulted in higher peak GM-CSF and total exposure levels in both tumor and serum, and was more efficacious than the E3-deleted virus. Using the matched WI-38 (parental) and WI 38-VA13 (simian virus 40 large T antigen transformed) cell pair, GM-CSF was shown to be selectively produced in cells expressing high levels of E2F, indicating that the tumor-selective E2F promoter controlled E1A and GM-CSF expression. PMID- 15827161 TI - Characterization of prototype foamy virus gag late assembly domain motifs and their role in particle egress and infectivity. AB - Foamy viruses (FV) are unusual among retroviruses since they require both Gag and Env structural proteins for particle egress. Recently significant progress has been made towards the mechanistic understanding of the viral release process, in particular that of retroviruses, and the viral domains and cellular pathways involved. However little is currently known about domains of FV structural proteins and cellular proteins engaged in this process. By mutational analysis of sequence motifs in prototype FV (PFV) Gag, bearing homology to known late assembly (L) domains, a PSAP motif with L domain function that was functionally interchangeable by heterologous L domains was identified. In contrast the inactivation of a PPPI motif had no significant influence on PFV particle release, although mutant viral particles displayed reduced infectivity. Similarly mutation of an evolutionary conserved YXXL motif revealed no classical L-domain function but resulted in release of noninfectious viruslike particles. Biochemical and electron microscopy analysis demonstrated that these mutant particles incorporated all viral structural proteins but contained aberrantly capsid structures, suggesting a role in capsid assembly for this PFV Gag sequence motif. In line with the mutational analysis, overexpression of dominant negative (DN) mutants and wild-type TSG101 but not the DN mutant of AIP-1/ALIX reduced PFV particle release and infectivity. Furthermore, DN mutants of Vps4A, Vps4B, and CHMP3 inhibited PFV egress and infectivity. Taken together these results demonstrate that PFV, like other viruses, requires components of the vacuolar protein sorting (VPS) machinery for egress and enters the VPS pathway through interaction with TSG101. PMID- 15827162 TI - Regression of Epstein-Barr virus-induced B-cell transformation in vitro involves virus-specific CD8+ T cells as the principal effectors and a novel CD4+ T-cell reactivity. AB - T-cell memory to Epstein-Barr virus (EBV) was first demonstrated through regression of EBV-induced B-cell transformation to lymphoblastoid cell lines (LCLs) in virus-infected peripheral blood mononuclear cell (PBMC) cultures. Here, using donors with virus-specific T-cell memory to well-defined CD4 and CD8 epitopes, we reexamine recent reports that the effector cells mediating regression are EBV latent antigen-specific CD4+ and not, as previously assumed, CD8+ T cells. In regressing cultures, we find that the reversal of CD23+ B-cell proliferation was always coincident with an expansion of latent epitope-specific CD8+, but not CD4+, T cells; furthermore CD8+ T-cell clones derived from regressing cultures were epitope specific and reproduced regression when cocultivated with EBV-infected autologous B cells. In cultures of CD4-depleted PBMCs, there was less efficient expansion of these epitope-specific CD8+ T cells and correspondingly weaker regression. The data are consistent with an effector role for epitope-specific CD8+ T cells in regression and an auxiliary role for CD4+ T cells in expanding the CD8 response. However, we also occasionally observed late regression in CD8-depleted PBMC cultures, though again without any detectable expansion of preexisting epitope-specific CD4+ T-cell memory. CD4+ T cell clones derived from such cultures were LCL specific in gamma interferon release assays but did not recognize any known EBV latent cycle protein or derived peptide. A subset of these clones was also cytolytic and could block LCL outgrowth. These novel effectors, whose antigen specificity remains to be determined, may also play a role in limiting virus-induced B-cell proliferation in vitro and in vivo. PMID- 15827163 TI - Human immunodeficiency virus type 1 inhibits DNA damage-triggered apoptosis by a Nef-independent mechanism. AB - It is controversial whether the accessory human immunodeficiency virus type 1 (HIV-1) Nef protein inhibits or enhances apoptosis. To address this issue, we investigated the effect of Nef on programmed cell death with vectors or proviral HIV-1 constructs coexpressing Nef and green fluorescent protein from single bicistronic RNAs. This approach allows us to readily identify transfected or infected cells and to correlate cell death directly with Nef expression levels. We demonstrate that Nef does not significantly affect apoptosis in transfected or HIV-1-infected Jurkat T cells or primary human peripheral blood mononuclear cells. Unexpectedly, however, both nef+ and nef-defective HIV-1 infection blocked apoptosis in cells treated with UV light or etoposide but not cell death induced by CD95 antibody, TRAIL, Ly294002, or serum starvation. Our results show that HIV 1 infection inhibits DNA damage-induced but not death receptor-dependent cell death by a Nef-independent mechanism. PMID- 15827164 TI - Modulation of the cell growth regulator mTOR by Epstein-Barr virus-encoded LMP2A. AB - Control of translation initiation is one means by which cells regulate growth and proliferation, with components of the protein-synthesizing machinery having oncogenic potential. Expression of latency protein LMP2A by the human tumor virus Epstein-Barr virus (EBV) activates phosphatidylinositol 3-kinase/Akt located upstream of an essential mediator of growth signals, mTOR (mammalian target of rapamycin). We show that mTOR is activated by expression of LMP2A in carcinoma cells, leading to wortmannin- and rapamycin-sensitive inhibition of the negative regulator of translation, eukaryotic initiation factor 4E-binding protein 1, and increased c-Myc protein translation. Intervention by this DNA tumor virus in cellular translational controls is likely to be an integral component of EBV tumorigenesis. PMID- 15827165 TI - Inhibition of toll-like receptor 7- and 9-mediated alpha/beta interferon production in human plasmacytoid dendritic cells by respiratory syncytial virus and measles virus. AB - Human plasmacytoid dendritic cells (PDC) are key sentinels alerting both innate and adaptive immune responses through production of huge amounts of alpha/beta interferon (IFN). IFN induction in PDC is triggered by outside-in signal transduction pathways through Toll-like receptor 7 (TLR7) and TLR9 as well as by recognition of cytosolic virus-specific patterns. TLR7 and TLR9 ligands include single-stranded RNA and CpG-rich DNA, respectively, as well as synthetic derivatives thereof which are being evaluated as therapeutic immune modulators promoting Th1 immune responses. Here, we identify the first viruses able to block IFN production by PDC. Both TLR-dependent and -independent IFN responses are abolished in human PDC infected with clinical isolates of respiratory syncytial virus (RSV), RSV strain A2, and measles virus Schwarz, in contrast to RSV strain Long, which we previously identified as a potent IFN inducer in human PDC (Hornung et al., J. Immunol. 173:5935-5943, 2004). Notably, IFN synthesis of PDC activated by the TLR7 and TLR9 agonists resiquimod (R848) and CpG oligodeoxynucleotide 2216 is switched off by subsequent infection by RSV A2 and measles virus. The capacity of RSV and measles virus of human PDC to shut down IFN production should contribute to the characteristic features of these viruses, such as Th2-biased immune pathology, immune suppression, and superinfection. PMID- 15827166 TI - Recombinant, live-attenuated tetravalent dengue virus vaccine formulations induce a balanced, broad, and protective neutralizing antibody response against each of the four serotypes in rhesus monkeys. AB - Three tetravalent vaccine (TV) formulations of previously described monovalent dengue (DEN) virus vaccine candidates were compared to a tetravalent formulation of wild-type DEN viruses (T-wt) for replication in SCID mice transplanted with human liver cells (SCID-HuH-7) or for replication and immunogenicity in rhesus monkeys. TV-1 consists of recombinant DEN1, -2, -3, and -4, each with a 30 nucleotide deletion in the 3' untranslated region (Delta30). TV-2 consists of rDEN1Delta30, rDEN4Delta30, and two antigenic chimeric viruses, rDEN2/4Delta30 and rDEN3/4Delta30, both also bearing the Delta30 mutation. TV-3 consists of rDEN1Delta30, rDEN2Delta30, rDEN4Delta30, and a 10-fold higher dose of rDEN3/4Delta30. TV-1 and TV-2 were attenuated in SCID-HuH-7 mice with minimal interference in replication among the virus components. TV-1, -2, and -3 were attenuated in rhesus monkeys as measured by duration and peak of viremia. Each monkey immunized with TV-1 and TV-3 seroconverted to the four DEN components by day 28 with neutralization titers ranging from 1:52 to 1:273 and 1:59 to 1:144 for TV-1 and TV-3, respectively. TV-2 induced low antibody titers to DEN2 and DEN3, but a booster immunization after 4 months increased the neutralizing antibody titers to greater than 1:100 against each serotype and elicited broad neutralizing activity against 19 of 20 DEN subtypes. A single dose of TV-2 induced protection against wild-type DEN1, DEN3, and DEN4 challenge, but not DEN2. However, two doses of TV-2 or TV-3 induced protection against DEN2 challenge. Two tetravalent formulations, TV-2 and TV-3, possess properties of a successful DEN vaccine and can be considered for evaluation in clinical trials. PMID- 15827167 TI - Cross-reactive cytotoxic T lymphocytes against human immunodeficiency virus type 1 protease and gamma interferon-inducible protein 30. AB - The gamma interferon (IFN-gamma)-inducible protein 30 (IP-30) signal peptide -11 to -3 (LLDVPTAAV) is a prominent self peptide expressed with the class I human histocompatibility leukocyte antigen A2 (HLA-A2). Stimulation of peripheral blood mononuclear cells (PBMC) from HLA-A2 human immunodeficiency virus type 1 (HIV-1) infected individuals with an HLA-A2-restricted HIV protease (PR) peptide 76-84 (LVGPTPVNI) activated cytotoxic T lymphocytes (CTL) against the IP-30 signal peptide. Since HIV-1 PR 76-84 stimulated CD8+ T cells from these individuals to secrete IFN-gamma, we tested whether the activation of IP-30-specific CTL in vitro resulted from T-cell cross-reactivity or from up-regulation of IP-30 by IFN gamma. Neither high levels of exogenous IFN-gamma nor incubation of PBMC with other HIV peptides triggering substantial IFN-gamma release activated IP-30 specific CTL. Although the IP-30 signal peptide did not stimulate IFN-gamma release from freshly isolated PBMC, it activated CTL in vitro against itself and HIV PR 76-84. Peptide-stimulated IFN-gamma release, cold target inhibition, and HLA-A2/immunoglobulin dimer-mediated binding and depletion of effector cells all indicated that in vitro stimulation with HIV PR 76-84 or the IP-30 signal peptide activated a comparable population of cross-reactive effector cells. Neither IP-30 nor HIV PR 76-84 activated CTL against themselves following in vitro stimulation of PBMC from non-HIV-infected HLA-A2 individuals. Peptide titrations indicated higher-avidity T-cell interactions with HIV PR 76-84 than with the IP-30 signal peptide. These data indicate that HIV PR 76-84 is a heteroclitic variant of the IP-30 signal peptide -11 to -3, which has implications for immune memory and autoimmunity. PMID- 15827168 TI - Pseudovirion particle production by live poxvirus human immunodeficiency virus vaccine vector enhances humoral and cellular immune responses. AB - Live-vector-based human immunodeficiency virus (HIV) vaccines are an integral part of a number of HIV vaccine regimens currently under evaluation. Live vectors that carry an intact gag gene are capable of eliciting HIV pseudovirion particle formation from infected host cells. The impact of pseudovirion particle formation on the immune response generated by live HIV vaccine vectors has not been established. In this study, a canarypox HIV vaccine candidate vector expressing HIV gag and env genes, vCP205, was modified by the introduction of a glycine-to alanine coding change in the N-terminal myristylation site of gag to create Myr- vCP205. This substitution effectively eliminated particle formation without altering the level of protein production. vCP205 and Myr- vCP205 were then directly compared for the ability to induce HIV-specific immune responses in mice. The particle-competent vector vCP205 elicited higher levels of CD8+ T-cell responses, as indicated by gamma interferon enzyme-linked immunospot (ELISPOT) assay and intracellular cytokine staining. Humoral responses to Gag and Env were also markedly higher from animals immunized with the particle-competent vector. Furthermore, HIV-specific CD4+ T-cell responses were greater among animals immunized with the particle-competent vector. Using a human dendritic cell model of antigen presentation in vitro, vCP205 generated greater ELISPOT responses than Myr- vCP205. These results demonstrate that pseudovirion particle production by live-vector HIV vaccines enhances HIV-specific cellular and humoral immune responses. PMID- 15827169 TI - The transcriptional transactivation function of HBx protein is important for its augmentation role in hepatitis B virus replication. AB - The role and functional domain of hepatitis B virus (HBV) X protein (HBx) in regulating HBV transcription and replication were investigated with a transient transfection system in the human hepatoma cell line HepG2 using wild-type or HBx minus HBV genome constructs and a series of deletion or mutation HBx expression plasmids. We show here that HBx has augmentation effects on HBV transcription and replication as a HBV mutant genome with defective X gene led to decreased levels of 3.5-kb HBV RNA and HBV replication intermediates and that these decreases can be restored by either transient ectopic expression of HBx or a stable HBx expression cell line. The C-terminal two-thirds (amino acids [aa] 51 to 154), which contain the transactivation domain, is required for this function of HBx; the N-terminal one-third (aa 1 to 50) is not required. Using the alanine scanning mutagenesis strategy, we demonstrated that the regions between aa 52 to 65 and 88 to 154 are important for the augmentation function of HBx in HBV replication. By the luciferase reporter gene analysis, we found that the transactivation and coactivation activities of HBx coincide well with its augmentation function in HBV transcription and replication. These results suggest that HBx has an important role in stimulating HBV transcription and replication and that the transcriptional transactivation function of HBx may be critical for its augmentation effect on HBV replication. PMID- 15827170 TI - ATPgammaS disrupts human immunodeficiency virus type 1 virion core integrity. AB - Heat shock protein 70 (Hsp70) is incorporated within the membrane of primate lentiviral virions. Here we demonstrate that Hsp70 is also incorporated into oncoretroviral virions and that it remains associated with membrane-stripped human immunodeficiency virus type 1 (HIV-1) virion cores. To determine if Hsp70 promotes virion infectivity, we attempted to generate Hsp70-deficient virions with gag deletion mutations, Hsp70 transdominant mutants, or RNA interference, but these efforts were confounded, largely because they disrupt virion assembly. Given that polypeptide substrates are bound and released by Hsp70 in an ATP hydrolytic reaction cycle, we supposed that incubation of HIV-1 virions with ATP would perturb Hsp70 interaction with substrates in the virion and thereby decrease infectivity. Treatment with ATP or ADP had no observable effect, but ATPgammaS and GTPgammaS, nucleotide triphosphate analogues resistant to Hsp70 hydrolysis, dramatically reduced the infectivity of HIV-1 and murine leukemia virus virions. ATPgammaS-treated virions were competent for fusion with susceptible target cells, but viral cDNA synthesis was inhibited to an extent that correlated with the magnitude of decrease in infectivity. Intravirion reverse transcription by HIV-1, simian immunodeficiency virus, or murine leukemia virus was also inhibited by ATPgammaS. The effects of ATPgammaS on HIV-1 reverse transcription appeared to be indirect, resulting from disruption of virion core morphology that was evident by transmission electron microscopy. Consistent with effects on capsid conformation, ATPgammaS-treated viruslike particles failed to saturate host antiviral restriction activity. Our observations support a model in which the catalytic activity of virion-associated Hsp70 is required to maintain structural integrity of the virion core. PMID- 15827172 TI - Novel macula-like virus identified in Bombyx mori cultured cells. AB - We identified a novel, 6,513-bp-long RNA, termed Bombyx mori macula-like latent virus (BmMLV) RNA, which abundantly expressed in B. mori cultured BmN cells. BmMLV RNA potentially encodes two proteins, putative RNA replicase and coat protein, which share structural features and sequence similarities with those of a plant RNA virus, the genus Maculavirus. Northern blot analysis showed that two transcripts were expressed in BmN cells: a 6.5-kb-long RNA, which contains both putative RNA replicase and coat protein genes, and a 1.2-kb-long RNA, which contains only a coat protein gene. Southern blot analysis showed that BmMLV RNA is not carried by the B. mori genome. RT-PCR analysis also revealed the presence of BmMLV RNA in several B. mori cell lines other than BmN cells, suggesting that BmMLV RNA latently exists in B. mori cultured cells. Infection studies showed that BmMLV virions were able to infect BmMLV-negative Spodoptera frugiperda Sf-9 cells and B. mori larvae. Electron microscopy and Northern blot analysis of a purified BmMLV revealed that isometric virions appear to be 28 to 30 nm in diameter and contain a 6.5-kb genomic RNA. These results showed that BmMLV is a novel macula-like virus infectious to and replicable in B. mori-derived cells. PMID- 15827171 TI - H-2 Kd-restricted hepatitis B virus-derived epitope whose specific CD8+ T lymphocytes can produce gamma interferon without cytotoxicity. AB - It is necessary to evaluate the cytokine secretion status of CD8+ T lymphocytes and elucidate the factors influencing cytokine secretion, because the secretion of cytokines is also an important feature of CD8+ T lymphocytes, and the cytokines usually play critical roles in the outcome of diseases. We showed here that peptide AYRPPNAPI, derived from the core antigen of hepatitis B virus (HBV), could bind to H-2 Kd and induce primed splenocytes from HBcAg expression plasmid immunized mice to produce gamma interferon (IFN-gamma) in H-2 Kd- and CD8 dependent manners instead of in a CD4-dependent manner. The induced cells were mainly CD3 and CD8 positive but had no cytotoxic effect on the corresponding target cells. When administered into HBV transgenic mice, these cells can decrease the serum HBV load without causing liver damage. These results suggest that this peptide is a special kind of CD8+ T-cell epitope, for which specific CD8+ T cells can produce IFN-gamma when antigenic stimulation is encountered but which have no cytotoxic effect on the corresponding target cells both in vitro and in HBV transgenic mice. This phenomenon indicates initially that the functional mechanisms of CD8+ T cells can be determined by their epitope specificity, which may be associated with the development of epitope-based immunotherapeutic approaches for infectious diseases and tumors. PMID- 15827173 TI - Synthesis, assembly, and processing of the Env ERVWE1/syncytin human endogenous retroviral envelope. AB - Syncytin is a fusogenic protein involved in the formation of the placental syncytiotrophoblast layer. This protein is encoded by the envelope gene of the ERVWE1 proviral locus belonging to the human endogenous retrovirus W (HERV-W) family. The HERV-W infectious ancestor entered the primate lineage 25 to 40 million years ago. Although the syncytin fusion property has been clearly demonstrated, little is known about this cellular protein maturation process with respect to classical infectious retrovirus envelope proteins. Here we show that the cellular syncytin protein is synthesized as a glycosylated gPr73 precursor cleaved into two mature proteins, a gp50 surface subunit (SU) and a gp24 transmembrane subunit (TM). These SU and TM subunits are found associated as homotrimers. The intracytoplasmic tail is critical to the fusogenic phenotype, although its cleavage requirements seem to have diverged from those of classical retroviral maturation. PMID- 15827174 TI - Recruitment of CBP/p300, TATA-binding protein, and S8 to distinct regions at the N terminus of adenovirus E1A. AB - The N-terminal region of the adenovirus (Ad) 12S E1A gene product targets several cellular proteins that are essential for the induction of S phase, cellular immortalization, cellular transformation, transcriptional repression, and transcriptional activation. The precise binding sites for these proteins, however, remain to be resolved. We therefore undertook an extensive site-directed mutagenesis approach to generate specific point mutants and to precisely map the binding sites for CBP, p300, TATA-binding protein (TBP), S4, S8, hGcn5, P/CAF, and Ran within the first 30 amino acids of the Ad5 12S E1A protein. We determined that although common residues within the N-terminal region can form partial binding sites for these proteins, point mutants were also generated that could discriminate between binding sites. These data indicate that AdE1A can target each of these proteins individually through distinct binding sites. It was evident, however, that the mutation of specific hydrophobic residues typically had the greatest effect upon AdE1A's ability to bind individual partners. Indeed, the mutation of L at positions 19 and 20 eliminated the ability of AdE1A to interact with any of the N-terminal binding proteins studied here. Interestingly, although TBP and S8 or CBP/p300 can exist as functional complexes, RNA interference revealed that the recruitment of either TBP, S8, or CBP/p300 to AdE1A was not dependent upon the expression of the other proteins. These data further indicate that AdE1A can target individual partner proteins in vivo and that it does not necessarily recruit these proteins indirectly as components of larger macromolecular complexes. Finally, we took advantage of the fine-mapping data to ascertain which proteins were targeted during the transformation process. Consistent with previous studies, CBP/p300 was found to be targeted by AdE1A during this process, although our data suggest that binding to other N-terminal proteins is also important for transformation. PMID- 15827175 TI - Rift valley fever virus nonstructural protein NSs promotes viral RNA replication and transcription in a minigenome system. AB - Rift Valley fever virus (RVFV), which belongs to the genus Phlebovirus, family Bunyaviridae, has a tripartite negative-strand genome (S, M, and L segments) and is an important mosquito-borne pathogen for domestic animals and humans. We established an RVFV T7 RNA polymerase-driven minigenome system in which T7 RNA polymerase from an expression plasmid drove expression of RNA transcripts for viral proteins and minigenome RNA transcripts carrying a reporter gene between both termini of the M RNA segment in 293T cells. Like other viruses of the Bunyaviridae family, replication and transcription of the RVFV minigenome required expression of viral N and L proteins. Unexpectedly, the coexpression of an RVFV nonstructural protein, NSs, with N and L proteins resulted in a significant enhancement of minigenome RNA replication. Coexpression of NSs protein with N and L proteins also enhanced minigenome mRNA transcription in the cells expressing viral-sense minigenome RNA transcripts. NSs protein expression increased the RNA replication of minigenomes that originated from S and L RNA segments. Enhancement of minigenome RNA synthesis by NSs protein occurred in cells lacking alpha/beta interferon (IFN-alpha/beta) genes, indicating that the effect of NSs protein on minigenome RNA replication was unrelated to a putative NSs protein-induced inhibition of IFN-alpha/beta production. Our finding that RVFV NSs protein augmented minigenome RNA synthesis was in sharp contrast to reports that Bunyamwera virus (genus Bunyavirus) NSs protein inhibits viral minigenome RNA synthesis, suggesting that RVFV NSs protein and Bunyamwera virus NSs protein have distinctly different biological roles in viral RNA synthesis. PMID- 15827176 TI - An unrelated monoclonal antibody neutralizes human immunodeficiency virus type 1 by binding to an artificial epitope engineered in a functionally neutral region of the viral envelope glycoproteins. AB - Neutralizing antibodies often recognize regions of viral envelope glycoproteins that play a role in receptor binding or other aspects of virus entry. To address whether this is a necessary feature of a neutralizing antibody, we identified the V4 region of the gp120 envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) as a sequence that is tolerant of drastic change and thus appears to play a negligible role in envelope glycoprotein function. An artificial epitope tag was inserted into the V4 region without a significant effect on virus entry or neutralization by antibodies that recognize HIV-1 envelope glycoprotein sequences. An antibody directed against the artificial epitope tag was able to neutralize the modified, but not the wild-type, HIV-1. Thus, the specific target of a neutralizing antibody need not contribute functionally to the process of virus entry. PMID- 15827177 TI - Macrophage inflammatory protein 1alpha inhibits postentry steps of human immunodeficiency virus type 1 infection via suppression of intracellular cyclic AMP. AB - Primary isolates of human immunodeficiency virus type 1 (HIV-1) predominantly use chemokine receptor CCR5 to enter target cells. The natural ligands of CCR5, the beta-chemokines macrophage inflammatory protein 1alpha (MIP-1alpha), MIP-1beta, and RANTES, interfere with HIV-1 binding to CCR5 receptors and decrease the amount of virions entering cells. Although the inhibition of HIV-1 entry by beta chemokines is well documented, their effects on postentry steps of the viral life cycle and on host cell components that control the outcome of infection after viral entry are not well defined. Here, we show that all three beta-chemokines, and MIP-1alpha in particular, inhibit postentry steps of the HIV-1 life cycle in primary lymphocytes, presumably via suppression of intracellular levels of cyclic AMP (cAMP). Productive HIV-1 infection of primary lymphocytes requires cellular activation. Cell activation increases intracellular cAMP, which is required for efficient synthesis of proviral DNA during early steps of viral infection. Binding of MIP-1alpha to cognate receptors decreases activation-induced intracellular cAMP levels through the activation of inhibitory G proteins. Furthermore, inhibition of one of the downstream targets of cAMP, cAMP-dependent PKA, significantly inhibits synthesis of HIV-1-specific DNA without affecting virus entry. These data reveal that beta-chemokine-mediated inhibition of virus replication in primary lymphocytes combines inhibitory effects at the entry and postentry levels and imply the involvement of beta-chemokine-induced signaling in postentry inhibition of HIV-1 infection. PMID- 15827178 TI - Peptide mimetics of gamma interferon possess antiviral properties against vaccinia virus and other viruses in the presence of poxvirus B8R protein. AB - We have developed peptide mimetics of gamma interferon (IFN-gamma) that play a direct role in the activation and nuclear translocation of STAT1alpha transcription factor. These mimetics do not act through recognition by the extracellular domain of IFN-gamma receptor but rather bind to the cytoplasmic domain of the receptor chain 1, IFNGR-1, and thereby initiate the cellular signaling. Thus, we hypothesized that these mimetics would bypass the poxvirus virulence factor B8R protein that binds to intact IFN-gamma and prevents its interaction with the receptor. Human and murine IFN-gamma mimetic peptides were introduced into an adenoviral vector for intracellular expression. Murine IFN gamma mimetic peptide was also expressed via chemical synthesis with an attached lipophilic group for penetration of cell plasma membrane. In contrast to intact human IFN-gamma, the mimetics did not bind poxvirus B8R protein, a homolog of the IFN-gamma receptor extracellular domain. Expression of B8R protein in WISH cells did not block the antiviral effect of the mimetics against encephalomyocarditis or vesicular stomatitis virus, while the antiviral activity of human IFN-gamma was neutralized. Consistent with the antiviral activity, the upregulation of MHC class I molecules on WISH cells by the IFN-gamma mimetics was not affected by B8R protein, while IFN-gamma-induced upregulation was blocked. Finally, the mimetics, but not IFN-gamma, inhibited vaccinia virus replication in African green monkey kidney BSC-40 cells. The data presented demonstrate that small peptide mimetics of IFN-gamma can avoid the B8R virulence factor for poxviruses and, thus, are potential candidates for antivirals against smallpox virus. PMID- 15827179 TI - Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 replication and transcription activator regulates viral and cellular genes via interferon stimulated response elements. AB - Kaposi's sarcoma-associated herpesvirus (also called human herpesvirus 8 [HHV-8]) replication and transcription activator (RTA) is apparently necessary and sufficient for the switch from viral latency to lytic replication. RTA may regulate open reading frame (ORF) K14 (viral OX-2 homologue) and ORF74 (viral G protein-coupled receptor homologue) genes through an interferon-stimulated response element (ISRE)-like sequence (K14 ISRE) in the promoter region. RTA strongly activated a K14 ISRE-containing K14-ORF74 promoter reporter construct and a heterologous promoter reporter construct containing K14 ISRE. RTA could bind to K14 ISRE and other ISREs, activate promoter reporter constructs from interferon-simulated genes (ISGs), and selectively induce three endogenous ISGs in primary endothelial cells: ISG-54, myxovirus resistance protein 1 (MxA), and stimulated trans-acting factor of 50 kDa. In addition, a region in the RTA DNA binding domain has been identified with certain sequence similarity to the DNA binding domains of the interferon regulatory factor (IRF) family. Mutation in one conserved amino acid within this region reduced the ability of RTA to bind to ISRE as well as other RTA response elements. Furthermore, the mutant failed to activate RTA-responsive promoters and to induce viral lytic gene expression. The mutation at the same conserved amino acid residue in IRF-7 drastically reduced its ability to bind to DNA and to activate the beta interferon promoter. The sequence and functional similarities between RTA and IRFs suggest that the HHV-8 RTA may usurp the cellular IRF pathway. PMID- 15827180 TI - Evolution of the equine infectious anemia virus long terminal repeat during the alteration of cell tropism. AB - Equine infectious anemia virus (EIAV) is a lentivirus with in vivo cell tropism primarily for tissue macrophages; however, in vitro the virus can be adapted to fibroblasts and other cell types. Tropism adaptation is associated with both envelope and long terminal repeat (LTR) changes, and findings strongly suggest that these regions of the genome influence cell tropism and virulence. Furthermore, high levels of genetic variation have been well documented in both of these genomic regions. However, specific EIAV nucleotide or amino acid changes that are responsible for cell tropism changes have not been identified. A study was undertaken with the highly virulent, macrophage-tropic strain of virus EIAV(wyo) to identify LTR changes associated with alterations in cell tropism. We found the stepwise generation of a new transcription factor binding motif within the enhancer that was associated with adaptation of EIAV to endothelial cells and fibroblasts. An LTR that contained the new motif had enhanced transcriptional activity in fibroblasts, whereas the new site did not alter LTR activity in a macrophage cell line. This finding supports a previous prediction that selection for new LTR genetic variants may be a consequence of cell-specific selective pressures. Additional investigations of the EIAV(wyo) LTR were performed in vivo to determine if LTR evolution could be detected over the course of a 3-year infection. Consistent with previous in vivo findings, we observed no changes in the enhancer region of the LTR over that time period, indicating that the EIAV(wyo) LTR was evolutionarily stable in vivo. PMID- 15827181 TI - Early T-cell responses to dengue virus epitopes in Vietnamese adults with secondary dengue virus infections. AB - T-cell responses to dengue viruses may be important in both protective immunity and pathogenesis. This study of 48 Vietnamese adults with secondary dengue virus infections defined the breadth and magnitude of peripheral T-cell responses to 260 overlapping peptide antigens derived from a dengue virus serotype 2 (DV2) isolate. Forty-seven different peptides evoked significant gamma interferon enzyme-linked immunospot (ELISPOT) assay responses in 39 patients; of these, 34 peptides contained potentially novel T-cell epitopes. NS3 and particularly NS3200 324 were important T-cell targets. The breadth and magnitude of ELISPOT responses to DV2 peptides were independent of the infecting dengue virus serotype, suggesting that cross-reactive T cells dominate the acute response during secondary infection. Acute ELISPOT responses were weakly correlated with the extent of hemoconcentration in individual patients but not with the nadir of thrombocytopenia or overall clinical disease grade. NS3556-564 and Env414-422 were identified as novel HLA-A*24 and B*07-restricted CD8+ T-cell epitopes, respectively. Acute T-cell responses to natural variants of Env414-422 and NS3556 564 were largely cross-reactive and peaked during disease convalescence. The results highlight the importance of NS3 and cross-reactive T cells during acute secondary infection but suggest that the overall breadth and magnitude of the T cell response is not significantly related to clinical disease grade. PMID- 15827182 TI - Regulation of translation by ribosome shunting through phosphotyrosine-dependent coupling of adenovirus protein 100k to viral mRNAs. AB - Adenovirus simultaneously inhibits cap-dependent host cell mRNA translation while promoting the translation of its late viral mRNAs during infection. Studies previously demonstrated that tyrosine kinase activity plays a central role in the control of late adenovirus protein synthesis. The tyrosine kinase inhibitor genistein decreases late viral mRNA translation and prevents viral inhibition of cellular protein synthesis. Adenovirus protein 100k blocks cellular mRNA translation by disrupting the cap-initiation complex and promotes viral mRNA translation through an alternate mechanism known as ribosome shunting. 100k protein interaction with initiation factor eIF4G and the viral 5' noncoding region on viral late mRNAs, known as the tripartite leader, are both essential for ribosome shunting. We show that adenovirus protein 100k promotes ribosome shunting in a tyrosine phosphorylation-dependent manner. The primary sites of phosphorylated tyrosine on protein 100k were mapped and mutated, and two key sites are shown to be essential for protein 100k to promote ribosome shunting. Mutation of the two tyrosine phosphorylation sites in 100k protein does not impair interaction with initiation factor 4G, but it severely reduces association of 100k with tripartite leader mRNAs. 100k protein therefore promotes ribosome shunting and selective translation of viral mRNAs by binding specifically to the adenovirus tripartite leader in a phosphotyrosine-dependent manner. PMID- 15827183 TI - Memory T-cell response to rotavirus detected with a gamma interferon enzyme linked immunospot assay. AB - Measurements of serum-neutralizing antibody and anti-rotavirus immunoglobulin A (IgA) are the current standard for assessing immune responses following rotavirus vaccination. However, there is ongoing debate as to whether antibody titers correlate with protection against rotavirus gastroenteritis. Children recovering from rotavirus gastroenteritis have increased gamma interferon release from cultured peripheral blood mononuclear cells (PBMCs), suggesting that cell mediated immunity (CMI) may play a role in viral clearance and protection from subsequent gastroenteritis. We have developed a gamma interferon enzyme-linked immunospot (ELISPOT) assay for evaluation of CMI responses to rotavirus using frozen PBMCs obtained from healthy adults. Responses to three different rotavirus antigen types were analyzed-a peptide pool based on the human VP6 sequence; reassortant human:bovine vaccine strains; and cell culture-adapted (CCA) human G1, G2, G3, G4, and bovine (WC3) G6 strains. The reassortant strains consist of a bovine WC3 genome background expressing the human rotavirus surface proteins VP7 (G1, G2, G3, or G4) or VP4 (P1). Responses to titrations of the peptide pool as well as CCA and reassortant strains were assessed. Gamma interferon ELISPOT responses were similar for CCA and reassortant strains, whether live or UV inactivated, and when tested either individually or pooled. For most subjects, responses to the VP6 peptide pool positively correlated with responses to CCA and reassortant strains. Cell depletion studies indicate the memory responses detected with these frozen adult PBMCs were primarily due to the CD4+ T-cell population. This gamma interferon ELISPOT assay provides a new tool to apply in clinical studies for the characterization of natural or vaccine-induced CMI to rotavirus. PMID- 15827184 TI - Cell cycle arrest in G2/M promotes early steps of infection by human immunodeficiency virus. AB - We have identified four small molecules that boost transduction of cells by human immunodeficiency virus (HIV) and investigated their mechanism of action. These molecules include etoposide and camptothecin, which induce DNA damage by inhibiting religation of cleaved topoisomerase-DNA complexes, taxol, which interferes with the function of microtubules, and aphidicolin, which inhibits DNA polymerases. All four compounds arrest the cell cycle at G2/M, though in addition high concentrations of aphidicolin arrest in G1. We find that early events of HIV replication, including synthesis of late reverse transcription products, two-long terminal repeat circles, and integrated proviruses, were increased after treatment of cells with concentrations of each compound that arrested in G2/M. Stimulation was seen for both transformed cell lines (293T and HeLa cells) and primary cells (IMR90 lung fibroblasts). Arrest in G1 with high concentrations of aphidicolin boosted transduction, though not much as with lower concentrations that arrested in G2/M. Arrest of IMR90 cells in G1 by serum starvation and contact inhibition reduced transduction. Previously, the proteasome inhibitor MG132 was reported to increase HIV infection-here we investigated the effects of combinations of the cell cycle inhibitors with MG132 and obtained data suggesting that MG132 may also boost transduction by causing G2/M cell cycle arrest. These data document that cell cycle arrest in G2/M boosts the early steps of HIV infection and suggests methods for increasing transduction with HIV-based vectors. PMID- 15827185 TI - Inhibition of lysosome and proteasome function enhances human immunodeficiency virus type 1 infection. AB - We previously reported that inhibition of endosomal/lysosomal function can dramatically enhance human immunodeficiency virus type 1 (HIV-1) infectivity, suggesting that under these conditions productive HIV-1 infection can occur via the endocytic pathway. Here we further examined this effect with bafilomycin A1 (BFLA-1) and show that this enhancement of infectivity extends to all HIV-1 isolates tested regardless of coreceptor usage. However, isolate-specific differences were observed in the magnitude of the effect. This was particularly evident in the case of the weakly infectious HIV-1(SF2), for which we observed the greatest enhancement. Using reciprocal chimeric viruses, we were able to determine that both the disproportionate increase in the infectivity of HIV 1(SF2) in response to BFLA-1 and its weak infectivity in the absence of BFLA-1 mapped to its envelope gene. Further, we found HIV-1(SF2) to have lower fusion activity and to be 12-fold more sensitive to the fusion inhibitor T-20 than HIV 1(NL4-3). Proteasomal inhibitors also enhance HIV-1 infectivity, and we report that the combination of a lysosomal and a proteasomal inhibitor greatly enhanced infectivity of all isolates tested. Again, HIV-1(SF2) was unique in exhibiting a synergistic 400-fold increase in infectivity. We also determined that inhibition of proteasomal function increased the infectivity of HIV-1 pseudotyped with vesicular stomatitis virus G protein. The evidence presented here highlights the important role of the lysosomes/proteasomes in the destruction of infectious HIV 1(SF2) and could have implications for the development of novel antiviral agents that might take advantage of these innate defenses. PMID- 15827186 TI - T-cell growth transformation by herpesvirus saimiri is independent of STAT3 activation. AB - Herpesvirus saimiri (saimirine herpesvirus 2) (HVS), a T-lymphotropic tumor virus, induces lymphoproliferative disease in several species of New World primates. In addition, strains of HVS subgroup C are able to transform T cells of Old World primates, including humans, to permanently growing T-cell lines. In concert with the Stp oncoprotein, the tyrosine kinase-interacting protein (Tip) of HVS C488 is required for T-cell transformation in vitro and lymphoma induction in vivo. Tip was previously shown to interact with the protein tyrosine kinase Lck. Constitutive activation of signal transducers and activators of transcription (STATs) has been associated with oncogenesis and has also been detected in HVS-transformed T-cell lines. Furthermore, Tip contains a putative consensus YXPQ binding motif for the SH2 (src homology 2) domains of STAT1 and STAT3. Tip tyrosine phosphorylation at this site was required for binding of STATs and induction of STAT-dependent transcription. Here we sought to address the relevance of STAT activation for transformation of human T cells by introducing a tyrosine-to-phenylalanine mutation in the YXPQ motif of Tip of HVS C488. Unexpectedly, the recombinant virus was still able to transform human T lymphocytes, but it had lost its capability to activate STAT3 as well as STAT1. This demonstrates that growth transformation by HVS is independent of STAT3 activation. PMID- 15827187 TI - Rapid viral escape at an immunodominant simian-human immunodeficiency virus cytotoxic T-lymphocyte epitope exacts a dramatic fitness cost. AB - Escape from specific T-cell responses contributes to the progression of human immunodeficiency virus type 1 (HIV-1) infection. T-cell escape viral variants are retained following HIV-1 transmission between major histocompatibility complex (MHC)-matched individuals. However, reversion to wild type can occur following transmission to MHC-mismatched hosts in the absence of cytotoxic T-lymphocyte (CTL) pressure, due to the reduced fitness of the escape mutant virus. We estimated both the strength of immune selection and the fitness cost of escape variants by studying the rates of T-cell escape and reversion in pigtail macaques. Near-complete replacement of wild-type with T-cell escape viral variants at an immunodominant simian immunodeficiency virus Gag epitope KP9 occurred rapidly (over 7 days) following infection of pigtail macaques with SHIVSF162P3. Another challenge virus, SHIVmn229, previously serially passaged through pigtail macaques, contained a KP9 escape mutation in 40/44 clones sequenced from the challenge stock. When six KP9-responding animals were infected with this virus, the escape mutation was maintained. By contrast, in animals not responding to KP9, rapid reversion of the K165R mutation occurred over 2 weeks after infection. The rapidity of reversion to the wild-type sequence suggests a significant fitness cost of the T-cell escape mutant. Quantifying both the selection pressure exerted by CTL and the fitness costs of escape mutation has important implications for the development of CTL-based vaccine strategies. PMID- 15827188 TI - Homologous crossovers among molecules of brome mosaic bromovirus RNA1 or RNA2 segments in vivo. AB - Previously we demonstrated frequent homologous crossovers among molecules of the RNA3 segment in the tripartite brome mosaic bromovirus (BMV) RNA genome (A. Bruyere, M. Wantroba, S. Flasinski, A. Dzianott, and J. J. Bujarski, J. Virol. 74:4214-4219, 2000). To further our knowledge about mechanisms of viral RNA genome variability, in this paper we have studied homologous recombination in BMV RNA1 and RNA2 components during infection. We have found that basal RNA-RNA crossovers could occur within coding regions of both RNAs, although recombination frequencies slightly varied at different RNA sections. In all cases, the frequencies were much lower than the rate observed for the intercistronic recombination hot spot in BMV RNA3. Probability calculations accounted for at least one homologous crossover per RNA molecule per replication cycle. In addition, we have demonstrated an efficient repair of mutations within the conserved 3' and 5' noncoding regions, most likely due to error-prone BMV RNA replication. Overall, our data verify that homologous crossovers are common events a during virus life cycle, and we discuss their importance for viral RNA genetics. PMID- 15827189 TI - Evaluation of the conformational switch model for alfalfa mosaic virus RNA replication. AB - Key elements of the conformational switch model describing regulation of alfalfa mosaic virus (AMV) replication (R. C. Olsthoorn, S. Mertens, F. T. Brederode, and J. F. Bol, EMBO J. 18:4856-4864, 1999) have been tested using biochemical assays and functional studies in nontransgenic protoplasts. Although comparative sequence analysis suggests that the 3' untranslated regions of AMV and ilarvirus RNAs have the potential to fold into pseudoknots, we were unable to confirm that a proposed pseudoknot forms or has a functional role in regulating coat protein RNA binding or viral RNA replication. Published work has suggested that the pseudoknot is part of a tRNA-like structure (TLS); however, we argue that the canonical sequence and functional features that define the TLS are absent. We suggest here that the absence of the TLS correlates directly with the distinctive requirement for coat protein to activate replication in these viruses. Experimental data are evidence that elevated magnesium concentrations proposed to stabilize the pseudoknot structure do not block coat protein binding. Additionally, covarying nucleotide changes proposed to reestablish pseudoknot pairings do not rescue replication. Furthermore, as described in the accompanying paper (L. M. Guogas, S. M. Laforest, and L. Gehrke, J. Virol. 79:5752-5761, 2005), coat protein is not, by definition, inhibitory to minus-strand RNA synthesis. Rather, the activation of viral RNA replication by coat protein is shown to be concentration dependent. We describe the 3' organization model as an alternate model of AMV replication that offers an improved fit to the available data. PMID- 15827190 TI - Coat protein activation of alfalfa mosaic virus replication is concentration dependent. AB - Alfalfa mosaic virus (AMV) and ilarvirus RNAs are infectious only in the presence of the viral coat protein; therefore, an understanding of coat protein's function is important for defining viral replication mechanisms. Based on in vitro replication experiments, the conformational switch model states that AMV coat protein blocks minus-strand RNA synthesis (R. C. Olsthoorn, S. Mertens, F. T. Brederode, and J. F. Bol, EMBO J. 18:4856-4864, 1999), while another report states that coat protein present in an inoculum is required to permit minus strand synthesis (L. Neeleman and J. F. Bol, Virology 254:324-333, 1999). Here, we report on experiments that address these contrasting results with a goal of defining coat protein's function in the earliest stages of AMV replication. To detect coat-protein-activated AMV RNA replication, we designed and characterized a subgenomic luciferase reporter construct. We demonstrate that activation of viral RNA replication by coat protein is concentration dependent; that is, replication was strongly stimulated at low coat protein concentrations but decreased progressively at higher concentrations. Genomic RNA3 mutations preventing coat protein mRNA translation or disrupting coat protein's RNA binding domain diminished replication. The data indicate that RNA binding and an ongoing supply of coat protein are required to initiate replication on progeny genomic RNA transcripts. The data do not support the conformational switch model's claim that coat protein inhibits the initial stages of viral RNA replication. Replication activation may correlate with low local coat protein concentrations and low coat protein occupancy on the multiple binding sites present in the 3' untranslated regions of the viral RNAs. PMID- 15827191 TI - Binding and transfer of human immunodeficiency virus by DC-SIGN+ cells in human rectal mucosa. AB - The role of DC-SIGN on human rectal mucosal dendritic cells is unknown. Using highly purified human rectal mucosal DC-SIGN+ cells and an ultrasensitive real time reverse transcription-PCR assay to quantify virus binding, we found that HLA DR+/DC-SIGN+ cells can bind and transfer more virus than the HLA-DR+/DC-SIGN- cells. Greater than 90% of the virus bound to total mucosal mononuclear cells (MMCs) was accounted for by the DC-SIGN+ cells, which comprise only 1 to 5% of total MMCs. Significantly, anti-DC-SIGN antibodies blocked 90% of the virus binding when more-physiologic amounts of virus inoculum were used. DC-SIGN expression in the rectal mucosa was significantly correlated with the interleukin 10 (IL-10)/IL-12 ratio (r = 0.58, P < 0.002; n = 26) among human immunodeficiency virus (HIV)-positive patients. Ex vivo and in vitro data implicate the role of IL 10 in upregulating DC-SIGN expression and downregulating expression of the costimulatory molecules CD80/CD86. Dendritic cells derived from monocytes (MDDCs) in the presence of IL-10 render the MDDCs less responsive to maturation stimuli, such as lipopolysaccharide and tumor necrosis factor alpha, and migration to the CCR7 ligand macrophage inflammatory protein 3beta. Thus, an increased IL-10 environment could render DC-SIGN(+) cells less immunostimulatory and migratory, thereby dampening an effective immune response. DC-SIGN and the IL-10/IL-12 axis may play significant roles in the mucosal transmission and pathogenesis of HIV type 1. PMID- 15827192 TI - Scavenger receptor class B type I and hepatitis C virus infection of primary tupaia hepatocytes. AB - Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide. The study of early steps during HCV infection has been hampered by the lack of suitable in vitro or in vivo models. Primary Tupaia hepatocytes (PTH) have been shown to be susceptible to HCV infection in vitro and in vivo. Human scavenger receptor class B type I (SR-BI) represents an HCV receptor candidate mediating the cellular binding of E2 glycoprotein to HepG2 hepatoma cells. However, the function of SR-BI for viral infection of hepatocytes is unknown. In this study, we used PTH to assess the functional role of SR-BI as a putative HCV receptor. Sequence analysis of cloned tupaia SR-BI revealed a high homology between tupaia and human SR-BI. Transfection of CHO cells with human or tupaia SR-BI but not mouse SR-BI cDNA resulted in cellular E2 binding, suggesting that E2-binding domains between human and tupaia SR-BI are highly conserved. Preincubation of PTH with anti-SR-BI antibodies resulted in marked inhibition of E2 or HCV-like particle binding. However, anti-SR-BI antibodies were not able to block HCV infection of PTH. In conclusion, our results demonstrate that SR-BI represents an important cell surface molecule for the binding of the HCV envelope to hepatocytes and suggest that other or additional cell surface molecules are required for the initiation of HCV infection. Furthermore, the structural and functional similarities between human and tupaia SR-BI indicate that PTH represent a useful model system to characterize the molecular interaction of the HCV envelope and SR-BI on primary hepatocytes. PMID- 15827193 TI - Rhesus cytomegalovirus contains functional homologues of US2, US3, US6, and US11. AB - Human cytomegalovirus (HCMV) is a paradigm for mechanisms subverting antigen presentation by major histocompatibility complex (MHC) molecules. Due to its limited host range, HCMV cannot be studied in animals. Thus, the in vivo importance of inhibiting antigen presentation for the establishment and maintenance of infection with HCMV is unknown. Rhesus cytomegalovirus (RhCMV) is an emerging animal model that shares many of the features of HCMV infection. The recent completion of the genomic sequence of RhCMV revealed a significant degree of homology to HCMV. Strikingly, RhCMV contains several genes with low homology to the HCMV US6 gene family of inhibitors of the MHC I antigen presentation pathway. Here, we examine whether the RhCMV US6 homologues (open reading frames Rh182, -184, -185, -186, -187, and -189) interfere with the MHC I antigen processing pathway. We demonstrate that Rh182 and Rh189 function similarly to HCMV US2 and US11, respectively, mediating the proteasomal degradation of newly synthesized MHC I. The US3 homologue, Rh184, delayed MHC I maturation. Unlike US3, MHC I molecules eventually escaped retention by Rh184, so that steady-state surface levels of MHC I remained unchanged. Rh185 acted similarly to US6 and inhibited peptide transport by TAP and, consequently, peptide loading of MHC I molecules. Thus, despite relatively low sequence conservation, US6 family-related genes in RhCMV are functionally closely related to the conserved structural features of HCMV immunomodulators. The conservation of these mechanisms implies their importance for immune evasion in vivo, a question that can now be addressed experimentally. PMID- 15827194 TI - Tumorigenic poxviruses up-regulate intracellular superoxide to inhibit apoptosis and promote cell proliferation. AB - Tumorigenic leporipoxviruses encode catalytically inactive homologs of cellular Cu-Zn superoxide dismutase (SOD1). The function of the orthologous myxoma virus M131R and Shope fibroma virus S131R gene products is uncertain, but they inhibit SOD1 activity by a process linked to binding its copper chaperone. Using a superoxide-sensitive dye (hydroethidine), we observed that virus infection increased intracellular superoxide levels in an M/S131R-dependent manner. To see whether this effect promotes infection, we deleted the Shope fibroma virus S131R gene and compared the clinical manifestations of wild-type and mutant virus infections in rabbits. S131RDelta virus produced significantly smaller fibroxanthosarcoma-like growths in vivo and, at a point where these growths were already receding, wild-type infections still showed extensive leukocyte infiltration, necrosis, and fibromatous cell proliferation. Coincidentally, whereas Jurkat cells are protected from mitochondria- and Fas-mediated apoptosis by wild-type myxoma virus in vitro, M131RDelta virus could not block Fas initiated apoptosis as judged by DNA laddering, terminal deoxynucleotidyltransferase-mediated dUTP-fluorescein nick end labeling, and caspase 3 cleavage assays. These data suggest that tumorigenic poxviruses can modulate intracellular redox status to their advantage to stimulate infected cell growth and inhibit programmed cell death. PMID- 15827195 TI - Association of the influenza A virus RNA-dependent RNA polymerase with cellular RNA polymerase II. AB - Transcription by the influenza virus RNA-dependent RNA polymerase is dependent on cellular RNA processing activities that are known to be associated with cellular RNA polymerase II (Pol II) transcription, namely, capping and splicing. Therefore, it had been hypothesized that transcription by the viral RNA polymerase and Pol II might be functionally linked. Here, we demonstrate for the first time that the influenza virus RNA polymerase complex interacts with the large subunit of Pol II via its C-terminal domain. The viral polymerase binds hyperphosphorylated forms of Pol II, indicating that it targets actively transcribing Pol II. In addition, immunofluorescence analysis is consistent with a new model showing that influenza virus polymerase accumulates at Pol II transcription sites. The present findings provide a framework for further studies to elucidate the mechanistic principles of transcription by a viral RNA polymerase and have implications for the regulation of Pol II activities in infected cells. PMID- 15827196 TI - Effect of antiviral treatment with entecavir on age- and dose-related outcomes of duck hepatitis B virus infection. AB - Entecavir (ETV), a potent inhibitor of the hepadnaviral polymerases, prevented the development of persistent infection when administered in the early stages of duck hepatitis B virus (DHBV) infection. In a preliminary experiment, ETV treatment commenced 24 h before infection showed no significant advantage over simultaneous ETV treatment and infection. In two further experiments 14-day-old ducks were inoculated with DHBV-positive serum containing 10(4), 10(6), 10(8), or 5 x 10(8) viral genomes (vge) and were treated orally with 1.0 mg/kg of body weight/day of ETV for 14 or 49 days. A relationship between virus dose and infection outcome was seen: non-ETV-treated ducks inoculated with 10(4) vge had transient infection, while ducks inoculated with higher doses developed persistent infection. ETV treatment for 49 days did not prevent initial infection of the liver but restricted the spread of infection more than approximately 1,000 fold, a difference which persisted throughout treatment and for up to 49 days after withdrawal. Ultimately, three of seven ETV-treated ducks resolved their DHBV infection, while the remaining ducks developed viremia and persistent infection after a lag period of at least 63 days. ETV treatment for 14 days also restricted the spread of infection, leading to marked and sustained reductions in the number of DHBV-positive hepatocytes in 7 out of 10 ducks. In conclusion, short-term suppression with ETV provides opportunity for the immune response to successfully control DHBV infection. Since DHBV infection of ducks provides a good model system for HBV infection in humans, it seems likely that ETV may be useful in postexposure therapy for HBV infection aimed at preventing the development of persistent infection. PMID- 15827197 TI - Aged BALB/c mice as a model for increased severity of severe acute respiratory syndrome in elderly humans. AB - Advanced age has repeatedly been identified as an independent correlate of adverse outcome and a predictor of mortality in cases of severe acute respiratory syndrome (SARS). SARS-associated mortality may exceed 50% for persons aged 60 years or older. Heightened susceptibility of the elderly to severe SARS and the ability of SARS coronavirus to replicate in mice led us to examine whether aged mice might be susceptible to disease. We report here that viral replication in aged mice was associated with clinical illness and pneumonia, demonstrating an age-related susceptibility to SARS disease in animals that parallels the human experience. PMID- 15827198 TI - Endoplasmic reticulum-localized human papillomavirus type 16 E5 protein alters endosomal pH but not trans-Golgi pH. AB - The human papillomavirus type 16 (HPV-16) E5 protein is a small, hydrophobic polypeptide that is expressed in virus-infected keratinocytes and alters receptor signaling pathways, apoptotic responses, and endosomal pH. Despite its ability to inhibit endosomal acidification, the HPV-16 E5 protein is found predominantly in the endoplasmic reticulum (ER), suggesting that its effect may be indirect and perhaps global. To determine whether E5 alters the pHs of additional intracellular compartments, we transduced human keratinocytes with a codon optimized E5 vector and then quantified endosomal and trans-Golgi pHs using sensitive, compartment-specific, ratiometric pHluorin constructs. E5 protein increased endosomal pH from 5.9 to 6.9 but did not affect the normal trans-Golgi pH of 6.3. Confirming the lack of alteration in trans-Golgi pH, we observed no alterations in the acidification-dependent processing of the proH3 protein. C terminal deletions of E5, which retained normal expression and localization in the ER, were defective for endosomal alkalization. Thus, E5 does not uniformly alkalinize intracellular compartments, and its C-terminal 10 amino acids appear to mediate interactions with critical ER targets that modulate proton pump function and/or localization. PMID- 15827199 TI - Pathological prion protein in the tongues of sheep infected with naturally occurring scrapie. AB - Tongue involvement by prion spreading was shown to be a common outcome after oral or intracranial experimental challenge with scrapie and transmissible mink encephalopathy sources in rodent models. It is also known that bovine spongiform encephalopathy, which is pathogenic for humans, is experimentally transmissible to sheep and can lead to a disease indistinguishable from scrapie. A recent European Food Safety Authority opinion recommended research into PrPsc accumulation in the tongues of ruminants. We report on the detection of PrPsc in the tongues of seven scrapie-infected sheep by immunohistochemistry and Western blotting. PMID- 15827200 TI - Identification of functional domains in kaposica, the complement control protein homolog of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8). AB - Recently it has been shown that kaposica, an immune evasion protein of Kaposi's sarcoma-associated herpesvirus, inactivates complement by acting on C3 convertases by accelerating their decay as well as by acting as a cofactor in factor I-mediated inactivation of their subunits C3b and C4b. Here, we have mapped the functional domains of kaposica. We show that SCRs 1 and 2 (SCRs 1-2) and 1-4 are essential for the classical and alternative pathway C3-convertase decay-accelerating activity (DAA), respectively, while the SCRs 2-3 are required for factor I cofactor activity (CFA) for C3b and C4b. SCR 3 and SCRs 1 and 4, however, contribute to optimal classical pathway DAA and C3b CFA, respectively. Binding data show that SCRs 1-4 and SCRs 1-2 are the smallest structural units required for measuring detectable binding to C3b and C4b, respectively. The heparin-binding site maps to SCR 1. PMID- 15827201 TI - Nearby clusters of hemagglutinin residues sustain SLAM-dependent canine distemper virus entry in peripheral blood mononuclear cells. AB - Signaling lymphocytic activation molecule (SLAM, CD150) is the universal morbillivirus receptor. Based on the identification of measles virus (MV) hemagglutinin (H) amino acids supporting human SLAM-dependent cell entry, we mutated canine distemper virus (CDV) H and identified residues necessary for efficient canine SLAM-dependent membrane fusion. These residues are located in two nearby clusters in a new CDV H structural model. To completely abolish SLAM dependent fusion, combinations of mutations were necessary. We rescued a SLAM blind recombinant CDV with six mutations that did not infect ferret peripheral blood mononuclear cells while retaining full infectivity in epithelial cells. PMID- 15827202 TI - Characterization of herpes simplex virus type 1 thymidine kinase mutants selected under a single round of high-dose brivudin. AB - A broad variety of herpes simplex virus type 1 clones was selected under a single round of high-dose selection with brivudin. Mutations in the thymidine kinase (TK) genes consisted of 42% frameshift mutations within homopolymer repeats of G's and C's and single nucleotide substitutions (58%) that produced stop codons (Q261 and R281) or a new codon at the site of the substitution (A168T, R51W, G59W, G206R, R220H, Y239S, and T287 M). The A168T change, associated with an altered TK phenotype, proved to be the most commonly selected substitution. For the different mutants, a correlation between phenotype, genotype, and in vivo neurovirulence was observed. PMID- 15827203 TI - Viral RNA is required for the association of APOBEC3G with human immunodeficiency virus type 1 nucleoprotein complexes. AB - APOBEC3G (APO3G) is a host cytidine deaminase that is incorporated into human immunodeficiency virus type 1 (HIV-1) particles. We report here that viral RNA promotes stable association of APO3G with HIV-1 nucleoprotein complexes (NPC). A target sequence located within the 5'-untranslated region of the HIV-1 RNA was identified to be necessary and sufficient for efficient APO3G packaging. Fine mapping revealed a sequence normally involved in viral genomic RNA dimerization and Gag binding to be important for APO3G packaging and association with viral NPC. Our data suggest that packaging of APO3G into HIV-1 NPC is enhanced by viral RNA. PMID- 15827204 TI - Induction of lytic Epstein-Barr virus (EBV) infection by synergistic action of rituximab and dexamethasone renders EBV-positive lymphoma cells more susceptible to ganciclovir cytotoxicity in vitro and in vivo. AB - The purposeful induction of the lytic form of Epstein-Barr virus (EBV) infection combined with ganciclovir (GCV) treatment has been advocated as a novel strategy for EBV-positive B-cell lymphoma. We demonstrated that rituximab had a synergistic effect with dexamethasone on induction of the lytic EBV infection in CD20-positive lymphoma cells. Addition of GCV to the dexamethasone/rituximab treated cells was more effective than dexamethasone/rituximab alone in killing EBV-positive lymphoma cells in vitro and in lymphoma-bearing nude mice but not in EBV-negative cells. These data suggest that induction of the lytic EBV infection with dexamethasone/rituximab in combination with GCV could be a potential virally targeted therapy for EBV-associated B-cell lymphoma. PMID- 15827205 TI - Hyperphosphorylation of EBNA2 by Epstein-Barr virus protein kinase suppresses transactivation of the LMP1 promoter. AB - The Epstein-Barr virus (EBV) BGLF4 gene encodes a serine/threonine protein kinase (PK) that is expressed in the cytolytic cycle. EBV nuclear antigen 2 (EBNA2) is a key latency gene essential for immortalization of B lymphocytes and transactivation of viral and cellular promoters. Here we report that EBV PK phosphorylates EBNA2 at Ser-243 and that these two proteins physically associate. PK suppresses EBNA2's ability to transactivate the LMP1 promoter, and Ser-243 of EBNA2 is involved in this suppression. Moreover, EBNA2 is hyperphosphorylated during EBV reactivation in latently infected B cells, which is associated with decreased LMP1 protein levels. This is the first report about the effect of EBV PK on the function of one of its target proteins and regulation of EBNA2 phosphorylation during the EBV lytic cycle. PMID- 15827207 TI - Population dynamics of a diverse rodent assemblage in mixed grass-shrub habitat, southeastern Colorado, 1995-2000. AB - We followed seasonal and year-to-year population dynamics for a diverse rodent assemblage in a short-grass prairie ecosystem in southeastern Colorado (USA) for 6 yr. We captured 2,798 individual rodents (range, one to 812 individuals per species) belonging to 19 species. The two most common species, deer mice (Peromyscus maniculatus) and western harvest mice (Reithrodontomys megalotis), generally had population peaks in winter and nadirs in summer; several other murid species demonstrated autumn peaks and spring nadirs; heteromyids were infrequently captured in winter, and populations generally peaked in summer or autumn. Inter-annual trends indicated an interactive effect between temperature and precipitation. Conditions associated with low rodent populations or population declines were high precipitation during cold periods (autumn and winter) and low precipitation during warm periods (spring and summer). Severity of adverse effects varied by species. Heteromyids, for example, were apparently not negatively affected by the hot, dry spring and summer of 2000. Cross correlations for the temporal series of relative population abundances between species pairs (which are affected by both seasonal and interannual population dynamics) revealed positive associations among most murids and among most heteromyids, but there were negative associations between murids and heteromyids. These results have important implications for those attempting to model population dynamics of rodent populations for purposes of predicting disease risk. PMID- 15827206 TI - Epizootiology of Sin Nombre and El Moro Canyon hantaviruses, southeastern Colorado, 1995-2000. AB - Sin Nombre virus (SNV) is an etiologic agent of hantavirus pulmonary syndrome. To better understand the natural history of this virus we studied population dynamics and temporal pattern of infection of its rodent hosts in southeastern Colorado (USA) from 1995 to 2000. We present evidence for the presence of two hantaviruses, SNV in deer mice (Peromyscus maniculatus) and El Moro Canyon virus in western harvest mice (Reithrodontomys megalotis), at our study sites. Sin Nombre virus appeared only sporadically in deer mouse populations; overall prevalence of antibody to SNV was 2.6%. El Moro Canyon virus was enzootic: seroconversions occurred throughout the year; antibody prevalence (11.9% overall) showed a delayed-density-dependent pattern, peaking as relative abundance of mice was declining. Males of both host species were more frequently infected than were females. An apparently lower mean survivorship (persistence at the trapping site) for SNV antibody-positive deer mice could indicate a detrimental effect of SNV on its host, but might also be explained by the fact that antibody-positive mice were older when first captured. PMID- 15827208 TI - Fibropapillomatosis in stranded green turtles (Chelonia mydas) from the eastern United States (1980-98): trends and associations with environmental factors. AB - We examined data collected by the US Sea Turtle Stranding and Salvage Network on 4,328 green turtles (Chelonia mydas) found dead or debilitated (i.e., stranded) in the eastern half of the USA from Massachusetts to Texas during the period extending from 1980 to 1998. Fibropapillomatosis (FP) was reported only on green turtles in the southern half of Florida (south of 29 degrees N latitude). Within this region, 22.6% (682/3,016) of the turtles had tumors. Fibropapillomatosis was more prevalent in turtles found along the western (Gulf) coast of Florida (51.9%) than in turtles found along the eastern (Atlantic) coast of Florida (11.9%) and was more prevalent in turtles found in inshore areas (38.9%) than in turtles found in offshore areas (14.6%). A high prevalence of FP corresponded to coastal waters characterized by habitat degradation and pollution, a large extent of shallow-water area, and low wave energy, supporting speculation that one or more of these factors could serve as an environmental cofactor in the expression of FP. A high prevalence of FP did not correspond to high-density green turtle assemblages. Turtles with tumors were found most commonly during the fall and winter months, and the occurrence of tumors was most common in turtles of intermediate size (40-70-cm curved carapace length). Stranded green turtles with tumors were more likely to be emaciated or entangled in fishing line and less likely to have propeller wounds than were stranded green turtles without tumors. Turtles with and without tumors were equally likely to show evidence of a shark attack. The percent occurrence of tumors in stranded green turtles increased from approximately 10% in the early 1980s to over 30% in the late 1990s. Fibropapillomatosis was first documented in southernmost Florida in the late 1930s and spread throughout the southern half of Florida and the Caribbean during the mid-1980s. Because green turtles living in south Florida are known to move throughout much of the Caribbean, but are not known to move to other parts of the USA or to Bermuda, the spread and current distribution of FP in the western Atlantic, Gulf of Mexico, and Caribbean can be explained by assuming FP is caused by an infectious agent that first appeared in southern Florida. Aberrant movements of captive-reared turtles or of turtles that are released into areas where they were not originally found could spread FP beyond its current distribution. PMID- 15827209 TI - European brown hare syndrome in free-living mountain hares (Lepus timidus) and European brown hares (Lepus europaeus) in Finland 1990-2002. AB - We investigated the epidemiology of the European brown hare syndrome (EBHS) in Finland among free-living mountain hares (Lepus timidus) and European brown hares (Lepus europaeus). Carcasses of 297 European brown hares and 843 mountain hares submitted for postmortem examination were analyzed between 1990 and 2002. One hundred thirty-six cases of EBHS were detected during the examination, 20 cases in European brown hares and 116 in mountain hares. Liver samples from 56 histopathologically positive and 135 negative cases were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The results of RT-PCR correlated well with histopathology. Cases of EBHS appeared only sporadically in a restricted area in eastern Finland during 1990-97. In 1998, the number of cases increased, and the disease spread to the north. In 1998-99, EBHS was the most common disease of mountain hares in necropsy material from northern and eastern Finland. Special features were the occurrence of the disease in areas in which European brown hares are rare and the seasonal distribution of cases, most of which appeared in spring or summer. This is the first report of an outbreak of EBHS occurring mainly among free-living mountain hares from Finland. PMID- 15827210 TI - Avian cholera in waterfowl: the role of lesser snow and ross's geese as disease carriers in the Playa Lakes Region. AB - We collected samples from apparently healthy geese in the Playa Lakes Region (USA) during the winters of 2000-01 and 2001-02 to determine whether carriers of Pasteurella multocida, the bacterium that causes avian cholera, were present in wild populations. With the use of methods developed in laboratory challenge trials (Samuel et al., 2003a) and a serotype-specific polymerase chain reaction method for identification of P. multocida serotype 1, we found that a small proportion of 322 wild birds (<5%) were carriers of pathogenic P. multocida. On the basis of serology, an additional group of these birds (<10%) were survivors of recent avian cholera infection. Our results confirm the hypothesis that wild waterfowl are carriers of avian cholera and add support for the hypothesis that wild birds are a reservoir for this disease. In concert with other research, this work indicates that enzootic infection with avian cholera occurs in lesser snow goose (Chen caerulescens caerulescens) populations throughout their annual cycle. Although fewer Ross's geese (Chen rossii) were sampled, we also found these birds were carriers of P. multocida. Even in the absence of disease outbreaks, serologic evidence indicates that chronic disease transmission and recent infection are apparently occurring year-round in these highly gregarious birds and that a small portion of these populations are potential carriers with active infection. PMID- 15827211 TI - Epizootiologic investigations of selected infectious disease agents in free ranging Eurasian lynx from Sweden. AB - Serum samples from 106 Eurasian lynx (Lynx lynx) from across Sweden, found dead or shot by hunters in 1993-99, were investigated for presence of antibodies to feline parvovirus (FPV), feline coronavirus, feline calicivirus, feline herpesvirus, feline immunodeficiency virus, Francisella tularensis, and Anaplasma phagocytophila, and for feline leukemia virus antigen. In addition, tissue samples from 22 lynx submitted in 1999 were analyzed by real-time polymerase chain reaction (PCR) to detect nucleic acids specific for viral agents and A. phagocytophila. Except for FPV antibodies in one lynx and A. phagocytophila in four lynx, all serology was negative. All PCR results also were negative. It was concluded that free-ranging Swedish lynx do not have frequent contact with the infectious agents considered in this study. PMID- 15827212 TI - Serologic surveillance for selected viral agents in captive and free-ranging populations of Arabian oryx (Oryx leucoryx) from Saudi Arabia and the United Arab Emirates. AB - A total of 294 sera collected between 1999 and 2001 from eight captive and one free-ranging herds of Arabian oryx (Oryx leucoryx) distributed in Saudi Arabia (SA) and the United Arab Emirates (UAE) were assayed for antibodies against 13 selected viral agents. Arabian oryx have been exposed to bluetongue virus (BTV), epizootic hemorrhagic disease virus (EHDV), rinderpest virus (RPV), bovine respiratory syncytial virus (BRSV), bovine adenovirus 3 (BAV-3), cervid herpesvirus-1, foot-and-mouth disease virus, equine herpesvirus 9, and bovine viral diarrhea virus. The high seroprevalence to BTV and EHDV in the UAE and SA indicates that Arabian oryx are likely to be susceptible to infection by these viruses and therefore could act as a source of virus to vectors during the infective stage of infection. Moreover, antibodies were detected against RPV and BRSV in sera from SA and against BAV-3 in sera from the UAE. No antibodies were found against bovine herpesvirus-1, caprine herpesvirus-1, enzootic bovine leucosis virus, and peste des petits ruminants virus. On the basis of these results, caution should be applied when considering translocation of Arabian oryx, and only those proven to be free of infectious agents that might present a risk to other species should be moved. PMID- 15827213 TI - Antibodies against equine herpesviruses and equine arteritis virus in Burchell's zebras (Equus burchelli ) from the Serengeti ecosystem. AB - A total of 51 sera from a migratory population of Burchell's zebras (Equus burchelli) were collected in the Serengeti National Park (Tanzania) between 1999 and 2001 to assess levels of exposure to equine herpesvirus types 1, 2, 4, 9 (EHV 1, -2, -4, -9), EHV-1 zebra isolate T965, and equine arteritis virus (EAV). Using virus-specific neutralizing antibody tests, seroprevalence was high for EHV-9 (60% of 45), moderate for EAV (24% of 51), and lower for the EHV-1-related zebra isolate (17% of 41), EHV-1 (14% of 49), and EHV-4 (2% of 50). No evidence for exposure to EHV-2 was found (0% of 51). The high level of exposure to EHV-9 is interesting because evidence of infection with this virus has not been previously described in any wild equine population. Although the epidemiology of EHV-9 in Burchell's zebras is presently unknown, our results suggest that in East Africa, this species may be a natural host of EHV-9, a neuropathogenic virus that was only recently isolated from captive Thomson's gazelles (Gazella thomsoni) in Japan. There is currently no evidence that EHV-9 induced mortality in Burchell's zebras in the Serengeti, but because of the reported virulence of this virus for more susceptible species such as Thomson's gazelles, viral transmission from infected zebras to ungulates may result in mortality. PMID- 15827214 TI - Anesthesia and blood sampling of wild big brown bats (eptesicus fuscus) with an assessment of impacts on survival. AB - We anesthetized and blood sampled wild big brown bats (Eptesicus fuscus) in Fort Collins, Colorado (USA) in 2001 and 2002 and assessed effects on survival. Inhalant anesthesia was delivered into a specially designed restraint and inhalation capsule that minimized handling and bite exposures. Bats were immobilized an average of 9.1+/-5.1 (SD) min (range 1-71, n=876); blood sample volumes averaged 58+/-12 microl (range 13-126, n=718). We randomly selected control (subject to multiple procedures before release) and treatment (control procedures plus inhalant anesthesia and 1% of body weight blood sampling) groups in 2002 to assess treatment effects on daily survival over a 14-day period for adult female and volant juvenile bats captured at maternity roosts in buildings. We monitored survival after release using passive integrated transponder tag detection hoops placed at openings to selected roosts. Annual return rates of bats sampled in 2001 were used to assess long-term outcomes. Comparison of 14-day maximum-likelihood daily survival estimates from control (86 adult females, 92 volant juveniles) and treated bats (187 adult females, 87 volant juveniles) indicated no adverse effect from anesthesia and blood sampling (juveniles: chi2=22.22, df=27, P>0.05; adults: chi2=9.72, df=18, P>0.05). One-year return rates were similar among adult female controls (81%, n=72, 95% confidence interval [CI]=70-91%), females treated once (82%, n=276, 95% CI=81-84%), and females treated twice (84%, n=50, 95% CI=74-94%). Lack of an effect was also noted in 1-yr return rates of juvenile female controls (55%, n=29, 95% CI=37 73%), juveniles treated once (66%, n=113, 95% CI=58-75%), and juveniles treated twice (71%, n=17, 95% CI=49-92%). These data suggest that anesthesia and blood sampling for health monitoring did not measurably affect survival of adult female and volant juvenile big brown bats. PMID- 15827215 TI - West Nile virus infection in farmed American alligators (Alligator mississippiensis) in Florida. AB - In September and October 2002, an epizootic of neurologic disease occurred at an alligator farm in Florida (USA). Three affected American alligators (Alligator mississippiensis) were euthanatized and necropsied, and results confirmed infection with West Nile virus (WNV). The most significant microscopic lesions were a moderate heterophilic to lymphoplasmacytic meningoencephalomyelitis, necrotizing hepatitis and splenitis, pancreatic necrosis, myocardial degeneration with necrosis, mild interstitial pneumonia, heterophilic necrotizing stomatitis, and glossitis. Immunohistochemistry identified WNV antigen, with the most intense staining in liver, pancreas, spleen, and brain. Virus isolation and RNA detection by reverse transcription-polymerase chain reaction confirmed WNV infection in plasma and tissue samples. Of the tissues, liver had the highest viral loads (maximum 10(8.9) plaque-forming units [PFU]/0.5 cm3), whereas brain and spinal cord had the lowest viral loads (maximum 10(6.6) PFU/0.5 cm3 each). Virus titers in plasma ranged from 10(3.6) to 10(6.5) PFU/ml, exceeding the threshold needed to infect Culex quinquefasciatus mosquitoes (10(5) PFU/ml). Thus, alligators may serve as a vertebrate amplifying host for WNV. PMID- 15827216 TI - Validation and use of an indirect enzyme-linked immunosorbent assay for detection of antibodies to West Nile virus in American Alligators (Alligator mississippiensis) in Florida. AB - In October 2002, West Nile virus (WNV) was identified in farmed American alligators (Alligator mississippiensis) in Florida showing clinical signs and having microscopic lesions indicative of central nervous system disease. To perform seroepidemiologic studies, an indirect enzyme-linked immunosorbent assay (ELISA) was developed to determine exposure of captive and wild alligators to WNV. To validate the test, a group of WNV-seropositive and -seronegative alligators were identified at the affected farm using hemagglutination inhibition (HAI) and the plaque reduction neutralization test (PRNT). The indirect ELISA utilized a rabbit anti-alligator immunoglobulins polyclonal antibody as the secondary antibody, and inactivated WNV-infected Vero cells were used as the coating antigen. For all samples (n=58), the results of the ELISA were consistent with the HAI and PRNT findings. Plasma was collected from 669 free-ranging alligators from 21 sites across Florida in April and October 2003. Four samples collected in April and six in October were positive for WNV antibodies using HAI, PRNT, and the indirect ELISA. This indicated that wild alligators in Florida have been exposed to WNV. These findings can be used as a baseline for future surveys. PMID- 15827218 TI - A comparison of four serologic assays in screening for Brucella exposure in Hawaiian monk seals. AB - A survey for Brucella spp. antibodies was undertaken on 164 serum samples from 144 Hawaiian monk seals (Monachus schauinslandi) from the northwestern Hawaiian Islands collected between 1995 and 2002. The buffered antigen plate agglutination test (BPAT), the indirect enzyme immunoassay (I-ELISA), the competitive enzyme immunoassay (C-ELISA), and the fluorescence polarization assay (FPA) were compared with regard to their ability in detecting antibodies to Brucella spp. in the serum samples. Overall, antibodies were detected in 28 (17.1%) animals, using the BPAT test, 25 (15.2%) by the C-ELISA, and 19 (11.6%) in the I-ELISA and the FPA test, using thresholds established for cattle. No evidence of gross pathology consistent with clinical brucellosis was noted in any of the seropositive animals tested. Although further work would be necessary to validate these tests for use with monk seals it appears that both the C-ELISA and the FPA tests would be appropriate as diagnostic screening tests for detection of antibodies to Brucella spp. in this species. PMID- 15827217 TI - Columbian black-tailed deer (Odocoileus hemionus columbianus) as hosts for Borrelia spp. in northern California. AB - The prevalence of infection of Columbian black-tailed deer (Odocoileus hemionus columbianus) with Borrelia spp. was evaluated in an area of northwestern California (USA) where Lyme disease is endemic and the relapsing-fever group spirochete Borrelia coriaceae is enzootic, and in a far-removed comparison area having a disparate climate and lower density of vector ticks. Blood samples collected from both deer herds in 1987, 1988, and from 2000-02 were assayed for borrelial infection with microscopic and molecular methods. Serum specimens from two (5%) of 39 deer from the Dye Creek Preserve in Tehama County versus 13 (20%) of 64 animals from the Hopland Research and Extension Center (HREC) in Mendocino County, California were polymerase chain reaction (PCR) test positive for B. burgdorferi sensu lato. DNA sequencing analyses revealed that eight animals were infected with B. bissettii, six with three unclassified genotypes, and one with B. burgdorferi sensu stricto. One serum sample (2%) from HREC was positive for a relapsing-fever group spirochete that had a 16S rRNA sequence homology of 99% with the C053 type strain of B. coriaceae. Spirochetes undetermined to geno species were detected in thick-blood drops prepared from three (8%) of 36 deer from the HREC by direct immunofluorescence. Adults of the hippoboscid flies Lipoptena depressa (n=73) and Neolipoptena ferrisi (n=24), the Pacific Coast tick (Dermacentor occidentalis) (n=22), and the western black-legged tick (Ixodes pacificus) (n=1) that had been removed from deer from both study areas in 2002 were PCR test negative for borreliae. The occurrence of diverse borreliae in deer from northern California confounds and, consequently, reduces the utility of borrelial serosurveys for detecting specific genospecies, unless they are complemented by more specific assays (e.g., immunoblotting, PCR/sequencing analysis). PMID- 15827219 TI - Mortality of Rocky Mountain elk in Michigan due to meningeal worm. AB - Mortality from cerebrospinal parelaphostrongylosis caused by the meningeal worm (Parelaphostrongylus tenuis) has been hypothesized to limit elk (Cervus elaphus nelsoni) populations in areas where elk are conspecific with white-tailed deer (Odocoileus virginianus). Elk were reintroduced into Michigan (USA) in the early 1900s and subsequently greatly increased population size and distribution despite sympatric high-density (>or=12/km2) white-tailed deer populations. We monitored 100 radio-collared elk of all age and sex classes from 1981-94, during which time we documented 76 mortalities. Meningeal worm was a minor mortality factor for elk in Michigan and accounted for only 3% of mortalities, fewer than legal harvest (58%), illegal kills (22%), other diseases (7%), and malnutrition (4%). Across years, annual cause-specific mortality rates due to cerebrospinal parelaphostrongylosis were 0.033 (SE=0.006), 0.029 (SE=0.005), 0.000 (SE=0.000), and 0.000 (SE=0.000) for calves, 1-yr-old, 2-yr-old, and >or=3-yr-old, respectively. The overall population-level mortality rate due to cerebrospinal parelaphostrongylosis was 0.009 (SE=0.001). Thus, meningeal worm had little impact on elk in Michigan during our study despite greater than normal precipitation (favoring gastropods) and record (>or=14 km2) deer densities. Further, elk in Michigan have shown sustained population rates-of-increase of >or=18%/yr and among the highest levels of juvenile production and survival recorded for elk in North America, indicating that elk can persist in areas with meningeal worm at high levels of population productivity. It is likely that local ecologic characteristics among elk, white-tailed deer, and gastropods, and degree of exposure, age of elk, individual and population experience with meningeal worm, overall population vigor, and moisture determine the effects of meningeal worm on elk populations. PMID- 15827220 TI - Interactive effects of turkeypox virus and Plasmodium hermani on Turkey poults. AB - Two experiments were conducted to examine the interactive effects of two disease agents of wild turkeys (Meleagris gallopavo), turkeypox virus and the malarial organism, Plasmodium hermani, on the health of turkey poults. Groups of domestic broad-breasted white turkey poults of 1 and 10 wk of age were infected with either turkeypox virus, P. hermani, both turkeypox virus and P. hermani, or were maintained as uninfected controls. The strains of turkeypox virus and P. hermani had been isolated from wild turkeys in southern Florida (USA). The goals of these experiments were two-fold and included both an examination of age differences in response to infections, and an examination of the effects of dual versus singular infections with the two agents. Both singular and concomitant infections of turkeypox virus and P. hermani were more detrimental to poults infected at 1 wk of age than to those infected at 10 wk, based on mortality, weight gain, and parasitemia. Dual infections of turkeypox virus and P. hermani were found to be slightly more harmful to 1-wk-old poults than were singular infections. No such interactive effects were noted in the poults infected at 10 wk of age. PMID- 15827221 TI - Helminths of saiga antelope in Kazakhstan: implications for conservation and livestock production. AB - Saiga antelope (Saiga tatarica) graze extensively on livestock pasture, potentially enabling transmission of a wide range of parasitic helminths between saigas and domestic ruminants. Thirty-six of the 38 species of helminth that have been found in saigas in Kazakhstan in the past have been found also in domestic livestock. We examined 133 saigas culled for meat in autumn 1997, and found three species of cestode and 12 nematodes (nine in the abomasum), but no trematodes or lungworms. The most abundant species were Marshallagia marshalli, Marshallagia mongolica, and Nematodirus gazellae in the abomasum, Nematodirus gazellae in the small intestine, and Skrjabinema ovis in the large intestine. There was no clear relationship between intensities of abomasal nematodes and body condition. Age intensity patterns differed between species: N. gazellae intensities were highest in saigas around 2-3 yr old, and declined in older animals, whereas the intensity of Marshallagia spp. rose asymptotically with age. Fecal egg density was directly proportional to adult worm intensity across ages for Marshallagia spp., but only in young animals for N. gazellae. There was no evidence that helminths, at the intensities observed, adversely affect saiga populations. The host range of many of the parasites found is broad, and transmission between saigas and livestock in both directions might become important to agriculture and conservation as livestock numbers recover. Simplified sampling techniques used in this study, and statistical analysis based on bootstrapping, could prove useful in other parasitologic surveys of wildlife in remote areas. PMID- 15827222 TI - Attempts to identify the source of avian vacuolar myelinopathy for waterbirds. AB - Attempts were made to reproduce avian vacuolar myelinopathy (AVM) in a number of test animals in order to determine the source of the causative agent for birds and to find a suitable animal model for future studies. Submerged vegetation, plankton, invertebrates, forage fish, and sediments were collected from three lakes with ongoing outbreaks of AVM and fed to American coots (Fulica americana), mallard ducks and ducklings (Anas platyrhynchos), quail (Coturnix japonica), and laboratory mice either via gavage or ad libitum. Tissues from AVM-affected coots with brain lesions were fed to ducklings, kestrels (Falco sparverius), and American crows (Corvus brachyrhynchos). Two mallards that ingested one sample of Hydrilla verticillata along with any biotic or abiotic material associated with its external surface developed brain lesions consistent with AVM, although neither of the ducks had clinical signs of disease. Ingestion of numerous other samples of Hydrilla from the AVM affected lakes and a lake with no prior history of AVM, other materials (sediments, algae, fish, invertebrates, and water from affected lakes), or tissues from AVM-affected birds did not produce either clinical signs or brain lesions in any of the other test animals in our studies. These results suggest that waterbirds are most likely exposed to the causative agent of AVM while feeding on aquatic vegetation, but we do not believe the vegetation itself is the agent. We hypothesize that the causative agent of AVM might either be accumulated by aquatic vegetation, such as Hydrilla, or associated with biotic or abiotic material on its external surfaces. In support of that hypothesis, two coots that ingested Hydrilla sampled from a lake with an ongoing AVM outbreak in wild birds developed neurologic signs within 9 days (ataxia, limb weakness, and incoordination), and one of two coots that ingested Hydrilla collected from the same site 13 days later became sick and died within 38 days. None of these three sick coots had definitive brain lesions consistent with AVM by light microscopy, but they had no gross or histologic lesions in other tissues. It is unclear if these birds died of AVM. Perhaps they did not ingest a dose sufficient to produce brain lesions or the lesions were ultrastructural. Alternatively, it is possible that a separate neurotoxic agent is responsible for the morbidity and mortality observed in these coots. PMID- 15827223 TI - Birth, morphologic, and blood characteristics of free-ranging white-tailed deer neonates. AB - Winter severity is a primary factor influencing deer survival and reproduction in northern climates. Prolonged, harsh winters can adversely affect body condition of does, resulting in depressed morphologic development of neonates. In this study, we captured 59 white-tailed deer (Odocoileus virginianus) neonates (28 in 2001 and 31 in 2002), following two distinctly different winters, one severe and the other historically mild. Vaginal implant transmitters allowed exact age to be determined for 73% of the neonates; new hoof growth was used to estimate age (days) of the other 27%. Birthdate and morphologic measurements of neonates (i.e., birth mass, new hoof growth, hoof length) were compared by sex and capture year. For known-age neonates (n=43), there was a year-by-sex interaction effect (P=0.01) on birthdate, being later for females during spring 2001 compared with 2002, which was consistent with a significant (P=0.03) year-by-sex interaction for total hoof length (22.3 mm [SE=0.9] and 20.3 [SE=0.8] for females and males in 2001; 19.9 [SE=1.0] and 22.1 [SE=1.0] for females and males in 2002). Interestingly, there was no effect of year on birth mass or birthdate of known age neonates. A year-by-sex interaction (P=0.04) was determined for birthdates of estimated age (5 yr old were born later (P<0.01) than fawns born to dams 32 raccoons within the KLWR's known range on Key Largo, Florida, USA. All samples were negative for eggs of B. procyonis. Raccoon density in this area was approximately 0.62 raccoons/ha. Despite this high density of raccoons, B. procyonis does not appear to be a threat to the KLWR population. PMID- 15827233 TI - Lead poisoning in whooper and tundra swans. AB - Six weak whooper swans (Cygnus cygnus) and two weak tundra swans (Cygnus columbianus) were found at Swamp Miyajima (Hokkaido, Japan) in May 1998. Anorexia, depression, green watery feces, pale conjunctiva, and anemia were observed. Radiographs showed from six to 38 suspected lead pellets in the gizzard. Blood lead concentrations were 2.5-6.7 microg/g (mean+/-SD=4.6+/-1.14 microg/g) on day 1. After blood collection, the birds were treated with calcium disodium ethylenediaminetetraacetate (CaEDTA) given intravenously and force fed. Despite treatment, seven birds died the next day. Green, bile-stained livers and pale or green kidneys were observed on necropsy. Microscopically, bile pigment was widespread in the liver and acid-fast intranuclear inclusion bodies were observed in renal tubular epithelium. Lead concentrations in livers and kidneys were 14.0-30.4 microg/g and 30.2-122 microg/g wet weight, respectively. Only one bird survived and this whooper swan continued to be treated with CaEDTA and activated charcoal. No lead shot was observed in the proventriculus and gizzard by radiography on day 64 and the blood lead concentration decreased from 2.9 microg/g to 0.09 microg/g during that same period. After 4 mo of rehabilitation, the whooper swan was returned to the wild. Lead intoxication continues to be a problem at Swamp Miyajima. PMID- 15827234 TI - New distribution records of Echinococcus multilocularis in the brown lemming from Barrow, Alaska, USA. AB - We identified Echinococcus multilocularis for the first time in brown lemmings (Lemmus trimucronatus) from Barrow, Alaska, USA. Of 467 brown lemmings trapped between 1995 and 2000, two males and two females (0.9%; 95% confidence interval=0.9+/-0.9%) were found to be infected with metacestodes of E. multilocularis. No metacestodes were found in 17 collared lemmings (Dicrostonyx rubricatus) also trapped at Barrow. In humans, E. multilocularis causes alveolar echinococcosis, which is potentially fatal. Knowledge of the distribution of this parasite is important to protect the public health. PMID- 15827235 TI - Molecular characterization of rat gastric mucosal response to potent acid inhibition. AB - Potent acid inhibition with proton pump inhibitors (PPIs) is widely used in clinical medicine, especially for gastroesophageal reflux disease. PPIs cause profound changes in the intragastric environment with near-neutral pH and increase serum concentration of the gastric secretagogue hormone gastrin. Long term hypergastrinemia increases mucosal thickness and enterochromaffin-like cell density in gastric corpus mucosa and results in development of gastric carcinoids in experimental animals. Our aim was to study responses to potent acid inhibition by characterizing genome-wide gene expression changes in gastric corpus mucosa in rats dosed with the PPI omeprazole. Nine rats received 400 micromol/kg omeprazole daily for 10 wk. Seven rats received vehicle only. Analysis of gastric corpus with microarrays representing 11,848 genes identified 134 genes with changed gene expression levels in omeprazole-dosed rats. Several of the identified genes were previously known to be affected by potent acid inhibition. Of the 62 genes with known functions that changed gene expression levels after PPI dosing, 27 are known to be involved in proliferation and apoptosis and immune, inflammatory, and stress responses. Our study indicates that microarray analysis can detect relevant gene expression changes in the complex gastric tissue, and that cellular processes involved in cell growth and defense responses are strongly affected by PPI dosing. Many genes are identified that were not previously known to be affected by inhibition of gastric acid secretion or that have unknown biological functions. Characterization of the roles of these genes may give new insight into molecular responses to treatment with PPIs. PMID- 15827236 TI - Maternal diet programs embryonic kidney gene expression. AB - Human epidemiological data associating birth weight with adult disease suggest that organogenesis is "programmed" by maternal diet. In rats, protein restriction in pregnancy produces offspring with fewer renal glomeruli and higher systemic blood pressures than controls. We tested the hypothesis that maternal diet alters gene expression in the metanephros, the precursor of the definitive mammalian kidney. We demonstrated that maternal low-protein diet initiated when pregnancy starts and maintained to embryonic day 13, when the metanephros consists of mesenchyme surrounding a once-branched ureteric bud, is sufficient to significantly reduce glomerular numbers in offspring by about 20%. As assessed by representational difference analyses and real-time quantitative polymerase chain reactions, low-protein diet modulated gene expression in embryonic day 13 metanephroi. In particular, levels of prox-1, the ortholog of Drosophila transcription factor prospero, and cofilin-1, a regulator of the actin cytoskeleton, were reduced. During normal metanephrogenesis, prox-1 protein was first detected in mesenchymal cells around the ureteric tree and thereafter in nascent nephron epithelia, whereas cofilin-1 immunolocalized to bud derivatives and condensing mesenchyme. Previously, we reported that low-protein diets increased mesenchymal apoptosis cells when metanephrogenesis began and thereafter reduced numbers of precursor cells. Collectively, these studies prove that the maternal diet programs the embryonic kidney, altering cell turnover and gene expression at a time when nephrons and glomeruli have yet to form. The human implication is that the maternal diet ingested between conception and 5- 6-wk gestation contributes to the variation in glomerular numbers that are known to occur between healthy and hypertensive populations. PMID- 15827237 TI - Macroarray analysis of coelomocyte gene expression in response to LPS in the sea urchin. Identification of unexpected immune diversity in an invertebrate. AB - The purple sea urchin, Strongylocentrotus purpuratus, is a member of the phylum Echinodermata, which is basal to the phylum Chordata within the deuterostome lineage of the animal kingdom. This relationship makes the analysis of the sea urchin immune system relevant to understanding the evolution of the deuterostome immune system leading to the Vertebrata. Subtractive suppression hybridization was employed to generate cDNA probes for screening high-density arrayed, conventional cDNA libraries to identify genes that were upregulated in coelomocytes responding to lipopolysaccharide. Results from 1,247 expressed sequence tags (ESTs) were used to infer that coelomocytes upregulated genes involved in RNA splicing, protein processing and targeting, secretion, endosomal activities, cell signaling, and alterations to the cytoskeletal architecture including interactions with the extracellular matrix. Of particular note was a set of transcripts represented by 60% of the ESTs analyzed, which encoded a previously uncharacterized family of closely related proteins, provisionally designated as 185/333. These transcripts exhibited a significant level of variation in their nucleotide sequence and evidence of putative alternative splicing that could yield up to 15 translatable elements. On the basis of the striking increase in gene expression in response to lipopolysaccharide and the unexpected level of diversity of the 185/333 messages, we propose that this set of transcripts encodes a family of putative immune response proteins that may represent a major component of an immunological response to bacterial challenge. PMID- 15827238 TI - Severe hypertension caused by alleles from normotensive Lewis for a quantitative trait locus on chromosome 2. AB - Pursuing fully a suggestion from linkage analysis that there might be a quantitative trait locus (QTL) for blood pressure (BP) in a chromosome (Chr) 2 region of the Dahl salt-sensitive rat (DSS), four congenic strains were made by replacing various fragments of DSS Chr 2 with those of Lewis (LEW). Consequently, a BP QTL was localized to a segment of around 3 cM or near 3 Mb on Chr 2 by comparative congenics. The BP-augmenting alleles of this QTL originated from the LEW rat, a normotensive strain compared with DSS. The dissection of a QTL with such a paradoxical effect illustrated the power of congenics in unearthing a gene hidden in the context of the whole animal system, presumably by interactions with other genes. The locus for the angiotensin II receptor AT-1B (Agtr1b) is not supported as a candidate gene for the QTL because a congenic strain harboring it did not have an effect on BP. There are approximately 19 known and unknown genes present in the QTL interval. Among them, no standout candidate genes are reputed to affect BP. Thus the QTL will likely represent a novel gene for BP regulation. PMID- 15827240 TI - Polymorphism of the DQA2 gene in goats. AB - Variation in the caprine DQA2 gene was investigated using PCR-single-strand conformational polymorphism (SSCP) and DNA sequencing. Eleven DQA2 alleles were defined by SSCP patterns from 23 goats. All the caprine alleles shared high sequence homology to ovine DQA2 sequences, and exhibited a pattern of polymorphism similar to DQA2 alleles from sheep and cattle but different from caprine DQA1 sequences. Thirty-eight AA positions in the alpha1 domain of caprine DQA2 molecules were polymorphic, and a high degree of polymorphism was observed in the putative antigen-binding region, with 74% of the positions being polymorphic. Phylogenetic analysis of caprine, ovine, and bovine DQA sequences revealed that the caprine DQA2 sequences identified here grouped with ovine DQA2, bovine DQA2, DQA3, and DQA4 sequences but are separate from the group of caprine DQA1 alleles. Nine of the caprine DQA2 sequences were more similar to ovine DQA2 alleles, whereas the remaining two were more closely related to ovine DQA2-like and bovine DQA3 alleles. This finding suggests that the caprine DQA2 sequences may represent two loci, which probably arose by either gene duplication or gene conversion events. Allelic lineages were evident for both DQA2 and DQA2-like loci, supporting the trans-species mode of evolution of major histocompatibilitly complex genes. The high level of polymorphism and similarity between caprine and ovine DQA2 alleles suggests that the DQA2 gene may play an important role in immune responses to shared pathogens. PMID- 15827241 TI - Identification of errors and factors associated with errors in data from electronic swine feeders. AB - Electronic swine feeders are used to automatically measure individual feed intake on group-housed pigs, but the resulting data contain errors caused by feeder malfunctions and animal-feeder interactions. The objectives of this study were to 1) develop criteria to identify errors in data from an electronic feeder that is predominant in the United States; 2) evaluate the frequency of errors in data from three consecutive experiments using the same feeders; and 3) identify factors associated with errors. Across experiments, data included 1,878,321 feed intake records (visits) on 1,721 pigs and 124 pens. Sixteen criteria were developed to detect errors in seven variables related to feed trough weights and times. Logistic regression was used to identify factors associated with the presence or absence of each error type in identified visits (visits where the feeder recognized a transponder) using a model that included the fixed effects of replicate, sex, linear and quadratic effects of day on test, and random effects of feeder within replicate, pig within feeder within replicate, test day within replicate, and week within feeder within replicate. Frequencies of error types in identified visits varied considerably within and between experiments. Errors in feed trough weights were more frequent than errors in time. Percentage of identified visits and of daily feed intake records with at least one error ranged from 4.3 to 18.7% and from 17.2 to 50.0%, respectively, and decreased from the first to the last experiment, reflecting the increasing ability of the managers to operate the feeders. Replicate, sex, test day, feeder within replicate, pig, and day within replicate affected the number of errors that occurred, but their effect varied among error types. Week-to-week variation within a feeder and replicate had the largest effect on number of errors, which was likely associated with feeder management. Results indicate that the frequency of errors in data from electronic swine feeders is substantial, but visits with errors can be identified and their frequency can be decreased by proper feeder management. PMID- 15827242 TI - Selection for placental efficiency in swine: genetic parameters and trends. AB - The objectives of this study were to estimate response to divergent selection for an index of placental efficiency in swine, and to evaluate the effect of placental efficiency on litter size. The selection index (SI) included total born (TB), birth weight (BRWT), and placental weight (PW), and was designed to increase in the high line (H) or decrease in the low line (L) the efficiency of the placental function (PE), defined as the ratio BRWT:PW. (Co)variance components were estimated for direct and maternal additive effects by using an animal model with MTDFREML procedures. Estimated breeding values were calculated by using records on individual BRWT (n = 2,111), PW (n = 2,006), PE (n = 1,677), and SI (n = 1,677). Litter traits were evaluated using records on 193 litters. The model included the fixed effects of contemporary group for all traits, with the addition of sex for individual traits and parity for litter traits. Litter was fitted as an uncorrelated random effect for all traits, and TB was used as a linear and quadratic covariate for BRWT, PW, and PE. Direct heritability estimates from single-trait models were 0.03, 0.25, 0.18, 0.11, and 0.08 for BRWT, PW, PE, SI, and TB, respectively. Estimated breeding values were compared between lines by using a model including generation, line within generation, and replicate within line as the error term. Estimates of genetic divergence were 20.7 +/- 2.7 g, 0.24 +/- 0.03, 0.11 +/- 0.02, and 0.07 +/- 0.02 per generation for PW, PE, SI, and TB, respectively (P < 0.01), but divergence was not significant for BRWT. At Generation 4, direct EBV was higher in L than in H for PW (55.9 +/- 8.7 vs. -24.2 +/- 9.5 g, respectively; P < 0.01) and higher in H than in L for PE (0.58 +/- 0.10 vs. -0.35 +/- 0.09 g, respectively; P < 0.01). However, EBV was not different for BRWT, SI, or TB. These results indicate that PW and PE are susceptible to change by genetic selection; however, the correlated response in TB was an unexpected genetic trend toward a higher TB in L of 0.05 +/ 0.01 piglets per generation (P < 0.01). PMID- 15827243 TI - Changes in inbreeding of U.S. Herefords during the twentieth century. AB - Genetic diversity in the U.S. Hereford population was characterized by examining the level and rate of inbreeding and effective population size. Pedigree records for 20,624,418 animals were obtained from the American Hereford Association, of which 96.1% had both parents identified. Inbreeding coefficients were computed and mean inbreeding (Fx) calculated by year from 1900 to 2001. Inbreeding increased rapidly between 1900 and 1945. From 1946, inbreeding increased linearly to a maximum of 11.5% in 1966. Throughout the 1970s and 1980s, mean inbreeding decreased to mid-century levels. Several alternatives were investigated to explain this decline. The average relationship between prominent sires fell from 20 to 12% during the time that the level of inbreeding decreased, which reflects an increase in the popularity of certain less fashionable sire lines that would have temporarily decreased inbreeding. Pedigrees were constructed for animals born after 1990. This subsample of animals with no missing ancestors in at least 12 generations did not exhibit a decrease in inbreeding. Missing ancestral information therefore contributed to the apparent decline. One cause of missing ancestry results from outcrossing to imported animals. The effect of missing ancestry was investigated by simulating the missing ancestors. In 2001, Fx was 9.8%, and approximately 95% of individuals were inbred. The maximal inbreeding coefficient was 76%. The annual change in mean inbreeding (DeltaFx) was estimated for Herefords born during five time periods from 1946 to 2001, where inbreeding was changing at different linear rates. The DeltaFx for the most recent generation (1990 to 2001) was 0.12%/yr. Assuming a generation interval of 4.88 yr, the estimated effective population size was 85. This study provides a benchmark of current genetic diversity in the Hereford population. Results indicate that inbreeding is accumulating linearly and below critical levels. Increases in the adoption of reproductive technologies could decrease genetic diversity, and in the future, we may need to consider strategies to minimize inbreeding. PMID- 15827244 TI - Pigs weaned from the sow at 10 days of age respond to dietary energy source of manufactured liquid diets and exogenous porcine somatotropin. AB - Previous research indicates that the neonatal pig does not alter feed intake in response to changes in the energy density of manufactured liquid diets. Also, the limited response of IGF-I to exogenous porcine ST (pST) previously observed in young pigs may be influenced by the source of dietary energy. Our objectives were to 1) determine the effect of a high-fat (HF; 25% fat and 4,639 kcal/kg ME; DM basis) or low-fat (LF; 2% fat and 3,481 kcal/kg ME; DM basis) manufactured liquid diet on pig performance; and 2) determine whether the limited response to exogenous pST in young pigs depends on the source of dietary energy. Two replicates of 60 pigs (n = 120; barrows and gilts distributed evenly), with an initial BW of 4,207 +/- 51 g, were weaned from the sow at 10 d of age and used in a randomized complete block design. Pigs were assigned by BW to one of six pens. Diets were formulated to provide a constant lysine:ME ratio and were fed on a pen basis for a duration of 9 d. On d 5, barrows and gilts within a pen were assigned randomly to receive either 0 or 120 microg of pST.kg BW(-1).d(-1) for 4 d. Pigs gained 336 +/- 9 g/d, which resulted in an ending BW of 7,228 +/- 120 g, regardless of dietary treatment (P > 0.15). Pigs fed the LF diet consumed 17% more DM per pen daily than pigs fed the HF diet (2,777 +/- 67 vs. 2,376 +/- 67 g/d, P < 0.01), but calculated ME intake did not differ between dietary treatments (P > 0.20). The G:F was 24% greater in HF- than in LF-fed pigs (P < 0.01). Plasma urea N concentrations were higher in the HF-fed pigs (11.0 +/- 0.6 mg/dL) than in pigs fed the LF diet (6.2 +/- 0.6 mg/dL; P < 0.05). Treatment with pST increased circulating IGF-I (P < 0.01) and decreased PUN (P < 0.01) concentration 32 and 25%, respectively, regardless of dietary treatment (P > 0.30). Circulating leptin averaged 1.8 +/- 0.1 ng/mL and was not affected by dietary treatment (P > 0.35) or pST (P > 0.40). These results suggest that the ST/IGF axis is responsive in the young pig and the increase in circulating IGF-I and growth is independent of the source of dietary energy. Also, young pigs respond to a lower energy density liquid diet with increased feed intake, without altering growth performance, apparently utilizing a mechanism other than circulating leptin. PMID- 15827245 TI - A method to establish co-cultures of myotubes and preadipocytes from collagenase digested neonatal pig semitendinosus muscles. AB - The relationships between adipocyte and muscle cell development within muscle are important in the study of factors or agents that may improve meat quality. Neonatal porcine muscle has the potential to yield both cell types for cell culture because it contains developing adipocytes and a high number of muscle satellite cells. Therefore, we modified a conventional collagenase-based procedure to digest neonatal porcine muscle and subsequently cultured the resultant muscle stromal-vascular (SV) cells on several substrata in basal and dexamethasone (DEX)-containing media. Developing myotubes and preadipocytes were present in muscle SV cell cultures on laminin substrata following seeding and plating with fetal bovine serum (FBS) with or without DEX. Myotube number was much higher (P < 0.05) on laminin substrata compared with all other substrata, whereas preadipocyte number in muscle SV cell cultures was independent of substrata, as we have shown previously. This approach can be used to establish co cultures of differentiating adipocytes and myotubes from collagenase-digested neonatal pig muscle. Because the comparison is within the same culture dish, this method allows for a direct comparison of the responses of adipogenic and myogenic cells to growth and differentiation factors. For example, DEX did not alter myogenesis (i.e., 11 +/- 3 vs. 11 +/- 4 myotubes per unit area for control and DEX-treated cultures, respectively), but it has been shown to markedly increase preadipocyte number in muscle SV cell cultures. PMID- 15827246 TI - Factors associated with early and mid-to-late fetal loss in lactating and nonlactating Holstein cattle in a hot climate. AB - The purpose of this study was to evaluate associations of lactation, somatic cell count score (SCCS) at breeding, milk yield, lactation number, interval from calving to breeding (days open), number of times inseminated, and season of breeding on fetal loss for lactating Holstein females (both first-parity and multiparous cows) and nonlactating Holstein heifers in a hot climate. Females were palpated between d 40 and 50 of gestation and again at d 70 to 80 to determine pregnancy status. Early fetal loss was defined as a loss that occurred after d 40 to 50 but before d 70 to 80. Mid-to-late fetal loss represented losses after d 70 to 80 but before expected calving. Lactating females had higher early (P = 0.055) and mid-to-late fetal loss (P < 0.05) than nonlactating heifers. Those lactating females with increased days open experienced greater early (P < 0.05) and mid-to-late fetal loss (P = 0.055), whereas lactating females with an elevated SCCS encountered greater mid-to-late fetal loss (P < 0.01). Milk yield, lactation number, number of times inseminated, and season were not associated with early or mid-to-late fetal loss. For nonlactating heifers, there were no associations between number of times inseminated, season, or age at breeding on early or mid-to-late fetal loss. In conclusion, lactating females were more likely to suffer early and mid-to-late fetal loss than nonlactating heifers. Also, days open and SCCS at breeding were related to ability of lactating females to maintain pregnancy, but there were no relationships between fetal loss and milk yield, lactation number, number of times inseminated, or season. PMID- 15827247 TI - Effects of short-term feed deprivation and melatonin implants on circadian patterns of leptin in the horse. AB - Leptin is a protein hormone produced by adipose tissue that influences hypothalamic mechanisms regulating appetite and energy balance. In species tested thus far, including horses, concentrations of leptin increase as animal fat mass increases. The variables and mechanisms that influence the secretion of leptin are not well known, nor is it known in equine species how the secretion of leptin is influenced by acute alterations in energy balance, circadian patterns, and/or reproductive competence. Our objectives were to determine in horses: 1) whether plasma concentrations of leptin are secreted in a circadian and/or a pulsatile pattern; 2) whether a 48-h period of feed restriction would alter plasma concentrations of leptin, growth hormone, or insulin; and 3) whether ovariectomy and/or a melatonin implant would affect leptin. In Exp. 1, mares exposed to ambient photoperiod of visible light (11 h, 33 min to 11 h, 38 min), received treatments consisting of a 48-h feed restriction (RES) or 48 h of alfalfa hay fed ad libitum (FED). Mares were maintained in a dry lot before sampling and were tethered to a rail during sampling. Analyses revealed that leptin was not secreted in a pulsatile manner, and that mean leptin concentrations were greater (P < 0.001) in FED vs. RES mares (17.20 +/- 0.41 vs. 7.29 +/- 0.41 ng/mL). Plasma growth hormone was pulsatile, and mean concentrations were greater in RES than FED mares (2.15 +/- 0.31 vs. 1.08 +/- 0.31 ng/mL; P = 0.05). Circadian patterns of leptin secretion were observed, but only in FED mares (15.39 +/- 0.58 ng/mL for morning vs. 19.00 +/- 0.58 ng/mL for evening; P < 0.001). In Exp. 2, mares that were ovariectomized or intact received either a s.c. melatonin implant or a sham implant. Thereafter, blood was sampled at weekly intervals at 1000 and 1700. Concentrations of leptin in samples collected at 1700 were greater (P < 0.001) than in those collected at 1000 (28.24 +/- 1.7 vs. 22.07 +/- 1.7 ng/mL). Neither ovariectomy nor chronic treatment with melatonin affected plasma concentrations of leptin or the circadian pattern of secretion. These data provide evidence that plasma leptin concentrations in the equine are sensitive to acute changes in nutritional status and vary in a circadian pattern that is sensitive to fasting but not to melatonin treatment or ovariectomy. PMID- 15827248 TI - Influence of starch intake on growth and skeletal development of weanling horses. AB - Forty-four weanling horses were used in two experiments to evaluate the effect of starch intake on growth and skeletal development. In Exp. 1, the weanlings were fed either a grain-based, high-starch (31.1%, DM basis) concentrate or a by product-based, low-starch (0.0%) concentrate with coastal bermudagrass (Cynodon dactylon) hay. Corn oil was used to equalize the energy concentration of the concentrates. The concentrate:hay ratio was 64:36 (as-fed basis), and intake was the same for both diets. Body weight gains were greater by the weanlings consuming the high-starch concentrate (0.81 vs. 0.67 kg/d; P = 0.01). Total body length gain also was greater for the weanlings consuming the high-starch concentrate (15.5 vs. 13.2 cm; P = 0.045). Other body measurements and bone mineral deposition were not influenced by diet or gender. At the end of the experiment, postprandial blood glucose concentrations suggested that the horses on the low-starch diet were less efficient in metabolizing blood glucose than were those that had been consuming the high-starch diets. In Exp. 2, the weanlings were fed either a high-starch (34.7%) or medium-starch (17.0%) concentrate plus coastal bermudagrass hay. Corn oil again was used to equalize the energy content of the medium-starch concentrate to that of the high-starch concentrate. The concentrate:hay ratio was 64:36 (as-fed basis), and the intake was the same for both diets. The diets did not influence rate of gain (0.75 kg/d; P = 0.98), body measurements (P = 0.11 to 0.93), or bone mineral deposition (P = 0.66). Animals on the medium-starch diet tended to have blood glucose concentrations that peaked earlier and were lower at later times than those consuming the high-starch concentrate. Bone osteochondrotic lesions were not related to the diet and were found to decrease during the course of the experiment for both the high-starch and the medium-starch diets (P = 0.006 and 0.016, respectively). PMID- 15827249 TI - Increasing dietary pectin level reduces utilization of digestible threonine intake, but not lysine intake, for body protein deposition in growing pigs. AB - Two N balance studies were conducted to investigate the effects of feeding graded levels of pectin (a soluble nonstarch polysaccharide, NSP) on the utilization of ileal digestible threonine (Thr; Thr study) and lysine (Lys; Lys study) intake for body protein deposition (PD) in growing pigs. In each study, eight Yorkshire barrows with an average initial BW of 17.2 +/- 1.3 (Thr study) and 14.3 +/- 1.4 kg (Lys study) were fed each of five experimental diets during five subsequent experimental periods, according to a crossover design. Pigs were fed twice daily at 2.6 times maintenance energy requirements. The soybean- and cornstarch-based diets, in which either Thr or Lys was the first-limiting nutrient, were formulated to contain (as-fed basis) 0, 4, 8, or 12% pectin or 8% cellulose (water-insoluble NSP), respectively, and with NSP substituting cornstarch. Across treatments, the mean daily Thr and Lys intake were 5.42 +/- 0.04 g/d (Thr study) and 7.98 +/- 0.12 g/d (Lys study), respectively. Apparent and standardized ileal digestibilities of Thr and Lys were determined in a separate study. Mean PD was 93.4, 90.2, 82.1, 76.7, and 87.9 g/d (SEM = 1.3; Thr study) and 90.7, 88.6, 87.8, 85.3, and 88.1 g/d (SEM = 1.1; Lys study) for the five respective treatments. Utilization of ileal digestible Thr intake, but not of ileal digestible Lys intake, for PD decreased linearly with dietary pectin level, and was not influenced by diet cellulose level. The current study indicates that apparent and standardized ileal digestibility values do not provide an accurate predictor of dietary effects on the utilization of ileal digestible Thr intake for protein deposition in growing pigs fed diets containing soluble NSP. PMID- 15827250 TI - Bioavailability of phosphorus in meat and bone meal for swine. AB - Meat and bone meal (MBM), when supplemented with tryptophan, is an excellent protein source for pigs. It is also a rich source of Ca and P, but some research has suggested that the bioavailability of P is variable. Experiment 1 further examined the bioavailability of P in MBM. The MBM was obtained directly from a plant and was processed to pass through a 10-mesh screen. It contained 50.7% CP, 2.26% lysine, 10.0% Ca, and 5.0% P (air-dry basis). Individually penned pigs (n = 35; 17 kg initial BW) were fed (ad libitum basis) a low-P, corn-soybean meal basal diet (0.95% lysine, 0.70% Ca, 0.34% P; as-fed basis) or the basal with graded levels of added P (0.067, 0.133, 0.200%) from monosodium phosphate (MSP) or MBM for 40 d. The Ca level was 0.70% in all diets. Diets were fortified with salt, vitamins, and trace minerals. At termination, the third and fourth metacarpals and metatarsals and femurs were removed from all pigs. Growth rate and feed:gain improved linearly (P < 0.01) with P addition, regardless of source, whereas ADFI was unaffected (P = 0.20). Bone strength and ash increased linearly (P < 0.01) with increasing level of P from either source. The main effect of P source (MSP vs. MBM) was not significant, except for the greater femur strength (P < 0.05) in the pigs fed the MSP-supplemented diets. Femur and metacarpal/metatarsal strength and metacarpals ash (grams) were regressed on grams of added P consumed for each P source, with the basal included in both regressions. Based on slope ratios (MSP considered as 100%), the relative bioavailability of P in MBM averaged 87% when the regression lines were forced through a common intercept and 95% when unforced. In Exp. 2, 100 pigs were fed fortified corn-soybean meal or corn-soybean meal-MBM diets from 45 to 110 kg BW to evaluate MBM as the sole source of supplemental P. The MBM (54% CP, 2.3% lysine, 9.2% Ca, 4.4% P; air-dry basis) was substituted for corn and soybean meal on a lysine basis, and crystalline lysine was added to all diets at 0.15%. Tryptophan was included in diets containing MBM. Treatments were arranged in a 2 x 2 factorial with P source (dicalcium phosphate or MBM) and P level as the two factors. The two levels of P and Ca were at the NRC requirement or the NRC level plus 0.10% additional P and Ca. Performance, carcass traits, and bone strength were not affected by source of P and Ca, but bone strength was greater (P < 0.01) at the higher P and Ca level. These results indicate that the bioavailability of P in MBM, relative to that in MSP, is high (approximately 91%) for growing pigs, and MBM can serve as the sole source of supplemental P and Ca for finishing pigs. PMID- 15827251 TI - An improved method for a rapid determination of phytase activity in animal feed. AB - The current direct colorimetric assay for phytase activity in feeds has interference from high P background and other factors. Our objective was to develop a rapid and reliable spin column method to accurately determine phytase activity in feed ingredients or complete diets. After the feed sample was extracted by stirring in 0.2 M citrate buffer, pH 5.5, for 30 min at room temperature, the oily layer of the supernatant fraction was removed by passing through an acrodisc syringe filter (0.45-microm HT Tuffryn membrane, Gelman Laboratory, Ann Arbor, MI). The filtrate was then loaded onto a spin column (MW cutoff 30,000, Millipore, Bedford, MA) to remove free phosphate before the phytase activity assay. Compared with the direct assay, this new procedure improved both accuracy and reproducibility. When diets contained phytase at 0 to 1,500 U/kg (as fed), the CV for multiple assays of the same samples (n = 6) by the new method ranged from 1 to 6% compared with 28 to 39% by the direct method. A linear relationship was found between the added phytase activity in practical diets and the analyzed activity by the new method (r2 = 0.99; P < 0.01). In conclusion, the spin column method is an improved assay for phytase activity in animal feed, and may be used for quality control of phytase supplementation. PMID- 15827252 TI - Glufosinate herbicide-tolerant (LibertyLink) rice vs. conventional rice in diets for growing-finishing swine. AB - Genetically modified (GM) rice (LibertyLink, event LLRICE62) that is tolerant to glufosinate ammonium (Liberty) herbicide was compared with a near-isogenic (NI) conventional medium-grain brown rice (cultivar, Bengal) and a commercially milled long-grain brown rice in diets for growing-finishing pigs. The GM and NI rice were grown in 2000. The GM rice was from fields treated (GM+) or not treated (GM ) with glufosinate herbicide. The GM- and NI rice were grown using herbicide regimens typical of southern United States rice production practices. The four rice grains were similar in composition. Growing-finishing pigs (n = 96) were fed fortified rice-soybean meal diets containing the four different rice grains from 25 to 106 kg BW. Diets contained 0.99% lysine initially (growing phase), with lysine decreased to 0.80% (early finishing phase) and 0.65% (late finishing phase), when pigs reached 51 and 77 kg, respectively. The percentage of rice in the four diets was constant during each of the three phases (72.8, 80.0, and 85.8% for the growing, early-finishing, and late-finishing phases, respectively). There were six pen replicates (three pens of barrows and three pens of gilts) and four pigs per pen for each dietary treatment. All pigs were slaughtered at the termination of the study to collect carcass data. At the end of the 98-d experiment, BW gain, feed intake (as-fed basis), and feed:gain ratio did not differ (P > 0.05) for pigs fed the GM+ vs. conventional rice diets, but growth performance traits of pigs fed the GM+ rice diets were superior (P < 0.05) to those of pigs fed the GM- rice diet (ADG = 0.86, 0.79, 0.81, and 0.85 kg/d; ADFI = 2.41, 2.49, 2.37, and 2.45 kg/d; feed:gain = 2.80, 3.17, 2.95, and 2.89 for GM+, GM-, NI, and commercially milled rice, respectively). Carcass traits (adjusted for final BW) did not differ (P = 0.10) among treatments (hot carcass yield = 73.5, 72.6, 72.6, and 73.2%; 10th-rib backfat = 23.0, 22.7, 21.3, and 23.8 mm; LM area = 38.6, 38.0, 38.2, and 38.1 cm(2); carcass fat-free lean = 50.5, 50.5, 51.2, and 50.0%). Gilts grew slower (P < 0.05) and were leaner (P < 0.05) than barrows. Responses to type of rice did not differ between barrows and gilts, with no evidence of a diet x gender interaction (P = 0.50) for any trait. The results indicate that the glufosinate herbicide-tolerant rice was similar in composition and nutritional value to conventional rice for growing-finishing pigs. PMID- 15827253 TI - Splanchnic metabolism of nitrogenous compounds and urinary nitrogen excretion in steers fed alfalfa under conditions of increased absorption of ammonia and L arginine supply across the portal-drained viscera. AB - Effects of increased ammonia and/or arginine absorption across the portal-drained viscera (PDV) on net splanchnic (PDV and liver) metabolism of nitrogenous compounds and urinary N excretion were investigated in six catheterized Hereford x Angus steers (501 +/- 1 kg BW) fed a 75% alfalfa:25% (as-fed basis) corn soybean meal diet (0.523 MJ of ME/[kg BW(0.75).d]) every 2 h without (27.0 g of N/kg of dietary DM) and with 20 g of urea/kg of dietary DM (35.7 g of N/kg of dietary DM) in a split-plot design. Net splanchnic flux measurements were obtained immediately before beginning and ending a 72-h mesenteric vein infusion of L-arginine (15 mmol/h). For 3 d before and during arginine infusion, daily urine voided was measured and analyzed for N composition. Feeding urea increased PDV absorption (P < 0.01) and hepatic removal (P < 0.01) of ammonia N, accounting for 80% of increased hepatic urea N output (P < 0.01). Numerical increases in net hepatic removal of AA N could account for the remaining portion of increased hepatic urea N output. Arginine infusion increased hepatic arginine removal (P < 0.01) and hepatic urea N output (P < 0.03) and switched hepatic ornithine flux from net uptake to net output (P < 0.01), but numerical changes in net hepatic removal of ammonia and AA N could not account fully for the increase in hepatic urea N output. Increases in urine N excretion equaled quantities of N fed as urea or infused as arginine. Estimated salivary urea N excretion was not changed by either treatment. Urea cycle regulation occurs via a complex interaction of mechanisms and requires N sources other than ammonia, but the effect of increased ammonia absorption on hepatic catabolism of individual AA in the present study was not significant. PMID- 15827254 TI - Splanchnic metabolism of nutrients and hormones in steers fed alfalfa under conditions of increased absorption of ammonia and L-arginine supply across the portal-drained viscera. AB - Effects of increased ammonia and/or arginine absorption on net splanchnic (portal drained viscera [PDV] plus liver) metabolism of nonnitrogenous nutrients and hormones in cattle were examined. Six Hereford x Angus steers (501 +/- 1 kg BW) prepared with vascular catheters for measurements of net flux across the splanchnic bed were fed a 75% alfalfa:25% (as-fed basis) corn and soybean meal diet (0.523 MJ of ME/[kg BW(0.75).d]) every 2 h without (27.0 g of N/kg of DM) and with 20 g of urea/kg of DM (35.7 g of N/kg of DM) in a split-plot design. Net flux measurements were made immediately before and after a 72-h mesenteric vein infusion of L-arginine (15 mmol/h). There were no treatment effects on PDV or hepatic O2 consumption. Dietary urea had no effect on splanchnic metabolism of glucose or L-lactate, but arginine infusion decreased net hepatic removal of L lactate when urea was fed (P < 0.01). Net PDV appearance of n-butyrate was increased by arginine infusion (P < 0.07), and both dietary urea (P < 0.09) and arginine infusion (P < 0.05) increased net hepatic removal of n-butyrate. Dietary urea also increased total splanchnic acetate output (P < 0.06), tended to increase arterial glucagon concentration (P < 0.11), and decreased arterial ST concentration (P < 0.03). Arginine infusion increased arterial concentration (P < 0.07) and net PDV release (P < 0.10) and tended to increase hepatic removal (P < 0.11) of insulin, as well as arterial concentration (P < 0.01) and total splanchnic output (P < 0.01) of glucagon. Despite changes in splanchnic N metabolism, increased ammonia and arginine absorption had little measurable effect on splanchnic metabolism of glucose and other nonnitrogenous components of splanchnic energy metabolism. PMID- 15827256 TI - Non-steady-state modeling of effects of timing and level of concentrate supplementation on ruminal pH and forage intake in high-producing, grazing ewes. AB - A computer model was developed to predict responses of lactating ewes to concentrate supplementation, whether on pasture or stall-fed, given concentrate once per day or in multiple feedings, and suckling multiple lambs. The model considers effects of concentrate supplementation on organic acid production, saliva flow, ruminal pH, and forage intake. The user defines ewe BW, feed composition, and concentrate feeding times and amounts. The reference ewe has free access to forage and water. Upon consumption, forages and concentrates enter into lag pools for 2.0 and 0.24 h, respectively. Carbohydrates then enter ruminal pools of degradable fiber, undegradable fiber, or nonstructural carbohydrate, from which they are degraded or pass to the lower gut. Rapid dissociation of organic acids from carbohydrate fermentation and buffers from rumination are simulated to determine ruminal pH according to the Henderson-Hasselbach equation. The pH, in turn, affects fiber degradation rates. Forage intake continues during daylight hours until ruminal NDF exceeds 1.0% of BW, or organic acid concentration exceeds 130 mM. A circadian pattern of organic acid concentrations and pH of rumen contents with multiple concentrate feedings was simulated by the model with root mean square prediction error of 7.7 and 3.0 to 4.0% of the observed mean, respectively. However, ignoring fermentation of dietary protein may have caused an underestimation of organic acid production rates. The model predicted the increase in total DMI and the substitution effect on forage intake of increasing levels of concentrate supplementation. Simulations suggested that a single concentrate meal daily was best fed in the evening to minimize the substitution effect, and that there was no benefit in forage intake to feeding 2 kg/d concentrate in more than two meals per day. PMID- 15827255 TI - Influence of fish oil supplementation on growth and immune system characteristics of cattle. AB - A study was conducted to determine the effects of supplemental fish oil on growth performance and immune system characteristics of beef calves. The grazing phase (78 d) used 48 yearling crossbred steers (231 +/- 22 kg initial BW) grazing 0.45 ha mixed-grass pastures (four per treatment) supplemented with 1.82 kg/d (as-fed basis) of the diets. Diets consisted of 1) corn-based supplement; 2) corn-based supplement with 1.5% (as-fed basis) fish oil; 3) wheat midd-based supplement; and 4) wheat midd-based supplement with 1.5% fish oil. On d 78, all calves were bled by jugular venipuncture, and blastogenic response of peripheral blood lymphocytes to phytohemagglutinin, concanavalin A, and pokeweed mitogen was measured. Fish oil supplementation negatively affected ADG with the corn-based supplement, but it had no effect when added to the wheat midd-based supplement (base-supplement x fish oil interaction; P < 0.03). Isolated lymphocytes from calves fed the corn based supplement with fish oil had a greater response to stimulation with concanavalin A than did lymphocytes from calves fed the corn-based supplement alone, but there was no effect of fish oil addition to the wheat midd-based supplement (base-supplement x fish oil interaction; P < 0.01). During the growing phase, the 48 steers (352 +/- 32 kg initial BW) from the grazing phase were moved to drylot pens and were stratified by BW and previous dietary treatment (three calves per pen; eight pens per dietary treatment) for a 56-d growing trial. Dietary treatments consisted of 1) control, and 2) the control diet with 3% (as fed basis) fish oil. Calves supplemented with fish oil had decreased ADG, ADFI, and G:F (P < or = 0.02) compared with controls. Fish oil supplementation during the grazing phase modulated the immune system; however, the decreased growth performance associated with fish oil in both trials may limit its practical use as an immune stimulant. PMID- 15827257 TI - Effect of rapid or gradual grain adaptation on subacute acidosis and feed intake by feedlot cattle. AB - The effects of grain adaptation protocol on subacute acidosis and feed intake by cattle were studied in a completely randomized experiment using 12 crossbred heifers (384 +/- 25 kg BW). The dietary proportion of concentrate was increased from 40 to 90% (DM basis) either by rapid adaptation (65% concentrate diet fed for 3 d) or by gradual adaptation (five intermediate diets containing 48.3, 56.7, 65.0, 73.3, and 81.7% concentrate, fed for 3 d each). Feed intake and ruminal pH (by indwelling ruminal electrodes) were monitored over 20 d. Mean daily pH variables did not differ (P > or = 0.10) between treatments on any of the 3 or 4 d that 65 or 90% concentrate was fed. Variances of a number of pH variables were greater (P < 0.05) for rapidly adapted heifers than for those on the gradual adaptation protocol during adaptation to 65 and 90% concentrate. Mean hourly pH did not differ over the first 24 h of adaptation to 65% concentrate, but variance of hourly pH tended (P < 0.10) to be greater for rapidly adapted than for gradually adapted heifers for eight of the first 24 h. On the first day of feeding 90% concentrate, ruminal pH tended (P = 0.07) to be less at 11 and 12 h after feeding with rapid adaptation than with gradual adaptation. Variance of hourly pH increased steadily in rapidly adapted heifers from 6 h after feeding onward. Ruminal VFA concentration and osmolality did not differ between treatments. Ruminal lactate concentration was < 1 mM, except in two rapidly adapted heifers and one gradually adapted heifer after introduction to 90% concentrate. Adaptation method did not affect DMI or day-to-day variation in DMI. Detection of acidosis was associated with increased variance in ruminal pH variables. A range of individual responses to grain challenge was observed, but current management strategies for preventing acidosis in pens of cattle are based on responses of the most susceptible individuals. A better understanding of factors governing individual responses to acidotic challenge may allow for the development of more effective acidosis prevention practices. PMID- 15827258 TI - Improving the nutritional value of oat hulls for ruminant animals with pretreatment of a multienzyme cocktail: in vitro studies. AB - Relatively high amounts of hydroxycinnamic acid in oat hulls, mainly ferulic acid, are believed to be inhibitory to digestion by ruminal microorganisms. Ferulic acid is produced via the phenylpropanoid biosynthetic pathway and covalently cross-linked to polysaccharides by ester bonds and to components of lignin, mainly by ether bonds. Ferulic acid also forms dimers or trimers. As a result, polysaccharides become extensively cross-linked by ferulate dimerization or trimerization and incorporation into lignin. Previous studies have shown that Aspergillus ferulic acid esterase and Trichoderma xylanase act synergistically to release ferulic acid from feruloyl-polysaccharides in complex plant cell walls of oat hulls. This activity opens the remainder of the polysaccharides to further hydrolytic attack and facilitates the accessibility of the main polysaccharide chain to cellulase, thereby increasing the release of reducing sugars. In Exp. 1, the best multienzyme cocktail (ferulic acid esterase, xylanase, cellulase, endo glucanase [I, II], and beta-glucanase) was developed using an orthogonal experimental design, L25 (5(6)), where L = orthogonal table; 6 = factors; 5 = five levels of each; and 25 = experimental number, for further in situ and/or in vivo study. In Exp. 2, in vitro biodegradation studies with a 3 x 2 x 4 factorial arrangement of treatments were used to evaluate the responses of three feedstuffs, oat hulls or standard references (wheat straw and alfalfa hay), two particle sizes (1 mm and 250 microm), and four in vitro incubation treatments with the best multienzyme cocktail developed in Exp. 1. Addition of the multienzyme cocktail to the forages improved (P < 0.01) in vitro ruminal fluid degradability. With respect to feedstuff, the order of response (P < 0.05) to the treatments was oat hulls (+12% unit) > wheat straw (+5% unit) > alfalfa (+2% unit). This multi-enzyme cocktail seems best suited for oat hulls containing feruloyl ester bonds. In conclusion, data from this study suggest that the addition of the multienzyme cocktail to poorly digestible feeds before feeding enhanced degradation of DM. PMID- 15827259 TI - Ruminal in situ disappearance kinetics of dry matter and fiber in growing steers for common crabgrass forages sampled on seven dates in northern Arkansas. AB - Southern crabgrass (Digitaria ciliaris [Retz.] Koel.) is often viewed as an undesirable weed, largely because it encroaches upon field and forage crops, gardens, and lawns. However, visual observations of livestock grazing mixed species pastures suggest that cattle seem to prefer crabgrass to many other summer forages. The objectives of this study were to assess the nutritive value of crabgrass sampled weekly between July 11, and August 22, 2001, and then to determine ruminal in situ disappearance kinetics of DM and NDF for these crabgrass forages. A secondary objective was to compare these kinetic estimates with those of alfalfa (Medicago sativa L.), bermudagrass (Cynodon dactylon [L.] Pers.), and orchardgrass (Dactylis glomerata L.) control hays. All forages were evaluated in situ using five (383 +/- 22.7 kg) ruminally cannulated crossbred (Gelbvieh x Angus x Brangus) steers. Whole-plant crabgrass exhibited more rapid (P < or = 0.002) ruminal disappearance rates of DM (overall range = 0.069 to 0.084 h(-1)) than did bermudagrass (0.054 h(-1)) and orchardgrass (0.060 h(-1)) hays, but disappearance rates were slower (P < 0.001) for crabgrass than for alfalfa hay (0.143 h(-1)). Effective ruminal disappearance of DM was greater (P < 0.001) for crabgrass (overall range = 69.3 to 75.4%) than for all the control hays. Similarly, disappearance rates of NDF for crabgrass (overall range = 0.069 to 0.086 h(-1)) were more rapid (P < 0.001) than observed for bermudagrass and orchardgrass hays; however, NDF in alfalfa disappeared at a faster (P < 0.001) rate (0.107 h(-1)) than crabgrass. These results indicate that crabgrass offers greater effective ruminal degradability of DM and NDF than orchardgrass or alfalfa of moderate quality. More importantly, it potentially offers faster and more extensive ruminal disappearance than perennial warm-season grasses typically found throughout the southeastern United States, and it should likely support improved performance by ruminant livestock in this region. PMID- 15827260 TI - Fatty acid indices of stearoyl-CoA desaturase do not reflect actual stearoyl-CoA desaturase enzyme activities in adipose tissues of beef steers finished with corn , flaxseed-, or sorghum-based diets. AB - We hypothesized that stearoyl-CoA desaturase (SCD) enzyme activity would not correlate with fatty acid indices of SCD activity in steers fed different grains. Forty-five Angus steers (358 +/- 26 kg BW) were individually fed for 107 d diets differing in whole cottonseed (WCS) supplementation (0, 5, or 15% of DM) and grain source (rolled corn, flaxseed plus rolled corn, or ground sorghum grain) in a 3 x 3 factorial arrangement. Flaxseed- and corn-fed steers had greater (P < 0.01) G:F (0.119 and 0.108, respectively) than sorghum-fed steers (0.093). Marbling score was decreased by WCS (P = 0.04), and LM area was decreased (P < 0.01) by sorghum. Plasma 14:0, 16:0, 16:1n-7, and 18:2n-6 were greatest in corn fed steers, whereas plasma 18:3n-3 and 20:5n-3 were greatest in the flax-seed-fed steers (P < 0.01). Plasma 18:1trans-11 was least in sorghum-fed steers, and plasma cis-9,trans-11 CLA was barely detectable, in spite of high intestinal mucosal SCD enzyme activity (118 to 141 nmol*g tissue(-1).7 min(-1)). Interfascicular (i.f.) and s.c. cis-9,trans-11 CLA remained unchanged (P > or = 0.25) by treatment, although 18:1trans-11 was increased (P < or = 0.02) in steers fed corn or flaxseed. Steers fed flaxseed also had greater (P < 0.01) i.f. and s.c. concentrations of 18:3n-3 than steers fed the other grain sources. Oleic acid (18:1n-9) was least and total SFA were greatest (P < 0.01) in i.f. adipose tissue of steers fed 15% WCS. Lipogenesis from acetate in s.c. adipose tissue was greater (P < 0.01) in flaxseed-fed steers than in the corn- or sorghum-fed steers. Steers fed flaxseed or corn had larger i.f. mean adipocyte volumes (P < 0.01) than those fed sorghum and tended (P = 0.07) to have larger s.c. adipocyte volumes. Several fatty acid indices of SCD enzyme activity were decreased (P < or = 0.03) by WCS in i.f. adipose tissue, including the 18:2cis-9,trans-11/ 18:1trans-11 ratio. The 18:2cis-9,trans-11/18:1trans-11 ratio also tended to be decreased (P = 0.09) in s.c. adipose tissue by flaxseed; however, SCD enzyme activities in i.f. and s.c. adipose tissue were not affected by dietary WCS (P > or = 0.47) or grain source (P > or = 0.37). Differences in SFA seemed to be independent of SCD enzyme activity in both adipose tissues, suggesting that duodenal concentrations of fatty acids were more important in determining tissue fatty acid concentrations than endogenous desaturation by SCD. PMID- 15827261 TI - The fatty acid composition of muscle fat and subcutaneous adipose tissue of pasture-fed beef heifers: influence of the duration of grazing. AB - Our objective was to determine the effect of the duration of grazing before slaughter on the fatty acid composition of muscle fat and s.c. adipose tissue (SAT) of beef heifers. Sixty crossbred Charolais heifers (n = 15 per treatment) were assigned randomly to one of four dietary treatments: 45 animals (Treatments 1, 2, and 3, respectively) were housed at the beginning of the experiment, and 15 (Treatment 4) were fed at pasture. Two groups of 15 heifers were moved to pasture 40 d (Treatment 2) and 99 d (Treatment 3) before slaughter, respectively, resulting in preslaughter grazing periods of 0, 40, 99, or 158 d for Treatments 1, 2, 3, and 4, respectively. Before grazing the predominantly perennial ryegrass pasture, animals were housed and offered grass silage ad libitum and 3 kg of concentrate diet (650 g of grass silage/kg of total DMI). After slaughter, the fatty acid profile of the neutral (NL) and polar lipid (PL) fractions of muscle fat from the LM and the total lipids from SAT were analyzed by gas chromatography. Duration of grazing showed a quadratic tendency on mean carcass weight (P = 0.08), but did not affect growth (P = 0.27) or the lipid content (P = 0.13) of the LM. Increasing the duration of grazing led to a linear increase (P < 0.001) in the concentration (on fresh-tissue basis) of CLA in muscle fat (from 11.80 to 17.75 mg/100 g of muscle in NL, and from 0.52 to 0.82 mg/100 of g muscle in PL) and in SAT (from 3.98 to 10.23 mg/g of SAT; P < 0.001), and increased the concentration of C18:1trans-11 in both muscle fat fractions (P < 0.001) and in SAT (P < 0.001). In the total muscle lipids, the polyunsaturated to saturated fatty acid ratio (P:S) increased from 0.12 to 0.15 with increased duration of grazing following a linear (P < 0.05) and cubic pattern (P < 0.05). Increasing the duration of grazing led to a linear decrease in the n-6:n-3 ratio of muscle fat from 2.00 to 1.32 (P < 0.001), and from 2.64 to 1.65 in the SAT lipids (P < 0.001), mainly as a consequence of the increased concentration of C18:3n-3. It is concluded that muscle fat and SAT fatty acid profile was improved from a human health perspective by pasture feeding, and that this improvement depended on the duration of grazing. PMID- 15827262 TI - Intake, digestion, and N metabolism in steers fed endophyte-free, ergot alkaloid producing endophyte-infected, or nonergot alkaloid-producing endophyte-infected fescue hay. AB - A digestion and N balance trial was conducted to compare effects of traditional endophyte-infected (E+), endophyte-free (E-), and nontoxic endophyte infected (NE; MaxQ; Pennington Seed, Inc., Madison, GA) Jesup tall fescue (Festuca arundinacea Schreb.) hay on digestion and N retention in steers. Hay composition (DM basis) was as follows: E+ (10.8% CP, 59.9% NDF, and 29.4% ADF), E- (11.8% CP, 58.5% NDF, and 28.4% ADF), and NE (11.6% CP, 58.6% NDF, and 28.3% ADF). Eight Polled Hereford steers (initial BW 240 +/- 9 kg) were used in a replicated, 3 x 3 Latin square design, with an extra steer allotted to each square. Steers were fed ad libitum for 14 d, followed by a 9-d adaptation to restricted intake (based on the animal with the lowest ad libitum intake for the square) and a 5-d fecal and urine collection. Water intake (20.2 L/d) and urine output (7.40 L/d) did not differ (P > 0.10) during the collection period. Plasma prolactin concentration was less (P < 0.05) for steers on the E+ hay (8.83 ng/mL) than for those on the E hay (18.03 ng/mL) and intermediate for steers on the NE hay (12.65 ng/mL). Endophyte-infected hay differed (P < 0.05) from E- and NE in ad libitum DMI (5.02 vs. 5.62 and 5.61 kg/d, respectively) and ad libitum DMI as a percentage of BW (1.86 vs. 2.06 and 2.06%, respectively). Restricted DMI during the fecal and urine collection was lower (P < 0.05) for E+ hay than for E- (5.04 vs. 5.24 kg/d), and NE was intermediate (5.19 kg/d). Dry matter digestibility was lower (P < 0.05) for E+ compared with E- and NE (62.3 vs. 67.0 and 65.9%, respectively). Digestibility of ADF was lower (P < 0.05) for E+ than for E-, and was intermediate for NE (61.5, 66.0, and 63.9%, respectively). There were no differences for NDF, cellulose, or hemicellulose digestibilities among hay types. Crude protein digestibility was higher (P < 0.05) for E- and NE than for E+ (54.3 and 52.5 vs. 48.1%, respectively). Nitrogen retention was lower (P < 0.01) for E+ than for E- or NE (15.6 vs. 22.7 or 23.0 g/d, respectively). Hay type did not influence plasma urea N, urine urea N output, or urine urea N as a percentage of urinary N. Results from this study indicate that E+ tall fescue hay was lower in ad libitum DMI, DM digestibility, and N retention than NE or E- hays with similar chemical composition. Hay from NE and E- fescue had nearly identical composition, and did not differ for any variable measured. PMID- 15827263 TI - Time-dependent influence of supranutritional organically bound selenium on selenium accumulation in growing wether lambs. AB - Crossbred wethers (n = 36; BW = 36.0 kg; SD = 3.4) were used to assess the time dependent influence of supranutritional organically bound Se on Se accumulation. Four wethers were slaughtered before the trial began (d 0). The remaining wethers were fed diets containing adequate (0.2 microg of Se/g of DM) or supranutritional Se (2.9 microg of Se/g of DM; in the form of high-Se wheat grain) for 14, 28, 42, or 56 d before slaughter (four wethers per Se treatment at each slaughter day). The DMI was set at 3.1% of BW and adjusted weekly based on a targeted ADG of 150 g. Daily Se intake by wethers fed the adequate and supra-nutritional Se diets ranged from 5.3 to 5.9, and 79.0 to 95.0 microg of Se/kg of BW, respectively, and did not differ (P = 0.84 to 0.99) between slaughter day groups within Se treatment. Neither Se treatment nor Se treatment x slaughter day interactions were significant for BW, G:F, or liver, kidneys, and spleen weights (P = 0.06 to 0.84). Within the supranutritional Se treatment, Se contents of most organs and tissues from wethers slaughtered on d 14, 28, 42, and 56 were nearly twice the concentrations (P < 0.01) of wethers slaughtered on d 0. When regressed against the number of days the wethers were fed supranutritional Se, Se concentrations increased (P < 0.001) cubically in kidneys and plasma, quadratically in duodenum, lung, liver, and spleen, and linearly in heart, muscle, and wool. For total Se in kidneys, liver, and spleen, the response was quadratic (P < 0.03). Excluding skeletal muscle, heart, and wool, Se in other organs and tissues reached apparent steady-state concentrations 14 to 28 d after commencement of supranutritional Se diets. Selenium concentrations in skeletal muscle accumulated in a linear manner (P < 0.001) throughout the 56-d feeding period. High-Se grains can be used strategically to deliver supranutritional Se and rapidly enhance Se depots in sheep, a task that does not seem attainable with Se salts. Furthermore, a 100-g portion of uncooked loin (LM) from the wethers fed supranutritional Se contained 196 to 250% of the recommended Se requirement for humans. PMID- 15827264 TI - Relationship of sire expected progeny differences to milk yield in Brangus cows. AB - Milk yield from 160 Brangus cows sired by 65 Brangus bulls was measured over a 3 yr period with a single-cow milking machine to estimate the relationship of actual milk yield of daughters and their calves' BW with cow sire EPD for milk during the preweaning period. Milk yield was measured six times per year at an average 49, 78, 109, 138, 168, and 198 d postpartum. The regression of daughters' milk yield on sire milk EPD was quadratic (P < 0.01), and the initial linear portion of the curve differed among months (P < 0.05) at an average cow BW. Similarly, the regression of 6-mo average 24-h milk yield on sire milk EPD was curvilinear (P < 0.05). When cow BW was fitted as a covariate in the regression of 6-mo average 24-h milk yield on sire milk EPD, there was an interaction of cow BW with linear sire milk EPD and quadratic sire milk EPD (P < 0.10). The associated response surface suggested that the regression was primarily linear in cows weighing < or = 520 kg and curvilinear in cows weighing >520 kg. A trend existed for the regression of calf 205-d weight on grandsire milk EPD to be curvilinear (P < 0.21); however, the regression of calf 205-d weight on milk yield of their dam was linear (P < 0.01). Results from these data suggest that genetic potential for milk yield, and possibly the associated effects on calf BW transmitted through the grandsire, may have a practical maximum because of nutritional limitations that prevent the expression of genetic potential beyond that level, particularly in heavier cows, which suggests the need to match sire milk EPD and cow BW with production environment. PMID- 15827265 TI - Duration of restraint and isolation stress as a model to study the dark-cutting condition in cattle. AB - Holstein steer calves (n = 32; 156 +/- 33.2 kg average BW) were used to evaluate the duration of restraint and isolation stress (RIS) on endocrine and blood metabolite status and the incidence of dark-cutting LM. Calves were blocked by BW and assigned randomly within blocks to one of four stressor treatments: unstressed controls (NS) or a single bout of RIS for 2, 4, or 6 h. Venous blood was collected via indwelling jugular catheters at 40, 20, and 0 min before stressor application and at 20-min intervals during RIS. Unstressed calves remained in their home stanchions and, except for blood sampling, were subjected to minimal handling and stress. Serum cortisol and plasma lactate concentrations were increased (P < 0.01) during the first 20 min after RIS application, and remained elevated throughout the 6 h of RIS. Plasma concentrations of glucose and insulin were greater (P < 0.05) in RIS calves than in NS calves after 80 and 100 min of stressor application, respectively; however, RIS did not (P > 0.80) affect plasma NEFA concentrations. Calves were slaughtered within 20 min of completion of RIS, and muscle samples were excised from right-side LM at 0, 0.75, 1.5, 3, 6, 12, 24, and 48 h after exsanguination for quantifying LM pH, and glycogen and lactate concentrations. The pH of the LM from calves subjected to 6 h of RIS exceeded 6.0, and was greater (P < 0.05) at 24 and 48 h postmortem than the pH of NS calves or calves subjected to 2 or 4 h RIS. Muscle glycogen concentrations did not differ (P = 0.16; 25.58, 10.41, 13.80, and 14.41 micromol/g of wet tissue weight for NS and 2-, 4-, and 6-h RIS, respectively), and LM lactate concentrations tended to be lower (P = 0.08) in calves subjected to 6 h of RIS. At 48 h after exsanguination, the LM from calves subjected to 6 h of RIS had more (P < 0.05) bound and less (P < 0.05) free moisture than did the LM from NS calves or calves subjected to 2 or 4 h of RIS. Additionally, the LM from RIS calves was darker (lower L* values; P < 0.05) than the LM of NS calves. Visual color scores for the LM were greatest (P < 0.05) for calves subjected to 6 h of RIS and least (P < 0.05) for NS calves. Subjecting lightweight Holstein calves to 6, 4, and 2 h of RIS resulted in six (75%), two (25%), and two (25%) carcasses characteristic of the dark-cutting condition, respectively. There were no dark-cutting carcasses produced from NS calves. Thus, RIS may be a reliable animal model with which to study the formation of the dark-cutting condition. PMID- 15827266 TI - Cost evaluation of the use of conventional and electronic identification and registration systems for the national sheep and goat populations in Spain. AB - A cost model was developed to compare different implementation strategies of the new European Commission regulation for sheep and goat identification and registration (EC 21/2004) in Spain. Strategies were as follows: 1) conventional identification (CID) by two ear tags; 2) electronic identification (EID) by one bolus and one ear tag; and 3) mixed CID and EID strategy (MID), consisting of CID for fattening stock and EID for breeding stock. Complete and simplified implementations of the regulation were considered as options. Total costs per animal identified for all strategies and options varied according to the implementation option, ranging from Euros 2.48 and 4.64. The EID was the most expensive strategy (Euros 4.47 to 4.64) for all implementation options. Cost of CID and MID strategies ranged from Euros 2.63 to 2.98 and from Euros 2.48 to 3.03, respectively. The model was submitted to a sensitivity analysis without considering extra benefits of sheep and goat identification. Critical values for which the cost of MID equaled CID depended on strategy and option, and ranged from 7.5 to 11.5% for ear tag losses and from Euros 1.80 to 3.30 for bolus price. In conclusion, the use of a mixed strategy combining conventional ear tags (animals intended for slaughter) and electronic boluses (breeding stock) seems to be an affordable strategy that fulfills the European Commission regulation requirements for the identification of sheep and goats in Spain. Price reductions for devices and equipment would make the full electronic identification strategy less expensive in the future. PMID- 15827267 TI - Increased levels and reduced catabolism of asymmetric and symmetric dimethylarginines in pulmonary hypertension. AB - Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase (NOS) and has been implicated in endothelial dysfunction. ADMA is metabolized by the enzyme dimethylarginine dimethylaminohydrolase (DDAH), with DDAH2 representing the predominant endothelial DDAH isoform. Symmetric dimethylarginine (SDMA), also originating from arginine methylation by protein arginine methyltransferases, is an inhibitor of intracellular arginine uptake. In both chronic pulmonary hypertensive rats and patients suffering from idiopathic pulmonary arterial hypertension (IPAH; NYHA class III and IV), a marked increase in plasma ADMA and SDMA levels, as well as tissue levels of asymmetric and symmetric dimethylated proteins, was observed. Moreover, when comparing lung tissue from pulmonary hypertensive rats and IPAH patients to corresponding normal lung tissue, expression of DDAH2 was found to be reduced at both the mRNA and the protein level with no significant changes in DDAH1 expression. These findings were further supported by demonstrating a decrease in DDAH2 function in the experimental pulmonary hypertension model. Immunohistochemistry in human and rat control tissue demonstrated both isoforms of DDAH in the endothelial layer and in the alveolar epithelium. In contrast, in pulmonary hypertensive tissue, the immunoreactivity of DDAH2 in pulmonary endothelium was significantly decreased compared with DDAH1. Therefore, altogether we can conclude that enhanced dimethylarginine levels may contribute to vascular abnormalities in pulmonary arterial hypertension. Suppression of endothelial DDAH2 expression and function represents an important underlying mechanism. PMID- 15827268 TI - Toxicological research involving humans: ethical and regulatory considerations. PMID- 15827269 TI - Spider bites: addressing mythology and poor evidence. PMID- 15827271 TI - Hypercalcemia in hepatic tuberculosis: a case report in Korea. AB - Although primary hyperparathyroidism and malignant diseases account for approximately 90% of the causes of hypercalcemia, they could occur in association with granulomatous diseases such as tuberculosis or sarcoidosis, especially in developing countries. Hepatic tuberculosis is difficult to diagnosis without suspicion in cases with normal findings on chest radiographs. We report a 70-year old woman who presented with hypercalcemia due to hepatic tuberculosis. The diagnosis was made by a computed tomography scan and laparoscopic evaluation. After treatment with anti-tuberculosis medication, her hypercalcemia resolved. Increased vitamin D synthesis by activated macrophages in the granuloma tissue is the major mechanism of hypercalcemia in tuberculosis. PMID- 15827272 TI - Association between sex, nutritional status, severity of dengue hemorrhagic fever, and immune status in infants with dengue hemorrhagic fever. AB - The association between sex, nutritional status, and the severity of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS), and immune status was investigated in 245 Vietnamese infants with predominantly primary infections with dengue virus. Male and female infants were at equal risk of developing DHF/DSS. However, infants of low height and weight for age were under-represented among DHF/DSS cases compared with 533 healthy baby clinic infant controls. Acute illness phase blood levels of selected cytokines (interferon-gamma and tumor necrosis factor-alpha) and serum levels of antibodies to dengue virus were elevated in the same range in male and female infants with DHF/DSS, as well as in infants with and without malnutrition. PMID- 15827273 TI - Characterization of dengue-2 virus binding to surfaces of mammalian and insect cells. AB - The binding of dengue type 2 (DEN-2) virus to mammalian (LLC-MK(2) and Vero) and mosquito (C6/36 and AP61) cell surfaces was investigated by a virus-binding assay using purified (3)H-labeled DEN-2 virus. The DEN-2 virus binding to all four cell types was specific and saturable, indicating the presence of a single class of receptors (ranging from 3.7 x 10(3) to 3.5 x 10(4) receptors/cell) with a high affinity for DEN-2 virus (K(d) ranging from 98 to 171 pM). Treatment of cell surfaces with certain glycosidases significantly reduced virus binding to mammalian cell lines, but not to the insect cell lines examined. Furthermore, heparin was found to compete with mammalian cell receptors for binding to DEN-2 virus and to inhibit viral infection of mammalian cells, but heparin had no effect on viral binding to or infection of insect cells. These results confirm previous reports suggesting that DEN-2 virus receptors on mammalian cell lines are different from those on insect cell lines. PMID- 15827274 TI - Ecologic covariates of hookworm infection and reinfection in rural Kwazulu natal/south Africa: a geographic information system-based study. AB - Information on hookworm infection and re-infection in a cohort of primary school children and interview data on their socioeconomic background and behavior were combined with environmental data using a geographic information system (GIS). Multivariate models served to explore the covariation of environmental and infection patterns adjusted for possible confounders. Our aim was to identify environmental factors that might serve to predict infection and thus guide control efforts when epidemiologic information is insufficient. Furthermore, we wanted to establish whether soil type has a genuine influence on hookworm infection. Prevalence maps and spatial statistics showed considerable spatial clustering of infection in the small ( approximately 28 x 16 km) study area. The multivariate logistic regression models showed strong positive associations of infection at baseline (baseline prevalence = 83.2%) with settlement density (odds ratio [OR] = 1.24, 95% confidence interval [CI] = 1.10-1.38) and vegetation density (OR = 1.66, 95% CI = 1.25-2.22) and a strong negative association with the clay content of the soil (OR = 0.67, 95% CI = 0.62-0.73). Similar but weaker correlations were found after re-infection. Socioeconomic status and behavior did not seem to confound these associations. Spatial analysis of the model residuals suggested that because the models accounted for most of the spatial pattern, the model standard errors were not affected by spatial clustering. Our study shows that the pattern of hookworm infection is strongly influenced by several environmental factors. The GIS-aided prediction of areas in need of treatment is therefore a promising tool to guide control efforts when epidemiologic information is insufficient. PMID- 15827275 TI - Effect of irrigation and large dams on the burden of malaria on a global and regional scale. AB - Human-made ecologic transformations have occurred at an unprecedented rate over the past 50 years. Prominent among them are water resource development projects. An estimated 40,000 large dams and 800,000 small dams have been built, and 272 million hectares of land are currently under irrigation worldwide. The establishment and operation of water projects has had a history of facilitating a change in the frequency and transmission dynamics of malaria, but analyses of these environmental risk factors are sparse. Here, we present a comprehensive review of studies that assessed the impact of irrigation and dam building on malaria prevalence or incidence, stratified by the World Health Organization's (WHO) sub-regions of the world, and link these studies with the latest statistics on disability adjusted life years, irrigated agriculture, and large dams. We also present estimates of the population at risk due to proximity to irrigation schemes and large dam reservoirs. In WHO sub-regions 1 and 2, which have 87.9% of the current global malaria burden, only 9.4 million people are estimated to live near large dams and irrigation schemes. In contrast, the remaining sub-regions concentrate an estimated 15.3 million people near large dams and up to 845 million near irrigation sites, while here only 12.1% of the global malaria burden is concentrated. Whether an individual water project triggers an increase in malaria transmission depends on the contextual determinants of malaria, including the epidemiologic setting, socioeconomic factors, vector management, and health seeking behavior. We conclude that in unstable malaria endemic areas, integrated malaria control measures, coupled with sound water management, are mandatory to mitigate the current burden of malaria in locations near irrigation or dam sites. PMID- 15827276 TI - Emergence of atovaquone-proguanil resistance during treatment of Plasmodium falciparum malaria acquired by a non-immune north American traveller to west Africa. AB - The importation of drug-resistant malaria is a growing public health problem in non-endemic countries. The combination of atovaquone and proguanil (Malarone) has become established as an agent of choice to prevent and treat chloroquine resistant Plasmodium falciparum malaria in travelers. We describe the first reported case in North America of genetically confirmed atovaquone/proguanil resistant P. falciparum malaria. Polymerase chain reaction and sequence analysis of the primary and recrudescent isolates confirmed the acquisition of a point mutation (Tyr268Ser) in the cytochrome b gene of the recrudescent isolate known to confer high-level resistance to atovaquone. Suboptimal therapy may have played a contributory role in the emergence of resistance. PMID- 15827278 TI - Evaluation of 7SL RNA gene sequences for the identification of Leishmania spp. AB - We evaluated the use of 7SL RNA gene sequences for the identification of Leishmania spp. A fragment (approximately 137 basepairs) of the 7SL RNA gene from 13 reference strains and 18 clinical isolates of 11 different Leishmania species was amplified and sequenced using conserved primers. Reference strains from each Leishmania spp. complex showed unique sequences. The nucleotide sequences were compared pairwise and a range of 81.0-99.3% intercomplex similarity was observed. Clinical isolates of the same species had sequences identical to the corresponding reference strains; thus, the intraspecies similarity was 100%. A phylogenetic tree derived from the 7SL RNA gene partial sequences was constructed and is in agreement with accepted phylogenetic schemes. PMID- 15827277 TI - Decreased prevalence of the Plasmodium falciparum chloroquine resistance transporter 76T marker associated with cessation of chloroquine use against P. falciparum malaria in Hainan, People's Republic of China. AB - The use of chloroquine treatment for Plasmodium falciparum malaria was abandoned in China in 1979 because of widespread drug resistance. Subsequent studies found decreases in the prevalence of chloroquine-resistant strains. To evaluate these decreases and assess the current status of chloroquine sensitivity in Hainan, China, we determined the prevalence of the P. falciparum chloroquine resistance transporter (PfCRT) 76T marker in the DNA of blood samples collected from 1978 to 2001. Results showed the presence of PfCRT 76T in 101 of 112 samples (90%) from 1978 to 1981, 30 of 43 samples (70%) from 1986, 22 of 34 samples (65%) from 1997 to 1998, and 37 of 68 samples (54%) from 2001. The prevalence of PfCRT 76T thus progressively decreased after chloroquine was discontinued as a treatment for P. falciparum malaria (chi(2) = 5.2, P < 0.022 [1978-1981 versus 1986]; chi(2) = 7.4, P < 0.006 [1978-1981 versus 1997-1998]; and chi(2) = 28.8, P < 0.0001 [1978 1981 versus 2001]). Reduced prevalence of the PfCRT 76T marker is consistent with greater rates of chloroquine sensitivity from in vitro drug assays of blood samples in 1997 and 2001. Monitoring for continued decreases will provide valuable information for future drug-use policies in China. PMID- 15827279 TI - Short report: efficacy of pentavalent antimony for treatment of colombian cutaneous leishmaniasis. AB - Two hundred twenty-six consecutive adult patients with cutaneous leishmaniasis in Colombia were treated with Glucantime and 81% were cured. The 19% who failed treatment had received a statistically significant lower total amount of antimony (354 mg/kg) than those who were cured (405 mg/kg) (P = 0.002). Thirty-nine of the failures were re-treated with Glucantime and 59% were cured. This large experience on initial and re-treatment cure rates probably represents the values found in clinical practice. In this series, parasite resistance may have been induced by the relatively low initial dose of antimony administered to some patients. PMID- 15827280 TI - Amplification of human DNA by primers targeted to Leishmania kinetoplast DNA and post-genome considerations in the detection of parasites by a polymerase chain reaction. AB - We evaluated the Leishmania Viannia-specific primers B1-B2 to detect Leishmania in normal skin and peripheral blood monocytes of patients with active cutaneous leishmaniasis. Southern blotting and sequencing of polymerase chain reaction (PCR) products confirmed the specificity of kinetoplast DNA (kDNA) amplification from tissue fluid from healthy skin, whereas the PCR with monocytes also amplified a human sequence of a size similar (718 basepairs) to the expected kDNA product (750 basepairs), resulting in false-positive results. Although B1 was not homologous to any human DNA sequence, B2 showed homology to a human chromosome 2 intergenic region (AC010878) at positions 35,881-36,599, which are spaced 718 nucleotides apart. Amplification of the human artifact from monocyte DNA was confirmed using the primer B2 alone. Examination of other primers reported for the PCR of kDNA from various species of Leishmania showed that six of seven were homologous to human DNA sequences. These findings underscore the importance of exploiting sequencing, bioinformatics, and DNA probes to refine molecular amplification techniques and to validate the performance of primers when used for new applications. PMID- 15827281 TI - Peripheral health workers' knowledge and practices related to filarial lymphedema care: a study in an endemic district of Orissa, India. AB - The Global Program to Eliminate Lymphatic Filariasis (GPELF), which includes alleviation of disability and suffering of patients, is run primarily in India by the primary health care system. The present study assessed the knowledge and practices related to lymphedema care among peripheral health workers of the primary health care system in a filarial-endemic district of Orissa, India. A total of 41 health workers sampled across the district were subjected to in-depth interviews. The results showed that many lymphedema patients visit the peripheral health institutions mostly for the treatment of acute episodes of lymphangitis. Many health workers do not know the concept of foot care and its importance in lymphedema management. However, a few health workers advised the patients to follow some components of foot care. The knowledge levels and practices of peripheral health workers are not at desirable levels. The medical and paramedical staff of the peripheral health institutions should be oriented about the management of lymphedema and peripheral health workers should promote the foot care practices. For the GPELF as a whole to prove successful, the patients who already have lymphedema need to be cared for and have their morbidity relieved as much as possible. PMID- 15827282 TI - Genetic structure of Aedes aegypti populations in Thailand using mitochondrial DNA. AB - A hierarchical population genetic study was conducted among 19 Aedes aegypti populations in Thailand from Chiang Mai in the north to Songkhla province in the south. Single-strand conformation polymorphism analysis was used to examine variation in a 359-basepair region of the NADH dehydrogenase subunit 4 mitochondrial DNA gene (ND4). Seven haplotypes were detected in two lineages previously identified in ND4 haplotypes from North America. Gene flow estimates and highly significant variation among populations within 25 kilometers implicated genetic drift and vector control efforts as major factors in genetic structure. Mantel regression analysis demonstrated no isolation by distance. Urban areas were relatively panmictic, while suburban/rural sites exhibited more restricted gene flow. Significant genetic structure among groups of collections > 100 kilometers apart is consistent with recent (approximately 50 year) expansion of Ae. aegypti from highly populated areas accompanied by founder effects, but could also reflect the overall low genetic diversity in ND4 in Thailand. PMID- 15827283 TI - Prevention of leprosy using rifampicin as chemoprophylaxis. AB - An intervention study was implemented on five Indonesian islands highly endemic for leprosy to determine whether rifampicin can be used as chemoprophylaxis to prevent leprosy. The population was actively screened before the intervention and subsequently once a year for three years. In the control group, no chemoprophylaxis was given. In the contact group, chemoprophylaxis was only given to contacts of leprosy patients and in the blanket group to all eligible persons. The cohort consisted of 3,965 persons. The yearly incidence rate in the control group was 39/10,000; the cumulative incidence after three years was significantly lower in the blanket group (P = 0.031). No difference was found between the contact and the control groups (P = 0.93). Whether this apparent reduced leprosy incidence in the first three years in the blanket group is due to a delayed development of leprosy or a complete clearance of infection needs to be determined. PMID- 15827284 TI - Reliability and validity of the Buruli ulcer functional limitation score questionnaire. AB - The reliability and validity of the earlier developed Buruli ulcer functional limitation score (BUFLS) questionnaire was assessed. Of 638 former Buruli ulcer patients (of 678 individuals examined), sufficient items on daily activities (>or= 13 of the 19) were applicable to calculate a score. To determine the validity, the functional limitation scores of the 638 individuals were compared with the global impression of the limitations, range of motion (ROM), and the social impact (change of occupation or education) of Buruli ulcer. To determine inter-observer reliability, the functional limitation score was reassessed in 107 participants within one and three weeks after the first interview by another interviewer and interpreter. Both global impression and ROM correlated well with the functional limitation scores (rho = 0.66 and rho = 0.61). The inter-observer reliability of 107 participants as measured by an intra-class correlation coefficient of 0.86 was very good. The functional limitation scores measured in the second assessment were significantly higher than in the first assessment. This should be taken into account when the functional limitation score is used for the individual patient. The BUFLS can be used as for between group comparisons of endpoints in clinical trials and in the planning of resources. PMID- 15827285 TI - Prevalence of Bartonella henselae in cats in Catalonia, Spain. AB - Bartonella henselae, an emerging pathogen bacterium, is the main causative agent of the cat scratch disease. While the first clinical descriptions were associated with immunosupressed patients, it is now more frequently observed in patients with normal immune status (endocarditis and bacteremia). Cats were found to be the only known reservoir of B. henselae. In this paper, we report the results obtained in the first study made to investigate the prevalence of B. henselae bacteremia and antibodies in domestic cats in Catalonia, Spain. Serum samples from 115 cats were tested for antibodies to B. henselae by immunofluorescent antibody testing, and 29.6% had a titer >or= 1:64. Seven B. henselae strains were isolated using standard culture techniques and amplification by a polymerase chain reaction and subsequent sequencing was performed on the intergenic spacer region between the 16 and 23S ribosomal RNA genes. Of all factors concerning the studied bacteremia rate (age, sex, habitat, presence of antibodies, contact with animals, parasites), only the presence of antibodies to B. henselae was statistically significant. PMID- 15827286 TI - Induction of protective immunity against scrub typhus with a 56-kilodalton recombinant antigen fused with a 47-kilodalton antigen of Orientia tsutsugamushi Karp. AB - A partial gene sequence encoding the 56-kD scrub typhus antigen (Sta56) was amplified from genomic DNA of the Orientia tsutsugamushi Karp strain by a polymerase chain reaction (PCR). The PCR product was ligated with the 47-kD scrub typhus antigen (Sta47) gene in the pQE30/47 expression vector, and the resulting recombinant expression vector was designated pQE30/56-47. A fusion antigen (Sta56 47) was expressed in Escherichia coli cells transformed with pQE30/56-47 after induction with isopropyl-beta-d-thiogalactopyranoside. The Sta56-47 antigen was recognized by both Sta47 and Sta56 immune sera and by immune serum to Sta56-47 in an immunoblot assay. This antigen was purified and used to immunize BALB/c mice. The animals immunized with Sta56-47 exhibited profound humoral and cellular immune responses, as well as increased resistance to O. tsutsugamushi Karp compared with mice immunized with Sta56 or Sta47. These results strongly suggest that Sta56-47 contains antigenic epitopes of the Sta56 and Sta47 antigens of O. tsutsugamushi Karp, and is a more suitable candidate for replacing whole-cell antigen of O. tsutsugamushi Karp to induce protective immunity against scrub typhus. PMID- 15827287 TI - Activation of cytotoxic lymphocytes in patients with scrub typhus. AB - Thai patients with scrub typhus caused by the intracellular pathogen Orientia tsutsugamushi displayed elevated plasma concentrations of granzymes A and B, interferon-gamma (IFN)-gamma-inducible protein 10, and monokine induced by IFN gamma. These data suggest that activation of cytotoxic lymphocytes is part of the early host response to scrub typhus. PMID- 15827288 TI - Seasonal effects in the elimination of trachoma. AB - The World Health Organization currently recommends annual mass antibiotic treatment to eliminate the ocular chlamydia that cause blinding trachoma. Active trachoma is believed to be seasonal in many areas of the world, and the optimal season in which to treat has not as yet been established. Here we use mathematical models of disease transmission to demonstrate that ideally, treatment should be administered before the low season to have the greatest chance of locally eliminating infection. PMID- 15827289 TI - Crimean-Congo hemorrhagic fever outbreak in Rawalpindi, Pakistan, February 2002: contact tracing and risk assessment. AB - A 25-year-old woman, later identified as index case of Crimean-Congo hemorrhagic fever (CCHF), presented to Holy Family Hospital in Rawalpindi, Pakistan with fever and generalized coagulopathy. A retrospective contact tracing was conducted to explore the modes of exposure possibly associated with transmission of CCHF infection among contacts. We traced 32 contacts of the index case and 158 contacts of secondary cases and tested them for IgG and IgM antibodies against CCHF virus by an enzyme-linked immunosorbent assay technique. According to the type of exposure, contacts were divided into five subsets: percutaneous contact with blood, blood contact to unbroken skin, cutaneous contact to non-sanguineous body fluids, physical contact with patients without body fluids contact, and close proximity without touching. Two out of four contacts who reported percutaneous exposure tested positive for antibodies to CCHF virus. We conclude that simple barrier methods and care in provision of CCHF cases may prevent transmission of this infection. PMID- 15827290 TI - Duration of infectivity and RNA of Venezuelan equine encephalitis, West Nile, and yellow fever viruses dried on filter paper and maintained at room temperature. AB - Samples of laboratory propagated Venezuelan equine encephalitis (VEE), West Nile (WN), and yellow fever (YF) viruses were blotted onto filter paper discs, air dried, and stored at room temperature. At regular intervals over a 90-day period, the dried virus samples were eluted, tested for infectivity by culture and titration in Vero cells, and examined for viral RNA by a reverse transcriptase polymerase chain reaction. The VEE, WN, and YF viral RNA was detected throughout the 90-day period in all samples examined. Infectious VEE virus could be recovered for up to 40 days; WN and YF viruses were cultured in Vero cells for up to 60 and 90 days, respectively. The results of this study demonstrate that viral nucleic acids and infectious virus can be recovered from arbovirus samples air dried on filter paper and stored at room temperature for a month or more after collection. This procedure offers a simple and inexpensive method for collecting arbovirus field specimens and transporting them to diagnostic laboratories. PMID- 15827291 TI - Acute disseminated encephalomyelitis following Plasmodium falciparum malaria caused by varicella zoster virus reactivation. AB - Neurologic complications in the course of Plasmodium falciparum infections are commonly diagnosed as cerebral malaria, but bacterial or viral meningitis may exhibit similar symptoms. One to three weeks after P. falciparum malaria, acute disseminated encephalomyelitis (ADEM) can also mimick the symptoms of cerebral malaria. We describe a 31-year-old woman with life-threatening ADEM five days after successful treatment of P. falciparum malaria. The detection of IgG and IgM antibodies in serum and cerebrospinal fluid (CSF) against multiple viruses and bacteria reflected a non-specific polyclonal B cell activation and was more confusing than helpful for diagnostic decisions. Varicella zoster virus was identified with a reverse transcriptase multiplex polymerase chain reaction in the initially obtained and frozen CSF. This case and findings from the literature indicate that P. falciparum-associated ADEM might not be immune mediated, but of infectious origin. With unclear cerebral complications during or after P. falciparum malaria, prompt initiation of empirical antiviral and antibacterial treatment in addition to antimalarials may reduce mortality. PMID- 15827292 TI - Antibody response to Culex tarsalis salivary gland antigens among sentinel chickens in California. AB - The arboviral surveillance program in California depends in part on sentinel chickens to detect western equine encephalomyelitis virus, St. Louis encephalitis virus, and West Nile virus activity. From 2000 through 2002, 1,578 serum specimens from 34 sentinel flocks in northern and southern California were tested for antibodies to Culex tarsalis salivary gland antigens. Sentinel chickens that were seropositive for mosquito salivary gland antigens were more likely to seroconvert to St. Louis encephalitis virus than those seronegative for salivary gland antigens. Flocks with mosquito traps located < 50 feet away had a reduced antibody response to mosquito salivary gland antigens. The use of sentinel chickens and mosquito traps for arboviral surveillance should be standardized to ensure that surveillance data from different sites are comparable and that flocks have comparable opportunities for mosquito exposure. Sentinel chickens should be accessible to potential mosquito vectors to maximize their sensitivity for detecting arboviral activity. PMID- 15827293 TI - A large outbreak of probable rotavirus in Nusa Tenggara Timur, Indonesia. AB - An outbreak of acute diarrheal disease was reported in Kupang, Nusa Tenggara, Indonesia, in August 2002. An investigative team carried out a retrospective historical review of records, and a case-control study involving data and specimen collections. Etiologic determination involving stool specimens was based on an enzyme-linked immunosorbent assay, with a reverse transcriptase-polymerase chain reaction performed for serotyping purposes. Two thousand six hundred probable cases were identified from hospital records during the outbreak months of June, July, August, and September 2002. Previous enteric outbreaks were recognized from the same months in the preceding years and all annual outbreak episodes following a period of prolonged, low rainfall. In contrast to previous outbreaks discerned from trend analysis, the overwhelming burden of disease fell upon the pediatric population versus the young and old in previous outbreak instances. Rotavirus was found to be the causative etiology, with serotype 1 predominating. PMID- 15827294 TI - Seroincidence and phylogeny of human immunodeficiency virus infections in a cohort of commercial sex workers in Montevideo, Uruguay. AB - A cohort study involving 60 human immunodeficiency virus (HIV)-negative male transvestite commercial sex workers (CSWs) was conducted in Montevideo, Uruguay in 1999-2001. Serum samples were tested for HIV by an enzyme-linked immunosorbent assay screening with immunoblot confirmation. Six participants seroconverted for an incidence-density rate of 6.03 (95% confidence interval = 2.21-13.12) per 100 person-years. Inconsistent condom use during client sex (adjusted hazard ratio [AHR] = 6.7), during oral sex (AHR = 5.6), and at the last sexual encounter (AHR = 7.8), and use of marihuana (AHR = 5.4) were marginally associated with HIV seroconversion. Five samples were genotyped in the protease and reverse transcriptase regions; three were subtypes B and two were BF recombinants. Full genome analysis of four samples confirmed all three subtype B samples and one of the two BF recombinants. Male transvestite CSWs sustained a high rate of HIV infection. Larger prospective studies are required to better define subtypes and associated sexual and drug-related risk factors. PMID- 15827295 TI - In the rostral ventrolateral medulla, the 70-kDa heat shock protein (HSP70), but not HSP90, confers neuroprotection against fatal endotoxemia via augmentation of nitric-oxide synthase I (NOS I)/protein kinase G signaling pathway and inhibition of NOS II/peroxynitrite cascade. AB - Heat shock proteins (HSPs) represent a group of highly conserved intracellular proteins that participate in protective adaptation against cellular stress. We evaluated the neuroprotective role of the 70-kDa HSP (HSP70) and the 90-kDa HSP (HSP90) at the rostral ventrolateral medulla (RVLM), the medullary origin of sympathetic vasomotor tone, during fatal endotoxemia. In Sprague-Dawley rats maintained under propofol anesthesia, Escherichia coli lipopolysaccharide (30 mg/kg, i.v.) induced a decrease (phase I), followed by an increase (phase II; "pro-life" phase) and a secondary decrease (phase III; "pro-death" phase) in the power density of the vasomotor component of systemic arterial pressure spectrum, along with progressive hypotension or bradycardia. Proteomic and Western blot analyses revealed that whereas HSP70 expression in the RVLM was significantly augmented during phases I and II and returned to baseline during phase III endotoxemia, HSP90 protein expression remained constant. The increase in HSP70 level was significantly blunted on pretreatment with microinjection of the transcription inhibitor actinomycin D or protein synthesis inhibitor cycloheximide into the bilateral RVLM. Functional blockade of HSP70 in the RVLM by an anti-HSP70 antiserum or prevention of synthesis by an antisense hsp70 oligonucleotide exacerbated mortality or potentiated the cardiovascular depression during experimental endotoxemia, alongside significantly reduced nitric-oxide synthase (NOS) I or protein kinase G (PKG) level or augmented NOS II or peroxynitrite level in the RVLM. We conclude that whereas HSP90 is ineffective, de novo synthesis of HSP70 in the RVLM may confer neuroprotection during fatal endotoxemia by preventing cardiovascular depression via enhancing the sympathoexcitatory NOS I/PKG signaling pathway and inhibiting the sympathoinhibitory NOS II/peroxynitrite cascade in the RVLM. PMID- 15827296 TI - The cover. Calculating. PMID- 15827297 TI - Magnetism on the brain: researchers probe transcranial magnetic stimulation. PMID- 15827298 TI - Gene-tweaking animal studies seen as advances for restoring hearing, vision. PMID- 15827299 TI - Kidney donor exchange program planned. PMID- 15827300 TI - Lori Mosca, MD, PhD: raising awareness of heart disease risks in women. Interview by Mike Mitka. PMID- 15827304 TI - Trends in cardiovascular complications of diabetes. PMID- 15827305 TI - Lead exposure and cataract risk in men. PMID- 15827306 TI - Lead exposure and cataract risk in men. PMID- 15827307 TI - Health risks associated with polychlorinated biphenyls. PMID- 15827308 TI - Systematic review of bariatric surgery. PMID- 15827309 TI - Systematic review of bariatric surgery. PMID- 15827310 TI - Super Bowls: serving bowl size and food consumption. PMID- 15827311 TI - Racial differences in the use of BRCA1/2 testing among women with a family history of breast or ovarian cancer. AB - CONTEXT: Given the current context of racial disparities in health and health care and the historical context of eugenics, racial disparities in the use of genetic susceptibility testing have been widely anticipated. However, to our knowledge there are no published studies examining the magnitude and determinants of racial differences in the use of genetic susceptibility testing. OBJECTIVES: To investigate the relationship between race and the use of BRCA1/2 counseling among women with a family history of breast or ovarian cancer and to determine the contribution of socioeconomic characteristics, cancer risk perception and worry, attitudes about genetic testing, and interactions with primary care physicians to racial differences in utilization. DESIGN, SETTING, AND PARTICIPANTS: Case-control study (December 1999-August 2003) of 408 women with a family history of breast or ovarian cancer, of whom 217 underwent genetic counseling for BRCA1/2 testing (cases) and 191 women did not (controls). Participants received primary care within a large health system in greater Philadelphia, Pa. MAIN OUTCOME MEASURES: Probability of carrying a BRCA1/2 mutation, socioeconomic characteristics, perception of breast and ovarian cancer risk, worry about breast and ovarian cancer, attitudes about BRCA1/2 testing, and primary care physician discussion of BRCA1/2 testing were measured prior to undergoing BRCA1/2 counseling for cases and at the time of enrollment for controls. RESULTS: African American women with a family history of breast or ovarian cancer were significantly less likely to undergo genetic counseling for BRCA1/2 testing than were white women with a family history of breast or ovarian cancer (odds ratio, 0.22; 95% confidence interval, 0.12-0.40). This association persisted after adjustment for probability of BRCA1/2 mutation, socioeconomic characteristics, breast and ovarian cancer risk perception and worry, attitudes about the risks and benefits of BRCA1/2 testing, and primary care physician discussion of BRCA1/2 testing (adjusted odds ratio for African American vs white, 0.28; 95% confidence interval, 0.09-0.89). CONCLUSIONS: Racial disparities in the use of BRCA1/2 counseling are large and do not appear to be explained by differences in risk factors for carrying a BRCA1/2 mutation, socioeconomic factors, risk perception, attitudes, or primary care physician recommendations. The benefit of predictive genetic testing will not be fully realized unless these disparities can be addressed. PMID- 15827312 TI - Cardiovascular mortality risk in chronic kidney disease: comparison of traditional and novel risk factors. AB - CONTEXT: Elderly persons with chronic kidney disease have substantial risk for cardiovascular mortality, but the relative importance of traditional and novel risk factors is unknown. OBJECTIVE: To compare traditional and novel risk factors as predictors of cardiovascular mortality. DESIGN, SETTING, AND PATIENTS: A total of 5808 community-dwelling persons aged 65 years or older living in 4 communities in the United States participated in the Cardiovascular Health Study cohort. Participants were initially recruited from 1989 to June 1990; an additional 687 black participants were recruited in 1992-1993. The average length of follow-up in this longitudinal study was 8.6 years. MAIN OUTCOME MEASURES: Cardiovascular mortality among those with and without chronic kidney disease. Chronic kidney disease was defined as an estimated glomerular filtration rate of less than 60 mL/min per 1.73 m2. RESULTS: Among the participants, 1249 (22%) had chronic kidney disease at baseline. The cardiovascular mortality risk rate was 32 deaths/1000 person-years among those with chronic kidney disease vs 16/1000 person-years among those without it. In multivariate analyses, diabetes, systolic hypertension, smoking, low physical activity, nonuse of alcohol, and left ventricular hypertrophy were predictors of cardiovascular mortality in persons with chronic kidney disease (all P values <.05). Among the novel risk factors, only log C-reactive protein (P = .05) and log interleukin 6 (P<.001) were associated with the outcome as linear predictors. Traditional risk factors were associated with the largest absolute increases in risks for cardiovascular deaths among persons with chronic kidney disease: for left ventricular hypertrophy, there were 25 deaths per 1000 person-years; current smoking, 20 per 1000 person years; physical inactivity, 15 per 1000 person-years; systolic hypertension, 14 per 1000 person-years; diabetes, 14 per 1000 person-years; and nonuse of alcohol, 11 per 1000 person-years vs 5 deaths per 1000 person-years for those with increased C-reactive protein and 5 per 1000 person-years for those with increased interleukin 6 levels. A receiver operating characteristic analysis found that traditional risk factors had an area under the curve of 0.73 (95% confidence interval, 0.70-0.77) among those with chronic kidney disease. Adding novel risk factors only increased the area under the curve to 0.74 (95% confidence interval, 0.71-0.78; P for difference = .15). CONCLUSIONS: Traditional cardiovascular risk factors had larger associations with cardiovascular mortality than novel risk factors in elderly persons with chronic kidney disease. Future research should investigate whether aggressive lifestyle intervention in patients with chronic kidney disease can reduce their substantial cardiovascular risk. PMID- 15827313 TI - Relationship of incorrect dosing of fibrinolytic therapy and clinical outcomes. AB - CONTEXT: Incorrect dosing of alteplase has been associated with worse clinical outcomes in patients. However, patients at high risk of adverse events are more prone to dosing errors, thus confounding this relationship. OBJECTIVE: To determine if the association between incorrect dosing of alteplase and adverse outcomes is related to cause and effect or to confounding. DESIGN, SETTING, AND PATIENTS: Observational analysis in May 2004 of a double-blind, double-dummy trial of 16,949 patients with ST-segment elevation myocardial infarction who were enrolled in the Assessment of the Safety and Efficacy of a New Thrombolytic (ASSENT-2) trial and were assigned to either a bolus of tenecteplase (with alteplase placebo bolus plus infusion) or a bolus of alteplase (with tenecteplase placebo plus infusion). MAIN OUTCOME MEASURES: Thirty-day mortality, in-hospital stroke, and major bleeding associated with incorrect dosing of active alteplase compared with placebo alteplase. RESULTS: Incorrect dosing occurred in 4.9% of patients who received active alteplase and in 4.6% of patients who received alteplase placebo. Patients receiving incorrect doses of alteplase or alteplase placebo were more likely to be older, female, black, shorter, have lower body weight and systolic blood pressure, and have a higher Killip class at presentation. Thirty-day mortality was higher in patients who received an overdose (9.8%) or underdose (19.5%) of alteplase compared with those who received a correct dose (5.4%). The same pattern was present in patients who received an alteplase placebo (10.0% for overdose, 23.5% for underdose, and 5.4% for correct dose). Similar patterns were seen for in-hospital intracranial hemorrhage and major bleeding. The higher rates of adverse outcomes with incorrect dosing were largely accounted for by adjusting for baseline characteristics. CONCLUSIONS: The relationship between incorrect dosing and patient outcome in ASSENT-2 is primarily due to confounding factors rather than incorrect dosing itself. These data highlight the need for caution when ascribing a causal relationship to associations between incorrect dosing and adverse outcomes. PMID- 15827314 TI - Immunogenicity and safety of a combination pneumococcal-meningococcal vaccine in infants: a randomized controlled trial. AB - CONTEXT: The success of conjugate vaccines in decreasing invasive disease due to Streptococcus pneumoniae and group C Neisseria meningitidis has placed pressure on crowded infant immunization schedules, making development of combination vaccines a priority. OBJECTIVE: To determine the safety and immunogenicity of a combination 9-valent pneumococcal-group C meningococcal conjugate candidate vaccine (Pnc9-MenC) administered as part of the routine UK infant immunization schedule at ages 2, 3, and 4 months. DESIGN, SETTING, AND PARTICIPANTS: Phase 2 randomized controlled trial conducted from August 2000 to January 2002 and enrolling 240 healthy infants aged 7 to 11 weeks from 2 UK centers, with home follow-up visits at ages 2, 3, 4, and 5 months. INTERVENTION: Pnc9-MenC (n = 120) or monovalent group C meningococcal conjugate vaccine (MenC) (n = 120) administered in addition to routine immunizations (diphtheria and tetanus toxoids and whole-cell pertussis [DTwP], Haemophilus influenzae type b [Hib] polyribosylribitol phosphate-tetanus toxoid protein conjugate, oral polio vaccine). MAIN OUTCOME MEASURES: Group C meningococcal immunogenicity measured by serum bactericidal titer (SBT) 1 month following the third dose; rates of postimmunization reactions. RESULTS: MenC component immunogenicity was reduced in the Pnc9-MenC vs the MenC group (geometric mean SBT, 179 [95% confidence interval {CI}, 133-243] vs 808 [95% CI, 630-1037], respectively; P<.001). The proportion with group C meningococcal SBT greater than 1:8 was lower in the Pnc9-MenC vs the MenC group (95% vs 100%, P = .05). The geometric mean concentration of antibodies to concomitantly administered Hib vaccine was reduced in the Pnc9-MenC vs the MenC group (2.11 [95% CI, 1.57-2.84] microg/mL vs 3.36 [95% CI, 2.57-4.39] microg/mL; P = .02), as were antibodies against diphtheria (0.74 [95% CI, 0.63 0.87] microg/mL vs 1.47 [95% CI, 1.28-1.69] microg/mL; P<.001). Pnc9-MenC was immunogenic for each of 9 contained pneumococcal serotypes, with responses greater than 0.35 microg/mL observed in more than 88% of infants. Increased irritability and decreased activity were observed after the third dose in the Pnc9-MenC group. CONCLUSIONS: Pnc9-MenC combination vaccine administered to infants at ages 2, 3, and 4 months demonstrated reduced group C meningococcal immunogenicity compared with MenC vaccine. The immunogenicity of concomitantly administered Hib and DTwP vaccines was also diminished. The Pnc9-MenC vaccine was safe and immunogenic for all contained pneumococcal serotypes. The reduced MenC immunogenicity may limit the development of the Pnc9-MenC vaccine. PMID- 15827315 TI - Abciximab as adjunctive therapy to reperfusion in acute ST-segment elevation myocardial infarction: a meta-analysis of randomized trials. AB - CONTEXT: The benefits of abciximab in patients with ST-segment elevation myocardial infarction (STEMI) are still a matter of debate. OBJECTIVE: To combine data from all randomized trials conducted with abciximab in STEMI. DATA SOURCES: Formal searches of electronic databases (MEDLINE, PubMed) from from January 1990 to December 2004. STUDY SELECTION: We examined all completed, published, randomized trials of abciximab in STEMI. The following key words were used for study selection: randomized trial, myocardial infarction, reperfusion, primary angioplasty, facilitated angioplasty, stenting, fibrinolysis, IIb-IIIa inhibitors, and abciximab. DATA EXTRACTION: Information on study design, type and dosage of drugs, inclusion and exclusion criteria, number of patients, and clinical outcome was extracted by 2 investigators. Disagreements were resolved by consensus. DATA SYNTHESIS: Eleven trials were analyzed, involving 27115 patients (12,602 [46.5%] in the abciximab group, 14,513 [53.5%] in the control group). When compared with the control group, abciximab was associated with a significant reduction in short-term (30 days) mortality (2.4% vs 3.4%, P = .047) and long term (6-12 months) mortality (4.4% vs 6.2%, P = .01) in patients undergoing primary angioplasty but not in those treated with fibrinolysis or in all trials combined. Abciximab was associated with a significant reduction in 30-day reinfarction, both in all trials combined (2.1% vs 3.3%, P<.001), in primary angioplasty (1.0% vs 1.9%, P = .03), and in fibrinolysis trials (2.3% vs 3.6%, P<.001). Abciximab did not result in an increased risk of intracranial bleeding (0.61% vs 0.62%, P = .62) but was associated with an increased risk of major bleeding complications when combined with fibrinolysis (5.2% vs 3.1%, P<.001) but not with primary angioplasty (4.7% vs 4.1%, P = .36). CONCLUSIONS: This meta analysis shows that, when compared with the control group, adjunctive abciximab for STEMI is associated with a significant reduction in 30-day and long-term mortality in patients treated with primary angioplasty but not in those receiving fibrinolysis. The 30-day reinfarction rate is significantly reduced in patients treated with either fibrinolysis or primary angioplasty. A higher risk of major bleeding complications is observed with abciximab in association with fibrinolysis. PMID- 15827316 TI - HIPAA and patient care: the role for professional judgment. AB - Federal health privacy regulations, commonly known as the Health Insurance Portability and Accountability Act (HIPAA) regulations, came into effect in April 2003. Many clinicians and institutions have relied on consultants and risk managers to tell them how to implement these regulations. Much of the controversy and confusion over the HIPAA regulations concern so-called incidental disclosures. Some interpretations of the privacy regulations would limit essential communication and compromise good patient care. This article analyzes misconceptions regarding what the regulations say about incidental disclosures and discusses the reasons for such misunderstandings. Many misconceptions arise from gaps in the regulations. These gaps are appropriately filled by professional judgment informed by ethical guidelines. The communication should be necessary and effective for good patient care, and the risks of a breach of confidentiality should be proportional to the likely benefit for the patient's care. The alternative for communication should be impractical. We offer specific recommendations to help physicians think through what incidental disclosures in patient care are ethically permissible and what safeguards ought to be taken. Physicians should work with risk managers and practice administrators to develop policies that promote good communication in patient care, while taking appropriate steps to protect patient privacy. PMID- 15827317 TI - A 64-year-old woman with primary hyperparathyroidism. PMID- 15827318 TI - A 60-year-old woman trying to discontinue hormone replacement therapy, 2 years later. PMID- 15827319 TI - How healthy are US children? PMID- 15827320 TI - Confronting genetic testing disparities: knowledge is power. PMID- 15827321 TI - JAMA patient page. Hyperparathyroidism. PMID- 15827322 TI - Teaching resources. G-protein-coupled receptors. AB - This Teaching Resource provides lecture notes and slides for a class covering G protein-coupled receptors (GPCRs) and is part of the course "Cell Signaling Systems: A Course for Graduate Students." The lecture begins with a discussion of the major classes of GPCRs and then proceeds to describe the mechanisms of receptor diversity, ligand interaction, desensitization, coupling, and mutations associated with human diseases. PMID- 15827323 TI - Small-interfering RNA-induced androgen receptor silencing leads to apoptotic cell death in prostate cancer. AB - Prostate cancer is the second leading cause of cancer death in the United States and, thus far, there has been no effective therapy for the treatment of hormone refractory disease. Recently, the androgen receptor (AR) has been shown to play a critical role in the development and progression of the disease. In this report, we showed that knocking down the AR protein level by a small interfering RNA (siRNA) approach resulted in a significant apoptotic cell death as evidenced by an increased annexin V binding, reduced mitochondrial potential, caspase-3/6 activation, and DFF45 and poly(ADP-ribose) polymerase cleavage. The apoptotic response was specifically observed in those siRNA-transfected cells that harbor a native AR gene. No cell death was found in the AR-null prostate cancer cell PC-3 or its subline that has been reconstituted with an exogenous AR gene, as well as two breast cancer cell lines that are AR positive. Moreover, in parallel with the siRNA-induced AR silencing, the antiapoptotic protein Bcl-xL was significantly reduced, which might account for the apoptotic cell death because ectopic enforced expression of Bcl-xL protein partially inhibited apoptosis after AR silencing. Taken together, our data showed that knocking down the AR protein level in prostate cancer cells leads to apoptosis by disrupting the Bcl-xL mediated survival signal downstream of AR-dependent survival pathway. PMID- 15827324 TI - RNA expression profiling of normal and tumor cells following photodynamic therapy with 5-aminolevulinic acid-induced protoporphyrin IX in vitro. AB - Photodynamic therapy using 5-aminolevulinic acid-induced protoporphyrin IX synthesis as a photosensitizing reagent is an encouraging modality for cancer treatment. Understanding the mechanism of tumor phototoxicity is important to provide a basis for combinatory therapy regimens. A normal cell line (UROtsa, urothelial) and two tumor cell lines (RT4, urothelial; HT29, colonic) were treated with cell line-specific LD50 doses of light after exposure to 5 aminolevulinic acid (100 microg/mL), and harvested for RNA extraction 0, 10, and 30 minutes after irradiation. The RNA was hybridized to the metg001A Affymetrix GeneChip containing 2,800 genes, focusing on cancer-related and growth regulatory targets. Comparing the gene expression profiles between the different samples, 40 genes (e.g., SOD2, LUC7A, CASP8, and DUSP1) were identified as significantly altered in comparison with the control samples, and grouped according to their gene ontology. We selected caspase-8 (CASP8) and dual specificity phosphatase 1 (DUSP1) for further validation of the array findings, and compared their expression with the expression of the immediate early gene FOS by quantitative reverse transcription-PCR. RNA expression of CASP8 stayed unchanged whereas DUSP1 RNA was up-regulated in normal and tumor cells starting 30 minutes after irradiation. In contrast, FOS RNA was found continuously up-regulated over time in all three cell lines. Induction of DUSP1 protein expression was clearly shown after 1 hour using Western blot analysis. Interestingly, no changes of caspase-8 protein expression but activation of catalytic activity was detected only in UROtsa cells starting 1 hour after photodynamic therapy, whereas no changes were seen in both tumor cell lines. According to caspase-8, the active caspase 3 fragment was found only in the normal urothelial cell line (UROtsa) 1 hour after photodynamic therapy. Combined data analysis suggests that photodynamic therapy in vitro (LD50) leads to apoptosis in UROtsa and to necrosis in the tumor cell lines, respectively. RNA expression profiling of normal and tumor cell lines following photodynamic therapy with 5-aminolevulinic acid gave insight into the major molecular mechanisms induced by photodynamic therapy. PMID- 15827325 TI - Ku protein targeting by Ku70 small interfering RNA enhances human cancer cell response to topoisomerase II inhibitor and gamma radiation. AB - Ku protein is a heterodimer (Ku70 and Ku86) known to play an important role in V(D)J recombination, apoptosis, telomere fusion, and double-strand break repair. Its role in double-strand breaks is relevant to cancer therapy because lack of Ku86 causes one of the most radiation-responsive phenotypes (hamster cells, XRS5). Although it is known that the heterodimer is necessary for the various functions of this protein, the impact of targeting Ku in human cancer cells has not been shown due to lack of appropriate approaches. It is also not known whether complete knock-out of Ku protein is required to enhance the sensitivity of human cells to gamma radiation as Ku protein is much more abundant in human cells than in hamster cells. In the current article, we have investigated the direct effect of Ku70 depletion in human cervical epithelioid (HeLa) and colon carcinoma (HCT116) cells. We specifically targeted Ku70 mRNA by use of small interfering RNA (siRNA). Of the five Ku70 siRNA synthesized, three inhibited the expression of Ku70 by up to 70% in HeLa cells. We have tested the effect of chemically synthesized siRNAs for target sequence 5 (CS #5) on the response of HeLa cells 72 hours after transfection to gamma radiation and etoposide, as this showed the maximum inhibition of Ku70 expression. Ku70 siRNA induced a decrease in the surviving fraction of irradiated HeLa cells by severalfold. Similar sensitizing effects were observed for etoposide, a topoisomerase II inhibitor. Studies with HCT116 cells using the same Ku70 siRNA (CS #5) showed a direct correlation between expression of Ku70 and sensitization to radiation and etoposide treatments. PMID- 15827326 TI - Pentameric procyanidin from Theobroma cacao selectively inhibits growth of human breast cancer cells. AB - A naturally occurring, cocoa-derived pentameric procyanidin (pentamer) was previously shown to cause G0/G1 cell cycle arrest in human breast cancer cells by an unknown molecular mechanism. Here, we show that pentamer selectively inhibits the proliferation of human breast cancer cells (MDA MB-231, MDA MB-436, MDA MB 468, SKBR-3, and MCF-7) and benzo(a)pyrene-immortalized 184A1N4 and 184B5 cells. In contrast, normal human mammary epithelial cells in primary culture and spontaneously immortalized MCF-10A cells were significantly resistant. We evaluated whether this differential response to pentamer may involve depolarization of the mitochondrial membrane. Pentamer caused significant depolarization of mitochondrial membrane in MDA MB231 cells but not the more normal MCF-10A cells, whereas other normal and tumor cell lines tested gave variable results. Further investigations, using a proteomics approach with pentamer-treated MDA MB-231, revealed a specific dephosphorylation, without changes in protein expression, of several G1-modulatory proteins: Cdc2 (at Tyr15), forkhead transcription factor (at Ser256, the Akt phosphorylation site) and p53 (Ser392). Dephosphorylation of p53 (at Ser392) by pentamer was confirmed in MDA MB-468 cells. However, both expression and phosphorylation of retinoblastoma protein were decreased after pentamer treatment. Our results show that breast cancer cells are selectively susceptible to the cytotoxic effects of pentameric procyanidin, and suggest that inhibition of cellular proliferation by this compound is associated with the site-specific dephosphorylation or down regulation of several cell cycle regulatory proteins. PMID- 15827327 TI - Expression of multidrug transporter MRP4/ABCC4 is a marker of poor prognosis in neuroblastoma and confers resistance to irinotecan in vitro. AB - Members of the multidrug resistance-associated protein (MRP) family of transporters are believed to contribute to cytotoxic drug resistance and chemotherapy failure. We observed frequent MRP4 overexpression in aggressive primary neuroblastoma, a disease for which we have previously shown MRP1 to be a prognostic indicator. High MRP4 expression correlated with MYCN oncogene amplification and was significantly associated with poor clinical outcome. Although MRP4 is known to transport some nucleoside analogues, it has not previously been associated with resistance to drugs used to treat solid tumors. We now show that it mediates substantial resistance in vitro to the topoisomerase I poison irinotecan/CPT-11 and its active metabolite SN-38. These results suggest that MRP4 will be a useful prognostic marker for neuroblastoma and that clinical trials of irinotecan as a neuroblastoma treatment should monitor MRP4 expression. The same may be true for other tumor types expressing high levels of the transporter. PMID- 15827328 TI - Resveratrol-induced apoptotic death in human U251 glioma cells. AB - Resveratrol (trans-3,4',5-trihydroxystilbene) is a naturally occurring polyphenolic compound highly enriched in grapes, peanuts, red wine, and a variety of food sources. Resveratrol has antiinflammatory and antioxidant properties, and also has potent anticancer properties. Human glioma U251 cells were used to understand the molecular mechanisms by which resveratrol acts as an anticancer agent, since glioma is a particularly difficult cancer to treat and eradicate. Our data show that resveratrol induces dose- and time-dependent death of U251 cells, as measured by lactate dehydrogenase release and internucleosomal DNA fragmentation assays. Resveratrol induces activation of caspase-3 and increases the cleavage of the downstream caspase substrate, poly(ADP-ribose) polymerase. Resveratrol-induced DNA fragmentation can be completely blocked by either a general caspase inhibitor (Z-VAD-FMK) or a selective caspase-3 inhibitor (Z-DEVD FMK), but not by a selective caspase-1 inhibitor. Resveratrol induces cytochrome c release from mitochondria to the cytoplasm and activation of caspase-9. Resveratrol also increases expression of proapoptotic Bax and its translocation to the mitochondria. Resveratrol inhibits U251 proliferation, as measured by MTS assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl) 2H-tetrazolium, inner salt], and induces G0/G1 growth arrest, as determined by flow cytometry. The cyclin-dependent kinase inhibitor, olomoucine, prevents cell cycle progression and resveratrol-induced apoptosis. These results suggest that multiple signaling pathways may underlie the apoptotic death of U251 glioma induced by resveratrol, which warrants further exploration as an anticancer agent in human glioma. PMID- 15827329 TI - A-432411, a novel indolinone compound that disrupts spindle pole formation and inhibits human cancer cell growth. AB - Microtubules are among the most successful targets for anticancer therapies and for the development of new anticancer drugs. A-432411 is a novel small molecule that destabilizes microtubules at high concentration and disrupts normal spindle formation at low concentration. A-432411 is an indolinone that is structurally different from other known synthetic microtubule inhibitors. This compound is efficacious against a variety of human cancer cell lines including drug-resistant HCT-15 that overexpresses Pgp170. Biochemical studies show that A-432411 competes with the colchicine-binding site on tubulin and inhibits microtubule polymerization. Fluorescence-activated cell sorting analysis indicates that A 432411 causes G2-M arrest and induces apoptosis. Cells treated with A-432411 have increased level of phospho-histone H3 at Ser10 and decreased level of phospho cdc2 at Tyr15. Concurrently, securin and cyclin B1 expression levels remain the same, indicating the activation of the spindle checkpoint. Immunocytochemistry and fluorescence microscopy experiments reveal that 1 micromol/L A-432411 destabilizes microtubules in cells. At 0.1 micromol/L, the compound disrupts normal spindle pole formation possibly through stabilization of microtubule dynamic. Both structural and cellular properties of A-432411 make it an attractive candidate for further development. PMID- 15827330 TI - 8-Amino-adenosine induces loss of phosphorylation of p38 mitogen-activated protein kinase, extracellular signal-regulated kinase 1/2, and Akt kinase: role in induction of apoptosis in multiple myeloma. AB - Multiple myeloma is a slowly proliferating B-cell malignancy that accumulates apoptosis-resistant and replication-quiescent cell populations, posing a challenge for current chemotherapeutics that target rapidly replicating cells. Multiple myeloma remains an incurable disease in need of new therapeutic approaches. The purine nucleoside analogue, 8-amino-adenosine (8-NH2-Ado), exhibits potent activity in preclinical studies, inducing apoptosis in several multiple myeloma cell lines. This cytotoxic effect requires phosphorylation of 8 NH2-Ado to its triphosphate form, 8-amino-ATP, and results in a concomitant loss of endogenous ATP levels. Here, we show the novel effect of 8-NH2-Ado on the phosphorylation status of key cellular signaling molecules. Multiple myeloma cells treated with 8-NH2-Ado exhibit a dramatic loss of phosphorylation of several important signaling proteins, including extracellular signal-regulated kinase 1/2, p38 mitogen-activated protein kinase, and Akt kinase. Cells depleted of ATP independent of 8-NH2-Ado do not exhibit the same decrease in phosphorylation of vital cellular proteins. Therefore, the significant shifts in endogenous ATP pools caused by 8-NH2-Ado treatment cannot account for the changes in phosphorylation levels. Instead, 8-NH2-Ado may influence the activity of select regulatory protein kinases and/or phosphatases, with preliminary data suggesting that protein phophatase 2A activity is affected by 8-NH2-Ado. The distinctive effect of 8-NH2-Ado on the phosphorylation status of cellular proteins is a novel phenomenon for a nucleoside analogue drug and is unique to 8 NH2-Ado among this class of drugs. The kinetics of 8-NH2-Ado-mediated changes in phosphorylation levels of critical prosurvival and apoptosis-regulating proteins suggests that the modulation of these proteins by dephosphorylation at early time points may be an important mechanistic step in 8-NH2-Ado-induced apoptosis. PMID- 15827331 TI - Oridonin, a diterpenoid purified from Rabdosia rubescens, inhibits the proliferation of cells from lymphoid malignancies in association with blockade of the NF-kappa B signal pathways. AB - This study found that oridonin, a natural diterpenoid purified from Rabdosia rubescens, inhibited growth of multiple myeloma (MM; U266, RPMI8226), acute lymphoblastic T-cell leukemia (Jurkat), and adult T-cell leukemia (MT-1) cells with an effective dose that inhibited 50% of target cells (ED50) ranging from 0.75 to 2.7 microg/mL. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling staining showed that oridonin caused apoptosis of MT-1 cells in a time-dependent manner. We explored effects of oridonin on antiapoptotic Bcl-2 family members and found that it down-regulated levels of Mcl-1 and BCL-x(L), but not Bcl-2 protein, in both MT-1 and RPMI8226 cells. Further studies found that oridonin inhibited nuclear factor-kappa B (NF-kappa B) DNA-binding activity in these cells as measured by luciferase reporter gene, ELISA-based, and electrophoretic mobility shift assays. Oridonin also blocked tumor necrosis factor-alpha- and lipopolysaccharide-stimulated NF-kappa B activity in Jurkat cells as well as RAW264.7 murine macrophages. Of note, oridonin decreased survival of freshly isolated adult T-cell leukemia (three samples), acute lymphoblastic leukemia (one sample), chronic lymphocytic leukemia (one sample), non-Hodgkin's lymphoma (three samples), and MM (four samples) cells from patients in association with inhibition of NF-kappa B DNA-binding activity. On the other hand, oridonin did not affect survival of normal lymphoid cells from healthy volunteers. Taken together, oridonin might be useful as adjunctive therapy for individuals with lymphoid malignancies, including the lethal disease adult T-cell leukemia. PMID- 15827332 TI - Parthenolide and sulindac cooperate to mediate growth suppression and inhibit the nuclear factor-kappa B pathway in pancreatic carcinoma cells. AB - Activation of the transcription factor nuclear factor-kappa B (NF-kappa B) has been implicated in pancreatic tumorigenesis. We evaluated the effect of a novel NF-kappa B inhibitor, parthenolide, a sesquiterpene lactone isolated from the herb feverfew, in three human pancreatic tumor cell lines (BxPC-3, PANC-1, and MIA PaCa-2). Parthenolide inhibited pancreatic cancer cell growth in a dose dependent manner with substantial growth inhibition observed between 5 and 10 micromol/L parthenolide in all three cell lines. Parthenolide treatment also dose dependently increased the amount of the NF-kappa B inhibitory protein, I kappa B alpha, and decreased NF-kappa B DNA binding activity. We have previously shown that nonsteroidal anti-inflammatory drugs (NSAID) suppress the growth of pancreatic cancer cells. To determine whether inhibition of the NF-kappa B pathway by parthenolide could sensitize pancreatic cancer cells to NSAID inhibition, BxPC-3, PANC-1, and MIA PaCa-2 cells were treated with parthenolide and the NSAID sulindac, either alone or in combination. Treatment with the combination of parthenolide and sulindac inhibited cell growth synergistically in MIA PaCa-2 and BxPC-3 cells and additively in PANC-1 cells. In addition, treatment with the parthenolide/sulindac combination lowered the threshold for apoptosis. Increased levels of I kappa B-alpha protein were detected, especially in MIA PaCa-2 cells, after treatment with parthenolide and sulindac compared with each agent alone. Similarly, decreased NF-kappa B DNA binding and transcriptional activities were detected in cells treated with the combination compared with the single agents, demonstrating cooperative targeting of the NF-kappa B pathway. These data provide preclinical support for a combined chemotherapeutic approach with NF-kappa B inhibitors and NSAIDs for the treatment of pancreatic adenocarcinoma. PMID- 15827334 TI - Histone deacetylase inhibitors induce G2-checkpoint arrest and apoptosis in cisplatinum-resistant ovarian cancer cells associated with overexpression of the Bcl-2-related protein Bad. AB - Trichostatin A produces predominantly G(1) cell-cycle blockade and differentiation of the cisplatinum-sensitive A2780 ovarian cancer cell line. Given the propensity of ovarian tumors to become resistant to cisplatinum, often leading to cross-resistance to other agents, we have extended these observations by examining how the emergence of resistant phenotypes in A2780 cells affects the actions of histone deacetylase (HDAC) inhibitors. Trichostatin A exposure (100 ng/mL, 24 hours) induced ultrastructural differentiation of the "intrinsically" cisplatinum-resistant A2780-9M subline, with the reappearance of intercellular junctions and lumina containing primitive microvilli. Similar trichostatin A exposure in the acquired resistance A2780CP cells produced minimal differentiation consisting of occasional weak intercellular junctions. Independent of the differences in trichostatin A-induced differentiation, in both resistant sublines trichostatin A produced a similar reduction in cell viability, by >90%, within 5 days of treatment. Diminished viability in both A2780-9M and CP cells was associated with the absence of cell cycle arrest in G1, resulting in predominant G2-checkpoint arrest accompanied by a 10- to 20-fold increase in Annexin V binding and the reemergence of apoptosis. Similar cell cycle arrests and apoptosis were also observed using other HDAC inhibitors and in other resistant ovarian cancer cell lines (OVCAR-3 and SK-OV-3). Trichostatin A-induced apoptosis in resistant cells is in sharp contrast to its effects on the parental cisplatinum-sensitive A2780 and normal MRC-5 fibroblast cell lines (predominant cycle arrest in G1 with no detectable apoptosis). Western immunoblot analysis indicated trichostatin A triggers apoptosis in resistant ovarian cancer cells via p53-independent activation of the intrinsic "mitochondrial" pathway, commensurate with induction of the Bcl-2-related protein Bad. These results suggest cisplatinum resistance alters the effects of HDAC inhibition through a shift in cell cycle arrest from the G1 to the G2 checkpoint and reactivation of the intrinsic mitochondrial apoptotic cascade. PMID- 15827333 TI - Novel hydroxyl naphthoquinones with potent Cdc25 antagonizing and growth inhibitory properties. AB - Cdc25 phosphatases are important in cell cycle control and activate cyclin dependent kinases (Cdk). Efforts are currently under way to synthesize specific small-molecule Cdc25 inhibitors that might have anticancer properties. NSC 95397, a protein tyrosine phosphatase antagonist from the National Cancer Institute library, was reported to be a potent Cdc25 inhibitor. We have synthesized two hydroxyl derivatives of NSC 95397, monohydroxyl-NSC 95397 and dihydroxyl-NSC 95397, which both have enhanced activity for inhibiting Cdc25s. The new analogues, especially dihydroxyl-NSC 95397, potently inhibited the growth of human hepatoma and breast cancer cells in vitro. They influenced two signaling pathways. The dual phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was induced, likely due to inhibition of the ERK phosphatase activity in Hep 3B cell lysate but not the dual specificity ERK phosphatase MKP-1. They also inhibited Cdc25 enzymatic activities and induced tyrosine phosphorylation of the Cdc25 target Cdks. Addition of hydroxyl groups to the naphthoquinone ring thus enhanced the potency of NSC 95397. These two new compounds may be useful probes for the biological functions of Cdc25s and have the potential for disrupting the cell cycle of growing tumor cells. PMID- 15827335 TI - Bovine lactoferricin selectively induces apoptosis in human leukemia and carcinoma cell lines. AB - Bovine lactoferricin (LfcinB) is a cationic, amphipathic peptide that is cytotoxic for human and rodent cancer cells. However, the mechanism by which LfcinB causes the death of cancer cells is not well understood. Here, we show that in vitro treatment with LfcinB rapidly induced apoptosis in several different human leukemia and carcinoma cell lines as determined by DNA fragmentation assays and phosphatidylserine headgroup inversion detected by Annexin V binding to the surface of cancer cells. Importantly, LfcinB treatment did not adversely affect the viability of untransformed human lymphocytes, fibroblasts, or endothelial cells. Studies with different LfcinB-derived peptide fragments revealed that the cytotoxic activity of LfcinB resided within the amino acid sequence FKCRRWQWRM. Treatment of Jurkat T leukemia cells with LfcinB resulted in the production of reactive oxygen species followed by caspase-2 induced dissipation of mitochondrial transmembrane potential and subsequent activation of caspase-9 and caspase-3. Selective inhibitors of caspase-2 (Z-VDVAD FMK), caspase-9 (Z-LEHD-FMK), and caspase-3 (Z-DEVD-FMK) protected both leukemia and carcinoma cells from LfcinB-induced apoptosis. Conversely, a caspase-8 inhibitor (Z-IETD-FMK) had no effect, which argued against a role for caspase-8 and was consistent with the finding that death receptors were not involved in LfcinB-induced apoptosis. Furthermore, Jurkat T leukemia cells that overexpressed Bcl-2 were less sensitive to LfcinB-induced apoptosis, which was characterized by mitochondrial swelling and the release of cytochrome c from mitochondria into the cytosolic compartment. We conclude that LfcinB kills cancer cells by triggering the mitochondrial pathway of apoptosis at least in part through the generation of reactive oxygen species. PMID- 15827336 TI - p53 gene therapy of human osteosarcoma using a transferrin-modified cationic liposome. AB - Gene delivery via transferrin receptors, which are highly expressed by cancer cells, can be used to enhance the effectiveness of gene therapy for cancer. In this study, we examined the efficacy of p53 gene therapy in human osteosarcoma (HOSM-1) cells derived from the oral cavity using a cationic liposome supplemented with transferrin. HOSM-1 cells were exposed to transferrin-liposome p53 in vitro, and the growth inhibition rate, expression of p53 and bax, and induction of apoptosis were measured 48 hours later. Treatment of HOSM-1 cells with transferrin-liposome-p53 resulted in 60.7% growth inhibition. Wild-type p53 expression and an increase in bax expression were observed following transfection with transferrin-liposome-p53, and 20.5% of the treated HOSM-1 cells were apoptotic. In vivo, the HOSM-1 tumor transplanted into nude mice grew to 5 to 6 mm in diameter. Following growth of the tumor to this size, transferrin-liposome p53 was locally applied to the peripheral tumor (day 0) and then applied once every 5 days for a total of six times. During the administration period, tumor growth did not occur, and the mean tumor volume on the last day of administration (day 25) was 10.0% of that in the saline control group. These results suggest that p53 gene therapy via cationic liposome modification with transferrin is an effective strategy for treatment of osteosarcoma. PMID- 15827337 TI - Targeted molecular therapy of anaplastic thyroid carcinoma with AEE788. AB - Anaplastic thyroid carcinoma (ATC) is one of the most aggressive human malignancies with a mean survival of only 6 months. The poor prognosis of patients with ATC reflects the current lack of curative therapeutic options and the need for development of novel therapeutic strategies. In this study, we report the results of a preclinical study of AEE788, a dual inhibitor of epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor (VEGFR) tyrosine kinases, against ATC. AEE788 was able to inhibit the proliferation and induce apoptosis of ATC cell lines in vitro. Administration of AEE788, alone and in combination with paclitaxel, to athymic nude mice bearing s.c. ATC xenografts inhibited the growth of ATC xenografts by 44% and 69%, respectively, compared with the control group. Furthermore, tumors from mice treated with AEE788, alone and in combination with paclitaxel, showed increase in apoptosis of tumor cells by approximately 6- and 8-fold, respectively, compared with the control group. The microvessel density within the ATC xenografts was decreased by >80% in the mice treated with AEE788 alone and in combination with paclitaxel compared with the control group. Lastly, immunofluorescence microscopy showed the inhibition of EGFR autophosphorylation on the tumor cells as well as the inhibition of VEGFR-2 autophosphorylation on tumor endothelium. Considering the fact that curative options seldom exist for patients with ATC, concurrent inhibition of EGFR and VEGFR tyrosine kinases seems to be a valid and promising anticancer strategy for these patients. PMID- 15827338 TI - Tumor penetration of gefitinib (Iressa), an epidermal growth factor receptor tyrosine kinase inhibitor. AB - The relative distribution of gefitinib-related material in nude mice bearing s.c. human tumor xenografts and in an orthotopic rat lung tumor model was investigated following oral administration (50 mg/kg) of [14C]-gefitinib. Selected tissue samples were monitored for radioactivity by liquid scintillation counting, whereas plasma and tumor extracts were assayed for gefitinib and its major metabolites (M523595 and M537194) by high-performance liquid chromatography with tandem mass spectrometric detection. Tissue distribution was also determined by whole body autoradiography. Gefitinib was extensively distributed into the tissues of tumor-bearing mice and unchanged gefitinib was shown to account for most of the tumor radioactivity. Concentrations of gefitinib in mouse s.c. tumor xenografts were similar to skin concentrations and substantially greater (up to 12-fold based on area under the concentration-time curve) than plasma. Concentrations of gefitinib-related material in an orthotopic rat lung tumor were similar to those in healthy lung tissue and were much higher than corresponding blood levels. Following treatment of breast cancer patients with oral gefitinib (Iressa) 250 mg/d for > or = 14 days, gefitinib concentrations (mean, 7.5 microg/g, 16.7 micromol/L) in breast tumor tissue were 42 times higher than plasma, confirming the preferential distribution of gefitinib from blood into tumor tissue in the clinical situation. These gefitinib tumor concentrations are considerably higher than those reportedly required in vitro to achieve complete inhibition of epidermal growth factor receptor autophosphorylation in both epidermal growth factor receptor mutant (0.2 micromol/L) and wild-type cells (2 micromol/L). PMID- 15827339 TI - Inhibition of the epidermal growth factor receptor increases expression of genes that stimulate inflammation, apoptosis, and cell attachment. AB - The epidermal growth factor receptor (EGFR) is overexpressed in several types of human cancer, and inhibition of EGFR function is a promising strategy for cancer therapy. We used cDNA microarrays to examine alterations in gene expression after treatment of carcinoma cells with PD153035, a specific and reversible inhibitor of EGFR function. When human cervical carcinoma cells were grown on a collagen substrate in three-dimensional organotypic culture, untreated cells expressed high levels of EGFR RNA and invaded the underlying collagen. Blocking EGFR function decreased DNA synthesis and inhibited invasion in a dose-dependent manner. Microarray analyses identified 312 genes that were significantly increased or decreased in expression after EGFR inhibition. Many could be classified into one of four functional groups including genes that (a) stimulate inflammation and innate immunity, (b) promote cell attachment, (c) enhance apoptosis, and (d) inhibit cell cycle progression. PD153035 induced a dose dependent activation of nuclear factor kappaB, a transcription factor that stimulates proinflammatory gene expression. Our results identify alterations in gene expression caused by EGFR inhibition and show that this response varies significantly in different cell lines. PMID- 15827340 TI - Targeted delivery to PEPT1-overexpressing cells: acidic, basic, and secondary floxuridine amino acid ester prodrugs. AB - Floxuridine is a clinically proven anticancer agent in the treatment of metastatic colon carcinomas and hepatic metastases. However, prodrug strategies may be necessary to improve its physiochemical properties and selectivity and to reduce undesirable toxicity effects. Previous studies with amino acid ester prodrugs of nucleoside drugs targeted to the PEPT1 transporter coupled with recent findings of the functional expression of the PEPT1 oligopeptide transporter in pancreatic adenocarcinoma cell lines suggest the potential of PEPT1 as therapeutic targets for cancer treatment. In this report, we show the feasibility of achieving enhanced transport and selective antiproliferative action of amino acid ester prodrugs of floxuridine in cell systems overexpressing PEPT1. All prodrugs exhibited affinity for PEPT1 (IC50, 1.1-2.3 mmol/L). However, only the prolyl and lysyl prodrugs exhibited enhanced uptake (2- to 8-fold) with HeLa/PEPT1 cells compared with HeLa cells, suggesting that the aspartyl prodrugs are PEPT1 inhibitors. The selective growth inhibition of Madine-Darby canine kidney (MDCK)/PEPT1 cells over MDCK cells by the prodrugs was consistent with the extent of their PEPT1-mediated transport. All ester prodrugs hydrolyzed to floxuridine fastest in Caco-2 cell and MDCK homogenates and slower in human plasma and were most chemically stable in pH 6.0 buffer. Prolyl and lysyl prodrugs were relatively less stable compared with aspartyl prodrugs in buffers and in cell homogenates. The results suggest that optimal design for targeted delivery would be possible by combining both stability and transport characteristics afforded by the promoiety. PMID- 15827341 TI - Variable expression of protein kinase C epsilon in human melanoma cells regulates sensitivity to TRAIL-induced apoptosis. AB - Protein kinase C (PKC) activation is believed to protect against apoptosis induced by death receptors. We have found however that the effect of activation of PKC on tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis of melanoma differs between cell lines. Pretreatment with phorbol 12 myristate 13-acetate (PMA) led to inhibition of apoptosis in the majority of the melanoma cell lines, but those with relatively low PKC epsilon expression were sensitized to TRAIL-induced apoptosis. Introduction of PKC epsilon into PKC epsilon-low cell lines reversed sensitization of the cells to TRAIL-induced apoptosis by PMA. In contrast, a dominant-negative form of PKC epsilon caused an increase in sensitivity. The changes in sensitivity to TRAIL-induced apoptosis were reflected in similar changes in conformation of Bax and its relocation from the cytosol to mitochondria. Similarly, there were concordant increases or decreases in mitochondrial release of second mitochondria-derived activator of caspase/DIABLO, activation of caspase-3, and processing of its substrates. Activation of PKC seemed to mediate its effects upstream of mitochondria but downstream of caspase-8 and Bid in that pretreatment with PMA did not cause significant changes in the expression levels of TRAIL death receptors, alterations in the levels of caspase-8 activation, or cleavage of Bid. PKC activated the anti-apoptotic extracellular signal-regulated kinase 1/2 pathway, but inhibitors of this pathway only partially reversed the protective effect of PKC against TRAIL-induced apoptosis. These results provide further insights into the variable responses of melanoma to TRAIL-induced apoptosis and may help define responsive phenotypes to treatment of melanoma with TRAIL. PMID- 15827342 TI - Raf kinase as a target for anticancer therapeutics. AB - The Ras-Raf-MEK-ERK (ERK) pathway is a logical therapeutic target because it represents a common downstream pathway for several key growth factor tyrosine kinase receptors which are often mutated or overexpressed in human cancers. Although considered mainly growth-promoting, in certain contexts, this pathway also seems to be apoptosis-suppressing. Several novel agents targeting this pathway have now been developed and are in clinical trials. One of the most interesting new agents is BAY 43-9006. Although initially developed as a Raf kinase inhibitor, it can also target several other important tyrosine kinases including VEGFR-2, Flt-3, and c-Kit, which contributes to its antiproliferative and antiangiogenic properties. To date, encouraging results have been seen with BAY 43-9006, particularly in renal cell cancers which are highly vascular tumors. This review will provide an overview of the ERK signaling pathway in normal and neoplastic tissue, with a specific focus on novel therapies targeting the ERK pathway at the level of Raf kinase. PMID- 15827343 TI - Proteasome inhibitor therapy in multiple myeloma. AB - Multiple myeloma remains incurable despite available therapies, and novel therapies that target both tumor cell and bone marrow microenvironment are urgently needed. Preclinical in vitro and in vivo studies show remarkable anti multiple myeloma activity of the proteasome inhibitor bortezomib/PS-341 even in multiple myeloma cells refractory to multiple prior therapies, including dexamethasone, melphalan, and thalidomide. Based on these findings, the U.S. Food and Drug Administration recently approved the first proteasome inhibitor bortezomib (Velcade), formerly known as PS-341, for the treatment of relapsed/refractory multiple myeloma. Bortezomib therapy has set an outstanding example of translational research in the field of oncology. Genomics and proteomic studies further provide rationale for combining bortezomib with conventional and novel agents to inhibit multiple myeloma growth, overcome drug resistance, reduce attendant toxicity, and improve patient outcome in multiple myeloma. PMID- 15827344 TI - High ambient glucose is effect neutral on cell death and proliferation in human proximal tubular epithelial cells. AB - In vitro models of diabetic nephropathy that assess the role of hyperglycemia on proximal tubular cell turnover commonly compare cells in a high-glucose medium (25 or 30 mM) with a low-glucose medium (5 to 6.1 mM). Any cellular growth changes observed are usually attributed to the effect of high glucose. We hypothesize that in such experiments, glucose concentrations in the low-glucose medium may decline during the course of the experiments to levels that inhibit cell growth leading to the comparative conclusion that high glucose induces hyperplasia and/or hypertrophy. In this study, primary cultures of human proximal tubular epithelial cells (PTEC) and immortalized HK-2 cells were exposed to low (5 mM) or high (17, 30, or 47 mM) glucose for up to 6 days (PTEC) and 48 h (HK 2). When culture media were not replenished, low glucose induced a significant increase in necrosis and release of lactate dehydrogenase and a decrease in proliferation, metabolic activity, and protein content without any changes in apoptosis. High-glucose media failed to induce any of these changes. Glucose was undetectable in the low-glucose culture medium after 72 h. No significant differences were observed between any of the treatment groups when culture media were replenished daily. We conclude that regular replenishment of culture media is necessary to prevent the emergence of artifactual and misleading differences between high- and low-glucose groups. The current knowledge of the pathophysiology of high glucose based on cell culture systems may need to be reevaluated. PMID- 15827345 TI - Differential effects of salt on renal hemodynamics and potential pressure transmission in stroke-prone and stroke-resistant spontaneously hypertensive rats. AB - Salt-supplemented stroke-prone spontaneously hypertensive rats (SHRsp) develop more severe hypertension-induced renal damage (HIRD) compared with their progenitor SHR. The present studies were performed to examine whether in addition to increasing the severity of hypertension salt also enhanced the transmission of such hypertension to the renal vascular bed in the SHRsp. "Step" and "dynamic" renal blood flow (RBF) autoregulation (AR) were examined in approximately 12-wk old SHR and SHRsp after 3-5 days of an 8% NaCl diet. During step AR under anesthesia (n = 8-11), RBF was significantly higher in the SHRsp at all perfusion pressures (P < 0.01), but AR capacity was not different. Similarly, in separate conscious chronically instrumented rats (n = 8 each), both blood pressure (BP) and RBF were modestly but significantly higher at baseline before salt in the SHRsp (P < 0.05). However, transfer function analysis did not show significant differences in the admittance gain parameters. However, after 3-5 days of salt, although average BP was not significantly altered in either strain, RBF increased further in the SHRsp and there was a significantly greater transfer of BP into RBF power in the SHRsp. This was reflected in the significantly higher admittance gain parameters at most frequencies including the heartbeat frequency (P < 0.05 maximum). These differential hemodynamic effects of salt have the potential to enhance BP transmission to the renal vascular bed and also contribute to the more severe HIRD observed in the salt-supplemented SHRsp. PMID- 15827346 TI - Kidney expression of glutathione peroxidase-1 is not protective against streptozotocin-induced diabetic nephropathy. AB - In many diseases, including progressive renal disorders, tissue injury and pathological intracellular signaling events are dependent on oxidative stress. Glutathione peroxidase-1 (Gpx1) is an antioxidant enzyme that is highly expressed in the kidney and removes peroxides and peroxynitrite that can cause renal damage. Therefore, we examined whether this abundant renal antioxidant enzyme limits renal damage during the development of type 1 diabetic nephropathy. Wild type (Gpx1+/+) and deficient (Gpx1-/-) mice were made diabetic by intraperitoneal injection of streptozotocin (100 mg/kg) on 2 consecutive days. Diabetic Gpx1+/+ and -/- mice with equivalent blood glucose levels (23 +/- 4 mM) were selected and examined after 4 mo of diabetes. Compared with normal mice, diabetic Gpx1+/+ and /- mice had a two- to threefold increase in urine albumin excretion at 2 and 4 mo of diabetes. At 4 mo, diabetic Gpx1+/+ and -/- mice had equivalent levels of oxidative renal injury (increased kidney reactive oxygen species, kidney lipid peroxidation, urine isoprostanes, kidney deposition of advanced glycoxidation, and nitrosylation end products) and a similar degree of glomerular damage (hypertrophy, hypercellularity, sclerosis), tubular injury (apoptosis and vimentin expression), and renal fibrosis (myofibroblasts, collagen, TGF-beta excretion). A lack of Gpx1 was not compensated for by increased levels of catalase or other Gpx isoforms in diabetic kidneys. Contrary to expectations, this study showed that the high level of Gpx1 expressed in the kidney is not protective against the development of renal oxidative stress and nephropathy in a model of type 1 diabetes. PMID- 15827347 TI - Urodynamic properties and neurotransmitter dependence of urinary bladder contractility in the BK channel deletion model of overactive bladder. AB - Overactive bladder and incontinence are major medical issues, which lack effective therapy. Previously, we showed (Meredith AL, Thornloe KS, Werner ME, Nelson MT, and Aldrich RW. J Biol Chem 279: 36746-36752, 2004) that the gene mSlo1 encodes large-conductance Ca2+-activated K+ (BK) channels of urinary bladder smooth muscle (UBSM) and that ablation of mSlo1 leads to enhanced myogenic and nerve-mediated contractility and increased urination frequency. Here, we examine the in vivo urodynamic consequences and neurotransmitter dependence in the absence of the BK channel. The sensitivity of contractility to nerve stimulation was greatly enhanced in UBSM strips from Slo-/- mice. The stimulation frequency required to obtain a 50% maximal contraction was 8.3 +/- 0.9 and 19.1 +/- 1.8 Hz in Slo-/- and Slo+/+ mice, respectively. This enhancement is at least partially due to alterations in UBSM excitability, as muscarinic induced Slo-/- contractility is elevated in the absence of neuronal activity. Muscarinic-induced Slo-/- contractility was mimicked by blocking BK channels with iberiotoxin (IBTX) in Slo+/+ strips, whereas IBTX had no effect on Slo-/- strips. IBTX also enhanced purinergic contractions of Slo+/+ UBSM but was without effect on purinergic contractions of Slo-/- strips. In vivo bladder pressure and urine output measurements (cystometry) were performed on conscious, freely moving mice. Slo-/- mice exhibited increased bladder pressures, pronounced pressure oscillations, and urine dripping. Our results indicate that the BK channel in UBSM has a very significant role in urinary function and dysfunction and as such likely represents an important therapeutic target. PMID- 15827348 TI - Enhanced expression of EGF receptor in a model of salt-sensitive hypertension. AB - Chronic kidney disease in the Dahl/Rapp salt-sensitive (S) rat is related to an arteriolopathic process that occurs following the onset of hypertension and involves vascular smooth muscle cell (VSMC) hyperplasia and luminal constriction. Because previous studies have shown that activation of the epidermal growth factor receptor (EGFR) produces a mitogenic stimulus in VSMC and the EGFR participates integrally in the vasoconstrictor responses of renal arterioles, the present study analyzed the expression of EGFR in these animals. Compared with Sprague-Dawley (SD) rats, renal cortical expression of EGFR was increased in both prehypertensive and hypertensive S rats. Immunohistochemistry using a polyclonal antibody to EGFR demonstrated that EGFR expression was prominent in the renal vasculature, particularly in the media of afferent and efferent arterioles and the aorta of S rats. When examined, primary cultures of VSMC from S rats showed increased expression of EGFR, compared with VSMC from SD and Dahl/Rapp salt resistant rats. Following addition of EGF, autophosphorylation of the EGFR was enhanced in cells from S rats, as was the downstream signaling events that included activation of p42/44 MAPK and Akt pathways. Thus in vivo and in vitro studies demonstrated augmented expression and functional activity of the EGFR in S rats. PMID- 15827349 TI - Outcome from a prospective, longitudinal study of prenatal cocaine use: preschool development at 3 years of age. AB - OBJECTIVE: To determine the effects of prenatal cocaine exposure on child development. METHODS: This prospective, longitudinal study recruited 154 pregnant cocaine users who were matched on race, parity, socioeconomic status, and perinatal risk to 154 noncocaine users. Drug use status was determined by maternal history and urine screening. At 3 years of age, the child subjects were assessed by an evaluator blinded to maternal drug use history. During a home visit at age 3, caregiver, family, and home assessments were administered. RESULTS: Structural equation modeling showed a direct effect of the amount of prenatal cocaine exposure on the adjusted birth head circumference which in turn directly affected preschool development. CONCLUSIONS: We could not demonstrate a direct effect of prenatal cocaine exposure on preschool development, a result that is consistent with that of earlier work and now extending findings to age 3. However, cocaine continued to exert an indirect effect on development through its direct effect on the head circumference at birth. PMID- 15827350 TI - Interactions between maternal characteristics and neonatal behavior in the prediction of parenting stress and perception of infant temperament. AB - BACKGROUND: Prenatal cocaine exposure is a marker of developmental risk. Social environmental risk factors may include maternal stress and maternal perceptions of difficult infant temperament. OBJECTIVES: To examine factors that may predict or moderate maternal ratings of parenting stress and difficult temperament in cocaine-exposed (CE) infants. METHOD: Neonatal behavior, infant temperament, parenting stress, and maternal psychopathology were measured in a large sample of infant-mother dyads with prenatal CE and a nonexposed comparison sample. Participants were drawn from an existing longitudinal data set (Maternal Lifestyle Study). RESULT: Relations between neonatal behavior and infant temperament ratings were moderated by mothers' ratings of parenting stress. Relations between neonatal cry and parenting stress were moderated by maternal psychopathology ratings. Results were unrelated to drug exposure history. CONCLUSIONS: For mothers of at risk infants (with or without prenatal CE), psychological distress affects the degree to which infant behavioral characteristics are experienced as stressful or difficult. Implications for treatment and outcome are discussed. PMID- 15827352 TI - Supportive relationships and sexual risk behavior in adolescence: an ecological transactional approach. AB - OBJECTIVE: To examine the longitudinal associations between supportive relationships with friends and parents and sexual risk behavior in adolescence based on an ecological-transactional perspective. METHODS: Analyses were conducted on 2,652 sexually active adolescents from the first two waves of the National Longitudinal Study of Adolescent Health (Add Health). RESULTS: African American adolescents had lower risk for sexual risk behavior. Supportive friendships and parent connectedness interacted in predicting decreased likelihood of sexual risk behavior. Mother-child communication about sex contributed to decreased likelihood of sexual risk only for girls. There were also small reciprocal effects of sexual risk behavior on decreased relationship quality over time. CONCLUSION: To better understand the parents' role in adolescent sexual risk behavior, multiple facets of parenting, the social contexts of parenting and adolescents' peers, and the effects of adolescents' behavior on these relationships should be taken into consideration. PMID- 15827351 TI - Aggression at age 5 as a function of prenatal exposure to cocaine, gender, and environmental risk. AB - OBJECTIVE: To examine childhood aggression at age 5 in a multiple risk model that includes cocaine exposure, environmental risk, and gender as predictors. METHODS: Aggression was assessed in 206 children by using multiple methods including teacher report, parent report, child's response to hypothetical provocations, and child's observed behavior. Also examined was a composite score that reflected high aggression across contexts. RESULTS: Multiple regression analyses indicated that a significant amount of variance in each of the aggression measures and the composite was explained by the predictors. The variables that were independently related differed depending on the outcome. Cocaine exposure, gender, and environmental risk were all related to the composite aggression score. CONCLUSIONS: Cocaine exposure, being male, and a high-risk environment were all predictive of aggressive behavior at 5 years. It is this group of exposed boys at high environmental risk that is most likely to show continued aggression over time. PMID- 15827353 TI - Dissecting the axoneme interactome: the mammalian orthologue of Chlamydomonas PF6 interacts with sperm-associated antigen 6, the mammalian orthologue of Chlamydomonas PF16. AB - The axoneme central apparatus is thought to control flagellar/ciliary waveform and maintain the structural integrity of the axoneme, but proteins involved in these processes have not been fully elucidated. Moreover the network of interactions among them that allows these events to take place in a compact space has not been defined. PF6, a component of the Chlamydomonas central apparatus, is localized to the 1a projection of the C1 microtubule. Mutations in the Chlamydomonas PF6 gene result in flagellar paralysis. We characterized human and murine orthologues of PF6. The murine Pf6 gene is expressed in a pattern consistent with a role in flagella and cilia, and the PF6 protein is indeed localized to the central apparatus of the sperm flagellar axoneme. We discovered that a portion of PF6 associates with the mammalian orthologue of Chlamydomonas PF16 (sperm-associated antigen 6 (SPAG6)), another central apparatus protein that is localized to the C1 microtubule in algae. A fragment of PF6 corresponding to the PF6 domain that interacts with SPAG6 in yeast two-hybrid assays and colocalizes with SPAG6 in transfected cells was missing from epididymal sperm of SPAG6-deficient mice. SPAG6 binds to the mammalian orthologue of PF20, which in Chlamydomonas is located in bridges connecting the C2 and C1 microtubules. Thus, PF6, SPAG6, and PF20 form a newly identified network that links together components of the axoneme central apparatus and presumably participates in its dynamic regulation of ciliary and flagellar beat. PMID- 15827354 TI - The effectiveness of reconstruction of the anterior cruciate ligament using the novel knot/press-fit technique: a cadaveric study. AB - BACKGROUND: A variety of fixation devices are used for anterior cruciate ligament reconstruction with hamstring tendon grafts. These devices increase costs and can present artifacts in magnetic resonance imaging as well as complications in revision surgery. Therefore, a novel knot/press-fit technique that requires no implantable devices has been introduced. NULL HYPOTHESIS: The knot/press-fit technique restores knee kinematics as well as the more commonly used EndoButton CL fixation and has similar biomechanical properties as other devices published in the literature. STUDY DESIGN: Controlled laboratory study. METHODS: Eight fresh-frozen cadaveric knees (52 +/- 7 years) were tested using a robotic/universal force-moment sensor testing system. The knee kinematics of the intact, anterior cruciate ligament-deficient, EndoButton-reconstructed, and knot/press-fit-reconstructed knee in response to both a 134-N anterior tibial load and a combined rotatory load at multiple knee flexion angles was determined. Differences between the 4 knee states were evaluated with a 2-factor repeated measures analysis of variance (P < .05). To determine the stiffness and strength of the knot/press-fit fixation, the femur-graft-tibia complex was tested in uniaxial tension. RESULTS: In response to an anterior tibial load, the anterior tibial translation for the knot/press-fit reconstruction was found to be not significantly different from that of the intact anterior cruciate ligament as well as that of the EndoButton reconstruction (P > .05). In response to a combined rotatory load, neither reconstruction procedure could effectively reduce the coupled anterior tibial translation to that of the intact knee, and no significant difference between the 2 reconstructions could be detected (P > .05). The stiffness of the knot/press-fit complex was found to be 37.8 +/- 9.6 N/mm, and the load at failure was 540 +/- 97.7 N, which is equal to other devices published in the literature. CLINICAL RELEVANCE: The experiment suggests that the knot/press-fit technique may be a reliable alternative for the femoral fixation of hamstring tendon grafts. PMID- 15827355 TI - Location of the glenoid defect in shoulders with recurrent anterior dislocation. AB - BACKGROUND: In patients with recurrent anterior dislocations of the shoulder, it is well known that the glenoid rim is often deficient (8%-95%). However, little is known regarding the precise location of the bony defect of the glenoid. HYPOTHESIS: The bony defect is anterior rather than anteroinferior to the glenoid. STUDY DESIGN: Cohort study (symptom prevalence); Level of evidence, 2. METHODS: The authors studied 3-dimensional images of the glenoid reconstructed from computed tomography to determine the location of the glenoid defect in 123 shoulders of 123 patients with recurrent anterior dislocations of the shoulder. They measured the location, extent, and orientation of the defect based on the clock face of the glenoid. RESULTS: The defects were located between 12:08 and 6:32, with the range between 2:30 and 4:20 being the most frequent. The extent of the glenoid defect was 106.7 degrees +/- 27.1 degrees (mean +/- standard deviation). The mean orientation of the defect was pointing toward 3:01 on the clock face of the glenoid, at a mean angle of 90.5 degrees +/- 10.4 degrees from the 12-o'clock direction. CONCLUSION: The glenoid defect is located almost anterior to the glenoid. PMID- 15827356 TI - Effect of drain placed in the donor site in the early postoperative period after arthroscopically assisted anterior cruciate ligament reconstruction with quadrupled hamstring tendons. AB - BACKGROUND: No scientific study has addressed the effects of the routine use of postoperative drains in the donor site after arthroscopic anterior cruciate ligament reconstruction using hamstring tendons. HYPOTHESIS: Patients who have drains placed in their donor sites have less lower extremity edema, more motion, and more comfort in the early postoperative period. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: Thirty-four patients undergoing arthroscopically assisted quadruple hamstring tendon anterior cruciate ligament reconstruction were randomly assigned to either receive a drain for 24 hours placed at the donor site (17 patients) or have no drain at the donor site (17 patients). Data were collected on postoperative days 1, 3, 5, and 7 regarding knee flexion, knee extension, thigh circumference, leg circumference, and visual analog scale (measuring pain). RESULTS: In the study group versus the control group, there were increased knee flexion measurements on postoperative day 7 (84 degrees vs 69 degrees , P < .05); smaller thigh circumference measurements on postoperative day 7 (36.8 vs 40.1 cm, P < .05); smaller leg circumference measurements on postoperative days 3 (33.2 vs 36.4 cm, P < .05), 5 (32.7 vs 36.0 cm, P < .05), and 7 (31.8 vs 35.7 cm, P < .001); and a lower visual analog scale score on postoperative days 3 (40.38 vs 57.50, P = .001), 5 (38.46 vs 60.35, P = .001), and 7 (38.07 vs 61.43, P = .001). CONCLUSION: Drain placement at the donor site after anterior cruciate ligament reconstruction gives more comfort to the patient and provides better flexion within the first week. PMID- 15827357 TI - Percutaneous versus open repair of the ruptured Achilles tendon: a comparative study. AB - BACKGROUND: Controversy regarding the optimal treatment of the fresh total Achilles tendon rupture remains. PURPOSE: To compare the results of percutaneous and open Achilles tendon repair. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: The results of 132 consecutive patients with acute complete Achilles tendon rupture who were operated on exclusively with modified percutaneous repair under local anesthesia from 1991 to 1997 and followed up for at least 2 years were compared to the results of 105 consecutive patients who were operated on exclusively with open repair under general or spinal anesthesia in the same period. RESULTS: There were significantly fewer major complications in the group of percutaneous repairs in comparison with the group of open repairs (4.5% vs 12.4%; P = .03), particularly necrosis (0% vs 5.6%; P = .019), and a lower total number of complications (9.7% vs 21%; P = .013). There were slightly more reruptures (3.7% vs 2.8%; P = .680) and sural nerve disturbances (4.5% vs 2.8%; P = .487) in the group of percutaneous repairs, with no statistically significant difference. Functional assessment using the American Orthopaedic Foot and Ankle Society scale and the Holz score showed no statistically significant difference. CONCLUSION: The results of the study support the choice of (modified) percutaneous suturing under local anesthesia as the method that brings comparable functional results to open repair, with a significantly lower rate of complications. PMID- 15827358 TI - Early inflammatory reaction after rotator cuff repair with a porcine small intestine submucosal implant: a report of 4 cases. AB - BACKGROUND: Porcine small intestine submucosal grafts have been successful in enhancing soft tissue repair, as demonstrated by animal studies. Currently, there are no reports of the use of such implants in human rotator cuff repair. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Over a 6-month period, 25 patients underwent rotator cuff repair by one surgeon using the Restore Orthobiologic Implant to augment the repaired tendon or fill a defect. RESULTS: Four of 25 patients experienced an overt inflammatory reaction at a mean of 13 days postoperatively. All patients underwent open irrigation and debridement of the rotator cuff and porcine small intestine submucosal implant. CONCLUSION: Porcine small intestine submucosal implants should be used in rotator cuff surgery with the awareness that a non-specific inflammatory reaction can occur in the early postoperative period. This inflammatory reaction may cause breakdown of the repair. Further studies are needed to further characterize the reaction and determine which patients are susceptible. PMID- 15827359 TI - Instruction of jump-landing technique using videotape feedback: altering lower extremity motion patterns. AB - BACKGROUND: Anterior cruciate ligament injury prevention programs have used videotapes of jump-landing technique as a key instructional component to improve landing performance. HYPOTHESIS: All videotape feedback model groups will increase knee flexion angles at initial contact and overall knee flexion motion and decrease peak vertical ground reaction forces and peak proximal anterior tibial shear forces to a greater extent than will a nonfeedback group. The secondary hypothesis is that the videotape feedback using the combination of the expert and self models will create the greatest change in each variable. STUDY DESIGN: Controlled laboratory study. METHODS: Knee kinematics and kinetics of college-aged recreational athletes randomly placed in 3 different videotape feedback model groups (expert only, self only, combination of expert and self) and a nonfeedback group were collected while participants performed a basketball jump-landing task on 3 testing occasions. RESULTS: All feedback groups significantly increased knee angular displacement flexion angles [F(6,70) = 8.03, P = .001] and decreased peak vertical ground reaction forces [F(6,78) = 2.68, P = .021] during performance and retention tests. The self and combination groups significantly increased knee angular displacement flexion angles more than the control group did; the expert model group did not change significantly more than the control group did. All feedback groups and the nonfeedback group significantly reduced peak vertical forces across performance and retention tests. There were no statistically significant changes in knee flexion angle at initial ground contact (P = .111) and peak proximal anterior tibial shear forces (P = .509) for both testing sessions for each group. CONCLUSION: The use of self or combination videotape feedback is most useful for increasing knee angular displacement flexion angles and reducing peak vertical forces during landing. CLINICAL RELEVANCE: The use of self or combination modeling is more effective than is expert-only modeling for the implementation of instructional programs aimed at reducing the risk of jump-landing anterior cruciate ligament injuries. PMID- 15827360 TI - The posterolateral corner of the knee: repair versus reconstruction. AB - BACKGROUND: Injuries to the posterolateral corner of the knee are disabling. Despite improved understanding of this group of tendons and ligaments, the best surgical treatment of an unstable corner is not clear. HYPOTHESIS: Surgical repair of acute tears of the posterolateral corner has outcomes that are as good as those from reconstruction combined with an early motion rehabilitation protocol. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Sixty-three patients with 64 posterolateral corner tears were included in this study, with 39 posterolateral corner repairs and 25 reconstructions using the modified 2-tailed technique. Patients were evaluated with clinical and KT-2000 arthrometer examinations, as well as with Lysholm, International Knee Documentation Committee, and Short Form-36 scores. RESULTS: Fifty-six patients with 57 corner tears had minimum clinical follow-up of 24 months (range, 24-59 months). Acute primary repairs were performed on 35 patients, with 22 successful outcomes and 13 (37%) failures. Primary reconstructions were performed on 22 patients, with 20 successful outcomes and 2 (9%) failures. The difference in stability on clinical examination between repairs and reconstructions was significant (P < .05). Fourteen of 15 patients with failures of the primary posterolateral corner repair or reconstruction underwent successful revision reconstruction. The final patient with failure of the primary repair elected not to have a revision reconstruction. The final mean Lysholm knee score for both repair and reconstruction patients was 88.7 (range, 53-100). Final International Knee Documentation Committee objective scores yielded 14 (26%) normal, 28 (52%) near-normal, 9 (17%) abnormal, and 3 (6%) severely abnormal knees; the mean score was 60 at the most recent clinical evaluation. CONCLUSION: Results with repair followed by early motion rehabilitation have been significantly inferior when compared with results from reconstruction using the modified 2-tailed technique. The authors now use reconstruction rather than repair in the majority of patients who sustain posterolateral corner tears after high-energy injuries. PMID- 15827361 TI - Variations in mortise anatomy. AB - BACKGROUND: Variations in ankle mortise anatomy may be a predisposing factor to ankle instability. HYPOTHESIS: A posteriorly positioned fibula associated with ankle instability may not be a true pathologic entity but rather the result of measuring off an internally rotated talus. STUDY DESIGN: Cohort study (diagnosis); Level of evidence, 2. METHODS: The authors reviewed 60 ankle computed tomography scans performed on patients from their institution for reasons unrelated to ankle instability. They also reviewed ankle computed tomography scans on 21 patients surgically treated for clinical ankle instability. The position of the fibula in relation to the talar articular surface was calculated and expressed as the axial malleolar index, as described by Scranton et al. They also calculated the intermalleolar index, a new method that references the medial malleolus, not the talus. RESULTS: Using the method of Scranton et al, the axial malleolar index in the control and instability patients was similar to values previously described, and there was a significant difference between control and instability patients (P < .01). However, using the intermalleolar index method referencing the medial malleolus, there was not a significant difference between control and instability patients (P = .43). CONCLUSION: The new method of referencing the medial malleolus assesses fibular position independent of talar rotation. The data, when referencing the medial malleolus, do not show significant variation in fibular position in patients with and without ankle instability. PMID- 15827362 TI - Groin pain in footballers: the association between preseason clinical and pubic bone magnetic resonance imaging findings and athlete outcome. AB - BACKGROUND: Groin pain and tenderness are common in athletes from a variety of codes of football, but little attention has been directed to the influence of magnetic resonance imaging and such clinical findings on athlete participation. HYPOTHESIS: Preseason groin pain, tenderness, and magnetic resonance imaging findings such as pubic bone marrow edema are associated with restricted training capacity and missed games. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Fifty-two Australian footballers in the national competition were recruited. Preseason groin pain and focal tenderness were recorded, and magnetic resonance imaging of the groin was performed within 1 week of examination. Training restriction and games missed owing to groin pain were documented during the subsequent season. RESULTS: Magnetic resonance imaging showed pubic bone marrow edema in 19 of 52 (37%) footballers and linear parasymphyseal T2 hyperintensity in 16 of 52 (31%) footballers. Groin pain restricted training during the season in 22 of 52 (42%) footballers, and 9 of 52 (17%) footballers missed at least 1 game. Preseason pain (P = .0004), pubic bone tenderness (P = .02), and linear parasymphyseal T2 hyperintensity (P = .01) were associated with restricted training capacity during the subsequent season. Preseason groin pain (P = .03) was associated with missed games, but magnetic resonance imaging findings were not. CONCLUSION: Preseason pubic bone marrow edema, groin pain, and linear parasymphyseal T2 hyperintensity were associated with training restriction, but only preseason groin pain was associated with missed games. PMID- 15827363 TI - Snowboarder's talus fracture: treatment outcome of 20 cases after 3.5 years. AB - BACKGROUND: Fracture of the lateral process of the talus is a typical snowboarding injury. Basic data are limited, particularly with respect to treatment and outcome. HYPOTHESIS: As the axial-loaded dorsiflexed foot becomes externally rotated and/or everted, fracture of the lateral process of the talus occurs. Primary surgical treatment may improve the outcome of this injury, reducing the risk of secondary subtalar joint osteoarthritis. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: We recorded details of the treatment and evaluation of 20 patients (8 female and 12 male; age at trauma, 29 years [range, 17-48 years]) who sustained a lateral process of the talus fracture while snowboarding. The injury pathomechanism was documented. The patients were treated either nonsurgically or surgically based on a fracture-type treatment algorithm. The evaluation at most recent follow-up (mean, 42 months [range, 26-53 months]) included clinical and functional examination, follow-up of sport activity, and radiological assessment (radiograph, computed tomography scan). RESULTS: The injury mechanism included axial impact (100%), dorsiflexion (95%), external rotation (80%), and eversion (45%). Using the American Orthopaedic Foot and Ankle Society hindfoot score, the patients obtained a mean of 93 points; the surgically treated group (n = 14) scored higher (97 points) than did the nonoperative group (n = 6; 85 points) (P < .05). Degenerative disease of the subtalar joint was found in 3 patients (15%; operative, 1 patient; nonoperative, 2 patients). All but 4 (20%, all after nonsurgical treatment) patients reached the same sport activity level as before injury. CONCLUSION: The snowboarding-related lateral process of the talus fracture represents a complex hindfoot injury. In type II fractures, primary surgical treatment has led to achieving better outcomes, reducing sequelae, and allowing patients to regain the same sports activity level as before injury. PMID- 15827364 TI - Hydration status in college football players during consecutive days of twice-a day preseason practices. AB - BACKGROUND: Football players lose 3.5 to 5 kg of body weight during preseason practices because of heavy sweating. This fluid may be difficult to replace when practices occur 2 times per day on consecutive days. HYPOTHESIS: Football players are hypohydrated during twice-a-day preseason training in a hot, humid environment. STUDY DESIGN: Descriptive laboratory study. METHODS: In 10 college football players, body weight was measured, and blood and urine samples were obtained before and after practices on days 2 through 8 of preseason training. Baseline samples were obtained when subjects were euhydrated. Blood samples were used to calculate plasma volume changes. Urine samples were analyzed for specific gravity, sodium, and potassium. Sweat rate was calculated. Core temperature was monitored during half- and full-padded practices. RESULTS: Mean wet bulb temperatures were 23.3 degrees C during morning practices and 23.7 degrees C during afternoon practices. Plasma volume was below baseline on day 2 and expanded by day 6. Urine specific gravity was higher than baseline for 12 of 20 measurements over the 8 days. It was 1.0175 +/- 0.006 at baseline but subsequently ranged from 1.0214 +/- 0.007 to 1.0321 +/- 0.004. Mean daily urine sodium dropped from baseline to day 2 (194 +/- 43 vs 43 +/- 38 mmol x L-1), remaining lower on days 3, 4, and 6 (40 +/- 39, 39 +/- 39, and 68 +/- 40 mmol x L 1, respectively). Urine potassium was lower on days 6 and 8 compared with baseline and day 3. Body weight was below baseline before and after both daily practices. Core temperature was higher in full pads; sweat rate and body weight loss were not different between half and full pads. CONCLUSION: Body weight, plasma volume, urine specific gravity, and urine sodium indicate that football players become dehydrated by day 2 of preseason training. Urine sodium increased to near baseline by day 8; urine specific gravity was elevated. CLINICAL RELEVANCE: Football players struggle to maintain euhydration during preseason twice-a-day sessions. PMID- 15827365 TI - Topical glyceryl trinitrate application in the treatment of chronic supraspinatus tendinopathy: a randomized, double-blinded, placebo-controlled clinical trial. AB - BACKGROUND: Topical glyceryl trinitrate therapy has previously demonstrated short term pain reduction in patients with supraspinatus tendinopathy. HYPOTHESIS: Topical glyceryl trinitrate improves outcome measures in patients with supraspinatus tendinopathy. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: Fifty-three patients (57 shoulders) were recruited, and the authors completed a prospective, randomized, double-blinded, placebo-controlled clinical trial of continuous topical glyceryl trinitrate treatment (1.25 mg/24-h glyceryl trinitrate). RESULTS: The glyceryl trinitrate group had significantly reduced shoulder pain with activity, at night, and at rest at week 24 (P = .03); reduced internal rotation impingement at week 24 (P = .02); increased range of motion in abduction and internal rotation at week 24 (P = .04); and increased force at weeks 12 and 24 with supraspinatus muscle testing (P = .001), external rotation (P = .04), internal rotation (P = .01), adduction (P = .04), and subscapularis pushoff (P = .01). Forty-six percent of patients on glyceryl trinitrate patches were asymptomatic with activities of daily living at 6 months compared with 24% of patients with tendon rehabilitation alone (P = .007, chi(2) analysis). Mean effect size for all outcome measures was 0.26. DISCUSSION: Topical glyceryl trinitrate treatment significantly improved pain scores, range of motion, internal rotation impingement, muscular force, and patient outcomes in patients with supraspinatus tendinopathy. Topical glyceryl trinitrate should be included as part of nonsurgical management of chronic tendinopathies. PMID- 15827366 TI - Sport-dependent variations in arm position during single-limb landing influence knee loading: implications for anterior cruciate ligament injury. AB - BACKGROUND: Increased valgus loading at the knee has been previously identified as a possible risk factor for noncontact anterior cruciate ligament injuries, which are common in sports. Arm position variation may affect risk of injury by altering valgus knee loading. HYPOTHESIS: Sport-dependent variations in arm position increase valgus loading of the knee during run-to-cut maneuvers. STUDY DESIGN: Controlled laboratory study. METHODS: Eleven subjects performed a sidestep cutting maneuver, first with no arm constraints and then with 3 sports related arm positions in random order (holding a lacrosse stick, holding a football on the plant side, and holding a football on the cut side). The analysis focused on the knee valgus moment relative to the arm positions during the landing phase of the activity. RESULTS: Arm position significantly influenced the valgus moment with an increase in the lacrosse trials and in the plant-side football trials but not in the cut-side football trials (alpha = .05). CONCLUSIONS: Constraining the plant-side arm results in increased valgus loading at the knee during run-to-cut maneuvers, which suggests the possibility of greater risk of anterior cruciate ligament injury during these conditions. CLINICAL RELEVANCE: These results suggest that training methods that consider arm position as a risk factor could help reduce the risk of anterior cruciate ligament noncontact injury. PMID- 15827367 TI - Correlation of Short Form-36 and disability status with outcomes of arthroscopic acetabular labral debridement. AB - BACKGROUND: Arthroscopic debridement is the standard of care for the treatment of acetabular labral tears. The Short Form-36 has not been used to measure hip arthroscopy outcomes, and the impact of disability status on hip arthroscopy outcomes has not been reported. HYPOTHESIS: Short Form-36 subscale scores will demonstrate good correlation with the modified Harris hip score, but patients undergoing disability evaluation will have significantly worse outcome scores. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The records of active duty soldiers who underwent hip arthroscopy at the authors' institution were retrospectively reviewed. Forty consecutive patients who underwent hip arthroscopy for the primary indication of labral tear formed the basis of the study group. Patients completed the modified Harris hip score, the Short Form-36 general health survey, and a subjective overall satisfaction questionnaire. RESULTS: Thirty-three patients, with a mean age of 34.6 years, were available for follow-up at a mean of 25.7 months postoperatively. Fourteen (43%) patients were undergoing medical evaluation boards (military equivalent of workers' compensation or disability claim). Pearson correlation coefficients for comparing the Short Form-36 Bodily Pain, Physical Function, and Physical Component subscale scores to the modified Harris hip score were 0.73, 0.71, and 0.85, respectively (P < .001). The mean modified Harris hip score was significantly lower in patients on disability status than in those who were not (92.4 vs 61.1; P < .0001). The Short Form-36 subscale scores were significantly lower in disability patients (P < .02). Patient-reported satisfaction rates (70% overall) were 50% for those undergoing disability evaluations and 84% for those who were not (P < .04). There was no significant difference in outcomes based on patient age, surgically proven chondromalacia, or gender for military evaluation board status. CONCLUSION: The Short Form-36 demonstrated good correlation with the modified Harris hip score for measuring outcomes after arthroscopic partial limbectomy. Arthroscopic debridement yielded a high percentage of good results when patients undergoing disability evaluations were excluded. Disability status may be a negative predictor of success after hip arthroscopy. PMID- 15827368 TI - Bioterrorism, Glanders and melioidosis. PMID- 15827369 TI - Survey of the contamination of foodstuffs of animal origin by Shiga toxin producing Escherichia coli serotype O157:H7 in Belgium from 1999 to 2003. AB - A survey of the prevalence of Shiga toxin-producing Escherichia coli (STEC) of O157 serotype in foodstuffs of animal origin (beef, veal, pork, chicken, fish) from 1999 to 2003 in Belgium was performed. STEC strains were only isolated from beef with a prevalence of 0.73%. This percentage is low in comparison with the prevalence in other countries. Among the 76 isolated STEC O157 strains, 75% belonged to the serotype O157:H7 and 25% to the serotype O157 non H7. Moreover, the most frequent pathotype was eae stx2 ehxA (74%). PMID- 15827370 TI - Communicable disease control in a migrant seasonal workers population: a case study in Norway. AB - Reliable data on the health status of migrant seasonal workers in Europe is scarce. Access to public health care for this population depends on national regulations, and their legal status in host countries. In this manuscript we describe a case study of a salmonellosis outbreak that occurred in Norway, and highlight the difficulties encountered in applying control measures in a population of seasonal migrant farm workers. Surveillance and control of infectious diseases need to be supported by legislation which makes implementation of control measures possible. Efforts have been made to improve the rights for migrants in Europe with regard to healthcare, but seasonal migrant workers still remain largely outsiders where these measures are concerned. Special attention should be given to this disadvantaged group in terms of social rights and healthcare. Preparedness plans should be improved to deal with contagious pathogens involving the seasonal migrant population. PMID- 15827371 TI - Harmonisation of the acute respiratory infection reporting system in the Czech Republic with the European community networks. AB - Respiratory virus activity is detected in Europe each winter, yet the precise timing and size of this activity is highly unpredictable. The impact of influenza infection and/or acute respiratory infection in European countries is continuously monitored through a variety of surveillance systems. All of these sources of information are used to assess the nature and extent of activity of influenza and other respiratory viruses, and to offer guidance on the prevention and control of morbidity and mortality due to influenza at a local, national and international level. The early warning system for a forthcoming influenza epidemic is mainly based on the use of a set of thresholds. In the Czech Republic, the acute respiratory infection (ARI) reporting system, with automated data processing, uses a statistical model for the early detection of unusual increased rates of the monitored indicators. The collected data consists of the number of ARI, the number of complications due to ARI and the population registered with the reporting general practitioners and paediatricians, all collected separately in five age groups. To improve the reporting system in the Czech Republic, clinical data on the weekly incidence of influenza-like illness (ILI) within the same population and the same age groups was started in January 2004. These data fit the European Commission's recently adopted ILI case definition and allows a better comparison of data with other countries in Europe, in particular those participating in EISS (European Influenza Surveillance Scheme). PMID- 15827372 TI - Suspected SARS patients hospitalised in French isolation units during the early SARS epidemic: the French experience. AB - During the SARS epidemic, many patients were screened according to WHO criteria but never went on to develop SARS. In May 2003, early in the epidemic, we conducted a retrospective study to describe suspected SARS patients hospitalised in France and compared them with documented cases of patients with SARS to evaluate the screening strategy. A total of 117 patients were studied. Only 3.4% had been in close contact with a SARS patient but 73.5% came from an affected area. 67.5% had fever and respiratory symptoms on their admission to hospital. 49.6% had fever and non specific symptoms. Clinical symptoms that were significantly more common among patients with SARS were fever, myalgia, dyspnoea, and nausea or vomiting. Presumed viral fever and respiratory tract infection were the most common diagnosis. Symptoms cannot be distinguished from an early stage of SARS confirming the usefulness of the WHO case definitions in isolation decision to avoid further transmission. PMID- 15827373 TI - Hospital preparedness and management of patients affected by viral haemorrhagic fever or smallpox at the Lazzaro Spallanzani Institute, Italy. AB - The US cases of anthrax in 2001 and the recent severe acute respiratory syndrome outbreak have heightened the need for preparedness and response to naturally emerging and re-emerging infections or deliberately released biological agents. This report describes the response model of the Istituto Nazionale per le Malattie Infettive Lazzaro Spallanzani (INMI), Rome, Italy for managing patients suspected of or affected by smallpox or viral haemorrhagic fever (VHF) either in the context of an intentional release or natural occurrence. The INMI is Italy's leading hospital in its preparedness and response plan to bioterrorism-related infectious agents. All single and double rooms of INMI are equipped with negative air pressure, sealed doors, high efficiency particulate air (HEPA) filters and a fully-equipped anteroom; moreover, a dedicated high isolation unit with a laboratory next door for the initial diagnostic assays is available for admission of sporadic patients requiring high isolation. For patient transportation, two fully equipped ambulances and two stretcher isolators with a negative pressure section are available. Biomolecular and traditional diagnostic assays are currently performed in the biosafety level 3/4 (BSL 3/4) laboratories. Continuing education and training of hospital staff, consistent application of infection control practices, and availability of adequate personnel protective equipment are additional resources implemented for the care of highly infectious patients and to maintain the readiness of an appropriately trained workforce to handle large scale outbreaks. PMID- 15827374 TI - A new simplified catalyzed signal amplification system for minimizing non specific staining in tissues with supersensitive immunohistochemistry. AB - We investigated non-specific staining in a catalyzed reporter deposition (CARD) reaction and improved its blocking methods in supersensitive immunohistochemistry, based on our simplified catalyzed signal amplification (CSA) system (Hasui et al. 2002). In the CARD reaction using biotinyl tyramide, non-specific staining could be reduced by pretreatment with a casein solution or 3% bovine serum albumin (BSA)-phosphate buffer saline (PBS) with 0.1% Tween 20. In the CARD reaction using FITC-labeled tyramide, non-specific staining could be blocked by pretreatment with 0.3% BSA-PBS with 0.1% Tween 20 or 3% polyethylene glycol-PBS with 01% Tween 20. Thus, our new simplified CSA system features: 1) destruction of the endogenous peroxidase activity; 2) blocking of the nonspecific reaction of the primary antibody; 3) a primary antibody reaction; 4) blocking of the non-specific reaction of the polymer reagent by casein treatment; 5) a polymer reaction; 6) blocking of the non-specific reaction of CARD reaction by casein treatment; 7) a CARD reaction; and 8) detection of deposited tyramide. This new system proved useful for detecting an extremely low amount of antigen in the endogenous biotin-rich tissues such as the gastrointestinal tract and liver. By this method, the Ki67 antigen in the G1 phase cell cycle could be detected and a metabolic disorder of the Ki67 antigen was implicated in a carcinoid tumor in the stomach. We believe that this new simplified CSA system represents a new standard of supersensitive immunohistochemistry for use in light-microscopic investigation. PMID- 15827375 TI - Receptors and transporter for serotonin in Merkel cell-nerve endings in the rat sinus hair follicle. An immunohistochemical study. AB - Serotonin (5-HT) has been a candidate for neurotransmitters in cutaneous type I mechanoreceptors (i.e., Merkel cell-nerve endings). Although recent electrophysiological studies have suggested the presence of the 5-HT2 and 3 receptors in the Merkel cell-nerve endings, the histological localization of these receptors are obscure. We thus immunohistochemically examined the presence of 5-HT1, 2, 3 receptors in Merkel cell-nerve endings in sinus hair follicles of the rat whisker pad. We also studied the immunohistochemical localization of the 5-HT transporter to confirm the site of 5-HT secretion. For this purpose, we used antibodies for the 5-HT1A, 5-HT1B, 5-HT2A, 5-HT2C and 5-HT3 receptors, and for the 5-HT transporter, as well as antibodies for cytokeratin 20 (as a marker of Merkel cells) and neurofilament H (a marker of type I sensory nerve terminals). The immuno-stained sections were analyzed under a laser-scanning microscope. It was found that the sensory nerve terminals in the Merkel cell-nerve endings showed strong positive immunoreactions of 5-HT1A and 1B receptors but not 5-HT2A, 2C, and 3 receptors. Furthermore, both the Merkel cells and related axon terminals showed strong immunoreactions of the 5-HT transporter. These findings support the idea that 5-HT molecules are released from the Merkel cells during mechanical reception and indirectly regulate neural actions of sensory neurons via 5-HT1 receptors. The localization of the 5-HT transporter found in this study also suggests a possibility that axon terminals in the Merkel cell-nerve endings also release 5-HT. PMID- 15827376 TI - The occurrence of nitric oxide synthase-containing axonal baskets surrounding large neurons in rat dorsal root ganglia after sciatic nerve ligation. AB - To clarify the possible role of nitric oxide (NO) induced in primary sensory neurons after peripheral axotomy, NO synthase (NOS) immunohistochemistry was carried out on rat L5 dorsal root ganglia after sciatic nerve ligation. The results were compared with the expression of 27-kDa heat shock protein (HSP27), a neuroprotective molecule. In intact animals, NOS-immunoreactive neurons represented about 2% of all dorsal root ganglion (DRG) neurons, whereas HSP27 immunoreactive neurons comprised about 14%. After sciatic nerve ligation, both neurons increased, in number and immunoreactivity, reaching a maximum at 2 weeks, when NOS- and HSP27-immunoreactive neurons represented about 33 and 66%, respectively. NOS-immunoreactive neurons then remained unchanged until 7 weeks although HSP27-immunoreactive neurons showed a slight decline. The increased NOS immunoreactive neurons were preferentially small (100-500 microm(2)) and coexpressed with HSP27 (about 87%). On the other hand, in the proximal stump of sciatic nerves, numerous NOS-immunoreactive fibers with a regenerative profile appeared transiently (2-4 weeks). At higher magnification, an axonal sprout from the NOS-immunoreactive small DRG neurons was found to form a basket-like structure (or basket) mostly around the cell body of NOS-negative large neurons. Retrograde labeling with a fluorescent tracer showed that both neurons sent peripheral axon collaterals to the sciatic nerve. The appearance of this unique structure was most prominent after depletion of the NOS-immunoreactive regenerating fibers in the sciatic nerve (at 7-9 weeks). The findings suggest that NO might be involved in not only axonal regeneration but also the rewiring of two classes of DRG neurons after peripheral nerve injury. PMID- 15827377 TI - Effects of a phenolic compound, resveratrol, on the renal function and costimulatory adhesion molecule CD86 expression in rat kidneys with ischemia/reperfusion injury. AB - Recent studies have suggested that an ischemia/reperfusion (I/R) injury enhances the expression of costimulatory adhesion molecules on the vascular endothelium. In the present study, we investigated the protective effects of resveratrol, a phenolic product, on the renal function and expression of CD86 in rat kidneys with I/R injury. Wistar rats were divided into four groups; 1) an I/R group with right nephrectomy and 1-hour clamping of the left renal pedicle; 2) a vehicle group, I/R plus 10% ethanol (0.1 ml/kg/day) administered by intra-peritoneal injection from day -1 through to 7; 3) a resveratrol group, I/R plus 4 mg/kg/day of resveratrol; and 4) a sham group. Blood samples were obtained via the tail vein at 1 day before, and 1, 3, and 7 days after the operation (day 0) for the measurement of serum creatinine (Scr) levels. The expression of CD86 protein was analyzed by immunofluorescence staining, and the level of CD86 messenger RNA (mRNA) was evaluated quantitatively by a real-time reverse transcription polymerase chain reaction (RT-PCR) in the renal cortex at day 3. Scr levels of the resveratrol group were significantly lower than those of the I/R and vehicle groups on days 1 and 3 after the operation. From the immunohistochemical study, the expression of CD86 in the glomerular endothelium and peritubular vessels was found to be attenuated in the resveratrol group compared with the I/R or vehicle group. In the resveratrol group, the CD86 mRNA level was significantly lower than that in the I/R or vehicle group, and it was significantly decreased by about one fifth of that in the sham group. Our results suggest that resveratrol markedly reduces renal dysfunction and attenuates the mRNA and protein expression of CD86 following I/R injury. PMID- 15827378 TI - Three-dimensional ultrastructure of the brush border glycocalyx in the mouse small intestine: a high resolution scanning electron microscopic study. AB - The three-dimensional ultrastructure of the filamentous glycocalyx of the brush border in the mouse small intestine was successfully demonstrated by high resolution scanning electron microscopy (SEM). The specimens were fixed with 2% glutaraldehyde in a 0.1M phosphate buffer (pH 7.4), and rinsed with buffered solutions with differently adjusted pH values (pH 3.0, 7.0 or 11.0). They were then osmicated, dried, spatter-coated with gold (1.0-1.5 nm), and observed under a high resolution SEM. The glycocalyx on the luminal surface of the intestinal villi covered the top of the microvilli of the epithelial cells and were well preserved in the specimens treated with an alkaline buffer (pH 11.0). The glycocalyx was observed as filamentous structures, 7 to 15 nm thick in diameter. These filaments repeatedly branched and anastomosed with neighboring ones to form an actual network or plexus as a whole, in contrast with superimposed images in transmission electron microscopy (TEM) which suggested that such anastomoses were pseudo-networks. The filaments thickened globularly at the sites of the filament bifurcation or branching. On the other hand, specimens rinsed with an acid or neutral buffer showed no glycocalyx on their microvilli, whose naked top had knob like structures. Thus, the pH values of the washing buffer solutions were considered to affect the preservation of the surface coat due to molecular characteristics. PMID- 15827379 TI - An immunocytochemical study of calbindin-D28K in laminae I and II of the dorsal horn and spinal ganglia in the chicken with special reference to the relation to substance P-containing primary afferent neurons. AB - The localization of calbindin-D28K (CB) was studied immunocytochemically in laminae I and II of the dorsal horn and in spinal ganglia in the chicken, and compared with the distribution of substance P (SP) using double immunolabeling. At the light microscopic level, CB immunoreactivity was observed most intensely in the lamina II using the avidin-biotinylated peroxidase complex (ABC) and immunofluorescence methods. At the electron microscopic level using the ABC method, CB immunoreactivity was observed in the following three neuronal elements: 1) the scalloped central terminal with many dense-cored vesicles (DCVs) in the synaptic glomerulus; 2) some vesicle-containing dendrites (VCDs) inside or outside the synaptic glomerulus; and 3) some axon terminals outside the synaptic glomerulus. The CB-immunoreactive (IR) VCDs in the synaptic glomerulus often formed reciprocal synapses with the central terminal. Strong immunoreactivity was observed at the postsynaptic membrane of CB-IR elements. Double immunofluorescence and immunolabeling methods at the electron microscopic level showed that CB and SP colocalized in the scalloped central terminal with DCVs of the synaptic glomerulus. Almost all SP-IR neurons in the spinal ganglion revealed the coexistence of CB in serial sections in the chicken. In light of previous biochemical and physiological reports, our findings suggest that CB - coexisting with SP - plays an important role in the control of pain transmission through its strong Ca(2+)-buffering action in the chicken. PMID- 15827381 TI - The structure of C-banded human metaphase chromosomes as observed by atomic force microscopy. AB - The ultrastructure of C-banded human metaphase chromosomes was studied by the combined use of light microscopy and atomic force microscopy (AFM). Light microscopy of the C-banded chromosomes showed that the centromeric regions of all chromosomes except the Y chromosome were positively stained. AFM further revealed that the C-positive region was higher than the C-negative region. The area of the C-positive region was specific depending on each chromosome; it ranged from the centromere to the proximal end of the long arm in chromosome 1, while it was restricted to the centromere in chromosomes 2 and 3. At higher magnification, chromatin fibers about 50 nm thick were clearly shown in the entire length of the chromosomes. In the C-positive region, these chromatin fibers were densely packed, while chromatin fibers were loosely packed with gentle twisting in the C negative region. These AFM findings suggest that certain factors related to the chromatin fiber compaction remain in the C-positive region even after successive C-banging treatment. PMID- 15827380 TI - Participation of autophagy in the degeneration process of rat hepatocytes after transplantation following prolonged cold preservation. AB - Cold ischemia-warm reperfusion injury of liver grafts has been investigated thoroughly, but its underlying mechanism remains poorly understood. Here we show that autophagy is involved not only during cold preservation but also during warm reperfusion following transplantation. Immunohistochemistry using an antibody against LC3, a microtubule associated protein 1 light chain 3 and a marker of autophagosomes, showed dot-like weak staining in hepatocytes of rat liver grafts during cold preservation. Since University of Wisconsin solution for graft preservation lacks amino acids, the induction of autophagy in hepatocytes was similar to that under starvation conditions. Intense immunopositive punctate structures were detected abundantly in the hepatocytes 30 min after the beginning of reperfusion. LC3-positive granules were often co-localized in ED2-positive Kupffer cells at 60 min of the reperfusion phase. The molecular form of LC3 was mainly LC3-II, a membrane-bound form, during reperfusion, especially at 30 min of the phase. Electron microscopic examination demonstrated numerous vacuolar structures in hepatocytes at 30 min of the reperfusion period, while some hepatocytes with such vacuolar structures were present in the sinusoidal lumen. At the late stage of the reperfusion period, Kupffer cells contained phagocytosed cells that possessed numerous autophagic vacuoles/autolysosomes and nuclei with condensed chromatin. Our results showing the presence of autophagic vacuoles/autolysosomes in hepatocytes of liver grafts after the start of reperfusion suggest that warm reperfusion acted as a stress stimulus to hepatocytes. Moreover, the stress response of hepatocytes may be involved in their degeneration process. PMID- 15827382 TI - Influence of estrus synchronization of prepubertal gilts on embryo quality. AB - Synchronization and superovulation are commonly used to obtain large numbers of embryos for experimental and practical purposes. This study compared the number, quality, and in vitro development of embryos recovered from gilts following single or double estrus synchronization and superovulation. Prepubertal gilts from the single synchronization group were injected with 1500 I.U. PMSG and 1000 I.U. hCG 72 h later. The double synchronized group of gilts was treated with 750 I.U. PMSG and 500 I.U. hCG 72 h later. After 17 days, 1500 I.U. PMSG followed by 1000 I.U. hCG was administered. Five days after insemination embryos were recovered and cultured for 6 days. Both single and double hormonal stimulation schedules resulted in recovery of elevated numbers of embryos (28.4 and 23.4 vs. 11.3; p<0.01 and pT mutation in congenital fibrosis of extraocular muscles type 1 and 3. AB - PURPOSE: To determine, in Taiwanese patients, the genetic basis of congenital fibrosis of the extraocular muscles (CFEOM) type 1 and 3, a group of diseases characterized by congenital restrictive ophthalmoplegia affecting muscles innervated by the oculomotor nerve. METHODS: Linkage analysis with microsatellite markers at chromosome 12q and direct sequence analysis of the KIF21A gene were performed on three families and one sporadic CFEOM case. RESULTS: Two of the families were classified by clinical criteria as CFEOM1 and another family as CFEOM3. All three families were shown to be linked to the 12q CFEOM1 locus. Sequence analysis disclosed a heterozygous c.2860C>T mutation in the KIF21A gene in all families and in the sporadic case. Affected family members were further confirmed by a BsrDI polymorphism. CONCLUSIONS: CFEOM is present in Chinese populations. Both CFEOM1 and CFEOM3 can be caused by the same mutation at the KIF21A gene. The occurrence of this mutation in different ethnic groups suggests a mutation hot spot. PMID- 15827545 TI - Immunolocalization of ciliary neurotrophic factor receptor alpha (CNTFRalpha) in mammalian photoreceptor cells. AB - PURPOSE: To characterize the site of expression of the alpha subunit of the receptor for ciliary neurotrophic factor (CNTFRalpha) in the retina of a variety of mammalian species, and determine whether CNTFRalpha is localized to photoreceptor cells. METHODS: The cellular distribution of CNTFRalpha(protein) was examined by immunocytochemistry in the adult retinas of several mammalian species that included mouse, rat, dog, cat, sheep, pig, horse, monkey, and human. Developing retinas from 3-day-old and 6-day-old rats were also included in this study. The molecular weight of CNTFRalpha in rat, dog, cat, pig, and human retinas was determined by immunoblotting. RESULTS: CNTFRalpha immunolabeling was present in the retina of all species. A common pattern was observed in all species, and represented labeling of the nerve fiber layer (NFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), and outer plexiform layer (OPL). CNTFRalpha did not immunolocalize to photoreceptor cells in both adult and developing rodent retinas, but was consistently observed in both rods and cones of non-rodent species. The molecular weight of CNTFRalpha in mammalian retinas was approximately 61-64 kDa. CONCLUSIONS: These findings highlight a significant difference in the expression of CNTFRalpha in the retina of rodent and non-rodent mammalian species. The expression of CNTFRalpha by rods and cones in non-rodent species may suggest a direct mechanism of action if CNTF administration results in photoreceptor rescue. PMID- 15827547 TI - Arginine hydrochloride enhances the dynamics of subunit assembly and the chaperone-like activity of alpha-crystallin. AB - PURPOSE: Alpha-crystallin, a major eye lens protein, bears homology with small heat shock proteins (sHsps) and exhibits molecular chaperone-like activity. Structural perturbation by temperature or low concentrations of denaturants leads to enhancement of its chaperone-like activity. We have earlier demonstrated similar enhancement of chaperone-like activity using biologically compatible solutes such as arginine hydrochloride and aminoguanidine. The purpose of the present study is to get an insight into the mechanism of the arginine induced enhancement of chaperone-like activity of alpha-crystallin. METHODS: The effect of arginine hydrochloride on the chaperone-like activity of alpha-crystallin at 25 degrees C was studied using DTT induced aggregation of insulin as a model system. Changes in the accessibility of the thiol group near the end of the alpha crystallin domain in the absence and the presence of arginine hydrochloride were studied using dithiobisnitrobenzoic acid. Fluorescence resonance energy transfer studies were performed to investigate changes in the dynamics of the subunit assembly. Urea induced denaturation studies of alpha-crystallin were carried out to investigate structural destabilization of alpha-crystallin, if any, in the presence of arginine hydrochloride. RESULTS: Arginine hydrochloride increases the chaperone-like activity of alpha-crystallin several fold towards DTT induced aggregation of insulin at room temperature. Our study shows that both the extent and the rate of accessibility of the thiol group are increased in the presence of arginine. Fluorescence resonance energy transfer experiments show that arginine hydrochloride significantly increases the subunit exchange between the oligomers of alpha-crystallin. Arginine induced structural perturbation and loosening of subunit assembly of alpha-crystallin leads to overall destabilization of the protein as reflected by the urea denaturation study. CONCLUSIONS: Arginine perturbs the tertiary and quaternary structure of alpha-crystallin and enhances the dynamics of the subunit assembly leading to enhanced chaperone-like activity. Thus, in addition to size, surface hydrophobicity, and charge distribution, the dynamics of the subunit assembly appears to be one of the critical factors that can modulate the chaperone activity. PMID- 15827548 TI - Multilayered retinal microglial response to optic nerve transection in rats. AB - PURPOSE: Microglia normally exist in several layers across the retinal thickness. When retinal ganglion cells undergo apoptosis after lesion to their axons, microglial cells proliferate and promptly clear the debris. We have previously reported on the phagocytic response following optic nerve axotomy. Here, we present how microglial cells of deeper retinal layers are affected by transection of the optic nerve. METHODS: Normal and reactive microglia in the retina of adult rats whose optic nerves had been lesioned were labeled by using antibodies OX42 and ED1. Analysis of the time course (between 1 and 180 days post-axotomy) of appearance and distribution of microglial cells in the retinal layers was performed. RESULTS: In normal retinas, microglia were found in the ganglion cell layer (GCL), the superficial inner nuclear layer (INL) and the outer plexiform layer (OPL). Increases in numbers of microglia occurred maximally in the GCL at day 12 post-axotomy. Increases were also detected in the superficial INL. The proliferation of these phagocytic cells led to their highest numbers in the more central eccentricities in the two most superficial layers. Microglia in the OPL remained undisturbed. Microglial normal histology is restored over a period of 6 months as dying ganglion cells disappear. CONCLUSIONS: Histological characteristics of normal rat retinal microglia are uniform across different retinal eccentricities for each of the three laminae where they exist. Retinal microglia of various layers respond to optic nerve damage. Their increase in numbers and morphological transformation follow retinal ganglion cell death. Their morphology, density, and layered distribution slowly return to normal, confirming that retinal ganglion cells, or their densities, do not play any role in dictating microglial distribution within the different retinal layers. PMID- 15827549 TI - Pharmacokinetics and metabolism of ifosfamide in relation to DNA damage assessed by the COMET assay in children with cancer. AB - The degree of damage to DNA following ifosfamide (IFO) treatment may be linked to the therapeutic efficacy. The pharmacokinetics and metabolism of IFO were studied in 19 paediatric patients, mostly with rhabdomyosarcoma or Ewings sarcoma. Ifosfamide was dosed either as a continuous infusion or as fractionated doses over 2 or 3 days. Samples of peripheral blood lymphocytes were obtained during and up to 96 h after treatment, and again prior to the next cycle of chemotherapy. DNA damage was measured using the alkaline COMET assay, and quantified as the percentage of highly damaged cells per sample. Samples were also taken for the determination of IFO and metabolites. Pharmacokinetics and metabolism of IFO were comparable with previous studies. Elevations in DNA damage could be determined in all patients after IFO administration. The degree of damage increased to a peak at 72 h, but had returned to pretreatment values prior to the next dose of chemotherapy. There was a good correlation between area under the curve of IFO and the cumulative percentage of cells with DNA damage (r2=0.554, P=0.004), but only in those patients receiving fractionated dosing. The latter patients had more DNA damage (mean+/-s.d., 2736+/-597) than those patients in whom IFO was administered by continuous infusion (1453+/-730). The COMET assay can be used to quantify DNA damage following IFO therapy. Fractionated dosing causes a greater degree of DNA damage, which may suggest a greater degree of efficacy, with a good correlation between pharmacokinetic and pharmacodynamic data. PMID- 15827550 TI - Quantitative relationship between functionally active telomerase and major telomerase components (hTERT and hTR) in acute leukaemia cells. AB - Functionally active telomerase is affected at various steps including transcriptional and post-transcriptional levels of major telomerase components (hTR and human telomerase reverse transcriptase (hTERT)). We therefore developed a rapid and sensitive method to quantify hTERT and its splicing variants as well as the hTR by a Taqman real-time reverse transcriptase-polymerase chain reaction to determine whether their altered expression may contribute to telomere attrition in vivo or not. Fresh leukaemia cells obtained from 38 consecutive patients were used in this study. The enzymatic level of telomerase activity measured by TRAP assay was generally associated with the copy numbers of full length hTERT+alpha+beta mRNA (P=0.0024), but did not correlate with hTR expression (P=0.6753). In spite of high copy numbers of full-length hTERT mRNA, telomerase activity was low in some cases correlating with low copy numbers of hTR, raising the possibility that alteration of the hTR : hTERT ratio may affect functionally active telomerase activity in vivo. The spliced nonactive hTERT mRNA tends to be lower in patients with high telomerase activity, suggesting that this epiphenomenon may play some role in telomerase regulation. An understanding of the complexities of telomerase gene regulation in biologically heterogeneous leukaemia cells may offer new therapeutic approaches to the treatment of acute leukaemia. PMID- 15827551 TI - Effects of cisplatin on olfactory function in cancer patients. AB - A prospective analysis of olfaction was performed in 21 patients receiving cisplatin. A reduction in olfactory function was noted in only one patient. Hearing impairment was documented in nine patients, none of whom had impaired sense of smell. We conclude that cisplatin has no major deleterious effect on olfactory function at doses which cause hearing impairment. PMID- 15827552 TI - Whole grain consumption and risk of colorectal cancer: a population-based cohort of 60,000 women. AB - We examined prospectively the association between whole grain consumption and colorectal cancer risk in the population-based Swedish Mammography Cohort. A total of 61,433 women completed a food-frequency questionnaire at baseline (1987 1990) and, through linkage with the Swedish Cancer Registry, 805 incident cases of colorectal cancer were identified during a mean follow-up of 14.8 years. High consumption of whole grains was associated with a lower risk of colon cancer, but not of rectal cancer. The multivariate rate ratio (RR) of colon cancer for the top category of whole grain consumption (> or = 4.5 servings day(-1)) compared with the bottom category (<1.5 servings day(-1)) was 0.67 (95% confidence interval (CI), 0.47-0.96; P-value for trend=0.06). The corresponding RR after excluding cases occurring within the first 2 years of follow-up was 0.65 (95% CI, 0.45-0.94; P-value for trend=0.04). Our findings suggest that high consumption of whole grains may decrease the risk of colon cancer in women. PMID- 15827553 TI - Long-term protective effect of high-risk human papillomavirus testing in population-based cervical screening. AB - We prospectively evaluated the 5-year predictive values of adding high-risk human papillomavirus (hrHPV) testing to cytology for the detection of > or = cervical intraepithelial neoplasia (CIN)3 lesions in a population-based cohort of 2810 women. At baseline, nine (0.3%) women had prevalent lesions > or = CIN3, all being hrHPV positive. After 5 years of follow-up, four (6.5%) of the 62 hrHPV positive women with normal cytology developed lesions > or = CIN3, vs only one (0.05%) of the 2175 hrHPV-negative women with normal cytology. High-risk human papillomavirus testing or combined screening revealed a much higher sensitivity, at the cost of a small decrease in specificity, and a higher negative predictive value for the detection of lesions > or = CIN3 till the next screening round (5 years) than cytology alone. PMID- 15827554 TI - Cancer survival in Kampala, Uganda. AB - Epidemiological data on the occurrence of cancer in sub-Saharan Africa are sparse, and population-based cancer survival data are even more difficult to obtain due to various logistic difficulties. The population-based Cancer Registry of Kampala, Uganda, has followed up the vital status of all registered cancer patients with one of the 14 most common forms of cancer, who were diagnosed and registered between 1993 and 1997 in the study area. We report 5-year absolute and relative survival estimates of the Ugandan patients and compare them with those of black American patients diagnosed in the same years and included in the SEER Program of the United States. In general, the prognosis of cancer patients in Uganda was very poor. Differences in survival between the two patient populations were particularly dramatic for those cancer types for which early diagnosis and effective treatment is possible. For example, 5-year relative survival was as low as 8.3% for colorectal cancer and 17.7% for cervical cancer in Uganda, compared with 54.2 and 63.9%, respectively, for black American patients. The collection of good-quality follow-up data was possible in the African environment. The very poor prognosis of Ugandan patients is most likely explained by the lack of access to early diagnosis and treatment options in the country. On the policy level, the results underscore the importance of the consistent application of the national cancer control programme guidelines as outlined by the World Health Organization. PMID- 15827555 TI - Serum cholesterol and testicular cancer incidence in 45,000 men followed for 25 years. AB - In a 25-year follow-up study of 44,864 men with measured serum cholesterol levels, the testicular cancer hazard ratios for the serum cholesterol categories 5.7-6.9 and > or = 7.0 mmol l(-1) vs the reference category (<5.7 mmol l(-1)) were 1.3 and 4.5, respectively; P-value for trend=0.005. This highly significant association suggests that high-serum cholesterol is a risk factor for testicular cancer. PMID- 15827556 TI - Human papillomavirus infection in women who develop high-grade cervical intraepithelial neoplasia or cervical cancer: a case-control study in the UK. AB - Human papillomavirus (HPV) testing might identify older women who could be withdrawn from the cervical screening programme, or require less frequent screening. A case-control study using the United Kingdom cervical screening population was set up to help address this issue. Cases comprised 575 women who developed cervical intraepithelial neoplasia (CIN) grade 2 or worse over a 13 year period following a cytologically normal baseline smear, and were stratified by age group ('under 20', '20-39' and 40 years or over). Controls (n=601) were women who remained disease free over this interval and were the same age on average as cases. DNA was extracted from the baseline smears and tested for HPV by PCR using GP5+/6+ consensus primers. HPV+ samples were tested for HPV types 16 and 18 using specific PCR primers. In all, 27.0% of cases tested positive for HPV at baseline, compared with 15.4% of controls (odds ratio (OR)=2.00; 95% confidence interval (CI), 1.50-2.68). Among women aged 40 years or over, the OR for HPV 16 was 8.95 (95% CI, 2.63-30.4). These results support the need for further cervical screening of HPV- older women, as many of the cases were HPV- at baseline. PMID- 15827557 TI - Garlic arrests MDA-MB-435 cancer cells in mitosis, phosphorylates the proapoptotic BH3-only protein BimEL and induces apoptosis. AB - Components of garlic (Allium sativum) can cause disruption of microtubules, cell cycle arrest, and apoptosis in cancer cells. We show here that a water-soluble extract of garlic arrested MDA-MB-435 cancer cells in mitosis and caused apoptosis. The proapoptotic BH3-only, bcl-2 family protein BimEL, which in healthy cells can be tightly sequestered to the microtubule-associated dynein motor complex, was modified after garlic treatment. The main effect of garlic on BimEL was a considerable increase in a phosphorylated form of the protein. This phosphorylation(s), probably partly dependent on c-jun N-terminal kinase activity, promoted mitochondrial localisation of BimEL. Furthermore, inhibition of extracellular signal-regulated kinases 1/2 increased the amount of another form of BimEL present in the mitochondrial cellular fraction. Treatment of cells with the garlic compound diallyl disulphide had similar effects on BimEL. The results indicate that the apoptotic effect of garlic and a combination of garlic and the inhibitor of extracellular signal-regulated kinases 1/2 in MDA-MB-435 cells partly is due to modifications that are necessary for translocation of the proapoptotic protein BimEL to mitochondria where it executes its proapoptotic function. PMID- 15827558 TI - Clinical significance of heparin-binding epidermal growth factor-like growth factor in peritoneal fluid of ovarian cancer. AB - Epidermal growth factor receptor (EGFR) has been implicated in tumour growth and extension of ovarian cancer. Peritoneal fluid in ovarian cancer patients contains various growth factors that can promote tumour growth and extension. In order to investigate the clinical significance of EGFR ligands as activating factors of ovarian cancer, we examined the cell proliferation-promoting activity and the level of EGFR ligands in peritoneal fluid obtained from 99 patients. Proliferation-promoting activity in peritoneal fluid from 63 ovarian cancer patients (OVCA) was much higher than peritoneal fluid from 18 ovarian cyst patients (OVC) and 18 normal ovary patients (NO), and the activity was suppressed only by antibodies against EGFR or heparin-binding epidermal growth factor (HB EGF). A large difference was observed in the level of EGFR ligands between HB-EGF and TGF-alpha or amphiregulin. The concentration of HB-EGF in OVCA significantly increased compared to that in OVC or NO (P<0.01). No significant difference in the concentration of TGF-alpha and amphiregulin was found between the OVCA and NO or OVC groups. In peritoneal fluid, HB-EGF is sufficiently elevated to activate cancer cells even at an early stage of OVCA. These results suggested that HB-EGF in peritoneal fluid might play a key role in cell survival and in the proliferation of OVCA. PMID- 15827560 TI - The effects of conjugated linoleic acid supplementation on immune function in healthy volunteers. AB - OBJECTIVE: To assess the effects of dietary supplementation using two isomeric blends of conjugated linoleic acid (CLA) on immune function in healthy human volunteers. DESIGN: Double-blind, randomised, placebo-controlled intervention trial. SUBJECTS AND INTERVENTION: A total of 55 healthy volunteers (n=20 males, n=35 females) were randomised into one of three study groups who received 3 g/day of a fatty acid blend containing a 50:50 cis-9, trans-11: trans-10, cis-12 CLA isomer blend (2 g CLA), and 80:20 cis-9, trans-11: trans-10, cis-12 (80:20) CLA isomer blend (1.76 g CLA) or linoleic acid (control, 2 g linoleic acid) for 8 weeks. RESULTS: Supplementation with the 80:20 CLA isomer blend significantly (P< or =0.05) enhanced PHA-induced lymphocyte proliferation. CLA decreased basal interleukin (IL)-2 secretion (P< or =0.01) and increased PHA-induced IL-2 and tumor necrosis factor alpha (TNF(alpha)) production (P< or =0.01). However, these effects were not solely attributable to CLA as similar results were observed with linoleic acid. CLA supplementation had no significant effect on peripheral blood mononuclear cells IL-4 production, or on serum-soluble intercellular adhesion molecule-1 (sICAM-1) or plasma prostaglandin E2 (PGE2) or leukotreine B4 (LTB4) concentrations. CONCLUSIONS: This study shows that CLA supplementation had a minimal effect on the markers of human immune function. Furthermore, supplementation with CLA had no immunological benefit compared with linoleic acid. PMID- 15827559 TI - Fluorescence in situ hybridisation analysis of chromosomal aberrations in gastric tissue: the potential involvement of Helicobacter pylori. AB - In this series of experiments, a novel protocol was developed whereby gastric cells were collected using endoscopic cytology brush techniques, and prepared, such that interphase fluorescence in situ hybridization (FISH) could be performed. In total, 80 distinct histological samples from 37 patients were studied using four chromosome probes (over 32,000 cells analysed). Studies have previously identified abnormalities of these four chromosomes in upper GI tumours. Using premalignant tissues, we aimed to determine how early in Correa's pathway to gastric cancer these chromosome abnormalities occurred. Aneuploidy of chromosomes 4, 8, 20 and 17(p53) was detected in histologically normal gastric mucosa, as well as in gastritis, intestinal metaplasia, dysplasia and cancer samples. The levels of aneuploidy increased as disease severity increased. Amplification of chromosome 4 and chromosome 20, and deletion of chromosome 17(p53) were the more common findings. Hence, a role for these abnormalities may exist in the initiation of, and the progression to, gastric cancer. Helicobacter pylori infection was determined in premalignant tissue using histological analysis and PCR technology. Detection rates were comparable. PCR was used to subtype H. pylori for CagA status. The amplification of chromosome 4 in gastric tissue was significantly more prevalent in H. pylori-positive patients (n=7) compared to H. pylori-negative patients (n=11), possibly reflecting a role for chromosome 4 amplification in H. pylori-induced gastric cancer. The more virulent CagA strain of H. pylori was associated with increased disease pathology and chromosomal abnormalities, although numbers were small (CagA+ n=3, CagA- n=4). Finally, in vitro work demonstrated that the aneuploidy induced in a human cell line after exposure to the reactive oxygen species (ROS) hydrogen peroxide was similar to that already shown in the gastric cancer pathway, and may further strengthen the hypothesis that H. pylori causes gastric cancer progression via an ROS-mediated mechanism. PMID- 15827561 TI - Mitochondrial DNA haplogroup K is associated with a lower risk of Parkinson's disease in Italians. AB - It has been proposed that European mitochondrial DNA (mtDNA) haplogroups J and K, and their shared 10398G single-nucleotide polymorphism (SNP) in the ND3 gene, are protective from Parkinson's disease (PD). We evaluated the distribution of the different mtDNA haplogroups in a large cohort of 620 Italian patients with adult onset (>50, <65 years of age) idiopathic PD vs two groups of ethnic-matched controls. Neither the frequencies of haplogroup J nor that of 10398G were significantly different. However, the frequency of haplogroup K was significantly lower in PD. Stratification by sex and age indicated that the difference in the distribution of haplogroup K was more prominent in >50 year old males. In spite of the common 10398G SNP, haplogroups J and K belong to widely diverging mitochondrial clades, a consideration that may explain the different results obtained for the two haplogroups in our cohorts. Our study suggests that haplogroup K might confer a lower risk for PD in Italians, corroborating the idea that the mitochondrial oxidative phosphorylation pathway is involved in the susceptibility to idiopathic PD. PMID- 15827562 TI - Protein studies in dysferlinopathy patients using llama-derived antibody fragments selected by phage display. AB - Mutations in dysferlin, a member of the fer1-like protein family that plays a role in membrane integrity and repair, can give rise to a spectrum of neuromuscular disorders with phenotypic variability including limb-girdle muscular dystrophy 2B, Myoshi myopathy and distal anterior compartment myopathy. To improve the tools available for understanding the pathogenesis of the dysferlinopathies, we have established a large source of highly specific antibody reagents against dysferlin by selection of heavy-chain antibody fragments originating from a nonimmune llama-derived phage-display library. By utilizing different truncated forms of recombinant dysferlin for selection and diverse selection methodologies, antibody fragments with specificity for two different dysferlin domains could be identified. The selected llama antibody fragments are functional in Western blotting, immunofluorescence microscopy and immunoprecipitation applications. Using these antibody fragments, we found that calpain 3, which shows a secondary reduction in the dysferlinopathies, interacts with dysferlin. PMID- 15827563 TI - Haplotype construction of the FRDA gene and evaluation of its role in type II diabetes. AB - A GAA-repeat in the X25 gene is causing Friedreich's ataxia (FRDA), a common neurodegenerative disease and >20% of FRDA patients develop type II diabetes (T2D). Linkage has previously been detected between T2D and chromosome 9p13-q21, the region that harbours the X25 gene, but association studies of this gene in T2D have been contradicting. Here, we examined whether genetic variation in the X25 gene is associated with risk for T2D. The GAA-repeat and 18 single nucleotide polymorphisms (SNPs) covering the X25 gene were genotyped in 220 trios in which the affected offspring had abnormal glucose tolerance. Any nominally significant findings were examined in an independent sample consisting of 523 individuals with T2D and 326 healthy controls. Previously reported results were analysed together with our data using a meta-analysis approach. There was no association between the GAA-repeat and T2D susceptibility in our study, which was supported by the meta-analysis including all previous publications. One SNP (rs2498429), 8.2 kb downstream of X25, was nominally associated with T2D in the trios (P=0.02) and showed a trend of association in the same direction in the case-controls (P=0.08; combined permuted P=0.01). Further analysis showed that the nine-marker haplotype containing the rare allele of rs2498429 was nominally associated with T2D in the trios (P<0.01) as well as in the case-controls (P=0.03). In conclusions, this study excludes a role of genetic variation within the X25 gene, but instead suggests that genetic variation downstream the X25 gene, may increase risk for T2D. PMID- 15827564 TI - Univariate and bivariate variance component linkage analysis of a whole-genome scan for loci contributing to bone mineral density. AB - Osteoporosis is a common condition characterized by reduced skeletal strength and increased susceptibility to fracture. The single major risk factor for osteoporosis is low bone mineral density (BMD) and strong evidence exists that genetic factors are in part responsible for an individual's BMD. A cohort of 40 multiplex Caucasian families selected through a proband with osteoporosis was genotyped for microsatellite markers spaced at an average of 10 cM, and linkage to femoral neck (FN), lumbar spine (LS) and trochanter (TR) BMD was analyzed using univariate and bivariate variance component linkage analysis. Maximum univariate multipoint lod-scores were 2.87 on chromosome 1p36 for FN BMD, 1.89 on 6q27 for TR BMD, and 2.15 on 7p15 for LS BMD. Results of bivariate linkage analysis were highly correlated with those of the univariate analysis, although generally less significant, suggesting the possibility that some of these susceptibility loci may exert pleiotropic effects on multiple skeletal sites. PMID- 15827565 TI - Hardy-Weinberg equilibrium in genetic association studies: an empirical evaluation of reporting, deviations, and power. AB - We evaluated the testing and reporting of Hardy-Weinberg equilibrium (HWE) in recent genetic association studies, detected how frequently HWE was violated and estimated the power for HWE testing in this literature. Genetic association studies published in 2002 in Nature Genetics, American Journal of Human Genetics, and American Journal of Medical Genetics were assessed. Data were analyzed on 239 biallelic associations using 154 distinct genotype distribution data sets where HWE could be tested. Any information on HWE was given only for 150 (62.8%) associations (92 (59.7%) data sets). Reanalysis of the data showed significant deviation from HWE in the disease-free controls of 20 associations (13 data sets), but only four of them (two data sets) were admitted in the published articles. Another four deviations (in two data sets) were observed in the combined sample of cases and controls of studies where both cases and controls were diseased, and none were reported in the papers. In all six tested multiallelic associations (six data sets), there was violation of HWE, but this was not admitted in the published articles. Power calculations showed that most studies conforming to HWE simply were largely underpowered to detect HWE deviation; for example, power to detect an inbreeding of magnitude F=0.10 exceeded 80% in only 11 (7%) of the data sets being tested. This empirical evidence suggests that, even in high profile genetics journals, testing and reporting for HWE is often neglected and deviations are rarely admitted in the published reports. Moreover, power is limited for HWE testing in most current genetic association studies. PMID- 15827566 TI - Decrease in the AP-2 DNA-binding activity and in the protein expression of AP-2 alpha and AP-2 beta in frontal cortex of rats treated with lithium for 6 weeks. AB - Lithium chloride (LiCl), when fed to rats for 6 weeks, has been reported to decrease brain mRNA, protein, and activity levels of arachidonic acid (AA) selective cytosolic phospholipase A2 (cPLA2), without affecting secretory sPLA2 or Ca2+-independent iPLA2. We investigated whether transcription factors known to regulate cPLA2 gene expression are modulated by chronic lithium treatment. Male Fischer-344 rats were fed a LiCl-containing diet for 6 weeks to produce a therapeutically relevant brain lithium concentration. Control animals were fed a LiCl-free diet. Using a gelshift assay, we found that LiCl significantly decreased activating protein 2 (AP-2)-binding activity, and protein levels of the AP-2 alpha and AP-2 beta but not of the AP-2 gamma subunits in the frontal cortex. Activating protein 1 (AP-1)-binding activity was increased, whereas glucocorticoid response element, polyoma enhancer activator 3, and nuclear factor kappa B DNA-binding activities were not changed significantly. Since both cPLA2 and AP-2 can be activated by protein kinase C (PKC), we examined the frontal cortex protein levels of PKC alpha and PKC epsilon, as well as AA-dependent PKC activity. The protein levels of PKC alpha and PKC epsilon were decreased significantly, as was AA-dependent PKC activity, in the lithium-treated compared to control rats. Our results suggest that the reported decrease in brain gene expression of cPLA2 by chronic lithium may be mediated by reduced AP-2 transcriptional activity, and that decreased expression of PKC alpha and PKC epsilon contributes to lowering the AP-2 activity. PMID- 15827567 TI - Glycogen synthase kinase-3: a putative molecular target for lithium mimetic drugs. AB - Despite many decades of clinical use, the therapeutic target of lithium remains uncertain. It is recognized that therapeutic concentrations of lithium, through competition with the similarly sized magnesium cation, inhibit the activity of select enzymes. Among these is glycogen synthase kinase-3 (GSK-3). Recent preclinical evidence, including biochemical, pharmacological, genetic, and rodent behavioral models, supports the hypothesis that inhibition of GSK-3 may represent a target for lithium's mood-stabilizing properties. Specifically, it has been demonstrated that lithium administration regulates multiple GSK-3 targets in vivo and that multiple additional classes of mood-stabilizing and antidepressant drugs regulate GSK-3 signaling. Pharmacological or genetic inhibition of GSK-3 results in mood stabilizer-like behavior in rodent models, and genetic association studies implicate GSK-3 as a possible modulator of particular aspects of bipolar disorder including response to lithium. Furthermore, numerous recent studies have provided a more complete understanding of GSK-3's role in diverse neurological processes strengthening the hypothesis that GSK-3 may represent a therapeutically relevant target of lithium. For example, GSK-3 is a primary regulator of neuronal survival, and cellular responses to glucocorticoids and estrogen may involve GSK 3-regulated pathways. While the preclinical evidence discussed in this review is encouraging, ultimate validation of GSK-3 as a therapeutically relevant target will require clinical trials of selective novel inhibitors. In this regard, as is discussed, there is a major effort underway to develop novel, specific, GSK-3 inhibitors. PMID- 15827568 TI - Enhanced acquisition of cocaine self-administration in rats developmentally exposed to lead. AB - The rate of acquisition of drug self-administration may serve as a predictor of later drug-taking behavior, possibly influencing the vulnerability to use drugs. The present study examined the effects of perinatal (gestation/lactation) lead exposure on adult rates of acquisition of intravenous cocaine self-administration using an automated procedure that included both Pavlovian and operant components. For Experiment 1, female rats were gavaged daily with 0 or 16 mg lead for 30 days prior to breeding with nonexposed males. Metal administration continued through pregnancy and lactation and was discontinued at weaning (postnatal day (PND) 21). Animals born to control or lead-exposed dams subsequently were tested daily as adults in a preparation where sessions included an initial 3-h autoshaping period followed by a 3-h self-administration period where 0.20 mg/kg cocaine was delivered contingently. During autoshaping, intravenous cocaine infusions were paired with the extension and retraction of a lever, while infusions occurred during self-administration only when a lever press was executed (FR-1). The criterion for acquisition was a 2-day period during which a mean of 50 infusions/session occurred during self-administration. Animals were given 35 days to reach criterion. In Experiment 1, accelerated rates of acquisition of cocaine self-administration were evident for lead-exposed animals relative to controls. Overall, the number of self-administered cocaine infusions per session was significantly higher for lead-exposed rats as compared to control rats. Experiment 2 replicated Experiment 1 except that a higher dose of cocaine (0.80 mg/kg) was employed as the reinforcer, and 30 infusions/session was the set criterion. At the higher cocaine dose (Experiment 2), acquisition rates for control and lead-exposed animals were not markedly different, and significantly different infusion rates were not observed. PMID- 15827569 TI - The effect of citalopram pretreatment on neuronal responses to neuropsychological tasks in normal volunteers: an FMRI study. AB - Changes in serotonin neurotransmission have also been implicated in the etiology and treatment of impulse control disorders, depression, and anxiety. We have investigated the effect of enhancing serotonin function on fundamental brain processes that we have proposed are abnormal in these disorders. In all, 12 male volunteers received citalopram (7.5 mg intravenously) and placebo pretreatment in a single-blind crossover design before undertaking Go/No-go, Loss/No-loss, and covert (aversive) face emotion recognition tasks during functional magnetic resonance imaging (fMRI). Blood oxygenation level dependent responses were analyzed using Statistical Parametric Mapping (SPM2). The tasks activated prefrontal and subcortical regions generally consistent with literature with lateral orbitofrontal cortex (BA47) common to the three tasks. Citalopram pretreatment enhanced the right BA47 responses to the No-go condition, but attenuated this response to aversive faces. Attenuations were seen following citalopram in the medial orbitofrontal (BA11) responses to the No-go and No-loss (ie relative reward compared with Loss) conditions. The right amygdala response to aversive faces was attenuated by citalopram. These results support the involvement of serotonin in modulating basic processes involved in psychiatric disorders but argue for a process-specific, rather than general effect. The technique of combining drug challenge with fMRI (pharmacoMRI) has promise for investigating human psychiatric disorders. PMID- 15827570 TI - Arterial pressure and heart rate increase during REM sleep in adenosine A2A receptor knockout mice, but not in wild-type mice. AB - Rapid eye movement (REM)-sleep related changes in arterial pressure (AP) and heart rate (HR) were observed in homozygous and heterozygous adenosine A(2A) receptor (A2AR) knockout (KO) mice, and the corresponding wild-type mice. During REM sleep, the mean AP (MAP) and HR were clearly increased in the homozygous A2AR KO mice, while, in the wild-type mice, they were decreased or maintained at the same level. Neither homozygous nor heterozygous A2AR KO mice showed significant difference in diurnal pattern and the hourly values of MAP and HR compared to the wild-type mice. From these findings, it is likely that the adenosine A2AR is involved in autonomic regulation during REM sleep. PMID- 15827571 TI - The RGSZ2 protein exists in a complex with mu-opioid receptors and regulates the desensitizing capacity of Gz proteins. AB - The regulator of G-protein signaling RGS17(Z2) is a member of the RGS-Rz subfamily of GTPase-activating proteins (GAP) that efficiently deactivate GalphazGTP subunits. We have found that in the central nervous system (CNS), the levels of RGSZ2 mRNA and protein are elevated in the hypothalamus, midbrain, and pons-medulla, and that RGSZ2 is glycosylated in synaptosomal membranes isolated from CNS tissue. In analyzing the function of RGSZ2 in the CNS, we found that when the expression of RGSZ2 was impaired, the antinociceptive response to morphine and [D-Ala2, N-MePhe4, Gly-ol5]-enkephalin (DAMGO) augmented. This potentiation involved mu-opioid receptors and increased tolerance to further doses of these agonists administered 24 h later. High doses of morphine promoted agonist desensitization even within the analgesia time-course, a phenomenon that appears to be related to the great capacity of morphine to activate Gz proteins. In contrast, the knockdown of RGSZ2 proteins did not affect the activity of delta receptor agonists, [D-Pen2,5]-enkephalin (DPDPE), and [D-Ala2] deltorphin II. In membranes from periaqueductal gray matter (PAG), both RGSZ2 and the related RGS20(Z1) co-precipitated with mu-opioid receptors. While a morphine challenge reduced the association of Gi/o/z with mu receptors, it increased their association with the RGSZ2 and RGSZ1 proteins. However, only Galphaz subunits co precipitated with RGSZ2. Doses of morphine that produced acute tolerance maintained the association of Galpha subunits with RGSZ proteins even after the analgesic effects had ceased. These results indicate that both RGSZ1 and RGSZ2 proteins influence mu receptor signaling by sequestering Galpha subunits, therefore behaving as effector antagonists. PMID- 15827572 TI - Effect of increased serotonin levels on [18F]MPPF binding in rat brain: fenfluramine vs the combination of citalopram and ketanserin. AB - [18F]MPPF is a selective serotonin-1A (5-HT1A) receptor antagonist and may be used to measure changes in the functional levels of serotonin (5-HT). The technique is based on the assumption that the injected radiolabeled ligand competes for the same receptor as the endogenous transmitter. Results from studies using serotonergic ligands are not always consistent. The aim of the present study was to investigate if [18F]MPPF binding is decreased after an increase in 5-HT levels. [18F]MPPF binding was assessed in conscious rats using ex vivo autoradiography. We studied the effect of the 5-HT-releasing agent and reuptake inhibitor fenfluramine (10 mg/kg i.p.) and of a combination of the selective serotonin reuptake inhibitor (SSRI) citalopram (10 micromol/kg, s.c.) with the 5-HT2C antagonist ketanserin (100 nmol/kg, s.c). The effect of both treatments on extracellular 5-HT levels was determined using microdialysis. Fenfluramine treatment resulted in a 30-fold increase in extracellular 5-HT levels in the ventral hippocampus and induced a significant reduction of [18F]MPPF binding in the frontal cortex, hypothalamus, amygdala, and hippocampus. The microdialysis results showed a 10-fold 5-HT increase in the ventral hippocampus after combined administration of ketanserin and citalopram. The combination, however, did not affect [18F]MPPF binding. Our data show that [18F]MPPF binding in conscious rats is only reduced after substantial and therefore nonphysiological increases in 5-HT levels. These results may imply that the majority of 5-HT1A receptors is in the low-affinity state, in vivo. PMID- 15827573 TI - Dopamine transporter genotype and methylphenidate dose response in children with ADHD. AB - Stimulant medications, such as methylphenidate (MPH), are the most commonly used, effective treatment for ADHD. MPH acts primarily by inhibiting the dopamine transporter (DAT), a protein responsible for the reuptake of dopamine from the synapse into presynaptic terminals. We sought to evaluate the relationship between DAT1 3'-untranslated region (3'-UTR) variable number tandem repeats (VNTR) genotypes and dose response to MPH. Children with ADHD (n=47), ages 5-16 years (mean=9.02 years), underwent a 4-week, double-blinded, crossover trial with forced weekly dosage changes. Children were genotyped for the DAT1 VNTR and evaluated on placebo and three dosage levels of OROS MPH. Parents and clinicians who were blind to genotype and medication status rated ADHD symptoms, impairment, and stimulant side effects each week. Children who were homozygous for the less common, 9-repeat DAT1 3'-UTR genotype displayed a distinct dose-response curve from that of the other genotype groups, with an absence of typical linear improvement when the dose was increased from 18 mg to 36 and 54 mg. Further research is needed to determine the mechanisms related to poor response in patients with the 9/9-repeat genotype, and to determine if this group responds differentially to alternative treatments. PMID- 15827575 TI - [Clinical trials, pharmaceutical companies and clinical practice]. PMID- 15827574 TI - Prenatal disruption of neocortical development alters prefrontal cortical neuron responses to dopamine in adult rats. AB - A growing body of evidence suggests that structural changes in the cortex may disrupt dopaminergic transmission in circuits involving the prefrontal cortex (PFC) and may contribute to the etiology of schizophrenia. In this study, we utilize a rodent model of neonatal disruption of cortical development using prenatal administration of the mitotoxin methylazoxymethanol acetate (MAM). Using intracellular recordings in vivo, we compare the physiology of prefrontal cortical neurons and their responses to topical administration of dopamine (DA) in intact animals and adult rats treated prenatally with MAM. Topical administration of DA hyperpolarized the membrane potential (MP) and decreased the firing rate of neurons recorded in deep layers of the PFC in intact animals. Furthermore, electrical stimulation of the VTA evoked fast onset epsps or long lasting depolarizations in PFC neurons. In comparison, PFC neurons recorded in MAM-treated animals had significantly faster baseline firing rates. Moreover, topical administration of DA did not affect the MP or firing rate of the neurons in MAM-treated animals. However, MAM-treated animals exhibited an increase in the percentage of neurons responding with long-lasting depolarizations to stimulation of the VTA. The results of this study indicate that PFC neurons in the MAM treated rats are not responsive to DA administered superficially, while at the same time exhibit greater responsiveness to VTA stimulation. These results are consistent with a rewiring of the corticolimbic system in response to neurodevelopmental insults. PMID- 15827576 TI - [Gene therapy: what is it and what is its use?]. AB - Gene therapy has developed as a method of approach to the treatment of human diseases based on the transfer of genetic material to the cells of an individual. Normally, the aim of this transfer of genetic material is to re-establish a cellular function that has been abolished or is defective, to introduce a new function or to interfere with an existing function. Thus, the different gene therapy strategies are based on the combination of three key elements: the genetic material to be transferred, the method of transfer and the cellular type that will incorporate this genetic material. Attention was initially centred on the treatment of monogenic hereditary diseases, but subsequently the majority of clinical trials (over four hundred) have concerned the treatment of cancer. In China a genetic product has been approved for commercialisation: an adenovirus that transfers the correct version of the tumour suppressor gene p53. And, in the late 1990s, a group of children with severe combined immunodeficiency were successfully treated through the ex vivo transfer of the correct version of the altered gene to their bone marrow, although some of these children later developed lymphoproliferative syndromes due to the activation of an oncogen in the corrected cells. Human gene therapy is feasible and can be useful, but the tools need improving for it to become part of the therapeutic arsenal. PMID- 15827578 TI - [Cognitive state as a conditioner of frailty in the elderly. Perspective from a health centre]. AB - AIM: To determine the influence of the cognitive state on the presence of different frailty factors in the elderly. METHODS: Study of an outpatient elderly population with chronic diseases (resident at home or institutionalised), the presence of different frailty risk factors and their relation to cognitive state (measured using the mini-mental state examination-MEC). RESULTS: Study of 147 elderly people with an average age of 71.4 years and a similar proportion of men (74; 50.3%) and women (73; 49.7%). Thirty-four subjects (23.1%) institutionalised in residences. The percentage of patients showing cognitive impairment (MEC<24 points) is 12.9% (19 cases). Presence of frailty risk factors: low social support: 7.5% (11); falls: 17% (25); urinary incontinence: 18.4% (27); depression: 13.6% (20); anxiety-insomnia: 29.9% (44); hospitalisation readmissions: 21.8% (32); multiple medications (>3 medicines): 53.7% (79); pluripathology (> or =3 diseases): 36.1% (53). The frailty factors that have a significant relation in patients with cognitive impairment (MEC<24) are falls [OR=59.5 (CI 95%=14.7-240.6)] (p<0.0001), urinary incontinence [OR=31.2 (8.9 109.1)] (p<0.0001), hospitalisation-readmissions [OR=32.9 (8.6-125.8)] (p<0.0001) and depression [OR=7.8 (2.5-23.5)] (p<0.0001). With respect to scoring on the MEC by percentiles, the risk factors that showed a tendency of lineal appearance are falls (p<0.0001), urinary incontinence (p<0.0001), hospitalisation-readmissions (p<0.0001) and pluripathology (p=0.002). CONCLUSIONS: Cognitive impairment marks the appearance in a significant form of frailty factors in the elderly, such as falls, urinary incontinence, hospitalisation-readmissions and depression. This relation is not only appreciable in patients with an established cognitive impairment (MEC<24 points), but there is also a trend to appear as this impairment progresses, with a statistical relation for falls, urinary incontinence, hospitalisation-readmissions and pluripathology. PMID- 15827577 TI - [Study of the resistance of Mycobacterium tuberculosis to the principal tuberculostatics (Hospital of Navarra 1996-2003)]. AB - INTRODUCTION: The aim of this study was to evaluate the sensitivity to the principal tuberculostatics of the M. tuberculosis stocks isolated in our laboratory and to study the factors related to resistance. METHODS: Study of 475 stocks of M. tuberculosis corresponding to all cases diagnosed in the eight year period between 1996 and 2003. We employed the BACTEC 460TB system, together with Lowenstein solid medium, in the cultivation of the samples. The sensitivity studies were carried out using the BACTEC 460TB system. RESULTS AND CONCLUSIONS: Both the incidence and the number of cases of resistant tuberculosis showed oscillations over the eight years of the study. The fact that there is no homogeneous tendency makes it necessary to maintain active surveillance of this process. Global resistance to isoniacide was 8%, making it is convenient to carry out sensitivity studies in all the diagnosed cases. The cases of multiresistant tuberculosis (resistant to at least isoniacide and rifampicin), did not exceed 3%. The prognosis of the patients with resistant tuberculosis was bleak in cases of coinfection with HIV; however, when the immunological defences were conserved, and guidelines for treatment with active drugs were provided, the recovery of the majority of the patients was achieved. At present, sensitive and rapid procedures are available to us, making it recommendable to study the sensitivity of all the stocks of M. tuberculosis that are isolated; this is essential in the case of HIV positive or immigrant patients. PMID- 15827579 TI - [Cardiac magnetic resonance: clinical applications]. AB - Cardiovascular disease is the most frequent cause of mortality in the developed countries and represents a serious social, economic and health problem. Although very diverse, useful techniques exist for diagnosing cardiac diseases, it is frequently necessary to ask for more than one test to reach a specific diagnosis. Magnetic resonance is a harmless, well tolerated and safe technique, which is currently available in the majority of hospitals. This technique makes it possible in a single exploration to study the anatomy of the heart and to make a qualitative, semi-quantitative and quantitative assessment of the parameters of cardiac function. It provides information of cardiac and vascular anatomy and function in complex congenital cardiopathies. Besides, with the administration of intravenous contrast, it enables knowledge to be gained of myocardial viability in ischaemic cardiopathy. Hence, cardiac magnetic resonance is emerging as one of the most promising techniques for the study of congenital and acquired cardiac pathology. PMID- 15827580 TI - [Infections in non-transplanted oncohaematological patients]. AB - Oncohaematological patients present a high incidence of infections, which are one of the principle causes of morbidity and mortality. There are different types of immunodepression related to the disease, the moment of its evolution and the treatment received. For practical purposes we will distinguish between patients with severe neutropenia, those with some alteration to humoral immunity and, finally, cellular immunodeficiencies. There are no immunodeficiencies associated to each disease, instead several immunitarian deficiencies can be associated in a single clinical entity. Neutropenic patients, generally with acute leukaemias and following intensive chemotherapy, have bacterial and fungal infections conditioned by the intensity and duration of the neutropenia. In the case of patients with humoral immunodeficiency (multiple myeloma, chronic lymphatic leukaemia, splenectomised) there are frequent infections by encapsulated germs. When there is cellular immunodepression (Hodgkin's disease, advanced chronic lymphoproliferative syndromes, treatment with glucocorticoids, analogues of the purines and treatment with monoclonal antibodies) the risk of infection by opportunist germs is conditioned by the reduction of the figure of CD4 lymphocytes. We review the different strategies of prophylaxis and treatment in each of the situations. PMID- 15827581 TI - [Occupational disability due to locomotor pathology in the Foral Community of Navarra. Epidemiological aspects]. AB - INTRODUCTION: Disability produced by locomotor pathology causes a loss in working hours and indirect costs that are increasing at a growing rate. The aims of this study are to estimate the costs in total working hours lost caused by this pathology in two health areas of Navarra with 441,600 inhabitants, to obtain standards for the duration and incidence of sick leave and to study the relation between the average duration of sick leave and variables such as the geographical accessibility of the rehabilitation services, residence in a rural or urban milieu, and between the different health areas. METHODS: We study all the processes of temporary disability due to pathology of the locomotor system in Health Areas I and III of Navarra in 1997 and 1998, taking different variables into account. We make a descriptive study of the pathologies, counting the working days lost. We calculate the annual incidence of sick leave because of pathologies. We compare average duration with other variables. RESULTS: These represent 221,054 lost working days per year. Low back pain and sprained ankles are the most frequent processes. We find an increase in the incidence of temporary disability due to low back pain and lumbar/sciatica in the basic rural areas of the middle region. There are no statistically significant differences with a p<0.05 in the duration of temporary disability according to Health Area, rural or urban milieu or geographical accessibility to the Rehabilitation Services. A statistically significant fall can be observed in the first year of operation of a temporary disability management program. PMID- 15827582 TI - [Communicable disease surveillance in Navarre, 2004]. AB - Epidemiological surveillance in Navarre (584,734 inhabitants) covers 34 transmissible diseases, whose notification is compulsory, and epidemic outbreaks of any aetiology. Notification is carried out on a weekly basis by the doctors from paediatrics, primary care and specialised care. In 2004, 75.8% of all the possible notification reports (a weekly report for each doctor) were received, a percentage that has improved in the last five year period. Flu only reached 14.4 cases per 1,000 inhabitants (Epidemic Index, EI: 0.30), due to the advance of the epidemic peak for the 2003-2004 season to the month of November. The rate of respiratory tuberculosis fell to 11.6 cases per 100,000 inhabitants, and the rate of non-respiratory tuberculosis rose to 2.7 per 100,000. Ten cases of tuberculosis (11.9%) were grouped into four outbreaks that affected adolescents and young adults. Thirty percent of the cases were produced in immigrants and 4.8% in persons coinfected with HIV, proportions that are similar to those of the previous year. Eleven cases of meningococcal disease were reported, (1.9 cases per 100,000 inhabitants; EI 0.73), but only in 8 cases was the clinical form sepsis and/or meningitis. Neisseria meningitidis serogroup B was isolated in 8 cases, and serogroup C in 2 cases, the latter 2 were adults and were not vaccinated. The incidence of immunopreventable diseases continues to fall, and for the fifth consecutive year no case of measles has been reported. Legionnaire's disease, which is detected through the systematic determination of the antigen in urine, rose to 5.8 cases per 100,000 inhabitants (EI: 1.42), without any epidemiological relation between them. The incidence of imported diseases rose, with 12 cases of malaria, 8 of shigellosis, 5 of hepatitis A and 2 of legionnaire's disease acquired outside Spain. PMID- 15827583 TI - [Will methyldibromo glutaronitrile (Euxyl K400) be the contact allergen of the year 2005 in Navarra?]. PMID- 15827584 TI - [Duodenal gangliocytic paraganglioma]. AB - We present the case of an 85 year old male who was admitted to hospital with abdominal pain and jaundice. Different explorations were performed for this reason, with a mass observed in his 2nd duodenal portion. Histological study showed that it was a duodenal gangliocytic paraganglioma. The clinical characteristics of this infrequent tumour are described and we review the diagnosis and treatment. PMID- 15827585 TI - [Intestinal invagination in the adult secondary to jejunal lipoma]. AB - Small bowel intussusception is an unusual pathology in the adult. Most commonly, it is secondary to intestinal wall organic disorders. A complete small bowel obstruction is the most frequent clinical presentation, which requires emergency surgery in many cases. The preoperative diagnosis is infrequent. The best treatment is surgical resection. PMID- 15827586 TI - [Psychosis associated to megacisterna magna]. AB - Megacisterna magna is a developmental malformation of the middle-line brain structures, which exists along a continuum with cerebellum hypoplasia (Dandy Walker variant) and cerebellum agenesia (Dandy-Walker syndrome). Psychotic disorders, and particularly schizophrenia, are now conceptualised as disorders of neurodevelopment. We report on a patient who presented a psychotic disorder (delusional type), and mega cisterna magna. This is the first report of such an association, and it is suggested that both the psychotic disorder and the mega cisterna magna may be the expression of a single underlying neurodevelopment abnormality. PMID- 15827587 TI - The effect of training on the reporting of notifiable diseases among health workers in Yobe State, Nigeria. AB - The generation of data through disease surveillance and notification system is critical to appropriate planning and implementation of disease control programmes, outbreak investigation, emergency preparedness and response. Health workers therefore need to be trained, retrained and updated on the principles and practice of disease surveillance and notification. This quasi-experimental study compared a study and control group "before and after" an intervention (training programme) in the study group. The Experimental and control LGA's were selected using a multistage, stratified random sampling technique. Overall, three LGA's were selected and enrolled in each of the groups. In each of the selected LGA's, all functional health facilities and personnel that fulfilled the inclusion criteria were then included in the study. The total number of participants in the experimental and control groups were 73 and 71 respectively at baseline. The proportion of personnel who were aware of the surveillance system increased from 35.6% to 91.9% (p=0.00) and the mean knowledge score increased from 0.85+/-1.38SD to 6.152.64SD (p=0.00) post intervention in the experimental group. The percentage completeness was 2.3% before and 52.0% after (p-0.00), while the percentage timeliness was 0.0% before and 42.9% after (p=0.00) in the experimental group. These statistically significant differences were however not demonstrated in the control group. Training therefore had a positive effect on health personnel knowledge, reporting requirement and the timeliness and completeness of the disease surveillance and notification system. PMID- 15827588 TI - Arteriovenous fistula for patients on long term haemodialysis in jos, Nigeria. AB - BACKGROUND: The introduction of haemodialysis has prolonged the lives of patients with end-stage-renal disease (ESRD). To maintain them on long-term dialysis, vascular access procedures are required. OBJECTIVES: To describe the experience with arteriovenous fistula (AVF) from a developing country. METHODS: Eighty-two AVF were created in 74 patients on maintenance haemodialysis. They all had side to-end AVF using prolene 7/0 monofilament suture for anastomoses. RESULTS: There were 55 males and 19 females with a male: female ratio of 2.9:1. The ages ranged from 18 to 70 years with a mean of 43.4 "12.1 years. Most of the AVF created were on the left upper limbs with the radio-cephalic and brachio-cephalic accounting for more than 70% . Eight patients developed thrombosis of the veins used for the AVF and one developed a pseudo aneurysm. CONCLUSIONS: Arteriovenous fistula makes long-term haemodialysis feasible. It can be created easily and has a low rate of complications. PMID- 15827589 TI - Effect of acute uncomplicated malaria on platelet counts. AB - OBJECTIVE: To determine the effect of uncomplicated malaria on platelet count. PATIENTS AND METHODS: Adult Nigerians (residing in the Lagos metropolis) who were diagnosed as having uncomplicated malaria were treated with a standard regime of chloroquine. Platelet counts were obtained before treatment, Day 0, and again on day 14 when parasitaemia had been cleared in the patients. There were 40 (21 males, 19 females) patients enrolled for the study. Their ages ranged from 15 years to 56 years with a mean +/- SEM of 27. 4 +/- 1.8 years. 28 (14 males, 14 females) patients had both day 0 and 14 platelet counts. RESULTS: The malaria parasite counts ranged from 1020 /mm(3) to 72,000 /mm(3) at day 0 with a mean +/ SEM of 15,638.0 +/- 3,727.0/mm(3) and zero on day 14. The mean platelet count on day 0 was 137.0 +/- 58.4x 10(9)/L while the day 14 mean platelet count was 234.0 +/- 96.9 x 10(9)/L. Comparison of mean platelet counts on day 0 with those on day 14 showed a highly statistically significant difference (p < 0.001). The degree of malaria parasitaemia was not significantly related to the level of platelet count. Transient thrombocytopenia is very common in uncomplicated malaria in semi -immune adults. The mechanism, aetiology, and clinical relevance of the phenomenon deserve further studies. PMID- 15827590 TI - Basic trauma life support: knowledge of medical students. AB - BACKGROUND: The first hour of management (golden hour) of any trauma patient might be the determining factor if he/she will survive. The first contact most patients have in an emergency room is with a young medical graduate. The knowledge of basic trauma life support (BTLS) by all doctors therefore becomes important. AIMS AND OBJECTIVES: This study set out to assess how much knowledge medical students had about BTLS in their final year compared with their knowledge at the beginning of clinical postings. METHOD: A cross-sectional study of fourth and sixth (final) year medical students to evaluate their knowledge of BTLS using a test of 30 questions under examination conditions. One hundred and twenty-three fourth year students were given a test after a lecture on the care of injured patients. The same test was administered on 41 sixth year students without prior lecture on day of test, having had a similar lecture 2 years before. Data analysis was performed using the SPSS 11.0 for Windows statistical software and EPI INFO version 6.04. RESULT: Mean scores were 25.8 +/- SD1.91 for the 4th year students and 21.0 +/- SD2.93 for the 6th year students. There was a statistically significant difference between scores of the 4th year and the 6th year (P < 0.000001). CONCLUSION: Knowledge was imparted in the 4th year but was forgotten by the 6th year. It is essential that courses in BTLS be given periodically to all medical graduates and all personnel involved in care of the traumatised patient. PMID- 15827591 TI - Gene therapy in the management of oral cancer: a review of recent developments. AB - AIMS: This article reviews the present body of knowledge regarding the principle, transfer techniques, therapeutic strategies, clinical applications and limitations of gene therapy in the management of oral squamous cell carcinoma. MATERIALS AND METHODS: Scientific publications on gene therapy between 1990 and 2003 were selected for the purpose of the review. These include clinical articles, experimental studies and review articles. RESULTS: Viruses are the commonly used transfer system for the delivery of gene therapy. The viral vectors commonly used are: retroviruses, adenoviruses, herpesviruses, and adenoassociated viruses (AAV). Transfection of cancer cells in vivo with gene therapy is done by intralesional injection, and sometimes by topical application. Phase I and II clinical trials have established the safety and clinical efficacy of gene therapy in the treatment of oral squamous cell carcinomas in humans, especially in combination with chemotherapy or radiotherapy. Phase III clinical trials and studies of the use of gene therapy as an adjuvant following surgery are presently underway. CONCLUSIONS: Gene therapy represents a new and innovative approach to the treatment of oral cancer especially in recurrent disease and adjuvant treatment. Oral squamous cell carcinoma is especially an attractive tumour target due to its frequent genetic mutations and accessibility for intra-tumoural administration of gene therapy. However, gene therapy has not yet been shown to be suitable for systemic delivery in cancer patients, hence the control of regional and metastasis is presently difficult. PMID- 15827592 TI - Helminthiasis in selected children seen at the University of Benin Teaching Hospital (UBTH), Benin City, Nigeria. AB - OBJECTIVES: Illiteracy, poverty with associated poor environmental sanitation practices have been implicated in the heavy burden of helminthiasis among children. The objective of this cross-sectional survey is to determine the impact of parents' level of education on the intestinal helminthic status of children. PATIENTS, MATERIALS AND METHODS: All patients, 0-15 years totaling 1030 who visited the communicable disease clinic of the University of Benin Teaching Hospital, Benin City, from October, 2001 to March 2002 were included in this study. However, only 207 of the enlisted patients submitted faecal samples for examination and constituted the study population. Tool for data collection was structured questionnaire that was researcher administered to mother/caregivers after informed consent was obtained. Information sought in the questionnaire included sex and age of the subjects and level of education of their parents. Formol ether concentration method was used for stool examination while intensity was estimated using McMaster counting chamber technique. RESULTS: Of the 207 faecal samples examined 46(22.2% ) had ova of helminths while 161(77.85% ) had none. More school children, 40(19.3% ), than pre-school children, 6(2.9% ) had helminth ova in their faecal samples and the difference was found to be statistically significant (P < 0.05). More females, 28(13.5% ) than males, 18(8.7% ) were infected but the difference was however not statistically significant (P > 0.05). Intestinal helminth ova seen in order of prevalence include A. lumbricoides, 23(11.1% ); hookworm, 12(5.8% ); T. trichiuria, 8(3.8% ); Schistosoma mansoni, 2(1.0% ); Strongyloides stercoralis, 1(0.5% ) while multiple infection was recorded in 11(5.3% ) patients. Intensity of infection was low. The difference between the means for A. lumbricoides, T. trichiuria and multiple infection by age were found to be statistically significant P < 0.001; P < 0.05; P < 0.001, respectively. Mother's level of education more than father's level of education seemed to influence the pattern of intestinal helminthiasis among the children but the association was not found to be statistically significant (P > 0.05). CONCLUSION: Prevalence and intensity of infection was low probably due to periodic antihelminthic treatment offered by the health authorities. Improvement in mother's level of education brought about decrease in prevalence of intestinal helminthiasis among the children. Therefore, improvement in female education should be encouraged to reduce the incidence of communicable disease in the family. PMID- 15827593 TI - Childhood parapneumonic pleural effusion in Enugu. AB - OBJECTIVES: To review the epidemiological pattern and clinical features, management and outcome of childhood parapneumonic pleural effusion in Enugu. METHODS: The records of the children with diagnosis of pleural effusion admitted at the University of Nigeria Teaching Hospital, Enugu between January 1995 and February 2001 were reviewed. RESULTS: Forty out of forty two patients reviewed were analysed. There were 20 males, and 20 females (ratio of 1: 1), with age range of 3 months to 156 months (13 years) and the mean age of 4.99 years. The presenting symptoms were fever in 40 patients (100% ), cough in 38 (97.40% ), dyspnea in 36 (90.00% ), breathlessness in 27 (67.5% ), while, the most common physical sign was pallor. Thirty six children had diagnostic thoracocentesis. Twenty four (55.81% ) yielded pus, out of which, 10 grew organisms on culture; Streptococcus Pneumonia in 3, Staphylococcus aureus in 2, coliforms in 2, Haemophilus influenzae in 1, Proteus mirabilis in 1, Pseudomonas aeruginosa in 1. Nine patients received ampicillin/cloxacillin combination alone; 31 received ampicillin/cloxacillin with other antibiotics, including anti- tuberculous drugs in four children. Thirty-five of the children had thoracotomy, and one later had thoracotomy and decortication. Thirty-seven (92.5% ) were discharged home on recovery. The average admission duration was 24 days. Mortality rate was 7.50%. CONCLUSIONS: In this review, there appears to be an evolving change in epidemiological pattern in comparison with the previous study from this center. There is need to improve on the diagnostic procedure and treatment modalities in order to establish definite criteria for antibiotic use and surgical intervention. PMID- 15827594 TI - A review of the radiological diagnosis of small bowel obstruction using various imaging modalities. AB - Radiological investigation is an important tool in the diagnosis of small bowel obstruction (SBO). While plain abdominal radiograph remains the commonest investigation done in SBO, contrast gastrointestinal radiography, ultrasonography (US), computerised tomography (CT) scan -- with or without contrast enhancement are increasingly being advocated particularly in equivocal cases of SBO. We conducted manual, medicine and internet search for relevant literature on diagnosis of SBO using radiological investigations. Plain abdominal radiograph has a diagnostic accuracy for SBO that varies widely from 50 - 92% . Contrast gastrointestinal radiograph in equivocal cases of adhesive SBO can readily differentiate between complete and partial obstruction. Abdominal ultrasound is cheap and readily available. Accuracy of as high as 89% has been reported in experienced hands. CT scan has an added advantage of predicting the site and cause of obstruction. In developing countries like ours, plain abdominal radiography still remains an important diagnostic tool. The role of abdominal ultrasonography needs further evaluation since it is safe, readily available and affordable. PMID- 15827595 TI - Bile duct stricture -- a sign of things to come? AB - OBJECTIVE: The aim of the report is to highlight this difficult to-treat condition hitherto uncommon in our environment and warn of the likelihood of more to come with the rising incidence of gallstone in our sub region. PATIENT AND METHOD: A case report of an 18-year-old male undergraduate patient. RESULT: A successful management with clinical, biochemical and radiological evidence (OTC and tubograms) despite inadequate investigative tool. PRINCIPAL CONCLUSION: With the rising incidence of gall stones in our environment, it is imperative that the Trainee Surgeon must be schooled in common biliary surgery and help prevent this dreaded complication (bile duct stricture) and other sequelae of gall bladder surgery. A plea is also made for the provision of essential investigative tool to facilitate management of such cases in recognised centers. PMID- 15827596 TI - Permanent hypoparathyroidism developing post-thyroidectomy. AB - A case of permanent hypocalcaemia following a subtotal thyroidectomy for a simple multinodular goiter in a 35 year -- old married teacher is presented. This further confirms the need for utmost precaution during thyroid surgery to prevent the damage or inadvertent removal of the parathyroid glands and its consequential complications. It also throws more light on the problem faced in the management of hypocalcaemia vis a vis patient compliance and availability of proper medications in this environment. PMID- 15827597 TI - Extramedullary presentation of multiple myeloma in the parotid gland as first evidence of the disease -- a review with case report. AB - Myeloma is a neoplasm of plasma cells that causes painful, bone-destructive lesions. It may present as a solitary lesion (plasmacytoma), or involve multiple sites (multiple myeloma). Myeloma may occur in an extraosseous location as a solitary lesion, or less commonly, in combination with multiple myeloma. Oral manifestations may be the first sign or symptom of the disease. Soft tissue plasmacytomas of the head and neck tend to involve the nasal cavity or nasopharynx, rather than the oral cavity. Its occurrence in the parotid gland is rare. We present an unusual case of multiple myeloma with extramedullary involvement involving the parotid gland as first evidence of the disease. Multiple myeloma should be considered in the differential diagnosis of salivary gland swellings. PMID- 15827598 TI - Neurofibromatosis with malignant transformation presenting as an emergency in a nigerian child: a case report. AB - Neurofibromatosis is a rare disease, and malignant transformation of this otherwise benign tumor is extremely rare in childhood. In this case report, attention is drawn to childhood neurofibromatosis presenting as a life threatening triad of malignant transformation, visceral metastasis and upper airway obstruction leading to death in a 9-year-old female Nigerian child. This case report is purposed to draw attention to this condition, mostly reported in adults and to review literature on the disease. PMID- 15827599 TI - Destructive ophthalmic surgical procedures in Onitsha, Nigeria. AB - OBJECTIVES: to determine the incidence, trend and causes for surgical removal of the eyeball at the Guinness Eye Center Onitsha, Nigeria. PATIENTS AND METHODS: Case files and theatre records of all patients who had eviscreration, enucleation or exenteration at the Guinness Eye Center Onitsha between 1995 and 2001 were reviewed. Information on age, sex, disease duration before hospital presentation, indication for surgery and the type of surgery were analysed. RESULTS: 56 eyes of 56 patients were surgically removed. Patients including some with penetrating eye injuries presented late for treatment; 37.5% and 53.5% had used traditional eye medicines (TEM) and eye drops possibly containing steroids, respectively, prior to hospital consultation. The indications for surgery were severe ocular infections, especially panophthalmitis; severe penetrating eye injuries and orbito-ocular tumours. The annual incidence ranged from 1.9 - 4.2% with an average of 3.4% . The downward trend observed in the later part of the study was associated with reduced incidence of ocular gunshot injuries, which in turn coincided with a reduction in violent crime in Onitsha and its environs. CONCLUSIONS: In spite of the observed downward trend in the rate of destructive eye surgery, the incidence of such procedures could be reduced further by timely intervention with modern microsurgical techniques; intraocular antibiotics and radiotherapy. The public should be educated on the harmful effects of TEM and abuse of eye drops. Those with eye problems should seek early hospital treatment. PMID- 15827600 TI - Limb gangrene following treatment of limb injury by traditional bone setter (Tbs): a report of 15 consecutive cases. AB - This study is done to evaluate cases of limb gangrene resulting from treatment of limb injury by traditional bonesetter. METHOD: This is a prospective study in which patients with limb gangrene are evaluated as each presents. Data extracted for evaluation include history of having been to a traditional bonesetter, the original injury, type of treatment given by the traditional healer, progression of condition while on treatment and reason for not coming to orthodox orthopaedic centre ab initio for treatment. RESULTS: 15 cases were seen during the 5-year of study spanning June 1997 to May 2002. Upper limbs were involved in 4 cases while 11 involved the lower limbs. The original injuries were 3 cases of soft tissue injury to the joints, 7 closed fractures and 5 open fractures of type I and II. There was associated sepsis and toxemia in all except one. All the cases were treated by amputation. There were 4 deaths. CONCLUSION: Limb gangrene was not a justifiable end-result of treatment in all the cases judging by the nature of the original injury. Reasons for this end result were adduced. The need to avert unnecessary limb loss from mismanagement of limb injury by education of public in general and enlightenment of the traditional bonesetters of the harmful procedures in their practice is highlighted. PMID- 15827601 TI - Zygomatic arch reconstruction with autogenous rib bone graft in a post irradiated patient -- a case report. AB - Reconstruction of zygomatic arch using an autogenous free non-vascularised bone graft is presented. The zygomatic arch was resected along with the masseter and part of temporalis muscle following a histologically diagnosed case of chondrosarcoma. She received radiotherapy 3 weeks post-operatively to the site. The resultant facial asymmetry was unacceptable to the patient. She was therefore rescheduled for reconstruction of the zygomatic arch despite the anticipated attendant risk of avascular bed for the graft. Bone grafting was done 44 months post irradiation and the patient had been followed up for 38 months without evidence of rejection. We evaluated other sites of possible bone of similar contour and thickness to the zygomatic arch; we found the rib most suitable, despite the poor quality of rib as graft material, because it is a dense, thick bicortical bone with low proportion of cancellous bone, although it is the second most favoured donor site for free bone graft. The advantages of the rib as a graft are that it presents a contour that could be shaped to fit many defects, it is available in sufficient length up to 12-18 cm to fit large defects, the donor site regenerates when the periosteum is preserved. PMID- 15827602 TI - Death from ingestion of removable partial denture: a case report. AB - OBJECTIVE: to illustrate the danger of using acrylic denture as removable partial denture in a developing country. METHOD: post mortem examination was performed on a man who died after ingesting removable partial denture. RESULTS: A removable partial denture was seen seated in the mid portion of the oesophagus with its lateral wings deeply embedded in the wall of the oesophagus causing laceration and severe haemorrhage. CONCLUSION: The radiolucent nature of acrylic denture may prevent early diagnosis and intervention when it is ingested or aspirated. There is need for proper education of patients wearing partial denture in especially the acrylic type. PMID- 15827603 TI - Bacterial chromosome segregation. AB - In most bacteria two vital processes of the cell cycle: DNA replication and chromosome segregation overlap temporally. The action of replication machinery in a fixed location in the cell leads to the duplication of oriC regions, their rapid separation to the opposite halves of the cell and the duplicated chromosomes gradually moving to the same locations prior to cell division. Numerous proteins are implicated in co-replicational DNA segregation and they will be characterized in this review. The proteins SeqA, SMC/MukB, MinCDE, MreB/Mbl, RacA, FtsK/SpoIIIE playing different roles in bacterial cells are also involved in chromosome segregation. The chromosomally encoded ParAB homologs of active partitioning proteins of low-copy number plasmids are also players, not always indispensable, in the segregation of bacterial chromosomes. PMID- 15827604 TI - The Obg subfamily of bacterial GTP-binding proteins: essential proteins of largely unknown functions that are evolutionarily conserved from bacteria to humans. AB - Members of the Obg subfamily of small GTP-binding proteins (called Obg, CgtA, ObgE or YhbZ in different bacterial species) have been found in various prokaryotic and eukaryotic organisms, ranging from bacteria to humans. Although serious changes in phenotypes are observed in mutant bacteria devoid of Obg or its homologues, specific roles of these GTP-binding proteins remain largely unknown. Recent genetic and biochemical studies, as well as determination of the structures of Obg proteins from Bacillus subtilis and Thermus thermophilus, shed new light on the possible functions of the members of the Obg subfamily and may constitute a starting point for the elucidation of their exact biological role. PMID- 15827605 TI - Farnesyl diphosphate synthase; regulation of product specificity. AB - Farnesyl diphosphate synthase (FPPS) is a key enzyme in isoprenoid biosynthesis which supplies sesquiterpene precursors for several classes of essential metabolites including sterols, dolichols, ubiquinones and carotenoids as well as substrates for farnesylation and geranylgeranylation of proteins. It catalyzes the sequential head-to-tail condensation of two molecules of isopentenyl diphosphate with dimethylallyl diphosphate. The enzyme is a homodimer of subunits, typically having two aspartate-rich motifs with two sets of substrate binding sites for an allylic diphosphate and isopentenyl diphosphate per homodimer. The synthase amino-acid residues at the 4th and 5th positions before the first aspartate rich motif mainly determine product specificity. Hypothetically, type I (eukaryotic) and type II (eubacterial) FPPSs evolved from archeal geranylgeranyl diphosphate synthase by substitutions in the chain length determination region. FPPS belongs to enzymes encoded by gene families. In plants this offers the possibility of differential regulation in response to environmental changes or to herbivore or pathogen attack. PMID- 15827606 TI - Existing and future therapeutic options for hepatitis C virus infection. AB - Hepatitis C virus (HCV) infection is an important cause of chronic hepatitis, cirrhosis, hepatocellular carcinoma and liver failure worldwide. Chronic hepatitis C virus infection is treated with interferon-a (IFN-alpha), pegylated interferon-alpha (PEG-IFNalpha) alone or in combination with ribavirin; however, a significant fraction of patients either fail to respond or relapse after cessation of therapy. Efforts to identify and develop highly specific and potent HCV inhibitors have intensified recently. Each of the virally encoded replication enzymes has been a focus of studies as well as viral receptors and the host immune system. This review summarizes recent progress in the search for novel anti-HCV agents. PMID- 15827607 TI - FEDMA--a simple algorithm for theoretical modeling of linear metabolic pathways: from fuzzy data sets to prediction and experiment. AB - A theoretical model of a chain of irreversible Michaelis-Menten reactions proceeding inside a living cell, taking cell growth, division and subcellular compartmentation into account, was proposed. It became a basis for the construction of a "fuzzy" enzymatic data-modeling algorithm (FEDMA) - a procedure allowing the estimation of missing parameter values for the modeled system, in accordance both with the derived theoretical rules and the available experimental data. The obtained tool was tested to model the heme biosynthesis pathway in Saccharomyces cerevisiae, where about 40% of parameters remain unknown. The missing parameters estimated by means of FEDMA fall in the range of expected values. PMID- 15827608 TI - Co-inoculation with two non-infectious cDNA copies of potato spindle tuber viroid (PSTVd) leads to the appearance of novel fully infectious variants. AB - Potato spindle tuber viroid (PSTVd) is one of the smallest (about 360 nt) infectious plant agents. It is composed of a single-stranded circular non-coding RNA molecule. In the course of previous passage experiments with two intermediate PSTVd variants I2 and I4, three non-infectious clones (I2-50, I4-37 and I4 VI-17) were found. When inoculated separately as cDNAs on tomato "Rutgers" test plants these variants did not induce any visible disease symptoms and did not produce progeny. The presence of such non-infectious variants raises several questions about their origin and biology and to answer them, mixed co-infections with cDNA copies of two non-infectious variants (I2-50, I4-37) were performed. PSTVd infection was observed in seven out of 30 inoculated plants. The progeny isolated from three separate plants contained novel variants, together with the parental I2 and I4 sequences. It is conceivable that the appearance of repaired PSTVd molecules, clearly capable of cell-to-cell movement leading to the systemic infection, results from recombination events. An analysis of the recombinant molecules and comparison with databases identified the specific sites responsible for the restricted infectivity of the I2-50 and I4-37 PSTVd variants. In parallel experiments in which (+) strand PSTVd infectious transcripts were used, no recombinants were observed, and the original I2-50 and I4-37 non-infectious sequences were not detected in the progeny. PMID- 15827609 TI - Characterization of Bacillus subtilis clones surviving overproduction of Zeta, a pSM19035 plasmid-encoded toxin. AB - The postsegregational killing system of pSM19035 plasmid consists of the proteins Zeta and Epsilon, a toxin and an antidote, respectively. Zeta mutants were isolated with the use of Bacillus subtilis strain with the zeta gene under control of an inducible promoter integrated into the chromosome. Results of mutant analysis point to the amino terminal part of the Zeta protein as being responsible for the toxicity. PMID- 15827610 TI - Overexpression of genes involved in phytochelatin biosynthesis in Escherichia coli: effects on growth, cadmium accumulation and thiol level. AB - In Escherichia coli, heterologous production of Schizosaccharomyces pombe phytochelatin synthase (PCS) along with overproduction of E. coli serine acetyltransferase (SAT) and gamma-glutamylcysteine synthase (gammaECS) was achieved and resulted in the accumulation of phytochelatins in bacterial cells. Overproduction of either gammaECS alone or simultaneous production of all three proteins in bacterial cells were accompanied by reduced growth rate in liquid cultures. Interestingly, bacteria overproducing either gammaECS or both SAT and gammaECS (with elevated level of gamma-glutamylcysteine but not of phytochelatins) were able to accumulate more cadmium per dry weight than the control. However, the most efficient cadmium accumulation was observed in bacteria with elevated levels of all three proteins: SAT, gammaECS and PCS. Therefore, "pushing" the entire pathway might be the most promising approach in modification of bacteria for potential bioremediation purposes because the level of intermediates, cysteine and glutathione, can limit the rate of production of phytochelatins. However, in such bacteria other metabolic process might become limiting for efficient growth. PMID- 15827611 TI - Isolation of Nicotiana plumbaginifolia cDNAs encoding isoforms of serine acetyltransferase and O-acetylserine (thiol) lyase in a yeast two-hybrid system with Escherichia coli cysE and cysK genes as baits. AB - We applied the yeast two-hybrid system for screening of a cDNA library of Nicotiana plumbaginifolia for clones encoding plant proteins interacting with two proteins of Escherichia coli: serine acetyltransferase (SAT, the product of cysE gene) and O-acetylserine (thiol)lyase A, also termed cysteine synthase (OASTL-A, the product of cysK gene). Two plant cDNA clones were identified when using the cysE gene as a bait. These clones encode a probable cytosolic isoform of OASTL and an organellar isoform of SAT, respectively, as indicated by evolutionary trees. The second clone, encoding SAT, was identified independently also as a "prey" when using cysK as a bait. Our results reveal the possibility of applying the two-hybrid system for cloning of plant cDNAs encoding enzymes of the cysteine synthase complex in the two-hybrid system. Additionally, using genome walking sequences located upstream of the sat1 cDNA were identified. Subsequently, in silico analyses were performed aiming towards identification of the potential signal peptide and possible location of the deduced mature protein encoded by sat1. PMID- 15827612 TI - Translational readthrough of a termination codon in the yeast mitochondrial mRNA VAR1 as a result of mutation in the release factor mRF1. AB - Yeast mitochondrial DNA codes for eight major polypeptides. Translation of he mitochondrially encoded polypeptides in strains with mutated mitochondrial release factor, mRF1, was found to result in the synthesis of a novel protein, V2. Different mrf1 alleles were associated with different efficiency of V2p synthesis. Translation of V2p was enhanced by paromomycin. Comparative analysis of peptides resulting from protease digestion indicated that V2p is a derivative of Var1p. According to our hypothesis, V2p represents a readthrough product of the natural stop codon in VAR1 mRNA. PMID- 15827613 TI - Effect of SOS-induced Pol II, Pol IV, and Pol V DNA polymerases on UV-induced mutagenesis and MFD repair in Escherichia coli cells. AB - Irradiation of organisms with UV light produces genotoxic and mutagenic lesions in DNA. Replication through these lesions (translesion DNA synthesis, TSL) in Escherichia coli requires polymerase V (Pol V) and polymerase III (Pol III) holoenzyme. However, some evidence indicates that in the absence of Pol V, and with Pol III inactivated in its proofreading activity by the mutD5 mutation, efficient TSL takes place. The aim of this work was to estimate the involvement of SOS-inducible DNA polymerases, Pol II, Pol IV and Pol V, in UV mutagenesis and in mutation frequency decline (MFD), a mechanism of repair of UV-induced damage to DNA under conditions of arrested protein synthesis. Using the argE3-->Arg(+) reversion to prototrophy system in E. coli AB1157, we found that the umuDC encoded Pol V is the only SOS-inducible polymerase required for UV mutagenesis, since in its absence the level of Arg(+) revertants is extremely low and independent of Pol II and/or Pol IV. The low level of UV-induced Arg(+) revertants observed in the AB1157mutD5DumuDC strain indicates that under conditions of disturbed proofreading activity of Pol III and lack of Pol V, UV induced lesions are bypassed without inducing mutations. The presented results also indicate that Pol V may provide substrates for MFD repair; moreover, we suggest that only those DNA lesions which result from umuDC-directed UV mutagenesis are subject to MFD repair. PMID- 15827614 TI - Mismatch dependent uracil/thymine-DNA glycosylases excise exocyclic hydroxyethano and hydroxypropano cytosine adducts. AB - Exocyclic adducts of DNA bases, such as etheno- and hydroxyalkano- ones, are generated by a variety of bifunctional agents, including endogenously formed products of lipid peroxidation. In this work we selectively modified cytosines in the 5'-d(TTT TTT CTT TTT CTT TTT CTT TTT T)-3' oligonucleotide using: chloroacetaldehyde to obtain 3,N(4)-alpha-hydroxyethano- (HEC) and 3,N(4)-etheno- (epsilonC), acrolein to obtain 3,N(4)-alpha-hydroxypropano- (HPC) and crotonaldehyde to obtain 3,N(4)-alpha-hydroxy-gamma-methylpropano- (mHPC) adducts of cytosine. The studied adducts are alkali-labile which results in oligonucleotide strain breaks at the sites of modification upon strong base treatment. The oligonucleotides carrying adducted cytosines were studied as substrates of Escherichia coli Mug, human TDG and fission yeast Thp1p glycosylases. All the adducts studied are excised by bacterial Mug although with various efficiency: epsilonC >HEC >HPC >mHPC. The yeast enzyme excises efficiently epsilonC>HEC>HPC, whereas the human enzyme excises only epsilonC. The pH-dependence curves of excision of eC, HEC and HPC by Mug are bell shaped and the most efficient excision of adducts occurs within the pH range of 8.6-9.6. The observed increase of excision of HEC and HPC above pH 7.2 can be explained by deprotonation of these adducts, which are high pK(a) compounds and exist in a protonated form at neutrality. On the other hand, since epsilonC is in a neutral form in the pH range studied, we postulate an involvement of an additional catalytic factor. We hypothesize that the enzyme structure undergoes a pH-induced rearrangement allowing the participation of Lys68 of Mug in catalysis via a hydrogen bond interaction of its epsilon-amino group with N(4) of the cytosine exocyclic adducts. PMID- 15827615 TI - Inhibition of DNA repair glycosylases by base analogs and tryptophan pyrolysate, Trp-P-1. AB - DNA base analogs, 2,4,5,6-substituted pyrimidines and 2,6-substituted purines were tested as potential inhibitors of E. coli Fpg protein (formamidopyrimidine DNA glycosylase). Three of the seventeen compounds tested revealed inhibitory properties. 2-Thioxanthine was the most efficient, inhibiting 50% of 2,6-diamino 4-hydroxy-5N-methyl-formamidopyrimidine (Fapy-7MeG) excision activity at 17.1 microM concentration. The measured K(i) was 4.44 +/- 0.15 microM. Inhibition was observed only when the Fpg protein was first challenged to its substrate followed by the addition of the base analog, suggesting uncompetitive (catalytic) inhibition. For two other compounds, 2-thio- or 2-oxo-4,5,6-substituted pyrimidines, IC(50) was only 343.3 +/- 58.6 and 350 +/- 24.4 microM, respectively. No change of the Fpg glycosylase activity was detected in the presence of Fapy-7MeG, up to 5 microM. We also investigated the effect of DNA structure modified by tryptophan pyrolysate (Trp-P-1) on the activity of base excision repair enzymes: Escherichia coli and human DNA glycosylases of oxidized (Fpg, Nth) and alkylated bases (TagA, AlkA, and ANPG), and for bacterial AP endonuclease (Xth protein). Trp-P-1, which changes the secondary DNA structure into non-B, non-Z most efficiently inhibited excision of alkylated bases by the AlkA glycosylase (IC(50) = 1 microM). The ANPG, TagA, and Fpg proteins were also inhibited although to a lesser extent (IC(50) = 76.5 microM, 96 microM, and 187.5 microM, respectively). Trp-P-1 also inhibited incision of DNA at abasic sites by the beta-lyase activity of the Fpg and Nth proteins, and to a lesser extent by the Xth AP endonuclease. Thus, DNA conformation is critical for excision of damaged bases and incision of abasic sites by DNA repair enzymes. PMID- 15827616 TI - Apoptosis induced by membrane damage in human lymphocytes; effects of arachidonic acid and its photoproducts. AB - The effect of arachidonic acid (AA) combined with UVA irradiation was studied in a model system mimicking phototherapy PUVA (psoralen+UVA) ex vivo in vitro. The contribution of damage to the plasma membrane by PUVA was tested on human lymphocytes derived from healthy donors. The effect of arachidonic acid (AA) combined with UVA irradiation was compared with that of a psoralen photoadduct to AA added to the culture. The adduct, obtained photochemically and purified, was characterized by NMR and MS spectrometry as a cycloadduct of psoralen to the vinylene bond of the acid (AA<>PSO). The reactions of cultured cells, manifested 20 h after treatment by changes in apoptosis and mitochondrial depolarization, were monitored by flow cytometry by tagging lymphocytes with appropriate fluorescent probes. Treatment of lymphocyte suspension within AA doses from 40 to 100 microM gradually induced a shift from Anx-V(+) (single positive cells) to late apoptotic, Anx-V(+)PI(+) (double positive cells) in a dose dependent manner. The adduct, AAPSO, induced apoptotic changes at a concentration 2-3 times higher than free AA. Combination of psoralen (1 microM ) or arachidonic acid (20-120 microM) with UVA irradiation (2-6 J/cm(2)) accelerated the plasma membrane changes in a synergic way. Preliminary studies indicated that changes in the transmembrane potential of mitochondria paralleled the apoptosis when cells were treated by AA alone. Our findings showed that UVA radiation of lymphocytes in the presence of arachidonic acid, as in the presence of psoralen, enhanced apoptosis of cells in a synergic manner. Thus, PUVA-induced apoptosis may proceed in part by a still undefined signaling pathway(s) triggered in lymphocyte membranes. PMID- 15827617 TI - Protein production and secretion in an Aspergillus nidulans mutant impaired in glycosylation. AB - O-glycosylation has been considered a limiting factor in protein secretion in filamentous fungi. Overexpression of the yeast DPM1 gene encoding dolichylphosphate mannose synthase (DPMS) in an Aspergillus nidulans mutant (BWB26A) deficient in O-glycosylation caused an increase in the number of secretory vesicles and changes in protein secretion. However, the secretory proteins, primarily O-mannosylated glucoamylase and N-glycosylated invertase, were mainly trapped in the periplasmic space. Different glycoforms of invertase were found insite the cells, in the periplasmic space and in the cultivation medium. Our data point to the importance of the cell wall as a barrier in protein secretion. PMID- 15827618 TI - Rsp5 ubiquitin ligase affects isoprenoid pathway and cell wall organization in S. cerevisiae. AB - Dimethylallyl diphosphate, an isomer of isopentenyl diphosphate, is a common substrate of Mod5p, a tRNA modifying enzyme, and the farnesyl diphosphate synthase Erg20p, the key enzyme of the isoprenoid pathway. rsp5 mutants, defective in the Rsp5 ubiquitin-protein ligase, were isolated and characterized as altering the mitochondrial/cytosolic distribution of Mod5p. To understand better how competition for the substrate determines the regulation at the molecular level, we analyzed the effect of the rsp5-13 mutation on Erg20p expression. The level of Erg20p was three times lower in rsp5-13 compared to the wild type strain and this effect was dependent on active Mod5p. Northern blot analysis indicated a regulatory role of Rsp5p in ERG20 transcription. ERG20 expression was also impaired in pkc1Delta lacking a component of the cell wall integrity signaling pathway. Low expression of Erg20p in rsp5 cells was accompanied by low level of ergosterol, the main end product of the isoprenoid pathway. Additionally, rsp5 strains were resistant to nystatin, which binds to ergosterol present in the plasma membrane, and sensitive to calcofluor white, a drug destabilizing cell wall integrity by binding to chitin. Furthermore, the cell wall structure appeared abnormal in most rsp5-13 cells investigated by electron microscopy and chitin level in the cell wall was increased two-fold. These results indicate that Rsp5p affects the isoprenoid pathway which has important roles in ergosterol biosynthesis, protein glycosylation and transport and in this way may influence the composition of the plasma membrane and cell wall. PMID- 15827619 TI - Functional relationships between the Saccharomyces cerevisiae cis prenyltransferases required for dolichol biosynthesis. AB - In the yeast Saccharomyces cerevisiae the RER2 and SRT1 genes encode Rer2 and Srt1 proteins with cis-prenyltransferase (cis-PT-ase) activity. Both cis-PT-ases utilize farnesyl diphosphate (FPP) as a starter for polyprenyl diphosphate (dolichol backbone) formation. The products of the Rer2 and Srt1 proteins consist of 14-17 and 18-23 isoprene units, respectively. In this work we demonstrate that deletion or overexpression of SRT1 up-regulates the activity of Rer2p and dolichol content. However, upon overexpression of SRT1, preferential synthesis of longer-chain dolichols and a decrease in the amount of the shorter species are observed. Furthermore, overexpression of the ERG20 gene (encoding farnesyl diphosphate synthase, Erg20p) induces transcription of SRT1 mRNA and increases the levels of mRNA for RER2 and DPM1 (dolichyl phosphate mannose synthase, Dpm1p). Subsequently the enzymatic activity of Rer2p and dolichol content are also increased. However, the amount of Dpm1p or its enzymatic activity remain unchanged. PMID- 15827620 TI - Light conditions alter accumulation of long chain polyprenols in leaves of trees and shrubs throughout the vegetation season. AB - In many plants belonging to angiosperms and gymnosperms the accumulation in leaves of long chain polyprenols and polyprenyl esters during growth in natural habitats depends on the light intensity. The amount of polyprenols in leaves is also positively correlated with the thickness of the leaf blade (SLA, specific leaf area). The polyprenol content of leaves shows seasonal changes with a maximum in autumn and a minimum in early summer with the difference between poorly and well illuminated plants persisting throughout the vegetation season. PMID- 15827621 TI - The occurrence of polyprenols in seeds and leaves of woody plants. AB - The contents of the heterogenous group of polyisoprenoids was found about two orders of magnitude lower in seeds than the amount of polyprenols and/or their carboxylic esters accumulated during vegetation season in leaves. In contrast to leaves, no seeds were found containing more than 0.5 mg of these lipids per gram of dry tissue. Almost 50% had less than 0.01 mg/g - the amount which is the limit of detection by the procedure used in this work. In gymnosperms (10 representatives of Cupressaceae, Pinaceae and Taxaceae) the polyprenol spectra in seeds and in needles were similar. In angiosperms (25 representatives of 13 botanical families) the polyisoprenoid mixture in seeds resembled the minor, additional subfamily found in leaves. PMID- 15827622 TI - Dolichols of the fern Matteucia struthiopteris. AB - Dolichols isolated from leaves of the fern Matteucia struthiopteris were present as a mixture of prenologues composed of 14 up to 20 isoprene units with Dol-16 dominating. They comprised approximately 0.004% of the fresh weight of fresh plant tissue and were accompanied by traces of polyprenols (Pren-14 up to Pren 17, Pren-16 dominating). Their structure was confirmed by electropray ionization mass spectrometry (ESI-MS). This is the first time that dolichols have been reported as dominating polyisoprenoid alcohols in plant photosynthetic tissue. PMID- 15827623 TI - Genetic immunization of ducks for production of antibodies specific to Helicobacter pylori UreB in egg yolks. AB - Following genetic immunization of laying ducks with a plasmid expressing Helicobacter pylori UreB (large subunit of urease), IgY against UreB were obtained from egg yolks. These polyclonal and monospecific IgY antibodies are of higher-titer and specifically recognize recombinant H. pylori urease purified from Escherichia coli. To our knowledge this is the first report describing generation of IgY antibodies directed against antigens of H. pylori by DNA-based immunization. PMID- 15827624 TI - MOFOID--not only the protein modeling server. AB - MOFOID is a new server developed mainly for automated modeling of protein structures by their homology to the structures deposited in the PDB database. Selection of a template and calculation of the alignment is performed with the Smith-Waterman or Needleman-Wunsch algorithms implemented in the EMBOSS package. The final model is built and optimised with programs from the JACKAL package. The wide spectrum of options in the web-based interface and the possibility of uploading user's own alignment make MOFOID a suitable platform for testing new approaches in the alignment building. The server is available at https:// valis.ibb.waw.pl/mofoid/. PMID- 15827625 TI - [3 + 3] Cycloadditions and related strategies in alkaloid natural product synthesis. AB - In this article we describe a strategically unusual approach to piperidines via formal [3 + 3] cycloaddition reactions. The scope and limitations of these processes are outlined and their employment in the synthesis of alkaloid natural products is delineated. PMID- 15827626 TI - Functionalized foldamers: synthesis and characterization of a glycosylated beta peptide 314-helix conveying the TN-antigen. AB - Herein we describe the design, synthesis, and solution structure of a novel type of conjugate composed of a naturally occurring bio-active ligand bound to an artificial peptidomimetic backbone; in this first report on such functionalized foldamers we utilized a beta-peptide as backbone and a GalNAc carbohydrate residue as ligand. PMID- 15827627 TI - A chiral molecular recognition approach to the formation of optically active quaternary centres in aza-Henry reactions. AB - An approach to asymmetric catalysis based on chiral molecular recognition by the combination of chiral Lewis acids and chiral organocatalysis for the formation of optically active quarternary centers in the aza-Henry reaction is presented; this procedure leads to products with up to 98% ee and a diastereomeric ratio of 14 : 1 in excellent yields with catalyst loadings of 5 mol%. PMID- 15827628 TI - Ethyl (benzothiazol-2-ylsulfonyl)acetate: a new reagent for the stereoselective synthesis of alpha,beta-unsaturated esters from aldehydes. AB - The title reagent engaged in the modified Julia olefination with aldehydes under mild reaction conditions (DBU, CH(2)Cl(2), rt or -78 degrees C) to yield alpha,beta-unsaturated esters; aryl aldehydes and aliphatic aldehydes possessing significant chain branching elements gave trans alkene products with high stereoselectivity (E : Z up to >98 : 2), while straight chain aliphatic aldehydes gave cis products preferentially (Z : E up to 92 : 8). PMID- 15827629 TI - Selective five- and six-membered cyclic amine syntheses via capture of episulfonium ions. AB - Amide nitrogens open episulfonium ions to form pyrrolidines or piperidines selectively, depending on the nitrogen substituent, in either reversible or irreversible reactions. PMID- 15827630 TI - Total synthesis of paracentrone, C31-allenic apo-carotenoid. AB - The stereocontrolled total synthesis of a C31-allenic apo-carotenoid, paracentrone, was achieved by the convergent C20 + C11 = C31 strategy. The key elements of our synthesis were the Pd-catalyzed cross-coupling to stereoselectively construct the conjugated polyene backbone skeleton and the designed geometrical isomerization at the central double bond of the conjugated polyene chain. In addition, the terminal oxygenated cyclohexane ring having the allenic moiety was prepared by the highly diastereoselective Sharpless epoxidation under our own reaction conditions. PMID- 15827631 TI - Investigation of the aqueous transmetalation of pi-allylpalladium with indium salt: the use of the Pd(OAc)2-TPPTS catalyst. AB - pi-Allylpalladium complexes could be generated in water by the palladium(0) water soluble catalyst prepared in situ from palladium acetate and TPPTS. These complexes were transmetalated with indium to react with benzaldehyde. The aqueous solution of Pd(0)(TPPTS)(n) could be reused without deterioration of the catalyst in the first and second recycling. The system proved to be efficient with primary and secondary allylic substrates. The stereochemical outcome of the allylation through umpolung of allylpalladium, was also studied using models with a restraint conformation. PMID- 15827632 TI - The relative orientation of the lipid and carbohydrate moieties of lipochitooligosaccharides related to nodulation factors depends on lipid chain saturation. AB - Lipochitooligosaccharides (LCOs) signal the symbiosis of rhizobia with legumes and the formation of nitrogen-fixing root nodules. LCOs 1 and 2 share identical tetrasaccharide scaffolds but different lipid moieties (1, LCO-IV(C16:1[9Z], SNa) and , LCO-IV(C16:2[2E,9Z], SNa)). The conformational behaviors of both LCOs were studied by molecular modeling and NMR. Modeling predicts that a small lipid modification would result in a different relative orientation of the lipid and tetrasaccharide moieties. Diffusion ordered spectroscopy reports that both LCOs form small aggregates above 1 mM. Nuclear Overhauser spectroscopy (NOESY) data, collected under monomeric conditions, reveals lipid-carbohydrate contacts only for 1, in agreement with the modeling data. The distinct molecular structures of 1 and 2 have the potential to contribute to their selective binding by legume proteins. PMID- 15827633 TI - Boron dipyrromethene fluorophore based fluorescence sensor for the selective imaging of Zn(II) in living cells. AB - A simple PET fluorescence sensor (BDA) for Zn2+ that utilizes 1,3,5,7-tetramethyl boron dipyrromethene as a reporting group and di(2-picolyl)amine as a chelator for Zn2+ has been synthesized and characterized. BDA has an excitation (491 nm) and emission wavelength (509 nm) in the visible range. The fluorescence quantum yields of the zinc-free and zinc-bound states of BDA are 0.077 and 0.857, respectively. With a low pKa of 2.1 +/- 0.1, BDA has the advantage of less sensitivity to pH than fluorescein-based Zn2+ sensors, and the fluorescence emission of zinc-binding is pH-independent in the range of pH 3-10. Under physiological conditions, metal ions such as Na+, K+, Ca2+, Mg2+, Mn2+ and Fe2+ have little interference. The apparent dissociation constant (Kd) is 1.0 +/- 0.1 nM. Using fluorescence microscopy, the sensor is shown to be capable of imaging intracellular Zn2+ changes. PMID- 15827634 TI - Branched [n]rotaxanes (n = 2-4) from multiple dibenzo-24-crown-8 ether wheels and 1,2-bis(4,4'-dipyridinium)ethane axles. AB - To investigate the possibility of incorporating the 1,2 bis(pyridinium)ethane[subset or is implied by]24C8 [2]pseudorotaxane motif into dendrimer like macromolecules, a series of branched [n]rotaxanes were prepared employing multiple dibenzo-24-membered crown ether wheels with various aromatic core structures and the 1,2-bis(4,4'-dipyridinium)ethane axle. Yields of branched [2]-, [3]- and [4]rotaxanes were dependent on the size of the core and the relative proximity of the crown ethers arranged around the core unit. PMID- 15827635 TI - 4-(Tetrahydro-4H-thiopyran-1-oxide-4-ylidene)-cyclohexanone oxime in the solid state. A two-dimensional network of enantiomorphous chains interconnected by weak hydrogen bonds. AB - From a saturated C6H6 solution of racemic 4-(tetrahydro-4H-thiopyran-1-oxide-4 ylidene)-cyclohexanone oxime [1(1-R/1-S)] the co-crystal is crystallized. Single crystal X-ray analysis showed that (1)4.C6H6 (P1 space group) in the solid-state consists of enantiomorphous, non-covalent polymer-like chains that contain, in an alternating fashion, the crystallographically independent enantiomers 1-R and 1 R' or 1-S and 1-S'', respectively. Within each chain the enantiomers are linked by 'head-to-tail' intermolecular oxime-sulfoxide hydrogen bonding [D(2) motif]. Neighbouring chains consist of enantiomers with opposite configuration and possess opposite molecular 'head-to-tail' alignments. The enantiomorphous chains are interconnected by weak intermolecular C-HO hydrogen bonds involving centrosymmetric C-Hoxime [R(12)] and C-Hsulfoxide [R(8)] motifs between the 1-R and 1-S molecules in neighbouring chains; a nearly planar two-dimensional hydrogen bonding network motif is obtained. In the crystallographic direction [1 0 0] the layers stack in such a fashion that chains occupying successive layers with an identical 'head-to-tail' alignment are positioned on top of each other. Concomitantly, channels with areas of ca. 25 Angstroms(2) are obtained, which are occupied by C6H6 solvent molecules. A comparison of the IR and Raman spectra of with those obtained for native 1 that does not contain C6H6, indicates that intermolecular oxime-sulfoxide hydrogen bonding [D(2) motif] also occurs for native 1 in the solid-state. PMID- 15827636 TI - The neomycin biosynthetic gene cluster of Streptomyces fradiae NCIMB 8233: characterisation of an aminotransferase involved in the formation of 2 deoxystreptamine. AB - The biosynthetic gene cluster of the 2-deoxystreptamine (DOS)-containing aminoglycoside antibiotic neomycin has been cloned for the first time by screening of a cosmid library of Streptomyces fradiae NCIMB 8233. Sequence analysis has identified 21 putative open reading frames (ORFs) in the neomycin gene cluster (neo) with significant protein sequence similarity to gene products involved in the biosynthesis of other DOS-containing aminoglycosides, namely butirosin (btr), gentamycin (gnt), tobramycin (tbm) and kanamycin (kan). Located at the 5'-end of the neo gene cluster is the previously-characterised neomycin phosphotransferase gene (apH). Three genes unique to the neo and btr clusters have been revealed by comparison of the neo cluster to btr, gnt, tbm and kan clusters. This suggests that these three genes may be involved in the transfer of a ribose moiety to the DOS ring during the antibiotic biosynthesis. The product of the neo-6 gene is characterised here as the L-glutamine : 2-deoxy-scyllo inosose aminotransferase responsible for the first transamination in DOS biosynthesis, which supports the assignment of the gene cluster. PMID- 15827637 TI - Carboxylate-stabilised sulfur ylides (thetin salts) in asymmetric epoxidation for the synthesis of glycidic acids. Mechanism and implications. AB - The reaction of carboxylate-stabilised sulfur ylides (thetin salts) with aldehydes and ketones has been investigated. Using both achiral and chiral sulfur ylides, good yields were obtained with dimsylsodium or LHMDS as bases in DMSO or THF-DMSO mixtures. However, the enantioselectivities observed with a camphor based sulfide were only moderate (up to 67%). The reaction was studied mechanistically by independent generation of the betaine (via the hydroxyl sulfonium salt) in the presence of a more reactive aldehyde, which resulted in incorporation of the more reactive aldehyde and showed that betaine formation was reversible. Thus, the moderate enantiomeric excess observed is a consequence of the enantiodifferentiating step being the ring closure step rather than the betaine forming step. We had expected betaine formation might be non-reversible because a carboxylate-stabilised ylide has only slightly higher stability than a phenyl-stabilised ylide, which does largely react non-reversibly with aldehydes. Evidently, a carboxylate-stabilised ylide is significantly more stable than a phenyl-stabilised ylide and as such reacts reversibly with aldehydes. PMID- 15827638 TI - Calix[4]azacrown and 4-aminophthalimide-appended calix[4]azacrown: synthesis, structure, complexation and fluorescence signaling behaviour. AB - The synthesis and single crystal structure of 25,27-(diamidomonoazacrown-5) calix[4]arene (L), calix[4]arene capped by a diamide bridge in the 1,3-position on the lower rim, which forms a supramolecular dimer in the solid state via intermolecular hydrogen bonding, are described. A 4-aminophthalimide (AP) fluorophore has been regioselectively linked to the secondary amino function of the azacrown unit with a dimethylene spacer to construct N-(4 aminophthalimidoethyl)calix[4]azacrown (APL), a fluorescent sensor, via a fluorophore-spacer-receptor architecture. Fluorescence quantum yield and lifetime of APL have been measured to be lower than those of the bare fluorophore (AP) due to the photoinduced intramolecular electron transfer (PIET) between the fluorophore and the receptor moieties of the molecule. In the presence of transition metal ions, fluorescence enhancement of is observed suggesting the binding of the metal ion to the sensor. Complexation properties of APL with transition metal ions are investigated using UV-vis spectroscopy. A 1 : 1 stoichiometry of the complex is determined from a Job plot and the corresponding association constants for the various metal ions are evaluated. Fe(3+) and Cu(2+) ions have the largest association constants (K= 2.3 x 10(5) M(-1) and 1.6 x 10(5) M(-1) respectively) compared to other metal ions indicating that they form complexes selectively with APL. PMID- 15827639 TI - Asymmetric conjugate reductions with samarium diiodide: asymmetric synthesis of (2S,3R)- and (2S,3S)-[2-2H,3-2H]-leucine-(S)-phenylalanine dipeptides and (2S,3R) [2-(2)H,3-2H]-phenylalanine methyl ester. AB - The highly diastereoselective samarium diiodide and D(2)O-promoted conjugate reduction of homochiral (E)- and (Z)-benzylidene and isobutylidene diketopiperazines (E)-5,7 and (Z)-6,8 has been demonstrated. This methodology allows the asymmetric synthesis of methyl (2S,3R)-dideuteriophenylalanine 27 in > or = 95% de and >98% ee, and (2S,3R)- or (2S,3S)-dideuterioleucine-(S) phenylalanine dipeptides 37 and 38 in moderate de, 66% and 74% respectively. A mechanism is proposed to account for this process. PMID- 15827640 TI - Orientational isomers of alpha-cyclodextrin [2]semi-rotaxanes with asymmetric dicationic threads. AB - Two series of novel dicationic threading molecules [Quin(CH2)10R]2+ and [3,5 Lut(CH2)10R]2+, where Quin+ = quinuclidinium, 3,5-Lut+ = 3,5-lutidinium, and R+ = N(CH3)3+ and N(CH3)2CH2CH3+, form [2]semi-rotaxanes with [small alpha] cyclodextrin (alpha-CD) in aqueous solution. The quinuclidinium and 3,5 lutidinium are sufficiently bulky to prevent threading while the R+ groups allow for slow threading by alpha-CD at 25 degrees C. The resulting [2]semi-rotaxanes exist in two orientational isomers owing to the asymmetry of both the alpha-CD cavity and the threading molecules. Two-dimensional 1H NMR spectroscopy and kinetics experiments reveal that the isomer in which the narrower rim (primary OHs) is positioned near the R+ group is the kinetically preferred isomer, while the other isomer is the thermodynamically preferred product. The kinetics and mechanism of the formation, dissociation, and interconversion of the two isomers have been determined at 25 degrees C. PMID- 15827641 TI - Contribution of the intramolecular hydrogen bond to the shift of the pKa value and the oxidation potential of phenols and phenolate anions. AB - Intramolecularly OHO[double bond, length as m-dash]C hydrogen bonded phenols, 2 HO-C6H2-3,5-(t-Bu)2-CONH-t-Bu (1-OH), 2-HO-C6H2-5-t-Bu-1,3-(CONH-t-Bu)2 (2-OH) and 2-HO-C6H2-3,5-(t-Bu)2-NHCO-t-Bu (4-OH), were synthesized and their phenolate anions were prepared as tetraethylammonium salts (-1O-(NEt4+), 2-O-(NEt4+) and 4 O-(NEt4+)) with intramolecular NHO(oxyanion) hydrogen bonds. 4-HO-C(6)H(2)-3,5-t Bu(2)-CONH-t-Bu (3-OH) and its phenolate anion, 3-O-(NEt4+), were synthesized as non-hydrogen bonded references. The presence of intramolecular hydrogen bonds was established through the crystallographic analysis and/or (1)H NMR spectroscopic results. Intramolecular NHO(phenol) hydrogen bonds shift the pK(a) of the phenol to a more acidic value. The results of cyclic voltammetry show that the intramolecular OH...O=C hydrogen bond negatively shifts the oxidation potential of the phenol. In contrast, the intramolecular NHO(oxyanion) hydrogen bond positively shifts the oxidation potential of the phenolate anion, preventing oxidation. These contributions of the hydrogen bond to the pKa value and the oxidation potentials probably play an important role in the formation of a tyrosyl radical in photosystem II. PMID- 15827642 TI - Synthesis of heteroarenes using cascade radical cyclisation via iminyl radicals. AB - Cascade radical cyclisation involving homolytic aromatic substitution has been used to synthesise new tetracycles. Treatment of vinyl iodide radical precursors with Me(3)Sn. radicals (from hexamethylditin) yielded intermediate vinyl radicals which undergo 5-exo cyclisation onto suitably placed nitrile groups to yield intermediate iminyl radicals. The iminyl radicals undergo aromatic homolytic substitution via 6-endo cyclisation (or 5-exo cyclisation followed by neophyl rearrangement) with loss of hydrogen (H.) in a H-abstraction step. We propose that this abstraction was facilitated by tert-butoxyl (t-BuO.) radicals from di tert-butyl peroxide or methyl radicals, generated from breakdown of trimethylstannyl radicals (Me(3)Sn.). The biologically active alkaloids mappicine and luotonin A were synthesised using the new methodology. A novel radical conversion of nitriles to primary amides is proposed. PMID- 15827643 TI - Catalysis of the ethanolysis of aryl methyl phenyl phosphinate esters by alkali metal ions: transition state structures for uncatalyzed and metal ion-catalyzed reactions. AB - This paper reports on a spectrophotometric kinetic study of the effects of the alkali metal ions Li+ and K+ on the ethanolysis of the aryl methyl phenyl phosphinate esters 3a-f in anhydrous ethanol at 25 degrees C. Rate data obtained in the absence and presence of complexing agents afford the second-order rate constants for the reaction of free ethoxide (k(EtO-)) and metal ion-ethoxide ion pairs (k(MOEt)). The sequence k(EtO-) < k(MOEt) is established for all the substrates, contrary to the generally observed reactivity order in nucleophilic substitution processes. The quantities deltaG(ip), deltaG(ts) and DeltaG(cat), which quantify the observed alkali metal ion effect in terms of transition state stabilization through chelation of the metal ion, give the order deltaG(ts) > deltaG(ip) for Li+ and K+. Hammett plots show significantly better correlation of rates with sigma and sigma(o) substituent constants than with sigma-, yielding moderately large rho(rho(o)) values that are consistent with a stepwise mechanism in which formation of a pentacoordinate (phosphorane) intermediate is the rate limiting step. The range of the values of the selectivity parameter, rho(n) (= rho]/rho(eq)), 1.3-1.6, obtained for the uncatalyzed and alkali metal ion catalyzed reactions indicates that there is no significant perturbation of the transition state (TS) structure upon chelation of the metal ions. This finding is relevant to the mechanism of enzymatic phosphoryl transfer involving metal ion co factors. The present results enable one to compare structural effects for nucleophilic reactions of several series of organophosphorus substrates. It is shown that the order of reactivity of the substrates: 4-nitrophenyl dimethyl phosphinate (2) > 3a > 4-nitrophenyl diphenyl phosphinate (1) is determined mainly by the steric effects of the alkyl/aryl substituents around the central P atom in the TS of the reaction. PMID- 15827644 TI - Investigation of the interaction between peanut agglutinin and synthetic glycopolymeric multivalent ligands. AB - The interaction between synthetic glycoplymers bearing beta-D-galactose side groups and the lectin peanut agglutinin (PNA) was investigated by UV-difference spectroscopy and isothermal titration calorimetry (ITC). UV-difference spectroscopy indicated that the polymer-lectin interaction was stronger than that between PNA and either the corresponding monomer, D-galactose or D-lactose. The thermodynamics of binding (K, DeltaG, DeltaH, DeltaS and n) were determined from ITC data by fitting with a two-site, non-cooperative binding model. It was found that the glycopolymer displayed around a 50 times greater affinity for the lectin than the parent carbohydrate, and around 10 times greater than the monomer, on a valency-corrected basis. Binding was found to be entropically driven, and was accompanied by aggregation and precipitation of protein molecules. Furthermore, interesting differences between polymers prepared either from deacetylated monomers, or by deacetylation of pre-formed polymers, were found. PMID- 15827645 TI - Cyclodextrin and modified cyclodextrin complexes of E-4-tert-butylphenyl-4' oxyazobenzene: UV-visible, 1H NMR and ab initio studies. AB - alpha-Cyclodextrin, beta-cyclodextrin, N-(6(A)-deoxy-alpha-cyclodextrin-6(A)-yl) N'6(A)-deoxy-beta-cyclodextrin-6(A)-yl)urea and N,N-bis(6(A)-deoxy-beta cyclodextrin-6(A)-yl)urea (alphaCD, betaCD, 1 and 2) form inclusion complexes with E-4-tert-butylphenyl-4'-oxyazobenzene, E-3(-). In aqueous solution at pH 10.0, 298.2 K and I = 0.10 mol dm(-3)(NaClO(4)) spectrophotometric UV-visible studies yield the sequential formation constants: K(11) = (2.83 +/- 0.28) x 10(5) dm(3) mol(-1) for alphaCD.E-(-), K(21) = (6.93 +/- 0.06) x 10(3) dm(3) mol(-1) for (alphaCD)(2).E-3(-), K(11) = (1.24 +/- 0.12) x 10(5) dm(3) mol(-1) for betaCD.E-(-), K(21) = (1.22 +/- 0.06) x 10(4) dm(3) mol(-1) for (betaCD)(2).E-( ), K(11) = (3.08 +/- 0.03) x 10(5) dm(3) mol(-1) for .E-3(-), K(11) = (8.05 +/- 0.63) x 10(4) dm(3) mol(-1) for .E-3(-) and K(12) = (2.42 +/- 0.53) x 10(4) dm(3) mol(-1) for .(E-3(-))(2). (1)H ROESY NMR studies show that complexation of E-3(-) in the annuli of alphaCD, betaCD, 1 and 2 occurs. A variable-temperature (1)H NMR study yields k(298 K)= 6.7 +/- 0.5 and 5.7 +/- 0.5 s(-1), DeltaH = 61.7 +/- 2.7 and 88.1 +/- 4.2 kJ mol(-1) and DeltaS = -22.2 +/- 8.7 and 65 +/- 13 J K(-1) mol( 1) for the interconversion of the dominant includomers (complexes with different orientations of alphaCD) of alphaCD.E-3(-) and (alphaCD)(2).E-3(-), respectively. The existence of E-3(-) as the sole isomer was investigated through an ab initio study. PMID- 15827646 TI - Polycyclic compounds from aminopolyols and alpha-dicarbonyls: structure and application in the synthesis of exoditopic ligands. AB - The structure and stereochemistry of the crystalline 2 : 2 condensation product ("glytham") of glyoxal and tris(hydroxymethyl)aminomethane was conclusively determined by X-ray diffractometric analysis. The singular disposition of the heteroatoms suggests the employment of glytham as starting material for the synthesis of ditopic ligands for metal ions. Some derivatives of glytham were prepared and their binding properties towards alkaline metal ions were preliminarily investigated by ESI-MS and NMR. PMID- 15827647 TI - Urea vs. thiourea in anion recognition. AB - Neutral anion receptors (LH) form stable 1 : 1 H-bond [LH...X]- complexes with carboxylates, halides and phosphate (X-). Some of the [LH...X]- complexes, in presence of an excess of X-, release an HX fragment, with formation of [HX2]- and the deprotonated receptor L-. The tendency towards deprotonation increases with the acidity of the receptor and with the stability of the [HX2]- self-complex. Thus, the more acidic thiourea containing receptor deprotonates in the presence all the investigated anions except chloride, whereas the less acidic urea containing receptor undergoes deprotonation only in the presence of fluoride, due to the high stability of [HF2]-. PMID- 15827648 TI - Glucuronidation of steroidal alcohols using iodosugar and imidate donors. AB - We report a study of the glucuronidation of a number of important steroidal secondary alcohols. The alcohols studied are androsterone 7, epiandrosterone 8, 17-acetoxy-androstane-3alpha,17beta-diol 9, 11alpha-hydroxyprogesterone 10, and 3 benzoylestradiol 11. These were first glucuronidated using the Schmidt trichloroacetimidate method with variations in acyl substituent (viz. derivatives 2 and 3), Lewis acid catalyst and order of addition. The results are contrasted with those obtained using our recently described glycosyl iodide donor 4, catalysed either by N-iodosuccinimide (NIS) or various metal salts. PMID- 15827649 TI - On the utility of the azido transfer protocol: synthesis of 2- and 5-azido N methylimidazoles, 1,3-thiazoles and N-methylpyrazole and their conversion to triazole-azole bisheteroaryls. AB - The azido transfer procedure of heteroaryllithium and tosyl azide was used to synthesize selected 2- and 5-azidoazoles. This procedure, which is based on the fragmentation of the appropriate lithium triazene salts 1a-7a, successfully afforded 2-azido-N-methylimidazole 1, 2-azido-1,3-thiazole 2, 2-azidobenzo-1,3 thiazole 3, 5-azido-N-methylpyrazole 4, 5-azido-N-methylimidazole 6[via 2 (trimethylsilyl)-5-azido-N-methylimidazole 5], and 5-azido-1,3-thiazole 7 (via 5 lithio-1,3-thiazole), but attempts to prepare 5-azido-2-(trimethylsilyl)-1,3 thiazole 8 from the corresponding triazene 7a failed, affording only the desilylated azide in poor yield. Azides - underwent 1,3-dipolar cycloaddition when mixed with neat (trimethylsilyl)acetylene, giving 1-heteroaryl-4 trimethylsilyl-1,2,3-triazoles 1b-7b generally in very high yields. PMID- 15827650 TI - On the electronic effects of OH groups. Synthesis and investigation of tetrahydroxylated azabicycloheptanes. AB - Two stereoisomeric 2,3,5,6-tetrahydroxyazabicyclo[2.2.1]heptanes were synthesised and their base strengths determined. The 2,3,5,6-exo-isomer 1 and the 2,3-exo-5,6 endo-isomer 2 were prepared from the Diels-Alder adduct of Boc-pyrrole and tosylacetylene by a route involving osmium catalyzed dihydroxylation and protection, tosyl group reduction and repeated dihydroxylation. Deprotection gave 1, while 2 was obtained by conversion of the diol into the ditriflate, followed by nucleophilic inversion with KNO(2) and deprotection. Synthesis of the 2,3,5,6 endo-isomer by a similar strategy was attempted but failed. The pK(a) of 1 and 2 was determined to be 7.0 and 6.4 respectively. This means that the change in base strength as a result of stereoisomerism of an OH is smaller in the [2.2.1] azabicyclic system than in the piperidines. This is explained by a difference in charge-dipole interactions in the two systems. PMID- 15827651 TI - Development of new calcium receptors based on oxazolidin-2-ones containing pseudopeptides. AB - With the aim of designing a new calcium receptor, the synthesis and the conformational analysis of a small library of dipeptides having the general formula Ac-Oxx-L-Xaa-OBn [Oxx = L-Oxd, (4S,5R)-4-methyl-5-carboxyoxazolidin-2 one; D-Oxd, (4R,5S)-4-methyl-5-carboxyoxazolidin-2-one; or D-Oxac (4R)-(2-oxo-1,3 oxazolidin-4-yl)-acetic acid] is reported. Ac-L-Oxd-L-Ala-OBn was identified as the most promising compound by MS-ESI analysis and this outcome was confirmed by photoluminescence spectroscopy. PMID- 15827653 TI - Solvent-free condensation of phenylacetonitrile and nonanenitrile with 4 methoxybenzaldehyde: optimization and mechanistic studies. AB - The condensation of phenylacetonitrile with 4-methoxybenzaldehyde can be carried out by two solvent-free methods: i) using neat powdered KOH at room temperature with equivalent amounts of aldehyde, nitrile and base for 3 min, and ii) using K2CO3 in the presence of a phase transfer agent for 3 min under microwave activation or conventional heating. By extending the reaction time up to 10 min, four different products were obtained from phenyl or nitrile group migration. With nonanenitrile, only the second method could be applied to give two kinds of condensed compounds with or without phenyl (or nitrile) group migration. The intervention of non-thermal MW-specific effects was proved in some cases and interpreted. PMID- 15827652 TI - Mechanistic studies of La3+- and Zn2+-catalyzed methanolysis of aryl phosphate and phosphorothioate triesters. Development of artificial phosphotriesterase systems. AB - The methanolyses of a series of O,O-diethyl O-aryl phosphates (2,5) and O,O diethyl S-aryl phosphorothioates (6) promoted by methoxide and two metal ion systems, (La3+)2(-OCH3)2 and 4:Zn2+:-OCH3 (4 = 1,5,9-triazacyclododecane) has been studied in methanol at 25 degrees C. Bronsted plots of the logk2 values vs. pKa for the phenol leaving groups give beta(lg) values of -0.70, -1.43 and -1.12 for the methanolysis of the phosphates and -0.63, -0.87 and -0.74 for the methanolysis of the phosphorothioates promoted by the methoxide, La3+ and Zn2+ systems respectively. The kinetic data for the metal-catalyzed reactions are analyzed in terms of a common mechanism where there is extensive cleavage of the P-XAr bond in the rate-limiting transition state. The relevance of these findings to the mechanism of action of the phosphotriesterase enzyme is discussed. PMID- 15827654 TI - Towards multifunctional antioxidants: synthesis, electrochemistry, in vitro and cell culture evaluation of compounds with ligand/catalytic properties. AB - Numerous human diseases are linked to a biochemical condition known as oxidative stress (OS). Antioxidants are therefore becoming increasingly important as potential disease prevention and therapeutic agents. Since OS is a multi-stressor event, agents combining a range of different antioxidant properties, such as redox catalysis and metal binding, might be more effective and selective than mono-functional agents. Selenium derivatives of aniline and pyridine combine redox activity with metal binding properties. These multifunctional agents have a distinct electrochemical profile, and exhibit good catalytic activity in the glutathione peroxidase mimic and metallothionein assays. They also show antioxidant activity in a skin cell model of UVA-induced stress. These compounds might therefore provide the basis for novel agents combining two or more distinct antioxidant properties. PMID- 15827655 TI - Asymmetric hydrogenation using chiral Rh complexes immobilised with a new ion exchange strategy. AB - Rh diphosphine complexes using DuPhos and JosiPhos as chiral ligands have been immobilised by ion exchange into the mesoporous material MCM-41. When used as catalysts for the enantioselective hydrogenation of dimethyl itaconate and methyl 2-acetamidoacrylate, these heterogeneous catalysts give catalytic performance in terms of yield and enantioselection that are comparable to the corresponding homogeneous catalysts. Furthermore, the heterogeneous catalysts can be readily recovered and reused without loss of catalyst performance. A second immobilisation strategy is described in which [Rh(COD)2]+BF4- is initially immobilised by ion exchange and subsequently modified by the chiral diphosphine and this give comparable catalyst performance. This immobilisation strategy opens up the possibility of easy ligand-screening for parallel synthesis and libraries. PMID- 15827656 TI - Preparation of sialylated oligosaccharides employing recombinant trans-sialidase from Trypanosoma cruzi. AB - Terminally sialylated oligosaccharides were synthesised employing recombinant trans-sialidase from Trypanosoma cruzi. Regio- and stereoselectively Sia-alpha(2 3)-Gal-betaR derivatives could be obtained in respectable yields, using combined chemical and enzymatic methodologies. An array of different disaccharide precursors such as Gal-beta(1-3)-GalNAc-alphaSer/Thr, lactosides and lactosamide derivatives were sialylated and successfully purified by facile isolation procedures. Depending on the acceptor structure isolated, yields for trans sialylation products were between 20 and 60%. PMID- 15827657 TI - A synthesis of (+/-)-sparteine. AB - In a synthesis of racemic sparteine, Diels-Alder reaction between dimethyl bromomesaconate 14 and dicyclopentenyl 4, followed by cyclopropane formation, set up the stereochemistry at C-1 and C-5 as S and R, respectively, in a meso intermediate 8. The stereochemistry at C-2 and C-4 was then secured by a moderately diastereoselective protonation of the bis-enolate 17 derived from the diester 8 by reductive cleavage with lithium in liquid ammonia. The C=C in the racemic diester 19 was ozonolysed and the diketone converted by Beckmann rearrangement into the bis-lactam . Reduction of the bis-lactam with lithium aluminium hydride and intramolecular nucleophilic displacement gave racemic sparteine 1. Some ideas for making this synthesis amenable to a synthesis of enantiomerically enriched sparteine are presented. PMID- 15827658 TI - The formation and properties of the melatonin radical: a photolysis study of melatonin with 248 nm laser light. AB - The photolysis of melatonin in aqueous solution has been studied spectrometrically with a 248 nm laser. The formation of hydrated electrons in a monophotonic process has been confirmed in neutral solution with a quantum yield of 0.22. Two main absorption bands at 340 and 460 nm plus an absorption shoulder resulted from the counterpart of the ejected electron, a melatonin radical, in solution. The big difference for the relative intensity of the absorption peaks under various pH conditions reveals that the melatonin radical exists in the solution through an acid-base equilibrium. In support from the pH dependence of the spectrum of the intermediate, the pKa1 for the doubly-protonated melatonin radical against the mono-protonated melatonin cation radical was estimated to be 0.95 and the pKa2 for the mono-protonated melatonin cation and melatonin neutral radical was 4.5 +/- 0.5. This work will benefit the basic understanding about melatonin as a UV-light protector, as a light receptor and the antioxidation functions of melatonin. PMID- 15827659 TI - Empowering physicians and patients through greater knowledge of drugs, for safety's sake. PMID- 15827660 TI - Use of ligase chain reaction in early diagnosis of tuberculous meningitis. AB - INTRODUCTION: Nucleic acid-based amplification tests are currently licensed only for the detection of Mycobacterium tuberculosis in pulmonary specimens. There are insufficient data for extrapulmonary specimens. The aim of this study is to investigate the diagnostic value of these investigations in tuberculous meningitis. MATERIALS AND METHODS: We performed a prospective study using a commercial ligase chain reaction DNA amplification technique [Ligase chain result (LCx) M. tuberculosis; Abbott Laboratories, Abbott Park, IL, USA] on cerebrospinal fluid (CSF) to diagnose tuberculous meningitis and compared the results with standard microbiological data. Conflicting cases were resolved according to the final clinical diagnosis. A total of 54 CSF specimens from 54 patients were tested. RESULTS: Six (11.1%) specimens were culture-positive for M. tuberculosis; of these, only 1 (1.9%) was smear-positive. The sensitivity, specificity, positive predictive value and negative predictive value of the LCx assay, compared with culture results, were 66.7%, 100%, 100% and 96%, respectively. After resolution according to clinical data, the sensitivity, specificity, positive predictive value and negative predictive value were 33.3%, 100%, 100% and 75%, respectively. All controls had negative LCx results. There were no false positives. CONCLUSION: LCx assay is highly specific and complements conventional laboratory diagnostic methods (CSF smear and culture) in the diagnosis of tuberculous meningitis. In the appropriate clinical context, a positive result is strongly suggestive of tuberculosis and could enable antituberculous treatment to be started immediately. PMID- 15827661 TI - Factors associated with decompression and strabismus surgery in thyroid eye disease. AB - INTRODUCTION: Prognostication of the thyroid patient with eye disease aids in the choice of treatment strategy. To facilitate this, we investigated factors associated with decompression and/ or strabismus surgery in the Singaporean population. MATERIALS AND METHODS: A 5-year retrospective study was performed. Patients who required strabismus and/or decompression surgery (n = 23) were compared to those who did not undergo either surgery (n = 44). Individual and multivariate age-adjusted odds ratios were calculated to determine significant associations. RESULTS: Individually, male gender [odds ratio (OR), 4.5; 95% confidence interval (CI), 1.5 to 13.4], uncontrolled hyperthyroidism (OR, 4.0; 95% CI, 1.1 to 14.3), steroid therapy (OR, 7.4; 95% CI, 2.3 to 24), diplopia (OR, 7.3; 95% CI, 2.3 to 23.1), objective vertical myopathy (OR, 11.7; 95% CI, 1.4 to 96.0), elevated intraocular pressure in the primary position (OR, 3.4; 95% CI, 1.2 to 10.0) and clinical evidence of optic neuropathy (OR, 13.1; 95% CI, 1.4 to 124.6) were significantly associated with the need for surgery. Logistic regression analysis showed the greater impact of male gender (OR, 4.2; 95% CI, 1.2 to 15.4), optic neuropathy (OR, 13.0; 95% CI, 1.2 to 143.7) and previous steroid therapy (OR, 4.2; 95%CI, 1.1 to 16.2) on prognostication. CONCLUSIONS: Chances of requiring strabismus and/or decompression surgery are significantly higher for male patients and those with uncontrolled hyperthyroidism. In particular, male patients with optic neuropathy and a history of previous steroid therapy warrant a graver prognosis. PMID- 15827662 TI - Use of physical restraints in nursing homes: current practice in Singapore. AB - INTRODUCTION: One of the indicators of quality care in nursing homes is the proper use of physical restraints. Restraints have been associated with multiple complications, such as pressure ulcers, incontinence and immobility. The aim of our study was to assess the indication, use and complications related to physical restraints in Singapore nursing homes. MATERIALS AND METHODS: All residents aged >65 years old from 3 nursing homes were included in the study. A geriatrician reviewed the medical records of all elderly residents on restraints. Nursing staff interviews were also done to assess their knowledge regarding restraint use. A brief medical record review was also done for the elderly residents without restraints. RESULTS: Out of 390 elderly residents, 91 were on physical restraints. The mean age of these residents were 80.1 years; 67% were female, 90.1% were Chinese and 82.4% belonged to functional category IV. Both urinary and faecal incontinence were observed in 97.8% of residents. Forty-six (50.5%) residents had no documented indication for restraints. It was noted that 20 (22%) residents were restrained to prevent dislodgement of the feeding tube, 17 (18.7%) were restrained to prevent falls and 8 (8.8%) were restrained for agitation. There were documented trials of removal of restraints for only 21 (23.1%) residents. Alternative approaches, such as diversion technique, were used for 31 (34.1%) residents to avoid restraint use. CONCLUSION: Our study revealed a high rate of restraint use in the nursing homes. A restraint protocol should be available in every nursing home, requiring physician approval for the long-term application of physical restraints and intermittent reviews by physicians to assess the continued need for restraints. PMID- 15827663 TI - The relationship between postoperative complications and outcomes after hip fracture surgery. AB - INTRODUCTION: We studied the prevalence of postoperative complications in a series of consecutive patients who received surgery for hip fractures in a major public hospital in Singapore. We also studied the predictors for the occurrence of complications and the impact of these complications on patient outcomes. MATERIALS AND METHODS: A retrospective chart review of patients admitted with hip fracture, from March to November 2001, was carried out. Patients were classified as having postoperative complications if they developed any of the following conditions after surgery: dislocation of prosthesis, deep vein thrombosis, postoperative confusion, foot drop, stroke, cardiac arrhythmias or acute myocardial infarctions, urinary retention, urinary tract infection, pneumonia, wound infection and incident pressure sores. RESULTS: Of the 180 patients studied, 60 developed postoperative complications. Significant predictors of complications after logistic regression included being of female gender [odds ratio (OR), 2.79; 95% confidence interval (CI), 1.13 to 6.89] and pre-fracture mobility status (OR for independent ambulators 0.45; 95% CI, 0.23 to 0.87), but not the age of the patients. Postoperative complications significantly affected the length of stay within the acute hospital (beta coefficient, 6.42; 95% CI, 2.55 to 10.29), but were not associated with a decline in mobility status at 3 months post-fracture, eventual discharge destination or readmission within 1 year. CONCLUSION: Postoperative complications are common after surgery for hip fractures and result in significantly longer hospitalisation periods. Significant predictors for such complications include being of female gender and pre-fracture mobility. Age, in itself, does not result in a higher risk of complications and should not preclude older hip fracture patients from surgical management. PMID- 15827664 TI - Incidence, risk factors of retinopathy of prematurity among very low birth weight infants in Singapore. AB - INTRODUCTION: To determine the incidence, risk factors and need for surgery for retinopathy of prematurity (ROP) among very-low-birth-weight (VLBW) infants. MATERIALS AND METHODS: This was a retrospective study of all VLBW infants managed by the department over 14 years, from 1988 to 2001. Preterm infants were examined according to the Royal College of Ophthalmologists' guidelines, and retinopathy was graded following the International Classification of ROP. All VLBW infants examined for ROP were included and data were retrieved retrospectively and analysed for maternal, medical, obstetric and neonatal risk factors using logistic regression. RESULTS: Of the 564 VLBW infants who fit the screening criteria, ROP was detected in 165 (29.2%) of VLBW infants; of whom 49% of infants had stage 1 disease, 24% were at stage 2, and 27% were at stage 3 or more. Among 45 infants with stage 3 disease or more, treatment was needed in 62.2% (28/45). No ROP was detected in infants greater than 33 weeks of gestation. Only 0.6 % (1/164) of infants greater than 30 weeks of gestational age (GA) needed surgery for ROP. Using birth weight (BW) criteria, stage 3 ROP was noted only in 1% (6/564) of infants with BW >1000 g. Of all ROP requiring surgery, 89% (25/28) of infants were <1000 g as compared to 11% (3/28) who were >1000 g infants. The median age of onset of ROP was 35 weeks (range, 31 to 41) corrected age. By univariate analysis for threshold ROP, preeclampsia, prenatal betamethasone exposure, gestational age, birth weight, 1-minute Apgar score, hyaline membrane disease (HMD), surfactant usage, hypotension, septicaemia, intraventricular haemorrhage duration of supplemental oxygen, ventilation and chronic lung disease were associated with ROP requiring surgery (i.e., threshold ROP, P <0.05). However, using multiple logistic regression analyses for ROP, maternal preeclampsia [odds ratio (OR), 2.52; confidence interval (CI), 1.32 to 4.7], birth weight (OR, 0.99; CI, 0.996 to 0.999), pulmonary haemorrhage (OR, 4.61; CI, 1.04 to 20.4), duration of ventilation (OR, 1.06; CI, 1.04 to 1.08) and duration of continuous positive airway pressure (CPAP) (OR, 1.02; CI, 1.01 to 1.04) were factors predictive of development of threshold ROP. CONCLUSION: The incidence of ROP among VLBW infants was 29.2%. ROP was strongly associated with smaller, more immature and sicker infants. The median age of onset of ROP was 35 weeks (range, 31 to 40 weeks) postmenstrual age. Infants <30 weeks of GA and/or infant with BW <1000 g are at considerable risk for threshold ROP. The main risk factors for development of threshold ROP by regression analysis are maternal preeclampsia, birth weight, and presence of pulmonary haemorrhage, duration of ventilation and continuous positive pressure ventilation. We suggest that both immaturity and compromised pulmonary function are both important aetiological factors in the development of ROP. Prevention of prematurity, control of preeclampsia, judicious use of ventilation and oxygen therapy are the only promising factors that may reduce the incidence and severity of ROP in this high-risk infant. PMID- 15827665 TI - Contemporary results of endopyelotomy for ureteropelvic junction obstruction. AB - INTRODUCTION: Endopyelotomy is an accepted treatment option for ureteropelvic junction obstruction (UPJO). In this study, we reviewed our 7-year experience with antegrade endopyelotomy for UPJO. MATERIALS AND METHODS: We reviewed the records of 35 consecutive antegrade endopyelotomy for UPJO between 1996 and 2002. Patients were included if they had shown radiographic evidence of UPJO on diuresis urography or intravenous urogram with signs and symptoms or deterioration of renal function. RESULTS: A total of 34 consecutive patients underwent 35 antegrade endopyelotomy procedures in 35 renal units. One patient had bilateral endopyelotomy for bilateral UPJO. Eighteen renal units (51%) had concomitant renal calculi that required percutaneous nephrolithotomy, including 8 renal units with pelvi-ureteric junction stones. Twenty-four renal units (69%) had moderate degree of hydronephrosis whilst 11 renal units (31%) had severe hydronephrosis. The mean operating time for antegrade endopyelotomy was 94 +/- 28 minutes and the mean hospital stay was 4.7 +/- 2.8 days. No patients had conversion to open pyeloplasty and no patient required perioperative blood transfusion. The mean followup was 33 +/- 23 months and the overall success rate following endopyelotomy was 83% (n = 29 renal units). The success rate for primary UPJO was 81%, whilst the success rate for secondary UPJO was 84%. Four renal units (11%) required ancillary procedures for failed endopyelotomy. Two patients required repeat endopyelotomy, and 2 patients needed open pyeloplasty. Two patients were lost to follow-up. CONCLUSION: Endopyelotomy remains a viable approach for UPJO compared to open reconstruction. Careful patient selection can optimise the surgical outcome and minimise endopyelotomy failures. PMID- 15827666 TI - Risk perception is affected by modes of risk presentation among Singaporeans. AB - INTRODUCTION: Not much is known about how Singaporeans perceive and react to risk presentation. There is no consensus on whether the European Union guidelines for describing the risk of side effects are valid. This study investigated the effect of different modes of risk presentation on how Singaporeans perceive and react to medical risk. Furthermore, we investigated the practical usage of qualitative phrases, the European Union's adjectives in particular, in describing levels of risk. MATERIALS AND METHODS: A hypothetical situation about the risk of side effects of an influenza vaccine was presented in either a probability format (i.e., 5%; n = 42) or a frequency format (i.e., 1 out of 20; n = 43). The 2 versions of questionnaire were handed out in an alternate order to a convenience sample of 47 healthcare professionals and 38 university students. RESULTS: Respondents presented with a "5% risk" were more likely to describe the risk as "uncommon" or "rare", as compared to respondents presented with a risk of "one out of twenty" (P <0.01). Furthermore, the former showed more willingness to accept the influenza vaccine described in the hypothetical situation than in the latter, but this was not statistically different (67% versus 54%; P >0.1). CONCLUSIONS: Modes of risk presentation affect how people perceive risk, even among people who are highly educated. PMID- 15827667 TI - A walk through the management of Parkinson s disease. AB - INTRODUCTION: Patients with Parkinson s disease are known to develop motor complications after a few years of therapy. Motor fluctuations and dyskinesias develop with increasing severity of disease, and were formerly thought to be an inevitable consequence of the disease. METHODS: Literature review of articles on the aetiopathogenesis of Parkinson s disease, the mechanisms underlying the development of motor fluctuations and dyskinesias, and strategies for delaying the onset of dyskinesias. RESULT: Motor fluctuations develop with increasing severity of the disease, owing to loss of dopaminergic neurons and loss of the buffering capacity of the neurons to fluctuating dopamine levels. Dyskinesias develop as a result of pulsatile stimulation of the receptors, causing changes in plasticity, dysregulation in gene and protein expression and alterations in neuronal firing patterns. Continuous dopaminergic stimulation, through long acting dopa agonists and frequent administration of levodopa, is known to delay the development of dyskinesias. The use of catechol-O-methyl transferase (COMT) inhibitors likewise increases the bioavailability and brings about a smooth drug profile. The use of dopa agonists is associated with sedation and confusion, particularly in the elderly. CONCLUSIONS: Initiation of therapy in Parkinson s disease should begin with a dopa agonist agent, unless the patient is elderly or has cognitive impairment, in which case levodopa therapy should be given. PMID- 15827668 TI - Multimodal antiemetic therapy and emetic risk profiling. AB - INTRODUCTION: Postoperative nausea and vomiting (PONV) is a common problem with no simple solution. This review highlights factors that are known to increase the risk of PONV. It examines the various data on pharmacological and non pharmacological methods that have been used to prevent PONV. METHODS: Peer reviewed journals on the subject were covered. CONCLUSION: Patient, surgical and anaesthetic factors increase the risk of PONV. While patient and surgical factors are understandably difficult to control, a multimodal approach involving both pharmacological and non-pharmacological interventions has been successfully adopted to reduce the incidence of PONV. Various factors have been identified to categorise patients into different profiles to determine their risk of PONV. Perioperative strategies can then be targeted at these patient groups. PMID- 15827669 TI - Mesenchymal stem cells in musculoskeletal tissue engineering: a review of recent advances in National University of Singapore. AB - A key factor in the tissue engineering approach to tissue repair and regeneration is the use of appropriate cells. Mesenchymal stem cells (MSCs) are derived from bone marrow stroma or connective tissues and they have the potential to differentiate into various mesenchymal cell lines in vitro and in vivo. These cells hold great promise for musculoskeletal tissue engineering. This review is based mainly on the work which has been done in the National University of Singapore on the use of MSCs for engineering cartilage, growth plate, bone and tendon/ligament as well as the clinical trail of autologous chondrocyte implantation. It can help to shape future research on musculoskeletal tissue engineering. PMID- 15827670 TI - Tuberculosis post-liver transplantation: a rare but complicated disease. AB - INTRODUCTION: Tuberculosis is a rare but serious complication after transplantation. We report a case and discuss its presentation and management. CLINICAL PICTURE: A 60-year-old Indonesian male presented initially with fever, acute confusion and rapidly progressive right upper lobe pneumonia 3.5 months post-liver transplant, and was diagnosed with pulmonary tuberculosis by positive sputum smear for acid-fast bacilli and tuberculosis culture. TREATMENT/OUTCOME: Standard anti-tuberculosis therapy was administered but was complicated by interaction with cyclosporine and drug-induced cholestasis. CONCLUSION: A high level of suspicion, prompt antituberculosis treatment and close follow-up are essential in management of post-transplant tuberculosis. PMID- 15827671 TI - Caesarean scar pregnancy: a diagnosis to consider carefully in patients with risk factors. AB - INTRODUCTION: Caesarean scar pregnancy is rare. A high index of suspicion is necessary to make the right diagnosis early. CLINICAL PICTURE: Two women, each with a history of 1 previous caesarean delivery, were initially diagnosed with spontaneous abortion in progress when ultrasonography scan revealed the gestational sac to be in the lower uterine cavity. Subsequent ultrasound scans showed that the gestational sac was actually in the anterior uterine wall at the level of the uterine isthmus. TREATMENT: Both women were managed successfully with intrasac methotrexate injection. OUTCOME: We report 2 cases of caesarean scar pregnancies mistaken for spontaneous abortions by experienced ultrasonographers. CONCLUSION: This highlights the difficulty in differentiating between these 2 diagnoses and the importance of having a high index of suspicion in women with risk factors. PMID- 15827672 TI - Economic terms of surgical practice. PMID- 15827673 TI - Nomograms and staging in melanoma: new tools for better predicting outcomes. PMID- 15827674 TI - Surgically resected hepatocellular carcinoma: protean yet predictable. PMID- 15827675 TI - Osteosarcoma of the hand and forearm: experience of the Cooperative Osteosarcoma Study Group. AB - BACKGROUND: Osteosarcoma is extremely rare in the hand and forearm. Therefore, only limited data are available for planning treatment or predicting the outcome and prognosis of osteosarcoma in the peripheral upper extremity. METHODS: Epidemiological, clinical, and histopathologic data were analyzed in 39 patients with osteosarcoma of the forearm or hand who were enrolled in studies of the Cooperative German-Austrian-Swiss Osteosarcoma Study Group from 1977 to December 2000. In patients with high-grade osteosarcoma, the treatment entailed surgical resection in combination with chemotherapy, whereas patients with low-grade osteosarcoma underwent only surgery. RESULTS: The 5-year overall survival rate among the 33 patients with high-grade central osteosarcoma of the distal upper extremity was 86.2% +/- 6.4%. The 5-year event-free survival rate was 65.4% +/- 9.6%. Five of the eight patients with secondary metastases were in remission at the time of analysis. Four patients died of their disease, and two patients died of chemotherapy-related complications. The mean overall survival rate was 88.0% +/- 6.5% in patients treated by wide or radical tumor resection and was 75.0% +/- 21.7% in patients with nonwide margins of resection. Whether amputation or local resection was performed had no significant influence on the prognosis. All six patients whose osteosarcoma was not classified as high-grade central osteosarcoma were in remission at the time of analysis. CONCLUSIONS: The results demonstrate a remarkably high survival rate for patients with high-grade osteosarcoma of the hand and forearm and confirm that multiagent chemotherapy in combination with wide excision is a highly effective treatment for this malignant tumor. PMID- 15827676 TI - RON, a tyrosine kinase receptor involved in tumor progression and metastasis. AB - Tyrosine kinase receptors mediate many critical cellular functions that contribute to tumor progression and metastasis and thus are potential targets for molecular-based cancer therapy. As has been found for many receptor tyrosine kinases, RON (recepteur d'origine nantais) and its ligand, macrophage-stimulating protein, have recently been implicated in the progression and metastasis of tumors. In in vitro experiments using colon and breast cancer cell lines, overexpression of RON led to increased invasion and migration of cancer cells and prevented apoptosis and anoikis. In addition, transgenic mice engineered to overexpress RON in the lung epithelium developed multiple pulmonary tumors, suggesting a role for RON in tumorigenesis. In human cancer specimens, increased RON expression has been demonstrated in colon, breast, ovarian, and lung tumors. Therefore, therapies designed to inhibit RON activation may hinder critical tumor survival mechanisms and play a role in the treatment of advanced disease. PMID- 15827677 TI - A practical approach to intraoperative evaluation of sentinel lymph node biopsy in breast carcinoma and review of the current methods. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy is increasingly becoming an alternative method for assessing axillary status in breast carcinoma patients. Intraoperative SLN evaluation can potentially select patients for immediate axillary clearance and spare most of them a second surgical procedure. Nevertheless, no standard protocol for intraoperative SLN evaluation has been developed. The aims of this study were to establish the reliability of SLN intraoperative evaluation in breast carcinoma staging, to review the published methods currently used, and to propose a standard protocol. METHODS: One hundred fifty-two SLNs were collected from 86 patients. Lymphoscintigraphy, blue dye, and gamma camera intraoperative controls were used for localization. Each SLN was sliced 2 mm thick and was intraoperatively evaluated by using the combination of frozen section and imprint cytology. The final examination included standard hematoxylin and eosin staining, and, in case of persistent negativity, further sectioning, including hematoxylin and eosin combined with immunohistochemistry (CAM5.2 cytokeratin), was performed. RESULTS: The combination of frozen section and imprint cytology for intraoperative SLN evaluation yielded an intraoperative sensitivity of 78% and a specificity of 100%. All macrometastases (>2 mm) were detected during surgery, as were 2 micrometastases. Final examination detected seven more micrometastases, six of which consisted of isolated tumor cells. CONCLUSIONS: We propose a fast, cost-effective, and accurate procedure for SLN evaluation that is useful for making intraoperative decisions, feasible for most institutions, and reliable because of its high sensitivity (100% for macrometastases) and specificity. PMID- 15827678 TI - Results of hysterectomy in patients with bulky residual disease at the end of chemoradiotherapy for stage IB2/II cervical carcinoma. AB - BACKGROUND: We assessed the clinical outcome after hysterectomy in patients with bulky residual disease after chemoradiotherapy for stage IB2/II cervical carcinoma. METHODS: Subjects were 10 patients who had bulky (>2 cm) residual disease in the cervix after external radiotherapy (45 Gy) combined with concomitant chemotherapy (cisplatin 40 mg/m2/week) and uterovaginal brachytherapy (15 Gy). RESULTS: Extrafascial hysterectomy was performed in three patients, type II radical hysterectomy was performed in six patients, and pelvic exenteration was performed in one patient. Pelvic lymphadenectomy was performed in eight patients, and para-aortic lymphadenectomy was performed in eight. Five patients had nodal involvement (pelvic nodes in four and para-aortic nodes in four), and six had lymphovascular space involvement. Surgical margins were free in nine patients. Seven patients developed grade 2 (n = 3) and/or grade 3 (n = 4) complications. The median duration of follow-up after surgery was 22 months (range, 1-37 months). With follow-up available in nine patients, seven relapsed, and only two remained disease free. CONCLUSIONS: This series confirms the high rate of nodal spread in patients with bulky residual cervical disease after chemoradiotherapy. Furthermore, patients who underwent hysterectomy had a high complication rate. Only two patients are alive and disease free. The results of surgery are disappointing; surgery does not seem to improve the survival of these patients. PMID- 15827679 TI - A nomogram that predicts the presence of sentinel node metastasis in melanoma with better discrimination than the American Joint Committee on Cancer staging system. AB - BACKGROUND: The threshold and indications for sentinel lymph node (SLN) biopsy in patients with melanoma remain somewhat arbitrary. Many variables associated with SLN positivity have previously been identified, including a significant association between the American Joint Committee on Cancer (AJCC) staging system and SLN status. We developed a user-friendly nomogram that takes several characteristics into account simultaneously to more accurately predict the presence of SLN metastasis for an individual patient. METHODS: A total of 979 patients who underwent successful SLN biopsy for cutaneous melanoma at a single institution between February 1991 and November 2003 were included in the analysis. Predictors were used to develop a nomogram, based on logistic regression analysis, to predict the probability of SLN positivity. A large multi institutional trial with 3108 patients was used to validate the predictive accuracy of the nomogram compared with the AJCC staging system. RESULTS: The nomogram was developed and found to be accurate and discriminating. The concordance index of the nomogram, a measure of predictive ability, was .694 when evaluated with the validation dataset. In contrast, the concordance index of the AJCC staging system was lower (.663; P < .001). CONCLUSIONS: Using commonly available clinicopathologic information, we developed a nomogram to accurately predict the probability of a positive SLN in patients with melanoma. This tool takes several characteristics into account simultaneously. This model should enable improved patient counseling and treatment selection. PMID- 15827680 TI - Arteriolovenular shunting critically determines shutdown of microcirculation upon cryotherapy in tumor-bearing rat liver. AB - BACKGROUND: Tissue destruction by cryosurgery not only is mediated by direct cell damage, but also involves secondary mechanisms, such as ischemia due to shutdown of the microcirculation. Clinicians favor repetitive cryoapplication, although there is no proven evidence for a more effective tumor eradication. METHODS: The aims of this study were (1) to establish a rat liver tumor model that allows for intravital microscopic analysis of hepatic tumor microcirculation and (2) to elucidate critical determinants of shutdown of microvascular perfusion after single and repetitive cryotherapy. In WAG-Rji rats (n = 14), syngeneic colon carcinoma cells (CC531) were implanted into the left liver lobe. Hepatic and tumor microcirculation were studied by intravital microscopy. RESULTS: Two weeks after implantation, the tumors had developed a microvasculature with a capillary density markedly (P < .05) lower compared with the sinusoidal density of normal liver. However, at the tumor margin, venule diameters were significantly enlarged (P < .05), with high red blood cell velocities and arteriolovenular shunts. Both freeze procedures (temperature at the tumor margin: -32.4 degrees C +/- 1.6 degrees C and -36.4 degrees C +/- 2.0 degrees C) resulted in a complete shutdown of intratumoral and peritumoral capillary and hepatic sinusoidal perfusion. In contrast, some large venules showed maintenance of blood flow initially after freezing (15 minutes); however, this was abolished during the subsequent 2-hour observation period. CONCLUSIONS: Enlarged high-flow venules at the tumor margin, which participate in arteriolovenular shunting, critically determine the shutdown of the microcirculation upon cryotherapy. Repetitive freezing is not more effective than a single-freeze procedure to achieve complete tumor microcirculatory stasis. PMID- 15827681 TI - Prognostic evaluation of the new American Joint Committee on Cancer/International Union Against Cancer staging system for hepatocellular carcinoma: analysis of 112 cirrhotic patients resected for hepatocellular carcinoma. AB - BACKGROUND: In 2002, the American Joint Committee on Cancer and the International Union Against Cancer redefined the T-classification for hepatocellular carcinoma, shifting the cutoff value for tumor size from 2 to 5 cm and giving more emphasis to vascular invasion. METHODS: A retrospective cohort study was conducted on 223 consecutive patients with hepatocellular carcinoma observed between 1990 and 2002. One hundred twelve were resected and considered for retrospective analysis. Univariate and multivariate analyses were performed on several clinicopathologic variables. After classification according to each staging system, the long-term survival of different stages was compared. The prognostic value of each staging system was further evaluated by entering each stage, in turn, into the Cox regression model with other clinicopathologic variables. The median follow-up was 19 months. RESULTS: On multivariate analysis, the viral etiology of cirrhosis and the presence of multiple nodules were independent prognostic factors. When the new staging system was entered into the multivariate analysis, it was the only independent factor (P = .02). When stratified according to the old tumor-node metastasis system, there were no significant differences in the survival between stage I and II (P = .14) or between stage IIIA and IVA (P = .33); only the survival of stage II and IIIA was different (P < .01). When stratified according to the new tumor-node-metastasis system, there were significant differences between stage I and II (71.7% vs. 54.7%; P = .02). CONCLUSIONS: The new staging system is a more reliable and objective method for T classification. It is easy to use in clinical practice and is better at stratifying curatively resected patients with respect to prognosis. PMID- 15827682 TI - Incidence of unsuspected and treatable metastatic disease associated with operable colorectal liver metastases discovered only at laparotomy (and not treated when performing percutaneous radiofrequency ablation). AB - BACKGROUND: When patients with resectable colorectal liver metastases (LM) are treated with percutaneous radiofrequency (RF), some unsuspected intrahepatic and extrahepatic metastases, detectable only at laparotomy, might be ignored and left untreated. This would result in a reduced cure rate. Our purpose was to discover the incidence of unsuspected and surgically treatable intrahepatic and extrahepatic metastases discovered at laparotomy. METHODS: The data of 506 patients who underwent a laparotomy and then a hepatectomy for colorectal LM were prospectively collected and retrospectively analyzed. All patients had undergone at least two types of preoperative liver imaging (but no fluorodeoxyglucose positron emission tomography). RESULTS: Unsuspected metastases were discovered at laparotomy in 209 patients (41.3%). There were extrahepatic metastases in 82 patients (16.2%), additional LM in 152 patients (30%), and both in 25 patients (4.9%). Liver palpation and intraoperative ultrasound allowed for detecting additional LM in 125 (24.7%) and 48 (9.4%) patients, respectively. All of them were resected. When only the 124 patients who presented with 1 to 3 LM measuring <3 cm in diameter (candidates for percutaneous RF) were considered, the results were similar. Moreover, the incidence of unsuspected metastases was similar when the periods of surgery (before and after January 1996) were considered. CONCLUSIONS: Laparotomy permits discovery of and treatment with a curative intent of unsuspected intrahepatic or extrahepatic metastases in at least one third of patients with classically resectable colorectal LM. This does not support the use of percutaneous RF ablation instead of hepatic resection for this population, because it will result in an important survival decrease. PMID- 15827683 TI - Interactive effects of polyphenols, tocopherol and ascorbic acid on the Cu2+ mediated oxidative modification of human low density lipoproteins. AB - BACKGROUND: Only limited knowledge is available about any interactions between phenolic compounds and other antioxidants in inhibiting LDL oxidation. Many foods and beverages contain high levels of phenolic compounds; therefore, these compounds should not be considered in isolation from each other. AIM OF THE STUDY: The aim of this study was to examine the structure-antioxidant activity relationship of quercetin, caffeic acid, epicatechin, hesperetin and phloretin as well as alpha-tocopherol and ascorbic acid through their ability to interact with copper ions. METHODS: Isolated human LDL were incubated with single antioxidants or a combination of two and the kinetics of lipid peroxidation were assessed by measurement of conjugated diene formation (lag phase) via monitoring the absorbance at 234 nm after addition of copper ions. In addition, the degree of oxidation of the LDL protein moiety was followed by tryptophan fluorescence and carbonyl content measurements. RESULTS: Alpha-tocopherol and ascorbic acid showed a lower antioxidant activity in all test systems as compared to polyphenols at equimolar concentrations. Quercetin was the most effective compound in all three systems (p < 0.001 for lag phase and carbonyl content determination). A significant (p < 0.001) prolongation of the lag phase was found when combinations of ascorbic acid/quercetin, ascorbic acid/epicatechin, epicatechin/caffeic acid, and quercetin/epicatechin were tested as compared to the sum of the individual effects. Concerning the effects on LDL protein oxidation, the results from carbonyl content and the tryptophan fluorescence measurements showed that the combination of quercetin and caffeic acid revealed the strongest inhibitory effect (p < 0.001 carbonyl content; p < or = 0.002 tryptohan fluorescence) on protein oxidation which was higher than the effect of the single compounds. CONCLUSIONS: The results of the present study indicate that a combination of different antioxidants can be superior to the action of single antioxidants in protecting LDL lipid and protein moiety against oxidation. However, the substances may act by different antioxidative mechanisms, which are not necessarily complementary. PMID- 15827684 TI - Massive Gorham-Stout syndrome of the pelvis. AB - Gorham-Stout disease is defined as a spontaneous, massive, and nonfamilial idiopathic osteolysis. The diagnosis is based essentially on radiological and histological findings. Biopsy reviews always reveal excessive intraosseous nonmalignant proliferation of small vessels, which results in bone resorption and may extend to adjacent bones and soft tissues. These lesions are progressively replaced by extensive fibrosis. Since its first description in 1955, there is still controversy about its prognosis, etiology, and treatment. A case of Gorham Stout disease, located on the right pelvis, is reported with 50 years of clinical and radiographic follow-up, in a man who has never been treated. To date, this is the longest documented case report of the disease and its rare natural history. It demonstrates that after a variable time of evolution, the massive osteolysis is able to undergo spontaneous arrest and that the lesions may remain stable during several decades. Besides, no reossification was observed, even after 37 years of disease quiescence. Based on a large review of the literature, the authors then discuss the prognosis, etiopathology, and different therapeutic options available to halt the progression of the osteolysis. PMID- 15827685 TI - Osteopoikilosis in an ancient skeleton: more than a medical curiosity. AB - We describe the palaeopathologic and radiographic findings of the human skeletal remains that belonged to a female who lived in Mexico's viceroyship period (seventeenth and eighteenth centuries A.D.). Radiographic studies showed numerous, radiodense, ovoid, small and well-defined foci in the long tubular bones, sacrum, scapulae and iliac bones. Computed tomography (CT) examination revealed multiple hyperdense foci located in the central marrow portion of the bones. Measurements of attenuation coefficient revealed +1548 HU. The findings are consistent with osteopoikilosis, an uncommon, benign sclerosing bone dysplasia transmitted in an autosomal dominant fashion, which in the clinical setting is important to set apart from different bone pathologies to avoid unnecessary interventions and treatments. To the best of our knowledge, this is the first report of osteopoikilosis in ancient human remains. PMID- 15827687 TI - Determination of asymmetric and symmetric dimethylarginines in plasma of hyperhomocysteinemic subjects. AB - The aim of this study was to investigate the possible relationship among dimethylarginines (asymmetric, ADMA; symmetric, SDMA) and homocysteine (Hcy) levels in subjects affected by chronic, mild to intermediate, hyperhomocysteinemia. ADMA and SDMA were assayed by an optimised HPLC method in 75 patients (Hcy = 20.8 micromol/L, 17.1-30.2; median and percentile range) and, for comparison, in 85 healthy subjects (Hcy = 8.0 micromol/L, 7.0-9.1). In controls, the cut-off values were set at 0.61 micromol/L for ADMA and 0.56 or 0.48 micromol/L for male and female SDMA, respectively. In patients, ADMA and SDMA levels were increased (p < 0.001) with respect to controls, but no correlation with Hcy was observed. Hyperhomocysteinemic subjects showed a different behaviour in respect to ADMA and SDMA levels and this allowed their stratification in 3 subgroups characterized by ADMA and SDMA in the normal range, only SDMA, or both ADMA and SDMA over the cut-off values. A lack of correlation with Hcy was again observed, thus minimizing the direct role of Hcy on ADMA and SDMA metabolism and suggesting the need for further studies on this issue. PMID- 15827688 TI - Liberation of amino acids by heterotrophic nitrogen fixing bacteria. AB - Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed. PMID- 15827686 TI - Myelodysplastic syndrome associated with multiple autoimmune disorders. AB - The association between myelodysplastic syndromes (MDS) and autoimmune manifestations is not uncommon. As a rule, autoimmune abnormalities follow the diagnosis of MDS. We describe here a patient with MDS who developed a striking spectrum of diverse autoimmune disorders, including dermatitis, polyarthritis, and vasculitis, which preceded the clinical appearance of MDS. PMID- 15827689 TI - Glass transition of the two distinct single-chain particles of poly(N isopropylacrylamide). AB - Two distinct single-chain particles of poly(N-isopropylacrylamide) (PNIPAM) in the state of loose coil and compact globule, have been prepared successfully below and above the lower critical solution temperature (LCST) in extreme dilute aqueous solution by the freeze-drying method, respectively. During the preparation of the compact globular single-chain sample, the surfactant of sodium n-dodecyl sulfate (SDS) was added into the system to prevent aggregation of globular single chains formed at a temperature above the LCST. After all the coil has been transformed into the compact globular particle, the SDS molecules were removed by dialysis. The glass transition temperature (T(g)) of the two single chain samples has been measured by differential scanning calorimetery (DSC) in comparison with that of bulk polymer. It was found that the T(g) of the single chain sample in compact-globule state was very near to that of the bulk polymer, whereas the T(g) of the single-chain sample in loose-coil state was approximately 6 K lower than that of the bulk polymer. After treating the sample with repeated DSC cycles, the T(g) of the single-chain sample in loose-coil state rose up successively near to that of the bulk polymer. These results have been explained in terms of the effect of entanglement on the mobility of the polymer segments in the two distinct single-chain samples. PMID- 15827690 TI - Combined-modality treatment and organ preservation in bladder cancer. Do molecular markers predict outcome? AB - PURPOSE: In invasive bladder cancer, several groups have reported the value of organ preservation by a combined-treatment approach, including transurethral resection (TUR-BT) and radiochemotherapy (RCT). As more experience is acquired with this organ- sparing treatment, patient selection needs to be optimized. Clinical factors are limited in their potential to identify patients most likely to respond to RCT, thus, additional molecular markers for predicting treatment response of individual lesions are sorely needed. PATIENTS AND METHODS: The apoptotic index (AI) and Ki-67 index were evaluated by immunohistochemistry on pretreatment biopsies of 134 patients treated for bladder cancer by TUR-BT and RCT. Expression of each marker as well as clinicopathologic factors were then correlated with initial response, local control and cancer-specific survival with preserved bladder in univariate and multivariate analysis. RESULTS: The median AI for all patients was 1.5% (range 0.2-7.4%). The percentage of Ki-67-positive cells in the tumors ranged from 0.2% to 85% with a median of 14.2%. A significant correlation was found for AI and tumor differentiation (G1/2: AI = 1.3% vs. G3/4: AI = 1.6%; p = 0.01). A complete response at restaging TUR-BT was achieved in 76% of patients. Factors predictive of complete response included T-category (p < 0.0001), resection status (p = 0.02), lymphovascular invasion (p = 0.01), and Ki 67 index (p = 0.02). For local control, AI (p = 0.04) and Ki-67 index (p = 0.05) as well as T-category (p = 0.005), R-status (p = 0.05), and lymphatic vessel invasion (p = 0.05) reached statistical significance. Out of the molecular markers only high Ki-67 levels were associated to cause-specific survival with preserved bladder. On multivariate analysis, T-category was the strongest independent factor for initial response, local control and cancer-specific survival with preserved bladder. CONCLUSION: The indices of pretreatment apoptosis and Ki-67 predict a favorable outcome in bladder cancer patients treated with TUR-BT and RCT. Molecular markers may help to select patients for an organ-sparing approach. PMID- 15827691 TI - Induction chemotherapy with cisplatin, epirubicin, and paclitaxel (CEP), followed by concomitant radiotherapy and weekly paclitaxel for the management of locally advanced nasopharyngeal carcinoma. A Hellenic Cooperative Oncology Group phase II study. AB - BACKGROUND: Clinical research on the treatment of nasopharyngeal cancer (NPC) has been focused primarily on the reduction of incidence of the development of distant metastases as well as the improvement of locoregional control. PATIENTS AND METHODS: Untreated patients with stage IIB-IVB nonmetastatic NPC were treated with three cycles of induction chemotherapy (IC) consisting of epirubicin 75 mg/m(2) followed by paclitaxel 175 mg/m(2) as 3-h infusion on day 1 and cisplatin 75 mg/m(2) on day 2 every 3 weeks, followed by concomitant radiation therapy (70 Gy), and chemotherapy (CCRT) with weekly paclitaxel 60 mg/m(2). RESULTS: From November 1999 until April 2003, 47 patients entered the study. Complete response rate post IC therapy was 15%, which was raised to 66% after the completion of CCRT. The most frequent side effect from IC was myelotoxicity (55%), whereas stomatitis and xerostomia were the most pronounced (grade 3, 4) toxicities during CCRT. The presence of Epstein-Barr virus (EBV) was detected either by in situ hybridization in tumor tissue sections or by polymerase chain reaction in the peripheral blood in 37 out of 46 patients tested (80%). All three histological types were associated with the presence of EBV. After a median follow-up of 23.5 months, median time to treatment failure was 17.9 months, whilst median survival has not been reached yet. CONCLUSION: IC followed by CCRT is feasible and produces durable complete responses in the majority of patients with NPC. The case detection rate of EBV in this study appears to be similar to that reported from endemically infected regions. PMID- 15827692 TI - Different saliva substitutes for treatment of xerostomia following radiotherapy. A prospective crossover study. AB - BACKGROUND AND PURPOSE: Xerostomia is an important chronic side effect of radiotherapy in the head and neck area. The authors investigated the efficacy of different artificial saliva compounds in patients with postirradiation xerostomia. PATIENTS AND METHODS: In 120 patients with xerostomia after radiotherapy for head and neck cancer, four different saliva substitute compounds (gel, carmellose spray, oil, mucin spray) were tested in a prospective crossover design. Xerostomia at baseline and under treatment with each compound was measured with a questionnaire approved in a pilot trial. RESULTS: All compounds significantly improved xerostomia when compared to baseline situation (p < 0.0001). The gel was rated best, the carmellose spray was rated worst by the patients, but the single compounds did not differ significantly in their effects. In spite of this result, most patients chose the carmellose spray as their favorite compound. This is due to its good taste and easy handling, which play an important role for the acceptance of the products. Big individual differences in the preference of the single compounds were found. CONCLUSION: For most patients considerable relief from xerostomia can be reached by saliva substitutes. Thus, every patient with xerostomia should be given different artificial saliva compounds for a test period. This will help to find the individually best way to cope with the dry mouth. PMID- 15827693 TI - Amifostine is a potent radioprotector of salivary glands in radioiodine therapy. Structural and ultrastructural findings. AB - BACKGROUND AND PURPOSE: Salivary gland impairment following high-dose radioiodine treatment is well recognized. Since differentiated thyroid cancer has a good prognosis, reduction of long-term side effects is important. This study investigates the radioprotective effects of amifostine in salivary glands of rabbits receiving high-dose radioiodine therapy so as to obtain deeper insight in changes on the cellular and ultrastructural level. MATERIAL AND METHODS: A total of 20 rabbits were investigated. High-dose radioiodine therapy applying 1 GBq (131)I was performed in 16 animals. Eight of these 16 animals received 200 mg/m(2) body surface amifostine prior to high-dose radioiodine therapy. Four additional rabbits served as controls, two receiving amifostine, the other two no treatment at all. Subsequently, salivary glands (submandibular and parotid glands, respectively) of all animals were examined by light and transmission electron microscopy. RESULTS: Parenchymal damage of both parotid and submandibular glands, specially acinar structures comprising cell organelles and nuclei, of animals pretreated with amifostine was much less pronounced than in animals without amifostine pretreatment. CONCLUSION: The results indicate that parenchymal damage in salivary glands induced by high-dose radioiodine therapy can significantly be reduced by amifostine. Therefore, amifostine therapy may increase quality of life in patients with differentiated thyroid cancer after radioiodine treatment. PMID- 15827694 TI - Postoperative irradiation of incompletely excised gemistocytic astrocytomas. Clinical outcome and prognostic factors. AB - BACKGROUND AND PURPOSE: Although gemistocytic astrocytomas are considered slow growing tumors, they often behave aggressively and carry the least favorable prognosis among low-grade astrocytomas. The aim of this study is to evaluate the outcomes and prognostic factors of patients with incompletely excised gemistocytic astrocytomas irradiated postoperatively. PATIENTS AND METHODS: Records of 48 patients with incompletely excised gemistocytic astrocytoma, irradiated between 1976 and 1998 at the Department of Radiation Oncology, Maria Sklodowska-Curie Memorial Cancer Center, Cracow, Poland, were reviewed. The total dose ranged from 50 to 60 Gy (mean: 59.35, median: 60 Gy) delivered in daily fractions of 2 Gy, 5 days a week. The treatment volume covered the residual tumor with a margin of 1-2 cm. RESULTS: Toxicity was acceptable. The overall actuarial survival rates at 5 and 10 years were 30% and 17%, respectively. Age and gender had an influence on overall survival by univariate and multivariate analysis (p < 0.05). Patients < or = 35 years of age and female patients carried the best prognosis. CONCLUSION: In most patients with gemistocytic astrocytoma, combined surgery and postoperative radiotherapy result in only short-term survival. Older age is the most important unfavorable prognostic factor in patients with gemistocytic astrocytoma. PMID- 15827695 TI - Is a diagnostic CT of the brain indicated in patients with choroidal metastases before radiotherapy? AB - BACKGROUND AND PURPOSE: There is no evidence in the literature about the incidence of synchronous brain metastases in patients with choroidal metastases. This is of major importance, because the radiation fields of choroidal metastases and, later on, brain metastases, if treated consecutively, are partly overlapping, thus potentially increasing the rate of late side effects such as brain necrosis. The goal of this study was to determine the frequency of synchronous brain metastases. PATIENTS AND METHODS: 50 patients with choroidal metastases were enrolled into a study of the "Arbeitsgemeinschaft Radiologische Onkologie" of the German Cancer Society (ARO 95-08) with standardized 40 Gy radiotherapy, 2 Gy single dose. All patients were staged before treatment with a computed tomography of the brain (CCT). No patient showed clinical signs of brain metastases. RESULTS: 13 out of 50 patients (26%) had brain metastases in the CCT leading to radiotherapy of the brain and choroidal metastases in one volume. CONCLUSION: A CCT is indicated at the diagnosis of choroidal metastases for screening of synchronous brain metastases. The incidence is about 25%, and the diagnosis of brain metastases results in a different target volume: the whole brain including the posterior parts of the eyes compared to the posterior parts of the eyes alone. Therefore, the risk of late side effects could be reduced compared with an additional later radiotherapy of the whole brain with partly overlapping fields. PMID- 15827696 TI - [Volumetric changes of the breast during radiotherapy. Is a replanning necessary for the electron boost?]. AB - BACKGROUND AND PURPOSE: Radiotherapy can induce tissue reactions with an edema leading to increased breast volume. The aim of the present study was to quantify this increase and analyze its effect on the electron boost technique. PATIENTS AND METHODS: 140 patients with breast cancer treated with breast-conserving surgery underwent CT planning before, during and/or after radiotherapy in order to evaluate breast volume changes due to radiotherapy. CT data were analyzed using the HELAX planning system and dose distribution was assessed. Determination of the breast volume was achieved using an interpolation algorithm. Three subgroups were analyzed: group 1 (n = 47): < or = 670 cm(3), group 2 (n = 46): 671-999 cm(3), and group 3 (n = 47): > or = 1000 cm(3) breast volume. RESULTS: The mean initial breast volume was 907 cm(3) (100-3073 cm(3)). After radiotherapy, mean breast volume increased by 81 cm(3) to 988 cm(3) (109-3185 cm(3)). Significant changes in volume were observed after a dose of 40 Gy. According to the subgroups mean volume increase was as follows: group 1: 53 cm(3) (3-120 cm(3)), group 2: 85 cm(3) (20-200 cm(3)), and group 3: 105 cm(3) (5-340 cm(3)). This difference was statistically significant for all subgroups (p < 0.001). Corresponding to the volume increase, depth of the boost target volume changed up to 1.0 cm. CONCLUSION: As radiotherapy may lead to a significant increase in breast volume, it seems appropriate to perform a second planning CT after about 40 Gy in order to optimize dose distribution for boost irradiation. PMID- 15827697 TI - Radiotherapy and chemotherapy for myoepithelioma of the sellar region. AB - BACKGROUND: Myoepithelioma might arise in the head and neck area, especially within the salivary glands. It is very uncommon as a primary intracranial tumor. CASE REPORT: A 34-year-old African patient with proliferating myoepithelioma originating in the sellar region is described. After subtotal resection, the tumor recurred locally despite postoperative radiotherapy to a total dose of 54 Gy (Figures 1 and 2). When intracranial metastases developed, chemotherapy with ifosfamide and, later, BCNU was administered. No objective response was obtained with any of the nonsurgical approaches. Uncontrolled intracranial tumor growth led to the patient's death 20 months after the initial diagnosis. CONCLUSION: Most tumors of the sellar region have a favorable prognosis. However, this case of incompletely resected proliferating myoepithelioma showed both local and distant recurrences, which did not respond to further treatment. Thus, complete surgical resection is recommended, whenever technically feasible. PMID- 15827698 TI - Evaluation of transversal slice imaging in the diagnosis of tooth displacement with special consideration of the upper canines. AB - AIM: The exact localization and analysis of displaced, retained or impacted teeth can rarely be achieved by common radiologic methods providing eccentric or occlusal images. The aim of this study was to evaluate transversal slice imaging (TSI) as a diagnostic alternative, keeping radiation exposure under consideration. MATERIAL AND METHODS: In a pilot study, we simulated seven different canine positions in a phantom head. Each tooth position was visualized by transversal slice imaging and compared by several observers independently and blinded to computer tomography (CT) serving as the gold standard. The clinical study included the retrospective analysis of TSI samples from 27 patients (mean age 13.7 yrs.) according to these criteria: 1. location of the canine crown and root, 2. recognition of the crown, 3. stage of root development, 4. root dilaceration, 5. periodontal ligament space, 6. ankylosis, 7. relationship to neighboring teeth, and 8. resorptions of neighboring teeth. The evaluations of canine position were compared to the results gained intraoperatively. RESULTS: In the transversal slice images taken from a phantom head, the observers' estimations of the canine position were identical in five cases (71.4%). At least four observers using TSI described identical tooth positions in 89% of patients. In 2.5%, no statement concerning tooth position was feasible. Crown recognition was possible in 75% of cases. Ankylosis, periodontal ligament space, root development, dilacerations and resorptions were not assessable in the majority of the cases. CONCLUSION: Transversal slice imaging can be recommended as a second plane projection in diagnosing tooth displacement, especially concerning oro vestibular localization (effective radiation dose 0.003-0.012 mSv). However, as far as periodontal ligament space, resorptions and root morphology are concerned, this method has its limits. PMID- 15827699 TI - Three-dimensional soft tissue prediction using finite elements. Part I: Implementation of a new procedure. AB - BACKGROUND AND AIM: The prediction of soft tissue esthetics is important for achieving an optimal esthetic outcome in orthodontic treatment planning. Applicable procedures have so far been restricted to two-dimensional profile predictions that have not proven to be very reliable. The goal of this investigation was therefore to develop a novel finite element-based procedure that allows a three-dimensional, easily visualized, quantitative analysis and prediction of soft tissue behavior for the clinician. The procedure to be developed should be easy to handle and not entail any additional radiation exposure for the patient. MATERIAL AND METHODS: Using a three-dimensional scanner, the facial surfaces of 20 probands were digitalized and individual FEM models were generated. RESULTS: After reduction of data redundancy via several conversion steps, a patient-specific simulation model was prepared consisting of 20,000 to 40,000 individual elements to which specific physical properties could be assigned. The average time required for generating a virtual model was 50 minutes. Problems occurring during model generation were rare (mainly shadowing phenomena and movement artifacts). CONCLUSION: The procedure outlined herein makes the reliable generation of patient-specific simulation models possible for facial soft tissue prediction in orthodontics. PMID- 15827700 TI - Three-dimensional soft tissue prediction using finite elements. Part II: Clinical application. AB - BACKGROUND AND AIM: The goal of this study was to analyze the validity and prediction accuracy of a newly-developed procedure for three-dimensional soft tissue prediction based on Finite Element Method, and to compare the results with prediction produced using an existing two-dimensional prediction program (Dentofacial Planner Plus). PATIENTS AND METHODS: In twelve patients who underwent combined surgical-orthodontic treatment, profile prediction was generated using both procedures preoperatively and then compared at predefined measurement points with the patient's actual postoperative soft tissue status. RESULTS: The deviations observed depended on the facial region, whereby the prediction errors for both procedures were much greater in the lower facial third than in the midfacial third. Calculating in all the measurement points, the mean horizontal prediction error was 0.32 mm for the Finite Element Method and 0.75 mm for the Dentofacial Planner Plus. Overall, we were able to demonstrate the new procedure's superior validity and quality of visualization. In addition to profile prediction, the procedure allows a differentiated three-dimensional assessment of esthetically important regions such as the cheeks, nasolabial folds and the nasal wings. Additional X-radiation is not necessary in this risk-free and stress-free procedure. CONCLUSION: Three-dimensional soft tissue prediction employing finite element modeling is a useful aid for implementing esthetically optimized treatment planning. PMID- 15827701 TI - Dental arch and cephalometric changes following treatment for class III malocclusion by means of the function regulator (FR-3) appliance. AB - AIM: The aim of this retrospective clinical study was to evaluate dental arch, skeletal, dentoalveolar, and soft tissue profile changes following treatment of Class III malocclusion by means of the Function Regulator (FR-3) appliance. SUBJECTS AND METHODS: The study sample consisted of 14 growing patients (seven boys and seven girls) with a mean age of 9.7 +/- 1.1 years presenting Class III malocclusion. They were treated with an FR-3 appliance exclusively for a period of 2.4 +/- 0.6 years. Dental casts and lateral cephalometric radiographs were obtained immediately before initiation and after completion of treatment with the FR-3 appliance. Thirteen variables were used for the dental cast analysis and 61 variables for the cephalometric analysis. The paired t-test was utilized to evaluate the presence of significant changes (p < 0.05). RESULTS: The evaluation of the dental casts revealed that there was a significant increase in intermolar, interpremolar, and intercanine width of the maxilla and of palatal height after treatment. Concerning the mandible, an increase in intermolar and intercanine width and a decrease in lower arch depth were observed. Cephalometric evaluation revealed a significant decrease in SNB angle and an increase in ANB angle, overjet, facial convexity, nose prominence, and lower soft tissue face height. There was an increase in upper lip thickness and a decrease in lower lip convexity observed after treatment. CONCLUSIONS: The results of this study confirm a favorable functional and esthetic maxillary and mandibular position following treatment by means of the function regulator (FR-3) appliance. PMID- 15827702 TI - Frequency of and complications in the use of RPE appliances--results of a survey in the Federal State of Hesse, Germany. AB - AIM: A questionnaire was sent to orthodontists with private practices in the German State of Hesse to obtain information about the frequency of rapid palatal expansion and complications during treatment with rapid palatal expansion appliances (RPE-A). MATERIAL AND METHOD: A series of questions therein inquired as to the types of RPE-As used, their activation rhythm, the extent and the duration of the active expansion and retention; what their age limit was, starting from when surgical assistance was deemed necessary. Another series of queries concerned any technical and medical complications observed. RESULTS: The most frequently used type was the Hyrax screw (75%) soldered to bands. Technical problems arose with all applied indicated types. Although the answers to the first series of questions showed great variance, the majority of the orthodontists activated the screw twice a day (range 1-6 times a day) by a quarter turn for 2-3 (range 1-12) weeks. The retention phase amounted to 12 (range 4-26) weeks and the age limit for the indication of a surgical intervention was on average 17 years, with a range of 10-35 years. Medical complications such as pain and decubitae were reported by a third of the inquired offices, a non-opening suture was observed in one of every five practices. This meant that 1.7% of the sutures failed to open, and tooth tilting and rarely, extreme root resorptions and/or bone dehiscences were observed. Solid carious lesions were observed in one case after removal of an adhesive RPE. Extraoral changes such as those in the nose were reported by one in every ten practices. CONCLUSIONS: The RPE represents a common treatment means during which side effects are often temporary and permanent damages rather rare. It is important to inform the patient about both positive effects and possible complications, in particular pain and extraoral changes. PMID- 15827703 TI - Motivation, acceptance and problems of invisalign patients. AB - AIM: This study examined how well patients accepted and to what extent they were impaired by Invisalign treatment. PATIENTS AND METHODS: Fifty-four consecutive patients received a questionnaire with 12 questions after 3 to 6 months of Invisalign treatment. They were given a choice of three responses for each question. Apart from personal data, the questionnaire covered the following parameters: adaptation time, occurrence and duration of pain, possible speech impairment, lingual and mucosal irritations, temporomandibular joint (TMJ) problems and subjective assessment of the success of therapy thus far and the quality of information initially provided. RESULTS: 78% of those interviewed were females. The highest percentage of patients (44%) were between 20 and 30 years of age. 83% got used to their aligners within one week. 35% had no pain and 54% mild pain while wearing them. This pain usually lasted for 2 to 3 days following insertion of a new aligner. 46% of the patients experienced no speech impairment. 93% felt so secure with their aligners that they felt not at all inhibited about speaking. The majority noticed no narrowing of the lingual space (76%) or irritation of the lingual or buccal mucosa (70%). However, 6% had strong irritations. 44% of the patients had difficulty chewing, mainly because the teeth were sensitive to pressure or had food particles caught between them due to temporary gaps. TMJ problems in terms of clicking were reported by 8% of the patients, although the clicking had existed before therapy initiation. None of the patients had TMJ pain. At the time of the interview, 89% of the patients were satisfied with the progress of therapy. All patients considered themselves well or very well informed about the treatment. CONCLUSIONS: Invisalign therapy seems particularly attractive to a clientele comparable to that for the lingual technique. Patients show high acceptance, since they become accustomed to the aligners very quickly and do not suffer much impairment. If indicated, Invisalign therapy should thus be considered an alternative to the lingual technique for esthetically-demanding patients. This applies especially to patients who cannot be treated by the lingual technique because of the inevitable speech impairment. PMID- 15827704 TI - [Cartilage cell transplantation as an alternative to endoprosthesis]. AB - The incidence of cartilage lesions is high. Due to pain and loss of function, long-term therapy is often necessary. Isolated, full-thickness articular cartilage lesions with a diameter of 4 cm2 are indications for chondrocyte transplantation. Many outcome studies report good long-term results. In recent years, the cultivation of chondrocytes has changed. Histologic investigations show hyalin-like cartilage after transplantation. Large cartilage lesions facing each other are a therapeutic dilemma in young patients. Implanting unicondylar or bicondylar prostheses is sometimes the last resort for these patients, and there are several reports of good clinical outcome. Cartilage transplantation has until now been recommended for patients younger than 50. It is not an alternative to joint replacement. PMID- 15827706 TI - [Tracheotomy and tracheostomy techniques]. AB - Tracheostomy has become the method of choice in managing patients requiring long term mechanical ventilation. At present, there are several alternatives to conventional surgical tracheostomy such as percutaneous dilatational techniques according to Ciaglia, Frova, and Fantoni. The basic principle of these new techniques is percutaneous puncture of the trachea and subsequent dilatation of the puncture channel until the tracheal cannula can be inserted. The advantages are "bedside" performance in the intensive care unit and the use of minimal technical equipment. Nevertheless, dilatation tracheostomy is not always technically feasible and safe. Some significant complications and contraindications have been recognized recently. This should be taken into account when planning tracheostomy in long-term ventilated patients. Compared to conventional surgical tracheostomy, this new procedure retains its value and benefit only if these contraindications are carefully observed. PMID- 15827705 TI - [Role of stem cells in adult hepatic regeneration]. AB - Central to the successful surgically treatment numerous liver diseases is the ability of the organ to regenerate. The understanding of the process of self renewal has both changed and progressed over the last few decades. For many years, the assumption was that the liver regenerates primarily through the division of mature liver cells. However, over the last few years there has been increasing evidence of the participation of stem cells. Intrahepatic stem cells, so-called oval cells, are activated under conditions of severe or chronic liver disease and originate from the canals of Hering. In addition, extrahepatic stem cells may migrate from the bone marrow into the liver when the regenerative capacity of the liver itself is depleted. It is not yet fully clear how the different stem cell populations interact with both each other and the mature liver cell population to achieve homeostatic cell and differentiation equilibrium in the diseased and/or regenerating organ. In any case, the outstanding growth potential of liver stem cells may become a clinically viable option in the field of cell transplantation. PMID- 15827707 TI - [Lymph vascularity and lymph node metastases on PET and PET-CT: immunohistological and clinical observations]. AB - Sarcomatous malignancies only rarely develop regional lymph node metastases: about 2.7% of our evaluated cases. In this paper we provide evidence supporting a new hypothesis that two entirely separate lymph vascular systems exist in humans. One system (LGS I) exists in close proximity to the epithelium and drains into regional lymph nodes. Only sarcomas that originate in the epithelium or its immediate proximity are able to form regional lymph node metastases. The vast majority of sarcomatous malignancies (97.4% of cases) do not give rise to lymph node metastases, since they originate in proximity to a second, more deeply localized lymph node system (LGS II) in the mesenchymally derived tissues of the body. This second system has no connection to regional lymph nodes. Supporting evidence is provided by experience in the operative treatment of extremity lymphedema, PET-CT examinations, radionuclear lymphography, and scientific investigations using antibodies specifically directed at the elements of the lymph vascular system. PMID- 15827708 TI - [Stenting in acute stroke]. AB - Hemodynamic impairment caused by pseudo-occlusion or occlusion of the internal carotid artery (ICA) is a rare etiology of stroke. Acute revascularization of the vessel can restore the flow to the brain. This retrospective study shows that ICA stenting with proximal and distal protection devices in patients with acute stroke and pseudo-occlusion or occlusion of the ICA is technically feasible and can prevent further hemodynamic strokes. In contrast, selected patients have a high risk of reperfusion syndrome with intracranial hemorrhage and peri interventional monitoring of blood pressure in an ICU is recommended. PMID- 15827709 TI - [Integrated care. Taking a stand]. PMID- 15827710 TI - [The Association of Statutory Health Insurance Physicians in 10 years. Where does the path of medical and psychotherapeutic self administration lead?]. PMID- 15827711 TI - [Implantation of the AMS 800 artificial sphincter system in a woman via a paraurethral access]. AB - The procedure described demonstrates the feasibility of reliable implantation of an artificial sphincter system in female patients via a paraurethral access between the urethral orifice and the clitoris following radical cystectomy with continent, orthotopic urinary diversion and postoperatively developed urinary stress-incontinence. The technique presented avoids the more invasive transperitoneal or transvaginal approaches which jeopardize the neobladder-neck and increase the risk for infections or fistulas. PMID- 15827712 TI - [Perspectives in public health. The future of the specialist]. PMID- 15827716 TI - Extraction procedures for chemical speciation of arsenic in atmospheric total suspended particles. AB - An arsenic chemical speciation study was performed in 2000, using air filters on which total suspended particles (TSP) were collected, from the city of Huelva, a medium size city with huge industrial influence in SW Spain. Different procedures for extraction of the arsenic species were performed using water, NH2OH.HCl, and H3PO4 solutions, with either microwave or ultrasonic radiation. The best optimised extraction methods were use of 100 mmol L(-1) NH2OH.HCl and 10 mmol L( 1) H3PO4 and microwave radiation for 4 min. High-performance liquid chromatography coupled with hydride generation and atomic fluorescence spectrometry (HPLC-HG-AFS) was employed for determination of the arsenic species. The results from 12 TSP air filters collected on a monthly basis showed extraction was quantitative (94% with NH2OH.HCl and 86% H3PO4). Only inorganic arsenic species (arsenite and arsenate) were detected. The mean arsenite concentration was 1.2+/-0.3 ng m(-3) (minimum 0.3 ng m(-3), maximum 1.8 ng m( 3)). The mean arsenate concentration was 10.4+/-1.8 ng m(-3), with greater monthly variations than arsenite (minimum 2.1 ng m(-3), maximum 30.6 ng m(-3)). The high level of arsenic species in the TSP samples can be related to a copper smelter located in the region. PMID- 15827719 TI - Flow-injection technique for determination of uranium and thorium isotopes in urine by inductively coupled plasma mass spectrometry. AB - A sensitive and efficient flow-injection (FI) preconcentration and matrix separation technique coupled to sector field ICP-mass spectrometry (SF-ICP-MS) has been developed and validated for simultaneous determination of ultra-low levels of uranium (U) and thorium (Th) in human urine. The method is based on selective retention of U and Th from a urine matrix, after microwave digestion, on an extraction chromatographic TRU resin, as an alternative to U/TEVA resin, and their subsequent elution with ammonium oxalate. Using a 10 mL sample, the limits of detection achieved for 238U and 232Th were 0.02 and 0.03 ng L(-1), respectively. The accuracy of the method was checked by spike-recovery measurements. Levels of U and Th in human urine were found to be in the ranges 1.86-5.50 and 0.176-2.35 ng L(-1), respectively, well in agreement with levels considered normal for non-occupationally exposed persons. The precision obtained for five replicate measurements of a urine sample was 2 and 3% for U and Th, respectively. The method also enables on-line measurements of the 235U/238U isotope ratios in urine. Precision of 0.82-1.04% (RSD) was obtained for 235U/238U at low ng L(-1) levels, using the FI transient signal approach. PMID- 15827720 TI - Evaluation of a microwave-assisted extraction technique for determination of water-soluble inorganic species in urban airborne particulate matter. AB - A simple and rapid microwave-assisted extraction (MAE) technique has been developed for the determination of water-soluble inorganic species (cations: Na+, NH4+, K+, Ca2+ and Mg2+ and anions: F-, Cl-, NO3-, PO4(3-) and SO4(2-)) in airborne particulate matter. The analytes were extracted under different treatment conditions such as microwave power and extraction time. They were quantified using ion chromatography. The observed concentrations and recovery yields obtained under different conditions were compared. The results of a comparison between this MAE and sonication using NIST SRM 1648 are also given in this paper. The optimized MAE technique gave results in good agreement with the values obtained by the sonication. For some ions, for example Mg2+ and K+, recovery was low with both techniques. The results demonstrated that the optimized MAE is fast and efficient compared with conventional ultrasonic extraction. Urban airborne particles were collected and subjected to the MAE followed by the IC analysis to determine the relative proportions of different water-soluble inorganic species. These results are briefly discussed. PMID- 15827721 TI - Validation of a screening method for the simultaneous identification of fat soluble and water-soluble vitamins (A, E, B1, B2 and B6) in an aqueous micellar medium of hexadecyltrimethylammonium chloride. AB - Simultaneous determination of the fat-soluble vitamins A and E and the water soluble vitamins B1, B2 and B6 has been carried using a screening method from fluorescence contour graphs. These graphs show different colour zones in relation to the fluorescence intensity measured for the pair of excitation/emission wavelengths. The identification of the corresponding excitation/emission wavelength zones allows the detection of different vitamins in an aqueous medium regardless of the fat or water solubility of each vitamin, owing to the presence of a surfactant which forms micelles in water at the used concentration (over the critical micelle concentration). The micelles dissolve very water insoluble compounds, such as fat-soluble vitamins, inside the aggregates. This approach avoids the use of organic solvents in determining these vitamins and offers the possibility of analysing fat- and water-soluble vitamins simultaneously. The method has been validated in terms of detection limit, cut-off limit, sensitivity, number of false positives, number of false negatives and uncertainty range. The detection limit is about microg L(-1). The screening method was applied to different samples such as pharmaceuticals, juices and isotonic drinks. PMID- 15827722 TI - Electrochemical determination of the total antioxidant capacity of human plasma. AB - Electrochemical reduction of oxygen at a glassy carbon electrode in a 0.05 mol L( 1) solution of (C2H5)4NI in dimethylformamide leads to generation of the superoxide anion-radical. This product of reversible one-electron oxygen reduction reacts with antioxidants, a process which is based on protonation of the anion-radical by the antioxidant. Rate constants of this interaction have been calculated. Human plasma antioxidants also react with electrochemically generated superoxide anion-radical. A voltammetric method is proposed for estimation of the total antioxidant capacity (TAC) of plasma on the basis of on this reaction. The TAC of plasma was also determined using constant-current coulometry with electro-generated bromine as the active species. A correlation was observed between TAC data obtained by voltammetry (O2*-, in alpha-tocopherol units) and coulometry (Br2 as titrant). TAC of plasma from patients with purulent infections was determined. Statistically significant differences were found between TAC of patients and control group. Treatment of purulent infections increases the TAC of plasma. So, use of electrochemical methods (voltammetry and coulometry) for determination of TAC can be used for estimation of the effectiveness of treatment. PMID- 15827723 TI - Order and disorder in alkyl stationary phases. AB - Covalently modified surfaces represent a unique state of matter that is not well described by liquid or solid phase models. The chemical bond in tethered alkanes imparts order to the surface in the form of anisotropic properties that are evident in chromatographic and spectroscopic studies. An understanding of the structure, conformation, and organization of alkyl-modified surfaces is requisite to the design of improved materials and the optimal utilization of existing materials. In recent years, the study of alkyl-modified surfaces has benefited from advances in modern analytical instrumentation. Aspects of alkyl chain conformation and motion have been investigated through the use of nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, Raman spectroscopy, fluorescence spectroscopy, and neutron scattering studies. Chromatography provides complementary evidence of alkyl chain organization through interactions with solute probes. Computational simulations offer insights into the structure of covalently modified surfaces that may not be apparent through empirical observation. This manuscript reviews progress achieved in the study of the architecture of alkyl-modified surfaces. PMID- 15827724 TI - Electrochemical characterisation of dental alloys: its possibilities and limitations. AB - Dental alloys are metallic biomaterials which have a broad variation of composition compared to technical alloys. It is therefore in the interest of patients and technicians to conduct a good assessment of the electrochemical behaviour of dental alloys in order to collect information about their corrosion resistance. The purpose of this work was to demonstrate possibilities and limitations of two electrochemical techniques: the voltammetry of immobilised microparticles (ViMP) onto lead, and cyclic voltammetry measurements with the help of the mini-cell system (MCS). Based on fingerprints obtained from ViMP it was possible to analyse and differentiate the dental alloys. The results obtained by MCS were comparable with ViMP, but give a better understanding of the corrosion behaviour of the materials. PMID- 15827725 TI - Determination of sterols in biological samples by SPME with on-fiber derivatization and GC/FID. AB - A new procedure for the determination of sterols in serum samples is proposed. The system consists of coating a Solid Phase Microextraction (SPME) microfiber in headspace mode with the derivatizing agent N,O bis(trimethylsilyl)trifluoracetamide (BSTFA) and then applying this coated fiber to the simultaneous extraction and derivatization of three precursors in the cholesterol biosynthesis pathway (desmosterol, lathosterol and lanosterol) and two phytosterols (sitosterol and sitostanol) in serum samples. Optimization of the analytical procedure via the application of an experimental design, a study of matrix effects, and an analysis of serum pool samples are all described and discussed. PMID- 15827726 TI - Delayed development of silicoproteinosis with diffuse interstitial fibrosis: 16 year follow-up with autopsy correlation. PMID- 15827727 TI - Radiofrequency ablation in the liver using two cooled-wet electrodes in the bipolar mode. AB - The purpose of this study was to demonstrate the efficacy of bipolar radiofrequency ablation (RFA) using cooled-wet electrodes inducing coagulation in ex vivo bovine livers and in in vivo canine livers. In ex vivo experiments, 20 coagulations were created by monopolar (group A), and bipolar RFA (group B) using a 200 W generator (Valleylab) and one or two cooled-wet electrodes. In in vivo experiments, one coagulation was created by bipolar RFA in each of eight dogs via laparotomy. In ex vivo and in vivo experiments, RF was applied to one or two electrodes at 100 W for 10 min. The dimensions of the coagulations were compared in the two groups. In ex vivo experiments, the mean volumes of the coagulations produced in group B (54.0+/-16.5 cm3) were greater than those produced in group A (33.9+/-12.7 cm3) (P=0.007). In in vivo experiments, bipolar RFA produced a coagulation of 39.4+/-15.6 cm3 without a major complication. The present study showed that a RF electrode system using two cooled-wet electrodes in the bipolar mode created larger coagulation volumes than the monopolar mode, and this system can be used to create large coagulation without major complications. PMID- 15827728 TI - A pilot study of bone density loss in menopausal women treated with chemotherapy for cancer. AB - GOALS OF WORK: To estimate the incidence and severity of bone loss in menopausal women diagnosed with cancer who receive treatment with chemotherapy. Also, to evaluate the use of bone loss prevention agents in this population. PATIENTS AND METHODS: A total of 25 postmenopausal women with newly diagnosed cancers who received chemotherapy for a minimum of six cycles were enrolled in this pilot study. All subjects underwent baseline bone mineral density (BMD) testing of the lumber spine (LS), left hip (LH), and femoral neck (FN). Of the 25 women, 22 also underwent follow-up BMD testing at 6 months. MAIN RESULTS: The median age of the subjects was 61 years (range 41-76 years) and the median age of menopause was 50 years (range 34-55 years). Of the 25 subjects, 9 used at least 1 g oral calcium daily, 4 used alendronate, 2 used raloxifene, and 1 used oral estrogen. The mean BMDs (g/cm2) with standard deviation above or below the mean for young adult women at baseline were: LS 0.996 (-0.5 SD), LH 0.876 (-0.5 SD), and FN 0.760 ( 0.7 SD). The following values were obtained at 6 months: LS 0.965 (P<0.001), LH 0.847 (P<0.001), and FN 0.739 (P=0.009). CONCLUSIONS: Menopausal women diagnosed with cancer appear to have a high incidence of baseline bone loss, with significant additional loss during treatment. Use of agents for prevention/treatment of bone loss in this group is infrequent. A prospective, controlled analysis is indicated to determine the optimal utility of bone densitometry testing and osteoporosis prevention strategies in this population. PMID- 15827729 TI - Nurses' willingness to maximize opioid analgesia for severe cancer pain, and its predictor. AB - GOALS OF WORK: The effectiveness of cancer pain management (CPM) is influenced by nurses' willingness to maximize opioid analgesia for severe cancer pain. The purposes of this study were to identify the willingness of nurses to provide maximum-dose opioids whenever needed for CPM and to determine its associated predictors. METHODS: This multicenter study was conducted among the entire total of registered nurses in seven large hospitals in Korea. Its overall response rate was 41.6%, and the data from 930 who responded (40.1%) were analyzed. We utilized a three-step, multidimensional, multiple logistic regression to identify the predictors of nurses' willingness. MAIN RESULTS: Only 255 nurses (27.4%) indicated that they recommended the maximum dose of opioids whenever it was needed. The respondents who were more likely to recommend morphine showed the following characteristics: older nurses (odds ratio, OR, 1.57; confidence interval, CI, 1.13-2.19); they knew the effectiveness of opioids for CPM (OR 1.53; CI 1.06-2.20); rarely concerned about a patient's addiction to opioids (OR 2.16; CI 1.48-3.15), or to a family member's addiction (OR 1.81; CI 1.20-2.73); prior experience with pain assessment tools (OR 1.62; CI 1.11-2.37); practical experience caring for cancer patients with pain over 51% (OR 1.55; CI 1.09-2.19). CONCLUSIONS: Our multicenter study suggested that in order to improve nurses' willingness to recommend opioids liberally in CPM: (1) attitudes about fear of opioid addiction must be changed; (2) the efficiency of opioids in CPM must be taught; and (3) implementation of pain assessment tools must be undertaken. PMID- 15827731 TI - Effect of sulfite on red blood cell deformability ex vivo and in normal and sulfite oxidase-deficient rats in vivo. AB - The effect of sulfite, a widely used food additive, on red blood cell deformability ex vivo and in vivo was investigated. Ex vivo experiments were conducted in human blood exposed to sulfite (5, 10 and 20 mM). In vivo experiments were carried out in sulfite oxidase-competent (SOXC) and sulfite oxidase-deficient (SOXD) rats. In the in vivo experiments, sulfite was administered in the form of sodium metabisulfite (Na(2)S(2)O(5), 25 mg/kg/day) via drinking water. Vitamin E dissolved in olive oil at a dose of 50 mg/kg was administered by gastric gavages. Red blood cell (RBC) deformability was determined at various fluid shear stresses using an ektacytometer. Ex vivo sulfite exposure to RBC did not affect RBC deformability. In the in vivo experiments, although RBC deformability was not affected by sulfite treatment in SOXD rats, it was found to be significantly increased in SOXC rats. Vitamin E treatment in combination with sulfite caused impairment in RBC deformability in both SOXC and SOXD rats. We suggest that sulfite needs to be oxidized in order to improve RBC deformability. PMID- 15827730 TI - Oxoiron(IV) porphyrin pi-cation radical complexes with a chameleon behavior in cytochrome P450 model reactions. AB - There is an intriguing, current controversy on the involvement of multiple oxidizing species in oxygen transfer reactions by cytochromes P450 and iron porphyrin complexes. The primary evidence for the "multiple oxidants" theory was that products and/or product distributions obtained in the catalytic oxygenations were different depending on reaction conditions such as catalysts, oxidants, and solvents. In the present work, we carried out detailed mechanistic studies on competitive olefin epoxidation, alkane hydroxylation, and C=C epoxidation versus allylic C-H hydroxylation in olefin oxygenation with in situ generated oxoiron(IV) porphyrin pi-cation radicals (1) under various reaction conditions. We found that the products and product distributions were markedly different depending on the reaction conditions. For example, 1 bearing different axial ligands showed different product selectivities in competitive epoxidations of cis olefins and trans-olefins and of styrene and para-substituted styrenes. The hydroxylation of ethylbenzene by 1 afforded different products, such as 1 phenylethanol and ethylbenzoquinone, depending on the axial ligands of 1 and substrates. Moreover, the regioselectivity of C=C epoxidation versus C-H hydroxylation in the oxygenation of cyclohexene by 1 changed dramatically depending on the reaction temperatures, the electronic nature of the iron porphyrins, and substrates. These results demonstrate that 1 can exhibit diverse reactivity patterns under different reaction conditions, leading us to propose that the different products and/or product distributions observed in the catalytic oxygenation reactions by iron porphyrin models might not arise from the involvement of multiple oxidizing species but from 1 under different circumstances. This study provides strong evidence that 1 can behave like a "chameleon oxidant" that changes its reactivity and selectivity under the influence of environmental changes. PMID- 15827733 TI - Increase in reaction time for the peripheral visual field during exercise above the ventilatory threshold. AB - The purpose of the present study was to determine whether reaction time (RT) for the peripheral visual field increases at exercise intensity above the ventilatory threshold (VT) during incremental exercise and to examine the relationship between aerobic capacity and the extent of increase in the RT. Nine healthy subjects performed a simple manual RT task for the peripheral visual field at rest, during exercise on a cycle ergometer, and immediately after exercise. After warm-up exercise, the subjects cycled at 40 W for 3 min, increasing by 40 W every 3 min until 240 W in a step-wise manner. During incremental exercise, RT measurements were performed 1 min and 30 s after the start of every increase in workload. The RT for the peripheral visual field significantly increased at exercise intensity above VT, as compared with at rest. The increase in the RT, which was calculated by subtracting the RT at rest from the RT at 240 W, negatively correlated with maximal oxygen uptake VO(2max) for each subject (r = 0.73, P < 0.05). It is likely that high aerobic capacity attenuates the increase in the RT for the peripheral visual field during exhaustive exercise. PMID- 15827734 TI - Coordination of head and trunk accelerations during walking. AB - The purpose of this study was to investigate the relationship between oscillatory dynamics of the head and trunk in each plane of motion during walking. Head and trunk accelerations of ten healthy subjects (age: 23 +/- 4 years) were measured in the vertical (VT), anterior-posterior (AP) and mediolateral (ML) directions using a pair of tri-axial accelerometers. All subjects performed five walking trials along a level 20 m walkway at their preferred gait velocity (1.30 +/- 0.15 m s(-1)). Acceleration data were analysed using power spectral, harmonic and regularity measures. Results indicated that: (1) oscillations of the head were smoother, with a greater proportion of power at lower frequencies than oscillations of the trunk, (2) differences in power spectral properties between the head and trunk were most pronounced in the ML direction, (3) coupling between VT-AP, VT-ML, and AP-ML accelerations were greater for the head than trunk, and (4) for both segments, the weakest coupling was observed for AP-ML acceleration relations. Overall, the results of this study suggest that accelerations of the head are significantly attenuated, and more tightly controlled, compared to accelerations of the lower trunk. This attenuation process was particularly evident for the ML direction, whereby head accelerations showed the greatest differences compared to ML accelerations at the trunk. PMID- 15827735 TI - Reliability of peak VO(2) and maximal cardiac output assessed using thoracic bioimpedance in children. AB - The purpose of this study was to evaluate the reliability of a thoracic electrical bioimpedance based device (PhysioFlow) for the determination of cardiac output and stroke volume during exercise at peak oxygen uptake (peak VO(2) in children. The reliability of peak VO(2) is also reported. Eleven boys and nine girls aged 10-11 years completed a cycle ergometer test to voluntary exhaustion on three occasions each 1 week apart. Peak VO(2) was determined and cardiac output and stroke volume at peak VO(2) were measured using a thoracic bioelectrical impedance device (PhysioFlow). The reliability of peak VO(2) cardiac output and stroke volume were determined initially from pairwise comparisons and subsequently across all three trials analysed together through calculation of typical error and intraclass correlation. The pairwise comparisons revealed no consistent bias across tests for all three measures and there was no evidence of non-uniform errors (heteroscedasticity). When three trials were analysed together typical error expressed as a coefficient of variation was 4.1% for peak VO(2) 9.3% for cardiac output and 9.3% for stroke volume. Results analysed by sex revealed no consistent differences. The PhysioFlow method allows non-invasive, beat-to-beat determination of cardiac output and stroke volume which is feasible for measurements during maximal exercise in children. The reliability of the PhysioFlow falls between that demonstrated for Doppler echocardiography (5%) and CO(2) rebreathing (12%) at maximal exercise but combines the significant advantages of portability, lower expense and requires less technical expertise to obtain reliable results. PMID- 15827737 TI - Central processing overlap modulates P3 latency. AB - Two experiments examined the issue of the functional mechanisms exerting a modulatory effect on the latency of the P3. In experiment 1, using a psychological refractory period (PRP) paradigm, two sequential stimuli (T(1) and T(2)) were presented in each trial at varying stimulus onset asynchronies (SOAs), each requiring a speeded choice response. Substantial lengthening of the reaction time to T(2) was observed as SOA decreased (i.e., PRP effect). A systematic investigation of the T(2)-locked P3 component amplitude and latency was undertaken to discover whether either of these P3 parameters was correlated with the PRP effect. The results showed lengthening of the T(2)-locked P3 component latency as SOA was decreased, and, across subjects, a positive correlation between the PRP effect and P3 latency lengthening. No SOA-dependent P3 amplitude variation was observed. In experiment 2, the P3 component was measured under single-task conditions. P3 amplitude was higher under single-task than under dual task conditions, but no SOA-dependent latency variations were observed in this experiment. Overall, the results of both experiments support the notion that part of the processing reflected in P3 activity occurs at or after the locus of the PRP effect, thus suggesting strongly that central mechanisms are involved in P3 latency variations. PMID- 15827736 TI - Protection of human cerebral neurons from neurodegenerative insults by gene delivery of soluble tumor necrosis factor p75 receptor. AB - Apoptosis plays an important role in neuronal cell death in both chronic and acute human neurodegenerative diseases, including amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia, and human immunodeficiency virus (HIV) encephalopathy. We evaluated the ability of the extracellular binding domain of a dimeric tumor necrosis factor receptor (p75TNFR) to prevent neurotoxicity and death of human fetal cerebral neurons that were exposed in vitro to toxic agents known to be implicated in human neurological disorders, including tumor necrosis factor (TNFalpha) and the HIV proteins Tat and gp120. The extracellular domain of p75TNFR is capable of binding and neutralizing both soluble and transmembrane anchored TNFalpha. We efficiently transduced human neurons using adenoviral vectors expressing p75TNFR (Ad.p75TNFR) or a control gene (lacZ). Treatment of control cultures with the toxic agents TNFalpha, TNFalpha plus actinomycin D, or Tat and gp120, induced neurotoxic alterations and apoptotic death of neurons. By contrast, transduction of neurons with Ad.p75TNFR prevented apoptosis and cell death due to these agents. We conclude that viral vector transfer of the p75TNFR gene efficiently protects human neurons from TNFalpha-, Tat- or gp120-induced apoptosis and cell death. These results suggest that p75TNFR transduction of neurons by viral vectors could be therapeutically useful in the treatment of many human neurodegenerative diseases. PMID- 15827738 TI - Properties of saccades generated as a choice response. AB - Since Hick's original description, many subsequent studies have confirmed the logarithmic relationship that exists between response time and the number of alternatives (NA) for a choice response. In the present study a novel paradigm was used to quantify saccade response time as a function of NA. Normal subjects were required to make a saccade to the remembered location of a visual target whose color was specified by a centrally located cue. The paradigm thus required a stimulus-response transformation similar to that used by Hick. The results show that, when such a transformation was required, a logarithmic relationship was found for saccadic response time. The use of a color-to-location paradigm to study saccade choice response time produced an unexpected additional result that may provide insight into the neural organization of the saccadic system. When the number of alternative choice responses was large (4 or 8), subjects frequently made a two-saccade response instead of a single saccade to the correct location. The first movement in such a sequence was in the correct direction, but was hypometric. A second movement then followed which moved the eyes onto the correct location. These results suggest dynamic dissociations in the mechanisms underlying the triggering of saccades and the specification of their metrics. PMID- 15827739 TI - The right hand knows what the left hand is feeling. AB - The mislocalization profile, describing incorrect localization of faint tactile stimuli to different regions of the body, has been shown to provide insight into the processing of tactile stimuli. Interhemispheric somatosensory processing was examined in 15 subjects by studying the interference of left-hand stimulation on right-hand perception. In different conditions supra-threshold interference stimuli were applied to the left thumb or little finger either 200 or 500 ms prior to the application of a test stimulus on the right hand. Data show that interference stimuli applied to the left hand massively altered localization responses for stimuli applied to the right side. Stimulating the left thumb yielded an increased number of mislocalizations to the right thumb. Similarly, stimulating the left little finger caused a shift in localization responses towards the right ring finger. Results support the hypothesis that interaction of somatosensory information originating from different sides of the body follows a somatotopic organization. PMID- 15827740 TI - The nature and contribution of space- and object-based attentional biases to free viewing perceptual asymmetries. AB - Two experiments investigated the contribution of space- and object-based coordinates to previously reported leftward perceptual biases (pseudoneglect) at various locations across visual space. Neurologically intact participants (n = 34 and 27) made luminance discriminations between two left/right mirror-reversed luminance gradients (greyscales task), which were variously displaced around the midline in the participants' left and right hemispaces. The orientations of the stimuli were manipulated so that object- and space-based coordinates were congruent or incongruent. Experiment 1 confirmed the presence of a leftward object-based perceptual bias. The bias was moderated, however, by overattention to the more central stimulus. This central spatial effect could have resulted from the use of task-specific strategies, which were controlled by presenting the stimuli sequentially in Experiment 2. The findings of Experiment 1, a leftward object-based bias and a central spatial bias, were replicated. Overall, the results indicate a leftward object-based bias and a central spatial bias, both of which are relevant for the allocation of attention. The results are discussed with reference to a variety of models of the distribution of attention across space. PMID- 15827741 TI - Sex differences in cardiovascular and total mortality among diabetic and non diabetic individuals with or without history of myocardial infarction. AB - AIMS/HYPOTHESIS: We investigated the associations of type 2 diabetes and a history of myocardial infarction (MI) with coronary heart disease, stroke, cardiovascular and total mortality, and whether these effects are different in men and women. METHODS: Study cohorts included 51,735 Finnish men and women aged 25-74 years. Smoking status and the history of MI and type 2 diabetes were recorded, and height, weight, blood pressure and serum cholesterol levels were measured at baseline. Additional data on the occurrence of diabetes and MI prior to baseline survey were obtained from national hospital discharge and drug registers through computerised register linkage. Mortality data were obtained from the national mortality register. RESULTS: During a mean follow-up period of 17.2 years, 9,201 deaths were recorded. Hazard ratios (HRs) for coronary mortality were 2.1, 4.0 and 6.4 among men with diabetes alone, men with MI alone, and men with diabetes and MI, respectively, compared with men with neither diabetes nor a history of MI. The corresponding HRs among women were 4.9, 2.5 and 9.4, respectively. The equivalent HRs for cardiovascular and total mortality were 2.2, 3.3 and 6.0, and 1.8, 2.3 and 3.7 in men, and 4.4, 2.4 and 6.5, and 3.2, 1.7 and 4.4 in women, respectively. Diabetic men and women had similar total mortality rates, whereas total mortality rates were markedly higher among men in the other three diabetes-MI categories. CONCLUSIONS/INTERPRETATION: Diabetes and MI markedly increase cardiovascular and all-cause mortality. In women, the association between diabetes and mortality was stronger than that between MI and mortality, whereas the converse was true among men. PMID- 15827742 TI - The effects of the overexpression of recombinant uncoupling protein 2 on proliferation, migration and plasminogen activator inhibitor 1 expression in human vascular smooth muscle cells. AB - AIMS/HYPOTHESIS: Increased oxidative stress in vascular smooth muscle cells (VSMCs) has been implicated in the pathogenesis of accelerated atherosclerosis in patients with diabetes mellitus. Uncoupling protein 2 (UCP-2) is an important regulator of intracellular reactive oxygen species (ROS) production. We hypothesised that UCP-2 functions as an inhibitor of the atherosclerotic process in VSMCs. METHODS: Overexpression of human UCP-2 was performed in primary cultured human VSMCs (HVSMCs) via adenovirus-mediated gene transfer. Its effects on ROS production, AP-1 activity, plasminogen activator inhibitor 1 (PAI-1) gene expression, and cellular proliferation and migration were measured in response to high glucose and angiotensin II (Ang II) concentrations, two major factors in the pathogenesis of atherosclerosis in patients with diabetes and hypertension. Mitochondrial membrane potential and NAD(P)H oxidase activity were also measured. RESULTS: High glucose and Ang II caused transient mitochondrial membrane hyperpolarisation. They also significantly stimulated ROS production, NAD(P)H oxidase activity, mitochondrial membrane potential, AP-1 activity, PAI-1 mRNA expression, and proliferation and migration of HVSMCs. Adenovirus-mediated transfer of the UCP-2 gene reversed all of these effects. CONCLUSIONS/INTERPRETATION: The present study demonstrates that UCP-2 can modify atherosclerotic processes in HVSMCs in response to high glucose and Ang II. Our data suggest that agents increasing UCP-2 expression in vascular cells may help prevent the development and progression of atherosclerosis in patients with diabetes and hypertension. PMID- 15827743 TI - Widening of foot processes in normoalbuminuric adolescents with type 1 diabetes. AB - We compared renal morphology in normoalbuminuric adolescents with type 1 diabetes with age-and sex-matched controls. Renal morphological measurements in 46 unselected adolescents with diabetes for approx. 10 years were compared with those in 20 healthy kidney donors. Renal volume estimated by ultra-sound, renal function determined by clearances of inulin and para-aminohippurate, and long term mean HbA(1c) were measured in the patient group. Basement membrane thickness (BMT), mesangial matrix and capillary volume fractions per glomerulus [V(V)(matrix/glom), V(V)(cap/glom)] were significantly greater in patients than in controls (505 nm, 10.5%, 46.7% vs. 320 nm, 7.9% and 39.3%). The foot processes were wider in patients than in living donors (414 nm vs. 372 nm). There was no difference in mean kidney volume of patients with diabetes and healthy subjects. Glomerular filtration rate, body weight, and slit pore length density explained 60% of the variance in renal volume. After about 10 years' duration of type 1 diabetes, BMT, V(V)(matrix/glom), V(V)(cap/glom), and foot process width were greater in normoalbuminuric adolescents than in healthy controls. This increase in foot processes of normoalbuminuric patients has not been reported before. PMID- 15827744 TI - C1q nephropathy: features at presentation and outcome. AB - The study population comprised all 20 patients followed since 1990 through December 2004 at the Le Bonheur Children's Medical Center with diagnosis of C1q nephropathy (55% boys; 60% African Americans). All were aged under 18 years at biopsy (mean 11.2 years, 65% aged 11 or over); the youngest presented at age 10 months and progressed to end-stage renal disease at 14 months. None had clinical or laboratory features of systemic lupus erythematosis or membranoproliferative glomerulonephritis. Clinical features assessed at diagnosis were age, gender, blood pressure, history of macroscopic hematuria, urinary protein to creatinine ratio, serum creatinine, estimated glomerular filtration rate, renal histology, and pattern for immunofluorescent reactants. At the time of biopsy 40% had nephrotic syndrome and 30% nephrotic range proteinuria without nephrotic syndrome. Three patients with nephrotic syndrome also had chronic renal insufficiency at diagnosis. The most common histological feature was focal segmental glomerulosclerosis in 40%, but 30% had minimal change lesion. Four patients, all with nephrotic syndrome at diagnosis, progressed to end-stage renal disease. Of the 12 patients not presenting with nephrotic syndrome, none had chronic renal insufficiency at last follow-up. Kidney survival was 94% and 78% at 1 and 5 years, respectively, in all patients and 88% and 49% in those presenting with nephrotic syndrome. PMID- 15827745 TI - Association between the neuron-specific RNA-binding protein ELAVL4 and Parkinson disease. AB - Inflammatory processes have been implicated in the cascade of events that lead to nerve cell death. In the nervous system, a number of genes involved in inflammation pathways are regulated post-transcriptionally via the interaction of their mRNAs with specific RNA-binding Hu proteins, the vertebrate homologues of the Drosophila ELAV (for embryonic lethal abnormal vision). The gene encoding ELAVL4, a member of the Hu family of proteins, is located 2 Mb from the chromosome 1p linkage region peak for age-at-onset (AAO) of Parkinson disease (PD) (LOD = 3.41). Nine single-nucleotide polymorphisms (SNPs) in ELAVL4 were genotyped for 266 multiplex families (1,223 samples). Additional genotyping in 377 singleton families was performed for a subset of five SNPs (SNPs 1-5) that were not in linkage disequilibrium. SNP 2 (located in the first intron of ELAVL4) showed a strong significant association with AAO of PD (P = 0.006), and SNP 5 (a coding SNP in ELAVL4) showed a moderately significant association (P = 0.035). Haplotype analysis revealed that the A-C haplotype at SNPs 2 and 3 has the strongest significant association with AAO (P = 0.0001) among all combinations of two or three loci. The A-C haplotype remained significant for AAO after the inclusion of the C allele at SNP 5 to this haplotype (A-C-C haplotype, P = 0.00018). Although SNP 5 was found to associate with PD risk in the early-onset subset of PD families (at least one affected with AAO <40 years, 60 families), we believe that it is a by-product of its association with AAO. Taken together, these results suggest a potential role for ELAVL4 as a modifier gene for AAO of PD. PMID- 15827746 TI - Karyotypic evolution of a novel cervid satellite DNA family isolated by microdissection from the Indian muntjac Y-chromosome. AB - A minilibrary was constructed from DOP-PCR products using microdissected Y chromosomes of Indian muntjac as DNA templates. Two microclones designated as IM Y4-52 and IM-Y5-7 were obtained from negative screening of all three cervid satellite DNAs (satellites I, II, and IV). These two microclones were 295 and 382 bp in size, respectively, and shared approximately 70% sequence homology. Southern blot analysis showed that the IM-Y4-52 clone was repetitive in nature with an approximately 0.32-kb register in HaeIII digest. Sequence comparison revealed no similarities to DNA sequences deposited in the GenBank database, suggesting that the microclone sequences were from a novel satellite DNA family designated as cervid satellite V. A subclone of an Indian muntjac BAC clone which screened positive for IM-Y4-52 had a 3,325-bp insert containing six intact monomers, four deleted monomers, and two partial monomers. The consensus sequence of the monomer was 328 bp in length and shared more than 80% sequence homology with every intact monomer. A zoo blot study using IM-Y4-52 as a probe showed that the strong hybridization with EcoRI digested male genomic DNA of Indian muntjac, Formosan muntjac, Chinese muntjac, sambar deer, and Chinese water deer. Female genomic DNA of Indian muntjac, Chinese water deer, and Formosan muntjac also showed positive hybridization patterns. Satellite V was found to specifically localize to the Y heterochromatin region of the muntjacs, sambar deer, and Chinese water deer and to chromosome 3 of Indian muntjac and the X-chromosome of Chinese water deer. PMID- 15827747 TI - In vitro regeneration of Hagenia abyssinica (Bruce) J.F. Gmel. (Rosaceae) from leaf explants. AB - A protocol for shoot regeneration of Hagenia abyssinica (Bruce) J.F. Gmel. has been developed using leaf explants originating from in vitro seedlings and mature material. The explants were cultured on Murashige and Skoog medium containing various concentrations of alpha-naphthaleneacetic acid and thidiazuron (TDZ). Concentrations of TDZ lower than 1.0 microM promoted direct shoot regeneration, but higher concentrations promoted callus induction. Around 96-100% regeneration was obtained between 1.0 and 10 microM TDZ. The average number of shoots per explant at 1.0 microM TDZ was 8.4+/-4.8. Among the different explants used, the highest percentage of regeneration and shoots per explant was obtained from complete leaf explants. A significant (P< or =0.05) difference in regeneration capacity was observed among the five genotypes examined. The resulting shoots were multiplied on multiplication medium, rooted and acclimatised in a greenhouse. PMID- 15827748 TI - Novel keratin 5 and 14 gene mutations in patients with epidermolysis bullosa simplex from Poland. AB - Mutation analysis in keratins 5/14 (K5/14) had been performed in five Polish families with epidermolysis bullosa simplex (EBS) to extend genotype-phenotype correlation and to add to the mutation database. All the patients had been clinically classified into two subtypes of EBS; Weber-Cockayne (EBS-WC) and Dowling-Meara (EBS-DM) as well as one case of EBS with mottled pigmentation (EBS MP). DNA from patients and their family members was assessed for mutations in K5 or 14 using polymerase chain reaction amplification and subsequent direct sequencing. We identified four different missense mutations in K5 and one missense mutation in K14. Three of these are novel. Mutations associated EBS-DM resided in the highly conserved 20 amino acids end of the 1A domain in K5. Direct nucleotide sequencing of a case of EBS-MP revealed a heterozygous P25L mutation in K5. However, no genotype-phenotype correlation was identified in families with EBS-WC. The present study demonstrates the first series of molecular genetic data in EBS from Poland. PMID- 15827749 TI - Prosthetic replacement in secondary Girdlestone arthroplasty has an unpredictable outcome. AB - The functional outcome of hip replacement following a Girdlestone arthroplasty may be difficult to predict. We reviewed 39 hips in 39 patients with a minimum follow-up of 12 (range 12-208) months from re-implantation total hip arthroplasty. The patients were treated in one institution between 1983 and 2000, and their mean age at conversion was 65 (32-85) years. The main indication for the Girdlestone arthroplasty was peri-prosthetic infection. A post-operative complication occurred in 26 cases and in 17, surgical revisions were performed. The average Harris hip score (HHS) following conversion was 62 (24-93), and only three hips were graded as very good whereas 23 were graded as poor. Microbiological culture, patient age, duration of Girdlestone arthroplasty and the number of preceding surgical procedures did not correlate with the functional outcome after conversion. PMID- 15827750 TI - Inter-observer reliability in the assessment of heterotopic ossification: proposal of a combined classification. AB - We applied four published classifications for assessment of heterotopic ossification after total hip arthroplasty (Arcq, Brooker, DeLee and Della Valle [1, 2, 5, 6]. The average incidence of heterotopic ossification varied from 19.8% to 27.7%. The inter-observer reliability of the various classifications was determined by Cohen's Kappa statistic. Kappa values ranged from 0.897 for Arcq's to 0.814 for Brooker's classification. In order to increase the reliability and consistency, we propose a new classification system combining Brooker's and Della Valle's classifications. This new classification preserves the high reliability of Della Valle's system and is comparable to previous publications since it includes Brooker's criteria. PMID- 15827751 TI - Ileal pedicle grafting for esophageal replacement in children. AB - Reconstruction of the upper esophagus in small children remains a challenge. Free jejunal interposition as frequently used in adults is much less appropriate in children because of the limited vessel size. The use of a jejunal pedicle graft in children has been described, but gaining enough length may be a problem. A pedicle graft of terminal ileum may be a better option, but this technique has never been described. We report a child with esophageal atresia and distal fistula who had a very short upper esophageal pouch. Primary repair was impossible. The fistula was ligated and a gastrostomy created. A second attempt at anastomosis was not successful either, and a cervical esophagostomy was created. The child was fed by gastrostomy and received sham feeding orally. When the child was 10 months old, the upper esophagus was successfully reconstructed with a pedicle graft of terminal ileum. Postoperatively there was a limited leak of the proximal anastomosis, which healed spontaneously. The distal anastomosis had to be dilated on a few occasions. With a follow-up of 1 year, the child is eating well without gastrostomy supplementation. On imaging, the ileal pedicle graft looks somewhat tortuous but contracts nicely. We feel that ileal pedicle graft reconstruction of the esophagus should be part of the instrumentarium of pediatric surgeons dealing with esophageal reconstruction. PMID- 15827753 TI - [Current aspects of forensic law. From October 2004 to March 2005]. PMID- 15827752 TI - Mucosal expression of p21, p27, p53, Bcl-2, and bax after small bowel resection and autotransplantation in pigs. AB - Massive small bowel resection increases ileal villus height as part of normal adaptation. However, despite no gut loss, autotransplantation of the entire small intestine also increases ileal villus height. Our aim was to test whether similar modulation of enterocyte proliferation and apoptosis underpin these comparable increases in villus height. Fifteen pigs were randomly assigned for laparotomy (n=5), 75% proximal small bowel resection (n=5), or jejunoileal autotransplantation (n=5). Eight weeks postoperatively, full-thickness small bowel sections underwent routine immunohistochemistry for cell cycle inhibitors (p53, p21, and p27), antiapoptotic Bcl-2, and proapoptotic bax. The specimens were analyzed semiquantitatively, and the number of intensively positive epithelial cells for each group was compared from 20 digital images (0.32 mm(2)/image). Compared with laparotomy, small bowel resection decreased the number of p27-positive enterocytes in both jejunum and ileum, increased the number of bax-expressing cells in ileum, but decreased the number of bax expressing cells in jejunum. In contrast, compared with laparotomy, jejunoileal autotransplantation altered neither mucosal bax nor p27 expression. In all groups, Bcl-2 expression was similarly confined to inflammatory cells of the lamina propria, while both p53 and p21 were negative. We conclude that long-term alterations in the enterocytic expression of certain cell cycle and apoptosis markers (p27 and bax) accompany small bowel resection. These changes differ between the jejunum and the ileum and are not seen after whole small bowel autotransplantation. Therefore, increased ileal villus height after autotransplantation, despite resembling postresectional intestinal adaptation, is underpinned by different regulation of enterocyte proliferation and apoptosis. PMID- 15827754 TI - The novel calcineurin inhibitor ISA247: a more potent immunosuppressant than cyclosporine in vitro. AB - ISA247 is a novel cyclosporine analog. In this study we compare, in vitro, the effects of ISA247 on immune function with those of cyclosporine. Whole blood from cynomolgus monkeys (n = 5) was incubated with different concentrations of ISA247 or cyclosporine and stimulated with different mitogens in culture medium. Lymphocyte proliferation was assessed by [3H]-TdR incorporation assay and by flow cytometry. Flow cytometry was also used to assess production of intracellular cytokines by T cells and expression of T cell activation surface antigens. The concentration of drug necessary to attain 50% of the maximum effect (EC50) was subsequently calculated. EC50 values for ISA247 were lower than for cyclosporine, and the differences were statistically significant for lymphocyte proliferation, T cell cytokine production, and expression of all T cell activation surface antigens but one. We conclude that ISA247 suppresses diverse immune functions more potently than cyclosporine in vitro. PMID- 15827755 TI - Arterialisation of the portal vein as a model for the induction of hepatic fibrosis: description of microsurgical models in the rat. AB - Within the framework of liver transplantation, arterialisation of the portal vein in the case of non-recanalisable thrombosis has been reactivated. However, one of the consequences of this vascular reconstruction is the development of hepatic fibrosis. Clinical experience has shown that the development of fibrosis can be avoided by reducing portal inflow. We present, as a model for the induction of hepatic fibrosis, techniques of PVA, including transplantation. For PVA, several different techniques were used: the first with reduction of the portal inflow over a stent inserted in the right renal artery (PVA-B), the second with unrestricted flow using an aortic-portal segment (PVA-APS). The third technique was orthotopic liver transplantation with unrestricted portal arterialisation (OLTx-APS). Portal blood flow was measured with an ultrasonic flow probe. To determine the degree of hepatic fibrosis the amount of hydroxyproline was measured. Quantification of relative transcript levels of procollagen I was effected with real-time PCR using the TaqMan technology on a lightcycler instrument. The extracellular matrix was visualised with picro-sirius staining. Measurements with the ultrasonic probe showed a significant increase in flow rates, both with reduced (PVA-B) and unrestricted inflow (PVA-APS; OLTx-APS). The lowest survival rate (58%) was found in the group with unrestricted portal inflow. The reason for this was a high rate of thrombosis in the in the portal vascular tree (4 out of 12). In the OLTx-APS group four animals died within the first 3 postoperative days (69%), as a result of protracted postoperative shock. The overall survival rate was the highest (85%) in the group undergoing PVA with reduction of the portal inflow. PVA with unrestricted inflow was followed by a significant increase in extracellular collagen, which showed a clear correlation with the increase in the amount of hydroxyproline, the level of the mRNA for procollagen I and picro-sirius staining. With the operative PVA techniques presented herein, different arterial flow rates in the portal vein can be investigated. In our opinion these techniques represent an excellent animal model for studying the genesis of fibrosis and antifibrotic substances. By regulating the blood flow in the arterialised portal vein hepatic fibrosis can be reduced or even avoided. After a brief period of learning the microsurgical techniques, the surgeon can limit clamping times and achieve good results with these techniques. PMID- 15827756 TI - Human archival tissues provide a valuable source for the analysis of spatial genome organization. AB - Sections from archival formalin-fixed, paraffin wax-embedded human tissues are a valuable source for the study of the nuclear architecture of specific tissue types in terms of the three-dimensional spatial positioning and architecture of chromosome territories and sub-chromosomal domains. Chromosome painting, centromeric, and locus-specific probes were hybridized to tissue microarrays prepared from formalin-fixed paraffin wax-embedded samples of pancreas and breast. The cell nuclei were analyzed using quantitative three-dimensional image microscopy. The results obtained from non-neoplastic pancreatic cells of randomly selected individuals indicated that the radial arrangement of the chromosome 8 territories as well as their shape (roundness) did not significantly differ between the individuals and were in accordance with assumptions of a probabilistic model for computer simulations. There were considerable differences between pancreatic tumor and non-neoplastic cells. In non-neoplastic ductal epithelium of the breast there was a larger, but insignificant, variability in the three-dimensional positioning of the centromere 17 and HER2 domains between individuals. In neoplastic epithelial breast cells, however, the distances between centromere and gene domains were, on average, smaller than in non neoplastic cells. In conclusion, our results demonstrate the feasibility of studying the genome architecture in archival, formalin-fixed, paraffin wax embedded human tissues, opening new directions in tumor research and cell classification. PMID- 15827757 TI - DNA content heterogeneity in neuroblastoma analyzed by means of image cytometry and its potential significance. AB - The aim of this study was to determine the frequency and significance of the tumor DNA content heterogeneity in 33 previously untreated human neuroblastomas. We used image cytometry to selectively analyze neuroblasts by excluding karyorrhectic or stromal cells from cytometric measurements. DNA content heterogeneity with more than one clonal subpopulation on DNA histogram was found in 8 of 33 cases. Of these 8 cases, 4 showed MYCN amplification. Double labeling fluorescent in situ hybridization with probes for the centromeric region of chromosome 2 and MYCN gene was used to confirm the DNA content heterogeneity. DNA content heterogeneity was associated with poorer prognosis in this study (P<0.05). There was a significant correlation between euploidy (di- and tetraploidy) and worse prognosis, but only when heterogeneous neuroblastomas with euploid cell population were assigned to euploid tumors (P=0.006). Our results may explain the conflicting data in the literature regarding ploidy and suggest that DNA content heterogeneity and the presence of a euploid population may predict worse prognosis in neuroblastoma patients. PMID- 15827758 TI - Cardiopulmonary resuscitation and back injury in ambulance officers. AB - OBJECTIVE: Cardiopulmonary resuscitation (CPR) is frequently performed by allied health professionals, and, if undertaken in compromised positions, it may induce injury to the rescuer's spine. Back pain as an occupational symptom in manual workers has received considerable attention in the medical literature, but there is no report on the effect of CPR on back discomfort. This article reports a survey that explored the difficulties encountered by ambulance officers during the CPR procedure. METHODS: A questionnaire was sent to ambulance officers who had CPR experience through the Director and station officers-in-charge of the Fire Services Department in Hong Kong. RESULTS: Analysis of 318 completed questionnaires showed that the duration of the CPR procedure could last up to 32 min. Two hundred and ninety ambulance officers had experience in delivering CPR on a bed, and 60% of them often had to climb onto the bed with their legs overhanging to perform CPR. Eighty-nine percent were often required to turn their head or twist their back to look at the monitor during CPR. Almost 60% complained of always, and 36% sometimes, experiencing back discomfort during CPR. Only 4.5% had no experience of back discomfort during CPR. Seventy-six (24%) respondents suffered back injury, and as many as 62% of these 76 officers considered the cause of their back injury was related to CPR delivery. Nearly 50% of respondents reported that it was difficult to maintain balance and concurrently deliver CPR while travelling in the ambulance or when the patient was being transported from one location to another. CONCLUSIONS: Results of this survey suggest that there is a need to review the support given to ambulance officers to ensure safe administration of CPR, particularly during ambulance transport. PMID- 15827759 TI - The complex role of the chemokine receptor CCR2 in collagen-induced arthritis: implications for therapeutic targeting of CCR2 in rheumatoid arthritis. AB - CCR2 has been widely considered as a potential therapeutic target for autoimmune disease, particularly rheumatoid arthritis, and various CCR2 blocking agents have been developed, some of which have entered clinical trials. In this review, we examine the relevant information regarding the role of CCR2, and to a lesser extent of the closely related chemokine receptor CCR5, in the immunopathogenesis of collagen-induced arthritis, an animal model of rheumatoid arthritis. Experimental evidence showing that CIA is accelerated and exacerbated when CCR2 is genetically inactivated (knockout mice) or blocked with specific antibodies warrant additional investigations before the relevance of the findings in rodent models can be applied to human patients with RA. PMID- 15827761 TI - Agreement of expert judgment in causality assessment of adverse drug reactions. AB - BACKGROUND: Global introspection is, with operational algorithms and Bayes' theorem, one of the three main approaches used to assess the causal relationship between a drug treatment and the occurrence of an adverse event. OBJECTIVE: To analyze and compare the judgments of five senior experts using global introspection about drug causation on a random set of putative adverse drug reactions. METHODS: A random sample of 150 drug-effect pairs was constituted. For each pair, five senior experts had to independently assess the probability of drug causation from 0 to 1 by using a 100 mm visual analog scale (VAS). For analysis, those probabilities were secondarily split into seven levels of causality: excluded (0-0.05); unlikely (0.06-0.25); doubtful (0.26-0.45); unassessable/unclassifiable (0.46-0.55); plausible (0.56-0.75); likely (0.75 0.95); and certain (0.95-1). Agreement among the five experts was assessed using kappa coefficients (kappa). RESULTS: The overall agreement between experts was poor (kappa=0.20), although significantly different from chance, and varied according to the level of causality. It was lower for the unlikely, doubtful, unassessable/unclassifiable, and plausible categories (kappa=0.03, 0.03, -0.01, and 0.13, respectively) than for VAS extremes: excluded, likely, and certain (kappa=0.40, 0.32, and 0.30, respectively). CONCLUSION: This study confirms that experts express marked disagreements when assessing drug causality independently. The agreement rate was lower for intermediate levels of causality, especially when strong evidence was lacking for confirming or ruling out drug causality. Therefore, in a decision-making context, a step-by-step consensual approach such as the Delphi method seems necessary to make the assessment of such cases more reliable. PMID- 15827760 TI - The key role of apolipoprotein E in atherosclerosis. AB - Apolipoprotein E is a multifunctional protein that is synthesized by the liver and several peripheral tissues and cell types, including macrophages. The protein is involved in the efficient hepatic uptake of lipoprotein particles, stimulation of cholesterol efflux from macrophage foam cells in the atherosclerotic lesion, and the regulation of immune and inflammatory responses. Apolipoprotein E deficiency in mice leads to the development of atherosclerosis and re-expression of the protein reduces the extent of the disease. This review presents evidence for the potent anti-atherogenic action of apolipoprotein E and describes our current understanding of its multiple functions and regulation by factors implicated in the pathogenesis of cardiovascular disease. PMID- 15827762 TI - LGI1: a gene involved in epileptogenesis and glioma progression? AB - The leucine-rich, glioma inactivated gene 1 (LGI1) gene on human chromosome 10q24 was first identified as a candidate tumor suppressor gene for glioma. Surprisingly, mutations in LGI1 were also shown to cause an idiopathic epilepsy syndrome, autosomal dominant lateral temporal lobe epilepsy (ADLTE). LGI1 is one of the only two currently known non-ion channel genes whose mutations cause idiopathic epilepsy in humans. In this review we summarize the current data on structure and function of the LGI1 protein and discuss clinical aspects of ADLTE and their correlation with LGI1. We also propose that the evidence supporting the tumor suppressor role of LGI1 in malignant gliomas is weak and that further work is necessary to establish LGI1 role in glial cells. PMID- 15827763 TI - Diagnosis of Pelizaeus-Merzbacher disease: detection of proteolipid protein gene copy number by real-time PCR. AB - Duplication of the proteolipid protein gene (PLP1) is the most frequent cause of Pelizaeus-Merzbacher disease (PMD), a severe X-linked myelination disorder. We developed an assay for the detection of the PLP1 gene dosage by real-time quantitative PCR using the ABI Prism 7700 Sequence Detection System and the TaqMan chemistry. Copy number of the PLP1 gene was determined by the standard curve method using GAPDH as the reference gene. The assay was tested both on 50 normal controls and on 20 subjects whose PLP1 gene copy number was previously determined by quantitative fluorescent multiplex PCR. The procedure confirmed the expected results both on the male and female normal controls as well as on the 20 subjects previously tested. Ratios corresponding to the presence of one, two or three PLP1 gene copies, distributed in three non-overlapping ranges, were obtained by real-time PCR analysis. Subsequently, 29 DNA samples of putative PMD patients and possible female carriers, with unknown PLP1 gene dosage, were analysed. Five affected males carrying the PLP1 gene duplication and four female heterozygotes carrying three PLP1 gene copies were identified among them. The method is suitable for the identification of affected male patients and female carriers. Specific ranges are widely spaced, ensuring a correct assignment of the PLP1 gene copy number. PMID- 15827764 TI - Effect of subthalamic stimulation on mood state in Parkinson's disease: evaluation of previous facts and problems. AB - In an attempt to clarify the effect of deep brain stimulation (DBS) to the subthalamic nucleus (STN) on mood state, previous evidence and problems were evaluated through a systematic literature search. Twenty three articles reported the effect of STN DBS on mood state in Parkinson's disease (PD), and antidepressant, depressant, and mania-induced effects were reported in 16.7-76%, 2-33.3%, and 4.2-8.1% of the patients treated with STN DBS, respectively. Most articles reported larger subgroups showing antidepressant effects than those showing depressant effects. The average depression scale score of all subjects was improved or unchanged after STN DBS. Although there was a limitation due to the varied results, it was suggested that, in general, STN DBS had an antidepressant effect in PD. However, the studies reporting severe depressant symptoms, such as suicidal attempts, after STN DBS indicated the importance of careful attention to mood state as well as to motor symptoms after STN DBS. It may be crucial to reduce the variation in the results by, for example, the use of standardized protocols and the precise verification of the stimulated region in further investigations to address this issue. PMID- 15827765 TI - Atrophy of thigh muscles after meniscal lesions and arthroscopic partial menisectomy. AB - The purpose of this study was to investigate the atrophic pattern of the muscle groups and their individual muscles in the thigh after meniscal lesions and arthroscopic partial menisectomy. A total of 32 individuals (17 men and 15 women) who underwent arthroscopic knee surgery participated in this study. Their operated and non-operated thighs were scanned by magnetic resonance imaging to determine the volume of the quadriceps (QF), hamstring (HM), and adductors (AD). Compared with the non-operated limb, the volume of the QF was significantly lower in the operated limb; however, no significant difference was observed in the HM and AD. The volume of individual muscles of the QF, i.e. the rectus femoris (RF), vastus lateralis (VL), vastus intermedius (VI), and vastus medialis (VM), in the operated limb was significantly lower than the volume of those in the non operated limb (P<0.01, all). Although the relative change in the VM was significantly higher than that of the RF (P<0.05), specific atrophy was not found among four individual muscles in the QF. We concluded that meniscal lesions and partial menisectomy induce atrophy in the QF only in the thigh, and that no specific atrophy, e.g. VM, seemed to occur within the individual muscles in the QF. PMID- 15827766 TI - In vivo kinematics of the ACL-deficient limb during running and cutting. AB - Anterior cruciate ligament-deficient (ACLD) knee kinematics during high-demand activities are poorly understood. We have devised a new method, using gait analysis, to more accurately assess 3-D in vivo kinematics of the knee. This has enabled us to report on how knee kinematics are altered after ACL rupture, during running and cutting. Fifteen unilaterally ACLD subjects were assessed using a 12 camera 100 Hz VICON motion analysis system. Simultaneous electromyographical (EMG) recordings were used to assess the role of the sensorimotor system in knee joint stability. All subjects were able to perform demanding cutting activities without experiencing symptoms of instability. We found that running produces fundamentally different kinematic patterns to those seen during walking. Tibiofemoral translation in the anteroposterior plane is controlled to within normal limits. Conversely, coronal translation and rotation are poorly controlled. We found that the injured leg was maintained in greater extension during the stance phase of all running activities studied and that the quadriceps muscle was active for longer during this period. We believe that low-demand activities, such as walking, do not reproduce kinematics relevant to ACLD instability and that future investigations into functional instability in the ACLD knee should focus on coronal and rotational displacements. PMID- 15827767 TI - A phase I study of doxifluridine combined with weekly paclitaxel for metastatic gastric cancer. AB - PURPOSE: Based on the synergistic effect in preclinical studies, a phase I clinical trial for the combination of paclitaxel and doxifluridine (an intermetabolite of capecitabine) was performed to determine the recommended dose for the treatment of patients with metastatic gastric cancer. METHODS: The dose of paclitaxel was increased from 60 mg/m2 at level 1 to 90 mg/m2 at level 5. It was administered as a 1-h infusion on days 1 and 8. The dose of doxifluridine was fixed at 600 mg/m2 per day up to level 3, and escalated to 800 mg/m2 per day at levels 4 and 5. It was administered orally for 2 weeks. The treatment was repeated every 3 weeks. RESULTS: A total of 28 patients were enrolled. No dose limiting toxicity (DLT) was observed at levels 1 and 2 (paclitaxel 70 mg/m2). A DLT of grade 4 neutropenia lasting for more than 4 days was observed in one patient at level 3 (paclitaxel 80 mg/m2). In addition, the first five of six patients in this group experienced grade 3 neutropenia during the first treatment cycle. A further six patients were added in order to confirm the safety of this dosage level, and no more DLTs except for grade 3 nausea in one patient were observed in the second cohort. No DLT was seen in three patients at level 4 (paclitaxel 80 mg/m2). DLTs (grade 3 neuropathy in one patient and a treatment delay of the second cycle for more than 1 week due to grade 3 neutropenia in another) were observed in two out of six patients at level 5 (paclitaxel 90 mg/m2), and this dose level was determined as the maximum tolerated dose. The tumor response rate was 42% (95% confidence interval 20-67%) in 19 patients with measurable lesions. CONCLUSIONS: The recommended dose was determined as 80 mg/m2 of paclitaxel (days 1 and 8) and 800 mg/m2) of doxifluridine (days 1-14) every 3 weeks. The results of this phase I study are encouraging and a phase II trial is thus warranted. PMID- 15827768 TI - Less is (usually) more: when is amputation appropriate for treatment of extremity soft tissue sarcoma? PMID- 15827769 TI - Reducing the need for sentinel node procedures by ultrasound examination of regional lymph nodes. PMID- 15827770 TI - Simplification of breast cancer surgery. PMID- 15827771 TI - "Axillary recurrence after a negative sentinel node biopsy": editorial comment. PMID- 15827772 TI - The indications for and the prognostic significance of amputation as the primary surgical procedure for localized soft tissue sarcoma of the extremity. AB - BACKGROUND: The indications for primary amputation of a localized soft tissue sarcoma (STS) of the extremity are not well defined in the literature. However, it has been suggested that patients who require an amputation to treat an STS are at increased risk for developing metastases. We categorized the main indications for primary amputation in our patient population and compared their oncological outcome with the outcome of patients who underwent limb-sparing surgery. METHODS: 413 consecutive patients treated surgically at a single center for primary, nonmetastatic, deep, intermediate-, or high-grade STS of the extremity were reviewed. Indications for primary amputation were identified. Demographics and outcomes were compared between the amputation and limb-salvage groups. Multivariate Cox model analysis was used to identify independent risk factors for systemic relapse. RESULTS: Twenty-five (6%) of 413 patients with STS underwent primary amputation: they were older (P = .05), had larger tumors (P = .001), and had a significantly greater risk of developing metastatic disease than patients who underwent limb-sparing procedures (P = .008). However, multivariate analysis demonstrated that the only independent predictors of systemic relapse were tumor size (P = .0001) and tumor grade (P = .0001). Primary amputation was not an independent risk factor for metastatic disease. CONCLUSIONS: The decision to perform a primary amputation for an STS of the extremity is based on the location and local extent of the tumor, and the expected function of the extremity after tumor resection. The higher risk of metastases for patients who require primary amputation is accounted for by independent risk factors associated with their tumors--predominantly large tumor size. PMID- 15827773 TI - Ultrasound examination of sentinel nodes in the initial assessment of patients with primary cutaneous melanoma. AB - BACKGROUND: The value of targeted high-resolution ultrasound (US) examination in detecting sentinel lymph node metastases in patients with newly diagnosed primary cutaneous melanomas has not yet been fully evaluated. The aim of this study was to determine the threshold size of metastatic melanoma deposits in SLNs able to be detected by targeted US examination before initial melanoma surgery. METHODS: A total of 304 patients presenting with primary cutaneous melanomas had SLNs identified by lymphoscintigraphy and then examined in situ by the same physician with high-resolution US. Within 24 hours, the SLNs were removed for histopathologic assessment of sections stained conventionally and with immunohistochemical markers for S100 protein and HMB45 antigen. RESULTS: Metastatic disease was present in SLNs from 33 node fields in 31 patients. The US results in seven of these cases were suggestive of metastatic disease. Twenty-six node fields contained positive nodes not detected by US. Undetected deposits had diameters <4.5 mm. CONCLUSIONS: These results suggest that a targeted US examination of SLNs can detect metastatic melanoma deposits down to approximately 4.5 mm in diameter. However, most metastatic melanoma deposits in SLNs are considerably smaller than this at the time of initial staging, and US therefore cannot be considered cost-effective in this setting. PMID- 15827774 TI - Can sentinel lymph node biopsy be omitted in patients with favorable breast cancer histology? AB - BACKGROUND: The widespread use of sentinel lymph node biopsy (SLNB) to replace axillary dissection has broadened the indications for axillary staging in breast cancer. Recent studies have demonstrated a finite risk of lymphedema and sensory morbidity associated with SLNB. We undertook this study to determine whether SLNB could be omitted in clinically node-negative patients with favorable-histology breast cancer. METHODS: We conducted a retrospective review of a prospective database of SLNBs performed at Memorial Sloan-Kettering Cancer Center from 1996 to 2003 to determine the incidence of lymph node metastases by histological subtype. For the favorable subtypes, the patient's age, tumor size, estrogen receptor status, lymphovascular invasion, nuclear grade, and histological grade were compared by nodal status to determine their predictive value. RESULTS: A total of 196 cases with favorable breast cancer subtypes were identified with a 4.1% (8 of 196) sentinel lymph node (SLN) positivity rate. Each of the histological subtypes included patients with positive SLNs, with the exception of adenoid cystic (n = 4) and secretory (n = 1) breast carcinoma, which were quite rare in our series. When compared by nodal status, the patient's age, tumor size, estrogen receptor status, lymphovascular invasion, nuclear grade, and histological grade failed to predict those with positive SLNs. CONCLUSIONS: Patients with favorable breast cancer histology have a small risk of axillary SLN metastases. The use of SLNB in these patients should be individualized, taking into consideration the small incidence of axillary metastases and the risks and benefits associated with the SLN procedure. PMID- 15827775 TI - Axillary recurrence after a negative sentinel node biopsy for breast cancer: incidence and clinical significance. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) carries the inherent risk of approximately 5% false-negative sampling. Undetected tumor-positive nodes of clinical importance are those that lead to axillary recurrence. This survey aims at clarifying the extent of this problem in current practice and literature. METHODS: In a regional teaching hospital, 696 consecutive breast cancer patients underwent SLNB between January 1998 and July 2003, and data were entered in a prospective database. PubMed and the Cochrane library were searched for a systematic review of the literature. Thirteen studies dealt with the follow-up of a cohort of sentinel lymph node (SLN)-negative patients or presented a case report. RESULTS: The SLN identification rate was 97.1%. The SLN was tumor free in 439 (65%) of the 676 patients. After a median follow-up of 26 months, axillary recurrence was detected in 2 of 439 patients 4 and 27 months after the SLNB. The incidence of clinically apparent false-negative SLNB is .46%. The systematic review resulted in 3184 SLNB-negative patients with a median follow-up of 25 months. Axillary recurrence occurred in eight patients after a median of 21 months. The axillary recurrence rate in the literature is .25%. One third of these patients present with synchronous systemic metastases. CONCLUSIONS: Axillary recurrences after a negative SLNB occur, but at a much lower rate than would be expected on the basis of historical figures and the false-negative SLN findings. The natural history of axillary relapse after negative SLNB resembles the locoregional recurrence of breast cancer. PMID- 15827776 TI - Axillary recurrence after sentinel node biopsy. AB - BACKGROUND: Sentinel node biopsy (SNB) has evolved as the standard of care in the surgical staging of breast cancer. This technique is accurate for surgical staging of axillary nodal disease. We hypothesized that axillary recurrence after SNB is rare and that SNB may provide regional control in patients with microscopic nodal involvement. METHODS: With institutional review board approval, SNB was performed with peritumoral injection of 99mTc-labeled sulfur colloid. From 1996 to 2003, 1167 patients were entered into a prospective cancer database after surgical therapy; 916 patients consented to long-term follow-up. Fifty-two patients (5.7%) did not map successfully and were excluded, leading to a study population of 864 patients. The median follow-up was 27.4 months (range, 1-98 months). RESULTS: The median number of sentinel nodes harvested was 2, and 633 (73%) patients had negative sentinel nodes. Thirty (4.7%) of those sentinel node negative patients underwent completion axillary dissection, whereas 592 (94%) patients were followed up with observation. A total of 231 (27%) had positive sentinel nodes: 158 (68%) of these patients underwent completion axillary dissection, and 73 (32%) were managed with observation alone. Two (.32%) patients who were sentinel node negative had an axillary recurrence; one of these patients had undergone completion axillary dissection. No patient in the observed sentinel node-positive group had an axillary recurrence (odds ratio, .37; P = .725). CONCLUSIONS: On the basis of a median follow-up of 27.4 months, axillary recurrence after SNB is extraordinarily rare regardless of nodal involvement, thus indicating that this technique provides an accurate measure of axillary disease and may impart regional control for patients with node-positive disease. PMID- 15827777 TI - Beyond the clinical trials: how often is sentinel lymph node dissection performed for breast cancer? AB - BACKGROUND: Sentinel lymph node dissection (SLND) has been shown to be a reasonable treatment option for early-stage breast cancer. Until recently, SLND was limited to clinical trials. Because this technique is now offered outside of trials, its prevalence is unknown. METHODS: All patients with stage I or II breast cancer in the Surveillance, Epidemiology, and End Results national cancer registry (1998-2000) were evaluated. Data were collected for demographics, tumor characteristics, surgical resection, lymph node evaluation (SLND or complete axillary dissection), registry site, and year of diagnosis. Multivariate regression analysis was performed to identify predictors for receiving SLND. RESULTS: A total of 54,772 patients diagnosed with breast cancer had undergone surgical lymph node evaluation; 27.2% patients with stage I disease underwent SLND, as compared with 22.7% for stage II. Older patients and minority groups were less likely to receive SLND. Receipt of SLND varied by registry site (7.9% 32.7%). Multivariate regression showed that older patients had lower odds of receiving SLND (60-69 years: odds ratio, .73; P < .0001) as compared with younger patients. Additionally, blacks, Hispanics, and Asians had lower odds of receiving SLND (odds ratio of .64, .58, and .80, respectively; P < .0001). SLND use increased over the 3 years in the study (P < .0001). CONCLUSIONS: This population based analysis showed relatively infrequent use of SLND for early-stage breast cancer. These results suggest a slow transition of this procedure from clinical trials into the community. Future work should be targeted at improving the rate at which patients receive this procedure, particularly for elderly and minority groups and low-use regions. PMID- 15827778 TI - A predictive model for the development of hormone-responsive breast cancer. AB - BACKGROUND: Effective therapies to reduce the risk of hormone-sensitive breast cancers (ER or PR positive) exist. Available models predict the risk of breast cancer without addressing hormone receptor status. The purpose of this study was to identify risk factors predictive of the development of hormone-sensitive cancers. METHODS: A total of 1285 invasive breast cancers in 1263 women were identified from a prospectively maintained database. Risk factors were compared for ER+ and ER- cancers by using Fisher's exact test. RESULTS: Models were developed for premenopausal and postmenopausal women. In premenopausal women, white race, age at menarche < 12 years, and nulliparity or age at first birth > 20 years were used. The risk of ER+ cancer increased from 67.7% with 0 variables to 83.8% with all three (P = .013). In postmenopausal women, white race and a history of estrogen therapy were used. With none of the variables present, the incidence of ER+ cancer was 70.0%; it was 77.6% with one variable and 85.4% with both variables (P = .002). In postmenopausal women, variables predicted significant differences in hormone sensitivity only for those aged < or = 60 years. In the subset of women with information on alcohol use, adding this variable to the model improved the prediction of hormonal status. CONCLUSIONS: Our findings, if prospectively validated, may help identify those who would obtain the greatest benefit from hormonal chemoprevention strategies for breast cancer risk reduction. PMID- 15827779 TI - Ankle instability after vascularized fibular harvest for tumor reconstruction. AB - BACKGROUND: The vascularized fibular graft is an important tool in the reconstruction of defects caused by resections of orthopedic tumors. Children often undergo this form of reconstruction, but there is little information about the complications after vascularized fibular graft harvest in this age group. METHODS: We present a series of 32 patients who underwent this procedure to reconstruct an extremity in our institution. There were 12 children and 20 adults. RESULTS: The residual distal fibula was significantly longer in adults as compared with children (P < .048). Among children, 3 of 11 undergoing the procedure developed ankle instability, in distinction to adults, none of whom developed this complication (P < .041). This reflects a disruption of normal ankle function that develops in skeletally immature patients with a short residual fibula but not in patients with a longer residual fibula (P < .008). When the sum of patient age in years and residual fibula length in centimeters was less than 16, 3 of 6 patients developed deformity, in contrast to no deformity developing in the remaining 23 when the sum was >16 (P < .004). Adults were more likely to develop pain than instability. CONCLUSIONS: Our series suggests that children with an age-length sum <16 should be considered for prophylactic tibiofibular synostosis creation. PMID- 15827780 TI - Long-term survival of peritoneal carcinomatosis of colorectal origin. AB - BACKGROUND: Peritoneal carcinomatosis of colorectal cancer is probably best treated by cytoreduction and hyperthermic intraperitoneal chemotherapy (HIPEC). In The Netherlands Cancer Institute, this treatment has been performed since 1995. The long tradition of this treatment enabled us to study long-term survival in detail. METHODS: Between 1995 and 2003, 117 patients were treated by cytoreduction and HIPEC. The aim of the cytoreduction was to remove all visible tumor. After the cytoreduction, the abdomen was perfused with mitomycin C (35 mg/m2) at 40 degrees C to 41 degrees C for 90 minutes. Survival was calculated by the Kaplan-Meier method. Survival was also analyzed for the following subgroups: no residual tumor, residual tumor < or = 2.5 mm, and more residual tumor. Hazard ratios for each of the seven abdominal regions were calculated to determine the influence on survival. RESULTS: The median survival was 21.8 months. The 1-, 3-, and 5-year survival rates were 75%, 28%, and 19%, respectively. The Kaplan-Meier curve reached a plateau of 18% at 54 months. In 59 patients a complete cytoreduction was achieved, and in 41 patients there was minimal residual disease. The median survival of these patient groups was 42.9 and 17.4 months, respectively. When gross macroscopic tumor was left behind, as was the case in 17 patients, the median survival was 5 months. Involvement of the small bowel before cytoreduction was associated with poorer outcome. CONCLUSIONS: Cytoreduction followed by HIPEC showed a median survival of 21 months. From 3 years on, a consistent group of 18% of patients stayed alive. PMID- 15827781 TI - Lymph node metastasis in epithelial malignancies of the appendix with peritoneal dissemination does not reduce survival in patients treated by cytoreductive surgery and perioperative intraperitoneal chemotherapy. AB - BACKGROUND: Peritoneal dissemination of appendiceal malignancy combined with regional lymph node metastasis is an unusual combination of patterns of cancer dissemination. METHODS: A database of 501 appendiceal malignancy patients, all with documented peritoneal seeding, was used to identify 25 patients with involvement of the regional lymph nodes. All patients were uniformly treated with cytoreductive surgery plus perioperative intraperitoneal chemotherapy with mitomycin C and 5-fluorouracil. The clinical and pathologic features of the lymph node-positive patients were compared with those of the lymph node-negative patients. The effect of regional lymph node involvement on survival was determined. Within the group of lymph node-positive patients, clinical and pathologic features were tested for their effect on survival. RESULTS: When compared with patients with no apparent lymph node positivity, patients with positive lymph nodes were more likely to have an acute abdomen as the initial presentation (P < .001). The intestinal (nonmucinous) histological type was more common (P < .001), and the disseminated peritoneal adenomucinosis histology was less common (P < .001). Survival with the aggressive treatment strategy used in these patients was not different for lymph node-positive as compared with lymph node-negative patients (P = .15 by univariate and P = .38 by multivariate analysis). CONCLUSIONS: Appendiceal malignancy with dissemination to the lymph nodes has a more acute onset and a more frequent nonmucinous histology. With aggressive treatment strategies, lymph node-positive patients did not show a statistically significantly diminished survival. PMID- 15827782 TI - Extent of thyroidectomy is not a major determinant of survival in low- or high risk papillary thyroid cancer. AB - BACKGROUND: The optimal extent of thyroidectomy for papillary thyroid cancer (PTC) is controversial. Our objective was to evaluate the effect of total thyroidectomy or partial thyroidectomy on survival in low- and high-risk patients. METHODS: The Surveillance, Epidemiology, and End Results database was used to identify PTC patients who underwent thyroidectomy. The independent effects of age, distant metastases, extrathyroidal extension, tumor size, sex, lymph node metastases, radioactive iodine use, and extent of thyroidectomy on survival were analyzed for low- and high-risk PTC. RESULTS: There were 4402 (81%) low-risk and 1030 (19%) high-risk patients; 84.9% underwent total thyroidectomy. The 5- and 10-year survival were 95% and 89% in the low-risk patients and 84% and 73% in the high-risk patients, respectively (P = .001). In the low-risk patients, 10-year survival after total thyroidectomy was 89%, compared with 91% after partial thyroidectomy (adjusted hazard ratio for death, 1.73; 95% confidence interval, 1.28-2.33; P < .001); older age, male sex, larger tumor, lymph node metastases, and lack of radioactive iodine were associated with higher mortality. In the high-risk patients, 10-year survival after total thyroidectomy was 72%, compared with 78% after partial thyroidectomy (adjusted hazard ratio for death, 1.46; 95% confidence interval, .89-2.40; P = .14); older age, distant metastases, larger tumors, and lack of radioactive iodine were associated with higher mortality. CONCLUSIONS: Survival of patients with PTC was not significantly influenced by the extent of thyroidectomy. The survival after partial thyroidectomy was similar to total thyroidectomy within both the low- and high risk prognostic groups. PMID- 15827784 TI - Pelvic reconstruction after abdominoperineal resection: is it worthwhile? PMID- 15827785 TI - Rectal cancer: can we throw away the scalpel? PMID- 15827786 TI - Intraductal papillary mucinous neoplasms of the pancreas: a plea for prospective differentiation between main-duct and side-branch tumors. PMID- 15827787 TI - Does size matter most? Reassessing clinical staging for pancreatic cancer. PMID- 15827788 TI - In vivo sentinel lymph node mapping in lung cancer. PMID- 15827789 TI - Rectus flap reconstruction decreases perineal wound complications after pelvic chemoradiation and surgery: a cohort study. AB - BACKGROUND: A major source of morbidity after abdominoperineal resection (APR) after external beam pelvic radiation is perineal wound complications, seen in up to 66% of cases. Our purpose was to determine the effect of rectus abdominus myocutaneous (RAM) flap reconstruction on perineal wound morbidity in this population. METHODS: The study group consisted of 19 patients with anorectal cancer treated with external beam pelvic radiation followed by APR and RAM flap reconstruction of the perineum. A prospectively collected database was queried to identify a control group (n = 59) with anorectal cancer treated with similar radiation doses that subsequently underwent an APR without a RAM flap during the same time period. Comparison of percentages was performed with a two-sided Fisher's exact test, and comparison of means was performed with Wilcoxon's test. RESULTS: Perineal wound complications occurred in 3 (15.8%) of the RAM flap patients and 26 (44.1%) of the control patients (P = .03). The incidence of other complications was not different between groups (42.1% vs. 42.4%; P = .8). Despite an increased number of anal squamous tumors, an increased vaginectomy rate, increased use of intraoperative radiotherapy, and an increased proportion of cases with recurrent disease, the flap group had a significantly lower rate of perineal wound complications relative to the control group. CONCLUSIONS: Perineal closure with a RAM flap significantly decreases the incidence of perineal wound complications in patients undergoing external beam pelvic radiation and APR for anorectal neoplasia. Because other complications are not increased, RAM flap closure of the perineal wound should be strongly considered in this patient population. PMID- 15827790 TI - Relationship between pathologic T-stage and nodal metastasis after preoperative chemoradiotherapy for locally advanced rectal cancer. AB - BACKGROUND: We investigated the relationship between pathologic T-stage and mesorectal metastases after preoperative chemoradiotherapy (CRT) for clinical stage II to III rectal carcinoma. METHODS: The records of consecutive patients with clinical stage II to III carcinoma of the mid or low rectum who underwent surgery after CRT were reviewed. Indications for preoperative CRT were cancer up to 11 cm from the anal verge, Eastern Cooperative Oncology Group performance status of 0 to 2, age 18 to 75 years, and clinical tumor-node-metastasis stage II or III. RESULTS: The study group consisted of 235 patients (148 men and 87 women; median age, 61 years). The pretreatment tumor-node-metastasis stage was as follows: I, n = 1; II, n = 96; and III, n = 138. Radiotherapy was delivered at a median dose of 50.4 Gy. A pathologic complete response on the rectal wall was found in 24% of patients, and nodal metastases were found in 20% of patients. According to the pT stage, the rate of node positivity was 2% for pT0, 15% for pT1, 17% for pT2, 38% for pT3, and 33% for pT4 cases. At multivariate analysis, the best model for predicting pathologic node involvement included young age, positive pretreatment N status, and pT status. On considering pT stage alone, the odds ratio was in the region of 10 for pT1/2 and >20 for pT3/4 patients. CONCLUSIONS: In patients with pT0 after preoperative CRT for clinical stage II to III mid or low rectal cancer, the risk of nodal metastases is very low. More conservative surgery (local excision) may be considered in these cases. PMID- 15827791 TI - Prospective evaluation of quality of life of patients receiving either abdominoperineal resection or sphincter-preserving procedure for rectal cancer. AB - BACKGROUND: Study results on quality of life (QoL) between patients receiving an anterior resection (AR) or abdominoperineal resection (APR) for rectal cancer vary greatly. A main reason is grounded in unequal methodology. The aims of this study were to assess differences in perceived QoL over time among patients treated with AR or APR with a recommended study design and methodology. METHODS: In a prospective study, the European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire C30 and a tumor-specific module were administered to patients with rectal cancer before surgery, at discharge, and 3, 6, and 12 months after the operation. Comparisons were made between patients receiving an AR and those receiving an APR. RESULTS: Two hundred forty-nine patients were included; 46 patients received an APR and 203 an AR. QoL data were available for 212 patients, of which 112 were female and 100 male. No differences in the distribution of age, sex, or tumor stage were observed between groups. EORTC function scales showed no significant differences, including body image scales, between patients receiving an AR and those receiving an APR. In symptom scores, AR patients had more difficulty with diarrhea and constipation, whereas patients with APR experienced more impaired sexuality and pain in the anoperineal region. At discharge, patients receiving an AR were more confident about their future. CONCLUSIONS: QoL in patients receiving an AR and those receiving an APR is not different. Although patients with APR experience more impaired sexuality, patients receiving an AR experience decreases in QoL because of impaired bowel function. PMID- 15827792 TI - Multicenter analysis of clinicopathologic features of intraductal papillary mucinous tumor of the pancreas: is it possible to predict the malignancy before surgery? AB - BACKGROUND: Despite recently increasing numbers of reports on intraductal papillary mucinous tumors (IPMTs), difficulties still remain in terms of diagnosis, treatment, and prognosis. The purpose of this multicenter study was to evaluate the clinicopathologic features of IPMT in Korea and to suggest predictive criteria for malignancy in IPMT. METHODS: We retrospectively reviewed the clinicopathologic data of 208 patients who underwent operations for IPMT between 1993 and 2002 at 28 institutes in Korea. RESULTS: Of the 208 patients (mean age, 61 years), 147 were men and 61 were women. A total of 124 patients underwent pancreatoduodenectomy, 42 underwent distal pancreatectomy, 17 underwent total pancreatectomy, and 25 underwent limited pancreatic resection. There were 128 benign cases (adenoma, n = 62; borderline, n = 66) and 80 malignant cases (noninvasive, n = 29; invasive, n = 51). A significant difference in 5-year survival was observed between the benign and malignant groups (92.6% vs. 65.3%; P = .006). Of the six factors (age, location, duct dilatation, mural nodule, main duct type, and tumor size) that showed statistical differences by univariate analysis between the benign and malignant groups, three were significant by multivariate analysis--namely, mural nodule (P = .009), tumor size (P = .023), and a dilated duct size (P = .010). CONCLUSIONS: A significant proportion of IPMTs are malignant, although the overall prognosis of IPMT is superior to that of ordinary pancreatic cancer. Radical surgery is recommended for IPMT with the predictors of malignancy: mural nodule, tumor size (> or =30 mm), and dilated duct size (> or =12 mm). PMID- 15827793 TI - Hepatic resection at a major community-based teaching hospital can result in good outcome. AB - BACKGROUND: The relationship between volume and outcome has been established in the literature for several complex surgical procedures. Improved outcome has been suggested at high-volume hospitals or with high-volume surgeons. METHODS: The objective of this study was to evaluate the experience of a low-volume hospital with major liver resections. The setting of the study was a community-based teaching hospital with a surgical residency training program. RESULTS: A total of 46 major liver resections were performed between January 1992 and December 2002. Procedures performed were hepatic lobectomies (n = 15; right, n = 11; left, n = 4), trisegmentectomies (n = 5; right, n = 3; left, n = 2), segmentectomies (n = 16; left lateral, n = 12; right posterior, n = 4), and wedge resections (n = 10). Operations were performed by 14 different surgeons; however, 23 operations were performed by 1 surgeon. Sixteen patients (34%) developed 23 complications. The average length of hospital stay was 9.7 days. There were no 30-day postoperative mortalities. Out of 46 patients who underwent major liver resection over the last 10 years, 13 patients are still alive. Overall survival ranged from 3 to 84 months, with a median survival of 30.6 months. The actual 5-year survival was 36% (8 of 22) for all patients operated on >5 years ago, and the actual 2-year survival was 61% (20 of 33). CONCLUSIONS: Major liver resection can be performed safely with low rates of morbidity and operative mortality with careful selection of patients at a low-volume community-based teaching hospital. PMID- 15827794 TI - Isolated hepatic perfusion for the treatment of patients with colorectal cancer liver metastases after irinotecan-based therapy. AB - BACKGROUND: Irinotecan given with 5-fluorouracil and leucovorin is currently used as first-line therapy for patients with metastatic colorectal cancer (CRC). However, the response duration is <1 year, and second-line systemic chemotherapy has limited efficacy. We analyzed the efficacy of isolated hepatic perfusion (IHP) for patients with progressive CRC liver metastases after irinotecan. METHODS: Between March 1993 and February 2003, 124 patients with CRC liver metastases underwent IHP on institutional review board-approved protocols. The overall treatment mortality was 4% (5 of 124). Twenty-five patients (10 women and 15 men; mean age, 53 years) were identified who had progressive liver metastases by carcinoembryonic antigen, imaging studies, or both after irinotecan. A 1-hour hyperthermic IHP (mean hepatic temperature, 40.0 degrees C) with melphalan 1.5 mg/kg (mean total dose, 100 mg) was administered via laparotomy. Perfusion with an oxygenated extracorporeal circuit was established with inflow via a cannula in the gastroduodenal artery and common hepatic artery inflow occlusion. Outflow was via a cannula in an isolated segment of the inferior vena cava. During IHP, portal and inferior vena caval flow were shunted to the axillary vein. Patients were assessed for radiographical response, recurrence pattern, and survival. RESULTS: The mean number of prior irinotecan cycles in 25 patients was 6 (range, 2-14), and it was given primarily as second-line therapy. The median number of liver metastases before IHP was 10 (range, 1-50), and the median percentage of hepatic replacement by tumor was 25%. The mean operative time was 9 hours (range, 6-12 hours), and the median hospital stay was 11 days (range, 8-76 days). There was 1 complete response and there were 14 partial responses in 25 patients (60%), with a median duration of 12 months (range, 5-35 months). Disease progressed systemically in 13 of 25 patients at a median of 5 months (range, 3-16 months). The median overall survival was 12 months (range, 1-47 months), and the 2-year survival was 28%. CONCLUSIONS: For patients with progressive CRC liver metastases after irinotecan, IHP has good efficacy in terms of response rate and duration. Continued evaluation of IHP with melphalan as second-line therapy in this clinical setting is justified. PMID- 15827795 TI - The prognostic effect of clinical staging in pancreatic adenocarcinoma. AB - BACKGROUND: The importance of pancreatic cancer staging is uncertain. The aim of this report was to evaluate the accuracy of combined standard imaging techniques in predicting the pathologic stage and to evaluate the prognostic effect of clinical staging to identify patient groups in which laparoscopy and laparotomy could be beneficial. METHODS: Fifty-four patients were included in this analysis. The techniques used for clinical staging were endoscopic retrograde cholangiopancreatography, abdominal computed tomographic scan, and ultrasonography. All patients underwent both clinical and surgical/pathologic staging. A comparison was performed between presurgical stage and surgical/pathologic stage. The prognostic effect of different factors on survival was evaluated with both univariate (log-rank) and multivariate (Cox) analysis. RESULTS: Sensitivity and specificity for vascular involvement were 73.9% and 96.3%, respectively. Sensitivity and specificity for nodal involvement were 63.6% and 95.4%, respectively. A total of 33.3% of patients showed a higher than expected pathologic stage, and 3.7% showed a lower than expected pathologic stage, by comparing clinical and pathologic evaluation. A highly significant correlation was observed between clinical T stage (P = .0067) and tumor diameter (P = .0037) and patient survival. Maximal prognostic differentiation was observed by dividing patients into two groups based on imaging results: group A (favorable prognosis) and group B (unfavorable prognosis). The median survival was 25.1 and 8.0 months for group A and B, respectively. Five-year survival was 20.1% and 0%, respectively (multivariate analysis: P = .0007). CONCLUSIONS: Integrated standard imaging studies achieved reasonable diagnostic accuracy in our analysis. A single classification based on clinical stage and tumor diameter evaluated by imaging predicts prognosis in patients with pancreatic carcinoma. PMID- 15827796 TI - Electrical impedance scanning of thyroid nodules before thyroid surgery: a prospective study. AB - BACKGROUND: Electrical impedance scanning (EIS) is a novel imaging technique based on differential electrical conductivity and capacitance of malignant and normal human tissues. The aim of this study was to evaluate the accuracy of EIS in the detection of thyroid malignancies. METHODS: Patients with thyroid nodules scheduled for thyroid surgery were eligible for the study. Enrolled patients underwent EIS with a T-Scan 2000ED. Nodule location, size, and type (cystic vs. solid) measured by ultrasound, cytology results, thyroid conductivity, and capacitance calculated by EIS were recorded. EIS results were interpreted as positive or negative for malignancy and compared with final histopathology results. Study end points included EIS accuracy, sensitivity, specificity, negative and positive predictive values, and false-positive and false-negative rates. RESULTS: Sixty-four patients were enrolled onto the study, and all underwent either lobectomy-isthmusectomy (20%) or total thyroidectomy (80%). The mean tumor diameter was 2.64 +/- 14.8 mm. Thyroid cancers were identified by histology in 30 patients (46.9%). There were 11 false-positive and four false negative cases. The overall diagnostic accuracy of EIS was 76.6% (49 of 64 correct diagnoses). The sensitivity and specificity of EIS were 86.7% (26 of 30 true positive) and 67.6% (23 of 34 true negative), respectively. The corresponding positive and negative predictive values were 70.3% and 85.2%. CONCLUSIONS: EIS is a potentially useful imaging modality for differentiating thyroid neoplasms. If these results are confirmed in large-scale trials, EIS may be an important part of the evaluation of thyroid nodules. PMID- 15827797 TI - Immediate reconstruction after neoadjuvant chemotherapy: effect on adjuvant treatment starting and survival. AB - BACKGROUND: For patients treated with initial surgery, the safety of immediate breast reconstruction after mastectomy has been demonstrated. Some concerns exist after neoadjuvant chemotherapy because this sequence is proposed for patients with large tumors and for whom adjuvant therapies are considered cornerstones of treatment. In this study, we sought to determine whether reconstruction after neoadjuvant chemotherapy and mastectomy for large operable breast cancer affects the interval between surgery and adjuvant treatment and affects survival. METHODS: A single-institution retrospective analysis was performed by using the database of the Institut Gustave-Roussy. RESULTS: Forty-eight patients who had undergone mastectomy and immediate reconstruction (implant, 60%) followed by adjuvant chemotherapy were identified. They were compared with 181 patients who underwent mastectomy without reconstruction and with 32 patients who underwent mastectomy followed by delayed reconstruction (implant, 19%). No difference was found concerning the interval between surgery and adjuvant chemotherapy: 26 vs. 23 days for patients with immediate breast reconstruction and for patients treated with modified radical mastectomy followed or not by delayed reconstruction, respectively (P = .11). No difference was found concerning the onset of radiotherapy: 87 vs. 81 days (P = .22). Survival was not different in patients treated with immediate reconstruction compared with those with mastectomy alone. CONCLUSIONS: Immediate breast reconstruction does not delay the starting of adjuvant therapy and has no significant effect on local relapse-free or distant disease-free survival. Additional data are needed concerning the use of flap for this indication. PMID- 15827798 TI - Intraoperative subareolar injection of 99mTc-labeled sulfur colloid results in consistent sentinel lymph node identification. AB - BACKGROUND: Preoperative parenchymal or peritumoral (PT) injection of (99m)Tc labeled sulfur colloid (TcSC) is the standard method for sentinel lymph node (SLN) identification in patients with breast cancer. Limitations of this method include variable identification rates, slow transit times, and painful injections. We hypothesize that TcSC will travel to the SLN within minutes after injection into the subareolar (SA) lymphatics, thus making an intraoperative injection technique feasible. METHODS: One hundred twenty-two women with invasive breast cancer were enrolled onto this prospective study. Immediately after the induction of general anesthesia, patients were injected with 1 to 2 mCi of filtered TcSC in the SA location. Then, 5 mL of 1% isosulfan blue dye was injected into the PT location. The SLN or SLNs were identified as radioactive, blue, or both and removed for pathologic evaluation. RESULTS: The mean patient age was 56 years. The mean tumor size was 1.5 cm. In 86.1% of patients, a transcutaneous axillary "hot spot" was identified by handheld gamma probe. The mean time from TcSC injection to axillary incision was 17.6 minutes. At least one SLN was identified in 99.2% of patients. The mean number of SLNs identified per patient was 1.83. The mean count of radioactive SLNs was 2715 cps. In 97.2% of patients, blue SLNs were also radioactive. CONCLUSIONS: TcSC injected into the SA lymphatics rapidly drains to the SLN. The radioactive SLN is easily and quickly identified after an intraoperative SA TcSC injection. The simplicity of this method eliminates the inherent problems associated with standard PT injection. PMID- 15827799 TI - The results of frozen section, touch preparation, and cytological smear are comparable for intraoperative examination of sentinel lymph nodes: a study in 133 breast cancer patients. AB - BACKGROUND: The goal of intraoperative sentinel lymph node (SLN) examination is to avoid reoperation for a positive SLN, but the ideal method of intraoperative SLN examination remains unclear, and published results vary widely. METHODS: We evaluated the sensitivity of intraoperative frozen section (FS), touch preparation (TP), and cytological smear (CS) in 305 SLNs from 133 breast cancer patients. Each SLN was received fresh and cut into 2- to 3-mm slices; TP and CS from each cut surface and an FS of the entire SLN were obtained. Postoperative evaluation of the SLN consisted of 1 hematoxylin and eosin-stained section and of one hematoxylin and eosin-stained and one immunohistochemically stained section for cytokeratin from each of two levels 50 microm apart. Tumor cells found by any method, including immunohistochemistry, identified a positive SLN. Three pathologists blinded to the final SLN diagnosis reviewed all TP, CS, and FS; the consensus diagnosis (concordance of two or more) was used for the study. RESULTS: FS, TP, and CS had comparable sensitivities (59%, 57%, and 59%, respectively). Each method was more sensitive in detecting macrometastases (>2 mm; 96%, 93%, and 93%, respectively) than micrometastases (< or =2 mm; 27%, 27%, and 30%, respectively). The combination of methods only marginally improved the intraoperative sensitivity. TP and CS were each responsible for a single false positive result. CONCLUSIONS: FS, TP, and CS are comparable for the intraoperative detection of SLN metastases, and each method is substantially better at detecting micrometastases than micrometastases. The combination of two or more techniques only marginally improves the sensitivity over that achieved by a single method. PMID- 15827800 TI - Stereotactic radioguided surgery by siteSelect for subclinical mammographic lesions. AB - BACKGROUND: We defined the indications for and evaluated the results of a new technique for radioguided surgery, the SiteSelect system. The procedure allows en bloc resection of the breast parenchyma under local anesthesia. METHODS: This prospective study was based on 167 patients operated on between December 2000 and October 2003 with 2 phases. The first step was an evaluation of the feasibility of the procedure with the 15-mm cannula, and the second was therapeutic with the 22-mm cannula. RESULTS: The mean duration of the procedure was 42 minutes. In 96.9% of procedures, the lesion was excised successfully. Only one complication (hematoma) and two failures and were observed. Histological examination revealed benign disease in 65.8% of cases and cancer in 34.2% of cases. In the latter cases, the specimen margins were histologically involved in 86.2% of cases with the 15-mm procedure and in 41% with the 22-mm procedure. During the first evaluation, all patients with a cancer underwent systematic surgical re-excision: residual tumor was present in 18 cases (64.2%). The biopsy was painless for 88 patients, and the cosmetic result was good in all cases. CONCLUSIONS: This study shows that the SiteSelect procedure allows resection of the lesion in 96.9% of cases. Combined with complementary surgical lumpectomy during the same operation, this procedure achieved a success rate of 98.7%. In the case of cancer, the 15-mm cannula is not wide enough to allow free margins. The use of a new 22-mm cannula, currently under evaluation, might solve this problem. PMID- 15827801 TI - Popliteal lymph node dissection. AB - Most sentinel nodes are located in the cervical, axillary, and inguinal nodal basins. Sometimes, however, sentinel nodes exist outside these traditional nodal basins. Popliteal nodal metastasis is relatively uncommon, and popliteal lymph node dissection is infrequently necessary. However, with lymphoscintigraphic identification of popliteal sentinel nodes, surgeons are more frequently called on to address the popliteal nodal basin. Therefore, knowledge of the anatomy and surgical technique for popliteal lymphadenectomy is essential. This case study illustrates the importance of considering the approach to the popliteal lymph node basin for patients with melanoma. PMID- 15827803 TI - It is still not the time to change surgical strategy for gastric cancer. PMID- 15827805 TI - 58th Annual Cancer Symposium, Society of Surgical Oncology, March 3-6, 2005, Atlanta, Georgia, USA. Abstracts. PMID- 15827808 TI - Desmoplastic melanoma: a heterogeneous entity in which subclassification as "pure" or "mixed" may have important prognostic significance. PMID- 15827809 TI - Histological and molecular markers for pancreatic adenocarcinoma: can they help us define which patients should receive it? PMID- 15827810 TI - The end of cryotherapy for the treatment of nonresectable hepatic tumors? PMID- 15827811 TI - Ablative therapy for liver cancer: which? PMID- 15827812 TI - Desmoplastic melanoma: a pathologically and clinically distinct form of cutaneous melanoma. AB - BACKGROUND: Desmoplastic melanoma (DM) is a rare variant characterized by the presence of fusiform melanocytes in a sclerotic stroma. Pathologic heterogeneity within DM may account for the controversy regarding the clinical presentation and prognosis of DM compared with conventional melanoma (CM). METHODS: We identified 131 patients with a diagnosis of DM seen between 1979 and 2002. Tumors were categorized as either pure DM (pDM; n = 92), if desmoplasia was prominent throughout the entire invasive tumor, or mixed DM (mDM; n = 39), if fibrosis was well developed in only parts of an otherwise non-DM. Differences in clinical behavior among pDM, mDM, and CM (n = 3976) were examined. RESULTS: Seventy-three percent of patients with DM had tumors >2 mm in depth, compared with 31% of patients with CM (P < .001). Regional nodal metastasis was uncommon in patients who presented with clinically localized pDM (1%) compared with those with mDM (10%) or CM (6%) (P < .05, pDM vs. CM). Five-year melanoma-specific mortality was lower for patients who presented with pDM compared with mDM (11% vs. 31%; P < .01). Patients with pDM and CM had a similar melanoma-specific mortality despite a 3-fold difference in median tumor depth (3.6 vs. 1.2 mm, respectively). CONCLUSIONS: DMs can be divided into two subtypes based on a histological quantification of desmoplasia. Tumors with prominent fibrosis (pure subtype) are unlikely to disseminate to regional lymph nodes and are associated with a favorable outcome when compared with those with mixed desmoplasia or CM. PMID- 15827813 TI - Significance of histological response to preoperative chemoradiotherapy for pancreatic cancer. AB - BACKGROUND: Neoadjuvant (preoperative) chemoradiotherapy (CRT) for pancreatic cancer offers theoretical advantages over the standard approach of surgery followed by adjuvant CRT. We hypothesized that histological responses to CRT would be significant prognostic factors in patients undergoing neoadjuvant CRT followed by resection. METHODS: Since 1994, 193 patients with biopsy-proven pancreatic adenocarcinoma have completed neoadjuvant CRT, and 70 patients have undergone resection. Specimens were retrospectively examined by an individual pathologist for histological responses (tumor necrosis, tumor fibrosis, and residual tumor load) and immunohistochemical staining for p53 and epidermal growth factor receptor. Factors influencing overall survival were analyzed with the Kaplan-Meier (univariate) and Cox proportional hazards (multivariate) methods. RESULTS: The estimated overall survival (median +/- SE) in the entire group of patients undergoing resection was 23 +/- 4.2 months, with an estimated 3 year survival of 37% +/- 6.6% and a median follow-up of 28 months. Complete histological responses occurred in 6% of patients. Overexpression of p53 was more common in patients with large residual tumor loads. Tumor necrosis was an independent negative prognostic factor, as were positive lymph nodes, a large residual tumor load, and poor tumor differentiation. CONCLUSIONS: Histological response to neoadjuvant CRT--as measured by residual tumor load--may be useful as a surrogate marker for treatment efficacy. Characterization of the tumor cells that survive neoadjuvant CRT may help us to identify new or more appropriate targets for systemic therapy. PMID- 15827814 TI - Is pylorospasm a cause of delayed gastric emptying after pylorus-preserving pancreaticoduodenectomy? AB - BACKGROUND: Delayed gastric emptying (DGE) occurs in 14% to 61% of patients after pylorus-preserving pancreaticoduodenectomy, but its pathogenesis is unclear. We hypothesized that DGE may be due to pylorospasm secondary to vagal injuries at operation and may be preventable by the addition of pyloromyotomy. METHODS: Patients operated on consecutively between April 2000 and August 2003 were studied. Pyloromyotomy was of the Fredet-Ramstedt type combined with antroplasty. DGE-free recovery was defined as tolerance of a diet for three successive days by postoperative day 8. The symptom of nausea was used as a basis for nasogastric tube removal and diet resumption. A gastric emptying test (GET) with solid food was obtained. Patients with difficulty swallowing were fed via a feeding tube. RESULTS: There were 47 patients. Two patients were excluded because of death (n = 1) and ileus with pancreatic fistula (n = 1). Diagnoses were pancreatic cancer (n = 23), chronic pancreatitis (n = 11), ampullary cancer (n = 5), mucinous cystic neoplasm (n = 5), and duodenal villous adenoma (n = 3). Median times to nasogastric tube removal, start of liquid diet, and start of solid diet were postoperative days 2, 3, and 5, respectively. Two patients had tube feedings. Preoperative GET was abnormal in 51%, and postoperative GET was abnormal in 37%. The average length of stay was 9.5 days (median, 7 days). DGE occurred in only one patient (2.2%). There were no late complications during a 6-month follow-up. CONCLUSIONS: The addition of pyloromyotomy to pylorus-preserving pancreaticoduodenectomy is effective in preventing DGE. Results are supportive of the hypothesis that DGE may be caused by operative injuries of the vagus innervating the pyloric region. PMID- 15827815 TI - Prognostic factors for survival in patients with locally recurrent extremity soft tissue sarcomas. AB - BACKGROUND: Factors prognostic for survival in patients with locally recurrent extremity soft tissue sarcomas (STS) are poorly defined, thus making it difficult to identify high-risk patients who may benefit from adjuvant therapy. METHODS: A total of 1421 patients underwent surgical treatment for primary extremity STS at a single institution between 1982 and 2002. Of these, 179 (13%) patients underwent complete surgical resection of an isolated local recurrence and were the subjects of this study. Clinicopathologic factors from both the primary tumor and the local recurrence were analyzed. RESULTS: The median interval to initial local recurrence was 16 months: 65% developed a local recurrence by 2 years and 90% by 4 years. Only four patients who presented with a low-grade primary tumor progressed to a high-grade local recurrence. Independent prognostic factors for disease-specific survival after local recurrence were a high histological grade (hazard ratio, 5.1; P = .0004), a large local recurrence tumor size (hazard ratio, 1.5; P = .0001), and a short local recurrence-free interval (hazard ratio, 1.6; P = .0001). Patients who developed a local recurrence >5 cm in < or =16 months (n = 44; 0 low grade) had a 4-year disease-specific survival of 18%, compared with 81% for patients who developed a local recurrence < or =5 cm in >16 months (n = 46; 11 low grade). CONCLUSIONS: Histological grade, local recurrence size, and local recurrence-free interval are independently predictive of survival in patients with locally recurrent extremity STS. A large local recurrence that develops in a short interval indicates a biologically aggressive tumor with a high tumor-specific mortality. Patients who develop such recurrences are ideal subjects for systemic neoadjuvant therapy trials. PMID- 15827816 TI - Radiation-induced sarcoma: a challenge for the surgeon. AB - BACKGROUND: Treatment of radiation-induced sarcoma (RIS) remains an unsolved problem. To provide more insight into the disease process, its characteristics, outcome, and potential outcome determinants were defined. METHODS: From 1978 to 2003, 27 patients--20 females (74%) and 7 males (26%) with a median age 44 years (range, 1-73 years) at the time of diagnosis of the primary tumor--developed an RIS after a median interval of 8 years (range, 3-41 years). The histology of the RIS was 10 (37%) undifferentiated high-grade pleomorphic sarcomas, 7 (26%) angiosarcomas, 6 (22%) fibrosarcomas, 2 (7%) osteosarcomas, 1 (4%) pleomorphic rhabdomyosarcoma, and 1 (4%) pleomorphic leiomyosarcoma. Surgical resection was performed in 21 patients: 13 (62%) R0 (microscopically radical), 4 (19%) R1 (microscopically irradical), 2 (9.5%) R2 (macroscopically irradical), and 2 (9.5%) RX (unknown radicality). Six (22%) patients underwent no resection. RESULTS: The 5-year disease-free and overall survival rates were 27% and 30%, respectively. The local failure rate after R0 resection was 54%. The distant failure rate for the entire group was 41%. Patients with an R0 resection had a significantly better survival rate (P < .05) than patients with an R1, R2, or no resection. CONCLUSIONS: RISs are aggressive malignancies with a high tendency for local recurrence and distant metastases. Previously applied treatment often hampers adequate resection. Therefore, radical surgical resection is the only chance to improve disease-free and overall survival, but it may also have a palliative role. Still, the overall prognosis remains poor. PMID- 15827817 TI - Prognostic value of extracapsular extension of axillary lymph node metastases in T1 to T3 breast cancer. AB - BACKGROUND: The importance of extracapsular extension (ECE) of axillary metastases as a risk factor for either local or distant recurrence and poorer survival in breast cancer has been suggested, but its prognostic value has not been uniformly confirmed. METHODS: From a prospective database including 1142 breast cancer patients operated on at the Department of General Surgery and Surgical Oncology of the University of Siena, we selected 376 cases with pT1 to pT3 node-positive breast cancer. The prognostic significance of ECE of axillary metastases was evaluated with respect to disease-free survival, overall survival, and the patterns of disease recurrence. Such prognostic significance was then compared with that of other clinical and pathologic factors. RESULTS: With a median follow-up of 103 months, factors with independent prognostic value for disease-free survival by multivariate analysis included absence of estrogen receptors (P < .0005), pN category (P < .01), presence of lymphovascular invasion (LVI; P < .005), and ECE (P < .0001). An independent negative prognostic effect on overall survival was observed for absence of estrogen and progesterone receptors (P < .05), pN category (P < .05), and presence of LVI (P < .005) and ECE (P < .0001). The presence of ECE was significantly related to an increased risk of regional (13.4% vs. 6.6%; P = .037) and distant (43% vs. 16.2%; P < .001) recurrences. CONCLUSIONS: ECE demonstrated a stronger statistical significance in predicting prognosis than the pN category and was also related to an increased risk of distant recurrences. We suggest that the decision on adjuvant therapy should consider the presence of ECE of axillary metastases and peritumoral LVI as indicators of high biological aggressiveness. PMID- 15827818 TI - Incidence and clinical significance of lymph node metastasis detected by cytokeratin immunohistochemical staining in ductal carcinoma in situ. AB - BACKGROUND: This study explored the long-term prognosis of patients with ductal carcinoma-in-situ (DCIS) and lymph node metastasis detected by cytokeratin immunohistochemical stains (CK-IHC). METHODS: Using the Columbia University breast cancer database, we identified all DCIS patients who had eight or more axillary nodes dissected and free of metastasis. Five-micrometer sections from all paraffin blocks containing lymph node tissue were stained with an anticytokeratin antibody cocktail (AE1/AE3 and KL1). The results of the CK-IHC and updated database were anonymized and merged. Survival of CK-IHC-positive and negative patients was compared by using Kaplan-Meier curves and log-rank tests. RESULTS: CK-IHC was performed on 301 DCIS patients, who had an average of 16.7 axillary nodes dissected. Eighteen (6%) of 301 patients tested positive by CK IHC. Seventy patients with bilateral breast cancer and 2 patients without any follow-up data were excluded, for a final study population of 229 patients. Among the 216 patients with negative CK-IHC, 18 patients died, compared with 1 of 13 patients with positive CK-IHC. The median follow-up for the study group was 127 months. Kaplan-Meier overall and breast cancer-specific survival estimates were similar for CK-IHC-positive and -negative patients (P = .81 and P = .73, respectively). CONCLUSIONS: CK-IHC increases the incidence of positive nodes by 6% in DCIS patients. A positive node by CK-IHC does not seem to affect survival in these patients. These results raise concerns regarding the clinical significance of positive nodes by CK-IHC in DCIS patients. PMID- 15827819 TI - Is parathyroid carcinoma indeed a lethal disease? AB - BACKGROUND: Parathyroid carcinoma is a rare malignancy with a wide range of aggressiveness. There is no current staging system. Our primary aim was to review the presentation, diagnosis, surgical treatment, and outcomes of patients, with the goal of assessing the incidence of death related to parathyroid carcinoma. METHODS: The authors present a retrospective chart review on patients with parathyroid carcinoma from 1975 to 2004, identified by the tumor registry of a single tertiary-care center. Diagnoses were confirmed histologically, clinical and radiographical data were recorded, and statistical analyses were performed. RESULTS: Twenty-three cases were identified. The mean patient age was 54 years. The female:male ratio was 1.5:1. Follow-up ranged from 1 month to 23 years (median, 134 months). Mean preoperative calcium was 12.9 mg/dL. Median parathyroid hormone was 290 pg/mL. Two patients (9%) had an asymptomatic presentation, and five (22%) presented with a palpable neck mass. Only nine (39%) underwent initial comprehensive en-bloc resection. Median survival was 22 years. Five- and 10-year survival was 85.9% and 69.4%, respectively. Five- and 10-year survival with en-bloc resection was 90% and 67.5%, respectively. Local resection resulted in survival rates of 82.5% and 70.7%. Three of ten deaths were attributed to parathyroid carcinoma. In recurrent disease, computed tomography and scintigraphy had localization rates of 53% and 67%, respectively, with a concordance of 22%. CONCLUSIONS: Long-term survival is possible with parathyroid carcinoma. Death associated with parathyroid carcinoma was uncommon. A staging may be warranted despite the rarity of this disease. PMID- 15827820 TI - Solution for nonuniformities and spatial noise in medical LCD displays by using pixel-based correction. AB - Liquid crystal displays (LCD) are rapidly replacing cathode ray tube displays (CRT) for medical imaging. LCD technology has improved significantly in the last few years and has important advantages over CRT. However, there are still some aspects of LCD that raise questions as to the usefulness of liquid crystal displays for very subtle clinical diagnosis such as mammography. One drawback of modern LCD displays is the existence of spatial noise expressed as measurable stationary differences in the behavior of individual pixels. This type of noise can be described as a random stationary image superposed on top of the medical image being displayed. It is obvious that this noise image can make subtle structures invisible or add nonexistent patterns to the medical image. In the first case, subtle abnormalities in the medical image could remain undetected, whereas in the second case, it could result into a false positive. This paper describes a method to characterize the spatial noise present in high-resolution medical displays and a technique to solve the problem. A medical display with built-in compensation for the spatial noise at pixel level was developed and improved image quality is demonstrated. PMID- 15827821 TI - Functional cluster analysis of CT perfusion maps: a new tool for diagnosis of acute stroke? AB - CT perfusion imaging constitutes an important contribution to the early diagnosis of acute stroke. Cerebral blood flow (CBF), cerebral blood volume (CBV) and time to-peak (TTP) maps are used to estimate the severity of cerebral damage after acute ischemia. We introduce functional cluster analysis as a new tool to evaluate CT perfusion in order to identify normal brain, ischemic tissue and large vessels. CBF, CBV and TTP maps represent the basis for cluster analysis applying a partitioning (k-means) and density-based (density-based spatial clustering of applications with noise, DBSCAN) paradigm. In patients with transient ischemic attack and stroke, cluster analysis identified brain areas with distinct hemodynamic properties (gray and white matter) and segmented territorial ischemia. CBF, CBV and TTP values of each detected cluster were displayed. Our preliminary results indicate that functional cluster analysis of CT perfusion maps may become a helpful tool for the interpretation of perfusion maps and provide a rapid means for the segmentation of ischemic tissue. PMID- 15827822 TI - A clinical evaluation of the image quality computer program, CoCIQ. AB - To provide an objective way of measuring image quality, a computer program was designed that automatically analyzes the test images of a contrast-detail (CD) phantom. The program gives a quantified measurement of image quality by calculating an Image Quality Figure (IQF). The aim of this work was to evaluate the program and adjust it to clinical situations in order to find the detectable level where the program gives a reliable figure of the contrast resolution. The program was applied on a large variety of images with lumbar spine and urographic parameters, from very low to very high image qualities. It was shown that the computer program produces IQFs with small variations and there were a strong linear statistical relation between the computerized evaluation and the evaluation performed by human observers (R2= 0.98). This method offers a fast and easy way of conducting image quality evaluations. PMID- 15827823 TI - Effect of breast density on computer aided detection. AB - PURPOSE: This study was conducted to assess the clinical impact of breast density and density of the lesion's background on the performance of a computer-aided detection (CAD) system in the detection of breast masses (MA) and microcalcifications (MC). MATERIALS AND METHODS: A total of 200 screening mammograms interpreted as BI-RADS 1 and suspicious mammograms of 150 patients having a histologically verified malignancy from 1992 to 2000 were selected by using a sampler of tumor cases. Excluding those cases having more than one lesion or a contralateral malignancy attributable to statistical reasons, 127 cases with 127 malignant findings were analyzed with a CAD system (Second Look 5.0, CADx Systems, Inc., Beavercreek, OH). Of the 127 malignant lesions, 56 presented as MC and 101 presented as MA, including 30 cases with both malignant signs. Overall breast density of the mammogram and density of the lesion's background were determined by two observers in congruence (density a: entirely fatty, density b: scattered fibroglandular tissue, density c: heterogeneously dense, density d: extremely dense). RESULTS: Within the unsuspicious group, 100/200 cases did not have any CAD MA marks and were therefore truly negative (specificity 50%), and 151/200 cases did not have any CAD MC marks (specificity 75.5%). For these 200 cases, the numbers of marks per image were 0.41 and 0.37 (density a), 0.38 and 0.97 (density b), 0.44 and 0.91 (density c), and 0.58 and 0.68 (density d) for MC and MA marks, respectively (Fisher's t-test: n.s. for MC, p < 0.05 for MA). Malignant lesions were correctly detected in at least one view by the CAD system for 52/56 (92.8%) MC and 91/101 (90.1%) MA. Detection rate versus breast density was: 4/6 (66.7%) and 18/19 (94.7%) (density a), 32/33 (97.0%) and 49/51 (96.1%) (density b), 14/15 (93.3%) and 23/28 (82.1%) (density c), and 2/2 (100%) and 1/3 (33.3%) (density d) for MC and MA, respectively. Detection rate versus the lesion's background was: 19/21 (90.5%) and 36/38 (94.7%) (density a), 34/36 (94.4%) and 59/62 (95.2%) (density b), 8/9 (88.9%) and 20/24 (83.3%) (density c), and 9/10 (90%) and 4/8 (50%) (density d) for groups 2 and 3, respectively. Detection rates differed significantly for masses in heterogeneously dense and extremely dense tissue (overall or lesion's background) versus all other densities (Fisher's t-test: p < 0.05). A significantly lowered FP rate for masses was found on mammograms of entirely fatty tissue. CONCLUSION: Overall breast density and density at a lesion's background do not appear to have a significant effect on CAD sensitivity or specificity for MC. CAD sensitivity for MA may be lowered in cases with heterogeneously and extremely dense breasts, and CAD specificity for MA is highest in cases with extremely fatty breasts. The effects of overall breast density and density of a lesion's background appear to be similar. PMID- 15827824 TI - Computerized methods for determining respiratory phase on dynamic chest radiographs obtained by a dynamic flat-panel detector (FPD) system. AB - Chest radiography using a dynamic flat-panel detector with a large field of view can provide sequential chest radiographs during respiration. These images provide information regarding respiratory kinetics, which is effective for diagnosis of pulmonary diseases. For valid analysis of respiratory kinetics in diagnosis of pulmonary diseases, it is crucial to determine the association between the kinetics and respiratory phase. We developed four methods to determine the respiratory phase based on image information associated with respiration and compared the results in dynamic chest radiographs of 37 subjects. Here, the properties of each method and future tasks are discussed. The method based on the change in size of the lung gave the most stable results, and that based on the change in distance from the lung apex to the diaphragm was the most promising method for determining the respiratory phase. PMID- 15827825 TI - Evaluating different radiology workstation interaction techniques with radiologists and laypersons. AB - This paper presents a new methodology for evaluating radiology workstation interaction features, using lay subjects to perform a radiology look-alike task with artificial stimuli. We validated this methodology by evaluating two different workstation interaction techniques with two groups of subjects: laypersons and radiologists, using a set of artificial targets to simulate the reading of a diagnostic examination. Overall, the results from the two groups of subjects performing the same tasks were very similar. Both groups showed significantly faster response times using a new interaction technique, and the mouse clicks for both groups were very similar, showing that all the subjects mastered the style of interaction in a similar way. The errors made by both groups were comparable. These results show that it is possible to test new workstation interaction features using look-alike radiological tasks and inexperienced laypersons, and that the results do transfer to radiologists performing the same tasks. PMID- 15827826 TI - Collaborative filtering to improve navigation of large radiology knowledge resources. AB - OBJECTIVE: Collaborative filtering is a knowledge-discovery technique that can help guide readers to items of potential interest based on the experience of prior users. This study sought to determine the impact of collaborative filtering on navigation of a large, Web-based radiology knowledge resource. MATERIALS AND METHODS: Collaborative filtering was applied to a collection of 1,168 radiology hypertext documents available via the Internet. An item-based collaborative filtering algorithm identified each document's six most closely related documents based on 248,304 page views in an 18-day period. Documents were amended to include links to their related documents, and use was analyzed over the next 5 days. RESULTS: The mean number of documents viewed per visit increased from 1.57 to 1.74 (P < 0.0001). CONCLUSIONS: Collaborative filtering can increase a radiology information resource's utilization and can improve its usefulness and ease of navigation. The technique holds promise for improving navigation of large Internet-based radiology knowledge resources. PMID- 15827827 TI - Assessment of real-time 3D visualization for cardiothoracic diagnostic evaluation and surgery planning. AB - RATIONALE AND OBJECTIVES: Three-dimensional (3D) real-time volume rendering has demonstrated improvements in clinical care for several areas of radiological imaging. We test whether advanced real-time rendering techniques combined with an effective user interface will allow radiologists and surgeons to improve their performance for cardiothoracic surgery planning and diagnostic evaluation. MATERIAL AND METHODS: An interactive combination 3D and 2D visualization system developed at the University of North Carolina at Chapel Hill was compared against standard tiled 2D slice presentation on a viewbox. The system was evaluated for 23 complex cardiothoracic computed tomographic (CT) cases including heart-lung and lung transplantation, tumor resection, airway stent placement, repair of congenital heart defects, aortic aneurysm repair, and resection of pulmonary arteriovenous malformation. Radiologists and surgeons recorded their impressions with and without the use of the interactive visualization system. RESULTS: The cardiothoracic surgeons reported positive benefits to using the 3D visualizations. The addition of the 3D visualization changed the surgical plan (65% of cases), increased the surgeon's confidence (on average 40% per case), and correlated well with the anatomy found at surgery (95% of cases). The radiologists reported fewer and less major changes than the surgeons in their understanding of the case due to the 3D visualization. They found new findings or additional information about existing findings in 66% of the cases; however, they changed their radiology report in only 14% of the cases. CONCLUSION: With the appropriate choice of 3D real-time volume rendering and a well-designed user interface, both surgeons and radiologists benefit from viewing an interactive 3D visualization in addition to 2D images for surgery planning and diagnostic evaluation of complex cardiothoracic cases. This study finds that 3D visualization is especially helpful to the surgeon in understanding the case, and in communicating and planning the surgery. These results suggest that including real-time 3D visualization would be of clinical benefit for complex cardiothoracic CT cases. PMID- 15827828 TI - Digital radiographic image denoising via wavelet-based hidden Markov model estimation. AB - This paper presents a technique for denoising digital radiographic images based upon the wavelet-domain Hidden Markov tree (HMT) model. The method uses the Anscombe's transformation to adjust the original image, corrupted by Poisson noise, to a Gaussian noise model. The image is then decomposed in different subbands of frequency and orientation responses using the dual-tree complex wavelet transform, and the HMT is used to model the marginal distribution of the wavelet coefficients. Two different correction functions were used to shrink the wavelet coefficients. Finally, the modified wavelet coefficients are transformed back into the original domain to get the denoised image. Fifteen radiographic images of extremities along with images of a hand, a line-pair, and contrast detail phantoms were analyzed. Quantitative and qualitative assessment showed that the proposed algorithm outperforms the traditional Gaussian filter in terms of noise reduction, quality of details, and bone sharpness. In some images, the proposed algorithm introduced some undesirable artifacts near the edges. PMID- 15827829 TI - A novel application of the MIRC repository in medical education. AB - Medical students on the radiology elective in our institution create electronic presentations to present to each other as part of the requirements for the rotation. Access was given to previous students' presentations via the web-based system, Medical Imaging Resource Center (MIRC) project, created and supported by the Radiological Society of North America (RSNA). RadPix Power 2 MIRC (Weadock Software, LLC, Ann Arbor, MI) software converted the Microsoft PowerPoint (Redmond, WA) presentations to a MIRC-compatible format. The textual information on each slide is searchable across the entire MIRC database. Future students will be able to benefit from the work of their predecessors. PMID- 15827830 TI - Designing better radiology workstations: impact of two user interfaces on interpretation errors and user satisfaction. AB - This paper presents our solution for supporting radiologists' interpretation of digital images by automating image presentation during sequential interpretation steps. We extended current hanging protocols with support for "stages" which reflect the presentation of digital information required to complete a single step within a complex task. We demonstrated the benefits of staging in a user experiment with 20 lay subjects involved in a comparative visual search for targets, similar to a radiology task of identifying anatomical abnormalities. We designed a task and a set of stimuli that allowed us to simulate the interpretation workflow from a typical radiology scenario-reading a chest radiography exam when a prior study is also available. The simulation was enabled by abstracting both the radiologist's task and the basic workstation navigation functionality. The staged interface was significantly faster than the traditional user interface, provided a 37% reduction in the interpretation errors, and improved user satisfaction. PMID- 15827831 TI - Creation of DICOM--aware applications using ImageJ. AB - The demand for image-processing software for radiology applications has been increasing, fueled by advancements in both image-acquisition and image-analysis techniques. The utility of existing image-processing software is often limited by cost, lack of flexibility, and/or specific hardware requirements. In particular, many existing packages cannot directly utilize images formatted using the specifications in part 10 of the DICOM standard ("DICOM images"). We show how image analyses can be performed directly on DICOM images by using ImageJ, a free, Java-based image-processing package (http://rsb.info.nih.gov/ij/). We demonstrate how plug-ins written in our laboratory can be used along with the ImageJ macro script language to create flexible, low-cost, multiplatform image-processing applications that can be directed by information contained in the DICOM image header. PMID- 15827832 TI - Impact of PACS and voice-recognition reporting on the education of radiology residents. AB - RATIONALE AND OBJECTIVES: The introduction of picture archiving and communication system (PACS) has decreased the time needed to interpret radiology examinations resulting in an increased workflow. Because of concerns that the increase in exam throughput and the use of voice recognition may have a negative impact upon radiology resident education, a survey was conducted to assess the impact of PACS and voice recognition. MATERIALS AND METHODS: Residents at four diagnostic radiology training programs were surveyed. Survey topics included resident demographics, didactic and technical issues, and areas for improvement. RESULTS: One hundred thirty-four residents were polled with 42 respondents (42/134, 31.3%). The majority have been using PACS for more than 1 year (29/41, 70.7%) to interpret 75-100% of cases (33/39, 84.6%). A majority believed PACS is a superior teaching tool to printed film (28/38, 73.7%). However, only a minority (9/40, 22.5%) indicated that PACS was always used to contain teaching files and to conduct departmental conferences (5/40, 12.5%). The majority of respondents believed PACS have decreased the time needed to interpret diagnostic examinations (33/41, 80.5%). A majority (80.6%, 25/31) indicated that voice recognition takes more time than the traditional dictation and transcription process, where 51.3% (20/39, 51.3%) felt that voice recognition works well less than 50% of the time. CONCLUSIONS: Residents believe that PACS has positively affected their learning experience but indicate that it can be better utilized for resident education. Residents believe that voice recognition is less reliable and more time consuming than the traditional dictation system. PMID- 15827833 TI - Effects of dissolved and complexed copper on heterotrophic bacterial production in San Diego bay. AB - Bacterial abundance and production, free (uncomplexed) copper ion concentration, total dissolved copper concentration, dissolved organic carbon (DOC), total suspended solids (TSS), and chlorophyll a were measured over the course of 1 year in a series of 27 sample "Boxes" established within San Diego Bay. Water was collected through a trace metal-clean system so that each Box's sample was a composite of all the surface water in that Box. Bacterial production, chlorophyll a, TSS, DOC, and dissolved copper all generally increased from Box 1 at the mouth of the Bay to Box 27 in the South or back Bay. Free copper ion concentration generally decreased from Box 1 to Box 27 presumably due to increasing complexation capacity within natural waters. Based on correlations between TSS, chlorophyll a, bacterial production or DOC and the ratio of dissolved to free Cu ion, both DOC and particulate (bacteria and algae) fractions were potentially responsible for copper complexation, each at different times of the year. CuCl2 was added to bacterial production assays from 0 to 10 microg L(-1) to assess acute copper toxicity to the natural microbial assemblage. Interestingly, copper toxicity appeared to increase with decreases in free copper from the mouth of the Bay to the back Bay. This contrasts the free-ion activity model in which higher complexation capacity should afford greater copper protection. When cell-specific growth rates were calculated, faster growing bacteria (i.e. toward the back Bay) appeared to be more susceptible to free copper toxicity. The protecting effect of natural dissolved organic material (DOM) concentrated by tangential flow ultrafiltration (>1 kDa), illite and kaolinite minerals, and glutathione (a metal chelator excreted by algae under copper stress) was assessed in bacterial production assays. Only DOM concentrate offered any significant protection to bacterial production under increased copper concentrations. Although the potential copper protecting agents were allowed to interact with added copper before natural bacteria were added to production assays, there may be a temporal dose-response relationship that accounts for higher toxicity in short production assays. Regardless, it appears that effective natural complexation of copper in the back portions of San Diego Bay limits exposure of native bacterial assemblages to free copper ion, resulting in higher bacterial production. PMID- 15827834 TI - Malignant triton tumor presenting as a rectal mass in an 11-month-old. AB - We report the case of an 11-month-old white male who had a family history of neurofibromatosis, had multiple cafe-au-lait spots on the trunk and extremities, and was diagnosed with a malignant triton tumor of the rectum. To our knowledge, this is the first report of a malignant triton tumor of the rectum and one of the youngest patients reported with the tumor. PMID- 15827836 TI - Patient's preference and randomization: new paradigm of evidence-based clinical research. AB - The limitations associated with the traditional randomized controlled design as applied to clinical surgical research must be recognized. The aim of randomization is to ensure initial comparability between groups of eligible patients for whom treatments are compared, thus eliminating their individual influence on outcome. Randomized controlled trials in the surgical literature are sparse; patient preferences might be a major obstacle to their performance. External validity of results of clinical trials depends on the representativity of patients who participate in trials: Compliance to participate through informed consent may act as a selection bias. In surgical randomized trials where it is not often possible for patients to remain blinded to the treatment to which they have been allocated, patient preferences can influence the effectiveness of treatments. In this setting, we need to look at alternatives and the potential advantages of adopting more flexible and clinically relevant approaches to the design of surgical trials. We have to accept the weight of the patient's individual decision in everyday practice. Hence, to negate the importance of these individual choices when evaluating surgical outcomes is unrealistic. An original design reported herein might become a new paradigm for surgical evaluation. PMID- 15827835 TI - Limits of evidence-based surgery. AB - Evidence-based medicine can be summarized as the use of current best evidence in the care of individual patients. When applied to surgical practice, it appears clearly that the concept of evidence-based medicine involves some limitations. To discuss these limits, the author went back over the terms of the evidence-based medicine definition. Limits are related to the low quantity and quality of randomized controlled trials and meta-analyses in surgery, the difficulties when critically appraising the literature and applying the results of evidence to individual patients, and bringing surgeons more willing to endorse the principles of evidence-based medicine. However all these limits can be overcome in the future, with the aim that evidence-based surgery will not be a passing fad. PMID- 15827837 TI - How to teach evidence-based surgery. AB - The objectives of teaching evidence-based surgery (EBS) are to inform and convince that EBS is a method of interrogation, reasoning, appraisal, and application of information to guide physicians in their decisions to best treat their patients. Asking the right, answerable questions, translating them into effective searches for the best evidence, critically appraising evidence for its validity and importance, and then integrating EBS with their patients' values and preferences are daily chores for all surgeons. Teaching and learning EBS should be patient-centered, learner-centered, and active and interactive. The teacher should be a model for students to become an expert clinician who is able to match and take advantage of the clinical setting and circumstances to ask and to answer appropriate questions. The process is multistaged. Teaching EBS in small groups is ideal. However, it is time-consuming for the faculty and must be clearly and formally structured. As well, evidence-based medicine (EBM) courses must cater to local institutional needs, must receive broad support from the instructors and the providers of information (librarians and computer science faculty), use proven methodologies, and avoid scheduling conflicts. In agreement with others, we believe that the ideal moment to introduce the concepts of EBM into the curriculum of the medical student is early, during the first years of medical school. Afterward, it should be continued every year. When this is not the case, as in many countries, it becomes the province of the surgeon in teaching hospitals, whether they are at the university, are university-affiliated, or not, to fulfill this role. PMID- 15827838 TI - Evidence-based medicine in surgical decision making. AB - There are now five classic steps for analysis of diagnostic and therapeutic medical decision-making policies: (1) formulate a clear clinical question based on a particular patient's problem; (2) search the literature for relevant clinical articles; (3) evaluate the evidence for its validity and usefulness; (4) implement useful findings into clinical practice; (5) audit the validity of the process. The clinician must have the necessary skills to appraise critically the information retrieved. Rather than focusing on the discussion and conclusion sections of articles, the reader should concentrate on the review of the methods and results sections to formulate an opinion regarding the strength of evidence presented in the paper. The process is intellectually demanding and difficult to achieve. This particular step in the validation of evidence implies that each clinician must be methodologically and statistically sound, an "expert," capable of analyzing the method used in that particular publication to achieve the published result. PMID- 15827839 TI - Role of systematic reviews and meta-analysis in evidence-based medicine. AB - The overwhelming increase in the quantity of clinical evidence has led to detachment of the evidence and practice because new evidence can be integrated into clinical practice only after it has been critically appraised and synthesized on the basis of the existing evidence. Because many clinicians lack the skills and the time for such information processing, systematic reviews and meta-analyses, their quantitative counterparts, play an important role in health care. Well performed systematic reviews provide clinically relevant information for surgeons, abrogating the need to identify, read, and evaluate many individual studies. This article reviews the basic principles of meta-analysis, discusses its potential weaknesses such as heterogeneity and publication bias, and highlights special situations when dealing with surgical trials. PMID- 15827840 TI - Number needed to treat (or harm). AB - The effect of a treatment versus controls may be expressed in relative or absolute terms. For rational decision-making, absolute measures are more meaningful. The number needed to treat, the reciprocal of the absolute risk reduction, is a powerful estimate of the effect of a treatment. It is particularly useful because it takes into account the underlying risk (what would happen without the intervention?). The number needed to treat tells us not only whether a treatment works but how well it works. Thus, it informs health care professionals about the effort needed to achieve a particular outcome. A number needed to treat should be accompanied by information about the experimental intervention, the control intervention against which the experimental intervention has been tested, the length of the observation period, the underlying risk of the study population, and an exact definition of the endpoint. A 95% confidence interval around the point estimate should be calculated. An isolated number needed to treat is rarely appropriate to summarize the usefulness of an intervention; multiple numbers needed to treat for benefit and harm are more helpful. Absolute risk reduction and number needed to treat should become standard summary estimates in randomized controlled trials. PMID- 15827842 TI - How to appraise a diagnostic test. AB - Clinicians frequently confront challenges when using diagnostic tests to help them decide whether the patient before them suffers from a particular target condition or diagnosis. The primary issues to consider when determining the validity of a diagnostic test study are how the authors assembled the patients and whether they used an appropriate reference standard in all patients to determine whether the patients did or did not have the target condition. Surgeons should be interested in the characteristics of the test that indicates the direction and magnitude of change in the probability of the target condition associated with a particular test result. The likelihood ratio best captures the link between the pretest probability of the target condition and the probability after the test results are obtained (also called the posttest probability). Many studies, however, present the properties of diagnostic tests in less clinically useful terms: sensitivity and specificity. Sensitivity denotes the proportion of people with the disorder in whom the test result is positive. Specificity denotes the proportion of people without the disorder in whom the test result is negative. Application of the guides presented in this article can allow surgeons to assess critically studies regarding a diagnostic test. PMID- 15827841 TI - How to appraise the effectiveness of treatment. AB - Before implementing a new therapy, we should ascertain the benefits and risks of the therapy and assure ourselves that the resources consumed during the intervention will not be exorbitant. In the hierarchy of research designs, the results of randomized controlled trials, especially if systematically reviewed, are considered the highest level of evidence. We suggest a three-step approach to using an article from the medical literature to guide your patient care. We recommend that readers ask whether the study can provide valid results, review the results, and consider how the results can be applied to patient care. Given the time constraints of busy surgical practices and surgical training programs, applying this analysis to every relevant article will be challenging. However, the basis of this process is essentially what we all do many times each week when making decisions about whether and how to treat patients. Making this process explicit with guidelines to assess the strength of the available evidence can serve to improve patient care. It also allow us to defend therapeutic interventions based on available evidence and not anecdote. PMID- 15827843 TI - How to analyze an article. AB - In clinical research investigators generalize from study samples to populations, and in evidence-based medicine practitioners apply population-level evidence to individual patients. The validity of these processes is assessed through critical appraisal of published articles. Critical appraisal is therefore a core component of evidence-based medicine (EBM). The purpose of critical appraisal is not one of criticizing for criticism's sake. Instead, it is an exercise in assigning a value to an article. A checklist approach to article appraisal is outlined, and common pitfalls of analysis are highlighted. Relevant questions are posed for each section of an article (introduction, methods, results, discussion). The approach is applicable to most clinical surgical research articles, even those of a nonrandomized nature. Issues specific to evidence-based surgical practice, in contrast to evidence-based medicine, are introduced. PMID- 15827844 TI - Why we need confidence intervals. AB - The estimation approach to statistical analysis aims to quantify the effect of interest as an "estimate" of a clinically relevant quantity and to quantify the uncertainty in this estimate by means of a confidence interval (CI). As such, results expressed in this form are much more informative than results presented just as p values. This article focuses on the principles rather than the mathematics of CIs and discusses interpretation of CIs and some common misuses. CIs can be constructed for almost all analyses. They are especially useful for avoiding misinterpretation of nonsignificant results of small studies. CIs should be provided routinely for the main results of trials and observational studies. PMID- 15827845 TI - History and development of evidence-based medicine. AB - This article illustrates the timeline of the development of evidence-based medicine (EBM). The term "evidence-based medicine" is relatively new. In fact, as far as we can tell, investigators from McMaster's University began using the term during the 1990s. EBM was defined as "a systemic approach to analyze published research as the basis of clinical decision making." Then in 1996, the term was more formally defined by Sacket et al., who stated that EBM was "the conscientious and judicious use of current best evidence from clinical care research in the management of individual patients." Ancient era EBM consists of ancient historical or anecdotal accounts of what may be loosely termed EBM. This was followed by the development of the renaissance era of EBM, which began roughly during the seventeenth century. During this era personal journals were kept and textbooks began to become more prominent. This was followed by the 1900s, during an era we term the transitional era of EBM (1900-1970s). Knowledge during this era could be shared more easily in textbooks and eventually peer reviewed journals. Finally, during the 1970s we enter the modern era of EBM. Technology has had a large role in the advancement of EBM. Computers and database software have allowed compilation of large amounts of data. The Index Medicus has become a medical dinosaur of the past that students of today likely do not recognize. The Internet has also allowed incredible access to masses of data and information. However, we must be careful with an overabundance of "unfiltered" data. As history, as clearly shown us, evidence and data do not immediately translate into evidence based practice. PMID- 15827846 TI - What should surgeons know about evidence-based surgery. PMID- 15827847 TI - Risk factors for sepsis and endocarditis and long-term survival following coronary artery bypass grafting. AB - We sought to determine risk factors for sepsis and/or endocarditis (S/E) and to identify their impact on long-term survival after coronary artery bypass grafting (CABG). We studied 3760 consecutive patients who underwent isolated CABG from 1992 to 2002. Patients with CABG without S/E were compared with those who developed S/E. Long-term survival data (mean follow-up 5.2 years) were obtained from the National Death Index. Groups were compared by Cox proportional hazard models and Kaplan-Meier survival plots. The propensity for S/E was determined by logistic regression analysis and each patient with S/E was matched to one patient without S/E. Thirty-six patients (0.96%) developed S/E. Independent predictors for S/E were increased age (odds ratio [OR] 1.05 per year, 95% Confidence interval [95% CI] 1.00-1.09; p = 0.040) and the development of other major complications after CABG such as deep sternal wound infection (OR 30.80, 95% CI 9.50-99.82; p < 0.001), gastrointestinal complications (OR 19.48, 95% CI 7.14 53.18; p < 0.001), renal failure (OR 15.18, 95% CI 4.42-52.06; p < 0.001), intraoperative stroke (OR 13.11, 95% CI 4.81-35.69; p < 0.001) and respiratory failure (OR 12.95, 95% CI 5.69-29.45; p < 0.001). After adjustment for pre-, intra- and postoperative factors, the adjusted hazard ratio of long-term mortality for patients with S/E was 3.33 (95% CI 2.17-5.10; p < 0.001). There was no difference in 30-day mortality between matched groups (25.0% vs. 19.4% in patients without S/E, p = 0.778), however patients without S/E had better 5-year survival rate (52.7 +/- 8.7% vs. 16.2 +/- 6.2%; p = 0.0004). We have identified risk factors for S/E following CABG and we found that there was increased mortality in patients with S/E during a 10-year follow-up period. PMID- 15827848 TI - Parathyroid autotransplantation during total thyroidectomy--does the number of glands transplanted affect outcome? AB - Parathyroid autotransplantation is a technique for ensuring the continued function of parathyroid tissue at the time of total thyroidectomy (TT). The aim of this study was to ascertain whether the number of parathyroids transplanted affects the incidence of temporary and permanent hypoparathyroidism. A retrospective cohort study included all patients undergoing a TT in a single unit between July 1998 and June 2003. The number of parathyroids transplanted, the final pathology, and the incidence of temporary and permanent hypoparathyroidism were documented. Fisher's exact test was used for statistical analysis. A total of 1196 patients underwent a TT during the 5 years studied. Of these, 306 (25.6%) had no parathyroids transplanted, 650 (54.3%), 206 (17.2%), 34 (2.8%) had 1,2, or 3 glands autotransplanted, respectively. The incidence of temporary hypoparathyroidism was 9.8% for no gland transplants, 11.9%, 15.1%, and 31.4% for 1,2,and 3 gland transplants, respectively (p < 0.05). The incidence of permanent hypoparathyroidism was 0.98%, 0.77%, 0.97%, and 0%, respectively (p = NS). The incidence of temporary hypoparathyroidism was higher when surgery was performed for Graves' disease. Temporary hypocalcemia is closely related to the number of autotransplanted parathyroids during TT. The long-term outcome is not affected by the number of parathyroids autotransplanted. A "ready selective" approach to parathyroid autotransplantation is an effective strategy for minimizing the rate of permanent hypoparathyroidism. PMID- 15827849 TI - Management of ascites after radical surgery in gastric cancer patients with liver cirrhosis and minimal hepatic dysfunction. AB - A radical lymph node dissection is important for the cure of gastric cancer. However, such a procedure in patients with liver cirrhosis (LC) could develop serious complications such as massive ascites. To determine the management of postoperative ascites, 26 gastric cancer patients with LC were reviewed retrospectively. Child-Pugh status was grade A in all 26 patients. Thirteen (50%) patients had advanced gastric cancer, and a D2 lymph nodes dissection was performed in 25 (96.2%) patients. The mean number of dissected lymph nodes was 33 +/- 11 (range: 11-54). An abdominal closed suction drain was placed in 12 (46.2%) patients, and the average amount of fluid drainage was 463 ml/day. The drainage tube was removed on about the eleventh postoperative day (range: day 6 to day 13), and diuretics were used in 8 (30.8%) patients. A paracentesis was needed in one patient but no postoperative surgical morbidity or mortality was observed. Therefore, an extended lymph node dissection is safe in gastric cancer patients with mild hepatic dysfunction. Liver cirrhosis and postoperative ascites can be managed conservatively without any complications. PMID- 15827850 TI - Hemodynamic and respiratory effects of robot-assisted laparoscopic fundoplication in children. AB - Laparoscopic fundoplication is increasingly used for treating gastro-esophageal reflux disease in children. Mechanical and pharmacological effects may contribute to hemodynamic and respiratory changes during carbon dioxide pneumoperitoneum. The aim of the present study was to evaluate the hemodynamic and respiratory effects of pneumoperitoneum (PP) with an intra-abdominal pressure (IAP) of 12 mmHg in children undergoing robot-assisted laparoscopic fundoplication during total intravenous anesthesia. Ten children, aged 8-16 years, American Society of Anesthesiologists physical status II-III, scheduled for robot-assisted laparoscopic fundoplication in the reverse Trendelenburg position were investigated. Minute ventilation (MV), peak inspiratory pressure (PIP), IAP, heart rate (HR), mean arterial blood pressure (MAP) were recorded, together with pH, base excess, HCO3-, P(et)CO2, PaCO2, and PaO2 at six time points: before insufflation, 10, 30, 60, 90 minutes after creating PP and after desufflation. The IAP was maintained at 12 mmHg. During insufflation MAP increased significantly from 70.6 (+/-9.0) to 84.8 (+/-10.4) mmHg, MV was increased from 4.6 (+/-0.8) to 5.5 (+/-0.9) l min(-1), PIP increased, PaO2 and pH decreased. P(et)CO2 increased from 33.1 (+/-1.6) to 36.6 (+/-1.6) mmHg together with PaCO2. Hemodynamic and respiratory effects due to the intra-abdominal insufflation of CO2 with an IAP of 12 mmHg are well tolerated, and anesthesia with remifentanil, propofol and mivacurium facilitates extubation immediately at the end of surgery. PMID- 15827853 TI - Canthus. PMID- 15827854 TI - The ethics of placebo-controlled trials: a comparison of inert and active placebo controls. AB - Because of the recent and controversial example of sham surgery for the evaluation of fetal tissue transplants for Parkinson's disease, there is renewed interest in the ethics of using "active" placebos in surgical trials, where otherwise there are no inert procedures available, and in pharmacological trials, where there are inert substances, but where patients may guess to which arm they have been allocated. This paper seeks to clarify the ethical arguments surrounding the use of active placebos in trials, and to set up a notation for assessing the ethics of trials more generally. We first establish an framework by which ethics committees can analyze such trials. We examine (1) the scientific value of the research; (2) the expected risks and benefits to individual patients, and (3) the voluntary nature of consent. We then contrast the implications of this framework for inert and active placebo-controlled trials, respectively. In particular, we analyze their relative expected utility using three main utility factors, namely, treatment effects, placebo effects, and altruism. We conclude that, when the intervention is already widely available, active placebo trials rely more heavily on altruism than do inert placebo trials and, when the intervention is restricted, this excess reliance may not be needed. What our analysis provides is the explicit justification for the apparent caution of Institutional Review Boards or ethics committees when reviewing sham operations, especially when the expected harm is not trivial and the risk of exploitation is high. PMID- 15827855 TI - Transduodenal resection of peri-ampullary lesions. AB - Transduodenal resection (TDR) of lesions near the ampulla of Vater is an alternative to the Whipple pancreaticoduodenectomy. A retrospective analysis was performed to determine the long-term outcome and the utility of intraoperative frozen section examinations in aiding operative decision making in patients undergoing TDR. From 1992 to 2002, 19 patients with an average age of 64.2 years (range: 33-84 years) underwent a transduodenal resection of a peri-ampullary lesion; median follow-up was 47 months (range: 2-100 months). Pathology of the lesions was as follows: 11 with benign ampullary adenomas, including 4 with familial adenomatous polyposis (FAP); 7 with peri-ampullary adenocarcinomas; and 1 with a benign stricture. Survival for the entire cohort is 100%. In 12 cases an intraoperative frozen section was performed. The specificity and positive predictive value of the intraoperative histology were both 100%, and the sensitivity and negative predictive value were 57% and 38%, respectively. Three of the 4 patients with FAP have recurrent adenomatous change; 2 of the 7 with carcinoma have metastatic adenocarcinoma. Transduodenal resection of peri ampullary lesions appears to be a safe alternative to radical resection for benign adenomas and selected carcinoma. Intraoperative frozen section assessment is recommended in cases of potential adenocarcinoma. PMID- 15827856 TI - Local platelet trapping as the cause of thrombocytopenia after hepatic cryotherapy. AB - Cryosurgery has been shown to be an effective approach to destruction of unresectable hepatic tumors. However, hepatic cryoablation may also be associated with local and systemic side effects, including thrombocytopenia and clotting dysfunction. Although thrombocytopenia is known to relate to the magnitude of hepatocellular injury, its etiology is still unknown. With the use of whole-body scintigraphy after injection of indium-111-labeled platelets we here demonstrated in six patients undergoing cryoablation of hepatic tumors that manifestation of systemic thrombocytopenia after cryosurgery is associated with excessive platelet trapping and destruction within the cryolesion. We therefore conclude that local platelet trapping represents a major cause of cryothermia-induced systemic thrombocytopenia. PMID- 15827857 TI - Calciphylaxis: a rare complication of patients who required parathyroidectomy for advanced renal hyperparathyroidism. AB - Calciphylaxis is a relatively rare but life-threatening complication in uremic patients. Clinical findings and prognosis were evaluated in six patients who developed calciphylaxis from a group of 1499 patients who underwent parathyroidectomy (PTx) for advanced renal hyperparathyroidism (HPT) in our department from July 1972 to July 2003. The frequency of calciphylaxis was 0.40% (6/1499). Two patients were women and four were men. The mean age was 50.5 years, and the mean duration of hemodialysis (HD) treatment was 14.0 years. In five of six patients, calciphylaxis was classified as distal type; in one case, as proximal type. In three patients, calciphylaxis was diagnosed at the time for PTx. In two patients, calciphylaxis was identified after PTx, although the serum parathyroid hormone (PTH) level was within the appropriate range for dialysis patients. In two patients, calciphylaxis improved after PTx, but two patients required leg and toe amputations after PTx. In one patient with the proximal type of calciphylaxis, the condition occurred when a high PTH level recurred after the initial PTx. The patient died as a result of a serious infection due to uncontrollable skin ulcers. Calciphylaxis is a rare complication in patients who require PTx for renal HPT. Especially the proximal type has a poor prognosis. High levels of the Ca x P product and/or PTH are risk factors. Therefore, this syndrome should be kept in mind and attention should be paid to reduce risk factors. It is important that PTx being performed at the right time in patients with advanced renal HPT refractory to medical treatment. PMID- 15827858 TI - Laparoscopic cardial calibration and gastropexy for treatment of patients with reflux esophagitis: pathophysiological basis and result. AB - Laparoscopic antireflux surgery is the gold standard procedure for treatment of patients with reflux esophagitis. The current results of the laparoscopic approach are absolutely comparables with the results obtained during the open surgery era. The Nissen, Nissen-Rossetti, or Toupet techniques are the more frequently used. We have performed cardial calibration and posterior gastropexy or Nissen fundoplication by the open approach with similar results. The purpose of this article is to present the anatomo-physiological basis for employing cardial calibration and posterior gastropexy in patients with reflux esophagitis. This study includes 108 symptomatic patients, 12 of them with associated extraesophageal manifestations ( posterior laryngitis). Endoscopic mild or moderate esophagitis was confirmed in 83 patients, Barrett's esophagus in 12 patients, and type I or II hiatal hernia in 13 patients. All patients were also submitted to manometry, 24 hour intraesophageal pH monitoring, and barium swallow before and after surgery. Follow-up ranged from 12 to 36 months. There were no conversion, major intraoperative, or postoperative complications; nor were there any deaths. Postoperative dysphagia was present in 5% of cases. Symptomatic recurrence of reflux was observed in 10.3% and endoscopic presence of esophagitis in 12.3% of cases . Lower esophageal sphincter pressure increased significantly after surgery, even in patients with endoscopic recurrence. 24-hour intraesophageal monitoring improved after surgery, except in patients with objective recurrence of esophagitis. In conclusion, laparoscopic cardial calibration with posterior gastropexy presents comparable results to those reported after Nissen fundoplication and therefore could be another excellent therapeutic option in patients with reflux esophagitis. PMID- 15827859 TI - Does a second-look operation improve survival in patients with peritonitis due to acute mesenteric ischemia? A five-year retrospective experience. AB - Second-look laparotomy is one of the mainstays of surgical treatment of acute mesenteric ischemia (AMI). The aim of this study was to analyze its role in the survival of patients with infarcted gangrenous bowel resulting from AMI. A retrospective chart review of all patients admitted over the study period was undertaken. The study population consisted of 41 patients with clinical evidence of peritonitis and gangrenous, perforated bowel on surgical exploration. Outcome was compared among patients who underwent second-look laparotomy and those who did not. Fifteen patients with an American Society of Anesthesiologists (ASA) score of less than 4 underwent second-look laparotomy. Six patients had residual necrotic bowel that required additional resection. Only one (17%) of them survived. Of the nine remaining patients, who had no evidence of necrosis, only two survived (22%). Overall survival in this group was 20%. Twenty-six patients were managed without second-look laparotomy. Nine of them, with an ASA score of 4 5, died soon after the operation. The decision not to operate on the remaining 17 patients with an ASA score < 4 was made by an experienced surgeon. Eleven of those patients (65%) survived. Overall survival in the non-second-look group was 42%. Excluding the early deaths, the survival in the non re-explored group was significantly higher than in the second-look group (65% vs. 20%, p = 0.011). A selective approach to the surgical treatment of acute mesenteric ischemia based on the sound clinical judgment of an experienced surgeon may be as appropriate as its universal application. PMID- 15827864 TI - Highlights in surgery through outstanding ISS/SIC surgeons: Russian surgeons of the Soviet Union: Was contact with the ISS/SIC feasible? PMID- 15827865 TI - Vascular risk factors in the Swiss population. AB - BACKGROUND AND PURPOSE: Identification of the population at risk of stroke remains the best approach to assess the burden of cardiovascular morbidity and mortality. METHODS: The prevalence of hypertension (HT), hypercholesterolemia (HCh), diabetes mellitus (DM), overweight (OW), obesity (OB), tobacco use (SM), and their combinations was examined in 4,458 Swiss persons (1,741 men and 2,717 women, mean age 57.8 +/- 15 years), who volunteered for the present survey. RESULTS: OW was the most prevalent risk factor (50 %), followed by HT (47%), HCh (33%), SM (13 %) and DM (1.6 %). The proportion of persons without risk factors (RF) was 19.9%, with 1 RF 41.5%, 2 RF 33.8%, 3 RF 4%, and 4 RF 0.9%. OW was more prevalent in men than in women (53% vs. 41%, P=0.02). More men than women aged 41 50 years and 51-60 years had HT (49 % vs. 36%, P=0.01, and 52 % vs. 42%, P=0.02). The prevalence of HCh and DM did not show any sex-related differences. HT, OW and HCh were not only the most common single risk factors, but were also most likely to aggregate with each other. CONCLUSIONS: The majority of Swiss people have one or two vascular risk factors. OW and HT are by far most common and are likely to aggregate with each other. A small modification of these two factors would reduce the incidence of stroke and myocardial infarction significantly. PMID- 15827866 TI - Peripheral neuropathy in CADASIL. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is a hereditary cerebral microangiopathy associated with mutations in the Notch 3 gene. The clinical phenotype is characterized by cerebral impairment even though typical microvascular changes are diffuse. OBJECTIVE: To assess peripheral neuropathy in patients with CADASIL. PATIENTS AND METHODS: We enrolled eleven CADASIL patients with variable phenotype including clinical signs of peripheral nerve involvement. In all patients electromyography and nerve conduction velocities were performed. Peripheral nerve biopsy was performed in three cases. RESULTS: We found sensory motor neuropathy in 7/11 patients. Nerve biopsy revealed axonal and demyelinated findings. CONCLUSION: Our findings suggest that peripheral neuropathy may be part of the CADASIL phenotype. PMID- 15827867 TI - Sensorineural hearing loss in patients with chronic progressive external ophthalmoplegia or Kearns-Sayre syndrome. AB - In the present study we assessed the prevalence and nature of hearing loss in patients with chronic progressive external ophthalmoplegia (CPEO) or Kearns-Sayre syndrome (KSS) due to single large-scale mitochondrial DNA (mtDNA) deletion or mtDNA tRNA (Leu (UUR)) A3243G point mutation (A3243G PM). 14 patients with mtDNA deletion and three patients with A3243G PM underwent audiological evaluation comprising pure-tone and speech audiometry as well as transient evoked otoacoustic emissions (OAE). Audiological evaluation revealed hearing impairment in 10/17 patients. Hearing loss was mild to moderate predominantly affecting high frequencies in five patients with subjective hearing problems (three patients with mtDNA deletions, two patients with A3243G PM). Subclinical hearing deficits restricted to high frequencies were seen in further five asymptomatic patients (four patients with mtDNA deletions, one patients with A3243G PM). Audiological findings suggested a cochlear origin of hearing loss in all subjects. Our results demonstrate that CPEO or KSS patients due to mtDNA deletion or A3243G PM are at high risk of developing sensorineural hearing deficits. PMID- 15827869 TI - Expanding the spectrum of Helicobacter pylori-associated diseases. PMID- 15827868 TI - The relation of the XbaI and PvuII polymorphisms of the estrogen receptor gene and the CAG repeat polymorphism of the androgen receptor gene to peak bone mass and bone turnover rate among young healthy men. AB - The genes coding for estrogen receptor-alpha (ER-alpha) and androgen receptors (AR) are potential candidates for the regulation of bone mass and turnover, which may contribute to both the achievement of peak bone mass and bone loss after completion of growth. The present study was aimed at elucidating the role of two restriction fragment lengths (XbaI and PvuII) polymorphisms of the ER gene and the CAG repeat polymorphism of the AR gene as determinants of peak bone mass in men; special attention was paid to the interaction between serum free estradiol (E2) levels and the XbaI and PvuII genotypes. A cross-sectional study, with data on lifestyle factors collected retrospectively, was performed in 234 young men, aged 18.3 to 20.6 years. Of the men, 184 were recruits of the Finnish Army and 50 were men of similar age who had postponed their military service for reasons not related to health. Bone mineral content (BMC), density (BMD) and scan area were measured in the lumbar spine and upper femur by dual-energy X-ray absorptiometry (DXA). The bone turnover rate was assessed by measuring serum type I procollagen aminoterminal propeptide (PINP) and tartrate-resistant acid phosphatase 5b (TRACP5b) as well as urinary excretion of type I collagen aminoterminal telopeptide (NTX). After adjusting for age, height, weight, exercise, smoking, calcium and alcohol intake, BMC, scan area and BMD at all measurement sites were similar for the different XbaI and PvuII genotypes of the ER and independent of the number of the CAG repeats of the AR gene. No association was found between free E2 levels and bone parameters among any genotype group of the XbaI and PvuII polymorphisms. Except for urinary NTX, which showed a tendency to higher values for the xx (P=0.08) and pp (P=0.10) genotypes of the ER, bone turnover markers were not related to the genotypes studied. Our study does not support the view that the XbaI and PvuII polymorphisms of the ER gene and the CAG polymorphism of the AR gene would have a substantial impact on the development of peak bone mass in young Finnish men. PMID- 15827870 TI - Mortality risk factors with nosocomial Staphylococcus aureus infections in intensive care units: results from the German Nosocomial Infection Surveillance System (KISS). AB - INTRODUCTION: As the number of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infections in German intensive care units increases, the problem of MRSA infection as such is becoming ever more serious. The aim of this study was to investigate whether mortality rates from nosocomial MRSA pneumonia and primary bloodstream infections (BSI) differ significantly from those of nosocomial pneumonia and primary BSI caused by methicillin-susceptible S. aureus (MSSA). METHODS: For the analysis data from the ICU component of the German nosocomial infection surveillance system (KISS) were used (January 1997 to June 2002). To identify mortality risk factors a logistic regression analysis with step-wise variable selection was conducted including all cases of nosocomial S. aureus pneumonia and primary BSI. The possible risk factors that were evaluated were age > median, male gender, time in the ICU before infection > median, type of ICU, type and size of hospital, intubation, CVC use, total parenteral nutrition, year of investigation, infection caused by MRSA. RESULTS: Data from 274 ICUs and 505,487 ICU patients were recorded and a total of 6,888 cases of nosocomial pneumonia and 2,357 cases of primary BSI identified, of which 1,851 cases of S. aureus pneumonia and 378 cases of S. aureus primary BSI were considered for analysis. 59 of the 349 patients with MRSA pneumonia (16.9%) and 105 of the 1,502 patients with MSSA pneumonia (7.0%) died. 16 of the 95 patients with primary MRSA BSI (16.8%) and 17 of the 283 patients with primary MSSA BSI died (6.0%). Four factors were significantly associated with mortality from S. aureus pneumonia, one of them being pneumonia caused by MRSA (OR = 2.62; CI95 1.69-4.02). Only MRSA was significantly associated with death from S. aureus primary BSI (OR = 3.84; CI95 1.51-10.2). CONCLUSION: Nosocomial pneumonia and primary BSI from MRSA may be associated with death, but the cause-effect relationship of severity of illness and MRSA remains to be determined due to the limitations of surveillance data. PMID- 15827871 TI - Coagulase-negative staphylococcal meningitis in adults: clinical characteristics and therapeutic outcomes. AB - BACKGROUND: We wanted to analyze the clinical characteristics and therapeutic outcomes of adult meningitis caused by coagulase-negative staphylococci (CoNS). PATIENTS AND METHODS: Over a period of 5 years (January 1999 to December 2003), 127 cases were identified as having adult culture-proven bacterial meningitis caused by a single pathogen. Of them, 14 cases with CoNS meningitis were enrolled, and their clinical characteristics and therapeutic outcomes were analyzed. RESULTS: The 14 cases (median age 37.5; range 24-77 years old) included nine men and five women. With polynerase chain reaction sequencing of bacterial 16S r-RNA, 10 of the 14 CoNS strains were identified as Staphylococcus epidermidis infection, and the other four belonged to Staphylococcus haemolyticus. All 14 cases were in a postneurosurgical state with insertion of a ventriculoperitoneal shunt, external ventricular device or intrathecal port A as their underlying conditions, and 12 of the 14 patients contracted the infection nosocomially. Fever (86%), leukocytosis (79%), hydrocephalus (50%), consciousness disturbance (36%), and seizure (7%) were the major clinical manifestations. All the involved CoNS strains showed resistance to oxacillin but retained their susceptibility to vancomycin and linezolid. All 14 CoNS strains had positive mecA gene detection. With the removal of neurosurgical devices and intravenous vancomycin therapy, 86% (12/14) of the patients survived. CONCLUSION: CoNS meningitis accounted for 11% (14/127) of our adult bacterial meningitis. All adult CoNS meningitis patients had a disrupted barrier of the central nervous system as the underlying condition. S. epidermidis was the most common CoNS subtype involved. All involved CoNS strains were oxacillin resistant. The therapeutic result showed that adult CoNS meningitis had a mortality rate of 14% (2/14). PMID- 15827872 TI - HIV infection among low-risk first lifetime testers in Rome, 1990-2000. AB - BACKGROUND: HIV spread among low-risk populations through heterosexual intercourse is a major public health concern. This study was aimed at describing prevalence and determinants of HIV infection among Italian low-risk subjects seeking their first lifetime HIV test. PATIENTS AND METHODS: Information collected between January 1990 and December 2000 at a major counseling and testing site in Rome, Italy, was analyzed. Multiple logistic regression odds ratios (OR) and 95% confidence intervals (CI) were computed. RESULTS: Among the 14,313 study subjects, 64 (0.4%) were seropositive for HIV infection. HIV seropositivity increased with age (OR = 4.0, 95% CI: 2.1-7.6 for >/= 40 years vs 18-24), and it seemed to be more common among men (OR = 1.6, lower 95% CI:0.9). There was no evidence of temporal variations, whereas motivations for HIV testing were strongly associated with HIV prevalence. Testing for alarming symptoms (OR = 13.8) or for heterosexual intercourse (OR = 11.0) were associated with a more than 10-fold increased HIV risk. CONCLUSION: Our findings are consistent with data from other industrialized countries and they show a strong association between HIV seropositivity and reason for first-time testing. Moreover, they indicate a stable trend of HIV prevalence among low-risk persons in the last decade. Further studies on time trends in low-risk populations would be useful to evaluate current HIV prevention programs. PMID- 15827873 TI - Detection of human hantavirus infections in Lithuania. AB - BACKGROUND: In Europe certain hantaviruses are known to cause hemorrhagic fever with renal syndrome of different severity. The objective of the present investigation was to study the presence of hantavirus infections in Lithuania. MATERIAL AND METHODS: Two different serum panels from cancer patients (n = 438) and blood donors (n = 299) from Lithuania were tested by monoclonal antibody capture IgG ELISA using yeast-expressed recombinant nucleocapsid (rN) proteins of Puumala virus (PUUV), Hantaan virus (HTNV) and Dobrava virus (DOBV). The reactivity of ELISA-positive sera was proven in Western blot tests using various hantavirus rN proteins. Selected serum samples were further analyzed by focus reduction neutralization assays. RESULTS: In the IgG ELISA 39 sera from the cancer patients and four sera from blood donors were found to be reactive with at least one of the rN proteins. By immunoblot using the three yeast-expressed rN proteins, the ELISA reactivity of 36 of 39 and two of four serum samples from cancer patients and blood donors, respectively, was confirmed; this corresponds to a seroprevalence of 8.2% and 0.7%, respectively. In ELISA, the majority of the samples reacted exclusively with rN proteins of HTNV and DOBV (31 of 36 and one of two in the two groups). In the group of sera selected for serotyping by focus reduction neutralization assay, this dominance was confirmed by the identification of eight DOBV but only four PUUV infections. No infection by HTNV or another hantavirus besides DOBV and PUUV was verified. Anti-hantavirus positive human sera were detected in all seven investigated counties of Lithuania. CONCLUSION: In Lithuania at least two hantaviruses, DOBV and PUUV, circulate and cause human infections. Additional investigations are needed to study the seroprevalence more precisely and to search for clinical cases of hantavirus infections. PMID- 15827874 TI - Typing of human cytomegalovirus clinical isolates from Saudi patients by PCR RFLP. AB - BACKGROUND: Information on strain types of human cytomegalovirus (HCMV) isolates from Saudi Arabian patients is lacking. MATERIALS AND METHODS: 80 clinical isolates of HCMV from Saudi Arabian patients were analyzed by PCR amplification of three regions (DNA polymerase, glycoprotein B, and glycoprotein H) of the virus genome. The resultant amplicons (2.0-2.7 kb) were further studied by restriction fragment length polymorphism (RFLP) using four enzymes (HaeIII, HhaI, MspI, and RsaI). RESULTS: Combined analysis of the cleavage patterns generated by the enzymes identified five strains, S1-S5, and several mixed and unique strains. 18 isolates belonged to S1 strain and were similar to laboratory strain AD169. Eight isolates were present in each of S2 and S3 strains. Six isolates and four isolates were found in S4 and S5 strains, respectively. 12 isolates contained a mixture of S3 and S5, which may have resulted from a dual infection. Each of the 24 remaining isolates had a different strain pattern. CONCLUSION: Our findings show that 80 HCMV clinical isolates were distributed into 30 different strains using PCRRFLP analysis of multiple viral subgenomic regions. However, the number of isolates is not uniformly distributed among strains (p < 0.02). PMID- 15827875 TI - Cultural recovery and determination of antimicrobial susceptibility in Helicobacter pylori by using commercial transport and isolation media. AB - BACKGROUND: Antimicrobial resistance of Helicobacter pylori is the main reason for eradication failure. We have studied the feasibility of a commercial transport medium for cultural recovery and subsequent drug susceptibility testing. PATIENTS AND METHODS: From March to December 2000, 79 consecutive gastric biopsies, positive in a rapid urease test, were transferred into a commercial transport medium and sent within 24 hours from the district hospital to the microbiological laboratory for culture and susceptibility testing. A commercial agar plate and an in-house Wilkins-Chalgren agar plate were used for culture. Susceptibility data were compared with data collected from 1992 to 2003 in the University Hospital of Zurich. RESULTS: Cultural recovery and susceptibility testing of H. pylori was successful in 55 of 79 patients. In 17 cases cultural recovery failed because of technical problems (n = 14), long transport time (n = 1) and unknown reason (n = 2). Failure of susceptibility testing (n = 7) was mainly due to fungal overgrowth. Resistance to metronidazole and clarithromycin was found in 15 (27%) and in 12 patients (22%), respectively; resistance to amoxicillin was not observed. Five patients (9%) showed resistance both to metronidazole and to clarithromycin. Eradication therapy failed in all patients with macrolide resistance. Resistance rates were higher in females than in males; 30% vs 12% for clarithromycin and 33% vs 20% for metronidazole. Resistance to metronidazole was significantly lower in Swiss patients (15%) than in non-Swiss patients (39%). CONCLUSION: Antimicrobial resistance data can reliably be obtained by sending the biopsy specimen in a commercial transport medium to a microbiological laboratory. This is especially important after eradication failure. Resistance to metronidazole and clarithromycin is highly prevalent and more common in women and non-Swiss patients. PMID- 15827876 TI - Helicobacter pylori infection in anterior uveitis. AB - BACKGROUND: Despite intensive research, the etiology of acute anterior uveitis (AAU) remains poorly defined. Infection with gram-negative bacteria such as Yersinia, Salmonella, Shigella, and Chlamydia have already been suggested as a possible trigger event for AAU. Helicobacter pylori is also a gram-negative bacterium, shares the lipopolysaccharides, but did not attract the attention of many ophthalmologists until recently. Having in mind the relatively high incidence of H. pylori infection in the population, we propose that H. pylori may also be a trigger factor for AAU. PATIENTS AND METHODS: The presence of anti-H. pylori antibodies in matching serum and aqueous humor samples of 15 idiopathic AAU patients was determined using a commercial Western blot assay. Control serum and aqueous humor were obtained from five patients undergoing cataract surgery. RESULTS: Six out of 15 AAU patients (40%) were serum-positive for H. pylori, and half of these (n = 3) also had anti-H. pylori antibodies in the aqueous humor. All five aqueous humor and sera controls tested negative for H. pylori infection. CONCLUSION: These are the first results demonstrating anti-H. pylori antibodies in the aqueous humor of AAU patients. Further studies are needed to demonstrate whether this antibody is indeed locally produced. Our data may provide first evidence for a causative link between H. pylori infection and AAU. PMID- 15827877 TI - Primary cutaneous cryptococcosis and secondary antigenemia in a patient with long term corticosteroid therapy. AB - We report a case of a 71-year-old male who developed severe cellulitis of his right forearm and hand after he had an accidental injury from the sharp edge of a metal plate of a birdhouse. The patient suffers from chronic asthma and has been treated with systemic corticosteroids for years. Culture of aspirates from two sites of the wound area revealed growth of Cryptococcus neoformans in one and Acinetobacter lwoffii in the other. After combined treatment including antibiotics, antifungal therapy with fluconazole 400 mg/d and surgical debridement followed by a mesh graft, the patient achieved complete healing of the wound. Five months after the infection, the patient was still positive for cryptococcal antigen at a titer of 1:64 despite oral treatment with fluconazole 50 mg/d, and maintenance therapy with fluconazole 200 mg/d was recommended for 6 months, or longer depending on further results. The clinical and microbiological characteristics of this patient as well as therapeutical and epidemiological aspects of primary cutaneous cryptococcosis (PCC) are discussed. PMID- 15827878 TI - Systemic aspergillosis with predominant genitourinary manifestations in an immunocompetent man: what we can learn from a disastrous follow-up. AB - Prostatic aspergillosis is a rare finding with only ten cases reported previously. We report the first case of systemic aspergillosis predominantly presenting with prostatic involvement and, clinically, with urinary retention in an immunocompetent host. Routine transurethral resection was performed due to benign prostatic hyperplasia with subvesical obstruction. Concomitant prostatic aspergillosis was diagnosed without signs of systemic infection. In the clinical follow-up systemic aspergillosis became rapidly progressive requiring complex surgical interventions and long-term antifungal therapy. The current literature is reviewed, and diagnostic and management options are discussed. PMID- 15827879 TI - Osteomyelitis and secondary septic arthritis caused by Sphingomonas paucimobilis. AB - We describe a case of osteomyelitis and secondary septic arthritis caused by Sphingomonas paucimobilis in an immunosupressed adolescent which did not present typically, leading to a delay in diagnosis. This case highlights the need to thoroughly investigate musculoskeletal pain in the immunocompromised patient in order to exclude an infection as a cause. PMID- 15827880 TI - Complications, thrombophlebitis rates, and intravenous catheter replacement strategies. PMID- 15827882 TI - Temporal trend in the level of 90K glycoprotein after HIV seroconversion among persons coinfected with hepatitis C virus. PMID- 15827884 TI - A quantitative analysis of optic nerve axons in elderly control subjects and patients with Alzheimer's disease. AB - OBJECTIVE: To study the density and cross-sectional area of axons in the optic nerve in elderly control subjects and in cases of Alzheimer's disease (AD) using an image analysis system. METHODS: Sections of optic nerves from control and AD patients were stained with toluidine blue to reveal axon profiles. RESULTS: The density of axons was reduced in both the center and peripheral portions of the optic nerve in AD compared with control patients. Analysis of axons with different cross-sectional areas suggested a specific loss of the smaller sized axons in AD, i.e., those with areas less that 1.99 microm2. An analysis of axons >11 microm2 in cross-sectional area suggested no specific loss of the larger axons in this group of patients. CONCLUSIONS: The data suggest that image analysis provides an accurate and reproducible method of quantifying axons in the optic nerve. In addition, the data suggest that axons are lost throughout the optic nerve with a specific loss of the smaller-sized axons. Loss of the smaller axons may explain the deficits in color vision observed in a significant proportion of patients with AD. PMID- 15827885 TI - The mode of spinal motor neurons degeneration in a model of slow glutamate excitotoxicity in vitro. AB - The defective glial and/or neuronal glutamate transport may, in chronic neurotoxicity, contribute to several neurodegenerative diseases including amyotrophic lateral sclerosis (ALS)--a progressive neurodegenerative disorder of lower and upper motor neurons (MNs). To determine the detailed ultrastructural characteristics of excitotoxic motor neurons neurodegeneration we used a model of slow excitotoxicity in vitro based on selective inhibition of glutamate uptake. The study was performed on organotypic cultures of the rat lumbar spinal cord subjected to various concentrations of glutamate uptake blockers: threohydroxyaspartate (THA) and L-trans-pyrrolidine-2, 4-dicarboxylate (PDC). The chronic inhibition of glutamate transport resulted in a dose-dependent slow neurodegeneration of spinal MNs consisting of necrotic, apoptotic and autophagic mode of cell death. There were some MNs that shared certain characteristics of a different type of cell injury. The results showed that a different mode of cell death in excitotoxic MNs degeneration may coexist resulting in apoptosis-necrosis and apoptosis-autophagocytosis continuum. PMID- 15827886 TI - The antiphospholipid syndrome in pregnant rabbits and their offspring. Neuropathological aspects. AB - OBJECTIVE: Although the obstetric consequences of antiphospholipid syndrome (APS) in pregnant rabbits have been described, there are no data on the serological and neuropathological aspects of the syndrome in their offspring. It would also be interesting to recognize whether the CNS abnormalities in rabbit fetuses relate to placental damage or depend on the antiphospholipid antibodies, transmitted from the pregnant animal through the placenta to the fetal serum. MATERIAL AND METHODS: A post-mortem neuropathological examination was done on 36 adult female New Zealand rabbits, and their offspring (100 fetuses). The material was divided into 4 groups: Group I--26 pregnant rabbits with experimental APS, Group IC 10- pregnant rabbits without APS (control group I), Group II--64 fetuses derived from animals included in Group I, and Group IIC--36 fetuses derived from individuals included in Group IC (control group II). The platelet count, activated partial thromboplastin time (APTT), antiplatelet antibodies in serum and coated on the platelets were evaluated to identify the APS in adult rabbits and their offspring. RESULTS: A significantly higher number of fetuses demonstrating weaker vitality and shorter survival time was observed in Group II. The percentage of dead and reabsorbed fetuses was also considerably higher in Group II. The serum markers of APS occurred both in Group I and II while the neuropathological evidences of APS: the thrombo-necrotic and inflammatory changes were found exclusively in APS pregnant animals. Moreover, cytoarchitecture of the fetal brains was intact. There were no disturbances in neuronal migration and abnormalities of cytodifferentiation. CONCLUSIONS: 1. The antiphospholipid syndrome in pregnant rabbits results in serological markers of the syndrome in their offspring. 2. The central nervous system of fetuses delivered from pregnant rabbits with the antiphospholipid syndrome remains intact despite the serological markers of the syndrome in fetus circulation. 3. The miscarriages in pregnant rabbits with the antiphospholipid syndrome depend rather on placental pathology related to the syndrome than on the syndrome per se transmitted from adult females to fetal circulation. PMID- 15827887 TI - Expression of apoptosis-related proteins in model of anoxia in vitro. AB - There has been growing evidence that different modes of cell death exist, among them the apoptosis is thought to be an important mechanism of nerve cell loss implicated in various pathological states. A number of proteins mediated with apoptotic process have been identified, including p53, BAX, BCL-2 and BCL-X. We examined the expression of proteins related to programmed cell death in hippocampal neurons in vitro, exposed to pure anoxia or pretreated with apoptosis modulating agents: zinc and zinc chelator - TPEN. The results evidenced the noticeable differences in the expression of pro- and anti-apoptotic proteins in particular experiments. In the cultures exposed to pure anoxia, a significant increase of p53 and BAX immunoreactivity, associated with the decreased level of BCL-2 and BCL-X immunopositive cells was observed, related to the activation of apoptotic process. Hippocampal cultures pretreated with ZnCl2 before anoxia showed decreased immunoreactivity for p53 and BAX, connected with BCL-2 overexpression, whereas the cultures exposed to zinc chelating agent - TPEN or TPEN connected with anoxia showed significant increase of immunorectivity for p53 and BAX. This strong immunoreactivity of proapototic proteins (p53 and BAX) in hippocampal cultures exposed to anoxia or/and TPEN correlated with previous ultrastructural evidences of anoxia- and TPEN-induced apoptosis, while the overexpression of anti-apoptotic protein (BCL-2 and BCL-X) in zinc-pretreated cultures evidenced the protective ability of this metal against apoptosis in model of anoxia in vitro. PMID- 15827888 TI - The neuroprotective function of vascular endothelial growth factor (VEGF). AB - The vascular endothelial growth factor (VEGF) is known mainly as the potent angiogenic and vascular permeability-enhancing factor. Both processes are very effective in hypoxia. The latest studies show that VEGF has neurotrophic and neuroprotective as well as angiogenic properties. It exerts neuroprotective actions directly through the inhibition of programmed cell death (PCD), or apoptosis and the stimulation of neurogenesis. VEGF is also a mediator of multiple processes including angiogenesis, enhancing blood brain barrier permeability for glucose, antioxidants activation, which indirectly result in neuroprotection. VEGF prevents neurons from death under critical conditions such as hypoxia, glucose deprivation through binding to the specific receptors, which are also expressed on the surface of neuronal cells. The increased expression of VEGFR-2/flk-1/KDR receptors on neurons subjected to hypoxia, glucose deprivation provides evidence that these receptors are mainly involved in neuroprotective effects of VEGF. Furthermore, binding to these receptors triggers the phosphatidyloinositol 3-kinase (PI3K) /Akt signal transduction system and, in consequence, leads to the inhibition of PCD by activating antiapoptotic proteins through the transcription factor NFkappaB and inhibiting proapoptotic signaling by Bad, caspase-9, caspase-3, and other effectors. Promotion of neuronal cells proliferation by VEGF is also associated with the increased expression of VEGFR-2 receptors and up-regulation of E2F family transcription factors, cyclin D1, cyclin E, and cdc25. It is known that the amount and types of VEGF isoforms influence its action. At least six isoforms of VEGF proteins are formed as a result of alternative mRNA splicing and it is unknown which of them and in what proportion occur in the nervous system in physiology and pathophysiology. It seems to be very essential to find out the mechanisms responsible for specific patterns of VEGF isoforms and their receptors expression in different pathologies of the nervous system. Maybe such knowledge will provide new perspectives in VEGF therapy. PMID- 15827889 TI - Brain abscess as the first clinical manifestation of multiple pulmonary arteriovenous malformations in a patient with hereditary hemorrhagic telangiectasia (Rendu-Osler-Weber disease). AB - In this report we described a case of a cerebral abscess that developed in presence of asymptomatic pulmonary arteriovenous malformations (PAVMs) in a 53 year-old woman with hereditary hemorrhagic telangiectasia (HHT). The brain abscess was aspirated with good clinical result and the arteriovenous fistulae qualified for transcatheter embolotherapy. Each patient suspected to suffer from HHT should be diagnosed for the presence of visceral vascular malformations. Most of them are found in the lungs, liver and brain. Early diagnosis and treatment of PAVM prevent the occurrence of severe neurological complications such as brain stroke or brain abscess. Cases of a cerebral abscess in adults of unexplained etiology should raise the suspicion of an asymptomatic PAVM. PMID- 15827890 TI - Recurrent cerebral infarcts as the first manifestation of infection with the HIV virus. AB - There is an increased risk of stroke in patients with HIV infection. One of the mechanisms is production of anticardiolipin (aCL) antibodies, induced by the virus. Many studies have documented a high incidence of aCL antibodies in patients with HIV infection and the increased risk of stroke, although there is no such correlation with other viruses that also stimulate production of aCL antibodies. Probably the HIV virus also stimulates production of other serum proteins, which together with aCL antibodies make procoagulation complexes, similar to the primary anticardiolipin syndrome. We present a case of a 40-year old patient, hospitalized three times in our department because of recurrent ischemic strokes. Diagnostic tests revealed a high titre of anticardiolipin antibodies and the early stage of HIV infection. Recurrent ischemic strokes were the first manifestation of HIV infection in this patient. PMID- 15827892 TI - Dyslipidemia and nephrotic syndrome: recent advances. AB - Patients with nephrotic syndrome (NS) have one of the most pronounced secondary changes in lipoprotein metabolism known, and the magnitude of the changes correlates with the severity of the disease. These changes are of a quantitative as well as a qualitative nature. All apolipoprotein B (apo B)-containing lipoproteins, such as very-low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), low-density lipoproteins (LDL), and lipoprotein(a) [Lp(a)], are elevated in nephrotic syndrome. High-density lipoproteins (HDL) are reported to be unchanged or reduced. In addition to these quantitative changes, the lipoprotein composition is markedly changed, with a higher ratio of cholesterol to triglycerides in the apo B-containing lipoproteins and an increase in the proportion of cholesterol, cholesterol ester, and phospholipids compared with proteins. Also apolipoproteins show major changes, with an increase in apolipoprotein A-I, A-IV, B, C, and E. Particularly the changes in apo C-II, which is an activator of the enzyme lipoprotein lipase (LPL), and apo C-III, an inhibitor of LPL, with an increase of the C-III to C-II ratio, might contribute to the impaired lipoprotein catabolism in NS. The mechanisms for these changes in lipoprotein metabolism are discussed in this review as far as they are known. Furthermore, the tremendous elevations of Lp(a) in nephrotic syndrome and its primary and secondary causes are reviewed. Primary causes became recently apparent by a significantly higher frequency of low-molecular-weight apo(a) phenotypes in patients compared with controls. The secondary causes were shown by an increase of Lp(a) in all apo(a) isoform groups. Because Lp(a) is an LDL-like particle that is usually included in the measured or calculated LDL cholesterol fraction, the influence of the extremely high Lp(a) levels in NS on the measurement of LDL cholesterol is discussed. PMID- 15827893 TI - Prevalence of dietary supplement use in adults with chronic renal insufficiency. AB - OBJECTIVE: There are currently no studies that have investigated dietary supplement use in the chronic renal insufficiency (CRI) population. The objectives of this study were to determine the prevalence of dietary supplement use in a sample of adults with CRI and to examine whether the prevalence of dietary supplement use was higher in the earlier stages of CRI. DESIGN AND SETTING: Cross-sectional survey conducted at an outpatient nephrology clinic. PATIENTS: Adults (n = 100) with varying degrees of CRI. INTERVENTION: Interview administered questionnaire. MAIN OUTCOME MEASURES: The questionnaire contained questions related to the use of dietary supplements, attitudes, and beliefs about dietary supplements, as well as medical and demographic data including gender, age, body weight, smoking status, concurrent chronic conditions, prescribed medications, serum creatinine, estimated glomerular filtration rate (GFR), etiology of CRI, income, and education level. RESULTS: The prevalence of dietary supplement use was 45%, with the most frequent use occurring in subjects with a mild to moderate degree of renal insufficiency. There were no significant differences in gender, age, body mass index, smoking status, number of concurrent chronic conditions, number of prescribed medications, income, or education level between supplement users and supplement nonusers. Further analysis of dietary supplement use showed that the number of supplements used ranged from 1 to 5 or more supplements per day, with an overall total of 36 different supplements. Dietary supplement users were significantly more concerned with their health than were supplement nonusers. The most frequent sources of supplement information were physicians and printed material; however, 75% of the dietary supplement users desired further information about their dietary supplements. CONCLUSION: This is the first study to investigate the use of dietary supplements in adults with CRI, and it provides valuable information for health care professionals regarding the dietary supplement use of their patients with CRI. PMID- 15827894 TI - Assessment of nutritional status in hemodialysis patients using patient-generated subjective global assessment. AB - OBJECTIVE: To evaluate the scored Patient-Generated Subjective Global Assessment (PG-SGA) as a nutrition assessment tool in hemodialysis patients. DESIGN: A cross sectional observational study assessing the nutritional status of hemodialysis patients. SETTING: Private tertiary Australian hospital. SUBJECTS: Sixty patients, ages 63.9 +/- 16.2 years. INTERVENTION: Scored PG-SGA questionnaire, comparison of PG-SGA score > or =9 with subjective global assessment (SGA), albumin, corrected arm muscle area, and triceps skinfold. RESULTS: According to SGA, 80% of patients were well nourished and 20% of patients were malnourished. Patients classified as well nourished (SGA-A) attained a significantly lower median PG-SGA score compared with those rated as moderately malnourished or at risk of malnutrition (SGA-B). A PG-SGA score > or =9 had a sensitivity of 83% and a specificity of 92% at predicting SGA classification. There were significant correlations between the PG-SGA score and serum albumin, PG-SGA score, and percentage weight loss over the past 6 months. There was no association between PG-SGA score and body mass index or anthropometric measurements. CONCLUSION: The scored PG-SGA is an easy-to-use nutrition assessment tool that allows quick identification of malnutrition in hemodialysis patients. PMID- 15827895 TI - Physical activity patterns in chronic hemodialysis patients: comparison of dialysis and nondialysis days. AB - OBJECTIVE: To determine physical activity patterns in chronic hemodialysis patients with a specific emphasis on the difference between dialysis and nondialysis days. Design A cross-sectional single-center study. SETTING: Vanderbilt University Outpatient Dialysis Unit. PATIENTS: Twenty current chronic hemodialysis patients: 10 male, 10 female; 15 black, 5 white; mean age, 50.1 +/- 9.9 years; height, 164.5 +/- 10.9 cm; weight, 82.5 +/- 15.4 kg; length on dialysis, 57.3 +/- 45.3 months. METHODS: Minute-by-minute physical activity was assessed over a 7-day period using a triaxial accelerometer, which consists of raw numbers or counts calculated by the 3 axes of the accelerometer (PA counts). PA counts were extrapolated on a daily and hourly basis. Physical functioning tests included: sit-to-stand, 6-minute walk, and 1-repetition maximal leg press exercise. Laboratory values for serum concentrations of albumin, prealbumin, C reactive protein, and cholesterol were also collected. MAIN OUTCOME MEASURE: PA counts. RESULTS: Total PA counts were significantly lower on dialysis days when compared with nondialysis days (128,279 +/- 74,009 versus 168,744 +/- 95,168, respectively, P = .025). The average PA counts during the 4-hour dialysis time period were significantly lower on dialysis days when compared with nondialysis days (3,086 +/- 3,749 versus 11,070 +/- 7,695, respectively, P = .001). At postdialysis hours 1 and 2, PA counts on dialysis days were significantly higher than on nondialysis days (11,410 +/- 5,340 versus 9,082 +/- 6,646, P = .008, and 14,048 +/- 9,728 versus 8,662 +/- 6,433, P = .016, respectively). By postdialysis hour 4, PA counts on dialysis days had significantly decreased when compared with nondialysis days (6,068 +/- 6,268 versus 10,512 +/- 7,420 PA counts, P = .01, respectively). From postdialysis hours 5 to 20, there was no significant difference in PA counts between dialysis and nondialysis days. CONCLUSION: This study shows that physical activity is lower on dialysis days when compared with nondialysis days, and this decrease is caused by the lack of activity during the 4-hour hemodialysis procedure. New behavior modification strategies involving physical activity, both during hemodialysis and on nondialysis days, must be examined in this patient population. PMID- 15827896 TI - L-carnitine infusions may suppress serum C-reactive protein and improve nutritional status in maintenance hemodialysis patients. AB - Scattered reports indicate that L-carnitine may suppress proinflammatory cytokines in sick individuals without renal disease and may improve protein synthesis or nitrogen balance either in patients without renal disease or in maintenance hemodialysis (MHD) or chronic peritoneal dialysis patients. We conducted an experimental study in MHD patients to evaluate the effects of L carnitine treatment on inflammatory and protein-energy nutritional status. MHD patients were assigned to receive intravenous injections of L-carnitine 20 mg/kg (n = 48) or placebo (n = 65) thrice weekly at the end of each hemodialysis treatment for 6 months. The carnitine-treated group showed a statistically significant decrease in serum C-reactive protein and increase in serum albumin and transferrin, blood hemoglobin, and body mass index. Conversely, in the placebo-treated group, a significant decrease was reported for serum albumin, serum transferrin, and body mass index, whereas the other considered measures did not change significantly. These preliminary findings suggest that in MHD patients, L-carnitine therapy may suppress inflammation, particularly among those patients with C-reactive protein > or =3 mg/dL, and may improve protein-energy nutritional status. PMID- 15827897 TI - The Nutritional and Inflammatory Evaluation in Dialysis patients (NIED) study: overview of the NIED study and the role of dietitians. AB - The absolute majority of maintenance hemodialysis (MHD) patients die within 5 years of commencing dialysis treatment, mostly because of cardiovascular (CV) disease. The strongest and most common correlates of death in MHD patients are not conventional CV risk factors, but markers of protein-energy malnutrition and inflammation, together also known as malnutrition-inflammation complex syndrome (MICS). Paradoxically, classic risk factors such as obesity and hypercholesterolemia are associated with better survival in MHD patients. It has been hypothesized that this so-called reverse epidemiology is caused by the overwhelming prevalence and dominating effect of MICS in MHD patients. Hence, the key to improving survival and quality of life in MHD patients may be a better understanding of MICS and its interactions with CV disease and outcome. The Nutritional and Inflammatory Evaluation in Dialysis Patients (NIED) study is a longitudinal multicenter cohort study that aims to examine these hypotheses. At any given semiannual round, approximately 360 MHD patients from 8 DaVita dialysis facilities in the Los Angeles area are examined; 900 MHD patients will be cumulatively studied by the end of this 5-year prospective study (October 2001 to September 2006). Repeated measures of markers of nutritional status and inflammation are performed by 10 to 12 dialysis unit dietitians while patients attend their routine HD treatment in their dialysis facilities. All-cause and CV mortality, hospitalization, and quality of life are studied as outcome measures. The collaborating dietitians are the main evaluators and play crucial roles in all aspects of the study. This article reviews the design and infrastructure of the NIED study and reports preliminary findings of the first 12 to 30 months of the study. PMID- 15827898 TI - Variables associated with reduced dietary intake in hemodialysis patients. AB - BACKGROUND: Among the causes of malnutrition in hemodialysis (HD) patients, inadequate dietary intake (IDI) seems to be one of the most frequent and important. Although it has been hypothesized that IDI might be secondary to uremia, anorexia, underlying illness, psychosocial conditions, loss of dentures, depression, aging, or chronic inflammation, definite data on the etiology of IDI in HD patients are still lacking. The goal of this study was to measure the actual dietary energy and protein intakes in stable HD patients and to evaluate which demographic, clinical, dialytic, and humoral variables were associated with a dietary intake lower than recommended by international guidelines. METHODS: Thirty-seven patients maintained on regular HD, 3 times per week for 4 hours per session, were included in the study. In addition to epidemiologic data, patients were scrutinized for dry weight, weight change in the last 6 months, height, Body Mass Index, Kt/V, serum leptin, leptin-BMI ratio, presence of anorexia, and dietary energy and protein intake. Anorexia was assessed by means of a questionnaire in which the presence of major symptoms, namely meat aversion, taste and smell alterations, nausea and/or vomiting, and early satiety, was investigated. Dietary intake was recorded for 3 days after questionnaire administration by means of 3-day diet diaries. RESULTS: Overall, the mean (+/- standard deviation) dietary energy and protein intakes were 24.9 +/- 10.1 kcal/kg/day and 0.64 +/- 0.4 g protein/kg/day, respectively. Twenty-six patients (70.2%) had energy and protein intakes lower than recommended, 7 (18.9%) had adequate energy intake but inadequate protein intake, 1 (2.7%) had adequate protein intake and inadequate energy intake, and 3 (8.1%) had both adequate energy and adequate protein intakes. Anorexia was present in 14 of the 26 (53%) patients with low protein and energy intakes, and was absent in the other groups ( P =.003). The age of patients with inadequate energy and protein intakes was significantly higher than the age of patients with adequate energy and protein intakes (62.1 +/- 10.4 versus 37 +/- 20.8, P <.001) and the age of patients with only adequate energy intake (40.5 +/- 10.4, P <.001). Twenty-seven patients (73%) had an energy intake <30 kcal/kg/day, and 10 (27%) had an energy intake > or =30 kcal/kg/day. Compared with patients with energy intakes > or =30 kcal/kg/day, patients with energy intakes <30 kcal/kg/day were significantly older ( P =.0001) and more frequently were anorexic (P <.05). Compared with patients with protein intakes > or =1.2 g/kg/day, patients with protein intakes <1.2 g/kg/day were significantly older (P <.001). Limiting the analysis to the 33 patients with protein intakes <1.2 g/kg/day, we found a significant negative correlation between age and energy intake ( r =-0.612; P <.001) and between age and protein intake ( r =-0.723; P <.001). Correlations between both energy and protein intakes and age, dialytic age, Kt/V, C-reactive protein, parathyroid hormone, and leptin-BMI were not statistically significant. CONCLUSIONS: This study shows that dietary energy and protein intakes are inadequate in the majority of HD patients and are negatively related to the presence of anorexia and age. These data may be potentially useful in the identification of nutritional strategies as well as in improving food intake in HD patients. PMID- 15827899 TI - Short-term administration of omega-3 fatty acids in hemodialysis patients with balanced lipid metabolism. AB - BACKGROUND: At present, it is discussed whether omega-3 fatty acids show anti inflammatory, antithrombogenic, and antiatherosclerotic effects, also in patients with chronic renal failure. METHODS: In this prospective study, 11 hemodialysis (HD) patients, ages 59 +/- 17 years, who had balanced lipid metabolism and had been on HD for 53 +/- 47 months, were treated with a moderate dose of omega-3 fatty acids (1.2 g/day combined with 11.2 g/day pectin) for 12 weeks. Serum concentrations of c-reactive protein, homocysteine (Hcy), lipids, complement factors, blood gas analyses, 24-hour blood pressure, heart rate variability, electrocardiography, shunt blood flow, and recirculation, as well as peripheral oxygen saturation at the hand and foot, were measured at the start (t0w), and after 12 weeks (t12w) of therapy. Results Several assessed cardiovascular risk factors were significantly influenced. Levels of very-low-density lipoproteins (t0w, 77 +/- 26; t12w, 63 +/- 32 mg/dL; P <.05) and triglycerides (t0w, 261 +/- 157; t12w, 228 +/- 131 mg/dL; P =.068) were decreased. However, Hcy concentrations increased from 35.5 +/- 32.5 to 43.5 +/- 36.7 micromol/L ( P <.01) after 12 weeks. Anti-inflammatory and investigated clinical parameters did not significantly change during the study period. CONCLUSION: Limited positive effects on metabolic parameters were evaluated by short-term administration of omega-3 fatty acids in HD patients. Based on previous studies and on suspicion of atherosclerotic disorder in examined HD patients, we suppose that only high doses of omega-3 fatty acids given for a longer time influence inflammation and atherosclerosis. PMID- 15827900 TI - Bone disease and bottle caps. PMID- 15827901 TI - Protein supplementation in patients using peritoneal dialysis. PMID- 15827902 TI - K/DOQI practice guidelines for bone metabolism and disease in chronic kidney disease: another opportunity for renal dietitians to take a leadership role in improving outcomes for patients with chronic kidney disease. PMID- 15827905 TI - Go the distance. PMID- 15827906 TI - Accepting the gavel. PMID- 15827908 TI - Development of a clinical prediction rule for the diagnosis of carpal tunnel syndrome. AB - OBJECTIVES: To develop a clinical prediction rule (CPR) and to assess the reliability and diagnostic accuracy of individual clinical examination items for the diagnosis of carpal tunnel syndrome (CTS). DESIGN: Prospective diagnostic test study with blind comparison to a reference criterion of a compatible clinical presentation and abnormal electrophysiologic findings. SETTING: Multicenter medical center and community hospital with patient referrals from ambulatory primary care and specialty practice settings. PARTICIPANTS: Eight-two consecutively referred patients (50% men; mean age, 45+/-12 y) with suspected cervical radiculopathy or CTS referred for electrophysiologic examination. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Sensitivity, specificity, and likelihood ratios. RESULTS: The CPR identified in this study consisted of 1 question (shaking hands for symptom relief), wrist-ratio index greater than .67, Symptom Severity Scale score greater than 1.9, reduced median sensory field of digit 1, and age greater than 45 years. The likelihood ratio for the CPR was 18.3 when all 5 tests were positive. Interrater reliability was acceptable for all but 2 clinical examination items. CONCLUSIONS: The CPR identified was more useful for the diagnosis of CTS than any single test item and resulted in posttest probability changes of up to 56%. Further investigation is required both to validate the test-item cluster and to improve point-estimate precision. PMID- 15827909 TI - Effectiveness of a community-based Tai Chi program and implications for public health initiatives. AB - OBJECTIVES: To establish whether the reported beneficial physiologic effects of Tai Chi when performed under stringent experimental conditions can be generalized to the community. DESIGN: Phase 1: pre-post comparison in a group inexperienced in Tai Chi. Phase 2: baseline comparison between inexperienced and experienced Tai Chi groups. SETTING: A community in Hong Kong. PARTICIPANTS: Phase 1: 51 subjects inexperienced in Tai Chi (novice group) participated in the program. Phase 2: baseline measures of the novice group were compared with those of an experienced group (n=49) who had practiced Tai Chi for at least 6 months. INTERVENTION: A Cheng 119 style program was taught by a Tai Chi master for 1.5 hours, 3 times weekly, for 12 weeks. MAIN OUTCOME MEASURES: Lung function and physical activity evaluated before and after the completion of the program. Resting heart rate, blood pressure, oxygen saturation, handgrip strength, flexibility, and balance measured at the program commencement, 6 weeks, and 12 weeks. RESULTS: Phase 1: after the program, the novice group had increased handgrip strength, flexibility, and peak expiratory flow rate. Phase 2: the experienced group had greater flexibility, lower resting heart rate but higher diastolic blood pressure than the novice group prior to training. CONCLUSIONS: A community-based Tai Chi program produces beneficial effects comparable to those reported from experimental laboratory trials of Tai Chi; therefore, it should be considered as a public health strategy. PMID- 15827910 TI - Efficacy of a physical therapy program in patients with Parkinson's disease: a randomized controlled trial. AB - OBJECTIVE: To investigate the effects of a physical therapy (PT) program in groups of people with Parkinson's disease (PD). DESIGN: Randomized controlled trial with a crossover design. SETTING: Two outpatient rehabilitation clinics in Boston and Amsterdam, respectively. PARTICIPANTS: Sixty-eight subjects diagnosed with typical, idiopathic PD, Hoehn and Yahr stage II or III, and stable medication use. INTERVENTION: Group A received PT and medication therapy (MT) for the first 6 weeks, followed by MT only for the second 6 weeks. Group B received only MT for the first 6 weeks and PT and MT for the second 6 weeks. MAIN OUTCOME MEASURES: The Sickness Impact Profile (SIP-68), the mobility portion of the SIP 68, the Unified Parkinson's Disease Rating Scale (UPDRS), and comfortable walking speed (CWS) at baseline, 6-week, 12-week, and 3-month follow-up. RESULTS: At 6 weeks, differences between groups were significant for the SIP mobility ( P =.015; effect size [ES]=.55), for CWS ( P =.012; ES=.49), for the activities of daily living (ADL) section of the UPDRS ( P =.014; ES=.45), and for the total UPDRS ( P =.007; ES=.56). The total SIP and the mentation and motor sections of the UPDRS did not differ significantly between groups. Significant differences were found at 3 months compared with baseline for CWS, the UPDRS ADL, and total scores. CONCLUSIONS: People with PD derive benefits in the short term from PT group treatment, in addition to their MT, for quality of life related to mobility, CWS, and ADLs; long-term benefits were found in CWS, UPDRS ADL, and total scores but varied between groups. PMID- 15827911 TI - Bilateral changes in somatosensory sensibility after unilateral below-knee amputation. AB - OBJECTIVE: To evaluate possible alteration in proprioceptive and cutaneous sensibility in the nonamputated leg of unilateral transtibial amputees. DESIGN: Cross-sectional study with between-subjects (amputees vs controls) and within subjects (nonamputated vs amputated leg) comparisons. SETTING: Canadian rehabilitation hospital research laboratory. PARTICIPANTS: Two groups of amputees (34 due to traumatic causes, 14 due to vascular causes), recruited more than 1 year after their prosthetic training; and 2 groups (n=34, n=14) of age-matched control subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Threshold of movement detection and touch-pressure perception at the knee and foot levels. RESULTS: In the traumatic group, the sensory thresholds of the nonamputated leg were significantly higher than the control values in the 2 modalities tested. The movement detection was reduced at the knee and ankle levels, whereas a decrease in touch-pressure sensibility was observed only at the plantar site. As expected, a large proportion of the vascular amputees presented with severe sensory deficits in the nonamputated leg, particularly a loss in touch-pressure perception at the foot. The thresholds of movement detection were similar and correlated at both knees in the 2 groups of amputees. For the touch-pressure thresholds, no significant relationship was found between sides at the knee level. CONCLUSIONS: Sensory changes observed in the nonamputated leg suggest that central sensory adaptations occur after amputation. For movement detection, they were marked by a matching of perception on both sides of the body. Functional significance of these changes remains to be determined. PMID- 15827912 TI - Regional differences in pain threshold and tolerance of the transtibial residual limb: including the effects of age and interface material. AB - OBJECTIVES: To compare the pain threshold (the minimum pressure inducing pain) and pain tolerance (the maximum tolerable pressure) of different regions of the residual limbs of amputees by the indentation method and to evaluate the interface pressure distribution and distortion of the skin surface on indentation by finite element (FE) analysis. DESIGN: Crossover trial. SETTING: Rehabilitation engineering center. PARTICIPANTS: Eight transtibial amputees for indentation test and 1 for FE analysis. INTERVENTIONS: The load applied to the residual limbs using a Pelite or polypropylene indenter attached to a force transducer was increased until subjects could no longer tolerate the load. An FE model was built to simulate the indentation process with the experimentally recorded pain threshold used to load the indenters against the soft tissues. MAIN OUTCOME MEASURES: Pain threshold and tolerance and interface pressure and distortion of soft tissues. RESULTS: The patellar tendon and distal end of the fibula were the best and the worst load-tolerant regions, respectively. Some regions with a thicker layer of soft tissue had lower pain thresholds and tolerance than those with a thinner tissue layer. There was a trend for pain threshold and tolerance to decrease with age. The FE model showed that the peak pressure at the skin surface was very close when both indenters were loaded against the soft tissue at pain threshold limit. CONCLUSIONS: Contrary to common beliefs, regions with a thicker layer of soft tissue did not have a higher load-tolerant ability than thin-skinned regions. Pain threshold and tolerance could be age dependent. The FE model suggests that pain is triggered when peak pressure is applied to the residual limb exceeding a certain limit. PMID- 15827913 TI - Depressive symptoms and mental health service utilization among persons with limb loss: results of a national survey. AB - OBJECTIVE: To describe the prevalence of depressive symptoms, risk factors, and mental health service utilization in a national limb-loss sample. DESIGN: Cross sectional survey. SETTING: Participants were interviewed by telephone. PARTICIPANTS: A stratified sample by etiology of 914 persons with limb loss, derived from people who contacted the Amputee Coalition of America from 1998 to 2000. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Center for Epidemiologic Study Depression Scale (CES-D 10-item), pain bothersomeness, characteristics of the amputation, sociodemographics, and mental health service utilization. RESULTS: Prevalence for significant depressive symptoms (CES-D score, >/=10) was 28.7%. Risk factors included being divorced or separated, living at the near-poverty level, having comorbid conditions, being somewhat bothered or extremely bothered by back pain and phantom limb pain, and having residual limb pain for persons aged 18 to 54. Having higher education was a buffer against depressive symptoms. Almost 22% of the sample and 44.6% of persons with significant depressive symptoms received mental health service in the previous year. For persons with significant depressive symptoms, 32.9% reported needing mental health service but not receiving them, and 67.1% reported not needing mental health service. CONCLUSIONS: Depressive symptoms are prevalent among persons with limb loss. Proper management of pain and medical comorbidity may mitigate depressive symptoms. Education about depressive symptoms and treatment options may improve receipt of mental health service among persons with limb loss reporting significant levels of depressive symptoms. PMID- 15827914 TI - Dermatologic conditions associated with use of a lower-extremity prosthesis. AB - OBJECTIVES: To document the frequency of skin problems among lower-limb prosthesis users and to assess for factors associated with skin problems among patients using a prosthesis. DESIGN: Six-year retrospective chart review. SETTING: An outpatient amputee clinic at a regional, referral rehabilitation hospital in Canada. PARTICIPANTS: Seven hundred forty-five subjects with a total of 828 lower-extremity amputations participated. Subjects were included if they had a lower-extremity amputation and used a prosthesis for ambulation or transfers. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The presence or absence of a skin problem. Descriptive and nonparametric statistics were used to analyze data. RESULTS: Three hundred thirty-seven residual limbs (40.7%) had at least 1 skin problem. Adjusted odds ratios showed that amputation level, being employed, type of walking aid, and absence of peripheral vascular disease (as a comorbidity) were independently associated with the presence of at least 1 skin problem ( P <.05). CONCLUSIONS; Dermatologic conditions are a frequent complication for the lower-extremity amputee who uses a prosthesis. The results suggest that more active amputees have an increased risk for developing skin problems. Further study in this area is warranted. PMID- 15827915 TI - Dietary intake and nutritional status of urban community-dwelling men with paraplegia. AB - OBJECTIVES: To evaluate nutritional status, dietary intake, nutrition knowledge, and depression of healthy urban men with chronic spinal cord injury (SCI) and to compare these findings with national guidelines and data. DESIGN: Cross sectional. SETTING: Urban university. PARTICIPANTS: Ninety-five community dwelling men with paraplegia (age range, 20-59 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Dietary intake, body mass index (BMI), waist circumference, knowledge of nutrition, and depression. RESULTS: Diets included too much total and saturated fat, and inadequate fiber, calcium, fruit, and dairy intake. Most participants met protein needs, but most calorie levels were at or below recommendations. By using standard BMI and waist circumference cut-points for the able-bodied, approximately half of participants were overweight, 19% were obese, 7.5% were underweight, and more than one third had large waist circumferences. Participants with low knowledge of nutrition and high BMI who lived alone, smoked, and who had low family incomes were at significantly higher risk for lower quality diets. African Americans had the poorest diets. CONCLUSIONS: Intake of several key nutrients did not meet guidelines, and many BMI and waist circumference values were outside recommended ranges. These data highlight the need for clinicians to screen, counsel, and treat people with SCI to prevent related chronic diseases. PMID- 15827916 TI - Effectiveness of automated locomotor training in patients with chronic incomplete spinal cord injury: a multicenter trial. AB - OBJECTIVE: To determine whether automated locomotor training with a driven-gait orthosis (DGO) can increase functional mobility in people with chronic, motor incomplete spinal cord injury (SCI). DESIGN: Repeated assessment of the same patients or single-case experimental A-B design. SETTING: Research units of rehabilitation hospitals in Chicago; Heidelberg, Germany; and Basel and Zurich, Switzerland. PARTICIPANTS: Twenty patients with a chronic (>2 y postinjury), motor incomplete SCI, classified by the American Spinal Injury Association (ASIA) Impairment Scale with ASIA grades C (n=9) and D (n=11) injury. Most patients (n=16) were ambulatory before locomotor training. INTERVENTION: Locomotor training was provided using robotic-assisted, body-weight-supported treadmill training 3 to 5 times a week over 8 weeks. Single training sessions lasted up to 45 minutes of total walking time, with gait speed between .42 and .69 m/s and body-weight unloading as low as possible (mean +/- standard deviation, 37%+/ 17%). MAIN OUTCOME MEASURES: Primary outcome measures included the 10-meter walk test, the 6-minute walk test, the Timed Up & Go test, and the Walking Index for Spinal Cord Injury-II tests. Secondary measures included lower-extremity motor scores and spastic motor behaviors to assess their potential contribution to changes in locomotor function. All subjects were tested before, during, and after training. RESULTS: Locomotor training using the DGO resulted in significant improvements in the subjects' gait velocity, endurance, and performance of functional tasks. There were no significant changes in the requirement of walking aids, orthoses, or external physical assistance. There was no correlation between improvements in walking speed or changes in muscle strength or spastic motor behaviors. CONCLUSIONS: Intensive locomotor training on a treadmill with the assistance of a DGO results in improved overground walking. PMID- 15827917 TI - Return to work after spinal cord injury in Taiwan: the contribution of functional independence. AB - OBJECTIVE: To study the employment rate and determinants of return to work for persons with traumatic spinal cord injury (SCI) in Taiwan. DESIGN: Cross sectional. SETTING: Taiwan community. PARTICIPANTS: One hundred sixty-nine people who had sustained traumatic SCI, had been completely rehabilitated in a university hospital between 1989 and 2002, and who were between 18 and 60 years of age at the time of interview in 2003. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A structured questionnaire was used to identify demographic and injury-related status, functional status, and pre- and postinjury work-related information. RESULTS: At the time of survey, only 47% of the participants were engaged in remunerative employment. Cox regression analysis, with time elapsed between injury and survey as the underlying time axis, showed that education and functional independence were associated with employment. Subjects with a high school education had a 2.2-fold higher chance (95% confidence interval [CI], 1.3 3.8) of returning to work than those without. Subjects with higher scores on the Barthel Index and ability to use public or private transport independently had a 2.7-fold higher chance (95% CI, 1.5-4.9) of returning to work than those unable to travel independently. Other factors significantly associated with employment included marital status, with marriage having a favorable influence; age at injury, with age below 25 years being favorable; preinjury occupation; and vocational training after injury. The importance of functional independence training in rehabilitation is stressed. CONCLUSIONS: Functional independence was a strong factor predicting return to work. Rehabilitation focused on education, vocational training, self-care ability, community mobility, and environmental modifications could improve employability after SCI. PMID- 15827918 TI - Use of orthopedic shoes in patients with degenerative disorders of the foot. AB - OBJECTIVES: To study the actual use of orthopedic shoes by patients with degenerative foot disorders and to identify factors associated with use and nonuse, based on the parameters of the International Organization for Standardization definition of usability: effectiveness, efficiency, satisfaction, and context of use. DESIGN: Multicenter, prospective cohort study. SETTING: Outpatient clinics of 7 rehabilitation centers in the Netherlands. PARTICIPANTS: One hundred consecutive patients with degenerative foot disorders. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Usability was assessed by means of the Questionnaire for Usability Evaluation of orthopedic shoes. RESULTS: Seventy of 93 patients with degenerative foot disorders wore their orthopedic shoes for more than 3 days a week after 3 months of follow-up. Factors significantly associated with the actual use of orthopedic shoes were (1) increase in stance duration (effectiveness odds ratio [OR]=2.14; 95% confidence interval [CI], 1.19-3.85), (2) decrease in skin abnormalities (effectiveness OR=1.35; 95% CI, 1.02-1.8]), (3) problems experienced with putting on and taking off orthopedic shoes (efficiency OR=.46; 95% CI, .26-.82), and (4) cosmetic appearance of orthopedic shoes (satisfaction OR=1.54; 95% CI, 1.1-2.15). The overall fit of the multiple logistic regression model ( R 2 ) was 56.3%. CONCLUSIONS: By adding efficiency and satisfaction factors and not focusing only on the effectiveness factors, the amount of explained variance increases, and it becomes possible to evaluate and design products for people with special needs more comprehensively. PMID- 15827919 TI - Neck pain in demolition derby drivers. AB - OBJECTIVE: To investigate neck pain in demolition derby car drivers. DESIGN: Retrospective survey. SETTING: Internet administered. PARTICIPANTS: Demolition derby drivers who visited the Web site of the International Demolition Derby Association. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Demolition derby neck pain questionnaire. RESULTS: Forty drivers participated in a mean of 30 career events and had an average of 52 car collisions per event, 55% being rear-end. Mean and maximum collision speeds were 26 and 45 miles/h (41.6 km/h, 72 km/h), respectively. Only 2 drivers reported their worst postparticipation neck pain lasted more than 3 months, and for 1 it lasted more than a year; for the majority, the worst neck pain event lasted less than 21 days. Three participants reported having mild chronic persistent derby-related neck pain (never went away). The remaining 37 drivers reported no chronic neck pain. The average pain episode was moderate or severe for 8, but for all respondents, the average pain episodes lasted less than 21 days. Ten reported chronic neck pain they believed was unrelated to derby competition (7 mild, 2 moderate, 1 severe). CONCLUSIONS: These data suggest that derby drivers sustain less chronic neck pain after multiple car collision events than might otherwise be expected. Further study of this unique population of car drivers may contribute to understanding whiplash disorder. PMID- 15827920 TI - Efficacy of dextrose prolotherapy in elite male kicking-sport athletes with chronic groin pain. AB - OBJECTIVE: To determine the efficacy of simple dextrose prolotherapy in elite kicking-sport athletes with chronic groin pain from osteitis pubis and/or adductor tendinopathy. DESIGN: Consecutive case series. SETTING: Orthopedic and trauma institute in Argentina. PARTICIPANTS: Twenty-two rugby and 2 soccer players with chronic groin pain that prevented full sports participation and who were nonresponsive both to therapy and to a graded reintroduction into sports activity. INTERVENTION: Monthly injection of 12.5% dextrose and 0.5% lidocaine into the thigh adductor origins, suprapubic abdominal insertions, and symphysis pubis, depending on palpation tenderness. Injections were given until complete resolution of pain or lack of improvement for 2 consecutive treatments. MAIN OUTCOME MEASURES: Visual analog scale (VAS) for pain with sports and the Nirschl Pain Phase Scale (NPPS), a measure of functional impairment from pain. RESULTS: The final data collection point was 6 to 32 months after treatment (mean, 17 mo). A mean of 2.8 treatments were given. The mean reduction in pain during sports, as measured by the VAS, improved from 6.3+/-1.4 to 1.0+/-2.4 ( P <.001), and the mean reduction in NPPS score improved from 5.3+/-0.7 to 0.8+/-1.9 ( P <.001). Twenty of 24 patients had no pain and 22 of 24 were unrestricted with sports at final data collection. CONCLUSIONS: Dextrose prolotherapy showed marked efficacy for chronic groin pain in this group of elite rugby and soccer athletes. PMID- 15827921 TI - Do older adults with knee osteoarthritis place greater loads on the knee during gait? A preliminary study. AB - OBJECTIVE: To compare the gait of older adults with knee osteoarthritis (OA) to an age-, sex-, and weight-matched healthy cohort that would provide preliminary data to examine the hypothesis that adults with knee OA have abnormal knee joint moments and place greater loads on the knee joint during walking compared with healthy adults. DESIGN: Nonrandomized, descriptive study of healthy and osteoarthritic older adults. SETTING: University clinical research laboratory. PARTICIPANTS: Ten older adults with tibiofemoral and/or patellofemoral radiographic evidence and pain and disability attributed to knee OA and 10 age-, sex-, and weight-matched healthy adults. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Three-dimensional gait analysis to calculate knee joint forces and hip, knee, and ankle joint moments; an analysis of covariance adjusted for differences in walking speed between the groups; electromyographic data to verify our interpretation of the knee joint moment data. RESULTS: The joint forces and moments did not differ statistically between the OA and healthy groups. Nonsignificant differences in the OA group relative to the healthy group included between 7% and 8% greater knee joint compressive (OA group, 3.67+/-0.24 body weight [BW]; healthy group, 3.40+/-0.24 BW) and shear (OA group, 0.47+/-0.04 BW; healthy group, .44+/-.04 BW) forces, 33% higher knee extension moments (OA group, .32+/-.07 Nm/kg; healthy group, .24+/-.07 Nm/kg), and 24% lower knee internal abduction moments (OA group, .25+/-.06 Nm/kg; healthy group, .33+/-.06 Nm/kg). CONCLUSIONS: Previous research suggests that mechanical overload may be associated with knee OA. Our results do not provide statistical evidence to support this hypothesis. Nevertheless, the trends in the data, along with previous results, suggest the need to investigate further the possible existence of a biomechanical pathway to knee OA. PMID- 15827922 TI - Raised paraspinal muscle activity reduces rate of stature recovery after loaded exercise in individuals with chronic low back pain. AB - OBJECTIVES: To further the understanding of stature recovery in subjects with and without chronic low back pain (CLBP) and to determine the relations among stature recovery, paraspinal muscle activity, and perceived pain and disability. DESIGN: A case-control study in which stature loss and recovery were assessed in subjects with and without CLBP after a 20-minute loaded walking task (10% of body mass). Group differences in pain, disability, and paraspinal muscle activity were also assessed. SETTING: A university laboratory. PARTICIPANTS: Twenty subjects with CLBP were matched (age, sex, body mass, physical activity level) with 20 controls who were recruited through notices in the university, general community, and local Primary Care Trust. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Stature changes and integrated electromyograms of the paraspinal muscles during a loading and unloading period were assessed. RESULTS: Stature changes after loading did not differ between groups ( P <.05). Subjects with CLBP recovered significantly less stature during unloading than did the controls ( P <.05). Paraspinal muscle activity correlated negatively with stature recovery ( P <.05). Relations among stature recovery, pain, and disability were shown ( P <.05). CONCLUSIONS: The elevated paraspinal muscle activity exhibited by the CLBP group increased compression on the intervertebral disks and diminished their ability to recover the height lost through loaded exercise. Further research is required to establish whether a change in paraspinal muscle activity is associated with corresponding changes in stature recovery, and if this has any clinical implications by reducing pain and disability. PMID- 15827923 TI - Motor unit synchronization of the vasti muscles in closed and open chain tasks. AB - OBJECTIVES: To investigate motor unit synchronization between medial and lateral vasti and whether such synchronization differs in closed and open chain tasks. DESIGN: Electromyographic recordings of single motor unit action potentials were made from the vastus medialis obliquus (VMO) and multiunit recordings from vastus lateralis during isometric contractions at 30 degrees of knee flexion in closed and open chain conditions. SETTING: Laboratory. PARTICIPANTS: Five volunteers with no history of knee pain (age, 30+/-3.32 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The degree of synchronization between motor unit firing was evaluated by identifying peaks in the electromyographic averages of the vastus lateralis, triggered from motor unit action potentials in the VMO, and the proportion of power in the power spectral density of the triggered average at the firing frequency of the reference motor unit. The proportion of cases in which there was significant power and peaks in the triggered averages was calculated. RESULTS: The proportion of trials with peaks in the triggered averages of the vastus lateralis electromyographic activity was greater than 61.5% in all tasks, and there was a significantly greater proportion of cases where power in the spectrum was greater than 7.5% ( P =.01) for the closed chain condition. CONCLUSIONS: There was a high proportion of synchronized motor units between the 2 muscles during isometric contractions, with evidence for greater common drive between the VMO and vastus lateralis in closed chain tasks. This has implications for rehabilitation because it suggests that closed chain tasks may generate better coordination between the vasti muscles. PMID- 15827924 TI - Back muscle strength and fatigue in healthy and chronic low back pain subjects: a comparative study of 3 assessment protocols. AB - OBJECTIVE: To compare the sensitivity of 3 different back test protocols in measuring differences in strength and fatigue between subjects with and without chronic low back pain (CLBP). DESIGN: Descriptive study using a repeated-measures design. SETTING: A research laboratory within a rehabilitation center. PARTICIPANTS: Eighteen healthy subjects and 13 subjects with CLBP were assessed in a single session to compare the 3 protocols. The protocols were an upright position test (UPP), a semicrouched lifting test (LIF), and the Sorensen fatigue test. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Moments of force and surface electromyography were recorded bilaterally from 4 homologous back muscles while the subjects performed static trunk extension efforts for each protocol. Fatigue was quantified by the slopes of the linear regression of electromyography time-series. RESULTS: The back muscle fatigue and strength scores did not differ significantly for the 2 subject groups for any of the 3 protocols. The electromyography fatigue indices revealed that the Sorensen fatigue test and UPP produced more fatigue in the back muscles than the LIF. CONCLUSIONS: It was impossible to specify which protocol is more sensitive to low back status because no between-group difference was observed for any of the 3 tests. PMID- 15827925 TI - Pulse oximetry does not reliably detect aspiration on videofluoroscopic swallowing study. AB - OBJECTIVE: To examine the reliability of pulse oximetry for identifying aspiration by comparing it with the videofluoroscopic swallowing study (VFSS). DESIGN: Nonrandomized, prospective, double-blind study. SETTING: VFSS laboratory in a teaching hospital. PARTICIPANTS: Sixty patients from among 130 patients with clinically diagnosed dysphagia between September and December 2002. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Simultaneously monitoring the arterial oxygen saturation (Sp o 2 ) by pulse oximetry while patients were performing VFSS. A decrease in Sp o 2 exceeding 3% was considered as significant desaturation. Bolus or portion of bolus passing through the vocal cords and entering the subglottic space was defined as aspiration on VFSS. The results of pulse oximetry and VFSS were compared. RESULTS: No significant correlation existed between desaturation measured by pulse oximetry and aspiration on VFSS (chi 2 test, P =.87). The positive predictive rate of pulse oximetry in detecting aspiration on VFSS was 39.1%, and the negative predictive rate was 59.4%. CONCLUSIONS: Aspiration occurring on VFSS cannot be predicted based on decrease in Sp o 2 in pulse oximetry. The application of pulse oximetry to detect aspiration during regular meals requires further investigation. PMID- 15827926 TI - Community activities and individuals' satisfaction with them: quality of life in the first year after traumatic brain injury. AB - OBJECTIVE: To investigate the relations between community activities and satisfaction with these activities, desires to change them, and global life satisfaction. DESIGN: Interview study with follow-up 1 month after rehabilitation discharge and 12 months postinjury. SETTING: Community. PARTICIPANTS: One hundred sixty-two individuals hospitalized with mostly moderate-to-severe traumatic brain injury. About 90% were reached (mostly by phone) 1 month after rehabilitation discharge; 84.6%, at 12 months postinjury. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Community Integration Questionnaire-2, augmented by individuals' ratings of satisfaction or dissatisfaction with and desire to change each activity; and the Satisfaction With Life Scale. RESULTS: Although significant correlations were found for some items, most correlations between activities and ratings of global quality of life (QOL) were low and nonsignificant. Individuals reported that they were satisfied with most of their community activities, but there were exceptions (eg, paid work). Correlations between activity-specific satisfaction and general life satisfaction were generally weak and nonsignificant. Dissatisfaction with an activity correlated strongly with desire to change the activity, but general life satisfaction did not correlate with desire to change activities. CONCLUSIONS: The lack of association between frequency of activities and subjective appraisals of them is a challenge to outcomes measurement and has implications for the targeting of rehabilitative interventions and evaluation of their worth. More research is needed to understand how individualizing functional objectives might maximize the effects of rehabilitation on the QOL of persons served. PMID- 15827927 TI - The minimally conscious state and recovery potential: a follow-up study 2 to 5 years after traumatic brain injury. AB - OBJECTIVE: To document recovery in persons who were in the minimally conscious state (MCS) for at least 1 month after traumatic brain injury (TBI). DESIGN: Patient series. SETTING: Participants who had been discharged from an inpatient rehabilitation unit. PARTICIPANTS: Eighteen people with TBI and their relatives. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Western Neuro Sensory Stimulation Profile, Disability Rating Scale (DRS), FIM instrument, Dementia Rating Scale, and Sydney Psychosocial Reintegration Scale (SPRS). RESULTS: Level of functioning at follow-up varied from extremely severe disability or greater on the DRS (n=4) to mild disability (n=1). These outcomes were corroborated by results of the FIM and the Dementia Rating Scale. All participants experienced some (44%) or major (56%) change in their level of psychosocial functioning on the SPRS compared with their preinjury level. There were no significant correlations between duration of time in the MCS and outcome on FIM, DRS, or SPRS. CONCLUSIONS: These results highlight the heterogeneity of outcome even after a prolonged duration of MCS after TBI. A large proportion of the patients was functionally independent in basic daily activities, although all experienced residual impairments and disabilities. The low correlation coefficients between duration of MCS and the outcome measures suggest that prognostic statements based on length of time a person is in the MCS cannot be made with confidence. PMID- 15827928 TI - Restoration of weight-shifting capacity in patients with postacute stroke: a rehabilitation cohort study. AB - OBJECTIVES: To identify and interrelate recovery characteristics of voluntary weight shifting after stroke and to examine whether the assessment of weight shifting adds information about balance recovery compared with the assessment of quiet standing. DESIGN: Exploratory study using an inception cohort with findings related to reference values from healthy elderly persons. SETTING: Dutch rehabilitation center. PARTICIPANTS: Thirty-six inpatients (mean age, 61.8 y; mean time poststroke, 10 wk) with a first hemispheric intracerebral infarction or hematoma who were admitted to retrain standing balance and walking. INTERVENTION: Individualized therapy. MAIN OUTCOME MEASURES: Center of pressure (COP) displacements were registered during voluntary frontal-plane weight shifting guided by visual COP feedback using a dual-plate force platform. Besides the speed (number of weight shifts) and imprecision (normalized average lateral COP displacement per weight shift), the weight-transfer time asymmetry and the spatiotemporal distribution were determined. Assessments took place as soon as patients could stand unassisted for at least 30 seconds and at 2, 4, 8, and 12 weeks later. RESULTS: During the 12-week training period, the stroke patients increased both their speed (2.3 hits/30 s; 95% confidence interval [CI], 1.1-3.4) and precision (37.7 mm/hit; 95% CI, 10.4-65.0) of weight shifting. Although the speed appeared to stabilize at a suboptimal level after 8 weeks, precision reached normal reference values after 12 weeks. Both older age (>/=65 y) and the presence of visuospatial hemineglect negatively affected weight-shifting speed but not its relative improvement in time. During the training period, a small degree of weight-transfer time asymmetry persisted (mean change, .07; 95% CI, .21 to .36), with an average of 23% slower weight shifts toward the paretic leg, but the spatiotemporal distribution remained symmetrical. The correlations between weight-shifting and quiet-standing control at the end of training were moderate (Spearman rho range, .50-.77). CONCLUSIONS: Even subjects with severe stroke who are selected for inpatient rehabilitation are able to improve their speed and precision of weight shifting by reducing the weight-transfer time toward both legs in a proportionate manner. The observed correlations between weight shifting and quiet standing indicate that the assessment of weight shifting capacity provides unique information about balance recovery after stroke. PMID- 15827929 TI - Unilateral spatial neglect: relation to rehabilitation outcomes in patients with right hemisphere stroke. AB - OBJECTIVE: To examine the relation between left unilateral spatial neglect (USN) and rehabilitation outcomes in patients with right hemisphere stroke. DESIGN: A retrospective analysis of a database of right hemisphere stroke patients. SETTING: Acute inpatient rehabilitation hospital. PARTICIPANTS: Patients (N=175) with a diagnosis of right hemisphere stroke who had undergone a neuropsychologic screening including assessment of USN and depressive symptoms at time of admission to an inpatient rehabilitation program. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Functional status was evaluated with the FIM instrument at admission and discharge. The relationship between USN, depressive symptoms, cognitive functioning, length of stay (LOS), and rate of progress in rehabilitation was examined via univariate (correlational) and multivariate (Cox regression) analyses. RESULTS: Patients with USN had longer LOS and progressed more slowly compared with those without USN. When matched against patients with equally poor functional status at admission, USN patients still had longer admissions and progressed more slowly. CONCLUSIONS: USN is a unique predictor of rehabilitation outcomes in patients with right hemisphere stroke. Identification of those specific functional skill areas most affected by USN may make possible the development of targeted interventions aimed at these key areas. PMID- 15827930 TI - Support-specific modulation of grip force in individuals with hemiparesis. AB - OBJECTIVE: To investigate whether use of auxiliary sensory input will result in modulated grip force. DESIGN: Case-control study. SETTING: Free-standing acute inpatient rehabilitation hospital. PARTICIPANTS: Six people with unilateral hemiparesis due to unilateral stroke and 6 control subjects without neurologic disorders. INTERVENTIONS: Seated subjects lifted and transported the same object under 3 different conditions: with no support, with the target arm positioned on a freely moving skateboard, and with a finger from the subject's contralateral hand lightly touching the wrist of the target arm. MAIN OUTCOME MEASURES: Peak grip force and temporal coupling between the grip force and lift-off of the object. RESULTS: All subjects were able to better regulate grip force when provided with additional sensory input. Light finger touch resulted in decreased grip force, as did skateboard use ( P <.05). Subjects with hemiparesis showed 2 times longer latency between grip-force application and lift-off of the object ( P <.05). CONCLUSIONS: Statistically significant grip-force reduction was noted with both support aids. These findings could have implications in clinical and rehabilitative areas. PMID- 15827931 TI - Functional implications of ipsilesional motor deficits after unilateral stroke. AB - OBJECTIVE: To investigate the functional impact of ipsilesional motor deficits after unilateral stroke and the best predictors of those deficits. DESIGN: Observational cohort. SETTING: Primary care Veterans Affairs and private medical center. PARTICIPANTS: Volunteer right-handed sample; stroke patients with left (LHD) or right hemisphere damage (RHD) a mean of 3.9 to 5.2 years poststroke and able-bodied participants who were tested using their left (LAB) or right hand. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Jebsen-Taylor Hand Function Test (JHFT). RESULTS: Ipsilesional JHFT performance was impaired to the same extent in the LHD and RHD groups. LHD patients with apraxia had poorer scores on the JHFT than LHD patients without apraxia and the LAB group. Regression analyses showed that severity of apraxia was the best predictor of JHFT performance for the LHD group and that right (ipsilesional) motor performance (grip strength, finger tapping) was the best predictor of JHFT performance for the RHD group. CONCLUSIONS: Ipsilesional deficits are present on simulated activities of daily living after LHD or RHD, suggesting that rehabilitation after stroke should include the ipsilesional arm and that ipsilesional limb apraxia is a better predictor of ipsilesional functional motor skills after LHD than aphasia or simple motor skills (grip strength, finger tapping). These findings suggest that limb apraxia should be assessed more routinely after stroke of the left hemisphere. PMID- 15827932 TI - Test-retest reliability and internal consistency of the Quebec-French version of the Survey of Pain Attitudes. AB - OBJECTIVE: To determine the test-retest reliability and internal consistency of the Quebec-French version of the Survey of Pain Attitudes (QAD/F-SOPA). DESIGN: Measurement of test-retest reliability (2-wk interval) and internal consistency. SETTING: Five rehabilitation settings that offer services to chronic pain patients. PARTICIPANTS: Convenience sample of 69 Francophone adults (having either French as the mother tongue or a good mastery of French) with musculoskeletal pain for a minimum of 6 months and stable pain condition during the test-retest interval. INTERVENTIONS: Completion of the QAD/F-SOPA twice within a 2-week interval. MAIN OUTCOME MEASURES: Test-retest reliability (Pearson r , 2-tail paired t test, P <.001) and internal consistency (Cronbach alpha at time 1). RESULTS: Fifty-six subjects completed the QAD/F-SOPA on both occasions. Except for the disability subscale, the r values fell between 0.7 and 0.9 (high correlation). For the paired t test, all subscales (except for control and medication) had a P value greater than .05, confirming their test-retest stability. All subscales showed satisfactory internal consistency estimates (0.7 0.9) except for the harm (.67) and disability (.64) subscales. CONCLUSIONS: Globally, the QAD/F-SOPA has good reliability and validity properties and meets the prerequisites for use for clinical and research purposes. The disability subscale shows weaker properties; further studies would help determine how it could be improved. PMID- 15827933 TI - Validating the Berg Balance Scale for patients with Parkinson's disease: a key to rehabilitation evaluation. AB - OBJECTIVE: To assess the criterion-related validity of the Berg Balance Scale (BBS) in subjects with Parkinson's disease (PD). DESIGN: Prospective, correlational analysis between the BBS and accepted measures of PD motor and functional impairment. SETTING: The federally funded PD research center, an interdisciplinary center of excellence for people with PD within a Veterans Affairs medical center. PARTICIPANTS: Thirty-eight men (average +/- standard deviation, 71.1+/-10.5 y) with confirmed PD. Their initial diagnosis had been made on average 5.8+/-3.6 years earlier. All could stand or walk unassisted and had mild to moderate disability. Patients who could not ambulate without assistive devices were excluded. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Correlational analyses between the BBS and the Unified Parkinson's Disease Rating Scale (UPDRS) motor scale, Modified Hoehn and Yahr Staging (Hoehn and Yahr) Scale, and the Modified Schwab and England Capacity for Daily Living Scale (S&E ADL Scale). RESULTS: BBS score showed significant correlations with indicators of motor functioning, stage of disease, and daily living capacity. BBS score was inversely associated with the UPDRS motor score (-.58, P <.005), Hoehn and Yahr Scale staging (-.45, P <.005), and S&E ADL Scale rating (.55, P <.005). In all 3 correlations, lower scores on the BBS (indicating greater balance deficits) correlated with higher UPDRS scores (indicating greater motoric or functional impairment). CONCLUSIONS: Results support the criterion-related validity of the BBS. Its utility in other balance conditions of older adults has been established. Rehabilitation interventions have been shown to improve the balance deficits associated with PD. Early referral and periodic reassessment is vital to achieving and maintaining improvements. Our research results agree with other published research in suggesting that the BBS may be used as a screening tool and ongoing assessment tool for patients with PD. PMID- 15827934 TI - Utility of a step activity monitor for the measurement of daily ambulatory activity in children. AB - OBJECTIVES: To evaluate the reliability and validity of the StepWatch Activity Monitor (SAM) as a reliable and valid measurement tool for assessing ambulatory activity in able-bodied children and to assess the ambulatory activity of able bodied children. DESIGN: Descriptive study. SETTING: General community. PARTICIPANTS: Ninety-seven able-bodied children, aged 6 to 20 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Anthropometric parameters, calibration of a step activity monitor to ensure accuracy, and 3 days of simultaneous heart rate and step activity monitoring. RESULTS: The SAM had an accuracy of 99.87% compared with the observer-counted steps and was shown to be valid and reliable when compared with heart rate monitoring. The subjects in all age groups (6-10 y, 11-15 y, 16-20 y) spent most of their active time at low step rate but took the fewest steps at this rate. Although the least amount of time was spent at high step rate, it accounted for the most steps. The 6- to 10-year old group took more total steps per day than any of the other groups. Boys spent significantly more time at high step rate than girls in all age groups (mean for boys, 66+/-4 min/d; girls, 47+/-4 min/d). CONCLUSIONS: The SAM is an accurate, valid, and useful tool for measuring continuous, time-based step activity during real-world community activity for children and adolescents. PMID- 15827935 TI - Use of step activity monitoring for continuous physical activity assessment in boys with Duchenne muscular dystrophy. AB - OBJECTIVES: To evaluate the StepWatch Activity Monitor (SAM) as a quantitative measure of community ambulation, to investigate activity patterns and heart rate of ambulatory boys with Duchenne muscular dystrophy (DMD), and to correlate the step activity with measures of body composition and strength. DESIGN: Case control study. SETTING: General community and laboratory. PARTICIPANTS: Sixteen ambulatory boys with DMD and 20 male controls (age range, 5-13 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Laboratory determinations of body composition, knee extension strength, and minute-by-minute step rate and heart rate during 3 days of community activity. RESULTS: During the 3 days of activity, DMD subjects, when compared with controls, (1) had significantly more inactive minutes (1096+/-90 min/d vs 1028+/-85 min/d), (2) took significantly fewer steps and spent fewer minutes at moderate (66+/-31 min/d vs 94+/-30 min/d) and high step rates (43+/-30 min/d vs 72+/-38 min/d), (3) had higher resting heart rate (110+/-12 beats/min vs 94+/-7 beats/min) and lower increase in heart rate with increased step rate, and (4) had lower maximum heart rates (164+/-24 beats/min vs 208+/-16 beats/min). Percentage of body fat and knee extension strength correlated with total step activity in the DMD group but not in the control group. CONCLUSIONS: Step-rate monitoring with the SAM provides useful outcome measures with which to evaluate the activity of ambulatory boys with DMD. Their heart rate did not increase with activity to the same degree as observed in the control group. PMID- 15827936 TI - Distance measure error induced by displacement of the ulnar nerve when the elbow is flexed. AB - OBJECTIVES: To determine the presence of ulnar nerve displacement at the elbow when it is flexed and to determine its effect on distance measurements using the conventional measurement method for nerve conduction studies (NCSs). DESIGN: Comparing the ultrasonography-assisted distance measurement method with the conventional measurement method. SETTING: An electrodiagnostic laboratory at a university hospital. PARTICIPANTS: Seventy-eight elbows of 39 healthy volunteers. INTERVENTIONS: We used high-resolution ultrasonography in real time. Based on sonographic searching, we marked 3 points on the skin through the course of the displaced ulnar nerve when the elbow is flexed: (1) point A, 7 cm above the elbow (from the midpoint between the medial epicondyle tip and olecranon in the postcondylar groove [point M]); (2) point B, 3 cm below the elbow; and (3) point C, the point closest to the medial epicondyle tip. MAIN OUTCOME MEASURES: Distance measurements between points A, B, and C were taken. These values were compared with measurements obtained through conventional measurement methods. RESULTS: Ulnar nerve displacement occurred in 24.3% (19/78) of the elbows; approximately 20.5% (16/78) were subluxation, and 3.8% (3/78) were dislocation. In the ulnar nerve displacement group, the distance between points A and C was 5.84+/-0.33 cm (range, 5.10-6.30 cm), and the distance between points B and C in the flexed position was 3.35+/-0.19 cm (range, 3.10-3.70 cm). When the conventional distance measurement was used, the ulnar nerve conduction velocity across the elbow was overestimated by approximately 5.33+/-2.29 m/s in the ulnar nerve displacement group. CONCLUSIONS: This distance measurement error may be responsible for the decreased sensitivity found in NCSs that test for ulnar neuropathy at the elbow. If the NCS results are normal in patients who have clear symptoms of ulnar neuropathy, the possibility of ulnar nerve displacement at the elbow should be considered, and further investigation with ultrasonography would be beneficial. PMID- 15827937 TI - Blunted heart rate response to upright tilt in people with Down syndrome. AB - OBJECTIVES: To determine whether heart rate and blood pressure responses to upright tilt would be lower in subjects with Down syndrome (DS) than in control subjects with no disabilities. DESIGN: Comparative study. SETTING: University research laboratory. PARTICIPANTS: Nineteen people with DS (mean age, 25.1+/-7.3 y) and 17 control subjects without disabilities (mean age, 28.4+/-5.6 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Heart rate and blood pressure recordings were obtained at rest and during a 2-minute period of passive head up tilt to 80 degrees . RESULTS: Heart rate and blood pressure increased significantly during the first 30 seconds of upright tilt in both groups ( P <.05) and then stabilized for the remainder of the test. The initial heart rate response to head up tilt (first 30s) was significantly higher in controls compared with subjects with DS ( P <.05), whereas the blood pressure response did not differ between groups ( P >.05). Controlling for heart rate reserve showed a blunted heart rate response through the tilt period in subjects with DS. CONCLUSIONS: These data show a blunted heart rate response to upright tilt in people with DS, despite similar changes in blood pressure, consistent with reduced sympathoexcitation and possibly altered baroreceptor function in these people with DS. PMID- 15827938 TI - The effect of heat on tissue extensibility: a comparison of deep and superficial heating. AB - OBJECTIVE: To compare the effects of deep heating (shortwave diathermy [SWD]) and superficial heating (hydrocollator packs) on tissue extensibility. DESIGN: A double-blind, repeated-measures study. Possible effects of sex and intervention order were controlled. SETTING: A clinical laboratory. PARTICIPANTS: Twenty-four subjects with no neurologic or musculoskeletal pathologies affecting their lower limbs. INTERVENTIONS: Three intervention conditions: deep heating (SWD), superficial heating (hot packs), and no heating were applied in preallocated order to each subject at least 36 hours apart. MAIN OUTCOME MEASURES: Ankle dorsiflexion in weight bearing was measured by using an inclinometer to ascertain changes in the extensibility of the calf muscles and associated soft tissues. RESULTS: Deep heating increased the range of ankle dorsiflexion by 1.8 degrees +/ 1.9 degrees . The change in ankle dorsiflexion after superficial and no heating was 0.7 degrees +/-1.5 degrees and -0.1 degrees +/-1.0 degrees , respectively. CONCLUSIONS: Deep heating, in the absence of stretching, increases tissue extensibility more than superficial heating or no heating. Superficial heating is more effective than no heating, but the difference was not statistically significant. PMID- 15827939 TI - Intrarater reliability of the KT1000 arthrometer in determining anterior translation of the glenohumeral joint. AB - OBJECTIVES: To establish intrarater reliability of the KT1000 arthrometer in determining glenohumeral anterior translation and to determine if a difference existed between measurements of glenohumeral anterior translation in 2 testing positions. DESIGN: Intrarater reliability study. SETTING: Academic laboratory. PARTICIPANTS: Convenience sample of 15 unimpaired volunteers (mean age +/- standard deviation, 25+/-4 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects were positioned supine with the shoulder in 20 degrees of abduction and 0 degrees of external rotation (position 1) and 90 degrees of abduction and 90 degrees of external rotation (position 2). The KT1000 was placed on the shoulder with the tibia sensor pad near the joint line and the patella sensor pad over the coracoid process. Testing involved an anteriorly directed force of 67N. Testing procedures were repeated after 1 week. RESULTS: The intraclass correlation coefficients (ICCs) for intrarater reliability for position 1 (ICC=.93; 95% confidence interval [CI], .81-.98) and for position 2 (ICC=.93; 95% CI, .80-.97) were excellent. The degree of anterior translation measured in position 1 was significantly greater than in position 2 ( t =4.79, P <.01). CONCLUSIONS: Use of the KT1000 to measure glenohumeral anterior translation in the 2 testing positions appears to be a relatively simple, reliable method. Because testing position 1 allowed significantly greater anterior translation than testing position 2, the 2 positions should not be used interchangeably. PMID- 15827940 TI - Treatment of cervical dystonia and focal hand dystonia by high cervical continuously infused intrathecal baclofen: a report of 2 cases. AB - We describe 2 patients, one with cervical dystonia (CD) combined with focal hand dystonia (writer's cramp) and another with idiopathic CD, who were unresponsive to oral medications and became resistant to botulinum toxin type A and B injections. Both patients were successfully treated with high cervical (C1-3) continuously infused intrathecal baclofen (ITB). Neck range of motion (ROM) was measured by using a 3-dimensional electromagnetic cervical ROM system. Pain, disability, and severity were assessed by using the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS). The patient with CD and writer's cramp did well on a continuous baclofen dose of 186.1 microg/d. Her total TWSTRS score improved significantly, her electromagnetic measurements showed an increased in total neck flexion and extension, and her handwriting improved. Unfortunately, this patient (a heavy smoker) developed small cell carcinoma of the lung and died 9 months after her pump was placed. Total TWSTRS score and electromagnetic measurements also significantly improved after pump implant in the patient with CD. He continues to do well on a periodic bolus dose using a combination of 50 microg of baclofen and 25 microg of hydromorphone (Dilaudid) every 4 hours. Our findings suggest the potential usefulness of this therapy in other patients with focal dystonia. To our knowledge, this is the first reported successful treatment of CD and CD combined with writer's cramp with high cervical continuously infused ITB. PMID- 15827941 TI - Rehabilitation of children with traumatic brain injury: descriptive analysis of a nationwide sample using the WeeFIM. AB - OBJECTIVE: To describe functional capability at admission and discharge of children with traumatic brain injury (TBI) in rehabilitation settings. DESIGN: Descriptive analysis. SETTING: Inpatient pediatric rehabilitation hospitals in the United States. PARTICIPANTS: Children (N=3815) in 56 pediatric inpatient rehabilitation facilities who were discharged during 1999 to 2001. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Admission and discharge WeeFIM scores. RESULTS: Admission and discharge WeeFIM scores correlated positively with age at admission, time from injury to rehabilitation admission, and length of stay (LOS). Higher admission WeeFIM scores correlated with shorter LOS, shorter time from injury to admission to rehabilitation, and higher discharge WeeFIM scores. CONCLUSIONS: Children with TBI demonstrated significant improvement in functional measures during rehabilitation. Discharge function and LOS correlated with admission severity, with children who had higher functional status and shorter time between injury and rehabilitation care having higher discharge function and shorter LOS. PMID- 15827942 TI - Methods of constraint-induced movement therapy for children with hemiplegic cerebral palsy: development of a child-friendly intervention for improving upper extremity function. AB - We delineate the methodology for constraint-induced movement therapy (CIMT) modified for children with hemiplegic cerebral palsy (CP) and describe important considerations that need to be made when testing this intervention in children. The resulting intervention evolved from piloting and testing it with 38 children with hemiplegic CP who were between the ages of 4 and 14 years. Thirty-seven successfully completed the treatment protocol. The intervention retains the 2 major elements of the adult CIMT (repetitive practice, shaping) and was constructed to be as child-friendly as possible. It involves restraining the noninvolved extremity with a sling and having the child engage in unimanual activities with the involved extremity 6 hours a day for 10 days (60 h). Specific activities are selected by considering joint movements with pronounced deficits and improvement of which interventionists believe have greatest potential. The activities are chosen to elicit repetitive practice and shaping. The intervention is conducted in groups of 2 to 3 children to provide social interaction, modeling, and encouragement. Each child is assigned to an interventionist to maintain at least a 1:1 ratio. CIMT can be modified to be child-friendly while maintaining all practice elements of the adult CIMT. The modified therapy is tolerated by most children. Further modifications will likely be required to hone in on the specific components of the intervention that are most effective before applying them to children who are most likely to benefit. PMID- 15827943 TI - Spasticity research: some common catches. PMID- 15827945 TI - Geriatric pocket guide and geriatricians in the rehabilitation of older patients. PMID- 15827946 TI - Polyhedral vanadium oxide cages: infrared spectra of cluster anions and size induced d electron localization. PMID- 15827947 TI - A dramatic spin-orbit effect observed in the vibrational frequencies of the chloroiodomethane cation. PMID- 15827948 TI - Direct organocatalytic and highly enantio- and diastereoselective Mannich reactions of alpha-substituted alpha-cyanoacetates. PMID- 15827949 TI - Synthesis and characterization of [exo-BH2(Cp*M)2B9H14] (M = Ru, Re), and the conversion of the ruthenaborane into [(Cp*Ru)2B10H16] with an open cluster framework based on a capped truncated tetrahedron. PMID- 15827950 TI - A new embedded process for compartmentalized cell-free protein expression and on line detection in microfluidic devices. PMID- 15827951 TI - Reviews in Angewandte Chemie: influence, language, competition. PMID- 15827954 TI - Strategies for catalytic asymmetric electrophilic alpha halogenation of carbonyl compounds. PMID- 15827956 TI - Proton affinity ladder for uridine and analogs: influence of the hydroxyl group on the sugar ring conformation. AB - A ladder of relative proton affinities (PA) for a series of modified uridines (e.g. araU, ddU, 5BrU, 5BrdU and 5IU) was established from competitive dissociations of proton-bound heterodimers using Cooks and co-workers' kinetic method. The studied heterodimers are constituted of a modified nucleoside and either an amino acid or a nucleoside with known PA value. These non-covalent heterodimers were prepared under electrospray conditions to be selected and dissociated into the ion-trap analyzer. These results allowed our PA ladder of uridine and deoxyuridine analogs substituted at the C-5 position in the uracil ring to be extended. From this scale, it was showed that the substitution of hydrogen atom at the C-2' position in the sugar ring by a hydroxyl group involves a decrease of about 7 kJ mol(-1). The experimental values for U, 5MeU, dU, 5MedU, ddU and araU are consistent with those obtained by DFT calculations (B3P86/6 31+G//B3LYP/6-31G(.)). Several neutral and protonated conformations of these compounds were considered, in particular the ring conformation of furanose and the orientation of the base with respect to the sugar ring. These calculated results showed the influence of sugar substituent on the conformation of the neutral form of theses nucleosides. However, the most stable protonated structure is the same for all the studied nucleosides except for araU, where the position of the anti 2'-OH group imposes a specific conformation. PMID- 15827957 TI - Novel lipid hydroperoxide-derived hemoglobin histidine adducts as biomarkers of oxidative stress. AB - Hemoglobin (Hb) adducts have long been used as dosimeters of exposure to xenobiotics and endogenously formed reactive metabolites. In this study, hemoglobin chains were separated from each other and their prosthetic heme groups and reacted with 4-oxo-2-nonenal, a major breakdown product of lipid hydroperoxides. The adducts were characterized by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF/MS) analysis of the intact proteins and by a combination of liquid chromatography/electrospray ionization/tandem MS (MS/MS) and MALDI-TOF/MS/MS analysis of the tryptic peptides. Covalent modifications were found on both hemoglobin chains. The location was determined to be on H20 of the alpha-hemoglobin chain and on H(63) of the beta-hemoglobin chain. Molecular modeling revealed that these two residues were two most solvent accessible H residues present in intact Hb. The proposed reaction mechanism is based on that described for the reaction of 4-hydroxy-2 nonenal with proteins. Initial nucleophilic Michael addition is followed by hydration of the resulting aldehyde, cyclization, and two sequential dehydration reactions to give stable furan derivatives. This results in the addition of 136 Da from 4-oxo-2-nonenal to give adducts corresponding to (17)VGAH(.) AGEYGAEALER(31) from alpha-hemoglobin and (62)AH(.) GK(65) from beta-hemoglobin. These hemoglobin modifications can potentially serve as biomarkers of lipid hydroperoxide-mediated macromolecule damage and may reflect an indirect measurement of the potential for DNA damage in vivo. PMID- 15827958 TI - Anomeric distinction and oxonium ion formation in acetylated glycosides. AB - Collision-induced dissociation of ammonium-cationized alpha and beta acetyl pyranosidic isomers were studied and stereochemical dependence of the reactivity towards elimination of acetic acid from the anomeric position was found. It is shown that isomers that contain trans diacetyloxy groups at positions 1 and 2 of the pyranoside are more reactive, allowing anomeric distinction according to the relative abundance of the oxocarbenium product ion of this reaction in the spectrum. The higher reactivity of trans isomers is rationalized by neighboring group assistance that is possible only in the trans configuration. DFT calculations indicate that the lesser energetic reaction path occurs in an ammonium-cationized trans diequatorial 2,3-diacetoxy tetrahydropyran that was used as a model in order to study this process theoretically. It is also found that the configuration at position 4 of the carbohydrate plays a major role in the rate of formation and stability of oxocarbenium ions. PMID- 15827959 TI - Human IL-19 regulates immunity through auto-induction of IL-19 and production of IL-10. AB - IL-19 is a novel, recently identified member of the IL-10 family of cytokines. We identified IL-10 as a cytokine that was strongly induced in IL-19-stimulated PBMC. IL-19-induced IL-10 secretion was dose-dependent and could be detected in culture supernatants after 3 h of stimulation. Furthermore, quantitative RT-PCR analysis demonstrated that IL-19 stimulation increased the level of IL-10 mRNA present within cells, suggesting that IL-19 is a transcriptional activator of IL 10. IL-19 was also able to induce its own expression, with IL-10 potently down regulating this IL-19 'auto-induction'. LPS induction of IL-19 expression was also regulated by IL-10, demonstrating that IL-10 is likely an important regulator of human IL-19 induction. Maturation of dendritic cells from human PBMC in the presence of IL-19 resulted in an increase in IL-10 levels within these cells, whereas IL-12 was not affected. These results advance our understanding of the function of this novel cytokine and its regulation within the human immune system, in addition to providing a new insight into the control of the important immunoregulatory cytokine, IL-10. PMID- 15827960 TI - Bone marrow mesenchymal progenitor cells inhibit lymphocyte proliferation by activation of the programmed death 1 pathway. AB - Bone marrow mesenchymal progenitor cells (BMSC) are used for regenerating tissues of mesodermal origin, as well as tissues of different embryological derivation. Experimental evidence shows that BMSC are able to suppress the activation of the immune response by mechanisms that are still not completely understood. Thus far, in vitro studies carried using human or mouse cells indicate that autologous or allogeneic BMSC strongly suppress proliferation of T lymphocytes, triggered by cellular stimuli, nonspecific mitogenic stimuli, or antigenic peptides. Using cell proliferation and blocking assays, we demonstrated that BMSC inhibited the activation of murine splenocytes, T, and B lymphocytes. Direct contact of BMSC and target cells in a cognate fashion determined the inhibition of cell proliferation via engagement of the inhibitory molecule programmed death 1 (PD-1) to its ligands PD-L1 and PD-L2, leading the target cells to modulate the expression of different cytokine receptors and transduction molecules for cytokine signaling. Soluble factors present on supernatants of BMSC cultures were effective in suppressing proliferation of B cells to a mitogenic stimulus. Taken together, these results highlight the complexity of the role of BMSC in regulating the immune response, asserting the possibility of their therapeutic application in transplantation and autoimmune diseases. PMID- 15827961 TI - The C terminus of T cell-specific adapter protein (TSAd) is necessary for TSAd mediated inhibition of Lck activity. AB - T cell-specific adapter protein (TSAd), encoded by the SH2D2A gene, is expressed in activated T cells. The function of TSAd is as yet unknown. We previously showed that TSAd may modulate T cell receptor-triggered signaling events. TSAd contains a Src homology (SH)2 domain, ten tyrosines and a C-terminal proline-rich region. Here, we show that human TSAd interacts with Lck through the Lck SH2 and SH3 domains and is a substrate for Lck. The TSAd C terminus, including the proline-rich region and five tyrosines, is both necessary and sufficient for TSAd interaction with and phosphorylation by Lck. Expression of TSAd in Jurkat TAg cells results in hyperphosphorylation of endogenous Lck on Y394 and to an even larger extent on Y505, resulting in a reduced Y394/Y505 phosphorylation ratio in these cells. Furthermore, full-length TSAd, but not TSAd lacking the C terminus, inhibits the hyperactive Lck Y505F mutant when both are expressed in Jurkat T cells. In contrast, expression of the TSAd C terminus alone is sufficient to inhibit Lck Y505F in phosphorylating its substrates in Jurkat T cells. Our results indicate that the TSAd C terminus is essential for inhibition of Lck activity by TSAd, and suggest a mechanism for how TSAd may inhibit early T cell activation events. PMID- 15827962 TI - Site-specific expression of CD11b and SIRPalpha (CD172a) on dendritic cells: implications for their migration patterns in the gut immune system. AB - Dendritic cells (DC) in the intestinal tract play a major role in directing the mucosal immune system towards tolerance or immunity. We analyzed whether different mucosal DC subsets in pigs have specific functions, localizations, or migration patterns in vivo. Therefore, we collected physiologically migrating DC by pseudo-afferent cannulation of the intestinal duct in eight Gottingen minipigs. Lymph DC were phenotypically and functionally characterized and compared to DC found on histological sections of porcine small intestine and mesenteric lymph nodes (MLN). Four different DC subpopulations were detected. Lamina propria (LP) DC were mainly CD11b(+) signal regulatory protein alpha (SIRPalpha)(+), DC in Peyer's patches were mainly CD11b(-)/SIRPalpha(+) in subepithelial domes and CD11b(-)/SIRPalpha(-) in interfollicular regions, whereas MLN DC were largely CD11b(+)/SIRPalpha(-). Of these four subsets, only the CD11b(+)/SIRPalpha(+) DC and the CD11b(+)/SIRPalpha(-) DC were present in lymph. This suggests that DC migration to MLN largely originates from the LP. Lymph DC expressed high levels of MHC class II and costimulatory molecules and had a low capacity for FITC-dextran uptake, indicating a mature phenotype. However, lymph DC did not induce PBMC proliferation in MLR, and migration was not significantly influenced by mucosal antigen application. PMID- 15827963 TI - Chemokines synergize in the recruitment of circulating neutrophils into inflamed tissue. AB - The innate immune response against micro-organisms is mediated by phagocytes, attracted by chemokines and other G protein-coupled receptor (GPCR) ligands. Originally, we observed increased neutrophil migration by the interaction of inflammatory CXC chemokines such as IL-8/CXCL8 and granulocyte chemotactic protein (GCP)-2/CXCL6 with regakine-1, a CC chemokine constitutively present in plasma. We here demonstrate statistically significant synergy between regakine-1 and the neutrophil attractants C5a or IL-8/CXCL8 in inducing neutrophil shape change and migration under agarose. In addition, regakine-1 attracted human bone marrow granulocytes and enhanced their chemotactic response to IL-8/CXCL8 in a dose-dependent manner. Thus, plasma chemokines may regulate the number of circulating leukocytes under homeostatic conditions and may facilitate extra recruitment of bone marrow neutrophils during inflammation. Indeed, in vivo, regakine-1 provoked a mild neutrophilia in rabbits upon intravenous injection. We also observed that the CC chemokines regakine-1 and monocyte chemotactic protein 3/CCL7 as well as the CXC chemokine stromal cell-derived factor-1alpha/CXCL12 co operated with murine GCP-2 after intraperitoneal co-administration to increase neutrophil influx in mice. These data demonstrate that inducible and constitutive GPCR ligands synergize to enhance inflammation and facilitate a more effective immune response. PMID- 15827964 TI - Split peripheral tolerance: CD40 ligation blocks tolerance induction for CD8 T cells but not for CD4 T cells in response to intestinal antigens. AB - The role of antigen-presenting cells in the balance between immunity and tolerance to intestinal antigens remains poorly understood. In this study, we examined whether CD40 ligation affects the induction of CD4 and CD8 T cell tolerance in response to intestinal antigens. We show that an agonistic anti-CD40 mAb treatment did not block the induction of OVA-specific CD4 T cell tolerance, whereas this approach enabled strong priming of OVA-specific cytotoxic T cells (CTL), preventing CTL tolerance to intestinal antigen. Such CTL priming was independent of CD4 T cell help but required B7 costimulation. Co-administration of anti-CD40 mAb increased the synthesis of IL-2 and the expression of CD25 by CD8 T cells, but neither IL-2 production nor CD25 expression by CD4 T cells was enhanced by anti-CD40 mAb. However, neutralization of TGF-beta together with addition of agonistic anti-CD40 mAb was able to reverse CD4 T cell tolerance. These findings suggest that the induction of tolerance versus immunity against intestinal antigens is determined by the status of the antigen-presenting cells and that signals via CD40 differently regulate the outcome of CD4 and CD8 T cells in vivo. PMID- 15827965 TI - Regulation of CD4+CD25+ regulatory T cell activity: it takes (IL-)two to tango. AB - Although CD4(+)CD25(+) regulatory T cells (Treg) represent a well-characterized population of T cells with in vitro and in vivo suppressive capacity, the basic mechanisms of suppression are still not understood. The constitutive expression of the high-affinity receptor for IL-2 has raised the question about the role of IL-2 in Treg function. Here, we review recent data indicating that IL-2 is not only necessary for the homeostasis of Treg but is also critical for the activation of Treg function. Since Treg do not produce IL-2 by themselves, their capacity to utilize IL-2 secreted by other T cells appears to be an essential component of Treg biology. This indicates that Treg suppressive activity is controlled by interaction with activated target cells via the soluble mediator IL 2. In Treg, IL-2 has been identified as a potent inducer of the immunosuppressive cytokine IL-10, an important mediator of Treg suppression in vivo. The efficient capture of IL-2 by Treg may, under conditions of limited IL-2 supply, cause IL-2 deprivation of responder T cells. This competition can explain some of the currently discussed discrepancies between in vivo and in vitro activity of Treg. PMID- 15827967 TI - Regulation of mast cells' secretory response by co-clustering the Type 1 Fcepsilon receptor with the mast cell function-associated antigen. AB - The mast cell function-associated antigen (MAFA) is a type II membrane glycoprotein first identified on rat mast cells and basophils. Clustering MAFA inhibits these cells' secretory response to the type 1 Fcepsilon receptor (FcepsilonRI) stimulus. To quantitatively characterize this inhibition and its dependence on MAFA-FcepsilonRI co-clustering, we investigated the secretory response of rat mucosal-type mast cells of the RBL 2H3 line carrying an IgE class, 2,4 dinitrophenyl (DNP) specific monoclonal antibody to DNP-conjugated Fab and F(ab')(2) fragments of (1) mouse IgG, and (2) of the MAFA-specific, monoclonal antibody G63. The first reagent clusters FcepsilonRI-IgE complexes into oligomers by reacting with the DNP residues. The DNP conjugated G63 Fab and F(ab')(2) fragments, additionally aggregate MAFA and form FcepsilonRI-IgE-MAFA co clusters. All experiments using these ligands were performed in the absence or presence of an excess of intact mAb G63, which clusters MAFA molecules. Empirical Hill functions were used to relate the secretory response of mast cells to the equilibrium concentrations of FcepsilonRI-IgE or MAFA clusters and co-clusters calculated as function of the employed ligands concentrations. This analysis of the experimental results indicates that co-clustered MAFA molecules have a markedly higher inhibitory capacity than MAFA-clusters alone. The molecular basis of the enhanced inhibition observed upon co-clustering MAFA with the FcepsilonRI is most probably the increased concentration of the inhibitory cell components in the immediate proximity of the activation coupling elements. PMID- 15827966 TI - gp49B1 deficiency is associated with increases in cytokine and chemokine production and severity of proliferative synovitis induced by anti-type II collagen mAb. AB - Mice with a disrupted gp49B gene, which encodes gp49B1 that is expressed on certain hematopoietic cells and has two immunoreceptor tyrosine-based inhibitory motifs (ITIM), exhibit augmented FcepsilonRI-initiated mast cell degranulation and resultant tissue edema. gp49B1-deficient (gp49B(-/-)) mice also exhibit exaggerated lipopolysaccharide (LPS)-induced intravascular neutrophil aggregation leading to cutaneous microangiopathy. To determine whether gp49B(-/-) mice exhibit elevated cytokine and chemokine levels leading to pathologic inflammation, we quantified clinical and morphologic parameters of arthritis and tissue levels of contributory mediators in gp49B(-/-) and gp49B1-sufficient (gp49B(+/+)) mice injected with anti-type II collagen monoclonal antibody (mAb) and LPS. Clinical scores for joint swelling and histological assessments of synovial thickness and cartilage matrix depletion at day 7 were significantly 2.3 to 2.5-fold greater and were more prolonged in gp49B(-/-) mice. At day 5, the amounts of IL-1beta, macrophage inflammatory protein (MIP)-1alpha, and MIP-2 were 2.1-, 2.5-, and 12-fold greater in joint extracts from gp49B(-/-) mice. A significant 2.7-fold more neutrophils infiltrated the synovium of gp49B(-/-) mice at day 7, and neutrophilia persisted with the delayed resolution of the synovitis. mAb-mediated depletion of neutrophils prevented the synovitis in both strains. Thus, gp49B1 counter-regulates the cytokine and chemokine induction and attendant neutrophilia that are all essential for synovitis and cartilage matrix depletion. PMID- 15827968 TI - Determination of zinc to beta-peptide binding constants with electrospray ionization mass spectrometry. AB - We present an improvement of the titration method for binding constant determination with electrospray ionization (ESI) mass spectrometry that is unaffected by differences in ESI response of measured species in solution. The method consists of a calibration and titration, both using an internal standard that allows relative quantitation. This avoids artifacts such as a decrease in overall signal intensity with increasing ligand concentrations, rendering this approach more reliable and meaningful than direct evaluation of ESI peak intensities. We demonstrate the de novo binding constant determination of novel zinc binding beta-peptides, which have been synthesized with the goal of creating secondary structures stabilized by metal complexation. PMID- 15827969 TI - Screening for and validated quantification of phenethylamine-type designer drugs and mescaline in human blood plasma by gas chromatography/mass spectrometry. AB - In recent years, several newer designer drugs of the so-called 2C series such as 2C-D, 2C-E, 2C-P, 2C-B, 2C-I, 2C-T-2, and 2C-T-7 have entered the illicit drug market as recreational drugs. Some fatal intoxications involving 2C-T-7 have been reported. Only scarce data have been published about analyses of these substances in human blood and/or plasma. This paper describes a method for screening and simultaneous quantification of the above-mentioned compounds and their analog mescaline in human blood plasma. The analytes were analyzed by gas chromatography/mass spectrometry in the selected-ion monitoring mode, after mixed mode solid-phase extraction (HCX) and derivatization with heptafluorobutyric anhydride. The method was fully validated according to international guidelines. Validation data for 2C-T-2 and 2C-T-7 were unacceptable. For all other analytes, the method was linear from 5 to 500 microg/L and the data for accuracy (bias) and precision (coefficient of variation) were within the acceptance limits of +/-15% and <15%, respectively (within +/-20% and <20% near the limit of quantification of 5 microg/L). PMID- 15827970 TI - The influence of mixtures of monodentate achiral ligands on the regioselectivity of transition-metal-catalyzed hydroformylation. PMID- 15827971 TI - A molecular silicon cluster with a "naked" vertex atom*. PMID- 15827972 TI - Application of stereocontrolled stepwise [3+2] cycloadditions to the preparation of inhibitors of alpha4beta1-integrin-mediated hepatic melanoma metastasis. PMID- 15827973 TI - Oligomerization and cyclization processes in the nucleation of microporous silicas. PMID- 15827974 TI - Generation of the elusive meta-benzoquinone in the gas phase. PMID- 15827975 TI - Supercritical water as a solvent. AB - Water is not restricted to moderate temperatures and low pressures, but can exist up to very high temperatures, far above its critical point at 647 K. In this supercritical regime, water can be gradually compressed from gas-like to liquid like densities. The resulting dense supercritical states have extraordinary properties which can be tuned by temperature and pressure, and form the basis for innovative technologies. This Review covers the current knowledge of the major properties of supercritical water and its solutions with nonpolar, polar, and ionic compounds, and of the underlying molecular processes. PMID- 15827976 TI - Engineering virus functionalities on colloidal polyelectrolyte lipid composites. PMID- 15827977 TI - Host solids containing nanoscale anion-binding pockets and their use in selective sensing displacement assays. PMID- 15827978 TI - Influence of dT20 and [d(AT)10]2 on cisplatin hydrolysis studied by two dimensional [1H,15N] HMQC NMR spectroscopy. AB - The influence of the presence of DNA on the kinetics of cisplatin (cis [PtCl2(NH3)2]) aquation (replacement of Cl- by H2O) and anation (replacement of H2O by Cl-) involved in the hydrolysis of cisplatin have been determined by two dimensional [1H,15N] HMQC NMR spectroscopy. Single-stranded dT20 and double stranded [d(AT)10]2 oligonucleotides were used as DNA models, avoiding guanines which are known to react rapidly with aquated cisplatin forms. Reactions starting from cis-[PtCl2(15NH3)2], or from a stoichiometric mixture of cis [Pt(15NH3)2(H2O)2]2+ and Cl- (all 0.5 mM Pt(II); in ionic strength, adjusted to 0.095 M or 0.011 M with NaClO4, pH between 3.0 and 4.0) were followed in an NMR tube in both the absence and presence of 0.7 mM dT20 or [d(AT)10]2. In the presence of dT20, we observed a slight and ionic-strength-independent decrease (15-20 %) of the first aquation rate constant, and a more significant decrease of the second anation rate constant. The latter was more important at low ionic strength, and can be explained by efficient condensation of cis [Pt(15NH3)2(H2O)2]2+ on the surface of single-stranded DNA, in a region depleted of chloride anions. At low ionic strength, we observed an additional set of [1H,15N] HMQC spectral signals indicative of an asymmetric species of PtN2O2 coordination, and we assigned them to phosphate-bound monoadducts of cis [Pt(15NH3)2(H2O)2]2+. Double-stranded [d(AT)10]2 slowed down the first aquation step also by approximately 15 %; however, we could not determine the influence on the second hydrolysis step because of a significant background reaction with cis [Pt(NH3)2(H2O)2]2+. PMID- 15827979 TI - A new pyridine-2,6-bis(oxazoline) for efficient and flexible lanthanide-based catalysts of enantioselective reactions with 3-alkenoyl-2-oxazolidinones. AB - A new pyridine-2,6-bis(oxazoline) (4) has been easily synthesised from the reaction of (1S,2S)-2-amino-1-phenylpropane-1,3-diol (1) and dimethyl pyridine 2,6-dicarboximidate (2), followed by TIPS (TIPS=triisopropylsilyl) protection of the 4'-CH2OH group. The catalysts derived from 4 and eight lanthanide(III) triflates have been tested over three reactions involving 3-acryloyl- and 3 crotonoyloxazolidinones (5 a,b): the Diels-Alder (DA) reaction with cyclopentadiene, the 1,3-dipolar cycloaddition with diphenyl nitrone and the Mukaiyama-Michael reaction with 2-trimethylsilyloxyfuran. Several reactions exhibit very good enantioselectivity (ee>90 %), and the opposite enantiomers can be easily obtained simply by changing the cation. This specific feature of the ligand can be appreciated in the DA reaction of 5 a, since the catalyst [Sc(III)4] gives the adduct (2'S)-9 a with 99 % ee, whereas the catalyst [Y(III)4] gives the opposite enantiomer with 95 % ee. A rationale of the enantioselectivity is proposed on the basis of the NMR spectra of La-based complexes involving 4 and 5 as ligands. PMID- 15827980 TI - Rhodium-catalyzed asymmetric aqueous Pauson-Khand-type reaction. AB - An interesting rhodium-catalyzed asymmetric aqueous Pauson-Khand-type reaction was developed. A chiral atropisomeric dipyridyldiphosphane ligand was found to be highly effective in this system. This operationally simple protocol allows both catalyst and reactants to be handled under air without precautions. Various enynes were transformed to the corresponding bicyclic cyclopentenones in good yield and enantiomeric excess (up to 95 % ee). A study of the electronic effects of the enyne substrates revealed a correlation between the electronic properties of the substrates and the ee value obtained in the product of the Pauson-Khand type reaction. A linear free-energy relationship was observed from a Hammett study. PMID- 15827981 TI - Copolymerization of cyclohexene oxide with CO2 by using intramolecular dinuclear zinc catalysts. AB - The intramolecular dinuclear zinc complexes generated in situ from the reaction of multidentate semi-azacrown ether ligands with Et(2)Zn, followed by treatment with an alcohol additive, were found to promote the copolymerization of CO(2) and cyclohexene oxide (CHO) with completely alternating polycarbonate selectivity and high efficiency. With this type of novel initiator, the copolymerization could be accomplished under mild conditions at 1 atm pressure of CO(2), which represents a significant advantage over most catalytic systems developed for this reaction so far. The copolymerization reaction was demonstrated to be a living process as a result of the narrow polydispersities and the linear increase in the molecular weight with conversion of CHO. In addition, the solid-state structure of the dinuclear zinc complex was characterized by X-ray crystal structural analysis and can be considered as a model of the active catalyst. On the basis of the various efforts made to understand the mechanisms of the catalytic reaction, including MALDI-TOF mass analysis of the copolymers' end-groups, the effect of alcohol additives on the catalysis and CO(2) pressure on the conversion of CHO, as well as the kinetic data gained from in situ IR spectroscopy, a plausible catalytic cycle for the present reaction system is outlined. The copolymerization is initiated by the insertion of CO(2) into the Zn--OEt bond to afford a carbonate ester-bridged complex. The dinuclear zinc structure of the catalyst remains intact throughout the copolymerization. The bridged zinc centers may have a synergistic effect on the copolymerization reaction; one zinc center could activate the epoxide through its coordination and the second zinc atom may be responsible for carbonate propagation by nucleophilic attack by the carbonate ester on the back side of the cis-epoxide ring to afford the carbonate. The mechanistic implication of this is particularly important for future research into the design of efficient and practical catalysts for the copolymerization of epoxides with CO(2.). PMID- 15827982 TI - Borate-catalyzed reactions of hydrogen peroxide: kinetics and mechanism of the oxidation of organic sulfides by peroxoborates. AB - The kinetics of the oxidation of substituted phenyl methyl sulfides by hydrogen peroxide in borate/boric acid buffers were investigated as a function of pH, total peroxide concentration, and total boron concentration. Second-order rate constants at 25 degrees C for the reaction of methyl 4-nitrophenyl sulfide and H(2)O(2), monoperoxoborate, HOOB(OH)(3) (-), or diperoxoborate, (HOO)(2)B(OH)(2) (-), are 8.29 x 10(-5), 1.51 x 10(-2) and 1.06 x 10(-2) M(-1) s(-1), respectively. Peroxoboric acid, HOOB(OH)(2), is unreactive. The Hammett rho values for the reactions of a range of substituted phenyl methyl sulfides and hydrogen peroxide, monoperoxoborate or diperoxoborate are -1.50 +/- 0.1, -0.65 +/ 0.07 and -0.48 (two points only), respectively. The rho values for the peroxoborates are of significantly lower magnitude than expected from their reactivity compared to other peroxides. Nevertheless the negative rho values indicate positive charge development on the sulfur atom in the transition state consistent with nucleophilic attack by the organic sulfides on the peroxoborates as with the other peroxides. The kinetic parameters, including the lack of reactivity of peroxoboric acid, are discussed in terms of the differences in the transition state of reactions involving peroxoboron species with respect to those of other peroxides. PMID- 15827983 TI - A new two-phase route to high-quality CdS nanocrystals. AB - A new two-phase route has been developed to synthesize high-quality CdS nanocrystals with a narrow size distribution and a high photoluminescence (PL) quantum yield (QY). In the two-phase system, toluene and water were used as separate solvents for cadmium myristate (CdM2) and thiourea, which served as cadmium source and sulfur source, respectively, and oleic acid (OA) was used as a ligand for stabilizing the nanocrystals. The reactions were completed in the heated autoclaves. The initial Cd/S molar ratio of the precursors and the reaction temperature were found to be factors that affected the growth of nanocrystals. Furthermore, a seeding-growth technique was developed to synthesize CdS nanocrystals of different sizes, which exhibit PL peaks with quite similar full width at half-maximum (FWHM) values compared to those of the initial nanocrystal seeds in all cases. PMID- 15827984 TI - Assembly of heteropoly acid nanoparticles in SBA-15 and its performance as an acid catalyst. AB - Keggin-type 12-tungstophosphoric acid (TPA) nanocrystals have been assembled inside the pores of mesoporous silica through a vacuum impregnation method by using large-pore SBA-15 as a nanoreactor. The product was characterized by Brunauer-Emmet-Teller particle size distribution (BET-PSD), NMR and FT-IR spectroscopy, X-ray diffraction (XRD), tranmsission electron microscopy (TEM), differential thermal analysis (DTA) and FT-IR of adsorbed pyridine. The experimental results illustrate that the TPA nanocrystals are excellent Bronsted acid catalytic materials at room temperature. PMID- 15827985 TI - Structure-property correlations in solid solutions of (CuI)8P12-xAsx, 2.4< or = x < or =6.6. AB - A series of P/As mixed pnicogen phases of composition (CuI)(8)P(12-x)As(x), in which x = 2.4, 4.2, 4.8, 5.4, and 6.6, have been synthesized and characterized by X-ray single crystal and powder diffraction, solid-state NMR spectroscopy, thermal gravimetric analysis, and impedance spectroscopy. These materials are isostructural to (CuI)(8)P(12) and consist of neutral, tubular P/As mixed pnicogen chains associated with Cu(I) and I(-) ions. The As is distributed throughout the pnicogen chains; however, the "roof" sites of the [P8] cage show preferred occupation by As relative to the other sites. Accordingly, the change in cell volume is not a linear function of the As incorporation. Solid-state (31)P NMR spectroscopy of the 40 % As incorporated sample are consistent with the X-ray structural model, with extensive broadening due to (31)P-(75)As coupling and site disorder, and a change in the chemical shifts of the resonances due to the As substitution into the lattice. The degree of copper ion site disorder, probed by single-crystal X-ray diffraction, increases with increasing As content. Although very little change is observed in the copper ionic conductivity of polycrystalline samples, which ranges from 1.8-5.1 x 10(-6) S cm(-1) for (CuI)(8)P(12-x)As(x), x = 0, 4.2, 5.4; a single crystal (x = 4.8) measured along the needle axis has a conductivity of 1.7 x 10(-3) S cm(-1) at 128 degrees C. This represents an order of magnitude improvement in conductivity over (CuI)(8)P(12) at the same temperature. PMID- 15827986 TI - Tuning photoinduced energy- and electron-transfer events in subphthalocyanine phthalocyanine dyads. AB - A series of subphthalocyanine-phthalocyanine dyads has been prepared by means of palladium-catalyzed cross-coupling reactions between a monoalkynylphthalocyanine and different monoiodosubphthalocyanines. Electronic coupling between the two photoactive units is ensured by a rigid and pi-conjugated alkynyl spacer. In addition, the electronic characteristics of the subphthalocyanine moiety were modulated by the introduction of different peripheral substituents. Cyclic and Osteryoung square-wave voltammetry experiments revealed that the reduction potential of this subunit can be decreased by about 400 mV on going from thioether or no substituents to nitro groups. As a consequence, the energy level of the charge-transfer state could be fine-tuned so as to gain control over the fate of the photoexcitation energy in each subunit. The diverse steady-state and time-resolved photophysical techniques employed demonstrated that, when the charge-transfer state lies high in energy, a quantitative singlet-singlet energy transfer mechanism from the excited subphthalocyanine to the phthalocyanine takes place. On the contrary, stabilization of the radical pair by lowering the redox gap between electron donor and acceptor results in a highly efficient photoinduced electron-transfer process, even in solvents of low polarity such as toluene (Phi(ET) approximately 0.9). These features, together with the extraordinary absorptive cross section that these molecular ensembles display across the whole UV/Vis spectrum, make them model candidates for application in situations where broadband light sources are needed. PMID- 15827987 TI - Versatile glycoblotting nanoparticles for high-throughput protein glycomics. AB - We have developed an effective and practical trap-and-release method based on chemoselective ligation of carbohydrates with reactive aminooxyl groups attached to the surface of nanoparticles (referred to as glycoblotting nanoparticles). These glycoblotting nanoparticles were synthesized by UV irradiation of diacetylene-functionalized lipids that contain the aminooxyl group. The glycoblotting nanoparticles captured carbohydrates in aqueous solution under mild conditions and were collected by simple centrifugation. The trapped carbohydrates were effectively released from the nanoparticles under acidic conditions to give pure oligosaccharides. This glycoblotting process reduced the time required for the purification process of carbohydrates to less than 6 h, compared to the several days needed for conventional chromatographic techniques. The oligosaccharides (N-glycan) were released from ovalbumin (glycoprotein) by PNGase F after tryptic digestion. MALDI-TOF mass spectra before purification did not show any significant signals corresponding to N-glycans because these signals were hidden by the large signals of the abundant peptides. However, after purification with the glycoblotting nanoparticles, only signals corresponding to oligosaccharides appeared. We also demonstrated a clear analysis of the oligosaccharides contained in the mice dermis by means of glycoblotting. PMID- 15827988 TI - Excitation-energy migration in self-assembled cyclic zinc(II)-porphyrin arrays: a close mimicry of a natural light-harvesting system. AB - The excitation-energy-hopping (EEH) times within two-dimensional cyclic zinc(II) porphyrin arrays 5 and 6, which were prepared by intermolecular coordination and ring-closing metathesis reaction of olefins, were deduced by modeling the EEH process based on the anisotropy depolarization as well as the exciton-exciton annihilation dynamics. Assuming the number of energy-hopping sites N = 5 and 6, the two different experimental observables, that is, anisotropy depolarization and exciton-excition annihilation times, consistently give the EEH times of 8.0 +/- 0.5 and 5.3 +/- 0.6 ps through the 1,3-phenylene linkages of 5 and 6, respectively. Accordingly, the self-assembled cyclic porphyrin arrays have proven to be well-defined two-dimensional models for natural light-harvesting complexes. PMID- 15827989 TI - Nanotubes from isomeric dibenzoylmethane molecules. AB - Organic nanotubes of various diameters were fabricated from the isomeric molecule dibenzoylmethane (DBM) by using an immersing technique with ordered porous alumina membrane as the template. The ratio of the enol isomers of DBM increased as the diameters of the nanotubes decreased. In addition, although almost no fluorescence could be detected for the DBM monomer, a striking enhancement in the fluorescence emission intensity of the nanotubes was observed as the diameters decreased. This is due to the increased ratio of the enol isomers. PMID- 15827990 TI - Introduction to the special issue on the placebo concept in psychotherapy. AB - As research on the effectiveness of psychotherapy has flourished over the past two decades, scholars have increasingly borrowed various research strategies, including the placebo concept, from medicine. The recent movement toward evidence based treatment in mental health has also highlighted psychological placebos as control conditions against which to define empirically supported psychotherapies. Nevertheless, the application of the placebo concept to psychotherapy is fraught with both conceptual and pragmatic problems. This special issue brings together leading psychotherapy scholars to explore the concept of the placebo to psychotherapy. Historical, theoretical, ethical, and practical issues are discussed from various perspectives, and recommendations are offered to guide future research. PMID- 15827991 TI - Problems with the use of placebo conditions in psychotherapy research, suggested alternatives, and some strategies for the pursuit of the placebo phenomenon. AB - Due to the numerous conceptual, methodological, and ethical problems that are associated with placebo conditions in psychotherapy research, their use should be abandoned, and more powerful therapy outcome designs (dismantling, additive, parametric, and catalytic) that can contribute to basic knowledge through their ability to isolate specific cause-and-effect relationships are recommended. On the other hand, if indeed the placebo effect is a reliable phenomenon, it would be wise to pursue its causal mechanisms, and some research strategies for initiating such pursuit are briefly described. PMID- 15827992 TI - Placebo psychotherapy: synonym or oxymoron? AB - Contrary to some recent claims, the placebo effect is real and in some cases very substantial. Placebo effects can be produced or enhanced by classical conditioning, but consistent with virtually all contemporary conditioning theories, these effects are generally mediated by expectancy. Expectancy can also produce placebo effects that are inconsistent with conditioning history. Although expectancy also plays an important role in psychotherapy outcome, the logic of placebo-controlled trials does not map well onto psychotherapy research. The idea of evaluating the efficacy of psychotherapy by controlling for nonspecific or placebo factors is based on a flawed analogy and should be abandoned. PMID- 15827993 TI - The placebo is powerful: estimating placebo effects in medicine and psychotherapy from randomized clinical trials. AB - The logic of the randomized double-blind placebo control group design is presented, and problems with using the design in psychotherapy are discussed. Placebo effects are estimated by examining clinical trials in medicine and psychotherapy. In medicine, a recent meta-analysis of clinical trials with treatment, placebo, and no treatment arms was conducted (Hrobjartsson & Gotzsche, 2001), and it was concluded that placebos have small or no effects. A re-analysis of those studies, presented here, shows that when disorders are amenable to placebos and the design is adequate to detect the effects, the placebo effect is robust and approaches the treatment effect. For psychological disorders, particularly depression, it has been shown that pill placebos are nearly as effective as active medications whereas psychotherapies are more effective than psychological placebos. However, it is shown that when properly designed, psychological placebos are as effective as accepted psychotherapies. PMID- 15827994 TI - Experimental analysis of specific treatment factors: efficacy and practice implications. AB - Interest in the empirical demonstration of the clinical efficacy of psychosocial treatments has been rekindled by societal concerns over accountability and cost effectiveness of mental health services. The result has been the identification of specific treatments for specific disorders. The prescription of treatment content should be based on the theory of therapeutic action and/or the disorder to which it is applied. The demonstration of specific treatment efficacy requires experimental evidence showing the influence of specific procedures beyond nonspecific factors of treatment. We provide an analysis of these factors and their effects in evaluating the specific efficacy of prescriptive psychosocial treatments. Experimental procedures and designs that test the validity of specific treatments are described and applied to cognitive-behavioral treatments of generalized anxiety disorder. The empirical and professional implications of specific treatment efficacy in evidence-based practice are discussed. PMID- 15827995 TI - Understanding placebo, nocebo, and iatrogenic treatment effects. AB - Placebo and nonplacebo treatments have both positive and negative effects on patient outcomes. To better understand the patterning of treatment effects, three specific interventions will be discussed that are reported to produce more harm than benefit: critical incident stress debriefing, group therapy for adolescents with conduct disorders, and psychotherapy for dissociative identity disorder. In each case, there is an interaction between mechanisms thought to underlie both placebo and specific treatment effects. Mechanisms hypothesized to underlie placebo and nocebo effects include patient expectancy, self-focused attention to symptoms, motivation to change, and sociocultural role-enactment cues. In the three treatments discussed, specific mechanisms interact with nonspecific mechanisms to produce iatrogenic effects. To advance knowledge, it is important both to specify the theory of treatment and its expected outcomes and to put the theory to test. Only with attention to the empirical findings from programmatic research of specific and nonspecific effects and their interaction is it possible to improve the outcomes of treatment beyond the status quo. PMID- 15827996 TI - Early response in psychotherapy: further evidence for the importance of common factors rather than "placebo effects". AB - Evidence is presented demonstrating that placebo control groups benefit more from psychotherapy than no-treatment control groups but less than patients who receive theory-driven treatments. Through a brief review of the results of comparative outcome studies, studies which compare patient outcomes for those seen by trainees or paraprofessionals versus professional therapists and those which show an early response of a sizable portion of patients, a case is made for the powerful effects of common factors in psychotherapy. The early response phenomenon has proposed another challenge to the unique effects of specific psychotherapies and to the wisdom of emphasizing the causative effects of specific techniques in the treatment of specific disorders. PMID- 15827997 TI - Moving from empirically supported treatment lists to practice guidelines in psychotherapy: the role of the placebo concept. AB - The use of placebos is the "gold standard" in studies of investigational drugs, and of other medical procedures as well. Several recent trends have suggested the use of placebos in studies of psychotherapy to isolate effective treatment components, and as a basis for establishing lists of empirically supported treatments. Unlike within the domain of medicine, in which the logic of placebos is relatively straightforward, the concept of placebo as applied to psychotherapy is fraught with both conceptual and practical problems. The evidence-based practice of psychotherapy can best be promoted through the development of practice guidelines, for which psychotherapy placebos are unnecessary. Moreover, even if problems associated with psychological placebos could be overcome, they are not necessary in psychotherapy research. PMID- 15827998 TI - Reducing exposure of clinical research subjects to placebo treatments. AB - The ethics of pill placebo and placebo psychotherapy conditions in clinical research are controversial. Even when not life threatening, mental disorders dramatically diminish the quality of life. Pill-placebo conditions in drug treatment research have been justified on the grounds that a placebo versus standard drug comparison is necessary to test the quality of the study, viz., the assay sensitivity method. The assay sensitivity method of judging study quality, however, results in misclassification of the quality of some studies, leading to bias in effect size estimation in the context of meta-analyses. This bias is of particular concern in relation to studies comparing psychotherapies to psychotropic drugs, which are conducted outside of the Food and Drug Administration (FDA) context. In cases in which control conditions may be justified on grounds other than as essential elements of an assay sensitivity test, statistical methods to reduce the number of study participants exposed to placebo should be strongly considered. Of the methods available, group sequential methods are the most widely used. Group sequential methods involve successive looks at accumulating data, with rules for terminating a trial (or an arm of a trial) early if results are strong enough. PMID- 15827999 TI - Bilateral cortical representation of the trunk midline in human first somatic sensory area. AB - The cortical representation of the trunk zone in the human first somatosensory area was studied with functional magnetic resonance imaging (fMRI) to establish whether the cutaneous regions close to the midline are represented in this area of both hemispheres. Cortical activation foci evoked by unilateral tactile stimulation of ventral trunk regions were detected in the postcentral gyrus of the contralateral hemisphere slightly medial to or just behind the omega-shaped region of the central sulcus and in the anterior bank of the postcentral sulcus. These regions probably correspond to the trunk ventral midline representation zones of areas 3a-3b and 1-2, respectively. Stimulation of cutaneous regions adjacent to the midline evoked activation foci also in the ipsilateral postcentral gyrus in regions symmetrical to those activated in the contralateral hemisphere. These data demonstrate that in humans, as in nonhuman primates, the cutaneous regions adjacent to the trunk midline are represented bilaterally in the first somatic sensory cortex. Whether the ipsilateral activation depends on callosal or extracallosal inputs remains to be elucidated. PMID- 15828000 TI - Transgenic mice carrying a tetracycline-inducible, truncated transforming growth factor beta receptor (TbetaRII). AB - The transforming growth factor-betas (TGFbetas) have multiple roles, making genetic analysis of their functions difficult. We therefore developed transgenic mouse lines to disrupt TGFbeta signaling using a mechanism that is inducible, reversible, and cell-type specific. The transgenic mouse lines carry an EGFP-pBi DeltaTbetaRII construct (PTR). The DeltaTbetaRII element codes for a dominant negative receptor that is known to disrupt TGFbeta signaling. The DeltaTbetaRII has a c-myc tag. The transgene was silent in the PTR mice, with expression of both EGFP and DeltaTbetaRII occurring when the PTR mice were crossed with mice that express the tetracycline transactivator (CMV-tTA). The expression of EGFP was repressed by the addition of doxycycline to the drinking water of the PTRxCMV tTA mice. The PTR mice were then crossed with neuron-specific-tTA mice. Expression of the DeltaTbetaRII transgene in these mice led to an upregulation of native TGFbeta receptor expression, suggesting that neurons can modulate their responsiveness to TGFbetas. PMID- 15828001 TI - Keratin 19 gene drives Cre recombinase expression throughout the early postimplantation mouse embryo. AB - The development of Cre-lox technology has created new opportunities for studying the tissue-specific functions of genes in vivo during development and disease. We analyzed the spatial and temporal activity of Cre recombinase whose coding sequence was inserted into the endogenous locus for keratin 19. Rather than providing epithelial-specific recombination during organogenesis, this K19cre allele allows unexpected recombination in early embryonic development, resulting in recombination of a loxP-flanked allele throughout all tissues of the mouse, but with sparing of the extraembryonic endoderm, including the anterior visceral endoderm. PMID- 15828002 TI - Transgenic mice that express Cre recombinase under control of a skeletal muscle specific promoter from mef2c. AB - Genes expressed in skeletal muscle are often required in other tissues. This is particularly the case for cardiac and smooth muscle, both contractile tissues that share numerous characteristics with skeletal muscle, such that targeted inactivation can lead to embryonic lethality prior to a requirement for gene function in skeletal muscle. Thus, it is essential that conditional inactivation approaches are developed to disrupt genes specifically in skeletal muscle. In this report, we describe a transgenic mouse that expresses Cre recombinase under the control of a skeletal muscle-specific promoter from the mef2c gene. Cre expression in this transgenic line is completely restricted to skeletal muscle from early in development and is present in all skeletal muscles, including those of epaxial and hypaxial origins and in fast and slow fibers. This early skeletal muscle-specific Cre line will be a useful tool to define the function of genes specifically in skeletal muscle. PMID- 15828003 TI - Crystal structure of a calcium-induced dimer of two isoforms of cobra phospholipase A2 at 1.6 A resolution. AB - The calcium-induced formation of a complex between two isoforms of cobra venom phospholipase A2 reveals a novel interplay between the monomer-dimer and activity inactivity transitions. The monodispersed isoforms lack activity in the absence of calcium ions while both molecules gain activity in the presence of calcium ions. At concentrations higher than 10 mg/ml, in the presence of calcium ions, they dimerize and lose activity again. The present study reports the crystal structure of a calcium-induced dimer between two isoforms of cobra phospholipase A2. In the complex, one molecule contains a calcium ion in the calcium binding loop while the second molecule does not possess an intramolecular calcium ion. However, there are two calcium ions per dimer in the structure. The second calcium ion is present at an intermolecular site and that is presumably responsible for the dimerization. The calcium binding loops of the two molecules adopt strikingly different conformations. The so-called calcium binding loop in the calcium-containing molecule adopts a normal conformation as generally observed in other calcium containing phospholipase A(2) enzymes while the conformation of the corresponding loop in the calcium free monomer deviates considerably with the formation of a unique intraloop Gly33 (N)-Cys27 (O) = 2.74 A backbone hydrogen bond. The interactions of Arg31 (B) with Asp49 (A) and absence of calcium ion are responsible for the loss of catalytic activity in molecule A while interactions of Arg2 (B) with Tyr52 (B) inactivate molecule B. PMID- 15828004 TI - Transgenic mice with pancellular enhanced green fluorescent protein expression in primitive hematopoietic cells and all blood cell progeny. AB - Transgenic mice homogeneously expressing enhanced green fluorescence protein (EGFP) in primitive hematopoietic cells and all blood cell progeny, including erythrocytes and platelets, have not been reported. Given previous data indicating H2Kb promoter activity in murine hematopoietic stem cells (HSCs), bone marrow (BM), and lymphocytes, an H2Kb enhancer/promoter EGFP construct was used to generate transgenic mice. These mice demonstrated pancellular EGFP expression in both primitive BM Sca-1+Lin-Kit+ cells and side population (SP) cells. Additionally, all peripheral blood leukocytes subsets, erythrocytes, and platelets uniformly expressed EGFP strongly. Competitive BM transplantation assays established that transgenic H2Kb-EGFP HSCs had activity equivalent to wildtype HSCs in their ability to reconstitute hematopoiesis in lethally irradiated mice. In addition, immunohistochemistry revealed EGFP transgene expression in all tissues examined. This transgenic strain should be a useful reagent for both murine hematopoiesis studies and functional studies of specific cell types from particular tissues. PMID- 15828005 TI - Folding is not required for bilayer insertion: replica exchange simulations of an alpha-helical peptide with an explicit lipid bilayer. AB - We implement the replica exchange molecular dynamics algorithm to study the interactions of a model peptide (WALP-16) with an explicitly represented DPPC membrane bilayer. We observe the spontaneous, unbiased insertion of WALP-16 into the DPPC bilayer and its folding into an alpha-helix with a transbilayer orientation. The free energy surface suggests that the insertion of the peptide into the DPPC bilayer precedes secondary structure formation. Although the peptide has some propensity to form a partially helical structure in the interfacial region of the DPPC/water system, this state is not a productive intermediate but rather an off-pathway trap for WALP-16 insertion. Equilibrium simulations show that the observed insertion/folding pathway mirrors the potential of mean force (PMF). Calculation of the enthalpic and entropic contributions to this PMF show that the surface bound conformation of WALP-16 is significantly lower in energy than other conformations, and that the insertion of WALP-16 into the bilayer without regular secondary structure is enthalpically unfavorable by 5-10 kcal/mol/residue. The observed insertion/folding pathway disagrees with the dominant conceptual model, which is that a surface-bound helix is an obligatory intermediate for the insertion of alpha-helical peptides into lipid bilayers. In our simulations, the observed insertion/folding pathway is favored because of a large (>100 kcal/mol) increase in system entropy that occurs when the unstructured WALP-16 peptide enters the lipid bilayer interior. The insertion/folding pathway that is lowest in free energy depends sensitively on the near cancellation of large enthalpic and entropic terms. This suggests the possibility that intrinsic membrane peptides may have a diversity of insertion/folding behaviors depending on the exact system of peptide and lipid under consideration. PMID- 15828006 TI - Folding pathways for initiator and effector procaspases from computer simulations. AB - The folding pathways of procaspases 3, 7, and 8 have been studied using a Go-like Hamiltonian and molecular dynamics simulations coupled with a parallel tempering scheme. The folding pathways and the overall structures of procaspases 3 and 7 are similar, and are characterized by monomeric as well as dimeric folding intermediates in agreement with the available structural and thermochemical data. The folding pathway of procaspase 8, on the other hand, is characterized by a larger population of monomers and partially folded dimer intermediates, and only a relatively small population of folded dimer species. The most stable structure predicted for procaspase 8 is a dimer, in which the position of the linker is remarkably different from the one observed in procaspases 3 and 7, leading to the fact that all the contacts that stabilize the active site are essentially formed. This novel and unexpected structure provides a rationale for the observed activity of the procaspase 8 dimer, and thus could be highly relevant for the initiation of FAS-mediated apoptosis. PMID- 15828007 TI - Novel pan-neuronal Cre-transgenic line for conditional ablation of genes in the nervous system. AB - Tissue-specific gene ablation is accomplished by combining conventional gene targeting approaches with site-specific recombinases such as the Cre/loxP system. Despite the use of a cardiac-specific rat myosin light chain II promoter, our transgenic line (CRE3) had little or no Cre expression in the heart; however, strong Cre activity was detected in the brain as early as gestation day E11.5. This was determined by several methods including crossing our mouse line with a lacZ indicator line (ROSA26). Transgenic Cre, in this mouse line, mediated DNA recombination of loxP-flanked genes selectively in neurons throughout the gray matter of the brain, cerebellum, spinal cord, as well as retina, dorsal, and sympathetic ganglia. Cre protein was also detected by immunohistochemistry exclusively in neurons, but not in other types of cells or tissues. Thus, our transgenic CRE3 mice provide pan-neuronal expression of CRE for carrying out conditional deletion of genes in neurons and their progenitors. PMID- 15828008 TI - Computation of conformational entropy from protein sequences using the machine learning method--application to the study of the relationship between structural conservation and local structural stability. AB - A complete protein sequence can usually determine a unique conformation; however, the situation is different for shorter subsequences--some of them are able to adopt unique conformations, independent of context; while others assume diverse conformations in different contexts. The conformations of subsequences are determined by the interplay between local and nonlocal interactions. A quantitative measure of such structural conservation or variability will be useful in the understanding of the sequence-structure relationship. In this report, we developed an approach using the support vector machine method to compute the conformational variability directly from sequences, which is referred to as the sequence structural entropy. As a practical application, we studied the relationship between sequence structural entropy and the hydrogen exchange for a set of well-studied proteins. We found that the slowest exchange cores usually comprise amino acids of the lowest sequence structural entropy. Our results indicate that structural conservation is closely related to the local structural stability. This relationship may have interesting implications in the protein folding processes, and may be useful in the study of the sequence-structure relationship. PMID- 15828009 TI - Selective processing of calling songs by auditory interneurons in the female cricket, Gryllus pennsylvanicus: possible roles in behavior. AB - Female crickets (Gryllus pennsylvanicus), caught in the field as nymphs, responded as adults in the laboratory with selective phonotaxis to model calling songs (CSs) that reproduced the dominant carrier frequencies and syllable periods (SPs) characteristic of the male's natural calling song. Extracellular recordings demonstrated two types of auditory interneurons in the female's cervical connectives that were very similar to the AN1 and AN2 neurons previously described in other gryllid species. The AN2 neuron responded to model CSs with a phasically encoded immediate response, and a more tonically encoded prolonged response. AN2's immediate response exhibited SP-dependent decreases (termed decrement) in its responses to sequential syllables of the CS that were greatest to CSs with the shortest SPs and diminished as SPs were lengthened, resulting in an SP-dependent habituation. Picrotoxin application transformed this SP-dependent habituation by AN2 to SP-selective responses in which the degree of decrement was greatest to SPs that were most phonotactically attractive. AN2's prolonged response was most sensitive to 5 kHz CSs and correlated with the carrier frequency tuning for the thresholds of phonotaxis by females. Thus, in females, AN2's immediate (in the presence of picrotoxin) and prolonged responses were selectively tuned to the SPs and carrier frequencies of the male's calls that were most attractive behaviorally. AN1's responses at threshold were also tuned to the dominant carrier frequencies of the male's CS. PMID- 15828010 TI - Ontogeny of the Marmorkrebs (marbled crayfish): a parthenogenetic crayfish with unknown origin and phylogenetic position. AB - Development, growth, and egg production of the Marmorkrebs (marbled crayfish), a crayfish with parthenogenetic reproduction, uncertain geographic origin, and taxonomic position, was studied under laboratory conditions. Length and weight increments strongly depended on temperature being highest at 30 degrees C, and lowest at 15 degrees C. At 25 degrees C, cephalothorax length and weight increased by 17.5 mm and 1700 mg, respectively, in the course of 150 d, whereas at 15 degrees C these parameters increased by only 7 mm and 100 mg during the same period of time. Photoperiod slightly affected growth at 25 degrees C. During growth experiments, mortality was lower at 20 degrees C compared to higher (25 degrees , 30 degrees C) or lower temperatures (15 degrees C), and lower under short-day than under long-day conditions. Females matured early (at an age of 141 255 d, a cephalothorax length of 14-21.5 mm, and a weight of 0.63-2 g) compared to other crayfish species. Reproductive females with a cephalothorax length of between 25-35 mm produced large clutches (up to 416 eggs) and brooding periods varied between 22 and 42 d. In order to establish a staging scheme for Marmorkrebs embryos, embryos were photographed, externally visible ontogenetic events charted, and dissected embryos stained with a nuclear dye. These experiments indicate that their development is virtually identical to that of other crayfish. In conclusion, these results and others show that the Marmorkrebs may be taken as a representative valid model organism for future developmental studies on Crustacea. PMID- 15828011 TI - Ornithine-urea cycle and urea synthesis in African lungfishes, Protopterus aethiopicus and Protopterus annectens, exposed to terrestrial conditions for six days. AB - The objectives of this study were (1) to determine the type of carbamoyl phosphate synthetase (CPS) present, and the compartmentalization of arginase, in the livers of the African lungfishes, Protopterus aethiopicus and Protopterus annectens, and (2) to elucidate if these two lungfishes were capable of increasing the rates of urea synthesis and capacities of the ornithine-urea cycle (OUC) during 6 days of aerial exposure without undergoing aestivation. Like another African lungfish, Protopterus dolloi, reported elsewhere, the CPS activities from the livers of P. aethiopicus and P. annectens had properties similar to that of the marine ray (Taeniura lymma), but dissimilar to that of the mouse (Mus musculus). Hence, they possessed CPS III, and not CPS I as reported previously. CPS III was present exclusively in the liver mitochondria of both lungfishes, but the majority of the arginase activities were present in the cytosolic fractions of their livers. Glutamine synthetase (GS) activity was also detected in the hepatic mitochondria of both specimens. Therefore, our results suggest that the evolution of CPS III to CPS I might not have occurred before the evolution of extant lungfishes as suggested previously, prompting an examination of the current view on the evolution of CPS and OUC in vertebrates. Aerial exposure led to significant decreases in rates of ammonia excretion in P. aethiopicus and P. annectens, but there were no accumulations of ammonia in their tissues. However, urea contents in their tissues increased significantly after 6 days of aerial exposure. The estimated rates of urea synthesis in P. aethiopicus and P. annectens increased 1.2- and 1.47-fold, respectively, which were smaller than that in P. dolloi (8.6-fold) reported elsewhere. In addition, unlike P. dolloi, 6 days of aerial exposure had no significant effects on the hepatic CPS III activities of P. aethiopicus and P. annectens. In contrast, aerial exposure induced relatively greater degrees of reductions in ammonia production in P. aethiopicus (34%) and P. annectens (37%) than P. dolloi (28%) as previously reported. Thus, our results suggest that various species of African lungfishes respond to aerial exposure differently with respect to nitrogen metabolism and excretion, and it can be concluded that P. aethiopicus and P. annectens depended more on reductions in ammonia production than on increases in urea synthesis to ameliorate ammonia toxicity when exposed to terrestrial conditions. PMID- 15828012 TI - "Blood-contacting neurons" in the brain of the Japanese eel Anguilla japonica. AB - To discriminate "blood-contacting neurons" within the brain of the eel, Evans blue (EB) was injected intraperitoneally. After five days, six brain areas were externally stained blue with the dye; the saccus dorsalis (SD), the epiphysis (E), the area postrema (AP), the posterior part of the magnocellular preoptic nucleus (PM), the pituitary (Pit), and the saccus vasculosus (SV). Among the EB positive area, some cells in the PM, the anterior tuberal nucleus (NAT) and the AP were discriminated as the "blood-contacting neurons" histologically, whereas EB-positive neurons were not detected in the SD, the E, the Pit and the SV regions. In the PM, most EB-positive neurons (90 %) were immunoreactive to vasotocin (AVT) antibody, indicating that these neurons are vasotocinergic. The remaining EB-positive neurons (10 %) were not immunoreactive to ANG II and tyrosine hydroxylase (TH) antibodies. Although some neurons in the PM were immunoreactive to ANG II antibody, they were EB-negative. In contrast, almost all EB-positive neurons in the AP showed TH-like immunoreactivity (-lir), indicating that these neurons utilize catecholamine(s) as a neurotransmitter. The EB positive neurons in the NAT were not immunoreactive to AVT, ANG II and TH antibodies, whereas some neurons without EB-staining showed ANG II-lir. Possible roles of these neurons in regulating drinking behavior in eels are discussed. PMID- 15828013 TI - Cross-fostering in Macropus eugenii leads to increased weight but not accelerated gastrointestinal maturation. AB - Stomach and small intestine development was characterized in tammar wallaby (Macropus eugenii) pouch young (PY) using both morphological and immunohistological criteria. At birth, the stomach is undeveloped in comparison to the well-developed intestinal mucosa. The stomach maintains a uniform morphology in both the forestomach and hindstomach regions until the specialization of cardiac and gastric glands are seen at PY170. Parietal cells, found throughout the mucosa are downregulated in the forestomach as cardiac glandular stomach is developing prior to the transition of the offspring to a diet that includes herbage. In the small intestine, mature-type villi are present at birth but the muscularis externa is immature and undergoes significant development around PY120 onwards. We investigated the effects of changes in maternal milk on gut development in the tammar wallaby using a cross fostering approach that provided younger pouch young with older stage milk. Younger PY (average age 67 days postpartum, n = 5) were transferred onto teats vacated by older stage PY (average age 100 days postpartum, n = 6) for 34 days before gut development was assessed. In addition milk analysis was performed before and after fostering events. Cross-fostered PY animals receiving older stage milk were found to be 31% heavier than controls. There was no difference between carbohydrate and protein concentrations however, fostered PY milk had a higher concentration of lipid than that of controls that may have contributed to heavier fostered PY. No difference was found in stomach or small intestine development between these groups using the criteria employed in this study. PMID- 15828014 TI - Effect of graded hypoxic and acidotic stress on contractile force of heart muscle from hypoxia-tolerant and hypoxia-intolerant turtles. AB - Previous studies have shown that isometric contractile force of in vitro cardiac muscle from the anoxia-tolerant painted turtle, Chrysemys picta bellii, decreases when anoxic and when acidotic. This study sought to define the thresholds for these responses in the isolated ventricular strips of the painted turtle and in the anoxia-intolerant softshell turtles, Apalone spinifera. The ventricular strips were exposed to HCO3- Ringer's solution equilibrated at P(O2) 156, 74, 37, 19, and 0 mmHg (45 min at each grade), at both pH 7.0 and at pH 7.8. Strips were also exposed to graded lactic acidosis with intervals between pH 6.8 and pH 7.8 at P(O2) 156 mmHg (softshell) or 37 mmHg (painted). In painted turtle strips at pH 7.8, force remained at control levels until it decreased by 30% at P(O2) 19 mmHg. No further significant decrease occurred at P(O2) 0. In contrast, softshell turtle muscle force did not fall significantly until P(O2) reached 0. When graded hypoxia was imposed at pH 7.0, strips from both species were more sensitive to hypoxia, but the softshell force decreased at a higher P(O2) than the painted turtle (P(O2) 156 mmHg vs. 37 mmHg), its force fell to a lower level at P(O2) 0 (22 % of control vs. 40 % of control), and unlike painted turtle heart muscle, softshell muscle did not recover fully. In summary, these data indicate that ventricular strips of the painted turtle are no more tolerant of hypoxia alone than strips from the softshell turtle, but that when hypoxia is combined with acidosis, the painted turtle heart muscle functions significantly better during the exposure and recovers more fully after exposure. PMID- 15828015 TI - Temporal and spatial expression of TGF-beta2 in chicken somites during early embryonic development. AB - A multifunctional growth and differentiation factor TGF-beta is expressed at various developmental stages, and its principle role may be involvement in organogenesis. The present study was performed to evaluate the temporal and spatial expression of TGF-beta2 mRNA in developing somites of chicken embryos during their early developmental periods. TGF-betas were expressed in various tissues of the whole embryo obtained at stage 26 (5 days of incubation) as revealed by whole-mount in situ hybridization. TGF-beta2 mRNA was predominantly expressed in somites as well as the head, branchial arch, wing buds, and leg buds. TGF-beta2 mRNA first appeared in the rostral somites on E4, and its expression sites expanded to the middle range of somites at stage 26. At stages 29-31 (6-7 days), expression in the rostral somites disappeared, and it appeared in the caudal somites. TGF-beta2 expression was also analyzed in sections of the embryo by in situ hybridization. The expression sites of TGF-beta2 were clearly observed in the myotomal somite tips as well as the neural tube. RT-PCR analysis showed that TGF-beta2 expression was very low in the blastocyte stage embryo and thereafter increased linearly in the whole trunk until stage 26. These data indicate that TGF-beta2 may be a regulatory factor participating in the somitogenesis of chicken embryos. PMID- 15828016 TI - Different somatostatin receptor subtypes are operating in the brain of the teleost fish, Coris julis. AB - Characterization of somatostatinergic (sst) neuronal activity through the application of nonpeptidyl agonists L-779,976 and L-817,818 which are highly specific for the sst receptors (sstr) sstr(2) and sstr(5), respectively, shows for the first time that sstr2, 5-like subtypes are the two major sstr subtypes operating in the brain of the teleost sea wrasse, Coris julis. A somewhat high but heterogeneous distribution pattern (> 30 < 180 fmol/mg wet tissue weight) of neurons expressing sstr2, 5 was reported in the different diencephalic regions plus in mesencephalon and telencephalon while low values were obtained in the cerebellum. Application of the above nonpeptidyl agonists permitted us to identify sstr2-like as the predominant subtype in telencephalic areas such as the entopeduncular nucleus (E) and postcommissural nucleus of the ventral telencephalon (Vp) as well as in hypothalamic and thalamic areas. At the same time high levels of neurons expressing sstr5-like, that greatly overlap those of sstr2-like in the diencephalic areas such as the anteroventral part of the preoptic nucleus (NPOav), the dorsal habenular nucleus (NHd) and the ventrolateral thalamic nucleus (VL), indicate that sstr2-like is very likely not the only sstr subtype acting in this fish brain. The predominance of sstr5-like in other brain areas is confirmed by the high quantities of this subtype in mesencephalic areas such as the torus longitudinalis (TLo). Overall, the discriminately differing densities of neurons expressing both subtypes seem to point to this system as a key molecular basis accounting for the distinct neurophysiological and behavioral sst-dependent activities in Coris julis. PMID- 15828017 TI - Caspase-3/7 inhibition alters cell morphology in mitomycin-C treated chondrocytes. AB - Apoptosis may play a role in osteoarthritis (OA). Apoptosis can proceed via two different pathways depending on the stimulus. However, both pathways converge upon the effector caspases, caspases-3 and -7. In some systems inhibition of caspases-3 and -7 can prevent apoptosis and may therefore have important therapeutic implications. To confirm this, apoptosis was induced in canine chondrocytes with mitomycin-c (MMC), either in the presence or absence of the general caspase inhibitor, Z-VAD FMK, or a specific caspase-3/7 inhibitor. Z-VAD FMK prevented MMC induced cell death. In contrast, inhibition of caspases-3 and 7 in the presence of MMC induced morphological changes that could be described as necrotic-like or paraptotic-like but did not prevent cell death. The addition of an inhibitor of caspase-8 or caspase-9 along with inhibitor of caspase-3/7 was required to reduce cell death. The morphological changes did not occur in the presence of the caspase-3/7 inhibitor alone and could be prevented by addition of Z-VAD FMK. These data lead to the conclusion that, if the apoptotic program cannot be completed, the cells are pushed into a necrotic or other nonapoptotic mode of death which may involve caspase-8 and/or caspase-9. PMID- 15828018 TI - Investigation on the role of cell transcriptional factor Sp1 and HIV-1 TAT protein in PML onset or development. AB - JC virus (JCV) causes progressive multifocal leukoencephalopathy (PML), characterized by multiple areas of demyelination and attendant loss of brain function. PML is often associated with immunodepression and it is significantly frequent in AIDS patients. The viral genome is divided into early and late genes, between which lies a non-coding control region (NCCR) that regulates JCV replication and presents a great genetic variability. The NCCR of JCV archetype (CY strain) is divided into six regions: A-F containing binding sites for cell factors involved in viral transcription. Deletions and enhancements of these binding sites characterize JCV variants, which could promote viral gene expression and could be more suitable for the onset or development of PML. Therefore, we evaluated by means of polymerase chain reaction (PCR) the presence of JCV genome in cerebrospinal fluid (CSF) of HIV positive and negative subjects both with PML and after sequencing, we analyzed the viral variants found focusing on Sp1 binding sites (box B and D) and up-TAR sequence (box C). It is known that Sp1 activates JCV early promoter and can contribute in maintaining methylation free CpG islands in active genes, while up-TAR sequence is important for HIV-1 Tat stimulation of JCV late promoter. Our results showed that in HIV-positive subjects all NCCR structures presented enhancements of up-TAR element, whereas in HIV-negative subjects both Sp1 binding sites were always retained. Therefore, we can support the synergism HIV-1/JCV in CNS and we can hypothesize that both Sp1 binding sites could be important to complete JCV replication cycle in absence of HIV-coinfection. PMID- 15828019 TI - Polyamine depletion inhibits NF-kappaB binding to DNA and interleukin-8 production in human chondrocytes stimulated by tumor necrosis factor-alpha. AB - The activation of the NF-kappaB pathway by pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNFalpha), can be an important contributor for the re-programming of chondrocyte gene expression, thereby making it a therapeutic target in articular diseases. To search for new approaches to limit cartilage damage, we investigated the requirement of polyamines for NF-kappaB activation by TNFalpha in human C-28/I2 chondrocytes, using alpha-difluoromethylornithine (DFMO), a specific polyamine biosynthesis inhibitor. The NF-kappaB pathway was dissected by using pharmacological inhibitors or by expressing a transdominant IkappaBalpha super repressor. Treatment of C-28/I2 chondrocytes with TNFalpha resulted in a rapid enhancement of nuclear localization and DNA binding activity of the p65 NF-kappaB subunit. TNFalpha also increased the level and extracellular release of interleukin-8 (IL-8), a CXC chemokine that can have a role in arthritis, in an NF-kappaB-dependent manner. Pre-treatment of chondrocytes with DFMO, while causing polyamine depletion, significantly reduced NF-kappaB DNA binding activity. Moreover, DFMO also decreased IL-8 production without affecting cellular viability. Restoration of polyamine levels by the co-addition of putrescine circumvented the inhibitory effects of DFMO. Our results show that the intracellular depletion of polyamines inhibits the response of chondrocytes to TNFalpha by interfering with the DNA binding activity of NF-kappaB. This suggests that a pharmacological and/or genetic approach to deplete the polyamine pool in chondrocytes may represent a useful way to reduce NF-kappaB activation by inflammatory cytokines in arthritis without provoking chondrocyte apoptosis. PMID- 15828020 TI - Extracellular acidification enhances DNA binding activity of MafG-FosB heterodimer. AB - Cells are quite sensitive to a change of the extracellular pH and respond to it through detection of the H+/HCO3- level in extracellular fluid. However, little is known about molecular details induced by acidosis, such as intracellular pathways and gene expression. Here we describe properties of gene expression, protein interaction, and DNA binding activity of basic region leucine zipper (bZIP) transcription factor Maf and FosB during extracellular acidification. When cells were incubated with low pH medium, the expressions of small Maf proteins (MafG, MafK, and MafF) and FosB were clearly increased in an extracellular pH dependent manner and expressed transiently with a peak after 1-2 h after stimulation. Immunofluorescence and protein binding studies indicated that MafG was partially co-localized with FosB in the nucleus and MafG can form heterodimers with FosB at extracellular pH 7.40. Moreover, we found that MafG FosB complexes are able to bind to AP-1 consensus sequence, TGACTCA. To investigate whether extracellular acidification influences to dimerization and DNA binding activity of MafG and FosB, extracellular pH of cultured cells was decreased from 7.40 to 6.80. The decrease in extracellular pH led to enhanced dimerization of MafG with FosB leading to augmentation of the DNA binding activity of the heterodimer to AP-1 consensus sequence. Moreover, extracellular acidification induces mRNA expression of matrix metalloproteinase-1, one of the genes that are regulated by AP-1. These results suggest that MafG-FosB complexes are involved in transcriptional regulation in response to extracellular acidification. PMID- 15828021 TI - Platelet-derived growth factor-BB (PDGF-BB) induces differentiation of bone marrow endothelial progenitor cell-derived cell line TR-BME2 into mural cells, and changes the phenotype. AB - Blood vessels are composed of endothelial cells (EC) and mural cells, and the interaction between EC and mural cells is essential for the development and maintenance of the vasculature. EC differentiate from bone marrow-derived endothelial progenitor cells (EPC). Recently, we established a conditionally immortalized bone marrow EPC-derived cell line, TR-BME2, and a brain capillary EC (BCEC) line, TR-BBB, from temperature-sensitive-SV40 T-antigen gene transgenic rats. To understand the function of EPC, it is important to analyze the difference between EPC and mature EC such as BCEC. In this study, we identified EPC-specific genes by means of subtractive hybridization between TR-BME2 and TR BBB. There was no significant difference between TR-BME2 and TR-BBB in the mRNA level of annexin II, which is expressed in EC. In contrast, the mRNA level of smooth muscle cell (SMC) markers such as smooth muscle protein 22 (SM22), calvasculin, and platelet-derived growth factor (PDGF) receptor-beta, was higher in TR-BME2 than in TR-BBB. Moreover, the mRNA level of contractile SMC markers, such as smooth muscle alpha-actin and SM22, was increased in the absence of EC growth factors, such as vascular endothelial growth factor. The mRNA level of synthetic SMC markers, such as matrix Gla protein, was increased by the addition of PDGF-BB. The SMC derived from TR-BME2 showed an altered phenotype, from contractile-type to synthetic-type, when they were cultured in the absence of PDGF-BB. These results show that TR-BME2 cells have higher levels of SMC markers compared with mature EC, and can differentiate into contractile- or synthetic type SMC. PMID- 15828022 TI - Normal rat hepatic stellate cells respond to endotoxin in LBP-independent manner to produce inhibitor(s) of DNA synthesis in hepatocytes. AB - Endotoxin is implicated in the pathology of acute liver failure. The mechanisms of its actions on quiescent hepatic stellate cells (qHSCs) and their implications in hepatocyte injury are incompletely understood. We investigated effects of endotoxin (bacterial lipopolysaccharide; LPS) on qHSCs and subsequently on hepatocytes. After overnight culture following their isolation, qHSCs were incubated with or without endotoxin for 24 h. The cells and the culture supernatant were analyzed for cytokines and nitric oxide (NO) synthesis. The effects of qHSC-conditioned media on hepatocytes were then determined. LPS increased inducible NO synthase expression, stimulated NO synthesis, and inhibited DNA synthesis in qHSCs. qHSC-conditioned medium inhibited DNA synthesis in hepatocytes without affecting NO synthesis, while LPS (1-1,000 ng/ml) conditioned qHSC medium stimulated NO synthesis and caused further inhibition of DNA synthesis and apoptosis. These effects of LPS were more pronounced when qHSCs were incubated with serum, but not with LPS-binding protein (LBP) although CD14 (a receptor for LPS-LBP complex) was found in qHSCs. LPS stimulated the synthesis of TNF-alpha, interleukin (IL)-6, and IL-1beta but not of TGF-beta in qHSCs. Individually or together, L-N(G)-monomethylarginine and antibodies to IL-1beta, IL-6, and TNF-alpha only partly reversed qHSC + LPS-conditioned medium-induced inhibition of DNA synthesis in hepatocytes. These results suggest that the effects of LPS on qHSCs are novel, occurring without the aid of LBP/CD14. They also indicate that other factors, in addition to NO, TGF-beta, TNF-alpha, IL 1beta, and IL-6 are involved in the mechanisms of the growth inhibitory effects of qHSCs on hepatocytes. PMID- 15828023 TI - CpG-oligodeoxynucleotides induce mobilization of hematopoietic progenitor cells into peripheral blood in association with mouse KC (IL-8) production. AB - The immune system of vertebrates detects bacterial DNA as a "danger signal" based on the presence of unmethylated CpG motifs. We examined whether oligodeoxynucleotides (ODNs) with CpG motifs (CpG-ODNs) also induce mobilization of hematopoietic progenitor cells (HPCs). Mice challenged with CpG-ODNs showed an increase in peripheral blood colony-forming units (CFU) with a peak at day 4 after treatment, associated with an increase, starting 30 min after CpG treatment, in serum levels of mouse keratinocyte-derived chemokine (mKC), a functional homolog of human interleukin (IL) 8; production of granulocyte-colony stimulating factor (CSF) was also detected. Mobilization and mKC induction were sequence-specific and dose-dependent occurring even with low doses of CpG-ODNs. Interestingly, intestinal cells were involved in mKC production. HPC mobilization by CpG-ODNs was dependent on peripheral blood mononuclear cells since mobilization was reduced in neutrophil-depleted mice. Moreover, CpG-ODN treatment significantly increased G-CSF mobilizing capacity. Finally, pretreatment with an anti-mKC neutralizing antibody significantly reduced CpG-induced mobilization, further supporting a role for mKC. Thus, bacterial DNA is a "danger signal" not only for immune cells but also for hematopoietic cells, communicating the need for increased hematopoiesis during infections and for the renewal of the immune system. The HPC mobilization activity of CpG-ODNs will need to be considered in the design of treatment regimens for cancer clinical trials using CpG-ODNs in association with chemotherapy. PMID- 15828025 TI - Pharmacogenomics in colorectal carcinomas: future perspectives in personalized therapy. AB - The recent introduction of new drugs such as capecitabine, irinotecan, and oxaliplatinum has greatly improved the clinical outcome of patients with advanced/metastatic colorectal cancer. Nevertheless, some patients may suffer from the adverse drug reactions which will probably be the main cause of chemotherapy failure. The goal of pharmacogenomics is to find correlations between therapeutic responses to drugs and the genetic profiles of patients; the different responses to a particular drug are due, in fact, not only to the specific clinico-pathological features of the patient or to environmental factors, but also to the ethnic origins and the particular individual's genetic profile. Genes which codify for the metabolism enzymes, receptor proteins, or protein targets of chemotherapy agents often present various genetic polymorphisms. The main aim of this review is to provide an overview of the known polymorphisms present in the genes which codify for factors (thymidylate synthase dihydropyrimidine dehydrogenase, uridine diphosphate (UDP)-glucuronosyl transferase 1A1, enzymes implicated in DNA repair) involved in the action mechanisms of the drugs now utilized in chemotherapeutic treatment of colorectal carcinoma, such as fluoropyrimidines, irinotecan, and platinum agents. PMID- 15828024 TI - Involvement of microtubules and Rho pathway in TGF-beta1-induced lung vascular barrier dysfunction. AB - Transforming growth factor-beta1 (TGF-beta1) is a cytokine critically involved in acute lung injury and endothelial cell (EC) barrier dysfunction. We have studied TGF-beta1-mediated signaling pathways and examined a role of microtubule (MT) dynamics in TGF-beta1-induced actin cytoskeletal remodeling and EC barrier dysfunction. TGF-beta1 (0.1-50 ng/ml) induced dose-dependent decrease in transendothelial electrical resistance (TER) in bovine pulmonary ECs, which was linked to increased actin stress fiber formation, myosin light chain (MLC) phosphorylation, EC retraction, and gap formation. Inhibitor of TGF-beta1 receptor kinase RI (5 microM) abolished TGF-beta1-induced TER decline, whereas inhibitor of caspase-3 zVAD (10 microM) was without effect. TGF-beta1-induced EC barrier dysfunction was linked to partial dissolution of peripheral MT meshwork and decreased levels of stable (acetylated) MT pool, whereas MT stabilization by taxol (5 microM) attenuated TGF-beta1-induced barrier dysfunction and actin remodeling. TGF-beta1 induced sustained activation of small GTPase Rho and its effector Rho-kinase; phosphorylation of myosin binding subunit of myosin specific phosphatase; MLC phosphorylation; EC contraction; and gap formation, which was abolished by inhibition of Rho and Rho-kinase, and by MT stabilization with taxol. Finally, elevation of intracellular cAMP induced by forskolin (50 microM) attenuated TGF-beta1-induced barrier dysfunction, MLC phosphorylation, and protected the MT peripheral network. These results suggest a novel role for MT dynamics in the TGF-beta1-mediated Rho regulation, EC barrier dysfunction, and actin remodeling. PMID- 15828026 TI - Functional expression of TRAIL and TRAIL-R2 during human megakaryocytic development. AB - The expression and function of surface TRAIL and TRAIL receptors were investigated in primary megakaryocytic cells, generated in serum-free liquid phase from peripheral human CD34(+) cells. The surface expression of both TRAIL and "death receptor" TRAIL-R2 became detectable starting from the early phase of megakaryocytic differentiation (day 6 of culture) and persisted at later (days10 14) culture times. On the other hand, "death receptor" TRAIL-R1, "decoy receptors" TRAIL-R3, and TRAIL-R4 were barely detectable or undetectable at any time point examined. Addition of recombinant TRAIL at day 6 of culture increased the rate of spontaneous apoptosis of CD34(+)/CD41(dim) megakaryoblasts and it significantly decreased the total output of mature megakaryocytic cells evaluated after additional 4-8 days of culture. Conversely, addition in culture of TRAIL-R2 Fc chimera, which blocked the interaction between endogenous TRAIL and TRAIL-R2 on the surface of cultured megakaryocytic cells, increased the total megakaryocytic cell count. In addition, recombinant TRAIL promoted a small but reproducible increase of maturation in the surviving megakaryocytic cell population, evaluated by both phenotypic analysis and morphology. A similar pro maturation effect was observed when TRAIL was added to bone marrow-derived CD61(+) megakaryocytic cells. Thus, our data suggest a role of TRAIL as a regulator of megakaryocytopoiesis. PMID- 15828027 TI - Clinical and diagnostic comparison of neonatal alloimmune thrombocytopenia to non immune cases of thrombocytopenia. AB - BACKGROUND: Affected patients with neonatal alloimmune thrombocytopenia (AIT) are often severely thrombocytopenic and, if so, may suffer an intracranial hemorrhage (ICH). This study was undertaken to compare the outcome of cases of AIT to cases of neonatal thrombocytopenia shown not to be AIT and to identify clinical features that would facilitate the diagnosis. PROCEDURE: Two hundred twenty two cases of neonatal thrombocytopenia for which serologic testing was obtained by the referring physician were accrued for this study from 11 testing laboratories. The relevant clinical information was pursued. RESULTS: The mean birth platelet count in 110 neonates with AIT was 26,000/mm(3) x 10(9)/L and the rate of ICH was 11% (not all neonates had head sonos). Three criteria distinguished cases of AIT from other causes of neonatal thrombocytopenia (n = 56): (1) severe thrombocytopenia <50,000/mm(3) x 10(9)/L; (2) ICH associated with 1 or more of: a 1-min Apgar score >5, birthweight >2,200 g, grade >1, antenatal occurrence, or signs of bleeding, that is, petechiae, ecchymoses; and (3) no additional, non hemorrhagic neonatal medical problems. CONCLUSIONS: AIT is a unique type of neonatal thrombocytopenia with significant hemorrhagic consequences. Identification of AIT at the bedside should guide institution of appropriate treatment and lead to serologic testing for confirmation. PMID- 15828028 TI - Organization of transcriptional regulatory machinery in osteoclast nuclei: compartmentalization of Runx1. AB - The osteoclast is a highly polarized multinucleated cell that resorbs bone. Using high resolution immunofluorescence microscopy, we demonstrated that all nuclei of an osteoclast are transcriptionally active. Each nucleus within the osteoclast contains punctately organized microenvironments where regulatory complexes that support transcriptional and post-transcriptional control reside. Functional equivalency of osteoclast nuclei is reflected by similar representation of regulatory proteins that support ribosomal RNA synthesis (nucleolin), mRNA transcription (RNA polymerase II, bromouridine triphosphate), processing of gene transcripts (SC35), signal transduction (NF-kappaB), and phenotypic gene expression (Runx1). Our results establish that gene regulatory machinery is architecturally associated and compartmentalized within intranuclear microenvironments of the multiple nuclei of osteoclasts to support physiologically responsive modifications in cellular structural and functional properties. PMID- 15828029 TI - Determination of enantiomeric composition of (-)-(R)-2-tert butyltetrahydroimidazolidin-4-one by polarimetry, 1H NMR, and chiral SFC. AB - Partial resolution of rac-2-tert-butyltetrahydroimidazolidin-4-one was carried out by recrystallization of diastereomeric salts. The enantiomeric composition of enriched samples was estimated by polarimetry, (1)H NMR, and chiral SFC. Enantiomeric composition estimated by polarimetry or by (1)H NMR was directly proportional to that estimated by chiral SFC. The occurrence of solute self association in chloroform was detected through measurements of optical and specific rotation at variable concentration of (-)-(R)-2-tert butyltetrahydroimidazolidin-4-one. Our data suggest that solute self-association in chloroform might be independent of enantiomeric composition. PMID- 15828030 TI - Spindle pole fragmentation due to proteasome inhibition. AB - During interphase, the centrosome concentrates cell stress response molecules, including chaperones and proteasomes, into a proteolytic center. However, whether the centrosome functions as proteolytic center during mitosis is not known. In this study, cultured mammalian cells were treated with the proteasome inhibitor MG 132 and spindle morphology in mitotic cells was characterized in order to address this issue. Proteasome inhibition during mitosis leads to the formation of additional asters that cause the assembly of multipolar spindles. The cause of this phenomenon was investigated by inhibiting microtubule-based transport and protein synthesis. These experimental conditions prevented the formation of supernumerary asters during mitosis. In addition, the expression of dsRed without proteasome inhibition led to the fragmentation of spindle poles. These experiments showed that the formation of extra asters depends on intact microtubule-based transport and protein synthesis. These results suggest that formation of supernumerary asters is due to excessive accumulation of proteins at the spindle poles and consequently fragmentation of the centrosome. Together, this leads to the conclusion that the centrosome functions as proteolytic center during mitosis and proteolytic activity at the spindle poles is necessary for maintaining spindle pole integrity. PMID- 15828031 TI - Direct high-performance liquid chromatographic enantioseparation of beta-lactam stereoisomers. AB - High-performance liquid chromatographic methods were developed for the separation of the enantiomers of 12 beta-lactams. Direct separations were performed on chiral stationary phases (CSPs) containing cellulose-tris-3,5-dimethylphenyl carbamate (Chiralcel OD-RH and OD-H columns), the macrocyclic glycopeptide antibiotic teicoplanin (Chirobiotic T column), or teicoplanin aglycone (Chirobiotic TAG column) as the chiral selector. It was clearly established that, with teicoplanin-based columns, the teicoplanin aglycone was most often responsible for the enantioseparation of the beta-lactams. The difference in enantioselective free energy between the aglycone CSP and the teicoplanin CSP was in the range between 0.02 and 0.97 kJ mol(-1) for these beta-lactam stereoisomer separations. The separations were carried out with high selectivity and resolution, and the method was therefore suitable for monitoring of the enantiomeric excess after chiral synthesis. The Chirobiotic and Chiralcel columns appear to be highly complementary to one another. The best separation of this class of beta-lactam compound could be obtained using the Chirobiotic TAG in the polar-organic mode plus the Chiralcel OD-H in the normal-phase mode. The elution sequence was also determined. PMID- 15828032 TI - Monitoring proton dissociation and solution conformation of chiral ytterbium complexes with near-IR CD. AB - The ytterbium complex [Yb((S)-THP)](3+) ((S)-THP = (1S,4S,7S,10S-tetrakis(2 hydroxypropyl)-1,4,7,10-tetraazacyclododecane) is investigated in solution through NMR, near-IR absorption, and CD spectroscopy. Quantitative analysis of the paramagnetic pseudocontact NMR shift shows Lambda helicity of the ligand cage around the metal. The NIR CD spectrum recorded at acidic pH is found to be very similar to that of [Yb((R)-DOTMA)](-) ((R)-DOTMA = (1R,4R,7R,10R) alpha,alpha',alpha'',alpha'''-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10 tetraacetic acid), which in solution assumes a twisted square antiprism (TSA) conformation. The similarity of the NIR CD spectra is discussed, and it is the first proof of the Lambda(lambda,lambda,lambda,lambda) conformation of [Yb((S) THP)](3+). NIR CD spectra recorded in the pH range of 2-9 allow one to easily follow proton dissociation and to calculate the pK of this equilibrium in water (pK(A) = 6.4 +/- 0.1). This value agrees well with that determined for [Lu((S) THP)](3+) using potentiometric methods. This demonstrates once again that NIR CD spectroscopy is a powerful technique for investigating the solution structure and dynamics of these complexes. PMID- 15828033 TI - Palladium-catalyzed intermolecular alkenylation of indoles by solvent-controlled regioselective C-H functionalization. PMID- 15828034 TI - Hemiporphycene from the expansion of a corrole ring. PMID- 15828035 TI - 1,3-Dipolar cycloadditions of carbonyl ylides to aldimines: a three-component approach to syn-alpha-hydroxy-beta-amino esters. PMID- 15828036 TI - Design of an axially chiral amino acid with a binaphthyl backbone as an organocatalyst for a direct asymmetric aldol reaction. PMID- 15828037 TI - Hypervalent iodine chemistry in synthesis: scope and new directions. AB - The impressive development of hypervalent iodine chemistry in recent years is reflected by the number of publications in this area. Although the synthesis of the first hypervalent iodine compound dates back more than 100 years, the investigation of the reactivities of these compounds and their efficient use as metal-free reagents in organic synthesis is still ongoing. This contribution summarizes recent achievements and highlights key findings and developments that will influence future research and lead to novel applications of hypervalent iodine reagents in synthesis. PMID- 15828038 TI - Symmetry and topology determine the MoV-CN-MnII exchange interactions in high spin molecules. PMID- 15828039 TI - Cobalt(II)-catalyzed cross-coupling of polyfunctional aryl copper reagents with aryl bromides and chlorides. PMID- 15828040 TI - Patterning through controlled submolecular motion: rotaxane-based switches and logic gates that function in solution and polymer films. PMID- 15828041 TI - Electrochemically controlled ion exchange: proton exchange with sodium zeolite Y. PMID- 15828042 TI - Ultra-rapid synthesis of 15O-labeled 2-deoxy-D-glucose for positron emission tomography (PET). PMID- 15828043 TI - Preparation and selective reactions of mixed bimetallic aromatic and heteroaromatic boron-magnesium reagents. PMID- 15828044 TI - Engineering a 2D protein-DNA crystal. PMID- 15828046 TI - Photonic crystals with thermally switchable stop bands fabricated from Se@Ag2Se spherical colloids. PMID- 15828045 TI - C-H activated isomers of [M(AlCp*)5] (M=Fe, Ru). PMID- 15828047 TI - Mixtures of configurationally stable and fluxional atropisomeric monodentate P ligands in asymmetric Rh-catalyzed olefin hydrogenation. PMID- 15828048 TI - Fabrication of chemical microarrays by efficient immobilization of hydrazide linked substances on epoxide-coated glass surfaces. PMID- 15828049 TI - Identification of a genotoxic mechanism for the carcinogenicity of the environmental pollutant and suspected human carcinogen o-anisidine. AB - 2-methoxyaniline (o-anisidine) is an industrial and environmental pollutant and a bladder carcinogen for rodents. The mechanism of its carcinogenicity was investigated with 2 independent methods, 32P-postlabeling and 14C-labeled o anisidine, to show that o-anisidine binds covalently to DNA in vitro after its activation by human hepatic microsomes. We also investigated the capacity of o anisidine to form DNA adducts in vivo. Rats were treated i.p. with o-anisidine (0.15 mg/kg daily for 5 days) and DNA from several organs was analyzed by 32P postlabeling. Two o-anisidine-DNA adducts, identical to those found in DNA incubated with o-anisidine and human microsomes in vitro, were detected in urinary bladder (4.1 adducts per 10(7) nucleotides), the target organ, and, to a lesser extent, in liver, kidney and spleen. These DNA adducts were identified as deoxyguanosine adducts derived from a metabolite of o-anisidine, N-(2 methoxyphenyl)hydroxylamine. This metabolite was identified in incubations with human microsomes. With 9 human hepatic microsomal preparations, we identified the specific CYP catalyzing the formation of the o-anisidine metabolites by correlation studies and by examining the effects of CYP inhibitors. On the basis of these analyses, oxidation of o-anisidine was attributed mainly to CYP2E1. Using recombinant human CYP (in Supersomes) and purified CYPs, the participation of CYP2E1 in o-anisidine oxidation was confirmed. In Supersomes, CYP1A2 was even more efficient in oxidizing o-anisidine than CYP2E1, followed by CYP2B6, 1A1, 2A6, 2D6 and 3A4. The results, the first report on the potential of the human microsomal CYP enzymes to activate o-anisidine, strongly suggest a carcinogenic potential of this rodent carcinogen for humans. PMID- 15828050 TI - Prediction of lymph node metastasis in colorectal carcinoma by expressionof chemokine receptor CCR7. AB - Chemokine receptors are known to regulate homing of lymphocytes into secondary lymphoid organs and may also be involved in the lymphatic spread of solid tumors. Therefore, the assessment of chemokine receptor expression on colorectal carcinomas could potentially improve the prediction of lymph node spread. This is of great importance for the selection of patients for local therapy without the need for concomitant lymphatic dissection. Currently, only 5% of all patients can be selected for this desirable treatment option by established prognosticators. In a retrospective study, expression levels of the chemokine receptors CCR7, CXCR4 and CXCR5 were determined by immunohistochemistry on paraffin-embedded specimens of 99 colorectal carcinomas, which were curatively operated on, comprising all stages of the disease. Receptor expressions (absent vs. positive) from the overall tumor (OT) and from the invasion front (IF) including further prognosticators were correlated with lymph node status by uni- and multivariate analysis. Data were also correlated with synchronous distant metastases and overall survival. Median follow-up was 64 months. In univariate analysis, lymph node status correlated significantly with lymphovascular invasion, the expression of CCR7 IF, CCR7 OT, CXCR4 IF and CXCR4 OT, as well as pT category. Multivariate analysis revealed a significant correlation of lymph node status with lymphovascular invasion and CCR7 IF expression level. Most interestingly, CCR7 IF expression was significantly linked to decreased survival. CCR7 plays an important role in the mechanism of lymph node spread in colorectal carcinoma. Evaluation of the chemokine receptor expression profile seems to be appropriate to improve the selection of patients suited for local treatment and might constitute targets for nonsurgical therapy. PMID- 15828051 TI - Folate receptor-targeted immunotherapy: induction of humoral and cellular immunity against hapten-decorated cancer cells. AB - We previously exploited the frequent overexpression of folate receptors on cancer cells to decorate malignant cell surfaces selectively with folate-hapten conjugates. In antihapten-immunized hosts, this targeted localization of foreign haptens to tumor cells led to rapid accumulation of autologous antihapten IgG, which in turn yielded potent antitumor activity upon stimulation with cytokines (IL-2, IFN-alpha). In an effort to understand the effector mechanisms responsible for tumor regression, we have now investigated the involvement of both humoral and cellular immune components in the tumor destruction process. We report that the dependence of therapeutic efficacy on folate-hapten concentration is bimodal, suggesting that the conjugate must bridge between a cell surface FR and an antihapten IgG in order to mediate killing. Studies with cancer cells in vitro further demonstrate that folate-fluorescein-marked tumor cells are killed primarily by antibody-dependent cellular cytotoxicity and phagocytosis, with no contribution from complement-dependent mechanisms. Investigations of specific immune cell involvement also reveal that asialo-GM1(+)-natural killer cells, macrophages, CD4+ T cells and CD8+ T cells contribute significantly to recognition/removal of the cancer mass, and that elimination of these cell types markedly compromises the therapy. Because the initial antibody-dependent stage of tumor cell killing is shown to lead to a long-term antibody-independent cellular immunity that involves both CD4+ and CD8+ T cells, we propose that F(c) receptor expressing immune cells not only initiate destruction of the IgG-marked tumor cells, but also participate in presentation of endogenous tumor antigens in a manner that leads to long-term cellular immunity. PMID- 15828052 TI - Randomized trial of dietary fiber and Lactobacillus casei administration for prevention of colorectal tumors. AB - The epidemiologic evidence that dietary fiber protects against colorectal cancer is equivocal. No large-scale clinical study of the administration of Lactobacillus casei has been reported. We examined whether dietary fiber and L. casei prevented the occurrence of colorectal tumors. Subjects were 398 men and women presently free from tumor who had had at least 2 colorectal tumors removed. Subjects were randomly assigned to 4 groups administered wheat bran, L. casei, both or neither. The primary end point was the presence or absence of new colorectal tumor(s) diagnosed by colonoscopy after 2 and 4 years. Among 380 subjects who completed the study, 95, 96, 96 and 93 were assigned to the wheat bran, L. casei, both and no treatment groups, respectively. Multivariate adjusted ORs for occurrence of tumors were 1.31 (95% CI 0.87-1.98) in the wheat bran group and 0.76 (0.50-1.15) in the L. casei group compared to the control group. There was a significantly higher number of large tumors after 4 years in the wheat bran group. The occurrence rate of tumors with a grade of moderate atypia or higher was significantly lower in the group administered L. casei. No significant difference in the development of new colorectal tumors was observed with administration of either wheat bran or L. casei. However, our results suggest that L. casei prevented atypia of colorectal tumors. PMID- 15828053 TI - A review of analytical methods for the determination of trichloroethylene and its major metabolites chloral hydrate, trichloroacetic acid and dichloroacetic acid. AB - Trichloroethylene (TCE) and some of its metabolites are potentially carcinogenic compounds that the general population is commonly exposed to in drinking water. Concentrations of TCE, dichloroacetic acid (DCA) and trichloroacetic acid (TCA) given to laboratory animals in cancer bioassays are high, whereas drinking water levels of the compounds are very low. It is not clear whether the trace amounts of TCE, DCA and TCA in drinking water pose a cancer risk to humans. The accuracy of pharmacokinetic studies relies on the analytical method from which blood and tissue concentration data are obtained. Models that extrapolate cancer risks of TCE and its metabolites from laboratory animals to humans, in turn, rely on the results of pharmacokinetic studies. Therefore, it is essential to have reliable analytical methods for the analysis of TCE and its metabolites. This paper reviews the methods currently in the literature for the analysis of TCE, DCA, TCA and, to a lesser extent, chloral hydrate (CH). Additional aspects of analytical methods such as method validation, species preservation and future directions in the analysis of TCE and its metabolites are also discussed. PMID- 15828054 TI - Natural history of hepatopulmonary syndrome: Impact of liver transplantation. AB - Few data exist concerning survival after the diagnosis of hepatopulmonary syndrome (HPS). Although orthotopic liver transplantation (OLT) frequently results in complete resolution of HPS, the relationship between transplantation and survival has not been described. The study rationale was to describe long term survival in patients with HPS. Data were derived from patients diagnosed with HPS at Mayo Clinic (n = 61) between 1985 and 2002, including those undergoing OLT (n = 24) and those who did not (n = 37). A case-control, Kaplan Meier survival analysis between HPS patients and 77 patients without HPS matched for liver disease cause, model for end-stage liver disease (MELD), severity of liver disease by the Child classification, and age was described for OLT and non OLT groups. Patients with HPS had a mean partial pressure of arterial oxygen (PaO(2)) decline of 5.2 + 2.3 mm Hg per year awaiting OLT. For HPS patients, despite similar baseline PaO(2), brain uptake of technetium macroaggregated albumin ((99m)TcMAA), or measures of hepatic dysfunction, 5-year survival associated with OLT was 76% versus 23% who did not undergo transplantation (P < .0001). Comparing those who did not undergo transplantation, HPS patients had worse 5-year survival than matched controls (P = .0003). However, reasons to deny OLT (comorbidity) in the setting of HPS may well have contributed to observed survival differences. Baseline PaO(2) 0.99. Endogenous compounds did not interfere with tricin detection when the detection wavelength was set at 355 nm, the maximum absorbance of tricin. Accuracy and precision were <15% for all concentrations in all matrices except for the precision at the lower limit of quantification (0.5 microg/mL) in mouse plasma, which was 18.4%. Consumption of diet mixed with tricin at 0.05, 0.2 or 0.5% for one week furnished steady-state levels in plasma, liver and small intestine in the 1--3 x 10(-7), 4--22 x 10(-7) and 3--46 x 10(-5) m ranges, respectively. PMID- 15828059 TI - Assessment of the in-vivo stereochemical integrity of aprepitant based on the analysis of human plasma samples via high-performance liquid chromatography with mass spectrometric detection. AB - Achiral and chiral liquid chromatographic methods utilizing mass spectrometric detection were developed to investigate the possibility of inversion of configuration at any or all of the chiral centers of the neurokinin-1 (NK-1) receptor antagonist, aprepitant (5-[[2(R)-[1(R)-(3,5-bistrifluoromethyl phenyl)ethoxy]-3(S)-(4-fluorophenyl)morpholin-4-yl]methyl]-2,4-dihydro [1,2,4]triazol-3-one), in-vivo, following administration of the compound to man. A structure such as aprepitant, that contains three chiral centers, may exist in eight stereochemical forms or, more specifically, as four diastereoisomeric pairs of enantiomers. The four diastereoisomers were separated from each other using a ProntoSil C18 AQ HPLC column (4.6 x 100 mm, 3 microm particles) with a mobile phase composed of acetonitrile--water (47:53, v/v%). Detection was via a single quadrupole mass spectrometer that was connected to the HPLC system via an APCI interface. Analysis of post-dose plasma samples under these conditions indicated that only aprepitant and or its enantiomer were present following oral administration of the drug. Aprepitant and its enantiomer were separated using a Chiralcel OD-H HPLC column with a mobile phase composed of hexane-isopropanol (80:20, v/v%); tandem mass spectrometric detection using an APCI interface was employed. Post-dose plasma samples analyzed using the Chiracel column were found to contain only aprepitant. The results of these experiments confirm that the products of inversion of configuration at any or all of the three chiral centers of aprepitant are not detectable in human plasma samples obtained following the administration of the drug. PMID- 15828060 TI - Possible steps to the emergence of life: the [GADV]-protein world hypothesis. AB - Based on the fact that RNA has not only a genetic function but also a catalytic function, the RNA world theory on the origin of life was first proposed about 20 years ago. The theory assumes that RNA was amplified by self-replication to increase RNA diversity on the primitive earth. Since then, the theory has been widely accepted as the most likely explanation for the emergence of life. In contrast, we reached another hypothesis, the [GADV]-protein world hypothesis, which is based on pseudo-replication of [GADV]-proteins. We reached this hypothesis during studies on the origins of genes and the genetic code, where [G], [A], [D], and [V] refer to Gly, Ala, Asp, and Val, respectively. In this review, possible steps to the emergence of life are discussed from the standpoint of the [GADV]-protein world hypothesis, comparing it in parallel with the RNA world theory. It is also shown that [GADV]-peptides, which were produced by repeated dry-heating cycles and by solid phase peptide synthesis, have catalytic activities, hydrolyzing peptide bonds in a natural protein, bovine serum albumin. These experimental results support the [GADV]-protein world hypothesis for the origin of life. PMID- 15828061 TI - Analysis of neuroactive amino acids from microdialysate samples by fluorescence detection using a modification of the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate method. AB - A sensitive and rapid reversed-phase high-performance liquid chromatographic method using pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) and fluorescence detection is reported. By directly derivatizing microdialysate samples with AQC, an automatic and rapid simultaneous measurement of aspartate, serine, glutamate, glycine and histidine was developed. Excellent linearity (r2 > or = 0.998) was achieved for the standard mixture used for the validation experiments. Within-day and between-day precision was less than 6.2%, and the accuracy ranged from 95 to 105.2% in standards. This method is suitable for single run analysis of a high number of small volume microdialysate samples from rat hippocampus. Amino acids from microdialysate samples were quantified with RSD for reproducibility below 2%, and at approximately 0.1% for retention time. PMID- 15828062 TI - Introduction of the HPLC method for the determination of quinolone residues in various muscle tissues. AB - For use in veterinary sanitary control of foodstuffs and raw materials of animal origin in Slovenia, we developed a routine and confirmation analytical method for determining the residues of enrofloxacin, ciprofloxacin and flumequine in the muscle tissue of cattle, pigs and poultry. For the muscle tissue of freshwater fish, the determination of the flumequine residues was introduced. The results obtained through simultaneous determination of the residues of enrofloxacin, ciprofloxacin and flumequine showed that the values for the examined antibiotics were up to 600 times lower than the prescribed maximum residue levels (MRL). Another advantage of this method is that it covers a wide range of different fluoroquinolones. PMID- 15828063 TI - Non-aqueous capillary electrophoresis for separation and simultaneous determination of fraxin, esculin and esculetin in Cortex fraxini and its medicinal preparations. AB - A non-aqueous capillary electrophoresis method has been developed for the separation and simultaneous determination of fraxin, esculin and esculetin in Cortex fraxini and its preparation for the first time. Optimum separation of the analytes was obtained on a 47 cm x 75 microm i.d. fused-silica capillary using a non-aqueous buffer system of 60 mM sodium cholate, 20 mM ammonium acetate, 20% acetonitrile and 3% acetic acid at 20 kV and 292 K, respectively. The relative standard deviations (RSDs) of the migration times and the peak heights of the three analytes were in the range of 0.23-0.28 and 2.12-2.60%, respectively. Detection limits of fraxin, esculin and esculetin were 0.1557, 0.4073 and 0.5382 microg/mL, respectively. In the tested concentration range, good linear relationships (correlation coefficients 0.9995 for fraxin, 0.9999 for esculin and 0.9992 for esculetin) between peak heights and concentrations of the analytes were observed. This method has been successfully applied to simultaneous determination of the three bioactive components with the recoveries from 90.2 to 109.2% in the five samples. PMID- 15828064 TI - Differential Trk expression in explant and dissociated trigeminal ganglion cell cultures. AB - During embryonic development, expression of neurotrophin receptor tyrosine kinases (Trks) by sensory ganglia is continuously and dynamically regulated. Neurotrophin signaling promotes selective survival and axonal differentiation of sensory neurons. In embryonic day (E) 15 rat trigeminal ganglion (TG), NGF receptor TrkA is expressed by small diameter neurons, NT-3 receptor TrkC and BDNF receptor TrkB are expressed by large diameter neurons. Organotypic explant and dissociated cell cultures of the TG (and dorsal root ganglia) are commonly used to assay neurotrophin effects on developing sensory neurons. In this study, we compared Trk expression in E15 rat TG explant and dissociated cell cultures with or without neurotrophin treatment. Only a subset of TG cells express each of the three Trk receptors in wholemount explant cultures as in vivo conditions. In contrast, all TG neurons co-express all three Trk receptors upon dissociation, regardless of neurotrophin treatment. Neurons cultured in low concentrations of one neurotrophin first, and switched to higher concentrations of another after 1 day, survive and display morphological characteristics of neurons cultured in a mixture of both neurotrophins for 3 days. Our results indicate that wholemount explant cultures of sensory ganglia represent in vivo conditions in terms of Trk expression patterns; whereas dissociation dramatically alters Trk expression by primary sensory neurons. PMID- 15828066 TI - Quantitation and stability of piperacillin and tazobactam in plasma and ultrafiltrate from patients undergoing continuous venovenous hemofiltration by HPLC. AB - Simple and reproducible HPLC methods for the determination of piperacillin and tazobactam have been developed and a complete stability study carried out. The method for piperacillin plasma samples consisted of protein precipitation with methanol using penicillin G as internal standard. No sample preparation was needed for ultrafiltrate samples. Tazobactam sample preparation involved protein precipitation with acetonitrile and the removal of lipids with dichloromethane. Piperacillin separation was performed on a microBondapack C(18) column (300 x 3.9, 10 microm) and tazobactam on a Novapack C(18) column (150 x 3.9, 4 microm) with UV detection set at 229 and 225 nm, respectively. The mobile phase consisted of phosphate buffer-acetonitrile, delivered at 1.5 mL[sol ]min. Calibration curves determination coefficients were >or=0.999 and response factors CV% < 5%. Intra- and inter-assay precision and accuracy of the quality control and limit of quantification were satisfactory. Plasma and ultrafiltrate samples were stable at -20 and -80 degrees C for 2 months and after three freeze-thaw cycles. In the chromatographic rack, tazobactam ultrafiltrate samples were stable for 24 h and plasma samples for 12 h, piperacillin ultrafiltrate samples for 8 h, but plasma samples for only 4 h. Storage of piperacillin samples at 4 degrees C until analysis is recommended. Piperacillin was stable in the presence of tazobactam. PMID- 15828067 TI - Comparison of self-reported occupational exposure with a job exposure matrix in an international community-based study on asthma. AB - BACKGROUND: Self-reports are frequently used to assess occupational exposures in epidemiological studies on asthma, but the validity and influence of asthma status on performance is unclear. METHODS: Data on self-reported exposure to air pollutants were obtained for 16,752 randomly selected working individuals from 40 study centers, and compared to exposures obtained by a job exposure matrix (JEM). The influence of current asthma symptoms or medication was investigated. RESULTS: Specificity of self-reports amounted to 0.83 and 0.87, and sensitivity 0.48 and 0.42 for asthmatics and non-asthmatics, respectively, when compared with the JEM. Self-reported exposure, but not exposure assessed by the JEM, was more prevalent in areas with a higher community prevalence of asthma. CONCLUSIONS: The prevalence of self-reported occupational exposures seems to depend on asthmatic health status at both the individual and the community level. Associations between self-reports and asthma are likely to be biased, especially in pooled analyses combining different areas with varying prevalence rates of asthma. PMID- 15828068 TI - Mesothelioma in household members of asbestos-exposed workers: 32 United States cases since 1990. AB - BACKGROUND: Mesothelioma is significant as an indicator of asbestos exposure, as a continuing major cause of death in those exposed, and as a risk following lesser exposures. One such exposure is living in the household of an asbestos worker, and coming into contact with fibers brought home on his/her body, clothing, etc. METHODS: Law firms throughout the US known for their pursuit of asbestos claims were polled for mesothelioma claims brought on behalf of family members of identifiable asbestos-exposed workers. Cases with any occupational, environmental, or other possible exposure were not included. RESULTS: This study reports 32 household-exposure mesothelioma cases, diagnosed since 1990. Relationships were wife (15), daughter (11), son (3), sister-in law (1), niece (1), and boarder (1). Occupations of the workers included shipyard (13), insulator (7), and other (12). Of the 27 pleural cases, 13 were epithelial, 5 fibrous, 3 biphasic, and 6 not specified; of the 5 peritoneal cases, 4 were epithelial and 1 fibrous. Latency was greater than 40 years in 27 cases; 6 cases were 40-49 years of age and 17 were 60 or older. CONCLUSIONS: Records from law firms were a useful source of information. Mesothelioma resulting from household exposure is a continuing problem. It is more likely to present in the elderly, after latencies of >40 years. PMID- 15828070 TI - Hospitalized nonfatal injuries in the Alaskan construction industry. AB - BACKGROUND: Construction industry workers are exposed to many hazards leading to fatal and nonfatal injuries. Information for nonfatal work-related injury surveillance may be vague and come from a variety of sources. METHODS: The Alaska Trauma Registry (ATR) is used as an injury surveillance tool to focus on hospitalized nonfatal injuries in the Alaskan construction industry. RESULTS: During 1991-1999, 717 workers in the Alaskan construction industry were hospitalized due to occupational injuries, with an average annual injury rate of 0.39 injuries/100 workers. Leading causes of injury included falls (48%) and machinery (15%). Thirty-four percent of the falls were from a building or structure, followed by falls from a ladder (24%). A fractured bone was the most common type of injury (57%). CONCLUSIONS: Information on hospitalized patients from the ATR focuses on the more severe and debilitating injuries, and provides valuable information for prioritizing injury prevention efforts in Alaska. PMID- 15828069 TI - Morbidity and mortality from hazardous materials events in the personal services industry, 1993-2001: a follow-up report from the Hazardous Substances Emergency Events Surveillance (HSEES) system. AB - BACKGROUND: An estimated 8% of acute hazardous materials (HazMat) events that occur annually in the United States involve victims. Little information is available in the literature pinpointing which industries are associated with these acute events. METHODS: Data from the Agency for Toxic Substances and Disease Registry's Hazardous Substances Emergency Events Surveillance (HSEES) system were analyzed to determine which industry categories had the highest proportion of events with victims. These data were collected from 17 state health departments that participated in the HSEES system from 1993 through 2001. RESULTS: During 1993 through 2001, 53,142 HazMat events occurred, of which 51,989 involved actual releases. Of these events with actual releases, 4,324 (8.3%) involved victims. Of the 14 major industrial categories analyzed, personal services-with 1,311 total events, including 468 with victims-had the highest major industrial category proportion of events with victims (35.7%). This high proportion of events with victims was associated mainly with the following three personal services subcategories: private households; laundry, cleaning, and garment services; and hotels and motels. Most of the victims injured in personal services events involved members of the general public. The most frequently reported symptom was respiratory irritation. The causal factors leading to most releases were operator error, followed by deliberate/illegal damage. CONCLUSIONS: Targeting the personal services industries with appropriate prevention strategies may be an effective way to help begin reducing the high proportion of events with victims in this category. PMID- 15828072 TI - Re: Am J Ind Med 44:204-213, 2003. Scrutinizing ACGIH risk assessments: the trichloroethylene case. PMID- 15828071 TI - Lower back problems and occupational risk factors in a South African steel industry. AB - BACKGROUND: The etiology of work-related back disorders is often population specific. The objective of this study was to identify and establish the association of occupational risk factors with the prevalence of low back (LB) problems in a semi-automated South African Steel industry. METHODS: The design entailed an analytical cross-sectional epidemiological study among a group of 366 steel plant workers. Outcome of LB problems was defined using a guided questionnaire and a functional rating index. Exposure to occupational risk factors was determined using self-reported questionnaires. RESULTS: Multivariate logistic regression analyses indicated significant adjusted odds ratios (OR) for twisting and bending (OR 2.81; CI 1.02-7.73); bulky manual handling (5.58; 1.16 26.71); load carriage (7.20; 1.60-32.37); prolonged sitting (2.33; 1.01-5.37); kneeling and squatting (4.62; 1.28-16.60); and working on slippery and uneven surfaces (3.63; 1.20-10.90). CONCLUSIONS: This study supports the current view of a multifactorial etiology in idiopathic LB problems, and emphasizes the importance of multiple intervention strategies in industrial settings. PMID- 15828073 TI - Effects of theatrical smokes and fogs on respiratory health in the entertainment industry. AB - BACKGROUND: Theatrical fogs (glycol or mineral oil aerosols) are widely used in the entertainment industry to create special effects and make lighting visible. METHODS: We studied 101 employees at 19 sites using fogs and measured personal fog exposures, across work shift lung function, and acute and chronic symptoms. Results were also compared to an external control population, studied previously. RESULTS: Chronic work-related wheezing and chest tightness were significantly associated with increased cumulative exposure to fogs (mineral oil and glycols) over the previous 2 years. Acute cough and dry throat were associated with acute exposure to glycol-based fogs; increased acute upper airway symptoms were associated with increased fog aerosol overall. Lung function was significantly lower among those working closest to the fog source. CONCLUSIONS: Mineral oil- and glycol-based fogs are associated with acute and chronic adverse effects on respiratory health among employees. Reducing exposure, through controls, substitution, and elimination where possible, is likely to reduce these effects. PMID- 15828074 TI - Comparison of asbestos exposure assessments by next-of-kin respondents, by an occupational hygienist, and by a job-exposure matrix from the National Occupational Hazard Survey. AB - BACKGROUND: Assessments of occupational exposures in case-control studies of rapidly fatal illnesses often rely on data from next-of-kin respondents, which may be inaccurate. METHODS: Three methods for assessing exposure to asbestos from case-control data on mesothelioma, including next-of-kin assessment, expert assessment, and use of a generic job-exposure matrix (JEM). Interview data [Spirtas et al. (1994): Occup Environ Med 51:804-811] were reviewed to determine exposure status by an occupational hygienist (C.R.) who was unaware of disease status. Exposure odds ratios were calculated using standard methods, and measures of agreement included the kappa statistic and conditional and marginal odds ratios. RESULTS: Expert assessment detected higher proportions of exposed subjects than the next-of-kin respondents or JEM methods. The disease-exposure odds ratios were highest for respondents, perhaps because of recall bias, and lowest for the JEM method. The agreement was highest between the respondent and expert assessments. A combination of respondent's assessment and JEM assessment led to the best prediction of the expert's assessment. Results for spouse respondents were similar to those for other "next-of-kin" respondents. CONCLUSIONS: Expert assessments were the most plausible, but the data indicate that disease associations could also be detected with the other exposure assessment methods. Using some combination of the proxy respondent's assessment and the JEM assessment, one can predict the expert's assessment. A strategy that relied on the respondent's assessment when it was positive and otherwise obtained an expert assessment could reduce costs with little error, compared to expert assessment on all subjects. PMID- 15828075 TI - Effect of job rotation on need for recovery, musculoskeletal complaints, and sick leave due to musculoskeletal complaints: a prospective study among refuse collectors. AB - BACKGROUND: Job rotation might be an effective preventive measure to reduce the prevalence of musculoskeletal complaints, although its effect has not been yet established. The aim of the present study is to evaluate the effect of job rotation in refuse collecting on need for recovery, prevalence of musculoskeletal complaints, and sick leave due to musculoskeletal complaints. METHODS: A 1-year prospective study among refuse collectors was performed, using standardized questionnaires. Job rotation was performed between collecting two-wheeled containers and driving a refuse truck. The experimental groups of rotating refuse collectors at t(0) and t(1) (group R-R) and non-rotating refuse collectors at t(0) and rotating refuse collectors at t(1) (group NR-R) were compared with a reference group of non-rotating refuse collectors at t(0) and t(1) (group NR-NR). RESULTS: The adjusted need for recovery of group R-R was marginally significantly lower than need for recovery of the reference group. Groups R-R and NR-R had a more than two times higher risk for complaints of the low back than the reference group. No other significant results were found. CONCLUSIONS: Job rotation seemed to coincide with a reduced need for recovery and was associated with an increased risk of low back complaints. No effects were found on sick leave due to musculoskeletal complaints. The results might be influenced by the healthy worker selection effect in the reference group and its inverse in the rotating groups. PMID- 15828076 TI - Laceration injuries among workers at meat packing plants. AB - BACKGROUND: Employees in meat packing experience one of the highest occupational laceration injury rates in the US. METHOD: A retrospective study was conducted using OSHA 200 injury and illness logs and First Reports of Injury from two large US meat packing plants from 1998 to 2000. The total workers observed during the study period ranged between 2,449 and 2,682 per year. RESULTS: Laceration injury incidence rates in Plant 1 were 14.0 injuries per 200,000 person hours (per 100 workers per year) in 1998, 11.5 in 1999, and 8.3 in 2000, whereas in Plant 2 the overall incidence rate was 3.7 in 1998, 4.8 in 1999, and 3.0 in 2000. Laceration injury rates in Plant 2 were close to the expected OSHA recordable laceration injury rate in 1999 (3.0 per 100 workers per year), but Plant 1 was considerably higher. Plant 1 had a kill support department, and removed animal hides whereas Plant 2 did not. Handheld non-powered tools were the most common contact objects whereas the slaughter department had the highest number of injuries. Finger injuries from a handheld non-powered tool were the most frequent. CONCLUSIONS: Findings confirm the high rate of injury from laceration in this industry and indicate hazard varies across time into shift, task being performed, and type of tool being used. PMID- 15828077 TI - Awkward work postures: association with occupational gender segregation. AB - BACKGROUND: Segregation of men and women into different jobs is often cited as one of the most plausible explanations for gender differences in exposure and musculoskeletal disorders. METHODS: Direct measurements of sitting, arm, and trunk postures were taken with two different technical instruments on 156 subjects (78 matched pairs of one female and one male worker) over one full workday in diverse labor markets. RESULTS: Exposure differences between workers were strongly associated with vertical occupational segregation (measured as level of status/authority). The results showed that this association was strongest for female-dominated jobs. Workers in female-dominated jobs with a low status/authority experienced longer duration in standing posture (P = 0.001), and higher frequency of arm elevation (P = 0.028 and 0.040 for the dominant and the non-dominant arm, respectively). They also had longer duration of work with bent trunk compared to corresponding workers with high status/authority (P = 0.035). The association was less pronounced for male-dominated jobs, and no such association was found for gender-integrated jobs. CONCLUSION: The findings have implications for prevention as well as for future research. PMID- 15828078 TI - On the bandwagon. CareFirst joins pay-for-performance trend. PMID- 15828079 TI - Peter James Henry Solomon Benenson. PMID- 15828080 TI - Introduction: Addressing the millennial morbidity--the context of community pediatrics. PMID- 15828081 TI - Displacement is permanent for the Sahrawi refugees. PMID- 15828082 TI - Cross-border trade in medicines causes concern in the EU. PMID- 15828083 TI - Refugee health: questions for a new Commissioner. PMID- 15828084 TI - Ireland's smoking ban is an admirable achievement. PMID- 15828085 TI - Photo quiz. A Thai woman with fever and skin lesions. PMID- 15828086 TI - Photo quiz. Chronic foot ulcers in a patient with hepatitis C. PMID- 15828088 TI - [The mode of reproduction and life science--what does diversity in reproductive processes tell in the post-genome era]. PMID- 15828089 TI - Current awareness in prenatal diagnosis. PMID- 15828087 TI - Analysis of chemokine and cytokine expression in patients with HIV and GB virus type C coinfection. AB - BACKGROUND: Plasma levels of several chemokines and cytokines were evaluated in a cohort of 161 human immunodeficiency virus (HIV)positive patients to shed light on a clinically relevant mechanism that would explain the putative beneficial effect of GB virus type C (GBV-C) coinfection. METHODS: Markers for GBV-C infection were assessed in plasma samples. The syncitium-inducing (SI) capacity of isolated virus from each patient was determined in MT-2 cells. Plasma cytokine and chemokine levels were quantified with use of a commercial enzyme-linked immunosorbent assay. RESULTS: GBV-C viremia was found in 44 (27%) of 161 patients, and anti-E2 antibodies were found in 18 (21%) of 87. In contrast to the findings of ex vivo analysis, no statistically significant differences were observed in levels of CCL5, stromal cell-derived factor 1, interleukin-7, and tumor necrosis factor-alpha in plasma of patients with or without GBV-C viremia. Seventy-two (45%) and 89 (55%) of our patients harbored SI and non-SI (NSI) strains, respectively. GBV-C viremia was less prevalent among patients with SI strains (13 [18%] of 72) than among patients with NSI strains (30 [34%] of 89; P = .6). Of interest, coinfected patients with SI strains had significantly higher CD4+ T cell values than did patients who were not coinfected. CONCLUSIONS: Our results suggest that GBV-C infection does not appear to influence the expression of the cytokines and chemokines analyzed herein in a clinically relevant context. Alternative explanations for the elevated levels of HIV-inhibitory chemokines are needed to explain the putative beneficial effect of GBV-C. PMID- 15828090 TI - On shared services. PMID- 15828091 TI - The HSJ debate. Doctors should not share corporate responsibility. PMID- 15828092 TI - Buildings. Patient choice. Pick and mix. PMID- 15828093 TI - Buildings. Opinion: less is more. PMID- 15828094 TI - Buildings. Design awards. Brief encounter. PMID- 15828095 TI - Co-infection with HIV and hepatitis C: an overview. PMID- 15828096 TI - Current challenges in hepatitis C. PMID- 15828097 TI - The hepatitis C drug development pipeline. PMID- 15828098 TI - Clinical perspective: battling hepatitis C in our HIV-infected patients. PMID- 15828099 TI - My personal experience in being HCV/HIV co-infected & how I cured hepatitis C. PMID- 15828100 TI - What's pk (pharmacokinetics) got to do with it? Supplement introduction. PMID- 15828101 TI - The ABC's of pharmacokinetics. PMID- 15828102 TI - The nukes. Each one works a little differently. PMID- 15828103 TI - The non-nukes. Sustiva. Viramune. The two major players. PMID- 15828104 TI - The protease inhibitor drugs. PMID- 15828105 TI - Guys and girls. Interview with the founder of NYC's Straight Social Group. PMID- 15828106 TI - Online resources for HIV-positive heterosexuals. PMID- 15828107 TI - Small-town living and loving. How one woman infected at 50 went on to find happiness. PMID- 15828108 TI - New protease inhibitor TMC-114. Preliminary 24-week late breaker results of the phase II trial. PMID- 15828109 TI - The nebulous spirit. I started to read everything I could get my hands on about HIV and spirituality. PMID- 15828110 TI - Lowering the risk of HIV after sex or other exposure. PMID- 15828111 TI - Sex, lies and HIV: truth hurts. PMID- 15828112 TI - Looking to 2003. PMID- 15828113 TI - Dangerous liaison: club drug use and HIV/AIDS. PMID- 15828114 TI - HIV grudgingly yields some secrets (or, orthography and the retrovirus). 42nd Interscience Conference on Antimicrobial Agents and Chemotherapy, September 27 30, 2002. PMID- 15828115 TI - A single case of multidrug-resistant HIV and rapid disease progression. PMID- 15828116 TI - New guidelines for antiretroviral therapy in nonoccupational postexposure prophylaxis. PMID- 15828117 TI - Antiretroviral rounds. Resistance: how do you know if you don't know? PMID- 15828118 TI - Drug-induced hepatitis with saquinavir/ritonavir + rifampin. PMID- 15828119 TI - Immune reconstitution syndrome. AB - Immune reconstitution, or the reversal of HIV-related immune system decline, is one of the primary goals of highly active antiretroviral therapy (HAART). Reconstitution involves an increase in functional CD4 cells to guide the immune response against pathogens such as HIV, resulting in the suppression of viral load and other beneficial outcomes. However, immune reconstitution may trigger an inflammatory reaction in some people soon after they begin anti-HIV therapy and show signs of immunological improvement. Known as immune reconstitution syndrome (IRS) or immune reconstitution inflammatory syndrome (IRIS), this set of symptoms often resembles an AIDS-defining illness or other condition seen in people with HIV. While in most cases the symptoms of IRIS resolve after a few weeks, the syndrome may be severe or mistaken for true disease progression, and should be properly diagnosed and treated. PMID- 15828120 TI - Mortality trends: toward a new definition of AIDS? AB - HIV disease today looks very different than it did two decades ago when the first cases were described--at least in developed countries where highly active antiretroviral therapy (HAART) is widely available. Most HIV positive people receiving treatment no longer succumb to opportunistic illnesses (OIs) that take advantage of their ravaged immune systems. Instead, people with HIV now live longer and die from a wide variety of other causes. In other words, a growing number of positive people will die with HIV, but not of HIV. In the HAART era, HIV positive people and their health-care providers must now think about the cumulative impact of chronic HIV infection and the longterm side effects of treatment. In addition, as effective antiretroviral therapy extends the lives of people with HIV, they become prone to conditions that normally occur with greater frequency as people age (such as heart disease, diabetes, and osteoporosis) and progressive conditions that may take decades to cause significant illness or death (such as chronic viral hepatitis). Given the shifts in the types of conditions now seen in people with HIV, it may be time to rethink the definition of AIDS. PMID- 15828121 TI - Therapeutic vaccines: ready for prime (and boost) time? PMID- 15828122 TI - Women and HIV. Symptoms and quality of life in women. AB - Symptoms occur across the trajectory of HIV disease and can diminish the quality of life of women living with the virus. Untreated symptoms may also influence medication adherence and lead to complications of HIV and coexisting illnesses. Symptom management can address these concerns. A variety of self-care approaches should be incorporated into any plan of treatment, which ideally should be based on a partnership between the woman with HIV and her health-care providers. This article will address a range of issues concerning symptoms in women with HIV both in the U.S. and worldwide. PMID- 15828123 TI - Neurological complications of HIV/AIDS. AB - Neurological complications are common in HIV disease. The spectrum of neurological disorders is broad and involves the central nervous system, or CNS (brain and spinal cord) and the peripheral nervous system, or PNS (nerves outside the brain and spinal cord, and related muscle). Neurological disorders related to HIV often result in reduced quality of life and shortened survival, especially in people with more advanced HIV disease. Nevertheless, some neurological conditions are mild, readily treatable, or reversible. Several have become less common since the introduction of highly active antiretroviral therapy (HAART). And, despite the fact that many anti-HIV drugs are unable to cross the blood-brain barrier and penetrate the brain, recent data published in the Journal of Acquired Immune Deficiency Syndromes support the claim that HAART can improve some neurocognitive functioning. PMID- 15828124 TI - Headache and HIV. PMID- 15828125 TI - Analysis of the Par2 modifier of pulmonary adenoma formation in mice. AB - Inbred strains of mouse show various susceptibilities to spontaneous and chemical induced lung tumorigenesis. Genetic analyses have revealed that lung tumor susceptibilities of inbred mouse strains are governed by quantitative trait loci (QLTs) located on multiple chromosomes. A major lung tumor resistance QLT, designated pulmonary adenoma resistance 2 (Par2), was mapped to the mouse chromosome 18 independently by several groups and accounted for up to 60% phenotype variance between susceptible A/J and more resistant BALB/c strains. The authors recently conducted studies to positionally clone the Par2 gene. This review summarizes the effort and progress towards the identification of Par2 candidates. PMID- 15828126 TI - Souped-up yeast. PMID- 15828127 TI - ToxRAP makes the connection. PMID- 15828128 TI - Prenatal PAH exposure causes genetic changes in newborns. PMID- 15828129 TI - Choosing balance: congressional powers and the Partial-Birth Abortion Ban Act of 2003. PMID- 15828130 TI - Obesity exacerbates sepsis-induced inflammation and microvascular dysfunction in mouse brain. AB - OBJECTIVE: Obese patients with sepsis have higher morbidity and mortality than lean counterparts, but the mechanisms involved are unknown. The authors examined the inflammatory and thrombogenic responses of the cerebral microvasculature to sepsis induced by cecal ligation and perforation in obese and lean wild-type mice. METHODS: Leukocyte and platelet adhesion in cerebral microvasculature and behavioral responses were measured in wild-type and obese mice 4 h postperforation. P-selectin expression in different vascular beds was assessed 6 h postperforation. The effects of immunoblockade of P-selectin, ICAM-1, and CD18 on leukocyte and platelet recruitment were evaluated in obese septic animals. RESULTS: Cerebral venules of obese and wild-type mice assumed a proinflammatory and prothrombogenic phenotype 4 h post-perforation, with greatly exaggerated responses in obese mice compared to the lean counterparts. These enhanced responses were attenuated by blocking P-selectin, CD18, or ICAM-1. Obese mice also exhibited a more profound behavioral deficit after sepsis, which appears to be unrelated to the recruitment of leukocytes and platelets. Cecal ligation and perforation-induced P-selectin expression was greater in obese mice compared with lean counterparts. CONCLUSIONS: These findings suggest that the increased morbidity to sepsis in obesity may result from exaggerated microvascular inflammatory and thrombogenic responses that include the activation of endothelial cells with subsequent expression of adhesion molecules, such as P selectin. PMID- 15828131 TI - Source of gray residue in gas chromatograph injector liners, and how it can be prevented. PMID- 15828132 TI - Relationship between eluent composition and column operating pressure, especially as it relates to carrying out rapid reversed-phase separations. PMID- 15828133 TI - A new biodegradable poly-amino acid: alpha,beta-poly[(N-hydroxypropyl/aminoethyl) DL-aspartamide-co-L-lysine], a potential nonviral vector for gene delivery. AB - A new class of biodegradable poly-amino acid, alpha,beta-poly[(N hydroxypropyl/aminoethyl)-DL-Aspartamide-co-L-Lysine] (PHAAL), was synthesized by ring-opening of poly[succinimide-co-lysine](PSL) with n-propanolamine and ethylene diamine after thermal copolycondensation of DL-Aspartic acid and L lysine under reduced pressure. Different ratio feeds of PSL were obtained and characterized by 1H-NMR, Fourier transformed infrared spectroscopy, X-ray, thermogravimetric analysis and gel permeation chromatography experiments. As one of the polycationic materials, performed for gene delivery carrier, the PHAAL degradation experiment was carried out in PBS (10 mM, pH =7.4) and enzyme (papain, trypsine 1 mg/ml, 37 +/- 0.1 degree C) solution. PHAAL had lower cytotoxicity than polyethylenimine (25KDa) and poly-L-Lysine (30 KDa), in Hela, E.C.V.-304, Bcap 37 cell lines. Particle size and zeta, potential of PHAAL/DNA complexes were measured. Sizes ranged from 300-500 nm and zeta potentials were at -20 to 2,5 mV. The condensation ability of PHAAL for DNA was evaluated by agarose gel electrophoresis. The PHAAL could completely neutralize DNA at N/P ratio (w/w) 150:1. PMID- 15828134 TI - TRICARE; elimination of non-availability statement and referral authorization requirements and elimination of specialized treatment services program. Final rule. AB - This rule implements Section 735 of the National Defense Authorization Act for Fiscal Year 2002 (NDAA-02) (Pub. L. 107-107). It also implements Section 728 of the Floyd D. Spence National Defense Authorization Act for Fiscal Year 2001 (NDAA 01) (Pub. L. 106-398). Section 735 of NDAA-02 eliminates the requirement for TRICARE Standard beneficiaries who live within a 40-mile radius of a military medical treatment facility (MTF) to obtain a nonavailability statement (NAS) or preauthorization from an MTF before receiving inpatient care (other than mental health services) or maternity care from a civilian provider in order that TRICARE will cost-share for such services. Section 735 of NDAA-02, however, authorizes the Department of Defense to make exceptions to the elimination of the requirement for a NAS through the exercise of a waiver process under certain specified conditions. This section also eliminates the NAS requirement for specialized treatment services (STSs) for TRICARE Standard beneficiaries who live outside the 200-mile radius of a designated STS facility. This rule portrays the Department's decision to eliminate the STS program entirely. Finally, Section 728 of NDAA-01 requires that prior authorization before referral to a specialty care provider that is part of the contractor network be eliminated under any new TRICARE contract. PMID- 15828135 TI - Good actors in a bad movie. AB - Given the current swirl of activity surrounding charging, discounting, and collection activities by hospitals related to the uninsured, many healthcare executives feel like good actors in a bad movie. PMID- 15828136 TI - U.S. deaf education teacher preparation programs: a look at the present and a vision for the future. PMID- 15828137 TI - Educational programs for deaf students. Schools and programs in the United States. PMID- 15828138 TI - Educational programs for deaf students. Schools and programs in Canada. PMID- 15828139 TI - Educational programs for deaf students. Postsecondary programs. PMID- 15828140 TI - University and college programs for personnel in deafness. PMID- 15828142 TI - Advocacy, support, and rehabilitation programs. PMID- 15828141 TI - Programs for deaf-blind children and adults. PMID- 15828143 TI - Research on deafness. Doctoral dissertations. PMID- 15828144 TI - Imaging in medicine. PMID- 15828145 TI - Patients with grave maternal disorders. PMID- 15828146 TI - The conscience of the physician. PMID- 15828147 TI - Further reply to Ford. PMID- 15828148 TI - The case against premature induction. PMID- 15828149 TI - Genetic engineering and the moral status of non-human species. AB - Genetic modification leads to several important moral issues. Up until now they have been discussed from the viewpoint that only individual living beings, above all animals, are morally considerable. The standpoint that also collective entities such as species belong to the moral sphere have seldom been taken into account in a more thorough way, although it is advocated by several important environmental ethicists. The main purpose of this article is to analyze in more detail than often has been done what the practical consequences of this ethical position would be for the use of genetic engineering on animals and plants. The practical consequences of the holistic standpoint (focused on collective entities) of Holmes Rolson, III, is compared with the practical consequences of the individualistic standpoints (focused on individual living beings) of Bernard E. Rollin and Philipp Balzer, Klaus Peter Rippe, and Peter Schaber, respectively. The article also discusses whether the claim that species are morally considerable is tenable as a foundation for policy decisions on genetic engineering. PMID- 15828150 TI - Practical moral codes in the transgenic organism debate. AB - In one study funded by the United States Department of Agriculture, people from North Dakota were interviewed to discover which moral principles they use in evaluating the morality of transgenic organisms and their introduction into markets. It was found that although the moral codes the human subjects employed were very similar, their views on transgenics were vastly different. In this paper, the codes that were used by the respondents are developed, compared to that of the academically composed Belmont Report, and then modified to create the more practical Common Moral Code. At the end, it is shown that the Common Moral Code has inherent inconsistency flaws that might be resolvable, but would require extensive work on the definition of terms and principles. However, the effort is worthwhile, especially if it results in a common moral code that all those involved in the debate are willing to use in negotiating a resolution to their differences. PMID- 15828151 TI - Genomics: shifts in metaphorical landscape between 2000 and 2003. AB - This article examines the shifts and changes in the metaphors used to describe the human genome and the human genome project (HGP) between 2000 and 2003, with the year 2001 as a trigger for genomic and metaphorical reflection. We want to answer questions, such as: Did the findings announced in 2001 shake the metaphorical foundations on which the HGP had been built or not? Did novel metaphors capture the imagination of scientists and the public or did old metaphors survive throughout this period? What influence does the continuity or discontinuity in metaphorical framing of the HGP have on the public perception of the HGP as well as on its scientific understanding? To answer these questions we have systematically compared the metaphors used in one major scientific journal, Nature, and in one major UK newspaper, the online edition of the Guardian/The Observer during a period of two months around June 2000, February 2001 and April 2003. PMID- 15828152 TI - Genes, genomes and identity. Projections on matter. AB - This paper aims to show that references to genes and genomes are counterproductive in legal and political understandings of what it is to be human and a unique individual. To support this claim, I will give a brief overview of the many incompatible meanings the term 'identity' has gathered in reference to genes or genome in the contexts of biology and family ancestry, personal identity, species identity. One finds various and incompatible understandings of these expressions. While genetics is usually considered to deliver definitive knowledge about history and the future, genomics seems to work with more complicated relations between DNA, inheritance and phenotype. In genomics, 'identity' is no longer about identification and status markers but about individualization. Regulatory and legal documents project from traits to genomes, implying that individuality is at least represented, if not created, in a unique genome. Boundaries between humans and other animals, between different 'kinds' of humans, and between all individual humans are re-established via reference to the chemical matter of DNA. My analysis will show how this trend is a reactionary response to modern understandings of identities as social products and that it ignores new biomedical understandings of human bodies. PMID- 15828153 TI - Genomic patenting and the utility requirement. AB - This paper analyses the ways in which genomic knowledge is portrayed as useful knowledge in gene patenting in order to fulfil the 'utility'/'industrial applicability' requirement for patentability. It gives examples of utility claims in gene patents and asks whether genomics (as opposed to genetics) changes our ideas about what is useful and what can be patented. It puts forward a provisional classification of different types of utility and argues that merely identifying the physiological function of a gene diverges radically from our commonsense understanding of what it is for an invention to be useful. Furthermore, social, political and ethical issues inevitably arise when discussing the utility requirement, because an invention cannot be useful in isolation from a social context. PMID- 15828154 TI - Genomic susceptibility-testing and pregnancy: something old, something new. AB - This essay explores how testing for common and complex or genomic, as opposed to genetic, susceptibility to deep vein thrombosis both challenges and consolidates old social discourses on genes, gender and pregnancy. The nexus between genetics and reproduction usually crystallizes in the moral dilemma of selective termination. This essay examines online discussion among women with a genomic predisposition to deep vein thrombosis, which is associated with miscarriage and stillbirth. It explores the women's exchanges on what to 'do' in order to safely carry to term a foetus, which may always also have the genomic susceptibility. Interpreting DNA not in terms of predicting fate but of suggesting how to modify one's behaviour in order to give and care for life blunts its eugenic edge. However, this interpretation also shoulders discussants with the complicated and laborious responsibility of modifying themselves, their life styles and the life styles of their families--all of which falls within women's traditional labour of love in the privatized age of bioindividuality. PMID- 15828155 TI - Moral imagination in long-term care workers. AB - Our study focused on the cultural construction of dying and death in long-term care facilities. This article centers on direct care workers' perspective of residents' deaths. The data on which this article is based were gathered in a multi-year, multi-site study through formal ethnographic interviews, informal conversations, and on-site observations of residents and staff members. During fieldwork, we noticed an aptitude of direct care workers to deal with residents' deaths, which we named "moral imagination." The term is borrowed from other disciplines to describe a "way of seeing" residents. The case studies presented- that of three direct care workers: a dietary aide, a nurse aide, and an assistant activities director--are suggestive of workers in each category. Our study offers implications for future research concerning direct care workers' value to residents' quality of life. We also propose questions for long-term care facilities about standards of formal caregiving at the end of life. PMID- 15828156 TI - Sex selection and disability avoidance: is their opposed treatment conceptually consistent? AB - Sex selection and disability avoidance receive opposed treatment in bioethics literature, legislative practice and public opinion. However, some theorists question this state of affairs by drawing analogies between the harmful consequences of these practices. This paper shares their disapproval of gender selection and disability avoidance, but bases its resistance to these practices on an examination of the concepts of gender and disability. Here it identifies conceptual confusions as another cause of approval of sex selection and disability avoidance. Further, in clarifying the nature of the concepts at issue, and their relationship with the subjects that they apply to, this discussion highlights the existence of relevant analogies between the concepts of gender and disability. Here the social construction and universality of gender and disability allow their differential treatment to be resisted at the conceptual level, creating a strong foundation for more consequentialist arguments against sex selection and disability avoidance. PMID- 15828157 TI - Cochlear implants, the deaf culture, and ethics: a study of disability, informed surrogate consent, and ethnocide. AB - The use of cochlear implants in born-deaf infants addresses the issues of disability, proxy consent, and potential ethnocide of the Deaf culture. The ethical issues explored in this paper are: 1) the disability versus trait argument of deafness, 2) parents versus Deaf community in proxy consent, 3) justification for surgical intervention in a non-life threatening condition, and 4) justification for ethnocide. Decisions for non-competent individuals should be made to assure the child of an open future, with rights that need to be protected now, so that the child can exercise them later as an adult. Cochlear implants provide the potential of an open future and are morally justified on that basis. PMID- 15828158 TI - Human reproductive cloning: an analysis of the Andrews Report. AB - There is nothing like an overwhelming consensus of opinion to encourage a less than rigourous approach to analyzing complex ethical issues. Unfortunately, this is nowhere more apparent than in the discussion of human reproductive cloning contained in the federal House of Representatives Standing Committee on Legal and Constitutional Affairs' report on human cloning, released last August. The report may well fulfil the first half of its project, namely the empirical task of adequately summarizing and categorizing the various submissions made to the Committee. However, it is clearly inadequate as a discussion of the ethical and legal permissibility of human reproductive cloning. PMID- 15828159 TI - Editorial. PMID- 15828160 TI - Genetic manipulation and our duty to posterity. AB - To what extent should scientists, doctors and the community be constrained in their decision-making by a duty to posterity? How should we as a community balance our desire to benefit the present generation against the need not to irretrievably harm our successors? These questions are discussed with particular reference to genetic research and treatment that may have great potential for people suffering from genetic disease but may cause inherited changes in future generations, either deliberately or inadvertently. We conclude that the community should take account of the interests of its successors but this should not immobilise us in the decisions we make for the present. PMID- 15828161 TI - Euthanasia, double effect, and proportionality. AB - I discuss the Principle of Double Effect (PDE) as a means of exploring aspects of the motivation of active voluntary euthanasia (a.v.e). It is argued that the objective of a.v.e. is not death but the relief of suffering. Nor is death the means, it is a concomitant. I entertain no hope of convincing the typical proponent of PDE that a.v.e. is morally acceptable. However, I point out that a central strength of the PDE is its insistence on due proportionality between outcomes. When a.v.e. is appropriate, as sometimes it is, death is not out of proportion to the relief of suffering. PMID- 15828162 TI - Patient-centred equipoise and the ethics of randomised controlled trials. AB - The ethical pre-condition of randomised controlled trials is, at present, the presence of equipoise. This refers to an opinion of the investigator that there is uncertainty as to the merits of the treatments being compared. It is argued that since the decision to enrol is the potential subject's, the investigator's opinion is not ethically relevant. It is proposed instead that equipoise be patient-centred, and that a trial is in equipoise for a patient when enrolling gives them the same chance of a good outcome as not enrolling. It is shown that trial enrolment is normally easily the best strategy for achieving a good outcome, and that trials currently thought to be unethical may be in patient centred equipoise. This is so for three reasons: (1) patients in trials receive superior care, (2) trial enrolment minimises the risk of being a victim of a therapeutic disaster and (3) health professionals make mistakes, and a 50% chance of receiving the worse treatment until a trial reports is always better than any chance of receiving the worse treatment indefinitely. It is concluded that so long as the standard of professional conduct is the furtherance of patients' objective interests randomised controlled trials are ethical. PMID- 15828163 TI - Inhibition of growth of Mycobacterium smegmatis and of cell wall synthesis by D serine. AB - d-Serine inhibited the growth of Mycobacterium smegmatis and induced the morphological alteration of the bacilli. The growth inhibitory action of d-serine was partially reduced by an equimolecular concentration of d-alanine. The combination of glycine with d-alanine reversed the growth inhibition produced by d-serine more than did d-alanine alone. In cells cultured in the presence of d serine, the amounts of alanine, diaminopimelic acid, and glycine inserted into the cell wall mucopeptide were reduced, and serine was increased. The intracellular accumulation of a precursor of cell wall mucopeptide was increased by d-serine, and this accumulation was reduced by d-alanine. d-Serine competed with glycine for incorporation into the cell wall mucopeptide. The incorporation of l-aspartic acid into diaminopimelic acid residues in the cell wall mucopeptide was markedly inhibited by d-serine. Three mutants resistant to d-serine were isolated by nitrosoguanidine treatment. In these mutants the effects of d-serine on the sites of cell wall mucopeptide synthesis were all reduced. Thus, d-serine inhibition of the growth is due to replacement of glycine residues of the cell wall mucopeptide with d-serine and inhibition of the cell wall synthesis by blocking the formation of d-alanine and diaminopimelic acid. PMID- 15828164 TI - Transamidinase activity of hyphal fractions of a streptomycin-producing Streptomyces griseus strain and enhancement of enzyme activity in a nonproducing mutant. AB - The presence of transamidinase (TA-se) was demonstrated in Streptomyces griseus strain 45 which does not produce streptomycin (SM). This enzyme is involved in the biosynthesis of streptidine, a component of the SM molecule. TA-se is generally thought to be present only in SM-producing Streptomyces strains. The recent results are, however, in agreement with out previous data which showed the presence of streptidine in the isolated cell wall of SM-nonproducing strain 45. The TA-se activity of this strain is very low, but we were able to increase it by addition of arginine or by increasing the incubation with myo-inositol. The addition of these substances to the cultures of SM-nonproducing S. griseus strains, which hitherto were deficient in TA-se, may serve as a method for detecting the presence of this enzyme. In an incubation system in which the hyphae of an SM-producing strain of S. griseus had been blended and separated by density gradient centrifugation, certain hyphal fractions had been combined, and their SM production and TA-se activity had been measured, we were unable to find any close correlation between TA-se activity and SM-producing capacity. The hyphal fractions and their combination did not show strict parallelism between TA se activity and the level of SM production. PMID- 15828165 TI - In vitro antimalarial activity of nucleic acid precursor analogues in the simian malaria Plasmodium knowlesi. AB - Analogues of nucleic acid precursors were screened for antimalarial activity in Plasmodium knowlesi by using an in vitro culture system. Activity was assessed by the degree of inhibition of incorporation of l-[methyl-(14)C]methionine into protein and of [8-(14)C]adenosine and [6-(14)C]orotic acid into ribonucleic acid and deoxyribonucleic acid. The incorporation of adenosine or orotic acid was effectively inhibited by many of the compounds, including 3' analogues of purine nucleosides, many of the 6-position analogues of purine bases and nucleosides, and 5-position analogues of orotic acid. Only a few compounds inhibited methionine incorporation into protein, and in each instance adenosine or orotic acid incorporation also was inhibited. Some compounds inhibited adenosine or orotic acid incorporation into both ribonucleic acid and deoxyribonucleic, whereas other analogues inhibited incorporation into one nucleic acid only. The qualitative and quantitative differences suggest that this experimental system may be appropriate for investigation of metabolic pathways of the malaria parasite, as well as for demonstration of antimalarial activity of candidate antimalaria drugs. PMID- 15828166 TI - Multiple antibiotic resistance in Neisseria gonorrhoeae. AB - The sensitivities of gonococcal isolates to six antibiotics were determined for gonococci isolated in Philadelphia in 1972. The degree of association between susceptibilities to any two antibiotics was determined (coefficient of correlation). The correlation between penicillin and tetracycline (r = 0.75) was almost as good as that between two penicillins, penicillin G and ampicillin (r = 0.85), but the difference was statistically significant. The lowest correlation found was between erythromycin and chloramphenicol (r = 0.62), two antibiotics seldom used in gonorrhea therapy. In addition, gonococci most resistant to one antibiotic were the most likely to be multiply resistant. This was found with respect to penicillin, tetracycline, erythromycin, and chloramphenicol. Approximately 40% of gonococci classified as "most resistant" (exceeding the resistance of 75% of all isolates) to one antibiotic were also "most resistant" to three others. Finally, multiply resistant mutants were isolated by selection for resistance to either penicillin or tetracycline. These results provide evidence for the existence of a common mechanism for multiple antibiotic resistance in the gonococcus. PMID- 15828167 TI - Rapid radiometric method of testing susceptibility of mycobacteria and slow growing fungi to antimicrobial agents. AB - [(3)H]uracil incorporation into the ribonucleic acid (RNA) of mycobacteria and [(3)H]guanine incorporation into the RNA of fungi were used as measures of cell viability in the presence of several antimicrobial agents. There appeared to be an excellent correlation between traditional susceptibility tests, which required 1 to 4 weeks, and tests of the inhibition of incorporation of RNA precursors, which were completed in 4 to 48 h. The radiometric method may be useful in rapidly determining the susceptibility of slow-growing organisms to antimicrobial agents. PMID- 15828168 TI - Variables influencing the in vitro susceptibilities of herpes simplex viruses to antiviral drugs. AB - Some of the variables in a microtiter tissue culture method for estimating antiviral susceptibility of herpes simplex viruses (HSV) were studied. Six antiviral drugs (5-iodo-2'-deoxyuridine, cytosine arabinoside, 9-beta arabinofuranosyladenine, isoprinosine, virazole, and 2-deoxy-d-glucose) and three tissue culture cell lines (primary rabbit kidney, WI-38, and rat brain) were studied. Four different inocula (10, 100, 1,000, and 2,000 mean tissue culture doses) and nine isolates of HSV were tested. Minimal inhibitory concentration was defined as a complete inhibition of cytopathogenic effect in tissue culture cells after 72 h of incubation. There was a direct relationship between the viral inocula and the minimal inhibitory concentration. There were also differences between the three tissue culture cell lines and among the different strains of herpesvirus. This was not type specific, and the strain differences were not consistent in different tissue culture lines. Cytosine arabinoside appeared to be the most active drug in vitro, with 5-iodo-2'-deoxyuridine intermediate in activity between cytosine arabinoside and 9-beta-arabinofuranosyladenine; no significant cytotoxicity was encountered with these drugs. Isoprinosine, virazole, and 2-deoxy-d-glucose were not effective against any of the nine strains of HSV tested in this in vitro system; a comparison with previous studies indicates that their activity may be cell dependent. These results emphasize the importance of careful evaluation of in vitro test methods for the susceptibility testing of antiviral agents against HSV. Marked variability between strains and in vitro test systems caution against the application of these methods to in vivo situations until further studies are carried out. PMID- 15828169 TI - Inhibition of [14C]chloramphenicol binding to Escherichia coli ribosomes by erythromycin derivatives. AB - The effect of erythromycin A and 35 analogues of erythromycin A on [(14)C]chloramphenicol binding to Escherichia coli ribosomes was evaluated. Substitutions on various portions of the erythromycin molecule were made with retention of ability to bind to ribosomes. Specifically, substantial activity in interference with [(14)C]chloramphenicol binding was retained upon removal of the cladinose and various substitutions on the 3-hydroxyl, the oxime, and 2-hydroxyl groups. Erythromycin analogues with relatively poor binding activity to ribosomes could be detected. This assay can be used alone or in conjunction with microbiological assays for screening of active analogues. It permits an estimate of the general binding activity of compounds rapidly and directly. The assay reflects the ability of the compounds to interact with their target organelle, the ribosome, and may serve as a useful adjunct in developing new compounds. PMID- 15828170 TI - Pleiotropic antibiotic resistance mutations associated with ribosomes and ribosomal subunits in Mycobacterium smegmatis. AB - Viomycin-resistant strains isolated from Mycobacterium smegmatis demonstrated pleiotropic resistance to tuberactinomycin-N, capreomycin, streptomycin, and kanamycin as a result of mutational alteration of ribosomes, even though they were selected for resistance to a single antibiotic. The pleiotropic drug resistance of three mutants isolated by stepwise selection for resistance to viomycin was due to alteration of the 30S ribosomal subunit. One mutant, strain A, isolated independently by multiple-step selection to viomycin resistance, was resistant to viomycin, tuberactinomycin-N, and capreomycin through an alteration of the 50S ribosomal subunit, whereas it was sensitive to kanamycin but resistant to streptomycin through an alteration of the 30S ribosomal subunit. Three streptomycin-resistant strains, which were isolated by one-step selection at a high concentration of streptomycin, did not show significant co-resistance to any other antibiotics tested in culture and cell-free systems; streptomycin resistance in these mutants was localized on the 30S ribosomal subunit. PMID- 15828171 TI - Rapid microassays for clindamycin and gentamicin when present together and the effect of pH and of each on the antibacterial activity of the other. AB - Gentamicin was measured in the presence of clindamycin by using as the assay organism a strain of Staphylococcus aureus that was resistant to clindamycin; clindamycin was measured in the presence of gentamicin by using Clostridium perfringens as the assay organism. Both assays were in agar diffusion systems, were rapid (2 to 4 h), and had errors of less than 10%. Neither antibiotic antagonized or synergized with the other against a variety of organisms. The pK of clindamycin was 7.3, and the pK of gentamicin was 8.2. Gentamicin was 100 or more times as active in alkaline medium against both gram-positive and gram negative organisms, but clindamycin showed a striking pH effect on activity only with gram-positive organisms. PMID- 15828172 TI - R-mediated beta-lactamases and episomal resistance to the beta-lactam drugs in different bacterial hosts. AB - Ten penicillinase plasmids of varying taxonomic origin were studied after transfer to a variety of bacterial hosts. Nine of the ten plasmids specified enzymes with the following identical, or very similar, properties: substrate profile, molecular weight, susceptibility to heat and inhibitors, and electrophoretic mobility, i.e., TEM-like enzymes. The tenth R-mediated beta lactamase was a cephalosporinase. Plasmids with TEM-like enzymes mediated resistance patterns identical towards the beta-lactam drugs, whereas the resistance pattern of the cephalosporinase plasmid was distinctly different. Expression of enzyme and resistance had a dual R-factor and host specificity. Escherichia coli K-12 and Salmonella typhi constituted one group of the same R factor phenotype expressions. Most, but not all, penicillinase plasmids exhibited in Proteus PM1 a considerably lower order of beta-lactamase activity and an even lower order of resistance to the beta-lactam drugs than the previous two hosts. This difference was most pronounced for the resistance to carbenicillin, which was mediated by the plasmids specifying the synthesis of TEM-like enzymes. Release by osmotic shock was complete in the host E. coli K-12 for the TEM-like enzymes, but was lower for the cephalosporinase and minimal or negative in the PM1 host. Crypticity factor for benzylpenicillin, ampicillin, and carbenicillin was not related to the increase in resistance mediated by the penicillinase plasmids in both K-12 and PM1 hosts. Inoculum size effects for the penicillins and 6-aminopenicillanic acid were higher in PM1 than in K-12 R(+) cultures. The expression of penicillinase plasmids in wild-type bacteria was strain specific and not species specific. For two plasmids of different phenotypes for beta lactamase activity (and resistance) in K-12 and PM1 hosts, a positive correlation was found between their phenotype and the relative amount of episomal deoxyribonucleic acid, as detected by ethidium bromide density gradient centrifugation. This is interpreted as indicating differences in the mode of replication of the plasmids in the two hosts. PMID- 15828173 TI - Relation of cell wall lipid content of Serratia marcescens to resistance to antimicrobial agents. AB - Serratia marcescens strains were divided into three groups on the basis of antimicrobial sensitivity and pigment production. Group I, nonpigmented, was resistant to most antibiotics. Group II, nonpigmented, was susceptible to many antimicrobial agents, as was group III which was pigmented. Representative organisms of each group were examined for all lipid content. There were no significant differences in total lipid, phospholipid, or fatty acid esters among the three groups. Differences in susceptibility to antibiotics in Serratia do not seem to be explained on the basis of wall lipid content. PMID- 15828174 TI - Purification and characterization of syringacin 4-A, a bacteriocin from pseudomonas syringae 4-A. AB - Syringacin 4-A, a bacteriocin produced by Pseudomonas syrinagae 4-A, was obtained by induction with ultraviolet irradiation or mitomycin C. Approximately 1,000 fold purification of the bacteriocin was achieved by manganous chloride precipitation, differential centrifugation, and chromatography on hydroxyapatite columns. The purified syngacin was homogeneous on hydroxyapatite columns and sucrose density gradients; it also sedimented as a single entity in the analytical ultracentrifuge. The buoyant density of purified syringacin in cesium chloride was 1.294 g/ml. The sedimentation coefficient was calculated as 120S, and the diffusion coefficient was 6.49 x 10(-8) cm(2)/s. The molecular weight was calculated as 1.6 x 10(7) from physical data and 1.7 x 10(7) from biological data. The syringacin was composed of about 88.4% protein, 8.5% arabinose, 2.2% galacturonic acid, and 0.7% glucosamine. Amino acid analysis indicated a predominance of leucine (12.1%), aspartic acid (12.2%), and glutamic acid (12.7%). The ultraviolet spectrum showed a maximum absorbance peak at 276 nm. The syringacin was heat and alcohol sensitive, but resistant to trypsin, chymotrypsin, carboxypeptidase, Pronase, protease, lysozyme, steapsin, deoxyribonuclease, and ribonuclease. Maximum pH stability was between 5 and 8. Crude bacteriocin was stable at room temperature for at least a year, and purified material was stable for at least 3 months at 4 C. PMID- 15828175 TI - Combined activity of sulfamethoxazole, trimethoprim, and polymyxin B against gram negative bacilli. AB - The activity of the three two-drug combinations of sulfamethoxazole (SMX), trimethoprim (TMP), and polymyxin B (PB) against 52 clinical isolates of gram negative bacilli was studied by a "checkerboard" agar dilution method. The organisms studied included strains of Enterobacter spp., Klebsiella pneumoniae, Serratia marcescens, Providence, Proteus, and Pseudomonas aeruginosa. The majority of these isolates were resistant to at least two of the three agents used in the combined studies and to the most commonly used antimicrobials. The TMP-PB combination demonstrated enhanced activity more frequently than the other two-drug combinations, showing synergism or addition in 85% of the combined studies; indifference or antagonism was also observed least frequently with TMP PB. The great majority (83%) of Enterobacter-Klebsiella-Serratia isolates were susceptible to enhanced activity of all combinations. Proteus-Providence isolates were frequently susceptible (63%), but combined activity was indifferent or antagonistic against 60% of P. aeruginosa. Twelve isolates were selected for "killing-curve" assays in which an inoculum was incubated with SMX, TMP, and PB individually and in various two- and three-drug combinations. Surviving bacteria were counted at timed intervals over 24 h of incubation. The triple combination (SMX-TMP-PB) was synergistic against 9 of 12 isolates, whereas TMP-PB and SMX-PB showed synergism against 5 and 3 isolates, respectively. These data suggest that, although TMP-PB will often show enhanced activity against the gram-negative bacilli studied here, optimal antibacterial activity will be demonstrated when the three-drug combination is used. PMID- 15828176 TI - Lack of activity of sulfamethoxazole and trimethoprim against anaerobic bacteria. AB - The activity of sulfamethoxazole (SMX), trimethoprim (TMP), and the combination of the two was determined against a variety of anaerobic bacteria. Brucella agar was somewhat inhibitory for SMX and TMP but activity was good and equivalent in Diagnostic Sensitivity Test Agar (Oxoid) and Mueller-Hinton agar and the latter was selected for use in these studies. Agar dilution susceptibility tests showed that 95 of 98 anaerobic isolates were resistant to >/=100 mug of SMX per ml and 85 were resistant to >/=6.25 mug of TMP per ml. "Checkerboard" agar dilution studies of combined activity showed that 66 of 72 isolates were resistant to >/= (100 mug of SMX per ml + 6.25 mug of TMP per ml) and only six isolates were susceptible to the synergistic activity of the combination. The majority of 32 isolates tested by the disk diffusion method were also resistant to SMX and TMP individually and to the combination 25-mug disk. Correlation between agar dilution minimal inhibitory concentration and disk zone size results was in general good for individual agents. Four Bacteroides fragilis isolates were inhibited by the combination 25-mug disk but were resistant to SMX + TMP by agar dilution "checkerboard." This discrepancy may have been due to different incubation periods since disk results also showed resistance when read after 48 h (as is done with agar dilution) rather than the standard 24 h for disk tests. These studies suggest that SMX and TMP, either individually or in combination, are not active against the great majority of anaerobic bacteria. PMID- 15828177 TI - Determination of plaque inhibitory activity of adenine arabinoside (9-beta-D arabinofuranosyladenine) for herpesviruses using an adenosine deaminase inhibitor. AB - The in vitro susceptibility of type 1 and type 2 strains of Herpesvirus hominis to 9-beta-d-arabinofuranosyladenine (adenine arabinoside, ara-A) was measured in a system where deamination was inhibited. Under these conditions, it was possible to measure the activity of low concentrations of ara-A. It was determined that plaque inhibitory concentration for type 1 viruses was less than 3 mug/ml for all strains tested. The plaque inhibitory concentration for 7 of 10 type 2 strains was also less than 3 mug/ml. The method used identified and controlled the interaction between antiviral agent (ara-A) and the indicator system, human skin fibroblastic cells. Otherwise, metabolism of ara-A resulted in rapid enzymatic degradation and loss of antiviral activity. PMID- 15828178 TI - Polyene-resistant mutants of Aspergillus fennelliae: sterol content and genetics. AB - Mutants of Aspergillus fennelliae (Neosartorya fennelliae) resistant to relatively high levels of amphotericin B and low levels of nystatin were obtained by successive transfers of wild type in the presence of increasing concentrations of the polyenes. The resistance of the mutants to the polyenes was accompanied by both qualitative and quantitative changes in the sterol composition of the cells. Those resistant to amphotericin B (AF5-AB1 and p-AB1) lacked ergosterol, the major sterol of the wild type, but contained a new sterol clearly distinguished by the pattern of ultraviolet spectrophotometry and thin-layer chromatography. The mutants resistant to nystatin, however, contained both ergosterol and a new sterol, but the former was produced in a much reduced amount, as compared with the wild type. Genetic analysis indicated that the lack of ergosterol is closely associated with a reduced growth rate, poor asexual reproduction, and the loss of sexual reproduction. Growth studies revealed that the addition of ergosterol to the media did not affect the growth pattern of the mutants. Mutants resistant to amphotericin B showed an increased minimal inhibitory concentration for nystatin, pimaricin, and filipin. Mutants resistant to nystatin, however, conferred increased minimal inhibitory concentration for pimaricin and filipin but not for amphotericin B. PMID- 15828179 TI - In vitro susceptibility of Haemophilus influenzae to eight antibiotics. AB - Freshly isolated strains of Haemophilus influenzae type B were studied for their in vitro susceptibility to eight antibiotics with a low concentration inoculum. No organisms were identified as being resistant to ampicillin, but minimal inhibiting concentrations for that antibiotic were somewhat higher than reported previously for a similar method. Carbenicillin and ticarcillin resembled ampicillin in activity, and the three agents were more effective on a weight basis than the other agents tested. PMID- 15828180 TI - Antigen specific for bacteria resistant to tetracycline. AB - Antisera produced against envelope fractions of tetracycline-resistant Escherichia coli and supernatant fluid of disrupted cells of tetracycline resistant Salmonella typhi were adsorbed with the corresponding fractions of susceptible cultures. With these sera, cross-reacting antigens were demonstrated in the following fractions obtained from tetracycline-resistant strains: envelopes of E. coli, supernatant fluid of S. typhi, and envelopes and supernatant fluid of Pseudomonas aeruginosa. These antigens were not demonstrable in the corresponding fractions from tetracycline-susceptible strains. PMID- 15828181 TI - Distribution in nature of R factors that increase susceptibility to rifampin of rif-r mutants in Escherichia coli. AB - Among 117 gram-negative bacteria isolated from pathological materials, 46 were found to carry antibiotic resistances transferable to Escherichia coli K-12; we therefore concluded that they carry infectious R factors. When transferred to a type of rifampin-resistant mutant of E. coli, all these R factors decreased the resistance to rifampin, but only 10% of them lowered the resistance to one-tenth or less that of the isogenic R(-) strain. The relevance of these facts for the epidemiology of R factors in gram-negative bacteria is discussed. PMID- 15828182 TI - Butirosin compared with gentamicin in vitro and in vivo. AB - Butirosin (BTN) (P. W. K. Woo, G. L. Coffey, H. W. Dion, S. A. Fusari, and G. D. Senos, U. S. Patent 3,541,078, 1970) is a new aminoglycoside antibiotic notably active against opportunist bacterial species within Pseudomonas, Klebsiella, Enterobacter, Serratia, and Proteus. Numerous comparative tests were carried out with BTN and gentamicin (GTM) in vitro and in experimental infections in mice. BTN was more active in Mueller-Hinton broth than in agar, but its activity was lessened at acid pH or under anaerobiosis, as has been observed with other aminoglycosides. In standard agar diffusion tests, inhibition zones greater than 12 mm around 30-mug BTN disks generally denoted susceptibility, equivalent to minimal inhibitory concentrations [Formula: see text] 25 mug/ml. Cross-resistance between BTN and GTM occurred in a variable manner, with a number of bacterial strains strongly resistant to GTM being moderately susceptible to BTN. In mice, after a single subcutaneous injection, absorption of both antibiotics was rapid, with peak serum levels occurring in 15 min; this was followed by rapid elimination with estimated serum half-lives of about 20 min for each. After peroral administration of high doses in mice, there was no appreciable absorption of BTN. Several tests were carried out to compare BTN and GTM with respect to minimal inhibitory concentrations in vitro, acute subcutaneous median mouse protective doses, peak serum levels at such doses, and the therapeutic ratios derived from acute median protective and lethal doses. Although GTM usually proved to be more potent antibacterially on a weight basis, observations on BTN indicated a superior effectiveness in terms of therapeutic ratios. PMID- 15828183 TI - Macromolecule metabolism and photosynthetic functions in blue-green algae treated with virginiamycin, an inhibitor of protein synthesis. AB - The M component of virginiamycin inhibited growth of Plectonema boryanum under both photoautotrophic and heterotrophic conditions. Though the S component of this antibiotic had no apparent activity per se, it enhanced the inhibitory action of its partner. Cells incubated with suitable concentrations of either M or M + S stopped growing and lysed. Loss of the colony-forming capacity occurred quickly in the presence of M + S and slowly in the presence of M alone. Virginiamycin M inhibited protein synthesis in autotrophically and heterotrophically growing Plectonema. This effect was very rapid and could be reversed by removing the antibiotic. The S component did not block the incorporation of amino acids into proteins, but prevented the reversibility of the inhibitory effect of M. Virginiamycin M or S did not affect the photosynthetic oxygen development (Hill's reaction) in Plectonema. Moreover, carbon dioxide photoassimilation and formation of chlorophyll were inhibited only after an appreciable lag. Deoxyribonucleic acid synthesis was blocked virtually without delay by virginiamycin M. Since virginiamycin inhibited protein synthesis in a similar fashion in the unicellular Anacystis nidulans, as well as in the filamentous P. boryanum, the mechanism of action of this antibiotic is probably the same in all blue-green algae. PMID- 15828184 TI - Antibiotic G-418, a new Micromonospora-produced aminoglycoside with activity against protozoa and helminths: fermentation, isolation, and preliminary characterization. AB - Antibiotic G-418 is a new aminoglycoside produced as the major component by a new species of Micromonospora, M. rhodorangea NRRL 5326. The antibiotic is prepared by submerged fermentation in a soybean-dextrin medium. Antibiotic G-418 is adsorbed on a cationic-exchange resin and separated from other impurities by passing it down a Dowex (1 x 2) resin column. The antibiotic, which contains 2 deoxystreptamine, has broad-spectrum antibacterial activity and is highly active against protozoa, amoebae, tapeworm, and pinworm infections in mice. This report describes the taxonomy of the organism, and fermentation, isolation, and preliminary characterization of antibiotic G-418. PMID- 15828185 TI - Comparative serum levels and protective activity of parenterally administered cephalosporins in experimental animals. AB - Six cephalosporin antibiotics were administered subcutaneously to mice at a level of 20 mg/kg. The serum levels of each were determined at five time intervals ranging from 5 to 120 min after dosing. Urinary recovery and the presence of active metabolites in mouse urine were determined. The peak serum levels and serum half-lives in mice were found to be positively correlated with the mean effective dose values obtained after lethal challenge with Escherichia coli. The administration of cefazolin and cephanone resulted in the highest serum level and the best protection. Good protection was obtained with cephaloridine despite somewhat lower serum levels. The cephalosporins with the acetoxy side chain (cephalothin, cephapirin, and cephacetrile) showed lower serum levels and the poorest protection. Cefazolin, cephaloridine, and cephalothin serum levels were also determined in dogs, squirrel monkeys, and rabbits. A mixed response was obtained in these species, with cefazolin peak serum levels being highest in rabbits and cephaloridine peak highest in dogs. PMID- 15828186 TI - Mutanolysin, bacteriolytic agent for cariogenic Streptococci: partial purification and properties. AB - Mutanolysin partially purified from the culture filtrate of Streptomyces globisporus 1829 consists of two main lytic enzymes with an isoelectric point near pH 8.5 and 10, respectively, and proteolytic enzyme is associated with the latter lytic enzyme. Mutanolysin exhibited maximal lytic activity at 60 C in the pH range 6.5 to 7.0 and was stable at 50 C in the acid range. N-bromosuccinimide caused complete inhibition of lytic activity at 1 mM, whereas calcium and magnesium ions at the same concentration caused activation. Mutanolysin had lytic or bactericidal activity against the living cells of Streptococcus mutans, Streptococcus salivarius, Streptococcus sanguis, Lactobacillus acidophilus, and Actinomyces viscosus, which are considered to be etiologic agents of dental caries, but had no activity against S. aureus and all gram-negative strains tested. The lytic activity was well retained in human saliva. Digestion of the cell walls of S. mutans BHT by mutanolysin was accompanied by the liberation of free amino groups and reducing sugars. Mutanolysin may be expected to be a useful agent for dental caries control. PMID- 15828187 TI - Diphtheria carriers and the effect of erythromycin therapy. AB - One hundred and fourteen Corynebacterium diphtheriae, toxigenic, gravis type, pharyngeal carriers were identified during a diphtheria epidemic in Elgin, Texas. All carriers were treated with erythromycin estolate, 1 g/day in divided doses for 6 days. Serial pharyngeal cultures were obtained in order to monitor the bacteriological response. Seventy-two carriers had positive cultures immediately prior to the start of therapy, and only these individuals were considered in the analysis of the effects of erythromycin. Forty-eight hours after institution of therapy, 96% of the carriers had become culture negative; all were negative by the 4th day of therapy, and all remained culture negative while taking the drug. Two days after cessation of therapy, all but one (99%) were culture negative. However, upon reculture 2 weeks later, 15 (21%) had relapsed to the carrier state. There were no significant differences in the serum diphtheria antitoxin levels, immunization status, age, sex, or socioeconomic status of those who relapsed and those who remained culture negative. This study demonstrates that erythromycin is effective in converting carriers to culture-negative status, but when given for only 6 days it is associated with large numbers of relapses. Because previous studies have not included follow-up cultures 2 weeks after therapy, it is suggested that all C. diphtheriae carriers be treated with either erythromycin or penicillin and that all be recultured at a minimum of 2 weeks after completion of therapy to assure eradication of the diphtheria organisms. PMID- 15828188 TI - Cefoxitin, a semisynthetic cephamycin antibiotic: antibacterial spectrum and resistance to hydrolysis by gram-negative beta-lactamases. AB - The in vitro activity of cefoxitin, 3-carbamolyloxymethyl-7-alpha-methoxy-7[2-(2 thienyl)acetamido]-3-cephem-4-carboyxlic acid, was investigated. Activity against gram-positive organisms was less than that of cephalothin and cephloridine. It was highly active against gram-negative bacilli, with activity against Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae equal to that of currently available cephalosporins. In addition, it was active against certain Enterobacter strains, Serratia marcescens, indole-positive Proteae and Herellea. The strains of these latter bacteria were strains susceptible to carbenicillin and ticarcillin. Pseudomonas aeruginosa and other Pseudomonas species were resistant. Changes in pH, inoculum size, and type of growth medium had no significant effect on the activity of the antibiotic. Cefoxitin was highly resistant to hydrolysis by various types of gram-negative beta-lactamases. The precise role of resistance to beta-lactamase hydrolysis varied from strain to strain. Bacterial resistance to cefoxitin was not necessarily related to hydrolysis of the antibiotic. However, the resistance of cefoxitin to hydrolysis did contribute to its activity. Cefoxitin could function as an inducer of beta lactamase activity and effectively bound to purified beta-lactamases. PMID- 15828189 TI - Cefamandole, a cephalosporin antibiotic with an unusually wide spectrum of activity. AB - The in vitro activity of cefamandole, 7-d-mandelamido-3-(1 methyl-1H-tetrazol-5 yethiomethyl)-3-cephem-4 carboxylic acid, was investigated. The majority of streptococci and pneumococci were inhibited by 0.1 mug/ml. Activity against Staphylococcus aureus was below that of cephalothin, but most strains were inhibited by 0.4 mug/ml. Enterococci were not inhibited by less than 25 mug/ml. Cefamandole was very active against most members of the Enterobacteriaceae, with 70% of Escherichia coli, 86% of Klebsiella pneumoniae, and 88% of Proteus mirabilis inhibited by 1.6 mug/ml. A number of Enterobacter aerogenes, Proteus vulgaris, and Serratia marcescens strains were inhibited by less than 25 mug/ml. Pseudomonas species were resistant. Cefamandole was more active than cephalothin, cephaloridine, or cephalexin against members of the Enterobacteriaceae. Inoculum size and type of medium had variable effects on the minimal inhibitory concentration, depending upon the particular strain tested. Strains of Enterobacter, Serratia, indole-positive Proteus species showed the greatest inoculum effect. Similarly, these species showed a several-fold difference of bactericidal and inhibitory levels. Resistance of cefamandole to hydrolysis by gram-negative beta-lactamase played a partial role in its activity, but some strains that hydrolyzed the agent were susceptible. PMID- 15828190 TI - Characteristics and expression of tetracycline resistance in gram-negative bacteria carrying the Pseudomonas R factor RP1. AB - The Pseudomonas R factor RP1 determines an inducible tetracycline resistance similar to that described for Escherichia coli R factors. The level of RP1 determined resistance measured by minimal inhibitory concentration testing is dependent on the host bacteria and corresponds to the magnitude of decrease in tetracycline accumulation by RP1-containing organisms. The tetracycline resistance mechanism is inactivated at low temperatures. The effect of metabolic inhibitors on tetracycline accumulation by susceptible organisms under various conditions indicates a possible nonspecific effect of cellular energy metabolism on tetracycline uptake. PMID- 15828191 TI - Gentamicin resistance in Pseudomonas aeruginosa: R-factor-mediated resistance. AB - By disk diffusion antimicrobial susceptibility testing, 11% of 313 consecutive strains of Pseudomonas aeruginosa, examined during July to October 1973, were resistant to gentamicin (minimal inhibitory concentration 12.5 to >100 mug/ml), and a further 31% were moderately resistant (6.25 to 12.5 mug/ml) to gentamicin at the University of Alberta Hospital in Edmonton, Canada. Of 45 gentamicin resistant strains from that hospital, none possessed R-factors or gentamicin inactivating enzymes. Eight of 13 strains obtained from three American sources, which contained gentamicin-acetylating (12 strains) or -adenylating (1 strain) activity, conjugally transferred both gentamicin resistance and antibiotic inactivating activity. P. aeruginosa recipients were much more effective for detection of transferable gentamicin resistance than Escherichia coli recipients, although not all P. aeruginosa were equally as effective as recipients. One strain, POW 151, transferred resistance to both carbenicillin and gentamicin as well as to several other antibiotics. R-factors detected belonged to P-2 and P-3 (Com 6, C) incompatibility groups. Expression of gentamicin resistance due to acetylation of gentamicin was subject to marked phenotypic lag, especially in recipient strain P. aeruginosa 280. This was shown to result in the failure to detect gentamicin resistance transfer if the concentration of gentamicin in selection media was too high (>2.5 mug/ml for strain 280). Some but not all recipients were changed in pyocine type upon acquisition of R-factors. PMID- 15828192 TI - Effect of chelating agents on the susceptibility of some strains of gram-negative bacteria to some antibacterial agents. AB - The effects of ethylenediaminetetraacetic acid and related chelating agents on the susceptibility of Pseudomonas aeruginosa and other strains of gram-negative bacteria to some beta-lactam antibiotics and some non-antibiotic antibacterial agents have been studied by two methods: (i) the determination of minimal inhibitory concentrations in the presence and absence of a chelating agent; and (ii) pretreatment with a chelating agent, followed by exposure for up to 120 min at 37 C to a beta-lactam drug in a nutrient medium or to the other type of antibacterial agent in a non-nutrient environment. The pretreatment technique gave the more significant results, especially with chlorhexidine, cetrimide, and benzalkonium chloride. Chelating agents that had previously been found to give a low order of activity as measured by effects on bacterial viability nevertheless appeared to induce some changes in the surface layers of the treated organisms, rendering them susceptible to sub-inhibitory concentrations of non-antibiotic drugs. PMID- 15828193 TI - Relationship between the structures of fatty acid amide derivatives and their antimicrobial activities. AB - The structure-activity relationships of derivatives of the antibiotic cerulenin were investigated by chemically modifying dodecanoic acid, its skeletal back bone. The dimethylamide derivatives were active against both gram-positive and negative bacteria, and fungi. Among the compounds having modified groups at positions C2 and C4, the most active were those with a carbonyl group at C4 and a double bond at C2. The dimethylamide and pyrrolidine amide derivatives of this structure type were the most active. Activity against bacteria and yeast increased with the number of carbon atoms in the skeleton, with the maximum activity being observed at C=12. No significant differences in activity against fungi were observed with change in chain length. PMID- 15828195 TI - Antimicrobial activity of the volatile oil of Nigella sativa Linneaus seeds. AB - The antimicrobial activity of the volatile oil of Nigella sativa Linneaus seeds was studied. The antimicrobial principle has been isolated, identified as thymohydroquinone, and found to be active against gram-positive bacteria and yeasts. PMID- 15828194 TI - Effect of the inhibition of protein synthesis on the Escherichia coli cell envelope. AB - The consequences for cell envelope integrity of Escherichia coli K-12 of the inhibition of protein synthesis by a variety of means have been examined. Protein synthesis was blocked by the antibiotics chloramphenicol and streptomycin, by amino acid starvation of an amino acid auxotroph, and by inactivation of temperature-sensitive aminoacyl transfer ribonucleic acid synthetase and ribosomal mutations. Closely similar morphological and physiological effects were found irrespective of the means by which protein synthesis was blocked. Scanning electron microscopy revealed a spectrum of changes after protein inhibition, with granular material derived from cells and spheroplasts commonly seen. Streptomycin caused additional changes manifested in a collapsed appearance of treated cells. Measurements of the release of lipopolysaccharide from the cell surface, alterations in outer membrane penetrability, and lysis of lysozyme ethylenediaminetetraacetic acid-treated cultures also showed that the various inhibitory treatments all had similar effects on cell envelope properties. The close correspondence between the effects seen with antibiotic-treated cultures and those in which protein synthesis inhibition was achieved by use of mutants indicates that the effects of chloramphenicol and streptomycin on the cell envelope are indirect consequences of ribosomal block, rather than due to multiple sites of action of the antibiotics. PMID- 15828196 TI - Introduction--a general survey of human dietary change in Europe. PMID- 15828197 TI - Diet and the Neanderthals. AB - The ultimate goal of paleoanthropological studies is to develop the most accurate and exhaustive portraits possible of our extinct human relatives, and of the history by which we became what we are. This endeavor includes, in the first place, the essential processes of establishing the basic parameters of hominid diversity, and of elucidating the potential evolutionary relationships among the components of that diversity. But our efforts clearly need to go farther than this; for the overall picture of human evolution is quite evidently incomplete without consideration of early hominid lifeways, and of how now-vanished hominid species interacted with their environments. Among the most important interactions of this kind is, unquestionably, feeding behavior and the expression of such behaviors in diet. For, at least short of breathing, feeding is the most fundamental of all the subsistence activities in which a terrestrial species can engage. And we will never be able to claim to understand the lifeways of ancient hominids without at least some insight into how they sustained themselves. Self evident as these remarks might be, however, they should not be taken to imply that--especially among eurytopes such as hominids--diet can, or should ever be regarded as, monolithic, or as an intrinsic property of any species. Nor do they mean that we can ever look upon hominid populations as "adapted" to particular food resources. Indeed, primates in general are remarkably varied in the diets that may be chosen by different populations of the same species, both seasonally and geographically [see, for example, the review of dietary variation among Malagasy strepsirhines in Tattersall, 1982]. Rather, amongst most if not all living primates that have been studied, it appears--not surprisingly--that the factor, which most importantly controls immediate dietary intake, is the spectrum of potential food resources available within the local environment. Not to put too fine a point on it, most primates are opportunists. PMID- 15828198 TI - Menu of Gravettian people from southern Moravia. AB - There are a number of Upper Palaeolithic sites of Gravettian people in the southern Moravia. These people had eaten animals and their bones were used for creating artefacts. Their food was based on several species that lived in the vicinity of their settlement unit. The sites Dolni Vestonice II (Under Western Slope--UWS), IIa and III and Pavlov (1952, 1953, 1957 and 1958) have been studied to obtain a picture of the menu of Gravettian people in this region. Hunted animals fall into two groups, the first one includes those species hunted consistently and the second group those hunted occasionally. The following animals rank among the first group: mammoth, reindeer, horse, wolf, hare and fox. The second group includes bear, lion, wolverine, wildcat, lynx, deer, woolly rhinoceros and birds. The carnivores were hunted for their hides, fur and bones. The long bones of hunted animals were crushed for marrow. The proximal parts of bones were used for creating tools since distal parts of bones have been found predominantly. Teams of several hunters hunted herd animals. The rest of the species was hunted accidentally, some of them probably by hunting nets. PMID- 15828199 TI - A re-evaluation of localized hypoplasia of the primary canine as a marker of craniofacial osteopenia in European Upper Paleolithic infants. AB - Localized hypoplasia of the primary canine (LHPC) occurs in prehistoric and contemporary populations with prevalence varying from 0 to 89%. One of the highest prevalences ever reported is Upper Paleolithic infants from Europe where 70% are affected. In that LHPC is found in relatively high proportions of contemporary children with reported or suspected malnutrition, it is important to investigate the etiology of LHPC. Previous research indicates that LHPC occurs in two steps: craniofacial osteopenia results in temporary fenestration of the cortical bone overlying the primary canine crypt; secondly, minor physical trauma to the perioral region impacts on the unprotected forming tooth crown resulting in a small pit visible on the labial surface of the erupted tooth. Investigation of the prenatal diet of mothers whose children are shown later to have LHPC found mothers are significantly low in vitamin A. Hitherto an animal model for this problem has been lacking. Recently Newell and Skinner have recognized that LHPC occurs very commonly in orangutans. Recent study of infant jaws from Pongo pygmaeus (n=75) and Pan paniscus (n=39) shows all stages of fenestration and healing of the labial bone of the primary canine crypt. 85% of orangutan and 62% of bonobos show LHPC. Current research is directed at bioavailability of vitamin A to ape infants. LHPC is a marker of malnutrition; a common factor that could link orangutan and Upper Paleolithic mothers is low dietary fat intake. A minimal level of dietary fat is required for gut absorption of both vitamin A and carotenoids. PMID- 15828200 TI - Trace elements and the European skeleton through 5000 years. AB - For our research, one thousand forty-four samples were taken from the femurs of 522 skeletons from 25 sites in Europe from the Neolithic Age (4000-5000 B.C), from La Tene Period, the Roman Era (500 B.C-400 A.D.), the Middle Ages and from contemporary cadavers. We found the following distribution of elements in the longitudional axis of long bones (the femurs and the tibias). The elements Zn, Fe, Ni, Cr, Pb, Mn, Co and Sn cumulated in the epiphysis. On the other hand, we found that Ca, Sr, Na and K prevailed in the central part of the diaphysis. In the central parts of the cross-section the highest concentration of the metal element Pb was in the external layer from the historical femurs. It was the same with cadavers of the recent population. A specific shift from Neolithic farming to agricultural intensification in the Roman Era was also apparent in the skeletons. Special sources of the above mentioned elements were found both in Celtic and Germanic tribes. Meat is the main source of zinc. Zinc is also important for the growth of the skeleton. When we investigated the development of the human skeleton during the last 5000 years we found the highest concetrations of Zn in communities with a good supply of animal food, whereas the lowest concentrations were paralleled with well-developed agriculturists. We assume the Neolithic gracilization, which is in the background for the increase of agricultural populations, is directly linked with the concentration of Zn and other elements essential for growth (Cu, Fe and others). The individuals most vulnerable to zinc deficiency include infants, adolescents during rapid growth phases and women during pregnancy and lactation. Trace elements in the bones of the La Tene period designate two areas of Celtic diet patterns--a "French one" (Roulier, Mont Trote and Acy Romance) and a "Czech one" (Karlov, Radovesice and Jenisuv Ujezd). At Czech sites levels of zinc increased westward towards the Germanic region. Over the Germanic territory in the region of the Saala River there are similar supply trace element sources for bones of the Germanic tribes for a period of more than 1000 years. The dietary customs and environment that formed this development were preserved from 400 B.C. to the period of the Merovingians. A specific ratio of Zn and Sr can be found not only in men, but also in woman and children. At the beginning of our era lead emerged as a civilization element. This element influenced the diet until the 20th century. Since the beginning of the 20th century it occurs much less in the population. On the contrary, we have found significant presence of tin in human bones. Cadavers of the modem population indicate a high content of tin, considerably higher than those in the populations from the beginning of our era (as much as 40 microg.g( 1) bone). PMID- 15828201 TI - Archaeometallurgy of the Etruscan dental protheses: prestige, magic or biocompatibility? PMID- 15828202 TI - Diet reconstruction in the Roman era. AB - The samples from the proximal femora were taken from 12 cementeries from the Roman period. The skeletons date from the 1st-4th centuries A.D. Trace element analysis was used in order to reconstruct the basic diet. The sites that best corresponded to the model of Old Germanic diet described by ancient authors "meat, milk and cheese" were found in the Pruszcz Gdanski East Pomerania region close to the Baltic sea as well as in region Halle (Niemberg) and not far from Donau (Sladkovicovo). This diet is characterized by a large amount of protein and consequently of zinc. In the original Old Germanic region in a time period of more than 1000 years (from 400 B.C. to 700 A.D.) there is the same type of trace elements sources for bones and also the same type of the diet. This possibly distinguishes Germanic soldiers (aboriginal) from others groups in Roman legionary camps. It seems that the Donau River is very important for predicting the type of diet in the Roman period. North of the river animal component prevails south of the river vegetal component prevails. The rich agricultural land along the Donau River and in the Pannonian plains affects social arrangement as well as the structure of bones in Germans and Sarmats in the 2nd-4th centuries. Lead became a civilization element. It appeared in the diet of the Greek and Romans. Contamination varied with different social classes. We have found higher lead concentration in the femurs of the Germans than in those of the Sarmatians. The highest concentration we found was in Pannonian towns (Gorsium, Sopianae) and legionary camps (Straubing, Gerulata). In reference to age, the maxima of the highest lead concentrations in Gerulata II are between 11 and 13 years of age and between 40 and 50 years. PMID- 15828203 TI - The Longobard population from the San Mauro Necropolis Cividale: preliminary results of our anthropological and palaeonutritional study. PMID- 15828204 TI - A singular case of dwarfism with articular facets on tibio-fibular diaphysis, from the Longobard necropolis of the late-middle-ages at Cividale del Friuli (Udine)--Italy. PMID- 15828205 TI - Cribra orbitalia in early medieval population from Ostrow Lednicki (Poland). AB - Cribra orbitalia is mostly diagnosed as being of nutritional origin, especially iron deficiency anaemia, and as the effect of infectious diseases and parasitic infestations. That lesion of the orbital roof is the good indicator of the biological state of the prehistoric populations. The frequency of occurrence of the cribra orbitalia was examined in the early medieval (12th-14th century) human population from Ostrow Lednicki (an island on the Lednica Lake--Wielkopolska Province, Poland). The intensity of the cribra orbitalia was estimated on the basis of Hengen's scale, and author's scale (3 degrees of intensity). 494 adult skulls, 259 females (52.4%), 234 males (47.4%) and one indeterminate individual (0.2%) were analysed. Cribra orbitalia were present in 149 (30.2%) individuals of this population. Frequency of occurrence of this lesion was higher in females (17.4%) than in males (12.8%), and its distribution in different age categories was: adultus 15.8% (78 individuals), maturus 12.6% (62 individuals), senilis 1.8% (9 individuals). Cribra orbitalia were more frequent in the left orbit. Occurrence and intensity of this pathology of the orbital roof was compared with the various osteometric features: measurements of the skull, calculated body height and massivity of the postcranial skeleton. There was lack of the correlation among those features and cribra orbitalia. Results of this study were compared with other medieval skeletal series from Poland. PMID- 15828206 TI - Trace elements and heavy metals as indicators of palaeodiet and dating of human population. PMID- 15828208 TI - Europe's great dietary revolution: from a single staple (bread) to a diversity of food, 1750-1900. PMID- 15828207 TI - Assortment of the plants in the Medieval diet in Czech countries (based on archaeobotanical finds). AB - The finds of remains of useful plants in archaeological objects were drawing attention already in the last century. However, it was not until after the World War II, in relation with renovation of historical cores of the towns, when a separate branch of the botany--archaeobotany constituted. In the Czech Republic it was E. Opravil who was the first to become engaged in a systematic archaeobotanical research at the beginning of the 60's. All the vegetal remains- fruits and seeds on the first place as well as pollen grains from prehistoric times to the post-medieval period, are the subjects of archaeobotany. The medieval waste pits are the richest sources of the vegetal remains. They contain a lot of diasporas of cultural as well as wild plants. The remains of useful plants are an authentic document of the vegetal component of diet. Even though some vegetal species, such as for example garlic, onion or stalk-plants, about whose cultivation we are informed from literature, occur only seldom, there appeared also the species not mentioned in literature. All the basic cereals have been known in our country already since the early Middle Ages. Wheat (Triticum aestivum) and rye (Secale cereale), as well as millet (Panicum miliaceum) that, along with buckwheat (Fagopyrum esculentum), were replaced by potatoes in the modern times were the most important. Also leguminous plants were of great importance in the diet even though their charred seeds seldom preserved. Remains of oil plants such as hemp (Cannabis sativa), flax (Linum usitatissimum) and poppy (Papaver somniferum) preserved well. It was successfully proved that cucumber (Cucumis sativus), among other numerous kinds of vegetables, was introduced by the Slavs already in the 8th century in Moravia and in the 9th century in Bohemia. The assortment of fruit was rich--especially the assortment of cultivated fruit. Among the imported kinds of fruit the first place occupied figs (Ficus carica), from the numerous finds it can be said that they formed a common part of the diet. Among imported fruit and spices from the peak period of the Middle Ages we succeeded in finding a date-tree (Phoenix dactylifera), nutmeg tree (Myristica fragrans) and, by means of pollen analysis, maybe a clove tree (Eugenia caryophyllata). PMID- 15828209 TI - Changing circumstances for accepting of behavioral patterns in connection with food preparation. AB - Food habits in a family are the results of many factors, e.g. the local traditions, availability of food, economic, social and cultural situation, and many others depending on the particular environment. Key role in this respect in the family have women, especially mothers. The transmission of the experiences on the choice and preparation of daily meals, their characteristic, frequency of their intake is also a result of previous generation of mothers, i.e. grandmothers. However, this situation has started to vary during more recent period of time according to the global changes in lifestyle. This resulted especially from the economic development and changing social situation. The analysis of this situation, when the role of mother and family declines, in the transmission of food choice and habits will be presented. PMID- 15828210 TI - How will disclosure affect future litigation? PMID- 15828211 TI - Disclosing medical error: how much to tell? PMID- 15828212 TI - Recognizing and preventing hypoxemic injury risk on the general care floor. PMID- 15828213 TI - Risk management implications of increasing PDA usage doctors. PMID- 15828214 TI - Vicarious liability. Independent medical staff member may be agent of hospital. McCorry v. Evangelical Hospital Corporation. PMID- 15828215 TI - Professional liability. Anesthesiologist who started chain of events leading to injury may be liable. HealthONE v. Rodriguez. PMID- 15828216 TI - Informed consent. Material misrepresentation of qualifications may invalidate informed consent. Howard v. University of Medicine and Dentistry of New Jersey. PMID- 15828217 TI - [Mechanism for the action of PPARs]. PMID- 15828218 TI - [Recent advances in PPARs research--basic and clinical aspects]. AB - PPARs (peroxisome proliferator-activated receptors), PPARalpha, beta/delta and gamma, play crucial roles in regulation of glucose and lipid metabolism, energy homeostasis and atherosclerosis. Specific and common actions of these three PPARs have recently been elucidated using mice models and specific, dual, or pan agonists. A number of drugs targeted to PPARs are being developed for metabolic disorders such as diabetes and hyperlipidemia and atherosclerosis diseases. This review focuses the recent advances in PPARs research from basic and clinical aspects. PMID- 15828219 TI - [PPARs as molecular targets for drug discovery]. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-inducible transcription factors. PPARs form the heterodimers with retinoid X receptors (RXRs), and modulate specific gene expressions related to the lipid and carbohydrate metabolism. Since fibrate-type lipid-lowering agents and thiazolidinedione (TZD)-type antidiabetic agents are identified as PPARalpha and gamma agonists, respectively, a number of ligands for PPARs have been developed, including PPAR subtype-selective and dual agonists. Recent report that RXR antagonist exhibited antidiabetic and antiobestic activities in animal model experiments, PPARs partial agonists, antagonists, and RXR antagonists have been also synthesized and their function in vivo are under investigations. In this paper, recent studies on the ligands for the PPAR-RXR heterodimers are reviewed. PMID- 15828220 TI - [The structures and functions of peroxisome proliferator-activated receptors (PPARs)]. AB - Peroxisome proliferator-activated receptor (PPAR) is a member of the nuclear receptor superfamily, which is activated by various hydrophobic compounds. PPAR has 3 subtypes (PPARalpha, gamma, and delta) showing different expression patterns. Each PPAR subtype has homologous domains (DNA binding domain and ligand binding domain). PPARalpha plays a role in the clearance of circulating or cellular lipids via the regulation of gene expression involved in lipid metabolism in liver and skeletal muscle. PPARgamma promotes adipocyte differentiation to enhance blood glucose uptake. PPARdelta is involved in lipid oxidation and cell proliferation. Therefore, all PPAR subtypes are indispensable for the regulation of wholebody lipid metabolism. PMID- 15828221 TI - [PPARs-mediated intracellular signal transduction]. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that are activated by fatty acids and their derivatives. PPARs consist of three isotypes named PPARalpha (NR1C1), PPARbeta/delta (NR1C2) and PPARgamma (NR1C3) in vertebrates. Each of them is encoded in a separate gene and binds fatty acids and eicosanoids. Although each isotype fulfills distinct functions, PPARs function not only as an important fatty acid sensor that regulate lipid, carbohydrate and amino acid metabolism but also play an important role in various signaling pathways (immunity, inflammation, apoptosis and cell differentiation). In this review, I focus on emerging knowledge about PPARs roles in various intracellular signaling pathways. PMID- 15828222 TI - [PPARs target genes]. AB - PPAR(alpha, beta/delta, gamma) are ligand-dependent nuclear receptors and regulate homeostasis, cell proliferation/differentiation and associate with hypolipidemia, atherosclerosis, diabetes, and obesity. Through heterodimerization with retinoid X receptors (RXRs), PPARs bind the same consensus response element, formed by a direct repeat of two AGGTCA hexamers separated by one base. Recently, many PPAR direct and indirect target genes have been reported. Here, we summarize the PPAR direct/indirect target genes, and their functions related to lipid metabolism, adipocyte differentiation. PMID- 15828223 TI - [Endogenous ligands for PPARs]. AB - Peroxisome proliferator-activated receptors (PPARs), a family of three nuclear receptors/transcription factors, are widely recognized as molecular targets for drugs against lifestyle related disease. In spite of intensive search for natural ligands, no truly endogenous ligand has been identified as yet. Rather, these results have lead to the suggestion that PPAR may act as various lipid sensors. Namely, the ligand binding modes of PPARs would be similar to those of odorant receptors or substrate-binding modes of drug-metabolizing enzyme P450 family. In this brief review, free fatty acids, lipid mediators in arachidonate cascade and polyphenolic compounds such as resveratrol will be discussed as natural ligands for PPARs. PMID- 15828224 TI - [Roles of PPARs in carcinogenesis]. AB - Peroxisome proliferator activated receptors (PPARs) is known to be expressed in several cancers, but their roles on carcinogenesis has not been confirmed. PPARalpha and delta have not been defined whether as an accelerator or an inhibitor, but now PPARgamma is thought as an inhibitor on carcinogenesis. Therefore, we attempted to elucidate the role of PPARgamma in gastric carcinogenesis and explore the possibility of using PPARgamma ligand as chemopreventive agent for gastric cancer with PPARgamma knockout mice. The present study demonstrated that PPARgamma suppresses gastric carcinogenesis, the PPARgamma ligand troglitazone is a potential chemopreventive agent for gastric carcinogenesis, and troglitazone's chemopreventive effect is dependent on PPARgamma. Unfortunately, thiazolidinediones such as troglitazone has side effects, including edema at the high dose of administration. So we expect the development of new PPARgamma ligands, which are more selective and powerful for PPARgamma, to break through the difficulties of PPARgamma ligands' use clinically. PMID- 15828225 TI - [Roles of PPAR family in bone metabolisms]. AB - Bone is a complex tissue which contains osteoclasts, osteoblasts, chondrocytes, adipocytes, hematopoietic cells and immune cells. Since osteoblasts share the same origin with adipocytes in bone marrow cavity, it is assumed that PPAR (peroxisome proliferator-activated receptor) family, which is an important nuclear receptor family for adipocyte differentiation, plays a role in the bone microenvironment. Indeed, recent evidences support the primitive roles of PPAR family in osteoblast differentiation as well as adipocyte differentiation. Furthermore, PPAR family is also implicated in the regulation of differentiation and function of osteoclasts. Here, we summarized the functional roles of PPAR family in bone remodeling and regulation of bone microenvironments. We also discuss the potential mechanisms that regulate expression and function of PPAR family during bone metabolisms. PMID- 15828226 TI - [PPARs and fibrosis]. AB - Inflammation is associated with fibrosis. Angiotensin II-stimulated growth of fibroblasts and an increase in collagen type I synthesis are important component of the cardiac remodeling process in hypertension and chronic ischemia. AngII has been shown to enhance production of reactive oxygen species (ROS) via stimulation of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase. Recent studies have proposed that stimulation of ROS production by AngII may constitute a means by which this humoral factor contributes to development of tissue injury in organs such as blood vessels, kidney, and the heart. Published studies have shown that PPARgamma ligands can attenuate the expression or activity of NADPH oxidase subunits. Furthermore, it has been shown that PPARs inhibits inflammation by blocking the activation of redox-sensitive transcription factor NFkappaB. Although there is much still to learn about the link of inflammation and fibrosis, PPARs are potential therapeutic targets for treating cardiac fibrosis and perivascular fibrosis. PMID- 15828227 TI - [Peroxisome proliferator activated receptor ligands and angiogenesis]. AB - PPAR receptors are expressed in endothelial cells and PPARalpha and gamma ligands play potent roles in modulating angiogenesis. Angiogenesis, the formation of new capillary blood vessels, is essential for many physiologic and pathologic processes such as cancer, choroidal neovascularization and ischemic diseases. Inhibition of angiogenesis can prevent diseases with excessive vessel growth such as cancer or diabetic retinopathy. Vascular endothelial growth factor (VEGF) and VEGF-receptors are considered key regulators of angiogenesis. PPARalpha and gamma ligands not only inhibit VEGF-receptor 2 expression but also act as blocker of VEGF-receptor signaling. Antiangiogenic action through targeting of VEGF receptors may be a new approach to blocking hypoxia-driven angiogenesis. In this review, I highlight data concerning PPARs ligands involved in the biological mechanisms underlying antiangiogenesis. PMID- 15828228 TI - [Anti-inflammatory action of PPARs]. AB - The PPARs transduce a wide variety of signals, including environmental and nutritional events, into a defined and ordered set of cellular responses at the transcriptional level. It has now been shown that all three PPAR isoforms (alpha, beta/delta and gamma) can participate in the regulation of inflammation responses. The ability of the PPARs to regulate inflammatory responses is a result of their transactivation and transrepression capacities. Most of the anti inflammatory properties of the PPARs arise through their ability to antagonize nuclear factor-kappaB (NF-kappaB) and AP1 signaling pathways. By inhibiting these inflammatory transcription factors, the PPARs repress the expression of several genes that are involved in the inflammatory responses. In this review, I focus on emerging knowledge about PPARs roles in inflammatory responses. PMID- 15828229 TI - [Polymorphisms of PPARs and metabolic disorders]. AB - Peroxisome proliferator-activated receptors (PPARs) are a member of transcriptional factor that regulates expression of several genes associated with glucose and lipid metabolisms. Some polymorphisms of PPARs gene have been reported, and the impacts of these polymorphisms are examined in patients with several metabolic disorders. The Pro12Ala polymorphism of the PPARgamma gene is studied extensively, and is known to be associated with increased insulin sensitivity and reduced risk for the development of diabetes. Such studies will help us to elucidate the role of PPARs on the development of metabolic disorders and to find new therapeutic approaches to these disorders. PMID- 15828230 TI - [PPAR and diabetes]. AB - The peroxisome proliferator-activated receptors (PPARs) are nuclear fatty acid receptors that have been implicated to play crucial roles in metabolic diseases such as hyperlipidemia, insulin resistance, and diabetes. The three PPAR subtypes, alpha, gamma, and delta, have distinct expression patterns and regulate glucose homeostasis based on the need of a specific tissue. While PPARalpha potentiates fatty acid catabolism in the liver and is the molecular target of the lipid-lowering fibrates, PPARgamma is essential for adipocyte differentiation and hypertrophy, and mediates the activity of the insulin sensitizing thiazolidinediones. PPARdelta may be important in regulating body weight and lipid metabolism in fat tissues. Thus, uncovering the regulatory mechanisms and transcriptional targets of these PPARs will provide insights into the pathogenesis of metabolic diseases and offer valuable information for rational drug design. PMID- 15828231 TI - [PPARs and hypertension]. AB - PPARs (peroxisome proliferator-activated receptors) alpha, delta and gamma are expressed in adipocytes, skeletal myocytes and hepatocytes and also in endothelial cells, vascular smooth muscle cells and monocytes/macrophages, in distinct distributions. They are critically involved in the pathophysiology of obesity, diabetes, hyperlipidemia and hypertension, the complex of which is recognized as "metabolic syndrome", as well as, of their vascular complications. In addition to modulation of insulin sensitivity, PPARs can regulate oxidative stress and local production of various vasoactive substances in blood vessels, and thus control vascular tone and remodeling as one of the determinants of hypertension. PMID- 15828232 TI - [Hyperlipidemia and peroxisome proliferator-activated receptor (PPAR)--regulation of the PPARalpha gene by CLOCK: BMAL1]. AB - The main pathological findings in atherosclerosis include abnormal reactions of neutrophils, lymphocytes and monocytes/macrophages, vascular smooth muscle cells and vascular endothelial cells, and the accumulation of cholesterol ester in the arterial wall. Therefore, investigating the effects of these abnormal reactions on the arterial wall may improve understanding of the mechanisms underlying atherosclerosis. Three types of peroxisome proliferator-activated receptors (PPARs): PPARalpha, PPARbeta/delta, and PPARgamma are expressed in endothelial cells. In endothelial cells, the ligands/activators for PPARalpha and PPARgamma increase Cu2+, Zn2+ -superoxide dismutase. In addition, the phorbol myristate acetate (PMA)-stimulated 22 kDa-subunit (p22phox) protein levels and 47 kDa subunit (p47phox) protein levels in NADPH (superoxide generating enzyme nicotinamide adenine dinucleotide phosphate (reduced form)) oxidase were decreased by treatment with PPARalpha and PPARgamma ligands/activators. Recently, we showed that the CLOCK: BMAL1 heterodimer regulates the PPARalpha gene via promoter of PPARalpha. Moreover, we report a patient with severe hypertriglyceridemia associated with anemia and hypoalbuminemia, in which the former may have caused the latter two conditions. This is the first reported case of abrupt onset of severe hypertriglyceridemia resulting in suppression of bone marrow and liver function. Here, based on recent studies including our own, we describe the relationships between risk factors for atherosclerosis, especially hyperlipidemia and PPARs and the molecular mechanisms that govern lipid metabolism in the arteries. PMID- 15828233 TI - [PPARs in atherosclerosis]. AB - Atherosclerosis is considered as a chronic inflammatory condition. PPARs are also thought to be involved in the control of inflammatory response, which may suggest an additional role in the development of atherosclerosis. PMID- 15828234 TI - [PPARs and heart diseases]. AB - After it has been reported that activation of PPARalpha or PPARgamma suppresses production of proinflammatory cytokines, medical interest in PPARs have grown and a huge research effort has been concentrated. Synthetic antidiabetic thiazolidinediones (TZDs) and natural prostaglandin D2 (PGD2) metabolite, 15d PGJ2, are well known as ligands for PPARgamma. Hyperlipidemic drugs (fibrates) are synthetic PPARalpha ligands. Recent studies suggest that ligands of PPAR not only regulate glucose and lipid metabolism but also have pleiotropic effects on many tissues and cell types. Ligands of PPARs may become therapeutic agents useful in the prevention of cardiovascular diseases beyond their effects on glucose and lipid metabolism. This review will focus on the latest developments in the PPARs field and the roles of PPAR-dependent pathway in cardiovascular diseases. PMID- 15828235 TI - [The roles of PPARs in digestive diseases]. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), one of the nuclear receptors, is expressed at high levels in adipose tissue and is related to adipocyte differentiation and insulin sensitivity. PPARgamma is also expressed at high levels in digestive organs, especially in the colon epithelium. The physiological and pathological roles of PPARgamma in the gastrointestinal tract have been investigated and recognized as an endogenous regulator of innate immunity, inflammation, and cell proliferation. PPARgamma ligands have recently been reported to improve the condition of patients with non-alcoholic steatohepatitis (NASH). Thus, PPARgamma ligands are indeed potential benefit candidates for various digestive diseases. In this review, we will focus on the current knowledge and new insights of the roles of PPARgamma in digestive diseases, especially in (1) inflammatory bowel disease (IBD), (2) colon cancer, and (3) NASH. PMID- 15828236 TI - [Relationship between peroxisome proliferator activator-receptor (PPAR) and urological cancer]. AB - Recent studies have demonstrated that peroxisome proliferator activator-receptor (PPAR)-gamma ligand induces growth arrest of cancer cells through apoptosis. In this review, we demonstrated the expression of PPAR-gamma in urological cancer (renal cell carcinoma, bladder tumor, prostate cancer, and testicular tumor) tissues as well as the effects of PPAR-gamma ligands. While no marked expression of PPAR-gamma was observed in normal tissue, significant expression was found in all urological cancer tissue. PPAR-gamma ligands (troglitazone and 15dPGJ2) induced the reduction of cell viability with the half-maximal concentration of growth inhibition of renal cell carcinoma, bladder tumor, and prostate cancer cell lines. Cells treated with PPAR-gamma inhibitors showed typical redundant characteristics of apoptosis. PPAR-gamma ligands may mediate potent antiproliferative effects through differentiation. PMID- 15828237 TI - [Polycystic ovary syndrome]. AB - Polycystic ovary syndrome (PCOS) is an endocrine disorder characterized by chronic unovulation, hyperandrogenism and polycystic change in ovary. Hyperinsulinemia is so often accompanied with PCOS that insulin resistance may play important roles in pathogenesis of PCOS. Recent studies reported the effectiveness of insulin-sensitizing drugs on treatment of patients with PCOS. Thiazolidinedione, an agonist of PPARgamma receptor, improves not only insulin sensitivity but also hyperandrogenism and ovulatory dysfunction in patients with PCOS. Insulin-sensitizing drugs such as thiazolidinediones are expected to be a novel therapy for PCOS, although further studies on the effectiveness and safety should be required. PMID- 15828238 TI - [Autoimmune disease]. AB - Peroxisome proliferator-activated receptors (PPARs) are members for the nuclear hormone receptors superfamily of ligand-activated transcriptional factors. PPARs have diverse function for keeping physiology of the human body, such as fatty acid regulation. In particular, PPAR-alpha and -gamma, have anti-inflammatory function and play an important role in immune system. With these ligands, we can suppress inflammation and regulate abnormal immune reaction in variety of immunocompetent cells and organs. In this review, we show the utility of PPARs ligands in models for autoimmune disease, and also examine that these ligands have the possibility of application for useful treatments of human autoimmune diseases. PMID- 15828239 TI - [Anti-hypertensive effects of PPARgamma ligands through the inhibition of Rho/Rho kinase pathway]. AB - Although PPARgamma ligands have an antihypertensive effect in vivo, the precise mechanism has not been fully elucidated. We examined their effects on Rho/Rho kinase pathway, a key regulator of vascular tone. In cultured rat aortic smooth muscle cells, Rho kinase activated by angiotensin II was suppressed by the pretreatment with pioglitazone and troglitazone. The roles of Vav, a GTP/GDP exchange factor upregulating Rho kinase activity, and SHP-2, protein tyrosine phosphatase that dephosphorylated Vav and subsequently inactivated Rho kinase were examined. Both pioglitazone and troglitazone upregulated SHP-2, particularly in the cytosolic fraction, and the SHP-2-bound Vav, and reduced the phosphorylation of Vav. These mechanisms may contribute to the hemodynamic, in addition to metabolic, action by PPARgamma ligands in hypertensive, metabolic syndrome. PMID- 15828240 TI - [Role of PPARs in the pathophysiology of nonalcoholoic fatty liver disease]. AB - Pathogenic role of intrahepatic lipid accumulation in insulin resistance and metabolic syndrome has been well documented. Liver steatosis constitutes a risk factor for nonalcoholic steatohepatitis (NASH), one of the leading causes of obesity-related morbidity and mortality. Although pathophysiology of steatosis is multifactorial, a line of evidence from rodent studies suggests that PPARalpha and PPARgamma are involved. PPARalpha is highly expressed in liver and its activation by agonists leads to augmented fatty acid oxidation and protects against steatosis. PPARgamma, which is transcriptionally up-regulated in steatosis, activates lipogenic enzymes and exacerbates steatosis. However, recent human studies have suggested that PPARgamma agonists improve NASH possibly by its primary insulin-sensitizing effect on adipocytes. PPARs modulation is becoming a rational and effective therapeutic approach for the treatment of nonalcoholic fatty liver disease. PMID- 15828241 TI - [SFRP, a family of new colorectal tumor suppressor candidate genes]. AB - Aberrant Wnt pathway signaling is an early progression event in up to 90% of colorectal cancers (CRC). It occurs mainly through mutations of APC or beta catenin. We have identified frequent promoter hypermethylation and gene silencing of the secreted frizzled-related protein (SFRP) genes in CRC. SFRPs possess a domain similar to one in the Wnt receptor Frizzled proteins and can inhibit Wnt receptor binding to downregulate pathway signaling in development. We next found that restoration of SFRP function in CRC cells attenuates Wnt signaling even in the presence of downstream mutations. We also show that the epigenetic loss of SFRP function occurs early in CRC progression and may, thus, provide constitutive Wnt signaling which is required to complement downstream mutations in the evolution of CRC. PMID- 15828242 TI - [Bone loss in men under androgen-deprivation therapy for prostate cancer]. AB - Bone is a major target tissue for sex hormones and hypogonadism is a known cause of male osteoporosis. Androgen-deprivation therapy (ADT) for prostate cancer yields hypogonadotropic status and accelerates age-related decrease in bone mineral density. The risk for osteoporotic fractures is also increased which may lead to a shorter life expectancy for prostatic cancer patients. Baseline and follow up bone density measurement is desired for all men beginning ADT and it is advisable to take calcium and vitamin D as well as maintain a moderate exercise. Bisphosphonate is a possible treatment for those in whom osteoporosis develops. More data are desirable to make a guideline to prevent and treat male osteoporosis under ADT. PMID- 15828243 TI - [Therapeutic strategies for postinfarction left ventricular free wall rupture]. AB - We treated 93 patients who developed left ventricular free wall rupture after acute myocardial infarction. Medical management including pericardial drainage was performed in 78 patients (84%), but 67 of them died. All 11 surviving patients showed an oozing type rupture. Surgical repair was performed in 15 patients (16%). As a result, 9 patients died and 6 survived. All but 1 of the patients who died presented with a blow-out rupture. Blow-out type rupture occurred in 3 and oozing type rupture in 3 of the surviving patients. One patient with blow-out type rupture underwent implantation of a left ventricular assist device following percutaneous cardiopulmonary support (PCPS), because of low output syndrome after the operation. The device was successfully removed 7 days after implantation. In all of the 3 patients with oozing type rupture, sutureless technique was successfully performed using fibrin-glue or fibrin-glue sheet fixation. After a mean follow-up period of 7 years after operation, 5 of 6 are still alive. To improve the clinical outcome of left ventricular free wall rupture, it is important for surgeons to closely liaise with physicians, to perform surgical repair as soon as possible, and to utilize a circulatory support system after operation. Therefore, we developed a new PCPS system compatible with emergency cardiac surgery and a new left ventricular assist system draining via the left ventricle. PMID- 15828244 TI - [Sutureless technique for oozing type postinfarction left ventricular free wall rupture]. AB - We report our experience using a sutureless technique for oozing type postinfarction left ventricular free wall rupture. Several materials such as fibrin seat, autologous or heterologous pericardial patch, fibrin glue, and geratin-resorcin-formaldehyde (GRF) glue have been used. Nine patients, who developed postinfarction left ventricular free wall rupture, underwent surgical repair using a sutureless technique between 1999 and 2004. All patients survived and discharged our hospital without any postoperative complications. And all are alive an exellent condition in 5 to 44 months. A sutureless technique for the treatment of oozing type postinfarction left ventricular free wall rupture is simple, effective, and associated with a favorable outcome. PMID- 15828245 TI - [Infarct exclusion for postinfarction left ventricular free wall rupture with severe congestive heart failure]. AB - A 70-year-old man was transferred to our hospital with severe congestive heart failure and ventricular arrhythmia due to acute myocardial infarction. He had experienced chest pain 3 weeks previously and was admitted to another hospital for dyspnea, where he required assist ventilation, 1 week prior to the transfer. An echocardiogram revealed a broad anteroseptal infarction and very poor left ventricular function with an ejection fraction (EF) of 22%. He remained in a severe congestive heart failure condition despite a full administration of catecholamines. Coronary angiogram findings revealed an occlusion of the proximal left anterior descending coronary artery and 1 week later severe hypotension was suddenly presented. An echocardiogram showed pericardial effusion with signs of cardiac tamponade. A pericardiocentesis was performed and hemodynamic improvement was obtained for a short time, after which the patient underwent urgent open heart surgery. During the operation, exclusion of the anteroseptal akinetic area using an oval patch was performed under a cardiopulmonary bypass and ventricular fibrillation. Severe cardiac failure remained postoperatively and the patient could not be weaned from cardiopulmonary bypass, therefore, we implanted a percutaneous cardiopulmonary support (PCPS) and started intraaortic balloon pumping (IABP). The patient was weaned from PCPS at 26 days after surgery and from IABP at 30 days. Following hospital release, he has continued to do well without heart failure for 39 months after the operation. PMID- 15828246 TI - [Risk factors for the surgical repair of ventricular septal perforation; an 8 year multiinstitutional analysis]. AB - OBJECTIVE: The purpose of this study was to determine the surgical outcomes and risk factors for surgical repair of the ventricular septal perforation (VSP). METHOD: From 1995 to 2003, 41 patients with VSP underwent surgical repair. There were 18 males and 23 females, with the mean age of 71.7 +/- 9.2. Sixteen patients (39.0%) had the preoperative shock, while 30 patients received intraaortic balloon pumping (IABP) assistance and 1 of those required percutaneous cardiopulmonary support (PCPS). Mean durations from onset of myocardial infarction and VSP to operation were 5.8 +/- 9.4 and 2.4 +/- 8.1 days, respectively. Twenty-six patients underwent infarct exclusion technique, 11 underwent patch closure, and 4 Daggett operation. Mean cardiopulmonary and aortic cross-clamp time were 211 +/- 85 and 105 +/- 43 minutes, respectively. RESULTS: Thirty days mortality was 11 (26.8%). Nine patients (22%) required PCPS after repair, however, 2 weaned off the support and only 1 discharged the hospital. Residual shunt was found in 12 patients (29.3%), and 4 underwent the reclosure of the residual shunt 13 +/- 8.6 days after the initial operation, whereas none of patients with PCPS had residual shunt. Univariate analysis revealed the preoperative shock (p = 0.03), longer cardiopulmonary bypass time (p < 0.01), and the need for PCPS after repair (p < 0.01) were the risk factors for the early mortality. Multivariate analysis indicated the cardiopulmonary time over 210 minutes and the need for PCPS to be the significant risk factors. CONCLUSION: The long cardiopulmonary bypass support after repair and the subsequent need for PCPS imply the poor left ventricular function. Since the residual shunt was not the cause of PCPS, the surgical outcome for VSP may be limited in patients with poor left ventricular function. In these patients, other therapeutic strategies may be required, such as ventricular assisting devices, transplantation, or regenerative therapy. PMID- 15828247 TI - [Surgical treatment for postinfarction ventricular septal perforation]. AB - Between November 1985 and April 2003, surgical treatment for ventricular septal perforation (VSP) after acute myocardial infarction was performed in 16 patients. Patients were divided into 2 groups by method of operation. One group is infarct exclusion technique (n = 10). The other group is trans right ventricular (RV) approach (n = 6). No significant differences were observed between 2 groups in preoperative states. Operative death rate was high in both groups. Five patients (50%) were died in infarct exclusion group, 4 patients (67%) in trans RV group. Infarct exclusion technique needed longer extracorporeal circulation time (201 +/ 33 min) than trans RV approach (170 +/- 32 min). Although trans RV approach is attractive for its simplicity, 3 cases died of rupture of remained left ventricle infarction area. On the other hand, there were no mortality cases caused by left ventricle rupture in infarct exclusion technique group. As this result, we select infarct exclusion technique as a surgical correction for VSP. PMID- 15828248 TI - [Modified Komeda-David operation for postinfarction ventricular septal perforation]. AB - The infarct exclusion technique with a xeno-pericardial patch which Komeda and associates firstly reported in 1990 is one of the best procedure to close ventricular septal perforation. A large patch can protect the perforation and the surrounding weak tissue from the internal left ventricular pressure. However, suturing this large patch to the left ventricular wall through the small incision is not technically easy because of the patch design in the ventricle. We modified the design of the patch. This round shaped bovine pericardial patch was sutured continuously to the left ventricular wall. Then, excessive part of the patch was trimmed to make a corn shape. Finally, the corn shaped patch fit the left ventricular wall naturally. PMID- 15828249 TI - [Surgical management of papillary muscle rupture following acute myocardial infarction]. AB - Papillary muscle rupture is a rare but severe complication of acute myocardial infarction. Two cases successfully underwent mitral valve replacement and concomitant coronary artery bypass grafting (CABG) for acute myocardial infarction with the anterior papillary muscle rupture in cardiogenic shock. Each of them needed preoperative massive inotropic infusion, respiratory support and intraaortic balloon pumping assist. The first case was a 76-year-old female. Double vessel disease (seg 7 : 90%, seg 11 : 100%) was revealed by coronary angiography and rupture of the papillary muscle was confirmed by transesophageal echocardiography. The second case was a 69-year-old female. Double vessel disease (seg 2 : 90%, seg 11 : 100%) was revealed and severe mitral regurgitation due to prolapse of the anterior leaflet was confirmed by transthoracic echocardiography. To assess the diagnosis of postinfarction papillary muscle rupture, transthoracic and/or transesophageal echocardiography is mandatory. Coronary angiography is also desirable because concomitant myocardial revascularization may improve the prognosis. PMID- 15828250 TI - [Overlapping ventriculoplasty for ischemic cardiomyopathy]. AB - We have previously reported overlapping cardiac volume reduction operation (OLCVR) for dilated cardiomyopathy. Because of the acceptable clinical outcome and especially the excellent ellipsoidal shape of the left ventricle (LV) after surgery, we extended this indication for ischemic cardiomyopathy (ICM) with dilated LV. In such cases we combined OLCVR with mitral annuloplasty and papillary muscles approximation (PMA), called integrated overlapping ventriculoplasty (IOLVP). From March 2003 to July 2004, we performed IOLVP with coronary artery bypass grafting (CABG) for 8 patients who were diagnosed ICM. There is no operative mortality. Pre- and postoperative hemodynamics data are follows: LV ejection fraction (LVEF) improved from 22.4 +/- 11.9 to 33.4 +/- 10.5%, LV end-diastolic volume index (LVEDVI) decreased from 155.5 +/- 26.5 to 93.7 +/- 13.5 ml/m2 and LV end-diastolic diameter (LVDd) diminished from 66.0 +/- 8.9 to 60.5 +/- 8.4mm. Mitral regurgitation changed from 2.6 +/- 0.8 to 0.1 +/- 0.2 degree. New York Heart Association (NYHA) functional class improved from 3.3 +/- 0.5 to 1.3 +/- 0.5. LV shape became ellipsoidal without akinesis lesion. IOLVP is considered as a good option for ICM with dilated left ventricle. PMID- 15828251 TI - [Repair of a subepicardial aneurysm after myocardial infarction in a patient with congestive heart failure]. AB - Subepicardial aneurysm of the left ventricle is a rare complication of myocardial infarction. We report a case of a 75-year-old woman found to have a subepicardial aneurysm 4 months after an acute myocardial infarction. The patient was admitted to our hospital with congestive heart failure, and transthoracic echocardiography and left ventriculography showed an aneurysm on the posterior wall of the left ventricle, 4.6 x 3.5 cm in size. We performed patch repair using a Hemashield graft, with coronary artery bypass grafting. She was discharged on postoperative day 69, and has been well since. PMID- 15828252 TI - [Dor operation for left ventricular aneurysm with sustained ventricular tachycardia]. AB - A 60-year-old woman was admitted to our hospital because of syncope attack due to sustained ventricular tachycardia (VT). She was treated medically after cardiopulmonary resuscitation. Coronary arteriography revealed a 99% stenosis of right coronary artery (posterior descending artery: # 4 PD), a 90% stenosis of left descending artery (# 6) and left akinetic aneurysm was demonstrated. The patient successfully underwent Dor operation with endocardial cryoablation. The postoperative course was uneventful and the recurrence of VT was never recognized clinically. PMID- 15828253 TI - [Nifekalant hydrochloride as an effective treatment for postoperative ischemic heart disease]. AB - We experienced 2 effective cases of nifekalant hydrochloride. One patient was 76 year-old female who underwent emergent coronary artery bypass grafting (CABG) because of unstable angina pectoris (AP) and ventricular fibrillation (Vf). Her cardiac function had been decreased preoperatively due to old myocardial infarction (OMI). One day after CABG, she revealed sustained ventricular tachycardia (VT) and Vf. Although administrations of neither lidocaine hydrochloride nor magnesium sulfate were effective, nifekalant hydrochloride finally stopped the life-threatening arrhythmia without hypotension. Another patient was 77-year-old male who underwent CABG and Dor operation. His cardiac function also had been decreased due to OMI. He revealed VT attack at midnight 3 days after operation. VT attack still appeared at next 2 midnight under lidocaine hydrochloride infusion, but finally it has disappeared after starting a drip infusion of nifekalant hydrochloride. Nifekalant hydrochloride is quite useful as a new therapeutic strategy for uncontrollable VT and Vf and for the patient who has a reduced left ventricular function because it has an inotropic effect. PMID- 15828254 TI - [Complete resection for giant thymic carcinoma after simultaneous combination chemotherapy]. AB - A 19-year-old man visited our hospital complaining of dyspnea. Chest X-ray and computed tomography (CT) showed a huge mass in the right anterior mediastinum. We diagnosed this as invasive thymoma by microscopic examination of specimens obtained by echo-guided needle biopsy. The patient underwent 6 courses chemotherapy [1st course : carboplatin (CBDCA) + doxorubicin hydrochloride (DXR) + vincristine sulfate (VCR) + cyclophosphamide (CPA), 2nd, 3rd-6th course : cisplatin (CDDP) + ADM + VCR + CPA]. At achievement of partial response (the reduction rate of the tumor size : 91.4%), the tumor was completely resected. The pathological examination of the resected specimens yielded a diagnosis of large cell carcinoma. Preoperative chemotherapy with ADOC regimen may be effective in advanced thymic carcinoma. PMID- 15828255 TI - [Aortic valve replacement presence of anti-Jr(a) antibody]. AB - A 65-year-old female with a heart murmur developed progressive symptom of chest oppression. She was diagnosed severe aortic valve stenosis with echocardiogram. Antibody screening revealed anti-Jr(a) antibody. Preoperatively, erythropoietin was administered. Over 14 days, a total 1,000 ml of her blood was drawn and stored for autologous transfusion. The aortic valve was replaced with ATS mechanical valve [18 mm advanced performance (AP)]. Following surgery, her stored blood was administered to him. But her HCT was 17% on the 1st postoperative day. Frozen thawed red cells were transferred 7th postoperative day. PMID- 15828256 TI - [Left-sided adult Bochdalek hernia with right lung cancer; report of a case]. AB - Bochdalek hernia is a common congenital anomaly in neonatal patients with risky respiratory distress and high mortarity, but can be seen in adults. A case of left-sided adult Bochdalek hernia with right lung cancer is reported. A 71-year old female had been performed radition therapy for lung cancer in the right lower lobe. She was admitted to our hospital due to advanced lung cancer and pneumonia. On the 7th day after admission, she felt dyspnea and abdominal distention due to herniation of the stomach through the posterolateral defect of the diaphragma into the left hemithorax. Her condition did not allow us a radical surgery of Bochdalek hernia, so that we performed a palliative surgery, that is reduction of the stomach and gastrostomy. After surgery, her respiratory distress was lightened and she came to be able to ingest. She was less uncomfortable until she died due to progression of the lung cancer. PMID- 15828257 TI - [Right delayed traumatic diaphragmatic hernia; report of a case]. AB - An 80-year-old woman was admitted to our hospital with abnormal shadow on chest X ray 8 years after a chest trauma during which multiple ribs on the right side were fractured causing hemothorax. A diagnosis of right delayed traumatic diaphragmatic hernia was based on the findings on plain X-ray and multislice computed tomography. We performed surgery via thoracic approach with thoracoscopic assist. The transverse colon, liver, and omentum were dislocated into the right thoracic cavity and hardly adhered to the lung. We successfully repaired the ruptured diaphragm. The postoperative course was uneventful and the patient was discharged on the 33rd postoperative day. PMID- 15828258 TI - [Primary neurogenous sarcoma of the lung; report of a case]. AB - We report a case of a 27-year-old woman with primary neurogenous sarcoma of the lung. She had no symptoms but an abnormal shadow of the right lower lung field on the chest X-ray. Chest computed tomography (CT) revealed a well defined round mass, 20 mm in maximum diameter, at the right S9. Pathological study of the specimen obtained by CT-guided percutaneous needle biopsy showed undefferentiated carcinoma. Positron emission tomography (PET) disclosed intensely increased uptake of fluoro-2-deoxy-D-glucose (FDG) at the lung lesion without other abnormal uptakes. The patient underwent right lower lobectomy of the lung and mediastinal lymph nodes dissection. Results from immunohistological study yielded a definitive diagnosis of neurogenous sarcoma. Postoperative course was uneventful, and there has been no evidence of recurrence and metastasis for more than a year after the surgery. Reported cases of primary neurogenous sarcoma of the lung are reviewed. PMID- 15828259 TI - [Sternal metastasis of breast cancer; report of a case]. AB - A 51-year-old woman presented with a painful sternal lump. Nine years earlier she had undergone right modified mastectomy for breast cancer, followed by chemotherapy. Computed tomography and bone scintigraphy demonstrated a solitary sternal bone metastasis. The patient was treated by partial sternectomy. Thereafter, the sternal defect was reconstructed with Marlex mesh and rectus abdominis myocutaneous flap. The postoperative course was uneventful. The patient subsequently developed brain metastasis and died within 52 months after sternal resection. The stability of the chest wall was well preserved for the duration of follow-up. PMID- 15828260 TI - [The descriptions on disorders of urination in the Hippocratic Collection]. AB - PURPOSE: Hippocrates was a famous physician of Kos in ancient Greece (c. 460 B.C.). His works were later described in the Hippocratic Collection, in which I studied disorders of urination. MATERIALS AND METHODS: I collected mentions of disorders of urination in the Hippocratic Collection (Roeb, Otsuki and Kon editions) to compared description of disorders of urination between the Kos and Knidos Schools. RESULTS: Disorders of urination were mentioned in 67 parts (sentences or sections): difficult urination in 50, retention of urine in 15 and urinary incontinence in two or three (presumably complicated by difficult urination in one). sigmastrangourih (strangourih) and deltadusourih (dusourih) were counted in 20 and 30 parts, respectively. sigmastrangourih was used in 12 (60%), 5 (25%) and 3 (15%) respective descriptions by Kos, Knidos, and unspecified School, while deltadusourih was used in 13 (43%), 17 (57%), and no descriptions; the correlation was not significant, (chi-squared test, p > 0.05). Descriptions of "stoppage of the urine or bladder", "blockage of the urine", "inability to pass urine", and "check" in classic Greek may mean retention of urine, which also is suspected in the case of severe deltadusourih and sigmastrangourih. Retention of urine only appeared in descriptions by the Kos School, except for one of origin not specified between the Kos and Knidos Schools. The terms "deltadusourih" and "sigmastrangourih" are the origins of the English terms "dysuria" and "strangury". Dysuria ("dys + urine + ia") denotes painful or difficult urination, while strangury ("drop + urine") denotes slow and painful discharge of urine. Deltadusourih and sigmastrangourih with severe pain appear in descriptions by the Kos School. However, in classic Greek, these words by no means always meant painful urination, which differed partially from present day meanings of dysuria and strangury. There were deltadusourih, sigmastrangourih, and retention of urine accompanied by severe, slight or no pain. The former degree of pain may have included urinary tract infection and/or prostatitis, with the latter, including prolape of uterus or bladder. Sigmastrangourih was a chronic disease of a nonfatal natures, unless complications occurred. "Unconsciously passed a copious discharge of urine" may refer to urinary incontinence, which was seen in one description by Kos and two by Knidos. Two types of urinary incontinence were described, copious and drop urine wise urinary incontinence; the former may represent neurogenic bladder caused by brain dysfunctions, while the latter may represent overflow incontinence. Urethral catheterization and diuretics already were mentioned in the Hippocratic Collection. Bleeding and anodyne were used in treatment of deltadusourih and sigmastrangourih. CONCLUSIONS: Disorders of urination appeared in four categories, namely difficult urination or dysuria: dropwise urine or strangury; urinary retention : and urinary incontinence. Severe disorders of urination appear frequently in descriptions by the Kos School. PMID- 15828261 TI - [Usefulness of extension of disease as a prognostic factor in relapsed prostate cancer patients of stage D]. AB - OBJECTIVE: The prognosis of prostate cancer has been evaluated by clinical stage or pathological grade. PSA parameters including PSA density and PSA doubling time have not always precisely reflected the prognosis of prostate cancer. The aim of this study was to evaluate PSA parameters and extension of disease (EOD) grade as prognostic factors for relapsed prostate cancer. METHODS: The relationship between PSA parameters or EOD grade, and survival of 29 stage D patients with relapsed prostate cancer after initial hormone therapy was examined. RESULTS: Only EOD grade was an independent prognostic factor, even for cause-specific survival period and survival period after relapse. CONCLUSION: EOD grade was a significant prognostic factor, and in particular, very useful as a prognostic factor for patients with bone metastasis. PSA value was not always associated with tumor volume, and therefore it is not an independent prognostic factor. PMID- 15828262 TI - [Safety and efficacy of a new vessel-sealing device for laparoscopic varicocelectomy]. AB - PURPOSE: To evaluate the safety and efficacy of a new vessel-sealing device (LigaSure) for laparoscopic varicocelectomy. MATERIALS AND METHODS: To evaluate the safty, after elective varicocelectomy, the internal spermatic veins (n = 8) were sealed ex vivo with a 5-mm laparoscopic Maryland-style LigaSure (LigaSure Lap), and bursting pressures were measured. To evaluate the efficacy, a retrospective review of our clinical experience with LigaSure (n = 13) and clip (n = 13) in laparoscopic bilatelal varicocelectomy from June 2000 to October 2002 was performed and certain parameters were abstracted, including operative time, estimated blood loss and perioperative complications. RESULT: In the ex vivo study the mean varicocele vessele diameter was 2.5 +/- 0.8 mm (mean +/- SD) and bursting pressure was 449 +/- 95.2 mmHg. Six were burst in normal vessel walls at the average bursting pressure of 442 mmHg and two in the occlusions at the average bursting pressure of 508 mmHg. The difference was not statistically significant. Reliable vessel sealing was achieved in all the patients. The mean operative time showed significant decrease in the LigaSure group compared with the clip group, 70 +/- 20 minutes versus 117 +/- 27 minutes (p < 0.05). Estimated blood loss was minimal and no perioperative complications occurred in both groups. CONCLUSIONS: Our study suggests that a vessel-sealing device appears to be a safe and feasible alternative to the clip in laparoscopic varicocelectomy. It offers the advantage of reducing operative time. PMID- 15828263 TI - [Analysis of mailed questionnaire for female sexual dysfunction after intra pelvic surgery]. AB - OBJECTIVE: Female sexual dysfunction is an unfavorable complication for women after intra-pelvic surgery. However, there has not been enough data obtained to analyze the inconvenience to female patients of intra-pelvic surgery. To clarify the categories of female sexual dysfunction after intra-pelvic surgery, we analyzed mailed questionnaires obtained from female patients who had undergone mono-therapy of intra-pelvic surgery. MATERIALS AND METHODS: A mailed questionnaire regarding female sexual dysfunction was sent to 174 patients, including 118 that had undergone a hysterectomy (Group A) and 56 ostomates (Group B) and the results were analyzed. These patients had received only intra-pelvic surgery, without radiation or chemotherapy. Of the 56 patients in Group B, 50 with rectal cancer had received a colostomy, and 6 with bladder cancer had received ileal conduits. The content of the questionnaire was as follows: age; with or without a male partner; key person for consultation in regard to sexual dysfunction; importance of sex life. Also, a before and after operation comparison was made regarding sexual frequency, sexual desire, genital response, genital transformation, dyspareunia, genital pain and itching, and orgasm. RESULTS: Seventy-eight of 174 (45%) patients returned the questionnaire. Nineteen patients without a male partner and 18 patients with a male partner had no sexual intercourse, very low sexual desire, and did not consider their sex life of importance. Twenty-five patients with a male partner (18 in Group A and 7 in Group B) had no trouble in their sexual activity after the operations. However, the other 16 patients (5 in Group A and 11 in Group B) had unsatisfactory sexual intercourse after their operations because of a decrease in sexual desire, poor genital response, and/or decrease of orgasms. Furthermore, the patients in Group B suffered significantly from sexual trouble compared with the patients in Group A. Especially, all the patients in Group B complained that their stoma prevented satisfactory sexual intercourse. CONCLUSIONS: Based on this data, we do not need to worry about female sexual dysfunction for patients without a male partner and patients with a partner but who are not sexually active before their operations. More than 60% of patients who had been sexually active before their operations maintained fair sexual activity after intra-pelvic surgery. However, we have to take care of the approximately 40% of patients who suffer from female sexual dysfunction after their operation, especially the ostomate patients. PMID- 15828264 TI - [A case of prostatic cystadenoma]. AB - We report a case of prostatic cystadenoma. A 81-year-old man visited our hospital with the chief complaint of urinary difficulty. RUG and DIP showed bladder and prostatic urethra deviated to the left side. TRUS, CT and MRI revealed multilocular cyst in the pelvic cavity. Resection of the cystic wall was performed under the diagnosis of pelvic cyst. The cysts were originated from the right side of the prostate and adhered with the bladder severely. The histopathological diagnosis was prostatic cystadenoma. Fortyfive cases of prostatic cyst in the Japanese literature are reviewed. PMID- 15828265 TI - [Two cases of second bladder cancer after radiotherapy for prostate cancer]. AB - Second cancer after radiotherapy is defined that more than five years have passed from radiotherapy for primary cancer in case of solid and different histological cancer and second cancer exists into or near radiotherapy area. While it has been not frequent that a treatment for urological cancers causes second cancer, there is the possibility that second cancer increases by recent increase of radiotherapy for prostate cancer. We report two cases of second bladder cancer after radiotherapy for prostate cancer. It is important to take second cancer into consideration for long-term after the patients are treated with radiotherapy. PMID- 15828266 TI - [Omega-3 fatty acids in mood disorders]. AB - The etiology and treatment of mood disorders has not yet been elucidated. Omega (omega)-3 fatty acids are essential fatty acids, which cannot be synthesized in the human body. Eicosapentanoic acid (EPA) and docosahexaenoic acid (DHA) are representative omega-3 fatty acids which are found in fish (eg., mackerel, herring, Chinook salmon) and vegetables (eg., flax, walnut, canola). The peripheral level of EPA and DHA decrease in patients with major depression, and EPA is useful for its treatment. Further research is required on omega-3 fatty acids in patients with mood disorders. PMID- 15828267 TI - [Experiences in the United States--a difference in the status in the United States and that in Japan]. AB - Child and adolescent psychiatry has been established as a subspecialty in the United States. The systematic education, training and research opportunities are currently available and child and adolescent psychiatry is a major part of psychiatric practice in the US. However, there are many burdens to overcome to provide adequate child mental health care. Several important focus on child mental health care in US are to 1) improve the assessment of and recognition of mental health needs in children, 2) promote scientifically-proven and evidence based interventions and 3) increase access to and coordination of quality mental health care services. To understand the current status and problems in child and adolescent psychiatry in US may help the Japanese psychiatrists to develop and establish child and adolescent psychiatry in Japan. PMID- 15828268 TI - [Psychiatric therapy for adolescent patients]. AB - Most children who go through adolescence manage to overcome the "storm of adolescence," but some children are not so fortunate and have difficulty in coping. They may feel that their inability to cope is unique or that they will forever be handicapped or diseased as a result. In reality, the trials of adolescence, combined with a sub optimal personal life often result in setbacks. Adolescence, by definition is a time of development. And this development is extensive; physical, mental, sexual and self-awareness. Personality change is inevitable. Over a relatively short period, young children evolve into adults. Every one experiences these drastic changes, but to each child, it seems that he/she is going through the experience alone, for the first time. We must convey to these children that this situation is normal, and is inevitably stressful. If we can succeed in even a small way to do this, I believe we can find a way to help adolescent children. PMID- 15828269 TI - [Current topics in vitreoretinal diseases]. PMID- 15828270 TI - [Bibliometric analysis of the current international ophthalmic publications]. AB - PURPOSE: To assess the current status of international ophthalmic publications. METHODS: A collection of 55,591 original articles were found by an on-line National Library of Medicine database Medline search for 32 international ophthalmic journals during a 15-year period from 1988 to 2002 (internet access, November 11-13, 2003). RESULTS: The contributions to international ophthalmic publications were by 49.5% from North America, 31.3% from Western Europe, 15.1% from Asia and Oceania, 2.2% from Middle East, 0.85% from Central and South America, 0.53% from Eastern Europe, and 0.47% from Africa. Countries of Asia and Oceania showed an increasing trend in contributions while North America had a decreasing productivity in a relative sense. The top 10 productive countries were USA, United Kingdom, Japan, Germany, Canada, Australia, Italy, Netherlands, Sweden, and France. Among the Asian countries India ranked 13th, China 18th, and Korea 21st. When related to population, small countries such as Israel, France, Finland, Sweden, and Denmark were more productive. When related to economic productivity as defined by GDP, Israel, the United Kingdom, Australia, Finland, and Sweden were among the most productive countries, whereas rich countries such as Japan and Germany had a lower number of publications relative to their GDP. As regards clinical research in terms of randomized controlled trials, The USA was by far the most productive. The number of authors per article has shown an increasing trend worldwide, so that Japan and France had a significantly larger proportion of multiauthored articles. CONCLUSIONS: There is an increasing trend in the productivity of international ophthalmic publications from non-English speaking countries including Japan, China, and Korea. PMID- 15828271 TI - [Opacification of Hydroview intraocular lenses necessitating lens replacement]. AB - PURPOSE: We report the opacification of Hydroview intraocular lenes necessitating lens replacement. SUBJECTS AND METHODS: This study includes 9 eyes of 7 patients (4 eyes of 3 men and 5 eyes of 4 womens, 52-92 years old) who underwent lens replacement at our hospital. A clinicopathological study and speculation on the incidence of Hydroview opacification were carried over. RESULTS: All patients had hypertension and 3 had diabetes mellitus. Four eyes of 3 cases and 2 eyes of 2 cases demonstrated diabetic retinopathy and pseudoexfoliaton syndrome, respectively. Four of 6 cases, who underwent bilateral lens implantation, showed unilateral opacification. The duration from lens implantation to exchange ranged from 17 to 38 months. Slit-lamp examination revealed generalized opacification with white granular deposits on the lens optic surface, but transparency inside the optic. Three explanted lenses were stained with 1% alizarin red, and were positive for calcium in all cases. The remaining 6 lenses were analyzed with scanning electron microscopy, and showed electrondense granular deposits of 1 to 4 microm diameter. It is theorized that the incidence of Hydroview opacification and necessity of replacement were more than 3.8% and 3.6%, respectively, from the data at one hospital. CONCLUSION: Calcium deposition on Hydroview might occur in more than 3.8% of cases. PMID- 15828272 TI - [Vitrectomy for proliferative diabetic retinopathy]. AB - PURPOSE: We reviewed the outcome of vitrectomy for proliferative diabetic retinopathy (DR) and evaluated factors affecting the final visual outcome. METHODS: We performed primary vitreous surgery for proliferative DR in 148 eyes of 118 cases in three years from July 1999 to August 2002. All cases were followed for at least 3 months. We excluded vitreous surgery for diabetic maculopathy. Ages ranged from 24 to 80 (mean 57) years. Average postoperative follow-up period was 15 months. We evaluated the stage of DR by the new Fukuda classification. RESULT: Preoperative classification consisted of BIV (54 eyes, 36%), BV (94 eyes, 64%), and BV + VI (36 eyes). Final visual acuity was improved by 2 lines or more in 102 eyes (69%), remained unchanged in 28 eyes (19%), and decreased by two lines or more in 18 eyes (12%). There was a statistical correlation between preoperative visual acuity and final visual acuity. Earlier stages of DR had better visual outcome. Compared to the surgical outcome in the 1990s, the percentage of worsened eyes decreased. CONCLUSION: Vitrectomy for proliferative DR may be beneficial if performed in the earlier stages of DR or if the patient has better visual acuity before vitrectomy. PMID- 15828273 TI - [Evaluation of macula in patients with branch retinal vein occlusion using multifocal electroretinogram and optical coherence tomography]. AB - PURPOSE: To evaluate the functions of the macular area in patients with branch retinal vein occlusion (BRVO) by using multifocal electroretinograms (mERG) and optical coherence tomography(OCT). METHODS: mERGs were recorded from 18 eyes of 18 patients with BRVO. An array of 103 hexagonal elements was displayed on a monitor. The latency (ms) and response density (nV/deg2) of mERGs were measured for 7 central locations. The peak and troughs were labeled N1, P1 and N2, respectively. OCT was used to measure the foveal retinal thickness. RESULTS: Statistically significant correlation was found between visual acuity(log MAR) and P1-N1 response density (r = - 0.629, p < 0.05) and N2 latency(r = 0.619, p < 0.05). Foveal retinal thickness had the significant statistic correlations with P1-N1 response density (r = -0.750, p < 0.0001), P1-N2 response density (r = 0.520, p < 0.05) and N2 latency (r = 0.488, p < 0.05). CONCLUSIONS: In 18 BRVO patients, mERGs from the central retinal area were significantly correlated with foveal retinal thickness measured by OCT and with visual acuity. The mERG recording is sensitive to morphological changes and functional disorders induced by BRVO. PMID- 15828274 TI - [Effectiveness of plasma exchange in Miller Fisher syndrome--a case report]. AB - BACKGROUND: We report a successful treatment by plasma exchange(PE) in the case of Miller Fisher syndrome. CASE: A 71-year-old woman rapidly developed diplopia and unsteady gait, and was admitted to Minami-matsuyama Hospital on day 3 of her illness. Ophthalmological and neurological examination on admission revealed bilateral complete external ophthalmoplegia with moderate mydriasis, cerebellar ataxia, and weakness of biceps reflexs, but her consciousness was intact. Protein concentration in the cerebrospinal-fluid was slightly increased and serum anti GQlb IgG antibody was positive. With the diagnosis of Miller Fisher syndrome, PE was carried out on days 12, 14, and 16. After PE, ophthalmoplegia and cerebellar ataxia improved markedly. The ocular symptoms resolved after a month. CONCLUSION: The findings indicate that PE is a beneficial treatment in Miller Fisher syndrome during the acute phase of the disease. PMID- 15828275 TI - [Roles and activation mechanisms of Nox family oxidases in animals]. PMID- 15828276 TI - [Role of pathogen-responsive oxidative burst in plants]. PMID- 15828277 TI - [Oxidative damage, maintenance, and higher structure of mitochondrial DNA]. PMID- 15828278 TI - [Biological consequences and base excision repair mechanisms of oxidative base damage in DNA]. PMID- 15828279 TI - [Molecular mechanisms in aberrant splicing in neurological disorders]. PMID- 15828280 TI - [Vacuolar processing enzyme exhibiting caspase 1-like activity is involved in plant programmed cell death]. PMID- 15828281 TI - [Proteoglycans inducing plant cell differentiation]. PMID- 15828282 TI - [Molecular mechanism of protein import through receptor complexes: implication of the complex structure of outer membrane receptor and colicin E3]. PMID- 15828283 TI - [Laminin active peptides as a powerful tool for biomedical materials]. PMID- 15828284 TI - [Superselective intraarterial chemotherapy using low dose CBDCA and Pirarubicin with concurrent radiotherapy for head and neck cancer]. AB - To study the effects of superselective intraarterial chemotherapy with low-dose CBDCA, Pirarubicin, and concurrent radiotherapy on head and neck cancer, we compared primary cancer response and histopathological effective grades in 66 patients (more than T2) divided into radical and preoperative radiotherapy groups. The radical group (n=33) showed a 75.7% response in primary cancer, i.e. 54.5% complete remission and 21.2% partial remission. The preoperative group (n = 33) showed 39.4% complete remission when the histopathological effective grade was higher, and 57.6% partial remission when the grade was lower. Cancer response was better in the oral cavity, mesopharynx, and hypopharynx than in the parasinus. In the preoperative group, 5-year overall survival was 84.4% when the effective grade was higher, and 29.4% when the grade was lower. Survival differed significantly (P<0.01) between higher and lower grades. Additional postoperative therapy is thus essential in patients with lower grades of histopathological effectiveness. PMID- 15828285 TI - [Horizontal canal benign paroxysmal positional vertigo--its affected side and horizontal semicircular canal function]. AB - We report horizontal canal BPPV (HC-BPPV) targeting its pathophysiology, the affected side, and the function of the horizontal semicircular canal, together with a review of the literature. Subjects were 13 patients with HC-BPPV visiting our vertigo outpatient clinic at Nara medical university hospital and a related hospital in the 2.5 years from January 2000 to June 2002. Subjects were classified into 7 with canalolithiasis and 6 patients of cupulolithiasis after a neurotological examination. CP was positive in 54% of all patients, 71% of those with canalolithiasis, and 33% of those with cupulolithiasis. To determine the affected side in HC-BPPV, we used the affected side by using the law of Ewald in canalolithaisis patients and the detection of a neutral diminishing nystagmus in cupulolithiasis patients. CP positive in caloric testing indicated insignificant dysfunction of the horizontal semicircular canal in canalolithiasis patients compared to that in cupulolithasis patients. The mechanism behind caloric nystagmus was thought to be a convection of endolymphatic fluid interrupted consequently by an otolith in the semicircular canal in canalolithiasis patients. In contrast, CP was positive in cupulolithiasis patients regarded as having no convection of endolymphatic fluid. The mechanism causing a difference in caloric test results between canalolithiasis and cupulolithiasis patients thus requires a larger patient population and further examination to be conclusive. PMID- 15828286 TI - [Promoter hypermethylation of tumor-associated genes in head and neck cancer]. AB - An important pathway of gene transcriptional inactivation is hypermethylation at the CpG islands of promoter regions. Some tumor suppressor genes have been reported to harbor promoter hypermethylation in head and neck cancer. We studied DNA hypermethylation of 4 genes in 42 cases of primary head and neck cancer. We applied methylation specific PCR for p16, RAR-beta, RASSF1A, and Fhit genes. Hypermethylation was detected at p16 in 43%, at RAR-beta in 40%, at RASSF1A in 12%, and at Fhit in none of the cases. Hypermethylation of at least one gene was detected in 26 (62%) of the 42 cases. No significant correlation was seen between methylation status and clinicopathological findings or prognosis. Hypermethylation of several tumor-associated genes plays an important role in tumorigenesis of head and neck cancer. We discuss clinical implications and their application. PMID- 15828287 TI - [Bilaterality in acute low-tone sensorineural hearing loss]. AB - Bilaterality in acute low-tone sensorineural hearing loss (ALHL) is more generally recognized than that in idiopathic sudden sensorineural hearing loss. Subjects were 274 patients diagnosed with ALHL based on criteria of a study group of the Ministry of Health, Labor and Welfare of Japan, i.e., total of 3 low tone hearing of 70dB or more and, a total of 3 high-tone hearing of 60dB or less, and treated at the departments of otolaryngology at Yamanashi University and Suwa Central Hospital. ALHL involving bilateral ears symptoms and/or bilateral hearing impairment conforming to diagnostic criteria was selected and summarized. Clinical ear symptoms, clinical test results, and hearing levels (total 3 low tone hearing, 1kHz, and total of 3 high-tone hearing) were statistically analyzed. We also reviewed Japanese clinical reports of ALHL that include bilateral cases. In 32 cases (11.7%) of 274 cases, both ear symptoms and hearing impairment were bilateral. In 22 (8.0%) of the 274, bilateral ear symptoms were present, but showed unilateral hearing loss conforming to diagnostic criteria. Another 22 (8.0%) out the 274 reported unilateral ear symptoms, but hearing tests indicated bilateral ALHL. A total of 76 cases (27.7%) of the 274 had bilaterality in either ear symptoms or hearing loss. Our review indicated that 9.0% (162 of 1803) ALHL patients were bilaterally affected, possibly indicating that AIHL includes a larger number of bilateral cases than currently assumed, if the opposite side were given a especially detailed clinical interview. Statistical analysis (Mann Whitney test, P<0.01) of hearing of unilateral cases indicated that 3 low tone hearing was more affected than 3 high-tone hearing, even on the normal side. These results indicate that ALHL tends to be bilateral, possibly due to the mechanism of pathogenesis, and also that the mechanism may include both local and general conditions. This more closely resembles Meniere's disease than idiopathic sudden sensorineural hearing loss. Both sides of bilateral cases were not usually the same in hearing patterns, glycerol test results, or prognosis. A statistically significant difference (Mann Whitney test, P<0.01) in total of 3 low tone hearing was seen between worse and better sides in bilateral cases. The degree of disease on both sides in bilateral cases thus was not always the same. Bilateral cases may result from both the influence of general conditions such as fatigue, stress, and lack of sleep and local conditions such as pathogenesis of endolymphatic hydrops that may cause differences in both ears. No clear difference was seen in clinical symptoms, hearing levels, and clinical examination, e.g., Schellong and glyceol tests, between unilateral and bilateral cases. Bilateral cases had a poorer prognosis (lower complete recovery ratio; chi2 test P<0.01) than unilateral cases. Our results indicated that cautious evaluation of opposite ear is necessary in diagnosis, treatment, and prognostication of ALHL. PMID- 15828288 TI - [Two cases of Meniere's disease with drop attacks]. AB - Drop attacks are sudden falls without concurrent vertigo whose etiology may be unknown. Drop attacks are also associated with cardiac, cerebrovascular, psychogenic, and vestibular disorders, in addition to seizures. Vestibular-based drop attacks without loss of consciousness can occur in patients with Meniere's disease. We present 2 cases of drop attack in patients with Meniere's disease. Case 1, a 65-year-old man, experienced 4 such attacks and case 2, a 55-year-old woman, experienced 20 within 2 years of Meniere's disease onset. Case 1 enjoyed spontaneous remission. In case 2, selective serotonin reuptake inhibitor (SSRI) administration suppressed attack frequency. Anxiety may predispose individuals to drop attack. Etiologically, inadequate stimulation of otolith organs may induce a sudden vestibulo spinal reflex that, in turn, causes sudden falls. To correctly diagnose drop attacks, the patient must exhibit sudden falls, and transient ischemic attack should be carefully ruled out. PMID- 15828289 TI - [Toxic shock-like syndrome of the neck, a case report]. AB - Streptococcal toxic shock-like syndrome (TSLS) is known to progress rapidly into septic shock and have a high mortality of 40%. We report a rare case of neck infectious disease caused by Group A hemolytic streptococcus pyogenes. A 65-year old man, hospitalized for eyelid swelling and dyspnea suffered shock, dyspnea, and pressure reduction on the day he was hospitalized and died of multiple organ failure 18 days later. Despite treatment with antibiotics, tracheostomy, immunoglobulin, etc. We suspected that he had been infected via a laceration of the external auditory meatus. Accurate diagnosis followed by prompt, radical treatment is essential if TSLS is to be treated successfully. PMID- 15828290 TI - Coronary artery surgery in Japan. PMID- 15828291 TI - Off-pump coronary artery bypass grafting attenuates proinflammatory markers. AB - OBJECTIVE: Cardiac surgery with cardiopulmonary bypass (CPB) has been considered the main causative factors of postoperative inflammatory reactions. The aim of this study was to compare surrogate markers of the proinflammatory response in patients who underwent coronary artery bypass grafting (CABG) with or without CPB. METHODS AND RESULTS: Twenty patients undergoing first-time CABG were enrolled in the study, 10 with and 10 without CPB. Blood samples were drawn at the following times: at the anesthetic induction, the end of surgery, and thereafter at 12 and 24 hours postoperatively. Neutrophil elastase, interleukin (IL)-6 , and serum soluble Fas were chosen to evaluate the extent of the systemic inflammatory response. The groups were similar in terms of age, gender ratio, number of grafts per patient. There were no operative mortality or serious postoperative complications. Two of each group received blood transfusion postoperatively. Neutrophil elastase showed a significantly higher value in the on-pump group compared with the off-pump group at the end of surgery. Soluble Fas level showed a higher value at the end of surgery compared with baseline, while it had no significant changes in the off-pump patients. IL-6 levels in the on pump group were consistently higher compared to the off-pump group but showed no statistically significant differences between the groups. CONCLUSION: Compared with off-pump CABG, on pump CABG induced higher serum levels of proinflammatory markers including neutrophil elastase and soluble Fas. PMID- 15828292 TI - Simultaneous operation of off pump coronary artery bypass and abdominal aortic aneurysm repair. AB - OBJECTIVES: Coronary artery disease (CAD) and abdominal aortic aneurysms (AAA) commonly coexist. However, each disease treatment complicates the management of the other. In this study, we evaluate whether a simultaneous operation of AAA repair and off pump coronary artery bypass (OPCAB) would be safe and acceptable, compared with either procedure alone. SUBJECTS AND METHODS: We retrospectively reviewed all patients who underwent simultaneous AAA repair and OPCAB (AAA/OPCAB, n=18), compared AAA repair alone (AAA, n=239) and OPCAB alone (OPCAB, n=137) from June 1999 to December 2003. There were no significant differences with regard to age or gender, but the AAA/OPCAB group had significantly larger aneurysms (60.6 vs. 53.2 mm) and significantly lower ejection fractions (EF) (54.9 vs. 60.3%). RESULTS: The patients in the AAA/OPCAB group underwent a significantly longer operative time than AAA, OPCAB (403 vs. 360, 296 minutes, respectively), there was significantly greater blood loss (726 vs. 426, 462 ml), and more transfusion required (8.13 vs. 1.69, 2.8 units). The number of bypass grafts in AAA/OPCAB group (1-5 per patients) was significantly smaller (1.78 vs. 2.93). The AAA/OPCAB patients had a significantly longer hospital stay than the AAA (38 vs. 22 days), but was not significantly longer than the OPCAB. There were no significant differences with regard to the morbidity and mortality rate among the three groups. CONCLUSION: This study suggests that the simultaneous operation of AAA and OPCAB can be done with the same morbidity and mortality as independent surgical procedures. PMID- 15828293 TI - Characteristic changes in cerebral perfusion during on-pump and off-pump coronary artery surgery. AB - OBJECTIVE: We investigated the cerebral perfusion status during on-pump or off pump coronary artery bypass grafting (CABG). METHODS: We monitored somatosensory evoked potential (SEP) and regional cerebral oxygen saturation (rSO2) as parameters of cerebral perfusion in an on-pump group (n=10) and an off-pump group (n=16). The percent changes from control values were calculated before, during, and after aortic clamping, and after weaning from cardiopulmonary bypass, in the on-pump group. In the off-pump group, these were calculated before, during, and after heart displacement for distal anastomosis. RESULTS: In the on-pump group, the amplitudes of the SEP were significantly enhanced during and after aortic cross-clamping and were associated with a significant decrease in rSO2. Latency was prolonged immediately after aortic cross-clamping, but was shortened afterwards. There was little change in these parameters throughout the operation, in the off-pump group. CONCLUSIONS: Cerebral perfusion remains stable during off pump CABG. The etiology of a simultaneous increase in SEP amplitude and decrease in rSO2 during the rewarming period in the on-pump group requires further investigation. PMID- 15828294 TI - Papillary fibroelastoma of the mitral valve associated with rheumatic mitral valve stenosis. AB - With the advent of echocardiography, diagnosis of papillary fibroelastoma in living patients has been made possible, yet papillary fibroelastoma found in the living remains a very rare cardiac tumor. We report a case of papillary fibroelastoma of the mitral valve with rheumatic mitral valve stenosis. A 68-year old woman was referred to our hospital with a mitral valve tumor and rheumatic mitral valve stenosis. She underwent anticoagulation therapy with Warfarin for 8 years since having a cerebral embolization. Echocardiography revealed a mass attached to the mitral valve, with severe mitral valve stenosis. Electrocardiography demonstrated a chronic atrial fibrillation. Tumor excision with mitral valve replacement and maze procedure were performed. Both the surgical and histological findings depicted papillary fibroelastoma. The postoperative course was uneventful and the patient has remained symptom-free one year after surgery. PMID- 15828295 TI - Left ventricular true aneurysm with pseudoaneurysm detected five years and nine months following repair for oozing type free wall rupture. AB - A case of true aneurysm of the left ventricle associated with pseudoaneurysm was treated surgically. The condition was detected five years and nine months following repair of an oozing type left ventricular free wall rupture due to myocardial infarction. Over this period, chest radiographs showed gradual cardiomegaly with prominence of the left fourth arch. PMID- 15828296 TI - Postoperative prone position management of tetralogy of fallot with absent pulmonary valve syndrome. AB - Tetralogy of Fallot with absent pulmonary valve syndrome is commonly associated with respiratory failure both before and after surgery. This report describes our experience using prone positioning with bilateral pillows to avoid compression of the anterior chest wall after surgery. In the case here, the patient's respiratory distress was improved by this positioning. Prone position and avoiding anterior chest compression has an effect on severe respiratory distress of tetralogy of Fallot with absent pulmonary valve syndrome. PMID- 15828297 TI - Intrapulmonary schwannoma. AB - A 58-year-old woman was admitted due to an abnormal shadow on chest X-ray, without any symptoms. Chest computed tomography showed a round mass in the anterior segment of the right upper lobe. Segmentectomy was performed and histopathological examination revealed a primary neurogenic tumor of Schwann cell origin. Immunohistochemical staining demonstrated the presence of S-100 protein in the tumor cells. We present a case of intrapulmonary schwannoma and review 62 cases of primary schwannoma of the lung. PMID- 15828298 TI - Bilateral multiple pulmonary sclerosing hemangioma. AB - We present herein a rare case of bilateral pulmonary sclerosing hemangioma, for which a differential diagnosis was made from metastatic lung tumors. A 32-year old asymptomatic woman was referred to our hospital for further evaluation of abnormal chest shadows. A chest computed tomogram revealed two round, well circumscribed masses in both sides of the lungs. Metastatic lung tumors were suspected, however, a primary lesion was not detected by several examinations. Thus, simultaneous video-assisted thoracic surgery for the bilateral tumors was performed. The tumors, measuring 16x13x12 mm in the left lung and 27x24x20 mm in the right lung, were resected, and then pathological examination confirmed the diagnosis of sclerosing hemangioma. Her postoperative course was uneventful and she has been doing well without any sign of recurrence. PMID- 15828299 TI - Thoraco-pleuropneumonectomy for rupture of a huge chronic hematoma of the thorax. AB - A 67-year-old woman with a huge chronic expanding hematoma of the thorax due to previous tuberculous pleuritis was referred to our hospital with frequent hemoptysis. The hematoma had ruptured into the lung parenchyma. The patient had undergone apical thoracoplasty 40 years earlier. To provide complete resection of the huge cavity in the costodiaphragamatic recess, thoraco-pleuropneumonectomy was performed, and the patient was able to resume daily activities a few months after the operation. PMID- 15828300 TI - Reoperation for a Starr-Edwards aortic prosthetic valve 33 years after initial implantation. AB - A 40-year-old woman admitted with heart failure had undergone aortic valve replacement with a Model 2310 Starr-Edwards valve due to aortic regurgitation 33 years previously. She had been followed up for several years, but discontinued follow-up and medication (including Warfarin) for the past 25 years. Echocardiography demonstrated marked dilatation and thickening of the left ventricle, and the peak pressure gradient of the prosthesis was measured as 87.9 mmHg. Under the diagnosis of chronic aortic valve prosthesis-patient mismatch with subsequent severe left ventricular dysfunction, the Starr-Edwards valve was explanted and replaced with a 23 mm St. Jude Medical prosthetic valve. The removed valve showed minimal cloth wear except for a small part of the strut. The postoperative echocardiography demonstrated recovering of left ventricular function. To our best knowledge, this case presents the longest duration for a surgically explanted Starr-Edwards aortic prosthetic valve in Japan. PMID- 15828301 TI - Successful surgical treatment for diffuse esophageal spasm. AB - A 74-year-old man, who had previously received curative distal gastrectomy for gastric cancer, was admitted to our hospital with severe dysphagia and weight loss. Barium swallow examination revealed the esophagus to have the corkscrew appearance characteristic of diffuse esophageal spasm (DES). This diagnosis was confirmed by esophageal manometry, which revealed intermittent, simultaneous, high-amplitude (30-100 mmHg) contractions after 65% of wet swallows. The muscle layer was also found to be thickened throughout the spastic region. Long esophagomyotomy with fundoplication was performed after treatment with medication proved ineffective. Myotomy proceeded superiorly to the area under aortic arch and inferiorly 3 cm into the cardiac portion. Fluoroscopy of the esophagus after the operation showed the spastic changes to be absent, and the patient showed improved clinical signs. We therefore recommend long myotomy of the esophageal wall with antireflux surgery for DES with sever dysphagia that is resistant to conservative treatment. PMID- 15828302 TI - Coccidioidomycosis of the lung contracted abroad. AB - Coccidioides immitis is endemic in the southwestern United States. Coccidioidomycosis is rare in Japan, but the number of coccidioidomycosis cases from overseas is expected to increase markedly in the near future. A 33-year-old Japanese man with coccidioidomycosis who had lived in Arizona presented to our hospital. We performed wedge resection of the lung by video-assisted thoracoscopic surgery (VATS), and were able to completely remove the lesion. VATS wedge resection of the lung is effective in treating localized coccidioidomycosis. Before the operation, we could not confirm the existence of satellite lesions, but microscopically the main lesion was accompanied by several daughter lesions. It is very important that the surgical margin be sufficient to achieve complete resection. PMID- 15828303 TI - Performance of the Fontan procedure using extra-cardiac direct anastomosis on patients treated for penicillin-resistant Streptococcus pneumoniae endocarditis. AB - Streptococcus pneumoniae is now a rare cause of endocarditis in humans. We report a patient with a double outlet right ventricle and mitral atresia, who underwent the Fontan procedure without prosthetic materials after treatment for penicillin resistant Streptococcus pneumoniae endocarditis. Postoperative infectious endocarditis was not found. In patients with a history of infectious endocarditis, direct anastomosis of the main pulmonary artery and inferior vena cava would reduce the risk of recurrent infectious endocarditis. PMID- 15828304 TI - [New insights into the pathology of nasal polyposis: the role of superantigens and IgE]. AB - Although the etiology of nasal polyposis is still not revealed, insights in the pathogenesis have largely expanded over the last years. Usually nasal polyps occur in adults, are bilateral and are characterized by a manifest tissue eosinophilia. Deposition of plasma-proteins (albumin), potentially driven by subepithelial eosinophilic inflammation, seems to be an early and key pathogenic factor in the development of nasal polyps. Accumulation and activation of eosinophils is favoured by low TGF-beta1 concentrations and overproduction of IL 5 and eotaxin. In nasal polyps high IgE concentrations are measured. Our findings indicate that this IgE is produced locally. Total IgE and the expression of specific IgE is unrelated to skin prick tests, but correlates with the degree of eosinophilia. In addition, we demonstrated the organisation of secondary lymphoid tissue in nasal polyps and a polyclonal hyper-immunoglobulinemia E, associated with the presence of IgE specific to Staph. aureus enterotoxins (SAE), colonization with Staph. aureus and increased eosinophilic inflammation in a relevant subgroup of NP patients (about 50%). In about half of the nasal polyps we thus find a local immune response against SAE. SAE can hereby act as conventional allergens, triggering T- en B-cells to produce sIgE against SAE. On the other hand, SAE can also act as superantigens and induce polyclonal B-cell activation and hyper-immunoglobulinemia. In addition, the presence of IgE antibodies to SAEs seems to be associated with the severity of asthma and nasal polyposis disease. Nasal and oral corticosteroids are currently the standard treatment for NP. This treatment however, is not always sufficient and oral corticosteroids have several side effects. Often surgery is required, which in turn is not free of complications and recurrencies. Increasing insights in the pathophysiology of NP opens perspectives for new pharmacological treatment options, with eosinophilic inflammation, IgE and Staph. aureus as potential targets. PMID- 15828305 TI - [Detection of minimal residual disease in children with neuroblastoma]. AB - The presence of residual cells in bone marrow specimens of stage 4 patients, is an important prognostic parameter in neuroblastoma. Sensitive and specific standardized techniques are a prerequisite to detect and quantify these residual cells. In this respect, we evaluated an immunocytochemical test, a multiparameter flow cytometric assay and quantitative real-time RT-PCR. The latter technique was the most sensitive one in our hands. Although it is a simple and highly reproducible test, RT-PCR does not allow visualisation of neuroblastoma cells. On the contrary, the immunocytochemical APAAP assay is slightly less sensitive but can discriminate neuroblastoma cells morphologically. The combination of both assays seems to be ideal for monitoring residual cells in bone marrow of neuroblastoma patients. The flow cytometric multiparameter assay particularly offers additional information in GD2-negative neuroblastoma. Preliminary results of our study in children with stage 4 neuroblastoma reveals important information for the responsible physician and favours the prognostic role of minimal residual disease in neuroblastoma. Current multicenter trials in neuroblastoma trying to elucidate the relevance of residual cells at different time point during treatment are ongoing and will ultimately result in a consensus about this intruiging prognostic paramater. PMID- 15828306 TI - [The prognostic significance of P-glycoprotein in children with acute lymphoblastic leukemia and neuroblastoma]. AB - P-glycoprotein (P-gp), a pump located in the cell membrane, extrudes several clinically important drugs from the cell, and hence causes multidrug resistance (MDR). Reversing MDR is possible by using agents that inhibit the activity of P gp. In this study, we tried to elucidate the prognostic relevance of P-gp in childhood acute lymphoblastic leukemia (ALL) and neuroblastoma. In a first prospective study in childhood ALL, an immunocytochemical APAAP assay was applied. Children scoring positive had a significantly worse outcome than those with a negative test result. In a second prospective study, the prognostic relevance of P-gp was confirmed, using the combination of immunocytochemistry and a functional flow cytometric assay. The combination of immunocytochemistry with the flow cytometric functional assay is being promoted as the most sensitive and clinically relevant amongst the different techniques. In neuroblastoma, ganglioneuroblastoma and ganglioneuroma, P-gp is detected by different assays: immunocytochemistry, flow cytometric immunological and functional tests and an in vivo imaging technique using 99mTc sestamibi. Immunocytochemistry alone did not provide a prognostic role for P-gp in neuroblastoma. On the contrary, using flow cytometric tests, many neuroblastomas scored positive and a discordance was found between the expression and activity of P-gp. P-gp was found more frequently in low-stage neuroblastoma, differentiated tumours and tumours after chemotherapy. A good correlation between flow cytometric results and imaging results was seen. Consequently, 99mTc sestamibi scintigraphy is not useful as an in vivo predictor of MDR in neuroblastoma. Unlike the findings in childhood ALL, P-gp does not contribute to MDR in neuroblastoma but seems to be a marker of differentiation. PMID- 15828307 TI - Development of antibodies that interfere with the collagen-VWF-GPIb axis as new antithrombotics. AB - The epitope and the antithrombotic effect of 6B4, an antibody that inhibits GPIb, the receptor for von Willebrand Factor (VWF) on blood platelets, and of 82D6A3, an antibody against VWF that prevents the binding of VWF to collagen, were characterised. By using canine-human chimeras, alanine-scans, phage display, mutant analysis and modeling both the epitope of 6B4 in the N-terminal domain of GPIb, and of 82D6A3 in the VWF-A3 domain, could be mapped. As both epitopes furthermore are part of the ligand binding sites, this at once also explained the mechanism of the inhibition by the antibodies. Next both antibodies were tested in a thrombosis model in a stenosed artery in baboons, where they showed potent antithrombotic activities, without a noteworthy prolongation of the bleeding time. With this we thus could reveal two new strategies to prevent arterial thrombosis, which presumably may be safer than the currently available antiplatelet agents. PMID- 15828308 TI - [The key technology of picosecond time-correlated single photon counting spectrometer]. AB - The picosecond time-correlated single photon counting spectrometer developed uses the TCSPC technique and grating splitter technique that can compensate the time spread dynamically and statically. The time spread caused by the conventional optical system can be eliminated completely. The multichannel analyzer is used to measure the fluorescence decay curves and the time-resolved spectrum by opening the time windows. The method for multi-exponential fitting to the fluorescence decay is also used to deal with the data. The system theory and design of the spectrometer are presented in detail, and the process of operation is introduced with the integration of the system. Many standard samples have been measured, and the data have been analyzed and contrasted. Experimental results show that the fluorescence life-span scaled by the spectrometer reaches picosecond distinction, and the spectrometer's sensitivity is single photon counting. Time resolution of the spectrometer is 8.8 ps. PMID- 15828309 TI - [Laser resonance ionization spectroscopy of even-parity autoionization states of cerium atom]. AB - This paper describes the investigation of even-parity autoionization states of cerium atoms by three-step three-color resonance ionization spectroscopy (RIS). Twenty-seven odd-parity highly excited levels, whose transition probability is high, were used in this research. One hundred and forty-one autoionization states were found by these channels with the third-step laser scanning in the wavelength range of 634-670 nm. The ionization probabilities of different channels, which had higher cross sections, were compared. On the basis of this, eight optimal photoionization schemes of cerium atom have been given. PMID- 15828310 TI - [Investigations of transient time-dependent spectral multi-channel measurement for shocked KCl crystal]. AB - In this paper, the authors reported a new method for measuring transient time dependent spectra using multi-fiber delay coupler and optical multi-channel analysis technique. The fiber delay coupler consists of five fibers with different lengths and its time-delay between two fibers is determined by the different fiber lengths. The light beams that came from the fiber coupler were coupled by a lens system into the slit of a spectrograph and formed a spectral pattern on the focus plane of the spectrometer. A two-dimensional CCD detector converted the optical pattern into an electrical pattern, and then the transient time-dependent spectra were obtained by a personal computer. The experimental setup was constructed, and the transient time-dependent spectra were measured for KCl crystal shocked by a high speed pill. The time resolution was 20 ns, and the spectral intensity resolution reached 1/18 bit. PMID- 15828311 TI - [Study on visible chemiluminescence from reaction of microwave discharge of CF4/Ar with carbon disulfide]. AB - Using crossed molecular beam-chemiluminescence apparatus, visible chemiluminescence in the region of 300-900 nm was measured from the gas-phase reaction of microwave discharge of CF4 diluted in Ar with carbon disulfide at various pressures. The geometries and frequencies of possible reaction products were respectively investigated at the B3LYP/6-311 + + G (3df, 2pd) and MP2/6-311 + G(d, p) levels. Chemiluminescence spectrum was assigned to emission spectrum of the electronically excited states of F2S2. From the analysis of spectrum and reaction mechanism, it is proposed that carbon disulfur was dissociated by metastable Ar generated from microwave discharge. The electronically excited state of F2S2 was obtained from multistage reactions of the dissociative products with atoms F generated from microwave discharge. PMID- 15828312 TI - [Effect of Eu3+ molar ratio on the spectra of Y2O2:Eu3+, Mg2+ and Ti4+ red phosphor]. AB - Synthesis of Y2O2S:Eu3+, Mg2+ and Ti4+ red phosphor by flux fusion method was presented. The decay curve of Y2O2S:Eu3+, Mg2+ and Ti4+ red phosphor was measured and the afterglow time was over one hour. The emission spectra and excitation spectra were measured, and the effect of Eu3+ molar ratio on the emission spectra and excitation spectra were also discussed. The emission spectra showed that Y2O2S:Eu3+, Mg2+ and Ti4+ had narrow emission peaks. The emission peaks ascribed to Eu3+ ions transition from 5D(J) (J = 0, 1, 2, 3,) to 7F(J) (J = 0, 1, 2, 3, 4, 5) were found. With the increase of Eu3+ molar ratio, the emission peaks 513.6, 540.1, 556.4, 587.3 and 589.3 nm ascribed to the energy transition 5D2, 5D1 to 7F(J) (J = 0, 1, 2, 3, 4) deteriorated gradually relative to the main emission peak at 627.0 nm. The emission peaks ascribed to energy transition 5D0 to 7F(J) (J = 0, 1, 2, 3, 4) didn't weaken relative to the main emission peak. It was probably due to the so-called true activator saturation effect. This function on the higher activated states 5D2 and 5D1 was more distinct. The excitation spectra of Y2O2S:Eu3+, Mg2+ and Ti4+ showed that it had excitation peaks at 350 nm nearby, which was ascribed to the absorption of charge transfer (Eu3+-O2-, Eu3+ S2-). The excitation peaks at 468, 520 and 540 nm were ascribed to the representative energy transition 4f-4f of Eu3+ ions. With the increase of Eu3+ molar ratio, the excitation peaks 468, 520 and 540 nm strengthened relative to the main absorption peak at 350 nm nearby. PMID- 15828313 TI - [Emission mechanism in the Tb complex doped PVK system]. AB - Excitation and photoluminescence (PL) spectra of the mixture terbium complex and PVK were investigated. The excitation spectrum of Tb complex dispersed PVK film is very similar to that of the pure PVK film, indicting that energy transfer occurs from PVK to Tb complex. For the overlap between the excitation spectrum of Tb complex dispersed PVK film and the PL spectrum of PVK film is very little, the ratio of occurrence for Forester energy transfer is very little. The emission of Tb3+ mainly comes from the excited ligand, which comes from the ligand capture of electron-hole pairs. In the electroluminescence (EL) spectra, the emission of PVK is completely restrained and only emission of Tb3+ is occured, which origins from the different mechanism in comparison with photoluminescence. PMID- 15828314 TI - [Study on the relationships between chemiluminescent properties and molecular structures of some biacridine probes]. AB - The chemiluminescent (CL) properties of biacridine probes, such as 10,10' dimethyl-3,3'-disulfo-9,9'-biacridine (DMDSBA), 3,3'-disulfo-9,9'-biacridine dinitrate (DSBADN), 10,10'-diethcarboxyl-9,9'-biacridine dinitrate (DEBADN), 10,10'-dimethyl-3,3'-diamino-9,9'-biacridine (DMDABA), and 10,10'-di(4 aminobutyl)-9,9-biacridine dinitrate (DABADN), were studied in this paper, and the relationships between properties and molecular structures were obtained. A new theory and some valuable experimental data have been provided for the investigation and exploitation of the new CL probes for biomacromolecules. PMID- 15828315 TI - [A study on europium sensitized chemiluminescence of pipemidic acid]. AB - A flow injection chemiluminescence method for the determination of Pipemidic acid (PPA) was developed. It is based on luminescence produced with KMnO4-Na2S2O4 CL system sensitized by Eu3+-PPA. The familar excipients such as starch and dextrine do not cause interference. The several kinds of affect on chemiluminescence were studied. The linear range and detection limit for PPA are 7.00 x 10(-9)-9.00 x 10(-7) mol x L(-1) and 4.09 x 10(-9) mol x L(-1), respectively. This method have successfully been applied to the determination of PPA in tablet and urine. And the mechanism of chemiluminescence is proposed. PMID- 15828316 TI - [Determination of phloroglucinol by a new flow injection chemiluminescence method]. AB - Chemiluminescence reaction of phloroglucinol with potassium permanganate in the presence of formaldehyde as an enhancer was investigated. A new chemiluminescence method for the determination of phloroglucinol with flow injection technique was developed. The linear range for phloroglucinol is 5.0 x 10(-9)-5.0 x 10(-5) mol x L(-1) with a detection limit of 3.0 x 10(-9) mol x L(-1). The relative standard deviation is 2.5% for 1.0 x 10(-6) mol x L(-1) phloroglucinol in 11 repeated measurements. The method has been used to determine the content of phloroglucinol in simulated water with satisfactory results. PMID- 15828317 TI - [Preparation and characterization of 2(8-hydroxyquinoline)-2( phenol) zirconium thin film]. AB - In this paper, the new emitting material zirconium complex with 8 hydroxyquinoline and phenol (Zr(OPh)2 q2), which can form high quality nano-scale amorphous thin films at glass substrates cleaned by vacuum evaporation, was synthesized. Its crystallization, thermal stability, the structure of energy band and PL mechanism were investigated respectively by FTIR spectra, DTA-TG analysis, SEM, XRD spectra, UV-Vis absorption spectra and fluorescence spectra. Results show that the Zr(OPh)2 q2 is a thermally stable material, whose melting point is 381.2 degrees C, decomposition temperature is 467.1 degrees C, whose film can emits intense yellow-green fluorescence with peak wavelength at 525 nm and a full width at half-maximum of 107.6 nm under UV excitation (390 nm), powder has intense yellow fluorescence, therefore it may be a excellent electroluminescent material. PMID- 15828318 TI - [Research on the infrared spectrometry of aging silk fabrics]. AB - The detection of deterioration degree of ancient silk fabrics will be helpful to the selection and developing of conservation methods. This paper carried out some research on the deterioration extent and mechanism of silk fabrics by means of infrared spectrometry. The samples artificially aged and excavated from Hubei, Innermongolia and Qinghai province, were analyzed. The artificially aging was done by simulating three main natural aging factors: light, heat and hydrolysis. The infrared spectrometric analysis results show that although the infrared spectrometry is a half-quantitative analysis method, for the hydrolysis-aged silk fabrics, it can give good qualitative and better half-quantitative analysis results because of the increase of carboxyl. So the infrared spectrometric analysis is of practical value for the conservation state and aging mechanism studies of ancient silk. PMID- 15828319 TI - [Structural characterization of rare-earth stearate and its thermal stability mechanism for PVC]. AB - Structural characterization of rare-earth stearate was conducted by FTIR and XRD. The results show that the bonds between the stearate and rare-earth metal ions in rare-earth stearate are main ionic character and have the stratified crystalline structure with the crystal layer formed from a plane layer of rare-earth ions combined with two layers of fully extended zigzag chains of stearic acid radicals arranged parallell to each other on its both sides, and the rare-earth ions axes are inclined to the crystal layer planes in it. Congo red test showed that the stabilizing time increased when the stabilizers' concentration increased. The order of stability of this four rare-earth stearates is Last>Ndst>Yst>Dyst. Furthermore, the thermal stability mechanism of rare earth stearate for PVC has been presumed by FTIR. The results indicate that Last and Ndst can bate the chain reaction and exchange the labile functional groups in the backbone chains for other more stable substituents derived from the stabilizer and change the framework, while Yst and Dyst's effect is not clear. PMID- 15828320 TI - [Oil spill identification by near-infrared spectroscopy]. AB - Petroleum oil spill happens occasionally at sea. It's important to differentiate the exact products in order to carry out following actions to decrease harmfulness. In the present study, a rapid oil spill identification method by near infrared spectroscopy coupled with pattern recognition techniques is proposed. 56 simulated spilled oils of gasoline, diesel fuel and lubricating oil in marine were chosen to develop the method. Organic reagent of CCl4 was used to extract the oil. Pattern recognition techniques were established by principal component analysis (PCA) coupled with Mahalanobis' distance with the multiplicative signal correction (MSC) and Norris first derivative pretreatment. The study shows that PCA technique is a useful method to extract the main characteristics, and Mahalanobis' distance is an ellipsoidal boundary that circumscribes a data cluster. And oil spill samples with concentration above 0.4 microL x mL(-1) can be successfully identified by the method. The developed technique could be further applied to the identification of spilled oil in marine. PMID- 15828321 TI - [A novel method for the determination of inorganic ions in complex plant samples by near infrared spectroscopy]. AB - Inorganic ions in plant samples can be determined by near-infrared (NIR) spectral technique, because they are combined with organic groups with NIR absorption. A method for the determination of inorganic ions in complex plant samples was established based on the combination of discrete wavelet transform (DWT) and NIR technique. In the proposed method, the raw NIR data and their wavelet coefficients are used for modeling and prediction of the contents of potassium in tobaccos by partial least square method (PLS). It is shown that there is almost no loss of spectral information with the NIR data compressed to 3.3% of its original size. The model based on wavelet coefficients is better than that based on the full NIR spectral range. With the improved method, accurate prediction can be achieved. PMID- 15828322 TI - [FTIR study on the finger nail with carcinoma of nasopharynx]. AB - Finger nails from twenty normal people and three with carcinoma of nasopharynx were studied by Fourier Transform Infrared (FTIR) technique. The results showed that there are obvious differences between FTIR spectra of them in spectral parameters such as frequency, intensity and band shape etc. The most striking differences in the spectra were observed in the change of amide II, the disappearance of delta(s) (CH3) peak, and at 874.0 cm(-1) whether appeared an absorption peak. The changes involving the phosphate symmetric stretching nu(s, PO2-) and asynmmetric stretching nu(as, PO2-) modes, the CH3 and CH2 groups stretching (v(s, CH2), nu(as, CH3)) bending (delta (CH3)) modes and the C-O stretching nu (C-O) mode were discussed. In addition, the changes of structure of hydrogen-bonding of nucleic acid and cell proteins and the packing and the conformational structure of the membrane lipids were analysed. The average wave number of band of nu(s) (PO2-) shifted from 1080.0 to 1077.6 cm(-1) and that of nu(as) (PO2-) shifted from 1239.4 to 1238.4 cm(-1), which indicated that the degree of hydrogen-bonding formed by oxygen atom of the phosphodiester groups of nucleic acids was weakened. The average wave number of band of delta (CH2) of membrane lipids shifted from 1453.1 to 1453.7 cm(-1), and its peak intensity was slightly enhanced, which suggested that the conformational structure of the methylene chains of membrane lipids is more disordered than in normal nail. PMID- 15828323 TI - [Influence of smooth, 1st derivative and baseline correction on the near-infrared spectrum analysis with PLS]. AB - This paper studied the influence of using pre-procession such as smooth, 1st derivative and baseline correction on the analysis of near-infrared spectrum. Comparing the analysis results by the pre-procession methods, and using PLS arithmetic, the best pre-procession was determined. In smooth pre-procession method, the best smooth points were proposed for regression using PLS. The analysis result is satisfactory. PMID- 15828324 TI - [Wavelet denoising and optimization of two-dimensional correlation IR spectroscopy]. AB - Wavelet analysis and wavelet denoising was applied to analyse the data of two dimensional correlation IR spectroscopy of traditional Chinese medicine for the first time in this article. The results showed that, after denoising, the authors can decrease the interference to the synchronous spectra of two dimensional correlation IR spectroscopy effectively, and tone up the signal peaks and separate the peaks which overlap together before denoising. In this way, the authors can not only improve the spectral revolution, but also get more spectral informations. PMID- 15828325 TI - [Identification of an unknown assistant blowing agent in AC blowing system]. AB - Recently it has become a trend to modify AC by adding different additives in AC system. In the modification, it is a key question to know what kind of additives should be added. So, qualitatively deformulating the additives will certainly help the research on and development of modifying AC a lot. In this paper, TLC (Thin Layer Chromatograph) was used to separate and purify the unknown assistant blowing agent in AC blowing system. After that, FTIR-Microscope was used to indicate the functional group in the uncommon unknown compound. Through combination of FTIR analysis, elemental analysis and other physical and chemical analysis, the molecular structure of the unknown was deduced. Finally, by explaining the unknown compound's MS spectra, the molecular structure of the unknown was confirmed further. The method can be extended to analyze other additives in AC blowing agent system whose various standard spectra are absent. PMID- 15828326 TI - [A DRIFTS study of SO2 oxidation on the surface of CaCO3 particles]. AB - CaCO3 is an important component of mineral dust aerosols in the atmosphere, but its role in atmospheric heterogeneous chemistry is still unclear. In this paper, the oxidation of SO2 on CaCO3 particles was studied using diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). Formation of sulfite and sulfate was identified on the surface, and the role of O3 on the oxidation process is determined by comparing the experiments in different conditions. DRIFTS is proved as a useful tool for studying heterogeneous reactions on CaCO3 particles. The results show that in the presence of O3, SO2 can be oxidized into sulfate on the surface of CaCO3 particles. A two-step mechanism that involves adsorption of SO2 followed by oxidation is presented. PMID- 15828327 TI - [Extraction and analysis of sargassum hemiphyllum polysaccharides]. AB - Sargassum hemiphyllum polysaccharides (SHP) was extracted by thermal water method and the physical and chemical characters, the extraction rate, contents, compositions of SHP was studied. The result showed that SHP was ashen powder, water-soluble, insoluble in organic solvents. The reaction of iodide-potassium iodide was negative to evaluate that SHP was nonstarch polysaccharides. Extraction rate was 7.04% and the content of polysaccharides was 82.9%. Ultraviolet spectrum showed that there were little DNA and protein. Infrared spectroscopy showed that SHP was main pyrano polysaccharides and had beta-linked glycopolysaccharides in molecule structure of SHP. Thin layer chromatography traced that SHP could be xylan. The results indicated that the extract was not only polysaccharides but also higher purity and the method was high efficiency. PMID- 15828328 TI - [Preprocessing of near infrared spectroscopic data]. AB - Spectroscopic data of samples recorded by spectroscopic instruments are confused by a series of noises, and interferences, therefore the proper data preprocessing is the basis of the following spectroscopic calibration, model establishment and transference, which is very important for the achievement of accurate analytical results. This paper reports our research work, combined with NIR spectroscopic analyses of the protein contents of wheat, that is the preprocessing of NIR spectral data recorded for 66 different wheat samples by a NIR grating spectrophotometer and a NIR Fourier transform spectrometer, respectively. The preprocessing algorithm is wavelet transform with the Gaussian first and second order derivatives. Compared with the result of preprocessing by normal first order difference algorithm, the wavelet transform algorithm by Gaussian derivatives was proved to be very effective and applicable, the spectra were smoothed perfectly, noises were eliminated obviously, and the spectral sections, which include all useful information for spectral analyses, were displayed clearly. So, it is very beneficial to the following spectral calibration, model establishment and transference. PMID- 15828329 TI - [Adopting the method of principal components analysis combined with correlation coefficient to increase the predicted concentration's accuracy of benzene and its homology mixture]. AB - The concentrations of benzene and its homology mixture were measured by near infrared spectra, and the emphasis was put on the character of the principal component and its physical significance. It is pointed out that the anterior principal components are very similar to the correlation coefficient of the multi component solution and the theoretical proof for the right condition is given. The high frequency noise of the system can be checked out by principal component combined with the correlation coefficient. Removing the noise can greatly increase the accuracy of the prediction model. PMID- 15828330 TI - [Study on the synthesis, characterization of structure and fluorescence properties of the solid complex of europium ion with 1,10-phenanthroline and 2 thiopheneglyoxylic acid]. AB - The complex of europium with 1,10-phenanthroline and 2-thiopheneglyoxylic acid (HL) was synthesized. Its chemical formula was determined to be [EuL2 x phen x (H2O)3]NO3 by elemental analysis, electrical conductivity, IR and 1H NMR. It was found that the europium ion coordinated to the O atoms of 2-thiopheneglyoxylic acid and H2O, and to the N atoms of 1,10-phenanthroline. Its fluorescence property was determined by the excitation and emission spectra at room temperature, the emission peaks at 592 and 618 nm were assigned to the 5D0-7F1 and 5D0-7F2, respectively. It was also observed that the title complex emits strong red fluorescence under ultraviolet light at room temperature. In the IR spectrum, the characteristic absorption nu(C=O) (1719 cm(-1)), nu(C-O) (1232 cm( 1)), delta(O-H) (909 cm(-1)) of 2-thiopheneglyoxylic acid disappeared in the complex, the nu(as) and nu(s) of -COO- appeared at 1642 and 1408 cm(-1) in the complex respectively. In 1H NMR, the signals of 2-thiopheneglyoxylic acid were shifted upfield, and those of 1,10-phenanthroline downfield. It is proved by the TG curve that the complex is stable from ambient temperature to 250 degrees C. PMID- 15828331 TI - [Study of the factors effecting surface-enhanced Raman scattering reporter labeled immunogold colloids]. AB - The applications of the high sensitivity of Surface-Enhanced Raman Spectroscopy (SERS) in labeled immunoassay have proved to be very useful. In the sandwich structure of "immobile antibody-antigen-labeled antibody", the authors prepared the labeled antibody by attaching gold nanoparticles, which was first labeled with molecule that had SERS signal, to the antibody. In this article, colloids were labeled with Raman reporter thiophenol through S-Au bond on gold and formed surface-enhanced Raman scattering (SERS) reporter-labeled gold colloids. Then, goat anti-rat antibody was adsorbed onto the reporter-labeled gold colloids and formed reporter-labeled immunogold colloids. This paper studied factors affecting FT-SERS reporter-labeled immunogold colloids such as the sizes of gold particles, the amount of thiophenol in gold colloids and processing time, and whether goat anti-rat antibody would influence the SERS of reporter-labeled gold colloids. PMID- 15828332 TI - [Raman spectroscopy study of 1,3-dimethyluracil and its ab initio (DFT) calculation]. AB - The Raman spectrum of polycrystal 1,3-dimethyluracil (DMU) was obtained and investigated. Ab initio (DFT) calculation (B3PW91/6-31G method) of DMU was performed. And the vibration modes of DMU molecular were analyzed. The results suggest that DMU molecule, as a model compound of bio-base, has some advantages in Raman analysis, such as stronger intensity of marker bands and sharper, clearer bands of double band stretching than uracil's. The ultimate goal of this research is to get large quantity of spectral information of DMU and its cyclobutane dimers, use them as model compounds to research on the Raman characteristics of DNA's or RNA's photodamage. And this work is also an essential supplement to the 1,3-dimethyluracil's Raman study. PMID- 15828333 TI - [Raman spectroscopic studies on methanol under high pressure]. AB - This paper studies Raman spectra of methanol atpressures from 169.2 MPa up to 713.8 MPa and at ambient temperature. The result shows that the spectral profile of stretching vibrational modes of CH and OH gradually changes and Raman shift moves to higher and lower frequency respectively with increasing pressure, which indicates that the energy of C-H bonding increases with pressure, and the influence of hydrogen bonding on O-H function group is greater than that of pressure. In addition, the relative peak area of O-H stretch band decreases with pressure, showing that the total intensity of O-H bonding is weaker compared with C-H bonding. PMID- 15828334 TI - [Theoretical study on the structure and spectra of C72O3]. AB - The optimized geometries and stability of eight different isomers of C72O3, the five-membered ring derivatives of C70 fullerenes, were investigated by using AM1 method. According to the optimized geometries of eight different isomers, AM1 and ZINDO/Cl method were employed to study the vibrational spectra and electronic spectra, respectively. The calculated results indicate that the -C2O3 groups are added preferably to the seats of CI-CII and CIII-CIII bond to form the isomers of A and B. The additive C-C bond is elongated to become primarily a single bond. The stability of isomer B is very similar to that of isomer A that has been prepared experimentally. The calculated vibrational spectra of isomer A, similar to those of isomer B, are consistent well with the results from the experiment. The electronic transitions of isomers of C72O3 were assigned and the red shift of electronic spectra was discussed. The spectra of other isomers were predicted theoretically. PMID- 15828335 TI - [Investigation of electron paramagnetic resonance spectrum in Ni2+:RbMgF3]. AB - The optical spectra, the zero-field splitting parameter D and the g-factor of transition-metal ions in crystal are related to the crystal structure. A method, which can be used to calculate accurately the optical spectra, the zero-field splitting parameter D and the g-factor, was founded by using the parametrization d orbital for Ni2+, the energy matrix elements of strong-field coupling scheme for d2 (d8) electron configuration in trigonal crystal field and by adopting the method of complete diagonalization. In the calculation, all spin singlet states and spin triplet states have been considered, and then the result is exact. The present theory shows that g(parallel) > g(perpendicular), D < 0 or g(parallel) < g(perpendicular), and D > 0 are due to the different distortion of ligands along C3 axis. The electron paramagnetic resonance spectrum in Ni2+:RbMgF3 with C3v and D3d symmetry was calculated by using the theory. The relationship between the EPR spectrum in Ni2+:RbMgF3 and the crystal structure were investigated. The agreement of the theory with the experiment shows that the present theory is reasonable. PMID- 15828336 TI - [The medium effects of fluorescence of eosin-5-isothiocyanate as a luminescent marker]. AB - In this paper, the medium (pH, organic solvent and surfactant) effects on fluorescence of eosin-5-isothiocyanate (eosin-ITC) have been studied in detail. The results show that the pH and polarity of system studied are two great important influence factors on fluorescence of eosin-ITC. The fluorescence intensity of eosin-ITC in weak acidic medium is higher obviously than that of in weak basic medium, but it is sensitive to change of pH, be careful to use. The reaction between surfactant and eosin-ITC shows a good electric selectivity, and the effect of CTMAB on fluorescence of eosin-ITC can be classified as two stages: monomer quenching (below cmc) and micelle sensibilization (above cmc). In the presence of micelle of CTMAB, the fluorescence intensity of eosin-ITC in weak basic medium is even higher than that of in weak acidic medium. In the presence of high organic solvent content, the fluorescence intensity of eosin-ITC is also increased obviously. These properties are available for use. PMID- 15828337 TI - [Study of solid-phase time-resolved fluorescence label immunoassay]. AB - This paper describes optimal conditions for HBsAbIgG labeling with a new fluorescence probe, 4,7-bis-chorosulfophenyl-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) for the solid phase time-resolved fluorimmunoassay (TRFIA). The result of experiment under states clearly that BCPDA may react with protein under relative mild condition. The relative bioactivity of reacted protein was more than 80%. The labeling molar ratio of BCPDA for HBsAbIgG was 45-70. The recovery was higher than 80%. Protein-BCPDA-Eu3+ complex is stable. It can emit very high fluorescence intensity with very long fluorescence life times. The fluorescence of Protein-BCPDA-Eu3+ complex has a very large stokes shift (270 nm). The emission band at 611.2 nm is very narrow. The research provides the base for developing non-isotopic immunoassay technique and clinical medical diagnosis. PMID- 15828338 TI - [Ion fluorescence probe detecting the formation of triplex DNA]. AB - The fluorescence of Tb3+ is not quenched after the rare earth ion is combined with DNA. The fluorescence intensity is related not only to the kinds of bases of DNA but also to the kinds of DNA. The rare earth ion Tb3+ was used as the fluorescence probe to detect the formation of triplex DNA. The results show that the fluorescence intensity of Tb3+ combined with polydA is much stronger than that of Tb3+ combined with polydT, which testifies that the fluorescence intensity is related to the kinds of bases combined with Tb3+. The results also demonstrate that the rare earth ion can be used to detect those three forms of DNA though their fluorescence peak positions are similar when Tb3+ as a fluorescence probe is combined with single strand DNA (ssDNA), double helix DNA (dsDNA), and triple helix DNA (tsDNA), respectively. However, their intensities are quite different. The fluorescence intensity of Tb3+ combined with ssDNA-Tb3+ is the largest. And the fluorescence intensity of Tb3+ combined with tsDNA-Tb3+ takes the second place. The minimum peak intensity belongs to the contribution of Tb3+ coupled with dsDNA-Tb3+. The reason is that the capability of energy transference of ssDNA, dsDNA and tsDNA is different. The different degree of energy transference influences the intensities of Tb3+. The influences of pH and the metal ion on the formation of triplex DNA were also studied. The authors found that the neutral pH and high valence metal ion are beneficial to the formation of ts-DNA. PMID- 15828339 TI - [Fluorescence spectra characters of nanostructured gold thin-film]. AB - Gold colloid was prepared via electrochemical method. Nanostructured gold thin films were deposited on glass by using self-assembled technique. UV-visible spectra and fluorescence emission spectra were measured at room temperature. The absorption spectrum features show two peaks and the longer wavelength one at 610 nm corresponds to linear aggregated gold nanochains. The fluorescence emission peaks at about 375 nm and 700 nm are due to the local field enhancement via coupling to the transverse and longitudinal surface plasmon resonance from gold nanochains. Both the intensity and number of the fluorescence emission peaks increase with increasing excited light and the layers number. It is indicated that the nanostructured gold thin-films formed a disordered particles system. Further more, multiple scattering takes place in these self-organized random micro-cavities. PMID- 15828340 TI - [Fluorescence reaction of terbium(III) ion and norepinephrine and its analytical application]. AB - Experiments indicated that terbium(III) ion could form complex with norepinephrine, then emitted its characteristic fluorescence. In the present paper, the fluorescence reaction of terbium(III) ion with norepinephrine was studied in detail. Thus a new method for the determination of norepinephrine was propsed. The fluorescence intensity was measured in a 1 cm quartz cell with the excitation and emission wavelengths of 300 and 545 nm, respectively. The result showed that the fluorescence intensity of the system presented a linear relationship with the concentration of norepinephrine in the range of 0.01-50 microg x mL(-1), and the detection limit (S/N=3) was 1.0 ng x mL(-1). The method was used to determine pharmaceutical sample which contained norepinephrine, and the result was satisfactory. PMID- 15828341 TI - [The fluorescence characteristics of norfloxacin and its application]. AB - The fluorescence characteristics of norfloxacin in different pH media have been studied by spectrofluorimetry. It was found that in the neutral and alkalescent media the fluorescence signal of norfloxacin was weak, while in the acidity media the signal was very strong. The maximum fluorescence signal of norfloxacin could be obtained at pH 3.04, and the norfloxacin fluorescence emission wavelength has red-shifted 30 nm compared with the other condition. Based on this, a spectrofluorimetric method of the determination of norfloxacin in pharmaceuticals is proposed. The linear range was 0.016-0.96 microg x mL(-1), the detection limit was 0.016 microg x mL(-1), the RSD was 1.65%-2.86%, and the recovery was within the range of 99.7%-100.94%. At pH 3.04, norfloxacin in human urine can be well resolved by Synchronization-1st derivative fluorescence spectroscopic techniques with delta lambda = 90 nm, and the interference from urine blank can also be eliminated. The linear range was 0.096-3.19 microg x mL(-1) for determining norfloxacin in human urine sample. The detection limit was 0.096 microg x mL(-1), the recovery was within the range of 96.24%-104.4% and RSD was 0.71%-2.60%. Pharmacokinetic study of norfloxacin showed that norfloxacin reached its peak concentration in urine samples two hours after a healthy man took capsules. The method is simple, rapid and accurate with high sensitivity and good selectivity, and suitable for the determination of norfloxacin. The method was applied to analysis of norfloxacin in drugs and biological samples. PMID- 15828342 TI - [Study on the interaction of cystine and polyphenol oxidase from nicotian tobaccum]. AB - The interaction of cystine and polyphenol oxidase (PPO) from nicotian tobaccum has been studied. The results show that cystine has an inhibitory effect on the enzymatic activity, the mechanism of which might be that the thiolether in cystine coordinates with Cu2+ in the active site of PPO, and the inactivation constant value of PPO by cystine is 0.633 min(-1), while the substrates or the products inhibit their combination. Cystine can also be combined with the products of enzymatic reaction to form a colourless compound. Cystine can inhibit the enzymatic activity completely when the molar ratio of cystine to PPO approaches 16,000:1. The effect of cystine on the fluorescence changes with the molar ratio of cystine to the PPO. However, when the molar ratio gets to 75:1, cystine has no longer the effect on either the fluorescence spectra or the synchronous spectra. Microenvironment of trp residue in PPO is more hydrophobic than that of free trp in water. PMID- 15828343 TI - [Fluorescence spectroscopy determination of lomefloxacin by charge transfer complex formation with chloranilic acid]. AB - Charge-transfer complex was formed between LMX as the donor and Chloranilic Acid (CL) as the acceptor has been studied by fluorimetry. It was shown that the n-pi complex can be formed at ambient temperature, which can emit, which strong fluorescence. Different variables and parameters affecting the reactions were studied and optimized. Based on this, a simple and reliable fluorescence spectroscopy method for the determination of Lomefloxacin (LMX) has been developed. Interference from some co-formulated drugs was also studied. No interference was observed due to additives commonly present in the pharmaceutical preparations. The proposed methods could be applied successfully to the investigated pure compounds and pharmaceutical dosage forms with good accuracy and precision. The linear range is 0.04-0.8 mg x L(-1), the detection limit is 0.04 mg x L(-1), the recoveries of LMX are 97.4%-99.3%, and RSD is 1.3%-2.6%. Finally, the charge-transfer reaction mechanism was discussed. The composition of the change-transfer complex was found to be 1:1 by Bent-French and curved intersection methods. This ratio may be due to the presence of the fluorine atom acting as an electron drawing group in the molecule of lomefloxacin. The benzene ring has lower electron density, but nitrogen atom in 4' of piperazingl has higher electron density and is less sterically hindered. So n-pi charge transfer complexes were formed. PMID- 15828344 TI - [Photobleaching properties of GFP expressed in human lung cancer cells]. AB - The characteristic properties of GFP make this protein a good candidate for use as a molecular reporter to monitor patterns of protein localization, gene expression, and intracellular protein trafficking in living cells. In this study, the dicistronic expression vector (pEGFP-C1) was used to be transfected into human lung cancer cell line (ASTC-a-1) and a positive clone which stably expressed GFP in high level was obtained. The results showed that the green fluorescent protein expressed in tumor cells was also photobleached under intense irradiation (-488 nm) and the degree of photobleaching varied with the intensity of the excitation. In order to analyze the effect of low temperature on the fluorescent sample, the photobleaching of tumor cell slice stored at -20 degrees C was observed additionally. PMID- 15828345 TI - [Absorption spectra study of cation surfactant-[AuI4]- association particle system and its analytical application]. AB - In 0.01 mol x L(-1) HCl medium, there is an absorption peak at 350 nm for [AuI4] . The association particle system of CTMAB-[AuI4]- exhibits violet-red color. It has a new absorption peak at 520 nm. The CTMAB concentration cCTMAB in the range of 0-7.0 x 10(-5) mol x L(-1) is linear to the A(520 nm) value. Its regression equation is A(520 nm) = 0.989 x 10(4) cCTMAB + 0.0138, regression coefficient is 0.9995, and the mole absorption coefficient is 1.059 x 10(4) L x mol(-1) x cm( 1). A new spectrophotometry has been proposed for the determination of cation surfactant. The resonance scattering spectra show that the CTMAB+ and [AuI4]- combine to form (AuI4-CTMAB) association molecule with hydrophobicity by means of attraction forces. The association molecules automatically aggregate to form (AuI4-CTMAB)n association particles. So, the particles exhibit two resonance scattering peaks at 580 and 470 nm, and the system exhibits violet-red. PMID- 15828346 TI - [Spectral properties of the association particle system of quinine-[PtI6]2- and its analytical application]. AB - In 0.01 mol x L(-1) HCl medium, red color [PtI6]2- and quinine combine to form association molecule. The molecules aggregate automatically owing to strong hydrophobic and molecular forces. And PtI6-quinine association particles with violet-red color formed. It exhibits three resonance scattering peaks at 310 nm, 400 nm and 610 nm, and a Rayleigh scattering peak at 470 nm. The absorption values all increase in the wavelength range of 350-740 nm, and the fluorescence peak at 450 nm was quenched. Under the conditions chosen, the quinine concentration in the range of 0-40 x 10(-6) mol x L(-1) is linear to the A(620 nm) value. The mole absorption coefficient epsilon(620 nm) is 1.31 x 10(4) L x mol(-1) x cm(-1). The results show that the association particles produce the resonance scattering effect, fluorescence quenching and the violet-red color. PMID- 15828347 TI - [The spectroscopic behaviour and extraction separation of Cu(II), La(III), U(VI) and Ce(IV) in two-phase aqueous systems]. AB - In polyethylene glycol 2000 (PEG)-(NH4)2 SO4-extractant (sodium diethyldithiocarbamate, DDC) two-phase aqueous systems, spectroscopic behaviour of complexes of Cu(II), La(III), U(VI) and Ce(IV) with DDC as the extractant in PEG phase and general water phase was investigated. Comparing to DDC spectrophotometry, the new method has the advantage that the maximum wavelength of complex of Cu(II) shifts to red wavelength. Extraction mechanism and existing form of complexes of metal ions in PEG phase were studied. Effects of different acidity, quantities of DDC and salt, different surfactant on the extraction efficiency of copper(II), lanthanum(III), uranium(VI) and cerium(IV) were discussed. Experimental results show that Cu(II) was almost completely extracted in PEG phase at pH 5-8, U(VI) was partly extracted at pH 1-11, while La(III) and Ce(IV) were not extracted at pH 3-6 buffer solution. So, extraction separation of Cu(II)-La(III) and Cu(II)-Ce(IV) was carried out by controlling the solution acidity. PMID- 15828348 TI - [Absorption spectra study of the association nanoparticle system of berberine [AuI4]- and its analytical application]. AB - In 0.008 mol x L(-1) HCl medium, [AuCl4]- and I- combine to form [AuI4]-. [AuI4]- reacts with berberine (BB) to form (AuI4-BB) association molecule. The molecules aggregate automatically owing to strong hydrophobic and molecular forces. And (AuI4-BB)n association nanoparticles with orange-yellow color were observed. It exhibits a resonance scattering peak at 520 nm. The absorption peak for the nanoparticle system and the [AuI4]- is at 350 nm. However, the absorbance values for the nanoparticle system increased in the visible light region. Under the conditions chosen, the BB concentration in the range of 0.8 x 10(-6) mol x L(-1) 20 x 10(-6) mol x L(-1) is linear to the A(450 nm) value. The results show that the formation of (AuI4-BB)n nanoparticles produces the resonance scattering effect and the orange-yellow color. PMID- 15828349 TI - [Extraction catalytic spectrophotometric determination of trace chromium by synergetic catalytic effect]. AB - In this paper, a new synergetic extraction catalytic spectrophotometric method for the determination of trace chromium was studied. The method is based on the iron(III) and chromium(VI) synergetic catalyzed oxidation of o-aminophenol by hydrogen peroxide in weak acidic medium at pH 5.5. The reaction time, concentration of o-aminophenol in aqueous phase, and degree of reaction were controlled by extraction equilibrium. The absorbance of organic phase was measured at 424 nm. The linear range of the method is 0.0010-0.90 mg x L(-1). The detection limit for chromium(VI) is 5.0 x 10(-6) g x L(-1). The relative standard deviation (n = 11) for 0.20 microg x mL(-1) level of Cr(VI) is 2.0%. The method has been applied to the determination of chromium in tap water and waste water with satisfactory results. PMID- 15828350 TI - [A catalytic kinetic spectrophotometric method for determination of trace zinc(II) in Mongolian medicine and hair sample]. AB - A new catalytic kinetic spectrophotometric method for the determination of trace zinc(II) was developed, which is based on the decolour reaction of H2O2 with calcon catalyzed by zinc(II) in weak acid medium. The optimum condition and kinetic property of the reaction were also studied. The detection limit in this method is 0.68 microg x L(-1). The apparent activation energy Ea and the apparent rate constant R are 45.1 kJ x mol(-1) and 5.80 x 10(-3) s(-1) respectively. The range of determination is 0.80-40.00 microg x L(-1). The method has been applied to the determination of zinc(II) in Mongolian medicine and hair sample with satisfactory results. PMID- 15828351 TI - [Study on spectral properties of Chinese herbal medicines additives in cosmetic]. AB - Chinese herbal medicines as additives in cosmetic can both keep the properties of cosmetic and have maintaining, health protection and remedial effect. Some of the Chinese herbal medicines can absorb ultraviolet and cure sunburn. This article mainly studies the spectral properties of several kinds of Chinese herbal medicine additives in cosmetic by ultraviolet and visible spectrophotometer, and analyses the ultraviolet absorption of the Chinese herbal medicine additives. It was shown that gynostemma pentaphyllum can absorb ultraviolet very well. It can be a promising natural additive in the health protection cosmetic. Codonopsis pilosula, floss chrysanthemum indicum, radix scutellariae and radix glycyrrhizae can absorb ultraviolet. They can also be the additives, while ligusticum wallichii can't absorb ultraviolet, so it can not be used as the sunburn protection additive. PMID- 15828352 TI - [Determination of trace manganese in Mongolian medicine by methylthymol blue Mn(II)-H2O2 catalytic system]. AB - A new catalytic kinetic spectrophotometry method for the determination of trace manganese based on the oxidation reaction of H2O2 with methylthymol blue in Na2B4O7-NaOH medium has been developed. The range of the determination range was 0.20-40.00 microg x L(-1). The apparent activation energy and apparent rate constant were 73.24 kJ x mol(-1) and 5.62 x 10(-4) x s(-1), respectively, and the detection limit was 6.4 x 10(-10) g x mL(-1). The method has been used to determine trace manganese in Mongolian medicine with satisfactory result. PMID- 15828353 TI - [Study on optimizing the analysis condition for active compound in Eucommia olive]. AB - The absorption spectroscopic determination of aucubin by condensation with p dimethylaminobenzaldehyde was optimized in this paper. A high sensitive method was established and the linearity range of the method was from 4.5 to 45 microg x mL(-1). The influences of the factors, such as the dosage of coloring agent, reaction time, heating temperature and the dose of acid etc., on the system were studied. And the result indicated that all of the factors had obvious effects on the system. The method has been applied to determining the content of aucubin in the extract of Eucommia olive with satisfactory results. PMID- 15828354 TI - [Resonance light scattering of rhodamine B]. AB - The characteristic and mechanism of resonance light scattering (RLS) of Rhodamine B (RhB) were studied. In acidic solutions with pH from -0.38 to 4.10, the intensity of RLS increases with an increase in pH and reaches a maximum at nearly neutral pH. The change in RLS intensity with the wavelength was not accordant with Rayleigh scattering law. The fluorescence excitation and emission spectra of RhB overlap partly, and the RLS peak lies between the fluorescence excitation and emission peaks. In the three-dimensional fluorescence contour spectra of RhB, Rayleigh scattering line intersects fluorescence contour. In light polarization experiment, the polarization of RLS was measured to be P approximately = 0.1. All the above experimental facts reveal that the RLS of RhB is mainly resonance fluorescence. The mechanism of RLS enhancement by the increase in pH is that the formation of fluorescence species occurs in acid-base equilibrium. The RLS peak of RhB appears within the envelope of absorption, and light scattering is affected by light absorption, so, the relationship between RLS intensity and RhB concentration is not strictly linear. PMID- 15828355 TI - [Study on the determination of metal elements in wild jujube seed decoction by ICP-AES]. AB - The determination of metal elements in Chinese medicine wild jujube seed decoction was studied by ICP-AES after HNO3-H2O2 digestion. The average recoveries of the method for Ca, Cr, Al, Fe, K, Mg, Mn and Zn in wild jujube seed decoction were found to be 95.5%-106.8%, while RSD (n=5) was found to be 1.7% 3.9% for these metal elements. The method was applied to the analysis of some practical samples and the results obtained were satisfactory. PMID- 15828356 TI - [Studies on the determination of silicon and phosphorus in ferromanganese by ICP AES]. AB - An ICP-AES method for the determination of silicon and phosphorus in ferromanganese was studied. Digesting samples by microwave and all kinds of parameters of the analytical method were optimized, which included selecting the acid and controlling the temperature and pressure of dissolving the sample, confirming analytical spectrum, considering the effect of sample matrix, analytical pH and disturbing elements on the determination results. By optimizing and choosing all kinds of condition parameters, the simultaneous determination of silicon and phosphorus was realized and satisfactory results were obtained. The linear correlation coefficients of Si and P were 0.9998 and 0.9996, respectively. The detection limit for silicon was 0.0060% with recovery of 97.0%-101%. The detection limit for phosphorus was 0.030%. This method is accurate and quick with less reagent dosage and broa linear range. The way of this determination is fit for the determination of silicon and phosphorus with low, middle and high carbon contents. PMID- 15828357 TI - [Spectral determination of the contents of mineral elements in polygonum cuspidatum]. AB - The present paper reports the determination of mineral elements by FAAS, including K, Ca, Na, Mg, Fe, Zn, Mn and Cu. The best condition for the determination was investigated. The best ionization inhibitor and acidity media were confirmed. The experiment used CsCl to prevent ionization interference, and the concentration of HCl was controlled below 2%. At the same time SrCl prevented interference of P to Ca. So the value of absorbance was stable. The results were obtained by calibration curve method. The analysis method is simple, rapid, accurate and fitting to the determination of mineral elements in plants. The recoveries of the method are 90.5%-108.2%. The relative standard deviations are 0.3%-0.7%. The result show that large amounts of mineral elements are embodied in polygonum cuspidatum. The contents of K and Ca are in conformation with the medical effects of polygonum cuspidatum, and each element is related closely to its growing. PMID- 15828358 TI - [Determination of trace zinc, manganese, cadmium and lead in aloe by microwave digestion atomic absorption spectrometry]. AB - A microwave digestion procedure was developed for the determination of trace Zn, Mn, Cd and Pb in aloe-leaf cuticle and aloe-leaf gelatin, using the obturated vessel microwave digestion system with a pressure controlling part, and the amounts of these trace metallic elements were determined by atomic absorption spectrometry after microwave digestion. The effects of the composition of digestion solution, the ratio of the sample to digestion solution, and the digestion time were studied. It is satisfactory to apply the microwave digestion procedure to the determination of Zn, Mn, Cd and Pb under the optimized condition with the recovery of 95.0% to 110.0% and RSD of 0.3% to 6.2%. The results show that this method is rapid and simple with low environmental contamination and complete digestion of the sample. PMID- 15828359 TI - [Recent advance of resonance light scattering technique for the determination of nucleic acids]. AB - Nucleic acid analysis is one of the most important techniques for the study in biotic field. The spectrophotometry using inherent ultraviolet absorption spectrum of nucleic acids and the spectrofluorimetry based on the interaction of nucleic acids and fluorescence probes were usually used for nucleic acid analysis. However, the spectrophotometry was severely limited by its low sensitivity, and the spectrofluorimetry suffered from the disadvantages of high cost and toxic reagents. Recently, resonance light scattering has emerged as a new nucleic acid analysis technique. The resonance light scattering analysis is becoming increasingly important because high sensitivity can be obtained with a common spectrofluorimeter by using inexpensive and safe reagents. In this paper, the principle of resonance light scattering was briefly stated and the recent advances of the nucleic acid analysis by resonance light scattering were reviewed with 57 references. The resonance light scattering probes for nucleic acid analysis were divided into four groups: organic dyes; cation surfactants; metal ions and their complexes; and drugs. The light scattering analysis for nucleic acids based on the formation of large particles was also stated. PMID- 15828360 TI - [The application of variable theoretical alpha influence coefficent to the matrix correction in Cr-Fe-Ni stainless steel]. AB - X-ray fluorescence analysis software Spectra Plus was used to calculate theoretical alpha influence coefficents of other elements to Cr in seven stainless steel standard samples, theoretical alpha influence coefficients of elements, by which Cr signal was enhanced, varied largely with the change of elements content. Variable theoretical alpha influence coefficients, which varied with elements content, were used to correct the matrix effects in stainless steel, the secondary excitation of Cr by other elements were corrected, and Cr (0.3%-20.8%) in stainless steel and low alloy steel could be analysed in accordance with one calibration curve. The matrix effects in samples can be corrected by variable theoretical alpha influence coefficients, so the measurable content range of calibration curve was enlarged. The contents of fifteen elements Al, Si, P, S, Ti, Cr, Mn, Co, Ni, Cu, As, Mo, Sn, W and Pb in stainless steel were measured by X-ray fluorescence spectrometer, variable theoretical alpha influence coefficients were used to correct the matrix effects, and the analysis results are comparable to those obtained by wet chemical method. PMID- 15828361 TI - Readers respond. Agreeing to disagree. PMID- 15828362 TI - Understanding the obesity epidemic: what does the lab need to know? PMID- 15828363 TI - Pharmacogenomics and its applications. PMID- 15828364 TI - Survival of the fittest. PMID- 15828365 TI - "Green thumb" not welcome in the lab. PMID- 15828366 TI - Can public policy curtail obesity? PMID- 15828367 TI - [The management of osteoarticular problems by internal/general practitioners is inadequate. Is this a problem?]. PMID- 15828368 TI - [Clinical evaluation of rheumatoid arthritis in practice]. AB - Clinical evaluation is an integral part of medical practice. However, recent data have demonstrated that a systematic and standardized evaluation modifies the prognosis of our rheumatoid arthritis patients. The systematic use of activity indexes allows us to better appreciate the needs of our patients and the necessity to optimize and intensifie treatment. Likewise, auto-evaluations tools bring useful information to patient management. PMID- 15828369 TI - [Interest of magnetic resonance imaging in rheumatoid arthritis]. AB - Magnetic Resonance Imaging (MRI) is the best imaging method in early detection and management of rheumatoid arthritis (RA). There are other imaging methods available as ultrasound, scintigraphy, computed tomography and plain radiography for imaging RA but MRI provides the best sensitivity in detecting inflammatory changes in the joints. MRI shows the best intra and interobserver reliability and low variation between repetitive examinations. MRI has shown the best visualisation and the greater sensitivity to detect erosion in early RA, compared to standard radiography. The use of a contrast agent further increases the sensitivity in detecting erosions and differentiate synovial proliferation from fluid collections. Otherwise, intraossous cyst, tenosynovitis, bone marrow edema, that are concomitant manifestations of the disease, are best imaged by MRI. MRI assists in the early detection of rheumatoid arthritis, which allows earlier initiation of treatment. PMID- 15828370 TI - [Usefulness of anti-citrullinated protein antibodies in the diagnosis and prognosis of rheumatoid arthritis]. AB - Rheumatoid factors were the first biological markers used in diagnosis of rheumatoid arthritis. Their detection is rather sensitive but lack specificity. In contrast, anti-citrullinated protein antibody detection has recently been shown to have a reasonable sensitivity and a high diagnostic specificity. These autoantibodies can be present early in the disease and predict erosive arthritis. Even if an improvement of sensitivity is desirable, the anti-citrullinated protein antibody detection meets many of the requirements of a useful serological marker for rheumatoid arthritis. Its prognostic potential may provide an indication for aggressive treatment. Consequently, incorporation of anti citrullinated protein antibody positivity as an extra criteria would improve the accuracy of the American College of Rheumatology criteria, commonly used for disease classification. PMID- 15828371 TI - [Treatment of rheumatoid arthritis in 2005: prompt, agressive and customized]. AB - Treatment of rheumatoid arthritis in 2005: prompt, aggressive and customized Rheumatoid arthritis can be extremely serious (joint destruction, functional loss, decrease in life expectancy). Fortunately, our therapeutic means have recently progressed enormously (better appreciation of efficacy and ways to use DMARDs combinations and new molecules such as leflunomide and anti-TNFs, understanding of the importance of early adequate and intensive treatments when necessary). Huge progresses have also been performed with regards to evaluation and follow-up strategies (disease activity score--DAS, health assessment questionnaire--HAQ), which allows us to adapt the treatment much better. The goal now can and must be quick and total remission of the disease in all patients thus avoiding as much as possible irreversible joint damages with accompanying morbidities. PMID- 15828372 TI - [Rheumatoid arthritis and pregnancy]. AB - Rheumatoid arthritis occurs frequently in women in childbearing years. With the improvement of the treatments, more patients with rheumatoid arthritis consider a pregnancy. Close co-operation between the physician and the obstetrician caring for the mother and the foetus is necessary. The disease should be well controlled at the time of the conception, although an amelioration of rheumatoid arthritis occurs in about 75% of pregnancies, in the first trimester. Some medications can be used during pregnancy and lactation. There is no indication of any adverse effects of rheumatoid arthritis on pregnancy outcome. The mother needs to be followed up regularly after delivery because of the high risk of post-partum flare. PMID- 15828373 TI - [Rheumatological arthroscopy]. AB - Arthroscopy is a useful tool for the clinical rheumatologist. It allows the clinician to examine the synovial tissue under direct vision for evidence of active inflammation. It permits tissue sampling. Biopsies taken at arthroscopy can be used for diagnosis, as in the case of infectious arthritis, and in the context of clinical research. Arthroscopy also has therapeutic benefits when performed in conjunction with lavage. It is a simple procedure to perform, after the appropriate training. The procedure can be carried out on an outpatient basis and is well tolerated by patients. In this article we will outline the practical concerns relating to rheumatological arthroscopy. PMID- 15828375 TI - [Intestinal stoma: preoperative and postoperative management]. AB - The aim of this review is to present the management and indications of intestinal stomas. A stoma induces a body image alteration with important familial and social consequences. A preoperative visit to the stoma nurse prevents technical and/or psychological complications. Stoma nurses, surgeons and general practionners work together to help the patient in his/her new life. New stoma devices have also contributed to improve quality of life. Social and sexual activity can be maintain despite intestinal stoma with appropriate education. PMID- 15828376 TI - [Return from Argentina: a clandestine cutaneous passenger]. PMID- 15828377 TI - [Back to Francois Mitterrand, the lie and the medical secret]. PMID- 15828378 TI - [Eliminating the contract duty, at best, will not eliminate any problem!]. PMID- 15828379 TI - [A new osteo-articular imaging]. PMID- 15828380 TI - [Cloning: it is just a beginning]. PMID- 15828381 TI - [New in Russian healthcare legislation]. PMID- 15828382 TI - [The specificity of hygienic behavior of virtually healthy population]. AB - An analysis of the results of a sociological study showed that both men and women aged around 50 and noting subjectively no chronic pathology regard health as a life value. Social hardships reliably worsen the preventive activity and limit its variability. An essential share of population (above 30%), while taking care of its health, does not give up such harmful habits as smoking and abuse of alcohol. The described data denote the need in not so much propagating the healthy mode of life as in motivating it. The disturbing trend towards ignoring the motivations targeted at health promotion deserves special attention; the above trend is observed in absence of age-related chronic pathologies and against the background of an improving social status especially in men. PMID- 15828383 TI - [The life quality of population belonging to senior age groups]. AB - Described in the paper are issues related with the involution time in men and women, who are in the second period of their mature age, and with complications of the mentioned time interval affecting the life quality and its expectancy and aggravating the risk of sudden death. It was for the first time that the authors undertook a complex study of men and women of the mentioned age whose involution was physiological or complicated; they focused attention on the criterion specificity of the involution disregulations of different nature affecting the mean life expectancy and life quality in patients. The study results verify to the fact that the involution period is essentially influenced not only by endogenous but also by exogenous factors. It is pointed out as necessary to promote the primary prevention and outpatient follow-up of men and women aged above 40 for the purpose of exposing the factors of risk of involution complications and for the purpose of preventing early complications observed during the discussed life period. It is demonstrated that this would essentially improve the life quality in a significant share of the nation including patients aged above 40. PMID- 15828384 TI - [Health of population in Arkhangelsk Region]. AB - Presented in the paper are data on the health of population in Arkhangelsk Region. Specific features of morbidity are defined. Parameters of morbidity are described; its structure is evaluated and retrospectively analyzed. The key trends are displayed for the recent 5 years, they are: an increasing number of acute diseases, many of which become chronic; and growing primary disability. The regional specificity of morbidity must be taken account of in structuring of medical care. PMID- 15828385 TI - [Risk factors in hypertension]. AB - The authors studied the medical-and-social factors of risk of the hypertension disease in the family environment. The below main factors of medical-and-social risk of the disease were respectively denoted for husbands and wives: men- medical history compromised by ischemic heart disease, continued psycho-emotional tension and by non-observance of the modes of meals, labor and recreation; women- continued psycho-emotional tension, malnutrition, failure to comply with medical prescriptions and smoking. PMID- 15828386 TI - [Inflammatory diseases in male teenagers]. PMID- 15828387 TI - [Monitoring of anthropologic parameters and of the functional condition in enrollees]. AB - There is an analysis of a monitoring of health parameters in enrollees of a social university. The results confirm the hypothesis on that the health of schoolchildren and students has been changing for the worse. It is proven as necessary to set up, at high schools, systems of health management applicable to students, including health-improving measures for male students, whose health has been declining, according to some parameters, faster than in female students. PMID- 15828388 TI - [Prevention of iodine-deficient diseases]. AB - The Interregional center of public health and development assisted by Sechenov Moscow Medical Academy undertook, 1999-2000, a study of the standard of knowledge, attitude and conduct of consumers in respect to the use of iodinated salt (IS) in the prevention of iodine-deficient diseases (IDD). It was demonstrated that, on the average per one district, 19% of city respondents and 13% of rural respondents used IS only. About 31% of city respondents used IS when it was available at the next-door shop, i.e. from time to time. This figure reached 48% in Irkutsk and Orenburg Regions. The share of those who used IS sometimes in rural regions made an average of 20.7%. 67.2% of those who believe that IDD can be prevented think that ID can also be prevented. Less than 5% of them say ID cannot be regarded as a reliable tool in the prevention of IDD. PMID- 15828389 TI - [Prevalence of smokers in Moscow Region]. AB - The results of monitoring of bad habits (smoking) among senior schoolchildren in 23 towns of Moscow Region are dynamically described in the paper for 2000-2003. Smoking-related specific features are analyzed with respect to the age and sex differences of teenagers. A database, comprising above 25,000 entries, was created on the prevalence of smokers among city schoolchildren. PMID- 15828390 TI - [Monitoring of congenital defects in children of Nizhniy Novgorod Region]. AB - Presented in the paper are the results of monitoring of congenital defects (CD) among newborns in Nizhniy Novgorod Region in 1999-2003. All CD were subject to routine registration at maternity homes, pediatric clinics and dissection rooms of hospitals. The mean CD prevalence made, during the mentioned period, 15 cases per 1000 newborns, which is in keeping with the CD figures registered in other regions of Russia. The share of some nosological CD forms is in line with the results of the International Registry. PMID- 15828392 TI - [Dental aspects of health of children and teenagers in Kursk Region]. PMID- 15828391 TI - [Infant mortality in Leningrad Region]. AB - Presented in the paper is the level and dynamic figures (for a period of 15 years) of infant mortality, i.e. one of the most important indices of population health and of quality of medical care, including all its components, as well as of perinatal mortality as observed in Leningrad Region. The main causes of infant mortality are described.; the key measures on its prevention are defined. PMID- 15828393 TI - [Reforming of the primary medical and sanitary care in a highly urbanized region]. PMID- 15828394 TI - [An up-to-date model of organization of pediatric psychiatric care in a big city]. PMID- 15828395 TI - [Importance of pharmacological and economic research of rational use of drugs]. PMID- 15828396 TI - [Quality and efficiency of labor of nurses at city clinics]. AB - The case study was based on 4 clinics of the city of Makhachkala, Republic of Dagestan. Three hundred and fifty-five nurses and 352 patients were questioned as to the organization of labor of nurses at city clinics. The main activities of 55 nurses were subject to an expert evaluation. PMID- 15828397 TI - [Transfer of a child from children's home to family: medical aspects]. AB - The project "Leaving of babies at children's homes and deinstitutionalization" was implemented at the children's home of Tbilisi. The purpose was to find out what influence was produced by deinstitutionalization on the physical and psychomotor development of deprived babies. The study lasted for 12 months. 3 groups of children were investigated. Group 1 comprised children selected for deinstitutionalization, age-matching healthy children who stayed at the children's home were in group 2, and healthy children randomly selected from 2 nurseries of Tbilisi were in group 3. The parameters of anthropometry and psychomotor development were evaluated according to the Denver screening test at the very beginning and in 12 months. The children from the children's home almost caught up, in 12 months after deinstitutionalization, with their control matches. Whereas, the children, who were brought up at the children's home, were behind the group-1 children and controls by all parameters. Projects promoting the upbringing of deprived children in their own or foster families are recommended for implementation. PMID- 15828398 TI - [Professional rehabilitation of the disabled in the Nizhegorodski region]. AB - Questioning of one hundred and eighty disabled persons of the able-bodied age denoted high levels of education and economic activity among them. However, the state guarantees of professional rehabilitation of the disabled are not up to the mark. Suggestions are made of how to promote the professional rehabilitation of the disabled of the able-bodied age. PMID- 15828399 TI - [Financing of regional healthcare in Kazakhstan]. AB - It is suggested in the paper how to finance stages of healthcare in Kazakhstan. The introduction of the Budget Code is expected to change the functioning of the financial department for the better. PMID- 15828400 TI - [Military medicine mirrored by press]. PMID- 15828401 TI - [Emigration of Russian doctors to Bulgaria]. PMID- 15828402 TI - [I. F. Fitingof--a reformer of state medical education in Russia]. PMID- 15828403 TI - Community-building principles: implications for professional development. AB - This article reviews a Think Tank meeting among child welfare practitioners at the 2003 Building Communities for 21st-Century Child Welfare Symposium. The Child Welfare League of America's focus on community building is recognition of the vital importance of promoting and fostering collaboration with community members to enhance the well-being of children, families, and communities. The Think Tank participants responded to four questions concerning the knowledge, policies, and strategies that are needed for the development of strategies for community building and child welfare. This article highlights several of the findings of the preconference, which addressed the challenges and opportunities inherent in community-building practices and discusses the key principles that emerged from the Think Tank. The article emphasizes implications for professional education and cites selected examples of innovative community-building initiatives with families. PMID- 15828404 TI - Moving mountains together: strategic community leadership and systems change. AB - Traditional leadership practice frameworks to guide systems change often fall short in today's practice environment. Reclaiming Futures is a national initiative to create an integrated, comprehensive, seamless system of care for teens with substance abuse problems involved in juvenile justice. It uses leadership and systems reform strategies to improve public health institutions. The premise is that interruption of the destructive cycle of drugs and delinquency can only be attained through the cultivation of shared strategic leadership. This article presents the limitations of traditional practice frameworks of leadership, describes the theory of this new approach to leadership development, and clarifies collaborative leadership and system change. The article includes an example of the framework in action, a discussion of lessons learned, and recommendations for community leadership development initiatives. PMID- 15828405 TI - The New York City neighborhood-based services strategy. AB - The New York City Administration for Children's Services (ACS) instituted a neighborhood-based services system through the realignment of all foster care, preventive, and protective services along community district lines. ACS, with its community partners, also formed neighborhood-based networks to improve service coordination and collaboration among key community stakeholders and to shape a multisystem strategy tailored to each district informed by child welfare data. Based on analysis of neighborhood-specific census tract child welfare data, ACS initiated the Community Partnership to Strengthen Families project to address the disproportionate number of foster care placements originating from a small group of high-need communities, including Manhattan's Central Harlem. This article describes examples of specific strategies based on the Central Harlem experience. PMID- 15828406 TI - "Nothing about me without me": leading the way to collaborative relationships with families. AB - This article discusses the National Center on Addiction and Substance Abuse's CASA Safe Haven, an evidence-based, community-driven intervention program for children and families in child welfare whose lives have been adversely affected by substance abuse, and for staff in the agencies that work with them. CASA Safe Haven builds collaborative relationships that feature a blend of multidisciplinary teams that share responsibility for helping families; family group conferencing, in which families are equal and welcome participants in designing and driving a service plan; and the influence of family court to hold families and service providers accountable for progress. CASA Safe Haven is a framework for collaboration. PMID- 15828407 TI - Drug-endangered children need a collaborative community response. AB - The United States is facing an epidemic of the use of methamphetamine drugs. Child welfare has not yet addressed the needs of the children living in so-called "meth homes." These children are endangered not only from the chemicals involved, but also from parental abuse and neglect. Communities are recognizing the need for interagency collaboration to address the consequences of this epidemic. Spokane, Washington, has created a Drug-Endangered Children Project, whose mission is to implement a collaborative response among law enforcement, prosecutorial, medical, and social service professionals to the needs of drug endangered children. This article presents the findings from the evaluation of the first year of the project, including a baseline assessment of the needs of drug-endangered children and the extent of community-based collaboration achieved. This article makes recommendations for future community-based partnerships to improve the well-being of drug-endangered children. PMID- 15828408 TI - Seeds of change: using peacemaking circles to build a village for every child. AB - Roca, Inc., a grassroots human development and community organization, has adopted the peacemaking circle as a tool in its relationship building with youth, communities, and formal systems. Circles are a method of communication derived from aboriginal and native traditions. In Massachusetts, the Department of Social Services and the Department of Youth Services are exploring the application of the circle in programming with youth and families. By providing a consistent structure for open, democratic communication, peacemaking circles enhance the formation of positive relationships in families, communities, and systems. The outcome is a stronger community with greater unity across truly diverse participants. This article presents the theory and practice of peacemaking circles, the lessons and challenges of implementing circles in formal organizations, and the potential of the circle to support a strengths-based and community-based approach to child welfare. PMID- 15828409 TI - Co-production dynamics and time dollar programs in community-based child welfare initiatives for hard-to-serve youth and families. AB - Hard-to-serve youth and families residing in high-poverty communities often have multiple, interlocking needs. These needs necessitate complex service models. The complex model described in this article combines a unique approach to wraparound services with a coproduction framework and related theories. The model aims to improve outcomes for vulnerable youth and their families, simultaneously strengthening communities by employing residents and engaging participants in community service. Examples derived from current pilot projects illustrate co production's importance for other child welfare initiatives. PMID- 15828410 TI - Unleashing the power of community to strengthen the well-being of children, youth, and families: an asset-building approach. AB - Search Institute's decade-plus emphasis on the elements of positive human development and community approaches to asset building can make a meaningful contribution to the field of child welfare. The institute's framework of developmental assets identifies a set of interrelated experiences, relationships, skills, and values that are associated with reduced high-risk behaviors and increased thriving behaviors. Its community-building work emphasizes the human relations and developmental infrastructure children, youth, and families require for their health and well-being. PMID- 15828411 TI - Understanding communities today: using matching needs and services to assess community needs and design community-based services. AB - Matching Needs and Services (MNS) is a practice tool intended to help people who work with vulnerable children use rigorously assembled information on needs as a guide to design, implement, and evaluate more-effective services. To do this, MNS focuses on needs but links them to outcomes and thresholds before dealing with the services to achieve those outcomes. PMID- 15828412 TI - "Just therapy" with families on low incomes. AB - This article addresses the inadequacies of counseling, therapy, and social work that occurs with low-income families. The author argues that many families who seek help arrive with problems that are usually assessed separately from their socioeconomic and cultural contexts. Careful questioning will often lead to the discovery that the onset of many family problems are located in events external to the family, such as unemployment, bad housing, and racist, sexist, or heterosexist experiences. They can be extremely depressing ongoing experiences that eventually lead parents and children into a state of stress that opens them up to physical and mental illnesses. This article argues that a wide body of research supports such a view and that counseling, therapeutic, and social work practices should address these issues much more directly. It also argues that practitioners have an important role to play in social and economic policy development out of respect for their clients' struggles. PMID- 15828413 TI - Understanding communities of neglectful parents: child caregiving networks and child neglect. AB - This article focuses on family social networks and the community of caregivers of neglected children. If neglect is part of family functioning, who watches over the children? Using a case study approach, this study researched 12 children and their parents. Several concepts, such as multiple caregiving and kin keepers, revealed that study children were cared for by many people. Social network mapping used in this study indicated that families were not isolated from the larger community, had various forms of negative and positive social support, were low income, and were involved in substance abuse and domestic violence. Understanding the patterns that emerge from the complex web of family, friends, social service agencies, and the larger social community in which neglected children live can result in better community building. PMID- 15828414 TI - All it takes is leadership. AB - The authors, as leaders in a public child welfare system, have teamed together and reached out to their private sector partners in a large, rural county in western Colorado. This effort was part of a comprehensive, communitywide effort to redesign and fundamentally improve the entire child welfare service delivery system. Across the country in many areas where collaboration and integration have been the focus, we often hear voices in the private and public sector declaring the importance of integration. Why, then, does it not happen as a general course of action? The authors believe the answer lies in leadership, both public and private. They hold the Mesa County model up as witness to that fact. PMID- 15828415 TI - From tragedy to triumph: a segue to community building for children and families. AB - In 2000, more than 60 nonprofit agencies, health care providers, government officials, and community advocates in Tarrant County, Texas, came together to work for systemic change in the mental health care system. The coalition, known as the Mental Health Connection, began working toward a "No Wrong Door" approach to mental health services, which required aggressive coordination between federal, private, and nonprofit resources. The result is a five- to six-year plan for implementation of a new systems of care model for children with severe emotional disturbances and their families. The Mental Health Connection also focuses on legislative advocacy to bring about necessary policy changes at the local, state, and federal levels. Finally, the coalition focuses on developing sustainable revenue streams that will allow the new systems to remain in place once the group accomplishes the initial mission of the Mental Health Connection. PMID- 15828416 TI - Rebuilding Native American communities. AB - The Wellbriety Movement in Native American communities draws on the wisdom and participation of traditional elders. Beginning with a basic community teaching called the Four Laws of Change and the Healing Forest Model, the Wellbriety Movement blends Medicine Wheel knowledge with the 12 Steps of Alcoholics Anonymous to provide culture-specific addiction recovery help for Native Americans. The four Journeys of the Sacred Hoop brought the Wellbriety Movement to Native and non-Native people alike from 1999 to 2003. PMID- 15828417 TI - [I.S. Beritashvili (Beritov) is one of the founders of the modern biobehavioral science (to the 120th anniversary)]. AB - Ivan Solomonovich Beritashvili (Beritov) is one of the great Russian physiologists who have created the Russian route of the modern science of behavior. He has proposed and experimentally substantiated a concept of psychoneural activity according to which the behavior controlled by images is the main and higher form of the behavior of vertebrates. Behaviors on the basis of conditioned reflexes and images are qualitatively different activities underlain by different neural substrates. Recently, the Beritashvili's principle ideas have been confirmed by multiple experimental studies of the animal and human cognitive behavior. At present, the concept of the procedural memory formed by training and acquisition and episodic memory formed on the basis of images is commonly accepted. The episodic memory stores the environmental events and phenomena perceived by an animal or a human. Each time when the image is reproduced during perception of environment or its part, an animal performs the same behavioral act as during the actual perception. Beritashvili's viewpoint that the procedural and declarative memories are controlled by different brain structures has been confirmed by experiments. PMID- 15828418 TI - [Characteristics of the heart rate during mental tracing of heartbeats in healthy subjects]. AB - Mental counting of self heartbeats results in changes of parameters of heart rhythm--domain and other frequencies of the RR-intervals, especially, high frequencies (0.15-0.4 Hz), which are relevant to respiration. The accuracy of mental counting of the heartbeats (the ratio of traced beats to the number of registered ones) corresponds to individual relations between the frequencies, regularity, and total number of the heartbeats. A possibility of the voluntary regulation of respiration as the cause of changes in the heartbeat frequency is discussed in terms of the center-periphery integration (system approach) for optimal perception of self heartbeats. PMID- 15828419 TI - [Changes in components of the auditory long-latency evoked potentials at different stages of the slow-wave sleep]. AB - In accordance with the present views, during sleep, analysis of external stimuli continues at the subconscious level, because the need to estimate the biological significance of external stimuli in order to maintain a flexible contact of a sleeping subject with the environment persists during sleep. It is known that new components of the auditory evoked potentials (AEP) appear as sleep deepens. However, the common procedure of analysis of event-related potentials averaged for a group of subjects has some drawbacks because of the interindividual variability of the event-related potentials. Therefore, an additional analysis of the interindividual variability of the AEP shape and component structure can simplify the detection of individual components of group-averaged AEP at different stages of the slow-wave sleep. The AEPs were recorded in healthy volunteers (n = 26) during falling asleep in the evening from eight EEG derivations (F3, F4, C3, C4, P3, P4, O1, O2) in reference to a linked mastoid electrode. Computer-generated sound stimuli (50 ms-pulses with the frequency of 1000 Hz, 60 dB HL) were presented binaurally through earphones with interstimulus intervals of 20-40 s. Selective summation of AEPs for all the subjects was performed for each stage of the slow-wave sleep individually for each of the eight derivations. It was shown that the account made for interindividual variability of the AEP shape facilitated the identification of individual components of the group-averaged AEP typical of wakefulness (P1, N1, P300) and those which appeared during sleep onset and at different stages of the slow-wave sleep (P2, N350, P450, N550, N900). PMID- 15828420 TI - [Choice between probability and value of alimentary reinforcement as means for revealing individual typological features of dog behavior]. AB - Individual typological features of behavior of dogs were investigated by the method of choice between the low-valuable food available constantly and food of high quality presented with low probability. Animals were subjected to instrumental conditioning with the same conditioned stimuli but different types of reinforcement. Depression of a white pedal was always reinforced with meat bread-crumb mixture, depression of a black pedal was reinforced with two pieces of liver (with probabilities of 100, 40, 33, 20, or 0%). The choice of reinforcement depended on probability of valuable food and individual typological features of the nervous system of a dog. Decreasing the probability of the reinforcement value to 40-20% revealed differences in behavior of dogs. Dogs of the first group, presumably with the weak type of the nervous system, more frequently pressed the white pedal (always reinforced) than the black pedal thus "avoiding a situation of risk" to receive an empty cup. They displayed symptoms of neurosis: whimper, refusals of food or of the choice of reinforcement, and obtrusive movements. Dogs of the second group, presumably with the strong type of the nervous system, more frequently pressed the black pedal (more valuable food) for the low-probability reward until they obtained the valuable food. They did not show neurosis symptoms and were not afraid of "situation of risk". A decrease in probability of the valuable reinforcement increased a percentage of long latency depressions of pedals. It can be probably suggested that this phenomenon was associated with increasing involvement of cognitive processes, when contributions of the assessments of probability and value of the reinforcement to decision making became approximately equal. Choice between the probability and value of alimentary reinforcement is a good method for revealing individual typological features of dogs. PMID- 15828421 TI - [Correlation of unit activity of the right and left amygdala in food motivation and emotional stress]. AB - Correlation between activities of neurons in the right and left central nuclei of amygdala of rabbits recorded during quiet wakefulness, after 24-h food deprivation, after satiation and during emotional stress (demonstration of a dog) was studied by plotting crosscorrelation histograms. The histogram peaks shifted from zero were observed in 50-67% cases. In hungry animals, in a greater number of cases (52%), the discharge of a neuron in the left amygdala was the first in a pair, and the discharge of the right neuron was delayed (peaks from 10 to 50 and from 130 to 150 ms). The opposite order of discharges was less frequent (36%). When a rabbit saw a dog, the number of common inputs to neurons increased and the leading role of the right amygdalar neurons grew (57%) due to an increase in inhibitory influences from the right to the left amygdala. In most cases, the interaction between amygdalar neurons occurred at the frequencies of the delta range, predominantly, from 2 to 4 Hz. PMID- 15828422 TI - [Spontaneous activity of hippocampal neurons and stimulation rhythm acquisition in young rabbits during learning: age features]. AB - Ontogenetic mechanisms of memory formation were studied using an experimental model of conditioned reflex to time, i.e., trace acquisition of a stimulation rhythm by hippocampal CA1 neurons of young (1-4 weeks old) and adult rabbits (5-6 months old). It was found that age-related development of learning ability includes several stages: complete absence of memory traces (6-7 days old), rapid acquisition without consolidation (8-14 days old), and formation of perfect memory (25-30 days old). Both specific and nonspecific changes in spontaneous activity of neurons were observed. Changes in the rate of discharges related to rhythmic stimulation were accompanied by changes in spontaneous activity. With the development of an animal, spike activity increased in parallel with improving of the functional properties of neurons, their structural organization, formation of the afferent contacts in the hippocampus completed after a period of three weeks from birth, and formation of metabolic processes, modulatory systems, and traffic function of hippocampal neurons. A capability for plastic reorganization is of great importance for adaptation mechanisms and conditioned behavior of a developing animal in accordance with structural maturation and development of the functional regulation of neuronal reactivity in the hippocampus. PMID- 15828423 TI - [Computation of color and brightness differences by neurons in the rabbit visual cortex]. AB - Changes in activity of 54 neurons in the rabbit visual cortex evoked by the replacement of eight color and eight achromatic stimuli in pairs were analyzed. The diffused stimuli generated by color SVGA monitor were used in the experiments. The earliest response of phasic neurons (50-90 ms after the replacement) was strongly correlated with differences between stimuli in color or intensity. This response ("the signal of differences") was used as a basis of a matrix (8 x 8) constructed for each neuron. Such matrices included mean numbers of spikes per second in responses to changes of different stimuli pairs. All matrices were subjected to factor analysis, and the basic axes (the main factors) of sensory spaces were revealed. It was found that 16 neurons (30%) detected only achromatic differences between stimuli. Perceptual spaces of these neurons were two-dimensional with brightness and darkness orthogonal axes. The spaces of 12 neurons (22%) were four-dimensional with two chromatic and two achromatic axes. The structure of the perceptual space reconstructed from neuronal spikes was similar to the space calculated from the early VEP components recorded under similar conditions and to another space reconstructed on the basis of rabbit's instrumental learning. The fundamental coincidence of color spaces revealed by different methods may reflect the general principle of vector coding in the visual system and suggests the coexistence of two independent cortical mechanisms of the detection of chromatic and achromatic differences. PMID- 15828424 TI - [Glutamate release in the nucleus accumbens during competitive presentation of aversive and appetitive stimuli]. AB - By means of in vivo microdialysis combined with HPLC analysis, we have shown that extracellular glutamate level in the rat n. accumbens increases during a simultaneous presentation of a palatable diet and a tone previously paired with a footshock, the magnitude of the extracellular glutamate increase being proportional to the latency of food taking. In contrast, extracellular glutamate level remains unchanged when the diet is presented after the conditioned aversive stimulus or when the tone is given alone. These data suggest that the glutamate release evoked by the competitive presentation of the diet and the conditioned aversive stimulus appears to be related to the inhibition of a planned feeding response, whereas the choice between behavioural strategies may not contribute to this phenomenon. PMID- 15828425 TI - [Influence of chronic melipramine administration abolition on locomotion and defensive conditioned reflexes in passive and active avoidance in rats]. AB - The chronic (21 days duration) administration of tricyclic antidepressant melipramine of Wistar rats strain (15 mg/kg daily, intraperitoneally) evoked weight loss of animals. The 7 days after melipramine abolition its sedative effect was observed in the "open field" test by decrease of locomotion and the number of boles. The 7 and 14 days after melipramine abolition the difference between control and melipramine treated animals in passive and active avoidance learning and memory not found. The experimental results comparison with the literature data show, that chronic melipramine administration of intact animals evokes a sedative state. This conclusion does not contradict to idea of punishment function of brain serotoninergic system. PMID- 15828426 TI - [The effects of artificial analogue peptide TSKY isolated from the brain of hibernating ground squirrels in rats and mice]. AB - Elevation of the i.c.v. injection dose of TSKY from 4 to 8 microg increased the movement activity of rats; in EEG theta- and beta-rhythms were enhanced and alpha rhythm was suppressed. On the contrary, after treatment of 15 microg the rats fell into sleepy-like state; theta- and beta2-rhythms suppression, delta-, alpha- and beta1-rhythms were increased. Exposure under hypoxia-hypercapnia conditions reduced body temperature of mice to 18-19 degrees C, and maintain this state about 3-4 h after transferring into conventional gas medium. Preliminary cooling mice were administrated with TSKY that at dose 100 microg intraperitonally induced a prolonged hypothermia up to 12 h. Analogous injection without cooling raised mice temperature by 1.2 degrees C during about 2 h. PMID- 15828427 TI - [Functional role of neuropeptide CNP4 encoded by gene HCS2 in Helix lucorum nervous system]. AB - Helix Command Specific 2 (HCS2) gene is constantly expressed in parietal premotor (command) interneurons involved in control of the terrestrial snail Helix lucorum withdrawal behavior as a trigger element. It is also expressed under noxious conditions in other neurons presumably involved in withdrawal behavior. In this study we addressed the role of neuropeptide CNP4, encoded by gene HCS2, in the regulation of activity of the respiratory system, and in the influence on growth of isolated neurons in culture. It was shown that activity of the premotor interneuron elicits a direct effect (pneumostome closure), and a delayed intensification of respiratory movements. Application of CNP4 mimicked the delayed effects. Presence of the peptide CNP4 in solution for cultured neurons led to increase of neuronal growth. Immunochemical localization of the protein precursor encoded by gene HCS2 and peptide CNP4 in the cultured premotor interneurons revealed their preferential presence in the growth cones. The obtained results suggest that CNP4 may be secreted and involved in synergic regulation of behavior of a snail. PMID- 15828428 TI - [The role of opioid system in the regulation of the sleep-wakefulness cycle]. AB - In chronic experiments influence of single administration of various doses of morphine (0.5, 2.0 and 3.0 mg/kg) on general behavior, structural organization of the sleep-waking cycle and some EEG and somato-vegetative parameters was studied in the cats. Intraperitoneal administration of morphine elicits numerous autonomic disorders, psychic derangement and complete deterioration of the sleep waking cycle structure. The result certify that single administration of morphine induces activation of the opiate receptors in various structures of the brain, which entails deviations of behavioral and EEG parameters, activation of the structures responsible for initiation of behavioral waking, suppression of activity in those structure, which control the slow wave sleep triggering mechanisms, deterioration of the sleep quality and significant elevation of emotional tension against the baseline. On the basis of obtained data it is suggested that the opioid system plays an important role in the sleep-waking cycle regulatory mechanisms and other forms of brain integrative activity in general. PMID- 15828429 TI - [Neuronal septal activity during hippocampal seizure discharges generation in acute model of epilepsy]. AB - The activity of the neurones of the medial septal region (MS) and the hippocampal EEG in control and during the appearance of seizure discharges provoked by electrical stimulation of the perforant path were investigated in the awake rabbit. During afterdischarge generation in the hippocampus the dense neuronal bursts separated by periods of inhibition were recorded in the MS. In one group of neurons the bursts of spikes coincided with the discharges in the hippocampus, in other group-occured during inhibitory periods. When the afterdischarge stopped, in the septal neurons with theta activity the disruption of theta pattern was recorded, which have been correlated with the occurrence of low amplitude high frequency (20-25 Hz) waves in the hippocampal EEG. As a rule, the neuronal activivity of the MS recovered much quickly than EEG of the hippocampus; in some cases the increasing of the theta regularity was observed. The definite accordance of the electrical activity of the hippocampus and MS during seizure discharges suggests that the septohippocampal system operate as integral nervous circuit in these conditions. Diverse in the temporal interrelations between the discharges of MS neurones and ictal discharges in the hippocampus in the different cells possible indicate that various groups of the septal nervous elements have different participation in the seizure development. Appearance of the high frequency bursts in the MS is a possible "precursor" of the seizure onsets. PMID- 15828430 TI - [Remote effects of early postnatal pituitary hormone melatonin injection on audiogenic seizures in Krushinsky-Molodkina rats]. AB - Rats Krushinsky-Molodkina inbred strain (KM) genetically prone to audiogenic seizures were injected with pineal hormone melatonin (50 mg/kg, s.c.) within the period 7th to the 14th posnatal days (PND). The remote effects of this injection adult KM rats consisted in a decrease in the latency and increase in severity of myoclonic seizures produced by audiogenic kindling (20 sound stimuli, 100 dB and 12-15 kHz). As compared to the control, in the cortex and hippocampus of rats of melatonin group, we also found a significant reduction of both total and functional activity of Ca2+/calmodulin-dependent protein kinase II (CAMK II) after audiogenic kindling. On the contrary, melatonin administration within the 1st to 7th PND and the 14th to the 21st PND resulted in a decrease in seizure activity. In the first case, both the total (cortical) and functional (hippocampal) CAMK II activities in melatonin-injected rats were increased as compared to control, whereas in the second case, only a slight increase in Ca2+ independent CAMK II activity in the hippocampus of melatonin-injected rats was observed. Probably, the melatonin administration in the period of early postnatal development changes the features of expression and/or regulation of CAMK II activity, and this could be one of the mechanisms of audiogenic seizure modulation in KM rats. PMID- 15828431 TI - [Digital registration and analysis of visual information in behavioral experiment]. AB - A new software for recording, registration and computer analysis of behavior of laboratory animals, EthoStudio, has been developed. The software allows the latency, number and duration of up to 10 behavioral patterns to be simultaneously registered with parallel recording of video on a hard disk. The video records can be played back to track animal locomotion using a computer. EthoStudio was applied to study mouse behavior in the open field test. The numbers of rearings and groomings as well as accumulated time of grooming were interactively assayed by pressing and holding buttons on a computer keyboard. The number of squares crossed, the number of entries into the center and the time spend therein were assayed by computer analysis. EthoStudio can be applied to objectively study various kinds of animal behavior with reliability and accuracy necessary for modern ethological, physiological and pharmacological experiment. PMID- 15828432 TI - The Rockefeller Foundation and the prevention of malaria in Corsica, 1923-1951: support given to the French parasitologist Emile Brumpt. AB - The role of the International Health Board of the Rockefeller Foundation in campaigns for malaria control in many countries has clearly been documented extensively. In contrast, the involvement of the Rockefeller Foundation in the control of malaria in France has not been reported yet. The present paper describes the way in which the Rockefeller Foundation got involved, along with the parasitologist Emile Brumpt, in the anti-malaria campaigns in Corsica, France, between 1924 and 1951. We analyze the scientific and technological strategies used for that purpose and the manner in which the Rockefeller Foundation policy had influenced Brumpt's actions. The unusually long support of the Rockefeller Foundation to Emile Brumpt (1924-1948) and to research and teaching Parasitology in the Faculte de Medecine de Paris is also discussed. PMID- 15828433 TI - [Two new human cases of dirofilariasis in Sardinia]. AB - Two new human cases of dirofilariasis, both due to Dirofilaria (Nochtiella) repens, recently occurring in Nuoro province (Sardinia) on people permanently resident in the island, are described. The parasite was located in one case into the uterine round ligament, in the other it was subconjunctival. Some observations about the diagnostic difficulties in the first one and the dramatic symptoms in the second, whose parasite was yet alive during the surgical ablaction, are discussed. These cases are the 9th and the 10th, respectively, reported in man in Sardinia, but it is possible that other cases are undiagnosed and consequently are not published. PMID- 15828434 TI - Sequencing of the complete gene coding for the GroEL of the Wolbachia of Dirofilaria immitis and expression and purification of the recombinant protein. AB - Wolbachia are intracellular bacteria that infect arthropods and filarial nematodes. These bacteria play an important role in the immunology and pathogenesis of filarial diseases through their proteins and, possibly, other molecules. GroEL is a constitutively expressed bacterial protein; it is highly conserved among bacteria and is involved in the correct folding of newly synthesized proteins. Here we report the production of recombinant GroEL from the Wolbachia of Dirofilaria immitis. Our goal is to test the hypothesis that GroEL is involved in the immunopathology of filariases. The complete groel gene was PCR amplified, sequenced and cloned into an expression vector. The recombinant GroEL was purified by affinity chromatography by using high-performance liquid chromatography (HPLC). PMID- 15828435 TI - Necropsy and coprology in wild boar (Sus scrofa) in Livorno Mountain Park (Tuscany, Central Italy). AB - The present research analyses the reliability of coprological tests, both quantitative and qualitative, as indicators of the parasite burden of hosts, using data from wild boars (Sus scrofa) living in Livorno Mountain Park (Tuscany, Central Italy). In the case of intestinal strongyles, which turned out to be the dominant helminths of wild boars, the qualitative coprological test appears as a bad predictor of the real parasite situation of the herds, due to the high number of false negative results (34 animals out of 68). On the other hand, the positive predictive value of the test is high (90%). The quantitative test is significantly correlated with the individual parasite burden of wild boars. PMID- 15828436 TI - Ocular parastrongyliasis (=angiostrongyliasis): probable first report of human infection from a patient in Ceylon (Sri Lanka). AB - What is considered to be the first case of human parastrongyliasis (angiostrongyliasis) worldwide is described from a patient in Ceylon (Sri Lanka) in 1925. It also predates the description of the parasite in humans by Chen (1935). PMID- 15828437 TI - Some studies on three known and a new species of the genus Binema Travassos, 1925 (Travassosinematidae: Thelastomatoidea) from Manipur, North-East India. AB - During the investigation of insect nematodes of North-East India, a new species of Binema (Travassosinematidae: Thelastomatoidea) was recovered from the intestine of Gryllotalpa africana Beauv. It is described and illustrated as Binema anulinervus n.sp. Three known species namely B. ornata Travassos, 1925, B. mirzaia (Basir, 1942) Basir, 1956 and B. korsakowi (Sergiev, 1923) Basir, 1956 have also been collected, redescribed and illustrated in the present study. The new species differs from all known species and only comes close to B. pseudornatum Leibersperger, 1960 for having caudal flagellate appendage. Key to species of the genus and a comparative measurement chart are given in Table 1. PMID- 15828438 TI - [Italian state's quinine laws: the role of Giustino Fortunato and Angelo Celli. An introduction to the paper by Anna Celli "La lotta contro la malaria" reprinted form "Giustino Fortunato (1848-1932)," Ed. Archivio Storico per la Calabria e la Lucania, 1932. pp. 135-153]. PMID- 15828439 TI - In vitro activity, pharmacokinetics, clinical efficacy, safety and pharmacoeconomics of ceftriaxone compared with third and fourth generation cephalosporins: review. AB - Due to their wide spectrum of activity, good pharmacokinetics, established clinical efficacy and high tolerability, cephalosporins are among the most widely used antibiotics worldwide. The third and fourth generation cephalosporins are predominantly parenteral agents, administered two or three times daily, used in the treatment of a wide range of moderate to severe infections. Ceftriaxone, a third generation cephalosporin, is unique in exhibiting an unusually long elimination half-life that allows for once-daily administration. Among third generation cephalosporins, ceftazidime and cefoperazone are unusual among cephalosporins in possessing activity, albeit moderate, against Pseudomonas aeruginosa. However, both of these agents also exhibit marked loss of activity against Gram-negative organisms producing high levels of Class A or C beta lactamases. Sulperazone, a 1:1 combination of cefoperazone and the beta-lactamase inhibitor sulbactam, is more resistant to attack by Class A beta-lactamases but remains vulnerable to isolates producing Class C beta-lactamases. Ceftriaxone exhibits the widest antibacterial spectrum of third generation cephalosporins and this is reflected in clinical responses. Cefoperazone and sulperazone exhibit the poorest clinical responses. Although the fourth generation cephalosporins cefpirome and cefepime exhibit enhanced stability to bacterial beta-lactamases and marginally enhanced in vitro antibacterial activity over ceftriaxone, there is no clinical advantage in terms of clinical or bacteriological success. The cephalosporins are well tolerated, with few and generally transient adverse effects; the major exception being haematological abnormalities including blood coagulation disorders associated with cefoperazone. Several pharmacoeconomic studies indicate that the once-daily dosing regimen required for ceftriaxone is the major factor responsible for its cost-effectiveness over third and fourth generation cephalosporins. PMID- 15828440 TI - Antibiotic resistance in pathogenic Streptococcus pneumoniae isolates in Turkey. AB - Streptococcus pneumoniae causes various human infections such as meningitis, septicemia, otitis media, sinusitis, and pneumonia. Antibiotic resistance has already been reported with increasing frequency worldwide and is spreading. The earliest studies on pneumococcal antibiotic resistance go back to the late 1980s in Turkey. The resistance patterns have elevated with stepwise increments since then. By the beginning of 2005, approximately 40% of pneumococci were resistant to penicillin and nearly one-fifth of resistant isolates present high-level penicillin resistance. This proves that penicillin is still a good alternative for nonmeningeal infections. In addition, no ceftriaxone resistance have been reported in local Turkish studies, but cefuroxime, a second-generation cephalosporin, was recorded to have (10.8-20%) resistance rates. The most frequently assessed antibiotics other than penicillin in Turkish studies include erythromycin (4-19.4%), chloramphenicol (2-10%), clindamycin (2.5-13%) and tetracycline (13-28.6%) and all have various resistance profiles. On the other hand, nearly all or almost all of the isolates evaluated in Turkish studies are susceptible to rifampicin, quinolones, linezolid, quinupristin-dalfopristin and telithromycin. All these non-beta-lactam antibiotics except the tetracyclines are within acceptable limits of empirical approaches. Tetracycline must be used cautiously. In addition, trimethoprim sulfamethoxazole cannot be prescribed in probable pneumococcal infections since more than half of the isolates are nonsusceptible. PMID- 15828441 TI - Mechanisms of macrolide resistance in clinical pneumococcal isolates in a university hospital, Ankara, Turkey. AB - Macrolide resistance in Streptococcus pneumoniae is usually caused by the presence of the erm(B) or mef(A) resistance determinants. The aim of the present study was to identify the predominant macrolide resistance mechanisms among erythromycin-resistant S. pneumoniae isolated in a university hospital, Ankara, Turkey. A total of 669 S. pneumoniae strains were isolated from clinical specimens of patients admitted to the hospital between 1994--2002. The minimum inhibitory concentrations (MICs) of penicillin G, erythromycin A and clindamycin were determined by the agar dilution method according to NCCLS guidelines. Ninety one (13.6%) isolates were resistant to erythromycin. Erythromycin-resistant isolates were examined for their macrolide resistance phenotypes by a triple disc diffusion assay. It assigned 57 (62.6%) of the 91 erythromycin-resistant pneumococci to cMLS(B) phenotype, 19 (20.9%) to iMLS(B) phenotype and 15 (16.5%) to M phenotype. All erythromycin-resistant isolates were analyzed by PCR for the presence of erm(B) and mef(A) determinants. The isolates were characterized for the underlying resistance genotype, with 83.5% having erm(B), 16.5% having the mef(A) genotypes. This study provides further evidence of the dissemination of macrolide-resistant mutants in pneumococci as the use of new, long-acting macrolides increases. This is the first article about MLS(B) resistance phenotypes and genotypes of S. pneumoniae from Turkey and it emphasizes the need for future epidemiological monitoring of macrolide-resistant pneumococci. PMID- 15828442 TI - In vitro evaluation of faropenem activity against anaerobic bacteria. AB - Faropenem, a new oral penem with broad spectrum activity, could be used as empirical treatment in infections due to unidentified anaerobes, but only a few investigations have been carried out on these bacteria. The aim of this study was to compare faropenem in vitro activity with that of positive antimicrobial controls (metronidazole, imipenem, meropenem, amoxicillin, amoxicillin-clavulanic acid, ticarcillin-clavulanic acid, cefotetan, cefoxitin and clindamycin) against 462 anaerobic bacterial strains. The reference agar dilution method was used according to the NCCLS standard. Faropenem demonstrated high antimicrobial activity, similar to that of both imipenem and meropenem (faropenem Minimal Inhibitory Concentrations 50% and 90% were 0.12 and 1 mg/L for all Gram-negative anaerobes, 0.25 and 1 mg/L for all Gram-positive anaerobes). Only 5 strains of the Bacteroides fragilis group (1.1% of all anaerobes) were resistant to faropenem, which compared favorably with that of other reference antianaerobic drugs. The results obtained confirm those previously reported. PMID- 15828443 TI - In vitro synergy and selection of resistance by fluoroquinolones plus amikacin or beta-lactams against extended-spectrum beta-lactamase-producing Escherichia coli. AB - This study compared the potential synergy of levofloxacin and ciprofloxacin in combination with cefepime, ceftazidime, imipenem, piperacillin/tazobactam or amikacin, against extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli by using checkerboard and time kill studies. Moreover, selection of resistance was determined by frequency of mutations and by calculating the increase in minimum inhibitory concentrations (MICs) after five serial subcultures on antibiotic-containing plates. Synergy occurred more often with levofloxacin combined with imipenem (7/10 strains) and with levofloxacin or ciprofloxacin with amikacin (10/10) than for the other combinations. Time kill studies showed synergy for levofloxacin combined with amikacin, ceftazidime, imipenem or piperacillin/tazobactam, and for ciprofloxacin combined with amikacin, cefepime or imipenem. Antibiotic combinations selected for resistance less frequently than antibiotics alone. Mutation frequency was <10(-12) for all combinations. In conclusion, the combination of a fluoroquinolone with a beta lactam or amikacin may provide improved antimicrobial activity and help limit the occurrence of resistance in ESBL-producing E. coli strains. PMID- 15828444 TI - Antimicrobial resistance of nosocomial strains of Staphylococcus aureus in Russia: results of a prospective study. AB - A total of 879 Staphylococcus aureus clinical isolates from 17 medical institutions in different regions of Russia were tested. Susceptibility to 18 antimicrobials was determined by agar dilution in accordance with the NCCLS recommendations. The most potent antimicrobials were glycopeptides, linezolid, and fusidic acid, to which no resistance was found. Other antimicrobials with low frequency of non-susceptibility were mupirocin (0.3%), trimethoprim/sulfamethoxazole (0.8%), quinupristin/dalfopristin (1.8%) and rifampicin (7.0%). Fluoroquinolones displayed moderate activity (5.8% of non susceptible strains to moxifloxacin, 9.1% to levofloxacin, 13.1% to ciprofloxacin). High rates of non-susceptibility were found to clindamycin (27.1%), gentamicin (30.7%), tetracycline (37.1%), erythromycin (39.6%) and chloramphenicol (43.1%). The prevalence of oxacillin-resistant S. aureus (ORSA) was 33.5% and varied from 0% to 89.5% in different hospitals. ORSA were isolated most frequently in the burn units (77.5%), intensive care units (54.8%), trauma and orthopedics units (42.1%). This is the first multicenter study published of antimicrobial resistance of S. aureus in Russia which meets international standards. PMID- 15828445 TI - Gram-negative bacterial resistance to cephalosporins in community-acquired infections in Turkey. AB - Cephalosporins are widely used and trustworthy antibiotics in daily medical practice. Although antibacterial resistance has been reported in hospital wards, there are less data for community-acquired infections. In this study we investigated the cephalosporin susceptibility profiles of community-acquired Gram negative bacteria isolates in Sivas Kizilay Medical Center (Turkey) between March 2002 and March 2003. In our study, 949 Escherichia coli, 165 Proteus spp., 97 Enterobacter spp., 24 Klebsiella spp and 84 Pseudomonas aeruginosa strains were evaluated. Cefepime seemed to be the most effective antibiotic against our community-acquired Gram-negative isolates. Resistance to this drug was 19.3% for P. aeruginosa and around 0-10.6% for enteric bacteria. Enteric pathogen resistance ranged between 44.3-100% for cefazolin, 25-51.9% for cefuroxime, 4.8 25.3% for ceftriaxone, 5.4-14.5% for ceftazidime. Resistance in enteric pathogens to gentamicin ranged between 5.8-15.4%, to amikacin between 3.8-6.25%, to ciprofloxacin between 6.7-20%. 8.1% of P. aeruginosa were resistant to ciprofloxacin. With these profiles the aminoglycosides and ciprofloxacin resemble highly effective cephalosporins like cefepime. On the contrary, first- and second generation cephalosporins, trimethoprim-sulfamethoxazole, ampicillin and ampicillin-sulbactam are no longer used in probable Gram-negative bacterial infections in our region. Since treatment based on cephalosporins was less efficacious than expected in community-acquired infections, urgent measures are needed to limit antibacterial resistance outside of hospitals. PMID- 15828446 TI - Extended Spectrum Beta-Lactamase-producing Klebsiella pneumoniae outbreaks during a third generation cephalosporin restriction policy. AB - In spite of the adoption of third generation cephalosporin restriction policies, two independent outbreaks by Extended Spectrum Beta-Lactamase (ESBL)-producing Klebsiella pneumoniae occurred in two different wards (Neonatal Intensive Care Unit, NICU and Neurosurgery) of a teaching hospital in Rome, Italy. In the former 19 infected neonates were reported, whereas in the latter there were 10 infected patients. In both wards no differences were observed in the mortality rates in periods of outbreak and those with no outbreak. Molecular typing on a total of 19 isolated strains was carried out and restriction patterns were compared. The PFGE showed that nine isolates responsible for infection in the NICU were all included in three closely related clusters. In Neurosurgery nine strains out of ten were strictly related and part of an outbreak occurring between August-December 2003, while one isolate was temporarily (February 2003) and genetically (seven band differences) unrelated to the outbreak strains. When ESBL producing K. pneumoniae clusters from the two wards (NICU vs Neurosurgery) were compared, they appeared to be completely different both for their genotype pattern and plasmid type or presence, thus demonstrating cross transmission by two different genotypes. PMID- 15828447 TI - Strategic alliance between the infectious diseases specialist and intensive care unit physician for change in antibiotic use. AB - There is a general consensus that antimicrobial use in intensive care units (ICU) is greater than that in general wards. By implementing a strategy of systematic infectious disease consultations in agreement with the ICU chief, we have modified the antibiotic prescription habits of the ICU physician. A reduction was observed in the use of selected antibiotics (third-generation cephalosporins, vancomycin, carbapenems and piperacillin-tazobactam), with a significant reduction in the length of hospital stay for ICU patients and lower antibiotic costs without negative impact on patient mortality. Leadership by the infectious diseases consultant in combination with commitment by ICU physicians is a simple and effective method to change antibiotic prescription habits in the ICU. PMID- 15828448 TI - Antibiotic-associated diarrhea in a Turkish outpatient population: investigation of 288 cases. AB - Oral antibiotics are often prescribed, especially for respiratory tract infections in the community. The widespread use of broad-spectrum antibiotics causes an increased incidence of antibiotic-associated diarrhea (AAD). Although AAD has been studied in hospitalized patients, there is little available information concerning the characteristics of AAD in outpatient populations. The aim of this study was to investigate the clinical and laboratory findings of adult patients with community-acquired AAD. Between June 1998 and December 2003, the clinical reports of 288 patients were retrospectively reviewed. We observed that the duration between the start of antibiotic treatment and onset of symptoms was 7 days in most of the patients (86%), and the mean time was 9+/-1.0 days. The diarrhea was self-limited in all cases and mean duration of symptoms was 3 (+/ 1.0) days (1-7 days). The most common symptoms were abdominal discomfort and tenesmus (61.1%), while elevated WBC counts and fever were detected rarely. We were able to perform microbiologic investigations in only 88 patients because of the financial problems. Of the 88 stool specimens tested, none of them were positive for pathogenic bacterial growth or toxin A production. PMID- 15828449 TI - Penetration of amphotericin B in human lung tissue after single liposomal amphotericin B (AmBisome) infusion. AB - The distribution of amphotericin B in lung tissue was studied in 18 patients with primary or secondary lung cancer who underwent thoracotomy and pulmonary resection. At different times before surgery the patients were treated with liposomal amphotericin B 1.5 mg/kg by i.v. infusion over 1h. Blood and lung tissue samples were collected during surgery (one subject for each collecting time) and assayed for amphotericin B levels by HPLC. Due to surgical requirements, it was possible to obtain data from the 10th to the 25th h after the end of infusion. Plasma amphotericin B concentrations progressively decreased from 3.4 microg/ml at the 10th h to 1 microg/ml at the 25th h after the end of intravenous infusion. In lung tissue samples the lowest amphotericin B concentration (about 1 microg/g) was observed at the 10th h, then a progressive increase was observed with the highest value (2.5 microg/g) determined at the 25th h. PMID- 15828450 TI - Inhibition of P-glycoprotein function and expression by kaempferol and quercetin. AB - The 170 kDa plasma membrane P-glycoprotein (Pgp) causes the efflux of chemotherapeutic drugs from cells and is believed to be an important mechanism in multidrug resistance (MDR) in human cancer. This study demonstrates that some putative flavonoids, i.e., flavonols (quercetin and kaempferol) and isoflavones (genistein and daidzein) markedly increase the sensitivity of the multidrug resistant human cervical carcinoma KB-V1 cells (high Pgp expression) to vinblastine and paclitaxel dose-dependently, and also decrease the relative resistance of these anti-cancer-drugs in KB-V1 cells. None of the flavonoids had a significant effect on vinblastine and paclitaxel cytotoxicity in wildtype drug sensitive KB-3-1 cells (lacking Pgp). These flavonoids also caused an increase in intracellular accumulation, and reduced the efflux of Rh123 and 3[H]vinblastine in KB-V1 cells, but not in KB-3-1 cells. The flavonols increased the inhibitory effectiveness of Pgp activity in MDR KB-V1 cells more than isoflavones. Only treatment with flavonols up to 48 h was able to significantly decrease the Pgp expression in a dose-dependent manner in KB-V1 cells. These findings provide evidence that flavonols reduced Pgp expression and function resulting in the inhibition of Pgp activity, but isoflavones modulated intracellular drug levels by inhibiting Pgp function with no effect on Pgp expression. Among the flavonoids tested, flavonols, particularly kaempferol, exhibit the most potent MDR reversing property in KB-V1 cells. PMID- 15828451 TI - Potential predictive factors for response to weekly paclitaxel treatment in patients with metastatic breast cancer. AB - The authors compare results obtained from weekly paclitaxel treatment in advanced breast cancer patients with biological and clinical prognostic factors. Expression of c-erbB-2, Ki-67, p53 and hormone receptors (HR) was examined by immunohistochemistry in samples of breast tissue from 30 patients. Univariate analysis showed that Ki-67 positivity and low performance status (PS) were associated with poor outcome (P <0.05). We observed that expression of p53 and c erbB-2 did not have any negative effect on response to chemotherapy and survival. HR-negative patients had better response and slightly statistically significant overall survival (OS) rates compared to HR-positive patients (P >0.05). In a multivariate analysis low PS was the only significant predictor of shorter survival (P <0.05). In conclusion, while the expression of p53 and c-erbB-2 did not have any effect on treatment results, negative Ki-67 expression and negative HR status were associated with better OS in this patient population. PS was the only significant predictor for OS. PMID- 15828452 TI - Carboplatin hypersensitivity reactions: a single institution experience. AB - Carboplatin-related hypersensitivity reactions, frequently encountered in the heavily pretreated subpopulation of patients with gynecologic malignancies, can be severe and even potentially lethal-precluding these patients from an effective salvage treatment. We describe our experience in the management of such reactions and the application of a pretreatment protocol with corticosteroids, antihistamines and a slow infusion rate in order to safely re-administer carboplatin to the above patients. From 1998 to 2004, twenty patients developed an allergic reaction to carboplatin. Sixteen of them (80%) suffered from ovarian cancer. Upon resolution of the acute reaction, thirteen patients were pretreated according to our protocol and were re-exposed to carboplatin. Fifteen patients experienced the reaction during second-line carboplatin-based treatment and 5 patients after 3 or more regimens. Fifteen of the reactions (75%) were severe. Thirteen patients were re-treated with carboplatin after the application of our protocol, all of them successfully, even though 10 patients (77%) experienced minor symptoms during subsequent courses. On the contrary, only one of the 6 patients who were re-treated without the application of the protocol was able to receive further platinum-based treatment. In conclusion, pretreatment with corticosteroids, antihistamines and a slower infusion rate may make re-treatment possible in patients having experienced hypersensitivity to carboplatin. PMID- 15828453 TI - Treatment of a Gilbert's syndrome patient with irinotecan, leucovorin and 5 fluorouracil. AB - The patient was an elderly male who had radical surgery for sigmoid cancer in 2001. Owing to metastasis of his cancer to the left supraclavicular lymph nodes in 2002, the patient was admitted to our hospital for systemic chemotherapy. We started treatment with irinotecan, leucovorin, 5-fluorouracil (IFL). After administering 100 mg/m2 of irinotecan, 250 mg/m2 leucovorin and 600 mg/m2 5 fluorouracil to the patient on day 1, grade 3 leukopenia developed rapidly and grade 4 thrombocytopenia was observed on day 5. We excluded irinotecan from the medication and continued the administration of 5-fluorouracil and leucovorin, but his tumors had not been reduced sufficiently. Based on some examination results, we assumed that the patient had Gilbert's syndrome and that the severe side effects that occurred were due to prolongation of SN38 metabolism. We again administered irinotecan but at reduced dose (25 mg/m2). Four courses of this modified IFL were administered safely and the response was favorable. PMID- 15828454 TI - Empiric antifungal strategies for acute leukemia patients. PMID- 15828455 TI - Antimicrobial sensitivity of Neisseria gonorrhoeae: a hospital-based study from Kuwait. PMID- 15828456 TI - [Late stent-thrombosis-related myocardial infarction]. AB - The objective of the study was to assess the frequency of late stent-thrombosis related acute myocardial infarction in patients with LAD stents. A series of 930 PCI procedures involving LAD stenting were performed in 920 patients (mean age: 63 years, 667 men) from January 1999 to December 2001. Among those 14 patients had late (> 30 days), thrombosis-related acute myocardial infarction after a mean delay of 135 +/- 131 days. Three-year thrombosis-free survival was 97.6%. By univariate analysis, risk of late thrombosis was associated with LAD-diagonal bifurcation stenting, placement of a stent on the ostium of a diagonal branch and number of stents implanted on the LAD. Patients with these risk factors might be appropriate candidates for long-term combined antiplatelet therapy. PMID- 15828457 TI - [Prehospital management of very elderly patients with ST segment elevation in Paris by mobile intensive care units (Samu)]. AB - More and more elderly people are hospitalised with myocardial infarction. Little is known on their pre-hospital management. In 2001 and 2002, 105 patients aged 80 years or more with suspected ST elevation infarction were managed by the mobile intensive care unit system of the SAMU de Paris-Necker. Diagnosis of infarction was confirmed in 92 (88%). Over 60% of the patients were women. Median time delay from symptom onset to call to the emergency service was 127 minutes, longer in nonagenarians (175 vs 101 minutes). Prehospital use of aspirin was 81% and 39% received an intravenous bolus of heparin. A reperfusion strategy was decided in only 30% (primary PCI: 23/26). One-month mortality was 21% and was related to older age, time when the call to the Samu was made, and absence of current smoking. Overall, the prehospital management of very elderly patients with suspected ST elevation infarction appears far from optimal. PMID- 15828458 TI - [Impact of diabetes on mortality after myocardial infarction]. AB - Myocardial infarction is a common cause of mortality in people with diabetes. The aim of this study was to determine early and mid-term mortality in diabetic patients with myocardial infarction and to determine if hyperglycemia was predictor of mortality. We conducted a retrospective study of 100 diabetic patients compared with 100 non diabetic patients who were hospitalised in our institution between 1999 and 2003 for myocardial infarction. RESULTS: Hospital and one year mortality were highest among diabetic patients compared with non diabetic patients. Multivariate analysis showed that admission plasma glucose was a consistent predictor factor of in hospital mortality RR 1.2 (IC 1.02-1.47). Admission plasma glucose was significantly higher in nonsurvivors diabetic patients than in survivors (22.7 vs 16 mmol/l P = 0.04). The predictor factors of one year mortality was age, female sex and no beta blocker at discharge RR5.3 (1.9-14.3). CONCLUSION: Diabetic patients with myocardial infarction have poor prognosis and hyperglycemia was associated with in hospital mortality. PMID- 15828459 TI - [Retrospective survey of the management of patients treated for acute myocardial infarction in Provence-Alpes-Cote d'Azur]. AB - OBJECTIVE: The aim of the study is to assess the characteristics, outcomes and factors associated with delay of reperfusion therapy in patients with myocardial infarction in the Provence Alpes Cote d'Azur (PACA). METHODS: This retrospective study included all patients hospitalized with myocardial infarction in the PACA between January and June 2000. Myocardial infarction patients were identified using the Programme de Medicalisation des Systemes d'Information. Univariate and multivariate analysis were performed. RESULTS: A total of 2049 patients were admitted in 74 hospitals. Mean patient age was 68.9. Treatment consisted of coronary angioplasty alone in 53.3% of cases, thrombolysis alone in 4.5% and angioplasty after thrombolysis in 7.7%. The complication rate was 27.4% and mortality was 9.5%. The median time from first awareness of symptoms to the first action taken by the patient (1) was 1 hour 30 minutes. The time from the first action taken by the patient to revascularisation (2) was 3 hours 30 minutes. The time between the first awareness and revascularisation (3) was 8 hours 45 minutes. Multivariate analysis showed that the following factors were associated with increased delay: (1). no prior myocardial infarction, occurrence of symptoms at home. (2). age > 65 years, absence of chest pain, consultation with a physician before hospitalization, non-medical transport, transfer from one hospital to another, treatment by angioplasty alone. (3). age > 65 years, absence of chest pain, consultation with a physician before hospitalization, non-medical transport, treatment by angioplasty alone. CONCLUSIONS: Alerting emergency services more quickly would shorten treatment delay not only by ensuring quicker transport to a properly equipped hospital but also by allowing prompt pre hospital thrombolytic therapy. PMID- 15828460 TI - [Management of acute coronary syndromes in a new French coronary intensive care unit. The first four years of activity in the GRACE registry (Global Registry of Acute Coronary Events)]. AB - The GRACE registry provides the opportunity to analyse management of acute coronary syndromes in the real word and the impact of hospital characteristics. In this setting, we compare the activity of a new coronary intensive care unit with regional data. METHODS: From January 2000 to December 2003, 376 eligible patients were involved (22% of regional inclusion). GRACE standard diagnosis were the following, for our centrer (for the cluster): ST elevation myocardial infarction 28% (37%), non-ST elevation myocardial infarction 32% (31%), unstable angina 33% (24%). Demographic characteristics were similar with a median age of 64 (vs 66) and a large majority of male (74 vs 81%). Medical history and cardiovascular risk factors were comparable. Predictors of hospital mortality were observed at the same rate: cardiogenic shock (3 vs 3%), congestive heart failure > Killip 2 (4 vs 4%), left ventricular ejection fraction (LVEF) lower than 40% (17 vs 16%), recurrent ischemic symptoms (8 vs 8%). Coronary artery bypass grafts were required in 5% (vs 2%). RESULTS: Drugs prescription rates were similar: aspirin at admission (95%) and at discharge (95%), betablocker at admission (70%) and at discharge (85%), statin at admission (< 30% in 2000, > 60% in 2003) and at discharge (< 60% in 2000 and > 80% in 2003), ticlopidin clopidogrel at admission (< 20% in 2000 and > 40% in 2003), ACE inhibitor for LVEF < 40%, intravenous GPIIblIIa, and low molecular weight heparin (90%). Cardiac catheterisation (90%) and percutaneous coronary interventions (80%) were performed at the same rates in our center and in the cluster. Hospital death was similar (2 vs 4%). Discharge status was home for a large majority of patients (63 vs 76%). The median length of stay was five days and shorter than three days for patients with unstable angina. CONCLUSION: Based on GRACE registry data, the present evaluation revealed that our new center offered evidence-based medical and interventional therapy in patients with acute coronary syndromes at the same level than experienced institutions with similar results for hospital death and length of stay. PMID- 15828461 TI - [Mortality prognostic factors of cardiogenic shock complicating an acute myocardial infarction and treated by percutaneous coronary intervention]. AB - OBJECTIVE: To determine the in-hospital prognosis and late outcome of cardiogenic shock complicating acute myocardial infarction treated by early (< 24 hours) percutaneous coronary intervention (PCI). METHODS: Retrospective monocentric study of a consecutive cohort of patients undergoing early PCI (< 24 heures) for cardiogenic shock complicating acute myocardial infarction from 1994 to 2004. RESULTS: The cohort included 175 patients (mean age = 65 +/- 14 years, 68% male). A successful PCI was obtained in 69% of patients. The in-hospital mortality was 43%. Independent risk factors associated with an increased mortality were: absence of TIMI three flow (P < 0.0001), absence of smoking (P < 0.009) and the need for mechanical ventilation (P < 0.002). Nor stent use or anti GP IIb/IIa infusions were predictors of a better outcome. At hospital discharge, mean left ventricular ejection fraction (LVEF) was 38 +/- 12%. Kaplan-Meier estimate of survival was 63% for in-hospital survivors (maximum follow-up = 9 years). Independent predictors of an impaired long-term outcome were: a LVEF < 0.3 (P < 0.028) and 3-vessel disease on coronary angiography (P < 0.004). CONCLUSION: In hospital mortality of patients suffering cardiogenic shock complicating acute myocardial infarction and treated by PCI remains high despite PCI improvement. The long-term survival appears, however, to be better than that of patients with coronary artery disease and low LVEF. PMID- 15828462 TI - [Long-term follow-up after primary angioplasty: is stenting beneficial?]. AB - BACKGROUND: Primary stenting leads to a better short-term outcome than balloon angioplasty for acute myocardial infarction in randomised trials. However few data are available about the long-term outcome of primary stenting in acute myocardial infarction (AMI). OBJECTIVES: The aim of this study was to compare the three-year outcome after primary stenting versus balloon angioplasty in patients with acute myocardial infarction. METHODS: We conducted a retrospective study including 157 patients with AMI in a single center. Patients underwent balloon angioplasty (N = 48) or primary stenting (N = 109) within six hours after the onset of chest pain. We looked at the outcome during three years focusing on global mortality, major adverse cardiac events (MACE), reinterventions and target vessel revascularization (TVR). RESULTS: The two groups are similar for their baseline characteristics. No difference was noted for in-patient mortality in the balloon angioplasty group and the primary stenting group (2.1 vs 2.8%; P = ns). The three-year mortality was not significantly different in the two groups. Regarding MACE (27.8 vs 31.7; P = 0.95), reinterventions (20.4 vs 24.7%; P = 0.98) and TVR (18.6 vs 17.8%; P = 0.69), both groups were statistically not different. CONCLUSION: In the long-term patients treated with stent placement have similar rates of MACE, reinterventions or TVR than patients undergoing balloon angioplasty. If few studies noted a benefit in short-term outcomes, primary stenting doesn't improve the prognosis of acute myocardial infarction on long-term follow-up, which is dependent on atherosclerosis. PMID- 15828463 TI - [The vascular manifestations of Behcet's disease: a case report]. AB - Behcet's disease is an inflammatory vasculitis which affects the arteries and veins. The vascular pathologies are the rare complications of this disease. We present here a patient with Behcet's disease who has been hospitalized several times because of plurifocal vascular manifestations. PMID- 15828464 TI - [Cardiac and vascular effects of cannabinoids: toward a therapeutic use?]. AB - Interest in cannabinoid pharmacology developed rapidly since the discovery of cannabinoids receptors and endocannabinoids. Modulation of this system is becoming a hot topic in cardiovascular pharmacology mainly at the light of recent findings. Among them, cardiac effects of cannabinoids were described with respect to their probable participation to the well-studied preconditioning phenomenon. Beneficial effects of post-infarction cannabinoids administration against ischemia-reperfusion injury were also reported. Concerning their vascular effects the situation is more complex some studies reporting pressor effects while others depressor ones. It was also proposed that the endothelium-derived hyperpolarizing factor released by various vasodilators may be an endocannabinoid an hypothesis still discussed. Finally, pathological situations concerning the cardiovascular system and including brain ischemia, hemorrhagic and endotoxic shocks were reported to be linked with endocannabinoids. However, the clinical use of cannabinoid receptors agonists or antagonists will depend on the development of non psychoactive compounds. PMID- 15828465 TI - [Acute myocarditis: from chest pain to cardiogenic shock]. AB - We report the observations of three patients with myocarditis. The first one with chest pain, the second one with a pseudo-infarct presentation and the third one with a cardiogenic shock. We discuss the different anatomo-clinical presentations of myocarditis, the diagnosis, the indications of endomyocardial biopsies and the prognosis of this pathology. PMID- 15828466 TI - [Study on in vivo drug delivery and repairing large segmental infected bony defect with massive reconstituted bovine xenograft aided by calcium phosphate cement drug core]. AB - OBJECTIVE: To find out an effective technique to repair large segmental infected bony defect. METHODS: Calcium phosphate cement (CPC) incorporated with bone morphogenetic protein and gentamycin was embedded in the massive reconstituted bovine xenograft (MRBX), then CPC-MRBX was obtained after CPC's solidification. In vivo test was applied to test the drug delivery capability of CPC-MRBX, in which it was implanted in the dorsal muscle pouch of 18 rabbits. The drug concentration of animal blood and surrounding soft tissue of the CPC-MRBX in the muscle pouch was measured 1, 2, 5, 10, 15, 20, 25, 30 and 35 d after operation, 2 rabbits each time. Large segmental infected femur defect in the rabbit model was created to test the repairing capability of CPC-MRBX. External fixation was done 1.5-2.0 cm above the knee, the most adjacent nail to fracture site was 0.5-0.8 cm away, and proper pressure was applied to the graft. In experimental group (n = 25), the bony defect was replaced by CPC-MRBX, while in the control group(n = 15) dissected bone block was re-implanted in original position. The animal was subjected to radiographic, histological examination at 4, 8, 16 and 24 weeks. The general condition was observed after the operation. RESULTS: CPC-MRBX was easily made under normal temperature and pressure. In vivo drug delivery test showed that the drug concentration of the tissue remained above the minimal inhibitory concentration of staphylococcus 30 d after operation and no significant increase of blood drug concentration was observed. In experimental group, no adverse influence was observed. Four weeks after operation, the animal could bear load, bony callus around the graft was observed by X-ray, and abundant chondral tissues that grew into CPC-MRBX were observed by histological method. Eight weeks after operation, progressively increasing bony callus around the graft was observed, external fixation could be removed, normal function was restored, and CPC was degenerated dramatically while new bone tissues were growing. Sixteen weeks after the operation, more new bone tissues grew and CPC was degenerated further while marrow tissues were taking shape. Twenty-four weeks after the operation, femur healed completely and CPC was degenerated completely. In the control group, the autograft remained unhealed on X-ray at 4 weeks, and osteomyelitis manifestation such as inflammatory cells infiltration and osteolysis was detected at 4 weeks. All the animals in the control group died before the 8th week, 4 of which showed positive hemoculture. CONCLUSION: CPC-MRBX is readily available and can be applied to repairing large segmental infected bony defect. PMID- 15828467 TI - [Experimental study on the treatment of femur head necrosis with tricalcium phosphate and platelet-rich plasma]. AB - OBJECTIVE: To study the mechanism of compound of calcium phosphate (TCP) and platelet-rich plasma(PRP) in the treatment of femoral head necrosis. METHODS: The left femoral heads of 48 New Zealand white rabbits were frozen by liquid nitrogen as to make the model of femoral head necrosis. Twenty-four rabbits were randomly chosen as the experimental group and their femoral heads were filled with TCP/PRP. The other 24 rabbits were used as the control group and their femoral heads were filled only with TCP. They were sacrificed at 2, 4, 8, 12 weeks after operation. The specimens were examined with X-ray and histological study. RESULTS: At 2 weeks after operation, there was no significant difference in femoral head density between the two groups. Four weeks after operation, femoral head density decreased in both groups, while it decreased more in the control group. At 8, 12 weeks after operation, the density of the femoral heads in both groups increased, and it was higher in the experimental group. Histology examination showed that there was no difference between the two groups 2 weeks after operation. The head became flat at 4 weeks. Control group had more defects. At 4, 8, 12 weeks, more repairs were observed in the experimental group than that in the control group. The amount and maturity of osteogenesis in experimental group were much more greater than those in control group. Bone histomorphometry showed that the volume of the trabecular was larger in the experimental group (36.65% +/- 7.22%, 38.29% +/- 4.28%, 39.24% +/- 3.42%) than that of control group (P < 0. 05). CONCLUSION: TCP/PRP does not only provide osteoblasts scaffold, but also promotes bone formation and the head repair. TCP/PRP is a good biomaterial for the treatment of femur head necrosis. PMID- 15828468 TI - [Experimental study of the effect of new bone formation on new type artificial bone composed of bioactive ceramics]. AB - OBJECTIVE: To investigate the osteogenic potential of four kinds of new bioactive ceramics combined with bovine bone morphogenetic proteins (BMP) and to explore the feasibility of using compounds as bone substitute material. METHODS: Ninety six rats were divided into 4 groups (24 in each group). BMP was combined with hydroxyapatite (HA), tricalcium phosphate (TCP), fluoridated-HA (FHA), and collagen-HA(CHA) respectively. The left thighs of the rats implanted with HA/BMP, TCP/BMP, FHA/BMP, and CHA/BMP were used as experimental groups. The right thighs of the rats implanted with HA, TCP, CHA, and decalcified dentin matrix (DDM) were used as control groups. The rats were sacrificed 1, 3, 5 and 7 weeks after implantation and bone induction was estimated by alkaline phosphatase (ALP), phosphorus (P), and total protein (TP) measurement. The histological observation and electronic microscope scanning of the implants were also made. RESULTS: The cartilage growth in the 4 experimental groups and the control group implanted with DDM was observed 1 week after operation and fibrous connective tissues were observed in the other 3 control groups. 3 weeks after implantation, lamellar bone with bone marrow and positive reaction in ALP stain were observed in the 4 experimental groups. No bone formation or positive reaction in ALP stain were observed in the control groups. The amount of ALP activity, P value, and new bone formation in the experimental groups were higher than those in the control group(P < 0.05). The amount of ALP activity, P value, and new bone formation in TCP/BMP group were higher than those in HA/BMP, CHA/BMP and FHA/BMP groups (P < 0.05). There was no significant difference in TP between the BMP treatment group and the control groups. From 5th to 7th week, new bone formation, histochemistry evaluation, and the level of ALP, P, TP value were as high as those in the 3rd week. CONCLUSION: New composite artificial bone of TCP/BMP, HA/BMP, CHA/BMP, and FHA/BMP all prove to be effective, but TCP/BMP is the most effective so that it is the most suitable biomaterial replacement of tissue. PMID- 15828469 TI - [Platelet-rich plasma made by a modified method promotes proliferation of rat osteoblast and human osteoblast in vitro]. AB - OBJECTIVE: To study the effect of serum rich in growth factors (SRGF) derived from platelet-rich plasma (PRP) on the biological function of human and rat osteoblast. METHODS: PRP and platelet-poor plasma (PPP) obtained from healthy human and SD rat were activated by thrombin to get SRGF and serum poor in growth factors (SPGF). The level of TGF-beta1 and PDGF-AB in human-SRGF and SPGF were assayed by enzyme-linked immunoassay (ELISA). Rat and human osteoblast were cultured and identified. Rat osteoblasts were treated with 5% rat-SRGF, 5% rat SPGF and serum-free F12 medium, respectively. And human osteoblast were treated with 5% human-SRGF, 5% human-SPGF and serum-free DMEM. Cellular mitogenic activity was evaluated by thiazolyl blue (MTT) colorimetric assay at 24, 48, 72 and 96 hours. RESULTS: The level of TGF-beta1 in human-SRGF was 307.67 +/- 35.57 ng/ml, and that of PDGF-AB was 52.76 +/- 7.89 ng/ml. The proliferation of rat and human osteoblast were promoted after treated with rat-SRGF and human-SRGF, respectively. In rat osteoblast groups, there were significant differences in absorbency between rat-SPGF group and rat-SRGF group at 48 and 96 hours (P < 0.05). In human osteoblast groups, the differences between human-SPGF group and human-SRGF group were significant at 48, 72 and 96 hours (P < 0.05). The proliferation of these two kinds of osteoblasts almost stopped in serum-free medium, and the differences in absorbency, compared with other groups, were significant (P < 0.05). CONCLUSION: High quality of PRP can be achieved by the improved method and SRGF is capable of up-regulating the proliferation of rat osteoblast and human osteoblast. PMID- 15828470 TI - [Study on the effect of composite of basic fibroblast growth factor and partially deproteinized bone on the repair of femoral head defects]. AB - OBJECTIVE: To evaluate the effect of composite (bFGF/PDPB) of basic fibroblast growth factor (bFGF) and partially deproteinized bone (PDPB) on the repair of femoral head defect. METHODS: Forty-eight femoral heads with defect derived from 24 New Zealand rabbits were divided into 3 groups at random, which were implanted with bFGF/PDPB (group A), PDPB (group B) and nothing (group C) respectively. The rabbits were sacrificed at 2, 4, and 8 weeks after operation, and then the femoral heads were obtained. The specimens injected with Chinese ink were created. Then X-ray examination, histopathological and morphological examination of blood vessel, and image analysis were made. RESULTS: The bone defects healed completely 8 weeks after operation in group A. The implants in the repaired tissue were not substituted completely in group B. The bone defects did not heal completely in group C. Two weeks after operation, affluent newly formed vessels were seen in repaired areas in group A. No significant difference between group A and group B was observed 8 weeks after operation. In group C, newly formed vessels were scarce 2, 4, and 8 weeks after operation. There were 3 sides rated excellent, 2 good and 1 fair in group A; 1 excellent, 2 good, 2 fair and 1 poor in group B; and 1 fair and 5 poor in group C according to the X-ray evaluation 8 weeks after operation. Eight weeks after operation, the volume fraction of bone trabecula in repaired tissue was higher in group A than that in group B (P < 0.05), and the fraction in group C was the lowest among the 3 groups (P < 0.05). CONCLUSION: The composite of bFGF and PDPB can effectively promote the repair of femoral head defect of rabbit. PMID- 15828471 TI - [Combined use of autologous micro-morselized bone with bone morphogenetic protein and type I collagen graft in repairing rabbit bone defects]. AB - OBJECTIVE: To study the effect of combined use of autologous micro-morselized bone with bone morphogenetic protein(BMP) and type I collagen graft on the treatment of segmental bone defects. METHODS: The bulk bone of rabbit iliac crest was ground into micro-morselized bone, which was combined with BMP and type I collagen. The model of 1.5 cm bone defect was established in the middle shaft of the radius. Fifty-six rabbits were assigned to four repairing methods: autologous micro-morselized bone graft with BMP and type I collagen, autologous micro morselized bone graft with type I collagen, autologous micro-morselized bone graft alone, and control group. The defect-repairing capability of each group was assessed by radiographic, histological, bone densitometry and biomechanical studies. RESULTS: X-ray manifested that at the end of 8 weeks after operation, the bone defect treated with autologous micro-morselized bone graft with BMP and type I collagen was repaired completely, and at the end of 12 weeks after operation the bone defect treated with autologous micro-morselized bone and type I collagen was cured completely, but the bone defect treated with autologous micro-morselized alone was completely repaired. No healing was found in the control group. In the bone densitometry detection, the material with BMP exhibited the strongest defect-repairing capability in terms of amount increased and quality of the new bone at the end of 8 weeks and 12 weeks. The group with BMP has the best mechanical strength of all groups at the end of 12 weeks. CONCLUSION: Autologous micro-morselized bone graft with BMP/type I collagen and autologous micro-morselized bone graft with type I collagen prove to be effective in repairing segmental bone defects. The autologous micro-morselized bone combined BMP and type I collagen is an excellent bone repairing material considering the satisfactory osteogenesis, osteo-conduction, and osteo-induction seen in this method. PMID- 15828472 TI - [Femoral varus osteotomy combined with interlocking nailing for treatment of genu valgum]. AB - OBJECTIVE: To assess the effect of medial distal femoral osteotomy combined with interlocking nailing on the treatment of knee osteoarthritis with valgus deformity. METHODS: From May 1996 to August 2000, 16 patients with knee osteoarthritis accompanied by valgus deformity were treated by medial wedged distal femoral osteotomy combined with interlocking nailing. Full-length radiographs were taken before operation and 8 weeks and 2 years after operation. The parameters, including the femorotibial angle, the tibial angle, the femoral angle, the femoral condyle-tibial plateau angle, and the lateral joint space, were measured by these radiographs. The function of knee was evaluated by the 100 point rating scale standard of knee. RESULTS: The mean postoperative score was significantly improved from 50.4 +/- 15.9 points to 78.5 +/- 12.9 points 2 years after the surgery. The lateral joint space was increased from 2.1 +/- 1.8 mm to 4.7 +/- 1.7 mm and the femoral condyle-tibial angle decreased from 5.6 +/- 2.9 degrees to 1.6 +/- 3.4 degrees. There were complications in 2 cases: 1 case of delayed union and 1 case of superficial wound infection. CONCLUSION: Medial distal femoral osteotomy combined with interlocking nailing proves to be an effective approach to treat knee osteoarthritis with valgus deformity. PMID- 15828473 TI - [The treatment of middle and lower thirds fractures of humerus by anterior plate fixation]. AB - OBJECTIVE: To study the effect of anterior plate fixation on the treatment of middle and lower thirds fractures of humerus and the possibility of operating without injuring the radial nerve. METHODS: Forty-nine patients with the middle and lower thirds fractures of the humerus were treated with anterior plate fixation from March 1998 to December 2002. Of the 49 patients, 27 were with new fractures, 12 with old fractures, and 10 with nonunion fractures. According to AO classification, of the 49 patients, there were 19 type A, 14 type B1, 9 type B2, 5 type B3, 2 type C1. Thirty-seven patients had closed fractures and 12 had open fractures. Of the 12 patients, 8 were type Gustilo I , 4 were type Gustilo I . Four out of the 49 patients were associated with radial nerve palsy. All patients were treated with anterior plate fixation through the anterior approach to the humerus. The radial nerves injured were explored. RESULTS: Of the 49 patients, 48 were followed up 6 to 48 months (28.7 months on average). All fractures were healed within 3 to 9 months(4.7 months on average). Fixed plates of 37 patients out of the 49 were removed. No iatrogenic radial nerve injury occurred in the 82 times of the operations among the 49 patients. CONCLUSION: The treatment of middle and lower thirds fractures of humerus by anterior plate fixation through the anterior approach to the humerus does not interfere with the fracture healing and can prevent the iatrogenic radial nerve injury. PMID- 15828474 TI - [The clinical effect of bipolar long-stem prosthetic replacement on the treatment of comminuted intertrochanteric fracture of hip in the elderly osteoporotic patients]. AB - OBJECTIVE: To evaluate the clinical effect of bipolar long-stem prosthetic replacement on the treatment of comminuted intertrochanteric fracture of hip in the elderly osteoporotic patients. METHODS: From March 2000 to August 2003, 18 patients who were more than 72 years old with comminuted intertrochanteric fracture were treated with bipolar long-stem prosthetic replacement. There were 8 males and 10 females (aged from 72 to 91). Fractured fragments of large trochanter and minor trochanter should be preserved during the operation so that the fragments can be fixed with steel wires and insertion of artificial femoral head. The surgical approach and operative technique, the average course of treatment in hospital, the average time to ambulate with full-weight bearing on the operated limb after the operation, complication, and mortality were observed. RESULTS: All patients were followed up 6 to 28 months (16.2 months on average). The average course of stay at hospital was 16 days. The average time to ambulate was 32 days. The incidence of the pain of the hip joint was one-eighteenth, and one patient due to another disease was unable to walk without using two crutches one month after the surgery. The mortality of the patients was one-eighteenth one year after the surgery. CONCLUSION: Bipolar long-stem prosthetic replacement for the treatment of comminuted intertrochanteric fractures in the elderly osteoporotic patients proves to be a suitable alternative. Patients have better prognosis, early full-weight bearing, rapid rehabilitation, and fewer complications. PMID- 15828475 TI - [Comparison of two different operations on comminuted intercondylar fracture]. AB - OBJECTIVE: To compare the effect of two different operations on treating severely comminuted intercondylar fracture. METHODS: From December 2001 to October 2003, 20 cases of severely comminuted intercondylar fracture were operated. Of the 20 cases, 7 (group 1) were treated with closed reduction and retrograde intramedullary nailing through arthroscope, 13 (group 2) were treated with open reduction and retrograde intramedullary nailing without arthroscope. Follow-ups were conducted after operation. RESULTS: All cases of fracture were recovered. Swelling in group 1 was alleviated more obviously than that in group 2. In group 1, all knees could flex to 120 degrees during 6th to 9th weeks after the operation. In group 2, only 4 could flex 110 degrees. CONCLUSION: Retrograde intramedullary nailing through arthroscope proves to be less invasive and more effective in treating heavily comminuted intercondylar fracture. PMID- 15828476 TI - [Clinical study of phalange fractures treated by absorbable intramedullary nail in replantation of severed finger]. AB - OBJECTIVE: To study the effect of internal fixation with absorbable intramedullary nail on the treatment of phalange fractures in replantation of severed finger. METHODS: From September 2001 to October 2003, 28 cases with industrial severed finger (21 males and 7 females, with the age of 18-35 years) were replanted within 1-6 hours. The severed locations were index fingers in 11 cases, middle fingers in 8 cases, ring fingers in 6 cases, little fingers in 3 cases. All cases of phalange fractures were fixed by absorbable intramedullary nails of poly-DL-lactic acid (PDLLA) that combined with chitosan. Out of the 28 cases, 15 cases were with proximal phalange, 11 cases were with middle phalange, 2 cases were with distal phalange. The bone marrow cavity of the phalanges were dilated, then the intramedullary nail was inserted with suitable diameter and length to fix the fracture. Postoperatively resin bandage was applied for 3-4 weeks. RESULTS: All the 28 patients survived the performance and postoperative follow-up ranged from 3 to 10 months (4 months in average). Rejection was observed in one case 3 weeks after operation, bone unions were obtained in other cases. According to the Chinese Medical Association's evaluation standard for replantation of amputated finger, 18 fingers resulted in excellence, 9 fingers in good function and 1 with unsatisfactory function. The excellent and good rate was 96.4%. CONCLUSION: Internal fixation with absorbable intramedullary nail of PDLLA combined with chitosan proves to be effective in the replantation of severed finger. PMID- 15828477 TI - [The application of reconstruction with autograft implantation in total hip replacement with regional acetabular deficiency]. AB - OBJECTIVE: To evaluate the effect of reconstruction with autograft implantation in total hip arthroplasty(THA) with regional acetabular deficiency. METHODS: From 1991 to 2000, 39 cases of THA with acetabular deficiency were conducted. Autogenous bone implantation was used to reconstruct the deficient acetabulum. Of the 39 patients, 25 were males and 14 were females. The age ranged from 34 to 62 (45.2 on average). There were 21 cases of developmental dysplasia resulted deficiency, 14 cases of fracture of femoral neck complicated with head necrosis (10 hips) and fracture of acetabulum(4 hips). The resected femoral heads or autologous ilium were made the wedge-shaped graft and implanted into the deficient acetabulum, which included 12 cases with cement THA and 27 with cementless THA. Of all the cases, 24 were followed up 2 to 10 years (6.7 years on average). Harris scores before operation were 18 to 50 (38.1 on average). RESULTS: The limbs were lengthened by 2.4 cm on average. No serious complications were observed in these patients. Compared with the scores before the operation, the average Harris scores after the operation were 92.1 (P < 0.01)and 86.3 (P < 0.05) in the one-year and the latest follow-up respectively. The rates for the good were 91.7% and 83.3% in the one-year and the latest follow-up respectively. CONCLUSION: The acetabular reconstruction with autograft in THA will bring better stability in those patients with acetabular deficiency. PMID- 15828478 TI - [Study on the effect of sodium hyaluronate intra-articular injection on the treatment of knee osteoarthritis]. AB - OBJECTIVE: To investigate the effect of sodium hyaluronate (SH) intra-articular injection on the treatment of knee osteoarthritis (OA), and to compare the contents of free radicals and inflammatory factors in joint fluids of pre- and pro-treatment as to explore the treatment mechanism of SH. METHODS: Ninety-two patients (111 knees) with mild(51), moderate (35) and serious (25) knee OA were treated with intra-articular injections of SH (20 mg once a week for 5 weeks). According to Lysholm scoring, clinical signs such as pain, swelling,and the ability to walk, squat, run, go upstairs and downstairs were assessed before and after the treatment, and the contents of nitric oxide (NO), superoxide dismutase(SOD), malonic dialdehyde (MDA) and IL-1beta, TNF-alpha in joint fluids from the OA joints before 1st, 2nd, and 5th injection and 3 months after each injection were observed. RESULTS: All cases were followed up for 3 months. The improvements in the signs and function of knees were excellent in 42 knees, good in 38 knees, fair in 21 knees and poor in 10 knees, with 72.1% excellent and good results. The lighter the illness was, the better the improvement was: the rate of the excellent and good was 92.1% in mild group, 68.6% in moderate group and 42.9% in serious group. The contents of oxygen free radicals and IL-1beta, TNF-alpha of the patients with mild and moderate OA decreased markedly after being treated with SH(P < 0.05), but these decreased lightly in serious OA group(P > 0.05). SH had mild effect on the contents of NO. Three months after treatment, only in mild OA group the contents of NO significantly decreased (P < 0.05), and no significant change in moderate and serious groups was observed (P > 0.05). CONCLUSION: SH intra-articular injection has a positive effect on the relief of clinical symptoms and on the improvement of articular function of knee OA. The therapeutical effect of SH on OA is achieved possibly by decreasing the contents of free radicals especially oxygen free radicals and inflammatory factors in joint fluids. PMID- 15828479 TI - [Monitoring retrograde adenoviral transgene expression in spinal cord and anterograde labeling of the peripheral nerves]. AB - OBJECTIVE: Targeted adenoviral gene delivery from peripheral nerves was used to integrally analyse the characterization and time course of LacZ gene (AdLacZ) retrograde transfer to spinal cord and transgene product anterograde labeling of peripheral nerve. METHODS: Recombinant replication-defective adenovirus containing AdLacZ was administrated to the cut proximal stumps of median and tibial nerves in Wistar rats. Then the transected nerve was repaired with 10-0 nylon sutures. At different time point post-infection the spinal cords of C5 to T1 attached with DRGs and brachial plexuses, or L2 to L6 attached with DRGs and lumbosacral plexuses were removed. The removed spinal cord and DRCs were cut into 50 microm serial coronal sections and processed for X-gal staining and immunohistochemical staining. The whole specimens of brachial or lumbosacral plexuses attaching with their peripheral nerves were processed for X-gal staining. The number of X-gal stained neurons was counted and the initial detected time of retrograde labeling, peak time and persisting period of gene expression in DRG sensory neurons, spinal cord motor neurons and peripheral nerves were studied. RESULTS: The gene transfer was specifically targeted to the particular segments of spinal cord and DRGs, and transgene expression was strictly unilaterally corresponding to the infected nerves. Within the same nerve models, the initial detected time of gene expression was earliest in DRG neurons, then in the motor neurons and latest in peripheral nerves. The persisting duration of beta-gal staining was shortest in motor neurons, then in sensory neurons and longest in peripheral nerves. The initial detected time of beta-gal staining in median nerve models was earlier in median nerve models compared with that in the tibial nerve models. Although the initial detected time and the beginning of peak duration of beta-gal staining were not same, the decreasing time of beta-gal staining in motor and sensory neurons of the two nerve models were started at about the same day 8 post-infection. The labeled neurons were more in tibial nerve models than that in median nerve models. Within the same models, the labeled sensory neurons of DRGs were more than labeled motor neurons of ventral horn. The beta-gal staining was tender in median nerves than that in tibial nerves. However the persisting time of beta-gal staining was longer in tibial nerve models. CONCLUSION: The strong gene expression in neurons and PNS renders this system particularly attractive for neuroanatomical tracing studies. Furthermore this gene delivery method allowing specific targeting of motor and sensory neurons without damaging the spinal cord might offer potentialities for the gene therapy of peripheral nerve injury. PMID- 15828480 TI - [Fabrication and application in vitro of pedicle guide device for pedicle screws insertion]. AB - OBJECTIVE: To evaluate the accuracy of pedicle guide device for the placement of the pedicle screws. METHODS: Pedicle guide device was designed and made for the anatomical trait of pedicle. The 3-D anatomical data of the thoracic pedicles were measured by multi-slice spiral CT in two embalmed human cadaveric thoracic pedicles spine (T1-T10). Depending on transverse section angle (TSA) and sagittal section angle (SSA) of pedicle axis, the degree of horizontal dial and sagittal dial were adjusted in the guide device. The screws were inserted bilaterally in the thoracic pedicles by using the device. After pulling the screws out, the pathways were filled with contrast media. The TSA and SSA of developed pathways were measured. RESULTS: Analysis of the difference between pedicle axis and developed pathway was of no statistical significance (P > 0.05). CONCLUSION: The guide device could be easily operated and guarantee high accuracy of the pathways of screws and the incidence of pedicle penetration could be significantly reduced. PMID- 15828481 TI - [The clinical effect of anti-rotation reduction internal fixator on the treatment of fresh thoracolumbar spine fracture]. AB - OBJECTIVE: To evaluate the effect of self-designed anti-rotation reduction internal fixator (ARRIF) on treating different spine segment fracture. METHODS: From August 1999 to March 2003, 76 patients(48 males and 28 females, aged from 22 to 59 with an average of 34.1) with thoracolumbar fracture were operatively treated by ARRIF. The follow-up period ranged from 6 to 21 months (15 months in average). Classification according to injury segment: flexion compression fracture 27 cases, burst fracture 42 cases, flexion distraction injury 3 cases, flexion revolving type fracture dislocation 2 cases, shear force type dislocation 2 cases. Classification according Frankel's grade: A grade 16 cases, B grade 15 cases, C grade 27 cases, D grade 10 cases, E grade 8 cases. Operation duration, volume of bleeding, incidence post-operation complication and effect of reduction fixation were observed. RESULTS: The operation duration of ARRIF was 1.2 h in average, and there was about 200 ml volume of bleeding during operation. The nerve function showed one Frankel's grade improvement after operation were as follows: A grade 8 cases (50%), B grade 11 cases (73.3%), C grade 20 cases (74.1%), D grade 3 cases (30%); 2 Frankel's E cases have no nerve function changes. The nerve function damage have no aggravation in all the patients, the postoperation Cobb's angle was averagely corrected 22 degrees. The horizontal displacement of dislocation vertebrae was averagely corrected 28% in sagittal plane, the statistical analysis had significant variance (P < 0.01). ARRIF had no complications of the breakage of screws and rods. CONCLUSION: ARRIF proves to be a valid internal fixator in reducing and fixing different thoracic lumbar segment spine fracture. PMID- 15828482 TI - [The effect of fibroblast growth factors and epidermal growth factors on the proliferation of rabbit anterior cruciate and medial collateral ligament cell in vitro]. AB - OBJECTIVE: To investigate the effect of acid, basic fibroblast, growth factor (aFGF, bFGF) and epidermal growth factor (EGF), and their combination on the proliferation of rabbit anterior cruciate ligament (ACL) and medial collateral ligament (MCL) in vitro. METHODS: The cells of ACL and MCL were isolated and subcultured from the knee joints of ten-week-old New Zealand white rabbits. The cells were seeded into 96-well corning cluster plates. Three growth factors of different concentration alone or in combination were added into the culture medium respectively, which were 0, 1, 5, 10, 50 and 100 ng/ml for aFGF, bFGF and 0, 1.56, 3.13, 6.25, 12.5, 25 and 50 ng/ml for EGF. The proliferation of the fibroblasts was measured for 48 h with XTT method. RESULTS: All of the three growth factors alone promoted the cell proliferation of ACL and MCL fibroblasts. The concentration of aFGF had a significant effect on the proliferation of both ACL and MCL fibroblasts. The concentration of 1 ng/ml bFGF and 5 ng/ ml EGF was most effective in promoting the proliferation of ACL, and both bFGF and EGF had a significant effect on MCL. 5 ng/ml aFGF with 50 ng/ml EGF had effect on ACL. 1 ng/ml aFGF with 3.13 ng/ml EGF had effect on MCL. CONCLUSION: The three growth factors may promote the cell proliferation of ACL and MCL. These findings suggest that topical application of aFGF, either alone or in combination with EGF may have the potential to promote the proliferation of rabbit ACL and MCL,and aFGF of low concentration in combination with EGF is more effective than single growth factor. PMID- 15828483 TI - [Biocompatibility study on tendon mixed extraction of bovine collagen for periodontal tissue engineering]. AB - OBJECTIVE: To study the biocompatibility of tendon mixed extraction of bovine collagen (tMEBC) and to explore the feasibility of using the three-dimensional framework as periodontal tissue engineering scaffold. METHODS: After being prepared, the tMEBC were cultured with the P4-P6 of human periodontal ligament fibroblasts (HPDLFs) in vitro. Three-dimensional framework was prepared from bovine tendon. The P4-P6 of HPDLFs (with an initial density of 5 x 10(6) cells/ml) were cultured in vitro. Cell attachment and proliferation were measured by cell counting 1 day, 3, 5, and 10 days after cell seeding. Histological examination was performed with light microscope and scanning electron microscope 5 and 10 days after cell seeding. RESULTS: Porous structure, which supported the proliferation and attachment of HPDLFs, was found in tMEBC. The density of cell increased from 0.556 x 10(4) cells/ml 24 hours after cell seeding to 3.944 X 10(4) cells/mI 10 days after seeding. Light and scanning electron microscope examination indicated that HPDLFs were attached and extended on the three dimensional scaffolds and were well embedded in the newly formed tissue matrix. CONCLUSION: tMEBC has good biocompatibility with the HPDLFs, and can be used as scaffold for cell transplantation in periodontal tissue engineering. PMID- 15828484 TI - [Culturing and cryopreservation of pre-cartilaginous stem cells from neonate rat]. AB - OBJECTIVE: To establish an effective way to cryopreserve pre-cartilaginous stem cells (PSCs) of neonate rat. METHODS: PSCs [fibroblast growth factor-3 (FGFR-3) positive cells] were isolated and purified by magnetic cell sorting method. PSCs were cultured and amplified to the third generation. PSCs were preserved in liquid nitrogen. The biological properties of cryopreserved PSCs were investigated by reverse transcriptase polymerase chain reaction (RT-PCR), immunohistochemistry, and immunofluorescence. RESULTS: Immunohistochemical and immunofluorescent analysis showed widespread expression of FGFR-3 in cryopreserved PSCs. FGFR-3 could be detected by RT-PCR in cryopreserved PSCs. Cryopreserved PSCs kept high cell viability, and phenotypic and proliferation characteristics of PSCs in vivo. CONCLUSION: Cryopreservation of PSCs can supply adequate qualified cells for repairing the defects of epiphyseal growth plate by tissue engineering technique. PMID- 15828485 TI - [Bio-derived bone material]. AB - OBJECTIVE: To review researches on bone defect repaired by different kinds of bio derived bone. METHODS: The recent relevant literatures were extensively investigated. Preparation of bio-derived bone and effect of bone defect repair were reviewed. RESULTS: The allogeneic and xenogeneic bone treated by different physicochemical method were not only the substitution for bone but also the scaffold material co-cultured with seed cells to reconstruct tissue engineered bone. CONCLUSION: The tissue engineered bio-derived bone is a breakthrough for treatment of bone defect. PMID- 15828486 TI - [Marrow stromal cells and repair of bone defect]. AB - OBJECTIVE: To review researches on the relation between marrow stromal cells (MSCs) and repair of bone defect. METHODS: The latest original literatures about marrow stromal cells and their use in the treatment of bone defect were extensively reviewed. RESULTS: Marrow stromal cells were induced to osteoblasts under proper conditions and showed the potential of bone formation in vivo. The methods of bone tissue engineering using MSCs as seed cells and gene therapy using MSCs as target cells were both useful in the repair of bone defect. CONCLUSION: MSCs have a promising future in the repair of bone defect. PMID- 15828487 TI - [A survey of some new drugs]. PMID- 15828488 TI - [Appropriate use of medicines in patients with rheumatoid arthritis, osteoarthritis and osteoporosis--pharmacists' role]. AB - The care of patients with osteoarthritis, rheumatoid arthritis and osteoporosis often requires a pharmacologic approach. The co-prescription of several medicines is regularly needed in order to optimise treatment. Several recent studies, however, have identified instances of inappropriate medicines use in these patients. This mainly includes "misuse" (inappropriate medicines use in terms of dosing, choice of drug, treatment modalities, treatment duration, interactions,...) and "underuse" (omission of drug therapy that is indicated for the treatment or prevention of a disease or condition). The following examples are discussed: insufficient or inappropriate pain control, use of codeine and tramadol in CYPD2D6 poor metabolisers, underuse or delayed use of disease modifying anti-rheumatic drugs, medication errors with methotrexate, underuse of medicines for the treatment of osteoporosis. By being aware of these events of inappropriate medicines use, the pharmacist should be better able to deliver pharmaceutical care to these patients. The aim is to improve patients' quality of life. PMID- 15828489 TI - [Review of the use of artemisinin and its derivatives in the treatment of malaria]. AB - This article reviews the development of the artemisinins used in the treatment of drug-resistant Plasmodium falciparum malaria. The story starts in China with Artemisia annua L., a plant that was traditionally used as an antipyretic. The activity of Annual wormwood can be explained by the presence of the active substance artemisinin. Soon, artemether, artemotil, artenimol, artesunate and sodium artesunate, derivatives of artemisinin, have been developed. Each has its own physical and pharmaceutical properties, dosage and dosage forms. Other aspects, such as the general guidelines for use, safety during pregnancy and the perspectives of artemisinin compounds, are being discussed. PMID- 15828490 TI - [Osteoporosis in 2005: diagnose and treatment]. PMID- 15828491 TI - [Paracelsian drug compounding in the 16th and 17th century]. PMID- 15828492 TI - Mechanical signalling: lessons from the past that control the future. AB - Relative movement between an implanted material and tissues can play a decisive role in generating the host response. As with other components of the biocompatibility of devices, it is essential to control the mechanical signalling associated with this movement if we are to optimise this response. PMID- 15828493 TI - Programmable elution profile coating for drug eluting stents. AB - Despite good results in reducing in-stent restenosis during cardiac stenting procedures, a number of issues in the application of drug-eluting stents still challenge the industry. These include drug-elution kinetics and long-term biocompatibility. Some of the latest developments in these areas are described here. PMID- 15828494 TI - Benefits of the rotary diaphragm pump. AB - The huge variety of applications in the medical field represents a challenge for the design of miniature pumps. There are well-known designs such as piston pumps, eccenter diaphragm pumps and peristaltic pumps. There are lesser-known types such as the rotary diaphragm pump, the subject of this article. Its design features, variants, and advantages and disadvantages are examined. PMID- 15828495 TI - US and European postmarket clinical data requirements. AB - Under United States regulations, the Food and Drug Administration may require manufacturer to collect postmarket clinical data. In Europe, the role of postmarket clinical follow-up is described in a guidance document, but not specifically mentioned in the European Directives. This article discusses the two systems, which are similar, but also different in important aspects. PMID- 15828496 TI - Avoiding loss of medical information during clinical investigations. AB - One of the challenges in conducting clinical investigations of medical devices is obtaining reliable data. This article outlines some issues to consider to help achieve this. PMID- 15828497 TI - Marriage guidance for collaborators. AB - Collaboration with external partners is a way to accelerate product innovation. Tips on building successful partnerships are offered here. PMID- 15828498 TI - Changing emphasis in test for patent infringement. AB - The United Kingdom is moving away from its current method for assessing infringement of a patent. PMID- 15828499 TI - New medical device perspectives from integrated optics and fibre optics. AB - Integrated optics technology has a great potential for medical device applications. This article explains the technology and how it can be used with particular reference to in vitro diagnostics and endoscopy. PMID- 15828500 TI - A review of plastic welding processes. PMID- 15828501 TI - Multi-strategies to implement health promoting schools. PMID- 15828502 TI - The concept of health promoting schools to enhance positive youth development. AB - The concept of Health Promoting Schools improves the school's physical and social environment, its curriculum, teaching and learning methods, and the personal and social development of students. It will enable the participating schools create a 'learning perspective', 'community perspective', and 'capacity building' environment to improve both educational and health outcomes of students. As the scheme promotes self-esteem of students, staff development, parental education, involvement of the whole school community and linkage with different stakeholders, it will also enhance positive youth development. It will help to shift the paradigm from handling crisis to prevention approaches emphasizing on youth support before problem behaviours occur, and also address the circumstances (families, schools, communities, peer groups) of youth's lives. PMID- 15828503 TI - The importance of needs assessment in planning health promoting schools initiatives: comparison of youth risk behaviours of two districts in Hong Kong. AB - A needs assessment is recommended to be carried out before planning resource allocation in a community as it could help to identify the needs of the people. This study illustrated the importance of needs assessment in planning health promoting school initiatives by comparing the results of youth risk behaviours surveys conducted at two districts of Hong Kong, namely Tsuen Wan and North District. The findings indicated that the two districts should prioritize their resources in respect to their urgent needs. A higher proportion of students from Tsuen Wan participated in vigorous exercise regularly in comparison to students from North District. Students from North District consumed more vegetable per week and exhibited less depressive symptoms than students from Tsuen Wan. The availability of data on youth risk behaviours of the two districts would provide information for strategic planning and direct decision-making in youth health programmes. PMID- 15828504 TI - Significance of the Hygiene Charter towards different sectors in Hong Kong. AB - The occurrence of SARS in March 2003 has resulted in an increased interest, worldwide in emerging infectious diseases. The SARS experience provided us a lesson on the importance of promoting hygienic practices among individuals and different working sectors. In Hong Kong, a voluntary organization called the UNITE proposed a Hygiene Charter which aimed at taking hygiene to new levels. This action has been supported by individuals and different sectors including the Personal and Family, Management, Buildings, Catering, Education, Finance and Commercial, Industrial, Medical and Health, Public Transportation, Social Welfare, Sports and Culture and Tourism. As promotion and maintenance of environmental health requires input from different sectors, the signing of the Hygiene Charter provides an opportunity for individuals and the public to show their pledge and commitment to good hygiene practices. As a result, with environment improvement and good infectious disease control measures, prevention of epidemics of infectious diseases is deemed to be possible. PMID- 15828505 TI - What are the needs of students? An experience from a district based health promoting schools project in Hong Kong. AB - A district based needs assessment as conducted over the past year to uderstand the health problems and the health education needs of the students in Tai Po district, Hong ong. 6879 primary one to primary six students from eighteen rimary schools, participating in a district based Health Promoting Schools Project in Hong Kong, were invited to complete a self-administrated questionnaire which was partly adapted from the Centre for Diseases and Control (CDC)'s Youth Risk Behaviour Surveillance Survey. The results presented a holistic picture of the health and needs of primary students with respect to their general health status, mental health, body weight and dietary behaviour, exercise, preventive health care, tobacco use, alcohol and other drug use, school environment and school health education. It provides baseline information for the project to prioritize the problems and strategically plan health promotion 'programmes with reference to the concept of Health Promoting Schools by the World Health Organization. PMID- 15828506 TI - Parent training: experience of the New Territories West School Health Promotion Project of Hong Kong. AB - Parents' participation in school life is an important element of a health promoting school. To maximize the potential of parents as partners in health education and take on a leading role in promoting health in the school, family and community, a parental health education programme using the empowerment model had been launched in partnership between academic and health sectors. A total of 28 parents selected from eight schools in the New Territories West region of Hong Kong participated in the programme. Evaluation of the programme revealed that the programme had matched well with the expectation of most participants. All respondents had reported an increase in health awareness and knowledge, and confidence to promote health concepts in familiar environments, such as the home and school. They also showed interest to participate in further training in health related issues. Parental health education is recommended to enhance active involvement for building a greater sense of belonging and to put through individual empowerment to community empowerment. Parental involvement in school health promotion would be an effective way to facilitate the paradigm shift. PMID- 15828507 TI - To be a responsible adolescent: a key strategy to enhance students' awareness of self-care and their health behaviours. AB - This article presents unique teacher's experience in developing and implementing a highly acclaimed pioneer comprehensive health-promoting programme at a secondary school in Hong Kong. The authors explain why a systematic school health education is needed from both personal and public health perspectives. A five year programme known as 'Be responsible' had been introduced at St. Catharine's School for Girls since 1999 with an objective of guiding students to assume responsibility for their health. Through interactive activities, students had learnt and acquired the knowledge, skills and attitude that would empower them to enjoy an all-rounded development and life-long well-being. The Programme adopted a 'bottom-up' model that extended its benefits beyond the classrooms. Health education follows the modern education theory of holistic development and is the foundation of a quality education system. Teachers should therefore be committed to the cause and role of promoting health at school and work in partnership with the health professionals. PMID- 15828508 TI - The story of a school participating in the Healthy School Award Scheme in Hong Kong. AB - This article described why the Tsung Tsin College joined in the Hong Kong Healthy Schools Award Scheme, how it tried to live up to the expectations, and the gains it reaped from the experience. Operating in a less affluent district of Hong Kong, the school strived for whole-person education for its students, many of whom suffer from social deprivations, and turned its environment into one conducive to learning. Finding that the Scheme advocates similar themes, the school enrolled and hoped to strengthen its self-improvement capacity. The school won a Silver Award for its overall performance in health promotion. The audit it underwent has also revealed its weaknesses, which it is striving to remedy. On the other hand, the school has also acquired many benefits from the process. The school-community and even school-parent relationships have also been enhanced as a result of its participation in the Scheme. PMID- 15828509 TI - How can we build resilience in primary school aged children? The importance of social support from adults and peers in family, school and community settings. AB - This study examines the association between firstly, student resiliency and their perceptions of social support from parents/caregivers, teachers, and peers, and secondly, between student's perception of their general health status and their social support. A cross-sectional research project was designed and conducted in 2003 in an urban and remote area of Queensland, Australia. The study population comprised of 2580 students (Years three, five, and seven) across 20 primary schools. The main outcome measures were self-reported health status and resiliency behaviours. Independent variables included student perceptions of support from parents/caregivers, teachers, school peers, and prosocial groups. Students who perceived parents, teachers, and peers as supportive were more likely to have higher resiliency behaviour in communication and cooperation, self esteem, empathy, help-seeking, goals and aspirations. Students who considered that their parents, peers at school and prosocial groups were supportive, were more likely to feel healthy. Findings suggest that providing adult and peer support to students at primary school age is a vital strategy in promoting student resiliency and general health for children of primary school age. PMID- 15828510 TI - The health promoting school and social justice in a global environment. AB - Globalisation is present whether recognised in SARS, global terrorism, finance or youth music. With the growth of the health promoting school movement in this context and the increased numbers of countries and schools involved, eight themes are proposed as critical to how the Health Promotion School move forward. They are concerned with: the diverse origins and alliances of forces in the movement; holistic and ecological approach; its status as a global movement; the tension between and empowerment or compliances model; evidence-based and values-based approches; the radical vision; social capital and social inclusion; and sustainability. Reaching the level of acceptance the Health Promotion School has acheived may lead to settling into comfortable official recognition--and assured funding--and losing its militancy. Can the Health Promotion School challenge health inequalities on a national and international scale and can it be a force for social inclusion? PMID- 15828511 TI - Direct access. PMID- 15828512 TI - Using optical coherence tomography to guide articular cartilage ablation. AB - Current clinical imaging technologies are not capable of accurately resolving the microscopic components of articular cartilage. Optical coherence tomography (OCT) is a recently developed imaging modality analogous to ultrasound--OCT measures backreflection of infrared light instead of sound. In the study reported here, we wanted to determine the efficacy of OCT for monitoring cartilage laser ablation. Real-time imaging was conducted on bovine cartilage ablated by an argon laser in vitro. Image sequences were generated illustrating the extent of tissue degradation postablation. The images accurately correlated with histology. These results indicate that current ablation procedures could benefit from OCT guidance. PMID- 15828513 TI - Giant cell tumor of tendon sheath. AB - Neoplasms of the hand are not common. Giant cell tumor of tendon sheath (GCTTS) is the most common primary tumor of the hand. Many different theories have recently been proposed as to whether GCTTS is a neoplasm or a localized reactive process. We believe the evidence supports a neoplastic origin. Although the origin is still not proved, the presentation, diagnosis, and treatment of GCTTS have been clear for a long time. PMID- 15828515 TI - Cryptococcal arthritis of the knee in an immunocompetent host. PMID- 15828514 TI - Outcome of surgery for lateral epicondylitis (tennis elbow): effect of worker's compensation. AB - We retrospectively compared the results of extensor origin release for lateral epicondylitis (tennis elbow) against worker's compensation (WC) status. All patients (N = 57) underwent extensor origin release between October 1989 and June 1998. For the 33 patients (37 elbows) who received WC, mean follow-up was 55 months; for the 25 patients (26 elbows) who did not receive WC, mean follow-up was 45 months. Pain relief, symptom recurrence, satisfaction with procedure outcome, and ability to return to work (same or similar job) were evaluated. Pain relief was reliably achieved in the WC and non-WC groups (36/37 and 24/26 elbows, respectively). Symptom recurrence was intermittent in both groups, and few patients sought medical intervention for recurrent symptoms. Patient satisfaction was high in the WC and non-WC groups (35/37 and 26/26 elbows, respectively). A majority of patients in both groups returned to work, but a significantly higher percentage of patients in the WC group changed jobs because of persistent symptoms. PMID- 15828516 TI - Synovial chondromatosis in the pediatric patient. PMID- 15828517 TI - Spontaneous resolution of a locked long metacarpophalangeal joint: a case report and review of treatment. PMID- 15828518 TI - Ochronotic arthropathy: a report of 3 cases. PMID- 15828519 TI - Volar plating of dorsally comminuted fractures of the distal radius: a 6-year study. AB - This article describes a 6-year study of use of volar plating in treating unstable dorsally comminuted distal radius fractures. Forty-six fractures treated with volar plating were reviewed. All cases were followed for at least 6 months; 35 were followed for more than 1 year. Fractures treated with adjunctive protruding pins were excluded. Bone graft was used in 4 cases. Anatomical results were rated excellent or good in 39 cases (85%). Subsidence occurred in 9 fractures, 4 of which malunited. Flexor pollicis longus tendon attrition occurred in 2 cases. Mean operative time was less than 1 hour (range, 30-90 minutes). Study results support continued use of volar plating in treating selected unstable dorsally displaced fractures of the distal radius. Prominent plates and sharp-edged screws may imperil deep flexor tendons and should be removed. More liberal use of bone graft and improvements in plate design should reduce complications. PMID- 15828520 TI - Supra-acetabular placement of external fixator pins: a safe and expedient method of providing the injured pelvis with stability. AB - Applying a stable anterior pelvic external fixator frame is a skill that should be mastered by all orthopedic surgeons who treat acutely injured patients. Splinting of an unstable pelvis during resuscitation can help to reduce the volume of the true pelvis, pending definitive surgical stabilization of the pelvic ring. Supra-acetabular pin placement, less familiar to most surgeons than iliac wing pin placement is, can provide a more reliable pin-bone interface and thus allow improved reduction ability with fewer soft-tissue complications. Because of their location, supra-acetabular pins also seem to be better tolerated than iliac crest pins when used for definitive management of the pelvic ring disruption. A young man who sustained a type II anteroposterior compression injury in a motor vehicle accident presented with symphyseal disruption (7 cm wide) and left anterior sacroiliac joint disruption. During resuscitation, the pelvis was anatomically reduced and stabilized with a supra-acetabular pin-based external fixator. Pin locations, chosen using palpable and cutaneous landmarks, were inserted without additional imaging guidance. The fracture was reduced anatomically, and the frame was used for definitive management of the pelvic ring injury. PMID- 15828521 TI - Reframing research on sexual behavior and HIV. AB - This review of recent studies examines the conceptual and methodological tools that are used to understand sexual behavior and HIV. Whereas surveys have provided reliable indicators of behavior, their validity is uncertain, their correlation with biological indicators tenuous, and the explanations they offer limited. Microlevel research has called into question the rationalizing, medically informed, and risk-averse frameworks used in the public health literature on HIV and behavior, but its impact has been limited. Thus, surveys and qualitative research have proceeded along two separate tracks, and this separation has hampered our understanding of sexual behavior. A small but growing number of studies, however, provide examples of less-compartmentalized research that avoids oversimplification. They show that expanding the models that drive research in this area is possible, and that efforts to combine disciplinary perspectives result in deeper understandings of knowledge, risk perceptions, attitudes, and behavioral change. PMID- 15828522 TI - Hormonal contraception and physiology: a research-based theory of discontinuation due to side effects. AB - Side effects influence the acceptability and continuation of hormonal contraceptives. Counseling the client about the management of side effects is a principal approach advocated for increasing continuation. Evidence of a biological basis for variation in women's tolerance of hormonal contraceptives argues, however, that greater attention should be given to altering the product rather than principally attempting to alter a woman's ability to deal with the product. Discontinuation rates for hormonal contraceptives, largely attributable to side effects and health concerns, are high in nearly all less-developed countries for which Demographic and Health Survey data are available. Oral contraceptives appear to be particularly problematic for Latin American women, most notably in Bolivia. Clinical trials suggest substantial variation in the physiological response to exogenous hormones, and new evidence confirms the hypothesis that the normal hormonal profiles of Bolivian women are significantly lower than those of women in the United States. These findings suggest a need for more population-specific physiological research linked to analyses of the possible association between endogenous hormone differences and contraceptive continuation. Appropriately adjusting the level of the steroid delivered may benefit women's health and improve the acceptability and continuation of hormonal contraceptives. PMID- 15828523 TI - The impact of franchised family planning clinics in poor urban areas of Pakistan. AB - This study uses a quasi-experimental design to determine the impact of new family planning clinics on knowledge, contraceptive use, and unmet need for family planning among married women in poor urban areas of six secondary cities of Pakistan. Baseline (n = 5,338) and endline (n = 5,502) population surveys were conducted during 1999-2000 and 2001-02 in four study sites and two control sites. Exit interviews with clients identified the sociodemographic and geographic characteristics of clinic users. The results show that the clinics contributed to a 5 percent increase in overall knowledge of family planning methods and an increase in knowledge of female sterilization and the IUD of 15 percent and 7 percent, respectively. Distinct effects were found on contraceptive uptake, including an 8 percent increase in female sterilization and a 7 percent decline in condom use. Unmet need for family planning declined in two sites, whereas impacts on the other sites were variable. Although the new clinics are located within poor urban communities, users of the services were not the urban poor, but rather were select subgroups of the local population. PMID- 15828524 TI - Use of health professionals for obstetric care in northern Ghana. AB - This study explores the role of access versus traditional beliefs in the decision to seek obstetric care from health professionals. Eighteen purposively sampled homogenous groups in Kassena-Nankana District of northern Ghana participated in focus-group discussions about traditional beliefs, barriers to the use of health professionals, and ways to improve obstetric care. All the groups were knowledgeable about the life-threatening signs and symptoms of complications of pregnancy and labor. Decisions about place of delivery generally were made after the onset of labor. Accessibility factors (cost, distance, transport, availability of health facilities, and nurses' attitudes) were major barriers, whereas traditional beliefs were reported as less significant. Informants made pertinent recommendations on how to improve obstetric services in the district. These findings demonstrate that even in this district, where African traditional religion is practiced by a third of the population, compared with a national average of 4 percent, lack of access was perceived as the main barrier to seeking professional obstetric care. PMID- 15828525 TI - Factors influencing boys' age at first intercourse and condom use in the Shantytowns of Recife, Brazil. AB - Despite the general recognition that the sexual practices of adolescent boys place them at high risk of acquiring sexually transmitted infections (STIs), including HIV, and of causing unwanted pregnancies, advances in mapping their sexual behaviors have been slow. This study uses data recently collected from low income areas of the city of Recife, Brazil, to study boys' age at first sexual intercourse and factors that hinder their use of condoms. These boys become sexually active at early ages, and despite their general awareness of HIV, they rarely use condoms, especially at ages younger than 15. Sustained family involvement in guiding boys is associated with later first intercourse and an increased use of condoms. Boys who describe themselves as shy with girls have later first intercourse, although the probability of their using condoms does not differ from that of other boys of their age. Higher socioeconomic status leads to earlier sexual activity for boys (in contrast with girls), but also to a greater likelihood of using condoms during first intercourse. PMID- 15828526 TI - Reproductive health and the Millennium Development Goals: the missing link. AB - This report was commissioned by the Population Program of the William and Flora Hewlett Foundation in December 2004. The author was charged with analyzing the United Nations' deliberations that led to the adoption of the Millennium Development Goals (MDGs) to answer the question of why there is no specific reproductive health goal. Her report, of which this is a lightly edited version, is also available online at . This coverage of the MDG process will be complemented by a special section of Studies in the June 2005 issue on reproductive health and the MDGs. The section will include excerpts about reproductive health from the final report of the Millennium Development Project entitled "Investing in Development: A Practical Plan to Achieve the Millennium Development Goals," which was submitted to Secretary General Kofi Annan in January 2005, as well as commentaries by leading scholars and policymakers in the fields of population and reproductive health. PMID- 15828527 TI - Eritrea 2002: results from the Demographic and Health Survey. PMID- 15828528 TI - Indonesia 2002-2003: results from the Demographic and Health Survey. PMID- 15828529 TI - Every unit is unique. PMID- 15828530 TI - Making connections. PMID- 15828532 TI - Management. Scanning the globe. AB - Overseas "nighthawks" can make life easier for radiologists by reading and interpreting scans and X-rays during your staff's off hours. The growing practice does have its critics, however. PMID- 15828533 TI - Patient care. Reporting errors. PMID- 15828534 TI - Awards. HRET honors expert. PMID- 15828535 TI - Staffing. Turbo-charging CIOs. AB - At CHIME's CIO boot camp, seasoned chief information officers give their newer colleagues an intensive, three-day "immersion" in the new realities of their profession. PMID- 15828536 TI - Technology. Rural discount. AB - The Federal Communications Commission recently changed its definition of rural community, which could mean more affordable telecommunications rates for certain providers. June 30 is the deadline for funding applications. PMID- 15828537 TI - Patient care. Giving kids a voice. PMID- 15828538 TI - Facilities. Get out the China. PMID- 15828539 TI - Data page. The "under-65" uninsured. PMID- 15828540 TI - Three companies to watch. PMID- 15828541 TI - What nurses want. Different generations. Different expectations. AB - Younger nurses have different career expectations than their older colleagues. They seek a balanced lifestyle with reasonable work hours, demand to use the latest technology and expect to be vocal members of the care team. If a hospital disappoints, they are quick to change jobs. How do you accommodate them without alienating veteran staff members? PMID- 15828542 TI - Even you can control energy costs [a bit]. AB - Senior executives pay relatively scant attention to their hospital's utility bills. They are a relatively small portion of overall costs, and anyway, executives reason, not much can be done about them. But innovative energy managers dispute such thinking. They're attacking the problem from entirely new angles, and the savings for hospitals can be significant. PMID- 15828543 TI - Experimenting with Medicaid. AB - Big budget deficits and soaring health care costs have states scrambling to reduce Medicaid spending. Not every governor is choosing to simply cut the program, however; some are experimenting with new ways to meet the needs of their poor residents without bankrupting their states. The federal government and others are watching closely. PMID- 15828545 TI - Hospitals must improve workplace communication to help reduce medical errors. PMID- 15828544 TI - The supply chain and clinical quality. AB - Improving quality of care has become a major consideration when it comes to buying supplies. Here's how leading-edge hospitals are changing the process, including adding clinicians to their buying staffs and standardizing how they choose and use products. PMID- 15828546 TI - Where to start? PMID- 15828547 TI - Letrozole improves disease-free survival vs tamoxifen in adjuvant treatment of early breast cancer. PMID- 15828548 TI - A postmenopausal woman with early-stage breast cancer and borderline hormone receptor staining. PMID- 15828549 TI - The horizon of antiangiogenic therapy for colorectal cancer. AB - Vascular endothelial growth factor (VEGF) plays a crucial role in the growth and metastatic spread of cancer. Bevacizumab (Avastin) is the first commercially available VEGF inhibitor, earning U.S. Food and Drug Administration (FDA) approval in February 2004. In combination with fluorouracil (5-FU)-based chemotherapy, this agent significantly prolongs overall and progression-free survival of patients with metastatic colorectal cancer. This review details the emerging role of the drug, its unique side effects, and other practical considerations related to bevacizumab therapy. Ongoing trials attempting to define additional indications for bevacizumab as well as the development of other promising angiogenesis inhibitors are also reviewed. PMID- 15828550 TI - Organ preservation in muscle-invasive bladder cancer. AB - While organ preservation with nonextirpative surgery, radiotherapy, and frequently, chemotherapy has become a favored strategy in the treatment of many cancers, bladder preservation for patients with invasive disease remains controversial. The standard treatment for muscle-invasive bladder cancer in the United States is still radical cystectomy with pelvic lymph node dissection. An alternative to cystectomy is multimodality bladder preservation with thorough transurethral resection, chemotherapy, and radiation therapy. This review will address issues raised by a multimodality approach for the treatment of invasive bladder cancer. PMID- 15828551 TI - Clinical trials referral resource. Current clinical trials investigating 3-AP. PMID- 15828552 TI - Nutritional support of patients undergoing radiation therapy for head and neck cancer. AB - Malnutrition plays a key role in the morbidity of head and neck cancer patients receiving surgery, chemotherapy, radiotherapy, or combined-modality therapy. In addition to weight lost prior to the diagnosis of head and neck cancer, the patient may lose an additional 10% of pretherapy body weight during radiotherapy or combined-modality treatment. A reduction of greater than 20% of total body weight results in an increase in toxicity and mortality. Severe toxicity can result in prolonged treatment time, which has been implicated in poor clinical outcome. Early intervention with nutritional supplementation can reduce the chance of inferior outcome in patients at high risk of weight loss. The preferred route of nutritional support for these patients is enteral nutrition. Two commonly used methods for enteral feedings are nasoenteric and percutaneous endoscopic gastrostomy. It is important to take into account the ethical considerations involved in providing long-term nutritional support, particularly for patients with terminal conditions. Nutritional directives are best evaluated through multidisciplinary efforts, including input from the patient as well as members of the nursing, nutritionist, and medical staff. PMID- 15828553 TI - Zoledronic acid prevents cancer treatment-induced bone loss. PMID- 15828554 TI - Combined-modality treatment for operable pancreatic adenocarcinoma. AB - Although in centers where pancreatectomy is performed frequently, associated morbidity and mortality rates have improved, long-term outcomes in pancreatic adenocarcinoma patients remain poor when surgery is the sole therapeutic modality. The impact of adjuvant chemotherapy on survival in patients with localized pancreatic cancer remains incompletely defined. The European Study Group for Pancreatic Cancer (ESPAC)-1 trial has suggested that overall survival rates are superior when chemotherapy is added to surgery, even when regimens believed to be relatively ineffective in the treatment of advanced disease are used. The role of radiotherapy given with chemotherapy is also unresolved, but chemoradiation continues to receive consideration in the multimodality approach to localized pancreatic cancer. Enhanced collaboration and increased involvement by pancreatic surgeons have helped in the recruitment of pancreatic cancer patients for large-scale randomized clinical trials in Europe and the United States. Many newer chemotherapeutic agents with efficacy in gastrointestinal cancers have yet to be investigated in the adjuvant and neoadjuvant settings. PMID- 15828555 TI - JCAHO performance pay guidelines hard to meet. PMID- 15828557 TI - Better facility design leads to better health care outcomes. PMID- 15828556 TI - Anti-trust suits should be against HMOs. PMID- 15828558 TI - Hospital wait list lessons from the UK. PMID- 15828560 TI - Re-inventing the microscope: the Canadian Light Source (CLS). PMID- 15828559 TI - Initial learnings as the OHAfrica team commences its work at an HIV clinic in Lesotho. PMID- 15828561 TI - The changing landscape for COX-2 inhibitors: a summary of recent events. PMID- 15828562 TI - Building capacity in applied health services research. PMID- 15828563 TI - More scans, more scanners. PMID- 15828564 TI - The patient safety journey. AB - The patient safety journey will involve front line providers, researchers, patients, community and institution-based care teams and others, and many changes in the way organizations and procedures are structured. This is no small task, yet is critical: the Canadian public and healthcare providers are rightly asking that real strides be made. "At the end of our first five-year mandate, I hope that we can point to the results of a new Canadian Adverse Events study and say, 'Yes, the Canadian health system is safer than it was when we started and it will continue to get safer as we learn more,'" said Dr. John Wade. "That's what success should look like." PMID- 15828565 TI - Healthcare renewal in Canada: accelerating change. PMID- 15828566 TI - Making hospital care safer and better: the structure-process connection leading to adverse events. AB - The 2004 Canadian Adverse Events (AE) Study which focused on estimating the incidence of AEs in hospital settings found an annual AE rate of 7.5% of all hospital admissions in Canada (Baker et al. 2004). Although study findings in terms of patient outcomes are essential for hospitals to review their own practices, a thorough understanding of the contributing factors leading to adverse events will be an important next step for hospitals to correct the deficiencies that might act as barriers to providing safe patient care. In other words, a thorough investigation and analysis of structure and process factors within individual hospitals as they relate to patient outcomes is needed as a next step. In order to explore how structural and care-process factors might be linked to the occurrence of adverse events in hospitals, this article aims at building a predictive model, by using Donabedian's model of structure, process, outcome and analyzing secondary data of Ontario Registered Nurse Survey of Hospital Characteristics. Results revealed that perceived understaffing, inadequate support services, unpleasant work environment, poor teamwork and non supportive administration impact negatively the number of tasks left undone by nurses, which in turn, influence negatively the occurrence and frequency of adverse events in hospitals. Results and implications are discussed. PMID- 15828567 TI - Urban outpatient views on quality and safety in primary care. AB - OBJECTIVE: The Minimizing Errors Maximizing Outcomes Study is designed to examine the effect of workplace conditions on quality of care and medical errors. In the first phase of the study, patients were asked to "tell their stories" via focus groups. DESIGN: Moderators used a standard question guide. Researchers read the transcripts independently and reached consensus on major themes. Two coders independently assigned transcript statement to themes. SETTING: Three focus groups were conducted in three cities, including 21 patients from three clinics. PATIENTS: Patients with previously scheduled appointments at participating clinics were invited to join the focus groups. MEASUREMENTS AND MAIN RESULTS: Agreement between the two coders was 77.5% (kappa value 0.66). All but 2% of 187 distinct comments could be grouped into four categories: (1) Systems Issues (44% of comments). Long waits for providers and lack of access were the most common frustrations. Understaffing, underfunding and lack of health insurance were perceived as contributing to poor quality of care; (2) Interpersonal Skills (37%). Physician listening skills were valued. Participants felt patient attitudes affected care. (3) Knowledge and Technical Skills (9%). (4) Errors (7%). Medication errors, errors of inattention and technical errors were discussed. CONCLUSIONS: Patients provide important insights into complex systems issues, which can guide planners in improving quality and reducing errors. According to focus group participants, healthcare could be improved and made safer by increasing timely access to patients' own physicians, decreasing the time patients spend in waiting rooms, and adding staff to double-check prescriptions. PMID- 15828568 TI - Evidence, economics and ethics: resource allocation in health services organizations. AB - Priority setting is a challenge for health services organizations across health systems. Because demand for health services exceeds available resources, health services priorities must be set to ensure resources are used appropriately to meet the community's health needs. Various approaches have been developed to assist decision-makers to set priorities in their organizations. The dominant approaches come from evidence-based medicine, economics and ethics. PMID- 15828569 TI - Improving hospital performance: culture change is not the answer. AB - An inappropriate culture is often presented as the reason why hospitals throughout the world have been unable to achieve best practice. Many have concluded that the organizational culture of hospitals limits the ability of these organizations to improve performance, particularly in relation to improving quality and safety. Establishment of a "better" culture is often presented as the resolution to quality, safety, financial and productivity issues in hospitals. Our research indicates that certain management conditions are required before culture change can be contemplated. This paper suggests that we have underestimated the importance of people management in improving practice within hospitals, with the three most important aspects being the development of teamwork, performance management and sophisticated training. We present evidence of the potential contribution good people management can make to high performance, and argue that better people management is a cause, not an outcome, of cultural change. PMID- 15828570 TI - Culture is the outcome of how people interact. PMID- 15828571 TI - Changing roles for primary-care physicians: addressing challenges and opportunities. AB - Direct-to-consumer advertising is but one example of a process called disintermediation that is directly affecting primary-care physicians and their patients. This paper examines the trends and the actors involved in disintermediation, which threatens the traditional patient-physician relationship. The paper outlines the social forces behind these threats and illustrates the resulting challenges and opportunities. A rationale and strategies are presented to rebuild, maintain and strengthen the patient physician relationship in an era of growing disintermediation and anticipated advancements in cost-effective office-based information systems. Primary care--as we know it--is under siege from a number of trends in healthcare delivery, resulting in loss of physician autonomy, disrupted continuity of care and potential erosion of professional values (Rastegar 2004; Future of Family Medicine Project Leadership Committee 2004). The halcyon days of medicine as a craft guild with a monopoly on (1) technical knowledge and (2) the means of implementation, reached its zenith in the mid-twentieth century and has been under pressure ever since (Starr 1982; Schlesinger 2002). While this is a trend within the US health system, it is likely to affect other delivery systems in the years ahead. PMID- 15828572 TI - Primary-care: front, centre and middle. PMID- 15828573 TI - Clinical information systems: understanding and preventing their premature demise. PMID- 15828574 TI - Medication administration 2004. FOCUS: medication administration application performance. PMID- 15828575 TI - Characterization of protective immunity induced against Schistosoma mansoni via DNA priming with the large subunit of calpain (Sm-p80) in the presence of genetic adjuvants. AB - Despite advances in control via snail eradication and large-scale chemotherapy using praziquental, schistosomiasis continues to spread to new geographic areas particularly in sub-Saharan Africa. Presently, there is no vaccine for controlling this disease. We have concentrated on a functionally important schistosome antigen Sm-p80 as a possible vaccine candidate for schistosomiasis. Here we report the proliferation of spleen cells in response to the recombinant Sm-p80 protein and cytokine (IFN-gamma and IL-4) production by the splenocytes. These spleen cells were obtained from groups of mice that were vaccinated with a DNA vaccine formulation containing Sm-p80 and one of the Th-1 (IL-2 or IL-12) or Th-2 (GM-CSF, IL-4) enhancer cytokines. The splenocytes from the groups of mice vaccinated with Sm-p80 DNA in the presence of Th-2 enhancer cytokines showed moderate but detectable proliferation. The splenocytes obtained from mice vaccinated with Sm-p80 DNA with Th-1 enhancer cytokines IL-2 and IL-12 provided the highest proliferation. The IFN-gamma production by splenocytes was found to follow the similar pattern [(Sm-p80) < (Sm-p80 + IL-4) < (Sm-p80 + GMCSF) < (Sm p80 + IL-12) < (Sm-p80 + IL-2)], as has been observed for the proliferation and protection data. However, the elevated IL-4 production was inversely correlated to Sm-p80-induced splenocyte proliferation or the protection. These results show again that protective immune response induced by Sm-p80 is of Th-1 type. PMID- 15828576 TI - Two new species of Eimeria (Apicomplexa: Eimeriidae) from Asian geoemydid turtles Kachuga tentoria and Melanochelys trijuga (Testudines: Geoemydidae). AB - Coprological examination of ten Pink-ringed tent turtles Kachuga tentoria circumdata, recently imported from India, and three Burmese black turtles Melanochelys trijuga edeniana, imported from Myanmar, revealed the presence of two new species of Eimeria. Oocysts of Eimeria kachua n. sp. from K. t. circumdata are broadly oval to subspherical, 15.3 (13-18) x 13.9 (12-16) microm, with polar granule and subspherical oocyst residuum. Sporocyst elongatelly oval to spindle-shaped, 8.7 (7.5-10) x 4.9 (4-6) microm, with a knoblike Stieda body, covered with fine membranous cupola-like structures. Thin walled oocysts of Eimeria patta n. sp. from M. t. edeniana, have an irregular shape, influenced by the position of sporocysts, frequently with lobular irregularities, 12.6 (11-16) x 9.1 (7.5-12) microm. Sporocysts are oval to ellipsoidal, 5.8 (5-7) x 4.2 (3.5 5) microm. PMID- 15828577 TI - Taxonomic reports of Homeacanthoidea (Eucestoda: Trypanorhyncha) in Lamnid and Sphyrnid elasmobranchs collected off the coast of Santa Catarina, Brazil. AB - Elasmobranch specimens of lamnid and sphyrnid captured in 1999 in the State of Santa Catarina, Brazil, were parasitized with homeacanthoid trypanorhynch cestodes: Isurus oxyrinchus Rafinesque, 1810 with Nybelinia lingualis (Cuvier, 1817) Dollfus, 1929; Sphyrna zygaena (Linnaeus, 1758) with Heteronybelinia rougetcampanae (Dollfus, 1960) Palm, 1999. New details of internal morphology and/or scolex and/or proglottid surface ultrastructure are given. Adults of N. lingualis are reported for the first time in the Brazilian coast. PMID- 15828578 TI - Monogeneans from Pangasiidae (Siluriformes) in Southeast Asia: VIII. Four new species of Thaparocleidus Jain, 1952 (Ancylodiscoididae) from Pangasius polyuranodon and P. elongatus. AB - The examination of gill parasites from Pangasius polyuranodon Bleeker, 1852 and P. elongatus Pouyaud, Gustiano & Teugels, 2002 (Siluriformes, Pangasiidae) in Southeast Asia revealed the presence of four new species of Monogenea all belonging to Thaparocleidus Jain, 1952 (Monogenea, Ancylodiscoididae) as defined by Lim (1996) and Lim et al. (2001): T. furcus n. sp., T. infundibulus n. sp., T. sudartoi n. sp. and T. turbinatio n. sp. PMID- 15828579 TI - Second intermediate host land snails and definitive host animals of Brachylaima cribbi in southern Australia. AB - This study of infection of southern Australian land snails with Brachylaima cribbi metacercariae has shown that all commonly encountered native and introduced snails are susceptible second intermediate hosts. The range of infected snails is extensive with metacercariae-infected snails being present in all districts across southern Australia. C. virgata has the highest average natural metacercarial infection intensity of 6.1 metacercariae per infected snail. The susceptibility of birds, mammals and reptiles to B. cribbi infection was studied in South Australia by capturing, dissecting and examining the intestinal tract contents of animals which commonly eat land snails as a food source. Indigenous Australian little ravens (Corvus mellori), which are a common scavenger bird, and two other passeriform birds, the black bird (Turdus merula) and the starling (Sturnus vulgaris), which are both introduced European birds, were found to have the highest infection rates of all animals examined. Other birds found infected with B. cribbi were an emu (Dromaius novaehollandiae), chickens (Gallus gallus) and a pigeon (Columba livia). Natural infections were also detected in field mice (Mus domesticus) and shingleback lizards (Tiliqua rugosa) although the intensity of infection was lower than that observed in birds. Susceptibility studies of laboratory mice, rats and ducks showed that mice developed patent infections which persisted for several weeks, rats developed a short-lived infection of three weeks' duration and ducks did not support infection. This study has shown for the first time that a brachylaimid can infect a wide host range of birds, mammals and reptiles in nature. PMID- 15828580 TI - [Two new fleas (Siphonaptera: Rhopalopsyllidae: Parapsyllinae) of Argentina, from Abrocoma uspallata Braun and Mares, 2002, (Rodentia: Abrocomidae)]. AB - Ectinorus (E.) uspallatae n. sp. and Delostichus ojedai n. sp. are described from the rodent Abrocomid, Abrocoma uspallata Braun and Mares, 2002, a new small mammal of Argentina. PMID- 15828581 TI - Entomological and parasitological study on phlebotomine sandflies in central and northern Albania. AB - An entomological survey was carried out in two districts of central (Kruje) and northern (Lezhe) Albania. Six collecting sites, showing a variety of diurnal resting sites, were monitored for adult sandflies from June through October 2002. Flies were collected with CDC miniature light traps, sticky traps and mechanical or hand aspirators in peridomestic sites, in bedrooms and inside cow barns, chicken coops and pigpens. All collecting sites monitored were found positive for sandflies. A total of 849 specimens were caught (29.2% males) belonging to five Phlebotomus species. Phlebotomus neglectus (75.6%) was the most abundant species followed by P. perfiliewi (14.4%), P. papatasi (4.6%), P. tobbi (3.6%) and P. similis (1.8%). The first adult of P. neglectus appeared on June 11 and the last one was collected on October 16. The highest density for this species was observed at the end of July. A total of 111 blood-fed females were caught from the two areas studied. P. neglectus was the only species found blood fed in Lezhe and the same species was prevalent (56.1%) in Kruje followed by P. perfiliewi (30.3%), P. tobbi (10.6%); P. papatasi was represented by only two specimens. Blood meal origin was determined in 45/66 (68.2%) of the females tested from Kruie district. P. neglectus was found fed on four hosts, showing the following feeding patterns: cow (71.4%), dog (117.1%), chicken (5.7%) and human (5.7%); P. perfiliewi was found fed on cow (80.0%) and chicken (20.0%), P. tobbi on cow (50.0%), chicken (25.0%) and dog (25.0%). One specimen of P. papatasi was found fed on cow. When such prevalences were analysed by the available biomass for each host present at the collecting site, P. neglectus resulted to be an opportunistic feeder rather than exhibiting preferences for any specific animal. PCR analysis of 39 P. neglectus from the Lezhe district gave negative results for the presence of Leishmania DNA. PMID- 15828582 TI - [Phlebotomine sandflies from Madagascar (Diptera: Psychodidae). IV--Description of Sergentomyia (Rondanomyia) goodmani n. sp. Reinstatement of the subgenus Rondanomyia Theodor]. AB - This work is the fourth related to species recently described from Madagascar. The authors describe Sergentomyia goodmani on females and males. The female has tubular spermathecae with internal spines. It has an aspect, in Canada balsam, of transversally striped spermatheca, without being completely segmented. The spermathecal ducts share a proximal common part. The pharynx is narrow. The cibarial armature is comb-like. Based on these characters, the authors consider that S. goodmani belongs to the subgenus Rondanomyia. They reinstate this subgenus and consider Neophlebotomus sensu Lewis, 1977 as invalid. Considering Grassomyia as a genus, this is the first record of Sergentomyia in Madagascar. PMID- 15828583 TI - Spread of Leishmania killicki to Central and South-West Tunisia. AB - Twenty cutaneous leishmaniasis (CL) cases were notified from December 2001 to February 2002, in a small village in the district of Oueslatia (governorate of Kairouan, central Tunisia) which is an endemic focus of infantile visceral leishmaniasis due to leishmania (L.) infantum and that had never been concerned previously by CL. The parasite typing of two isolates obtained from two children that have never left the region has identified L. killicki. This species had only been reported previously in a limited focus of Tunisian Southeast. In October 2002, an epidemiological survey with isoenzym characterization of the parasite led in a well-known focus of zoonotic cutaneous leishmaniasis of South-West Tunisia also revealed the presence of L. killicki. These results suggest the spread of this species and stress the need of further investigations for a better control of CL in Tunisia. PMID- 15828584 TI - High infection rates of Triatoma dimidiata are associated with low levels of Trypanosoma cruzi seroprevalence in Pedro Carbo, Ecuador. Use of a tc24 gene based PCR approach. AB - In control programs for vectorial transmission of Chagas' disease, conventional microscopic procedures are generally performed to determine baseline levels of infectivity of vectors. Reported here are data using Polymerase Chain Reaction in the detection of Trypanosoma cruzi in Triatoma dimidiata, one of the principal vectors of Chagas' disease in Ecuador. The microscopy and PCR techniques showed a high percentage of vector infection in Pedro Carbo, province of Guayas (Ecuador), with 44.16% and 46.13% positive insects, respectively. This contrasted with the very low Chagas seropositivity recorded (0.5%). Since T. dimidiata was the only vector of the Chagas' disease found in Pedro Carbo and looking at the vector behavior, our data suggest that despite the high T. dimidiata infection, the low Chagas seropositivity detected is closely associated with the epidemiological and ecological context of T. dimidiata in Pedro Carbo. PMID- 15828585 TI - The phlebotomine sandflies of Portugal. XIII--Occurrence of Phlebotomus sergenti Parrot, 1917 in the Arrabida leishmaniasis focus. AB - In a survey carried out during the summer in 2002 and 2003, in the canine and vulpine leishmaniasis focus of Arrabida, 665 phlebotomine sandflies were caught. 13.83% were P. ariasi, 58.65% P. pemiciosus, 0.45% P. sergenti and 27.07% S. minuta. Despite the low abundance, the finding of the three adults P. sergenti (two males in Aldeia Grande and one female in Quinta da Ramada) confirm the colonization of the Arrabida leishmaniasis focus by this species, which presence had been previously reported but thought accidental. The abundance of P. ariasi observed at this time is significantly different from that in previous years (Pires, 1984 and Fernandes pers. com., 1994). The occurrence of P. sergenti in this region, in association with the decrease in abundance of P. ariasi, may reflect an increased aridness of this region, a consequence of current climate and environmental changes. PMID- 15828586 TI - Efficacy comparison between anti-malarial drugs in Africans presenting with mild malaria in the Central Republic of Africa: a preliminary study. AB - Drug resistance to Plasmodium falciparum contributes to major health problems in central Africa and, as a consequence, poverty. We have analyzed the efficacy of three currently available antimalarial drugs to treat symptomatic, uncomplicated P. falciparum malaria in semi-immune adults living in Bangui, Central Republic of Africa. 210 consecutive individuals were enrolled in the survey, of which 45 were excluded. Those having received dihydroartemisin proved significantly less parasitemic than those having received quinine per os or sulfadoxin-pyrimethamin (chi2 = 16.93; p < 0.05), and 75% recovered in two days compared to 57 and 44%, respectively. The 25% who did not recover benefited from a second cure with dihydroartemisin, which proved 100% efficient. The most accurate protocol remains to be established by analyzing clinical and parasitological data and taking into account the economics of the country. PMID- 15828587 TI - Effect of piroxicam, metamizol, and S-adenosylmethionine in a murine model of experimental trichomoniasis. AB - Biological effects of piroxicam, metamizol, and S-adenosylmethionine (S-AMET) have been tested in NMRI mice infected intraperitoneally with Trichomonas vaginalis. An intraperitoneal treatment during ten preinfection days with piroxicam (10 mg/Kg/day), or metamizol (275 mg/Kg/day), but not with S-AMET (117 mg/Kg/day) induced a significant decrease of abdominal lesions and mortality, assessed by means of a pathogenicity index. The trichomonicidal activity of piroxicam, metamizol, and S-AMET was tested in vitro at the concentration of 300 microM, but found ineffective. These assays have shown the usefulness of the experimental trichomoniasis model for the study of the immunomodulating activity of synthetic drugs. PMID- 15828588 TI - Trypanosoma avium incidence, pathogenicity and response to melarsomine in falcons from Kuwait. AB - Epidemiological and clinical studies on Trypanosoma avium are lacking in the Middle East. The aims of this study were to determine the T. avium incidence in falcons from Kuwait, report clinical signs and find an effective therapy. Blood smears from 921 diseased and 56 healthy falcons were examined between May 2003 and April 2004. 12 birds 11.3%) were found infected by T. avium and ten of these were treated with melarsomine (Cymelarsan) at a dosage of 0.25 mg/kg intramuscularly for four days. All affected birds presented clinical signs, including incapacity of flying high, poor appetite, lethargy, loosing weight, weakness, dyspnoea and death. Signs disappeared within 1-7 days after administration of melarsomine. Trypomastigotes were not detected in blood smears made 1-7 days after the end of therapy. This study suggests that T. avium induces disease in falcons and that melarsomine can be an effective therapy eliminating both clinical signs and circulating trypomastigotes. PMID- 15828589 TI - Labor supply and consumption of food in a closed economy under a range of fixed- and random-ratio schedules: tests of unit price. AB - The behavioral economic concept of unit price predicts that consumption and response output (labor supply) are determined by the unit price at which a good is available regardless of the value of the cost and benefit components of the unit price ratio. Experiment 1 assessed 4 pigeons' consumption and response output at a range of unit prices. In one condition, food was available according to a range of fixed-ratio schedules, whereas in the other condition, food was available according to a range of random-ratio schedules. Consistent with unit price predictions, consumption and response output were approximately equivalent across schedule types within the lower range of unit prices. However, at Unit Prices 64 (ratio value = 192) and greater, considerably more consumption and response output were observed in the random-ratio condition. Experiment 2 replicated these findings with 4 pigeons using the rapid demand curve assay procedure that is commonly used in the behavioral economics literature. Findings are integrated with two mathematical models of behavior under variable reinforcer delays. PMID- 15828590 TI - On the effects of signaling reinforcer probability and magnitude in delayed matching to sample. AB - Two experiments examined whether postsample signals of reinforcer probability or magnitude affected the accuracy of delayed matching to sample in pigeons. On each trial, red or green choice responses that matched red or green stimuli seen shortly before a variable retention interval were reinforced with wheat access. In Experiment 1, the reinforcer probability was either 0.2 or 1.0 for both red and green responses. Reinforcer probability was signaled by line or cross symbols that appeared after the sample had been presented. In Experiment 2, all correct responses were reinforced, and the signaled reinforcer durations were 1.0 s and 4.5 s. Matching was more accurate when larger or more probable reinforcers were signaled, independently of retention interval duration. Because signals were presented postsample, the effects were not the result of differential attention to the sample. PMID- 15828591 TI - Pigeons' choices between fixed-interval and random-interval schedules: utility of variability? AB - Pigeons' choosing between fixed-interval and random-interval schedules of reinforcement was investigated in three experiments using a discrete-trial procedure. In all three experiments, the random-interval schedule was generated by sampling a probability distribution at an interval (and in multiples of the interval) equal to that of the fixed-interval schedule. Thus the programmed delays to reinforcement on the random alternative were never shorter and were often longer than the fixed interval. Despite this feature, the fixed schedule was not strongly preferred. Increases in the probability used to generate the random interval resulted in decreased preferences for the fixed schedule. In addition, the number of consecutive choices on the preferred alternative varied directly with preference, whereas the consecutive number of choices on the nonpreferred alternative was fairly constant. The probability of choosing the random alternative was unaffected by the immediately prior interval encountered on that schedule, even when it was very long relative to the average value. The results loosely support conceptions of a "preference for variability" from foraging theory and the "utility of behavioral variability" from human decision making literatures. PMID- 15828592 TI - Variation, repetition, and choice. AB - Experiment 1 investigated the controlling properties of variability contingencies on choice between repeated and variable responding. Pigeons were exposed to concurrent-chains schedules with two alternatives. In the REPEAT alternative, reinforcers in the terminal link depended on a single sequence of four responses. In the VARY alternative, a response sequence in the terminal link was reinforced only if it differed from the n previous sequences (lag criterion). The REPEAT contingency generated low, constant levels of sequence variation whereas the VARY contingency produced levels of sequence variation that increased with the lag criterion. Preference for the REPEAT alternative tended to increase directly with the degree of variation required for reinforcement. Experiment 2 examined the potential confounding effects in Experiment 1 of immediacy of reinforcement by yoking the interreinforcer intervals in the REPEAT alternative to those in the VARY alternative. Again, preference for REPEAT was a function of the lag criterion. Choice between varying and repeating behavior is discussed with respect to obtained behavioral variability, probability of reinforcement, delay of reinforcement, and switching within a sequence. PMID- 15828594 TI - Emergent, untrained stimulus relations in many-to-one matching-to-sample discriminations in rats. AB - The present experiment investigated whether rats formed emergent, untrained stimulus relations in many-to-one matching-to-sample discriminations. In Phase 1, rats were trained to match two samples (triangle and horizontal stripes) to a common comparison (horizontal stripes) and two additional samples (circle or vertical stripes) to another comparison (vertical stripes). Then, in Phase 2, the rats were trained to match the one sample (triangle) to a new comparison (black) and the other sample (circle) to another comparison (white). In the Phase 3 test, half the rats (consistent group) were given two new tasks in which the sample correct comparison relation was consistent with any emergent stimulus relations that previously may have been learned. The remaining 6 rats (inconsistent group) were given two new tasks in which the sample-correct comparison relation was not consistent with any previously learned emergent stimulus relations. Rats in the consistent group showed more accurate performance at the start of Phase 3, and faster learning to criterion in this phase, as compared with rats in the inconsistent group. This finding suggests that rats may form emergent, untrained stimulus relations between the discriminative stimuli in many-to-one matching-to sample discriminations. PMID- 15828593 TI - Does sensitivity to magnitude depend on the temporal distribution of reinforcement? AB - Our research addressed the question of whether sensitivity to relative reinforcer magnitude in concurrent chains depends on the distribution of reinforcer delays when the terminal-link schedules are equal. In Experiment 1, 12 pigeons responded in a two-component procedure. In both components, the initial links were concurrent variable-interval 40-s variable-interval 40-s, and the terminal links were both 20-s interval schedules in which responses were reinforced by either 4 s of grain in one, or 2-s of grain in the other. The only difference between the components was whether the terminal-link schedules were fixed interval or variable intervals. For all subjects, the relative rate of responding in the initial links for the terminal link that produced the 4-s reinforcer was greater when the terminal links were fixed-interval schedules than when they were variable-interval schedules. This result is contrary to the prediction of Grace's (1994) contextual choice model, but is consistent with both Mazur's (2001) hyperbolic value-added model and Killeen's (1985) incentive theory. In Experiment 2, 4 pigeons responded in a concurrent-chains procedure in which 4-s or 2-s reinforcers were provided independently of responding according to equal fixed time or mixed-time schedules. Preference for the 4-s reinforcer increased as the variability of the intervals comprising the mixed-time schedules was decreased. Generalized-matching sensitivity of initial-link response allocation to relative reinforcer magnitude was proportional to the geometric mean of the terminal-link delays. PMID- 15828596 TI - Using failure mode effects and criticality analysis for high-risk processes at three community hospitals. AB - BACKGROUND: An applied research firm collaborated with staff at three community hospitals to apply Failure Mode Effects and Criticality Analysis (FMECA) to reduce the occurrence of adverse events associated with high-risk processes. The collaboration team, which developed its own FMECA approach, performed FMECAs for six processes, including prevention of patient falls, correct medication ordering and delivery of solid oral medication, and correct blood type transfusion for adult medical surgery patients. DEVELOPMENT OF FMECA PROCEDURE AND TOOL: The hospitals followed eight specific steps to gather data, conduct FMECA sessions, and identify medical process weaknesses and risk reduction measures. Worksheets, including each step of the system process, success criteria, possible failure modes, causes of failure, frequency of failure, consequence of failure, and safeguards placed to avoid failure, were used to capture information during the FMECA sessions. CONCLUSIONS: On the basis of identified weaknesses, along with cost and other administrative considerations, medical process improvements were devised. Rules for devising improvements included improvements that help prevent the failure mode were better than those that mitigate the consequences, passive features that prevent failures were better than administrative controls, and improvements with the highest reliability were favored. PMID- 15828595 TI - Learning to improve safety: false-positive pathology report results in wrongful surgery. AB - BACKGROUND: A patient experienced a wrongful surgical resection, specifically, a radical retropubic prostatectomy because of a false-positive pathology report. FINDINGS FROM THE ROOT CAUSE ANALYSIS (RCA): The RCA team identified three antecedent events that contributed to this medical error: (1) a second (concurring) pathologist did not provide a written opinion, (2) a single pathologist who reviewed and signed the final report, and (3) a pathologist who did not review the case and reconfirm the diagnosis immediately prior to the surgical resection. RECOMMENDATIONS: The RCA team recommended that the concurring pathologist write his or her diagnostic findings on the referral form, two pathologists review and sign the final typed report, and a pathologist rereview the slides on the business day prior to a surgical resection. Because the prostate specific antigen (PSA) value can be helpful in select cases of prostate cancer, the team recommended the PSA value be referenced when reviewing prostate specimens obtained through fine-needle biopsy. TRACKING COMPLIANCE: Because a wrongful surgical resection is a rare event, emphasis was placed on measuring compliance with distinct elements that were part of the revised procedure. During a 12-month span, practitioners demonstrated sustained compliance to the enhanced process for analyzing and reporting results. PMID- 15828597 TI - New insulin infusion protocol Improves blood glucose control in hospitalized patients without increasing hypoglycemia. AB - BACKGROUND: Aggressive treatment of hyperglycemia in hospitalized patients can improve clinically important outcomes. At the University of Washington Medical Center a quality improvement project was conducted to develop and implement a standardized insulin infusion protocol for use throughout the institution. METHODS: The insulin infusion protocol was piloted on critical and non-critical care inpatient units. Safety and efficacy data were collected for a one-month period on each unit. RESULTS: A total of 156 patients were evaluated. The incidence of hypoglycemia was lower for the study group than the historical controls, as was the mean percentage of time patients were hyperglycemic, 15% +/- 2.3% vs. 33% +/- 2.6% for the critical care subgroup (p < .0001) and 18% +/- 2.4% vs. 56% +/- 2.3% for the non-critical care subgroup (p < .0001). CONCLUSIONS: The insulin infusion protocol better met the insulin requirements of our patients and achieved better glycemic control than previous protocols at the institution. In addition, there was no increase in hypoglycemia despite the use of the protocol in non-critical care units with higher patient-to-nurse ratios, suggests that insulin infusion therapy can be safely used outside of critical care units. PMID- 15828598 TI - Unintended consequences of a quality improvement program designed to improve treatment of alcohol withdrawal in hospitalized patients. AB - BACKGROUND: New guidelines, accompanied by an educational campaign, introduced standardized monitoring of withdrawal severity while emphasizing prophylactic fixed-schedule benzodiazepine (BDZ) treatment of at-risk patients. EVALUATION: Preliminary analysis showed more deaths during the year after introduction of the guidelines. Investigation revealed some evidence of guideline adherence and a decrease in the number of patients requiring transfer to a higher level of care. However, an 18% increase in the median length of stay was also found, as was an increase in the total dose of benzodiazepines administered to patients with cirrhosis and severe concurrent illness, and the risk of in-hospital death persisted even after adjustment for patient mix. RESPONSE: This feedback led to guideline revision and redoubled educational efforts focused on safe benzodiazepine prescribing. Ongoing monitoring of patient outcomes showed no further deterioration and some evidence of improved quality of care. CONCLUSION: Evaluation of such quality improvement efforts should include measurement of both treatment patterns and patient outcomes. PMID- 15828599 TI - Do on-site patient satisfaction surveys bias results? AB - BACKGROUND: Response rates, patient sample characteristics, and patient satisfaction ratings were compared between two surveying methods: (1) surveys completed at the physician office site (on-site surveying), and (2) surveys mailed to patient homes following the encounter (mail-out/mail-back). METHODS: Surveying was completed at three physician practices within a 214-physician medical practice. Patients with physician appointments during four-hour time blocks were randomly split to receive either on-site or mail-based satisfaction surveys. RESULTS: Participants younger than 45 years of age provided much higher satisfaction ratings on site than they did by mail (p < .0001), and participants older than 45 years of age reported satisfaction levels consistently whether on site or by mail. Both age groups reported higher satisfaction with "people aspects" of care on site than they did by mail (p < .001). DISCUSSION: On-site methods may yield satisfaction results that are biased in a positive direction for younger patients and for all patients in which social desirability pressures are prominent. Therefore, organizations that rely on such information may have an inflated view of the patient's satisfaction with their care delivery experience. Secondly, because the differences in ratings are the greatest for the "people aspects" of care, if improvement efforts are prioritized on the basis of these rapid results, the wrong priorities may be set. PMID- 15828600 TI - A systems approach to identifying inappropriate use of controlled substances: the need for balance. AB - BACKGROUND: The diversion, abuse, and inappropriate use of controlled substances remain of concern among health care practitioners, insurers, and policy makers in the United States. The principle of "balance" represents a need to establish a system of control to prevent abuse, trafficking, and diversion of controlled substances, while ensuring their medical availability for legitimate patients. ADMINISTRATIVE DATABASES TO STUDY OUTCOMES AND QUALITY OF CARE: Some models employ pattern recognition within administrative health databases, for example, to examine outcomes and quality of medical care and monitor fraudulent behaviors and inappropriate medical care. Patients' use of resources or variations in practitioners' practice patterns can be examined. A SYSTEMS APPROACH TO ADDRESS INAPPROPRIATE USE OF CONTROLLED SUBSTANCES: A systems approach would require collaboration with medical and pharmacy directors, systems analysts, coding experts, legal experts, and clinicians to develop the claims-based model. Once a patient and/or practitioner with possible diversion or abuse are identified, a second step is required to distinguish inappropriate and appropriate behaviors and medical care. CONCLUSION: Programs to detect misuse of controlled substances must be validated through clinical research, and a consensus should be reached as to what constitutes a breach of accepted medical practice. PMID- 15828601 TI - Reducing the risk of health care-associated infections by complying with CDC hand hygiene guidelines. AB - BACKGROUND: The Joint Commission on Accreditation of Healthcare Organizations has made the reduction of health care-associated infections one of its National Patient Safety Goals for 2005. CASE STUDY: A 57-year-old man who underwent coronary artery bypass graft surgery was discharged from the hospital without any complications. During his routine follow-up surgery clinic visit two weeks later, his sternal wound was found to be infected. Wound and blood cultures were positive for methicillin-resistant Staphylococcus aureus (MRSA). IMPLEMENTING SOLUTIONS: Health care workers at Barnes-Jewish Hospital (BJH), St. Louis, attended a mandatory educational session that reviewed hand hygiene guidelines and the hospital policy related to hand hygiene and infection control. In addition, posters and flyers were posted. An increase in the use of alcohol hand rubs was found. In a later effort, a multidisciplinary hand hygiene campaign was launched at one of the BJC community hospitals. All physicians were sent hand hygiene materials and supplies, and signs were posted in key areas such as the hospital cafeteria. After a six-month period, the compliance rate increased from 35% to 53%. CONCLUSION: Hospital-related infections will never be completely eradicated, but many can be prevented. PMID- 15828602 TI - Periodontal plastic surgical indications in the restorative practice. AB - A variety of advanced surgical techniques are available today for the treatment of soft-tissue complications in the restorative patient. Gingival recession and lack of attached gingiva can complicate delicate esthetic and prosthetic treatment. The restorative office must be able to critically evaluate the demands of the case and offer the patient options for correction of these advanced conditions. The ability to counsel in an informed way-outside the restorative arena-allows dentists to demonstrate their true commitment to the best outcome for the person they are dedicated to serve, the patient. PMID- 15828603 TI - Peer review is the way to go. PMID- 15828604 TI - The art and science of tooth whitening. AB - Although tooth whitening is one of the most popular dental procedures, it is also one of the least understood. We are still unclear as to its mechanism of action. There is little data as to the effects of both concentration and dose on outcome. The techniques for measuring color change have been brought into question. The cause(s) of sensitivity and the effects of long-term exposure to hydrogen peroxide are not clear. The issue of rebound in color has not been well examined, and issues related to maintenance of the whitening effect are also poorly understood. The immediate placement of composite resin on bleached teeth has been controversial. According to a study evaluating the shear bond strength of composite restorations placed on bleached and nonbleached teeth, there was no statistically significant difference when the composites were placed at 24 hours, 48 hours, four days, or six days. A different study, which evaluated the effects of take-home bleaching systems on enamel surfaces, suggests that a period of four days must elapse before bonding to a tooth bleached with a peroxide material, while no delay is necessary for a non-peroxide-based bleaching system. Most recently, there has been a push to find ways to accelerate and improve the delivery of the whitening process. These include the application of a number of different light sources believed to accelerate the breakdown of peroxide and thus speed up the whitening process. However, the research in this area has been controversial, with publications having quite different conclusions as to the efficacy of light-activated bleaching. Finally, the issues of adverse events and possible side effects were reviewed. The toxicological side effects of tooth bleaching systems seem to be minimal. However, tooth sensitivity can be quite significant. Although its causes are poorly understood, tooth sensitivity is most often seen as the result of tooth dehydration. So while patient demand for tooth whitening is at an all-time high, and dentists have more options for treatment, it's important that dentists evaluate which of these options is most ideal for their patients, factoring in the patients' cost and time issues, as well as sensitivity to the procedure. PMID- 15828605 TI - Assisting the dentist with chemical dependency: an overview. PMID- 15828606 TI - Applications of digital images derived from toothprints thermoplastic bite impression wafers. PMID- 15828608 TI - Much ado about digital radiography. PMID- 15828607 TI - A clinico-pathologic correlation. PMID- 15828609 TI - Recurrent intraoral herpes infection. PMID- 15828610 TI - Sweet teeth. PMID- 15828611 TI - [Auto-inflammatory syndromes]. AB - Auto-inflammatory syndromes are a group of hereditary diseases characterised by intermittent bouts of clinical inflammation with focal organ involvement mainly: abdomen, musculoskeletal system and skin. The most frequent is familial Mediterranean fever, which affects patients of Mediterranean descent all over the world. Three other types have been recently clinically as well as genetically characterised. A thorough diagnosis is warranted, as clinical and therapeutic management is specific for each of these diseases, as underlied by a specific inflammatory pathway. This new group of diseases has already opened new avenues in our understanding of the inflammatory response. PMID- 15828612 TI - [Menopause: a different challenge for each woman]. PMID- 15828613 TI - [What is menopause?]. AB - Menopause is a natural phenomenon consisting in follicles atresia and decrease in ovarian hormonal secretions. The mean age of menopause is 51 years. It is known that follicles atresia is partially explained by genetics factors. During this period, we observe hormonal variations. The most important of them are an elevation of FSH level, a decrease in estrogens. We also note a variation of androgens secretion. Hypo-oestrogenia induces various clinical symptoms. Some of them occur early in menopause (climateric syndrom), others occur later (osteoporosis, cardiovascular diseases). PMID- 15828614 TI - [Historical perspective of hormone replacement therapy]. AB - Clinical manifestations of menopause are partly related to estrogen deficiency. Estrogen replacement was long believed to reverse not only climacteric symptoms but also other chronic conditions associated with menopause such as osteoporosis, cognitive disorders or cardiovascular risk. However, it rapidly became obvious that hormone replacement therapy (HRT) was also associated with an increased risk of breast cancer and venous thrombo-embolic events. Until the end of the 1990's, based on cohort studies, HRT was thought to prevent cardiovascular complications of atherosclerosis. Risk/benefit ratio was thus considered as in favor of HRT, explaining its very wide prescription, after specific contra-indications have been ruled out. In 1998, the publication of HERS, the first randomized controlled study evaluating the effects of HRT in secondary cardiovascular prevention, allowed the scientific community to be conscious of the fact that HRT, not only did not prevent cardiovascular risk but also, probably, aggravated it. In 2002, the premature interruption of WHI study, by confirming that this was also true for primary prevention, has profoundly altered the common beliefs about HRT. Indeed, if HRT was associated with an increased cardiovascular risk, the benefits/risks ratio became unfavorable. Since that time, less women are treated and some of them have stopped their HRT. Recent recommendations have been published about indications of HRT, mainly based on the presence of climacteric symptoms. The potential interest of transdermic route for administration of estrogens needs to be confirmed. The potential deleterious effect of progestins needs to be explored. The difficult story of HRT had, at least, the merit to show, one more time, that in medicine, scientific evidence is always better than beliefs. PMID- 15828615 TI - [Hormonal replacement therapy and breast cancer]. AB - The recent publication of some randomised control trials results and of two important cohort studies makes possible a better understanding of the relation between hormonal replacement therapy (HRT) and breast cancer. It is now well established that estrogen-progestin replacement therapy is associated with an elevation of breast cancer risk. It is not possible to define duration of treatment without risk. Data for estrogens alone are discordant. Five years after stopping HRT, the risk will join the level of non-users. By now, HRT may be prescribed in women that suffer intense climacteric symptoms, and after careful information about benefits, risks, and mammary effects of HRT. The minimal dose of estrogens is required for the control of menopausal symptoms. Estrogens alone will be prescribed in hysterectomised women. Mammography has to be performed before the beginning of HRT, and then every two years, in the absence of important risks factors of breast cancer. The indication of HRT has to be discussed every year, and some therapeutic pauses must be planned. In case of increased breast density at mammography, ultra-sonography may be useful. Breast surveillance must go on after HRT termination. PMID- 15828616 TI - [Hormonal treatment for menopause and arterial risk]. AB - Is hormonal atmosphere before menopause the cause of the lower risk of coronary heart disease in women? Big clinical trials do not validate this hypothesis, however simple and attractive: in 5 primary or secondary prevention trials, estrogens alone or in association with progestatives to near 32000 women after menopause, do not leave hope for a significant reduction of coronary risk, and show in contrast an 30% increase of stroke risk. These results highlight the crucial importance of clinical trials to validate therapeutic models. The remaining hypotheses on the nature of hormonal treatments and the administration route must follow the same validation process. The prescription of hormonal treatment for menopause illustrates the importance of informed decision including individualised estimate of the risk to benefit ratio. PMID- 15828617 TI - [Hormone therapy and risk for venous thromboembolism in postmenopausal women]. AB - Oral estrogens are associated with a two-fold to three-fold increased risk of venous thromboembolism in postmenopausal women. Absolute excess risk is low but higher than for contraceptive pills. Biological data and observational studies suggest that transdermal estrogens are associated with a lower risk for venous thromboembolism than oral estrogens but safety has yet to be proved. Whether addition of progestin plays a significant role is undetermined so far. PMID- 15828618 TI - [Management of postmenopausal osteoporosis]. AB - Current management of postmenopausal osteoporosis has benefited from several advances both in the screening or those women with the highest risk of fracture and the development of efficient drugs to reduce the occurrence of fracture. At the individual level, assessment of the risk of fracture must associate the level of bone mineral density which represents the major determinant of fracture and several clinical risk factors. Measurements of biochemical markers of bone turnover are sometimes useful to improving this risk evaluation. However, the 5- to 10-year absolute risk or fracture still needs to be determined as well as the threshold above which a therapeutical intervention should be warranted. Besides hormone replacement therapy, which indications are now more limited than before, the different therapeutical options include several anti-osteoclastic drugs and more recently new anabolic compounds. It should be now possible to anticipate a long term strategy for the prevention of osteoporosis based on an early screening of the women at higher risk of fracture as well as the optimal choice of treatment taking into account the age and underlying clinical conditions of each woman. PMID- 15828619 TI - [Monitoring of postmenopausal women]. AB - Postmenopausal age women have an increased risk of cardiovascular diseases, osteoporosis, and gynaecological cancers. A regular follow up is necessary to evaluate the interest of HRT, and for screening cardiovascular diseases and gynaecologic cancers. Patient counselling includes life style modifications, follow-up and regular evaluation of risk factors and potential benefit of HRT for each woman. PMID- 15828620 TI - [Menopause: what do you feel?]. PMID- 15828621 TI - [EBM in practice]. PMID- 15828622 TI - [Tumours of the buccal cavity and the upper respiratory-digestive tracts]. PMID- 15828623 TI - [Arterial hypertension in the adult]. PMID- 15828624 TI - [Shock]. PMID- 15828625 TI - [Personality disorder]. PMID- 15828626 TI - [Problems of intra-cardiac conduction]. PMID- 15828627 TI - [Ludwik Rajchman (1881-1965), Polish doctor and citizen of the world]. PMID- 15828628 TI - MedPAC urges Congress to move on pay for performance. PMID- 15828629 TI - Inspiratory resistance as a potential treatment for orthostatic intolerance and hemorrhagic shock. AB - Loss of consciousness due to central hypovolemia can occur due to sudden cardiovascular decompensation in normal individuals or hypovolemic shock in wounded patients. A variety of devices have been developed to sustain perfusion to the brain including anti-G suits worn by pilots and returning astronauts and applied to patients as "shock trousers." However, all countermeasures developed to date suffer from problems that limit their utility in the field. An "impedance threshold device" (ITD) has recently been developed that acutely increases central blood volume by forcing the thoracic muscles to develop increased negative pressure, thus drawing venous blood from extrathoracic cavities into the heart and lungs. We review here a series of experiments that demonstrate the application of the ITD to a variety of experimental conditions, including its use to: 1) increase heart rate, stroke volume, and arterial BP in normovolemia and hypovolemia; 2) increase cerebral blood flow velocity; 3) reset cardiac baroreflex function to a higher operating range for BP; 4) lower intracranial pressure; and 5) reduce orthostatic symptoms. In this brief review, we present evidence which supports further consideration of using inspiratory resistance as a countermeasure against circulatory collapse associated with orthostatic instability and hemorrhagic shock. PMID- 15828630 TI - Exercise ending 30 min pre-dive has no effect on bubble formation in the rat. AB - INTRODUCTION: We have previously shown that exercise performed 20 h before a dive significantly reduces bubble formation in both rats and humans. Furthermore, exercise performed closer to the dive did not prevent bubble formation. HYPOTHESIS: The present study was designed to determine whether exercise 30 min prior to a dive promotes bubble formation. The occurrence of many bubbles is linked to a higher risk of developing decompression sickness. METHODS: A total of 58 Sprague-Dawley rats were randomly divided into a sedentary control group (n = 29) and an exercise group (n = 29). Rats in the exercise group ran on a treadmill for a total of 90 min at variable intensity up to 85-90% of VO2max. Then, 30 min after exercise, one rat from each group rested in a pressure chamber at 700 kPa (7 atm) breathing air, performing a simulated dive. Bottom time was 45 min; decompression rate was 50 kPa x min(-1) (0.5 atm x min(-1)). Immediately after surfacing (100 kPa, 1 atm), the rats were anesthetized and bubbles were measured discontinuously for the next 60 min. RESULTS: There were no significant differences in survival (p = 0.55), median bubble grade (p = 0.67), survival time (p = 0.53), or the number of rats getting a bubble score > or = 2 (p = 0.79) between the groups. CONCLUSION: The same type and intensity of exercise that reduces bubble formation when performed 20 h prior to a dive neither promotes nor reduces bubble formation if performed 30 min before a dive. The present data indicate that exercise completed 30 min before a dive does not increase the risk of developing decompression sickness in the rat. PMID- 15828631 TI - Modeling steady-state intracranial pressures in supine, head-down tilt and microgravity conditions. AB - INTRODUCTION: The effects of microgravity are often simulated by head-down tilt (HDT). While data exist for intracranial pressure (ICP) during short-term HDT, no corresponding data exist for long-term exposure to HDT or microgravity. A mathematical model was developed to predict these unknown long-term responses. Predicted pressures include those in the cerebral vasculature, ventricular and extra-ventricular cerebrospinal fluid (CSF), and the brain tissue extracellular fluid. METHODS: A mathematical model was used to predict steady-state responses to various stimuli. Simulated CSF infusion tests were used to estimate model parameters such as the filtration coefficient of the cerebral capillary bed. Short-term HDT simulations validated the model. Further simulations predicted ICP responses to long-term HDT and microgravity. RESULTS: Constant-rate infusion simulations predict that the filtration coefficient of the cerebral capillary bed is between 4.46 and 5.15 x 10(-3) {[(ml x min(-1)) x mmHg(-1)] x 100 g(-1)}. Short-term HDT simulations reproduced clinical observations for venous sinus pressure and ICP. Further simulations produced the following relationships: first, ICP is unaffected by the expected changes in central artery pressure. Second, ICP changes in parallel with central venous pressure. Third, ICP increases 0.37 mmHg per 1 mmHg decrease in blood colloid osmotic pressure. DISCUSSION: Results suggest that despite the presence of tight capillary junctions in the brain, the cerebral capillary filtration coefficient is of the same order of magnitude as measured in the calf and forearm. Simulations also suggest that ICP in microgravity is significantly less than that in long-term HDT and may be less than that in the supine position on Earth. PMID- 15828632 TI - Isometric force production in high Gz: mechanical effects, proprioception, and central motor commands. AB - BACKGROUND: We have shown in the past that human subjects produce exaggerated isometric responses when exposed to high +Gz. The present study investigated the role played by different factors in this phenomenon. METHODS: There were 12 healthy non-pilot volunteers who were seated in the gondola of a centrifuge and were exposed to a +1, +1.5, and +3 Gz environment. During each exposure, they produced forces of prescribed magnitudes and directions in their frontal plane using an isometric joystick. From the registered data, we determined initial (100 ms after onset), peak, and end (70 ms before trial end) force. RESULTS: For a given target magnitude, initial, peak, and end force were each smallest in 1 G, higher in 1.5 G, and higher still in 3 G. This G-related exaggeration was most pronounced for initial, and less for peak and end force. We further found that responses in high G were biased downwards for end force, but not for initial and peak force. CONCLUSIONS: The G-related excesses of produced force are probably due to less appropriate central commands, since it manifests before proprioception becomes effective. The amelioration of this deficit for peak and end force is probably achieved by partial corrections based on proprioceptive feedback, or by direct mechanical effects. Our findings are relevant for the safe operation of high-performance aircraft during high-G maneuvers. PMID- 15828633 TI - EEG and ECG changes during simulator operation reflect mental workload and vigilance. AB - BACKGROUND: Performing mission tasks in a simulator influences many neurophysiological measures. Quantitative assessments of electroencephalography (EEG) and electrocardiography (ECG) have made it possible to develop indicators of mental workload and to estimate relative physiological responses to cognitive requirements. OBJECTIVE: To evaluate the effects of mental workload without actual physical risk, we studied the cortical and cardiovascular changes that occurred during simulated flight. METHODS: There were 12 pilots (8 novices and 4 experts) who simulated a flight composed of 10 sequences that induced several different mental workload levels. EEG was recorded at 12 electrode sites during rest and flight sequences; ECG activity was also recorded. Subjective tests were used to evaluate anxiety and vigilance levels. RESULTS: Theta band activity was lower during the two simulated flight rest sequences than during visual and instrument flight sequences at central, parietal, and occipital sites (p < 0.05). On the other hand, rest sequences resulted in higher beta (at the C4 site; p < 0.05) and gamma (at the central, parietal, and occipital sites; p < 0.05) power than active segments. The mean heart rate (HR) was not significantly different during any simulated flight sequence, but HR was lower for expert subjects than for novices. The subjective tests revealed no significant anxiety and high values for vigilance levels before and during flight. CONCLUSIONS: The different flight sequences performed on the simulator resulted in electrophysiological changes that expressed variations in mental workload. These results corroborate those found during study of real flights, particularly during sequences requiring the heaviest mental workload. PMID- 15828634 TI - Preflight virtual reality training as a countermeasure for space motion sickness and disorientation. AB - INTRODUCTION: Research suggests that preflight training in virtual reality devices can simulate certain aspects of microgravity and may prove to be an effective countermeasure for space motion sickness (SMS) and spatial disorientation (SD). It is hypothesized that exposing subjects preflight to variable virtual orientations, similar to those encountered during spaceflight, will reduce the incidence and/or severity of SMS and SD. METHODS: Subjects were assigned to either a variable training (VT) or nonvariable training (NVT) condition to perform a simple navigation and switch activation task in a virtual space station. VT subjects performed the task starting in several different orientations, whereas NVT subjects always performed the task starting in the same orientation. On a separate day, all subjects then performed the same task in a transfer of training session starting from a novel orientation. RESULTS: When exposed to the novel test orientation, VT subjects performed the tasks more quickly (12%) and with fewer nausea symptoms (53%) than during the training session, compared with NVT subjects who performed more slowly (6%) and with more nausea symptoms (28%). Both VT and NVT conditions were effective in reducing the number of wall hits in the novel orientation (39% and 34%, respectively). DISCUSSION: These results demonstrate the effectiveness of using variable training in a virtual environment for reducing nausea and improving task performance in potentially disorienting surroundings, and suggest that such training may be developed into an effective countermeasure for SMS, SD, and associated performance decrements that occur in spaceflight. PMID- 15828635 TI - Midair collisions: limitations of the see-and-avoid concept in civil aviation. AB - INTRODUCTION: Midair collisions cause loss of life and property and undermine general aviation; they also represent failures of the see-and-avoid concept in air traffic control. This study identifies limitations of the see-and-avoid concept. METHODS: Analysis of National Transportation Safety Board data on 159 U.S. civil aviation midair collisions and limitations of the see-and-avoid concept. RESULTS: On average, there were 15.6 midair collisions annually from 1991 through 2000. At least one aircraft was maneuvering in 88% of collisions, and both in 70%. There were 77% that involved arrival to, departure from, or flight over an airport, with 61% in the traffic pattern. Head-tail collisions were more frequent in the traffic pattern than out (28.3%, 2.8%, p < 0.05). Other horizontal convergence angles were equally frequent in or out of the pattern: head-on (8.3%, 11.0%), obtuse (11.0%, 8.3%), and acute (13.8%, 16.6%). DISCUSSION: Because the relative bearing to each aircraft on an unaccelerated collision course is constant, pilots sometimes cannot see converging aircraft when climbing, descending, or level. Even if a converging aircraft is unobstructed, it appears small, motionless, camouflaged, and inconspicuous until imminent impact. A statistical model reveals that the probabilities of seeing and avoiding a converging 40-ft aircraft, for an optimal observer or theoretical pilot scanning 2/3 or 1/3 of the time, respectively, are less than 0.91, 0.60, 0.30 at 200 kn; 0.49, 0.32, 0.16 at 300 kn; 0.28, 0.18, 0.09 at 400 kn; and 0.15, 0.10, 0.05 at 500 kn. The see-and-avoid concept has striking physical and behavioral limitations. PMID- 15828636 TI - Hearing thresholds among young professional divers: a 6-year longitudinal study. AB - INTRODUCTION: Diving is associated with pressure effects and noise exposure that can impair hearing. We were interested in finding out whether changes in hearing thresholds were related to cumulative exposure in young professional divers. METHODS: In a prospective follow-up design, we studied a cohort of 47 men from the beginning of their education as professional divers at a mean age of 25.6 yr (range 18-38 yr). They had audiograms taken at baseline, after 1 and 3 yr, and finally after 6 yr, during which they performed 40-1458 (median 184) sea dives, most of them to a depth of 10 m or less. The hearing tests measured pure tone air conduction thresholds at 0.25-8.0 kHz in a sound-treated booth. Annual change of hearing thresholds were calculated by fitting linear regression lines to individual data. For purposes of analysis, the divers were divided into two groups, low-exposure (199 or fewer dives, n = 26) and high-exposure (200 or more dives, n = 21). RESULTS: Hearing was better in the divers than in the general population at baseline and follow-up. At 6 yr, significant reductions were found in the divers' hearing at 4 kHz and 8 kHz. Significant increases in thresholds were found at 0.5, 2, 3, and 6 kHz for the right ear and 3, 4, and 6 kHz for the left ear. The annual change at 4 kHz was 0.95 dB in the left ear and 0.18 dB in the right ear (p = 0.01). There was no significant difference in hearing changes between the high- and low-exposure groups. DISCUSSION: The results suggest that a mild hearing impairment can occur in young professional divers due to occupational and other noise exposure. PMID- 15828637 TI - G-induced loss of consciousness: case-control study of 78 G-Locs in the F-15, F 16, and A-10. AB - INTRODUCTION: This study determined the trends of reported G-induced loss of consciousness (G-LOC) mishaps from 1980--1999, and determined potential risk factors in pilot characteristics; specifically, 30/60/ 90-h and sortie history, total flight hours, total hours in the aircraft, age, height, weight, and BMI. METHODS: Using aircraft malfunction mishaps to reflect a cross-section of USAF pilots, potential risk factors were determined using a case-control method; cases were all G-LOC mishaps and controls were aircraft malfunction mishaps. The data consisted of 2002 mishap pilots in the history of the F-16, F-15, F-15E, and A-10 from 1980-1999. RESULTS: During this time, G-LOCs represented only 2.5% of all mishaps. The mean engagement number for G-LOC mishaps was three at an average of 8 Gs. A poor anti-G straining maneuver was cited in 72% of the mishaps, fatigue and G-suit malfunction in 19%, low G-tolerance at 14%, and 37% were student pilots. Within pilot characteristics, only two factors were found to be statistically significant: the time in the aircraft and pilot age. In the F-16, there was a 3.5 times greater chance of experiencing a G-LOC mishap if the pilot had less than 600 h in the aircraft [3.5 (1.7-7.2, 95%CI)], and a 9.5 times greater chance in the F-15 [9.5 (2.2-41.9, 95%CI)]. There was a 4.5 times greater chance of experiencing a G-LOC mishap if under the age of 30 in the F-16 [4.5 (2.3-8.5, 95% CI)] and a 3 times greater chance in the F-15 [2.8 (1.2-6.8, 95% CI)]. DISCUSSION: Though it is difficult to predict who will experience G-LOC, emphasis on prevention must be concentrated in training and in pilots new to the aircraft. PMID- 15828638 TI - Neck strength and myoelectric fatigue in fighter and helicopter pilots with a history of neck pain. AB - INTRODUCTION: Flight-induced neck pain at high Gz loads or during sustained rotary-wing missions may be caused by limitations in neck muscle function. A better understanding of the contributing factors of excessive external load and internal neck-stabilizing mechanisms would improve the ability to prevent and treat such pain. The aim of this single-blinded cross-sectional study was to evaluate neck neuromuscular function in fighter and helicopter pilots who suffered from frequent neck pain. METHODS: Subjects with pain were 16 fighter pilots (FP-P) and 15 helicopter pilots (HP-P) with frequent neck pain episodes who were compared with pain-free controls (FP-C and HP-C). In all groups, neck extensor and flexor muscles were studied by measuring 1) the strength of maximum voluntary contraction (MVC), and 2) fatigue due to a submaximal isometric contraction. The decline (slope) of the electromyogram (EMG) median frequency power spectra was used as an index of fatigue, while initial median frequency (fi) was taken from the intercept of the regression line. RESULTS: Two-way analysis of variance (ANOVA) revealed interaction effects for extensor MVC. Post hoc testing showed that FP-P had significantly lower extensor MVC (p = 0.03) than FP-C, while there was no such difference for the HP-P vs. HP-C or between the two control groups. There were no significant effects for MVC-balance (flexors/extensors); nor were there any fi or extensor EMG-slope effects. However, there were interaction effects for flexor EMG-slopes: HP-P showed lower slopes than did HP-C (p = 0.02). CONCLUSIONS: To protect and stabilize the head and neck in high Gz environments, higher neck muscle strength is needed; less muscle strength in FP-P may cause further pain and perhaps reduced mission effectiveness. Less localized steep slopes for HP-P might reflect impaired muscle functioning. Specific preventive and clinical attention may be warranted for different types of pilot. PMID- 15828639 TI - Mechanical counter-pressure vs. gas-pressurized spacesuit gloves: grip and sensitivity. AB - INTRODUCTION: An elastic mechanical counter pressure (MCP) glove for spacesuits is under development. In this study we compared handgrip and pinch grip strength levels for the MCP glove and the current extravehicular mobility unit (EMU) gas pressurized glove. METHODS: We employed handgrip and pinch grip dynamometers to assess strength levels and von Frey monofilaments to evaluate hand sensitivity. Tests were conducted with the gloved hand inserted in an evacuation chamber at 200 mmHg below atmospheric pressure to simulate conditions in space. RESULTS: Average bare hand strength was 463 N and decreased to 240 N for EMU and 250 N for MCP. Pinch grip and key grip testing showed no difference among conditions. However, there was a significant decrease in palmar grip strength from 111 N barehanded to 67 N in both gloves. Barehanded endurance time was 160 s and dropped to 63 and 69 s for EMU and MCP, respectively. Sensitivity was significantly better for MCP compared with the EMU. DISCUSSION: The MCP glove improved hand sensitivity when compared with the EMU glove and performed as well as the EMU glove in terms of overall handgrip strength, endurance at 25% of maximum handgrip strength, pinch grip, palmar grip, and key grip tests. Improvements in fabric composition and glove design may further improve ergonomic and other functional parameters of the MCP glove. PMID- 15828640 TI - Assessment of simulated surgical skills in parabolic microgravity. AB - BACKGROUND: During spaceflight crew health is paramount in the success of flight missions. The delivery of healthcare during flight requires crew readiness for medical and surgical response. METHODS: There were 20 participants who were evaluated for accurate performance of 4 basic laparoscopic surgical skills (clip applying, cutting, grasping, and suturing) during parabolic weightlessness using an inanimate workstation aboard the NASA KC-135 aircraft. RESULTS: Data indicate that motor skill performance decreased within the parabolic microgravity flight environment. Performance in parabolic microgravity flight included futile effort with an increase in number of tasks attempted and a decrease in tasks completed successfully. CONCLUSIONS: There is a decreased frequency of accurate task completion in parabolic microgravity flight, but it is not an obstacle to implementation of effective training for providing in-flight medical care. The data reveal that individuals perform basic laparoscopic surgical simulation with greater effort in microgravity following simulation training. PMID- 15828641 TI - Altitude training experiences and perspectives: survey of 67 professional pilots. AB - INTRODUCTION: Pilots and crewmembers of flights exceeding 7620 m/mean sea level (msl) are required to complete ground training in high-altitude physiology, including hypoxia training. However, regulations do not require altitude chamber training (ACT). METHOD: An anonymous questionnaire concerning their experiences and perceptions of hypoxia training was filled out by 67 pilots attending an aviation safety conference. All pilots had logged professional business flight hours in the previous 6 mo. RESULTS: There were 62 pilots who reported receiving hypoxia training, and of these, 71% reported having initial ACT. Most of the pilots surveyed agreed that all pilots should receive introductory hypoxia training (92%), recurrent hypoxia training (86%), initial ACT (85%), and recurrent ACT (70%). Initial ACT received lower endorsements for private (32%) or recreational (10%) pilots than for commercial (74%) and air transport (90%) pilots. When asked if ACT should be based on the altitude capability of an aircraft, 59% responded affirmatively. Apparently, the perceived need for ACT was based on the likelihood of flying at higher altitudes and not simply the level of certification. When asked if the current regulations (i.e., not requiring ACT) addressing high-altitude flying (above 7620 m/msl) are sufficient, 52% of the current sample disagreed or strongly disagreed. DISCUSSION: Generally, these professional pilots perceived that pilot training should include introductory hypoxia training, recurrent hypoxia training, and ACT. Exceptions were initial ACT for recreational pilots and private pilots. Generalizability of these results may be affected by the specificity and size of the sample. Distributing the survey to a wider audience of pilots would provide additional information regarding perceptions of hypoxia training. PMID- 15828642 TI - Middle East aircrew use of alcohol, tobacco, coffee, and medicaments. AB - OBJECTIVE: The purpose of this study was to assess the prevalence of the use of alcohol, tobacco, coffee, and various medications among flying crew and clerks working in a ministry in Kuwait. METHODS: In a cross-sectional study, a self administered questionnaire was filled in by 277 flying crew and 302 clerks of a comparable age and gender ratio. RESULTS: Frequencies of using alcohol, tobacco, coffee, and various medications were much higher among flying crew as compared to clerks. Flying crew were a very homogenous group in terms of exposure to the substances studied with only two exceptions: younger people were more likely to drink alcohol abroad (but not in Kuwait), and cabin crew relied on herbal medicine more than pilots. CONCLUSIONS: Flying crew differ very much from a socially comparable segment of the local population, but it is not possible to discern the impact of occupational risks (jet leg and night shifts) from exposure to foreign influences and, as for alcohol and sedatives, from easier availability. PMID- 15828643 TI - Heat stress and carbon monoxide exposure during C-130 vehicle transportation. AB - INTRODUCTION: Running gasoline engines in a confined space causes heat stress and carbon monoxide (CO) buildup. Loading the C-130 aircraft by driving the vehicles onto the platform may expose the C-130 cabin crew to these environmental hazards. This study was aimed at investigating heat stress and CO exposure in the C-130 cabin during vehicle airlift. METHODS: There were four summer flights (two two vehicle, two three-vehicle; 2 d, 2 nights) studied. The cabin heat stress index (wet bulb globe temperature, WBGT) and CO levels before vehicle loading (control) were compared with those after vehicle loading. Furthermore, two- and three vehicle transportations, as well as day and night transportations, were compared. RESULTS: Ground temperature ranged from 18.2 to 33.4 degrees C. Mean heat stress index was higher in vehicle transportation than control flights, the greatest difference being 5.9 degrees C (p < 0.001). The WBGT levels exceeded the recommended exposure limit in 28 of 38 measurements during day flights. The cabin heat stress increased sharply with vehicle loading, and continued to increase for a range of 60-140 min after loading. Elevated cabin CO levels were found in three vehicle flights as compared with two, and in night flights as compared with day. CONCLUSIONS: In hot climates, C-130 vehicle transportation may exacerbate heat stress. The in-flight heat stress can be predicted by the ambient temperature, duration of the vehicle transportation, and number of transported vehicles. The cabin CO level is related to the number of transported vehicles. We recommend the use of effective environmental control systems during C-130 vehicle transportation in hot climates. PMID- 15828644 TI - Trans-femoral amputee pilots: criteria for return to the fighter cockpit. AB - Proximal lower limb amputations (trans-femoral) usually leave amputees with significant functional disturbances. This article contains information regarding three pilots with trans-femoral amputations that returned swiftly to continue their aeronautical careers despite their disabilities. Adaptations are needed in the limb prostheses to enable the amputees to access the minimally spaced cockpit. PMID- 15828645 TI - You're the flight surgeon. Myasthenia gravis. PMID- 15828646 TI - This month in aerospace medicine history--March 2005. PMID- 15828647 TI - This month in aerospace medicine history--April 2005. PMID- 15828648 TI - Effect of silhouette quality on hard problems in Gait recognition. AB - Gait as a behavioral biometric has been the subject of recent investigations. However, understanding the limits of gait-based recognition and the quantitative study of the factors effecting gait have been confounded by errors in the extracted silhouettes, upon which most recognition algorithms are based. To enable us to study this effect on a large population of subjects, we present a novel model based silhouette reconstruction strategy, based on a population based hidden Markov model (HMM), coupled with an eigen-stance model, to correct for common errors in silhouette detection arising from shadows and background subtraction. The model is trained and benchmarked using manually specified silhouettes for 71 subjects from the recently formulated HumanID Gait Challenge database. Unlike other essentially pixel-level silhouette cleaning methods, this method can remove shadows, especially between feet for the legs-apart stance, and remove parts due to any objects being carried, such as briefcase or a walking cane. After quantitatively establishing the improved quality of the silhouette over simple background subtraction, we show on the 122 subjects HumanID Gait Challenge Dataset and using two gait recognition algorithms that the observed poor performance of gait recognition for hard problems involving matching across factors such as surface, time, and shoe are not due to poor silhouette quality, beyond what is available from statistical background subtraction based methods. PMID- 15828650 TI - Adaptive critic autopilot design of bank-to-turn missiles using fuzzy basis function networks. AB - A new adaptive critic autopilot design for bank-to-turn missiles is presented. In this paper, the architecture of adaptive critic learning scheme contains a fuzzy basis-function-network based associative search element (ASE), which is employed to approximate nonlinear and complex functions of bank-to-turn missiles, and an adaptive critic element (ACE) generating the reinforcement signal to tune the associative search element. In the design of the adaptive critic autopilot, the control law receives signals from a fixed gain controller, an ASE and an adaptive robust element, which can eliminate approximation errors and disturbances. Traditional adaptive critic reinforcement learning is the problem faced by an agent that must learn behavior through trial-and-error interactions with a dynamic environment, however, the proposed tuning algorithm can significantly shorten the learning time by online tuning all parameters of fuzzy basis functions and weights of ASE and ACE. Moreover, the weight updating law derived from the Lyapunov stability theory is capable of guaranteeing both tracking performance and stability. Computer simulation results confirm the effectiveness of the proposed adaptive critic autopilot. PMID- 15828649 TI - A satisfactory-oriented approach to multiexpert decision-making with linguistic assessments. AB - This paper proposes a multiexpert decision-making (MEDM) method with linguistic assessments, making use of the notion of random preferences and a so-called satisfactory principle. It is well known that decision-making problems that manage preferences from different experts follow a common resolution scheme composed of two phases: an aggregation phase that combines the individual preferences to obtain a collective preference value for each alternative; and an exploitation phase that orders the collective preferences according to a given criterion, to select the best alternative/s. For our method, instead of using an aggregation operator to obtain a collective preference value, a random preference is defined for each alternative in the aggregation phase. Then, based on a satisfactory principle defined in this paper, that says that it is perfectly satisfactory to select an alternative as the best if its performance is as at least "good" as all the others under the same evaluation scheme, we propose a linguistic choice function to establish a rank ordering among the alternatives. Moreover, we also discuss how this linguistic decision rule can be applied to the MEDM problem in multigranular linguistic contexts. Two application examples taken from the literature are used to illuminate the proposed techniques. PMID- 15828651 TI - Multicriteria meta-heuristics for AGV dispatching control based on computational intelligence. AB - In many manufacturing environments, automated guided vehicles are used to move the processed materials between various pickup and delivery points. The assignment of vehicles to unit loads is a complex problem that is often solved in real-time with simple dispatching rules. This paper proposes an automated guided vehicles dispatching approach based on computational intelligence. We adopt a fuzzy multicriteria decision strategy to simultaneously take into account multiple aspects in every dispatching decision. Since the typical short-term view of dispatching rules is one of the main limitations of such real-time assignment heuristics, we also incorporate in the multicriteria algorithm a specific heuristic rule that takes into account the empty-vehicle travel on a longer time horizon. Moreover, we also adopt a genetic algorithm to tune the weights associated to each decision criteria in the global decision algorithm. The proposed approach is validated by means of a comparison with other dispatching rules, and with other recently proposed multicriteria dispatching strategies also based on computational Intelligence. The analysis of the results obtained by the proposed dispatching approach in both nominal and perturbed operating conditions (congestions, faults) confirms its effectiveness. PMID- 15828652 TI - An incremental approach to genetic-algorithms-based classification. AB - Incremental learning has been widely addressed in the machine learning literature to cope with learning tasks where the learning environment is ever changing or training samples become available over time. However, most research work explores incremental learning with statistical algorithms or neural networks, rather than evolutionary algorithms. The work in this paper employs genetic algorithms (GAs) as basic learning algorithms for incremental learning within one or more classifier agents in a multiagent environment. Four new approaches with different initialization schemes are proposed. They keep the old solutions and use an "integration" operation to integrate them with new elements to accommodate new attributes, while biased mutation and crossover operations are adopted to further evolve a reinforced solution. The simulation results on benchmark classification data sets show that the proposed approaches can deal with the arrival of new input attributes and integrate them with the original input space. It is also shown that the proposed approaches can be successfully used for incremental learning and improve classification rates as compared to the retraining GA. Possible applications for continuous incremental training and feature selection are also discussed. PMID- 15828653 TI - Boundary following and globally convergent path planning using instant goals. AB - In this paper, an Instant Goal approach is proposed for collision-free boundary following of obstacles of arbitrary shape and globally convergent path planning in unknown environments. Firstly, for effective knowledge representation and manipulation, a vector representation is presented, which not only saves much space but also conforms to the physical properties of range sensors. Secondly, the concept of Instant Goals is introduced enabling the robot to perform boundary following in a "natural" human-like manner, with additional measures taken to ensure that the robot is moving "forward" along the boundary, even if the obstacle is of arbitrary shape and disturbing obstacles are present. Collision checking is performed simultaneously and, when needed, collision avoidance is efficiently incorporated in. Based on the approach of boundary following, a realistic sensor-based path planner with global convergence property is designed for the robot capable of acquiring discrete and noisy range data. Realistic simulation experiments validate the effectiveness of the proposed approaches. PMID- 15828654 TI - Cooperative multiagent congestion control for high-speed networks. AB - An adaptive multiagent reinforcement learning method for solving congestion control problems on dynamic high-speed networks is presented. Traditional reactive congestion control selects a source rate in terms of the queue length restricted to a predefined threshold. However, the determination of congestion threshold and sending rate is difficult and inaccurate due to the propagation delay and the dynamic nature of the networks. A simple and robust cooperative multiagent congestion controller (CMCC), which consists of two subsystems: a long term policy evaluator, expectation-return predictor and a short-term rate selector composed of action-value evaluator and stochastic action selector elements has been proposed to solve the problem. After receiving cooperative reinforcement signals generated by a cooperative fuzzy reward evaluator using game theory, CMCC takes the best action to regulate source flow with the features of high throughput and low packet loss rate. By means of learning procedures, CMCC can learn to take correct actions adaptively under time-varying environments. Simulation results showed that the proposed approach can promote the system utilization and decrease packet losses simultaneously. PMID- 15828655 TI - Fuzzy combination of fuzzy and switching state-feedback controllers for nonlinear systems subject to parameter uncertainties. AB - This paper presents a fuzzy controller, which involves a fuzzy combination of local fuzzy and global switching state-feedback controllers, for nonlinear systems subject to parameter uncertainties with known bounds. The nonlinear system is represented by a fuzzy combined Takagi-Sugeno-Kang model, which is a fuzzy combination of the global and local fuzzy plant models. By combining the local fuzzy and global switching state-feedback controllers using fuzzy logic techniques, the advantages of both controllers can be retained and the undesirable chattering effect introduced by the global switching state-feedback controller can be eliminated. The steady-state error introduced by the global switching state-feedback controller when a saturation function is used can also be removed. Stability conditions, which are related to the system matrices of the local and global closed-loop systems, are derived to guarantee the closed-loop system stability. An application example will be given to demonstrate the merits of the proposed approach. PMID- 15828656 TI - Recognizing large isolated 3-D objects through next view planning using inner camera invariants. AB - Most model-based three-dimensional (3-D) object recognition systems use information from a single view of an object. However, a single view may not contain sufficient features to recognize it unambiguously. Further, two objects may have all views in common with respect to a given feature set, and may be distinguished only through a sequence of views. A further complication arises when in an image, we do not have a complete view of an object. This paper presents a new online scheme for the recognition and pose estimation of a large isolated 3-D object, which may not entirely fit in a camera's field of view. We consider an uncalibrated projective camera, and consider the case when the internal parameters of the camera may be varied either unintentionally, or on purpose. The scheme uses a probabilistic reasoning framework for recognition and next-view planning. We show results of successful recognition and pose estimation even in cases of a high degree of interpretation ambiguity associated with the initial view. PMID- 15828657 TI - Multiagent scheduling method with earliness and tardiness objectives in flexible job shops. AB - Flexible job-shop scheduling problems are an important extension of the classical job-shop scheduling problems and present additional complexity. Such problems are mainly due to the existence of a considerable amount of overlapping capacities with modern machines. Classical scheduling methods are generally incapable of addressing such capacity overlapping. We propose a multiagent scheduling method with job earliness and tardiness objectives in a flexible job-shop environment. The earliness and tardiness objectives are consistent with the just-in-time production philosophy which has attracted significant attention in both industry and academic community. A new job-routing and sequencing mechanism is proposed. In this mechanism, two kinds of jobs are defined to distinguish jobs with one operation left from jobs with more than one operation left. Different criteria are proposed to route these two kinds of jobs. Job sequencing enables to hold a job that may be completed too early. Two heuristic algorithms for job sequencing are developed to deal with these two kinds of jobs. The computational experiments show that the proposed multiagent scheduling method significantly outperforms the existing scheduling methods in the literature. In addition, the proposed method is quite fast. In fact, the simulation time to find a complete schedule with over 2000 jobs on ten machines is less than 1.5 min. PMID- 15828658 TI - Hierarchical Kohonenen net for anomaly detection in network security. AB - A novel multilevel hierarchical Kohonen Net (K-Map) for an intrusion detection system is presented. Each level of the hierarchical map is modeled as a simple winner-take-all K-Map. One significant advantage of this multilevel hierarchical K-Map is its computational efficiency. Unlike other statistical anomaly detection methods such as nearest neighbor approach, K-means clustering or probabilistic analysis that employ distance computation in the feature space to identify the outliers, our approach does not involve costly point-to-point computation in organizing the data into clusters. Another advantage is the reduced network size. We use the classification capability of the K-Map on selected dimensions of data set in detecting anomalies. Randomly selected subsets that contain both attacks and normal records from the KDD Cup 1999 benchmark data are used to train the hierarchical net. We use a confidence measure to label the clusters. Then we use the test set from the same KDD Cup 1999 benchmark to test the hierarchical net. We show that a hierarchical K-Map in which each layer operates on a small subset of the feature space is superior to a single-layer K-Map operating on the whole feature space in detecting a variety of attacks in terms of detection rate as well as false positive rate. PMID- 15828659 TI - Autonomous stair-climbing with miniature jumping robots. AB - The problem of vision-guided control of miniature mobile robots is investigated. Untethered mobile robots with small physical dimensions of around 10 cm or less do not permit powerful onboard computers because of size and power constraints. These challenges have, in the past, reduced the functionality of such devices to that of a complex remote control vehicle with fancy sensors. With the help of a computationally more powerful entity such as a larger companion robot, the control loop can be closed. Using the miniature robot's video transmission or that of an observer to localize it in the world, control commands can be computed and relayed to the inept robot. The result is a system that exhibits autonomous capabilities. The framework presented here solves the problem of climbing stairs with the miniature Scout robot. The robot's unique locomotion mode, the jump, is employed to hop one step at a time. Methods for externally tracking the Scout are developed. A large number of real-world experiments are conducted and the results discussed. PMID- 15828660 TI - Fuzzy OLAP association rules mining-based modular reinforcement learning approach for multiagent systems. AB - Multiagent systems and data mining have recently attracted considerable attention in the field of computing. Reinforcement learning is the most commonly used learning process for multiagent systems. However, it still has some drawbacks, including modeling other learning agents present in the domain as part of the state of the environment, and some states are experienced much less than others, or some state-action pairs are never visited during the learning phase. Further, before completing the learning process, an agent cannot exhibit a certain behavior in some states that may be experienced sufficiently. In this study, we propose a novel multiagent learning approach to handle these problems. Our approach is based on utilizing the mining process for modular cooperative learning systems. It incorporates fuzziness and online analytical processing (OLAP) based mining to effectively process the information reported by agents. First, we describe a fuzzy data cube OLAP architecture which facilitates effective storage and processing of the state information reported by agents. This way, the action of the other agent, not even in the visual environment. of the agent under consideration, can simply be predicted by extracting online association rules, a well-known data mining technique, from the constructed data cube. Second, we present a new action selection model, which is also based on association rules mining. Finally, we generalize not sufficiently experienced states, by mining multilevel association rules from the proposed fuzzy data cube. Experimental results obtained on two different versions of a well-known pursuit domain show the robustness and effectiveness of the proposed fuzzy OLAP mining based modular learning approach. Finally, we tested the scalability of the approach presented in this paper and compared it with our previous work on modular-fuzzy Q-learning and ordinary Q-learning. PMID- 15828661 TI - Identifying critical variables of principal components for unsupervised feature selection. AB - Principal components analysis (PCA) is probably the best-known approach to unsupervised dimensionality reduction. However, axes of the lower-dimensional space, ie., principal components (PCs), are a set of new variables carrying no clear physical meanings. Thus, interpretation of results obtained in the lower dimensional PCA space and data acquisition for test samples still involve all of the original measurements. To deal with this problem, we develop two algorithms to link the physically meaningless PCs back to a subset of original measurements. The main idea of the algorithms is to evaluate and select feature subsets based on their capacities to reproduce sample projections on principal axes. The strength of the new algorithms is that the computaion complexity involved is significantly reduced, compared with the data structural similarity-based feature evaluation. PMID- 15828662 TI - Performance enhancement of a contract net protocol based system through instance based learning. PMID- 15828663 TI - Hybridization of fuzzy GBML approaches for pattern classification problems. PMID- 15828664 TI - Supervisory control of fuzzy discrete event systems. PMID- 15828665 TI - Designing stable MIMO fuzzy controllers. PMID- 15828666 TI - Second-order training of adaptive critics for online process control. PMID- 15828667 TI - A pilot study of mercury liberation and capture from coal-fired power plant fly ash. AB - The coal-fired electric utility generation industry has been identified as the largest anthropogenic source of mercury (Hg) emissions in the United States. One of the promising techniques for Hg removal from flue gas is activated carbon injection (ACI). The aim of this project was to liberate Hg bound to fly ash and activated carbon after ACI and provide high-quality coal combustion products for use in construction materials. Both bench- and pilot-scale tests were conducted to liberate Hg using a thermal desorption process. The results indicated that up to 90% of the Hg could be liberated from the fly ash or fly-ash-and-activated carbon mixture using a pilot-scale apparatus (air slide) at 538 degrees C with a very short retention time (less than 1 min). Scanning electron microscope (SEM) evaluation indicated no significant change in fly ash carbon particle morphology following the thermal treatment. Fly ash particles collected in the baghouse of the pilot-scale apparatus were smaller in size than those collected at the exit of the air slide. A similar trend was observed in carbon particles separated from the fly ash using froth flotation. The results of this study suggest a means for power plants to reduce the level of Hg in coal-combustion products and potentially recycle activated carbon while maintaining the resale value of fly ash. This technology is in the process of being patented. PMID- 15828668 TI - Effects of SO2 and NOx emission reductions on PM2.5 mass concentrations in the southeastern United States. AB - Measurements from sites of the Southeastern Aerosol Research and Characterization (SEARCH) program, made from 1998 to 2001, are used with a thermodynamic equilibrium model, Simulating Composition of Atmospheric Particles at Equilbrium (SCAPE2), to extend an earlier investigation of the responses of fine particulate nitrate (NO3-) and fine particulate matter (PM2.5) mass concentrations to changes in concentrations of nitric acid (HNO3) and sulfate (SO42-). The responses were determined for a projected range of variations of SO42- and HNO3 concentrations resulting from adopted and proposed regulatory initiatives. The predicted PM2.5 mass concentration decreases averaged 1.8-3.9 microg/m3 for SO42- decreases of 46 63% from current concentrations. Combining the S042- decrease with a 40% HNO3 decrease from current concentrations (approximating expected mobile-source oxides of nitrogen [NOx] reductions by 2020) yielded additional incremental reductions of mean predicted PM2.5 mass concentration of 0.2 microg/m3 for three nonurban sites and 0.8-1 microg/m3 for one nonurban and two urban sites. Increasing the HNO3 reduction to 55% (an estimate of adding Clear Skies Phase II NOx reductions) yielded additional incremental reductions of mean predicted PM2.5 mass concentration of 0-0.4 microg/m3. Because of the well-documented losses of particulate NO3- from Federal Reference Method (FRM) filters, only a fraction of these incremental changes would be observed. PMID- 15828669 TI - An investigation of distributed lag models in the context of air pollution and mortality time series analysis. AB - In particulate air pollution mortality time series studies, the particulate air pollution exposure measure used is typically the current day's or the previous day's air pollution concentration or a multi-day moving average air pollution concentration. Distributed lag models (DLMs) that allow for differential air pollution effects that are spread over multiple days are seen as an improvement over using a single- or multi-day moving average air pollution exposure measure. However, at the current time, the statistical properties of DLMs as a measure of air pollution exposure have not been investigated. In this paper, a simulation study is used to investigate the performance of DLMs as a measure of air pollution exposure in comparison with single- and multi-day moving average air pollution exposure measures under various forms for the true effect of air pollution on mortality. The simulation study shows that DLMs offer a more robust measure of the effect of air pollution on mortality and avoid the potential for a large negative bias compared with single- or multi-day moving average air pollution exposure measures. This is important information. In many U.S. cities, particulate air pollution concentrations are observed only once every six days, meaning it is often only possible to use single-day particulate air pollution exposure measures. The results from this paper will help quantify the magnitude of the negative bias that can result from using single-day exposure measures. The implications of this work for future air pollution mortality time series studies are discussed. The data used in this paper are concurrent daily time series of mortality, weather, and particulate air pollution from Cook County, IL, for the period 1987-1994. PMID- 15828670 TI - Compressibility of municipal solid waste codisposed with fly ash. AB - If a municipal solid waste (MSW) landfill is used as the foundation for a construction site, the change in the loading conditions may cause considerable compression of the landfill. Therefore, reinforcement to compensate for the loose compression nature of a MSW landfill is a very important design factor for geotechnical engineers when considering the end-use of the landfill. In this study, we discuss a possible technique for stabilizing MSW landfills through the codisposal of municipal solid waste and fly ash. To estimate the stabilization based on the compression characteristics of the codisposed landfill, we performed tests using a large compression set and experimental cells. According to the test results, if the proportion of fly ash is increased, initial and long-term compression could be reduced. PMID- 15828671 TI - Seasonal and spatial variation of solvent extractable organic compounds in fine suspended particulate matter in Hong Kong. AB - The results of a 12-month study of more than 100 solvent extractable organic compounds (SEOC) in particulate matter (PM) less than or equal to 2.5 microm (PM2.5) collected at three air monitoring stations located at roadside, urban, and rural sites in Hong Kong are reported. The total yield of SEOC that accounts for approximately 8-18% of organic carbon (OC) determined by a thermal optical transmittance method was 125-2060 ng/m3, which included 14.6-128 ng/m3 resolved aliphatic hydrocarbons, 39.4-1380 ng/m3 unresolved complex mixtures, 0.6-17.2 ng/m3 polycyclic aromatic hydrocarbons, 41.6-520 ng/m3 fatty acids, and < 0.1 12.1 ng/m3 alkanols. Distinct seasonal variations (summer/winter differences) were observed with higher concentrations of the total and each class of SEOC in the winter and lower concentrations in the summer. Spatial variations are also obvious, with the roadside samples having the highest concentrations of SEOC and the rural samples having the lowest concentrations in all seasons. Characteristic ratios of petroleum hydrocarbons, such as carbon preference index, unresolved to resolved components, and carbon number with maximum concentration, suggest that PM2.5 carbon in Hong Kong originates from both biogenic and anthropogenic sources. The proportion of SEOC in PM2.5 from anthropogenic sources is estimated. PMID- 15828672 TI - Complying with Executive Order 13148 using the Enterprise Environmental Safety And Occupational Health Management Information System. AB - All U.S. Department of Defense (DoD) facilities are required under Executive Order (EO) 13148, "Greening the Government through Leadership in Environmental Management," to establish quality-based environmental management systems (EMSs) that support environmental decision-making and verification of continuous environmental improvement by December 31, 2005. Compliance with EO 13148 as well as other federal, state, and local environmental regulations places a significant information management burden on DoD facilities. Cost-effective management of environmental data compels DoD facilities to establish robust database systems that not only address the complex and multifaceted environmental monitoring, record-keeping, and reporting requirements demanded by these rules but enable environmental management decision-makers to gauge improvements in environmental performance. The Enterprise Environmental Safety and Occupational Health Management Information System (EESOH-MIS) is a new electronic database developed by the U.S. Air Force to manage both the data needs associated with regulatory compliance programs across its facilities as well as the non-regulatory environmental information that supports installation business practices. The U.S. Air Force, which has adopted the Plan-Do-Check-Act methodology as the EMS standard that it will employ to address EO 13148 requirements. PMID- 15828673 TI - Nickel and sulfur speciation of residual oil fly ashes from two electric utility steam-generating units. AB - Representative duplicate fly ash samples were obtained from the stacks of 400- and 385-MW utility boilers (Unit A and Unit B, respectively) using a modified U.S. Environmental Protection Agency (EPA) Method 17 sampling train assembly as they burned 0.9 and 0.3 wt % S residual (No. 6 fuel) oils, respectively, during routine power plant operations. Residual oil fly ash (ROFA) samples were analyzed for Ni concentrations and speciation using inductively coupled plasma-atomic emission spectroscopy, X-ray absorption fine structure (XAFS) spectroscopy, and X ray diffraction (XRD). ROFA deionized H2O extraction residues were also analyzed for Ni speciation using XAFS and XRD. Total Ni concentrations in the ROFAs were similar, ranging from 1.3-1.5 wt%; however, stack gas Ni concentrations in the Unit A were 0.990 microg/Nm3 compared with 0.620 microg/Nm3 for Unit B because of the greater residual oil feed rates employed at Unit A to attain higher 400-MW load conditions with a lower heating value oil. Ni speciation analysis results indicated that ROFAs from Unit A contain approximately 3 wt % NiSO4 x xH2O (where x is assumed to be 6 for calculation purposes) and appoximately 4.5 wt% of a Ni containing spinel compound, similar in composition to (Mg,Ni)(Al,Fe)2O4. ROFAs from Unit B contain on average 2 wt% NiSO4 x 6 H20 and 1.1 wt% NiO. XAFS and XRD analyses did not detect any nickel sulfide compounds, including carcinogenic nickel subsulfide (Ni3S2) (XAFS detection limit is 5% of the total Ni concentration). In addition, XAFS measurements indicated that inorganic sulfate and organic thiophene species accounted for > 97% of the total S in the ROFAs. Unit A ROFAs contained much lower thiophene proportions because cyclone-separated ROFA reinjection is employed on this unit to collect and reburn the larger carbonaceous particles. PMID- 15828674 TI - An efficient venturi scrubber system to remove submicron particles in exhaust gas. AB - An efficient venturi scrubber system making use of heterogeneous nucleation and condensational growth of particles was designed and tested to remove fine particles from the exhaust of a local scrubber where residual SiH4 gas was abated and lots of fine SiO2 particles were generated. In front of the venturi scrubber, normal-temperature fine-water mist mixes with high-temperature exhaust gas to cool it to the saturation temperature, allowing submicron particles to grow into micron sizes. The grown particles are then scrubbed efficiently in the venturi scrubber. Test results show that the present venturi scrubber system is effective for removing submicron particles. For SiO2 particles greater than 0.1microm, the removal efficiency is greater than 80-90%, depending on particle concentration. The corresponding pressure drop is relatively low. For example, the pressure drop of the venturi scrubber is approximately 15.4 +/- 2.4 cm H2O when the liquid-to gas ratio is 1.50 L/m3. A theoretical calculation has been conducted to simulate particle growth process and the removal efficiency of the venturi scrubber. The theoretical results agree with the experimental data reasonably well when SiO2 particle diameter is greater than 0.1 microm. PMID- 15828675 TI - Seasonal source-receptor relationships in a petrochemical industrial district over northern Taiwan. AB - This study investigated the relationships between meteorological data, pollution sources, and receptors over northern Taiwan. During the intensive sampling period in summer 1992, the weather was controlled predominantly by a Pacific subtropical high and by Typhoon Mark. During the other intensive sampling period in winter 1993, while a cold frontal system approached Taiwan, the northeasterly winds prevailed most of the time. The local circulation such as land-sea breeze only developed under weak synoptic environment. Particle concentrations and element composition in winter were higher than in summer. This can be attributed to the high convection of air mass, which leads to the vertical dispersion of pollutants in summer. In addition to the subtropical high pressure, typhoons are frequently accompanied with high-wind speeds and unstable weather conditions that also dilute and eliminate the pollutants. In winter, the prevailing northeasterlies might carry pollutants from Midland China. Furthermore, the anticyclone system develops a stagnant condition that easily leads to pollutant accumulation. In this case, the wind direction affected the source contribution of the receptor and the PM10 displays a higher correlation with coarse and fine particulate than meteorological parameters in summer. In addition, the mixing height shows a high correlation with PM10 in winter. PMID- 15828676 TI - Analysis of PM10, PM2.5, and PM2 5-10 concentrations in Santiago, Chile, from 1989 to 2001. AB - Daily particle samples were collected in Santiago, Chile, at four urban locations from January 1, 1989, through December 31, 2001. Both fine PM with da < 2.5 microm (PM2.5) and coarse PM with 2.5 < da < 10 microm (PM2.5-10) were collected using dichotomous samplers. The inhalable particle fraction, PM10, was determined as the sum of fine and coarse concentrations. Wind speed, temperature and relative humidity (RH) were also measured continuously. Average concentrations of PM2.5 for the 1989-2001 period ranged from 38.5 microg/m3 to 53 microg/m3. For PM2.5-10 levels ranged from 35.8-48.2 microg/m3 and for PM10 results were 74.4 101.2 microg/m3 across the four sites. Both annual and daily PM2.5 and PM10 concentration levels exceeded the U.S. National Ambient Air Quality Standards and the European Union concentration limits. Mean PM2.5 levels during the cold season (April through September) were more than twice as high as those observed in the warm season (October through March); whereas coarse particle levels were similar in both seasons. PM concentration trends were investigated using regression models, controlling for site, weekday, month, wind speed, temperature, and RH. Results showed that PM2.5 concentrations decreased substantially, 52% over the 12 year period (1989-2000), whereas PM2.5-10 concentrations increased by approximately 50% in the first 5 years and then decreased by a similar percentage over the following 7 years. These decreases were evident even after controlling for significant climatic effects. These results suggest that the pollution reduction programs developed and implemented by the Comision Nacional del Medio Ambiente (CONAMA) have been effective in reducing particle levels in the Santiago Metropolitan region. However, particle levels remain high and it is thus imperative that efforts to improve air quality continue. PMID- 15828677 TI - PM source identification at Sunland Park, New Mexico, using a simple heuristic meteorological and chemical analysis. AB - The causes for evening low-wind PM10 and PM2.5 peaks at Sunland Park, NM, were investigated by using wind sector analysis and by assessing relationships between PM loadings and meteorological parameters through canonical ordination analysis. Both PM10 and PM2.5 concentrations during the evening hours accounted for approximately 50% of their respective 24-hr averages, and the PM10 was mainly composed of coarse material (PM10-2.5 amounted to 77% of PM10). A wind sector analysis based on data from three surface meteorological monitoring stations in the region narrowed the potential source region for PM10 and PM2.5 to an area within a few kilometers south of Sunland Park. Canonical ordination analysis confirmed that the peak frequently occurred under stable conditions with weak southerly winds. Chemical analyses of PM showed that elemental and organic carbon (EC and OC, respectively) dominate PM2.5 and inorganic elements dominate PM10 2.5. The combined data for EC/OC, geologic elements, and various trace elements indicate that under low wind and stable conditions, traffic-related PM emissions (motor vehicle exhausts and re-suspended road dust) from the south of the site are the most likely sources for the evening PM10 and PM2.5 peaks. PMID- 15828678 TI - Maintaining high-volume, low-pressure surface-coating regulatory compliance using the U.s. Environmental Protection Agency's data quality objectives process. AB - To effectively reduce the environmental compliance costs associated with meeting specific requirements under the Aerospace Manufacturing and Rework Facility's National Emission Standard for Hazardous Air Pollutants rule, the U.S. Environmental Protection Agency's (EPA) Data Quality Objective (DQO) process has been proposed as a suitable framework for developing a scientifically defensible surface compliance monitoring program. By estimating the variability associated with the air cap pressure of high- volume, low-pressure (HVLP) surface-coating spray equipment, the number of monitoring samples necessary for an affected facility to claim compliance with a desired statistical confidence level was established. Using data taken from the pilot test facility, the DQO process indicated that the mean of at least 21 HVLP air cap pressure samples taken over the compliance period must be < or = 10 pounds per square inch (psig) gauge for the facility to claim regulatory compliance with 99.99% statistical confidence. Fewer compliance samples could be taken, but that decision would lead to a commensurate reduction in the compliance confidence level. Implementation of the DQO-based compliance sampling plan eliminates the need for an affected facility to sample all regulated HVLP surface-coating processes while still maintaining a high level of compliance assurance. PMID- 15828679 TI - Proposed guidelines for a unified nomenclature and phylogenetic analysis of type III Hop effector proteins in the plant pathogen Pseudomonas syringae. AB - Pathovars of Pseudomonas syringae interact with their plant hosts via the action of Hrp outer protein (Hop) effector proteins, injected into plant cells by the type III secretion system (TTSS). Recent availability of complete genome sequences for a number of P. syringae pathovars has led to a significant increase in the rate of effector discovery. However, lack of a systematic nomenclature has resulted in multiple names being assigned to the same Hop, unrelated Hops designated by the same alphabetic character, and failure of name choices to reflect consistent standards of experimental confirmation or phylogenetic relatedness. Therefore, specific experimental and bioinformatic criteria are proposed for proteins to be designated as Hops. A generic Hop name structure, HopXY#pv strain, also is proposed, wherein family membership is indicated by the alphabetic characters, subgroup membership numerically, and source pathovar and strain in subscript. Guidelines are provided for phylogenetic characterization and name selection for Hops that are novel, related to previously characterized Hops, chimeras, pseudogenes, truncations, or nonexpressed alleles. Phylogenetic analyses of previously characterized Hops are described, the results of which have been used to guide their integration into the proposed nomenclature. PMID- 15828680 TI - A new cell-to-cell transport model for Potexviruses. AB - In the last five years, we have gained significant insight into the role of the Potexvirus proteins in virus movement and RNA silencing. Potexviruses require three movement proteins, named triple gene block (TGB)p1, TGBp2, and TGBp3, and the viral coat protein (CP) to facilitate viral cell-to-cell and vascular transport. TGBp1 is a multifunctional protein that has RNA helicase activity, promotes translation of viral RNAs, increases plasmodesmal size exclusion limits, and suppresses RNA silencing. TGBp2 and TGBp3 are membrane-binding proteins. CP is required for genome encapsidation and forms ribonucleoprotein complexes along with TGBp1 and viral RNA. This review considers the functions of the TGB proteins, how they interact with each other and CP, and how silencing suppression might be linked to viral transport. A new model of the mechanism for Potexvirus transport is proposed. PMID- 15828681 TI - Barley Rom1 reveals a potential link between race-specific and nonhost resistance responses to powdery mildew fungi. AB - The Rar1 gene, identified in the context of race-specific powdery mildew resistance mediated by the Hordeum vulgare (barley) resistance (R) gene Mla12, is required for the function of many R-mediated defense responses in mono- and dicotyledonous plant species. Mla resistance is associated with an oxidative burst and a subsequent cell death reaction of attacked cells. Rar1 mutants are impaired in these responses and, to identify genetic elements which negatively regulate the Mla12-triggered response, we have screened mutagenized Mla12 rar1 mutant populations for restoration of the resistance response. Here we describe the restoration of Mla12-specified resistance (rom1) mutant that restores features of disease resistance to a Blumeria graminis f. sp. hordei isolate expressing the avirulence gene AvrMla12 and retains susceptibility to an isolate lacking AvrMla12. Histochemical analyses show that, in rom1 mutant plants, a whole-cell oxidative burst and cell death response in attacked epidermal cells is restored in the incompatible interaction. Defense responses against tested inappropriate powdery mildews, B. graminis f. sp. tritici and Golovinomyces orontii, were diminished in rar1 mutant plants and enhanced in rom1 mutant plants relative to the wild type. These findings indicate antagonistic activities of Rar1 and Rom1 and reveal their contribution to nonhost and race-specific resistance responses. PMID- 15828682 TI - Molecular properties of the Xanthomonas AvrRxv effector and global transcriptional changes determined by its expression in resistant tomato plants. AB - The Xanthomonas campestris pv. vesicatoria avirulence gene AvrRxv specifies resistance on the tomato line Hawaii 7998 by interacting with three nondominant plant resistance genes. AvrRxv molecular properties that impinge on its avirulence activity were characterized and transcriptional changes caused by AvrRxv expression in resistant tomato plants were extensively examined. AvrRxv localized predominantly to the cytoplasm and possibly in association with plasma and nuclear membranes in both resistant and susceptible tomato plants. The AvrRxv cysteine protease catalytic core was found to be essential for host recognition, because introduction of mutations in this domain affected the ability of AvrRxv to elicit a hypersensitive response and the inhibition of bacterial growth in resistant plants. In addition, expression profiles were analyzed for approximately 8,600 tomato genes in resistant plants challenged with X. campestris pv. vesicatoria strains expressing wild-type AvrRxv or a catalytic core AvrRxv mutant. In all, 420 genes were identified as differentially modulated by the expression of a functional AvrRxv, including over 15 functional classes of proteins and a large number of transcription factors and signaling components. Findings of this study allow the development of new hypotheses about the molecular basis of recognition between AvrRxv and the corresponding resistance proteins, and set the stage for the dissection of signaling and cellular responses triggered in tomato plants by this avirulence factor. PMID- 15828683 TI - Transcriptional analysis of the Azospirillum brasilense indole-3-pyruvate decarboxylase gene and identification of a cis-acting sequence involved in auxin responsive expression. AB - Expression of the Azospirillum brasilense ipdC gene, encoding an indole-3 pyruvate decarboxylase, a key enzyme in the production of indole-3-acetic acid (IAA) in this bacterium, is upregulated by IAA. Here, we demonstrate that the ipdC gene is the promoter proximal gene in a bicistronic operon. Database searches revealed that the second gene of this operon, named iaaC, is well conserved evolutionarily and that the encoded protein is homologous to the Escherichia coli protein SCRP-27A, the zebrafish protein ES1, and the human protein KNP-I/GT335 (HES1), all of unknown function and belonging to the DJ 1/PfpI superfamily. In addition to this operon structure, iaaC is also transcribed monocistronically. Mutation analysis of the latter gene indicated that the encoded protein is involved in controlling IAA biosynthesis but not ipdC expression. Besides being upregulated by IAA, expression of the ipdC-iaaC operon is pH dependent and maximal at acidic pH. The ipdC promoter was studied using a combination of deletion analyses and site-directed mutagenesis. A dyadic sequence (ATTGTTTC(GAAT)GAAACAAT), centered at -48 was demonstrated to be responsible for the IAA inducibility. This bacterial auxin-responsive element does not control the pH-dependent expression of ipdC-iaaC. PMID- 15828684 TI - Oligonucleotide microarray analysis of the salA regulon controlling phytotoxin production by Pseudomonas syringae pv. syringae. AB - The salA gene is a key regulatory element for syringomycin production by Pseudomonas syringae pv. syringae and encodes a member of the LuxR regulatory protein family. Previous studies revealed that salA, a member of the GacS/GacA signal transduction system, was required for bacterial virulence, syringomycin production, and expression of the syrB1 synthetase gene. To define the SalA regulon, the spotted oligonucleotide microarray was constructed using gene specific 70-mer oligonucleotides of all open reading frames (ORFs) predicted in the syringomycin (syr) and syringopeptin (syp) gene clusters along with representative genes important to bacterial virulence, growth, and survival. The microarray containing 95 oligos was used to analyze transcriptional changes in a salA mutant (B301DSL07) and its wild-type strain, B301D. Expression of 16 genes was significantly higher (> twofold) in B301D than in the salA mutant; the maximum change in expression was 15-fold for some toxin biosynthesis genes. Except for the sylD synthetase gene for syringolin production, all ORFs controlled by SalA were located in the syr-syp genomic island and were associated with biosynthesis, secretion, and regulation of syringomycin and syringopeptin. The positive regulatory effect of SalA on transcription of sypA, syrB1, syrC, and sylD was verified by reporter fusions or real-time polymerase chain reaction analysis. None of the genes or ORFs was significantly down-regulated by the salA gene. These results demonstrated that a subgenomic oligonucleotide microarray is a powerful tool for defining the SalA regulon and its relationship to other genes important to plant pathogenesis. PMID- 15828685 TI - Identification of a new quorum-sensing-controlled virulence factor in Erwinia carotovora subsp. atroseptica secreted via the type II targeting pathway. AB - Two-dimensional polyacrylamide gel electrophoresis of the secreted proteins of Erwinia carotovora subsp. atroseptica revealed a low-abundance protein that was identified by mass spectrometry as a homologue of a Xanthomonas campestris avirulence protein with unknown function. The predicted Svx protein has an N terminal signal sequence and zinc binding-region signature, and the mature protein is post-translationally modified. A 2D difference gel electrophoresis (DIGE) showed that the protein is secreted by the type II (out) secretion apparatus, which is also responsible for the secretion of the major known virulence factors, PelC and CelV. Transcription of the svx gene is under N-acyl homoserine lactone-mediated quorum-sensing control. The svx gene was inactivated by transposon insertion. The mutant showed a decrease in virulence in potato plant assays, demonstrating a role for Svx in the pathogenicity of E. carotovora subsp. atroseptica. These results show that Svx is a previously unidentified virulence determinant which is secreted by the out machinery and is regulated by quorum sensing, two systems employed by several other virulence factors. Thus, the type II secretory machine is a conduit for virulence factors other than the main pectinnases and cellulase in E. carotovora subsp. atroseptica. PMID- 15828686 TI - Novel quorum-sensing-controlled genes in Erwinia carotovora subsp. carotovora: identification of a fungal elicitor homologue in a soft-rotting bacterium. AB - Seven new genes controlled by the quorum-sensing signal molecule N-(3 oxohexanoyl)-L-homoserine lactone (OHHL) have been identified in Erwinia carotovora subsp. carotovora. Using TnphoA as a mutagen, we enriched for mutants defective in proteins that could play a role in the interaction between E. carotovora subsp. carotovora and its plant hosts, and identified NipEcc and its counterpart in E. carotovora subsp. atroseptica. These are members of a growing family of proteins related to Nep1 from Fusarium oxysporum which can induce necrotic responses in a variety of dicotyledonous plants. NipEcc produced necrosis in tobacco, NipEca affected potato stem rot, and both affected virulence in potato tubers. In E. carotovora subsp. carotovora, nip was shown to be subject to weak repression by the LuxR family regulator, EccR, and may be regulated by the negative global regulator RsmA. PMID- 15828687 TI - Tomato defense to Oidium neolycopersici: dominant Ol genes confer isolate dependent resistance via a different mechanism than recessive ol-2. AB - Tomato powdery mildew caused by Oidium neolycopersici has become a globally important disease of tomato (Lycopersicon esculentum). To study the defense responses of tomato triggered by tomato powdery mildew, we first mapped a set of resistance genes to O. neolycopersici from related Lycopersicon species. An integrated genetic map was generated showing that all the dominant resistance genes (Ol-1, Ol-3, Ol-4, Ol-5, and Ol-6) are located on tomato chromosome 6 and are organized in three genetic loci. Then, near-isogenic lines (NIL) were produced that contain the different dominant Ol genes in a L. esculentum genetic background. These NIL were used in disease tests with local isolates of O. neolycopersici in different geographic locations, demonstrating that the resistance conferred by different Ol genes was isolate-dependent and, hence, may be race-specific. In addition, the resistance mechanism was analyzed histologically. The mechanism of resistance conferred by the dominant Ol genes was associated with hypersensitive response, which varies in details depending on the Ol-gene in the NIL, while the mechanism of resistance governed by the recessive gene ol-2 on tomato chromosome 4 was associated with papillae formation. PMID- 15828688 TI - Arabidopsis ssi2-conferred susceptibility to Botrytis cinerea is dependent on EDS5 and PAD4. AB - Loss of a stearoyl-ACP desaturase activity in the Arabidopsis thaliana ssi2 mutant confers susceptibility to the necrotroph, Botrytis cinerea. In contrast, the ssi2 mutant exhibits enhanced resistance to Pseudomonas syringae, Peronospora parasitica, and Cucumber mosaic virus. The altered basal resistance to these pathogens in the ssi2 mutant plant is accompanied by the constitutive accumulation of elevated salicylic acid (SA) level and expression of the pathogenesis-related 1 (PR1) gene, the inability of jasmonic acid (JA) to activate expression of the defensin gene, PDF1.2, and the spontaneous death of cells. Here, we show that presence of the eds5 and pad4 mutant alleles compromises the ssi2-conferred resistance to Pseudomonas syringae pv. maculicola. In contrast, resistance to B. cinerea was restored in the ssi2 eds5 and ssi2 pad4 double-mutant plants. However, resistance to B. cinerea was not accompanied by the restoration of JA responsiveness in the ssi2 eds5 and ssi2 pad4 plants. The ssi2 eds5 and ssi2 pad4 plants retain the ssi2-conferred spontaneous cell death phenotype, suggesting that cell death is not a major factor that predisposes the ssi2 mutant to infection by B. cinerea. Furthermore, the high SA content of the ssi2 pad4 plant, combined with our previous observation that the SA-deficient ssi2 nahG plant succumbs to infection by B. cinerea, suggests that elevated SA level does not have a causal role in the ssi2-conferred susceptibility to B. cinerea. Our results suggest that interaction between an SSI2-dependent factor or factors and an EDS5- and PAD4-dependent mechanism or mechanisms modulates defense to B. cinerea. PMID- 15828689 TI - Medical management of Iraqi enemy prisoners of war during Operation Telic. AB - OBJECTIVE: To describe the background to the provision of medical support to enemy prisoners of war (EPW) and the clinical activity undertaken at the EPW medical treatment facility. METHODS: Data were recovered from operational diaries and individual patient records to show EPW attendance rates at routine sick call, diagnoses made, and medications prescribed. RESULTS: EPW presented at an average rate of 1.2 cases per 100 EPW per day. Trauma, musculoskeletal, and dental problems accounted for 52% of presentations. Medications for mild/moderate pain and broad-spectrum antibiotics accounted for nearly 65% of prescriptions. Cultural and security issues presented additional challenges to medical staff members. CONCLUSIONS: Provision of medical support for EPW is an enduring task covered by the Geneva Conventions. EPW present a wide range of ailments at routine sick call that require suitable medications, all provided in a culturally appropriate and secure environment. PMID- 15828690 TI - An assessment of health status among medical research volunteers who served in the Project Whitecoat program at Fort Detrick, Maryland. AB - Between 1954 and 1973, more than 2000 men entering military service as conscientious objectors participated in Project Whitecoat as medical research volunteers for the Army's biological warfare defense program. An assessment of self-reported, current health status among 358 "exposed" individuals and 164 unexposed control subjects found no conclusive evidence that receipt of investigational agents was related to adverse health outcomes. No differences in current overall health, current exercise levels, self-reported symptoms, and self reported medical conditions were seen between the study groups. Possible associations were seen between exposure to antibiotics or other biological agents and self-reported asthma (13.0% vs. 2.4%, relative risk [RR] = 6.00, 95% confidence interval [CI] = 1.03-34.90, p = 0.050), as well as between receipt of tularemia vaccine(s) and self-reported asthma (13.3% vs. 2.4%, RR = 6.15, 95% CI = 1.03-36.70, p = 0.049) and increased frequency/severity of headaches (35.6% vs. 18.3%, RR = 2.46, 95% CI = 0.99-6.15, p = 0.074). However, the size of the population under study was insufficient to assert with confidence that these statistical associations are real. PMID- 15828691 TI - Military nursing research by students at the Graduate School of Nursing Uniformed Services University of the Health Sciences. AB - Military nursing research has had a long and productive history. Today, much of this research is conducted under two programs, the TriService Nursing Research Program and the Graduate School of Nursing (GSN), both located at the Uniformed Services University of the Health Sciences in Bethesda, Maryland. This article will discuss the 150 military nursing research projects carried out by students at the GSN since its founding in 1992. Although most projects have been small in scope, they have obtained useful results. Some projects have served as the basis for larger-scale research studies, receiving funding from the TriService Nursing Research Program. Reports of all projects are available in an online database and some have been published in professional journals. This review concludes that the research produced by GSN students has been beneficial to students and to the military health system. PMID- 15828692 TI - Malaria on a military peacekeeping operation: a case study with no cases. AB - BACKGROUND: Malaria continues to be a disease of importance to travelers and the military is no exception. Individual protection measures based on advice, bite avoidance, chemoprophylaxis, and diagnosis are advocated for protection against the disease. However, the military has an additional strand to malaria protection -the chain of command. AIM: To describe the experience of a British military deployment where the Force Commander took a proactive approach to malaria protection. RESULTS: In 512 person-weeks of exposure in a theater with high rates of transmission of malaria, with an enduring threat of asymmetric military action and with a proactive approach by the chain of command to the implementation of malaria protection policy, no malaria cases developed. CONCLUSION: The chain of command can have a significant impact on compliance with malaria protection measures, which might reduce incidence of the disease in the deployed population. PMID- 15828693 TI - Cost analysis of military eye injuries in fiscal years 1988-1998. AB - Army, Navy, and Air Force safety center ground eye injuries were evaluated from fiscal year 1988 through fiscal year 1998. The U.S. Army Safety Center data revealed that the average Army military eye injury mishap caused a loss of 6.1 days from work, at a cost averaging 9724 dollars. The U.S. Navy Safety Center data showed that the average Navy military eye injury mishap caused a loss of 5.9 days from work, at a cost averaging 4222 dollars. The Air Force Safety Center data revealed that the average Air Force eye injury mishap caused a loss of 4.7 days from work, at a cost averaging 3196 dollars. Comparing safety center data with Defense Medical Surveillance System data exposed an underreporting of safety center data by at least 250%. To improve accurate meaningful data collection, an automated eye injury collection system should be initiated. PMID- 15828694 TI - Temporal lobectomy for refractory epilepsy in the U.S. military. AB - OBJECTIVES: To determine the characteristics, seizure outcomes, and quality-of life outcomes for military beneficiaries undergoing partial temporal lobectomy for refractory epilepsy at the only U.S. military medical center with a comprehensive epilepsy surgery program. METHODS: The records of all 84 patients treated with partial temporal lobectomy between 1986 and 2000 at Walter Reed Army Medical Center were retrospectively reviewed. Outcome measures included seizure frequency according to the Engel classification system, driving, employment, anticonvulsant use, and military service. RESULTS: The study cohort consisted of 72 military dependents, 10 active duty military members, and 2 military retirees. Two years after surgery, 65 (92%) of 71 patients had seizure improvement (Engel classes I-III) and 46 (66%) of 71 had seizure remission (Engel class I). Driving and employment rates increased after surgery, whereas anticonvulsant use decreased. Five (50%) of 10 active duty patients achieved seizure remission postoperatively and continued to serve in the Armed Forces. Active duty patients had a later age of seizure onset, shorter duration of epilepsy, and greater proportion of lesional epilepsy, compared with nonactive duty patients. CONCLUSIONS: Epilepsy surgery outcomes in the U.S. military are similar to those reported from nonmilitary centers, with the majority of patients experiencing seizure remission and improvements in quality-of-life measures. Complete seizure remission after successful anterior temporal lobectomy enables some active duty military members to continue service in the U.S. Armed Forces. PMID- 15828695 TI - Computed tomography of craniofacial trauma at a combat support hospital in Afghanistan. AB - Complex craniofacial injuries are encountered among both soldiers and civilians in combat zones. Computed tomography is a necessary and effective tool for the evaluation and treatment of these injuries in the forward-deployed combat support hospital. PMID- 15828696 TI - Delay in the diagnosis of Turkish servicemen with pulmonary tuberculosis. AB - OBJECTIVE: This study was designed to assess the time between the first appearance of symptoms and the start of treatment among Turkish servicemen with tuberculosis (TB). METHODS: Ninety-seven patients with pulmonary TB were enrolled in the study. We surveyed their complaints and education levels, the opportunity to visit a health care facility, and the time elapsed until diagnosis and treatment. RESULTS: Of the study group, 62 (63.7%) reported that they had visited an infirmary as soon as their complaints appeared. The median total delay in diagnosis was 21 days for all cases. Patient delay decreased among patients with hemoptysis (2.1 vs. 6.4 days, p = 0.013) and increased with night sweats (7.3 vs. 3.1 days, p = 0.042). Total delay was not correlated with any symptom. CONCLUSION: We suggest that delays in diagnosis and treatment among Turkish soldiers with pulmonary TB arise from some factors related to both patients and health care facilities, and these factors should be taken into account by military health services. PMID- 15828697 TI - An internet-based referral/consultation system for the U.S.-associated Pacific Islands: its contribution to orthopedic graduate medical education at Tripler Army Medical Center. AB - To evaluate the benefit of the Pacific Island Health Care Project (PIHCP) to our orthopedic graduate medical education program, we performed a retrospective review of our 8-year operative experience with patients referred through this program. Between July 1994 and June 2002, 69 patients underwent 79 orthopedic operative procedures. Patients were categorized by primary diagnosis, anatomic site involved, and surgical treatment rendered. Because many of the patients referred from the PIHCP with tumors were noted to have either unusually large lesions or advanced-stage disease, further analyses of tumor stage and pathologic grade were made. Seven of the 14 oncologic cases surgically treated in our department in the past 8 years were referrals from the PIHCP. Unique operative procedures performed for these tumor patients included one forequarter amputation, one hip disarticulation, one hemipelvectomy, two partial scapulectomies, and one distal ulna excision. We conclude that the PIHCP referrals provide an important and relatively unique contribution to the clinical and operative experience of our orthopedic residents. These patients from the Pacific basin also enhance our orthopedic graduate medical education program by exposing our residents to the special socioeconomic and cultural issues related to caring for people from developing insular countries. PMID- 15828698 TI - Predictors of length of stay in a psychiatric ward serving active duty military and civilian patients. AB - Admission data from 6366 patients on the psychiatry service at Naval Medical Center San Diego were used to form a linear regression model to examine variables that might influence length of stay. Information was available on active duty status, primary diagnosis, age, gender, and marital status. Active duty service, older age, single marital status, and a primary diagnosis of a psychotic or mood disorder were all significantly associated with longer hospital stays. Primary diagnosis of an adjustment or personality disorder was associated with a shorter stay. Taking into account these variables, lengths of stay for active duty personnel averaged 4.00 +/- 0.39 days longer than for equivalent civilian admissions. Although military personnel are generally thought of as a healthy population, psychiatric hospital stays were found to be longer in service members than in their civilian counterparts. Financial repercussions and possible reasons for this are discussed. PMID- 15828699 TI - Prevalence of psychotic comorbidity in combat-related post-traumatic stress disorder. AB - OBJECTIVE: We explored the prevalence of comorbidity of psychotic symptoms among Croatian war veterans with combat-related post-traumatic stress disorder (PTSD) and the psychotic features among these patients, as well as the association between psychotic symptoms and personality disorders. METHODS: The study included 680 men who had experienced combat stress and had diagnoses of PTSD confirmed before examination. They were compared with a group of 289 soldiers with combat experience under regular medical examination, who were still in active military service. The groups were matched in duration of combat experience, time elapsed between combat experience and the study, age, marital status, and education. Statistical analyses were performed using the t test and chi2 test. RESULTS: The study showed that 15% of war veterans had current chronic PTSD and 45% had PTSD with one or more comorbid diagnoses. Psychotic disorders with PTSD were confirmed for 17% of those patients and major depressive disorder with psychotic features for 15%. Depression and psychotic symptoms were more often found among patients without personality disorders. Also, we found that 9% of the war veterans who were still in active military service had some psychiatric disorders. CONCLUSIONS: Many patients demonstrated psychotic symptoms different from flashbacks and dissociative symptoms. Those psychotic symptoms are an integral part of PTSD and have a symbolic relation to the trauma. PMID- 15828700 TI - Building strong and ready Army families: a multirisk reduction health promotion pilot study. AB - A quasi-experimental prospective study was conducted to assess the stage of change progression and elimination of unhealthy behaviors among active duty Army soldiers and their spouses (N = 245) before and after attending a community-based wellness program. Significant differences in stage progression between the intervention group and a comparison group was seen in the fitness (p = 0.05) and communication risk categories (p = 0.03) immediately after the intervention; significant differences in stage progression in fitness (p = 0.05) and spirituality (p = 0.005) risk categories 6 months after baseline. There was a significant increase in seat belt use (p = 0.008), a decrease in reported tobacco use (p < 0.05), and a reduction in family stress (p = 0.06) and personal stress (p < 0.02) in the intervention group. Targeting multiple behavior changes and risk reduction in a single intervention program presents several challenges. In this pilot study, there were many positive nonsignificant trends in risk reduction that might translate into significant changes with a larger sample. PMID- 15828701 TI - Declining access to alcohol and drug abuse services among veterans in the general population. AB - Although the Veterans Health Administration has been committed to preserving its capacity to provide specialized substance abuse (SA) services, administrative data from the late 1990s point to reduced access. To explore the possible effects of reduced access to Veterans Affairs services on veterans in the general population, we examined data from the National Household Survey of Drug Abuse from 1994 to 2001. Although the data are not longitudinal at the level of individuals, the annual nature of these surveys makes it possible to chart aggregate changes in clinical needs and service use over time. The results indicate that, whereas veterans' needs for SA services remained high and were consistent across these years, the proportion of veterans in treatment decreased. The proportion of nonveterans receiving SA treatment also decreased but not as steeply as that of veterans. This trend parallels declining delivery of specialized alcohol and drug abuse services within the Veterans Health Administration, although they do not prove a causal relationship. PMID- 15828702 TI - The use of World War II chest radiograph in the identification of a missing-in action U.S. Marine. AB - A World War II mass grave was recovered in 1999 by a U.S. Army team and yielded 20 complete skeletons. A case study involving the identification of one of these individuals is presented in this article. The thought processes and problems that presented themselves to the forensic anthropologist and odontologist are detailed. Methods used to establish identity are described. This case demonstrates how standard operating procedures used by a forensic anthropologist and odontologist can narrow the field of possible individuals associated with remains, and with extra information--in this case, a military radiograph taken in 1941--can ultimately establish the identity of a decedent. The authors learned that some medical records, which at first glance appear to be excess or irrelevant, may contain the item required to be certain that a case is strong in support of a recommended identification. PMID- 15828703 TI - Do clinical practice guidelines improve processes or outcomes in primary care? AB - BACKGROUND: Clinical practice guidelines (CPGs) are common, but it is not clear whether they improve care. METHODS: Quality indicators for processes and outcomes of care were obtained from a computerized system-wide database by patient administration and utilization management personnel unaware of this study and without connection to or interests in guideline implementation. These indicators were compared before and after guideline implementation. RESULTS: After the asthma CPG, nebulizer treatments, emergency department visits, and admissions decreased significantly (p < 0.001 for all three) and education increased significantly (p < 0.001). Periodic measurements of lung function and controller medication prescriptions were unchanged. After the diabetes mellitus CPG, microalbumin screens and education increased significantly (p < 0.001). Angiotensin-converting enzyme inhibitor prescriptions and yearly foot examinations decreased significantly, along with the percentage of patients with blood pressure of < or = 130/85 mm Hg (p < 0.001). Mean hemoglobin A(1C) levels did not change significantly. After the tobacco cessation CPG, screening and education increased significantly (p < 0.001 and p = 0.04, respectively). CONCLUSIONS: The asthma CPG improved some processes and all outcomes. The diabetes CPG improved two of the eight measured processes but had no effect on outcomes. Education and screening, but not counseling, improved with the tobacco CPG. CPGs appear to improve diagnostic and educational processes more than provider-dependent treatment processes. Outcomes were improved after implementation of the asthma CPG but not after the diabetes CPG. PMID- 15828704 TI - An evaluation of the leadership attitudes of managers in Turkish Armed Forces' hospitals. AB - BACKGROUND: As in any other institution, the success of hospitals, which use approximately one-half of all resources in the health sector, depends on moving human resources toward the institution's goals. Ensuring that the workers are used intentionally for specific, predetermined goals, and that the workers are motivated toward achieving these institutional goals are the responsibilities of the managers. OBJECTIVE: In this study, the leadership attitudes of the higher rank executive officers in Turkish Armed Forces' military hospitals were evaluated. METHODS: The managerial grid developed by Blake and Mouton was used to evaluate the leadership attitudes of the officers. A total of 172 questionnaires was sent out to managers, but only 142 (82.6%) were returned completed. RESULTS: The study results show that the managers substantially preferred team leadership styles in terms of leadership philosophy, planning, and evaluation functions, but preferred task-oriented leadership and mid-way leadership styles in terms of execution. Also, it was found that 72% of all managers practiced team leadership in terms of the overall leadership manner. CONCLUSIONS: It seems as if the type of leadership participants preferred was team leadership, and the least was passive leadership. From this finding, we suggest that the managers in Turkish Armed Forces' hospitals are not only interested in their work and goals, but also in the interests of their workers as human beings. PMID- 15828705 TI - Cadaver testing to validate design criteria of an adult intraosseous infusion system. AB - INTRODUCTION: The FAST 1 intraosseous (IO) infusion system was designed to deliver fluids and medications into the adult sternum in the prehospital and battlefield environments. OBJECTIVE: To test the prototype in 106 cadavers and excised sterna and compare it with other IO devices. RESULTS: The insertion force was similar to that of other IO devices (mean, 8.5 kg; range, 2.3-19.6 kg). In 39 of 39 trials, the depth-control mechanism inserted the portal within 1.0 mm of a predetermined distance below the anterior surface of the cortical bone. If misplaced, underpenetration was more likely than overpenetration (mean displacement, -0.3 mm; SD, 0.5 mm). After release, the portal could not be advanced further into the manubrium. Marrow was accessed in 75 of 77 trials. Mean flow rates were 109 mL/min for normal saline solution and 102 mL/min for hypertonic saline/dextran, similar to the Cook Sur-Fast device. CONCLUSION: The cadaver and bench tests demonstrated the reliability and safety of the FAST 1 system at the design/prototype stage. PMID- 15828706 TI - Form and function in craniofacial deformities. AB - The interplay of form and function is recognized throughout nature. Whether at the cellular level or visible form, physiological function will not be optimal if not supported by ideal morphology. This principle could not be more true than in the relationship between the human skull and face. The development of ideal skull and facial skeletal form is critical for the function of the brain, vision, airway, mastication, and speech. When craniofacial structure is altered by birth defects, proper functioning is drastically affected. We review the neurocranial basis for normal craniofacial skeletal development and present craniofacial abnormalities that illustrate their deleterious affect on facial function. PMID- 15828707 TI - The circle of sagittal synostosis surgery. AB - Sagittal synostosis, the premature closure of the sagittal suture, accounts for more than 50% of all nonsyndromic single-suture synostoses. Although no detrimental neurologic effects can be directly attributed to the synostosis, a number of patients will have relatively increased intracranial pressure. Surgical correction for sagittal synostosis has evolved from simple removal of bone strips to extensive cranial remodeling, all in a attempt to achieve a normal head shape. The lack of outcome measures has limited the surgeon's ability to choose one surgical procedure over another. The use of a cranial helmet for passive molding of the head after surgery is effective when used with limited endoscopic surgery. We present the results of using an extended strip craniectomy combined with long duration molding helmet therapy and compare it with other reported methods. The results suggest that passive molding with the helmet may have a greater effect on cranial shape than surgery. PMID- 15828708 TI - Brief synopsis of cranial sutures: optimization by adaptation. AB - This article reviews the form and function of cranial sutures across the temporal and spatial scales. The temporal scale spans 530 million years, from ostracoderms to contemporary humans. The spatial scale spans eight orders of magnitude, from the macroarchitectural level (the entire cranium), through the mesoarchitectural (the local/regional bone-suture-bone complex) and microarchitectural levels (tissues and cells), to the nanoarchitectural level (molecules within and outside the cells). A mechanomorphologic loop, or cycle, exists. The mechanical strain experienced by the sutures eventually alters the morphology of the sutures. In turn, these morphological changes affect the strain distribution within and around the sutures. At the microarchitectural level, the responses of bone and sutural cells to environmental perturbations depend on the content (what that perturbation is), the context (the other coexisting extrinsic and intrinsic factors), and the history of the perturbation (how often and for how long). PMID- 15828709 TI - Craniosynostosis syndromes in the genomic era. AB - The origin of craniosynostosis is heterogeneous: hereditary, mechanical, teratogenic, and idiopathic. Craniosynostosis is further defined by the suture(s) involved and whether it is syndromic or nonsyndromic. Syndromic craniosynostosis typically involves cranial sutures plus central nervous system and extracranial skeletal changes. Nonsyndromic craniosynostosis is usually confined to cranial changes. The most common syndromic synostoses reflect changes in fibroblast growth factor receptor (FGFR) activity related to mutations in the genes coding for these receptors. Other genes have been implicated in craniosynostosis syndromes. Several craniosynostosis syndromes are caused by mutation of the same FGFR, making the eponymic designation (eg, Crouzon's or Pfeiffer's syndrome) unclear. Ultimately, syndrome eponyms may be replaced by designation of the underlying mutation. Neurologic complications may include mental retardation, increased intracranial pressure, and cranial nerve abnormalities. Craniosynostosis syndromes require careful physical examination, radiological investigation, and now molecular evaluation to predict outcome and risk of recurrence. PMID- 15828710 TI - The effects of craniosynostosis on the brain with respect to intracranial pressure. AB - Intracranial pressure (ICP) and skull volume are intricately related. Craniosynostosis alters skull volume, and the many forms of craniosynostosis complicate the relationship to ICP even further. Patients with single-suture synostosis are less likely to experience elevated ICP than patients in whom multiple sutures, craniofacial syndromes, or both are involved. Among patients with more than one suture involved, the multifactorial mechanisms underlying elevated ICP include cephalocranial disproportion and venous outflow obstruction. Direct monitoring of ICP for at least 24 hours can aid in the diagnosis and decision making process. The management of craniosynostotic patients is diverse and necessitates a long-term plan for follow-up. PMID- 15828711 TI - Cranial remodeling devices: treatment of deformational plagiocephaly and postsurgical applications. AB - Since the first cranial remodeling devices were introduced in 1979, both their design and availability have continued to evolve. Today, these devices are used to treat deformational plagiocephaly (plagiocephaly, brachycephaly, and dolichocephaly) and are used as adjuncts to surgery for craniosynostosis. In deformational plagiocephaly, the goal is to improve cranial symmetry and return the cranium to a more normal proportion. Postoperatively, these devices are used to provide stabilization and to enhance surgical outcomes. Numerous clinical studies have demonstrated the safety and efficacy of these devices by documenting statistically significant reductions in the cranial vault, skull base, and facial asymmetries as well as improvements in the cephalic index. These studies indicate that cranial remodeling devices play an important role in the treatment of cranial deformations. PMID- 15828712 TI - Blood sparing in craniosynostosis surgery. AB - One of the main risks of craniosynostosis surgery is the possible need for an allogenic blood transfusion (ABT). Most patients are operated on in the first months of life, when physiological conditions are particularly sensitive to even limited blood losses. Furthermore, most surgical techniques proposed in the past were based on extensive craniectomies and cranial remodeling. Because of the known infective and immunologic risks of ABT, in recent years more attention has been dedicated to factors that might help reduce the risk of ABT. We review recent preoperative (ie, erythropoietin administration), intraoperative (ie, acute normovolemic hemodilution, intraoperative blood salvage), and postoperative (ie, clinical monitoring, postoperative blood salvage) anesthesiologic procedures developed with this aim in mind. We also consider operative techniques and technical apparatus that reduce surgical invasiveness, particularly preoperative planning, age selection, and the role of endoscopic assistance and gradual distraction devices. PMID- 15828713 TI - Craniofacial surgery: complications and their prevention. AB - This article focuses on the management of perioperative complications in surgery dealing with structures of the craniofacial complex. A brief review of applicable surgical techniques is given, beginning with single-suture craniosynostoses and moving on to the more complex syndromic craniofacial disorders. Distinctive complications associated with each technique are discussed. PMID- 15828714 TI - On the current incidence of deformational plagiocephaly: an estimation based on prospective registration at a single center. AB - In 1992, the American Academy of Pediatrics (AAP) recommended supine sleeping to reduce the risk of sudden infant death syndrome. Although the incidence of deformational plagiocephaly is unknown, the consensus is that it has increased since this recommendation was made. To estimate the current incidence of plagiocephaly, we examined 342 infants for signs of deformational plagiocephaly, including occipital flattening, ear misalignment, frontal bossing, and facial asymmetry. Noticeable occipital flattening was documented in 15.2% of the infants (95% confidence interval, 11.6% to 19.5%); 1.46% had significant cranial deformities that also affected the skull base and face. Significant cranial asymmetry, defined as occipital flattening with concomitant skull base involvement and facial asymmetry, was observed in almost 1 in 68 infants. Adding to a growing body of evidence, our findings suggest significant increases in clinical deformational plagiocephaly since initiation of the AAP's "Back to Sleep" campaign. PMID- 15828715 TI - Cognitive impact of craniosynostosis. AB - This article reviews the data on the effects of synostosis on cognitive development and the role of surgical intervention in ameliorating these effects. The literature on children with single-suture craniosynostosis, treated both surgically and conservatively, is reviewed. The evidence for possible pathophysiological mechanisms by which brain restriction might affect cognitive development is also reviewed. Although children with single-suture isolated craniosynostosis may be at risk of developmental delay, learning disability, or both, especially with regard to speech or language skills, available testing methodologies provide no evidence of an association between surgical intervention and ultimate intellectual outcome. Despite the controversies about the role of surgery in addressing the cognitive sequelae of simple craniosynostosis, in the absence of definitive data, the indications for surgery include correction of deformity and minimization of cognitive sequelae. The relative risks of performing surgery early or late must be determined on an individual basis, balancing potential cosmetic and cognitive benefits against the heightened risk of reoperation or perioperative morbidity. PMID- 15828716 TI - Anything to declare? PMID- 15828717 TI - Changes to the Compulsory Scrapie Flocks Scheme. PMID- 15828718 TI - Revised contingency plan on exotic disease outbreaks. PMID- 15828719 TI - RCVS specialist in animal welfare science, ethics and law. PMID- 15828721 TI - Brucellosis suspected in Derbyshire. PMID- 15828720 TI - New application form for project licences. PMID- 15828722 TI - Salmonella Dublin in cattle in Scotland: fall in the number of outbreaks continues. PMID- 15828723 TI - Frequencies of PrP genotypes in 38 breeds of sheep sampled in the National Scrapie Plan for Great Britain. AB - Between October 2001 and January 2003 the prion protein (PrP) genotypes of over 250,000 sheep were determined through the operation of the National Scrapie Plan (NSP); the results for 38 breeds were analysed to provide an estimate of the underlying PrP genotype distribution of the British sheep population. Although there was marked variability among the genotype profiles of the different breeds, several trends emerged. A comparison of the allele frequencies demonstrated that the breeds could be grouped into three categories: breeds dominated by ARR and ARQ in which the frequency of ARR exceeded the frequency of ARQ; breeds dominated by ARR and ARQ in which the frequency of ARQ exceeded the frequency of ARR; and breeds with significant levels of either AHQ, ARH or VRQ. Hill breeds were more likely to have a lower proportion of animals at low risk of scrapie (NSP type 1) and a higher proportion of animals at an intermediate risk of scrapie (NSP type 3) than other breeds. Most breeds had a small proportion of animals at high risk of scrapie (NSP type 5). The frequency of ARR/VRQ (NSP type 4) was variable. PMID- 15828724 TI - Clinical findings and treatment of 30 cattle with botulism. AB - The clinical signs, the results of haematological and biochemical analyses and the treatment of 30 cattle with botulism are described, and the signs of the 13 cattle that survived are compared with those of the 17 that were euthanased owing to the disease. The cattle originated from 11 farms that had experienced an outbreak of botulism. The most important clinical sign in all the cattle was a reduction in the strength of the tongue; excessive salivation and difficulty in swallowing were observed in 20 of the animals, and the ears of 15 of them drooped. In 21 of the cattle, reaction to pricking of the head and body with a hypodermic needle was either absent or slight. Twelve of the animals had an unsteady, slow, difficult gait, and nine were unable to stand. A significantly higher proportion of the cattle which were euthanased had marked changes in behaviour and condition, anorexia, severely reduced skin turgor, weak tongues, a low rectal temperature, a high heart rate and a low blood pH; 11 were euthanased immediately after a clinical examination and six were euthanased one to five days after the initiation of treatment because their condition had deteriorated. Thirteen of the animals were treated for three to 23 days and were healthy when they were discharged. The treatment consisted of an intravenous infusion of 10 to 20 litres of glucose saline per day and the daily administration of fresh ruminal juice. Follow-up by telephone several months later revealed that all 13 animals had recovered completely. PMID- 15828725 TI - Myocarditis and myositis due to infection with Hepatozoon species in pine martens (Martes martes) in Scotland. AB - Postmortem examinations of four pine martens which had died as a result of road accidents in Scotland revealed focal, granulomatous lesions in the heart and skeletal muscles of three of them. An immunoperoxidase staining technique showed that the lesions were due to infection with Hepatozoon species. A PCR-based assay was used to confirm the presence of Hepatozoon DNA in the infected tissues. The nucleotide base sequence of the PCR products suggested that the infecting organism was probably a new species of Hepatozoon, most closely related to, but distinct from, Hepatozoon canis. The pine martens were in good physical condition and there was no indication that the infection was causing ill health. PMID- 15828726 TI - Evaluation of probiotics as a substitute for antibiotics in a large pig nursery. PMID- 15828727 TI - Persistent efficacy of doramectin against Haemonchus contortus in goats. PMID- 15828728 TI - First autochthonous case of canine ocular onchocercosis in Germany. PMID- 15828729 TI - Huevil (Vestia foetida) poisoning of cattle in Chile. PMID- 15828730 TI - Corneal papilloma in a dog. PMID- 15828731 TI - Teeth, bones and brushing. PMID- 15828732 TI - Bovine TB and badgers. PMID- 15828733 TI - Equine polysaccharide storage myopathy. PMID- 15828734 TI - Treating pet poultry. PMID- 15828742 TI - Study of 253 dogs in the United Kingdom with diabetes mellitus. AB - Clinical information and blood samples were collected from 253 dogs with naturally occurring diabetes mellitus. Over half of them were labrador retrievers, collies, Yorkshire terriers or crossbred dogs, and approximately 80 per cent of them were diagnosed between the ages of five and 12 years. The majority of the dogs were receiving insulin therapy once a day, but in the dogs receiving insulin injections twice a day there was a trend for lower serum fructosamine concentrations, suggesting better glycaemic control. The proportion of female dogs with diabetes was lower than in previous surveys. The disease was diagnosed more commonly in the winter months, a seasonal pattern also observed in human beings with diabetes, suggesting that similar environmental factors might be involved in the disease. PMID- 15828743 TI - Neuropathological findings in cats with clinically suspect but histologically unconfirmed feline spongiform encephalopathy. AB - Central nervous system (CNS) tissues from 192 cats with neurological signs were examined histologically, and tissues from 173 of them were later examined immunohistochemically as part of a survey to determine the prevalence of feline spongiform encephalopathy (FSE). One of the cats was from Norway and the others were from Great Britain. The most commonly recorded clinical signs were ataxia, behavioural changes and epilepsy, but none of the cats had histopathological evidence of FSE. The most common organic CNS lesions were non-suppurative encephalomyelitis in 28 per cent, neoplasia in 15 per cent and a heterogeneous group of degenerative encephalopathies in 9 per cent of the cats. A range of minor histological lesions of uncertain significance was also observed. No histological lesions were observed in the tissues of 63 (33 per cent) of the cats. Disease-specific prion protein (PrP(Sc)) was observed in only one of the 173 cats examined by immunohistochemistry. PMID- 15828744 TI - Kinematic analysis of the gait of 10 labrador retrievers during treadmill locomotion. AB - The trotting gait of 10 sound, adult labrador retrievers was analysed using kinematic gait analysis on a purpose-built treadmill using video-based motion analysis software. The maximal angular displacement, minimal angular displacement, average angular displacement, and the maximal positive and negative angular velocities of the right elbow and right stifle were measured over five gait cycles at defined time points during each of five two-minute sessions. The dogs' trotting gait was not repeatable, either for individual dogs during the first session or between sessions, or between dogs at the same time points during a session. PMID- 15828745 TI - Intranasal kennel cough vaccine protecting dogs from experimental Bordetella bronchiseptica challenge within 72 hours. PMID- 15828746 TI - Molecular diagnosis of feline sporotrichosis. PMID- 15828747 TI - Evaluation of the Schirmer and phenol red thread tests for measuring tear secretion in rabbits. PMID- 15828748 TI - Pieris japonica intoxication in an African spurred tortoise (Geochelone sulcata). PMID- 15828749 TI - Prescribing costs. PMID- 15828750 TI - Bovine TB and badgers. PMID- 15828751 TI - Teeth, bones and brushing. PMID- 15828752 TI - Regulation of paraprofessionals. PMID- 15828753 TI - Treating pet poultry. PMID- 15828755 TI - Veterinary Computer Users group. PMID- 15828756 TI - Teeth, bones and brushing. PMID- 15828757 TI - Teeth, bones and brushing. PMID- 15828758 TI - Selective determination of the doxorubicin content of individual acidic organelles in impure subcellular fractions. AB - Since organelle preparations often contain more than one organelle type (e.g., acidic organelles and mitochondria), techniques that measure the properties of a given organelle type while avoiding biases caused by ancillary subcellular compartments are highly desirable. We report here the use of capillary electrophoresis (CE) with laser-induced fluorescence (LIF) dual-channel detection to identify acidic organelles containing doxorubicin (DOX) in crude subcellular fractions from CCRF-CEM and CEM/C2 cell lines. As confirmed by confocal microscopy, acidic organelles are identified by their accumulation of fluorescently labeled nanospheres. Using CE-LIF analysis, individually detected organelles are classified into three kinds: acidic organelles containing only nanospheres, acidic organelles containing nanospheres and DOX, and other organelles containing DOX (e.g., mitochondria) with no detectable nanospheres. Electrophoretic mobility, DOX fluorescence intensity, and nanosphere fluorescence intensity distributions of individual acidic organelles and other organelles containing DOX are determined in the same CE-LIF run. The acidic organelle mobilities range from (-0.7 to -2.0) x 10(-4) cm(2) V(-1) s(-1) while those of the other organelles spread from (-0.6 to -3.5) x 10(-4) cm(2) V(-1) s(-1). In addition, by calibrating the detector response, DOX content in individual acidic organelles and other organelles can be estimated. The average amounts of DOX per acidic organelle in CEM/C2 and CCRF-CEM cells are 11.1 +/- 0.5 and 10.6 +/- 0.4 zmol, respectively. This first report of an analysis of the accumulation of DOX in individual acidic organelles presents a procedure for analyzing the accumulation of fluorescent compounds in acidic organelles that could be useful for investigating acidic organelle maturation and the role of these organelles in drug resistance. PMID- 15828759 TI - Application of time-of-flight mass spectrometry with laser-based photoionization methods for time-resolved on-line analysis of mainstream cigarette smoke. AB - The application of soft photoionization mass spectrometry methods (PIMS) for cigarette mainstream smoke analysis is demonstrated. Resonance-enhanced multiphoton ionization (REMPI) at 260 nm and vacuum ultraviolet light single photon ionization (SPI) at 118 nm were used in combination with time-of-flight mass spectrometry (TOFMS). An optimized smoking machine with reduced memory effects of smoke components was constructed, which in combination with the REMPI/SPI-TOFMS instrument allows PIMS smoke analysis with a time resolution of up to 10 Hz. The complementary character of both PIMS methods is demonstrated. SPI allows the detection of various aliphatic and aromatic compounds in smoke up to approximately 120 m/z while REMPI is well suited for aromatic compounds. The capability of the instrument coupled to the novel sampling system for puff-by puff resolved measurements is demonstrated. The feasibility of using the experimental system for intrapuff smoke measurements is also shown. Two main patterns of puff-by-puff behaviors are observed for different smoke constituents. The first group exhibits a constant increase in smoke constituent yield from the first to the last puff. The second group shows a high yield of the constituent in the first puff, with lower and constant or slowly increasing yields in the following puffs. A third group cannot be clearly classified and is a combination of both observed profiles. PMID- 15828760 TI - Versatile new ion source for the analysis of materials in open air under ambient conditions. AB - A new ion source has been developed for rapid, noncontact analysis of materials at ambient pressure and at ground potential. The new source, termed DART (for "Direct Analysis in Real Time"), is based on the reactions of electronic or vibronic excited-state species with reagent molecules and polar or nonpolar analytes. DART has been installed on a high-resolution time-of-flight mass spectrometer (TOFMS) that provides improved selectivity and accurate elemental composition assignment through exact mass measurements. Although DART has been applied to the analysis of gases, liquids, and solids, a unique application is the direct detection of chemicals on surfaces without requiring sample preparation, such as wiping or solvent extraction. DART has demonstrated success in sampling hundreds of chemicals, including chemical agents and their signatures, pharmaceutics, metabolites, peptides and oligosaccharides, synthetic organics, organometallics, drugs of abuse, explosives, and toxic industrial chemicals. These species were detected on various surfaces, such as concrete, asphalt, human skin, currency, airline boarding passes, business cards, fruits, vegetables, spices, beverages, body fluids, horticultural leaves, cocktail glasses, and clothing. DART employs no radioactive components and is more versatile than devices using radioisotope-based ionization. Because its response is instantaneous, DART provides real-time information, a critical requirement for screening or high throughput. PMID- 15828761 TI - Probing strong adsorption of solute onto C18-silica gel by fluorescence correlation imaging and single-molecule spectroscopy under RPLC conditions. AB - Understanding molecular adsorption at a chromatographic interface is of great interest for addressing the tailing problem in chemical separations. Single molecule spectroscopy and confocal fluorescence correlation imaging are used to study the adsorption sites of C(18) silica beads under RPLC chromatographic conditions. The experiments show that cationic molecule rhodamine 6G laterally diffuses through the chromatographic interface of a C(18) hydrocarbon monolayer and acetonitrile with occasional reversible strong adsorptions. Fluorescence correlation imaging extracts the rare strong adsorption events from large data sets, revealing that the strong adsorption sites are randomly distributed throughout the silica beads. Virtually every imaging pixel of silica beads adsorbs molecules. Single-molecule spectroscopy of the 584 strong adsorption events observed indicates that the strong adsorptions persist on the time scales from several milliseconds to seconds, having an average desorption time of 61 ms. The strong adsorption events are rare, comprising 0.3% of the total observation time. The sizes of strong adsorption sites are within the optical resolution of confocal imaging. PMID- 15828762 TI - Multiple and simultaneous detection of specific bacteria in enriched bacterial communities using a DNA microarray chip with randomly generated genomic DNA probes. AB - A DNA microarray chip for detecting the presence of specific bacterial strains was developed using random genomic probes derived from genomic DNA, i.e., without any sequence information. Thirteen bacteria from different genuses were selected as targets. For the fabrication of the random genomic probes, genomic DNA from pure cultures of each bacterium was fractionated using several pairs of restriction endonucleases. After size fractionation of the genomic DNA fragments, random genomic libraries for each bacterium were constructed. From the library, specific probes were amplified by PCR and the probes were affixed to a slide glass to fabricate the DNA microarray chip. The results from tests with pure and mixed cultures of the bacteria used in the fabrication of the chips showed specific responses and only a small portion of cross-hybridization. This DNA microarray chip was also tested to detect the presence of specific bacteria in mixed populations. In these tests, it was demonstrated that this system provided a fast and specific response to the presence of bacterial species in mixed samples, even in activated sludge samples. This indicates that any DNA microarray chip for the detection of specific bacteria can be fabricated using the same protocols as presented in this study without requiring any genus level sequence information from pure isolates. PMID- 15828763 TI - Genetically engineered elastin-protein A fusion as a universal platform for homogeneous, phase-separation immunoassay. AB - A simple and universal platform for competitive phase-separation immunoassay is reported based on a fusion protein composed of a temperature-responsive elastin like polypeptide (ELP) and the antibody-binding staphylococcal protein A (SpA). The basic principle is to take advantage of the ability of SpA to bind a variety of antibodies with high affinity, allowing simple separation of antigen-antibody complex by thermal precipitation. The resulting ELP-SpA fusion was shown to preserve the ability to reversibly precipitate as well as its high affinity toward different IgGs and IgMs. As a model system, a competitive phase-separation immunoassay based on the ELP-SpA format was established for paclitaxel (taxol) with IC(50) (20.18 nM) and the lower detection limit (2.94 nM) very similar to those reported for the ELISA format. Unlike the heterogeneous interaction in ELISA, which decreases the antibody-binding activity, the reported homogeneous immunoassay not only alleviates this problem but also enables the potential for high-throughput automation. We believe that the reported ELP-SpA fusion will find applications not only as a powerful diagnostic tool for diverse analytes but also a potential useful tool for purification and immobilization of antibody. PMID- 15828764 TI - High-speed, high-resolution monolithic capillary LC-MALDI MS using an off-line continuous deposition interface for proteomic analysis. AB - High-speed, high-resolution LC separations, using a poly(styrene-divinylbenzene) monolithic column, have been coupled to MALDI MS and MS/MS through an off-line continuous deposition interface. The LC eluent was mixed with alpha-cyano-4 hydroxycinnamic acid matrix solution and deposited on a MALDI plate that had been precoated with nitrocellulose. Deposition at subatmospheric pressure (80 Torr) formed a 250-microm-wide serpentine trace with uniform width and microcrystalline morphology. The deposited trace was then analyzed in the MS mode using a MALDI TOF/TOF MS instrument. Continuous deposition allowed interrogation of the separation with a high data sampling rate in the chromatographic dimensions, thus preserving the high resolution of narrow peaks (3-5-s peak width at half-height) of the fast monolithic LC. No extracolumn band broadening due to the deposition process was observed. Over 2000 components were resolved in a 10-min linear gradient separation of the model sample, and 386 unique peptides were identified in the subsequent MS/MS analysis. The continuous deposition interface allows the coupling of high-resolution separations to MALDI MS without degradation in separation efficiency, thus enabling high-throughput proteome analysis. PMID- 15828765 TI - Increasing the efficiency of in-capillary electrophoretically mediated microanalysis reactions via rapid polarity switching. AB - We report herein a new approach to enhance the sensitivity or speed of CE-based methods that involve in-line reactions. Rapid polarity switching (RPS) is used as a novel means for in-line mixing of two reactant solutions via rapid (1-5 s) and sequential switching of the applied potential field. By employing the RPS approach with a model chemical reaction, that between creatinine and alkaline picrate, significant enhancement in sensitivity (or a decrease in analysis time) is realized. Both increased convection and electrophoretic stacking of the ionic reagent appear to contribute to the rise in apparent reaction rate. When coupled with in-line chemistry of the Jaffe method for creatinine, the RPS methodology allows for 3-fold faster determination of creatinine in the concentration range needed for the analysis of clinical blood serum specimens. The new approach also allows the analysis to be performed without the need for the cumbersome and problematic enhanced sensitivity cell. PMID- 15828766 TI - A general method for patterning gradients of biomolecules on surfaces using microfluidic networks. AB - This report outlines a general method for the fabrication of immobilized gradients of biomolecules on surfaces. This method utilizes a microfluidic network that generates a gradient of avidin in solution and immobilizes this protein on the surface of glass or poly(dimethylsiloxane) by physical adsorption. The immobilized gradient of avidin is then translated into gradients of biotinylated ligands (e.g., small molecules, oligomers of DNA, polysaccharides) using the specific interaction between biotin and avidin. This method can also generate immobilized gradients of certain proteins and artificial polymers by a direct transfer of gradients from solution onto the surface. The major advantage of this method is that almost any type of molecule can, in principle, be immobilized in a well-defined surface gradient of arbitrary shape with dimensions of a few micrometers to a few centimeters. It is possible to tailor the precise shapes of gradients on surfaces from gradients in solution, either kinetically or competitively. Kinetic methods rely on controlling the time that the surface is exposed to the gradient in solution: when a single protein adsorbs from solution, the amount that adsorbs depends both on its concentration in solution and on the time allowed for adsorption. Competitive methods rely on exposure of the surface to a complementary gradient of two proteins in solution (In these experiments, the sum of the concentrations of the proteins in solution is independent of positions although the concentration of each, individually, depends on the position. In this procedure, the relative amount of each protein, at saturation on the surface, depends only on its concentration.). PMID- 15828767 TI - Identification of major histocompatibility complex-regulated body odorants by statistical analysis of a comparative gas chromatography/mass spectrometry experiment. AB - This paper examines the application of gas chromatography/mass spectrometry (GC/MS) in a comparative experiment to identify volatile compounds from urine that differ in concentration between two groups of inbred mice. A complex mixture might comprise several hundred or even thousands of volatile compounds. Because their number and location in a chromatogram are generally unknown, and because components overlap in populous chromatograms, the statistical problems offer significant challenges beyond traditional two-group screening procedures. We describe a statistical procedure to compare two-dimensional GC/MS profiles between groups, which entails (1) signal processing, baseline correction, and peak detection in single ion chromatograms; (2) aligning chromatograms in time; (3) normalizing differences in overall signal intensities; and (4) detecting chromatographic regions that differ between groups. In an application to chemosignaling, we detect differences in GC/MS chromatograms of ether-extracted urine collected from two inbred groups of mice that differ only in genes of the major histocompatibility complex (MHC). Several dozen MHC-regulated compounds are found, including two known mouse pheromones, 2,5-dimethylpyrazine and 2-sec-butyl 4,5-dihydrothiazole. PMID- 15828768 TI - Affinity monoliths for ultrafast immunoextraction. AB - Affinity monoliths based on a copolymer of glycidyl methacrylate and ethylene dimethacrylate were developed for ultrafast immunoextractions. Rabbit immunoglobulin G (IgG) and anti-FITC antibodies were used as model ligands for this work. The antibody content of the monoliths was optimized by varying both the polymerization and immobilization conditions for preparing such supports. The temperature and porogen composition used during polymerization showed significant effects on monolith morphology and on the amount of antibodies that could be coupled to these materials. The effects of various immobilization procedures and coupling conditions were also evaluated, including the coupling temperature, pH, protein concentration, and use of high buffer concentrations. The maximum ligand density obtained for rabbit IgG was approximately 60 mg/g. When a 4.5 mm i.d. x 0.95 mm monolith disk containing anti-FITC antibodies was used, 95% extraction of fluorescein was achieved in 100 ms. These properties make such monoliths attractive for work in the rapid isolation of analytes from biological samples. Similar columns can be developed for other targets by varying the types of antibodies or binding agents placed within the monoliths. PMID- 15828769 TI - A deoxynucleotide derivatization methodology for improving LC-ESI-MS detection. AB - We have developed a novel LC-UV-MS derivatization method for the analysis of deoxyguanosine monophosphate adducts that demonstrates enhanced signal intensities relative to underivatized analytes in positive ion mode electrospray ionization MS. Detection of DNA nucleotide adducts is normally conducted in negative ion mode, which requires basic mobile phases that make chromatographic separations difficult and reduce MS sensitivity. Utilizing coupling reagents typically employed in peptide synthesis, several different deoxyguanosine nucleotide phosphoramidates and phosphomonoesters were synthesized in high conversion yield and under mild reaction conditions. The derivatives were characterized by MS/MS and reaction conversion yields determined from the DAD-UV traces. The derivatives were evaluated for ionization efficiencies, fragmentation patterns, and reversed-phase chromatographic properties by LC/ESI-MS/MS. Overall, the hydrophobic derivatives showed increases in ionization efficiency and improved peak shape. Rank ordering of the derivatizing agents was initially established using the dGp-modified derivatives. The best derivatizing agent, hexamethyleneimine, showed a 3-4-fold signal enhancement compared to underivatized dGp and was selected for additional evaluation. A model system using the carcinogen, N-acetoxy-2-acetylaminofluorene (AAAF), was used to synthesize a N-acetyl-(2-aminofluorenyl)-guanosine 5'-monophosphate (dGpAAF) adduct, which was subsequently derivatized with hexamethyleneimine. Detection limits for dGphex and dGpAAFhex, purified by HPLC, were 10- and 3-fold higher (S/N) than their respective underivatized analogues. Practical applicability, with similar improvements in sensitivity, was established by derivatizing adducts isolated from calf thymus DNA exposed to AAAF. Our results demonstrate the utility of simple reactions for the enhanced detection of a mononucleotide in positive ion mode ESI MS and the application of this technique for the detection of dGp-DNA adducts at the low-femtomole level. PMID- 15828770 TI - Optical probes for biological applications based on surface-enhanced Raman scattering from indocyanine green on gold nanoparticles. AB - We report surface-enhanced Raman scattering (SERS) studies on indocyanine green (ICG) on colloidal silver and gold and demonstrate a novel optical probe for applications in living cells. In addition to its own detection by the characteristic ICG SERS signatures, the ICG gold nanoprobe delivers spatially localized chemical information from its biological environment by employing SERS in the local optical fields of the gold nanoparticles. The probe offers the potential to increase the spectral specificity and selectivity of current chemical characterization approaches of living cells and biomaterials based on vibrational information. PMID- 15828771 TI - Affinity chromatographic selection of carbonylated proteins followed by identification of oxidation sites using tandem mass spectrometry. AB - It has been shown that oxidatively modified forms of proteins accumulate during oxidative stress, aging, and in some age-related diseases. One of the unique features of a wide variety of routes by which proteins are oxidized is the generation of carbonyl groups. This paper reports a method for the isolation of oxidized proteins, which involves (1) biotinylation of oxidized proteins with biotin hydrazide and (2) affinity enrichment using monomeric avidin affinity chromatography columns. The selectivity of the method was validated by adding in vitro oxidized biotinylated BSA to a yeast lysate and showing that the predominant protein recovered was BSA. This method was applied to the question of whether large doses of 2-nitropropane produce oxidized proteins. A study of rat liver homogenates showed that animals dosed with 2-nitropropane produced 17 times more oxidized protein than controls in 6 h. Tryptic digestion of these oxidized proteins followed by reversed-phase chromatography and tandem mass spectrometry led to the identification of 14 peptides and their parent proteins. Nine of the 14 identified peptides were found to carry 1 or 2 oxidation sites and 5 of the 9 peptides were biotinylated. The significance of this affinity method is that it allows the isolation of oxidized proteins from the rest of the proteome and facilitates their identification. In some cases, it is even possible to identify the site of oxidation. PMID- 15828772 TI - Double-wavelength technique for surface plasmon resonance measurements: basic concept and applications for single sensors and two-dimensional sensor arrays. AB - A new technique for on-line monitoring of analyte binding to sensor surfaces by surface plasmon resonance (SPR) detection is described. It is based on differential measurements using two wavelengths provided by two diode lasers. The technique is as simple and robust as the conventional SPR detection measuring the reflected radiation at fixed incidence angle, but it has the advantage of being nonsensitive to variations of the resonance width and providing essentially higher signal/noise ratios. The paper presents the first four channel prototype system for parallel 2D-monitoring at four different spots. One channel is always used as a reference to compensate temperature fluctuations and nonspecific adsorptions. Calibration with sucrose solutions revealed an absolute sensitivity of Deltan approximately 5 x 10(-6). The new technique is tested with a biotin streptavidin binding and with hybridization/denaturation of DNA. Biotin binding to a streptavidin monolayer is detected with a signal/noise ratio of about 5, which demonstrates the high potential of the new technique for applications in drug discovery. Applications to gene analysis are tested with short oligonucleotides of the sequences used for genotyping human hepatitis C viruses. A selective response to complementary oligonucleotides is observed. The high reproducibility in subsequent cycles of hybridization/denaturation (by formamide or by heating) points out potential applications of the technique in medical diagnostics, food industry, genomics, and proteomics too. PMID- 15828773 TI - Parallel single nucleotide polymorphism genotyping by surface invasive cleavage with universal detection. AB - Large-scale investigations of sequence variation within the human species will provide information about the basis of heritable variation in disease susceptibility and human migration. The surface invader assay (an adaptation of the invasive cleavage reaction to an array format) is capable of exquisitely sensitive and specific detection of genetic variation. It is shown here that this genotyping technology can be multiplexed in a DNA array format, permitting the parallel analysis of a panel of single nucleotide polymorphisms (SNPs) directly from an unamplified genomic DNA target. In addition, a "universal" mode of detection was developed that makes use of a mixture of degenerate templates for DNA ligation to the surface-bound cleaved oligonucleotides and thereby makes this strategy amenable to any desired SNP site or combination of SNP sites, without regard to their particular DNA sequences. This approach was demonstrated on a proof-of-principle scale using small DNA arrays to genotype 6 SNP markers in the PTPN1 gene and 10 mutations in the cystic fibrosis transmembrane conductance regulator gene. This ability to analyze many different genetic variations in parallel, directly from unamplified human genomic DNA samples, lays the groundwork for the development of high-density arrays able to analyze hundreds of thousands or even millions of SNPs. PMID- 15828774 TI - Statistical evaluation of internal and external mass calibration laws utilized in fourier transform ion cyclotron resonance mass spectrometry. AB - The statistical evaluation of two common and three new calibration laws utilized in Fourier transform ion cyclotron resonance mass spectrometry are presented. Electrospray ionization was used to prepare a series of mass spectra of ammonium adducted polypropylene glycol (PPG) with an average molecular weight of 1000 Da. The singly charged PPG-1000 oligomers allowed for the description of a broad range of m/z and abundance values within each mass spectrum. The hexapole accumulation time was varied to afford a range of total ion abundance values of about an order of magnitude. To examine each of the calibration laws, we utilized cross-validation both "within-spectrum" and "between-spectra" for internally and externally calibrated data, respectively. In addition, we used t-statistics to ensure that each calibration coefficient was statistically significant and necessary to accurately describe the variation in the data. In comparison to commonly used calibration laws for internal calibration, our new calibration law based on multiple linear regression offered a 2-fold improvement in mass measurement accuracy (MMA). In comparison to external calibration laws without automatic gain control, our new calibration law using multiple regression improved the MMA by >10-fold; this improvement would increase further as the dynamic range of the measurement increases (e.g., a biological system). For both our internal and external calibration laws, the median MMA was less than 1 part per-million. Furthermore, we investigate the number of calibrant ions as well as their required m/z range in order to successfully achieve high MMA. PMID- 15828775 TI - Enumeration algorithm for determination of binding constants in capillary electrophoresis. AB - With more accurate simulation models and more efficient algorithms becoming available, the binding constants of an affinity interaction can be obtained from much simpler experiments using capillary electrophoresis. With the enumeration algorithm, all possible combinations of the binding constant and the complex mobility in certain ranges that could result in the experimental migration time of an injected analyte are extracted from a 3-D surface, which depicts the migration times resulting from different values of the binding constant and the mobility of the complex formed between the interacting pair, to form a 2-D curve. When the experimental conditions are changed, the analyte migration time will also change. A new 2-D curve can be constructed from another 3-D surface on the basis of the pairs of binding constants and complex mobility values that could result in the new migration time. Because the true binding constant and complex mobility values have to be the same for both experimental conditions under the same temperature, there has to be a point where both 2-D curves will converge. The coordinates of the converging point give the values for a binding constant and a complex mobility that will fit all 2-D curves generated under certain experimental conditions. p-Nitrophenol is used as the analyte, beta-cyclodextrin is used as the additive, and a one-cell model is used to simulate affinity CE. The experimental conditions that can improve the accuracy of the binding constants are discussed. PMID- 15828776 TI - Determination of colchicine residues in sheep serum and milk using high performance liquid chromatography combined with electrospray ionization ion trap tandem mass spectrometry. AB - Colchicine is a naturally occurring alkaloid used in human and veterinary medicine. It shows genotoxicity in in vitro and in vivo systems even at low concentrations. Therefore, no ADI has been established, and colchicine has been included in Annex IV of Council Regulation (EEC) No. 2377/90. No abuse of this drug in intensive livestock farming has yet been reported. However, there may be a natural route of entry for this compound into the food chain when Colchicum autumnale is consumed by animals kept outdoors. To address this concern, we developed and validated a highly sensitive and selective quantitative LC-ESI-MS MS method for the detection of colchicine in sheep serum and milk. For sample pretreatment, all samples were liquid-liquid extracted with phosphate buffer (pH 8.0) and dichloromethane. LC separation was carried out on an RP C18 column employing a 0.5% formic acid/acetonitrile gradient system. The recoveries in both matrixes at a concentration range from 0.0005 to 1 mg/L were >80% with RSDs of <10%. The applicability of the method was demonstrated in serum and the corresponding milk samples from Albanian sheep. After exposure to C. autumnale, the animals showed clinical signs of intoxication. Significant concentrations (up to 1 mg/L) of the drug were found in serum as well as in milk samples. PMID- 15828777 TI - Surface plasmon fluorescence measurements of human chorionic gonadotrophin: role of antibody orientation in obtaining enhanced sensitivity and limit of detection. AB - This paper describes the determination of limits of detection (LODs) of interactions between an antigen, human chorionic gonadotrophin (hCG), and antibodies, anti-alpha-hCG and anti-beta-hCG, using a sandwich assay by surface plasmon field-enhanced fluorescence spectroscopy (SPFS). Randomly biotinylated antibodies were adsorbed onto a structured self-assembled monolayer (SAM) streptavidin matrix, tethered to gold via a SAM consisting of biotinylated thiol molecules interspersed with hydroxyalkanethiol molecules. The influence of the concentration of biotinylated thiol on the binding of biotinylated antibody and its functionality, in terms of its ability to bind to the hCG antigen, was studied. This allowed determination of the optimum biotin-thiol mole fraction in the mixed thiol solution and consequently in the SAM, to maximize binding of hCG of the streptavidin-bound antibody. SPFS studies of the binding of a secondary fluorescently labeled antibody to hCG immobilized on the optimized SAM streptavidin-antibody layer showed that a LOD of hCG of 2 mIU mL(-1) (4 x 10(-12) mol L(-1)) could be realized. The system was further optimized by using a more oriented and organized surface by adsorbing monobiotinylated Fab-hCG in place of the whole antibody. A LOD of 0.3 mIU mL(-1) (6 x 10(-13) mol L(-1)) was achieved for this system. This work illustrates the importance of antibody orientation, both on the planar surface and in terms of position of binding site, in maximizing sensor sensitivity. PMID- 15828778 TI - Determination of thorium and uranium in ultrapure lead by inductively coupled plasma mass spectrometry. AB - A method for the determination of U and Th at sub-ppt levels in high-purity Pb samples using extraction chromatography with ICPMS detection is described. Following acid digestion, uranium and thorium are separated from the lead matrix using UTEVA resin. Sorption and elution procedures were optimized, the potential reusability of the chromatographic resin was evaluated, and a performance comparison between prepacked and freshly prepared UTEVA column was made. Uranium could be eluted with 0.025 M HCl and Th then recovered using 0.5% oxalic acid. Recovery yields for U exceed 80% whereas those for Th were typically 60%. Procedural detection limits of 0.5 and 1.5 pg g(-)(1) were obtained for U and Th, respectively. For purposes of comparison, GD-MS analysis of samples was also performed, yielding results consistent with those generated by ICPMS but with inferior detection power. PMID- 15828779 TI - Radiolytic modification of sulfur-containing amino acid residues in model peptides: fundamental studies for protein footprinting. AB - Protein footprinting based on hydroxyl radical-mediated modification and quantitative mass spectroscopic analysis is a proven technique for examining protein structure, protein-ligand interactions, and structural allostery upon protein complex formation. The reactive and solvent-accessible amino acid side chains function as structural probes; however, correct structural analysis depends on the identification and quantification of all the relevant oxidative modifications within the protein sequence. Sulfur-containing amino acids are oxidized readily and the mechanisms of oxidation are particularly complex, although they have been extensively investigated by EPR and other spectroscopic methods. Here we have undertaken a detailed mass spectrometry study (using electrospray ionization mass spectrometry and tandem mass spectrometry) of model peptides containing cysteine (Cys-SH), cystine (disulfide bonded Cys), and methionine after oxidation using gamma-rays or synchrotron X-rays and have compared these results to those expected from oxidation mechanisms proposed in the literature. Radiolysis of cysteine leads to cysteine sulfonic acid (+48 Da mass shift) and cystine as the major products; other minor products including cysteine sulfinic acid (+32 Da mass shift) and serine (-16 Da mass shift) are observed. Radiolysis of cystine results in the oxidative opening of the disulfide bond and generation of cysteine sulfonic acid and sulfinic acid; however, the rate of oxidation is significantly less than that for cysteine. Radiolysis of methionine gives rise primarily to methionine sulfoxide (+16 Da mass shift); this can be further oxidized to methionine sulfone (+32 Da mass shift) or another product with a -32 Da mass shift likely due to aldehyde formation at the gamma carbon. Due to the high reactivity of sulfur-containing amino acids, the extent of oxidation is easily influenced by secondary oxidation events or the presence of redox reagents used in standard proteolytic digestions; when these are accounted for, a reactivity order of cysteine > methionine approximately tryptophan > cystine is observed. PMID- 15828780 TI - Fluorometric broad-range screening of compounds with affinity for nucleic acids. AB - The potential of a nucleic acid-based optical bioprobe for environmental measurements and drug monitoring is described. The sensor employs the long wavelength intercalating fluorophore TO-PRO-3 (TP3). Compounds that interact with the TP3-DNA complex are indirectly detected by a decrease in the fluorescence intensity. We found that the configuration and length of the DNA dramatically affected the intensity of the fluorescence emitted from the TP3-DNA complex. We compared nucleic acids from different sources and optimized the system for pBR322 plasmid DNA (4363 bp) digested by HindIII restriction endonuclease. This endonuclease has a single recognition site in plasmid pBR322. In the proposed method, we attempt to combine broad-range detection with rapid and simple operation. A fiber-optic capillary fluorescence system was used to analyze toxic aromatic amines, antibiotics, and several kinds of antitumor drugs, using small amounts of sample, down to 10 muL, with a sensitivity comparable to that of current electrochemical methods. The detection limit can be as low as a few ppb or submicromolar. This approach is useful for routine screening in environmental monitoring or for controlling cytotoxic drug administration. The ease of operation and the rapid response allow high-throughput screening. PMID- 15828781 TI - Use of selective TOCSY NMR experiments for quantifying minor components in complex mixtures: application to the metabonomics of amino acids in honey. AB - The application of the traditional methods of multivariate statistics, such as the calculation of principle components, to the analysis of NMR spectra taken on sets of biofluid samples is one of the central approaches in the field of metabonomics. While this approach has proven to be a powerful and widely applicable technique, it has an inherent weakness, in that it tends to be dominated by those chemical species present at relatively higher concentrations. Using a set of commercial honey samples, a comparison of this classical metabonomics approach to one based on the use of the selective TOCSY experiment is presented. While the NMR spectrum of honey and its classical metabonomic analysis is completely dominated by a very few chemical species, specifically alpha-glucose and fructose, the statistical signal carried by minor honey components, such as amino acids, may be accessed using a selective TOCSY-based approach. This approach has the intrinsic virtue that it focuses the statistical analysis on a set of predefined chemical species, which might be chosen for their metabolic significance, and could be composed of either major or minor mixture constituents. Furthermore, the selective TOCSY method allows for more certain chemical identification, acquisition times of approximately 1 min, and accurate quantification of the species contributing to the statistical discriminatory signal. PMID- 15828782 TI - DBDigger: reorganized proteomic database identification that improves flexibility and speed. AB - Database search identification algorithms, such as Sequest and Mascot, constitute powerful enablers for proteomic tandem mass spectrometry. We introduce DBDigger, an algorithm that reorganizes the database identification process to remove a problematic bottleneck. Typically such algorithms determine which candidate sequences can be compared to each spectrum. Instead, DBDigger determines which spectra can be compared to each candidate sequence, enabling the software to generate candidate sequences only once for each HPLC separation rather than for each spectrum. This reorganization also reduces the number of times a spectrum must be predicted for a particular candidate sequence and charge state. As a result, DBDigger can accelerate some database searches by more than an order of magnitude. In addition, the software offers features to reduce the performance degradation introduced by posttranslational modification (PTM) searching. DBDigger allows researchers to specify the sequence context in which each PTM is possible. In the case of CNBr digests, for example, modified methionine residues can be limited to occur only at the C-termini of peptides. Use of "context dependent" PTM searching reduces the performance penalty relative to traditional PTM searching. We characterize the performance possible with DBDigger, showcasing MASPIC, a new statistical scorer. We describe the implementation of these innovations in the hope that other researchers will employ them for rapid and highly flexible proteomic database search. PMID- 15828783 TI - Analysis of perchlorate in human urine using ion chromatography and electrospray tandem mass spectrometry. AB - Because of health concerns surrounding widespread exposure to perchlorate, we developed a sensitive and selective method for quantifying perchlorate in human urine using ion chromatography coupled with electrospray ionization tandem mass spectrometry. Perchlorate was quantified using a stable isotope-labeled internal standard ((18)O(4)-perchlorate) with excellent assay precision (coefficient of variation <5% for repetitively analyzed quality control material). Analytical accuracy was established by blind analysis of certified proficiency testing materials prepared in synthetic urine matrix; calculated amounts deviated minimally from true amounts, with percent differences ranging from 2% to 5%. Selective chromatography and tandem mass spectrometry reduced the need for sample cleanup, resulting in a rugged and rapid method capable of routinely analyzing 75 samples/day. The lowest reportable level (0.025 ng/mL) was sufficiently sensitive to detect perchlorate in all human urine samples evaluated to date, with a linear response range from 0.025 to 100 ng/mL. This selective, sensitive, and rapid method will help elucidate any potential associations between human exposure to low levels of perchlorate and adverse health effects. PMID- 15828784 TI - Capillary electrophoresis and fluorescence anisotropy for quantitative analysis of peptide-protein interactions using JAK2 and SH2-Bbeta as a model system. AB - Fluorescence anisotropy capillary electrophoresis (FACE) and affinity probe capillary electrophoresis (APCE) with laser-induced fluorescence detection were evaluated for analysis of peptide-protein interactions with rapid binding kinetics. The Src homology 2 domain of protein SH2-Bbeta (SH2-Bbeta (525-670)) and a tyrosine-phosphorylated peptide corresponding to the binding sequence of JAK2 were used as a model system. For peptide labeled with fluorescein, the K(d) = 82 +/- 7 nM as measured by fluorescence anisotropy (FA). APCE assays had a limit of detection (LOD) of 100 nM or 12 amol injected for SH2-Bbeta (525-670). The separation time of 4 s, achieved using an electric field of 2860 V/cm on 7-cm long capillaries, was on the same time scale as complex dissociation allowing K(d) (101 +/- 12 nM in good agreement with FA measurements) and dissociation rate (k(off) = 0.95 +/- 0.02 s(-)(1) corresponding to a half-life of 0.73 s) to be determined. This measurement represents a 30-fold higher rate of complex dissociation than what had previously been measurable by nonequilibrium CE analysis of equilibrium mixtures. Using FACE, the protein was detected with an LOD of 300 nM or 7.5 fmol injected. FACE was not used for determining K(d) or k(off); however, this method provided better separation resolution for multiple forms of the protein than APCE. Both methods were found suitable for analysis of cell lysate. These results demonstrate that FACE and APCE may be useful complements to existing techniques for exploring binding interactions with rapid kinetics. PMID- 15828786 TI - Thermodynamic studies of the solvent effects in chromatography on molecularly imprinted polymers. 3. Nature of the organic mobile phase. AB - Experimental isotherm data of the Fmoc-tryptophan (Fmoc-Trp) enantiomers were measured by frontal analysis on a Fmoc-L-Trp imprinted polymer, using different organic mobile phases, in a wide concentration range. The nonlinear regression of the data and the independent calculation of the affinity energy distributions of the two enantiomers allowed the selection of the isotherm model and the determination of the isotherm parameters. The organic solvents studied were acetonitrile (MeCN), methylene chloride, chloroform, and tetrahydrofuran (THF), all in the presence of the same concentration of acetic acid, used as an organic modifier. It was found that the highest overall affinity and enantiomeric selectivity were obtained in MeCN, which is also the solvent used in the polymerization. In the other solvents, the overall affinity decreases with increasing hydrogen-bonding ability of the solvents but not the enantiomer selectivity. In MeCN, three types of adsorption sites coexist for the two enantiomers on the MIP. The highest energy sites for Fmoc-L-Trp in MeCN are inactive in CH(2)Cl(2), CHCl(3), and THF, and only two types of sites were identified in these solvents. Increasing the acetic acid concentration from 0.2 to 0.9 M causes a large decrease in the association constant of the highest energy sites in CH(2)Cl(2), CHCl(3), and THF but not in MeCN. The overall affinity of Fmoc-L-trp in CH(2)Cl(2), CHCl(3), and THF is dominated by adsorption on the lowest energy sites, the most abundant ones. In contrast, in MeCN, the overall affinity of Fmoc-L-Trp is dominated by adsorption on the highest energy sites, the least abundant sites. In CH(2)Cl(2), CHCl(3), and THF, the number of each type of sites increases with decreasing hydrogen-bonding ability of the solvents while the association constant of the corresponding sites does not change significantly. PMID- 15828785 TI - Cytometry and velocimetry on a microfluidic chip using polyelectrolytic salt bridges. AB - This paper reports a polyelectrolytic salt bridge-based electrode (PSBE), which is a key embedded unit in a microchip device that can size-selectively count microparticles and measure their velocities. The construction of salt bridges at specific locations within a microfluidic chip enables dc-driven electrical detection to be performed successfully. This is expected to be a competitive alternative to the optical methods currently used in conventional cell sorters. The PSBEs were fabricated by irradiating ultraviolet light over a patterned mask on the parts of interest, which were filled with an aqueous monomer solution containing diallyldimethylammonium chloride. A pair of such PSBEs was easily formed at the two lateral branches perpendicular to the main microchannel and was found to be very useful for dc impedometry. The human blood cells as well as the fluorescent microbeads passing between the two PSBEs produced impedance signals in proportional to their size. The information about the velocity of a microparticle was extracted from a doublet of the dc impedance signals, which were generated when cells or microbeads sequentially passed through two PSBE pairs separated from each other by a fixed distance. The plot of peak amplitude versus velocity of the moving microbeads and cells indicated only a slight correlation between the size and the velocity, which means that the peak amplitude of the dc impedance signals alone can provide information about the size of the cells in a mixture. The experimental results showed a screening rate of over 1000 cells s(-1) and a velocity of the cells of over 100 mm s(-1). Compared with the previously suggested electrical detection system based on metal electrodes, the sensitivity and selectivity in cell detection were remarkably improved. In addition, the detection unit including the operating circuit became innovatively simple and the whole device could be miniaturized. PMID- 15828787 TI - Nondestructive determination of solids and carotenoids in tomato products by near infrared spectroscopy and multivariate calibration. AB - Tomato is an important player in the agricultural market. It is the second most consumed vegetable in the world and is a source of important micronutrients such as lycopene and beta-carotene. Recent research has demonstrated that these carotenoids can act as free-radical quenchers in the body and prevent aging, tissue damage, heart disease, and certain cancers. Besides these microcomponents, tomato is composed of soluble and insoluble solids. In industry, these solids govern factory yield and play a major role in the tomato trade. Nowadays, standard methods for determining tomato solids and carotenoids are time and labor consuming. In this work, we present the development of a simultaneous and nondestructive method for determining total and soluble solids, as well as lycopene and beta-carotene, in tomato products by near-infrared spectroscopy. PLS 1 was the calibration technique chosen. For spectra preprocessing, MSC and second derivative were applied. As variable selection techniques, the correlogram cutoff, the successive projections algorithm, the dimension wise selection, and spectra splitting approach were applied. Best models presented satisfactory prediction abilities evaluated through its RMSEP and r values: total solids 0.4157, 0.9998; soluble solids 0.6333, 0.9996; lycopene 21.5779, 0.9996; beta carotene 0.7296, 0.9981. PMID- 15828788 TI - Partitioning model for competitive host-guest complexation in ESI-MS. AB - Mathematical models based on equilibrium partitioning theory were developed to relate ion abundances produced by electrospray ionization mass spectrometry with solution concentrations of complexes resulting from competitive host-guest binding. Through modeling the possible equilibria in the electrospray droplets, including the partitioning between the droplet surface and interior that dictate what ions are generated by an electrospray, the factors responsible for distorting the distribution of ions from their solution concentrations were evaluated. Experiments with crown ether-alkali metal complexes confirm the validity of the models and yield a greater understanding of the behavior of host guest complexes in ESI-MS, allowing for more accurate and less trouble-prone measurements of solution binding interactions. PMID- 15828789 TI - Single-molecule observation and chromatography unified by Levy process representation. AB - A renewed stochastic model of chromatography is proposed able to establish a conceptual link between the single-molecule dynamics observations in a given chromatographic system and chromatographic experiment results in the same media. The chromatographic peak is expressed through its Fourier transform as a function of the experimental sorption time distribution. The pertinent numerical procedure necessary for obtaining the chromatographic peak is described, and the numerical programming code is given. Two types of cases were considered, the first one where the sole discrete sorption time distribution is available, and the second one where the adsorption mechanism is made of a mixture of continuous and discrete sorption time distributions. The method is applied to experimental data found in the literature by determining typical chromatographic peak shapes on the basis of the experimental interface adsorption data. This renewed stochastic approach is based on the so-called Levy canonical description of stochastic processes and appears to be the most general basis for handling separation processes from a stochastic point of view. PMID- 15828790 TI - Characterization of high-molecular-weight sulfur-containing aromatics in vacuum residues using Fourier transform ion cyclotron resonance mass spectrometry. AB - Millions of tons of vacuum residues are produced in refineries every year and could be a potentially valuable resource for generating electricity and has possible application as heating and marine fuel. In this work, the polycyclic aromatic sulfur compounds (PASHs) from the aromatic fraction of vacuum residue before and after partial hydrodesulfurization (HDS) were derivatized by methylation to the methylsulfonium salts. Fourier transform ion cyclotron resonance mass spectrometry provided high-resolution data on these high-molecular weight sulfur compounds. Compounds containing one and two S atoms were found to dominate, with masses up to ca. 900 Da. Classification according to hydrogen deficiency and the number of heteroatoms showed extensive series of homologues for double bond equivalents from 5 to 20. The sulfur-containing aromatics were separated using a palladium(II) complex as a liquid chromatographic phase into two compound groups: one containing compounds with an unconjugated thiophene ring and another with a condensed thiophene ring. This, combined with the mass spectrometry (MS) data, allows for the identification of several parent structures. Partial HDS removed primarily compounds with one S atom, whereas those with two S atoms were largely unaffected. PMID- 15828791 TI - Cloud point extraction coupled with microwave or ultrasonic assisted back extraction as a preconcentration step prior to gas chromatography. AB - Cloud point extraction of nonionic and anionic surfactants was applied as a preconcentration step prior to gas chromatography. No cleanup step preceded chromatographic analysis. The obtained surfactant-rich phase was treated with water-immiscible solvents, and the target analytes were back extracted by short term microwave application or ultrasonication. A mixture of six PAHs (naphthalene, acenaphthene, fluorene, anthracene, fluoranthene, pyrene) was used as test compounds. The obtained detection limits were in the microgram per liter area. Recoveries of spiked water and soil samples ranged between 92 and 105% while analysis of certified reference materials gave results in good agreement with the certified values. Under the optimum experimental conditions, there was no interference or blocking of the column. According to our results, this approach presents a convenient solution to the up-to-date problem of combining gas chromatography with micellar cloud point extraction. PMID- 15828792 TI - DNA biosensor for detection of Helicobacter pylori using phen-dione as the electrochemically active ligand in osmium complexes. AB - A surface-based method for the study of the interactions of DNA with redox-active 1,10-phenantroline-5,6-dione (phen-dione) osmium complexes is described. The study was carried out using gold electrodes modified with DNA via adsorption and [Os(bpy)(2)(phe-dione)](3+/2+) (bpy = 2,2'-bipyridyl) or [Os(phen)(2)(phen dione)](3+/2+) (phen = 1,10-phenantroline) as electrochemical reported molecules. The method, which is simple and reagent-saving, allows the accumulation of osmium complexes within the DNA layer. The amount of osmium complex bound by the adsorbed layer of DNA was determined from the voltammetric charge associated with the osmium redox process of the immobilized metal complex. The quinone moiety of the phen-dione ligand was useful as an indicator for electrochemical DNA sensing because of its redox response at low potentials. A thiol-linked single-stranded Helicobacter pylori DNA probe was immobilized, through S-Au bonds on to a gold electrode (density of modification 86 pmol/cm(2)). Following hybridization with the complementary DNA sequence, the osmium complex was electrochemically accumulated within the double-stranded DNA layer. Electrochemical detection was performed by differential pulse voltammetry over the potential range where the quinone moiety was redox active (i.e., at very low potentials, -0.020 V vs SSCE); with this approach, a sequence of the H. pylori could be quantified over the range from 5 to 20 pmol with a linear correlation of r = 0.9888 and a detection limit of approximately 6 pmol. PMID- 15828793 TI - Enzyme kinetics in acoustically levitated droplets of supercooled water: a novel approach to cryoenzymology. AB - The rate of the alkaline phosphatase-catalyzed hydrolysis of 4 methylumbelliferone phosphate was measured in acoustically levitated droplets of aqueous tris (50 mM) at pH 8.5 at 22 +/- 2 degrees C and in supercooled solution at -6 +/- 2 degrees C. At 22 degrees C, the rate of product formation was in excellent agreement with the rate observed in bulk solution in a cuvette, indicating that the acoustic levitation process does not alter the enzyme activity. The rate of the reaction decreased 6-fold in supercooled solution at -6 +/- 2 degrees C. The acoustic levitator apparatus is described in detail. PMID- 15828794 TI - Real-time detection of hyperosmotic stress response in optically trapped single yeast cells using Raman microspectroscopy. AB - Living cells survive environmentally stressful conditions by initiating a stress response. We monitored changes in the Raman spectra of optically trapped Saccharomyces cerevisiae yeast cell under normal, heat-treated, and hyperosmotic stress conditions. It is shown that when glucose was used to exert hyperosmotic stress, two chemical substances-glycerol and ethanol-can be monitored in real time in a single cell. PMID- 15828795 TI - Subpicomolar sensing of delta-opioid receptor ligands by molecular-imprinted polymers using plasmon-waveguide resonance spectroscopy. AB - Here we report, for the first time, the formation of a biomimetic covalently imprinted polymeric sensor for a target ligand, the delta-opioid G-protein coupled receptor agonist DPDPE, which reproducibly exhibits subpicomolar binding affinity in an aqueous environment. In addition to having a well-defined and homogeneous binding site, the imprinted polymer template is quite stable to storage in both the dry and wet states and has at least 6 orders of magnitude higher affinities than exhibited by similar peptide-based molecular-imprinted polymers (MIPs) thus far. A highly sensitive optical detection methodology, plasmon-waveguide resonance spectroscopy, was employed, capable of measuring binding in real time and discriminating between ligand molecules, without requiring labeling protocols (fluorophores or radioisotopes). The DPDPE-imprinted polymer showed a broad structure-activity relationship profile, not unlike that found for protein receptors. Such sensitivity and robustness of MIPs suggests potential applications ranging from biowarfare agent detection to pharmaceutical screening. PMID- 15828796 TI - SIMPLISMA and ALS applied to two-way nonlinear wavelet compressed ion mobility spectra of chemical warfare agent simulants. AB - Ion mobility spectrometry is a rapid scanning measurement method for which compression methods that facilitate the handling of large collections of data are beneficial. Peak distortion in reconstructed ion mobility spectra from linear wavelet compression is problematic in that artifact peaks may cause false positive alarms. Peak shifting also may cause false alarms if target peaks shift out of or interfering peaks shift into detection windows. Nonlinear wavelet compression (NLWC) preserves peak shape and can lessen the degree of distortion, shifting, and artifact peaks in the reconstructed spectra. NLWC was applied to achieve high compression and fidelity in the reconstructed spectra. Another benefit is that NLWC improves signal-to-noise ratios and thus the models built from compressed data are improved. By compressing both the drift time order and the spectrum acquisition order, greater compressions maybe achieved. A two-way nonlinear wavelet compression method that incorporates alternating least squares (2W-NLWC-ALS) algorithm was devised by applying ALS to partially reconstructed wavelet coefficients generated from two-way NLWC. The number of components in a data set can be determined automatically using ASIMPLISMA. The smaller ALS models are saved as the final compressed data and can be used to reconstruct the entire data set efficiently without maintaining the compressed wavelet coefficient matrix of the original data set. The 2W-NLWC-ALS algorithm provides greater compression ratios compared to regular wavelet compression and interpretable models. Using this method, large volumes of data can be acquired and easily evaluated through a simple compressed model. A compression ratio of 510 ppm, root mean-square error (E(RMS)) of 6.3 mV (full-scale signal is usually 1 V or larger), and relative root-mean-square error (RE(RMS)) of 1.62% were achieved for data sets collected by CAM. A compression ratio of 46 ppm, E(RMS) of 9.2 mV, and RE(RMS) of 0.42% were achieved for data sets collected with an ITEMISER instrument. The 2W-NLWC-ALS algorithm is an efficient compression method that provides the benefits of a simple model. PMID- 15828797 TI - Monitoring food quality by microfluidic electrophoresis, gas chromatography, and mass spectrometry techniques: effects of aquaculture on the sea bass (Dicentrarchus labrax). AB - Monitoring food quality is a critical task for analytical chemistry and an important way to preserve human health. Fish is a valuable source of highly digestible proteins and contains large amounts of polyunsaturated fatty acids and fat-soluble vitamins. Since the world's wild fish stocks are limited, farmed fish is nowadays proposed as an alternative to consumers. It is now emerging that the fish muscle protein content is assuming great importance from an aquaculture perspective. Many data have been collected on the physiology and biochemistry of fish muscle, but few proteomic studies are available on farmed fish. Application of proteomics to aquaculture may play a key role in the development of new farming strategies. In this paper, a proteomic approach based on SDS-PAGE separation of proteins, in situ protein hydrolysis, de novo sequencing of peptides by MALDI and ESI MS(2), protein identification, and relative quantitation of protein by denaturing capillary electrophoresis was coupled with the determination of fatty acids and metal ions content by GM-MS and ICPMS in farmed and wild sea bass filet. Our results show that aquaculture could induce significant chemical and biochemical differences in fish muscle that may have an impact on food quality. PMID- 15828798 TI - Gram-scale synthesis and biofunctionalization of silica-coated silver nanoparticles for fast colorimetric DNA detection. AB - A direct silica-coating method has been developed for the gram-scale synthesis of well-dispersed Ag@SiO(2) nanoparticles. Subsequent surface functionalization via the well-established silica surface chemistry provided arching points for straightforward bioconjugation with amino-terminated oligonucleotides. Fast hybridization kinetics of the resulting robust oligo-modified Ag@SiO(2) nanoprobes with complementary target oligonucleotides render themselves very useful for the fast colorimetric DNA detection based on the sequence-specific hybridization properties of DNA. Additionally, the reliable protocols developed in this study for preparing and functionalizing Ag@SiO(2) nanoparticles can be readily extended to other silica-coated nanoparticles, which can also provide a specific platform for the covalent attachment of biomolecules such as amino-rich proteins, enzymes, or amino-terminated oligonucleotides for diverse bioapplications. PMID- 15828799 TI - Microelectrode array microscopy: investigation of dynamic behavior of localized corrosion at type 304 stainless steel surfaces. AB - Scanning electrochemical microscopy (SECM) and a recently developed microelectrode array microscope have been used to study localized corrosion and electron-transfer characteristics of native oxide layers of type 304 stainless steels. The I-/I3- redox couple was employed as a mediator and allowed sensitive detection of oxide breakdown events. In solutions containing I-, a signal at the microelectrode was observed on type 304 stainless steel surfaces at active pitting corrosion sites. Under conditions where pitting corrosion occurs, SECM was used to track the temporal characteristics of the reaction in a spatial manner. However, because of the time required to create an image, much of the temporal information was not obtained. To improve the temporal resolution of the measurement, microelectrode array microscopy (MEAM) was developed as a parallel method of performing SECM. The demonstration shown reveals the potential of MEAM for analysis of surface chemistry on temporal and spatial domains. PMID- 15828800 TI - Four-way data coupled to parallel factor model applied to environmental analysis: determination of 2,3,7,8-tetrachloro-dibenzo-para-dioxin in highly contaminated waters by solid-liquid extraction laser-excited time-resolved Shpol'skii spectroscopy. AB - This article reports the first application of parallel factor analysis to high order instrumental data generated from Shpol'skii matrixes at liquid helium temperature. Third-order data arrays-consisting of excitation modulated wavelength time matrixes-are collected with the aid of a cryogenic fiber-optic probe, a tunable dye laser, and a multichannel system for phosphorescence detection. The multidimensional data formats are applied to the analysis of 2,3,7,8-tetrachloro-dibenzo-para-dioxin in water samples. The experimental procedure is rapid and environmentally friendly. Complete sample analysis is accomplished in less than 15 min with only 100 muL of organic solvent (n heptane). The feasibility to directly determine parts-per-trillion concentration levels of the target compound is demonstrated with heavily contaminated samples of unknown composition. The limits of detection, estimated from calibrations based on the univariate method and parallel factor analysis, are 0.060 and 0.092 ng.mL(-1), respectively; both referred to a 100 mL water sample. PMID- 15828801 TI - Minimally destructive analysis of aluminum alloys by resonance-enhanced laser induced plasma spectroscopy. AB - Aluminum alloys were analyzed for minor components and trace impurities using double pulse resonance-enhanced laser-induced plasma spectroscopy. The first laser pulse at 532 nm ablated the sample. The second laser pulse at 396.15 nm resonantly excited the Al atoms to rekindle the plasma plume. Emissions from Mg, Cu, Si, and Na were observed. At laser energies below the damage threshold, the analyte emissions were already orders of magnitude above the background noise. Nonresonant probes of comparable sensitivity would melt and deform the sample surface. Because of the lower etch rate of resonant probes, depth profiling at nanometer resolution was possible. Using this method, the variation of [Na] with depth was measured for high-purity samples. In contrast, nonresonant probes required 5 times the fluence and proportionally poorer resolution. Worse yet, the associated heating and laser remelting modified the [Na] profile. PMID- 15828802 TI - 5-Hydroxytryptophan as a precursor of a catalyst for the oxidation of NADH. AB - Following oxidation of 5-hydroxytryptophan (5-HTPP) at a pyrolytic graphite electrode at pH 7.5, two quasi-reversible redox couples emerge at -0.170 and +0.032 V, respectively, due to oxidation products strongly adsorbed to the electrode surface. These redox processes have been electrochemically and kinetically characterized in terms of the dependence of the formal potential (E degrees ') with pH, variation of the current density with scan rate, operational stability, and electron-transfer rate constant (k(s)). The wave centered at +0.032 V could mediate the oxidation of NADH, exhibiting a strong and persistent electrocatalytic response. A quinone-imine structure has been proposed as the electrocatalytically active species. The kinetics of the reaction between the mediator and NADH has been characterized via rotating disk electrode voltammetry, and it has been found that the rate constant for the reaction is dependent on the solution concentration of NADH. 5-HTPP modified electrodes could be employed in the amperometric detection of NADH with a limit of detection in the nanomolar range. Moreover, 5-HTPP modified electrodes retain their electrocatalytic activity for at least one week. The potential application of these electrodes to amperometric biosensor is demonstrated. PMID- 15828803 TI - Electrochemical method for the detection of lipase activity. AB - A novel electrochemical technique for the general assay of lipase activity is described. The method utilizes a solid-supported lipase substrate, which is formed by dripping and drying a small amount of an ethanol solution of 9-(5' ferrocenylpentanoyloxy)nonyl disulfide (FPONDS) onto gold modified by a hexanethiol self-assembled monolayer. The redox ferrocene group of FPONDS generates the electrochemical signal, the intensity of which is proportional to the number of FPONDS molecules at the interface. Electrochemical and surface enhanced infrared absorption spectroscopic data, as well as control experiments with an engineered, deactivated mutant enzyme, demonstrate that the wild-type lipase from Thermomyces lanuginosus is capable of cleaving the ester bonds of FPONDS molecules via an enzymatic hydrolysis mechanism, which includes the adsorption of the lipase onto the substrate surface. The hydrolysis liberates the ferrocene groups from the interface triggering a decay of the electrochemical redox signal. The rate of the electrochemical signal decrease is proportional to the lipase activity/concentration. These data suggest a general method for the direct measure of enzymatic activity of lipases. PMID- 15828804 TI - One-step homogeneous immunoassay for small analytes. AB - We have developed a one-step, homogeneous noncompetitive immunoassay for small analytes using recombinant antibodies and morphine as the model analyte. A highly specific antibody against the immune complex (IC) formed between an anti-morphine antibody and morphine was selected from a naive scFv phage display library. The in vitro phage library selection procedure avoids the difficulties associated with the production of anti-IC antibodies by animal immunization. The anti morphine and the anti-IC antibodies were labeled with a pair of fluorescence resonance energy transfer (FRET) fluorophores. In the FRET assay the labeled antibodies were incubated with saliva samples spiked with morphine, codeine, or heroin. Within 2 min, 5 ng/mL morphine, which is clearly under the recommended cutoff level, was detected without cross-reactivity to codeine or heroin. This assay principle is also widely applicable to other small analytes. PMID- 15828805 TI - Synthesis, characterization, and application of Eu(III), Tb(III), Sm(III), and Dy(III) lanthanide chelate nanoparticle labels. AB - Preparation and characterization of europium(III), terbium(III), samarium(III), and dysprosium(III) polystyrene nanoparticle labels with lanthanide-specific fluorescence properties has been presented. Emulsion copolymerization of styrene and acrylic acid was used to synthesize uniform-sized nanoparticles approximately 45 nm in diameter. Europium(III) and samarium(III) lanthanides were chelated with 2-naphthoyltrifluoroacetone and trioctylphosphine oxide to dye the spherical particles, whereas terbium(III) and dysprosium(III) chelate complexes contained a newly synthesized ligand, 4-(2,4,6-tridecyloxyphenyl)pyridine-2,6-dicarboxylic acid. The fluorescence properties of the four lanthanides-including a wide Stokes shift, a narrow emission peak, and long fluorescence lifetime-were retained despite the incorporation into the nanoparticles. Furthermore, the nanoparticles, containing more than 1000 lanthanide chelates, were detectable at label concentrations 3 orders of magnitude lower than the corresponding soluble lanthanide chelate labels. The applicability of the labels prepared was demonstrated by a heterogeneous sandwich-type immunoassay for human prostate specific antigen, where the lowest limits of detection of 1.6, 2.4, 10.1, and 114.2 ng/L were achieved using europium(III), terbium(III), samarium(III), and dysprosium(III) nanoparticles, respectively. The spectral and functional properties of the lanthanide-embedded polystyrene nanoparticles developed here suggest that the technology is applicable for high-sensitivity multicolor assays. PMID- 15828806 TI - Integrated sample processing system involving on-column protein adsorption, sample washing, and enzyme digestion for protein identification by LC-ESI MS/MS. AB - An automated system has been developed for protein identification using mass spectrometry that incorporates sample cleanup, preconcentration, and protein digestion in a single stage. The procedure involves the adsorption of a protein or a protein mixture from solution onto a hydrophobic medium that is contained within a microcolumn. The protein is digested while still bound to the hydrophobic support. The peptides are then eluted from surface digestion to an electrospray ionization mass spectrometer for detection and sequencing. The entire system is fully automated wherein the mass spectrometer is collecting data continuously. We demonstrate that this system is capable of identifying standard protein samples at concentrations down to 100 nM. Further development of this technique may offer a potential solution for proteomics applications that require unattended operation, such as on-line monitoring and identification of microorganisms on the basis of the detection of their protein biomarkers. PMID- 15828807 TI - Microscope objective for large-angle fluorescence used for rapid detection of single nucleotide polymorphisms in DNA hybridization. AB - A new type of microscope objective is used for the rapid detection of sequence dependent affinity variations in DNA hybridization. We demonstrate that by performing probe/target hybridization on coverslips at room temperature terminal SNPs (single nucleotide polymorphisms) can be detected within seconds. The study of weak pair interaction, such as the association of very short DNA oligomers, requires the use of high analyte concentrations of both partners to generate a detectable amount of associated pairs. The background of high concentrations of unbound fluorescing analyte can easily hide the low signal of a weakly affine reaction and makes association extremely difficult to detect. Fluorescence detection is a powerful approach to analyze minute amounts of material, even single molecules, but it is usually limited to rather low concentrations. This limitation is now overcome due to the new type of microscope objective, which produces an extremely small detection volume at a water/glass interface. PMID- 15828808 TI - Supporting electrolyte and solvent effects on single-electron double layer capacitance charging of hexanethiolate-coated Au140 nanoparticles. AB - Sequential injections of single electrons (or holes) into the cores of Au(140) hexanethiolate monolayer-protected clusters (MPCs) occur at measurably different electrochemical potentials owing to the extremely small (subattofarad) values of the single MPC capacitance (C(MPC)) of the nanoparticle. The potential increment for each sequential injection is DeltaV = e/C(MPC). The dependence of DeltaV on the concentration of supporting electrolyte (from 1 to 100 mM), measured using square wave voltammetry, is shown to be caused, primarily, by changes in the diffuse double layer component (C(DIFFUSE)) of C(MPC). The dependence of C(DIFFUSE) on r(core), the radius of the nanoparticle, is considered. Additionally, significant changes in the magnitude of the compact double layer component (C(COMPACT), equivalent to the Stern layer) of C(MPC) were induced by adding hydrophobic solvent components such as hexane or dodecane or by introducing hydrophobic electrolyte ions (tetrabutyl-, tetrahexyl-, and tetraoctylammonium, perchlorate, and tetraphenylborate). These changes are interpreted as specific solvation and ion penetration of the hexanethiolate monolayer. For brevity we will refer to these phenomena as solvation/penetration phenomena. PMID- 15828809 TI - High-performance quenchometric oxygen sensors based on fluorinated xerogels doped with [Ru(dpp)3]2+. AB - By using hybrid xerogels that are composed of alkyl and perfluoroalkyl ORMOSILs (organically modified silicates) doped with the luminophore tris(4,7'-diphenyl 1,10'-phenanthroline) ruthenium(II), we have produced highly sensitive O2 sensors (i.e., I(N2)/I(O2) = 35 +/- 4) with linear calibration curves that exhibit less than 2% drift over 6 months. PMID- 15828810 TI - Cytochrome p450 in silico: an integrative modeling approach. PMID- 15828811 TI - Discovery of S-phase arresting agents derived from noscapine. AB - Analogues of the natural product noscapine were synthesized, and their potential as antitumor agents were examined. The discovery of a novel regio- and stereoselective O-demethylation led to the synthesis of several O-alkylated analogues that induced an unexpected S-phase arrest of mammalian cells. Compound 4a was the most potent analogue inhibiting cell proliferation at an EC(50) of 1.9 microM. PMID- 15828812 TI - Novel and potent 17beta-hydroxysteroid dehydrogenase type 1 inhibitors. AB - Structure-based drug design using the crystal structure of human 17beta hydroxysteroid dehydrogenase type 1 (17beta-HSD1) led to the discovery of novel, selective, and the most potent inhibitors of 17beta-HSD1 reported to date. Compounds 1 and 2 contain a side chain with an m-pyridylmethyl-amide functionality extended from the 16beta position of a steroid scaffold. A mode of binding is proposed for these inhibitors, and 2 is a steroid-based 17beta-HSD1 inhibitor with the potential for further development. PMID- 15828813 TI - Effects of 5'-phosphate derivatives of 2-hexynyl adenosine and 2-phenylethynyl adenosine on responses of human platelets mediated by P2Y receptors. AB - Newly synthesized mono-, di-, and triphosphate of 2-alkynyl adenosines showed very different behavior in human platelet P2Y receptor models, according to the different alkynyl chains. In fact, 2-hexynyladenosine di- (5) and triphosphate (7) induced platelet shape change and aggregation and inhibited PGE(1)-induced increase in platelet cyclic AMP. On the contrary, the corresponding 2 phenylethynyladenosine di- (6) and triphosphate (8) did not induce platelet shape change or aggregation, but inhibited platelet aggregation induced by ADP. PMID- 15828815 TI - Synthesis and biological evaluation of 11-substituted 6 aminobenzo[c]phenanthridine derivatives, a new class of antitumor agents. AB - The synthesis of 11-substituted 6-amino-11,12-dihydrobenzo[c]phenanthridines and 11-substituted 6-aminobenzo[c]phenanthridines through an efficient method is described. The antiproliferative activity of selected compounds against a wide panel of tumor cell lines was tested in the in vitro anticancer screening and the in vivo hollow fiber assay of the National Cancer Institute. Several compounds turned out to exhibit considerable cytotoxicity for tumor cells. For the study of structure-activity relationships different substituents were introduced in the 11 position. Compounds with methoxyphenyl substituents tended to show the highest potency. Several compounds exhibited noteworthy antitumor activity with GI(50) values across all cell lines <1 microM. 6-Amino-11-(3,4,5 trimethoxyphenyl)benzo[c]phenanthridine perchlorate was the most potent agent in the NCI's in vivo hollow fiber assay. Most of the tested compounds showed a remarkable selectivity for leukemia, breast cancer, and prostate cancer. PMID- 15828814 TI - Synthesis and monoamine transporter binding properties of 3beta-(3',4' disubstituted phenyl)tropane-2beta-carboxylic acid methyl esters. AB - 3beta-(3'-Methyl-4'-chlorophenyl)tropane-2-carboxylic acid methyl ester (3b, RTI 112) is a 3-phenyltropane analogue that has high affinity for both the dopamine and serotonin transporters (DAT and 5-HTT, respectively). Compound 3b shows significant reduction of cocaine self-administration in rhesus monkeys, yet fails to maintain robust drug self-administration. PET studies revealed that unlike more DAT selective analogues such as GBR 12 909 and 3-(4-chlorophenyl)tropane-2 carboxylic acid phenyl ester (RTI-113), 3b shows no detectible DAT occupancy when dosed at its ED(50) for reduction of cocaine self-administration. In contrast, it highly occupies the 5-HTT at this dose. In this study, we report the synthesis and monoamine transporter binding potency of several new 3-(3',4'-disubstituted phenyl)tropane-2-carboxylic acid methyl esters (3c-k), which have binding properties very similar to 3b. With the exception of the 3',4'-dimethyl analogue 3k, all of the compounds possess subnanomolar IC(50) and K(i) values at the DAT and 5-HTT, respectively. The 3'-chloro-4'-bromo analogue 3e (IC(50) = 0.12 nM) and the 3'-bromo-4'-iodo analogue 3i (K(i) = 0.14 nM) are the most potent analogues at the DAT and 5-HTT, respectively. These compounds will be useful to further characterize the highly interesting behavioral profile of 3b. PMID- 15828816 TI - N-terminal sugar conjugation and C-terminal Thr-for-Thr(ol) exchange in radioiodinated Tyr3-octreotide: effect on cellular ligand trafficking in vitro and tumor accumulation in vivo. AB - For effective targeting of somatostatin receptor (sst) expressing tumors by radiolabeled octreotide analogues, high ligand uptake into sst-positive cells is mandatory. To optimize it, two modifications have been introduced into [(125)I]Tyr(3)-octreotide ([(125)I]TOC): C-terminal Thr-for-Thr(ol) exchange (leading to Tyr(3)-octreotate (TOCA)) and N-terminal derivatization with different carbohydrates. Both have significant impact on radioligand uptake into sst(2)-expressing cells in vitro and in vivo. Glucose conjugation via Amadori reaction by itself led to improved tumor uptake of [(123)I]Gluc-TOC in vivo, which is based on an enhancement of peptide internalization despite a reduction in receptor affinity. In the case of the doubly modified analogues [(123)I]Gluc TOCA, [(123)I]Gluc-S-TOCA, and [(123)I]Gal-S-TOCA, a cumulative effect of both structural modifications was observed, leading up to a 5-fold increased uptake of these compounds in sst-expressing tumors compared to [(125)I]TOC. Thus, glycosylation with small carbohydrates was found to be a suitable tool to enhance receptor-mediated uptake of radiolabeled octreotide analogues into sst-positive malignancies, leading to tracers with excellent characteristics for in vivo sst imaging applications. PMID- 15828817 TI - Effects of flavonoids on cell proliferation and caspase activation in a human colonic cell line HT29: an SAR study. AB - A library of 42 natural and synthetic flavonoids has been screened for their effect on cell proliferation and apoptosis in a human colonic cell line (HT-29). Examples of different classes of flavonoids have been screened, and the effects of hydroxylation, methoxylation and/or C-alkylation at various positions in the A and B-rings have been assessed. Flavones and flavonols possess greater antiproliferative activity than chalcones and flavanones. With respect to structural modification of flavonoids, C-isoprenylation was by far the most effective, with substitution at the 8-position and longer chains, such as geranyl giving the best results. Finally, most compounds that significantly reduced cell survival also increased caspase activity, suggesting that at least part of their antiproliferative activity might be attributable to an apoptotic response. PMID- 15828818 TI - In vitro metabolism of phenoxypropoxybiguanide analogues in human liver microsomes to potent antimalarial dihydrotriazines. AB - Phenoxypropoxybiguanides, such as 1 (PS-15), are prodrugs analogous to the relationship of proguanil and its active metabolite cycloguanil. Unlike cycloguanil, however, 1a (WR99210), the active metabolite of 1, has retained in vitro potency against newly emerging antifolate-resistant malaria parasites. Unfortunately, manufacturing processes and gastrointestinal intolerance have prevented the clinical development of 1. In vitro antimalarial activity and in vitro metabolism studies have been performed on newly synthesized phenoxypropoxybiguanide analogues. All of the active dihydrotriazine metabolites exhibited potent antimalarial activity with in vitro IC(50) values less than 0.04 ng/mL. In vitro metabolism studies in human liver microsomes identified the production of not only the active dihydrotriazine metabolite, but also a desalkylation on the carbonyl chain, and multiple hydroxylated metabolites. The V(max) for production of the active metabolites ranged from 10.8 to 27.7 pmol/min/mg protein with the K(m) ranging from 44.8 to 221 microM. The results of these studies will be used to guide the selection of a lead candidate. PMID- 15828819 TI - Selective estrogen receptor modulators in the ruthenocene series. Synthesis and biological behavior. AB - A series of ruthenocene derivatives, 1-[4-(O(CH(2))(n)()N(CH(3))(2))phenyl]-1-(4 hydroxyphenyl)-2-ruthenocenylbut-1-ene, with n = 2-5, based on the structure of the breast cancer drug tamoxifen has been prepared. These compounds were obtained, via a McMurry cross-coupling reaction, as a mixture of Z and E isomers that could not be separated by HPLC. The relative binding affinity values for estrogen receptor alpha (ERalpha) for n = 2 and 3 were very high (85 and 53%) and surpassed even that of hydroxytamoxifen (38.5%), the active metabolite of tamoxifen. Ruthenocene derivatives act as anti-estrogens as effective (n = 2) or slightly more effective (n = 3-5) than hydroxytamoxifen on ERalpha-positive breast cancer cell lines but, unlike ferrocifens, do not show antiproliferative effects on ERalpha-negative breast cancer cell lines. Electrochemical studies showed that the ruthenocifen radical cations are unstable, which may account for this behavior. Some of these compounds could be useful as radiopharmaceuticals for ERalpha-positive breast cancer tumors. PMID- 15828820 TI - Synthesis and activity of substituted anthraquinones against a human filarial parasite, Brugia malayi. AB - Lymphatic filariasis (elephantiasis) is a global public health problem caused by the parasitic nematodes Wuchereria bancrofti and Brugia malayi. We have previously reported anthraquinones from daylily roots with potent activity against pathogenic trematode Schistosoma mansoni. Here we report the synthesis of novel anthraquinones A-S and their antifilrarial activity. Anthraquinones A-S were synthesized by a single-step Friedel-Crafts acylation reaction between phthalic anhydrides and substituted benzenes. The antifilarial properties of these synthetic anthraquinones were tested against microfilaria as well as adult male and female worms of B. malayi. The most active anthraquinone was K, which showed 100% mortality within 1, 5, and 3 days, respectively, against microfilaria and adult male and female worms at 5 ppm concentration. Albendazole, an oral drug currently used to treat parasitic infections, was used as a positive control. Methylated products of anthraquinones did not affect the microfilaria. Histological examination of treated adult female parasites showed most of the anthraquinones caused marked effects on intrauterine embryos. PMID- 15828821 TI - Redox-active antineoplastic ruthenium complexes with indazole: correlation of in vitro potency and reduction potential. AB - Antineoplastic ruthenium(III) complexes are generally regarded as prodrugs, being activated by reduction. Within a homologous series of ruthenium(III) complexes, cytotoxic potency is therefore expected to increase with increasing ease of reduction. Complexes of the general formula [Ru(III)Cl((6-n))(ind)n](3-n)- (n = 0 4; ind = indazole; counterions = Hind(+) or Cl(-)) and the compound trans [Ru(II)Cl(2)(ind)(4)] have been prepared and characterized electrochemically. Lever's parametrization method predicts that a higher indazole-to-chloride ratio results in a higher reduction potential, which is confirmed by cyclic voltammetry. In vitro antitumor potencies of these complexes in colon cancer cells (SW480) and ovarian cancer cells (CH1) vary by more than 2 orders of magnitude and increase in the following rank order: [Ru(III)Cl(6)](3-) < [Ru(III)Cl(4)(ind)(2)](-) < [Ru(III)Cl(5)(ind)](2-) << [Ru(III)Cl(3)(ind)(3)] < [Ru(III)Cl(2)(ind)(4)](+) approximately [Ru(II)Cl(2)(ind)(4)]. Thus, the observed differences in potency correlate with reduction potentials largely, though not perfectly, pointing to the influence of additional factors. Differences in the cellular uptake (probably resulting from different lipophilicity) contribute to this correlation but cannot solely account for it. PMID- 15828822 TI - Thrombospondin-1 mimetic peptide inhibitors of angiogenesis and tumor growth: design, synthesis, and optimization of pharmacokinetics and biological activities. AB - The heptapeptide 1, NAc-Gly-Val-DIle-Thr-Arg-Ile-ArgNHEt, a structurally modified fragment derived from the second type-1 repeat of thrombospondin-1 (TSP-1), is known to possess antiangiogenic activity. However, therapeutic utility could not be demonstrated because this peptide has a very short half-life in rodents. To optimize the PD/PK profile of 1, we initiated a systematic SAR study. The initial structural modifications were performed at positions 5 and 7 of peptide 1 and at the N- and C-termini. Out of several hundred peptides synthesized, the nonapeptide 5 (ABT-526) emerged as a promising lead. ABT-526 inhibited VEGF induced HMVEC cell migration and tube formation in the nanomolar range and increased apoptosis of HUAEC cells. ABT-526 showed acceptable PK in rodents, dog, and monkey. ABT-526, when incorporated in an angiogenic pellet implanted in the rat cornea at 10 microM, reduced neovascularization by 92%. Substitution of DalloIle in place of DIle in ABT-526 provided nonapeptide 6 (ABT-510), which was 30-fold less active than ABT-526 in the EC migration but 20-fold more active in the tube formation assay. In comparison to ABT-526, ABT-510 has increased water solubility and slower clearance in dog and monkey. Radiolabeled ABT-510 demonstrated saturable binding to HMVEC cells at 0.02-20 nM concentrations and was displaceable by TSP-1. ABT-510 and ABT-526 were shown to significantly increase apoptosis of HUAEC cells. ABT-510 was effective in blocking neovascularization in the mouse Matrigel plug model and inhibited tumor growth in the mouse Lewis lung carcinoma model. Previous studies had shown that ABT-510 was effective in inhibiting the outgrowth of murine melanoma metastases in syngeneic mice and in blocking the growth of human bladder carcinoma implanted in nude mice. It had been also shown that ABT-510 could regress tumor lesions in pet dogs or cause unexpected stabilization of the disease in advanced canine cancer. ABT 526 and ABT-510 are the first compounds in the class of potent inhibitors of angiogenesis that mimic the antiangiogenic function of TSP-1. ABT-510 is currently in phase II clinical studies. PMID- 15828823 TI - Studies on the mechanism of inactivation of the HIV-1 nucleocapsid protein NCp7 with 2-mercaptobenzamide thioesters. AB - The HIV-1 nucleocapsid protein (NCp7) is a small basic protein with two CysCysHisCys zinc-binding domains that specifically recognizes the Psi-site of the viral RNA. NCp7 plays a number of crucial roles in the viral lifecycle, including reverse transcription and RNA encapsidation. Several classes of potential anti-HIV compounds have been designed to inactivate NCp7 through zinc ejection, including a special class of thioester compounds. We have investigated the mechanism of action of two N-substituted-S-acyl-2-mercaptobenzamide compounds (compounds 1 and 2) that target NCp7. UV/Visible spectroscopy studies demonstrated that both thioesters were able to eject metal from NCp7. NMR and mass spectroscopy studies showed that the thioester compounds specifically ejected zinc from the carboxyl-terminal zinc-binding domain of NCp7 by covalent modification of Cys(39). Exposure of NCp7 to compounds 1 and 2 destroyed its ability to specifically bind RNA, whereas NCp7 already bound to RNA was protected from zinc ejection by the thioesters. The thiol component of the thioesters (compound 3, 2-mercaptobenzoyl-beta-alaninamide) did not eject zinc from NCp7, but when compound 3 was incubated with acetyl CoA prior to incubation with NCp7, we observed extensive metal ejection. Thus, the thiol released by the reaction of compounds 1 and 2 could be re-acylated in vivo by acyl CoA to form a new thioester compound that is able to react with NCp7. These studies provide a better understanding of the mechanism of action of thioester compounds, which is important for future design of anti-HIV-1 compounds that target NCp7. PMID- 15828824 TI - 1-Methyl-3H-pyrazolo[1,2-a]benzo[1,2,3,4]tetrazin-3-ones. Design, synthesis, and biological activity of new antitumor agents. AB - 1-Methylpyrazolo[1,2-a]benzo[1,2,3,4]tetrazin-3-ones 4, synthesized in good to excellent yields, were designed as novel alkylating agents because of their peculiar chemical behavior. All derivatives showed antiproliferative activity against more than 50 types of tumor cell lines with GI(50) reaching sub micromolar values. SAR studies revealed that the presence of a chlorine atom is well-tolerated in both positions 8 and 9, whereas in the case of the methyl group, switching from the 8 to the 9 position gives rise to the most active compound of the series, 4g, either for the number of cell lines inhibited and for selectivity against leukaemia and renal cancer subpanels. COMPARE and 3D-MIND computations indicate, for compounds 4, an activity profile analogous to rifamycins and cytidine analogues. PMID- 15828825 TI - Design, synthesis, and antiviral activity of adenosine 5'-phosphonate analogues as chain terminators against hepatitis C virus. AB - A series of adenosine 5'-phosphonate analogues were designed to mimic naturally occurring adenosine monophosphate. These compounds (1-5) were synthesized and evaluated in a cellular hepatitis C virus (HCV) replication assay. To improve cellular permeability and enhance the anti-HCV activity of these phosphonates, a bis(S-acyl-2-thioethyl) prodrug for compound 5 was prepared, and its cellular activity was determined. To elucidate the mechanism of action of these novel adenosine phosphonates, their diphosphate derivatives (1a-5a) were synthesized. Further nucleotide incorporation assays by HCV NS5B RNA-dependent RNA polymerase revealed that 2a and 3a can serve as chain terminators, whereas compounds 1a, 4a, and 5a are competitive inhibitors with ATP. Additional steady-state kinetic analysis determined the incorporation efficiency of 2a and 3a as well as the inhibition constants for 1a, 4a, and 5a. The structure-activity relationships among these compounds were analyzed, and the implication for nucleoside phosphonate drug design was discussed. PMID- 15828826 TI - Synthesis and pharmacology of ethylphenidate enantiomers: the human transesterification metabolite of methylphenidate and ethanol. AB - Ethanol elevates methylphenidate (1) plasma concentrations and yields the metabolite ethylphenidate (2). The therapeutic implications are under investigation. The IC(50) for dopamine reuptake inhibition by (+)-2 was 27 nM compared to 367 nM for cocaine and 1730 nM for (-)-2. Binding selectivity for dopamine versus norepinephrine transporters was greater for (+)-2 than for cocaine. Intraperitoneal (+)-2 was approximately half as active as (+)-1 in stimulating mouse motor activity at 5 mg/kg, but (+)-2 was as active as (+)-1 at 10 mg/kg. PMID- 15828827 TI - (5-Arylfuran-2-ylcarbonyl)guanidines as cardioprotectives through the inhibition of Na+/H+ exchanger isoform-1. AB - A series of (5-arylfuran-2-ylcarbonyl)guanidines was synthesized and evaluated for the NHE-1 inhibitory activity and cardiprotective efficacy against ischemia reperfusion injury. Starting with (5-phenylfuran-2-ylcarbonyl)guanidine 47 with a moderate inhibitory effect on NHE-1, the compounds with various substituents at the phenyl ring were investigated with the aim to optimize the potency. In this study, the 2,5-disubstituted compounds appeared to have better activities than the other analogues, and the 2-methoxy-5-chlorophenyl compound 85 was found as a potent inhibitor of NHE-1 (IC(50) = 0.081 microM). Furthermore, 85 showed a marked reduction of infarct size in the rat myocardial infarction model in vivo and significant improvement of cardiac contractile function in the isolated rat heart ischemia model in vitro. PMID- 15828828 TI - Evaluation of 8-arylsulfanyl, 8-arylsulfoxyl, and 8-arylsulfonyl adenine derivatives as inhibitors of the heat shock protein 90. AB - Hsp90 is a chaperone protein with important roles in maintaining transformation and in elevating the survival and growth potential of cancer cells. Currently there is an increasing interest in developing inhibitors of this protein as anticancer therapeutics. One of such inhibitors, the purine-scaffold class, has been reported to be potent and selective against Hsp90 both in vitro and in vivo models of cancer. Here, a series of 8-arylsulfanyl, -sulfoxyl, and -sulfonyl adenine members of the purine class was synthesized and evaluated as inhibitors of the chaperone. The structure-activity relationship and selectivity for tumor Hsp90 of compounds within the series is presented. Our results suggest that 8 arylsulfanyl adenine derivatives are good inhibitors of chaperone activity, whereas oxidation of the sulfides to sulfoxides or sulfones leads to compounds of decreased activity. The study identifies derivative 11v as the most potent Hsp90 inhibitor of the purine-scaffold series published to date (EC(50) = 30 nM), and also as the compound of this class with highest selectivity for tumor vs normal cell Hsp90 (700 to 3000-fold). Most rewardingly, this work has allowed for the identification of Hsp90 inhibitors with selective affinities for Hsp90-client protein complexes, derivatives that may represent useful pharmacological tools in dissecting Hsp90-regulated processes. PMID- 15828829 TI - Identification and characterization of novel benzil (diphenylethane-1,2-dione) analogues as inhibitors of mammalian carboxylesterases. AB - Carboxylesterases (CE) are ubiquitous enzymes responsible for the metabolism of xenobiotics. Because the structural and amino acid homology among esterases of different classes, the identification of selective inhibitors of these proteins has proved problematic. Using Telik's target-related affinity profiling (TRAP) technology, we have identified a class of compounds based on benzil (1,2 diphenylethane-1,2-dione) that are potent CE inhibitors, with K(i) values in the low nanomolar range. Benzil and 30 analogues demonstrated selective inhibition of CEs, with no inhibitory activity toward human acetylcholinesterase or butyrylcholinesterase. Analysis of structurally related compounds indicated that the ethane-1,2-dione moiety was essential for enzyme inhibition and that potency was dependent on the presence of, and substitution within, the benzene ring. 3D QSAR analyses of these benzil analogues for three different mammalian CEs demonstrated excellent correlations of observed versus predicted K(i) (r(2) > 0.91), with cross-validation coefficients (q(2)) of 0.9. Overall, these results suggest that selective inhibitors of CEs with potential for use in clinical applications can be designed. PMID- 15828830 TI - Structure-activity relationship studies optimizing the antiproliferative activity of novel cyclic somatostatin analogues containing a restrained cyclic beta-amino acid. AB - The cyclic somatostatin analogue cyclo[Pro(1)-Phe(2)-D-Trp(3)-Lys(4)-Thr(5) Phe(6)] (L-363,301) displays high biological activity in inhibiting the release of growth hormone, insulin, and glucagon. According to the sequence of L-363,301, we synthesized a number of cyclic hexa- and pentapeptides containing nonnatural alpha- and beta-amino acids. The N- fluorenylmethoxycarbonyl protected cyclic beta-amino acid [1S, 2S, 5R]-2-amino-3,5-dimethyl-2-cyclohex-3-enecarboxylic acid (cbetaAA), for the replacement of the Phe(6)-Pro(1) moiety of L-363,301, was synthesized in two steps by an enantioselective multicomponent reaction using (-) 8-phenylmenthol as a chiral auxiliary. The resulting peptide cyclo[cbetaAA(1) Tyr(2)-D-Trp(3)-Nle(4)-Thr(Trt)(5)] (Trt = triphenylmethyl) shows high antiproliferative effects in an in vitro assay with A431 cancer cells. The same peptide without the Trt group does not reveal any biological activity, whereas L 363,301 and closely related hexapeptides show only minor activity. By comparison of the solution structure of cyclo[cbetaAA(1)-Tyr(2)-D-Trp(3)-Nle(4)-Thr(Trt)(5)] with the structure of l-363,301, a nearly perfect match of the betaII'-turn region with d-Trp in the i + 1 position was observed. The cyclic beta-amino acid cbetaAA is likely needed for the bioactive conformation of the peptide. PMID- 15828831 TI - A pharmacophore hypothesis for P-glycoprotein substrate recognition using GRIND based 3D-QSAR. AB - Trying to understand the complex interactions that substrates and inhibitors have with the efflux transporter P-glycoprotein has been the subject of various publications. In this work, we have confined our study to substrates by picking a diverse set of 129 compounds based on the efflux ratios from Caco-2 permeability measurements. These compounds were then evaluated for P-glycoprotein inhibition using a calcein-AM assay. The subsequent data was used in a 3D-QSAR analysis using GRIND pharmacophore-based and physicochemical descriptors. Pharmacophore based descriptors produced a much more robust model than the one obtained from physicochemical-based descriptors. This supports the process proposed by Seelig and co-workers previously published whereby the substrate enters the membrane as the first step and is then recognized by P-glycoprotein in a second step. The strong correlation, highlighted by PLS statistical analysis, between pharmacophoric descriptors and inhibition values suggests that substrate interaction, with perhaps the mouth of the protein or another binding site, plays a key role in the efflux process, yielding a model in which diffusion across the membrane is less important than substrate-protein interaction. One pharmacophore emerged from the analysis of the model. We pose that the recognition elements, at least determined by the molecules used in this study, are two hydrophobic groups 16.5 A apart and two hydrogen-bond-acceptor groups 11.5 A apart and that the dimensions of the molecule also plays a role in its recognition as a substrate. PMID- 15828832 TI - High affinity central benzodiazepine receptor ligands: synthesis and biological evaluation of a series of phenyltriazolobenzotriazindione derivatives. AB - A series of 2-phenyl[1,2,3]triazolo[1,2-a][1,2,4]benzotriazin-1,5(6H)-diones (PTBTs), VII, were prepared and tested at the central benzodiazepine receptor (BzR). The skeleton of these compounds was designed by formally combining the N C=O moieties of the known BzR ligands, triazoloquinoxalines (IV) and triazinobenzimidazoles (ATBIs) (VI). Most of the PTBTs displayed submicromolar/nanomolar potency at the BzR. The 9-chloro derivatives (45-49) were generally found to be more potent than their 9-unsubstituted counterparts (37 44). Compound 45 turned out to be the most potent of the PTBTs (K(i) 2.8 nM). A subset of compounds (37, 42, 45, 49), when tested for their affinity on recombinant rat alpha1beta2gamma2, alpha2beta2gamma2, and alpha5beta3gamma2 GABA(A)/Bz receptor subtypes, showed enhanced affinities for the alpha1beta2gamma2 isoform, with compounds 45 and 49 exhibiting the highest selectivity. Moreover, compounds 45 and 49 were found to display a full agonist efficacy profile at alpha1 and alpha2 receptor subtypes, and an antagonist efficacy at alpha5-containing receptors. PMID- 15828833 TI - 3-Aminopyrazole inhibitors of CDK2/cyclin A as antitumor agents. 2. Lead optimization. AB - Inhibitors of cyclin-dependent kinases (CDK) such as CDK2/cyclin A-E are currently undergoing clinical trials to verify their potential as new anticancer agents. In a previous article we described the lead discovery process of a 3 aminopyrazole class of CDK2/cyclin A-E inhibitors. The endpoint of this process was PNU-292137, a compound endowed with in vivo antitumor activity in a mouse tumor xenograft model. We optimized this lead compound to improve some physicochemical properties, notably solubility and plasma protein binding. This lead optimization process brought us to the discovery of (2S)-N-(5-cyclopropyl-1H pyrazol-3-yl)-2-[4-(2-oxo-1-pyrrolidinyl)phenyl]propanamide (PHA-533533, 13), a compound with a balanced activity vs druglike profile. Compound 13 inhibited CDK2/cyclin A with a K(i) of 31 nM, counteracting tumor cell proliferation of different cell lines with an IC(50) in the submicromolar range. Solubility was improved more than 10 times over the starting lead, while plasma protein binding was decreased from 99% to 74%. With exploitation of this globally enhanced in vitro profile, 13 was more active than PNU-292137 in vivo in the A2780 xenograft model showing a tumor growth inhibition of 70%. Proof of mechanism of action was obtained in vivo by immunohistochemical analysis of tumor slices of 13-treated vs untreated animals. PMID- 15828834 TI - Pyridinium-1-yl bisphosphonates are potent inhibitors of farnesyl diphosphate synthase and bone resorption. AB - We report the design, synthesis and testing of a series of novel bisphosphonates, pyridinium-1-yl-hydroxy-bisphosphonates, based on the results of comparative molecular similarity indices analysis and pharmacophore modeling studies of farnesyl diphosphate synthase (FPPS) inhibition, human Vgamma2Vdelta2 T cell activation and bone resorption inhibition. The most potent molecules have high activity against an expressed FPPS from Leishmania major, in Dictyostelium discoideum growth inhibition, in gammadelta T cell activation and in an in vitro bone resorption assay. As such, they represent useful new leads for the discovery of new bone resorption, antiinfective and anticancer drugs. PMID- 15828835 TI - Dimeric zanamivir conjugates with various linking groups are potent, long-lasting inhibitors of influenza neuraminidase including H5N1 avian influenza. AB - The synthesis, antiviral and pharmacokinetic properties of zanamivir (ZMV) dimers 8 and 13 are described. The compounds are highly potent neuraminidase (NA) inhibitors which, along with dimer 3, are being investigated as potential second generation inhaled therapies both for the treatment of influenza and for prophylactic use. They show outstanding activity in a 1 week mouse influenza prophylaxis assay, and compared with ZMV, high concentrations of 8 and 13 are found in rat lung tissue after 1 week. Retention of compounds in rat lung tissue correlated both with molecular weight (excluding 3 and 15) and with a capacity factor K' derived from immobilized artificial membrane (IAM) chromatography (including 3 and 15). Pharmacokinetic parameters for 3, 8 and 13 in rats show the compounds have short to moderate plasma half-lives, low clearances and low volumes of distribution. Dimer 3 shows NA inhibitory activity against N1 viruses including the recent highly pathogenic H5N1 A/Chicken/Vietnam/8/2004. In plaque reduction assays, 3, 8 and 13 show good to outstanding potency against a panel of nine flu A and B virus strains. Consistent with its shorter and more rigid linking group, dimer 8 has been successfully crystallized. PMID- 15828836 TI - Novel C-17-heteroaryl steroidal CYP17 inhibitors/antiandrogens: synthesis, in vitro biological activity, pharmacokinetics, and antitumor activity in the LAPC4 human prostate cancer xenograft model. AB - New chemical entities, steroidal C-17 benzoazoles (5, 6, 9 and 10) and pyrazines (14 and 15) were rationally designed and synthesized. The key reaction for synthesis of the benzoazoles involved the nucleophilic vinylic "addition elimination" substitution reaction of 3beta-acetoxy-17-chloro-16-formylandrosta 5,16-diene (2) and benzoazole nucleophiles, while that for synthesis of pyrazines involved palladium-catalyzed cross-coupling reaction of 17-iodoandrosta-5,16-dien 3beta-ol (13) with tributylstannyl diazines. Some of the compounds were shown to be potent inhibitors of human CYP17 enzyme as well as potent antagonist of both wild type and mutant androgen receptors (AR). The most potent CYP17 inhibitors were 3beta-hydroxy-17-(1H-benzimidazole-1-yl)androsta-5,16-diene (5, code named VN/124-1), 3beta-hydroxy-17-(5(1)-pyrimidyl)androsta-5,16-diene (15) and 17-(1H benzimidazole-1-yl)androsta-4,16-dien-3-one (6), with IC(50) values of 300, 500 and 915 nM, respectively. Compounds 5, 6, 14 and 15 were effective at preventing binding of (3)H-R1881 (methyltrienolone, a stable synthetic androgen) to both the mutant LNCaP AR and the wild-type AR, but with a 2.2- to 5-fold higher binding efficiency to the latter. Compounds 5 and 6 were also shown to be potent pure AR antagonists. The cell growth studies showed that 5 and 6 inhibit the growth of DHT-stimulated LNCaP and LAPC4 prostate cancer cells with IC(50) values in the low micromolar range (i.e., <10 microM). Their inhibitory potencies were comparable to that of casodex but remarkably superior to that of flutamide. The pharmacokinetics of compounds 5 and 6 in mice were investigated. Following s.c. administration of 50 mg/kg of 5 and 6, peak plasma levels of 16.82 and 5.15 ng/mL, respectively, occurred after 30 to 60 min, both compounds were cleared rapidly from plasma (terminal half-lives of 44.17 and 39.93 min, respectively), and neither was detectable at 8 h. Remarkably, compound 5 was rapidly converted into a metabolite tentatively identified as 17-(1H-benzimidazol-1-yl)androsta-3 one. When tested in vivo, 5 proved to be very effective at inhibiting the growth of androgen-dependent LAPC4 human prostate tumor xenograft, while 6 was ineffective. Compound 5 (50 mg/kg/twice daily) resulted in a 93.8% reduction (P = 0.00065) in the mean final tumor volume compared with controls, and it was also significantly more effective than castration. To our knowledge, this is the first example of an antihormonal agent (an inhibitor of androgen synthesis (CYP17 inhibitor)/antiandrogen) that is significantly more effective than castration in suppression of androgen-dependent prostate tumor growth. In view of these impressive anticancer properties, compound 5 is a strong candidate for development for the treatment of human prostate cancer. PMID- 15828837 TI - Imaging of melanoma using 64Cu- and 86Y-DOTA-ReCCMSH(Arg11), a cyclized peptide analogue of alpha-MSH. AB - Early detection of melanoma is essential, since a patient's prognosis with metastatic melanoma is poor. Previous studies showed that (111)In-DOTA ReCCMSH(Arg(11)), a cyclic analogue of alpha-melanocyte stimulating hormone (alpha-MSH), exhibited high tumor concentration and rapid clearance from nontarget tissue. The goal of this current study was to label DOTA ReCCMSH(Arg(11)) with beta(+)-emitting radionuclides, to determine if the high sensitivity of positron emission tomography (PET) imaging would aid in the detection of malignant melanoma. DOTA-ReCCMSH(Arg(11)) was labeled with (64)Cu and (86)Y. Biodistribution and small animal PET imaging were carried out in mice implanted with B16/F1 murine melanoma tumor and compared with data obtained in the same animal model with [(18)F]FDG. In both cases a subset of animals were co injected with 20 microg of DOTA-ReCCMSH(Arg(11)) to determine if tumor concentration was receptor mediated. Tumor concentration for both the (86)Y- and (64)Cu-complexes reached a maximum at 30 min, while coadministering 20 microg of unlabeled complex reduced tumor uptake significantly. Nontarget organ concentration was considerably lower with (86)Y-DOTA-ReCCMSH(Arg(11)) than its (64)Cu analogue, except in the kidneys, where the (64)Cu complex had lower accumulation at all time points. Small animal PET images for both complexes showed the tumor could be visualized after 30 min, with the standardized uptake value (SUV) analysis following a similar trend as the biodistribution data. The data obtained suggests that DOTA-ReCCMSH(Arg(11)), when labeled with beta(+) emitting radionuclides, has the potential for early detection of malignant melanoma by exploiting the sensitivity and high resolution of PET. PMID- 15828838 TI - Conformationally restricted analogues of psorospermin: design, synthesis, and bioactivity of natural-product-related bisfuranoxanthones. AB - The antileukemic xanthone psorospermin is a topoisomerase II-dependent DNA alkylator in advanced preclinical development. Efforts have been made to further understand the structural requirements of its mechanism of action through the synthesis of ring-constrained analogues, based on the skeleton of the bisfuranoxanthone natural products. Molecules were designed that contain the bisfuran and xanthone portions of naturally occurring psorofebrins, and molecular modeling was used to assess their DNA alkylating potential and to refine the structures. A short, diastereoselective synthetic process to access bisfuranoxanthones was developed, culminating in the first total synthesis of (+/ )-isohydroxypsorofebrin. Two compounds designed and synthesized were of particular interest, chlorohydrin 7 and epoxide 6, which are reactive analogues of the natural product isohydroxypsorofebrin. The chlorohydrin retains the psorospermin-like DNA alkylation characteristics despite its rigid structure and high innate affinity for DNA. Molecular modeling has been used to rationalize the increased activity of the chlorohydrin. The chlorohydrin and epoxide show increased cytotoxicity compared to isohydroxypsorofebrin against a range of human tumor cell lines. PMID- 15828839 TI - Applying linear interaction energy method for rational design of noncompetitive allosteric inhibitors of the sarco- and endoplasmic reticulum calcium-ATPase. AB - Noncompetitive inhibitors of sarco- and endoplasmic reticulum calcium-ATPase (SERCA) have important therapeutic value in the treatment of cancer, due to their ability to induce apoptosis in cancer cells in a proliferation-independent manner. Thapsigargin (TG) and its analogues are one such class of inhibitors that bind to a hydrophobic pocket located in the transmembrane region of SERCA near the biomembrane surface and interfere with calcium transport. The binding free energies of thapsigargin-based inhibitors of SERCA were computed using a novel linear interaction energy (LIE) method with a surface generalized Born (SGB) continuum solvation model. A training set of 20 TG analogues was used to build a binding affinity model for estimating the free energy of binding for 18 new inhibitors with a root-mean-square (rms) error of 1.36 kcal/mol with respect to experimental data. For 15 out of the 18 inhibitors in the test set, the rms error was 1.02 kcal/mol, which is on the order of the accuracy level achieved by highly rigorous free energy of perturbation (FEP) or thermodynamic integration (TI) methods. On the basis of the analysis of the binding cavity at the interface of the membrane surface and the cytoplasmic region, we propose that side chains of TG derivatives at the O-8 position orient toward the cytoplasmic region through a hydrophobic channel. On the basis of this insight, four analogues of varying side chain length at the O-8 position with a charged moiety at the end were designed, tested with LIE methodology, and then validated experimentally for their SERCA inhibition activity. Low levels of rms error for the majority of inhibitors establish the structure-based LIE method as an efficient tool for generating more potent and specific inhibitors of SERCA by testing rationally designed lead compounds based on thapsigargin derivatization. PMID- 15828840 TI - Additivity of molecular fields: CoMFA study on dual activators of PPARalpha and PPARgamma. AB - Recent trends in drug discovery include methods to identify dual and triple activating drugs. This approach is being successfully employed in malaria, cancer, asthma, insulin resistance, etc. Molecular field analysis has been employed in correlating pharmacological data and field parameters. In this paper we introduce the concept of additivity of molecular fields to correlate molecular fields of dual activators and their pIC(50) values. PPARalpha and PPARgamma dual activators, which affect hypertriglyceridemia and hyperglycemia, have been chosen to validate the molecular field additivity concept. Three CoMFA models namely alpha-model, gamma-model and dual-model have been developed. The validity of this concept has been ascertained by (a) comparing contour maps, (b) by comparing CoMFA results with FlexX docking results and (c) by analyzing newly designed molecules. PMID- 15828842 TI - Differential inhibition of HIV-1 and SIV envelope-mediated cell fusion by C34 peptides derived from the C-terminal heptad repeat of gp41 from diverse strains of HIV-1, HIV-2, and SIV. AB - The spectrum of inhibition of human (HIV) and simian (SIV) immunodeficiency virus envelope (Env)-mediated cell fusion by C34, a 34 residue peptide corresponding to the C-heptad repeat of gp41 (residues 628-661 of HIV-1 Env), has been examined using a panel of five envelope glycoproteins, three from HIV-1 (LAV, SF162 and 89.6) and two from SIV (mac239 and mac316), and six C34 peptides derived from three strains of HIV-1 (LAV, N CM, and O CM), two strains of HIV-2 (EHO and ALI), and one strain of SIV (African Green Monkey, AGM). A quantitative vaccinia-based reporter gene cell fusion assay was employed. The inhibition data from the panel of 30 C34/envelope glycoprotein combinations, which can be fit to a simple activity relationship with IC(50) values spanning a range of over 4 orders of magnitude from 4 nM to 70 microM, permits one to rationalize both the potency and broadness of the inhibitory properties of the C34 peptides in terms of computed interaction free energies between the C34 peptides and the N-helical trimeric coiled-coil of gp41 and the helical propensities of the free C34 peptides. Of particular interest is the finding that the C34 peptide derived from the EHO strain of HIV-2 is a broad spectrum, highly potent inhibitor of Env-mediated cell fusion with IC(50) values spanning a very narrow range from only 4 to 25 nM over the entire panel of HIV-1 and SIV envelope glycoproteins tested. This result suggests that C34 from HIV-2 EHO may present a potentially useful therapeutic agent against diverse and/or resistant strains of HIV-1. PMID- 15828841 TI - Novel cyclopropyl beta-amino acid analogues of pregabalin and gabapentin that target the alpha2-delta protein. AB - As part of a program aimed at generating compounds with affinity for the alpha(2) delta subunit of voltage-gated calcium channels, several novel beta-amino acids were prepared using an efficient nitroalkane-mediated cyclopropanation as a key step. Depending on the ester that was chosen, the target amino acids could be prepared in as few as three steps. The cyclopropyl amino acids derived from ketones proved to be potent binders of the alpha(2)-delta subunit of voltage gated calcium channels, but did not interact with the large neutral amino acid system L (leucine) transporter. Anticonvulsant effects were observed in vivo with compound 34 but only after intracerebroventricular (icv) administration, presumably due to inadequate brain concentrations of the drug being achieved following oral dosing. However, pregabalin 1 was active in the DBA/2 model after oral (and icv) dosing, supporting a hypothesis that active transport is a prerequisite for such zwitterionic species to cross the blood-brain barrier. PMID- 15828843 TI - Combinatorial optimization of isatin-beta-thiosemicarbazones as anti-poxvirus agents. AB - Novel strategies are required to combat pox virus infections, whether caused by escape of viruses such as monkeypox from indigenous areas or intentional release of smallpox. Anti-smallpox drugs with a unique mode of antiviral action, inhibition of transcription termination, were known but not therapeutically useful. Using a combinatorial method, variants of the basic isatin-beta thiosemicarbazone structure were prepared and examined for cytotoxicity and antiviral activity in vaccinia virus- and cowpox virus-infected human cells. Potent and much more selective N-aminomethyl-isatin-beta-thiosemicarbazones were discovered. PMID- 15828844 TI - Neurosteroid analogues. 10. The effect of methyl group substitution at the C-6 and C-7 positions on the GABA modulatory and anesthetic actions of (3alpha,5alpha)- and (3alpha,5beta)-3-hydroxypregnan-20-one. AB - The planar 5alpha-reduced steroid (3alpha,5alpha)-3-hydroxypregnan-20-one and the nonplanar 5beta-reduced steroid (3alpha,5beta)-3-hydroxypregnan-20-one act at GABA(A) receptors to induce general anesthesia. The structural features of the binding sites for these anesthetic steroids on GABA(A) receptors have not been determined. To determine how structural modifications at the steroid C-6 and C-7 positions effect the actions of these anesthetic steroids, an axial or equatorial methyl group was introduced at these positions. The analogues were evaluated (1) in [(35)S]-tert-butylbicyclophosphorothionate binding experiments, (2) in electrophysiological experiments using rat alpha(1)beta(2)gamma(2L) GABA(A) receptors expressed in Xenopus laevis oocytes, and (3) as tadpole anesthetics. The effects of methyl group substitution in the 5alpha- and 5beta-reduced series of compounds were strikingly similar. In both series, a 6beta-Me group gave compounds with actions similar to or greater than those of the parent steroids. A 6alpha-, 7beta- or 7alpha-Me substituent resulted in reduced potency for inhibition of radioligand binding, GABA(A) receptor modulation and tadpole anesthesia. Because of the similar effects of methyl group substitution in the two series of compounds and previous results from other studies showing that structural modifications in the steroid D ring/side chain region produce similar effects regardless of the stereochemistry of the A,B-ring fusion, we propose that either the 3alpha-hydroxyl groups of planar and nonplanar anesthetic steroids hydrogen bond to different amino acids on GABA(A) receptors or that this critical hydrogen bonding group interacts with membrane lipids instead of the receptor. PMID- 15828845 TI - Structure-activity relationships of the unique and potent agouti-related protein (AGRP)-melanocortin chimeric Tyr-c[beta-Asp-His-DPhe-Arg-Trp-Asn-Ala-Phe-Dpr]-Tyr NH2 peptide template. AB - The melanocortin receptor system consists of endogenous agonists, antagonists, G protein coupled receptors, and auxiliary proteins that are involved in the regulation of complex physiological functions such as energy and weight homeostasis, feeding behavior, inflammation, sexual function, pigmentation, and exocrine gland function. Herein, we report the structure-activity relationship (SAR) of a new chimeric hAGRP-melanocortin agonist peptide template Tyr-c[beta Asp-His-DPhe-Arg-Trp-Asn-Ala-Phe-Dpr]-Tyr-NH(2) that was characterized using amino acids previously reported in other melanocortin agonist templates. Twenty peptides were examined in this study, and six peptides were selected for (1)H NMR and computer-assisted molecular modeling structural analysis. The most notable results include the identification that modification of the chimeric template at the His position with Pro and Phe resulted in ligands that were nM mouse melanocortin-3 receptor (mMC3R) antagonists and nM mouse melanocortin-4 receptor (mMC4R) agonists. The peptides Tyr-c[beta-Asp-His-DPhe-Ala-Trp-Asn-Ala-Phe-Dpr] Tyr-NH(2) and Tyr-c[beta-Asp-His-DNal(1')-Arg-Trp-Asn-Ala-Phe-Dpr]-Tyr-NH(2) resulted in 730- and 560-fold, respectively, mMC4R versus mMC3R selective agonists that also possessed nM agonist potency at the mMC1R and mMC5R. Structural studies identified a reverse turn occurring in the His-DPhe-Arg-Trp domain, with subtle differences observed that may account for the differences in melanocortin receptor pharmacology. Specifically, a gamma-turn secondary structure involving the DPhe(4) in the central position of the Tyr-c[beta-Asp-Phe DPhe-Arg-Trp-Asn-Ala-Phe-Dpr]-Tyr-NH(2) peptide may differentiate the mixed mMC3R antagonist and mMC4R agonist pharmacology. PMID- 15828846 TI - Synthesis and structure-activity relationship of fluoro analogues of 8-{2-[4-(4 methoxyphenyl)piperazin-1yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione as selective alpha(1d)-adrenergic receptor antagonists. AB - We have discovered high-affinity antagonists (exemplified by 11 and 12) that are the most selective for alpha(1d)-adrenergic receptors (alpha(1d)-AR) reported to date. In cloned receptor assay systems, 12 displays at least 95-fold selectivity for the alpha(1d)-AR over all other G-protein-coupled receptors tested, and the subtype selectivity of 11 was confirmed in pharmacologically defined isolated tissue preparations. PMID- 15828847 TI - Potent and selective Aurora inhibitors identified by the expansion of a novel scaffold for protein kinase inhibition. AB - Potent and selective Aurora kinase inhibitors were identified from the combinatorial expansion of the 1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole bi-cycle, a novel and versatile scaffold designed to target the ATP pocket of protein kinases. The most potent compound reported in this study had an IC(50) of 0.027 microM in the enzymatic assay for Aur-A inhibition and IC(50)s between 0.05 microM and 0.5 microM for the inhibition of proliferation of different tumor cell lines. PMID- 15828848 TI - Potential antitumor agents. 37. Synthesis and antitumor activity of guanylhydrazones from imidazo[2,1-b]thiazoles and from the new heterocyclic system thiazolo[2',3':2,3]imidazo[4,5-c]quinoline. AB - This paper reports synthesis and antitumor activity of new guanylhydrazones from imidazo[2,1-b]thiazoles and from the new heterocyclic system thiazolo[2',3':2,3]imidazo[4,5-c]quinoline. The compounds were tested as potential antitumor agents at the National Cancer Institute. The effect of the guanylhydrazone of 2-chloro-6-(2,5-dimethoxy-4-nitrophenyl)imidazo[2,1-b]thiazole 5-carbaldehyde (41) was investigated, and it was found to be an inhibitor of Complex III of the mitochondrial respiratory chain and is able to induce apoptosis in the cell lines HT29 and HL60. PMID- 15828849 TI - Pharmacokinetic and pharmacodynamic considerations in the development of therapeutic proteins. AB - With an increasing number of therapeutic proteins moving into preclinical and clinical development, pharmacokinetic factors play an important role in the development of these macromolecules. It is also important that the pharmacokinetic evaluation of these compounds be done as accurately as possible. For macromolecules, evaluation of pharmacokinetic parameters is often complicated by a number of factors. Bioanalytical methods are essential for any pharmacokinetic study, but for many therapeutic proteins the immunoassay and bioassay methodologies are often nonspecific and sometimes the estimation of pharmacokinetic parameters becomes assay dependent. In vivo binding proteins, metabolites and antibody formation may also interfere with bioanalytical methodologies and thus may have significant impact on the pharmacokinetics of therapeutic proteins. There are also difficulties in identifying and quantifying metabolites as well as the binding of therapeutic proteins to endogenous proteins. Some macromolecules exhibit species specificity that complicates the preclinical pharmacological and toxicological evaluation of these compounds. Antibody formation is a particular problem in the preclinical evaluation of therapeutic proteins. Changes in structure or sequence of protein molecules (glycosylation or pegylation) may cause changes in the pharmacokinetics of these compounds. The size of therapeutic proteins may become a hindrance for absorption. Low absorption of intact molecules across biological membranes frequently occurs. Other factors that may affect the pharmacokinetics of a therapeutic protein are immunogenicity, presence of endogenous protein, time of drug administration, and rate and site of drug delivery. The relationship between pharmacokinetics and pharmacodynamics of therapeutic proteins is complex and in most cases is unclear. In many cases the mechanism and site of action are unknown for these compounds. PMID- 15828851 TI - Pharmacokinetic/pharmacodynamic relationships of asparaginase formulations: the past, the present and recommendations for the future. AB - The discovery of the tumour-inhibitory properties of asparaginase began 50 years ago with the observation that guinea-pig serum-treated lymphoma-bearing mice underwent rapid and often complete regression. Soon afterwards, the asparaginase of bacterial origin was isolated. The asparaginases of bacterial origin induce anti-asparaginase neutralising antibodies in a large proportion of patients (44 60%), thus negating the specific enzymatic activity and resulting in failure of the target amino acid deamination in serum. There is immunological cross-reaction between the antibodies against various formulations of native Escherichia coli asparaginase and polyethylene glycol (PEG)-asparaginases, but not to Erwinia asparaginase, as suggested by laboratory preclinical findings. This evidence was strongly inferred from the interim analyses in the Children's Cancer Group (CCG) 1961 study. Thus, anti-E. coli or PEG-asparaginase antibodies seropositive patients may benefit from the Erwinia asparaginase. The inter-relationships between asparaginase activity, asparagine (ASN) and glutamine deamination remain largely unexplored in patients. Studies have shown that ASN depletion is insufficient to induce apoptosis in T lymphoblasts in vitro and that the inhibitory concentration of CEM T-cell line is correlated with the asparaginase concentration responsible for 50% glutamine deamination. The optimal catalysis of ASN and glutamine deamination in serum by asparaginase induces apoptosis of leukaemic lymphoblasts. The percentage of ASN and glutamine deamination was predicted by asparaginase activity. Asparaginase activity of 0.1 IU/mL provided insufficient depletion of both amino acids in high-risk acute lymphoblastic leukaemia (ALL) patients. With increasing glutamine deamination, mean asparaginase activities and percentages of post-treatment samples with effective ASN depletion (<3 micromol/L) increase. Both glutamine and ASN deamination are predicted by asparaginase activity. Further population analyses resulted in identification of sigmoid relationships between asparaginase levels and post treatment glutamine and ASN deamination.Furthermore, pharmacodynamic analyses strongly suggested that >/=90% deamination of glutamine must occur before optimal ASN deamination takes place, due to the de novo ASN biosynthesis by the liver. These pharmacodynamic results from the best-fit population pharmacokinetic/pharmacodynamic model obtained from nonlinear mixed effects model pharmacodynamic analyses for standard-risk ALL patients are similar. These analyses produced the following results: (i) asparaginase activity 0.4-0.7 IU/mL was required for optimal (90%) ASN and glutamine deamination; and (ii) deamination of glutamine is dependent on asparaginase activity and it correlates with enhanced serum ASN deamination. Thus, glutamine deamination enhances asparaginase efficacy in ALL patients. Deamination of ASN >/=90% of control or ASN concentration <3 micromol/L may be associated with improved survival in this subset of patients. Our findings support the pharmacodynamic mechanism of PEG-asparaginase for disease control in ALL patients. These results taken together strongly support new experimental approaches for application of population pharmacokinetic/pharmacodynamic analyses to further enhance survival of leukaemia patients. PMID- 15828852 TI - Pharmacokinetics of famotidine in infants. AB - BACKGROUND: Although famotidine pharmacokinetics are similar in adults and children older than 1 year of age, they differ in neonates owing to developmental immaturity in renal function. Little is currently known about the pharmacokinetics of famotidine in infants aged between 1 month and 1 year, a period when renal function is maturing. OBJECTIVE: To characterise the pharmacokinetics of famotidine in infants. DESIGN: This was a two-part multicentre study with both single dose (Part I, open-label) and multiple dose (Part II, randomised) arms. PATIENTS: Thirty-six infants (20 females and 16 males) who required treatment with famotidine and who had an indwelling arterial or venous catheter for reasons unrelated to the study. METHODS: Infants in Part I were administered a single dose of famotidine 0.5 mg/kg; the dose was intravenous or oral according to the judgement of the attending physician. Infants receiving 0.5 mg/kg intravenously were divided into two groups by age, and pharmacokinetic parameters in infants 0-3 months and >3 to 12 months of age were compared. Infants in Part II were randomised to one of the following treatments: 0.25 mg/kg/dose intravenously or 0.5 mg/kg/dose orally on day 1 and subsequent days, or 0.25 mg/kg/dose intravenously or 0.5 mg/kg/dose orally on day 1 followed by doses of either 0.5 mg/kg/dose intravenously or 1 mg/kg/dose orally on subsequent days. From day 2 onwards, age-adjusted dose administration regimens (once daily in infants <3 months of age and every 12 hours in infants >3 months of age) were used; the total number of famotidine doses ranged from 3 to 11 and the total number of days of dose administration ranged from two to eight. RESULTS: In infants <3 months of age, plasma and renal clearance of famotidine were decreased compared with infants >3 months of age. Pharmacokinetic parameters for the older infants (i.e. those >3 months) were similar to those previously reported for children and adults. Approximate dose-proportionality, no accumulation on multiple dosing and an estimated bioavailability similar to adult values were also observed. CONCLUSION: A short course of famotidine therapy in infants appears generally well tolerated, and the characteristics of famotidine pharmacokinetics during the first year of life are explained to a great degree by the development of renal function, the primary route of elimination for this drug. PMID- 15828850 TI - Role of cytochrome P450 activity in the fate of anticancer agents and in drug resistance: focus on tamoxifen, paclitaxel and imatinib metabolism. AB - Although activity of cytochrome P450 isoenzymes (CYPs) plays a major role in the fate of anticancer agents in patients, there are relatively few clinical studies that evaluate drug metabolism with therapeutic outcome. Nevertheless, many clinical reports in various non-oncology fields have shown the dramatic importance of CYP activity in therapeutic efficacy, safety and interindividual variability of drug pharmacokinetics. Moreover, variability of drug metabolism in the liver as well as in cancer cells must also be considered as a potential factor mediating cancer resistance. This review underlines the role of drug metabolism mediated by CYPs in pharmacokinetic variability, drug resistance and safety. As examples, biotransformation pathways of tamoxifen, paclitaxel and imatinib are reviewed. This review emphasises the key role of therapeutic drug monitoring as a complementary tool of investigation to in vitro data. For instance, pharmacokinetic data of anticancer agents have not often been published within subpopulations of patients who show ultra-rapid, extensive or poor metabolism (e.g. due to CYP2D6 and CYP2C19 genotypes). Besides kinetic variability in the systemic circulation, induction of CYP activity may participate in creating drug resistance by speeding up the cancer agent degradation specifically in the target cells. For one cancer agent, various mechanisms of resistance are usually identified within different cell clones. This review also tries to emphasise that drug resistance mediated by CYP activity in cancer cells should be taken into consideration to a greater degree. The unequivocal identification of the metabolising enzymes involved in clinical conditions will eventually allow improvement and individualisation of anticancer agent therapy, i.e. drug dosage and selection. In addition, a more complete understanding of the metabolism of anticancer agents will assist in the prediction of drug-drug interactions, as anticancer agent combinations are becoming more prevalent. PMID- 15828853 TI - Topiramate pharmacokinetics in children and adults with epilepsy: a case-matched comparison based on therapeutic drug monitoring data. AB - OBJECTIVE: To compare the steady-state pharmacokinetics of topiramate in a large population of children and adults with epilepsy in a therapeutic drug monitoring setting. STUDY DESIGN: Retrospective, case-matched pharmacokinetic evaluation. PATIENTS: Seventy children (aged 1-17 years) with epilepsy and 70 adult controls (aged 18-65 years) with epilepsy, matched for sex and comedication. METHODS: Topiramate apparent oral clearance (CL/F) values were calculated from steady state serum concentrations in children and compared with those determined in controls. Comparisons were made by means of the Mann-Whitney's U-test, or the Kruskal-Wallis test in the case of multiple comparisons. A linear regression model was used to assess potential correlation of CL/F values with age. To investigate the influence of different variables on the variability in topiramate CL/F values, a multiple regression model was developed. RESULTS: In the absence of enzyme-inducing comedication, mean topiramate CL/F was 42% higher in children than in adults (40.3 +/- 21.0 vs 28.4 +/- 15.3 mL/h/kg; p < 0.01). In children and adults comedicated with enzyme-inducing antiepileptic drugs (AEDs), topiramate CL/F values were approximately 1.5- to 2-fold higher than those observed in the absence of enzyme inducers, and the elevation in topiramate CL/F in children compared with adults was also present in the subgroups receiving enzyme inducers (66%; 76.6 +/- 35.1 vs 46.1 +/- 16.7 mL/h/kg; p < 0.0001). In the paediatric population, a negative correlation between CL/F and age was demonstrated, both in the absence (p < 0.01) and in the presence (p < 0.001) of enzyme induction. The independent influence of age and enzyme-inducing AEDs on topiramate CL/F was confirmed by multiple regression analysis. CONCLUSION: Topiramate CL/F is highest in young children and decreases progressively with age until puberty, presumably due to age-dependent changes in the rate of drug metabolism. As a result of this, younger patients require higher dosages to achieve serum topiramate concentrations comparable with those found in older children and adults. Enzyme-inducing comedication decreases serum topiramate concentration by approximately one-half and one-third in children and adults, respectively. PMID- 15828854 TI - Pharmacokinetic/pharmacodynamic and time-to-event models of ribavirin-induced anaemia in chronic hepatitis C. AB - BACKGROUND: Interferon (IFN)-alpha and ribavirin combination therapy is the standard treatment for patients with chronic hepatitis C. However, ribavirin induces anaemia, especially by haemolysis, an adverse effect that is dose limiting. OBJECTIVES: The aim of this study was to determine the relationships between ribavirin exposure and haemoglobin time-course, the time-to-anaemia and the covariates influencing these relationships in a population of patients treated for chronic hepatitis C. In addition, we also intended to establish a simple rule defining the need for dosage adjustment, using data obtained during the first month of treatment. METHODS: A retrospective analysis of 99 patients treated with IFNalpha plus ribavirin, with known dosage administration history, liver biopsy, demographic data, red blood cell counts, haemoglobin level (1037 measurements, median 10 per patient, range 2-31) and serum creatinine during the entire treatment period (178 days, range 53-382 days) was conducted. The data were analysed by a pharmacokinetic/pharmacodynamic population model and Weibull time-to-anaemia model. The rule defining the need for dosage adjustment was as follows: adjustment was needed if haemoglobin at steady state (H(ss)), estimated by the Bayesian method based on data obtained during the first month of treatment, was <12 g/dL for men or <11 g/dL for women. RESULTS: In both models, anaemia was related to the exposure of erythrocytes to ribavirin at time t (RT in mg/kg/day) by a maximum effect model, with RT(50) (dosage administration rate at which half the maximal effect is reached) approximately 12 mg/kg/day, and the significant covariates were initial haemoglobin level and bodyweight. Performances of a Bayesian prediction of H(ss) based on two early haemoglobin level measurements were encouraging (mean prediction error 0.12 g/dL, precision 0.85 g/dL). The proposed rule for the need of dosage adjustment was able to predict the actual evolution of the dosage regimen in 76% of non-adapted patients and 69% of adapted patients. CONCLUSION: The current guidelines for ribavirin dosage administration, based on bodyweight, are adequate, at least in the 45-105 kg range. Results indicate that Bayesian therapeutic monitoring could be helpful in controlling ribavirin-induced anaemia. PMID- 15828855 TI - Absence of pharmacokinetic interactions of the combined contraceptive vaginal ring NuvaRing with oral amoxicillin or doxycycline in two randomised trials. AB - BACKGROUND: Two pharmacokinetic studies were performed to investigate whether there is any interaction between etonogestrel or ethinylestradiol released from the combined contraceptive vaginal ring NuvaRing and concomitant treatment with orally administered amoxicillin or doxycycline. METHODS: In one study, healthy women were randomised to receive either NuvaRing alone for 21 days or NuvaRing for 21 days plus amoxicillin on days 1-10. After a 7-day ring-free washout period, women were crossed over to the alternate regimen for a further 21-day treatment period. The other study used an identical design except that women received doxycycline instead of amoxicillin. The amoxicillin study measured serum etonogestrel and ethinylestradiol levels and area under the serum concentration time curve (AUC) values over the initial 12 hours on days 1 (AUC(12)) and 10 (AUC(216-228)) and the whole of days 1-11 (AUC(240)) and 1-22 (AUC(504)). The doxycycline study measured AUC values over the initial 24 hours on days 1 (AUC(24)) and 10 (AUC(216-240)) and the whole of days 1-11 (AUC(240)) and 1-22 (AUC(504)). RESULTS: No differences in etonogestrel or ethinylestradiol serum concentrations were observed between subjects using NuvaRing alone versus those receiving the ring plus either of the antibiotics. Calculation of etonogestrel and ethinylestradiol interaction/control ratios confirmed the absence of pharmacokinetic interactions. CONCLUSION: The results from these studies demonstrate the absence of pharmacokinetic interactions between etonogestrel and ethinylestradiol released from NuvaRing and the oral antibiotics amoxicillin and doxycycline, suggesting that contraceptive efficacy would also be unaffected. PMID- 15828857 TI - Practical guidelines to interpret plasma concentrations of antiretroviral drugs. PMID- 15828858 TI - Three Prochlorococcus cyanophage genomes: signature features and ecological interpretations. AB - The oceanic cyanobacteria Prochlorococcus are globally important, ecologically diverse primary producers. It is thought that their viruses (phages) mediate population sizes and affect the evolutionary trajectories of their hosts. Here we present an analysis of genomes from three Prochlorococcus phages: a podovirus and two myoviruses. The morphology, overall genome features, and gene content of these phages suggest that they are quite similar to T7-like (P-SSP7) and T4-like (P-SSM2 and P-SSM4) phages. Using the existing phage taxonomic framework as a guideline, we examined genome sequences to establish "core" genes for each phage group. We found the podovirus contained 15 of 26 core T7-like genes and the two myoviruses contained 43 and 42 of 75 core T4-like genes. In addition to these core genes, each genome contains a significant number of "cyanobacterial" genes, i.e., genes with significant best BLAST hits to genes found in cyanobacteria. Some of these, we speculate, represent "signature" cyanophage genes. For example, all three phage genomes contain photosynthetic genes (psbA, hliP) that are thought to help maintain host photosynthetic activity during infection, as well as an aldolase family gene (talC) that could facilitate alternative routes of carbon metabolism during infection. The podovirus genome also contains an integrase gene (int) and other features that suggest it is capable of integrating into its host. If indeed it is, this would be unprecedented among cultured T7 like phages or marine cyanophages and would have significant evolutionary and ecological implications for phage and host. Further, both myoviruses contain phosphate-inducible genes (phoH and pstS) that are likely to be important for phage and host responses to phosphate stress, a commonly limiting nutrient in marine systems. Thus, these marine cyanophages appear to be variations of two well-known phages-T7 and T4-but contain genes that, if functional, reflect adaptations for infection of photosynthetic hosts in low-nutrient oceanic environments. PMID- 15828859 TI - A combinatorial code for splicing silencing: UAGG and GGGG motifs. AB - Alternative pre-mRNA splicing is widely used to regulate gene expression by tuning the levels of tissue-specific mRNA isoforms. Few regulatory mechanisms are understood at the level of combinatorial control despite numerous sequences, distinct from splice sites, that have been shown to play roles in splicing enhancement or silencing. Here we use molecular approaches to identify a ternary combination of exonic UAGG and 5'-splice-site-proximal GGGG motifs that functions cooperatively to silence the brain-region-specific CI cassette exon (exon 19) of the glutamate NMDA R1 receptor (GRIN1) transcript. Disruption of three components of the motif pattern converted the CI cassette into a constitutive exon, while predominant skipping was conferred when the same components were introduced, de novo, into a heterologous constitutive exon. Predominant exon silencing was directed by the motif pattern in the presence of six competing exonic splicing enhancers, and this effect was retained after systematically repositioning the two exonic UAGGs within the CI cassette. In this system, hnRNP A1 was shown to mediate silencing while hnRNP H antagonized silencing. Genome-wide computational analysis combined with RT-PCR testing showed that a class of skipped human and mouse exons can be identified by searches that preserve the sequence and spatial configuration of the UAGG and GGGG motifs. This analysis suggests that the multi component silencing code may play an important role in the tissue-specific regulation of the CI cassette exon, and that it may serve more generally as a molecular language to allow for intricate adjustments and the coordination of splicing patterns from different genes. PMID- 15828860 TI - A new yeast poly(A) polymerase complex involved in RNA quality control. AB - Eukaryotic cells contain several unconventional poly(A) polymerases in addition to the canonical enzymes responsible for the synthesis of poly(A) tails of nuclear messenger RNA precursors. The yeast protein Trf4p has been implicated in a quality control pathway that leads to the polyadenylation and subsequent exosome-mediated degradation of hypomethylated initiator tRNAMet (tRNAiMet). Here we show that Trf4p is the catalytic subunit of a new poly(A) polymerase complex that contains Air1p or Air2p as potential RNA-binding subunits, as well as the putative RNA helicase Mtr4p. Comparison of native tRNAiMet with its in vitro transcribed unmodified counterpart revealed that the unmodified RNA was preferentially polyadenylated by affinity-purified Trf4 complex from yeast, as well as by complexes reconstituted from recombinant components. These results and additional experiments with other tRNA substrates suggested that the Trf4 complex can discriminate between native tRNAs and molecules that are incorrectly folded. Moreover, the polyadenylation activity of the Trf4 complex stimulated the degradation of unmodified tRNAiMet by nuclear exosome fractions in vitro. Degradation was most efficient when coupled to the polyadenylation activity of the Trf4 complex, indicating that the poly(A) tails serve as signals for the recruitment of the exosome. This polyadenylation-mediated RNA surveillance resembles the role of polyadenylation in bacterial RNA turnover. PMID- 15828862 TI - Re: A case of a slipped medial rectus muscle after strabismus surgery. PMID- 15828861 TI - Re: Inferior oblique overaction/contracture following retrobulbar anesthesia for cataract extraction with a positive Bielschowsky Head Tilt Test to the contralateral shoulder. A report of one case. PMID- 15828863 TI - On the sagging of medical professionalism. PMID- 15828864 TI - New ethical minefield: drugs to boost memory and sharpen attention. PMID- 15828865 TI - The optometrization of ophthalmology. The association for research in vision and optometry? Get serious! Myopia is environmental but non-treatment is the standard and this is news? PMID- 15828866 TI - Two horizontal rectus eye muscle surgery combined with botulinum toxin for the treatment of very large angle esotropia. A pilot study. AB - PURPOSE: To evaluate the effectiveness of a proposed new protocol for the primary treatment for very large angle esotropia: two muscle horizontal rectus muscle surgery with simultaneous botulinum toxin A injection in a small pilot study. METHODS: Eight patients who had esotropia at near (ET') greater than 60 prism diopters (in actuality 70 to 100 prism diopters ET') underwent 2 muscle horizontal rectus surgery with simultaneous botulinum toxin A injection of the medial rectus intraoperatively. This was the only surgical procedure for all patients included in this report. Seven patients underwent bilateral medial rectus recession and bilateral injection, and one patient underwent a unilateral medial rectus recession / lateral rectus resection procedure with unilateral medial rectus injection. RESULTS: Postoperatively, 6 of the 8 patients demonstrated residual esotropia at near of less than 10 prism diopters and were considered "successful" by the conventional criteria of binocular alignment within 8 prism diopters of orthotropia. Two undercorrections occurred in patients with 100 and 85 prism diopters of preop ET' respectively. But 3 other patients with such large deviations had satisfactory results. All patients and families were satisfied with postoperative binocular alignment, so no further surgery was undertaken. The patient who underwent unilateral surgery had the least surgical effect and was the largest undercorrection, probably because only one medial rectus received a Botox injection. Considering only the bilateral cases, results were "successful" in 6 of 7 cases. Most patients suffered an extended period of Botox induced exotropia in the postop' period before recovery from the paresis. One patient had a transient, successfully treated, postoperative strabismic amblyopia while exotropic. CONCLUSIONS: Bilateral medial rectus recession with simultaneous botulinum injection is a safe and effective primary surgical procedure for very large angle esotropia. A more extensive study is indicated to confirm these findings. PMID- 15828867 TI - A study to determine: should conventional amounts of eye muscle surgery for horizontal binocular deviations be changed when oblique muscle weakening procedures are simultaneously performed? AB - PURPOSE: To investigate the final surgical outcome in horizontal strabismus correction, measured in primary position of gaze, in two situations: 1. When surgery was performed only on the horizontal rectus muscles; and 2. When inferior or superior oblique muscle weakening procedures were simultaneously performed. METHODS: Two hundred thirty cases were analyzed retrospectively: 172 esotropias (110 without oblique overaction; 23 with inferior oblique overaction; and 39 with superior oblique overaction), and 58 exotropias (25 without oblique overaction; 19 with inferior oblique overaction; and 14 with superior oblique overaction). Initial and final deviations, as well as the amount of correction achieved, was compared in six groups. RESULTS: No differences in outcomes and final results were found between any of the major or minor groups of patients. CONCLUSIONS: Oblique muscle weakening, in combination with horizontal muscle strabismus surgery did not affect the final results of the horizonal surgery in primary position. This study did suggest that there might not be a need for increasing or decreasing amounts of surgery on the horizontal rectus muscles to correct eso- or exotropia when oblique weakening procedures are included. PMID- 15828868 TI - A case of cyclic superior oblique paresis. AB - BACKGROUND: Cyclic strabismus, which generally has a 24 hours on-off cycle, is an unusual but interesting form of ocular motor disorder. Cyclic deviations are characterized by periods of significant deviation followed by periods of strict orthotropia. In the orthotropic periods, binocular sensoriality and motricity are normal. During the symptomatic phase, in which the deviation is usually significant and permanent, certain sensory anomalies can be found. Although most referred cases are cyclic esotropia, other cyclic oculomotor disorders have been described. In this report, we present a case with a 24 hours on-off cycle superior oblique paresis that resolved with surgery performed during the alternate day rhythm period. CASE REPORT: A 43 year old woman developed an alternate day hypertropia with bothersome diplopia. During the 24 hour "on" cycles she showed a unilateral left superior oblique paresis. The deviation had onset spontaneously 10 years ago with no previous history of strabismus, surgery or trauma. Following left inferior oblique recession and right inferior rectus recession surgery, the eyes have been straight and the cyclic pattern abolished. CONCLUSION: Cyclic vertical deviation due to a superior oblique paresis can be operated on by means of current surgical procedures in the course of the on-off cycle stage; i.e., before the cyclic pattern has been abolished and the deviation becoming permanent. PMID- 15828869 TI - Retinal nerve fiber analysis in subjects with hyperopia and anisometropic amblyopia. AB - PURPOSE: To determine the changes in retinal nerve fiber layer thickness in subjects with high hyperopia and anisometropic amblyopia. METHODS: Retinal nerve fiber layer thickness was measured with scanning laser polarimetry and confocal scanning laser tomography in subjects with hyperopia (n=18) with a mean age of 16.6 +/-3.1 years and anisometropic amblyopia (n=14) with a mean age of 17.4 +/ 3.8 years. A control group consisted of 25 healthy emmetropic subjects with a mean age of 25.7 +/-11.8 years. In all subjects 3 measurements with 2 different techniques were obtained and the average values for each parameter were analyzed and compared between the groups. RESULTS: Heidelberg Retinal Tomography revealed a smaller disc area and cup/disc ratio (C/D) in the hyperopic and amblyopic subjects in comparison to the control group, however there was no statistically significant difference between the hyperopia and amblyopia groups. Increased nerve fiber layer thickness in these groups had no statistical significance. There was also no statistically significant difference in various parameters in the measurements taken with scanning laser polarimetry. CONCLUSION: Refractive errors may change the quantitative assessment of optic nerve head topography and retinal nerve fiber layer thickness. There is no associated nerve fiber layer changes in anisometropic amblyopic subjects in comparison to the control group and the subjects with high hyperopia. PMID- 15828870 TI - Characterization of the Bacillus subtilis GTPase YloQ and its role in ribosome function. AB - We present an analysis of the cellular phenotype and biochemical activity of a conserved bacterial GTPase of unknown function (YloQ and YjeQ in Bacillus subtilis and Escherichia coli respectively) using a collection of antibiotics of diverse mechanisms and chemical classes. We created a yloQ deletion strain, which exhibited a slow growth phenotype and formed chains of filamentous cells. Additionally, we constructed a conditional mutant in yloQ, where growth was dependent on inducible expression from a complementing copy of the gene. In phenotypic studies, depletion of yloQ sensitized cells to antibiotics that bind at the peptide channel or peptidyl transferase centre, providing the first chemical genetic evidence linking this GTPase to ribosome function. Additional experiments using these small-molecule probes in vitro revealed that aminoglycoside antibiotics severely affected a previously characterized ribosome associated GTPase activity of purified, recombinant YjeQ from E. coli. None of the antibiotics tested competed with YjeQ for binding to 30 or 70 S ribosomes. A closer examination of YloQ depletion revealed that the polyribosome profiles were altered and that decreased expression of YloQ led to the accumulation of ribosomal subunits at the expense of intact 70 S ribosomes. The present study provides the first evidence showing that YloQ/YjeQ may be involved in several areas of cellular metabolism, including cell division and ribosome function. PMID- 15828871 TI - Growth factors induce differential phosphorylation profiles of the Hrs-STAM complex: a common node in signalling networks with signal-specific properties. AB - Hrs (hepatocyte growth factor-regulated tyrosine kinase substrate) and STAM (signal-transducing adaptor molecule) form a heterodimeric complex that associates with endosomal membranes and is tyrosine-phosphorylated in response to a variety of growth factors including EGF (epidermal growth factor), HGF (hepatocyte growth factor) and PDGF (platelet-derived growth factor). Phosphorylation of the Hrs-STAM complex requires receptor endocytosis. We show that an intact UIM (ubiquitin interaction motif) within Hrs is a conserved requirement for Hrs phosphorylation downstream of both EGF and HGF stimulations. Consistent with this, expression of a dominant-negative form of the E3 ubiquitin ligase, c-Cbl, inhibits EGF- and HGF-dependent Hrs phosphorylation. Despite this conservation, kinase inhibitor profiles using PP1 (4-amino-5-(4-methylphenyl)-7 (t-butyl)pyrazolo[3,4-d]pyrimidine) and SU6656 indicate that distinct non receptor tyrosine kinases couple EGF, HGF and PDGF stimulation with the tyrosine phosphorylation of the Hrs-STAM complex. Crucially, analysis with phospho specific antibodies indicates that these kinases generate a signal-specific, combinatorial phosphorylation profile of the Hrs-STAM complex, with the potential of diversifying tyrosine kinase receptor signalling through a common element. PMID- 15828872 TI - Characterization of a novel Foxa (hepatocyte nuclear factor-3) site in the glucagon promoter that is conserved between rodents and humans. AB - The pancreatic islet hormone glucagon stimulates hepatic glucose production and thus maintains blood glucose levels in the fasting state. Transcription factors of the Foxa [Fox (forkhead box) subclass A; also known as HNF-3 (hepatocyte nuclear factor-3)] family are required for cell-specific activation of the glucagon gene in pancreatic islet alpha-cells. However, their action on the glucagon gene is poorly understood. In the present study, comparative sequence analysis and molecular characterization using protein-DNA binding and transient transfection assays revealed that the well-characterized Foxa-binding site in the G2 enhancer element of the rat glucagon gene is not conserved in humans and that the human G2 sequence lacks basal enhancer activity. A novel Foxa site was identified that is conserved in rats, mice and humans. It mediates activation of the glucagon gene by Foxa proteins and confers cell-specific promoter activity in glucagon-producing pancreatic islet alpha-cell lines. In contrast with previously identified Foxa-binding sites in the glucagon promoter, which bind nuclear Foxa2, the novel Foxa site was found to bind preferentially Foxa1 in nuclear extracts of a glucagon-producing pancreatic islet alpha-cell line, offering a mechanism that explains the decrease in glucagon gene expression in Foxa1-deficient mice. This site is located just upstream of the TATA box (between -30 and -50), suggesting a role for Foxa proteins in addition to direct transcriptional activation, such as a role in opening the chromatin at the start site of transcription of the glucagon gene. PMID- 15828873 TI - Mobilization of the iron centre in IscA for the iron-sulphur cluster assembly in IscU. AB - The biogenesis of iron-sulphur clusters requires the co-ordinated delivery of both iron and sulphur. It is now clear that sulphur in iron-sulphur clusters is derived from L-cysteine by cysteine desulphurases. However, the iron donor for the iron-sulphur cluster assembly still remains elusive. Our previous studies indicated that Escherichia coli IscA, a member of the iron-sulphur cluster assembly machinery, is an iron-binding protein that can provide iron for the iron sulphur cluster assembly in a proposed scaffold IscU. To determine how the iron centre in IscA is transferred for the iron-sulphur cluster assembly in IscU, we explore the mobility of the iron centre in IscA. The UV-visible and EPR measurements show that L-cysteine, but not IscU, is able to mobilize the iron centre in IscA and make the iron available for the iron-sulphur cluster assembly in IscU. Other related biological thiols such as N-acetyl-L-cysteine or reduced glutathione have no effect on the iron centre of IscA, suggesting that L-cysteine is unique in mobilizing the iron centre of IscA. Nevertheless, L-cysteine alone is not sufficient to transfer the iron from IscA to IscU. Both L-cysteine and cysteine desulphurase (IscS) are required for the IscA-mediated assembly of iron sulphur clusters in IscU. The results suggest that L-cysteine may have two distinct functions in the biogenesis of iron-sulphur clusters: to mobilize the iron centre in IscA and to provide sulphur via cysteine desulphurase (IscS) for the iron-sulphur cluster assembly in IscU. PMID- 15828874 TI - Site-specific base changes in the coding or promoter region of the human beta- and gamma-globin genes by single-stranded oligonucleotides. AB - SSOs (single-stranded oligonucleotides) can mediate site-specific alteration of base-pairs in episomal and chromosomal target genes in mammalian cells. The TNE (targeted nucleotide exchange) can result in either repair or mutation of a gene sequence and is mediated through endogenous DNA repair pathway(s). Thus the approach provides a technique for the treatment of monogenic disorders associated with specific point mutations such as SCD (sickle cell disease). We studied the potential application of SSOs for SCD by introducing either an A to T substitution at the sixth codon of the human beta-globin gene (sickle locus) or a C to G mutation at -202 of the Ggamma-globin gene promoter region. The latter TNE is an alternative strategy to ameliorate the clinical manifestations of sickle cell anaemia by re-activating fetal haemoglobin gene expression in adult erythrocytes. A sensitive and valid PCR assay system was developed, which allows detection of point mutations as low as 0.01% at these sites. Using this system, TNE between 0.01 and 0.1% at the sickle locus or gamma-globin gene promoter region was detected after transfection with SSOs in cultured human cell lines. TNE in the Ggamma-globin promoter region exhibited varying degrees of strand bias that was dependent on SSO design and the cell's DNA mismatch repair activity. The results suggest that the endogenous DNA repair machinery may permit SSO correction of the sickle defect by modification of the beta- and/or gamma-globin genes. PMID- 15828875 TI - The clinical implications of cumulative right ventricular pacing in the multicenter automatic defibrillator trial II. AB - INTRODUCTION: This study was designed to assess whether right ventricular pacing in the implantable cardioverter defibrillator (ICD) arm of the Multicenter Automatic Defibrillator Implantation Trial (MADIT) II was associated with an unfavorable outcome. METHODS AND RESULTS: Data on the number of ventricular paced beats were available in 567 (76%) of 742 MADIT II patients with ICDs. The number of ventricular paced beats over the total number of beats showed a bimodal distribution with patients being predominantly paced or nonpaced. Therefore, patients were dichotomized at 0-50% and 51-100% of cumulative pacing with median pacing rate 0.2% and 95.6%, respectively. Endpoints included new or worsening heart failure, appropriate ICD therapy for VT/VF, and the combined endpoint of heart failure or death. Clinical features associated with frequent ventricular pacing included age >or=65 years, advanced NYHA heart failure class, LVEF < 0.25, first degree AV and bundle branch block, and amiodarone use. During follow-up, 119 patients (21%) had new or worsened heart failure, 130 (23%) had new or worsened heart failure or death, and 142 (25%) had appropriate therapy for VT/VF. In comparison to patients with infrequent pacing, those with frequent pacing had significantly higher risk of new or worsened heart failure (hazard ratio = 1.93; P = 0.002) and VT/VF requiring ICD therapy (HR = 1.50; P = 0.02). CONCLUSIONS: Patients in MADIT II who were predominantly paced had a higher rate of new or worsened heart failure and were more likely to receive therapy for VT/VF. These results suggest the deleterious consequences of RV pacing, particularly in the setting of severe LV dysfunction. PMID- 15828876 TI - Cryocatheter ablation of right ventricular outflow tract tachycardia. AB - INTRODUCTION: Cryocatheter techniques have been successfully applied to treat supraventricular tachycardia but there are no reports on their value in treating ventricular tachycardia (VT). We present our initial experience with cryocatheter ablation of right ventricular outflow tract (RVOT) tachycardia. METHODS AND RESULTS: Cryocatheter ablation was attempted in 14 patients (13 females, age 45.9 +/- 12.7 years) who were highly symptomatic due to frequent monomorphic ventricular extrasystole (VES) or nonsustained VT originating within the RVOT. A 9-Fr, 8-mm-tip cryocatheter was used for both mapping and ablation. Cryoablation was started after localizing the arrhythmic focus by pace and activation mapping. Ablation success, defined by complete disappearance of target VES/VT acutely and during a follow-up of 9.3 +/- 1.4 weeks, was achieved in 13 of 14 patients. Ablation was successful with local activation times of 35 +/- 4 ms, 5.8 +/- 3.3 applications, 18.8 +/- 7.5 minutes total cryo time, 9.4 +/- 4.2 minutes fluoroscopy time, and 66.9 +/- 26.1 minutes total procedure time, the latter two measures showing a reduction with number of patients treated. Three patients reported slight pain related to local pressure of the catheter on the RVOT wall. No pain was described related to delivery of cryothermal energy. CONCLUSIONS: Initial experience shows that focal VES/VT originating in the RVOT can be successfully treated using cryocatheter ablation. Acute and short term success rates, fluoroscopy times, and duration of procedure are comparable to conventional ablation techniques. A major advantage seems to be the virtual absence of ablation related pain. PMID- 15828877 TI - Return to the ice age? PMID- 15828878 TI - Effects of psychologic stress on repolarization and relationship to autonomic and hemodynamic factors. AB - INTRODUCTION: Psychological stress can precipitate ventricular arrhythmias in patients with ICDs, as well as sudden death. However, the physiologic pathways remain unknown. We sought to determine whether psychological stress induced in the laboratory setting alters indices of repolarization associated with arrhythmogenesis. METHODS AND RESULTS: Patients with ICDs and a history of ventricular arrhythmia underwent ambulatory ECG monitoring during a laboratory mental stress protocol (anger recall and mental arithmetic). Continuous changes in repolarization indices which have correlated with temporal and spatial myocardial heterogeneity of repolarization, including T-wave alternans (TWA), T wave amplitude (Tamp), and T-wave area (Tarea) were analyzed in the time domain. In the 33 patients (85% male, 88% with coronary artery disease, mean ejection fraction 30%), norepinephrine, epinephrine, BP, and HR increased during mental stress. TWA increased from 22 (interquartile range 16-27) at baseline to 29 (21 38) uV during mental stress (P < 0.001). Changes in TWA correlated with changes in HR, systolic BP, and catecholamines. Tamp and Tarea also increased with mental stress (P < 0.01) but did not correlate with changes in other variables. CONCLUSION: Psychological stress increased TWA, Tamp, and Tarea. Autonomically mediated repolarization changes may be a pathophysiologic link between emotion and arrhythmia in susceptible patients. PMID- 15828879 TI - Clinical and electrophysiological characteristics of Brugada syndrome caused by a missense mutation in the S5-pore site of SCN5A. AB - Brugada syndrome is an inherited cardiac disorder caused by mutations in the SCN5A gene encoding the cardiac sodium channel alpha-subunit, and potentially leads to ventricular fibrillation and sudden death. We report a case of a novel SCN5A mutation associated with Brugada syndrome. A 51-year-old man suffered from recurrent nocturnal syncopal attacks due to polymorphic ventricular tachycardia. His electrocardiogram showed ST-segment elevation in V1-V3 leads, but there was no evidence of structural heart disease. DNA sequence analysis of SCN5A in this patient revealed a missense mutation (R282H) in the S5-pore region of domain I. This mutational change was not present in 100 healthy Japanese controls. In the patient's family, a 36-year-old brother had died suddenly. Genetic analysis identified two other carriers of the R282H mutation, who had ST-segment elevation and slightly increased QRS widths, but they experienced no syncopal episodes or ventricular fibrillation. Electrophysiological investigation of the R282H mutant channel expressed in cultured cells showed a severe reduction in sodium current density and a mild positive shift of activation curve. R282H did not enhance intermediate inactivation. Single-channel conductance of R282H was slightly decreased compared with WT. The electrophysiological characteristics of the R282H channel are suggested to be closely related to the clinical phenotype of Brugada syndrome. PMID- 15828880 TI - Measuring patient acceptance of implantable cardiac device therapy: initial psychometric investigation of the Florida Patient Acceptance Survey. AB - INTRODUCTION: Patient acceptance of implantable device therapy has been established as an important outcome but the operationalization and validation of a measure of patient acceptance of implantable device therapy has not been fully completed. This study sought to validate a new measure of patient acceptance of cardiac implantable devices called the Florida Patient Acceptance Survey (FPAS). METHODS: The sample consisted of implantable cardioverter defibrillator (ICD; n = 58), and implantable atrioverter defibrillator (ICD-AT; n = 96), and pacemaker (PM, n = 84) patients. Mean age of all participants was 69 years; they were mostly male (62%) and married (75%). The final FPAS comprised 15 items with four consistent factors: Return to Function, Device-Related Distress, Positive Appraisal, and Body Image Concerns. RESULTS: The total FPAS demonstrated good internal consistency (Cronbach's alpha = 0.83), and internal consistency for each of the subscales ranged from 0.74 to 0.89. The FPAS demonstrated convergent, divergent, and discriminant validity when compared to other self-report measures of QOL, atrial symptoms, depression, and anxiety. A total FPAS score can be formed and between group comparisons with this sample indicated that ICD patients report a high level of acceptance (mean = 76), ICD-AT patients report a significantly higher level of acceptance (mean = 81.1), and PM patients reported the highest level of patient acceptance (mean = 85.4) of these implantable device groups. CONCLUSION: This initial psychometric investigation of the FPAS suggests that the FPAS may be useful in both clinical and research settings to assess patient acceptance of implantable cardiac devices. PMID- 15828881 TI - Emphasizing quality in qualitative research: devices to control arrhythmias. PMID- 15828882 TI - Short QT syndrome and atrial fibrillation caused by mutation in KCNH2. AB - BACKGROUND: The short QT syndrome is a newly described clinical entity characterized by the presence of a short QT interval associated with cardiac tachyarrhythmias including sudden cardiac death at a young age in otherwise healthy individuals. A genetic basis has been identified linking the disease to mutations in KCNH2 in the familial forms and a mutation in KCNQ1 in a sporadic form of the disease. METHODS AND RESULTS: We identified a family with short QT syndrome with a high incidence of paroxysmal atrial fibrillation in their members and no known history of sudden cardiac death. QT interval ranged from 225 to 240 ms within normal heart rate ranges in the affected individuals. Programmed electrical stimulation (PES) was performed in all affected members, which revealed a remarkably short atrial and ventricular refractory period, and inducibility of atrial and ventricular fibrillation. Treatment with propafenone has maintained the individuals free of atrial fibrillation to date. Genetic analysis identified a missense mutation (C to G substitution at nucleotide 1764) which resulted in the amino acid change (N588K) in KCNH2. This mutation had been previously described in two other families with a high incidence of sudden cardiac death. CONCLUSIONS: Our study confirms that N588K is a hotspot for familial form of the short QT syndrome. The disease is clinically heterogeneous, as indicated by the fact that, in the three families with the same mutation, there is a wide range of symptoms, varying from atrial to ventricular fibrillation and sudden death. While the implantation of a defibrillator appears warranted due to the inducibility at PES, the clinical follow-up provides indication that the class Ic agent propafenone could be effective to prevent episodes of paroxysmal atrial fibrillation. PMID- 15828883 TI - Short QT interval and short QT syndromes. PMID- 15828884 TI - Induction ovens and electromagnetic interference: what is the risk for patients with implantable cardioverter defibrillators? AB - Electromagnetic fields may interfere with normal implantable cardioverter defibrillator (ICD) function. Although the devices are effectively shielded and use exclusively bipolar leads, electromagnetic interference (EMI) remains a concern when patients are exposed to several household appliances. The aim of this study was to evaluate potential EMI risk of induction ovens, which are increasingly common in private households. In vitro measurements of an induction oven for private households GK 43 TI (V-Zug, Inc., Zug, Switzerland) showed that heating is regulated by increasing operating time from level 1 (100 ms/sec) to 5 (continuous operation). From levels 5 to 9 the magnetic field increases. Nineteen patients with left-sided implants of single- and dual-chamber ICD systems (8 Medtronic, 7 Guidant, and 4 St. Jude Medical) (18 males, 1 female), age (mean +/- SEM) 58 +/- 3 years, were included in this study. All patients were examined in standing position, bent over the cooking pot (minimal distance to the induction coil 25 cm), and with the cooking pot put eccentrically over the induction field at three different cooking levels (level 2, 5, and 9). The tests were repeated touching the cooking pot with one hand. Ventricular sensitivity was left unchanged. Ventricular tachycardia therapies were turned off in Medtronic and Guidant devices and ventricular sensing was continuously monitored in St. Jude Medical devices during testing. Interrogation of the devices after exposure did not show any inappropriate tachycardia detection, oversensing, or reprogramming. In conclusion, ICD patients can be reassured that EMI is unlikely to affect their devices if induction ovens are used in their kitchens. PMID- 15828885 TI - The inferior right atrial isthmus: further architectural insights for current and coming ablation technologies. AB - BACKGROUND: Although linear ablation of the right atrial isthmus in patients with isthmus-dependent atrial flutter can be highly successful, recurrences and complications occur in some patients. Our study provides further morphological details for a better understanding of the structure of the isthmus. METHODS AND RESULTS: We examined the isthmic area in 30 heart specimens by dissection, histology, and scanning electron microscopy. This area was bordered anteriorly by the hinge of the tricuspid valve and posteriorly by the orifice of the inferior caval vein. With the heart in attitudinal orientation, we identified and measured the lengths of three levels of isthmus: paraseptal (24 +/- 4 mm), central (19 +/- 4 mm), and inferolateral (30 +/- 3 mm). Comparing the three levels, the central isthmus had the thinnest muscular wall and the paraseptal isthmus the thickest wall. At all three levels, the anterior part was consistently muscular whereas the posterior part was composed of mainly fibro-fatty tissue in 63% of hearts. The right coronary artery was less than 4 mm from the endocardial surface of the inferolateral isthmus in 47% of hearts. Inferior extensions of the atrioventricular node were present in the paraseptal isthmus in 10% of hearts, at 1-3 mm from the endocardial surface. CONCLUSIONS: The thinner wall and shorter length of the central isthmus together with its distance from the right coronary artery, and nonassociation with the atrioventricular node or its arterial supply, should make it the preferred site for linear radiofrequency ablation. PMID- 15828886 TI - More on isthmus anatomy for safety and efficacy. PMID- 15828887 TI - Morphology of the thoracic veins and left atrium in paroxysmal atrial fibrillation initiated by superior caval vein ectopy. AB - INTRODUCTION: The structural changes of the superior caval vein, pulmonary veins, and left atrium in atrial fibrillation initiated by superior caval vein ectopy have not been reported. METHODS AND RESULTS: Nine patients with atrial fibrillation initiated by superior caval vein ectopic beats (male = 5, 54 +/- 10 years) and 15 control (n = 15, male = 10, 52 +/- 8 years) without any cardiac arrhythmias were included in this study. Using gadolinium-enhanced magnetic resonant angiography with three-dimensional reconstruction, the parameters of the superior caval vein morphology (length, various diameters, area, eccentricity, and volume) were measured. The morphological parameters of the four pulmonary veins (diameter, ostial area, and eccentricity) were also measured at the pulmonary vein-left atrial junction in an oblique sagittal section from the multiple-plane reconstruction images. The left atrial diameters and volume were measured. The different morphological parameters were compared between the two groups. The patients with atrial fibrillation initiated by superior caval vein ectopic beats exhibited a more eccentric structure of the second part of the superior caval vein as compared to the control group. All the ectopic beats initiating atrial fibrillation were located in the second part of the superior caval vein. Furthermore, the patients with atrial fibrillation initiated by superior caval vein ectopic beats had a larger superior caval vein volume, left atrial volume, and pulmonary vein size, and more eccentric pulmonary vein ostia than the controls. CONCLUSION: Structural changes of the superior caval vein were demonstrated in the patients with atrial fibrillation initiated by superior caval vein ectopic beats. These findings might explain the arrhythmogenic mechanism of atrial fibrillation initiated by superior caval vein ectopy. PMID- 15828888 TI - Delayed after depolarization-mediated triggered activity associated with slow calcium sequestration near the endocardium. AB - INTRODUCTION: Previously, we have shown that cells near the endocardium are more prone to elevated diastolic intracellular calcium levels than cells near the epicardium. The arrhythmogenic consequence of such regional differences in calcium handling is not clear. METHODS AND RESULTS: Using optical mapping techniques, calcium transients and action potentials were recorded simultaneously from ventricular sites across the transmural wall of the arterially perfused canine left ventricular wedge preparation during control conditions, and under conditions of increased calcium entry (I(K) blockade and beta-adrenergic stimulation). Under conditions of enhanced calcium entry, the decay of the calcium transient and diastolic calcium levels during rapid pacing were slower (38%, P < 0.01) and higher (215%, P < 0.02), respectively, near (within approximately 3 mm) the endocardium compared to the epicardium (n = 9). Immediately after termination of rapid pacing under conditions of increased calcium entry, ectopic activity and simultaneous delayed after depolarizations and spontaneous calcium release events were observed. Over all experiments, ectopic activity occurred more frequently closer to the endocardium compared to the epicardium. CONCLUSIONS: Under conditions of enhanced calcium entry, myocytes closer to the endocardium exhibit a higher level of diastolic calcium and greater ectopic activity compared to the epicardium. We show for the first time simultaneous delayed after depolarization and spontaneous calcium release events from myocytes in a normally coupled multicellular preparation. These data combined suggest that myocytes near the endocardium are more susceptible to calcium-mediated triggered activity. PMID- 15828889 TI - Regional variation in capture of fibrillating swine left ventricle during electrical stimulation. AB - INTRODUCTION: While it has been shown that electrical stimulation can capture a region of myocardium during ventricular fibrillation (VF), the ideal location to stimulate to maximize capture of the fibrillating in vivo left ventricle (LV) is not known. We previously demonstrated a mean directionality to the propagation of VF wavefronts in swine from posterior to anterior LV. We hypothesized that this directionality of VF wavefronts would affect capture of the LV epicardium while stimulating during VF. METHODS AND RESULTS: In seven open-chest swine, during different VF episodes, electrical stimulation was performed singly or simultaneously from two lines of 26 epicardial electrodes, one on the posterior LV adjacent to the posterior descending coronary artery and another on the anterior LV adjacent to the left anterior descending coronary artery. Mapping was performed between the line of stimulating electrodes with 768 recording electrodes 2-mm apart. The incidence and extent of epicardium captured by stimulation through the lines of stimulating electrodes were determined in the mapped region. Capture occurred during 67% of 78 VF episodes. Capture from the posterior LV line was achieved in 88% of the episodes and from the anterior LV line in 44% of the episodes (P = 0.001). The maximum amount of myocardium captured was also much greater for stimulating from the posterior as compared to the anterior LV line (232 +/- 168 mm(2) vs 64 +/- 124 mm(2), P = 0.003). A significant part of the variability in capture was related to the direction of the mean VF wavefront velocity vector in each animal (r = 0.84, P < 0.05). CONCLUSION: Electrical stimulation from the posterior LV resulted in a greater incidence and extent of LV capture than stimulation from the anterior LV. A significant component of the variability in capture is related to the mean direction of VF wavefronts. PMID- 15828890 TI - Role of streptomycin-sensitive stretch-activated channel in chest wall impact induced sudden death (commotio cordis). AB - INTRODUCTION: Deaths secondary to low-energy impacts to the precordium in young individuals (commotio cordis) have been reported with increasing frequency. In a swine model, baseball impacts induce ventricular fibrillation when directed at the center of the left ventricle during the vulnerable portion of repolarization just prior to the T-wave peak. It has been hypothesized that activation of stretch-sensitive channels could be crucial for this electrophysiological phenomenon. In this study, a nonselective stretch-activated cation channel was pharmacologically blocked prior to chest blows to determine whether this channel represents a possible pathway by which commotio cordis events occur. METHODS: In a randomized and blinded experiment, 12 swine (mean 17.1 +/- 2.5 kg) received either 2-g streptomycin intramuscularly (mean serum concentration 115 +/- 18 muM) or sterile water prior to chest impact. Each animal received six precordial impacts with a baseball propelled at 40 mph. RESULTS: There was no significant difference in the frequency of induced VF in the animals administered streptomycin (10 of 19 impacts: 53%) compared to those control animals receiving only sterile water (10 of 31: 32%) (P = 0.15). However, the magnitude of ST segment elevation was less in the streptomycin-treated animals (19 +/- 19 mV) versus controls (61 +/- 46 mV) (P = 0.015). CONCLUSION: Streptomycin did not alter the frequency of ventricular fibrillation in our commotio cordis model, indicating that the stretch-activated channel is not implicated in the genesis of chest blow-induced cardiac arrest. However, streptomycin did reduce ST elevation following impact suggesting that the stretch-activated channel may play a role in ST segment elevation following chest wall blows. PMID- 15828891 TI - Cardiac resynchronization therapy: retiming the failing right ventricle. AB - Cardiac resynchronization therapy (CRT) improves symptoms, reduces hospitalization, and may decrease mortality in patients with moderate/severe heart failure and left bundle branch block. Whether CRT may have a role in the management of patients with adult congenital heart disease and a failing right (systemic) ventricle is unknown. We report the case of an adult patient with transposition of the great arteries and previous Mustard's repair, who successfully underwent CRT using a hybrid transvenous/epicardial approach. Exercise tolerance improved, right ventricular (systemic) ejection fraction improved, diuretic requirements reduced, and renal function improved. CRT may offer a new therapeutic option for this patient population. PMID- 15828892 TI - Automatic impedance monitoring and patient alert feature in implantable cardioverter defibrillators: being alert for the unexpected! AB - Recent advances in implantable cardioverter defibrillator (ICD) technology have enabled implementation of an automatic monitoring feature in ICDs that provides daily measurements of several technical parameters such as battery status, pacing, and high-voltage impedance. The system alerts the patient with an audible alarm to contact the physician in case the measured parameters are not within normal limits. Early detection of intermittent and potentially serious complications justifies routine use of this feature. This report describes a patient with an ICD who died suddenly due to ventricular fibrillation, which was not appropriately treated by the device. The cause of device malfunction was most likely an intermittent lead fracture that was not detected by the automatic impedance monitoring and alert feature of this device. Based on these data, potential benefits and theoretical pitfalls of automatic impedance monitoring and alert features are discussed. PMID- 15828893 TI - Epicardial mapping and ablation techniques to control ventricular tachycardia. PMID- 15828894 TI - Combined P/S-HV ICD lead implantation via the coronary sinus in a patient with a prosthetic tricuspid valve. PMID- 15828895 TI - Transmural conduction is not a two-way street. PMID- 15828896 TI - How to apply and adapt international guidelines on bone metabolism and disease to the management of Japanese dialysis patients. PMID- 15828897 TI - Abstracts from the 3rd Slovenian Congress of Nephrology. PMID- 15828898 TI - Challenges in the therapy of secondary hyperparathyroidism. AB - Derangements in mineral metabolism are known to occur early in the course of chronic kidney disease (CKD). Recent clinical practice guidelines are designed to focus on the problem early in the course of kidney disease, when it is recommended to evaluate the levels of parathyroid hormone (PTH) and to try to intervene early if the levels are elevated. To begin early intervention for hyperparathyroidism in chronic kidney disease will require involvement of primary care physicians and other subspecialty groups to identify the patients at risk and begin to intervene with measures to control hyperparathyroidism and its consequences on mineral metabolism. It has recently been demonstrated that chronic kidney disease is a significant risk factor for vitamin D deficiency and since abnormalities in vitamin D metabolism are important in the generation of hyperparathyroidism, this is an issue that needs direct attention. Studies are needed to assess the effects of correcting this vitamin D deficiency in early CKD. As kidney disease progresses, efforts to control hyperparathyroidism will likely need to be intensified and several therapeutic options are available, such as phosphate binders, repletion of vitamin D, the use of active vitamin D sterols, or the use of vitamin D analogs. In addition, it is important to define the appropriate PTH values that need to be achieved to minimize complications on bone. Such studies are in progress at the present time to validate the current more specific PTH assays. Strict guidelines have been proposed for the management of bone and mineral metabolism in patients with CKD stage V on dialysis, and although these challenging recommendations were initially opinion-based, there is mounting evidence which provides confirmation of these targets as relevant. Treatment options for patients on dialysis involve the full spectrum of agents which include phosphate binders, active vitamin D sterols (often given parenterally), the use of calcimimetic agents, surgical parathyroidectomy, and evaluation of appropriate levels of dialysate calcium. Similar to early stages of CKD, studies are in progress to refine the PTH targets with the newer PTH assays. With increased focus on the complications of bone and mineral metabolism as part of the continuum of chronic kidney disease, and with a variety of new therapies available, it is anticipated that improved patient outcomes should be achievable in this patient group. PMID- 15828899 TI - Problems in adapting the K/DOQI guidelines for Japanese patients with end stage renal failure. AB - The National Kidney Foundation published The K/DOQI Clinical Practice Guidelines for Bone Metabolism and Disease in Chronic Kidney Disease. We should also consider differences in clinical features of management for renal osteodystrophy between Japan and the USA. The first problem is related to a difference in the method of Ca correction. The second problem is related to a difference in the timing of measurement. In the USA the timing of measurement varies with each institution. Briefly, it is performed at midweek (2 days after the last dialysis) in some of institutions, while at the beginning of next week (3 days after the last dialysis) in some others. In addition, differences in the dose and type of the calcium-containing phosphorus adsorbent and active vitamin D used should also be taken into consideration. PMID- 15828900 TI - Combination therapy with sevelamer hydrochloride and calcium carbonate in Japanese patients with long-term hemodialysis: alternative approach for optimal mineral management. AB - Calcium (Ca) overload by Ca-containing phosphorus (P) binder has been suggested to be implicated in the pathogenesis of soft tissue and vascular calcification, which contribute to increased morbidity and mortality of cardiovascular disease in patients undergoing dialysis. Recently, a noncalcium P binder, sevelamer hydrochloride (sevelamer), has become available in Japan. However, Japanese patients undergoing dialysis might be less tolerant of sevelamer treatment, and it is likely to cause hypocalcemia because their dietary Ca intake is less than that in European and American patients. We evaluated the effects of combination therapy with sevelamer and calcium carbonate (CC) on mineral metabolism in Japanese hemodialysis patients, as an alternative form of P management. A total of 210 hemodialysis patients were enrolled, and were given a small dose of sevelamer (0.75-1.5 g/day) on CC treatment. Sevelamer dose was gradually increased, while CC decreased during 24 weeks. Five patients discontinued sevelamer treatment because of severe constipation, anorexia, and parathyroidectomy for severe secondary hyperparathyroidism. After 24 weeks, the dose of sevelamer was significantly increased to 3.29 g/day (initial dose: 1.47 g/day), while CC was decreased by 54%. Adjusted serum Ca significantly decreased (9.63 +/- 0.57-9.45 +/- 0.67 mg/dL; P = 0.0012), although serum P increased (5.89 +/- 1.32-6.25 +/- 1.32 mg/dL; P = 0.017). Serum intact PTH (iPTH) significantly increased in patients with a low or normal iPTH level (< or =300 pg/mL), while it did not change in patients with secondary hyperparathyroidism (>300 pg/mL). The results suggest that the therapeutic regimen is more tolerant and reduces Ca load in Japanese hemodialysis patients while avoiding hypocalcemia. In addition, the mitigated Ca overload could improve PTH hyposecretion in patients with adynamic bone disease, which is associated with soft tissue calcification and higher mortality in uremia. PMID- 15828901 TI - The influence of dialysate calcium on the therapeutic effects of sevelamer hydrochloride in hemodialysis patients with secondary hyperparathyroidism under treatment of intravenous vitamin d metabolites. AB - The management of hyperphosphatemia is essential to treat secondary hyperparathyroidism and to prevent ectopic calcification. Sevelamer hydrochloride (sevelamer), a new phosphate binder that contains neither aluminum nor calcium, which could be theoretically beneficial for the management of hyperphosphatemia in dialysis patients with secondary hyperparathyroidism who are receiving intravenous vitamin D metabolites (maxacalcitol or calcitriol). To reduce calcium loads, a dialysate calcium concentration of 2.5 mEq/L is recommended by Kidney Disease Outcome Quality Initiative (K/DOQI) guidelines. In Japan, a dialysate calcium concentration of 3.0 mEq/L prevails. We investigated the influence of dialysate calcium on the therapeutic effect of sevelamer in 40 hemodialysis patients who are under treatment of intravenous vitamin D metabolites for secondary hyperparathyroidism (VD(+)) and compared the results with those of 41 patients who had not received vitamin D metabolites (VD(-)). Serum phosphorus and calcium-phosphorus products showed no significant change by sevelamer in either the VD(+) subgroup of patients receiving hemodialysis with dialysate calcium of 2.5 mEq/L (DCa2.5) or those receiving hemodialysis with dialysate calcium of 3.0 mEq/L (DCa3.0), while serum phosphorus and calcium-phosphorus products decreased in both the VD(-) subgroups. Serum calcium decreased in the DCa2.5 subgroup and did not change in the DCa3.0 subgroup in both the VD(+) and the VD(-) subjects. Parathyroid hormone and alkaline phosphatase increased in the DCa2.5 subgroup and did not change in the Ca 3.0 subgroup in the VD(+) subjects. Serum calcium decreased in both subgroups in the VD(-) subjects. Parathyroid hormone obtained after sevelamer administration in the VD(-) group was within the target range of the K/DOQI guidelines. In conclusion, the concomitant use of sevelamer as a phosphate binder and the dialysate of calcium concentration of 2.5 mEq/L have possibilities for worsening secondary hyperparathyroidism in patients receiving intravenous vitamin D. PMID- 15828902 TI - Is 2.5 mEq/L the optimal calcium concentration of dialysate in the use of sevelamer hydrochloride? A study of the dialysate calcium concentration recommended by K/DOQI guidelines. AB - We tested the effect of three different dialysate calcium concentrations on calcium-phosphorus metabolism during the use of sevelamer hydrochloride. After a calcium-containing phosphate binder was switched to sevelamer, the serum calcium, phosphorus, and intact parathyroid hormone levels and the markers of bone turnover were measured in the patients whose dialysate calcium concentrations were 2.5, 2.75, and 3.0 mEq/L. As a result, in the 2.75-mEq/L group, the serum calcium concentrations decreased and the intact parathyroid hormone level increased significantly. In the 2.5-mEq/L group, transient hypocalcemia occurred and the levels of both bone-alkaline phosphatase and osteocalcin increased. In the 3.0-mEq/L group, the serum calcium concentrations did not change significantly and only bone-alkaline phosphatase increased. If a calcium containing phosphate binder is completely switched to sevelamer, dialysis using a dialysate calcium concentration below 3.0 mEq/L may result in hypocalcemia and acceleration of bone turnover. PMID- 15828903 TI - Different routes bridging calcium in Japanese hemodialysis patients. AB - There is growing evidence that not only serum calcium concentration but also excess calcium load is associated with vascular calcification and mortality in hemodialysis patients. Calcium load in hemodialysis patients cumulatively comes from three different routes: oral intake of calcium including calcium-based phosphate binders, traffic of calcium from/to dialysate, and calcemic action of vitamin D. The K/DOQI guidelines recommend sevelamer hydrochloride instead of calcium-containing phosphate binders to control serum phosphate concentration. However, in Japan, both kinds of phosphate binders are used concomitantly, mainly because Japanese patients are prone to a higher incidence of sevelamer-associated adverse events such as gastrointestinal symptoms. Regarding the calcium concentration of dialysate (D-Ca) in Japan, 3.0 mEq/L is more popular than 2.5 mEq/L. Calcium loaded through 3.0 mEq/L dialysate may lead to metastatic calcification rather than to bone formation because serum phosphate concentration rebounds several hours after the end of each hemodialysis session when plasma pH is still high. In contrast, use of 2.5 mEq/L dialysate may result in an unfavorable increase of intact parathyroid hormone particularly when the amount of oral calcium intake is reduced. Although a higher dose of vitamin D is required to counteract the stimulation of parathyroid glands, hypercalcemia is less likely with 2.5 mEq/L dialysate. As the new K/DOQI guidelines are released, it is time to discuss the appropriate D-Ca as well as doses and kinds of phosphate binders and vitamin D for the comprehensive management of Japanese hemodialysis patients. PMID- 15828904 TI - Control of parathyroid function in patients with a short history of hemodialysis. AB - Secondary hyperparathyroidism (SHPT) is one of the serious complications in patients with chronic kidney disease. Parathyroid glands secrete parathyroid hormone (PTH), stimulated partly by hyperphosphatemia and hypocalcemia complicating chronic kidney disease (CKD). Use of a calcium-based phosphate binder might be sufficient to reduce serum PTH levels in mild SHPT, while the recent K/DOQI clinical guidelines recommended vitamin D therapy for dialysis patients with serum level of intact parathyroid hormone of 300 pg/mL or more. We conducted a 6-month prospective controlled trial of 50 patients initiating hemodialysis therapy who were randomized to receive oral calcium carbonate alone or the drug plus oral vitamin D sterol, calcitriol or alfacalcidol. The primary end point was the proportion of randomized patients who had a mean PTH level of 300 pg/mL or less at the end of this study. The secondary end point included the percent change in the values for corrected calcium, phosphorus, the calcium phosphorus product, and PTH. Eighty percent of patients receiving calcium carbonate without vitamin D sterols (20 of 25) reached the primary end point-a mean PTH level of 300 pg/mL or less after the period-as compared with 84% of those receiving calcium carbonate and vitamin D sterol (21 of 25) (Mantel Haenszel odds ratio 0.76, 95% confidence interval: 0.18-3.25, P = 0.71). The other effects of the two regimens on the secondary end points were not significantly different after 6 months. In SHPT of dialysis patients initiating hemodialysis, oral calcium carbonate use alone was not inferior to additional vitamin D sterol use with calcium carbonate in reducing serum PTH levels. Our result indicated that, if serum calcium and phosphorus levels are controlled primarily regardless of used agents, it will be followed by reduction of serum PTH level in these patients. PMID- 15828905 TI - Surgical indications and procedures of parathyroidectomy in patients with chronic kidney disease. AB - The Kidney Foundation of the USA proposed clinical practice guidelines for bone metabolism and disease in chronic kidney disease including parathyroidectomy (PTx). We performed PTx in a total of 1725 patients with advanced secondary hyperparathyroidism (2HPT) and evaluated the K/DOQI guideline concerning PTx, comparing it with our surgical strategy. The guidelines emphasize the avoidance of ectopic calcification and cardiovascular complications which may be induced by hypercalcemia, hyperphosphatemia, and persistent high parathyroid hormone level. We agree with the attitude of the K/DOQI guideline. To decide surgical indications, we emphasize that the size of parathyroid gland is one of the important factors. The guideline recommends subtotal PTx and total PTx with autotransplantation. We prefer total PTx with forearm autograft, mainly because it is easier and safer to remove the residual parathyroid tissue from the forearm at recurrence compared to neck re-exploration. In Japan, almost all patients require long-term hemodialysis after PTx because of the very small opportunity of kidney transplantation. The risk of recurrence is not negligible. Based on our huge experience we believe our strategy for advanced secondary hyperparathyroidism can be accepted at least in Japan. PMID- 15828906 TI - Efficacy of percutaneous ethanol injection therapy for secondary hyperparathyroidism in patients on hemodialysis as evaluated by parathyroid hormone levels according to K/DOQI guidelines. AB - Secondary hyperparathyroidism (SHPT) is a major complication of hemodialysis patients. Recently, percutaneous ethanol injection therapy (PEIT) has become a useful alternative treatment to parathyroidectomy (PTx). In this study, we evaluate the usefulness of PEIT for SHPT according to Kidney Disease Outcomes Quality Initiative (K/DOQI) guidelines. We studied 28 patients on hemodialysis with high intact-PTH (>400 pg/mL) and one to four swollen parathyroid glands detected by power Doppler ultrasonography. They were classified into Group 1 (N = 16), with 1 or 2 swollen glands, Group 2 (N = 5), with 3 or 4 swollen glands, and Group 3 (N = 7), high-risk patients for PTx. We compared serum intact-PTH levels 1 year after PEIT according to K/DOQI guidelines among these groups. We also evaluated the effectiveness of PEIT and PTx by comparing intact-PTH levels in 21 patients 1 year after PEIT (groups 1 and 2) with 11 patients after PTx. In Group 1, adequate intact-PTH levels were noted in 13 of 16 (81.2%) patients after PEIT, while 1 patient of 5 (20%) was achieved in Group 2, and 2 of 7 (28.6%) patients of Group 3. Adequate intact-PTH levels were attained in 14 of 21 (66.7%) patients of the PEIT group but only in 2 of 11 (18.2%) patients of the PTx group. Our results suggest that PEIT is a useful treatment for SHPT, especially in patients with one or two swollen glands. Through appropriate selection of patients for PEIT and correct injection of ethanol into the enlarged parathyroid gland, PEIT could accomplish better outcomes based on K/DOQI guidelines. PMID- 15828907 TI - LDL-apheresis accelerates the recovery of nonarteritic acute anterior ischemic optic neuropathy. AB - Nonarteritic acute anterior ischemic optic neuropathy (NAION) is a disabling disease which impairs visual function. It is presumed to result from disturbances of microcirculation in the anterior portion of the optic nerve head due to hemodynamic factors derived from excessive blood viscosity, or restriction of the vasal lumen in hypertensive, hypercholesterolemic, diabetic patients. We aimed to determine whether acute reduction of plasma fibrinogen and serum low-density lipoprotein (LDL) cholesterol is effective for treatment of NAION. We recruited 11 patients (7 females, 4 males) with a mean age of 57.2 +/- 19.6 years. All except one of them presented risk factors for atherosclerosis. The mean values of LDL-cholesterol and fibrinogen before treatment were 144 +/- 32 mg/dL and 341 +/- 80 mg/dL, respectively. All were treated with standard therapy (prednisone, salicylate, pentoxiphyllin) and underwent three sessions of LDL-apheresis (HELP system-B Braun) that can reduce plasma LDL-cholesterol and fibrinogen by more than 50% in a very short time. In all patients we observed a drastic reduction of LDL cholesterol and fibrinogen and a clear improvement in the visual functional data. In fact, mean values of corrected vision increased from 3.7/10 +/- 3/10 to 7.9/10 +/- 2.2/10 (P = 0.002) after the third session, while the scotomatous portion of the visual field regressed after the first session, and in 5 patients further regressed after the third session. This improvement had remained stable after 3 months. Thanks to it's effect of antagonizing hemorheologic disorders of the ocular microcirculation, fibrinogen/LDL-apheresis seems to be an efficacious treatment of NAION. PMID- 15828908 TI - Prevention of limb amputation in patients with limbs ulcers by autologous peripheral blood mononuclear cell implantation. AB - There are many cases of amputation of ischemic limbs of dialysis patients due to diabetes, despite the availability of medicine therapy and vascular by-pass operations. As there is extensive ruin of the vascular bed due to diabetes, vascular regeneration therapy by stem cell implantation is effective. Thirty patients with ischemic limbs due to diabetes (not including type-I) and on dialysis for chronic renal failure (19 cases), diabetes (5 cases), dialysis patients without diabetes (4 cases), and arteriosclerosis obliterans (ASO, 2 cases) were treated by autologous peripheral blood stem cell (PBSC) implantation where imminent amputation was under consideration. Granulocyte Colony Stimulate Factor (G-CSF: 5 microg/kg/day) was administered subcutaneously for 4 days before PBSC collection, that was carried out using a centrifuge (Spectra and/or CS3000) via the vein. The collected PBSC, containing 4.2 x 10(7) of CD 34 positive cells, was divided into units of 0.5-1.0 mL and implanted, without any purification, to the ischemic area of the limbs in about 65 points. In 21 cases, normalization of limb temperature was observed by thermograph, and symptoms also improved. The result of this first attempt of PBSC implantation is that we were able to save 22 ischemic limbs. This is the first large report of the application of regenerative medicine to peripheral ischemic limbs. PMID- 15828909 TI - Study of plasma levels of soluble CD40 ligand in systemic lupus erythematosus patients who have undergone plasmapheresis. AB - We studied whether soluble CD40 ligands (sCD40L) are removed by means of double filtration plasmapheresis (DFPP), and the removal may help decrease activity of systemic lupus erythematosus (SLE). We studied 10 female patients with active SLE. Double filtration plasmapheresis was conducted one or two times per week. Plasma sCD40L levels were measured before and after each round of DFPP and throughout the treatment course. The plasma sCD40L level of SLE patients was significantly higher (14.09 +/- 18.88 ng/mL) than that of healthy individuals (0.19 +/- 0.20 ng/mL; P < 0.0001). In the SLE patients, plasma sCD40L levels were significantly lower following DFPP (P = 0.0251). The plasma waste from DFPP of an SLE patient was subjected to gel filtration, and the sCD40L concentration in each fraction was measured. We observed a peak in the fraction corresponding to > or =60 kDa. These results indicate that trimers and higher order complexes of sCD40L are removed during DFPP. Plasma sCD40L level and SLE disease activity index (SLEDAI) were decreased following the treatment course (mean 9.3 months). sCD40L exists as both a monomer and trimer in the plasma of SLE patients. The trimer as well as higher-order compounds can be removed via DFPP. It was thought that removal of sCD40L via DFPP may be useful for improving the overall condition of SLE. PMID- 15828910 TI - Reversible leukoencephalopathy syndrome associated to leukopenia in a chronic hemodialysis patient. AB - Reversible leukoencephalopathy syndrome (RLS) is a rare brain disorder, characterized by diffuse attenuation of cerebral white matter, which has been most commonly observed in transplant patients receiving calcineurin inhibitors or in patients with severe hypertension. We report an episode of RLS in a 22-year old male patient on chronic hemodialysis with well-controlled moderate hypertension who presented with de novo headache and generalized seizures. Cranial magnetic resonance image (MRI) revealed multiple areas of increased signal intensity in the white matter on T2-weighed images which resolved spontaneously at subsequent MRIs. White blood cell count showed leucopenia with normal CD4 count at flow cytometry. A viral etiology could not be demonstrated. Reversible leukoencephaolopathy syndrome symptoms remitted within 72 h but leukopenia persisted over 10 months. The patient received a kidney transplant 15 months after RLS onset and has received cyclosporine since the second post transplant day. No recurrence of RLS symptoms has been observed. The etiology of the MRI changes in the present case seemed not to be either vasogenic or cytotoxic. PMID- 15828911 TI - Symptomatic bradycardia probably due to tizanidine hydrochloride in a chronic hemodialysis patient. AB - A 71-year-old woman was admitted to the Wakayama Medical University Hospital with dizziness and loss of body balance. She had started hemodialysis at the age of 70. During the 33 days before admission, she received oral tizanidine hydrochloride at 3 mg/day for leg cramps. An admission electrocardiogram (ECG) demonstrated sinus bradycardia of 47 bpm. A 24-h ECG showed a total number of heartbeats of 68,779 and an average heart rate of 48 bpm. The maximum RR interval was 3720 msec. The electrophysiology test demonstrated slight sinus node dysfunction. There was no major organic heart disease. We suspected that tizanidine was the cause of bradycardia and stopped administration of this drug. After discontinuation symptoms gradually disappeared. The serum concentration of the tizanidine showed a higher trough of 1.78 ng/mL. In conclusion, because there was a disappearance of symptoms and a lightening of bradycardia due to the discontinuation of this medication, tizanidine was strongly suspected as the cause of severe bradycardia. PMID- 15828913 TI - Letter to the editor. PMID- 15828923 TI - Suppression of inflammatory cytokine secretion by granulocyte/monocyte adsorptive apheresis in active ulcerative colitis. AB - To elucidate the molecular mechanisms involved in the therapeutic effects of granulocyte/monocyte adsorption apheresis, changes were investigated in the cytokine responses of peripheral blood mononuclear cells (PBMC) before and after granulocyte/monocyte adsorptive apheresis in ulcerative colitis (UC) patients. Four patients with active UC were enrolled. All patients responded to granulocyte/monocyte adsorptive apheresis. A total of 20 sessions of four patients were analyzed. Peripheral blood mononuclear cells were isolated from peripheral venous blood within 5 min before and after each session of granulocyte/monocyte adsorptive apheresis. The cells were stimulated with interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha for 24 h, and the secreted IL-8 and IL-6 levels were determined by enzyme-linked immunosorbent assay (ELISA). IL-1beta-induced IL-8 and IL-6 secretion was significantly decreased after granulocyte/monocyte adsorptive apheresis. TNF-alpha-induced IL-8 secretion was also significantly decreased after apheresis, but there was no significant difference in TNF-alpha-induced IL-6 secretion (P = 0.052). In conclusion, granulocyte/monocyte adsorptive apheresis down-regulates the IL-1beta and TNF-alpha-induced inflammatory responses in PBMC. The induction of hyporesponsiveness to pro-inflammatory cytokines may be an important factor mediating the clinical effects of granulocyte/macrophage adsorptive apheresis in UC patients. PMID- 15828924 TI - Effect over time of endotoxin adsorption therapy in sepsis. AB - Despite the use of potent antibiotics and intensive supportive care, the mortality among patients with sepsis and Gram-negative bacteremia remains high. In recent years, endotoxin adsorption therapy (PMX-DHP, polymyxin-direct hemoperfusion) has been widely used in Japan to remove endotoxin, a causative agent of sepsis. In septic patients whose clinical condition may change at any moment, the decision of when to perform blood purification in addition to conventional intensive care is a critical factor in the therapeutic strategy and prognosis. In the present study, we investigated the effect over time of PMX-DHP in sepsis. The subjects were 16 patients with systemic inflammatory response syndrome (SIRS) who required surgical treatment including a surgical operation and drainage. The following six parameters were compared between the first and second PMX-DHP: mean blood pressure and time-restricted urine at four time points - at baseline and at 6, 24 and 72 h after PMX-DHP; and white blood cell count, platelet count, base excess and Septic Severity Score (SSS) at 24 and 72 h after PMX-DHP. Mean blood pressure improved over time up to 24 h after both the first and second PMX-DHP. Time-restricted urine volume improved only at 6 h after the first PMX-DHP. White blood cell count improved over time up to 24 h after both the first and second PMX-DHP. The SSS improved at all time points studied except for 3 days after the second PMX-DHP. We conclude that PMX-DHP is expected to have important implications in terms of (i) correction of clinical conditions (by severity assessment); (ii) improvement of hemodynamics; (iii) possible anti inflammatory effect; and (iv) possible improvement of oxygen metabolism in tissues. PMID- 15828925 TI - Granulocyte transfusions in children with chronic granulomatous disease and invasive aspergillosis. AB - The transfusion of granulocytes to restore host defenses in severely granulocytopenic patients or in patients with defective granulocyte functions has been studied for more than 60 years. However, inadequate dosage of cells and inconsistent efficacy has limited the usage of these transfusions. Recently, the use of mobilizing agents such as granulocyte colony stimulating factors and dexamethasone has renewed interest in these treatment modalities. The present study is conducted to determine an appropriate method of enriched granulocyte collection with Fresenius AS.TEC.204 cell separator (Fresenius, Bad Homburg, Germany) and to evaluate the preliminary clinical results of granulocyte transfusion therapy in patients with chronic granulomatous disease and invasive Aspergillosis in parallel with in vitro granulocyte function. Three patients who have been treated for chronic granulomatous disease and invasive Aspergillosis received a total of 20 granulocyte transfusions. To mobilize granulocytes, healthy donors were given 450 microg of granulocyte colony-stimulating factor (G CSF) subcutaneously and 8 mg of dexamethasone orally approximately 12 h before collection. Five microg/kg/day of G-CSF was also subcutaneously administered prior to granulocyte transfusions. The first patient received 4; the second, 14 and the third, 2 transfusions. The granulocyte count given to these patients ranged between 0.4 and 3.0 x 10(9)/kg. Most transfusions were well tolerated. The nitroblue tetrazolium (NBT) tests that were done 16-24 h after the transfusion showed 14-46% dye reduction. Two of the three patients survived the infection. Granulocyte transfusions from G-CSF and dexamethasone stimulated donors could be a choice of treatment in chronic granulomatous disease patients, especially with disseminated invasive Aspergillosis. PMID- 15828926 TI - Is lipoprotein (a)-apheresis useful? AB - Numerous epidemiological investigations have shown the importance of cholesterol, and in particular low density lipoprotein (LDL), and of the lipoproteins in the development of coronary sclerosis. A continuing relationship between cholesterol levels and coronary morbidity has been established. The LDL concentration in the blood is, in particular, to be made responsible for the development of arteriosclerosis and especially of coronary heart disease (CHD). Lipoprotein (a) [Lp(a)], as a risk factor for premature cardiovascular and cerebrovascular diseases, can be lowered by LDL-apheresis. Especially in isolated high levels of Lp(a) with CHD or polygenic hypercholesterolemia with elevated Lp(a) levels, LDL apheresis can be indicated and can be useful to improve endothelium regulation and induce changes in coronary tone by an increase in endothelial derived relaxing factor. Lipoprotein (a) can be dramatically lowered by LDL-apheresis, but clinical improvement especially by low LDL is not still not clarified. Studies with weekly apheresis with statins versus drug therapy alone are necessary. To clarify the controversial discussions of whether lowering Lp(a) may be unnecessary or necessary to arrest progression of CHD, more clinical and randomized studies are needed. Lipoprotein (a) can be also lowered by current LDL apheresis methods. PMID- 15828927 TI - Autoimmune-associated congenital heart block: treatment of the mother with immunoadsorption. AB - Autoimmune-associated congenital heart block (CHB) is a rare complication of pregnancy in mothers with Anti-Ro/SSA antibodies (SSA-abs), resulting in fetal myocarditis, atrioventricular block, hydrops fetalis and/or intrauterine fetal death. As these antibodies are supposed to be directly involved in the pathogenesis of CHB, their removal should be associated with an improved clinical course. Extracorporeal immunoadsorption (IA) is the most efficient method to remove IgG-immunoglobulins like SSA-abs selectively. Two women with high titers of those auto-antibodies [mothers serum 615 and 612, respectively (normal range <3.0 IU/mL)] were treated with IA two to three times per gestation week in the outpatient department of the University of Rostock. In both patients, the mean removal of IgG (65 +/- 6%) to a target near 2.0 g/L after IA was successful. The SSA-abs were reduced from mean 328 +/- 138 and 247 +/- 105 pre IA to 88 +/- 124 and 98 +/- 42 post IA, respectively. One child received a pacemaker due to the persisting atrioventricular block grade III after birth. The second was unaffected. The removal of highly elevated SSA-antibodies by immunoadsorption is a possible treatment option in pregnant woman with high titers of those antibodies and/or a positive history of clinical complications. Further clinical studies are necessary. PMID- 15828928 TI - Non-invasive monitoring of hemodynamic changes during hemodialysis by the use of a newly developed admittance cardiograph. AB - Only a little information is available for the evaluation of the complex hemodynamic changes that occur during hemodialysis. Recently, we developed the transthoracic electrical admittance cardiograph for repeated measurements of cardiac output, and monitored hemodynamic changes during hemodialysis by the use of this device. We measured cardiovascular hemodynamic and autonomic parameters non-invasively during 210 min of hemodialysis in 19 chronic hemodialysis patients who for more than 2 months had no history of cardiovascular collapses during hemodialysis. Blood pressure was monitored every 10 min using a cuff oscillometric device (TM-2425; A & D, Tokyo, Japan). Cardiac output was monitored on a beat-by-beat basis by a newly developed electrical admittance cardiograph (NICOVIEW PA1100; NEC, Tokyo, Japan). Electrocardiogram R-R intervals were also monitored by the TM-2425. Power spectral analysis of R-R intervals was performed to obtain the low-frequency (LF; 0.05-0.15 Hz) and the high-frequency (HF; 0.15 0.40 Hz) components based on an autoregressive model. Change in circulatory blood volume was also monitored by a CRIT-LINE (In-Line Diagnostics, Riverdale, UT, USA). Although blood volume declined significantly by 16.3 +/- 1.4% (mean +/- SE) during hemodialysis (P < 0.0001), mean blood pressure did not change significantly. Heart rate increased significantly from the initial values of 76.3 +/- 3.4-86.4 +/- 4.9 beats/min (P = 0.03). Cardiac output and stroke volume decreased significantly from 4.47 +/- 0.30 to 2.91 +/- 0.32 L/min (P < 0.0001), and from 57.0 +/- 3.7 to 33.9 +/- 3.1 mL (P < 0.0001), respectively. Total peripheral vascular resistance increased significantly from 1940 +/- 146 to 3117 +/- 280 dyne x s x cm(5) (P < 0.0001). The LF component did not show significant change, while the HF component decreased significantly (P = 0.007), and the LF/HF ratio increased significantly (P = 0.03). These results suggest that a reduction in parasympathetic nerve activity and sympathetic nerve activation and a marked increase in total peripheral vascular resistance are responsible for the maintenance of blood pressure during hemodialysis in chronic hemodialysis patients. The admittance cardiograph seems to be one of the best options for serial measurements of cardiac output. PMID- 15828929 TI - Incidence of peripheral arteriosclerotic complications of the lower extremities in diabetic patients with chronic renal failure. AB - Patients with diabetes mellitus are at high risk of arteriosclerotic complications. The same is true for those with chronic renal failure (CRF). The present study evaluated clinical factors on the occurrence of peripheral arterial diseases. The severity of peripheral arterial disease was defined as mild (plaque, calcifications) or severe (70% stenosis, obstructions) based upon ultrasonographical examination. Overall in diabetic patients, mild ultrasonographical findings such as plaque and calcification were observed in 25% (17/69) of predialysis patients and in 18% (7/38) of those on hemodialysis (HD). Severe arteriosclerotic findings such as stenosis and obstruction were seen in 42% (29/69) of predialysis patients and in 50% (19/38) of those on HD. The incidence is identical between predialysis patients and patients on HD, regardless of the severity of peripheral arterial diseases. In an attempt to compare diabetics with non-diabetics, the incidence of mild abnormal findings was found in 22% (24/107) of diabetics and in 30% (11/37) of non-diabetics. Similarly, the percentage of patients with severe arteriosclerotic findings was found in 45% (48/107) in diabetics and 11% (4/37) in non-diabetics. The incidence of the severe type findings in the diabetics is significantly more frequent than that in the non-diabetics (P < 0.01, by the chi(2) test). In non-diabetic predialysis patients, the incidence of mild arteriosclerotic findings was observed in 30% (7/23), and 29% (4/14) of non-diabetic patients on HD. Similarly, the percentage of severe type was 9% (2/23) in non-diabetic predialysis patients, and 14% (2/14) in non-diabetic patients on HD. These data are not only supportive of previous data that diabetes is a risk factor for arteriosclerosis, but also suggestive that, even before the initiation of dialysis, patients with CRF are already susceptible to arteriosclerotic assault of peripheral arterial complications. PMID- 15828930 TI - Mathematical analysis of urea rebound in long-term hemodialysis patients. AB - Quantifying hemodialysis (HD) treatment requires knowledge of the equilibrated concentrations of the post-HD small molecule rebounds. However, measurement of the equilibrated concentrations is only possible after resting in bed after HD for at least 30 min, and this is often impractical. Therefore, we have analyzed mathematically the time course of post-HD urea rebound, and from this, have derived a new formula for predicting its equilibrated concentration. The blood urea nitrogen (BUN) was measured at 10 time points (immediately following HD, and 0.5, 2.5, 5, 7.5, 10, 15, 20, 25, and 30 min post-HD) in 12 anuric HD patients. The absolute change in the urea rebound (DeltaeqBUN) was approximated (DeltaestBUN) using the equation: DeltaestBUN = b -[1-exp x (-c x time (min))] + a x time (min). After the good correlation between DeltaeqBUN and DeltaestBUN, we compared the value of DeltaeqBUN measured at 30 min (DeltaeqBUN(30)) with that calculated (DeltaestBUN(30)) using only four sample points (immediately following HD, and 2.5, 5 and 10 min post-HD). Based on this result, we tried to predict post-HD BUN at 30 min (estBUN(30)). This study was undertaken to determine whether estBUN(30) may be representative of the equilibrated BUN (eqBUN(30)), and to compare with Kt/V using estBUN(30) and eqBUN(30). There was a significant correlation between DeltaeqBUN and DeltaestBUN (0.97 < r < 0.99, P < 0.001). Thus, there was a significant positive linear correlation between eqBUN(30) and estBUN(30) (eqBUN(30): 25.7 +/- 2.25 mg/dL, estBUN(30): 26.3 +/- 2.31 mg/dL; r(2) = 0.99, P < 0.001). A Kt/V measurement was obtained with single pool model using BUN just after HD (Kt/V(sp)), eqBUN(30) (Kt/V(eq)), and estBUN(30) (Kt/V(est)), and with double pool model using Kt/V(sp) (Kt/V(dp)) and was compared with them. Though Kt/V(sp) was significantly higher than Kt/V(eq) (1.26 +/- 0.08 vs. 1.09 +/ 0.07, P < 0.001), there were no differences among Kt/V(eq), Kt/V(est) and Kt/V(dp) (Kt/V(est): 1.06 +/- 0.07, Kt/V(dp): 1.10 +/- 0.07) and all values were clinically acceptable. Furthermore, there was a significant positive linear correlation between Kt/V(eq) and Kt/V(est) (r(2) = 0.98, P < 0.001). In conclusion, we have devised the method to predict equilibrated BUN and calculate double pool Kt/V, which requires samples up to 10 min post-HD. PMID- 15828931 TI - Effects of glucose and plasminogen activator inhibitor-1 on collagen metabolism in the peritoneum. AB - Nonphysiological solutions containing high glucose levels have been considered an important factor in the etiology of fibrotic changes in long-term continuous ambulatory peritoneal dialysis (CAPD) patients. At the same time, increased Plasminogen Activator Inhibitor (PAI)-1 secretion has been reported to correlate with fibrotic changes. We suspected that the high glucose content of peritoneal dialysis solution may induce peritoneal sclerosis via up-regulation of PAI-1 gene expression. In this study, we evaluated the effects of glucose on PAI-1 activity in peritoneal fibrosis in a rat model of CAPD. The effects of glucose on the expressions of PAI-1 and several other genes correlated with collagen metabolism were also examined in cultured rat peritoneal mesothelial cells and fibroblasts. Sprague-Dawley rats were intraperitoneally injected twice daily for 28 days with phosphate-buffered saline (PBS) (control group), PBS containing 4% glucose (glucose group), or PBS containing 4% glucose plus a PAI-1 inhibitor (PAI-1 inhibitor group). Thickening of the peritoneum with increase the deposition of collagens type I and III in the submesothelial interstitium were observed in the glucose and the PAI-1 inhibitor group, but these were less severe in the PAI-1 inhibitor group. Glucose stimulated expression of the mRNA of PAI-1, collagen type I and III, and tissue inhibitor of metalloproteinase (TIMP)-1 in fibroblasts but not in mesothelial cells. Glucose stimulated matrix metalloproteinase (MMP) 13 mRNA expression in both cell types. The PAI-1 inhibitor suppressed expression of the mRNAs induced by glucose. In conclusion, glucose induces peritoneal fibrosis, including changes in collagen metabolism, by stimulating PAI-1 expression. PMID- 15828932 TI - Synergistic cytotoxicity of acidity and 3,4-Dideoxyglucosone-3-ene under the existence of lactate in peritoneal dialysis fluid. AB - Of the non-physiological compounds in glucose-rich peritoneal dialysis fluid, we investigated the synergistic cytotoxicity of acidity and 3,4-Dideoxyglucosone-3 ene(3,4-DGE) under the existence of lactate using human peritoneal mesothelial cells (HPMC). The effect of pH on cell viability at various levels of pH (5.5, 6.7, 7.15), with or without lactate was examined by adding 1N-HCl to phosphate buffer solution. We also examined the cytotoxic effects of 3,4-DGE and pH (5.5, 6.7 or 7.15). Additionally, we compared the cytotoxic effects of 3,4-DGE and pH (5.5, 6.7 or 7.15) under existence of lactate (40 meq/L) or absence of lactate. The cells were exposed to these solutions for 2 or 4 h. Cell viability was determined by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenylterazolium bromide) assay. 3,4-DGE or acidic solution alone had no significant effects on MTT viability under the absence of lactate. However, acidic solutions containing 3,4 DGE significantly decreased MTT viability under the existence of lactate. The MTT viability of HPMC was not decreased by 3,4-DGE or acidity alone under the absence of lactate. However, the combination of acidity and 3,4-DGE markedly decreased MTT viability under the existence of lactate, strongly suggesting the synergistic cytotoxicity of 3,4-DGE and acidity under the existence of lactate. PMID- 15828936 TI - Engineered growth factors and cutaneous wound healing: success and possible questions in the past 10 years. AB - In the past 10 years, many engineered growth factors, including recombinant human epidermal growth factor, basic fibroblast growth factor, and platelet-derived growth factor, have been produced and used in the clinic. After screening the results from different centers, some results are found to be encouraging, while others are discouraging. Although the interpretation of these results may depend on your perspective, it may also depend on different criteria, different wounds, and even different aims. In this article, successful experiences and failures concerning the use of growth factors and cutaneous wound healing are summarized. Based on this information and our clinical experience, we address people's concerns such as whether growth factors have altered clinical practice thus far and whether growth factor treatments have solved all problems involved in wound healing. Is there a need for exogenous application of growth factors in acute or chronic wounds, and if so, is it safe to use growth factors to promote wound healing? Last, can we achieve perfect wound healing in those wounds treated with growth factors? PMID- 15828937 TI - Effect of sharp debridement using curette on recalcitrant nonhealing venous leg ulcers: a concurrently controlled, prospective cohort study. AB - The objective of this study was to evaluate the effect of sharp debridement on the progression of recalcitrant chronic venous leg ulcers (CVLU) and to assess the feasibility of performing this procedure in an outpatient setting. We performed a prospective study of 55 CVLU (53 patients) over a 12-month period. The study group, which underwent debridement, contained 28 CVLU whose wound beds had slough, nonviable tissue, and no granulation tissue. The control group was 27 CVLU with minimal (15-20%) granulation tissue, but no slough or nonviable tissue. Treatments were otherwise similar. Age, body mass index, mean ulcer surface area (MSA) and mean ulcer duration were comparable in both groups. Ulcer measurements were taken at 4 weeks before debridement, at the time of debridement, and 4 and 20 weeks post-debridement. There was no change in the MSA from 4 weeks before to the time of debridement in either group. At 4 weeks post-debridement, the study ulcers showed a 6 cm(2) reduction in the MSA vs. a 1 cm(2) reduction in controls (P = 0.02). By week 20 post-debridement, the study ulcers achieved a 7.4 cm(2) reduction in the MSA vs. an increase of 1.3 cm(2) in controls (P = 0.008). Between weeks 8 and 20 post-debridement, 16% of study ulcers vs. 4.3% of control ulcers achieved complete healing. Infection rates and antimicrobial usage were similar. We conclude that sharp debridement is effective in stimulating healing of recalcitrant CVLU. It is safe, well tolerated, and can be performed in an outpatient setting. PMID- 15828939 TI - Multiple fragments related to angiostatin and endostatin in fluid from venous leg ulcers. AB - To investigate whether compromised angiogenesis could contribute to the impaired healing of venous leg ulcers, we have analyzed fluids from venous leg ulcers for the presence of the angiogenesis inhibitors angiostatin and endostatin. Multiple fragments related to angiostatin were detected by Western blot analysis. One angiostatin fragment was identified by mass spectrometry as plasminogen kringle domains 1-3 containing amino acids 82-343 of plasminogen, and a fraction containing this fragment inhibited tubule formation of human umbilical vein endothelial cells in a Matrigel assay. The leg ulcer fluids also contained endogenous endostatin (20 kDa) as well as higher molecular weight endostatin related proteins. The concentrations of endostatin in the wound fluids, which ranged from 12.8 to 65.5 ng/ml, were higher than the concentration in human serum (7.7 ng/ml). Most of the endostatin in leg ulcer fluid appeared to be bound to the proteoglycan glypican-1. These data suggest that anti-angiogenic activity is present at the site of venous leg ulcers, and at least in the case of angiostatin, is biologically active. PMID- 15828938 TI - Efficacy and safety of the freeze-dried cultured human keratinocyte lysate, LyphoDerm 0.9%, in the treatment of hard-to-heal venous leg ulcers. AB - LyphoDerm (XCELLentis, Belgium) is an end-sterilized, freeze-dried lysate from cultured allogeneic epidermal keratinocytes, formulated into a hydrophilic gel. Its efficacy and safety were evaluated, in combination with standard care (hydrocolloid dressing and compression therapy), in 194 patients suffering from hard-to-heal (lasting more than 6 weeks and not responding to conventional therapy) venous leg ulcers. Two control groups received standard care, with or without vehicle, respectively. Patients had a median age of 67.5 years and the majority were females (61%). The median duration of the ulcer was 43 weeks and in 39% of the subjects it had been present for more than 1 year. Thirty-eight percent of the patients in the standard care + LyphoDerm group had complete ulcer healing within 24 weeks (primary end point) compared to 27% of patients in the standard care + vehicle pooled groups (P = 0.114) in the "as treated" intent-to treat cohort (37% vs. 27% in the "as randomized intent-to-treat cohort; p = 0.137). In the subgroup of patients with enlarging ulcers, the difference between the two groups was significant (30% vs. 11%; p = 0.024 in the "as treated" intent to-treat cohort and 31% vs. 9%; p = 0.005 in the "as randomized" intent-to-treat cohort). LyphoDerm was well tolerated and safe, and no differences in the frequency of adverse events were noted between the treatment groups. Although the primary objective of the study was not achieved, the exploratory analysis carried out in patients with enlarging ulcers suggests that LyphoDerm could offer a new prospect for the treatment of patients with venous ulcers that may prove to be a significant adjunct to the overall provision of care. PMID- 15828940 TI - Hypoxia increases reepithelialization via an alphavbeta6-dependent pathway. AB - Reepithelialization is an essential step in successful cutaneous wound healing. Human keratinocytes, integral in this process, have been shown to have increased motility in the hypoxic healing edge of wounds correlating with the clinical success of semiocclusive hypoxic dressings, although the underlying mechanisms remain poorly understood. Subconfluent human keratinocyte cell monolayers were exposed to 1% hypoxia for up to 24 hours or control conditions. Re-oxygenation studies were performed up to 72 hours. Cellular alphav subunit and alphavbeta6 integrin expression was measured by flow cytometry. Migration scratch assays on fibronectin following hypoxic exposure were performed over 24 hours. Relative matrix metallo-proteinase (MMP)-2, 9 activity was determined by gelatin zymography with TIMP-1 levels assayed by enzyme-linked immunoassay. Sustained increases in alphav and alphavbeta6 expression were shown up to 48 hours in re oxygenation studies (P < 0.001). Standardized scratch assays confirmed increased migration in the hypoxic group (P < 0.05). This effect was attenuated by the addition of a specific inhibitor of the alphavbeta6 integrin. MMP-2 and -9 activity was up-regulated following hypoxic exposure (P < 0.001; P < 0.05, respectively), whereas increased MMP expression was significantly retarded by addition of an alphavbeta6 inhibitor (P < 0.05). Migration on fibronectin was attenuated by a specific gelatinase inhibitor. We conclude that integrin alphavbeta6-dependent MMP-2 and -9 up-regulation is an important feature of increased migration in hypoxic human keratinocytes. PMID- 15828941 TI - Opioids heal ischemic wounds in the rat. AB - Opioids are sometimes used to treat pain in ulcerative wounds, and it is speculated that pain interferes with the healing process. Because the direct effect of opioids on this process remains unknown, we examined the effect of topically applied opioids on the healing of open ischemic wounds in rats. Topically applied opioids hastened wound closure, particularly in the first 4 days when no healing was initiated in phosphate buffered saline solution-treated wounds. After 1 week of application, fentanyl, hydromorphone, and morphine resulted in 66%, 55%, and 42% wound closure, respectively, as compared to only 15% in control wounds. Opioid-induced healing was accompanied by a 1.5- to 2.5 fold increase in nuclear density in the granulation tissue and 45-87% increase in angiogenesis as compared to phosphate buffered saline solution-treated wounds. Fentanyl showed significantly improved healing compared to morphine and hydromorphone (p < 0.05, fentanyl vs. others). Fentanyl-induced healing was inhibited by the opioid receptor antagonist naloxone, suggesting that peripheral opioid receptor(s) mediate the healing process. Opioids accelerate healing by up regulating both endothelial and inducible nitric oxide synthase and the vascular endothelial-derived growth factor receptor Flk1 in the wounds. We envision that opioids can be used topically to accelerate wound healing in diverse clinical conditions ranging from surgical incisions to nonhealing ischemic ulcers in pathophysiological conditions and in hospice patients. PMID- 15828942 TI - Wound healing in denervated rat skin. AB - Recently, several reports have suggested that innervation influences wound healing. However, some investigators have reported that nerve injury prevented wound healing while others have suggested it had no influence on full-thickness skin wound healing. We created denervated skin areas on rats by dissection of the spinal hemicord. Subsequently, 15-mm-diameter skin defects were made symmetrically within the denervated area on the right side of the back and the normal innervated area on the left side. Biopsies were performed at 3, 7, and 14 days after wounding. We measured changes of the wound surface area, the rate of wound contraction, and the rate of epithelialization. The differences were not significant at 3 or 7 days after the operation. However, we could observe significantly delayed wound healing of the denervated skin areas compared to the normal areas at 14 days. Both wound contraction and epithelialization were delayed in the denervated groups. Our results suggest that sensory disturbance is a negative factor for skin wound healing. PMID- 15828943 TI - Hyaluronic acid production and CD44 expression in cultured dermal fibroblasts of patients with non-insulin-dependent diabetes mellitus with and without chronic ulcers on the lower extremity. AB - It is well known that hyaluronic acid and its principal receptor, CD44, are implicated in the regulation of the tissue repair process, but their role in the formation of chronic diabetic ulcers has not been studied. Hyaluronic acid metabolism and CD44 expression are regulated by lactate, where their increased production is considered to affect the properties of fibroblasts in non-insulin dependent diabetes mellitus. The aim of our work was to investigate the possible role of hyaluronic acid and CD44, and their regulation by lactate, in the abnormal wound healing of diabetes. Fibroblasts were derived from uninjured skin from four non-insulin-dependent diabetic patients with ulcers and four without ulcers; and from four healthy age-matched volunteers. We observed that diabetic fibroblasts of both groups produced more L-lactate ( approximately 30%) and incorporated more (3)H-glucosamine into the medium hyaluronic acid ( approximately 28%) than controls. Fibroblasts of the diabetic group with ulcers, unlike those of the group without ulcers, showed significant increases in the high molecular weight hyaluronic acid accumulation in the pericellular matrix (30.5%, p < 0.01) and CD44 expression (27.0%, p < 0.05). Exogenous L-lactate dose dependently, and equally for all fibroblasts lines, stimulated the accumulation of medium hyaluronic acid (3.7-fold) and CD44 expression (1.5-fold). However, fibroblasts from diabetic patients with ulcers were more (1.4-fold) sensitive to L-lactate in terms of CD44 expression, and responded to L-lactate by the increased accumulation of high molecular weight hyaluronic acid in the pericellular matrix (32.1%, p < 0.01). We propose that specific properties of fibroblasts from diabetic patients with ulcers may be involved in the increased susceptibility of these patients to chronic ulceration. PMID- 15828944 TI - Deep dermal burn injury results in scarless wound healing in the ovine fetus. AB - Early to mid-term fetuses heal cutaneous incisional wounds without scars; however, fetal response to burn injury has not been ascertained. We present a fetal model of thermal injury and subsequent analysis of fetal and lamb response to burn injury. A reproducible deep dermal burn injury was created in the fetus by application of water at 66 degrees C for 7 seconds, and at 82 degrees C for 10 seconds to the lamb. Macroscopically, the area of fetal scald was undetectable from day 7 post injury, while all lamb scalds were readily identified and eventually healed with scarring. Using a five-point histopathology scoring system for alteration in tissue morphology, differences were detected between control and scalded skin at all stages in lamb postburn, but no difference was detected in the fetal model after day 7. There were also large differences in content of alpha-smooth muscle actin and transforming growth factor-beta1 between control and scalded lamb and these differences were statistically significant at day 14 (P < 0.01). This novel model of fetal and lamb response to deep dermal injury indicates that the fetus heals a deep burn injury in a scarless fashion. Further elucidation of this specific fetal process of burn injury repair may lead to improved outcome for patients with burn injury. PMID- 15828945 TI - Collagen in the scarless fetal skin wound: detection with picrosirius polarization. AB - Our group has developed an ovine model of deep dermal, partial-thickness burn where the fetus heals scarlessly and the lamb heals with scar. The comparison of collagen structure between these two different mechanisms of healing may elucidate the process of scarless wound healing. Picrosirius staining followed by polarized light microscopy was used to visualize collagen fibers, with digital capture and analysis. Collagen deposition increased with fetal age and the fibers became thicker, changing from green (type III collagen) to yellow/red (type I collagen). The ratio of type III collagen to type I was high in the fetus (166), whereas the lamb had a much lower ratio (0.2). After burn, the ratios of type III to type I collagen did not differ from those in control skin for either fetus or lamb. The fetal tissue maintained normal tissue architecture after burn while the lamb tissue showed irregular collagen organization. In conclusion, the type or amount of collagen does not alter significantly after injury. Tissue architecture differed between fetal and lamb tissue, suggesting that scar development is related to collagen cross-linking or arrangement. This study indicates that healing in the scarless fetal wound is representative of the normal fetal growth pattern, rather than a "response" to burn injury. PMID- 15828946 TI - MRL mice fail to heal the heart in response to ischemia-reperfusion injury. AB - The MRL/MpJ mouse strain has been reported to recover after right ventricular cryoinjury without scar formation or evidence of ventricular dysfunction, suggesting that this mouse strain harbors genetic traits that confer the capacity for adult myocardium to regenerate. We therefore sought to assess the capacity of adult MRL myocardium to regenerate in a left ventricular ischemia-reperfusion model of myocardial infarction, which more closely recapitulates injury that occurs in human disease. MRL (n = 13) and control C57/Bl6 (n = 12) mice underwent transient occlusion of the left anterior descending coronary artery. After 10 weeks, MRL and C57Bl/6 mice were euthanized and the extent of infarcted myocardium quantified with (2,3,5)-triphenyltetrazolium chloride and trichrome staining. There was no evidence of resistance to cardiac injury or of reduced scar formation in the MRL mice compared to C57/Bl6 controls. Myocardial infarct size (percentage of total heart weight +/- SEM) did not significantly differ between MRL and C57/Bl6 controls (18.9 +/- 1.8% for MRL vs. 15.7 + 1.3% for C57/Bl6, p = 0.20). Thickness of the infarcted anterior LV wall at the mid papillary level normalized to body weight was not significantly different between the two groups (0.017 + 0.003 mm/mg for MRL and 0.017 + 0.002 mm/mg for C57/BL6, p = 0.91). Trichrome staining showed intense scar formation in both C57/BL6 and MRL hearts. We conclude that there appears to be no effect of the MRL genetic background on resistance to myocardial infarction in mice. PMID- 15828947 TI - Analysis of ischemia-reperfusion injury in a microcirculatory model of pressure ulcers. AB - The aim of this study was to establish a pressure ulcer model that visualizes the microcirculation, and to examine the participation of ischemia-reperfusion injury in the pathophysiology of pressure ulcers. An original system composed of a new skin fold chamber and compression device allowed loading quantitative vertical stress to the skin. An intravital microscopic technique enabled direct visualization of the microcirculation in the physiological condition and in response to pressure application. To estimate the effect of ischemia-reperfusion injury, animals were divided into two groups: the compression-release group (n = 8), in which the animals received four cycles of compression-release which consisted of 2 hours of compression followed by 1 hour of pressure release; and the compression alone group (n = 8) in which the animals underwent continuous compression for 8 hours. Functional capillary density was quantified before the compression procedure and on day 1 (35 hours) after the first evaluation. The cyclic compression-release procedure significantly decreased functional capillary density as compared to continuous compression, indicating that in our experimental setting repetition of ischemia-reperfusion cycle more severely damaged the microcirculation than single prolonged ischemic insult. This finding supports the significant contribution of ischemia-reperfusion injury to the pathophysiology of pressure ulcers at the level of dynamic in vivo microcirculation. PMID- 15828949 TI - Presentation and therapy of myelolipoma. AB - BACKGROUND: Adrenal myelolipoma is a rare and benign, hormonally inactive tumor frequently discovered incidentally. Because of the increasing rate of detection of adrenal myelolipoma, use of the correct diagnostic examination and treatment, with respect to surgical excision or regular controls, is continually gaining importance. We report herein on the largest series of surgically treated patients with adrenal myelolipoma from a single institute. METHODS: The clinical charts of 12 patients who underwent adrenal surgery for myelolipoma were reviewed. Follow up was carried out by means of re-examination or by personal contact with the primary physician. It was possible to perform a re-examination on eight of the 12 patients 6.9 years after the operation: one patient had died 5.2 years after surgery. RESULTS: Diagnosis of adrenal myelolipoma was made incidentally in seven patients, and during examination for reasons of unspecific abdominal or flank pain in five patients. A transabdominal approach was taken in five patients, a subcostal approach was taken in three patients and a translumbal approach was taken in four patients. At the follow up, all formerly symptomatic patients were free of symptoms. No recurrence could be seen; however, in one patient a contralateral adrenal myelolipoma had developed. CONCLUSIONS: Symptomatic tumors, growing tumors or tumors larger than 10 cm should be excised surgically by means of an endoscopic or conventional approach, depending on the size. Surgical indication should not be liberalized by the introduction of more gentle operative techniques such as the translumbal or the endoscopic approach. A close follow up should be maintained in the case of patients free of symptoms, and for those with a definite diagnosis from imaging procedures. PMID- 15828950 TI - The relationship of magnesium intake to serum and urinary calcium and magnesium levels in Trinidadian stone formers. AB - BACKGROUND: The present study was undertaken to investigate the relationship between the dietary intake of magnesium and the serum and urinary levels of calcium and magnesium in a group of Trinidadian stone formers. METHODS: A group of 102 confirmed stone formers presenting to urological clinics were interviewed using a questionnaire designed to obtain a semi-quantitative estimate of their oral magnesium intake. Patients were invited to give blood samples for serum calcium and magnesium levels and to provide 24-h urine specimens for the measurement of urinary levels of these minerals, as well as total urinary volumes. A group of 102 controls was subjected to a similar interview and blood and urinary testing. Chi-square tests and Student's t-tests were used to examine group demographic differences. The Mann-Whitney test investigated differences in biochemical indices. Binary logistic regression was used to identify predictors of stone formation. RESULTS: Blood samples were obtained from 60 patients and 98 controls. Urine samples were returned by 34 patients and 97 controls. Only 10 stones were retrieved from patients. Patients had a significantly lower magnesium intake, but higher median serum and urinary calcium levels, and higher serum calcium to magnesium ratios than controls. Independent variables capable of predicting stone formation included total magnesium intake and serum and urinary calcium levels. CONCLUSIONS: Increased serum and urinary calcium levels, calcium to magnesium ratios, and a low magnesium intake were predictive of stone formation in this Trinidadian population. PMID- 15828951 TI - Prognostic significance of vascular invasion in patients with bladder cancer who underwent radical cystectomy. AB - BACKGROUND: The objective of this study was to determine whether vascular invasion (i.e. lymphatic and blood vessel invasion) could be a useful prognostic predictor in patients with locally invasive transitional cell carcinoma (TCC) of the bladder who underwent radical cystectomy. METHODS: This series included 114 consecutive patients undergoing radical cystectomy for primary TCC of the bladder between November 1989 and July 2003. Several clinicopathological characteristics of these patients were analyzed, focusing on the association between vascular invasion and disease recurrence after radical cystectomy. RESULTS: Lymphatic and blood vessel invasions were detected in 55 (48.2%) and 33 (29.8%) specimens, respectively. Lymphatic invasion was significantly associated with pathological stage, tumor grade, lymph node metastasis, blood vessel invasion and disease recurrence, whereas blood vessel invasion was significantly related to pathological stage, lymph node metastasis, lymphatic invasion and disease recurrence. Recurrence-free survival in patients with lymphatic invasion was significantly lower than that in those without lymphatic invasion, and a similar significant difference in recurrence-free survival was observed between patients with and without blood vessel invasion. However, multivariate analysis using the Cox proportional hazards model showed that only pathological stage and lymph node metastasis could be used as independent predictors for disease recurrence after radical cystectomy. CONCLUSIONS: Despite a significant association between several prognostic parameters, vascular invasion was not an independent predictor of disease recurrence; therefore, if there are other conventional parameters available, there might not be any additional advantage to considering the presence of vascular invasion when predicting the prognosis of patients undergoing radical cystectomy for TCC of the bladder. PMID- 15828952 TI - The long-term voiding function and sexual function after pelvic nerve-sparing radical surgery for rectal cancer. AB - BACKGROUND: The aim of the present study is to symptomatically analyze the extent to which pelvic nerve-sparing radical surgery for rectal cancer impacts on long term voiding and male sexual function. METHODS: A self-administered questionnaire was mailed to 68 patients who underwent pelvic nerve-sparing radical surgery for invasive rectal cancer with 52 responses (28 men and 24 women; 27 complete and 25 incomplete preservation; response rate 76.5%). Each patient was asked to record if there had been any changes in lower urinary tract symptoms after surgery. Sexual function was also investigated in men. RESULTS: Of the 52 patients, 48 (92%) maintained voluntary voiding without catheterization in the long term. Clean intermittent self-catheterization was performed in only four patients with incomplete preservation because of persistent voiding dysfunction. Subjectively, approximately 60% of the patients remained unchanged in lower urinary tract symptoms after surgery. The satisfaction rate regarding the current voiding status was significantly higher in women than in men (83% versus 61%, P = 0.0294), but was not significantly different between those with complete (76%) and incomplete preservation (64%). Despite the acceptable urinary status, 88% of men had some deterioration in the erectile function, regardless of the types of surgical procedures. Overall, 64% of men were unsatisfied with the current sexual function. CONCLUSIONS: Pelvic nerve-sparing radical surgery for rectal cancer preserved the long-term voiding function in the majority of patients. In completely preserved patients and in women, symptomatic outcomes were more satisfactory. Postoperative erectile dysfunction was found to be a serious problem, even in complete nerve-sparing procedure. PMID- 15828953 TI - Resistance index in benign prostatic hyperplasia using power Doppler imaging and clinical outcomes after transurethral vaporization of the prostate. AB - BACKGROUND: Using power Doppler ultrasonography (PDUS), we investigate the change of resistance index (RI) before and after transurethral vaporization of the prostate (TUVP) in benign prostatic hyperplasia (BPH) patients. METHODS: In all, 49 patients underwent transrectal PDUS before receiving TUVP, three were excluded because of cancer and three could not be followed up. The remaining 43 were enrolled in the present study. Patients were assessed before and 1, 3 and 6 months after surgery, giving a mean duration of follow-up of 9.1 months. International prostate symptom scores (IPSS), quality of life (QOL) scores, postvoiding residual urine volumes (PVR) and maximum urinary flow rates (Qmax) were evaluated and total prostatic volume (TPV) and RI were measured using PDUS. RESULTS: Resistance index ranged from 0.64 to 0.91. The postoperative parameters except for RI, such as TPV, PVR, IPSS and QOL scores improved significantly at the follow-up assessment after surgery. The elevated RI decreased significantly 1, 3 and 6 months after the treatment. Resistance index significantly decreased after TUVP and IPSS and other urodymamics parameters improved. CONCLUSIONS: The present study suggested that RI could evaluate the severity of BPH and the degree of intraprostatic pressure or bladder outlet obstruction. PMID- 15828954 TI - Prediction of potentially insignificant prostate cancer in men undergoing radical prostatectomy for clinically organ-confined disease. AB - BACKGROUND: The objectives of the present study were to characterize, according to tumor significance, the clinicopathological features of patients with prostate cancer who underwent radical prostatectomy, and to determine useful parameters for predicting insignificant disease before surgery. METHODS: In this series, we included 195 patients who underwent radical prostatectomy for clinically organ confined prostate cancer at our institution between January 1999 and November 2003. Several clinicopathological factors were analyzed, focusing on whether the largest tumor volume in radical prostatectomy specimens was >/=0.5 cm(3) or <0.5 cm(3), which is the criterion defined to distinguish insignificant cancer from significant cancer. RESULTS: Potentially insignificant cancer was detected in 28 of 195 patients (14.4%). There were significant differences between patients with insignificant disease and those with significant disease in serum prostate specific antigen (PSA) as well as all biopsy parameters, with the exception of biopsy Gleason score. Furthermore, final pathological examination demonstrated that these two patient groups showed significant differences in Gleason score and the incidence of extraprostatic disease extension. Pearson's correlation analysis showed that tumor volume in the prostatectomy specimens was significantly associated with serum PSA and all biopsy parameters; however, each of the correlations was comparatively weak. There was no single parameter that could viewed as a useful predictor of tumor significance. The best model for predicting insignificant tumor was the combined use of biopsy Gleason score <7 and percent of positive biopsy core (PPBC) <15%; however, the further addition of serum PSA <10 ng/mL to these two parameters failed to enhance the predictive value of cancer significance. CONCLUSIONS: These findings indicate that clinicopathological findings suggesting favorable features are well observed in patients with potentially insignificant prostate cancer compared to those with significant prostate cancer, and the combination of a biopsy Gleason score <7 and PPBC <15% might be useful as a predictor of insignificant disease. PMID- 15828955 TI - Intussusception of the bladder neck does not promote early restoration to urinary continence after non-nerve-sparing radical retropubic prostatectomy. AB - BACKGROUND: The objectives of the present study were to investigate whether buttressing sutures, which prevent the bladder neck from pulling open as the bladder fills, can promote earlier recovery from urinary incontinence after radical retropubic prostatectomy (RRP) and to identify possible risk factors associated with urinary incontinence after RRP. METHODS: The present study included 72 patients who underwent non-nerve-sparing RRP without neoadjuvant therapy between January and December 2003. Among these 72 patients, intussusception of the bladder neck was performed in 24 who consented to this procedure. In the present series, continence was defined as the absence of any need to use sanitary pads or diapers. Continence was evaluated by a patient interview 1, 3 and 6 months after RRP. RESULTS: There were no significant differences in clinicopathological characteristics between patients with and without intussusception of the bladder neck. The percentage of continent patients 1, 3 and 6 months after RRP was 34.7%, 63.9% and 95.8%, respectively, and there were no significant differences in continence between the two groups at any time point. Among several factors examined, only bladder neck preservation was an independent predictor of recovery from urinary incontinence 1 and 3 months after RRP. CONCLUSIONS: These findings suggest that it would be important to preserve the bladder neck for early return to continence after non-nerve-sparing RRP; however, intussusception of the bladder neck may not offer significant improvement in earlier return of urinary control. PMID- 15828956 TI - Scrotal dartos flap for the prevention of the urethrocutaneous fistula on hypospadias urethroplasty. AB - BACKGROUND: Urethrocutaneous fistula is a common complication of urethroplasty for severe hypospadias, even when a microsurgical technique is applied. PATIENTS AND METHODS: From June 2001 to July 2003, we applied the scrotal dartos flap wrapping technique to prevent the occurrence of urethrocutaneous fistula in 14 patients with hypospadias. RESULTS: Hypospadias was repaired without fistula formation in all but one patient, who developed a tiny fistula which was later closed successfully by a simple procedure. CONCLUSIONS: We recommend the scrotal dartos wrapping technique for covering the neourethra with a well-vascularized flap because of fewer complications, although the procedure to obtain the vascularized flap may take longer. PMID- 15828957 TI - Lectin-reactive alpha-fetoprotein as a marker for testicular tumor activity. AB - BACKGROUND: Lens culinaris agglutin (LCA)-affinity electrophoresis resolves serum alpha-fetoprotein (AFP) into three isoforms, AFP-L1, -L2 and -L3. The ratio of AFP-L3 to total AFP (AFP-L3%) is frequently high in hepatocellular carcinoma (HCC) patients, and thus, it is widely used for early diagnosis of HCC. In the present study, we used the subfraction profile of LCA-binding AFP to diagnose and monitor testicular tumor activity. METHODS: Serum samples were collected from 21 testicular tumor patients, and the LCA-reactive fractions were determined by LCA affinity electrophoresis coupled with antibody-affinity blotting. The histological diagnosis was non-seminomatous germ cell tumor (NSGCT) in 15 patients and pure seminoma in six patients. RESULTS: Serum AFP levels were abnormally elevated (>20 ng/mL) in 10 of 15 NSGCT patients. One NSGCT patient and two seminoma patients showed borderline AFP levels between 10 and 20 ng/mL. LCA reactive AFP was detected in all 11 NSGCT patients with serum AFP levels above 10 ng/mL, but not in the two seminoma patients with serum AFP levels above 10 ng/mL. In testicular tumor patients, the broad band of AFP-L2 could not be completely separated from AFP-L3. The mean ratio of AFP-L3 plus AFP-L2 (AFP-L2 + 3%) was as high as 94% (range 80-99%) in these patients. Serial determinations of LCA reactive fractions were performed in eight of the 11 LCA-reactive AFP-positive patients. They included five patients who received chemotherapy, and three patients who underwent orchiectomy for stage I NSGCT. In three of eight patients, LCA-reactive AFP was detected even after normalization of total AFP levels. All three patients relapsed, with elevation of serum AFP within several months. CONCLUSION: Determination of LCA-reactive AFP might be a useful marker for testicular tumor activity in patients with lower AFP levels. PMID- 15828958 TI - Study of a rat model for calcium oxalate crystal formation without severe renal damage in selected conditions. AB - BACKGROUND: Although nephrotoxic in high doses, ethylene glycol (EG) has been used with ammonium chloride (NH(4)Cl) or vitamin D(3) to study calcium oxalate stone formation in rat models. In the present study we used EG alone or with NH(4)Cl to study hyperoxaluria, crystaluria, and crystal attachment to renal epithelial cells in rats with minimal renal damage. METHODS: Six-week-old male Sprague-Dawley (SD) rats were given food and special drinking water. In experiment 1 the drinking water contained 1.0% NH(4)Cl plus four different concentrations of EG (0.8%, 0.4%, 0.2%, 0.1%). In experiment 2 the drinking water contained EG alone (0.8%, 0.4%, 0.2%, 0.1%). Urine was collected for 24 h before the rats were sacrificed. In experiment 1 the rats were sacrificed 5-13 days after starting the special water. In experiment 2 the rats were sacrificed 7-21 days after starting the special water. Bladder urine was also obtained. Blood and urine were tested for calcium, phosphorus, and creatinine. In addition, urine was tested for pH, oxalate and N-acetyl-beta-D glucosaminidase (NAG). Kidney sections were stained with hematoxylin-eosin, von Kossa and Pizzolato stain. Crystal morphology was determined using polarizing microscopy, and composition was determined using high-resolution X-ray powder diffraction. RESULTS: Experiment 1: Aggravation of renal function, an increase in urinary oxalate and NAG excretion, and crystals observed in the kidneys all correlated with EG concentration and length of drinking time. In bladder urine, calcium oxalate monohydrate (COM) crystals exceeded calcium oxalate dihydrate (COD) crystals. Experiment 2: Renal function remained unchanged. Oxalate excretion increased and NAG increased slightly. Crystals occurred only in the papillary tip region. Crystals in bladder urine were mostly COD. CONCLUSION: In the current rat model, calcium oxalate crystaluria could be induced without severe renal damage in selected cases. Either and/or both COM and COD might form and interact with kidney epithelium. We propose different experimental conditions to study the various phases of calcium oxalate stone formation in young male SD rats. PMID- 15828960 TI - Perinephric angiomyolipoma: a unique development pattern surrounding the kidney. AB - We report a case of a 31-year-old man with extrarenal angiomyolipoma of the perinephric space. He presented with asymptomatic macrohematuria. Computed tomography of the abdomen revealed a large perinephric mass which was separated from the right kidney and its unique growth appeared to have surrounded the kidney. Extrarenal angiomyolipomas of the perinephric fat are rare and they should be considered in the differential diagnosis of a retroperitoneal mass where asymptomatic macrohematuria was presented at the onset. PMID- 15828959 TI - Penile erectile responses to electric stimulation are enhanced by a new phosphodiesterase type-5 inhibitor. AB - AIM: This study was conducted to investigate the effect of DA-8159, a new phosphodiesterase type-5 (PDE5) inhibitor, on electrostimulation-induced penile erection in rats. METHODS: Intracavernous pressure (ICP) and arterial blood pressure (BP) were simultaneously recorded through electric pelvic-ganglion stimulation (2-10 Hz) after the oral administration of DA-8159 (3 or 10 mg/kg) in normal and streptozotocin-induced diabetic rats. Statistical analysis was performed on the maximal intracavernous pressure (ICP), detumescence time, maximal intracavernous pressure/blood pressure (ICP/BP) ratio, and the area under the curve (AUC) of the ICP/BP ratio. RESULTS: In normal and diabetic rats, electrical stimulation of the pelvic ganglion induced a frequency- and dose dependent increase in the intracavernous pressure. The ICP/BP ratio and the corresponding AUC values were also significantly and dose-dependently increased after DA-8159 administration. In addition, the detumescence time significantly increased after DA-8159 administration compared to that of the controls. CONCLUSIONS: These results show that the DA-8159 significantly increased the intracavernous pressure response and prolonged the decay period induced by electrical stimulation of the pelvic ganglion, and suggest that DA-8159 might be a potential therapeutic agent for the treatment of erectile dysfunction. PMID- 15828961 TI - Unilateral and segmental cystic disease of the kidney. AB - Unilateral and segmental cystic disease is a rare cystic disease of the kidney. Except for its unilaterality, it resembles autosomal dominant polycystic kidney disease on gross and histological examination. It also lacks genetic background and progressive deterioration of renal function. Only a small number of unilateral and segmental cystic disease of the kidney cases have been reported to date. The present case is a report of a 69-year-old man who had a large multicystic mass involving the lower and middle part of the right kidney with normal residual renal parenchyma at the upper pole. The left kidney was entirely normal. All the laboratory tests were within normal limits. Right nephrectomy was performed because of a strong suspicion of malignancy. PMID- 15828962 TI - Indirect computed tomography sign of renal artery injury: retrograde filling of the renal vein. AB - Computed tomography is a very useful diagnostic tool in children's trauma. In the present case report, retrograde filling of the renal vein during computerized tomographic examination of a patient with renal trauma is presented. This is an indirect sign of traumatic renal artery injury. This finding might assist in the early diagnosis of severe renovascular trauma. PMID- 15828963 TI - Successful management of lower pole moiety ureteropelvic junction obstruction in a partially duplicated collecting system using retrograde endoureteropyelotomy with the Holmium:YAG laser. AB - A patient with lower pole moiety ureteropelvic junction obstruction in a partially duplicated collecting system was managed successfully by retrograde endoureteropyelotomy using a Holmium:YAG laser. To our knowledge, we report the first case of this entity managed successfully by retrograde endoureteropyelotomy without a percutaneous approach. PMID- 15828964 TI - Transitional cell carcinoma of the bladder presenting after blunt abdominal trauma: a very rare occurrence in childhood. AB - Tumors of the urinary bladder are rare in children and nearly all of them are of mesodermal in origin. Herein we report a case of transitional cell carcinoma of the urinary bladder in a 13-year-old boy who presented after blunt abdominal trauma. It should be remembered that hematuria could develop after blunt abdominal trauma in children, for reasons other than the trauma itself (e.g. tumors of the urinary tract). PMID- 15828965 TI - Adenosquamous carcinoma of the prostate. AB - Adenosquamous carcinoma of the prostate is an unusual histological variant of prostate cancer. The histogenesis of this tumor remains uncertain. The stimulus for the development of the squamous metaplastic cells had been thought to be related to hormone and/or radiation therapy. This report presents a case of adenosquamous carcinoma of the prostate with abscence of previous hormone or radiation therapy. The case showed negative prostate-specific antigen and high molecular weight cytokeratin staining of the adenocarcinoma component, and negative prostate-specific antigen and positive high molecular weight cytokeratin staining of the squamous cell carcinoma component. The adenocarcinoma component stained intraluminally with periodic acid schiff. The staining features and the distinct localizations of the components with intermingling, but no transition, are against the collision-type tumor theory and support the theory that the adenocarcinoma and squamous components arise de novo from pluripotent stem cells. The patient had a rapid downhill clinical course and died 3 weeks after the diagnosis was made. PMID- 15828966 TI - Recto-peritoneal fistula following transperineal prostate biopsy. AB - A recto-peritoneal fistula is an extremely rare complication after prostate biopsy. We report herein on a peritonitis arising from a recto-peritoneal fistula 5 days after undergoing prostate biopsy. To our knowledge, this is the first case of recto-peritoneal fistula following transperineal needle biopsy of the prostate in the published literature. PMID- 15828967 TI - Use of atresic vagina as a urethra to repair iatrogenic incontinence, secondary to a complete longitudinal urethral incision. AB - Vaginal atresia is a rare congenital anomaly. Patients with this pathology commonly present with amenorrhea, or an inability to have sexual intercourse. A dorsal urethral wall incision was performed in a woman who complained of difficulty with coitus. Her gynecologist intended to expand the vagina, thinking the diagnosis was a vaginal septum. We report a repair technique for total urinary incontinence following a longitudinal complete urethral incision, which also involved the bladder neck. The vaginal reconstruction consisted of an ileal segment, whereas the urinary incontinence was corrected using a rectus fascial sling. To our knowledge, this is the first case of this kind reported in the literature. PMID- 15828968 TI - Primary carcinoid tumor of the testis with teratoma metastatic to the para-aortic lymph node. AB - A primary testicular carcinoid tumor with teratoma metastasized to the para aortic lymph node. After inguinal orchiectomy, serum and urinary levels of 5 hydroxyindoleacetic acid (5-HIAA) were found to be elevated. Chemotherapy consisting of cisplatin, etoposide and bleomycin was not effective on the metastatic lesions. Retroperitoneal lymphadenectomy normalized the levels of 5 HIAA. The patient is alive without recurrence 25 months after the retroperitoneal lymphadenectomy. PMID- 15828969 TI - Unusual presentation of necrotizing fasciitis in a patient who had achieved long term remission after irradiation for testicular cancer. AB - We report a case of a 60-year-old man with necrotizing fasciitis complicated by streptococcal toxic shock syndrome. The patient had received high-dose chemotherapy and radiotherapy to the pelvis for relapsed seminoma 7 years previously. He had been in long-term remission. He was admitted to the Tsukuba University Hospital, Tsukuba-City, Ibaraki, Japan, with complaints of fever and localized erythema over the foreskin. The patient suffered from septic shock and multiple organ failure. Despite intensive care, he died 18 h after admission. Streptococcus pyogenes was isolated from both the wound and blood culture. To our knowledge, this is the first description of necrotizing fasciitis primarily affecting the penile skin. PMID- 15828970 TI - Toddlers' representations of space: the role of viewer perspective. AB - In the present study, we examined how people code object location when an object is hidden in an enclosed space. On object location tasks involving disorientation, viewers must code the location of an object in relation to the spatial environment because they cannot directly track their changing relation to the object. Recently, we showed that viewers also code their perspective relative to the entire space (inside vs. outside) in such tasks. To determine whether viewers code perspective whenever they must locate an object, we examined young children's performance on a task in which coding perspective would be difficult. Our task involved both disorientation and translation (i.e., movement from inside to outside the space or vice versa). When translation preceded disorientation, performance was comparable to performance on tasks with no translation. However, when disorientation preceded translation, performance was at chance. The implications of these findings for location coding are discussed. PMID- 15828971 TI - Ambiguous pronoun resolution: contrasting the first-mention and subject preference accounts. AB - A visual-world eye-tracking experiment investigated the influence of order of mention and grammatical role on resolution of ambiguous pronouns in Finnish. According to the first-mention account, general cognitive structure-building processes make the first-mentioned noun phrase the preferred antecedent of an ambiguous pronoun. According to the subject-preference account, the preferred antecedent is the grammatical subject of the preceding clause or sentence. Participants listened to sentences in either subject-verb-object or object-verb subject order; each was followed by a sentence containing an ambiguous pronoun that referred to either the subject or the object. Participants' eye movements were monitored while they looked at pictures representing the two possible antecedents of each pronoun. Analyses of the fixations on the pictures showed that listeners used both order-of-mention and grammatical-role information to resolve ambiguous pronouns. PMID- 15828972 TI - The beneficial effect of concurrent task-irrelevant mental activity on temporal attention. AB - It is believed that the human cognitive system is fundamentally limited in deploying attention over time. This limitation is reflected in the attentional blink, the impaired ability to identify the second of two visual targets presented in close succession. We report the paradoxical finding that the attentional blink is significantly ameliorated when observers are concurrently engaged in distracting mental activity, such as free-associating on a task irrelevant theme or listening to music. This finding raises questions about the fundamental nature of the attentional blink, and suggests that the temporal dynamics of attention are determined by task circumstances that induce either a more or a less distributed state of mind. PMID- 15828973 TI - Object onset and parvocellular guidance of attentional allocation. AB - The parvocellular visual pathway in the primate brain is known to be involved with the processing of color. However, a subject of debate is whether an abrupt change in color, conveyed via this pathway, is capable of automatically attracting attention. It has been shown that the appearance of new objects defined solely by color is indeed capable of modulating attention. However, given evidence suggesting that the visual system is particularly sensitive to new onsets, it is unclear to what extent such results reflect effects of color change per se, rather than effects of object onset. We assessed attentional capture by color change that occurred as a result of either new objects appearing or already present "old" objects changing color. Results showed that although new object onsets accrued attention, changing the color of old objects did not. We conclude that abrupt color change per se is not sufficient to capture attention. PMID- 15828974 TI - Do new objects capture attention? AB - The visual system relies on several heuristics to direct attention to important locations and objects. One of these mechanisms directs attention to sudden changes in the environment. Although a substantial body of research suggests that this capture of attention occurs only for the abrupt appearance of a new perceptual object, more recent evidence shows that some luminance-based transients (e.g., motion and looming) and some types of brightness change also capture attention. These findings show that new objects are not necessary for attention capture. The present study tested whether they are even sufficient. That is, does a new object attract attention because the visual system is sensitive to new objects or because it is sensitive to the transients that new objects create? In two experiments using a visual search task, new objects did not capture attention unless they created a strong local luminance transient. PMID- 15828975 TI - Microgenesis and ontogenesis of perceptual organization: evidence from global and local processing of hierarchical patterns. AB - In two experiments, visual search and speeded classification were used to study perception of hierarchical patterns among participants aged 5 to 23 years. Perception of global configurations of few-element patterns and local elements of many-element patterns showed large age-related improvements. Only minor age related changes were observed in perception of global configurations of many element patterns and local elements of few-element patterns. These results are consistent with prior microgenetic analyses using hierarchical patterns. On the one hand, the rapid and effortless grouping of many small elements and the individuation of few large elements both mature by age 5. In contrast, the time consuming and effortful grouping of few large elements and the individuation of many small elements improve substantially with age, primarily between ages 5 and 10. These findings support the view that perceptual organization involves multiple processes that vary in time course, attentional demands, and developmental trajectories. PMID- 15828976 TI - Not quite as grown-up as we like to think: parallels between cognition in childhood and adulthood. AB - Greater continuity in cognition between children and adults may exist than is usually appreciated. It was thought that after 3 to 4 years of age, the problem in switching on the dimensional-change card-sort task disappears. We show here, however, that if speed is used as the dependent measure, the effect of the first dimension is evident even in adults. Adults, like preschoolers, show difficulty in switching from a block of sorting by color or shape to a block of sorting by the other dimension. Notably, performance throughout the session was affected by the first dimension by which stimuli were sorted. We hypothesize that perhaps adults never fully outgrow any of the cognitive and perceptual biases of infancy and early childhood. Other examples of such biases that appear to still be present in adults are discussed. Conversely, the assumption that the optimal dependent measure for adults is the most sensitive measure for children is questioned. PMID- 15828978 TI - Temporally nonadjacent nonlinguistic sounds affect speech categorization. AB - Speech perception is an ecologically important example of the highly context dependent nature of perception; adjacent speech, and even nonspeech, sounds influence how listeners categorize speech. Some theories emphasize linguistic or articulation-based processes in speech-elicited context effects and peripheral (cochlear) auditory perceptual interactions in non-speech-elicited context effects. The present studies challenge this division. Results of three experiments indicate that acoustic histories composed of sine-wave tones drawn from spectral distributions with different mean frequencies robustly affect speech categorization. These context effects were observed even when the acoustic context temporally adjacent to the speech stimulus was held constant and when more than a second of silence or multiple intervening sounds separated the nonlinguistic acoustic context and speech targets. These experiments indicate that speech categorization is sensitive to statistical distributions of spectral information, even if the distributions are composed of nonlinguistic elements. Acoustic context need be neither linguistic nor local to influence speech perception. PMID- 15828977 TI - Mommy and me: familiar names help launch babies into speech-stream segmentation. AB - How do infants find the words in the tangle of speech that confronts them? The present study shows that by as early as 6 months of age, infants can already exploit highly familiar words-including, but not limited to, their own names-to segment and recognize adjoining, previously unfamiliar words from fluent speech. The head-turn preference procedure was used to familiarize babies with short passages in which a novel word was preceded by a familiar or a novel name. At test, babies recognized the word that followed the familiar name, but not the word that followed the novel name. This is the youngest age at which infants have been shown capable of segmenting fluent speech. Young infants have a powerful aid available to them for cracking the speech code. Their emerging familiarity with particular words, such as their own and other people's names, can provide initial anchors in the speech stream. PMID- 15828979 TI - Touch noise increases vibrotactile sensitivity in old and young. AB - Stochastic resonance (SR) occurs when the detection of a subthreshold signal is aided by the presence of random energy fluctuations in the signal modality, commonly called noise. SR is counterintuitive because such noise usually worsens performance. Nonetheless, SR has been demonstrated both theoretically and experimentally in human sensory systems. Using a psychophysically sophisticated paradigm, we show that SR aids the detection of vibrating touch stimuli presented to the foot soles of both healthy elderly people with elevated vibrotactile thresholds and healthy young people with normal vibrotactile thresholds. The results also suggest that it is possible to know a priori the amount of noise needed for optimal SR effects given the degree to which the signal is subthreshold. Thus, SR may be practical as a rehabilitative aid for individuals with elevated sensory thresholds. PMID- 15828980 TI - Color naming, lens aging, and grue: what the optics of the aging eye can teach us about color language. AB - Many languages without separate terms for green and blue are or were spoken in locations receiving above-average exposure to ultraviolet-B (UV-B) radiation. It has been proposed that this correlation is caused by premature lens aging. This conclusion was supported by an experiment in which younger observers used the term "blue" less often when they described simulated paint chips filtered through the equivalent of an older observer's lens-removing much short-wavelength light than when they described the unfiltered versions of the same paint chips. Some stimuli that were called "blue" without simulated aging were called "green" when filtered. However, in the experiment reported here, we found that the proportion of "blue" color-name responses did not differ between younger subjects and older observers with known ocular media optical densities. Color naming for stimuli that were nominally green, blue-green, or blue was virtually identical for older and younger observers who viewed the same (unfiltered) stimuli. Our results are inconsistent with the lens-brunescence hypothesis. PMID- 15828981 TI - Low self-esteem is related to aggression, antisocial behavior, and delinquency. AB - The present research explored the controversial link between global self-esteem and externalizing problems such as aggression, antisocial behavior, and delinquency. In three studies, we found a robust relation between low self-esteem and externalizing problems. This relation held for measures of self-esteem and externalizing problems based on self-report, teachers' ratings, and parents' ratings, and for participants from different nationalities (United States and New Zealand) and age groups (adolescents and college students). Moreover, this relation held both cross-sectionally and longitudinally and after controlling for potential confounding variables such as supportive parenting, parent-child and peer relationships, achievement-test scores, socioeconomic status, and IQ. In addition, the effect of self-esteem on aggression was independent of narcissism, an important finding given recent claims that individuals who are narcissistic, not low in self-esteem, are aggressive. Discussion focuses on clarifying the relations among self-esteem, narcissism, and externalizing problems. PMID- 15828982 TI - Forgetting of trauma cues in adults reporting continuous or recovered memories of childhood sexual abuse. AB - According to betrayal trauma theory, adult survivors of childhood sexual abuse (CSA) who were molested by their caretakers (e.g., a father) are especially likely to dissociate ("repress") their memories of abuse. Testing college students, some reporting CSA, DePrince and Freyd (2004) found that those scoring high on a dissociation questionnaire exhibited memory deficits for trauma words when they viewed these words under divided-attention conditions. Replicating DePrince and Freyd's procedure, we tested for memory deficits for trauma words relative to neutral words in adults reporting either continuous or recovered memories of CSA versus adults denying a history of CSA. A memory deficit for trauma words under divided attention was expected in the recovered-memory group. Results were inconsistent with this prediction, as all three groups exhibited better recall of trauma words than neutral words, irrespective of encoding conditions. PMID- 15828983 TI - New approaches to using census data to test the critical-period hypothesis for second-language acquisition. PMID- 15828984 TI - Brute force or gentle persuasion? PMID- 15828985 TI - The use of steroids in children with septicemia: review of the literature and assessment of current practice in PICUs in the UK. PMID- 15828986 TI - Mask fear in children presenting for anesthesia: aversion, phobia, or both? AB - BACKGROUND: Anesthesia induction in children is commonly accomplished by introducing volatile agents by mask. Occasionally a child describes an excessive fear of the anesthesia facemask. Little is known of the cause of the fear or of the quality or magnitude of the feelings the child is experiencing. The purpose of this study was to allow children who have established mask fear as demonstrated by volunteering the presence of fear and requesting no mask be placed on the face during the induction of anesthesia and their parents to describe and compare the distress from the mask to the alternative intravenous anesthesia induction. METHODS: Eight children describing mask fear on the preanesthetic examination were studied. An Anesthesia Mask Fear questionnaire developed by the investigators was answered by the children and their parents. RESULTS: Six children and their parents completed the study. The age at presentation of mask fear ranged from 1.4 to 14 years. There were one to 16 anesthetic exposures prior to reporting mask fear. One child described an aversion to the odor of the mask. Another boy developed mask fear after a single anesthetic exposure. He was subsequently diagnosed with a generalized anxiety disorder. Four female children developed mask fear after repeated anesthetic exposures. These children rated mask fear with the greatest discomfort possible while venous cannulation was scored at half or less that of the mask discomfort. CONCLUSIONS: Care must be taken when developing a plan for anesthesia induction in children requiring multiple procedures. Children may develop an aversion to the odor or feel of the mask, or have a true phobia (irrational fear) of the mask. Those children with a phobia might also have other underlying anxieties. PMID- 15828987 TI - Anatomical considerations of the pediatric ilioinguinal/iliohypogastric nerve block. AB - BACKGROUND: The ilioinguinal/iliohypogastric nerve block is safe, effective and easy to perform in order to provide analgesia for a variety of inguinal surgical procedures in pediatric patients. A relatively high failure rate of 10-25% has been reported, even in experienced hands. The aim of this study was to determine the exact anatomical position of the ilioinguinal and iliohypogastric nerves in relation to an easily identifiable constant bony landmark, the anterior superior iliac spine (ASIS) in neonates and infants. The current ilioinguinal/iliohypogastric nerve block techniques were also evaluated from an anatomical perspective. METHOD: Dissections were performed on a sample of 25 infant and neonatal cadavers (mean weight = 2.2 kg; mean height = 45.6 cm). The distance from the ASIS to both the ilioinguinal and iliohypogastric nerves, on a line connecting the ASIS to the umbilicus was carefully measured using a digital caliper. Three techniques, commonly used in clinical practice, were simulated on the anatomical specimens. RESULT: The left and right ilioinguinal nerves were closer to the ASIS than previously described, i.e. 1.9 +/- 0.9 mm (mean +/- sd) and 2.0 +/- 0.7 mm, respectively. The mean distance from the left and right iliohypogastric nerves to the ASIS are 3.3 +/- 0.8 mm and 3.9 +/- 1.0 mm, respectively. CONCLUSIONS: We suggest that the high failure rate of the ilioinguinal/iliohypogastric nerve block in this age group could be due to lack of specific spatial knowledge of the anatomy of these nerves in infants and neonates. This cadaver-based study suggests an insertion point closer to the ASIS, approximately 2.5 mm (range: 1.0-4.9) from the ASIS on a line drawn between the ipsilateral ASIS and the umbilicus. PMID- 15828988 TI - The correlation of the Narcotrend Index and classical electroencephalographic parameters with endtidal desflurane concentrations and hemodynamic parameters in different age groups. AB - BACKGROUND: The Narcotrend Index (NI) for assessment of depth of anesthesia by analysis of the electroencephalogram (EEG), is potentially a pharmacodynamic measure of the effects of desflurane on the brain. METHODS: In this prospective study of 30 pediatric and adult patients (group 1: 3-6 years, n = 10; group 2: > 6 < 12 years; group 3: 12-40 years), undergoing ophthalmological surgery, we investigated the pharmacodynamic relationship between nonsteady state endtidal desflurane concentrations (eT(Des)), NI, classical EEG parameters (cEEG), heart rate (HR) and mean arterial pressure (MAP). The performance of the Narcotrend for differentiation between consciousness and unconsciousness was evaluated using prediction probability (P(K)). RESULTS: Spearman correlation analysis showed significant negative correlations (P < 0.001) between eT(Des) and NI (group 1: r = -0.93, group 2: r = -0.86, group 3: r = -0.66). Correlations between eT(Des) and MAP or HR were either only weak negative (r < -0.5) or not significant. Desflurane EC(50) (eT(Des) with half maximal effect on NI) was 7.18% for group 1, 7.34% for group 2, and 4.15% for group 3 (P < 0.001 Vs groups 1 and 2). Overall awake NI values (96.7 +/- 1.4) were significantly higher (P < 0.001) than at the moment of loss of consciousness (58.3 +/- 17.5), with no overlap (P(K) 1.0), whereas P(K) values for cEEG, MAP and HR were all <0.85. CONCLUSIONS: The pharmacodynamic relationship between eT(Des) and NI is age dependent with a significantly higher EC(50) in children than in adolescents and adults. The NI appears to be superior to cEEG, MAP and HR in differentiating consciousness from unconsciousness. PMID- 15828989 TI - Ventilation with the laryngeal tube in pediatric patients undergoing elective ambulatory surgery. AB - BACKGROUND: The laryngeal tube is a new supraglottic alternative for securing the airway. Few data on the use of this airway device in pediatric patients are so far available. Experiences of the study are reported. METHODS: Use of the laryngeal tube for ventilation during intermittent positive-pressure ventilation was studied in an open, noncomparative observational study in 80 pediatric patients (age: 2-12 years) undergoing elective ambulatory surgery. Insertion success, time to achieve a patent airway, and occurrence of adverse events were documented. RESULTS: Mean (sd) patient age was 5.8 (2.3) years. The laryngeal tube was placed at the first attempt in 90% of patients, a second attempt was necessary in 6%. Ventilation was not possible after two attempts in three patients. Mean (sd) tidal volume was 10.1 (2.0) ml.kg(-1) with a mean (sd) peak airway pressure of 15.6 (2.1) cmH(2)O. Chin lift was applied to improve ventilation in 15 patients (18.8%). Laryngospasm occurred in one patient, but ventilation was possible after adapting depth of anesthesia. Mean (sd) time for airway maintenance was 11.2 (3.7) s. No gastric insufflation of air occurred, no traces of blood could be detected after removal of the device. One patient complained of mild problems (swallowing, VAS 1). Mean (sd) anesthesia time was 44 (9) min. CONCLUSIONS: In the age group studied, the laryngeal tube provides a rapid, patent airway in the majority of patients with a low complication rate. PMID- 15828990 TI - Clinical assessment of the laryngeal tube in pediatric anesthesia. AB - BACKGROUND: The aim of this study was to evaluate a new device for airway management in children: the laryngeal tube (LT). METHODS: The LT is available in sizes S0-S3 for pediatric anesthesia. This prospective open study included 70 children ASA 1. The local Ethics Committee approval and parental consent were collected. The primary criterion was the success rate for insertion and ventilation. Secondary criteria were additional maneuvers and incidents elicited from LT use. RESULTS: Seventy children were included: S0 = 5, S1 = 8, S2 = 36, and S3 = 21. Insertion was successful: at the first attempt in 78.6%, second in 17.1%, and third or more in 4.3%. In 12% of cases it was not possible to successfully insert the LT and proceed to adequate ventilation. Failures were explained by: inability to obtain satisfying ventilation (n = 4), hypoxemia (n = 1), gastric insufflation (n = 6), cough (n = 1), and laryngospasm or stridor (n = 2), some with the same child. Minimal additional maneuvers for adequate ventilation were necessary in 35% of cases (all groups), but <20% when considering only sizes 2 and 3. Moreover, after five cases, the anesthesiologists became more proficient at inserting the LT (respectively 73.3% failure before five cases vs 13% afterwards). Gastric insufflation occurred in eight cases (11.4%). Controlled ventilation was used in 30 children and peak inspiratory pressure was 19.2 +/- 4 cmH(2)O. CONCLUSIONS: The LT is not recommended for children <10 kg. Over 10 kg, it provides a clear airway in most children, with a low rate of minimal additional maneuvers for sizes 2 and 3. The failure rate also decreases with the operator's training. PMID- 15828991 TI - Combined propofol and remifentanil intravenous anesthesia for pediatric patients undergoing magnetic resonance imaging. AB - BACKGROUND: A prospective observational case series of children receiving light general anesthesia for magnetic resonance imaging (MRI) was performed. Our purpose was to examine the merit of anesthesia and recovery/discharge times of combined remifentanil and propofol total intravenous anesthesia (TIVA) in spontaneously breathing children. METHODS: After IRB approval and informed consent, 56 patients receiving Remi/Propofol TIVA (Remifentanil 10 microg.ml(-1) Propofol 10 mg.ml(-1)) were observed. Blood pressure, respiratory rate, endtidal CO(2) (P(E)CO(2)), oxygen saturation and temperature were recorded at the start and finish of anesthesia. In addition, induction and recovery times were noted. Recovery time was from scan completion until discharge from the initial recovery area. Discharge time was from scan completion to discharge home. RESULTS: Fifty six patients received Remi/Propofol TIVA. The mean Remi/Propofol recovery and discharge times were 8.9 and 28.2 min, respectively. There was a statistically significant decrease in respiratory rate and increase in CO(2) from the start to the end of the procedure. During the scan, seven patients moved. One patient experienced postprocedure nausea and or vomiting. CONCLUSIONS: The combination of remifentanil and propofol for TIVA may be an effective method of light general anesthesia in pediatric patients undergoing MRI. PMID- 15828992 TI - Dimensions of the neonatal cricothyroid membrane - how feasible is a surgical cricothyroidotomy? AB - BACKGROUND: Airway management of the neonate remains a cornerstone in neonatal resuscitation which in most cases involves tracheal intubation. However, difficult intubations do occur. Cricothyroidotomy is recognized as an entry point below the vocal cords. This procedure becomes increasingly difficult in young children and is not recommended in children under the age of 5 years. Little is known about the anatomy of the neonatal airway, especially the size of the cricothyroid membrane. The aim of the study was to determine the dimensions of the cricothyroid membrane in neonates. METHODS: Twenty-seven neonatal cadavers (mean height of 44.89 cm and a mean weight of 2.05 kg) were carefully dissected and the dimensions of the cricothyroid membrane recorded with a digital caliper (accuracy 0.01 mm) by two independent observers. RESULTS: The cricothyroid membrane has a mean height of 2.61 mm (sd: 0.71) and width of 3.03 mm (sd: 0.63). CONCLUSIONS: Results of this study indicate that the dimensions of the cricothyroid membrane are too small for passing a tracheal tube as the dimensions of the tube exceeds that of the cricothyroid membrane. This could fracture the cartilages of the larynx. The performance of a surgical cricothyroidotomy with passing of a tracheal tube is therefore strongly discouraged in neonatal patients. PMID- 15828993 TI - The use of physical restraints on paediatric intensive care units. AB - BACKGROUND: Physical restraints are used in critical care units in an attempt to reduce the risk of treatment interference. Their use remains controversial and there are concerns regarding the effectiveness and safety of restraint techniques. There are few data available on the prevalence of physical restraint use in Paediatric Intensive Care Units (PICU) in the UK and we have therefore conducted a cross-sectional survey to define current clinical practice. METHODS: A postal questionnaire was sent to all UK PICU with questions on the use of physical restraint techniques, including the use of splints across the joints of limbs and of securing limbs to the bed. Consent issues were also addressed. RESULTS: Of those units responding 68% reported that physical restraint techniques were used within those units. Ten units (36% of those responding) reported the use of manual holding of patients, 16 units (57% of those responding) reported the use of splints across the joints of limbs, three units reported the use of swaddling as a restraint technique while one unit secured limbs of patients to the bed. None of the units obtained written consent prior to the use of physical restraints and 53% obtained verbal consent. CONCLUSIONS: Physical restraint is a commonly used technique on PICU in the UK. There is considerable variation in clinical practice and current clinical guidelines which are available do not deal specifically with children. Prospective randomized trials would be necessary to fully investigate the role of physical restraints amongst critically ill children. PMID- 15828994 TI - Neuropathic pain following multilevel surgery in children with cerebral palsy: a case series and review. AB - Six children with cerebral palsy are presented who developed neuropathic pain following multilevel orthopedic surgery. This significant complication is previously unreported. The diagnosis and treatment options are reviewed. Treatment should be kept as simple and noninvasive as possible, and aim to enable physiotherapy to continue. Early recognition and interdisciplinary treatment is important to prevent a downward spiral of increasing pain and decreased function. A good outcome in respect of improved pain and functioning was achieved in five of these six children. It is our opinion that this complication should form part of informed consent for multilevel surgery and that anesthetists should be aware of this complication when managing postoperative pain control. PMID- 15828995 TI - Induction of anesthesia in a combative child; management and issues. AB - A developmentally delayed, 13-year old autistic boy required management of multifocal cerebral and pulmonary tumors, involving several anesthetics over a 4 month period. At each anesthetic he refused premedication, displayed increasing anxiety and became more combative. With parental guidance and involvement, a variety of anesthetists tried a range of techniques to achieve induction, each ultimately resorting to the use of physical restraint. Principles essential to the care of such a child include early recognition, parental support, multi disciplinary planning of procedures requiring general anesthesia, continuity of anesthesia care, and clear guidelines about the perioperative management of uncooperative children, including the ethical use of restraint. PMID- 15828996 TI - Anesthesia in a child with homozygous porphobilinogen deaminase deficiency: a severe form of acute intermittent porphyria. AB - We report a case history of the anesthetic management of a child with a severe form of acute intermittent porphyria (AIP). AIP is an autosomal dominant condition with incomplete penetrance, caused by deficiency of porphobilinogen deaminase, an enzyme found in the synthetic pathway for heme. Anesthesia and surgery may present many precipitants for a potentially fatal acute porphyric attack. These include fasting, dehydration, stress, infection and drugs. Here, we describe the safe use of sevoflurane in the maintenance of anesthesia. Its relative insolubility and low metabolism suggest that sevoflurane may be a reasonable agent for anesthesia in the porphyric patient. PMID- 15828997 TI - Ultrasound assessment of caudal catheter position in infants. AB - The positioning of caudally inserted epidural catheters is crucial to their effectiveness. However, level assessment can be difficult and time consuming. We report the use of ultrasound to assess the catheter position in three patients aged between 1 and 10 months. The advantages and disadvantages of this technique are discussed in relation to other methods of assessing caudal catheter placement. PMID- 15828998 TI - Anesthetic management in Friedreich's ataxia. PMID- 15828999 TI - Delayed awakening from general anesthesia in a hypovolemic infant. PMID- 15829000 TI - Mediastinal mass syndrome. PMID- 15829002 TI - Freeman-Sheldon syndrome. PMID- 15829003 TI - Inadvertent median artery cannulation. PMID- 15829004 TI - Ultrasound image attributes of human ovarian dominant follicles during natural and oral contraceptive cycles. AB - BACKGROUND: Computer-assisted analyses were used to examine ultrasound image attributes of human dominant ovarian follicles that developed during natural and oral contraceptive (OC) cycles. We hypothesized that image attributes of natural cycle follicles would quantitatively differ from those in OC cycles and that OC cycle follicles would possess image attributes indicative of atresia. METHODS: Dominant ovarian follicles of 18 clinically normal women were compared using transvaginal ultrasonography for the 7 days before ovulation during a natural cycle (n = 9) or the 7 days before peak estradiol in women using OC (n = 11). Follicles were analyzed using region and line techniques designed to compare the image attributes numerical pixel value (NPV), pixel heterogeneity (PH) and area under the curve (AUC). RESULTS: NPV was higher in OC cycle follicles with region analysis and tended to be higher with line analysis (p = 0.005 and p = 0.06, respectively). No differences were observed in two other image attributes (AUC and PH), measured with either technique, between natural and OC cycle follicles. CONCLUSION: The increased NPV value of OC cycle follicles and lack of differences in PH and AUC values between natural cycle and OC cycle follicles did not support the hypothesis that OC cycle follicles would show ultrasonographically detectable signs of atresia. Image attributes observed in OC cycle follicles were not clearly indicative of atresia nor were they large enough to preclude preovulatory physiologic status in OC cycle follicles. PMID- 15829005 TI - Validation of multi-stage telephone-based identification of cognitive impairment and dementia. AB - BACKGROUND: Many types of research on dementia and cognitive impairment require large sample sizes. Detailed in-person assessment using batteries of neuropyschologic testing is expensive. This study evaluates whether a brief telephone cognitive assessment strategy can reliably classify cognitive status when compared to an in-person "gold-standard" clinical assessment. METHODS: The gold standard assessment of cognitive status was conducted at the University of Southern California Alzheimer Disease Research Center (USC ADRC). It involved an examination of patients with a memory complaint by a neurologist or psychiatrist specializing in cognitive disorders and administration of a battery of neuropsychologic tests. The method being evaluated was a multi-staged assessment using the Telephone Interview of Cognitive Status-modified (TICSm) with patients and the Telephone Dementia Questionnaire (TDQ) with a proxy. Elderly male and female patients who had received the gold standard in-person assessment were asked to also undergo the telephone assessment. The unweighted kappa statistic was calculated to compare the gold standard and the multistage telephone assessment methods. Sensitivity for classification with dementia and specificity for classification as normal were also calculated. RESULTS: Of 50 patients who underwent the gold standard assessment and were referred for telephone assessment, 38 (76%) completed the TICS. The mean age was 78.1 years and 26 (68%) were female. When comparing the gold standard assessment and the telephone method for classifying subjects as having dementia or no dementia, the sensitivity of the telephone method was 0.83 (95% confidence interval 0.36, 1.00), the specificity was 1.00 (95% confidence interval 0.89,1.00). Kappa was 0.89 (95% confidence interval 0.69, 1.000). Considering a gold-standard assessment of age associated memory impairment as cognitive impairment, the sensitivity of the telephone approach is 0.38 (95% confidence interval 0.09, 0.76) specificity 0.96 (CI 0.45, 0.89) and kappa 0.61 (CI 0.37, 0.85). CONCLUSION: Use of a telephone interview to identify people with dementia or cognitive impairment is a promising and relatively inexpensive strategy for identifying potential participants in intervention and clinical research studies and for classifying subjects in epidemiologic studies. PMID- 15829006 TI - Difficulties in assessing cytomegalovirus-associated gastric perforation in an HIV-infected patient. AB - BACKGROUND: Active Cytomegalovirus (CMV) infection is a common complication in advanced symptomatic Human Immunodeficiency Virus (HIV) infection. CMV-induced intestinal perforations are hard to diagnose and may be observed throughout the gastrointestinal tract. Isolated stomach perforation is exceptional. CASE PRESENTATION: A 47-year-old man was admitted to our intensive care unit with multiorgan failure. Gastrointestinal endoscopic examination showed erythematous gastritis but normal duodenum and colon. CMV blood culture was positive. Histologic examination of a gastric biopsy showed inflammatory infiltrate and immunostaining typical intranuclear CMV inclusion bodies. Concomitant abdominal CT scan disclosed large peripancreatic hypodensities without pneumoperitoneum. The patient died despite supportive therapies and ganciclovir infusion. Postmortem examination showed a 4-cm gastric perforation adhering to the transverse colon and liver, with a thick necrotic inflammatory coating around the pancreas. The whole GI tract, except the stomach, was normal. As other causes, especially Helicobacter pylori infection could be ruled out, a causal relationship between CMV and gastric disease was assumed. CONCLUSION: CMV may be responsible for gastric perforations, with difficulties in assessing the diagnosis. Early diagnosis based on cautious endoscopy and histopathologic examination is needed to make a favorable outcome possible. PMID- 15829007 TI - ProGenExpress: visualization of quantitative data on prokaryotic genomes. AB - BACKGROUND: The integration of genomic information with quantitative experimental data is a key component of systems biology. An increasing number of microbial genomes are being sequenced, leading to an increasing amount of data from post genomics technologies. The genomes of prokaryotes contain many structures of interest, such as operons, pathogenicity islands and prophage sequences, whose behaviour is of interest during infection and disease. There is a need for simple and novel tools to display and analyse data from these integrated datasets, and we have developed ProGenExpress as a tool for visualising arbitrarily complex numerical data in the context of prokaryotic genomes. RESULTS: Here we describe ProGenExpress, an R package that allows researchers to easily and quickly visualize quantitative measurements, such as those produced by microarray experiments, in the context of the genome organization of sequenced prokaryotes. Data from microarrays, proteomics or other whole-genome technologies can be accurately displayed on the genome. ProGenExpress can also search for novel regions of interest that consist of groups of adjacent genes that show similar patterns across the experimental data set. We demonstrate ProGenExpress with microarray data from a time-course experiment involving Salmonella typhimurium. CONCLUSION: ProGenExpress can be used to visualize quantitative data from complex experiments in the context of the genome of sequenced prokaryotes, and to find novel regions of interest. PMID- 15829008 TI - Prevalence and risk factors for Plasmodium falciparum malaria in pregnant women of eastern Sudan. AB - BACKGROUND: Pregnant women are more susceptible to malaria, which is associated with serious adverse effects on pregnancy. The presentation of malaria during pregnancy varies according to the level of transmission in the area. Our study aimed to demonstrate the prevalence and risk factors for malaria (age, parity and gestational age) among pregnant women of eastern Sudan, which is characterized by unstable malaria transmission. METHODS: The prevalence and possible risk factors for Plasmodium falciparum malaria were investigated in 744 pregnant Sudanese women attending the antenatal clinic of New Haifa Teaching Hospital, eastern Sudan, during October 2003-April 2004. RESULTS: A total 102 (13.7%) had P. falciparum malaria, 18(17.6%) of these were severe cases (jaundice and severe anaemia). Univariate and multivariate analysis showed that, age and parity were not associated with malaria. Women who attended the antenatal clinic in the third trimester were at highest risk for malaria (OR = 1.58, 95% CI = 1.02-2.4; P < 0.05). Women with malaria had significantly lower mean haemoglobin (9.4 g/dl, 95% CI 9.1-9.7 versus 10.7, CI 10.6-10.8, P < 0.05). A significantly lower haemoglobin was observed in those with severe falciparum malaria compared to non severe form (8.3 g/dl, 95% CI 7.6-9.1 versus 9.4, 95% CI 9.1-9.7, P = or < 0.05). CONCLUSION: The results suggest that P. falciparum malaria is common in pregnant women attending antenatal care and that anaemia is an important complication. Preventive measures (chemoprophylaxis and insecticide-treated bednets) may be beneficial in this area for all women irrespective of age or parity. PMID- 15829009 TI - The accuracy of the report of hepatic steatosis on ultrasonography in patients infected with hepatitis C in a clinical setting: a retrospective observational study. AB - BACKGROUND: Steatosis is occasionally reported during screening ultrasonography in patients with hepatitis C virus (HCV). We conducted a retrospective observational study to assess the factors associated with steatosis on ultrasonography and the relationship between steatosis on ultrasound versus biopsy in patients infected with HCV in a clinical setting. Our hypothesis was ultrasonography would perform poorly for the detection of steatosis outside of the context of a controlled study, primarily due to false-positive results caused by hepatic fibrosis and inflammation. METHODS: A retrospective review of ultrasound reports was conducted on patients infected with HCV in a tertiary care gastroenterology clinic. Reports were reviewed for the specific documentation of the presence of steatosis. Baseline clinical and histologic parameters were recorded, and compared for patients with vs. without steatosis. Multiple logistic regression analysis was performed on these baseline variables. Liver biopsies were reviewed by two pathologists, and graded for steatosis. Steatosis on biopsy was compared to steatosis on ultrasound report, and the performance characteristics of ultrasonography were calculated, using biopsy as the gold standard. RESULTS: Ultrasound reports were available on 164 patients. Patients with steatosis on ultrasound had a higher incidence of the following parameters compared to patients without steatosis: diabetes (12/49 [24%] vs. 7/115 [6%], p < 0.001), fibrosis stage > 2 (15/48 [31%] vs. 16/110 [15%], p = 0.02), histologic grade > 2 (19/48 [40%] vs. 17/103 [17%], p = 0.002), and ALT (129.5 +/- 89.0 IU/L vs. 94.3 +/- 87.0 IU/L, p = 0.01). Histologic grade was the only factor independently associated with steatosis with multivariate analysis. When compared to the histologic diagnosis of steatosis (n = 122), ultrasonography had a substantial number of false-positive and false-negative results. In patients with a normal ultrasound, 8/82 (10%) had > 30% steatosis on biopsy. Among patients with steatosis reported on ultrasound, only 12/40 (30%) had > 30% steatosis on biopsy review. CONCLUSION: Steatosis on ultrasound is associated with markers of inflammation and fibrosis in HCV-infected patients, but does not consistently correlate with steatosis on biopsy outside of the context of a controlled study. Clinicians should be skeptical of the definitive diagnosis of steatosis on hepatic ultrasonography. PMID- 15829010 TI - Good old days? AB - Alternative models of subsidizing scholarly publishing and dissemination have germinated and gathered momentum in the fertile soil of dissatisfaction. Like the stubborn spring dandelion that needs but a small crack in the sidewalk to flower boldly, the first flowers of Open Access in library literature, including Biomedical Digital Libraries, have sensed their opportunity to change the existing paradigm of giving away our scholarship and intellectual property, only to buy it back for the privilege of knowing it can be read. Will biomedical digital library and informatics researchers understand their role in a new era of Open Access simply by desiring an immediate uninhibited global audience and recognizing the necessity of open access peer-reviewed literature to become self sufficient? PMID- 15829011 TI - Acquisition of anoikis resistance in human osteosarcoma cells does not alter sensitivity to chemotherapeutic agents. AB - BACKGROUND: Chemotherapy-induced cell death can involve the induction of apoptosis. Thus, aberrant function of the pathways involved might result in chemoresistance. Since cell adhesion to the extracellular matrix acts as a survival factor that homeostatically maintains normal tissue architecture, it was tested whether acquisition of resistance to deadhesion-induced apoptosis (anoikis) in human osteosarcoma would result in resistance to chemotherapy. METHODS: Osteosarcoma cell lines (SAOS-2 and TE-85) obtained from ATCC and were maintained in complete Eagle's MEM medium. Suspension culture was established by placing cells in tissue culture wells coated with poly-HEMA. Cell cytotoxicity was determined using a live/dead cytotoxicity assay. Cell cycle/apoptosis analyses were performed using propidium iodide (PI) staining with subsequent FACS analysis. Apoptosis was also assayed by Annexin-FITC/PI staining. RESULTS: Etoposide, adriamycin, vinblastine, cisplatin and paclitaxel were able to induce apoptosis in human osteosarcoma cells SAOS-2 regardless of their anoikis resistance phenotype or the culture conditions (adhered vs. suspended). Moreover, suspended anoikis resistant TE-85 cells (TE-85ar) retained their sensitivity to chemotherapy as well. CONCLUSION: Acquisition of anoikis resistance in human osteosarcoma cells does not result in a generalized resistance to all apoptotic stimuli, including chemotherapy. Moreover, our results suggest that the pathways regulating anoikis resistance and chemotherapy resistance might involve the action of different mediators. PMID- 15829014 TI - Diets and selected lifestyle practices of self-defined adult vegetarians from a population-based sample suggest they are more 'health conscious' AB - BACKGROUND: Few population-based studies of vegetarians have been published. Thus we compared self-reported vegetarians to non-vegetarians in a representative sample of British Columbia (BC) adults, weighted to reflect the BC population. METHODS: Questionnaires, 24-hr recalls and anthropometric measures were completed during in-person interviews with 1817 community-dwelling residents, 19-84 years, recruited using a population-based health registry. Vegetarian status was self defined. ANOVA with age as a covariate was used to analyze continuous variables, and chi-square was used for categorical variables. Supplement intakes were compared using the Mann-Whitney test. RESULTS: Approximately 6% (n = 106) stated that they were vegetarian, and most did not adhere rigidly to a flesh-free diet. Vegetarians were more likely female (71% vs. 49%), single, of low-income status, and tended to be younger. Female vegetarians had lower BMI than non-vegetarians (23.1 +/- 0.7 (mean +/- SE) vs. 25.7 +/- 0.2 kg/m2), and also had lower waist circumference (75.0 +/- 1.5 vs. 79.8 +/- 0.5 cm). Male vegetarians and non vegetarians had similar BMI (25.9 +/- 0.8 vs. 26.7 +/- 0.2 kg/m2) and waist circumference (92.5 +/- 2.3 vs. 91.7 +/- 0.4 cm). Female vegetarians were more physically active (69% vs. 42% active >/=4/wk) while male vegetarians were more likely to use nutritive supplements (71% vs. 51%). Energy intakes were similar, but vegetarians reported higher % energy as carbohydrate (56% vs. 50%), and lower % protein (men only; 13% vs. 17%) or % fat (women only; 27% vs. 33%). Vegetarians had higher fiber, magnesium and potassium intakes. For several other nutrients, differences by vegetarian status differed by gender. The prevalence of inadequate magnesium intake (% below Estimated Average Requirement) was lower in vegetarians than non-vegetarians (15% vs. 34%). Female vegetarians also had a lower prevalence of inadequate thiamin, folate, vitamin B6 and C intakes. Vegetarians were more likely than non-vegetarians to consider various health conditions and food/nutrition concerns when choosing foods. CONCLUSION: In this population-based study, evidence was obtained to indicate that vegetarians appear more 'health conscious' than non-vegetarians, although specific differences were not always consistent by gender. Additional population-based studies are required to determine if the observed gender differences exist in other populations. PMID- 15829013 TI - Health insurance, neighborhood income, and emergency department usage by Utah children 1996-1998. AB - BACKGROUND: It is estimated that approximately half of emergency department (ED) usage in the U.S. and other developed countries is for non-urgent conditions and that this usage is related to availability, social, and economic factors. We examined pediatric ED usage in a U.S. state with respect to income, health insurance status, types of medical conditions, and whether introduction of managed care affected utilization by Medicaid children. METHODS: Emergency department usage rates were calculated from 1996 through 1998 using Utah ED data for children with commercial health insurance, Medicaid, for uninsured children, and by income group estimating neighborhood household income from Zip code of residence. We analyzed usage following the July 1996 transition of Utah Medicaid to managed care. RESULTS: Children with Medicaid had approximately 50% greater ED utilization rates than children with commercial health insurance or uninsured children. The majority of usage for Medicaid and uninsured children was for non traumatic conditions. Only 35% of total ED usage was for non-emergent or non urgent conditions and this was related to both Medicaid and low household income. Children lacking health insurance were more likely to be discharged against medical advice (OR = 2.36, 95% C.I. 1.88-2.96). There was no reduction in Medicaid ED usage following the transition to managed care. CONCLUSION: Usage of ED services is related to both health insurance status and income. Children lacking health insurance and Medicaid children have excessive usage for conditions which could be treated in a primary care setting. That managed care does not reduce Medicaid ED usage is consistent with findings of other studies. PMID- 15829012 TI - Intracellular immunity to HIV-1: newly defined retroviral battles inside infected cells. AB - Studies of the human immunodeficiency virus type 1 (HIV-1) continue to enrich eukaryotic biology and immunology. Recent advances have defined factors that function after viral entry and prevent the replication of proviruses in the infected cell. Some of these attack directly viral structures whereas others edit viral genetic material during reverse transcription. Together, they provide strong and immediate intracellular immunity against incoming pathogens. These processes also offer a tantalizing glimpse at basic cellular mechanisms that might restrict the movement of mobile genetic elements and protect the genome. PMID- 15829015 TI - Titanium dioxide particle-induced goblet cell hyperplasia: association with mast cells and IL-13. AB - BACKGROUND: Inhalation of particles aggravates respiratory symptoms including mucus hypersecretion in patients with chronic airway disease and induces goblet cell hyperplasia (GCH) in experimental animal models. However, the underlying mechanisms remain poorly understood. METHODS: To understand this, the numbers of goblet cells, Muc5ac (+) expressing epithelial cells and IL-13 expressing mast cells were measured in the trachea of sham or TiO2 particles-treated rats using periodic acid-Schiff, toluidine blue and immunohistochemical staining. RT-PCR for Muc-1, 2 and 5ac gene transcripts was done using RNA extracted from the trachea. Differential cell count and IL-13 levels were measured in bronchoalveolar lavage (BAL) fluid. In pretreatment groups, cyclophosphamide (CPA) or dexamethasone (DEX) was given before instillation of TiO2. TiO2 treatment markedly increased Muc5ac mRNA expression, and Muc5ac (+) or PAS (+) epithelial cells 48 h following treatment. RESULTS: The concentration of IL-13 in BAL fluids was higher in TiO2 treated-rats when compared to those in sham rats (p < 0.05). Pretreatment with cyclophosphamide (CPA) decreased the number of neutrophils and eosinophils in BAL fluid of TiO2 treated-rats (p < 0.05), but affected neither the percentage of PAS (+) cells, nor IL-13 levels in the BAL fluids (p > 0.05). In contrast, pretreatment with dexamethasone (DEX) diminished the percentage of PAS (+) cells and the levels of IL-13 (p < 0.05). TiO2 treatment increased the IL-13 (+) mast cells (p < 0.05) in the trachea, which was suppressed by DEX (p < 0.05), but not by CPA pretreatment (p > 0.05). In addition there were significant correlations of IL-13 (+) rate of mast cells in the trachea with IL-13 concentration in BAL fluid (p < 0.01) and with the percentage of Muc5ac (+) cells in the sham and TiO2 treated rats (p < 0.05). CONCLUSION: In conclusion, TiO2 instillation induces GCH and Muc5ac expression, and this process may be associated with increased production of IL-13 by mast cells. PMID- 15829016 TI - Natural health product-HIV drug interactions: a systematic review. AB - The use of natural health products (NHPs) within the HIV community is high. Several NHPs have demonstrated interactions with HIV medications that could contribute to drug failure. We aimed to conduct a systematic review of clinical trials examining NHP-HIV drug interactions and their methodological characteristics. We searched electronic databases and unpublished resources independently, in duplicate. Nine studies were identified, eight clinical pharmacokinetics trials and one population-pharmacokinetics trial. Investigators studied four different herbal medicines (St John's wort, garlic, goldenseal and milk thistle) and one vitamin (vitamin C). Significant interactions were observed with St John's wort, garlic and vitamin C. However, methodological challenges exist to making the results directly generalizable to patients. This review finds that important drug level changes exist when NHPs are combined with HIV medications. Considering patient values and the implications of these studies, further research is urgently required to determine the extent of interactions with other commonly used NHPs. PMID- 15829017 TI - Towards an evidence-based understanding of Peyronie's disease. AB - Aetiopathogenesis and therapy for Peyronie's disease are summarized below. AETIOPATHOGENESIS: (a) Plaque fibroblasts and their products provoke an immune response; (b) there are chromosomal differences between plaques and normal albuginea; (c) plaque fibroblasts are immortalized cells; (d) plaque fibroblasts demonstrate an impaired mitochondrial activity. These patterns provoke inflammation and they compel us to regard Peyronie's disease as being more similar to keloids than to scars. THERAPY: Medical therapy plays a pivotal role in the management of Peyronie's disease and should be performed prior to any surgery. Intraplaque collagenase, intraplaque verapamil, intraplaque interferon, oral acetyl-L-carnitine, oral propionyl-L-carnitine and oral colchicine have proved effective in Peyronie's disease. With the exception of collagenase, these drugs have displayed a number of activities whose final result is to improve immune response, to inhibit inflammation and to inhibit fibroblast metabolism and replication. Therefore, the primitive replicative (rather than reactive) nature of the disease is confirmed; this knowledge may be of help in the identification of new non-surgical therapies for Peyronie's disease. PMID- 15829018 TI - Treatment of category III A prostatitis with zafirlukast: a randomized controlled feasibility study. AB - The cause of category III A prostatitis, chronic prostatitis/chronic male pelvic pain syndrome category A (CP/CPPS A), is uncertain. Treatments for it are based on consensus opinion rather than on scientific data. Our aim was to examine the effect of zafirlukast, a leucotriene antagonist, on the symptoms of CP/CPPS A in our genitourinary (GU) medicine unit. CP/CPPS A was diagnosed by comparative white cell counts of split urine (Stamey) analysis or by finding an excess of polymorphs in expressed prostatic fluid. Symptom change was assessed by the National Institutes of Health Chronic Prostatitis Symptom Index (CPSI). Patients were given zafirlukast or placebo for four weeks in a random double-blind fashion. All patients also received doxycycline. In all, 31 patients were asked to participate and 17 entered the study. No difference in outcome could be shown between the active (10) and placebo (seven) patients. Zafirlukast cannot be demonstrated to be useful in the symptomatic treatment of CP/CPPS A. The problems of recruitment into this study (in spite of a large number of patients with prostatic type pain being seen in our unit) suggest that multicentre treatment trials using non-invasive diagnostic techniques such as the CPSI (rather than single GU medicine units diagnosing CP/CPPS A by uncomfortable direct prostatic testing) are likely to be the most effective and objective methods of undertaking treatment trials in the CP/ CPPS A field in the future. PMID- 15829019 TI - Incidence and presentation of early syphilis diagnosed in HIV-positive gay men attending a central London outpatients' department. AB - We examined the incidence, presentation and sexual behaviour of gay men diagnosed with early syphilis at the Royal Free Hospital HIV department in 2002. A total of 1086 gay men attended and 31 were diagnosed with early syphilis (2.9/100 person years). Twenty-six (84%) of the men were symptomatic and 15 (48%) had documented negative serology within the previous six months. All of the men reported anal intercourse with a new partner in the previous three months. The results support offering regular serological screening to sexually active gay men attending our HIV outpatients' department. PMID- 15829021 TI - A cross-sectional survey of treatment choices for anogenital warts. AB - This was a cross-sectional survey that collected data relating to management of anogenital warts (AGW) during a single-patient visit only at genitourinary medicine clinics. Single-agent use of cryotherapy, podophyllotoxin and trichloroacetic acid (TCA) were the most common treatment modalities, accounting for over two-thirds of all modalities used. Podophyllin, alone or in combination with other agents, was used for about 20% of first-line treatments. Podophyllin was included in about 15% of all treatment modalities. Guidelines for the management of AGW continue to recommend the use of podophyllin, but this may need to be modified in the light of recent publications. Podophyllin, TCA, podophyllotoxin or combinations of these agents are commonly used to treat keratinized warts. About 11% of all treatments involved a combination of two or more agents. PMID- 15829020 TI - How condom use, number of receptive anal intercourse partners and history of external genital warts predict risk for external anal warts. AB - Few analytic opportunities have allowed us to evaluate the role that specific sexual acts and male latex condoms play in the acquisition of external anal warts (EAW) using longitudinal data. The acquisition of EAWs occurs from epithelial contact with other HPV-infected surfaces, and hence is dependent upon sexual behaviour. Our objectives were to classify the relative importance of condom use, receptive anal intercourse (RAI) and prior history of EGWs on acquisition of EAWs. The observational Multicenter AIDS Cohort Study followed 2925 men over nine semiannual study visits for behavioural and physical examinations with laboratory testing. The main outcome measure was the occurrence of examiner-diagnosed EAWs in a homosexual population. EAWs were diagnosed among 10% of men studied across 22,157 visits reviewed for this study. Men with history of EGWs were more likely than those previously unaffected to have developed EAWs (cOR = 2.4 (2.0, 2.9)), as were men who reported multiple anoreceptive intercourse partners (e.g., compared with men who reported no RAI partners, men with 1, 2-5, > or = 6 RAI partners had crude risk ratios 1.0 (0.8, 1.3), 1.6 (1.2, 2.1), 3.9 (2.7, 5.8), respectively). These relations persisted after other demographic and sexual risk factors were controlled for in the analyses. Consistent condom usage showed no protective effect for EAWs in our crude or adjusted analyses. Patient education messages should be tailored to reflect our uncertainty about the protective nature of condoms for the development of anal warts, but to continue to assert the protective effects of a limited lifetime number of sexual partners and the heightened risk for wart recurrence once infected. PMID- 15829022 TI - Use of protective barriers and unprotected sex among adult male prison inmates prior to incarceration. AB - Research documents the challenges associated with preventing HIV/AIDS among prison inmate populations. This paper describes the predictors of protective barrier use and unprotected sexual intercourse among a sample of adult male inmates. Descriptive and scale information was collected on several variables using standard response formats. All major substance use constructs (marijuana, cocaine, and alcohol use) were assessed in terms of consumption prior to incarceration. T-tests were employed to evaluate the level of marijuana, cocaine, and alcohol use in general, as well as with particular respect to sexual situations. Linear regression analysis was conducted to determine whether the use of the substances, length of incarceration, and other demographic variables were associated with the frequency of reporting use of a protective barrier. The findings note that total years in prison, and marijuana and cocaine use were associated with inmates' risk behaviours. Marijuana and cocaine use were statistically significant predictors of risky sex behaviours. PMID- 15829023 TI - Epidemiology and microbiology of subclinical mastitis among HIV-infected women in Malawi. AB - The epidemiology and microbiology of subclinical mastitis, a risk factor for perinatal HIV transmission, have not been well characterized. In all, 250 HIV infected women were followed from two weeks to 12 months postpartum in Blantyre, Malawi, and subclinical mastitis was assessed by breast milk leukocyte counts. The point prevalence of subclinical mastitis at 2, 4, 6, 10, and 14 weeks, and 6, 9, and 12 months was 12.2%, 7.8%, 6.8%, 3.7%, 10.6%, 5.1%, 4.9%, and 1.9%, respectively (P = 0.002), and 27.2% of women had at least one episode of subclinical mastitis. There was no significant relationship between maternal plasma HIV load or parity and subclinical mastitis. Staphylococcus aureus was isolated in 30% of women with subclinical mastitis, and the proportion of women with positive cultures decreased during follow-up (P = 0.02). Subclinical mastitis is prevalent among breastfeeding mothers and further studies are needed to characterize the differences between infectious and non-infectious subclinical mastitis. PMID- 15829024 TI - 'Have a Pap smear!'--doctors, their clients, and opportunistic cervical cancer screening. AB - In many middle-income countries with a high incidence of cervical cancer, organized screening programmes with the Pap test are being planned. We assessed the knowledge of, and attitudes towards, cervical screening among 63 doctors and 102 randomly selected community members in Trinidad where screening is still opportunistic. Doctors were well informed about cervical cancer, but not all knew the approximate specificity of the Pap test. Many did not routinely discuss the benefits and disadvantages of screening with their clients. Most women had heard of the Pap test, but only 56% knew its purpose; 25% would not participate in screening, stating reasons such as being in menopause or not having symptoms. More information about the aim of screening and the purpose of the Pap test must be communicated. Doctors need to keep their knowledge on screening up-to-date, and offer counselling that helps women to make an informed decision whether or not to participate in screening. PMID- 15829025 TI - Sociodemographic factors associated with participation by HIV-1-positive pregnant women in an intervention to prevent mother-to-child transmission of HIV in Cote d'Ivoire. AB - Many HIV-1-seropositive women in Africa who are offered antiretroviral prophylaxis to prevent mother-to-child transmission (MTCT) of HIV do not begin interventions. Research on barriers to participation has not addressed the possible effects of women's sociocultural and economic circumstances. We examined these factors at an MTCT prevention programme in Abidjan, Cote d'Ivoire. We interviewed two groups of women after they had received HIV-positive test results and had been invited by the programme staff to return for monthly follow-up visits before beginning short-course zidovudine prophylaxis. Participants (n = 30) completed follow-up visits and prophylaxis. Non-participants (n = 27) refused or discontinued follow-up visits and did not begin zidovudine. Fewer non participants had been born in Cote d'Ivoire (67% vs. 97%) or were Ivorian nationals (48% vs. 77%); they had lived in the country for less time (21 vs. 26 median years). They were less likely to be French-literate (37% vs. 77%), and more of them reported having had Koranic education only (18% vs. 0). They more often reported miscarriages, stillbirths, or infant deaths (69% vs. 33%), and had partners with low-ranked jobs (63% vs. 30%). Our findings suggest that the non participants were more marginal socioculturally and economically in Ivorian society than participants. Greater attention to mitigating the effects of broader structural factors on women's participation in interventions may increase the effectiveness of MTCT prevention in Africa. PMID- 15829026 TI - Initiation of highly active antiretroviral therapy in advanced AIDS with CD4 < 50 cells/mm3 in a resource-limited setting: efficacy and tolerability. AB - In developing countries, patients often present late with advanced AIDS and a very low CD4 cell count. A retrospective cohort study was conducted in HIV infected patients who had been initiated into highly active antiretroviral therapy (HAART) with CD4 cell count < 50 cells/mm3. There were 159 patients of mean age 36.6 years and 60.4% had previous major opportunistic infections. Mean CD4 was 22 cells/mm3 and 80% had HIV RNA > 100,000 copies/mL. The majority of HAART regimens is non-nucleoside reverse transcriptase inhibitor-based (81.8%). In as-treated analysis, 50, 71.2, 79.7, 79.4, and 80.1% of patients achieved undetectable HIV RNA (< 50 copies/mL) at 12, 24, 36, 48, and 60 weeks, respectively. The corresponding mean CD4 counts were 95, 125, 166, 201, and 225 cells/mm3. Twenty two patients (13.8%) had adverse drug events and half of these had to discontinue HAART. Initiation of HAART in advanced AIDS with CD4 cell count < 50 cells/mm3 is effective, safe, and well tolerated and should not be delayed. PMID- 15829027 TI - Human papillomavirus prevalence at the USA-Mexico border among women 40 years of age and older. AB - The incidence of cervical cancer increases with age among USA Hispanics and women living in Latin America starting in the fourth decade of life. We conducted a study of women > or = 40 living at the USA-Mexico border to determine the prevalence and risk factors for human papillomavirus (HPV) infection detected by polymerase chain reaction. In all, 9.2% of participants tested HPV positive. Compared with women aged 50-59, odds ratios of 8.82 and 6.67 were observed for women > or = 60 and 40-49, respectively. Among women aged 40-49, both oncogenic and non-oncogenic HPV infections were detected; however, women > or = 60 were positive for predominantly oncogenic genotypes. HPV risk significantly increased with > or = 2 lifetime sexual partners in adjusted models. These data suggest that the prevalence of HPV infection may have a second peak among post-menopausal Hispanic women. PMID- 15829028 TI - Study of patients diagnosed with advanced HIV in the HAART era--OMEGA Cohort. AB - Our objective is to analyse patients diagnosed with late-stage HIV infection in the highly active antiretroviral therapy (HAART) area. A prospective, observational study of all patients with an initial CD4 < 50 x 10(6)/L was carried out. Epidemiological, clinical and HAART-associated data were analysed. Survival rates were estimated and pairs of survival curves were compared. The statistical program used was SPSS (version 10). In all, 349 HIV-infected patients were diagnosed, 117 (33.5%) had late-stage disease, mean CD4 23.9 x 10(6)/L and mean viral load (VL) 5.38 log10. In 98 men, mean age 39.5 years, percentage of AIDS cases at their first attendance was 83.8%. The median follow-up period was 28 months and 27 died. Pneumocystis carinii was the most frequent cause of AIDS (24.4%) and death (18.5%). Survival rates at 12, 24 and 36 months were 95.6%, 85.8% and 72.4%. HAART was started in 82.1%. VLs < 50 copies/mL at one, two and three years of treatment were 55.2%, 55.7% and 58.0%. Resource utilization included 0.58 hospitalization/patient/year and 0.07 events/patient/year. HAART related complications were as follows: 50% lipodystrophy, 9.7% hypertension, 22.2% hyperglycaemia, 26.4% hypercholesterolaemia, 31.9% hypertrygliceridaemia and 18.1% mixed hyperlipaemia. Over one-third of our patients have advanced HIV infection at diagnosis. However, the outcome is favourable, with a good immunovirological response and few new opportunistic events. HAART-related complications were frequent. PMID- 15829029 TI - Risk factors for HIV infection among sex workers in Johannesburg, South Africa. AB - Our objective was to determine the prevalence and risk factors for HIV infection among female sex workers in Johannesburg, South Africa. A cross-sectional survey of female sex workers was conducted using interviewer-administered questionnaires. Prevalent sexually transmitted infections including HIV were evaluated through standard laboratory testing. HIV infection was identified in 137 (46.4%) of 295 subjects tested. Increasing frequency of condom use was significantly negatively associated with HIV infection (odds ratio [OR] for moderate use = 0.21; 95% confidence interval [CI]: [0.09, 0.50]; OR for high use = 0.14; 95% CI: [0.06, 0.34]). Sex workers aged > or = 29 years reported significantly different patterns of behaviour than younger workers. Among women aged > or = 29, a negative association with HIV infection (OR = 0.16; 95% CI: [0.07, 0.38]) was found, but only among those not infected with Neisseria gonorrhoeae. Older women in the Johannesburg sex industry may have adaptive behavioural strategies besides condom usage which reduce their risk of acquiring HIV. However, older sex workers with gonorrhoea constitute a high-risk subgroup. PMID- 15829030 TI - HIV and syphilis in young Chinese adults: implications for spread. AB - Our objective was to explore the epidemiology of HIV and syphilis at population level in young Chinese adults. The study was carried out in six areas of three Chinese provinces: Zhejiang, Yunnan and Shaanxi. All couples attending the compulsory premarital examination were recruited. Routinely taken blood was collected onto filter paper. Tests for HIV and syphilis used modified particle agglutination techniques. In all, 17,226 individuals were tested. There were no HIV infections in Zhejiang or Shaanxi. In Yunnan, there were 28 positives: 0.75% (95% confidence interval: 0.5, 1.1), with 22 (78%) in men and 16 (57%) in known high-risk individuals. Syphilis prevalence ranged from 0.33% to 0.71% across study sites, with 58% in men. Although syphilis levels suggest that unsafe sexual activity is occurring in all areas, HIV is not yet affecting the general population of Zhejiang or Shaanxi. The epidemic in Yunnan is being driven by injecting drug users, but there may now be transmission in the general population. PMID- 15829031 TI - A case of linear epidermal naevus presenting as genital warts--a cautionary tale. AB - Linear epidermal naevus (LEN) in the genital area is quite rare. It may present at birth or appear later on in life, in infancy or childhood and occasionally for the first time in adult life. There are several variants of epidermal naevi (EN), which, to the less experienced, can be mistaken for warts. When extensive, it can be associated with abnormalities in other organ systems (epidermal naevus syndrome). The definitive treatment of LEN is surgical ablation with excision of underlying dermis, but this frequently leads to scarring. Laser therapy is an alternative treatment modality and good results have been shown. We report an unusual case of LEN in the genital area in a 60-year-old man presenting as genital warts. PMID- 15829032 TI - Unexpectedly high prevalence of sexually transmitted diseases in married women attending family planning clinics in Yaounde, Cameroon. PMID- 15829035 TI - Journal of Telemedicine and Telecare: expanding horizons. PMID- 15829033 TI - Use of erectile dysfunction medication and unsafe sex among HIV+ men who have sex with men in care. PMID- 15829036 TI - Introduction to the practice of telemedicine. AB - Telemedicine is the delivery of health care and the exchange of health-care information across distances. It is not a technology or a separate or new branch of medicine. Telemedicine episodes may be classified on the basis of: (1) the interaction between the client and the expert (i.e. realtime or prerecorded), and (2) the type of information being transmitted (e.g. text, audio, video). Much of the telemedicine which is now practised is performed in industrialized countries, such as the USA, but there is increasing interest in the use of telemedicine in developing countries. There are basically two conditions under which telemedicine should be considered: (1) when there is no alternative (e.g. in emergencies in remote environments), and (2) when it is better than existing conventional services (e.g. teleradiology for rural hospitals). For example, telemedicine can be expected to improve equity of access to health care, the quality of that care, and the efficiency by which it is delivered. Research in telemedicine increased steadily in the late 1990s, although the quality of the research could be improved--there have been few randomized controlled trials to date. PMID- 15829037 TI - Telemedicine room design. AB - Proper design of rooms for telemedicine store-and-forward and realtime interactive consultations is critical. The challenge is to incorporate communication technology into medical practice to facilitate efficient communication and effective patient care. If this is done properly, the technology becomes unnoticeable to those involved in the telemedicine interaction. This article describes some of the steps that can be taken in a telemedicine room design to achieve this goal. Room location, treatments, background, lighting, power and telecommunications aspects are reviewed. Although every room design will have its own unique challenges, adhering to these basic principles will provide an excellent foundation to begin with. PMID- 15829038 TI - A cost-consequences analysis of minor injuries telemedicine. AB - A cost-consequences analysis of minor injuries telemedicine was performed alongside a randomized controlled trial in a UK peripheral emergency department. The main outcome measures were safety and clinical effectiveness at seven days after presentation. Costs to the National Health Service (NHS) and patients and their families, for 253 patients, were estimated for seven days following randomization. The mean cost to the NHS for the telemedicine patients was 78.61 pounds and for those assessed routinely was 39.15 pounds. For costs incurred by patients and their families the respective figures were 58.24 pounds and 43.95 pounds. Sensitivity analysis showed the initial results to be robust. Telemedicine was a more expensive option for providing minor injuries care in a general-practitioner-supported peripheral emergency department, while consequences did not vary greatly between the different options. PMID- 15829039 TI - Quality and efficacy of a telecolposcopy programme. AB - We tested the performance of telecolposcopy in the diagnosis of various squamous intraepithelial lesions. There were three groups of physicians: Group 1 consisted of 15 gynaecologists who were not trained in colposcopy; Group 2 consisted of six experienced colposcopists (specialists); Group 3 was a supervising team consisting of three experts (colposcopists with more than 15 years of colposcopy experience). The first group took colpophotographs during gynaecological examinations and used a Web browser to compare them with examples from a database of colposcopy findings. Group 2 made their own diagnoses independently. Then the supervising team examined the findings from all 250 patients and made diagnoses that could be considered the 'gold standard'. The findings were identical for Groups 1 and 2 in 219 cases, i.e. an inter-observer agreement of 88%; Cohen's kappa was 0.81. The findings were identical for Groups 1 and 3 in 208 cases, i.e. an inter-observer agreement of 83%; Cohen's kappa was 0.74. The findings were identical for Groups 2 and 3 in 239 cases, i.e. an inter-observer agreement of 96%; Cohen's kappa was 0.93. There was high inter-observer agreement between all participants. However, there were also significant differences in some cases. In the cases of suspected invasive carcinoma, the results showed a significant difference (chi2=4.8, P< 0.005) because gynaecologists from Group 1 were not sure of their diagnosis and they wanted to obtain a second opinion by referring the decision to a higher level. PMID- 15829040 TI - Publication output in telemedicine in Spain. AB - We searched the MEDLINE database for publications about telemedicine from 1966 to 2003. Only publications with a first institutional address in Spain and from journals listed in the JCR Science Edition 2002 were included in the study. A total of 118 publications were found. The first publication appeared in 1988 and publication output showed a sudden increase starting in 1995, reaching a maximum in 2002 with 21 publications. The number of authors per publication in non multicentre studies ranged from 1 to 14 (median 6.5). Most authors (79%) contributed to only a single publication; 10% of authors participated in three or more publications. Most publications were full papers (88%). The most active Autonomous Community in the number of publications relative to inhabitants was Galicia (0.91 publications per 100,000 inhabitants). The Community that produced the highest absolute number of publications was Madrid (29% of the total). The mean impact factor of the journals in which the publications appeared was 0.961. Production of telemedicine publications in Spain has followed the same temporal course as has been observed worldwide. There is an unequal geographical and institutional distribution of publications. The highest production is concentrated in a few institutions and only a small number of authors show steady research activity. PMID- 15829041 TI - A telemedicine survey among Milan doctors. AB - We surveyed physicians' attitudes to the use of telemedicine. A questionnaire was sent to 2987 Italian physicians and 361 responded (12%). The topics covered by the survey were: current telemedicine technology knowledge/usage level; general attitude towards telemedicine applications; perceived efficacy of telemedicine in enhancing quality of care; and intention to use telemedicine. In all, 83% of respondents had heard about telemedicine. Some doctors considered telemedicine to be of limited interest and many physicians were not convinced that it could improve clinical practice. These beliefs were more common among doctors of higher seniority, probably because they are more reluctant to accept change. To reduce such negative evaluations, better dissemination of information about the state of the art of research and development in telemedicine is needed. A logistic regression model was used to determine which factors influenced the intention to use telemedicine. The final model predicted 80% of the responses correctly. PMID- 15829042 TI - An assessment of the telehealth needs and health-care priorities of Tanna Island: a remote, under-served and vulnerable population. AB - We surveyed eight Canadian physicians who had each provided medical care for six months on the remote and under-served island of Tanna in Vanuatu. The most frequently encountered medical problems on Tanna were infectious diseases (tuberculosis, hepatitis, abscesses, malaria, pneumonia, typhoid fever, meningitis and skin infections). When physicians were asked about the top three health-care priorities, they ranked tuberculosis control, clean water and improved health-care delivery/communication between hospital and outposts as most important. The key issues were: (1) basic public health needs and infrastructure development are higher in priority than telehealth; (2) telehealth consultants must have knowledge pertinent to local conditions and resources available to the population; (3) electronic equipment suited to tropical environments is needed; (4) projects must be developed locally rather than internationally. Understanding how telehealth can provide support to health professionals under challenging conditions may assist with the health priorities in developing countries and potentially provide access to resources both locally and internationally. PMID- 15829043 TI - Pre-hospital diagnosis of myocardial ischaemia by telecardiology: safety and efficacy of a 12-lead electrocardiogram, recorded and transmitted by the patient. AB - We compared a 12-lead electrocardiogram (ECG) recorded by the patient and transmitted to a cardiology call centre via telephone (tele-ECG) with a standard 12-lead ECG recorded from the same patient at the same time. In 158 patients, tele-ECGs were compared with standard ECGs by two cardiologists and one internist, independently and blindly. In 14 patients peripheral electrodes were displaced, and in 12 patients there were baseline artefacts. These technical errors were corrected by telephone communication in all but two individuals. One patient could not use the tele-ECG device because of disability. Hence, in 155 of 158 patients (98%), the quality of the tele-ECG was adequate for diagnosis. Reliability coefficients (R) for PQ, QRS and QT intervals between tele- and standard ECG were high, with R values of 0.73, 0.75 and 0.79, depending on the physician. Negative T-waves could be detected with very good agreement in the tele-ECG as compared with the standard ECG (kappa values of 0.97, 0.95 and 0.94). The agreement between tele- and standard ECG concerning alterations of the ST segment was very good (kappa =0.99 for all investigators). Residual signs of myocardial infarction could be detected by tele-ECG, with very good agreement for anterior as well as for posterior localizations (kappa =0.99 and 1.00). The tele ECG technique seems a promising approach to reducing pre- and in-hospital time delays to the initiation of thrombolytic therapy. PMID- 15829044 TI - Organizational cooperation in teleradiology. AB - Teleradiology requires cooperation between participating parties, but it cannot be assumed that such cooperation will work well. A survey was carried out of 12 radiology departments in Norway which used picture archiving and communication systems in connection with teleradiology. Qualitative interviews were carried out with 26 out of 29 resource persons (response rate 90%). The respondents identified 17 issues of importance for cooperation in teleradiology. None of the problems with cooperation seem large enough to prevent effective teleradiology cooperation in future. For organizations planning teleradiology of a larger volume, the cooperation issue is important. It is recommended that managers lead change processes in their organizations where the different issues of importance to cooperation are treated and the right measures are taken to realize the full potential of teleradiology. For telemedicine, it is important that future research includes investigations on cooperation for the different applications. PMID- 15829046 TI - How 'colour blind' is telemedicine? PMID- 15829048 TI - Teleneurology. AB - Teleneurology enables neurology to be practised when the doctor and patient are not present in the same place, and possibly not at the same time. The two main techniques are: (1) videoconferencing, which enables communication between a doctor and a patient who are in different places at the same time (often called realtime or synchronous), and (2) email, where the consultation is carried out without the patient being present, at a time convenient to the doctors involved (asynchronous or store-and-forward teleneurology). Some problems that can be solved by teleneurology include: (1) patients admitted to hospital with acute neurological symptoms rarely see a neurologist; (2) delayed treatment for acute stroke; (3) non-optimum management of epilepsy; (4) unproductive travel time for neurologists; (5) extremely poor access to a neurologist for doctors in the developing world; (6) long waiting times to see a neurologist. Neurology is a specialty that, because of the emphasis on accurate interpretation of a history, does lend itself to telemedicine. It has been a late starter in realizing the benefits of telemedicine and most of the publications on teleneurology have been in the last five years. Its uptake within the neurological community is low but increasing. Telemedicine requires a significant change in how neurologists practise. The evidence to date is that teleneurology can narrow the gap between patients with neurological disease and the doctors who are trained to look after them. PMID- 15829049 TI - Benefits and drawbacks of telemedicine. AB - Telemedicine is a vast subject, but as yet there are limited data on the clinical effectiveness and cost-effectiveness of most telemedicine applications. As a result, objective information about the benefits and drawbacks of telemedicine is limited. This review is therefore based mainly on preliminary results, opinions and predictions. Many potential benefits of telemedicine can be envisaged, including: improved access to information; provision of care not previously deliverable; improved access to services and increasing care delivery; improved professional education; quality control of screening programmes; and reduced health-care costs. Although telemedicine clearly has a wide range of potential benefits, it also has some disadvantages. The main ones that can be envisaged are: a breakdown in the relationship between health professional and patient; a breakdown in the relationship between health professionals; issues concerning the quality of health information; and organizational and bureaucratic difficulties. On balance, the benefits of telemedicine are substantial, assuming that more research will reduce or eliminate the obvious drawbacks. PMID- 15829050 TI - Network basics for telemedicine. AB - Early telemedicine networks employed dedicated telecommunications circuits (e.g. leased digital lines) in which the sender and receiver were connected by a private circuit. More recently, the Internet has become widely available for general use, including telemedicine. The Internet was engineered to permit network paths to be shared by all users, so data transmission is fundamentally different from traditional, circuit-switched networks. Early telemedicine applications demonstrated the feasibility of Internet Protocol transmission. The basic performance criteria to use in evaluating newer digital communications technologies that carry both voice and data are: (1) bandwidth; (2) packet loss; (3) end-to-end delay; (4) jitter; (5) privacy and security. Network engineering involves performance trade-offs between the hardware, architecture, security and the budget available. A telemedicine application may be running over a network whose design is entirely under the user's control, or the application may employ some part of the Internet whose design is unknown to the user. If an application is not running to satisfaction, then a network engineer should be consulted. PMID- 15829051 TI - An incremental cost analysis of telehealth in Nova Scotia from a societal perspective. AB - We examined the costs of telehealth in Nova Scotia from a societal perspective. The clinical outcomes of telepsychiatry and teledermatology services were assumed to be similar to those for conventional face-to-face consultations. Cost information was obtained from the Nova Scotia Department of Health, the Canadian Institute for Health Information, and questionnaires to patients, physicians and telehealth coordinators. There were 215 questionnaires completed by patients, 135 by specialist physicians and eight by telehealth coordinators. Patient costs for a face-to-face consultation ranged from $240 to $1048 (all costs in Canadian dollars), whereas patient costs for telehealth were lower, from $17 to $70. However, from a societal perspective, the overall cost of providing face-to-face services was lower than for telehealth: the total costs for face-to-face services ranged from $325 to $1133, while the total costs for telehealth services ranged from $1736 to $28,084. A threshold analysis showed that, above a certain patient workload, telehealth services would be more cost-effective than face-to-face services from a societal perspective. This workload is attainable in Nova Scotia. PMID- 15829052 TI - Coordinated care planning for elderly patients using videoconferencing. AB - We studied the transfer of information during coordinated care planning between a university hospital and a local health care centre/social welfare department about 35 km away. During a seven-month study period, 10 sessions were conducted by videoconferencing and seven sessions were conducted by face-to-face conferencing. Videoconferencing reduced the time required for each coordinated care-planning session from an average of 60 to 45 min. There was also an increase in the number of participating professional categories. Travel time for the staff in the face-to-face group was 60-180 min each. Use of a care-planning report during the sessions resulted in improved quality of documentation, which contributed to better care following discharge. The technical problems that occurred did not detract from the beneficial experience of participating. Interviews with next of kin showed that they had been able to influence the content of the care during the care-planning sessions. Videoconferencing proved useful in coordinated care planning. It resulted in time saved due to reduced travel time, participation by more staff categories and an enhancement of the documentation quality. PMID- 15829053 TI - Online diabetes education: design and evaluation with prospective diabetes camp counsellors. AB - An educational Website was designed by the Florida Initiative in Telehealth and Education group, and an online diabetes education test was developed using a sample of 60 children and young adults aged 8-22 years, all of whom had diabetes. The 31 items were analysed for item difficulty. Eight test items were eliminated as being unsuitable. The test was then used in 67 prospective diabetes counsellors (23 men, 44 women) who volunteered for a summer camp. Camp counsellors ranged in age from 17 to 33 years (mean 22 years, SD 3). The counsellors' mean pre-test scores were 80% and their mean post-test scores were 92%. There was a significant improvement (P=0.001) of approximately 1.25 questions from pre- to post-test. This supports the use of the online educational Website for training individuals working with children with diabetes. The Website may prove to be useful for online education in other areas of diabetes management. PMID- 15829054 TI - A comparison of face-to-face versus remote assessment of neonatal resuscitation skills. AB - The neonatal resuscitation skills of 30 third-year medical students were assessed in real time by a face-to-face examiner in the same room as the student, and by a remote examiner located in a separate room using the ANAKIN system. The ANAKIN system combines an instrumented manikin simulator, computer-based assessment and high-bandwidth videoconferencing. The students were assessed while performing a neonatal resuscitation megacode using the ANAKIN system. Students were satisfied with the ANAKIN system as an assessment system and were not intimidated by its use. However, the correlation between the face-to-face and remote examiner's mean total performance assessment scores was 0.27, which was not significant (P=0.14). The results indicated variation between the examiners' performance scores in a number of key technical skill areas. The findings from this study have implications for the use of technology-mediated systems in assessing resuscitation skills. Examiner orientation is critical for individuals using such systems. These persons must be comfortable and confident in using the technology. Interface and design features of the system need to be carefully scrutinized and tested. PMID- 15829055 TI - Evaluation of a digital store-and-forward colposcopic system--a pilot study to assess usability for telemedicine. AB - We evaluated the diagnostic reliability of cervical examination using digital colposcopy compared with conventional binocular colposcopy. A total of 315 patients were examined and diagnosed by a colposcopist on-site. During the colposcopic examination, digital camera images were stored on a PC (median five pictures per patient). A second physician, experienced in colposcopy, re evaluated the initial diagnostic findings using the stored digital images. The primary and secondary findings in each patient were categorized according to the Rome classification system. There was agreement between the primary and secondary examiners in 69% of cases (kappa=0.60). There was no systematic bias in terms of under- or over-rating. The proportion of non-assessable colposcopic examinations was 9%. Digital colposcopy was reliable and provided advantages in terms of a better follow-up examination and internal quality control of the diagnosis. The pilot study suggests that telecolposcopy may provide better training and further education for physicians and students, and may also improve the diagnostic possibilities in gynaecology. PMID- 15829058 TI - Cupular deposits and aminoglycoside administration in human temporal bones. AB - In this study, the deposits of basophilic material on the cupula of the semicircular canals in temporal bones from patients who had aminoglycoside administration within six months prior to death were compared with normal temporal bones. Subjects were divided into two groups. Group I included 24 normal control temporal bones age-matched to group II patients. Group II consisted of 23 temporal bones that had received aminoglycosides within six months prior to death. All temporal bones were examined under light microscopy. One (4.2 per cent) of 24 temporal bones in group I (normal) showed basophilic deposits. In group II, deposits were observed in 8 (34.8 per cent) of 23 temporal bones. The prevalence of basophilic deposits in group II was significantly higher than group I. This study demonstrates that within six months after aminoglycoside administration there is an increased prevalence of basophilic deposits on the surface of the cupula. Such changes may be related to the benign paroxysmal positional vertigo (BPPV) seen in some patients who have had aminoglycoside administration. PMID- 15829059 TI - Targeted therapy of human laryngeal squamous cell carcinoma in vitro by antisense oligonucleotides directed against telomerase reverse transcriptase mRNA. AB - A number of different approaches have been developed to inhibit telomerase activity in human cancer cells. In this study, the effect of antisense oligonucleotides (ODNs) by targeting human telomerase reverse transcriptase (hTERT) mRNA in a laryngeal cancer cell line (Hep-2) was investigated. A 20mer antisense oligodeoxynucleotide targeting the most open part of hTERT mRNA (anti hTERT) and a mismatched control sequence were synthesized. Cells were treated daily with oligonucleotides for up to 72 hours. hTERT mRNA expression was measured by the reverse transcription polymerase chain reaction (RT-PCR) assay; telomerase activity by the telomerase PCR ELISA assay kit (TRAP; Boehringer Mannheim, GmbH, Mannheim, Germany). Cell viability after administration of ODNs was determined using the MTT assay. Morphological changes were examined by haematoxylin and eosin staining. The cell cycle was analyzed using flow cytometry. It was found that antisense treatment induced a decrease in hTERT mRNA expression, telomerase activity, cell growth rate, cell viability, and an increase in apoptosis. The results suggest that inhibition of telomerase activity in Hep-2 cells by short-term antisense treatment against the mRNA of hTERT results in apoptotic cell death. The treatment with anti-hTERT may be useful as a treatment modality for laryngeal squamous carcinoma. PMID- 15829060 TI - BAHNO surgical specialities: same patients, different practices? AB - The British Association of Head and Neck Oncologists (BAHNO) is 'a multidisciplinary society for healthcare professionals involved in the study and treatment of head and neck cancer'. Surgical members of this organization are from three specialities (otolaryngology, maxillo-facial and plastic surgery). Although the overall impression is that the management of UK head and neck cancer patients is consensus based, there are appreciable differences in each surgical speciality's practice. Anecdotally, this can lead to variation in the management of very similar patients. To identify some of these variations BAHNO surgeons were surveyed regarding their current head and neck cancer practices from the perspectives of surgical activity and post-operative care. Some unexpected differences were identified, particularly in relation to post-operative care with plastic and maxillo-facial surgeons demonstrating different patterns of high dependency unit (HDU) and intensive care unit (ICU) use for the same patients. The implications for future consensus in the light of these variations are discussed. PMID- 15829061 TI - A national survey amongst UK otolaryngologists regarding the treatment of Meniere's disease. AB - Since Dr Prosper Meniere described the vertiginous syndrome that now bears his name, a large variety of medical and surgical treatments have been introduced. To determine the way in which this condition is currently managed in the United Kingdom, a postal survey amongst consultant otolaryngologists was carried out. It revealed that 52 per cent were actively involved in the treatment of patients with Menieres disease using a wide range of medical and surgical therapies that have little or no evidence base. The survey found that 94 per cent of surgeons prescribe betahistine, 63 per cent diuretics and 71 per cent advise salt restriction to their patients, while 52 per cent of surgeons continue to recommend saccus decompression and 50 per cent are still inserting a grommet. However, two thirds of respondents now advocate the use of gentamicin therapy despite it only being introduced to this country just over 10 years ago. The results of this study and their relevance to the recommended present day management of Meniere's disease are discussed. PMID- 15829062 TI - Sleep nasendoscopy as a predictor of treatment success in snorers using mandibular advancement splints. AB - This study assessed the effect on the upper airway during sleep nasendoscopy of mimicking the action of a mandibular advancement splint. Twenty-seven subjects with a diagnosis of sleep-disordered breathing were referred for mandibular advancement splint therapy following sleep nasendoscopy. Sleep nasendoscopy was repeated for all subjects with, and without, the appliance in situ. Follow-up sleep studies with a mandibular advancement splint in situ were undertaken for 19 individuals with significant obstructive sleep apnoea. With the mandibular advancement splint, subjective snoring levels and airway patency improved as predicted in all but one individual. Residual palatal flutter was predicted for five subjects and occurred in eight individuals. Follow-up sleep studies showed highly statistically significant reductions in median apnoea-hypopnoea index (from 28.1 to 6.1, p < 0.001). Mimicking the action of a mandibular advancement splint during sleep nasendoscopy helps considerably in the patient selection process for this form of treatment. PMID- 15829063 TI - Course and long-term outcome of 'refractory' secretory otitis media. AB - OBJECTIVE: The course and the long-term outcome of 'refractory' secretory otitis media (SOM), defined as continuous SOM for more than 6 years, were studied in 52 young patients. They had during childhood been treated for bilateral SOM on average for 12 years (range 6 to 26 years). The mean interval between resolution of SOM and examination was 7 years. METHODS: At follow up the patients' medical records were scrutinized with regards to transmyringeal ventilation tubes, adenoidectomy, sequelae and complications, and a questionnaire was filled in to document other diseases. RESULTS: The onset of SOM showed two peaks, one at the age of one year and one at the age of 3.5 years. Patients whose onset of SOM was related to an episode of acute otitis media (AOM) were younger at SOM onset than those who had no such relation. Otorrhoea and AOM episodes were more frequent during the SOM periods, with blocked or expelled tubes, than during periods with patent tubes. Cholesteatoma were seen in 3 per cent and perforations in 5 per cent of patients. CONCLUSIONS: Extremely long-standing SOM does not necessarily result in myringeal perforation or cholesteatoma to a greater extent than that seen in patients with shorter durations of SOM. However, at follow up one-third of the patients reported hearing impairment and a majority felt discomfort when flying or diving. PMID- 15829064 TI - Nasal fibrosis: long-term follow up of four cases of eosinophilic angiocentric fibrosis. AB - Eosinophilic angiocentric fibrosis is a rare, benign cause of submucosal thickening and fibrosis within the upper respiratory tract. It predominantly affects the nose although cases have been reported in the subglottis. We describe four cases of the disease centred around the nasal cavity, with widespread infiltration of the facial soft tissues and orbit in three of the four patients. Each underwent long term follow up. Multiple surgical resections were required with two of our patients and, to date, medical therapy has been of limited help. The disease process, with its clinical and characteristic histopathological findings, is described. We also discuss the management of the disease following a comprehensive review of, and comparison with, the few prior reported cases. PMID- 15829065 TI - Use of the automatic tympanometer as a screening tool for congenital choanal atresia. AB - Choanal atresia is a significant congenital anomaly, which may be associated with other congenital anomalies. Recommendations have been previously proposed for neonatal screening of all newborns to detect this condition. A previously unreported, simple method using the automatic tympanometer to screen newborns for congenital choanal atresia is proposed. A prospective study was performed over a one-month period on all newborns at El-Sahel Teaching Hospital, Cairo, to evaluate the efficacy of this manoeuvre. The nostrils of 143 newborn infants were evaluated during the study period. The sensitivity of the technique in diagnosing a patent nostril was 100 per cent. The specificity of the technique for detecting distal obstruction was 100 per cent. There were no complications of the described procedures. The study revealed that the novel method of screening for choanal atresia in newborns using the automatic tympanometer is simple, minimally invasive and highly reproducible. PMID- 15829066 TI - An innovation in insert visual reinforcement audiometry in children. AB - Visual reinforcement audiometry (VRA) is an important behavioural test of hearing in young children. The use of insert earphones during VRA provides a reliable method of obtaining ear- and frequency-specific data. Two testers are usually required for this procedure. The first tester operates the audiometer and other controls while the second tester distracts the child with various toys. The paper describes a technique of providing a visual cue to the second tester when a sound stimulus is introduced through the insert earphones. PMID- 15829067 TI - Direct phonoplasty under local anaesthetic. AB - OBJECTIVE: A new technique of direct phonoplasty under local anaesthetic is described. BACKGROUND: Vocal-fold medialization is an accepted method to improve voice in patients with a paralysed vocal fold. Various materials have been used to inject into the paralysed vocal fold, resulting in medialization of the fold and improved phonation. The two established methods are direct injection under general anaesthetic and transcutaneous injection under local anaesthetic; both have advantages and disadvantages. METHOD: A Pentax 80K series flexible endoscope is used to directly inject collagen into the paralysed vocal fold via a reusable 23G endoscopic needle under local anaesthetic. CONCLUSION: New technology and innovation have allowed development of the new technique of direct phonoplasty under local anaesthetic. This combines the advantages of transcutaneous injection under local anaesthetic and direct injection under general anaesthetic, without their disadvantages. PMID- 15829068 TI - Vocal fold palsy due to plombage for tuberculosis. AB - A 67-year-old patient, who had previously undergone Lucite ball plombage for pulmonary tuberculosis, presented with a hoarse voice, intermittent stridor and breathlessness. Direct laryngoscopy confirmed a left vocal fold palsy. A left supraclavicular mass became apparent and a computerized tomograph (CT) scan showed that a Lucite ball had migrated into her supraclavicular fossa. Subsequently she developed left arm pain and weakness. The balls were removed surgically, following which her arm symptoms improved but her voice remained unchanged. Migration of implanted material should be considered when new symptoms appear in patients who have undergone plombage treatment. PMID- 15829069 TI - Parasellar epidermoid cyst rupturing into the nasopharynx. AB - Epidermoid tumours are non-neoplastic inclusion cysts representing up to 1.1 per cent of all intracranial tumours, typically presenting with symptoms related to pressure or intracranial rupture in the fourth or fifth decade of life. The authors present a case of a parasellar epidermoid cyst which has ruptured in to the nasopharynx; to the best of their knowledge, this has not been previously reported. The computed tomography (CT) and magnetic resonance imaging (MRI) are presented. The pathology and radiological features of epidermoid tumours are discussed, particularly in relation to extracranial connections. The differential diagnosis of lesions eroding the central skull base is reviewed. PMID- 15829070 TI - Bilateral traumatic facial paralysis associated with unilateral abducens palsy: a case report. AB - Bilateral traumatic facial paralysis is a very rare clinical condition. Abducens palsy, associated with bilateral traumatic paralysis, is even rarer and has not been well described in the literature. In this report, a 24-year-old male, who developed immediate bilateral facial and right abducens paralyses following a motor vehicle accident, is presented. The patient was referred for neurotologic evaluation 22 days after the injury. Electroneurography (ENoG) demonstrated 100 per cent degeneration at the first examination and, correspondingly, electromyography showed no regeneration potentials. Using high-resolution computed tomography (HRCT), a longitudinal fracture on the right and a mixed-type fracture on the left were identified. The patient had good cochlear reserve on both sides. The decision for surgery was based not on ENoG, because of the delayed referral of the patient, but on the HRCT, which showed clear fracture lines on both sides. The middle cranial fossa approach for decompression of the right facial nerve was performed on the 55th day following the trauma, and a combined procedure using the middle cranial fossa and transmastoid approaches was applied for decompression of the left facial nerve on the 75th day following the trauma. On the right, there was dense fibrosis surrounding the geniculate ganglion and the proximal tympanic segment whereas, on the left, bone fragments impinging on the geniculate ganglion, dense fibrosis surrounding the geniculate ganglion, and a less extensive fibrotic tissue surrounding the pyramidal segment were encountered. There were no complications or hearing deterioration. At the one-year follow up, the patient had House-Brackmann (HB) grade 1 recovery on the right, and HB grade 2 recovery on the left side, and the abducens palsy regressed spontaneously. The middle cranial fossa approach and its combinations can be performed safely in bilateral temporal bone fractures as labyrinthine sparing procedures if done on separate occasions. PMID- 15829071 TI - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) presenting with sudden sensorineural hearing loss. AB - CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is an autosomal dominant angiopathy characterized by recurrent cerebrovascular events, migraine and dementia. We describe a case of sensorineural hearing loss as the presenting feature of this condition. We have found no previous reports in the world literature of CADASIL presenting with a sudden sensorineural hearing loss. The significance of questioning a patient with regard to family history is exemplified in this case. PMID- 15829072 TI - Simultaneous antrochoanal and sphenochoanal polyps: a rare clinical entity. AB - Choanal polyps almost always appear as solitary growths and most commonly arise from the maxillary sinus. Sphenochoanal and ethmoidochoanal polyps are extremely rare. Co-existence of more than one choanal polyp is even more infrequent. We present a patient with an antrochoanal and an accompanying sphenochoanal polyp for the first time in the literature. We discuss the clinical presentation, pathogenesis and surgical management of this rare clinical entity. PMID- 15829073 TI - Parapharyngeal metastasis from papillary thyroid carcinoma: a case diagnosed by thyroglobulin measurement in peroral fine-needle aspiration of a cystic metastatic lymph node. AB - We report a case of a parapharyngeal cystic metastatic lymph node arising from papillary thyroid carcinoma (PTC). Parapharyngeal metastases arising from PTC are rare and correct diagnosis of the parapharyngeal mass before surgery is difficult. In this case, the diagnosis of a parapharyngeal mass was made pre operatively by thyroglobulin measurement in peroral fine-needle aspiration with negative cytology. PMID- 15829074 TI - Proboscis lateralis: clinical and radiological features. AB - Proboscis lateralis is one of the rare craniofacial congenital anomalies which presents as an obvious deformity of the nose. It tends to occur in male offspring of consanguineous marriages. This is a report of computerized tomography of this rare anomaly. In proboscis lateralis, the nasal cavity on one side is completely normal, while on the affected side it is replaced by a trunk-like process attached to the medial portion of the orbital roof. This anomaly occurs sporadically as an isolated defect or in association with other anomalies. It is usually associated with developmental failure of the paranasal sinuses and the nasolacrimal duct. PMID- 15829075 TI - Kerion: an unusual presentation in the otolaryngology department. AB - A 19-year-old farmer was referred by his general practitioner as an emergency to our otolaryngology department complaining of marked breathlessness of a few hours duration. He gave a three-day history of painful swelling and hair loss in the beard area of the right side of the neck. His upper airway was compromised unless extension of the neck was maintained. Larynx and pharynx were normal. The acute symptoms settled with intravenous antibiotics and hydrocortisone. Culture of skin scrapings revealed a growth of Tricophyton verrucosum. The neck swelling subsided after a course of oral griseofulvin followed by terbinafine. Difficulty in breathing due to fungal infection of the neck has not been previously reported in the English literature. PMID- 15829076 TI - Evidence for multiple, distinct representations of the human body. AB - Previous data from single-case and small group studies have suggested distinctions among structural, conceptual, and online sensorimotor representations of the human body. We developed a battery of tasks to further examine the prevalence and anatomic substrates of these body representations. The battery was administered to 70 stroke patients. Fifty-one percent of the patients were impaired relative to controls on at least one body representation measure. Further, principal components analysis of the patient data as well as direct comparisons of patient and control performance suggested a triple dissociation between measures of the 3 putative body representations. Consistent with previous distinctions between the "what" and "how" pathways, lesions of the left temporal lobe were most consistently associated with impaired performance on tasks assessing knowledge of the shape or lexical-semantic information about the body, whereas lesions of the dorsolateral frontal and parietal regions resulted in impaired performance on tasks requiring on-line coding of body posture. PMID- 15829077 TI - Behavioral change and its neural correlates in visual agnosia after expertise training. AB - Agnosia, the impairment in object and face recognition despite intact vision and intelligence, is one of the most intriguing and debilitating neuropsychological deficits. The goal of this study was to determine whether S.M., an individual with longstanding visual agnosia and concomitant prosopagnosia, can be retrained to perform visual object recognition and, if so, what neural substrates mediate this reacquisition. Additionally, of interest is the extent to which training on one type of visual stimulus generalizes to other visual stimuli, as this informs our understanding of the organization of ventral visual cortex. Greebles were chosen as the stimuli for retraining given that, in neurologically normal individuals, these stimuli can engage the fusiform face area. Posttraining, S.M. showed significant improvement in recognizing Greebles, although he did not attain normal levels of performance. He was also able to recognize untrained Greebles and showed improvement in recognizing common objects. Surprisingly, his performance on face recognition, albeit poor initially, was even more impaired following training. A comparison of pre- and postintervention functional neuroimaging data mirrored the behavioral findings: Face-selective voxels in the fusiform gyrus prior to training were no longer so and were, in fact, more Greeble-selective. The findings indicate potential for experience-dependent dynamic reorganization in agnosia with the possibility that residual neural tissue, with limited capacity, will compete for representations. PMID- 15829078 TI - Development of sensitivity to texture and contour information in the human infant. AB - Texture discrimination and bounding contour extraction are essential parts of the object segmentation and shape discrimination process. As such, successful texture and contour processing are key components underlying the development of the perception of both objects and surfaces. By recording visual-evoked potentials, we investigate whether young infants can detect orientation-defined textures and contours. We measured responses to an organized texture comprised of many Gabor patches of the same orientation, alternated with images containing the same number of patches, but all of random orientation. These responses were compared with a control condition consisting of the alternation between two independently random configurations. Significant difference potentials were found as early as 2 5 months, as were significant odd harmonics in the test conditions. Responses were also measured to Gabor patches organized either as circles (all patches tangent to an imaginary circular path) alternated with pinwheels (all patches having a fixed orientation offset from the path). Infants between 6 and 13 months also showed sensitivity to the global organization of the elements along contours. Differential responses to our texture and contour stimuli and their controls could only have been generated by mechanisms that are capable of comparing the relative orientation of 2 or more patches, as no local information at a single patch distinguished the random and organized textures or the circle and pinwheel configurations. PMID- 15829079 TI - Partially distributed representations of objects and faces in ventral temporal cortex. AB - Object and face representations in ventral temporal (VT) cortex were investigated by combining object confusability data from a computational model of object classification with neural response confusability data from a functional neuroimaging experiment. A pattern-based classification algorithm learned to categorize individual brain maps according to the object category being viewed by the subject. An identical algorithm learned to classify an image-based, view dependent representation of the stimuli. High correlations were found between the confusability of object categories and the confusability of brain activity maps. This occurred even with the inclusion of multiple views of objects, and when the object classification model was tested with high spatial frequency "line drawings" of the stimuli. Consistent with a distributed representation of objects in VT cortex, the data indicate that object categories with shared image-based attributes have shared neural structure. PMID- 15829080 TI - To look or not to look? Typical and atypical development of oculomotor control. AB - The ability to inhibit saccades toward suddenly appearing peripheral stimuli (prosaccades) and direct them to contralateral locations instead (antisaccades) is a crucial marker of eye movement control. Typically developing infants as young as 4-month-olds can learn to inhibit reflexive saccades to peripheral stimuli, but they do not produce antisaccades, whose development later in infancy and its underlying neural computations remain unexplored. Here we tested oculomotor control in typically developing toddlers and toddlers with fragile X syndrome (FXS), a disorder of known genetic origin that allows the investigation of the neuro-computational properties contributing to the development of saccadic control. Typically developing toddlers decreased looking toward peripheral cues that predicted contralateral rewards, whose appearance they anticipated. Furthermore, this correlated with age, indicating a gradual development of saccadic control. In contrast with the typical case, toddlers with FXS did not decrease their looks to peripheral onsets that predicted contralateral events. Importantly, the atypical pattern of performance was also evident in the elimination of the correlation with mental or chronological age found in controls. Taken together, the findings suggest that control of saccades and its developmental trajectory is atypical in toddlers with FXS, consistent with inhibitory deficits previously shown at later ages in this condition. Potential implications for the neural mechanisms underlying the typical and atypical development of oculomotor control are discussed. PMID- 15829081 TI - An event-related potential study of selective auditory attention in children and adults. AB - In a dichotic listening paradigm, event-related potentials (ERPs) were recorded to linguistic and nonlinguistic probe stimuli embedded in 2 different narrative contexts as they were either attended or unattended. In adults, the typical N1 attention effect was observed for both types of probes: Probes superimposed on the attended narrative elicited an enhanced negativity compared to the same probes when unattended. Overall, this sustained attention effect was greater over medial and left lateral sites, but was more posteriorly distributed and of longer duration for linguistic as compared to nonlinguistic probes. In contrast, in 6- to 8-year-old children the ERPs were morphologically dissimilar to those elicited in adults and children displayed a greater positivity to both types of probe stimuli when embedded in the attended as compared to the unattended narrative. Although both adults and children showed attention effects beginning at about 100 msec, only adults displayed left-lateralized attention effects and a distinct, posterior distribution for linguistic probes. These results suggest that the attentional networks indexed by this task continue to develop beyond the age of 8 years. PMID- 15829082 TI - Common mechanisms for working memory and attention: the case of perseveration with visible solutions. AB - Everyone perseverates at one time or another, repeating previous behaviors when they are no longer appropriate. Such perseveration often occurs in situations with working memory demands, and the ability to overcome perseveration has been linked to brain regions critical for working memory. Many theories thus explain perseveration in terms of working memory deficits. However, perseveration also occurs in situations without apparent working memory demands, in which the visible environment specifies appropriate behavior. Such findings appear to challenge working memory accounts of perseveration. To evaluate this challenge, a neural network model of a working memory account of perseveration was tested on tasks with visible solutions. With advances in the mechanisms that support working memory, networks became increasingly able to attend to relevant information in the environment. These developments led to improvements in performance on tasks with visible solutions, paralleling the developmental progression observed in infants. The simulations demonstrate how mechanisms of working memory can subserve perseveration and success on tasks with and without obvious memory demands. In both types of tasks, controlled processing occurs through the activation of task-relevant representations, which provide top-down biasing of other processing pathways. More generally, the simulations demonstrate how common mechanisms can support working memory and attention. PMID- 15829083 TI - Dorsal and ventral stream interaction: contributions from optic ataxia. AB - In monkeys and humans, two functionally specialized cortical streams of visual processing emanating from V1 have been proposed: a dorsal, action-related system and a ventral, perception-related pathway. Traditionally, a separate organization of the two streams is assumed; the extent of functional interaction is unknown. After lesions of the dorsal stream in patients with optic ataxia, it has recently been shown that the ventral perception-related system might contribute to visuomotor processing if movements rely on remembered target positions. The ventral pathway thus seemed to participate in goal-directed movements, a function that previously has been assigned exclusively to the dorsal stream. We wondered whether different types of pointing movements are controlled by switching between two separated cortical pathways or whether a variable interaction of interconnected systems should be assumed. Our study investigated two acute stroke patients with optic ataxia following lesions of the dorsal stream in a delayed pointing task. The delays ranged from 0 to 10 sec. The patients' pointing error decreased in a linear manner with the length of time. The finding suggests a gradual change between dorsal and ventral control of reaching behavior, rather than a sudden switch between two separated cortical processing streams. Although our observations with two patients require further validation, the results suggest that the ventral and dorsal systems interact closely in the sensorimotor control of reaching behavior. PMID- 15829084 TI - The intraparietal sulcus and perceptual organization. AB - The structuring of the sensory scene (perceptual organization) profoundly affects what we perceive, and is of increasing clinical interest. In both vision and audition, many cues have been identified that influence perceptual organization, but only a little is known about its neural basis. Previous studies have suggested that auditory cortex may play a role in auditory perceptual organization (also called auditory stream segregation). However, these studies were limited in that they just examined auditory cortex and that the stimuli they used to generate different organizations had different physical characteristics, which per se may have led to the differences in neural response. In the current study, functional magnetic resonance imaging was used to test for an effect of perceptual organization across the whole brain. To avoid confounding physical changes to the stimuli with differences in perceptual organization, we exploited an ambiguous auditory figure that is sometimes perceived as a single auditory stream and sometimes as two streams. We found that regions in the intraparietal sulcus (IPS ) showed greater activity when 2 streams were perceived rather than 1. The specific involvement of this region in perceptual organization is exciting, as there is a growing literature that suggests a role for the IPS in binding in vision, touch, and cross-modally. This evidence is discussed, and a general role proposed for regions of the IPS in structuring sensory input. PMID- 15829085 TI - Neural correlates of memory for items and for associations: an event-related functional magnetic resonance imaging study. AB - Although results from cognitive psychology, neuropsychology, and behavioral neuroscience clearly suggest that item and associative information in memory rely on partly different brain regions, little is known concerning the differences and similarities that exist between these two types of information as a function of memory stage (i.e., encoding and retrieval). We used event-related functional magnetic resonance imaging to assess neural correlates of item and associative encoding and retrieval of simple images in 18 healthy subjects. During encoding, subjects memorized items and pairs. During retrieval, subjects made item recognition judgments (old vs. new) and associative recognition judgments (intact vs. rearranged). Relative to baseline, item and associative trials activated bilateral medial temporal and prefrontal regions during both encoding and retrieval. Direct contrasts were then performed between item and associative trials for each memory stage. During en- coding, greater prefrontal, hippocampal, and parietal activation was observed for associations, but no significant activation was observed for items at the selected threshold. During recognition, greater activation was observed for associative trials in the left dorsolateral prefrontal cortex and superior parietal lobules bilaterally, whereas item recognition trials showed greater activation of bilateral frontal regions, bilateral anterior medial temporal areas, and the right temporo-parietal junction. Post hoc analyses suggested that the anterior medial temporal activation observed during item recognition was driven mainly by new items, confirming a role for this structure in novelty detection. These results suggest that although some structures such as the medial temporal and prefrontal cortex play a general role in memory, the pattern of activation in these regions can be modulated by the type of information (items or associations) interacting with memory stages. PMID- 15829086 TI - How position, velocity, and temporal information combine in the prospective control of catching: data and model. AB - The cerebral cortex contains circuitry for continuously computing properties of the environment and one's body, as well as relations among those properties. The success of complex perceptuomotor performances requires integrated, simultaneous use of such relational information. Ball catching is a good example as it involves reaching and grasping of visually pursued objects that move relative to the catcher. Although integrated neural control of catching has received sparse attention in the neuroscience literature, behavioral observations have led to the identification of control principles that may be embodied in the involved neural circuits. Here, we report a catching experiment that refines those principles via a novel manipulation. Visual field motion was used to perturb velocity information about balls traveling on various trajectories relative to a seated catcher, with various initial hand positions. The experiment produced evidence for a continuous, prospective catching strategy, in which hand movements are planned based on gaze-centered ball velocity and ball position information. Such a strategy was implemented in a new neural model, which suggests how position, velocity, and temporal information streams combine to shape catching movements. The model accurately reproduces the main and interaction effects found in the behavioral experiment and provides an interpretation of recently observed target motion-related activity in the motor cortex during interceptive reaching by monkeys. It functionally interprets a broad range of neurobiological and behavioral data, and thus contributes to a unified theory of the neural control of reaching to stationary and moving targets. PMID- 15829087 TI - Functional connectivity: integrating behavioral, diffusion tensor imaging, and functional magnetic resonance imaging data sets. AB - In the present study, we combined 2 types of magnetic resonance technology to explore individual differences on a task that required the recognition of objects presented from unusual viewpoints. This task was chosen based on previous work that has established the necessity of information transfer from the right parietal cortex to the left inferior cortex for its successful completion. We used reaction times (RTs) to localize regions of cortical activity in the superior parietal and inferior frontal regions (blood oxygen level-dependent [BOLD] response) that were more active with longer response times. These regions were then sampled, and their signal change used to predict individual differences in structural integrity of white matter in the corpus callosum (using diffusion tensor imaging). Results show that shorter RTs (and associated increases in BOLD response) are associated with increased organization in the splenium of the corpus callosum, whereas longer RTs are associated with increased organization in the genu. PMID- 15829088 TI - Visually induced auditory expectancy in music reading: a behavioral and electrophysiological study. AB - The general aim of this experiment was to investigate the processes involved in reading musical notation and to study the relationship between written music and its auditory representation. It was of main interest to determine whether musicians are able to develop expectancies for specific tonal or atonal auditory events based on visual score alone. Can musicians expect an "atonal" event or will it always sound odd? Moreover, it was of interest to determine whether the modulations in amplitude of a late positive component (P600) described in previous studies are linked to a general mismatch detection process or to specific musical expectancies. Results showed clearly that musicians are able to expect tonal auditory endings based on visual information and are also able to do so for atonal endings, although to a smaller extent. Strong interactions seem to exist between visual and auditory musical codes and visual information seems to influence auditory processing as early as 100 msec. These results are directly relevant for the question of whether music reading is actually music perception. PMID- 15829089 TI - Quantifying stimulus discriminability: a comparison of information theory and ideal observer analysis. AB - Performance in sensory discrimination tasks is commonly quantified using either information theory or ideal observer analysis. These two quantitative frameworks are often assumed to be equivalent. For example, higher mutual information is said to correspond to improved performance of an ideal observer in a stimulus estimation task. To the contrary, drawing on and extending previous results, we show that five information-theoretic quantities (entropy, response-conditional entropy, specific information, equivocation, and mutual information) violate this assumption. More positively, we show how these information measures can be used to calculate upper and lower bounds on ideal observer performance, and vice versa. The results show that the mathematical resources of ideal observer analysis are preferable to information theory for evaluating performance in a stimulus discrimination task. We also discuss the applicability of information theory to questions that ideal observer analysis cannot address. PMID- 15829090 TI - Nonlinear complex-valued extensions of Hebbian learning: an essay. AB - The Hebbian paradigm is perhaps the best-known unsupervised learning theory in connectionism. It has inspired wide research activity in the artificial neural network field because it embodies some interesting properties such as locality and the capability of being applicable to the basic weight-and-sum structure of neuron models. The plain Hebbian principle, however, also presents some inherent theoretical limitations that make it impractical in most cases. Therefore, modifications of the basic Hebbian learning paradigm have been proposed over the past 20 years in order to design profitable signal and data processing algorithms. Such modifications led to the principal component analysis type class of learning rules along with their nonlinear extensions. The aim of this review is primarily to present part of the existing fragmented material in the field of principal component learning within a unified view and contextually to motivate and present extensions of previous works on Hebbian learning to complex-weighted linear neural networks. This work benefits from previous studies on linear signal decomposition by artificial neural networks, nonquadratic component optimization and reconstruction error definition, neural parameters adaptation by constrained optimization of learning criteria of complex-valued arguments, and orthonormality expression via the insertion of topological elements in the networks or by modifying the network learning criterion. In particular, the learning principles considered here and their analysis concern complex-valued principal/minor component/subspace linear/nonlinear rules for complex-weighted neural structures, both feedforward and laterally connected. PMID- 15829091 TI - Difficulty of singularity in population coding. AB - Fisher information has been used to analyze the accuracy of neural population coding. This works well when the Fisher information does not degenerate, but when two stimuli are presented to a population of neurons, a singular structure emerges by their mutual interactions. In this case, the Fisher information matrix degenerates, and the regularity condition ensuring the Cramer-Rao paradigm of statistics is violated. An animal shows pathological behavior in such a situation. We present a novel method of statistical analysis to understand information in population coding in which algebraic singularity plays a major role. The method elucidates the nature of the pathological case by calculating the Fisher information. We then suggest that synchronous firing can resolve singularity and show a method of analyzing the binding problem in terms of the Fisher information. Our method integrates a variety of disciplines in population coding, such as nonregular statistics, Bayesian statistics, singularity in algebraic geometry, and synchronous firing, under the theme of Fisher information. PMID- 15829092 TI - Neurons tune to the earliest spikes through STDP. AB - Spike timing-dependent plasticity (STDP) is a learning rule that modifies the strength of a neuron's synapses as a function of the precise temporal relations between input and output spikes. In many brains areas, temporal aspects of spike trains have been found to be highly reproducible. How will STDP affect a neuron's behavior when it is repeatedly presented with the same input spike pattern? We show in this theoretical study that repeated inputs systematically lead to a shaping of the neuron's selectivity, emphasizing its very first input spikes, while steadily decreasing the postsynaptic response latency. This was obtained under various conditions of background noise, and even under conditions where spiking latencies and firing rates, or synchrony, provided conflicting informations. The key role of first spikes demonstrated here provides further support for models using a single wave of spikes to implement rapid neural processing. PMID- 15829093 TI - Rate and synchrony in feedforward networks of coincidence detectors: analytical solution. AB - We provide an analytical recurrent solution for the firing rates and cross correlations of feedforward networks with arbitrary connectivity, excitatory or inhibitory, in response to steady-state spiking input to all neurons in the first network layer. Connections can go between any two layers as long as no loops are produced. Mean firing rates and pairwise cross-correlations of all input neurons can be chosen individually. We apply this method to study the propagation of rate and synchrony information through sample networks to address the current debate regarding the efficacy of rate codes versus temporal codes. Our results from applying the network solution to several examples support the following conclusions: (1) differential propagation efficacy of rate and synchrony to higher layers of a feedforward network is dependent on both network and input parameters, and (2) previous modeling and simulation studies exclusively supporting either rate or temporal coding must be reconsidered within the limited range of network and input parameters used. Our exact, analytical solution for feedforward networks of coincidence detectors should prove useful for further elucidating the efficacy and differential roles of rate and temporal codes in terms of different network and input parameter ranges. PMID- 15829094 TI - Independent variable time-step integration of individual neurons for network simulations. AB - Realistic neural networks involve the coexistence of stiff, coupled, continuous differential equations arising from the integrations of individual neurons, with the discrete events with delays used for modeling synaptic connections. We present here an integration method, the local variable time-step method (lvardt), that uses separate variable-step integrators for individual neurons in the network. Cells that are undergoing excitation tend to have small time steps, and cells that are at rest with little synaptic input tend to have large time steps. A synaptic input to a cell causes reinitialization of only that cell's integrator without affecting the integration of other cells. We illustrated the use of lvardt on three models: a worst-case synchronizing mutual-inhibition model, a best-case synfire chain model, and a more realistic thalamocortical network model. PMID- 15829095 TI - Synaptic shot noise and conductance fluctuations affect the membrane voltage with equal significance. AB - The subthreshold membrane voltage of a neuron in active cortical tissue is a fluctuating quantity with a distribution that reflects the firing statistics of the presynaptic population. It was recently found that conductance-based synaptic drive can lead to distributions with a significant skew. Here it is demonstrated that the underlying shot noise caused by Poissonian spike arrival also skews the membrane distribution, but in the opposite sense. Using a perturbative method, we analyze the effects of shot noise on the distribution of synaptic conductances and calculate the consequent voltage distribution. To first order in the perturbation theory, the voltage distribution is a gaussian modulated by a prefactor that captures the skew. The gaussian component is identical to distributions derived using current-based models with an effective membrane time constant. The well-known effective-time-constant approximation can therefore be identified as the leading-order solution to the full conductance-based model. The higher-order modulatory prefactor containing the skew comprises terms due to both shot noise and conductance fluctuations. The diffusion approximation misses these shot-noise effects implying that analytical approaches such as the Fokker-Planck equation or simulation with filtered white noise cannot be used to improve on the gaussian approximation. It is further demonstrated that quantities used for fitting theory to experiment, such as the voltage mean and variance, are robust against these non-Gaussian effects. The effective-time-constant approximation is therefore relevant to experiment and provides a simple analytic base on which other pertinent biological details may be added. PMID- 15829097 TI - A subjective distance between stimuli: quantifying the metric structure of representations. AB - As subjects perceive the sensory world, different stimuli elicit a number of neural representations. Here, a subjective distance between stimuli is defined, measuring the degree of similarity between the underlying representations. As an example, the subjective distance between different locations in space is calculated from the activity of rodent's hippocampal place cells and lateral septal cells. Such a distance is compared to the real distance between locations. As the number of sampled neurons increases, the subjective distance shows a tendency to resemble the metrics of real space. PMID- 15829098 TI - Image segmentation by networks of spiking neurons. AB - A network of leaky integrate-and-fire (IAF) neurons is proposed to segment gray scale images. The network architecture with local competition between neurons that encode segment assignments of image blocks is motivated by a histogram clustering approach to image segmentation. Lateral excitatory connections between neighboring image sites yield a local smoothing of segments. The mean firing rate of class membership neurons encodes the image segmentation. A weight modification scheme is proposed that estimates segment-specific prototypical histograms. The robustness properties of the network implementation make it amenable to an analog VLSI realization. Results on synthetic and real-world images demonstrate the effectiveness of the architecture. PMID- 15829099 TI - Neural modeling of an internal clock. AB - We studied a simple random recurrent inhibitory network. Despite its simplicity, the dynamics was so rich that activity patterns of neurons evolved with time without recurrence due to random recurrent connections among neurons. The sequence of activity patterns was generated by the trigger of an external signal, and the generation was stable against noise. Moreover, the same sequence was reproducible using a strong transient signal, that is, the sequence generation could be reset. Therefore, a time passage from the trigger of an external signal could be represented by the sequence of activity patterns, suggesting that this model could work as an internal clock. The model could generate different sequences of activity patterns by providing different external signals; thus, spatiotemporal information could be represented by this model. Moreover, it was possible to speed up and slow down the sequence generation. PMID- 15829100 TI - Mirror symmetric topographic maps can arise from activity-dependent synaptic changes. AB - Multiple adjacent, roughly mirror-image topographic maps are commonly observed in the sensory neocortex of many species. The cortical regions occupied by these maps are generally believed to be determined initially by genetically controlled chemical markers during development, with thalamocortical afferent activity subsequently exerting a progressively increasing influence over time. Here we use a computational model to show that adjacent topographic maps with mirror-image symmetry can arise from activity-dependent synaptic changes whenever the distribution radius of afferents sufficiently exceeds that of horizontal intracortical interactions. Which map edges become adjacent is strongly influenced by the probability distribution of input stimuli during map formation. Our results suggest that activity-dependent synaptic changes may play a role in influencing how adjacent maps become oriented following the initial establishment of cortical areas via genetically determined chemical markers. Further, the model unexpectedly predicts the occasional occurrence of adjacent maps with a different rotational symmetry. We speculate that such atypically oriented maps, in the context of otherwise normally interconnected cortical regions, might contribute to abnormal cortical information processing in some neurodevelopmental disorders. PMID- 15829101 TI - Stochastic optimal control and estimation methods adapted to the noise characteristics of the sensorimotor system. AB - Optimality principles of biological movement are conceptually appealing and straightforward to formulate. Testing them empirically, however, requires the solution to stochastic optimal control and estimation problems for reasonably realistic models of the motor task and the sensorimotor periphery. Recent studies have highlighted the importance of incorporating biologically plausible noise into such models. Here we extend the linear-quadratic-gaussian framework- currently the only framework where such problems can be solved efficiently--to include control-dependent, state-dependent, and internal noise. Under this extended noise model, we derive a coordinate-descent algorithm guaranteed to converge to a feedback control law and a nonadaptive linear estimator optimal with respect to each other. Numerical simulations indicate that convergence is exponential, local minima do not exist, and the restriction to nonadaptive linear estimators has negligible effects in the control problems of interest. The application of the algorithm is illustrated in the context of reaching movements. A Matlab implementation is available at www.cogsci.ucsd.edu/~todorov. PMID- 15829105 TI - Cyclosporine A induced changes to plasma and erythrocyte antioxidant defences. AB - Organ transplant recipients develop pronounced cardiovascular disease, and decreased antioxidant capacity in plasma and erythrocytes is associated with the pathogenesis of this disease. These experiments tested the hypothesis that the immunosuppressant cyclosporine A (CsA) alters erythrocyte redox balance and reduces plasma antioxidant capacity. Female Sprague-Dawley rats were randomly assigned to a control or CsA treated group. Treatment animals received 25 mg/kg/day of CsA via intraperitoneal injection for 18 days. Control rats were injected with the same volume of the vehicle. Three hours after the final CsA injection, rats were exsanguinated and plasma analysed for total antioxidant status (TAS), alpha-tocopherol, malondialdehyde (MDA), and creatinine. Erythrocytes were analysed for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glucose-6-phosphate dehydrogenase (G6PD) activities, alpha-tocopherol, and MDA. CsA administration resulted in a significant (P < 0.05) decrease in plasma TAS and significant increases (P < 0.05) in plasma creatinine and MDA. Erythrocyte CAT was significantly (P < 0.05) increased in CsA treated rats compared to controls. There were no significant differences (P > 0.05) in erythrocyte SOD, GPX, G6PD, alpha-tocopherol or MDA between groups. In summary, CsA alters erythrocyte antioxidant defence and decreases plasma total antioxidant capacity. PMID- 15829106 TI - Antioxidative properties of natural polyamines and dimethylsilane analogues. AB - Structural analogues of natural polyamines, which contain a -Si(CH3)2 group in the central carbon chain, have previously been found to be cytotoxic to various tumor cell lines in vitro and to inhibit tumor cell growth in experimentally grafted animals. In the present study, the antioxidative properties of dimethylsilane polyamine analogues were analyzed in comparison with the natural polyamines. Reactivities of these various polyamines against superoxide anions (generated from the hypoxanthine/xanthine oxidase reaction) and peroxyl radicals (produced from the thermal decomposition of water-soluble 2,2'-azo-bis-[2 amidinopropane] hydrochloride) were investigated. The dimethysilane analogues, and more particularly the hexamine derivative, exhibited the highest scavenging efficiency towards these two reactive oxygen species (ROS). Furthermore, analysis of their ability to prevent hydroxyl radical formation and to trap this ROS showed that the efficiency of the hexamine as a metal chelator and hydroxyl radical scavenger is similar to that of spermine. The higher antioxidant efficiency of the dimethylsilane polyamine analogues with respect to spermidine, together with their ability to displace this polyamine, essential for the promotion of cell growth, from its cellular anionic binding sites that are particularly prone to oxidation, could be biologically relevant and contribute to their in vivo cytotoxic effect and anti-tumor activity. Further experiments will be necessary to demonstrate clearly the relationship between their antioxidant properties and their antiproliferative effects. PMID- 15829107 TI - Biting reduces acute stress-induced oxidative stress in the rat hypothalamus. AB - We investigated the inhibitory effect of para-masticatory activity, namely biting, on restraint stress-induced oxidative stress. A blood brain barrier permeable nitroxyl spin probe, 3-methoxycarbonyl-2,2,5,5,-tetramethylpyrrolidine 1-oxyl (MC-PROXYL), was administered to rats and L-band electron spin resonance (ESR) and ESR-computerized tomography (ESR-CT) imaging were used to show that the decay rate constant of MC-PROXYL in the hypothalamus of isolated brain after 30 min of restraint stress was more rapid than in unrestrained control rats, suggesting that restraint was associated with oxidative stress. Interestingly, biting during restraint stress caused the decay rate constant of MC-PROXYL in isolated brain to approach that of the control group. These observations suggest that biting suppresses oxidative stress induced by restraint stress, and that the anti-stress effect of masticatory motor activity movements, such as biting, are important for reducing the adverse effects associated with exposure to psychological stressors. PMID- 15829108 TI - In vitro effect of hydrostatic pressure exposure on hydroxyl radical production in fish red muscle. AB - The effects of hydrostatic pressure on reactive oxygen species (ROS) production have been studied in vitro on fish red muscle fibres. In the eel, Anguilla anguilla, previous studies have shown that hydrostatic pressure acclimatization improves oxidative phosphorylation efficiency together with a supposed concomitant decrease in electron leak and ROS production. In order to test the hypothesis of an electron leak decrease under pressure, hydroxyl radical (HO*) production and oxygen consumption were measured on fish red muscle fibres directly exposed to hydrostatic pressure. Experiments were performed under two conditions--atmospheric pressure and hydrostatic pressure (16.1 MPa)--on eel and trout (which exhibit low- and high-pressure sensitivity, respectively). This work has permitted, first, the validation of an indirect HO* measurement (in vitro) on fish red muscle and the documentation of reference values for fish. Second, at atmospheric pressure, results show higher oxygen consumption for trout (+40%) than for eel which is accompanied by higher HO* production (+90%); in addition, both species present a positive relationship between HO* production and oxygen consumption. Hydrostatic pressure exposure reverses this relationship for eel but not for trout. These preliminary results only partially verify the proposed hypothesis and further experiments are needed. PMID- 15829109 TI - H2O2 directly activates inositol 1,4,5-trisphosphate receptors in endothelial cells. AB - The mechanisms of H2O2-induced Ca2+ release from intracellular stores were investigated in human umbilical vein endothelial cells. It was found that U73122, the selective inhibitor of phospholipase C, could not inhibit the H2O2-induced cytosolic Ca2+ mobilization. No elevation of inositol 1,4,5-trisphosphate (IP3) was detected in cells exposed to H2O2. By loading mag-Fura-2, a Ca2+ indicator, into intracellular store, the H2O2-induced Ca2+ release from intracellular calcium store was directly observed in the permeabilized cells in a dose dependent manner. This release can be completely blocked by heparin, a well-known antagonist of IP3 receptor, indicating a direct activation of IP3 receptor on endoplasmic reticulum (ER) membrane by H2O2. It was also found that H2O2 could still induce a relatively small Ca2+ release from internal stores after the Ca2+ ATPase on ER membrane and the Ca2+ uptake to mitochondria were simultaneously inhibited by thapsigargin and carbonyl cyanide p-trifluoromethoxyphenyl hydrazone. The later observation suggests that a thapsigargin-insensitive non mitochondrial intracellular Ca2+ store might be also involved in H2O2-induced Ca2+ mobilization. PMID- 15829110 TI - Electron paramagnetic resonance study of the generation of reactive oxygen species catalysed by transition metals and quinoid redox cycling by inhalable ambient particulate matter. AB - A range of epidemiological studies in the 1990s showed that exposure to ambient particulate matter (PM) is associated with adverse health effects in the respiratory system and increased morbidity and mortality rates. Oxidative stress has emerged as a pivotal mechanism that underlies the toxic pulmonary effects of PM. A key question from a variety of studies was whether the adverse health effects of PM are mediated by the carbonaceous particles of their reactive chemical compounds adsorbed into the particles. Experimental evidence showed that PM contains redox-active transition metals, redox cycling quinoids and polycyclic aromatic hydrocarbons (PAHs) which act synergistically to produce reactive oxygen species (ROS). Fine PM has the ability to penetrate deep into the respiratory tree where it overcomes the antioxidant defences in the fluid lining of the lungs by the oxidative action of ROS. From a previous study [Valavanidis A, Salika A, Theodoropoulou A. Generation of hydroxyl radicals by urban suspended particulate air matter. The role of iron ions. Atmospher Environ 2000; 34 : 2379-2386], we established that ferrous ions in PM play an important role in the generation of hydroxyl radicals in the presence of hydrogen peroxide (H2O2). In the present study, we investigated the synergistic effect of transition metals and persistent quinoid and semiquinone radicals for the generation of ROS without the presence of H2O2. We experimented with airborne particulate matter, such as TSPs (total suspended particulates), fresh automobile exhaust particles (diesel, DEP and gasoline, GEP) and fresh wood smoke soot. Using electron paramagnetic resonance (EPR), we examined the quantities of persistent free radicals, characteristic of a mixture of quinoid radicals with different structures and a carbonaceous core of carbon-centred radicals. We extracted, separated and analysed the quinoid compounds by EPR at alkaline solution (pH 9.5) and by TLC. Also, we studied the direct production of superoxide anion and the damaging hydroxyl radical in aqueous and in DMSO suspensions of PM without H2O2. From these results, it is suggested that the cytotoxic and carcinogenic potential of PM can be partly the result of redox cycling of persistent quinoid radicals, which generate large amounts of ROS. In the second phase, the water-soluble fraction of PM elicits DNA damage via reactive transition metal-dependent formation of hydroxyl radicals, implicating an important role for hydrogen peroxide. Together, these data indicate the importance of mechanisms involving redox cycling of quinones and Fenton-type reactions by transition metals in the generation of ROS. These results are supported by recent studies indicating cytotoxic effects, especially mitochondrial damage, by PM extracts and differential mechanisms of cell killing by redox cycling quinones. PMID- 15829111 TI - Dietary supplementation with (R)-alpha-lipoic acid reverses the age-related accumulation of iron and depletion of antioxidants in the rat cerebral cortex. AB - Accumulation of divalent metal ions (e.g. iron and copper) has been proposed to contribute to heightened oxidative stress evident in aging and neurodegenerative disorders. To understand the extent of iron accumulation and its effect on antioxidant status, we monitored iron content in the cerebral cortex of F344 rats by inductively coupled plasma atomic emission spectrometry (ICP-AES) and found that the cerebral iron levels in 24-28-month-old rats were increased by 80% (p<0.01) relative to 3-month-old rats. Iron accumulation correlated with a decline in glutathione (GSH) and the GSH/GSSG ratio, indicating that iron accumulation altered antioxidant capacity and thiol redox state in aged animals. Because (R)-alpha-Lipoic acid (LA) is a potent chelator of divalent metal ions in vitro and also regenerates other antioxidants, we monitored whether feeding LA (0.2% [w/w]; 2 weeks) could lower cortical iron and improve antioxidant status. Results show that cerebral iron levels in old LA-fed animals were lower when compared to controls and were similar to levels seen in young rats. Antioxidant status and thiol redox state also improved markedly in old LA-fed rats versus controls. These results thus show that LA supplementation may be a means to modulate the age-related accumulation of cortical iron content, thereby lowering oxidative stress associated with aging. PMID- 15829116 TI - Determination of itraconazole in human plasma by high-performance liquid chromatography with solid-phase extraction. AB - BACKGROUND: A practical, simple, high-performance liquid chromatographic (HPLC) method is needed for the determination of itraconazole in clinical plasma samples. METHODS: Itraconazole and bifonazole (internal standard) were extracted from plasma using a C8-bonded solid-phase cartridge, separated by C8 reversed phase HPLC, and quantified by ultraviolet absorption at 263 nm. RESULTS: This new method enabled the determination of itraconazole in the concentration range of 10.0-500.0 microg/L. The detection limit of itraconazole was 5.0 microg/L. The mean recovery of itraconazole added to plasma was more than 89.1%, with a coefficient of variation of less than 6.9%. We applied this method for the determination of plasma itraconazole in volunteers treated daily with a 200 mg oral capsule of itraconazole for four days. We monitored the plasma level of itraconazole for the following 24 h and obtained the mean area under the plasma concentration-time curve from 0 to 24 h (AUC(0-24) ) value of 4358.9 +/- 1933.4 microg h/L. CONCLUSION: Our new method will be clinically useful for accurately monitoring the plasma concentration of itraconazole in patients under treatment. PMID- 15829117 TI - Malondialdehyde, glutathione, glutathione peroxidase and homocysteine levels in type 2 diabetic patients with and without microalbuminuria. AB - BACKGROUND: High levels of homocysteine and oxidative stress are known to be associated with premature vascular disease in type 2 diabetes mellitus (DM). The aim of this study was to estimate homocysteine levels and oxidant-antioxidant status and to determine the relationship between them in type 2 diabetic patients with and without microalbuminuria. METHODS: Fasting blood samples were obtained from 48 diabetic patients (17 with and 31 without microalbuminuria) and 20 healthy subjects. Serum total homocysteine (tHcy), plasma malondialdehyde (MDA) erythrocyte glutathione (GSH) and glutathione peroxidase (GPx) activity were measured in these patients and the results were compared with those of controls who were chosen among healthy subjects. RESULTS: MDA levels were found to be significantly lower and GSH levels and GPx activities were found to be significantly higher in control subjects when compared with patients with and without microalbuminuria (MDA: P < 0.0001, P < 0.0001; GSH: P < 0.0001, P < 0.0001; GPx: P < 0.0001, P < 0.001, respectively). MDA levels were found to be significantly higher in patients with microalbuminuria compared with patients without microalbuminuria (P < 0.0001), while similarly GSH levels were found to be significantly lower in patients with microalbuminuria (P < 0.0001). Although there were no significant differences with respect to tHcy levels and GPx activities between the microalbuminuric and normoalbuminuric patients (P > 0.05), there was a significant difference with respect to tHcy levels between healthy controls and patients with microalbuminuria (P < 0.05). The serum levels of tHcy correlated best with plasma MDA and erythrocyte GSH concentrations in all diabetic patients (r = 0.549, P < 0.0001; r = -0.385, P<0.01). CONCLUSION: Decreased antioxidant levels, increased lipid peroxidation and increased tHcy levels were observed in patients with microalbuminuria. These changes may contribute to vascular disease, which is particularly prevalent in type 2 DM patients with microalbuminuria. PMID- 15829118 TI - Prednisolone measurement in human serum using liquid chromatography tandem mass spectrometry. AB - Prednisolone is a commonly prescribed corticosteroid used in the treatment of many diseases. Despite high doses of prednisolone, some patients appear to have subtherapeutic concentrations of the drug. It would be useful to measure prednisolone in this group to determine if they have poor absorption or compliance. Hence, we have developed a liquid chromatography-tandem mass spectrometry method for the determination of prednisolone in serum. Chromatography was performed using a C18 column, giving a retention time for both prednisolone and deuterated prednisolone (internal standard) of 1.6 min. Two transitions were monitored for both prednisolone and deuterated prednisolone. These were m/z 361.2 > 343.0 and m/z 361.2 > 146.9 for prednisolone, and m/z 367.2 > 349.0 and m/z 367.2 > 149.9 for the internal standard. The intra- and inter-batch imprecision was < 7% in both cases over a concentration range of 62.5 750 microg/L. The imprecision at the lower limit was 8%, the lower limit of quantitation was determined to be 30 microg/L and the method was linear up to 5000 microg/L. The method allows rapid prednisolone analysis because of a simplified sample extraction step, and has a cycle time of 3.5 min. PMID- 15829119 TI - Measurement of urinary free cortisol using liquid chromatography-tandem mass spectrometry: comparison with the urine adapted ACS:180 serum cortisol chemiluminescent immunoassay and development of a new reference range. AB - BACKGROUND: The measurement of urinary free cortisol (UFC) is commonly used in the investigation of possible Cushing's syndrome. With the recent availability of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in hospital laboratories, we wanted to develop a specific UFC LC-MS/MS method and compare it with our current immunoassay method and develop a new LC-MS/MS reference range if required. METHODS: A UFC LC-MS/MS method using deuterated cortisol as an internal standard was optimized using solid-phase extraction as a clean-up procedure. The multiple reaction-monitoring transitions used for the detection of cortisol and deuterated cortisol were 363.1 > 121 and 365.1 > 121.8, respectively. The method was investigated regarding precision, linearity, sensitivity, recovery and interference. UFC was measured by the in-house urine adapted ACS:180 serum cortisol immunoassay and the developed LC-MS/MS method in 110 urine samples from patients being investigated for possible Cushing's syndrome. RESULTS: The within batch precisions (n = 25) of the LC-MS/MS method were 7.6%, 4.5% and 3.3% at 25.0 nmol/L, 49.6 nmol/L and 344.6 nmol/L, respectively; the between-batch precisions (n = 10) were 9.4%, 9.4% and 8.4%, respectively, at these concentrations. The method is sensitive down to 5 nmol/L and linear up to at least 1000 nmol/L. The method showed adequate cortisol recovery and no interference from the numerous drugs and steroids tested. The total run time for 20 samples, including sample preparation, was 120 min. A scatter plot of paired UFC measurements on the LC MS/MS and the ACS:180 gave the equation: LC-MS/MS = 0.408 (ACS:180) + 2.65, r2 = 0.6664. The 24-h measured UFC results on 110 samples (25 men and 85 women) were positively skewed. After log transformation the data were less skewed, and following back transformation of the lower 97.5th centile, the upper limit of normal was 165 nmol/24 h. The 95th centile of the untransformed data was 146 nmol/24 h (n = 110, 25 men and 85 women). Separated by sex, the 95th centile was 152 nmol/24 h for men (n = 25) and 141 nmol/24 h for women (n = 85). CONCLUSIONS: We have developed a UFC LC-MS/MS method with a solid-phase extraction clean-up step. The method shows adequate performance and is suitable for routine laboratory use. The mixed sex (n = 110, men = 25, women = 85) reference range was up to 165 nmol/24 h or 146 nmol/24 h, depending on how the data are manipulated. PMID- 15829120 TI - Correction and reporting of potassium results in haemolysed samples. AB - BACKGROUND: Potassium is usually the most important analyte affected by in vitro haemolysis and the result obtained may falsely indicate or disguise a life threatening abnormality and so give rise to inappropriate treatment. The purpose of the study was to provide a solution to the problem of reporting potassium on haemolysed samples, taking into account both clinical needs and analytical concerns (inter-individual and inter-sample variability). METHODS: Using a new procedure that mimics the collection process in an actual clinical setting, haemolysed samples were prepared from 41 volunteers with a range of inter individual factors - haemoglobin 80-173 g/L, red blood cells 2.42-6.77 x 10(12)/L, leucocytes 3.0-306 x 10(9) /L and platelets 31-710 x 10(9)/L - in order to develop a more accurate correction equation using a haemolytic index (HI) corresponding to g Hb/L in plasma. RESULTS: The mean (range) potassium increase was 0.0036 mmol/L (0.0029-0.0053 mmol/L) per unit HI. The following equation was developed to estimate potassium increase per HI, in order to compensate approximately for potassium leakage in haemolysed samples: Corrected K+ = Measured K+ -(HI x 0.004). CONCLUSION: The balanced solution is this: instead of reporting the post-haemolysis corrected potassium result a qualitative comment is given, indicating the likely range of the potassium concentration. If the potassium result is in a critically low or high range, it is communicated promptly to the requesting clinician. PMID- 15829121 TI - Haemoglobin A1c: evaluation of three point of care analysers for use in a paediatric diabetes clinic. AB - BACKGROUND: The measurement of haemoglobin A1c(HbA1c) by high-performance liquid chromatography (HPLC) is generally deemed unsuitable for point of care testing (POCT) due to its complexity and extended turnaround times (TAT). The aim of this project was to evaluate two new HPLC instruments, the Bio-Rad D10 and the Primus PDQ, as POCT instruments compared with Bayer's DCA2000 HbA1c immunoassay analyser in our paediatric diabetes clinic. METHODS: A total of 228 samples were analysed, of which 160 analyses were performed in our paediatric diabetes clinic. HbA1c results were compared by the Passing-Bablok agreement test, the Bland-Altman difference analysis, within- and between-run imprecision, and TAT. RESULTS: : The agreement test and difference analysis showed a correlation of r2 = 0.96 and a mean HbA1c difference of < 0.1% between the three analysers. The PDQ and the D10 achieved the within-run target coefficient of variation (CV) of < 2% at an HbA1c of 7.5%. Between-run imprecision at an HbA1c of 10.8% produced CV of 3.5%, 2.4% and 1.6% for the D10, DCA2000 and PDQ, respectively. TAT studies confirmed that the PDQ was substantially faster than the DCA2000 and D10. CONCLUSIONS: The PDQ had the shortest TAT, afforded random access and exhibited acceptable imprecision, and hence is the preferred instrument for our POCT environment. PMID- 15829122 TI - Effect of the treatment of brucellosis on leukocyte superoxide dismutase activity and plasma nitric oxide level. AB - BACKGROUND: The mechanisms by which brucellae evade intracellular killing by polymorphonuclear leukocytes are incompletely understood. In this study, we evaluated changes of leukocyte superoxide dismutase (SOD) activity and plasma total nitrite as an indicator of nitric oxide (NO) levels during brucellosis therapy. METHODS: Thirty-two patients with acute brucellosis, 27 patients with chronic brucellosis and 30 healthy controls were included in the study. Patients with acute brucellosis were tested for leukocyte SOD activity and plasma total nitrite levels before, during (21st day), and at the end (45th day) of the combined therapy of rifampicin and doxycycline. The same parameters were also investigated in chronic cases and controls. RESULTS: The SOD activities were lower in patients with acute brucellosis before therapy compared with those 21 and 45 days after starting therapy (P < 0.001). In contrast, total nitrite levels did not change significantly (P > 0.05). CONCLUSIONS: In the present study, leukocyte SOD activity was found to be decreased in patients with acute brucellosis. Enzyme activity was increased by treatment, finally reaching the activity of healthy controls. Using an antioxidant agent in addition to classical antimicrobial therapy for acute brucellosis might be a therapeutic approach. PMID- 15829123 TI - Detection and identification of low-concentration M proteins with CZE/IS; a 'dry' analysis. AB - The accuracy of detection and identification of low-concentration M proteins using capillary zone electrophoresis/immunosubtraction (CZE/IS) plots was systematically studied. To this end, 49 CZE plots (protein spectra; session I) and CZE/IS plots (spectra with corresponding immunosubtraction plots; session II) were judged by participants from 12 Dutch laboratories. Plots were based on mixed sera with a low concentration (0.4, 1 or 4 g/L) of one M protein, as well as normal sera, sera with an oligoclonal or polyclonal increase in immunoglobulins, and sera containing free monoclonal light chains. At 0.4 g/L, nine out of 12 (9/12) different M proteins were hard to detect. At 1 g/L, 6/12 M proteins were detected by > 85% of the participants. At 4 g/L, 10/12 M proteins were detected by all participants. Normal spectra were always judged correctly. At 1 g/L and 4 g/L, detected M proteins were identified correctly by 92% of the participants, versus only 63% at 0.4 g/L. The detection accuracy was only slightly (non significantly) improved by the application of IS plots. Our data indicate that CZE/IS is applicable for most M proteins at concentrations > or = 4 g/L. PMID- 15829124 TI - Trace element concentrations in patients on home enteral feeding: two cases of severe copper deficiency. AB - BACKGROUND: Enteral feeding is the fastest growing area of artificial nutrition, with the annual rate of growth being estimated at 20-25% a year. Previous studies have demonstrated trace element deficiencies in patients on long-term home enteral nutrition (HEN). METHODS: The trace elements zinc, selenium, copper and manganese were measured in blood samples from 37 patients on HEN using atomic absorption spectroscopy. RESULTS: Plasma zinc concentrations (range 7.4-14.4 micromol/L) were below the reference range (12.6-22.0 micromol/L) in 30 patients, plasma selenium concentrations (range 0.73-1.76 micromol/L) were below the reference range (0.8-2.0 micromol/L) in only one patient. Whole blood manganese (range 74-309 nmol/L) and plasma manganese (range 13-51 nmol/L) were above both respective reference ranges (73-210 nmol/L and 9-24 nmol/L) in four patients. Two patients showed severely low plasma copper concentrations of 2.4 micromol/L and 2.5 micromol/L, and responded to treatment with extra copper supplementation. CONCLUSION: Although enteral feeds contain adequate concentrations of trace elements, problems with bioavailability may occur and patients receiving long term enteral feeding should be monitored with regard to plasma trace element concentrations. PMID- 15829125 TI - GC/MS identification of toxic pyrrolizidine alkaloids in traditional remedies given to two sets of twins. AB - Neo- and perinatal mortality is high in most of Africa. Twins are at particular risk of perinatal mortality. The contribution of traditional remedies is seldom addressed. The case histories of two sets of twins who were treated with traditional remedies are described. The herbal remedies were analysed using gas chromatography-mass spectrometry. In one family, both siblings survived, albeit with hepatic damage. In the other family, one twin died within 24 h and the second one month after admission with a diagnosis of veno-occlusive liver disease (VOD). In both cases, the presence of the toxic pyrrolizidine alkaloid retrorsine, which is known to lead to VOD, was identified. Health-care workers should be aware of the possibility of traditional medicine administration as a cause of illness, specifically in twin births. PMID- 15829126 TI - A problem with her lead weight. AB - An autistic patient living in a residential home presented with symptoms of weight loss and abdominal pain. On investigation, she was found to have lead poisoning from an unusual source. Her treatment proved difficult due to failure to cooperate, and a 'best-case' protocol was devised. The positive identification of a source of exposure can be difficult in a single case of exposure, but this was eventually achieved by means of stable lead isotope analysis following painstaking detective work. PMID- 15829127 TI - In end-stage renal failure, does infection lead to elevated plasma aluminium and neurotoxicity? Implications for monitoring. AB - The well-described long-term effects of sustained exposure to aluminium in patients with end-stage renal failure (ESRF) are a result of uptake and storage of aluminium, leading to cellular toxicity. A case is presented suggesting that this aluminium may be mobilizable, and indicating the consequence of such release. A patient on haemodialysis (HD) presented acutely with infection, a raised CRP, decreased conscious level, impaired cognition and agitation. Subsequent neurological recovery over six to seven days appeared to follow the return of markedly elevated plasma aluminium concentrations to basal (i.e. from 25.2 micromol/L to 2.5 micromol/L; reference range < 0.5 micromol/L), coupled with a resolution of the infection. The patient was on long-term aludrox therapy 3 g/day, and showed relative resistance to the exogenous hormone erythropoietin, resulting in a refractory anaemia and suggesting aluminium toxicity. A series of HD patients (n = 5) presenting with bacteraemia, not on aludrox, showed no appreciable rise in the plasma aluminium mean of 1.3 micromol/L (SD 0.9; range 0.6-2.0 micromol/L). We suggest that infection can result in release of tissue aluminium, leading to acutely elevated plasma aluminium concentrations and signs of neurotoxicity. The amount of tissue storage and resultant aluminium release seemed to be related to the use of aluminium hydroxide as a phosphate binder. PMID- 15829128 TI - Testosterone replacement-induced hyperprolactinaemia: case report and review of the literature. AB - Half of all men with prolactin (PRL)-producing macroadenomas present with hypogonadism, decreased libido and impotence, and therefore require testosterone replacement. However, very little is known about the effect of testosterone on prolactinomas. We report a case of an 18-year-old obese man who presented with hypogonadism and hyperprolactinaemia and underwent a transphenoidal hypophysectomy after a computer tomography scan showed the presence of a suprasellar macroadenoma. On separate occasions, we documented a rise in PRL when testosterone replacement was started and a fall in PRL when testosterone replacement was stopped (r = 0.6090, P = 0.0095). Furthermore, imaging studies suggested the possibility of tumour re-growth after testosterone therapy. We hypothesize that the exogenous testosterone was aromatized to oestradiol, which stimulated the release of PRL by the anterior pituitary. This was supported by the increase in oestradiol levels after testosterone replacement, although statistical significance was not achieved due to the availability of only a few data points. This case highlights the need to be aware of testosterone replacement-induced hyperprolactinaemia, an under-recognized complication of androgen replacement in this setting. The use of aromatase inhibitors together with testosterone-replacement therapy or the use of non-aromatizable androgens might be indicated in such patients. Taken together, this report and previous studies show that dopamine agonists apparently do not suppress the hyperprolactinaemia induced by testosterone replacement. PMID- 15829129 TI - Effect of a compensated Jaffe creatinine method on the estimation of glomerular filtration rate. PMID- 15829131 TI - The epidemiology of malaria in Bolifamba, a rural community on the eastern slopes of Mount Cameroon: seasonal variation in the parasitological indices of transmission. AB - The prevalences of malarial parasitaemia, fever, splenomegaly and anaemia and the levels of parasitaemia were investigated, through part of one wet season (in 2001) and the following dry season (in 2002), in 2157 subjects in the village of Bolifamba, in south-western Cameroon. Overall, 55.9% of the villagers checked in the wet season but only 49.5% of those examined in the dry season were found smear-positive for malaria (P<0.0001). Rainfall was found to be significantly associated with the mean level of parasitaemia (P=0.001). The prevalences of fever (40.3% v. 19.6%), splenomegaly (37.4% v. 4.0%) and marked splenomegaly (i.e. a Hackett's score of 2 or higher; 25.8% v. 2.4%) were all significantly higher in the wet season than in the dry (P<0.0001 for each). No seasonal difference was observed, however, in the prevalence of anaemia. Parasitaemia, fever, splenomegaly and anaemia were all significantly more common in the young children investigated (i.e. those aged < 5 years) than in the older subjects. When the data were subjected to a multiple logistic regression, age-group, anaemia, fever, and month of examination were all found to be significantly associated with the presence of malarial parasitaemia. The results of this large scale study, the first of its kind in the Buea district of Cameroon, indicate the intense transmission of malarial parasites in rural Bolifamba, with young children at greatest risk. The data collected provide a useful 'base line' for an ongoing study to assess the immune status of the residents of Bolifamba. PMID- 15829132 TI - Household structure and urban services: neglected targets in the control of visceral leishmaniasis. AB - Visceral leishmaniasis (VL) caused by Leishmania chagasi is a growing public health problem in many parts of the New World. Although several studies have focused on the consequences of environmental damage, human migration and land occupation on the incidence of VL, the effects on the disease of the substandard living conditions that often result from the process of urbanization have not been investigated in detail. The present study was based in the Brazilian city of Teresina, where, since 1980, there have been two large outbreaks of VL (one in 1981-1985 and the other in 1993-1996), each involving at least 1000 newly reported cases. The role of household structure and the provision of urban services in the city, as predictors of the occurrence of VL, was studied in a case-control investigation. After controlling for age, crowding, and the background incidence of VL in the area where the subjects lived, the risk of acquiring the disease was found to be significantly higher for those who lived in houses with an inadequate sewage system and those who had no regular rubbish collection. Improving household structure and providing basic urban services might be effective strategies for controlling the spread of VL in urban areas. PMID- 15829133 TI - Pattern of community compliance with spaced, single-dose, mass administrations of diethylcarbamazine or ivermectin, for the elimination of lymphatic filariasis from rural areas of southern India. AB - Current programmes to eliminate lymphatic filariasis (LF) are largely based on annual mass administrations of single doses of antifilarial drugs. The level and pattern of compliance by the target population are important determinants of the success of such mass drug administrations (MDA). Community compliance was therefore investigated during a study in southern India of the effects, on Wuchereria bancrofti microfilaraemia and transmission, of spaced MDA based on diethylcarbamazine (DEC) or ivermectin (IVM). During six rounds of MDA, the frequency of compliance in the target populations, in the five study villages given DEC and the five given IVM, ranged from 55%-77%. Analysis of the relevant cohort data indicated that about 30% of the villagers had complied with treatment during all six rounds, but 3.5% of those in the DEC arm and 4.0% of those in the IVM arm had never complied with treatment. Most of the villagers (>90%) had received treatment at least once, however, and >60% had each received treatment in at least four of the six rounds. Overall, there was a significant negative correlation (r=-0.78; P=0.008) between the size of the village, in terms of the number of villagers, and the mean frequency of compliance over the six rounds of MDA. The pattern of community compliance was found to be 'semi-systematic', laying between random and systematic. In terms of the elimination of LF, a semi systematic pattern of compliance is worse than random compliance but better than systematic. The relevance of the levels and patterns of compliance to LF control or elimination is discussed. PMID- 15829134 TI - Situation analysis in a large urban area of India, prior to launching a programme of mass drug administrations to eliminate lymphatic filariasis. AB - The main strategy now adopted for the elimination of lymphatic filariasis (LF) is based on mass drug administrations (MDA). Annual administration of antifilarial drugs to 65%-80% of the population at risk of the disease is believed to be necessary if LF is to be eliminated, at least as a public-health problem, within a reasonable time-frame. To facilitate the development of drug-delivery strategies that are sufficient to ensure such high treatment coverages in large urban areas, a situation analysis was undertaken in the Indian city of Chennai. The subjects interviewed came from households with high, moderate, low or very low incomes. A lack of information on the prevalence and socio-economic impact of the disease meant that LF was not viewed as a major pubic-health problem in the study area, even though cases of elephantiasis and hydrocele were detected in 2% 8% and 7%-20% of the households investigated. Overall, 40% of the interviewees from very-low-income households and 78% of those from middle-income households knew that (the parasite causing) elephantiasis was transmitted by mosquitoes. Only 4% of the subjects from high-income areas and 1% of those from low-income areas were aware that filarial infection was a major cause of hydrocele. Most of the subjects (>55% of each of the four socio- economic groups considered) felt that they were not at risk of developing elephantiasis. When specifically asked, only 35% of the subjects from high-income households but 84% of those from low income households said that they would be willing to consume tablets of an antifilarial drug (diethylcarbamazine) in MDA to eliminate LF. It is therefore unclear whether high-income households in urban areas should be included in MDA programmes. The interviewees felt that an intensive campaign of information, education, communication and advocacy would be necessary if an effective MDA based programme were to be implemented. Drug distribution through the health services was the most preferred option.Clearly, factors such as a lack of appreciation of the socio-economic impact of LF, a general belief that the risk of elephantiasis is low, doubts about the need to include all sectors of the eligible population in MDA, and a common dependence on private practitioners make successful MDA against LF in urban areas a challenging task. On the positive side, however, an urban population is often covered by a huge network of colleges, private practitioners, non-governmental organizations and residents' associations, and such networks provide new opportunities in the development of effective drug-delivery strategies. PMID- 15829135 TI - Bancroftian filariasis: patterns of vector abundance and transmission in two East African communities with different levels of endemicity. AB - Intensive monitoring of Wuchereria bancrofti vector abundance and transmission intensity was carried out in two communities, one with high-level endemicity for bancroftian filariasis (Masaika, Tanzania) and the other with low-level (Kingwede, Kenya), on the East African coast. Mosquitoes were collected in light traps, from 50 randomly selected households in each community, once weekly for 1 year. They were identified, dissected and checked for parity and filarial larvae. Anopheles gambiae s. l., An. funestus and Culex quinquefasciatus transmitted W. bancrofti in the two communities but the importance of each of these taxa differed between the communities and by season. The overall vector densities and transmission intensities were significantly higher in Masaika than in Kingwede (the annual biting rate by 3.7 times and the annual transmission potential by 14.6 times), primarily because of differences in the available breeding sites for the vectors and in the vectorial capacity of the predominant vector species. A marked seasonal variation in vector abundance and transmission potential contributed to the complex transmission pattern in the communities. Generally, these indices were higher during and shortly after the rainy seasons than at other times of the year. Considerable differences in W. bancrofti transmission were thus observed between communities within a relatively small geographical area (mainly because of environmentally-determined differences in vector habitats), and these were reflected in the marked differences in infection level in the human populations. The variation in vector abundance, vector composition and transmission intensity in the two communities is discussed in respect to its cause, its effects, and its significance to those attempting to control bancroftian filariasis. PMID- 15829136 TI - Cytotaxonomic description of Simulium kaffaense, a new member of the S. damnosum complex (Diptera: Simuliidae) from south-western Ethiopia. AB - Although Ethiopia is one of the countries worst affected by human onchocerciasis, the exact taxonomic identity of the blackflies acting as the main vectors in the endemic areas has never been determined. A cytotaxonomic analysis of Simulium damnosum s.l. collected from three endemic sites in south-western Ethiopia has now revealed the existence of the 'Kisiwani' form (a non-anthropophilic cytoform that is common in East Africa) and a newly recognized species, Simulium kaffaense. Simulium kaffaense sp. nov. is differentiated from other members of the S. damnosum complex by six fixed inversions and dozens of 'new' floating inversions. The rearing of egg batches from some of the biting adult females, to larvae or adults, indicated that the human-biting blackflies were all S. kaffaense. As S. kaffaense is not only highly anthropophilic but also, apparently, the only anthropophilic member of the S. damnosum complex present, it is likely to be the main (if not the only) vector of Onchocerca volvulus in the study area. The presence of inversion 1S-1 and a complex inversion possibly involving 1L-3 indicates that S. kaffaense either belongs or is close to the 'Nile' phylogenetic group of S. damnosum s. l. The karyotype frequencies of the inversions in the collections from the three study sites indicate that at least two forms of S. kaffaense, here designated 'Bebeka' and 'Jimma', were caught. The taxonomy and medical importance of S. kaffaense are discussed. PMID- 15829137 TI - Descriptions of members of the Simulium damnosum complex (Diptera: Simuliidae) from southern Africa, Ethiopia and Tanzania. AB - This paper presents cytotaxonomic details of five populations of the Simulium damnosum complex from South Africa, Swaziland and Ethiopia. The 'Nkusi SW' and 'Pienaars' forms are newly designated members of the complex from South Africa, but the taxonomic rank of an isolate indistinguishable chromosomally from the 'Nkusi' cytoform remains unclear. From Ethiopia two cytoforms were identified, one of which shares two diagnostic chromosome inversions with the cytoform 'Kisiwani' from Tanzania. The second form belongs to S. kaffaense, and is the suspected local vector of Onchocerca volvulus. In addition, a re-analysis of the cytoform 'Kibwezi' from north-eastern Tanzania provided further insights into its population subdivision, and its genetic and morphological characteristics. Cytotaxonomic similarities between 'Kibwezi', S. mengense and S. pandanophilum, along with their biogeography, indicate a relict status of each of these taxa. PMID- 15829138 TI - Geographical characterization of the triatomine infestations in north-central Guatemala. AB - In an entomological study in 2002, the degree of domestic and peridomestic infestation with triatomine bugs and the geographical distribution of such infestations were investigated in north-central Guatemala. The survey team searched for triatomines in houses constructed with mud walls or thatched roofs, in villages suspected of being infested. The level of infestation observed was lower than that seen in the same area and in eastern Guatemala, in a preliminary survey, 3 years earlier. Most of the infestations detected were of Triatoma dimidiata but even this species was found in <7% of the houses investigated. Infestations with Rhodnius prolixus or other potential vectors of Trypanosoma cruzi were much rarer. The generally low levels of infestation make the elimination of R. prolixus and the reduction of the domestic population of Tri. dimidiata feasible in the study area. The southern part of the study area had higher levels of domestic infestation and colonization than the north, and peridomestic infestation was highest in the south-west. Given such geographical variation in the pattern of infestation, it would seem wise to stratify the study region into areas of high, moderate and low-risk of human-triatomine contact, so that appropriate vector-control strategies can be targeted at the worst-affected areas. Regular entomological surveillance, ideally with community participation, is recommended. Analysis of the relationship between the geographical patterns of infestation and factors such as vegetation, altitude and vector migration would be useful. PMID- 15829139 TI - Severe complications and death in cases of Plasmodium falciparum malaria with sickle-cell trait. PMID- 15829140 TI - The quality of sulfadoxine-pyrimethamine prescriptions, counselling and drug dispensing practices, for children in Kenya. PMID- 15829141 TI - Molecular evidence of Rickettsia helvetica and R. monacensis infections in Ixodes ricinus from Hungary. PMID- 15829145 TI - Acute thoracic aortic dissection presenting as sore throat: report of a case. AB - Acute dissection of the aorta can be one of the most dramatic of cardiovascular emergencies. Its symptoms can occur abruptly and progress rapidly. Prompt recognition and appropriate intervention is crucial. However, not all aortic dissections present with classic symptoms of abrupt chest, back, or abdominal pain, and the diagnosis may be missed. Aortic dissection presenting as a sore throat is quite unusual. A 53-year-old man presented with sore throat as the early symptom of an acute thoracic aortic dissection. Unfortunately, the diagnosis was delayed, and the patient died. Given the high morbidity and mortality after delayed recognition or misdiagnosis, aortic dissection should be considered in the differential diagnosis of a patient presenting with sore throat and normal findings of neck and throat, even when there is no classic symptoms. PMID- 15829146 TI - Thoracic aorta dissection associated with aberrant right subclavian artery: treatment with endovascular stent-graft placement. AB - Dissecting aneurysm is the condition produced by separation of the layers of the arterial wall by circulating blood. Although rare, the coexistence of aortic dissection and aberrant right subclavian artery may be catastrophic. In this study we report the endovascular treatment of a patient with thoracic aorta dissection associated with aberrant right subclavian artery. Aortic clamping proximal to the left subclavian artery in a patient with an aberrant right subclavian artery slows or eliminates flow to both vertebral arteries. Endovascular repair eliminates the complications associated with aortic clamping during surgical repair in the presence of an aberrant right subclavian artery; therefore, it should be considered the treatment of choice in this situation. PMID- 15829147 TI - Lupus cerebritis and steroid psychosis in mixed connective tissue disorder. PMID- 15829148 TI - Elihu Yale and the medicine he promoted: the government general hospital and Madras Medical College, India. AB - Much has been written about the philanthropist Elihu Yale and his life in the Americas and England, where he spent his beginnings and end. Less publicized is his life in India, where he spent the majority of his adult life and where he raised his family. A major contribution of Elihu Yale to medicine in India was his promotion of a local hospital in the major Indian trading port city of Madras. This essay briefly describes the history of that hospital and the medical college that grew out of it. PMID- 15829149 TI - Constructing and dismantling frameworks of disease etiology: the rise and fall of sewer gas in America, 1870-1910. AB - For roughly forty years, from 1870 to 1910, Americans recognized and feared gases emanating from sewers, believing that they were responsible for causing an array of diseases. Fears of sewer gas arose from deeper anxieties toward contact with decomposing organic matter and the vapors emitted from such refuse. These anxieties were exacerbated by the construction of sewers across the country during the mid-to-late-nineteenth century, which concentrated waste emanations and connected homes to one another. The result was the birth of sewer gas and the attribution of sickness and death to it, as well as the development of a host of plumbing devices and, especially, bathroom fixtures, to combat sewer gas. The rise of the germ theory, laboratory science, and belief in disease specificity, however, transformed the threat of sewer gas, eventually replacing it (and the larger fear of miasmas) with the threat of germs. The germ theory framework, by 1910, proved more suitable than the sewer gas framework in explaining disease causation; it is this suitability that often shapes the relationship between science and society. PMID- 15829150 TI - On reductionism in biology: pillars, leaps, and the naive behavioral scientist. PMID- 15829151 TI - Decoding of auditory cortex signals with a LAMSTAR neural network. AB - OBJECTIVES: Each neuron has a specific set of stimuli, which it preferentially responds to (the receptive field of the neuron). For implantable cortical prosthetic devices specific points of the cortex (or groups of neurons) have to be stimulated to create perceptions of sensory stimulus with specific attributes (such as frequency, temporal characteristics, etc). Such applications would need real time decoding of signals. Previously mathematical techniques, such as computing the receptive field (using electrophysiology data) and artificial neural networks (Kohonen network or SOM and back propagation network) have been used to decode neural signals. METHODS: A Large Adaptive Memory Storage and Retrieval (LAMSTAR) neural-network-based decoder was designed to decode responses recorded from neurons in the auditory cortex. It was designed to identify the frequency of the tonal stimuli that elicited a particular discharge rate pattern recorded on two channels of a tungsten wire electrode array. RESULTS: The network functioned efficiently as a decoder with 100% accuracy for the small sample of stimulus-response data used. DISCUSSION: The results show that the network is effective in studying the functional organization of the auditory cortex and other sensory systems. Depending on the input sub-word, information about the kind of stimuli that activates particular parts of the sensory cortex can be studied. PMID- 15829152 TI - Diagnostic value of super-selective bilateral cavernous sinus sampling with hypothalamic stimulating hormone loading in patients with ACTH-producing pituitary adenoma. AB - AIMS: Early diagnosis and early treatment by transsphenoidal surgery is desirable for ACTH-producing pituitary microadenoma, but accurate localization of the functional lesion is not always possible before surgery because magnetic resonance (MR) imaging may provide false negative and/or positive findings. The diagnostic value of super-selective bilateral cavernous sinus sampling with the administration of corticotropin-releasing hormone (CRH) was assessed in patients with functioning ACTH-producing pituitary adenoma. METHODS: Fifteen patients with pituitary adenoma (14 with microadenoma) aged from 23 to 74 years (mean 46.7 years) underwent cavernous sinus sampling with or without the CRH loading test and subsequent transsphenoidal surgery in our institute from October 1997 through to November 2002. MR imaging including dynamic scan failed to detect the adenomatous lesion in all patients. To eliminate the bias due to uneven blood flow in the cavernous sinuses and the multi-hormonal response to CRH administration, the ACTH/FSH ratios were evaluated. The inter-cavernous gradient (ICG) was calculated as the higher/lower ACTH venous blood levels in the right and left cavernous sinuses with or without CRH loading. The adjusted ICG was calculated using the ACTH/FSH ratios. The results were compared with the surgical findings. An ICG of 1.4 or greater was considered to indicate the localization of the responsible lesion. RESULTS: Transsphenoidal surgery revealed the functioning lesion on the right in five cases, the left in six, the midline in three and the bilateral lateral wings (double adenoma) in one. Adjusted ICG with CRH loading had a localization accuracy of 93.3% (14/15), which was significantly higher than that of 73.3% (11/15) using ICG without hypothalamic stimulating hormone loading (p = 0.0402). CONCLUSIONS: Super-selective cavernous sinus sampling with hypothalamic stimulating hormone administration can provide accurate localization of the responsible lesion in patients with ACTH-producing pituitary adenoma. PMID- 15829153 TI - Developing a long-term surviving piglet model of neonatal hypoxic-ischemic encephalopathy. AB - OBJECTIVE: This study was designed to develop a piglet model of neonatal hypoxic ischemic encephalopathy, which would allow for serial assessments of long-term neurodevelopmental impairment. METHODS: In 12 newborn piglets, we produced hypoxia by 8% oxygen breathing for 5-91 minutes. We combined ischemia by reversible bilateral common carotid artery occlusion for varying times. Outcome measures were clinical neurological evaluation, magnetic resonance spectroscopy studies and brain histology. RESULTS: Those animals which received intravenous sedation and no mechanical ventilation showed poor tolerance to hypoxia-ischemia and died early in the course of the experiments. The use of inhalation anesthesia during surgical procedures and mechanical ventilation during hypoxia-ischemia was associated with long-term survival. Seven of eight animals that survived > or = 48 hr showed clinical neurological abnormalities, that later resolved. Magnetic resonance spectroscopy measurements did not change significantly following hypoxia-ischemia. None of the animals had histopathological brain lesions. CONCLUSION: When subjected to acute hypoxia-ischemia, piglets were likely to survive only if they were given such supportive measures as anesthesia and mechanical ventilation. Even with hypoxic-ischemic injury sufficient to produce acute signs of neurological dysfunctions, longterm, stable survival with no evident brain histopathological abnormalities was possible. PMID- 15829154 TI - Observing the carotid debris aspirated during carotid stenting: technical note. AB - OBJECTIVES: Elective carotid stent implantation using a distal protection is increasing. In this article we describe an observation method for carotid plaque debris collection during carotid stenting. METHODS: Endovascular stenting for the right internal carotid artery (ICA) stenosis was performed under distal balloon protection. During balloon protection, 30 ml of blood containing debris was aspirated followed by vigorous saline irrigation flushing any residual debris out into the ipsilateral external carotid circulation. Post-operatively, the aspirated blood was filtered and the debris remained on the membrane. The membrane was stained with hematoxylin-eosin (HE) solution and then mounted onto a glass slide for visualization. RESULTS: Microscopic observation of the slide revealed several debris such as atheromatous plaques and intimal strips. HE staining facilitates the characterization of the debris composition. In addition to the histological evaluation, this technique revealed the particle size as well as the quantity of debris. The resulting HE slide is also suitable for permanent storage. In addition to the qualitative and quantitative qualities, this simple technique requires neither specific instrumentation nor equipment. DISCUSSION: Carotid plaque debris aspirated during carotid artery stenting under distal protection can be filtered and visualized on a permanent glass slide. This simple method allows us to better understand quantity, particle size, and composition of debris. PMID- 15829155 TI - Insulin-like growth factor-I decreased etoposide-induced apoptosis in glioma cells by increasing bcl-2 expression and decreasing CPP32 activity. AB - AIMS: In a variety of tumors, the susceptibility of the tumor cells to apoptotic cell death following chemotherapy is a major determinant of therapeutic outcome. Gliomas are resistant to most chemotherapeutic agents, and its mechanism is not known in detail. In an attempt to understand the mechanism of chemo-resistance, we investigated the roles of insulin-like growth factor-I (IGF-I), IGF-I receptors (IGF-IR), and their relationship with the apoptotic response of two glioma cell lines to etoposide, a chemotherapeutic agent for malignant gliomas. METHODS: Two human glioma cell lines, U-87MG and KNS-42, were used. Etoposide induced cell growth inhibition was quantified using a modified MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide), colorimetric assay. Hoechst 33258 staining, DNA fragmentation assay, and western blot were used for the evaluation of apoptosis. ApoAlert caspase assay was used for measuring the activity of caspase-3 (CPP32) and interleukin-1 beta -converting enzyme (ICE) protease. In addition, the effect of IGF-IR antisense was tested in U-87MG and KNS-42 glioma cell lines. RESULTS: Etoposide inhibited the growth of U-87MG and KNS-42 cells in a concentration-dependent manner. Etoposide increased the expression of wild-type p53, activated CPP32 (but not ICE) activity, and induced apoptosis in these cells. IGF-I prevented etoposide-induced apoptosis by increasing the expression of bcl-2 and decreasing the activity of CPP32. IGF-IR antisense enhanced the apoptotic effect of etoposide. CONCLUSIONS: IGF-I decreased etoposide-induced apoptosis in glioma cells by increasing the expression of bcl-2 and decreasing the activity of CPP32. The antisense of IGF-IR increased etoposide-induced apoptosis. The anti-apoptotic effect of IGF-I and IGF IR might be related to the chemo-resistance of glioma to chemotherapeutic agents. PMID- 15829156 TI - The trigemino-cervical reflex in patients with trigeminal neuralgia. AB - OBJECTIVES: To investigate the central trigeminal system in idiopathic trigeminal neuralgia (TN). MATERIALS AND METHODS: Short latency responses can be recorded in sternocleidomastoid (SCM) muscles after stimulation of the trigeminal nerve (trigemino-cervical reflex). This brainstem reflex was investigated in 40 healthy subjects and in 17 patients suffering from idiopathic TN before and after therapy for 2 months with carbamazepin. RESULTS: Before therapy, six patients presented abnormalities of SCM responses on the painful side, six patients with bilateral abnormalities, and five patients with normal responses. A significant variation in the responses after therapy was found only in the patients with unilateral abnormalities: these patients and the patients with normal reflexes before therapy also had a good response to the therapy with significant pain relief. CONCLUSIONS: Our findings suggest that the trigemino-cervical reflex could be useful in the clinical assessment of TN prior to instituting non-surgical treatment. The bilateral location of the abnormalities in some patients seems to point to a centrally located dysfunction; therefore, this study supports the idea that mechanisms in the central nervous system may play an important role in the pathophysiology of trigeminal neuralgic pain. PMID- 15829157 TI - Pathogenesis of some neurological immune ultrastructural and morphometrical observations on rat thymus. AB - Numerous studies on neuro-immuno-modulation indicate that the thymus is involved in many neurological diseases, including experimental allergic encephalomyelitis (EAE). Twenty Lewis rats were induced for EAE. At X, XII, XX and XXX days post inoculation the animals were killed, and the thymus was recovered and harvested. Specimens of thymus were submitted to morphological light microscopy analysis (1% toluidine blue) and ultra-structural analysis (transmission electron microscopy). Significant morphometric data were collected by examining the images quantitatively and by statistically analysing the values. Our results show that the microenvironment of the thymus is severally involved in acute EAE. Thymocytes and reticular epithelial cells show many changes which are closely related to the pathogenesis of EAE. In particular we observed: (1) inside the cell an increase in intra-cytoplasmic vacuoles, and changes in the thickness of the nuclear membrane, mitochondria, rough endoplasmic reticulum, cellular inter-digitations and cellular electron-density; (2) outside the cell an increase in pericellular translucent halo, intercellular spaces, intercellular contacts and apoptotic and necrotic figures. The evidence of a thymic role in MS may suggest the intriguing therapeutic concept of thymectomy in the management of this neurological disease. PMID- 15829158 TI - The intraluminal thread model revisited: rat strain differences in local cerebral blood flow. AB - There are major differences in the outcome of focal cerebral ischemia between rat strains. This study aimed to investigate whether inter-strain differences exist in the local cerebral blood flow, changes produced by intraluminal middle cerebral artery occlusion (MCAO). Fifty-four male Long-Evans, Sprague-Dawley or Wistar rats were subjected to 60 minutes of transient MCAO, carried out with a silicone-coated 4-0 nylon monofilament. Local cerebral blood flow was monitored on the parietal cortex with laser-Doppler flowmetry. The decrease in local cerebral blood flow during the first 20 minutes of occlusion was significantly greater in Long-Evans (74.6 +/- 8.9%) than in Sprague-Dawley (50.7 +/- 26.8%) and Wistar (49.0 +/- 21.9%) rats. Strain differences in local cerebral blood flow reduction were maintained, or increased, throughout the occlusion period. These results show that the intraluminal model of MCAO produces a more pronounced, persistent and stable local cerebral blood flow reduction, with less interanimal variability, in Long-Evans than in Sprague-Dawley or Wistar rats. PMID- 15829159 TI - Congenital intracranial immature teratoma of the lateral ventricle: a case report and review of the literature. AB - OBJECTIVE: Congenital intracranial tumors are very rare and only account for 0.5 1.5% of all childhood brain tumors. The most common type of these tumors present at birth is teratomas, which represent 0.5% of all intracranial tumors. Most teratomas are midline tumors located predominantly in the sellar and pineal regions. In this study, we report a neonatal intracranial immature teratoma at the lateral ventricle because of its rare location. CASE REPORT: A 3-day-old female neonate presented with a history of irritability, vomiting, and recurrent generalized clonic seizures since birth. A head computed tomographic scan and magnetic resonance imaging disclosed a large tumor filling the right lateral ventricle and extending into the ipsilateral posterior fossa. With right parieto occipital craniotomy, large grayish-white lobulated vascular mass was encountered and total removal of tumor was performed. Histological examination revealed the diagnosis of immature teratoma. CONCLUSION: The prognosis of congenital intracranial immature teratoma is usually poor because the lesions are extensive when they are identified. Prenatal ultrasonography is necessary for the prenatal diagnosis. Fetal magnetic resonance imaging should be made for the evaluation of intracranial tumor. If the tumor is detected before the 24 week of gestation, termination of the pregnancy should be considered. PMID- 15829160 TI - Detection of experimentally induced brain tumors in rats using high resolution computed tomography. AB - OBJECTIVE: The feasibility of high resolution CT imaging for evaluating experimentally-induced brain tumors in rats was assessed. METHODS: The gliosarcoma cell line (9 L/lacZ) was inoculated in 34 male Fischer 344 rats and CT studies were performed prior to and at 4, 7, 9, 12 and 14 days post-tumor cell implantation. Brain imaging pre- and post-contrast was performed and correlated with autopsy findings. RESULTS: Tumors were identified by CT in 19 of the 34 animals after contrast administration and their presence was confirmed at autopsy. Tumors were present at autopsy and not identified by CT scanning in eight additional animals and in the remaining seven animals, the CT scan was normal and no tumors were present at autopsy. The sensitivity and specificity of CT scanning with contrast in detecting tumors in this rat model of gliosarcoma was 70 and 100%, respectively. CONCLUSION: The improved CT technology currently available can be used to identify and follow tumor burden in a rat model of gliosarcoma, and be a good tool to utilize in determining treatment outcomes experimentally, especially when MR imaging is not available. PMID- 15829161 TI - Effects of adenosine and adenosine A2A receptor agonist on motor nerve conduction velocity and nerve blood flow in experimental diabetic neuropathy. AB - This study examined the effects of chronic administration of adenosine and CGS 21680 hydrochloride (adenosine A(2A) receptor agonist) on motor nerve conduction velocity (MNCV), nerve blood flow (NBF) and histology of sciatic nerve in animal model of diabetic neuropathy. Adenosinergic agents were administered for 2 weeks after 6 weeks of streptozotocin-induced (50 mg/kg i.p.) diabetes in male Sprague Dawley rats. Significant reduction in sciatic MNCV and NBF were observed after 8 weeks in diabetic animals in comparison with control (non diabetic) rats. Adenosine (10 mg/kg, i.p.) significantly improved sciatic MNCV and NBF in diabetic rats. The protective effect of adenosine on MNCV and NBF was completely reversed by theophylline (50 mg/kg, i.p.), a non-selective adenosine receptor antagonist, suggesting that the adenosine effect was mediated via adenosinergic receptors. CGS 21680 (0.1 mg/kg, i.p.) significantly improved NBF; however, MNCV was not significantly improved in diabetic rats. At a dose of 1 mg/kg, neither MNCV nor NBF was improved by CGS 21680 in diabetic rats. ZM 241385 (adenosine A(2A) receptor antagonist) prevented the effect of CGS 21680 (0.1 mg/kg, i.p.). Histological changes observed in sciatic nerve were partially improved by the adenosinergic agents in diabetic rats. Results of the present study, suggest the potential of adenosinergic agents in the therapy of diabetic neuropathy. PMID- 15829162 TI - Intraventricular albumin: an optional agent in experimental post-traumatic brain edema. AB - HYPOTHESIS: Human albumin may be effective in the treatment of posttraumatic brain edema due to its hyperoncotic features. Therefore, the aim of the experimental study presented in this paper has two points: the first is to evaluate the efficacy of intraventricular hyperoncotic human albumin on post traumatic brain edema and the second is to try to show the appropriate posttraumatic time window for albumin administration. METHOD: Traumatic brain injury and subsequent edema was formed by a model of impact acceleration injury in rats. Human albumin was administered via intraventricular route by using a stereotactic head holder. All animals in each group were decapitated 24 hours after the procedure and the effect of albumin was evaluated by measurement of tissue specific gravity. RESULTS: Tissue specific gravity decreased in edematous tissue (trauma indicator), increased after albumin administration at the 12th (p < 0.001), and both at the 1st and 12th hour of the trauma (edema treatment; p < 0.001). On the other hand, albumin administered at the 12th, and at both the 1st and 12th hours in the rats without trauma has caused the formation of the brain edema. CONCLUSION: We conclude that human albumin is effective in cytotoxic, but not in vasogenic edema and exerts its best anti-edematous effect at the 12th hour of severe head trauma and this study may help future studies that will try to show the effects of albumin with different time modalities after severe head injury. PMID- 15829163 TI - Methylenetetrahydrofolate reductase gene polymorphisms in patients with cerebral hemorrhage. AB - Elevated plasma total homocysteine (HCY) level is a risk factor for coronary heart disease and ischemic stroke. We investigated relationships between polymorphisms in the methylenetetrahydrofolate reductase (MTHFR) gene, and plasma levels of HCY and folate in patients of Mongolian races who suffered from cerebral ischemia (CI, n = 42) or cerebral hemorrhage (CH, n = 20) and in the 24 age-matched controls. The incidences of both homozygous and heterozygous MTHFR gene mutations in CI (26 and 43%) and in CH (25 and 60%) were significantly higher than those in the controls (8 and 25%). Homozygous MTHFR gene mutation was associated with reduced plasma folate levels, but not with increased plasma HCY levels. Among the subjects with homozygous MTHFR gene mutation, plasma folate levels in CH was significantly lower than those in CI and controls. MTHFR gene mutation in CH was found to be as common as that in CI and was associated with reduced plasma folate levels in the both. In homozygous MTHFR gene mutation, the plasma folate level was profoundly reduced in CH as compared with CI and controls, suggesting that subjects with low plasma folate levels have a predisposition to intracerebral bleeding. PMID- 15829164 TI - Effect of melatonin on cerebral vasospasm following experimental subarachnoid hemorrhage. AB - OBJECT: The current study was undertaken to determine whether melatonin therapy reverses vasospasm and prevents apoptosis by inhibiting lipid peroxidation in an experimental subarachnoid hemorrhage (SAH) model. MATERIALS AND METHODS: The rabbits were divided into four groups as follows: Group 1, SAH + melatonin (5 mg/kg/i.p. BID) simultaneously with SAH (n = 6); Group 2, SAH + melatonin (5 mg/kg/i.p. BID) treated 2 hours after SAH (n = 6); Group 3, control group (n = 4); Group 4, SAH only (n = 6). Light microscopic examinations of the basilar arteries were performed to demonstrate the pathophysiological changes of the arterial wall with hematoxylin- eosin. Apoptosis: Immunohistology using the ApopTag Peroxidase In Situ Apoptosis Detection Kit was used to demonstrate apoptosis in a cross section of basilary arteries. Apoptotic index was calculated as the number of the immunoreactive nuclei per total number of endothelial cells, and expressed as a percentage. RESULTS: The results of measurements of diameters of the vessels between groups were significantly different (p = 0.028). While basilar arteries of the SAH only group showed 57% constriction, Groups 1 and 2 were calculated as 33 and 26% constriction, respectively, compared with the control group (p < 0.05). And also Groups 1 and 2 showed significant protection of apoptosis compared with Group 4. The difference between the four groups was tested by Kruskal-Wallis test and the significance between the two groups was tested by Mann- Whitney U-test. CONCLUSION: Melatonin with its strong antioxidant effect can prevent SAH-induced vasospasm and apoptosis of endothelial cells of vessels. PMID- 15829165 TI - Capnography screening for sleep apnea in patients with acute stroke. AB - Sleep apnea syndrome (SAS) is a prominent clinical feature in acute stroke patients. Diagnosis is usually established by polysomnography or cardio respiratory polygraphy (CRP). Both diagnostic procedures produce high costs, are dependent on the access to a specialized sleep laboratory, and are poorly tolerated by patients with acute stroke. In this study we therefore investigated whether capnography may work as a simple screening tool in this context. In addition to conventional CRP, 27 patients with acute stroke were studied with capnography provided by our standard monitoring system. The trend graphs of the end-tidal CO(2) values (EtCO(2)) were used to determine the capnography-based estimate of the apnea-hypopnea index (AHI(CO2)). Index events were scored when the EtCO(2) value dropped for > 50% of the previous baseline value. We found that the AHI(CO2) correlated significantly with the apnea-hypopnea index measured with conventional CRP (AHI(CRP)) (r = 0.94; p < 0.001). An AHI(CO2) > 5 turned out to be highly predictive of an AHI(CRP) > 10. According to our findings, routinely acquired capnography may provide a reliable estimate of the AHI(CRP). The equipment needed for this screening procedure is provided by the monitoring systems of most intensive care units and stroke units where stroke patients are regularly treated during the first days of their illness. Therefore, early diagnosis of SAS in these patients is made substantially easier. PMID- 15829166 TI - In vivo multiparametric monitoring of brain functions under intracranial hypertension following mannitol administration. AB - OBJECTIVE: Over the last 20 years, mannitol has replaced other osmotic diuretics. Its beneficial effects on intracranial pressure (ICP), cerebral perfusion pressure (CPP), cerebral blood flow (CBF) and brain metabolism are widely accepted. In the present study, we tested the effect of mannitol injection on brain hemodynamic, metabolic, ionic and electrical state in rats exposed to intracranial hypertension. METHODS: The parameters monitored simultaneously included ICP, CBF using the laser Doppler flowmetry, mitochondrial NADH redox state by the fluorometric technique, extracellular K(+) and H(+) levels, DC potential, ECoG, blood pressure and calculated CPP. ICP was elevated to 30 mmHg for 30 minutes and mannitol was injected 15 minutes post-ICP elevation. RESULTS: Our results showed that mannitol decreased ICP, and improved the levels of MAP, CPP and CBF. Moreover, mannitol completely prevented mortality following intracranial hypertension in rats. CONCLUSION: It seems that the multiparametric monitoring approach, used in intracranial hypertension models, is an important tool for brain functional state evaluation. PMID- 15829167 TI - Elevation of plasma oxidized LDL in acute stroke patients is associated with ischemic lesions depicted by DWI and predictive of infarct enlargement. AB - Oxidized low-density lipoprotein (OxLDL) plays a major role in atherosclerosis. We undertook the present study to clarify the relationship between plasma OxLDL and the ischemic volume. We used ELISA to determine plasma OxLDL levels, and performed diffusion- and perfusion-weighted MRI (DWI, PWI) to measure the ischemic volume in 44 ischemic stroke patients. Based on the location of the ischemic lesion, they were divided into three groups: Group I (GI, n = 21) had cortical lesions, Group II (GII, n = 17) had lesions in the basal ganglia or brain stem, and Group III (GIII, n = 6) had massive lesions that involved one entire hemisphere. In GI, but not GII and GIII, plasma OxLDL was significantly higher than in 19 age-matched controls (p < 0.01) and was significantly correlated with the initial ischemic volume visualized on DWI (p = 0.01), PWI (p < 0.01), and the DWI-PWI mismatch (p < 0.05). A persistent increase in plasma OxLDL was associated with enlargement of the ischemic lesion in the early phase after the insult. These findings suggest that elevated plasma OxLDL levels are associated with moderate ischemic damage in patients with cortical lesions (GI), but not those with massive hemispheric lesions (GIII), which may be irreversible. In addition, elevated plasma OxLDL may represent a predictor of enlargement of the ischemic lesion. PMID- 15829168 TI - Cheiro-pedal syndrome following pontine infarction. AB - We report the case of a 64-year-old man with sudden onset of numbness in the right hand and foot. Neurological examinations were normal except for hypersthesia, and hyperalgesia of the right hand and foot. Brain MRI demonstrated a high signal intensity on T2-weighted image and a low signal intensity on T1 weighted image in the left tegmetum of the pons. He was diagnosed with pontine infarction presenting with cheiro-pedal syndrome (CPS). Damage in the sensory pathways can cause CPS. Difference in the threshold may explain the specific sensory pattern in this syndrome. Further examination of the relationship between sensory symptoms and localization on MRI is needed to clarify this syndrome. PMID- 15829169 TI - Oxidative stress and metal content in blood and cerebrospinal fluid of amyotrophic lateral sclerosis patients with and without a Cu, Zn-superoxide dismutase mutation. AB - Hydroxyl radical, ascorbate free radical, superoxide dismutase (SOD) activities, Cu,Zn-SOD protein, Mn-SOD protein, 8-hydroxy-2' -deoxyguanosine (8-OHdG) and metals were compared in red blood cells (RBC), plasma and/or cerebrospinal fluid (CSF) between patients with sporadic amyotrophic lateral sclerosis (SALS), familial ALS (FALS) showing the Leu126Ser mutation in the Cu, Zn-SOD gene and controls. In patients with FALS or SALS, concentrations of hydroxyl radical in blood and ascorbate free radical and 8-OHdG in CSF were higher than control group values, while SOD activities in RBC and CSF were lower. In contrast, Cu, Zn-SOD protein concentrations in RBC were low only in FALS patients. Concentrations of Cu in CSF of SALS patients were higher than in controls. Thus, the pathogenesis of increased oxidative stress differs between SALS patients and FALS patients with a mutant Leu126Ser SOD1 gene. PMID- 15829170 TI - Distribution of artificial cerebral microemboli in stroke patients with patent foramen ovale. AB - OBJECTIVES: Patent foramen ovale (PFO) is considered as an important risk factor for cerebrovascular diseases. Nevertheless, the relationship between the distribution of high-intensity transient signals (HITS), resulting from injection of air mixed with saline and detected by transcranial Doppler (TCD), and clinical cerebrovascular syndromes in these patients has not been investigated. METHODS: Using TCD, we screened 40 patients with stroke or transient ischemic attack (TIA), in whom PFO was proven by transesophageal echocariography (TEE). Of these, 30 patients (75%) with artificially produced HITS either in the middle cerebral artery (MCA) or the basilar artery (BA) were included in the analysis. RESULTS: Nineteen patients had a stroke or TIA in the carotid territory and 11 patients in the vertebro-basilar territory. HITS were found in the MCA in all 30 patients and in 21 of the 30 patients in the BA. Of the latter, ten patients were in the carotid group and 11 patients were in the veretebro-basilar group, p = 0.011. CONCLUSION: There is a significant association between the distribution of artificial HITS and the clinical cerebrovascular syndromes. PMID- 15829171 TI - Spinal cord separation: MRI evidence of healing after omentum-collagen reconstruction. AB - INTRODUCTION: Animal experimentation has demonstrated that omental-collagen bridge reconstruction of a transected spinal cord in cats can result in the growth of axons crossing the transection site which resulted in the return of motor and sensory activity. This paper raises the possibility that a comparable spinal cord reconstruction model could be possible for human application. METHODS: Cats had their spinal cord transected at the T-9 level. This led to a gap at the transection site that was filled with semi-liquid collagen, followed by omental transposition onto the underlying collagen bridge, which had subsequently hardened. A comparable technique was used on a patient who had, as reported by magnetic resonance imaging (MRI), a complete spinal cord transection at the T-6 level. RESULTS: Reconstruction of a transected spinal cord in cats using an omental-collagen bridge resulted in axons that grew across the transection site at the rate of 1 mm/day. Several animals developed forelimb and hindlimb locomotion. The patient in this paper had omental-collagen reconstruction of her cord and has clinically progressed to the point where she can ambulate with the use of a walker. The patient had a spinal cord defect of 4 cm, which, with multiple MRI studies, has shown the longitudinal development of a spinal cord connection in the area of the omental-collagen bridge that connects the proximal and distal ends of the transected spinal cord. CONCLUSION: This report suggests that a transected spinal cord has the ability to heal when the spinal cord separation is reconstructed using an omental-collagen bridge. This technique has led to neurological improvement. PMID- 15829172 TI - Adenosine and the brain (a tribute to the contributions of Dr John W. Phillis): prologue. PMID- 15829173 TI - Purinergic reminiscences. PMID- 15829174 TI - From cAMP to adenosine: an illuminating shift of focus. AB - In a remarkable career, straddling five decades, John Phillis pursued with fierce determination and exceptional energy the main goal of his scientific life, to throw light on the chemical agents that control brain function. Starting in Australia, he settled in North America, first in Canada, then in the USA, where his long tenure at Wayne State brought his career to its culmination. PMID- 15829175 TI - Adenosine sleep theory: how I postulated it. PMID- 15829176 TI - Purine level regulation during energy depletion associated with graded excitatory stimulation in brain. AB - OBJECTIVES: The formation and release of adenosine following graded excitatory stimulation of the brain may serve important physiological functions such as sleep regulation, as well as an early resistance mechanism against excitotoxicity. However, adenosine at high levels may reflect merely the results of obstructed energy metabolism. METHODS: We examined the extent to which levels of adenosine and adenylate energy charge are affected in vivo by graded excitatory stimulations of brain using unilateral intrastriatal injections of glutamatergic agents and head-focused high energy microwaving for accurate and precise measures of purines. RESULTS: Our results confirmed that adenosine levels rise when adenylate energy charge decreases and showed that these increases occurred in three distinct phases with the rate of adenosine formation in each phase increasing as tissue adenylate energy charge was further depleted. In addition, we observed that, in most cases, the effects of focal excitatory stimulation on changes in tissue purine levels were restricted spatially within the immediate vicinity of the injection site; however, when strongly depolarizing stimuli were used, changes in purine levels could be observed in adjacent and, occasionally, even in contralateral brain regions. DISCUSSION: These results provide new insight into purine regulation that occurs under physiologically relevant conditions, such as sleep and during the early stages of brain insults that induce excitotoxicity. PMID- 15829177 TI - Adenosine, glutamate and pH: interactions and implications. AB - Adenosine's role in the nervous system is multifaceted. As the core molecule of adenosine triphosphate (ATP), adenosine exists in equilibrium with the adenine nucleotide pool and contributes to cellular energy charge, a quantification of relative amounts of available ATP, ADP, AMP and adenosine. Beyond participating in overall energy balance and thus in maintaining cellular homeostasis, adenosine critically influences dynamic signaling in the nervous system. In particular, adenosine has an effect on and is affected by excitatory synaptic transmission. This report describes the ubiquitous nature of adenosine's influence, outlines specific scenarios of clinical import and highlights emerging knowledge about the regulation of adenosine. PMID- 15829178 TI - Astrocytes and neurons: different roles in regulating adenosine levels. AB - OBJECTIVES: Adenosine is an endogenous nucleoside that signals through G-protein coupled receptors. Extracellular adenosine is required for receptor activation and two pathways have been identified for formation and cellular release of adenosine. The CLASSICAL pathway relies on intracellular formation of adenosine from adenine nucleotides and cellular efflux of adenosine via equilibrative nucleoside transporters (ENTs). The ALTERNATE pathway involves cellular release of adenine nucleotides, hydrolysis via ecto-5'-nucleotidases and extracellular formation of adenosine. METHODS: A rat model of cerebral ischemia and primary cultures of rat forebrain astrocytes and neurons were used. RESULTS: Using a rat model of cerebral ischemia, the ENT1 inhibitor nitrobenzylmercaptopurine ribonucleoside (NBMPR) significantly increased post-ischemic forebrain adenosine levels and significantly decreased hippocampal neuron injury relative to saline treatment. NBMPR-induced increases in adenosine receptor activation were not detected, suggesting that altering the intracellular:extracellular distribution of adenosine can affect ischemic outcome. Using primary cultures of rat forebrain astrocytes and neurons, adenosine release was evoked by ischemic-like conditions. Dipyridamole, an inhibitor of ENTs, was more effective at inhibiting adenosine release from neurons than from astrocytes. In contrast, alpha , beta-methylene ADP, an inhibitor of ecto-5'-nucleotidase, was effective at inhibiting adenosine release from astrocytes, but not from neurons. Thus, during ischemic-like conditions, neurons released adenosine via the CLASSICAL pathway, while astrocytes released adenosine via the ALTERNATE pathway. DISCUSSION: These cell type differences in pathways for adenosine formation during ischemia may allow transport inhibitors to block simultaneously adenosine release from neurons and adenosine uptake into astrocytes. In principle, this could improve neuronal ATP levels without decreasing adenosine receptor activation. PMID- 15829179 TI - Adenosine, neurodegeneration and neuroprotection. AB - Protection against neuronal damage is a major objective of current research in areas such as stroke medicine, Alzheimer's disease and other neurodegenerative conditions. Adenosine receptors are important modulators of cell survival, and thus agents targeting these receptors could be valuable therapeutic agents. Agonists at A(1) receptors and antagonists at A(2A) receptors are known to protect acutely against neuronal damage caused by toxins or ischemia-reperfusion, and these compounds can also protect against the cell damage inflicted by reactive oxygen species. Even endogenous adenosine may be neuroprotective, since its levels rise substantially in association with a period of ischemia reperfusion. Unfortunately, there is growing evidence that the efficacy of adenosine receptor activation can be reduced by the concomitant activation of glutamate receptors responding to N-methyl-D-aspartate (NMDA), probably acting via the release of nitric oxide. Such problems will need to be resolved before adenosine receptor agonists can proceed far as neuroprotective agents. The use of receptor antagonists may prove a more valuable approach. PMID- 15829180 TI - The protective effect of adenosine A2A receptor antagonism in cerebral ischemia. AB - OBJECTIVES: We reviewed our most recent work on the protective effect of adenosine A(2A)antagonism in cerebral ischemia. METHODS: Focal ischemia was produced in rats by introducing a nylon monofilament pre-coated with silicone through the external carotid artery to occlude the right MCA at its origin. RESULTS: A(2A) antagonism was found protective in the model of permanent focal ischemia induced by the monofilament technique. This methodology provides the possibility of evaluating the protection against the outflow of excitatory amino acids and against an acute motor disturbance, i.e.contralateral turning to the ischemic side in the first hours after ischemia in awake rats. Hours later, a definite neurological deficit and necrotic neuronal damage can be evaluated. DISCUSSION: Our results suggest that A(2A) antagonism may be protective from the earliest up to several hours after the ischemic event. PMID- 15829181 TI - Adenosine and the regulation of cerebral blood flow. AB - OBJECTIVES: This review summarizes the 30 year effort of my collaborator and mentor Dr J. W. Phillis to establish the role of adenosine in the regulation of cerebral blood flow. METHODS: While most of the experiments described utilized the rat cerebral cortex as a model, several different and complementary methodologies were employed. Superfusate samples were collected from the cortical surface and analysed for purines using HPLC. Laser-Doppler flowmetry was utilized to measure blood flow in the pial vasculature, while pial diameters were monitored by videomicroscopy. An additional series of experiments looked at coronary blood flow in a Langendorff preparation. RESULTS: Adenosine is released from the cortex in response to decreased nutrient supply (hypoxia/ ischemia) and during conditions that mimic alterations in the extracellular environment associated with increased metabolism. The application of pharmacological agents that alter adenosine metabolism resulted in the appropriate alterations in ECF adenosine levels and also in blood flow. Selective blockade of the adenosine A(2A) receptor reduced the pial vasodilation evoked by hypercapnoea. Results from the isolated rat heart, utilizing similar agents, support a role for adenosine in the regulation of coronary blood flow during respiratory and metabolic acidosis. DISCUSSION: Adenosine is released when there is a mismatch between supply and demand. If the effects of adenosine are blocked with receptor antagonists, the vasodilation is also reduced. However, the effects of adenosine on the hyperemia evoked by hypercapnoea are complicated by the arousal evoked by adenosine receptor antagonists and the effects of upstream regulation. PMID- 15829182 TI - Purinergic mechanisms of the nucleus of the solitary tract and neural cardiovascular control. AB - OBJECTIVES: This review addresses the role of central purinergic receptors in the operation of the cardiovascular reflexes. METHODS: Potential physiological role of purinergic receptors operating in the nucleus of the solitary tract (NTS) was assessed via comparison of the regional patterns of hemodynamic and sympathetic responses evoked by selective stimulation/inhibition of NTS purinergic receptor subtypes, with the patterns evoked by stimulation and unloading of arterial baroreceptors, and other known patterns of autonomic responses. The effects of sino-aortic denervation plus vagotomy and ionotropic glutamatergic blockade of NTS mechanisms on the patterns of the responses were also considered. RESULTS: Selective stimulation of NTS A1 receptors with CPA evoked a pattern of regional autonomic responses consistent with inhibition of baroreflex mechanisms and facilitation/ disinhibition of chemoreflex mechanisms. Selective stimulation of NTS A(2a) receptors with CGS 21680-evoked pattern of the responses different than that evoked by stimulation of baroreflex afferents what remains in contrast to previous reports suggesting that NTS A2a receptors facilitate baroreflex transmission. The pattern of the responses was similar to that observed during hypotensive hemorrhage. Preferential, b -adrenergic iliac vasodilation evoked by stimulation of adenosine A2a receptors and preferential activation of sympathetic output to the adrenal medulla by both adenosine A1 and A2a receptors are consistent with contribution of these receptors to the defense response, stress and exercise. These observations support previous findings that NTS A1 receptors contribute to the hypothalamic defense response. The effects of stimulation and blockade of NTS P2x receptors with alpha, beta-methylene ATP and suramin, respectively, suggested that neuronally-released ATP operating via P2x receptors may be a crucial co-transmitter with glutamate in mediating baroreflex responses. DISCUSSION: The above observations strongly suggest that purinergic receptor subtypes operating in NTS circuitry are linked to specific afferent and descending mechanisms primarily integrated in the NTS. PMID- 15829183 TI - Adenosinergic mechanisms underlying recovery of diaphragm motor function following upper cervical spinal cord injury: potential therapeutic implications. AB - OBJECTIVES: In adult rats, a latent respiratory motor pathway can be pharmacologically activated with 1,3-dimethylxanthine (theophylline) to restore respiratory-related activity to a hemidiaphragm paralysed by an ipsilateral upper cervical (C2) spinal cord hemisection. The purpose of this review is to describe mechanisms that underlie theophylline-induced recovery of respiratory-related function following C2 hemisection and to underscore the therapeutic potential of theophylline therapy in spinal cord injured patients with respiratory deficits. METHODS: Theophylline mediates recovery of respiratory-related activity via antagonism of central adenosine A(1) receptors. When administered chronically, the drug restores and maintains recovered function. Since theophylline is an adenosine receptor antagonist with affinity for both the adenosine A(1) and A(2) receptors, we assessed the relative contributions of each receptor to functional recovery. While A(1) receptor antagonism plays a predominant role, activation of the A(2) receptors by specific agonists subserves the A(1) receptor-mediated actions. That is, when an adenosine A(2) receptor agonist is administered first, it primes the system such that subsequent administration of the A(1) antagonist induces a greater degree of recovered respiratory activity than when the antagonist alone is administered. RESULTS: Chronic oral administration of theophylline in C2 hemisected animals demonstrates that even when animals have been weaned from the drug, theophylline-induced recovered respiratory actions persist. This suggests that in clinical application, it may not be necessary to maintain patients on long-term theophylline. We have shown that recovery of respiratory-related activity in the ipsilateral phrenic nerve can occur spontaneously 3-4 months after C2 hemisection. Theophylline administration after this post-injury period obliterates/negates the recovery function. This indicates strongly that there is therapeutic window (more acutely after injury) for the initiation of theophylline therapy. We have also demonstrated that peripheral (carotid bodies) adenosine A(1) receptors can be selectively activated to modulate theophylline-induced CNS actions. Blocking central adenosine receptors while simultaneously activating peripheral adenosine receptors minimizes the potential of respiratory muscle fatigue with theophylline. DISCUSSION: The significance of the current findings lies in the potential clinical application of theophylline therapy in spinal cord injured patients with respiratory deficits. The ultimate goal of theophylline therapy is to wean ventilator dependent patients off ventilatory support. Thus far, our animal studies suggest that the onset of theophylline therapy must be soon after injury. PMID- 15829184 TI - Parkinson's disease risk factors: genetic, environmental, or both? AB - Perhaps one of the most important questions posed by the neurobiology of aging concerns the pathogenic mechanisms in Parkinson's disease (PD). Recently, it was suggested that exposure to pesticides could be the main cause of PD. Another study reported that the environmental endotoxin, lipopolysaccaride produced by Salmonella minnesota, might be a risk factor for PD. An alternative explanation is the genetic component, which has been suggested to be an important risk factor. Epidemiological studies have identified a positive family history of Parkinson as one of the most important risk factors for the disease. However, these studies neither examined nor reviewed the medical records of the family members. The twin study stated that the major factors in the etiology of PD are non-genetic. Meanwhile, epidemiological studies from China have shown that the prevalence of PD is much lower than in the Caucasian population, explained by the low frequency of cytochrome P-450 CYP2D6 debrisoquine hydroxylase gene polymorphism. The etiology of idiopathic PD is still a question for scientists, and calls for further research, especially with the growing proportion of elderly and the rising incidence of PD worldwide. Future research for PD risk factors should consider that multiple interactions occur in PD, resulting in a complex trait, which includes genetic, acquired, and environmental components. PMID- 15829185 TI - Ultrasonic blood flow volume assessment in the extracranial internal carotid artery in arteriovenous malformations. AB - Angle-independent ultrasound assessment of the volume flow in the extracranial internal carotid artery (ICA) is a new approach to evaluate the perfusion status of the human brain. Normal values using a new technical device (Quantix ND, Cardiosonix Ltd, Israel) and correlation analysis to quantitative CBF measurements were recently published. Aim of this study was to evaluate this device in arteriovenous malformations where cerebral blood flow is known to be increased. Five patients suffering from large supratentorial arteriovenous malformations (AVM) could be examined. All patients showed pre-operatively elevated flow volumes in the internal carotid artery on the side harbouring the pathological lesion compared with normal values evaluated in 50 volunteers. The contralateral internal carotid artery showed volume flow values within the normal limit. After surgical resection of the malformation normal values could be measured in all patients. Pathological elevated increases in cerebral blood flow can be evaluated by ultrasonic blood flow volume assessment in the extracranial internal carotid artery. Therefore, the Quantix ND seems a valuable tool in diagnosing cerebral hyperemia. PMID- 15829186 TI - Recombinant tissue plasminogen activator attenuates basal lamina antigen loss after experimental focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: The use of recombinant tissue plasminogen activator (rt PA) is a proven therapy in acute stroke. Main concerns are based on hemorrhagic complications, which are connected with microvascular integrity loss. The aim of this study was to evaluate microvascular changes after various doses of rt-PA. METHODS AND RESULTS: Focal cerebral ischemia for 3 hours was induced using the suture model in rats and followed by 24 hours of reperfusion. Six rats received either saline, 0.9, 9, or 18 mg rtPA/kg body weight at the end of ischemia. By immunostaining of collagen type IV the density of microvessels and the total stained area in the basal ganglia and cortex was measured. Comparison of the ischemic with the non-ischemic hemisphere showed significantly less reduction of the number of microvessels in rats treated with low-dose rt-PA than in the other groups: controls 17 +/- 3% (basal ganglia), 12 +/- 7% (cortex); 0.9 mg rt-PA, 18 +/- 3%, 10 +/- 4%; 9 mg, 21 +/- 4%, 13 +/- 7%; 18 mg, 22 +/- 4%, 15 +/- 8%. A similar effect was observed on the total stained area: control 25 +/- 4% (basal ganglia), 14 +/- 7% (cortex); 0.9 mg rt-PA, 23 +/- 2%, 7 +/- 4%; 9 mg, 28 +/- 4%, 15 +/- 4%; 18 mg, 29 +/- 4%, 17 +/- 5%, p<0.001. The significant reduction of the area of infarction after low and moderate doses of rt-PA was visualized with an MAP2-antibody, and the volume was calculated by 3-D reconstruction: control, 165.2 mm 3 +/- 21%; 0.9 mg rt-PA, 102.6 mm 3 +/- 16%; 9 mg, 101.2 mm 3 +/- 17%; 18 mg, 133.0 mm 3 +/- 24%; p < 0.001. CONCLUSIONS: Rats exposed to low-dose rt-PA preserved basal lamina structures, and showed smaller infarct sizes. The protective effect of low-dose rt-PA might be due to an increased microvascular patency rate. PMID- 15829187 TI - Effects of Abeta25-35 on neurogenesis in the adult mouse subventricular zone and dentate gyrus. AB - It has been demonstrated that neuorgenesis driven by neural precursor cells persists well into the adult period. This study was to observe the effects of Amyloid-beta (25-35) peptide (Abeta(25-35)) on neurogenesis in the subventricular zone and dentate gyrus of adult mouse brain. Aggregated Abeta(25-35)(1 mg/ml, 3 microl) was injected into the lateral ventricle of adult mouse. Animals were transcardially perfused with 4% paraformaldehyde in PBS, respectively at 5, 10, 20, 30 days after the Abeta(25-35) injection. All the animals were injected with BrdU (50 mg/kg, i. p) to label the neural precursor cells 24 h before the each perfusion. NeuN immunofluorescence and BrdU immunohistology were performed. It was found that Abeta(25-35) could injure the mature neurons and decrease the number of NeuN positive neurons. It also showed that Abeta(25-35) inhibited neurogenesis and significantly decreased the number of BrdU positive cells in the dentate gyrus of hippocampus, but it had no obvious effects on neurogenesis in the subventricular zone. The present results indicated that Abeta(25-35) could impair neurogenesis in the hippocampus of adult mouse brain. PMID- 15829188 TI - Emerging infectious diseases: a 10-year perspective from the National Institute of Allergy and Infectious Diseases. AB - Although optimists once imagined that serious infectious disease threats would by now be conquered, newly emerging (e.g., severe acute respiratory syndrome [SARS]), reemerging (e.g., West Nile virus), and even deliberately disseminated infectious diseases (e.g., anthrax bioterrorism) continue to appear throughout the world. Over the past decade, the global effort to identify and characterize infectious agents, decipher the underlying pathways by which they cause disease, and develop preventive measures and treatments for many of the world's most dangerous pathogens has resulted in considerable progress. Intramural and extramural investigators supported by the National Institute of Allergy and Infectious Diseases (NIAID) have contributed substantially to this effort. This overview highlights selected NIAID-sponsored research advances over the past decade, with a focus on progress in combating HIV/AIDS, malaria, tuberculosis, influenza, SARS, West Nile virus, and potential bioterror agents. Many basic research discoveries have been translated into novel diagnostics, antiviral and antimicrobial compounds, and vaccines, often with extraordinary speed. PMID- 15829189 TI - Recurring methicillin-resistant Staphylococcus aureus infections in a football team. AB - An outbreak of community-associated methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infection (SSTI) occurred in a college football team from August to September 2003. Eleven case-players were identified, and boils were the most common sign. Linemen had the highest attack rate (18%). Among 99 (93% of team) players with cultured specimens, 8 (8%) had positive MRSA nasal cultures. All available case-players' MRSA isolates characterized had the community-associated pulsed-field type USA300. A case-control study found that sharing bars of soap and having preexisting cuts or abrasions were associated with infection. A carrier-control study found that having a locker near a teammate with an SSTI, sharing towels, and living on campus were associated with nasal carriage. Successful outbreak control measures included daily hexachlorophene showers and hygiene education. PMID- 15829190 TI - Bed bug infestations in an urban environment. AB - Until recently, bed bugs have been considered uncommon in the industrialized world. This study determined the extent of reemerging bed bug infestations in homeless shelters and other locations in Toronto, Canada. Toronto Public Health documented complaints of bed bug infestations from 46 locations in 2003, most commonly apartments (63%), shelters (15%), and rooming houses (11%). Pest control operators in Toronto (N = 34) reported treating bed bug infestations at 847 locations in 2003, most commonly single-family dwellings (70%), apartments (18%), and shelters (8%). Bed bug infestations were reported at 20 (31%) of 65 homeless shelters. At 1 affected shelter, 4% of residents reported having bed bug bites. Bed bug infestations can have an adverse effect on health and quality of life in the general population, particularly among homeless persons living in shelters. PMID- 15829191 TI - Experimental infection of prairie dogs with monkeypox virus. AB - Studies of experimental infection of prairie dogs (Cynomys ludovicianus) with monkeypox virus are described. After intraperitoneal infection, all of the animals died within 11 days. Virus was cultured from their blood and oropharynx several days before death; at necropsy, most of the organs tested contained monkeypox virus. Marked hepatic and splenic necrosis were observed, along with mild inflammatory changes in the lungs. After intranasal (IN) infection, the primary pathologic changes were in the lungs and pleural cavity. Some of the IN infected animals (40%) survived, and monkeypox virus could be cultured from their nasal discharge and oropharynx for <22 days. Ulcerative lesions also developed on the lips, tongue, and buccal mucosa of the surviving animals. Our findings support an earlier report, which suggested that infected prairie dogs can transmit monkeypox virus by respiratory and mucocutaneous contact with susceptible animals and persons. PMID- 15829192 TI - Antimicrobial-resistant invasive Escherichia coli, Spain. AB - Surveillance System. A network of 32 Spanish hospitals, serving approximately 9.6 million persons, submitted antimicrobial-susceptibility data on 7,098 invasive Escherichia coli species (2001-2003). Resistance to ampicillin, cotrimoxazole, ciprofloxacin, gentamicin, and tobramycin was found at rates of 59.9%, 32.6%, 19.3%, 6.8%, and 5.3%, respectively. Resistance to multiple drugs increased from 13.8% in 2001 to 20.6% in 2003 (p <0.0001). Antimicrobial consumption data were obtained from the Spanish National Health System. In spite of decreased cephalosporin and beta-lactam use, overall extended-spectrum beta-lactamase production increased from 1.6% (2001) to 4.1% (2003) (p <0.0001), mainly due to the rising prevalence of cefotaximases. Resistance to ciprofloxacin significantly increased, mostly in community-onset infections, which coincided with a rise in community quinolone use. Cotrimoxazole resistance remained stable at approximately 30%, even though its use was dramatically reduced. PMID- 15829193 TI - Staphylococcus aureus bacteremia, Australia. AB - Staphylococcus aureus bacteremia (SAB) is common and increasing worldwide. A retrospective review was undertaken to quantify the number of cases, their place of acquisition, and the proportions caused by methicillin-resistant S. aureus (MRSA) in 17 hospitals in Australia. Of 3,192 episodes, 1,571 (49%) were community onset. MRSA caused 40% of hospital-onset episodes and 12% of community onset episodes. The median rate of SAB was 1.48/1,000 admissions (range 0.61 3.24; median rate for hospital-onset SAB was 0.7/1,000 and for community onset 0.8/1,000 admissions). Using these rates, we estimate that approximately 6,900 episodes of SAB occur annually in Australia (35/100,000 population). SAB is common, and a substantial proportion of cases may be preventable. The epidemiology is evolving, with >10% of community-onset SAB now caused by MRSA. This is an emerging infectious disease concern and is likely to impact on empiric antimicrobial drug prescribing in suspected cases of SAB. PMID- 15829194 TI - Echovirus 30, Jiangsu Province, China. AB - An outbreak of aseptic meningitis occurred in the northern area of Jiangsu Province in China from January to July in 2003. A total of 1,681 cases were involved in this outbreak, and 99% of patients were <15 years of age. To identify the etiologic agent, 66 cerebrospinal fluid specimens were tested by cell culture. Eighteen showed an enteroviruslike cytopathic effect on MRC-5 human fetal diploid lung cells. An enterovirus primer-mediated reverse transcriptase polymerase chain reaction, a standard neutralization assay, and sequencing of the complete capsid-encoding (VP1) gene identified the 18 isolates (FDJS03) as echovirus 30. At least a 10% difference was seen in nucleotide sequences of VP1 between FDJS03 isolates and other global strains of echovirus 30. Phylogenetic analysis based on complete sequences of VP1 was performed to further characterize the FDJS03 isolates. This report is the first to identify a distinct lineage of echovirus 30 as a probable cause of this outbreak. PMID- 15829195 TI - Reemergence of canine Echinococcus granulosus infection, Wales. AB - As a consequence of large-scale outdoor slaughter of sheep during the 2001 foot and mouth disease (FMD) outbreak in the United Kingdom and the possibility of increased risk for transmission of Echinococcus granulosus between sheep and dogs, a large survey of canine echinococcosis was undertaken in mid-Wales in 2002. An Echinococcus coproantigen-positive rate of 8.1% (94/1,164) was recorded on 22% of farms surveyed, which compares to a rate of 3.4% obtained in the same region in 1993. Positivity rates between FMD-affected properties and unaffected ones did not differ significantly. Significant risk factors for positive results in farm dogs were allowing dogs to roam free and the infrequent dosing (>4-month intervals) of dogs with praziquantel. When these data are compared to those of a previous pilot hydatid control program in the area (1983-1989), an increase in transmission to humans appears probable. PMID- 15829196 TI - European bat lyssavirus in Scottish bats. AB - We report the first seroprevalence study of the occurrence of specific antibodies to European bat lyssavirus type 2 (EBLV-2) in Daubenton's bats. Bats were captured from 19 sites across eastern and southern Scotland. Samples from 198 Daubenton's bats, 20 Natterer's bats, and 6 Pipistrelle's bats were tested for EBLV-2. Blood samples (N = 94) were subjected to a modified fluorescent antibody virus neutralization test to determine antibody titer. From 0.05% to 3.8% (95% confidence interval) of Daubenton's bats were seropositive. Antibodies to EBLV-2 were not detected in the 2 other species tested. Mouth swabs (N = 218) were obtained, and RNA was extracted for a reverse transcription-polymerase chain reaction (RT-PCR). The RT-PCR included pan lyssavirus-primers (N gene) and internal PCR control primers for ribosomal RNA. EBLV-2 RNA was not detected in any of the saliva samples tested, and live virus was not detected in virus isolation tests. PMID- 15829197 TI - Influenza outbreak control in confined settings. AB - Influenza may rapidly disseminate within populations living in confined settings, causing considerable morbidity and disrupting daily activities. We describe an influenza A outbreak on a military base where 3,000 young adults, most of whom were unvaccinated, lived in close daily contact. Visits to the base clinic by 48 persons with acute respiratory illness within 2 days allowed early identification of this outbreak and prompted immediate epidemiologic investigation. Overall, 85 personnel (2.83% of base population) met the case definition for influenzalike illness. On-site laboratory confirmation with field detection kits, rapid implementation of respiratory illness control protocols, and a mass vaccination campaign were applied to limit disease dissemination. The outbreak was halted 14 days after the mass vaccination campaign was completed. We review the control measures available for controlling influenza outbreaks in confined settings and discuss the role of rapid mass vaccination within this context. PMID- 15829198 TI - Buruli ulcer recurrence, Benin. AB - Buruli ulcer is a recognized public health problem in West Africa. In Benin, from 1989 to 2001, the Centre Sanitaire et Nutritionnel Gbemoten (CSNG) treated >2,500 Buruli ulcer patients. From March 2000 to February 2001, field trips were conducted in the Zou and Atlantique regions. The choice of the 2 regions was based on the distance from CSNG and on villages with the highest number of patients treated at CSNG. A total of 66 (44.0%) of 150 former patients treated at CSNG were located in the visited villages. The recurrence rate of CSNG-treated patients after a follow-up period of up to 7 years was low (6.1%, 95% confidence interval [CI] 2.0-15.6). We attribute this low rate to the high quality of Buruli ulcer treatment at an accessible regional center (CSNG). The World Health Organization definition of a Buruli ulcer recurrent case should be revised to include a follow-up period >1 year. PMID- 15829199 TI - Childhood hemolytic uremic syndrome, United Kingdom and Ireland. AB - We conducted prospective surveillance of childhood hemolytic uremic syndrome (HUS) from 1997 to 2001 to describe disease incidence and clinical, epidemiologic and microbiologic characteristics. We compared our findings, where possible, with those of a previous study conducted from 1985 to 1988. The average annual incidence of HUS for the United Kingdom and Ireland (0.71/100,000) was unchanged from 1985 to 1988. The overall early mortality had halved, but the reduction in mortality was almost entirely accounted for by improved outcome in patients with diarrhea-associated HUS. The principal infective cause of diarrhea-associated HUS was Shiga toxin-producing Escherichia coli O157 (STEC O157), although in the 1997 2001 survey STEC O157 phage type (PT) 21/28 had replaced STEC O157 PT2 as the predominant PT. The risk of developing diarrhea-associated HUS was significantly higher in children infected with STEC O157 PT 2 and PT 21/28 compared with other PTs. Hypertension as a complication of HUS was greatly reduced in patients with diarrhea-associated HUS. PMID- 15829200 TI - Tuberculosis elimination in the Netherlands. AB - This study assessed progress towards tuberculosis (TB) elimination in the Netherlands by using DNA fingerprinting. Mycobacterium tuberculosis strains were defined as new if the IS6110 restriction fragment length polymorphism pattern had not been observed in any other patient during the previous 2 years. Other cases were defined as clustered and attributed to recent transmission. In the period 1995-2002, the incidence of TB with new strains was stable among non-Dutch residents and declined among the Dutch. However, the decline among the Dutch was restricted to those >or=65 years of age. Moreover, the average number of secondary cases per new strain did not change significantly over time. We conclude that the decline of TB in the Netherlands over the past decade was mainly the result of a cohort effect: older birth cohorts with high infection prevalence were replaced by those with lower infection prevalence. Under current epidemiologic conditions and control efforts, TB may not be eliminated. PMID- 15829201 TI - Epidemiology of Escherichia coli O157:H7 outbreaks, United States, 1982-2002. AB - Escherichia coli O157:H7 causes 73,000 illnesses in the United States annually. We reviewed E. coli O157 outbreaks reported to Centers for Disease Control and Prevention (CDC) to better understand the epidemiology of E. coli O157. E. coli O157 outbreaks (>or=2 cases of E. coli O157 infection with a common epidemiologic exposure) reported to CDC from 1982 to 2002 were reviewed. In that period, 49 states reported 350 outbreaks, representing 8,598 cases, 1,493 (17%) hospitalizations, 354 (4%) hemolytic uremic syndrome cases, and 40 (0.5%) deaths. Transmission route for 183 (52%) was foodborne, 74 (21%) unknown, 50 (14%) person to-person, 31 (9%) waterborne, 11 (3%) animal contact, and 1 (0.3%) laboratory related. The food vehicle for 75 (41%) foodborne outbreaks was ground beef, and for 38 (21%) outbreaks, produce. PMID- 15829202 TI - Web-based investigation of multistate salmonellosis outbreak. AB - We investigated a large outbreak of Salmonella enterica serotype Javiana among attendees of the 2002 U.S. Transplant Games, including 1,500 organ transplant recipients. Web-based survey methods identified pre-diced tomatoes as the source of this outbreak, which highlights the utility of such investigative tools to cope with the changing epidemiology of foodborne diseases. PMID- 15829203 TI - Ross River virus disease reemergence, Fiji, 2003-2004. AB - We report 2 clinically characteristic and serologically positive cases of Ross River virus infection in Canadian tourists who visited Fiji in late 2003 and early 2004. This report suggests that Ross River virus is once again circulating in Fiji, where it apparently disappeared after causing an epidemic in 1979 to 1980. PMID- 15829204 TI - Mycobacteria in nail salon whirlpool footbaths, California. AB - In 2000, an outbreak of Mycobacterium fortuitum furunculosis affected customers using whirlpool footbaths at a nail salon. We swabbed 30 footbaths in 18 nail salons from 5 California counties and found mycobacteria in 29 (97%); M. fortuitum was the most common. Mycobacteria may pose an infectious risk for pedicure customers. PMID- 15829205 TI - Coxiella burnetii in bulk tank milk samples, United States. AB - Dairy cattle are a primary reservoir of Coxiella burnetii, which causes Q fever. However, no recent nationwide studies have assessed the prevalence and risks of Q fever in dairy cattle. We report >or=94% prevalence in samples of bulk tank milk from U.S. dairy herds tested during the past 3 years. PMID- 15829206 TI - Human spotted fever rickettsial infections. AB - Serum specimens from patients at 4 sites in Peru were tested for evidence of spotted fever group rickettsial infection. Results showed that 30 (18%) of 170 patients had spotted fever group rickettsial infections, which likely caused their illnesses. These findings document laboratory-confirmed spotted fever from diverse areas of Peru. PMID- 15829207 TI - Patient contact recall after SARS exposure. AB - We reinterviewed healthcare workers who had been exposed to a patient with severe acute respiratory syndrome (SARS) in an intensive care unit to evaluate the effect of time on recall reliability and willingness to report contact activities and infection control precautions. Healthcare workers reliably recalled events 6 months after exposure. PMID- 15829208 TI - Molecular mechanisms of West Nile virus pathogenesis in brain cell. AB - We analyzed the response of human glioma cells to West Nile virus infection by investigating host transcriptional changes. Changes in expression of 23 genes showed similarities to those in other neurodegenerative diseases. These changes may be useful as potential biomarkers and elucidate novel mechanisms behind the neuropathology of infection with this virus. PMID- 15829209 TI - The future of foot and ankle care: training the next generation. PMID- 15829210 TI - Long-term results of arthroscopic ankle arthrodesis. AB - BACKGROUND: More than 40 open procedures have been described for ankle arthrodesis, most with high complication rates. Since its description in 1983, arthroscopic ankle arthrodesis has become a viable option in selected patients. With one of the largest series in the literature, the purpose of the paper was to analyze the results of arthroscopic ankle arthrodesis at our institution. METHODS: Between 1989 and 2002, 35 patients with end-stage ankle arthritis underwent arthroscopic ankle arthrodesis. The average followup was 72 months, with a range of 24 to 167 months. Patients returned for a clinical and radiolographic evaluation using the grading systems of Mazur and Morgan. All patients had preoperative and postoperative radiographic evaluation to assess fusion. Indications for arthroscopic ankle arthrodesis included failure of at least 6 months of conservative treatment, minimal or mild correctable deformity in the coronal plane, and no active infections. RESULTS: The overall fusion rate was 97% (34 of 35 patients). The average time to fusion was 11.8 weeks, with a range of 8 to 18 weeks. There were 74% good to excellent results by the Mazur grading system and 83% by the Morgan system. There were no infections or neurovascular injuries. In the three patients who required bone stimulators for delayed unions, fusion occurred in two, and one had a nonunion. Eleven patients had screws removed because of pain at an average of 11 months after the initial surgery. CONCLUSION: Our study demonstrated a high fusion rate with minimal complications for arthroscopic ankle arthrodesis. PMID- 15829211 TI - Peroneal tendon lesions caused by antiglide plates used for fixation of lateral malleolar fractures: the effect of plate and screw position. AB - BACKGROUND: Posterolateral antiglide plating of unstable AO-type B lateral malleolar fractures is biomechanically stronger than lateral plating and causes less wound healing problems and less frequent hardware removal. However, the distal end of the plate or the screws may cause peroneal tendinitis. The limits of safe hardware placement have not been established. METHOD: A retrospective analysis of 70 patients was done to determine hardware position and identify peroneal tendon lesions. An adjunct study involved dissection of the retromalleolar region in 10 embalmed cadaver specimens to study the anatomy of the osteosynovial peroneal groove. RESULTS: Thirty of 70 (43%) patients had the plate removed because of discomfort or signs of peroneal tendinitis. Peroneal tendon lesions were identified intraoperatively in nine of the 30 (30%) patients. Only two of these nine patients had felt symptoms preoperatively. Placement of the distal end of the plate distal to the proximal third of the lateral malleolus did not correlate with a peroneal tendon lesion. However, this placement combined with a screw in the most distal hole of the plate and a prominent screw head was strongly correlated with peroneal tendon lesions. In the anatomic specimens the shape of the osteosynovial part of the peroneal groove was uniform, but its length showed greater variation than the length of the foot. CONCLUSIONS: Antiglide plating of lateral malleolar fractures led to high rates of hardware removal and peroneal tendon lesions. Correlations were found to low placement of the plate together with a protruding screw head in the most distal hole of the plate. Distal screw placement should therefore be avoided or the hardware should be removed early. Absence of subjective signs of peroneal tendon irritation does not exclude even a major tendon lesion. PMID- 15829212 TI - Isokinetic strength and endurance after percutaneous and open surgical repair of Achilles tendon ruptures. AB - BACKGROUND: Reports on complete spontaneous Achilles tendon ruptures and associated treatment have become more frequent in the literature in the past two decades, as has the request for treatments that enable the finest possible functional recovery. The best available treatment is a matter of considerable controversy in the literature. The purpose of this study was to compare the isokinetic strength and endurance of the plantarflexor muscle-tendon unit in subjects who sustained rupture of the Achilles tendon and underwent either open surgery or closed percutaneous repair of the Achilles tendon. METHODS: Twenty patients (18 males, 2 females) with spontaneous ruptures of the Achilles tendon were included in this study. Ten patients were treated by open surgery, and 10 patients were treated percutaneously. All patients had ruptured their Achilles tendon more than 6 months before the study, and all of the ruptures occurred 3.5 years or less before the day of the testing. All patients underwent an oriented physical examination. An isokinetic Biodex dynamometer (Biodex Medical System, Shirley, NY) was used to measure ankle joint angle, and in plantarflexion to calculate the torque at the ankle joint (Newton/meter), and the average work (jouls) for both maximal power and endurance. Each measurement was compared to the normal ankle. RESULTS: Biodex dynamometer evaluations at 90 deg/sec demonstrated a significant difference of maximal voluntary plantarflexor torque, endurance performance and range of motion at the ankle joint between the involved and uninvolved sides in patients treated by either mode of treatment. Yet, no statistically significant differences were revealed for the parameters mentioned above between the subjects that were treated either percutaneously or by an open surgery. CONCLUSIONS: In functional terms, the biomechanical outcomes of open surgery and percutaneous repair for acute ruptures of the Achilles tendon are both effective. PMID- 15829213 TI - Tenosynovitis of the flexor hallucis longus: a clinical study of the spectrum of presentation and treatment. AB - BACKGROUND: Symptoms associated with flexor hallucis longus (FHL) pathology can manifest themselves anywhere along its length from the posterior leg to the plantar foot and the hallux. This study describes the spectrum of clinical presentations seen with FHL pathology, illustrates the relevant physical examination findings, and outlines a treatment approach. MATERIALS: Computerized medical data was prospectively collected on 81 patients treated between January, 1997 and March, 2002. The 55 females and 26 males had an average age of 38.3 years, with a mean follow-up of 21.3 months. Forty-five of 81 had previous therapy that failed, usually for "plantar fasciitis." Twenty-seven were active athletically and 24 related the onset of symptoms to a specific traumatic episode. Pain was located at the posteromedial ankle in 40, plantar heel in 23, plantar midfoot in 22, and multiple locations in 16. All patients had tenderness of the FHL. Restriction of FHL excursion was demonstrated in 30 patients by limited hallux metatarsophalangeal joint dorsiflexion when the ankle was dorsiflexed ("FHL stretch test"). Thirty-four patients had magnetic resonance imaging of the FHL, 28 (82%) of which were positive for synovitis of the FHL. Treatment included an FHL stretching program, short-term immobilization, and operative decompression and synovectomy in patients for whom nonoperative treatment failed. RESULTS: Of the 58 patients treated nonoperatively, 37 (64%) had successful results. Twenty-three patients had surgery, 20 at the posterior ankle fibro-osseous tunnel, and three in the sesamoid region. All patients treated operatively had successful outcomes. A subset of 10 patients had hallux rigidus symptoms without significant osteophyte formation. All 10 obtained successful results with treatment directed at restoring normal FHL excursion (nine nonoperatively, one by FHL release). This suggests that limited FHL excursion may be an etiology for the development of hallux rigidus. CONCLUSIONS: Clinical syndromes related to the FHL are more frequent than previously reported. The close relationship of the FHL to commonly injured structures (such as the plantar fascia) contributes to significant delays in effective treatment. PMID- 15829214 TI - Tibiotalocalcaneal arthrodesis with a retrograde intramedullary nail: a biomechanical analysis of the effect of nail length. AB - BACKGROUND: Fatigue fractures of the tibia have been observed at the level of the proximal end of the nail after successful tibiocalcaneal arthrodesis with a retrograde intramedullary device. MATERIALS: To study the effect of nail length, five matched pairs of cadaver tibiae were instrumented with strain gauges and potted in methylmethacrylate from a level 3 cm proximal to the distal medial malleolus to simulate a successful tibiocalcaneal arthrodesis. A standard length (15 cm) ankle arthrodesis nail and an identical longer device terminating in the proximal tibial metaphysis were inserted in each paired tibia using appropriate technique. The strain of the posterior cortex of the tibia was recorded under bending moments of up to 50 Nm for each intact specimen after nail insertion and after proximal locking of the nail. The nails were then exchanged between the specimens of the same pairs and the experiment was repeated to insure uniformity. RESULTS: The standard length locked nail increased the principal strain of the posterior cortex of the tibia at the level of the proximal screw holes 5.3 times more than the locked long nail (353 and 67 microstrains), respectively. This stress concentration was not observed when the proximal extent of the nail terminated within the proximal tibial metaphysis. CONCLUSION: A successful tibiocalcaneal arthrodesis with a standard length locked intramedullary nail creates stress concentration around the proximal screw holes that may be responsible for the fractures observed clinically. This study supports the use of a "long" retrograde locked intramedullary nail for tibiocalcaneal arthrodesis in patients with systemic or localized osteopenia. PMID- 15829215 TI - A comparison of plates with and without locking screws in a calcaneal fracture model. AB - BACKGROUND: We compared different plates in an experimental calcaneal fracture model under biocompatible loading. METHODS: Four plates were tested: a plate without locked screws (Synthes), and three different plates with locked screws (Newdeal, Darco, Synthes). Synthetic calcanei (Sawbone) were osteotomized to create a fracture model, and the plates were fixed onto them. Seven specimens for each plate model were subjected to cyclic loading (preload 20 N, 1,000 cycles with 800 N, 0.75 mm/s), and load to failure (0.75 mm/s). Motion, forces, plastic deformation of the plate, and consequent depression of the posterior joint facet were analyzed. RESULTS: During cyclic loading, all plates with locked screws showed statistically significant lower displacement in the primary loading direction than the plates without locked screws. Mean values (mm) of maximal displacements for each plate during cyclic loading were as follows: Synthes, 3.5; Darco, 4.5; Newdeal, 5.0; Synthes without locked screws, 7.5; (p < 0.001). No statistically significant differences between the plates were found in relation to loads to failure and corresponding displacement. CONCLUSION: This is the first biomechanical study to assess the stability of different plates currently in use in our practice for the fixation of calcaneal fractures. Our results showed that plates with locked screws provided greater stability during cyclic loading than the plate without locked screws. PMID- 15829216 TI - Review of proximal interphalangeal joint excisional arthroplasty for the correction of second hammer toe deformity in 100 cases. AB - BACKGROUND: The main bony procedures used in the treatment of second hammertoe are excisional arthroplasty and arthrodesis of the proximal interphalangeal (PIP) joint. While a number of studies have reported the outcomes after PIP joint arthrodesis, there are few reports of the outcome of excisional arthroplasty of the PIP joint for the correction of second hammertoes. The purpose of this study was to evaluate the long-term outcome of PIP joint excisional arthroplasty. METHODS: Seventy-five patients (100 toes) who had excisional arthroplasty of the PIP joint for the correction of second hammertoe were reviewed at an average followup of 44 months. All patients were seen in clinic and were physically examined to assess the alignment and function of each digit. The radiographs were evaluated preoperatively. The AOFAS clinical rating scale was used preoperatively and at final followup. Patients were asked to rate their satisfaction on a scale of 0 to 10 and were asked if they would be happy to have the surgery under similar circumstances again. RESULTS: The mean preoperative AOFAS clinical rating scale was 46. At final followup this increased to 94, showing an average improvement of 48 points (p < 0.0001). The mean satisfaction on a scale of 0 to 10 (with 0 equaling complete dissatisfaction and 10 complete satisfaction) was 9.3 (SD 1.3). One patient would not have been happy to undergo outpatient surgery again, because of a change in her social circumstances. CONCLUSION: In a group of 75 patients who had PIP joint arthroplasty for the correction of second hammertoe, high levels of satisfaction were achieved. No serious complications were encountered, and revision surgery was required in just two cases. Floating toe was the most common complication encountered, and although this did decrease the level of patient satisfaction in seven patients it was not a cause of footwear irritation or pain in any patient. PMID- 15829217 TI - Dislocation and relocation of the tarsometatarsal joint as an approach for resection of the deep plantar fibromatosis: case report. PMID- 15829218 TI - Tuberculous tenosynovitis of the anterior tibial and extensor hallucis longus tendons: case report. PMID- 15829219 TI - Current topic review: subtalar arthroereisis for the correction of flexible flatfoot. PMID- 15829220 TI - Melanin concentrating hormone (MCH): a novel neural pathway for regulation of GnRH neurons. AB - The link between the state of energy balance and reproductive function is well known. Thus, signals denoting negative energy balance and the accompanying hyperphagic drive are likely to be factors in the suppression of gonadotropin releasing hormone (GnRH) activity. We have previously found that appetite regulating systems, such as neuropeptide Y (NPY) in the arcuate nucleus (ARH) and orexin in the lateral hypothalamic area (LHA), send fiber projections that come in close apposition with GnRH neurons. Furthermore, the appropriate receptors, NPY Y5 and OR-1, respectively, are coexpressed on GnRH neurons, providing neuroanatomical evidence for a direct link between the NPY and orexin systems and GnRH neurons. Therefore, these orexigenic neuropeptide systems are potential candidates that convey information about energy balance to GnRH neurons. The current studies focused on melanin concentrating hormone (MCH), another orexigenic neuropeptide system located in the LHA that is sensitive to energy balance. The results showed that MCH fiber projections came in close apposition with approximately 85-90% of GnRH cell bodies throughout the preoptic area and anterior hypothalamic area in the rat. In addition, the MCH receptor (MCHR1) was coexpressed on about 50-55% of GnRH neurons. These findings present evidence for a possible direct neuroanatomical pathway by which MCH may play a role in the regulation of GnRH neuronal function. Thus, MCH is another potential signal that may serve to integrate energy balance and reproductive function. PMID- 15829221 TI - Granulocyte colony-stimulating factor induces sensorimotor recovery in intracerebral hemorrhage. AB - Granulocyte colony-stimulating factor (G-CSF) has been used in the treatment of neutropenia in hematologic disorders. The neuroprotective and anti-inflammatory effects of G-CSF were reported in various neurological disease models. In this study, we examined whether G-CSF induces functional recovery after intracerebral hemorrhage (ICH). ICH was induced using collagenase injection in adult rats. Either G-CSF (50 microg/kg, i.p.) or saline was given from 2 h after ICH and every 24 h for 3 days. 72 h after ICH induction, the rats were sacrificed for histological analysis and measurement of brain edema. Behavioral tests were performed before and 1, 7, 14, 21, 28, and 35 days after ICH. We also measured the blood-brain barrier (BBB) permeability using Evans blue dye injection method. G-CSF-treated rats recovered better on rotarod and limb placing tests, starting from 14 days throughout 5 weeks after ICH. The brain water content and BBB permeability of G-CSF-treated group decreased in the lesioned hemispheres compared with those of ICH-only group. In G-CSF-treated group, the number of TUNEL+, myeloperoxidase+, and OX42+ cells was smaller than that of ICH-only group in the periphery of hematoma. These findings suggest that G-CSF induces long-term sensorimotor recovery after ICH with reduction of brain edema, inflammation, and perihematomal cell death. PMID- 15829223 TI - Galanin and perseveration. AB - Galanin is a 29/30 amino acid neuropeptide that has been shown to impair learning and memory task performance and also have roles in somatosensation, stress responses, sexual behavior, and feeding regulation. However, little is known about galanin involvement in higher cognitive processes, especially executive processes. Perseveration is a classic sign of frontal cortex damage and failure of executive control. Galanin has been shown to disrupt the performance of maze delayed alternation tasks and the operant, spatial delayed nonmatch-to-position (DNMTP) working memory task, tests especially sensitive to perseverative responding. To better understand this potential involvement of galanin in executive control, the present study tested the hypothesis that galanin induces perseveration. The first experiment examined the effects of galanin (10, 20 microg i.c.v.) on the performance of a simple operant response alternation task in which stimuli were assigned to one of two spatially distinct locations to produce extended sequences of presentations to one location, separated by a 10-s intertrial interval. The second experiment looked at the effects of galanin (5, 20 microg i.c.v.) on the performance of non-delayed match-to-position and nonmatch-to-position conditional discrimination operant tasks in which a minimal 1.0 s time interval separated responses. Finally, the effects of galanin (10, 20 microg i.c.v.) on delayed match-to-position (DMTP) performance were examined to determine whether response alternation (i.e., nonmatching) was critical to observing a galanin-induced impairment in this task. Galanin reduced the rate of trial completion in all the tasks, but did not alter simple or conditional discrimination accuracy. Galanin (10 microg) impaired DMTP performance in a delay independent manner. Together, these data suggest that galanin does not produce perseveration, but are consistent with a galanin-induced decrease in reinforcer strength. PMID- 15829222 TI - Chronic nicotine and dizocilpine effects on regionally specific nicotinic and NMDA glutamate receptor binding. AB - Chronic nicotine administration has long been known to increase the number of high-affinity alpha4beta2 nicotinic receptors with lesser effects on low-affinity alpha7 nicotinic receptors. Nicotine has been shown to promote the release of a variety of neurotransmitters including glutamate. Nicotine may also interact directly with the glutamatergic receptors. Nicotinic-glutamate interactions may be critical to the long-term effects of nicotine. Conversely, glutamatergic drugs may interact with the nicotinic system. Such interactions have important implications in interpretation of the mechanism of drug actions, especially when the drugs are given together. The current study examined the effects of chronic administration of nicotine (5 mg of the nicotine base/kg/day for 28 days), dizocilpine (MK-801) (0.3 mg/kg/day for 28 days), an NMDA receptor antagonist, as well as the combination of the two drugs on nicotinic and NMDA receptor densities in discrete brain regions. The chronic dose of dizocilpine used was behaviorally active causing a dramatic reduction in prepulse inhibition (PPI) of acoustic startle response. The nicotine dose used did not significantly affect PPI but previously we have found it to be behaviorally active in improving working memory function. High-affinity nicotinic receptor binding, as has been seen previously, was significantly increased by chronic nicotine in most areas. Chronic dizocilpine alone did not affect high-affinity nicotinic receptor binding, but it did modify the effects of chronic nicotine, attenuating nicotine-induced increases in the frontal cortex and striatum. Low-affinity nicotinic binding was significantly increased by chronic nicotine in only one area, the cerebellum. Chronic dizocilpine significantly increased low-affinity nicotinic binding in several brain areas, the colliculi, hippocampus, and the hypothalamus. The combination of nicotine and dizocilpine attenuated the effects of each with diminished nicotine-induced increased nicotinic low-affinity binding in the cerebellum and diminished dizocilpine-induced increased nicotinic low-affinity binding in the hippocampus and hypothalamus. In contrast, chronic nicotine and dizocilpine had a mutually potentiating effect of increasing nicotinic low affinity binding in the frontal cortex. NMDA receptor binding was affected only in the hippocampus, where both dizocilpine and nicotine significantly increased binding. Chronic nicotine effects on receptor regulation are significantly affected by concurrent blockade of NMDA glutamate receptors. PMID- 15829224 TI - Beneficial effects of PJ34 and INO-1001, two novel water-soluble poly(ADP-ribose) polymerase inhibitors, on the consequences of traumatic brain injury in rat. AB - Traumatic brain injury produces peroxynitrite, a powerful oxidant which triggers DNA strand breaks, leading to the activation of poly(ADP-ribose)polymerase-1 (PARP-1). We previously demonstrated that 3-aminobenzamide, a PARP inhibitor, is neuroprotective in a model of traumatic brain injury induced by fluid percussion in rat, suggesting that PARP-1 could be a therapeutic target. In order to confirm this hypothesis, we investigated the effects of PJ34 and INO-1001, two PARP inhibitors from structural classes other than benzamide, on the post-traumatic consequences. Pre- and post-treatments with PJ34 (30 mg/kg/day) and INO-1001 (10 mg/kg/day) decrease the neurological deficit at 3 days post-injury and this deficit is still reduced at 7 days. These neurological recovery-promoting effects are associated with the inhibition of PARP-1 activation caused by trauma, as demonstrated by abolishment of immunostaining of poly(ADP-ribose). Thus, the present work strengthens strongly the concept that PARP-1 inhibition may be a suitable approach for the treatment of brain trauma. PMID- 15829225 TI - Neurturin, persephin, and artemin in the human pre- and full-term newborn and adult hippocampus and fascia dentata. AB - The immunochemical occurrence and localization of the Glial cell line-derived neurotrophic factor (GDNF) family ligands neurturin (NTN), persephin (PSP), and artemin (ART) is described in the human postmortem hippocampus and fascia dentata from subjects aged 21 weeks of gestation to 88 years. The detectability of NTN, PSP, and ART is shown in the rat by Western blot and immunohistochemistry up to 70 h postmortem. In the human tissue, labeled neuronal perikarya were detectable for each trophin at all examined ages, with prevalent localization in the pyramidal layer of the Ammon's horn and hilus and granular layer of the fascia dentata. In the adult subjects, punctate elements were also present. Comparison of the pattern of immunoreactive structures among young and adult subjects suggests that intracellular distribution and/or trafficking of the GDNF family ligands may undergo age-related changes. Labeled glial elements were also identifiable. Western blot analysis indicates that the availability of the dimeric and monomeric forms of the trophins may vary with age and postmortem delay. The results obtained suggest the involvement of NTN, PSP, and ART in processes subserving both the organization of this cortical region during development and the functional activity and maintenance of the mature human hippocampal neurons. PMID- 15829226 TI - Detection of beta-endorphin in the cerebrospinal fluid after intrastriatal microinjection into the rat brain. AB - We have investigated to what extent microinjected beta-endorphin could migrate from the rat brain parenchyma into the CSF compartment. Exogenous rat beta endorphin (0.1 nmol) was microinjected into the left striatum 1 mm from the lateral ventricle in anesthetized male rats. CSF samples were collected at different time points up to 2 h post-injection from a catheter affixed to the atlanto-occipital membrane of the cisterna magna. Radioimmunoassay and mass spectrometry were performed on the CSF samples, and brain sections were immunostained for beta-endorphin and mu-opioid receptors. The beta-endorphin injected rats showed a marked increase in beta-endorphin immunoreactive (IR) material in the CSF, with a peak at 30-45 min post-injection, and this beta endorphin-IR material existed mainly as the intact beta-endorphin peptide. The immunohistochemistry results revealed the appearance of distinct beta-endorphin IR cell bodies in the globus pallidus and the bed nucleus of stria terminalis supracapsular part, regions distant from the injection site, at 2 h post injection of exogenous beta-endorphin. The beta-endorphin-IR in several of the globus pallidus cell bodies colocalized with the mu-opioid receptor-IR at the cell surface. These findings show that upon delivery of synthetic beta-endorphin, there is a significant intracerebral spread of the injected peptide, reaching regions far from the site of injection via diffusion in the extracellular space and flow in the cerebrospinal fluid. This may be of relevance when interpreting studies based on intracerebral injections of peptides, and advances our knowledge regarding the migration of compounds within the brain. PMID- 15829227 TI - Cerebellar thalamic activity in the macaque monkey encodes the duration but not the force or velocity of wrist movement. AB - The way in which the cerebellum influences the output of the motor cortex is not known. The aim of this study was to establish whether information about force, velocity or duration of movement is encoded in cerebellar thalamic discharge and could therefore be involved in the modulation of motor cortical activity. Extracellular single cell recordings were made from the cerebellar thalamus (66 neurones) and VPLc (49 neurones) of four conscious macaques performing simple wrist movements with various load and gain conditions imposed. A significant correlation (Spearman's; P<0.05) was found between movement duration and the duration of neuronal discharge of most cerebellar thalamic neurones (65%), the velocity of movement and rate of neuronal discharge of some cerebellar thalamic neurones (23%), but not between force of movement and rate of neuronal discharge of any cerebellar thalamic neurones. Similar relationships were found between the activity of VPLc neurones and these movement parameters. The strength of the correlations increased when many cells were grouped and analysed as an ensemble, suggesting that populations of cerebellar thalamic (and VPLc) neurones can encode a signal with higher fidelity than single neurones alone. The ensemble data confirmed that the most robust association was between the duration of neuronal discharge and movement duration. We propose that the cerebellum does not provide the motor cortex with specific information about movement force or velocity, but rather that its major role is in activating many motor cortical regions for a specific duration, thus influencing the timing of complex movements involving many muscles and joints. PMID- 15829228 TI - Effect of hypoxanthine on Na+,K+-ATPase activity and some parameters of oxidative stress in rat striatum. AB - The main objective of this study was to investigate the effects of preincubation of rat striatum homogenate in the presence of hypoxanthine, a metabolite accumulated in Lesch-Nyhan disease, on Na+,K+-ATPase activity and on some parameters of oxidative stress namely thiobarbituric acid-reactive substances (TBA-RS), total radical-trapping antioxidant parameter (TRAP) and membrane protein thiol content. Results showed that hypoxanthine significantly increased TBA-RS and reduced Na+,K+-ATPase activity, TRAP and membrane protein thiol content. In addition, we also evaluated the effect of glutathione, trolox, allopurinol and Nvarpi-nitro-L-arginine methyl ester (L-NAME) on the inhibitory effect of hypoxanthine on Na+,K+-ATPase activity in the same rat cerebral structure. All tested compounds per se did not alter Na+,K+-ATPase activity, but only glutathione and trolox prevented the effect of hypoxanthine on the enzyme activity. The effect of glutathione and trolox on hypoxanthine-induced increase of TBA-RS levels was also investigated. These antioxidants alone or combined with hypoxanthine reduced TBA-RS levels. Our present findings show that hypoxanthine induces oxidative stress in rat striatum and that the inhibition of Na+,K+-ATPase activity caused by this oxypurine was probably mediated by reactive oxygen species. It is presumed that these results might be associated with the neuronal dysfunction of patients affected by Lesch-Nyhan disease. PMID- 15829229 TI - Expression of mRNA for four subtypes of the proteinase-activated receptor in rat dorsal root ganglia. AB - Proteinase-activated receptors (PARs) are members of the superfamily of G-protein coupled receptors that initiate intracellular signaling by the proteolytic activity of extracellular serine proteases. Three member of this family (PAR-1, PAR-3, and PAR-4) are considered thrombin receptors, whereas PAR-2 is activated by trypsin and tryptase. Recently, activation of PAR-2 signal was identified as a pro-inflammatory factor that mediates peripheral sensitization of nociceptors. Activation of PAR-1 in the periphery is also considered to be a neurogenic mediator of inflammation that is involved in peptide release. Here, we investigated the expression of these four members of PARs in the adult rat dorsal root ganglia (DRG) using radioisotope-labeled in situ hybridization histochemistry. We detected mRNA for all subtypes of PARs in the DRG. Histological analysis revealed the specific expression patterns of the PARs. PAR 1, PAR-2, and PAR-3 mRNA was expressed in 29.0+/-4.0%, 16.0+/-3.2%, and 40.9+/ 1.3% of DRG neurons, respectively. In contrast, PAR-4 mRNA was mainly observed in non-neuronal cells. A double-labeling study of PARs with NF-200 and alpha calcitonin gene-related peptide (CGRP) also revealed the distinctive expression of PARs mRNA in myelinated or nociceptive neurons. This study shows the precise expression pattern of PARs mRNA in the DRG and indicates that the cells in DRG can receive modulation with different types of proteinase-activated receptors. PMID- 15829230 TI - Phosphodiesterase 4B (PDE4B) and cAMP-level regulation within different tissue fractions of rat hippocampal slices during long-term potentiation in vitro. AB - Molecular events associated with mnemonic processes and neuronal plasticity are postulated to result in functional changes in synaptic structure. One possible site is the post-synaptic density, where activity-dependent changes modulate signal transduction cascades. In this report, we detail spatial-temporal changes for phosphodiesterase 4B (PDE4B) proteins and their substrate cAMP within three neuronal fractions during early and late long-term potentiation (LTP). The cAMP dependent protein kinase A cascade--which can be regulated by distinct PDE4B activity--is required for mnemonic processes as well as mechanisms of neuronal plasticity, such as those during the maintenance or late-LTP. Fluorescence in situ hybridization studies (FISH) identified no translocation of PDE4B3 from the soma after late-LTP induction indicating a subtle, local control of PDE4B activity. Protein changes were detected within the PSD-enriched fraction. From these results, we conclude that either the changes in PDE4B are due to modulation of pre-existing mRNA, or that the protein is specifically translocated to activated synaptic structures. Furthermore, we report late changes in cAMP levels in the somato-dendritic fraction and discuss this result with the increased PDE4B1/3 doublet in the PSD-enriched fraction. PMID- 15829231 TI - Changes in hippocampal IL-15, related cytokines, and neurogenesis in IL-2 deficient mice. AB - Previous studies have demonstrated that interleukin-2 knockout (KO) mice exhibit alterations in hippocampal cytoarchitecture. Several lines of evidence suggest that these variations may result from immune dysregulation and/or autoimmunity. Thus, this study sought to compare adult IL-2 KO mice and wild-type littermates (8-12 weeks of age), the age where differences in hippocampal cytoarchitecture have previously been observed, for differences in measures of neuroimmunological status in the hippocampus. Furthermore, because IL-15 shares the same receptor subunits for signal transduction as IL-2 (IL-2/15Rbeta and gammac) that are enriched in the hippocampus and may induce inflammatory processes in IL-2 KO mice, we sought to test the hypothesis that IL-15 is elevated in the hippocampus of IL-2 KO mice. Compared to wild-type mice, IL-2 KO mice exhibited increased hippocampal protein concentrations of IL-15 as well as IL-12, IP-10, and MCP-1. These cytokine changes, however, did not correlate with levels in the peripheral circulation, and there were no T cells or an increase in MHCII-positive microglia in the hippocampus of IL-2 KO mice. Since elevated levels of certain inflammatory cytokines may impair hippocampal neurogenesis, we also tested the hypothesis that changes in neuroimmunological status would be associated with reductions in neurogenesis of neurons in the dentate gyrus of IL-2 KO mice. Contrary to this hypothesis, compared to wild-type mice, male IL-2 KO mice exhibited increased neurogenesis in both the infrapyramidal and suprapyramidal limbs of the granule cell layer of the dentate gyrus, differences that were not observed between females. These findings indicate that IL-2 gene deletion alters the neuroimmunological status of the mouse hippocampus through a dysregulation of cytokines produced by CNS cells, and in males, these changes are associated with increased hippocampal neurogenesis. PMID- 15829233 TI - Single nucleotide polymorphisms (SNPs): detection, interpretation, and application. PMID- 15829232 TI - Distribution of orexin/hypocretin immunoreactivity in the nervous system of the green Treefrog, Hyla cinerea. AB - We examined the distribution of orexin/hypocretin immunoreactive neurons and projections throughout the brain of the green treefrog (Hyla cinerea). Orexin A and B neurons were located in a single population centered on the suprachiasmatic nucleus. Orexin A and B fibers were visible across the brain, with the highest density within the preoptic area and hypothalamus. Our data suggest different distributions of orexin neurons but not projections between families of amphibians. PMID- 15829234 TI - Mass-spectrometry DNA sequencing. AB - Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been explored widely for DNA sequencing. Compared to gel electrophoresis based sequencing systems, mass spectrometry produces very high resolution of sequencing fragments, rapid separation on microsecond time scales, and completely eliminates compressions associated with gel-based systems. While most of the research efforts have focused on using mass spectrometers to analyze the DNA products from Sanger sequencing or enzymatic digestion reactions, the read lengths attainable are currently insufficient for large-scale de novo sequencing. The advantage of mass-spectrometry sequencing is that one can unambiguously identify frameshift mutations and heterozygous mutations making it an ideal choice for resequencing projects. In these applications, DNA sequencing fragments that are the same length but with different base compositions are generated, which are challenging to consistently distinguish in gel-based sequencing systems. In contrast, MALDI-TOF MS produces mass spectra of these DNA sequencing fragments with nearly digital resolution, allowing accurate determination of the mixed bases. For these reasons mass spectrometry based sequencing has mainly been focused on the detection of frameshift mutations and single nucleotide polymorphisms (SNPs). More recently, assays have been developed to indirectly sequence DNA by first converting it into RNA. These assays take advantage of the increased resolution and detection ability of MALDI-TOF MS for RNA. PMID- 15829235 TI - Advances in sequencing technology. AB - Faster sequencing methods will undoubtedly lead to faster single nucleotide polymorphism (SNP) discovery. The Sanger method has served as the cornerstone for genome sequence production since 1977, close to almost 30 years of tremendous utility [Sanger, F., Nicklen, S., Coulson, A.R, DNA sequencing with chain terminating inhibitors, Proc. Natl. Acad. Sci. U.S.A. 74 (1977) 5463-5467]. With the completion of the human genome sequence [Venter, J.C. et al., The sequence of the human genome, Science 291 (2001) 1304-1351; Lander, E.S. et al., Initial sequencing and analysis of the human genome, Nature 409 (2001) 860-921], there is now a focus on developing new sequencing methodologies that will enable "personal genomics", or the routine study of our individual genomes. Technologies that will lead us to this lofty goal are those that can provide improvements in three areas: read length, throughput, and cost. As progress is made in this field, large sections of genomes and then whole genomes of individuals will become increasingly more facile to sequence. SNP discovery efforts will be enhanced lock step with these improvements. Here, the breadth of new sequencing approaches will be summarized including their status and prospects for enabling personal genomics. PMID- 15829236 TI - SNP discovery in associating genetic variation with human disease phenotypes. AB - With the completion of the human genome project, attention is now rapidly shifting towards the study of individual genetic variation. The most abundant source of genetic variation in the human genome is represented by single nucleotide polymorphisms (SNPs), which can account for heritable inter-individual differences in complex phenotypes. Identification of SNPs that contribute to susceptibility to common diseases will provide highly accurate diagnostic information that will facilitate early diagnosis, prevention, and treatment of human diseases. Over the past several years, the advancement of increasingly high throughput and cost-effective methods to discover and measure SNPs has begun to open the door towards this endeavor. Genetic association studies are considered to be an effective approach towards the detection of SNPs with moderate effects, as in most common diseases with complex phenotypes. This requires careful study design, analysis and interpretation. In this review, we discuss genetic association studies and address the prospect for candidate gene association studies, comparing the strengths and weaknesses of indirect and direct study designs. Our focus is on the continuous need for SNP discovery methods and the use of currently available prescreening methods for large-scale genetic epidemiological research until more advanced sequencing methods currently under development will become available. PMID- 15829237 TI - Genetic association studies of complex traits: design and analysis issues. AB - Most common diseases and many important quantitative traits are complex genetic traits, with multiple genetic and environmental variables contributing to the observed phenotype. Because of the multi-factorial nature of complex traits, each individual genetic variant generally has only a modest effect, and the interaction of genetic variants with each other or with environmental factors can potentially be quite important in determining the observed phenotype. It remains largely unknown what sort of genetic variants explain inherited variation in complex traits, but recent evidence suggests that common genetic variants will explain at least some of the inherited variation in susceptibility to common disease. Genetic association studies, in which the allele or genotype frequencies at markers are determined in affected individuals and compared with those of controls (either population- or family-based), may be an effective approach to detecting the effects of common variants with modest effects. With the explosion in single nucleotide polymorphism (SNP) discovery and genotyping technologies, large-scale association studies have become feasible, and small-scale association studies have become plentiful. We review the different types of association studies and discuss issues that are important to consider when performing and interpreting association studies of complex genetic traits. Heritable and accurately measured phenotypes, carefully matched large samples, well-chosen genetic markers, and adequate standards in genotyping, analysis, and interpretation are all integral parts of a high-quality association study. PMID- 15829238 TI - High-throughput SNP genotyping on universal bead arrays. AB - We have developed a flexible, accurate and highly multiplexed SNP genotyping assay for high-throughput genetic analysis of large populations on a bead array platform. The novel genotyping system combines high assay conversion rate and data quality with >1500 multiplexing, and Array of Arrays formats. Genotyping assay oligos corresponding to specific SNP sequences are each linked to a unique sequence (address) that can hybridize to its complementary strand on universal arrays. The arrays are made of beads located in microwells of optical fiber bundles (Sentrix Array Matrix) or silicon slides (Sentrix BeadChip). The optical fiber bundles are further organized into a matrix that matches a 96-well microtiter plate. The arrays on the silicon slides are multi-channel pipette compatible for loading multiple samples onto a single silicon slide. These formats allow many samples to be processed in parallel. This genotyping system enables investigators to generate approximately 300,000 genotypes per day with minimal equipment requirements and greater than 1.6 million genotypes per day in a robotics-assisted process. With a streamlined and comprehensive assay, this system brings a new level of flexibility, throughput, and affordability to genetic research. PMID- 15829239 TI - A single nucleotide polymorphism based approach for the identification and characterization of gene expression modulation using MassARRAY. AB - Single nucleotide polymorphisms (SNPs) are the most common form of genetic variation. Their abundance and the ease with which they can be assayed have lead to their use in applications beyond simple genotyping. One such application is the quantitative determination of transcript levels associated with distinct alleles or haplotypes found in promoters and coding regions of genes. These changes in expression due to allelic variation are often associated with additional genomic or transcript modifications such as DNA methylation or RNA editing. Here, we describe the use of an integrated genetic analysis platform, based on matrix-assisted laser desorption/ionisation-time-of-flight (MALDI-TOF) to first, discover coding SNPs (cSNPs); second, use these cSNPs to identify and analyze allele-specific expression; and third, from this knowledge to further analyze methylation patterns as a putative cause for the allele-specific expression. An established model involving allele-specific expression profiles of the human tumor protein 73 (TP73) gene is presented as an example to outline and validate data obtained from the MassARRAY platform. The availability of a single integrated platform to assay stable and dynamic variation at the genomic and transcript level greatly simplifies complex functional genomic studies. PMID- 15829240 TI - Genetic variation analyses by Pyrosequencing. AB - Pyrosequencing is a real-time bioluminometric technique for determination of nucleic acid sequence. Here, we review recent advances and discuss new applications of this technique. Cost reduction efforts and future potentials of this technique for large-scale genotyping applications will also be discussed. PMID- 15829241 TI - The Invader assay for SNP genotyping. AB - The Invader assay uses a structure-specific flap endonuclease (FEN) to cleave a three-dimensional complex formed by hybridization of allele-specific overlapping oligonucleotides to target DNA containing a single nucleotide polymorphism (SNP) site. Annealing of the oligonucleotide complementary to the SNP allele in the target molecule triggers the cleavage of the oligonucleotide by cleavase, a thermostable FEN. Cleavage can be detected by several different approaches. Most commonly, the cleavage product triggers a secondary cleavage reaction on a fluorescence resonance energy transfer (FRET) cassette to release a fluorescent signal. Alternatively, the cleavage can be detected directly by use of fluorescence polarization (FP) probes, or by mass spectrometry. The invasive cleavage reaction is highly specific, has a low failure rate, and can detect zeptomol quantities of target DNA. While the assay traditionally has been used to interrogate one SNP in one sample per reaction, novel chip- or bead-based approaches have been tested to make this efficient and accurate assay adaptable to multiplexing and high-throughput SNP genotyping. PMID- 15829242 TI - Assessment of two flexible and compatible SNP genotyping platforms: TaqMan SNP Genotyping Assays and the SNPlex Genotyping System. AB - In this review we describe the principles, protocols, and applications of two commercially available SNP genotyping platforms, the TaqMan SNP Genotyping Assays and the SNPlex Genotyping System. Combined, these two technologies meet the requirements of multiple SNP applications in genetics research and pharmacogenetics. We also describe a set of SNP selection tools and validated assay resources which we developed to accelerate the cycle of experimentation on these platforms. Criteria for selecting the more appropriate of these two genotyping technologies are presented: the genetic architecture of the trait of interest, the throughput required, and the number of SNPs and samples needed for a successful study. Overall, the TaqMan assay format is suitable for low- to mid throughput applications in which a high assay conversion rate, simple assay workflow, and low cost of automation are desirable. The SNPlex Genotyping System, on the other hand, is well suited for SNP applications in which throughput and cost-efficiency are essential, e.g., applications requiring either the testing of large numbers of SNPs and samples, or the flexibility to select various SNP subsets. PMID- 15829243 TI - Recent findings on genes associated with inflammatory disease. AB - Inflammatory diseases encompass a variety of medical conditions. In this chapter, autoimmune diseases and allergic disorders will be our focus. The autoimmune diseases include organ-specific autoimmunities, such as type I diabetes mellitus and autoimmune thyroiditis (AITD), and organ non-specific disorders such as systemic lupus erythematosus (SLE). All of them seem to share aspects of aberrant immunologic tolerance toward self-antigens. Asthma and atopic diathesis are among the allergies. Crohn disease and SLE are relatively rare with a prevalence of 10 50 per 100,000, and rheumatoid arthritis (RA), psoriasis, AITD and asthma are commoner with a prevalence of 500 per 100,000 or much higher. The difference among ethnic groups is not prominent for rheumatoid arthritis, psoriasis, AITD or asthma, but Crohn disease and SLE affect some ethnic populations more than others. Although all of these disorders have some environmental component, asthma and atopy seem most affected by environmental factors, as is suggested by the significant increase in their incidence over the last several decades with changes in various environmental factors, especially in developed countries. Over the last 10 years, multiple linkage studies revealed many disease-linked loci throughout the genome with various consistencies. As implicated by some pathophysiological studies of inflammatory immune system related disorders, certain loci are involved in multiple disorders. In the following sections, reports on the identification of disease-associated genes or markers will be summarized for individual diseases (cytotoxic T lymphocyte-associated 4 (CTLA4), CARD15, DLG5, SLC22A4/A5, programmed cell death 1 (PDCD1), RUNX1, SLC9A3R1/NAT9, PADI4, ADAM33, DPP10, PHF11 and GPRA), followed by a discussion of the genes that have been implicated in multiple disorders. PMID- 15829244 TI - Genetic association analysis: lessons from the study of Alzheimers disease. AB - Association studies are an increasingly popular way to test single nucleotide polymorphisms and other gene sequence variations for their involvement in complex disease. Alzheimers Disease has provided an ideal test-bed for the field, and it continues to be a particularly instructive disease model. Research findings demonstrate the difficulties faced by the field, and help to suggest effective ways to improve the phenotype and the genotype aspects of research practice in the field. Technical obstacles remain to be solved, but they have been well defined by early experiences with SNP and haplotype analyses, the elucidation of other confounding forms of genome variation, and the emergence of a global haplotype map. With the adoption of more sophisticated research strategies and continued advancement in high-throughput genotyping technologies, many genetic contributors to common disease will probably be identified, leading to better predictive and diagnostic medicine in the relatively near future. PMID- 15829245 TI - Mutation detection, interpretation, and applications in the clinical laboratory setting. AB - Mutation detection plays an increasingly significant role in clinical diagnostic testing, posing formidable challenges for laboratories. The expanding indications for clinical molecular testing and the nuances of interpreting test results are discussed. Methods for screening mutation detection platforms and monitoring assay reliability are presented, and results of platform comparisons are described. The potential for irrevocable medical interventions following a positive mutation analysis is highlighted to stress the imperative for accuracy in mutation detection and vigilance in the clinical arena. PMID- 15829246 TI - Single nucleotide polymorphisms in clinical genetic testing: the characterization of the clinical significance of genetic variants and their application in clinical research for BRCA1. AB - Clinical genetic testing is increasingly employed in the medical management of cancer patients. These tests support a variety of clinical decisions by providing results that indicate risk for future disease, confirmation of diagnoses, and more recently, therapeutic selection and prognosis. Most genetic variation detected during clinical testing involves single nucleotide polymorphisms (SNPs). Continued advances in the technologies of genetic analyses make these tests increasingly sensitive, cost-effective and timely, which contribute to their increased utilization. Conversely, it has proven difficult to characterize the clinical significance of genetic variants that do not obviously truncate the open reading frames of genes. These genetic variants of uncertain clinical significance diminish the value of genetic test results. This article highlights a variety of approaches that have emerged from research in diverse disciplines to solve the problem, including the application of information about common SNPs in multiple methods to better characterize clinically uncertain variants. Hereditary breast/ovarian cancer, and in particular BRCA1, provides a framework for this discussion. BRCA1 is particularly interesting in this respect since clinical genetic testing by direct DNA sequencing for over 50,000 patients in North America has revealed approximately 1500 genetic variants to date. This large data set combined with the clinical significance of BRCA1 have resulted in research groups selecting BRCA1 as a preferred gene to evaluate novel methods in this field. Finally, the lessons learned through work with BRCA1 are highly applicable to many other genes associated with cancer risk. PMID- 15829247 TI - Pharmacogenetic testing: proofs of principle and pharmacoeconomic implications. AB - Several proofs of principle have established that pharmacogenetic testing for mutations altering expression and functions of genes associated with drug disposition and response can decrease the "trial-and-error" dosing and reduce the risk of adverse drug reactions. These proofs of principle include thiopurine methyltransferase and thiopurine therapy, dihydropyrimidine dehydrogenase/thymidylate synthase and 5-fluorouracil therapy, folate enzyme MTHFR and methotrexate therapy, UGT1A1 and irinotecan therapy and CYP450 2C9 and S-warfarin therapy. These evidences advocate for the prospective identification of mutations associated with drug response, serious adverse reactions and treatment failure. More recent evidence with the HLA basis of hypersensitivity to the retroviral agent abacavir demonstrates the potential of pharmacogenetic testing and its pharmacoeconomic implications. With the convergence of rising drug costs and evidence supporting the clinical benefits of pharmacogenetic testing, it will be important to demonstrate the improved net health outcomes attributed to the additional costs for this testing. PMID- 15829248 TI - Gene SNPs and mutations in clinical genetic testing: haplotype-based testing and analysis. AB - Haplotype-based analysis using high-density single nucleotide polymorphism (SNP) markers have gained increasing attention in evaluating candidate genes in various clinical situations. For example, haplotype information is useful for predicting the severity and prognosis of certain genetic disorders. The intragenic cis interactions between the common polymorphisms and the pathogenic mutations of prion protein (PRNP) and cystic fibrosis transmembrane conductance regulator (CFTR) genes greatly influence the phenotypes and the disease penetrance of hereditary Creutzfeldt-Jakob disease and cystic fibrosis. Merits of haplotype study are more evident in the fine mapping of complex diseases and in identifying genetic variations that influence individual's response to drugs. Knowledge-based approaches and/or linkage analyses using SNP tagged haplotypes are effective tools in detecting genetic associations. For example, haplotype studies in the inflammatory bowel disease susceptibility loci revealed diverse cis and trans gene-gene interactions, which can affect the clinical outcomes. Although currently, we have very limited knowledge on haplotype-phenotypic characterizations of most genes, these examples demonstrate that increased understanding of the clinically relevant haplotypes will provide better results in the diagnosis and possibly in the treatment of both monogenic and polygenic diseases. PMID- 15829249 TI - Approaches to determine clinical significance of genetic variants. AB - The clinical significance of genetic variants (single nucleotide polymorphisms, SNPs) has implications for risk assessment and also for predicting the outcome of a disease process, especially in response to intervention. Approaches to determine the clinical significance of genetic polymorphisms are now beginning to be developed. The technology tools and procedures currently available have significant potential in identifying and validating polymorphisms associated with environmentally sensitive phenotypes. Numerous concepts can now provide the methodology to selectively identify SNPs with the potential for impacting gene function. These include computational algorithms, biochemical assays, yeast mutagenicity assays, and epidemiological studies, either as a stand-alone screen, or in various combinations depending on the gene of interest. Proof of principle will ultimately depend on large-scale epidemiological and clinical studies, but will require intensive resources. Therefore, the use of the mouse as a preclinical biological model is paramount in helping screen valid SNPs or combinations of SNPs for human studies. But more importantly, mouse modeling will help answer the question of what role gene variants play in sensitivity or resistance to a wide variety of environmental insults ranging from toxic chemicals and carcinogens to more mundane and routine exposure items, such as dietary factors, air quality, over the counter and prescription medications, and ultraviolet light. Our focus on SNPs that result in an amino acid change is a matter of expediency because these variants are more amenable to the prescreening approaches currently available that are expected to help identify SNPs that affect protein function. The mouse models generated to evaluate the environmental relevance of selected SNPs will be extremely valuable biological tools to validate gene variant and environment interaction in a variety of settings. Informative mouse models will also provide the basis of pursuing relevant SNPs in epidemiological and clinical investigations. PMID- 15829250 TI - Behavioral and convulsant effects of the (S) enantiomer of the group I metabotropic glutamate receptor agonist 3,5-DHPG in mice. AB - The purpose of the present studies was to investigate the behavioral and convulsant effects produced by the group I metabotropic glutamate receptor agonist (S)-3,5-dihydroxyphenylglycine (DHPG). Administered i.c.v. to mice, (S) 3,5-DHPG produced a behavioral syndrome consisting of scratching and/or facial grooming, tremors, slow forelimb clonus, rearing, and falling that increased over the dose range of 10-400 nmol. The full syndrome, produced by 400 nmol of (S)-3,5 DHPG, was antagonized by the selective mGlu1 receptor antagonist LY456236 but not by the mGlu5 receptor antagonist MPEP or the mGlu2/3 receptor antagonist LY341495. The behaviors induced by the 400 nmol dose were not blocked by the NMDA receptor antagonist MK-801, but were attenuated by the non-NMDA receptor antagonists GYKI 52466 and NBQX, and the Ca2+ mobilization inhibitor dantrolene, but at motor-impairing doses. The scratching behaviors produced by 30 nmol of (S) 3,5-DHPG were antagonized by LY456236 but not by MPEP, LY341495 or MK-801. GYKI 52466 and dantrolene, but not NBQX, inhibited scratching at motor-impairing doses. Both 400 and 30 nmol of (S)-3,5-DHPG produced a generalized seizure as recorded by surface EEG electrodes. LY456236 blocked the seizures produced by 30 nmol but not by 400 nmol; dantrolene was ineffective in blocking seizures produced by either dose. The present findings suggest that (S)-3,5-DHPG produces an increase in excitation that is mediated by mGlu1 and non-NMDA receptors. PMID- 15829251 TI - Kynurenine 3-mono-oxygenase inhibitors reduce glutamate concentration in the extracellular spaces of the basal ganglia but not in those of the cortex or hippocampus. AB - Kynurenine 3-mono-oxygenase (KMO, kynurenine hydroxylase) inhibitors increase brain kynurenic acid (KYNA) synthesis and cause pharmacological actions possibly mediated by a reduced activity of excitatory synapses. We used in vivo microdialysis and passive avoidance to study the effects of local KYNA or systemic KMO inhibitor administration on glutamate (GLU) neurotransmission. Local application of KYNA (30-100 nM) through reverse microdialysis reduced GLU content in caudate and cortical dialysates by 75 and 55%, respectively. No changes were found in the hippocampus. Systemic administration of Ro 61-8048 (4-40 mg/kg) increased KYNA levels in dialysates obtained from the cortex (from 10.3 +/- 1.9 to 45.5 +/- 15 nM), caudate (from 2.4 +/- 0.8 to 9.5 +/- 0.9 nM) and hippocampus (from 7.7 +/- 1.7 to 19.2 +/- 3.5 nM). It also caused a parallel robust decrease in GLU levels in the dialysates collected from the caudate (from 2.2 +/- 0.5 to 0.63 +/- 0.05 microM) but not in those collected from the parietal cortex or the hippocampus. In a passive avoidance paradigm, the administration of the NMDA receptor antagonist MK-801 (0.1 mg/kg) reduced, while Ro 61-8048 (4-80 mg/kg) did not change the latency time of entering into the dark compartment on the recall trial. Our data show that KMO inhibitors increase brain KYNA synthesis and selectively reduce GLU extracellular concentration in the basal ganglia. PMID- 15829252 TI - M3-like muscarinic receptors mediate Ca2+ influx in rat mesencephalic GABAergic neurones through a protein kinase C-dependent mechanism. AB - GABAergic neurones in the mesencephalon are important regulators of dopamine neurones. Cholinergic projections from mesopontine nuclei preferentially synapse onto these GABAergic neurones, thus suggesting that ACh can regulate dopamine neurones indirectly by modulating GABAergic interneurones. Muscarinic receptors mediate excitation of these interneurones through a Ca(2+)-dependent mechanism. Using a mesencephalic primary culture model, we show here that muscarine (10 microM) increases intracellular Ca2+ concentrations ([Ca2+]i) in GABAergic interneurones. Compatible with previous anatomical data, our pharmacological studies further suggest that the M3 receptor is the primary mediator of this increase. The rise in [Ca2+]i induced by muscarine was not activity-dependent but required influx of Ca2+ from the extracellular medium. Consistent with the known coupling of the M3 receptor to PKC, the effect of muscarine was blocked by bisindolylmaleimide, a selective PKC antagonist. The effect of muscarine was inhibited by SKF 96365 and verapamil, drugs known to block non-selective cationic channels such as those formed by transient receptor potential (TRPC) proteins. Finally, GABAergic neurones were found to be immunopositive for TRPC1, 3, 5 and 6. Taken together, these results suggest that the Ca(2+)-dependent regulation of mesencephalic GABAergic neurones by muscarinic receptors requires activation of some receptor-operated Ca2+ channels through a PKC-dependent mechanism. PMID- 15829253 TI - In vivo treatment with the K+ channel blocker 4-aminopyridine protects against kainate-induced neuronal cell death through activation of NMDA receptors in murine hippocampus. AB - Activation of NMDA receptors has been shown to induce either neuronal cell death or neuroprotection against excitotoxicity in cultured neurons in vitro. To elucidate in vivo neuroprotective role of NMDA receptors, we investigated the effects of activation of NMDA receptors by endogenous glutamate on kainate induced neuronal damage to the mouse hippocampus in vivo. The systemic administration of the K+ channel blocker 4-aminopyridine (4-AP, 5 mg/kg, i.p.) induced expression of c-Fos in the hippocampal neuronal cell layer, which expression was completely abolished by the noncompetitive NMDA receptor antagonist MK-801, thus indicating that the administration of 4-AP would activate NMDA receptors in the hippocampal neurons. The prior administration of 4-AP at 1 h to 1 day before significantly prevented kainate-induced pyramidal cell death in the hippocampus and expression of pyramidal cells immunoreactive with an antibody against single-stranded DNA. Further immunohistochemical study on deoxyribonuclease II revealed that the pretreatment with 4-AP led to complete abolition of deoxyribonuclease II expression induced by kainate in the CA1 and CA3 pyramidal cells. The neuroprotection mediated by 4-AP was blocked by MK-801 and by the adenosine A1 antagonist 8-cyclopenthyltheophylline. Taken together, in vivo activation of NMDA receptors is capable of protecting against kainate induced neuronal damage through blockade of DNA fragmentation induced by deoxyribonuclease II in the murine hippocampus. PMID- 15829254 TI - The role of the NMDA receptor in alcohol relapse: a pharmacological mapping study using the alcohol deprivation effect. AB - Modulators of glutamate receptors especially of the N-methyl-D-aspartate receptors (NMDARs) have recently been suggested as putative pharmacotherapeutic agents in the treatment of alcohol relapse. However, at present it is not clear, which binding and modulatory sites of the NMDAR are involved in relapse behavior. We, therefore, performed a pharmacological mapping study in long-term alcohol drinking rats using the alcohol deprivation effect (ADE) as a model for relapse behavior. In a comprehensive fashion, we studied dose-response curves, employing the following selective pharmacological agents: the NMDAR competitive antagonist CGP37849, the glycine binding site antagonist L-701.324, the NR2B subunit selective antagonist ifenprodil, which acts at the polyamine binding site, the NMDAR channel blocker neramexane, and ethanol, which acts as a functional antagonist at the NMDAR. Our data show that the animals' alcohol consumption inversely correlates with the dose of ethanol administered intraperitoneally. This indicates that under the present experimental conditions alcohol intake during an ADE is an entirely pharmacologically driven behavior that is not under the control of other factors such as taste or novelty of alcohol re-exposure. The effects of the administration of the aforementioned compounds were comparable to those of ethanol, suggesting a similar pharmacological impact on relapse behavior. Repeated administration of both competitive and uncompetitive NMDAR antagonist dose-dependently suppressed alcohol consumption during ADE. In addition, ifenprodil and L-701.324 dose-dependently reduced the expression of an ADE as well. In summary, the results suggest that an inhibition of NMDAR function in general, rather than a particular interference with a specific binding site of this receptor, is sufficient for the reduction of relapse behavior. PMID- 15829255 TI - Analysis of the role of 5-HT1A receptors in spatial and aversive learning in the rat. AB - The role of the brain 5-HT1A receptor in cognition was examined in the water maze (WM) and passive avoidance (PA) tasks in the male rat. Pre-training administration of the 5-HT1A receptor agonist 8-OH-DPAT impaired WM performance and facilitated PA retention at low doses (0.01 and 0.03 mg/kg) and impaired PA retention at higher doses (0.1-1.0 mg/kg). The 5-HT1A receptor antagonist NAD-299 produced a dose-dependent facilitation of PA retention. In contrast, the 5-HT1A receptor antagonists NAD-299 and WAY-100635 failed to alter acquisition and retention in the WM. The impairments in WM and PA (but not facilitation in PA) induced by 8-OH-DPAT were blocked by NAD-299. Furthermore, NAD-299 prevented the PA impairments induced by the muscarinic antagonist scopolamine or the NMDA receptor antagonist MK-801. In contrast, NAD-299 and WAY-100635 failed to attenuate the WM impairment induced by scopolamine, probably due to the failure of 5-HT1A receptor blockade to attenuate the sensorimotor disturbances induced by scopolamine. These results indicate that 5-HT1A receptor stimulation and blockade result in opposite effects in two types of cognitive tasks in the rat, and that 5 HT1A receptor blockade can facilitate some aspects of cognitive function, probably via modulation of cholinergic and glutamatergic transmissions. This suggests that 5-HT1A receptor antagonists may have a potential role in the treatment of human degenerative disorders associated with cognitive deficits. PMID- 15829256 TI - Morphine alters the selective association between mu-opioid receptors and specific RGS proteins in mouse periaqueductal gray matter. AB - In the CNS, several regulators of G-protein signalling (RGS) modulate the activity of mu-opioid receptors. In pull-down assays performed on membranes from mouse periaqueductal gray matter (PAG), mu-opioid receptors co-precipitated with delta-opioid receptors, Gi/o/z/q proteins, and the regulators of G-protein signalling RGS4, RGS9-2, RGS14, RGSZ1 and RGSZ2. No RGS2, RGS7, RGS10 and RGS11 proteins were associated with the mu receptors in these PAG membranes. In mice, an intracerebroventricular dose of 10 nmol morphine produced acute tolerance at mu receptors but did not disrupt the co-precipitation of mu-delta receptor complexes. However, this opioid reduced by more than 50% the co-precipitation of G alpha i/o/z subunits with mu receptors, and altered their association with some of the RGS proteins at 30 min, 3 h and 24 h after its administration. The association of RGS9-2 with mu receptors diminished by 30-40% 24 h after the administration of morphine, while that of RGSZ2 and of RGSZ1 increased. Morphine treatment recruited RGS4 to the PAG membranes, and 30 min and 3 h after the opioid challenge its association with mu receptors had increased. However, 24 h after morphine administration, the co-precipitation of RGS4 had decreased by about 30%. The opioid produced no change in the membrane levels of RGS9-2, RGS14, RGSZ1 and RGSZ2. Thus, in PAG synaptosomal membranes, a dynamic and selective link exists between, mu-opioid receptors, Gi/o/z proteins and certain RGS proteins. PMID- 15829257 TI - Role of alpha7-nicotinic acetylcholine receptors in tetanic stimulation-induced gamma oscillations in rat hippocampal slices. AB - Hippocampal gamma oscillations, as a form of neuronal network synchronization, are speculated to be associated with learning, memory and attention. Nicotinic acetylcholine receptor alpha7 subtypes (alpha7-nAChRs) are highly expressed in hippocampal neurons and play important roles in modulating neuronal function, synaptic plasticity, learning and memory. However, little is known about the role of alpha7-nAChRs in hippocampal gamma oscillations. Here, we examined the effects of selective alpha7- and non-alpha7-nAChR antagonists on tetanic gamma oscillations in rat hippocampal slices. We found that brief tetanic stimulation induced gamma oscillations (30-80 Hz) and pharmacological blockade of alpha7 nAChRs using the relatively selective alpha7-nAChR antagonists, methyllycaconitine (10 or 100 nM) or alpha-bungarotoxin (10 nM), significantly reduced the frequency spectrum power, the number of spikes, and burst duration of evoked gamma oscillations. Neither mecamylamine nor dihydro-beta-erythroidine, which are selective antagonists of non-alpha7-nAChRs, demonstrated significant effects on tetanic gamma oscillations. Nicotine exposure promotes hippocampal gamma oscillations in a methyllycaconitine-sensitive manner. It is concluded that alpha7-nAChRs in hippocampal slices play important roles in regulation of gamma oscillations, thus potentially helping to explain roles of nAChRs in cognitive functions such as learning, memory and attention. PMID- 15829258 TI - Binding and functional properties of the novel somatostatin analogue KE 108 at native mouse somatostatin receptors. AB - Clinically used somatostatin (SRIF) analogs, octreotide and lanreotide, act primarily by binding to SRIF receptor subtype 2 (sst2). In contrast, the recently described multiligand SOM230 binds with high affinity to sst(1-3) and sst5 and KE 108 is characterised as a high affinity ligand for all five SRIF receptors. In tumoural mouse corticotrophs (AtT-20 cells) and in mouse hippocampus, binding and functional features of KE 108 were examined and compared to SRIF-14, octreotide and SOM230. In AtT-20 cells, KE 108 bound with high affinity at [125I]LTT-SRIF-28 labelled sites similarly to SRIF-14, octreotide and SOM230. At the functional level, all four ligands increased guanosine-5'-O-(3-[35S]thio)-triphosphate binding and decreased cAMP accumulation or intracellular Ca2+ concentration through G(i/o) proteins. In hippocampal slices, KE 108, octreotide and SOM230 also bound with high affinity at [125I]LTT-SRIF-28-labelled sites similarly to SRIF-14, but KE 108, octreotide or SOM230 did not influence spontaneous epileptiform activity which was, in contrast, inhibited by SRIF-14. In conclusion, this study demonstrates that KE 108 has high affinity for native mouse SRIF receptors. Functionally, KE 108 mediates SRIF action at sst(2/5) in corticotrophs whereas it does not mimic the SRIF-induced inhibition of hippocampal excitation suggesting that the high potency and efficacy of a synthetic ligand to all known SRIF receptors may not reproduce entirely the effects of the natural SRIF. PMID- 15829259 TI - Pentylenetetrazole-induced convulsions affect cellular and molecular parameters of the mechanism of action of triiodothyronine in adult rat brain. AB - The aim of the current study was to elucidate whether the response of the adult rat brain to thyroid hormones is affected by the intensity of neuronal activity. For this purpose, the kinetic characteristics of nuclear T3 binding, the relative expression of thyroid hormone receptor (TR) isoforms and the synaptosomal content of thyroid hormones in adult rat brain were examined after administration of a single convulsion dose of pentylenetetrazole (PTZ). Experiments in adult Wistar rats revealed an increase (33%) of the density of specific T3 nuclear receptors in cerebral hemispheres 4h after PTZ-induced seizures while no changes were observed in the dissociation constant. The relative expression of the T3-binding isoforms of TRs was not affected, while there was a gradual decrease of the relative expression of the TR alpha2 variant (non-T3 binding isoform). The above changes were coupled with an increase of the synaptosomal T3 levels during the epileptic seizures. Our study revealed inversely proportional changes between the nuclear T3 binding sites and the TR alpha2 mRNA levels 4 h after PTZ-induced seizures, suggesting that the regulation of the expression of the non-T3 binding variant of TRs determines the nuclear T3 binding sites in adult rat brain, while the synaptosomal T3 levels could play a novel functional role in the signaling from the synapse to the nucleus. PMID- 15829260 TI - Specific neurodevelopmental damage in mice offspring following maternal inflammation during pregnancy. AB - Intrauterine inflammation is a major risk for offspring neurodevelopmental brain damage and may result in cognitive limitations and poor cognitive and perceptual outcomes. Pro-inflammatory cytokines, stimulated during inflammatory response, have a pleotrophic effect on neurons and glia cells. They act in a dose-dependent manner, activate cell-death pathways and also act as trophic factors. In the present study, we have examined in mice the effect of short, systemic maternal inflammation on fetal brain development. Maternal inflammation, induced by lipopolysaccharide (LPS) at gestation day 17, did not affect morphogenic parameters and reflex development during the first month of life. However, maternal inflammation specifically increased the number of pyramidal and granular cells in the hippocampus, as well as the shrinkage of pyramidal cells, but not of the granular cells. No additional major morphological differences were observed in the cerebral cortex or cerebellum. In accordance with the morphological effects, maternal inflammation specifically impaired distinct forms of learning and memory, but not motor function or exploration in the adult offspring. The specific deficiency observed, following maternal inflammation, may suggest particular sensitivity of the hippocampus and other associated brain regions to inflammatory factors during late embryonic development. PMID- 15829261 TI - Diclofenac, a nonsteroidal anti-inflammatory drug, activates the transient outward K+ current in rat cerebellar granule cells. AB - Diclofenac, a nonsteroidal anti-inflammatory drug (NSAID), has been widely investigated in terms of its pharmacological action, but less is known about its direct effect on ion channels. Here, the effect of diclofenac on voltage dependent transient outward K+ currents (I(A)) in cultured rat cerebellar granule cells was investigated using the whole-cell voltage-clamp technique. At concentrations of 10(-5)-10(-3) M, diclofenac reversibly increased the I(A) amplitude in a dose-dependent manner and significantly modulated the steady-state inactivation properties of the I(A) channels, but did not alter the steady-state activation properties. Furthermore, diclofenac treatment resulted in a slightly accelerated recovery from I(A) channel inactivation. Intracellular application of diclofenac could mimic the effects induced by extracellular application, although once the intracellular response reached a plateau, extracellular application of diclofenac could induce further increases in the current. These observations indicate that diclofenac might exert its effects on the channel protein at both the inner and outer sides of the cell membrane. Our data provide the first evidence that diclofenac is able to activate transient outward potassium channels in neurons. Although further work will be necessary to define the exact mechanism of diclofenac-induced I(A) channel activation, this study provides evidence that the nonsteroidal anti-inflammatory drug, diclofenac, may play a novel neuronal role that is worthy of future study. PMID- 15829262 TI - Proceedings of the 2nd International Conference "B Cells and Autoimmunity: New Concepts and Therapeutic Perspectives". Quebec City, Canada, July 15-18, 2004. PMID- 15829263 TI - Regulatory events in early and late B-cell differentiation. AB - We are studying transcriptional control of critical developmental decision points in B lymphocytes. Commitment to the B-lymphocyte lineage is dependent on the transcriptional regulator Pax5 and committed B lymphocytes represent the first developmental stage when V(H)-to-DJ recombination occurs in the immunoglobulin (Ig) heavy chain locus. We summarize our recent studies showing that methylation of histone H3 lysine 9, a heterochromatic chromatin modification, is present in the Ig V(H) region in hematopoietic progenitors and in non-B lineage hematopoietic cells. Pax5 is both necessary and sufficient to remove this heterochromatic mark in B cells. Using genetically altered mice, we have shown that terminal differentiation of B cells to memory and Ig-secreting plasma cells depends on the transcriptional repressor Blimp-1. Recent studies demonstrating a requirement for Blimp-1 in the formation of pre-plasma memory B cells, Ig secreting plasma cells as well as preliminary data suggesting a requirement for Blimp-1 in the maintenance of long-lived plasma cells are summarized. We also summarize our recent studies on the regulation of Blimp-1, showing direct repression by Bcl-6 and providing evidence for activation by NF-kappaB following toll-like receptor signaling. PMID- 15829264 TI - The BAFF/APRIL system: life beyond B lymphocytes. AB - B cell activating factor belonging to the TNF family (BAFF) and a proliferation inducing ligand (APRIL) are two members of the TNF ligand superfamily. Studies of BAFF, APRIL and their receptors have highlighted the importance of this ligand/receptor system in regulating B cell homeostasis, tolerance and malignancy. Neutralizing BAFF can inhibit disease progression in animal models of autoimmunity, possibly by reducing survival of autoreactive B cells. In addition, BAFF inhibitors also prevent B lymphoma cell survival and may be useful for the treatment of lymphoid cancers. Recent work suggests that BAFF is also important for T cell activation and differentiation, an aspect that may be critical for the progression of certain autoimmune diseases. Therefore, targeting the BAFF/APRIL system may protect against autoimmunity and lymphoid cancers through the inhibition of pathogenic B and T cell functions. PMID- 15829265 TI - Requirement of the extracellular cysteine at position six for CD40/CD40 dimer formation and CD40-induced IL-8 expression. AB - We recently showed that oligomerization of CD40 molecules on cell surface leads to disulfide-linked CD40/CD40 dimer formation, an event that is necessary for CD40-induced B7-2 expression in human B cells. Here, we demonstrate that CD40/CD40 dimers formation also occurs in different cell types such as T24 bladder cancer cells and CD40-transfected HEK 293 cells. Disulfide bonds mediate the formation of CD40/CD40 homodimers in CD40-activated cells. To determine the potential residue(s) involved in disulfide bonds formation and subsequent CD40 induced IL-8 expression, we generated a CD40 mutant in which the extracellular cysteine 6 was replaced by a glutamine (CD40-C6Q). CD40-induced IL-8 mRNA expression and protein synthesis were studied in stably transfected HEK 293 cells that were sorted out along with similar levels of expression of wild type (CD40 WT) and CD40-C6Q molecules. In contrast to cells expressing CD40-WT protein, disulfide-linked CD40/CD40 dimer formation was completely abolished in HEK 293 cells expressing CD40-C6Q proteins. Abolishment of disulfide-linked CD40/CD40 dimers in these transfected cells was sufficient to inhibit CD40-induced mRNA expression and secretion of IL-8. This study identifies the extracellular cysteine 6 of CD40 molecules as a potential molecular target to disrupt the expression of CD40-induced pro-inflammatory cytokines by epithelial cells. PMID- 15829266 TI - The role of IgM antibodies in the recognition and clearance of apoptotic cells. AB - Apoptotic cells not only translocate phosphatidylserine to the outside of the membrane, they activate phospholipases that remodel their membranes resulting in the exposure and diffusion of altered phospholipids. These altered phospholipids include lysophosphotidylcholine (LPC) that is a ligand for CRP and is also antigenic for natural IgM antibodies. Furthermore, soluble LPC acts as a "find me" signal to attract phagocytes to migrate toward apoptotic cells. IgM mediated opsonization of apoptotic cells with complement accelerates their clearance by phagocytic cells. Failure to rapidly remove apoptotic cells on the other hand, allows dying cells to undergo post-apoptotic necrosis and inflammation. Natural antibodies therefore provide a previously unrecognized role in prevention of autoimmunity. PMID- 15829267 TI - Adult lupus-prone MRL/MpJ2+ mice express a primary antibody repertoire that differs in CDR-H3 length distribution and hydrophobicity from that expressed in the C3H parental strain. AB - Anti-dsDNA antibodies tend to be enriched for heavy chain complementarity determining region 3 (CDR-H3) intervals of above average length that contain an increased frequency of charged amino acids. It is unclear whether these types of CDR-H3s are more common in the primary B-cell repertoire of auto-immune prone strains or whether their increased prevalence in affected individuals reflects positive selection and expansion of atypical CDR-H3s in the pathogenic response to self-antigen. Here, we present evidence that when compared to C3H, a MRL/MpJ(2+) parental strain, CDR-H3 intervals from pre-B cells of adult lupus prone MRL/MpJ(2+) mice are longer on average and are enriched for charged amino acids. The predicted prevalence of deformed loops per Shirai H3 criteria is also higher. In contrast, the frequency of charge, the distribution of length, and the pattern of predicted deformed loop structures did not differ in sequences obtained from neonates of the same two strains. These observations suggest that the mechanisms that serve to shape the initial CDR-H3 repertoire in adults, but not neonates, are being regulated differently in C3H versus MRL/MpJ(2+). Dysregulation of the adult pre-B CDR-H3 antibody repertoire could be a contributing factor for the development of florid auto-immune disease in MRL/MpJ(2+) mice. PMID- 15829268 TI - Chemokine-mediated control of T cell traffic in lymphoid and peripheral tissues. AB - Antigen-driven T cell education and subsequent pathogen elimination present particular challenges for the immune system. Pathogens generally enter the body at peripheral sites such as the skin, gastrointestinal tract or lung, areas from which naive T cells are largely excluded. Instead, naive T cells constantly recirculate through secondary lymphoid organs, such as lymph nodes and Peyer's patches, in search for antigen brought to these locations by means of afferent lymphatic channels. Here, antigen-loaded dendritic cells present antigen-peptide MHC complexes to clonotypic T cells and provide appropriate co-stimulatory signals for immune response initiation. As a result, short-lived effector T cells and long-lived memory T cells are generated that reach the peripheral tissue for participation in immune responses and immune surveillance. Effector and memory T cell relocation is non-random, due to tissue-specific "address codes" that allow proper tissue homing. This process involves adhesion molecules, including selectins, integrins, and corresponding vascular ligands as well as the large family of chemokines and their receptors. Here, we discuss the changes in chemokine receptor expression that occur during T cell activation and differentiation, and the ways in which these changes impact on the migration potential of naive, effector, and memory T cells. We summarize our current understanding of T cell homing to the T zone and B cell follicles within secondary lymphoid tissues and highlight the two chemokine receptors CCR7 and CXCR5 that recognize chemokines constitutively present either in the T zone (CCR7 ligands CCL19/ELC and CCL21/SLC) or follicular compartment (CXCR5 ligand CXCL13/BCA-1). CCR7 is characteristic for naive and central memory T (T(CM)) cells whereas CXCR5 distinguishes follicular B helper T (T(FH)) cells. In addition, we further subdivide long-lived memory T cells into CCR7-negative effector memory T (T(EM)) cells and peripheral immune surveillance T (T(PS)) cells. The latter term designates the extraordinarily large subset of memory T cells with primary residence in normal (healthy) peripheral tissues. Our current understanding of T(PS) cell migration and function is highly fragmentary, but these cells are thought to provide immediate protection locally at the site of pathogen entry. Here, we propose that the tissue distribution of T(PS) cells is determined by a distinct set of chemokines and corresponding receptors that differs from those operating in secondary lymphoid tissues and inflammatory sites. PMID- 15829269 TI - Hormonal modulation of B cell development and repertoire selection. AB - Systemic lupus erythematosus is an autoimmune disorder characterized by the production of pathogenic autoantibodies, primarily to nuclear antigens. The etiology of SLE is not entirely understood, but it is well-appreciated that multiple factors such as genetics and environment contribute to disease progression and pathogenesis. There is also convincing evidence that gender plays an import role in SLE since the incidence of disease occurs with a female to male ratio of 9:1. While it is plausible that some sex-linked genes may contribute to the genetic predisposition for the disease, other likely culprits for this gender bias are the sex hormones estrogen and prolactin. The data implicating estrogen and prolactin in SLE, until recently, were largely circumstantial. However, within the last few years, data collected from both human and mouse studies have provided compelling evidence that alterations in sex hormone levels can alter tolerance of autoreactive B cells and exacerbate disease. In this review, we will discuss recent data demonstrating a role for estrogen and prolactin in SLE and the effect of these hormones on B cell maturation, selection and activation. PMID- 15829271 TI - Mechanisms of heavy metal-induced autoimmunity. AB - Chemical exposure can trigger or accelerate the development of autoimmune manifestations. Although heavy metals are elementary chemical structures, they can have profound and complex effects on the immune system. In genetically susceptible mice or rats, administration of subtoxic doses of mercury induces both the production of highly specific autoantibodies and a polyclonal activation of the immune system. We review in this article some of the mechanisms by which heavy metal exposure can lead to autoimmunity. PMID- 15829272 TI - Therapeutic intravenous immunoglobulins. AB - Intravenous immunoglobulins (IVIg) are concentrated formulations of human IgG prepared by industrial fractionation of large pools of individual plasma donations. IVIg were developed 20 years ago for the prophylaxis support of immunodeficient patients. However, IVIg have been increasingly used since 10 years, in the treatment of many autoimmune and inflammatory diseases raising the possibility of product shortages and ever increasing costs in the near future. Surprisingly, the immunomodulatory mechanisms of action of IVIg are unclear because of the diversity and often contradictory Fc, F(ab')(2), and non-IgG related mechanisms that have been proposed from clinical observations and from results obtained in various in vitro and in vivo experimental models. These concepts are reviewed here and we discuss in more details three areas of active research, namely the mechanisms of IVIg action in Idiopathic Thrombocytopenic Purpura (ITP), the effects of IVIg on activated B lymphocytes and the possible involvement of autoantibodies of IgG isotype (auto-IgG) in the immunomodulatory effects of IVIg. The elucidation of the mechanisms of action of IVIg is crucial for a more rationalized clinical use of IVIg and for developing substitutes for some of the immunomodulatory indications in order to ensure long-term availability of plasma-derived IVIg for immunodeficient patients. PMID- 15829273 TI - Effect of the B cell superantigen protein A from S. aureus on the early lupus disease of (NZBxNZW) F1 mice. AB - The (NZBxNZW) F(1) mouse develops a spontaneous autoimmune disease process with striking similarities to human systemic lupus erythematosus (SLE). In female (NZBxNZW) F(1) mice, the production of IgG antinuclear antibodies, including antibodies to double-stranded DNA (dsDNA), is associated with the development of a severe immune complex-mediated glomerulonephritis that results in death from renal failure in virtually all animals by 12 months of age. Since B-1 and marginal zone (MZ) cells represent a potential source of pathogenic antibodies and because B cell superantigens have been demonstrated to reduce B-1 and MZ cells in vivo, we tested the effect of repeated injections of the superantigen protein A (SpA) from S. aureus on the disease of this lupus model. We found that weekly intraperitoneal injections of SpA delay the progression of serum anti-DNA IgG and reduce proteinuria early in young female (NZBxNZW) F(1) mice. This superantigen also induced a specific depression in the numbers of peritoneal B-1 cells, as compared to mice treated with a control protein. These results support the role of B-1 cells in the development of the autoimmune disease in this mouse model and suggest that B cell superantigens may be useful in the management of autoimmune conditions. PMID- 15829274 TI - Innate immunity in health and disease. PMID- 15829275 TI - The role of Toll-like receptors in the host response to viruses. AB - The discovery of Toll-like receptors (TLR) has revolutionised our understanding of innate immunity. Numerous reviews have been written on the subject in the past few years. Here, we review the evidence that TLRs are involved in sensing and initiating anti-viral responses. There are now three strong lines of evidence that support such a role for TLRs. Firstly, TLRs 'recognise' virally derived molecules and are required for various virus-induced cellular effects. Secondly, TLRs trigger anti-viral signalling pathways leading to the induction of the interferon response. Thirdly, viral immune strategies employed against TLRs have been identified. PMID- 15829276 TI - Type I interferons and the innate immune response--more than just antiviral cytokines. AB - The role of type I interferon (referred to as IFN in this review) in early antiviral immunity is well known. More recently IFN has been shown to be a potent regulator of adaptive immunity. It is now becoming clear that a broad range of viruses, bacteria and even parasites express ligands capable of stimulating a growing number of signalling pathways that results in, often subtype specific, induction of IFN. Of particular interest are the signalling pathways associated with the Toll-like receptors. This family of receptors, each able to induce signals in response to a variety of ligands, initiates the pro-inflammatory response. They also contain members that have the capacity to induce IFN, making use of, and perhaps promoting the evolution of its pleiotropic responses. Greater knowledge of the events that result in induction of IFN is necessary in understanding the specificity of expression of an increasingly complex and important aspect of our immune system. This may reveal to us further therapeutic opportunities, either in the use of IFN or in the manipulation of their expression. This review details the established knowledge and recent advances made in understanding how and under what circumstances the IFNs are expressed, starting with brief overviews of IFN and Toll-like receptors before following the molecular processes from induction of IFN, activation of the JAK-STAT pathway and finally the expression of interferon stimulated genes and their functions. PMID- 15829277 TI - Innate immune recognition of the extracellular mucosal pathogen, Helicobacter pylori. AB - Toll-like receptor (TLR) molecules play a frontline role in the defence of the host against infection by microbial pathogens. These molecules, together with the recently described Nod family proteins, have been shown to trigger innate immune responses in host cells via the recognition of highly conserved microbial structures. TLR4, which is the best-characterised of these "pathogen-recognition molecules" (PRMs), was the first to be shown to recognise a specific microbial component: the lipopolysaccharide (LPS) from Gram-negative bacteria. The molecular specificities of the remaining PRMs have, in nearly all cases, now also been elucidated. Host cells belonging to the myeloid cell lineage are known to be particularly responsive to these microbial constituents. Conversely, other cell types such as epithelial cells, were generally thought to be hypo-responsive to stimulation by such molecules. New evidence suggests that these cells are in fact likely to play a fundamental role in host defence against pathogenic micro organisms. Indeed, epithelial cells afford an initial barrier against the host microflora, and appear to be able to differentiate between pathogenic and commensal micro-organisms. This review article will discuss current knowledge regarding innate immune responses in epithelial and myeloid cells to the model non-invasive pathogen, Helicobacter pylori, which is a major cause of upper gastrointestinal tract disease in humans. PMID- 15829278 TI - Intestinal epithelial TOLLerance versus inTOLLerance of commensals. AB - This brief review summarizes the current understanding of Toll-like receptor (TLRs) mediated intestinal epithelial mechanisms of commensal tolerance versus intolerance and provides an update on the downstream negative control of signaling responses through decreased surface expression, interregulation with NOD2, overexpression of Tollip, various inhibitors of NF-kappaB as well as soluble tolerizing mediators present in lumen and serum which all may maintain or -when dysregulated--impair mucosal homeostasis in health or disease, respectively. PMID- 15829279 TI - Bacterial modulation of mucosal innate immunity. AB - The human gut harbours a diverse population of non-pathogenic, commensal bacteria whose contribution to gastrointestinal health and disease is now recognised. This microflora plays an important role in the development and expansion of lymphoid tissues and in the maintenance and regulation of gut immunity. A critical feature of the mucosal immune system is the ability to discriminate between harmful pathogens and the harmless members of the commensal flora. This is achieved in part, by an evolutionary-conserved family of cell surface and cytosolic receptors, referred to as toll-like receptors (TLRs), which function in microbial recognition. Appropriate activation of TLRs has been demonstrated as an essential component of host immunity against pathogens but is also vital for immune homeostasis. The ability of TLRs to discriminate between pathogens and commensals is not clear cut, however, and hence complex regulatory systems, derived both from host and bacterial origin, appear to reinforce and support this system. Host factors that modulate and alter TLR-mediated signaling have recently been defined and are thought to control the level of immune activation. Similarly, certain gut bacteria are also recognised to suppress unnecessary inflammatory responses, thereby helping to maintain immune homeostasis. Their relative contribution to these regulatory processes is currently unknown. The host transcription factor, nuclear factor kappa B (NF-kappaB) has been consistently identified as an important target molecule for bacterial regulation. NF-kappaB, which is also essential for immune activation, is an important therapeutic target for the treatment of inflammatory bowel diseases. Hence, the possibility exists that bacterially derived effector molecules, with defined modes of action, may have clinical relevance and application. PMID- 15829280 TI - Innate immune defence in the human gastrointestinal tract. AB - The mucosal surface of the gastrointestinal tract represents a major entry point and ecological niche for many microbes. It forms an important immune barrier, absorbing nutrients, whilst preventing invasion by organisms. Of the extra ordinarily diverse species that comprise the microbial world, relatively few organisms are able to succeed in breaching this barrier in an otherwise healthy host. The production and secretion of antimicrobial peptides (AMPs) from surface epithelia and circulating immune cells are likely to play a key role in host protection and homeostasis. A number of these peptides are constitutively produced providing resident protection, whereas others are induced during infection and inflammation. In addition to directly eradicating microorganisms, it is becoming increasingly apparent that AMPs are multi-functional with diverse immuno-modulatory properties. This review focuses on three families of AMPs, defensins, cathelicidins, and lysozyme, and discusses their role in mucosal defence. PMID- 15829281 TI - Gliadin as a stimulator of innate responses in celiac disease. AB - In celiac disease (CD) we have the prototype of an immune mediated response dominated by the activation of the adaptive immune system and in particular of CD4+ HLA class II restricted T cells. Various seminal studies have established the precise mechanism of how antigen (prolamine) specific activation of CD4+ mucosal T cells occurs. Thus, CD is a condition in which T cells and their activation is the essential hinge in the pathogenic process. These functional studies have provided the explanation for the genetic association between CD and certain HLA alleles (HLA DQ2 and DQ8). These genetic, molecular and functional studies have permitted the clarification of a powerful Th1 dominated pro inflammatory response that characterises the small intestine of active CD patients. Despite this unassailable set of information and reports there are some intriguing points that have been raised by a series of studies which have indicated that CD is not only defined by an aberrant prolamine-induced activation of the adaptive immune system. New evidence and re-assessments of old studies, point to a more complex pathogenic cascade, which may help to unravel some of the residual obscure points of CD pathogenesis. Here, we outline the current concepts that indicate a direct involvement of the adaptive immune system and we discuss all the evidence supporting a direct activation of the innate immune system by fragments of prolamines, which are not recognized T cell epitopes and how they could influence CD. The gliadin-induced activation of the 'innate' immune system might also have a significant role in the induction and persistence of many CD complications and most definitively for the most aggressive one, namely mucosal T cell lymphomas. We further suggest a novel way to harness the unwanted immune response to toxic prolamine, and thus indicate new potential therapeutic strategies to treat or at least control CD. PMID- 15829282 TI - Mannose-binding lectin: do we need it? AB - Mannose-binding lectin (MBL) deficiency has now been recognised for 15 years. Numerous studies have shown that MBL deficiency is associated with an increased susceptibility to a range of infections. However, the importance of MBL, in defence against infection, is still debated. This article discusses recent developments in MBL research and explores how MBL may be operating in the setting of modern medical practice. PMID- 15829283 TI - Innate immunity: past, present and future. PMID- 15829284 TI - CRP after 2004. AB - C-reactive protein (CRP) that has been conserved throughout evolution is a host defense molecule. Its attraction towards phosphocholine-ligands, such as modified low-density lipoprotein, and apoptotic cells leads to the "masking" of these substances that have the capabilities to otherwise engage in deleterious activities. Complement activation by CRP complexes and the modulation by CRP of complement activation by its ligands add up to its beneficial effects. In the presence of CRP, production of membrane-damaging last product of the complement pathway is arrested. CRP is currently serving as an indicator of cardiovascular diseases, but to pinpoint the role of CRP in atherosclerosis, a drug that can lower cholesterol levels, but not the CRP levels, is needed for experimentation. PMID- 15829285 TI - Anticipating innate immunity without a Toll. AB - Earthworm innate immunity depends upon small and large leukocytes (coelomocytes) that synthesize and secrete humoral antimicrobial molecules (e.g. lysenin, fetidin, eiseniapore, coelomic cytolytic factor [CCF]; Lumbricin I). Small coelomocytes (cytotoxic) are positive (CD11a, CD45RA, CD45RO, CDw49b, CD54, beta(2)-m and Thy-1 [CD90]; CD24; TNF-alpha) but negative using other mammalian markers. Large coelomocytes (phagocytic) are uniformly negative. Specific earthworm anti-EFCC 1, 2, 3, 4 mAbs are negative for Drosophila melanogaster hemocytes and mammalian cells but positive those of earthworms. Coelomocytes contain several lysosomal enzymes involved in phagocytosis and a pattern recognition molecule (CCF) that may trigger the prophenoloxidase cascade a crucial innate immune response. Earthworms and other invertebrates possess natural, non-specific, non-clonal, and non-anticipatory immune response governed by germ line genes. Toll and Toll-like receptor signaling is essential for phagocytosis and antimicrobial peptide synthesis and secretion in insects and vertebrates but has not yet been shown to be essential in earthworm innate responses. PMID- 15829286 TI - Despite extensive similarity in germline DH and JH sequence, the adult Rhesus macaque CDR-H3 repertoire differs from human. AB - Rhesus macaques (Macaca mulatta) and chimpanzees (Pan troglodytes) are frequently used models for the study and treatment of human infectious diseases, including AIDS. Confidence in the equivalence to human of the humoral immune responses of these higher primates has grown as studies of immunoglobulin germline sequences have documented average identities of 90% or greater to human counterparts. The most variable component of the immunoglobulin heavy chain, complementarity determining region 3 (CDR-H3) is the product of somatic (junctional) as well as germline (combinatorial) mechanisms of diversity. Located at the center of the antigen binding site, CDR-H3 often exerts a dominant role in antibody specificity and affinity. To test whether similarity in germline DH and JH sequence would yield similarity in CDR-H3 composition in the expressed repertoire, we compared IgM CDR-H3 transcripts from Rhesus and chimpanzee blood to human. In fetal Rhesus, the range and mean of CDR-H3 lengths was similar to that observed in fetal human. However, the Rhesus repertoire of adult muCDR-H3 transcripts did not contain the longer hypervariable intervals that humans begin to express late in the second trimester of fetal life. Conversely, the adult chimpanzee repertoire included more long CDR-H3 structures than human. The differences between these adult repertoires reflected fine changes in N addition and terminal nucleotide loss. We conclude that the same mechanisms that refine and shape CDR-H3 diversity during ontogeny can also be used to fine tune and individualize species-specific antibody repertoires despite germline immunoglobulin sequence similarity. PMID- 15829287 TI - Toll-like receptor mRNA expression patterns in human dendritic cells and monocytes. AB - The innate immune system recognises a wide spectrum of pathogens without a need for prior exposure. The main cells responsible are monocytes, macrophages, dendritic cells (DC) and neutrophils phagocytose microbial pathogens triggering a cytokine network resulting in the development of inflammatory and specific immune responses. Findings in the Toll-like receptor (TLR) family, initially discovered in Drosophila, further elucidated these processes. Toll-like receptors induce activation of an innate immune response and at present ten TLRs have been identified, named TLRs 1-10. In addition to the ignition of the innate immune response, evidence implicates the TLR family in a spectrum of systemic disorders following bacterial infections including sepsis and multiple organ failure, and can be detrimental, leading to tissue injury. In this project, our main goal was to investigate the effects of a TLR4 ligand, lipolysaccharide (LPS) in human DC and monocytes. Our hypothesis is that different professional APCs, express different mRNA TLR transcripts. Our findings indicate that TLR expression patterns change in relation to the pathogen involved and in the case of DC, and the maturation stage the latter are upon challenging. Our results and interpretation showed significant alteration of transcript expression patterns upon LPS challenge in all cell subsets, with DC subsets expressing different TLR mRNA patterns as they go through different maturation stages. PMID- 15829288 TI - Analysis of mannose-binding lectin 2 (MBL2) genotype and the serum protein levels in the Korean population. AB - Mannose-binding lectin (MBL) is a C-type lectin produced by the liver and involved in the innate immune response. We have analyzed six SNPs of MBL2 gene- three at promoter (-550, -435, and -221), one at 5'-untranslational region (UTR) (+4), and two at coding (Gly54Asp and Leu126Leu) regions--in the Korean population (N=129), and have correlated genotypes with the serum concentration and functional characteristics. Of those, the Asp54 allele (P<10(-15)), L allele at -550 (P<10(-7)), and P allele at +4 (P=0.012) were correlated with low MBL levels. The effect of the X allele at -221 on MBL levels in the Korean population appeared to be less profound than that of other populations. The highest MBL producing promoter haplotype in the Korean population was HYP, followed by LYQ and LYP, and then LXP. From functional analysis of MBL, low MBL levels were correlated with low mannan-binding, low C4 complement activation, and lack of high ordered oligomers. Our results support that the promoter and coding polymorphisms of MBL are correlated with its functional activity as well as circulating levels, and the association patterns are quite similar to those of other populations. PMID- 15829289 TI - Single chain antibody fragments for the selective targeting of antigens to dendritic cells. AB - In order to target antigens (Ags) selectively to dendritic cells (DC), we derived single chain antibody fragments (scFvs) from NLDC-145 and N418, two monoclonal antibodies binding the mouse dendritic cell-restricted surface molecules DEC-205 and CD11c. Recombinant hexahistidine-tagged forms of the scFvs (scNLDC and scN418) were efficiently produced in a baculovirus expression system. Both scFvs bound DEC-205(+) Langerhans cells and CD11c(+) fetal skin-derived dendritic cells (FSDCs) comparably to their parental antibodies. Immunization of C57BL/6 mice with a DNA vaccine encoding a model protein antigen fused to scNLDC stimulated specific immune responses in both the humoral and cellular compartments, in contrast to DNA vaccines expressing scN418-targeted or untargeted antigen. Our results show that antigen targeting to DCs via a DEC-205 binding scFv leads to enhanced immunogenicity. Further, this work suggests that scFvs fused to protein antigens and delivered as DNA vaccines may provide a generic means for delivering vaccinal molecules to selected cell populations. PMID- 15829290 TI - L-gamma-Glutamyl-L-cysteinyl-glycine (glutathione; GSH) and GSH-related enzymes in the regulation of pro- and anti-inflammatory cytokines: a signaling transcriptional scenario for redox(y) immunologic sensor(s)? AB - Of the antioxidant/prooxidant mechanisms mediating the regulation of inflammatory mediators, particularly cytokines, oxidative stress-related pathways remain a cornerstone. It is conspicuous that there is a strong association between free radical accumulation (ROS/RNS; oxidative stress) and the evolution of inflammation and inflammatory-related responses. The scenario that upholds a consensus on the aforementioned is still evolving to unravel, from an immunologic perspective, the molecular mechanisms associated with ROS/RNS-dependent inflammation. Cytokines are keynote players when it comes to defining an intimate relationship among reduction-oxidation (redox) signals, oxidative stress and inflammation. How close we are to identifying the molecular basis of this intricate association should be weighed against the involvement of specific signaling molecules and, potentially, transcription factors. L-gamma-Glutamyl-L cysteinyl-glycine, or glutathione (GSH), an antioxidant thiol, has shaped, and still is refining, the face of oxidative signaling in terms of regulating the milieu of inflammatory mediators, ostensibly via the modulation (expression/repression) of oxygen- and redox-responsive transcription factors, hence termed redox(y)-sensitive cofactors. When it comes to the arena of oxygen sensing, oxidative stress and inflammation, nuclear factor-kappaB (NF-kappaB) and hypoxia-inducible factor-1alpha (HIF-1alpha) are key players that determine antioxidant/prooxidant responses with oxidative challenge. It is the theme therein to underlie current understanding of the molecular association hanging between oxidative stress and the evolution of inflammation, walked through an elaborate discussion on the role of transcription factors and cofactors. Would that classify glutathione and other redox signaling cofactors as potential anti inflammatory molecules emphatically remains of particular interest, especially in the light of identifying upstream and downstream molecular pathways for conceiving therapeutic, alleviating strategy for oxidant-mediated, inflammatory related disease conditions. PMID- 15829291 TI - Structure-based design and characterization of a Novel IL-6 antagonist peptide. AB - The development of rational methods to design antagonist peptides based on the 3 D structure of protein active region has, to now, been only marginally successful. This has been largely due to the difficulty of constraining the recognition elements of a mimetic structure to the relative conformational and spatial orientations present in the parent molecule. According to the 3-D complex structure of human interleukin-6 (hIL-6) and its receptor (hIL-6R), a novel antagonist peptide (named PT), which possessed potential bioactivity of hIL-6, was designed by the means of distance geometry, molecular modeling and molecular dynamics trajectory analysis. The bioactivity of the designed peptide (i.e. PT) was evaluated using XG-7 cells, a hIL-6-dependent B-cell line. PT possessed potential bioactivity to antagonize the function of hIL-6 and could efficiently induce the growth arrest and apoptosis of XG-7 cells in a dose-dependent manner. PMID- 15829292 TI - Tumor-induced suppression of interferon-gamma production and enhancement of interleukin-10 production by natural killer (NK) cells: paralleled to CD4+ T cells. AB - The predominance of type two cytokines in syngeneic B16 tumor-bearing mice was confirmed by analysing supernatant contents and mRNA copies of IFN-gamma, IL-4, IL-5, IL-10 and IL-13 from splenocytes. The cytokine-producing lymphocytes were then examined by double-staining flowcytometry. Both CD4+IFN-gamma+ T cells and DX5+IFN-gamma+ NK cells from spleen significantly declined, interestingly, the declining degrees of DX5+IFN-gamma+ NK cells were much greater than those of CD4+IFN-gamma+ T cells by the percentage in whole NK or T cells or the absolute amounts per spleen at early tumor stage (day 10) or tumor-advanced stage (day 20). In contrast to DX5+IFN-gamma+ NK cells, DX5+IL-10+ NK cells increased during tumor progression, the increasing degrees of DX5+IL-10+ NK cells were also much greater than those of CD4+IL-10+ T cells by the percentage or the absolute amounts. Though the percentage of DX5+IL-4+ NK cells only increased in early tumor stage (day 10), the increasing degree was also greater than that of CD4+IL 4+ T cells. In 20xfield view under laser confocal microscope, the mean numbers of DX5+IFN-gamma+ NK cells and CD4+IFN-gamma+ T cells dramatically declined after tumor inoculation. These results suggest that cytokines produced by NK cells, at least partly, account for the balance of type one and two cytokines as done by T cells, and in some conditions, that the NK1 or NK2 cells were possibly more sensitive to tumor progression. PMID- 15829293 TI - Apoptotic and necrotic cells induced by different agents vary in their expression of MHC and costimulatory genes. AB - We have recently reported, in a murine tumor model, that apoptotic cells induced by different agents may vary in their ability to elicit host immunity. The basis for this observation is unclear but may involve varying efficiencies of cross presentation and/or direct activation of immunity by different apoptotic preparations. As a first step in addressing this issue, we compared expression patterns of selected immune genes (MHC class I, class II, CD40, B7-1, B7-2) on viable and apoptotic populations induced by four different agents. The histone deacetylase inhibitor trichostatin A (TSA) induced MHC class II expression on viable and apoptotic cell populations, while LPAM, H2O2 and gamma-irradiation did not activate class II. Each agent employed elicited a different expression pattern of costimulatory molecules (CD40, B7-1, B7-2) on both apoptotic and 7 AAD+ 'necrotic' populations. In striking contrast to the TSA induction of MHC class II, class I cell surface protein was diminished on the apoptotic populations. These effects were not a result of changes in the cell cycle produced by the various treatments. The data demonstrate that distinctive gene expression patterns on viable and apoptotic cells are elicited by different apoptosis inducing agents. We discuss how expression patterns on dead or dying tumor cells could potentially affect the tumor's ability to elicit immunity. PMID- 15829294 TI - Hybridization signatures during thymus ontogeny reveals modulation of genes coding for T-cell signaling proteins. AB - Non-manipulated inbred mouse strains constitutes an interesting model-system for in vivo studies on thymus ontogeny due to the possibility to observe the molecular events of the thymocyte maturation. In previous studies, using RT-PCR method, we have found that several immune system genes such as interleukins and MHC are differentially expressed during ontogeny of the thymus whose genes act as modulators of T-cell differentiation. To determine which other genes are modulated on a large-scale basis, we measured the levels of mRNA expression in mouse fetal thymus (14-17 days of gestation) by hybridization with cDNA microarrays containing 1,576 cDNA sequences derived from the IMAGE MTB library. T cell maturation was monitored by detection of the T-cell receptor beta TRBV8.1 BD2.1 rearranged DNA segment. Each developmental phase of thymus, displayed a characteristic expression profile, as evaluated by the Cluster and Tree-View softwares. Genes differentially and significantly expressed were selected on the basis of significance analysis of the microarray data (SAM program). With the reclustering of only significantly expressed genes, it was possible to characterize the phases of thymus ontogeny, based on the differential profile of expression. Our method provided the detection of genes implicated in the cell signaling, such as the hematopoietic cell signal transducer gene, genes implicated in T-cell calcium influx (tyrosine phosphatase) and calcium signaling proteins (vesicle transport binding protein 3, proline rich Gla, casein kinase alpha 1 and Down syndrome homolog protein 1) and a gene important for the protein transport, including T-cell receptors chains, towards the cell membrane (Golgi SNAP receptor complex member 2). The results demonstrate that the cDNA microarray used to explore the gene expression was useful for understanding the modulation of several cell-signaling genes, including the calcium cascade pathway, which is important for individual stages of T-cell maturation and control of anergy during thymus ontogeny. PMID- 15829295 TI - Differential induction of apoptosis by LPS and taxol in monocytic cells. AB - Numerous microbial as well as other stimulants including lipopolysaccharide and taxol can activate TLR4, and elicit diverse downstream signaling events including cytokine gene expression and cell growth regulation. With a mechanism not completely understood, different TLR4 stimulants induce distinct cellular responses. Our present studies showed that taxol, not LPS, induced cell apoptosis in human monocytic THP-1 cells, as indicated by PARP cleavage, as well as bcl-2 phosphorylation. Pretreatment of cells with LPS abolished subsequent taxol effect, suggesting that certain signaling components involved in taxol-mediated apoptosis were disrupted by LPS pretreatment. Since the decrease in IRAK-1 level closely accompanies prolonged LPS treatment in monocytic cells, we investigated the IRAK-1 status upon various taxol and LPS challenges. We observed that only LPS, not taxol, caused dramatic decrease in IRAK-1 protein levels. Using splenic macrophages harvested from IRAK-1 knockout and control mice, we further demonstrated that the presence of IRAK-1 is required for taxol-induced PARP cleavage. PMID- 15829296 TI - Functional characterization of full-length TLR3, IRAK-4, and TRAF6 in zebrafish (Danio rerio). AB - Recently, the zebrafish, Danio rerio, has been recognized as a useful model for infectious disease and immunity. The Toll-like receptor (TLR) family is an evolutionarily conserved component of the innate immune system that responds to specific pathogen-associated molecular patterns (PAMPs) during an infection. This study reports the identification and characterization of a full-length orthologue of mammalian TLR3, and the key TLR pathway signaling molecules IRAK-4 and TRAF6 in the zebrafish. Sequence analysis of zebrafish TLR3 (zfTLR3), IRAK-4 (zfIRAK 4), and TRAF6 (zfTRAF6) revealed conserved domains shared with insect and mammalian genes. Quantitative real-time PCR showed that all three genes are expressed in a variety of adult tissues and during embryonic development. In in situ hybridization, we showed that zfTLR3, zfIRAK-4, and zfTRAF6 are present in distinct regions of the developing brain at 22hpf and that zfTRAF6 was observed in the developing medial neural tube. Overexpression of zfIRAK-4, zfTRAF6, or a mutant zfTLR3 construct was able to stimulate NF-kappaB activation in ZFL cells as measured by a cotransfected NF-kappaB-luciferase reporter plasmid. Messenger RNA expression profiles of each gene in zebrafish embryos and adults were examined by quantitative real-time PCR following infection with snakehead rhabdovirus (SHRV) or Edwardsiella tarda. Following exposure to SHRV, only zfTLR3 and zfTRAF6 mRNA transcripts were upregulated. Interestingly, exposure of fish to E. tarda resulted in an unexpected increase in mRNA expression of zfTLR3, as well as the anticipated upregulation of zfIRAK-4 and zfTRAF6 mRNA transcripts. These results demonstrate that zebrafish possess conserved TLR-signaling pathways, further emphasizing the utility of the zebrafish as a model for vertebrate immunology. PMID- 15829297 TI - Correlation between beta-defensin expression and induction profiles in gingival keratinocytes. AB - Human beta-defensins are antimicrobial peptides produced by epithelial cells. To date, 28 beta-defensins have been described and the expression of a select few has been classified as constitutive or inducible. Most studies have evaluated expression and regulation using a limited number of primary cell cultures or immortalized cell lines. The goal of this study was to quantitatively assess the in vitro expression and inducibility profiles of human beta-defensins, HBD-1, HBD 2, and HBD-3 across a number of primary gingival keratinocyte cultures. Cultured cells from 14 human subjects were stimulated with interleukin-1 beta (IL-1beta), IL-2, IL-6, IL-8, IL-12, tumor necrosis factor alpha (TNF-alpha), gamma interferon (IFN-gamma) or Escherichia coli lipopolysaccharide (LPS) and analyzed by reverse transcription (RT)-PCR. A subset of cultures were quantitatively assessed by real-time PCR. HBD-1 presented the highest and most heterogeneous expression at the basal level (non-stimulated) as compared to expression of HBD-2 and HBD-3, which was significantly lower and homogeneous. IFN-gamma was a primary inducer for HBD-1 and HBD-3, while IL-1beta and TNF-alpha were primary inducers for HBD-2. Sporadic induction was seen for IL-2, IL-6 and LPS. Synergistic expression was seen when various cytokines were combined. Interestingly, the induction potential of each beta-defensin was directly correlated to its basal expression. An inhibitor of JAK2 kinase (Janus kinase), down-regulated IFN-gamma induced HBD-1 and HBD-3 expression, suggesting a role for the JAK/signal transducer and activator of transcription (STAT) signaling pathway in their expression. HBD-2 protein expression of supernatants and cell lysates paralleled mRNA expression. The results suggest that beta-defensin expression and induction in gingival keratinocytes is similar to that seen in other tissue. However, the novel finding of considerable variation among induction levels and the correlation of the induction with basal expression suggests that these innate response elements may play a key role in susceptibility or resistance to disease in the oral cavity. PMID- 15829298 TI - Structural analysis of substitution patterns in alleles of human immunoglobulin VH genes. AB - The diversity in repertoires of antibodies (Abs) needed in response to the antigen challenge is produced by evolutionary and somatic processes. The mechanisms operating at a somatic level have been studied in great detail. In contrast, neither the mechanisms nor the strategies of diversification at an evolutionary level have yet been understood in similar detail. Particularly, the substitution patterns in alleles of immunoglobulin genes (Igs) have not been systematically studied. Furthermore, there is a scarcity of studies which link the analysis at a genetic level of the diversification of repertoires with the structural consequences at the protein level of the changes in DNA information. For the purpose of systematically characterizing the strategies of evolutionary diversification through sequence variation at alleles, in this work, we built a database for all the alleles of the IGHV locus in humans reported until now. Based on these data, we performed diverse analyses of substitution patterns and linked these results with studies at the protein level. We found that the sequence diversification in different alleles does not operate with equal intensity for all V genes. Our studies, both of the number of substitutions and of the type of amino acid change per sub-segment of the V-REGION evidenced differences in the selective pressure to which these regions are exposed. The implications of these results for understanding the evolutionary diversification strategies, as well as for the somatic generation of antibody repertoires are discussed. PMID- 15829299 TI - HIV-1 neutralization by chimeric CD4-CG10 polypeptides fused to human IgG1. AB - The envelope glycoprotein of HIV-1 is the principal target for entry inhibitors. The use of soluble CD4 has been found to be impractical as most clinical isolates are resistant to neutralization at feasible concentrations. CG10 is one of a small group of monoclonal antibodies specific to CD4-induced epitopes, which are structurally associated with the chemokine receptor-binding site and are capable of blocking the interaction of gp120 with its obligatory co-receptor. We have reasoned that fusing the single chain Fv of CG10 with CD4 can lead to increased HIV-1 neutralization activity and that this effect could be further enhanced by engrafting this chimeric construct onto an IgG Fc. Here we report the cloning of the genes encoding the variable regions of CG10 heavy and light chains and demonstrate that when attached to human IgG1 Fc, the single chain Fv of CG10 retains the binding properties of the original mouse antibody. Fusing CG10 single chain Fv with the gp120-binding portion of CD4 on a human IgG1 Fc backbone results in stronger binding of gp120 of different tropisms and in enhanced neutralization of laboratory-adapted strains and most, but not all, clade B and clade C isolates tested. Our findings underscore the potential use of CD4-based fusion proteins in the design of HIV immuno-therapeutics. PMID- 15829300 TI - A point mutation in the Ch3 domain of human IgG3 inhibits antibody secretion without affecting antigen specificity. AB - Immunoglobulins (Ig) require correct folding and assembly of both heavy (H) and light (L) chains to form a functional H2L2 dimer that is secreted from plasma cells. This process is dependent upon the endoplasmic reticulum (ER) chaperone BiP, which targets improperly, folded or assembled Ig molecules for degradation. While investigating the mechanism of low IgG3 secretion, we identified a missense mutation L368P in the Ch3 region of the human gamma3 H-chain that was associated with impaired secretion of intact and functional Ig. The non-secreted H-chains displayed slower electrophoretic migration than secreted H-chains, consistent with them being glycosylated in the ER but not fully processed in the golgi apparatus and secretory pathway. Reversion of the mutated codon to wild type restored secretion of the IgG3, which displayed the same fine specificity for antigen as non-secreted IgG3. However, the non-secreted IgG3 was not opsonic in an in vitro phagocytosis assay. The results indicate that correct IgG3 Ch3 domain folding is essential for secretion and effective function but does not affect specificity for antigen. PMID- 15829301 TI - Matching of trastuzumab (Herceptin) epitope mimics onto the surface of Her-2/neu- a new method of epitope definition. AB - As seen with the proto-oncogene Her-2/neu, antibodies targeting different parts of a receptor can have opposing effects. Depending on epitope specificity, in this case, tumor growth can be inhibited--but also enhanced. Therefore, the definition of molecular binding sites is of increasing importance in modern medicine. We here introduce a novel approach for binding site localization, utilizing information obtained by the phage display technique. This is a high throughput screening method for identification of peptide mimics, so called mimotopes, of any binding structure of interest. All target molecules whose structure is available in the RCSB Protein Data Bank can be scanned for mimotope matches on their surface. In this study, we present the matching results of five mimotopes defined for the epitope recognized by trastuzumab (Herceptin), a humanized monoclonal antibody inhibiting tumor growth, on Her-2/neu. The localization thus obtained corresponds to the known trastuzumab epitope. We therefore suggest the algorithm as a novel way of binding site definition, circumventing co-crystallization experiments. PMID- 15829302 TI - Might there be a link between mannose-binding lectin polymorphism and dental caries? AB - The aim of this study is to analyze two polymorphisms in exon 1 of Mannose binding lectin (MBL) gene in children with carious teeth and children caries-free in order to determine the frequencies of these polymorphisms and to investigate possible association between MBL polymorphisms and dental caries. Fourty-two children with carious teeth and 40 children caries-free participated in the study. Two-hundred microliters of peripheral blood samples were taken in EDTA tubes and genomic DNA was isolated. PCR-RFLP method was used with BanI and MboII digestion enzymes. The overall distribution of genotypes did not significantly differ between two groups and there was also no significant difference in the allele frequency of codon 54 wild type (allele A) (p=0.884, p=0.585). It has been concluded that further investigations may be required to show possible association between MBL and dental caries in which high number healthy children are participated. PMID- 15829303 TI - Mixed-haplotype MHC class II molecules select functional CD4+ T cells. AB - MHC class II molecules are formed from polymorphic alpha and beta chains. While pairing of chains is most efficient within class II isotypes and haplotypes, limited pairing and surface expression of mixed-haplotype and -isotype class II molecules is common. The function of such molecules in antigen presentation has been established by the unique restriction of responses in F1 mice. However, it has not been established whether mixed class II molecules are able to mediate selection of functional T cells and how the reduced avidity of the TCR/MHC interaction influences the repertoire. In this report we have addressed these issues through the production of mice expressing solely mixed-haplotype class II molecules. The mixed class II molecules promote selection of a small CD4+ T cell repertoire with modified TCR use. The selected CD4+ T cells are functional in vivo and in vitro. PMID- 15829304 TI - Oxiagin from the Naja oxiana cobra venom is the first reprolysin inhibiting the classical pathway of complement. AB - A basic glycoprotein oxiagin with molecular mass of 49.8 kDa was isolated from the venom of Central Asian cobra Naja oxiana. Partial amino acid sequence determination has shown that oxiagin belongs to reprolysins, a subfamily of animal metalloproteinases possessing a characteristic multidomain structure. Oxiagin was found to inhibit the classical pathway of the complement system. A study of the oxiagin influence on the different stages of the classical pathway showed that it inhibited the formation of C3-convertase. To achieve it, oxiagin binds to IgG on the surface of sheep erythrocytes sensitized with rabbit antibodies, thus, preventing the interaction of component C2 (without its inactivation) with immobilized C4b. IC50 for the inhibiton of classical pathway of complement system by oxiagin is 80 nM, while it does not affect the alternative pathway at concentrations up to 1.2 microM. Oxiagin possessed hemagglutinating activity towards sheep and rabbit erythrocytes, and this activity as well as the complement inhibition by oxiagin were suppressed by D galactose. Oxiagin is the first representative of snake venom reprolysins that inhibits the complement system, utilizing non-proteolytic inhibiting strategy. PMID- 15829305 TI - The effects of anti-histone H1 antibody on immune cells responsible for rejection reaction. AB - We previously demonstrated the immunosuppressive activity of anti-histone H1 autoreactive antibodies (Ab) transiently induced in serum of a rat tolerogenic orthotopic liver transplantation (OLT) model. In the present study, we investigated the effects of anti-histone H1 Ab on dendritic cells (DCs), T-cells, lymphokine-activated killer (LAK) cells, and human natural killer (NK) cells. The effects of anti-histone H1 Ab on Concanavalin A (ConA) blast, on rat DC cytokine profiles and phenotypes, and on T-cells, LAK cells, and human NK cells were examined by flow cytometry and RT-PCR. The cytotoxicity of LAK and NK cells pretreated with anti-histone H1 Ab was assayed. The addition of anti-histone H1 Ab to ConA blast inhibited the proliferation of 5-(6)-carboxy-fluorescein succinimidyl ester (CFSE)-labeled lymphocytes without toxicity but increased the population of CD4+CD25+ T-cells. DCs treated with anti-histone H1 Ab expressed lower levels of CD80/CD86, IL-1beta, and IL-6. The addition of anti-histone H1 Ab to LAK culture decreased the percentages of NKR-P1 populations and down-regulated levels of inducible nitric oxide synthase (iNOS), IL-2, and INF-gamma in RT-PCR. The cytotoxicity of LAK and NK cells was lower when pretreated with anti-histone H1 Ab than when pretreated with control IgG. We found that the blockade of histone H1 modulated DCs toward tolerogenic status, decreased the cytotoxicity of LAK and NK cells, and induced CD4+CD25+ T-cells. These results suggest that the use of anti-histone H1 Abs might be a useful strategy for the development of a form of immunosuppression. PMID- 15829306 TI - Pyrrolidine dithiocarbamate-induced macrophage inflammatory protein-2 gene expression is NF-kappaB-independent but c-Jun-dependent in macrophage cell line RAW 264.7. AB - Pyrrolidine dithiocarbamate (PDTC) is a stable compound that acts as antioxidant or prooxidant, and is widely used to inhibit the activation of NF-kappaB. PDTC was also reported to activate NF-kappaB depending on its dose and metal ions in PC12 cells. In this work, we demonstrated a working mechanism of PDTC and its effects on the proinflammatory cytokine gene expression in a mouse macrophage cell line, RAW 264.7. PDTC alone induced NF-kappaB-independent MIP-2 promoter activation that can be assessed by transient transfection and confocal image analysis. The involvement of AP-1 transcription factor was noticed by promoter deletion/site-specific mutation analysis and electrophoretic mobility shift assay (EMSA). Among three different mitogen-activated protein kinase (MAPK) pathways tested, only the stress-activated protein kinase (SAPK)/Jun N-terminal kinase (JNK) pathway was significantly activated in RAW 264.7 cells after the stimulation with PDTC. Using pathway-specific inhibitors, we found that the SAPK/JNK pathway is clearly associated with PDTC-induced MIP-2 gene expression. Our experimental results indicate that PDTC-induced proinflammatory cytokine expressions are mediated by SAPK/JNK pathway, which activates AP-1. PMID- 15829307 TI - Phosphatidylinositol-3-kinase regulates PKCtheta activity in cytotoxic T cells. AB - Protein kinase C (PKC) theta plays a crucial role in T cell activation. We, therefore, examined the regulation of PKCtheta activity in cytotoxic T lymphocytes (CTL). We demonstrated that PMA did not stimulate PKCtheta activation and phospholipase C inhibition did not block anti-CD3-stimulated PKCtheta activation in a CTL clone. This suggests that diacylglycerol is neither sufficient nor required for PKCtheta activation. Furthermore, PKCtheta was only activated in a CTL clone stimulated with plate-bound anti-CD3 but not soluble anti-CD3. However, PMA or cross-linked anti-CD3 stimulated phosphorylation of PKCtheta as measured by a migratory shift, suggesting that phosphorylation was not sufficient for activity. Phosphatidylinositol 3-kinase activity was required for anti-CD3, but not PMA, stimulated phosphorylation and for immobilized anti CD3-triggered PKCtheta activity. A substantial fraction of PKCtheta was constitutively membrane associated and PMA or CD3 stimulation did not significantly increase membrane association. Our data indicate that phosphorylation of PKCtheta is not a suitable surrogate measurement for PKCtheta activity and that additional, yet to be defined steps, are required for the regulation of PKCtheta enzymatic activity in CTL. PMID- 15829308 TI - Transcriptome profiling of adult zebrafish at the late stage of chronic tuberculosis due to Mycobacterium marinum infection. AB - The Mycobacterium marinum-zebrafish infection model was used in this study for analysis of a host transcriptome response to mycobacterium infection at the organismal level. RNA isolated from adult zebrafish that showed typical signs of fish tuberculosis due to a chronic progressive infection with M. marinum was compared with RNA from healthy fish in microarray analyses. Spotted oligonucleotide sets (designed by Sigma-Compugen and MWG) and Affymetrix GeneChips were used, in total comprising 45,465 zebrafish transcript annotations. Based on a detailed comparative analysis and quantitative reverse transcriptase PCR analysis, we present a validated reference set of 159 genes whose regulation is strongly affected by mycobacterial infection in the three types of microarrays analyzed. Furthermore, we analyzed the separate datasets of the microarrays with special emphasis on the expression profiles of immune-related genes. Upregulated genes include many known components of the inflammatory response and several genes that have previously been implicated in the response to mycobacterial infections in cell cultures of other organisms. Different marker genes of the myeloid lineage that have been characterized in zebrafish also showed increased expression. Furthermore, the zebrafish homologs of many signal transduction genes with relationship to the immune response were induced by M. marinum infection. Future functional analysis of these genes may contribute to understanding the mechanisms of mycobacterial pathogenesis. Since a large group of genes linked to immune responses did not show altered expression in the infected animals, these results suggest specific responses in mycobacterium-induced disease. PMID- 15829309 TI - An HLA-E single chain trimer inhibits human NK cell reactivity towards porcine cells. AB - HLA-E, when expressed by pig cells, could alleviate human natural killer (NK) cell-mediated rejection of porcine xenografts by providing a potent inhibitory ligand for human NK cells expressing CD94/NKG2A. Yet cell-surface expression of HLA-E on porcine epithelial (LLC-PK1) cells was not observed after transfection with an expression vector harboring HLA-E alone or in combination with an expression vector containing human beta2m. A single chain trimer (SCT) of HLA-E consisting of, in the following order (from N- to C-terminus), the leader peptide of human beta2m, VMAPRTLIL (an HLA-E-binding peptide), a 15 amino acid linker, mature human beta2m, a 20 amino acid linker, and mature HLA-E heavy chain was engineered. Cell-surface expression and correct folding of HLA-E SCT was shown by FACS analyses of stably transfected LLC-PK1 cells. Untransfected LLC-PK1 cells were readily lysed by the NK cell lines NKL and NK-92, while LLC-PK1 cells expressing HLA-E SCT were almost completely protected. In addition, the HLA-E SCT recapitulates the peptide dependent properties of normal HLA-E trimeric complexes in that an HLA-E SCT with an hsp60 derived peptide, though expressed at the cell surface, did not inhibit NK cell-mediated lysis. The HLA-E SCT, which conferred protection against NK cell-mediated killing, also inhibited NK cell IFN-gamma secretion elicited by co-culture of NKL cells with LLC-PK1 cells. Thus, HLA-E SCT, in which all three components of a normal HLA-E protein complex are in one polypeptide chain, is immunologically functional as it is able to modulate NK cell cytotoxicity and cytokine secretion. PMID- 15829310 TI - Activation of rainbow trout (Oncorhynchus mykiss) mononuclear phagocytes by different pathogen associated molecular pattern (PAMP) bearing agents. AB - Rainbow trout (Oncorhynchus mykiss) cells of a monocyte-macrophage lineage (rtMOCs) were used to characterize the ability of the trout innate immune system to recognize and respond to different pathogen associated molecular pattern (PAMP) bearing substances. Compared to what has been reported for mammalian macrophages, rtMOCs responded with lower sensitivity to lipopolysaccharide (LPS) from Escherichia coli (EC-LPS) and Pseudomonas aeruginosa (PA-LPS). The sensitivity of rtMOCs to LPS was not influenced by the presence of serum which suggests that the resistance to endotoxic shock in fish may be due to the lack of serum-borne factors that confer sensitivity to LPS in mammals. The time course of the response to PAMPs could be separated into two patterns. EC-LPS induced stable cytokine expression whereas PA-LPS, zymosan and muramyl dipeptide induced transient TNF2 expression. By analogy to the type of stimulation observed in mammals it can be hypothesized that different signaling pathways, possibly initiated by different receptors, may be involved in the recognition of these PAMPs by rtMOCs. PMID- 15829311 TI - Characterisation of salmon and trout CD8alpha and CD8beta. AB - The genes and corresponding cDNAs of both alpha and beta chains of the Atlantic salmon (Salmo salar) CD8 molecule have been sequenced and characterized. In addition, the cDNAs for alpha and beta chains of brown trout (Salmo trutta) and for the beta chain in rainbow trout (Oncorhynchus mykiss) have been sequenced. The cDNAs code for signal sequences which are preceded by short 5' UTRs. These are followed by typical immunoglobulin superfamily variable sequences all of which contain two conserved cysteines for the intra-chain disulphide bond. The hinge regions display conserved cysteines for dimerisation and several O glycosylation motifs for each predicted protein. The domain sharing the highest sequence identity with mammals is the single pass transmembrane domain for all sequences. In salmon, each domain is predominantly coded for by a single exon except the cytoplasmic/3' UTR domains, which are coded for by 3 and 2 exons for the alpha and beta genes, respectively. In the alpha gene, the second cytoplasmic exon may be spliced out to form an alternative shorter transcript which if expressed would exhibit a truncated cytoplasmic tail. A splice variant found for the salmon beta gene introduces a stop codon after only 40 amino acids. Overall amino acid identities between salmonid sequences were higher than 90%, whereas they shared only 15-20% identity with species such as, chicken and human. Analysis of the expression patterns of the two salmon genes using quantitative RT PCR shows a very high expression in the thymus. This is mirrored by the expression of the TCRalpha gene, which is known to be co-expressed with CD8 on mammalian T cells. This is the first report of a sequence for CD8beta in a teleost and together with the CD8alpha sequence, it encodes the ortholog of the CD8 co-receptor molecule on mammalian T cells. PMID- 15829312 TI - Proviral integration of an Abelson-murine leukemia virus deregulates BKLF expression in the hypermutating pre-B cell line 18-81. AB - The transcription factor BKLF (basic Kruppel-like factor, KLF3) is a member of the Kruppel-like factors (KLF) family. KLF members harbor a characteristic C terminal zinc-finger DNA-binding domain and bind preferentially to CACCC-motifs. BKLF is highly expressed in haematopoietic and erythoid cells and works either as repressor or activator of transcription in various genes. BKLF-deficient mice display myeloproliferative disorders and abnormalities in haematopoiesis. Other members of the KLF-family such as GKLF and BCL11A have been implicated in tumorigenesis, however, for BKLF such association has not yet been demonstrated. We report here that a single Abelson-murine leukemia virus (A-MuLV) provirus is present in the genome of the hypermutating murine pre-B cell line 18-81. The provirus has integrated into the locus of the transcription factor BKLF. In contrast to other A-MuLV transformed pre-B cell lines, BKLF is highly transcribed in cell line 18-81. BKLF transcripts originate from the retroviral long terminal repeats (LTRs) and BKLF protein can be detected by gel shift retardation assay. We hypothesize on a potential role of BKLF deregulation in tumorigenesis and/or in the induction of somatic hypermutation in cell line 18-81. PMID- 15829313 TI - Analysis of recombination signal sequences in zebrafish. AB - Recombination signal sequences (RSS) from immunoglobulin and TCRalpha genes of zebrafish were analyzed in comparison with RSS from human and species-specific features were revealed. In contrast to human RSS, in zebrafish RSS from both V(H) and TCRalpha genes the last nonamer position is not conserved. On the contrary, the fourth nonamer position, which is not conserved in human or mouse is conserved in zebrafish. The 12 bp spacers from human and zebrafish RSS contain 9 bp motif resembling nonamer sequence. Spacers in zebrafish 23 bp RSS from both immunoglobulins and TCRalpha contain 7 bp motif also resembling nonamer sequence while corresponding human sequences do not contain analogous motif. RSS are recognized by RAG1 protein, which also has specific features in teleost suggesting co-evolution of RAG1 with corresponding RSS. PMID- 15829314 TI - Relationship between Bet v 1 and Bet v 2 specific IgE and food allergy in children with grass pollen respiratory allergy. AB - Grass allergy is the most common pollinosis in Northern Italy. Some patients with grass allergy show polysensitization against other pollens and plant-derived foods. In these patients oral allergic syndrome (OAS) is frequently associated. To evaluate the correlation between food allergy or food sensitization and specific IgE against panallergens such as Bet v 1 and Bet v 2, we studied 56 children (mean age: 8 years 5 months) suffering from respiratory allergy due to grass pollens were enrolled. Specific IgE against the most important food, inhalant allergen and Bet v 1, Bet v 2 were performed by ImmunoCAP technology (UniCAP 1000, Pharmacia Diagnostics, Uppsala, Sweden). We found 14 children (25%) sensitized to Bet v 1 and 13 (23%) to Bet v 2; in 24 cases (42.3%) a sensitization to at least one of the 2 panallergens was observed. Five of the 14 cases (36%) sensitized to Bet v 1 showed food allergy and 8 (57%) food sensitization; 6 (46%) of the 13 children sensitized to Bet v 2 showed food allergy and 7 (54%) food sensitization; only one case of Bet v 1 specific IgE without food allergy or sensitization was seen. Sixteen subjects (29%) showed food allergy (group A); 20 children (35.5%) multiple sensitizations to inhalant and at least one plant-derived food (group B); 20 subjects (35.5%) only inhalant allergens (group C). Sensitization to Bet v 1 (P<0.03) and Bet v 2 (P<0.009) is from a statistical point of view significantly higher in groups A and B than in group C. In the 16 patients with food allergy hazelnut was the major triggering food (50%), followed by peanut (38%), kiwi (31%), apple and walnut (19%). Specific IgE for Bet v 1 is more associated with nuts and legumes, while Bet v 2 is more related to fresh fruit and vegetables. In conclusion grass pollinosis is frequently associated with polysensitization to other pollen and food allergens. Bet v 1 and Bet v 2 specific IgE are significantly higher in these patients than in patients with grass monosensitization, and this sensitization may be considered a possible risk factors to evolve later into food allergy. Among the offending foods, legumes and the nut group are mostly related to Bet v 1, while vegetables and fresh fruits to Bet v 2. PMID- 15829315 TI - Murine monoclonal antibody which can distinguish cystatins SA1 and SA2. AB - To develop a diagnostic trial enabling the selective examination for a target cystatin in human body fluids, we attempted to prepare monoclonal antibodies against human cystatin SA1 (originally cystatin SA) and its variant form (cystatin SA2). BALB/c mice were immunized with recombinant (r-) cystatins SA1 and SA2. Two monoclonal antibodies designated Cys3F11 and Cys2E5 were selected. By ELISA analyses, the Cys2E5 was shown to react with r-cystatin SA2 but also somewhat with r-cystatin SA1 (22% cross-reactivity) and with plasma cystatin C (18% cross-reactivity), indicating a high specificity for cystatin SA2. The Cys3F11 reacted not only with r-cystatin SA1 but also with r-cystatin SA2 (89% cross-reactivity) and plasma cystatin C (47% cross-reactivity). This finding was further emphasized by immunoblotting of human submandibular-sublingual saliva samples. ELISA additivity test suggests that the two monoclonal antibodies bind to distinct epitopes. In conclusion, we have succeeded in producing two antibodies that discriminate the structural differences between salivary cystatins S and SN, which share more than 90% identity in amino acid sequence with cystatin SA. PMID- 15829316 TI - Surgical management of malignant pleural mesothelioma: a systematic review and evidence summary. AB - An evidence summary was developed for the surgical management of adult patients with diffuse or localized malignant pleural mesothelioma. This evidence summary is based on a systematic search and review of the literature published between 1985 and February 2004. Relevant studies were identified, according to pre determined criteria by the authors and methodologists. No randomized controlled trials comparing pleurectomy (PL) with extrapleural pneumonectomy (EPP) or comparing surgery with an alternative treatment were identified. Four comparative studies, seven non-comparative prospective studies, and 16 retrospective case series were identified that looked at PL, or EPP, or PL and EPP. Trial results were not pooled due to the heterogeneity of the treatments in the trials and the fact that no trials were randomized and none were designed to directly compare different treatments. External feedback was obtained from Ontario clinicians, and the provincial Lung Cancer Disease Site Group approved the review. CONCLUSIONS: The role of surgery in the management of malignant pleural mesothelioma cannot be precisely defined as the lack of randomized controlled clinical trials makes it impossible to determine whether the use of EPP or PL improves survival or effectively palliates the symptoms of the disease. Future studies of the role of surgery in the treatment of mesothelioma should include evaluations of quality of life. PMID- 15829317 TI - Cost-effectiveness analysis of screening for lung cancer with low dose spiral CT (computed tomography) in the Australian setting. AB - INTRODUCTION: Low dose spiral computed tomography (CT) is a sensitive screening tool for lung cancer that is currently being evaluated in both non-randomised studies and randomised controlled trials. METHODS: We conducted a quantitative decision analysis using a Markov model to determine whether, in the Australian setting, offering spiral CT screening for lung cancer to high risk individuals would be cost-effective compared with current practice. This exploratory analysis was undertaken predominantly from the perspective of the government as third party funder. In the base-case analysis, the costs and health outcomes (life years saved and quality-adjusted life years) were calculated in a hypothetical cohort of 10,000 male current smokers for two alternatives: (1) screen for lung cancer with annual CT for 5 years starting at age 60 year and treat those diagnosed with cancer or (2) no screening and treat only those who present with symptomatic cancer. RESULTS: For male smokers aged 60-64 years, with an annual incidence of lung cancer of 552 per 100,000, the incremental cost-effectiveness ratio was 57,325 dollars per life-year saved and 105,090 dollars per QALY saved. For females aged 60-64 years with the same annual incidence of lung cancer, the cost-effectiveness ratio was 51,001 dollars per life-year saved and 88,583 dollars per QALY saved. The model was used to examine the relationship between efficacy in terms of the expected reduction in lung cancer mortality at 7 years and cost-effectiveness. In the base-case analysis lung cancer mortality was reduced by 27% and all cause mortality by 2.1%. Changes in the estimated proportion of stage I cancers detected by screening had the greatest impact on the efficacy of the intervention and the cost-effectiveness. The results were also sensitive to assumptions about the test performance characteristics of CT scanning, the proportion of lung cancer cases overdiagnosed by screening, intervention rates for benign disease, the discount rate, the cost of CT, the quality of life in individuals with early stage screen-detected cancer and disutility associated with false positive diagnoses. Given current knowledge and practice, even under favourable assumptions, reductions in lung cancer mortality of less than 20% are unlikely to be cost-effective, using a value of 50,000 dollars per life-year saved as the threshold to define a "cost-effective" intervention. CONCLUSION: The most feasible scenario under which CT screening for lung cancer could be cost-effective would be if very high-risk individuals are targeted and screening is either highly effective or CT screening costs fall substantially. PMID- 15829318 TI - Polymorphisms in genes involved in xenobiotic metabolism and lung cancer risk under the age of 60 years. A pooled study of lung cancer patients in Denmark and Norway. AB - The genetic susceptibility hypothesis has been used to explain why only a minority of smokers develop lung cancer. Only few studies have studied the role of polymorphisms in phase-I and II metabolizing genes, among young lung cancer patients. We have pooled the individual data of three studies from Denmark and Norway, including 320 patients diagnosed with non-small cell lung cancer at age 59 or below, and 618 age and gender matched controls. A questionnaire was used to determine relevant demographic and lifestyle characteristics, and polymorphisms in following genotypes were determined GSTM1, GSTM3, GSTP1, GSTT1, GPX1, MPO, NQO1 and NAT2. Based on the literature, the alleles of the genotypes were categorised as high- or low-risk alleles. No individual effect of the genotypes was found on the risk of lung cancer. Given a smoking exposure, the presence of high-risk alleles (or phenotypes) was generally found to increase the risk of lung cancer, although the effect modification did not reach statistical significance. A pattern of stronger protective effect was observed in carriers of more than one allele associated with lower risk of lung cancer, and a higher risk of lung cancer in carriers of one or more alleles associated with higher risk of lung cancer, but the results did not reach statistical significance. The effect modification was generally strongest at lower levels of smoking. PMID- 15829319 TI - Adenovirus-uteroglobin suppresses COX-2 expression via inhibition of NF-kappaB activity in lung cancer cells. AB - Uteroglobin (UG, Clara cell secretory protein) is a steroid inducible, multifunctional protein that is secreted by the mucosal epithelia. UG has anti proliferative and anti-metastatic effects in cancer cells. COX-2, which catalyzes the first step in the synthesis of prostanoids, has been shown to be overexpressed in tumors. This study investigated the effect of UG on the inhibition of COX-2 expression in lung cancer cells. The level of the COX-2 protein and its mRNA were decreased by UG, as demonstrated by Western blot and the RT-PCR, respectively. The EIA shows that UG suppressed PGE2 synthesis. Western blot showed that the NF-kappaB nuclear translocation was inhibited by the transduction of UG. In addition, an EMSA demonstrated the inhibition of the NF kappaB-DNA binding by UG. The luciferase assay showed that UG also inhibited the NF-kappaB-mediated transcription activity. Furthermore, transfection of the lung cancer cell lines with the COX-2 reporter gene constructs demonstrated that the transcription of COX-2 gene was suppressed by UG. These results show that the inhibition of COX-2 expression by UG transduction correlated with the suppression of NF-kappaB activity in the lung cancer cells. This suggests that UG have the possibility for the treatment of lung cancer. PMID- 15829320 TI - Quantitative evaluation of the apoptosis regulating genes Survivin, Bcl-2 and Bax in inflammatory and malignant pleural lesions. AB - AIM: We aimed to identify defects in the programmed cell death pathway that can be related to pleural malignant mesothelioma (MM) unresponsiveness to chemotherapy. MATERIALS AND METHODS: We quantified mRNA levels of the apoptosis regulating genes Survivin, member of the IAP family, Bcl-2 and Bax, members of the Bcl-2 family. We studied 22 non-neoplastic pleural samples, comprising normal and inflammatory tissue specimens, and 42 pleural MMs using real-time RT-PCR. RESULTS: Very low mRNA levels of each apoptotic gene were detected in all normal pleural samples. All three genes displayed increased mRNA levels in inflammatory and tumor specimens. Survivin levels in pleuritis and MMs were significantly increased (333% and 908%, respectively) compared to normal counterparts (p=0.0147 and 0.00349, respectively). Bcl-2 and Bax levels were increased in inflammatory pleural samples (394%, p=0.001 and 188%, p=ns, respectively) and in MMs (94%, p=ns and 88%, p=0.0163, respectively). The Bcl-2/Bax ratio was higher in pleuritis than in MMs, compared to normal pleurae (441%, p=ns and 22%, p=ns, respectively); the difference between Bcl-2/Bax ratio in inflammatory and neoplastic pleural samples was significant (p=0.00375). CONCLUSIONS: These results suggest that apoptotic defects in pleural MMs are linked to increased levels of Survivin, whereas variations in Bcl-2 and Bax expression appear less significant, although further studies are needed to highlight Bcl-2 family members interactions in apoptosis control. Survivin progressive accumulation from normal pleura to MM suggests this gene may be important in mesothelial cancerogenesis. Survivin overexpression may also be involved in pleural MM resistance to oncological therapies. Therefore, Survivin may represent a promising novel target for selective therapies. PMID- 15829321 TI - Detection of autoantibodies to livin and survivin in Sera from lung cancer patients. AB - Survivin and livin are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. Although human antibody responses to cancer-associated antigens have been detected, the response to livin has not yet been described in lung cancer patients. We examined prevalence of anti-livin antibodies in such patients with a specific enzyme linked immunosorbent assay (ELISA) using recombinant protein. Using a cutoff value for positivity determined as the mean absorbance +2S.D. for healthy control samples, 19 of 37 lung cancer patients (51.3%) were positive for anti-livin antibodies. Of 31 samples from the same lung cancer patients, 18 (58.1%) were positive for anti-survivin antibodies. When sera from 31 lung cancer patients were assessed simultaneously by anti-survivin and anti-livin ELISAs. Twenty-one patients (71%) were positive for survivin, livin, or both. Intensity of anti livin antibody responses did not correlate with intensity of anti-survivin responses. Like anti-survivin antibodies, anti-livin antibodies, thus, can be detected in many lung cancer patients. Testing for both antibodies together may prove useful in detecting lung cancer, but more extensive studies are needed to establish the clinical significance of anti-livin antibodies. PMID- 15829322 TI - Expression of YKL-40 by peritumoral macrophages in human small cell lung cancer. AB - YKL-40 is a 40 kDa protein with possible involvement in tissue remodeling, cell proliferation and angiogenesis. Elevated serum YKL-40 levels in patients with metastatic cancers (including small cell lung cancer (SCLC)) are associated with poor prognosis. The aim of this study was to identify the cellular source of YKL 40 in SCLC patient biopsies and in a panel of 20 human SCLC lines cultured in vitro and in vivo in nude mice. In general, the SCLC cell lines had no or very limited (human) YKL-40 expression, whereas, by RT-PCR a pronounced murine (i.e., stromal) YKL-40 expression was present in all tumors. YKL-40 mRNA transcripts were detected by in situ hybridization in 9 of 10 biopsies from SCLC patients, and in each case the signal was localized in the peritumoral stroma in cells of typical macrophage morphology (confirmed by a CD68 macrophage specific stain). No YKL-40 mRNA expression was found in the cancer cells, in macrophages infiltrating the solid tumor areas, or in non-malignant tissue. In conclusion, the predominant source of elevated serum YKL-40 in SCLC is peritumoral macrophages. PMID- 15829323 TI - Tumour CEA as predictor of better outcome in squamous cell carcinoma of the lung. AB - High levels of serum carcinoembryonic antigen (S-CEA) are considered a negative prognostic factor in non-small-cell lung cancer (NSCLC), while the prognostic value of tumour CEA (T-CEA) is unknown. We investigated the prognostic role of T CEA in radically resected early stage NSCLC. We measured preoperative S-CEA levels and T-CEA in 146 patients with stage 1-2 NSCLC, and analysed their influence on survival. In patients positive for T-CEA, 3-year survival was 80%, compared to 65% in T-CEA-negative patients (p=0.03). After stratification by histology, T-CEA positivity was prognostic of better survival in squamous cell carcinoma (SCC) (p=0.024) but not in adenocarcinomas (ADK) (p=0.87). Multiple Cox regression analysis showed that T-CEA positivity was an independent predictor of better survival in patients with early stage NSCLC (p=0.02). In SCC patients, the magnitude of the hazard ratio was confirmed even if the precision of the estimate is decreased (p=0.06). In conclusion, T-CEA expression appears to be an important prognostic factor in early stage SSC of the lung. PMID- 15829324 TI - Poor correspondence between clinical and pathologic staging in stage 1 non-small cell lung cancer: results from CALGB 9761, a prospective trial. AB - PURPOSE: A major problem with the staging system for non-small cell lung cancer (NSCLC) is clinical underestimation of the extent of disease. Many patients with clinical stage 1 disease do not retain that designation following surgical resection. Herein, we present data from Cancer and Leukemia Group B (CALGB) protocol 9761 evaluating the correspondence between clinical and pathologic analysis in early stage NSCLC. METHODS: Five hundred and two patients with suspected or biopsy-proven NSCLC classified as clinical stage 1 (T1-2, N0) by computed tomography (CT) scan or cervical mediastinoscopy were prospectively enrolled in CALGB 9761. The purpose of CALGB 9761 was to prospectively evaluate molecular markers of micrometastatic disease in stage 1 NSCLC. Enrollment occurred at 11 selected institutions within the CALGB. Patients with clinically suspected resectable early stage lung cancer were eligible for enrollment if they had no evidence of mediastinal or hilar adenopathy on CT scan or if they had CT evidence of potential N2 or N3 disease (lymph node > or =1.0 cm) but with negative mediastinoscopy. No prior chemotherapy or radiotherapy was permitted. RESULTS: Of the 502 patients felt to have clinical stage 1 NSCLC enrolled in CALGB 9761, 489 underwent resection with complete surgical staging and routine histopathologic analysis. From these 489 patients, only 422 (86.3%) turned out to have pathologically documented NSCLC. Of these 422 patients, 302 (71.6%) had pathologic stage 1 disease (173 stage 1A and 129 stage 1B). Despite clinical assessment of stage 1 disease, 59 (14%) patients had pathologic stage 2 disease, 57 (13.5%) had stage 3 disease, and four (0.9%) patients had stage 4 disease. Of the patients undergoing resection for clinical stage 1 NSCLC, 65 patients did not have NSCLC (44 had benign disease and 21 had malignancies other than NSCLC) and two additional patients had dual synchronous primary NSCLC tumors and were not eligible for the study. Overall, only 61.7% (302 of 489) of patients with suspected stage 1 NSCLC disease retained that stage and diagnosis after complete surgical staging, while 38.3% had an inaccurate pre-operative clinical stage or diagnosis. CONCLUSIONS: The results from this prospective trial demonstrate the poor predictive value of current clinical staging techniques in early stage NSCLC. These findings will serve as a benchmark for comparison of future clinical imaging modalities and other tests evaluating early stage NSCLC. PMID- 15829325 TI - Transesophageal endoscopic ultrasound/fine-needle aspiration diagnosis of a malignant adrenal gland in a patient with non-small cell lung cancer and a negative CT scan. AB - Adrenal metastasis is only seen on CT scan is less than 5% of patients with otherwise resectable NSCLS, but this diagnosis has a major impact on treatment and prognosis. We present a case of a patient with NSCLC and an adrenal metastasis, which was diagnosed by EUS/FNA of an enlarged adrenal gland, who had false-negative CT scan for adrenal metastasis. PET was not performed. Prospective studies are needed to assess the incremental yield of EUS/FNA over upper abdominal CT scan and PET for detecting left adrenal metastasis in patients with suspected or proven otherwise respectable NSCLC. PMID- 15829326 TI - Second-line chemotherapy in recurrent small cell lung cancer. Results from a crossover schedule after primary treatment with cisplatin and etoposide (EP regimen) or cyclophosphamide, epirubicin, and vincristin (CEV-regimen). AB - PURPOSE: To evaluate the benefit of crossover chemotherapy with etoposide and cisplatin (EP) versus cyclophosphamide, epirubicin, vincristine (CEV) at relapse after primary treatment with the opposite regimen in patients with small cell lung cancer (SCLC). Further, to compare the crossover group with patients not receiving chemotherapy. PATIENTS AND METHODS: Among 286 patients diagnosed with relapse after first-line chemotherapy, 120 patients received second-line chemotherapy and 166 patients received best supportive care. Fifty-six patients received EP after previous treatment with CEV, 52 received CEV after EP, and 12 patients were re-treated with the same regimen. Possible prognostic factors in the crossover group were identified at time for first-line chemotherapy and at relapse. The EP therapy comprised five courses of etoposide 100 mg/m(2) IV and cisplatin 75 mg/m(2) IV on day 1, followed by oral etoposide 200 mg/m(2) daily on day 2-4. The CEV-regimen was five courses of epirubicin 50 mg/m(2), cyclophosphamide 1000 mg/m(2), and vincristine 2 mg, all IV on day 1. RESULTS: Patients administered second-line chemotherapy lived significantly longer with median survival 5.3 months compared to 2.2 months in patients with best supportive care only (P<0.001). The best supportive care patients had significantly worse PS status and more resistant disease. The crossover treatment group was well balanced regarding possible prognostic factors prior to initial treatment and at recurrence. No difference in survival was found (P=0.71). Univariate analysis revealed PS at recurrence, objective tumour response from initial chemotherapy, disease stage at first-line, LDH-, NSE-, and ALP at first line to be significant prognostic factors for survival in the second-line setting. In a multivariate analysis, only PS at time of recurrence remained an independent prognostic factor (P<0.0001). CONCLUSION: Patients administered second-line chemotherapy had significantly longer survival than patients administered best supportive care. However, this difference can be explained by more negative prognostic factors in the best supportive care group. No survival difference between EP and CEV crossover chemotherapy was found. Multivariate analysis revealed PS at time of relapse as the only independent predictor of survival in the crossover recurrent SCLC group. PMID- 15829327 TI - Second-line for small cell lung cancer: how-to-do-it? PMID- 15829329 TI - Survival of stage IIIB/IV non-small cell lung cancer patients who received chemotherapy but did not participate in clinical trials. AB - This study was designed to compare the outcome of stage IIIB/IV non-small cell lung cancer patients who were treated with chemotherapy but did not participate in clinical trials of first-line chemotherapy with patients that had been treated with three clinical trials during this period. From October 1997 through October 1999, 132 patients (stage IIIB, 31 patients; stage IV, 101 patients) who received at least one dose of chemotherapy but did not participate in first-line chemotherapy trials were included. Response was evaluated in 132 patients. Six (4.5%) achieved a complete response and 32 (24.3%) achieved a partial response, resulting in an overall response rate of 28.8% (95% CI, 21.0-36.6). The median overall survival for all 132 patients was 11 months (95% CI, 9.5-12.5), and the median progression-free survival was 4.2 months (95% CI, 3.4-5.0). The median overall and progression-free survival for patients (N=129) who participated in one of three clinical trials during the study period was 13.5 months (95% CI, 11.2-15.8) and 5.6 months (95% CI, 4.9-6.0), respectively. There was no significant difference in overall and progression-free survival between patients who did or did not participate in clinical trials (overall survival: P=0.36; progression-free survival: P=0.57). Our data suggest that the survival of patients who received chemotherapy but did not participate in clinical trials was similar to patients participated in clinical trials. PMID- 15829328 TI - Docetaxel and cyclooxygenase-2 inhibition with celecoxib for advanced non-small cell lung cancer progressing after platinum-based chemotherapy: a multicenter phase II trial. AB - PURPOSE: To assess the overall and progression-free survival, response rate, and toxicity of combined docetaxel and celecoxib in the treatment of patients with non-small cell lung cancer progressing after initial chemotherapy for advanced disease. PATIENTS AND METHODS: Patients with non-small cell lung cancer and either measurable or evaluable disease experiencing progression after one or more platinum-based chemotherapy regimens given for advanced or metastatic disease, ECOG performance status 0-2, and adequate hematologic and biochemistry parameters were eligible for study inclusion; exclusion criteria included symptomatic brain metastases and full dose anti-coagulation. Therapy consisted of docetaxel 75 mg/m(2) every 21 days for a maximum of six cycles and celecoxib 400 mg orally twice daily commencing 7 days prior to docetaxel and continuing until disease progression. RESULTS: A total of 41 patients were enrolled of whom 39 were deemed eligible and received at least one course of docetaxel. The mean age of enrolled patients was 60.5 years (range, 44-77); 67% were men and 79% white. All but one patient had an Eastern Clinical Oncology Group (ECOG) performance status of 0 or 1. Most (72%) had been treated with a prior taxane. Overall survival was 11.3 months (95% confidence interval [CI]: 7.9, 15.7) and progression-free survival 19.6 weeks (95% CI: 13.5, 25.0). A response rate of 10.2% (95% CI: 3%, 24%) for all eligible and treated patients was found. Grade 3 or 4 neutropenia occurred in 10/39 patients (25.6%); one death due to neutropenic sepsis occurred. No grade 3 or 4 renal or hepatic toxicities were documented. CONCLUSION: Treatment with combination celecoxib and docetaxel is a safe regimen with a toxicity profile similar to that of docetaxel alone. Survival data are encouraging compared to historical controls and may prolong time to disease progression compared with single-agent docetaxel. PMID- 15829330 TI - Tumor specific expression of survivin-2B in lung cancer as a novel target of immunotherapy. AB - BACKGROUND: Survivin-2B is reported to be specifically expressed in numerous malignant tissues. Furthermore, survivin-2B includes the epitope peptide (survivin-2B(80-88)) which is capable of binding to HLA-A24. In this study, we evaluated whether survivin-2B could be a novel vaccine target against lung cancer. METHOD: (1) The differences in the survivin-2B expression between 15 sets of lung cancer tissues and normal lung tissues were investigated using RT-PCR. (2) The expression of survivin-2B was further examined in 42 lung cancer tissues, and the relationship between the expression and clinicopathologic factors was analyzed. (3) To compare the frequency of precursor CTL between survivin-2B positive and negative lung cancer patients, surivivin-2B(80-88) peptide-specific CTL were induced from regional lymph node lymphocytes (LNL) of four HLA-A24 (+) lung cancer patients, in whom two showed a positive survivin-2B expression of lung cancer while another two were negative, after stimulation with surivivin 2B(80-88). RESULTS: Survivin-2B was specifically expressed in lung cancer tissues, and was expressed in 17 of 42 lung cancer tissues (42.9%). Histologically, it was significantly more frequently expressed in squamous cell carcinoma than in adenocarcinoma (p=0.014). The frequency of precursor CTL in LNL was approximately one in 2.0 x 10(7) in patients with survivin-2B expression (-) lung cancer, however, it was one in 5.0 x 10(6) to 6.0 x 10(6) in those with survivin-2B expression (+) lung cancer. CONCLUSIONS: Survivin-2B was specifically expressed in lung cancer tissue, and found to specifically elicit a cellular immune response in lung cancer patients and therefore it may be a novel candidate for peptide vaccination. PMID- 15829331 TI - Thalidomide in patients with malignant pleural mesothelioma. AB - OBJECTIVES: The purpose of this study was to describe the experience with the anti-angiogenic agent thalidomide in the treatment of patients with a malignant pleural mesothelioma (MPM). MATERIALS AND METHODS: Patients with a histological confirmed diagnosis of malignant mesothelioma received thalidomide orally at night. Increasing doses of 100, 200 or maximally 400 mg were given till progression or unacceptable toxicity. Patients that did not show signs of progression for > or =6 months were considered responders. Neurological examination included the use a sensory nerve action potential (SNAP) test. RESULTS: Forty patients were evaluable for toxicity and efficacy. Twenty of them (50%) had received prior treatment. Thirty patients (75%) received a dose of 400 mg, 11 (37%) required dose reduction and 10 patients were confined to a dose of 200 mg or less. The major toxicity elicited by thalidomide concerned constipation (18/40 grade I and II) and two patients developed grade II neurotoxicity. A decline in SNAP test (>50%) did not seem to be related to the extent of neurological complaints. Eleven patients (27.5%) showed disease stabilization for >6 months and the median survival was 230 days (CI 130-330). CONCLUSIONS: The advised dose for future studies in these patients is 200 mg/day. At this dose level, the toxicity is mild and routine monitoring of neurological toxicity (SNAP test) can be omitted. The percentage of patients seen with prolonged disease stabilization upon thalidomide treatment warrants phase III studies in MPM. PMID- 15829332 TI - Levels of natural radionuclides in some Nigerian cereals and tubers. AB - The activities of radionuclides in the two major groups of foodstuff widely consumed in Nigeria have been determined. The 40K concentration range from 9.9+/ 3.6 Bq kg-1 to 298+/-14 Bq kg-1 (av. 130+/-8.12 Bq kg-1). (238)U concentration range from 1.47+/-0.91 Bq kg-1 to 39.5+/-9.9 Bq kg-1 (av. 11.5+/-3.86 Bq kg-1), and 232Th range from 3.50+/-1.85 Bq kg-1 to 10.5+/-2.13 Bq kg-1 (av. 6.78+/-2.13 Bq kg-1). 137Cs was not detected in any of the foodstuffs analysed. PMID- 15829333 TI - Characterization of the TE-NORM waste associated with oil and natural gas production in Abu Rudeis, Egypt. AB - The present study was conducted to characterize the Technically Enhanced Naturally Occurring Radioactive Materials (TE-NORM) waste generated from oil and gas production. The waste was characterized by means of dry screening solid fractionation, X-ray analysis (XRF and XRD) and gamma-ray spectrometry. Sediment of the TE-NORM waste was fractionated into ten fractions with particle sizes varying from less than 100 microm to more than 3 mm. The results showed that the TE-NORM waste contains mainly radionuclides of the 238U, 235U and 232Th series. The mean activity concentrations of 226Ra (of U-series), 228Ra (of Th-series) and 40K in the waste samples before fractionation (i.e. 3 mm) were found to amount to 68.9, 24 and 1.3 Bq/g (dry weight), respectively. After dry fractionation, the activity concentrations were widely distributed and enriched in certain fractions. This represented a 1.48 and 1.82-fold enrichment of 226Ra and 228Ra, respectively, in fraction F8 (2.0-2.5 mm) over those in bulk TE-NORM waste samples. The activity ratios of 238U/226Ra, 210Pb/226Ra, 223Ra/226Ra and 228Ra/224Ra were calculated and evaluated. Activity of the most hazardous radionuclide 226Ra was found to be higher than the exemption levels established by IAEA [International Atomic Energy Agency, 1994. International Basic Safety Standards for the Protection against Ionizing Radiation and for the Safety of Radiation Sources. GOV/2715/94, Vienna]. The radium equivalent activity (Ra-eq), radon (222Rn) emanation coefficient (EC) and absorbed dose rate (Dgammar) were estimated and these are further discussed. PMID- 15829334 TI - Self-calibration techniques of underwater gamma ray spectrometers. AB - In situ continuous monitoring of radioactivity in the water environment has many advantages compared to sampling and analysis techniques but a few shortcomings as well. Apart from the problems encountered in the assembly of the carrying autonomous systems, continuous operation some times alters the response function of the detectors. For example, the continuous operation of a photomultiplier tube results in a shift in the measured spectrum towards lower energies, making thus necessary the re-calibration of the detector. In this work, it is proved, that when measuring radioactivity in seawater, a photo peak around 50 keV will be always present in the measured spectrum. This peak is stable, depends only on the scattering rates of photons in seawater and, when it is detectable, can be used in conjunction with other peaks (40K and/or 208Tl) as a reference peak for the continuous calibration of the detector. PMID- 15829335 TI - 241Am, 137Cs, Sr and Pb uptake by tobacco as influenced by application of Fe chelators to soil. AB - To determine the potential for phytoextraction of 241Am and other contaminants from soil, accumulation of 241Am, 137Cs, Sr, Fe, Al, Pb, and Mg by tobacco was determined for soil applications of two concentrations of ethylenediaminetetraacetic acid (EDTA), citric acid, and ascorbic acid. In tobacco receiving EDTA at 3.1 mmol/kg of soil, 241Am content of plants averaged 15 Bq/kg (ranging up to 26 Bq/kg) while Fe concentrations became constant at 4.5 mmol/kg. Soil treatment with 18.8 mmol/kg EDTA resulted in average 241Am concentrations of 29 Bq/kg (19 times higher than controls). Uptake of Pb was similar to 241Am. In these samples, Fe increased to a maximum of almost 18 mmol/kg and 241Am content increased linearly with both Fe and Al. Plants receiving ascorbic and citric acids took up smaller quantities of 241Am, Pb, and Fe, even though these reagents were able to elute about as much Fe from the soil as EDTA. Synchrotron microbeam X-ray fluorescence (SXRF) was used to determine radial distributions of elements in roots and stems with and without EDTA treatment. SXRF maps indicate differences in behavior between Fe and Pb that are consistent with the bulk plant observations and provide insight into changes in metal content of the roots in the presence of EDTA. PMID- 15829336 TI - Radon-prone areas in the Lombard plain. AB - This paper reports the results of indoor radon measurements carried out in the Lombard plain. The aim of this study, which is based on the geological context, was to identify radon high-risk areas. The underlying geology has been established by means of the available stratigraphies giving a schematic representation of the sites in which either occurrence of gravel or silt and clay predominate with depths ranging from 0 to 50 m. Radon measurements were performed in a sample of 411 one-family houses in seven villages located in the southern area of Bergamo and Brescia. The findings indicate that when the substrate is dominated by clay, radon concentration for dwellings on the ground floor are low, whereas a strong predominance of underlying gravel mixed up in sand gives in winter months high radon flux from underground. PMID- 15829337 TI - On the influence of soil properties on the transfer of 137Cs from two soils (Chromic Luvisol and Eutric Fluvisol) to wheat and cabbage. AB - Two types of soils (Eutric Fluvisol and Chromic Luvisol) and two crops (wheat and cabbage) were investigated for determination of the transfer of 137Cs from soil to plant. Measurements were performed using gamma-spectrometry. Results for the soil characteristics, transfer factors of the radionuclides (TF), and conversion factors (CF) (cabbage/wheat) were obtained. The transfer of 137Cs was higher for Chromic Luvisol for both the plants. Statistically significant dependence of TF of 137Cs on its concentration in soil was established for cabbage. Dependence between K content in the soil and the transfer factor of 137Cs was not found due to the high concentrations of available K. Use of bioconcentration factor (BCF) (ratio between the activity concentration of a radionuclide in a reference plant to its concentration in another plant) is demonstrated and proposed for risk assessment studies. PMID- 15829338 TI - Transuranium radionuclide pollution in the waters of the La Maddalena National Marine Park. AB - Following the grounding and subsequent explosion, in October 2003, of a nuclear submarine in the waters of the La Maddalena National Marine Park, fears arose of possible radioactive leakages. However, isotopic analyses on algae showed that the gamma-ray emitting artificial radionuclides that one might expect to leak from a damaged nuclear reactor (such as U-235, I-131, Cs-137) were absent, and that U-238/U-234 activities were in equilibrium with values typical of sea water; this excluded any direct anthropogenic contamination as a result of the accident. We used alpha autoradiographic techniques to detect possible traces of transuranium radionuclides; 160 samples of algae, granites, sea urchins, gastropods, limpets, cuttlefish and jellyfish were collected from the area, as well as from other Mediterranean coastlines and the Baltic Sea. All samples were autoradiographed, and selected samples further analysed by alpha spectrometry. There were no alpha track concentrations above background levels in our control Mediterranean specimens. In the samples from the La Maddalena and Baltic areas two different track distributions were observed: --those homogeneously distributed over the surfaces examined; --groups (10 to over 500) of radially distributed alpha tracks (forming "star" bursts, or "hot spots") emanating from point sources. By comparing radionuclide activities measured by alpha spectroscopy with alpha track densities, we extrapolated Pu activities for all samples. About 74% of algae had Pu activities of less than 1 Bq/kg and 0.25 Bq/kg, 16% had accumulated Pu to levels between 1 and 2 Bq/kg, and a very few specimens had concentrations between 2 and 6 Bq/kg. Plots showed that alpha tracks and stars concentrate around the northern and eastern margins of the Rada (Basin) di Santo Stefano, sites facing the nuclear submarine base on the eastern shore of the island of Santo Stefano. What is the source of these nuclides: last century's atmospheric nuclear testing, Chernobyl or a local source? Their concentrated, extremely localised occurrence seems difficult to explain in terms of left-over worldwide nuclear pollution. A local source seems more plausible. PMID- 15829340 TI - Effects of ionising radiation on biota: do we need more regulation? PMID- 15829339 TI - Rodents as receptor species at a tritium disposal site. AB - New methods are being employed on the Department of Energy's Savannah River Site to deal with the disposal of tritium, including the irrigation of a hardwood/pine forest with tritiated water from an intercepted contaminant plume to reduce concentrations of tritium outcropping into Fourmile Branch, a tributary of the Savannah River. The use of this system has proven to be an effective means of tritium disposal. To evaluate the impact of this activity on terrestrial biota, rodent species were captured on the tritium disposal site and a control site during two trapping seasons in order to assess tritium exposure resulting from the forest irrigation. Control site mice had background levels of tritium, 0.02 Bq/mL, with disposal site mice having significantly higher tritium concentrations, mean=34.86 Bq/mL. Whole body tritium concentrations of the mice captured at the disposal site were positively correlated with tritium application and negatively correlated with precipitation at the site. PMID- 15829341 TI - On the possibility of molecular recognition of taste substances studied by Gabor analysis of oscillations. AB - A liquid membrane oscillator containing nitromethane as membrane material has been investigated. The influence of substances responsible for taste belonging to four classes (sweetness, saltiness, sourness and bitterness) on oscillation patterns of liquid membrane oscillators with cationic surfactant benzyldimethyltetradecylammonium chloride (BDMTAC) was examined. A new approach based on Gabor transformation is proposed for obtaining the power spectra of the observed oscillating signals and for establishing "fingerprints" of the investigated substances. It was shown that two-dimensional form of these power spectra might be used efficiently for this purpose. PMID- 15829342 TI - Energy of an ion crossing a low dielectric membrane: the role of dispersion self free energy. AB - The Born charging equation predicts that the permeability of a cell membrane to ions by the solubility-diffusion mechanism depends on the ionic radius and on the dielectric constant of the membrane. However, experiments, for example, on red blood cells and on lysosome membranes, show that the permeability depends strongly on the choice of salt anion in a way that cannot be accommodated by differences in ionic size. We demonstrate that one step towards understanding this ion specificity is to take account of the previously ignored dispersion self free energy of the ion. This is the quantum electrodynamic analogue of the (electrostatic) Born self-energy of an ion. We show that the dispersion self-free energy contribution can be and often is of the same order of magnitude as the Born contribution. To understand the observed specificity, it is essential to take into account of both ionic size and ionic polarizability. In parallel and to reinforce these observations, we also give simple estimates for how self-free energy changes that occur when an ion moves into the air-water interface region (which has a density profile for water molecules) can influence the surface tension of salt solutions. Consistency can be found between the Hofmeister sequences observed in ion permeation and in surface tension of electrolytes when these previously ignored self-free energies are included properly. PMID- 15829343 TI - A theoretical study on the activation of Ser70 in the acylation mechanism of cephalosporin antibiotics. AB - A computational study using some molecular modeling and quantum mechanical methods has been performed for determining the most favor activation process for Ser70 in the acylation mechanism for the cephalosporin antibiotics among the three proposed ones given in the literature. The computation is based on an X-ray structure of the B chain of the Bacillus licheniformis BS3 beta-lactamase cefoxitin complex. The position of a catalytic water involved in one of the reaction mechanism is defined using the Grid20 and InsightII programs, while that of the truncated ligand is defined using the InsightII and FirstDiscovery programs. The geometry of structures of each reaction scheme is optimized at the HF/3-21 G level of theory, and then the single point energy for each reactive species in each reaction scheme is computed at the levels of HF/6-31 + G (3df, 2p) and B3LYP/6-31 + G (3df, 2p). The effect of solvent on each reaction scheme is also studied by comparing the calculation results for each reaction scheme either in gas phase or in solution using the HF/6-31 + G (3df, 2p) level of theory. A computation using the B3LYP/6-31 + G (3df, 2p) level of theory and the Polarized Continuum Model (PCM) and by treating water as a solvent is also conducted for each activation process. It is found that, energetically, the most favor activation process for Ser70 in the acylation mechanism is the one where a proton transfer is mediated by the catalytic water and the catalytic residues Glu166 and Ser70. This agrees with those observed in an ultrahigh resolution X ray structure and a QM/MM theoretical study published recently on the same acylation process. PMID- 15829344 TI - Thermodynamics of the mechanism of the nitrogenase reaction. AB - The fixation of molecular nitrogen by nitrogenase requires a lot of energy because 16 mol of ATP are hydrolyzed per mole of nitrogen converted to ammonia. Kim and Dees determined the crystallograpic structure of nitrogenase and this has led to a three-step mechanism that involves Feprotein and MoFeprotein in addition to ferredoxin. Each of these steps can be interpreted in terms of two half reactions that are connected through their transfer of electrons. Estimates can be made of the standard apparent reduction potentials of these three steps and their dependencies on pH and ionic strength. This mechanism is compared with the same type of analysis of an alternative three-step mechanism in which the hydrolysis of ATP is coupled with the reduction of molecular nitrogen, rather than the reduction of Feprotein. The problem with the first mechanism is that the second step produces 12 mol of hydrogen ions per mole of nitrogen fixed and the third step consumes 10 mol of hydrogen ions per mole of nitrogen fixed. The alternative mechanism does not have this problem. PMID- 15829345 TI - Transfer of stearic acids from albumin to polymer-grafted lipid containing membranes probed by spin-label electron spin resonance. AB - Human serum albumin (HSA) has been spin-labelled with stearic acids having the nitroxide moiety attached to the hydrocarbon chain either at the 5th or at the 16th carbon atom (n-SASL, n = 5 and 16, respectively) with respect to the carboxyl groups. Its interaction with sterically stabilized liposomes (SSL) composed of dipalmitoylphosphatidylcholine (DPPC) mixed with submicellar content of poly(ethylene glycol:2000)-grafted dipalmitoyl phosphatidylethanolamine (PEG:2000-DPPE) has been studied by conventional electron spin resonance (ESR) spectroscopy. In the absence of bilayer membranes, the ESR spectra of nitroxide stearic acids non-covalently bound to HSA are single component powder patterns, indicative of spin labels undergoing temperature dependent anisotropic motion in the slow motional regime on the conventional ESR timescale. The adsorption of HSA to DPPC bilayers results in two component ESR spectra. Indeed, superimposed to an anisotropic protein-signal appears a more isotropic signal due to the labels in the lipid environment. This accounts for the transfer of fatty acids from the protein to DPPC bilayers. Two spectral components with different rotational mobility are also singled out in the spectra of n-SASL bound to HSA when DPPC/PEG:2000-DPPE mixtures are present in the dispersion medium. The fraction, f(L)(16-SASL), of spin labels transferred from the protein to lipid/polymer-lipid lamellar membranes has been quantified performing spectral subtraction. It is found that f(L)(16-SASL) decreases on increasing the content of the polymer-lipid mixed with DPPC. It is strongly reduced in the low-density mushroom regime and levels off in the high-density brush regime of the polymer-lipid content as a result of the steric stabilization exerted by the PEG-lipids. Moreover, the fraction of transferred fatty acids from HSA to SSL is dependent on the physical state of the lipid bilayers. It progressively increases with increasing the temperature from the gel to the liquid-crystalline lamellar phases of the mixed lipid/polymer-lipid membranes, although such a dependence is much weaker in the brush regime. PMID- 15829346 TI - Resonance Raman spectroscopy of carotenoids in Photosystem I particles. AB - Low-temperature resonance Raman (RR) spectroscopy was used for the first time to study the spectral properties, binding sites and composition of major carotenoids in spinach Photosystem I (PSI) particles. Excitation was provided by an argon ion laser at 457.9, 476.5, 488, 496.5, 502 and 514.5 nm. Raman spectra contained the four known groups of bands characteristic for carotenoids (called from nu(1) to nu4). Upon 514.5, 496.5 and 476.5 nm excitations, the nu(1)-nu(3) frequencies coincided with those established for lutein. Spectrum upon 502-nm excitation could be assigned to originate from violaxanthin, at 488 nm to 9-cis neoxanthin, and at 457.9 nm to beta-carotene and 9-cis neoxanthin. The overall configuration and composition of these bound carotenoid molecules in Photosystem I particles were compared with the composition of pigment extracts from the same PSI particles dissolved in pyridine, as well as to configuration in the main chlorophyll a/b light-harvesting protein complex of photosystem II. The absorption transitions for lutein, violaxanthin and 9-cis neoxanthin in spinach photosystem I particles are characterized, and the binding sites of lutein and neoxanthin are discussed. Resonance Raman data suggest that beta-carotene molecules are also present in all-trans and, probably, in 9-cis configurations. PMID- 15829347 TI - Structural studies of the biomineralized species of calcified pancreatic stones in patients suffering from chronic pancreatitis. AB - The pancreatic stones (Pancreatic calculi) collected from patients suffering from chronic calcific pancreatitis were studied in a view to explore the constituents involved in the calcification. The calcified stones were characterized by XRD, EPR and IR spectroscopic techniques. The detailed studies indicate that these stones consist of calcium carbonate as a major component, besides phosphates and other protein content. The presence of aragonite phases in the biomineralized stones is also discussed. The EPR spectra gave an evidence of the presence of traces of manganese in different oxidation states, which is used as one of the EPR probes in the present work. The samples were sintered at different temperatures to remove all the organic matter, and their EPR spectra have been studied to obtain detailed information regarding the changes in the symmetry of these stone samples. The X-irradiated sample was also characterized by EPR and the resonance signals are attributed to freely rotating CO(2)(-) radicals. The infrared spectrum reveals the presence of many organic bands corresponding to the protein amides. PMID- 15829348 TI - Improvements in technical assessment and protocol for EPR evaluation of magnetic fields effects on a radical pair reaction. AB - The effects of either static or pulsed magnetic fields on the reaction rate of Fremy's salt-ascorbic acid were studied directly by EPR spectroscopy. Radical pair mechanism (RPM) accounts for the magnetic field effects, but the expected amounts are so small that they need to be observed with particular care with EPR technique. The method is based on the resolution of a pair of EPR signals by the addition of a stationary field gradient, where the signals are coming from the exposed and control capillary sample. To this purpose, a suitable device for the gradient generation was used. Others improvements were the strictly keeping of the same boundary temperature condition in the capillary pairs, obtained by a refrigerating system controlled by a thermocouple, and the use of a pair of Helmholtz coils to generate an external high homogeneous magnetic field. By this experimental set up, we found that the magnetic field induce the decrease of the studied radical reaction rate. This EPR approach is a significant alternative to the spectrophotometric one. Moreover, it offers the advantage to detect both the radicals and/or intermediates involved in the reaction. PMID- 15829349 TI - Thermodynamics of alpha-lactalbumin-DL-alpha-dipalmitoylphosphatidylcholine interactions and effect of the antioxidant nicotinamide on these interactions. AB - Differential scanning calorimetry has been used to understand the thermodynamics of the interactions of dl-alpha-dipalmitoylphosphatidylcholine (DPPC) with alpha lactalbumin and the effect of the antioxidant nicotinamide on these interactions. Nicotinamide decreases the thermal transition temperature of both the lipid and the protein at high concentrations. The thermal unfolding transitions of the protein were two state and calorimetrically reversible. There was no significant change in the shape and thermodynamic parameters accompanying the lipid endotherms, suggesting that nicotinamide did not penetrate the lipid bilayer. The thermal unfoldings of alpha-lactalbumin in the presence of DPPC as cosolute also adhered to two-state reversible mechanism. The changes in the thermodynamic parameters accompanying the thermal transitions were small, indicating no significant interaction of alpha-lactalbumin with DPPC. The changes in the thermodynamic parameters indicate that the lipid bilayer organization, as well as the partitioning of the extrinsic protein alpha-lactalbumin into the bilayer, is not affected in the entire studied concentration range of the lipid. It is observed that the presence of increasing concentration of nicotinamide (as high as 1.0 mol dm(-3)) in the lipid-protein mixture does not affect its partitioning into the lipid bilayer, although nicotinamide preferentially interacts with alpha lactalbumin. The change in the effect of nicotinamide on lipid transition temperature in the mixture and literature report suggests that nicotinamide may be forming a hydrogen-bonded complex with the protein through its amide functionality. The surface tension data of aqueous nicotinamide in combination with the thermal denaturation results of protein in presence of nicotinamide confirmed that surface tension effect does not have any significant contribution to the effect of nicotinamide on protein. PMID- 15829350 TI - Membrane destabilizing properties of cell-penetrating peptides. AB - Although cell-penetrating peptides (CPPs), also denoted protein transduction domains (PTDs), have been widely used for intracellular delivery of large and hydrophilic molecules, the mechanism of uptake is still poorly understood. In a recent live cell study of the uptake of penetratin and tryptophan-containing analogues of Tat(48-60) and oligoarginine, denoted TatP59W, TatLysP59W and R(7)W, respectively, it was found that both endocytotic and non-endocytotic uptake pathways are involved [Thoren et al., Biochem. Biophys. Res. Commun. 307 (2003) 100-107]. Non-endocytotic uptake was only observed for the arginine-rich peptides TatP59W and R(7)W. In this paper, the interactions of penetratin, R(7)W, TatP59W and TatLysP59W with phospholipid vesicles are compared in the search for an understanding of the mechanisms for cellular uptake. While R(7)W, TatP59W and TatLysP59W are found to promote vesicle fusion, indicated by mixing of membrane components, penetratin merely induces vesicle aggregation. Studies of the leakage from dye-loaded vesicles indicate that none of the peptides forms membrane pores and that vesicle fusion is not accompanied by leakage of the aqueous contents of the vesicles. These observations are important for a proper interpretation of future experiments on the interactions of these peptides with model membranes. We suggest that the discovered variations in propensity to destabilize phospholipid bilayers between the peptides investigated, in some cases sufficient to induce fusion, may be related to their different cellular uptake properties. PMID- 15829351 TI - Protein topology affects the appearance of intermediates during the folding of proteins with a flavodoxin-like fold. AB - The topology of a native protein influences the rate with which it is formed, but does topology affect the appearance of folding intermediates and their specific role in kinetic folding as well? This question is addressed by comparing the folding data recently obtained on apoflavodoxin from Azotobacter vinelandii with those available on all three other alpha-beta parallel proteins the kinetic folding mechanism of which has been studied, i.e. Anabaena apoflavodoxin, Fusarium solani pisi cutinase and CheY. Two kinetic folding intermediates, one on pathway and the other off-pathway, seem to be present during the folding of proteins with an alpha-beta parallel, also called flavodoxin-like, topology. The on-pathway intermediate lies on a direct route from the unfolded to the native state of the protein involved. The off-pathway intermediate needs to unfold to allow the production of native protein. Available simulation data of the folding of CheY show the involvement of two intermediates with characteristics that resemble those of the two intermediates experimentally observed. Apparently, protein topology governs the appearance and kinetic roles of protein folding intermediates during the folding of proteins that have a flavodoxin-like fold. PMID- 15829352 TI - Prediction of the mutation-induced change in thermodynamic stabilities of membrane proteins from free energy simulations. AB - Comparative protein structure modeling and free energy perturbation simulation have been applied in a consecutive manner to investigate the mutation-induced stabilization of membrane proteins (MPs) in aqueous solution without knowledge of their three-dimensional structures. The calculated difference in protein solvation free energy between the wild type and a mutant compares well with their relative thermodynamic stabilities in solution. For monomeric MPs, a mutant reveals a higher stability than the wild type if the calculated solvation free energy indicates a favorable change. On the contrary, for oligomeric MPs the stability of a mutant increases as the solvation free energy of a mutated monomer becomes less favorable, indicating that the oligomeric MP mutant would be stabilized in solution due to the reduced desolvation cost for oligomerization. The present computational strategy is expected to find its way as a useful tool for assessing the relative stability of a mutant MP with respect to its wild type in solution. PMID- 15829353 TI - Lysozyme effect on structural state of model membranes as revealed by pyrene excimerization studies. AB - Steady-state measurements of pyrene fluorescence in the model bilayer membranes composed of phosphatidylcholine (PC) and its mixtures with cardiolipin (CL) have been performed to gain insight into the effect of lysozyme on molecular organization of lipid bilayer. Analysis of vibronic structure of the probe emission spectra revealed no changes in transverse distribution of pyrene monomers on varying CL contents or increasing the extent of lysozyme binding to liposomes. Excimer-to-monomer fluorescence intensity ratio has been found to reduce on lysozyme association with lipids. The magnitude of this effect increased with increasing CL content from 0 to 40 mol%. These results have been interpreted as indicating decrease in the membrane free volume on formation of both electrostatic and hydrophobic protein-lipid contacts. PMID- 15829354 TI - Temperature effect on the structural stability, similarity, and reversibility of human serum albumin in different states. AB - In order to investigate the thermal stability of human serum albumin (HAS) in three different states (aqueous solution, cast film, and solid powder), Fourier transform infrared (FTIR) spectroscopy was applied to determine the protein secondary structural changes of these HSA samples under non-isothermal or isothermal condition. The structural similarity of HSA before and after thermal treatment was also studied to estimate the thermo-reversible property of the HSA in these different states. The results indicate that with the increase of temperature, the maximum peaks at 1652 and 1547 cm(-1) (alpha-helix) shifted to 1647 and 1542 cm(-1) (random coil), respectively. An additional peak at 1620 cm( 1) assigned to intermolecular beta-sheet structure clearly appeared with temperature. The alpha-helix content was found to be reduced in favor of the formation of intermolecular hydrogen-bonded antiparallel beta-sheet structure beyond 60 degrees C in the heating process. From the data of structural similarity, HSA sample whether in solid powder or cast film form exhibited a better thermo-reversible property than HSA in aqueous solution even heating to 200 degrees C. PMID- 15829355 TI - Thermodynamics of stoichiometric biochemical networks in living systems far from equilibrium. AB - The principles of thermodynamics apply to both equilibrium and nonequilibrium biochemical systems. The mathematical machinery of the classic thermodynamics, however, mainly applies to systems in equilibrium. We introduce a thermodynamic formalism for the study of metabolic biochemical reaction (open, nonlinear) networks in both time-dependent and time-independent nonequilibrium states. Classical concepts in equilibrium thermodynamics-enthalpy, entropy, and Gibbs free energy of biochemical reaction systems-are generalized to nonequilibrium settings. Chemical motive force, heat dissipation rate, and entropy production (creation) rate, key concepts in nonequilibrium systems, are introduced. Dynamic equations for the thermodynamic quantities are presented in terms of the key observables of a biochemical network: stoichiometric matrix Q, reaction fluxes J, and chemical potentials of species mu without evoking empirical rate laws. Energy conservation and the Second Law are established for steady-state and dynamic biochemical networks. The theory provides the physiochemical basis for analyzing large-scale metabolic networks in living organisms. PMID- 15829356 TI - Parelectric spectroscopy of drug-carrier-systems--distribution of carrier masses or activation energies. AB - The answer of a high-frequency electromagnetic wave to a sample as termination of an open-ended coaxial line gives the mobility and the density of permanent electric dipole moments in the substance under test. As long as these dipoles are attached to carrier molecules of well defined masses, both parameters can be extracted from the reflected wave in a quick manner giving unambiguous results. The corresponding algorithm has been applied to solid lipid nanoparticles with glucocorticoid molecules attached to or incorporated in the carrier molecules. The results from measurements in the frequency region (0.1-100) MHz have recently been published. As soon as we have to envisage a distribution in carrier masses and/or in activation energies of the attached molecules, we have to apply a more sophisticated evaluation algorithm. The need for a more generalised algorithm is clear as well, when we have to deal with more than one dipole-carrying constituent in the samples. All these evaluation algorithms shall be presented together with the mathematical basis in a short but exact form. PMID- 15829357 TI - Transcriptional activation mechanisms of the PRM promoter of lambda phage. AB - We investigate the transcriptional activity associated with the P(RM) promoter of lambda phage. The probability for formation of a transcriptionally active (open) RNA polymerase-DNA complex is calculated by means of an equilibrium statistical mechanical model. In particular, we study two different models of the transcriptional activation mechanism when the O(R)2 site is occupied by a CI dimer (typical for a lysogen) compared to the situation when O(R)2 is vacant: (1) transcription rate increases (wild-type mechanism) or (2) RNA polymerase becomes stronger (or weaker) bound to DNA (mutant mechanism). By applying experimental determined protein-DNA binding energies we show that these two mechanisms exhibit different characteristics when we study the activity versus CI concentration. We also show that our model may be fitted to in vivo activity data satisfactorily despite that the activated transcription rate is significantly reduced compared to the wild-type value. The model is consistent with experimental determined activities based upon mutations of CI and RNA polymerase. PMID- 15829359 TI - A linearization method for low catalytic activity enzyme kinetic analysis. AB - A kinetic analysis was made and a linear plot based on the general rate equation derived by Laidler [Can. J. Chem. 33, 1614-1624] is proposed. This linearization method allows determining the kinetic parameters (K(m), k(cat)) and [E](0) for enzymes with low catalytic activity. The method was applied to chloroperoxidase from Caldariomyces fumago [EC 1.11.1.10], whose kinetic parameters K(m)(app), k(cat)(app), and [E](0) with monochlorodimedone as substrate, were obtained by using the linearization plot and the V(max) value (calculated by Eadie-Hofstee plot). This plot could also be useful to the study of abenzyme kinetics provided the concentration of the latter is either higher or equal than K(m) value. PMID- 15829358 TI - Kinetic studies of iron deposition catalyzed by recombinant human liver heavy and light ferritins and Azotobacter vinelandii bacterioferritin using O2 and H2O2 as oxidants. AB - The discrepancy between predicted and measured H(2)O(2) formation during iron deposition with recombinant heavy human liver ferritin (rHF) was attributed to reaction with the iron protein complex [Biochemistry 40 (2001) 10832-10838]. This proposal was examined by stopped-flow kinetic studies and analysis for H(2)O(2) production using (1) rHF, and Azotobacter vinelandii bacterial ferritin (AvBF), each containing 24 identical subunits with ferroxidase centers; (2) site-altered rHF mutants with functional and dysfunctional ferroxidase centers; and (3) recombinant human liver light ferritin (rLF), containing no ferroxidase center. For rHF, nearly identical pseudo-first-order rate constants of 0.18 s(-1) at pH 7.5 were measured for Fe(2+) oxidation by both O(2) and H(2)O(2), but for rLF, the rate with O(2) was 200-fold slower than that for H(2)O(2) (k = 0.22 s(-1)). A Fe(2+)/O(2) stoichiometry near 2.4 was measured for rHF and its site altered forms, suggesting formation of H(2)O(2). Direct measurements revealed no H(2)O(2) free in solution 0.5-10 min after all Fe(2+) was oxidized at pH 6.5 or 7.5. These results are consistent with initial H(2)O(2) formation, which rapidly reacts in a secondary reaction with unidentified solution components. Using measured rate constants for rHF, simulations showed that steady-state H(2)O(2) concentrations peaked at 14 muM at approximately 600 ms and decreased to zero at 10-30 s. rLF did not produce measurable H(2)O(2) but apparently conducted the secondary reaction with H(2)O(2). Fe(2+)/O(2) values of 4.0 were measured for AvBF. Stopped flow measurements with AvBF showed that both H(2)O(2) and O(2) react at the same rate (k = 0.34 s(-1)), that is faster than the reactions with rHF. Simulations suggest that AvBF reduces O(2) directly to H(2)O without intermediate H(2)O(2) formation. PMID- 15829360 TI - Autoxidation studies of extracellular hemoglobin of Glossoscolex paulistus at pH 9: cyanide and hydroxyl effect. AB - The complex oligomeric assembly of the hemoglobin subunits may influence the autoxidation rate. To understand this relation, the rate of autoxidation was studied at pH 9.0, where the Glossoscolex paulistus Hemoglobin (GpHb) dissociates. At alkaline pH, this hemoglobin is dissociated into monomers, trimers and tetramers, allowing the study of the integral protein and monomer subunit autoxidation on independent experiments. The autoxidation rate was evaluated in the presence and absence of cyanide (CN(-)), a strong field ligand to the ferric ion. The oxidation kinetic was monitored using the UV-vis absorption at 415 nm, and resulted in: i) bi-exponential kinetics for the whole hemoglobin (indicating a fast and a slow oxidative process) and ii) mono exponential for the monomer (indicating a single process). To understand the specific characteristics of each autoxidation process, Arrhenius plots allowed the determination of the activation energy. The experimental results indicate for the whole hemoglobin in the absence of CN(-) an activation energy of 150 +/- 10 kJ mol(-1) for the fast and the slow processes. Under the same conditions the monomer displayed an activation energy of 160 +/- 10 kJ mol(-1), very close to the value obtained for the integral protein. The pseudo-second order rate constant for the whole protein autoxidation by CN(-) showed two different behaviors characterized by a rate constant k(CN1)' = 0.11 +/- 0.02 s(-1) mol(-1) L for CN(-) concentrations lower than 0.012 mol L(-1); and k(CN1)" = 0.76 +/- 0.04 s(-1) mol(-1) L at higher concentrations for the fast process, while the slow process remain constant with k(CN2) = 0.033 +/- 0.002 s(-1) mol(-1) L. The monomer has a characteristic rate constant of 0.041 +/- 0.002 s(-1) mol(-1) L for all cyanide concentrations. Comparing the results for the slow process of the whole hemoglobin and the oxidation of the monomer, it is possible to infer that the slow process has a strong contribution of the monomer in the whole hemoglobin kinetic. Moreover, as disulfide linkers sustain the trimer assembly, cooperativity may explain the higher kinetic constant for this subunit. PMID- 15829361 TI - Sequestering of p53 into DNA-protein filaments revealed by electron microscopy. AB - Using electron microscopy, we analyzed the interaction of bacterially expressed full-length p53, p53(1-393), and its C-terminal fragment, p53(320-393), with long (approximately 3000 bp) dsDNA in linear and supercoiled (|DeltaLk| approximately 4-6) forms containing or lacking the p53 recognition sequence (p53CON). The main structural feature of the complexes formed by either protein was a DNA-protein filament, in which two DNA duplexes are linked (synapsed) via bound protein tetramers. The efficiency of the synapse, reflected in its length and the fraction of molecules exhibiting DNA-protein filaments, was significantly modulated by the molecular form of the protein and the topological state of the DNA. With linear DNA, the synapse yield promoted by the C-terminus fragment was very low, but the full-length protein was effective in linking noncontiguous duplexes, leading to the formation of intramolecular loops constrained at their bases by short regions of synapsed DNA duplexes. When the linear DNA contained p53CON, regions of preferential sequence, i.e., encompassing p53CON and probably p53CON-like sequences, were predominantly synapsed, indicating a sequence specificity of the p53 core domain. With scDNA, the synapse yield was significantly higher compared to the linear counterparts and was weakly dependent on the sign of superhelicity and presence or absence of p53CON. However, the full length protein was more effective in promoting DNA synapses compared to the C terminal fragment. The overall structure of the DNA-protein filaments was apparently similar for either protein form, although the apparent width differed slightly (approximately 7-9 nm and approximately 10-12 nm for p53(320-393) and p53(1-393), respectively). No distortion of the DNA helices involved in the synapse was found. We conclude that the structural similarity of DNA-protein filaments observed for both proteins is attributable mainly to the C-terminus, and that the yield is dictated by the specific and possibly nonspecific interactions of the core domain in combination with DNA topology. Possible implications for the sequestering of p53 in DNA-protein filaments are discussed. PMID- 15829362 TI - Effects of anthropogenic fire history on savanna vegetation in northeastern Namibia. AB - Anthropogenic fires in Africa are an ancient form of environmental disturbance, which probably have shaped the savanna vegetation more than any other human induced disturbance. Despite anthropogenic fires having played a significant role in savanna management by herders, previous ecological research did not incorporate the traditional knowledge of anthropogenic fire history. This paper integrates ecological data and anthropogenic fire history, as reconstructed by herders, to assess landscape and regional level vegetation change in northeastern Namibia. We investigated effects of fire frequency (i.e. <5, 5-10 and >10 years) to understand changes in vegetation cover, life form species richness and savanna conditions (defined as a ratio of shrub cover to herbaceous cover). Additionally, we analysed trends in the vegetation variables between different fire histories at the landscape and regional scales. Shrub cover was negatively correlated to herbaceous cover and herbaceous species richness. The findings showed that bush cover homogenisation at landscape and regional scales may suggest that the problem of bush encroachment was widespread. Frequent fires reduced shrub cover temporarily and promoted herbaceous cover. The effects on tree cover were less dramatic. The response to fire history was scale-independent for shrub, herbaceous and tree cover, but scale-dependent for the richness of grass and tree life forms. Fire history, and not grazing pressure, improved savanna conditions. The findings emphasise the need to assess effects of anthropogenic fires on vegetation change before introducing new fire management policies in savanna ecosystems of northeastern Namibia. PMID- 15829363 TI - Land-use intensification and environmental degradation: empirical evidence from irrigated and rain-fed farms in south eastern Nigeria. AB - When evaluating the potential of irrigation for agricultural production one must consider more than the technical and financial potential and feasibility of the scheme. Issues of environmental and social sustainability must also be considered. The purpose of the paper is to develop a framework for assessing the sustainability of an irrigation scheme and apply it in the context of one such scheme in south eastern Nigeria. The framework for assessment is based on a comparison of differential changes in environmental quality with and without the scheme, and before and after the scheme. The author does this by using measured soil characteristics for irrigated and adjacent rain-fed plots of agricultural land; and socio-economic analysis of other environmental and social impacts of the irrigation scheme. Data were collected through a field survey of the selected irrigated and rain-fed farm plots, qualitative interviews with the farm owners and relevant secondary sources. The analyses find that the soils of the irrigated farms have been significantly degraded more than those of the rain-fed farms to the extent that precludes sustainable practice of arable agriculture on irrigated land in the study area. About 9% of the soils of the irrigated land have been degraded to the extent that they are no longer suitable for arable agriculture. Analyses of the qualitative interviews also find other bio-physical, social and economic impacts that significantly constrain long-term sustainability of arable agriculture in the study area. PMID- 15829364 TI - A river water quality model integrated with a web-based geographic information system. AB - Scientists often use mathematical models to assess river water quality. However, the application of the models in environmental management and risk assessment is quite limited because of the difficulty of preparing input data and interpreting model output. This paper presents a study that links ArcIMS, a Web-based Geographic Information System (GIS) software to ROUT, a national and regional scale river model which evolved from the US Environmental Protection Agency's Water Use Improvement and Impairment Model, to create a WWW-GIS-based river simulation model called GIS-ROUT. GIS-ROUT is used to predict chemical concentrations in perennially flowing rivers throughout the continental United States that receive discharges from more than 10,000 publicly owned wastewater treatment plants (WWTPs). The WWTP chemical loadings are calculated from per capita per day disposal of product ingredients and the population served by each plant. Each WWTP, containing data on treatment type and influent and effluent flows, is spatially associated with a specific receiving river segment. Based on user defined treatment-type removal rates for a particular chemical, an effluent concentration for each WWTP is calculated and used as input to the river model. Over 360,000 km of rivers are modeled, incorporating dilution and first order loss of the chemical in each river segment. The integration of spatial data, GIS, the WWW, and modeling in GIS-ROUT makes it possible to organize and analyze data spatially, and view results on interactive maps as well as tables and distribution charts. The integration allows scientists and managers in different locations to coordinate and share their estimations for environmental exposure and risk assessments. PMID- 15829365 TI - Simplified LCA and matrix methods in identifying the environmental aspects of a product system. AB - In order to effectively integrate environmental attributes into the product design and development processes, it is crucial to identify the significant environmental aspects related to a product system within a relatively short period of time. In this study, the usefulness of life cycle assessment (LCA) and a matrix method as tools for identifying the key environmental issues of a product system were examined. For this, a simplified LCA (SLCA) method that can be applied to Electrical and Electronic Equipment (EEE) was developed to efficiently identify their significant environmental aspects for eco-design, since a full scale LCA study is usually very detailed, expensive and time consuming. The environmentally responsible product assessment (ERPA) method, which is one of the matrix methods, was also analyzed. Then, the usefulness of each method in eco-design processes was evaluated and compared using the case studies of the cellular phone and vacuum cleaner systems. It was found that the SLCA and the ERPA methods provided different information but they complemented each other to some extent. The SLCA method generated more information on the inherent environmental characteristics of a product system so that it might be useful for new design/eco-innovation when developing a completely new product or method where environmental considerations play a major role from the beginning. On the other hand, the ERPA method gave more information on the potential for improving a product so that it could be effectively used in eco-redesign which intends to alleviate environmental impacts of an existing product or process. PMID- 15829366 TI - A decade of recreation ratings for six silviculture treatments in Western Oregon. AB - Managed forests are increasingly being used for recreation. As a result, foresters may be expected to tailor silvicultural treatments to accommodate specific recreation preferences. To better understand changes in hiking and camping quality in the years following a harvest, six sites on the Oregon State University's research forest were evaluated annually for 11 years. Multiple comparison and regression analyses were used to describe the data. Results show that recreation ratings generally improved over time; recreation ratings were related to but different from scenic ratings; and there were differences among recreation activities. Although several studies have previously examined recreation quality after harvest, we know of no other study that has tracked the ratings of individual harvest units through the early stages of stand regeneration. PMID- 15829367 TI - Use of questionnaires and an expert panel to judge the environmental consequences of chemical spills for the development of an environment-accident index. AB - Assessing the environmental consequences of a chemical accident is a complex task. To date, the methods used to evaluate the environmental effects of an acute release of a chemical have often been based on measurements of chemical and physical variables deemed to be important, such as the concentration of the chemical. However, a broader strategy is needed to predict the environmental consequences of potential accidents during the planning process. An Environment Accident Index (EAI), a simple tool based on such a strategy, has been developed to facilitate the consideration of a multitude of influential variables. The objectives of this study were to evaluate whether questionnaire-based expert panel's judgements could provide useful data on the environmental consequences of chemical spills, and an effective basis for further development of the EAI. As expected, the judgements did not agree perfectly, but they do give rough indications of the environmental effects, and highlight consistent trends that should be useful inputs for planning, prevention and decontamination processes. The different accidents were also judged to have caused everything from minor to very major effects in the environment, implying that a wide range of accident scenarios were represented in the material and covered by the EAI. Therefore, questionnaires and expert panel judgements can be used to collect useful data for estimating the likely environmental consequences of chemical accidents and for further development of the EAI. PMID- 15829368 TI - The use of forests for the purpose of tourism: the case of Belek Tourism Center in Turkey. AB - The main aim of this article is to examine and review the usage of forests for the purpose of tourism in Turkey with the case study of Antalya-Belek. The use and conversion of forest lands is central to tourism development. Land use change is responsible for the majority of the negative environmental impacts of tourism on natural resources. Worldwide, forests and coastal zones are converted for the construction of tourist facilities. The rapid emergence of mass tourism development in Turkey, encouraged without considering protection and sustainable use of natural and cultural resources, has resulted in serious problems in forest areas. This paper is particularly concerned with land use change in forests (or deforestation) created by the allocation of forest lands to build tourist facilities. In Belek, a tourism center located in Antalya province's coastal areas and the fastest growing destination of the country, all tourist facilities have been established in forest lands under the status of conservation forest. Today, the Belek Conservation Forest is under severe pressure from tourism. PMID- 15829369 TI - Atmospheric fate of OH initiated oxidation of terpenes. Reaction mechanism of alpha-pinene degradation and secondary organic aerosol formation. AB - This paper studies the reaction products of alpha-pinene, beta-pinene, sabinene, 3-carene and limonene with OH radicals and of alpha-pinene with ozone using FT-IR spectroscopy for measuring gas phase products and HPLC-MS-MS to measure products in the aerosol phase. These techniques were used to investigate the secondary organic aerosol (SOA) formation from the terpenes. The gas phase reaction products were all quantified using reference compounds. At low terpene concentrations (0.9-2.1 ppm), the molar yields of gas phase reaction products were: HCHO 16-92%, HCOOH 10-54% (OH source: H2O2, 6-25 ppm); HCHO 127-148%, HCOOH 4-6% (OH source: CH3ONO, 5-8 ppm). At high terpene concentrations (4.1-13.2 ppm) the results were: HCHO 9-27%, HCOOH 15-23%, CH3(CO)CH3 0-14%, CH3COOH 0-5%, nopinone 24% (only from beta-pinene oxidation), limona ketone 61% (only from limonene oxidation), pinonaldehyde was identified during alpha-pinene degradation (OH source H2O2, 23-30 ppm); HCHO 76-183%, HCOOH 12-15%, CH3(CO)CH3 0-12%, nopinone 17% (from beta-pinene oxidation), limona ketone 48% (from limonene oxidation), pinonaldehyde was identified during alpha-pinene degradation (OH source CH3ONO, 14-16 ppm). Pinic acid, pinonic acid, limonic acid, limoninic acid, 3-caric acid, 3-caronic acid and sabinic acid were identified in the aerosol phase. On the basis of these results, we propose a formation mechanism for pinonic and pinic acid in the aerosol phase explaining how degradation products could influence SOA formation and growth in the troposphere. PMID- 15829370 TI - Exploring beliefs about cancer among American Samoans: focus group findings. AB - OBJECTIVE: To evaluate answers to the following questions among American Samoans: What is cancer? What causes cancer? And what can you do to prevent cancer? DESIGN: Focus groups (four with women and four with men). SETTINGS: Pago Pago and the Manu'a islands, American Samoa; Honolulu, Hawaii; Los Angeles, California. PARTICIPANTS: 80 self-reported Samoan men and women over the age of 18 years, selected through non-probability purposive sampling with help from Samoan community-based organizations. MEASUREMENT: Qualitative content analysis of focus findings to identify themes. RESULTS: The concepts that cancer was not a Samoan illness, that failure to follow fa'aSamoa (the traditional Samoan way of life) could lead to cancer, and that a return to fa'aSamoa could prevent cancer were the prevalent themes in the focus groups. CONCLUSION: The value that Samoans place on fa'aSamoa, a traditional healthy lifestyle, provides insights into the design of future intervention programs aimed at improving cancer control in this population. PMID- 15829371 TI - Screening mammography in Minnesota cancer patients. AB - BACKGROUND: Although a decrease in mortality due to breast cancer is the most definitive measure to assess the effectiveness of screening mammography, stage at diagnosis is an interim measure. OBJECTIVES: The objectives were to (1) examine the association between screening mammography and cancer stage at diagnosis and (2) provide information about mammography utilization in an insured population diagnosed with cancer. RESEARCH DESIGN: We utilized an existing data set of linked cancer registry-claims data. We computed an odds ratio for the association between screening mammography and breast cancer stage at diagnosis and examined mammography utilization patterns. SUBJECTS: Women in Minnesota with available claims data who developed cancer in 1995. MEASURES: Mammography claims and breast cancer stage at diagnosis (grouped according to AJCC and summary stage). RESULTS: Screening mammography was associated with significantly increased odds of an early (versus late) stage cancer diagnosis, consistent with the goal of screening. Mammography utilization was generally highest in the age group 50-64 and decreased in each successive age group. CONCLUSIONS: Linkage between insurance claims and cancer registry data provides useful information not available in either data set alone. The results contribute to the cohesiveness of the evidence that mammography prevents death due to breast cancer. PMID- 15829372 TI - Hormone replacement therapy and invasive and borderline epithelial ovarian cancer risk. AB - Risk of epithelial ovarian cancer (EOC) attendant to use of hormone replacement therapy (HRT) was evaluated in a population-based case-control study of newly diagnosed EOC cases (n=256) and randomly selected population controls (n=1122). Telephone interviews were conducted to obtain information on history of HRT and several other covariates. Multivariate adjusted odds ratio (OR) and 95% confidence intervals (CI) were derived from unconditional logistic regression. The OR for ever use of HRT was 1.39 (95% CI: 1.01-1.93) compared to never use. Long-term use (>10 years) increased risk (OR: 1.62, CI: 1.05-2.50) although the trend p-value for duration of use was of only borderline significance (p=0.08). The relationship was stronger in women without hysterectomy (OR: 1.66, 95% CI: 1.14-2.41) or tubal ligation (OR: 1.56, 95% CI: 1.08-2.26). In this study, use of HRT is associated with an increased risk of EOC. PMID- 15829373 TI - Capacity building for the clinical investigation of AIDS malignancy in East Africa. AB - PURPOSE: To build capacity in the resource-poor setting to support the clinical investigation and treatment of AIDS-related malignancies in a region of the world hardest hit by the AIDS pandemic. METHODS: An initial MEDLINE database search for international collaborative partnerships dedicated to AIDS malignancies in developing countries failed to identify any leads. This search prompted us to report progress on our collaboration in this aspect of the epidemic. Building on the formal Uganda-Case Western Reserve University (Case) Research Collaboration dating back to 1987, established NIH-supported centers of research excellence at Case, and expanding activities in Kenya, scientific and training initiatives, research capital amongst our institutions are emerging to sustain a international research enterprise focused on AIDS and other viral-related malignancies. RESULTS: A platform of clinical research trials with pragmatic design has been developed to further enhance clinical care and sustain training initiatives with partners in East Africa and the United States. An oral chemotherapy feasibility trial in AIDS lymphoma is near completion; a second lymphoma trial of byrostatin and vincristine is anticipated and a feasibility trial of indinavir for endemic Kaposi's sarcoma is planned. CONCLUSIONS: In the absence of published reports of evolving international partnerships dedicated to AIDS malignancy in resource constrained settings, we feel it important for such progress on similar or related international collaborative pursuits to be published. The success of this effort is realized by the long-term international commitment of the collaborating investigators and institutions to sustain this effort in keeping with ethical and NIH standards for the conduct of research; the provision of formal training of investigators and research personnel on clinical problems our East African partners are faced with in practice and the development of pragmatic clinical trials and therapeutic intervention to facilitate technology transfer and enhance clinical practice. PMID- 15829374 TI - Diets, polymorphisms of methylenetetrahydrofolate reductase, and the susceptibility of colon cancer and rectal cancer. AB - The aim of this study was to investigate the association of environmental factors (dietary folate, methionine and drinking status) and polymorphisms in the methylenetetrahydrofolate reductase (MTHFR C677T and A1298C) gene, as well as the combination of these factors, with the risk of colon cancer and rectal cancer. A case-control study of 53 colon cancer patients, 73 rectal cancer patients and 343 healthy controls was conducted. Genotypes of C677T and A1298C polymorphisms were analyzed by PCR-RFLP. The dietary folate and methionine intakes were assessed using food-frequency questionnaires and food consumption tables. Unconditional logistic regression was applied to estimate the odds ratios (ORs) and their 95% confidence intervals (CIs). The frequency of MTHFR 677T and 1298C alleles in healthy population were 39.4 and 17.2%, respectively. After adjustment for specific variants, the MTHFR 677TT genotype showed a significantly reduced risk of colon cancer compared with the wild type (OR=0.22, 95% CI: 0.50-0.98), and 1298C allele-carrier showed an inverse association with the risk of rectal cancer compared to the wild type (OR=0.52, 95% CI: 0.28-0.98). Adequate intake of folate was a protective factor from colon cancer (OR=0.32, 95% CI: 0.12-0.88) and MTHFR C677T polymorphism showed a statistically significant effect (OR=0.25, 95% CI: 0.06-0.93), reducing the risk of colon cancer in groups that have an intake of folate exceeding 115.64ng per 1000kcal per day. This study suggests that MTHFR C677T and A1298C polymorphisms are associated with the reduced risk of colon and rectal cancers, respectively. Adequate folate intake shows an inverse association with the risk of colon cancer. There is a significant interaction between MTHFR C677T polymorphism and folate intake in reducing the risk of colon cancer. PMID- 15829375 TI - Selective sensitivity to wasabi-derived 6-(methylsulfinyl)hexyl isothiocyanate of human breast cancer and melanoma cell lines studied in vitro. AB - Recently, attention has focused on the anticancer properties of an aromatic component 6-(methylsulfinyl)hexyl isothiocyanate (6-MITC) in a typical Japanese spice, wasabi. In this paper, anticancer activity of 6-MITC in vitro was studied by using a human cancer cell (HCC) panel. 6-MITC directly affected the cells in the HCC panel and inhibited their growth in culture. The mean concentration required to inhibit 50% of control cell growth was 3.9 microM, which is a sufficiently low dosage for practical use. The suppression influenced not only the cell growth, but also the survival of these cells. The mean concentration to suppress cells to a 50% survival was 43.7 microM. The reduction activity of 6 MITC was differential, and it suppressed specific cells. These severely suppressed cell lines included breast cancer and melanoma cell lines. For example, one melanoma line was seriously damaged at a concentration of 0.3 microM of 6-MITC. Compared with other MITCs (2-MITC, 4-MITC and 8-MITC), 6-MITC showed the most effective suppression and with the most specific manner of the cells mentioned above. A "COMPARE" analysis using a computerized algorithm, which was based on the HCC database, suggested that the suppression mechanism of 6-MITC is unique and may be different from that of other known chemicals. The actual mechanism may not a simple one but may involve multiple pathways. On account of its sufficiently small size, 6-MITC is a new possible candidate for controlling cancer cells. PMID- 15829376 TI - Focal prostate basal cell layer disruptions and leukocyte infiltration are correlated events: A potential mechanism for basal cell layer disruptions and tumor invasion. AB - To assess the potential correlation between basal cell layer disruptions and leukocyte infiltration, consecutive sections of normal (n=5) and tumor (n=50) prostate tissues were double immunostained for cytokeratin 34betaE12 (CK 34betaE12) plus leukocyte common antigen, Ki-67, or proliferating cell nuclear antigen (PCNA). Of 2047 acini and ducts examined, 201 contained focal basal cell layer disruptions. Of those, 183 (91%) showed leukocyte infiltration, compared to 67 (33.3%) in 201 morphologically comparable structures with an intact basal cell layer (P<0.01). Basal cell layers adjacent to or surrounded by leukocytes were often attenuated or fragmented, and leukocytes were generally located at or near disruptions. Disrupted basal cell layers showed a markedly reduced proliferation rate, compared to their non-disrupted counterparts. Cells overlying focal basal cell layer disruptions often displayed distinct changes in the size, nuclear shape, density, and polarity, compared to those away from disruptions. A vast majority of proliferating tumor cells were located at or near basal cell layer disruptions. These findings suggest that focal basal cell layer disruptions and leukocyte infiltration are correlated events, representing a potential trigger factor for prostate tumor invasion. PMID- 15829377 TI - PTEN gene therapy induces growth inhibition and increases efficacy of chemotherapy in prostate cancer. AB - The PTEN gene, located on chromosome 10, is a phosphatase in the phosphatidylinositol 3'-kinase (PI3'K)-mediated signal transduction pathway. PTEN inhibits the activation of Akt, a serine-threonine kinase involved in proliferative metabolic and anti-apoptotic pathways, and has tumor suppressor properties. We used a PTEN adenoviral vector, Ad-MMAC, to assess the role of PTEN in the treatment of prostate cancer. Infection of Ad-MMAC in PC-3 and LNCaP prostate cancer cells (PTEN deleted, up-regulation of phosphorylated Akt) resulted in PTEN expression and significantly decreased growth compared with Ad CTR or mock infected cells. Infection of Ad-MMAC did not inhibit the growth of DU 145 cells (wild-type PTEN). Combination therapy with Ad-MMAC and doxorubicin improved the efficacy of PTEN gene therapy in PC-3 and DU-145 cells. These data demonstrate that PTEN gene therapy can effectively treat some prostate cancers that have genomic alterations in PTEN. In others, PTEN gene therapy combined with chemotherapy is more effective. PMID- 15829378 TI - Soluble Fas and Fas ligand provide new information on metastasis and response to chemotherapy in SCLC patients. AB - BACKGROUND: The Fas/Fas ligand (FasL) system is a major regulator of apoptosis. Chemotherapeutic drugs have been shown to induce Fas expression on the surface of lung cancer cells, and cancer cell apoptosis. However, this mechanism is not considered to be associated with Fas expressed on lung cancer cells. Soluble Fas and FasL concentrations are reportedly elevated in the peripheral blood of patients with lung cancer, but the roles of circulating soluble Fas and FasL in that disease have not been clarified. MATERIALS AND METHODS: We measured the circulating soluble Fas and FasL levels in 21 patients with small cell lung cancer (SCLC), and 12 healthy matched controls, in order to examine whether such ligands could provide any important information and/or reveal any new clinical features of SCLC. RESULTS: In the CR patients, the neuronal specific enolase (NSE), soluble Fas and soluble FasL concentrations were 21.26+/-3.65 ng/ml, 3.58+/-0.19 ng/ml and 0.50+/-0.15 ng/ml, while in the partial response (PR)/no change (NC)/progressive disease (PD) group of patients they were 33.96+/-7.86 ng/ml, 5.29+/-0.29 ng/ml and 0.59+/-0.07 ng/ml, respectively. The NSE, soluble Fas and soluble FasL concentrations were all elevated in the PR/NC/PD patients, however, significant differences were only seen in Fas concentration between CR and PR/NC/PD patients and CR patients and the controls (p<0.001). CONCLUSIONS: Serum soluble Fas and FasL play important roles in the proliferation and metastasis of SCLC, as well as in the cytotoxic reaction and apoptosis induced by anticancer drugs in SCLC. Further study of the mechanisms and participation of circulating soluble Fas and FasL is necessary to develop treatment strategies for SCLC. PMID- 15829379 TI - Evaluation of infections in non-small cell lung cancer patients treated with radiotherapy. AB - PURPOSE: We aim to determine infections occurring in patients with non-small cell lung cancer during radiotherapy (RT). METHODS AND MATERIALS: A total of 181 patients had been treated with thoracic radiotherapy between October 1995 and December 1999. Radiotherapy was given using 1.8-3Gray (Gy) fraction daily, five fractions a week for a total dose of 59.4Gy (30-70.2Gy). A complete history was collected retrospectively for each patient. All microbiological examinations were performed according to the routine procedures of the hospital laboratory. Numeric and categoric variables were employed such as sex, age, performance status, histology, stage, chemotherapy, usage of corticosteroids, neutropenia, surgery, hospitalization, associated diseases, smoking during treatment, package per year of cigarette smoking, dose of radiotherapy, and response rates. RESULTS: Infections developed in 84 patients (46%, 84/181) during thoracic radiotherapy. A 101 episodes of infections developed in these patients. Most patients suffered from sputum production (65%), cough (59%), auscultation findings (31%) and fever (31%). Gram-negative bacteria were the most frequently isolated pathogens in the cultures of specimens (70%, 16/23 samples). Neoadjuvant chemotherapy (OR=4.81; 95% CI, 1.57-9.12; p=0.003) and neutropenia (OR=4.25; 95% CI, 1.44-6.89; p=0.009) were found as risk factors for influencing infection based on logistic regression analyses. Package per year of cigarette smoking was found statistically significantly higher in patients with infections than patients without infections (p=0.001). A slight increase in infections, which was of borderline statistical significance (p=0.07), was observed in patients age over 70. Ciprofloxacin and clarithromycin were the most frequently used agents in treatment. Median survival was 9 months in the patients with infection and 13 months in the 97 patients without infection. Overall survival seemed to be statistically significantly better in patients without infection than patients with infection (p=0.042) calculated using Kaplan-Meier method. Based on Cox regression analyses; overall survival was not correlated to presence of infection but associated with poor performance status (5940 cGy (OR=2.06; 95% CI, 0.72-7.18; p=0.007) and the absence of response to treatment (OR=2.45; 95% CI, 0.89-14.23; p<0.001) were also found to be risk factors for survival. CONCLUSIONS: Infections are important causes of morbidity and mortality in lung cancer patients. The control of infection in these patients may improve the survival. Predisposing factors and treatment management approaches in non-small cell lung cancer should be defined carefully. PMID- 15829380 TI - Multi-probe RPA template sets to study RNA modulation and transcriptional control of BH3-only members of the Bcl-2 family. AB - The Bcl-2 family of apoptotic regulators comprises both pro- and anti-apoptotic proteins. Pro-apoptotic BH3-only members of the Bcl-2 family are subject to stringent control in normal proliferating cells, but can be activated by a variety of mechanisms in response to diverse apoptotic signals. To facilitate the simultaneous identification of BH3-only targets that are regulated at the RNA level, we have designed and assembled multi-probe ribonuclease protection assay templates and demonstrated their ability to detect multiple BH3-only transcripts in a variety of human cell lines and primary tissues. Following this, we have determined the modulation of BH3-only transcripts to a variety of apoptotic stimuli, in selected primary and transformed cell lines. Early experiments, showing that a given cell type upregulates a unique subset of BH3-only RNAs in response to different death-inducing stimuli, indicate that the multi-probe BH3 only RPA template sets could serve as useful diagnostic and analytical tools in cancer research. PMID- 15829382 TI - The pathophysiology of acute pain. AB - The method by which the body detects, interprets, and responds to painful stimulation is tantamount to an orchestral concert, with each individual instrument contributing a subtle yet important element composing the finished product. The polymodal peripheral receptors initiate unpleasant sensations that can be modulated in the dorsal horn and the anterior columns of the spinal cord before reaching the cerebral cortex. Despite similar neurological "wiring," pain sensations from the skin are not the same in character as those from visceral organs. A reaction of pain is based upon interpretation in specific cerebral centers and the resultant motor function. With the advent of functional MRI and PET scanners, the understanding of the cerebral interpretation of pain is expanding. Remember as well that emotional and psychological factors are crucial in pain perception, and are the starting points for any individual's perception of a painful circumstance. PMID- 15829383 TI - Pain assessment instruments for use in the emergency department. AB - Each patient's pain experience is uniquely his or her own. Standardized pain assessment methodologies and procedures provide a window to this experience and constitute a necessary first step to our understanding of pain, in both clinical and research settings. All too often, emergency department pain assessment is cursory--performed more to satisfy regulatory requirements than to guide our therapies or evaluate our practices. This article provides information on a number of assessment techniques that are appropriate for clinical and research use. Their use should inform our practice and lead to continuous improvements in our management of pain. PMID- 15829384 TI - Changing attitudes about pain and pain control in emergency medicine. AB - Oligoanalgesia continues to be a large problem in the ED. An attitude of suspicion, a culture of ignoring the problem, and an environment that is not conducive to change in practice combine to present formidable obstacles for effective pain management in the emergency setting. Overcoming these obstacles for effective analgesia in the ED is not beyond the capabilities of the individual ED, the emergency physician, or the specialty of emergency medicine. Changing the attitudes of emergency medical providers about pain assessment and management will require attention in several areas of research, education and training. Oligoanalgesia remains a global problem within emergency medicine; however, this awareness is often not felt to be present "in my ED." Individual ownership of the problem may contribute to improvements in pain control. The last 15 years have seen a substantial increase in ED research focused on pain and pain management. Continued research efforts and focused clinical application of these efforts are still required. A better understanding of patient needs and expectations for pain relief, as well as continued efforts at patient education regarding pain, will also improve our treatment of pain in the ED. Recognition by providers of the ethnic, cultural, and gender differences in the expression, reporting, and expectations for treatment of pain should also continue to be a priority in changing attitudes toward pain and pain control. These goals must be realistic within the chaotic and unpredictable environment that defines emergency medicine. Practical and time-sensitive approaches to pain and pain management will continue to bea challenge to enact and enforce in our EDs. The stigma of opioids, in combination with the transient nature of the emergency physician/patient relationship, may be the largest hurdles to overcome for effective pain management not only in the ED, but also following ED discharge. Improvement in provider education of the realities, myths, and misunderstandings of opioid management may provide insight into this problem. The consequences of oligoanalgesia in the ED are not insignificant. To improve our treatment of pain in the ED, a fundamental change in attitude toward pain and the control of pain is required. This is unlikely to occur until pain is adequately addressed and treated appropriately as a true emergency. PMID- 15829385 TI - Management of chronic pain in the acute care setting. AB - Chronic nonmalignant pain requires evaluation and treatment different from acute pain. The pathophysiology is different, and there is commonly some degree of psychosocial dysfunction. Opioids tend to be much less effective as analgesics for chronic pain, and may increase the sensitivity to pain when given long-term. Because they are self-reinforcing, opioids may be sought and be reported to improve chronic pain, even when they may make the condition worse over time. There are many effective alternatives to opioids for the treatment of chronic pain, but their use is complicated and may require considerable time and effort to determine which ones work. Patients, particularly those who have already been prescribed opioids, may resist these alternatives. An extensive physical and psychosocial evaluation is required in the management of chronic pain, which is difficult if not impossible to achieve in the emergency or urgent care settings. Consequently, emergency and urgent care physicians should work closely with the patient's pain management specialist or personal physician. Systems should be set up in advance to identify those patients whose frequent use of acute care services for obtaining opioids may be compromising their long-term management, putting themselves at risk for psychological and tolerance-induced adverse effects of frequent opioid use. Opioids may be used in carefully selected patients in consultation with their pain management specialist or personal physician, but care must be exercised not to initiate or exacerbate psychological or tolerance-related complications of chronic pain. PMID- 15829386 TI - The psychology of pain. AB - Emotional and evaluative issues are very important in the evaluation and treatment of pain. Treating the physical pain alone can leave these issues unresolved, and possibly exacerbate them through reinforcement. Understanding the impact of fear, expectations, and attention can help physicians deal more effectively with acute pain. Psychological issues are particularly prominent in chronic pain. Though acute care physicians may not be treating these psychological conditions, they can help by referring patients to the appropriate psychological or multidisciplinary setting. PMID- 15829387 TI - The drug-seeking patient in the emergency room. AB - Drug-seeking patients include recreational drug abusers, addicts whose dependence occurred through abuse or the injudicious prescription of narcotics, and pseudoaddicts who have chronic pain that has not been appropriately managed. Opioids produce euphoria in some patients, providing the motivation for abuse, which can be detrimental even with occasional use. Even in the absence of overt euphoria, opioids are highly self-reinforcing and can be problematic in a large number of patients, requiring that acute care physicians exercise caution in whom they are administered. Habitual patient files, narcotic contracts, pain management letters, and patient tracking and management programs can be used for the benefit of both drug seeking-patients and chronic pain patients. For many patients, drug-seekers and chronic pain patients alike, withholding opioids may be an important part of their long-term management. For others, long-acting opioids such as long-acting morphine or methadone are a reasonable option. PMID- 15829388 TI - Painful dilemmas: an evidence-based look at challenging clinical scenarios. AB - Difficult clinical decisions are a part of every emergency practitioner's life. Dealing with difficult patients and recalcitrant consultants is seldom enjoyable, but can be made more palatable through the use of some of the clinical strategies contained in this article. Knowledge of the current best evidence and a willingness to discard outdated practice ideas will help ensure that emergency practitioners continue to provide state-of-the-art medical care. Expressions of care, concern, and respect for patients' problems, and development of a therapeutic alliance with these patients will maximize patient, and ultimately physician, satisfaction. PMID- 15829389 TI - Pediatric pain management in the emergency department. AB - Over the past 25 years, pediatric emergency medicine research and literature have progressively augmented our knowledge of safe and effective pediatric pain management strategies. Yet there is still much more we need to do to understand the painful experiences of children, and to develop optimal safe ways of addressing their needs within the context of a busy pediatric emergency department (ED). In this article, the authors review the history of ED pediatric pain management and sedation, discuss special considerations in pediatric pain assessment and management, review various pharmacologic and nonpharmacologic methods of alleviating pain and anxiety, and present ideas to improve the culture of the pediatric ED, so that it can achieve the goal of becoming pain-free. PMID- 15829390 TI - Pain management in the prehospital environment. AB - Pain measurement and relief is complex and should be a priority for prehospital providers and supervisors. The literature continues to prove that we are poor pain relievers, despite the high prevalence of pain in the out-of-hospital patient population. Lack of education and research, along with agent availability, controlled substance regulation, and many myths given credence by health care providers, hinder our ability to achieve adequate pain assessment and treatment in the prehospital setting. Protocols must be established to help guide providers through proper acknowledgment, measurement, and treatment for prehospital pain. Nonpharmacologic therapies must also be taught and reinforced as important adjuncts to pain management. Finally, formation of quality improvement pain programs that evaluate patient outcomes and provider practice patterns will help EMS systems understand the pain management process and outline areas for improvement. Only through emphasis on pain education, research, protocol and program monitoring development will the quality of pain assessment and management in the prehospital setting improve. PMID- 15829391 TI - Basic pharmacology and advances in emergency medicine. AB - To provide optimal pain control for their patients, emergency physicians should have an in-depth understanding of analgesic drugs and how to use and combine them effectively. The purpose of this article is to describe the pharmacology and use of analgesic agents that are most useful in the management of acute pain in the emergency department. PMID- 15829392 TI - The future of pain management in emergency medicine. AB - How have we as a profession, whose number-one goal is to decrease human suffering, made pain control such a poorly discussed issue in training? From day 1 of medical school, pain and suffering need to be discussed. No clinical area should be taught without discussion of this most common and most important symptom. Although we have shown that up to 70% of our patients have pain as a part of their presenting problem, hospitalized patients also have high rates of pain, often unrecognized. Barriers need to be identified and discussed. Alternatives to medications should be as much a part of our armamentarium as caring and compassion. The future of pain control depends on this paradigm shift. PMID- 15829393 TI - Local anesthetics and peripheral nerve blocks in the emergency department. AB - The emergency physician has a variety of options for providing effective pain relief. A solid understanding of the local anesthetic agents and regional anesthetic techniques is an essential component of every emergency physician's analgesia armamentarium. PMID- 15829394 TI - Procedural sedation and analgesia: a review and new concepts. AB - Procedural sedation and analgesia has become a commonplace procedure in the ED, certainly falling under the domain of the EP. Every EP should approach PSA as a complex procedure requiring high-level skills and knowledge. Initially, understand that PSA represents a spectrum of goals, from anxiolysis and pain relief to deep sedation. Assess the needs of the patient and the concomitant procedure and set goals accordingly. There is a pharmacopia of drugs that provide sedation and analgesia. Become familiar with their pharmacology, advantages and disadvantages, and indications. This will allow for appropriate usage and achievement of sedation goals. Several drugs that are commonly used for general anesthesia are proving themselves to be safe and efficacious for PSA. Both etomidate and propofol have emerged as useful drugs for PSA. Continued research and practice with these agents will add to our understanding and help define their use for PSA. Performing PSA as a procedure itself requires preparedness, diligent monitoring, and risk awareness. Knowing the patient's comorbid state and choosing agents that will not exacerbate their baseline status minimize risk. Following fasting guidelines is appropriate in certain clinical situations, and is prudent when time permits. However, these guidelines are a benchmark for minimizing risk and are not supported by evidence-based medicine. It is important to be cognizant of the guidelines but also to identify the emergency scenario where action must be taken despite the fasting guidelines. Controlling sedation depth also minimizes the risk of aspiration and other complications. The ETCO2 monitor and Bispectral Index may prove to be useful adjuncts for monitoring sedation depth. However, there is nothing yet that measures sedation depth quantitatively that can replace the qualitative assessment of the EP. More and more PSA is falling under the domain of the EP. It is important for the EP to be involved in hospital policy and guidelines associated with this procedure, and to remain aware of new research in this field. EPs can thereby contribute to quality assurance throughout the medical community by setting a standard in the practice of PSA, as they are not the only practitioners using this procedure. With continued practice and research, expertise in this field will grow measurably. PMID- 15829395 TI - Establishing an emergency department pain management system. AB - Any ED system for the management of pain in the ED should consider the following: assessment of pain including mandatory use of some assessment tool, a guideline for treatment of pain, communication with other members of the health care team, assessment tools, program monitoring, and a continuous quality assurance program. The treatment guideline should consider acute versus chronic pain, potential medication tolerance, concurrent anxiety and psychiatric issues, special populations, and disease-specific conditions. PMID- 15829396 TI - Complementary and alternative pain therapy in the emergency department. AB - One primary reason patients go to emergency departments is for pain relief. Understanding the physiologic dynamics of pain, pharmacologic methods for treatment of pain, as well CAM therapies used in treatment of pain is important to all providers in emergency care. Asking patients about self-care and treatments used outside of the emergency department is an important part of the patient history. Complementary and alternative therapies are very popular for painful conditions despite the lack of strong research supporting some of their use. Even though evidenced-based studies that are double blinded and show a high degree of interrater observer reliability do not exist, patients will likely continue to seek out CAM therapies as a means of self-treatment and a way to maintain additional life control. Regardless of absolute validity of a therapy for some patients, it is the bottom line: "it seems to help my pain." Pain management distills down to a very simple endpoint, patient relief, and comfort. Sham or science, if the patient feels better, feels comforted, feels less stressed, and more functional in life and their practices pose no health risk, then supporting their CAM therapy creates a true wholistic partnership in their health care.CAM should be relatively inexpensive and extremely safe. Such is not always the case, as some patients have discovered with the use of botanicals. It becomes an imperative that all providers be aware of CAM therapies and informed about potential interactions and side effects when helping patients manage pain and explore adding CAM strategies for pain relief. The use of regulated breathing, meditation, guided imagery, or a massage for a pain sufferer are simple but potentially beneficial inexpensive aids to care that can be easily employed in the emergency department. Some CAM therapies covered here, while not easily practiced in the emergency department, exist as possibilities for exploration of patients after they leave, and may offer an improved sense of well being and empowerment in the face of suffering and despair. The foundations of good nutrition, exercise, stress reduction, and reengagement in life can contribute much to restoring the quality of life to a pain patient. Adding nondrug therapies of physical therapy, cognitive-behavioral therapy, TENS, hypnosis, biofeedback, psychoanalysis, and others can complete the conventional picture. Adding in simple mind/body therapies, touch therapies, acupuncture, or others may be appropriate in select cases, and depending on the circumstances, may effect and enhance a conventional pain management program. Armed with an understanding of pain dynamics and treatments, practitioners can better meet patient needs, avoid serious side effects, and improve care when addressing pain management in the emergency department. PMID- 15829397 TI - Procedural sedation and analgesia in the emergency department: what are the risks? AB - The practitioner of emergency medicine is routinely faced with patients in need of emergent procedures and pain control and sedation. Our challenge is to make our patients' experiences as painless and as safe as possible, while maximizing our ability to perform the procedure at hand; this is not always an easy task given the propensity of each human body to react differently to interventions and stimuli. We can best meet this challenge by understanding how our patients and pharmaceutical agents intermingle in the risk-benefit equation we formulate before starting our "experiment." Coupling this information with fundamentally sound patient care and monitoring will minimize bad experiences with PSA for both the patient and practitioner. PMID- 15829398 TI - Legal issues in pain management: striking the balance. AB - The momentum over the past several years resulting in positive change to state pain policy is encouraging, especially as the medical community meets greater expectations from patients for appropriate pain management. As the trend for increased medical use of opioid medications continue it is important that medical professionals continue to work with government officials to ensure efforts to curb drug abuse do not impede patient access to pain management. Essential to maintaining that balance is an expansion of scholarly work supporting the medical use of opioids and promotion of research through educational activities. In 2004, the Mayday Project at ASLME will host a national conference, with an emphasis on pain management in the emergency department. The previous noted success of Mayday Project activities should give all of us hope that through scholarly activity further guidance will be provided in finding that critical balance between effective pain management and drug diversion. PMID- 15829399 TI - Pain management in the wilderness and operational setting. AB - The wilderness and operational setting places unique constraints on one's ability to treat pain. In this article we will discuss methods for treating pain both in the wilderness and operational setting. By operational we mean the austere deployed military setting, to include both noncombat and combat operations. The authors combined experience with wartime trauma pain management consists of experience in Operation "Just Cause" (Panama Invasion), Operation "Desert Storm" (Persian Gulf War), Operation "Uphold Democracy" (Haiti liberation), Operation "Enduring Freedom" (Afghanistan conflict), and Operation "Iraqi Freedom" (Iraq conflict). PMID- 15829400 TI - High incidence of spontaneous autoimmune thyroiditis in immunocompetent self reactive human T cell receptor transgenic mice. AB - Autoantigen-specific TCR transgenic mice allow us to assess the role of T cells in autoimmunity. We have recently generated humanized TAZ10 transgenic mice expressing the human TCR specific for the immunodominant epitope of thyroid peroxidase (TPO). We have shown that these transgenic mice do not undergo tolerance in vivo and that on Rag deficient background they are susceptible to spontaneous autoimmune thyroiditis. Here we show that, in contrast to other transgenic models of autoimmunity, almost all TCR(+)Rag1+ (T+R+) T cells are activated in vivo leading to the development of spontaneous autoimmune thyroiditis. In these mice, disease is also accompanied by a significant reduction of CD4+CD25hi regulatory T cells. These data indicate that the pathogenic activity of the self-reactive TCR can circumvent the regulatory function operated by the non-transgenic T cells that are normally present in T+R+ mice, leading to autoimmunity. PMID- 15829401 TI - Infection-induced apoptosis deletes bystander CD4+ T cells: a mechanism for suppression of autoimmunity during BCG infection. AB - Infection with Mycobacterium bovis Bacille Calmette Guerin (BCG) induces high levels of apoptosis among activated CD4+ T cells. We have investigated the specificity of this pro-apoptotic response and its influence on CD4+ T cell mediated autoimmunity. Apoptosis induced by BCG-infection is unrelated to antigenic specificity, as demonstrated by the increased apoptosis of activated TCR transgenic CD4+ T cells of unrelated specificity. Moreover, infection-induced apoptosis promoted the deletion of CD4+ T cells activated either by peptide or anti-CD3/anti-CD28 stimulation. Infection-induced apoptosis required IFN-gamma production by the infected host, and expression of the IFN-gamma receptor on donor CD4+ T cells. We used an adoptive transfer model of experimental autoimmune encephalomyelitis (EAE) to assess the influence of infection-induced apoptosis on a CD4+ T cell-mediated response. A significantly higher level of apoptosis was seen among sorted encephalitogenic CD4+ T cells transferred to BCG-infected versus uninfected hosts. BCG-infected mice displayed a milder course of clinical disease than their uninfected counterparts and a decreased recovery of donor cells from the CNS. The data suggest that mycobacterial infection attenuates the severity of EAE, at least in part, by promoting the apoptotic elimination of autoreactive CD4+ T cells. PMID- 15829402 TI - How an autoimmune reaction triggered by molecular mimicry between streptococcal M protein and cardiac tissue proteins leads to heart lesions in rheumatic heart disease. AB - Molecular mimicry between microbial antigens and host tissue is suggested as a mechanism for post-infectious autoimmune disease. In the present work we describe the autoimmune reactions of two severe rheumatic heart disease (RHD) patients, through an analysis of heart-infiltrating T-cell repertoire, antigen recognition, and cytokine production induced by specific antigens. T-cell clones derived from oligoclonally expanded T cells in the heart cross-recognized M5 peptides, heart tissue-derived proteins, and myosin peptides. We show, using binding affinity assays, that an immunodominant streptococcal peptide (M5(81-96)) is capable of binding to the HLA-DR53 molecule. The same peptide was recognized by an infiltrating T-cell clone from a patient carrying HLA-DR15, DR7, and DR53 molecules. This suggests that this peptide is probably presented to T cells in the context of the HLA-DR53 molecule. Cross-reactive heart-infiltrating T cells activated by the M5 protein and its peptides and by heart tissue-derived proteins produced predominantly inflammatory cytokines. Interleukin (IL)-4 was produced in small amounts by mitral valve intralesional T-cell lines and clones. Altogether, these results suggest that mimicry between streptococcal antigens and heart tissue proteins, combined with high inflammatory cytokine and low IL-4 production, leads to the development of autoimmune reactions and cardiac tissue damage in RHD patients. PMID- 15829403 TI - T-cell molecular mimicry in Chagas disease: identification and partial structural analysis of multiple cross-reactive epitopes between Trypanosoma cruzi B13 and cardiac myosin heavy chain. AB - Chagas disease cardiomyopathy (CCC) is one of the few examples of post-infectious autoimmunity, where infectious episodes with an established pathogen, the protozoan parasite Trypanosoma cruzi, clearly triggers molecular mimicry-related target organ immune damage. CD4+ T-cell clones infiltrating hearts from CCC patients cross-reactively recognize human cardiac myosin, the major heart protein, and the immunodominant B13 protein from T. cruzi. Moreover, in vitro priming with B13 leads to the recovery of cardiac myosin cross-reactive T-cell clones. In order to identify cross-reactive epitopes between B13 protein and human cardiac myosin, we used B13 peptide S15.4, preferentially recognized by CCC patients, to establish a T-cell clone from an HLA-DQ7 individual. The B13 S15.4 peptide-specific CD4+ T-cell clone 3E5 was tested in proliferation assays against 15 Lys/His-substituted S15.4-derived peptides for TCR/HLA contact analysis. Together with previous HLA-binding data and molecular modeling of the HLA-DQ7 peptide S15.4 complex, Lys/His scanning analysis showed eight TCR/HLA contact positions. Clone 3E5 was also tested against 45 15-mer peptides from human beta cardiac myosin heavy chain bearing the central HLA-DQ7 binding motif. Clone 3E5 recognized 13 peptides from cardiac myosin. The alignment of cross-reactive peptides in cardiac myosin showed very limited sharing of residues or side chains with similar chemical/structural features at aligned positions, indicative of a very degenerate TCR recognition pattern. The existence of degenerate intramolecular recognition, with multiple low-homology, cross-reactive epitopes in a single autoantigenic protein may have implications in increasing the magnitude of the autoimmune response in CCC and other autoimmune diseases. PMID- 15829404 TI - Th1 and Th2 lymphocyte migration across the human BBB is specifically regulated by interferon beta and copolymer-1. AB - Lymphocyte migration into the central nervous system is a central event in lesion formation in MS. Both interferon beta (IFNbeta) and copolymer-1 (Cop-1) reduce the overall lymphocyte entry into the brain through the blood-brain barrier (BBB) as judged by MRI based studies. In this study, we used a modified Boyden chamber assay in which human brain microvascular endothelial cell (HBEC) monolayers are grown on a fibronectin coated transwell membrane to evaluate in vitro migration of allo-antigen Th1 and Th2 lymphocytes across brain endothelium. We confirmed previous observations showing that migration rates of Th2 lymphocytes across HBECs were higher than migration rates of Th1 cells. When HBECs were pre-treated with IFNbeta (100 U/ml) 30 min prior to migration, the migration rate of Th1 was significantly decreased (45% reduction) while the migration of Th2 remained unchanged. Addition of Cop-1 (30 microg/ml) to HBEC monolayers 30 min prior to migration significantly increased the migration rate of Th2 cells and did not affect the migration of Th1 cells. We did not observe any changes in (1) the expression of adhesion molecules on the surface of HBECs and (2) the pattern of chemokine production by HBECs after IFNbeta or Cop-1 treatment. The changes in cellular migration rates were not paralleled with changes in diffusion of large molecular weight tracers across brain ECs. Our data support the notion that immuno-modulators used for the treatment of MS selectively and differentially regulate the migration of T helper lymphocyte subsets and that Cop-1 promotes trans-endothelial migration of Th2 cells across the BBB. PMID- 15829405 TI - G-CSF treatment prevents cyclophosphamide acceleration of autoimmune diabetes in the NOD mouse. AB - Cyclophosphamide (CY) accelerates autoimmune diabetes in the NOD mouse at different levels, including critical targeting of a regulatory T cell subset, exacerbation of pro-Th1 IFN-gamma production and promotion of inflammation in pancreatic islets. Here we evaluated the ability of G-CSF to antagonize the acceleration of the disease induced by CY. Human recombinant G-CSF, administered daily at 200 microg/kg by s.c. injection, protected NOD mice from CY-accelerated onset of glycosuria and insulitis. G-CSF accelerated the recovery of the T cell compartment after the depletion of the lymphoid compartment triggered by CY injection. It selectively prevented the loss of the immunoregulatory T cells expressing the CD4(+)CD25+ phenotype that also stained CD62L+ in peripancreatic lymph nodes and promoted their expansion in the spleen. In addition to this, it abrogated the robust cytokine--particularly IFN-gamma- and chemokine burst triggered in immune cells by CY. G-CSF promoted only slight changes in the inflammatory effects of CY at the target tissue site, assessed by chemokine induction within the pancreas. Thus the immunoregulatory properties of G-CSF were critical in the early control of the accelerating effects of CY on autoimmune diabetes in the NOD mouse. PMID- 15829406 TI - Monoclonal ribosomal P autoantibody inhibits the expression and release of IL-12, TNF-alpha and iNOS in activated RAW macrophage cell line. AB - Monoclonal ribosomal P protein antibody (anti-P mAb) may bind to the cell surface, penetrate into cells, and induce apoptosis of Jurkat T cells. Recently, modulation of cytokines has been considered to be important in the pathogenesis of systemic lupus erythematous (SLE). In this study, effects of anti-P mAbs (9B6) on gene expression of cytokines, apoptosis, and reactive oxygen species in murine macrophage RAW 264.7 were analyzed by RT-PCR and ELISA and those on IL-12 promoter activity was determined in an IL-12p40 promoter-reporter gene transfected cell line RAW (IL-12p40-SEAP). After treating LPS-activated RAW 264.7 with 9B6 for 6 or 24 h, the levels of mRNA and protein expression of IL-12, TNF alpha, and iNOS were significantly inhibited by 25%, 16%, and 13%, respectively. The IL-12 promoter activity of RAW (IL-12p40-SEAP) was also inhibited by 13-22%. However, inhibitory effects were not observed in cells pre-treated with IgG1 for 1 h. The productions of IL-10 in LPS-activated RAW 264.7 and human macrophages were potentiated by 9B6 up to 65% and 51%, respectively. Since anti-P Abs inhibit productions of IL-12 and TNF-alpha and enhance IL-10 production in macrophages, these autoantibodies may augment Th2 responses and amplify lupus manifestations by causing immunological polarity and lymphocyte dysfunction. PMID- 15829407 TI - Nuclear localizing anti-DNA antibodies enter cells via caveoli and modulate expression of caveolin and p53. AB - After administration to normal mice, a subset of monoclonal (m) anti-DNA antibodies (Ab) derived from MRL-lpr/lpr mice was identified that enter cells, in vivo. In the kidneys, this was associated with glomerular hypercellularity and proteinuria. In cultured cells, the same mAb bound to myosin 1 on the cell surface, prior to internalization, nuclear localization and inhibition of apoptosis. The present study focuses on the mechanisms underlying the observed functional effects. Subcellular localization studies revealed that following internalization, a prototypic, nuclear localizing, m antibody (Ab; termed H7) co localized with myosin 1, shortly after internalization, within caveolae, near the cell membrane. Cell fractionation studies confirmed the presence of both H7 and myosin within the caveolar fraction. Since variations in caveolin protein expression have been associated with apoptotic events in cancer cells, through p53 dependent and independent pathways, modulation of caveolin by intracellular H7 was evaluated. Cellular entry of the anti-DNA Ab resulted in an increase in caveolin protein expression. Furthermore, after exposure of cells to dexamethasone to induce apoptosis, the usual increase in p53 was inhibited in the presence of intracellular H7. Taken together, the results suggest that upregulation of caveolin and inhibition of p53 induction are involved in H7 induced, inhibition of apoptosis. Furthermore, they suggest that this inhibition contributes to the glomerular hypercellularity observed in normal mice with intranuclear H7. The results also raise the possibility that inhibition of apoptotic pathways during inflammation or/and autoimmunity could influence subsequent disease events. The novel mechanism of cellular perturbation is indirect and dependent on apoptotic stimuli, and it may account for the presence of intranuclear antibodies in inflammatory and normal tissues of individuals with lupus. PMID- 15829408 TI - Absence of avidity maturation of autoantibodies to the protein tyrosine phosphatase-like IA-2 molecule and glutamic acid decarboxylase (GAD65) during progression to type 1 diabetes. AB - Immunoglobulin G avidity assays are used to distinguish between the acute and chronic phase of several infectious diseases, and there is evidence of autoantibody affinity maturation also in autoimmune diseases. To assess whether the analysis of the avidity of autoantibodies against the protein tyrosine phosphatase-like IA-2 molecule and glutamic acid decarboxylase (GAD65) could improve the accuracy of risk assessment of progression to clinical type 1 diabetes, we established methods for the determination of the autoantibody avidity based on our previously developed time-resolved fluorometric IA-2 and GAD65 autoantibody (IA-2A and GADA) assays. The avidity indices of sequential plasma samples from six IA-2A-positive and seven GADA-positive prediabetic children were analysed applying elution with urea and diethylamine (DEA). For comparison, corresponding avidity indices of control children, who have remained non-diabetic for at least 3 years after seroconversion to IA-2A and GADA positivity, were analysed. For most of the children, only a slight fluctuation in the avidity index values was observed over time, although the titres for IA-2A and GADA varied substantially in some cases. The avidity indices of the prediabetic children remained within the same range as those of the control group throughout the follow-up. Our results indicate that the analysis of the avidity index levels of IA-2A and GADA does not improve the accuracy of the prediction of type 1 diabetes based on autoantibody detection. PMID- 15829409 TI - Analysis of autoantibody repertoires in small- and medium-sized vessels vasculitides. Evidence for specific perturbations in polyarteritis nodosa, microscopic polyangiitis, Churg-Strauss syndrome and Wegener's granulomatosis. AB - In order to identify new antibody reactivities, we have used a quantitative immunoblotting technique on extracts of normal human tissues to analyze the repertoires of serum IgM, serum IgG and purified IgG autoantibodies of patients with systemic vasculitides. Patients fulfilled the American College of Rheumatology and Chapel Hill criteria for the diagnosis of polyarteritis nodosa (PAN) (n=8), PAN related to hepatitis B virus (HBV) infection (n=5), Wegener's granulomatosis (WG) (n=6), microscopic polyangiitis (MPA) (n=18) or Churg-Strauss syndrome (CSS) (n=8). Sera from patients with chronic HBV infection without PAN (n=5) and age- and gender-matched healthy individuals (n=45) were used as controls. In the lung extract, IgM from 12/18 MPA patients reacted with high intensity with a 50 kDa band and serum IgG from 3/8 CSS patients bound to a 70 kDa protein band. In the artery extract, serum IgG from 6/18 MPA patients bound to an 85 kDa antigen, whereas purified IgG from all WG patients tested bound to a 28 kDa protein band and IgM from CSS patients bound to 2 main antigens of 38 and 60 kDa. These results provide evidence for the specificity of autoantibody repertoires from patients with PAN, WG, CSS and MPA. PMID- 15829410 TI - Immunomodulatory activity of analog of muramyl dipeptide and their use as adjunct to chemotherapy of Leishmania donovani in hamster. AB - In search of a potent immunomodulator to be used as an immunoprophylactic agent and as adjunct to chemotherapy against Leishmania infection, two analogs of muramyl dipeptide, viz. N.Ac-norMur-MeVal-D-isoGln (86/448) and N.AcMur-Acc-D isoGln (89/729) were evaluated for desired activity. Effect of these peptides on cell mediated and humoral immunity was studied by immunizing the peptide treated mouse with sheep red blood cells (SRBC) and determining HA-titer, plaque forming cells assay and delayed type of hypersensitivity (DTH) response after 4-5 days. Both the peptides stimulated cell mediated immunity (CMI), humoral response as well as macrophage function in terms of super oxide anion (O2-) and nitric oxide (NO) generation. Mitogen induced lymphocyte proliferation and production of IL-2 and INF-gamma increased while that of IL-4 and IL-10 decreased by both the peptides showing a typical Th1 type response. After establishing the immunostimulatory activity, these peptides were evaluated for immunoprophylactic efficacy as well as for use as adjunct to chemotherapy with stibanate (SSG) against Leishmania donovani infection in golden hamster. These peptides were found quite effective in both the modes. In adjunct use the treatment may require lower dose of SSG and thereby reduce the chances of drug toxicity. PMID- 15829411 TI - Systemic treatment with n-6 polyunsaturated fatty acids attenuates EL4 thymoma growth and metastasis through enhancing specific and non-specific anti-tumor cytolytic activities and production of TH1 cytokines. AB - Recently, there has been a great interest in the effects of different types of n 6 polyunsaturated acids (n-6 PUFAs) upon the immune system and cancer development. However, the effects of n-6 PUFAs are still controversial and as yet undefined. The present study aimed to investigate the anti-tumor effects of n-6 PUFAs against EL4 thymoma and the associated immune mechanisms. To this, sesame oil, a vegetable oil enriched with n-6 PUFAs, or free linoleic acid (LA) were administered intraperitoneally into C57BL/6 mice before and after challenge with EL4 lymphoma cells. Treatment with either sesame oil or LA attenuated the growth and metastasis of EL4 lymphoma. The anti-tumor effect of LA was superior to that of sesame oil, and associated with an increase in the survival rate of the tumor bearing mice. In addition, both sesame oil and LA showed dose-dependent anti lymphoma growth in vitro. Treatment with LA generated significant increases in the anti-lymphoma cytolytic and cytostatic activities of T cells and macrophages, respectively, and enhanced production of IL-2 and IFN-gamma while decreased production of IL-4, IL-6 and IL-10. In summation, the results suggest that n-6 PUFAs, represented by LA, can attenuate EL4 lymphoma growth and metastasis through enhancing the specific and non-specific anti-tumor cytolytic activities and production of TH1 cytokines. These findings might be of great importance for a proper design of systemic nourishment with PUFAs emulsions for cancer patients. PMID- 15829412 TI - Low molecular weight fraction of shark cartilage can modulate immune responses and abolish angiogenesis. AB - Shark cartilage has proven to have inhibitory effects on angiogenesis. In this research, we studied the effects of shark cartilage on the immune system. Firstly, we isolated and purified a shark cartilage protein fraction with the most immunostimulatory effects. Our fraction was composed of two proteins with molecular weights of about 14 and 15 kDa. This fraction highly augments delayed type hypersensitivity response against sRBC in mice, and decreases the cytotoxic activity of Natural Killer cells. Furthermore, intraperitoneal injection of this fraction to tumor-bearing mice could increase T-cell infiltration into the tumor, and decrease the tumor lesion size. Also, this fraction has strong inhibitory effect on HBMEC proliferation and migration in fibrin matrix. According to these results, we suppose that this fraction is a good candidate for further studies in cancer therapy. PMID- 15829413 TI - Transduction of the gene coding for a human G-protein coupled receptor FPRL1 in mouse tumor cells increases host anti-tumor immunity. AB - Low antigenicity or development of tolerance is believed to be a major contributor to the escape of malignant tumors from immune surveillance of the host. However, anti-tumor responses can be elicited by concomitant immunization of poorly antigenic tumor cells with homologous xenogeneic proteins as 'altered self' proteins. In our study, anti-tumor, but not anti-xenogeneic antigen, immune responses were generated after transduction of the gene coding for a G-protein coupled human formyl peptide receptor like-1 (FPRL1) into a mouse C26 colon cancer cell line. C26 cells transfected with FPRL1 gene exhibited markedly reduced tumorigenicity in syngeneic mice, in association with the appearance of high levels of antibody activity reacting with both FPRL1 containing and wild type C26 cells. The anti-tumor responses required the participation of CD4+ T lymphocytes, since no tumor rejection was observed in nude mice or in syngeneic mice depleted of CD4+ T cells. Furthermore, mice primed with FPRL1 transfected C26 cells were resistant to subsequent challenge by wild type C26 cells. These results indicate that the presence of human FPRL1 is capable of triggering specific anti-tumor host immune responses against poorly antigenic mouse tumor cells. PMID- 15829414 TI - Oligodeoxynucleotides containing synthetic immunostimulatory motifs augment potent Th1 immune responses to HBsAg in mice. AB - Toll-like receptor 9 (TLR9) modulators have potent Th1-adjuvant activity. We recently reported the development of immunomodulatory oligonucleotides (IMOs) containing novel structures (immunomers) and synthetic immunostimulatory CpR (R=2'-deoxy-7-deazguanosine) or R'pG (R'=1-(2'-deoxy-beta-D-ribofuranosyl)-2-oxo 7-deaza-8-methyl-purine) motifs. IMOs activate TLR9 pathways, resulting in cytokine secretion profiles different from those induced by CpG DNA. In the present study we evaluated the adjuvant activity of IMOs containing CpG, CpR, or R'pG motifs in combination with hepatitis B surface antigen (HBsAg) in a mouse model. Mice immunized with HBsAg plus IMO produced higher levels of IgG2a and lower levels of IgG1 than did mice immunized with HBsAg alone or with alum. High IgG2a responses were found at week 4 and remained high until 14 weeks after immunization. Adoptive transfer of splenocytes from HBsAg/IMO-immunized mice to naive mice resulted in strong IgG2a production in response to antigen boost. Splenocytes of mice immunized with HBsAg/IMO produced high levels of IFN-gamma, but not Th2 cytokines IL-4 and IL-5, in antigen-recall experiments in vitro. The use of IMOs as adjuvants to HBsAg resulted in the production of strong anti-HBsAg antibodies at antigen doses as low as 0.2 microg. These data demonstrate that IMOs enhance the immunogenicity of HBsAg through potent Th1 immune responses, which may allow lower doses of antigen in vaccination. PMID- 15829415 TI - Development and validation of a novel IL-10 deficient cell transfer model for colitis. AB - A number of rodent models for inflammatory bowel disease (IBD) have been developed, but most cannot be used to develop and validate new therapies for IBD. From the models developed, the IL-10 deficient mouse model is the one that results in a disease similar to human IBD; however, in this model, colitis occurs with variable incidence taking 3-4 months to develop. These are serious problems with the model when evaluating a new therapy because of the large-scale experiments required and the difficulty in performing an accurate pharmacological analysis. In this study, the IL-10 deficient mouse model was modified by transferring whole spleen and mesenteric lymph node cells from IL-10 deficient mice to CB-17 SCID mice. In this IL-10 deficient cell transfer model, chronic intestinal inflammation developed in all recipients within 2-3 weeks, which was far earlier than in donor IL-10 deficient mice. The pathological phenotypes were similar to those of IL-10 deficient mice and CD45RBhi T cell-transfer models. In addition, we assessed several agents for inflammatory bowel disease to validate the general utility of this cell transfer model. It is worth noting that TNFR-Ig or prednisolone, which is effective for treatment of patients with severe fulminant Crohn's disease, markedly attenuated pathological clinical indices in this colitis model, whereas the immunosuppressive agents, azathioprine, tacrolimus, and cyclosporine A produced no significant effect. These results suggest that the IL-10 deficient cell transfer model is a good experimental model to use for developing new and effective therapies for active IBD. PMID- 15829416 TI - The screening and isolation of an effective anti-endotoxin monomer from Radix Paeoniae Rubra using affinity biosensor technology. AB - Lipopolysaccharide (LPS) is a known trigger in the pathogenesis of sepsis, lipid A being the toxic component. One of several adjuvant therapeutic approaches for severe sepsis is currently focusing on the neutralization of LPS. In order to obtain the components from traditional Chinese herbs that can neutralize the endotoxin, aqueous extractions were tested using affinity biosensor technology. From amongst 42 herbs, eight were found to possess lipid A-binding abilities. Radix Paeoniae Rubras had the highest lipid A-binding ability; therefore an aqueous extraction from this plant was investigated further. After preparation using standard methods, including silica gel chromatography and HPLC, we obtained 1, 2, 3, 4, 6-beta-d-pentagalloylglucose (PGG), with lipid A-binding ability. It was found that in vitro, PGG directly bound to lipid A, with a Kd of 32 microM, and that it neutralized the endotoxin both in the Limulus Amebocyte Lysate (LAL) assay and in a TNF-alpha release experiment, in a dose-dependent manner. In in vivo experiments, PGG was found to protect mice from a lethal challenge by LPS, and significantly decreased the plasma endotoxin level both in endotoxemic mice and rats, the reduction of the endotoxin level in rats being tightly associated with the TNF-alpha level. In conclusion, we demonstrate the effectiveness of affinity biosensor technology in discovering useful agents amongst traditional Chinese herbs and using this approach we found a new anti-endotoxin agent. PMID- 15829417 TI - Detection of opioid receptors on murine lymphocytes by indirect immunofluorescence: mature normal and tumor bearing mice lymphocytes. AB - Opioid peptides modulate immune responses via ligation to classical opioid receptors (mu, delta and kappa), expressed on immune cells, or in an indirect fashion via the central nervous system. The combination of immunofluorescent technique and flow cytometry has proven to be sensitive methods for detection of opioid receptors on leukocytes. In the current study a fluorescein isothiocyanate conjugated naltrexone (FITC-NTX) derivative in the absence or presence of naltrexone, as a competitor, was used to detect opioid receptors on thymocytes and then on splenocytes of normal and tumor bearing Balb/c mice. Tumor bearing mice were made by intraperitoneal injection of fibrosarcoma cell line. In a two weeks interval, tumor grew and then mice splenocytes were harvested. Cells were incubated with FITC-NTX alone (direct fluorescence), or FITC-NTX followed by biotin-conjugated anti-fluorescein IgG and extravidin-R-phycoerythrin (indirect immunofluorescence). Using flow cytometry we found that, with direct fluorescence staining there is only nonspecific cell staining. In contrast, indirect staining of cells demonstrated labeling of opioid receptors. Thymocytes displayed 37.5+/ 7% specific labeling by current staining procedure. However, this specific staining was 17.2+/-4% and 7.5+/-2% in splenocytes of normal and tumor bearing mice, respectively. Taken together, these results showed that, direct fluorescence staining failed to stain opioid receptors expressed on lymphocytes. These receptors can only be detected by a biotin-streptavidin amplification procedure. We also found that the level of opioid receptors on mature lymphocytes is less than that of immature ones and are even lesser in the tumor bearing mice lymphocytes. PMID- 15829418 TI - Effects of mycophenolic acid on endothelial cells. AB - Mycophenolate mofetil (MMF) is a potent immunosuppressant that inhibits the activity of inosine monophosphate dehydrogenase (IMPDH), the rate-limiting enzyme in de novo synthesis of guanosine nucleotides. MMF has been used widely in solid organ transplantation. Increased evidence indicated that MMF exhibited beneficial effects on various types of vasculitis, for reasons that were not fully understood. Endothelial cells play a pivotal role in the pathogenesis of vasculitis. Endothelium may not only be the main target for injury, but also be able to amplify the inflammatory response by adhesion molecule expression, leukocyte adhesion, cytokine production and angiogenesis. In the present study, the effect of mycophenolic acid (MPA), the active metabolite of MMF, on human umbilical vein endothelial cells (HUVECs) was investigated. MPA markedly inhibited tumor necrosis factor-alpha (TNFalpha)-induced intercellular adhesion molecule-1 (ICAM-1) mRNA and surface expression, suppressed TNFalpha-induced neutrophils adhesion to endothelial cells, and reduced TNFalpha-induced interleukin-6 (IL-6) secretion. The inhibitory effects of MPA on ICAM-1 surface expression and IL-6 secretion were not attenuated by addition of guanosine, implying that inhibition of these processes were not due to intracellular guanosine nucleotides depletion. MPA also decreased angiogenesis of endothelial cells in three-dimensional collagen gel culture system, reduced the migration in a wounded monolayer of endothelial cells, and inhibited the proliferation of endothelial cells. In conclusion, MPA exhibited multifarious effects on endothelial cells including inhibition of ICAM-1 expression, neutrophil attachment, IL-6 secretion, and the process of angiogenesis, which might contribute to the efficacy of MMF in the treatment of vasculitis. PMID- 15829419 TI - Protective effects of a Traditional Chinese Medicine, You-Gui-Wan, on steroid induced inhibition of cytokine production in mice. AB - You-Gui-Wan (YGW), a classical yang-tonic prescription of Traditional Chinese Medicine (TCM), is thought to boost the body function against diseases. To determine the effects of YGW on the steroid-induced inhibition of cytokine production in mice, we established a murine model with hydrocoticoid (HC)-induced cytokine suppression. The results showed that oral administration of YGW protected mice against HC-induced inhibition of IFN-gamma, IL-2, IL-4 and IL-10 transcription, and the percentages of CD4+ and CD8+ T cells containing intracellular cytokines including IFN-gamma, IL-4 and IL-10 were significantly increased in murine spleen. The protection of YGW against HC-induced inhibition of cytokine production was further confirmed by the elevated serum level of IFN gamma in YGW-treated mice. The results suggest that YGW improve the immune function even in the serious immunosuppressive condition. PMID- 15829420 TI - Ginkgo terpene component has an anti-inflammatory effect on Candida albicans caused arthritic inflammation. AB - The Ginkgo biloba extract, EGb 761, contains flavonoid glycosides and unique terpene lactones as major active components. In this study, we determined the anti-inflammatory effect of the water-soluble portion (GH415) of the EGb 761 on the inflammation caused by Candida albicans, a major ethiological agent that causes fungal arthritis. For inflammatory induction, an emulsified mixture of C. albicans cell wall and Complete Freund's Adjuvant (CACW/CFA) was injected into BALB/c mice by the hind footpad route once a day for 3 days. Twenty-four hours after the final injection, mice having the swollen footpad were given the GH415 (2 mg/dose) intraperitoneally to the mice once every 3 days for 15 days. The footpad-swelling of these mice was measured during the entire observation period. Results showed that the GH415 treatment reduced the swelling. In the same animal model, this effect was enhanced by treatment with the GH415 entrapped within liposome (Lipo-GH: 200 micro/dose). Further analysis revealed that terpene, not flavone portion, was responsible for such therapeutic anti-inflammatory effect. Treatment with the terpene (7.4 microg/dose) by liposomal delivery method had similar effects as the treatment with indomethacin at 30 microg/dose. Addition of the terpene to lipopolysaccharide-treated macrophages showed suppression of nitric oxide (NO) production. These results suggest that blockage of the NO production from the macrophages that infiltrated to the inflamed site may be a possible mechanism for the therapeutic anti-inflammatory effect. PMID- 15829421 TI - Opposing effect of IFNgamma and IFNalpha on expression of NKG2 receptors: negative regulation of IFNgamma on NK cells. AB - The effector functions of natural killer (NK) cells are regulated by integrated signals across an array of stimulatory and inhibitory receptors interacting with target cell surface ligands. The regulatory effect of interferon-alpha (IFNalpha) and interferon-gamma (IFNgamma) on expression of the family of NKG2 receptors, stimulatory NKG2D receptor and inhibitory NKG2A receptor, and cytolysis of the target tumor cells (MICA+ and HLA-E+) were studied. Results show that IFNgamma and IFNalpha influence NK cell function differently. Interferon-alpha stimulates expression of stimulatory NKG2D receptors and inhibits the expression of inhibitory NKG2A receptors on NK cells. Contrary to the stimulatory effect of IFNalpha, IFNgamma inhibits cytolysis by NK cells of tumor cells expressing MICA or HLA-E cell surface proteins. Blocking NKG2D or NKG2A receptor activity with monoclonal antibodies partly attenuates the inhibitory effect of IFNgamma while promoting the effects of IFNalpha on NK cytolysis. These results show for the first time that IFNgamma negatively regulates NK cells through NKG2 receptors, and that the balance between stimulatory and inhibitory signals through the NKG2 family of receptors may be controlled by two opposing interferons. Modulating the balance between stimulatory and inhibitory signals through cell surface receptors on NK cells may open a new approach to NK cell-based biotherapy for cancer and infectious diseases. PMID- 15829422 TI - Effect of mirtazapine treatment on serotonin transporter in blood peripheral lymphocytes of major depression patients. AB - Lymphocytes from human peripheral blood exhibit a series of markers of neurotransmitters, such as specific receptors and transporters. A reduction of serotonin transporters and an increase of them has been reported after treatment with fluoxetine in depressed patients. The aim of this study was to determine if the administration of an antidepressant with a different mechanism of action, such as mirtazapine, could produce a similar effect. Twenty eight patients (age 41.40+/-2.45) were diagnosed following the criteria for major depression by the Structured Clinical Interview for Disorders of Axis I of the American Psychiatric Association. Severity was measured by Hamilton Scale and by Beck Inventory for Depression, scores of 30.88+/-7.48 and 30.24+/-10.88, respectively, prior to treatment. Samples from control subjects were obtained alternating with patients before and after the administration of the antidepressant: twenty eight and twenty four, respectively (age 38.80+/-2.95). Mirtazapine was given in a dose of 30 mg/day for 6 weeks. Blood lymphocytes were isolated by density gradient from patients and controls before and after treatment. There was a partial response according to clinical evaluation and scores of the Scale and the Inventory. Serotonin transporters were labeled with [3H] paroxetine. Number of sites (B(max)) were 10.86+/-2.60 and 12.58+/-2.71 fmol/10(6) cells for both groups of controls. The depressed patients had a significant reduction of serotonin transporters in their lymphocytes before treatment and an increase after it, with B(max) values of 6.52+/-0.49 and 15.61+/-0.49 fmol/10(6) cells, respectively. There were no significant differences in the affinity for the ligand. Concentrations of serotonin or noradrenaline in lymphocytes were not modified before the treatment, although there was a significant decrease after taking 30 mg/day of the antidepressant for 6 weeks. Mirtazapine, not being a serotonin reuptake inhibitor, did increase the number of transporters in lymphocytes of major depression patients, indicating a complex mechanism, not only directly related to the transporter, but involved in the therapeutic response. PMID- 15829423 TI - Activation of the complement system and leukocyte recruitment by Tityus serrulatus scorpion venom. AB - The scorpion Tityus serrulatus is considered one of the most dangerous species in Brazil. Its venom evokes an inflammatory response, although the exact mechanism of this effect is still unknown. The aim of the present study was to investigate the effect of Tityus serrulatus venom (TsV) on the complement system (CS) and on leukocyte recruitment. Complement consumption by TsV was evaluated using in vitro hemolytic assays, immunoelectrophoresis and two-dimensional immunoelectrophoresis of complement components (factor B and C3). In order to evaluate neutrophil migration induced in normal human serum (NHS) in the presence of TsV, in vitro chemotaxis assays were performed using the Boyden chamber model. In vitro TsV induced a concentration- and time-dependent reduction in hemolytic activity of the classical/lectin and alternative complement pathways, with samples of 43.0 microg and 43.4 microg, respectively, inhibiting 50% of the lytic activity. Alterations in C3 and factor B electrophoretic mobility after incubation of NHS with TsV, were identical to those obtained with zymosan (positive control). Incubation of NHS with TsV induced neutrophil chemotaxis similar to that observed with zymosan-activated serum. Our results show that TsV activates the CS, leading to factor B and C3 cleavage, to reduction of serum lytic activity and generation of complement chemotactic factors. Therefore, CS may play an important role in the inflammatory response observed upon scorpion envenomation. PMID- 15829424 TI - Leflunomide reduces nitric oxide production in patients with active rheumatoid arthritis. AB - OBJECTIVES: Leflunomide is an immunomodulatory agent that was recently approved for the treatment of rheumatoid arthritis (RA). The mechanism of action is not fully understood. Nitric oxide (NO) plays an important role in the pathogenesis of RA. Leflunomide has been shown to cause cell specific inhibition of inducible nitric oxide synthase (iNOS) activation in animal models. We carried out this study to determine if there was alteration in NO production in patients with RA. METHODS: An 8-week open label study was carried out on patients with adult onset active RA. We measured levels of nitrite and citrulline spectrophotometrically as surrogate markers of NO production. Within-patient serum levels of nitrite and citrulline were compared with leflunomide therapy at three points of time (at 0, 4 and 8 weeks of therapy). RESULTS: Thirty-three patients with active RA were enrolled for this study. These patients were a subset of 63 individuals who are studied for clinical efficacy of leflunomide. Three patients were lost to follow up. Median nitrite levels were 817.2 (nmol/ml) at the start of therapy and this declined to 440.9 nmol/ml and 301.1 nmol/ml at 4 and 8 weeks of therapy. Median citrulline levels were 649.3 nmol/ml at the start of the study, which declined to 549.2 nmol/ml and 485.4 nmol/ml at 4 and 8 weeks, respectively. Statistically significant decrease in median values for serum nitrite and citrulline levels was documented after 4 weeks of leflunomide therapy (p<0.01), which was sustained at 8 weeks (p<0.01), although there was no further fall between 4 and 8 weeks (p>0.1). CONCLUSIONS: Leflunomide inhibits nitric oxide production in patients with active RA. Inhibition of NO synthesis may be one of the mechanisms responsible for the immunomodulatory activity of leflunomide. PMID- 15829425 TI - Effect of probiotics on preventing disruption of the intestinal microflora following antibiotic therapy: a double-blind, placebo-controlled pilot study. AB - In this pilot-scale, double-blind, placebo-controlled trial, 30 patients with Helicobacter pylori infection were randomised into three groups prior to their 7 days eradication therapy, to study the effects of probiotic supplement comprising Lactobacillus acidophilus and Bifidobacterium bifidum on the intestinal microflora in response to antibiotic therapy. Group I received the placebo product from day 1 to day 15, Group II received placebo from day 1 to day 7 and probiotics from day 8 to day 15 and Group III received probiotics from day 1 to day 15. Patients provided stool samples for analysis on days 1, 7, 12, 17 and 27. For patients in Groups I and II, significant increases in the facultative anaerobe component of the microflora occurred between days 1 and 7. In Group I, the numbers remained elevated to day 27 but in Group II, the numbers decreased significantly between days 7 and 27 back to the starting levels. In Group III, the facultative anaerobe population remained stable throughout. The total anaerobe numbers increased significantly at day 27 than at day 1 for Group I, were unchanged throughout for Group II and decreased significantly for the patients in Group III between days 1 and 7 before reverting to the starting levels by day 27. From these results, it can be seen that probiotic supplementation modulates the response of the intestinal microflora to the effects of antibiotic therapy. PMID- 15829426 TI - Reconsideration of histopathology and ultrastructural aspects of the human liver in yellow fever. AB - Yellow fever is a re-emerging infectious disease that currently is at risk of urbanization due to the advance of the Aedes aegypti vector. The disease affects about 200,000 individuals annually, mainly in tropical Africa and South America. It causes severe disease involving especially the liver, with lesions characterized by midzonal steatosis, apoptosis and lytic necrosis of the hepatocytes. Quantitative histological and immunohistochemical analysis of 53 human hepatic samples demonstrated apoptosis, steatosis and lytic necrosis of hepatocytes with midzonal pattern. No substantial alterations and reticular network were observed. The inflammatory infiltrate consisted of mononuclear cells and intensity was minimal or moderate, disproportionate to the intense death of the hepatocytes. Hepatic damage in yellow fever resulted mainly from a massive death of hepatocytes due to apoptosis and to a lesser extent due to lytic necrosis. It is recommended that therapeutic regimens for serious cases should include measures to protect against apoptosis. PMID- 15829427 TI - Specific point mutations may not accumulate with aging in the mouse mitochondrial DNA control region. AB - Increasing evidence suggests that mitochondrial function declines during aging in various tissues and in a wide range of organisms. This correlates with an age dependent large accumulation of specific point mutations in the mtDNA control region that was reported recently in human fibroblast and skeletal muscle. However, evaluations of aging-related mtDNA mutations in other model animal systems. In this study, we analyzed mtDNA control regions of brain, skeletal muscle, heart, and other tissues from aged mice, in search of specific point mutations. A 948-bp fragment covering the entire mtDNA control region from various tissues of mice at the age of 25-26 months was sequenced. The sequence analysis was accomplished with a newly developed program Mutation Quantifier, which was able to accurately detect mutations with frequencies as low as 3%. Probably due to the relative shorter life-span, unlike what has been reported in human mtDNA, our results indicated there might be no significant accumulation of specific mutations in mouse mtDNA control region during aging. PMID- 15829428 TI - Cumulative effects of AT1 and AT2 receptor blockade on ischaemia-reperfusion recovery in rat hearts. AB - Though ischaemia/reperfusion injury induces renin-angiotensin systemic (RAS) activation and increased heart angiotensin production, the effects of blockade of the two main angiotensin II receptors, AT1 and AT2, are not definitively established. Using a Langendorff heart preparation, effects of Valsartan 10(-7)M (AT1 receptor blocker), PD 123319 10(-7)M (AT2 receptor blocker) or both in the presence of a controlled concentration of angiotensin II (10(-8)M) in order to reproduce systemic RAS activation were studied in adult male Wistar rat hearts submitted to ischaemia/reperfusion. Ischaemia/reperfusion impaired both systolic and diastolic function through a no-reflow phenomenon. Presence of a controlled concentration of angiotensin in the perfusate, enough to produce a significant AT1-induced vasoconstriction before ischaemia, has no relevant influence on ischaemia/reperfusion injury. Only blockade of both AT1 and AT2 receptors significantly improved recovery from ischaemia; better ventricle function paralleled better perfusion. The results suggest that blockade of angiotensin II receptors is cumulative since blockade of AT1 and AT2 receptors is more effective than blockade of just one of them. PMID- 15829429 TI - Effects of conjugated linoleic acid supplementation on blood lipids and adiposity of rats fed diets rich in saturated versus unsaturated fat. AB - Conjugated linoleic acid (CLA) may decrease adiposity and improve blood lipid profiles under some conditions. The goal of this study was to determine the effects of CLA supplementation on blood lipid profiles and adiposity of rats fed a diet containing a primarily saturated fat versus a diet containing a primarily unsaturated fat. Twenty-eight male Sprague-Dawley rats were randomly assigned to one of four diets containing coconut oil, coconut oil with CLA, corn oil or corn oil with CLA. After 28 days, blood was collected and serum concentrations of total cholesterol (TC), HDL-cholesterol (HDL-C), and triacylglycerols (TG) were assessed. Food intake, body weights, and epididymal fat pads were measured. No significant differences (p>0.05) were noted among groups for amount of food consumed, weight gained, food efficiency ratio or serum TG concentrations. TC concentrations were lower (p<0.05) in the CLA-supplemented rats that were fed coconut oil but not those consuming corn oil. Serum HDL-C was lower (p<0.05) in rats consuming corn oil but was not significantly different (p>0.05) for CLA supplemented groups. Epididymal fat pads weighed significantly more (p<0.05) in the coconut oil fed group compared to the corn oil fed group, but there was no significant difference (p>0.05) between the corn oil and coconut oil + CLA group. Overall, this study suggests that CLA is more beneficial for control of blood lipids and adiposity when supplemented to a diet rich in saturated versus unsaturated fat. PMID- 15829430 TI - Effects of heparin-binding protein (CAP37/azurocidin) in a porcine model of Actinobacillus pleuropneumoniae-induced pneumonia. AB - Heparin-binding protein (HBP; CAP37/azurocidin) is secreted from neutrophil leukocytes early during inflammation and plays a central role in early capillary leakage and extravasation of neutrophils. Furthermore, HBP is chemotactic towards monocytes and lymphocytes and protects against stress-induced apoptosis, e.g. induced by oxygen radicals released during inflammation. Thus, administration of HBP as an adjunct to antibiotics increased survival of mice with peritonitis. In the present study, the effects of recombinant HBP as an adjunct to standard antibiotics were examined in a porcine model of Actinobacillus pleuropneumoniae induced pneumonia. Beneficial and possible adverse effects of HBP were evaluated clinically and pathologically as well as by measurement of clinical chemical variables and markers of inflammation (interleukin-6 and C-reactive protein) and oxidative stress (ascorbic acid and alpha-tocopherol). Pigs receiving HBP (0.55 mgkg-1, n=11) as a 6-hourly subcutaneous infusion starting 1-h post-infection had a faster decrease in rectal temperature compared to control animals receiving a carrier-infusion (n=11), with a significant lower temperature at 32 h post infection (p<0.05). This difference was, however, transient and the temperature curves had a similar course from 38 h and onwards. The transient effect of HBP might be explained by the dosage regimen that was used. No signs of adverse effects of the HBP-infusion were observed. More studies are needed to elucidate of the effects of HBP further and to optimise the dosage regimen for further improvement the efficacy and safety. PMID- 15829431 TI - Histamine-induced vasodilation in the perfused kidney of STZ-diabetic rats: role of EDNO and EDHF. AB - In this study, we have examined the contribution of endothelium-derived nitric oxide (EDNO) and endothelium-derived hyperpolarizing factor (EDHF) to histamine induced endothelium-dependent relaxation in the perfused kidney of rats treated with streptozotocin (STZ) to induce diabetes. Histamine-induced vasodilatation in the perfused kidney preparations of both control and diabetic animals, which was not significantly different. Sodium nitroprusside (SNP)-induced relaxation was also not affected in diabetic and control rats. In order to isolate the EDHF component of histamine-induced vasodilator response, L-NAME (10(-4)M) and indomethacin (10(-6)M) were added to the Krebs' solution throughout the experiment. TBA (0.5 mM) produced a significant reduction in histamine-induced maximal vasodilator response in both preparations from control and diabetic animals, indicating the involvement of K+ channels in mediating this response. Charybdotoxin (0.05 microM) but not glibenclamide (0.1 microM) produced significant reduction in histamine-induced vasodilator responses. To test the contribution of EDNO in mediating histamine-induced vasodilatation, the vascular preparations were perfused with 20 mM K+ -Krebs' solution to inhibit the EDHF component of the response. Under this condition, histamine-induced vasodilator response was not significantly different in both preparations from control and diabetic rats. Pre-treatment with L-NAME (10(-4)M) attenuated histamine-induced vasodilatation. There was a more significant attenuation in histamine-induced vasodilatation in the vascular preparations from diabetic rats. The vasodilator effect of calcium ionophore A23187 was investigated in preparations from control and diabetic rats to examine receptor dysfunction associated with diabetes. A23187 produced dose-dependent vasodilator response in the preparations from both control and diabetic rats. In conclusion, our results indicate that histamine induced vasodilatation in the perfused kidney of the STZ-induced diabetic rats is mediated by the two vasodilator components, namely EDHF and EDNO. The EDHF component was not significantly affected by diabetes. However, histamine-induced vasodilatation mediated by the EDNO component was more significantly reduced in diabetic rats. Results have also indicated that the EDHF component of histamine induced vasodilatation was mediated through Ca2+ -activated K+ channels in perfused kidney preparations from both control and diabetic rats. PMID- 15829432 TI - Differential effect of antioxidant treatment on plasma and tissue paraoxonase activity in hyperleptinemic rats. AB - Recent studies suggest that adipose tissue hormone, leptin, is involved in atherogenesis, especially in obese subjects. Previously, we have demonstrated that experimentally induced hyperleptinemia decreases plasma paraoxonase 1 (PON1) activity. The aim of this study was to investigate whether treatment with synthetic antioxidant, Tempol, modulates the effect of leptin on plasma and tissue PON1 in the rat. Leptin was administered at a dose of 0.25 mgkg-1 s.c. twice daily for 7 days and Tempol was added to the drinking water at a concentration of 2 mM. Leptin reduced plasma PON1 activity toward paraoxon, phenyl acetate and gamma-decanolactone to 71.1, 72.3 and 57.1% of control, respectively. In addition, leptin decreased PON1 activity toward paraoxon in aorta, renal cortex and medulla to 78.6, 49.2 and 48.0% of control, respectively, but had no effect on PON1 in heart, lung and liver. PON1 activity toward phenyl acetate was lower following leptin treatment only in aorta. Leptin increased plasma concentration and urinary excretion of isoprostanes as well as malonyldialdehyde + 4-hydroxyalkenals level in aorta, renal cortex and renal medulla. Coadministration of Tempol prevented leptin-induced oxidative stress and normalized PON1 activity in aorta and kidney. However, Tempol had no effect on plasma PON1 in leptin-treated rats. These data indicate that hyperleptinemia decreases tissue PON1 activity through oxidative stress-dependent mechanism. In contrast, leptin-induced downregulation of plasma PON1 is not mediated by oxidative stress. PMID- 15829433 TI - Pharmacological effects mediated by UDP-glucose that are independent of P2Y14 receptor expression. AB - In transfected cells, the P2Y14 receptor reportedly couples to pertussis toxin sensitive G(i/o)-proteins. However, the functional coupling of endogenously expressed P2Y14 receptors to the inhibition of adenylyl cyclase activity has not been reported. Therefore, the primary aim of this study was to investigate the effects of uridine 5'-diphosphoglucose (UDP-glucose) on forskolin-stimulated cyclic AMP (cAMP) accumulation in two cell lines that reportedly express P2Y14 receptor mRNA, namely human neuroblastoma SH-SY5Y cells and human astrocytoma U373 MG cells. In U373 MG cells, UDP-glucose inhibited forskolin-stimulated cAMP accumulation in a concentration-dependent manner (pEC50=4.5 +/- 0.3). Furthermore, treatment with pertussis toxin abolished the inhibitory effects of UDP-glucose on forskolin-stimulated cAMP accumulation in U373 MG cells. In SH SY5Y cells, UDP-glucose had no significant effect on forskolin-stimulated cAMP accumulation. To confirm the expression of P2Y14 receptor mRNA in U373 MG and SH SY5Y cells, we performed reverse transcriptase polymerase chain reaction (RT-PCR) analysis. However, RT-PCR did not detect the expression of P2Y14 receptor mRNA in SH-SY5Y cells or surprisingly in U373 MG cells. In conclusion, we have shown that although UDP-glucose inhibits forskolin-stimulated cAMP accumulation in human U373 MG astrocytoma cells, we did not detect P2Y14 receptor mRNA in these cells. These results would suggest that the effects of UDP-glucose in U373 MG cells are independent of P2Y14 receptor expression. Thus, results obtained with UDP-glucose should be interpreted with caution, since they clearly may not necessarily reflect the involvement of the P2Y14 receptor. PMID- 15829434 TI - Amentoflavone inhibits the induction of nitric oxide synthase by inhibiting NF kappaB activation in macrophages. AB - Amentoflavone is a bi-flavonoid compound with anti-fungal and anti-inflammatory activities. We isolated amentoflavone from Selaginella tamariscina (Selaginellaceae) and studied its effects on nuclear factor-kappaB (NF-kappaB) mediated inducible nitric oxide synthase (iNOS) gene expression in RAW 264.7 cells. Amentoflavone inhibited the production of nitric oxide in a concentration dependent manner and also blocked the lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase (iNOS). To clarify the mechanistic basis for its inhibition of iNOS induction, we examined the effect of amentoflavone on the transactivation of iNOS gene by luciferase reporter activity using -1.59 kb flanking region. Amentoflavone potently suppressed the reporter gene activity. The LPS-induced activation of NF-kappaB was also found to be significantly blocked by amentoflavone, but AP-1 activation was unaffected. Furthermore, the nuclear translocation of p65 by LPS was inhibited by amentoflavone. NF-kappaB activation is controlled by the phosphorylation and subsequent degradation of I kappaBalpha, and the cytosolic degradation of I-kappaBalpha was found to be inhibited by amentoflavone. These findings suggest that the inhibition of LPS induced NO formation by amentoflavone is due to its inhibition of NF-kappaB by blocking I-kappaBalpha degradation, which may be the mechanistic basis of the anti-inflammatory effects of amentoflavone. PMID- 15829435 TI - Effects of SNP, GLU and GABA on the neuronal activity of striatum nucleus in rats. AB - This study investigated the activity of nitric oxide (NO) in the striatum (STR) for a further comprehension of the pathogenesis of Parkinson's disease (PD). Microiontophoresis was used to observe the effects of sodium nitroprusside (SNP), L-glutamic acid (GLU) and gamma-aminobutyric acid (GABA) on STR neurons' firing rates. It was observed that 77.27% (51/66) of the tested STR neurons were excited by SNP. This excitatory effect could be antagonized by the NO synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME). During the microiontophoresis of GLU, the excitatory firing of STR neurons was also attenuated by addition of L-NAME while SNP application could enhance the excitation of the neurons. On the other hand, in the presence of GABA, SNP still excited the tested STR neurons. These results demonstrated that NOergic, GLUergic and GABAergic co-existed in the same STR neurons. NOergic and GLUergic were excitatory whereas GABAergic was inhibitory on the firing activity in STR neurons. PMID- 15829436 TI - Down-regulation of cyclooxygenase-2 and telomerase activity by beta-lapachone in human prostate carcinoma cells. AB - Beta-lapachone, the product of a tree Tabebuia avellanedae from South America, is known to exhibit various pharmacologic properties, the mechanisms of which are poorly understood. In the present study, we investigated further possible mechanisms by which beta-lapachone exerts its anti-proliferative action in cultured human prostate carcinoma DU145 cells. Exposure of DU145 cells to beta lapachone resulted in growth inhibition and induction of apoptosis in a dose dependent manner as measured by MTT assay, fluorescent microscopy, and flow cytometry analysis. The increase in apoptosis was associated with a dose dependent up-regulation in pro-apoptotic Bax expression, down-regulation of anti apoptotic Bcl-2, and proteolytic activation of caspase-3 protease. We found beta lapachone decreased the levels of cyclooxygenase (COX)-2 mRNA and protein expression without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin E2 (PGE2) synthesis. Furthermore, beta-lapachone treatment markedly inhibited the activity of telomerase in a dose dependent fashion. Additionally, the expression of human telomerase reverse transcriptase (hTERT), a main determinant of the telomerase enzymatic activity, was progressively down-regulated by beta-lapachone treatment. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of beta-lapachone. PMID- 15829437 TI - Expression and activity of CYP2E1 in circulating lymphocytes are not altered in diabetic individuals. AB - Cytochrome P4502E1 (CYP2E1) plays an important role in ROS production thus favouring accelerated membrane lipid peroxidation. This isoform is strongly expressed in the liver but it can be also found in lymphocytes. As such, lymphocyte may provide a non-invasive accessible pool for screening CYP2E1 expression in man. We have, therefore, analysed CYP2E1 expression and activity in lymphocyte microsomes from 12 healthy controls, 11 type 1 and 12 type 2 diabetic subjects by using Western blot and enzymatic activities. Immunoblotting did not show difference among CYP2E1 protein bands in controls, type 1 and type 2 diabetics. To assess CYP2E1 activity we used the 7-ethoxy-4 trifluoromethylcoumarin (7-EFC), as a fluorescent substrate. The rate of deethylation of 7-EFC from controls did not differ from type 1 and type 2 diabetic subjects. The lack of any difference in CYP2E1 activity also was confirmed by the NADPH-dependent microsomal lipid peroxidation CCL4-induced assay showing similar peroxidation rates among controls and diabetic subjects. The results show that CYP2E1 expression/activity in lymphocytes is not enhanced in diabetes. PMID- 15829438 TI - The contracting and relaxing responses of human internal mammary artery grafts harvested by two different methods. AB - Perioperative spasm of internal mammary artery is a common experience in coronary artery bypass grafting. Many techniques were described of harvesting the internal mammary artery to prevent vasospasm. We investigated the comparison of the contracting and relaxing responses of human internal mammary artery grafts harvested by two different methods. Patients were divided into two groups depending on the harvesting technique. In the first and second groups arteries were harvested by classical and carbon dioxide insufflation techniques, respectively. In both groups, endothelial function of arteries was assessed by precontracting the rings with phenylephrine (10(-5)M) and dilatating them by cumulative acetylcholine (10(-8) to 10(-5)M) concentrations. Cumulative concentration-response curves for phenylephrine (10(-8) to 10(-4)M), noradrenaline (10(-9) to 10(-4)M), and 5-hydroxytryptamine (10(-9) to 10(-4)M) were obtained in all groups. Endothelial integrity of arteries were histopathologically evaluated. In both groups, acetylcholine caused concentration dependent relaxations in rings precontracted with phenylephrine (10(-5)M). In arteries harvested by carbon dioxide insufflation technique, acetylcholine caused significantly higher relaxations compared to the rings obtained by classical technique (p<0.05). In all rings of study groups, phenylephrine, noradrenaline and 5-hydroxytryptamine caused concentration-dependent contractions. There was not any significant difference in concentration-dependent responses of these contracting pharmacological agents between the groups. Histopathological evaluation revealed no major arterial damage in both groups. Carbon dioxide insufflation technique does seem not only to protect the integrity of the endothelium and the whole vessel, but also prevent the possible vasospasm of the internal mammary artery segments. PMID- 15829439 TI - Study of urinary 6 beta-hydroxycortisol/cortisol ratio in spot urine sample as a biomarker of 3A4 enzyme activity in healthy and epileptic subjects of Egyptian population. AB - The ratio of urinary 6 beta-hydroxycortisol/cortisol (6 beta-OHC/FC) in morning spot urine samples collected from 8:00 a.m. to 12:00 p.m. was studied using ELIZA kits (Stabiligen) in a group of healthy adult Egyptians (control group) of both sex (n=65, age range: 16-48 years). The frequency distribution of urinary 6 beta OHC/FC ratio was widely distributed among subjects with higher values in males in comparison to females. No bimodality in either sex was observed. Another group of adult epileptic patients (n=16) was studied for the influence of chronic carbamazepine antiepileptic drug administration on urinary 6 beta-OHC/FC ratio in spot urine samples. The induction property of carbamazepine on CYP3A4 was observed through significant increase (p=0.01) in 6 beta-OHC/FC ratio among epileptic patients in comparison with control subjects. In conclusion, the frequency distribution of urinary 6 beta-OHC/FC ratio among Egyptians shows sexual dimorphism. Also, measurement of urinary 6 beta-OHC/FC ratio provides a simple non-invasive method to monitor CYP3A4 enzyme induction during administration of carbamazepine antiepileptic drug. PMID- 15829440 TI - Inhibition of poly(ADP-ribose) polymerase prevents vascular hyporesponsiveness induced by lipopolysaccharide in isolated rat aorta. AB - Recent studies clearly show that there is a relationship between endotoxemia and impaired vascular responsiveness. The aim of this study was to investigate whether treatment with the new potent PARP inhibitor PJ34 could prevent the vascular hyporesponsiveness induced by lipopolysaccharide (LPS). Endotoxemia was induced in rats by LPS injection (20 mgkg-1, i.p.). Administration of LPS caused a decrease in mean blood pressure and an increase in heart rate. In endothelium denuded rings of thoracic aorta from untreated rats, contractile responses to KCl and phenylephrine decreased after LPS injection. Furthermore, there was a significant loss of endothelium-dependent vasodilatation in response to acetylcholine in LPS-treated rats. The animals pretreated with PJ34 (10 mgkg-1, i.p., 30 min before LPS injection), the effect of LPS on vascular responsiveness was lower than the untreated ones. Pretreating the animals with PJ34 before the LPS challenge prevented the decline in mean blood pressure. However, this did not result in significant changes to the heart rate. The inhibitory effect of LPS treatment on both KCl- and phenylephrine-induced contraction responses was significantly antagonized by PJ34. Additionally, pretreatment of the rats with PJ34 attenuated the LPS-induced endothelial dysfunction in endothelium-intact aorta rings. This study demonstrates that PARP activation in the vascular system is an important contributory factor to the impaired vascular responsiveness associated with endotoxic shock. Hence, the pharmacological inhibition of PARP pathway might be an effective intervention to prevent endotoxin-induced vascular hyporesponsiveness. PMID- 15829441 TI - Interaction of topiramate with glycine receptor channels. AB - Glycine receptor channels are pentameric ligand-gated ion channels that respond to the application of inhibitory neurotransmitters by opening of a chloride selective central pore. Topiramate (TPM) is a broad-spectrum antiepileptic drug used as add-on or monotherapy for focal seizures. In the present study the interaction of TPM with glycine receptor channels was studied on outside-out patches from HEK293 cells expressing alpha1beta glycine receptor channels. The patch clamp techniques combined with ultra fast solution exchange enabled us to investigate the kinetics of receptor channels in presence of TPM. Our study showed no agonistic or potentiating effect for TPM on glycine receptor channels. However, in presence of 1 mM glycine + 1 mM TPM, the desensitization got faster and the peak current amplitude decreased. After the end of glycine + TPM pulses, off-currents occurred, suggestive for a specific channel block mechanism. PMID- 15829442 TI - The double burden of and negative spillover between paid and domestic work: associations with health among men and women. AB - The objective of the study was to determine whether the double burden of and negative spillover between domestic and full-time paid work are associated with an increase in health problems. Survey responses were linked with sickness absence records in a cross-sectional study of 6442 full-time municipal employees. Women and men experiencing severe work-family spillover had a 1.5-1.6 (95% confidence intervals 1.1 to 2.0) times higher rate of sickness absence than those with no such experience. The corresponding odds ratios for psychological distress and suboptimal self-assessed health varied between 3.6 and 6.5 (2.3 to 11.0). Among the women, severe family-work spillover increased the risk of psychological distress and suboptimal health [odds ratios 2.0 (1.4 to 2.9) and 1.6 (1.1 to 2.3), respectively], and accumulated sole responsibilities were associated with a 1.5 (1.1 to 2.1) times higher odds ratio for psychological distress. Long domestic work hours (>50/week) were associated with a 1.5 (1.1 to 2.0) times higher rate of sickness absence among the men, but there was no such increase among the women. We conclude that negative work-family spillover especially is associated with health problems among both women and men, and negative family work spillover is related to a poorer health status among women. PMID- 15829443 TI - Comparing women of differing sexual orientations using population-based sampling. AB - OBJECTIVES: Area probability sampling was used to conduct a women's health survey in Boston, MA. Sexual minority women (SMW) and heterosexual adult women were compared on a variety of health-related measures. METHODS: SMW-rich census tracts were identified and mapped onto zip code boundaries. Eligible respondents were women 18 and older who lived within the defined area, who were able to complete a personal interview or self-administered questionnaire in English. Differences in significant health-related outcomes by sexual orientation were examined. RESULTS: SMW and heterosexual women differed on access to health care and utilization of screening tests. There were no significant differences in smoking rates, eating less calories or fat, and intentions to follow mammography recommendations. CONCLUSIONS: In certain respects, study results are congruent with previous non probability surveys, while in others the results are different. It is likely that real differences exist in some health-related variables by sexual orientation category. PMID- 15829444 TI - Female methamphetamine users: social characteristics and sexual risk behavior. AB - The primary objective of this research was to expand our knowledge regarding the personal and social characteristics of female methamphetamine (meth) users, their motivations for using meth, patterns of meth use, medical and social problems associated with meth use, and the relationship between meth use and sexual risk behaviors. The sample consisted of 98 HIV-negative, heterosexually-identified, meth-using females residing in San Diego, California. Female meth users were characterized by personal and social disadvantage, high rates of psychiatric symptomatology, and high levels of sexual risk behavior, including multiple partners, risky partner types (e.g., anonymous sex partners), and high rates of unprotected vaginal and oral sex. Meth use was also associated with the subjective positive experience of sex. These finding suggest that behavioral interventions should be tailored to the social characteristics of female meth users, and program content should reflect the intertwining of women's sexual experience and meth use. PMID- 15829445 TI - Religious beliefs, faith community involvement and depression: a study of rural, low-income mothers. AB - The current study investigated the connection between religion and mental health of 131 rural, low-income mothers. Two dimensions of religion, beliefs and faith community involvement, were included and depression was assessed by the CES-D. The sample consisted of mothers who participated in Wave 2 of a multi-state research project. As hypothesized, both religious beliefs and faith community involvement were negatively related to depressive symptoms indicating that mothers with stronger religious beliefs and more involvement in religious activities may experience less depressive symptoms. The results of the current study confirm previous work and support a multifaceted view of religion. PMID- 15829446 TI - Moving beyond disclosure: women's perspectives on barriers and motivators to seeking assistance for intimate partner violence. AB - To investigate women's perceptions of motivators and barriers to seeking help or accessing intimate partner violence (IPV), services six focus groups were conducted in rural and urban settings in North Carolina between June and August of 2002. Coding and theme analysis were used to summarize themes among the 67 focus group participants. The majority of participants were African-American (87%). Participants reported three main categories of motivators: gaining knowledge; reaching an emotional or physical breaking point; and growing concern about children's safety. Participants reported six main categories of barriers: pressure not to talk about, or address IPV; failure to recognize events as IPV, or that IPV was wrong; self-doubt and low self-esteem; fear of losses; fear of perpetrator; or desire to protect the perpetrator. This study documents the difficulties that women face accessing or using services related to IPV. We need to address perceived barriers and better use the opportunity when women experience motivation to seek help and access services. PMID- 15829447 TI - Generations Exercising Together to Improve Fitness (GET FIT): a pilot study designed to increase physical activity and improve health-related fitness in three generations of women. AB - A 6-month home-based (HB) physical activity program was compared to a control (CTL) condition in terms of effect on physical activity and health-related fitness in three generations of women (daughter/ mother/maternal grandmother). Volunteers were randomly assigned to a HB or CTL condition. HB participants (n = 28) were asked to participate in lifestyle, aerobic, muscular strength, and flexibility activities at least 3 times per week and they completed 73% of the recommended PA bouts. CTL condition participants (n = 9) were asked to continue their usual pattern of physical activity. Changes in physical activity were measured pre-and post-intervention using the Physical Best questionnaire and pedometer step counts (3-day average). Changes in health-related fitness were assessed using Fitnessgram tests. Group x Time interactions were significant for changes in participation in flexibility activity (d/wk) and steps/day, indicating that the HB group experienced significant positive changes in the expected direction (+305% and +37%, respectively), while the CTL group regressed (-15% and -13%, respectively). The G x T interaction for mile time was significant, although not in the expected direction (CTL group < by 14% and HB group < by 5%). Findings should be interpreted with caution due to several limitations of the study, but several suggestions are made for more effectively studying this topic in the future. PMID- 15829448 TI - Domestic violence against 116 Turkish housewives: a field study. AB - In order to clarify the factors associated with domestic violence against married women and to learn the attitude of women toward violence and toward the inquiry of health personnel about domestic violence, a cross-sectional survey was conducted on 116 married Turkish women. The prevalence of domestic physical violence was found to be 41.4%, emotional violence 25.9%, sexual violence 8.6%, and controlling behavior 77.6%. Physical violence experienced at any time in their life was 50.9%. Rate of physical violence was found related with lower formal education of both women and husbands and their place of birth. The opinion of "husbands must not beat their wives" was more common among women with formal education. Eighty-two percent of women indicated that they would be willing to answer questions regarding domestic violence by health personnel. In conclusion, cultural background and formal education of either woman or man influence the likelihood of imposing or tolerating violent behavior at home. Inquiry about violence may be a routine in women's health care practice, and it is welcome by most women. PMID- 15829449 TI - Exploring Chinese women's cultural beliefs and behaviours regarding the practice of "doing the month". AB - It is well recognized that individual health behavior is embedded in cultural patterns of exchange. This study sought to identify Chinese women's cultural beliefs and behaviors related "doing the month" and suggest how these are modified in the light of contemporary realities and experiences within the context of rapid social change. Ethnographic interview data was gathered by means of a telephone interviews (postnatally) with 100 eligible primiparous women who had attended hospital based antenatal programmes. The analysis highlighted a range of issues for these predominantly middle class women which included special postpartum dietary beliefs and behaviours including the avoidance of hot and cold food, the restorative powers of food, wind and water prohibitions, food prescriptions and prohibitions during breast feeding, the cultural imperative of timing and contemporary autonomy of women versus conforming to female centered kinship obligations. It is concluded that health professionals working with Chinese women need further awareness of the cultural significance of adherence to post natal practices and their fundamental links to the health status of post partum women, and family relationships. PMID- 15829450 TI - The story behind the story of collaborative networks -- relationships do matter! AB - This study reports data about the real story behind the current trend of mandated interorganizational collaboration of health and human service agencies. By means of qualitative design (N-22), public health managers were interviewed about the extent and nature of their collaborative efforts in the Healthy Babies, Healthy Children (HBHC) Program in Ontario, Canada. Using a conceptual framework of resource exchange theory, this study found that relational processes specifically: (a) previous relationships with other agencies and (b) interpersonal relations namely: informality, local community, open communication and resolving conflicts were the reasons for successful collaborations. Implications are directed toward: health and social planners, administrators, board members, funding bodies and policy-makers. The study offers new knowledge about a subject which has received minimal attention in the literature. PMID- 15829451 TI - Drug use and harm reduction policy: alternative perspectives on the California proposal. AB - On October, 12, 2003, then California Governor Gray Davis vetoed legislation which would have created a trial period for evaluating nonprescription pharmacy access to needles and syringes for adults in California. An analysis of California Senate Bill 774 provides a case study to examine the struggle for clean syringe and needle access as a component of disease prevention along with the community fear of tacit approval for illicit drug use. Reflecting upon the empirically-supported research and community concerns for this specific type of harm reduction policy is an important step towards forming common political ground. This article underscores the relevance of syringe deregulation and its relevance as an ethical and political issue for social workers. PMID- 15829452 TI - What providers and medicaid policymakers need to know about barriers to employment for people with disabilities. AB - Medicaid Buy-Ins provide a new and exciting opportunity for people with disabilities to engage in meaningful employment while maintaining Medicaid coverage. Through interviews with participants in the Kansas Medicaid Buy-In, we examined perceived external influences on the decision to acquire or increase employment by people with disabilities. Two major external barriers were identified. First, physicians, therapists and case workers had frequently discouraged participants from getting jobs or increasing employment levels. Difficulty accessing adequate and consistent medical care and/or medications through Medicaid was also an issue in preventing participants from being able to acquire or increase employment. PMID- 15829453 TI - The Poverty and Housing Scale: report on a pilot study. AB - The measurement of beliefs and attitudes on poverty and housing is important to researchers and social workers interested in examining the role that belief structures have on the development of policy and programs in these areas. This article reports pilot study findings of a new scale, The Poverty and Housing Scale (PHS), that measures this concept and evaluates its psychometric properties. Preliminary reliability was in the very good range. Examinations of content and face validity provided support of the instrument as a valid measure of beliefs and attitudes on poverty and housing. The factor analysis emerged a one factor, 13-item scale. Unlike other related scales, the PHS attempts to link the social factor of poverty and housing together. Theoretical and methodological strengths and weaknesses are considered and the implications for social work practice are discussed. The authors provide recommendations for additional testing of the instrument. PMID- 15829454 TI - A comparison snapshot of states' small group market reform on access and enhancing valued plan features, 1999. AB - Small group health insurance statutes of 48 states and the District of Columbia, as of 1999, were reviewed. Reform provisions judged to have some relevance for the market are catalogued and total 74 distinct regulations. Judgment of market relevance was based on actuarial experience at a leading health insurance company. The regulations are categorized under: (1) Access improvement; (2) Pricing and Rating Reforms; (3) Improving stability of coverage; and (4) Improving valued features of plans. The nuances and variety of these regulations, adopted in various combinations by the states, are discussed. The complexity of the reform scenario suggests the need for impact studies that take into account the totality of reform. Past studies have evaluated the impact of selected major reforms in isolation, and, thus, have been inadequate to provide definitive conclusions on the reforms' impact. PMID- 15829455 TI - PDA power at the bedside. AB - George Washington University's Himmelfarb Health Sciences Library was investigating adding PDA resources to the collection when a medical education grant was received. Funds were used for a pilot project to compare PDA use by third-year medical students (MSIII), fourth-year medical students (MSIV), second year physician assistant (PAII) students, and residents in the clinical setting. The PDA Committee evaluated various titles by the following criteria: both Palm and PocketPC operating systems compatibility, ease of use, depth of information, product cost, and a wide target audience. Griffith's 5 Minute Clinical Consult by Skyscape was selected. Library registration was required to receive the software and PDA downloading instructions. A short, Web-based questionnaire was developed to evaluate the product's use in the clinical setting. A low response rate of 74 questionnaires (23% of participants) was received and analyzed in March 2004. As additional funding becomes available, PDA software will be seriously considered as additions to the collection. PMID- 15829456 TI - Questions asked at the virtual and physical health sciences reference desk: how do they compare and what do they tell us? AB - The questions asked at the traditional reference desk are decreasing while questions asked at the virtual reference desk are on the rise. Over a one-month period, the types of reference questions asked at an academic health sciences library were coded. This paper examines and compares the types of questions asked at the current day reference desk versus the virtual reference desk. This paper also reviews past literature examining the types of questions asked via virtual reference and the traditional reference. PMID- 15829457 TI - Client value models provide a framework for rational library planning (or, phrasing the answer in the form of a question). AB - Libraries often do not know how clients value their product/ service offerings. Yet at a time when the mounting costs for library support are increasingly difficult to justify to the parent institution, the library's ability to gauge the value of its offerings to clients has never been more critical. Client Value Models (CVMs) establish a common definition of value elements-or a "value vocabulary"-for libraries and their clients, thereby providing a basis upon which to make rational planning decisions regarding product/service acquisition and development. The CVM concept is borrowed from business and industry, but its application has a natural fit in libraries. This article offers a theoretical consideration and practical illustration of CVM application in libraries. PMID- 15829458 TI - Extending the reference desk experience: Information Resources for the Nursing Care Plan. AB - Both nursing and library science literature discuss the information needs and information-seeking behavior of nurses. Some research has focused on information needs of nursing students, but little research is available regarding information seeking behavior of nursing students, specifically baccalaureate nursing students. Only a few key references were identified addressing nursing students' information seeking with regard to nurse care planning. At the University of Medicine and Dentistry of New Jersey, George F. Smith Library, a wide range of nursing students from those in traditional BSN programs, to students in accelerated BSN programs, to masters' nursing students of all types visit the reference desk asking direct and indirect questions involving the nursing care plan. This article highlights repeated categories of information sought by nursing students at the University of Medicine and Dentistry of New Jersey, George F. Smith Library via both formal and informal reference interviews. Information sought most frequently was categorized and formed the basis for a new library workshop entitled "Information Resources for the Nursing Care Plan." PMID- 15829459 TI - What is BioOne? AB - BioOne is a Web-based aggregation of full-text, high-impact bioscience research journals. Most of its titles are published by small societies or non-commercial publishers and have not been previously available in electronic format. This column describes the BioOne database and gives some basic information about the best ways to search its content. PMID- 15829460 TI - Evidenced-based nursing resources. AB - Evidence-based medicine and, more generally, evidence-based practice (EBP), is the integration of the best available patient care evidence and clinical expertise-guided by and sensitized to patient values-into a patient-oriented decision-making process. It is increasingly clear that all health professions neither equally value nor draw upon the same pool of evidence. Although there are areas of overlap between evidence-based nursing and medicine, for example, there are important differences as well. Accordingly, this selective list of Web-based evidence-based nursing (EBN) resources attempts to capture the broader scope and practice of nursing in terms of its focus, subject matter, and research methods. PMID- 15829461 TI - Author! author!: creating a digital archive of publications in a hospital library setting. AB - Baptist Hospital of Miami has been honoring its staff authors annually during National Library Week since 1979, at the time the library was relocated. Upon "doing the math" and realizing that twenty-five years had passed, a special event was planned to celebrate the occasion in 2004. A merger of four hospitals in 1995 to form Baptist Health South Florida, and an addition of a fifth hospital in 2003 added into the complexity of these publications. Organizing the event led to the conclusion that there had to be a "better way" to manage the publication archive. This paper will include a look back at the event's past, present efforts to develop an archival database, and future plans to make articles available electronically to users, copyright permitting. PMID- 15829462 TI - Defining and assessing medical informatics competencies. AB - As academic health sciences libraries assume larger roles in informatics instruction within medical school curricula, librarians are challenged to develop useful and accurate measures for assessing the effectiveness of instructional approaches. The need for this evaluation has intensified as medical schools increase their emphasis on integration of curriculum content and shift to competency-based education and assessment of medical students. This paper reports on a pilot project developed at Dahlgren Memorial Library, Georgetown University Medical Center, for two courses using an instructional intervention and tailored assignment for assessing student competencies. PMID- 15829468 TI - Evaluation of valuation of toxicity profile of an alkaloidal fraction of the stem bark of Picralima nitida (fam. Apocynacaes). AB - Dermal and acute toxicity evaluation of the basic alkaloidal fraction of the stem bark of Picralima nitida, which has been shown to have pronounced activity against causative organisms of dermatomycosis in man, was carried out in animals. Acute intraperitoneal toxicity tests showed a dose-dependent toxicity. There was inflammation and necrosis of liver hepatocytes accompanied by reduction in neutrophilic count and a corresponding increase in lymphocytic count. There was no sign of reddening or irritation when applied into the eye conjunctiva. Dermal tests also showed that the fraction caused no sensitization, inflammation or death in the animal models used. PMID- 15829470 TI - Boswellia: an evidence-based systematic review by the Natural Standard Research Collaboration. AB - An evidence-based systematic review including written and statistical analysis of scientific literature, expert opinion, folkloric precedent, history, pharmacology, kinetics/dynamics, interactions, adverse effects, toxicology, and dosing. PMID- 15829469 TI - Analysis of responses to St. John's Wort in the feline pulmonary vascular bed. AB - OBJECTIVE: To test the hypothesis that St. John's wort induces a depressor response in the feline pulmonary vascular bed and identify the pathways involved in the mediation or modulation of these effects. DESIGN: Prospective vehicle controlled study. SETTING: University research laboratory. SUBJECTS: Intact chest preparation; adult mongrel cats. INTERVENTIONS: In separate experiments, the effects of L-N5-(1-Iminoethyl) ornithine hydrochloride (L-NIO), a nitric oxide synthase inhibitor, glibenclamide, an ATP-sensitive K+ channel blocker, meclofenamate, a non-selective cyclo-oxygenase (COX) inhibitor, nicardipine, a calcium channel blocker, bicuculline, a GABAA receptor antagonist, and saclofen, a GABAB antagonist, were investigated on pulmonary arterial responses of St. John's wort (SJW), pinacidil, an ATP-sensitive K+ channel activator, bradykinin, an inducer of nitric oxide synthase, 3-aminopropyl (methyl) phosphinic acid, hydrochloride (SKF-97541), a GABAB receptor agonist and muscimol, a GABAA receptor agonist. MEASUREMENTS AND MAIN RESULTS: Lobar arterial perfusion pressure and systemic pressure were continuously monitored, electronically averaged and permanently recorded. Under elevated tone conditions in the isolated left lower lobe vascular bed of the cat, SJW induced a dose-dependent vasodepressor response that was not significantly altered after administration of L-NIO, glibenclamide, meclofenamate or saclofen. Responses to SJW were significantly reduced after administration of either nicardipine or bicuculline. When the calcium channel blocker nicardipine was administered in addition to the GABA blocker bicuculline, there was near complete attenuation of the SJW-induced vasodepressor responses. CONCLUSIONS: The results of the present study suggest that SJW has potent vasodepressor activity in the pulmonary vascular bed of the cat and that this response is mediated or modulated by both a calcium channel and GABA receptor sensitive pathway. PMID- 15829471 TI - What is eHealth (3): a systematic review of published definitions. AB - CONTEXT: The term eHealth is widely used by many individuals, academic institutions, professional bodies, and funding organizations. It has become an accepted neologism despite the lack of an agreed-upon clear or precise definition. We believe that communication among the many individuals and organizations that use the term could be improved by comprehensive data about the range of meanings encompassed by the term. OBJECTIVE: To report the results of a systematic review of published, suggested, or proposed definitions of eHealth. DATA SOURCES: Using the search query string "eHealth" OR "e-Health" OR "electronic health", we searched the following databases: Medline and Premedline (1966-June 2004), EMBASE (1980-May 2004), International Pharmaceutical Abstracts (1970-May 2004), Web of Science (all years), Information Sciences Abstracts (1966 May 2004), Library Information Sciences Abstracts (1969-May 2004), and Wilson Business Abstracts (1982-March 2004). In addition, we searched dictionaries and an Internet search engine. STUDY SELECTION: We included any source published in either print format or on the Internet, available in English, and containing text that defines or attempts to define eHealth in explicit terms. Two of us independently reviewed titles and abstracts of citations identified in the bibliographic databases and Internet search, reaching consensus on relevance by discussion. DATA EXTRACTION: We retrieved relevant reports, articles, references, letters, and websites containing definitions of eHealth. Two of us qualitatively analyzed the definitions and coded them for content, emerging themes, patterns, and novel ideas. DATA SYNTHESIS: The 51 unique definitions that we retrieved showed a wide range of themes, but no clear consensus about the meaning of the term eHealth. We identified 2 universal themes (health and technology) and 6 less general (commerce, activities, stakeholders, outcomes, place, and perspectives). CONCLUSIONS: The widespread use of the term eHealth suggests that it is an important concept, and that there is a tacit understanding of its meaning. This compendium of proposed definitions may improve communication among the many individuals and organizations that use the term. PMID- 15829472 TI - The role of an online community for people with a rare disease: content analysis of messages posted on a primary biliary cirrhosis mailinglist. AB - BACKGROUND: This study focuses on the role of an Internet-based group for people who have an autoimmune liver disease, primary biliary cirrhosis. Primary biliary cirrhosis is a relatively rare disease, affects primarily women in their 40's and older, and is not well understood. The PBCers Organization (PBC stands for primary biliary cirrhosis) provides electronic mailinglists (listservs) and informational resources for those with primary biliary cirrhosis. OBJECTIVES: (1) to identify the issues of greatest importance to those posting to the listserv, specifically the relative importance of biomedical, socioemotional, and organizational/systems messages; (2) to compare frequency and content of posts by people at different stages of disease; (3) to identify how people with primary biliary cirrhosis represent the psychosocial challenges and dilemmas (role and identity change, uncertainty, and stigma) identified in the social-scientific literature as key elements of the experience of chronic disease. METHODS: The paper is based on content analysis of messages posted during two months to the Daily Digest listserv for people who have primary biliary cirrhosis. To analyze the posts, we developed a coding system with three major categories--biomedical, socioemotional, and systems/organizations--and 12 codes in each category. RESULTS: A total of 275 people posted 710 messages. Of the 250 people for whom information on gender was available, 239 (95.6%) were women and 11 (4.4%) were men. Analysis of 710 messages posted to the listserv revealed a predominance of requests for and reports of biomedical information, such as health care providers (32.7%), medications (30.9%), tests and procedures (25.8%), and symptoms (25.7%), combined with very frequent expressions of emotional support. The most frequent single topics were peer support (included in 40.6% of all posts) and positive emotions (25.3%). Posters who reported fewer years since diagnosis were more likely to be seeking biomedical information than those who were further in time from their diagnosis (r= -.241, P<.001, n=313). Those in later stages posted an average of 3.87 messages, compared to an average of 2.64 for people in earlier stages (t= 1.786, P=.08, n=90), which is different from what we expected. No relation between years since diagnosis or age and number of messages was found. Contrary to our expectations, the topics reflecting issues of role change/identity (2.9%), stigma (0.7%), and thoughts about the future (3.9%), all identified in social-scientific literature as key concerns for people with chronic illness, appeared infrequently in this set of messages. CONCLUSIONS: Messages exchanged on this particular mailing list have a biomedical, rather than socioemotional or organizational, emphasis. The Internet offers a highly valued opportunity for those with rare diseases to connect with, learn from, and provide support to others having similar experiences. Research that compares those with primary biliary cirrhosis, who are involved in an Internet support group and those who are not, would be an important next step to better understanding the role of the Internet among patients with chronic liver disease and the implications of it in the course of their illness. PMID- 15829473 TI - The law of attrition. AB - In an ongoing effort of this Journal to develop and further the theories, models, and best practices around eHealth research, this paper argues for the need for a "science of attrition", that is, a need to develop models for discontinuation of eHealth applications and the related phenomenon of participants dropping out of eHealth trials. What I call "law of attrition" here is the observation that in any eHealth trial a substantial proportion of users drop out before completion or stop using the application. This feature of eHealth trials is a distinct characteristic compared to, for example, drug trials. The traditional clinical trial and evidence-based medicine paradigm stipulates that high dropout rates make trials less believable. Consequently eHealth researchers tend to gloss over high dropout rates, or not to publish their study results at all, as they see their studies as failures. However, for many eHealth trials, in particular those conducted on the Internet and in particular with self-help applications, high dropout rates may be a natural and typical feature. Usage metrics and determinants of attrition should be highlighted, measured, analyzed, and discussed. This also includes analyzing and reporting the characteristics of the subpopulation for which the application eventually "works", ie, those who stay in the trial and use it. For the question of what works and what does not, such attrition measures are as important to report as pure efficacy measures from intention-to-treat (ITT) analyses. In cases of high dropout rates efficacy measures underestimate the impact of an application on a population which continues to use it. Methods of analyzing attrition curves can be drawn from survival analysis methods, eg, the Kaplan-Meier analysis and proportional hazards regression analysis (Cox model). Measures to be reported include the relative risk of dropping out or of stopping the use of an application, as well as a "usage half-life", and prediction models reporting demographic usage discontinuation in a population. Differential dropout or usage rates between two interventions could be a standard metric for the "usability efficacy" of a system. A "run-in and withdrawal" trial design is suggested as a methodological innovation for Internet-based trials with a high number of initial dropouts/nonusers and a stable group of hardcore users. PMID- 15829474 TI - Comparing the efficacy of two Internet-based, computer-tailored smoking cessation programs: a randomized trial. AB - BACKGROUND: Online computer-tailored smoking cessation programs have not yet been compared directly. OBJECTIVE: To compare the efficacy of two Internet-based, computer-tailored smoking cessation programs. METHODS: Randomized controlled trial conducted in 2003-2004. Visitors to a smoking cessation website were randomly assigned to either an original online, interactive smoking cessation program or to a modified program. Both programs consisted of tailored, personalized counseling letters based on participants' characteristics, followed by monthly email reminders. The original program was based on psychological and addiction theory, and on preliminary research conducted in the same population. The modified program was shorter and contained more information on nicotine replacement therapy and nicotine dependence, and less information on health risks and coping strategies. In both programs, 1 month and 2 months after entering the study, participants were invited by email to answer the same tailoring questionnaire again in order to receive a second counseling letter. Participants in both programs obtained, on average, 1.2 feedback counseling letters over 2.5 months, and 84% received only 1 feedback letter. The outcome was self-reported smoking abstinence (no puff of tobacco in the previous 7 days), assessed 2.5 months after entry in the program. We report results from intention-to-treat (ITT) analyses, where all non-respondents at follow-up were counted as smokers. RESULTS: The baseline questionnaire was answered by a total of 11969 current (74%) and former (26%) smokers, and the follow-up survey by 4237 people (35%). In an ITT analysis, abstinence rates in baseline current smokers were respectively 10.9% and 8.9% (odds ratio [OR]=1.24, 95% confidence interval [CI]1.08-1.43, P=.003) in the original and modified programs, and 25.2% and 15.7% (OR=1.81, CI 1.51-2.16, P<.001) in baseline former smokers. While we found statistically significant differences in quit rates in smokers in the contemplation stage favoring the original program (OR=1.54, CI 1.18-2.02, P=.002), no between-group differences in quit rates were observed in smokers in the precontemplation (OR=1.07, CI 0.36-3.14, P=.91) and preparation (OR=1.15, CI 0.97-1.37, P=.10) stages of change. CONCLUSIONS: In smokers in the contemplation stage of change and in former smokers, the original program produced higher smoking abstinence rates than the modified program. PMID- 15829475 TI - The emergence of national electronic health record architectures in the United States and Australia: models, costs, and questions. AB - Emerging electronic health record models present numerous challenges to health care systems, physicians, and regulators. This article provides explanation of some of the reasons driving the development of the electronic health record, describes two national electronic health record models (currently developing in the United States and Australia) and one distributed, personal model. The US and Australian models are contrasted in their different architectures ("pull" versus "push") and their different approaches to patient autonomy, privacy, and confidentiality. The article also discusses some of the professional, practical, and legal challenges that health care providers potentially face both during and after electronic health record implementation. PMID- 15829476 TI - Electronic health records should support clinical research. AB - One aspect of electronic care records which has received little attention is the potential benefit to clinical research. Electronic records could facilitate new interfaces between care and research environments, leading to great improvements in the scope and efficiency of research. Benefits range from systematically generating hypotheses for research to undertaking entire studies based only on electronic record data. Researchers and research managers must engage with electronic record initiatives to realize these benefits. Clinicians and patients must have confidence in the consent, confidentiality and security arrangements for the uses of secondary data. Provided that such initiatives establish adequate information governance arrangements, within a clear ethical framework, innovative clinical research should flourish. Major benefits to patient care could ensue given sufficient development of the care-research interface via electronic records. PMID- 15829477 TI - The Internet and clinical trials: background, online resources, examples and issues. AB - Both the Internet and clinical trials were significant developments in the latter half of the twentieth century: the Internet revolutionized global communications and the randomized controlled trial provided a means to conduct an unbiased comparison of two or more treatments. Large multicenter trials are often burdened with an extensive development time and considerable expense, as well as significant challenges in obtaining, backing up and analyzing large amounts of data. Alongside the increasing complexities of the modern clinical trial has grown the power of the Internet to improve communications, centralize and secure data as well as to distribute information. As more and more clinical trials are required to coordinate multiple trial processes in real time, centers are turning to the Internet for the tools to manage the components of a clinical trial, either in whole or in part, to produce lower costs and faster results. This paper reviews the historical development of the Internet and the randomized controlled trial, describes the Internet resources available that can be used in a clinical trial, reviews some examples of online trials and describes the advantages and disadvantages of using the Internet to conduct a clinical trial. We also extract the characteristics of the 5 largest clinical trials conducted using the Internet to date, which together enrolled over 26000 patients. PMID- 15829478 TI - Challenges of internet recruitment: a case study with disappointing results. AB - BACKGROUND: The Internet provides tremendous opportunities for innovative research, but few publications on the use of the Internet for recruiting study participants exist. This paper summarizes our experiences from 2 studies in which we attempted to recruit teenagers on the Internet for a questionnaire study to evaluate a smoking-cessation website. OBJECTIVE: To evaluate strategies of recruiting teenagers for the evaluation of a smoking-cessation website through the Internet. METHODS: In Study 1 (Defined Community Recruitment), we sent invitation emails to registered members of a youth health website, CyberIsle. A total of 3801 email addresses were randomly divided into 2 groups. In the first group, emails indicated that the first 30 respondents would receive a Can dollars 20 electronic gift certificate for use at an online bookstore if they would go to the Smoking Zine website and respond to a short survey. For the second group, the email also indicated that respondents would receive an additional Can dollars 10 gift certificate if they referred their friends to the study. Reminder emails were sent 10 days after the sending of the initial invitation email. In Study 2 (Open Recruitment), we posted invitation messages on Web discussion boards, Usenet forums, and one specialized recruitment website, and attempted a snowball recruiting strategy. When potential participants arrived at the study site, they were automatically randomized into either the higher incentives group (Can dollars 15 electronic gift certificate) or lower incentive group (Can dollars 5 gift certificate). RESULTS: In Study 1 (defined community recruitment), 2109 emails were successfully delivered. Only 5 subjects (0.24%), including 1 referred by a friend, passed the recruitment process and completed the questionnaire; a further 6 individuals visited the information page of the study but did not complete the study. In Study 2 (open recruitment), the number of users seeing the advertisement is unknown. A total of 35 users arrived at the website, of whom 14 participants were recruited (8 from the Can dollars 15 gift certificate group and 6 from the Can dollars 5 gift certificate group). Another 5 were recruited from the general Internet community (3 from discussion boards and 2 from the Research Volunteers website). The remaining 9 participants were recruited through friend referrals with the snowball strategy. CONCLUSIONS: Overall, the recruitment rate was disappointingly low. In our case, recruitment using Internet technologies including email, electronic discussion boards, Usenet forums, and websites did not prove to be an effective approach for soliciting young subjects to participate in our research. Possible reasons are discussed, including the participants' perspective. A major challenge is to differentiate trustable and legitimate messages from spam and fraudulent misinformation on the Internet. From the researchers' perspective, approaches are needed to engage larger samples, to verify participants' attributes, and to evaluate and adjust for potential biases associated with Internet recruitment. PMID- 15829479 TI - Usage and longitudinal effectiveness of a Web-based self-help cognitive behavioral therapy program for panic disorder. AB - BACKGROUND: Anxiety disorders are common problems that result in enormous suffering and economic costs. The efficacy of Web-based self-help approaches for anxiety disorders has been demonstrated in a number of controlled trials. However, there is little data regarding the patterns of use and effectiveness of freely available Web-based interventions outside the context of controlled trials. OBJECTIVE: To examine the use and longitudinal effectiveness of a freely available, 12-session, Web-based, cognitive behavioral therapy (CBT) program for panic disorder and agoraphobia. METHODS: Cumulative anonymous data were analyzed from 99695 users of the Panic Center. Usage statistics for the website were examined and a longitudinal survey of self-reported symptoms for people who registered for the CBT program was conducted. The primary outcome measures were self-reported panic-attack frequency and severity at the beginning of each session (sessions 2-12). RESULTS: Between September 1, 2002 and February 1, 2004, there were 484695 visits and 1148097 page views from 99695 users to the Panic Center. In that same time period, 1161 users registered for the CBT program. There was an extremely high attrition rate with only 12 (1.03%) out of 1161 of registered users completing the 12-week program. However, even for those who remained in the program less than 12 weeks we found statistically significant reductions (P<.002) in self-reported panic attack frequency and severity, comparing 2 weeks of data against data after 3, 6, or 8 weeks. For example, the 152 users completing only 3 sessions of the program reduced their average number of attacks per day from 1.03 (week 2) to 0.63 (week 3) (P<.001). CONCLUSIONS: Freely available Web-based self-help will likely be associated with high attrition. However, for the highly self-selected group who stayed in the program, significant improvements were observed. PMID- 15829480 TI - Pilot study of an Internet patient-physician communication tool for heart failure disease management. AB - BACKGROUND: Internet disease management has the promise of improving care in patients with heart failure but evidence supporting its use is limited. We have designed a Heart Failure Internet Communication Tool (HFICT), allowing patients to enter messages for clinicians, as well as their daily symptoms, weight, blood pressure and heart rate. Clinicians review the information on the same day and provide feedback. OBJECTIVE: This pilot study evaluated the feasibility and patients' acceptability of using the Internet to communicate with patients with symptomatic heart failure. METHODS: Patients with symptomatic heart failure were instructed how to use the Internet communication tool. The primary outcome measure was the proportion of patients who used the system regularly by entering information on average at least once per week for at least 3 months. Secondary outcomes measures included safety and maintainability of the tool. We also conducted a content analysis of a subset of the patient and clinician messages entered into the comments field. RESULTS: Between May 3, 1999 and November 1, 2002, 62 patients (mean age 48.7 years) were enrolled.. At 3 months 58 patients were alive and without a heart transplant. Of those, 26 patients (45%; 95% Confidence Interval, 0.33-0.58) continued using the system at 3 months. In 97% of all entries by participants weight was included; 68% of entries included blood pressure; and 71% of entries included heart rate. In 3386 entries out of all 5098 patient entries (66%), comments were entered. Functions that were not used included the tracking of diuretics, medications and treatment goals. The tool appeared to be safe and maintainable. Workload estimates for clinicians for entering a response to each patient's entry ranged from less than a minute to 5 minutes or longer for a detailed response. Patients sent 3386 comments to the Heart Function Clinic. Based on the content analysis of 100 patient entries, the following major categories of communication were identified: patient information; patient symptoms; patient questions regarding their condition; patient coordinating own care; social responses. The number of comments decreased over time for both patients and clinicians. CONCLUSION: While the majority of patients discontinued use, 45% of the patients used the system and continued to use it on average for 1.5 years. An Internet tool is a feasible method of communication in a substantial proportion of patients with heart failure. Further study is required to determine whether clinical outcomes, such as quality of life or frequency of hospitalization, are improved. PMID- 15829481 TI - What is eHealth (4): a scoping exercise to map the field. AB - BACKGROUND: Lack of consensus on the meaning of eHealth has led to uncertainty among academics, policymakers, providers and consumers. This project was commissioned in light of the rising profile of eHealth on the international policy agenda and the emerging UK National Programme for Information Technology (now called Connecting for Health) and related developments in the UK National Health Service. OBJECTIVES: To map the emergence and scope of eHealth as a topic and to identify its place within the wider health informatics field, as part of a larger review of research and expert analysis pertaining to current evidence, best practice and future trends. METHODS: Multiple databases of scientific abstracts were explored in a nonsystematic fashion to assess the presence of eHealth or conceptually related terms within their taxonomies, to identify journals in which articles explicitly referring to eHealth are contained and the topics covered, and to identify published definitions of the concept. The databases were Medline (PubMed), the Cumulative Index of Nursing and Allied Health Literature (CINAHL), the Science Citation Index (SCI), the Social Science Citation Index (SSCI), the Cochrane Database (including Dare, Central, NHS Economic Evaluation Database [NHS EED], Health Technology Assessment [HTA] database, NHS EED bibliographic) and ISTP (now known as ISI proceedings). We used the search query, "Ehealth OR e-health OR e*health". The timeframe searched was 1997-2003, although some analyses contain data emerging subsequent to this period. This was supplemented by iterative searches of Web-based sources, such as commercial and policy reports, research commissioning programmes and electronic news pages. Definitions extracted from both searches were thematically analyzed and compared in order to assess conceptual heterogeneity. RESULTS: The term eHealth only came into use in the year 2000, but has since become widely prevalent. The scope of the topic was not immediately discernable from that of the wider health informatics field, for which over 320000 publications are listed in Medline alone, and it is not explicitly represented within the existing Medical Subject Headings (MeSH) taxonomy. Applying eHealth as narrative search term to multiple databases yielded 387 relevant articles, distributed across 154 different journals, most commonly related to information technology and telemedicine, but extending to such areas as law. Most eHealth articles are represented on Medline. Definitions of eHealth vary with respect to the functions, stakeholders, contexts and theoretical issues targeted. Most encompass a broad range of medical informatics applications either specified (eg, decision support, consumer health information) or presented in more general terms (eg, to manage, arrange or deliver health care). However the majority emphasize the communicative functions of eHealth and specify the use of networked digital technologies, primarily the Internet, thus differentiating eHealth from the field of medical informatics. While some definitions explicitly target health professionals or patients, most encompass applications for all stakeholder groups. The nature of the scientific and broader literature pertaining to eHealth closely reflects these conceptualizations. CONCLUSIONS: We surmise that the field -- as it stands today -- may be characterized by the global definitions suggested by Eysenbach and Eng. PMID- 15829482 TI - Exosite-mediated substrate recognition of factor IX by factor XIa. The factor XIa heavy chain is required for initial recognition of factor IX. AB - Studies of the mechanisms of blood coagulation zymogen activation demonstrate that exosites (sites on the activating complex distinct from the protease active site) play key roles in macromolecular substrate recognition. We investigated the importance of exosite interactions in recognition of factor IX by the protease factor XIa. Factor XIa cleavage of the tripeptide substrate S2366 was inhibited by the active site inhibitors p-aminobenzamidine (Ki 28 +/- 2 microM) and aprotinin (Ki 1.13 +/- 0.07 microM) in a classical competitive manner, indicating that substrate and inhibitor binding to the active site was mutually exclusive. In contrast, inhibition of factor XIa cleavage of S2366 by factor IX (Ki 224 +/- 32 nM) was characterized by hyperbolic mixed-type inhibition, indicating that factor IX binds to free and S2366-bound factor XIa at exosites. Consistent with this premise, inhibition of factor XIa activation of factor IX by aprotinin (Ki 0.89 +/- 0.52 microM) was non-competitive, whereas inhibition by active site inhibited factor IXa beta was competitive (Ki 0.33 +/- 0.05 microM). S2366 cleavage by isolated factor XIa catalytic domain was competitively inhibited by p aminobenzamidine (Ki 38 +/- 14 microM) but was not inhibited by factor IX, consistent with loss of factor IX-binding exosites on the non-catalytic factor XI heavy chain. The results support a model in which factor IX binds initially to exosites on the factor XIa heavy chain, followed by interaction at the active site with subsequent bond cleavage, and support a growing body of evidence that exosite interactions are critical determinants of substrate affinity and specificity in blood coagulation reactions. PMID- 15829483 TI - A systematic mutagenesis study of Ile-282 in transmembrane segment M4 of the plasma membrane H+-ATPase. AB - Homology models of plasma membrane H(+)-ATPase (Bukrinsky, J. T., Buch-Pedersen, M. J., Larsen, S., and Palmgren, M. G. (2001) FEBS Lett. 494, 6-10) has pointed to residues in transmembrane segment M4 as being important for proton translocation by P-type proton pumps. To test this model, alanine-scanning mutagenesis was carried out through 12 residues in the M4 of the plant plasma membrane H(+)-ATPase AHA2. An I282A mutation showed apparent reduced H(+) affinity, and this residue was subsequently substituted with all other naturally occurring amino acids by saturation mutagenesis. The ability of mutant enzymes to substitute for the yeast proton pump PMA1 was found to correlate with the size of the side chain rather than its chemical nature. Thus, smaller side chains (Gly, Ala, and Ser) at this position resulted in lower H(+) affinity and lowered levels of H(+) transport in vivo, whereas substitution with side chains of similar and larger size resulted in only minor effects. Substitutions of Ile-282 had only minor effects on ATP affinity and sensitivity toward vanadate, ruling out an indirect effect through changes in the enzyme conformational equilibrium. These results are consistent with a model in which the backbone carbonyl oxygen of Ile 282 contributes directly to proton translocation. PMID- 15829484 TI - Stearoyl-CoA desaturase 1 deficiency increases CTP:choline cytidylyltransferase translocation into the membrane and enhances phosphatidylcholine synthesis in liver. AB - Stearoyl-CoA desaturase (SCD) is the rate-limiting enzyme in monounsaturated fatty acid synthesis. Previously, we showed that Scd1 deficiency reduces liver triglyceride accumulation and considerably decreases synthesis of very low density lipoprotein and its secretion in both lean and obese mice. In the present study, we found that Scd1 deficiency significantly modulates hepatic glycerophospholipid profile. The content of phosphatidylcholine (PC) was increased by 40% and the activities of CTP:choline cytidylyltransferase (CCT), the rate-limiting enzyme in de novo PC synthesis, and choline phosphotransferase were increased by 64 and 53%, respectively, in liver of Scd1-/- mice. In contrast, the protein level of phosphatidylethanolamine N-methyltransferase, an enzyme involved in PC synthesis via methylation of phosphatidylethanolamine, was decreased by 80% in the liver of Scd1-/- mice. Membrane translocation of CCT is required for its activation. Immunoblot analyses demonstrated that twice as much CCTalpha was associated with plasma membrane in livers of Scd1-/- compared with wild type mice, suggesting that Scd1 mutation leads to an increase in CCT membrane affinity. The incorporation of [(3)H]glycerol into PC was increased by 2.5-fold in Scd1-/- primary hepatocytes compared with those of wild type mice. Furthermore, mitochondrial glycerol-3-phosphate acyltransferase activity was reduced by 42% in liver of Scd1-/- mice; however, the activities of microsomal glycerol-3-phosphate acyltransferase, diacylglycerol acyltransferase, and ethanolamine phosphotransferase were not affected by Scd1 mutation. Our study revealed that SCD1 deficiency specifically increases CCT activity by promoting its translocation into membrane and enhances PC biosynthesis in liver. PMID- 15829485 TI - Activation of the lipid droplet controls the rate of lipolysis of triglycerides in the insect fat body. AB - The hydrolysis of triglyceride (TG) stored in the lipid droplets of the insect fat body is under hormonal regulation by the adipokinetic hormone (AKH), which triggers a rapid activation cAMP-dependent kinase cascade (protein kinase A (PKA)). The role of phosphorylation on two components of the lipolytic process, the TG-lipase and the lipid droplet, was investigated in fat body adipocytes. The activity of purified TG-lipase determined using in vivo TG-radiolabeled lipid droplets was unaffected by the phosphorylation of the lipase. However, the activity of purified lipase was 2.4-fold higher against lipid droplets isolated from hormone-stimulated fat bodies than against lipid droplets isolated from unstimulated tissue. In vivo stimulation of lipolysis promotes a rapid phosphorylation of a lipid droplet protein with an apparent mass of 42-44 kDa. This protein was identified as "Lipid Storage Droplet Protein 1" (Lsdp1). In vivo phosphorylation of this protein reached a peak approximately 10 min after the injection of AKH. Supporting a role of Lsdp1 in lipolysis, maximum TG-lipase activity was also observed with lipid droplets isolated 10 min after hormonal stimulation. The activation of lipolysis was reconstituted in vitro using purified insect PKA and TG-lipase and lipid droplets. In vitro phosphorylation of lipid droplets catalyzed by PKA enhanced the phosphorylation of Lsdp1 and the lipolytic rate of the lipase, demonstrating a prominent role PKA and protein phosphorylation on the activation of the lipid droplets. AKH-induced changes in the properties of the substrate do not promote a tight association of the lipase with the lipid droplets. It is concluded that the lipolysis in fat body adipocytes is controlled by the activation of the lipid droplet. This activation is achieved by PKA-mediated phosphorylation of the lipid droplet. Lsdp1 is the main target of PKA, suggesting that this protein is a major player in the activation of lipolysis in insects. PMID- 15829486 TI - Mental disorders in prison populations aged 15-21: national register study of two cohorts in Finland. PMID- 15829487 TI - Metabolic syndrome criteria: ready for clinical prime time or work in progress? PMID- 15829488 TI - Management of asymptomatic aortic stenosis: how helpful is exercise testing? PMID- 15829489 TI - Bridging the practice chasm. PMID- 15829490 TI - Simple enumerations of peripheral blood natural killer (CD56+ NK) cells, B cells and T cells have no predictive value in IVF treatment outcome. AB - BACKGROUND: To evaluate the association between the absolute counts of the peripheral natural killer (NK) cells (including total CD56(+) NK cells, CD56(dim) NK cells and CD56(bright) NK cells), B cells and T cells on the implantation rate and miscarriage rate after IVF treatment. METHODS: This was a prospective observation study. A total of 138 patients who underwent IVF treatment from December 2002 to July 2003 were recruited to the study. Blood samples were obtained on the day of vaginal oocyte retrieval prior to the procedure. The absolute counts of lymphocytes, NK cells, B cells and T cells were identified by flow cytometry. These absolute counts and their relationships to IVF treatment outcome and miscarriage rate were analysed. RESULTS: There were no significant differences with regard the mean values of absolute lymphocyte count, T cell count, B cell count and NK cell count (including total CD56(+) NK, CD56(dim) NK and CD56(bright) NK cells) between the pregnant and non-pregnant groups and also between the ongoing pregnancy and miscarriage groups. The cause of infertility, duration of infertility, basal FSH levels, number of previous failed IVF treatments, number of previous miscarriages and stimulation characteristics were not significantly different between the pregnant and non-pregnant groups. Previous studies have suggested that women with a history of recurrent miscarriage and those with infertility accompanied by recurrent failed IVF treatments are associated with a peripheral blood NK cell percentage >12%, therefore further analysis of peripheral CD56(+) NK cell levels <12% (group A) and >12% (group B) was performed. There was no significant difference in implantation rate (group A: 17.0%; group B: 23.2%), pregnancy rate (group A: 36.6%; group B: 47.7%) or miscarriage rate (group A: 23.3%; group B: 28.6%). CONCLUSION: There were no significant differences between simple enumerations of peripheral blood NK cells (including total CD56(+) NK, CD56(dim) NK and CD56(bright) NK cells), B cells and T cells with IVF treatment outcome and pregnancy outcome. Women who had a peripheral NK cell level >12% did not have higher number of previous pregnancy losses. Importantly their pregnancy rate was not reduced and their miscarriages were not increased compared to women who had a peripheral NK cells level <12%. PMID- 15829491 TI - Safety of sentinel node biopsy in pregnant patients. PMID- 15829493 TI - Therapeutic vaccination in patients with gastrointestinal malignancies. A review of immunological and clinical results. AB - Gastrointestinal (GI) malignancies are the most common types of human cancers. Despite the introduction of new cytotoxic drugs, a large proportion remains incurable. There is a great need to develop new complementary therapeutic modalities. Strategies exploiting targeted therapies are expanding. The focus of the present article is to review active specific immunotherapy (vaccination) in patients with GI malignancies. The review comprises a description of the immunogenicity of GI malignancies, various types of tumour antigens and mechanisms of action of cancer vaccines. Tumour escape from immune surveillance, vaccine strategies and adjuvants are also described. Clinical and immunological endpoints of cancer immunotherapy are outlined. Results of therapeutic vaccine trials published mainly during the last 5 years in PubMed enrolling a minimum of six patients with GI malignancies are included. Studies presented at the two last annual meetings of the American Society of Clinical Oncology are also covered. More than 2000 patients have been vaccinated with tumour antigens (self antigens). The procedure is safe and no autoimmune disorders have been observed after >4 years follow-up in a substantial number of patients. Humoral and cellular tumour antigen-specific immune responses were induced. A correlation between immune responses and prolonged overall survival was seen in several studies. The most encouraging results were noted in randomised controlled phase II/III trials including over 1300 colorectal carcinoma patients with minimal residual disease. A statistically significantly improved disease-free or overall survival was shown either in all vaccinated or in sub-groups of patients. Promising results were also reported in pancreatic and hepatocellular carcinoma. If the results of the randomised controlled trials hold true, active specific immunotherapy may provide a new promising targeted therapeutic approach in GI malignancies with minimal toxicity. Further enlarged randomised controlled studies are warranted to confirm the results, particularly in colon carcinoma with minimal residual disease. PMID- 15829494 TI - Temozolomide and cisplatin versus temozolomide in patients with advanced melanoma: a randomized phase II study of the Hellenic Cooperative Oncology Group. AB - PURPOSE: Temozolomide (TMZ) is an oral alkylating agent that produces methyl adducts at the 0.6 position of guanine. The methyl adducts are removed by the DNA repair enzyme AGAT. As demonstrated by in vitro studies, cisplatin (CDDP) is able to down-regulate the AGAT activity, suggesting that CDDP could enhance the antitumor activity of TMZ. We designed a randomized phase II study to evaluate and compare the activity and safety profile of the combination versus single agent TMZ in patients with advanced melanoma. PATIENTS AND METHODS: From January 2000 to April 2002, 132 patients were enrolled on the study. Patient and tumor characteristics were well balanced between the two arms. Patients with cerebral metastases were included. Patients received TMZ 200 mg/m(2)/day orally for five consecutive days every 4 weeks or TMZ + CDDP 200 mg/m(2) daily on days 1-5 and 75 mg/m(2) of CDDP on day 1. RESULTS: Tumor responses (complete and partial responses) were seen in 16 patients (26%) in arm A and 19 patients (29%) in arm B. The median time to progression (TTP) was 3.8 months in arm A and 5.8 months in arm B. The median overall survival (OS) was 11.5 months in arm A and 12 months in arm B. The difference between treatment arms regarding objective response rates, TTP and OS were not statistically significant. Toxicity was comparable between the two arms for anemia, leukopenia, neutropenia, thrombocytopenia, fatigue, constipation and arthralgias/myalgias. There was significantly more grade 3 and 4 emesis in the combination arm. CONCLUSIONS: No clear benefit in terms of response rates, median TTP or OS was shown with the combination of TMZ + CDDP. Additionally, the combination was associated with higher incidence of grade 3 and 4 emesis. PMID- 15829495 TI - Analysis of the dinucleotide repeat polymorphism in the epidermal growth factor receptor (EGFR) gene in head and neck cancer patients. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) overexpression is associated with poor prognosis in head and neck cancer. The first intron of EGFR gene is polymorphic (9-23 CA repeats) and transcription declines when the number of repeats increases. PATIENTS AND METHODS: EGFR polymorphism (fluorescent genotyping) and expression (ligand-binding assay) were analyzed in tumors and normal tissues from 112 patients (100 men, 12 women; mean age 60 years). RESULTS: The number of CA repeats varied from 15 to 22. Allelic distribution was trimodal (predominance of 16, 20 and 18 CA repeats). EGFR concentrations were significantly higher (P=0.02) in homozygous tumors as compared with heterozygous. Considering homozygous tumors, or classifying genotypes as short/long/intermediary (two alleles <17 versus two alleles > or =17 versus others), no relationship was observed between tumoral EGFR genotype and expression. In the 76 tumors exhibiting at least one 16-CA allele, the length of the remaining allele was inversely correlated to EGFR expression (P=0.047). Tumoral EGFR expression, performance status (WHO criteria) and node involvement were independent predictors of specific survival (P <0.01). Tumoral or normal tissue EGFR genotype did not influence survival. CONCLUSIONS: Intron 1 EGFR polymorphism may be implicated in the regulation of EGFR expression in head and neck tumors. PMID- 15829496 TI - 'Stop it, it's bad for you and me': experiences of and views on passive smoking among primary-school children in Liverpool. AB - This article looks at how children between the ages of 4 and 8 years report they feel when they are exposed to passive smoking and how they react in these situations. Data were collected annually from a cohort of 250 primary school children, which was tracked from their Reception Classes to Year 3 across six Liverpool schools. Quantitative and qualitative methods were employed--including a survey, the Draw and Write investigative technique and semi-structured interviews. Findings showed that children had some understanding of the health problems that passive smoking posed to both themselves and the smoker. Between the ages of 4 and 7 the majority of children held negative feelings about being exposed to smoke, but at the age of 8 there is a clear decline in negative expressions. Most of the children were prepared verbally to confront a smoker, usually a parent, in order to get them to stop, but rarely took direct action and left the room themselves. Results suggest that children would be receptive to information on the dangers of smoking during the early years of primary school, while the dialogue between children and their parents suggests that the latter have a key role to play in strategies to tackle passive smoking in the home. PMID- 15829497 TI - Phytoestrogen exposure elevates PTEN levels. AB - Epidemiological data suggest that consumption of phytoestrogens can be protective against the development of breast cancer. It may be logical to postulate that phytoestrogens may regulate proteins that control cellular division, such as the tumor suppressor PTEN. Germline, and more significantly, somatic PTEN mutations have been observed in a broad range of human cancers, especially those of the breast. Active PTEN results in decreased phosphorylation of Akt and MAPK, the up regulation of p27 and down-regulation of cyclin D1 protein levels resulting in decreased proliferation and an increase in apoptosis. We hypothesized that phytoestrogen exposure regulates PTEN protein expression in the breast cancer cell line, MCF-7. When MCF-7 cells were stimulated with resveratrol, quercetin or genistein, there was an increase in PTEN protein levels. Concomitantly, phytoestrogen stimulation resulted in decreased Akt phosphorylation and an increase in p27 protein levels, indicating active PTEN lipid phosphatase activity. In contrast, we found that MAPK phosphorylation and cyclin D1 levels, which are regulated by PTEN's protein phosphatase activity, were not altered. Using semi-quantitative RT-PCR, we found that mRNA levels were slightly increased in cells stimulated by phytoestrogens, suggesting that the mechanism for increased PTEN protein expression is dependent upon transcription. Concurrently, our data provide evidence that a mechanism for phytoestrogens' protective nature is partially through increased PTEN expression. More importantly, it provides a novel target for the regulation of PTEN expression and suggests that dietary changes may be adjunctive to traditional preventive and therapeutic strategies against breast cancer. PMID- 15829498 TI - Toll-like receptor 4 variant D299G is associated with susceptibility to age related macular degeneration. AB - Age-related macular degeneration (AMD) is a genetically heterogeneous disease that leads to progressive and irreversible vision loss among the elderly. Inflammation, oxidative damage, cholesterol metabolism and/or impaired function of retinal pigment epithelium (RPE) have been implicated in AMD pathogenesis. We examined toll-like receptor 4 (TLR4) as a candidate gene for AMD susceptibility because: (i) the TLR4 gene is located on chromosome 9q32-33, a region exhibiting evidence of linkage to AMD in three independent reports; (ii) the TLR4-D299G variant is associated with reduced risk of atherosclerosis, a chronic inflammatory disease with subendothelial accumulation; (iii) the TLR4 is not only a key mediator of proinflammatory signaling pathways but also linked to regulation of cholesterol efflux and (iv) the TLR4 participates in phagocytosis of photoreceptor outer segments by the RPE. We examined D299G and T399I variants of TLR4 in a sample of 667 unrelated AMD patients and 439 unrelated controls, all of Caucasian ancestry. Multiple logistic regression demonstrated an increased risk of AMD in carriers of the G allele at TLR4 residue 299 (odds ratio=2.65, P=0.025), but lack of an independent effect by T399I variant. TLR4-D299G showed an additive effect on AMD risk (odds ratio=4.13, P=0.002) with allelic variants of apolipoprotein E (APOE) and ATP-binding cassette transporter-1 (ABCA1), two genes involved in cholesterol efflux. Interestingly, the effect of TLR4, APOE and ABCA1 variants on AMD susceptibility was opposite to that of association with atherosclerosis risk. Our data provide evidence of a link between multiple diverse mechanisms underlying AMD pathogenesis. PMID- 15829499 TI - The Charcot-Marie-Tooth type 2A gene product, Mfn2, up-regulates fuel oxidation through expression of OXPHOS system. AB - Mitofusin-2 (Mfn2) is a mitochondrial membrane protein that participates in mitochondrial fusion in mammalian cells and mutations in the Mfn2 gene cause Charcot-Marie-Tooth neuropathy type 2A. Here, we show that Mfn2 loss-of-function inhibits pyruvate, glucose and fatty acid oxidation and reduces mitochondrial membrane potential, whereas Mfn2 gain-of-function increases glucose oxidation and mitochondrial membrane potential. As to the mechanisms involved, we have found that Mfn2 loss-of-function represses nuclear-encoded subunits of OXPHOS complexes I, II, III and V, whereas Mfn2 overexpression induced the subunits of complexes I, IV and V. Obesity-induced Mfn2 deficiency in rat skeletal muscle was also associated with a decrease in the subunits of complexes I, II, III and V. In addition, the effect of Mfn2 overexpression on mitochondrial metabolism was mimicked by a truncated Mfn2 mutant that is inactive as a mitochondrial fusion protein. Our results indicate that Mfn2 triggers mitochondrial energization, at least in part, by regulating OXPHOS expression through signals that are independent of its role as a mitochondrial fusion protein. PMID- 15829500 TI - Identification and characterization of a novel gene Saf transcribed from the opposite strand of Fas. AB - Apoptosis is a morphologically distinct form of cell death involved in many physiological and pathological processes. The regulation of Fas/Apo-1 involved in membrane-mediated apoptosis has also been known to play crucial roles in many systems. More and more naturally occurring antisense RNAs are now known to regulate, at least in part, a growing number of eukaryotic genes. In this report, we describe the findings of a novel RNA transcribed from the opposite strand of the intron 1 of the human Fas gene. Using orientation-specific RT-PCR and northern blot analysis, we show that this transcript is 1.5 kb in length and was expressed in several human tissues and cell lines. This transcript was cloned by 5'- and 3'-RACE (rapid amplification of cDNA ends) and the transcription start site was determined by primer extension. This novel gene was named Saf. To assess the functions of Saf, Jurkat cells transfected with human Saf or control vector was prepared. The stable Saf-transfectant was highly resistant to Fas-mediated but not to TNF-alpha-mediated apoptosis. Although the overall mRNA expression level of Fas was not affected, expression of some novel forms of Fas transcripts was increased in Saf-transfectant, especially the inhibitory soluble forms. These findings collectively suggest that Saf might protect T lymphocytes from Fas mediated apoptosis by blocking the binding of FasL or its agonistic Fas antibody. Saf might regulate the expression of Fas alternative splice forms through pre mRNA processing. PMID- 15829501 TI - Lowe syndrome protein Ocrl1 is translocated to membrane ruffles upon Rac GTPase activation: a new perspective on Lowe syndrome pathophysiology. AB - Oculocerebrorenal Lowe syndrome is a rare X-linked disorder characterized by bilateral cataract, mental retardation and renal Fanconi syndrome. The Lowe syndrome protein Ocrl1 is a PIP2 5-phosphatase, primarily localized to the trans Golgi network (TGN), which 'loss of function' mutations result in PIP2 accumulation in patient's cells. Although PIP2 is involved in many cell functions including signalling, vesicle trafficking and actin polymerization, it has been difficult so far to decipher molecular/cellular mechanisms responsible for Lowe syndrome phenotype. We have recently shown that, through its C-terminal RhoGAP domain, Ocrl1 forms a stable complex with Rac GTPase within the cell. In line with this finding, we report here that upon epidermal growth factor induced Rac activation in COS-7 cells, a fraction of Ocrl1 translocates from TGN to plasma membrane and concentrates in membrane ruffles. In order to investigate the functionality of Ocrl1 in plasma membrane, we have analysed PIP2 distribution in human dermal fibroblasts (HDFs) from Lowe patients versus control HDFs. As revealed by both immunodetection and green fluorescent protein-PH binding, PIP2 was found strikingly to accumulate in PDGF induced ruffles in Lowe HDFs when compared with control. This suggests that Ocrl1 is active as a PIP2 5-phosphatase in Rac induced membrane ruffles. Cellular properties such as cell migration and establishment of cell-cell contacts, which depend on ruffling and lamellipodia formation, should be further investigated to understand the pathophysiology of Lowe syndrome. PMID- 15829502 TI - A role for fibroblast growth factor receptor-2 in the altered osteoblast phenotype induced by Twist haploinsufficiency in the Saethre-Chotzen syndrome. AB - Genetic mutations of Twist, a bHLH transcription factor, induce premature fusion of cranial sutures (craniosynostosis) in the Saethre-Chotzen syndrome (SCS). The mechanisms by which Twist haploinsufficiency may alter osteoblast differentiation are poorly understood. In this study, we investigated the role of fibroblast growth factor receptor-2 (Fgfr2) in the abnormal osteoblast differentiation in SCS. Cranial osteoblasts from an SCS patient with a Y103X mutation inducing deletion of the Twist bHLH domain showed decreased Fgfr2 mRNA levels associated with decreased expression of Runx2, bone sialoprotein (BSP) and osteocalcin (OC), markers of differentiated osteoblasts, compared with wild-type osteoblasts. Transfection with Twist or Runx2 expression vectors, but not with Runx2 mutant which impairs DNA binding, restored Fgfr2, Runx2, BSP and OC expression in Twist mutant osteoblasts. EMSA analysis of mutant osteoblast nuclear extracts showed reduced Runx2 binding to a target OSE2 site in the Fgfr2 promoter. ChIP analyses showed that both Twist and Runx2 in mutant osteoblast nuclear extracts bind to a specific region in the Fgfr2 promoter. Significantly, forced expression of Fgfr2 restored Runx2 and osteoblast marker genes, whereas a dominant-negative Fgfr2 further decreased Runx2 and downstream genes in Twist mutant osteoblasts, indicating that alteration of Fgfr2 results in downregulation of osteoblast genes in Twist mutant osteoblasts. We conclude that Twist haploinsufficiency downregulates Fgfr2 mRNA expression, which in turn reduces Runx2 and downstream osteoblast-specific genes in human calvarial osteoblasts. This provides genetic and biochemical evidence for a role of Fgfr2 in the altered osteoblast phenotype induced by Twist haploinsufficiency in the SCS. PMID- 15829503 TI - SINE exonic insertion in the PTPLA gene leads to multiple splicing defects and segregates with the autosomal recessive centronuclear myopathy in dogs. AB - Human centronuclear and myotubular myopathies belong to a genetically heterogeneous nosological group with clinical variability ranging from fatal disorder to mild weakness. The severe X-linked form is attributed to more than 200 different mutations in the myotubularin encoding gene (MTM1). In contrast, there are no reports regarding the molecular etiology or linkage studies on the autosomal forms of the disease. Labrador retrievers affected by spontaneous centronuclear myopathy (cnm) have clinical and histological features of the human disorder and represent the first model of recessive autosomal centronuclear myopathy. We previously mapped the cnm locus to the centromeric region of canine chromosome 2. No gene of the MTM1 family maps to the human homologous chromosomal region. Described herein is a disease-associated insertion within PTPLA exon 2, found in both alleles of all affected Labradors and in a single allele in obligate carriers. The inserted tRNA-derived short interspersed repeat element (SINE) has a striking effect on the maturation of PTPLA mRNA, whereby it can be spliced out, partially exonized or involved in multiple exon-skipping. As a result, the amount of wild-type transcripts falls to 1% in affected muscles. This example therefore recapitulates cumulative SINE-associated transcriptional defects that have been previously described as exclusive consequences of independent mutations. Although the function of PTPLA in metazoa remains unknown, the characterization of a hypomorphic mutation in Labradors with centronuclear myopathy provides new clues about the molecular complexity of skeletal myofiber homeostasis. These results also suggest that impaired PTPLA signaling might be implicated in human myopathies. PMID- 15829504 TI - SLC18A2 promoter haplotypes and identification of a novel protective factor against alcoholism. AB - The vesicular monoamine transporter 2 (VMAT2, SLC18A2) takes up cytosolic monoamines into intracellular secretory vesicles, preventing their neurotoxicity in the cytosol and discharging them into extracellular space by exocytosis. It has been shown that one-copy deletion of the VMAT2 gene increases locomotion activity significantly in response to drug treatments and dopamine neuron death rate in response to neurotoxin treatments in knockout mice. Little is known about promoter polymorphisms and their influence on SLC18A2 promoter activity. We have re-sequenced a 17.4 kb DNA in the SLC18A2 promoter region for Caucasians and revealed 47 polymorphisms that confer 13 haplotypes. One of the haplotypes reaches a frequency as high as 65%, likely due to positive selection. In vitro analysis showed a 20% difference in promoter activity between two frequent haplotypes and identified some of the polymorphisms that influence promoter activity. Four haplotype-defining single nucleotide polymorphisms (hdSNPs) can define the frequent haplotypes and by genotyping these hdSNPs, we find that haplotypes with -14234G and -2504C of SLC18A2 promoter region represent a protective factor against alcoholism (P = 0.0038 by Fisher's exact tests). Therefore, SLC18A2 promoter haplotypes defined here create a foundation for transcriptional characterization of individuality and for association study on monoamine-related human diseases. PMID- 15829505 TI - Loss of wild-type huntingtin influences motor dysfunction and survival in the YAC128 mouse model of Huntington disease. AB - Huntington disease (HD) is an adult-onset neurodegenerative disease caused by a toxic gain of function in the huntingtin (htt) protein. The contribution of wild type htt function to the pathogenesis of HD is currently uncertain. To assess the role of wild-type htt in HD, we generated YAC128 mice that do not express wild type htt (YAC128-/-) but express the same amount of mutant htt as normal YAC128 mice (YAC128+/+). YAC128-/- mice perform worse than YAC128+/+ mice in the rotarod test of motor coordination (P = 0.001) and are hypoactive compared with YAC128+/+ mice at 2 months (P = 0.003). Striatal neuropathology was not clearly worse in YAC128-/- mice compared with YAC128+/+ mice. There was no significant effect of decreased wild-type htt on striatal volume, neuronal counts or DARPP-32 expression but a modest worsening of striatal neuronal atrophy was evident (6%, P = 0.03). The testis of YAC128+/+ mice showed atrophy and degeneration, which was markedly worsened in the absence of wild-type htt (P = 0.001). YAC128+/+ mice also showed a male specific deficit in survival compared with WT mice which was exacerbated by the loss of wild-type htt (12-month-male survival, P < 0.001). Overall, we demonstrate that the loss of wild-type htt influences motor dysfunction, hyperkinesia, testicular degeneration and impaired lifespan in YAC128 mice. The mild effect of wild-type htt on striatal phenotypes in YAC128 mice suggests that the characteristic striatal neuropathology in HD is caused primarily by the toxicity of mutant htt and that replacement of wild-type htt will not be an adequate treatment for HD. PMID- 15829506 TI - Fine-tuning in Ca2+ homeostasis underlies progression of cardiomyopathy in myocytes derived from genetically modified embryonic stem cells. AB - Mutations of genes encoding contractile proteins are responsible for familial hypertrophic cardiomyopathies. Understanding the process of differentiation of cardiomyocytes carrying a mutated protein is a crucial step towards potential treatments of inherited cardiac disorders. Embryonic Stem (ES) cells which faithfully recapitulate in vitro the process of cardiac cell differentiation can be genetically modified to incorporate a mutation mimicking a cardiomyopathy. ES cell lines engineered to express a wild-type (MLC2vGFP) or a mutated form (R58QMLC2vGFP) of ventricular myosin light chain 2 (MLC2v) fused to GFP were differentiated into cardiomyocytes within embryoid bodies (EBs). Visualization of GFP combined with sarcomeric actinin immunofluorescence of EBs revealed that mutated MLC2v dramatically prevented myofibrillogenesis. Cardiomyocytes expressing wild-type MLC2v featured spontaneous Ca(2+) spiking, but not those harboring the mutation. Expression of cardiac transcription factors Mef2c, GATAs, myocardin and Nkx2.5 was not affected by cell expression of mutated MLC2v. A dramatic decrease in expression of mRNAs encoding alpha-actin, MLC2a and MLC2v was observed in R58QMLC2vGFP EBs. This event was attributed to a failure of Mef2c to translocate into the nucleus, a Ca(2+)-dependent process. Expression in mutated cells of a constitutively active Ca(2+)- and calmodulin-dependent kinase II or treating EBs with ionomycin fully restored translocation of Mef2c into the nucleus and expression of mRNAs encoding sarcomeric proteins partially rescued contractile activity of EBs. Alteration of Ca(2+) homeostasis in mutated cardioblasts affects the transcriptional program of cardiac cell differentiation leading to a defect in myofibrillogenesis, and, in turn, in contractility. Genetically modified ES cells provide a unique cell model to determine abnormalities in Ca(2+) homeostasis underlying progression of human cardiomyopathies. PMID- 15829507 TI - Intra-nuclear trafficking of the BLM helicase to DNA damage-induced foci is regulated by SUMO modification. AB - The Bloom syndrome gene, BLM, encodes a RecQ DNA helicase that when absent from the cell results in genomic instability and cancer predisposition. We show here that BLM is a substrate for small ubiquitin-like modifier (SUMO) modification, with lysines at K317, K331, K334 and K347 being preferred sites of modification. Unlike normal BLM, a double mutant BLM protein with lysine to arginine substitutions at residues 317 and 331 was not modified by SUMO, and it failed to localize efficiently to the PML nuclear bodies. Rather, double mutant BLM protein induced the formation of DNA damage-induced foci (DDI) that contained BRCA1 protein and phosphorylated histone H2AX. Double mutant BLM only partially complemented the genomic instability phenotypes of Bloom syndrome cells as assessed by sister-chromatid exchange and micronuclei formation assays. These results constitute evidence that BLM is a DNA damage sensor that signals the formation of DDI, and they establish SUMO modification as a negative regulator of BLM's signaling function. PMID- 15829508 TI - Effects of aluminum on light energy utilization and photoprotective systems in citrus leaves. AB - BACKGROUND AND AIMS: Under high photon flux, excitation energy may be in excess in aluminum (Al)-treated leaves, which use a smaller fraction of the absorbed light in electron transport due to decreased CO2 assimilation compared with normal leaves. The objectives of this study were to test the hypothesis that the antioxidant systems are up-regulated in Al-treated citrus leaves and correlate with protection from photoxidative damage, and to test whether xanthophyll cycle dependent thermal energy dissipation is involved in dissipating excess excitation energy. * METHODS: 'Cleopatra' tangerine seedlings were fertilized and irrigated daily for 8 weeks with quarter-strength Hoagland's nutrient solution containing Al at a concentration of 0 or 2 mM from Al2(SO4)3.18H2O. Thereafter, leaf absorptance, chlorophyll (Chl) fluorescence, Al, pigments, antioxidant enzymes and metabolites were measured on fully expanded leaves. * KEY RESULTS: Compared with control leaves, energy was in excess in Al-treated leaves, which had smaller thermal energy dissipation, indicated by non-photochemical quenching (NPQ). In contrast, conversion of violaxanthin (V) to antheraxanthin (A) and zeaxanthin (Z) at midday increased in both treatments, but especially in Al-treated leaves, although A + Z accounted for less 40 % of the total xanthophyll cycle pool in them. Activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and catalase (CAT), and concentrations of ascorbate (AsA), dehydroascorbate (DASA), reduced glutathione (GSH) and oxidized glutathione (GSSG) were higher in Al-treated than in control leaves. * CONCLUSIONS: These results corroborate the hypothesis that, compared with control leaves, antioxidant systems are up-regulated in Al-treated citrus leaves and protect from photoxidative damage, whereas thermal energy dissipation was decreased. Thus, antioxidant systems are more important than thermal energy dissipation in dissipating excess excitation energy in Al-treated citrus leaves. PMID- 15829510 TI - Photosynthesis in relation to reproductive success of Cypripedium flavum. AB - BACKGROUND AND AIMS: Cypripedium flavum is a rare, endemic alpine slipper orchid of China, which is under threat from excessive collection and habitat changes. Conservation and re-introduction of C. flavum is restricted by lack of knowledge of the plant's photosynthesis and how that affects reproductive success. The hypothesis is tested that reproductive success is determined by photosynthetic production. * METHODS: To understand the photosynthetic characteristics and adaptation of C. flavum to alpine environments, and the relation to reproductive success, measurements were made at four field sites with varying degrees of forest cover in the Hengduan Mountains, south-west China. * KEY RESULTS: Both photosynthetic capacity and reproductive traits of C. flavum are affected by light availability. Photosynthetic rate (A) is greatest around noon, following the pattern of photosynthetically active radiation (PAR) at all sites. Cypripedium flavum has highest daily mean photosynthetic rate (A(daily)) and light-saturated photosynthetic rate (A(max)) under a half to a third of full sunlight. High radiation decreased A. However, the optimum temperature for photosynthesis was similar (18-20 degrees C) at all sites. * CONCLUSIONS: The quotient of daily mean photosynthetic rate to light saturated photosynthesis (A(daily)/A(max)) is positively correlated with the ramet number m(-2) and percentage of fruiting of C. flavum. The A(daily)/A(max) ratio is a useful proxy for evaluating reproductive success of C. flavum. PMID- 15829509 TI - Ancestry of American polyploid Hordeum species with the I genome inferred from 5S and 18S-25S rDNA. AB - BACKGROUND AND AIMS: The genus Hordeum exists at three ploidy levels (2x, 4x and 6x) and presents excellent material for investigating the patterns of polyploid evolution in plants. Here the aim was to clarify the ancestry of American polyploid species with the I genome. * METHODS: Chromosomal locations of 5S and 18S-25S ribosomal RNA genes were determined by fluorescence in situ hybridization (FISH). In both polyploid and diploid species, variation in 18S-25S rDNA repeated sequences was analysed by the RFLP technique. * KEY RESULTS: Six American tetraploid species were divided into two types that differed in the number of rDNA sites and RFLP profiles. Four hexaploid species were similar in number and location of both types of rDNA sites, but the RFLP profiles of 18S-25S rDNA revealed one species, H. arizonicum, with a different ancestry. * CONCLUSIONS: Five American perennial tetraploid species appear to be alloploids having the genomes of an Asian diploid H. roshevitzii and an American diploid species. The North American annual tetraploid H. depressum is probably a segmental alloploid combining the two closely related genomes of American diploid species. A hexaploid species, H. arizonicum, involves a diploid species, H. pusillum, in its ancestry; both species share the annual growth habit and are distributed in North America. Polymorphisms of rDNA sites detected by FISH and RFLP analyses provide useful information to infer the phylogenetic relationships of I-genome Hordeum species because of their highly conserved nature during polyploid evolution. PMID- 15829511 TI - Monitoring cytosolic calcium in the dinoflagellate Crypthecodinium cohnii with calcium orange-AM. AB - Calcium plays several important roles in the signal transduction pathways of dinoflagellates. We describe here the development of calcium orange-AM as an intracellular calcium reporter for the heterotrophic dinoflagellate Crypthecodinium cohnii. We demonstrated with confocal microscopy that by restricting the incubation period to 30-45 min, no compartmentalization of the dye occurs in the mitochondria or endoplasmic reticulum. The dye fluorescence responded well to the effects of calcium ionophores and calcium chelators. By calibrating the dye with known calcium concentrations, we determined the intracellular calcium concentration of C. cohnii to be 158 +/- 56 nM, which rose to about 550 nM upon mechanical stimulation. PMID- 15829512 TI - Nitric oxide is involved in methyl jasmonate-induced defense responses and secondary metabolism activities of Taxus cells. AB - Methyl jasmonate (MeJA), a methyl ester of jasmonic acid (JA), is a well established signal molecule in plant defense responses and an effective inducer of secondary metabolite accumulation in plant cell cultures such as the valuable anticancer diterpenoid taxol (paclitaxel) in Taxus spp. This work examines the involvement of nitric oxide (NO) in MeJA-induced plant defense responses and secondary metabolism in Taxus chinensis cell cultures. Exogenously supplied MeJA at 100 microM induced rapid production of NO in the Taxus cell cultures, reaching a maximum within 6 h of MeJA supply. Several other responses occurred concomitantly, including the production of hydrogen peroxide (H2O2), and the increases in intracellular malondialdehyde (MDA) content, lipoxygenase (LOX) and phenylalanine ammonium-lyase (PAL) activities. The MeJA-induced H2O2 production was suppressed by an NO donor, sodium nitroprusside (SNP), but enhanced by NO inhibitors, N (omega)-nitro-L-arginine (L-NNA) and 2-phenyl-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (PTIO). In contrast, the MeJA-induced MDA, LOX and PAL were all enhanced by the NO donor but suppressed by the NO inhibitors. The NO inhibitors also suppressed MeJA-induced taxol accumulation. These results are suggestive of a role for NO as a signal element for activating the MeJA-induced defense responses and secondary metabolism activities of plant cells. PMID- 15829513 TI - A novel receptor kinase involved in jasmonate-mediated wound and phytochrome signaling in maize coleoptiles. AB - We identified a gene of maize (Zea mays L.) that is transcriptionally activated in decapitated coleoptiles. The amino acid sequence deduced from its full-length cDNA indicated that the identified gene encodes a novel leucine-rich-repeat receptor-like kinase. The gene is named WOUND-RESPONSIVE AND PHYTOCHROME REGULATED KINASE1 (WPK1) based on the findings of this study. Database searches revealed two and three homologs of WPK1 for Arabidopsis thaliana and rice, respectively. These homologs occurred along with WPK1 on a phylogenetic branch separated from all reported receptor kinases. We uncovered that the level of WPK1 transcripts is up-regulated rapidly and transiently in response to wounding and red light. The response to red light was reversible by far-red light, indicating that it is mediated by phytochrome. Applied jasmonic acid activated the expression of WPK1, while ethylene, salicylic acid and abscisic acid had no such effect. These results strongly suggested that WPK1 is a component of the jasmonate-mediated signaling that participates in both wound-induced defensive and phytochrome-mediated photomorphogenetic responses. Furthermore, it was found that both wounding and red light up-regulate the transcript level of ZmAOS, a gene for the jasmonate biosynthesis enzyme allene oxide synthase, and that auxin inhibits the expression of WPK1 but not of ZmAOS. We present a model of jasmonate mediated signaling to explain the results obtained. PMID- 15829514 TI - Nuclear receptors Sf1 and Dax1 function cooperatively to mediate somatic cell differentiation during testis development. AB - Mutations of orphan nuclear receptors SF1 and DAX1 each cause adrenal insufficiency and gonadal dysgenesis in humans, although the pathological features are distinct. Because Dax1 antagonizes Sf1-mediated transcription in vitro, we hypothesized that Dax1 deficiency would compensate for allelic loss of Sf1. In studies of the developing testis, expression of the fetal Leydig cell markers Cyp17 and Cyp11a1 was reduced in heterozygous Sf1-deficient mice at E13.5, consistent with dose-dependent effects of Sf1. In Sf1/Dax1 (Sf1 heterozygous and Dax1-deleted) double mutant gonads, the expression of these genes was unexpectedly reduced further, indicating that loss of Dax1 did not compensate for reduced Sf1 activity. The Sertoli cell product Dhh was reduced in Sf1 heterozygotes at E11.5, and it was undetectable in Sf1/Dax1 double mutants, indicating that Sf1 and Dax1 function cooperatively to induce Dhh expression. Similarly, Amh expression was reduced in both Sf1 and Dax1 single mutants at E11.5, and it was not rescued by the Sf1/Dax1 double mutant. By contrast, Sox9 was expressed in single and in double mutants, suggesting that various Sertoli cell genes are differentially sensitive to Sf1 and Dax1 function. Reduced expression of Dhh and Amh was transient, and was largely restored by E12.5. Similarly, there was recovery of fetal Leydig cell markers by E14.5, indicating that loss of Sf1/Dax1 delays but does not preclude fetal Leydig cell development. Thus, although Sf1 and Dax1 function as transcriptional antagonists for many target genes in vitro, they act independently or cooperatively in vivo during male gonadal development. PMID- 15829515 TI - The ubiquitin ligase Drosophila Mind bomb promotes Notch signaling by regulating the localization and activity of Serrate and Delta. AB - The receptor Notch and its ligands of the Delta/Serrate/LAG2 (DSL) family are the central components in the Notch pathway, a fundamental cell signaling system that regulates pattern formation during animal development. Delta is directly ubiquitinated by Drosophila and Xenopus Neuralized, and by zebrafish Mind bomb, two unrelated RING-type E3 ubiquitin ligases with common abilities to promote Delta endocytosis and signaling activity. Although orthologs of both Neuralized and Mind bomb are found in most metazoan organisms, their relative contributions to Notch signaling in any single organism have not yet been assessed. We show here that a Drosophila ortholog of Mind bomb (D-mib) is a positive component of Notch signaling that is required for multiple Neuralized-independent, Notch dependent developmental processes. Furthermore, we show that D-mib associates physically and functionally with both Serrate and Delta. We find that D-mib uses its ubiquitin ligase activity to promote DSL ligand activity, an activity that is correlated with its ability to induce the endocytosis and degradation of both Delta and Serrate (see also Le Borgne et al., 2005). We further demonstrate that D-mib can functionally replace Neuralized in multiple cell fate decisions that absolutely require endogenous Neuralized, a testament to the highly similar activities of these two unrelated ubiquitin ligases in regulating Notch signaling. We conclude that ubiquitination of Delta and Serrate by Neuralized and D-mib is an obligate feature of DSL ligand activation throughout Drosophila development. PMID- 15829516 TI - The t(8;21) translocation converts AML1 into a constitutive transcriptional repressor. AB - The human translocation (t8;21) is associated with approximately 12% of the cases of acute myelogenous leukemia. Two genes, AML1 and ETO, are fused together at the translocation breakpoint, resulting in the expression of a chimeric protein called AML1-ETO. AML1-ETO is thought to interfere with normal AML1 function, although the mechanism by which it does so is unclear. Here, we have used Drosophila genetics to investigate two models of AML1-ETO function. In the first model, AML1-ETO is a constitutive transcriptional repressor of AML1 target genes, regardless of whether they are normally activated or repressed by AML1. In the second model, AML1-ETO dominantly interferes with AML1 activity by, for example, competing for a common co-factor. To discriminate between these models, the effects of expressing AML1-ETO were characterized and compared with loss-of function phenotypes of lozenge (lz), an AML1 homolog expressed during Drosophila eye development. We also present results of genetic interaction experiments with AML1 co-factors that are not consistent with AML1-ETO behaving as a dominant negative factor. Instead, our data suggest that AML1-ETO acts as a constitutive transcriptional repressor. PMID- 15829517 TI - Production of gurken in the nurse cells is sufficient for axis determination in the Drosophila oocyte. AB - The asymmetric localization of gurken mRNA and protein in the developing Drosophila oocyte defines both the anteroposterior and dorsoventral axes of the future embryo. Understanding the origin of these asymmetries requires knowledge of the source of gurken transcripts. During oogenesis most transcripts in the oocyte are produced by the associated nurse cells, but it has been proposed that gurken is an exceptional oocyte-derived transcript. Using a novel application of a standard mitotic recombination technique, we generated mosaic egg chambers in which the nurse cells, but not the oocyte, could produce gurken. Gurken was properly localized in these mosaics and oocyte axial polarity was established normally, indicating that the nurse cells synthesize gurken and that their contribution is sufficient for Gurken function. Our data demonstrate the existence of a mechanism for transport of gurken from the nurse cells and its subsequent localization within the oocyte. PMID- 15829518 TI - Loss of the extraembryonic ectoderm in Elf5 mutants leads to defects in embryonic patterning. AB - The extraembryonic ectoderm (ExE) is essential for mammalian placental formation and survival of the embryo in utero. We have obtained a mouse model lacking the ExE, by targeted deletion of the transcription factor Elf5. Although Elf5 mutant embryos implant and form an ectoplacental cone, no trophoblast stem (TS) cells can be derived, indicating that the absence of ExE is a result of the lack of TS cell maintenance. Embryos without ExE tissue are able to form the anterior visceral endoderm but fail to undergo gastrulation, demonstrating an essential role for the ExE in embryonic patterning during a defined window of development. PMID- 15829519 TI - Dual roles of zygotic and maternal Scribble1 in neural migration and convergent extension movements in zebrafish embryos. AB - In the developing vertebrate hindbrain, the characteristic trajectory of the facial (nVII) motor nerve is generated by caudal migration of the nVII motor neurons. The nVII motor neurons originate in rhombomere (r) 4, and migrate caudally into r6 to form the facial motor nucleus. In this study, using a transgenic zebrafish line that expresses green fluorescent protein (GFP) in the cranial motor neurons, we isolated two novel mutants, designated landlocked (llk) and off-road (ord), which both show highly specific defects in the caudal migration of the nVII motor neurons. We show that the landlocked locus contains the gene scribble1 (scrb1), and that its zygotic expression is required for migration of the nVII motor neurons mainly in a non cell-autonomous manner. Taking advantage of the viability of the llk mutant embryos, we found that maternal expression of scrb1 is required for convergent extension (CE) movements during gastrulation. Furthermore, we show a genetic interaction between scrb1 and trilobite(tri)/strabismus(stbm) in CE. The dual roles of the scrb1 gene in both neuronal migration and CE provide a novel insight into the underlying mechanisms of cell movement in vertebrate development. PMID- 15829520 TI - Transgenic zebrafish reveal stage-specific roles for Bmp signaling in ventral and posterior mesoderm development. AB - Bone morphogenetic protein (Bmp) signaling is crucial for the formation and patterning of zebrafish ventral and posterior mesoderm. Mutants defective in the Bmp pathway have expanded trunk muscle, abnormal tails and severely impaired development of ventral mesodermal derivatives such as vasculature, blood and pronephros. As Bmps continue to be expressed in the ventral and posterior mesoderm after gastrulation, it is likely that Bmp signaling continues to play an important developmental role during outgrowth of the posterior body. However, because Bmp signaling plays an essential role during the gastrula stages, it has not been possible with mutants or standard disruption techniques to determine the later functions of the Bmp pathway. To study the role of Bmp signaling in the ventral and posterior mesoderm during trunk and tail outgrowth, we generated a transgenic zebrafish line containing a heatshock-inducible dominant-negative Bmp receptor-GFP fusion. Our data show that Bmps are important for tail organizer formation and for patterning the ventral mesoderm during early gastrulation. However, from mid-gastrulation to the early somitogenesis stages, Bmp signaling is important for ventral tail fin development and for preventing secondary tail formation. We conclude that the role of Bmp signaling in the ventral and posterior mesoderm changes as gastrulation proceeds. PMID- 15829521 TI - Gbx2 is required for the morphogenesis of the mouse inner ear: a downstream candidate of hindbrain signaling. AB - Gbx2 is a homeobox-containing transcription factor that is related to unplugged in Drosophila. In mice, Gbx2 and Otx2 negatively regulate each other to establish the mid-hindbrain boundary in the neural tube. Here, we show that Gbx2 is required for the development of the mouse inner ear. Absence of the endolymphatic duct and swelling of the membranous labyrinth are common features in Gbx2-/- inner ears. More severe mutant phenotypes include absence of the anterior and posterior semicircular canals, and a malformed saccule and cochlear duct. However, formation of the lateral semicircular canal and its ampulla is usually unaffected. These inner ear phenotypes are remarkably similar to those reported in kreisler mice, which have inner ear defects attributed to defects in the hindbrain. Based on gene expression analyses, we propose that activation of Gbx2 expression within the inner ear is an important pathway whereby signals from the hindbrain regulate inner ear development. In addition, our results suggest that Gbx2 normally promotes dorsal fates such as the endolymphatic duct and semicircular canals by positively regulating genes such as Wnt2b and Dlx5. However, Gbx2 promotes ventral fates such as the saccule and cochlear duct, possibly by restricting Otx2 expression. PMID- 15829522 TI - Cell cycle diversity involves differential regulation of Cyclin E activity in the Drosophila bristle cell lineage. AB - In the Drosophila bristle lineage, five differentiated cells arise from a precursor cell after a rapid sequence of asymmetric cell divisions (one every 2 hours). We show that, in mitotic cells, this rapid cadence of cell divisions is associated with cell cycles essentially devoid of the G1-phase. This feature is due to the expression of Cyclin E that precedes each cell division, and the differential expression of the S-transition negative regulator, Dacapo. Thus, apart from endocycles (G/S), which occurred in two out of five terminal cells, two other cell cycles coexist in this lineage: (1) an atypical cell cycle (S/G2/M), in which the S-phase is initiated during the preceding telophase; and (2) a canonical cell cycle (G1/S/G2/M) with a brief G1 phase. These two types of cell cycle result from either the absence or very transient expression of Dap, respectively. Finally, we show that the fate determinant factor, Tramtrack, downregulates Cyclin E expression and is probably involved in the exit of the cells from the cell cycle. PMID- 15829523 TI - Neural induction: old problem, new findings, yet more questions. AB - During neural induction, the embryonic neural plate is specified and set aside from other parts of the ectoderm. A popular molecular explanation is the 'default model' of neural induction, which proposes that ectodermal cells give rise to neural plate if they receive no signals at all, while BMP activity directs them to become epidermis. However, neural induction now appears to be more complex than once thought, and can no longer be fully explained by the default model alone. This review summarizes neural induction events in different species and highlights some unanswered questions about this important developmental process. PMID- 15829524 TI - COUP-TFII is essential for radial and anteroposterior patterning of the stomach. AB - COUP-TFII, an orphan member of the steroid receptor superfamily, has been implicated in mesenchymal-epithelial interaction during organogenesis. The generation of a lacZ knock-in allele in the COUP-TFII locus in mice allows us to use X-gal staining to follow the expression of COUP-TFII in the developing stomach. We found COUP-TFII is expressed in the mesenchyme and the epithelium of the developing stomach. Conditional ablation of floxed COUP-TFII by Nkx3-2Cre recombinase in the gastric mesenchyme results in dysmorphogenesis of the developing stomach manifested by major patterning defects in posteriorization and radial patterning. The epithelial outgrowth, the expansion of the circular smooth muscle layer and enteric neurons as well as the posteriorization of the stomach resemble phenotypes exhibited by inhibition of hedgehog signaling pathways. Using organ cultures and cyclopamine treatment, we showed downregulation of COUP-TFII level in the stomach, suggesting COUP-TFII as a target of hedgehog signaling in the stomach. Our results are consistent with a functional link between hedgehog proteins and COUP-TFII, factors that are vital for epithelial-mesenchymal interactions. PMID- 15829525 TI - Nuclear re-organisation of the Hoxb complex during mouse embryonic development. AB - The spatial and temporal co-linear expression of Hox genes during development is an exquisite example of programmed gene expression. The precise mechanisms underpinning this are not known. Analysis of Hoxb chromatin structure and nuclear organisation, during the differentiation of murine ES cells, has lent support to the idea that there is a progressive 'opening' of chromatin structure propagated through Hox clusters from 3'to 5', which contributes to the sequential activation of gene expression. Here, we show that similar events occur in vivo in at least two stages of development. The first changes in chromatin structure and nuclear organisation were detected during gastrulation in the Hoxb1-expressing posterior primitive streak region: Hoxb chromatin was decondensed and the Hoxb1 locus looped out from its chromosome territory, in contrast to non-expressing Hoxb9, which remained within the chromosome territory. At E9.5, when differential Hox expression along the anteroposterior axis is being established, we found concomitant changes in the organisation of Hoxb. Hoxb organisation differed between regions of the neural tube that had never expressed Hoxb [rhombomeres (r) 1 and 2], strongly expressed Hoxb1 but not b9 (r4), had downregulated Hoxb1 (r5), expressed Hoxb9 but not Hoxb1 (spinal cord), and expressed both genes (tail bud). We conclude that Hoxb chromatin decondensation and nuclear re-organisation is regulated in different parts of the developing embryo, and at different developmental stages. The differential nuclear organisation of Hoxb along the anteroposterior axis of the developing neural tube is coherent with co-linear Hox gene expression. In early development nuclear re-organisation is coupled to Hoxb expression, but does not anticipate it. PMID- 15829526 TI - Hym-301, a novel peptide, regulates the number of tentacles formed in hydra. AB - Hym-301 is a peptide that was discovered as part of a project aimed at isolating novel peptides from hydra. We have isolated and characterized the gene Hym-301, which encodes this peptide. In an adult, the gene is expressed in the ectoderm of the tentacle zone and hypostome, but not in the tentacles. It is also expressed in the developing head during bud formation and head regeneration. Treatment of regenerating heads with the peptide resulted in an increase in the number of tentacles formed, while treatment with Hym-301 dsRNA resulted in a reduction of tentacles formed as the head developed during bud formation or head regeneration. The expression patterns plus these manipulations indicate the gene has a role in tentacle formation. Furthermore, treatment of epithelial animals indicates the gene directly affects the epithelial cells that form the tentacles. Raising the head activation gradient, a morphogenetic gradient that controls axial patterning in hydra, throughout the body column results in extending the range of Hym-301 expression down the body column. This indicates the range of expression of the gene appears to be controlled by this gradient. Thus, Hym-301 is involved in axial patterning in hydra, and specifically in the regulation of the number of tentacles formed. PMID- 15829528 TI - Mild cognitive impairment--no benefit from vitamin E, little from donepezil. PMID- 15829527 TI - Vitamin E and donepezil for the treatment of mild cognitive impairment. AB - BACKGROUND: Mild cognitive impairment is a transitional state between the cognitive changes of normal aging and early Alzheimer's disease. METHODS: In a double-blind study, we evaluated subjects with the amnestic subtype of mild cognitive impairment. Subjects were randomly assigned to receive 2000 IU of vitamin E daily, 10 mg of donepezil daily, or placebo for three years. The primary outcome was clinically possible or probable Alzheimer's disease; secondary outcomes were cognition and function. RESULTS: A total of 769 subjects were enrolled, and possible or probable Alzheimer's disease developed in 212. The overall rate of progression from mild cognitive impairment to Alzheimer's disease was 16 percent per year. As compared with the placebo group, there were no significant differences in the probability of progression to Alzheimer's disease in the vitamin E group (hazard ratio, 1.02; 95 percent confidence interval, 0.74 to 1.41; P=0.91) or the donepezil group (hazard ratio, 0.80; 95 percent confidence interval, 0.57 to 1.13; P=0.42) during the three years of treatment. Prespecified analyses of the treatment effects at 6-month intervals showed that as compared with the placebo group, the donepezil group had a reduced likelihood of progression to Alzheimer's disease during the first 12 months of the study (P=0.04), a finding supported by the secondary outcome measures. Among carriers of one or more apolipoprotein E epsilon4 alleles, the benefit of donepezil was evident throughout the three-year follow-up. There were no significant differences in the rate of progression to Alzheimer's disease between the vitamin E and placebo groups at any point, either among all patients or among apolipoprotein E epsilon4 carriers. CONCLUSIONS: Vitamin E had no benefit in patients with mild cognitive impairment. Although donepezil therapy was associated with a lower rate of progression to Alzheimer's disease during the first 12 months of treatment, the rate of progression to Alzheimer's disease after three years was not lower among patients treated with donepezil than among those given placebo. PMID- 15829529 TI - Images in clinical medicine. Airway dilatation after inhalation of a beta agonist. PMID- 15829530 TI - Without conscience. PMID- 15829531 TI - A nutrition paradox--underweight and obesity in developing countries. PMID- 15829532 TI - Marathon maladies. PMID- 15829533 TI - Daily versus as-needed corticosteroids for mild persistent asthma. AB - BACKGROUND: Although guidelines recommend daily therapy for patients with mild persistent asthma, prescription patterns suggest that most such patients use these so-called controller therapies intermittently. In patients with mild persistent asthma, we evaluated the efficacy of intermittent short-course corticosteroid treatment guided by a symptom-based action plan alone or in addition to daily treatment with either inhaled budesonide or oral zafirlukast over a one-year period. METHODS: In a double-blind trial, 225 adults underwent randomization. The primary outcome was morning peak expiratory flow (PEF). Other outcomes included the forced expiratory volume in one second (FEV1) before and after bronchodilator treatment, the frequency of exacerbations, the degree of asthma control, the number of symptom-free days, and the quality of life. RESULTS: The three treatments produced similar increases in morning PEF (7.1 to 8.3 percent; approximately 32 liters per minute; P=0.90) and similar rates of asthma exacerbations (P=0.24), even though the intermittent-treatment group took budesonide, on average, for only 0.5 week of the year. As compared with intermittent therapy or daily zafirlukast therapy, daily budesonide therapy produced greater improvements in pre-bronchodilator FEV1 (P=0.005), bronchial reactivity (P<0.001), the percentage of eosinophils in sputum (P=0.007), exhaled nitric oxide levels (P=0.006), scores for asthma control (P<0.001), and the number of symptom-free days (P=0.03), but not in post-bronchodilator FEV1 (P=0.29) or in the quality of life (P=0.18). Daily zafirlukast therapy did not differ significantly from intermittent treatment in any outcome measured. CONCLUSIONS: It may be possible to treat mild persistent asthma with short, intermittent courses of inhaled or oral corticosteroids taken when symptoms worsen. Further studies are required to determine whether this novel approach to treatment should be recommended. PMID- 15829534 TI - DNA topoisomerase II in therapy-related acute promyelocytic leukemia. AB - BACKGROUND: Chromosomal translocations leading to chimeric oncoproteins are important in leukemogenesis, but how they form is unclear. We studied acute promyelocytic leukemia (APL) with the t(15;17) translocation that developed after treatment of breast or laryngeal cancer with chemotherapeutic agents that poison topoisomerase II. METHODS: We used long-range polymerase chain reaction and sequence analysis to characterize t(15;17) genomic breakpoints in therapy-related APL. To determine whether topoisomerase II was directly involved in mediating breaks of double-stranded DNA at the observed translocation breakpoints, we used a functional in vitro assay to examine topoisomerase II-mediated cleavage in the normal homologues of the PML and RARA breakpoints. RESULTS: Translocation breakpoints in APL that developed after exposure to mitoxantrone, a topoisomerase II poison, were tightly clustered in an 8-bp region within PML intron 6. In functional assays, this "hot spot" and the corresponding RARA breakpoints were common sites of mitoxantrone-induced cleavage by topoisomerase II. Etoposide and doxorubicin also induced cleavage by topoisomerase II at the translocation breakpoints in APL arising after exposure to these agents. Short, homologous sequences in PML and RARA suggested the occurrence of DNA repair by means of the nonhomologous end-joining pathway. CONCLUSIONS: Drug-induced cleavage of DNA by topoisomerase II mediates the formation of chromosomal translocation breakpoints in mitoxantrone-related APL and in APL that occurs after therapy with other topoisomerase II poisons. PMID- 15829535 TI - Hyponatremia among runners in the Boston Marathon. AB - BACKGROUND: Hyponatremia has emerged as an important cause of race-related death and life-threatening illness among marathon runners. We studied a cohort of marathon runners to estimate the incidence of hyponatremia and to identify the principal risk factors. METHODS: Participants in the 2002 Boston Marathon were recruited one or two days before the race. Subjects completed a survey describing demographic information and training history. After the race, runners provided a blood sample and completed a questionnaire detailing their fluid consumption and urine output during the race. Prerace and postrace weights were recorded. Multivariate regression analyses were performed to identify risk factors associated with hyponatremia. RESULTS: Of 766 runners enrolled, 488 runners (64 percent) provided a usable blood sample at the finish line. Thirteen percent had hyponatremia (a serum sodium concentration of 135 mmol per liter or less); 0.6 percent had critical hyponatremia (120 mmol per liter or less). On univariate analyses, hyponatremia was associated with substantial weight gain, consumption of more than 3 liters of fluids during the race, consumption of fluids every mile, a racing time of >4:00 hours, female sex, and low body-mass index. On multivariate analysis, hyponatremia was associated with weight gain (odds ratio, 4.2; 95 percent confidence interval, 2.2 to 8.2), a racing time of >4:00 hours (odds ratio for the comparison with a time of <3:30 hours, 7.4; 95 percent confidence interval, 2.9 to 23.1), and body-mass-index extremes. CONCLUSIONS: Hyponatremia occurs in a substantial fraction of nonelite marathon runners and can be severe. Considerable weight gain while running, a long racing time, and body-mass-index extremes were associated with hyponatremia, whereas female sex, composition of fluids ingested, and use of nonsteroidal antiinflammatory drugs were not. PMID- 15829536 TI - Modification of human hearing loss by plasma-membrane calcium pump PMCA2. AB - Five adult siblings presented with autosomal recessive sensorineural hearing loss: two had high-frequency loss, whereas the other three had severe-to-profound loss affecting all frequencies. Genetic evaluation revealed that a homozygous mutation in CDH23 (which encodes cadherin 23) caused the hearing loss in all five siblings and that a heterozygous, hypofunctional variant (V586M) in plasma membrane calcium pump PMCA2, which is encoded by ATP2B2, was associated with increased loss in the three severely affected siblings. V586M was detected in two unrelated persons with increased sensorineural hearing loss, in the other caused by a mutation in MYO6 (which encodes myosin VI) in one and by noise exposure, suggesting that this variant may modify the severity of sensorineural hearing loss caused by a variety of factors. PMID- 15829537 TI - Effectiveness of antimalarial drugs. PMID- 15829538 TI - Images in clinical medicine. An unusual case of pulmonary embolism. PMID- 15829539 TI - Case records of the Massachusetts General Hospital. Case 11-2005. A 32-year-old pregnant woman with an abnormal fetal karyotype. PMID- 15829540 TI - Does mild persistent asthma require regular treatment? PMID- 15829541 TI - Insights into leukemogenesis from therapy-related leukemia. PMID- 15829542 TI - Resynchronizing ventricular contraction in heart failure. PMID- 15829543 TI - Listening to genetic background noise. PMID- 15829544 TI - Arrest in the liver--a genetically defined malaria vaccine? PMID- 15829545 TI - C-reactive protein levels and outcomes after statin therapy. PMID- 15829546 TI - Molecular prediction of recurrence of breast cancer. PMID- 15829547 TI - Attention deficit-hyperactivity disorder. PMID- 15829548 TI - Prophylaxis against rabies. PMID- 15829549 TI - Case 38-2004: a large tumor of the skull. PMID- 15829550 TI - Distribution of C-reactive protein values in the United States. PMID- 15829551 TI - Clinical trial registration. PMID- 15829552 TI - Altered mental status after a marathon. PMID- 15829553 TI - Feasibility study of a personal dosemeter based upon silicon diodes used in remote controlling. AB - The availability of low cost personal dosemeters is very important for routine worker monitoring. The aim of this work is to evaluate the performance of commercial silicon diodes, commonly used for infrared communication, as a personal radiation monitor. The instrument prototype was characterised both as an area monitor and a personal monitor. Instrument response was measured in the energy range form 37 keV to (60)Co energy. The angular response was also evaluated too. PMID- 15829554 TI - Fraction of the positive 218Po and 214Pb clusters in indoor air. AB - The fraction of the positively charged unattached radon decay products, 218Po and 214Pb in indoor air was determined by model calculations. The results of the calculations were confirmed by measurements in a test chamber (volume: 8 m3). The fraction of both radionuclides depends on the attachment parameter (S(1)) and the neutralisation rate (nu) in room air. The total removal parameter S1 = lambda1 + v + q(f) + X = lambda1 C1f/C0 considers the attachment rate to aerosol particles (X), plate-out rate to room surfaces (q(f)) and the ventilation rate (nu) (lambda1: decay constant of 218Po). The S1-value of room can be determined by measurement of the concentration of the unattached 218Po clusters (C1f) and radon (C0). The neutralisation rate (nu) in environmental air depends mainly on the ion production rate. The influence of the relative humidity in the range 30-95% (temperature: 20 degrees C) is negligible. In addition, equal neutralisation rates for 218Po and 214Pb could be derived. In room air with ion production rates between 5 and 500 nC kg(-1) h(-1) mainly generated by the alpha emitters of radon, thoron and their short-lived decay products, the fractions for positive 218Po clusters vary between 55 and 17% and for 214Pb clusters between 53 and 14%. For a typical average concentration of radon (50 Bq m(-3)) and thoron (10 Bq m( 3)) in homes, 48% of 218Po clusters and 45% of 214Pb clusters are positively charged. PMID- 15829555 TI - The spectrum of Mobius syndrome: an electrophysiological study. AB - We studied the nature and extent of facial muscle innervation and the involvement of the motor and sensory long tracts in Mobius syndrome, in order to shed light on the pathophysiological mechanism of the syndrome. Standardized blink reflexes, direct responses of the facial nerves to the orbicularis oculi muscles and concentric needle electrode electromyography in orbicularis oculi and/or oris muscles were measured in 11 patients with Mobius syndrome, of whom six participated in MRI studies, all showing absent facial nerves. We performed motor and somatosensory-evoked potentials in seven Mobius patients. We demonstrated three distinct patterns of abnormalities suggesting different sites of the primary lesion in different patients. (i) Presence of normal blink reflexes and facial compound motor action potentials, normal habituation tests, a reduced recruitment in the facial muscles and an aberrant 'blink reflex-like' response of the orbicularis oculi muscle upon stimulation of the facial nerve region, which suggests a supranuclear origin of the defect. (ii) Absent blink reflexes, absent direct responses of the facial nerves and absent motor activity on needle electromyography, indicating a defect at the facial nuclear level. However, the nuclear defect might mask an additional supranuclear defect, which cannot, therefore, be excluded in these patients. (iii) A disperse pattern of facial compound action potentials combined with long latencies that were recorded with concentric needle electrodes, indicating involvement of motor axons in the facial nerve, possibly secondary to nuclear involvement. An additional supranuclear defect cannot be excluded in these cases. All evoked potentials studied were normal. The electrophysiological findings of the facial muscles show a spectrum of disturbances varying in degree of severity and diverse in the extent of structures involved, in 11 Mobius patients. At one end of the spectrum are patients with completely immobile faces in whom electrophysiological testing shows no signs of involvement of the facial nuclei, nerves or muscles, suggestive of a dysfunction at the supranuclear level. At the other extreme of the spectrum are patients with complete absence of responses upon facial nerve stimulation and absence of motor unit activity. This is at least indicative of a defect at the facial nuclear level. While a supranuclear defect is compatible with the concept that Mobius syndrome is a developmental disorder of the lower brainstem, intact facial nuclei as part of the syndrome has not been suggested before. The findings corroborate the concept of the Mobius syndrome being a complex regional developmental disorder of the brainstem. PMID- 15829556 TI - Epilepsy surgery does not harm motor performance of children and adolescents. AB - The impact of epilepsy surgery on motor performance, activities of daily life (ADL) and caregiver assistance was assessed in 37 children (age range 0.1-15.4 years) with pharmacologically untreatable epilepsy, 17 of whom were also diagnosed as having spasticity of cerebral origin. All patients underwent epilepsy surgery between 1996 and 2001 at the Wilhelmina University Children's Hospital and were assessed using a standard protocol at fixed intervals: before surgery and 6 months, 1 year and 2 years after surgery. The type of surgery was hemispherectomy (n = 14) and temporal (n = 14), frontal (n = 4), parietal (n = 2) and central (n = 2) resection. One child underwent callosotomy. Engel's classification was used to determine seizure outcome. Impairments were measured in terms of muscle strength, range of motion and muscle tone. Motor performance of infants and children without spasticity was measured using the Movement Assessment Battery for Children (M-ABC). The Gross Motor Function Measure (GMFM 88) was used in children with spasticity, the severity of motor disability in this group being determined by means of the Gross Motor Function Classification System (GMFCS). Daily activities and caregiver's assistance were measured in all children using the Pediatric Evaluation of Disability Inventory (PEDI). Twenty four months after surgery 74% of the children could be classified as Engel class 1, indicating a significant seizure reduction. Impairments revealed some decrease in muscle strength and range of motion in the group with spasticity. Scores improved statistically significantly at group level on M-ABC and GMFM (P < 0.05). Improvement in activities of daily life and caregiver's assistance could not be measured in children without spasticity because of the ceiling effect of the PEDI, but children with spasticity improved significantly with respect to these parameters (PEDI) (P < 0.05). Hence, epilepsy surgery does not harm motor performance in children with or without spasticity. PMID- 15829557 TI - Differentiation- and maturation-dependent content, localization, and secretion of cystatin C in human dendritic cells. AB - Antigen-presenting cells (APC) play a pivotal role in the initiation of the T cell-mediated and antigen-specific immune response. The suggested role of endogenous inhibitor cystatin C (CyC) is to modulate cysteine proteases (cathepsins) present in human APC. To test this hypothesis, dendritic cells (DC) were generated in vitro from isolated monocytes, and changes in content, localization, and secretion of CyC and cathepsins S, L, and H (CatS, -L, and -H, repsectively) were followed in response to interleukin-4, enabling monocyte differentiation, and to tumor necrosis factor alpha (TNF-alpha), enabling DC maturation. A large increase in intracellular CyC accompanied the differentiation of monocytes to immature DC, also shown by strong immunolabeling of Golgi in immature DC. On DC maturation, intracellular CyC levels decreased, and CyC was mostly absent from the Golgi. On prolonged incubation of mature DC with TNF alpha, CyC was found located in the proximity of the plasma membrane, indicating that the transport of CyC from Golgi was not blocked as the result of the arrested exocytosis in mature DC. The secretion of CyC ceased, consistent with the peak of the surface expression of phenotypic markers (CD40, CD54, CD80, CD83, CD86, and major histocompatibility complex class II), characteristic for the mature DC stage, whereas the secretion of cathepsins did not correlate with the maturation stage. The difference in localization of CyC and of CatS, -L, and -H in immature and mature DC shows that the regulatory potential of CyC toward CatS, -L, and -H inside DC is limited. However, these interactions may occur extracellularly in lymph, as suggested by the large excess of CyC over secreted CatS, -L, and -H, and they may facilitate DC migration to lymph nodes. PMID- 15829558 TI - IRF-4 expression in the human myeloid lineage: up-regulation during dendritic cell differentiation and inhibition by 1alpha,25-dihydroxyvitamin D3. AB - Interferon (IFN) regulatory factor (IRF)-4 is a lymphoid- and myeloid-restricted transcription factor of the IRF family. We analyzed its expression during differentiation of human monocytes along the macrophage or the dendritic cell (DC) pathway and in blood myeloid and plasmacytoid DC (M-DC and P-DC, respectively) subsets. Monocyte differentiation into DC, driven by granulocyte macrophage-colony stimulating factor (GM-CSF)/interleukin-4 or GM-CSF/IFN-beta, resulted in a strong up-regulation of IRF-4 mRNA and protein, which was further increased by lipopolysaccharide. It is interesting that 1alpha,25 dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], a potent inhibitor of DC differentiation, completely abolished IRF-4 up-regulation. IRF-4 was also detected in blood P-DC and M-DC. However, up-regulation upon in vitro culture and down-regulation by 1,25(OH)(2)D(3) was observed in M-DC but not in P-DC. These results point to IRF-4 as a potential player in human myeloid DC differentiation and as a novel target for the immunomodulatory activity of 1,25(OH)(2)D(3). PMID- 15829559 TI - Bidirectional regulation of monocyte chemoattractant protein-1 gene at distinct sites of its promoter by nitric oxide in vascular smooth muscle cells. AB - We have previously reported that chronic activation of phosphatidylinositol 3 kinase (PI3-kinase) by the overexpression of membrane-targeted p110CAAX induced proinflammatory gene expression in rat vascular smooth muscle cells (VSMCs) through the induction of CCAAT/enhancer binding protein-beta (C/EBP-beta) and C/EBP-delta. To examine the anti-inflammatory effect of nitric oxide (NO) on proinflammatory gene expression, we have investigated the effects of sodium nitroprusside (SNP) on the monocyte chemoattractant protein-1 (MCP-1) gene expression in VSMCs under chronic activation of PI3-kinase. At low concentrations (0.05 mM) of SNP, but not at high concentrations (0.5-1.0 mM), MCP-1 mRNA and protein expression as well as its transcriptional activity were significantly reduced. We found that SNP induced C/EBP homologous protein (CHOP) expression, which inhibited C/EBP binding activity and reduced the C/EBP activity induced by chronic activation of PI3-kinase in a dose-dependent manner up to 1.0 mM. Consistently, the increase in CHOP expression significantly reduced the MCP-1 promoter activity induced by PI3-kinase. However, the overexpression of CHOP alone upregulated MCP-1 promoter activity in a dose-dependent manner up to high concentrations. Deletion analysis of MCP-1 promoter and electrophoretic mobility shift assay identified the CHOP-response element (CHOP-RE) at the region between 190 and -179 bp of MCP-1 promoter. By using CHOP-RE as a decoy, we significantly suppressed the increase in promoter activity of MCP-1 induced by either CHOP or SNP. Thus CHOP induced by an NO donor has bidirectional effects on MCP-1 gene expression: it decreases gene expression by inhibition of C/EBPs, and it increases the gene expression through CHOP-RE. PMID- 15829560 TI - Inhibition of signal termination-related kinases by membrane-permeant bitter and sweet tastants: potential role in taste signal termination. AB - Sweet and bitter taste sensations are believed to be initiated by the tastant stimulated T1R and T2R G protein-coupled receptor (GPCR) subfamilies, respectively, which occur in taste cells. Although such tastants, with their significantly diverse chemical structures (e.g., sugar and nonsugar sweeteners), may share the same or similar T1Rs, some nonsugar sweeteners and many bitter tastants are amphipathic and produce a significant delay in taste termination (lingering aftertaste). We report that such tastants may permeate rat taste bud cells rapidly in vivo and inhibit known signal termination-related kinases in vitro, such as GPCR kinase (GRK)2, GRK5, and PKA. GRK5 and perhaps GRK2 and GRK6 are present in taste cells. A new hypothesis is proposed in which membrane permeant tastants not only interact with taste GPCRs but also interact intracellularly with the receptors' downstream shutoff components to inhibit signal termination. PMID- 15829561 TI - PYK2 regulates SERCA2 gene expression in neonatal rat ventricular myocytes. AB - The nonreceptor protein tyrosine kinase (PTK) proline-rich tyrosine kinase 2 (PYK2) has been implicated in cell signaling pathways involved in left ventricular hypertrophy and heart failure, but its exact role has not been elucidated. In this study, replication-defective adenoviruses (Adv) encoding green fluorescent protein (GFP)-tagged, wild-type (WT), and mutant forms of PYK2 were used to determine whether PYK2 overexpression activates MAPKs, and downregulates SERCA2 mRNA levels in neonatal rat ventricular myocytes (NRVM). PYK2 overexpression significantly decreased SERCA2 mRNA (as determined by Northern blot analysis and real-time RT-PCR) to 54 +/- 4% of Adv-GFP-infected cells 48 h after Adv infection. Adv-encoding kinase-deficient (KD) and Y(402)F phosphorylation-deficient mutants of PYK2 also significantly reduced SERCA2 mRNA (WT>KD>Y(402)F). Conversely, the PTK inhibitor PP2 (which blocks PYK2 phosphorylation by Src-family PTKs) significantly increased SERCA2 mRNA levels. PYK2 overexpression had no effect on ERK1/2, but increased JNK1/2 and p38(MAPK) phosphorylation from fourfold to eightfold compared with GFP overexpression. Activation of both "stress-activated" protein kinase cascades appeared necessary to reduce SERCA2 mRNA levels. Adv-mediated overexpression of constitutively active (ca)MKK6 or caMKK7, which activated only p38(MAPK) or JNKs, respectively, was not sufficient, whereas combined infection with both Adv reduced SERCA2 mRNA levels to 45 +/- 12% of control. WTPYK2 overexpression also significantly reduced SERCA2 promoter activity, as determined by transient transfection of a 3.8-kb SERCA2 promoter-luciferase construct. Thus a PYK2-dependent signaling cascade may have a role in abnormal cardiac Ca(2+) handling in left ventricular hypertrophy and heart failure via downregulation of SERCA2 gene transcription. PMID- 15829562 TI - Proton-sensing G protein-coupled receptor mobilizes calcium in human synovial cells. AB - Lowered extracellular pH in a variety of tissues is associated with increased tissue destruction and initiation of inflammatory processes. Although the acid sensing receptors described previously are ion channels, we describe a G protein coupled proton-sensitive receptor that stimulates Ca(2+) release from intracellular stores in a tumor-derived synoviocyte cell line (SW982) and in primary cultures of human synovial cells from patients with inflammatory arthropathies. We established a link between proton-dependent receptor activation and intracellular Ca(2+) mobilization by demonstrating 1) dependence on the integrity of the intracellular Ca(2+) store, 2) independence from extracellular Ca(2+), and 3) proton-induced production of inositol phosphate and 4) by abolishing the effect with GTPase inhibitors. We propose that this G protein coupled acid-sensing receptor linked to intracellular Ca(2+) mobilization in synoviocytes can contribute to downstream inflammatory and cellular proliferative processes in synovial fibroblasts. The acid-sensing receptor has distinct characteristics as a metabotropic G protein-coupled receptor on human synoviocytes in this emerging new class of receptors. PMID- 15829563 TI - Caspase-resistant Golgin-160 disrupts apoptosis induced by secretory pathway stress and ligation of death receptors. AB - Golgin-160 is a coiled-coil protein on the cytoplasmic face of the Golgi complex that is cleaved by caspases during apoptosis. We assessed the sensitivity of cell lines stably expressing wild-type or caspase-resistant golgin-160 to several proapoptotic stimuli. Cells expressing a caspase-resistant mutant of golgin-160 were strikingly resistant to apoptosis induced by ligation of death receptors and by drugs that induce endoplasmic reticulum (ER) stress, including brefeldin-A, dithiothreitol, and thapsigargin. However, both cell lines responded similarly to other proapoptotic stimuli, including staurosporine, anisomycin, and etoposide. The caspase-resistant golgin-160 dominantly prevented cleavage of endogenous golgin-160 after ligation of death receptors or induction of ER stress, which could be explained by a failure of initiator caspase activation. The block in apoptosis in cells expressing caspase-resistant golgin-160 could not be bypassed by expression of potential caspase cleavage fragments of golgin-160, or by drug induced disassembly of the Golgi complex. Our results suggest that some apoptotic signals (including those initiated by death receptors and ER stress) are sensed and integrated at Golgi membranes and that golgin-160 plays an important role in transduction of these signals. PMID- 15829564 TI - Microtubules are dispensable for the initial pathogenic development but required for long-distance hyphal growth in the corn smut fungus Ustilago maydis. AB - Fungal pathogenicity often involves a yeast-to-hypha transition, but the structural basis for this dimorphism is largely unknown. Here we analyze the role of the cytoskeleton in early steps of pathogenic development in the corn pathogen Ustilago maydis. On the plant yeast-like cells recognize each other, undergo a cell cycle arrest, and form long conjugation hyphae, which fuse and give rise to infectious filaments. F-actin is essential for polarized growth at all these stages and for cell-cell fusion. Furthermore, F-actin participates in pheromone secretion, but not perception. Although U. maydis contains prominent tubulin arrays, microtubules are neither required for cell-cell recognition, nor for cell cell fusion, and have only minor roles in morphogenesis of yeast-like cells. Without microtubules hyphae are formed, albeit at 60% reduced elongation rates, but they reach only approximately 50 mum in length and the nucleus fails to migrate into the hypha. A similar phenotype is found in dynein mutants that have a nuclear migration defect and stop hyphal elongation at approximately 50 mum. These results demonstrate that microtubules are dispensable for polarized growth during morphological transition, but become essential in long-distance hyphal growth, which is probably due to their role in nuclear migration. PMID- 15829565 TI - Dynein light chain 1 regulates dynamin-mediated F-actin assembly during sperm individualization in Drosophila. AB - Toward the end of spermiogenesis, spermatid nuclei are compacted and the clonally related spermatids individualize to become mature and active sperm. Studies in Drosophila showed that caudal end-directed movement of a microfilament-rich structure, called investment cone, expels the cytoplasmic contents of individual spermatids. F-actin dynamics plays an important role in this process. Here we report that the dynein light chain 1 (DLC1) of Drosophila is involved in two separate cellular processes during sperm individualization. It is enriched around spermatid nuclei during postelongation stages and plays an important role in the dynein-dynactin-dependent rostral retention of the nuclei during this period. In addition, DDLC1 colocalizes with dynamin along investment cones and regulates F actin assembly at this organelle by retaining dynamin along the cones. Interestingly, we found that this process does not require the other subunits of cytoplasmic dynein-dynactin complex. Altogether, these observations suggest that DLC1 could independently regulate multiple cellular functions and established a novel role of this protein in F-actin assembly in Drosophila. PMID- 15829566 TI - The conserved Mec1/Rad53 nuclear checkpoint pathway regulates mitochondrial DNA copy number in Saccharomyces cerevisiae. AB - How mitochondrial DNA (mtDNA) copy number is determined and modulated according to cellular demands is largely unknown. Our previous investigations of the related DNA helicases Pif1p and Rrm3p uncovered a role for these factors and the conserved Mec1/Rad53 nuclear checkpoint pathway in mtDNA mutagenesis and stability in Saccharomyces cerevisiae. Here, we demonstrate another novel function of this pathway in the regulation of mtDNA copy number. Deletion of RRM3 or SML1, or overexpression of RNR1, which recapitulates Mec1/Rad53 pathway activation, resulted in an approximately twofold increase in mtDNA content relative to the corresponding wild-type yeast strains. In addition, deletion of RRM3 or SML1 fully rescued the approximately 50% depletion of mtDNA observed in a pif1 null strain. Furthermore, deletion of SML1 was shown to be epistatic to both a rad53 and an rrm3 null mutation, placing these three genes in the same genetic pathway of mtDNA copy number regulation. Finally, increased mtDNA copy number via the Mec1/Rad53 pathway could occur independently of Abf2p, an mtDNA-binding protein that, like its metazoan homologues, is implicated in mtDNA copy number control. Together, these results indicate that signaling through the Mec1/Rad53 pathway increases mtDNA copy number by altering deoxyribonucleoside triphosphate pools through the activity of ribonucleotide reductase. This comprises the first linkage of a conserved signaling pathway to the regulation of mitochondrial genome copy number and suggests that homologous pathways in humans may likewise regulate mtDNA content under physiological conditions. PMID- 15829567 TI - Nuclear import of the stem-loop binding protein and localization during the cell cycle. AB - A key factor involved in the processing of histone pre-mRNAs in the nucleus and translation of mature histone mRNAs in the cytoplasm is the stem-loop binding protein (SLBP). In this work, we have investigated SLBP nuclear transport and subcellular localization during the cell cycle. SLBP is predominantly nuclear under steady-state conditions and localizes to the cytoplasm during S phase when histone mRNAs accumulate. Consistently, SLBP mutants that are defective in histone mRNA binding remain nuclear. As assayed in heterokaryons, export of SLBP from the nucleus is dependent on histone mRNA binding, demonstrating that SLBP on its own does not possess any nuclear export signals. We find that SLBP interacts with the import receptors Impalpha/Impbeta and Transportin-SR2. Moreover, complexes formed between SLBP and the two import receptors are disrupted by RanGTP. We have further shown that SLBP is imported by both receptors in vitro. Three sequences in SLBP required for Impalpha/Impbeta binding were identified. Simultaneous mutation of all three sequences was necessary to abolish SLBP nuclear localization in vivo. In contrast, we were unable to identify an in vivo role for Transportin-SR2 in SLBP nuclear localization. Thus, only the Impalpha/Impbeta pathway contributes to SLBP nuclear import in HeLa cells. PMID- 15829568 TI - The membrane-anchoring domain of epidermal growth factor receptor ligands dictates their ability to operate in juxtacrine mode. AB - All ligands of the epidermal growth factor (EGF) receptor (EGFR) are synthesized as membrane-anchored precursors. Previous work has suggested that some ligands, such as EGF, must be proteolytically released to be active, whereas others, such as heparin-binding EGF-like growth factor (HB-EGF) can function while still anchored to the membrane (i.e., juxtacrine signaling). To explore the structural basis for these differences in ligand activity, we engineered a series of membrane-anchored ligands in which the core, receptor-binding domain of EGF was combined with different domains of both EGF and HB-EGF. We found that ligands having the N-terminal extension of EGF could not bind to the EGFR, even when released from the membrane. Ligands lacking an N-terminal extension, but possessing the membrane-anchoring domain of EGF, still required proteolytic release for activity, whereas ligands with the membrane-anchoring domain of HB EGF could elicit full biological activity while still membrane anchored. Ligands containing the HB-EGF membrane anchor, but lacking an N-terminal extension, activated EGFR during their transit through the Golgi apparatus. However, cell mixing experiments and fluorescence resonance energy transfer studies showed that juxtacrine signaling typically occurred in trans at the cell surface, at points of cell-cell contact. Our data suggest that the membrane-anchoring domain of ligands selectively controls their ability to participate in juxtacrine signaling and thus, only a subclass of EGFR ligands can act in a juxtacrine mode. PMID- 15829569 TI - Coronin-1 function is required for phagosome formation. AB - Coronin-1 is an actin-associated protein whose function in actin dynamics has remained obscure. All coronin proteins have a variable N-terminal domain, followed by WD repeats and a C-terminal coiled-coil dimerization domain. Transfection of coronin-1-GFP into RAW 264.7 cells revealed that coronin rapidly and transiently associates with the phagosome. To determine if coronin is involved in mammalian phagocytosis we used a dominant-negative approach by expressing only the central WD domains. However, this caused cell rounding and dissociation from the substratum, hampering analysis of their phenotype. We therefore developed TAT-fusion constructs of coronin-1 WD domains to acutely introduce the recombinant protein fragment into live cells. We show that although TAT-WD has no effect on binding of opsonized RBCs to RAW 264.7 cells, receptor clustering or several downstream signaling events, lamellipodial extensions, and actin accumulation at the base of the bound particle were diminished. Furthermore, Arp3 accumulation at the phagosome was impaired after TAT-WD treatment. Interestingly, whereas coronin-1 also accumulates at the sites of actin remodeling associated with Salmonella invasion, TAT-WD had no effect on this process. Together, our data demonstrates that coronin-1 is required for an early step in phagosome formation, consistent with a role in actin polymerization. PMID- 15829570 TI - Impairment of the TFIIH-associated CDK-activating kinase selectively affects cell cycle-regulated gene expression in fission yeast. AB - The fission yeast Mcs6-Mcs2-Pmh1 complex, homologous to metazoan Cdk7-cyclin H Mat1, has dual functions in cell division and transcription: as a partially redundant cyclin-dependent kinase (CDK)-activating kinase (CAK) that phosphorylates the major cell cycle CDK, Cdc2, on Thr-167; and as the RNA polymerase (Pol) II carboxyl-terminal domain (CTD) kinase associated with transcription factor (TF) IIH. We analyzed conditional mutants of mcs6 and pmh1, which activate Cdc2 normally but cannot complete cell division at restrictive temperature and arrest with decreased CTD phosphorylation. Transcriptional profiling by microarray hybridization revealed only modest effects on global gene expression: a one-third reduction in a severe mcs6 mutant after prolonged incubation at 36 degrees C. In contrast, a small subset of transcripts ( approximately 5%) decreased by more than twofold after Mcs6 complex function was compromised. The signature of repressed genes overlapped significantly with those of cell separation mutants sep10 and sep15. Sep10, a component of the Pol II Mediator complex, becomes essential in mcs6 or pmh1 mutant backgrounds. Moreover, transcripts dependent on the forkhead transcription factor Sep1, which are expressed coordinately during mitosis, were repressed in Mcs6 complex mutants, and Mcs6 also interacts genetically with Sep1. Thus, the Mcs6 complex, a direct activator of Cdc2, also influences the cell cycle transcriptional program, possibly through its TFIIH-associated kinase function. PMID- 15829571 TI - Activity limitations and participation restrictions in women with hand osteoarthritis: patients' descriptions and associations between dimensions of functioning. AB - OBJECTIVE: To describe the functional consequences of hand osteoarthritis, and analyse associations between personal factors, hand impairment, activity limitations, and participation restrictions within the framework of the International Classification of Functioning (ICF). METHODS: 87 women with hand osteoarthritis completed a clinical examination including recording of sociodemographic data, measures of hand impairment, and completion of self reported health status measures. The function subscale of the AUSCAN Osteoarthritis Hand Index was used as a measure of hand related activity limitations, while the Canadian Occupational Performance Measure (COPM) was used to describe and measure activity limitations and participation restrictions as perceived by the individual. The study variables were categorised using the dimensions in the ICF framework and analysed using bivariate and multivariate statistical approaches. RESULTS: The patients described problems in many domains of activity and participation. The most frequently described hand related problems were activities requiring considerable grip strength combined with twisting of the hands. On the impairment level, the patients had reduced grip force and joint mobility in the hands, and resisted motion was painful. Regression analyses showed that hand related activity limitations were associated with measures of hand impairment, while activity and participation (as measured by the COPM) were more strongly associated with personal factors than with hand impairment. CONCLUSIONS: Hand osteoarthritis has important functional consequences in terms of pain, reduced hand mobility and grip force, activity limitations, and participation restrictions. Rehabilitation programmes should therefore be multidisciplinary and multidimensional, focusing on hand function, occupational performance, and coping strategies. PMID- 15829572 TI - Risks of solid cancers in patients with rheumatoid arthritis and after treatment with tumour necrosis factor antagonists. AB - BACKGROUND: Existing studies of solid cancers in rheumatoid arthritis (RA) reflect cancer morbidity up until the early 1990s in prevalent cohorts admitted to hospital during the 1980s. OBJECTIVE: To depict the cancer pattern of contemporary patients with RA, from updated risk data from prevalent and incident RA populations. To understand the risk of solid cancer after tumour necrosis factor (TNF) treatment by obtaining cancer data from cohorts treated in routine care rather than trials. METHODS: A population based study of three RA cohorts (one prevalent, admitted to hospital 1990-2003 (n = 53,067), one incident, diagnosed 1995-2003 (n = 3703), and one treated with TNF antagonists 1999-2003 (n = 4160)), which were linked with Swedish nationwide cancer and census registers and followed up for cancer occurrence through 2003. RESULTS: With 3379 observed cancers, the prevalent RA cohort was at marginally increased overall risk of solid cancer, with 20-50% increased risks for smoke related cancers and +70% increased risk for non-melanoma skin cancer, but decreased risk for breast (-20%) and colorectal cancer (-25%). With 138 cancers, the incident RA cohort displayed a similar cancer pattern apart from non-decreased risks for colorectal cancer. TNF antagonist treated patients displayed solid cancer (n = 67) risks largely similar to those of other patients with RA. CONCLUSION: The cancer pattern in patients treated with TNF antagonists mirrors those of other contemporary as well as historic RA cohorts. The consistent increase in smoking associated cancers in patients with RA emphasises the potential for smoking cessation as a cancer preventive measure in RA. PMID- 15829573 TI - Outcome of intensive immunosuppression and autologous stem cell transplantation in patients with severe rheumatoid arthritis is associated with the composition of synovial T cell infiltration. AB - OBJECTIVE: To determine clinical and immunological correlates of high dose chemotherapy (HDC) + autologous stem cell transplantation (ASCT) in patients with severe rheumatoid arthritis (RA), refractory to conventional treatment. METHODS: Serial samples of peripheral blood and synovial tissue were obtained from seven patients with RA treated with HDC and autologous peripheral blood grafts enriched for CD34+ cells. Disease activity was assessed with the Disease Activity Score (DAS), serum concentrations of C reactive protein (CRP), and human immunoglobulin (HIg) scans, and the extent of immunoablation was determined by immunophenotyping of peripheral blood mononuclear cells, and immunohistochemistry and double immunofluorescence of synovium. RESULTS: Clinical responders (n = 5) had a larger number of cells at baseline expressing CD3, CD4, CD27, CD45RA, CD45RB, and CD45RO in synovium (p < 0.05), higher activity on HIg scans (p = 0.08), and a trend towards higher concentrations of CRP in serum than non-responders (n = 2). Subsequent remissions and relapses in responders paralleled reduction and re expression, respectively, of T cell markers. A relatively increased expression of CD45RB and CD45RO on synovial CD3+ T cells was seen after HDC + ASCT. No correlations were found between DAS and changes in B cells or macrophage infiltration or synoviocytes. CONCLUSIONS: HDC + ASCT results in profound but incomplete immunoablation of both the memory and naive T cell compartment, which is associated with longlasting clinical responses in most patients. The findings provide strong circumstantial evidence for a role of T cells in established RA, and demonstrate a role for the synovium in post-transplantation T cell reconstitution. PMID- 15829574 TI - Effectiveness of systematic monitoring of rheumatoid arthritis disease activity in daily practice: a multicentre, cluster randomised controlled trial. AB - OBJECTIVE: To test the efficacy of standardised monitoring using the disease activity index DAS28 versus usual care on disease modifying antirheumatic drug (DMARD) prescription and disease activity in rheumatoid arthritis. METHODS: A 24 week cluster randomised trial. Rheumatology outpatient centres were randomised to systematic monitoring of disease activity using the DAS28 (12 centres, 205 patients) or usual care (12 centres, 179 patients). The aim for the DAS group was to reach a DAS28 score of < or =3.2 by changes in DMARD treatment, at the discretion of the rheumatologist and the patient. RESULTS: At baseline, disease activity was the same in both groups, with an overall mean (SD) DAS28 of 4.5 (1.2); 13% of the patients had a DAS28 of < or =3.2. At 24 weeks, 31% of patients in the DAS group had a DAS28 < or =3.2, while in the usual care centres this was 16% (p = 0.028). DMARDs were changed on average in 18% of visits in the DAS centres; in the 12 usual care centres they were changed on 8% of the visits (p = 0.013). The doses of methotrexate, sulfasalazine, and corticosteroids appeared to be higher in the DAS centres than in the usual care centres, but the differences were not significant. CONCLUSIONS: In daily practice, systematic monitoring of disease activity in rheumatoid arthritis may lead to more changes in DMARD treatment, resulting in a larger number of patients with low disease activity. PMID- 15829575 TI - Improvement of coping abilities in patients with systemic lupus erythematosus: a prospective study. AB - OBJECTIVE: To evaluate a novel specific psychological intervention aimed at improving coping in patients with systemic lupus erythematosus (SLE). METHODS: 34 community living SLE patients were recruited for the study. Intervention was undertaken in groups of up to eight patients and in two blocks over six months each. Eight patients were enrolled as a waiting list group. The 18 group sessions focused on information about the disease and specific problems of SLE patients, combining psychoeducative and psychotherapeutic elements. Psychological and medical evaluations were conducted at baseline and after three, six, and 12 months, using validated instruments. RESULTS: The 34 SLE patients (91% female, mean age 42 years) improved significantly over a six month period on most of the psychological measuring instruments applied, such as depression, anxiety, and overall mental burden. The waiting list group showed no significant changes. CONCLUSIONS: Conceptualised psychoeducational support may produce a significant and sustained improvement in coping skills of SLE patients and hence in their quality of life. PMID- 15829576 TI - Hard work never hurt anyone--or did it? A review of occupational associations with soft tissue musculoskeletal disorders of the neck and upper limb. AB - Pain in the neck and upper limb is common and contributes greatly to absence due to sickness. Evidence is accumulating that factors such as prolonged abnormal posture and repetition contribute to these conditions. Psychosocial factors may also play a part and the relative importance of these is not yet known. Primary and secondary prevention trials are needed. PMID- 15829577 TI - Infliximab in combination with methotrexate in active ankylosing spondylitis: a clinical and imaging study. AB - OBJECTIVE: To examine the efficacy and safety of infliximab combined with methotrexate compared with methotrexate alone in the treatment of ankylosing spondylitis (AS) using MRI and DXA to monitor its impact on bone. METHODS: In this single centre study 42 subjects with active AS were treated with methotrexate and were randomly assigned, in a ratio of 2:1, to receive five infusions of either 5 mg/kg infliximab or placebo over 30 weeks. The primary outcome was improvement in disease activity as shown by the BASDAI at week 30. MRI was used to assess the effect of treatments on sacroiliac and spinal enthesitis/osteitis and DXA to monitor bone mineral density. RESULTS: Both therapeutic agents were well tolerated with no dropouts due to adverse events. A significantly greater improvement in mean BASDAI score was seen in the infliximab arm at week 10 (p = 0.017) than in the placebo arm, but this was not maintained by week 30 (p = 0.195), 8 weeks after the last infusion, at which stage disease flares were reported by some subjects. MRI showed that the mean number of lesions resolving for each subject from week 0 to week 30 was significantly greater in the combination group than in the methotrexate monotherapy group (p = 0.016). CONCLUSIONS: Infliximab in combination with methotrexate was a safe and efficacious treatment in AS over 6 months and was associated with significant regression in enthesitis/osteitis as determined by MRI. However, disease flares were reported 8 weeks after the last infusion, indicating that addition of methotrexate failed to extend the infliximab dosing interval. PMID- 15829578 TI - Altered cytokine expression of peripheral blood lymphocytes in polymyositis and dermatomyositis. AB - OBJECTIVE: To investigate the intracellular and soluble cytokine levels and T cell subsets in peripheral blood of patients with active and inactive polymyositis and dermatomyositis. METHODS: The frequencies of T and B lymphocytes, T helper (Th), and T cytotoxic (Tc) cells and of interferon gamma (IFNgamma), interleukin (IL)4, and IL10 expression of CD4+ or CD8+ cells were determined by flow cytometry. The concentrations of soluble cytokines were measured with commercial enzyme linked immunosorbent assays. RESULTS: In active dermatomyositis there was a decreased percentage of T (CD3+) lymphocytes and Tc (CD8+) lymphocytes, decreased IFNgamma expression of CD4+ and CD8+ cells, but an increase in B and IL4 producing CD4+ lymphocyte frequencies. These prominent changes disappeared in the inactive stage of the disease. In polymyositis no significant change in these lymphocyte subsets or in intracellular cytokine expression could be detected in either the active or the inactive form. The frequency of IL4+/IFNgamma+ Th cells was calculated and a significantly increased Th2/Th1 frequency was found in active dermatomyositis, and a decreased frequency in inactive dermatomyositis, compared with the control population. CONCLUSIONS: There appears to be a difference between polymyositis and dermatomyositis in the level of peripheral blood lymphocytes and their intracellular cytokine content. These findings provide further evidence for a difference in the pathogenesis of polymyositis and dermatomyositis. PMID- 15829579 TI - Presence of NGF and its receptors in ovaries from human fetuses and adults. AB - The ability to mature human primordial follicles in vitro would assist fertility restoration. However, the signals initiating growth of primordial follicles are unknown. Growth factors such as nerve growth factor (NGF) may play a role in this process. To investigate the expression of NGF and its receptors, p75 and TrkA, in early developing follicles (mostly primordial, primary and secondary follicles), ten ovarian samples from adolescents/adults aged 13-39 and 33 ovaries from human fetuses aged 19-33 gestational weeks (GW) were obtained and immediately fixed or frozen. The fixed samples were prepared for a study of immunocytochemical staining of NGF and its two receptors. Total RNA was extracted from the frozen ovarian samples, and the expression of NGF, TrkA and p75 was investigated by RT PCR. Products were resolved by 1% agarose gel electrophoresis and image analysis. Immunocytochemical staining revealed the expression of NGF in granulosa cells (GC) and oocytes; TrkA was mainly in oocytes and in GC in minority of the samples; and p75 was in some of the stroma cells from fetuses aged less than 22 GW. Transcripts of NGF and TrkA were identified by RT-PCR in all samples, while those for p75 were detected only in ovarian samples from fetuses aged less than 22 GW. To elucidate if NGF is indeed involved in growth initiation of human primordial follicles, it should be added to their culture medium. The immunocytochemical detection of p75 in some of the stroma cells and transcripts in ovarian samples of fetuses less than 22 GW may suggest its role in follicular assembly. PMID- 15829580 TI - Patterns of expression of sperm flagellar genes: early expression of genes encoding axonemal proteins during the spermatogenic cycle and shared features of promoters of genes encoding central apparatus proteins. AB - Sperm are motile cells. Thus, a significant component of the spermatogenic cycle is devoted to the formation of flagellum, a process that must be coordinated to insure proper construction. To document the temporal pattern of flagellar gene expression, we employed real-time PCR to assess changes in accumulation of a cohort of genes encoding axoneme, outer dense fibre (ODF) and fibrous sheath (FS) proteins during the first wave of spermatogenesis in the mouse. Axoneme genes were expressed first at the pachytene spermatocyte stage, followed by expression of transcripts encoding ODF and FS components. However, there were differences among these families with respect to the time of initial expression and the rate of mRNA accumulation. To gain understanding of factors that determine these patterns of expression, we cloned the promoters of three axoneme central apparatus genes (Pf6, Spag6 and Pf20). These promoters shared common features including the absence of a TATA box, and putative binding sites for several factors implicated in spermatogenesis (CREB/CREM, SOX17 and SPZ1) as well as ciliogenesis (FOXJ1). Collectively, our findings demonstrate a sequential pattern of expression of flagellar component genes, differential times of expression or rates of transcript accumulation within each class and shared promoter features within a class. PMID- 15829581 TI - Global archaeological evidence for proboscidean overkill. AB - One million years ago, proboscideans occupied most of Africa, Europe, Asia, and the Americas. Today, wild elephants are only found in portions of sub-Saharan Africa and South Asia. Although the causes of global Pleistocene extinctions in the order Proboscidea remain unresolved, the most common explanations involve climatic change and/or human hunting. In this report, we test the overkill and climate-change hypotheses by using global archaeological spatiotemporal patterning in proboscidean kill/scavenge sites. Spanning approximately 1.8 million years, the archaeological record of human subsistence exploitation of proboscideans is preferentially located on the edges of the human geographic range. This finding is commensurate with global overkill, suggesting that prehistoric human range expansion resulted in localized extinction events. In the present and the past, proboscideans have survived in refugia that are largely inaccessible to human populations. PMID- 15829582 TI - Protein fibrils in nature can enhance amyloid protein A amyloidosis in mice: Cross-seeding as a disease mechanism. AB - Secondary, or amyloid protein A (AA), amyloidosis is a complication of chronic inflammatory diseases, both infectious and noninfectious. AA constitutes the insoluble fibrils, which are deposited in different organs, and is a major N terminal part of the acute phase protein serum AA. It is not known why only some patients with chronic inflammation develop AA amyloidosis. Nucleation is a widely accepted mechanism in amyloidogenesis. Preformed amyloid-like fibrils act as nuclei in amyloid fibril formation in vitro, and AA amyloid fibrils and synthetic amyloid-like fibrils also may serve as seed for fibril formation in vivo. In addition to amyloid fibrils, there is a variety of similar nonmammalian protein fibrils with beta-pleated structure in nature. We studied three such naturally occurring protein fibrils: silk from Bombyx mori, Sup35 from Saccharomyces cerevisiae, and curli from Escherichia coli. Our results show that these protein fibrils exert amyloid-accelerating properties in the murine experimental AA amyloidosis, suggesting that such environment factors may be important risk factors in amyloidogenesis. PMID- 15829584 TI - Germ cell mutagenesis in medaka fish after exposures to high-energy cosmic ray nuclei: A human model. AB - Astronauts beyond the Earth's orbit are exposed to high-energy cosmic-ray nuclei with high values of linear energy transfer (LET), resulting in much more biological damage than from x-rays or gamma-rays and may result in mutations and cancer induction. The relative biological effectiveness of these nuclei depends on the LET, rising to as high as approximately 50 at LET values of approximately 100-200 keV/microm. An endpoint of concern is germ cell mutations passed on to offspring, arising from exposure to these nuclei. A vertebrate model for germ cell mutation is Medaka fish (Oryzias latipes). We exposed wild type males to doses of 1 GeV per nucleon Fe nuclei or to 290 MeV per nucleon C nuclei. They were mated to females with recessive mutations at five-color loci. The transparent embryos from >100 days of mating (representing exposed sperm, spermatids, or spermatogonia) were observed so as to detect dominant lethal mutations and total color mutations, even though the embryos might not hatch. The relative number of mutant embryos as a function of dose were compared with those induced by gamma-rays. The relative biological effectiveness values for dominant lethal mutations and total color mutations for exposed sperm and spermatids were 1.3-2.1 for exposure to C nuclei and 1.5-3.0 for exposure to Fe nuclei. (The spermatogonial data were uncertain.) These low values, and the negligible number of viable mutations, compared with those for mutations in somatic cells and for neoplastic transformation, indicate that germ cell mutations arising from exposures to cosmic ray nuclei are not a significant hazard to astronauts. PMID- 15829583 TI - Zinc inhibition of gamma-aminobutyric acid transporter 4 (GAT4) reveals a link between excitatory and inhibitory neurotransmission. AB - gamma-Aminobutyric acid (GABA) transporters (GATs) play an important role in inhibitory neurotransmission by clearing synaptically released GABA and by maintaining low resting levels of GABA in synaptic and extrasynaptic regions. In certain brain regions, vesicular zinc is colocalized and coreleased with glutamate and modulates the behavior of a number of channels, receptors, and transporters. We examined the effect of zinc on expressed GATs (GAT1, GAT2, GAT3, and GAT4) in Xenopus laevis oocytes by using tracer flux and electrophysiological methods. We show that zinc is a potent inhibitor of GAT4 (K(i) of 3 muM). Immunolocalization of GAT4 in the hippocampus revealed dense localization in the CA1 and CA3 regions of the hippocampus, regions which are known to be heavily populated by zinc-containing glutamatergic neurons. The results suggest a physiological role of synaptically released zinc in the hippocampus, because zinc released from hyperactive glutamatergic neurons may simultaneously bring about elevated GABAergic inhibition. Therefore, this mode of zinc function signifies a link between excitatory and inhibitory neurotransmission and may play a neuroprotective role against glutamate-induced excitotoxicity. PMID- 15829585 TI - Meat and milk compositions of bovine clones. AB - The technology is now available for commercial cloning of farm animals for food production, but is the food safe for consumers? Here, we provide data on >100 parameters that compare the composition of meat and milk from beef and dairy cattle derived from cloning to those of genetic- and breed-matched control animals from conventional reproduction. The cloned animals and the comparators were managed under the same conditions and received the same diet. The composition of the meat and milk from the clones were largely not statistically different from those of matched comparators, and all parameters examined were within the normal industry standards or previously reported values. The data generated from our match-controlled experiments provide science-based information desired by regulatory agencies to address public concerns about the safety of meat and milk from somatic animal clones. PMID- 15829586 TI - Rare creation of recombinant mtDNA haplotypes in mammalian tissues. AB - The problem of whether recombinant mtDNAs are created in mammalian cells has been controversial for many years. We show convincing evidence for the very rare creation of recombinant mtDNA haplotypes by isolating human somatic hybrid cells and by generating mice carrying two different mtDNA haplotypes. To avoid misinterpretation of PCR-jumping products as recombinants, we used purified mtDNAs for cloning and sequencing. The results showed that only three of 318 clones of mtDNA purified from mouse tissues corresponded to recombinant mtDNA haplotypes, whereas no recombinants were found in human somatic hybrid cells. Such an extremely low frequency of mtDNA recombination does not require any revision of important concepts on human evolution that are based on its absence. Considering the high concentration of reactive oxygen species around the mtDNA and its frequent strand breakage, recombinant clones would correspond to gene conversion products created by repair of nucleotide mismatches. PMID- 15829587 TI - Self-limiting nature of seasonal cholera epidemics: Role of host-mediated amplification of phage. AB - Phage predation of Vibrio cholerae has recently been reported to be a factor that influences seasonal epidemics of cholera in Bangladesh. To understand more about this phenomenon, we studied the dynamics of the V. cholerae-phage interaction during a recent epidemic in Dhaka. Because the outbreak strain causing this epidemic was resistant to multiple antibiotics, including streptomycin, we used a selective medium containing streptomycin to monitor accurately the abundance of this strain in the environment. The changing prevalence in the environment of the epidemic V. cholerae O1 strain and a particular lytic cholera phage (JSF4) to which it was sensitive was measured every 48-72 h for 17 weeks. We also monitored the incidence of phage excretion in stools of 387 cholera patients during the epidemic. The peak of the epidemic was preceded by high V. cholerae prevalence in the environment and was followed by high JSF4 phage levels as the epidemic ended. The buildup to the phage peak in the environment coincided with increasing excretion of the same phage in the stools of cholera patients. These results suggest that patients toward the end of the epidemic ingested both JSF4 phage and the outbreak V. cholerae strain. Host-mediated phage amplification during the cholera epidemic likely contributed to increased environmental phage abundance, decreased load of environmental V. cholerae and, hence, the collapse of the epidemic. Thus, in vivo phage amplification in patients and subsequent phage predation in the environment may explain the self-limiting nature of seasonal cholera epidemics in Bangladesh. PMID- 15829588 TI - Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides. AB - We report four-color DNA sequencing by synthesis (SBS) on a chip, using four photocleavable fluorescent nucleotide analogues (dGTP-PC-Bodipy-FL-510, dUTP-PC R6G, dATP-PC-ROX, and dCTP-PC-Bodipy-650) (PC, photocleavable; Bodipy, 4,4 difluoro-4-bora-3alpha,4alpha-diaza-s-indacene; ROX, 6-carboxy-X-rhodamine; R6G, 6-carboxyrhodamine-6G). Each nucleotide analogue consists of a different fluorophore attached to the 5 position of the pyrimidines and the 7 position of the purines through a photocleavable 2-nitrobenzyl linker. After verifying that these nucleotides could be successfully incorporated into a growing DNA strand in a solution-phase polymerase reaction and the fluorophore could be cleaved using laser irradiation ( approximately 355 nm) in 10 sec, we then performed an SBS reaction on a chip that contains a self-priming DNA template covalently immobilized by using 1,3-dipolar azide-alkyne cycloaddition. The DNA template was produced by PCR, using an azido-labeled primer, and the self-priming moiety was attached to the immobilized DNA template by enzymatic ligation. Each cycle of SBS consists of the incorporation of the photocleavable fluorescent nucleotide into the DNA, detection of the fluorescent signal, and photocleavage of the fluorophore. The entire process was repeated to identify 12 continuous bases in the DNA template. These results demonstrate that photocleavable fluorescent nucleotide analogues can be incorporated accurately into a growing DNA strand during a polymerase reaction in solution and on a chip. Moreover, all four fluorophores can be detected and then efficiently cleaved using near-UV irradiation, thereby allowing continuous identification of the DNA template sequence. Optimization of the steps involved in this SBS approach will further increase the read-length. PMID- 15829589 TI - Design and synthesis of a 3'-O-allyl photocleavable fluorescent nucleotide as a reversible terminator for DNA sequencing by synthesis. AB - DNA sequencing by synthesis (SBS) offers an approach for potential high throughput sequencing applications. In this method, the ability of an incoming nucleotide to act as a reversible terminator for a DNA polymerase reaction is an important requirement to unambiguously determine the identity of the incorporated nucleotide before the next nucleotide is added. A free 3'-OH group on the terminal nucleotide of the primer is necessary for the DNA polymerase to incorporate an incoming nucleotide. Therefore, if the 3'-OH group of an incoming nucleotide is capped by a chemical moiety, it will cause the polymerase reaction to terminate after the nucleotide is incorporated into the DNA strand. If the capping group is subsequently removed to generate a free 3'-OH, the polymerase reaction will reinitialize. We report here the design and synthesis of a 3' modified photocleavable fluorescent nucleotide, 3'-O-allyl-dUTP-PC-Bodipy-FL-510 (PC-Bodipy, photocleavable 4,4-difluoro-4-bora-3alpha,4alpha-diaza-s-indacene), as a reversible terminator for SBS. This nucleotide analogue contains an allyl moiety capping the 3'-OH group and a fluorophore Bodipy-FL-510 linked to the 5 position of the uracil through a photocleavable 2-nitrobenzyl linker. Here, we have shown that this nucleotide is a good substrate for a DNA polymerase. After the nucleotide was successfully incorporated into a growing DNA strand and the fluorophore was photocleaved, the allyl group was removed by using a Pd-catalyzed reaction to reinitiate the polymerase reaction, thereby establishing the feasibility of using such nucleotide analogues as reversible terminators for SBS. PMID- 15829590 TI - Activation of spinal d1/d5 receptors induces late-phase LTP of C-fiber-evoked field potentials in rat spinal dorsal horn. AB - Long-term potentiation (LTP) of C-fiber-evoked field potentials in spinal dorsal horn may be relevant to pathological pain. Our previous work has shown that the late phase of the spinal LTP is protein synthesis-dependent. Considerable evidence has accumulated that dopamine D1/D5 receptors are important for late phase LTP in hippocampus. In this study, the role of D1/D5 receptors in LTP of C fiber-evoked field potentials in spinal dorsal horn was evaluated in urethan anesthetized Sprague-Dawley rats. We found the following. 1) Spinal application of SKF 38393, a D1/D5 receptor agonist, induced a slowly developed LTP of C-fiber evoked field potentials, lasting for >10 h, and the effect was blocked by the D1/D5 antagonist SCH 23390, whereas a D2 receptor agonist (quinpirole) induced depression of C-fiber responses, lasting for 2 h. 2) The potentiation produced by D1/D5 receptor agonist occluded the late phase but not the early phase of the spinal LTP produced by tetanic stimulation. 3) SCH 23390 selectively depressed the late-phase LTP, when applied 40 min before tetanic stimulation. 4) The D1/D5 agonist-induced potentiation is blocked by the protein synthesis inhibitor anisomycin. 5) Activation of protein kinase A by spinal application of 8-Br-cAMP also induced spinal LTP, and the action occluded the potentiation induced by the D1/D5 receptor agonist. These results suggest that the spinal D1/D5 receptors participate in the protein synthesis-dependent late-phase LTP of C-fiber-evoked field potentials in spinal dorsal horn through the cAMP signaling pathway. PMID- 15829591 TI - In vivo analysis of proprioceptive coding and its antidromic modulation in the freely behaving crayfish. AB - Although sensory nerves in vitro are known to convey both orthodromic (sensory) and antidromic (putatively modulating) action potentials, in most cases very little is known about their bidirectional characteristics in intact animals. Here, we have investigated both the sensory coding properties and antidromic discharges that occur during real walking in the freely behaving crayfish. The activity of the sensory nerve innervating the proprioceptor CBCO, a chordotonal organ that monitors both angular movement and position of the coxo-basipodite (CB) joint, which is implicated in vertical leg movements, was recorded chronically along with the electromyographic activity of the muscles that control CB joint movements. Two wire electrodes placed on the sensory nerve were used to discriminate orthodromic from antidromic action potentials and thus allowed for analysis of both sensory coding and antidromic discharges. A distinction is proposed between 3 main classes of sensory neuron, according to their firing in relation to levator muscle activity during free walking. In parallel, we describe 2 types of antidromic activity: one produced exclusively during motor activity and a second produced both during and in the absence of motor activity. A negative correlation was found between the activity of sensory neurons in each of the 3 classes and identified antidromic discharges during walking. Finally, a state-dependent plasticity of CBCO nerve activity has been found by which the distribution of sensory orthodromic and antidromic activity changes with the physiological state of the biomechanical apparatus. PMID- 15829592 TI - Development of sound localization mechanisms in the mongolian gerbil is shaped by early acoustic experience. AB - Sound localization is one of the most important tasks performed by the auditory system. Differences in the arrival time of sound at the two ears are the main cue to localize low-frequency sound in the azimuth. In the mammalian brain, such interaural time differences (ITDs) are encoded in the auditory brain stem; first by the medial superior olive (MSO) and then transferred to higher centers, such as the dorsal nucleus of the lateral lemniscus (DNLL), a brain stem nucleus that gets a direct input from the MSO. Here we demonstrate for the first time that ITD sensitivity in gerbils undergoes a developmental maturation after hearing onset. We further show that this development can be disrupted by altering the animal's acoustic experience during a critical period. In animals that had been exposed to omnidirectional white noise during a restricted time period right after hearing onset, ITD tuning did not develop normally. Instead, it was similar to that of juvenile animals 3 days after hearing onset, with the ITD functions not adjusted to the physiological range. Animals that had been exposed to omnidirectional noise as adults did not show equivalent abnormal ITD tuning. The development presented here is in contrast to that of the development of neuronal representation of ITDs in the midbrain of barn owls and interaural intensity differences in ferrets, where the representations are adjusted by an interaction of auditory and visual inputs. The development of ITD tuning presented here most likely depends on normal acoustic experience and may be related to the maturation of inhibitory inputs to the ITD detector itself. PMID- 15829593 TI - Dynamic model of the octopus arm. II. Control of reaching movements. AB - The dynamic model of the octopus arm described in the first paper of this 2-part series was used here to investigate the neural strategies used for controlling the reaching movements of the octopus arm. These are stereotypical extension movements used to reach toward an object. In the dynamic model, sending a simple propagating neural activation signal to contract all muscles along the arm produced an arm extension with kinematic properties similar to those of natural movements. Control of only 2 parameters fully specified the extension movement: the amplitude of the activation signal (leading to the generation of muscle force) and the activation traveling time (the time the activation wave takes to travel along the arm). We found that the same kinematics could be achieved by applying activation signals with different activation amplitudes all exceeding some minimal level. This suggests that the octopus arm could use minimal amplitudes of activation to generate the minimal muscle forces required for the production of the desired kinematics. Larger-amplitude signals would generate larger forces that increase the arm's stability against perturbations without changing the kinematic characteristics. The robustness of this phenomenon was demonstrated by examining activation signals with either a constant or a bell shaped velocity profile. Our modeling suggests that the octopus arm biomechanics may allow independent control of kinematics and resistance to perturbation during arm extension movements. PMID- 15829594 TI - Dynamic model of the octopus arm. I. Biomechanics of the octopus reaching movement. AB - The octopus arm requires special motor control schemes because it consists almost entirely of muscles and lacks a rigid skeletal support. Here we present a 2D dynamic model of the octopus arm to explore possible strategies of movement control in this muscular hydrostat. The arm is modeled as a multisegment structure, each segment containing longitudinal and transverse muscles and maintaining a constant volume, a prominent feature of muscular hydrostats. The input to the model is the degree of activation of each of its muscles. The model includes the external forces of gravity, buoyancy, and water drag forces (experimentally estimated here). It also includes the internal forces generated by the arm muscles and the forces responsible for maintaining a constant volume. Using this dynamic model to investigate the octopus reaching movement and to explore the mechanisms of bend propagation that characterize this movement, we found the following. 1) A simple command producing a wave of muscle activation moving at a constant velocity is sufficient to replicate the natural reaching movements with similar kinematic features. 2) The biomechanical mechanism that produces the reaching movement is a stiffening wave of muscle contraction that pushes a bend forward along the arm. 3) The perpendicular drag coefficient for an octopus arm is nearly 50 times larger than the tangential drag coefficient. During a reaching movement, only a small portion of the arm is oriented perpendicular to the direction of movement, thus minimizing the drag force. PMID- 15829595 TI - Phase-response curves give the responses of neurons to transient inputs. AB - Neuronal firing is determined largely by incoming barrages of excitatory postsynaptic potentials (EPSPs), each of which produce a transient increase in firing probability. To measure the effects of weak transient inputs on firing probability of cortical neurons, we compute phase-response curves (PRCs). PRCs, whose shape can be related to the dynamics of spike generation, document the changes in timing of spikes caused by an EPSP in a repetitively firing neuron as a function of when it arrives in the interspike interval (ISI). The PRC can be exactly related to the poststimulus time histogram (PSTH) so that knowledge of one uniquely determines the other. Typically, PRCs have zero values at the start and end of the ISI, where EPSPs have minimal effects and a peak in the middle. Where the peak occurs depends in part on the firing properties of neurons. The PRC can have regions of positivity and negativity corresponding respectively to speeding up and slowing down the time of the next spike. A simple canonical model for spike generation is introduced that shows how both the background firing rate and the degree of postspike afterhyperpolarization contribute to the shape of the PRC and thus to the PSTH. PRCs in strongly adapting neurons are highly skewed to the right (indicating a higher change in probability when the EPSPs appear late in the ISI) and can have negative regions (indicating a decrease in firing probability) early in the ISI. The PRC becomes more skewed to the right as the firing rate decreases. Thus at low firing rates, the spikes are triggered preferentially by inputs that occur only during a small time interval late in the ISI. This implies that the neuron is more of a coincidence detector at low firing frequencies and more of an integrator at high frequencies. The steady-state theory is shown to also hold for slowly varying inputs. PMID- 15829596 TI - Monoaminergic establishment of rostrocaudal gradients of rhythmicity in the neonatal mouse spinal cord. AB - Bath application of monoamines is a potent method for evoking locomotor activity in neonatal rats and mice. Monoamines also promote functional recovery in adult animals with spinal cord injuries by activating spinal cord networks. However, the mechanisms of their actions on spinal networks are largely unknown. In this study, we tested the hypothesis that monoamines establish rostrocaudal gradients of rhythmicity in the thoracolumbar spinal cord. Isolated neonatal mouse spinal cord preparations (P0-P2) were used. To assay excitability of networks by monoamines, we evoked a disinhibited rhythm by bath application of picrotoxin and strychnine and recorded neurograms from several thoracolumbar ventral roots. We first established that rostral and caudal segments of the thoracolumbar spinal cord had equal excitability by completely transecting preparations at the L3 segmental level and recording the frequency of the disinhibited rhythm from both segments. Next we established that a majority of ventral interneurons retrogradely labeled by calcium green dextran were active during network activity. We then bath applied combinations of monoaminergic agonists [5-HT and dopamine (DA)] known to elicit locomotor activity. Our results show that monoamines establish rostrocaudal gradients of rhythmicity in the thoracolumbar spinal cord. This may be one mechanism by which combinations of monoaminergic compounds normally stably activate locomotor networks. PMID- 15829597 TI - Motor neuron firing dysfunction in spastic patients with primary lateral sclerosis. AB - Patients with corticospinal tract dysfunction have slow voluntary movements with brisk stretch reflexes and spasticity. Previous studies reported reduced firing rates of motor units during voluntary contraction. To assess whether this firing behavior occurs because motor neurons do not respond normally to excitatory inputs, we studied motor units in patients with primary lateral sclerosis, a degenerative syndrome of progressive spasticity. Firing rates were measured from motor units in the wrist extensor muscles at varying levels of voluntary contraction < or =10% maximal force. At each force level, the firing rate was measured with and without added muscle vibration, a maneuver that repetitively activates muscle spindles. In motor units from age-matched control subjects, the firing rate increased with successively stronger contractions as well as with the addition of vibration at each force level. In patients with primary lateral sclerosis, motor-unit firing rates remained stable, or in some cases declined, with progressively stronger contractions or with muscle vibration. We conclude that excitatory inputs produce a blunted response in motor neurons in patients with primary lateral sclerosis compared with age-matched controls. The potential explanations include abnormal activation of voltage-activated channels that produce stable membrane plateaus at low voltages, abnormal recruitment of the motor pool, or tonic inhibition of motor neurons. PMID- 15829598 TI - On the potential role of the corticospinal tract in the control and progressive adaptation of the soleus h-reflex during backward walking. AB - When untrained subjects walk backward on a treadmill, an unexpectedly large amplitude soleus H-reflex occurs in the midswing phase of backward walking. We hypothesized that activity in the corticospinal tract (CST) during midswing depolarizes the soleus alpha-motoneurons subliminally and thus brings them closer to threshold. To test this hypothesis, transcranial magnetic stimulation (TMS) was applied to the leg area of the motor cortex (MCx) during backward walking. Motor-evoked potentials (MEPs) were recorded from the soleus and tibialis anterior (TA) muscles in untrained subjects at different phases of the backward walking cycle. We reasoned that if soleus MEPs could be elicited in midswing, while the soleus is inactive, this would be strong evidence for increased postsynaptic excitability of the alpha-motoneurons. In the event, we found that in untrained subjects, despite the presence of an unexpectedly large H-reflex in midswing, no soleus MEPs were observed at that time. The soleus MEPs were in phase with the soleus electromyographic (EMG) activity during backward walking. Soleus MEPs increased more rapidly as a function of the EMG activity during voluntary activity than during backward walking. Furthermore, a conditioning stimulus to the motor cortex facilitated the soleus H-reflex at rest and during voluntary plantarflexion but not in the midswing phase of backward walking. With daily training at walking backward, the time at which the H-reflex began to increase was progressively delayed until it coincided with the onset of soleus EMG activity, and its amplitude was considerably reduced compared with its value on the first experimental day. By contrast, no changes were observed in the timing or amplitude of soleus MEPs with training. Taken together, these observations make it unlikely that the motor cortex via the CST is involved in control of the H-reflex during the backward step cycle of untrained subjects nor in its progressive adaptation with training. Our observations raise the possibility that the large amplitude of H-reflex in untrained subjects and its adaptation with training are mainly due to control of presynaptic inhibition of Ia-afferents by other descending tracts. PMID- 15829599 TI - GABAergic modulation of the activity of globus pallidus neurons in primates: in vivo analysis of the functions of GABA receptors and GABA transporters. AB - Neurons in the external and internal segment of the globus pallidus (GPe and GPi, respectively) receive substantial GABAergic inputs from the striatum and through axon collaterals of neighboring pallidal neurons. The effects of GABA on pallidal activity depend on the synaptic localization of GABA receptors and the distribution and activity of GABA transporters (GATs). To explore the contribution of GABA receptors and transporters to pallidal function, we recorded the activity of single neurons in GPe or GPi before, during, and after local microinjections of GABAergic compounds in awake rhesus monkeys. Activation of GABA(A) or GABA(B) receptors with muscimol or baclofen, respectively, inhibited pallidal activity. These effects were reversed by concomitant infusion of the respective GABA receptor antagonists, gabazine and CGP-55845. Given alone, the antagonists were without consistent effect. Application of the selective GAT-1 inhibitor, SKF-89976A, and the semiselective GAT-3 blocker, SNAP-5114, decreased pallidal activity. Both GAT inhibitors increased GABA levels in the pallidum, as measured by microdialysis. Electron microscopic observations revealed that these transporters are located on glial processes and unmyelinated axonal segments, but rarely on terminals. Our results indicate that activation of GABA(A) and GABA(B) receptors inhibits neuronal activity in both segments of the pallidum. GAT-1 and GAT-3 are involved in the modulation of endogenous GABA levels and may be important in regulating the extrasynaptic levels of GABA. Together with previous evidence that a considerable proportion of pallidal GABA receptors are located outside the synaptic cleft, our experiments strongly support the importance of extrasynaptic GABAergic transmission in the primate pallidum. PMID- 15829600 TI - MicroRNA-directed regulation of Arabidopsis AUXIN RESPONSE FACTOR17 is essential for proper development and modulates expression of early auxin response genes. AB - The phytohormone auxin plays critical roles during plant growth, many of which are mediated by the auxin response transcription factor (ARF) family. MicroRNAs (miRNAs), endogenous 21-nucleotide riboregulators, target several mRNAs implicated in auxin responses. miR160 targets ARF10, ARF16, and ARF17, three of the 23 Arabidopsis thaliana ARF genes. Here, we describe roles of miR160-directed ARF17 posttranscriptional regulation. Plants expressing a miRNA-resistant version of ARF17 have increased ARF17 mRNA levels and altered accumulation of auxin inducible GH3-like mRNAs, YDK1/GH3.2, GH3.3, GH3.5, and DFL1/GH3.6, which encode auxin-conjugating proteins. These expression changes correlate with dramatic developmental defects, including embryo and emerging leaf symmetry anomalies, leaf shape defects, premature inflorescence development, altered phyllotaxy along the stem, reduced petal size, abnormal stamens, sterility, and root growth defects. These defects demonstrate the importance of miR160-directed ARF17 regulation and implicate ARF17 as a regulator of GH3-like early auxin response genes. Many of these defects resemble phenotypes previously observed in plants expressing viral suppressors of RNA silencing and plants with mutations in genes important for miRNA biogenesis or function, providing a molecular rationale for phenotypes previously associated with more general disruptions of miRNA function. PMID- 15829601 TI - Auxin and light control of adventitious rooting in Arabidopsis require ARGONAUTE1. AB - Adventitious rooting is a quantitative genetic trait regulated by both environmental and endogenous factors. To better understand the physiological and molecular basis of adventitious rooting, we took advantage of two classes of Arabidopsis thaliana mutants altered in adventitious root formation: the superroot mutants, which spontaneously make adventitious roots, and the argonaute1 (ago1) mutants, which unlike superroot are barely able to form adventitious roots. The defect in adventitious rooting observed in ago1 correlated with light hypersensitivity and the deregulation of auxin homeostasis specifically in the apical part of the seedlings. In particular, a clear reduction in endogenous levels of free indoleacetic acid (IAA) and IAA conjugates was shown. This was correlated with a downregulation of the expression of several auxin-inducible GH3 genes in the hypocotyl of the ago1-3 mutant. We also found that the Auxin Response Factor17 (ARF17) gene, a potential repressor of auxin inducible genes, was overexpressed in ago1-3 hypocotyls. The characterization of an ARF17-overexpressing line showed that it produced fewer adventitious roots than the wild type and retained a lower expression of GH3 genes. Thus, we suggest that ARF17 negatively regulates adventitious root formation in ago1 mutants by repressing GH3 genes and therefore perturbing auxin homeostasis in a light dependent manner. These results suggest that ARF17 could be a major regulator of adventitious rooting in Arabidopsis. PMID- 15829602 TI - Crown rootless1, which is essential for crown root formation in rice, is a target of an AUXIN RESPONSE FACTOR in auxin signaling. AB - Although the importance of auxin in root development is well known, the molecular mechanisms involved are still unknown. We characterized a rice (Oryza sativa) mutant defective in crown root formation, crown rootless1 (crl1). The crl1 mutant showed additional auxin-related abnormal phenotypic traits in the roots, such as decreased lateral root number, auxin insensitivity in lateral root formation, and impaired root gravitropism, whereas no abnormal phenotypic traits were observed in aboveground organs. Expression of Crl1, which encodes a member of the plant specific ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES protein family, was localized in tissues where crown and lateral roots are initiated and overlapped with beta-glucuronidase staining controlled by the DR5 promoter. Exogenous auxin treatment induced Crl1 expression without de novo protein biosynthesis, and this induction required the degradation of AUXIN/INDOLE-3-ACETIC ACID proteins. Crl1 contains two putative auxin response elements (AuxREs) in its promoter region. The proximal AuxRE specifically interacted with a rice AUXIN RESPONSE FACTOR (ARF) and acted as a cis-motif for Crl1 expression. We conclude that Crl1 encodes a positive regulator for crown and lateral root formation and that its expression is directly regulated by an ARF in the auxin signaling pathway. PMID- 15829604 TI - Arabidopsis tic110 is essential for the assembly and function of the protein import machinery of plastids. AB - The translocon at the inner envelope membrane of chloroplasts (Tic) plays a central role in plastid biogenesis by coordinating the sorting of nucleus-encoded preproteins across the inner membrane and coordinating the interactions of preproteins with the processing and folding machineries of the stroma. Despite these activities, the precise roles of known Tic proteins in translocation, sorting, and preprotein maturation have not been defined. In this report, we examine the in vivo function of a major Tic component, Tic110. We demonstrate that Arabidopsis thaliana Tic110 (atTic110) is essential for plastid biogenesis and plant viability. The downregulation of atTic110 expression results in the reduced accumulation of a wide variety of plastid proteins. The expression of dominant negative mutants of atTic110 disrupts assembly of Tic complexes and the translocation of preproteins across the inner envelope membrane. Together, these data suggest that Tic110 plays a general role in the import of nuclear-encoded preproteins as a common component of Tic complexes. PMID- 15829603 TI - MicroRNA directs mRNA cleavage of the transcription factor NAC1 to downregulate auxin signals for arabidopsis lateral root development. AB - Although several plant microRNAs (miRNAs) have been shown to play a role in plant development, no phenotype has yet been associated with a reduction or loss of expression of any plant miRNA. Arabidopsis thaliana miR164 was predicted to target five NAM/ATAF/CUC (NAC) domain-encoding mRNAs, including NAC1, which transduces auxin signals for lateral root emergence. Here, we show that miR164 guides the cleavage of endogenous and transgenic NAC1 mRNA, producing 3'-specific fragments. Cleavage was blocked by NAC1 mutations that disrupt base pairing with miR164. Compared with wild-type plants, Arabidopsis mir164a and mir164b mutant plants expressed less miR164 and more NAC1 mRNA and produced more lateral roots. These mutant phenotypes can be complemented by expression of the appropriate MIR164a and MIR164b genomic sequences. By contrast, inducible expression of miR164 in wild-type plants led to decreased NAC1 mRNA levels and reduced lateral root emergence. Auxin induction of miR164 was mirrored by an increase in the NAC1 mRNA 3' fragment, which was not observed in the auxin-insensitive mutants auxin resistant1 (axr1-12), axr2-1, and transport inhibitor response1. Moreover, the cleavage-resistant form of NAC1 mRNA was unaffected by auxin treatment. Our results indicate that auxin induction of miR164 provides a homeostatic mechanism to clear NAC1 mRNA to downregulate auxin signals. PMID- 15829605 TI - Coordination of nuclear and mitochondrial genome expression during mitochondrial biogenesis in Arabidopsis. AB - Mitochondrial biogenesis and function require the regulated and coordinated expression of nuclear and mitochondrial genomes throughout plant development and in response to cellular and environmental signals. To investigate the levels at which the expression of nuclear and mitochondrially encoded proteins is coordinated, we established an Arabidopsis thaliana cell culture system to modulate mitochondrial biogenesis in response to sugar starvation and refeeding. Sucrose deprivation led to structural changes in mitochondria, a decrease in mitochondrial volume, and a reduction in the rate of cellular respiration. All these changes could be reversed by the readdition of sucrose. Analysis of the relative mRNA transcript abundance of genes encoding nuclear and mitochondrially encoded proteins revealed that there was no coordination of expression of the two genomes at the transcript level. An analysis of changes in abundance and assembly of nuclear-encoded and mitochondrially encoded subunits of complexes I to V of the mitochondrial inner membrane in organello protein synthesis and competence for protein import by isolated mitochondria suggested that coordination occurs at the level of protein-complex assembly. These results further suggest that expression of the mitochondrial genome is insensitive to the stress imposed by sugar starvation and that mitochondrial biogenesis is regulated by changes in nuclear gene expression and coordinated at the posttranslational level. PMID- 15829607 TI - Structural and kinetic basis for substrate selectivity in Populus tremuloides sinapyl alcohol dehydrogenase. AB - We describe the three-dimensional structure of sinapyl alcohol dehydrogenase (SAD) from Populus tremuloides (aspen), a member of the NADP(H)-dependent dehydrogenase family that catalyzes the last reductive step in the formation of monolignols. The active site topology revealed by the crystal structure substantiates kinetic results indicating that SAD maintains highest specificity for the substrate sinapaldehyde. We also report substantial substrate inhibition kinetics for the SAD-catalyzed reduction of hydroxycinnamaldehydes. Although SAD and classical cinnamyl alcohol dehydrogenases (CADs) catalyze the same reaction and share some sequence identity, the active site topology of SAD is strikingly different from that predicted for classical CADs. Kinetic analyses of wild-type SAD and several active site mutants demonstrate the complexity of defining determinants of substrate specificity in these enzymes. These results, along with a phylogenetic analysis, support the inclusion of SAD in a plant alcohol dehydrogenase subfamily that includes cinnamaldehyde and benzaldehyde dehydrogenases. We used the SAD three-dimensional structure to model several of these SAD-like enzymes, and although their active site topologies largely mirror that of SAD, we describe a correlation between substrate specificity and amino acid substitution patterns in their active sites. The SAD structure thus provides a framework for understanding substrate specificity in this family of enzymes and for engineering new enzyme specificities. PMID- 15829606 TI - Disruptions of the Arabidopsis Enoyl-CoA reductase gene reveal an essential role for very-long-chain fatty acid synthesis in cell expansion during plant morphogenesis. AB - In the absence of cell migration, plant architecture is largely determined by the direction and extent of cell expansion during development. In this report, we show that very-long-chain fatty acid (VLCFA) synthesis plays an essential role in cell expansion. The Arabidopsis thaliana eceriferum10 (cer10) mutants exhibit severe morphological abnormalities and reduced size of aerial organs. These mutants are disrupted in the At3g55360 gene, previously identified as a gene coding for enoyl-CoA reductase (ECR), an enzyme required for VLCFA synthesis. The absence of ECR activity results in a reduction of cuticular wax load and affects VLCFA composition of seed triacylglycerols and sphingolipids, demonstrating in planta that ECR is involved in all VLCFA elongation reactions in Arabidopsis. Epidermal and seed-specific silencing of ECR activity resulted in a reduction of cuticular wax load and the VLCFA content of seed triacylglycerols, respectively, with no effects on plant morphogenesis, suggesting that the developmental phenotypes arise from abnormal sphingolipid composition. Cellular analysis revealed aberrant endocytic membrane traffic and defective cell expansion underlying the morphological defects of cer10 mutants. PMID- 15829608 TI - The influence of chemical gustatory stimuli and oral anaesthesia on healthy human pharyngeal swallowing. AB - This study explored the effects of taste and oral anaesthesia on human sequential swallowing. Subjects were healthy adults (n = 42, mean age 28 years, 21 females), investigated by means of a water swallow test. Taste stimuli comprised quinine, glucose, citrus and saline solutions compared with neutral water. Oral anaesthesia comprised topical lidocaine at doses of 10, 20 and 40 mg and compared with placebo. Data were collected on swallowing speed (volume per second), inter swallow interval and swallowing capacity (volume per swallow). Compared with water, glucose, citrus and saline reduced swallowing speed (10.94 +/- 0.89 versus 9.56 +/- 0.79, 9.33 +/- 1.19, 9.37 +/- 0.92 ml/s respectively, P < 0.05). Inter swallow interval was increased only by quinine and saline (1.47 +/- 1.11 versus 2.13 +/- 0.34 and 1.92 +/- 0.31 s, P < 0.04). Swallowing capacity was only marginally increased by quinine (P = 0.0759). Compared with the placebo, only 40 mg of lidocaine altered swallowing, immediately reducing the swallowing speed (7.89 +/- 2.34 versus 10.11 +/- 3.26 ml/s, P < 0.05) and increasing inter-swallow interval (1.67 +/- 0.38 versus 1.45 +/- 0.29 s, P < 0.01) without affecting capacity. By 15 min all measures except sensory thresholds had returned to baseline values. Thus, swallowing function is highly influenced by chemosensory input, providing insight into how oral sensation regulates pharyngeal swallowing. PMID- 15829609 TI - Neuronal representations of stimuli in the mouth: the primate insular taste cortex, orbitofrontal cortex and amygdala. AB - The responses of 3687 neurons in the macaque primary taste cortex in the insula/frontal operculum, orbitofrontal cortex (OFC) and amygdala to oral sensory stimuli reveals principles of representation in these areas. Information about the taste, texture of what is in the mouth (viscosity, fat texture and grittiness, which reflect somatosensory inputs), temperature and capsaicin is represented in all three areas. In the primary taste cortex, taste and viscosity are more likely to activate different neurons, with more convergence onto single neurons particularly in the OFC and amygdala. The different responses of different OFC neurons to different combinations of these oral sensory stimuli potentially provides a basis for different behavioral responses. Consistently, the mean correlations between the representations of the different stimuli provided by the population of OFC neurons were lower (0.71) than for the insula (0.81) and amygdala (0.89). Further, the encoding was more sparse in the OFC (0.67) than in the insula (0.74) and amygdala (0.79). The insular neurons did not respond to olfactory and visual stimuli, with convergence occurring in the OFC and amygdala. Human psychophysics showed that the sensory spaces revealed by multidimensional scaling were similar to those provided by the neurons. PMID- 15829610 TI - Case study: using a virtual reality computer game to teach fire safety skills to children diagnosed with fetal alcohol syndrome. AB - OBJECTIVE: To assess the effectiveness of a computer-based virtual reality (VR) game in teaching five children diagnosed with fetal alcohol syndrome (FAS) fire safety skills and to generalize these skills to a real world simulation. METHOD: Children participated in a study by using a multiple baseline, multiple probe design. Before the game, no child could correctly describe what actions to take during a home fire. A computerized game allowed them to learn the recommended safety steps in a virtual world. Skill learning and real-world generalization were tested immediately after the intervention and at 1-week post-test. RESULTS: All children reached 100% accuracy on the computer intervention, defined as successfully completing each of the safety steps. At the 1-week follow-up, all the children were able to perform the steps correctly in a real world simulation. CONCLUSIONS: The results suggest that this method of intervention warrants further study as an educational delivery system for children with FAS. PMID- 15829611 TI - Prenatal alcohol exposure and ability, academic achievement, and school functioning in adolescence: a longitudinal follow-up. AB - BACKGROUND: Prenatal alcohol exposure is associated with learning, behavioral, and academic problems even in children without the fetal alcohol syndrome (FAS). OBJECTIVE: To examine the prenatal alcohol exposure and ability, academic achievement, and school functioning in adolescence. METHODS: In a longitudinal cohort, intelligence, academic performance, and school functioning were evaluated in 265 low socioeconomic status (SES) adolescents (M age = 15.1 years), 128 prenatally exposed to alcohol, 53 controls, and 84 special education students by using the Wechsler Intelligence Scale for Children, 3rd edition (WISC-III) and the Wechsler Individual Achievement Test (WIAT). School records were abstracted for grade point averages (GPA), standardized achievement test scores, conduct, attendance, and special education placement. RESULTS: Alcohol-affected youth had significantly lower IQs than those in the other three groups. CONCLUSION: Although academic achievement (WIAT scores) was most impaired in the special education group who showed lower performance over all as well as in reading and spelling, alcohol-affected youth showed significant deficits on mathematics subtests. There was no increased incidence of conduct problems in school records related to alcohol exposure. PMID- 15829612 TI - Advancing the science of perceptual accuracy in pediatric asthma and diabetes. AB - OBJECTIVES: To review research on perceptual accuracy in pediatric asthma and diabetes and to provide recommendations for future research efforts and clinical applications of the construct in these populations. METHODS: A literature search was conducted using Medline and PsychInfo databases as well as the bibliographies of relevant articles. RESULTS: Children and adolescents with asthma or diabetes evidence considerable variability in perceptual accuracy and frequently make clinically relevant errors that have the potential to affect self-management behavior. CONCLUSIONS: Recommendations for future research include studying distinct types of perceptual errors, empirically supporting the relationship between perceptual accuracy and relevant outcomes, identifying factors related to perceptual inaccuracy, and conducting longitudinal research and intervention studies. Recommendations for applying the construct in clinical practice include adopting an individualized approach to symptoms to guide patient education and management, identifying patients prone to making clinically relevant errors, and developing and implementing interventions to improve accuracy. PMID- 15829613 TI - Altered gene expression during rat Wolffian duct development following di(n butyl) phthalate exposure. AB - Di(n-butyl) phthalate (DBP) is a common plasticizer and solvent that disrupts androgen-dependent male reproductive development in rats. In utero exposure to 500 mg/kg/day DBP on gestation days (GD) 12 to 21 decreases androgen biosynthetic enzymes, resulting in decreased fetal testicular testosterone levels. One consequence of prenatal DBP exposure is malformed epididymides in adult rats. Reduced fetal testosterone levels may be responsible for the malformation, since testosterone is required for Wolffian duct stabilization and their development into epididymides. Currently, little is understood about the molecular mechanisms of Wolffian duct differentiation. The objective of this study was to identify changes in gene expression associated with altered morphology of the proximal Wolffian duct following in utero exposure to DBP. Pregnant Crl:CD(R) (SD) rats were gavaged with corn oil vehicle or 500 mg/kg/day DBP from GD 12 to GD 19 or 21. There were only small morphological differences between control and DBP exposed Wolffian ducts on GD 19. On GD 21, 89% of male fetuses in the DBP dose group showed marked underdevelopment of Wolffian ducts, characterized by decreased coiling. RNA was isolated from Wolffian ducts on GD 19 and 21. Together with empirical information, cDNA microarrays were used to help identify candidate genes that could be associated with the morphological changes observed on GD 21. These candidate genes were analyzed by real-time RT-PCR. Changes in mRNA expression were observed in genes within the insulin-like growth factor (IGF) pathway, the matrix metalloproteinase (MMP) family, the extracellular matrix, and in other developmentally conserved signaling pathways. On GD 19, immunolocalization of IGF-1 receptor protein demonstrated an increase in cytoplasmic expression in the mesenchymal and epithelial cells. There was also a variable decrease in androgen receptor protein in ductal epithelial cells on GD 19. This study provides insight into the effects of antiandrogens on the molecular mechanisms involved in Wolffian duct development. The altered morphology and changes in gene expression following DBP exposure are suggestive of altered paracrine interactions between ductal epithelial cells and the surrounding mesenchyme during Wolffian duct differentiation due to lowered testosterone production. PMID- 15829614 TI - Genotoxicity of 4-hydroxy-2-nonenal in human colon tumor cells is associated with cellular levels of glutathione and the modulation of glutathione S-transferase A4 expression by butyrate. AB - The cellular production of 4-hydroxy-2-nonenal (HNE), a product of endogenous lipid peroxidation, constitutes a genotoxic risk factor for carcinogenesis. Our previous studies have shown that human HT29 colon cells developed resistance toward HNE injury after treatment with butyrate, a diet-associated gut fermentation product. This resistance was attributed to the induction of certain glutathione S-transferases (hGSTP1-1, hGSTM2-2, and hGSTA1-1) and also for the tripeptide glutathione (GSH) synthesizing enzymes. In the present study, we have investigated in HT29 cells whether hGSTA4-4, which has a high substrate specificity for HNE, was also inducible by butyrate and, thus, could contribute to the previously observed chemoresistance. In addition, we investigated if cellular depletion of GSH by L-buthionine-S,R-sulfoximine (BSO) enhances chemosensitivity to HNE injury in HT29 cells. Incubation of HT29 cells with butyrate (2-4 mM) significantly elicited a 1.8 to 3-fold upregulation of steady state hGSTA4 mRNA over 8-24 h after treatment. Moreover, 4 mM butyrate tended to increase hGSTA4-4 protein concentrations. Incubation with 100 microM BSO decreased cellular GSH levels by 77% without significant changes in cell viability. Associated with this was a 2-fold higher level of HNE-induced DNA damage as measured by the comet assay. Collectively, the results of this study and our previous work indicate that the genotoxicity of HNE is highly dependent on cellular GSH status and those GSTs that contribute toward HNE conjugation, including hGSTA4-4. Since HNE contributes to colon carcinogenesis, the favorable modulation of the GSH/GST system by butyrate may contribute to chemoprevention and reduction of the risks. PMID- 15829615 TI - Atrazine and the hypothalamo-pituitary-gonadal axis in sexually maturing precocial birds: studies in male Japanese quail. AB - The herbicide atrazine is a putative endocrine disruptor. The present studies investigated the effects of atrazine in male Japanese quail during sexual maturation. Atrazine was administered for two weeks in the diet or systemically to birds under long photoperiods. Atrazine had no effect on mortality but depressed both feed intake and growth (average daily gain [ADG] in g/day) at dietary concentrations of 1000 ppm. Atrazine in the diet at 10 ppm, but at no other concentrations, increased testes weight and gonadal-somatic-index and decreased the seminiferous tubule diameter-to-testis weight ratio. However, there were no effects on absolute tubule diameter, relative stage of testicular development, or the presence of a lumen. Atrazine in the diet at 1000 ppm increased circulating concentrations of testosterone but this effect was not observed consistently in all studies. Dietary atrazine at 10 ppm increased circulating concentrations of estradiol. Moreover, in one study, atrazine at 1000 ppm in the diet decreased circulating concentrations of luteinizing hormone. Atrazine administered systemically exerted no effect on indices of growth or reproduction. Atrazine did not mimic the effects of either estradiol or tamoxifen in male quail; thus, atrazine did not exhibit overt estrogenic or anti-estrogenic activity. Conversely, atrazine augmented the effects of testosterone and estradiol on testis regression, presumably by increasing the negative-feedback effects of these sex-steroids on follicle stimulating hormone secretion. It is concluded that atrazine up to 1000 ppm in the diet may exert some effects on reproductive development in sexually maturing male birds, but these are inconsistent and modest. PMID- 15829616 TI - The threshold of toxicological concern concept in risk assessment. AB - The concept that "safe levels of exposure" for humans can be identified for individual chemicals is central to the risk assessment of compounds with known toxicological profiles. The Threshold of Toxicological Concern (TTC) is a concept that refers to the establishment of a level of exposure for all chemicals, whether or not there are chemical-specific toxicity data, below which there would be no appreciable risk to human health. The concept proposes that a low level of exposure with a negligible risk can be identified for many chemicals, including those of unknown toxicity, based on knowledge of their chemical structures. The present paper aims to describe the history of the TTC principle, its use to date, its potential future applications and the incorporation of the TTC principle in the Risk Assessment paradigm. PMID- 15829617 TI - Differential gene expression in human peripheral blood mononuclear cells induced by cigarette smoke and its constituents. AB - In current molecular epidemiology studies, a wide range of methods are used to monitor early biological effects after exposure to xenobiotic agents. Gene expression profiling is considered a promising tool that may provide more sensitive, mechanism-based biomarkers. As a first step toward obtaining information on the applicability of gene expression profiles as a biomarker for early biological effects of carcinogen exposure, we conducted in vitro studies on human peripheral blood mononuclear cells (PBMC). We used cigarette smoke condensate (CSC) and a selection of its genotoxic constituents as model agents, applying cDNA microarray technology to investigate modulated gene expression. In independent experiments using cells from several donors, quiescent PBMC were exposed for 18 h, followed by gene expression analyses on a microarray containing 600 toxicologically relevant genes. The search for candidate biomarker genes was binomial: first we looked for genes responding similarly to all agents; second, for agent-specific genes. Many genes were significantly deregulated by all compounds, but as the direction of deregulation frequently differed per agent, they are not useful as generic biomarkers. Cigarette smoke condensate modulated the expression of many more genes than any of its constituents, with the largest effect in SERPINB2. The affected genes are involved in immune or stress responses, but surprisingly no genes involved in DNA damage response were modulated, and only a few in DNA repair. In conclusion, several genes have been identified as potential biomarkers for population studies on early biological effects caused by cigarette smoke exposure, but no genes were identified that represent a generic biomarker. PMID- 15829618 TI - Effects of exogenous sphinganine, sphingosine, and sphingosine-1-phosphate on relaxation and contraction of porcine thoracic aortic and pulmonary arterial rings. AB - Fumonisin mycotoxicosis in pigs causes a decrease in mean aortic pressure, an increase in mean pulmonary arterial pressure, and increases in serum concentrations of sphinganine (3.2 microM) and sphingosine (1.4 microM). To determine a causal relationship between the hemodynamic changes and sphingolipid alterations, we examined the in vitro effects of sphinganine, sphingosine, and sphingosine-1-phosphate on porcine aortic and pulmonary arterial rings. Both sphinganine and sphingosine relaxed un-contracted and phenylephrine-contracted aortic rings at > or = 10 microM and > or = 1 microM, respectively. Sphingosine (> or = 10 microM) relaxed un-contracted and phenylephrine-contracted pulmonary arterial rings, whereas sphingosine-1-phosphate (10 microM) contracted pulmonary arterial rings. Sphingosine (3 microM) also impaired the contractile response of pulmonary artery rings to 60 mM KCl. The results suggested that the systemic hypotension caused by fumonisin is mediated, in part, by increases in serum sphinganine and sphingosine concentrations, and the pulmonary hypertension is mediated, in part, by increased sphingosine-1-phosphate concentrations. PMID- 15829619 TI - An alien promoter capture as a primary step of the evolution of testes-expressed repeats in the Drosophila melanogaster genome. AB - Fertility of Drosophila melanogaster males is impaired due to the disruption of the silencing of the X-linked, testis-expressed, repeated Stellate (Ste) genes. Ste silencing is mediated by symmetric transcription of the paralogous Y-linked repeats and exerted by an RNA interference (RNAi) mechanism. Here we present a scenario for the origin of the Ste genes and their suppressors. The primary intermediate of their evolution emerged as a result of the acquisition of a preformed alien, testis-specific promoter. This intermediate is identified as a chimeric gene containing coding region of an autosomal gene for testis-specific protein kinase CK2. The 5' region of the chimera has been acquired from a member of a family of testis-expressed X-linked genes of unknown function. We propose that the evolution and amplification of the novel chimeric gene have led to the overproduction of the regulatory CK2 subunit in testes. The evolution of the Y linked descendants of the primary intermediate resulted in the RNAi-mediated suppression of excessive expression of the X-linked paralogs. The newly detected "dead family" of cognate repeats on the Y chromosome has contributed to the evolution of Ste and its suppressors via gene conversion. Our results show that RNAi silencing, considered as a defense against viruses and transposable elements, may be involved in the evolution of eukaryotic genomes. PMID- 15829620 TI - Directed mutagenesis confirms the functional importance of positively selected sites in polygalacturonase inhibitor protein. AB - Polygalacturonase inhibitor proteins (PGIPs) protect plants against invasion by diverse microbial and invertebrate enemies that use polygalacturonase (PG) to breach the plant cell wall. Directed mutagenesis has identified specific natural mutations conferring novel defensive capability in green bean PGIP against a specific fungal PG. These same sites are identified as positively selected by phylogenetic codon-substitution models, demonstrating the utility of such models for connecting retrospective comparative analyses with contemporary, ecologically relevant variation. PMID- 15829621 TI - Regulation of circulating gonadotropins by the negative effects of ovarian hormones in mares. AB - The functional and temporal relationships between circulating gonadotropins and ovarian hormones in mares during Days 7-27 (ovulation = Day 0) was studied using control, follicle ablation, and ovariectomy groups (n = 6 mares/group). In the follicle-ablation group, all follicles > or = 6 mm were ablated on Day 7, and every 2 days thereafter, newly emerging follicles were also ablated. Estradiol concentrations decreased (P < 0.01) similarly in the controls and the follicle ablation group between Days 7 and 11 and by Day 15 began to increase in the controls and continued to decrease in the follicle-ablation group. Concentrations of progesterone were not affected by follicle ablation, but diameter of the corpus luteum was greater (P < 0.05) by Day 21 in the follicle-ablation group; these results indicated that the follicles were involved in morphologic luteolysis, but not in functional luteolysis. Concentrations of LH were higher (P < 0.05) on Days 15 and 16 in the follicle-ablation group than in the controls, indicating an initial negative effect of follicles on LH. Immunoreactive inhibin and estradiol decreased (P < 0.0001) and FSH and LH increased (P < 0.05) within 1 or 2 days after ovariectomy; these changes occurred more slowly in the follicle ablation group. The maximum value for an FSH surge in each control mare was below the lower 95% confidence limit in the ovariectomy group. Maximum concentration for the periovulatory LH surge in the controls was not different from the mean maximum LH concentrations in the ovariectomy group. Our interpretation is that the gonadotropin surges resulted from changes in the magnitude of the negative effects of ovarian hormones on the positive effects of extraovarian control. There was no indication of a positive ovarian effect on either FSH or LH. PMID- 15829622 TI - Role of glutathione in reproductive tract secretions on mouse preimplantation embryo development. AB - We investigated the hypothesis that glutathione (GSH) in reproductive tract secretions (RTS) protects the preimplantation embryo from endogenous reactive oxygen species and is important for normal development during the embryo's sensitive period when it is incapable of synthesizing GSH de novo. Mice were administered buthionine sulfoximine (BSO) to inhibit GSH synthesis and decrease GSH concentration in RTS. Embryos were then allowed to develop either in vivo or in vitro in the presence of RTS and the GSH concentration of the embryos was quantified by HPLC and embryonic development was recorded. GSH concentration in RTS did not differ over the phases of the estrous cycle, but there were significant decreases in GSH concentration on Day 2 of gestation and due to BSO treatment. Embryos allowed to develop in vivo and in vitro in RTS with decreased GSH concentration did not exhibit decreased development or GSH concentration. Oocytes exposed to BSO during maturation in vivo experienced a significant decrease in GSH concentration and an increase in percent of degenerate embryos when compared with control. These data suggest that most of the GSH in RTS does not play a critical role in normal preimplantation embryo development but that GSH stored in the oocyte during maturation has an important role in subsequent embryo development. Our studies do not exclude the possibility that GSH in RTS plays an important role in protection of the preimplantation embryo during exposure to some toxicants. PMID- 15829623 TI - Identification of downregulated messenger RNAs in bovine granulosa cells of dominant follicles following stimulation with human chorionic gonadotropin. AB - Molecular determinants and mechanisms involved in ovarian follicular growth, ovulation, and luteinization are not well understood. The objective of this study was to identify genes expressed in bovine granulosa cells (GC) of dominant follicles (DF) and downregulated after hCG-induced ovulation, using the suppression subtractive hybridization (SSH). GC were collected from DF at Day 5 of the estrous cycle and from ovulatory follicles (OF) obtained 23 h following injection of hCG. A subtracted cDNA library (DF-OF) was generated and screened using unsubtracted (DF, OF) and subtracted (DF-OF, OF-DF) cDNAs as complex (32)P probes. A total of 32 nonredundant cDNAs were identified: 23 cDNAs matched with sequences of known biological function and 9 cDNAs with complete or partial sequences of undefined biological function. Detection of genes known to be downregulated during the periovulatory period in the bovine species, such as CPD, CYP11A1, CYP19A1, FSHR, LRP8/ ApoER2, and SERPINE2, validated the physiological model and analytical techniques used. For a subset of genes, such as ARFGAP3, CYP11A1, CYP19A1, FSHR, FST, GJA1, IDH3, INHBA, LHCGR, LHCGR lacking exon 10, PRC1, PRG1, RPA2, SCD, and TRIB2, gene expression profiles were compared by virtual Northern blot or reverse transcriptase-polymerase chain reaction from follicles obtained at different developmental stages. Results confirmed a downregulation of the respective mRNAs in GC of OF compared with that of DF. We conclude that we have identified novel genes that are downregulated by hCG in bovine GC of DF during the periovulatory period, which may contribute to follicular growth, ovulation, and/or luteinization. PMID- 15829624 TI - Sufficient progesterone-priming prior to estradiol stimulation is required for optimal induction of the cervical prostaglandin system in pregnant sheep at 0.7 gestations. AB - The purposes of this study were to determine the separate and interactive functions of progesterone and estradiol in regulating the cervical prostaglandin (PG) system in pregnant sheep at 0.7 gestations. At 106-108 days of gestational age (dGA), ewes were treated with vehicle for 14 days (n = 5) or vehicle for 12 days followed by estradiol 5 mg twice a day, intramuscularly for 2 days (n = 5) or progesterone 100 mg, twice a day, intramuscularly for 14 days (n = 5) or progesterone 100 mg twice a day, intramuscularly for 10 days and then 2 days vehicle followed by estradiol 5 mg twice a day intramuscularly for 2 days (n = 5). At 121-123 dGA, cervical tissues were obtained under halothane anesthesia. Cervical RNA and protein were extracted and analyzed for prostaglandin endoperoxide synthase 2 (COX2), two PGE(2) receptors, PTGER2 and PTGER4, and estrogen receptor alpha (ESR1) by Northern and Western blot analysis. Immunocytochemistry and in situ hybridization were applied to localize cellular distribution of COX2, PTGER2, and PTGER4 in the cervix. Data were analyzed by ANOVA. COX2 and PTGER4 mRNAs and proteins were increased (P < 0.05) in ewes treated with combined estradiol and progesterone but not in ewes treated with estradiol or progesterone alone compared with controls. ESR1 mRNA was increased in ewes treated with progesterone and estradiol plus progesterone. In contrast, PTGER2 mRNA and protein remained the same after all treatments. COX2 mRNA and protein were localized only in cervical glandular epithelial cells, whereas PTGER2 and PTGER4 were localized in both cervical glandular epithelial and smooth muscle cells. In conclusion, these data suggest that additional progesterone priming at 0.7 gestations synergizes with estradiol to induce cervical COX2, PTGER4, and ESR1 and support our hypothesis that stimulation of the cervical PG system by estradiol is optimized by sufficient progesterone priming in the pregnant sheep cervix. PMID- 15829625 TI - Nonoxynol-9 induces apoptosis of endometrial explants by both caspase-dependent and -independent apoptotic pathways. AB - Contraceptive microbicides formulated as vaginal gels offer the possibility of women-controlled contraception and prevention of HIV infection. The effects of these gels on the upper reproductive tract are largely unknown. The purpose of this study was to determine whether nonoxynol-9 (N-9) induces apoptosis in human endometrium using endometrial explant as a model. Apoptosis was determined by gel electrophoresis for the detection of DNA fragmentation and by immunohistochemistry using the M30 CytoDEATH and anti-cleaved caspase-3 (CASP3) antibodies for the detection of caspase activity. The ability of the broad spectrum caspase inhibitor and CASP3-specific inhibitor to prevent N-9-induced cell death was measured. Expression of apoptosis-related genes such as BCL2, BAX, Fas receptor (FAS), and Fas ligand (FASLG) was quantified using real-time polymerase chain reaction (PCR) analysis. This study demonstrated that N-9 induced DNA fragmentation and caspase activity in endometrial explants in a dose- and time-dependent manner. Caspase inhibitors did not fully prevent the N-9 induced DNA fragmentation. Real-time PCR analysis revealed that FAS and FASLG were largely increased following N-9 treatment. Together, these results suggested that apoptosis triggered by N-9 in endometrial explants is mediated upstream via FAS and FASLG, followed by CASP3 activation leading to final cell death. It appears that other factors besides caspases are also involved in the N-9-induced apoptosis. PMID- 15829626 TI - c-Jun NH2-terminal kinase-interacting protein-3 facilitates phosphorylation and controls localization of amyloid-beta precursor protein. AB - Abnormal phosphorylation of amyloid-beta precursor protein (APP) is a pathologic feature of Alzheimer's disease. To begin to understand the mechanism of APP phosphorylation, we studied this process in differentiating neurons under normal physiological conditions. We found that c-Jun NH2-terminal kinase (JNK), not cyclin-dependent kinase 5, is required for APP phosphorylation, leading to localized accumulation of phosphorylated APP (pAPP) in neurites. We show that JNK interacting protein-3 (JIP-3), a JNK scaffolding protein that does not bind APP, selectively increases APP phosphorylation, accumulation of pAPP into processes, and stimulates process extension in both neurons and COS-1 cells. Downregulation of JIP-3 by small interfering RNA impairs neurite extension and reduces the amount of localized pAPP. Finally, whereas stress-activated JNK generates pAPP only in the cell body, concomitant expression of JIP-3 restores pAPP accumulation into neurites. Thus, APP phosphorylation, transport of the generated pAPP into neurites, and neurite extension are interdependent processes regulated by JIP 3/JNK, in a pathway distinct from stress-activated JNK signaling. PMID- 15829627 TI - AMPA receptor binding cleft mutations that alter affinity, efficacy, and recovery from desensitization. AB - Glutamate binds to AMPA receptors within a deep cleft between two globular protein domains (domains 1 and 2). Once glutamate binds, the cleft closes, and agonist-bound structures of the isolated ligand binding core suggest that closure of the binding cleft is sufficiently complete that it essentially prevents ligand dissociation. There is also considerable evidence supporting the view that cleft closure is the initial conformational change that triggers receptor activation and desensitization, and it has been clearly demonstrated that there is a correlation between the degree of cleft closure and agonist efficacy. It is unknown, however, whether the stability of binding cleft closure also influences receptor-channel properties. The crystallographic structures indicate that closed cleft conformations are stabilized by the formation of hydrogen bonds that involve amino acid side chains of residues in domains 1 and 2. We show here that mutations that disrupt one such cross-cleft hydrogen bond (in the AMPA receptor subunit GluR2) decrease both agonist affinity and efficacy. The same mutations also hasten recovery from desensitization. We conclude that the stability of binding cleft closure has a significant impact on AMPA receptor function and is a major determinant of the apparent affinity of agonists. The results suggest that the stability of cleft closure has been tuned so that glutamate dissociates as rapidly as possible yet remains a full agonist. PMID- 15829628 TI - Metabotropic glutamate receptors and dopamine receptors cooperate to enhance extracellular signal-regulated kinase phosphorylation in striatal neurons. AB - Striatal medium spiny neurons are an important site of convergence for signaling mediated by the neurotransmitters dopamine and glutamate. We report that in striatal neurons in primary culture, signaling through group I metabotropic glutamate receptors (mGluRs) 1/5 and the D1 class of dopamine receptors (DRs) 1/5 converges to increase phosphorylation of the mitogen-activated protein kinase ERK2 (extracellular signal-regulated kinase 2). Induction of mitogen-activated protein kinase kinase-dependent signaling cascades by either mGluR1/5 or DR1/5 gave rise to increases in phosphorylation of ERK2. Coactivation of mGluR1/5 and DR1/5 with (S)-3,5-dihydroxyphenylglycine and (+)-1-phenyl-2,3,4,5-tetrahydro (1H)-3-benzazepine-7,8-diol hydrochloride enhanced the phosphorylation of ERK2. This interaction between mGluR1/5 and DR1/5 required protein kinase C (PKC), because the PKC inhibitors calphostin C, bisindolylmaleimide I, and Go6976 blocked DR1/5-enhanced phosphorylation of ERK2. Use of the phosphatase inhibitors calyculin and okadaic acid indicated that inhibition of protein phosphatases 1 and 2A dramatically enhanced ERK2 phosphorylation by mGluR1/5. Coactivation of mGluR1/5 and DR1/5 also enhanced cAMP-response element binding protein (CREB) phosphorylation (compared with each receptor agonist alone) but did not enhance CREB-mediated transcriptional activity. Thus, signal transduction pathways activated by DR1/5 and mGluR5 interact to modify downstream events in striatal neurons while retaining numerous regulatory checkpoints. PMID- 15829629 TI - TrkB has a cell-autonomous role in the establishment of hippocampal Schaffer collateral synapses. AB - Neurotrophin signaling has been implicated in the processes of synapse formation and plasticity. To gain additional insight into the mechanism of BDNF and TrkB influence on synapse formation and synaptic plasticity, we generated a conditional knock-out for TrkB using the cre/loxp system. Using three different cre-expressing transgenic mice, three unique spatial and temporal configurations of TrkB deletion were obtained with regard to the hippocampal Schaffer collateral synapse. We compare synapse formation in mutants in which TrkB is ablated either in presynaptic or in both presynaptic and postsynaptic cells at early developmental or postdevelopmental time points. Our results indicate a requirement for TrkB at both the presynaptic and postsynaptic sites during development. In the absence of TrkB, synapse numbers were significantly reduced. In vivo ablation of TrkB after synapse formation did not affect synapse numbers. In primary hippocampal cultures, deletion of TrkB in only the postsynaptic cell, before synapse formation, also resulted in deficits of synapse formation. We conclude that TrkB signaling has a cell-autonomous role required for normal development of both presynaptic and postsynaptic components of the Schaffer collateral synapse. PMID- 15829630 TI - Constitutively active G-protein-gated inwardly rectifying K+ channels in dendrites of hippocampal CA1 pyramidal neurons. AB - A diversity of ion channels contributes to the active properties of neuronal dendrites. From the apical dendrites of hippocampal CA1 pyramidal neurons, we recorded inwardly rectifying K+ channels with a single-channel conductance of 33 pS. The inwardly rectifying K+ channels were constitutively active at the resting membrane potential. The amount of constitutive channel activity was significantly larger in the apical dendrites than in the soma. Activities of these inwardly rectifying K+ channels were inhibited by Ba2+ (200 microM) and tertiapin (10 nM), both of which are believed to block G-protein-coupled inwardly rectifying K+ (GIRK) channels. Intracellularly applied GTPgammaS (20 microM) during dual dendritic recordings significantly increased constitutive channel activity. Baclofen (20 microM), an agonist for the G-protein-coupled GABA(B) receptor, also significantly increased the level of channel activity. Therefore, these channels are GIRK channels, which are constitutively active at rest in the apical dendrites of CA1 pyramidal neurons and can be further activated via G-protein coupled neurotransmitter receptors. PMID- 15829631 TI - Ric-8B, an olfactory putative GTP exchange factor, amplifies signal transduction through the olfactory-specific G-protein Galphaolf. AB - The olfactory system is able to detect a large number of chemical structures with a remarkable sensitivity and specificity. Odorants are first detected by odorant receptors present in the cilia of olfactory neurons. The activated receptors couple to an olfactory-specific G-protein (Golf), which activates adenylyl cyclase III to produce cAMP. Increased cAMP levels activate cyclic nucleotide gated channels, causing cell membrane depolarization. Here we used yeast two hybrid to search for potential regulators for Galphaolf. We found that Ric-8B (for resistant to inhibitors of cholinesterase), a putative GTP exchange factor, is able to interact with Galphaolf. Like Galphaolf, Ric-8B is predominantly expressed in the mature olfactory sensory neurons and also in a few regions in the brain. The highly restricted and colocalized expression patterns of Ric-8B and Galphaolf strongly indicate that Ric-8B is a functional partner for Galphaolf. Finally, we show that Ric-8B is able to potentiate Galphaolf-dependent cAMP accumulation in human embryonic kidney 293 cells and therefore may be an important component for odorant signal transduction. PMID- 15829632 TI - Torsin-mediated protection from cellular stress in the dopaminergic neurons of Caenorhabditis elegans. AB - Parkinson's disease (PD) is linked genetically to proteins that function in the management of cellular stress resulting from protein misfolding and oxidative damage. Overexpression or mutation of alpha-synuclein results in the formation of Lewy bodies and neurodegeneration of dopaminergic (DA) neurons. Human torsinA, mutations in which cause another movement disorder termed early-onset torsion dystonia, is highly expressed in DA neurons and is also a component of Lewy bodies. Previous work has established torsins as having molecular chaperone activity. Thus, we examined the ability of torsinA to manage cellular stress within DA neurons of the nematode Caenorhabditis elegans. Worm DA neurons undergo a reproducible pattern of neurodegeneration after treatment with 6 hydroxydopamine (6-OHDA), a neurotoxin commonly used to model PD. Overexpression of torsins in C. elegans DA neurons results in dramatic suppression of neurodegeneration after 6-OHDA treatment. In contrast, expression of either dystonia-associated mutant torsinA or combined overexpression of wild-type and mutant torsinA yielded greatly diminished neuroprotection against 6-OHDA. We further demonstrated that torsins seem to protect DA neurons from 6-OHDA through downregulating protein levels of the dopamine transporter (DAT-1) in vivo. Additionally, we determined that torsins protect robustly against DA neurodegeneration caused by overexpression of alpha-synuclein. Using mutant nematodes lacking DAT-1 function, we also showed that torsin neuroprotection from alpha-synuclein-induced degeneration occurs in a manner independent of this transporter. Together, these data have mechanistic implications for movement disorders, because our results demonstrate that torsin proteins have the capacity to manage sources of cellular stress within DA neurons. PMID- 15829633 TI - Receptor tyrosine phosphatases guide vertebrate motor axons during development. AB - Receptor-type protein tyrosine phosphatases (RPTPs) are required for appropriate growth of axons during nervous system development in Drosophila. In the vertebrate, type IIa RPTPs [protein tyrosine phosphatase (PTP)-delta, PTP-sigma, and LAR (leukocyte common-antigen-related)] and the type III RPTP, PTP receptor type O (PTPRO), have been implicated in the regulation of axon growth, but their roles in developmental axon guidance are unclear. PTPRO, PTP-delta, and PTP-sigma are each expressed in chick motor neurons during the period of axonogenesis. To examine potential roles of RPTPs in axon growth and guidance in vivo, we used double-stranded RNA (dsRNA) interference combined with in ovo electroporation to knock down RPTP expression levels in the embryonic chick lumbar spinal cord. Although most branches of the developing limb nerves appeared grossly normal, a dorsal nerve identified as the anterior iliotibialis was clearly affected by dsRNA knock-down of RPTPs. In experimental embryos treated with dsRNA targeting PTP-delta, PTP-sigma, or PTPRO, this nerve showed abnormal fasciculation, was reduced in size, or was missing entirely; interference with PTPRO produced the most severe phenotypes. Control embryos electroporated with vehicle, or with dsRNA targeting choline acetyltransferase or axonin-1, did not exhibit this phenotype. Surprisingly, embryos electroporated with dsRNA targeting PTP-delta together with PTPRO, or all three RPTPs combined, had less severe phenotypes than embryos treated with PTPRO alone. This result suggests that competition between type IIa and type III RPTPs can regulate motor axon outgrowth, consistent with findings in Drosophila. Our results indicate that RPTPs, and especially PTPRO, are required for axon growth and guidance in the developing vertebrate limb. PMID- 15829634 TI - cAMP-mediated mechanisms for pain sensitization during opioid withdrawal. AB - Chronic opioid-induced drug dependence and withdrawal syndrome after opioid cessation remain a severe obstacle in clinical treatment of chronic pain and opioid drug addiction. One of the key symptoms during opioid withdrawal is a state of sensitized pain. The most significant molecular adaptation induced by chronic opioids in the brain is upregulation of the cAMP-signaling pathway. Although the cAMP system is known to have multiple effects on central neuron functions, how its upregulation mediates behavioral opioid dependence and withdrawal-induced pain in vivo remains unclear. In this study, we demonstrate that withdrawal from chronic morphine significantly upregulates the mRNA level of adenylyl cyclase (AC) VI and VIII isoforms and immunoreactivity of ACV/VI in the nucleus raphe magnus (NRM), a brainstem site critically involved in opioid modulation of pain. In cellular studies of NRM neurons containing mu-opioid receptors, we show that morphine withdrawal significantly increases glutamate synaptic transmission via a presynaptic mechanism mediated by an upregulated cAMP pathway. Morphine withdrawal also enhances the hyperpolarization-activated current in these neurons by increased intracellular cAMP. Both of the withdrawal induced cAMP actions increase the excitability of these mu-receptor-containing neurons, which are thought to facilitate spinal pain transmission. Furthermore, in morphine-dependent rats in vivo, blocking the cAMP pathway significantly reduces withdrawal-induced pain sensitization. These results illustrate neurobiological mechanisms for the cAMP-mediated withdrawal pain and provide potential therapeutic targets for the treatment of opioid dependence and withdrawal-related problems. PMID- 15829635 TI - Identification of genes involved in synaptogenesis using a fluorescent active zone marker in Caenorhabditis elegans. AB - Active zones are presynaptic regions where synaptic vesicles fuse with plasma membrane to release neurotransmitters. Active zones are highly organized structurally and are functionally conserved among different species. Synapse defective-2 (SYD-2) family proteins regulate active zone morphology in Caenorhabditis elegans and Drosophila. Here, we demonstrate by immunoelectron microscopy that at C. elegans synapses, SYD-2 localizes strictly at active zones and can be used as an active zone marker when fused to green fluorescent protein (GFP). By driving expression of SYD-2::GFP fusion protein in GABAergic neurons, we are able to visualize discrete fluorescent puncta corresponding to active zones in living C. elegans. During development, the number of GABAergic synapses made by specific motoneurons increases only slightly from larvae to adult stages. In contrast, the number of SYD-2::GFP puncta doubles, suggesting that individual synapses accommodate the increasing size of their synaptic targets mainly by incorporating more active zone materials. Furthermore, we used this marker to perform a genetic screen to identify genes involved in the development of active zones. We recovered 16 mutants with altered SYD-2::GFP expression, including alleles of five genes that have been implicated previously in synapse formation or nervous-system development. Mapping of 11 additional mutants suggests that they may represent novel genes involved in active zone formation. PMID- 15829636 TI - Reluctant vesicles contribute to the total readily releasable pool in glutamatergic hippocampal neurons. AB - The size of the readily releasable pool (RRP) of vesicles is critically important for determining the size of postsynaptic currents generated in response to action potentials. However, discrepancies in RRP estimates exist among methods designed to measure RRP size. In glutamatergic hippocampal neurons, we found that hypertonic sucrose application yielded RRP size estimates approximately fivefold larger than values obtained with high-frequency action potential trains commonly assumed to deplete the RRP. This discrepancy was specific for glutamatergic neurons, because no difference was found between sucrose and train estimates of RRP size in GABAergic neurons. A small component of the difference in excitatory neurons was accounted for by postsynaptic receptor saturation. Train estimates of vesicle pool size obtained using more stimuli revealed that action potential elicited EPSCs did not truly reach a steady state during shorter trains, and RRP estimates were closer to sucrose estimates made in the same neurons. This suggested that reluctant vesicles may contribute to the total available pool. Two additional lines of evidence supported this hypothesis. First, RRP estimates from strongly depolarizing hyperkalemic solutions closely matched those obtained with sucrose. Second, when Ca2+ influx was enhanced during trains, train estimates of pool size matched those obtained with sucrose. These data suggest that glutamatergic hippocampal neurons maintain a heterogeneous population of vesicles that can be differentially released with varying Ca2+ influx, thereby increasing the range of potential synaptic responses. PMID- 15829637 TI - Increased capacity and density of choline transporters situated in synaptic membranes of the right medial prefrontal cortex of attentional task-performing rats. AB - Cholinergic neurons innervating the cortex have been conceptualized as a major component of the attention system of the brain. Because of recent evidence indicating plastic mechanisms regulating choline transporter (CHT)-mediated high affinity choline uptake, which is the rate-limiting step of acetylcholine synthesis, the present experiment determined the capacity of cholinergic terminals to transport choline, and the proportion of choline transporters localized in the membrane of synaptic terminals, in several brain regions of rats performing a cognitive vigilance task (CVT) and a simple reaction time task (SRTT) and nonperforming (NP) rats. Compared with evidence from NP rats, increased choline transporter capacity [as indicated by maximum transporter velocity (Vmax)] and an increased density of CHTs situated in synaptic plasma membrane, relative to intracellular locations, were observed in the medial prefrontal cortex of the right but not left hemisphere of CVT-performing animals. Furthermore, right medial prefrontal Vmax values of CVT-performing rats correlated positively and left medial Vmax values correlated negatively with the animals' performance in signal trials. Measures of CHT function in the brains of SRTT-performing animals did not differ significantly from those in NP rats. The present data support the hypothesis that an increased capacity of choline transporters in the right medial prefrontal cortex, primarily attributable to increased trafficking of transporters from intracellular compartments to the terminal membrane, represents a cellular mechanism contributing to the mediation of attentional performance. PMID- 15829638 TI - Feedforward inhibition of projection neurons by fast-spiking GABA interneurons in the rat striatum in vivo. AB - Discharge activities and local field potentials were recorded in the orofacial motor cortex and in the corresponding rostrolateral striatum of urethane anesthetized rats. Striatal projection neurons were identified by antidromic activation and fast-spiking GABAergic interneurons (FSIs) by their unique characteristics: briefer spike and burst responses. Juxtacellular injection of neurobiotin combined with parvalbumin immunohistochemistry validated this identification. Spontaneous activities and spike responses to cortical stimulation were recorded during both states of cortical activity: slow waves and desynchronization. Both FSI and projection neurons spontaneously discharged synchronously with slow waves at the maximum of cortical activity, but, on average, FSIs were much more active. Cortical desynchronization enhanced FSI activity and facilitated their spike responses to cortical stimulation, whereas opposite effects were observed regarding projection neurons. Experimental conditions favoring FSI discharge were always associated with a decrease in the firing activity of projection neurons. Spike responses to cortical stimulation occurred earlier (latency difference, 4.6 ms) and with a lower stimulation current for FSIs than for projection neurons. Moreover, blocking GABA(A) receptors by local picrotoxin injection enhanced the spike response of projection neurons, and this increase was larger in experimental conditions favoring FSI responses. Therefore, on average, FSIs exert in vivo a powerful feedforward inhibition on projection neurons. However, a few projection neurons were actually more sensitive to cortical stimulation than FSIs. Moreover, picrotoxin, which revealed FSI inhibition, preferentially affected projection neurons exhibiting the weakest sensitivity to cortical stimulation. Thus, feedforward inhibition by FSIs filters cortical information effectively transmitted by striatal projection neurons. PMID- 15829639 TI - Postpubertal sex differentiation of forebrain structures and functions depend on transforming growth factor-alpha. AB - Sex- and age-associated deficits in brain structure and behavior are reported in a number of neuropsychiatric disorders. Although genetic and environmental factors are thought to contribute to the pathogenesis, there are only few examples in clinical or experimental systems that have identified specific causes. Here, we report that transforming growth factor-alpha (TGFalpha) may regulate sex- and age-dependent development of forebrain structures and associated neural functions after puberty. Waved-1 (Wa-1) mice inherit an autosomal recessive, spontaneous mutation that results in a postnatal reduction in TGFalpha gene expression. The assessment of forebrain structures using a three dimensional magnetic resonance microscopy indicated ventricular enlargement and striatal reduction in both male and female Wa-1 adult mice, with Wa-1 males exhibiting a more severe phenotype. In contrast, the hippocampal volume was reduced only in adult Wa-1 males. Similarly, behavioral analyses showed impaired auditory and contextual fear learning in adult Wa-1 males only, whereas abnormal stress response was expressed by both male and female adult Wa-1 mice. Interestingly, all behavioral deficits were absent before full sexual maturation, despite some slight forebrain structural abnormalities. These results suggest that TGFalpha may regulate postpubertal, sex differentiation in ventricular and periventricular anatomy and associated behavior, affecting predominantly males. In particular, the adult male-specific reduction in hippocampal volume may reflect an age- and sex-specific regulation of stress homeostasis and fear learning. Furthermore, a lack of a behavioral phenotype, despite anatomical alterations in peripubertal Wa-1 mice, suggests that analysis of certain neuroanatomical features at puberty may predict neurobehavioral deficits in adulthood. PMID- 15829640 TI - Role of amygdalo-nigral circuitry in conditioning of a visual stimulus paired with food. AB - The amygdala central nucleus (CeA) plays an important part in associative learning. Although most research has focused on functions of its descending projections to brainstem areas involved in autonomic and somatomotor responses, the ascending projections of CeA also play critical roles in learning. For example, a CeA-nigrostriatal pathway is important for acquiring orienting responses (ORs) to conditioned stimuli (CSs) that signal food delivery. In this study, the function of this CeA-nigrostriatal pathway in appetitive conditioning of rats was considered in more detail. In experiment 1, we combined anatomical tracing and methods for detecting neuronal activation to examine whether CeA neurons that project to the substantia nigra pars compacta (SNc) are activated by a visual CS for food. After injection of the retrograde tracer Fluoro-Gold (FG) into SNc, the rats received pairings of a visual CS with food. After a test with the CS alone, the brains were prepared to assess FG labeling and CS-induced Fos expression in CeA with immunohistochemical procedures. Colocalization of Fos and FG in CeA neurons was visualized with confocal-fluorescence microscopy. The CS induced Fos expression in CeA, and a majority of these Fos-positive neurons were also FG positive, indicating activation of the CeA-SNc pathway by the CS. In experiment 2, lesions that disconnected CeA and SNc prevented the acquisition of conditioned ORs but did not affect the acquisition of conditioned food-related responses or the display of unconditioned ORs. These experiments demonstrate a role for amygdalo-nigral circuitry in learned modulation of attention to signals for biologically significant events. PMID- 15829641 TI - Downregulation of activating transcription factor 5 is required for differentiation of neural progenitor cells into astrocytes. AB - The mechanisms that regulate neural progenitor cell differentiation are primarily unknown. The transcription factor activating transcription factor 5 (ATF5) is expressed in neural progenitors of developing brain but is absent from mature astrocytes and neurons. Here, we demonstrate that ATF5 regulates the conversion of ventricular zone (VZ) and subventricular zone (SVZ) neural progenitors into astrocytes. Constitutive ATF5 expression maintains neural progenitor cell proliferation and blocks their in vitro and in vivo differentiation into astrocytes. Conversely, loss of ATF5 function promotes cell-cycle exit and allows astrocytic differentiation in vitro and in vivo. CNTF, a promoter of astrocytic differentiation, downregulates endogenous ATF5, whereas constitutively expressed ATF5 suppresses CNTF-promoted astrocyte genesis. Unexpectedly, constitutive ATF5 expression in neonatal SVZ cells both in vitro and in vivo causes them to acquire properties and anatomic distributions of VZ cells. These findings identify ATF5 as a key regulator of astrocyte formation and potentially of the VZ to SVZ transition. PMID- 15829642 TI - Modular organization of excitatory circuits between neurons of the spinal superficial dorsal horn (laminae I and II). AB - Neural circuitry of the spinal superficial dorsal horn (SDH) (laminae I and II) and its relationship to pain and other somatosensory phenomena remain poorly understood. To gain information on this issue, synaptic connections between identified SDH neurons were studied in rat spinal cord slices by simultaneous whole-cell recordings from pairs of cells. Both excitatory and inhibitory connections were noted. This report focuses on the observed excitatory linkages. Synaptic excitatory connections between SDH neurons proved highly selective and consistently were unidirectional. Two patterns repeatedly appeared (for neuron classification, see Materials and Methods) (Grudt and Perl, 2002). Lamina II central neurons, with dorsal root (DR) C-fiber input, monosynaptically excited lamina II vertical neurons with DR Adelta input. Lamina II outer vertical neurons with DR Adelta input monosynaptically excited lamina I neurons. Some of the postsynaptic lamina I cells were shown to project rostrally. In contrast to the usual case for connected neurons, in unconnected pairs, primary afferent input to the same type of neuron proved closely similar. Together, these observations indicate that the neural circuitry in the SDH, including its substantia gelatinosa (lamina II), has an explicit organization in which particular combinations of neurons comprise modules arranged to modify and transmit sensory information arriving from Adelta and C primary afferent fibers. PMID- 15829644 TI - Timing of conditioned eyeblink responses is impaired in cerebellar patients. AB - In the present study, timing of conditioned eyeblink responses (CRs) was investigated in cerebellar patients and age-matched controls using a standard delay paradigm. Findings were compared with previously published data of CR incidences in the same patient population (Gerwig et al., 2003; Timmann et al., 2005). Sixteen patients with pure cortical cerebellar degeneration (spinocerebellar ataxia type 6 and idiopathic cerebellar ataxia), 14 patients with lesions within the territory of the superior cerebellar artery, and 13 patients with infarctions within the territory of the posterior inferior cerebellar artery were included. The affected cerebellar lobules and possible involvement of cerebellar nuclei were determined by three-dimensional magnetic resonance imaging (MRI) in patients with focal lesions (n = 27). Based on a voxel by-voxel analysis, MRI lesion data were related to eyeblink conditioning data. CR incidence was significantly reduced, and CRs occurred significantly earlier in patients with cortical cerebellar degeneration and lesions of the superior cerebellum compared with controls. Incidence and timing of CRs was not impaired in patients with lesions restricted to the posterior and inferior cerebellum. Voxel-based MRI analysis revealed that cortical areas within the anterior lobe (Larsell lobule HV) were most significantly related to timing deficits, whereas reduced CR incidences were related to more caudal parts (lobule HVI) of the superior cerebellar cortex. The present data suggest that different parts of the superior cerebellar cortex may be involved in the formation of the stimulus association and appropriate timing of conditioned eyeblink responses in humans. Extracerebellar premotoneuronal disinhibition, however, is another possible explanation for changes in CR timing. PMID- 15829643 TI - Hearing loss raises excitability in the auditory cortex. AB - Developmental hearing impairments compromise sound discrimination, speech acquisition, and cognitive function; however, the adjustments of functional properties in the primary auditory cortex (A1) remain unknown. We induced sensorineural hearing loss (SNHL) in developing gerbils and then reared the animals for several days. The intrinsic membrane and synaptic properties of layer 2/3 pyramidal neurons were subsequently examined in a thalamocortical brain slice preparation with whole-cell recordings and electron microscopic immunocytochemistry. SNHL neurons displayed a depolarized resting membrane potential, an increased input resistance, and a higher incidence of sustained firing. They also exhibited significantly larger thalamocortically and intracortically evoked excitatory synaptic responses, including a greater susceptibility to the NMDA receptor antagonist AP-5 and the NR2B subunit antagonist ifenprodil. This correlated with an increase in NR2B labeling of asymmetric synapses, as visualized ultrastructurally. Furthermore, decreased frequency and increased amplitude of miniature EPSCs (mEPSCs) in SNHL neurons suggest that a decline in presynaptic release properties is compensated by an increased excitatory response. To verify that the increased thalamocortical excitation was elicited by putative monosynaptic connections, minimum amplitude ventral medial geniculate nucleus-evoked EPSCs were recorded. These minimum evoked responses were of larger amplitude, and the NMDAergic currents were also larger and longer in SNHL neurons. These findings were supported by significantly longer AP-5-sensitive durations and larger amplitudes of mEPSCs. Last, the amplitudes of intracortically evoked monosynaptic and polysynaptic GABAergic inhibitory synaptic responses were significantly smaller in SNHL neurons. These alterations in cellular properties after deafness reflect an attempt by A1 to sustain an operative level of cortical excitability that may involve homeostatic mechanisms. PMID- 15829645 TI - Activation of orbital and medial prefrontal cortex by methylphenidate in cocaine addicted subjects but not in controls: relevance to addiction. AB - Drugs of abuse are rewarding to addicted and nonaddicted subjects, but they trigger craving and compulsive intake only in addicted subjects. Here, we used positron emission tomography (PET) and [18F] deoxyglucose to compare the brain metabolic responses (marker of brain function) of cocaine-addicted subjects (n = 21) and controls (n = 15) to identify brain regions that are uniquely activated in addicted subjects by intravenous methylphenidate (a drug that cocaine-addicted subjects report to be similar to cocaine). In parallel, we also measured the changes in dopamine (DA) induced by intravenous methylphenidate (using PET and [11C] raclopride) in the striatum and in the thalamus. Metabolic responses between groups differed significantly only in the right medial orbital prefrontal cortex [Brodmann's area (BA) 25 and medial BA 11], where methylphenidate increased metabolism in addicted subjects but decreased metabolism in controls. These changes were associated in all subjects with increased "desire for methylphenidate" and in the addicted subjects with "cocaine craving." In addicted subjects, increases in BA 25 were also associated with mood elevation. Methylphenidate-induced increases in metabolism in the medial orbital prefrontal cortex were associated with its increase of DA in the thalamus but not in the striatum. These findings provide evidence that enhanced sensitivity of BA 25 (region involved with emotional reactivity) and BA 11 (region involved with salience attribution and motivation) in cocaine-addicted subjects may underlie the strong emotional response to the drug and the intense desire to procure it that results in craving and compulsive drug intake. It also suggests that the mesothalamic DA pathway may contribute to these processes. PMID- 15829646 TI - A strict correlation between dendritic and somatic plateau depolarizations in the rat prefrontal cortex pyramidal neurons. AB - One of the fundamental problems in neurobiology is to understand the cellular mechanism for sustained neuronal activity (neuronal UP states). Prefrontal pyramidal neurons readily switch to a long-lasting depolarized state after suprathreshold stimulation of basal dendrites. Analysis of the dendritic input output function revealed that basal dendrites operate in a somewhat binary regimen (DOWN or UP) in regard to the amplitude of the glutamate-evoked electrical signal. Although the amplitude of the dendritic potential quickly becomes saturated (dendritic UP state), basal dendrites preserve their ability to code additional increase in glutamatergic input. Namely, after the saturation of the plateau amplitude, an additional increase in excitatory input is interpreted as an increase in plateau duration. Experiments performed in tetrodotoxin indicate that the maintenance of a stable depolarized state does not require inhibitory inputs to "balance" the excitation. In the absence of action potential dependent (network-driven) GABAergic transmission, pyramidal neurons respond to brief (5 ms) glutamate pulses with stable long-lasting (approximately 500 ms) depolarizations. Voltage-sensitive dye recordings revealed that this somatic plateau depolarization is precisely time-locked with the regenerative dendritic plateau potential. The somatic plateau rises a few milliseconds after the onset of the dendritic transient and collapses with the breakdown of the dendritic plateau depolarization. In our in vitro model, the stable long-lasting somatic depolarization (UP state like) is a direct consequence of the local processing of a strong excitatory glutamatergic input arriving on the basal dendrite. The slow component of the somatic depolarization accurately mirrors the glutamate-evoked dendritic plateau potential (dendritic UP state). PMID- 15829647 TI - Optical current source density analysis in hippocampal organotypic culture shows that spreading depression occurs with uniquely reversing currents. AB - Spreading depression (SD) involves current flow through principal neurons, but the pattern of current flow over the expanse of susceptible tissues or individual principal neurons remains undefined. Accordingly, tissue and single cell maps made from digital imaging of voltage-sensitive dye changes in hippocampal organotypic cultures undergoing SD were processed via optical current source density analysis to reveal the currents associated with pyramidal neurons. Two distinctive current flow patterns were seen. The first was a trilaminar pattern (420 microm2) that developed with the onset of SD in CA3 pyramidal neurons, in which SD most often began. This initial pattern comprised a somatic current sink with current sources to either side in the dendrites that lasted for seconds extending into the first aspect of the classical "inverted saddle" interstitial direct current waveform of SD. Next, the somatic sink backpropagated at a speed of millimeters per minute into the proximal dendrites, resulting in a reversal of the initial current flow pattern to its second orientation, namely dendritic sinks associated with a somatic source. The latter persisted for the remainder of SD in CA3 and was the only pattern seen in CA1, in which SD was rarely initiated. This backpropagating SD current flow resembles that of activity-dependent synaptic activation. Retrograde and associative signaling via principal neuron current flow is a key means to affect tissue function, including synaptic activation and, by extension, perhaps SD. Such current-related postsynaptic signaling might not only help explain SD but also neuroprotection and migraine, two phenomena increasingly recognized as being related to SD. PMID- 15829648 TI - Phase synchrony among neuronal oscillations in the human cortex. AB - Synchronization of neuronal activity, often associated with network oscillations, is thought to provide a means for integrating anatomically distributed processing in the brain. Neuronal processing, however, involves simultaneous oscillations in various frequency bands. The mechanisms involved in the integration of such spectrally distributed processing have remained enigmatic. We demonstrate, using magnetoencephalography, that robust cross-frequency phase synchrony is present in the human cortex among oscillations with frequencies from 3 to 80 Hz. Continuous mental arithmetic tasks demanding the retention and summation of items in the working memory enhanced the cross-frequency phase synchrony among alpha (approximately 10 Hz), beta (approximately 20 Hz), and gamma (approximately 30-40 Hz) oscillations. These tasks also enhanced the "classical" within-frequency synchrony in these frequency bands, but the spatial patterns of alpha, beta, and gamma synchronies were distinct and, furthermore, separate from the patterns of cross-frequency phase synchrony. Interestingly, an increase in task load resulted in an enhancement of phase synchrony that was most prominent between gamma- and alpha-band oscillations. These data indicate that cross-frequency phase synchrony is a salient characteristic of ongoing activity in the human cortex and that it is modulated by cognitive task demands. The enhancement of cross-frequency phase synchrony among functionally and spatially distinct networks during mental arithmetic tasks posits it as a candidate mechanism for the integration of spectrally distributed processing. PMID- 15829649 TI - Differential responses of bladder lumbosacral and thoracolumbar dorsal root ganglion neurons to purinergic agonists, protons, and capsaicin. AB - The present study explored differences in sensitivity to purinergic agonists, protons, and capsaicin in lumbosacral (LS) and thoracolumbar (TL) sensory neurons that innervate the rat urinary bladder. The majority of LS neurons (93%) were sensitive to alpha,beta-methyleneATP (alpha,beta-metATP) compared with 50% of TL neurons. Based on inactivation kinetics, a slowly desensitizing current evoked by alpha,beta-metATP predominated in LS neurons (86%) compared with mixed components that characterized TL neuron responses (58%). The density of the slowly desensitizing current was greater in LS than in TL neurons (LS, 34.4 +/- 5.3 pA/pF; TL, 2.5 +/- 0.8 pA/pF). Almost all neurons in both ganglia responded to protons and to capsaicin (LS, 100%; TL, 98%). Proton-activated currents in bladder sensory neurons exhibited distinct inactivation kinetics as fast, intermediate, slowly desensitizing, and sustained components. More than one component was expressed in every cell. Although there was no difference in the percentage of neurons expressing more than one component, the density of the sustained current was significantly greater in LS than in TL neurons (LS, 86.1 +/ 16 pA/pF; TL, 30.3 +/- 7 pA/pF). Similarly, the capsaicin-evoked current was greater in LS than in TL neurons (LS, 129.6 +/- 17 pA/pF; TL, 86.9 +/- 11 pA/pF). Finally, a greater percentage of TL neurons bound isolectin B4 than LS neurons (LS, 61%; TL, 85%). The greater degree of alpha,beta-metATP, proton, and capsaicin responsiveness, in addition to differences in current type and current densities, in LS and TL neurons suggests that bladder pelvic and hypogastric/lumbar splanchnic afferents are functionally distinct and likely mediate different sensations arising from the urinary bladder. PMID- 15829650 TI - Sharing receptive fields with your neighbors: tuning the vertical system cells to wide field motion. AB - In the blowfly, the direction-selective response of the 60 lobula-plate tangential cells has been ascribed to the integration of local motion information across their extensive dendritic trees. Because the lobula plate is organized retinotopically, the receptive fields of the tangential cells ought to be determined by their dendritic architecture. However, this appears not always to be the case. One compelling example is the exceptionally wide receptive fields of the vertical system (VS) tangential cells. Using dual-intracellular recordings, Haag and Borst (2004) found VS cells to be mutually coupled in such a way that each VS cell is connected exclusively to its immediate neighbors. This coupling may form the basis of the broad receptive fields of VS cells. Here, we tested this hypothesis directly by photoablating individual VS cells. The receptive field width of VS cells indeed narrowed after the ablation of single VS cells, specifically depending on whether the receptive field of the ablated cell was more frontal or more posterior to the recorded cell. In particular, the responses changed as if the neuron lost access to visual information from the ablated neuron and those VS cells more distal than it from the recorded neuron. These experiments provide strong evidence that the lateral connections among VS cells are a crucial component in the mechanism underlying their complex receptive fields, augmenting the direct columnar input to their dendrites. PMID- 15829651 TI - Letter to the editor: concerning an article comparing the efficacy of local and systemic treatment of clinical mastitis. PMID- 15829652 TI - Beta-lactoglobulin is a thermal marker in processed milk as studied by electrophoresis and circular dichroic spectra. AB - As much of the sterilization process involves heat treatment during the preparation of milk on an industrial scale, the unpredictable measures of the process are an essential issue in determining the quality of the milk. The purpose of the present study was to investigate the major protein change(s) of whey proteins in processed milk and extend the knowledge for future reference in the dairy industry. Using a native polyacrylamide gel electrophoresis, we showed almost a 90% loss and denaturation of beta-lactoglobulin (LG), but not alpha lactalbumin (LA), in some brands of the processed and dry milks. Immunochemical analysis using Western blotting revealed that part of the loss was attributed to the formation of large multiple forms of LG in the processed product. Such denaturation was presumably associated with the heating procedure used in the process. Essentially, LG was the only major fraction converted to aggregates in milk heated at 95 degrees C for 30 min on 2-dimensional PAGE. The detailed thermal denaturation of purified LG and LA at various temperatures (50 to 95 degrees C) and time (5 to 960 s) were investigated using a circular dichroic analysis. The maximal changes of ellipticity at 205 nm (converting beta-structure to disordered structure) were correlated to heating temperature and time. There were no significant conformational changes of LG at temperatures below 70 degrees C for as long as 480 s. Pronounced and rapid changes occurred between 80 to 95 degrees C in a time-dependent manner. Fifty percent of the maximal changes could be reached within 15 s. In conclusion, the unique chemical and immunochemical loss and conformational changes made LG a superior marker for evaluating the thermal processing of milk. The detailed thermal denaturation curves of LG constructed with its time and temperature in this study provide a valuable reference for the dairy industry. We postulate that heat treatment over 80 degrees C in 15 s may induce a significant denaturation of milk LG. PMID- 15829653 TI - Short communication: rearrangement of rumenic Acid in ruminant fats: a marker of thermal treatment. AB - Rumenic (cis-9,trans-11 18:2) acid is the main conjugated linoleic acid (CLA) isomer in milk and other ruminant fats. Anhydrous regular and high-CLA butterfats were heated at 200 degrees C for 2, 4, and 6 h under atmospheric conditions. [1,5] Sigmatropic isomerization of rumenic acid occurred, resulting in the formation of trans-8,cis-10 18:2 acid, as determined by mass spectrometry of its 4,4-dimethyloxazoline derivative. Rate of isomerization was monitored by gas liquid chromatography, using a 120-m capillary column coated with 70% equivalent cyanoalkylpolysiloxane polymer, and reaction was of first order. Furthermore, [1,5] sigmatropic rearrangement product analysis can be used as an indicator of heat treatment of natural fats and oils containing CLA. PMID- 15829654 TI - Rate of maillard browning in sweet whey powder. AB - The objective of this study was to evaluate the rate of Maillard browning in 3 commercial sweet whey powders (WC1, WC2, and MW1), under accelerated shelf-life testing (ASLT) and under normal storage conditions (21 degrees C and 35% RH). Rate of brown pigment formation (k) obtained from short-term ASLT of whey powder was compared with actual findings obtained from the long-term shelf-life testing under normal conditions. Deterioration by Maillard browning, measured by spectrophotometer, was compared with changes in color (Hunter Laboratory), free moisture, titratable acidity, and sensory attributes. Results suggest that estimated k (from ASLT) was comparable with the observed rate (obtained at ambient temperature) for 2 producers (WC1, MW1). The actual k values observed for samples WC1, WC2, and MW1, stored under normal conditions, were 0.0031, 0.0080, and 0.0148 color units/g of solid per mo, respectively. The estimated values of k for samples WC1, WC2, and MW1 were 1.12, 4.90, and 1.35 times more than the observed values, respectively. The Q10 values (increase in reaction rate for a 10 degrees C temperature increase) ranged from 1.77 to 4.14, and the activation energies ranged from 15.9 to 28.4 kcal/mol. Hunter Laboratory values L* and a* appeared most sensitive to changes during storage. Free moisture content, and acidity increased significantly with storage. However, no significant changes were detected by the sensory panel in the attributes considered. PMID- 15829655 TI - The kinetics of heat-induced structural changes of beta-lactoglobulin. AB - Heat-induced structural changes of beta-lactoglobulin were studied at temperatures ranging from 67.5 to 82.5 degrees C, and at pH 7.5. These changes were monitored by measurement of surface hydrophobicity, thiol availability, and protein solubility. Kinetic studies were conducted to quantitatively describe the contribution of hydrophobic and SH/SS interchange reactions to the thermal structural changes of beta-lactoglobulin. Results indicate that beta lactoglobulin is sensitive to heat-induced interchange reactions with consequences for protein solubility. The extent of changes measured by the increase in surface hydrophobicity and the decrease in slow-reacting SH groups content could be described by a first-order fractional conversion model and were characterized by activation energy values of 233.9 +/- 8.6 and 148.2 +/- 6.7 kJ/mol, respectively. The break in the Arrhenius plot suggested in literature for beta-lactoglobulin denaturation was confirmed in this study only for the kinetics of exposed SH groups. PMID- 15829656 TI - The balance between caseins and whey proteins in cow's milk determines its allergenicity. AB - Cow's milk allergy is quite common in the first years of human life. Protein composition plays an important role in this pathology, particularly the casein/whey protein ratio. It is known that milks from different species have different sensitization capacities although their protein sources are quite similar. Thus, the objective of this work was to compare the allergenicity of native cow's milk and milk with a modified ratio of casein and whey proteins in a murine model of atopy. Twenty-four Balb/c mice were orally sensitized to native cow's milk or modified cow's milk with a casein/whey protein ratio of 40:60. During the sensitization period, the number of mice suffering from diarrhea was significantly higher in the native cow's milk-sensitized group than in the modified milk-sensitized group. Once mice were killed, plasma histamine levels were shown to be significantly higher in native cow's milk-sensitized mice. In addition, cow's milk proteins induced a higher lymphocyte sensitization in the native milk-sensitized mice, with a significant increase in the specific proliferation ratio of these cells. These results suggest that the balance between caseins and whey proteins plays an important role in the sensitization capacity of cow's milk, and its modification might be a way to reduce the allergenicity of cow's milk. PMID- 15829657 TI - Storage stability of lutein during ripening of cheddar cheese. AB - Lutein (3,3'-dihydroxy-alpha-carotene) has been identified as a dietary factor that can delay the onset of age-related macular degeneration (AMD). However, available food sources of lutein contain only modest amounts of the carotenoid. Food fortification with lutein extract has been identified as a low-budget approach to prevent the onset or progression of AMD. The objectives of this study were to 1) incorporate various amounts of lutein into Cheddar cheese; 2) examine the color, pH, microbiological, and sensory characteristics of the Cheddar cheese during storage; and 3) analyze the stability of lutein during the cheese maturation process. Lutein extracted from corn was added to Cheddar cheese in quantities of 1, 3, and 6 mg per serving size. Measurements of the lutein stability were carried out by HPLC using a YMC C30 carotenoid column. Microbiological analyses of cheese samples included aerobic plate count, coliform, and yeast/mold counts. The color attributes a* and b* were significantly different between the treatment and control groups; however, no significant difference was observed in L* value and pH. Significant differences among 1, 3, and 6 mg lutein-enriched cheeses were observed in the aerobic plate count and yeast/mold compared with the control. Cheese samples contained no detectable levels of coliforms (< 10 cfu/g). The HPLC data showed quantitative recovery of lutein during the storage period, and no lutein degradation products were identified. These results indicate that lutein, a functional additive with purported ability to prevent or reduce the onset of AMD, can be incorporated into cheese adding value to this product. PMID- 15829658 TI - An iterative sensory procedure to select odor-active associations in complex consortia of microorganisms: application to the construction of a cheese model. AB - The aim of this study was to develop and validate an iterative procedure based on odor assessment to select odor-active associations of microorganisms from a starting association of 82 strains (G1), which were chosen to be representative of Livarot cheese biodiversity. A 3-step dichotomous procedure was applied to reduce the starting association G1. At each step, 3 methods were used to evaluate the odor proximity between mother (n strains) and daughter (n/2 strains) associations: a direct assessment of odor dissimilarity using an original bidimensional scale system and 2 indirect methods based on comparisons of odor profile or hedonic scores. Odor dissimilarity ratings and odor profile gave reliable and sometimes complementary criteria to select G3 and G4 at the first iteration, G31 and G42 at the second iteration, and G312 and G421 at the final iteration. Principal component analysis of odor profile data permitted the interpretation at least in part, of the 2D multidimensional scaling representation of the similarity data. The second part of the study was dedicated to 1) validating the choice of the dichotomous procedure made at each iteration, and 2) evaluating together the magnitude of odor differences that may exist between G1 and its subsequent simplified associations. The strategy consisted of assessing odor similarity between the 13 cheese models by comparing the contents of their odor-active compounds. By using a purge-and-trap gas chromatography olfactory/mass spectrometry device, 50 potent odorants were identified in models G312, G421, and in a typical Protected Denomination of Origin Livarot cheese. Their contributions to the odor profile of both selected model cheeses are discussed. These compounds were quantified by purge and trap-gas chromatography mass spectrometry in the 13 products and the normalized data matrix was transformed to a between-product distance matrix. This instrumental assessment of odor similarities allowed validation of the choice of G312 as the best 10-strain ecosystem. PMID- 15829659 TI - Comparison of calcium salts and formaldehyde-protected conjugated linoleic Acid in inducing milk fat depression. AB - Abomasal infusion studies have shown that trans-10, cis-12 conjugated linoleic acid (CLA) decreases milk fat synthesis. However, supplements of CLA must avoid rumen biohydrogenation for this technology to be applied to ruminants. Rumen protection methods would reduce CLA metabolism in the rumen and increase its supply to the small intestine. Our objective was to compare the efficacy of 2 forms of rumen-protected CLA at inducing milk fat depression. Three mid to late lactation Holstein cows each fitted with a rumen fistula were used in a 3 x 3 Latin square design. Treatments were: 1) control, 2) calcium salts of CLA (Ca CLA), and 3) formaldehyde-protected CLA (FP-CLA). Supplements were designed to provide 10 g/d of trans-10, cis-12 CLA and were administered intraruminally once per day to ensure exact delivery of amount. Both CLA treatments substantially reduced milk fat yield and content compared with control, with the reductions in milk fat yield averaging 34% for the Ca-CLA treatment and 44% for the FP-CLA treatment. In contrast, milk yield, milk protein yield, and dry matter intake were unaltered by CLA treatment. Efficiency of transfer of trans-10, cis-12 CLA from the supplement into milk fat was 3.2 and 7.0% for Ca-CLA and FP-CLA, respectively. These values are much lower than transfer efficiencies reported for abomasally infused CLA, suggesting that much of the trans-10, cis-12 CLA present in the 2 formulations was biohydrogenated in the rumen. Overall, the extent of the reduction in milk fat yield indicates that both protection formulations are acceptable methods for the formulation of CLA supplements to induce milk fat depression in lactating dairy cows. PMID- 15829660 TI - Subclinical mastitis and antimicrobial susceptibility of Staphylococcus caprae and Staphylococcus epidermidis isolated from two Italian goat herds. AB - A total of 156 goats from 2 commercial dairy goat farms were monitored for intramammary infections during an entire lactation. Most of the infections (80.7%) observed were due to coagulase-negative staphylococci (CNS) species. In herd 1, nearly all of the infections (96%) were due to CNS species, with Staphylococcus caprae (SCAP) being the most common specific pathogen observed, accounting for about 43% of the infections. In herd 2, the proportion of the infections due to CNS was 67% and Staphylococcus epidermidis (SEPI) was the most common pathogen (48% of infections) and SCAP was not present. Linear somatic cell scores (SCS) were greater in milk from infected udder halves, with an average difference of 0.78 SCS. The SCS for infected udder halves was greater than noninfected for all CNS species, although differences among species were observed. The ranking across CNS species was SCAP > other CNS > SEPI > no infection. However, infections by SEPI tended to be more persistent. Increased SCS was associated with a statistically significant decrease in milk yield, but no effect was observed for intramammary infections (IMI). Ninety-seven isolates of CNS (53 SCAP and 44 SEPI) were investigated for in vitro susceptibility to several antimicrobial agents. Benzylpenicillin was the most effective antimicrobial agent against SCAP and SEPI. A concentration of 0.05 microg/mL was sufficient to inhibit growth of 90% of SCAP colonies, and 0.10 microg/mL yielded a similar effectiveness for SEPI. Amoxicillin and the combination of amoxicillin and clavulanic acid were only slightly less effective. Tetracycline (62.5 microg/mL) and tilmicosin (500 microg/mL) were the least effective treatments for SEPI and SCAP, respectively. PMID- 15829661 TI - Technical note: molecular typing of Corynebacterium bovis isolates by pulsed field gel electrophoresis. AB - A typing method for Corynebacterium bovis based on pulsed-field gel electrophoresis (PFGE) is described and applied to type 162 strains isolated from mastitic milk from 57 Friesian dairy herds in northern Spain. Isolates were identified to the species level using the API-Coryne system and a polymerase chain reaction-restriction fragment length polymorphism of the 16S rDNA amplicon. Cultures in logarithmic phase were harvested, mixed with agarose, and treated with lisozyme and proteinase K to release intact genomic DNA that was digested with XbaI and submitted to PFGE. Seven different pulsotypes were identified; none of them were similar to that of the reference strain (ATCC 7715). The most frequent pulsotype was pattern I (51.9%) followed by pattern II (29.6%). The majority of the herds showed infection with one single pulsotype, but 12 herds yielded 2 pulsotypes, and one herd had 3 pulsotypes. No relationship was found between the geographical location of the herd and the pulsotype. This is the first description of a typing method for C. bovis; the technique might prove useful for epidemiological studies on this species. PMID- 15829662 TI - Short communication: hydroperoxides in circulating lipids from dairy cows: implications for bioactivity of endogenous-oxidized lipids. AB - This study was conducted to investigate the potential for increased oxidative stress of high- vs. average-producing dairy cows. Two experiments were performed using 11 and 13 Holstein cows (53 +/- 2 d postpartum). Lipohydroperoxides (LHP) were determined in serum lipids (experiment 1) and low-density lipoprotein (experiment 2) via oxidation of ferrous to ferric ions through LHP using thiocyanate as chromogen. In experiment 1, differing milk yield and milk energy output corresponded to different concentrations of LHP. In experiment 2, analysis of regression resulted in a significant relationship between milk yield and LHP. Phospholipids isolated from lipids with 6.5 microM of LHP evoked in monocytic cells a transient increase in superoxide formation, indicating inflammatory potential. The results show that high milk productivity can associate with oxidative stress indicated by oxidative modifications of circulating lipids and their changed bioactivity. PMID- 15829663 TI - Automatic milking and grazing--effects of location of drinking water on water intake, milk yield, and cow behavior. AB - In an automatic milking system with cows grazing on a mixed grass sward, experiments were performed in 2001 and 2003, lasting 10 and 7 wk, respectively. Two location strategies for offering drinking water were compared: available in the barn and in the field (group B+F) or only in the barn (group B). During 2001, cows grazed alternately at 2 pastures at different distances from the barn, 50 m (near pasture) or 330 m (distant pasture), whereas the distant pasture was mainly used in 2003. No significant differences in milk yield, milking frequency, or milk composition were found between the 2 treatments. Average milk yield in the 2 experiments was 26.8 and 27.6 kg of milk in 2001 and 2003, respectively, and average milking frequency was 2.4 milkings/ d. Significant differences in animal behavior were observed only during the period when animals grazed on the distant pasture in 2001, with animals in group B+F spending 40% of their time on pasture and 21% of their time grazing, whereas corresponding values for group B were 34 and 17%, respectively. In 2003, average drinking water intakes per cow were 53 L/d on treatment B and 51 L/d on treatment B+F, and were not significantly different. Total daily water intake including water in the pasture was approximately 90 L/cow. In conclusion, no significant differences in milk yield, milking frequency, or water intake were found between cows offered drinking water both in the barn and in the field compared with drinking water only in the barn at pasture distances up to 330 m. PMID- 15829664 TI - Growth hormone response to growth hormone releasing hormone in calves that differ in genetic merit for milk yield. AB - Holstein heifer, steer, and bull calves from control (CL) and select (SL) lines of cows that differed by more than 4000 kg of milk during a 305-d lactation (SL > CL) were used to determine growth hormone (GH) response to 5 doses of GH releasing hormone (GHRH) and how this response was affected by gender, period (age), and genetic merit for milk yield. Doses (0, 2.5, 5, 10, and 20 microg/100 kg of BW) of a GHRH analog were assigned randomly to each heifer (4 CL, 4 SL), steer (4 CL, 4 SL), and bull (3 CL, 3 SL) calf and administered on consecutive days at approximately 3, 6, and 10 mo of age (periods; P1, P2, and P3). Jugular blood samples (n = 15) collected between -30 and 240 min relative to GHRH administration were used to quantify area under the GH response curve (AUC) after subtracting mean prechallenge GH concentrations. Estimates of maximum response (Rmax) and sensitivity (ED50) to GHRH were obtained from the hyperbolic dose response curves (AUC vs. dose). Data were analyzed for effects of dose, line, period, gender, and their interactions with period as the repeated effect. Prechallenge GH concentrations were not affected by genetic line, gender, or period. The AUC was not affected by line, but decreased with period and increased with GHRH dose. The Rmax did not differ between lines or among genders, but decreased with period. The ED50 did not differ between lines or among periods, but heifers were more sensitive to GHRH than steers or bulls. Although GH response to GHRH has been identified as a potential indicator of genetic merit, it did not differ between these substantially different genetic lines. PMID- 15829665 TI - Effect of supplemental conjugated linoleic acids on heat-stressed brown swiss and holstein cows. AB - Heat-stressed dairy cattle are bioenergetically similar to early-lactation cows in that dietary energy may be inadequate to support maximum milk and milk component synthesis. Study objectives were to evaluate whether conjugated linoleic acids- (CLA-) induced milk fat depression (MFD) during heat stress would allow for increased milk and milk component synthesis. In addition, CLA effects on production variables and its ability to induce MFD were compared between Holstein and Brown Swiss cows. Multiparous cows (n = 8, Holstein; n = 5, Brown Swiss) averaging 97 +/- 17 d in milk were used in a crossover design during the summer (mean temperature-humidity index = 75.7). Treatment periods were 21 d with a 7-d adaptation period before and between periods. During adaptation periods, all cows received a supplement of palm fatty acid distillate (242 g/d). Dietary treatment consisted of 250 g/d of CLA supplement (78.9 g/d of CLA) or 242 g/d of palm fatty acid distillate to provide equal amounts of fatty acids. The CLA supplement contained a variety of CLA isomers (3.0% trans-8, cis-10; 3.4% cis-9, trans-11; 4.5% trans-10, cis-12; and 4.8% cis-11, trans-13 CLA). Treatments were applied 2 x/d with half of the supplement top-dressed at 0600 h and the remainder top-dressed at 1800 h. There was no overall treatment effect on dry matter intake (23.9 kg/d), milk yield (40.0 kg/d), somatic cell count (305,000), protein (2.86%), or lactose content (4.51%) or yields of these milk components. Supplementation with CLA decreased overall milk fat content and yield by 26 and 30%, irrespective of breed. The reduction of milk fat content and yield was greatest on d 21 (28 and 37%, respectively). Energy availability predicted by energy balance was improved with CLA supplementation compared with controls (3.7 vs. 7.1 Mcal/d, respectively). Respiration rate (78 breaths/min) and skin temperature (35.4 degrees C) during maximum heat load were not affected by treatment. The group receiving CLA had higher total milk fat CLA concentration (9.3 vs. 4.9 mg/g). Supplementation with CLA induced MFD and altered milk fat composition similarly between breeds and improved calculated energy balance during heat stress, but had no effect on production measures under these conditions. PMID- 15829666 TI - Short communication: Ca2+-adenosine triphosphatase protein expression in the mammary gland of periparturient cows. AB - The objectives of this study were to measure the changes in protein expression of the mammary Ca2+-ATPases during the periparturient period and to determine whether Ca2+-ATPase protein expression in the mammary gland is related to milk fever (MF) development. Abundance of Ca2+-ATPase in mammary tissue and milk fat globule membranes was determined by Western blotting. The secretory pathway Ca2+ ATPase was elevated prepartum in mammary tissue from cows that developed MF compared with non-MF cows. PMID- 15829667 TI - Digestibility in lactating cows of diets containing whole Pima treated with sodium hydroxide versus Akala or Pima cottonseed. AB - This study examines the effect of NaOH treatment on the nutritive value of Pima cottonseed for lactating cows. Treatment of Pima cottonseed with NaOH increased the extent of dry matter (DM) and neutral detergent fiber in vitro digestibility. Three groups of cows were fed 3 similar total mixed rations (TMR) differing only in the type of cottonseed fed, either whole linted Akala, whole unlinted Pima, or whole Pima treated with 4% NaOH (T-Pima). Dry matter and organic matter (OM) intakes of Akala and Pima TMR were similar and tended to be lower compared with cows fed T-Pima. This was reflected in similar intake of crude protein and lipids by cows fed the 3 TMR. Digestibility of DM, OM, and crude protein were similar in cows fed Pima and T-Pima diets, and slightly higher in cows fed the Akala TMR. Lipid digestibility was highest in cows fed Akala TMR, whereas cellulose and NDF digestibility were lowest in Pima TMR. In the 3 TMR, there was a large gap between the high values of "digestibility of intact seeds" and actual in vivo digestion of DM and OM. Similar yields of milk and milk protein were obtained in cows fed the Akala and Pima TMR, whereas higher yields of milk, milk protein, milk fat, and 4% fat-corrected milk were found in the T-Pima cows. Similar efficiency of DM intake for fat-corrected milk production was achieved by cows fed the 3 TMR. PMID- 15829668 TI - Short communication: evaluation of a color method for testing immunoglobulin G concentration in goat colostrum. AB - Colostrum samples (n = 1084) of first and second milking from Majorera goats were taken. The immunoglobulin (Ig) G concentrations estimated by measurement of the color of goat colostrum and by the radial immunodiffusion technique were compared. Least squares analysis of the relationship between the color measurement method and IgG concentration resulted in a significant linear relationship. Using 20 mg of IgG/ mL of colostrum as the cut-off point for colostrum selection, the sensitivity, specificity, and negative predictive value of the color method as a test of IgG concentration in goat colostrum were 93.03, 71.43, and 78.12%, respectively. PMID- 15829669 TI - Effects of flaxseed on protein requirements and N excretion of dairy cows fed diets with two protein concentrations. AB - Thirty-eight midlactating Holstein cows averaging 597 kg of body weight (SD = 59) were used to determine the effects of dietary flaxseed on protein requirement and N excretion in urine and feces. Milk yield and composition, intake, and digestibility were also determined. Cows were allotted from wk 20 to 30 of lactation to 1 of 4 TMR containing 1) no flaxseed (control) and 16% protein (MPC), 2) whole flaxseed and 16% protein (MPF), 3) no flaxseed (control) and 18% protein (HPC), and 4) whole flaxseed and 18% protein (HPF). Cows fed high protein diets had greater feed intake than those fed medium protein diets (20.2 vs. 18.4 kg/d), and cows fed no flaxseed had greater dry matter intake than those fed flaxseed (20.1 vs. 18.5 kg/d). Milk yield was lower for cows fed MPF (20.3 kg/d) than for those fed HPC (24.4 kg/d), HPF (24.9 kg/d), or MPC (24.0 kg/d). Milk protein and lactose concentrations were similar for cows fed MPC and HPC, but flaxseed decreased milk protein concentration in cows fed MPF or HPF compared with cows fed the control diets. Milk fat concentration was similar in cows fed diets with or without flaxseed, but it was decreased by higher protein concentration. Digestibility was generally reduced when diets contained flaxseed and lower protein concentration. Dietary protein had no effect while dietary flaxseed increased fecal N excretion. Retention of N was lower in cows fed flaxseed compared with cows fed the control diets. Feeding flaxseed decreased milk concentrations of short- and medium-chain fatty acids and increased those of long-chain fatty acids. Flaxseed had no effect on the dietary requirement of N by midlactating dairy cows. PMID- 15829670 TI - Dietary manipulation in dairy cattle: laboratory experiments to assess the influence on ammonia emissions. AB - Improvements to the efficiency of dietary nitrogen use by lactating dairy cattle can be made by altering the concentration and form of protein in the diet. This study collected urine and feces from dairy cows from selected crude protein (CP) treatments of 2 lactation studies. In the first trial, collections were made from cattle fed a diet with high (19.4%) or low (13.6%) CP content (HCP and LCP, respectively). In the second trial, collections were made from cattle fed diets in which the forage legume component was alfalfa (ALF) or birdsfoot trefoil with a low (BFTL) or high (BFTH) concentration of condensed tannins (CT). A system of small laboratory chambers was used to measure NH3 emissions over 48 h from applications of equal quantities of urine and feces to cement (simulating a barn floor) and from applications of slurries, made by combining feces and urine in the proportions in which they were excreted for each treatment, to soil. Reducing dietary CP content resulted in less total N excretion and a smaller proportion of the excreted N being present in urine; urine N concentration was 90% greater for HCP than LCP. Surprisingly, NH3 emissions from the barn floor were similar in absolute terms despite the great differences in urine urea-N concentrations, presumably because urease activity was limiting. Cumulative emissions from fresh slurries applied to soil represented 18% of applied N for both HCP and LCP. Following storage at 20 degrees C for 2 wk, cumulative emissions from LCP were much lower than for HCP, representing 9 and 25% of applied N, respectively. Emissions were also lower when expressed as a proportion of slurry total ammoniacal N (TAN) content (24 and 31%, respectively) because of treatment differences in slurry pH. Increasing CT content of the dietary forage legume component resulted in a shift in N excretion from urine to feces. Cumulative NH3 emissions from the barn floor were greater for ALF than for BFTL or BFTH. Emissions from fresh and stored slurries were in proportion to slurry TAN contents, with approximately 35% of applied TAN being lost for all treatments. Emissions expressed as a proportion of total N applied were consistently lower for BFTH than for ALF. PMID- 15829671 TI - Partition of nitrogen excretion in urine and the feces of holstein replacement heifers. AB - Increasing public concern has been focused on animal production systems as a major nonpoint source of pollution. These studies were conducted to further our understanding of whole-animal N metabolism, N excretion, and its partition between feces and urine in growing dairy heifers. Isocaloric diets [2.31 Mcal of metabolizable energy (ME)/kg of dry matter (DM)], ranging from 12.4 to 34.2 g of N/kg of DM, were fed to Holstein heifers in 2 experiments at approximately 1.8 times maintenance. Diets were formulated to provide 54 to 143% of the ruminal ammonia requirements as predicted by the Cornell Net Carbohydrate and Protein System. Increasing the N content of the diet increased urinary N excretion and N balance, but did not affect fecal N excretion. Holstein heifers fed low N diets were able to maintain growth rates consistent with current recommendations while at the same time reducing N excretion, in particular nitrogenous compounds that are readily converted to ammonia. However, more research is needed before this type of diet is recommended for growing heifers because of possible changes in body composition that may affect future milk production and performance. PMID- 15829672 TI - Effects of an enhanced vitamin A intake during the dry period on retinoids, lactoferrin, IGF system, mammary gland epithelial cell apoptosis, and subsequent lactation in dairy cows. AB - Studies in vitro show important interactions among vitamin A, lactoferrin, and insulin-like growth factor (IGF) binding proteins (IGFBP) and, thus, the IGF system. As a consequence, mammary gland epithelial cell proliferation and apoptosis during the bovine dry period and potential milk yield may be affected. We have studied effects of feeding vitamin A (550,000 IU/ d) that exceed daily requirements about 8-fold for up to 2 mo to dairy cows during the dry period on concentrations of retinol and its metabolites in plasma and milk, milk lactoferrin, plasma and milk IGF-I and IGFBP-3, lactoferrin and IGF-I mRNA levels in mammary gland tissue, mammary gland apoptosis, and 100-d milk yield in the ensuing lactation. In the group supplemented with vitamin A, the peripartal decrease of plasma retinol was delayed and attenuated, and colostral retinol plus retinylester concentration was enhanced, but colostral beta-carotene concentration decreased. The retinoic acid isomer 9,13-dicis retinoic acid that coeluted with 13-cis retinoic acid, was the predominant circulating retinoic acid and was higher in GrA than the control group. Plasma IGFBP-3 concentrations were positively correlated with plasma retinol concentrations (r = 0.51), but there were no group differences. Numbers of apoptotic epithelial cells in mammary epithelium were higher at drying off and parturition than in the middle of the dry period, coinciding with high concentrations of IGF-I and lactoferrin in mammary secretions. At parturition, numbers of apoptotic cells in mammary gland biopsies in cows supplemented with vitamin A were higher than in control cows. In conclusion, supplementation of dairy cows during the dry period with high amounts of vitamin A did not significantly modify concentrations of lactoferrin, IGFBP-3, and IGF-I in plasma and in mammary secretions, but slightly decreased energy corrected 100-d milk yield and milk fat yield, possibly because of enhanced apoptic rates of mammary cells. PMID- 15829673 TI - Effect of halofuginone lactate on the occurrence of Cryptosporidium parvum and growth of neonatal dairy calves. AB - Thirty-one Holstein bull calves were purchased at birth from 3 dairy farms in Eastern Ontario. Each calf was assigned at random to oral treatment with either 5 mg of halofuginone lactate in 10.0 mL of aqueous carrier solution (Halocur, base comprised 10 mg of benzoic acid, 100 mg of lactic acid, and 0.3 mg of tartrazine) or 10 mL of placebo (Halocur base minus the active ingredient, halofuginone lactate) administered 15 to 30 min after morning milk feeding for the first 7 d of life. Intakes of milk, calf starter, and water, and fecal consistency score were recorded daily for 56 d. Calf weights were recorded weekly for 56 d. Fecal samples were taken from all calves at approximately 2, 7, 14, 21, and 28 d of age for isolation of Cryptosporidium parvum oocysts. Logistic and linear regression analyses were used to assess the effect of treatment on the incidence of diarrhea and C. parvum infection status. The odds of C. parvum shedding among calves in the halofuginone lactate-treated group was 70% lower than the odds of shedding among calves in the placebo group. In calves treated with halofuginone lactate, no oocyst shedding occurred until 2 wk of age, whereas 12.5% of calves in the placebo group began shedding oocysts during wk 1. From all ages of placebo treated calves, 31 of 73 samples (42.5%) were positive for C. parvum, whereas only 15 of 67 samples (22.4%) from all ages of halofuginone lactate-treated calves tested positive. The largest number of C. parvum-positive samples occurred in the third week of life. There was a significant delay of 3.1 d in the incidence of diarrhea among calves treated with halofuginone lactate. Intake of milk and starter, body weight gains, and age at weaning were not significantly different between treatment groups. PMID- 15829674 TI - Digestion of feed amino acids in the rumen and intestine of steers measured using a mobile nylon bag technique. AB - The disappearance of dry matter (DM), crude protein (CP), and amino acids (AA) in steers after rumen incubation and intestinal passage of alfalfa hay, barley hay, corn silage, barley grain, corn grain, wheat bran, meat meal, fish meal, cottonseed meal, and soybean meal were measured in 3 steers using a mobile nylon bag technique. Ruminal degradation of individual AA differed between feedstuffs. For barley hay and corn silage, the ruminal disappearance of total AA was higher and lower than the other feedstuffs, respectively. The intestinal digestibility of total AA in alfalfa hay was lower than the digestion of CP. The intestinal digestibility of Arg and His was higher than that of total AA in alfalfa hay, meat meal, cottonseed meal, soybean meal, barley hay, and wheat bran. In addition, the intestinal digestibility of Lys was higher than that of total AA in alfalfa hay, meat meal, cottonseed meal, soybean meal, barley hay, corn silage, and wheat bran. The intestinal disappearance of CP in most cases was higher than that of DM. The results indicated that feedstuffs with lower ruminal disappearance of DM, CP, total AA, essential AA, and nonessential AA generally had a higher intestinal disappearance, resulting in a relatively constant total tract disappearance. These results could be used to improve the current system of diet formulation in ruminants. PMID- 15829675 TI - Aerobic stability of wheat and orchardgrass round-bale silages during winter. AB - Using recently developed technology, balage is often stored in large (1.2 x 1.2 m) round bales that are wrapped in plastic film with an in-line wrapper. The aerobic stability of this fermented forage is important, particularly during winter months when it is fed to livestock or sold as a cash crop. Two types of forage, orchardgrass [Dactylis glomerata L.; 54.4% dry matter (DM)] and wheat (Triticum aestivum L.; 62.4% DM), were packaged in large round bales and wrapped with an in-line wrapper during May 2002. Twenty-one bales of each balage type were unwrapped and exposed to air on Dec. 10, 2002 for 0, 2, 4, 8, 16, 24, or 32 d (ambient temperature range = 0.6 to 19.4 degrees C) to evaluate aerobic stability. For both orchardgrass and wheat balage, final bale weight, concentration of DM, and pH were not affected by exposure time. Across both balage types, DM recoveries were > or = 97% for all bales, indicating that both balage types were very stable when exposed to air. For orchardgrass balage, exposure time had no effect on concentrations of NDF, ADF, hemicellulose, cellulose, or lignin, thereby indicating that little deterioration occurred. Similarly, no contrast relating any fiber component with exposure time was significant for wheat balage. Concentrations of crude protein (CP) were not affected by exposure time for wheat balage, but there was a tendency for exposed orchardgrass bales to have greater concentrations of CP than bales sampled on d 0. Exposure time had no effect on 48-h in situ digestibility of DM for wheat balage, but there was a tendency for a linear increase with exposure time for orchardgrass balage. However, the overall range (78.2 to 80.5%) over the 32-d exposure period was very narrow, and this response is probably of limited biological significance. Generally, concentrations of fermentation acids were low, primarily because of the high concentration of DM within these balages, and only minimal changes in these acids were observed over the exposure interval. These results suggest that the balage evaluated in this trial during winter conditions was very stable after exposure to air for up to 32 d. This should allow for considerable flexibility with respect to feeding, transport, and marketing of balage during winter months without significant aerobic deterioration. PMID- 15829676 TI - Comparison of techniques for estimating herbage intake of grazing dairy cows. AB - For estimating herbage intake during grazing, the traditional sward cutting technique was compared in grazing experiments in 2002 and 2003 with the recently developed n-alkanes technique and with the net energy method. The first method estimates herbage intake by the difference between the herbage mass before and after grazing and the regrowth between the 2 points in time. The second technique estimates herbage intake by the ratio of a dosed even-chain synthetic n-alkane (C32) and a naturally occurring odd-chain n-alkane (C31 or C33) in the herbage and feces. The third technique calculated the intake from the animal's energy requirements for milk production and maintenance. The sward cutting technique estimated herbage intake with the highest coefficient of variation and had different results in the 2 experimental years. The n-alkanes method yielded less variable results, whereas the net energy method gave the least variable results. In 2002, the estimates of the alkane ratio C32:C33 were best related with estimations of the net energy method. In 2003, the estimates of the alkane ratio C32:C31 were best related. The estimate based on the alkane ratio C32:C33 had a lower coefficient of variation than the one based on the alkane ratio C32:C31. Therefore, the C32:C33 alkane method was considered to be a better direct estimator for herbage intake by grazing lactating dairy cows. PMID- 15829677 TI - Consumer acceptability of conjugated linoleic acid-enriched milk and cheddar cheese from cows grazing on pasture. AB - Two experiments were conducted to study the consumer acceptability attributes of conjugated linoleic acid (CLA)-enriched milk and cheese from cows grazing on pasture. In experiment 1, 15 cows were fed either a diet containing 51% alfalfa hay plus corn silage and 49% concentrate [total mixed ration (TMR)], were grazed on pasture, or were grazed on pasture and received 3.2 kg/d of a grain mix. The grain mix contained 75% full-fat extruded soybeans (FFES), 10% corn, 10% beet pulp, and 5% molasses. During the final 3 wk of the 6-wk experiment, milk was evaluated for sensory attributes. In experiment 2, 18 cows were fed similar diets as in experiment 1, except replacing the group of cows grazed on pasture and receiving the grain mix was a group of cows grazed on pasture and receiving 2.5 kg/d per cow of the FFES; Cheddar cheese was manufactured from milk. Average CLA contents (g/100 g of fatty acid methyl esters) were 0.52, 1.63, and 1.69 in milk and 0.47, 1.47, and 1.46 in cheese from cows fed a TMR, grazed on pasture, and grazed on pasture and fed the grain mix, respectively. An open and trained panel evaluated CLA-enriched milk for mouth-feel, color, flavor, and quality and evaluated cheese for color, flavor, texture, and quality. Open and trained panel evaluations of milk and cheese showed no differences among treatments for any of the attributes, except that the trained panel detected a more barny flavor in milk from cows grazing pasture compared with milk from cows fed the TMR only. Results suggest that consumer acceptability attributes of CLA-enriched milk and cheese from cows grazing pasture is similar to those of milk and cheese with low levels of CLA. PMID- 15829678 TI - Within-day feeding behavior of lactating dairy cows measured using a real-time control system. AB - The suitability of a statistical model for describing within-day feeding behavior, and potential relationships between model parameters and commonly measured experimental variables were examined. Forty multiparous, midlactation Holstein cows were fed using a real-time control system to record the date of each visit to a feeder, entrance time, exit time, and feed consumed over a 6-wk period. Daily feed consumption, number of visits, meal duration, and within-visit rate of food intake were then calculated. Two peaks in within-day rates of feed intake were indicated, suggesting that feeding activity was randomly distributed around each peak, that is, binormal. Parameters describing the distributions (means, standard deviations, and the percentage of total feeding activity associated with each peak) were estimated. An adjusted average of 91% of the variation in within-day feeding activity was explained by the binormal model. Relationships between model parameters and commonly measured experimental variables were also identified; behavioral traits were correlated with total feed intake. Feeding activity patterns in literature data were also amenable to reanalysis by the binormal model. Lactating cows consistently exhibited a distinct diurnal pattern in feeding activity; they were most active near sunrise and again near sunset (crepuscular). Effects of various management operations (e.g., feeding and milking times and frequencies, and lighting) on within-day feeding patterns remain to be established, although a statistical model for evaluating them is now available. The patterns may have important implications for scheduling management activities to maximize feed intake and production. PMID- 15829679 TI - Effect of heat stress on production of Mediterranean dairy sheep. AB - A study on heat stress in Mediterranean dairy sheep was undertaken with the objective to examine the relationship between milk production and heat stress, to estimate the additive genetic variances of milk production traits and heat tolerance, and to investigate the possibility of future selection for increased heat tolerance. Production data included 59,661 test-day records belonging to 6624 lactations of 4428 lactating ewes from 17 flocks collected from 1994 through 2003. The traits investigated were daily milk yield, fat and protein percentage, and daily yield of fat-plus-protein. The pedigree file consisted of 5306 animals; in addition to the 4428 animals with records, 188 male and 690 female ancestors were included. Heat stress was modeled by using data from a weather station. Apart from the effects of the weather conditions of the milk recording test-day, the effects of the preceding 1, 2, and 3 d were determined. Because longer periods of heat stress might have a more severe effect than shorter periods, 2-, 3-, and 4-d periods were also considered, by averaging the weather data measurements. Fixed regression analyses were based on models that included effects of flock nested within year of test-day, DIM (days in milk) class x parity class, and several types of weather indicators. The preferred model using the temperature-humidity index (THI) gave a smoother pattern than did the model with temperature x humidity interaction. Both daily milk and fat-plus-protein yield appeared to decrease at THI > or = 23, in all periods considered. Based on the 4-d period, yield decreased for each unit increase of THI above 23 [-62.8 g/unit (-4.2%) for daily milk yield and -8.9 g/unit (-4.9%) for daily fat-plus protein yield]. Fat and protein percentages appeared to be unaffected by heat stress. A test-day repeatability model was applied for estimation of genetic parameters. The genetic correlations between the general additive effect and the additive effect of heat tolerance were negative (approximately -0.8) for both daily milk and fat-plus-protein yields in all periods considered. Therefore, milk yield is antagonistic with heat tolerance, and selection only for increased milk production will reduce heat tolerance. PMID- 15829680 TI - Inbreeding in Danish dairy cattle breeds. AB - The purpose of this study was to monitor current and predict future rates of inbreeding in the Danish dairy breeds. Calves born from 1999 until 2003 and registered as Danish Holstein (1,883,983), Danish Jersey (336,966), or Danish Red (261,047) were reference populations. Average complete generation equivalent was approximately 7. For calves born in 2003, average inbreeding was 3.9, 3.4, and 1.4% for Holstein, Jersey, and Danish Red, respectively. In recent years, effective population sizes were 49, 53, and 47, respectively. Based on coancestry statistics, future effective population sizes will be 43, 42, and 51, respectively. The effective number of founders, effective number of ancestors, and effective number of founder genomes were calculated. These measures of genetic diversity were all low for Holstein and Jersey and somewhat larger for Danish Red. The most important ancestors of Danish Holstein were Elevation (13.8%), Chief (10.9%), and Bell (8.5%). The most important ancestor of Danish Red was Momentum (9.4%), a Red Holstein-Friesian. The most important ancestor for Danish Jersey was FYN Lemvig (12.1%) with a large number of progeny in the reference population. The results of this study indicate the necessity for active management of the rate of inbreeding in the future. PMID- 15829681 TI - Accounting for pregnancy diagnosis in predicting days open. AB - The system for estimating days open for cows with no subsequent lactation was examined to determine if estimates should vary depending on pregnancy diagnosis. Pregnancy diagnosis information was unavailable when the original prediction system was developed, but collection was begun in 2002. New prediction equations were estimated from nearly 1.1 million Holstein lactations for 20-d intervals from 110 to 250 days in milk (DIM). Use of pregnancy diagnosis improved accuracy compared with the original system. The improvement was particularly evident for lactations of cows confirmed to be open in the 130-to-149 DIM interval, where predicted days open increased by > 96 d. For lactations of cows with a confirmed pregnancy, predicted days open decreased by 18 d for the same interval. Prediction errors decreased with increasing DIM. Jersey lactations averaged fewer days open, but in most cases Holstein solutions provided adequate predictions. Specific adjustments were generated for Jersey lactations with no breedings reported. Those adjustments reduced the predicted days open averaged across parity by an amount that increased from 9 to 27 d with DIM interval. The new prediction equations were implemented for November 2004 evaluations for daughter pregnancy rate. PMID- 15829682 TI - Short communication: predominance of beta-casein (CSN2) C allele in goat breeds reared in Italy. AB - A protocol for the rapid and simultaneous genotyping of A, C, and 0 'CSN2 alleles in goat was developed by single strand conformational polymorphism polymerase chain reaction (SSCP-PCR) technique. Screening the CSN2 variability in 7 goat breeds reared in Italy validated the genotyping test. The SSCP-PCR technique was also suitable for monitoring CSN2 polymorphism. In particular, the discrimination between CSN2*A and CSN2*C is important because the 2 corresponding protein variants cannot be separated by standard typing techniques. The monitoring of CSN2 variability in the goat breeds indicates the predominance of the C allele. In most breeds, CSN2*C occurred with the highest frequency, except in Saanen where CSN2*A and CSN2*C showed similar frequencies. Variant CSN2*C occurred with a frequency of 0.68 (Camosciata), 0.70 (Jonica), 0.71 (Garganica), 0.82 (Maltese), 0.87 (Cilentana), and 0.97 (Orobica). The alignment among the mature CSN2 sequences of different species suggests that CSN2*A is the ancestral allele compared with CSN2*C. Interestingly, the CSN2*A goat variant showed higher frequencies in selected breeds (Saanen and Camosciata). PMID- 15829683 TI - Derivation of sustainable breeding goals for dairy cattle using selection index theory. AB - The objective was to present 2 methods for the derivation of nonmarket values for functional traits in dairy cattle using deterministic simulation and selection index theory. A nonmarket value can be a value representing animal welfare and societal influences for animal production, which can be added to market economic values in the breeding goal to define sustainable breeding goals. The first method was restricted indices. A consequence of adding a nonmarket value to a market economic value for a given functional trait is less selection emphasis on milk yield. In the second method, the loss in selection response in milk resulting from greater emphasis on functional traits was quantified. The 2 methods were demonstrated using a breeding goal for dairy cattle with 4 traits (milk yield, mastitis resistance, conception rate, and stillbirth). Nonmarket values derived separately using restricted indices were 0.4 and 2.6 times the value of market economic values for mastitis resistance and conception rate, respectively. Nonmarket values for mastitis resistance and conception rate were both lower when derived simultaneously than when derived separately. This was due to the positive genetic correlation between mastitis resistance and conception rate, and because both traits are negatively correlated with milk yield. Using the second method and accepting a 5% loss in selection response for milk yield, nonmarket values for mastitis, conception rate, and stillbirth were 0.3, 1.4, and 2.9 times the market economic values. It was concluded that the 2 methods could be used to derive nonmarket values for functional traits in dairy cattle. PMID- 15829684 TI - A microfiltration process to maximize removal of serum proteins from skim milk before cheese making. AB - Microfiltration (MF) is a membrane process that can separate casein micelles from milk serum proteins (SP), mainly beta-lactoglobulin and alpha-lactalbumin. Our objective was to develop a multistage MF process to remove a high percentage of SP from skim milk while producing a low concentration factor retentate from microfiltration (RMF) with concentrations of soluble minerals, nonprotein nitrogen (NPN), and lactose similar to the original skim milk. The RMF could be blended with cream to standardize milk for traditional Cheddar cheese making. Permeate from ultrafiltration (PUF) obtained from the ultrafiltration (UF) of permeate from MF (PMF) of skim milk was successfully used as a diafiltrant to remove SP from skim milk before cheese making, while maintaining the concentration of lactose, NPN, and nonmicellar calcium. About 95% of the SP originally in skim milk was removed by combining one 3 x MF stage and two 3 x PUF diafiltration stages. The final 3 x RMF can be diluted with PUF to the desired concentration of casein for traditional cheese making. The PMF from the skim milk was concentrated in a UF system to yield an SP concentrate with protein content similar to a whey protein concentrate, but without residuals from cheese making (i.e., rennet, culture, color, and lactic acid) that can produce undesirable functional and sensory characteristics in whey products. Additional processing steps to this 3-stage MF process for SP removal are discussed to produce an MF skim retentate for a continuous cottage cheese manufacturing process. PMID- 15829685 TI - Impacts of the Doha Round framework agreements on dairy policies. AB - Dairy is highly regulated in many countries for several reasons. Perishability, seasonal imbalances, and inelastic supply and demand for milk can cause inherent market instability. Milk buyers typically have had more market power than dairy farmers. Comparative production advantages in some countries have led to regulations and policies to protect local dairy farmers by maintaining domestic prices higher than world prices. A worldwide consensus on reduction of border measures for protecting dairy products is unlikely, and dairy will probably be an exception in ongoing World Trade Organization (WTO) negotiations. Under the Doha Round framework agreements, countries may name some products such as dairy as "sensitive," thereby excluding them from further reforms. However, new Doha Round framework agreements depart from the current WTO rule and call for product specific spending caps. Such caps will greatly affect the dairy sector because dairy accounts for much of the aggregate measure of support (AMS) in several countries, including the United States and Canada. Also, the amounts of dairy AMS in several countries may be recalculated relative to an international reference price. In addition, all export subsidies are targeted for elimination in the Doha Round, including export credit programs and state trading enterprises, which will limit options for disposing of surplus dairy products in foreign markets. Currently, with higher domestic prices, measures for cutting or disposing of surpluses have been used in many countries. Supply control, which is not regulated by WTO rules, remains as an option. Although explicit export subsidies are restricted by WTO rules, many countries use esoteric measures to promote dairy exports. If countries agree to eliminate "consumer financed" export subsidies using a theoretical definition and measurements proposed herein as Export Subsidy Equivalents (ESE), dairy exports in many countries may be affected. Although domestic supports and export subsidies will be reduced in the Doha Round, possible exclusion of "sensitive" products from tariff reduction will help some countries' dairy sectors survive after those final agreements. A key concern for those countries will be the simultaneous restriction of surplus disposing measures. With fewer marketing options for surpluses, countries that continue border protection and high internal prices will likely be forced to use domestic supply control programs in the future. PMID- 15829686 TI - Expert assessment study of milking and hygiene practices characterizing very low somatic cell score herds in France. AB - French dairy herds were selected on a national basis through the "Zero Mastitis Objective" Program (ZMP) to display hygiene and milking practices characterizing very low somatic cell score (SCS) herds. The herds selected were stratified in 2 groups. The first group (LOW) included 187 herds within the first 5 percentiles of herds regarding mean SCS for the 36 mo preceding ZMP (36-mo SCS). The second group (MED) included 117 herds within the 50 to 55th percentile of herds regarding 36-mo SCS. Mean milk SCS was 3.09 in the MED herds vs. 1.99 in the LOW herds, which corresponded to 265,000 and 135,000 cells/mL respectively. Moreover, LOW and MED herds did not change from one SCS category to another during ZMP. Potentially explanatory variables, collected by formally trained dairy management experts through observations from attendance at milking and farm visits, were analyzed using multistep logistic regression models. According to final model and expert observations, the probability for a herd to belong to the LOW group was maximized when: 1) winter cleanliness of dry cow shed was good; 2) use of teat spraying was carried out; and 3) California Mastitis Tests were performed at milking. Moreover, the herd probability of belonging to the MED group was maximized when: 1) air admission at teat cup attachment was observed during milking; 2) winter cleanliness of dry cow shed was poor; and 3) the milker spent time during milking to feed calves. Finally, the study highlighted milking and hygiene variables and attitudes appearing as key practices to control herd SCS through precise and safe milking and more attention paid to individual animals and cleanliness of dry cow shed. PMID- 15829687 TI - An overview of radiographic film holders. AB - OBJECTIVES: To provide an overview of radiographic film holders, from their inception in the early 1900s to present day. METHODS: A Medline literature search was conducted for dental studies that used keywords that pertain to radiographic film holders. Hand searches of the bibliographies were also conducted. Sixty articles (dating from 1896 to 1998) that describe new or improved film holders are reviewed here. RESULTS: From the earliest days of dental radiography, dentists attempted to standardize radiographic images and techniques. The focus of researchers in the 1950s to the 1970s was to develop a film holder that would hold the film and allow easy and predictable alignment of the X-ray tube. As research projects became more dependent on dental radiographic measurements, the focus shifted to producing reproducible radiographic images, from which highly repeatable measurements could be made. CONCLUSION: Existing devices have strengths and weaknesses. Readily available devices are adequate for routine clinical use; however, user-friendly and patient-friendly film-holding devices that result in highly reliable and accurate measurements have yet to be introduced. PMID- 15829688 TI - Detection of foreign bodies of the head with digital volume tomography. AB - OBJECTIVES: Foreign bodies occur as a sequela of trauma or therapeutic interventions. If the risks are not too high, therapy of choice is the removal of the foreign body. This is only possible if the existence of a foreign body is detected and it is localized accurately. METHODS: The suitability of digital volume tomography (DVT) for the detection and localization of foreign bodies was investigated. Samples of various materials that are typically found as foreign bodies in the head and neck region were used. RESULTS: All materials investigated are principally detectable with DVT. Highly radiopaque material was detectable down to small sizes. The detectability of less radiopaque substances like wood and resins was poor when located in soft tissue. CONCLUSION: DVT is an appropriate tool for the detection of radiopaque foreign bodies. However, foreign bodies made of wood and resin, although detectable by DVT, can be missed when located in muscular tissue. The detectability decreases further when the foreign body is located adjacent to a highly radiopaque tissue like bone. This has to be taken into consideration when using DVT for foreign body detection. PMID- 15829689 TI - Ultrasound crack detection in a simulated human tooth. AB - OBJECTIVE: Currently, diagnosis of cracked teeth generally depends upon the overall clinical assessment, or on exclusion of other clinical possibilities, not primarily on the direct identification of cracks themselves. Owing to its short wavelength in hard tissues and associated high resolution, ultrasound has the potential to allow detection of cracks within tooth structure. However, ultrasound detection of dental cracks has not previously been achieved. The purpose was to determine if an ultrasound imaging system was capable of imaging cracks in simulated tooth structure. METHODS: A complete ultrasound system including a novel transducer made of PLZT-98, a novel gallium-indium alloy coupling agent, and customized electronic and digital signal processing (DSP) algorithms was developed for the specific application of optimizing crack detection within teeth. A simulated tooth with a known and uniform internal structure and acoustic properties similar to those of natural enamel and dentin was designed to model a human tooth with a crack located in dentin deep to the dentino-enamel junction (DEJ). The distance between the DEJ and a crack of the simulated tooth were calculated. RESULTS: The system unequivocally distinguished between areas with and without a simulated crack. CONCLUSION: A unique ultrasound dental crack detection system using a novel transducer; a novel coupling agent; and customized electronic and digital signal processing (DSP) algorithms has been validated in a simulated tooth. PMID- 15829690 TI - CT findings as a significant predictive factor for the curability of mandibular osteomyelitis: multivariate analysis. AB - OBJECTIVES: To re-evaluate computed tomographic (CT) imaging as a diagnostic tool for mandibular osteomyelitis and to assess the clinical significance of CT findings. METHODS: CT images of 78 patients with mandibular osteomyelitis were reviewed. All patients were classified as cured or non-cured. Each CT finding was investigated for frequency, correlation with duration and disease cure. RESULTS: Of the 78 patients, 49 (63%) were classified as "cured" and 29 (37%) as "non cured". Non-cured had experienced a significantly longer duration of symptoms. The most frequent CT finding was sclerosis and defect in the trabecular bone. Changes of bone width and thickening of the cortical plate were accompanied with longer disease duration. The extent of the diseased area was linearly correlated with the duration of symptoms. The significant factors to discriminate non-cured from cured were the extent of the disease, the number of findings, changes in the bone width, osteosclerosis and thickening of the cortex. Multivariate logistic regression analysis indicated that the extent of the disease and presence of changes in bone width were significant variables correlating with the cure of osteomyelitis. CONCLUSIONS: The extent of disease and the presence of change in bone width shown on CT were significantly correlated with the curability of osteomyelitis. These results indicated the usefulness and importance of CT examination for the diagnosis of mandibular osteomyelitis. PMID- 15829691 TI - The utility of panoramic radiography in temporomandibular joint assessment. AB - OBJECTIVES: Panoramic radiography was used to determine (1) intrarater and inter rater reliability in assessing temporomandibular joint (TMJ) condylar morphology; (2) alteration in condylar shape in patients with temporomandibular disorders (TMD) and controls when matched by age, gender, and state of dentition; and (3) prevalence of condylar abnormalities in individuals with and without TMD. METHODS: One hundred panoramic radiographs were randomly selected from a hospital clinic (45 TMD and 55 non-TMD patients). The images were cropped to include only the temporomandibular apparatus and were independently evaluated by three examiners without knowledge of the patient's clinical status. Multiple statistical tests were performed to evaluate the accumulated data. RESULTS: Intrarater reliability demonstrated substantial agreement, while inter-rater reliability was fair. There was no difference in condylar morphology between patient groups, but mild condylar change was prevalent in all age groups, regardless of TMD status. CONCLUSIONS: Morphological condylar abnormalities are present on panoramic images in all adult age ranges, regardless of status of the dentition or presence of TMD. Condylar shape alone is not an indicator of TMD, and minor condylar discrepancies may have no significance in TMD. PMID- 15829692 TI - Clinical evaluation of the effects of radiographic distortion on the position and classification of mandibular third molars. AB - OBJECTIVE: Panoramic radiographs are routinely employed for surgical planning of unerupted third molars. The objective of this study was to evaluate distortions in the position of mandibular third molars on panoramic radiographs. MATERIALS AND METHODS: Panoramic radiographs of 14 patients with indication for extraction of the mandibular third molars were used. Transurgical impressions of the crowns of the teeth to be extracted and adjacent second molars (n=19) were achieved with addition silicone and poured with type IV plaster for comparison of the inclinations of the third molars on the radiographs and study casts. RESULTS: There was a mean difference of -5.37 degrees between the position of the third molar on the panoramic radiograph and on the study cast (standard deviation 1.46, P<0.05, Student's t-test). CONCLUSION: There is distortion in the position of teeth on the panoramic radiograph, which may influence the surgical planning; however, this does not invalidate it as the main tool for diagnosis and surgical planning of mandibular third molars. PMID- 15829693 TI - CT findings of head and neck anomalies in lacrimo-auriculo-dento-digital (LADD) syndrome. AB - We present computed tomography (CT) findings of a boy with lacrimo-auriculo-dento digital (LADD) syndrome. Bilateral lacrimal glands were not detected and CT imaging of this abnormality in LADD syndrome has not been reported before. Despite suggestions from previous reports of multiple temporal bone abnormalities resulting in hearing losses in these patients, only minor temporal bone abnormalities were seen in our deaf-mute patient. Dysplasia of salivary glands was noted. CT findings of head and neck abnormalities in LADD syndrome are discussed. PMID- 15829694 TI - Longitudinal MRI follow-up of temporomandibular joint internal derangement with closed lock after successful disk reduction with mandibular manipulation. AB - We present a case of bilateral temporomandibular joint (TMJ) internal derangement with closed lock, which was followed over 11 years using serial MRI after successful disk reduction with mandibular manipulation. MRI of the bilateral TMJs before and after mandibular manipulation followed by anterior repositioning splint showed that the disk improved from non-reducible to reducible, and that the reducing disk persisted for 11 years, although minor changes in configuration and position of the disk occurred. These MRI findings suggest that non-surgical treatment with successful mandibular manipulation prevents progressive changes in the TMJ. This case report complements observations regarding the longitudinal course of TMJ internal derangement with closed lock after non-surgical treatment. PMID- 15829695 TI - Primary intraosseous carcinoma of the mandible. AB - Primary intraosseous carcinoma (PIOC) is a rare lesion in the jawbone. We report a case of de novo PIOC of the mandible, first seen by the family dentist. Our case highlights that radiographic examination is one of the most effective methods for detecting PIOCs. PMID- 15829696 TI - Collagenous fibroma (desmoplastic fibroblastoma): a new case originating in the palate. AB - A 49-year-old woman presented with a painless slowly growing but huge polypoid palatal mass. On magnetic resonance imaging (MRI), areas of low signal intensity were consistent with a fibrous tumour. T(1) and T(2) weighted MRI after Gd-DTPA administration highlighted a well-circumscribed, purely soft tissue process. Histological analysis of the operative specimen detected collagenous fibroma (desmoplastic fibroblastoma). The patient remains disease-free 4 years after excision. PMID- 15829697 TI - Central haemangioma: variance in radiographic appearance. AB - Central haemangioma, a "great mimicker" which, fortunately, is a relatively rare condition, may pose a lethal risk for the patient. The diagnosis may become apparent only during biopsy or tooth extraction, which poses a risk of lethal exsanguination; therefore a correct diagnosis is desirable before any biopsy is undertaken. The clinician may not anticipate the severe haemorrhage because of vague clinical history, physical findings and ambiguous radiographic characteristics of the lesion. We report a case of central haemangioma of the mandible whose clinical and radiographic features were equivocal. In addition, an attempt is made to discuss all possible radiographic presentations of central haemangioma and consider differential diagnosis. This case is significant for the reason that it had diverse radiographic appearances in various areas of the lesion in different projections. PMID- 15829698 TI - Metastatic retinoblastoma of the maxilla and mandible. AB - Metastatic retinoblastoma of the jaws is very rare. We present a 4-year-old boy with metastatic retinoblastoma that involved both the maxilla and mandible simultaneously. Enhanced CT indicated bone-destructive masses with partially non enhanced area and enhanced margin in the right maxilla and left mandible. MRI showed well-delineated masses that were isointense on T(1) weighted images and hyperintense on T(2) weighted images. Four weeks after chemotherapy and bone marrow transplantation, the size of lesions remarkably decreased. The patient died 19 months later with extensive tumour metastases despite additional chemotherapy. In this case, the dental crypt of a permanent tooth was considered the potential target through which retinoblastoma metastasized to the jaws. PMID- 15829700 TI - Modulation of peroxisome proliferator-activated receptor delta activity affects neural cell adhesion molecule and polysialyltransferase ST8SiaIV induction by teratogenic valproic acid analogs in F9 cell differentiation. AB - It has been suggested that the teratogenic effects of the antiepileptic drug valproic acid (VPA) is reflected in vitro by the differentiation of F9 cells, activation of peroxisome proliferator-activated receptor delta (PPARdelta), and inhibition of histone deacetylases (HDACs). The aim of this study was to identify genes involved in the differentiation of F9 cells induced by VPA, teratogenic VPA derivatives, or the HDAC inhibitor trichostatin A (TSA) and to characterize the role of PPARdelta. Treatment of the cells with teratogenic VPA derivatives or TSA induced differentiation of F9 cells, mRNA, and protein expression of the neural cell adhesion molecule (NCAM) as well as activated the 5'-flanking region of the NCAM promoter, whereas nonteratogenic VPA derivatives had no effect at all. The polysialyltransferases [ST8SiaIV (PST1) and ST8SiaII] are responsible for the addition of polysialic acid (PSA) to NCAM. The mRNA expression of PST1 was highly induced by only teratogenic VPA derivatives and TSA. As shown by fluorescence activated cell sorting analysis the level of PSA was higher after treatment of F9 cells with teratogenic VPA derivatives. It is interesting that overexpression of the PPARdelta but not PPARalpha or PPARgamma in F9 cells resulted in higher induction of NCAM mRNA and protein expression and of PST1 mRNA expression (and a higher PSA level) than in mock-transfected F9 cells. Furthermore, repression of PPARdelta activity in F9 cells inhibited these effects. We conclude that NCAM and PST1 are molecular markers in F9 cell differentiation caused by treatment with teratogenic VPA compounds or TSA and suggest that in addition to HDAC inhibition PPARdelta is involved in the signaling pathway. PMID- 15829701 TI - Suppressors of cytokine signaling regulate angiotensin II-activated Janus kinase signal transducers and activators of transcription pathway in renal cells. AB - Suppressors of cytokine signaling (SOCS) family is constituted by cytokine inducible proteins that modulate receptor signal transduction via tyrosine kinases, mainly the Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathway. Differential SOCS expression was noted in renal cells that were incubated with inflammatory stimuli, but the role of SOCS in the pathogenesis of renal diseases is not yet well defined. Because angiotensin II (Ang II) plays a key role in renal disease, SOCS proteins were studied as a novel mechanism involved in the negative regulation of Ang II-mediated processes. Systemic Ang II infusion for 3 d increased the renal mRNA expression of SOCS-3 and SOCS-1. SOCS protein synthesis was found in glomerular mesangial area and tubules. In cultured mesangial cells and tubular epithelial cells, Ang II induced a rapid and transient SOCS-3 and SOCS-1 expression in parallel with JAK2 and STAT1 activation. In both cell types, overexpression of SOCS proteins prevented the STAT activation in response to Ang II. SOCS expression observed in Ang II infused rats and in Ang II-stimulated cells was significantly inhibited by treatment with AT(1) but not AT(2) receptor antagonist and was attenuated in mesangial cells from AT(1a)-deficient mice, demonstrating the implication of AT(1) in those responses. In SOCS-3 knockdown studies, antisense oligonucleotides inhibited the expression of SOCS-3 and increased the Ang II-induced STAT activation and c-Fos/c-Jun expression, then resulting in a more severe renal damage. These results suggest that SOCS proteins may act as negative regulators of Ang II signaling in renal cells and implicate SOCS as important modulators of renal damage. PMID- 15829702 TI - Pathochemical toxicity of perfluorocarbon-5070, a liquid test performance fluid previously used in dialyzer manufacturing, confirmed in animal experiment. AB - In the light of clustered deaths in late 2001 associated with hemodialysis (HD), this article analyzes the pathochemical toxicity of the perfluorocarbon-5070 (PF 5070), a liquid used as test performance fluid for detecting capillary leaks during dialyzer manufacturing. Residual PF-5070 in some Athane dialyzers of the involved brands was infused in the injured patients during hemodialysis. The clinical presentation was in contrast with other previously described severe reactions to HD. Foam material was discovered in the right ventricle and caval vein of the patients who underwent postmortem examination. Deaths were attributed to gas embolism without the external causes identified. To explore the pathochemical toxicity of the inert liquid PF-5070, an animal model was developed. In a rabbit model, single slug intravenous injections as bolus of increasing doses of PF-5070 were performed. In a first set of experiments, three groups of three rabbits were administered increasing doses of PF-5070 at 4, 40, or 160 microl/kg. After intravenous injection, the animals were observed for clinical signs of adverse effects and underwent autopsy after death. Doses were normalized to animal body weight to allow comparison with supposed patient exposure. Five of nine rabbits died soon after PF-5070 dosing: One rabbit died within 4 h in the 4 microl/kg group, one rabbit died within 30 min in the 40 microl/kg group, and three rabbits died within 30 min in the 160 mul/kg group. In a second set of experiments, six rabbits were injected with a lethal dose of PF 5070 to analyze clinical symptoms and pathophysiology. All rabbits died on the day of dosing and displayed neurologic disorders (paralysis, nystagmus, rigidity, convulsions), then breathing abnormalities (rapid breathing, salivation, dark mucous membrane), and fatal collapse. Autopsy of rabbits showed evidence of gas retention in the lung tissue and gas bubbles in the right cardiac cavities. Histologic findings included alveolar hemorrhage with pulmonary edema, cerebellum, and cortex patchy areas of infarction. Single-dose intravenous administration of PF-5070 reproduced in a rabbit model the pathophysiologic effects observed in the hemodialysis patients. Severity of the symptoms observed in the animals was dose-dependent. Clinical and pathologic findings can be explained by the capacity of perfluorocarbon to emulsify blood at body temperature, to increase partial pressure in the pulmonary capillary bed, and to form bubbles in the pulmonary capillary circulation, thus blocking lung and visceral perfusion. Such experimental findings indicate the toxicity of PF-5070 administered intravenously and make the pathochemical toxicity link with the hemodialysis-related deaths caused by the presence of residues of PF-5070 in the Althane dialyzers. We conclude, in light of this outbreak and the subsequent investigations, that liquid PF-5070 is a highly toxic compound when administered intravenously because of its emulsifying properties. The use of PF-5070 or any liquid fluorocarbon compounds in medical devices with blood contact and particularly in the dialyzer manufacturing should be considered with caution. PMID- 15829703 TI - Individual PKC-phosphorylation sites in organic cation transporter 1 determine substrate selectivity and transport regulation. AB - To elucidate the molecular mechanisms underlying stimulation of rat organic cation transporter type 1 (rOCT1) by protein kinase C (PKC) activation, functional properties and regulation of rOCT1 stably expressed in HEK293 cells after site-directed mutagenesis of putative PKC phosphorylation-sites were compared with wild-type (WT) rOCT1 using microfluorometric measurements with the fluorescence organic cation 4-(4-(dimethylamino)styryl)-N-methylpyridinium (ASP(+)). Either substitutions of single (S286A, S292A, T296A, S328A, and T550A) or of all five PKC-sites (5x-PKC) with alanine suppressed PKC-induced stimulation of ASP(+) uptake, whereas regulation by p56(lck) tyrosine kinase was conserved in all mutants. Remarkably, the apparent affinities for TEA(+), TPA(+), and quinine were changed differently in each mutant (EC(50) in WT, S286A, S292A, T296A, S328A, T550A, and 5x-PKC in mumol: TEA(+): 105, 153, 56, 1135, 484, 498, 518; TPA(+): 0.1, 2.1, 0.3, 1.0, 43, 0.3, 2.2; quinine: 1.5, 3.0, 2.5, 4.8, 81, 7.6, 8.9, respectively). After mutations, no effects of PKC activation on apparent affinity of rOCT1 for these substrates could be detected, in contrast to what was observed in WT. PKC activation had no significant effect on rOCT1 trafficking from intracellular pools to the cell membrane. Substitution of all PKC sites suppressed PKC-induced phosphorylation of rOCT1. In conclusion, it was found that the presence of all five potential PKC phosphorylation sites is necessary for the PKC-induced stimulation of rOCT1. The different effects on the EC(50) values by the different mutations suggest that the large intracellular loop participates in building the substrate binding pocket of rOCT1 or specifically modulates its structure. PMID- 15829704 TI - Albumin activates ERK via EGF receptor in human renal epithelial cells. AB - Emerging clinical and experimental evidence strongly implicates proteinuria in the progression of kidney disease. One pathway involves the activation of NFkappaB by albumin, and it has been demonstrated that the activation of NFkappaB induced by albumin is dependent on mitogen-activated protein kinase ERK1/ERK2. To study the effect of albumin on gene expression, primary human renal tubular cells were exposed in vitro to albumin (1%) for 6 h, and gene expression profiling was performed with the human oligonucleotide microarray, U133A Affymetrix Gene Chip. In all, 223 genes were differentially regulated by albumin, including marked upregulation of the EGF receptor (EGFR) and IL-8. Accordingly, the authors sought to delineate the signaling pathway linking albumin to the EGFR and activation of ERK1/ERK2. It was found that albumin led to a dose- and time-dependent activation of ERK1/ERK2. Treatment with albumin led to EGFR phosphorylation, but the activation of ERK1/ERK2 was prevented by pretreatment of the cells with AG-1478, the EGFR kinase inhibitor, at a dose that inhibited EGF-induced ERK1/ERK2 activation. Exogenously administered reactive oxygen species (ROS) were found to activate ERK1/ERK2 via the EGFR and src tyrosine kinase activity and pretreatment of cells with the antioxidant N-acetylcysteine (NAC) and the NADPH oxidase inhibitor DPI abrogated albumin-induced activation of ERK1/ERK2. The src tyrosine kinase inhibitor, PP2, also inhibited the albumin-induced activation of ERK1/ERK2. Finally, pretreatment with AG-1478, the MEK inhibitor UO126, and NAC prevented the albumin-induced increase in IL-8 expression. The authors conclude that the EGF receptor plays a central role in the signaling pathway that links albumin to the activation of ERK1/ERK2 and increased expression of IL-8. Gene profiling studies suggest that there may be a positive feedback loop through the EGFR that amplifies the response of the proximal tubule cell to albumin. Taken together, these results suggest that the EGFR may be an important treatment target for kidney disease associated with proteinuria. PMID- 15829705 TI - Medicare ESRD prospective payment system: weighing the evidence. PMID- 15829706 TI - Tipping the balance in glomerulonephritis. PMID- 15829707 TI - Chloride transport in the kidney: lessons from human disease and knockout mice. AB - Knockout mouse models and human inherited diseases have provided important new insights into the physiologic role of chloride transport by CLC Cl(-) channels and KCC K-Cl co-transporters. ClC-K/barrtin Cl(-) channels are important for renal salt reabsorption and possibly for acid secretion by intercalated cells. The endosomal ClC-5 protein is crucial for proximal tubular endocytosis. Its disruption in mice and patients with Dent's disease leads to hypercalciuria and kidney stones through a pathologic cascade that may be entirely explained by an impairment of endocytosis. KCC4 is important for recycling Cl(-) for the basolateral anion exchanger in intercalated cells, as is evident from the renal tubular acidosis resulting from its knockout. Finally, both KCC3 and KCC4 are crucial for proximal tubular cell volume regulation. PMID- 15829708 TI - Urinary podocyte loss is a more specific marker of ongoing glomerular damage than proteinuria. AB - Podocyte loss contributes to the development of glomerulosclerosis. Although podocyte detachment has been recognized as a new mechanism of podocyte loss in glomerular diseases, its time course and relationship to disease activity are not known. Urinary excretion of viable podocytes was quantified in two models of transient glomerular injury, i.e., rats with puromycin aminonucleoside-induced nephrosis (PAN) and mesangioproliferative nephropathy (anti-Thy 1.1 nephritis model), as well as in a model of continuous glomerular injury, i.e., hypertensive nephropathy (5/6-nephrectomy model), and in aging rats. The number of glomerular Wilm's tumor (WT)-1-positive podocytes and the glomerular expression of cell cycle proteins in vivo were assessed. Urinary podocyte loss occurred in both primary (PAN) and secondary (anti-Thy 1.1 nephritis) in parallel to the onset of proteinuria. However, subsequently proteinuria persisted despite remission of podocyturia. In continuous glomerular injury, i.e., after 5/6-nephrectomy, podocyturia paralleled the course of proteinuria and of systemic hypertension, whereas no podocyturia became detectable during normal aging (up to 12 mo). Despite podocyte detachment of varying degrees, no decrease in glomerular podocyte counts (i.e., WT-1 positive nuclei) was noted in either disease model. Podocyturia in the PAN and anti-Thy 1.1 nephritis model was preceded by entry of glomerular podocytes into the cell cycle, i.e., cyclin D1, cdc2, and/or proliferating cell nuclear antigen (PCNA) expression. Podocyturia is a widespread phenomenon in glomerular disease and not simply a reflection of proteinuria because it is limited to phases of ongoing glomerular injury. The data suggest that podocyturia may become a more sensitive means to assess the activity of glomerular damage than proteinuria. PMID- 15829709 TI - Renal phenotype of UT-A urea transporter knockout mice. AB - The urea transporters UT-A1 and UT-A3 mediate rapid transepithelial urea transport across the inner medullary collecting duct (IMCD). In a previous study, using a new mouse model in which both UT-A1 and UT-A3 were genetically deleted from the IMCD (UT-A1/3(-/-) mice), we investigated the role of these transporters in the function of the renal inner medulla. Here the authors report a new series of studies investigating more generally the renal phenotype of UT-A1/3(-/-) mice. Pathologic screening of 33 tissues revealed abnormalities in both the testis (increased size) and kidney (decreased size and vascular congestion) of UT-A1/3( /-) mice. Total urinary nitrate and nitrite (NOx) excretion rates in UT-A1/3(-/-) mice were more than double those in wild-type mice. Total renal blood flow was not different between UT-A1/3(-/-) and wild-type mice but underwent a greater percentage decrease in response to NG-Nitro-L-arginine methyl ester hydrochloride (L-NAME) infusion. Whole kidney GFR (FITC-inulin clearance) was not different in UT-A1/3(-/-) mice compared with controls and underwent a similar increase in response to a greater dietary protein intake. Fractional urea excretion was markedly elevated in UT-A1/3(-/-) mice on a 40% protein diet, reaching 102.4 +/- 8.8% of the filtered load, suggesting that there may be active urea secretion somewhere along the renal tubule. Although there was a marked urinary concentrating defect in UT-A1/3(-/-) mice, there was no decrease in aquaporin 2 or aquaporin 3 expression. Furthermore, although urea accumulation in the inner medulla was markedly attenuated, there was no decrease in sodium ion concentration in tissue from outer medulla or two levels of the inner medulla. These results support our conclusion that the urinary concentrating defect in UT A1/3(-/-) mice is caused by a failure of urea transport from the IMCD lumen to the inner medullary interstitium, resulting in osmotic diuresis. PMID- 15829710 TI - Viral infection in the renal transplant recipient. AB - Viruses are among the most common causes of opportunistic infection after transplantation and the most important. The risk for viral infection is a function of the specific virus encountered, the intensity of immune suppression used to prevent graft rejection, and other host factors governing susceptibility. Viral infection, both symptomatic and asymptomatic, causes the "direct effects" of invasive disease and "indirect effects," including immune suppression predisposing to other opportunistic infections and oncogenesis. Rapid and sensitive microbiologic assays for many of the common viruses after transplantation have replaced, for the most part, serologic testing and in vitro cultures for the diagnosis of infection. Furthermore, quantitative molecular tests allow the individualization of antiviral therapies for prevention and treatment of infection. This advance is most prominent in the management of cytomegalovirus, Epstein-Barr, hepatitis B, and hepatitis C viruses. Diagnostic advances have not been accompanied by the development of specific and nontoxic anti-viral agents or effective antiviral vaccines. Vaccines, where available, should be given to patients as early as possible and well in advance of transplantation to optimize the immune response. Studies of viral latency, reactivation, and the cellular effects of viral infection will provide clues for future strategies in prevention and treatment of viral infections. PMID- 15829711 TI - Familial vesicoureteral reflux: testing replication of linkage in seven new multigenerational kindreds. AB - Vesicoureteral reflux (VUR) (OMIM %193000), a common cause of childhood renal failure, is strongly influenced by hereditary factors. Familial VUR most closely conforms to autosomal-dominant inheritance, but because of variable penetrance and expressivity, large multigenerational pedigrees tractable to linkage analysis have been difficult to ascertain. A single genome-wide study of familial VUR has demonstrated linkage to chromosome 1p13, with 78% locus heterogeneity. Previous studies in humans have also suggested loci on chromosomes 6p21, 10q26, and 19q13, whereas mutations in ROBO2 were recently reported in some patients with VUR. Replication of these studies was attempted in seven previously undescribed families from Italy and the United States. Simulation studies, assuming 50% locus heterogeneity, showed that these kindreds had 85% power to replicate linkage and 53% power to achieve genome-wide significance at candidate intervals. Thirty-five markers on chromosomes 1p13, 3p12, 6p21, 10q26, and 19q13 were genotyped and analysis of linkage under a variety of models was performed. Parametric analysis excluded linkage to all candidate loci under genetic homogeneity; moreover, the data did not support statistically significant linkage under models of locus heterogeneity. Similarly, nonparametric, allele-sharing analysis did not reveal any evidence of linkage at any of the loci tested. Thus, despite sufficient power, linkage of familial VUR to previously reported candidate intervals could not be replicated. These data demonstrate substantial genetic heterogeneity of VUR and suggest that mapping strategies relying on a large number of kindreds or single "loaded" pedigrees will be most effective to achieve replication or detection of linkage. PMID- 15829712 TI - Impaired autofeedback regulation of hypothalamic norepinephrine release in experimental uremia. AB - Chronic renal failure (CRF) is associated with multiple hypothalamic dysfunctions, including reduced secretion of gonadotropin-releasing hormone (GnRH). Because GnRH release is tightly controlled by sympathetic neuronal input, a possible alteration of local noradrenergic neurotransmission in experimental CRF was evaluated. Basal, stimulated, and autoinhibited norepinephrine (NE) release was assessed in hypothalamic and hippocampal tissue slices obtained from 5/6-nephrectomized and control rats. Autoinhibition-free NE release from brain slices, prelabeled with [3H]NE and superfused with physiologic buffer, was stimulated by six electrical pulses, 100 Hz (pseudo-one-pulse stimulation). Autoinhibited NE release was induced by 90 pulses at 3 Hz. The release of tritiated NE was measured upon addition of increasing concentrations of unlabeled NE to exogenously activate the inhibitory alpha2-autoreceptor. Although neither basal nor stimulated NE release differed between the groups, significantly lower pIC50 and Imax estimates of the concentration-response curves of exogenous NE on [3H]NE release were observed in CRF rats, suggesting a diminished autoinhibition of hypothalamic noradrenergic terminals in CRF. Western blotting of tissue homogenates disclosed a significantly reduced abundance of alpha2-autoreceptor protein in hypothalamic tissue from CRF rats. These abnormalities were selectively observed in the hypothalamus, whereas noradrenergic autoinhibition seemed unaltered in the hippocampus. The results suggest a diminished autoinhibition of hypothalamic NE release in CRF. Although impaired hypothalamic NE autoinhibition does not explain reduced GnRH secretion in CRF, it may be involved in the pathogenesis of sympathetic hyperactivity associated with this condition. PMID- 15829713 TI - Three-dimensional reconstruction of glomeruli by electron microscopy reveals a distinct restrictive urinary subpodocyte space. AB - For more than 150 years, the only urinary space that has been recognized in the glomerulus conducting primary filtrate to the proximal convoluted tubule has been Bowman's space (BS) (1). Here it is shown that ultrastructural reconstructions of the podocyte and the glomerulus reveal BS to be formed from three distinct urinary spaces through which filtrate must pass before reaching the proximal convoluted tubule. The most restricted region, the subpodocyte space (SPS; first described by Gautier in 1950), was found to cover 58 to 65% of the glomerular filtration barrier. It is morphologically distinct from the rest of BS and has a highly significant restriction to flow based on morphometric measurements. This SPS was altered during increased renal perfusion pressure, consistent with the podocyte dynamically reacting to the increase in filtration. A second anastomosing branching region draining the glomerular center, which has been termed the interpodocyte space, has fewer restrictions to flow into the final region--the shell-like peripheral urinary space. The physiologic role of the restrictive SPS is yet to be determined but likely possibilities include regulation of glomerular filtration and cleaning of the glomerular filtration barrier. PMID- 15829714 TI - Hematopoietic stem cell mobilization therapy accelerates recovery of renal function independent of stem cell contribution. AB - Acute renal failure and tubular cell loss as a result of ischemia constitute major challenges in renal pathophysiology. Increasing evidence suggests important roles for bone marrow stem cells in the regeneration of renal tissue after injury. This study investigated whether the enhanced availability of hematopoietic stem cells, induced by stem cell factor and granulocyte colony stimulating factor, to the injured kidney provides an adequate strategy for stem cell-based therapy to counteract renal ischemia/reperfusion injury. It is interesting that cytokine treatment before injury resulted in significant enhancement of function recovery of the kidney. This, however, was not due to increased incorporation of tubular epithelial cells from bone marrow origin. Importantly, cytokine treatment resulted in impaired influx of granulocytes into the injured kidney. Although cytokine treatment improved renal function rapidly after ischemic injury, the results show that the underlying mechanism likely is not based on stem cell transdifferentiation but rather on altered inflammatory kinetics. PMID- 15829715 TI - The endogenous CXXC motif governs the cadmium sensitivity of the renal Na+/glucose co-transporter. AB - Cadmium (Cd2+) poisoning causes severe renal disorders manifested by defects in reabsorptive transport of various compounds. It is reported here that the renal brush-border membrane Na+/glucose co-transporter-1 (SGLT1) is a molecular target for Cd2+ toxicity. In micromolar concentrations, Cd2+ acted as a noncompetitive, partial inhibitor of methyl-D-glucopyranoside uptake in vesicles from COS-7 cells transiently expressing SGLT1. In contrast, only a modest effect in the closely related Na+/myo-inositol co-transporter-1 (SMIT1) was observed. The factor responsible for this difference was the CXXC motif (X can be any residue) at the cytoplasmic end of the eighth transmembrane segment (TM8) of SGLT1. Thus, a mutational transfer of this motif conveyed Cd2+ sensitivity to SMIT1. Moreover, mimicking the inhibitory effect of Cd2+, the biarsenical molecule FlAsH-EDT2 strongly inhibited the SGLT1 that had an engineered tetracysteine motif at the cytoplasmic end of TM8. The experiments also showed that covalent binding of the sulfhydryl reactive biotin-PEO-maleimide to the SGLT1 wild type but not to the mutant lacking the CXXC motif was suppressed by Cd2+. Taken together, these results suggest that in SGLT1, Cd2+ binding to the CXXC motif induces conformational changes that cause a partial inhibition of d-glucose transport. PMID- 15829716 TI - Health by association. PMID- 15829718 TI - G-CSF, but not corticosterone, mediates circulating neutrophilia induced by febrile-range hyperthermia. AB - We previously showed that sustained exposure to febrile-range hyperthermia (FRH) for 24 h caused an increase in circulating granulocyte colony-stimulating factor (G-CSF) levels and a peripheral neutrophilia in mice (Hasday J, Garrison A, Singh I, Standiford T, Ellis G, Rao S, He JR, Rice P, Frank M, Goldblum S, and Viscardi R. Am J Pathol 162: 2005-2017, 2003). In this study, we utilized a conscious temperature-clamped mouse model to analyze the kinetics of G-CSF expression and peripheral neutrophil expansion and the contributions of FRH-induced G-CSF expression, glucocorticoid generation, and catecholamine-induced neutrophil demargination. In conscious mice housed at an ambient temperature of 34.5 degrees C, core temperature rapidly equilibrated at 39.5-40 degrees C. Peripheral neutrophil counts increased 2-fold after 24-h exposure to hyperthermia, peaked at 3.6-fold baseline levels after 36-h exposure to FRH, and returned to baseline levels after 42 h of sustained hyperthermia. Plasma G-CSF levels were increased by 6.8-fold after 24 h and peaked at 40-fold baseline levels after 36 h in the hyperthermic mice. Plasma corticosterone levels peaked at 3.3-fold baseline levels after 30-h sustained hyperthermia and returned to baseline by 42 h. Immunoneutralization of G-CSF blocked FRH-induced peripheral neutrophilia, but blockade of the glucocorticoid receptor with mifepristone failed to modify FRH induced neutrophilia. Epinephrine induced similar increases in peripheral blood absolute neutrophil counts in euthermic mice (2.2-fold increase) and mice exposed to FRH for 36 h (1.8-fold increase). Collectively, these data suggest that FRH induced expression of G-CSF drives the sustained peripheral neutrophilia that occurs during sustained (36 h) hyperthermia, whereas glucocorticoid generation and catecholamine-induced demargination play little role in this response. PMID- 15829719 TI - Passive mechanics of canine internal abdominal muscles. AB - The internal abdominal muscles are biaxially loaded in vivo, and therefore length tension relations along and transverse to the directions of the muscle fibers are important in understanding their mechanical properties. We hypothesized that 1) internal oblique and transversus abdominis form an internal abdominal composite muscle with altered compliance than that of either muscle individually, and 2) anisotropy, different compliances in orthogonal directions, of internal abdominal composite muscle is less pronounced than that of its individual muscles. To test these hypotheses, in vitro mechanical testing was performed on 5 x 5 cm squares of transversus abdominis, internal oblique, and the two muscles together as a composite. These tissues were harvested from the left lateral side of abdominal muscles of eleven mongrel dogs (15-23 kg) and placed in a bath of oxygenated Krebs solution. Each tissue strip was attached to a biaxial mechanical testing device. Each muscle was passively lengthened and shortened along muscle fibers, transverse to fibers, or simultaneously along and transverse to muscle fibers. Both transversus abdominis and internal oblique muscles demonstrated less extensibility in the direction transverse to muscle fibers than along fibers. Biaxial loading caused a stiffening effect that was greater in the direction along the fibers than transverse to the fibers. Furthermore, the abdominal muscle composite was less compliant than either muscle alone in the direction of the muscle fibers. Taken together, our data suggested that the internal abdominal composite tissue has complex mechanical properties that are dependent on the mechanical properties of internal oblique and transversus abdominis muscles. PMID- 15829720 TI - Transduction of phosphorylated heat shock-related protein 20, HSP20, prevents vasospasm of human umbilical artery smooth muscle. AB - Activation of cyclic nucleotide-dependent signaling pathways inhibits agonist induced contraction of most vascular smooth muscles except human umbilical artery smooth muscle (HUASM). This impaired vasorelaxation may contribute to complications associated with preeclampsia, intrauterine growth restriction, and preterm delivery. Cyclic nucleotide-dependent signaling pathways converge at the phosphorylation of the small heat shock-related protein HSP20, causing relaxation of vascular smooth muscle. We produced recombinant proteins containing a protein transduction domain linked to HSP20 (rTAT-HSP20). Pretreatment of HUASM with in vitro phosphorylated rTAT-HSP20 (rTAT-pHSP20) significantly inhibited serotonin induced contraction, without a decrease in myosin light chain phosphorylation. rTAT-pHSP20 remained phosphorylated upon transduction into isolated HUASM as demonstrated by two-dimensional gel electrophoresis. Transduction of peptide analogs of phospho-HSP20 containing the phosphorylation site on HSP20 and phosphatase-resistant mimics of the phosphorylation site (S16E) also inhibited HUASM contraction. These data suggest that impaired relaxation of HUASM may result from decreased levels of phosphorylated HSP20. Protein transduction can be used to restore intracellular expression levels and the associated physiological response. Transduction of posttranslationally modified substrate proteins represents a proteomic-based therapeutic approach that may be particularly useful when the expression of downstream substrate proteins is downregulated. PMID- 15829722 TI - Biomechanics of the lung parenchyma: critical roles of collagen and mechanical forces. AB - The biomechanical properties of connective tissues play fundamental roles in how mechanical interactions of the body with its environment produce physical forces at the cellular level. It is now recognized that mechanical interactions between cells and the extracellular matrix (ECM) have major regulatory effects on cellular physiology and cell-cycle kinetics that can lead to the reorganization and remodeling of the ECM. The connective tissues are composed of cells and the ECM, which includes water and a variety of biological macromolecules. The macromolecules that are most important in determining the mechanical properties of these tissues are collagen, elastin, and proteoglycans. Among these macromolecules, the most abundant and perhaps most critical for structural integrity is collagen. In this review, we examine how mechanical forces affect the physiological functioning of the lung parenchyma, with special emphasis on the role of collagen. First, we overview the composition of the connective tissue of the lung and their complex structural organization. We then describe how mechanical properties of the parenchyma arise from its composition as well as from the architectural organization of the connective tissue. We argue that, because collagen is the most important load-bearing component of the parenchymal connective tissue, it is also critical in determining the homeostasis and cellular responses to injury. Finally, we overview the interactions between the parenchymal collagen network and cellular remodeling and speculate how mechanotransduction might contribute to disease propagation and the development of small- and large-scale heterogeneities with implications to impaired lung function in emphysema. PMID- 15829723 TI - Mechanical signal transduction in skeletal muscle growth and adaptation. AB - The adaptability of skeletal muscle to changes in the mechanical environment has been well characterized at the tissue and system levels, but the mechanisms through which mechanical signals are transduced to chemical signals that influence muscle growth and metabolism remain largely unidentified. However, several findings have suggested that mechanical signal transduction in muscle may occur through signaling pathways that are shared with insulin-like growth factor (IGF)-I. The involvement of IGF-I-mediated signaling for mechanical signal transduction in muscle was originally suggested by the observations that muscle releases IGF-I on mechanical stimulation, that IGF-I is a potent agent for promoting muscle growth and affecting phenotype, and that IGF-I can function as an autocrine hormone in muscle. Accumulating evidence shows that at least two signaling pathways downstream of IGF-I binding can influence muscle growth and adaptation. Signaling via the calcineurin/nuclear factor of activated T-cell pathway has been shown to have a powerful influence on promoting the slow/type I phenotype in muscle but can also increase muscle mass. Neural stimulation of muscle can activate this pathway, although whether neural activation of the pathway can occur independent of mechanical activation or independent of IGF-I mediated signaling remains to be explored. Signaling via the Akt/mammalian target of rapamycin pathway can also increase muscle growth, and recent findings show that activation of this pathway can occur as a response to mechanical stimulation applied directly to muscle cells, independent of signals derived from other cells. In addition, mechanical activation of mammalian target of rapamycin, Akt, and other downstream signals is apparently independent of autocrine factors, which suggests that activation of the mechanical pathway occurs independent of muscle-mediated IGF-I release. PMID- 15829724 TI - Barometric whole body plethysmography in mice. PMID- 15829725 TI - Amplified fragment length polymorphism-based genetic relationships among weedy Amaranthus species. AB - Weedy Amaranthus species frequently cause economically significant reductions in crop yields. Accurate identification of Amaranthus species is important for efficient weed control, but Amaranthus species can interbreed, which might cause difficulty when identifying hybrid-derived specimens. To determine which of several economically important weedy Amaranthus species are most genetically similar, and thus most likely to produce viable hybrids, we performed amplified fragment length polymorphism (AFLP)-based unweighted pair group method with arithmetic mean (UPGMA) analysis on 8 of these species, with 141 specimens representing 98 accessions. The analysis grouped the specimens into four principal clusters composed of Palmer amaranth (Amaranthus palmeri S. Wats.) and spiny amaranth (Amaranthus spinosus L.); Powell amaranth (Amaranthus powellii S. Wats.), redroot pigweed (Amaranthus retroflexus L.), and smooth pigweed (Amaranthus hybridus L.); waterhemp (Amaranthus tuberculatus (Moq.) Sauer) and sandhills amaranth (Amaranthus arenicola I.M. Johnst.); and tumble pigweed (Amaranthus albus L.). The cluster analysis provided evidence suggesting hybridization among Powell amaranth, redroot pigweed, and smooth pigweed. Further investigations using molecular analysis of the ribosomal internal transcribed spacer region from atypical plants supported this notion. Three species, Palmer amaranth, sandhills amaranth, and waterhemp, are dioecious; nevertheless, the Palmer amaranth and waterhemp-sandhills amaranth clusters were distinct from each other. The Palmer amaranth-spiny amaranth cluster included a cluster of Palmer amaranth and two clusters of spiny amaranth, a monoecious species. Thus the dioecious species Palmer amaranth and waterhemp may not necessarily hybridize with each other more readily than they would to one or more of the monoecious Amaranthus species. PMID- 15829726 TI - Single-locus gametophytic incompatibility in autotetraploids. AB - It is known that a single-locus gametophytic self-incompatibility (GSI) system can persist with just two distinct alleles in an autotetraploid population, in contrast to diploid GSI systems, assuming "competitive interaction" in which heteroallelic pollen is universally compatible. The steady-state population structure of a GSI system in autotetraploids was investigated in an undivided population assuming "competitive interaction." A deterministic model was developed to predict the frequencies of genotypes with two, three, or four distinct S alleles, assuming no mutation or population subdivision. The model showed that unlike in diploid GSI systems, the limiting values of the frequencies of genotype classes do not minimize pollen wastage. PMID- 15829727 TI - The transfer and history of "reduced height genes" (Rht) in wheat from Japan to Europe. AB - Wheat is the main crop and often a strategic crop in many European countries. From a historical perspective, we describe the transfer of "reduced height genes" (Rht genes) from Japanese wheat varieties to wheat varieties in Europe and their influence on the increase of the total wheat production in the last century. Historic pathways of Rht genes were influenced directly or indirectly by wheat breeders exchanging seed samples and by some governments importing large quantities of wheat during historically critical periods for their countries. PMID- 15829728 TI - Peditree: pedigree database analysis and visualization for breeding and science. AB - At the Wageningen Laboratory of Plant Breeding, a software package has been developed to query a simple structured database with variety pedigree data. The package, called Peditree, creates a tree-shaped representation of pedigree information and has several visualization and lookup options. Estimates of inbreeding coefficient within a pedigree or coefficients of coancestry among pedigrees can be obtained. Furthermore trait data--if available--can be linked, displayed within the pedigree tree, and used to highlight pedigree entries that comply with set criteria. PMID- 15829729 TI - Sparse and wavy hair: a new model for hypoplasia of hair follicle and mammary glands on rat chromosome 17. AB - Mutant animals in the skin and hair have been used to identify important genes in biomedical research. We describe a new mutant rat, sparse and wavy hair (swh), that spontaneously arose in a colony of inbred WTC rats. The mutant phenotype was characterized by sparse and wavy hair, which was most prominent at age 3-4 weeks, and was inherited in an autosomal recessive manner. The swh/swh rats showed impaired gain of body weight, and their hair follicles were reduced both in number and size, associated with hypoplasia of the sebaceous glands and the subcutaneous fat tissue. Female swh/swh rats were unable to suckle their offspring. Their mammary glands were hypoplastic, and differentiation of mammary epithelial and myoepithelial cells was impaired. Linkage analysis of 579 backcross rats localized the swh locus to a .35-cM region between D17Rat131 and D17Rat50 in the distal end of rat Chr 17. The swh locus spanned the 3.7-Mb genomic region where 24 genes have been mapped and corresponded to the centromere region of the mouse Chr 2 or the region of the human Chr 10p11.1-p14. None of the genes or loci described in mouse or human hair and skin diseases mapped to these regions. These findings suggest that the rat swh is a novel mutation associated with impaired development of the skin appendages, such as hair follicles, sebaceous glands, and mammary glands, and will provide an experimental model to clarify a gene and mechanisms for development of skin appendages. PMID- 15829730 TI - Mapping quantitative trait loci associated with leaf and stem pubescence in cotton. AB - Leaf pubescence in cotton have a potential for insect pest management. Varying degrees of leaf trichome density in Gossypium species and cultivars have been associated to a series of five genes, referred to as t(1)-t(5). We used two segregating interspecific G. hirsutum x G. barbadense backcross populations developed in our laboratory to assess qualitatively and quantitatively leaf and stem pubescence. QTL analyses were performed using simple and composite interval mapping. Based on both types of measurements and under both types of QTL analyses, nine QTLs met permutation-based thresholds. The nine QTLs mapped to four different chromosome regions. Highest LOD values corresponded to the QTLs detected on c6 (four colocalized QTLs) and on D03 (two QTLs) for which the higher pubescence in the progeny derived from the pubescent G. hirsutum parent alleles. Conversely, on c17 (one QTL) and A01 (two QTLs), the G. hirsutum parental alleles affected negatively pubescence. These results combined with another published study confirm (1) the location in a center region of chromosome 6 of the t(1) locus as a major locus/gene determining leaf pubescence, and (2) additional genes located on seven additional chromosomes have been shown to impart trichome density either positively or negatively. The existence of a high density of PCR based loci in most of the regions identified as harboring leaf pubescence QTLs, particularly that on chromosome 6, will facilitate future efforts for map-based cloning. PMID- 15829731 TI - Time course of top-down and bottom-up influences on syllable processing in the auditory cortex. AB - In speech perception, extraction of meaning from complex streams of sounds is surprisingly fast and efficient. By tracking the neural time course of syllable processing with magnetoencephalography we show that this continuous construction of meaning-based representations is aided by both top-down (context-based) expectations and bottom-up (acoustic-phonetic) cues in the speech signal. Syllables elicited a sustained response at 200-600 ms (N400m) which became most similar to that evoked by words when the expectation for meaningful speech was increased by presenting the syllables among words and sentences or using sentence initial syllables. This word-like cortical processing of meaningless syllables emerged at the build-up of the N400m response, 200-300 ms after speech onset, during the transition from perceptual to lexical-semantic analysis. These findings show that the efficiency of meaning-based analysis of speech is subserved by a cortical system finely tuned to lexically relevant acoustic phonetic and contextual cues. PMID- 15829732 TI - Functional significance of olfactory-induced oscillations in the human amygdala. AB - We recorded directly from the amygdalar nucleus of nine epileptic patients performing a delayed odor-matching recognition memory task. Time-frequency analysis of the responses to the odorants revealed that the stimulations elicited induced oscillatory responses, as well as already described olfactory evoked potentials. These oscillatory responses were composed of two frequency components -one in the beta band (15-25 Hz) and a faster one, in the low gamma band (25-35 Hz)--both of which lasted during the full duration of the inspiration. In pairs of identical odorants, the power of gamma oscillations was weaker for the second odorant (the target) than for the first one (the sample). We observed no such difference when the first and second odorants of a pair were different. Thus, gamma oscillations in the amygdala are weaker for repeated stimuli, a mechanism known as repetition suppression. This is consistent with an involvement of the human amygdala in the encoding and retrieval of olfactory information independently of its hedonic properties, at least in epileptic patients. Altogether, our results corroborate in humans evidence found in animals that oscillations serve as a common coding process of olfactory information. PMID- 15829733 TI - Double dissociation between serotonergic and dopaminergic modulation of medial prefrontal and orbitofrontal cortex during a test of impulsive choice. AB - Dysregulation of the prefrontal cortex (PFC) has been implicated in impulse control disorders, including attention deficit hyperactivity disorder. A growing body of evidence suggests that impulsivity is non-unitary in nature, and recent data indicate that the ventral and dorsal regions of the PFC are differentially involved in distinct aspects of impulsive behaviour, findings which may reflect differences in the monoaminergic regulation of these regions. In the current experiment, levels of dopamine, serotonin and their metabolites were measured in the medial PFC (n = 12) and orbitofrontal cortex (OFC) (n = 19) of rats using in vivo microdialysis during the delay-discounting model of impulsive choice, where impulsivity is defined as selection of small immediate over larger delayed rewards. Yoked groups were also dialysed to control for instrumental responding and reward delivery. Significant increases in 5-hydroxytryptamine efflux were observed in the mPFC, but not in the OFC, during task performance but not under yoked control conditions. In the OFC, 3,4-di-hydroxy-phenylocetic acid (DOPAC) levels increased in animals performing the task but not in yoked animals, whereas mPFC DOPAC levels increased in all subjects. These data suggest a double dissociation between serotonergic and dopaminergic modulation of impulsive decision-making within distinct areas of frontal cortex. PMID- 15829734 TI - Different functional loops between cerebral cortex and the subthalmic area in Parkinson's disease. AB - We investigate the extent to which functional circuits coupling cortical and subthalamic activity are multiple and segregated by frequency in untreated Parkinson's disease (PD). To this end, we recorded EEG and local field potentials (LFPs) from macroelectrodes inserted into the subthalamic nucleus area (SA) in nine awake patients following functional neurosurgery for PD. Patients were studied after overnight withdrawal of medication. Coherence between EEG and SA LFPs was apparent in the theta (3-7 Hz), alpha (8-13 Hz), lower beta (14-20 Hz) and upper beta (21-32 Hz) bands, although activity in the alpha and upper beta bands dominated. Theta coherence predominantly involved mesial and lateral areas, alpha and lower beta coherence the mesial and ipsilateral motor areas, and upper beta coherence the midline cortex. SA LFPs led EEG in the theta band. In contrast, EEG led the depth LFP in the lower and upper beta bands. SA LFP activity in the alpha band could either lead or lag EEG. Thus there are several functional sub-loops between the subthalamic area and cerebral cortical motor regions, distinguished by their frequency, cortical topography and temporal relationships. Tuning to distinct frequencies may provide a means of marking and segregating related processing, over and above any anatomical segregation of processing streams. PMID- 15829736 TI - Inflaming chronic pain. PMID- 15829735 TI - Age-dependent spontaneous hyperexcitability and impairment of GABAergic function in the hippocampus of mice lacking trkB. AB - Patterned intrinsic network activity plays a central role in shaping immature neuronal networks into functional circuits. However, the long-lasting signals that regulate spontaneous activity of developing circuits have not been identified. Here we study the net impact of TrkB signaling on early network activity of identified neuronal populations by analyzing postnatal hippocampi from trkB null mice. Ca2+ imaging showed that pyramidal neurons of trkB-/- mice displayed a decrease in intrinsic synchronous activity in neonatal animals but an increase in juveniles. Strikingly, alterations in network activity in trkB-/- hippocampus were associated with an aberrant induction of the transcription factor Fos. In contrast to pyramidal neurons, spontaneous [Ca2+]i oscillations in trkB-/- interneurons were consistently impaired throughout postnatal development. Moreover, the number of GABAergic synapses and the expression levels of GAD65 and KCC2 were decreased in mutant hippocampi, indicating that pre- and post-synaptic GABAergic components were impaired in trkB-/- mice. Finally, the partial blockade of GABA(A) receptor in postnatal slices revealed that mutant hippocampi displayed an increased susceptibility to network hyperexcitability. These results indicate that the lack of TrkB signaling during development impairs GABAergic neurotransmission, thereby leading to an age-dependent decrease followed by an increase in the intrinsic excitability of neuronal circuits. Furthermore, the present study indicates that long-lasting TrkB signaling may contribute to the construction of CNS circuits by modulating patterns of spontaneous [Ca2+]i oscillations. PMID- 15829737 TI - Time and aging--mechanisms and meanings. AB - A recent European Molecular Biology Laboratory Conference on Science and Society entitled "Time & Aging--Mechanisms & Meanings" fascinated scientists from different research areas as well as nonscientists. Topics discussed included not only the biological aging process but also the psychological effects of aging and social influences that affect this process. PMID- 15829738 TI - Using Controls for Molecular Cytogenetic Testing in Clinical Practice. PMID- 15829739 TI - Use Reference Bands to Accurately Estimate ISCN Band Levels 400, 550, and 850. AB - In their 2002 Guidelines for chromosome analysis of peripheral blood, the American College of Medical Genetics states that "The 550-band stage should be the goal of all constitutional studies..." The College of American Pathologists requires that the average case be analyzed at the 400-band level of resolution for routine work, and that the 550-band level be achieved in appropriate blood samples. The challenge is how to identify the 400, 550, and 850-band levels confidently and consistently. In this study, our objectives were to develop simple and reliable criteria to estimate band level, and to evaluate our laboratory's performance with respect to those criteria. Using the ISCN(1995) ideogram as a reference, candidate bands were selected for the three band levels: 400, 550 and 850. A pilot and two follow-up studies were conducted and a set of candidate bands were validated against the Vancouver method of evaluating band level so that band level scores were similar using either method. The final set of reference bands were the presence of 9q32 and 20q13.2 for the 400-band level; 5q33.2 and 10q22.2 for the 550-band level; and 3p26.1, 18q22.3 and 20q13.32 for the 850-band level. Cell selection improved after each technologist was provided a composite image of chromosomes with reference bands highlighted. The band level criteria presented here involve no band counting, appear to be objective, can help to improve quality and consistency among technologists, and can ensure compliance with regulatory agencies. PMID- 15829740 TI - Changing times, evolving responses. PMID- 15829741 TI - Relationship between clinical presentation and visual outcome in postoperative and posttraumatic endophthalmitis in south central India. AB - PURPOSE: To determine risk factors for poor visual outcome in postoperative and posttraumatic endophthalmitis in a large referral center in south central India. METHODS: In this prospective observational series the authors examined 388 patients of postoperative (n= 206) and posttraumatic (n= 182) endophthalmitis at the L V Prasad Eye Institute in Hyderabad, India between 1991 and 1997. The analysis was confined to 236 patients-128 (62.1%) postoperative and 108 (59.3%) posttraumatic patients who were followed for a minimum period of 3 months. A detailed protocol was followed. Chi-square and logistic regression analysis were used to determine risk factors for visual outcome worse than 6/18 and worse than 6/120. RESULTS: Postoperative endophthalmitis: In univariate analysis the features associated with poor visual acuity (grouped as < 6/18 and < 6/120) included intracapsular cataract surgery, poor presenting visual acuity, presence of vitreous cells, inability to visualise the optic disc on indirect ophthalmoscopy, presence of vitreous membranes on ultrasonography, and a culture positive vitreous biopsy. In the multivariate analysis, visual acuity of less or equal light perception (LP) at presentation was associated with a 3-month postoperative visual acuity of < 6/18, with an odds ratio of 5.85 [1.25 - 27.42, 95% CI], and vitreous membranes seen on ultrasonography was associated with a final visual acuity of < 6/120, with an odds ratio of 2.47 [1.05 - 5.83, 95% CI]. Posttraumatic endophthalmitis: In univariate analysis the features associated with poor visual acuity (grouped as < 6/18 and < 6/120) included a retained intraocular foreign body (IOFB), trauma by needle (hypodermic or sewing), poor presenting visual acuity, inability to visualise the optic disc on indirect ophthalmoscopy, presence of vitreous membranes on ultrasonography, and a culture positive vitreous biopsy. In multivariate analysis, IOFB was associated with a 3 month follow-up visual acuity of < 6/18, with an odds ratio of 5.90 [1.85 - 18.78, 95% CI], and trauma by a needle (hypodermic or sewing) and retained IOFB was associated with a final visual acuity of < 6/120, with an odds ratio of 4.47 [1.22 - 16.38, 95%CI] and 3.76 [1.36 - 10.37, 95% CI] respectively. CONCLUSION: This is the largest, single-centre, prospective study on risk factors for poor visual outcome in postoperative and posttraumatic endophthalmitis. The independent risk factor for 3-month follow-up visual acuity of < 6/18 was the presenting visual acuity of < or =LP in postoperative endophthalmitis and a retained IOFB in posttraumatic endophthalmitis. The independent risk factor for 3 month visual acuity of < 6/120 was the presence of vitreous membranes on ultrasonography in postoperative endophthalmitis, and trauma by a needle (hypodermic/ sewing) and retained IOFB in posttraumatic endophthalmitis. PMID- 15829742 TI - Epidemiological and microbiological diagnosis of suppurative keratitis in Gangetic West Bengal, eastern India. AB - PURPOSE: To determine the epidemiological pattern and risk factors involved in suppurative corneal ulceration in Gangetic West Bengal, eastern India, and to identify the specific microbial agents responsible for corneal infections. METHODS: All patients with suspected microbial keratitis presenting to the corneal clinic at Disha Eye Hospital, Barrackpore, West Bengal, India, from January 2001 to December 2003 were evaluated. Sociodemographic data and information pertaining to the risk factors were recorded. After diagnosing infective corneal ulcer clinically, corneal scraping and cultures were performed. RESULTS: Over a three-year period, 1198 patients with suppurative keratitis were evaluated. Ocular trauma was the most common predisposing factor in 994 (82.9%) patients (P< 0.0001), followed by use of topical corticosteroids in 231 (19.28%) patients. Cultures were positive in 811 (67.7%) patients. Among these culture positive cases, 509 (62.7%) patients had pure fungal infections (P< 0.001), 184 (22.7%) patients had pure bacterial infections and 114 (14.1%) had mixed fungal with bacterial infections. Acanthamoeba was detected in 4 (0.49%) patients. The most common fungal pathogen was Aspergillus spp representing 373 (59.8%) of all positive fungal cultures (P< 0.0001), followed by Fusarium spp in 132 (21.2%) instances. Most common bacterial isolate was Staphylococcus aureus, representing 127 (42.6%) of all the bacterial culture (P< 0.0001) followed by Pseudomonas spp 63 (21.1%). CONCLUSION: Suppurative keratitis in Gangetic West Bengal, most often occurs after a superficial corneal trauma with vegetative or organic materials. Fungal ulcers are more common than bacterial ulcers. Aspergillus spp and Staphylococcus aureus were the most common fungus and bacteria respectively. These "regional" findings have important public health implications for the treatment and prevention of suppurative corneal ulceration in this region of India. PMID- 15829743 TI - Efficacy and safety of latanoprost for glaucoma treatment: a three-month multicentric study in India. AB - PURPOSE: To evaluate the short-term efficacy and safety of 0.005% topical latanoprost in Indian eyes. DESIGN: Prospective non-randomised open-label multicentric trial. METHODS: One hundred and fifty patients with ocular hypertension (OHT), primary open-angle, pseudoexfoliation or pigmentary glaucoma were enrolled at four centers. Each center contributed at least 20 patients. Following baseline measurements, 0.005% latanoprost was applied topically once daily in the evening for three months. Patients were examined at 2, 6 and 12 weeks. The primary outcome measure was mean intraocular pressure (IOP) reduction. The mean diurnal variation of IOP (difference between highest and lowest IOP) at baseline and at 12-weeks was compared. RESULTS: One hundred and thirty of 150 enrolled patients completed the study. One randomly selected eye of each patient was included for analysis. At three months, latanoprost reduced the mean IOP from 24.9 (+/- 3.16) mmHg at baseline to 16.10 (+/- 2.7) mmHg, a reduction of 35.25%. 83% had a reduction in IOP of > 25%. The IOP reduction was maintained throughout the study period, and was not affected by gender or age of the patient. One eye did not show any response to the drug. Daytime diurnal variation of IOP was reduced from 4.5 to 2.9 mmHg. 20 patients had conjunctival hyperemia. Six patients had side effects requiring withdrawal from the study. CONCLUSIONS: In this short-term multicentric study, latanoprost effectively reduced IOP and stabilised the diurnal curve in Indian eyes. There were no clinically significant ocular or systemic adverse effects. PMID- 15829744 TI - Early results of penetrating keratoplasty following limbal stem cell transplantation. AB - PURPOSE: To describe the early results of penetrating keratoplasty (PKP) in patients who had earlier received limbal transplantation (LT). METHODS: Prospective, non-comparative interventional case series comprising of four patients with limbal stem cell deficiency (LSCD) due to chemical injury (Cases 1, 2, 4) and xeroderma pigmentosum (Case 3). Cadaveric kerato-limbal allografts or living-related conjunctival-limbal allografts were done in four eyes followed by PKP for visual rehabilitation 3-4.5 months later. The following details were noted: demographics, primary aetiology, type of limbal transplant (cadaveric or living-related), immunosuppression, vision and ocular surface stability before and after LT and PKP, surgical complications and outcome of PKP. RESULTS: Three eyes received living-related conjunctival-limbal allotransplantation and one received cadaveric kerato-limbal allograft. Duration of follow up after PKP ranged from 4 to 11 months. Visual acuity improved in the early postoperative period in all patients but reduced in 2 due to endothelial rejection and after trans-scleral cyclophotocoagulation for medically uncontrolled glaucoma. The ocular surface remained stable in all patients. All patients were started on immunosuppression on the first postoperative day. This was continued till the last follow-up visit. Post-PKP complications were punctate epithelial keratopathy, corneal allograft rejection and secondary glaucoma (one patient each). CONCLUSION: Satisfactory visual rehabilitation is possible after PKP following LT without compromising ocular surface stability. However, a prolonged and close follow-up is warranted to avert complications. PMID- 15829745 TI - Bilateral epithelial defects after laser in situ keratomileusis. Clinical features, management and outcome. AB - PURPOSE: To describe the preoperative characteristics, intraoperative details, management, and postoperative in patients with bilateral epithelial defects after laser in situ keratomileusis (LASIK). METHODS: Retrospective non-comparative case series. RESULTS: Six patients with bilateral epithelial defects after LASIK were part of a cohort of 605 patients undergoing bilateral LASIK at our center from December 2001 to April 2003. The mean age of the patients (5M:1F) was 28.5 +/- 7.9 years, and the average pretreatment myopic spherical equivalent (SE) refraction was 7.3 +/- 0.7 D (-4, -12.25D). An epithelial flap was present in 6 eyes and an epithelial defect with a mean diameter of 3 mm (2mm, 6mm) was seen in 6 eyes. In four patients the epithelial disturbance was bilaterally similar. All defects occurred in the inferior cornea and the epithelial flaps had the hinge positioned superiorly. None of the patients had ocular or systemic risk factors that could have resulted in this complication. A bandage contact lens was used in 6 eyes. At last follow-up of 5.5 +/- 9.5 months (0.25, 21 months), unaided visual acuity was 6/9 or better in 10 eyes. Best spectacle-corrected visual acuity (BSCVA) was maintained in 8 eyes, while 4 eyes lost one line of BSCVA. Recurrent corneal erosions were not reported in the follow-up period. CONCLUSIONS: These patients represent a hitherto unrecognised group of individuals who appear to have a subclinical weakness of adhesion of the corneal epithelium to the underlying structures, which is not evident on clinical examination. This results in bilateral epithelial disturbances after LASIK. Appropriate management results in satisfactory clinical outcomes. Other options for treatment of the fellow eye of such patients include the use of a different microkeratome, release of suction during the reverse pass of the Hansatome microkeratome, and photorefractive keratectomy if the refractive error is low. PMID- 15829746 TI - Can ocular torsion be measured using the slitlamp biomicroscope? AB - PURPOSE: To evaluate and compare a stereo-biomicroscopic technique of ocular torsion measurement with fundus photographic technique. METHODS: This prospective masked observational study included 72 eyes of 36 consecutive subjects with normal ocular motility. The disc-foveal angle was measured by two masked investigators using a slitlamp stereo-biomicroscopic and fundus photographic technique. RESULTS: The mean age was 13.7 years (6 - 44 years), and 15 (41.6%) were males. The average torsion using the slitlamp technique was 5.5 +/- 3.3 degrees and 6.1 +/- 4.3 degrees with fundus photographic technique. Clinical agreement between the two techniques for 5 degree and 6 degree torsion was 81% and 90% respectively. The average difference between the two techniques was 3.0 +/- 2.6 degrees . The mean inter-ocular difference was 3.3 +/- 2.7 degrees (SD, 2.77) and 5.5 +/- 4.6 degrees using the slitlamp and fundus photographic technique respectively. The Pearson correlation coefficient (r2) was 0.5. CONCLUSION: The fundus photographic technique remains the gold standard test for the measurement of ocular torsion. The slitlamp biomicroscopic method can be useful where a fundus camera is unavailable. Further studies are required to test the repeatability of these tests. PMID- 15829747 TI - Results of probing for congenital nasolacrimal duct obstruction in children older than 13 months of age. AB - PURPOSE: To determine the success rate of probing for congenital nasolacrimal duct obstruction in children older than 13 months of age. METHODS: In a retrospective study of 84 children, 13 months and older who underwent probing for congenital nasolacrimal duct obstruction, the age at probing was correlated with success of probing. The children were divided into two groups, Group 1(13 - 24 months) and Group 2 (>24 months). Success was defined as complete resolution of signs and symptoms. The chi-square test was used to analyse the result. RESULTS: The success rate was 88.1% in-group 1 and 80.1% in-group 2 (P = 0.84). There was no significant difference in the cure rate with increasing age (P = 0.6). CONCLUSION: Probing is highly successful in the older age group and should remain the first line of treatment in older children. PMID- 15829748 TI - Split-conjunctival grafts for double-head pterygium. AB - PURPOSE: To describe the technique of split-conjunctival grafts (SCG) for double head pterygia and to evaluate its postoperative outcome. METHOD: A retrospective analysis of seven eyes with primary double-head pterygium was done. All eyes received pterygium excision with split-conjunctival grafts harvested from the superior quadrant. All patients were followed up for recurrence of the lesion and incidence of complications. RESULTS: No recurrence was noted with a mean follow up of 17.7 +/- 6 months. CONCLUSION: Split-conjunctival graft is a useful procedure for double-head pterygium. PMID- 15829749 TI - Disseminated metastasis following periampullary cancer resection masquerading as uveitis. AB - The Uveitis Masquerade Syndromes (UMS) are a group of ocular diseases that mimic intraocular inflammation, but are in fact neoplastic in nature. We report a patient with disseminated malignancy who presented with uveitis 5 years after an apparently successful resection of periampullary adenocarcinoma. The Masquerade Syndrome was detected by cytological examination of the vitreous. PMID- 15829750 TI - Speciation of coagulase negative staphylococcus causing bacterial keratitis. AB - Thirty-five coagulase negative Staphylococcus (CoNS) cultured from corneal ulcer were speciated and antibiotic sensitivity tested. S epidermidis was the commonest isolate and it was sensitive to ampicillin and vancomycin. PMID- 15829751 TI - RPE atrophy following photodynamic therapy in type 2A idiopathic parafoveal telangiectasis. AB - We report a case of Type 2A idiopathic parafoveal telangiectasis treated by Photodynamic Therapy (PDT), resulting in retinal pigment epithelial (RPE) atrophy corresponding to the size of the laser spot used. This raises a doubt regarding the safety of photodynamic therapy in CNVM secondary to type 2A parafoveal telangiectasis. PMID- 15829752 TI - A case of periocular dirofilariasis masquerading as a lid tumour. AB - We describe an unusual case of periocular Dirofilariasis masquerading as a slow growing lid tumour in an elderly female patient. PMID- 15829753 TI - Persistent depot of triamcinolone acetonide after a single intravitreal injection. AB - The persistence of depot of triamcinolone at four months following a single intravitreal injection is described. PMID- 15829754 TI - A cluster of cases of Mycobacterium chelonei keratitis following penetrating keratoplasty. AB - Cluster infection with Mycobacterium chelonei following penetrating keratoplasty is described. All the donor eyes were harvested from the same collection centre. This underscores the importance of adherence to sterile protocols. PMID- 15829755 TI - Perceived barriers to the provision of low vision services among ophthalmologists in India. AB - PURPOSE: To identify and report the perceived barriers to the provision of low vision services among ophthalmologists in India. METHODS: Seventy nine ophthalmologists responded to a structured self-administered questionnaire. Information was collected to understand the level of awareness and barriers/constraints to provision of low vision services. Significant factors associated with each barrier/constraint and perceptions on providing low vision care were investigated. RESULTS: Lack of training/knowledge [65 (82.3%)], lack of awareness [59 (74.7%)] and non-availability of low vision devices [57 (72.2%)] were perceived as the major constraints / barriers to providing low vision care. At least one significant factor was found for each of the above constraints/barriers in providing low vision care. The perception of lack of awareness as being one of the constraints/barriers was significantly higher [OR 3.97 (95% CI, 1.02 - 7.8)] among ophthalmologists from organisations providing low vision services. The perception of lack of motivation as constraint/barrier was significantly higher [OR 3.62 (95% CI, 1.3 - 10.3)] among ophthalmologists from organisations providing low vision services and/or those involved in VISION 2020: The Right to Sight programmes [OR 3.83 (95% CI, 1.4 - 10.4)]. The likelihood of responding that low vision care is time consuming was greater for those belonging to a teaching institute [OR 7.19 (95% CI, 2.0 - 26.1)], those involved in low vision services [OR 5.45 (95% CI, 1.8 - 16.5)] and those who knew that low vision is a priority in VISION 2020 [OR 15.1, 95% CI, 1.5 -155.4]. CONCLUSION: Ophthalmologists need more education about the benefits of low vision care in order to increase their level of awareness and knowledge. PMID- 15829756 TI - Intravitreal triamcinolone acetonide in serpiginous choroidopathy. PMID- 15829757 TI - Phacolytic glaucoma in Fuch's heterochromic uveitis. PMID- 15829758 TI - Myoconjunctival enucleation for enhanced implant motility. Result of a randomised prospective study. PMID- 15829759 TI - Myoconjunctival enucleation for enhanced implant motility. Result of a randomised prospective study. PMID- 15829760 TI - Guest Editorial: Mental Health and the Workplace: Towards a Research Agenda in Canada. PMID- 15829761 TI - Nature and prevalence of mental illness in the workplace. AB - This discussion paper explores the state of knowledge about the prevalence of mental illness and its effect on the working population. Major trends in the literature are also commented on, and significant gaps in knowledge are identified. Annually, 12% of Canadians from 15 to 64 years suffer from a mental disorder or substance dependence. Few studies have examined the prevalence of mental disorders among Canadian workers. Results from Ontario estimate that monthly, about 8% of the working population has a diagnosable mental disorder. Preliminary findings also indicate differences in the prevalence of mental disorders among workers with regard to occupation, age, sex, physical disorders, work environment and work-related stress. Studies indicate that mental and emotional health problems are associated with staggering social and economic costs, which create a heavy burden on the workplace. About one-third of society's depression-related productivity losses can be attributed to work disruptions. The impact of mental illness on the workplace has been examined in terms of its effect on presenteeism, absenteeism and disability days. The presence of any of these has been used to indicate decreased productivity, the largest burden arising from presenteeism. In total, Canada annually loses about $4.5 billion from this decreased productivity. Mental illness is also associated with short term and long-term disability, which in turn is often related to insurance coverage. Mental illness related disability claims have doubled and mental illness accounts for 30% of disability claims, at a cost of $15 to $33 billion annually. The needs of the working population and employers must be addressed. We must be aware of patterns of mental disorder among occupational groups and industry sectors. In addition, we must understand how the disability benefit structure impacts the prevalence as well as patterns of disability related to mental illness. Effective policies and programs must be based on solid evidence. PMID- 15829762 TI - Essential to understand the relationship between mental illness and work. AB - In their paper "Nature and Prevalence of Mental Illness in the Workplace," Dewa et al. (2004) take on a task of enormous difficulty but of even greater importance. Generally, Dr. Dewa and her colleagues carry out this task extremely well. They define the principal dimensions of the problem clearly and summarize key frontiers of our knowledge. At the same time, as a good discussion paper should, they raise more questions than they answer. I will spend a very brief time commenting on the valuable questions this paper answers - and then devote more time to the questions it raises, questions I believe need to be addressed, and priorities I view as most pressing if we are to advance our understanding of mental illness in the workplace. PMID- 15829763 TI - Workplace prevention and promotion strategies. AB - Psychosocial factors refer to all organizational factors and interpersonal relationships in the workplace that may affect the health of the workers. Currently, two psychosocial risk models are universally recognized for producing solid scientific knowledge regarding the vital link between social or psychological phenomena at work and the development of several diseases, such as cardiovascular diseases or depression. The first is the "job demand-contro support" model, which was defined by Karasek and to which the concept of social support has been added; the second is the "effort/reward imbalance" model defined by Siegrist. The public health perspective calls for theoretical models based on certain psychosocial attributes of the work environment for which there is empirical evidence of their pathogenic potential for exposed workers. Not only do these models reduce the complexity of the psychosocial reality of the work to components that are significant in terms of health risks, but they also facilitate the development and implementation of workplace interventions. Psychosocial risk intervention strategies currently implemented by companies are predominantly individual-oriented and aim chiefly at reducing the effects of stressful work situations by improving individual ability to adapt to the situation and manage stress. Like personal protection equipment for exposure to physical or chemical risks, these secondary prevention measures are commendable but insufficient, because they aim to reduce only the symptoms and not the cause of problems. Any intervention program for these risks should necessarily include a primary prevention component with a view to eliminating, or at least reducing, the psychosocial pathogenic agents in the workplace. Several authors have suggested that well-structured organizational approaches are most effective and should generate more important, longer-lasting effects than individual approaches. However, the evidence should be strengthened by more systematic studies to assess the models, their implementation and the outcomes for employers and employees alike. The research agenda on mental health and the workplace should have the following goals; to foster the development and evaluation of well adapted models of interventions designed to reduce adverse psychosocial factors and their mental health effects to give a better understanding of the prevalence of work organization risk factors in Canada, how they may be changing and how they affect mental health in the long term to acquire an understanding of the effects on mental health of prominent trends in organizational practices, such as restructuring, lean production and flexible staffing (all of which result in precarious employment), that may pose special risks for women, immigrants or aging workers in Canada to collect data on the considerable direct and indirect costs to business, workers and society of work-related stress in Canada. PMID- 15829764 TI - Research must look at what interventions work as well as when and why. AB - This discussion paper is well-written and up to date. A strong feature is its attention to the biological and behavioural mechanisms that connect exposure to certain job characteristics with illness. First, some remarks are made with respect to the definition and measurement of psychosocial risk factors. It is concluded that we do know which factors in work are major risk factors for stress and its consequences for ill health, and that in order to prevent or reduce occupational stress we must first make a reliable and valid diagnosis, identifying these risk factors and risk groups. Next, some issues are discussed regarding future priorities (interventions and monitoring). It is argued that in order to develop further a "psychosocial intervention science," we will need to address three types of questions. PMID- 15829765 TI - Industry recognizes the importance of taking action. AB - Any gap between the demands of the environment and the individual's expectations and resources places his or her health at risk. The individual's perception and evaluation determine the extent of the risk and the degree of negative or positive impact. Companies now recognize the importance of taking action to deal with workplace organization (i.e., primary prevention), in addition to offering more conventional methods of secondary and tertiary prevention. Variables associated with the intervention context, which were identified in Vezina's text, are conditions for the successful implementation of any occupational health and safety project. As representatives of a joint occupational health and safety association, we feel that we should be focusing mainly on primary prevention in the workplace. Research should be conducted to offer companies the procedures, methods, tools and means they need in order to implement effective and sustainable prevention strategies that will maintain a healthy workplace. PMID- 15829766 TI - Organizational and environmental factors can influence effectiveness of new care models. AB - This paper puts forward important concepts related to the diagnosis and treatment of individuals with mental illness and the workplace. These include the need to take an integrated bio-psycho-social approach, an emphasis on functional outcomes, the need to improve the current fragmentation of occupational mental health services, the benefits of identifying individuals at risk of developing mental health problems, the difficulties in disseminating educational information, and the impact of the relationship between workplace and family factors. PMID- 15829767 TI - Workplace culture and mental health are interwoven. AB - Workplace health is a serious issue. Illness and injury take a significant toll on employers, employees and the bottom line. Research has focused primarily on the physical health and injury side of the workplace health equation, with much less attention being paid to the mental health factors that affect employers, employees, families, productivity, efficiency, quality, creativity and innovation. PMID- 15829768 TI - Disability management, return to work and treatment. AB - Individuals with severely disabling mental illness receive more benefit from supported employment initiatives than from other vocational services, but these initiatives show variable job tenure and low implementation by governments. For those with less severely disabling mental illnesses, such as depression, evidence based treatment results in substantial restoration of job function, and restored work function occurs in synchrony with reduced symptomatology. However, there is a substantial degree of residual impairment despite receiving standard treatment. Major research trends include an increasing focus on occupational recovery in less severe forms of mental illness and potential application of integrated disability management models to occupational recovery from disabling mental disorders. Promising research directions include effectiveness of standard mental healthcare in restoring work function; effectiveness of actively managing co morbid mental health problems for disabling physical disorders; population factors affecting return to work in those with disabling mental disorders; identification of policies fostering occupational recovery for disabling mental disorders; effectiveness of innovative mental healthcare focused on occupational recovery; and organizational interventions to foster occupational recovery in employees with disabling mental disorders. PMID- 15829769 TI - Research priorities are critical. AB - Building on the review by Goldner et al., I argue that the first research tasks for disability management in Canada should be to articulate conceptual models; define promising disability management interventions and characterize the current longitudinal health and labour market trajectories of Canadians disabled by mental conditions. Research should next emphasize pilot studies of novel interventions and observational studies that examine the relationship between treatment, workplace interventions, work environments and disability outcomes. Selective randomized trials of disability management interventions should be considered downstream. Mental health treatment optimization should be assigned a lower funding priority than research on workplace interventions. PMID- 15829770 TI - For the employer productivity is critical. AB - The Stakeholder's Response to Disability Management, Return to Work and Treatment highlights the key challenges for researchers from a business perspective: 1. The importance of mental health and disability as business issues. Given this, the research needs to consider the employee population, their challenges and needs. 2. Mental health disability is a multi-faceted concern that requires a research agenda that recognizes interconnections between symptom management, coping skills, individual decision making and opportunity. 3. The goal of disability management is to support ability and provide workers with the opportunity to function productively at work. A "rea-world" focus is essential for research and the resolution of systemic barriers. The challenge is to ensure that knowledge is applicable and considers the sources of motivation and "de-motivation" for both the employees and employers. PMID- 15829771 TI - Stigma and work. AB - This paper addresses what is known about workplace stigma and employment inequity for people with mental and emotional problems. For people with serious mental disorders, studies show profound consequences of stigma, including diminished employability, lack of career advancement and poor quality of working life. People with serious mental illnesses are more likely to be unemployed or to be under-employed in inferior positions that are incommensurate with their skills or training. If they return to work following an illness, they often face hostility and reduced responsibilities. The result may be self-stigma and increased disability. Little is yet known about how workplace stigma affects those with less disabling psychological or emotional problems, even though these are likely to be more prevalent in workplace settings. Despite the heavy burden posed by poor mental health in the workplace, there is no regular source of population data relating to workplace stigma, and no evidence base to support the development of best-practice solutions for workplace anti-stigma programs. Suggestions for research are made in light of these gaps. PMID- 15829772 TI - Best practices in workplace mental health:: an area for expanded research. AB - Mental health, mental illness and the workplace is a timely topic. Many Canadian employers are recognizing the consequences for their bottom line of not addressing the issue. Dr. Stuart's paper was a thorough discussion of the topic of stigma. I suggested four points for consideration. First, the time may have come to substitute the word discrimination for stigma. It opens a rights and responsibilities dialogue that would be valuable. Also, employers and employees understand the term. Second, there are two populations to consider; those who want to enter the workforce, possibly for the first time, and those who want to stay in the workforce. Studying both populations' needs and experiences would yield new knowledge. Third, consider broadening the investigation scope to include instances where things are working (best practices). And finally, concentration only on anti-stigma programs would exclude other innovations. PMID- 15829773 TI - Important to investigate the dynamics of the stigma process. AB - Studies have shown that the stigma of the most common mental disorder, namely depression, expose people with these disorders to a substantial amount of stigmatization in the workplace. Apart from the descriptive assessment of the magnitude of stigma, it is also important to investigate the dynamics of the stigma process. Agreeing with Dr. Stuart, three approaches to research on stigma and the workplace are proposed. The first is the dimension of social stigma, i.e., knowledge, attitudes and practices of employers. The second is the perspectives of the patients, i.e., self- stigmatization. The third is legal and policy frameworks, i.e., structural discrimination. PMID- 15829774 TI - What does it take to transform mental health knowledge into workplace practice? Towards a theory of action. AB - The purpose of this discussion paper is to consider the question of a knowledge base that might undergird a systematic approach to transforming mental health knowledge into workplace practice. Drawing on a range of systems-change and related research, the paper begins by contrasting the nature of "workplace" as opposed to "mental health knowledge" systems. On the basis of the Luhmann's concepts, these systems are cast as fundamentally being determined by the types of communication in which they are engaged (business about business matters, and mental health about mental health matters). For information from one to be adopted by the other requires the translation of mental health concepts into language understood by the workplace, by people capable of understanding both. The paper then examines the importance of determining a vision of desired outcome from such knowledge transfer. What is the desired outcome? A workplace free of mental health problems? The role of both values and existing knowledge in determining these is outlined, along with the importance of engaging the most immediately involved actors in developing the vision. Because valid and reliable knowledge is central to the task, a framework is set out in which the most immediately relevant research findings might be considered in relation to each other. Three categories of knowledge are proposed: employer-led preventive measures, employee-focused workplace interventions and community-based resources supportive of workplaces. Knowledge drawn from summative analyses of existing research is matched against these three categories to illustrate the potential for guiding both implementation and research decision making.The final section draws together the key elements of an active approach to promoting and supporting knowledge transformation from mental health research to workplaces. PMID- 15829775 TI - Bibliometric analysis of research on mental health in the workplace in Canada,1991-2002. AB - This paper uses the Medline biomedical papers database to measure scientific production on mental health in the workplace (MHWP) during the 1991"2002 period at the world, Canadian, provincial, urban, institutional and researcher levels. The level of scientific output has doubled at the world level and tripled at the Canadian level during the last 12 years. At the provincial level, Ontario, Quebec, British Columbia and Alberta are leading in absolute number of papers. Ontario largely dominates both in terms of output and on a per capita basis. At the level of cities, Toronto and Montreal are the largest producers of papers on MHWP. The most important institutions in terms of papers on MHWP are McMaster University, Universite de Montreal, the University of Toronto, the University of British Columbia and the University of Western Ontario. The universities with the largest number of active researchers in MHWP are McMaster University, Universite Laval and York University. PMID- 15829776 TI - Mental health and mental illness in the workplace: diagnostic and treatment issues. AB - Mental health, mental illness and stress-related disability are especially ill defined, complex and controversial issues when considered in the context of the workplace. A multi-determined disorder such as major depressive disorder (MDD) does not fit a simple cause and effect model, but is similar to other complex occupational illnesses such as low back pain. Currently, a knowledge gap exists between mental health professionals and employers regarding symptom-based models of illness and function-based models of work performance. As a result, psychiatric disorders affecting workers are under-identified and under-treated and likely result in unmitigated impairment and disability. The authors examine several conceptual models for workplace mental illness across medical and psychological disciplines and propose a unifying construct. The utility of the existing screening methods for common workplace illnesses and their potential application are reviewed. The challenges of diagnosis and effective treatment of workplace mental illness are highlighted within an "occupational mental health system" with suggestions for future research directions. PMID- 15829777 TI - Effect of the self-monitoring approach on exercise maintenance during cardiac rehabilitation: a randomized, controlled trial. AB - OBJECTIVE: To evaluate the effect of the self-monitoring approach (SMA) on self efficacy for physical activity (SEPA), exercise maintenance, and objective physical activity level over a 6-mo period after a supervised 6-mo cardiac rehabilitation (CR) program. DESIGN: We conducted a randomized, controlled trial with 45 myocardial infarction patients (38 men, seven women; mean age, 64.2 yrs) recruited after completion of an acute-phase, exercise-based CR program. Patients were randomly assigned to an SMA group (n = 24) or control group (n = 21). Along with CR, the subjects in the SMA group self-monitored their weight and physical activity for 6 mos. The SMA used in this study was based on Bandura's self efficacy theory and was designed to enhance confidence for exercise maintenance. The control group participated in CR only. All patients were evaluated with the SEPA assessment tool. Exercise maintenance, SEPA scores, and objective physical activity (average steps per week) as a caloric expenditure were assessed at baseline and during a 6-mo period after the supervised CR program. RESULTS: Mean period from myocardial infarction onset did not differ significantly between the SMA and control groups (12.1 +/- 1.3 vs. 12.2 +/- 1.2 mos, P = 0.692). All patients maintained their exercise routine in the SMA group. Mean SEPA score (90.5 vs. 72.7 points, P < 0.001) and mean objective physical activity (10,458.7 vs. 6922.5 steps/wk, P < 0.001) at 12 mos after myocardial infarction onset were significantly higher in the SMA than control group. SEPA showed significant positive correlation with objective physical activity (r = 0.642, P < 0.001). CONCLUSIONS: SMA during supervised CR may effectively increase exercise maintenance, SEPA, and objective physical activity at 12 mos after myocardial infarction onset. PMID- 15829778 TI - Low-dose interleukin-1 partially counteracts osteogenic protein-1-induced proteoglycan synthesis by adult bovine intervertebral disk cells. AB - OBJECTIVE: Low back pain associated with degenerative disk disease is a common clinical problem that has enormous socioeconomic impact in today's aging population. As an alternative to the surgical removal of the diseased intervertebral disk, the direct application of a purified growth factor into the intervertebral disk may provide physiatrists a valuable tool to halt or slow down disk degeneration. Our goal here is to determine if low levels of interleukin-1 (IL-1), a proinflammatory cytokine present in the degenerating disk, could interfere with the potentially beneficial effects of growth factors on proteoglycan synthesis. New knowledge gained from this study will prove useful in the development of new treatment modalities that take advantage of the stimulatory effects of growth factors such as osteogenic protein-1 (OP-1). DESIGN: This was an in vitro study of proteoglycan accumulation and synthesis by cells from the nucleus pulposus, inner annulus fibrosus, and outer annulus fibrosus in the bovine intervertebral disk. RESULTS: In cells cultured with serum and no additional exogenous growth factor, treatment with low-dose IL-1 does not result in a significant decrease in proteoglycan synthesis. However, in the case of cells stimulated with OP-1, treatment with IL-1 resulted in a statistically significant decrease in sulfur-35-proteoglycan synthesis by cells derived from all three zones of the bovine intervertebral disk (nucleus pulposus, 60.3% [P < 0.0001]; inner annulus fibrosus, 18.4% [P = 0.0330]; outer annulus fibrosus, 12.3% [P = 0.0255]). Proteoglycan accumulation over the 12-day culture period also decreased significantly (nucleus pulposus, 26.8% [P < 0.0001]; inner annulus fibrosus, 15.8% [P = 0.0276]; outer annulus fibrosus, 16.8% [P = 0.0102]). It is worth noting that cells cultured in the presence of both OP-1 and IL-1 synthesized proteoglycan at a faster rate than cells cultured in the presence of IL-1 alone (nucleus pulposus, 58.5% [P < 0.0001]; inner annulus fibrosus, 39.7% [P = 0.0055]; outer annulus fibrosus, 45.1% [P = 0.0164]). Likewise, cells treated with OP-1 and IL-1 accumulated more proteoglycan in their newly formed matrix than cells cultured in the presence of IL-1 alone (nucleus pulposus, 65.3% [P < 0.0001]; inner annulus fibrosus, 60.5% [P = 0.0034]; outer annulus fibrosus, 19.4% (P = 0.0840]). CONCLUSIONS: Intervertebral disk cells that are stimulated by the growth factor OP-1, to increase the rate at which they produce a proteoglycan-rich matrix become more susceptible to the inhibitory effects of the proinflammatory cytokine IL-1 on the rate of proteoglycan synthesis and accumulation in the matrix over time. This notwithstanding, IL-1 at the low dose used did not totally obliterate the stimulatory effects of OP-1 on matrix formation. Consequently, this growth factor may remain partially effective in stimulating disk repair in vivo even when proinflammatory cytokines such as IL-1 are present. PMID- 15829779 TI - Speed-dependent deviations from a straight-ahead path during forward locomotion in healthy individuals. AB - OBJECTIVE: In subjects with unilateral vestibular malfunction, running is associated with less departure from the straight-ahead path than walking. The purpose of this study was to broaden the scope of these observations by investigating whether they can be generalized to healthy subjects. DESIGN: Healthy blindfolded subjects were asked to traverse a 10-m straight path while walking at a self-selected slow speed, walking at a fast speed, or running. Gait speed and mediolateral feet placement were monitored, and departure from the straight-ahead path (path integration) was determined. RESULTS: In healthy subjects, similar to subjects receiving unbalanced vestibular information, departures from the straight-ahead trajectory (namely, path integration) were larger in slow walking than in running. CONCLUSIONS: In healthy subjects, variation in foot placement between the acts of walking and running seem to account for the enhanced path integration found during running. Perhaps consideration of these findings is also relevant for the interpretation of speed dependent path integration in subjects receiving abnormal unilateral vestibular inputs. PMID- 15829780 TI - Effect of plantar micromechanical stimulation on cardiovascular responses to immobility. AB - OBJECTIVE: We investigated the cardiovascular responses of adult women to the influence of extended quiet sitting and the extent to which these responses may be reversed by micromechanical stimulation of the plantar surface. DESIGN: The cardiovascular responses of 20 healthy adult women (mean age, 55.9 +/- 4.45 yrs) were observed during quiet sitting with and without exposure to a plantar-based micromechanical stimulation. Beat-to-beat heart rate via electrocardiogram was acquired along with preexposure and postexposure blood pressures, from which heart rate variability and mean arterial pressure were determined. Seven stimulus frequencies (0, 15, 22, 44, 60, 90, and 120 Hz, all at 0.2 x g, peak to peak) were tested on each subject. RESULTS: Over one-half of the women tested (11/20) exhibited a significant resting tachycardia (mean, 8.3 +/- 0.5 beats/min) with a corresponding decline in their systolic blood pressure (9.45 +/- 1.8 mm Hg) after 20 mins of quiet sitting. Plantar stimulation at 44 Hz (25 mum, peak to peak) was able to completely reverse the effect of immobility in this group, resulting in a heart rate decline of 2.5 beats/min (P < 0.0001) and a decrease of only 1 mm Hg in systolic pressure (P = 0.006). CONCLUSION: We interpret these results to suggest that the immobility of quiet sitting has a profound effect on the cardiovascular systems in a large fraction of otherwise healthy women, perhaps due to inadequate muscle tone leading to venous insufficiency. Simple external stimulation of the plantar surface seems to be capable of preventing these cardiovascular stress-based responses. PMID- 15829781 TI - Effect of trunk flexion on the occupant neck response to anterolateral whiplash impacts. AB - OBJECTIVE: The purpose of this study was to determine the response of the cervical muscles to increasing low-velocity anterolateral impacts with the volunteer's trunk flexed to the right and left. METHODS: A total of 20 healthy volunteers were subjected to left anterolateral impacts of 4.0, 7.6, 10.7, and 13.4 m/sec and, sequentially, with trunk flexed either left or right. Bilateral electromyograms (EMGs) of the sternocleidomastoids, trapezii, and splenii capitis were recorded. DESIGN: At an acceleration of 13.4 m/sec, with the trunk flexed left, the left trapezius generated 48% of its maximal voluntary contraction EMG, whereas the right trapezius (contralateral to the left anterolateral impact) generated 38% of this variable. All other muscle generated or = 30 mm, respectively (chi-squared test, p = 0.006). CONCLUSION: A high prevalence of peripheral retinal degenerations was found in adult Chinese high myopes. The presence of retinal holes was positively correlated with very high myopia of an axial length of > or = 30 mm. PMID- 15829852 TI - The association of astigmatism and spherical refractive error in a high myopia cohort. AB - PURPOSE: The purposes of this study were to determine whether the degree of myopia influences the presence and degree of total astigmatism, and to assess risk factors of astigmatism in patients with familial nonsyndromic severe myopia. METHODS: We performed a retrospective study of 217 subjects from families with two or more subjects from successive generations with a myopic spherical refractive error of at least -5 D or greater in one eye. Mean myopic spherical equivalent was -10 D and the mean age of myopia onset was 7 years. Refractive error measurements were obtained and the association between the degree of myopia and cylinder power was examined by correlation analysis. RESULTS: The prevalence of astigmatism (1.0 D of cylinder) was 36.1%. With-the-rule astigmatism was most common (55.8%), and the majority of astigmats had between 1.0 and 2.5 D of cylinder (77.6%). Statistically significant associations were found between the presence of astigmatism and risk factors of age and the age of myopia onset. In those patients with astigmatism, however, there was a moderate correlation between the degree of spherical equivalent and cylinder power (r = -0.34, p < 0.0001). Younger age (<16 years) (p = 0.03) was associated with higher cylinder power. CONCLUSIONS: In severely myopic patients, there is a high prevalence of astigmatism that is predominantly with-the-rule. The degree of myopic spherical refractive error is correlated with astigmatism severity but is not a risk factor for the presence of astigmatism. PMID- 15829853 TI - Ametropia and ocular biometry in a U.K. university student population. AB - PURPOSE: The prevalence of myopia is known to vary with age, ethnicity, level of education, and socioeconomic status, with a high prevalence reported in university students and in people from East Asian countries. This study determines the prevalence of ametropia in a mixed ethnicity U.K. university student population and compares associated ocular biometric measures. METHODS: Refractive error and related ocular component data were collected on 373 first year U.K. undergraduate students (mean age = 19.55 years +/- 2.99, range = 17-30 years) at the start of the academic year at Aston University, Birmingham, and the University of Bradford, West Yorkshire. The ethnic variation of the students was as follows: white 38.9%, British Asian 58.2%, Chinese 2.1%, and black 0.8%. Noncycloplegic refractive error was measured with an infrared open-field autorefractor, the Shin-Nippon NVision-K 5001 (Shin Nippon, Ryusyo Industrial Co. Ltd, Osaka, Japan). Myopia was defined as a mean spherical equivalent (MSE) less than or equal to -0.50 D. Hyperopia was defined as an MSE greater than or equal to +0.50 D. Axial length, corneal curvature, and anterior chamber depth were measured using the Zeiss IOLMaster (Carl Zeiss, Jena, GmBH). RESULTS: The analysis was carried out only for white and British Asian groups. The overall distribution of refractive error exhibited leptokurtosis, and prevalence levels were similar for white and British Asian (the predominant ethnic group) students across each ametropic group: myopia (50% vs. 53.4%), hyperopia (18.8% vs. 17.3%), and emmetropia (31.2% vs. 29.3%). There were no significant differences in the distribution of ametropia and biometric components between white and British Asian samples. CONCLUSION: The absence of a significant difference in refractive error and ocular components between white and British Asian students exposed to the same educational system is of interest. However, it is clear that a further study incorporating formal epidemiologic methods of analysis is required to address adequately the recent proposal that juvenile myopia develops principally from "myopiagenic" environments and is relatively independent of ethnicity. PMID- 15829854 TI - Refractive status of indigenous people in the northwestern Amazon region of Brazil. AB - PURPOSE: The purpose of this study was to investigate the refractive status of the illiterate indigenous people of the upper Rio Negro region of the Amazon rain forest in northwestern Brazil. METHODS: From an overall sample of 486 people, 259 indigenous people and 78 Brazilians between 12 and 59 years of age with no compromising optical opacities were refracted with cycloplegic retinoscopy. Subjects were categorized as indigenous if they had at least three generations of indigenous ancestry with no folklore suggesting other ancestors. RESULTS: Myopia was rare among the indigenous population. Only 2.7% of eyes showed myopia of 1.00 D or more and 1.6% (four people) had bilateral myopia of -1.00 D or more. Half of this small group were the only educated indigenous people examined. The prevalence of astigmatism and anisometropia equal to or >1.00 D was 15.5% and 8.2%, respectively. Most of the astigmatism in the indigenous people had an against-the-rule axis. Age was not associated with the refractive errors of the indigenous people. Brazilians from the small city in which the study was performed had higher rates of myopia (6.4% of eyes and 5.1% of subjects bilaterally). Older preeducation adults also had a very low prevalence of myopia (3.2% of eyes and 2.0% of subjects), whereas the younger, slightly educated Brazilians had a higher prevalence of myopia (11.3% of eyes and 9.7% of subjects). CONCLUSION: The low prevalence of myopia in the illiterate indigenous people is consistent with other studies and suggests that myopia is related to literacy. The generational change among the local mixed race Brazilians further supports this conclusion. The relatively high rates of astigmatism and anisometropia in the indigenous people were unusual for a predominantly emmetropic sample. PMID- 15829855 TI - Accommodation, accommodative convergence, and response AC/A ratios before and at the onset of myopia in children. AB - PURPOSE: The purpose of this study was to investigate accommodation, accommodative convergence, and AC/A ratios before and at the onset of myopia in children. METHODS: Refractive error, accommodation, and phorias were measured annually over a period of 3 years in 80 6- to 18-year-old children (mean age at first visit = 11.1 years), including 26 who acquired myopia of at least -0.50 D and 54 who remained emmetropic (-0.25 to + 0.75 D). Refraction was measured by noncycloplegic distance retinoscopy. Concomitant measures of accommodation and phorias were taken for letter targets at 4.0 m and 0.33 m using the Canon R-1 open field-of-view autorefractor with an attached motorized Risley prism and Maddox rod. The accommodation and phoria measurements were used to calculate response AC/A ratios. RESULTS: Compared with children who remained emmetropic, those who became myopic had elevated response AC/A ratios at 1 and 2 years before the onset of myopia, in addition to at onset and 1 year later (t's = -2.97 to 4.04, p < 0.01 at all times). The significantly higher AC/A ratios in the children who became myopic are a result of significantly reduced accommodation. Accommodative convergence was significantly greater in myopes only at onset. CONCLUSIONS: These findings suggest that the abnormal oculomotor factors found before the onset of myopia may contribute to myopigenesis by producing hyperopic retinal defocus when a child is engaged in near-viewing tasks. PMID- 15829856 TI - The effect of monocular and binocular viewing on the accommodation response to real targets in emmetropia and myopia. AB - PURPOSE: Decreased blur-sensitivity found in myopia has been linked with reduced accommodation responses and myopigenesis. Although the mechanism for myopia progression remains unclear, it is commonly known that myopic patients rarely report near visual symptoms and are generally very sensitive to small changes in their distance prescription. This experiment investigated the effect of monocular and binocular viewing on static and dynamic accommodation in emmetropes and myopes for real targets to monitor whether inaccuracies in the myopic accommodation response are maintained when a full set of visual cues, including size and disparity, is available. METHODS: Monocular and binocular steady-state accommodation responses were measured with a Canon R1 autorefractor for target vergences ranging from 0-5 D in emmetropes (EMM), late-onset myopes (LOM), and early-onset myopes (EOM). Dynamic closed-loop accommodation responses for a stationary target at 0.25 m and step stimuli of two different magnitudes were recorded for both monocular and binocular viewing. RESULTS: All refractive groups showed similar accommodation stimulus response curves consistent with previously published data. Viewing a stationary near target monocularly, LOMs demonstrated slightly larger accommodation microfluctuations compared with EMMs and EOMs; however, this difference was absent under binocular viewing conditions. Dynamic accommodation step responses revealed significantly (p < 0.05) longer response times for the myopic subject groups for a number of step stimuli. No significant difference in either reaction time or the number of correct responses for a given number of step-vergence changes was found between the myopic groups and EMMs. CONCLUSION: When viewing real targets with size and disparity cues available, no significant differences in the accuracy of static and dynamic accommodation responses were found among EMM, EOM, and LOM. The results suggest that corrected myopes do not experience dioptric blur levels that are substantially different from emmetropes when they view free space targets. PMID- 15829857 TI - Assessment of objective and subjective eccentric refraction. AB - PURPOSE: When performing perimetry, refracting subjects with central visual field loss, and in emmetropization studies, it is important to accurately measure peripheral refractive errors. Traditional methods for foveal refraction often give uncertain results in eccentric angles as a result of the large aberrations and the reduced retinal function. The aim of this study is therefore to compare and evaluate four methods for eccentric refraction. METHODS: Four eccentric methods were tested on 50 healthy subjects: one novel subjective procedure, optimizing the detection contrast sensitivity with different trial lenses, and three objective ones: photorefraction with a PowerRefractor, wavefront measurements with a Hartmann-Shack sensor, and retinoscopy. The peripheral refractive error in the horizontal nasal visual field of the right eye was measured in 20 degrees and 30 degrees. RESULTS: In general, the eccentric refraction methods compared reasonably well. However, the following differences were noted. Retinoscopy showed a significant difference from the other methods in the axis of astigmatism. In 30 degree eccentric angle, it was not possible to measure 15 of the subjects with the PowerRefractor and the instrument also tended to underestimate high myopia (<-6 D). The Hartmann-Shack sensor showed a myopic shift of approximately 0.5 D in both eccentricities. The subjective method had a relatively larger spread. CONCLUSIONS: This study indicates that it is possible to assess the eccentric refraction with all methods. However, the Hartmann-Shack technique was found to be the most useful method. The agreement between the objective methods and the subjective eccentric refraction shows that detection contrast sensitivity in the periphery is affected by relatively small amounts of defocus. PMID- 15829858 TI - Early ametropia and rod photoreceptor function in retinopathy of prematurity. AB - PURPOSE: Early ametropia, particularly myopia, is frequent in children with a history of preterm birth and retinopathy of prematurity (ROP). The retina is known to govern eye growth and refractive development. We tested the hypothesis that deficits in retinal function are significantly associated with early ametropia in ROP subjects. METHODS: Scotopic electoretinogram (ERG) responses to full field stimuli were studied in 40 ROP subjects aged 8 weeks to 18 years. The ROP was categorized as treated, untreated, or none. Refractive development of each ROP subject was monitored and compared with normal for age. The rod photoresponse parameters were calculated and the postreceptoral responses derived. The ERG parameters in the ROP subjects were compared with normal values for age. RESULTS: Twelve ROP subjects developed early ametropia, 10 myopia, and two hyperopia. In the majority of ROP subjects, receptoral and postreceptoral response parameters were below the normal mean for age. In the 12 children with early ametropia, rod photoreceptor sensitivity was significantly lower than in emmetropic ROP subjects; and in five tested in infancy, deficits in rod photoreceptor sensitivity antedated development of ametropia. The myopic control subjects had no deficits in response parameters. CONCLUSIONS: Retinal dysfunction is significantly associated with early ametropia in these ROP subjects. Thus, mechanisms for the development of ametropia in ROP subjects may involve rod and rod-mediated postreceptoral activity. PMID- 15829859 TI - Temporal integration characteristics of the axial and choroidal responses to myopic defocus induced by prior form deprivation versus positive spectacle lens wear in chickens. AB - PURPOSE: In chicks, the temporal response characteristics to form deprivation and to spectacle lens wear (myopic and hyperopic defocus) show essential differences, suggesting that the emmetropization system "weights" the visual signals differently. To further explore how the eye integrates opposing visual signals, we examined the responses to myopic defocus induced by prior form deprivation vs. that induced by positive spectacle lenses, in both cases alternating with form deprivation. METHODS: Three experimental paradigms were used: 1) Form deprivation was induced by monocular occluders for 7 days. Over the subsequent 7 days, the occluders were removed daily for 12 hours (n = 13), 4 hours (n = 7), 2 hours (n = 7), or 0 hours (n = 6). 2) Birds were form-deprived on day 12. Over the subsequent 7 days, occluders were replaced with a +10 D lens for 2 hours per day (n = 13). 3) Starting at day 11, a +10 D lens was placed over one eye for 2 hours (n = 13), 3 hours (n = 5), or 6 hours (n = 10) per day and were otherwise untreated. Ocular dimensions were measured with high-frequency A-scan ultrasonography; refractive errors were measured by streak retinoscopy at various intervals. RESULTS: In recovering eyes, 2 hours per day of myopic defocus was as effective as 12 hours at inducing refractive and axial recovery (change in refractive error: +10 D vs. +13 D, respectively). By contrast, 2 hours of lens induced defocus (alternating with form deprivation) was not sufficient to induce refractive or axial compensation (change in refractive error: -1.7 D). When myopic defocus alternated with unrestricted vision, 6 hours per day were sufficient to induce nearly full compensation (2 hours vs. 6 hours: 4.4 D vs. 8.2 D; p < 0.0005). Choroids showed rapid increases in thickness to the daily episodes of myopic defocus; these resulted in "long-term" thickness changes in recovering eyes and eyes wearing lenses for 3 or 6 hours per day. CONCLUSIONS: The response to myopic defocus induced by prior form deprivation is more robust than the response induced by positive lenses, suggesting that the underlying mechanisms differ. Presumably, this difference is related to the size of the eye at the onset. Compensatory decreases in growth rate occur without full compensatory choroidal thickening. PMID- 15829860 TI - Adaptability of myopic children to progressive addition lenses with a modified fitting protocol in the Correction of Myopia Evaluation Trial (COMET). AB - PURPOSE: The purpose of this study was to evaluate the adaptability of myopic children participating in the Correction of Myopia Evaluation Trial (COMET) to the use of progressive addition lenses (PALs) with a modified fitting protocol of setting the distance fitting cross 4 mm above the pupil center. METHODS: COMET was a multicenter, randomized clinical trial to evaluate whether PALs vs. single vision lenses (SVLs) slowed the rate of progression in children with juvenile onset myopia over 3 years. Of the 469 children enrolled, 234 were assigned to SVLs and 235 were assigned to PALs (+2.00 D near addition), which were fit according to a standard study protocol with the fitting cross positioned 4 mm above the pupil center. The children's adaptability to the use of PALs was evaluated by comparing the frequency and reasons for problem visits, visual symptoms, cover test results, and adherence to wearing glasses between treatment groups. RESULTS: During the first 3 years of follow-up, no differences were observed between children wearing PALs and those wearing SVLs with respect to the number or reasons for problem visits. One week after receipt of the study glasses, children wearing PALs showed a higher frequency of three visual symptoms related to adaptability: looking down from the blackboard and getting items on their desk in focus (p = 0.001), blur when reading (p = 0.003), and difficulty going down steps (p = 0.02). Children wearing PALs were more likely to report at least one adaptation symptom at 1 week: odds ratio of 2.76 (95% confidence interval = 1.28-5.95). By 1 month, these differences disappeared and the frequency of all visual symptoms remained low and similar for both treatment groups over 3 years. Strabismus was observed on cover test in five children (three SVLs, two PALs). None of the children had to be changed from PALs to SVLs, whereas two of the children were changed from SVLs to PALs as a result of binocular vision problems. The children and parents reported adherence to wearing the eyeglasses approached 100% compliance throughout the follow-up period for both groups. Most (98%) of the 235 children assigned to PALs maintained the modified fitting protocol without any problems over 3 years. CONCLUSION: These data show that children can safely and comfortably wear PALs for at least 3 years. PMID- 15829861 TI - Evaluating the self-esteem of myopic children over a three-year period: The COMET Experience. AB - PURPOSE: The purposes of this study were to evaluate self-esteem over 3 years in the 469 myopic children participating in the Correction of Myopia Evaluation Trial (COMET), and to examine its relationship with lens assignment (progressive addition lenses [PALs] vs. single-vision lenses [SVLs]), myopia progression, and several other ocular and demographic characteristics. METHODS: Data collection included refractive error measurements, child-reported visual symptoms, attitude toward glasses, adherence, and self-esteem as measured by the Self-Perception Profile for Children (SPPC). A two-way analysis of variance (treatment group x time) was performed to examine whether PAL and SVL wearers differed in self esteem over time. Multiple regression analyses were used to evaluate associations between self-esteem at follow-up and relevant factors identified by univariate analyses. RESULTS: Regardless of lens assignment or myopia progression, COMET children reported moderate to high levels of self-esteem at follow-up in the areas of scholastic and athletic competence, physical appearance, social acceptance, behavioral conduct, and global self-worth. Mean scores ranged from 2.87 (+/- 0.68) on athletic competence to 3.40 (+/- 0.56) on global self-worth. Self-esteem changed significantly (p < 0.05) over 3 years in the domains of scholastic competence, social acceptance, and physical appearance. Self-esteem at follow-up was associated with visual symptoms, attitude toward glasses, age, gender, and ethnicity. CONCLUSIONS: Lens assignment and myopia progression were not associated with self-esteem in the COMET cohort. These children had high levels of self-esteem, suggesting that having myopia does not negatively impact self-esteem. Follow-up reports will monitor self-esteem and related factors in this cohort of myopic children over the course of adolescence and early adulthood. PMID- 15829862 TI - National plan for reliable tuberculosis laboratory services using a systems approach. Recommendations from CDC and the Association of Public Health Laboratories Task Force on Tuberculosis Laboratory Services. AB - Since the mid-1990s, public health laboratories have improved tuberculosis (TB) test performance, which has contributed to the resumption of the decline in TB incidence in the United States. However, to eliminate TB in the United States, further improvements are needed in laboratory services to support TB treatment, prevention, and control. A critical step is the development of an integrated system that ensures prompt and reliable laboratory testing and flow of information among laboratorians, clinicians, and TB-control officials. Challenges to developing such a system include 1) establishing lines of communication among laboratorians, clinicians, and TB-control officials; 2) expediting reporting of laboratory results, which can avoid delayed or inappropriate treatment and missed opportunities to prevent transmission; 3) developing evidence-based recommendations for use of new laboratory technologies; 4) maintaining staff proficiency in light of declining numbers of specimens to test, workforce shortages, and loss of laboratory expertise; and 5) upgrading laboratory information systems and connecting all partners. The report of the Association of Public Health Laboratories Task Force presents a framework to improve the future of TB laboratory services and describes the role of the laboratory in TB treatment and control, Task Force processes, general principles and benchmarks, and steps for the dissemination of the Task Force recommendations. This MMWR expands on the Task Force report by describing specific actions and performance measures to guide development and implementation of an integrated system for providing TB laboratory services. CDC and the Association of Public Health Laboratories have developed these guidelines so that laboratorians, clinicians, public health officials, administrators, and funding entities can work together to ensure that health-care providers and TB-control officials have the information needed to treat TB patients, prevent TB transmission, and ultimately eliminate TB in the United States. PMID- 15829863 TI - Health concerns associated with disaster victim identification after a tsunami- Thailand, December 26, 2004-March 31, 2005. AB - The number of persons confirmed dead from the Indian Ocean tsunami that struck on December 26, 2004, had exceeded 174,000 as of March 31, 2005; the majority of decedents were buried or cremated without being identified. In contrast, in Thailand, disaster victim identification (DVI) continues, with approximately 1,800 persons identified among the 5,395 persons confirmed dead; of the dead, approximately 50% were not citizens of Thailand. This large-scale, multinational effort faced immediate challenges, including establishment of four temporary morgues, implementation of safeguards against environmental and occupational health hazards, and coordination of forensic procedures and safety protocols among Thai and international forensic teams. Public health and other agencies performing large-scale DVI in temporary morgues might consider implementing the recommendations and procedures described in this report. PMID- 15829864 TI - Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food--10 sites, United States, 2004. AB - Foodborne illnesses are a substantial health burden in the United States. The Foodborne Diseases Active Surveillance Network (FoodNet) of CDC's Emerging Infections Program collects data from 10 U.S. sites on diseases caused by enteric pathogens transmitted commonly through food. FoodNet quantifies and monitors the incidence of these infections by conducting active, population-based surveillance for laboratory-diagnosed illness. This report describes preliminary surveillance data for 2004 and compares them with baseline data from the period 1996-1998. The 2004 data indicate declines in the incidence of infections caused by Campylobacter, Cryptosporidium, Shiga toxin-producing Escherichia coli (STEC) O157, Listeria, Salmonella, and Yersinia. Declines in Campylobacter and Listeria incidence are approaching national health objectives (objectives 10-1a through 1d); for the first time, the incidence of STEC O157 infections in FoodNet is below the 2010 target. However, further efforts are needed to sustain these declines and to improve prevention of foodborne infections; efforts should be enhanced to reduce pathogens in food animal reservoirs and to prevent contamination of produce. PMID- 15829865 TI - Acute public health consequences of methamphetamine laboratories--16 states, January 2000-June 2004. AB - Methamphetamine (meth), a powerfully addictive stimulant, can be easily produced in illicit, makeshift laboratories and generally is considered the fastest growing illicit drug in the United States. Aside from the inherent physical and physiological dangers of the drug itself, persons in and around meth laboratories can be acutely exposed to hazardous substances used in meth production. Exposure to these substances can occur from volatile air emissions, spills, fires, and explosions. This report describes examples of meth-associated events, summarizes the events reported to the Agency for Toxic Substances and Disease Registry (ATSDR), and suggests injury prevention recommendations, such as how to recognize and properly respond to meth laboratories. PMID- 15829866 TI - Anhydrous ammonia thefts and releases associated with illicit methamphetamine production--16 states, January 2000-June 2004. AB - Anhydrous ammonia, a colorless gas with a pungent, suffocating fumes, is used primarily as an agricultural fertilizer and industrial refrigerant. Anhydrous ammonia is also a key ingredient for illicit methamphetamine (meth) production in makeshift laboratories. Exposure to anhydrous ammonia can be immediately dangerous to life or health. Anhydrous ammonia generally is not available for sale to the public; states require a license for purchase. Because of this, many illicit meth producers (i.e., "cookers") resort to stealing anhydrous ammonia. If released into the environment, anhydrous ammonia can cause acute injuries to emergency responders, the public, and the cookers themselves. In addition, when handled improperly, anhydrous ammonia can be explosive and deadly. This report describes examples of anhydrous ammonia thefts associated with illicit meth production, summarizes ammonia theft events reported to the Agency for Toxic Substances and Disease Registry (ATSDR), and suggests injury prevention recommendations, such as installing valve locks or fencing on unattended tanks and donning appropriate personal protective equipment (PPE) when responding to releases. PMID- 15829867 TI - Peripheral ameloblastoma: a case report. AB - Peripheral ameloblastoma is a rare odontogenic soft tissue tumor, derived from epithelial and/or mesenchymal elements being part of the tooth-forming apparatus. The lesion responses for approximately 1% to 5% of all cases of ameloblastoma affecting alveolar mucosa and gingiva occur, mainly, in the middle age. This article describes a case of peripheral ameloblastoma involving a 20-year-old male located in the (upper/lower, vestibular/buccal) gingiva. After the case presentation, clinical and microscopic findings are discussed. PMID- 15829868 TI - Hypohidrotic ectodermal dysplasia with bilateral impacted teeth at the coronoid process: a case rehabilitated with mini dental implants. AB - Bilateral migration of teeth into the coronoid process in a patient with ectodermal dysplasia has not been reported in the literature except one report in which severe hypodontia and bilaterally ectopic impacted teeth in the coronoid processes of a nonsyndromic patient occurred. This article presents a 15-year-old female with hypohidrotic ectodermal dysplasia who had surgical removal of bilaterally impacted teeth in the coronoid process and was rehabilitated with a dental implant-retained fixed prosthesis in the mandible and over-denture in the maxilla. PMID- 15829869 TI - Raising standards: digital interoperability and DICOM. PMID- 15829870 TI - Possible molecular approach to the treatment of odontogenic keratocyst. PMID- 15829871 TI - Radiolucent lesion of the anterior mandible. PMID- 15829872 TI - Reliability of sentinel lymph node biopsy with squamous cell carcinoma of the oral cavity. AB - OBJECTIVES: Although sentinel lymph node biopsy (SLNB) may reduce surgery-related complications related to unnecessary lymph node dissection and is now widely used for many patients with cutaneous melanoma and breast cancer, its use for oral cancer patients remains controversial. One of the main reasons for the reluctance to initiate SLNB for oral cancer is that the frequency of skip metastasis has not been clarified. The objectives of this study are to examine the frequency of skip metastasis and to evaluate SLNB for oral cancer. STUDY DESIGN: To shed light on these concerns, we first conducted a retrospective study of 296 patients with squamous cell carcinoma of the oral cavity who underwent neck dissection. Next, the accuracy of lymph node biopsy with and without detecting sentinel lymph node was examined. RESULTS: Ten patients showed skip neck metastasis in the level III V region without level I-II involvement. Of these patients, 7 underwent neck dissection when their initially N0 neck progressed to N+, 2 underwent neck dissection when local recurrence occurred, and only 1 underwent surgery as an initial therapy. Most patients who underwent neck dissection as the initial therapy showed skip metastasis. Intraoperative lymph node biopsy without any attempt to detect sentinel lymph nodes by means of blue dye or lymphoscintigraphy was performed on 68 patients with oral cancer. Sixty-one (90%) were diagnosed correctly, whereas 7 diagnosed as N- actually had neck metastasis. SLNB with blue dye was performed on 21 patients. In 17 of them, sentinel lymph node was easily detected, resulting in a correct diagnosis for 16 patients (94%), while 1 with a false negative result actually had micrometastasis. CONCLUSION: These findings seem to suggest that SLNB is useful and can be applied to patients with oral cancer who undergo surgery as the initial therapy. PMID- 15829874 TI - Incidence of cystic changes in radiographically normal impacted lower third molar follicles. AB - OBJECTIVE: The objective of this study was to investigate the cystic changes in radiographically normal follicles associated with impacted lower third molars (ILTM). The relation between the cystic changes and both the angular position and the contact of ILTM with the adjacent tooth was also evaluated. STUDY DESIGN: Follicular spaces of patients who were referred to our clinic for removal of ILTM for a variety of reasons were measured from panoramic radiographs by each author independently. Pericoronal tissues of radiographically normal impacted third molars were examined histopathologically. Then the data were analyzed for associations with age, sex, angular position, and contact of the ILTM with the second molar. RESULTS: There were 30 male and 64 female patients, ages ranging from 14 to 45 years (mean = 21.11). Forty-seven (50%) of the 94 specimens showed cystic changes as the only pathological condition other than inflammation. Most of the cystic changes occurred in the 20-25 year age range. The relation between cystic changes and angular position was statistically significant (chi 2 = 7.46, P < .05). The higher probability of cystic changes was found in the vertically positioned ILTM. The relation between the cystic changes and communication of ILTM with the second molar was not statistically significant (chi 2 = 2.068, P > .05). CONCLUSIONS: Cystic changes may be encountered in the histopathologic examination of asymptomatic third molars, especially in patients older than 20 years. Vertically positioned ILTM especially may be suspected for the higher possibility of cystic changes. PMID- 15829875 TI - Oral piercings among first-year university students. AB - OBJECTIVE: The aim of the study was to examine oral piercings among first-year university students. STUDY DESIGN: First-year university students in 2002 were invited to a dental examination (n = 234; 49 men and 185 women). Students with piercings formed the study group and the rest served as controls. The methods included decayed, missing, and filled teeth (DMF) index, stimulated salivary flow rates, panoramic tomograms, and questionnaires including the Depression Inventory of Beck. Fisher's 2-sided exact test was used for statistical analysis. RESULTS: The prevalence of oral piercings was 3.4%. In the DMF indices, no statistically significant differences existed between the groups. Increased salivary flow rates were noted among students with piercings (63% vs 26%, P < .05). Use of tobacco and illicit drugs, and also depression, were more prevalent in the study group than in the controls. CONCLUSION: Because of the possibility of oral implications, follow-up of oral piercings is essential. PMID- 15829876 TI - Buccal mucosal transposition flap for reconstruction of oropharyngeal-oral cavity defects: an analysis of six cases. AB - BACKGROUND: Operation on tumors situated in the posterior part of the oral cavity is a challenging aspect of head and neck surgery. Both the approach and the postablative reconstruction of the remaining defect require special procedures. An arsenal of reconstructive methods are available depending on the size and complexity of the defect. The goal should be to employ the simplest surgical reconstructive method, resulting in the best functional result and allowing for later complex oncotherapy. OBJECTIVE: To study the outcomes of patients treated using buccal mucosal transposition flaps for reconstruction of posterior oral cavity and oropharyngeal border defects. STUDY DESIGN: A total of 6 patients with defects of the posterior oral cavity and mesopharynx were treated with buccal mucosal transposition flaps. The patients were followed for up to 24 months. RESULTS: Defects ranging from 12 to 27 cm 2 were reconstructed with the buccal mucosal transposition flap. One patient developed an orocutaneous fistula and 1 had some trismus requiring a Z-plasty scar reorientation. One patient suffered from nasal regurgitation. Five of the 6 patients lived past 24 months, the time of follow-up of this study. CONCLUSIONS: The buccal mucosal transposition flap, with its random pattern of circulation, used alone proved to be a safe method to reconstruct soft tissue defects or line exposed hard tissues located in the posterior oral cavity and oropharynx. PMID- 15829877 TI - Metastatic renal cell carcinoma presenting as a parotid tumor. AB - Metastases of malignancies to the parotid region are relatively infrequent (21% 42% of all malignant tumors 1 ), but metastases of infraclavicular origin are infrequent (0.16%-4% 1,2 ). From 1986 to 1998, only 17 cases of parotid metastases of renal clear cell carcinoma were documented in the literature reviewed. In this paper we report a case of a patient with a parotid tumor which was the first manifestation of distant disease. It was only once the tumor was surgically removed that the histopathology allowed the diagnosis of the primary tumor; namely a renal clear cell adenocarcinoma. PMID- 15829878 TI - Prevalence of post-traumatic stress disorder symptoms in orofacial pain patients. AB - OBJECTIVE: There is a high comorbidity between symptoms of post-traumatic stress disorder (PTSD) and chronic pain incidence. The objective of this investigation was to determine the prevalence of PTSD symptoms in chronic orofacial pain patients. STUDY DESIGN: The study included 1478 adult patients (mean age 36.4 +/- 12.7 years) with primary diagnoses of masticatory/cervical muscle pain or temporomandibular joint pain. Patients completed a battery of psychometric questionnaires including a screening for PTSD symptoms. The sample was divided into a PTSD-positive group (n=218, 15%) a PTSD-negative group (n=551, 37%), and a no-stressor group (n=709, 48%) according to stressor incidence and symptom severity. RESULTS: The current prevalence of PTSD symptomatology was considerably higher than that reported in surveys from the general population. Patients in the PTSD-positive symptom group reported significantly higher psychological distress, sleep dysfunction, and pain severity compared to patients in the other groups. Psychological distress as measured by the SCL-90-R reached clinically significant levels only in those patients with PTSD symptomatology. CONCLUSIONS: The results of this study performed at a tertiary care center suggest that TMD patients without PTSD symptomatology show low levels of psychological distress, if any. Clinically significant levels of psychological distress are likely indicators for PTSD. PTSD screening should be included as part of a routine psychometric test battery in TMD patients. PMID- 15829879 TI - Cowden's syndrome affecting the mouth, gastrointestinal, and central nervous system: a case report and review of the literature. AB - Cowden's syndrome (CS) is a rare genodermatosis, of autosomal dominant inheritance and variable phenotype, principally characterized by the formation of hamartomas in various organs, including skin, thyroid, breast, brain, and gastrointestinal tract and by the increased risk for the development of malignancy. The present report details the features of a very rare presentation of Cowden's syndrome affecting not only the mouth and gastrointestinal mucosa but also the central nervous system. PMID- 15829880 TI - Medical considerations in dental treatment of children with Williams syndrome. AB - Williams syndrome is a rare disorder that was first described in 1961. It is thought to be caused by a microdeletion in the long arm of chromosome 7 at 7q11.23 and is a multisystem, congenital, and panethnic disorder characterized by a number of developmental and physical abnormalities, including congenital cardiovascular abnormalities, mental retardation and neurological features, growth deficiency, genitourinary manifestation, gastrointestinal and musculoskeletal problems, behavioral characteristics, craniofacial features, ophthalmologic features, and dental problems. We describe cases of children with Williams syndrome treated in the department of Pediatric Dentistry of the Hadassah School of Dental Medicine, Jerusalem, Israel. The different treatments rendered to these children are discussed followed by general remarks drawn from those treatments and from a literature review. We conclude that sedation can be helpful in the younger age group to reduce anxiety and uncooperative behavior during minimal dental treatments. Treatment under general anesthesia seems more appropriate for older children and adolescents. Special attention should be given to initial evaluation of these patients, especially because with age aortic stenosis tends to intensify, which together with the progressive renal impairment can escalate blood pressure elevation. PMID- 15829882 TI - Salivary heterotopia of the parathyroid gland: a report of two cases and review of the literature. AB - Two cases of periparathyroid salivary gland heterotopia are described. A review of the records of the Department of Pathology, Long Island Jewish Medical Center, over a 4-year period, identified 759 surgical specimens containing parathyroid gland tissue. Of these, 2 (0.26%) contained foci of ectopic salivary gland tissue. Both cases were associated with cyst formation. To date, 9 additional cases of heterotopic salivary gland tissue associated with the parathyroid gland have been described in the literature. PMID- 15829881 TI - Myoepithelioma of the tongue. AB - Soft tissue myoepitheliomas, in contrast to salivary gland myoepitheliomas, are benign neoplasms that typically occur in the soft tissues of the extremities. Both are characterized by a multilobular proliferation of polygonal to fusiform cells embedded in a variably myxoid to chondromyxoid matrix. A histologically similar lesion that has a marked predilection for the anterior dorsum of the tongue has been referred to as ectomesenchymal chondromyxoid tumor. The morphological and immunohistochemical resemblance of soft tissue myoepitheliomas to ectomesenchymal chondromyxoid tumors has led to the use of these designations interchangeably. We present a case of myoepithelioma of the tongue and review the literature, with emphasis on the differential diagnosis and histogenesis of this lesion and pertinent nosologic considerations. PMID- 15829883 TI - Palatal perforation associated with intranasal prescription narcotic abuse. AB - BACKGROUND: Palatal perforation resulting from insufflation of cocaine has been well documented. In comparison, reports of destructive orofacial lesions resulting from intranasal abuse of prescription narcotics are rare. We present the clinical and histologic findings in a case of palatal perforation arising in a patient abusing a prescription opioid drug. The patient denied any history of cocaine use but admitted to habitually crushing and snorting a hydrocodone/acetaminophen preparation. Study design The patient presented to our clinic seeking resolution of speech difficulties associated with an oroantral fistula. Surgical repair of the defect had been attempted unsuccessfully in the past. In addition to blood and chemistry panels, endoscopic examination was conducted, with removal of several biopsy specimens for histologic evaluation and flow cytometry. Biopsy specimens included both lesional and perilesional tissue from within the oral and nasopharyngeal cavities. Culture and cytology for fungal organisms were also performed. RESULTS: Histopathologic examination revealed normal mucosa with diffuse and focal inflammatory changes and no evidence of malignancy. Polarizable foreign material was noted in the specimens. The absence of lymphoid neoplasia was confirmed by flow cytometric analysis. The toxicology panel was positive for the presence of opiates in the blood. Culture and cytology were positive for candidal organisms. A palatal obturator was fabricated for the patient, producing significant improvement in the quality of speech. CONCLUSIONS: This may represent a case of palatal perforation resulting from abuse of a drug other than cocaine. The potential for drugs other than cocaine to produce destructive orofacial lesions should be considered. PMID- 15829884 TI - A 4-year prospective study on epidemiology and clinicopathological presentation of odontogenic tumors in Tanzania. AB - This study, conducted between 1999 and 2003, included all patients with odontogenic tumors referred from all dental clinics and other health facilities in Tanzania to the only 4 referral centers. Demographic data and clinical and histopathologic findings were recorded. Histopathologic specimens were examined by expert pathologists. Ameloblastoma had the highest relative frequency (80.1%) and an incidence rate of 0.68 per million, followed by odontogenic myxoma (7.0%) with an incidence rate of 0.07 per million. Follow-ups of between 6 months and 4 years were carried out. Two patients developed recurrences of ameloblastoma. The incidence of ameloblastoma in this exclusively black African population did not differ from that of European countries. This is contrary to the belief that ameloblastoma has a higher incidence in black Africans as compared to Caucasians. PMID- 15829885 TI - Effect of delayed scanning of storage phosphor plates. AB - OBJECTIVE: To test longevity of image quality in storage phosphor plates (SPPs) at various exposure settings, storage conditions, and delays in scanning. STUDY DESIGN: Fifteen Digora plates were exposed from 0.08 to 0.20 seconds and scanned immediately, 10, 30, and 60 minutes, and 24 hours after exposure. Plates were stored both in daylight and in a light-tight box. Mean gray values (MGVs) were compared using 2 x 5 x 5 factorial ANOVA. Interaction between variables was tested using Bonferroni/Dunn multiple comparisons test. RESULTS: MGVs decreased with increase in exposure but increased with the scan delay. Only MGVs of plates scanned within 10 minutes after exposure were not significantly different from the ones scanned immediately ( P > .05). MGVs increased with scan delay for all exposure times no matter how the plates were stored ( P < .05). CONCLUSION: Based on the time delays examined, it is recommended to scan the Digora SPP no later than 10 minutes after exposure. Longer periods may cause loss of quality. PMID- 15829886 TI - Effects of scintillator on the modulation transfer function (MTF) of a digital imaging system. AB - OBJECTIVE: To investigate the effects of 2 components (scintillator and x-ray generator) in the imaging chain on the modulation transfer function (MTF) of a charge-coupled device (CCD) digital intraoral radiographic system. STUDY DESIGN: Three screens composed of 3 different scintillator materials, namely europium doped lutetium oxide transparent optical ceramic (TOC), thallium-doped cesium iodide (CsI), and terbium-doped gadolinium oxysulfide (GOS), were compared. Each was used, in turn, in conjunction with a CCD detector having a pixel dimension of 19.5 mum. Two different x-ray generators were also used to evaluate this variable. MTF was investigated using the slanted slit method. RESULTS: The TOC provided a good modulation response for low and middle frequencies, reducing to 0 only at a high cutoff frequency. With CsI and GOS, the system MTF dropped to 0 at a lower cutoff frequency than was the case with TOC. Hence, TOC provided higher spatial resolution than the other 2 scintillators tested under the experimental conditions applied. The differences in MTF attributed to the scintillator type were proportional and consistent. CONCLUSIONS: Despite constant pixel dimensions, MTF was affected to a considerable degree by the scintillator applied and the x ray generator used in conjunction with the same CCD imaging device. TOC shows potential as a possible replacement for CsI and GOS as a scintillator screen material for intraoral digital x-ray imaging using a solid-state detector, providing higher spatial resolution under the given experimental conditions. PMID- 15829887 TI - Disc and condyle translation in patients with temporomandibular disorder. AB - OBJECTIVE: The purpose of this study was to elucidate causal relationship between disc and condyle range of movement and clinical signs and symptoms in patients with temporomandibular disorders (TMD), using magnetic resonance imaging (MRI). STUDY DESIGN: The subjects comprised of a study group of 191 patients with TMD and a control group of 43 asymptomatic patients. The clinical assessment consisted of range of maximum mouth opening (MMO) and preauricular pain during mandibular function. After clinical and radiographic findings assessment, disc and condyle condition were examined by MRI and the range of movement was accordingly classified. RESULTS: Disc displacement was observed in 156/191 (81.7%) of the study group and 9/43 (20.9%) of the control group. When disc and condyle mobility was presented around the eminence, wider MMO range was maintained, P < .05. Presence of osteoarthrosis (OA) was not correlated with preauricular pain, because OA variables were mild in the study group. CONCLUSION: Maintenance of disc/condyle translation is an important factor in TMJ function, irrespective of disc displacement or arthritis. PMID- 15829888 TI - A quality assurance program in dental radiographic units in western Greece. AB - OBJECTIVE: This survey reports on successful efforts to establish a quality assurance (QA) program for 50 intraoral x-ray units in the public and private sectors in the region of Achaia, Greece. It was conducted in 2 phases, in 1996 and in 2003, including on-site inspections, QA tests, and standard questionnaires. The aim of the study was to assess equipment conditions, knowledge, and adoption of radiographic QA guidelines by general dentists and, more importantly, the impact of the recommendations and training provided. STUDY DESIGN: The tested parameters focused on radiation protection, equipment maintenance, film speed used, film processing conditions, and radiologic characteristics such as voltage, radiation leakage, type of collimation, source to-skin distance, timer accuracy, and entrance dose. RESULTS: The data gathered in 1996 demonstrate minimal compliance with equipment function requirements and radiation safety measures. 1 The comparative evaluation of all the parameters gathered from the 2 surveys, indicated that in 2003 the vast majority of the dentists followed the recommendations given in 1996. Only 2 dentists persisted in neglecting the guidelines. PMID- 15829889 TI - Effect of rotary or manual instrumentation, with or without a calcium hydroxide/1% chlorhexidine intracanal dressing, on the healing of experimentally induced chronic periapical lesions. AB - OBJECTIVE: To evaluate the healing of experimentally induced chronic periapical lesions in dogs at 30, 75, and 120 days after root canal instrumentation with rotary NiTi files or manual K-files, with or without a calcium hydroxide/1% chlorhexidine paste intracanal dressing. STUDY DESIGN: The second, third, and fourth mandibular premolars and the second and third maxillary premolars of 5 dogs (12 to 18 months of age, weighing 8 to 15 kg) were selected for treatment (a total of 82 root canals). After pulp removal, the root canals were left exposed to the oral cavity for 7 days to allow microbial contamination, after which the root canals were sealed with ZOE cement until periapical lesions were confirmed with radiography. Group I and II teeth were instrumented with manual K-files using the crown-down technique. In group III and IV teeth, NiTi rotary files were used. The apical delta was perforated by using #20 to #30 K-files at the length of the tooth, thus creating a standardized apical opening. The apical stop was enlarged to size 70, with 2.5% sodium hypochlorite irrigation at each file change. Teeth in groups II and IV were dressed with calcium hydroxide (Ca(OH) 2 )/1% chlorhexidine (CHX) paste for 15 days before root filling. Group I and III teeth did not receive an intracanal dressing. The access openings of the teeth were permanently restored with silver amalgam condensed on a glass ionomer cement base. Pairs of standardized periapical radiographs were taken at the beginning of the treatment (0 days) and at 30, 75, and 120 days after filling. RESULTS: There was no significant difference in the rate of radiographic healing of the periapical lesions between manual and rotary instrumentation. Radiographs taken at 120 days showed that the treatment with Ca(OH) 2 /1% CHX paste resulted in a significant reduction in mean size of the periapical lesions in comparison to single-session treatment. These findings were also true for histologic observations. CONCLUSION: The findings support the hypothesis that, regardless of the instrumentation technique (manual or rotary), the use of an intracanal dressing is important in the endodontic treatment of dog's teeth with experimentally induced chronic periapical lesions. PMID- 15829890 TI - Genotoxicity of antimicrobial endodontic compounds by single cell gel (comet) assay in Chinese hamster ovary (CHO) cells. AB - OBJECTIVE: In the current study, the potential DNA damage associated with exposure to a number of antimicrobial endodontic compounds was assessed by the single cell gel (comet) assay in vitro. STUDY DESIGN: Chinese hamster ovary (CHO) cells were exposed to formocresol, paramonochlorophenol, calcium hydroxide, or chlorhexidine at final concentration ranging from 0.01% to 1%. RESULTS: Formocresol, paramonochlorophenol, and calcium hydroxide, as well as chlorhexidine in all concentrations tested did not contribute to the DNA damage. CONCLUSION: These findings are clinically relevant since they represent an important contribution to the correct evaluation of the potential health risk associated with exposure to dental agents. PMID- 15829891 TI - Differences in prevalence of selected bacterial species in primary endodontic infections from two distinct geographic locations. AB - OBJECTIVE: This study intended to compare the prevalence of 7 putative endodontic pathogens in samples of primary endodontic infections taken from patients of 2 distant geographic locations. STUDY DESIGN: Samples from infected root canals associated with asymptomatic periradicular lesions or from pus aspirated from acute periradicular abscesses were collected from patients in Rio de Janeiro, Brazil, and Seoul, South Korea. South Korean samples were frozen and delivered to Brazil, where all steps in the molecular analysis were performed. DNA was extracted and a species-specific nested polymerase chain reaction assay was used to detect 7 target bacterial species. RESULTS: The most prevalent species detected in Brazilian samples were Porphyromonas endodontalis (79% of the cases) , Treponema denticola (79%), and Dialister pneumosintes (76%). The most prevalent species found in South Korean samples were Fusobacterium nucleatum (38% of the cases) , Tannerella forsythia (26%), and Treponema maltophilum (24%). Overall, P endodontalis, D pneumosintes, Filifactor alocis, T denticola, and T forsythia were significantly more detected in Brazilian samples than in South Korean samples ( P < .05). CONCLUSIONS: Findings indicated that the prevalence of some species in infections of endodontic origin may significantly differ from one geographic location to another. PMID- 15829892 TI - Arsenic release provided by MTA and Portland cement. AB - OBJECTIVE: The aim of the present study was to determine the release of arsenic from 2 gray Portland cements, a white Portland cement, and 2 MTAs (ProRoot and MTA-Angelus). STUDY DESIGN: The materials were manipulated and placed in plastic tubes, and the tubes were immersed in glass flasks containing water with grade reagent, pH 5.0. After 3 and 168 h, the water in which the material had been immersed was analyzed regarding the presence of arsenic by atomic absorption spectrophotometry with hydride generation. RESULTS: The levels of arsenic released were similar for Portland cements and MTAs, and were well below those considered to be harmful. CONCLUSIONS: MTA and Portland cements showed very low arsenic presence. The results suggest that MTAs and Portland cements are safe for use in clinical practice in terms of the presence of arsenic. PMID- 15829899 TI - New approaches in the intensive management of cardiovascular risk in the metabolic syndrome. AB - When risk factors such as dyslipidemia and hypertension are inadequately controlled in subjects with the metabolic syndrome by lifestyle interventions, pharmacologic approaches are warranted. Statins are first-line pharmacotherapy for dyslipidemia due to their efficacy for lowering low-density lipoprotein (LDL) cholesterol and may also improve high-density lipoprotein (HDL) cholesterol and triglyceride levels. Fibrates and niacin may be useful in combination with a statin for additionally lowering triglycerides or raising HDL cholesterol. Adequate control of hypertension will usually require two or more drugs; agents that block the renin-angiotensin system are particularly useful in this population, given their demonstrated benefits for reducing the burden of cardiovascular events and end-stage renal disease independent of blood-pressure lowering. A multifaceted approach to risk factor management for the metabolic syndrome will have benefits for prevention of type 2 diabetes and cardiovascular disease. PMID- 15829906 TI - Massive tsunami in Indian Ocean coasts. PMID- 15829907 TI - The National Response Plan: Health and Human Services the lead for Emergency Support Function #8. AB - On January 6, 2005, the National Response Plan (NRP) was introduced to standardize a national approach for responding to natural or man-made threats. The underlying structure for the NRP is the National Incident Management System, which establishes standardized training, organization, and communications procedures that can be used by multiple jurisdictions to interact in a disaster. The NRP clearly identifies authority and leadership responsibilities. The NRP organizes the government's emergency operations into 15 emergency support functions, of which Emergency Support Function #8 pertains to public health and medical services. The United States Public Health Service (PHS) Commissioned Corps is one of the seven uniformed services of the United States, and PHS officers can be deployed to help meet urgent public health needs when traditional mechanisms and resources are overwhelmed. The role of PHS nurses is presented. PMID- 15829908 TI - The threat of communicable diseases following natural disasters: a public health response. AB - Natural disasters, such as the recent Indian Ocean tsunami, can have a rapid onset, broad impact, and produce many factors that work synergistically to increase the risk of morbidity and mortality caused by communicable diseases. The primary goal of emergency health interventions is to prevent epidemics and improve deteriorating health conditions among the population affected. Morbidity and mortality due to infectious diseases can be minimized providing these intervention efforts are implemented in a timely and coordinated fashion. This article presents a review of some of the major issues relevant to preparedness and response for natural disasters. PMID- 15829909 TI - Medical Reserve Corps: strengthening public health and improving preparedness. AB - Across the United States, millions of Americans volunteer their time and efforts to improve the social fabric of their communities. Inevitably, some of these volunteers will be medical and public health professionals. However, because of the complexities of the health field, including concerns about credentialing, training and legal protections, many of these persons have not been able to volunteer in their professional capacities. The terrorist events of 2001 showed that not only would individuals with medical and public health expertise want to volunteer, but that their help could be very much needed in future mass catastrophic events. The Medical Reserve Corps Program was created as a national system of community-based units to promote the local identification, recruitment, training, and activation of volunteers, especially those with medical and public health backgrounds. These Medical Reserve Corps units supplement the existing public health and emergency response entities in the community. PMID- 15829910 TI - The National Disaster Medical System--America's medical readiness force. PMID- 15829912 TI - Statistical prediction of tissue fate in acute ischemic brain injury. AB - An algorithm was developed to statistically predict ischemic tissue fate on a pixel-by-pixel basis. Quantitative high-resolution (200 x 200 microm) cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) were measured on acute stroke rats subjected to permanent middle cerebral artery occlusion and an automated clustering (ISODATA) technique was used to classify ischemic tissue types. Probability and probability density profiles were derived from a training data set (n=6) and probability maps of risk of subsequent infarction were computed in another group of animals (n=6) as ischemia progressed. Predictions were applied to overall tissue fate. Performance measures (sensitivity, specificity, and receiver operating characteristic) showed that prediction made based on combined ADC+CBF data outperformed those based on ADC or CBF data alone. At the optimal operating points, combined ADC+CBF predicted tissue infarction with 86%+/-4% sensitivity and 89%+/-6% specificity. More importantly, probability of infarct (P(I)) for different ISODATA-derived ischemic tissue types were also computed: (1) For the 'normal' cluster in the ischemic right hemisphere, P(I) based on combined ADC+CBF data (P(I)[ADC+CBF]) accurately reflected tissue fate, whereas P(I)[ADC] and P(I)[CBF] overestimated infarct probability. (2) For the 'perfusion-diffusion mismatch' cluster, P(I)[ADC+CBF] accurately predicted tissue fate, whereas P(I)[ADC] underestimated and P(I)[CBF] overestimated infarct probability. (3) For the core cluster, P(I)[ADC+CBF], P(I)[ADC], and P(I)[CBF] prediction were high and similar ( approximately 90%). This study shows an algorithm to statistically predict overall, normal, ischemic core, and 'penumbral' tissue fate using early quantitative perfusion and diffusion information. It is suggested that this approach can be applied to stroke patients in a computationally inexpensive manner. PMID- 15829913 TI - HIV-1 Tat protein-induced alterations of ZO-1 expression are mediated by redox regulated ERK 1/2 activation. AB - HIV-1 Tat protein plays an important role in inducing monocyte infiltration into the brain and may alter the structure and functions of the blood-brain barrier (BBB). The BBB serves as a frontline defense system, protecting the central nervous system from infected monocytes entering the brain. Therefore, the aim of the present study was to examine the mechanisms of Tat effect on the integrity of the BBB in the mouse brain. Tat was injected into the right hippocampi of C57BL/6 mice and expression of tight junction protein zonula occludens-1 (ZO-1) was determined in control and treated mice. Tat administration resulted in decreased mRNA levels of ZO-1 and marked disruption of ZO-1 continuity. These changes were associated with accumulation of inflammatory cells in brain tissue of Tat-treated mice. Further experiments indicated that Tat-mediated alterations of redox related signaling may be responsible for decreased ZO-1 expression. Specifically, injections with Tat resulted in activation of extracellular signal-regulated kinase 1/2 (ERK 1/2) and pretreatment with U 0126, a specific inhibitor of ERK kinase, effectively ameliorated the Tat-induced diminished ZO-1 levels. In addition, administration of N-acetylcysteine (NAC), a precursor of glutathione and a potent antioxidant, attenuated both Tat-induced ERK 1/2 activation and alterations in ZO-1 expression. These results indicate that Tat-induced oxidative stress can play an important role in affecting the integrity of the BBB through the ERK 1/2 pathway. PMID- 15829914 TI - Superoxide dismutase 1 overexpression reduces MCP-1 and MIP-1 alpha expression after transient focal cerebral ischemia. AB - Proinflammatory cytokines and chemokines are quickly upregulated in response to ischemia/reperfusion (I/R) injury; however, the relationship between I/R-induced oxidative stress and cytokine/chemokine expression has not been elucidated. We investigated the temporal profile of cytokine and chemokine gene expression in transient focal cerebral ischemia using complementary DNA array technology. Among 96 genes studied, 10, 4, 11, and 5 genes were increased at 6, 12, 24, and 72 h of reperfusion, respectively, whereas, 4, 11, 8, and 21 genes, respectively, were decreased. To clarify the relationship between chemokines and oxidative stress, we compared the gene and protein expression of monocyte chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein-1 alpha (MIP-1 alpha) in wild-type (WT) mice and copper/zinc-superoxide dismutase (SOD 1) transgenic (Tg) mice. Monocyte chemoattractant protein-1 and MIP-1 alpha mRNA were significantly upregulated at 6 to 12 h of reperfusion. In the SOD 1 Tg mice, however, MCP-1 and MIP-1 alpha mRNA expression was significantly decreased 12 h postinsult. In the WT mice, MCP-1 and MIP-1 alpha protein expression peaked 24 h after onset of reperfusion determined by immunohistochemistry. In the SOD 1 Tg mice, MCP-1 and MIP-1 alpha immunopositive cells were reduced, as were concentrations of these proteins (measured by enzyme-linked immunosorbent assay) at 24 h of reperfusion. Our results suggest that MCP-1 and MIP-1 alpha expression is influenced by I/R induced oxidative stress after transient focal stroke. PMID- 15829915 TI - Oxidative stress transiently decreases the IKK complex (IKKalpha, beta, and gamma), an upstream component of NF-kappaB signaling, after transient focal cerebral ischemia in mice. AB - Nuclear factor-kappaB (NF-kappaB) has a central role in coordinating the expression of a wide variety of genes that control cerebral ischemia. Although there has been intense research on NF-kappaB, its mechanisms in the ischemic brain have not been clearly elucidated. We investigated the temporal profile of NF-kappaB-related genes using a complementary DNA array method in wild-type mice and human copper/zinc-superoxide dismutase transgenic (SOD 1 Tg) mice that had low-level reactive oxygen species (ROS) by scavenging superoxide. Our DNA array showed that IkappaB kinase (IKK) complex (IKKalpha, beta, and gamma) mRNA in the wild-type mice was decreased as early as 1 h after reperfusion, after 30 mins of transient focal cerebral ischemia (tFCI). In contrast, tFCI in the SOD1 Tg mice caused an increase in the IKK complex. The IKK complex protein levels were also drastically decreased at 1 h in the wild-type mice, but did not change in the SOD 1 Tg mice throughout the 7 days. Electrophoretic mobility shift assay revealed activation of NF-kappaB DNA binding after tFCI in the wild-type mice. Nuclear factor-kappaB activation occurred at the same time, as did the phosphorylation and degradation of the inhibitory protein IkappaBalpha. However, SOD 1 prevented NF-kappaB activation, and phosphorylation and degradation of IkappaBalpha after tFCI. Superoxide production and ubiquitinated protein in the SOD 1 Tg mice were also lower than in the wild-type mice after tFCI. These results suggest that ROS are implicated in transient downregulation of IKKalpha, beta, and gamma in cerebral ischemia. PMID- 15829916 TI - Noradrenergic agonists and antagonists influence migration of cortical spreading depression in rat-a possible mechanism of migraine prophylaxis and prevention of postischemic neuronal damage. AB - Cortical spreading depression (CSD) is thought to be a neuronal mechanism that expands the penumbra zone after focal brain ischemia and that causes migraine aura. Both adrenergic agonists and antagonists significantly influence the size of the penumbra zone and decline the frequency of migraine. To study whether these compounds act by influencing CSD, we applied different drugs topically to an area of the exposed cortex of anesthetized adult rats and observed the migration of CSD-related DC potential deflections across the treated area. The adrenergic agonist norepinephrine (1 mmol/L) and the alpha(2)-agonist clonidine (0.56 mmol/L) blocked reversibly the migration of CSD. The beta-blocker propranolol (250 micromol/L to 1 mmol/L) dose-dependently diminished migration velocity or even blocked migration of CSD. The CSD blockade by the alpha(2) antagonist yohimbine (1.75 mmol/L) was because of its action on inhibitory 5 HT(1A) receptors. None of the substances in the concentrations used had influence on regional cerebral blood flow or on systemic arterial blood pressure. The data suggest that the interference of these compounds with CSD may contribute to their beneficial therapeutic effect. The effect of beta-receptor antagonists in human migraine needs further exploration, since these drugs also work in migraine without aura. PMID- 15829917 TI - Inhibition of angiogenic properties of brain endothelial cells by platelet derived sphingosine-1-phosphate. AB - The platelet-derived lysophospholipid sphingosine-1-phosphate (S1P) is present in blood plasma and is one of the most potent growth factors displaying proangiogenic activity towards endothelial cells (EC) derived from various tissues. The paracrine regulation of brain angiogenesis by platelet-derived growth factors is, however, poorly understood. In the present study, we assessed the role of S1P on brain EC migration and tubulogenesis, using rat brain-derived (RBE4) EC as an in vitro model. We show that S1P inhibits brain EC migration and tubulogenesis, while it displays proangiogenic activity towards noncerebral EC. Overexpression of the S1P receptor S1P-1 in RBE4 cells potentiated all of the S1P mediated events. We also show that the lack of expression of MT1-MMP, a membrane bound matrix metalloproteinase that is thought to cooperate with S1P in tubulogenic processes, may explain the antiangiogenic activity of S1P on brain vasculature. Altogether our results support the hypothesis of a tissue-specific, antiangiogenic role of S1P in the brain, which may help to stabilize the cerebral vasculature and thus have crucial impact on the setting and regulation of normal brain vascularization. PMID- 15829918 TI - VEGF protects brain against focal ischemia without increasing blood--brain permeability when administered intracerebroventricularly. AB - Delayed administration of vascular endothelial growth factor (VEGF) promotes functional recovery after focal cerebral ischemia. However, early intravenous injection of VEGF increases blood-brain barrier (BBB) leakage, hemorrhagic transformation and infarct volume whereas its application to cortical surface is neuroprotective. We have investigated whether or not early intracerebroventricular administration of VEGF could replicate the neuroprotective effect observed with topical application and the mechanism of action of this protection. Mice were subjected to 90 mins middle cerebral artery (MCA) occlusion and 24 h of reperfusion. Vascular endothelial growth factor (8 ng, intracerebroventricular) was administered 1 or 3 h after reperfusion. Compared with the vehicle-treated (intracerebroventricular) group, VEGF decreased the infarct volume along with BBB leakage in both treatment groups. Neurologic disability scores improved in parallel to the changes in infarct volume. Independently of the decrease in infarct size, VEGF also reduced the number of TUNEL-positive apoptotic neurons. Phospo-Akt levels were significantly higher in ischemic hemispheres of the VEGF-treated mice. Contrary to intracerebroventricular route, intravenous administration of VEGF (15 microg/kg) enhanced the infarct volume as previously reported for the rat. In conclusion, single intracerebroventricular injection of VEGF protects brain against ischemia without adversely affecting BBB permeability, and has a relatively long therapeutic time window. This early neuroprotective action, observed well before recovery-promoting actions such as angiogenesis, possibly involves activation of the PI-3-Akt pathway. PMID- 15829919 TI - Fast vesicle replenishment allows indefatigable signalling at the first auditory synapse. AB - Ribbon-type synapses in inner hair cells of the mammalian cochlea encode the complexity of auditory signals by fast and tonic release through fusion of neurotransmitter-containing vesicles. At any instant, only about 100 vesicles are tethered to the synaptic ribbon, and about 14 of these are docked to the plasma membrane, constituting the readily releasable pool. Although this pool contains about the same number of vesicles as that of conventional synapses, ribbon release sites operate at rates of about two orders of magnitude higher and with submillisecond precision. How these sites replenish their vesicles so efficiently remains unclear. We show here, using two-photon imaging of single release sites in the intact cochlea, that preformed vesicles derived from cytoplasmic vesicle generating compartments participate in fast release and replenishment. Vesicles were released at a maximal initial rate of 3 per millisecond during a depolarizing pulse, and were replenished at a rate of 1.9 per millisecond. We propose that such rapid resupply of vesicles enables temporally precise and sustained release rates. This may explain how the first auditory synapse can encode with indefatigable precision without having to rely on the slow, local endocytic vesicle cycle. PMID- 15829920 TI - Cellular APOBEC3G restricts HIV-1 infection in resting CD4+ T cells. AB - In contrast to activated CD4+ T cells, resting human CD4+ T cells circulating in blood are highly resistant to infection with human immunodeficiency virus (HIV). Whether the inability of HIV to infect these resting CD4+ T cells is due to the lack of a key factor, or alternatively reflects the presence of an efficient mechanism for defence against HIV, is not clear. Here we show that the anti retroviral deoxycytidine deaminase APOBEC3G strongly protects unstimulated peripheral blood CD4+ T cells against HIV-1 infection. In activated CD4+ T cells, cytoplasmic APOBEC3G resides in an enzymatically inactive, high-molecular-mass (HMM) ribonucleoprotein complex that converts to an enzymatically active low molecular-mass (LMM) form after treatment with RNase. In contrast, LMM APOBEC3G predominates in unstimulated CD4+ T cells, where HIV-1 replication is blocked and reverse transcription is impaired. Mitogen activation induces the recruitment of LMM APOBEC3G into the HMM complex, and this correlates with a sharp increase in permissivity for HIV infection in these stimulated cells. Notably, when APOBEC3G specific small interfering RNAs are introduced into unstimulated CD4+ T cells, the early replication block encountered by HIV-1 is greatly relieved. Thus, LMM APOBEC3G functions as a potent post-entry restriction factor for HIV-1 in unstimulated CD4+ T cells. Surprisingly, sequencing of the reverse transcripts slowly formed in unstimulated CD4+ T cells reveals only low levels of dG dA hypermutation, raising the possibility that the APOBEC3G-restricting activity may not be strictly dependent on deoxycytidine deamination PMID- 15829921 TI - Don't rely on Uncle Sam. PMID- 15829922 TI - US health officials rally behind bid to relax rules on embryo research. PMID- 15829923 TI - Divergent local laws threaten to stifle Europe's stem-cell project. PMID- 15829924 TI - Law change imperils studies of ancient human remains. PMID- 15829925 TI - Vaccination will work better than culling, say bird flu experts. PMID- 15829926 TI - Ideas abound as Japan aims to boost its space image. PMID- 15829927 TI - Shuttle reports for duty...despite the risks. PMID- 15829928 TI - Gene therapists urged to learn more immunology. PMID- 15829929 TI - Health study sets sights on a million people. PMID- 15829930 TI - War of words deepens divide over biodefence funds. PMID- 15829931 TI - Palestinian unease sparks fresh calls for Israeli boycott. PMID- 15829932 TI - Study gives backing to HIV drug for mothers. PMID- 15829933 TI - Climate science: the dustiest place on Earth. PMID- 15829934 TI - Physics and the public: science as illusion. PMID- 15829935 TI - NIH conflicts rules are not right for universities. PMID- 15829936 TI - Changes in China call for new health solutions. PMID- 15829937 TI - Influenza drug could abort a pandemic. PMID- 15829942 TI - Flight of fancy. PMID- 15829943 TI - Medicine: aborting the birth of cancer. PMID- 15829944 TI - Climate change: water cycle shifts gear. PMID- 15829945 TI - Evolution: warm-hearted crocs. PMID- 15829946 TI - Geophysics: double-crossed again. PMID- 15829947 TI - Neurobiology: channels for pathfinding. PMID- 15829949 TI - Astrophysics: two's company. PMID- 15829951 TI - Greenhouse gases: low methane leakage from gas pipelines. AB - Using natural gas for fuel releases less carbon dioxide per unit of energy produced than burning oil or coal, but its production and transport are accompanied by emissions of methane, which is a much more potent greenhouse gas than carbon dioxide in the short term. This calls into question whether climate forcing could be reduced by switching from coal and oil to natural gas. We have made measurements in Russia along the world's largest gas-transport system and find that methane leakage is in the region of 1.4%, which is considerably less than expected and comparable to that from systems in the United States. Our calculations indicate that using natural gas in preference to other fossil fuels could be useful in the short term for mitigating climate change. PMID- 15829952 TI - Planetary science: constant illumination at the lunar north pole. AB - Images returned by the spacecraft Clementine have been used to produce a quantitative illumination map of the north pole of the Moon, revealing the percentage of time that points on the surface are illuminated during the lunar day. We have used this map to identify areas that are constantly illuminated during a lunar day in summer and which may therefore be in permanent sunlight. All are located on the northern rim of Peary crater, close to the north pole. Permanently sunlit areas represent prime locations for lunar outpost sites as they have abundant solar energy, are relatively benign thermally (when compared with equatorial regions), and are close to permanently shadowed regions that may contain water ice. PMID- 15829953 TI - Wnt signalling in stem cells and cancer. AB - The canonical Wnt cascade has emerged as a critical regulator of stem cells. In many tissues, activation of Wnt signalling has also been associated with cancer. This has raised the possibility that the tightly regulated self-renewal mediated by Wnt signalling in stem and progenitor cells is subverted in cancer cells to allow malignant proliferation. Insights gained from understanding how the Wnt pathway is integrally involved in both stem cell and cancer cell maintenance and growth in the intestinal, epidermal and haematopoietic systems may serve as a paradigm for understanding the dual nature of self-renewal signals. PMID- 15829954 TI - Lead isotopes reveal bilateral asymmetry and vertical continuity in the Hawaiian mantle plume. AB - The two parallel chains of Hawaiian volcanoes ('Loa' and 'Kea') are known to have statistically different but overlapping radiogenic isotope characteristics. This has been explained by a model of a concentrically zoned mantle plume, where the Kea chain preferentially samples a more peripheral portion of the plume. Using high-precision lead isotope data for both centrally and peripherally located volcanoes, we show here that the two trends have very little compositional overlap and instead reveal bilateral, non-concentric plume zones, probably derived from the plume source in the mantle. On a smaller scale, along the Kea chain, there are isotopic differences between the youngest lavas from the Mauna Kea and Kilauea volcanoes, but the 550-thousand-year-old Mauna Kea lavas are isotopically identical to Kilauea lavas, consistent with Mauna Kea's position relative to the plume, which was then similar to that of present-day Kilauea. We therefore conclude that narrow (less than 50 kilometres wide) compositional streaks, as well as the larger-scale bilateral zonation, are vertically continuous over tens to hundreds of kilometres within the plume. PMID- 15829955 TI - A common sex-dependent mutation in a RET enhancer underlies Hirschsprung disease risk. AB - The identification of common variants that contribute to the genesis of human inherited disorders remains a significant challenge. Hirschsprung disease (HSCR) is a multifactorial, non-mendelian disorder in which rare high-penetrance coding sequence mutations in the receptor tyrosine kinase RET contribute to risk in combination with mutations at other genes. We have used family-based association studies to identify a disease interval, and integrated this with comparative and functional genomic analysis to prioritize conserved and functional elements within which mutations can be sought. We now show that a common non-coding RET variant within a conserved enhancer-like sequence in intron 1 is significantly associated with HSCR susceptibility and makes a 20-fold greater contribution to risk than rare alleles do. This mutation reduces in vitro enhancer activity markedly, has low penetrance, has different genetic effects in males and females, and explains several features of the complex inheritance pattern of HSCR. Thus, common low-penetrance variants, identified by association studies, can underlie both common and rare diseases. PMID- 15829956 TI - DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. AB - During the evolution of cancer, the incipient tumour experiences 'oncogenic stress', which evokes a counter-response to eliminate such hazardous cells. However, the nature of this stress remains elusive, as does the inducible anti cancer barrier that elicits growth arrest or cell death. Here we show that in clinical specimens from different stages of human tumours of the urinary bladder, breast, lung and colon, the early precursor lesions (but not normal tissues) commonly express markers of an activated DNA damage response. These include phosphorylated kinases ATM and Chk2, and phosphorylated histone H2AX and p53. Similar checkpoint responses were induced in cultured cells upon expression of different oncogenes that deregulate DNA replication. Together with genetic analyses, including a genome-wide assessment of allelic imbalances, our data indicate that early in tumorigenesis (before genomic instability and malignant conversion), human cells activate an ATR/ATM-regulated DNA damage response network that delays or prevents cancer. Mutations compromising this checkpoint, including defects in the ATM-Chk2-p53 pathway, might allow cell proliferation, survival, increased genomic instability and tumour progression. PMID- 15829957 TI - Nucleosynthetic signatures of the first stars. AB - The chemically most primitive stars provide constraints on the nature of the first stellar objects that formed in the Universe; elements other than hydrogen, helium and traces of lithium present within these objects were generated by nucleosynthesis in the very first stars. The relative abundances of elements in the surviving primitive stars reflect the masses of the first stars, because the pathways of nucleosynthesis are quite sensitive to stellar masses. Several models have been suggested to explain the origin of the abundance pattern of the giant star HE0107-5240, which hitherto exhibited the highest deficiency of heavy elements known. Here we report the discovery of HE1327-2326, a subgiant or main sequence star with an iron abundance about a factor of two lower than that of HE0107-5240. Both stars show extreme overabundances of carbon and nitrogen with respect to iron, suggesting a similar origin of the abundance patterns. The unexpectedly low Li and high Sr abundances of HE1327-2326, however, challenge existing theoretical understanding: no model predicts the high Sr abundance or provides a Li depletion mechanism consistent with data available for the most metal-poor stars. PMID- 15829958 TI - Planet-planet scattering in the upsilon Andromedae system. AB - Doppler spectroscopy has detected 152 planets around nearby stars. A major puzzle is why many of their orbits are highly eccentric; all planets in our Solar System are on nearly circular orbits, as is expected if they formed by accretion processes in a protostellar disk. Several mechanisms have been proposed to generate large eccentricities after planet formation, but so far there has been little observational evidence to support any particular model. Here we report that the current orbital configuration of the three giant planets around upsilon Andromedae (upsilon And) probably results from a close dynamical interaction with another planet, now lost from the system. The planets started on nearly circular orbits, but chaotic evolution caused the outer planet (upsilon And d) to be perturbed suddenly into a higher-eccentricity orbit. The coupled evolution of the system then causes slow periodic variations in the eccentricity of the middle planet (upsilon And c). Indeed, we show that upsilon And c periodically returns to a very nearly circular state every 6,700 years. PMID- 15829959 TI - Sensitivity gains in chemosensing by lasing action in organic polymers. AB - Societal needs for greater security require dramatic improvements in the sensitivity of chemical and biological sensors. To meet this challenge, increasing emphasis in analytical science has been directed towards materials and devices having highly nonlinear characteristics; semiconducting organic polymers (SOPs), with their facile excited state (exciton) transport, are prime examples of amplifying materials. SOPs have also been recognized as promising lasing materials, although the susceptibility of these materials to optical damage has thus far limited applications. Here we report that attenuated lasing in optically pumped SOP thin films displays a sensitivity to vapours of explosives more than 30 times higher than is observed from spontaneous emission. Critical to this achievement was the development of a transducing polymer with high thin-film quantum yield, a high optical damage threshold in ambient atmosphere and a record low lasing threshold. Trace vapours of the explosives 2,4,6-trinitrotoluene (TNT) and 2,4-dinitrotoluene (DNT) introduce non-radiative deactivation pathways that compete with stimulated emission. We demonstrate that the induced cessation of the lasing action, and associated sensitivity enhancement, is most pronounced when films are pumped at intensities near their lasing threshold. The combined gains from amplifying materials and lasing promise to deliver sensors that can detect explosives with unparalleled sensitivity. PMID- 15829960 TI - Light-induced shape-memory polymers. AB - Materials are said to show a shape-memory effect if they can be deformed and fixed into a temporary shape, and recover their original, permanent shape only on exposure to an external stimulus. Shape-memory polymers have received increasing attention because of their scientific and technological significance. In principle, a thermally induced shape-memory effect can be activated by an increase in temperature (also obtained by heating on exposure to an electrical current or light illumination). Several papers have described light-induced changes in the shape of polymers and gels, such as contraction, bending or volume changes. Here we report that polymers containing cinnamic groups can be deformed and fixed into pre-determined shapes--such as (but not exclusively) elongated films and tubes, arches or spirals--by ultraviolet light illumination. These new shapes are stable for long time periods, even when heated to 50 degrees C, and they can recover their original shape at ambient temperatures when exposed to ultraviolet light of a different wavelength. The ability of polymers to form different pre-determined temporary shapes and subsequently recover their original shape at ambient temperatures by remote light activation could lead to a variety of potential medical and other applications. PMID- 15829961 TI - A doubling of the post-perovskite phase boundary and structure of the Earth's lowermost mantle. AB - The thermal structure of the Earth's lowermost mantle--the D'' layer spanning depths of approximately 2,600-2,900 kilometres--is key to understanding the dynamical state and history of our planet. Earth's temperature profile (the geotherm) is mostly constrained by phase transitions, such as freezing at the inner-core boundary or changes in crystal structure within the solid mantle, that are detected as discontinuities in seismic wave speed and for which the pressure and temperature conditions can be constrained by experiment and theory. A recently discovered phase transition at pressures of the D'' layer is ideally situated to reveal the thermal structure of the lowermost mantle, where no phase transitions were previously known to exist. Here we show that a pair of seismic discontinuities observed in some regions of D'' can be explained by the same phase transition as the result of a double-crossing of the phase boundary by the geotherm at two different depths. This simple model can also explain why a seismic discontinuity is not observed in some other regions, and provides new constraints for the magnitude of temperature variations within D''. PMID- 15829962 TI - Permian tetrapods from the Sahara show climate-controlled endemism in Pangaea. AB - New fossils from the Upper Permian Moradi Formation of northern Niger provide an insight into the faunas that inhabited low-latitude, xeric environments near the end of the Palaeozoic era (approximately 251 million years ago). We describe here two new temnospondyl amphibians, the cochleosaurid Nigerpeton ricqlesi gen. et sp. nov. and the stem edopoid Saharastega moradiensis gen. et sp. nov., as relicts of Carboniferous lineages that diverged 40-90 million years earlier. Coupled with a scarcity of therapsids, the new finds suggest that faunas from the poorly sampled xeric belt that straddled the Equator during the Permian period differed markedly from well-sampled faunas that dominated tropical-to-temperate zones to the north and south. Our results show that long-standing theories of Late Permian faunal homogeneity are probably oversimplified as the result of uneven latitudinal sampling. PMID- 15829963 TI - Hair cell synaptic ribbons are essential for synchronous auditory signalling. AB - Hearing relies on faithful synaptic transmission at the ribbon synapse of cochlear inner hair cells (IHCs). At present, the function of presynaptic ribbons at these synapses is still largely unknown. Here we show that anchoring of IHC ribbons is impaired in mouse mutants for the presynaptic scaffolding protein Bassoon. The lack of active-zone-anchored synaptic ribbons reduced the presynaptic readily releasable vesicle pool, and impaired synchronous auditory signalling as revealed by recordings of exocytic IHC capacitance changes and sound-evoked activation of spiral ganglion neurons. Both exocytosis of the hair cell releasable vesicle pool and the number of synchronously activated spiral ganglion neurons co-varied with the number of anchored ribbons during development. Interestingly, ribbon-deficient IHCs were still capable of sustained exocytosis with normal Ca2+-dependence. Endocytic membrane retrieval was intact, but an accumulation of tubular and cisternal membrane profiles was observed in ribbon-deficient IHCs. We conclude that ribbon-dependent synchronous release of multiple vesicles at the hair cell afferent synapse is essential for normal hearing. PMID- 15829964 TI - Sheep retrovirus structural protein induces lung tumours. AB - Jaagsiekte sheep retrovirus (JSRV) causes a contagious lung cancer in sheep and goats, with significant animal health and economic consequences. The host range of JSRV is in part limited by species-specific differences in the virus entry receptor, hyaluronidase 2 (Hyal2), which is not functional as a receptor in mice but is functional in humans. Sheep are immunotolerant of JSRV because of the expression of closely related endogenous retroviruses, which are not present in humans and most other species, and this may facilitate oncogenesis. Here we show that expression of the JSRV envelope (Env) protein alone in lungs of mice, by using a replication-incompetent adeno-associated virus vector, results in tumours with a bronchiolo-alveolar localization like those seen in sheep. Whereas lethal disease was observed in immunodeficient mice, tumour development was almost entirely blocked in immunocompetent mice. Our results provide a rare example of an oncogenic viral structural protein, show that interaction of the viral Env protein with the virus entry receptor Hyal2 is not required for tumorigenesis, and indicate that immune recognition of Env can protect against JSRV tumorigenesis. PMID- 15829965 TI - Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. AB - DNA damage checkpoint genes, such as p53, are frequently mutated in human cancer, but the selective pressure for their inactivation remains elusive. We analysed a panel of human lung hyperplasias, all of which retained wild-type p53 genes and had no signs of gross chromosomal instability, and found signs of a DNA damage response, including histone H2AX and Chk2 phosphorylation, p53 accumulation, focal staining of p53 binding protein 1 (53BP1) and apoptosis. Progression to carcinoma was associated with p53 or 53BP1 inactivation and decreased apoptosis. A DNA damage response was also observed in dysplastic nevi and in human skin xenografts, in which hyperplasia was induced by overexpression of growth factors. Both lung and experimentally-induced skin hyperplasias showed allelic imbalance at loci that are prone to DNA double-strand break formation when DNA replication is compromised (common fragile sites). We propose that, from its earliest stages, cancer development is associated with DNA replication stress, which leads to DNA double-strand breaks, genomic instability and selective pressure for p53 mutations. PMID- 15829966 TI - Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. AB - Poly(ADP-ribose) polymerase (PARP1) facilitates DNA repair by binding to DNA breaks and attracting DNA repair proteins to the site of damage. Nevertheless, PARP1-/- mice are viable, fertile and do not develop early onset tumours. Here, we show that PARP inhibitors trigger gamma-H2AX and RAD51 foci formation. We propose that, in the absence of PARP1, spontaneous single-strand breaks collapse replication forks and trigger homologous recombination for repair. Furthermore, we show that BRCA2-deficient cells, as a result of their deficiency in homologous recombination, are acutely sensitive to PARP inhibitors, presumably because resultant collapsed replication forks are no longer repaired. Thus, PARP1 activity is essential in homologous recombination-deficient BRCA2 mutant cells. We exploit this requirement in order to kill BRCA2-deficient tumours by PARP inhibition alone. Treatment with PARP inhibitors is likely to be highly tumour specific, because only the tumours (which are BRCA2-/-) in BRCA2+/- patients are defective in homologous recombination. The use of an inhibitor of a DNA repair enzyme alone to selectively kill a tumour, in the absence of an exogenous DNA damaging agent, represents a new concept in cancer treatment. PMID- 15829967 TI - Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. AB - BRCA1 and BRCA2 are important for DNA double-strand break repair by homologous recombination, and mutations in these genes predispose to breast and other cancers. Poly(ADP-ribose) polymerase (PARP) is an enzyme involved in base excision repair, a key pathway in the repair of DNA single-strand breaks. We show here that BRCA1 or BRCA2 dysfunction unexpectedly and profoundly sensitizes cells to the inhibition of PARP enzymatic activity, resulting in chromosomal instability, cell cycle arrest and subsequent apoptosis. This seems to be because the inhibition of PARP leads to the persistence of DNA lesions normally repaired by homologous recombination. These results illustrate how different pathways cooperate to repair damage, and suggest that the targeted inhibition of particular DNA repair pathways may allow the design of specific and less toxic therapies for cancer. PMID- 15829968 TI - Transcriptional regulation of a metastasis suppressor gene by Tip60 and beta catenin complexes. AB - Defining the molecular strategies that integrate diverse signalling pathways in the expression of specific gene programmes that are critical in homeostasis and disease remains a central issue in biology. This is particularly pertinent in cancer biology because downregulation of tumour metastasis suppressor genes is a common occurrence, and the underlying molecular mechanisms are not well established. Here we report that the downregulation of a metastasis suppressor gene, KAI1, in prostate cancer cells involves the inhibitory actions of beta catenin, along with a reptin chromatin remodelling complex. This inhibitory function of beta-catenin-reptin requires both increased beta-catenin expression and recruitment of histone deacetylase activity. The coordinated actions of beta catenin-reptin components that mediate the repressive state serve to antagonize a Tip60 coactivator complex that is required for activation; the balance of these opposing complexes controls the expression of KAI1 and metastatic potential. The molecular mechanisms underlying the antagonistic regulation of beta-catenin reptin and the Tip60 coactivator complexes for the metastasis suppressor gene, KAI1, are likely to be prototypic of a selective downregulation strategy for many genes, including a subset of NF-kappaB target genes. PMID- 15829969 TI - Structure of the apoptotic protease-activating factor 1 bound to ADP. AB - Apoptosis is executed by caspases, which undergo proteolytic activation in response to cell death stimuli. The apoptotic protease-activating factor 1 (Apaf 1) controls caspase activation downstream of mitochondria. During apoptosis, Apaf 1 binds to cytochrome c and in the presence of ATP/dATP forms an apoptosome, leading to the recruitment and activation of the initiator caspase, caspase-9 (ref. 2). The mechanisms underlying Apaf-1 function are largely unknown. Here we report the 2.2-A crystal structure of an ADP-bound, WD40-deleted Apaf-1, which reveals the molecular mechanism by which Apaf-1 exists in an inactive state before ATP binding. The amino-terminal caspase recruitment domain packs against a three-layered alpha/beta fold, a short helical motif and a winged-helix domain, resulting in the burial of the caspase-9-binding interface. The deeply buried ADP molecule serves as an organizing centre to strengthen interactions between these four adjoining domains, thus locking Apaf-1 in an inactive conformation. Apaf-1 binds to and hydrolyses ATP/dATP and their analogues. The binding and hydrolysis of nucleotides seem to drive conformational changes that are essential for the formation of the apoptosome and the activation of caspase-9. PMID- 15829970 TI - Out in the cold. PMID- 15829971 TI - High-energy career lines. PMID- 15829973 TI - Recruiters and industry. Microsoft's European perspective. PMID- 15829976 TI - Epstein-Barr virus latent membrane protein 1 represses p53-mediated DNA repair and transcriptional activity. AB - The latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV), a viral oncogene, is essential for transformation of resting B cells by the virus. We previously demonstrated that LMP1 could repress DNA repair in p53-wild-type and p53-deficient human epithelial cells. In this study, using a host cell reactivation (HCR) assay, we demonstrated that p53-enhanced DNA repair was repressed by LMP1 in p53-deficient cells. Moreover, we found that LMP1 was able to repress p53-dependent transcriptional activity. Regarding the mechanisms of p53 repression by LMP1, we found that LMP1 did not inhibit p53 function through direct interaction, by promoting protein degradation or reducing its DNA-binding ability. Using chimeric proteins in the reporter assay, we demonstrated that LMP1 inhibited p53 transactivation by influencing the N-terminal transactivation domain of p53. Subsequent experiments using various LMP1 deletion mutants indicated that a C-terminus-activating region of LMP1, CTAR1 or CTAR2, is responsible for the repression of p53-mediated DNA repair and p53-dependent transcription, which is correlated with the region responsible for NF-kappaB activation. Furthermore, blockage of NF-kappaB signalling by IkappaB-DeltaN was shown to abolish the repression of p53 by LMP1, suggesting that LMP1 likely repressed p53 function through the NF-kappaB pathway. Based on these results, we propose that inhibition of p53-dependent transcriptional activity and DNA repair by LMP1 results in the loss of p53 activity for maintaining genomic stability, which may contribute to the oncogenesis of LMP1 in human epithelial cells. PMID- 15829977 TI - A 5'-distal enhanceosome in the PDGF-A gene is activated in choriocarcinoma cells via ligand-independent binding of vitamin D receptor and constitutive jun kinase signaling. AB - Overexpression of platelet-derived growth factor A-chain (PDGF-A) is clearly linked to autocrine and paracrine stimulation of malignant growth in many human cancers. We have shown previously that PDGF-A overexpression in choriocarcinoma, hepatoma and lung carcinoma cell lines is driven by the activity of a 66 bp enhancer element (ACE66) located approximately 7 kb upstream of the PDGF-A transcription start site. In this study, the ACE66 element is shown to be activated in JEG-3 choriocarcinoma cells through synergistic interactions between consensus DNA motifs for binding of vitamin D receptor, AP1 and ELK1. Binding of the vitamin D/retinoid-X receptor (VDR/RXRalpha) heterodimer to the ACE66 element was reconstituted in vitro with recombinant VDR/RXRalpha and with JEG-3 nuclear extract, and was verified in living JEG-3 cells by chromatin immunoprecipitation analysis. Transcriptional activity of the ACE66 element, as well as occupancy of the element by VDR/RXRalpha, was shown to be independent of stimulation with the hormonal VDR ligand, 1,25-dihydroxyvitamin D3. The jun kinase pathway of mitogen activated protein kinase (MAPK) signaling was shown to activate the ACE66 enhancer, most likely through activation of factors binding to the AP1 element. These results identify a novel mechanism of transcriptional enhancement involving ligand-independent activity of the VDR/RXR heterodimer and MAPK signaling pathways that appears to play an important role in the overexpression of PDGF in many different settings of human malignancy. PMID- 15829978 TI - Functional correlates of mutation of the Asp32 and Gly34 residues of beta catenin. AB - Beta-catenin is a multifunctional protein involved in both cadherin-mediated adhesion and the wnt signaling cascade. Mutations in exon 3 of beta-catenin have been identified in many cancers. In addition to disruption of key serine and threonine residues, mutations are frequently reported in other residues in exon 3 that are not kinase substrates. The most frequently mutated nonserine/threonine residues are D32 and G34. Since D32 and G34 are part of the ubiquitination destruction motif, DSGPhiXS, we hypothesize that this motif may contribute to disruption of beta-catenin homeostasis and lead to cellular transformation. We demonstrate that the mutants D32A and G34A exhibit no change in phosphorylation by GSK3beta, but display reduced ubiquitination compared to wild-type and S33A mutant beta-catenin. To assess the functional implications of these mutations, we created stable MDCK cell lines expressing these constructs. We found that stable cell lines harboring D32A-mutated beta-catenin were highly transformed, while S33A and G34 demonstrated only weak transforming properties in our assays. Despite altered ubiquitination status and increased transformation, the D32A mutant cell line does not display transcriptional activation of standard target genes. Therefore, D32A mutation may mediate transformation by an alternative beta catenin-mediated signaling pathway. PMID- 15829979 TI - Definition and characterization of a region of 1p36.3 consistently deleted in neuroblastoma. AB - Substantial genomic and functional evidence from primary tumors and cell lines indicates that a consistent region of distal chromosome 1p is deleted in a sizable proportion of human neuroblastomas, suggesting that this region contains one or more tumor suppressor genes. To determine systematically and precisely the location and extent of 1p deletion in neuroblastomas, we performed allelic loss studies of 737 primary neuroblastomas and genotype analysis of 46 neuroblastoma cell lines. Together, the results defined a single region within 1p36.3 that was consistently deleted in 25% of tumors and 87% of cell lines. Two neuroblastoma patients had constitutional deletions of distal 1p36 that overlapped the tumor defined region. The tumor- and constitutionally-derived deletions together defined a smallest region of consistent deletion (SRD) between D1S2795 and D1S253. The 1p36.3 SRD was deleted in all but one of the 184 tumors with 1p deletion. Physical mapping and DNA sequencing determined that the SRD minimally spans an estimated 729 kb. Genomic content and sequence analysis of the SRD identified 15 characterized, nine uncharacterized, and six predicted genes in the region. The RNA expression profiles of 21 of the genes were investigated in a variety of normal tissues. The SHREW1 and KCNAB2 genes both had tissue-restricted expression patterns, including expression in the nervous system. In addition, a novel gene (CHD5) with strong homology to proteins involved in chromatin remodeling was expressed mainly in neural tissues. Together, these results suggest that one or more genes involved in neuroblastoma tumorigenesis or tumor progression are likely contained within this region. PMID- 15829980 TI - Deregulated expression of RasGRP1 initiates thymic lymphomagenesis independently of T-cell receptors. AB - RasGRP1 is a Ras-specific exchange factor, which is activated by T-cell receptor (TCR) and promotes TCR-dependent positive selection of thymocytes. RasGRP1 is highly expressed on most T lymphocytic leukemias and is a common site of proviral insertion in retrovirus-induced murine T-cell lymphomas. We used RasGRP1 transgenic mice to determine if deregulated expression of RasGRP1 has a causative role in the development of T-cell malignancies. Thymic lymphomas occurred in three different RasGRP1 transgenic mouse lines. Thymocyte transformation correlated with high transgene expression in early stage lymphomas, indicating that deregulated RasGRP1 expression contributed to the initiation of lymphomagenesis. Expression of the positively selectable H-Y TCR accelerated lymphomagenesis in RasGRP1 transgenic mice. However, the transformed thymocytes lacked markers of positive selection and lymphomas occurred when positive selection was precluded by negative selection of the H-Y TCR. Therefore, initiation of lymphomagenesis via RasGRP1 was not associated with TCR-dependent positive selection of thymocytes. Thymic lymphomas occurred in RasGRP1 transgenic/Rag2-/- mice, demonstrating that neither TCR nor pre-TCR were required for RasGRP1-driven lymphomagenesis. The RasGRP1 transgene conferred pre-TCR independent survival and proliferation of immature thymocytes, suggesting that deregulated expression of RasGRP1 promotes lymphomagenesis by expanding the pool of thymocytes which are susceptible to transformation. PMID- 15829982 TI - Split genes uncovered through science fusion. PMID- 15829981 TI - Cyclin A1, the alternative A-type cyclin, contributes to G1/S cell cycle progression in somatic cells. AB - Cyclin A1 is an alternative A-type cyclin that is essential for spermatogenesis, but it is also expressed in hematopoietic progenitor cells and in acute myeloid leukemia. Its functions during cell cycle progression of somatic cells are incompletely understood. Here, we have analysed the cell cycle functions of cyclin A1 in transformed and nontransformed cells. Murine embryonic fibroblasts derived from cyclin A1-deficient mice were significantly impaired in their proliferative capacity. In accordance, cyclin A1-/- cells accumulated in G1 and G2/M phase while the percentage of S phase cells decreased. Also, lectin stimulated splenic lymphocytes from cyclin A1-/- mice proliferated slower than their wild-type counterparts. Forced cyclin A1 overexpression in NIH3T3 cells and in U937 leukemic cells either by transient transfection or by retroviral infection enhanced S phase entry. Consequently, siRNA mediated silencing of cyclin A1 in highly cyclin A1 expressing ML1 leukemic cells significantly slowed S phase entry, decreased proliferation and inhibited colony formation. Taken together, these analyses demonstrate that cyclin A1 contributes to G1 to S cell cycle progression in somatic cells. Cyclin A1 overexpression enhances S phase entry consistent with an oncogenic function. Finally, cyclin A1 might be a therapeutic target since its silencing inhibited leukemia cell growth. PMID- 15829983 TI - Quantitative evaluation of the petal shape variation in Primula sieboldii caused by breeding process in the last 300 years. AB - Primula sieboldii: (E. Morren) has been a popular garden plant at least since the Edo period, about 300 years ago. We compared petal form between cultivars and wild populations in order to characterise the changes that have occurred during domestication. The comparison was made using EF-PCA analysis, which describes overall petal shape mathematically by transforming petal contour coordinates into elliptic Fourier descriptors; it subsequently summarises these descriptors by principal component analysis (PCA). Rearing cultivars in a common-garden experiment identified the PCs with a substantial genetic element. A clear heritable component was detected for the PCs characterising symmetrical variation in flower shape, but not the asymmetrical variation. Wild populations of this species have become endangered owing to habitat destruction by human activity, and many lowland floodplain habitats have been lost. Variation within the remaining wild populations was significantly lower than in the cultivars for PC1 (aspect ratio), PC3 (curvature of proximal and distal parts) and petal area; but not for PC2 (depth of head notch) and PC4 (position of the centre of gravity). The shifts in petal form from the wild populations to the cultivars parallel those seen in other crop-types following domestication, including an increase in size and diversity of forms: cultivars have shallower head notches, more fan shaped petals and larger petals than do wild P. sieboldii. PMID- 15829984 TI - Evidence for paternal transmission and heteroplasmy in the mitochondrial genome of Silene vulgaris, a gynodioecious plant. AB - Gynodioecy refers to the co-occurrence of females and hermaphrodites in the same population. In many gynodioecious plants, sex is determined by an epistatic interaction between mitochondrial and nuclear genes, resulting in intragenomic evolutionary conflict, should the mitochondrial genome be maternally inherited. While maternal inheritance of the mitochondrial genome is common in angiosperms, few gynodioecious species have been studied. Here, the inheritance of the mitochondrial genes atpA and coxI was studied in 318 Silene vulgaris individuals distributed among 23 crosses. While maternal inheritance was indicated in 96% of the individuals studied, one or more individuals from each of four sib groups displayed a genotype that was identical to the father, or that did not match either parent. Given evidence that inheritance is not strictly maternal, it was hypothesized that some individuals could carry a mixture of maternally and paternally derived copies of the mitochondrial genome, a condition known as heteroplasmy. Since heteroplasmy might be difficult to detect should multiple versions of the mitochondrial genome co-occur in highly unequal copy number, a method was devised to amplify low-copy number forms of atpA differentially. Evidence for heteroplasmy was found in 23 of the 99 individuals studied, including cases in which the otherwise cryptic form of atpA matched the paternal genotype. The distribution of shared nucleotide sequence polymorphism among atpA haplotypes and the results of a population survey of the joint distribution of atpA and coxI haplotypes across individuals supports the hypothesis that heteroplasmy facilitates formation of novel mitochondrial genotypes by recombination. PMID- 15829985 TI - Multinucleate storage cells in Douglas-fir (Pseudotsuga menziesii (Mirbel) Franco) and the effect of seed parasitism by the chalcid Megastigmus spermotrophus Wachtl. AB - Megagametophytes of Douglas-fir (Pseudotsuga menziesii (Mirbel) Franco) accumulated storage products following fertilization. As megagametophytes matured, the number of nuclei per cell rose, resulting in syncytial storage cells. Studies carried out on trees in France and Canada confirmed that such previously unreported, free nuclear cells were a normal part of late megagametophyte development. Unfertilized megagametophytes showed that some binucleate cells before degeneration resulted in empty seed. Insect parasitism prevented megagametophyte abortion in unfertilized ovules. Oviposition by a torymid chalcid wasp (Megastigmus spermotrophus Wachtl) early in megagametophyte development resulted in normal megagametophyte development. Around the time of plant egg maturation, binucleate and trinucleate cells were observed. As megagametophytes matured, multinucleate mature storage cells rich in proteins, lipids and starch were formed. The insect was able to induce identical nuclear behaviour in infested, unfertilized megagametophytes, as that of uninfested, fertilized megagametophytes. PMID- 15829986 TI - Reconciling nuclear microsatellite and mitochondrial marker estimates of population structure: breeding population structure of Chesapeake Bay striped bass (Morone saxatilis). AB - Comparative analyses of nuclear and organelle genetic markers may help delineate evolutionarily significant units or management units, although population differentiation estimates from multiple genomes can also conflict. Striped bass (Morone saxatilis) are long-lived, highly migratory anadromous fish recently recovered from a severe decline in population size. Previous studies with protein, nuclear DNA and mitochondrial DNA (mtDNA) markers produced discordant results, and it remains uncertain if the multiple tributaries within Chesapeake Bay constitute distinct management units. Here, 196 young-of-the-year (YOY) striped bass were sampled from Maryland's Choptank, Potomac and Nanticoke Rivers and the north end of Chesapeake Bay in 1999 and from Virginia's Mataponi and Rappahannock Rivers in 2001. A total of 10 microsatellite loci exhibited between two and 27 alleles per locus with observed heterozygosities between 0.255 and 0.893. The 10-locus estimate of R(ST) among the six tributaries was -0.0065 (95% confidence interval -0.0198 to 0.0018). All R(ST) and all but one theta estimates for pairs of populations were not significantly different from zero. Reanalysis of Chesapeake Bay striped bass mtDNA data from two previous studies estimated population differentiation between theta=-0.002 and 0.160, values generally similar to mtDNA population differentiation predicted from microsatellite R(ST) after adjusting for reduced effective population size and uniparental inheritance in organelle genomes. Based on mtDNA differentiation, breeding sex ratios or gene flow may have been slightly male biased in some years. The results reconcile conflicting past studies based on different types of genetic markers, supporting a single Chesapeake Bay management unit encompassing a panmictic striped bass breeding population. PMID- 15829987 TI - Genomics: the chicken genome sequence. PMID- 15829988 TI - PDE-5 inhibitor, Vardenafil, increases circulating progenitor cells in humans. AB - Bone marrow-derived endothelial progenitor cells (EPCs) originate from haematopoietic stem cells in bone marrow and migrate into the peripheral circulation to promote endothelial repair and neovascularization. The number of circulating progenitor cells is reduced in patients with cardiovascular risk factor. The aim of our study was to determine the number of these cells in healthy patients and to evaluate the effect of Vardenfil, a phosphodiesterases-5 (PDE-5) inhibitor, in the number of circulating EPCs. In our study, we found a significant increase in the number of these cells after the drug administration. PMID- 15829989 TI - Do sexual dysfunctions get better during dialysis? Results of a six-month prospective follow-up study from Turkey. AB - Dialysis improves most symptoms of end-stage renal disease (ESRD), yet many patients continue to experience sexual dysfunction (SD) during the dialysis treatment. The aim of this preliminary study was to evaluate the frequency and the course of SD during a 6-month dialysis treatment. Additionally, relationships between the level of depression, cognitive impairment and biochemical parameters of SD were also assessed. The subjects were 43 ESRD (25 male and 18 female) on dialysis treatment for at least 12 months. SD was assessed using the Arizona Sexual Experiences Scale (ASEX); the level of depression and cognitive impairment were assessed using the Hamilton Depression Rating Scale (HDRS) and Mini Mental Status Exam (MMSE). Several biochemical parameters were also assessed. All assessments were carried out at baseline and repeated at 6-month follow-up. Of 43 patients, 20 (47%) and 18 (42%) complained of SD at baseline and at 6-month assessments, respectively. Of 25 males, nine (36%) and seven (28%) patients described SD at baseline and 6-month assessments, respectively; erectile dysfunction was the most frequent complaint. Of 18 females, 11 (61%) and 11 (61%) patients reported SD at baseline and 6-month assessments, respectively; difficulties with arousal and reaching orgasm were the most frequent complaints. Both total and item-by-item comparisons of baseline and 6 months ASEX scores did not reveal any significant changes during 6-month period, indicating that patient's sexual functions do not improve with dialysis treatment. For female patients, HDRS scores were significantly higher in patients with SD at baseline (t = 2.15, P = 0.05) and at 6-month follow-up (t = 2.44, P = 0.03) assessments; after excluding the effects of age and duration of dialysis for females using regression analysis, HDRS still significantly (t = 4.02, P = 0.003) associated with the SD. This preliminary prospective study suggests that SD is frequent in dialysis patients, does not remit with dialysis treatment, associated with depression in female patients, and much clinical attention is indicated. PMID- 15829990 TI - Erectile dysfunction: to prescribe or not to prescribe? AB - Even with the help of modern pharmacology, treatment of erectile dysfunction often remains complex, and requires taking into account the social, psychological, and behavioral dimensions of the disorder. This brief report presents a case study highlighting this complexity. PMID- 15829991 TI - Efficacy of sildenafil as the first-step therapeutic tool for Japanese patients with erectile dysfunction. AB - The aim of this study was to assess the efficacy of sildenafil as the first-step tool for erectile dysfunction (ED) in Japanese males. Between March 1999 and March 2003, 281 patients were prescribed five tablets of sildenafil (50 mg) as the first step in the therapeutic management of ED. Of the 281 patients, 206 were evaluable patients. The overall success rate in achieving sexual intercourse in subjects after taking sildenafil was 77.2% (159/206), while 22.8% (47/206) were unsuccessful. The success rates in men with functional ED and organic ED were 91.4% (85/93) and 65.5% (74/113), respectively (P<0.0001). Overall, transient adverse effects of sildenafil occurred in 16 (8%) males. Intolerable adverse effects (edema and dizziness) occurred in only 1% of patients (2/206). Sildenafil citrate may be recommended as the first choice drug for ED because of its high success rate and low invasiveness. PMID- 15829992 TI - Targeting of VEGF-mediated angiogenesis to rat myocardium using ultrasonic destruction of microbubbles. AB - Myocardial angiogenesis mediated by human vascular endothelial growth factor 165 (hVEGF165) cDNA was promoted in rat myocardium using an in vivo-targeted gene delivery system known as ultrasound-targeted microbubble destruction (UTMD). Microbubbles carrying plasmids encoding hVEGF165, or control solutions were infused intravenously during ultrasonic destruction of the microbubbles within the myocardium. Biochemical and histological assessment of gene expression and angiogenesis were performed 5, 10, and 30 days after UTMD. UTMD-treated myocardium contained hVEGF165 protein and mRNA. The myocardium of UTMD-treated animals showed hypercellular foci associated with hVEGF165 expression and endothelial cell markers. Capillary density in UTMD-treated rats increased 18% at 5 days and 33% at 10 days, returning to control levels at 30 days (P<0.0001). Similarly, arteriolar density increased 22% at 5 days, 86% at 10 days, and 31% at 30 days (P<0.0001). Thus, noninvasive delivery of hVEGF165 to rat myocardium by UTMD resulted in significant increases in myocardial capillary and arteriolar density. PMID- 15829993 TI - Intracellular trafficking of adeno-associated viral vectors. AB - Adeno-associated virus (AAV) has attracted considerable interest as a gene therapy vector over the past decade. In all, 85% of the current 2052 PubMed references on AAV (as of December 2004) have been published in the last 10 years. As researchers have moved forward with using this vector system for gene delivery, an increased appreciation for the complexities of AAV biology has emerged. The biology of recombinant AAV (rAAV) transduction has demonstrated considerable diversity in different cell types and target tissues. This review will summarize the current understanding of events that control rAAV transduction following receptor binding and leading to nuclear uptake. These stages are broadly classified as intracellular trafficking and have been found to be a major rate-limiting step in rAAV transduction for many cell types. Advances in understanding this area of rAAV biology will help to improve the efficacy of this vector system for the treatment of inherited and acquired diseases. PMID- 15829994 TI - Enhanced expression of glutamate decarboxylase 65 improves symptoms of rat parkinsonian models. AB - In this study, we report the amelioration of parkinsonian symptoms in rat Parkinson's disease (PD) models, as a result of the expression of glutamate decarboxylase (GAD) 65 with a modified cytomegalovirus (CMV) promoter. The transfer of the gene for gamma-amino butryic acid (GAD), the rate-limiting enzyme in gama-amino butrylic acid (GABA) production, has been investigated as a means to increase inhibitory synaptic activity. Electrophysiological evidence suggests that the transfer of the GAD65 gene to the subthalamic nucleus (STN) can change the excitatory output of this nucleus to inhibitory output. Our in vitro results also demonstrated higher GAD65 expression in cells transfected with the JDK promoter, as compared to cells transfected with the CMV promoter. Also, a rat PD model in which recombinant adeno-associated virus-2 (rAAV2)-JDK-GAD65 was delivered into the STN exhibited significant behavioral improvements, as compared to the saline-injected group. Interestingly, we observed that these behavioral improvements were more obvious in rat PD models in which rAAV2-JDK-GAD65 was injected into the STN than in rat PD models in which rAAV2-CMV-GAD65 was injected into the STN. Moreover, according to electrophysiological data, the rAAV2-JDK GAD65-injected group exhibited more constant improvements in firing rates than did the rAAV2-CMV-GAD65-injected group. These data indicate that the JDK promoter, when coupled with GAD65 expression, is more effective with regard to parkinsonian symptoms than is the CMV promoter. PMID- 15829995 TI - Ultrasound-mediated delivery of TIMP-3 plasmid DNA into saphenous vein leads to increased lumen size in a porcine interposition graft model. AB - Progressive saphenous vein graft (SVG) narrowing and occlusion remains a major limitation of coronary artery bypass grafting and is an important target for gene therapy. Ex vivo adenoviral gene transfer of tissue inhibitor of metalloproteinase 3 (TIMP-3) reduces adverse SVG remodelling postarterialization, but concerns remain over the use of viral vectors in patients. Ultrasound exposure (USE) in the presence of echocontrast microbubbles (ECM) substantially enhances nonviral gene delivery. We investigated the effects of ultrasound enhanced gene delivery (UEGD) of TIMP-3 plasmid on vascular remodelling in porcine SVG. Maximal luciferase activity (3000-fold versus naked plasmid alone) and TIMP-3 transgene expression in porcine vascular smooth muscle cells in vitro was achieved using USE at 1 MHz, 1.8 mechanical index (MI), 6% duty cycle (DC) in the presence of 50% (v/v) BR14 ECM (Bracco). These conditions were therefore utilized for subsequent studies in vivo. Yorkshire White pigs received carotid interposition SVG that were untransfected or had undergone ex vivo UEGD of lacZ (control) or TIMP-3 plasmids. At 28 d postgrafting, lumen and total vessel area were significantly greater in the TIMP-3 group (10.1+/-1.2 and 25.5+/-2.2 mm2, respectively) compared to untransfected (6.34+/-0.5 and 20.8+/-1.9 mm2) or lacZ transfected (6.1+/-0.7 and 19.7+/-1.2 mm2) controls (P<0.01). These data indicate that nonviral TIMP-3 plasmid delivery by USE achieves significant biological effects in a clinically relevant model of SV grafting, and is the first study to demonstrate the potential for therapeutic UEGD to prevent SVG failure. PMID- 15829996 TI - Economic issues in coronary heart disease prevention in India. PMID- 15829997 TI - Increased circulating concentrations of asymmetric dimethylarginine (ADMA) in white coat hypertension. AB - Elevated plasma levels of the endogenous nitric oxide (NO) synthase inhibitor asymmetric dimethylarginine (ADMA) contribute to endothelial dysfunction and seem to be a predictor for cardiovascular mortality. Elevated ADMA plasma concentrations have been demonstrated in patients with hypertension. However, the plasma concentrations of ADMA in white coat hypertension (WCH) has not been previously studied. The aim of this study was to evaluate ADMA in WCH and compare with normotensive (NT) and hypertensive (HT) patients. We also evaluated the relation between ADMA and NO in these three groups. For this purpose, 34 NT, 34 white coat hypertensive (clinical hypertension and ambulatory daytime blood pressure <135/85 mmHg) and 34 HT patients were recruited in this study. The subjects were matched for age, gender, body mass index (BMI) and the patients with smoking habit, dyslipidaemia and diabetes mellitus were excluded. The ADMA levels were determined by high performance liquid chromatography. Plasma ADMA levels were significantly higher in WCH group than in the NT group (3.21+/-0.49 micromol/l vs 2.84+/-0.58 micromol/l, P=0.046). It was significantly higher in the HT group than in the NTs (4.24+/-0.38 micromol/l, P<0.001). There was also a significant difference between the HT and WCH groups (P<0.001). The WCH subjects had significantly higher levels of NO than the HTs (41.68+/-2.23 vs 32.18+/-2.68 micromol/l; P<0.001) and significantly lower values than the NTs (48.24+/-4.29 micromol/l; P<0.001). In WCH and HT group, there was a negative correlation between ADMA and NO (r=-0.515, P=0.003 and r=-0.389, P=0.034, respectively). In NT subjects, there was no correlation between these two parameters (r=-0.287, P=0.124). The correlation between ADMA and NO was stronger in WCH group than in HT group. Although NO levels in HT patients were lower than WCHs and ADMA levels were higher in HT patients than WCHs, the negative correlation of these two parameters were more pronounced in WCH group. Decreased NO and increased ADMA levels in WCH may indicate endothelial dysfunction. Our data indicate also that WCH represent an intermediate group between NT and HT when endothelial dysfunction is concerned. PMID- 15829998 TI - Normal ambulatory blood pressure: a clinical practice-based comparison of two recently published definitions. AB - The European Society of Hypertension (ESH) has issued guidelines for the detection and treatment of hypertension. According to these guidelines, normal 24 h ambulatory blood pressure (ABP) is defined as lower than 125/80 mmHg. Another publication of ESH recommendations for blood pressure (BP) measurement defines normal awake and asleep blood pressure as lower than 135/85 and 120/70 mmHg, respectively. Our aim was to investigate the compatibility of these two recently proposed ABP cutoffs in clinical practice. We analysed 1495 consecutive ABP measurements. In all, 56% of the subjects were female; age 58 +/- 16 years; body mass index 27 +/- 4 kg/m(2); clinic BP 151+/-22/84 +/- 13 mmHg. Two-thirds were treated for hypertension, and 11% for diabetes. Subjects were classified as having normal 24-h BP if the corresponding value was <125/80 mmHg. Normal awake sleep BP was diagnosed if awake BP was <135/85 mmHg and sleep BP was <120/70 mmHg. Concordance between the cutoffs was found in 93% of the subjects. Among the 7% discordant subjects, 4.5% were hypertensive applying the 24 h, but not awake sleep, BP values, whereas only 2.5% were hypertensive according to awake-sleep, but not 24 h, BP values (P < 0.005). In Conclusion, in real-life ABP measurement, a good agreement was found between two recently issued ABP normality definitions. However, some subjects are classified as hypertensive only according to one of these methods, more often by the 24-h cutoff of 125/80. This discordance may be significant in large-scale clinical BP monitoring. PMID- 15829999 TI - Benidipine improves oxidized LDL-dependent monocyte and endothelial dysfunction in hypertensive patients with type 2 diabetes mellitus. AB - We investigated the effects of long-term benidipine treatment on levels of monocyte and endothelial cell activation markers in hypertensive patients with (n = 28) and without (n = 10) type 2 diabetes mellitus. Benidipine, 4 mg/day, was administered for 6 months; there were no other changes in any of the patients pharmacologic regimens during benidipine treatment. Clinical and biochemical data obtained before and after benidipine administration were compared; all markers were measured by ELISA. The levels of platelet activation markers (CD62P, CD63, and PAC-1), microparticles (monocyte-derived microparticles: MDMPs, and endothelial cell-derived microparticles: EDMPs), chemokines (monocyte chemotactic peptide 1: MCP-1, regulated on activation normally T-cell expressed and secreted: RANTES) and soluble adhesion markers (soluble E-selectin and soluble ICAM-1) differed in the control and hypertension groups. In addition, levels of platelet, monocyte, and endothelial cell activation markers, microparticles, chemokines, and soluble adhesion molecules were higher in hypertensive patients than in those without type 2 diabetes. Furthermore, benidipine administration decreased the concentrations of all these markers. The effect of this drug was significant in diabetes patients with high levels of antioxidized low-density lipoprotein (LDL) antibody. These results suggest that benidipine is effective for the treatment of oxLDL-dependent vascular disorders in hypertensive patients with type 2 diabetes. PMID- 15830000 TI - Appraisal of the knowledge of hypertensive patients and assessment of the role of the pharmacists in the management of hypertension: results of a regional survey. AB - The objectives of the study were to appraise the knowledge which the patients have about their hypertension, and to assess the knowledge and involvement of pharmacists in the management of the hypertensive patient. All the pharmacies in the Poitou-Charentes area were invited both to participate in a training session about arterial hypertension and asked to fill in a pharmacist's questionnaire. Furthermore, each participant was required to submit a patient's questionnaire to 20 consecutive hypertensives. A total of 104 pharmacies and 1015 hypertensive patients participated in the survey. In all, 88% of the patients (n = 893) were aware of their blood pressure (BP) figures, but 68% (349/515) considered themselves wrongly, to be normalized; 39% (n = 350) only had BP figures <140/90 mmHg. They said they had been poorly informed about recommended lifestyle changes. In all, 18% (n = 185) were equipped with an automatic device. A total of 77% (n = 779) were able to give the names of their drugs without the help of the pharmacist. Treatment-related unwanted effects were reported by 8% of the patients (n = 79). Only 18% (n = 29) of the pharmacists were able to provide a correct definition of hypertension. Most of them thought hypertension was well controlled in the general population and considered that both tolerance of and compliance with antihypertensive treatment were satisfactory. They could most often (80%, n = 135) supply a SBPM device, but 58 (36%) only were able to provide relevant advice regarding the recommended procedures. In conclusion, The BP goals and the lifestyle modifications are poorly known by the hypertensives. Pharmacists' knowledge is frequently wrong and should be improved by appropriate training programmes. PMID- 15830001 TI - Enteral administration of a simulated amniotic fluid to very low birth weight neonates. AB - OBJECTIVE: To reduce feeding intolerance among very low birth weight neonates. STUDY DESIGN: A total of 10 neonates with birth weights of 750 to 1250 g were given oral-gastric boluses (2.5 ml/kg every 3 hours) of a solution patterned after amniotic fluid. When milk feedings were begun the milk was mixed with the test solution. The solution was given at a constant daily dose of 20 ml/kg/day while the volumes of milk feedings were gradually increased. When milk feedings reached 80 ml/kg/day the test solution was discontinued. A comparison group consisted of neonates who met study criteria but were cared for during the period immediately preceding the study. The outcome was the number of calories taken enterally over the first 21 days of life. RESULTS: The test solution was begun an average of 27 hours after birth (range, 4 to 45). In the test group the first milk feedings were introduced 74 hours after birth (range, 18 to 144), which was similar to the time milk was introduced in the comparison subjects (79 hours; range, 18 to 168). After milk feedings were started, the test patients had a total of four NPO days (0.4 NPO days per patient) during their first 21 days, while the comparison group had 34 NPO days (3.4 NPO days per patient). During the first 14 days of life the test solution recipients had a median of 26.5 enteral cal/kg/day (range, 4.3 to 68.9), while the comparison neonates had 8.5 (range, 0.2 to 25; p < 0.05). During the first 21 days of life the test solution recipients had a median of 56.9 enteral cal/kg/day (range, 11.5 to 89.4), while the comparison neonates had 19.2 (range, 0.9 to 52.8; p < 0.05). CONCLUSION: In all, 10 VLBW infants tolerated the test solution for periods up to 14 days with no significant adverse effects. A randomized trial to determine whether this solution reduces feeding intolerance among VLBW neonates should be conducted. PMID- 15830002 TI - Vernix caseosa in neonatal adaptation. AB - OBJECTIVES: To characterize vernix caseosa in newborn infants with respect to factors that influence vernix distribution on the skin surface, vernix effects on thermal stability, skin hydration, acid mantle development, and vernix antioxidant properties. STUDY DESIGN: Vernix distribution was determined for 430 infants. Thermal stability was assessed in parallel groups following vernix retention (n=66) and removal (n=64). The effects of vernix retention on skin hydration, pH, erythema, and dryness/scaling were determined. Samples were analyzed for vitamin E before and after UV exposure. RESULTS: Vernix distribution depended upon gestational age, delivery mode, gender, race, and meconium exposure. Retention had no effect on axillary temperatures. Skin hydration was significantly higher for vernix-retained skin. Skin pH and erythema were significantly lower with retention. Vitamin E levels were decreased by ultraviolet radiation. CONCLUSIONS: Vernix is a naturally occurring barrier cream with multiple salubrious effects, which support its retention on the skin surface at birth. PMID- 15830003 TI - Very low prevalence of endophthalmitis in very low birthweight infants who survive candidemia. AB - OBJECTIVE: Candida species often cause sepsis in very low birthweight (VLBW) infants, leading to formal ophthalmologic evaluation for endophthalmitis. Our experience suggests that endophthalmitis is rare in this setting, and retinal vascular compromise in extreme prematurity may decrease the risk. We studied the prevalence of endophthalmitis in surviving VLBW infants with candidemia. STUDY DESIGN: Epidemiologic data and presence of ROP and endophthalmitis were ascertained for all VLBW infants with candidemia at our institution from 1994 to 2001. RESULTS: A total of 123 infants were included. Median EGA was 25 weeks (range, 23 to 32) and median birthweight was 735 g (range, 426 to 1460). Of these 123, only one had transient retinal findings (prevalence 0.8%; 95% confidence interval 0 to 4%), which resolved during therapy. In no case was either the duration of therapy or the outcome of candidemia altered by retinal examination. CONCLUSIONS: Aggressive treatment of candidemia has made endogenous endophthalmitis rare in candidemic VLBW infants. PMID- 15830004 TI - Pulmonary hypertension and right ventricular dysfunction in growth-restricted, extremely low birth weight neonates. AB - This case series reports an acute episode of hypoxemia and systemic hypotension in seven infants under 1 kg, taking place several hours or days after birth, after a period of stability and in the absence of significant lung disease. These patients were growth-restricted at birth and had a history of chronic fetal hypoxia and oligohydramnios. Pulmonary hypertension and right ventricular dysfunction were found by echocadiography. Right ventricular ejection fraction was significantly depressed during the acute episode, compared to baseline values measured after recovery. The timing of symptoms seemed related to ductus arteriosus closure or constriction. Oxygenation and right ventricular function improvement occurred within a few days under ventilatory and inotropic support, while milrinone was administered in five cases. In conclusion, pulmonary hypertension is a rare but significant cause of hypoxemia in preterm infants, and pulmonary vasodilator therapy should be considered in the presence of right ventricular dysfunction. PMID- 15830005 TI - Procalcitonin as a screening test for late-onset sepsis in preterm very low birth weight infants. AB - OBJECTIVE: To compare the utility of procalcitonin (PCT) vs C-reactive protein (CRP) as indicators of late-onset neonatal sepsis in very low birth weight (VLBW) infants. METHODS: PCT and CRP levels were measured in VLBW infants with suspected sepsis and controls. Comparisons were made between infected vs noninfected infants. Using cutoff values of 0.5 and 1.0 ng/ml for PCT and 0.8 mg/dl for CRP, sensitivity, specificity, positive and negative predictive values were calculated to evaluate these assays as potential predictors of late-onset sepsis. RESULTS: A total of 67 infants were evaluated. Mean PCT levels were significantly higher in the infected group (5.41 ng/ml) compared to the noninfected group (0.43 ng/ml) (p < 0.001). At a cut off value of 0.5 ng/ml, the sensitivity of PCT was 97%, whereas that of CRP was 73% in predicting late-onset sepsis. At a PCT cutoff of 1.0 ng/ml, sensitivities of PCT and CRP were similar (72% each). CONCLUSION: PCT (0.5 ng/ml) is more sensitive than CRP in predicting late-onset sepsis in VLBW infants. PMID- 15830006 TI - CD40 activation of B-CLL cells is associated with augmented intracellular levels of CD79b and increased BCR expression in a subset of patients. PMID- 15830007 TI - Postremission therapy for children with acute myeloid leukemia: the children's cancer group experience in the transplant era. AB - We reviewed consolidation therapy results and analyzed postremission outcomes for 1464 children less than 21 years old at diagnosis in five consecutive Children's Cancer Group acute myeloid leukemia trials between 1979 and 1996. Children in remission were allocated to allogeneic bone marrow transplantation (BMT) (N=373) in first remission, if a matched family donor was available. Remaining children were assigned consolidation chemotherapy (N=688) or autologous purged BMT (N=217), or withdrew from study before assignment, or with unknown data (N=186). Overall and disease-free survival were superior for children assigned allogeneic transplants. High (>50,000/microl) diagnostic white blood cell (WBC) count was prognostic for inferior outcome, but French-American-British (FAB) subtypes were not. Inv(16) is a favorable karyotypic feature for children in first remission and t(8;21) is not. Allogeneic transplantation benefit was evident in most children, including those with high or low diagnostic WBC count, each FAB subtype, and t(8;21), but was not seen in children with inv(16). Therefore, these data suggest reserving matched related donor allogeneic transplantation for children with inv(16) for second remission, but not those with t(8;21). PMID- 15830008 TI - High serum bone-specific alkaline phosphatase level after bortezomib-combined therapy in refractory multiple myeloma: possible role of bortezomib on osteoblast differentiation. PMID- 15830009 TI - Gender aspects in chronic myeloid leukemia: long-term results from randomized studies. AB - Gender-related aspects in chronic myeloid leukemia (CML) have not been studied well. We therefore analyzed 856 patients with Ph/BCR-ABL-positive CML from the German randomized CML-studies I (interferon alpha (IFN) vs hydroxyurea (HU) vs busulfan) and II (IFN+HU vs HU alone). The median observation time was 8.6 years. A total of 503 patients (59%) were male. Female patients were older (51 vs 46 years; P<0.0001), presented with lower hemoglobin (11.7 vs 12.5 g/dl; P<0.0001), higher platelet counts (459 vs 355 x 10(9)/l; P<0.0001), smaller spleen size (3 vs 4 cm below costal margin; P=0.0097), a lower rate of additional cytogenetic aberrations (9 vs 15%; P=0.018) and a less favorable risk profile (P=0.036). The transplantation rate was 14% for female (n=48) and 22% for male patients (n=113). Median survival was longer in female patients (58 vs 49 months; P=0.035) mainly attributable to better survival in the low- and intermediate-risk groups and, independent from risk groups, in the HU group. These results were confirmed by matched-pair analyses based on German population data (n=496, 59 vs 45 months; P=0.0006). This is the first analysis of gender aspects in CML using randomized trials. It demonstrates the relevance of analyses of gender differences in CML and in malignant disease at large. PMID- 15830010 TI - Mobilisation with G-CSF in healthy donors promotes a high but temporal deregulation of genes. PMID- 15830011 TI - Reduced-intensity conditioning lowers treatment-related mortality of allogeneic stem cell transplantation for chronic lymphocytic leukemia: a population-matched analysis. AB - To elucidate whether reduced-intensity conditioning (RIC) decreases treatment related mortality (TRM) after allogeneic stem cell transplantation (allo-SCT) for chronic lymphocytic leukemia (CLL), we retrospectively compared 73 RIC cases from a recent EBMT survey with 82 patients from the EBMT database who had undergone standard myeloablative conditioning (MC) for CLL during the same time period. The two populations were matched by adjusting the primary risk factor, the conditioning regimen, in a series of Cox models for age, sex, donor type, remission status at transplant and analyzed for its effect on TRM, relapse incidence, event-free (EFS) and overall survival (OS). After adjustment, a significant reduction of TRM became evident for the RIC population (hazard ratio (HR) 0.4 (95% confidence interval 0.18-0.9); P=0.03). On the other hand, RIC was associated with an increased relapse incidence (HR 2.65 (0.98-7.12); P=0.054). There was no significant difference between RIC and MC in terms of EFS (HR 0.69 (0.38-1.25); P=0.22) and OS (HR 0.65 (0.33-1.28); P=0.21). We conclude that RIC appears to favorably influence TRM after allo-SCT for CLL. This observation, as well as possible detrimental effects of RIC on relapse risk, should be confirmed by prospective studies. PMID- 15830012 TI - Correlation of CD33 with poorer prognosis in childhood ALL implicates a potential of anti-CD33 frontline therapy. PMID- 15830013 TI - Expression of activation antigens on lymphocyte surface and circulating platelet leukocyte aggregates in ischaemic heart disease. AB - BACKGROUND: Data from literature documented the role of the activation of circulating T lymphocytes and increased leukocyte adhesion to blood platelets in the destabilisation of an atheromatous plaque and progression of ischaemic heart disease to acute coronary syndrome. AIM: To assess whether there is an increased proportion of activated T lymphocytes and platelet-leukocyte aggregates (PLA) in the circulating blood in patients with myocardial infarction or with stable angina, and to examine whether these changes are related to the progression of clinical symptoms or coronary angiography results. METHODS: The study group consisted of 36 patients with ST-segment elevation acute myocardial infarction (STEMI), confirmed by elevated troponin T level (36 patients, 26 males, 10 females, mean age 61.8 years, range 42-78 years), 30 patients with stable angina and single-vessel disease (24 males, 6 females, mean age 58.8 years, range 43-69 years), and 20 control healthy age and gender-matched subjects. Lymphocyte activation was evaluated by the assessment of T lymphocytes CD3+ /CD69+, CD3+/HLA DR+ and CD4+/CD154+. The PLA assessment, including platelet-granulocyte aggregates (PGA), platelet-monocyte aggregates (PMA) and platelet-lymphocyte aggregates (PlymphA) was based on the measurement of the proportion of CD45+/CD41a+ cells with the use of flow cytometry. RESULTS: The proportion of T lymphocytes CD3+/HLA-DR+, CD3+/CD69+ and CD4+/CD154 was significantly higher in patients with STEMI than in controls, and T lymphocytes CD3+/CD69+ - significantly higher in STEMI group than in both patients with angina or controls. There was no significant relationship between the proportion of activated lymphocytes and duration of anginal pain, troponin T concentration or the number of coronary vessels with critical stenosis. The proportion of PGA, PMA and PlymphA was significantly greater in STEMI patients than in patients with angina or controls. There was a significant positive correlation between the proportion of PLA and PMA, and the duration of anginal pain. Patients with stable angina had a significantly higher proportion of T lymphocytes CD3+/HLA-DR+ compared with controls. CONCLUSIONS: Circulating T lymphocyte activation is present in ischaemic heart disease. This phenomenon is more pronounced in patients with acute MI than in those with stable angina, and is not related to the degree of cardiac injury. An increased formation of platelet-leukocyte aggregates is also present in patients with acute MI. PMID- 15830014 TI - Atrial natriuretic peptide for monitoring cardiovascular system performance following pacemaker implantation. AB - BACKGROUND: The effects of different modes of pacing on cardiac performance have been investigated in a few small studies, and the results are not conclusive. AIM: To assess the effects of different modes of pacing on atrial natriuretic peptide (ANP) plasma concentration and left ventricular ejection fraction (LVEF). METHODS: The study group consisted of 55 subjects without symptomatic CHF. Forty patients were in sinus rhythm and had a-v block. Twenty of them, aged 71-90 years, mean 77.5+/-5.9 years, received VVI pacemaker, and the remaining 20 patients aged 49-81 years, mean 68.9+/-11.9 years, received DDD or VDD pacemakers. The control group consisted of 15 healthy volunteers aged 58-80 years, mean 72.6+/-2.8 years. Blood samples for alpha ANP concentration were taken before pacemaker implantation and 7 as well as 30 days after pacemaker implantation. In controls, a single alpha ANP measurement was performed. Transthoracic echocardiography was performed in all patients and controls at the same time-points as ANP assessment in order to calculate LVEF. RESULTS: Before pacemaker implantation, ANP plasma concentration was the highest in VVI patients and was significantly greater in patients than in controls. Following VVI pacemaker implantation, ANP level significantly decreased, both 7 and 30 days after implantation when compared with baseline values. Also following implantation of dual-chamber pacing system, ANP concentration significantly decreased. The baseline LVEF values were significantly higher in patients than in controls. In patients with VVI pacemakers, LVEF significantly increased, both 7 and 30 days after implantation. In patients with dual-chamber pacemakers, LVEF tended to increase 7 days after implantation and slightly decreased 30 days after implantation, however, the differences were not significant. CONCLUSIONS: ANP level decreases following pacemaker implantation in patients with advanced a-v block which suggests an improvement of cardiac performance. Assessment of ANP may be useful in selection of pacing mode. PMID- 15830015 TI - Prevalence of arterial hypertension in children aged 9-14 years, residents of the city of Lodz. AB - BACKGROUND: Hypertension is one of the most common disorders in adults. Although it may be already present in children and adolescents, it is usually undetected. There is a lack of data on the prevalence of hypertension in this age-group in Poland. AIM: To assess the prevalence of hypertension in children and adolescents, residents of the city of Lodz. METHODS: Children and adolescents aged 9-14 years, attending one primary and one secondary school, were examined. Blood pressure (BP) was measured three times with the use of a mercury sphygmomanometer. Systolic BP was determined by the first Korotkoff sound, and diastolic BP - by the fifth Korotkoff sound. Hypertension was diagnosed when the value of systolic and/or diastolic BP exceeded the 95th percentile of normal values, taking into account height, gender and age. RESULTS: In total, 637 children and adolescents (349 girls and 288 boys) were examined. The first BP measurements revealed abnormal values in 13.2% of examined subjects, slightly more often in girls than in boys (14.6% vs 11.4%, NS). Triple BP measurements revealed the presence of hypertension in 3.3% of examined subjects (2.9% of girls and 3.8% of boys, NS). Hypertension was diagnosed by triple measurement in a significantly lower proportion of subjects than by a single screening examination (p<0.05). CONCLUSIONS: Hypertension is present in a significant proportion of children and adolescents who live in the city of Lodz, which shows the need for a systematic screening. A proper detection of hypertension requires repeated BP measurements. PMID- 15830016 TI - What primary care physicians know about diagnosis and treatment of chronic heart failure. AB - BACKGROUND: The majority of patients with chronic heart failure (CHF) are treated by primary care physicians. Recent studies have shown that their knowledge about diagnosis and treatment of CHF is not satisfactory. AIM: To evaluate the knowledge of primary care physicians on the diagnosis and treatment of patients with CHF. METHODS: The survey consisting of 23 questions on the diagnosis and treatment of CHF, was filled in by 116 primary care physicians, practising in the city of Poznan. The questions dealt with the most typical characteristics of CHF, including history, physical examination, diagnosis of diastolic CHF and medical treatment, according to the recent guidelines of the European Society of Cardiology. RESULTS: More than 90% of physicians regarded history and physical examination as the most important elements required for establishing the diagnosis of CHF. Ankle oedema, dyspnea and fatigue were listed as the most typical CHF symptoms. Almost 80% of surveyed physicians routinely performed ECG and chest radiogram in order to diagnose CHF. Echocardiography was used by only 20% of physicians, and the term "diastolic CHF" was familiar to only one third of them. Angiotensin converting enzyme inhibitors (ACEI) were used by more than two thirds of doctors, and beta-blockers--by one third. The correct initial recommended dose of enalapril was known to more than half of the physicians, and the target dose--to only one fourth of them. The correct initial recommended dose of carvedilol was known to almost half of the physicians, and the target dose--to only 3% of them. Diuretics were prescribed by 62% of physicians, spironolactone- by half of them, and digitalis--by one third of them. CONCLUSIONS: Primary care physicians practising in the Poznan area diagnose CHF mainly on the basis of history, physical examination, ECG and chest radiogram. Echocardiography is underused which may hamper the accuracy of CHF diagnosis. ACEI and beta-blockers are also underused. These findings demonstrate that knowledge about diagnosis and treatment of CHF is insufficient among primary care physicians and indicate the need for educational and systematic changes, including a wider access to echocardiography. PMID- 15830017 TI - Treatment options for post-catheterisation femoral pseudoaneurysm closure. AB - BACKGROUND: Femoral artery pseudoaneurysm (FAP) complicates from 1% to 9% of all coronary angiography procedures and contributes to extended hospitalisation as well as patient discomfort. AIM: To compare three main methods of FAP closure which are used nowadays. METHODS: Seventy-five subjects (38 females, 37 males, mean age 60.8+/-10.4 years) with post-catheterisation FAP were studied. The results of three methods of FAP closure--surgical, local compression and thrombin injection--were compared. RESULTS: Between September 2000 and July 2001, fourteen patients developed FAP; in 9 (64%) patients FAP was closed with repeated prolonged compression whereas the remaining 5 (36%) patients required surgical closure of compression-resistant FAP. We observed that FAPs with longer neck (>10 mm) and primary signs of partial spontaneous coagulation were more prone to self closure as compared to FAPs with short neck and no signs of perimural coagulation (p=0.01). Since July 2001, we introduced ultrasound-guided thrombin injection into FAP sack. The protocol included attempt of closing FAP with probe compression and compression dressing put overnight, and, if unsuccessful, followed by a quick injection of 2 ml of thrombin solution (400-3200 U), guided by ultrasound. During this period, we identified 61 patients with FAP. Out of this group, 5 (8.2%) subjects were referred for surgery without any attempt of thrombin-injection, in 16 (26.2%) patients FAP was closed with probe compression and dressing put overnight, and in the remaining 40 (65.6%) subjects ultrasound guided thrombin-injection was performed. Thrombin injection into FAP sack caused closure of its cavity and neck in all patients, however, five patients required additional thrombin injection during the same session, and 2 (5.0%) patients- during the next procedure. No peri-procedural complications were observed. The duration of hospital stay shortened from a mean of 26.6+/-14.5 days in surgically treated patients to 7.9+/-6.7 in those in whom FAPs were closed with compression, and to 4.6+/-2.6 days in those treated with thrombin (p<0.001). During a mean follow-up of 11+/-8.1 months, we re-examined 32 (80.0%) patients in whom FAP was closed with thrombin injection. No long-term thrombotic or embolic complications were observed. However, in 2 (6.3%) patients FAP cavity did not undergo complete resorption after 6 and 12 months of follow-up. CONCLUSIONS: Thrombin-induced closure of femoral pseudoaneurysm is a quick, safe and effective method, shortening hospitalisation time. In our Department this procedure replaced the prolonged and painful compression method. PMID- 15830018 TI - [Negative delta wave mimicking old inferior myocardial infarction--a case report]. AB - A case of a 64-year-old male diagnosed with remote inferior myocardial infarction (MI), is presented. Electrophysiological examination revealed the presence of an accessory pathway with a negative delta wave in inferior leads mimicking pathological Q wave. Differential diagnosis of pathological Q waves is discussed. PMID- 15830019 TI - [Variant angina mimicking right ventricular infarction--a case report]. AB - A case of a 59-year-old man with recurrent angina associated with ST-segment elevation in leads II, III, aVF and V4R-V6R, cardiogenic shock and complete atrio ventricular block, is presented. Coronary angiography was normal. Therapy with calcium channel blockers was effective. Difficulties in the treatment of patients with variant angina are discussed. PMID- 15830020 TI - [Non-surgical retrieval of coronary stent from the left ventricle following failed stent deployment in the left anterior descending coronary artery--a case report]. AB - A case of a 66-year-old male with acute myocardial infarction (MI) treated with streptokinase is presented. Due to the recurrence of angina six days from the acute phase of MI, the patient underwent coronary angiography which revealed critical stenosis of the left anterior descending coronary artery. During intracoronary intervention, stent was lost and migrated to the left ventricular cavity, being trapped in chordae tendinae of the posterior mitral leaflet. The procedure was stopped. After transferring the patient to our department a few days later, a successful non-surgical retrieval of stent with the use of biopsy forceps was performed. Treatment of dislodged coronary stents is discussed. PMID- 15830021 TI - [Pharmacological treatment of chronic heart failure in patients with renal failure]. PMID- 15830022 TI - [10th anniversary of "Klub 30"]. PMID- 15830023 TI - [10th anniversary of "Klub 30"]. PMID- 15830024 TI - ["Klub 30"]. PMID- 15830026 TI - [Echocardiographic terminology in Polish. Recommendations of Polish Cardiac Society]. PMID- 15830027 TI - [Electrocardiographic signs of anterior myocardial infarction caused by the occlusion of the right ventricular branch of the right coronary artery--a case report]. AB - Elektrocardiographic signs of anterior myocardial infarction caused by the occlusion of the right ventricular branch. A case of a 72-year-old male with electrocardiographic symptoms of anterior myocardial infarction resulting from the right ventricular branch occlusion is presented. The mechanisms of eliscrepancy between angiographic and electrocardiographic findings are discussed and diagnostic as well as therapeutic procedures are described. PMID- 15830028 TI - [Cor triatriatum diagnosed in a patient with severe mitral insufficiency]. PMID- 15830029 TI - [Angiogram of the month]. PMID- 15830030 TI - [Long-term effects of radiofrequency ablation of ventricular tachycardia originating from the aortic cusps]. PMID- 15830031 TI - [How to diagnose ischaemic heart disease in females?]. PMID- 15830032 TI - The role of PPARgamma-dependent pathway in the development of cardiac hypertrophy. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors belonging to the nuclear receptor superfamily. PPARs have three isoforms, alpha, beta (or delta) and gamma. It has been conceived that PPARgamma is expressed predominantly in adipose tissue and promotes adipocyte differentiation and glucose homeostasis. Recently, synthetic antidiabetic thiazolidinediones and natural prostaglandin D2 (PGD2) metabolite, 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2), have been identified as ligands for PPARgamma. Following demonstration that PPARgamma is present in a variety of cell types, further study of PPARgamma has been conducted. Although activation of PPARgamma appears to have beneficial effects on atherosclerosis and heart failure, it is still largely uncertain whether PPARgamma ligands prevent the development of cardiovascular diseases. Recent evidence suggests that some benefit from the antidiabetic agents known as thiazolidinediones may occur through PPARgamma-independent mechanisms. In this review, we report on the latest developments concerning the study of PPARs and summarize the roles of the PPARgamma-dependent pathway in cardiovascular diseases. PMID- 15830033 TI - The impact of vascular risk factors on erectile function. AB - The main cause of erectile dysfunction is organic in nature, with vascular etiologies being the most common risk factors. The incidence of erectile dysfunction increases with the number of vascular comorbidities such as hypertension, diabetes, hyperlipidemia, smoking and atherosclerosis. The earliest signal of endothelial damage in men with vascular risk factors is the manifestation of erectile dysfunction. The penis is a barometer of the body's endothelial function, so it is reasonable then to correlate vascular pathologies as direct causes of erectile dysfunction. Moreover, erectile dysfunction may be the first clinical presentation of any of these comorbidities, with the vascular endothelium playing a pivotal role in regulating vascular homeostasis of the corpora cavernosa. This article addresses the impact of vascular risk factors on erectile function based on current evidence. PMID- 15830034 TI - Renal and humoral pathophysiological actions of angiotensin II in congestive heart failure. AB - The renin-angiotensin-aldosterone axis is an integral component linking the renal humoral system to the cardiovascular system. It is involved in the normal control of blood pressure and intravascular volume. Its activity is also enhanced in pathologic states, namely congestive heart failure, in which stimulation of the axis leads to further deleterious effects on the heart. The well-established dogma that the renin-angiotensin-aldosterone system (RAAS) is a linear cascade is evolving into a vision of this system as a more complex process. It is now known that angiotensin has several subtypes. Each subtype is a ligand at several receptor subtypes and these interactions are not mutually exclusive. The aim of this review is to discuss the different angiotensin subtypes, their receptor interactions and their pathophysiological roles in humoral and renal functions in congestive heart failure. In addition, we will also review the different therapeutic approaches that interrupt the RAAS and the evidence that supports their utility in congestive heart failure (CHF). PMID- 15830035 TI - Insights into cardiovascular therapy: a report from the 2005 Annual Meeting of the American College of Cardiology. PMID- 15830037 TI - [Not Available]. PMID- 15830038 TI - [History of obstetrics in nineteenth-century Brazil]. AB - This research aimed at recovering the peculiarities of the development of Obstetrical Art in Brazil during XIX century, starting in 1808, when a legal act signed by D. Joao VI included this discipline in the initial curriculum of the schools of medicine and surgery founded in Bahia and Rio de Janeiro. Research was undertaken through the review of 83 medical theses in the field of Obstetrics, produced in Bahia and Rio de Janeiro in XIX century. The data showed that Obstetrical Art was traditionally performed by women called "aparadeiras" (catchers) or "comadres", who cared for women before, during and after delivery, and provided assistance in other situations, like venereal diseases and abortion. The inclusion of physicians-obstetricians in this practice led not only to a scrutinizing of the female body, but also to the production of an anatomical and physiological knowledge of the female organism from a male standpoint. PMID- 15830039 TI - [Cesarean section in Brazil: incidence, trends, causes, consequences and suggestions for change]. AB - The authors analyse the current cesarean section situation in Brazil. In the last decade it has reached extremely high levels of incidence, higher than any other country. Social-economic and regional differences are established through the available national data on the cesarean section incidence. There is a higher incidence on wealthier regions and women. The factors influencing this high incidence, including socio-cultural, obstetric care organization, legal and institutional ones are also considered. Special attention is given to the problem of female surgical sterilization which is forbidden in the country and is performed through inadequate indication of a cesarean section. Consequences for maternal and perinatal morbidity and mortality, the effects on population fertility and extra public expenses are considered. Interventions and attitude changes are proposed at several levels to reverse this increasing tendency of cesarean section incidence in the country. PMID- 15830040 TI - [Female mortality in Brazil: the weaker sex or stronger sex?]. AB - Population ageing in Brazil has been more rapid and more intense among women. This phenomenon is well described in developed countries where mortality rates are higher for men than women. In this regard, the analysis of mortality patterns by cause contributes to elucidate the determinant factors of the present situation in Brazil and provides indications of some future trends in female mortality. This is especially important due to the fact that in Brazil the social role of women has experienced great changes. This study presents data on mortality from ten capital cities in 1985, showing age-standardized overall and cause-specific mortality rates for five of the main groups of causes by sex. Ratios and differences effect estimators were used. The results revealed that regional patterns are associated with the urban and industrial processes with greater differences by sex in more developed regions. External causes and cardiovascular diseases are the main factors responsible for higher mortality among men with special emphasis on violent deaths. It is inferred that the present trend will be maintained, though it is possible that mortality differences by sex could decrease in the near future. The authors discuss that longer survival among women in Brazil does not reveal better life conditions. PMID- 15830041 TI - [We control our bodies: the biological and social dialectic]. AB - This article aims at reviewing the discussion of biological and social factors in the analysis of women's social condition. With the appearance of a feminist perspective, the dominance of earlier biologically-based explanations was substituted by an emphasis on the social construction of female identity. Even when women's identification with the body and with nature, and their secondary status, were considered universal, biological determinism was rejected. In this process of re-definition of the object of study, the ideological role of science was pointed out, since male dominance in science and society accompanied the historical tendency which relegated "the woman question" to the sphere of natural fact. Although growing awareness of the socially-constructed nature of scientific activity itself is producing a tendency to abandon the biological/social dichotomy at the conceptual level, differences between men and women in the reproductive sphere continue to exist. It is argued that analysis of reproduction requires characterization of the sexes as biosocial entities in relationship, situated in specific historical contexts, and that in modern society women are subject to a double reproductive contradiction. PMID- 15830042 TI - [Framework for the study of the relationships between work, women and health]. AB - The present paper aims at showing the importance of the gender approach in the theoretical construction of the field of worker's health since men and women are exposed to different work conditions within the productive process. In the case of female workers, we point to the liaison of capitalism and patriarchy as determinants of women's oppression in the hierarchical relationships of labor as well as to the hidden social responsibility of the housework, which is carried out in the private sphere thus keeping women distant from the social and political world and generating strong impacts on their health. According to the sexual division of labor, female work in industry is restricted to rather repetitive tasks that require a great nervous resistance - not especially healthy conditions that lead to a specific consuming pattern. We concluded that changes should occur in the social life schemes, besides breaking with the myths that support the sexual division of labor, so that the aggressive conditions imposed on men and women's health in the work process can be altered. PMID- 15830043 TI - [Is male violence directed towards Eve or Mary?]. AB - This paper analyses the different categories of violent death and other injuries occurred in Bahia. In Brazil, traffic accidents, homicides and suicides correspond to the third most common cause of death. Besides, violent acts tend to increase the emergency health services demand. The data were collected from the written press covering all the homicides, grave injuries and notified rapes during 1989. Police violence appeared as responsible for the death of a high proportion (24%) of young males, followed by the disagreement between criminals (18.6%). In relation to females, the quantitative dimension of crimes are lower, more diffused and registered in the family environment. The data point to the importance of male domination in the interpretation of those domestic violent acts. Resisting the ideology of women's inferiority means avoiding both injuries and homicides - a complex task for Brazilian society. PMID- 15830044 TI - [AIDS in adult women in the Municipality of Rio de Janeiro]. AB - This paper presents the main epidemiological characteristics of AIDS cases observed among adult women in Rio de Janeiro Municipality (Rio) during 1980-89. Higher incidence rates as compared to Brazilian and Canadian women, rising participation of either sexual transmission and EV drug addiction and an increasing involvement of young women were also observed. The disease does not present a random spatial distribution in Rio, and case clusters in definite high risks areas were shown. Tuberculosis was found to be the second most frequent opportunistic infection detected among cases. Finally, some of the main trends and questions related to AIDS control among women in the studied area are discussed. PMID- 15830045 TI - [Induced abortion among low income women: dimensions of the problem]. AB - This paper examines the practice of abortion, especially induced abortion among low income women. The discussion is based on survey data collected between 1984 and 1985 in seven slum communities (favelas) situated in the metropolitan area of Rio de Janeiro, Brazil. Despite restrictive law, induced abortion is extremely frequent. Among married women 21.4 per cent reported experience of induced abortion. Most abortions were performed by physicians, however the quality of care of these procedures can be questioned since almost all induced abortions are illegal there is no possible supervision by health authorities. The incidence of post-abortion complication is very high, especially for those performed by traditional midwifes or by the woman herself. More than 60 per cent of the women were not using contraception at the time of pregnancy. About, 21 per cent reported that they were using the pill. Such a high pill failure rate is inacceptable, and probably was related to incorrect use. This points to the need for a better access to family planning care within the health services. The consequences of the restrictive abortion laws in Brazil are also discussed. Restrictions that in practice prove to have little impact on the practice of induced abortion, appear to be very effective in brooding even more the social economic inequalities. PMID- 15830047 TI - [Propsitions for a relationship between health professionals and women]. PMID- 15830046 TI - [Group consultation: a space for women]. AB - This experience took place in a Medical Center, in Rio de Janeiro, in 1988. The group responsible for it included a social worker, a nutritionist, a nurse and a gynecologist. The aims were: to offer better health care; to discuss the importance, the results and the appropriate therapies of the Pap smear; to offer women an opportunity to meet and discuss their problems, experiences, desires, expectations; and to offer to the health professionals the opportunity to re think their own practices. The methodology used was characterized by making appointments for a medical exam subsequent to a Pap smear; evaluation of each Pap smear result and its appropriate treatment, previous to the group consultation; group discussion about the Pap smear results; and about themes brought up by women clients. The main results were: an increase of 42.7% in the number of women who went for their first medical exam; an increase of ten minutes in the length of each medical exam; better screening for syphilis and gonorrhea, better quality of health care within the perspective of women as integral beings (greater appreciation of a woman s own experiences, be it in terms of her complaints and problems be it in her evaluation of the medical care system). This kind of consultation is of rather recent date and if it becomes more wide-spread it could result in an increase in the number and quality of the medical exam. PMID- 15830056 TI - Is MTHFR polymorphism a risk factor for Alzheimer's disease like APOE? AB - BACKGROUND: The role of methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms as risk factors for the occurrence of Alzheimer's disease (AD) is still controversial. OBJECTIVE: To verify the association between MTHFR and apolipoprotein E (APOE) polymorphisms and Alzheimer's disease. METHOD: This work was conducted as a case-control study. Cases included thirty patients with probable AD. Controls were constituted by 29 individuals without dementia according to neuropsychological tests paired to age, sex, race and educational level. DNA was isolated from peripheral leukocytes of anticoagulated venous blood. Genotyping of APOE and MTHFR were performed by DNA amplification and digestion. The frequences of APOE and MTHFR genotypes were submitted by chi square test corrected by Fisher test; the APOE genotypes, to chi-square linear tendency test and the frequences of MTHFR mutant and AD, by stratificated analysis adjust by Mantel-Haenszel method. RESULTS: There was significant difference about APOE4 and APOE2 in the groups. (p=0.002) The odds ratio increased exponentially with the increased number of E4 allele (chi2 linear tendency test). No significant difference was detected on MTHFR genotypes in both case and control groups. CONCLUSION: The APOE4 is a risk factor and demonstrated a dose-dependent effect while APOE2 allele conferred a protection to AD. The MTHFR mutation had no correlation with AD. PMID- 15830057 TI - Tumor necrosis factor-alpha, interleukin-1beta and interleukin-6 in the cerebrospinal fluid of newborn with meningitis. AB - OBJECTIVE: To analyze the usefulness of determining the cerebrospinal fluid (CSF) levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) for the early diagnosis and evaluation of the prognosis of neonatal meningitis. METHOD: We studied 54 newborn that underwent lumbar puncture. Thirty patients had meningitis and 24 were the control group. CSF and sera were obtained at the moment of suspicion of meningitis and stored at -70 degrees C. Cytokines were performed by enzyme-linked immunosorbent assay method. RESULTS: CSF cytokines were detected in all the newborn with meningitis. TNF alpha was detected in the CSF in 63.3% of the neonates, IL-1beta in 73.3% and IL 6 in 96.6%. The CSF levels were significantly higher than serum in neonates with meningitis. There was no correlation between the CSF levels of cytokines and neurologic complications. CONCLUSION: The detection of TNF-alpha, IL-1beta and IL 6 in the CSF is of great value in order to achieve a early diagnosis of neonatal meningitis. Among the three cytokines analyzed, IL-6 was the best indicator of meningeal inflammation. PMID- 15830058 TI - Prognostic value of non-reactive burst suppression EEG pattern associated to early neonatal seizures. AB - Seizures are the most frequent neurological event in newborns and clinical data suggest that etiology is the dominant factor in long term outcome. However, there are consistent background EEG abnormalities associated to neonatal seizures that are usually related to unfavorable outcome as the burst-suppression pattern. OBJECTIVE: The objective of this study was to correlate clinical and EEG features associated to long-term outcome of newborns with non-reactive burst-suppression (BS) EEG. METHOD: Newborns included in the study were selected from our database and had conceptional age (at the time of first EEG) >37 weeks, EEG recordings with non-reactive BS available for review and clinical follow up. RESULTS: 12 newborns met inclusion criteria, 50% had seizures in the first day of life. Seizures became refractory to treatment in all of them. In 50% the etiology of seizures was considered cryptogenic, 33% had inborn errors of metabolism and 17% had clinical history and neuroimage suggestive of hypoxic-ischemic encephalopathy. The follow-up showed that 7/12 infants deceased, 3 during the first year of life, and one in the neonatal period. All the survivors had severe developmental delay and multifocal neurological impairment. 92% developed refractory epilepsy, 58% were latter diagnosed with West syndrome. CONCLUSION: The non-reactive BS pattern may appear related to many neonatal neurological disorders and is associated with early and refractory neonatal seizures. It is clearly associated with elevated morbidity and mortality and to the development of post-neonatal epilepsy. PMID- 15830059 TI - Acoustic voice assessment in Parkinson's disease patients submitted to posteroventral pallidotomy. AB - Long-term complications in levodopa treated Parkinson's disease (PD) patients caused a resurgence of interest in pallidotomy as an option of treatment. However, postoperative complications such as speech disorders can occur. PURPOSE: The aim of this study is to evaluate the acoustic voice in PD patients, before and after posteroventral pallidotomy. METHOD: Twelve patients with PD were submitted to neurological and voice assessments during the off and on phases, in the pre-operative, 1st and 3rd post-operative months. The patients were evaluated with the UPDRS and the vocal acoustic parameters -- f0, NHR, jitter, PPQ, Shimmer, APQ (using the software MultiSpeech-Kay Elemetrics-3700). RESULTS: The off phase UPDRS scores revealed a tendency to improvement at the 1st month and the off phase worsened. The shimmer and APQ improved. CONCLUSION: This study shows that pallidotomy has little improvement on functional use of communication of PD patients. PMID- 15830060 TI - Status Dystonicus: study of five cases. AB - Status Dystonicus (SD) is characterized by generalized muscle contractions in dystonic patients. We report 5 cases of SD, two of which in patients with dystonic cerebral palsy, one in a patient with primary segmental dystonia, one in a patient with Hallervorden-Spatz syndrome and one in a patient with Wilson's disease (WD). Three patients were admitted to an intensive care unit and treated with propofol and midazolam, and two were submitted to neurosurgical procedures (bilateral pallidotomy and bilateral pallidal deep brain stimulation). Triggering factors were identified in three patients as follows: infection, stress-induced and zinc therapy for WD. On follow-up, two patients presented with significant improvement of dystonia, whereas the other three cases the clinical picture ultimately returned to baseline pre-SD condition. PMID- 15830061 TI - Botulinum toxin type A for the treatment of the upper limb spasticity after stroke: a meta-analysis. AB - Muscle over-activity is one of the cardinal features of spasticity and it is a common disability of stroke patients. In this group, spasticity is responsible for several limitations that interfere in their daily activities and quality of life. To treat spasticity, neurologists usually prescribe drugs as baclofen, tizanidine or benzodiazepines or even use definitive treatment as phenol or surgery. Authors suggest the use of botulinum toxin type A (BTX-A) for spasticity in the upper limbs after stroke, but there are few papers with adequate methodology supporting this idea. In this article we summarize the data of previous double-blind, randomised clinical trials to asses, with a meta-analysis, if BTX-A is an adequate treatment for spasticity due to stroke. The results show a statistical superiority of BTX-A ov%r placebo on reducing muscle tone by the Modified Ashworth Scale (WMD= 0.95 [0.74 to 1.17]) in patients with post-stroke upper limb spasticity. PMID- 15830062 TI - Does treatment with N-butyl cyanoacrylate embolization protect against hemorrhage in cerebral arteriovenous malformations? AB - OBJECTIVE: To assess the role of this procedure to prevent hemorrhage in cerebral arteriovenous malformations (cAVM). METHOD: Between 1992 and 2000, we studied 104 patients submitted to embolization as the main treatment. Patients were followed until hemorrhage or death. RESULTS: Follow-up ranged from 1.6 months to 8 years. The most frequent presentations were hemorrhage (50%) and seizures (38%). In addition, 40% were small (<30 mm); 56% were medium (30-60 mm). Obliteration was < or =1/3 in 11% of the cases; from 1/3 to < or = 2/3 in 49%; >2/3 in 36%; complete in 5%. The risk of death was 1%/year, and of bleeding, 5.4%/year. Presentation with hemorrhage and low obliteration rate were the main factors associated with hemorrhage. CONCLUSION: cAVM embolization provides limited protection against hemorrhage with obliteration rates below 2/3. Presentation with hemorrhage is the main factor for predicting hemorrhage. PMID- 15830063 TI - Intrinsic tectal low grade astrocytomas: is surgical removal an alternative treatment? Long-term outcome of eight cases. AB - Low-grade gliomas arising in dorsal midbrain in children and young patients usually present few neurological symptoms and findings, and patients management is controversial. Some authors propose only clinical observation until the patient present signs of increased intracranial pressure when a shunt with or without biopsy, is inserted; others recommend radiotherapy after stereotactic or open biopsy. Microsurgical total removal of tumor may be curative. We present a retrospective analysis of eight patients (mean age 16.6 +/- 11.5 years-old) with low-grade astrocytoma of the tectal region operated on using an infratentorial/supracerebellar approach between 1981 and 2002. All patients presented hydrocephalus and had a shunt insertion before surgical resection of the lesion. The tumour could be totally resected in seven patients. In one case radical removal was not possible due to infiltrative pattern of the lesion. Postoperative radiotherapy was performed in two cases, one patient at the beginning of this series and in the case with infiltrative tumor. This patient presented progressive tumor growth and died five years after surgery. No recurrence occurred after total removal. Post-surgical follow-up time ranged from 2 1/2 to 22 1/2 years (mean 9.9 +/- 5.9 years). Radical microsurgical removal of non invasive tumors is possible without mortality or significant morbidity. It may be curative and should remain as an alternative to be discussed with the patient. PMID- 15830064 TI - Para-muscular and trans-muscular approaches to the lumbar inter-vertebral foramen: an anatomical comparison. AB - Foraminal and extra-foraminal disc herniations comprise up to 11.7% of all lumbar disc herniations. Facetectomy, which had been the classic approach, is now recognized as cause of pain and instability after surgery. Otherwise, posterior lateral approaches through a trans-muscular or a para-muscular technique offer no significant damage to key structures for spinal stability. The surgical anatomy of these approaches has already been described, but they were not compared. In order to quantify the angle of vision towards the intervertebral foramen offered by each technique, 12 fresh cadavers were dissected and studied regarding these approaches. The angle presented by trans-muscular approach was wider in all studied lumbar levels. Surgery through the trans-muscular approach is performed with a better working angle, requiring a smaller resection of surrounding tissues. Therefore, minor surgical trauma can be expected. Our measurements support previously published data that point the trans-muscular approach as the best surgical option. PMID- 15830065 TI - Combined supra/infratentorial approach to tentorial meningiomas. AB - In spite of significant advancements in imaging technology, monitoring, and microsurgical techniques, complete and safe removal of tentorial meningiomas remains a challenge for most neurosurgeons. Classifications of tentorial meningiomas are revised. The combined supra/infratentorial approach to resect tentorial meningioma is discussed. This approach provides a wider exposure of the supra/infratentorial region with less brain retraction. With this approach the occipital lobe and the cerebellum are exposed along the tentorium. Two illustrative cases are presented. The patients were studied with computerized tomography, magnetic resonance and angiography. The anatomy of the transverse sinus and the confluence of the sinus could be appreciated with these studies. The operative technique is described stepwise. Emphasis is placed on pre operative evaluation and surgical technique, leading to a total surgical removal of the lesion with margins of safety. The goal of surgical treatment of tentorial meningiomas is their complete and safe removal. With this unique approach we sought to confirm that it offers a safe means of resection not only the neoplasm but also the infiltrated dura. PMID- 15830066 TI - [Intraoperative mapping of motor areas during brain tumor surgery: electrical stimulation patterns]. AB - Brain mapping with direct electrical stimulation is usefull when the tumor is located near or has infiltrated the central lobe. OBJECTIVE: To analyze the surgical findings with direct electrical stimulation of the cortex and white matter under general anesthesia during surgery for brain tumors related to the central lobe. METHOD: We studied 42 patients operated on from June 2000 to June 2003. We analyzed surgical findings and details of brain mapping. RESULTS: The mean value of the intensity of the stimulus was greater among those who presented motor deficit prior to surgery (p = 0.0425) and edema on MRI (p = 0.0468) or during anesthesia with continuous propofol (p = 0.001). CONCLUSION: The functional mapping of the central lobe may be influenced by severe motor deficit, edema on MRI and propofol's anesthesia. PMID- 15830067 TI - [Idiopathic childhood occipital epilepsies: clinical and electroencephalographic features in 63 children]. AB - We studied clinical-EEG features of 63 children, age range of 2-14 years, with occipital epileptiform activity in the EEG, with epileptic seizures and no evidences of brain damage. Age at onset was between 1-12 years. In 15 cases the seizures last for more than 30 min up to 6 h. Seizures occurred during sleep in 28 children. Autonomic symptoms were apparent in 25, head deviation in 24 and hemiconvulsion or generalization in 17 cases. Ten children reported visual symptoms. In 14 children the seizures were exclusively generalised convulsions. Spikes were observed in 27 cases and spike and slow-wave complex in 36. Discharges blocking by eyes opening were confirmed in 15 cases. Somatosensory evoked spikes by foot stimulation were observed in 9.5% cases. The cases were classified in the following idiopathic epileptic syndromes: early-onset occipital -- Panayiotopoulos-type (EOO), 32 cases; late-onset occipital -- Gastaut type (LOO), 8; rolandic epilepsy in 7. CONCLUSION: EOO occurred more frequently than LOO. Idiopathic occipital syndromes diagnosis was possible in 63.4% of the cases. PMID- 15830068 TI - [Contribution of psychosocial risk factors for attention deficit/hyperactivity disorder]. AB - OBJECTIVE: To analyze the contribution of psychosocial risk factors for the diagnosis of attention deficit/hyperactivity disorder (ADHD). METHOD: This is a retrospective, case-control study that analyzed 403 pupils from a public elementary school. DSM-IV symptom questionnaire and a specific psychosocial questionnaire were filled out, and a history and complete physical and neurological examinations were performed. RESULTS: The most statistically significant psychosocial risk factor was "marital discord in the past" (odds ratio = 11.66, p = 0.044). A total of 29 murders of subjects' close relatives were observed, and this variable was statistically relevant. CONCLUSION: Children that were exposed to marital discord in the past between parents underwent a 11.66-fold increase in the risk of ADHD diagnosis. Authors suggest that exposure to community violence be included among Rutters's indicators of adversity, which predispose infants and children to the future occurrence of ADHD and other neuropsychiatric disorders. PMID- 15830069 TI - [Neurodevelopment of full-term small-for-gestational age infants in the second month of life]. AB - The objective of the present study was to assess and to compare the neurodevelopment of full-term adequate (AGA) or small-for-gestational age (SGA) infants in the second month of life. Sixty-seven infants were evaluated: 43 AGA and 24 SGA, making use of the Bayley Scales of Infant Development. The SGA group Index Score (IS) was significantly lower in Mental and Motor Scales. Considering the body proportionality (Asymmetric, Symmetric-SGA and Control group) there was difference in Motor Scale (p=0.003) with lower scores in the Symmetric-SGA group. Comparing to the Control group IS percentiles, in Mental Scale there was difference between Asymmetric X Symmetric-SGA; in Motor Scale, there was difference between the Asymmetric X Control (p=0.039) and Symmetric-SGA X Control (p=0.007) groups; there was no difference between Asymmetric and Symmetric-SGA although both exhibited lower scores than the Control group. PMID- 15830070 TI - [EEG alpha band coherence analysis in healthy adults: preliminary results]. AB - We studied the occipital inter-hemispheric coherence of Electroencephalogram (electrodes O1-O2) for alpha band (alpha1--8.0 to 10.0 Hz and alpha2--10.1 to 12.5 Hz) in two groups of healthy individuals (young adults and subjects older than 50 years-old), to assess if there is significant difference between this two age groups. No significant difference in alpha band coherences was found between these two age groups. PMID- 15830071 TI - [Cefepime-induced encephalopathy: clinical and electroencephalographic features in seven patients]. AB - Cefepime, a fourth-generation cephalosporin, with large antibacterial spectrum, is a commonly used antibiotic for the treatment of serious hospital infections. Its security report is considered favourable. Recently, many cases of a severe and reversible cefepime-induced encephalopathy were described. In this paper, we report seven patients with reversible cefepime-induced encephalopathy, with a peculiar EEG pattern, characterized by semiperiodic diffuse triphasic waves. We discuss the EEG abnormalities found and their association with nonconvulsive status epilepticus. PMID- 15830072 TI - [Pitfalls in the clinical and electroencephalographic diagnosis of ceroid lipofuscinosis]. AB - Neuronal ceroid lipofuscinosis (NCL) were traditionally classified according to age of onset and clinical features in four main groups. Recently, a combination of clinical, ultra structural and genetics data led to the recognition of eight forms of NCL, providing a more precise framework to classify atypical cases. By the other hand, it was shown that mutations in the same gene could be responsible for a large variety of clinical phenotypes. The objective of this study is to describe two brothers with clinical and electroencephalographic abnormalities characteristic of juvenile NCL, but with ultra structural abnormalities suggestive of late infantile NCL. Electroencephalogram is useful for clinical diagnosis of NCL but it is not helpful in its classification. PMID- 15830073 TI - [Association between plasmatic levels of homocysteine and stroke: a transversal analytic study]. AB - PURPOSE: To determine the association of homocysteine with ischemic stroke, considering age, gender, cigarette smoking, hypertension, diabetes and etiology of cerebrovascular disease. METHOD: Transversal analytic observational study of 104 patients with the diagnosis of ischemic stroke and 98 healthy controls had blood homocysteine analyzed by high performance liquid chromatography. Statistics was performed with Student's t and Kruskal-Wallis' tests, analysis of variance, linear regression analysis, logistic regression and Pearson's linear correlation coefficient. RESULTS: Patients had higher values for homocysteine (15.4 +/- 11.7 micromol L(-1)) than controls (10.5 +/- 4.2 micromol L(-1)). Dividing both groups into four groups according to age, significant differences in homocysteine values were found between patients and controls with age 40-49 and 50-59 years. Homocysteine values were significantly higher in patients with hypertension and in those with diagnosis of atherosclerotic cerebrovascular disease. Logistic regression studies showed relationship between atherosclerosis and homocysteine. CONCLUSIONS: Homocysteine is associated with ischemic stroke. PMID- 15830074 TI - [Electrophysiological study of the central and peripheral hearing system of aphasic individuals]. AB - SUBJECT: Electrophysiology of the auditory system. OBJECTIVE: Electrophysiological evaluation of the peripheral and central auditory system of brain injured patients. METHOD: Experimental group: eleven brain injured and aphasic subjects, both genders and with ages ranging from 43 to 75; control group: eleven individuals without hearing complaints, equalized as to gender and age. The subjects were evaluated through auditory brainstem response (ABR); auditory middle latency response (AMLR) and auditory P300 response. RESULTS: An increase in the V wave latency and I-V interpeak in both groups, due to the age factor. The presence of statistically significant hemispheric differences when compared to the Pa component in MLAEP research, registered in the C3 (left hemisphere) and the C4 (right hemisphere). In researching the P300 Cognitive Potential, there was an absence or increase in P300 latency and a decrease in P300 amplitude in the presence of the N2 component. CONCLUSION: The AMLR and auditory P300 response have proven to be effective instruments for evaluating aphasic individuals. PMID- 15830075 TI - [Influence of the respiratory physiotherapy on intracranial pressure in severe head trauma patients]. AB - OBJECTIVE: To evaluate influence of the respiratory physiotherapy on intracranial pressure (ICP) in patients with severe head trauma. METHOD: Thirty five patients with severe head trauma were included in the study. The patients were divided into three groups: ICP 0-10, 11-20 and 21-30 mmHg. The following variables were measured: ICP and mean arterial pressure. Cerebral perfusion pressure was calculated as the difference between mean arterial and intracranial pressure. RESULTS: Endotracheal aspiration increased ICP in all patients. The mean arterial pressure didn't change and cerebral perfusion pressure decreased, but remaining normal value. CONCLUSION: Respiratory physiotherapy maneuvers can be safely applied in patients with severe head trauma with ICP below 30 mmHg. More attention should be taken during endotracheal aspiration. PMID- 15830076 TI - [Awareness of disease in dementia: preliminary results in patients with mild and moderate Alzheimer's disease]. AB - OBJECTIVE: To evaluate the presence and the level of awareness of disease in mild and moderate Alzheimer's disease (AD). METHOD: Cross-sectional evaluation of patients with mild/moderate AD (n=42) assessed by Assessment of Psychosocial Impact of the Dementia Diagnosis (APSID), Mini-mental State Examination (MMSE) and Clinical Dementia Rating Scale (CDR). RESULTS: Awareness of disease and its consequences were present in 66.7% patients with mild AD (n=18). In moderate AD (n=24), 20.8% presented total awareness, 45.8% presented only awareness of cognitive symptoms. Unawareness of disease was observed in 33.3%. CONCLUSIONS: The present data show association between awareness and level of severity of disease. CDR 1 patients show a better recognition of cognitive and daily life activity symptoms, whereas CDR 2 patients recognized their cognitive symptoms but failed to appraise their severity and consequences in daily life activities. PMID- 15830077 TI - [Influence of education on the neuropsychological assessment of the elderly: application and analysis of the results from the Mattis Dementia Rating Scale (MDRS)]. AB - OBJECTIVE: To assess the influence of low education and illiteracy on the evaluation of dementia by the Mattis Dementia Rating Scale (MDRS). METHOD: We applied the MDRS to 62 normal elderly subjects (64-77 years), divided into five groups according to schooling, i.e., 15-16 years, 11-12 years, 8-9 years, 4 years, and illiterate. The MDRS covers the study of five subscales and the sum of their scores may represent the degree of cognitive impairment. RESULTS: A significant difference (p < 0.05) in performance on this scale was observed among the subgroups in relation to education on 12 items, 5 subscales (illiterates < all other groups and 15-16 years > 4 and 8 years of schooling, p < or = 0.001) and the total MDRS score (illiterates < all others and 4 and 8 years < 15-16 years, p < 0.001). No significant correlations where found for age and, regarding gender, the differences were significant in only one item. CONCLUSION: Education interfered with individual performance on the MDRS. Illiteracy is a determinant factor of lower MDRS scores that could generate diagnostic errors. PMID- 15830078 TI - [Multiple Sclerosis Functional Composite Measure (MSFC) standardized in the Brazilian population]. AB - The Multiple Sclerosis Functional Composite Measure (MSFC) is an outcome measure in multiple sclerosis developed by USA National Multiple Sclerosis Society (1994), a three-part composite clinical measure--9-Hole Peg Test, Timed 25-Foot Walk and PASAT. It should be multidimensional in order to reflect the principal ways MS affects an individual. The MSFC was applied in 91 Brazilian subjects and standardized to be use in MS centers. PMID- 15830079 TI - [Ethical issues in multiple sclerosis under physicians and patients point of view]. AB - Multiple sclerosis (MS) is a neurologic disorder that mostly affects young adults and can usually evolute to physical disability. Thus, caring patients with MS brings many ethic questions for the physician. OBJECTIVE: To identify physicians and patients' perceptions about the illness and so improve doctor-patient relationship. METHOD: It was made two different questionnaires, one for patients and another for physicians, 103 patients and 44 physicians answered them. RESULTS: 96.1% of patients knew their diagnosis, all others would like to know it. From those, 74.7% thought that that way it was disclosured was correct and 90.9% said that the doctor should tell us it. The worst symptoms described were fatigue (29.1%) and motor deficits (28.1%). By other side, 68% of patients told they suffered because of the illness. The most important reason for doctors to tell the diagnosis to the patients was to improve adherence to treatment (56.8%). A familiar present at this moment was demanded for 54.6% of doctors. When asked about orientations in a pregnancy, 50% of physicians did not answer correctly. Finally, 50% of physicians were against complementary and alternative therapies. CONCLUSION: Patients want to know their diagnosis and doctors should tell them in the most adequate moment and give more information. A debate about palliative care is also necessary. PMID- 15830080 TI - [Diagnosis and treatment of the most frequent vestibular syndromes]. AB - The aims of this study were to identify the most common vestibular syndromes in a dizziness unit, and to observe their clinical aspects and response to treatment. Five hundred and fifteen patients were studied retrospectively in two institutions. Aspects of anamnesis, physical examination and the response to treatment were evaluated. The most frequent syndromes were: benign paroxysmal positioning vertigo (VPPB) (28.5%), phobic postural vertigo (11.5%), central vertigo (10.1%), vestibular neuritis (9.7%), Meniere disease (8.5%), and migraine (6.4%). A good response to treatment was observed in most patients with migraine (78.8%), VPPB (64%), vestibular neuritis (62%), Meniere disease (54.5%) and vestibular paroxismia (54.5%). On the other hand, patients with downbeat nystagmus and bilateral vestibulopathy had poor response (52.6% and 42.8%, respectively). The diagnosis of these most frequent vestibular syndromes were established through anamnesis and physical examination (with specific clinical tests for evaluation of the vestibular function). The correct diagnosis and adequate treatment are important since these syndromes may have a good prognosis. PMID- 15830081 TI - [Diagnostic difficulties in spinal muscular atrophy]. AB - OBJECTIVE: To describe the clinical findings of patients with spinal muscular atrophy (SMA) with survival motor neuron (SMN) gene deletion. METHOD: Descriptive study of SMA cases confirmed with the deletion of the SMN gene. Frequency determination of positive clinical and laboratory revised diagnostic criteria. RESULTS: All of the 22 included patients had symmetrical muscle weakness, which was diffuse in those with onset of symptoms up to 6 months of age (75 %), and either proximal or predominant in lower limbs in the remaining group (67 %). Fasciculations and atrophy were both frequent findings (82 %). Laboratory tests findings were variable, with a positivity of 57 % for electrophysiology and of 58 % for muscle biopsy. CONCLUSION: The presence of a deletion in the SMN gene can help to confirm this diagnosis in unclear presentations. PMID- 15830082 TI - Head tremor and progressive multifocal leukoencephalopathy in AIDS patients: report of two cases. AB - Progressive multifocal leukoencephalopathy (PML) is caused by replication of JC virus in oligodendrocytes of immunocompromised patients. Common manifestations are focal motor and sensory deficits, gait abnormalities, speech and language disturbances, cognitive disorders, headache, and visual impairment. Although the occurrence of movement disorders is rare in PML, bradykinesia, rigidity, dystonia, myoclonic jerks and myoclonic ataxia have been described. Head tremor associated with PML has not been previously reported. We report two cases of PML in whom head tremor was present. PMID- 15830083 TI - X-linked spinal and bulbar muscular atrophy (Kennedy's disease) with long-term electrophysiological evaluation: case report. AB - X-linked spinal and bulbar muscular atrophy or Kennedy's disease is an adult onset motor neuronopathy caused by a CAG repeat expansion within the first exon of an androgen receptor gene. We report the case of a 66-year-old man, previously diagnosed with motor neuron disease (MND), who presented acute and reversible left vocal fold (dysphonia) and pharyngeal paresis, followed by a slowly progressive weakness and also bouts of weakness, wasting and fasciculation on tongue, masseter, face, pharyngeal, and some proximal more than distal upper limb muscles, associated to bilateral hand tremor and mild gynecomastia. There were 5 electroneuromyography exams between 1989 and 2003 that revealed chronic reinnervation, some fasciculations (less than clinically observed) and rare fibrillation potentials, and slowly progressive sensory nerve action potentials (SNAP) abnormality, leading to absent/low amplitude potentials. PCR techniques of DNA analysis showed an abnormal number of CAG repeats, found to be 44 (normal 11 34). Our case revealed an acute and asymmetric clinical presentation related to bulbar motoneurons; low amplitude/absent SNAP with mild asymmetry; a sub-clinical or subtle involvement of proximal/distal muscles of both upper and lower limbs; and a probable evolution with bouts of acute dennervation, followed by an efficient reinnervation. PMID- 15830084 TI - Primary hyperparathyroidism simulating motor neuron disease: case report. AB - We report a case of a 26-year-old man who presented a lower motor neuron syndrome due to hyperparathyroidism. Electromyography showed neurogenic features with normal nerve conduction studies. Hypercalcemia led to the discovery of a primary hyperparathyroidism with gland hyperplasia. Following parathyroid surgery there was recovery of the neurological symptoms. PMID- 15830085 TI - [Cavernous hemangioma of the internal auditory canal encasing the VII and VIII cranial nerve complex: case report]. AB - We report the surgically treated case of a 21-year-old caucasian male harboring a cavernous hemangioma of the right internal auditory canal encasing the seventh and eighth cranial nerves complex. Only 18 cases of cavernous hemangiomas of this location have been previously reported. The clinical features, the differential diagnosis and the treatment are discussed. PMID- 15830086 TI - Multiple meningiomas within the spinal canal: case report with 23 tumors. AB - In most series of neoplasms of the spinal canal, spinal cord and its leptomeninges, the incidence of meningioma group comprises approximately 25%. The incidence of multiple meningiomas is small when compared with the frequency of its single occurrence. In the majority of cases, their multiplicity is intracranial and spinal. Multiple meningiomas confined exclusively to the spinal canal are extremely rare. We report on a 33 years-old man, with 23 tumors located in the spinal thoracic region. PMID- 15830087 TI - [Transient lesion in the splenium of the corpus callosum in epileptic child with cerebral low grade glioma]. AB - We report on a seven years-old boy with complex partial seizures and the presence of low grade glioma in left fronto-parietal region. The magnetic resonance imaging showed focal non-hemorrhagic lesion in the splenium of the corpus callosum. The description of the transient lesion in the splenium of the corpus callosum was related in three previous studies, in patients with epilepsy. Thus, the observed transient focal lesion in the splenium of the corpus callosum of this child, probably, has correlation with to prolonged focal partial seizures and not to the presence of glioma low grade. PMID- 15830088 TI - [Ophthalmoplegic migraine: MRI findings. Case report]. AB - Ophthalmoplegic migraine is a rare syndrome in which headache is associated with ophthalmoplegia and third, fourth or sixth cranial nerves palsy. It occurs most frequently in childhood and teenagers. At magnetic resonance imaging (MRI) with gadolinium (GD-DTPA) it may be observed a transitory enhancement of the affected nerve. We present the case of a male teenager, 16 years old, with typical medical history and enhanced signal at left oculomotor nerve in cisternal portion at MRI weighted in T1 with GD-DTPA. On the control exam, eighteen months later, there was no remarkable lesion. The enhancement of oculomotor nerve at MRI is always pathological and among the differential diagnosis we must include: neoplasia (lymphoma and leukemia), infections (AIDS, syphilis), inflammatory process (sarcoidose and Tolosa-Hunt syndrome) and vascular (posterior communicating artery aneurysm). PMID- 15830089 TI - [Wilson's disease: clinical diagnosis and "faces of panda" signs in magnetic resonance imaging. Case report]. AB - A 25 year-old man was admitted with polymorph symptomatology resembling basal ganglia disease associated with psychiatric manifestations. The patient had been treated with pericyazine. The drug was stopped but the symptomatology did not improve. The diagnosis of Wilson's disease was established through ophthalmologic examination with slit-lamp that revealed the Kayser-Fleischer ring and laboratory abnormalities showing a low serum ceruloplasmin level and increased urinary copper excretion. T2-weighted axial magnetic resonance imaging demonstrated the " face of panda signs" in the midbrain and pons. PMID- 15830090 TI - Botulinum toxin: mechanisms of action. AB - This review describes therapeutically relevant mechanisms of action of botulinum toxin (BT). BT's molecular mode of action includes extracellular binding to glycoproteine structures on cholinergic nerve terminals and intracellular blockade of the acetylcholine secretion. BT affects the spinal stretch reflex by blockade of intrafusal muscle fibres with consecutive reduction of Ia/II afferent signals and muscle tone without affecting muscle strength (reflex inhibition). This mechanism allows for antidystonic effects not only caused by target muscle paresis. BT also blocks efferent autonomic fibres to smooth muscles and to exocrine glands. Direct central nervous system effects are not observed, since BT does not cross the blood-brain-barrier and since it is inactivated during its retrograde axonal transport. Indirect central nervous system effects include reflex inhibition, normalisation of reciprocal inhibition, intracortical inhibition and somatosensory evoked potentials. Reduction of formalin-induced pain suggests direct analgesic BT effects possibly mediated through blockade of substance P, glutamate and calcitonin gene related peptide. PMID- 15830091 TI - [Temporal dementia]. PMID- 15830093 TI - Hyperhomocysteinemia and atherosclerosis. AB - Arteriosclerosis and its complications, such as heart attack and stroke, are the major causes of death in developed countries. It was believed that age, hyperlipidemia, hypertension, diabetes and smoking are common risk factors for cardiovascular disease. In addition, overwhelming clinical and epidemiological studies have identified homocysteine (Hcy) as a significant and independent risk factor for cardiovascular disease. In healthy individuals, plasma Hcy is between 5 and 10 micromol/L. One cause of severe hypehomocys- teinemia (HHcy) is the deficiency of cystathionine beta-synthase (CBS), which converts Hcy to cystathionine. CBS homozygous deficiency results in severe HHcy with Hcy levels up to 100 to 500 micromol/L. Patients with severe HHcy usually present with neurological abnormalities, premature arteriosclerosis. It has been reported that lowering plasma Hcy improved endothelial dysfunction and reduced incidence of major adverse events after percutaneous coronary intervention. The mechanisms by which Hcy induces atherosclerosis are largely unknown. Several biological mechanisms have been proposed to explain cardiovascular pathological changes associated with HHcy. These include: (1) endothelial cell damage and impaired endothelial function; (2) dysregulation of cholesterol and triglyceride biosynthesis; (3) stimulation of vascular smooth muscle cell proliferation; (4) thrombosis activation and (5) activation of monocytes. Four major biochemical mechanisms have been proposed to explain the vascular pathology of Hcy. These include: (1) autooxidation through the production of reactive oxygen species; (2) hypomethylation by forming SAH, a potent inhibitor of biological transmethylations; (3) nitrosylation by binding to nitric oxide or (4) protein homocysteinylation by incorporating into protein. In summary, our studies, as well as data from other laboratories support the concept that Hcy is causally linked to atherosclerosis, and is not merely associated with the disease. Although folic acid, vitamin B12 and B6 can lower plasma Hcy levels, the long term effects on cardiovascular disease risk are still unknown and judgments about therapeutic benefits await the findings of ongoing clinical trials. PMID- 15830094 TI - Bicarbonate secretion by the female reproductive tract and its impact on sperm fertilizing capacity. AB - The luminal fluid environment of the female reproductive tract is considered critical for the sperm to undergo a series of molecular events leading to the final acquisition of their fertilizing capacity. It has been shown that the fluid in the female reproductive tract contains high content of HCO3- and it plays an important role in sperm functions including sperm motility, capacitation, hyperactivation and acrosome reaction. This review summarizes the effects of HCO3 on sperm functions occurring in the female reproductive tract and discusses the transport mechanisms involved in mediating uterine HCO3- secretion. New evidence is also presented to show possible cause of female infertility due to defective HCO3- transporting mechanism. PMID- 15830095 TI - [Effects of rosiglitazone on aortic function in rats with insulin resistant hypertension]. AB - Rosiglitazone (ROSI), thiazolidione peroxisome proliferator-activated receptor gamma (PPAR-gamma) activator, reduces insulin resistance in patients with type 2 diabetes (T2DM). It also improves vascular reactivity in T2DM patients and some animal models by unclear mechanisms. In order to investigate the effect of ROSI on aortic systolic and diastolic function of insulin resistant-hypertensive rats (IRHR) and the underlying mechanism, male Sprague-Dawley (SD) rats were fed with high fructose (HF) for 8 weeks to induce IRHR model. To verify IRHR model, systolic blood pressure (SBP), fasting blood sugar (FBS), fasting serum insulin (FSI) were measured respectively in each group, and insulin sensitive index (ISI) was also calculated. Subsequently, the vascular function test was performed. The thoracic aortic ring of SD rats was mounted on a bath system. The effect of rosiglitazone on the contraction elicited by L-phenylephrine (PE) and potassium chloride (KCl) and the relaxation induced by acetylcholine (ACh) and sodium nitroprusside (SNP) were measured. To explore the mechanism, nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) was used and serum nitric oxide (NO) was measured. The results obtained were as follows: (1) Rosiglitazone reduced the level of SBP, serum insulin and improved insulin resistance in IRHRs. (2) The contractive responses of thoracic aortic rings to PE and KCl were enhanced and the relaxation response to ACh was depressed significantly in the HF group, and the effect was reversed by ROSI. (3) After pretreatment with L-NAME, the relaxation response to ACh was further impaired in the HF group, this effect was partly reversed by ROSI. (4) Sodium nitroprusside (SNP)-induced vasodilator responses did not differ significantly among the groups. (5) Aortic systolic and diastolic function of the control group was not affected markedly by ROSI. (6) Compared with the control group, serum nitric oxide was significantly reduced in the HF group, but after rosiglitazone treatment it was remarkably increased. These findings suggest that ROSI can improve aortic diastolic function of insulin resistant-hypertensive rats, the mechanism of this effect might be associated with an increase in nitric oxide mediated partly by NOS pathway, a decrease in the level of blood pressure, serum insulin and the improvement of insulin resistance. PMID- 15830096 TI - Induction of rat neural stem cells into oligodendrocyte precursor cells. AB - We have previously established a culture method to isolate and cultivate neural stem cells (NSCs) derived from the rat embryonic brain and spinal cord. In the present study, we demonstrate that the spinal cord-derived NSCs can be induced to differentiate into oligodendrocyte precursor cells (OPCs) with a combined treatment composed of (1) conditioned medium collected from B104 neuroblastoma cells (B104CM) and (2) basic fibroblast growth factor (bFGF, 10 ng/ml). After induction, over 95% of the cells displayed bipolar or tri-polar morphology and expressed A2B5 and platelet derived growth factor receptor-alpha (PDGFR-alpha), markers that are specific for OPCs. Among PDGFR-alpha positive OPCs, only a few cells expressed glia fibrillary acidic protein (GFAP) and none expressed beta tubulin III. In the presence of B104CM and bFGF, OPCs proliferated rapidly, formed spheres, expanded for multiple passages, and maintained their phenotypic properties. Upon withdrawal of B104CM and bFGF, these cells differentiated into either O4/GlaC-positive oligodendrocytes (OLs) or GFAP- and A2B5-positive type-2 astrocytes. Our results indicate that NSCs can be induced to differentiate into OPCs that possess properties of self-renewal and differentiation into oligodendrocytes and type-2 astrocytes, a property similar to that of O-2A progenitor cells. The OPCs can be maintained in an undifferentiated state over multiple divisions as long as both B104CM and bFGF are present in the medium. Thus, large quantity of OPCs can be obtained through this method for potential therapeutical interventions for various neurological degenerative diseases. PMID- 15830097 TI - [cAMP response-element binding protein participates in the phosphorylated extracellular signal-regulate kinase mediated neuropathic pain]. AB - It has been reported that extracellular signal-regulate kinase (ERK) is involved in the modulation of nociceptive information and central sensitization produced by intense noxious stimuli and/or peripheral tissue inflammation. Few studies have explored the relationship between ERK and cAMP response-element binding protein (CREB) in neuropathic pain after nerve injury, such as chronic constriction injury (CCI) of the sciatic nerve. In the present study, CCI model was employed to investigate the activation of ERK on the expression of phosphorylated CREB (pCREB) in chronic neuropathic pain. Lumbar intrathecal catheters were chronically implanted in male Sprague-Dawley rats. The left sciatic nerve was loosely ligated proximal to the sciatica's trifurcation at around 1.0- mm intervals with 4-0 silk suture. Mitogen-activated protein kinase kinase (MEK) inhibitor U0126 and phosphorothioate-modified antisense oligonucleotides (ODN) were intrathecally administered one day before and three consecutive days after CCI. Thermal and mechanical nociceptive thresholds were assessed with the paw withdrawal lantency (PWL) to radiant heat and von Frey filaments respectively. The expression of pCREB and Fos were assessed by both Western blot and immunohistochemical analysis. The results showed that intrathecal injection of U0126 or ERK antisense ODN attenuated significantly CCI induced mechanical and thermal hyperalgesia. Correlating with behavior results, the injection also markedly suppressed the increase of CCI-induced pCREB and c Fos expression. The results obtained suggest that CREB participates in the pERK mediated neuropathic pain. PMID- 15830098 TI - [Effect of tetramethylpyrazine on the responses of respiration and expression of nNOS in brainstem to hypoxia in rats]. AB - The aim of the present study was to investigate the effect of tetramethylpyrazine (TMP) on the changes of respiration and expression of neuronal nitric oxide synthase (nNOS) in brainstem induced by hypoxia in the rats. Hypoxia was induced by inhalation of 8% O2-balanced N2.The electromyogram (EMG) of diaphragm was monitored to evaluate the respiratory response of the rats to hypoxia. The immunohistochemical staining technique was used to study the change of the expression of nNOS in the brainstem during hypoxia. In the rats of hypoxia group, a successive process of response, excitatory followed by inhibitory, was produced. Twenty min after hypoxia, a significant inhibition of respiration occurred, which was characterized with a marked decrease in the inspiratory duration, the respiratory frequency, and the amplitude of inspiration and a prolongation of expiratory duration (P<0.05). In the rats of pretreated with TMP, the respiratory activity was not obviously depressed (P>0.05). In the rats of hypoxia group, the level of nNOS immunoreactivity was enhanced remarkably in the lateral reticular nucleus, nucleus of trapezoid, hypoglossal nucleus and the facial nucleus compared with the control group (P<0.05). In the rats of pretreated with TMP, the nNOS level increased further in the nuclei mentioned above (P<0.05). The results obtained indicate that TMP can reverse the inhibitory effect of hypoxia on respiration in the rats and that nNOS may be involved in the respiratory protective action of TMP. PMID- 15830099 TI - [Ginsenoside Rb1 attenuates okadaic acid-induced Tau protein hyperphosphorylation in rat hippocampal neurons]. AB - The present study was aimed to investigate the effects of ginsenoside Rb1 on okadaic acid (OA)-induced Tau hyperphosphorylation in hippocampal neurons of Sparague-Dawley rat and to explore its possible mechanism. Animals were randomly divided into four groups. Group 1 received dimethysulphoxide (DMSO) injection (vehicle group), group 2 only received OA injection (OA group), group 3 was pretreated with Rb1 and then received OA injection (Rb1 pretreatment group), and the group 4 was an intact control group. The animals in group 3 were injected intraperitoneally with various doses of Rb1 at 5, 10, and 20 mg/kg (once a day for 14 d). On the thirteen day of pretreatment, animals in Rb1 pretreatment group as well as animals in OA group received a bolus injection of 0.483 microg of OA (1.5 microl of solution in DMSO) at right dorsal aspect of hippocampus to induce Tau hyperphosphrylation. The brains were harvested one day after the last treatment. In all groups, the morphology of neurofibrils, phosphorylation of Tau protein, and the activity of phosphatase 2A (PP2A) were investigated. In OA group, the Bielschowski's assay revealed darkened and uneven neurofibrils staining in the hippocampus. The immunohistochemistry results showed a significant increase in Thr(231) phosphorylation of Tau protein in OA group relative to the control group (P<0.01). OA injection also markedly decreased PP2A activity (P<0.01). Western blot confirmed Thr(231) phosphorylation of Tau protein and it also detected phosphorylation of Ser(396) of Tau protein. The animals with Rb1 pretreatment displayed even staining of neurofibrils and normal pattern of fiber organization. Rb1 pretreatment also attenuated Thr(231) and Ser(396) hyperphosphorylations of Tau protein, and restored PP2A activity compared to the OA group (P<0.01). These results indicate that OA-induced hyperphosphorylation of Tau protein in rat hippocampal neurons can be attenuated by the pretreatment of ginsenoside Rb1. These data also implicate that Rb1 has potential neuroprotective effects on Tau-related neuropathology. PMID- 15830100 TI - [Different roles of the spinal protein kinase C alpha and gamma in morphine dependence and naloxone-precipitated withdrawal]. AB - Our previous studies showed that spinal neurons sensitization was involved in morphine withdrawal response. This study was to investigate the roles of spinal protein kinase C (PKC) alpha, gamma in morphine dependence and naloxone precipitated withdrawal response. To set up morphine dependence model, rats were subcutaneously injected with morphine (twice a day, for 5 d). The dose of morphine was 10 mg/kg in the first day and was increased by 10 mg/kg each day. On day 6, 4 h after the injection of morphine (50 mg/kg), morphine withdrawal syndrome was precipitated by an injection of naloxone (4 mg/kg, i.p.). Chelerythrine chloride (CHE), a PKC inhibitor, was intrathecally injected 30 min before the administration of naloxone. The scores of morphine withdrawal symptom and morphine withdrawal-induced allodynia were observed. One hour after naloxone precipitated withdrawal, Fos protein expression was assessed by immunohistochemical analysis and Western blot was used to detect the expression of cytosol and membrane fraction of PKC alpha and gamma in the rat spinal cord. The results showed that intrathecal administration of CHE decreased the scores of morphine withdrawal, attenuated morphine withdrawal-induced allodynia and also inhibited the increase of Fos protein expression in the spinal cord of morphine withdrawal rats. The expression of cytosol and membrane fraction of PKC alpha was significantly increased in the spinal cord of rats with morphine dependence. Naloxone-precipitated withdrawal induced PKC alpha translocation from cytosol to membrane fraction, which was prevented by intrathecal administration of CHE. During morphine dependence, but not naloxone-precipitated withdrawal, PKC gamma in the spinal cord translocated from cytosol to membrane fraction, and intrathecal administration of CHE did not change the expression of PKC gamma in the spinal cord of naloxone-precipitated withdrawal rats. It is suggested that up regulation and translocation of PKC in the spinal cord contribute to morphine dependence and naloxone-precipitated withdrawal in rats and that PKC alpha and gamma play different roles in the above-mentioned effect. PMID- 15830101 TI - [Effect of microinjection of atrial natriuretic peptide into the paraventricular nucleus on baroreflex sensitivity in conscious rats]. AB - The role of atrial natriuretic peptide (ANP) in the central regulation of the circulation is known to be a neurotransmitter or a neuromodulator, but its actions on baroreceptor reflex function are not fully resolved. The present study examined the role of ANP (6, 60 ng/0.2 microl) by direct microinjection into the hypothalamic paraventricular nucleus (PVN) in conscious rats. OPC-21268 (0.45 microg/3 microl), an antagonist of the V(1) receptor, was microinjected into the lateral ventricle to examine whether the effect of ANP on baroreflex sensitivity is mediated by vasopressin (VP). ANP significantly increased the baroreflex sensitivity, and OPC-21268 attenuated the increase of baroreflex sensitivity induced by ANP. Intravenous injections of ANP (60 ng/0.04 ml) did not affect baroreflex sensitivity. These results suggest that ANP in the PVN may produce a facilitative effect on baroreflex, and the effect may be via, at least in part, the central vasopressin. PMID- 15830102 TI - Spontaneous discharges of pyramidal cells in the dorsal hippocampus of guinea pig. AB - To determine the characteristics of spontaneous discharges of hippocampal pyramidal cells (PCs), extracellular neuronal firing in CA1 and CA3 regions of dorsal hippocampus was recorded, the firing modes and interspike interval (ISI) were analyzed with the conventional and nonlinear methods. PCs were discriminated from interneurons using the measurement of action-potential duration and firing rate in this study. There was no significant difference in duration, mean firing frequency, complexity and firing mode between the neurons in CA1 and CA3 regions both in anesthetized and awake animals. The complexity of neurons was higher in awake group than that in anesthetized group, though no difference was found in firing rate. There were differences in the type of pyramidal cells and the coefficient of variance of ISI of neurons. The results obtained from the spontaneous discharges of dorsal hippocampal pyramidal cells reveal some nonlinear and linear aspects in anesthetized and awake states. It seems likely that the combination of conventional and non-linear measurements of the hippocampal pyramidal cells encoding may reflect genuine characteristics of the hippocampal pyramidal cells. PMID- 15830103 TI - [Regulatory role of HTm4 gene in hematopoietic cell cycle]. AB - Cell cycle progression is tightly regulated in hematopoietic stem cells. The cycle state decides cells' fates, which includes self-renewal, proliferation and differentiation. Proper cell cycle regulation is a pivotal element for the maintenance of hematopoiesis homeostasis. HTm4 is a newly identified specific cell cycle regulator of the hematopoietic cell. Through interacting with KAP-CDK2 complex, it arrests cells in G(0)/G(1) phase. K562 is a human chronic myelogenous leukemia cell; it could be induced to megakaryoblast by phorbol 12-myristate 13 acetate (PMA). Such differentiation must be associated with cell cycle change. To further clarify HTm4's function in hematopoietic cell cycle regulation, K562 cells were treated with PMA. Cell cycle change was analysed using flow cytometric system. And during the induction process gene expression of HTm4 as well as CycleE and CDK2, which are responsible for G(1) to S transition, were analysed using semi-quantitative RT-PCR. The C-terminal domain of HTm4 protein has been shown to be important for HTm4's binding with KAP-CDK2 complex. To determine its impact on HTm4's function, HTm4 and C-terminal truncated HTm4 (HTm4-ct) were transfected into K562 cells using Tet-Off regulation expression system. Their influence on cell cycle was observed. The results showed that PMA induced both expansion and differentiation of K562 cells as measured by cell number count and NBT staining respectively. During PMA treatment, G(0)/G(1) cell proportion and HTm4 expression displayed coordinated change, which suggested that HTm4 might drive K562 cells out of cell cycle but was not involved in the quiescence maintenance. Additionally, transfection of HTm4 caused G(0)/G(1) arrest in K562 cells, while transfection of HTm4-ct did not. It is therefore suggested that the C-terminal domain is important for the function of HTm4 in cell cycle regulation. PMID- 15830104 TI - Effects of various inducers on the expression of P2X7 receptor in human peripheral blood mononuclear cells. AB - Regulation of P2X7 receptor expression is of interest because activation of this receptor by extracellular ATP triggers a wide variety of cell functions in leukocytes. However, its expression and modulation in human peripheral blood mononuclear cells (PBMC) and monocytes remain unclear. RT-PCR was used to detect the constitutive level of P2X7 receptor and the levels upon stimulation with bacteria, bacterial product, mitogen and various cytokines in human PBMC and monocytes. P2X7 receptor mRNA was detected in PBMC and monocytes. P2X7 receptor expression in PBMC was up-regulated by interleukin-2, -4, -6 (IL-2, IL-4, IL-6) tumour necrosis factor-alpha (TNF-alpha), lipopolysaccharide (LPS) and heat inactivated Staphylococcus aureus Cowan strain I (SAC). However, interferon-gamma (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF) and phytohemagglutinin-M (PHA-M) had little effect on the expression of P2X7 receptor. Furthermore, LPS and M-CSF could up-regulate P2X7 receptor expression in monocytes, while IFN-gamma, TNF alpha and GM-CSF had weak effects, but pretreatment with these inducers could not further enhance LPS-stimulated P2X7 receptor expression in monocytes. The results obtained demonstrate that inflammatory stimuli drive P2X7 expression, thus supporting the hypothesis that P2X7 receptor may play a role in the inflammatory responses against bacteria infection, which need further verification. PMID- 15830105 TI - [Promoting effects of serum-free murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells]. AB - To study the effects of serum-free murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of bone marrow endothelial cells, mBMEC-CM was collected and ultrafiltrated by Centriprep-10. The retentate of mBMEC-CM [molecular weight (MW)>10 kDa] and the filtrate of mBMEC-CM (MW<10 kDa) were obtained. The effect of bone marrow conditioned media, their components and exogenous cytokines on the formation of endothelial cell colonies were observed. The effect of bone marrow conditioned media, their components and exogenous cytokines on the proliferation of murine bone marrow endothelial cells were determined by [(3)H]-thymidine incorporation. The method of hybridizing to the Atlas cDNA array was used to determine the expression of cytokine mRNAs in bone marrow endothelial cells. The results obtained are as follows: vWF was expressed in bone marrow endothelial cells. The original mBMEC-CM and MW>10 kDa component of mBMEC-CM promoted the proliferation of bone marrow endothelial cell colonies and increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. The MW<10 kDa component did not affect the production of endothelial cell colonies and did not increase [(3)H]-thymidine incorporation of endothelial cells. Six cytokines (IL-6, IL-11, SCF, GM-CSF, VEGF, bFGF) promoted the proliferation of bone marrow endothelial cell colonies. VEGF, bFGF and SCF increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. According to the results of the Atlas cDNA array, GM-CSF,TGF-beta,BMP-2, bFGF, SCF, endothelin-2, thymosin beta10, MSP-1, connective tissue GF, PDGF-A chain, MIP-2 alpha, PlGF, neutrophil activating protein ENA-78, INF-gamma, IL-1, IL-6, IL-13, IL-11, inhibin-alpha mRNAs were expressed in endothelial cells. These results suggest that murine bone marrow endothelial cell conditioned medium promotes the proliferation of bone marrow endothelial cells. PMID- 15830106 TI - [Mechanism of advanced glycation end products-induced hyperpermeability in endothelial cells]. AB - The purpose of the present study was to investigate the effects of advanced glycation end products (AGEs) modified protein on the permeability of endothelium monolayers and morphological changes of actin cytoskeleton. The roles of receptor for AGEs (RAGE), oxidant stress and the activation of p38 MAPK pathway in this pathological procedure were elucidated. Human umbilical vein endothelial cells (HUVECs)-derived cell line (ECV304) were incubated with AGEs modified human serum albumin (AGE-HSA) in concentrations of 12.5, 25, 50, and 100 microg/ml respectively, for 2, 4, 8, 12 and 24 h. As control, HSA of the same concentration was administered to cells. Then TRITC-albumin was added to evaluate Pa value that reflects the permeability of endothelial monolayer. Furthermore, to visualize the morphological changes of actin cytoskeleton, the treated cells were incubated with rhodamine-phalloidin to stain F-actin. The results showed that the trans endothelial membrane flux of albumin was significantly increased in a concentration- and time-dependent manner upon the stimulation of AGE-HSA, accompanying with actin reorganization. The blockage of AGE and RAGE binding with anti-RAGE IgG and the pharmacological inhibition of NADPH oxidase or p38 MAP kinase greatly attenuated the AGE-induced hyperpermeability response, respectively. These results indicate that RAGE, NADPH oxidase and p38 MAPK are possibly involved in the mediation of AGEs-induced barrier dysfunction and actin cytoskeleton reorganization in endothelial cells. PMID- 15830107 TI - Hydrogen peroxide preconditioning protects PC12 cells against apoptosis induced by oxidative stress. AB - Oxidative stress can induce significant cell death by apoptosis. We explore whether prior exposure to H2O2 (H2O2 preconditioning) protects PC12 cells against the apoptotic consequences of subsequent oxidative damages and what role the ATP sensitive potassium (K(ATP)) channels play in the preconditioning protection. PC12 cells were preconditioned with 90 min exposure to H2O2 at 10 micromol/L, followed by 24-h recovery and subsequent exposures to different concentrations (20, 30, 50 and 100 micromol/L) of H2O2 for 24 h respectively. We used PI staining flow cytometry (FCM) to observe the apoptosis of PC12 cells. It was shown that 24-h exposures to H2O2 at 20, 30, 50 and 100 micromol/L respectively induced substantial cell apoptosis, which was greatly prevented in the preconditioning cells, indicating that H2O2 preconditioning protected PC12 cells against apoptosis induced by H2O2. Administration of pinacidil (10 micromol/L), an K(ATP) channel activator, significantly attenuated the apoptosis of PC12 cells induced by H2O2 at 30 and 50 micromol/L for 24 h respectively. Glybenclamide (10 micromol/L), a K(ATP) channel inhibitor, significantly suppressed or abolished the protective effects caused by the pinacidil but not by H2O2 preconditioning. However, when both H2O2 preconditioning and pinacidil were co-applied, their protection against the apoptosis of PC12 cells was much stronger than that of the individual one of them. These results suggest that H2O2 preconditioning protects PC12 cells against apoptosis and that the activation of K(ATP) channels is not involved in, but synergetically enhances adaptive protection of H2O2 preconditioning. PMID- 15830108 TI - [Extracellular calcium modulates the whole cell potassium currents in Deiters cells isolated from guinea pig cochlea]. AB - To study the modulatory effect of extracellular calcium on the whole cell K(+) currents (I(K)) in isolated Deiters cells, the whole cell K(+) currents were recorded when Deiters cells bathed in normal physiological solutions and calcium free saline, respectively. The electrophysiological characteristics of I(K) currents were then analyzed with the patch clamp technique. Removing extracellular calcium significantly enhanced the amplitude of the I(K) currents, which increased by 70.2% at +50 mV test pulse. The chord conductance, measured at -30 mV test pulse, also significantly increased from (3.31-/+3.08) ns (n=42) in the normal solutions to (10.81-/+6.01) ns (n=42) in the calcium-free solutions, whereas, the zero current potential of the I(K) currents remained unchanged. In calcium-free solutions, the reversal potential of the I(K) currents was shifted to the direction of hyperpolarization, which was very close to the equilibrium K(+) potential based on the Nernst equation. In addition, both the steady state activation curve and the half activation potential, with the averaged value at ( 10.13-/+5.64) mV (n=42), were shifted to the negative. However, the tendency for activation (slope conductances) was the same as that in the normal solutions. Interestingly, both the I-V and the G-V functions deduced from the calcium inhibited K(+) currents in Deiters cells were "S" shape, implying that at least two different kinds of K(+) conductance were involved in this calcium-inhibited K(+) currents. In summary, we hypothesize that there are two mechanisms for this modulation: one is that the I(K) channels in Deiters cells containing a specific calcium sensitive domain, by which extracellular calcium modulates the structure of the K(+) channels and then the I(K) currents; the other is a novel double gated K(+) channel or an ionotropic receptor coupled to K(+) channels or a new subtype of outward K(+) channels. Removing extracellular calcium activates this novel conductance and then modulates the I(K) currents. These results indicate that a decrease in extracellular calcium not only facilitates the efflux of K(+) out of Deiters cells but also accelerates the repolorization by enhancing the I(K) currents, which in turn can effectively buffer the K(+) concentration around the outer hair cells and maintain the resting membrane potential of Deiters cells. PMID- 15830109 TI - Effects of backward masking on the responses of the inferior collicular neurons in the big brown bat, Eptesicus fuscus. AB - Temporal features of sound convey information vital for behaviors as diverse as speech recognition by human and echolocation by bats. However, auditory stimuli presented in temporal proximity might interfere with each other. Although much progress has been made in the description of this phenomenon from psychophysical studies, the neural mechanism responsible for its formation at central auditory structures especially at the inferior colliculus (IC), a midbrain auditory nucleus which practically receives massive bilateral projections from all the major auditory structures in the brainstem, remains unclear. This study was designed to investigate it in vivo by using electrophysiological recording from the inferior collicular neurons of the big brown bat, Eptesicus fuscus. In our results, the responses of 12 (38%, n= 31) neurons to the test sound (leading sound) were obviously inhibited by the masker (lagging sound). The inhibitory effects in these neurons were correlated with the inter-stimulus level difference (SLD) and the inter-stimulus onset asynchrony (SOA) interval. The strength of backward masking increased with the masker intensity increasing, the test sound intensity decreasing and the SOA interval shortening. There were no obvious effects of backward masking on the responses of many other neurons (52%, 16/31), and yet in a part of these neurons, the neural inhibition of responses to the test sound was observed at the special SLD and the special SOA intervals. Moreover, few of the 31 sampled IC neurons (10%, 3/31) displayed facilitating responses to the test sound at the special SLD and the special SOA intervals. These data demonstrate that a lot of IC neurons are involved in the generation of the backward masking of acoustical perception. It is conjectured that the temporal dynamic integration between the leading inhibitory inputs evoked by the masker sound and the excitatory inputs evoked by the test sound might play a key role in shaping the acoustical response characteristics of the IC neurons. PMID- 15830110 TI - Inhibition of thermal hyperalgesia and tactile allodynia by intrathecal administration of gamma-aminobutyric acid transporter-1 inhibitor NO-711 in rats with chronic constriction injury. AB - The present study was undertaken to explore the role of gamma-aminobutyric acid transporters in the neuropathic pain. On the chronic constriction injury (CCI) rats 4 doses (5, 10, 20, 40 microg in group N5, N10, N20, N40, respectively) of specific gamma-aminobutyric acid transporter-1 inhibitor NO-711 or normal saline (in group NS) were intrathecally administered before sciatic nerve ligation (pre treatment) or at the third day after ligation (post-treatment). The paw withdrawl latency (PWL) from a noxious thermal stimulus and paw withdrawl mechanical threshold (PWMT) of von Frey filament was used as measure of thermal hyperalgesia and tactile allodynia respectively. The results demonstrated that post-treatment of NO-711 significantly suppressed thermal hyperalgesia and allodynia in CCI rats (P<0.05, P<0.01), the inhibitory effect lasted for 2 h (N40 group) and 4 h (N20 group) respectively. NO-711 inhibited thermal hyperalgesia induced by CCI in a dose-dependent manner. Intrathecal pretreatment with different doses of NO-711 delayed the occurrence of thermal hyperalgesia, but could not delay the emergence of allodynia induced by CCI. This study indicates that gamma-aminobutyric acid transporter inhibitor has anti-thermal hyperalgesia and anti-tactile allodynia effects in neuropathic rats. PMID- 15830111 TI - Effects of protopine on intracellular calcium and the PKC activity of rat aorta smooth muscle. AB - We have previously shown that the vasodilator effect of protopine (Pro) on rabbit aorta is related to the elevations of cAMP and cGMP. In the present study, the vasodilator mechanisms of Pro were further explored by recording the isotonic contraction of the rat aortic strips, detecting directly the intracellular free Ca(2+) concentration ([Ca(2+)](i)) with Fura-2/AM loaded vascular smooth muscle cells (VSMCs) of rat aorta, and determining the activity of protein kinase C (PKC) in rat aortic tissue with radioactive isotope gamma-32P -ATP-catalyzing assay. By recording the aortic strips contraction induced by noradrenaline (NA) and high potassium (K(+)), Pro shifted nonparallelly the concentration-response curves of NA and high K(+) to right, in which the maximal response was depressed in the presence of Pro (30 and 100 micromol/L), and the values of pD'(2) were 3.70-/+0.25 and 3.97-/+0.15 for NA and high K(+), respectively. In the Fura-2/AM loaded VSMCs, Pro (50 and 100 micromol/L) could not produce any significant change on the resting [Ca(2+)](i), but significantly decreased the [Ca(2+)](i) elevated by NA and high K(+). Pro (30 and 100 micromol/L) had no significant effect on the activity of the cytosolic and membrane PKC in the aortic strips inpretreated by NA. However, in the aortic strips pretreated by NA, the activity of membrane PKC was significantly increased and the activity of cytosolic PKC tended to be decreased by Pro, while the activity of total PKC did not change. These results suggest that Pro seems to promote the translocation of PKC from the cytosol to the membrane in the presence of NA, its vasodilator effect may be the comprehensive result of its decreasing effect on the [Ca(2+)](i) and the increasing effect on cAMP and cGMP, as well as its influence on the PKC. PMID- 15830112 TI - [Stimulation of bone marrow stromal cells conditioned medium on the expansion of mature megakaryocytes and colony forming unit-megakaryocyte in vitro]. AB - In this study the effects of bone marrow stromal cells conditioned medium on the expansion of mature megakaryocytes and colony forming unit-megakaryocyte (CFU Meg) in vitro were investigated. The serum-free bone marrow fibroblast conditioned medium (F-CM), bone marrow endothelial cell conditioned medium (E-CM) and bone marrow macrophage conditioned medium (M-CM) were collected and ultrafiltrated by using Centriprep-10. F-CM, E-CM, M-CM, the retentate (>10 kDa F CM, >10 kDa E-CM and >10 kDa M-CM) contained substances whose molecular weight was more than 10 kDa and the filtrate (<10 kDa F-CM, <10 kDa E-CM and <10 kDa M CM) contained substances whose molecular weight was less than 10 kDa were added in liquid culture system respectively. The results showed that F-CM, >10 kDa F-CM and E-CM, >10 kDa E-CM significantly promoted the expansion of mature megakaryocytes in liquid culture system, the percentage of mature megakaryocytes compared with 0 h control were (287.33-/+16.77)%, (236.67-/+39.72)%, (141.21 /+17.47)% and (179.03-/+30.98)%. But <10 kDa F-CM, <10 kDa E-CM, M-CM, >10 kDa M CM and <10 kDa M-CM had no positive effects on the expansion of mature megakaryocytes. The effects of F-CM, E-CM or M-CM on the expansion of CFU-Meg were also investigated. The results indicated that F-CM and E-CM promoted the expansion of CFU-Meg in liquid culture system. M-CM had no positive effect on the expansion of CFU-Meg. the percentage of CFU-Meg compared with 0 h control were (168.18-/+30.24)%, (215.17-/+17.4)% and (85.0-/+7.0)%. The results of reverse transcription-polymerase chain reaction (RT-PCR ) showed that transforming growth factor -beta1 (TGF-beta1) mRNA was expressed in bone marrow endothelial cells and was not expressed in bone marrow fibroblasts. Thrombopoietin (TPO) mRNA was expressed in bone marrow fibroblasts and was not expressed in bone marrow endothelial cells. These results suggest that F-CM, >10kDa F-CM and E-CM, >10kDa E-CM significantly promoted the expansion of mature megakaryocytes and CFU-Meg in liquid culture system. The effect of F-CM on the expansion of mature megakaryocytes is better than that of E-CM. PMID- 15830113 TI - [Effect of leptin on growth hormone secretion and apoptosis of GH3 cells]. AB - In order to investigate the effect of leptin on the secretion of rat pituitary adenoma GH3 cell and its mechanisms, we observed the effect of leptin on the growth hormone secretion, proliferation and apoptosis of GH3 cells. The results indicated that leptin at 1, 10, and 100 nmol/L could inhibit the basal growth hormone secretion of GH3 cells in a dose dependent manner (P<0.05). Short-term treatment of leptin (10 nmol/L) for 30 min, 1 and 3 h did not affect basal GH secretion. However, treatment of the GH3 cells with leptin (10 nmol/L) for 1 d or longer resulted in an inhibition of GH secretion (P<0.05). We used MTT method and flow cytometery (FCM) to study the effect of leptin on the proliferation and apoptosis of GH3 cells. We found that leptin inhibited proliferation of GH3 cells with a dose-dependent manner. And leptin reduced the proportion of cells in S phase, increased the proportion of cells in G1, and increased the proportion of GH3 cells in 2 and 4 phase. These results demonstrate that leptin inhibits the basal GH secretion of GH3 cells, which may be due to the inhibition of DNA synthesis and advanced apoptosis of GH3 cells. PMID- 15830114 TI - [Endothelial cells and cardiac explants in three-dimensional culture system]. AB - To observe the morphological features of endothelial cells and cardiac explants cultured in two- or three-dimensional culture systems, several three-dimensional collagen type I culture systems, such as the in gel, on gel, sandwich model, and the microscope slide model, were used to examine the growth patterns of the cells and explants from heart by using immunofluorescence staining and microscopic observation in the presence or absence of vascular endothelial growth factor (VEGF). In two-dimensional cultures the primary cardiac endothelial cells arrayed into a cobblestone-like structure. When cultured in three-dimensional matrix, the cells were elongated and migrated into the gel, with a phenotype similar to that in the process of angiogenesis and vasculogenesis in vivo. VEGF promoted the process of the endothelial cells transforming into tube-like structure. Cardiac explants grew well in the collagen gel. Adjacent explants were connected to each other by the migrating cells with the occurrence of autorhythmic beating of the explants. Thin-layer collagen gel, microscope slide chamber and aorta-strip model were also tested and proved to be good tools for vasculogenesis or angiogenesis studies. Three-dimensional culture systems enable the endothelial cells to proliferate, migrate, and anchor to three-dimensional vascular structures, showing advantages for observing the feature of angiogenesis. Different three dimensional culture models may be used for variable research purposes. PMID- 15830115 TI - The history and evolution of pessaries for pelvic organ prolapse. AB - The use of pessaries for the treatment of genital prolapse dates back prior to the days of Hippocrates and their use has been documented in early Egyptian papyruses. Throughout the centuries remedies such as honey, hot oil, wine and fumes have been used as treatment. Mechanical methods included succussion and leg binding. Pomegranates were also common remedies. In the middle ages, linen and cotton wool soaked in many different potions were used. As new materials were discovered, pessaries evolved and began to resemble those used today. Cork and brass were soon replaced with rubber. Modern day pessaries are made of non reactive silicone and come in various designs and sizes to suit each individual. Pessaries can be used as an interim measure for women who wish to complete childbearing or women awaiting surgery. It can also be used as a permanent measure for women who are unsuitable for surgery. It remains to be established whether the use of modern pessaries over prolonged periods of time can prevent progression of or even cure, prolapse. PMID- 15830116 TI - Visualization of the endopelvic fascia by transrectal three-dimensional ultrasound. AB - The aim of our pilot study was to explore the feasibility of visualizing the endopelvic fascia by transrectal three-dimensional (3D) ultrasound. Transrectal 3D ultrasound was performed in 12 nulliparous women and 11 women with a history of vaginal delivery. A 6-10 MHz volume probe was used to examine the suburethral anterior vaginal wall. In all women, an echogenic layer was identified at an average of 3-5 mm from the vaginal surface. This echogenic layer was found to be contiguous to the lateral pelvic sidewall and uninterrupted in 10 of 12 nulliparous women, whereas gaps in this layer were identified in all 11 parous women. We hypothesize that this echogenic layer may represent the suburethral component of the endopelvic fascia. Depending on the number and localization of the interruptions in this echogenic layer, the mechanical support of the pelvic floor seems to be weakened corresponding to a higher incidence of descensus of the anterior vaginal wall, which frequently was associated with urinary incontinence. PMID- 15830117 TI - How to appraise a prognostic study. AB - Prognostic studies are studies that examine selected predictive variables or risk factors and assess their influence on the outcome of a disease. They allow clinicians to understand better the natural history of a disease, guide clinical decision-making by facilitating the selection of appropriate treatment options, and allow more accurate prediction of disease outcomes. Appraising prognostic studies involves determining the internal validity of the study design and evaluating the influence of systemic errors or bias. In studies examining multiple prognostic variables, care must be taken to minimize the confounding influence each variable would have on the other parameters. Evaluating the results of appropriate statistical analysis enables conclusions to be made that may influence clinical practice. Care must be taken to ensure that the conditions under which the prognostic study were conducted resemble circumstances in the local institution so as to allow the conclusions to be applied to local practices. PMID- 15830118 TI - A great Russian surgeon: Boris Dmitrievich Savchuk (1933-2004). PMID- 15830120 TI - Highlights in surgery through outstanding ISS/SIC surgeons: Boris Vasilyevich Petrovsky. PMID- 15830121 TI - Effects of formate on fermentative hydrogen production by Enterobacter aerogenes. AB - This paper describes the effects of formate on fermentative hydrogen production by Enterobacter aerogenes by way of batch culture. When 20 mM formate was added to pH 6.3 and pH 5.8 E. aerogenes glucose cultures (formate culture) at the beginning of cultivation, hydrogen evolution through both glucose consumption and decomposition of the extrinsic formate occurred together, while hydrogen evolution occurred only through glucose consumption in the control cultures. The hydrogen evolution rates in the formate cultures were faster than in the control cultures, although cell growth and glucose consumption rates in the formate cultures were slower than the control cultures'. The decomposition rate of the extrinsic formate in the pH 5.8 formate culture was faster than in the pH 6.3 formate culture. The hydrogen yield from glucose in the pH 6.3 formate culture increased due to the increasing amount of the nicotinamide adenine dinucleotide for hydrogen production. PMID- 15830122 TI - Association of nucleotide variations in the apolipoprotein B48 receptor gene (APOB48R) with hypercholesterolemia. AB - Factors predisposing to the phenotypic features of high total cholesterol (T-Cho) in human plasma have not been clearly defined. Here we report an association between two variations in the apolipoprotein B48 receptor gene (APOB48R) and plasma T-Cho levels among 352 adult individuals in Japan. By analyzing phenotypic associations between age- and gender-adjusted levels of plasma T-Cho, low-density lipoprotein (LDL) cholesterol (LDL-C), and high-density lipoprotein (HDL) cholesterol (HDL-C), we detected a significant correlation between genotypes of the A419P variation and adjusted T-Cho levels. Among homozygous G-allele carriers (n=265), heterozygous carriers (n=78), and homozygous minor C-allele carriers (n=9), T-Cho levels were 2.43+/-0.21 mg/cm(3), 2.48+/-0.24 mg/cm(3), and 2.63+/ 0.21 mg/cm(3), respectively, indicating a codominant T-Cho-elevating effect of the minor C-allele (r=0.15, P=0.007). A similar effect was detected for c.934 960/del (r=0.13, P=0.015). Linkage disequilibrium (LD) analysis detected significant LD among eight variant sites that included neighboring loci. Our results indicate that variations in APOB48R and nearby genes are among the many factors involved in hypercholesterolemia. The etiological studies should now include consideration of this novel aspect of the mechanism(s) leading to hypercholesterolemic disease. PMID- 15830123 TI - Inhibition of phospholipase A2 activity reduces membrane fluidity in rat hippocampus. AB - Phospholipase A2 (PLA2) is a family of key enzymes in membrane phospholipid metabolism. In rats, the inhibition of PLA2 activity in the hippocampus was found to impair memory formation. Because memory function is largely dependent on the fluidity of brain membranes, we performed the present study to investigate the effects of in vivo PLA2 inhibition (with PACOCF3) on the fluidity of hippocampal membranes from rats trained in a learning task. Hippocampal tissue from rats injected with 100 microM PACOCF3 showed reduced membrane fluidity as compared to vehicle (p < 0.01), and the reduction of membrane fluidity was highly correlated with PLA2 inhibition (r = .76, p < 0.03). This finding is of interest because reduction of brain membrane fluidity impairs memory formation and both decreased PLA2 activity and reduced membrane fluidity have been reported in the brain from patients with Alzheimer's disease. PMID- 15830124 TI - Detailed expression analysis of selected genes of the aldehyde dehydrogenase (ALDH) gene superfamily in Arabidopsis thaliana. AB - Aldehyde dehydrogenase (ALDH) genes have been identified in almost all organisms from prokaryotes to eukaryotes, but particularly in plants knowledge is very limited with respect to their function. The data presented here are a contribution towards a functional analysis of selected Arabidopsis ALDH genes by using expression profiles in wild types and mutants. The Arabidopsis thaliana genome contains 14 genes which represent 9 families. To gain insight into the possible roles of aldehyde dehydrogenases from Arabidopsis, the expression patterns of five selected ALDH genes were analyzed under defined physiological conditions. Three genes (ALDH3I1, 3H1 and ALDH7B4) that belong to two different families are differentially activated by dehydration, high salinity and ABA in a tissue-specific manner. The other two genes (ALDH3F1 and ALDH22A1) are constitutively expressed at a low level. Transcript analysis of ALDH3I1 and ALDH7B4 in Arabidopsis mutants suggests that stress responses are differentially controlled by the phytohormone ABA as well as by pathways that affect sugar metabolism and fatty acid composition of membrane lipids. Our results indicate that the stress-associated ALDH genes participate in several pathways and that their regulation involves diverged signal transduction pathways. PMID- 15830125 TI - Protein import pathways in 'complex' chloroplasts derived from secondary endosymbiosis involving a red algal ancestor. AB - Heterokont algae such as diatoms and the raphidophyte Heterosigma akashiwo and peridinin-containing dinoflagellates such as Heterocapsa triquetra originally acquired their chloroplasts via secondary endosymbiosis involving a red algal endosymbiont and a eukaryote host, resulting in 'complex' chloroplasts surrounded by four and three membranes, respectively. The precursors of both heterokont and dinoflagellate chloroplast-targeted proteins are first inserted into the ER with removal of an N-terminal signal peptide, but how they traverse the remaining membranes is unclear. Using a nuclear-encoded thylakoid lumen protein, PsbO, from the heterokont alga Heterosigma akashiwo, the dinoflagellate Heterocapsa triquetra and the red alga Porphyra yezoensis we show that precursors without the ER signal peptide can be imported into pea chloroplasts. In the case of the H. triquetra and Porphyra PsbO, the precursors were processed to their predicted mature size and localized within the thylakoid lumen, using the Sec-dependent pathway. We report for the first time a stromal processing peptidase (SPP) activity from an alga of the red lineage. The enzyme processes the Heterosigma PsbO precursor at a single site and appears to have different substrate and reaction specificities from the plant SPP. In spite of the fact that we could not find convincing homologs of the plant chloroplast import machinery in heterokont (diatom) and red algal genomes, it is clear that these three very different lines of algae use similar mechanisms to import chloroplast precursors. PMID- 15830126 TI - Arabidopsis cue mutants with defective plastids are impaired primarily in the photocontrol of expression of photosynthesis-associated nuclear genes. AB - Plant photoreceptors detect light cues and initiate responses ranging from chloroplast differentiation to the control of morphogenesis and flowering. The photocontrol of photosynthesis-related nuclear genes appears closely related to 'retrograde plastid signals' by which the status of the organelle controls the expression of nuclear genes. However, what specific role, if any, plastid originated signals play in light responses is poorly understood: it has in the past been proposed that plastid signals play a role in all responses to 'high fluence' far-red light perceived by the light-labile phytochrome A, irrespective of whether they involve photosynthesis-related genes. To explore this further, we have re-examined the phenotype of three cue (cab-underexpressed) Arabidopsis mutants, defective in chloroplast development. The mutants have underdeveloped etioplasts, with increasing impairments in cue6, cue8 and cue3. The mutants show only small defects in photocontrol of hypocotyl elongation and cotyledon opening under prolonged far-red or red light, and normal photocontrol under blue. On the other hand, the expression of photosynthesis-associated nuclear genes is much more impaired in the mutants in the dark and following red or far-red light short treatments or continuous light, than that of those phytochrome-dependent genes tested which are not associated with photosynthesis. Furthermore, red/far-red photoreversible responses involving photosynthesis-related genes (induction of Lhcb1-cab promoter activity, and photoreversible extent of greening) mediated by phytochrome B and other photo-stable phytochromes, both show a reduction in the cue mutants, which correlates with the etioplast defect. Our evidence demonstrates that plastid-derived signals need to be operational in order for the phytochrome control of photosynthetic nuclear genes to occur. PMID- 15830127 TI - A Kunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts anti metabolic effect on podborer (Helicoverpa armigera) larvae. AB - Chickpea (Cicer arietinum L.) seeds contain Bowman-Birk proteinase inhibitors, which are ineffective against the digestive proteinases of larvae of the insect pest Helicoverpa armigera. We have identified and purified a low expressing proteinase inhibitor (PI), distinct from the Bowman-Birk Inhibitors and active against H. armigera gut proteinases (HGP), from chickpea seeds. N-terminal sequencing of this HGP inhibitor revealed a sequence similar to reported pea (Pisum sativum) and chickpea alpha-l-fucosidases and also homologous to legume Kunitz inhibitors. The identity was confirmed by matrix assisted laser desorption ionization - time of flight analysis of tryptic peptides and isolation of DNA sequence coding for the mature protein. Available sequence data showed that this protein forms a distinct phylogenetic cluster with Kunitz inhibitors from Glycine max, Medicago truncatula, P. sativum and Canavalia lineata. The isolated coding sequence was cloned into a yeast expression vector and produced as a recombinant protein in Pichia pastoris. alpha-l-fucosidase activity was not detectable in purified or recombinant protein, by solution assays. The recombinant protein did not inhibit chymotrypsin or subtilisin activity but did exhibit stoichiometric inhibition of trypsin, comparable to soybean Kunitz trypsin inhibitor. The recombinant protein exhibited higher inhibition of total HGP activity as compared to soybean kunitz inhibitor, even though it preferentially inhibited HGP trypsins. H. armigera larvae fed on inhibitor-incorporated artificial diet showed significant reduction in average larval weight after 18 days of feeding demonstrating potent antimetabolic activity. The over-expression of this gene in chickpea could act as an endogenous source of resistance to H. armigera. PMID- 15830129 TI - Gene expression profile in response to Xanthomonas axonopodis pv. manihotis infection in cassava using a cDNA microarray. AB - A cassava cDNA microarray based on a large cassava EST database was constructed and used to study the incompatible interaction between cassava and Xanthomonas axonopodis pv. manihotis (Xam) strain CIO151. For microarray construction, 5700 clones from the cassava unigene set were amplified by polymerase chain reaction (PCR) and printed on glass slides. Microarray hybridization was performed using cDNA from cassava plants (resistant variety MBra685) collected at 12, 24, 48 h and 7 and 15 days post-infection as treatment and cDNA from mock-inoculated plants as control. A total of 199 genes were found to be differentially expressed (126 up-regulated and 73 down-regulated). A greater proportion of differentially expressed genes was observed at 7 days after inoculation. Expression profiling and cluster analyses indicate that, in response to inoculation with Xam, cassava induces dozens of genes, including principally those involved in oxidative burst, protein degradation and pathogenesis-related (PR) genes. In contrast, genes encoding proteins that are involved in photosynthesis and metabolism were down regulated. In addition, various other genes encoding proteins with unknown function or showing no similarity to other proteins were also induced. Quantitative real time PCR experiments confirmed the reliability of our microarray data. In addition we showed that some genes are induced more rapidly in the resistant than in the susceptible cultivar. PMID- 15830130 TI - Gene regulation in planta by plant-derived engineered zinc finger protein transcription factors. AB - The ability to modify plant traits is of great commercial potential in agricultural biotechnology. To this end we have engineered plant-based zinc finger protein transcription factors (ZFP TFs) that minimize the use of non-plant DNA sequences. This novel architecture supports the use of tandem arrays of zinc finger DNA recognition domains such that the ZFP TF binds a contiguous DNA target site - thus emulating the design of ZFP TFs described previously for mammalian gene regulation. We show that this plant-based ZFP TF architecture supports high affinity DNA binding while allowing the specificity of the DNA-protein interaction to be determined by the amino acid sequences of the recognition helices. This plant-based backbone thus supports the use of previously characterized DNA recognition helices originally identified in a mammalian ZFP context without using mammalian DNA sequences. Moreover, we show that plant-based ZFP TFs employing this new architecture can up-regulate endogenous ADH activity by > 20-fold in transgenic Arabidopsis. Thus plant-based ZFP TFs are shown to be potent regulators of gene expression in vivo. PMID- 15830131 TI - The dsdA gene from Escherichia coli provides a novel selectable marker for plant transformation. AB - Plants are sensitive to D-serine, but functional expression of the dsdA gene, encoding D-serine ammonia lyase, from Escherichia coli can alleviate this toxicity. Plants, in contrast to many other organisms, lack the common pathway for oxidative deamination of D-amino acids. This difference in metabolism has major consequences for plant responses to D-amino acids, since several D-amino acids are toxic to plants even at relatively low concentrations. Therefore, introducing an enzyme specific for a phytotoxic D-amino acid should generate a selectable characteristic that can be screened. Here we present the use of the dsdA gene as a selectable marker for transformation of Arabidopsis. D-serine ammonia lyase catalyses the deamination of D-serine into pyruvate, water and ammonium. dsdA transgenic seedlings can be clearly distinguished from wild type, having an unambiguous phenotype immediately following germination when selected on D-serine containing medium. The dsdA marker allows flexibility in application of the selective agent: it can be applied in sterile plates, in foliar sprays or in liquid culture. Selection with D-serine resistance was compared with selection based on kanamycin resistance, and was found to generate similar transformation frequencies but also to be more unambiguous, more rapid and more versatile with respect to the way the selective agent can be supplied. PMID- 15830132 TI - Site-specific recombination in Arabidopsis plants promoted by the Integrase protein of coliphage HK022. AB - The gene encoding the wild type Integrase protein of coliphage HK022 was integrated chromosomally and expressed in Arabidopsis thaliana plants. Double transgenic plants cloned with the int gene as well as with a T-DNA fragment carrying the proper att sites in a tandem orientation showed that Int catalyzed a site-specific integration reaction (attP x attB) as well as a site-specific excision reaction (attL x attR). The reactions took place without the need to provide any of the accessory proteins that are required by Int in the bacterial host. When expressed in tobacco plants a GFP-Int fusion exhibits a predominant nuclear localization. PMID- 15830133 TI - Evaluation of five ab initio gene prediction programs for the discovery of maize genes. AB - Five ab initio programs (FGENESH, GeneMark.hmm, GENSCAN, GlimmerR and Grail) were evaluated for their accuracy in predicting maize genes. Two of these programs, GeneMark.hmm and GENSCAN had been trained for maize; FGENESH had been trained for monocots (including maize), and the others had been trained for rice or Arabidopsis. Initial evaluations were conducted using eight maize genes (gl8a, pdc2, pdc3, rf2c, rf2d, rf2e1, rth1, and rth3) of which the sequences were not released to the public prior to conducting this evaluation. The significant advantage of this data set for this evaluation is that these genes could not have been included in the training sets of the prediction programs. FGENESH yielded the most accurate and GeneMark.hmm the second most accurate predictions. The five programs were used in conjunction with RT-PCR to identify and establish the structures of two new genes in the a1-sh2 interval of the maize genome. FGENESH, GeneMark.hmm and GENSCAN were tested on a larger data set consisting of maize assembled genomic islands (MAGIs) that had been aligned to ESTs. FGENESH, GeneMark.hmm and GENSCAN correctly predicted gene models in 773, 625, and 371 MAGIs, respectively, out of the 1353 MAGIs that comprise data set 2. PMID- 15830128 TI - Development of a citrus genome-wide EST collection and cDNA microarray as resources for genomic studies. AB - A functional genomics project has been initiated to approach the molecular characterization of the main biological and agronomical traits of citrus. As a key part of this project, a citrus EST collection has been generated from 25 cDNA libraries covering different tissues, developmental stages and stress conditions. The collection includes a total of 22,635 high-quality ESTs, grouped in 11,836 putative unigenes, which represent at least one third of the estimated number of genes in the citrus genome. Functional annotation of unigenes which have Arabidopsis orthologues (68% of all unigenes) revealed gene representation in every major functional category, suggesting that a genome-wide EST collection was obtained. A Citrus clementina Hort. ex Tan. cv. Clemenules genomic library, that will contribute to further characterization of relevant genes, has also been constructed. To initiate the analysis of citrus transcriptome, we have developed a cDNA microarray containing 12,672 probes corresponding to 6875 putative unigenes of the collection. Technical characterization of the microarray showed high intra- and inter-array reproducibility, as well as a good range of sensitivity. We have also validated gene expression data achieved with this microarray through an independent technique such as RNA gel blot analysis. PMID- 15830134 TI - Gemcitabine monotherapy as salvage chemotherapy in heavily pretreated metastatic breast cancer. AB - PURPOSES: We evaluated the efficacy and tolerability of gemcitabine monotherapy in heavily pretreated, breast cancer patients as salvage chemotherapy. METHODS: A weekly infusion of gemcitabine at 850 mg/m2 for 30 min, for 3 of every 4 weeks, was introduced in advanced breast cancer patients who had failed previous doxorubicin and taxane based chemotherapy. There was no dose modification, and the treatment was delayed until the leukopenia was recovered with G-CSF support. The efficacy was evaluated every three cycles and the treatment was continued until either disease progression or 12 cycles. RESULTS: Of 41 enrolled patients, 38 were evaluable with a median age of 47. Total 178 cycles of gemcitabine was administered and the relative dose intensity was 89. The toxicity was mild with 12 of grade III neutropenia and 14 of grade III/IV thrombocytopenia without clinical symptoms. The response rate was 20 (8/38), comprising two complete and six partial responses. The median response duration and overall survival were 9 (2-25) and 11 months, respectively. The overall survival of 12 months was better in the third line patients than the 7 months in the fourth line treatment group. CONCLUSION: Gemcitabine monotherapy is effective and safe as salvage treatment in heavily pretreated, breast cancer patients. PMID- 15830135 TI - Women with multiple primary breast cancers diagnosed within a five year period, 1994-1998. AB - INTRODUCTION: Women diagnosed with a primary breast cancer are at higher risk for a second primary. Few studies have focused on a comparison of women with single breast primary cancers and women with multiple primary breast cancers. The 1994 1998 NAACCR dataset aggregated from high quality registries representing more than one-third of the US population provides a unique opportunity to examine the incidence of multiple primary breast cancers in a large population. MATERIALS AND METHODS: Using this multi-registry dataset, we describe the incidence pattern of malignant synchronous (within 2 months) and short-term metachronous (from 3 to 60 months) multiple primaries and single primary breast cancers by demographic and tumor characteristics. RESULTS: Synchronous multiple primary tumors were more similar in age, stage, and tumor grade to single breast tumors than they were to short-term metachronous tumors. The short-term metachronous tumors did not resemble either the synchronous tumors or the single primaries. DISCUSSION: These findings may indicate that while synchronous multiple primaries may have treatment implications different from single primaries, their etiology may be similar to single breast primaries. Further, they may actually be multi-centric single primaries. The two-month interval between multiple primaries is arbitrary and may not distinguish between the synchronous tumors and those occurring within 12 months of the index tumor. The rules for defining and counting breast primaries have implications for interpretation of incidence rates and temporal trends. These data also suggest the need for standard definitions for multiple primary breast tumors among clinicians, pathologists, and surveillance epidemiologists. PMID- 15830136 TI - Alteration of the effects of cancer therapy agents on breast cancer cells by the herbal medicine black cohosh. AB - Recent studies have revealed that many, perhaps most, patients receiving cancer therapy are simultaneously self-medicating with one or several complementary and alternative medicines, often without discussing the use of these agents with their physicians. The effects of these agents on the efficacy and toxicity of standard anticancer therapy have not been studied. The experiments described in this report used a well characterized mouse breast cancer cell line to ask whether commercially available extracts of black cohosh, an herb widely used by breast cancer patients, altered the response of cancer cells to radiation and to four drugs commonly used in cancer therapy. The black cohosh extracts increased the cytotoxicity of doxorubicin and docetaxel and decreased the cytotoxicity of cisplatin, but did not alter the effects of radiation or 4 hydroperoxycyclophosphamide (4-HC), an analog of cyclophosphamide which is active in cell culture. These data sound a warning that the herbal medicines being used by patients undergoing cancer therapy can have effects on cancer cells that alter their response to the agents commonly used to treat breast cancer. PMID- 15830137 TI - Expression of BP1, a novel homeobox gene, correlates with breast cancer progression and invasion. AB - BACKGROUND: Our previous studies revealed that the mRNA encoded by BP1, a member of the homeobox gene superfamily of transcription factors, was expressed in leukemia and infiltrating breast ductal carcinoma (IDC). This study investigated the immunohistochemical profile of BP1, to determine whether the expression of BP1 protein correlated with breast tumor progression and invasion and whether BP1 was co-localized with erbB2. DESIGN: Paraffin sections from normal reduction mammoplasties (n = 34) and a variety of in situ and invasive breast cancers (n = 270) were either singly immunostained for BP1, or doubly immunostained for BP1 plus either erbB2 or Ki-67. RESULTS: The prevalence of BP1 positive cells and the intensity of BP1 immunoreactivity increased with the extent of ductal proliferation and carcinogenesis. BP1 expression was barely detectable in normal reduction mammoplasties compared to distinct staining in 21, 46, and 81% of hyperplastic, in situ, and infiltrating lesions, respectively. In cases with co existing normal, hyperplastic, in situ, and invasive lesions, the tumor cells of the invasive lesions consistently showed the highest frequency and the highest intensity of BP1 immunostaining, followed by in situ tumor cells. Double immunostaining revealed that BP1 co-localized with a subset of erbB2 positive cells in all 15 in situ and IDC tumors examined, and that BP1 positive cells had a substantially higher proliferation rate than morphologically similar cells without BP1 expression. CONCLUSION: These findings suggest that BP1 is an important upstream factor in an oncogenic pathway, and that expression of BP1 may reliably reflect or directly contribute to tumor progression and/or invasion. PMID- 15830138 TI - Biomarker profile and genetic abnormalities in lobular carcinoma in situ. AB - The predisposition of patients with lobular carcinoma in situ (LCIS) to develop invasive breast cancer (IBC) is well known. However, relatively little is known about the biologic characteristics, which may be involved in the development and progression of LCIS. This study evaluated 59 cases of LCIS (29 pure, 30 with synchronous IBC) for five biomarkers known to be important in IBC (ER, PgR, c erbB-2, p53 and Ki-67 proliferation rate) by immunohistochemistry. A comprehensive analysis of loss-of-heterozygosity (LOH) was performed in 12 cases (10 pure, 2 with synchronous IBC) at 15 genetic loci on 9 chromosomes. LCIS demonstrated a low grade/favorable biophenotype that was not significantly different in cases with and without synchronous IBC (ER 98%, PgR 84%, c-erbB-24%, p53 19% and proliferation rate 2%). LOH was present in 80% of pure LCIS and the highest rates of LOH were at loci on 9p (30%), 16q (63%), 17p (33%) and 17q (50%). The clustering of LOH at these four foci suggests that inactivated tumor suppressor genes in these regions may be particularly important. LOH was present in both cases of LCIS with synchronous IBC and the LOH phenotype was shared by LCIS and IBC. Our findings suggest that five known prognostic factors in IBC do not have prognostic utility in LCIS. Multiple genetic mechanisms may be involved in the development of LCIS. PMID- 15830139 TI - Influences of apolipoprotein E polymorphism on the risk for breast cancer and HER2/neu status in Taiwan. AB - Apolipoprotein E (APOE) polymorphism plays an important role in lipid metabolism. Preliminary evidence suggests that APOE genotype appears to be a risk factor for not only cardiovascular disease, but also Alzheimer's disease and cancer. We screened the APOE genotype in 290 breast cancer patients and 232 non-cancer controls and determined the relationship between APOE gene polymorphism and breast cancer in Taiwan. We found risk for breast cancer was associated with the APOE genotype (xi(2) = 8.652, p = 0.013). Carriers of the epsilon4 allele were more common in breast cancer cases than carriers of epsilon3 allele (p = 0.004, OR = 1.786, 95% CI: 1.197-2.664). In addition, the epsilon4 allele is also associated with HER2/neu negative status in breast cancer patients (p = 0.006, OR = 0.277, 95% CI: 0.111-0.693). No significant associations between APOE genotype and tumor grade, TN classification, progesterone receptor, estrogen receptor, lymphatic invasion, or recurrence of breast cancer were in evidence. These results suggest that the APOE epsilon4 allele may be a risk factor for breast cancer and correlates with HER2/neu negative status. PMID- 15830140 TI - Delayed versus immediate exercises following surgery for breast cancer: a systematic review. AB - BACKGROUND: Seroma formation, wound healing and fluid drainage are a concern for both surgeons and patients. Excessive fluid production can result in seroma formation, and inadequate drainage of seromas is known to cause infection, pain, discomfort and longer periods of hospitalisation. Postoperative exercises given to maintain movement of the arm are believed to increase the amount of fluid production following surgery. This review aimed to determine whether a program of delayed exercises reduces the risk of seroma formation, fluid loss and hospital stay, without loss of arm movement. METHOD: A systematic review. RCTs of early versus delayed shoulder mobilisation after surgery in females with breast cancer were included in the review. Outcomes. One or more measurements of shoulder range of motion, wound complications, fluid drainage volumes and incidence of seroma formation. Design. Randomised controlled trials, control group of delayed exercise/mobilisation. Validity assessment was carried out using a data extraction form based on the CONSORT statement. Study characteristics recorded include sample size, intervention, control, period of exercise delay, surgical procedure and conclusions drawn. Data synthesis was carried out using random effects and weighted mean differences to test for heterogeneity and combined effects. RESULTS: 12 RCTs were included in the review of which 6 were included for meta-analysis. Delaying exercises significantly decreases seroma formation (OR=0.4; 95%CI 0.2-0.5; p=0.00001). No significant differences were found for drainage volume or hospital stay. CONCLUSION: Current evidence from RCTs supports the use of a delayed program of arm exercises to reduce seroma formation. Clinical and statistical inconsistencies between studies did not allow any conclusions to be drawn regarding the effects of delayed exercises on fluid drainage, hospital stay and immediate or long term ability to move the arm. PMID- 15830141 TI - Dual role of AP-2gamma in ErbB-2-induced mammary tumorigenesis. AB - A causative role of the membrane-bound tyrosine kinase ErbB-2 in breast tumorigenesis has been well established. MMTV/neu transgenic mice which overexpress ErbB-2 consistently develop mammary carcinomas with a high incidence. In human breast cancer, ErbB-2 is overexpressed in 25-30 of all cases and is representing a clinical marker of a poor prognosis. Besides to gene amplification, ErbB-2 overexpression has been attributed to transcription factors of the AP-2 family which were shown to control the ErbB-2 gene promoter in cell culture studies. Particularly AP-2alpha and gamma are often coexpressed in ErbB-2 positive breast carcinomas. However, LTRgamma transgenic mice which overexpress AP-2gamma in their mammary epithelium display only a very weak upregulation of the erbB-2 gene and do not develop mammary carcinoma. These findings therefore raise the possibility of functional cooperativity between both genes in breast cancer. To experimentally address the impact of AP-2gammaon ErbB-2-induced breast carcinogenesis we crossed MMTV/neu transgenic mice with LTRgamma transgenic mice and monitored tumor development in bitransgenic female progeny. AP-2gamma overexpression negatively influenced tumor incidence, as reflected by a reduced tumor number and a prolonged tumor latency. Histological analysis revealed three major types of tumors corresponding to different stages of tumor progression. Interestingly, an increased proportion of advanced stage carcinomas was observed in bitransgenic mice. Moreover, the AP-2gamma transgene differentially affected proliferation rates between the different progression stages: proliferation was enhanced at early stages but reduced in advanced stages in comparison to control tumors. Therefore, AP-2gamma while reducing the incidence of mammary tumors is promoting tumor progression. PMID- 15830143 TI - Validation of the Functional Assessment of Cancer Therapy-Breast Symptom Index (FBSI). AB - We assessed the reliability, validity, and responsiveness to change of the 6-item Functional Assessment of Cancer Therapy-Breast Symptom Index (FBSI) in a sample of 615 metastatic breast cancer patients. The FBSI is a brief, clinically relevant, and psychometrically sound instrument that can be used to measure symptoms in patients with breast cancer. PMID- 15830142 TI - Development of a prolactin receptor-targeting fusion toxin using a prolactin antagonist and a recombinant form of Pseudomonas exotoxin A. AB - Human prolactin (hPRL) promotes the proliferation and differentiation of mammary epithelial cells during mammary gland development and has been linked to breast tumor development. The receptor for hPRL (hPRL-R) is elevated in a majority of human breast tumors, suggesting the overexpression of hPRL-R makes cancer cells highly sensitive to the mitogenic and anti-apoptotic activity of hPRL. These findings provide the rationale for the development of hPRL-R targeting breast cancer therapeutics. Previously, an hPRL-R antagonist, G129R, was developed that competitively binds to the hPRL-R resulting in growth inhibition and the induction of apoptosis in certain types of breast cancer cells. To further increase the potency of G129R, we fused G129R to a truncated form of Pseudomonas exotoxin A (PE(40)) that lacks the cell recognition domain of the toxin but retains the domains necessary for PE(40)_ to translocate into the cytosol and inhibit protein synthesis. We postulated that the fusion of G129R with PE(40) KDEL would (1) deliver the recombinant toxin to breast cancer cells where hPRL-R is overexpressed; (2) block hPRL signaling via its G129R moiety; and (3) inhibit protein synthesis via its PE(40)-KDEL moiety. We demonstrate that the fusion toxin can competitively bind to hPRL-Rs on T-47D human breast cancer cells and inhibit STAT5 phosphorylation induced by hPRL. In addition, we show that G129R PE(40)-KDEL is selectively cytotoxic to breast cancer cell lines expressing the hPRL-R and that cell death is associated with the inhibition of protein synthesis and does not involve caspase mediated apoptosis. PMID- 15830144 TI - Accuracy of breast screening among women with and without a family history of breast and/or ovarian cancer. AB - OBJECTIVES: To compare interval cancer rates, sensitivity and specificity of breast cancer screening between women with moderate or strong family history and women without a family history. METHODS: From 1996 to 1997, 115,460 women aged 50 to 69 screened by the Ontario Breast Screening Program, offering eligible women screening with mammography and clinical breast examination, were examined. Women were followed for up to 12 months after their screening examination. Family history definitions were based on the number of affected first degree relatives and their ages at diagnosis. Multivariate analysis was conducted to adjust for potential confounding variables. RESULTS: Interval cancer rates increased across family history groups and were greatest in women with a strong family history. The rate ratio (RR) for interval cancer rate in women with a strong family history compared to women without a family history approached significance (RR=2.28, 95% confidence interval (CI) 0.97-5.34), while for women with a moderate family history it did not (RR=1.37, 95% CI 0.62-3.04). A slightly but not significantly lower sensitivity was observed in women with a strong family history compared to women without a family history. There was little variation in specificity across family history groups. CONCLUSIONS: Screening was able to detect a large proportion of invasive breast cancers in women with a family history, indicating their potential to benefit from regular breast cancer screening. However, due to increased interval cancer rates, screening with one year intervals may be important even in an older population of women with a family history. PMID- 15830145 TI - Effect of organ-specific fibroblasts on proliferation and differentiation of breast cancer cells. AB - Breast carcinomas contain both tumor cells and stromal cells, including fibroblasts, endothelial cells, and lymphocytes. Proliferation of breast cancer cells may be controlled partly by mesenchymal cells, especially fibroblasts. We studied effects of fibroblasts on tumorigenicity and histologic features of breast cancer cells vivo, and analyzed fibroblast-produced growth-promoting factors in vitro. Breast carcinoma cells from four lines, and fibroblasts from lines obtained from skin and breast tissue of four patients with breast cancer were used. A suitable number of breast tumor cells and fibroblasts were inoculated subcutaneously into nude mice; resulting tumors were examined. Then conditioned medium from fibroblasts was added to cultures of breast cancer cells to study growth effects, and growth-promoting factors from breast fibroblasts were analyzed. Co-inoculation of breast cancer cells with breast fibroblasts into mice significantly increased tumorigenicity and tumor size beyond those obtained with breast cancer cells alone. Histologically, tumors resulting from co inoculation with breast fibroblasts showed a scirrhous pattern with extensive fibrosis, while those formed by breast cancer cells injected alone or co inoculation with skin fibroblasts showed a solid pattern. Medium from breast fibroblasts significantly increased breast cancer cell growth in vitro, while the various skin fibroblasts did not all show this effect. Structural and functional interactions between organ-specific fibroblasts and breast cancer cells may importantly regulate breast cancer growth and progression. PMID- 15830146 TI - Should oral bisphosphonates be standard of care in women with early breast cancer? AB - There is increasing evidence that oral clodronate has important clinical benefits in early breast cancer. Longer-term follow-up data from two out of three randomized studies have shown that adjuvant clodronate improves survival. This article outlines the results of these studies and discusses the potential role for other bisphosphonates, such as ibandronate and zoledronic acid, in the adjuvant treatment of breast cancer. PMID- 15830147 TI - Lumpectomy and tamoxifen alone without additional radiotherapy for women 70 years of age or older with estrogen receptor-positive breast cancer. PMID- 15830148 TI - DNA polymerase gene locus of Cercopithecine herpesvirus 1 is a suitable target for specific and rapid identification of viral infection by PCR technology. AB - The family Herpesviridae comprises at least 100 herpesviruses. Numerous human and animal pathogenic herpesviruses have been identified so far, including Cercopithecine herpesvirus 1 (CeHV-1). This virus is a member of the subfamily Alphaherpesvirinae and is the most hazardous herpesvirus to man. CeHV-1 is also known as B-virus or monkey B virus and as Herpesvirus simiae. In order to gain more genetic information, the viral DNA polymerase (DPOL) gene was identified using polymerase chain reaction (PCR) and DNA nucleotide sequence analysis. The deduced amino acid sequence contains the motifs and signatures that are typical for the B-family of DPOLs. The DPOL gene of CeHV-1 was found to be a suitable target for the specific and rapid identification of the Cercopithecine herpesvirus 1 infection by PCR technology. Comparative analysis of the DNA sequences of the DPOL gene loci of CeHV-1, Human herpesvirus 1 and 2 (HHV-1 and HHV-2), and other herpesviruses was carried out for determination of unique genomic regions of the individual DPOL genes. A primer set of 12 primers was used for screening the DNA of CeHV-1, HHV-1, and HHV-2 by detailed PCR. It was found that six out of twelve primer combinations are able to detect specifically the CeHV-1 genome without cross reactivity with the genome of HHV-1 and/or HHV-2. The specificity of the individual amplified DNA fragments was confirmed by DNA nucleotide sequence analysis. The results of these studies indicate that the six primer combinations of the specific CeHV-1 DPOL primer set is the method of choice for a rapid, precise and specific identification of a CeHV-1 infection by PCR. Due to the fact that this specific CeHV-1 DPOL primer set does not amplify any DNAs of HHV-1 or HHV-2 genome this technology is stressing and can be successfully used unlimited and more credible in all laboratories with PCR technical facility routinely for detection of a CeHV-1 infection in vivo or in vitro. PMID- 15830149 TI - EBV infection induces expression of the transcription factors ATF-2/c-Jun in B lymphocytes but not in B-CLL cells. AB - B cell type chronic lymphocytic leukaemia (B-CLL) cells carry the Epstein-Barr virus (EBV) receptor CD21 and can be infected in vitro with the virus. The infected cells exhibit an unusual EBV program, they express the nuclear proteins but not latent membrane protein 1 (LMP-1). Similar cells were encountered in lymphoid tissues of infectious mononucleosis (IM) patients and in lymphoproliferations of immunosuppressed patients. EBV infected B-CLL cells can be regarded as model for this viral program. In B cells the regulation of LMP-1 is executed mainly by EBV encoded nuclear antigen 2 (EBNA-2), interacting with several cellular proteins and these complexes bind to specific sequences in the LMP-1 promoter. ATF2 and c-Jun were shown to be among the interacting partners of EBNA-2. These molecules can be detected in experimentally infected B lymphocytes. We found c-Jun and/or phosphorylated ATF-2 (p-ATF-2) expression in some B-CLL ex vivo samples. They disappeared or their expression declined promptly in explanted cells, even if they were infected with EBV in vitro. Activation of the infected B CLL cells by exposure to CD40L was accompanied by p-ATF-2 and c-Jun but not by LMP-1 expression. In one of three clones tested, subsequent treatment with histone deacetylase inhibitors (HDACi), TSA or n-butyrate, could induce LMP-1. Treatment with phorbol-12, 13-dibutyrate (PDB) induced LMP-1 expression in three of four clones. Neither the HDACi nor the PDB treated cells survived. PMID- 15830150 TI - Analysis of the complete genome sequence of avian metapneumovirus subgroup C indicates that it possesses the longest genome among metapneumoviruses. PMID- 15830151 TI - Complete nucleotide sequence of the RNA-2 of grapevine deformation and Grapevine Anatolian ringspot viruses. AB - The nucleotide sequence of RNA-2 of Grapevine Anatolian ringspot virus (GARSV) and Grapevine deformation virus (GDefV), two recently described nepoviruses, has been determined. These RNAs are 3753 nt (GDefV) and 4607 nt (GARSV) in size and contain a single open reading frame encoding a polyprotein of 122 kDa (GDefV) and 150 kDa (GARSV). Full-length nucleotide sequence comparison disclosed 71-73% homology between GDefV RNA-2 and that of Grapevine fanleaf virus (GFLV) and Arabis mosaic virus (ArMV), and 62-64% homology between GARSV RNA-2 and that of Grapevine chrome mosaic virus (GCMV) and Tomato black ring virus (TBRV). As previously observed in other nepoviruses, the 5' non-coding regions of both RNAs are capable of forming stem-loop structures. Phylogenetic analysis of the three proteins encoded by RNA-2 (i.e. protein 2A, movement protein and coat protein) confirmed that GDefV and GARSV are distinct viruses which can be assigned as definitive species in subgroup A and subgroup B of the genus Nepovirus, respectively. PMID- 15830152 TI - Genetic characterization of rabies virus isolates in Korea. AB - In investigation of the genetic characteristics of rabies viruses in Korea, the nucleotide and deduced amino acid sequences of the nucleoprotein (N) gene were determined in four Korean rabies virus strains obtained from dogs and raccoons, and were compared with published sequences for non-Korean rabies viruses. Three Korean rabies virus isolates had identical nucleotide sequences, and the fourth differed at only one nucleotide position. The Korean virus isolates had 84.5 92.0% nucleotide sequence similarity and 94.0-99.2% amino acid sequence similarity with non-Korean rabies virus isolates. In a phylogenetic tree based on partial nucleotide sequences of the N gene, the Korean rabies viruses formed a single cluster closely related to Arctic rabies viruses (FXCAN, 9141RUS, and 94260NEP). However, they were divergent from other Asian rabies viruses (94256SRL, 8677MAL, ChiNo.7, Phil 12301, and Mdn1278). PMID- 15830153 TI - Characterization of p24 gene of Spodoptera litura multicapsid nucleopolyhedrovirus. AB - Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) p24 gene is 753 bp long, potentially encoding 244 amino acids with a predicted molecular weight of 27.3 kDa. Homology analysis indicated that SpltMNPV P24 has 20-36% amino acid identity with that of other known baculoviruses. RT-PCR results showed that the p24 gene is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter sequence (ATAAG). Western blot analysis of extracts from SpltMNPV-infected S. litura cells detected a specific 28 kDa protein, and this protein was not N-glycosylated. Structural localization revealed that SpltMNPV P24 was associated with the nucleocapsid of occlusion-derived virus (ODV) as a complex form of 83 kDa. PMID- 15830154 TI - Reliable classification and recombination analysis of porcine endogenous retroviruses. AB - Prevention of cross-species infection with porcine endogenous retroviruses (PERV) is crucial for xenotransplantation. Previous studies described the potential risk of infection for the PERV gamma1 subfamilies A, B and C. Replication competent PERV gamma1 proviruses designated to a particular subfamily and hybrid viruses originating from retroviral recombination events between the subfamilies were observed. Future pig genome sequencing projects will reveal multiple novel PERV proviruses from additional breeds and animals. Evaluation of these viral genomes has to be carried out to assess the potential risk of retroviral cross-species infection. In this study, we tested common sequence comparison methods for the classification of PERV sequences and the detection of hybrid clones. The examination of the polymorphic nucleotide positions was found to be the most suitable procedure. We describe a fast and simple method using bioinformatic software tools which can also be applied to analogous analyses of other viral genomes. PMID- 15830155 TI - Quasispecies nature of an unusual avian paramyxovirus type-1 isolated from pigeons. AB - An avian paramyxovirus type-1 (APMV-1) was classified as virulent according to its Intra Cerebral Pathogenicity Index (ICPI), but as avirulent according to the motif of its F protein cleavage site. Although this atypical APMV-1 was isolated from sick, unvaccinated pigeons, it was not grouped with pigeon variants regarding its antigenic and genetic characterisation. We analysed its quasispecies nature by cloning and sequencing parts of the genome in three different genes to evaluate if heterogeneity might explain the difference observed between the ICPI and the F protein cleavage site motif. Two distinct sub populations were detected in the phosphoprotein gene. In the fusion protein gene, two clones were found to be related to typical pigeon variants in the hypervariable domain. PMID- 15830156 TI - Phylogenetic analysis of the DNA polymerase gene of a novel alphaherpesvirus isolated from an Indian Gyps vulture. AB - The DNA polymerase gene of a novel herpesvirus, vulture herpesvirus (VHV), isolated from an Indian Gyps vulture was completely sequenced using primer walking and transposon insertion strategies. DNA sequencing analysis revealed a single open reading frame (ORF) of 3660 nucleotides (53% G-C content) able to encode 1219 amino acids. Identification was based on a nucleotide sequence identity of approximately 50% to other herpesvirus sequences found in Genbank. Nine motifs were identified that are conserved amongst all known herpesviruses and are found within the 3'-5' exonuclease and DNA binding functional domains of the DNA polymerase enzyme. Phylogenetic analysis using Clustal W with neighbour joining revealed VHV to group within the subfamily Alphaherpesvirinae, more closely related to the avian herpesviruses than to those of other species. Partial sequence data also revealed VHV to contain other genes fundamental to the structure and replication of all herpesvirus genomes. A Real Time PCR Taqman assay specific for the VHV DNA polymerase gene was designed to detect the presence of VHV genomic material in post mortem tissue samples from diseased birds. Positive tissues included the spleen, rectum, thymus, kidney and brain. A herpesvirus specific to vultures may pose a threat to the management of captive breeding programs being established to assist the survival of wild populations of Gyps vultures. PMID- 15830157 TI - Molecular phylogenetic analysis of Iranian HDV complete genome. AB - Hepatitis D (delta) virus (HDV) is a subviral pathogen agent and a satellite of Hepatitis B virus. Three distinct genotypes are described for HDV; genotype I is distributed worldwide but other genotypes appear to be more restricted geographically. In the present study, the entire nucleotide sequence of an HDV isolate from an Iranian patient (IR-1) was obtained using twelve pairs of primers to amplify six overlapping fragments covering the whole HDV genome by RT-nested PCR. Phylogenetic and pairwise alignments were done on this new isolate to determine IR-1 position among other isolates. Our results indicate that IR-1 contains 1676 nucleotides encoding 214 a.a. of the hepatitis delta antigen (HDAg). This new isolate belongs to genotype I with most sequence similarity to an Italian HDV isolate (92.6%). At amino acid level, predicted HDAg sequence of IR-1 revealed the most homology with those of Italian and Lebanese isolates. Data analysis confirmed genetic variability and heterogeneity of the HDV species isolated from different geographical areas. PMID- 15830158 TI - Molecular cloning and phylogenetic analysis of ORF7 region of chinese isolate TH 98 from transmissible gastroenteritis virus. AB - Genomic RNA was extracted from a Chinese isolate of porcine transmissible gastroenteritis virus (TGEV) designated TH-98. Employing RT-PCR technique to amplify ORF7 sequence of TGEV, which located at the 3' end of TGEV genome and is poorly understood functionally so far. A recombinant named pPROEX HTc-hp was constructed via inserting ORF7 gene into prokaryotic expression vector pPROEX HTc. The recombinant was sequenced and compared the DNA and its deduced amino acid (aa) sequences with that of some reference strains after restriction endonuclease and PCR analysis. The ORF7 gene named hp gene (Genbank accession number: AY337931) consists of 237 bp in length encoding a hydrophobic protein (HP) of 78 aa with a molecular weight of 9.1 kDa. The sequences of hp gene and Hp protein share 89%-97% and 87%-96% homologous identities compared with 11 TGEV reference strains derived from other regions or countries respectively, which revealed that there are significant variation within-strains, even though the ORF7 region is relatively conservative. In addition, a phylogenetic tree based on these ORF7 DNA sequences was generated, and the tree topology suggests that possible recombination events happened in the evolutionary history of TGEV. PMID- 15830159 TI - Physical and partial genetic map of Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV) genome. AB - A Nicaraguan isolate of Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV) is undergoing field trials for control of this pest in the Americas. This isolate is composed of multiple genotypes, some of which are deletion mutants. Identification of the genetic changes in deleted genotypes cannot be accomplished without the construction of a detailed physical map. In the present study, combinations of restriction endonuclease analysis and Southern blot analysis was performed. This map was refined by sequencing the termini of cloned restriction fragments. The SfMNPV genome was estimated to be 129.3 kb, 8 kb larger than the previously characterized Sf-2 variant from the United States, due to a deletion between 14.8 and 21.0 m.u. in the physical map described in this study. A total of 27.92 kb were sequenced, which represented 21.5% of the whole genome and included 38 ORFs. Comparison with other sequenced baculoviruses revealed that SfMNPV displayed the highest sequence identity (66%) and gene arrangement (78%) with Spodoptera exigua MNPV, sharing 36 putative ORFs. In addition, the genome organization was similar to that of SeMNPV, with minor differences. Phylogenetic analysis confirmed the close relatedness between SeMNPV and SfMNPV, suggesting they evolved from a common ancestor. PMID- 15830162 TI - Coexistence of tonic spiking oscillations in a leech neuron model. AB - The leech neuron model studied here has a remarkable dynamical plasticity. It exhibits a wide range of activities including various types of tonic spiking and bursting. In this study we apply methods of the qualitative theory of dynamical systems and the bifurcation theory to analyze the dynamics of the leech neuron model with emphasis on tonic spiking regimes. We show that the model can demonstrate bi-stability, such that two modes of tonic spiking coexist. Under a certain parameter regime, both tonic spiking modes are represented by the periodic attractors. As a bifurcation parameter is varied, one of the attractors becomes chaotic through a cascade of period-doubling bifurcations, while the other remains periodic. Thus, the system can demonstrate co-existence of a periodic tonic spiking with either periodic or chaotic tonic spiking. Pontryagin's averaging technique is used to locate the periodic orbits in the phase space. PMID- 15830163 TI - The onset and extinction of neural spiking: a numerical bifurcation approach. AB - We study the onset of neural spiking when the equilibrium rest state loses stability by the change of a critical parameter, the applied current. In the case of the well-known Morris-Lecar model, we start from a complete numerical study of the bifurcation diagram in the most relevant two-parameter range. This diagram includes all equilibrium and limit cycle bifurcations, thus correcting and completing earlier studies. We discuss and classify the behavior of the spiking orbits, when increasing or decreasing the applied current. A complete classification can be extracted from the complete bifurcation diagram. It is based on three components: bifurcation type of the equilibrium at the loss of stability, subcritical behavior in the limit of decreasing the applied current and supercritical behavior in the limit of increasing the applied current. PMID- 15830164 TI - The possible role of spike patterns in cortical information processing. AB - When the same visual stimulus is presented across many trials, neurons in the visual cortex receive stimulus-related synaptic inputs that are reproducible across trials (S) and inputs that are not (N). The variability of spike trains recorded in the visual cortex and their apparent lack of spike-to-spike correlations beyond that implied by firing rate fluctuations, has been taken as evidence for a low S/N ratio. A recent re-analysis of in vivo cortical data revealed evidence for spike-to-spike correlations in the form of spike patterns. We examine neural dynamics at a higher S/N in order to determine what possible role spike patterns could play in cortical information processing. In vivo-like spike patterns were obtained in model simulations. Superpositions of multiple sinusoidal driving currents were especially effective in producing stable long lasting patterns. By applying current pulses that were either short and strong or long and weak, neurons could be made to switch from one pattern to another. Cortical neurons with similar stimulus preferences are located near each other, have similar biophysical properties and receive a large number of common synaptic inputs. Hence, recordings of a single neuron across multiple trials are usually interpreted as the response of an ensemble of these neurons during one trial. In the presence of distinct spike patterns across trials there is ambiguity in what would be the corresponding ensemble, it could consist of the same spike pattern for each neuron or a set of patterns across neurons. We found that the spiking response of a neuron receiving these ensemble inputs was determined by the spike pattern composition, which, in turn, could be modulated dynamically as a means for cortical information processing. PMID- 15830165 TI - Beyond two-cell networks: experimental measurement of neuronal responses to multiple synaptic inputs. AB - Oscillations of large populations of neurons are thought to be important in the normal functioning of the brain. We have used phase response curve (PRC) methods to characterize the dynamics of single neurons and predict population dynamics. Our past experimental work was limited to special circumstances (e.g., 2-cell networks of periodically firing neurons). Here, we explore the feasibility of extending our methods to predict the synchronization properties of stellate cells (SCs) in the rat entorhinal cortex under broader conditions. In particular, we test the hypothesis that PRCs in SCs scale linearly with changes in synaptic amplitude, and measure how well responses to Poisson process-driven inputs can be predicted in terms of PRCs. Although we see nonlinear responses to excitatory and inhibitory inputs, we find that models based on weak coupling account for scaling and Poisson process-driven inputs reasonably accurately. PMID- 15830166 TI - Action potential onset dynamics and the response speed of neuronal populations. AB - The result of computational operations performed at the single cell level are coded into sequences of action potentials (APs). In the cerebral cortex, due to its columnar organization, large number of neurons are involved in any individual processing task. It is therefore important to understand how the properties of coding at the level of neuronal populations are determined by the dynamics of single neuron AP generation. Here, we analyze how the AP generating mechanism determines the speed with which an ensemble of neurons can represent transient stochastic input signals. We analyze a generalization of the theta-neuron, the normal form of the dynamics of Type-I excitable membranes. Using a novel sparse matrix representation of the Fokker-Planck equation, which describes the ensemble dynamics, we calculate the transmission functions for small modulations of the mean current and noise noise amplitude. In the high-frequency limit the transmission function decays as omega(-gamma), where gamma surprisingly depends on the phase theta(s) at which APs are emitted. If at theta(s) the dynamics is insensitive to external inputs, the transmission function decays as (i) omega(-3) for the case of a modulation of a white noise input and as (ii) omega(-2) for a modulation of the mean input current in the presence of a correlated and uncorrelated noise as well as (iii) in the case of a modulated amplitude of a correlated noise input. If the insensitivity condition is lifted, the transmission function always decays as omega(-1), as in conductance based neuron models. In a physiologically plausible regime up to 1 kHz the typical response speed is, however, independent of the high-frequency limit and is set by the rapidness of the AP onset, as revealed by the full transmission function. In this regime modulations of the noise amplitude can be transmitted faithfully up to much higher frequencies than modulations in the mean input current. We finally show that the linear response approach used is valid for a large regime of stimulus amplitudes. PMID- 15830167 TI - Dynamics of the instantaneous firing rate in response to changes in input statistics. AB - We review and extend recent results on the instantaneous firing rate dynamics of simplified models of spiking neurons in response to noisy current inputs. It has been shown recently that the response of the instantaneous firing rate to small amplitude oscillations in the mean inputs depends in the large frequency limit f on the spike initiation dynamics. A particular simplified model, the exponential integrate-and-fire (EIF) model, has a response that decays as 1/f in the large frequency limit and describes very well the response of conductance-based models with a Hodgkin-Huxley type fast sodium current. Here, we show that the response of the EIF instantaneous firing rate also decays as 1/f in the case of an oscillation in the variance of the inputs for both white and colored noise. We then compute the initial transient response of the firing rate of the EIF model to a step change in its mean inputs and/or in the variance of its inputs. We show that in both cases the response speed is proportional to the neuron stationary firing rate and inversely proportional to a 'spike slope factor' Delta(T) that controls the sharpness of spike initiation: as 1/Delta(T) for a step change in mean inputs, and as 1/Delta(T) (2) for a step change in the variance in the inputs. PMID- 15830168 TI - Short-term synaptic plasticity orchestrates the response of pyramidal cells and interneurons to population bursts. AB - The synaptic drive from neuronal populations varies considerably over short time scales. Such changes in the pre-synaptic rate trigger many temporal processes absent under steady-state conditions. This paper examines the differential impact of pyramidal cell population bursts on post-synaptic pyramidal cells receiving depressing synapses, and on a class of interneuron that receives facilitating synapses. In experiment a significant shift of the order of one hundred milliseconds is seen between the response of these two cell classes to the same population burst. It is demonstrated here that such a temporal differentiation of the response can be explained by the synaptic and membrane properties without recourse to elaborate cortical wiring schemes. Experimental data is first used to construct models of the two types of dynamic synaptic response. A population based approach is then followed to examine analytically the temporal synaptic filtering effects of the population burst for the two post-synaptic targets. The peak-to-peak delays seen in experiment can be captured by the model for experimentally realistic parameter ranges. It is further shown that the temporal separation of the response is communicated in the outgoing action potentials of the two post-synaptic cells: pyramidal cells fire at the beginning of the burst and the class of interneuron receiving facilitating synapses fires at the end of the burst. The functional role of such delays in the temporal organisation of activity in the cortical microcircuit is discussed. PMID- 15830169 TI - Surprising effects of synaptic excitation. AB - Typically, excitatory synaptic coupling is thought of as an influence that accelerates and propagates firing in neuronal networks. This paper reviews recent results explaining how, contrary to these expectations, the presence of excitatory synaptic coupling can drastically slow oscillations in a network and how localized, sustained activity can arise in a network with purely excitatory coupling, without sustained inputs. These two effects stem from interactions of excitatory coupling with two different forms of intrinsic neuronal dynamics, and both serve to highlight the fact that the influence of synaptic coupling in a network depends strongly on the intrinsic properties of cells in the network. PMID- 15830170 TI - Two-cell to N-cell heterogeneous, inhibitory networks: precise linking of multistable and coherent properties. AB - Inhibitory networks are now recognized as being the controllers of several brain rhythms. However, experimental work with inhibitory cells is technically difficult not only because of their smaller percentage of the neuronal population, but also because of their diverse properties. As such, inhibitory network models with tight links to the experimental data are needed to understand their contributions to population rhythms. However, mathematical analyses of network models with more than two cells is challenging when the cellular models involve biophysical details. We use bifurcation analyses and simulations to show that two-cell analyses can quantitatively predict N-cell (N = 20, 50, 100) network dynamics for heterogeneous, inhibitory networks. Interestingly, multistable states in the two-cell system are manifest as different and distinct coherent network patterns in the N-cell networks for the same parameter sets. PMID- 15830171 TI - [Use of phytopharmaceutical agents in dermatology. Indications, therapeutic approaches and side effects]. AB - In the recent years there has been increasing interest in herbal therapy including its use for treatment of dermatologic disorders. In Germany, a regulatory authority known as "Commission E" performed an extensive review of common phytotherapeutic (1987-1995) agents indicating quality of evidence, clinical efficacy and therapeutic usefulness producing a "positive monograph" indicating agents supported by evidence and likely to be useful. This article explores the therapeutic use of several of these and other currently popular plants suitable for dermatologic indications; therapeutic hints including phytotherapeutic products and formulations are given and side effects, in particular toxic and allergic skin reactions, are discussed. PMID- 15830172 TI - [Postmenopausal and steroid-induced osteoporosis guideline-orientated prevention and treatment]. AB - Prevention of osteoporosis and osteoporotic fractures consists of non-drug and drug therapy. Components of non-drug therapy include improvement of muscle strength and coordination, treatment of modifiable causes of falls, a diet rich in calcium and sufficient in calories, adequate supply of vitamin D and a careful approach towards drugs known to increase falls or osteoporosis. Assessment of clinical risk factors in combination with bone mineral density measurements can identify persons at high risk of fracture who benefit most from pharmacological treatment. Particular attention should be paid to patients with previous fragility fractures and long-term oral glucocorticoid therapy. PMID- 15830173 TI - [Possible ways of managing cardiovascular prevention: polypharmacy, additional payment or application of evidence based medicine?]. AB - The financial balance of the health care system has changed dramatically due to a longer life expectancy and improved treatment options in elderly patients. More than 80% of cardiovascular events are lifestyle related and potentially preventable. Lifestyle modification is therefore the causal approach to decrease cardiovascular events. Improvement of nutrition and activity habits and prevention of cigarette smoking should start in the kindergarten, school and later at the workplace. A co-operation between medical societies and government institutions is necessary to achieve a population wide modification of lifestyle habits to lower the incidence of cardiovascular events in the population. Individual risk stratification is the basis for pharmacological prevention of cardiovascular events. The concept of the polypill has to be tested in controlled randomised studies. PMID- 15830174 TI - [Osteoporosis in traumatology. Prevalence and management]. AB - BACKGROUND: A fracture occurring in osteoporosis is a sentinel event but very rarely leads to bone mineral density (BMD) measurement or sufficient drug therapy. We designed an algorithm to evaluate BMD in older fracture patients and tested it for sustainability as well as acceptance among trauma/orthopedic surgeons. METHODS: For a 1-year period a prospective BMD test was carried out in women older than 50 and men older than 75 years of age with fractures. The commencement and conduction of therapy during the initial hospital stay and rehabilitation were also analyzed. RESULTS: From 228 members of the eligible age groups, 169 patients (74.1%) underwent BMD measurement. According to the WHO definition 71.6% showed reduced BMD in terms of osteoporosis and 24.3% in terms of osteopenia. In 84% therapy was started during initial hospital stay in 74.4% conducted during rehabilitation. CONCLUSIONS: The vast majority of older patients exhibited reduced BMD as a substantial underlying cause of their fracture. A standardized clinical plan can help to identify and treat most patients with fragility fractures. PMID- 15830175 TI - [Out-of-body experience as possible seizure symptom in a patient with a right parietal lesion]. AB - Out-of-body experiences are reported in a variety of diseases and physiologic conditions. We report a 44-year-old patient with epigastric auras, psychomotor and grand mal seizures, and paroxysmal experiences during which he believed to have left his body and seen himself from the outside. Electroencephalography showed a right temporal and right parietal seizure pattern on several occasions, and a lesion in the right parietal lobe was detected by cranial magnetic resonance imaging. Histological examination showed a pleomorphic xanthoastrocytoma. Following extirpation of the lesion, the patient has been seizure-free for 5 years. PMID- 15830176 TI - Comment to: Jeffcoate W J, van Houtum W H (2004) Amputation as a marker of the quality of foot care in diabetes. Diabetologia 47:2051-2058. PMID- 15830177 TI - Molecular genetics and phenotypic characteristics of MODY caused by hepatocyte nuclear factor 4alpha mutations in a large European collection. AB - AIMS/HYPOTHESIS: Heterozygous mutations in the gene of the transcription factor hepatocyte nuclear factor 4alpha (HNF-4alpha) are considered a rare cause of MODY with only 14 mutations reported to date. The description of the phenotype is limited to single families. We investigated the genetics and phenotype of HNF 4alpha mutations in a large European Caucasian collection. METHODS: HNF-4alpha was sequenced in 48 MODY probands, selected for a phenotype of HNF-1alpha MODY but negative for HNF-1alpha mutations. Clinical characteristics and biochemistry were compared between 54 HNF-4alpha mutation carriers and 32 familial controls from ten newly detected or previously described families. RESULTS: Mutations in HNF-4alpha were found in 14/48 (29%) probands negative for HNF-1alpha mutations. The mutations found included seven novel mutations: S34X, D206Y, E276D, L332P, I314F, L332insCTG and IVS5nt+1G>A. I314F is the first reported de novo HNF-4alpha mutation. The average age of diagnosis was 22.9 years with frequent clinical evidence of sensitivity to sulphonylureas. Beta cell function, but not insulin sensitivity, was reduced in diabetic mutation carriers compared to control subjects (homeostasis model assessment of beta cell function 29% p<0.001 vs controls). HNF-4alpha mutations were associated with lower apolipoprotein A2 (p=0.001), A1 (p=0.04) and total HDL-cholesterol (p=0.02) than in control subjects. However, in contrast to some previous reports, levels of triglycerides and apolipoprotein C3 were normal. CONCLUSIONS/INTERPRETATION: HNF-4alpha mutations are common when no HNF-1alpha mutation is found in strictly defined MODY families. The HNF-4alpha clinical phenotype and beta cell dysfunction are similar to HNF-1alpha MODY and are associated with reduced apolipoprotein A2 levels. We suggest that sequencing of HNF-4alpha should be performed in patients with clinical characteristics of HNF-1alpha MODY in whom mutations in HNF-1alpha are not found. PMID- 15830178 TI - Clinical significance of the physicochemical properties of LDL in type 2 diabetes. AB - Atherosclerosis is the leading cause of death in type 2 diabetes. LDL cholesterol and atherosclerosis are related, both in healthy people and those with diabetes; however, people with diabetes are more prone to atheroma, even though their LDL cholesterol levels are similar to those in their non-diabetic peers. This is because LDL particles are modified in the presence of diabetes to become more atherogenic. These modifications include glycation in response to high plasma glucose levels; oxidative reactions mediated by increased oxidative stress; and transfer of cholesterol ester, which makes the particles smaller and denser. The latter modification is strongly associated with hypertriglyceridaemia. Oxidatively and non-oxidatively modified LDL is involved in plaque formation, and may thus contribute to the accelerated atherosclerosis. This review discusses the techniques currently used to determine the physicochemical properties of LDL, and examines the evidence that modification of these properties plays a role in the accelerated atherosclerosis associated with type 2 diabetes. PMID- 15830179 TI - Hypoadiponectinaemia and high risk of type 2 diabetes are associated with adiponectin-encoding (ACDC) gene promoter variants in morbid obesity: evidence for a role of ACDC in diabesity. AB - AIMS/HYPOTHESIS: Morbid obesity (BMI>40 kg/m(2)) affecting 0.5-5% of the adult population worldwide is a major risk factor for type 2 diabetes. We aimed to elucidate the genetic bases of diabetes associated with obesity (diabesity), and to analyse the impact of corpulence on the effects of diabetes susceptibility genes. METHODS: We genotyped known single nucleotide polymorphisms (SNPs) in the adiponectin-encoding adipocyte C1q and collagen-domain-containing (ACDC) gene ( 11,391G>A, -11,377C>G, +45T>G and +276G>T), the peroxisome proliferator-activated receptor gamma (PPARG) Pro12Ala SNP and ACDC exon 3 variants in 703 French morbidly obese subjects (BMI 47.6+/-7.4 kg/m(2)), 808 non-obese subjects (BMI<30 kg/m(2)) and 493 obese subjects (30< or =BMI<40 kg/m(2)). RESULTS: Two 5'-ACDC SNPs -11,391G>A, -11,377C>G were associated with adiponectin levels (p=0.0003, p=0.008) and defined a "low-level" haplotype associated with decreased adiponectin levels (p=0.0002) and insulin sensitivity (p=0.01) and with a risk of type 2 diabetes that was twice as high (p=0.002). In contrast, the prevalence of the PPARG Pro12Ala was identical in diabetic and normoglycaemic morbidly obese subjects. The PPARG Pro12 allele only displayed a trend of association with type 2 diabetes in the non-obese group. ACDC exon 3 variants were associated with type 2 diabetes in the non-obese group only (odds ratio 7.85, p<0.0001). In contrast, the 5'-ACDC "low-level" haplotype was associated with type 2 diabetes in obese and morbidly obese subjects (odds ratio 1.73 and 1.92) but not in non-obese individuals. CONCLUSIONS/INTERPRETATION: These data clarify the contribution of the 5'-ACDC SNPs to the risk of diabesity. Their interaction with corpulence suggests for the first time a different genetic profile of type 2 diabetes in morbidly obese patients compared with in less obese individuals. PMID- 15830180 TI - Effective treatment of symptomatic diabetic polyneuropathy by high-frequency external muscle stimulation. AB - AIMS/HYPOTHESIS: Diabetic distal symmetrical sensory polyneuropathy (DSP) affects 20-30% of diabetic patients. Transcutaneous electrical nerve stimulation (TENS) and electrical spinal cord stimulation have been proposed as physical therapies. We performed a controlled, randomised pilot trial to compare the effects of high frequency external muscle stimulation (HF) with those of TENS in patients with symptomatic DSP. METHODS: Patients with type 2 diabetes and DSP (n=41) were randomised to receive treatment with TENS or HF using strata for non-painful (n=20) and painful sensory symptoms (n=21). Both lower extremities were treated for 30 min daily for three consecutive days. The patients' degree of symptoms and pain were graded daily on a scale of one to ten, before, during and 2 days after treatment termination. Responders were defined by the alleviation of one or more symptoms by at least three points. RESULTS: The two treatment groups were similar in terms of baseline characteristics, such as age, duration of diabetes, neurological symptoms scores and neurological disability scores. The responder rate was significantly higher (p<0.05) in the HF group (80%, 16 out of 20) than in the TENS group (33%, seven out of 21). Subgroup analysis revealed that HF was more effective than TENS in relieving the symptoms of non-painful neuropathy (HF: 100%, seven out of seven; TENS: 44%, four out of nine; p<0.05) and painful neuropathy (HF: 69%, nine out of 13; TENS: 25%, three out of 12; p<0.05). The responders did not differ in terms of the reduction in mean symptom intensity during the trial. CONCLUSIONS/INTERPRETATION: This pilot study shows, for the first time, that HF can ameliorate the discomfort and pain associated with DSP, and suggests that HF is more effective than TENS. External muscle stimulation offers a new therapeutic option for DSP. PMID- 15830181 TI - Heterogeneity in limb fatty acid kinetics in type 2 diabetes. AB - AIMS/HYPOTHESIS: In order to test the hypothesis that disturbances in skeletal muscle fatty acid metabolism with type 2 diabetes are not equally present in the upper and lower limbs, we studied fatty acid kinetics simultaneously across the arm and leg of type 2 diabetic patients (n=6) and matched control subjects (n=7) for 5 h under baseline conditions and during a 4-h hyperinsulinaemic-euglycaemic clamp. METHODS: Limb fatty acid kinetics was determined by means of continuous [U (13)C]palmitate infusion and measurement of arteriovenous differences. RESULTS: The systemic palmitate rate of appearance was 3.6+/-0.4 and 2.7+/-0.3 micromol.kg lean body mass(-1).min(-1) and decreased during the clamp by 26% (p=0.04) and 43% (p<0.01) in the diabetic patients and in the control subjects respectively. At baseline, palmitate uptake across the arm was similar in the two groups, whereas leg palmitate uptake was lower than in the arm in the diabetic patients. During the clamp, palmitate uptake decreased in the arm (-48%, p=0.02) and the leg ( 38%, p=0.04) of the control subjects, whereas it decreased in the arm (-30%, p=0.04) but not in the leg of the diabetic patients. Similarly, during the clamp palmitate release was substantially suppressed in the arm (-47%, p<0.01) and the leg of the control subjects (-45%, p<0.01), but only in the arm of the diabetic patients (-45%, p<0.01). CONCLUSIONS/INTERPRETATION: The present data indicate that type 2 diabetes is characterised by heterogeneity in the dysregulation of skeletal muscle fatty acid metabolism, with only the leg, but not the arm, showing an impairment of fatty acid kinetics at baseline and during a hyperinsulinaemic-euglycaemic clamp causing a physiological increase in insulin concentration. PMID- 15830182 TI - Angiotensin-I converting enzyme insertion/deletion polymorphism and its association with diabetic nephropathy: a meta-analysis of studies reported between 1994 and 2004 and comprising 14,727 subjects. AB - AIMS/HYPOTHESIS: The ACE insertion/deletion polymorphism has been examined for association with diabetic nephropathy over the past decade with conflicting results. To clarify this situation, we conducted a comprehensive meta-analysis encompassing all relevant studies that were published between 1994 and 2004 and investigated this potential genetic association. METHODS: A total of 14,727 subjects from 47 studies was included in this meta-analysis. Cases (n=8,663) were type 1 or 2 diabetic subjects with incipient (microalbuminuria) or advanced diabetic nephropathy (proteinuria, chronic renal failure, end-stage renal disease). Control subjects (n=6,064) were predominantly normoalbuminuric. RESULTS: No obvious publication bias was detected. Using a minimal-case definition based on incipient diabetic nephropathy, subjects with the II genotype had a 22% lower risk of diabetic nephropathy than carriers of the D allele (pooled odds ratio [OR]=0.78, 95% CI=0.69-0.88). While there was a reduced risk of diabetic nephropathy associated with the II genotype among Caucasians with either type 1 or type 2 diabetes, the association was most marked among type 2 diabetic Asians (Chinese, Japanese, Koreans) (OR=0.65, 95% CI=0. 51-0.83). This OR is significantly different from the OR of 0.90 (95% CI= 0.78-1.04) that was obtained for type 2 diabetic Caucasians (p=0.019). Using a stricter case definition based on advanced diabetic nephropathy, a comparable risk reduction of 24-32% was observed among the three subgroups, although statistical significance was reached only among Asians. CONCLUSIONS/INTERPRETATION: The results of our meta-analysis support a genetic association of the ACE Ins/Del polymorphism with diabetic nephropathy. These findings may have implications for the management of diabetic nephropathy using ACE inhibitors especially among type 2 diabetic Asians. PMID- 15830183 TI - Islet isolation for allotransplantation: variables associated with successful islet yield and graft function. AB - AIMS/HYPOTHESIS: Efficient islet isolation is an important prerequisite for successful clinical islet transplantation. Although progressively improved, islet yield and quality are, however, unpredictable and variable and require standardisation. METHODS: Since 1989 we have processed 437 pancreases using the automated method. The donor characteristics, pancreas procurement, and digestion and purification procedures including a wide enzyme characterisation of these pancreases were analysed and correlated with islet yield and transplant outcome. RESULTS: By univariate analysis, islet yield was significantly associated with donor age (r=0.16; p=0.0009), BMI (r=0.19; p=0.0004), good pancreas condition (p=0.0031) and weight (r=0.15; p=0.0056), total collagenase activity (r=0.22; p=0.0001), adjusted collagenase activity/mg (r=0.18; p=0.0002), collagenase activity/solution volume (r=0.18; p=0.0002) and neutral protease activity/solution volume (r=0.14; p=0.0029). A statistically significant contribution to the variability of islet yield in a multivariate analysis performed on donor variables was found for donor BMI (p=0.0008). In a multivariate analysis performed on pancreas variables a contribution was found for pancreas weight (p=0.0064), and for a multivariate analysis performed on digestion variables we found a contribution for digestion time (p=0.0048) and total collagenase activity (p=0.0001). Twenty-four patients with type 1 diabetes received single islet preparations from single donors. In these patients, multivariate analyses showed that the reduction in insulin requirement was significantly associated with morphological aspects of islets (p=0.0010) and that 1-month C-peptide values were associated with islet purity (p=0.0071). CONCLUSIONS/INTERPRETATION: These data provide baseline donor, digestion and purification selection criteria for islet isolation using the automated method and indicate that the morphological aspect may be a clinically relevant measure of islets on which the decision for transplant can be based. PMID- 15830184 TI - Crosstalk between membrane potential and cytosolic Ca2+ concentration in beta cells from Sur1-/- mice. AB - AIMS/HYPOTHESIS: Islets or beta cells from Sur1(-/-) mice were used to determine whether changes in plasma membrane potential (V(m)) remain coupled to changes in cytosolic Ca(2+) ([Ca(2+)](i)) in the absence of K(ATP) channels and thus provide a triggering signal for insulin secretion. The study also sought to elucidate whether [Ca(2+)](i) influences oscillations in V(m) in sur1(-/-) beta cells. METHODS: Plasma membrane potential and ion currents were measured with microelectrodes and the patch-clamp technique. [Ca(2+)](i) was monitored with the fluorescent dye fura-2. Insulin secretion from isolated islets was determined by static incubations. RESULTS: Membrane depolarisation of Sur1(-/-) islets by arginine or increased extracellular K(+), elevated [Ca(2+)](i) and augmented insulin secretion. Oligomycin completely abolished glucose-stimulated insulin release from Sur1(-/-) islets. Oscillations in V(m) were influenced by [Ca(2+)](i) as follows: (1) elevation of extracellular Ca(2+) lengthened phases of membrane hyperpolarisation; (2) simulating a burst of action potentials induced a Ca(2+)-dependent outward current that was augmented by increased Ca(2+) influx through L-type Ca(2+) channels; (3) Ca(2+) depletion of intracellular stores by cyclopiazonic acid increased the burst frequency in Sur1(-/-) islets, elevating [Ca(2+)](i) and insulin secretion; (4) store depletion activated a Ca(2+) influx that was not inhibitable by the L-type Ca(2+) channel blocker D600. CONCLUSIONS/INTERPRETATION: Although V(m) is largely uncoupled from glucose metabolism in the absence of K(ATP) channels, increased electrical activity leads to elevations of [Ca(2+)](i) that are sufficient to stimulate insulin secretion. In Sur1(-/-) beta cells, [Ca(2+)](i) exerts feedback mechanisms on V(m) by activating a hyperpolarising outward current and by depolarising V(m) via store operated ion channels. PMID- 15830185 TI - Small intestinal enteropathy in non-obese diabetic mice fed a diet containing wheat. AB - AIMS/HYPOTHESIS: A deranged mucosal immune response and dietary factors may play an important role in the pathogenesis of type 1 diabetes. The aims of our work were to look for the presence of small intestinal enteropathy in non-obese diabetic (NOD) mice in relation to the presence of wheat proteins in the diet, and to assess their role in the risk of developing diabetes. METHODS: Female NOD mice were fed a standard or gluten-free diet or a gluten-free diet with the addition of wheat proteins (MGFD). Small intestine architecture, intraepithelial CD3(+) infiltration, epithelial expression of H2-IA, mRNA for IFN-gamma and IL-4 were assessed. RESULTS: NOD mice fed a standard diet showed reduced villous height, increased intraepithelial infiltration by CD3(+) cells and enhanced expression of H2-IA and IFN-gamma mRNA when compared with mice on the gluten-free diet. The cumulative diabetes incidence at 43 weeks of age was 65% in the latter and 97% in the former (p<0.01). Mice on MGFD also showed increased epithelial infiltration and a higher incidence of diabetes. CONCLUSIONS/INTERPRETATION: Mice fed a wheat-containing diet showed a higher incidence of diabetes, signs of small intestinal enteropathy and higher mucosal levels of proinflammatory cytokines. PMID- 15830186 TI - Association between plasma activities of semicarbazide-sensitive amine oxidase and angiotensin-converting enzyme in patients with type 1 diabetes mellitus. AB - AIMS/HYPOTHESIS: Plasma semicarbazide-sensitive amine oxidase (SSAO) is elevated in patients with type 1 and type 2 diabetes and has been implicated in the pathophysiology of diabetic late complications. The regulation of SSAO production remains unknown. We studied correlations between plasma SSAO activity and parameters associated with diabetic late complications. METHODS: Plasma SSAO was measured in a well-characterised group of 287 patients with type 1 diabetes. Standard statistical methods were used to investigate correlations with clinical parameters and components of the renin-angiotensin system. RESULTS: Overall, plasma SSAO was elevated, at 693+/-196 mU/l (mean+/-SD; normal controls 352+/-102 mU/l). Plasma SSAO was higher in the group with late complications or hypertension, and in patients treated with ACE-inhibitors. In univariate analysis a significant positive correlation (p<0.001, r=0.27) was found between plasma SSAO and serum ACE activity in patients untreated with ACE inhibitors or angiotensin II receptor antagonists (n=221), but plasma SSAO did not differ by ACE I/D genotype. Plasma SSAO correlated positively with duration of diabetes, HbA(1)c and plasma renin, and negatively with plasma angiotensinogen and body mass index. A multiple regression analysis including these variables resulted in serum ACE activity (p<0.001), ACE genotype (negatively, p<0.001) and HbA(1)c (p=0.023) as explaining variables. CONCLUSIONS/INTERPRETATION: Results suggest that a common factor is involved in the regulation of both plasma SSAO and serum ACE, which is different from the genetic determination of ACE activity. PMID- 15830187 TI - Insulin resistance induced by sucrose feeding in rats is due to an impairment of the hepatic parasympathetic nerves. AB - AIMS/HYPOTHESIS: A considerable proportion of whole-body insulin-stimulated glucose uptake is dependent upon the hepatic insulin-sensitising substance (HISS) in a pathway mediated by the hepatic parasympathetic nerves (HPNs). We tested the hypothesis that a high-sucrose diet leads to the impairment of the HPN-dependent component of insulin action. METHODS: We quantified insulin sensitivity using the rapid insulin sensitivity test, a modified euglycaemic clamp. Quantification of the HPN-dependent component was achieved by administration of a muscarinic receptor antagonist (atropine, 3 mg/kg). RESULTS: Insulin sensitivity was higher in standard-fed than in sucrose-fed Wistar rats (305.6+/-34.1 vs 193.9+/-13.7 mg glucose/kg body weight; p<0.005) and Sprague-Dawley rats (196.4+/-5.9 vs 95.5+/ 16.3 mg glucose/kg body weight; p<0.01). The HPN-independent component was similar in the two diet groups. Insulin resistance was entirely due to an impairment of the HPN-dependent component in both Wistar rats (164.3+/-28.1 [standard-fed] vs 26.5+/-7.5 [sucrose-fed] mg glucose/kg body weight; p<0.0001) and Sprague-Dawley rats (111.7+/-9.5 vs 35.3+/-21.4 mg glucose/kg body weight; p<0.01). Furthermore, HPN-dependent insulin resistance in Sprague-Dawley rats was already evident after 2 weeks of a high-sucrose diet (28.5+/-7.6 [2 weeks], 35.3+/-21.4 [6 weeks], 17.9+/-5.4 [9 weeks] mg glucose/kg body weight) and was independent of the nature of sucrose supplementation (12.3+/-4.7 [solid] and 17.9+/-5.4 [liquid] mg glucose/kg body weight). CONCLUSIONS/INTERPRETATION: Our results support the hypothesis that insulin resistance caused by sucrose feeding is due to an impairment of the HPN-dependent component of insulin action, leading to a dysfunction of the HISS pathway. PMID- 15830189 TI - Two of the three groEL homologues in Rhizobium leguminosarum are dispensable for normal growth. AB - Although many bacteria contain only a single groE operon encoding the essential chaperones GroES and GroEL, examples of bacteria containing more than one groE operon are common. The root-nodulating bacterium Rhizobium leguminosarum contains at least three operons encoding homologues to Escherichia coli GroEL, referred to as Cpn60.1, Cpn60.2 and Cpn60.3, respectively. We report here a detailed analysis of the requirement for and relative levels of these three proteins. Cpn60.1 is present at higher levels than Cpn60.2, and Cpn60.3 protein could not be detected under any conditions although the cpn60.3 gene is transcribed under anaerobic conditions. Insertion mutations could not be constructed in cpn60.1 unless a complementing copy was present, showing that this gene is essential for growth under the conditions used here. Both cpn60.2 and cpn60.3 could be inactivated with no loss of viability, and a double cpn60.2 cpn60.3 mutant was also constructed which was fully viable. Thus only Cpn60.1 is required for growth of this organism. PMID- 15830188 TI - Synergistic effect of portal glucose and glucagon-like peptide-1 to lower systemic glucose and stimulate counter-regulatory hormones. AB - AIMS: Glucagon-like peptide-1 (GLP-1) is an insulinotropic hormone released from the gut in response to nutrients. Besides its well-established direct effect on pancreatic beta cells, GLP-1 may also act by activating sensors in the hepatoportal area. We therefore studied the impact of putative GLP-1 sensors in the splanchnic circulation. METHODS: We infused GLP-1 into the portal vein of conscious dogs, while also infusing glucose intraportally or systemically. In the first experiment, we infused glucose intraportally, simulating portal glucose values obtained during a previous mixed-meal test, with or without co-infusion of intraportal GLP-1. In the second experiment, by infusing glucose systemically, with or without intraportal GLP-1, we investigated whether the effects of systemic glucose with or without portal GLP-1 infusion are similar to those observed in the first experiment. RESULTS: Intraportal infusion of GLP-1 and glucose significantly raised peripheral GLP-1 levels, but did not produce an insulin response different from intraportal glucose alone. However, the resulting peripheral glycaemia was significantly lower compared to glucose infusion alone, and there were elevations in glucagon, cortisol and lactate. In contrast to the portal glucose infusions, there were no significant differences in glucose, insulin, glucagon, cortisol or lactate levels between systemic glucose infusion with or without GLP-1. CONCLUSIONS/INTERPRETATION: Portal GLP-1 and portal glucose, but not systemic glucose, can produce decreased peripheral glucose levels independently of hyperinsulinaemia. This suggests that portal GLP-1 and glucose receptors mediate insulin-independent changes in peripheral glycaemia and determine a strong counter-regulatory response, as reflected by changes in glucagon and cortisol. PMID- 15830190 TI - Determination of ethyl carbamate in alcoholic beverages: an interlaboratory study to compare HPLC-FLD with GC-MS methods. AB - An international interlaboratory study on the determination of ethyl carbamate in alcoholic beverages by a new HPLC-FLD and by the official GC-MS methods is presented. The aim of this study was to improve the knowledge about precision and accuracy parameters of the new method and to compare the performance of both HPLC and GC methods. Five different samples representing table wines, fortified wines (red and white), distilled spirits, and wine spirits were available for analysis by each participant. Despite the low number of participants (6), the results obtained by the laboratories using the HPLC-FLD method are comparable to those obtained by GC-MS methods. The present study emphasizes the possibility to use, as routine, a much simpler analytical method than the current reference method by GC-MS for ethyl carbamate determination in alcoholic beverages. PMID- 15830191 TI - Rapid voltammetric determination of nitroaromatic explosives at electrochemically activated carbon-fibre electrodes. AB - The electrochemical behaviour of some nitroaromatic explosives (2,4,6 trinitrotoluene, TNT; 2,6-dinitrotoluene, 2,6-DNT; 2-nitrotoluene, 2-NT; 2-amino 4,6-dinitrotoluene, 2-A-4,6-DNT; 3,5-dinitroaniline, 3,5-DNA; and nitrobenzene, NB) at electrochemically activated carbon-fibre microelectrodes is reported. Electrochemical activation of such electrode material by repeated square-wave (SW) voltammetric scans between 0.0 and +2.6 V versus Ag/AgCl, produced a dramatic increase in the cathodic response from these compounds. This is attributed to the increase of the carbon-fibre surface area, because of its fracture, and the appearance of deep fissures along the main fibre axis into which the nitroaromatic compounds penetrate. Based on the important contribution of adsorption and/or thin layer electrolysis to the total voltammetric response, a SW voltammetric method for rapid detection of nitroaromatic explosives was developed. No interference was found from compounds such as hydrazine, phenolic compounds, carbamates, triazines or surfactants. The limits of detection obtained are approximately 0.03 microg mL(-1) for all the nitroaromatic compounds tested. The method was applied for the determination of TNT in water and soil spiked samples; recoveries were higher than 95% in all cases. PMID- 15830192 TI - Assessment of the essential element and heavy metal content of edible fish muscle. AB - The aim of this work was to determine the concentrations of some essential and toxic elements in the muscle of ten species of commercial fish consumed in Portugal. We combined two different techniques for determination of the elements- energy-dispersive X-ray fluorescence (EDXRF) was used to quantify K, Ca, Fe, Zn, Se, Rb, and Sr and flame atomic-absorption spectrometry for analysis of Cr, Ni, Cu, Cd, Hg, and Pb. The latter technique was used because of its higher sensitivity, because these elements were not detected by EDXRF. The results obtained show a similar pattern for the trace elements. K and Ca are present at the highest concentrations in all the samples studied, from 0.6-1.3% and from 0.04-0.08%, respectively, followed by Zn, Fe, Sr, Se, and Rb. Sr is present at higher concentrations than Rb in all the species studied except meagre. Concentrations of the elements in octopus do not follow this pattern--Fe is present at a higher concentration than Zn. Low concentrations of Cr (0.66-1.5 microg g(-1)), Ni (0.11-0.24 microg g(-1)), Cd (0.01-0.08 microg g(-1)), Hg (0.49 2.74 microg g(-1)), and Pb (0.02-0.06 microg g(-1)) were observed in all the samples analysed. The concentration of Hg was highest in Helicolenus dactylopterus--5.4 microg g(-1) in one sample. PMID- 15830193 TI - Spatial object representation and its use in planning eye movements. AB - The eye movements we make to look at objects require that the spatial information contained in the object's image on the retina be used to generate a motor command. This process is known as sensorimotor transformation and has been generally addressed using simple point targets. Here, we investigate the sensorimotor transformation involved in planning double saccade sequences directed at one or two objects. Using both visually guided saccades toward stationary objects and objects subjected to intrasaccadic displacements, and memory-guided saccades, we found that the coordinate transformations required to program the second saccade were different for saccades aimed at a new target object and saccades that scanned the same object. While saccades aimed at a new object were updated on the basis of the actual eye position, those that scanned the same object were performed with a fixed amplitude, irrespective of the actual eye position. Our findings demonstrate that different abstract representations of space are used in sensory-to-motor transformations, depending on what action is planned on the objects. PMID- 15830194 TI - Elevated lactate as an early marker of brain injury in inflicted traumatic brain injury. AB - BACKGROUND: Traumatic brain injury is a major cause of disability and death in the pediatric population. The metabolic and neurochemical abnormalities that underlie traumatic brain injury remain poorly understood, but hypoxia-ischemic injury might play an important role. OBJECTIVE: This study evaluated children with inflicted traumatic brain injury using magnetic resonance spectroscopy (MRS). We postulated that children with hypoxic-ischemic injury indicated by elevated lactate in the acute phase of injury will have worse early neurological status and short-term clinical outcomes than those without lactate upon MRS. MATERIALS AND METHODS: This prospective study employed proton MRS to sample bilaterally the frontal lobes and the parasagittal cortex within the parietal and occipital lobes of 11 patients with inflicted traumatic brain injury who were undergoing a clinical MRI examination. Patients' measured clinical course while hospitalized included initial neurological evaluation, presence of seizure activity, need for admission to the pediatric intensive care unit (PICU), number of days hospitalized, presence of retinal hemorrhages and presence of bone fractures. Measurement of outcome was determined using the Pediatric Overall Performance Category Scale (POPCS; 1=good performance; 6=death). RESULTS: Four children demonstrated elevated lactate and diminished N-acetyl aspartate (a neuronal marker) within several regions, indicating global ischemic injury (lactate-positive global group). These four children all had seizure activity and abnormal initial neurological examinations and required admission to the PICU. The mean POPCS for this group was 3.25. In four other children, lactate was detected within at least one region, indicating a focal ischemic injury (lactate positive focal group); two of these children had seizure activity, and two had an abnormal initial neurological examination. The mean POPCS score was 1.5 for this group. The remaining three children had no evidence of lactate upon MRS (lactate negative group). These children did not have seizure activity, did not require admission to the PICU, nor did they have initial abnormal neurological examinations. The mean POPCS score was 1.3 for this group. SUMMARY: Patients with inflicted traumatic brain injury and evidence of hypoxic-ischemic injury as indicated by elevated lactate on MRS tend to have worse early neurological status and early outcome scores. Lactate levels as sampled by MRS might predict early clinical outcome in inflicted traumatic brain injury. PMID- 15830195 TI - The potential of Pseudozyma yeastlike epiphytes for the production of heterologous recombinant proteins. AB - Although Basidiomycetes represent the most evolved class of fungi, they have been neglected with regard to recombinant gene expression. In this work, basidiomycetous yeasts belonging to Pseudozyma spp. were studied with respect to their amenability to heterologous protein production. Single plasmid or cotransformation experiments routinely afforded 100 to 200 independent transformants for the two tested species of Pseudozyma. Green fluorescent protein (GFP) was expressed in the correctly folded conformation, as demonstrated by fluorescence microscopy, and hen egg white lysozyme (HEWL) was expressed in its active form, as revealed by its lytic activity on Micrococcus lysodeikticus cells. Protease analysis established that Pseudozyma spp. contained equivalent or less extracellular protease activity than yeasts and far less protease activity than ascomycetous filamentous fungi in similar culture conditions. This proteolytic activity was inhibited by over 97% with a combination of PMSF and Pepstatin A. N-glycosylation patterns of native Pseudozyma flocculosa secreted proteins were comprised of one or a few short glycan chains that possess a classic eukaryotic structure typical of higher fungi and animal cells. This is the first report of a Basidiomycete that possesses multiple intrinsic characteristics necessary for use as a heterologous gene expression system. PMID- 15830196 TI - Stable genetic transformation of Larix gmelinii L. by particle bombardment of zygotic embryos. AB - We report a new protocol for the stable transformation of Larix gmelinii. Thirty mature zygotic embryos precultured for 3 days on solid medium supplemented with benzyladenine were bombarded with plasmids pUC-GHG (GUS, HPT, and GFP genes) or pBI221-HPT (HPT and GUS genes). After a 2-month culture on selection medium, hygromycin-resistant calli appeared on the surfaces of the necrotic embryos. The frequencies of embryos with resistant calli were 18.4% and 17.4% in the transformations with pUC-GHG and pBI221-HPT DNA, respectively. More than 20 adventitious shoots formed from each of the transgenic calli. Of 17 elongated shoots selected for culturing on a rooting medium, five shoots rooted after 2 months. Expression of the GFP and GUS genes was detected in the resistant tissues by microscopic observations and by a histological GUS activity assay, respectively. PCR and Southern analysis confirmed the stable insertion of the introduced DNA into the genome. PMID- 15830197 TI - Percutaneous laser-assisted recanalization of long chronic iliac artery occlusions: primary and mid-term results. AB - We report the primary and mid-term outcome of patients with long chronic iliac artery occlusions after percutaneous excimer-laser-assisted interventional recanalization. Between 2000 and 2001, 43 patients with 46 chronic occlusions of either the common iliac artery (n=27), the external iliac artery ( n=13) or both (n=3) underwent laser-assisted percutaneous transluminal angioplasty and implantation of stents. The average length of the occlusion was 57.1+/-26 mm. After laser-assisted angioplasty and implantation of a total of 60 stents, the patients were followed up for up to 4 years. Patency rates were analyzed by ankle brachial index (ABI) measurement and duplex ultrasound. The primary technical success rate was 95.3%, with a major complication rate of 6.9%. Clinical improvement as categorized by the Rutherford guidelines could be observed in 97.6% of cases. The ABI of all patients improved from an average of 0.46+/-0.08 before intervention to 0.97+/-0.13 at the end of the follow-up period. The overall primary patency rate was 86.1%. Four reinterventions were successful (secondary patency rate 95.4%). The mid-term results of the percutaneous recanalization of iliac artery occlusions with primary and secondary patency rates of 86.1 and 95.4% are similar to those of the treatment of short stenoses. PMID- 15830198 TI - [Functional results with two multifocal intraocular lenses (MIOL). Array SA40 versus Acri.Twin]. AB - BACKGROUND: A multifocal intraocular lens (MIOL) with diffractive and aspherical optical design and asymmetrical light distribution (Acri. Twin, Acri.Tec) was compared to the standard MIOL, the Array SA40. PATIENTS AND METHODS: After 6 months, 16 patients with bilateral implantation of the Acri. Twin and 14 patients with bilateral Array IOL were examined to assess uncorrected and best corrected visual acuity for distance and near, contrast sensitivity under photopic and mesopic conditions, stereopsis, pupil diameter, and centration of the IOL. RESULTS: A significant difference was observed between the two MIOL in improved near visual acuity of the Acri. Twin MIOL (0.8 vs 0.4). Most of the patients described dysphotic phenomena, which were somewhat different for both MIOL, but expressed general satisfaction except for one patient. Concerning contrast sensitivity both MIOL were significantly inferior to data published for an aspherical monofocal IOL. PMID- 15830200 TI - [Diurnal frequency patterns of errors made in ophthalmo-surgery. A clinical analysis related to evidence based medicine]. AB - BACKGROUND AND OBJECTIVES: Three factors are usually considered in determining optimal patient care, the goal of evidence based medicine (EBM): (1) approximation to the best medical knowledge, (2) clinical competence of the doctor and self reflection, (3) taking into account the individual values and beliefs of the patient. Must the limited loading capacity of the doctor also be considered as the fourth factor in ophthalmic surgery? METHODS: Follow-up examination of 8,212 cataract operation reports was carried out. We recorded complications (active errors) and specific intra-operative features (latent errors). RESULTS: The errors were not distributed evenly between 07.45 h and 13.14 h (chi(2)-test: P<0.001). They reach a maximum around 11 h. CONCLUSIONS: Usual recommendations by EBM claim a surgeon to be a machine. They should, however, take into account systematic changes in efficiency that can probably be reduced by simple nutritive interventions. PMID- 15830199 TI - [Functional significance of adenosine receptors in the eye and their dysregulation in pseudoexfoliation syndrome]. AB - BACKGROUND: Adenosine regulates many physiologic processes, such as aqueous humor secretion and intraocular pressure, via activation of its receptors. We investigated the expression of the receptor isoforms in eyes with PEX syndrome, which is associated with anterior chamber hypoxia and elevated intraocular pressure. MATERIALS AND METHODS: The mRNA and protein expression of the adenosine receptor subtypes in anterior segment tissues of patients with PEX syndrome, glaucomas, and normal control eyes were analyzed comparatively. RESULTS: Whereas the receptor subtypes A1, A2a, and A2b displayed no differential expression between PEX and control tissues, expression of the A3 adenosine receptor was consistently enhanced in the nonpigmented ciliary epithelium of all PEX eyes, independent of the presence of glaucoma, compared to normal and glaucomatous control eyes. CONCLUSION: Considering the known role of the A3 adenosine receptor in modulating aqueous humor secretion, its selective upregulation in the ciliary epithelium may confer cytoprotection and be accessible to therapeutic intervention in PEX patients. PMID- 15830201 TI - [Anatomical and functional results after elective macular surgery]. AB - PURPOSE: The purpose of this paper is to assess the anatomical and functional results after macular surgery in a large group of patients. METHODS: Between June 1995 and December 2001, 381 eyes underwent vitreous surgery for macular pucker (n=244) or macular holes (n=137) with a standard pars plana vitrectomy (PPV) with induction of posterior vitreous separation, membrane peeling, peeling of the internal limiting membrane (no ICG staining was used), and gas instillation (SF(6)). RESULTS: A second surgical intervention due to vision-threatening complications after PPV had to be performed in 8 of 381=2.1%. In the macular pucker group, metamorphopsias improved in 46.6% and the median of visual acuity (VA) improved from preoperative 0.3 to postoperative 0.5. Hole closure of macular holes was obtained in 92.2%; the median of VA improved in this group from preoperative 0.2 to postoperative 0.4. CONCLUSION: In our group a second vitreoretinal procedure due to vision-threatening complications had to be performed in 2.1%. Compared to the spontaneous course, PPV for macular pucker or macular hole has a very positive influence on functional parameters. PMID- 15830202 TI - Miller-Fisher syndrome mimicking intracranial hypertension following head trauma. AB - INTRODUCTION: Miller-Fisher syndrome (MFS) is a polyneuropathy with benign outcome characterized by ophthalmoplegia, limb ataxia and tendon areflexia. Impaired consciousness level and intracranial hypertension are very rare symptoms in MFS. CASE REPORT: We describe the case of a 5-year-old girl who showed intracranial hypertension, transient coma and respiratory failure after mild head injury; moreover the patient showed mild ataxia, areflexia, ophthalmoplegia and autonomic disturbances. These symptoms were suggestive of MFS. Electrophysiologic studies and laboratory tests confirmed the diagnosis and immunoglobulins and steroids were given. The child showed a progressive clinical improvement and the final outcome was good. CONCLUSION: This case, initially managed as trauma injury due to the presence of suggestive signs and clinical history, maskered an atypical presentation of Miller-Fisher syndrome, a rare disorder of central nervous system. PMID- 15830203 TI - Haemorrhagic shock caused by splenic rupture following routine colonoscopy. PMID- 15830204 TI - Outcome of overlapping anal sphincter repair after 3 months and after a mean of 80 months. AB - BACKGROUND: The aim of this study was to determine the functional results of anal sphincter repair after a long follow-up (mean 80.1 months). METHODS: In the period 1990-2002, 65 sphincter repairs were performed. Obstetric trauma was the cause of incontinence in 72.3% cases, fistulotomy in 13.8%, nonspecific trauma in 9.2%, and war injury in 4.6%. At the time of surgery, 12 patients had undergone an urgent stoma procedure. In all cases, anal manometry, electromyography, and defecography were performed. The severity of incontinence was evaluated preoperatively using the Wexner score system. Anterior sphincteroplasty was performed in 52 cases, lateral in 9 cases, and posterior in 4 cases. RESULTS: The results were determined according to the Wexner score system and the Browning Parks scale. The Wexner score was calculated 3 months after operation and during every follow-up visit. Preoperative scores and those at the first and last follow up visits were analyzed. Three months after operation excellent results were achieved in 55.5%, good in 18.5%, fair in 16.9%, and poor in 9.2% patients. After follow-up (mean 80.1 months), 26.8% had excellent results, 21.4% had good results, 12.5% had fair results, and 39.3% of patients had a poor outcome. Results determined by the Wexner score system improved from 17.8 preoperatively to 3.6 three months after operation, but deteriorated over time to 6.3 after longer follow-up (p<0.001). CONCLUSION: Overlapping sphincter repair provides satisfactory results in more than two-thirds of patients initially, but the results tend to worsen over time and are satisfactory in half of patients after longer follow-up. PMID- 15830205 TI - Biokinetic modeling of uranium in man after injection and ingestion. AB - Uranium is a naturally occurring primordial radioactive element. Small amounts found in air, water, and food are regularly consumed and inhaled by humans. Even the military, medical, and industrial use of depleted uranium can affect humans. There is an appreciable retention of incorporated uranium in skeleton, kidneys, and liver, and a review of respective effective dose coefficients has been given by the International Commission on Radiological Protection (ICRP) in its "Publication 69"; however, data regarding retention in organs or tissues and rates of urinary and fecal excretion for different age groups are incomplete. Therefore, the present study provides retention data that have been calculated for uranium in all compartments and for urinary and fecal excretion, following acute and chronic injection and ingestion for six age groups. The calculations are based on the current ICRP biokinetic model for uranium, and the data can be plotted by using any mathematical software to obtain the retention data at any time after incorporation or to calculate the internal average organ dose induced by uranium provided that specific absorbed fractions are available. The dynamic relationship of the retention in plasma and blood after intravenously and orally administered uranium can easily be derived from the database for injection and ingestion. The calculated contents of uranium in organs or tissues (using the uranium concentration in foodstuffs published by UNSCEAR for Europeans) are compared with autopsy data available in the literature. According to this model, the whole body of a 75-year-old man contains 7 microg uranium, of which 76% is in the skeleton, 1% in the kidneys, and 2.1% in the liver. PMID- 15830207 TI - Sugar-mediated transcriptional regulation of the Gap gene system and concerted photosystem II functional modulation in the microalga Scenedesmus vacuolatus. AB - Partial cDNAs corresponding to the GapA, GapC and GapN genes that encode the three different glyceraldehyde-3-phosphate dehydrogenases (GAPDHs) of the green microalga Scenedesmus vacuolatus SAG 211-8b have been cloned and characterized. Northern blot experiments, as well as immunoblots and activity measurements, demonstrate a differential regulation by sugars of the components of the algal Gap gene system. Addition of glucose or other metabolizable sugars to photoautotrophic cultures promoted a drastic repression of the GapA gene and depletion to negligible levels of the corresponding GAPDHA, a chloroplastic protein involved in photosynthetic CO2 assimilation. By contrast, expression of the GapC and GapN genes encoding their cytosolic counterparts involved in glycolysis was enhanced. However, no down-regulation of the GapA gene by glucose took place in the dark, indicating that the observed effect is associated with sugar assimilation in the light. Likewise, glucose promoted in illuminated algal cultures a severe decrease of photosystem II functionality, estimated by O2 evolution activity, thermoluminescence emission and D1 protein level, while again, no effect was observed in the dark. On the basis of the correlation found between photosystem II performance and sugar transcriptional regulation of the GapA gene, a scenario of sugar-mediated regulation of photosynthetic metabolism in microalgae is proposed that will help to explain the so-called glucose bleaching effect in photosynthetic eukaryotes. PMID- 15830206 TI - Human DNA topoisomerase I: relaxation, roles, and damage control. AB - Human DNA topoisomerase I is an essential enzyme involved in resolving the torsional stress associated with DNA replication, transcription, and chromatin condensation. The catalytic cycle of the enzyme consists of DNA cleavage to form a covalent enzyme-DNA intermediate, DNA relaxation, and finally, re-ligation of the phosphate backbone to restore the continuity of the DNA. Structure/function studies have elucidated a flexible enzyme that relaxes DNA through coordinated, controlled movements of distinct enzyme domains. The cellular roles of topoisomerase I are apparent throughout the nucleus, but the concentration of processes acting on ribosomal DNA results in topoisomerase I accumulation in the nucleolus. Although the activity of topoisomerase I is required in these processes, the enzyme can also have a deleterious effect on cells. In the event that the final re-ligation step of the reaction cycle is prevented, the covalent topoisomerase I-DNA intermediate becomes a toxic DNA lesion that must be repaired. The complexities of the relaxation reaction, the cellular roles, and the pathways that must exist to repair topoisomerase I-mediated DNA damage highlight the importance of continued study of this essential enzyme. PMID- 15830208 TI - Molecular and biochemical characterization of a protein cyclophilin from the nematode Haemonchus contortus( P ). AB - We have cloned, sequenced and expressed a gene of Haemonchus contortus that encodes a protein (termed HcCYP) consisting of a cyclophilin domain and an RNA recognition motif (RRM). An antiserum raised against the recombinant protein showed that HcCYP was present in the insoluble fraction (mostly nuclear) of the parasite homogenate. The recombinant protein possessed the typical cis-trans peptidyl-prolyl isomerase activity of cyclophilins and this activity was inhibited by the immunosuppressant cyclosporin A. The N-terminal portion of the molecule, carrying the RRM, was able to bind to nucleic acids, whereas the C terminal portion did not have any binding activity. The possible function of HcCYP in the parasite is discussed on the basis of information available on similar proteins in other organisms. PMID- 15830210 TI - Genetic mosaics in the massive persisting rhizosphere colony "shiro" of the ectomycorrhizal basidiomycete Tricholoma matsutake. AB - The ectomycorrhizal basidiomycete Tricholoma matsutake produces commercially valuable fruit bodies "matsutake" on a massive persisting rhizosphere aggregate of mycelia and mycorrhizas called "shiro." Using inter-retrotransposon amplified polymorphism analysis, we attempted to explore the potential diversity within the population of T. matsutake isolated from small Pinus densiflora woodlands located in various parts of Japan. In general, random phylogenetic relationship was noted among T. matsutake tested. The population from each limited sampling area was highly heterogeneous. Even some isolates from fruit bodies produced in the same shiro and those from spores in the same fruit bodies were found to be genetically diverse, indicating the occurrence of genetic mosaics in shiro. In a mosaic shiro, heterologous genets produced their fruit bodies concurrently. Data suggested that the dispersal of spores through sexual reproduction may have been more prevalent than generally accepted in T. matsutake to bring mosaicism and coordination of heterologous genets within the shiro. Implementation of management taking such diversity into consideration is urgently needed for the restoration of devastated matsutake fields in Japan. Exploration of individual clones in mosaic fungal resources that promote colonization and fruit body production is necessary for it. PMID- 15830211 TI - Hyphal growth and mycorrhiza formation by the arbuscular mycorrhizal fungus Glomus claroideum BEG 23 is stimulated by humic substances. AB - Effects of humic substances (humic acid or fulvic soil extract) or saprophytic microorganisms (Paecilomyces lilacinus and an unidentified actinomycete) on growth of mycelium and mycorrhiza formation by Glomus claroideum BEG23 were studied in a hydroponic system. Humic substances stimulated root colonization and production of extraradical mycelium by the mycorrhizal fungus. Both humic and fulvic acids tended to decrease populations of culturable bacteria and fungi in the cultivation system, indicating a moderately antibiotic activity. The addition of saprophytic microorganisms able to use humic substances to the cultivation system further stimulated the development of the mycorrhizal fungus. However, stimulation of G. claroideum was also observed when the saprophytic microorganisms were heat-killed, suggesting that their effect was not linked to a specific action on humic substances. The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi. PMID- 15830212 TI - Pull-out strength of the suprapedicle claw construct: a biomechanical study. AB - The pull-out of the superior screw is a well recognized problem in anterior instrumentation of the spine for scoliosis. A biomechanical pull-out study of anterior vertebral body screw in cadaveric thoracic spine was therefore designed to investigate and compare the pull-out strength of three different anterior vertebral body fixations using the AO Universal Spine System: simple bicortical screw, bicortical screw with an opposite washer (sometimes called pull-out resistant nut), and a new construct made of a bicortical screw with the addition of a suprapedicular hook on the same vertebra (or claw construct). The T4 to T9 vertebral bodies from six human cadavers (total of 36 specimens) were instrumented with three different instrumentation constructs after measuring the bone mineral density of each individual vertebra. After stabilization of the vertebral bodies, the screws were extracted employing a material testing system using axial pull-out. The maximum axial forces were recorded at the time of the construct failure. The mean ultimate fixation strength (UFS) values after being adjusted for bone mineral density and vertebral body diameter were 631, 711, and 1244 N for the three different constructs, respectively (screw alone, screw with an opposite washer, and screw with a suprapedicle claw). The difference in UFS was not significant for the first two constructs tested (screw alone and screw with an opposite washer). However, the difference in ultimate fixation strength between the claw and the other constructs was highly significant (P<0.0001). Specimens with low BMD did not benefit as much from claw construct as the ones did with a normal BMD. The failure mode of each construct was described, but was in neither case judged dangerous for the spinal cord. This study shows that the suprapedicle claw construct improves the pull-out strength of an anterior vertebral body screw by 80%, and changes the mode of failure so as not to rely only on the screw characteristics or solely on the vertebral body. By adding a suprapedicle hook in a claw configuration, one may prevent superior screw pull out in anterior spine surgery for scoliosis. PMID- 15830213 TI - The suprapedicle claw construct in anterior scoliosis surgery. AB - Proximal screw pullout is a well-recognized problem in anterior scoliosis surgery, with a rate of pseudarthrosis or screw pullout ranging from 15 to 30%. To prevent screw pullout at the top of the construct, the authors have devised the concept of a claw for the top instrumented vertebra. The claw consists of a classic anterior vertebral body screw inserted parallel to the inferior end-plate and in the posterior portion of the vertebral body 8 mm in front of the spine canal. After rib desarticulation, a laminar hook of a small size is inserted over the superior aspect of the pedicle of the same vertebra. The rod is then inserted into the two side openings of the screw and the hook. Compression across the hook and the screw is then performed, making a claw construct. This concept can also be extended in the case of early revision for a proximal screw pullout, where it is possible to revise the instrumentation with an offset connector linking the rod to the superior portion of the pedicle where the suprapedicule hook has been inserted. We report two cases where a suprapedicle claw was successfully used in anterior scoliosis correction of a right thoracic curve. Such a concept may represent the solution to proximal screw pullout in anterior scoliosis correction. PMID- 15830214 TI - A prospective study of the interrelationship between subjective and objective measures of disability before and 2 months after lumbar decompression surgery for disc herniation. AB - The value of range of motion (ROM) as an indicator of impairment associated with spinal problems, and in monitoring changes in response to treatment, is a controversial issue. The aim of this study was to examine the interrelationship between subjective disability (Roland-Morris scores) and objectively measured impairment (ROM), both before and in response to spinal decompression surgery, in an older group of patients with herniated lumbar disc (DH). Seventy-six individuals took part in the study: 33 patients (mean age 57 years, SD 9 years) presenting with DH and for whom decompression surgery was planned, and 43 controls (mean age 57 years, SD 7 years), with no history of back pain requiring medical treatment. In the patient group, pain intensity (leg and back; visual analog score), self-rated disability (Roland-Morris score), certain psychological attributes, and ROM of the spine (Spinal Mouse) were measured before and 2 months after decompression surgery. In addition, the patients rated the success of surgery on a 1-5 Likert scale. The pain-free control group performed only the tests of spinal mobility. Before surgery, compared with matched controls, significantly lower values were observed in the DH patients for standing lumbar lordosis (p=0.01), and for range of flexion of the lumbar spine (ROF(lumbar)) (p=0.0006), but not of the hips (ROF(hip)) (p=0.14). Roland-Morris Disability scores correlated significantly with ROF(lumbar) (r=0.61, p=0.0002), but less well with ROF(hip)(r=0.43, p=0.01). Two months after surgery, there were significant reductions in back pain and leg pain (p=0.0001) and in Roland-Morris Disability scores (p=0.019). There was also a significant decrease in the group mean values for lumbar lordosis angle (i.e., a "flatter" spine after surgery, p=0.002) and ROF(lumbar) (p=0.038). ROF(hip) showed a (nonsignificant) tendency to increase (p=0.08) towards normal control values. As a result of these two opposing changes, the range of total trunk flexion showed no significant changes from pre-surgery to 2 months post-surgery (p=0.60). On an individual basis, there was a highly significant relationship between the change in self-rated disability scores and the change in ROF(lumbar), pre-surgery- to 2 months post-surgery (r= 0.82; p<0.0001). Changes in ROF(hip) showed no such relationship (r= -0.30, p=0.10). The patients in the "poor" outcome group ("surgery didn't help"; 9%) had a significantly greater reduction in ROF(lumbar) post-surgery compared with the "good" outcome group ("surgery helped"; 91%) (p=0.04). In stepwise linear regression, the change in ROF(lumbar) was the only variable accounting for the change in self-rated disability pre-surgery to post-surgery (variables not included: pain intensity, psychological factors). The pivotal role of lumbar mobility in explaining disability emphasizes the importance of measuring lumbar and hip ranges of motion separately, as opposed to "global trunk motion." In the patient group examined, the determination of lumbar spinal mobility provides a valid, objective measure of function, that shows differences from normal matched controls, that correlates well with self-rated disability, and the changes in which correlate extremely well with subjective changes in disability following surgery. PMID- 15830215 TI - Scheuermann's kyphosis: surgical management. AB - Indications for surgery in Scheuermann disease are not well codified and remain rare, as the natural history of the disease is in most cases benign. In the immature adolescent, conservative treatment, such as bracing or casting, can be tried for moderate curves. For larger curves, or in the adult, conservative treatment is usually not effective, and surgery can be considered. Such indications are mostly cosmetic for large curves above 75 degrees. Pain over the deformity or in the low back may represent another surgical indication, especially in the adult group. The question of anterior release or straight posterior fusion has become more of an actuality with the advent of powerful, third-generation stiff segmental instrumentation. However, the long-term results of a modern, posterior-only instrumentation fusion are not known. Concern about loss of correction, late pseudarthrosis or the need to remove instrumentation for infected hardware or due to late pain at the operative site must make us careful about choosing this method. Very rigid and large curves still require an anterior release, either done in a conventional or mini-open fashion, or through video assisted thoracoscopic surgery. The extent of the posterior instrumentation has now been better defined. One must fuse the whole Cobb angle without hypercorrection and stop distally, above the first lordotic disc, to avoid sagittal decompensation. New approaches such as short anterior fusion with bone on-bone techniques and pedicle substraction osteotomies have not yet been reported in the literature as having been used for treating Scheuermann's kyphosis. These should be considered experimental. PMID- 15830216 TI - Dimanganese catalase--spectroscopic parameters from broken-symmetry density functional theory of the superoxidized Mn(III)/Mn(IV) state. AB - Broken-symmetry density functional theory was used to study the catalytic center of manganese catalase in the superoxidized Mn(III)/Mn(IV) state. Heisenberg exchange coupling constants, 55Mn and 14N hyperfine coupling constants (hfcs) and nuclear quadrupole splittings, as well as the electronic g tensors were evaluated for different model systems of the active site after complete geometry optimizations in the high-spin and broken-symmetry states. A comparison of the experimental data with the spectroscopic parameters computed for the models with unprotonated and protonated mu-oxo bridges shows best agreement between theory and experiment for a Mn2(mu-O)2(mu-OAc) core. The calculated Mn-Mn distances and 55Mn hfcs clearly support a dimanganese cluster with unprotonated mu-oxo bridges in the superoxidized state. Furthermore, it is shown that an interchange of the Mn(III) and Mn(IV) oxidation states in this trapped valence system leads to specific changes in the molecular and electronic structure of the manganese clusters. PMID- 15830217 TI - [An unusual foreign body in the upper gastrointestinal tract causing nonspecific abdominal pain]. AB - The risk of penetration of vena cava filters through the wall of the vena cava is estimated to be as high as 25%, although clinical symptoms are observed far less frequently in patients with this complication. Due to the close relationship between vena cava and duodenum, the latter can be injured by dislocated filters. We describe the presentation, evaluation, and treatment of a patient with a cava filter protruding into the duodenum, and we review the literature. PMID- 15830218 TI - The residue E351 is essential for the activity of human 21-hydroxylase: evidence from a naturally occurring novel point mutation compared with artificial mutants generated by single amino acid substitutions. AB - Congenital adrenal hyperplasia (CAH) [OMIM 201910] is a group of autosomal recessive disorders, caused in 90-95% of cases by a deficiency of steroid 21 hydroxylase due to mutations in the CYP21A2 gene. The functional and structural effects of a novel rare missense mutation (E351K) in CYP21A2 found in a male patient with simple virilizing CAH were studied. The novel E351K point mutation is located in the ERR triad of the 21-hydroxylase. The ERR triad is a glutamine arginine-arginine motif conserved in all cytochrome P450 sequences. The glutamate and first arginine residue are invariant in all P450 cytochrome enzymes, whereas the second arginine residue is present as arginine, histidine, or asparagine. Although the ERR triad is involved in some way to heme binding by the cytochrome P450 monooxygenases, the E351K mutation leads to severe but not complete loss of CYP21 enzyme activity. The functional analysis in COS-7 cells revealed a reduced conversion of 17-hydroxyprogesterone to 11-deoxycortisol of 1.1+/-0.5% (SD) and of progesterone to 11-deoxycorticosterone of 1.2+/-0.3% of wild-type activity. Analyzing the artificial mutants (E351D, E351I) of the E351 residue did not show a restoration of the in vitro 21-hydroxylase activity. These effects could be readily explained by structural changes induced by the mutations, which were rationalized by a three-dimensional-model structure of the CYP21 protein. The combination of in vitro enzyme function and computerized protein analysis of the E351 residue of the CYP21 protein provides experimental evidence for the ERR triad being a fundamental structural element of cytochrome P450 enzymes. PMID- 15830219 TI - Compensating for alcohol-induced impairment of control: effects on inhibition and activation of behavior. AB - RATIONALE: Studies have shown that expectations of alcohol-induced impairment can produce adaptive responses to alcohol that reduce the degree of behavioral impairment displayed. However, compensatory effects have been variable for cognitive and psychomotor tasks. OBJECTIVES: Using a model-driven approach to understanding behavioral control, the present study examined compensatory and impairment effects following the expectation and actual administration of alcohol on the ability to inhibit and activate behavior. METHODS: Social drinkers (N=17) performed a cued go/no-go task that required quick responses to go targets and inhibition of responses to no-go targets under four conditions: no treatment, alcohol expectancy (placebo), 0.45 g/kg alcohol, and 0.65 g/kg alcohol. RESULTS: Inhibitory and activational aspects of behavioral control were impaired by both active doses of alcohol. Alcohol slowed response activation and increased failures to inhibit responses in a dose-dependent manner. Subjects also displayed a small, but statistically significant, compensatory improvement in inhibitory control in response to the expectation of alcohol. Moreover, individual differences in the compensatory response were related to differences in alcohol impairment. Those who displayed greater compensatory responses to placebo displayed the least impairment in response to alcohol. CONCLUSIONS: By modeling behavioral control as the net effect of countervailing activational and inhibitory influences, the study suggests that fundamental mechanisms of control might not display uniform compensatory reactions to alcohol-related cues. Greater consideration of how alcohol-related expectancies shape behaviors under the drug should lead to a better understanding of individual differences with respect to risk for alcohol abuse. PMID- 15830220 TI - SSR181507, a dopamine D(2) receptor antagonist and 5-HT(1A) receptor agonist, alleviates disturbances of novelty discrimination in a social context in rats, a putative model of selective attention deficit. AB - RATIONALE: Selective attention deficit, characterised by the inability to differentiate relevant from irrelevant information, is considered to underlie many cognitive deficits of schizophrenia, and appears to be only marginally responsive to treatment with current antipsychotics. OBJECTIVES: We compared the activity of the putative atypical antipsychotic SSR181507 (a dopamine D(2) receptor antagonist and 5HT(1A) receptor agonist) with reference compounds, on disturbances of novelty discrimination in a social context in rats, a behavioural paradigm that putatively models selective attention deficit. METHODS: A first (familiar) juvenile rat was presented to an adult rat for a period (P1) of 30 min. A second (novel) juvenile was then introduced at the end of P1 for a period (P2) of 5 min. The ability of the adult rat to discriminate between the two juveniles, presented at the same time, was evaluated by measuring the ratio of the time spent in interaction with the novel vs the familiar juvenile during P2. RESULTS: Adult rats spent more time exploring the novel than the familiar juvenile. This novelty discrimination capacity was disrupted by: (1) parametric modification of the procedure (reduction of time spent in contact with the familiar juvenile during P1); (2) acute injection of psychotomimetics that are known to induce schizophrenia-like symptoms in humans, such as phencyclidine (PCP; 3 mg/kg, i.p.) and d-amphetamine (1 mg/kg, i.p.) and (3) neonatal treatment with PCP (three injections of 10 mg/kg, s.c.), a model based on the neurodevelopmental hypothesis of schizophrenia. The potential atypical antipsychotic SSR181507 (0.03-3 mg/kg, i.p.) and the atypical antipsychotics clozapine (0.1-1 mg/kg, i.p.) and amisulpride (1-3 mg/kg, i.p.) attenuated deficits in novelty discrimination produced by parametric manipulation and by acute or neonatal treatment with PCP. The typical antipsychotic haloperidol (up to 0.3 mg/kg, i.p.) attenuated only deficits in novelty discrimination produced by parametric modification. CONCLUSION: Collectively, these results suggest that SSR181507 can alleviate disturbances of novelty discrimination in a social context in rats, and that this paradigm may represent a suitable animal model of selective attention deficits observed in schizophrenia. PMID- 15830221 TI - Acute effects of alprazolam on risky decision making in humans. AB - RATIONALE: GABA-A receptor ligands, including benzodiapines, may induce disinhibitory effects that increase the probability of risky decision making. To date, few laboratory studies have examined the acute, dose-related effects of benzodiazepines on human risk-taking behavior. Recent data indicate that in the United States alprazolam is the benzodiazepine most frequently misused for recreational purposes. OBJECTIVES: The present study was designed to demonstrate a dose-response relationship between acute alprazolam administration and human risk taking. Furthermore, this investigation sought to examine: (1) the behavioral mechanisms that may be involved in changes in the probability of risky decision making related to alprazolam administration and (2) risk seeking-related personality variables that may predict drug effects on risk taking. METHODS: Using a laboratory measure of risk taking designed to address acute drug effects, 16 adults were administered placebo, 0.5, 1.0, and 2.0 mg alprazolam in a within subject repeated-measures design. The risk-taking task presented subjects with a choice between two response options operationally defined as risky and nonrisky. Data analyses examined subjective effects, response rates, distribution of choices between the risky and nonrisky option, trial-by-trial response probabilities, and personality correlates related to drug effects at the 2.0-mg dose. RESULTS: Alprazolam administration produced dose-related changes in subjective effects, response rates, and, most importantly, dose-dependently increased selection of the risky response option. The 2.0-mg dose increased the probability of making consecutive risky responses following a gain on the risky response option. Increases at 2.0 mg were related to a combination of personality scales that included high venturesomeness and novelty seeking and low harm avoidance. CONCLUSIONS: Alprazolam administration produced increases in human risk taking under laboratory conditions. In union with previous studies, the observed shift in trial-by-trial response probabilities suggests that sensitivity to consequences (e.g., oversensitivity to recent rewards) may be an important mechanism in the psychopharmacology of risky decision making. Additionally, risk seeking personality traits may be predictive of acute drug effects on risk-taking behavior. PMID- 15830223 TI - Effects of clonidine in the locus coeruleus on prefrontal- and hippocampal dependent measures of attention and memory in the rat. AB - RATIONALE: The locus coeruleus (LC) is the source of norepinephrine (NE) in the prefrontal cortex (PFC) and hippocampus and may influence cognitive functions of these areas. Chronic effects of LC-NE lesions do not correspond consistently with acute effects of systemic or intracortical injections of adrenergic agents. OBJECTIVE: These studies aim to manipulate LC activity pharmacologically and study acute effects on measures of attention and memory that depend on the PFC and hippocampus. METHODS: Rats were trained to criterion for one of three tasks: visuospatial reaction time (VSRT), a measure of attention sensitive to PFC lesions, delayed matching trained with retractable levers (DM-RL), and delayed nonmatching trained in radial mazes (DNM-RM), measures of spatial working memory sensitive to PFC and hippocampal lesions, respectively. LC activity was manipulated with bilateral 0.5-microl injections of the alpha-2 agonist clonidine (0, 1.1, 4.5, and 18 nmol). RESULTS: Clonidine produced significant dose dependent impairments of VSRT, affecting choice response time at the 18-nmol dose and choice accuracy at the 4.5- and 18-nmol doses. Clonidine had no effect on DMRL or DNM-RM at any of the doses tested. CONCLUSIONS: Reversible reduction of LC-NE activity by clonidine impaired measures of visuospatial attention sensitive to PFC lesions but were insufficient to affect PFC- or hippocampal-dependent measures of spatial working memory. These results are consistent with reports that LC-NE lesions produce chronic deficits in attention with little or no effect on measures of working memory. PMID- 15830222 TI - Plasma level-dependent effects of methylphenidate on task-related functional magnetic resonance imaging signal changes. AB - RATIONALE: Methylphenidate (MPH) is a dopamine and noradrenaline enhancing drug used to treat attentional deficits. Understanding of its cognition-enhancing effects and the neurobiological mechanisms involved, especially in elderly people, is currently incomplete. OBJECTIVES: The aim of this study was to investigate the relationship between MPH plasma levels and brain activation during visuospatial attention and movement preparation. METHODS: Twelve healthy elderly volunteers were scanned twice using functional magnetic resonance imaging (fMRI) after oral administration of MPH 20 mg or placebo in a within-subject design. The cognitive paradigm was a four-choice reaction time task presented at two levels of difficulty (with and without spatial cue). Plasma MPH levels were measured at six time points between 30 and 205 min after dosing. FMRI data were analysed using a linear model to estimate physiological response to the task and nonparametric permutation tests for inference. RESULTS: Lateral premotor and medial posterior parietal cortical activation was increased by MPH, on average, over both levels of task difficulty. There was considerable intersubject variability in the pharmacokinetics of MPH. Greater area under the plasma concentration-time curve was positively correlated with strength of activation in motor and premotor cortex, temporoparietal cortex and caudate nucleus during the difficult version of the task. CONCLUSION: This is the first pharmacokinetic/pharmacodynamic study to find an association between plasma levels of MPH and its modulatory effects on brain activation measured using fMRI. The results suggest that catecholaminergic mechanisms may be important in brain adaptivity to task difficulty and in task-specific recruitment of spatial attention systems. PMID- 15830224 TI - The dissociation of heroin-seeking patterns induced by contextual, discriminative, or discrete conditioned cues in a model of relapse to heroin in rats. AB - RATIONALE: The role of heroin-related stimuli in motivating the resumption of heroin use is not fully understood. OBJECTIVES: The objective was to characterize the relative importance of drug-related contextual stimuli, discriminative stimuli (DS), or discrete conditioned stimuli (CSs) on drug seeking when rats were reintroduced into the operant context after withdrawal. METHODS: Nose-poke responding by male rats was reinforced with intravenous heroin (0.05 mg/kg per infusion, 4-h session daily) under a progressive ratio schedule of reinforcement for 14 days. Each session began with the illumination of a green light in the active hole that served as DS. Each earned heroin injection was paired with a 5-s compound cue light and the sound of the infusion pump that served as the discrete CSs. RESULTS: Response rates of heroin seeking induced by the contextual stimuli were comparable to the average rates of responding during self-administration training, but rates induced by either DS or CSs were greater than those induced by the contextual stimuli alone (P<0.05). The responding induced by contingent presentations of CSs was higher than that of DS after extinction of instrumental behavior. The drug seeking induced by CSs can be maintained after 3 days extinction with DS in the original context, although the responding elicited by DS cannot be recovered after 3 days of extinction with CSs. CONCLUSIONS: The relapse to drug seeking can be elicited separately by environmental cues, heroin predictive DS, or discrete CSs in the same rat after withdrawal. PMID- 15830225 TI - Comparison of the effects of clozapine and 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) on progressive ratio schedule performance: evidence against the involvement of 5-HT1A receptors in the behavioural effects of clozapine. AB - RATIONALE: Performance on progressive ratio schedules has been proposed as a means of assessing the effects of drugs on the efficacy of reinforcers. A mathematical model (Killeen PR (1994) Mathematical principles of reinforcement. Behav Brain Sci 17:105-172) affords a basis for quantifying the effects of drugs on progressive ratio schedule performance. The model postulates a bitonic function relating response rate and ratio size. One parameter of the function, a, expresses the motivational effect of the reinforcer, whereas another parameter, delta, expresses the minimum time needed to execute a response, and is regarded as an index of 'motor capacity'. Previously we found that the atypical antipsychotic clozapine increased a, indicating an increase in reinforcer efficacy; a similar effect was observed with the 5-hydroxytryptamine (5-HT)(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). It has been suggested that some of clozapine's behavioural effects are mediated by agonistic action at 5-HT(1A) receptors. OBJECTIVE: This study was conducted to compare the effects of clozapine and 8-OH-DPAT on progressive ratio schedule performance. METHODS: Rats were trained under a time-constrained progressive ratio schedule (50-min sessions). In experiment 1, they received acute doses of clozapine (4 mg kg(-1)) and 8-OH-DPAT (100 microg kg(-1)), alone and in combination with the 5 HT(1A) receptor antagonist N-[2-(4-[2-methoxyphenyl]-1-piperazinyl)ethyl]-N-2 yridinylcyclohexanecarboxamide (WAY-100635; 30 microg kg(-1)). In experiment 2, the effects of clozapine (2, 4 and 8 mg kg(-1)) and 8-OH-DPAT (25, 50 and 100 microg kg(-1)) were compared between intact rats and rats whose 5-HTergic pathways had been ablated by 5,7-dihydroxytryptamine (5,7-DHT). RESULTS: In both experiments, clozapine and 8-OH-DPAT increased a and delta. In experiment 1, WAY 100635 abolished the effect of 8-OH-DPAT on a and delta, but did not alter clozapine's effects on these parameters. In experiment 2, the effects of clozapine and 8-OH-DPAT did not differ between sham-lesioned and 5,7-DHT-lesioned rats. CONCLUSIONS: The results confirm previous findings on the effects of clozapine and 8-OH-DPAT on progressive ratio schedule performance. 8-OH-DPAT's effects are probably mediated by post-synaptic 5-HT(1A) receptors; clozapine's effects are mediated by a different mechanism, which does not appear to involve 5 HT(1A) receptors and which does not depend upon an intact 5-HTergic pathway. PMID- 15830226 TI - Sex differences in neurocognitive functioning among abstinent recreational cocaine users. AB - RATIONALE: There are clear sex differences in the onset and course of cocaine addiction, females being at greater risk. Neurocognitive impairments associated with cocaine use are also well documented. However, there are no studies of sex differences in neurocognitive performance among cocaine users. OBJECTIVES: The present study was designed to examine, for the first time, putative sex-related variations in cognition among recreational cocaine users. It was predicted that female users would perform poorer compared with male users based on the worse effects of cocaine abuse in women. METHODS: A neuropsychological battery was administered to abstinent (72 h) recreational cocaine users (20 males and 20 females) and non-drug-using controls (19 males and 20 females) including tests of attention, executive functioning, verbal fluency (letter and category), verbal memory and visuospatial perception. RESULTS: Male users demonstrated poorer attention compared with male controls, and greater verbal recognition errors than male controls and female users. Female users did not differ significantly in any other cognitive domain compared with male users or female controls. These results remained after controlling for dose-dependent variables. General dose-dependent effects were found for attention, visuospatial perception and components of verbal memory. CONCLUSIONS: Whilst these data partially support the notion of sex related neurocognitive variation in short-term abstinence from recreational cocaine use, they do not suggest that sex is a major factor. PMID- 15830227 TI - Inhibition of calcium-independent phospholipase A2 activity in rat hippocampus impairs acquisition of short- and long-term memory. AB - RATIONALE: Phospholipase A(2) (PLA(2)) is a family of enzymes that cleave membrane phospholipids generating important lipid mediators in signal transduction. In rat hippocampal slices, both intracellular cytosolic Ca(2+) dependent PLA(2) (cPLA(2)) and Ca(2+)-independent PLA(2) (iPLA(2)) have been implicated in mechanisms of synaptic plasticity underlying memory processes. In mice, intraperitoneal injections of a selective iPLA(2) inhibitor impaired spatial learning. Accordingly, reduced cPLA(2) and iPLA(2) activities were found in postmortem hippocampus of patients with Alzheimer's disease. OBJECTIVE: This study investigates the effects of injections of PLA(2) inhibitors directly into rat hippocampus on the acquisition of short-term (STM) and long-term memory (LTM) of a one-trial step-down inhibitory avoidance (IA) task. METHODS: Wistar rats were bilaterally implanted with cannulae in the CA1 region of the dorsal hippocampus. After surgery, the rats received bilateral injections of a vehicle, or of dual cPLA(2) and iPLA(2) inhibitors (MAFP or PACOCF(3)), or a selective iPLA(2) inhibitor (bromoenol lactone) before training in IA. The animals were tested 1.5 h (for STM) and 24 h (for LTM) after training. RESULTS: Significant inhibition of iPLA(2) activity in rat hippocampus impaired acquisition of STM and LTM. Memory impairment did not result from neuronal death after iPLA(2) inhibition. Moreover, IA training per se increased significantly hippocampal PLA(2) activity. CONCLUSION: The present results suggest a functional effect of hippocampal PLA(2) on the neurochemistry of memory acquisition and support the hypothesis that reduced PLA(2) activity may contribute to memory impairment in Alzheimer's disease. PMID- 15830228 TI - Targeted invalidation of CCK2 receptor gene induces anxiolytic-like action in light-dark exploration, but not in fear conditioning test. AB - RATIONALE: Evidence suggests that gamma-aminobutyric acid (GABA) and cholecystokinin (CCK) have opposite roles in the regulation of anxiety. OBJECTIVES: The aim of our work was to study the behaviour of CCK(2) receptor deficient mice in light-dark exploration and fear conditioning tests. Moreover, the action of diazepam and methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3 carboxylate (DMCM), having the opposite effect on GABA(A) receptors, was evaluated on the exploratory behaviour in these mice. Expression levels of GABA(A) receptor subunit genes were also measured. METHODS: Light-dark exploration and fear conditioning tests were used to determine changes in anxiety of mice. The action of diazepam (0.5-2 mg/kg i.p.) and DMCM (0.25-1 mg/kg i.p.) was studied in the light-dark box. The effect of DMCM was also evaluated in the motor activity test to demonstrate that its anti-exploratory action was not related to motor suppression. Expression levels of GABA(A) receptor subunit genes were determined by means of real-time polymerase chain reaction (qRT-PCR). RESULTS: Female mice lacking CCK(2) receptors displayed increased exploratory activity in the light-dark box compared to their wild-type (+/+) littermates. Locomotor activity in the motility boxes and the intensity of freezing did not differ in wild-type (+/+) and homozygous (-/-) mice. Treatment with diazepam (0.5 mg/kg) increased the number of transitions in wild-type (+/+) animals, whereas in homozygous (-/-) mice diazepam (0.5-2 mg/kg) reduced exploratory activity. Administration of DMCM (0.25-1 mg/kg) induced an anxiogenic-like effect in homozygous (-/-) mice, but did not change their locomotor activity. Gene expression analysis established a 1.6-fold increase in the expression of the alpha2 subunit of GABA(A) receptors in the frontal cortex of homozygous (-/-) mice. CONCLUSION: Genetic invalidation of CCK(2) receptors induced an anxiolytic like action in exploratory, but not in conditioned models of anxiety. The observed reduction in anxiety in homozygous (-/-) mice is probably related to an increased function of GABAergic system in the brain. PMID- 15830229 TI - Decreased striatal dopamine transporter binding assessed with [123I] FP-CIT in first-episode schizophrenic patients with and without short-term antipsychotic induced parkinsonism. AB - RATIONALE: Drug-induced parkinsonism (DIP) is one of the main causes of treatment drop-out in schizophrenic patients causing a high incidence of relapse that leads patients to a bad clinical prognosis. The dopaminergic nigrostriatal pathway is involved in the movement control, so the study of the dopamine transporter (DAT) could be of great value to determine its implication in the appearance of DIP. OBJECTIVE: The goal of the study is to determine the striatal DAT binding assessed with [(123)I] FP-CIT SPECT in first-episode neuroleptic-naive schizophrenic in-patients with DIP after short-term antipsychotic treatment. METHOD: The [(123)I] FP-CIT binding ratios of ten schizophrenic in-patients who developed DIP during the first 4-week period of risperidone treatment (6+/-2 mg/day) were compared with ten schizophrenic in-patients treated with the same doses of risperidone and who do not developed DIP and with ten age-matched healthy subjects. Quantitative analyses of SPECTs were performed using regions of interest located in caudate, putamen and occipital cortex. Parkinsonism was assessed by the Simpson-Angus Scale and the psychopathological status by the Clinical General Impression and Positive and Negative Syndrome Scales. RESULTS: Whole striatal [(123)I] FP-CIT binding ratios were significantly lower in patients with and without DIP than in healthy subjects (p<0.001). This was also observed in whole putamen (p<0.001) and caudate nucleus (p<0.001). Females showed higher whole striatal [(123)I] FP-CIT binding ratios than males (p<0.05). No differences in psychopathological scales were observed between patients with and without DIP. CONCLUSION: Our first-episode schizophrenic patients with and without DIP after short-term risperidone treatment have a decreased striatal DAT binding assessed with [(123)I] FP-CIT. This alteration could be related to the schizophrenic disease or may be secondary to the antipsychotic treatment. PMID- 15830230 TI - Effector antagonism by the regulators of G protein signalling (RGS) proteins causes desensitization of mu-opioid receptors in the CNS. AB - RATIONALE: In cell culture systems, agonists can promote the phosphorylation and internalization of receptors coupled to G proteins (GPCR), leading to their desensitization. However, in the CNS opioid agonists promote a profound desensitization of their analgesic effects without diminishing the presence of their receptors in the neuronal membrane. Recent studies have indicated that CNS proteins of the RGS family, specific regulators of G protein signalling, may be involved in mu-opioid receptor desensitization in vivo. OBJECTIVE: In this work we review the role played by RGS proteins in the intensity and duration of the effects of mu-opioid receptor agonists, and how they influence the delayed tolerance that develops in response to specific doses of opioids. RESULTS: RGS proteins are GTPase-activating proteins (GAP) that accelerate the hydrolysis of GalphaGTP to terminate signalling at effectors. The GAP activity of RGS-R4 and RGS-Rz proteins restricts the amplitude of opioid analgesia, and the efficient deactivation of GalphazGTP subunits by RGS-Rz proteins prevents mu receptor desensitization. However, RGS-R7 proteins antagonize effectors by binding to and sequestering mu receptor-activated Galphai/o/z subunits. Thus, they reduce the pool of receptor-regulated G proteins and hence, the effects of agonists. The delayed tolerance observed following morphine administration correlates with the transfer of Galpha subunits from mu receptors to RGS-R7 proteins and the subsequent stabilization of this association. CONCLUSION: In the CNS, the RGS proteins control the activity of mu opioid receptors through GAP-dependent (RGS R4 and RGS-Rz) as well as by GAP-independent mechanisms (RGS-R7). As a result, they can both antagonize effectors and desensitize receptors under certain circumstances. PMID- 15830231 TI - Peripheral-type benzodiazepine receptor binding sites in platelets of patients with panic disorder associated to separation anxiety symptoms. AB - RATIONALE: Although it is still a matter of debate whether panic disorder (PD) and separation anxiety (SA) are associated or causally linked disorders, some investigators have suggested that SA may be a specific subtype of panic agoraphobic spectrum. Several psychiatric disorders, including PD, are associated with lower levels of peripheral-type benzodiazepine receptor (PBR). OBJECTIVES: The aim of the present study was to evaluate the kinetic binding parameters of the specific PBR ligand, PK 11195, in platelets from patients with PD in relation to the presence and severity of adulthood SA. METHODS: Using the specific radioligand, [(3)H] PK 11195, the kinetic binding parameters of PBR were determined on platelet membranes of 27 adult outpatients with a DSM-IV diagnosis of PD and 18 healthy controls. Patients were assessed with the SCID-I, the Panic Disorder Severity Scale, the Structured Clinical Interview for Separation Anxiety Symptoms and the Adult Separation Anxiety Checklist. RESULTS: PD patients had significantly lower PBR density than controls. However, the lower density was only evident in the subgroup of PD patients who also fulfilled the DSM-IV criteria for adult separation anxiety disorder. PBR density was negatively correlated with each of the two SA scales total scores. CONCLUSIONS: Patients with SA symptoms had significantly lower densities of PBRs. PBR expression might become a useful biological marker of these two associated conditions. PMID- 15830232 TI - Influence of cocaine self-administration on learning related to prefrontal cortex or hippocampus functioning in rats. AB - RATIONALE: Individuals who abuse cocaine have cognitive deficits, particularly in functions associated with the orbitofrontal cortex. It is not clear to what extent the impact of cocaine on cognitive functioning is related to its role as a behavioral reinforcer. A preclinical means to investigate this issue is to use a yoked-triad procedure in which sets of three animals either contingently self administer cocaine or receive passive administration of cocaine or saline in a noncontingent manner. OBJECTIVE: Using this procedure, we assessed cocaine's effect on learning that requires a functionally intact prefrontal cortex (prelimbic or insular/orbital subregions) or hippocampus. METHODS: Rats self administering 1-mg/kg unit doses of cocaine responded under a fixed-ratio 5, time out 20-s schedule of drug delivery. Testing took place in a radial-arm maze within the first 30 min after 2-hr drug sessions ended, beginning after 2.5 months of cocaine or saline exposure. RESULTS: Rats self-administering cocaine earned 14-18 infusions on average throughout different phases of the study. In groupwise comparisons, learning in the visually guided delayed win-shift (prelimbic prefrontal cortex-related) and win-shift (hippocampus-related) tasks was not influenced by contingent or noncontingent cocaine exposure. Session latency, though, was shorter in both cocaine-exposed groups during the win-shift task. During the odor-guided delayed win-shift task (insular/orbital prefrontal cortex-related), learning was disrupted in rats self-administering cocaine, with no influence of noncontingent cocaine exposure. CONCLUSIONS: Based on these and previous findings, learning related to functioning of the insular/orbital prefrontal cortex and amygdala is the most consistently disrupted in cocaine intoxicated rats after long-term drug exposure. PMID- 15830233 TI - Reinforcing effects of oral Delta9-THC in male marijuana smokers in a laboratory choice procedure. AB - RATIONALE: Oral Delta-9-tetrahydrocannabinol (Delta(9)-THC; Marinol) is medically available for the treatment of nausea associated with cancer chemotherapy and for wasting syndromes related to HIV/AIDS. Little is known about its reinforcing effects. OBJECTIVE: This study was conducted to characterize the reinforcing effects of oral Delta(9)-THC in experienced marijuana smokers under controlled laboratory conditions. METHODS: Ten healthy male marijuana users completed this 17-day residential study. On days 2, 6, 10, and 14, at 0900 h, participants received a "sample" oral dose of Delta(9)-THC (0, 10, 20 mg) and an alternative reinforcer, a 2 US dollars voucher (redeemable for cash at study's end). Over the next 3 days, they had 11 opportunities to self-administer either the sampled dose of Delta(9)-THC or to receive a 2 US dollars voucher. RESULTS: Participants chose active Delta(9)-THC (10 and 20 mg) more often than placebo ( or = 220 mg/dl or TG > or = 150 mg/dl. The cumulative incidences of ESRD, per 1000 screenees, were 1.12 for those without dyslipidemia and 2.53 for those with dyslipidemia. The adjusted hazard ratio (95% confidence interval) for dyslipidemia was 0.856 (0.484-1.516) for men and 1.260 (0.661-2.400) for women; neither was significant when adjustment was made for age, systolic blood pressure, diastolic blood pressure, body mass index, creatinine clearance, diabetes mellitus, and proteinuria. CONCLUSIONS: The present study showed dyslipidemia to be an insignificant predictor of development of ESRD in the general Okinawa population. PMID- 15830274 TI - Correction of CAPD catheter displacement using alpha-replacement method. AB - BACKGROUND: Malposition of the tip of a continuous ambulatory peritoneal dialysis (CAPD) catheter may impede a dialysis, and an invasive method, such as open or laparoscopic surgery, is necessary to correct its position. The alpha-replacement method is a nonsurgical procedure; however, the reported clinical experience has been limited. We thus reviewed our 19 cases to clarify the efficacy of this method. METHOD: The alpha-replacement method was used in 19 patients [mean age, 50 +/- 14 years old; male/female ratio, 13/6; duration of catheter insertion, 23.7 +/- 23.9 months; total, 29 times in 19 patients (once in 13 patients, twice in 3, three times in 2, four times in 1 patient)]. All patients received a swan neck J-type catheter. The procedure was carried out by using a special guidewire within 1-2 weeks after dislocation of the tips. RESULTS: The procedure was successfully performed 22 times in 14 patients (success rate, 78% in each performance). Required time was only 5-10 min in most successful cases. No evident complications were encountered in any case. CONCLUSIONS: The alpha replacement method for a displaced CAPD catheter proved to be effective and safe. PMID- 15830275 TI - Identification of a novel insertion mutation in GATA3 with HDR syndrome. AB - Recently, a member of the GATA-binding family of transcription factors was shown to be involved in human hypoparathyroidism, sensorineural deafness, and renal abnormality (HDR) syndrome. We report here a Japanese family in which two of the members are affected with HDR syndrome. Sequence analysis of GATA3 showed a heterozygous novel mutation in this family: an unusual mutation at exon 3 (709insC) resulting in a premature stop at codon 302 with a loss of both of the zinc finger domains. PMID- 15830276 TI - Steroid resistance in prolonged type I membranoproliferative glomerulonephritis and accelerated disease remission after steroid withdrawal. AB - Two cases of severe proteinuria and hypocomplementemia were referred to our out patient clinic for continuous follow-up. Initial onset of clinical symptoms was at the age of 15 years in both cases. They had already been diagnosed as type I membranoproliferative glomerulonephritis (type I MPGN) by renal biopsy when oral prednisolone administration had been initiated. Several courses of steroid pulse therapy were performed for the flares of the disease, resulting in only temporary amelioration of renal symptoms. Percutaneous renal biopsy was performed during admission on both cases, showing severe glomerular and tubulointerstitial damages, in addition to typical type I MPGN findings such as mesangial cells and matrix interposition and subendothelial deposits. Because the continuous administration of steroid for more than 10 years did not ameliorate the clinical symptoms, steroid was markedly reduced or stopped in these cases. Such withdrawal from steroid therapy accelerated the amelioration of renal symptoms, including decrease in proteinuria, elevation of plasma protein and complement levels, and disappearance of generalized edema. The clinical courses of these cases indicate clinical choice of withdrawal from steroid therapy as one of the treatments in prolonged type I MPGN, which presents in childhood and shows steroid resistance. PMID- 15830277 TI - The increase of antiglomerular basement membrane antibody following pauci-immune type crescentic glomerulonephritis. AB - A 50-year-old woman was admitted because of high fever and fatigue. Proteinuria, hematuria, and elevated BUN (47.8 mg/dl) and creatinine (3.4 mg/dl) suggested rapidly progressive glomerulonephritis. The serological study revealed all negative results for rheumatoid factor, antinuclear antibody, serum cryoglobulins, MPO-ANCA, PR3-ANCA, and anti-streptolysin O. Antiglomerular basement membrane (GBM) antibody, as assessed by ELISA, was 11 EU (normal, <10). Kidney biopsy on the eighth hospital day demonstrated pauci-immune-type crescentic glomerulonephritis without ANCA. Methylprednisolone pulse therapy (500 mg/day, 3 days) and 45 mg/day prednisolone orally were started. At 3 weeks after kidney biopsy, the anti-GBM antibody value increased from 11 EU/ml to 116 EU/ml, and MPO and PR3-ANCA were still negative. HLA type was DR8 and DR 15(2), with a genotype of HLA-DRB1*08021 and HLA-DRB1*15011. The present case suggests that HLA DR15 plays an important role on antibody production against alpha 3(IV) NC1 autoantigen after severe nephritis or tissue damage. PMID- 15830278 TI - A case of necrotizing glomerulonephritis presenting with nephrotic syndrome associated with pulmonary cryptococcosis. AB - We describe a 68-year-old man with necrotizing glomerulonephritis who presented with nephrotic syndrome accompanied by pulmonary cryptococcosis. He developed rheumatoid arthritis in July 1999 and was treated with low-dose prednisolone. He was admitted to our hospital on November 22 following the appearance of bilateral leg edema in October 2000. Laboratory tests at presentation revealed nephrotic syndrome with renal impairment. Renal biopsy specimens revealed necrotizing glomerulonephritis with crescent, but immunofluorescence study showed lack of staining for immunoglobulins or complement components. Chest X-ray and CT showed abnormal shadows in the right upper lung field, and Cryptococcus neoformans was isolated in a transbronchial lung biopsy. After the diagnosis of pulmonary cryptococcosis was made, the patient was treated with 200 mg/day fluconazole. The pulmonary abnormal shadows immediately improved and urinary protein excretion dramatically decreased. A second renal biopsy, performed about 2 months after the first biopsy, showed disappearance of crescent. Electron microscopic examination of the second renal biopsy showed partial effacement of foot processes without electron-dense deposits. Our findings suggest that necrotizing glomerulonephritis with nephrotic syndrome in this patient represented pauci-immune T-cell-mediated injury related to pulmonary cryptococcosis. PMID- 15830279 TI - Predominant tubulointerstitial nephritis in a patient with systemic lupus nephritis. AB - In most cases of systemic lupus erythematosus (SLE), glomerular lesions are the main renal complication. Although tubulointerstitial lesions are often associated with severe glomerular lesions, predominant or isolated tubulointerstitial injury in the presence of minimal glomerular abnormalities with SLE, so-called predominant tubulointerstitial lupus nephritis, is rare. Only ten cases are reported in the English literature. Herein, we describe the case of a 64-year-old man with SLE who presented with acute renal deterioration attributable to acute tubulointerstitial nephritis. Renal biopsy showed diffuse infiltration of inflammatory mononuclear cells in the interstitium and tubulitis without significant glomerular lesions. Immunofluorescence study revealed positive staining for IgG, C3, and C1q along the renal tubular basement membrane (TBM). Electron microscopy also showed electron-dense deposits in the TBM. Other causes of tubulointerstitial injury, such as drug use and infection, were ruled out. Taking these findings together with the presence of antitubular basement membrane antibody, predominant tubulointerstitial lupus nephritis was diagnosed. Treatment with oral corticosteroids for 6 weeks improved renal function. Even after tapering of the corticosteroid, renal function and serological markers of SLE activity have remained stable in this patient for more than 12 months. PMID- 15830280 TI - A case of encapsulating peritoneal sclerosis at the clinical early stage with high concentration of matrix metalloproteinase-2 in peritoneal effluent. AB - In encapsulating peritoneal sclerosis (EPS), matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases are involved in the remodeling of peritoneal tissue. We measured the MMP-2 concentration in the peritoneal effluents of a patient with EPS who discontinued continuous ambulatory peritoneal dialysis (CAPD) therapy because of ultrafiltration failure and/or underdialysis. First, we report a 58-year-old female patient who discontinued CAPD therapy because of underdialysis. Several months after cessation of CAPD, she complained of slightly blood-colored ascites and had an elevated level of C-reactive protein (CRP) in plasma. She was diagnosed as having clinical early-stage EPS. Peritoneal effluents drained from this case, and from 11 patients who discontinued CAPD therapy because of ultrafiltration failure and/or underdialysis, and who underwent peritoneal lavage with 1.5% dextrose peritoneal dialysis fluid for several months, were analyzed by gelatin zymography. MMP-2 concentration was also measured by enzyme-linked immunosorbent assay (ELISA). MMP-2 concentration in peritoneal effluent of this patient was highest compared with that of the other patients. There was some tendency of a positive correlation between MMP-2 concentration per 1 g protein and D/Pcr, and was negative correlation between MMP 2 concentration per 1 g of protein and D/D0 glucose. We concluded that MMP-2 is involved in the peritoneal remodeling of long-term CAPD therapy and the progression of EPS. PMID- 15830281 TI - Updated treatment approach to hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide, and its incidence will further increase, to reach a plateau in 2015-2020. The natural history of the disease is quite well known, except for its early stages, because the majority of patients at this stage are treated with radical approaches. Staging systems are key to predict the prognostics of patients with cancer, to stratify the patients according to prognostic variables in the setting of clinical trials, and to guide the therapeutic approach. The current knowledge of the disease, however, is not sufficient for recommending a staging system to be used worldwide. The conventional staging systems-Okuda stage, and TNM stage-have shown important limitations for classifying patients. Several new systems have been recently proposed, but only three of them have been validated. The Barcelona Clinic Liver Cancer (BCLC) staging classification links the stage of the disease to a specific treatment strategy. The Japan Integrated Staging (JIS) score has been proposed and used in Japan, although it needs Western validation. The Cancer of the Liver Italian Program (CLIP) score is mainly proposed for patients with advanced tumors. Early detection of HCC through surveillance programs allows the application of potentially curative therapies, such as resection, liver transplantation, and percutaneous ablation in patients with early tumors. The applicability of these treatments varies according to geographical distribution: from 50% to 70% of cases in Japan; 25% to 40% of cases in Europe and the United States; and fewer than 10% in Africa. There are no randomized controlled trials (RCTs) comparing any of the three major therapies. These studies are not feasible in the West. Therefore, there is no firm evidence to establish the optimal first line treatment for small single HCC in patients with well-preserved liver function. Resection and transplantation achieve the best outcomes in well selected candidates (5-year survival of 60%-70%), and compete as the first option from an intention-to-treat perspective. If surgery is precluded, local, nonsurgical therapies are applied. Percutaneous treatments provide good results (5-year survival of 40%-50%), but are unable to achieve response rates and outcomes comparable to those for surgical treatments, even when applied as the first option. Radiofrequency thermal ablation provides slightly better objective response rates than ethanol injection, but no survival advantages have been fully demonstrated. The remaining treatments have been assessed in the setting of around 70 RCTs conducted during the past 25 years. Chemoembolization has been shown to provide modest survival advantages in two RCTs and a metaanalysis, and is currently the mainstay of treatment in 10% of the whole HCC population. The ideal candidates for this option are patients with well-preserved liver function (Child-Pugh class A) and multinodular asymptomatic tumors without vascular invasion. Further RCTs are needed to assess the best chemotherapeutic agent and the ideal re-treatment schedule. There is no firstline option for patients with advanced HCC (vascular invasion, extrahepatic spread, or cancer-related symptoms). Systemic doxorubicin provides partial responses in 10% of cases, without proven survival advantages, and well-known treatment-related complications. Several other treatments, such as immunotherapy, internal radiation, tamoxifen, or anti-androgen agents, have not shown any relevant anti tumoral effect or survival benefit. New drugs, such as tyrosine kinase inhibitors and anti-angiogenic agents, are currently being tested in the setting of clinical trials. PMID- 15830282 TI - Gastric epithelial cell turnover and mucosal protection in Japanese children with Helicobacter pylori infection. AB - BACKGROUND: In adults, epithelial cell proliferation and apoptosis of the gastric mucosa are induced by Helicobacter pylori infection and are associated with gastric atrophy or gastric carcinoma. In children, there are few studies about such epithelial changes. To elucidate the role of H. pylori infection in gastric mucosal inflammation, we immunohistochemically examined gastric mucosa of Japanese children. METHODS: Biopsy specimens obtained from the gastric antrum and corpus of H. pylori-infected (n = 13) and noninfected children (n = 15) were studied for immunolocalization of Ki-67, single-strand DNA, manganese superoxide dismutase (Mn-SOD), and CD68, and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling. In 10 patients with successful eradication, pre- and posttreatment results were compared. RESULTS: In both gastric antrum and corpus, neutrophil and mononuclear cell infiltration, epithelial cell proliferation, and apoptosis significantly increased in H. pylori-infected patients, predominantly in the antrum. In the antrum of H. pylori-infected patients, there was positive correlation between the degrees of neutrophil infiltration and cell proliferation (P < 0.05) or apoptosis (P < 0.05). H. pylori eradication improved mucosal inflammation, cell proliferation (P < 0.001), and apoptosis (P < 0.01) in the antrum. Mn-SOD immunoreactivity and CD68-positive macrophages in the antrum, which significantly increased in H. pylori-infected patients, decreased after the eradication. CONCLUSIONS: H. pylori infection induced gastric mucosal inflammation and epithelial cell turnover in children. Moreover, gastric mucosal defense mechanism against H. pylori infection was activated. H. pylori eradication in childhood might prevent the accumulation of gastric epithelial cell damage. PMID- 15830283 TI - Estimation of the malignant potential of gastrointestinal stromal tumors: the value of contrast-enhanced coded phase-inversion harmonics US. AB - BACKGROUND: Recently, contrast agents for ultrasonography (US) such as Levovist have been introduced for routine clinical use. The contrast-enhanced US with Levovist permits evaluation of the intratumoral vascularity of hepatic and pancreatic tumors and is useful for their differential diagnosis. The purpose of the present study was to assess tumor vessels and the parenchymal flow of gastrointestinal stromal tumors (GISTs) by contrast-enhanced coded phase inversion harmonic US and to evaluate whether vascularity is related to the malignant grade of the GISTs. METHODS: Thirteen patients with GISTs were included in the present study. Tumors were observed in a real-time fashion of contrast enhanced coded phase-inversion harmonic US after the injection of Levovist (400 mg/ml). The vascular patterns were compared with tumor size, histological diagnosis, KIT mutations, and clinical findings such as metastasis. RESULTS: The contrast-enhanced US images of the GISTs were classified into two types according to the blood flow area of the tumors as seen by real-time continuous imaging of the tumor vessels. The image pattern "Poor" represented vessels flowing only in the peripheral part of the tumor, and "Rich" represented abundant vessels flowing from the periphery to the central part of the tumor. According to the contrast enhanced US images, five GISTs were classified as "Poor" and the others as "Rich." Based on the final diagnosis, all tumors with "Poor" images were determined to be benign GISTs, and the rest tumors except one with "Rich" images were determined to be malignant GISTs. CONCLUSIONS: Contrast-enhanced US image is more closely correlated with the final diagnosis than the histological findings. PMID- 15830284 TI - The role of video capsule endoscopy for evaluating obscure gastrointestinal bleeding: usefulness of early use. AB - BACKGROUND: We report our preliminary experience with the use of video capsule endoscopy (VCE) in 64 patients with obscure gastrointestinal bleeding (OGIB) and suspected small intestine disease. METHODS: To be eligible for VCE, patients had to have undergone upper endoscopy, small bowel series, and colonscopy without discovering any source of bleeding. To find the best timing to perform VCE, the patients were retrospectively divided in two groups of 32 cases each: group 1 with patients who had been submitted to VCE within 15 days from OGIB diagnosis, and group 2 with patients who had been submitted to VCE at least 15 days after OGIB diagnosis. RESULTS: Lesions were found by VCE in 29 (91%) in group 1: angioectasia-like lesions of the small bowel in 12, some erosions of the ileum without signs of bleeding in 14, a polyp with erosions in 1, and a bleeding site where the surgery showed a tumor of the ileum in 2 patients. In 2 cases, VCE missed showing two small tumors that were revealed by laparoscopy in 1 case and by push enteroscopy in the other. In group 2, lesions were found by VCE in 11 (34%): angioectasia-like lesions of the small bowel in 6, some erosions in 3, a short segmental stenosis in 1, and two polyps in 1. In 1 case, VCE missed showing a small polyp in the jejunum that was revealed by push enteroscopy. In none of these cases was a bleeding site identified. VCE was well tolerated and able to acquire good images in patients with OGIB. It showed lesions in 91% of the patients in group 1 and 34% of cases in group 2. CONCLUSIONS: Our data suggest that the optimal timing to perform VCE is within a few days after the occurrence of bleeding, possibly within 2 weeks. PMID- 15830285 TI - Gabexate mesilate, a synthetic protease inhibitor, attenuates carbon tetrachloride-induced liver injury in rats. AB - BACKGROUND: Gabexate mesilate, a synthetic protease inhibitor, is used to treat acute pancreatitis and disseminated intravascular coagulation because it inhibits various serine proteases; however, whether gabexate mesilate prevents acute liver failure has not yet been studied. The aim of the present study was to investigate the effect of gabexate mesilate in carbon tetrachloride (CCl4)-induced liver injury in rats. METHODS: Acute hepatic failure was induced by administration of CCl4 intragastrically to male Sprague-Dawley rats. The effects of gabexate mesilate were examined in terms of serum transaminase levels, liver histology, and the prognosis of rats. RESULTS: Gabexate mesilate treatment significantly decreased the elevation of serum transaminase levels and improved liver histology 24 h after the administration of CCl4 (0.2 ml/100 g rat weight). Plasma tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) decreased significantly in the gabexate mesilate-treated rats compared with saline-treated rats. Gabexate mesilate treatment also significantly improved survival rate after a lethal dose of CCl4 (0.5 ml/100 g rat weight) from 0% to 20%. CONCLUSIONS: Gabexate mesilate treatment attenuated CCl4-induced liver injury via a suppression of proinflammatory cytokine production. In addition, these investigations suggest that gabexate mesilate treatment may provide therapeutic strategies for human acute liver failure. PMID- 15830286 TI - Analysis of factors influencing hepatocellular carcinoma detection: efficient use of computed tomography during arterial portography and during hepatic arteriography. AB - BACKGROUND: This study was performed to investigate the situations in which computed tomography (CT) combined with arterial portography and hepatic arteriography surpassed dynamic CT in the detection of hepatocellular carcinoma. METHODS: Computed tomography combined with arterial portography and hepatic arteriography was performed on 137 patients with chronic hepatitis (92 men and 45 women; mean age, 66.5 years) with hepatocellular carcinoma (HCC) as revealed or suspected by dynamic CT. We analyzed the clinical factors leading to the discovery of additional HCC lesions on CT combined with arterial portography and hepatic arteriography that were undetected by dynamic CT. RESULTS: Computed tomography combined with arterial portography and hepatic arteriography detected additional HCC lesions that had not been revealed by dynamic CT in 33 of 137 patients. Univariate analysis revealed that in the event of HCC recurrence (vs. primary), multiple HCC lesions detected by dynamic CT (vs. single) and decreased liver function (Child's classification B/C vs. A) significantly favored the additional detection of HCC lesions. Multivariate logistic regression indicated that recurrence was the strongest predicting factor for finding additional lesions on computed tomography combined with arterial portography and hepatic arteriography. CONCLUSIONS: Computed tomography combined with arterial portography and hepatic arteriography is capable of finding additional HCC lesions undetectable by dynamic CT, especially in advanced cases such as HCC recurrence, which may affect the choice of treatment. PMID- 15830287 TI - Clinicopathological features of hepatocellular carcinomas (HCCs) arising in patients without chronic viral infection or alcohol abuse: a retrospective study of patients undergoing hepatic resection. AB - BACKGROUND: This study was carried out to clarify the etiology and clinicopathological features of hepatocellular carcinomas (HCCs) arising in patients without chronic viral infection or alcohol abuse. METHODS: HCC patients who underwent resection were divided into three groups: a non-B non-C (NBNC) group (n = 13), who were seronegative for hepatitis B surface antigen (HBs Ag) and anti-hepatitis C antibody (HCV Ab), excluding a history of alcohol abuse; a B group (n = 25), who were seropositive for HBs Ag only; and a C group (n = 116), who were seropositive for HCV Ab only. We analyzed the features of tumor- and host-related factors and the outcome of the NBNC group. RESULTS: Hepatic inflammation and fibrosis were less severe in the NBNC group than in the other groups. There were no significant differences in tumor-related factors, except for higher serum levels of alpha-fetoprotein in the NBNC group. Recurrence rates and disease-free survivals were comparable among the three groups. The NBNC group comprised a greater population with one or two recurrent hepatic lesions (P < 0.05), and indocyanine green retention rates and fibrosis scores were preserved after the initial hepatectomy. The NBNC group had higher resection rates for intrahepatic recurrences (75.0%) than the other groups (21.1% and 22.2% in groups B and C, respectively; P < 0.05 and P < 0.05). The survival rate after the initial hepatectomy or detection of the recurrent lesions was significantly better in the NBNC group (both 100% at 5 years) than those in groups B and C (P < 0.05). CONCLUSIONS: NBNC patients maintained good liver function following the initial hepatectomy, and tended to have one or two recurrent lesions. These biological advantages provided NBNC patients more opportunities for repeat resection of intrahepatic recurrences, which may lead to a favorable outcome. PMID- 15830288 TI - Monitoring of hepatocellular carcinoma, following proton radiotherapy, with contrast-enhanced color Doppler ultrasonography. AB - BACKGROUND: We have reported that proton radiotherapy for hepatocellular carcinoma (HCC) is a safe and effective therapeutic option. However, it is difficult to evaluate its effect in certain cases. Recently, it has been reported that the usage of contrast-enhanced color Doppler ultrasonography (CECDU) can improve diagnostic accuracy, both in terms of the presence of hepatic tumor and in the evaluation of treatment. The aim of this study was to determine the usefulness of CECDU in assessing the therapeutic response of HCC treated with proton radiotherapy. METHODS: Twenty-two patients treated with the proton radiotherapy were studied. We inspected HCC lesions by CECDU, before and after the irradiation, over time. The magnitude of blood flow in the HCC was quantified on still images by CECDU. The ratio of the number of color pixels against that of the total number of pixels in the tumor area was defined as the tumor blood flow ratio (TBFR). RESULTS: Immediately after the proton treatment, a transient increase of blood flow in the tumor was recognized in more than half of the patients, while the TBFR was unchanged or decreased in the remaining patients. At longer periods after irradiation, the TBFR in all HCCs gradually decreased, and this reduction of TBFR was statistically significant from 9 months after irradiation. These findings are consistent with those obtained previously by computed tomography (CT) as well as magnetic resonance imaging (MRI). CONCLUSIONS: We propose CECDU as a useful diagnostic option for the evaluation of HCC treated with proton radiotherapy. PMID- 15830289 TI - Estimation of the stent placement above the intact sphincter of Oddi against malignant bile duct obstruction. AB - BACKGROUND: In endoscopic biliary stenting against malignant biliary obstruction, stent blockage remains as an important problem. Stent blockage occurs as a result of bacterial adherence to the inner wall of the stent. We evaluated the stent placement above the intact sphincter of Oddi to retain the function of the sphincter of Oddi as a bacteriological barrier. METHODS: Sixteen patients with malignant biliary obstruction were assessed as the patients with the stent above the intact sphincter of Oddi. Sixteen patients with malignant biliary obstruction were assessed as the patients with the conventional stent placement across the sphincter of Oddi. Tannenbaum 10-Fr. stents were used in both the groups. RESULTS: The median patency periods of the stent were 255 days (25th to 75th percentiles, 212-454 days; range, 39-454 days) for the group of the stents placed above the sphincter of Oddi and 82 days (25th to 75th percentiles, 48-131 days; range, 22-196 days) for the group of the stents placed across the sphincter of Oddi, respectively, with significant difference (P = 0.0001). The occlusion rates of stents placed above and across the sphincter of Oddi were 37.5% and 93.8%, respectively, with significant difference (P = 0.0008). The dislocation rates of the stent were 0% and 6.3%, respectively (not significant). CONCLUSIONS: Placement of the stent above the intact sphincter of Oddi was associated with longer stent patency and lower occlusion rate. PMID- 15830290 TI - Nutritional factors and risk of pancreatic cancer: a population-based case control study based on direct interview in Japan. AB - BACKGROUND: Few epidemiologic studies have examined the role of nutrient intake in the development of pancreatic cancer in Japan. We addressed this association in a population-based case-control study. METHODS: The cases were 109 patients who were newly diagnosed with pancreatic cancer between January 2000 and March 2002, and controls were selected by a random procedure from the general population. Data on dietary intake were collected by in-person interview, with the use of a food-frequency questionnaire. The risk of pancreatic cancer associated with nutrient intake was estimated by using the odds ratios (ORs) and 95% confidence intervals (CIs) derived from a conditional logistic model. RESULTS: A statistically positive trend in risk was observed with increasing cholesterol intake, with subjects in the highest tertile experiencing a two fold increased risk (OR, 2.06; 95% CI, 1.11-3.85; Ptrend = 0.02). Vitamin C intake was negatively associated with risk of pancreatic cancer. The OR was 0.45 (95% CI, 0.22-0.94) for subjects in the highest tertile compared to the lowest tertile (Ptrend = 0.04). CONCLUSIONS: Our study indicates that high cholesterol intake is significantly associated with an increased risk of pancreatic cancer and that high vitamin C intake decreases the risk of pancreatic cancer. PMID- 15830291 TI - Post-transplant IVC occlusion and thrombosis treated with tPA, heparin, and sharp recanalization. AB - Complete inferior vena cava (IVC) thrombosis can be a lethal complication in a liver transplant recipient. The case of a 52-year-old liver transplant recipient, who developed complete IVC as well as left iliofemoral thrombosis, is reported. After treatment with combined tissue plasminogen activator (tPA) and heparin, the IVC was successfully recanalized with sharp dissection, balloon dilatation, and stent placement. PMID- 15830292 TI - Primary malignant peripheral nerve-sheath tumor of the common bile duct. AB - Primary malignant peripheral nerve-sheath tumors of the common bile duct are extremely rare. To our knowledge, the published literature contains no previous case report of this disease. Here we report on a 58-year-old Japanese woman with a primary malignant peripheral nerve-sheath tumor of the common bile duct, which was completely resected. A hypoechoic mass was identified in the hepatic hilus, using ultrasonography and computed tomography. Endoscopic retrograde cholangiography revealed a smooth stricture and deviation of the common bile duct. Laparotomy exposed a firm mass around the common bile duct that had not invaded the surrounding tissues. Partial resection of the common bile duct and cholecystectomy were performed as the treatment of choice. The final histopathological diagnosis was malignant peripheral nerve-sheath tumor arising from the wall of the common bile duct. PMID- 15830293 TI - Pediatric nonalcoholic steatohepatitis associated with hypopituitarism. AB - We experienced two cases of pediatric nonalcoholic steatohepatitis (NASH) associated with hypopituitarism. The first patient was diagnosed with a craniopharyngioma at 5 years of age. After an operation to treat the condition, the patient gradually became obese, and an elevation of transaminases was observed. At 16 years of age, the patient was diagnosed as having NASH with liver cirrhosis. He was started on hormone replacement therapy; however, his insulin resistance and liver fibrosis, as evaluated by hyaluronic acid and platelet count, progressed. In addition, his hyperleptinemia continued. The second patient was diagnosed, at 10 years of age, as having pituitary dysfunction due to fetal asphyxia, and he was started on hormone replacement therapy. This patient was noted to have been obese throughout his life. He was diagnosed as having NASH with advanced fibrosis at 18 years of age. It is important for both hepatologists and endocrinologists to be aware of the association between pituitary dysfunction and NASH. PMID- 15830294 TI - A case of multiple colonic lymphangiomas causing intussusception. PMID- 15830295 TI - Self-made device for safe replacement of gastrostomy tube. PMID- 15830296 TI - Primary sclerosing cholangitis may be overestimated in Japan. PMID- 15830297 TI - Angiogenesis, which is essential for cancer growth, is a diagnostic and therapeutic target. PMID- 15830298 TI - When is the optimal timing for performing video capsule endoscopy for obscure gastrointestinal bleeding? PMID- 15830299 TI - Dietary intake as a risk factor for pancreatic cancer in Japan: high cholesterol and low vitamin C diet. PMID- 15830300 TI - [Comment on the Schmiegel et al. contribution--Z Gastroenterol 2004: 1129-1177]. PMID- 15830301 TI - [Patient compliance and efficacy of diagnostic procedures in the surveillance of colorectal cancer: experience from a cancer center]. AB - Postoperative surveillance is an important part of the curative therapy of colorectal cancer patients. The effort and effectiveness of these surveillance programs are controversially discussed. We analyzed the practiced follow-up of patients who had undergone a curative resection of colorectal cancer to demonstrate the difficulty to validate the performed surveillance program and to point out possible improvements. For a follow-up period of 37 months (median) we included 530 patients with at least one postoperative examination. 70 patients ended the follow-up prematurely - out of these 56 % quit the surveillance during the first 18 months. Another 68 patients died during the follow-up period. Cancer recurred in 28 % of the patients (n = 109 metastasis, n = 26 local recurrences, 18 patients developed a secondary cancer). 90 % of these recurrences occurred within the first three years. 3525 follow-up examinations took place within 79 months. Patient histories and physical examinations were not helpful for the diagnosis of local recurrences; neither were laboratory routine screenings meaningful. Carcinoembryonic antigen (CEA) and CA 19 - 9 tests, ultrasonographic studies, chest XD-rays and colonoscopic procedures had a higher diagnostic value on the other hand. We demonstrated the problematic nature of the evaluation of different follow-up tests concerning their validity as they were part of a complex postoperative surveillance program. It is also important to point out that the success of the postoperative surveillance depends strongly on the compliance of the patients. To increase this compliance we suggest that the follow-up of patients should be more strongly oriented towards the incidence of recurrences. PMID- 15830302 TI - Patient education in inflammatory bowel disease does not influence patients knowledge and long-term psychosocial well-being. AB - BACKGROUND AND AIMS: Patient education is accepted in many disciplines as a valid component of disease management in chronic diseases. The aim of this prospective study was to analyze the effects of an education program in patients with inflammatory bowel disease. METHODS: 145 patients with inflammatory bowel disease were prospectively included: 73 were educated in four sessions, 72 were educated after the one year evaluation period (control group). The following topics were presented: pathogenesis, diagnostic procedures, course of disease, medical and surgical treatment, nutrition, social problems and support, stress management, and coping with the disease. RESULTS: The repeated measurement two-way analysis of variances showed no effects of the patient education program on disease related knowledge, depression and quality of life. CONCLUSION: This patient education program was not able to increase disease-related knowledge or psychosocial variables in patients with IBD. However, most of the patients were very satisfied with the education program, since as judged by their own assessment it helped them to act responsibly for themselves and their disease. PMID- 15830303 TI - [Detection of subclinical and overt hepatic encephalopathy and treatment control after L-ornithine-L-aspartate medication by magnetic resonance spectroscopy ((1)H MRS)]. AB - Hepatic encephalopathy (HE) is a common problem in liver cirrhosis and is associated with typical changes of cerebral metabolite pattern observed by proton magnetic resonance spectroscopy (MRS). In HE, a reduction of the cerebral myo inositol (mI) and choline (Cho) and an increase of glutamine/glutamate (Glx) can typically be detected with this method. In the present study MRS was used to assess prospectively specific parameters of cerebral metabolism before and after 6 days of treatment with a low-protein diet and with L-ornithine-L-aspartate (LOLA). 6 patients with liver cirrhosis were included in this pilot study. According to standardized neuropsychological tests overt HE or subclinical HE was detected in all patients. All patients received a low-protein diet (< 60 g/d) and were treated additionally with LOLA (20 g QD i. v.). MRS examinations were done before and after 6 days of treatment and the results were compared with those of healthy volunteers. Before treatment mI/Cr ratios in the grey matter were reduced significantly in cirrhotic patients as compared to healthy volunteers (0.30 +/- 0.22 vs. 0.68 +/- 0.11; P = 0.028). In addition, patients showed a (non significant) reduction of the Cho/Cr-ratio (0.19 +/- 0.03 vs. 0.25 +/- 0.02; P = 0.17) and an elevated Glx/Cr-ratio (1.84 +/- 0.63 vs. 1.29 +/- 0.31; P = 0.05). After 6 days of treatment a significant increase of the Cho/Cr ratio (0.23 +/- 0.03 vs. 0.19 +/- 0.03; P = 0.028) was detectable and 5 of the 6 patients showed a (not significant) decrease of the elevated Glx/Cr ratios. After cessation of treatment an improvement in neuropsychological tests as shown by number connection testing (P = 0.046) as well as a decrease of elevated pre-treatment ammonia blood levels were noted. These findings, however, did not correlate with the Child-Pugh classification or evidence of clinical/subclinical HE. Using (1)H MRS it is possible to observe a specific pattern of cerebral metabolites in patients with overt and subclinical HE. In this pilot study a fast change of cerebral metabolite pattern after specific therapy of HE with LOLA was detected. Therefore, future studies with larger patient groups are needed to establish (1)H MRS as an objective method for detection and treatment control in overt and subclinical HE, especially when compared to commonly used parameters such as ammonia levels or standardized neuropsychological tests. PMID- 15830304 TI - [Esophageal resection for non-specific esophageal motility disorder]. AB - A 47-year-old patient presented with a history of dysphagia for solid food for almost 10 years and weight loss of more than 50 kg. Non-resecting surgical as well as endoscopic procedures (laparoscopic cardiomyotomy with secondary antireflux operations, balloon dilation, Botulinum-toxin injection) were without success. A barium esophagogram showed a confinement of the distal esophagus with a filiform passage of the contrast medium and undigested food in the prestenotic dilated esophageal corpus. Manometry displayed a hypertensive lower esophageal sphincter with a resting pressure of 43.8 mmHg - although completely relaxing. The tubular esophagus was aperistaltic with 100 % simultaneous and repetitive contractions. As all attempts of previous therapy had failed, a transhiatal esophagectomy with gastric pull-up and cervical esophagogastrostomy ensued. Neuropathological examination of the esophagus showed that degeneration of the myenteric plexus was not severely involved, whereas inflammatory and fibrotic changes were obvious. Esophageal resection provided the only chance of a long term benefit for our patient with relief of dysphagia. PMID- 15830305 TI - [Differencial diagnosis of gastroesophageal reflux disease -- eosinophilic esophagitis: case report]. AB - We report on a 22-year-old man with dysphagia and repeated bolus impaction in the esophagus for 10 years. Bolus impactions were frequently mobilised using an endoscope. At endoscopy, esophagitis IV degrees was described. After treatment with omeprazol there was no improvement. The patient was submitted to our hospital for fundoplication. pH-metry demonstrated an increased reflux. At endoscopy of the esophagus, we found red stripes which did not show the typical appearance of erosions. Manometry and X-ray films of the esophagus did not reveal any pathological findings. In combination with anamnesis, symptoms, and endoscopy, the diagnosis of eosinophilic esophagitis was documented by histology. After administration of oral corticosteroids a rapid improvement of the clinical symptoms was observed. The diagnosis of eosinophilic esophagitis should be kept in mind in patients with chronic symptoms of gastroesophageal reflux persisting despite medical therapy, pathological pH-metry and repeated bolus impactions. PMID- 15830306 TI - [Antiemetic prophylaxis in the chemotherapy of gastrointestinal tumours]. AB - Systemic treatment options in gastrointestinal malignancies have increased markedly. At the same time, the need for supportive measures has become more complex. Nausea and vomiting continue to impair the patients' quality of life and to jeopardise the goals of chemotherapy. Antiemetic strategies as proposed by treatment guidelines should be employed consistently in daily clinical practice. Deficits in cancer care exist in this area. In addition, newly available antiemetic drugs should be considered. Aprepitant is the first approved representative of a new drug class. Aprepitant inhibits substance P binding to the neurokinin-1-receptor. Given orally on the first three days of a cisplatin based chemotherapy in combination with a standard antiemetic regimen, aprepitant proved to be significantly more effective in the prevention of nausea and vomiting compared to the standard regimen without aprepitant. Recently presented results for chemotherapy with moderate emetogenic risks indicate that aprepitant shows superior effectiveness even in this setting. Palonosetron is a new drug in the class of 5-HT (3) (serotonin) receptor antagonists. Compared to older setrones, palonosetron exhibits a higher receptor binding activity, a longer half life, and a slightly improved activity in the prevention of nausea and vomiting after chemotherapy with moderate emetogenic risks. The implementation of standardised treatment guidelines into clinical practice will contribute to a higher patient satisfaction and a more effective utilisation of economic resources. PMID- 15830307 TI - [Esophageal carcinoma -- current status in diagnosis and therapy]. AB - Esophageal carcinoma is characterized by an increasing incidence in the last 15 years whereas the prognosis still remains almost invariably poor. The therapy of choice is still deemed to be surgical resection including extended lymphadenectomy which is accompanied by considerable amounts of peri- and postoperative complications and only rare long-time survival. Precursor lesions and early stage cancers are therefore increasingly often treated by local ablative procedures including mucosectomy, which demonstrates promising results. In advanced tumor stages definitive chemoradiation provided long-term survival equivalent to that in contemporary surgical series and thus represents another alternative to tumor resection. This review points out the epidemiology, aetiology, staging and consequences in therapy for esophageal carcinoma considering current points of view. PMID- 15830308 TI - Konjetzny: a German surgeon of the past century and his pioneering hypothesis of a bacterial aetiology for gastritis, peptic ulcer and gastric cancer. PMID- 15830311 TI - [Is asynchronous ventricular pacemaker stimulation dangerous? Results of an international survey]. AB - BACKGROUND: The safety of asynchronous ventricular pacemaker stimulation, occurring with magnet application or during noise reversion, is a point of controversial discussions. A survey was performed, to evaluate such complications. METHODS: 145 pacemaker specialists worldwide were asked about their pacemaker experience and arrhythmia induction due to asynchronous stimulation. RESULTS: 102/145 physicians (70 %) answered. Pacing experience: 3 - 40 y, mean 20 +/- 8 y, cumulative 1,974 y. Pacemaker follow-ups: 50 - 12,000/y, mean 2,326 +/- 2,082, overall 230,305/y. No data: 3. Arrhythmia induction was reported by 34/102 physicians (33 %) in 48 cases. 23 cases were major complications: In 13 pts VT or VF was induced. 4 of these had an ejection fraction < 30 %, one patient died. 10 additional cases without detailed information were included. 4 of these occurred 15 to 20 years ago. No case occurred in the last 7 years. 25 events were induced secondary, or clinically not relevant: 14 cases had asymptomatic PVCs or short runs. 5 cases 20 - 25 years ago, were due to anodal stimulation. 5 events happened during acute myocardial ischemia or electrolyte imbalance immediately after cardiac surgery. One patient had spontaneous VTs as well. CONCLUSION: Inducement of ventricular tachyarrhythmias during asynchronous ventricular pacing is an extreme rarity. Patients at risk for these rare events are only those with acute myocardial ischemia or severe electrolyte imbalance, and those with organic cardiac disease, reduced left ventricular function and spontaneous ventricular tachyarrhythmias. Asynchronous ventricular pacing can be considered as safe. PMID- 15830312 TI - [Systemic fibrinolytic therapy using urokinase in central retinal artery occlusion. A case study]. AB - BACKGROUND AND OBJECTIVE: Systemic fibrinolysis has become an important therapeutical option in patients with thrombotic occlusion of coronary or pulmonary arteries. In view of the hemorrhagic risk systemic fibrinolytic therapy for retinal vessel occlusion has been discussed controversial. In the present case study results and complications of systemic fibrinolysis should be investigated in patients with central retinal artery occlusion. PATIENTS AND METHODS: From 1995 to 2002 a case series of 19 consecutive patients (8 female, 11 male, age: 63.2+/-14,3 years) with central retinal artery occlusion were treated by systemic application of urokinase using a standardized scheme. The latency from initial symptoms until the initiation of therapy and the medical history of the patients were documented. Visual acuity was determined on admission and before discharge and possible complications were documented. Additionally, screening investigations for genetic thrombophilia were performed. RESULTS: 15 patients showed an improvement of the visual acuity (79 %, 95 %-KI: 54 %-94 %). For 3 patients no improvement and for one patient a decrease of the visual acuity was determined. Hemorrhagic complications were observed in two patients (11 %, 95 %-KI: 1 %-33 %). As these minor bleedings were self-limiting the fibrinolytic therapy was discontinued only in one patient. As risk factors most commonly arterial hypertension (68 %) and smoking (26 %) were identified. In 4 patients a genetic thrombophilia was diagnosed. CONCLUSIONS: Considering the poor prognosis of central retinal artery occlusion and the disappointing results of conservative treatment, an improvement of the visual acuity in the absence of critical complications was observed with systemic fibrinolytic therapy in the presented case study. However, only controlled trials can provide proof for the effect of fibrinolysis versus spontaneous improvement. PMID- 15830314 TI - [Postoperative arrhythmias]. PMID- 15830313 TI - [Recurrent strongyloidiasis as an indicator of HTLV-1 infection]. AB - HISTORY AND CLINICAL FINDINGS: A 53-year-old West African man presented two years after a travel to Guinea because of severe headache, neck stiffnes, fever and pruritus. The patient had been in orthopedical treatment for the last five months. INVESTIGATIONS: Stool microscopy revealed a high number of Strongyloides stercoralis larvae. Hematology, biochemistry and all other parasitology results were normal. HIV-1/2 testing was negative and CD4+-lymphocyte count was normal. Concomitant infection by Human T Cell lymphotropic virus type 1 (HTLV-1) was confirmed by serology and PCR. The phylogenetic analysis confirmed African origin of the virus. TREATMENT: The infection responded to a five-day course of albendazol at 400 mg/d but during the following five years repeat recrudescences were observed inspite of high-dosage and prolonged antiparasitic treatments. Eventually, eradication of the infection was achieved by a four day course of ivermectin 0.2 mg/kg/d. CONCLUSIONS: Although both strongyloidiasis and HTLV-1 infections occur most frequently in tropical areas, these may also be observed in temperate regions. Suppression of the immune system by HTLV-1 differs from that by HIV. CD4+-lymphocytes were rarely decreased. Prolonged treatment with ivermectin in a dosage exceeding the current recommendations may be required in HTLV-1 infected patients and was well tolerated. The unusual presentation of the infection with muscular symptoms contributed to the delay of the diagnosis. HTLV 1 positive patients must be monitored for years. They and their partners must be instructed how to prevent transmission of the virus. PMID- 15830315 TI - [Mutations of the gene coding for the receptor tyrosine kinase FLT3 in acute myeloid leukemia. Significance as the disease-specific molecular marker for diagnosis, prognosis and innovative therapy approaches]. PMID- 15830316 TI - [Gastrointestinal bleeding during low dose acetylsalicylic acid. Is Godamed-100 TAH an alternative to conventional ASS preparations?]. PMID- 15830317 TI - [Critical care medicine: who is allowed it, who needs it? Comment on the article from DMW 1/7/2005]. PMID- 15830319 TI - [Critical care medicine: who is allowed it, who needs it? Comment on article from DMW 1/7/2005]. PMID- 15830321 TI - [Antibiotic use in Germany in comparison with Europe. Comment on the article from DMW 38/2004]. PMID- 15830323 TI - Current strategies for articular cartilage repair. AB - Defects of articular cartilage that do not penetrate to the subchondral bone fail to heal spontaneously. Defects that penetrate to the subchondral bone elicit an intrinsic repair response that yields a fibrocartilaginous repair tissue which is a poor substitute for hyaline articular cartilage. Many arthroscopic repair strategies employed utilise this intrinsic repair response to induce the formation of a repair tissue within the defect. The goal, however, is to produce a repair tissue that has the same functional and mechanical properties of hyaline articular cartilage. To this end, autologous osteochondral transfer can provide symptomatic relief. This technique involves the excision of healthy cartilage plugs from 'non-load bearing' regions of the joint for implantation into the defect. Cell based transplantation methods currently involve the transplantation of expanded autologous chondrocytes to the defects to form a repair tissue. This technique again involves the excision of healthy cartilage from the joint for expansion. Current research is exploring the potential use of mesenchymal stem cells as a source for tissue engineering, as well as the combination of cells with biodegradable scaffolds. Although current repair strategies improve joint function, further research is required to prevent future degeneration of repair tissue. PMID- 15830322 TI - Candidate-gene screening and association analysis at the autism-susceptibility locus on chromosome 16p: evidence of association at GRIN2A and ABAT. AB - Autism is a highly heritable neurodevelopmental disorder whose underlying genetic causes have yet to be identified. To date, there have been eight genome screens for autism, two of which identified a putative susceptibility locus on chromosome 16p. In the present study, 10 positional candidate genes that map to 16p11-13 were examined for coding variants: A2BP1, ABAT, BFAR, CREBBP, EMP2, GRIN2A, MRTF B, SSTR5, TBX6, and UBN1. Screening of all coding and regulatory regions by denaturing high-performance liquid chromatography identified seven nonsynonymous changes. Five of these mutations were found to cosegregate with autism, but the mutations are not predicted to have deleterious effects on protein structure and are unlikely to represent significant etiological variants. Selected variants from candidate genes were genotyped in the entire International Molecular Genetics Study of Autism Consortium collection of 239 multiplex families and were tested for association with autism by use of the pedigree disequilibrium test. Additionally, genotype frequencies were compared between 239 unrelated affected individuals and 192 controls. Patterns of linkage disequilibrium were investigated, and the transmission of haplotypes across candidate genes was tested for association. Evidence of single-marker association was found for variants in ABAT, CREBBP, and GRIN2A. Within these genes, 12 single-nucleotide polymorphisms (SNPs) were subsequently genotyped in 91 autism trios (one affected individual and two unaffected parents), and the association was replicated within GRIN2A (Fisher's exact test, P<.0001). Logistic regression analysis of SNP data across GRIN2A and ABAT showed a trend toward haplotypic differences between cases and controls. PMID- 15830325 TI - Proceedings of a consensus conference: towards an understanding of TRALI. AB - Transfusion-related acute lung injury is a relatively uncommon transfusion associated adverse effect occurring during or soon after an allogeneic blood transfusion. Transfusion-related acute lung injury is a complex syndrome that has many manifestations and has only recently been identified to be an important cause of transfusion-associated morbidity and mortality. But despite its increasing recognition, much about the pathogenesis, treatment, and prevention is poorly understood and often controversial. The purpose of this consensus conference was to bring together international experts in an effort to try to standardize a case definition, which could be used to enhance future understanding of transfusion-related acute lung injury including its epidemiology, pathogenesis, management, prevention, and research. These proceedings are being provided with a view to making available to the transfusion medicine community the considerable amount of important information presented at this consensus conference by the invited international panel of experts. PMID- 15830326 TI - Erythrocyte adhesion receptors: blood group antigens and related molecules. AB - During the second half of the 20th century, blood bankers quickly expanded our knowledge of human erythrocyte blood group antigens. By the dawn of the 21st century, several hundred blood group antigen polymorphisms had been identified. Hot on the heels of the serologists, membrane biochemists and molecular geneticists defined both the biochemical and genetic bases of most of these antigens. Perhaps to their surprise, this work has led to the discovery of functionally diverse and important membrane proteins expressed on the surface of red cells, including numerous adhesion molecules. Red cells express an unexpected number of such adhesion receptors, some of which contribute to human disease, as well as to normal red cell development. And perhaps most interestingly, study of these molecules has elucidated ways in which even mature red cells respond to external stimuli, such as adrenergic hormones. PMID- 15830327 TI - Human blood group genes 2004: chromosomal locations and cloning strategies. AB - Of the 29 human blood group system genes, 27 have been localized to 14 autosomes and 2 have been assigned to the X chromosome. It is remarkable that 28 of the 29 system genes have now been localized to a single cytogenetic band on a specific chromosome. In this review, we summarize the chromosomal locations and cloning strategies used for those genes encoding blood group systems. We highlight such information about the 3 most recently defined blood group systems (I, GLOB, and GIL). In addition, we provide new information about 2 older blood group systems (SC and RAPH) whose polymorphisms have been defined in cloned genes. PMID- 15830328 TI - Can we improve the management of blood donors with nonspecific reactivity in viral screening and confirmatory assays? AB - Donors with nonspecific reactivity in viral screening or confirmatory assays are problematic for blood services because of donor management issues and product loss. Considerable experience has now accumulated in the use of screening and confirmatory assays; therefore, it is timely to examine the ways in which donors with nonspecific reactivity are managed. In this review, we summarize the causes and characteristics of nonspecific reactivity in blood donors and approaches for reducing the number of nonspecific reactive results and we offer some suggestions for improving the management of these donors. PMID- 15830329 TI - The cost of blood: multidisciplinary consensus conference for a standard methodology. AB - Prior attempts to account for the cost of blood have varied in economic perspective, methodology, and scope and may have underestimated both direct and indirect costs associated with transfusions. To devise a comprehensive and standardized methodology for the United States that will improve upon existing estimates, a panel of experts in blood banking and transfusion medicine was assembled and participated in consensus deliberations using modified Delphi methods. As a first step, a process-flow model that describes all the major steps involved in collecting, processing, and transfusing blood such as donor recruitment and follow-up of transfusion sequelae was constructed. Next, interdependencies were outlined and detailed cost elements within each step were itemized. The relative importance of each element was rated. Personnel, screening for infectious agents, information systems, laboratory evaluations, management of transfusion reactions, and equipment were ranked as the most important factors to capture but, in an effort to be all-inclusive, even minor elements were included. This consensus model is broad-based and should serve societal, provider, and payer perspectives for future cost studies. Recognizing the limitations of process-flow models, the next iteration will use an activity-based approach to more fully account for the cost of blood than present estimates. PMID- 15830330 TI - Putting the pieces together: Roger I. Lee and modern transfusion medicine. AB - Roger Irving Lee (1881-1964) played significant scientific and leadership roles in overcoming the clinical impediments to blood transfusion in the 1910s. He developed the first successful anticoagulant system, paraffinized glass, and the first sterile system for indirect transfusion without defibrination, the Lee Vincent flask. He used citrate as an anticoagulant before those generally credited with its discovery did. He introduced surgical antisepsis of the donor site and a practical system for maintaining typed blood for a large hospital donor and transfusion service. He was Oswald Robertson's mentor at Harvard as well as his commanding officer in the Harvard Medical Unit on the Western Front during World War I. Lee sent Robertson to perform "his preserved blood cells transfusion" in the casualty clearing stations of the British Third Army and provided Robertson's group with O donors, enabling the most important medical development of the war. Lee put the pieces of modern blood banking and transfusion together. PMID- 15830332 TI - Peptide-polymer hybrid nanotubes. PMID- 15830333 TI - Molecular mechanisms of the chemopreventive effects of resveratrol and its analogs in carcinogenesis. AB - Resveratrol (3,4',5-trihydroxy-trans-stilbene), a phytoalexin found in grape skins, peanuts, and red wine, has been reported to exhibit a wide range of biological and pharmacological properties. It has been speculated that dietary resveratrol could be an explanation for the so-called 'French paradox' as it may act as an antioxidant, promote nitric oxide production, inhibit platelet aggregation, and increase high-density lipoprotein cholesterol, and thereby serve as a cardioprotective agent. Recently, it has been demonstrated that resveratrol can function as a cancer chemopreventive agent, and there has been a great deal of experimental effort directed toward defining this effect. It has been shown that resveratrol and some of its analogs interfere with signal transduction pathways, modulate cell cycle-regulating proteins, and is a potent inducer of apoptosis in multiple carcinoma cell lines. This review summarizes the recent advances that have provided new insights into the molecular mechanisms underlying the promising properties of resveratrol. PMID- 15830334 TI - Resveratrol: preventing properties against vascular alterations and ageing. AB - Cardiovascular diseases are the leading cause of death in developed countries where the common pathological substrate underlying this process is atherosclerosis. Several new concepts have emerged in relation to mechanisms that contribute to the regulation of the vascular diseases and associated inflammatory effects. Recently, potential antioxidants (vitamin E, polyphenols) have received much attention as potential anti-atherosclerotic agents. Among the polyphenols with health benefic properties, resveratrol, a phytoalexin of grape, seem to be a good candidate protecting the vascular walls from oxidation, inflammation, platelet aggregation, and thrombus formation. In this review, we focus on the mechanism of resveratrol cardiovascular benefic effects. We analyze, in relation with the different steps of atherosclerotic process, the resveratrol properties at multiple levels, such as cellular signaling, enzymatic pathways, apoptosis, and gene expression. We show and discuss the relationship with reactive oxygen species, regulation of pro-inflammatory genes including cycloxygenases and cytokines in molecular inflammatory and aging processes, and how the regulation of these activites by resveratrol can lead to a prevention of vascular diseases. PMID- 15830335 TI - Dietary grape supplement ameliorates cerebral ischemia-induced neuronal death in gerbils. AB - Oxidative damage has been implicated as one of the leading causes for neuronal cell death in a number of neurodegenerative diseases including stroke. Many vegetables and fruits are enriched in polyphenolic compounds known to exhibit antioxidant properties. This study is to investigate whether dietary supplement with grape powder (GP) may offer protection against neuronal damage due to global cerebral ischemia induced to Mongolian gerbils by occlusion of the common carotid arteries, a model known to cause delayed neuronal death (DND) in the hippocampal CA1 area. Gerbils were fed either a control diet (AIN76a) or a control diet supplemented with low (5.0 g/kg diet) or high (50 g/kg diet) levels of GP for two months. Four days after ischemia/reperfusion (I/R), the extent of DND, glial cell activation, nuclear DNA oxidation, and apoptotic terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) reaction in hippocampal CA1 region were assessed. Ischemia-induced extensive DND in the CA1 region was accompanied by oxidative and fragmented DNA damage and a marked increase in reactive astrocytes and microglial cells. Dietary GP supplementation significantly protected neurons against I/R-induced DND, DNA damage, and apoptosis as well as attenuated glial cell activation. These results demonstrate that due to the antioxidant properties of polyphenols in GP, nutritional diets supplemented with grape can protect the brain against ischemic damage. The neuroprotective effects of GP supplement may have wide implication in the future for prevention/protection against other neurodegenerative damage. PMID- 15830336 TI - Bioavailability of trans-resveratrol from red wine in humans. AB - Many in vitro studies demonstrated significant biological effects of trans resveratrol. Thus, understanding the rate of intestinal absorption and metabolization in vivo of trans-resveratrol is the prerequisite to evaluate its potential health impact. Bioavailability studies mainly in animals or in humans using the pure compound at very high doses were performed. In this work, trans resveratrol bioavailability from a moderate consumption of red wine in 25 healthy humans has been studied by three different experiments. The wine ingestion was associated to three different dietary approaches: fasting, a standard meal, a meal with high and low amount of lipids. Trans-resveratrol 3- and 4'-glucuronides were synthesized, purified, and characterized as pure standards. Bioavailability data were obtained by measuring the concentration of free, 3-glucuronide and 4' glucuronide trans-resveratrol by high-performance liquid chromatography (HPLC), both with ultraviolet (UV) and mass spectrometry (MS) detection, in serum samples taken at different times after red wine administration. Free trans-resveratrol was found, in trace amounts, only in some serum samples collected 30 min after red wine ingestion while after longer times resveratrol glucuronides predominated. Trans-resveratrol bioavailability was shown to be independent from the meal or its lipid content. The finding in human serum of trans-resveratrol glucuronides, rather than the free form of the compound, with a high interindividual variability, raises some doubts about the health effects of dietary resveratrol consumption and suggests that the benefits associated to red wine consumption could be probably due to the whole antioxidant pool present in red wine. PMID- 15830337 TI - Maillard reaction products as "natural antibrowning" agents in fruit and vegetable technology. AB - The effects of Maillard reaction products (MRPs), synthesized from a sugar (pentose, hexose, or disaccharide) and either a cysteine-related compound, an amino acid, or a sulfur compound, were investigated on polyphenoloxidase (PPO) activity from apple, mushroom, and eggplant. The optimal conditions for the production of inhibitory MRPs were performed using two-factor and five-level central experimental designs. It resulted that thiol-derived MRPs were highly prone to give rise to inhibitory compounds of PPO activity. Technological assays were also performed to test the efficiency of selected MRPs in the prevention of enzymatic browning in raw and minimally processed fruits and vegetables. PMID- 15830338 TI - Trans- and cis-resveratrol increase cytoplasmic calcium levels in A7r5 vascular smooth muscle cells. AB - The effects of trans- and cis-resveratrol on cytosolic Ca2+ concentration ([Ca2+]i) were studied using fura-2 in vascular smooth muscle cells (A7r5). Both isomers of resveratrol caused a sustained elevation in [Ca2+]i, cis-resveratrol being significantly more effective than the trans-isomer. The resveratrol-induced increase in [Ca2+]i was significantly potentiated by the previous application of low concentrations of thapsigargin, partially inhibited by nifedipine or Ni2+, and not affected by SKF 96365. In the absence of extracellular Ca2+, both isomers of resveratrol induced a transient, slow increase in [Ca2+]i, which was inhibited by the previous depletion of intracellular stores with thapsigargin and completely blocked by preincubation with TMB-8, an inhibitor of intracellular calcium release. Reintroduction of Ca2+ in the external solution after the resveratrol-induced release of Ca2+ activated the Ca2+ influx through store operated calcium channels. The resveratrol-induced increase in [Ca2+]i in the absence of extracelullar Ca2+ partially reduced the increase in [Ca2+]i evoked by the subsequent application of thapsigargin. Our results suggest that trans- and cis-resveratrol induce a depletion of Ca2+ from the same intracellular stores released by thapsigargin and subsequent capacitative influx of Ca2+. Additionally, a direct activation of transmembrane Ca2+ influx through another type of channel may be also implicated. PMID- 15830339 TI - Management of suspicious or indeterminate calcifications and impact on local control. AB - BACKGROUND: The current study was undertaken to determine the potential advantage associated with complete removal of suspicious or indeterminate calcifications (SIC) before the initiation of irradiation as part of breast conservation therapy (BCT). METHODS: Of 2045 patients treated with BCT at Yale University School of Medicine (New Haven, Connecticut) and satellite facilities before 2002, 111 women, included 3 patients with bilateral disease, had a postexcision preirradiation mammogram (PREMAMMO) to evaluate residual SIC. Thus, 114 breasts were at risk for local disease recurrence. Seventy-five breasts at risk had no residual SIC and proceeded to undergo radiotherapy (XRT) without further surgery or mammography. Of the remaining 39 breasts at risk, only 3 underwent a PREMAMMO with documented removal of all calcifications (DRC). Thirty-six breasts at risk proceeded to XRT with either known SIC or with nondocumented removal of calcifications (NDRC) after another excision. RESULTS: Of the 78 breasts at risk with DRC via PREMAMMO, there were 7 local failures (LF) and 1 distant failure. Of the 36 breasts with NDRC via PREMAMMO, there were 7 LF and 1 regional failure. Of the 34 breasts who underwent reexcision after detection of SIC by PREMAMMO, 20 (59%) were found to have residual disease. CONCLUSIONS: Patients with DRC were found to have better local control than patients with NDRC. In addition, the presence of SIC on a PREMAMMO was associated with a high probability of detecting residual disease. PMID- 15830340 TI - Large-scale gene expression analysis of cholesterol dependence in NS0 cells. AB - NS0, a nonsecreting mouse myeloma cell, is a major host line used for recombinant antibody production. These cells have a cholesterol-dependent phenotype and rely on an exogenous supply of cholesterol for their survival and growth. To better understand the physiology underlying cholesterol dependence, we compared NS0 cells, cultivated under standard cholesterol-dependent growth conditions (NS0), to cells adapted to cholesterol-independent conditions (NS0 revertant, NS0_r). Large-scale transcriptional analyses were done using the Affymetrix GeneChip array, MG-U74Av2. The transcripts expressed differentially across the two cell lines were identified. Additionally, proteomic tools were employed to analyze cell lysates from these two cell lines. Cellular proteins from both NS0 and NS0_r were subjected to 2D gel electrophoresis. MALDI-TOF mass spectrometry was performed to determine the identity of the differentially expressed spots. We examined the expression level of mouse genes directly involved in cholesterol biosynthesis, lipid metabolism, and central energy metabolism. Most of these genes were downregulated in the revertant cell type, NS0_r, compared to NS0. Overall, a large number of genes are expressed differentially, indicating that the reversal of cholesterol dependency has a profound effect on cell physiology. It is probable that a single gene mutation, activation, or inactivation is responsible for cholesterol auxotrophy. However, the wide-ranging changes in gene expression point to the distinct possibility of a regulatory event affecting the reversibility of auxotrophy, either directly or indirectly. PMID- 15830341 TI - Expression and structure of interleukin 4 receptors in primary meningeal tumors. AB - BACKGROUND: It was reported previously that malignant human tumors, like glioma and medulloblastoma, express high-density interleukin (IL-4) receptor mRNA and protein. Because IL-4 receptors (R) are sensitive targets for targeted therapeutics, knowledge of the expression of these receptors in other central nervous system tumors is of great interest. In this study, the authors examined the expression and subunit composition of IL-4R complex in primary human meningiomas. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) analysis for IL-13Ralpha1, IL-4Ralpha and IL-2Rgammac was performed on total RNA extracted from 35 meningiomas and a normal human brain tissue sample. Results were confirmed in nine randomly selected tumors by quantitative real-time PCR and in situ immunofluorescence assay. RESULTS: Transcripts for the IL-4Ralpha and IL 13Ralpha1 chains were overexpressed in meningiomas compared with normal brain tissue. The levels of IL-4Ralpha mRNA appeared to be higher compared with the levels of IL-13Ralpha1 mRNA. The results also showed that tumors with higher disease grade tended to have increased mRNA expression for the IL-4Ralpha chain. This IL-4Ralpha mRNA overexpression appeared to be more frequent in younger patients (age < 37 years). The transcripts for IL-2Rgammac chain were not detected in any of the tumor samples or in normal brain tissue. Quantitative real time PCR confirmed the results of the RT-PCR analysis. Meningiomas also demonstrated a bright immunofluorescent staining for the IL-4Ralpha and IL 13Ralpha1 chains but no staining for IL-2Rgammac. CONCLUSIONS: Expression of the IL-4Ralpha and IL-13Ralpha1 chains and absence of IL-2gammac expression established that meningiomas expressed type II IL-4Rs. These receptors may serve as a target for cytotoxin/immunotoxin therapy in patients with meningioma who are not amenable to surgical resection or for recurrent tumors. PMID- 15830342 TI - Modulation of tamoxifen sensitivity by antisense Bcl-2 and trastuzumab in breast carcinoma cells. AB - BACKGROUND: Because the overexpression of HER-2 and Bcl-2 is associated with resistance to tamoxifen (TAM), the authors examined the effect of antisense (AS) Bcl-2 on sensitivity to TAM compared with the effect of trastuzumab on sensitivity to TAM in breast carcinoma cell lines. METHODS: Drug sensitivity was assessed in vitro using a [3-4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay with the breast carcinoma cell lines ZR-75-1, MDA-MB-453, and BT 474. AS Bcl-2 18-mer phosphorothioate oligonucleotide was applied. Apoptotic cell death was assessed with the terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick-end labeling method, and gene expression was evaluated with Western blot analysis. RESULTS: The expression of Bcl-2 was identified in ZR-75-1 and BT-474 cells and, to a lesser extent, in MDA-MB-453 cells. Overexpression of HER-2 was identified in BT-474 cells, and moderate expression was identified in MDA-MB-453 and ZR-75-1 cells. Combination treatment with trastuzumab or AS Bcl-2 enhanced TAM sensitivity in ZR-75-1 cells, which showed 50% inhibitory concentration (IC50) values of 0.9 microM (7.2-fold increase) and 0.5 microM (13.0-fold), respectively. Combination treatment with trastuzumab or AS Bcl-2 slightly enhanced TAM sensitivity of BT-474 cells, with IC50 values of 3.0 microM (1.3-fold) and 1.5 microM (2.6-fold), respectively. The sensitivity of MDA-MB-453 cells to TAM was not enhanced by combination with trastuzumab or AS Bcl-2. Modulation of TAM sensitivity by AS Bcl-2 was superior to modulation by trastuzumab in HER-2-expressing and Bcl-2-expressing breast carcinoma cells. Enhanced sensitivity in combination with AS Bcl-2 was associated with down regulation of Bcl-2 and pAkt, which was correlated with the induction of Bax and caspase-3, leading to apoptosis. CONCLUSIONS: AS Bcl-2 appeared to be superior to trastuzumab with respect to regulating the signal-transduction pathways involved in breast carcinoma cells. PMID- 15830343 TI - Architecture of the ciliary muscle of Gallus domesticus. AB - There are species-related anatomical differences in the ciliary muscle of the avian eye. The arrangement of muscular fibers in the avian eye is not well defined. To clarify this situation, we studied the architecture of ciliary muscle of Gallus domesticus by light and scanning electron microscopy (SEM). Our results showed the existence of three main muscular groups that we defined as anterior, posterior, and intermediary. These muscle divisions correspond to the description of the ciliary muscle as previously stated by Crampton (1813), Brucke, and Muller (1856). The striated fibers have a meridian orientation. The anterior and posterior muscular groups are inserted in the sclera, around the Schlemm's canal wall and ciliary process stroma. The vitreal intermediary muscle has fibers inserted in Schlemm's canal wall and ciliary process stroma. The framework of these muscular fibers may according to its insertions participate in the visual accommodation mechanism and outflow of the aqueous humor system. PMID- 15830344 TI - Energizing cell-free protein synthesis with glucose metabolism. AB - In traditional cell-free protein synthesis reactions, the energy source (typically phosphoenolpyruvate (PEP) or creatine phosphate) is the most expensive substrate. However, for most biotechnology applications glucose is the preferred commercial substrate. Previous attempts to use glucose in cell-free protein synthesis reactions have been unsuccessful. We have now developed a cell-free protein synthesis reaction where PEP is replaced by either glucose or glucose-6 phosphate (G6P) as the energy source, thus allowing these reactions to compete more effectively with in vivo protein production technologies. We demonstrate high protein yields in a simple batch-format reaction through pH control and alleviation of phosphate limitation. G6P reactions can produce high protein levels ( approximately 700 microg/mL of chloramphenical acetyl transferase (CAT)) when pH is stabilized through replacement of the HEPES buffer with Bis-Tris. Protein synthesis with glucose as an energy source is also possible, and CAT yields of approximately 550 mug/mL are seen when both 10 mM phosphate is added to alleviate phosphate limitations and the Bis-Tris buffer concentration is increased to stabilize pH. By following radioactivity from [U-(14)C]-glucose, we find that glucose is primarily metabolized to the anaerobic products, acetate and lactate. The ability to use glucose as an energy source in cell-free reactions is important not only for inexpensive ATP generation during protein synthesis, but also as an example of how complex biological systems can be understood and exploited through cell-free biology. PMID- 15830345 TI - Survival benefit with imatinib mesylate therapy in patients with accelerated phase chronic myelogenous leukemia--comparison with historic experience. AB - BACKGROUND: The effect of imatinib mesylate on survival in the accelerated phase of chronic myelogenous leukemia (CML) is unknown. The objectives of this study were to update the long-term experience with imatinib in patients who had accelerated-phase CML and to compare outcomes with historic experience. METHODS: The outcomes of 176 patients who received treatment with imatinib were reviewed and compared with the outcomes of 213 historic control patients with accelerated phase CML who received treatment with interferon-alpha or with other modalities. RESULTS: With imatinib, the complete hematologic response rate was 82% versus a rate < or = 50% for others, and the complete cytogenetic response rate was 43% versus rates of 0-6% for others. The estimated 4-year survival rates were 53% with imatinib, 42% with interferon-alpha, and 0-21% for others. A multivariate analysis of the total population of 389 patients indicated that imatinib therapy (vs. other therapies) was an independent, favorable prognostic factor for survival (P < 0.0001; hazard rate, 0.62). A subset analysis that included only patients who were treated with imatinib and interferon-alpha (276 patients) also identified imatinib as an independent favorable prognostic factor (P < 0.0001; hazard rate, 0.65). The 3-month cytogenetic response to imatinib was associated with significantly different survival outcomes (P < 0.0001). A multivariate analysis that included pretreatment characteristics and 3-month cytogenetic response among 150 patients who received imatinib and were alive at 3 months identified only 2 adverse independent prognostic factors: lack of a cytogenetic response at 3 months (P < 0.001) and anemia (hemoglobin < 10 g/dL; P = 0.003). Patients who had neither factor (41%) had an estimated 4-year survival rate of 88%; in the other patients, the 4-year survival rate was < or = 60%. This may have implications in relation to subsequent therapy, because, according to the outcomes of patients who underwent allogeneic transplantation in accelerated phase at the authors' institution and from literature reports, the estimates of 5 year survival were 25-30%. CONCLUSIONS: The current results suggest that imatinib improved survival compared with other therapies in patients with accelerated phase CML. PMID- 15830346 TI - Results of a phase I dose-escalation study using three-dimensional conformal radiotherapy in the treatment of inoperable nonsmall cell lung carcinoma. AB - BACKGROUND: The objective of this study was to report the results of a Phase I dose-escalation study using three-dimensional conformal radiation therapy (3D CRT) for the treatment of patients with nonsmall cell lung carcinoma (NSCLC). METHODS: Between 1991 and 2003, 104 patients were enrolled for 3D-CRT at Memorial Sloan-Kettering Cancer Center. The median patient age was 69 years. Twenty-eight percent of patients had Stage I-II NSCLC, 33% of patients had Stage IIIA NSCLC, 32% of patients had Stage IIIB NSCLC, and 6% of patients had recurrent NSCLC. Induction chemotherapy was received by 16% of patients. Radiation was delivered in daily fractions of 1.8 grays (Gy) for doses < or = 81.0 Gy and in daily fractions of 2.0 Gy for higher doses. Accrual at a dose level was complete when 10 patients received the intended dose without unacceptable acute morbidity. RESULTS: After an incident of fatal acute radiation pneumonitis at the starting dose of 70.2 Gy, the protocol was modified to limit normal tissue complication probabilities (NTCP) to < 25%. The dose was then escalated from 70.2 Gy, to 75.6 Gy, 81.0 Gy, and 84.0 Gy, with at least 10 patients treated at each dose level. Unacceptable pulmonary toxicity occurred at 90.0 Gy. Subsequently, another 10 patients were accrued at the 84.0 Gy level with acceptable toxicity. Thus, 84.0 Gy was the maximum tolerated dose (MTD). The crude late pulmonary toxicity rate was 7%, the 2-year local control rate was 52%, the disease-free survival rate was 33%, and the overall survival rate was 40%. The median survival was 21.1 months. Overall survival was improved significantly in patients who received > or = 80.0 Gy. CONCLUSIONS: The MTD of 3D-CRT for NSCLC with an NTCP constraint of 25% was 84.0 Gy in the current study. There was a suggestion of improved survival in patients who received 80.0 Gy. PMID- 15830347 TI - Factors affecting bone mineral density in patients with prostate carcinoma before and after orchidectomy. AB - BACKGROUND: Orchidectomy is an accepted form of androgen-deprivation therapy (ADT) for prostate carcinoma. Osteoporosis is common in elderly individuals and is accelerated by ADT. The authors studied changes in bone mineral density (BMD) after ADT and factors that affected those changes. METHODS: Fifty patients with prostatic adenocarcinoma who opted to undergo orchidectomy were studied prospectively. All patients completed 6 months of follow-up, and 20 of those patients completed 12 months of follow-up. Patients' age, weight, height, body mass index (BMI), physical activity, addiction (smoking, alcohol), dietary calcium intake, and lactose tolerance status were noted. Lumbar spinal (L1-L3) trabecular BMD was measured with quantitative computed tomography (QCT) at baseline and every 6 months for 1 year and was compared with preoperative values. The effects of various patient characteristics on preoperative BMD and changes in BMD also were analyzed. RESULTS: The mean +/- standard deviation (SD) age of the patients was 69.5 +/- 8.1 years, BMI was 23.5 +/- 3.9 kg/m2, dietary calcium intake was 1066.1 +/- 443.3 mg per day. Thirty-eight percent of patients were lactose intolerant. Sixty-two percent of patients were in the light weight bearing activity group. The mean +/- SD preoperative BMD was 119.2 +/- 34.9 mg/cc, with T-scores of - 1.77 +/- 1.22 and Z-scores of 0.43 +/- 1.27. A decrease in BMD during the first 6 months ( approximately 13%) was statistically significant (P = 0.0001) and continued further during next 6 months (BMD loss of approximately 18% at 12 months). Patients with osteoporosis, as defined by T scores < or = - 2.5, increased from 24% at baseline to 48% at 6 months. Nonsmokers, nonalcoholics, patients with higher physical activity, and patients with a BMI > 25 kg/m2 had statistically significant higher BMD compared with their counterparts (P < 0.05). Body weight < 60 kg and BMI < 25 kg/m2 were significant risk factors for loss of BMD (P < 0.05). Dietary calcium had a discernible but statistically insignificant effect on BMD (P = 0.16). Lactose intolerance had no significant effect on BMD or bone loss. CONCLUSIONS: Osteoporosis was common in the population affected by prostate carcinoma. Orchidectomy led to accelerated bone loss. Periodic measurement of BMD after ADT would help in the early detection of osteoporosis. Maintenance of high BMI, weight-bearing physical activity, avoidance of alcohol and smoking, and possibly high dietary calcium intake help in maintaining bone mass. PMID- 15830348 TI - Disease biology rather than age is the most important determinant of survival of patients > or = 60 years with acute myeloid leukemia treated with uniform intensive therapy. AB - BACKGROUND: The objectives of the current study were to evaluate the outcome of patients > or = 60 years with acute myeloid leukemia (AML) treated uniformly with high-dose daunorubicin containing induction and modified high-dose cytosine arabinoside containing postremission therapy, and to identify factors predictive of complete disease remission (CR) and survival. METHODS: Between 1998 and 2002, the authors treated 117 newly diagnosed patients (acute promyelocytic leukemia excluded) with AML > or = 60 years (median, 67 years; range, 60-82 years). Karyotype (Medical Research Council classification) at diagnosis was categorized as good risk (n = 3), intermediate risk (n = 69), adverse risk (n = 26), and suboptimal/not done (n = 19). A normal karyotype was seen in 41 patients and 40 (34%) had secondary AML. RESULTS: The outcome of induction included the following: CR, 62 (53%); early death, 5 (4%); death during hypoplasia, 14 (12%); and resistant disease, 36 (31%). The 3-year event-free (EFS) and overall survival (OS) rates were 9% (95% confidence interval [95% CI], 3-16%) and 17% (95% CI, 9 29%), respectively. In a univariate analysis, cytogenetics, lactate dehydrogenase level, leukocyte count, and performance status were the significant factors for EFS and OS. Age was not a significant prognostic factor for either CR or survival. In a multivariate model, adverse-risk cytogenetics, previous history of myelodysplastic syndrome or antecedent hematologic disorder, and high leukocyte count (> 30 x 10(9)/L) were independent adverse prognostic factors for survival. The impact of adverse karyotype on EFS and OS was time dependent and was observed after 50 and 150 days, respectively. CONCLUSIONS: The authors concluded that candidacy for intensive therapy in older patients should be based on biologic features of disease and fitness, rather than on age. PMID- 15830349 TI - Region-specific expression of mario reveals pivotal function of the anterior nondigit region on digit formation in chick wing bud. AB - We report the region-specific expression of a novel gene, named mario, whose expression domain is in the distal tip of the presumptive and developing digit 2 region in the developing chick wing bud. The anterior region-specific expression of mario corresponds well with the presence of digit 2, and fate map analysis showed that mario expression at early stages represents the presumptive digit 2 region. Using mario expression as a region-specific marker for the digit 2 region, several surgical operations were performed to obtain insights into digit 2 development in the chick wing. Cell fate tracing concomitant with a zone of polarizing activity (ZPA) implantation revealed that an additional digit 2 in the ZPA implantation into the anterior or middle region of wing bud is derived from the original digit 2 region (mario-positive region). Surgical manipulations revealed that the anterior nondigit region has an inhibitory effect on digit 2 formation. Taken together, these results suggest that the most-anterior region, including the anterior necrotic zone, restricts the position of digit 2 region by limiting the anterior border of the digit 2 region and preventing its expansion. PMID- 15830350 TI - Prognostic value of CD44 isoform expression in thymic epithelial neoplasms. AB - BACKGROUND: Many histologic classifications of thymic epithelial tumors have been reported to date, but to the authors' knowledge, none of them closely reflect the clinical behavior or prognosis of the tumor. Therefore, it is necessary to establish a biologic marker for thymic epithelial tumors. Variants of CD44 may be important in promoting tumor progression and metastasis. Accordingly, the expression of CD44 isoforms in thymic epithelial neoplasms was investigated using immunohistochemistry to assess their possible value as prognostic indicators. METHODS: Expression of CD44v6 in thymic epithelial tumors was investigated with immunohistochemistry using consecutive surgical specimens resected from 108 patients between 1983 and 2002 at Juntendo University Hospital in Tokyo, Japan. RESULTS: Among the 108 thymic epithelial tumors, 70 were negative for CD44v6, 20 were weakly positive, and 18 were strongly positive. The status of CD44v6 expression (negative vs. weakly plus strongly positive) was found to be correlated with the tumor stage according to the Masaoka staging system (noninvasive vs. invasive tumors) (P = 0.0214). When patients with tumors that were negative and weakly positive for CD44v6 expression were combined, the 5 year, 10-year, and 15-year recurrence-free survival rates were 98.2%, 95.9%, and 86.1%, respectively, whereas the corresponding rates for patients with strongly positive tumors were 73.5%, 73.5%, and 55.1%, respectively. Therefore, these two groups demonstrated a significant difference with regard to recurrence-free survival (P = 0.0172). CONCLUSIONS: CD44v6 expression in thymic epithelial neoplasms demonstrated a significant difference based on the World Health Organization classification, the Masaoka stage (invasive vs. noninvasive tumors), and recurrence, if an appropriate cutoff value was chosen in each case. This suggests that CD44v6 can be used as a marker that reflects the stage of thymic tumors. PMID- 15830351 TI - Aromatase inhibition reduces expression of FOXL2 in the embryonic chicken ovary. AB - P450-aromatase is the terminal estrogen-synthesizing enzyme and a key gene in avian sex determination. Aromatase is expressed specifically in female gonads, but not male gonads, at the onset of sexual differentiation. This enzyme shows temporal and spatial colocalization with the forkhead transcription factor FOXL2 in the embryonic chicken ovary, suggesting a causal link. Mutations in FOXL2 are associated with premature ovarian failure in humans. Foxl2 null mice also present with premature ovarian failure. Here, we show that FOXL2 expression is reduced but not abolished in chicken embryos subjected to experimental female to male sex reversal with an aromatase inhibitor. This finding suggests that FOXL2 lies upstream of aromatase in avian sex determination, but that it responds to depleted estrogen synthesis. The reduction in FOXL2 expression may be accounted for by interruption of a positive feedback loop by means of estrogen, or the influence of testis promoting factors such as SOX9 and DMRT1 in the masculinized gonads. PMID- 15830352 TI - Dynamic balance between activation and repression regulates pre-mRNA alternative splicing during heart development. AB - Cardiac troponin T (cTNT) exon 5 splicing is developmentally regulated such that it is included in embryonic but not adult heart. CUG-BP and ETR-3-like factor (CELF) proteins promote exon inclusion, whereas polypyrimidine tract binding protein (PTB) and muscleblind-like (MBNL) proteins repress inclusion. In this study, we addressed what happens to these regulatory proteins during heart development to shift the regulatory balance of cTNT alternative splicing. Using dominant-negative proteins, we found that both CELF and PTB activities are required for appropriate splicing in cardiomyocytes. Two CELF proteins, CUG-BP and ETR-3, are nuclear and cytoplasmic in embryonic heart but are down-regulated in adult heart concomitant with loss of exon inclusion. In contrast, PTB and MBNL1 are expressed throughout heart development. The patterns of cTNT splicing and expression of its regulatory factors are conserved between mouse and chicken. Thus, alternative splicing is determined by a balance between positive and negative regulation, and modulation of expression levels of auxiliary splicing regulators may drive developmental splicing changes. ETR-3 and CUG-BP proteins are also down-regulated in other tissues during development, suggesting that CELF proteins play a broad role in developmental splicing regulation. PMID- 15830353 TI - Expression of Fgf receptors 1, 2, and 3 in the developing mid- and hindbrain of the mouse. AB - Fibroblast growth factor 8 (FGF8) mediates the function of the midbrain-hindbrain organizer (MHO). FGF signals are transmitted by means of four known FGF receptors (FGFRs). Studies of Fgfr expression in early vertebrate development have shown that Fgfr1 is expressed along the entire neural tube, whereas Fgfr2 and Fgfr3 expression has been shown to spare the tissue adjacent to the MHO. The FGF8 signal from the MHO, therefore, was believed to be transmitted by FGFR1 exclusively. However, incongruent results from conditional mutants of Fgf8 and Fgfr1 in the midbrain-hindbrain (MHB) region contradict this hypothesis. Therefore, we reexamined the expression of the Fgfrs in this region. Fgfr1 is expressed all over the neural tube. Strikingly, Fgfr2 is expressed throughout the floor plate of the MHB region. In the basal plate, Fgfr2 directly abuts the Fgf8 expression domain at the MHO, anteriorly and posteriorly. Fgfr3 expression is in contact with the Fgf8 expression domain only in the rostroventral hindbrain. Based on these findings, we postulate a role for FGFR2 and FGFR3 in FGF signaling in the ventral midbrain and hindbrain. PMID- 15830355 TI - Boundary formation in the Drosophila wing: functional dissection of Capricious and Tartan. AB - Cells in multicellular organisms often do not intermingle freely with each other. Differential cell affinities contribute to organizing cells into different tissues. Drosophila limbs and rhombomeres of the vertebrate hindbrain are subdivided into compartments. Cells in adjacent compartments do not mix. The wing primordium is subdivided into dorsal (D) and ventral (V) compartments by the activity of the LIM-homeodomain protein Apterous in D cells. The leucine-rich repeats transmembrane proteins Capricious and Tartan have been shown to contribute to formation of the affinity boundary between dorsal and ventral compartments. Here, we have carried out a structure-function analysis of Capricious and Tartan. We present evidence that both the extracellular and intracellular domains are required for the establishment of a DV affinity boundary. Our data suggest that the extracellular domains of Capricious and Tartan may work as ligands of an unknown D cell surface protein. Their intracellular domains may be required to transduce a signal necessary for the establishment of the DV boundary. PMID- 15830356 TI - Noncorrelating Pap tests and cervical biopsies: histological predictors of subsequent correlation. AB - Lack of correlation between dysplastic cervicovaginal Papanicolaou (Pap) tests and subsequent cervical biopsies raises the concern that a significant squamous intraepithelial lesion (SIL) may go unconfirmed. Additional tissue sections of cervical biopsies may detect SILs after noncorrelation on initial sections. Complete step sectioning of paraffin blocks was undertaken on 111 noncorrelating biopsy specimens from 95 patients and selected slides were reviewed for the presence of SIL. The initial negative biopsy slides were evaluated for four histological features: chronic cervicitis, acute cervicitis, mucosal erosion, and squamous atypia. Twenty-seven biopsies (24.3%) demonstrated the presence of a SIL in deeper levels. The presence of squamous atypia was significantly associated with the presence of dysplasia deeper in the block (P < 0.002). Acute and chronic cervicitis was seen roughly equally. Additional tissue levels are a productive way of confirming SILs, and squamous atypia allows a refined selection of negative cervical biopsies most likely to reveal an SIL on review of deeper levels. PMID- 15830357 TI - Beware of chasing crystals in sputum samples. PMID- 15830358 TI - A case of uterine adenomyoma with bizarre smooth muscle cells mimicking leiomyosarcoma. AB - We report a case of adenomyoma of the uterus that was cytologically difficult to distinguish from leiomyosarcoma. Examination of a uterine cervical smear revealed numerous spindle cells that were present in cell clusters or as isolated cells. These cells contained nuclei that were oval-shaped/elongated with nucleoli and delicate wispy cytoplasm. Large and bizarre nuclei were also identified. Based on these cytological findings leiomyosarcoma was considered: however, this diagnosis remained uncertain because of the absence of mitosis and/or necrotic substance. Histologically, we recognized leiomyomatous smooth muscle cells growing in a solid pattern and intermingled with endometrial-type glands. Moreover, bizarre smooth muscle cells were observed in the surface layer of the tumor. These observations suggest that for a diagnosis of uterine leiomyosarcoma the presence of mitosis and/or necrosis is important in addition to nuclear atypia. PMID- 15830359 TI - Rhabdomyosarcoma in an adult presenting with nodal metastasis: a pitfall in fine needle aspiration cytology of lymph nodes. AB - A 40-yr-old woman with an asymptomatic sinonasal rhabdomyosarcoma (RMS) initially presented with submental nodal metastasis. The fine-needle aspiration (FNA) and the subsequent biopsy of the nodal metastasis were misinterpreted as metastatic carcinoma because the primary tumor was occult, the tumor cells were exclusively round cells with a nested arrangement, and rhabdomyoblasts were absent. The correct diagnosis of metastatic RMS became apparent when the primary sinonasal tumor, detected in a CT, was biopsy proven to be an alveolar RMS. Retrospectively, there were helpful clues to the correct diagnosis in the nodal FNA and biopsy. When FNA cytology or biopsy histology of a lymph node suggests metastatic carcinoma but the tumor cells are nonimmunoreactive to carcinoma markers, the differential diagnosis should be expanded to include not only metastatic melanoma but also metastatic sarcoma and lymphoma. Cytologically, the presence of multinucleated giant tumor cells, including the rosette forms, in the FNA smears should alert the cytopathologist to the possibilities of sarcoma and anaplastic large cell lymphoma. PMID- 15830360 TI - Cytopathology of uncommon malignant renal neoplasms in the pediatric age group. AB - Malignant renal neoplasms are common solid tumors in pediatric oncology practice. These include the common Wilms' tumor/nephroblastoma and the uncommon neoplasms such as clear-cell sarcoma of the kidney (CCSK), rhabdoid tumor, renal-cell carcinoma, and others. The aim of this study was to describe in detail the cytopathological features of the histopathologically proven uncommon pediatric renal tumors. Aspirates from Wilms' tumor, which are mesenchyme predominant, show clusters of spindle cells associated with the matrix material. Evidence of rhabdomyoblastic differentiation may be present. CCSK, classic subtype, is characterized by round to oval cells arranged perivascularly and also in sheets and clusters intimately associated with a metachromatic matrix mucopolysaccharide material better appreciated in May-Grunwald-Giemsa (MGG)-stained smears. The cells also have more abundant cytoplasm and may show nuclear grooves. Spindle cell pattern of CCSK is difficult to diagnose on aspiration cytology. Renal-cell carcinoma of childhood shows similar cytological features as its adult counterpart. Rhabdoid tumor of the kidney is characterized by a monomorphic population of cells with abundant cytoplasm, eccentric nuclei with prominent nucleoli. Intrarenal yolk sac tumor is a rare neoplasm and shows severely pleomorphic cells on aspiration. Awareness of these entities is important for the practicing cytopathologist. Further, non-Wilms' renal malignant neoplasms must be distinguished from the common Wilms' tumor so that appropriate chemotherapy protocols may be instituted in cases where the tumor is in an advanced stage of malignancy. PMID- 15830361 TI - Natural killer cells in cerebrospinal fluid of a 31/2-month-old infant with DiGeorge syndrome and CMV infection. PMID- 15830362 TI - Intranuclear cytoplasmic inclusions in fine-needle aspiration smears of papillary thyroid carcinoma: a study of its morphological forms, association with nuclear grooves, and mode of formation. AB - Intranuclear cytoplasmic inclusion (INCI) and nuclear grooves in fine-needle aspiration (FNA) smears of papillary thyroid carcinoma (PTC) represent cytoplasmic invaginations into the nucleus. Although formation of INCIs is linked to nuclear grooves, they seldom exist together. This study was undertaken to find out the various morphological forms of INCIs, their relation to nuclear grooves, and mode of formation. FNA smears of 54 PTC cases were studied for various forms of INCI, nuclear chromatin pattern, nuclear grooves, and nuclear notches. A differential count of INCIs was made in 19 cases having > or =10 INCIs per 200 neoplastic cells. INCIs were present in 48 (88.9%) of 54 PTC cases. Round INCIs were present in 46 (85.2%) cases, followed by oval (48.1%), tear drop/flask shaped (18.5%), irregular (14.8%), planoconvex/semicircular (13%), rectangular (9.3%), spindle-shaped (3.7%), and bilobed (3.7%). A differential count of INCIs also showed that the round form was the commonest (76.3%). The oval and other forms constituted 19.5 and 4.2%, respectively. INCIs and nuclear grooves were present together in a cell in 15 (27.8%) cases. The formation of INCIs as a cytoplasmic invagination into the nucleus was shown cytomorphologically in rare cells. For the first time in this study, the various morphological forms of INCIs, and the extent of their coexistence with nuclear grooves, (have been highlighted) and showed their mode of formation shown at a light microscopic level. PMID- 15830363 TI - Fine-needle aspiration of primary osteosarcoma: a cytological-histological study. AB - This study aims to determine the diagnostic accuracy of fine-needle aspiration (FNA) evaluation of primary osteosarcoma (OS) and to review diagnostic criteria and adjunctive methods that can contribute to a confident diagnosis of OS in cytological smears. We evaluated FNA smears of OS in 59 patients for the following: cytomorphology and occurrence of osteoid, usefulness of adjunctive methods in evaluation of FNA smears and correspondence of FNA to the clinical data, and the histological features of excised tumors. Reliable cytological criteria of malignancy were found in 49 smears of high-grade OS. These criteria, correlated with radiographic studies and complemented by ancillary techniques, allowed diagnosis of OS or suspicion of OS in 44 cases. An additional four smears were diagnosed as sarcoma and one case was diagnosed erroneously as being carcinoma metastasis. There were no false positive or false negative diagnoses. We conclude that FNA smears from high-grade OS have characteristic features, which together with clinical and radiological data and ancillary studies allow correct diagnosis in most tumors. PMID- 15830365 TI - The diagnosis of trichomonas vaginalis in liquid-based Pap tests: morphological characteristics. AB - We investigated the morphological features of Trichomonas vaginalis in liquid based Papanicolaou (Pap) (LBP) tests, in order to assess the impact of this new slide preparation system on recognition of T. vaginalis organisms. We reviewed 88 LBP test slides previously interpreted as showing T. vaginalis and assessed morphological characteristics of the organisms and the presence of secondary features. Eighty-six of 88 slides showed, at least focally, organisms with cytoplasmic granules, nuclei, and flagella allowing for confident recognition of T. vaginalis. Compared with 58 controls, cases of T. vaginalis were more likely to have squamous cells with perinuclear halos, "ghost cells," and coccobacilli. The lack of a "dirty" background or cytolysis allowed a more straightforward identification of the organisms. We concluded that T. vaginalis organisms are morphologically distinct in LBP tests, which may lead to an increased specificity when they are identified in such tests. PMID- 15830366 TI - Primary ovarian angiosarcoma presenting as malignant cells in ascites: case report and review of the literature. AB - Primary angiosarcoma of the ovary is a rare tumor, with less than 25 cases reported in the literature. These tumors are thought to arise from carcinosarcomas, teratomas or the ovarian vasculature, and occur at any age. The prognosis is dismal in the majority of cases. We present the case of a 19-year old female who presented with abdominal distention and abdominal pain. Tapping of peritoneal effusion showed groups of pleomorphic cells that were negative for epithelial and germ cell markers. Immunohistochemistry performed on the surgical specimen of the ovary, and subsequently on the effusion specimen showed staining for endothelial markers. The patient was treated post-operatively with chemotherapy, but died one year following diagnosis. This is the first reported case of an ovarian angiosarcoma that metastasized to the peritoneal cavity, with a resulting malignant effusion. Despite the rarity of metastasis from gynecological sarcomas in effusions, this possibility needs to be included in the differential diagnosis of malignant effusions that are negative for epithelial and germ cell markers. PMID- 15830367 TI - The continuing role of fine-needle aspiration of the prostate gland into the 21st century: a tribute to Torsten Lowhagen. AB - A brief review of the history of transrectal fine-needle aspiration (FNA) of the prostate gland is reported in this article; the authors'experience of FNA during the last 20 yr is described also. Despite the worldwide acceptance of the thin needle core approach, the use of transrectal FNA of palpable abnormalities of the prostate still is advocated because it is cheaper, faster, easier to perform, and results in lower morbidity than any other technique so far developed. High sensitivity, specificity, and efficacy account for its reliability. Appropriate training in performing transrectal FNA of the prostate and in interpreting the smears is, of course, essential. Transrectal FNA should be the initial diagnostic procedure for suspected prostatic cancer and will continue to be a useful diagnostic tool in the 21st century. PMID- 15830368 TI - Cervical chordoma masquerading as a thyroid neoplasm: a case report. AB - Chordomas are rare neoplasms of notochord derivation that arise along the craniospinal axis. Their incidence along the cervical spine and close proximity to the thyroid can pose diagnostic pitfalls. We report a case of a paraspinal mass juxtaposed adjacent to the thyroid with intranuclear inclusions of papillary carcinoma, abundant eosinophilic cytoplasm of an oncocytic lesion, and pleomorphism of anaplastic carcinoma that expressed thyroglobulin and emulated a malignant thyroid neoplasm. However, subsequent histological examination revealed a chordoma with perithyroidal soft-tissue invasion. This case highlights the significance of including chordoma in the differential diagnosis of a thyroid neoplasm and interpreting thyroglobulin reactivity warily, because its inadvertent tracking during fine-needle aspirates (FNAs) can produce false positive staining of nonthyroidal cells. PMID- 15830369 TI - FNA diagnosis of dermatofibrosarcoma protuberans. AB - Fine-needle aspiration (FNA) features of primary and recurrent dermatofibrosarcoma protuberans (DFSP) in two patients were evaluated and correlated to ancillary studies and clinical data. The tumors developed in the skin and subcutaneous tissue of the back in both patients. The cytological findings included clusters and dispersed uniform or slightly atypical spindle cells, collagenous matrix, and, in one case, cell clusters with a storiform pattern. Both tumors were diagnosed correctly when FNA was complemented by ancillary studies (immunostaining) and clinical data. FNA features alone are not sufficiently characteristic to permit a confident diagnosis of DFSP. Correct diagnosis requires an awareness of the clinical history and the use of ancillary studies. PMID- 15830370 TI - Fine needle aspiration cytology of lobular carcinoma in situ on ThinPrep. AB - Lobular carcinoma in situ (LCIS) of the breast is a recognized marker for increased risk of invasive carcinoma and has well-established histologic criteria. However, its detection and diagnosis on FNA of breast has not been well defined. Cytology slides (all ThinPrep) of 11 cases with biopsy-proven LCIS at Beth Israel Deaconess Medical Center were reviewed. All 11 cases showed tight and/or loosely cohesive clusters of crowded mildly enlarged nuclei and ten of 11 showed at least moderate cellularity. Single epithelial cells, small but prominent nucleoli, intracytoplasmic lumina, and two distinct epithelial-cell populations were also noted in some cases. As none of these features is specific for LCIS, it would be prudent to report such lesions as atypical so that a core biopsy or excisional biopsy will be performed before definitive treatment. The original FNA diagnosis of the 11 cases ranged from epithelial proliferation without atypia to carcinoma. Three of the 11 cases had fibroadenomas on histology with extensive involvement by LCIS. Since management for LCIS is different from that for invasive carcinoma or DCIS, it should be considered and distinguished from the latter two in cases suspicious for carcinoma on FNA. PMID- 15830371 TI - FNA diagnosis of primary adult onset lymphocutaneous Langerhans' cell histiocytosis masquerading as deep fungal mycosis. AB - Langerhans' cell histiocytosis (LCH) in its aggressive disseminated form seen most often in children is easily diagnosed by the treating physician. On the contrary, LCH in an adult is localized, extremely rare, and, hence, its diagnosis is missed quite often or underdiagnosed. We describe the troubleshooters encountered in the fine-needle aspiration (FNA) diagnosis of LCH in an adult who presented for 4 years with ulceronodular lesions over the neck, both axillae, and inguinal regions since 4 years of age, which had closely mimicked deep mycosis both clinically and histopathologically. PMID- 15830372 TI - Expression pattern of glypican-4 suggests multiple roles during mouse development. AB - Glypicans are glycosylphosphatidylinositol-anchored heparan sulphate proteoglycans, which are thought to modulate signalling by growth factors, including fibroblast growth factors and Wnts. Studies in Xenopus have implicated glypicans, in particular glypican-4, in the process of convergent extension, which is essential for neural tube closure in Xenopus and mouse. Expression of glypican-4 has been reported in the mouse brain at embryonic day 10 and later stages, whereas expression during the developmental stages encompassing initiation and progression of neural tube closure has not been reported. Analysis by in situ hybridization reveals a complex pattern of glypican-4 mRNA localization at embryonic days 7-10.5, including sites in the anterior forebrain neuroepithelium, branchial arches, optic and otic vesicles, limb buds and somites. Glypican-4 expression is not detected in the midline of the embryo at the stage of initiation of neural tube closure, suggesting that glypican-4 is unlikely to play an essential role in convergent extension in the mouse. PMID- 15830373 TI - Stability of PreservCyt for Hybrid Capture (HC II) HPV test. AB - The Food and Drug Administration (FDA) has approved the Hybrid Capture II (HC II) assay to test for the presence of high-risk types of human papilloma virus (HPV) DNA using specimens in PreservCyt fixative for up to 21 days after collection. The ability of HC II to determine the presence of HPV DNA in actual patient samples after longer periods of storage has not been shown. To determine if specimens older than 21 days can yield useful results, 207 patient specimens that had been tested for HPV DNA by HC II (primary test) were tested again after a significant period of storage ranging from approximately 2.5 to 13.5 mo (retest). The results of the primary test and the retest agreed in 86% of the cases. The high level of agreement in the results suggests that the presence of high-risk types of HPV DNA can be determined from actual cervical cytology material in PreservCyt with the HC II assay for at least 3 mo after specimen collection. PMID- 15830374 TI - Myosin heavy chain expression in zebrafish and slow muscle composition. AB - In the zebrafish embryo, two distinct classes of muscle fibers have been described in the forming myotome that arise from topographically separable precursor populations. Based entirely on cross-reactivity with antibodies raised against mammalian and chick myosin heavy chain isoforms slow twitch muscle has been shown to arise exclusively from "adaxial" myoblasts, which migrate from their origin flanking the notochord to form a single layer of subcutaneous differentiated muscle cells. The remainder of the myotome differentiates behind this migration as muscle fibers recognized by anti-fast myosin heavy chain (MyHC) antibodies. To identify unambiguous molecular markers of cell fate in the myotome, we have characterized genes encoding zebrafish fast and slow MyHC. Using phylogenetic and expression analysis, we demonstrate that these genes are definitive molecular markers of slow and fast twitch fates. We also demonstrate that zebrafish embryonic slow twitch muscle co-expresses both slow and fast twitch MyHC isoforms, a property that they share with primary fibers of the amniote myotome. PMID- 15830375 TI - Mitogen-activated protein kinase expression and activation does not differentiate benign from malignant mesothelial cells. AB - BACKGROUND: In vitro studies of malignant mesothelioma (MM) cells have suggested activation of mitogen-activated protein kinase (MAPK) in response to asbestos exposure. The objective of this study was to investigate protein expression (level) and phosphorylation status (activity) of the extracellular-regulated kinase (ERK), the c-Jun amino-terminal kinase (JNK), and the high-osmolarity glycerol response kinase (p38) in vivo through the analysis of fresh frozen reactive mesothelium (RM) and MM specimens. METHODS: MAPK levels were analyzed in 36 fresh-frozen MM specimens (32 effusions, 4 biopsies) and in 14 RM specimens (all effusions) using immunoblotting with antibodies detecting the total (pan-) and activated (phospho-) fraction (p-) of ERK, JNK, and p38. Values for pan-MAPK and p-MAPK expression and the p-MAPK/pan-MAPK ratio in MM and RM specimens were compared. Results were corroborated using immunocytochemistry for p-ERK, p-JNK, and p-38 in selected specimens. RESULTS: Pan-ERK, pan-JNK, and pan-p38 expression was found frequently in both MM specimens (35 of 36 specimens) and RM specimens (14 of 14 specimens) using immunoblotting, with comparable findings for activated p-p38 (34 of 36 MM specimens, 13 of 14 RM specimens). Activation of p-ERK (27 of 36 MM specimens, 10 of 14 RM specimens) and p-JNK (25 of 36 MM specimens, 10 of 14 RM specimens) was less frequent. Pan-ERK (P = 0.016), pan-JNK (P = 0.004), pan p38 (P = 0.012), and p-ERK (P = 0.02) expression levels were higher in MM specimens from female patients. Pan-p38 expression levels also were higher in peritoneal MM specimens (P = 0.019). MM and RM showed similar MAPK expression, activation, and activation ratios (Mann-Whitney test; P > 0.05). Immunocytochemistry localized MAPK to MM and RM cells. CONCLUSIONS: The current results provided the first evidence of in vivo activation of MAPK in clinical MM and RM. The similar values in these two cell types suggest that MAPK may not be involved in the transformation of benign to malignant mesothelium, thus bringing into question the validity of using MAPKs as molecular therapeutic targets in patients with MM. PMID- 15830376 TI - Establishment of a new embryonic stem cell line derived from C57BL/6 mouse expressing EGFP ubiquitously. AB - Transgenic mice ubiquitously expressing enhanced green fluorescent protein (EGFP) are useful as marker lines in chimera experiments. We established a new embryonic stem (ES) cell line (named B6G-2) from a C57BL/6 blastocyst showing ubiquitous EGFP expression. Undifferentiated B6G-2 cells showed strong green fluorescence and mRNAs of pluripotent marker genes. B6G-2 cells were transferred into a C57BL/6 blastocyst to generate a germline chimera, the progeny of which inherited ubiquitous EGFP expression. Mice derived completely from B6G-2 cells were also developed from the ES cells; these were tetraploid chimeras. The established B6G 2 cells were shown to be pluripotent and to be capable of differentiating into cells of all lineages. Thus, the new ES cell line expressing EGFP ubiquitously is useful for basic research in the field of regenerative medicine. The B6G-2 cell line is freely available from the BioResource Center, RIKEN Tsukuba Institute (http://www.brc.riken.jp/lab/cell/english/). PMID- 15830377 TI - Planar polarity of hair cells in the chick inner ear is correlated with polarized distribution of c-flamingo-1 protein. AB - Hair cells of the vertebrate inner ear are directional mechanosensors: they have a polarity, defined by a vector in the plane of the sensory epithelium. It has been suggested that this polarity might be controlled by genes homologous to those that control planar cell polarity (PCP) in Drosophila, and vertebrate homologues of the Drosophila PCP genes Van Gogh/strabismus and flamingo/starry night are indeed essential for normal hair cell PCP. The underlying molecular mechanism is unclear, however. Although the PCP protein Flamingo shows a polarized intracellular distribution in the fly, it is unknown whether this is necessary for its function. Here, we describe the expression pattern of a flamingo homologue, c-flamingo-1 (c-fmi-1), in the developing chick ear and show that its protein product, like that of flamingo in the fly, has a polarized distribution in each hair cell, defining an axis that corresponds to the structural PCP axis. This conservation between fly and vertebrate suggests that the polarized protein localization is functionally important. In the basilar papilla, the same localization is seen in supporting cells also, suggesting that supporting cells are cryptically polarized, despite having no overt structural polarity; they may thus participate in PCP signal transmission across the sensory patch. PMID- 15830378 TI - Design and characterization of a compact dual channel virus counter. AB - BACKGROUND: Although there is a growing need in the field of biotechnology to rapidly and accurately quantify viruses, time-consuming techniques such as the plaque titer method remain the "gold standard." Flow cytometric methods for virus quantification offer the advantages of rapid analysis and statistical treatment. The technique presented in this work represents the first demonstration of a flow cytometric determination of a viral count that is directly related to the count obtained by plaque titer. METHODS: A flow cytometric instrument for rapid quantification of virus particles was designed, constructed, and thoroughly characterized. A two-color method, which involved staining the viral genome and the protein coat for baculoviruses, was developed in addition to an algorithm to identify simultaneous events on the DNA and protein channels. RESULTS: The instrument was fully characterized, which included analysis of the data acquisition rate, sampling time, flow rate, detection efficiency, linear dynamic range, channel cross-talk, and the limit of detection. Baculovirus samples were analyzed and the results were compared with concentrations obtained by a one channel flow cytometer and plaque assay. CONCLUSIONS: The dual channel virus counter yields a representative value for the concentration of active viruses in an unpurified sample when compared with plaque assay and a one-channel flow cytometer. The technique is rapid (within minutes), requires only minimal sample preparation and minimum sample size (approximately 100 microl). PMID- 15830379 TI - DARPP-32 genomic fragments drive Cre expression in postnatal striatum. AB - To direct Cre-mediated recombination to differentiated medium-size spiny neurons (MSNs) of the striatum, we generated transgenic mice that express Cre recombinase under the regulation of DARPP-32 genomic fragments. In this reported line, recombination of an R26R reporter allele occurred postnatally in the majority of medium-size spiny neurons of the dorsal and ventral striatum (caudate nucleus and nucleus accumbens/olfactory tubercle), as well as in the piriform cortex and choroid plexus. Although regulatory fragments were selected to target MSNs, low levels of Cre-recombinase expression, as detected by beta-galactosidase activity from the R26R reporter gene, were also apparent in widely dispersed areas or cells of the forebrain and hindbrain. These included the primary and secondary motor cortex, and association cortex, as well as in the olfactory bulb and cerebellar Purkinje cells. Notably, expression in these regions was well below that of endogenous DARPP-32. Analysis of colocalization of beta-galactosidase, as detected either by histochemistry or immunocytochemistry, and DARPP-32 revealed double-labeling in almost all DARPP-32-expressing MSNs in the postnatal striatum, but not in extrastriatal regions. The DARPP-32Cre transgenic mouse line thus provides a useful tool to specifically express and/or inactivate genes in mature MSNs of the striatum. PMID- 15830380 TI - Polychromatic (eight-color) slide-based cytometry for the phenotyping of leukocyte, NK, and NKT subsets. AB - BACKGROUND: Natural killer (NK) and NK T (NKT) cells are important in innate immune defense. Their unequivocal identification requires at least four antigens. Based on the expression of additional antigens, they can be further divided into functional subsets. For more accurate immunophenotyping and to describe multiple expression patterns of leukocyte subsets, an increased number of measurable colors is necessary. To take advantage of the technologic features offered by slide-based cytometry, repeated analysis was combined with sequential optical filter changing. METHODS: Human peripheral blood leukocytes from healthy adult volunteers were labeled with antibodies by direct or indirect staining. Tandem dyes of Cy7 (phycoerythrin [PE]-/allophycocyanin [APC]-Cy7), Cy5.5 (PE-/APC Cy5.5), and PE-Cy5 and the fluorochromes fluorescein isothiocyanate (FITC), PE, and APC were tested alone and in combinations. Optical filters of the laser scanning cytometer were 555 DRLP/BP 530/30 nm for photomultiplier tube (PMT) 1/FITC, 605 DRLP/BP 580/30 nm for PMT 2/PE, 740 DCXR/BP 670/20 nm for PMT 3/Cy5/APC, and BP 810/90 nm for PMT 4/Cy7. Filter PMT 3 was replaced for detection of PE/Cy5.5 and APC/Cy5.5 by 740 LP/BP 710/20 nm and the sample was remeasured. Both data files were merged into one to combine the different information on a single-cell basis. The combination of eight antibodies against CD3, CD4, CD8, CD14, CD16, CD19, CD45, and CD56 was used to characterize NK and NKT cells and their subsets. RESULTS: In this way Cy5.5 is measurable at 488-nm and 633-nm excitation. Further, with the two different filters it is possible to distinguish Cy5 from Cy5.5 in the same detection channel (PMT 3). With this method we identified NK and NKT cells, subsets of NK (CD3-16+56+, CD3-16+56-, CD3 16-56+) and NKT (CD3+16+56+, CD3+16-56+) and their CD4+8-, CD4-8+, CD4-8- and CD4+8+ subsets. CONCLUSION: With our adaptations it is possible to discriminate tandem conjugates of Cy5, Cy5.5, and Cy7 for eight-color immunophenotyping. Using this method, novel rare subsets of NK and NKT cells that are CD4/CD8 double positive are reported for the first time. PMID- 15830381 TI - Expression and regulation of mouse SERDIN1, a highly conserved cardiac-specific leucine-rich repeat protein. AB - Despite recent progress, the precise mechanisms responsible for vertebrate cardiac development are still enigmatic. Better understanding of cardiac biology and disease necessitates identification and analysis of a full spectrum of regulatory and structural proteins specific to the developing heart. By performing an in silico screen, we identified a cardiac-specific gene we named Serdin1. The Serdin1 gene is conserved, and the message is restricted to the heart in several vertebrate species, thus implicating Serdin1 as an important gene in cardiac development. In situ hybridization confirmed that the Serdin1 message is cardiac-specific in mice as early as embryonic day 8.5. Antibody staining demonstrated predominantly nuclear staining in immortalized cardiac cell lines (P19 and HL-1) and proliferating cultured cardiomyocytes, whereas in vivo SERDIN1 localizes to I bands of the sarcomere. Seven kilobases of the upstream regulatory sequence of Serdin1 is sufficient for cardiac-specific expression. Computer analysis revealed an 80-bp homologous region between the mouse and the human Serdin genes that contains GATA, SRF, and MEF sites. Cardiac specificity and localization patterns suggest that SERDIN1 is intimately integrated with the molecular pathways controlling cardiogenesis in vertebrates. PMID- 15830382 TI - Mechanics and function in heart morphogenesis. AB - For years, biomechanical engineers have studied the physical forces involved in morphogenesis of the heart. In a parallel stream of research, molecular and developmental biologists have sought to identify the molecular pathways responsible for embryonic heart development. Recently, several studies have shown that these two avenues of research should be integrated to explain how genes expressed in the heart regulate early heart function and affect physical morphogenetic steps, as well as to conversely show how early heart function affects the expression of genes required for morphogenesis. This review combines the perspectives of biomechanical engineering and developmental biology to lay out an integrated view of the role of mechanical forces in heart development. PMID- 15830383 TI - Transgenic mice that express Cre recombinase in hypertrophic chondrocytes. AB - In order to investigate the physiological control of hypertrophic chondrocytes which present the terminally differentiated form of chondrocytes, we generated a mouse line expressing the Cre recombinase under the control of the mouse type X collagen (Col10a1) promoter. In situ hybridization analysis demonstrated the expression of Col10a1-Cre transgene in hypertrophic chondrocytes of femur at postnatal day 2 (P2). In order to test the excision activity of the Cre recombinase, the Col10a1-Cre transgenic line was crossed with the mouse strain carrying the Smad4 conditional alleles (Smad4co/co) and the reporter line ROSA26. Multiple tissue PCR of Col10a1-Cre;Smad4co/+ mice revealed the restricted Cre activity in tissues containing hypertrophic chondrocytes. LacZ staining revealed that the Cre activity was observed in the cartilage primordia of ribs at E14.5 and only detected in the lower hypertrophic region of ribs at P1. These data suggest that the Col10a1-Cre mouse line described here could be used to achieve conditional gene targeting in hypertrophic chondrocytes. PMID- 15830385 TI - Expression analysis of jagged genes in zebrafish embryos. AB - The interaction of transmembrane Delta and Jagged/Serrate ligands with Notch receptors on neighboring cells is critically involved in cell specification during development. In zebrafish, the early expression of delta but not of jagged genes has been investigated in some detail. We have analyzed the sequence and embryonic expression pattern of the three zebrafish genes jagged1a, jagged1b, and jagged2. These genes, whose transcripts are detectable by in situ hybridization from early somitogenesis, are widely and dynamically expressed in embryos. Coexpression is limited, however, to the notochord and lens (jagged1a and jagged1b) and to the otic vesicle and pronephros (jagged1b and jagged2). Conversely, jagged1a and jagged2, both widely expressed in the central nervous system, are not coexpressed. jagged2 is also detected in the epidermis, newly formed somites, pharyngeal pouches, and pancreatic exocrine anlage and jagged1b in otic placodes and cell clusters close to the pancreatic islet. The similarities of the expression patterns of jagged and delta genes in zebrafish suggest that the Jagged and Delta ligands are functionally redundant or required in specific combinations in many differentiation processes. PMID- 15830384 TI - Toll-like receptor signaling inhibits structural development of the distal fetal mouse lung. AB - We tested the hypothesis that innate immune signaling in utero could disrupt the structural development of the fetal lung, contributing to the pathogenesis of bronchopulmonary dysplasia. Injection of Escherichia coli lipopolysaccharide (LPS) into the amniotic fluid of E15 BALB/cJ mice increased the luminal volume density of fetal mouse lungs at embryonic day (E) 17 and E18. LPS also increased luminal volume and decreased distal lung branching in fetal mouse lung explants. This effect required NF-kappaB activation and functional Toll-Like Receptor 4. Airway branching may require fibronectin-dependent epithelial-mesenchymal interactions, representing a potential target for innate immune signaling. Anti fibronectin antibodies and LPS both blocked distal lung branching. By immunofluorescence, fibronectin localized to the clefts between newly formed airways but was restricted to peripheral mesenchymal cells in LPS-exposed explants. These data suggest that LPS may alter the expression pattern of mesenchymal fibronectin, potentially disrupting epithelial-mesenchymal interactions and inhibiting distal airway branching and alveolarization. This mechanism may link innate immune signaling with defects in structural development of the fetal lung. PMID- 15830386 TI - Chylothorax in Henoch-Schonlein purpura: a case report and review of the literature. AB - Henoch-Schonlein purpura (HSP) is the most common acute vasculitis in the pediatric population, with an incidence of 10-14 per 100,000. The classic presentation of this disorder includes erythematous papules followed by palpable purpura in the lower extremities, trunk, and face, arthralgia or arthritis, abdominal pain, gastrointestinal bleeding, and nephritis. While renal abnormalities in HSP are common, the classic pulmonary manifestations, such as hemorrhage and pneumonitis, are thought to be infrequent. Subclinical pulmonary manifestations, including diffusion defects and radiographic anomalies, seem to be quite frequent in patients with HSP but are not commonly reported. Other respiratory manifestations include pleural effusion and chylothorax, but these are rarely mentioned in the literature. Chylothorax was only reported once in an adult patient with HSP in whom the mechanism of formation was demonstrated to be secondary to transdiaphragmatic passage of chylous fluid from the peritoneal cavity. Here we describe an 8-year-old girl with HSP, nephrotic syndrome, and chylothorax, and we report the results of a review of the literature regarding respiratory complications in HSP. The present case is the first pediatric patient reported with HSP and chylothorax. The therapeutic measures utilized were effective in resolving her edema, ascites, and chylothorax, and we advocate the use of these measures as first-line therapy in future patients with similar complications from HSP. PMID- 15830387 TI - Developmental paradigm for early features of cystic fibrosis. AB - Cystic fibrosis (CF) is a progressive disease in which the lung is perceived to be normal at birth and is injured by recurrent infection. However, there is increasing evidence that the lung is functionally and structurally abnormal prior to the appearance of clinical infection. The cystic fibrosis transmembrane regulator (CFTR) is highly expressed in fetal tissues, and this review examines the role of CFTR in regulatory cascades during lung development. Early changes in the CF lung are examined from a perspective of disrupted fetal development, explaining a number of paradoxes seen with the disease. PMID- 15830388 TI - How much do Pseudomonas biofilms contribute to symptoms of pulmonary exacerbation in cystic fibrosis? AB - Recent studies suggest that chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) involve anaerobic biofilms, and that these biofilms are the reason chronic infections are rarely eradicated by antibiotic therapy, regardless of the in vitro susceptibility of infecting bacteria. These observations led to the development of an in vitro method for testing antibiotic susceptibility of CF clinical isolates in biofilms (Moskowitz et al., J Clin Microbiol 2004;42:1915 1922) and unearthed an apparent paradox. Antibiotics that remain cornerstones of clinical management of CF pulmonary exacerbations (e.g., beta-lactam antibiotics) appear to have little to no activity at clinically achievable concentrations when tested in vitro against clinical P. aeruginosa isolates growing in biofilms. The proven clinical efficacy of beta-lactam antibiotics in treating exacerbations, and the selection for beta-lactam resistance in vivo, suggest that planktonic bacteria play a significant role in pulmonary exacerbations of CF. A model of infection architecture is proposed in which biofilm and planktonic compartments each play a role in infection persistence and pulmonary exacerbation, respectively. Infection architecture may partially account for the observed lack of correlation between in vitro antibiotic susceptibility testing and clinical response to antibiotic therapy. PMID- 15830389 TI - Skin-prick testing as a diagnostic aid for childhood asthma. AB - Diagnosing asthma is problematic when based solely on reported symptoms. The purpose of this study was to evaluate skin-prick testing as a diagnostic aid for asthma in children. Skin-prick testing (SPT) was undertaken in children aged 2-10 years with either no history of wheeze (n = 149) or recent doctor-observed wheeze which responded to treatment with a bronchodilator, the "gold standard" (n = 164). Children with moderate or severe asthma were excluded. SPT positivity increased sharply at age 5 years in wheezers. Data were therefore divided into two age groups: 2- < 5 years (57 controls, 97 wheezers) and 5-10 years (92 controls, 67 wheezers). The sensitivity, specificity, and likelihood ratios of SPT positivity for wheeze were 32%, 89%, and 2.9, respectively, in the younger children, and 82%, 85%, and 5.5, respectively, in the older children. For a prevalence of 30% for asthma, the positive predictive values of a positive SPT were 55% and 70% for the younger and older age groups, respectively. The test characteristics of SPT for helping diagnose asthma in schoolchildren are good. The prevalence of wheeze in preschool children is high, and so SPT should be helpful even in this group. We suggest that clinicians consider skin-prick testing as a diagnostic aid for asthma. PMID- 15830390 TI - Atrial natriuretic peptide in children with pneumonia. AB - Atrial natriuretic peptide (ANP) has known natriuretic, diuretic, and vasodilatatory effects. It is synthesized and stored in the atrial cells. Stretching of the atrial muscle fibers during an increase in venous return sets a response of ANP release into the blood stream. High levels of ANP were measured in a number of lung diseases. Pneumonia in children is frequently accompanied by the hyponatremia of the syndrome of inappropriate antidiuretic hormone (ADH) secretion (SIADH). High levels of ANP were found among patients with SIADH. Our objective was to determine if ANP plasma levels are altered in children with pneumonia, and to evaluate a possible correlation between severity of pneumonia and ANP levels. Blood samples from 28 children diagnosed with pneumonia were collected. Plasma ANP levels were determined by radioimmunoassay and compared to levels in 25 children without pneumonia. ANP levels in the pneumonia group (mean +/- SD, 16.02 +/- 11.69 pg/ml) increased significantly (P < 0.01) compared to levels in the control group (mean +/- SD, 7.44 +/- 9.29 pg/ml). Children in the pneumonia group also exhibited low levels of plasma sodium (mean +/- SD, 134.88 +/- 2.5 mmol/l) compared to levels in children without pneumonia (mean +/- SD, 139.77 +/- 4.15 mmol/l) (P < 0.01). There was no correlation between ANP plasma levels and severity of pneumonia. In conclusion, ANP levels in children with pneumonia, as in other lung diseases, are increased. High ANP levels may play a role in maintaining water and electrolyte equilibrium during a state of inappropriate ADH secretion accompanying pneumonia. PMID- 15830391 TI - Sweat-testing: a review of current technical requirements. AB - Sweat-testing remains a key component of the diagnostic workup for cystic fibrosis. However, it is technically challenging, especially in young children. This review covers technical aspects related to the devices and protocols associated with conducting successful tests. PMID- 15830392 TI - Lipopolysaccharide-induced tyrosine nitration and inactivation of hepatic glutamine synthetase in the rat. AB - Glutamine synthetase (GS) in the liver is restricted to a small perivenous hepatocyte population and plays an important role in the scavenging of ammonia that has escaped the periportal urea-synthesizing compartment. We examined the effect of a single intraperitoneal injection of lipopolysaccharide (LPS) in vivo on glutamine synthesis in rat liver. LPS injection induced expression of inducible nitric oxide synthase, which was maximal after 6 to 12 hours but returned toward control levels within 24 hours. Twenty-four hours after LPS injection, an approximately fivefold increase in tyrosine-nitrated proteins in liver was found, and GS protein expression was decreased by approximately 20%, whereas GS activity was lowered by 40% to 50%. GS was found to be tyrosine nitrated in response to LPS, and immunodepletion of tyrosine-nitrated proteins decreased GS protein by approximately 50% but had no effect on GS activity. Together with the finding via mass spectrometry that peroxynitrite-induced inactivation of purified GS is associated with nitration of the active site tyrosine residue, our data suggest that tyrosine nitration critically contributes to inactivation of the enzyme. In line with GS inactivation, glutamine synthesis from ammonia (0.3 mmol/L) in perfused livers from 24-hour LPS-treated rats was decreased by approximately 50%, whereas urea synthesis was not significantly affected. In conclusion, LPS impairs hepatic ammonia detoxification by both downregulation of GS and its inactivation because of tyrosine nitration. The resulting defect of perivenous scavenger cell function with regard to ammonia elimination may contribute to sepsis-induced development of hyperammonemia in patients who have cirrhosis. PMID- 15830393 TI - Epoprostenol and home mechanical ventilation for pulmonary hypertension associated with chronic lung disease. AB - Pulmonary hypertension (PH) can be associated with bronchopulmonary dysplasia (BPD) of infancy, and mortality in these pediatric patients is high without aggressive medical treatment. Continuous intravenous epoprostenol (prostacyclin) was shown to lower pulmonary artery pressures (PAP) in children with idiopathic pulmonary arterial hypertension (PAH), formerly referred to as primary pulmonary hypertension. We report on the first case of long-term home ventilation in combination with chronic intravenous epoprostenol in a child with severe pulmonary hypertension associated with chronic lung disease. This aggressive combination resulted in significant improvement in pulmonary artery pressures, substantial improvement in quality of life, and eventual discontinuation of home ventilation. PMID- 15830394 TI - A mouse model for cystic biliary dysgenesis in autosomal recessive polycystic kidney disease (ARPKD). AB - Autosomal recessive polycystic kidney disease (ARPKD) is an important cause of liver- and renal-related morbidity and mortality in childhood. Recently, PKHD1, the gene encoding the transmembrane protein polyductin, was shown to be mutated in ARPKD patients. We here describe the first mouse strain, generated by targeted mutation of Pkhd1. Due to exon skipping, Pkhd1ex40 mice express a modified Pkhd1 transcript and develop severe malformations of intrahepatic bile ducts. Cholangiocytes maintain a proliferative phenotype and continuously synthesize TGF beta1. Subsequently, mesenchymal cells within the hepatic portal tracts continue to synthesize collagen, resulting in progressive portal fibrosis and portal hypertension. Fibrosis did not involve the hepatic lobules, and we did not observe any pathological changes in morphology or function of hepatocytes. Surprisingly and in contrast to human ARPKD individuals, Pkhd1ex40 mice develop morphologically and functionally normal kidneys. In conclusion,our data indicate that subsequent to formation of the embryonic ductal plate, dysgenesis of terminally differentiated bile ducts occurs in response to the Pkhd1ex40 mutation. The role of polyductin in liver and kidney may be functionally divergent, because protein domains essential for bile duct development do not affect nephrogenesis in our mouse model. PMID- 15830395 TI - Mild chronic hepatitis C with steatosis: which treatment? PMID- 15830396 TI - Do pharmacists' opinions affect their decision to dispense or recommend a visit to a doctor? AB - PURPOSE: To assess the relation between the pharmacists' opinions and the decision to dispense drugs without medical prescription and to recommend a visit to a doctor. METHODS: We carried out a cohort study on a sample of 166 pharmacists in North-West Spain. Pharmacists' opinions on prescription practice of the doctors, on pharmacists' qualification to dispense drugs without medical prescription, on their responsibility about dispensed drugs, on clients' qualification for self-medication, and pharmacists' perception of their work were collected through a personal interview. Dispensing and the recommendation to the patient to visit their doctor were measure in the follow-up. We constructed logistic regression models. RESULTS: The response rate to the first questionnaire was 98.8% and the participation rate in the follow-up was 60%. Pharmacists who considered that doctors prescribed excessively were less likely to dispense without medical prescription (OR = 0.48) and to send the client to the doctor more often (OR = 2.33). On the other hand, those who considered themselves to be capable to dispense without prescription do so frequently (OR = 1.24). A major appreciation of the pharmacist as a health educator was associated with a higher dispensing (OR = 3.81). Pharmacists who considered that customers' qualification for self-medication was good recommended, more frequently visiting a doctor (OR = 1.58). CONCLUSIONS: Our results show that the pharmacists' opinions are associated with their practice of counseling. Therefore, any program designed with the purpose of changing dispensing habits of the pharmacists should identify and take into account their opinions. PMID- 15830397 TI - A key role for autoreactive B cells in the breakdown of T-cell tolerance to pyruvate dehydrogenase complex in the mouse. AB - The key immunological event in the pathogenesis of the autoimmune liver disease primary biliary cirrhosis is breakdown of T-cell self-tolerance to pyruvate dehydrogenase complex (PDC). The mechanism resulting in this breakdown of tolerance remains unclear. Mice exposed to self-PDC mount no immune response; however, animals coexposed to self-PDC and PDC of foreign origin (which in isolation induces a cross-reactive antibody but not an autoreactive T-cell response) show breakdown of T-cell as well as B-cell tolerance. This observation raises the possibility that a cross-reactive antibody response to self-PDC can promote breakdown of T-cell tolerance. The aim of this study was to address the hypothesis that breakdown of T-cell tolerance to PDC can be driven by the presence of B cells and/or antibodies cross-reactive with this self-antigen. Naive female SJL/J mice were exposed to self-PDC alone and in the presence of purified splenic B cells from animals primed with foreign PDC (or controls) or purified immunoglobulin (Ig) G from the same animals. Breakdown of T-cell tolerance was assessed by splenic T-cell proliferative response to antigen at 5 weeks. CD4(+) T-cell proliferative responses indicative of breakdown of T-cell tolerance to self-PDC were seen in the majority (7 of 9, 78%) of animals receiving self-PDC together with purified PDC-reactive B cells. Tolerance breakdown was not seen in animals receiving self-PDC with purified anti-PDC IgG or with B cells from animals sensitized with an irrelevant antigen. In conclusion, breakdown of T-cell tolerance to the highly conserved self-antigen PDC may be mediated by high-level presentation of self-derived epitopes by activated cross-reactive B cells. PMID- 15830398 TI - Serum paraoxonase 1 activity and malondialdehyde levels in patients with ulcerative colitis. AB - This study was designed to evaluate the oxidative and antioxidative status in patients with ulcerative colitis by detecting antioxidant enzyme paraoxonase 1 activity together with the level of a well-known marker of oxidative stress, malondialdehyde. Serum paraoxonase 1 activity and malondialdehyde levels were analysed in 30 patients with ulcerative colitis and 30 controls using a spectrophotometric method; correlation analysis was made between these variables. Serum malondialdehyde levels were higher in the ulcerative colitis group (median: 2.5, range: 0.5-9.4 nmol ml(-1)) than among the controls (median:1.1, range: 0.5 2.3 nmol ml(-1); p < 0.001) whereas paraoxonase 1 activities were lower in the ulcerative colitis group (median: 158.4, range: 61.6-264.1 U l(-1)) than in the control group (median: 233.3, range: 114.4-431.0 U l(-1); p < 0.001). There was no correlation between serum malondialdehyde level, paraoxonase 1 activity and disease activity. (1) Increased reactive oxygen metabolites levels in ulcerative colitis may result in a pro-oxidation environment, which in turn could result in decreased antioxidant paraoxonase 1 activity and increased malondialdehyde levels, (2) increased cytokines may be a possible cause of decreased paraoxonase 1 activity and (3) decreased serum paraoxonase 1 activity may be a part of an inflammatory response. PMID- 15830399 TI - Proficiency testing for cytologists. PMID- 15830400 TI - Proficiency testing: planning for the future. PMID- 15830401 TI - An open study of risperidone liquid in the acute phase of schizophrenia. AB - An open-label study was performed to investigate the clinical efficacy and mechanisms of risperidone liquid in ameliorating positive symptoms in the acute phase of schizophrenia. Eighty-eight patients (M/F: 50/38; age: 18-74 years;, mean +/- SD =32 +/- 16 years) meeting DSM-IV criteria for schizophrenia and treated with risperidone liquid (14 patients also used lorazepam) were evaluated with regard to their clinical improvement and extrapyramidal side effects using the positive and negative syndrome scale (PANSS) and the Simpson and Angus scale (SAS), while plasma concentrations of HVA and MHPG were analysed by HPLC-ECD before and 4 weeks after risperidone liquid administration. Patients showing a 50% or greater improvement in PANSS scores were defined as responders. An improvement in the PANSS scores related to excitement, hostility and poor impulse control was seen within 7 days after administration of risperidone liquid, and an improvement with regard to hallucinatory behaviour and uncooperativeness was seen within 14 days after its administration. Finally, 68% of patients were classified as responders 4 weeks after risperidone liquid administration. The scores of SAS were not changed after risperidone liquid administration. Pretreatment plasma homovanillic acid (HVA) levels in the responders (8.1 +/- 2.9 ng/ml) were higher than those in nonresponders (5.9 +/- 1.9 ng/ml). In addition, a negative correlation was seen between the changes in plasma HVA levels and the percentage of improvement in PANSS scores. On the other hand, there were no differences between pretreatment plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) levels and those of nonresponders. These results suggest that risperidone liquid is effective and well tolerated for the treatment of acute phase schizophrenic patients, and that efficacy is related to its affects on dopaminergic activity, not noradrenergic activity. PMID- 15830402 TI - The effect of quetiapine on aggression and hostility in patients with schizophrenia. AB - A post hoc analysis of data from three placebo-controlled, double-blind, randomized trials was carried out to determine the efficacy of quetiapine in aggression and hostility in patients with schizophrenia. Quetiapine treatment induced statistically significantly greater improvements in BPRS positive symptom cluster scores and three measures of hostility derived from the BPRS, compared with placebo, in patients symptomatic at baseline. A path analysis showed that the improvements in hostility were highly correlated with improvements in positive symptoms and there was no consistent relationship between sedation and hostility. Aggressive behaviour appears to be related to positive symptoms of schizophrenia. Quetiapine may be a suitable option for patients with schizophrenia and aggressive behaviour. PMID- 15830404 TI - From AgCN chains to a fivefold helix and a fishnet-shaped framework structure. PMID- 15830403 TI - Genetically selected cyclic-peptide inhibitors of AICAR transformylase homodimerization. PMID- 15830405 TI - Switchable supramolecular organization of structurally defined micelles based on an amphiphilic fullerene. PMID- 15830406 TI - Synthesis of enantiopure phthalides including 3-butylphthalide, a fragrance component of celery oil, and determination of their absolute configurations. AB - Enantiopure phthalides 2 and 5-8 were synthesized via enantioresolution of the corresponding alcohols with a chiral auxiliary of camphorsultam dichlorophthalic acid, (1S,2R,4R)-(-)-CSDP acid 3, followed by solvolysis with KOH in MeOH and the catalytic oxidation of chiral glycols with iridium complex 28. The absolute configurations of phthalides 2 and 5-8 were determined by applying the (1)H-NMR anisotropy method of MalphaNP acid (4), 2-methoxy-2-(1-naphthyl)propionic acid, to the chiral synthetic precursory alcohols. In the case of 3-phenylphthalide (R) (-)-7, the absolute configuration determined by the (1)H-NMR anisotropy method using MalphaNP acid 4 agreed with that by the X-ray crystallographic method. By applying these methods, 3-butylphthalide (S)-(-)-2, a fragrance component of essential oil of celery, has been synthesized in an enantiopure form, and its absolute configuration was unambiguously determined. PMID- 15830407 TI - Preparation of a chiral ferrocenyl derivative containing a biphenyl unit via nickel-mediated homocoupling. AB - Ferrocenyl derivative 2, which possesses a biphenyl unit, was prepared by nickel(0)-mediated homocoupling of a chiral ferrocenyloxazoline. Compound 2 exists as an equilibrium mixture of two diastereomers, in a ratio dependent on temperature and solvent, due to the partially hindered rotation around biaryl bond. Upon complexation with Cu(I) or Zn(II), complete fixation of the stereogenic axis was obtained and a single stable atropoisomer was formed, whose absolute configuration was assigned on the basis of its NOESY spectrum. The 2 Cu(I) complex was used as catalyst in the cyclopropanation of styrene. PMID- 15830408 TI - Perspectives. States unhappy with most of Feds' hoped-for Medicaid cuts. PMID- 15830409 TI - Historical vignettes in heart failure. PMID- 15830410 TI - [Control of hemorrhage in Wunderlich's syndrome]. PMID- 15830411 TI - [Irritable bowel syndrome]. PMID- 15830412 TI - The paths from research to improved health outcomes. PMID- 15830413 TI - bmjupdates+, a new free service for evidence-based clinical practice. PMID- 15830414 TI - Dexamethasone reduced the incidence of children with mild croup who returned for medical care. PMID- 15830415 TI - Review: beta agonist delivery by metered dose inhaler with a valved holding chamber (compared with a nebuliser) reduces admissions in preschool children and infants with acute asthma or wheezing. PMID- 15830416 TI - A multifaceted, home based, environmental intervention reduced asthma related morbidity in children with atopic asthma. PMID- 15830417 TI - Short term coseasonal sublingual immunotherapy reduced the development of asthma in children with hay fever. PMID- 15830418 TI - Fluoxetine plus cognitive behavioural therapy was most effective for adolescents with major depressive disorder. PMID- 15830420 TI - Review: vaccination reduces the incidence of serologically confirmed influenza in healthy adults. PMID- 15830419 TI - A telephone psychotherapy programme improved clinical outcomes in patients beginning antidepressant treatment. PMID- 15830421 TI - Review: antioxidant supplementation does not reduce gastrointestinal cancer. PMID- 15830422 TI - Fluconazole reduced the rate of recurrence of vulvovaginal candidiasis. PMID- 15830423 TI - Volunteers trained in CPR and use of automated external defibrillators increased survival after out of hospital cardiac arrest. PMID- 15830424 TI - Methylprednisolone was associated with an increase in death after head injury. PMID- 15830425 TI - Review: no evidence exists that thrombolysis is better than heparin for reducing the risk of recurrent pulmonary embolism and death. PMID- 15830426 TI - Review: a single dose of oral naproxen or naproxen sodium reduced acute postoperative pain in adults. PMID- 15830427 TI - Short term catheterisation after vaginal prolapse surgery increased recatheterisations, but reduced urinary tract infections, duration of catheterisation, and hospital stay. PMID- 15830428 TI - Review: magnesium prophylaxis after cardiac surgery reduces the risk of arrhythmia and atrial fibrillation. PMID- 15830429 TI - Review: non-invasive interventions improve symptoms and psychological functioning in patients with lung cancer. PMID- 15830430 TI - Review: capillary refill time, abnormal skin turgor, and abnormal respiratory pattern are useful signs for detecting dehydration in children. PMID- 15830431 TI - Review: a low clinical probability plus a normal D-dimer test result excludes a diagnosis of deep venous thrombosis. PMID- 15830432 TI - Mothers with post-traumatic stress disorder after traumatic childbirth struggled to survive and experienced nightmares, flashbacks, anger, anxiety, depression, and isolation. PMID- 15830433 TI - Parents' decisions on MMR vaccination for their children were based on personal experience rather than scientific evidence. PMID- 15830434 TI - Patients with acute exacerbations of COPD saw anxiety as a sign, rather than cause, of breathlessness. PMID- 15830435 TI - Palliative care involved a specific concept of care focusing on life and optimising patient quality of life. PMID- 15830436 TI - Arthritis symptoms, information sources, and a constantly shifting threshold of risk-benefit ratios influenced elderly patients' decisions about total joint replacement. PMID- 15830437 TI - 5 themes described the views of older people on advance care statements and their role in end of life care. PMID- 15830438 TI - [Surgical techniques: mouth cavity, epipharynx. Part VII]. PMID- 15830439 TI - Measuring sexual orientation in adolescent health surveys: evaluation of eight school-based surveys. AB - PURPOSE: To examine the performance of various items measuring sexual orientation within 8 school-based adolescent health surveys in the United States and Canada from 1986 through 1999. METHODS: Analyses examined nonresponse and unsure responses to sexual orientation items compared with other survey items, demographic differences in responses, tests for response set bias, and congruence of responses to multiple orientation items; analytical methods included frequencies, contingency tables with Chi-square, and ANOVA with least significant differences (LSD)post hoc tests; all analyses were conducted separately by gender. RESULTS: In all surveys, nonresponse rates for orientation questions were similar to other sexual questions, but not higher; younger students, immigrants, and students with learning disabilities were more likely to skip items or select "unsure." Sexual behavior items had the lowest nonresponse, but fewer than half of all students reported sexual behavior, limiting its usefulness for indicating orientation. Item placement in the survey, wording, and response set bias all appeared to influence nonresponse and unsure rates. CONCLUSIONS: Specific recommendations include standardizing wording across future surveys, and pilot testing items with diverse ages and ethnic groups of teens before use. All three dimensions of orientation should be assessed where possible; when limited to single items, sexual attraction may be the best choice. Specific wording suggestions are offered for future surveys. PMID- 15830440 TI - Women and cardiovascular disease. PMID- 15830441 TI - Adults' approval and adolescents' alcohol use. AB - PURPOSE: To compare adults' approval of adolescents' alcohol use among white, black, and Latino youth and to evaluate the effects of approval on most recent alcohol consumption, past 30-day use and binge drinking. METHODS: A cross sectional telephone survey of N = 6245 adolescents from 242 communities was conducted as part of the National Evaluation of the Enforcing Underage Drinking Laws Program. The survey assessed perceived availability of alcohol, underage alcohol use, and problems related to underage drinking. Ordinary least squares regression modeling was used to test the relationships between adults' approval and most recent consumption. Logistic regression modeling was used to measure the association among approval, past 30-day use and binge drinking. RESULTS: Perceived consequences, parent and adult relative provision of alcohol, and drinking with a parent were protective of underage drinking. Providing alcohol at a party, however, was associated with a two-fold increase in past 30-day use and binge drinking. There were minimal differences on adults' approval across the three racial/ethnic groups. CONCLUSIONS: Adults' approval of alcohol use is highly correlated with youth drinking behavior and has differential effects on adolescents' alcohol use depending on the social context in which the alcohol is provided. PMID- 15830443 TI - Gene therapy: the state of the art. PMID- 15830442 TI - Microbial transformation of (-)-isolongifolol and butyrylcholinesterase inhibitory activity of transformed products. AB - The microbial transformation of (-)-isolongifolol (1) by using the standard two stage fermentation technique with Fusarium lini afforded polar oxygenated metabolites: 10-oxoisolongifolol (2), 10alpha-hydroxyisolongifolol (3), and 9alpha-hydroxyisolongifolol (4). Metabolites 3 and 4 were also formed with the incubation of 1 with Aspergillus niger. All three metabolites were found to be new. Compounds 3 and 4 inhibited butyrylcholinesterase enzyme in a concentration dependent manner with IC50 values 13.6 and 299.5 microM, respectively. Compound 3 showed un-competitive mode of inhibition against butyrylcholinesterase with Ki value 15.0 microM. The structures of metabolites 2-4 were deduced on the basis of spectroscopic techniques and single-crystal X-ray diffraction techniques. PMID- 15830444 TI - Tackling tobacco use. PMID- 15830446 TI - Alternative to depleted uranium is carcinogenic in rats. PMID- 15830451 TI - Brazil approves research with embryo stem cells. PMID- 15830447 TI - Teens speak out about HIV/AIDS: focus group discussions about risk and decision making. AB - PURPOSE: To better understand the factors teens consider when making decisions regarding sex and condom use. METHODS: Twenty-one same-sex focus groups were conducted with a total of 92 male and female teens from a range of high school programs; schools were selected based on the range of programs and diversity of students enrolled. Focus group moderators facilitated groups using a set of open ended key questions about sexual decision-making and HIV/AIDS for each group; discussion on a specific topic area continued until no new information was generated. Group discussions were tape-recorded and transcribed, and themes were identified through independent content analysis of the transcriptions and assistant moderator notes. RESULTS: Although teens acquire HIV/AIDS knowledge in school and are aware of positive and negative outcomes of engaging in sexual behavior, many times the information does not seem salient or personally relevant and thus is not used in making decisions related to having sex. In general, results indicate that boys and girls seem to have different ideas/reasons for having or not having sex. Boys relate to the physical nature of sex, whereas girls relate to the emotional aspects. Teens report being bored with AIDS education, but suggest needing information that is more relevant for them. CONCLUSIONS: Attempts should be made to make HIV education more relevant for teens so that they use the information they have when making decisions about safer sexual behavior. Different approaches may be needed for boys and girls. PMID- 15830454 TI - Cancer immunotherapy: new sources for antitumour T cells. PMID- 15830455 TI - Stem cells an option after reconstructive surgery. PMID- 15830457 TI - Incidence and costs of accidents among attention-deficit/hyperactivity disorder patients. AB - PURPOSE: The purpose is to analyze the incidence and costs of accidents among Attention-Deficit/Hyperactivity Disorder (ADHD) patients. METHODS: The analysis relied on administrative medical, pharmaceutical, and disability claims for a national manufacturer's employees, spouses, dependents, and retirees (n > 100,000). Accidental injuries were identified using ICD-9 codes for injuries or poisoning treatment. ADHD sample consists of individuals with at least one claim for ADHD during 1996-98 (NADHD = 1308), which was compared with a matched control sample. In addition to descriptive statistics, multivariate analysis involving logistic regression was used to model the probability of having an accident claim in 1998. This probability was estimated for the whole population, for adults alone, for children (under age 12 years), and for adolescents (age 12-18 years). We also estimated a generalized estimation equation (GEE) model to account for the possibility of multiple accident claims for a single patient. RESULTS: ADHD patients had a greater probability of having at least one accident claim than their controls for children (28% vs. 18%), adolescents (32% vs. 23%), and adults (38% vs. 18%). Although ADHD patients' costs were greater than their controls for adults ($483 vs. $146), there was no difference for children or adolescents. However, among patients with accident claims, the average number of accident claims was similar for both groups (3.6 vs. 3.5) and costs were not statistically different. The multivariate analysis confirms this utilization pattern: the odds of having an accident for ADHD patients were 1.7 times greater than for controls. CONCLUSIONS: ADHD was a significant predictor of having an accident claim. However, for people with an accident claim, ADHD patients and controls had a similar number of accident claims and costs. PMID- 15830458 TI - Carcinogenicity of human papillomaviruses. PMID- 15830459 TI - Patenting the poor. PMID- 15830460 TI - Prevalence of, and factors associated with, adolescent physical fighting while under the influence of alcohol or drugs. AB - PURPOSE: To determine: (a) the prevalence of physical fighting while under the influence of alcohol or drugs, and (b) the associations among demographic factors, other risk behaviors, and physical fighting while under the influence of substances. METHODS: Cross-sectional analysis of The National Longitudinal Study of Adolescent Health (Add Health) 1994-1995, a school-based, nationally representative survey of 6504 7th to 12th graders. The dependent outcome variables of interest were: "The most recent time you got into a fight, had you been drinking?" and "Have you ever gotten into a fight when you had been using drugs?" Independent variables included: demographics, adolescent characteristics and risk behaviors, home environment, and peer substance use. Univariate and bivariate analyses, and logistic regressions, using SUDAAN, were performed for the two outcome behaviors for the overall sample (p 10 mug of 5-FC per ml were amebicidal to the avirulent strain; lower concentrations of the drug, which only affected growth slightly, significantly impaired the capacity of the cells to spontaneously encyst in stationary-phase cultures. On the other hand, the virulent strains were capable of growing in the presence of 5 FC (40 mug/ml) after an initial period of susceptibility. After a few transfers in growth medium lacking the drug, 5-FC-treated virulent amebas exhibited growth parameters typical of untreated cells. However, after successive subcultures in drug-free medium, 5-FC-treated cells lost their resistance and were again susceptible to the drug. This result suggested that the capacity of the cells to develop resistance resulted from a drug-induced mechanism. Spontaneous encystment, which was normally minimal in stationary-phase A-1 or A-3 cultures, was enhanced in A-3 but not A-1 cultures treated with 5-FC (>30 mug/ml). Results obtained from experiments to determine the effectiveness of 5-FC in protecting mice experimentally infected with either A-1 or A-3 amebas indicated that the clinical usefulness of 5-FC may be limited by the capacity of the amebas to develop resistance. PMID- 15830475 TI - Cefoxitin and cephalothin: antimicrobial activity, human pharmacokinetics, and toxicology. AB - Cefoxitin, a semisynthetic cephamycin, has been compared with the widely used parenteral cephalosporin, cephalothin, in terms of antibacterial activity, human pharmacokinetics, and toxicity. For both compounds, minimal inhibitory concentrations were within the therapeutic range against the 156 gram-positive cocci tested (except group D streptococci), but cephalothin was 8 to 20 times more active. Regarding the 313 gram-negative organisms tested, both antibiotics were of approximately equal activity against cephalothin-susceptible strains, but cefoxitin was outstandingly superior against Providencia spp. and indole producing Proteus spp., and markedly better against Serratia marcescens and Bacteroides fragilis. Against these organisms, cefoxitin but not cephalothin would be expected to be therapeutically valuable. Antibiotic activity levels in the serum and urine of 18 human volunteers after parenteral administration were higher and more prolonged in the case of cefoxitin, which had an average terminal serum half-life of about 45 min and a urinary recovery of about 90%. Cefoxitin was entirely nontoxic and, given intramuscularly, slightly less painful then cephalothin. These preliminary results suggest that cephamycins may prove to be a significant chemotherapeutic advance. PMID- 15830476 TI - Effect of probenecid on the apparent volume of distribution and elimination of cloxacillin. AB - According to Gibaldi et al. (1968, 1970), the higher serum concentrations of penicillins and cephaloridine reached after administration of probenecid are due not only to slower renal elimination but also to an altered distribution in the body. To determine whether probenecid has a direct effect on the distribution of cloxacillin, the elimination and distribution of cloxacillin was studied in six patients, five lacking kidney function and one with a partially impaired renal function, in the presence or absence of probenecid. No significant difference was found between the mean values of the volume of distribution of cloxacillin with and without probenecid (13.0 and 12.6 liters, respectively). Thus, the hypothesis of Gibaldi et al. could not be confirmed for cloxacillin in patients lacking kidney function. In spite of the absence of kidney function, the value of the elimination rate constant was significantly decreased in the presence of probenecid (from 0.326 to 0.263/h). This might be explained by a blockade by probenecid of the elimination of cloxacillin by the liver. PMID- 15830477 TI - In vivo effect of thiolutin on cell growth and macromolecular synthesis in Escherichia coli. AB - Thiolutin reversibly inhibits growth and ribonucleic acid synthesis in Saccharomyces cerevisiae. It is now demonstrated that, at 5 mug/ml, thiolutin rapidly inhibits all incorporation of radioactive precursors into ribonucleic acid and protein in Escherichia coli, although the incorporation of deoxythymidine into deoxyribonucleic acid continues for some time. Concentrations of thiolutin of 5 mug/ml and above are bacteriostatic and do not lead to unbalanced growth, so that cell size remains constant. The antibiotic and its inhibitory effects are easily removed by washing, whereupon macromolecular synthesis and cell division resume unimpeded. These data are consistent with reversible inhibition of ribonucleic acid synthesis being the primary mode of action of thiolutin in E. coli, and suggest that thiolutin may be a useful tool for studies where such reversible inhibition is required. PMID- 15830478 TI - Antimicrobial susceptibilities of anaerobic bacteria: recent clinical isolates. AB - Minimal inhibitory concentrations of clindamycin, minocycline, metronidazole, penicillin, and carbenicillin were determined by agar dilution against 150 recent clinical isolates of anaerobic bacteria. Ninety-nine percent of Bacteroides fragilis and all B. melaninogenicus, Clostridium perfringens, and Fusobacterium were inhibited by clindamycin at 3.1 mug/ml. Only 58% of other clostridial species were inhibited by this concentration of clindamycin. Minocycline at 3.1 mug/ml inhibited 72% of C. perfringens, 81% of other Clostridium species, and 66, 75, and 100% of B. fragilis, B. melaninogenicus, and Fusobacterium, respectively. Metronidazole at 12.5 mug/ml inhibited all bacteria tested. B. fragilis was resistant to both penicillin and carbenicillin at 6.2 mug/ml. Concentrations of 25 mug/ml for penicillin and 100 mug/ml for carbenicillin were needed to inhibit more than 90% of B. fragilis. Organisms other than B. fragilis were moderately or extremely susceptible to the penicillins. PMID- 15830479 TI - Comparison of enzymatic and microbiological gentamicin assays. AB - The correlation coefficient between the rapid enzymatic and the overnight microbiological assays for 211 urine and serum specimens was 0.96. The 95% confidence limits yielded a correlation coefficient between 0.92 and 0.98. Both methods tended to underestimate the amount of a gentamicin added to urine. When only serum samples were considered, the predicted value obtained from the linear regression analysis of either method was within 0.57 mug/ml 99% of the time. This high degree of positive correlation will permit safe rapid adjustment of individualized patient gentamicin dosages. PMID- 15830480 TI - Susceptibility of clinical isolates of bacteria to cefoxitin and cephalothin. AB - The susceptibility of 4,929 unselected clinical isolates of bacteria to cefoxitin and cephalothin was determined by the single-disk method, using a computer associated electronic zone analyzer to obtain, record, and process measurements of sizes of zones of inhibition. Both cefoxitin and cephalothin were effective against most gram-positive strains, including Staphylococcus aureus, S. epidermidis, micrococci, and all streptococci except enterococci. The three strains of Listeria monocytogenes tested were susceptible to cephalothin but resistant to cefoxitin. There was little difference between the cefoxitin and cephalothin susceptibility of Salmonellae, Citrobacter sp., Enterobacter sp., Proteus mirabilis, and Pseudomonas sp. Cefoxitin was more effective then cephalothin against Escherichia coli, Klebsiella sp., Serratia sp., indole positive Proteus sp., Providence sp., Flavobacter sp., Herellea vaginicola, and Mima polymorpha. Cefoxitin also appeared to exhibit enhanced activity, as compared with cephalothin, against Bacteroides sp. Thus cefoxitin appears to have a very broad antibacterial spectrum which is greater than that of cephalothin, especially against gram-negative strains. PMID- 15830481 TI - 5-fluorocytosine and amphotericin B in bronchial secretions. AB - The penetration into and clearance from bronchial secretions of 5-fluorocytosine and amphotericin B were studied in a dog model. After a single intravenous dose of 35 mg/kg, 5-fluorocytosine intrabronchial concentrations were greater than the minimal inhibitory concentration for 80 to 90% of Candida species. These inhibitory concentrations persisted up to 3 h. In contrast, amphotericin B in intravenous doses of 0.6 and 1.2 mg/kg penetrated the blood-bronchus barrier poorly. PMID- 15830482 TI - In vitro resistance of Neisseria gonorrhoeae to metronidazole. AB - Susceptibility of 64 clinical isolates of Neisseria gonorrhoeae to metronidazole was determined by the agar dilution technique using Mueller-Hinton and chocolate agar media. All isolates were resistant to metronidazole at 12.5 mug/ml in either medium. This lack of activity of metronidazole against N. gonorrhoeae would suggest that the ability to recover N. gonorrhoeae in patients being treated with metronidazole for concurrent Trichomonas vaginalis infections should not be hampered. PMID- 15830483 TI - Variation in performance of the serum bactericidal test. AB - Questionnaires were sent to 37 diagnostic microbiology laboratories to assess their methods for performance of the serum bactericidal test. Of the 28 responding laboratories, 26 utilized the test, with considerable variation among methods. Culture and dilution media included Trypticase soy, Mueller-Hinton, brain heart infusion, Columbia broth, dextrose phosphate, and pooled human serum. Several different schemes for dilution of the test serum were employed, including both micro and macro methods. The greatest variation occurred in the size of the bacterial inoculum, with only eight laboratories having a method for standardization of the inoculum. Other differences included the time of incubation, the method of subculture, and determination of the bactericidal end point. These studies emphasize the need for assessment of the variables in the performance of this test and for the development of a standard method for diagnostic laboratories. PMID- 15830484 TI - Synthesis of deactoxycephalosporin C by a mutant of Cephalosporium acremonium. AB - A number of Cephalosporium acremonium mutants blocked in the synthesis of cephalosporin C were investigated for accumulation of other beta-lactam compounds. The non-cephalosporin C producers were isolated after exposing the superior cephalosporin C-producing strain M8650-4 to ultraviolet light (268 nm). One of the blocked mutants, MH63, accumulated deacetylcephalosporin C (0.4 mg/ml), deacetoxycephalosporin C (1.5 mg/ml), and penicillin N (2.7 mg/ml). In contrast, the parent of MH63 produced high levels of cephalosporin C as well as deacetylcephalosporin C (2.2 mg/ml) and penicillin N (1.0 mg/ml), but only traces of deacetoxycephalosporin C (about 0.1 mg/ml). Deacetoxycephalosporin C was isolated from the mutant strain and identified by ultraviolet light, nuclear magnetic resonance, bioactivity spectrum, and co-migration with authentic standard in three chromatography systems. PMID- 15830485 TI - Effects of rate of infusion and probenecid on serum levels, renal excretion, and tolerance of intravenous doses of cefoxitin in humans: comparison with cephalothin. AB - Using a randomized crossover design, 1-g intravenous doses of cephalothin and cefoxitin, a cephalosporinase-resistant cephamycin, were infused into 12 normal adult males over periods of 120, 30, and 3 min, the last with and without prior intravenous infusions of probenecid (1 g). Mean peak serum concentrations of antibiotic activity after cephalothin infusions were 23, 56, 103, and 102 mug/ml, respectively, and after cefoxitin infusions they were 27, 74, 115, and 125 mug/ml, respectively. Probenecid treatment prolonged the terminal serum half-life of cephalothin-like activity from 0.52 to 1.0 h, and of cefoxitin from 0.68 to 1.4 h. In contrast to cephalothin, which was found to be metabolized about 25% to the less active desacetyl form, cefoxitin was metabolized less than 2% to the virtually inactive descarbamyl form, as judged from urinary recoveries. Neither antibiotic displayed detectable organ toxicity. Of 300 recent clinical isolates of gram-negative bacilli other than Pseudomonas spp., 83% were susceptible to cephalothin but 95% were susceptible to cefoxitin. Organisms resistant to cephalothin but susceptible to cefoxitin included strains of Escherichia coli, Proteus vulgaris, Klebsiella spp., Serratia marcescens, Enterobacter spp., and Bacteroides spp. PMID- 15830486 TI - Purification and characterization of pertucin produced by Pseudomonas pertucinogena. AB - Pertucin, a bacteriocin active against phase I organisms of Bordetella pertussis, was extracted from Pseudomonas pertucinogena, strain 190 or 6627, by freezing and thawing of bacteria-free agar medium and was purified by ammonium sulfate fractionation, dialysis, zinc chloride precipitation, and polyacrylamide gel electrophoresis. The specific activity of purified pertucin was approximately 80 times that of the crude extract. The purified pertucin preparation was homogeneous on ultracentrifugal analysis and polyacrylamide gel electrophoresis, and was found to be composed of protein (80%) and carbohydrate (20%), the latter assumed not to be bound covalently to the protein. Neither nucleic acids nor lipids were demonstrated. N-terminal analysis by dansylation revealed only methionine in strain 190 and only glycine in strain 6627. The isoelectric point was found to be 4.8, and the molecular weight was estimated to be 31,500. Amino acid analysis of the protein moiety demonstrated that it contained all of the common amino acids with somewhat more of the hydrophilic amino acids. With respect to thermostability, pH stability, and effects of metal ions and various buffers, pertucin behaves like a protein and is stable in 0.05 M tris(hydroxymethyl)aminomethane buffer, pH 8.0, containing 0.01 M Ca(2+). PMID- 15830487 TI - Inhibition of herpes simplex virus replication by phosphonoacetic acid. AB - Replication of herpes simplex virus in WI-38 cells was inhibited by phosphonoacetic acid, as measured by decreased virus cytopathogenic effect and incorporation of radiolabeled thymidine in virus-infected cells. The drug appeared to have no effect on adsorption, penetration, or release of the virus nor on the synthesis of ribonucleic acid or protein. It appeared to inhibit virus deoxyribonucleic acid synthesis. PMID- 15830488 TI - Impaired penicillin production in lysine regulatory mutants of Penicillium chrysogenum. AB - Lysine-regulatory mutants of Penicillium chrysogenum which excrete lysine were found to be deficient in production of penicillin. A revertant recovered the ability to produce penicillin. PMID- 15830489 TI - Dihydrofolate reductase in Pediococcus cerevisiae strains susceptible and resistant to amethopterin. AB - A mutant strain of Pediococcus cerevisiae was obtained which had 1,000-fold greater resistance to amethopterin than the P. cerevisiae/PteGlu strain. The dihydrofolate reductase level of the mutant was about 60-fold higher than that of the parent. The mutant accumulated folate, though less efficiently than its parent. PMID- 15830490 TI - In vitro uptake of isometamidium and diminazene by Trypanosoma brucei. AB - The cattle trypanocide, isometamidium, was readily taken up by inactivated Trypanosoma brucei, while its uptake by living parasites was reduced or inhibited by plasma. In both respects isometamidium differs from diminazene. PMID- 15830491 TI - Growth inhibition of enteric bacteria by Vibrio cholerae in nutrient media containing lactate, acetate, or citrate. AB - In cross-streak tests on nutrient agar containing lactate, acetate, or citrate, the growth of certain enteric bacteria was inhibited by themselves or by Vibrio cholerae, or by both. This mimicked the activity of bacteriocins, but the phenomenon was attributable to the accumulation of carbonate and increased alkalinity of the media. PMID- 15830492 TI - [Obstetric brachial plexus lesion]. PMID- 15830493 TI - [Primary treatment of Duchenne-Erb/Klumpke's paralysis]. PMID- 15830494 TI - [The effect of caffeine on hypertension]. PMID- 15830495 TI - [Recovery and rehabilitation in the field of psychiatry: old wine in new bottles or a new concept with fresh content?]. PMID- 15830496 TI - [Colesevelam hydrochloride. A product description]. PMID- 15830497 TI - [Regular self-examination or clinical examination for early detection of breast cancer]. PMID- 15830498 TI - [Depression during pregnancy and postpartum]. PMID- 15830499 TI - [Knowledge of warning signs of cancer with a view to early diagnosis. A qualitative interview study]. PMID- 15830500 TI - [Ropinirole treatment of restless legs]. PMID- 15830501 TI - [More native mitral valves can be preserved during non-ischemic mitral valve surgery]. PMID- 15830502 TI - [Preoperative fasting in management of orthopedic wound infections]. PMID- 15830503 TI - [Late first-trimester invasive prenatal diagnosis--secondary publication. An international randomized trial]. PMID- 15830504 TI - [Acute brachial neuropathy following influenza vaccination]. PMID- 15830505 TI - [Unexplained spontaneous bleeding from the uterine artery]. PMID- 15830506 TI - [Terbinafine-induced hyperamylasaemia]. PMID- 15830507 TI - [Acute renal failure in critically ill patients]. PMID- 15830508 TI - [Notification of a possible conflict of interest in Ugeskrift for Laeger]. PMID- 15830509 TI - [Mesalazine is effective in Crohn disease 1]. PMID- 15830510 TI - [Mesalazine is effective in Crohn disease 2]. PMID- 15830511 TI - [Metabolic syndrome--physiopathology of visceral adiposis syndrome and arteriosclerosis]. PMID- 15830512 TI - [Physiopathology and treatment of diabetic aortic disease]. PMID- 15830513 TI - [Significance and management of hyperlipemia as a risk factor for the progression of arteriosclerosis]. PMID- 15830514 TI - [Hypertension as a risk factor for arteriosclerosis--a lesson from recent, large scale clinical tests]. PMID- 15830515 TI - [Non-surgical evaluation of arteriosclerosis--carotid artery echography and pulse velocity]. PMID- 15830516 TI - [Progress in diagnosis of coronary disease: Blood markers, echocardiography and scintigraphy]. PMID- 15830517 TI - [Up-to-date intervention in acute coronary syndrome]. PMID- 15830518 TI - [Future prospect in diagnosis and treatment of coronary artery diseases]. PMID- 15830519 TI - [Diagnosis and treatment of gastroesophageal reflux disease]. PMID- 15830520 TI - [Current topics on therapy of hepatocellular carcinoma]. PMID- 15830521 TI - [Physiopathology and treatment of chronic pancreatitis]. PMID- 15830522 TI - [Diagnosis and treatment of hypopituitarism]. PMID- 15830523 TI - [Diabetes mellitus and arteriosclerosis--recent findings in non-invasive testing]. PMID- 15830524 TI - [brain diseases associated with diabetes mellitus]. PMID- 15830525 TI - [Clinical description of Parkinson's disease]. PMID- 15830526 TI - [Angiitis--the current concept and treatment]. PMID- 15830527 TI - [Facing the evolutionary period for the treatment of rheumatoid arthritis]. PMID- 15830528 TI - [Physiopathology of chronic heart failure and progress in its treatment]. PMID- 15830529 TI - [Diagnosis and treatment of acute coronary syndrome]. PMID- 15830530 TI - [Physiopathology and treatment of bronchial asthma]. PMID- 15830531 TI - [Physiopathology and treatment of chronic obstructive pulmonary diseases (COPD]. PMID- 15830532 TI - [Physiopathology and treatment of leukemia]. PMID- 15830533 TI - [Physiopathology and treatment of aplastic anemia]. PMID- 15830534 TI - [Physiopathology and treatment of glomerulonephritis]. PMID- 15830535 TI - [Evaluation of renal dysfunction and blood pressure control]. PMID- 15830536 TI - [The current status and future prospect in the prevention of hospital infections]. PMID- 15830537 TI - [Surgery for lumbar narrow canal]. PMID- 15830538 TI - [Surgical treatments for the patients with SMA tumor]. PMID- 15830539 TI - [Clinical features and outcomes in patients with asymptomatic moyamoya disease- from the results of nation-wide questionnaire survey]. AB - After induction of noninvasive diagnostic tools, asymptomatic moyamoya disease is occasionally being diagnosed. However, there is no epidemiological or clinical data concerning this condition because of its rarity. To elucidate clinical features and outcomes in patients with asymptomatic moyamoya disease we conducted a nation-wide questionnaire survey. Questionnaires were sent to 88 neurosurgical institutes in 1994 and we received answers from 66 institutions (75% recovery). Diagnosis of moyamoya disease is based on the guidelines established by the research committee on moyamoya disease of the Ministry of Health and Welfare, Japan. Thirty three asymptomatic moyamoya disease patients were collected (11 male, 22 female) and divided into 2 groups; group 1 defined as patients without any symptoms, and group 2 as patients who did not show any symptoms except headache. 80% of group 1 patients were adults, whereas in group 2, children and adults were equally distributed. Follow-up periods were 3 years and 8 months on average. Among conservatively treated 28 patients, 2 patients died from suspected bleeding (mortality 7%) and 4 patients with TIAs resulted in good outcome. Among surgically treated 5 patients, 1 patient showed poor outcome due to post operative infarction. Other 26 patients showed excellent outcome. Natural course of asymptomatic moyamoya disease seemed benign, but mortality from bleeding is not negligible. A prospective study of asymptomatic moyamoya disease is necessary to clarify its natural course and appropriate management strategy. PMID- 15830540 TI - [Intraoperative radical clot removal therapy using a bipolar irrigation system for prevention of cerebral vasospasm following subarachnoid hemorrhage]. AB - To investigate cerebral vasospasm following subarachnoid hemorrhage (SAH), 65 cases were analyzed by the incidence of delayed ischemic neurological deficit (DIND). Eighty-four consecutive cases, who underwent surgery within 72 hours of SAH, were retrospectively analyzed. Sixty-five of these patients had diffuse subarachnoid hemorrhage in CT scan and had aneurysms of the anterior circulation. They were divided into two groups according to the operative procedure: Group A (35 cases) had radical clot removal therapy with urokinase in 5% glucose solutions using a bipolar irrigation system; Group B (30 cases) had conventional clot removal using manual irrigation. There were no significant differences between these two groups for age or pre-operative WFNS grade. The incidence of DIND was 17.1% in group A and 40.0% in groupB; this was a significant difference. Radical clot removal therapy was effective in preventing DIND due to vasospasm. PMID- 15830541 TI - [Three-dimensional imaging of the optic nerve using magnetic resonance angiography--application to anterior communicating artery aneurysm and craniopharingioma]. AB - The purpose of this investigation was to analyze three-dimensional images of the optic nerve obtained by magnetic resonance angiography (MRA) in cases of anterior communicating artery aneurysm and craniopharingioma. Four ruptured anterior communicating artery aneurysms, five non-ruptured anterior communicating artery aneurysms and two craniopharingiomas were examined. The images were taken using MR/i Hispeed Plus 1.5T Infinity version, and analyzed by Advantage Work station AW4.1. The routine MR imaging parameters are shown in Table. The imaging time was about 10 minutes. Analysis was made by reformation of images parallel to the optic nerve obtained from the original MRA images. The optic nerve and brain tumor were traced with paintbrush from one sheet to another of the reformed images after subtraction of the blood vessels around the anterior communicating artery in these reformed images, and then three-dimensional images were constructed. Three-dimensional images of the blood vessels were reconstructed from MIP (maximum intensity projection) images using the threshold method. The optic nerve and anterior communicating arterial aneurysm or brain tumor were both observed in the overlapped 3D-SSD (shaded surface display) images. The analysis time was about 15 minutes. Three-dimensional images of the optic nerve and anterior communicating artery aneurysm or brain tumor were able to be made in all cases. As a preoperative investigation for anterior communicating artery aneurysm or suprasellar brain tumor, we considered that three-dimensional imaging of the optic nerve is useful in the operative approach because the optic nerve acts as a merkmal for the anterior communicating aneurysm or brain tumor. PMID- 15830542 TI - [Craniotomy for organized subdural hematoma]. AB - We reported 3 cases of organized chronic subdural hematoma (CSDH), which required radical treatment by craniotomy. All patients were in the 70s (male, two; female, one). Two cases had previously received craniotomy and neck clipping for intracranial aneurysm. All of them received trephine evacuation of hematoma more than two times, although the hematoma was not removed due to organization. Because of this, we performed craniotomy and removal of the hematoma with its thick outer membrane. In order to obliterate the subdural space completely, we stripped the dura mater from the skull and attached it to the inner membrane with fibrine glue. After a period of more than two years of followed-up, no recurrences had been discovered. PMID- 15830543 TI - [A case report of ruptured internal carotid artery aneurysm with contralateral oculomotor nerve paresis]. AB - We report a case of ruptured internal carotid artery aneurysm with contralateral oculomotor nerve paresis. A 69-year-old female experienced a sudden onset of severe headache. Left-sided mydriasis, absence of light reflex, disability of left ocular movement and ptosis were identified. Computed tomography scan revealed massive subarachnoid hemorrhage. Cerebral angiography showed a right internal carotid artery aneurysm. No abnormal finding was observed in the left internal carotid artery or basilar artery. Magnetic resonance imaging did not show thrombosed aneurysm. The oculomotor nerve paresis lasted for ten days. Ruptured right internal carotid artery aneurysm was surgically confirmed and repaired. We speculated that the contralateral oculomotor nerve paresis was the consequence of bloody jet flow from the ruptured aneurysm. PMID- 15830544 TI - [Extracranial vertebral artery dissection repeating distal embolism: case report]. AB - We report a case of vertebral artery (VA) dissection presenting with repeated distal embolism. A 48-year-old man was admitted to our institution with neck pain and vertigo of two day duration. MRI on arrival showed infarction of the left cerebellar hemisphere and the right occipital lobe. Emergency angiography revealed left VA dissection and occlusion of the left posterior inferior cerebellar artery and right calcaline artery. Six hours after admission, he manifested cortical blindness due to embolism of the contralateral left calcaline artery despite giving heparin and aspirin. Thrombolysis was performed but was not successful. We embolized the affected VA to prevent further embolism. We discuss here the indication and timing of treatment for VA dissection presenting with ischemic symptom. PMID- 15830545 TI - [Coil embolization with a hyperform occlusion balloon for an acutely ruptured wide-necked basilar tip aneurysm: case report]. AB - A 54-year-old woman was admitted to our hospital because of severe headache. CT scan demonstrated subarachnoid hemorrhage (SAH), and cerebral angiography revealed a wide-necked basilar tip aneurysm with a bleb and a small basilar artery-left superior cerebellar artery aneurysm. She was treated with coil embolization using a neck remodeling technique one day after SAH. Guiding catheters were introduced into the bilateral vertebral artery via the transfemoral route. A Hyperform occlusion balloon catheter was introduced into the left posterior cerebral artery (PCA) through the left vertebral artery, and was placed at the neck of the basilar tip aneurysm for neck remodeling. Then, the microcatheter for coil deployment was placed in the dome of the aneurysm through the right vertebral artery. The coil was deployed in the aneurysm while the balloon was inflated across the neck. The inflated Hyperform occlusion balloon was partially herniated into the dome of the aneurysm. The distal part of the balloon was positioned in the left PCA segment, and its proximal part was positioned in the basilar artery. The balloon conformed easily to the aneurysmal neck and its surrounding vessel structure. When the balloon was deflated, the coil did not protrude into the parent artery, and was stable in the aneurysm. Angiography immediately after the procedure showed complete occlusion of the aneurysm with patency of both PCAs. The Hyperform occlusion balloon is very compliant and supple, so it easily changes from its cylindrical shape when inflated to expand into the origin of the arterial branches emerging from the aneurysm neck. We think this balloon is a useful device for treating wide-necked aneurysms located at arterial bifurcation. PMID- 15830546 TI - [Ruptured aneurysm of bihemispheric anterior cerebral artery bifurcation: case report]. AB - It is well known that distal anterior cerebral artery (ACA) aneurysms are often associated with the anomalies of the ACA, of which azygos ACA and triple ACA have frequently been described. In this report, we present a rare case with a ruptured aneurysm arising from the bifurcation of the bihemispheric ACA, a rare anomaly of the ACA. A 63-year-old male suddenly developed severe headache, consciousness disturbance, and left hemiparesis, and was admitted to our hospital. Plain CT scans revealed subarachnoid clots that were densely distributed in the supracallosal cistern. Cerebral angiography demonstrated that the left pericallosal artery supplied blood flow to the bilateral parietal lobes through the bihemispheric arteries. A saccular aneurysm was found at their bifurcation. He underwent clipping surgery through interhemispheric approach. Postoperative course was uneventful. Special care should be taken not to occlude the bihemispheric arteries supplying to the bilateral parietal lobes during surgery. PMID- 15830547 TI - [A case of organized chronic subdural hematoma showing an early recurrence after craniotomy]. AB - A 64-year-old man who had undergone single burr hole drainage twice prior to this admission was hospitalized with a recurrent right chronic subdural hematoma. A head CT showed a mixed density subdural hematoma on the right frontotemporoparietal region. Based on the intraoperative findings of the previous surgeries, the hematoma was known to be organized. Therefore, we decided to do a small craniotomy under general anesthesia, and remove the organized subdural hematoma and thick outer membrane while leaving the thickened dura matter intact. The inner membrane was left untouched. One week later, despite adequate decompression, the hematoma recurred with midline shift on head CT. It is likely that the uniquely thick and vascular enriched outer membrane and dura contributed to such an early recurrence. Finally, we performed an extensive craniotomy, removing all the organized hematoma, outer membrane and dura. Again, the inner membrane was left intact. On one year follow-up the patient has been asymptomatic with complete resolution of the subdural hematoma on CT scan. The successful treatment of organized chronic subdural hematoma can be challenging. We strongly recommend an extensive removal of the organized hematoma, outer membrane and excision of the dura mater in order to achieve a successful outcome after failed burr hole evacuation. PMID- 15830548 TI - [The retina--its structure and function that are different from a digital camera]. PMID- 15830549 TI - [Ureterolysis: technique, indications]. AB - Ureteral obstruction due to idiopathic retroperitoneal fibrosis is a rare but severe clinical problem. The open approaches, as well as surgical techniques used to prevent stenosis recurrence, are described. Ureterolysis remains the procedure to relieve ureteral obstruction. The ureter is dissected and freed from the fibrotic process, and then separated to prevent the recurrence of the stenosis. Recently, the development of Laparoscopic urology has allowed for minimal invasive treatment of many urological problems. We present our technique of ureterolysis for extrinsic ureteral obstruction. Advantages and complications of each method are considered and indications are proposed. PMID- 15830550 TI - [Surgery for female stress urinary incontinence with transobturator tape]. AB - The aim of suburethral transobturator suspension is to cure the women stress urinary incontinence. The concept underlying this apparatus is based on several points: it reproduces the urethral fascia; it complies with Delancey's concept; it consists of a tension-free band through the soft structures of the obturator fossa; it is a perineal surgery. This surgery needs specific devices: a synthetic tape and a specific tool to introduce it, the tunnelling device. Respecting some technical landmarks are mandatory to ensure successful intervention: the vaginal incision must include all the thickness of the vaginal wall; the trans-obturated endpoint must be located at the level of the mid urethra; the tunnelling device must have a close contact with the ischiopubic bone; the finger inside the incision protects the urethra and drives the tunnelling device inside the vaginal incision. PMID- 15830551 TI - [Infective lithiasis]. AB - "Infection Lithiasis" refers to calculi that occur with persistent urinary tract infection. Stones composed of magnesium ammonium phosphate (struvite) and carbonate apatite, called "triple phosphate" stones, are the more common type of infection lithiasis. These stones are also called "staghorn" calculi because they may grow rapidly and fill the entire collecting system. They form during urinary infection with urea-splitting micro-organism. They may originate de novo or complicate a lithiasis when pre-existing stones are colonized with urea-splitting bacteria. They represent about 2-3% of stones referred for laboratory analysis. This article reviews the epidemiology, pathogenesis, clinical features, and management of struvite stones. A singular pathologic entity recently described, called "encrusted cystitis or encrusted pyelitis", mainly caused by Corynebacterium urealyticum is also review. Infection lithiases caused by non urease-producing bacteria may also occur and are examined in this article. Finally, the controversial role of nanobacteria in nephrolithiasis is discussed. PMID- 15830552 TI - [Upper urinary tract: physiology, pathophysiology of obstructions and function assessment]. AB - The urine is transported from the renal papilla to the bladder through the upper urinary tract which allows this transport to be safe and comfortable, i.e., without any risk or pain for the kidney. This active transport depends on the smooth muscle contractile properties. The upper urinary tract is totally autonomous; this feature allows the preservation of its function after renal transplantation. However, despite its accessory role, the autonomous nervous system can modulate its activity. Upper urinary tract obstruction involves adaptative mechanisms which are different depending on the type (acute, chronic, acquired or congenital) of obstruction. Functional evaluations of the upper urinary tract are aimed at identify the urine transport conditions and the relationship between obstruction and clinical conditions such as hydronephrosis, pain or impaired renal function. PMID- 15830553 TI - Disability. PMID- 15830554 TI - Introduction to a special section on disability ethics. The Journal of Clinical Ethics. PMID- 15830555 TI - Brain trauma and surrogate decision making: dogmas, challenges, and response. PMID- 15830556 TI - Depolarizing and complicating the ethics of treatment decision making in brain injury: a disability rights response to Nelson and Frader. PMID- 15830557 TI - Ethical issues in rehabilitation in the home-care setting. PMID- 15830558 TI - Clinical (mis)judgments of quality of life after disability. PMID- 15830559 TI - Acquired brain injury: reflections of two professionals with ABI. PMID- 15830560 TI - The trauma of discharge planning following brain injury. PMID- 15830561 TI - Hypothetical autonomy and actual autonomy: some problem cases involving advance directives. PMID- 15830562 TI - Surrogates' decisions regarding CPR, and the fallacy of substituted judgment. PMID- 15830563 TI - Feeding patients with advanced dementia: a Jewish ethical perspective. PMID- 15830564 TI - Cadaver dissection and the limits of simulation. PMID- 15830565 TI - Comment on Ganzini and Dobscha regarding comparing rates of physician-assisted suicide in Oregon with that of other states. PMID- 15830566 TI - Low-iodine diet before radioiodine uptake scans or therapy--flawed advice to U.K. patients. PMID- 15830567 TI - Anaplastic thyroid cancer in British Columbia 1985-1999: a population-based study. AB - AIMS: To review the outcome of patients diagnosed with anaplastic thyroid carcinoma in British Columbia between January 1985 and December 1999. MATERIALS AND METHODS: Seventy-five patients were identified. Survival curves were calculated using Kaplan-Meier estimates, and the charts of the 62 patients referred to a British Columbia Cancer Agency (BCCA) facility were reviewed. RESULTS: All cases: 51 out of 75 patients (68%) were women; median age of all patients was 74 years. The overall- and cancer-specific 5-year survival rates for the whole group were 5%. non-referred cases: nine out of 13 patients were women; median age at diagnosis 84 years. Eleven of the 13 patients died within 1 month of diagnosis. Referred cases: 42 out of 62 patients were women; median age 72 years; median survival 5.1 months; 1-year survival 19%. Forty-eight patients presented with tumours that extended through the thyroid capsule, 10 presented with distant metastases. Four patients had a total thyroidectomy, nine a partial thyroidectomy, and 49 a biopsy only. Fifty-seven patients received radiotherapy, nine of these had concurrent chemotherapy: Thirty-three patients received less than 40 Gy and 24 patients received 40 Gy or more. Median survival was longer (9 vs 3 months) in patients receiving 40 Gy or more; this group included four patients who had prolonged survival. CONCLUSION: Long-term, disease-free survival was achieved in a few patients who were able to receive high-dose radiotherapy, preferably after adequate surgery. PMID- 15830568 TI - Can cisplatin and infused 5-fluorouracil be replaced by oxaliplatin and capecitabine in the treatment of advanced oesophagogastric cancer? The REAL 2 trial. PMID- 15830569 TI - Review of second-line chemotherapy for advanced gastric adenocarcinoma. AB - Palliative chemotherapy has been shown to improve survival compared with best supportive care alone in patients with unresectable or recurrent gastric cancer. However, patients receiving chemotherapy eventually develop progressive disease. At this stage, no standard second-line chemotherapy can be offered. No randomised controlled trial data suggest a benefit of second-line chemotherapy compared with supportive care alone. We review the published data concerning the use of second line chemotherapy in gastric adenocarcinoma. Response rates to second-line therapy in phase II trials are similar to those seen for other cancers that are more commonly retreated. In addition, data suggest that patients who respond to second-line therapy consistently survive longer compared with non-responders, and, perhaps more importantly, symptomatic benefit may be obtained from second line therapy. PMID- 15830570 TI - Post-operative radiochemotherapy for gastric cancer: adoption and adaptation. AB - AIMS: Intergroup study 0116 (INT-0116) showed an 11% absolute improvement in 3 year survival with post-operative radiochemotherapy for gastric cancer, but reported 33% severe acute GI toxicity using conventional simulation with large fields. We adapted the treatment using conformal radiotherapy techniques and assessed toxicity and outcome in 20 consecutive patients. METHODS: A conformal radiotherapy technique previously developed for gastric lymphoma was adapted to treat the target volume defined in INT-0116. The five-field plan used a large anterior field, plus asymmetrically matched upper AP:PA fields and lower lateral fields. Consecutive patients with ECOG PS 0-2 and stage IB-IV non-metastatic gastric cancer were treated with 5-FU (425 mg/m2 daily x 5 days) and leucovorin (20 mg/m2 daily x 5 days) for one cycle prior to and two cycles following concurrent radiation (45 Gy/25 fractions) with identical drug dosages on the first 4 and last 3 days of radiation. Acute toxicity was prospectively recorded weekly using RTOG and NCI common toxicity criteria. Patient charts were reviewed in November 2003 and late toxicity and outcome were recorded. RESULTS: Nineteen of 20 patients completed radiotherapy and 14 completed all chemotherapy cycles. One patient died of neutropenic sepsis. Maximum acute toxicity [grade (number)] was: 5(1), 4(0), 3(4), 2(10), 1(4), 0(1). There were two grade 1 late toxicities. Two-year overall survival is 70% (95% confidence interval: 50-90). CONCLUSIONS: Conformal radiotherapy may improve acute toxicity (25% grade 3 or greater toxicity as compared with 41% reported in INT-0116). Survival is comparable to that achieved in the INT-0116 treatment arm (approximately 60% at 2 years). INT 0116 results can be achieved outside a study setting; however, further efforts to improve treatment efficacy and minimize toxicity are warranted. PMID- 15830571 TI - Clinical benefit of intermittent administration of imatinib in a patient with metastatic gastrointestinal stromal tumour. AB - Gastrointestinal stromal tumours (GISTs) are mesenchymal tumours of the gastrointestinal tract, resistant to conventional chemotherapy and radiotherapy, but responsive to imatinib. Acquired resistance to imatinib has been reported in some patients, and several studies have suggested the possibility of overcoming imatinib resistance by increasing the dose of the drug. This case shows the possibility of obtaining clinical benefit with intermittent administration of imatinib in a patient who developed acquired resistance to a high dose of the drug. PMID- 15830572 TI - Reduction of radiotherapy-induced late complications in early breast cancer: the role of intensity-modulated radiation therapy and partial breast irradiation. Part II--Radiotherapy strategies to reduce radiation-induced late effects. AB - Radiotherapy after conservation surgery has been proven to decrease local relapse and death from breast cancer, and is now firmly established in the management of early breast carcinoma. Currently, the challenge is to optimise the therapeutic ratio by minimising treatment-related morbidity, while maintaining or improving local control and survival. The second part of this review examines the role of two approaches: intensity-modulated radiation therapy (IMRT) and partial breast irradiation, as means of improving the therapeutic ratio. Discussion of IMRT includes both inverse- and forward-planned methods: the breast usually requires minimal modulation to improve dose homogeneity, and therefore lends itself to simpler forward-planned IMRT techniques; whereas inverse-planned IMRT may be useful in selected cases. There are many dosimetry studies reporting the superiority of IMRT over conventional breast radiotherapy, but there is still a paucity of clinical data regarding patient benefit from these techniques. A critical literature review of clinical partial breast radiotherapy studies focuses on the influence of irradiated breast volume, dose and fractionation, and patient selection on normal tissue side-effects and local control. Clinical reports of partial breast irradiation show several encouraging, but some concerning results about local recurrence rates. Therefore, mature results from randomised trials comparing partial breast irradiation with whole-breast radiotherapy are required. Accurate localisation of the tumour bed and application of appropriate clinical target volumes and planning target volumes are discussed in detail, as these concepts are fundamental for partial breast irradiation. PMID- 15830573 TI - The effect of locoregional recurrence on survival and distant metastasis after conservative treatment for invasive breast carcinoma. AB - AIMS: Patients with invasive breast cancer submitted to conservative treatment must be followed for a long period of time to study locoregional control. In this study, we analysed the outcome and relationships between locoregional recurrence (LRR), distant metastases and survival. MATERIALS AND METHODS: A 15-year study, including 470 women with early breast cancer, stage I and II, who underwent breast conservative treatment. Tumour size, nodal status, age, menopausal status, histological grade and LRR were analysed for their ability to predict overall survival, disease-specific survival and distant disease-free survival. RESULTS: With a median follow-up time of 6.6 years (3 months to 19.1 years), there were 19 LRR at their first site of recurrence and 53 distant metastases. Tumour size greater than 2 cm, positive lymph nodes and histological grade III were significantly related to lower overall and distant metastases-free survival. On multivariate analysis, nodal status, histological grade III and LRR (coded as a time-dependent variable) were significantly related to overall, specific and distant metastases-free survival, whereas tumour size had only a borderline effect on specific and distant disease-free survival. Landmark analysis showed that women who presented an LRR within 2 years after surgery had significantly lower distant disease-free survival (hazard ratio [HR]: 8.39; 95% CI 2.56-27.47; P < 0.001), specific survival (HR: 8.19; 95% CI 2.45-27.41; P < 0.001) and overall survival (HR: 6.02; 95% CI 2.25-16.11; P < 0.005). CONCLUSIONS: LRR seems to be a significant predictor of distant metastases and survival, and patients who sustain early LRR tend to display a more aggressive clinical course. PMID- 15830574 TI - The successful treatment of chylous effusions in malignant disease with octreotide. AB - Chylous effusions in adults are commonly associated with malignant disease. Although the condition is rare, their occurrence presents a significant management problem. A review of the literature demonstrates the high mortality of this condition in the past from cachexia and infection or after surgical attempts at correction. The first report of somatostatin use in chylous effusions was a decade ago. Since 2000, case reports of successful treatment in infants and neonates with intravenous somatostatin or octreotide have been published. For adults, few reports exist. We describe a case series of seven patients, all with malignancy. In each case, there was a systematic approach to treatment using subcutaneous octreotide and a fat-free diet, resulting in complete resolution of the condition. Although no guidelines are available for the management of chylous effusions, our non-invasive approach avoided lymphangiogram, surgery and allowed early discharge. PMID- 15830575 TI - Topical phenylephrine in the treatment of radiation-induced faecal incontinence. AB - AIMS: Acute bowel toxicity after pelvic radiotherapy is defined as occurring within 3 months of the start of treatment; chronic gastrointestinal toxicity may continue after the acute phase or start after a latent period. One in five patients develop chronic faecal incontinence affecting quality of life; how best to treat these patients is not known. This retrospective study aimed to determine the effects of a new agent, phenylephrine gel, in the treatment of chronic radiation-induced faecal incontinence. MATERIALS AND METHODS: Patients prescribed phenylephrine gel for new-onset faecal incontinence after radiotherapy were identified from our database of patients treated in a specialist radiation induced bowel damage clinic since 2000. Changes in the level of faecal incontinence were assessed using the Vaizey faecal incontinence scoring system before and after treatment. RESULTS: Fifteen patients (nine men and six women) of mean age 70.5 years (standard deviation 8.2, age range 56-82 years) were treated with phenylephrine gel a median of 43 months after completing radiotherapy. The median Vaizey score before treatment with phenylephrine gel was 17 (interquartile range [IQR] 14-20) and after treatment was 14 (IQR 11-18) (P = 0.005). The median length of treatment with phenylephrine gel was 28 days (IQR 28-365). Scores improved in 11 out of 15 patients; four out of 15 patients showed substantial improvements of 7 or more points; and seven patients considered the gel helpful. CONCLUSION: Topical phenylephrine gel for the treatment of radiation-induced faecal incontinence has not been previously reported. This small, retrospective study suggests that it may help most patients and, in some, the improvement may be substantial. However, larger placebo-controlled prospective studies are required. PMID- 15830576 TI - Long-term outcomes and morbidity after I125 brachytherapy for localised prostate cancer: an early U.K. series. PMID- 15830577 TI - Use of a telephone advice line for oncology patients. PMID- 15830578 TI - Patient selection for palliative whole-brain radiotherapy based on RTOG recursive partitioning analysis. PMID- 15830579 TI - Spindle-cell liposarcoma: a rare variant of liposarcoma arising in the foot. PMID- 15830580 TI - What does a good quality of life for cancer patients mean on a numerical rating scale? PMID- 15830581 TI - Early use of dexamethasone in malignant spinal-cord compression: a missed opportunity? PMID- 15830582 TI - Metastatic myxoid liposarcoma: aggressive multimodality management. PMID- 15830583 TI - Genetic analysis of modern and historical burned human remains. AB - Burning of corpses is a well-known funeral procedure that has been performed for a long time in many cultures. Nowadays more and more corpses are burned in crematories and buried in urns, often for practical and financial reasons. In some scientific, criminal or civil cases even after cremation there is the need of genetic investigations for identification or paternity testing. Furthermore, burned remains are the only remains left in North Europe from 1200 BC to 500 AD. This makes genetic investigation of those materials interesting for anthropological reasons. We present on one hand a systematic investigation of 10 corpses before and after the cremation and on the other hand the analysis of seven historical remains representing the bronze age. We chose the ground bone powder and the less destroyed bone parts respectively and employed a slightly modified commercially available DNA extraction method. The presence of human nuclear and mitochondrial DNA was tested by a simple but highly sensitive Duplex PCR. DNA quantification was done using real time PCR, and genetic typing was tried out using the AmpFISTR Identifiler Multiplex Kit, followed by an automatic analysis on an AbiPrism310. PMID- 15830584 TI - Frequency analysis of the delta32ccr5 HIV resistance allele in a medieval plague mass grave. AB - The 32 basepair deletion in the gene for the human chemokine receptor CCR5 (delta32ccr5) conferring resistance against HIV-1 infection is present in Caucasian populations. The mutant allele is believed to have originated by a single mutational event in historic times and to have reached its present population frequency of an average 10 % in Europe through selective pressure by a pathogenic agent. Because of their great impact on European populations, the medieval Plague epidemics have been considered as a possible candidate. To test this hypothesis, we studied the delta32ccr5-frequency in 35 individuals from a mass grave containing victims of the 14th century Plague pandemic in Lubeck, Northern Germany, and compared them to the frequency in a control group from the same burial site, dating from the time before the first Plague pandemic. If the delta32ccr5 allele conferred an at least partial resistance against the medieval Plague, its frequency would be expected to be lower in those that died in the pandemic, than it was in the local population before the arrival of the Plague. The CCR5 locus could be typed successfully for 14 Plague victims and for 20 individuals from the medieval control group. We found a delta32ccr5 allelic frequency of 14.2% and 12.5%, respectively. The difference between these figures is not statistically significant. Furthermore, they are comparable to the delta32ccr5 frequency for nowadays Northern Europe. We therefore conclude that the medieval Plague pandemic has not contributed to an increase in the allelic frequency of the mutant delta32ccr5 allele and that, if there has been a positive selection of this allele, it is likely to have occurred before the 14th century and thus before the arrival of the Plague in Europe. PMID- 15830586 TI - Sex and gender differences in secular trend of body size and frame indices of Lithuanians. AB - In Lithuania, as in many other European countries, positive secular trend of body size became especially expressed at the middle of XX. century. The analysis showed the evident secular trend in height of adult Lithuanians since the end of XIX. century till 2001 from 163.5 till 181.3 cm in males and from 153.3 cm till 167.5 cm in females. Concerning growth and development of children, during the 1925-2001 period especially obvious acceleration of height was determined at the middle of adolescence due to much earlier sexual maturation at the end of XX. century. Secular trend of body height was more expressed in males of all ages, nevertheless this process stabilized in both sexes during the last decades. While comparing secular changes in BMI of adolescent Lithuanian girls and boys between the 1925-2001 period, the evident sexual differences in secular changes were determined from the middle of pubescence: the course of changes in BMI during the XX. century was nearly the same in both sexes, but at the end of XX. century older adolescent girls became obviously thinner compared with the changes of BMI in boys of the same age. A very similar trend in BMI of adult Lithuanian women since 1980 till 2002 was also detected. This phenomenon could be more related with the changes in life stile, modern-day canons for unrealistically thin female's body and strong social pressure on women today. PMID- 15830585 TI - Study on the distribution of the MSY2 polymorphism in 9 Chinese populations. AB - The distribution of the MSY2 polymorphism in Chinese populations was analyzed by PCR. The results showed that the MSY2*4 allele, whose frequency in the total material was found to be 94.95%, was the common allele, while the distribution of the MSY2*3 allele was significantly different in the 416 tested males from the 9 populations under study. Based on the chi2-analysis, a distinct diversity was found in the non-group populations. Diversities were also discovered between southern and northern groups and among southern groups. On the contrary, no difference concerning the diversity was detected among the northern populations. In conclusion, MSY2 proved to be an important genetic marker with regard to the study of the genetic structure of Chinese populations, and further evidence is given concerning the migration direction between South and North. PMID- 15830587 TI - Secular changes are different in distinct subgroups of the growing population. AB - The change of living conditions in East Germany after the German reunification in 1990 led to intensive secular changes in growth and development. Anthropometric data of height, weight, BMI and thickness of the subcutaneous fat layer are compared from two cross-sectional samples of German children and adolescents aged 6 to 18 years. The first sample was measured around 1989 and the second 10 years later around 1999. Both samples contain children and adolescents from the urban as well as from the rural population. The secular changes reported here are based on the comparison of medians of height, weight, BMI and subcutaneous fat layer thickness calculated for yearly age categories. Subsequently, medians of the 10% border categories are compared. These border categories contain the 10% smallest, lightest, slimmest or leanest subjects and the 10% tallest, heaviest, most corpulent or most obese subjects respectively of a yearly age category. The young generation shifted between 1989 and 1999 to a taller stature in the small as well as in the tall category. The secular pattern of measurements, which mark corpulence, is sex-specific and differs from that of length measurements. In general the light or lean subjects changed only little, however, there are clear age- and sex-specific changes in the upper border categories. Weight and BMI increased markedly in the upper border categories in young preadolescent children, but did not change much in adolescent boys of these category aged 15 and 16 and decreased in adolescent girls. Also the subcutaneous fat layer increased in the upper border category in preadolescent children, but decreased in adolescent boys and girls. PMID- 15830588 TI - Assessment of developmental age: cross-sectional analysis of secondary sexual characteristics. AB - The developmental age of a growing person does not necessarily correspond to his or her chronological age. The two sexes differ considerably in their developmental tempo, and there are tempo differences also within the same sex. Early developers appear older, late developers appear younger than their chronological age might suggest. Based on a sample of 8675 German boys and 8689 girls of the same ethnicity, aged between 8 and 17 years sex differences of height, weight and secondary sexual characteristics are analyzed. Girls on average, develop faster than boys. Their peak of highest increments of height occurs at 10.2 years and is positioned early within the events of sexual maturity. The peak of highest increments of height in boys occurs at age 11.2 on average and is positioned relatively early within the sequences of secondary sexual characteristics. Maturing development starts with breast-stage 2 in girls at age 10.9 and penis-stage as well as scrotum-stage 2 in boys at age 11.1 on average. The development of pubic hair follows and menarche in girls at age 12.7, respectively spermarche in boys at age 13.8, marks the border to theoretical fertility. There are remarkable differences in the tempo of sexual development between the different types of body shape. In girls the pyknomorphic types are the early developers. Breast stage 2 for instance occurs at age 10.1 in the stocky and corpulent pyknomorphic girls and at age 13.0 in the tall and slender leptomorphic girls. This is different in boys. Here are much less differences between the types of body shape, but generally it is the leptomorphic type, at whom the stages of his secondary sexual characteristics are developed a little earlier. PMID- 15830589 TI - Body build classification for ordinary schoolgirls (aged 7-18 years) and volleyball girls (aged 13-16 years). AB - The article describes two Estonian anthropometric cross-sectional studies of 1549 ordinary schoolgirls (aged 7-18) and 46 girls, who regularly practised volleyball (aged 13-16). Data are presented on 22 basic anthropometric measurements and 6 body composition characteristics (body mass index, mean skinfold, body density, relative mass of fat by Siri, absolute mass and relative mass of subcutaneous adipose tissue). All anthropometric variables were classified into five height weight SD classes. Schoolgirls were divided into six age groups (7-8, 9-10, 11 12, 13-14, 15-16, 17-18). Volleyballers were observed as one group as their age in SD classes did not differ significantly. The classification consisted of five categories: three height-weight concordant categories: I--small (small height, small weight), II--medium (medium height, medium weight), III large--(big height, big weight) and two height/weight discordant categories: IV--so-called pyknomorphs, V--so-called leptomorphs. To assess the differences between classes the Scheffe-test was used (alpha = 0.05). It proved likewise possible to comparatively systematize length, breadth and depth measurements, circumferences and body composition characteristics in all six age groups (7-18 years) of ordinary schoolgirls and in 13-16-year-old volleyballers as in their case the average age did not differ significantly between the classes. PMID- 15830590 TI - Physical exercise, body mass index, subcutaneous adiposity and body composition among Bengalee boys aged 10-17 years of Kolkata, India. AB - A comparative study of 215 sedentary (no regular physical exercise undertaken) and 313 physically active (regular physical exercise undertaken) Bengalee boys aged 10-17 years was undertaken to investigate the differences in overall adiposity (body mass index), subcutaneous adiposity (skinfolds) and body composition (percent body fat, fat mass and fat mass index). Both groups had a similar age. The results revealed that boys who did not undertake regular physical exercise (NPE) had a significantly greater mean body mass index (BMI) compared with those who undertook regular physical exercise (PE); p < 0.001. The means for all the skinfolds as well as percent body fat (PBF), fat mass (FM) and fat mass index (FMI) were significantly higher among the NPE group. The percentile distributions of all these variables and indices were consistently higher among the NPE group. The results of ANOVA of physical exercise (PE = yes, NPE = no) and PBF, FM and FMI, with age as covariate, revealed that PE had a significant negative effect on all these measures of body composition even after controlling for the impact of age. The means in each case were greater among the NPE group. In conclusion, this study provided evidence that Bengalee boys, who undertook regular physical exercise, had significantly less adiposity compared with those who did not undertake regular physical exercise. PMID- 15830591 TI - [Postmortem injury--an impressive postmortem phenomenon that can lead to diagnostic errors]. AB - A single skull from a 6-year-old child was sent to the Anthropological Institute of the University of Zurich. It bore a considerable bone lesion in the frontal area. Closer inspection revealed that it was a postmortal lesion, resulting from the skull chafing on the river bottom whilst drifting in running water. The postmortal change, the so-called "Treibverletzung" (drift injury), is characteristic for bodies that are recovered from rivers or stillwater. A recent case from the Medical Examiners' Office from the same geographic area as the child, showed the same features of the "Treibverletzung". Since it is known from the recent case where it was found, it becomes clear that even small rivers can cause characteristic postmortal injuries. PMID- 15830592 TI - [Osteoporosis, a world problem in public health. Role of health professionals]. AB - Osteoporosis is responsible of more than two millions of femoral fractures each year in the world. This pathology concerns essentially the post-menopausal women and is frequently undetected because it is painless in the beginning. The early detection is possible by osteodensitometry eventuelly associated with biochemical markers. The prevention consists in a rich calcium-D vitamin diet, the estrogen treatment being much discussed actually. Efficient treatments (SERMs, bisphosphonates, teriparatide...) allow the reduction of fractures risks. Health professionals (physicians, pharmacists, biologists...) must give valuable information to patients on this disease, they also must recommend an early detection and an effective treatment of osteoporosis requiring a good observance. PMID- 15830593 TI - [Anaphylaxis and food allergy in children: a major problem in public health]. AB - Atopic diseases represent the most prevalent chronic illness in childhood, affecting between 15-30 % of all children in Western countries (4th problem of public health for WOH). Their incidence increased dramatically during the past 30 years. The observed increase in atopic disorders was supposed to be a consequence of changes in lifestyle factors, including improvement in public health, reduction of childhood infection, vaccination programs and changes in family size and living conditions: these factors all reflect reduced microbial burden due to increased hygiene. Food allergy and anaphylaxis in the child are also becoming main manifestations of atopy in infancy and early childhood. Five allergens are responsible for about 85 % of all cases, that can be frequently life-threatening. Diagnosis, treatment and prevention of the disease are analysed. Research on induction of tolerance and prevention were also discussed. PMID- 15830594 TI - [Academic eulogy for professor Alphonse Lafontaine]. PMID- 15830595 TI - [Cerebral mechanisms of hypnosis]. AB - The neural mechanisms underlying hypnosis and especially the modulation of pain perception by hypnosis remain obscure. Using PET we first described the distribution of regional cerebral blood flow during the hypnotic state. Hypnosis relied on revivification of pleasant autobiographical memories and was compared to imaging autobiographical material in <>. The hypnotic state was related to the activation of a widespread set of cortical areas involving occipital, parietal, precentral, premotor, and ventrolateral prefrontal and anterior cingulate cortices. This pattern of activation shares some similarities with mental imagery, from which it mainly differs by the relative deactivation of precuneus. Second, we looked at the anti-nociceptive effects of hypnosis. Compared to the resting state, hypnosis reduced pain perception by approximately 50 %. The hypnosis-induced reduction of affective and sensory responses to noxious thermal stimulation were modulated by the activity in the midcingulate cortex (area 24a'). Finally, we assessed changes in cerebral functional connectivity related to hypnosis. Compared normal alertness (i.e., rest and mental imagery), the hypnotic state, significantly enhanced the functional to modulation between midcingulate cortex and a large neural network involved in sensory, affective, cognitive and behavioral aspects of nociception. These findings show that not only pharmacological but also psychological strategies for pain control can modulate the cerebral network involved in noxious perception. PMID- 15830596 TI - Concomitant leg injuries in raptors with wing damage. PMID- 15830597 TI - Veterinary education in South Africa: the class of 1926. AB - Both Sir Arnold Theiler, appointed as Dean in 1920 when the Onderstepoort Veterinary Faculty was established, and Dr P J du Toit, who succeeded him in 1927, appear in the compound photograph of the class of 1926, constituting the 3rd group of graduates. The probable reason for having 2 deans on the photograph, which was obviously composed from shots taken individually of the outgoing and incoming deans of the faculty as well as of each of the 7 graduates, is initially analysed in this paper. This is followed by short descriptions of the life histories of the graduates. The class of 1926 was probably unique in that 2 of its members, who were employees of the Onderstepoort Research Institute, were granted special permission to study veterinary science at the Faculty. After graduating, most of the class members spent the greater part of their careers as civil servants at Onderstepoort as researchers and part-time teachers in the Faculty, as was customary for many years, or as state veterinarians in the field. Private practice did not feature strongly with them. Noteworthy is that the class produced 3 outstanding scientists, i.e. a world class parasitologist, a very eminent pathologist and Onderstepoort's 1st poultry specialist. PMID- 15830598 TI - The inheritance of liability to epistaxis in the southern African Thoroughbred. AB - This study was carried out to estimate the heritability of liability to epistaxis in the southern African Thoroughbred population. Data of all horses that suffered epistaxis while racing in southern Africa and Mauritius from 1986 to 2002 and involving 1252 bleeders were analysed. Pedigree data covering the period 1960 1986 was used as required to calculate the incidence of bleeding amongst ancestors of the post-1986 era. Only pedigrees of horses that raced were included in this study as it was not possible to predict whether non-runners would have bled had they raced. Consequently all non-runners and also those that raced overseas in countries where bleeding occurrence was not recorded, were excluded. The heritability of liability method as described by Falconer (1989) was used to estimate the relative importance of heredity and environment. For the period investigated, the population incidence for epistaxis in southern African horses was 2.1%. The estimation of heritability of liability showed that 1st-degree relatives had a figure of 55.4%. Second-degree relatives had a heritability of 41.3% and 3rd-degree relatives came in at 30.4%. The heritability of liability shown in this study could be regarded as being at the lower end of the range but could be appreciably higher. The data depict horses that bled almost exclusively on race days, as only a small percentage (approximately 5%) was reported as having bled during exercise. Accordingly, the full extent of epistaxis amongst racing Thoroughbreds in southern Africa is difficult to gauge. It is clear that epistaxis in the racing Thoroughbred has a strong genetic basis. It is suggested that horses showing frankbleeding from the nostrils after racing or exercise be suspended, and not used for breeding purposes. This should result in relatively fast progress being made towards eradicating this costly scourge of the modern Thoroughbred racehorse. PMID- 15830599 TI - Diagnostic value of magnetic resonance imaging and computed tomography for oral masses in dogs. AB - The purpose of this study was to determine the diagnostic value of magnetic resonance imaging (MRI) and computed tomography (CT) in oral masses of dogs. Nineteen dogs underwent clinical, MR and CT examinations. Eleven malignant and ten non-malignant masses were evaluated. Osteosarcoma was the most commonly found malignant oral mass and gingival hyperplasia was the most commonly found benign mass. The results showed that MRI provided more accurate information regarding the size of the masses and invasion of adjacent structures although MRI and CT show similar accuracy in assessment of bone invasion. Calcification and cortical bone erosion was better seen on CT images. Whereas contrast-MRI provided useful additional information, contrast-CT had no added benefit. In general, oral masses located in the caudal mandible, oropharynx and maxilla are better evaluated using MRI, once the histological type has been verified. PMID- 15830600 TI - Retrospective study of snake envenomation in 155 dogs from the Onderstepoort area of South Africa. AB - A retrospective study was undertaken to evaluate the incidence, signalment, haematological and biochemical changes, therapy, and outcome of dogs presented to the Outpatients section of the Onderstepoort Veterinary Academic Hospital for confirmed snake envenomation. Three hundred and seventy-six records of dogs presented for snake envenomation from 1998 to 2002 were reviewed and 155 were selected on the basis of there being a positively identified snake. The 2 most commonly encountered snake envenomations in dogs were puff-adders (Bitis arietans) and snouted cobras (Naja annulifera annulifera). The majority of cases (56%) occurred in the autumn (March to May), with most being bitten by puff adders. Dogs were 3 to 168 months old with a median of 36 months. No sex predilection was identified. Ten per cent of cases died because of the snake envenomation. Fifty-seven per cent and 43% of snakebites were puff-adders and cobras, respectively. There was no difference in mortality between the 2 groups of snakes. Of the cobras 60% were the snouted cobra, 14% Mozambique spitting cobra, and 24% rhinkals. Swelling in the area of the bite, usually the face and forequarters, was the primary clinical abnormality. Significant haematological findings were leukocytosis (median 17.3 x 10(9)/l; range 0.4-44), neutrophilia (median 13.6 x 10(9)/l; range 0.3-39.9), band neutrophilia (median 0.4 x 10(9)/l; range 0-5.32), and thrombocytopaenia (median 124 x 10(9)/l; range 3-555). Dogs envenomated by a puff-adder and Mozambique spitting cobra had a greater degree of thrombocytopaenia: median of 68 and 66, respectively, versus 243 for the cobra group. The most commonly used treatments were intravenous fluids, antibiotics and glucocorticoids. Thirty-eight dogs were treated with polyvalent antiserum: 9 for puff-adder envenomation and 29 for cobra envenomation. Only 2 of the dogs that received antisera died, both of them of cobra envenomation. The study concluded that snake envenomation in dogs is associated with high morbidity but moderate mortality rate and that the most significant haematological abnormality is thrombocytopaenia. PMID- 15830601 TI - Knowledge and attitudes of cattle owners regarding trypanosomosis control in tsetse-infested areas of Uganda. AB - A pilot survey using a structured questionnaire was conducted in Tororo and Busia districts of Uganda on the knowledge and attitudes of cattle owners regarding tsetse fly and trypanosomosis control, in order to understand factors that hindered their full participation. A total of 81 cattle owners was randomly selected and interviewed, of which 92.5% were aware of tsetse flies and trypanosomosis and 87.6% recognised animal trypanosomosis as a problem in the area. Most cattle owners were aware of tsetse fly trapping (76.5%), isometamidium chloride use (55.5%), diminazene aceturate use (48%) and pour-on applications (18.5%). However, knowledge did not coincide with the application of control measures. Despite the widespread awareness, tsetse fly trapping and pour-on applications were used by only a small percentage of cattle owners (7.5% applied tsetse fly trapping while 76.5% were aware of it; 1.2% applied pour-on insecticides while 18.5% were aware of them). Differences between awareness and application were highly significant for tsetse fly trapping (chi2 = 67.8, d.f. = 1, P < 0.001) and pour-on applications (chi2 = 10.8, d.f. = 1, P < 0.05), but not for isometamidium chloride use (chi2 = 0.08, d.f. = 1, P = 0.77) and diminazene aceturate use (chi2 = 0.00, d.f. = 1, P = 1.00). Most cattle owners (97.5%) were willing to participate in future control programmes, but preferred participating on a group basis (85.2%) rather than individually (14.8%). The 4 most favoured control options in order of importance were: fly traps supplied by the government and maintained by cattle owners; contribution of labour by cattle owners for trap deployment; self-financing of trypanocidal drugs and self-financing of pour-on insecticide. The control options that should be selected in order to elicit full participation by cattle owners are discussed. PMID- 15830602 TI - Aldicarb poisoning of dogs and cats in Gauteng during 2003. AB - A survey of aldicarb poisoning in companion animals was conducted by posting questionnaires to all private practitioners in Gauteng Province, South Africa. The survey was designed to determine the percentage of aldicarb cases seen, treatment regimen, clinical signs observed, proposals for preventative actions and more effective treatments. Other questions included duration of treatment, survival rate, cost to clients, post mortem findings and reasons for poisonings. PMID- 15830603 TI - Evaluation of hydroxypropyl-beta-cyclodextrin in the treatment of aldicarb poisoning in rats. AB - Cyclodextrins are ring-shaped oligosaccharides with a hydrophilic exterior and a hydrophobic interior. The interior cavity is capable of complexing fat-soluble molecules small enough to fit inside. Sprague-Dawley rats were used to evaluate the efficacy of hydroxypropyl-beta-cyclodextrin as treatment of aldicarb poisoning in rats. Survival times in the majority of rats dosed with aldicarb and receiving intravenous cyclodextrin were longer compared with the control rats only dosed with aldicarb per os. PMID- 15830604 TI - Susceptibility of 7 freshwater gastropod species in Zimbabwe to infection with Gastrodiscus aegyptiacus (Cobbold, 1876) Looss, 1896. AB - Gastrodiscosis outbreaks due to Gastrodiscus aegyptiacus were recorded in horses in the vicinity of Harare, Zimbabwe, in the absence of Bulinus forskalii, B. senegalensis and Cleopatra sp. which are considered to be the only intermediate host snails. This suggested the possibility of other snail species acting as intermediate hosts in the life cycle of the trematode. A study was carried out to determine the susceptibility of 7 freshwater snail species to infection with G. aegyptiacus. First generation (F-1) of 5 freshwater pulmonate snail species, Bulinus tropicus, Bulinus globosus, Biomphalaria pfeifferi, Helisoma dyuri and Physa acuta that were bred in the laboratory, and 2 prosobranch snail species, Melanoides tuberculata and Cleopatra sp. that were collected from the field were used in this study. Data pertaining to mortalities and cercariae shedding were recorded throughout the experimental period. The prosobranch snails, M. tuberculata and Cleopatra sp. were susceptible to G. aegyptiacus with a minimum prepatent period of 45 days and 54 days, respectively. Bulinus tropicus, P. acuta and H. duryi were susceptible as evidenced by the presence of different generations of rediae and mature cercariae on dissection at 59 days post infection although attempts to induce the snails to shed from 28 days post infection did not produce cercariae. Bulinus globosus and Bio. pfeifferi were refractory to infection. The results revealed the ability of G. aegyptiacus to infect M. tuberculata, Cleopatara sp., B. tropicus, P. acuta and H. duryi under experimental conditions and this may explain the recorded outbreaks of gastrodiscosis in equine populations in Zimbabwe in the absence of the known intermediate hosts. Bulinus tropicus is considered as the most likely major intermediate host of G. aegyptiacus because of its wide distribution in Zimbabwe and is well adapted to a wide variety of environments. PMID- 15830605 TI - Effect of strategic deworming of village cattle in Uganda with moxidectin pour-on on faecal egg count and pasture larval counts. AB - Strategic application of moxidectin pour-on (Cydectin) was evaluated in Uganda for its effect on pasture larval counts and gastrointestinal nematode faecal egg counts in village cattle kept under tethering (semi-intensive) grazing management. The strategic deworming schedule involved treating cattle twice at an interval of 2 months, at the end of the 1st wet season and during the 2nd wet season. Two groups of 30 cattle, each consisting of a treated and a control group, were examined for nematode infections every 4 weeks from June 1999 to January 2000. The treated group had significantly lower mean faecal egg counts than the untreated groups (t-value = 2.47, P < 0.05). Generally, the pasture larval counts on treated farms were lower than on untreated ones, but not significantly so (t-value = 2.22, P = 0.068). Pasture larval counts with different nematode species on treated farms were lower than on untreated ones, but the differences were not significant for Haemonchus spp. (t-value = 1.68, P = 0.145), Oesophagostomum spp. (t-value = 1.87, P = 0.111), Trichostrongylus spp. (t-value = 1.93, P = 0.102), Dictyocaulus spp. (t-value = -0.74, P = 0.485) and Cooperia spp. (t-value = -1.00, P = 0.356). Treated farms did, however, have significantly lower pasture larval counts of Bunostomum spp. (t-value = 4.64, P < 0.05). This study has revealed that the application of moxidectin pour-on on cattle has an effect on faecal egg count and pasture contamination under the tethering grazing system. Moxidectin pour-on and the strategic deworming schedule evaluated here could be used for the control of gastrointestinal nematode infections in cattle by small-scale farmers who practise tethering or semi intensive grazing management in Uganda and other tropical countries, especially where there is a bimodal rainfall pattern. PMID- 15830606 TI - [Early anemia in diabetic nephropathy]. AB - The number of diabetic patients with renal disease increased significantly in the last years worldwide. Anemia is an important and frequent component of diabetic nephropathy that may begin early in the course of the chronic renal disease of diabetics, and is more severe in diabetic patients with renal disease than in non - diabetic renal patients controlled for the same level of renal function. The reason for the anemia is decreased erythropoietin level caused by diminished production and, in a lesser degree, by increased excretion of erythropoietin in the urine. There is a close connection between diabetic nephropathy, anemia and cardiovascular complications. On the basis of small studies correction of anemia may decrease the progression of diabetic nephropathy and cardiovascular complications. However, the result of ongoing large randomised controlled studies are required to get "evidence-based" data to prove that correction of anemia has beneficial effects on microvascular and macrovascular diabetic complications, particularly cardiac disease, and on progression of diabetic nephropathy. PMID- 15830607 TI - [Laboratory diagnosis of polysystemic autoimmune diseases]. AB - After a short description of the mechanism of the development of polysystemic autoimmune diseases, this paper presents the in vitro laboratory methods suitable for the characterization of the humoral and cellular elements of immune system, furthermore, summarizes the characteristic autoantibodies and other laboratory alterations in these diseases. PMID- 15830608 TI - [Incidence and pathologic distribution of esophageal cancers at the gastro esophageal junction between 1993-2003]. AB - There was a significant change in the histology of oesophageal cancers in the last few decades. The incidence of oesophageal adenocarcinoma has risen considerably, now it is equally or even more prevalent than squamous cell cancers in some North American and Western European countries. As no Hungarian data is available, the authors' aim was to investigate the prevalence and histology of oesophageal and gastrooesophageal junction cancers in the last decade. PATIENTS AND METHODS: 451 patients diagnosed with oesophageal (n = 371, 296 male/75 female, mean age at diagnosis: 57.9 SD 10.1 years) or cardia (n = 80, 58 male/22 female, mean age at diagnosis: 65.2 SD 13.4 years) cancer between 1st of January 1993 and 31st of December 2003 at the 1st Internal Medicine and 1st Surgery Department of Semmelweis University were enrolled. Pathology and clinical data were analysed retrospectively. RESULTS: 93% (n = 345) of the patients with oesophageal cancer had squamous cell carcinomas, while adenocarcinoma was only diagnosed in 15 (4%) patients. Mean age at diagnosis was lower in patients with squamous cell cancer (57.4 SD 10.0 years) compared to patients with adenocarcinoma (66.9 SD 8.8 years, p = 0.001). Male-to-female ratio was 4:1 in patients with squamous cell carcinoma (277/68) and undifferentiated carcinoma (9/2), while it was 2:1 in patients with adenocarcinoma (10/5). According to the location 1.3% of cancers of the midthoracic oesophagus and 8.6% of the lower oesophagus were adenocarcinoma. The proportion of adenocarcinoma remained stable over the observed period (1993-1997: 3.7% vs. 1998-2003: 4.3%). In contrast, 71.25% (57/80) of the gastrooesophageal junction cancers and overall 15.9% (72/451) of the cancers of the oesophagus and gastrooesophageal junction were adenocarcinoma (1993-1997: 17.2% vs. 1998-2003: 14.7%). CONCLUSIONS: Since only a few percentages of authors patients with oesophageal cancers were diagnosed to have adenocarcinoma and its proportion remained stable over the observed period, it seems that in contrast to North American and Western European countries, adenocarcinoma is still infrequent in Hungary. PMID- 15830609 TI - [Presidential remarks of the Special Collegium for Surgery]. PMID- 15830610 TI - [Asymptomatic bladder cancer evaluated by gynecological sonography]. AB - In the past few years, ultrasonography has become the most widespread and easily available diagnostic procedure in gynecological practice. the high-frequency transvaginal probe provides a detailed visualization of the reproductive organs and the urinary bladder. however, in spite of this generally applied examination procedure it's rare when the transvaginal ultrasonography is used to identify bladder tumor. the urologist rather use transabdominal ultrasound for the detection of bladder tumors. the authors report a case in which gynecological sonography contributed to the screening and evaluation of an asymptomatic bladder cancer. PMID- 15830611 TI - [Treatment of genital candidiasis]. PMID- 15830612 TI - Praxis skills in young children with Down syndrome, other developmental disabilities, and typically developing children. AB - This study explored whether young children with Down syndrome show praxis deficits that impact activities of daily living, and whether these deficits are specific to Down syndrome. We compared the performance of young children with Down syndrome, a mental age-matched group of children with developmental disabilities of mixed or unknown etiologies, and a group of typically developing infants and toddlers on praxis tasks and overall adaptive behavior (Vineland Adaptive Behavior Scales). Children with Down syndrome showed poorer overall motor functioning than the developmental disabilities comparison group as measured by the Vineland Adaptive Behavior Scales, F(2, 47) = 5.24, p < .01 (using one-way analysis of variance [ANOVA]). A one-way multivariate analysis of variance also showed that children with Down syndrome performed significantly worse overall than the developmental disabilities comparison group on a battery of praxis tasks, F(7, 18) = 2.95, p < .05, and a series of object retrieval tasks, F(7, 18) = 2.95, p < .05, suggesting a deficit in praxis that is specific to Down syndrome. Children with Down syndrome elicited significantly more help than both comparison groups during object retrieval trials, F(2, 48) = 4.94, p < .01 (using one-way ANOVA). When chronological age was partialled out, a strong relationship was observed between praxis and adaptive functioning in Down syndrome, r(8) = .69, p < .05. These findings suggest that young children with Down syndrome may need targeted interventions that focus on both praxis skills and motivational orientation. PMID- 15830613 TI - The relation between length of institutionalization and sensory integration in children adopted from Eastern Europe. AB - OBJECTIVE: To examine the relation between length of institutionalization and sensory integration in children adopted from Eastern Europe. METHOD: The Sensory Integration and Praxis Tests (SIPT) and the Developmental and Sensory Processing Questionnaire were administered to 60 adopted children, 4 years to 8 years, 11 months of age. Thirty had longer institutionalization histories (mean: 34 months; LIH group) and the other 30 had shorter institutionalization histories (mean: 3 months; SIH group). RESULTS: The LIH group demonstrated significantly lower scores than the SIH group on the SIPT in vestibular-proprioceptive, visual, and praxis areas, and effect sizes ranged from .09 to 1.13. The LIH group also had significantly more frequent behaviors suggestive of sensory modulation dysfunction compared to the SIH group, particularly in touch and movement seeking, vision, and audition. Effect sizes ranged from 0 to 1.39. CONCLUSION: Longer lengths of institutionalization are associated with more atypical sensory discrimination, praxis, and sensory modulation scores in children adopted from Eastern European orphanages. The areas of sensory integration that appear to be more vulnerable to deprived conditions in early childhood are vestibular proprioceptive, tactile, visual, auditory, and praxis. Adopted children with lengthy periods of institutionalization may benefit from occupational therapy for early sensory integrative and developmental screenings. PMID- 15830614 TI - Typical children's responsivity patterns of the tactile and vestibular systems. AB - OBJECTIVES: The purposes of this study were to investigate the responsivity patterns of typical 3- and 4-year-old Israeli children to tactile or vestibular stimulation, or both, and to examine whether differences in these patterns exist between them with respect to age and gender. METHOD: The study sample consisted of one hundred seventeen 3-year-old and one hundred forty-three 4-year-old healthy Israeli children (N=260). Mothers of these children completed a comprehensive tactile and vestibular responsivity questionnaire. RESULTS: The subjects' tactile and vestibular responsivity scores were neither hyperresponsive nor hyporesponsive. In addition, neither age nor gender was found to significantly differentiate between respective participant groups for hypo- or hyperresponsive behaviors. CONCLUSION: Typical Israeli children can be characterized by moderate responsivities to tactile and vestibular stimulation. Responsivity to tactile and vestibular input was similar for 3- and 4-year-olds, across genders. PMID- 15830615 TI - A holistic approach to teaching evidence-based practice. AB - Articles pertaining to evidence-based practice have become increasingly visible in occupational therapy literature. Authors have defined and described evidence based practice for occupational therapy (Lloyd-Smith 1997; Taylor 1997), identified resources that therapists can draw upon to inform their practice (Hayes & McGrath, 1998; Lieberman & Scheer, 2002), and articulated strategies to gather research evidence in practice (Tickle-Degnen 2000a; Unsworth, 2000). As associate editor of the American Journal of Occupational Therapy Evidence-Based Practice Forum, Tickle-Degnen has also written a series of articles that explain how to engage clients in the research process, and communicate research evidence to clients and families (1998, 2002). PMID- 15830616 TI - Genetics in occupational therapy education: a survey of professional entry-level programs. AB - PURPOSE: Advances in genetics indicate a need for occupational therapists to develop literacy and skills in genetics as it relates to lifestyle and occupation. The purpose of this study is to identify genetics content areas taught, instructional methods used, and the importance of teaching genetics at the entry-level in occupational therapy curricula. METHOD: A questionnaire was sent to all entry-level occupational therapy educational programs (N=157). Structured mailing and follow-up were used. RESULTS: The response rate was 63.9%. Most respondents (47%) rate teaching genetics as "moderately important." Genetics content is predominately taught at the introductory or knowledge level rather than at the integration and application level. Respondents indicate a lack of time and space for genetics content and of faculty interest and expertise. DISCUSSION: As occupational therapy practice evolves to include new genetics, curriculum change will need to be implemented. Development of teaching materials and methods addressing genetics is recommended. PMID- 15830617 TI - The role of the school-based occupational therapist in secondary education transition planning: a pilot survey study. AB - OBJECTIVE: The purpose of this study was to investigate school-based occupational therapists' knowledge of transition planning, their degree of participation in assessment and intervention of students requiring transition services, and to identify potential barriers limiting therapists' participation in transition services. METHOD: Using survey methods, a questionnaire was mailed to a random sample of therapists listed as members of the School System Special Interest Section of the American Occupational Therapy Association. Eighty therapists from all geographical regions within the continental United States and who identified themselves as working with students 13-21 years of age in an educational setting, participated in the study. The response rate was 20%. RESULTS: The majority of participants reported that they understood the terminology associated with the Individuals with Disabilities Education Act (IDEA) of 1990 and the 1997 IDEA amendments definitions of transition planning at the secondary level, but were less likely to apply that knowledge to the transition planning process. The majority of therapists reported minimal participation in secondary education transition planning assessment and intervention for students with disabilities. Most respondents believed that they were not contributing to the transition planning process in a manner that maximized their skills, and identified several barriers that they believed hindered greater participation. CONCLUSION: This pilot study suggests that occupational therapists may not be participating in transition services to their fullest potential. While the low response rate in this study precludes generalization, this information is important to guide further study as well as to shape efforts to increase occupational therapy's role in this important service area within school-based practice. PMID- 15830619 TI - The meaning of computers to a group of men who are homeless. AB - The purpose of this pilot study was to explore the experience with computers and the meaning of computers to a group of homeless men living in a long-term shelter. This descriptive exploratory study used semistructured interviews with seven men who had been given access to computers and had participated in individually tailored occupation based interventions through a Work Readiness Program. Three themes emerged from analyzing the interviews: access to computers, computers as a bridge to life-skill development, and changed self-perceptions as a result of connecting to technology. Because they lacked computer knowledge and feared failure, the majority of study participants had not sought out computers available through public access. The need for access to computers, the potential use of computers as a medium for intervention, and the meaning of computers to these men who represent the digital divide are described in this study. PMID- 15830618 TI - An outcome in need of clarity: building a predictive model of subjective quality of life for persons with severe mental illness living in the community. AB - PURPOSE: The study purpose was to construct a predictive model of subjective quality of life for persons with severe mental illness living in the community with particular attention to participation in occupations. METHOD: Persons with severe mental illness (N=154) rated their subjective quality of life. Several measures for each of the following categories of variables were completed: demographics, clinical, social participation, and self-measured well-being. Regression analysis was used to determine the significant predictors for each category and then to build the predictive model from these significant variables. RESULTS: Symptom distress accounted for the most variance (33%) in subjective quality of life, followed by psychological integration (3%) and physical integration (2%). CONCLUSIONS: The study suggests that occupational therapists should attend to subjective experience of symptoms to influence quality of life. Therapists are also in a good position to address their clients' sense of belonging to their communities and to enable community participation. PMID- 15830620 TI - Finding common ground with patients: the centrality of compatibility. AB - This research examines the lived experiences of six novice occupational therapists (0-2 years experience) and eight career occupational therapists (> or = 5 years in the everyday practice in adult rehabilitation. The study focused specifically on therapists' experiences of working together with patients and the ways in which these experiences unfolded over the course of therapy. Data were generated using a combination of phenomenological interviews with all 14 therapist-participants (Phase I) and participant observation of four therapist patient dyads throughout the course of rehabilitation therapy (Phase II). Verbatim transcriptions of all interviews from Phase I and Phase II and field notes from Phase II were analyzed using a narrative approach. The analysis revealed that Finding Common Ground was a shared meaning of working together with patients regarding therapy goals and expectations. Findings suggest that occupational therapists may resist negotiating differences with patients over therapy goals and expectations and instead rely primarily on compatibility as the basis of finding common ground. Findings reveal the importance for occupational therapists to be (1) open to negotiating differences with patients over goals and expectations, (2) aware of the ideologies that may influence their practices, and (3) adequately prepared to deal effectively with the challenging interpersonal aspects of practice. PMID- 15830622 TI - Waiting to get better: a dilemma regarding habits in daily occupations after stroke. AB - The purpose of this study was to explore and gain an understanding of how habits are experienced when performing daily occupations after a stroke. In-depth interviews were conducted with 7 participants and a constant comparative method was used for data analysis. Four men and three women with stroke between the ages of 42 and 82 years participated in the study. The interviews were conducted 1.5 to 7 months after the participants had been discharged from hospital to their home. The findings show that the participants experienced frustration when performing daily occupations because changes in their performance meant that former habits could not automatically be reestablished; daily occupations had to be re-organized and planned with greater deliberation than had been required prior to the stroke. In reestablishing their daily occupations, the participants experienced an ongoing conflict about whether or not to develop new habits. Although adaptation and change would be beneficial in the short-term, both also represented giving up possible improvements, and participants seemed to consider that their eventual recovery and independence would be compromised if they allowed themselves to alter their habits. This dilemma led to a sensation of waiting: waiting to get better, waiting for another solution and waiting for the treatment to make an impact. As a result, few new habits were established in daily occupations. The findings suggest that occupational therapists need to be aware of the dilemma clients may perceive regarding decisions about whether or not to adapt and develop new habits during poststroke recovery. PMID- 15830621 TI - Professional strategies in work-related practice: an exploration of occupational and physical therapy roles and approaches. AB - Both occupational and physical therapy have historical roots in worker rehabilitation; however, the core philosophies and areas of expertise of these professions suggest there may be inherent differences in the ways each approaches therapeutic intervention. This study surveyed 600 occupational and physical therapists in the United States to determine the nature and scope of work-related practice, and the degree to which occupation-based strategies are used by either profession. The overall response rate was 54% (n = 324), and 76% of respondents (n = 246) were actively engaged in providing work-related services. Results indicate that both professions provide all services commonly associated with work related therapy in almost equal proportions. Few significant differences were identified between the professions in terms of approaches to therapy, and findings did not reveal the use of occupation-based interventions to any greater degree by occupational therapists. PMID- 15830623 TI - Expanding the community role for occupational therapy: becoming political in the corporate sector. PMID- 15830624 TI - Scholarship and practice: bridging the divide. AB - There is evidence that the growth theory and research about occupation produced by academics are not consistently translated into occupational therapy practice. Rather, practitioners often report finding such theory and research to be of limited relevance to and/or difficult to implement in their everyday work. Although many factors contribute to this academic-practice gap one important contributor is how knowledge is traditionally viewed and generated in academia. Changes in traditional views about academic knowledge and its relationship to practice both outside and within occupational therapy are discussed. Ways that occupational therapy scholarship can be more clearly grounded in everyday practice are examined drawing from experience with participatory research. Consideration is given to how researchers and practitioners might share power and have a more effective dialogue. PMID- 15830625 TI - Quick turnover of physician groups raises red flags for ED managers. AB - With proper planning and awareness, impact of contract changes can be minimized. Make sure goals and objectives are understood clearly by you and your physicians' group. Recognize transitions always are stressful; be on guard for staff morale problems. Avoid midwinter staff turnover, and give new staff at least six months' advance notice before their move. PMID- 15830626 TI - Physician-owned group or corporation: pros, cons. PMID- 15830627 TI - ED screening changes put pressure on competitors. AB - Counseling patients about other options for nonurgent care can help ease the pressure on your ED. This process has the potential to decompress your ED by as much as 40%. Implementing post-screening counseling will put pressure on your competitors to follow suit. If the patient does not have an emergency condition, seek confirmation of payer source before treatment. PMID- 15830628 TI - Use of e-mail raises many HIPAA concerns for EDs. AB - Learn which types of electronic communications are most secure and who is permitted to see them to avoid regulatory violations. Use of common commercial e mail services will not offer the necessary protection required by the Health Insurance Portability and Accountability Act of 1996. ED managers should be familiar with guidelines, especially electronically protected health information. Follow-up e-mails to primary care providers are only appropriate if your line and the recipient's line are encrypted. PMID- 15830629 TI - An increase in obesity takes a toll on EDs. AB - Make sure you have the necessary equipment, and train your staff to treat obese patients with respect. Consider the acquisition and addition of oversized beds, wheelchairs, and blood pressure cuffs. Renovations of doorways and commodes may be necessary to accommodate larger patients. Sensitivity training can help improve satisfaction levels among obese patients. PMID- 15830630 TI - Does EMTALA apply to patients who go to ED for a scheduled appointment? PMID- 15830631 TI - Gingival overgrowth in children: epidemiology, pathogenesis, and complications. A literature review. AB - Gingival overgrowth is the enlargement of the attached gingiva due to an increased number of cells. The most prevalent types of gingival overgrowth in children are drug-induced gingival overgrowth, hereditary gingival fibromatosis (HGF), and neurofibromatosis I (von Recklinghausen disease). Gingival overgrowth induced by drugs such as phenytoin, nifedipine, and cyclosporin develops due to an increase in the connective tissue extracellular matrix. According to epidemiologic studies, it is more prevalent in male children and adolescents. There is an additive effect of those drugs on the degree of gingival overgrowth. Genetic heterogeneity seems to play an important role in the development of the disease. Functional difficulties, disfigurement, increased caries, and delayed eruption of permanent teeth are the main complications of drug-induced gingival overgrowth. HGF is the most common syndromic gingival enlargement in children. This autosomal dominant disease usually appears at the time of eruption of permanent dentition. Histologically, it is characterized by highly collagenized connective tissue. The most important complications are drifting of teeth, prolonged retention of primary dentition, diastemata, and poor plaque control. Neurofibromatosis I is an autosomal dominant disease more common in mentally handicapped individuals. Gingival overgrowth is caused by the formation of plexiform neurofibromas in the connective tissue of the gingiva. Plexiform neurofibromas are pathognomonic of the disease and consist of hypertrophic nerves arranged as lobules in the connective tissue. Complications of the disease are multiple and severe due to neurofibromas and their occasional malignant transformation. PMID- 15830632 TI - Longitudinal study of periodontal disease and edentulism with rates of bone loss in older women. AB - BACKGROUND: Previous cross-sectional studies have suggested a link between periodontal disease and osteoporosis. The purpose of the present study is to evaluate the association between changes in bone mineral density (BMD) and clinical signs of periodontal tissue destruction and tooth loss over a 2-year period. METHODS: A total of 398 women (mean age 75.5 years) were randomly selected for an ancillary study of periodontal disease; osteoporosis in association with the presence or absence of teeth was evaluated. Osteoporosis in association with periodontal disease was also evaluated. All subjects were participants at the Pittsburgh Clinical Center for the Study of Osteoporotic Fractures (SOF), a prospective cohort study of women 65 years of age or older designed to determine risk factors for fractures. Oral health examinations, including periodontal probing and attachment loss, were performed at the fourth clinical visit, an average of 6 years after baseline. BMD of the total hip and its subregions was measured using dual energy x-ray absorptiometry at the time of dental examination and 2 years later. Results are expressed as annual percentage change. RESULTS: A total of 145 (36.4%) women were edentulous and 163 (80.7%) of the dentate women (N = 253) had periodontal disease. Dentate women reported higher education (P <0.001) and a higher calcium intake (P= 0.002). Absolute BMD and percentage change in BMD were similar in dentate and edentulous women. We found no difference in BMD or in absolute or percentage change in BMD between women with or without periodontal disease. CONCLUSION: Little evidence exists for an association between edentulousness, periodontal disease, and longitudinal changes in BMD. PMID- 15830633 TI - Reconstitution of a hyperinflammatory prostaglandin E2 response to Porphyromonas gingivalis challenge in severe combined immunodeficient mice. AB - BACKGROUND: The purpose of this study was to determine whether lymphocytic cells regulate the monocytic hyperinflammatory trait (MO+) in chronic periodontitis patients. Using a P. gingivalis challenge model in severe combined immunodeficient (SCID) mice, we tested the effects of adoptively transferred human peripheral blood leukocytes from gingivitis and chronic periodontitis diabetic and non-diabetic individuals on monocytic responses. METHODS: This response was examined using the subcutaneous tissue chamber infection model. Three weeks following cell reconstitution, all SCID mice were challenged with 10(9) colony forming units of live P. gingivalis HG405. Chamber contents were collected at day 7 after bacterial challenge for prostaglandin E2 (PGE2) analysis and chamber rejection monitored up to day 30. Gingival crevicular fluid (GCF) samples were collected from all patients for PGE2 analysis. Both chamber fluid- and GCF-PGE2 levels were determined by enzyme-linked immunosorbent assays (ELISA). RESULTS: Significantly elevated GCF-PGE2 levels were found in diabetic as well as in non-diabetic patients with moderate/advanced periodontitis compared to diabetic and non-diabetic subjects with gingivitis/mild periodontitis at P= 0.01 and P= 0.001, respectively. As reflected in chamber fluid PGE2 levels and percentage chamber rejection, lymphocytic sensitization to P. gingivalis occurred in both diabetics and non-diabetics with moderate/advanced periodontitis, but not in diabetics and non-diabetics with gingivitis/mild periodontitis. CONCLUSIONS: These results suggest that the exaggerated monocytic inflammatory response trait (MO+) associated with moderate/advanced chronic periodontitis is due, at least in part, to lymphocytic modulation, while the directional findings for lymphocytes from diabetic subjects deserve further investigation. Our findings further demonstrate that the SCID mouse model is a useful animal model to study human immune responses to periodontal microorganisms. PMID- 15830634 TI - The effects of a bioabsorbable barrier membrane containing safflower seed extracts on periodontal healing of 1-wall intrabony defects in beagle dogs. AB - BACKGROUND: Recently, there has been much research done into the regenerative potential of materials used in oriental medicine. In several studies, evidence was found that these materials have an effect on bone regeneration. Among these materials, safflower seeds are of particular interest as they have been used for the treatment of blood stasis, bone fracture, and osteoporosis in traditional Korean medicine. In addition, they are known to have anti-inflammatory effects. The objective of this study is to evaluate the periodontal tissue regenerative effects of a bioabsorbable barrier membrane (polylactide glycolic acid electro spun non-woven membrane) containing safflower seed extracts applied to surgically created 1-wall intrabony defects in beagle dogs. METHODS: One-wall intrabony defects were surgically created bilaterally at the mesial and distal sides of the mandibular second premolars and mesial side of the fourth premolars. These defects were randomly assigned either to the surgical control group which received a flap operation only or to one of two experimental groups consisting of defects which received a guided tissue regenerative procedure with either a bioabsorbable membrane (PLGA) or a bioabsorbable membrane containing safflower seed extracts (SSE/PLGA). The dogs were sacrificed 8 weeks after the operation, and a comparative histological examination was done. RESULTS: The new cementum formation was 2.49+/-0.41 mm in the surgical control group, 3.22+/-0.35 mm in the PLGA group, and 3.67+/-0.82 mm in the SSE/PLGA group. The extent of new cementum formation in barrier groups was significantly different from the surgical control group (P <0.05). The amount of intrabony cementum was 1.75+/-0.06 mm, 2.40+/-0.33 mm, and 2.70+/-0.81 mm for the surgical control group, the PLGA group, and the SSE/PLGA group, respectively; the amount of infrabony cementum in the barrier groups was significantly different from the surgical control group (P<0.05). The value of the suprabony cementum was 0.73+/-0.48 mm, 0.82+/-0.21 mm, and 0.97+/ 0.09 mm for the surgical control group, the PLGA group, and the SSE/PLGA group, respectively, with no significant differences being observed among the treatments. The amount of new alveolar bone formation was 1.74+/-0.25 mm, 2.36+/ 0.30 mm, and 2.64+/-0.74 mm for the surgical control group, the PLGA group, and the SSE/PLGA group, respectively, with a significant difference exhibited between the surgical control group and other groups (P <0.05). Superficial root resorption was often observed, but ankylosis was not present. CONCLUSION: Our results suggest that surgical application of polylactide glycolic acid non-woven membrane with or without safflower seed extract could promote the regeneration of alveolar bone and cementum in intrabony periodontal defects. PMID- 15830635 TI - Human gingival tissue reactions to silk and expanded polytetrafluoroethylene sutures. AB - BACKGROUND: Wound closure by suturing is a critical and significant event in general as well as in oral surgery. Information regarding tissue reactions to different suture materials appears incomplete and inconsistent, particularly in humans. Thus, the purpose of the present study was to evaluate clinically and histologically tissue reactions to silk and expanded polytetrafluoroethylene (ePTFE) suture materials placed in human oral tissues. METHODS: Twelve patients undergoing periodontal treatment, who had bilateral periodontal defects in the maxillary premolar and molar region scheduled for resective surgery, were included. In each patient, one single interrupted ePTFE suture (CV-5) and one single interrupted silk suture (4-0) were inserted in gingival tissues in contralateral jaw quadrants 10 days prior to scheduled surgery and a second set of sutures was inserted at 7 days prior to surgery. After placement, the length of the suture embedded in the tissue as well as the slack of the suture loop were measured and tabulated. The measurements were repeated on the day of surgery. At this time biopsy specimens, including the suture loop and surrounding tissue, were harvested and processed for histologic analysis. The inflammatory response was evaluated by measuring the thickness of the perisutural epithelium and the diameter of the connective tissue infiltrate, by estimating the proportion of inflammatory cells to epithelial cells, and by recording the presence or absence of bacterial plaque along the suture track. RESULTS: At 7 days, silk sutures showed a significantly higher degree of slack of the suture loop than did ePTFE sutures (P= 0.02), while the difference was not significant at 10 days (P= 0.21). The mean change of "tissue bite" was greater for silk compared to ePTFE at 7 as well as at 10 days. However, none of the differences were statistically significant (P= 0.43). The thickness of the perisutural epithelium increased significantly from 7 to 10 days for both suture materials, while the difference in proportion of inflammatory cells to epithelial cells was not significant (P= 0.43). An overall significantly greater diameter of connective tissue infiltrate was present around the silk sutures. Bacterial plaque was detected in 10 of 11 silk and four of 11 ePTFE suture channels at 7 days, and eight of 10 and four of 11 suture channels at 10 days. CONCLUSIONS: The results revealed that placement of sutures in gingival tissues elicits an inflammatory reaction and that the magnitude of this reaction may vary with the suture material used. Braided silk sutures apparently cause a more extensive inflammatory tissue reaction in an environment characterized by moisture and infectious potential. PMID- 15830636 TI - Initial apical migration of junctional epithelium in rats following application of lipopolysaccharide and proteases. AB - BACKGROUND: Apical migration of junctional epithelium (JE) occurs in association with periodontal pocket formation. The aim of this study was to investigate the gingival changes occurring during apical migration of the JE following application of factors associated with inflammatory periodontal disease pathogenesis. METHODS: Six-week-old male Wistar rats were divided into six groups: three experimental groups to investigate gingival changes following 2, 4, and 8 weeks topical application of lipopolysaccharide (LPS) and proteases and three control groups using pyrogen-free water. After 2, 4 or 8 weeks, nuclear DNA fragmentation was detected in periodontal ligament (PDL) fibroblasts using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method, and proliferative activities of the basal cells and fibroblasts were evaluated through expression of proliferating cell nuclear antigen (PCNA). Collagen destruction was examined histologically. RESULTS: Gingiva treated with LPS and proteases showed an increase in PCNA-positive basal cells but not the fibroblasts. Collagen destruction was observed at 2 weeks; apical migration of the JE and TUNEL-positive fibroblasts was seen at 4 weeks. CONCLUSIONS: Following application of LPS and proteases to rat gingival sulci, the apical migration of the JE appears to occur simultaneously with the apoptosis of PDL fibroblasts, which in turn follows proliferation of the basal cells and collagen destruction. PMID- 15830637 TI - Modulation of clinical expression of plaque-induced gingivitis: interleukin-1 gene cluster polymorphisms. AB - BACKGROUND: The purpose of the present study was to determine the association of interleukin-1 (IL-1) gene polymorphisms with clinical parameters of gingivitis in a large experimental gingivitis trial and with each of two subgroups, high responders (HR) and low responders (LR), with distinct susceptibility to gingivitis. METHODS: Ninety-six systemically and periodontally healthy non smokers, 46 males (mean age: 23.9+/-1.7) and 50 females (mean age: 23.3+/-1.6) were included in a randomized, split-mouth, localized 21-day experimental gingivitis trial. Plaque index (PI), gingival index (GI), gingival crevicular fluid volume (GCF), and angulated bleeding score (AngBS) were recorded. Two subgroups were defined from the total study population (HR, LR) characterized by substantially different severity of gingival inflammation despite similar plaque accumulation rate. The study population was typed for interleukin-1 alpha (IL 1A+4845), interleukin-1 beta (IL-1B+3953, IL-1B-511), and IL-1 receptor antagonist (IL-1RN, intron 2 variable number tandem repeats) gene polymorphisms. Gene variants were analyzed by amplifying the polymorphic region using polymerase chain reaction, followed by restriction-enzyme digestion and agarose gel electrophoresis. RESULTS: Neither IL-1A+4845, IL-1B+3953, or the combined (IL 1A+4845 x 2 - IL-1B+3953 x 2) genotype was associated with clinical parameters in the overall population. IL-1RN was significantly associated with test quadrant PI (P= 0.046), GCF (P= 0.05), and GI (P= 0.018). The genotype distribution in HR and LR subjects was significantly different for IL-1RN (P= 0.045) and for IL-1B-511 (P= 0.023). CONCLUSION: The results of the present study suggest an association between IL-1RN polymorphism and subject-based clinical behavior of the gingiva in response to de novo plaque accumulation, as well as a possible association between IL-1B-511 polymorphism and gingivitis susceptibility. PMID- 15830638 TI - Comparative evaluation of decalcified and non-decalcified freeze-dried bone allografts in rhesus monkeys. I. Histologic findings. AB - BACKGROUND: Controversy exists regarding the relative merits of decalcified (DFDBA) and non-decalcified (FDBA) freeze-dried bone allografts when used in periodontal or other oral surgical procedures. METHODS: Under typical sedation, six rhesus monkeys had nylon mesh cylinders containing either DFDBA or FDBA implanted into surgically created vertical grooves on the facial aspects of all posterior quadrants. Each quadrant received three cylinders containing one type of bone, plus one empty cylinder (E) as negative control, and the full thickness flaps were closed to completely cover the cylinders. Cylinders were retrieved at 1, 2, and 3 months and processed for histologic evaluation. Photomicrographs taken at 8x were randomly evaluated using a histometric grid point counting technique for new bone and old bone within the chambers, and the data were analyzed with analysis of variance plus post-tests. RESULTS: The nylon chambers and their contents were well-tolerated by the tissues. FDBA chambers contained more new bone and total bone than either the DFDBA or E chambers at all time periods (P <0.05). DFDBA was not statistically significantly different than E at any time period. FDBA had less old bone than DFDBA at 3 months (P<0.05). FDBA and DFDBA had more total bone (grafted plus new) present than E at all time periods (P<0.05). CONCLUSION: These results suggest that FDBA may stimulate earlier, more rapid, and more substantial new bone formation than DFDBA in a monkey jaw defect model system. PMID- 15830639 TI - Non-surgical periodontal therapy with adjunctive topical doxycycline: a double masked, randomized, controlled multicenter study. II. Microbiological results. AB - BACKGROUND: Topical application of active substances offers an additional option in periodontal therapy. The aim of this study was to examine the influence of the administration of a novel, biodegradable 14% doxycycline gel on microbiological findings, in connection with scaling and root planing. METHODS: One hundred ten patients in three centers (Frankfurt and Heidelberg, Germany; Nijmegen, The Netherlands) with moderate to advanced periodontitis were evaluated in this randomized, double-masked, split-mouth clinical trial. In each patient, three test teeth were randomly assigned to one of three treatment modalities: 1) scaling and root planing (SRP) alone; 2) SRP with subgingival placebo gel (VEH); or 3) SRP and 14% doxycycline gel (DOX). Subgingival plaque samples were analyzed at baseline and 3 and 6 months after therapy for Actinobacillus actinomycetemcomitans (A.a.), Tannerella forsythensis (T.f.), Porphyromonas gingivalis (P.g.), and Treponema denticola (T.d.) using a RNA probe. Samples from 10 patients were tested for resistance against doxycycline, amoxicillin/clavulanic acid, cefoxitin, clindamycin, and metronidazole using agar diffusion testing. RESULTS: The largest decrease in pathogens was found after 3 months, with the most pronounced differences between DOX and SRP (P <0.05). At 6 months, pathogens were still reduced markedly in all groups. Treatment results were consolidated for VEH and DOX, with a slight deterioration for SRP (DOX versus SRP: P <0.001). Resistance was observed to amoxycillin/clavulanic acid, cefoxitin, clindamycin, and metronidazole (four isolates) but not to doxycycline. CONCLUSION: The addition of subgingival instillation of a 14% doxycycline gel resulted in pronounced reduction of periodontal pathogens after 3 months and stabilizing results up to 6 months after therapy. Resistance to doxycycline was not induced. PMID- 15830640 TI - Increased interleukin-18 in patients with juvenile idiopathic arthritis and early attachment loss. AB - BACKGROUND: Patients with juvenile idiopathic arthritis (JIA) have been shown to have incipient attachment loss (AL) more frequently than systemically healthy individuals. This study investigated neutrophil activity and proinflammatory cytokines in these patients and aged-matched controls. METHODS: Elastase activity, measured with a low molecular weight substrate (S-2484), and interleukin-1beta (IL-1beta), measured with the enzyme-linked immunosorbent assay (ELISA), were analyzed in the gingival fluid of 38 patients with JIA and 29 controls. IL-1beta and interleukin-18 (IL-18) were measured with ELISA in the serum of the same groups. Subgingival plaque was analyzed by DNA probes to detect 12 bacteria. RESULTS: Significantly elevated serum levels of IL-1beta and IL-18 were found in the JIA group. No differences were found in the gingival fluid levels of elastase and IL-1beta between groups, or in the frequency of subjects positive to most of the bacteria analyzed, except F. nucleatum, C. rectus, P. micros, and S. intermedius, which were significantly more frequent in the control group. When the JIA group was subdivided according to the presence/absence of AL, IL-18 was significantly increased in the JIA subgroup with AL compared to those without it. There were no differences in the subgingival microbiota between the subgroups. CONCLUSION: The findings of increased serum IL-18 and IL-1beta in patients with JIA accompanied by a similar subgingival microbiota suggest that the increased frequency of incipient attachment loss observed in these patients might be due to their altered systemic inflammatory response, making them more susceptible to periodontal disease. PMID- 15830641 TI - Collagen fiber orientation in human peri-implant bone around immediately loaded and unloaded titanium dental implants. AB - BACKGROUND: The main factor in determining the mechanical properties of bone is the collagen configuration. METHODS: This study investigated the birefringence in human bone around loaded and unloaded titanium dental implants to evaluate the collagen fiber orientation using circularly polarized light (CPL) and scanning electron microscopy (SEM). A total of 10 titanium dental implants, five immediately loaded and five unloaded, were used. The birefringence measurements were performed on digitized images of both loaded and unloaded implants. All images detected at 50x were measured using a software image analysis. RESULTS: In the bone around loaded implants, the transverse collagen fiber area was 45,481+/ 3,037 pixel2 (mean+/-SD), while the area of longitudinal collagen fibers was 13,676+/-2,232 pixel2 (mean+/-SD). In the unloaded implants, the transverse collagen fiber area was 32,174+/-2,554 pixel2 (mean+/-SD), while the area of longitudinal collagen fibers was 89,073+/-1,960 pixel2 (mean+/-SD). The CPL measurements of the birefringence for transverse collagen fibers of loaded versus unloaded implants indicated that the differences were statistically significant (P <0.05). The results for the longitudinal collagen fibers of loaded versus unloaded implants were also statistically significant (P <0.05). CONCLUSIONS: In the bone around loaded dental implants, transverse collagen fibers were more abundant, while in the unloaded implants, collagen fibers run more longitudinally. The load seemed to determine the collagen fiber orientation. PMID- 15830642 TI - Coping with stress: its influence on periodontal therapy. AB - BACKGROUND: Individual stress coping strategies appear to influence periodontal disease: individuals with inadequate stress behavior may be at greater risk of developing disease. The purpose of this 24-month prospective study was to examine the influence of different coping behaviors on a non-surgical periodontal therapy and on the course of periodontal disease. METHODS: In 80 patients with chronic periodontitis, a non-surgical periodontal treatment was conducted after their individual stress coping strategies had been recorded. After 2 years of regular maintenance, their periodontal condition was evaluated. The stress coping questionnaire was used to obtain psychodiagnostic data. Clinical attachment loss (CAL) served as the clinical parameter. RESULTS: Patients with a defensive coping style had statistically significant poorer attachment values (P= 0.000) after 2 years compared to patients with other coping behaviors. The percentage of sites with slight to moderate CAL (<5 mm) was significantly less in patients with a defensive coping style than in patients with other coping strategies (P = 0.000). The number of sites with severe advanced CAL (>5 mm) was significantly correlated with a suppressive coping style (P= 0.0001). None of the individual stress coping styles revealed significant overall changes over time. The subtest of drug use (alcohol, nicotine, tranquilizers) as well as changes in this subtest over time were significantly correlated with the CAL (P = 0.003); an increase in the t value of the subtest of drug use was accompanied by a significant increase in CAL. CONCLUSIONS: The results of this study show that passive coping strategies were more pronounced in advanced disease as well as in cases of poor response to a non-surgical periodontal treatment, whereas patients with active coping modes had milder disease and a more favorable course of treatment. Thus, maladaptive behavior, especially in association with behavior-related risk factors such as smoking, are of great importance in the medical history, treatment, and maintenance of patients with periodontal disease. PMID- 15830643 TI - Individual and combined effects of selective cyclooxygenase-2 inhibitor and omega 3 fatty acid on endotoxin-induced periodontitis in rats. AB - BACKGROUND: The present study was planned to evaluate the individual and combined effects of selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib, and omega-3 fatty acid on the gingival tissue levels of prostaglandin E2 (PGE2), prostaglandin F2alpha (PGF2alpha), leukotriene B4 (LTB4), and platelet activating factor (PAF) in endotoxin-induced periodontitis in rats. METHODS: Experimental periodontitis was induced by repeated injection of Escherichia coli endotoxin (LPS). Forty-four adult male Sprague-Dawley rats were divided into five study groups: saline control, LPS, celecoxib, omega-3 fatty acid, and combination celecoxib and omega-3 fatty acid. Celecoxib and omega-3 fatty acid were given either as a single agent or as a combination therapy during 14 days of the study period. At the end of the 2-week protocol, the rats were sacrificed, the gingival tissues were dissected and extracted, and the extracts were analyzed for PGE2, PGF2alpha, and LTB4 levels by enzyme immunoassay and for PAF levels by radioimmunoassay. The defleshed jaws were analyzed morphometrically for alveolar bone loss. Data were evaluated statistically by using parametric tests. RESULTS: LPS injection resulted in significantly more bone loss than the saline controls (P<0.05) and significant elevations in the gingival tissue levels of all the analyzed mediators except PGF2alpha. Individual administration of celecoxib revealed significant reductions in PGE2 and PAF levels (P <0.05), while omega-3 fatty acid provided significant reduction in PGE2, PGF2alpha, and LTB4 levels compared to the LPS group (P <0.05). Combined administration of celecoxib and omega-3 fatty acid exhibited significantly lower values than those of the LPS group in all the analyzed membrane phospholipid mediators (P <0.05), which approximated the levels in the saline control group (P>0.05). CONCLUSIONS: The results of the present study indicate that celecoxib and omega-3 fatty acid, when used individually, show a rather partial effect on the control of the analyzed mediators, but when combined they show a synergic effect and provide significant reductions in the gingival tissue levels of PGE2, PGF2alpha, LTB4, and PAF in LPS induced experimental periodontitis. These findings may pioneer further clinical human studies investigating the possible place of celecoxib and omega-3 fatty acid in periodontal treatment. PMID- 15830644 TI - Alendronate therapy may be effective in the prevention of bone loss around titanium implants inserted in estrogen-deficient rats. AB - BACKGROUND: This study evaluated whether alendronate (ALD) influences bone healing around titanium implants inserted in ovariectomized rats and whether it provides a residual effect after its withdrawal. METHODS: Bilateral ovariectomies were performed in 87 Wistar rats and one screw-shaped titanium implant was placed in the tibiae. The animals were divided into the following groups: group SHAM (N = 15): sham surgeries; group OVX (N = 15): ovariectomy; group AT (N = 15): OVX plus alendronate administration for 80 days; group AW (N = 14): OVX plus alendronate administration for 40 days; group ET (N = 14): OVX plus 17beta estradiol administration for 80 days; or group EW (N = 14): OVX plus 17beta estradiol administration for 40 days. Bone-to-implant contact (BIC), bone area (BA) within the limits of implant threads, and bone density in a 500 microm-wide zone lateral to the implant (BD) were obtained and measured for the cortical (zone A) and cancellous (zone B) regions. RESULTS: In zone A, data analysis showed no significant differences among the groups regarding BIC and BD (P >0.05), and a slight beneficial effect of estradiol on BA when compared with the OVX, EW, and AW groups (P <0.05). In zone B, OVX negatively impacted bone healing around the implants, resulting in reduced BA and BD (P<0.05). ALD (continuous/interrupted) and estradiol (only continuous) positively affected BIC, BA, and BD, resulting in values at the same level as the control group (SHAM). CONCLUSIONS: ALD may prevent the negative influence of estrogen deficiency on bone healing around titanium implants inserted in OVX rats. This positive effect, in contrast to estradiol, is sustained following its withdrawal. PMID- 15830645 TI - DNA probe identification of bacteria colonizing internal surfaces of the implant abutment interface: a preliminary study. AB - BACKGROUND: Currently, there is limited knowledge concerning the specific genus and species of bacteria that may colonize internal surfaces of the implant abutment interface (IAI) of two-stage dental implants. The purpose of this study was to use DNA probe analysis to identify those periodontopathic bacteria that may inhabit the internal surfaces and healing abutment screw-threads of the IAI of dental implants in situ. METHODS: Following osseointegration, bacterial samples for DNA probe analysis were obtained from 54 two-stage hydroxyapatite plasma spray-coated implants in 32 patients. Using sterile paper points, samples were obtained from the IAI of 43 implants and the screw-threads of healing abutments in the other 11 implants. DNA probes were available to detect the following microbes: Actinobacillus actinomycetemcomitans, Tannerella forsythensis, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, and Treponema denticola. RESULTS: All samples taken from healing abutment screw-threads were negative for all target microbes. The aggregate percentage of positive results for each target microbe for samples from internal surfaces of the IAI were: 41.9%, A. actinomycetemcomitans, 60.5%, T. forsythensis; 44.2%, C rectus; 60.5%, E. corrodens; 48.8%, F. nucleatum; 46.5%, P. gingivalis; 55.8%, P. intermedia; and 51.2%, T. denticola. In addition, no significant differences were noted between colonization of individual microbial species when comparing anterior to posterior and maxillary to mandibular implant sites. CONCLUSIONS: Moderate to high levels of eight different periodontopathic microbes inhabiting the internal surfaces of the IAI of 43 two-stage implants in partially edentulous patients were identified by DNA probe analysis. The microbes colonized these surfaces within 25 days following the second stage surgery and placement of the healing abutment. In contrast, all samples obtained from screw-threads of 11 healing abutments were DNA probe negative. These findings appear to support those of other investigations demonstrating the translocation of bacteria from residual dentition to implants. PMID- 15830646 TI - A pilot study into measurements of markers of atherosclerosis in periodontitis. AB - BACKGROUND: Periodontitis may be a possible risk factor for atherosclerosis. The current pilot study explored arterial wall thickness and other variables associated with atherosclerosis in healthy subjects with and without periodontitis. METHODS: Patients with moderate (N = 34) and severe periodontitis (N = 15) and controls (N = 14) were recruited. Intima media thickness (IMT) of the common carotid arteries (CCA), internal carotid arteries (ICA), and bifurcations of carotid arteries (BCA) was estimated bilaterally using B-mode ultrasound. An overall IMT was calculated as the mean of these six measurements. C reactive protein (CRP), fibrinogen, and von Willebrand factor (vWf) were measured in plasma as indicators of systemic inflammation and atherosclerotic disease. Microalbuminuria was determined as a marker of endothelial cell dysfunction. RESULTS: IMT for CCA were 0.64, 0.68, and 0.69 mm for control, moderate, and severe periodontitis, respectively (not significant). IMT for BCA did not vary among groups. IMT of ICA was largest for severe periodontitis (0.81 mm); corresponding values for controls and moderate periodontitis were 0.58 and 0.55 mm, respectively (P= 0.023). Severe periodontitis patients had an overall IMT of 0.76 mm, while moderate periodontitis patients and controls had lower values (0.64 and 0.65 mm, respectively; P= 0.153). After adjusting for potential confounding factors, the increased IMT for ICA in severe periodontitis was also significant (Padj = 0.040). CRP (P= 0.020, Padj = 0.050) and vWf (P= 0.019, Padj = 0.013) were higher in periodontitis than controls; microalbuminuria was not different between groups. Power calculations suggest that a 4-fold expansion of the severe patient and control groups will result in a high chance (power level 80%) that a clinically significant association between the overall IMT and periodontitis will be observed. CONCLUSION: The present pilot study indicates that a full study investigating the relationship between periodontitis and atherosclerosis is warranted. PMID- 15830647 TI - Clinical and radiographic effects of enamel matrix derivative in the treatment of intrabony periodontal defects: a 12-month longitudinal placebo-controlled clinical trial in adult periodontitis patients. AB - BACKGROUND: This randomized, double-masked, placebo-controlled clinical trial evaluated the effect of enamel matrix derivative (EMD) on clinical and radiographic parameters of periodontal intrabony defects. METHODS: A split-mouth design was used in 16 chronic periodontitis patients who had similar defects (> or =6 mm of probing depth). Both groups underwent scaling and root planing and were acid-etched with EDTA. The test sites received the EMD solution and the controls a placebo. Clinical examinations of all 16 patients and radiographs of 14 patients were available at baseline and 6 and 12 months after surgery. Clinical outcomes included probing depth (PD) and clinical attachment level (CAL); radiographic analysis was performed using computerized linear measurements. Intergroup comparisons were performed by paired samples t test, and over time comparisons were made by general linear model (alpha = 0.05). RESULTS: A statistically significant improvement over time for PD and CAL and a decrease of the vertical component of the defect was detected in both groups. Comparisons between groups revealed at baseline a mean+/-SD value of CAL of 12.93+/-2.00 and 13.47+/-2.93 for test and control groups, respectively. These values decreased to 10.92+/-1.92 and 11.31+/-1.86 after 12 months for test and control. No statistically significant differences could be observed between groups. PD displayed similar results from 7.57+/-1.02 and 7.38+/-1.16 for test and control groups at baseline to 3.40+/-1.82 and 2.99+/-1.07 after 12 months. If the data are divided into smokers and non-smokers, no differences are observed. CONCLUSION: Use of EMD did not result in more improvement in clinical and radiographic parameters compared to the placebo. PMID- 15830648 TI - Severe pregnancy complication (preeclampsia) is associated with greater periodontal destruction. AB - BACKGROUND: Preeclampsia is a pregnancy-specific disease which is one of the leading causes of maternal and fetal morbidity and mortality. The etiology of preeclampsia remains elusive. The underlying pathology may be related to a generalized intravascular hyperinflammatory state. Since the placental histopathologic lesions of acute atherosis, seen in preeclampsia, bear resemblance to those seen in atherosclerosis, and recent evidence links atherosclerosis with chronic infection, we decided to investigate a possible link between preeclampsia and chronic periodontal infection. METHODS: Thirty primigravidas, 15 suffering from preeclampsia, had full mouth periodontal examinations. Gingival crevicular fluid samples were taken for laboratory immunological assessment. Similar examination and sampling were performed in 15 age- and maternal status-matched controls. RESULTS: Significantly higher periodontal probing depth and clinical attachment level scores were found in the preeclamptic group compared with controls (2.98 versus 2.11 and 3.33 versus 2.30, respectively). Plaque index, gingival index, and mean gingival overgrowth scores were very similar in both groups. Furthermore, prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-alpha, and interleukin (IL)-1beta levels were all significantly higher in the preeclamptic group (247.4 ngr/sample versus 89.1 ngr/sample, 8.03 pgr/sample versus 4.03 pgr/sample, and 175.7 pgr/sample versus 62.2 pgr/sample, respectively). CONCLUSION: The above results suggest a possible association between periodontal inflammation and preeclampsia. PMID- 15830649 TI - Coronally repositioning the cemento-enamel junction to address gingival margin discrepancies. AB - BACKGROUND: Gingival margin discrepancies, especially in the esthetic zone, can pose a challenge for the restorative dentist. Surgical clinical crown lengthening and orthodontic tooth movement are most often used to treat these situations. In this report, an additional surgical technique will be discussed with its indications for use. METHODS: Three cases are presented, each with a different type of gingival margin discrepancy. The surgical technique used was the same for all cases. It involved establishing a new, more coronally positioned cemento enamel junction (CEJ) through the use of a finishing bur and curets. The newly exposed root surface was then covered with connective tissue, thus effecting a movement of the gingival margin in a coronal position. RESULTS: Clinical outcomes resulted in a more natural soft tissue relationship between the treated tooth and the adjacent dentition. The three cases presented did not appear to have any negative periodontal or restorative consequences following the procedure. CONCLUSIONS: This report presents another technique which can be considered when the clinician is faced with the need to move the gingival margin in a coronal direction. Within the guidelines presented, this technique appears to be predictable, less complicated, and less time consuming to carry out when compared with other available options. PMID- 15830650 TI - Langerhans-cell histiocytosis: a clinical case without bone involvement. AB - BACKGROUND: Langerhans-cell histiocytosis (LCH) is a group of rare disorders histologically characterized by the proliferation of Langerhans cells. Multiple organs and systems may be involved by the disease. Typically, there is bone involvement and, less frequently, lesions may be found in the lungs, liver, lymph nodes, skin, and mucosae. Oral soft tissue lesions without bone involvement are rare. METHODS: We report a case of a 23-year-old man with LCH detected by oral soft tissue, cutaneous, and lung lesions. RESULTS: Due to the oral lesion diagnosis, important procedures were performed, which determined the staging of the disease. CONCLUSION: Gingival lesions could be one of the first manifestations of LCH. The periodontist should recognize and detect this important pathology with oral involvement. PMID- 15830651 TI - Alveolar bone necrosis and tooth exfoliation following herpes zoster infection: a review of the literature and case report. AB - BACKGROUND: Herpes zoster (HZ) presents as a cutaneous vesicular eruption in the area innervated by the affected sensory nerve, usually associated with severe pain. Oral manifestations of HZ appear when the mandibular or maxillary divisions of the trigeminal nerve are affected. METHODS: This is a case report of a 63-year old woman with HZ infection with trigeminal nerve involvement that led to a rapid loss of alveolar bone and exfoliation of two teeth. RESULTS: The initial intraoral examination showed redness of the alveolar mucosa and gingiva of the lower right quadrant with multiple well-delimited and painful erosive lesions affecting the attached gingiva around the teeth. Two weeks later, teeth number 27 (lower right canine) and 28 (lower right first premolar) had class III mobility, flow of purulent exudate from the gingival sulcus, and deep pockets (>11 mm). The radiological examination showed advanced alveolar bone loss around both teeth. The prognosis for teeth number 27 and 28 was considered hopeless, and they were extracted. Due to extensive necrosis there was no interdental alveolar bone. The case is presented with a review of clinical data from patients with trigeminal HZ infection associated with osteonecrosis or exfoliation of teeth previously reported in the literature. The mechanisms by which the HZ infection leads to the alveolar bone necrosis are discussed. CONCLUSIONS: Extensive osteonecrosis and exfoliation of teeth in the area innervated by the nerve affected by HZ has been reported after HZ infection. Clinicians should be aware of this possible outcome after a trigeminal HZ infection. PMID- 15830652 TI - Early detection and successful management of oral pemphigus vulgaris: a case report. AB - BACKGROUND: Pemphigus vulgaris (PV) is an autoimmune disease characterized by acantholysis in the epithelium. PV is often difficult to diagnose in the early stages, since the oral lesions are relatively nonspecific. We report on a case with a favorable outcome due to early diagnosis and effective treatment of oral lesions. METHODS: A 53-year-old Japanese woman presented with a month-long history of painful gingiva. PV was diagnosed using clinical, histopathological, and direct immunofluorescence criteria 1 month after the first visit. Evaluation of the circulating autoantibody titers to desmoglein (Dsg)1 and Dsg3 was conducted by enzyme-linked immunosorbent assay (ELISA). Occlusive steroid therapy (OST) using a topical corticosteroid (0.1% triamcinolone acetonide) was provided for 8 weeks. After OST was completed, a buccal frenectomy was performed to eliminate localized toothbrushing trauma. RESULTS: There were no adverse effects on wound healing after the procedure. No oral PV lesions were detectable at the 9 month reevaluation, and the patient remains in complete remission. The Dsg1 ELISA was negative while the Dsg3 ELISA was indeterminate at the first visit. The Dsg1 ELISA was also negative at reevaluation, but Dsg3 ELISA reactivity had increased 9 months after completion of treatment. CONCLUSION: Large scale clinical trials will be required to validate the clinical benefits of the OST treatment regimen, and further research is necessary to determine the importance of ELISA antibody examination in long-term management of oral PV. PMID- 15830653 TI - Prevalence and numbers of Escherichia coli O157 on bovine hides at a beef slaughter plant. AB - In this study, we investigated the prevalence and numbers of Escherichia coli O157 on bovine hides. Samples (n = 1,500) were collected over a 17-month period (30 samples per week) by sponge swabbing approximately 122-cm2 areas of the bovine rump of slaughtered cattle at an early stage of carcass processing (first legging). Sponge samples (n = 1,500) were stomached in buffered peptone water supplemented with novobiocin, directly plated on sorbitol MacConkey with Cefixime tellurite (SMAC-CT), enriched for 24 h, extracted by immunomagnetic separation, and plated onto SMAC-CT agar. Presumptive E. coli O157 colonies from SMAC-CT plates were confirmed by PCR for the presence of eaeA, hlyA, fliCh7, vt1, vt2, and portions of the rfb (O-antigen encoding) region of E. coli O157. Overall, E. coli O157 was recovered from 109 samples (7.3%) at concentrations ranging from less than 0.13 to 4.24 log CFU/100 cm2. PCR analysis revealed a wide diversity of genetic profiles among recovered isolates of verocytotoxigenic E. coli. Of the isolates recovered, 99 of 109 contained the attaching and effacing gene (eaeA) and the hemolysin gene (hlyA), and 78 of 109 had the flagellar H7 antigen encoding gene (fliCh7). Only 6 of 109 isolates contained both verotoxin-producing genes (vt1 and vt2); 91 of 109 contained the vt2 gene only, whereas 1 of 109 contained the vt1 gene only. The remaining 11 of 109 contained neither vt1 nor vt2. PMID- 15830654 TI - Effects of low-dose, low-penetration electron beam irradiation of chilled beef carcass surface cuts on Escherichia coli O157:H7 and meat quality. AB - Low-dose, low-penetration electron beam (E-beam) irradiation was evaluated for potential use as an antimicrobial intervention on beef carcasses during processing. The objectives of this study were (i) to assess the efficacy of E beam irradiation to reduce concentrations of Escherichia coli O157:H7 on a large beef surface and (ii) to evaluate the effect of the treatment on the sensory properties of the product. A 1-kGy dose of E-beam radiation reduced E. coli O157:H7 inoculated onto sections of cutaneous trunci at least 4 log CFU/cm2. In assessing organoleptic impact, flank steak was used as the model muscle. Flank steaks with various levels of penetration by radiation (5, 10, 25, 50, and 75%) were evaluated. None of the flank steak sensory attributes were affected (P > 0.05) by any penetration treatment. Ground beef formulations consisting of 100, 50, 25, 10, 5, and 0% surface-irradiated beef were tested. A trained sensory panel did not detect any difference between the control (0%) and either the 5 or 10% treatments. These results suggest that if chilled carcasses were subjected to low-dose E-beam irradiation, aroma and flavor of ground beef would not be impacted. The data presented here indicate that low-dose, low-penetration E-beam irradiation has potential use as an antimicrobial intervention on beef carcasses during processing and minimally impacts the organoleptic qualities of the treated beef products. PMID- 15830655 TI - Influence of acetic, citric, and lactic acids on Escherichia coli O157:H7 membrane lipid composition, verotoxin secretion, and acid resistance in simulated gastric fluid. AB - The effect of organic acid (acetic, citric, and lactic acids) adaptation at equivalent initial pH values (6.4 and 5.4) on changes in membrane lipid composition, verotoxin concentration, and acid resistance in simulated gastric fluid (pH 1.5, 37 degrees C) was determined for Escherichia coli O157:H7 ATCC 43895 (HEC) and an rpoS mutant of E. coli O157:H7 ATCC 43895 (RM, FRIK 816-3). For HEC, lactic acid-adapted (pH 5.4) cells had the greatest D-value (32.2 min) and acetic acid-adapted (pH 5.4) cells had the smallest D-value (16.6 min) in simulated gastric fluid. For RM, D-values of citric and acetic acid-adapted cells were similar to those for nonadapted cells grown at pH 7.3, but D-values increased from 13.1 to 27.9 min in lactic acid-adapted cells (from pH 7.3 to pH 5.4). For both strains, the ratio of cis-vaccenic to palmitic acids decreased for citric and lactic acid-adapted cells, but the ratio increased for acetic acid adapted cells at pH 5.4. Organic acid-adapted cells produced less total verotoxin than did nonadapted cells at approximately 10(8) CFU/ml. Extracellular verotoxin concentration proportionally decreased with decreasing pH for both HEC and RM. Changes in membrane lipid composition, verotoxin concentration, and acid resistance in HEC and RM were dependent on both pH and organic acid. Deletion of the rpoS gene did not affect these changes but did decrease acid resistance in citric acid-adapted cells. Results indicate that decreased membrane fluidity may have caused increased acid resistance and decreased verotoxin secretion. PMID- 15830656 TI - Development of green fluorescent protein-expressing bacterial strains and evaluation for potential use as positive controls in sample analyses. AB - Strains of enterohemorrhagic Escherichia coli O157:H7 and Salmonella Typhimurium were engineered to express the gene for a modified green fluorescent protein (GFP) and were evaluated for potential use as positive controls in sample analyses. The strains fluoresced when observed as colonies with a handheld UV lamp or as individual cells under a fluorescent microscope. The strains maintained their fluorescence following growth in three series of transfer experiments including 8 to 11 passages from broth to broth and twice for 15 consecutive transfers from broth onto Trypticase soy agar plates. Cultures also maintained stability in the ability to fluoresce when agar plates were refrigerated (4 degrees C) for up to 12 days. Growth characteristics of the GFP positive strains were comparable to those of corresponding control strains. The GFP-positive strains were successfully identified using rapid diagnostic methods and were differentiated from their corresponding non-GFP strains by pulsed-field gel electrophoresis but not by repetitive extragenic palindromic PCR. The GFP positive and the control strains were recovered successfully from individually inoculated food samples (Feta cheese, raw shrimp, cooked shrimp, and cooked crawfish). However, in one Feta cheese sample and one raw shrimp sample inoculated with combined GFP-positive and GFP-negative cultures, colonies of the GFP-positive strains were not observed under UV light; fluorescing cells in one of the inoculated samples (raw shrimp) were revealed by microscopy. In general, the isolates from the inoculated foods were GFP positive by microscopic examination; the pure isolates could also be restreaked onto Trypticase soy agar, and colonies could be visually examined under UV light. Because GFP strains are not known to occur naturally in the environment, the use of the Salmonella GFP positive strain may offer advantages as a positive control even when distinct and rare serotypes are available. The GFP-positive E. coli O157:H7 strain may also prove beneficial for use as a positive control strain for sample analyses. PMID- 15830657 TI - Survival of salmonella transformed to express green fluorescent protein on Italian parsley as affected by processing and storage. AB - To study the effect of processing and storage parameters on the survival of Salmonella on fresh Italian parsley, parsley bunches were dipped for 3 or 15 min in suspensions that were preequilibrated to 5, 25, or 35 degrees C and inoculated with Salmonella transformed to express enhanced green fluorescent protein. Loosely attached and/or associated, strongly attached and/or associated, and internalized and/or entrapped Salmonella cells were enumerated over 0, 1, and 7 days of storage at 25 degrees C and over 0, 1, 7, 14, and 30 days of storage at 4 degrees C using surface-plating procedures. Leaf sections obtained from samples after 0, 1, and 7 days of storage were examined using confocal scanning laser microscopy. Temperature of the dip suspension had little effect on the attachment and survival of Salmonella cells on parsley. Regardless of the temperature or duration of dip, Salmonella was internalized. Immersion for longer times resulted in higher numbers of attached and internalized cells. Microscopic observations supported these results and revealed Salmonella cells near the stomata and within cracks in the cuticle. Storage temperature had the greatest impact on the survival of Salmonella cells on parsley. When stored at 25 degrees C, parsley had a shelf life of 7 days, and Salmonella populations significantly increased over the 7 days of storage. For parsley stored at 4 degrees C, numbers of Salmonella cells decreased over days 0, 1, and 7. After 7 days of storage, there were no viable internalized Salmonella cells detected. Storage temperature represents an important control point for the safety of fresh parsley. PMID- 15830658 TI - Salmonella in dairy operations in the United States: prevalence and antimicrobial drug susceptibility. AB - Salmonella serotypes are important foodborne pathogens of humans that can be acquired through consumption of contaminated meat and dairy products. Salmonella infection also can be a significant animal health issue. As part of a national study of U.S. dairy operations conducted between March and September 2002, fecal samples were collected from representative cows in 97 dairy herds in 21 states and were cultured to determine the prevalence of Salmonella shedding. Salmonella was recovered from the feces of at least one cow in 30.9% of the herds. Overall, 7.3% of fecal samples were culture positive for Salmonella. The three most frequently recovered serotypes were Salmonella Meleagridis (24.1%), Salmonella Montevideo (11.9%), and Salmonella Typhimurium (9.9%). The susceptibilities of Salmonella isolates recovered were determined using a panel of 16 antimicrobial drugs. Salmonella isolates recovered from dairy cows had relatively little resistance to these antimicrobial agents; 83.0% of the isolates were susceptible to all antimicrobials tested. This study provides updated information on the prevalence and susceptibility patterns of Salmonella in dairy herds and on cow and herd characteristics. These data contribute to our understanding of the ecology of Salmonella in the dairy farm environment. PMID- 15830659 TI - Thermal resistance of Salmonella enterica serotypes, Listeria monocytogenes, and Staphylococcus aureus in high solids liquid egg mixes. AB - Decimal reduction times (D-values) were determined for Salmonella enterica serotypes, Listeria monocytogenes, and Staphylococcus aureus in two high solids egg mixes designated A and B (water activity [a(w)] = 0.76 and 0.82; solids = 53.12 and 52.63%; pH = 5.09 and 5.29; viscosity = 183 and 119 centipoise/s, respectively) using a low-volume (0.06 ml) sealed glass capillary tube procedure. For Salmonella, D-values ranged from 0.035 (70 degrees C) to 0.193 min (64 degrees C) in product A and from 0.048 to 0.193 min in product B. For Listeria, D values ranged from 0.133 (70 degrees C) to 0.440 min (64 degrees C) in product A and from 0.074 to 0.364 min in product B. For Staphylococcus, D-values ranged from 0.332 (70 degrees C) to 1.304 min (64 degrees C) in product A and from 0.428 to 1.768 min in product B. For Listeria, the D-values of all heating temperatures were significantly higher (P < 0.01) in product A than in product B. The similar trend was also observed for Salmonella and Staphylococcus but only at 66 degrees C for Salmonella and 64 degrees C for Staphylococcus. Greater temperature dependence was observed for Salmonella inactivation in the low a(w) and low pH product (A), while the product (B) with the higher a(w) and pH had greater temperature dependence for Listeria. Compared across both egg mixes and all heating temperatures, the Staphylococcus strains were from 6.2 to 11.7 times more heat resistant than S. enterica serotypes and from 2.2 to 7.5 times more heat resistant than L. monocytogenes. PMID- 15830660 TI - Inactivation of Salmonella enterica serovar Enteritidis on shell eggs by ozone and UV radiation. AB - The presence of Salmonella enterica serovar Enteritidis in shell eggs has serious public health implications. Several treatments have been developed to control Salmonella on eggs with mixed results. Currently, there is a need for time saving, economical, and effective egg sanitization treatments. In this study, shell eggs externally contaminated with Salmonella (8.0 x 10(5) to 4.0 x 10(6) CFU/g of eggshell) were treated with gaseous ozone (O3) at 0 to 15 lb/in2 gauge for 0 to 20 min. In other experiments, contaminated shell eggs were exposed to UV radiation at 100 to 2,500 microW/cm2 for 0 to 5 min. Treatment combination included exposing contaminated eggs to UV (1,500 to 2,500 microW/cm2) for 1 min, followed by ozone at 5 lb/in2 gauge for 1 min. Eggs that were (i) noncontaminated and untreated, (ii) contaminated and untreated, and (iii) contaminated and treated with air were used as controls. Results indicated that treating shell eggs with ozone or UV, separately or in combination, significantly (P < 0.05) reduced Salmonella on shell eggs. For example, contaminated eggs treated with ozone at 4 to 8 degrees C and 15 lb/in2 gauge for 10 min or with UV (1,500 to 2,500 microW/cm2) at 22 to 25 degrees C for 5 min produced 5.9- or 4.3-log microbial reductions or more, respectively, when compared with contaminated untreated controls. Combinations including UV followed by ozone treatment resulted in synergistic inactivation of Salmonella by 4.6 log units or more in about 2 min of total treatment time. Salmonella was effectively inactivated on shell eggs in a short time and at low temperature with the use of a combination of UV radiation and ozone. PMID- 15830661 TI - Growth of Salmonella enterica serovar Enteritidis in albumen and yolk contents of eggs inoculated with this organism onto the vitelline membrane. AB - By using an in vitro model simulating the potential opportunities for Salmonella enterica serovar Enteritidis (SE) to proliferate within eggs contaminated with this organism following oviposition, we investigated growth of SE in eggs. Seventy to 140 CFU of one of three SE strains originating either from egg contents, chicken meat, or a human infection were experimentally inoculated onto the vitelline membrane of eggs collected from specific-pathogen-free flocks of chickens and incubated at 25 degrees C. SE organisms were detected in 6 of 71 yolk contents of the eggs inoculated with any of the test strains attaining levels ranging from 2.0 x 10(2) to 4.2 x 10(8) CFU/ml by day 6. The organisms were also detected in the albumen from 38 of 55 eggs tested, growing to levels ranging from 1.0 x 10(2) to 4.3 x 10(8) CFU/ml by day 6 after inoculation. An additional three yolk contents and 15 albumen samples were culture positive for SE following enrichment. There was no correlation between the number of the organisms in the yolk contents and that in the albumen from each of the eggs. When 73 to 91 CFU of the egg strain were inoculated into samples of separated albumen obtained from eggs that were stored at 4 degrees C for 1 to 4 weeks or at 25 degrees C for 1 week, slight growth (3.0 x 10(2) to 7.4 x 10(3) CFU/ml) was found in only 3 of the 60 albumen samples by day 6 after inoculation, but the organisms were recovered from 52 samples following enrichment. The results suggest that the environment on or near the vitelline membrane can be conducive to SE proliferation over time. PMID- 15830662 TI - Prevalence and diversity of Campylobacter jejuni in pig herds on farms with and without cattle or poultry. AB - The prevalence and diversity of Campylobacter jejuni was investigated in pig herds on farms with and without cattle or poultry production. A bacteriological screening of pig cecal samples from 247 finisher herds was carried out at the slaughter-house. Subsequently, a follow-up study was conducted in 24 herds (either with or without prior C. jejuni isolation from pigs) in which fecal samples were collected from pigs and, if present, cattle and poultry. Samples were analyzed for presence of Campylobacter, and subsequent analysis included species identification, serotyping, and, for selected strains, pulsed-field gel electrophoresis typing. In the slaughterhouse screening, C. jejuni was isolated from pigs in 21 (8.5%) herds, but no significant difference in prevalence was found between herd types (pigs, pigs and cattle, pigs and poultry). At the slaughterhouse, C. jejuni and Campylobacter coli prevalence in pigs was 2.3 and 90.1%, respectively. In the follow-up study, herd prevalence of C. jejuni was 8.3%, whereas C. jejuni and C. coli were isolated from 0.8 and 92.0% of pigs, respectively. In mixed production herds, C. jejuni predominated in cattle (42.7%) and poultry (31.6%), whereas C. jejuni was only isolated from 1.3 to 2.5% of pigs in these herds. There were no significant differences in C. jejuni or C. coli prevalence in pigs, cattle, and poultry between herds with and without prior C. jejuni isolation at the slaughterhouse. Pulsed-field gel electrophoresis typing did not yield evidence of C. jejuni transmission between cattle or poultry and pigs in mixed production herds. In contrast, pulsed-field gel electrophoresis analysis showed indistinguishable serotypes of C. coli in pigs and cattle in two herds. Verification of C. jejuni-positive pig samples showed that individual pigs can excrete high levels of C. jejuni and that mixed infection with C. jejuni and C. coli was common in C. jejuni-positive pigs. The results of our study suggest that transmission of C. jejuni between pigs and cattle or poultry in mixed production herds occurs infrequently. Detection of indistinguishable C. coli isolates in two herds, however, might indicate the existence of low-level transmission between pigs and cattle in herds of mixed production. PMID- 15830663 TI - Development of a surface plasmon resonance-based immunoassay for Listeria monocytogenes. AB - A polyclonal antibody was produced against Internalin B (InlB)-enriched extract and used to develop an inhibition assay to detect Listeria monocytogenes cells in solution using surface plasmon resonance. The gene sequence encoding for the InlB protein was cloned into a Qiagen pQE-60 vector, expressed in Escherichia coli, and purified by immobilized metal affinity chromatography. Protein G-purified anti-InlB-enriched extract polyclonal antibody was incubated with various concentrations of L. monocytogenes cells and subsequently injected over a purified-recombinant InlB (rInlB)-immobilized CM5 sensor chip surface. A decrease in antibody binding response was observed with increasing L. monocytogenes cell concentrations. Intraday and interday assay variability studies were carried out to evaluate precision and reproducibility. The assay had a limit of detection of less than 2 x 10(5) cells per ml and could be successfully reproduced with coefficients of variation of between 2.5 and 7.7%. PMID- 15830664 TI - Predicting heat inactivation of Listeria monocytogenes under nonisothermal treatments. AB - The aim of this study was to find a model that accurately predicts the heat inactivation of Listeria monocytogenes (ATCC 15313) at constantly rising heating rates (0.5 to 9 degrees C/min) in media of different pH values (4.0 to 7.4). Survival curves of L. monocytogenes obtained under isothermal treatments at any temperature were nearly linear. Estimations of survival curves under nonisothermal treatments obtained from heat resistance parameters of isothermal treatments adequately fit experimental values obtained at pH 4.0. On the contrary, survivors were much higher than estimations at pH 5.5 and 7.4. The slower the heating rate and the longer the treatment time, the greater the differences between the experimental and estimated values. An equation based on the Weibullian-like distribution, log S(t) = (t/delta)p, accurately described survival curves of L. monocytogenes obtained under nonisothermal conditions within the range of heating rates investigated. A nonlinear relationship was observed between the scale parameter (delta) and the heating rate, which allowed the development of an equation capable of predicting the inactivation rate of L. monocytogenes under nonisothermal treatments at pH 5.5 and 7.4. The model predictions were a good fit to the measured data independent of the magnitude of the thermotolerance increase. This work might contribute to the increase in safety of those food products that require long heating lag phases during the pasteurization process. PMID- 15830665 TI - Inactivation of barotolerant Listeria monocytogenes in sausage by combination of high-pressure processing and food-grade additives. AB - Food-grade additives were used to enhance the efficacy of high-pressure processing (HPP) against barotolerant Listeria monocytogenes. Three strains of L. monocytogenes (Scott A, OSY-8578, and OSY-328) were compared for their sensitivity to HPP, nisin, tert-butylhydroquinone (TBHQ), and their combination. Inactivation of these strains was evaluated in 0.2 M sodium phosphate buffer (pH 7.0) and commercially sterile sausage. A cell suspension of L. monocytogenes in buffer (10(9) CFU/ml) was treated with TBHQ at 100 ppm, nisin at 100 IU/ml, HPP at 400 MPa for 5 min, and combinations of these treatments. Populations of strains Scott A, OSY-8578, and OSY-328 decreased 3.9, 2.7, and 1.3 log with HPP alone and 6.4, 5.2, and 1.9 log with the HPP-TBHQ combination, respectively. Commercially sterile sausage was inoculated with the three L. monocytogenes strains (10(6) to 10(7) CFU/g) and treated with selected combinations of TBHQ (100 to 300 ppm), nisin (100 and 200 ppm), and HPP (600 MPa, 28 degrees C, 5 min). Samples were enriched to detect the viability of the pathogen after the treatments. Most of the samples treated with nisin, TBHQ, or their combination were positive for L. monocytogenes. HPP alone resulted in a modest decrease in the number of positive samples. L. monocytogenes was not detected in any of the inoculated commercial sausage samples after treatment with HPP-TBHQ or HPP-TBHQ nisin combinations. These results suggest that addition of TBHQ or TBHQ plus nisin to sausage followed by in-package pressurization is a promising method for producing Listeria-free ready-to-eat products. PMID- 15830666 TI - An assessment of pasteurization treatment of water, media, and milk with respect to Bacillus spores. AB - This study evaluated the ability of spore-forming Bacillus spp. to resist milk pasteurization conditions from 72 to 150 degrees C. Spores from the avirulent surrogate Sterne strain of Bacillus anthracis, as well as a representative strain of a common milk contaminant that is also a pathogen, Bacillus cereus ATCC 9818, were heated at test temperatures for up to 90 min in dH2O, brain heart infusion broth, or skim milk. In skim milk, characteristic log reductions (log CFU per milliliter) for B. anthracis spores were 0.45 after 90 min at 72 degrees C, 0.39 after 90 min at 78 degrees C, 8.10 after 60 min at 100 degrees C, 7.74 after 2 min at 130 degrees C, and 7.43 after 0.5 min at 150 degrees C. Likewise, log reductions (log CFU per milliliter) for viable spores of B. cereus ATCC 9818 in skim milk were 0.39 after 90 min at 72 degrees C, 0.21 after 60 min at 78 degrees C, 7.62 after 60 min at 100 degrees C, 7.37 after 2 min at 130 degrees C, and 7.53 after 0.5 min at 150 degrees C. No significant differences (P < 0.05) in thermal resistance were observed for comparisons of spores heated in dH2O or brain heart infusion broth compared with results observed in skim milk for either strain tested. However, spores from both strains were highly resistant (P < 0.05) to the pasteurization temperatures tested. As such, pasteurization alone would not ensure complete inactivation of these spore-forming pathogens in dH2O, synthetic media, or skim milk. PMID- 15830667 TI - Monochloramine versus sodium hypochlorite as antimicrobial agents for reducing populations of bacteria on broiler chicken carcasses. AB - Studies were conducted to compare the effect of sodium hypochlorite (SH) versus monochloramine (MON) on bacterial populations associated with broiler chicken carcasses. In study 1, nominal populations (6.5 to 7.5 log CFU) of Escherichia coli, Listeria monocytogenes, Pseudomonas fluorescens, Salmonella serovars, Shewanella putrefaciens, and Staphylococcus aureus were exposed to sterilized chiller water (controls) or sterilized chiller water containing 50 ppm SH or MON. SH at 50 ppm eliminated all (6.5 to 7.5 log CFU) viable E. coli, L. monocytogenes, and Salmonella serovars; 1.2 log CFU of P. fluorescens; and 5.5 log CFU of S. putrefaciens. MON eliminated all (6.5 to 7.5 log CFU) viable E. coli, L. monocytogenes, S. putrefaciens, and Salmonella serovars and 4.2 log CFU of P. fluorescens. In study 2, chicken carcasses were inoculated with P. fluorescens or nalidixic acid-resistant Salmonella serovars or were temperature abused at 25 degrees C for 2 h to increase the populations of naturally occurring E. coli. The groups of Salmonella serovar-inoculated or temperature-abused E. coli carcasses were immersed separately in pilot-scale poultry chillers and exposed to tap water (controls) or tap water containing 20 ppm SH or 20 ppm MON for 1 h. The P. fluorescens-inoculated group was immersed in pilot-scale poultry chillers and exposed to tap water (controls) or tap water containing 50 ppm SH or 50 ppm MON for 1 h. Carcasses exposed to the SH treatment had nominal increases (0.22 log CFU) in E. coli counts compared with controls, whereas exposure to MON resulted in a 0.89-log reduction. Similarly, average nalidixic acid-resistant Salmonella serovar counts increased nominally by 34% (41 to 55 CFU/ml) compared with controls on carcasses exposed to SH, whereas exposure to MON resulted in an average nominal decrease of 80% (41 to 8 CFU/ml). P. fluorescens decreased by 0.64 log CFU on carcasses exposed to SH and decreased by 0.87 log CFU on carcasses exposed to MON. In study 3, SH or MON was applied to the chiller in a commercial poultry processing facility. E. coli counts (for carcass halves emerging from both saddle and front-half chillers) and Salmonella prevalence were evaluated. Data from carcasses exposed to SH during an 84-day historical (Hist) and a 9-day prepilot (Pre) period were evaluated. Other carcasses were exposed to MON and tested during a 27-day period (Test). E. coli counts for samples collected from the saddle chiller were 25.7, 25.2, and 8.6 CFU/ml for Hist, Pre, and Test, respectively. E. coli counts for samples collected from the front-half chiller were 6.7, 6.9, and 2.5 CFU/ml for Hist, Pre, and Test, respectively. Salmonella prevalence was reduced from 8.7% (Hist + Pre) to 4% (Test). These studies indicate that MON is superior to SH in reducing microbial populations in poultry chiller water. PMID- 15830668 TI - An evaluation of raw milk microorganisms as markers of on-farm hygiene practices related to milking. AB - Dairy farm hygiene audits were undertaken at 24 farms during summer and winter and the results compared with transformed bacterial indicator levels in raw milk samples collected during each audit. The bacterial indicators measured were total viable counts, Escherichia coli, coliforms, Bacillus spp., Bifidobacteria spp., and Pseudomonas spp. The results of initial comparisons using Pearson product moment correlation coefficients showed presumptive relationships between some bacterial groups and the subjective quantitative audit scores. When investigated further using linear regression, the presumptive relationships were found to be influenced by external factors. Possible reasons for the low correlations between on-farm hygiene and bacterial indicator counts in raw milk were further investigated. Measurements of the uncertainty associated with the bacteriological results were undertaken and revealed geometric relative standard deviations that ranged from 0.019 to 1.05. Toward the higher end of this scale, the uncertainty associated with the laboratory estimations of bacterial numbers may have been large enough to blur hygiene score-marker bacteria relationships. The samples obtained from on-farm raw milk storage tanks were representative of the whole tank contents and not a significant source of error. Although total bacterial counts are widely acknowledged by the milk industry as not always giving a true measure of on-farm hygiene during milking, we were unable to find any marker bacteria that showed consistently higher correlations and were thus better suited as indicators of on-farm hygiene. PMID- 15830669 TI - Microbiological quality of sous and tamarind, traditional drinks consumed in Jordan. AB - This study was conducted to evaluate the microbiological quality of sous (a drink prepared by extracting dried roots of Glycyrrhiza glabra) and tamarind (a drink prepared by infusing Tamarindus indica dried pulp), traditional drinks consumed in Jordan. Twenty-one samples of sous and 44 samples of tamarind were collected from the local market in Amman, Jordan. Water is the major component of the drinks. Sous drink is characterized by having an alkaline pH (range, 6.6 to 9.9; mean, 8.6), whereas tamarind drink has an acidic pH (range, 1.8 to 3.7; mean, 2.8). The drinks are not processed for safety before serving, and at some vendors drinks are not properly refrigerated. The mean counts for aerobic bacteria, lactic acid bacteria, and yeasts in sous drink samples were 5.9, 5.0, and 3.8 log CFU/ml, respectively; those in tamarind drink samples were 4.0, <1, and 5.8 log CFU/ml, respectively. The lactic acid bacteria isolated were Enterococcus raffinosus, Enterococcus hirae, Enterococcus durans, Lactobacillus acidophilus, and Lactobacillus buchneri. The yeast isolates in sous drink were from the genera Candida, Filobasidium, Hanseniaspora, Lodderomyces, Pichia, and Williopsis, and those in tamarind drink were from Arthroascus, Brettanomyces, Candida, Debaromyces, Filobasidiella, Hanseniaspora, Klavispora, Lodderomyces, Pichia, Saccharomycodes, Trichosporon, and Zygosaccharomyces. Enterobacteriaceae were detected in two sous samples and were identified as Enterobacter sakazakii and Erwinia sp., and in two tamarind samples and were identified as Citrobacter freundii and Klebsiella pneumoniae. Salmonella was detected in one sous and one tamarind sample. Pseudomonas aeruginosa was detected in only one sous sample. These findings highlight the importance of application of hygienic practices throughout preparation and vending of drinks, starting with raw ingredients and continuing through preparation, storage, display, and serving. PMID- 15830670 TI - Effect of ozone pretreatment on fish storage life at low temperatures. AB - The purpose of the present study was to determine the effect of ozone pretreatment of live fish on their shelf life and storage characteristics during storage at 0 and 5 degrees C. The evaluation included sensory, chemical, physical, and bacteriological tests at set intervals throughout a storage period of 30 days under two temperature regimes (0 and 5 degrees C). The results of the sensory tests showed that ozone pretreatment of live tilapia prolonged their shelf life by 12 days and improved their quality characteristics during storage at 0 degrees C for 30 days. These results were corroborated by the bacteriological tests. The sensory qualities of ozone-treated tilapia stored at 5 degrees C became unacceptable 3 days later than those of control fish. Although the most important factor affecting the fish shelf life is storage temperature, the combination of ozone pretreatment with storage at 0 degrees C appears to be a promising means of prolonging the shelf life. PMID- 15830671 TI - Consumption, knowledge, and handling of raw meat: a representative cross sectional survey in Germany, March 2001. AB - Outbreaks of gastroenteritis have been repeatedly associated with the consumption of raw meat and raw meat products. To assess the potential of this problem in Germany, frequency of consumption and knowledge about and handling of raw meat during food preparation in the German population was estimated. We conducted a population-based nationwide cross-sectional survey in private households in March 2001 by random telephone numbers. Consenting German-speaking persons were randomly chosen among household members for standardized interviews. Proportions were standardized by age, sex, and region (eastern and western Germany). A total of 510 people were interviewed (response 45.1%), including 62.3% women and 31.8% considered at high risk (children younger than 10 years old, elderly more than 65 years old, and chronically ill persons). The mean age was 45.7 years. Of those who ate raw ground meat at least once a week, 17.8% lived in eastern Germany and 5.1% in western Germany, (chi-square test, P < 0.001). Teewurst was eaten by 50.8% of all respondents and 53.8% of respondents in the high-risk group. That salami or Teewurst were raw meat products was known to 58.1 and 36.3% of the respondents, respectively. Respondents reported not washing their hands (46.6%) or cleaning their cutting boards with soap (48.1%) after preparing raw meat. A total of 10.3% of the respondents had diarrhea during the last 3 months. Although knowledge is poor, consumption of raw meat products is widespread in Germany, even among high-risk persons. More information is necessary on which products contain raw meat, the potential risk of diarrhea after raw meat products are consumed, and appropriate handling of raw meat products. PMID- 15830672 TI - Chemical composition and antimicrobial activity of Rosmarinus officinalis L. essential oil obtained via supercritical fluid extraction. AB - The chemical composition and antimicrobial activity of essential oil-rich fractions obtained by supercritical CO2 extraction from Rosmarinus officinalis L. were investigated. Gas chromatography-mass spectroscopy analysis of these fractions resulted in the identification of 33 compounds of the essential oil. The main components of these fractions were alpha-pinene, 1,8-cineole, camphor, verbenone, and borneol, constituting ca. 80% of the total oil. The antimicrobial activity was investigated by the disc diffusion and broth dilution methods against six microbial species, including gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa), a yeast (Candida albicans), and a fungus (Aspergillus niger). All of the essential oil-rich fractions obtained showed antimicrobial activity against all of the microorganisms tested, with inhibition zones and minimal bactericidal and fungicidal concentration values in the range of 17 to 33 mm and 2.25 to 0.25 mg/ml, respectively. The most active fraction was the one obtained in experiment 4 (4% ethanol as modifier; extraction pressure, 25 MPa; extraction temperature, 60 degrees C). S. aureus was found to be the most sensitive bacteria to the rosemary extracts, whereas the least susceptible was A. niger. alpha-Pinene, 1,8-cineole, camphor, verbenone, and borneol standards also showed antimicrobial activity against all the microorganisms tested, borneol being the most effective followed by camphor and verbenone. In that way, it was confirmed that essential oil from experiment 4, with the best antimicrobial activity, presented the highest quantity of camphor, borneol, and verbenone. PMID- 15830673 TI - Factors affecting the microbiological condition of the deep tissues of mechanically tenderized beef. AB - Whole or halved top butt prime beef cuts were treated in two types of mechanical tenderizing machines that both pierced the meat with thin blades but that used blades of different forms. Aerobes on meat surfaces and in the deep tissues of cuts after treatments were counted. When cuts were treated at a laboratory using a Lumar machine, the contamination of deep tissues increased significantly (P < 0.01) with increasing numbers of aerobic bacteria on meat surfaces and decreased significantly (P < 0.001) with increasing distance from the incised surface. However, contamination did not increase significantly (P > 0.1) with repeated incising of the meat. When halved cuts were incised one or eight times using a commercially cleaned Ross machine at a retail store, the numbers of aerobes recovered from deep tissues were similar with both treatments. When halved cuts were treated in one or other machine, deep tissue contamination was greater with the Lumar machine than with the Ross machine. Contamination of deep tissues as a result of tenderizing by piercing with thin blades can be minimized if the blades are designed to limit the number of bacteria carried into the meat and the microbiological condition of incised surface is well controlled. PMID- 15830674 TI - Comparison of methods for assessing reverse osmosis membrane treatment of shrimp process water. AB - Interest in reuse of process water from the food industry has reinforced the importance of controlling and monitoring the effectiveness and reliability of treatment systems regarding removal of organic matter and microorganisms. The ability of adenosine triphosphate bioluminescence, conductivity, turbidometry, absorbance, and multichannel fluorescence spectroscopy for indirectly monitoring the integrity of a reverse osmosis membrane when treating process water recovered from peeling in a shrimp processing line was evaluated. This study demonstrated that reverse osmosis was capable of removing bacteria (ca. 7 log CFU ml(-1)) to the levels required by the regulatory authorities for water recycling within the same food unit operation. Adenosine triphosphate and turbidometry showed a higher sensitivity for detecting compromising conditions at the treatment system (0.1% concentration of feed in permeate) and a better correlation with the aerobic count at lower levels than the other methods investigated. The sensitivity for assessing membrane integrity of conductivity and multichannel fluorescence was 1% of feed in permeate. Impact of feed variations was best leveled out in the permeates for turbidity measurements. Multichannel fluorescence spectroscopy may require laborious calibration procedures and expertise regarding data analysis and interpretation of results, which are not always available in food industries. Absorbance did not respond to changes in membrane integrity and was not well correlated to the aerobic count because of the poor sensitivity of this method for these purposes. PMID- 15830675 TI - A beta-glucuronidase-based agar medium for the differential detection of the yeast Debaryomyces hansenii from foods. AB - A selective and differential solid medium, Debaryomyces differential medium (DDM), was used for the isolation of Debaryomyces hansenii. This medium is formulated to allow detection of the beta-glucuronidase enzyme using the chromogenic substrate magenta-glucuro.CHA (5Br-6Cl-3indolyl-beta-D-glucuronide, cyclohexylammonium salt). Of the more than 120 microorganisms tested, including yeasts, bacteria, and a filamentous fungus, only D. hansenii produced violet colonies, thus permitting its easy discrimination from other organisms. When quality assessment tests were performed, optimal productivity and selectivity were obtained for D. hansenii. The medium was also satisfactory when used to test naturally contaminated food products. PMID- 15830676 TI - Identification of fungal contamination and determination of mycotoxigenic molds by micellar electrokinetic capillary chromatography in smoked paprika. AB - The purpose of this work was to analyze the fungal contamination in smoked and unsmoked paprika processed from different cultivars of pepper and to investigate the ability of these and other mycotoxigenic molds to grow and synthesize mycotoxins in smoked paprika. Eighteen mycotoxins were evaluated using micellar electrokinetic capillary chromatography. No relevant differences were found in fungal contamination between smoked and unsmoked paprika. The number of yeasts obtained was low, ranging from 0.4 to 3.29 log CFU g(-1); most of the yeast strains were identified as Cryptococcus spp. followed by Candida spp. All mold counts were <4 log CFU g(-1). Aspergillus, Cladosporium, Penicillium, and Fusarium were the predominant hyphomycete genera. Six mycotoxins were identified in the extracts of several strains isolated from paprika and incubated on malt extract agar. Penicillium expansum followed by Penicillium citrinum and Penicillium raistrickii were the dominant mycotoxigenic fungi isolated. Most of themycotoxin-producing fungi produced detectable amounts of mycotoxins when grown on paprika agar. PMID- 15830677 TI - Glial fibrillary acidic protein and myelin basic protein as markers for the immunochemical detection of bovine central nervous tissue in heat-treated meat products. AB - Because bovine central nervous tissue (CNT) is the main risk material in transmission of the infective agent of bovine spongiform encephalopathy, a suitable test is needed to enforce the ban on CNT in human foodstuffs in the United States and the European Union and to ensure that meat products are free of CNT To detect bovine CNT in heat-treated meat products, we used immunohistochemistry and Western blots with antibodies against glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP). Both antigens were resistant to processing methods used for meat products. The anti-GFAP antibody showed a high degree of tissue specificity, whereas the anti-MBP antibody had high species specificity, clearly differentiating between porcine and bovine CNT Therefore, immunochemistry performed with both proteins provides an effective means for detecting bovine CNT in meat products. PMID- 15830678 TI - Low detection efficiency of candling as a commonly recommended inspection method for nematode larvae in the flesh of pelagic fish. AB - The third-stage larvae of the parasitic nematode Anisakis simplex commonly occur in most commercially important fish species from the North Atlantic. In this study, the occurrence and site distribution of Anisakis larvae in various size groups of Norwegian spring spawning herring (Clupea harengus), blue whiting (Micromesistius poutassou), and mackerel (Scomber scombrus) from the Northeast Atlantic was investigated. Although most of the larvae were found in the abdominal cavity, the small proportion lodged in the flesh was sufficient to affect food quality and safety. However, the prevalence of Anisakis larvae in fish fillets varied considerably among the size groups of each species: 15 to 60% in herring, 32 to 77% in mackerel, and 89 to 100% in blue whiting. The commonly recommended nematode detection method in the fish processing industry is candling, i.e., a brief visual inspection on a light table. Comparison of successively more accurate detection methods, i.e., candling, enzymatic degradation, and UV illumination, for fillets of each fish species revealed that only 7 to 10% of the nematode larvae present in the fillets were detected by candling. The candling efficiency was apparently independent of fillet thickness. Thus, candling is not sufficient to detect the majority of the nematodes that are actually present in the fillets of pelagic fish from the Northeast Atlantic. These findings emphasize the importance of adequate deep-freezing of any fresh pelagic fish product, especially if it is intended for consumption in a raw or semiraw state. PMID- 15830679 TI - A real-time inspection system using a terahertz technique to detect microleak defects in the seal of flexible plastic packages. AB - A method to detect production faults in flexible plastic packages with the use of terahertz radiation is presented. Relying on the large difference between the absorption coefficients of plastic and water (for water-filled channel defects) and on the refraction index difference between plastic and air (for air-filled channel defects), our technique consists of focusing and scanning a terahertz beam on the sealed area of the package, followed by detection of the transmitted signal. Compared with previous methods, such as visual and ultrasound inspection, our technique can be applied to optically opaque packages and does not require immersion in a matching liquid. We tested our system on defects that we fabricated as water-filled and air-filled channels imbedded in polyethylene films, with diameters in the range of 10 to 100 microm. The detection limit (the minimum size of a detectable defect) depends on the conveying speed; this relationship was determined and analyzed. The results show that our system has the potential for application in an actual production line for real-time inspection. PMID- 15830680 TI - Survival of Shigella boydii 18 in bean salad. AB - A strain of Shigella boydii 18 involved in a bean salad outbreak and S. boydii 18 ATCC 35966 were used to inoculate bean salad. Bean salad samples were stored at 4 or 23 degrees C. At 4 degrees C, the S. boydii survived for the duration of the shelf life of the salad but did not grow. At 23 degrees C, both strains increased by two orders of magnitude by day 2 and decreased rapidly thereafter. This demonstrates the importance of proper storage in preventing the outgrowth of foodborne pathogens. PMID- 15830681 TI - Destruction of Salmonella Senftenberg on the shells of intact eggs by thermoultrasonication. AB - The combined effect of ultrasonic waves and heat treatment applied simultaneously was evaluated on the survival of strains of Salmonella Senftenberg on shells of intact eggs. This combined process has a higher killing effect than heat treatment alone. The decimal reduction times (D-values) were decreased by 65.2 to 11.1% in the temperature range studied (57.8 to 67 degrees C). In contrast to the effect on Salmonella enterica serovar Enteritidis in a previous study, thermoultrasonication had no important advantage for elimination of Salmonella Senftenberg. However, because 52 degrees C is a nonlethal temperature for Salmonella Senftenberg, the conditions used for the elimination of Salmonella Enteritidis (52 degrees C for 12 min) in the previous study would be equivalent to ultrasonic treatment alone in the present study. This thermoultrasonication treatment may result in a 100-fold greater reduction of Salmonella Senftenberg than that achieved by common in-shell egg pasteurization (60 degrees C for 3.5 min). PMID- 15830682 TI - Molecular analysis of Salmonella isolates recovered from processed Turkey carcasses. AB - This study was carried out to determine the prevalence of some virulence characteristics associated with Salmonella isolates recovered from processed turkey carcasses in the Midwestern region of the United States. A total of 94 Salmonella isolates recovered from turkey carcasses from two processing plants (A and B) were examined to determine the prevalence of invA, pagC, and spvC genes. Bioassays also were used to evaluate aerobactin and colicin production. All isolates (100%) were positive for the presence of invA and pagC but were negative for spvC. Overall, 19.1% of all isolates tested were positive for aerobactin production, and 25.5% of all isolates were positive for colicin. Aerobactin and colicin production differed among isolates recovered from the two plants; more isolates from plant B produced these compounds. The Salmonella isolates examined in this study possess significant potential for causing human illness. PMID- 15830683 TI - Prevalence of Yersinia enterocolitica in fattening pigs. AB - The prevalence of pathogenic Yersinia enterocolitica in pig herds was monitored during six trials (at four different farrow-to-finisher farms). Samples were taken throughout the whole rearing period from birth of the piglets to the final fattening stage, and different samples were taken from these pigs during the slaughter process. Environmental samples also were evaluated to identify potential sources of on-farm infection. Y. enterocolitica was isolated using irgasan-ticarcillin-potassium chlorate broth enrichment and cefsulodin-irgasan novobiocin agar culture. Colonies were identified using bio- and serotyping methods and by PCR assay. Pathogenic Y. enterocolitica were not isolated from fecal samples from piglets and weaners. The only fecal samples positive for Y. enterocolitica were obtained during the fattening stage. The prevalence of Y. enterocolitica in fattening pig herds ranged between 0 and 65.4%. Y. enterocolitica isolates were detected at the abattoir in 38.4% of the tonsils, in 3.8% of the ileocecal lymph nodes, on 0.3% of the carcass surfaces before chilling, and on 0% of the carcass surfaces after chilling. Almost all isolates belonged to bioserotype 4/O:3. Only one strain was identified as O:9. All isolates contained the ail gene. The yopT gene was found in 99.1% of the farm isolates but in only 76.6% of the isolates found at the abattoir from the corresponding carcasses. Although a direct link between porcine isolates and human infection has not been demonstrated, the similarity of the bioserotypes in infected pigs and humans and the presence of virulence factors in porcine isolates should encourage further studies to determine the risk of transmission of Y. enterocolitica to humans from pigs and pork products. PMID- 15830684 TI - Fate of Listeria monocytogenes inoculated onto the surface of model Turkey frankfurter pieces treated with zein coatings containing nisin, sodium diacetate, and sodium lactate at 4 degrees C. AB - The antimicrobial effects of zein coatings containing nisin, sodium lactate, and sodium diacetate against Listeria monocytogenes on turkey frankfurters at 4 degrees C were determined. Our objectives were to determine whether zein, nisin, lactate, and diacetate alone or in combination could control the growth of L. monocytogenes on full-fat turkey frankfurters at 4 degrees C and to determine whether lactate or diacetate had any synergistic effect on the activity of nisin. Turkey frankfurter pieces surface inoculated with L. monocytogenes strain V7 were treated with zein-ethanol-glycerol (ZEG), zein-propylene-glycol (ZPR), ethanol glycerol (EG), propylene glycol (PR), nisin (N), sodium lactate (L), or sodium diacetate (D) alone or in combination. Over 28 days, treatment with N or D alone reduced L. monocytogenes counts on frankfurters by 6.6 or 6.3 log CFU/g, respectively. N-D treatment reduced L. monocytogenes by 6 log CFU/g. The zein solvents EG and PR reduced L. monocytogenes by about 5.6 and 5.2 log CFU/g, respectively, similar to the results obtained with ZEG and ZPR, which suggests that zein powder per se had no antimicrobial activity. After 28 days, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, and ZPR-N-D-L yielded no detectable CFU. L alone was ineffective. No synergies were observed. N and D when used singly and the combinations of N-D, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, ZPR-N-D-L, EG, and PR were effective as inhibitors of the growth of recontaminating L. monocytogenes cells on full-fat turkey frankfurters. PMID- 15830685 TI - Spore formation in Bacillus subtilis biofilms. AB - Spore formation by a Bacillus strain (Bacillus subtilis SpoIVFB-GFP) engineered with a green fluorescent protein (GFP) fused to a polytopic membrane protein (SpoIVF) that fluoresces during sporulation was observed. Biofilms of B. subtilis SpoIVFB-GFP containing ca. 8 log CFU/ml vegetative cells and spores below the lower detection limit (i.e., <1 log CFU/ ml) were allowed to develop on glass wool (37 degrees C). These biofilms were subsequently exposed to nutrient limitation to stimulate spore formation, which was monitored for fluorescence by confocal scanning laser microscopy. Sporulation in corresponding planktonic cells was also monitored for comparative purposes. Planktonic B. subtilis SpoIVFB-GFP cells began fluorescing after 5 h, while B. subtilis SpoIVFB-GFP biofilm cells began fluorescing after 30 h. Results suggested that an existing biofilm of vegetative B. subtilis cells may be stimulated to form spores when exposed to conditions of nutrient limitation. From a practical point of view, it may be suggested that a window of time does exist before sporulation occurs in attached Bacillus biofilms highlighting the need for shorter operating runs between cleaning and sanitation of food-processing equipment surfaces. PMID- 15830686 TI - Effectiveness of trisodium phosphate treatment against pathogenic and spoilage bacteria on poultry during refrigerated storage. AB - To determine the efficacy of trisodium phosphate (TSP) against pathogenic and spoilage bacteria on poultry and to assess the influence of the bacterial combination of inoculum on TSP effect, chicken legs were coinoculated with similar concentrations of a pathogenic (Salmonella enterica ser. Enteritidis or Listeria monocytogenes) and a spoilage (Pseudomonas fluorescens or Brochothrix thermosphacta) bacteria. Samples were dipped in TSP (12%, 15 min) or were not treated (control). Microbiological analysis and pH determinations were carried out at 0, 1, 3, and 5 days of storage (3 degrees C). Significant bacterial reductions (marked in gram-negative species) were observed on TSP-treated samples throughout refrigerated storage. Inoculum composition scarcely influenced the TSP effect against gram-positive bacteria. However, greater reductions were observed on gram-negative bacteria (Salmonella Enteritidis and P. fluorescens) when samples were coinoculated with B. thermosphacta and L. monocytogenes, respectively. Values of pH were higher in TSP-treated than in control samples at all sampling times. The combination of bacteria in inocula did not have a significant influence on pH values. PMID- 15830687 TI - Use of fluorescent microspheres as a tool to investigate bacterial interactions with growing plants. AB - Foodborne pathogens may exist as endophytes of growing plants. The internalization of Escherichia coli O157:H7 or other foodborne pathogens in growing lettuce plants may be independent of microbial factors. Mature lettuce plants were surface irrigated with E. coli O157:H7 or with FluoSpheres (fluorescent microspheres) and harvested 1, 3, and 5 days post-exposure. FluoSpheres were utilized as a bacterial surrogate. Microscopic examination of root, stem, and leaf tissue sections revealed that FluoSpheres were internalized into growing plants. Laser scanning confocal microscopy revealed that FluoSpheres were present within the root tissue and leaf stem tissue. The presence of FluoSpheres in internal portions of stem and leaf tissue suggests transport of the spheres from the root upward into the edible tissue. The level of uptake of FluoSpheres and E. coli O157:H7 was quantified using filtration. Numbers of FluoSpheres and E. coli O157:H7 cells in plant tissue were similar. The entry of E. coli O157:H7 into lettuce plants may be a passive event because the concentration of FluoSpheres was similar to that of the pathogen. PMID- 15830688 TI - Multiplex PCR method for the simultaneous detection of histamine-, tyramine-, and putrescine-producing lactic acid bacteria in foods. AB - In a screening of primers, we have selected three pairs of primers for a multiplex PCR assay for the simultaneous detection of lactic acid bacteria (LAB) strains, which potentially produce histamine, tyramine, and putrescine on fermented foods. These primers were based on sequences from histidine, tyrosine, and ornithine decarboxylases from LAB. Under the optimized conditions, the assay yielded a 367-bp DNA fragment from histidine decarboxylases, a 924-bp fragment from tyrosine decarboxylases, and a 1,446-bp fragment from ornithine decarboxylases. When the DNAs of several target organisms were included in the same reaction, two or three corresponding amplicons of different sizes were observed. This assay was useful for the detection of amine-producing bacteria in control collection strains and in a LAB collection. No amplification was observed with DNA from nonproducing LAB strains. This article is the first describing a multiplex PCR approach for the simultaneous detection of potentially amine producing LAB in foods. It can be easily incorporated into the routine screening for the accurate selection of starter LAB and in food control laboratories. PMID- 15830689 TI - Analysis of mercury in sportfish tissue by thermal decomposition, amalgamation/atomic absorption spectrophotometry. AB - Sportfish samples (n = 133) that originated from Indiana waters were analyzed for total mercury using inductively coupled plasma/mass spectrometry (ICP/MS) and thermal decomposition, amalgamation/atomic absorption spectrophotometry (TDA/ AAS). Total mercury concentrations obtained by the two methods were not significantly different (P > 0.05). The correlation coefficient for total mercury obtained for the two methods was 0.94. The limit of detection using TDA/AAS was 0.1 ppm. TDA/AAS is a preferred technique for the analysis of total mercury in fish tissue because it is rapid (6 min per sample) and easy to use and requires little sample preparation. PMID- 15830690 TI - Brain tissue fragments in jugular vein blood of cattle stunned by use of penetrating or nonpenetrating captive bolt guns. AB - Although the incidence of bovine spongiform encephalopathy in cattle continues to decline in the United Kingdom, it remains important to maintain vigilance of all potential routes of transmission of infection to humans. Initial studies have demonstrated a potential risk of carcass contamination with brain tissue following the use of captive bolt gun stunning in cattle. The objective of this study was to further explore these initial findings particularly in regard to captive bolt guns currently in use in the United Kingdom. Brain tissue fragments or elevated levels of a marker protein for brain tissue were detected in venous blood samples from 4% (95% confidence interval, 1.6 to 9.8%) of cattle stunned by penetrating captive bolt gun and from 2% (95% confidence interval, 0.6 to 7%) of those stunned by nonpenetrating captive bolt gun. PMID- 15830691 TI - [Application of the tragus cartilage ring with perichondrium in open-ended tympanoplasty]. AB - OBJECTIVE: To investigate the effect of the application of tragus cartilage ring with perichondrium in open-ended tympanoplasty and it's long-term results. METHOD: The tympanic membrane perforations were treated by the surgery of open ended tympanoplasty. The tragus cartilage ring with perichondrium autograft in 38 patients with otitis media were adopted in the operation from 1995 to 2000. After operation,the patients were followed up in 2 weeks, 1 month, 3 months, 6 months, 1 year and 3 years. RESULT: Of 38 cases, 4 failed but the othershealed in one stage. The successful rate was 89.47%. After 2 weeks, among the 4 cases,2 had grafts crannies and 2 cases were infected. But when they accepted the second operation and after a following up of three years, the successful rate was 100%. Grafts were atrophy in 2 cases after 6 months to 3 years. Before the operation the average air conduction hearing was (45.66+/-8.40)dB HL and the difference between bone and air was less to (26.05+/-8.15)dB HL. From 1 months to 3 years after the operation, the air conduction and bone conduction ware respectively (31.58+/-7.45)dB HL and (19.61+/-6.41)dB HL in 1 month, (26.18+/-7.02)dB HL and (12.63+/-7.59)dB HL in 3 months,(23.55+/-7.70)dB HL and (10.79+/-5.52)dB HL in 6 months,and (23.42+/-6.99)dB HL and (11.05+/-5.95)dB HL in 1 years, keeping at limits of (23.95+/-6.69)dB HL and 10.53+/-5.43 dB HL in 3 years. There were significant differences between pre-operation and post-operation in 1 month,and 3 months. But there were no significant differences after operation in 6 months, 1 year, 3 years and 3 months. CONCLUSION: The tragus cartilage ring with perichondrium is of great importance to increase the guality of tympanoplasty and effective rate owing to its reliable technique and material collection. PMID- 15830692 TI - [Expression of hypoxia-inducible factor-1alpha in middle ear cholesteatoma]. AB - OBJECTIVE: To explore the pathogenetic mechanism of middle ear cholesteatoma, the aim of this study is to detect the expression of Hypoxia-inducible factor-1alpha in middle ear cholesteatoma and normal external ear canal skin. METHOD: We used the technology of immunohistochemistry to examine the expression of Hypoxia inducible factor-1alpha in thirty-one middle ear cholesteatomas and ten samples of normal external ear canal skin. RESULT: The expression of HIF-1alpha was extremely higher in middle ear cholesteatomas than in normal external ear canal skin (P <0.05). CONCLUSION: We found that the higher expression of Hypoxia inducible factor-1alpha in middle ear cholesteatomas, so we think that Hypoxia inducible factor-1alpha play an important role in the pathogenetic process of middle ear cholesteatoma, and hypoxia may be an incentive in the pathogenetic mechanism of middle ear cholesteatoma. PMID- 15830693 TI - [Analysis of preoperative findings and ossicular condition in chronic suppurative otitis media]. AB - OBJECTIVE: To discussion the relationship of preoperative findings and ossicular condition in chronic suppurative otitis media. METHOD: The correlation between the ossicular conditions and classification of tympanic membrane perforation, ear discharge, air conduction pure tone average, air-bone gap, pneumatization, complication, and cholesteatoma in 251 patients(288 ears) with chronic suppurative otitis media was analysed. RESULT: The air-conduction threshold and air-bone gap in patients with ossicular discontinuity are higher than that in patients with ossicular continuity. Ossicular discontinuity in patients with perforation of the pars flaccida of tympanic membrane, persistently draining ears, complications, and cholesteatoma occurred significiantly more frequently than those without these conditions. CONCLUSION: The air-conduction threshold and air-bone gap are the more reliable indications to identify the ossicular conditions in patients with chronic suppurative otitis media. There are significiant correlation between the ossicular conditions in patients with chronic otitis media and their classification of tympanic membrane perforation, ear discharge, complication, and cholesteatoma. PMID- 15830694 TI - [Surgical experience by 95 malformed stampedes and otosclerosis]. AB - OBJECTIVE: To investigate the effect of stapes surgery for conductive hearing loss with intact tympanic membrane and eustachian tube. METHOD: Stapes surgery data of 95 conductive hearing loss patients, which with intact tympanic membrane and eustachian tube, was retrospective analyzed,and the surgical effect of otosclerosis, middle ear deformities and external canal stenosis was summarized. RESULT: Postoperative (4 weeks after the surgery) pure tone threshold test demonstrated that the effect of Vestibulotomy was better than incus-footplate connection (incus and footplate is connected by a cortical bone ) ( P <0.05). The surgery to the patients with external canal stenosis was better than those with otosclerosis or middle ear deformities ( P <0.05). There was no statistic significance between physiological stapedectomy and Vestibulotomy. The follow-up of 1-3 years showed that the pure tone threshold of 8 out of 14 cases reverted to the preoperative level, while the hearing improvement in the other cases still remained. CONCLUSION: Stapes surgery to the patients with stapes-oval window deformities is of significance compared with those with otosclerosis. PMID- 15830695 TI - [Congenital external and middle ear malformation-with 62 ears report]. AB - OBJECTIVE: To determine the relationship between external ear malformation and middle ear abnormality, and also the results of surgical treatment. METHOD: Fifty two cases with external ear malformation (62 ears) was dividied into two groups:the minor group and severe group. All the patients received surgical treatment, 48 ears by transmastoid approach and 14 ears by "direct entrance" approach. RESULT: The middle ear abnormality was minor in minor group,and severe in the patients with severe external ear malformation. In 93.5% cases the hearing threshold improved more than 15 dB in 3 months after operation, and reduce to 85.5% in 3 years follow up. CONCLUSION: The middle ear malformation is corresponding to external ear malformation. The effect of surgical treatment is different according to the degree of malformation and surgical complication. PMID- 15830696 TI - [Tinnitus with normal hearing and evoked otoacoustic emissions]. AB - OBJECTIVE: To investigate the objective methods for tinnitus exploration by analysing the cochlear activity in tinnitus patients with normal hearing. METHOD: The 50 cases (50 ears) of unilateral tinnitus with normal hearing (the plastic group 1), 24 cases (46 ears) of bilateral tinnitus with normal hearing(the plastic group 2) and the 34 persons (68 ears) of normal hearing without tinnitus (control group) were measured by evoked otoacoustic emissions (TEOAE and DPOAE). All the tinnitus patients were also tested by the subjective tinnitus frequency and analysed with appropriate characteristic in TEOAE and DPOAE audiogram. RESULT: (1) The passing rate of TEOAE was 64.0% in the tinnitus group 1, 91.3% in the group 2, and 100% in the control group (group 1 P < 0.01, group 2 P >0.05 ). (2) The passing rate of DPOAE was 100% in three groups. But comparing to the control group, the amplitudes of DPOAE at the limited frequency ranges (3.125 kHz and 8.837 kHz) of the tinnitus group 1 was significant decreased ( P <0.05). (3) The subjective tinnitus frequencies were 0.25 approximately 8.00 kHz. The decreases in amplitudes and absent within the tinnitus frequencies were observed in TEOAE and DPOAE audiogram. CONCLUSION: Some patients of normal hearing with tinnitus have had early harmness of outer hair cell in cochlear. Evoked otoacoustic emissions is one of worthy methods to evaluated objectively the tinnitus. TEOAE is more susceptible to harmness in cochlear than DPOAE. DPOAE is superior to the analysis of tinnitus frequencies. PMID- 15830697 TI - [Application of extended high frequency audiometry in the patients with chronic renal failure treated by hemodialysis]. AB - OBJECTIVE: To explore the features of hearing changes in patients with renal insufficiency treated with hemodialysis. METHOD: Forty-nine patients with chronic renal failure ongoing hemodialysis and 66 normal controllers were enrolled in this study. RESULT: The changes of the averaged hearing thresholds of high frequency for hemodialysis subgroup were similar to those of the control group, but the impairment of the hearing was more severe. Compared with the control group, there was significant statistically differences in every age segments. CONCLUSION: The hearing impairment in patients with chronic renal failure treated with hemodialysis is characteristic by decreased in high frequency portion. PMID- 15830698 TI - [Auditory neuropathy in children(analysis of 14 cases)]. AB - OBJECTIVE: To study the clinic findings and the audiological characteristics of auditory neuropathy in children. METHOD: Clinical findings and systemic audiometry were analyzed in 14 cases(28 ears) of auditory neuropathy in children. RESULT: In 14 cases the primary onset age was 7 approximately to 13 year-old,4 male and 10 female. 7 cases had differential pitch tinnitus,mainly low pitch with poor speech discrimination. 1 case manifested peripheral neuropathy symptoms at 1 year after diagnosis. Audiograms of pure tone threshold (PTT) showed various curves including ascending slope, spoon, peak, lain letter S, letter W, gentle slope. Both ascending slope and spoon were 53.57% (15/28 ears). PTT revealed a elevated range from mild to moderate degrees. There was no significant difference between the cours for 0.5 approximately 1.0 year and 1.5 approximately 2.0 years ( t =0.24, P >0.05). Either the acoustic reflex (AR) was absent or the AR threshold was elevated. Speech discrimination scores (SDS) were 0% approximately 46%, average (14.64+/-15.42) %. Auditory brainstem response (ABR) was absent or severely abnormal. Prevalence of distortion product otoacoustic emissions (DPOAE) were 100%, amplitude of DPOAE was normal, which could not be suppressed by contralateral white noise. Prevalence of spontaneous otoacoustic emissions (SOAE) were 82.14% (23/28 ears). CONCLUSION: Audiological characteristics in pediatric auditory neuropathy is obviously. Systemic audiometry has important clinical signification in recognition, diagnosis and research of auditory neuropathy. PMID- 15830699 TI - [The value of grommet insertion for middle ear effusion in the patients of nasopharyngeal carcinoma after radiotherapy]. AB - OBJECTIVE: To evaluate the value of grommet insertion for middle ear effusion in the patients of nasopharyngeal carcinoma after radiotherapy. METHOD: Thirty-two recrudescent cases(51 ears) with middle ear effusion after myringotomies were treated by grommet insertion again,and then,the eustachian tube function,hearing and the other symptoms were observed, the effect were evaluated. RESULT: 15.7%(8/51) ears were cured,the total efficiency rate is 96.1%(49/51). The eustachian tube function had been straightened up in 13.7%(7/51)ears,the other symptoms were improved in different degrees for the cars of setting grommet, 25.5% ears occurred otorrhea, 9.8% ears occurred perforation. CONCLUSION: Grommet insertion is a favorable way for the eustachian tube function straightened up. For the ears that the eustachian tube function damaged seriously, repeated grommet insertion is a good measure for keeping the ear function. In a word, the benefit is more than the loss after grommet insertion. PMID- 15830700 TI - [Pharmacokinetics of the methylprednisolone, dexamethasone and hydrocortisone in the inner ear after administered via round window]. AB - OBJECTIVE: To compare the pharmacokinetics of Methylprednisolone, Dexamethasone and Hydrocortisone in the inner era of guinea pig after administered via round window. METHOD: Guinea pig was divided into groups chronologically. After a drop of 100 g/L corticosteroid applied into the round window niche, the tympanic perilymph was sampled chronologically. Concentration of the three corticosteroid in perilymph were measured via HPLC and analyzed by 3P97 software. RESULT: The highest concentration of the three drug appeared at 30 min, 60 min and 30 min. Then the concentration declined rapidly. CONCLUSION: All three corticosteroid permeate the round window membrane easily, but the concentration declined rapidly after the peak appearing. PMID- 15830701 TI - [Observation on the protective effect of MK-801 against hearing loss in acoustic trauma]. AB - OBJECTIVE: To observe the protective effects of MK-801, an antagonist of N-methyl D-aspartate (NMDA) receptor, against temporary (TTS) or permanent threshold shifts (PTS) in acoustic trauma. METHOD: Twenty guinea pigs were exposed to octave band noise with a central frequency of 4 kHz, 110 dB SPL for 3 h, and another twenty guinea pigs, to the same band noise at 115 dB SPL for 5 h. The test animals were given MK-801 (0.5 mg/kg, i. p. ) before and immediately after noise exposure; the control animals were treated with saline solution. Auditory brainstem responses (ABRs) of animals were recorded and ultrastructure of inner hair cells and afferent synapses were examined with transmission electron microscopy. RESULT: There was no significant difference in temporary threshold shifts immediately after noise exposure to 110 dB noise between the test and control group. The threshold shifts disappeared in one week after exposure in both groups. In 115 dB group, the threshold shifts of the test animals 7 days after exposure were significantly less than those of the control animals ( P >0.05) and 3 weeks after exposure, there was still this difference. The inner hair cell and afferent dendrites of the control animal exposed to 115 dB noise displayed many vacuoles. No vacuoles, however, were found in the inner hair cells or in the afferent dendrites of animals that were exposed to 110 dB noise or the test animals of 115dB group that received MK-801. CONCLUSION: MK-801 could partially protects against PTS, but not TTS in acoustic trauma through preventing from vacuole degeneration in inner hair cell and afferent dendrites. PMID- 15830702 TI - [Rehabilitation therapy after nasal endoscopic surgery for nasal polyp and chronic sinusitis patients]. AB - OBJECTIVE: To analyze the possible factors affecting the results of nasal endoscopic surgery in nasal polyp and chronic sinusitis patients and to discuss the ways to improve the results of operation. METHOD: Retrospective analysis was done in 226 patients followed up more than 3 year. The analysis was focused on patients' occupation, personal habits, extent of persisting on rehabilitation therapy after operation and clinical recovery status. RESULT: The patients who work in the environment rich of irritating substance and dust,the patients having unhealthy habits (e. g: spicy food and alcohol addicting) and the patients who did not take or insist on rehabilitation therapy after operation (e. g: local and systemic medication, nasal lavage and cleaning up of surgical cavities under nasal endoscope) had higher frequency of relapse and quicker recurrence. Rehabilitation therapy was the most important factor affecting the results of operation. CONCLUSION: Insisting on systemic rehabilitation therapy after operation is a occupation protection and kicking the unhealthy personal habits can prevent and reduce the recurrence of nasal polyp and chronic sinusitis after nasal endoscopic surgery. PMID- 15830703 TI - Potential benefits of co-transplanting autologous adult stem cells together with human embryonic stem cells or their differentiated derivatives. PMID- 15830704 TI - Configuration of a new bioartificial liver support system and in vitro evaluation of its functions. AB - The aim of this study was to configure a new bioartificial liver (BAL) support system and evaluate its functions in vitro. Chinese experimental miniature pig hepatocytes were isolated by an in situ recirculating collagenase perfusion method and were cultured in serum-free medium with restriction attachment and spinner technique to form hepatocyte spheroid suspensions containing 1.0 x 10(10) hepatocytes. The BAL support system was configured by inoculating the hepatocyte spheroids into the cell circuit of a hollow fiber bioreactor (BIOLIV A3A, Cell Biotech Limited, HK, China). The number and viability of hepatocytes, the levels of alanine aminotransferase (ALT), total bilirubin (TBi), and albumin (ALB) in the circulating hepatocyte suspension and RPMI-1640 medium, and lidocaine metabolism were determined during 6 hr of circulation in the BAL devices. Independent experiments were performed 5 times. There were no significant changes in the number and viability of the hepatocytes during the circulation period. The BAL support system demonstrated substantial albumin synthesis and lidocaine metabolism. The results indicate that the new BAL support system has the ability to perform liver functions and could be used to treat liver failure or provide temporary liver support in patients who are candidates for liver transplantation. PMID- 15830705 TI - Intracellular inhibitory effects of Velcade correlate with morphoproteomic expression of phosphorylated-nuclear factor-kappaB and p53 in breast cancer cell lines. AB - Velcade, a proteasome inhibitor, has been shown to inhibit DNA binding activity of nuclear factor-kappaB (NF-kappaB) and to stabilize p53 in vitro. But its impact, in the context of activated (phosphorylated and translocated) NF-kappaB and the expression of p53, has not been studied in breast cancer. It would be desirable to determine whether or not the immunohistochemical (IHC) expressions of activated NF-kappaB and of p53 can predict the effects of Velcade in viable tumor cells. To answer these questions, we selected 3 breast cancer cell lines (SKBR-3, MDA-175, and MDA-231), which are negative for hormonal receptors, but differ in HER-2/neu expression (strong, mild, and minimal, respectively). The 3 cell lines showed different expressions of phosphorylated (p)- NF-kappaB and p53, as evaluated using immunohistochemistry with visual quantification by brightfield microscopy. After being treated with Velcade for 2 days, MDA-231 cells showed markedly reduced proliferation, followed by SKBR-3 cells, and then by MDA-175 cells. There was strong correlation between the nuclear expression of either p-NF kappaB or p53 and the inhibitory rate of Velcade in the 3 cell lines (r = 0.987 and 0.807, respectively). Western blotting showed an increase in inhibitor-kappaB (I-kappaB) expression in nuclei of MDA-231 and SKBR-3 cells, but not in MDA-175 cells, following exposure to Velcade. Velcade treatment resulted in cleaved caspase-3 expression in MDA-231 cells and in the overexpression of p53 and p21WAF1 in all 3 cell lines, as evaluated using Western blotting. In summary, morphoproteomic analysis of p-NF-kappaB and p53 can be correlated with the inhibitory effect of Velcade in vitro. We propose that this proliferative inhibition is variably associated with blocking p-NF-kappaB function by upregulation of nuclear I-kappaB, stabilization of p53, and induction of p21WAF1. PMID- 15830706 TI - Application of real-time PCR and melting curve analysis in rapid detection of Ael and Bel blood types. AB - The ABO blood group is the most important blood group system in transfusion medicine. In addition to the normal levels of ABO antigen expression, Ael and Bel represent the two major blood types that have a weak expression of the A or B antigens on red blood cells. Due to the fact that typing of Ael and Bel by conventional serological methods is time consuming and sometimes gives false positive and false-negative results, it is warranted to develop an additional technique for the identification of Ael and Bel individuals. Through genetic analysis we have previously identified Ael as possessing an A allele with IVS6+5G ->A mutation (Transfusion 2003;43:1138-1144) and Bel as possessing a B gene with 502C-->T mutation in Taiwan (Vox Sanguinis 2003;85:216-220). Hence, real-time PCR based genotyping methods were developed in this study to facilitate the detection of Ael and Bel. For genotyping of Ael and Bel, the region of mutations was PCR amplified and subjected to the LightCycler (LC) real-time PCR assay using LC Red640-labeled hybridization probe. Melting curve analysis was performed to determine the melting temperature Tm that was used for genotype detection of Ael and Bel blood types. For Ael genotyping, the melting curve of the normal control appears as one peak at 59.19+/-0.07 degrees C (mean+/-SE) and that of Ael appears as 2 peaks at 59.21+/-0.07 degrees C and 64.39+/-0.07 degrees C, corresponding to the O and Ael alleles, respectively. For Bel genotyping, the melting curve of the normal control appears as one peak at 67.99+/-0.11 degrees C and that of Bel appears as 2 peaks at 59.99+/-0.12 degrees C and 68.1+/-0.13 degrees C, corresponding to the Bel and O alleles, respectively. This genotyping method was shown to be accurate, based on automated sequencing of the PCR-amplified products. It takes only 90 min to perform this genotyping test. Detecting the Ael and Bel blood types by combined LC-PCR and melting-curve analysis is a rapid, reliable, and easy method. PMID- 15830707 TI - Evaluation of an automated hematologyanalyzer (CELL-DYN 4000) for counting CD4+ T helper cells at low concentrations. AB - In human immunodeficiency virus (HIV) infection, CD4 cell counts are useful in defining the disease state, monitoring antiviral treatment, and identifying patients at risk for opportunistic infections. Counting CD4 cells typically relies on traditional immuno-flow cytometric analyzers that require opening the tube for manipulation of the blood sample. In addition to automated blood cell counting, the CELL-DYN 4000 hematology analyzer performs a completely enclosed and automated analysis of the T lymphocyte subsets. We studied the performance characteristics of this method in blood samples containing low levels of CD4+ T cells. In one set of experiments, we emulated low level CD4 counts by use of a CD4 Positive Isolation Kit to deplete the CD4+ cells from blood samples. We used the FACScan analyzer for reference counts. Measurements were made exactly 12 hr after venepuncture in samples that were stored at room temperature. In normal samples and those with low CD4+ cell counts, there was excellent correlation between the results of the CELL-DYN and FACScan methods. Using the CELL-DYN 4000 analyzer, the precision of CD4+ and CD8+ T-cell counts was high (CV = 2 to 8%). The CD4+ T-cell count was linear over a wide range (35 to 1640 cells/microl). This study shows that CD4 and CD8 T cell counts using the CELL-DYN 4000 analyzer is suitable for normal samples and also for those with low CD4+ T cell counts. The method is rapid and automated, and blood specimens remain enclosed, minimizing the biohazard of exposure to blood of HIV patients. PMID- 15830708 TI - Free radical scavenging, DNA protection, and inhibition of lipid peroxidation mediated by uric acid. AB - Uric acid (UA) has been proposed to be the dominant antioxidant in birds. The objective of this study was to investigate the quenching effect of varying concentrations of UA, including those found in avian plasma, on specific reactive oxygen species (ROS) and to determine the ability of UA to protect DNA and cellular membranes from ROS-mediated damage. Hydroxyl (OH) and superoxide (O2-) radicals were detected by electron spin resonance (ESR) and their presence was reduced following addition of UA (p <0.05) in a concentration-dependent manner. UA inhibited hydroxyl-mediated DNA damage, indicated by the presence of more precise, dense bands of lambda Hind III DNA after agarose gel electrophoresis and ethidium bromide staining (p <0.05). Lipid peroxidation of silica-exposed RAW 264.7 cell membranes was diminished (p <0.02) after addition of UA to the cell incubation mixture. These studies demonstrate that UA scavenges hydroxyl and superoxide radicals and protects against DNA damage and lipid peroxidation. These results indicate specific antioxidant protection that UA may afford birds against ROS-mediated damage. PMID- 15830709 TI - Predicting clinical outcomes in peritoneal dialysis patients using small solute modeling. AB - The power of published models of dialysis adequacy to predict clinical outcomes in renal failure patients maintained on continuous ambulatory peritoneal dialysis (CAPD) is controversial. Inflammation may be an important predictor of morbidity and mortality in CAPD. Baseline data from a 2-yr prospective, longitudinal study of peritoneal dialysis adequacy were analysed. Baseline measures of dialysis adequacy (urea clearance [Kt/V], efficiency number [EN], dialysis index [DI], dialysate-plasma creatinine ratio [D/ Pcreat], creatinine clearance [CrCl weekly PD]) as predictors of outcome were investigated by univariate analysis and by multiple logistic regression modelling. Baseline nutritional and inflammatory markers were also tested as predictors of outcomes. Outcomes were patient survival and technique failure over the succeeding 2 yr. Fifty-three patients consented to the study; 7 patients were unsuitable. Only 6 patients completed the study (13%). Non-survivors (n = 6) had lower protein catabolic rates and lower serum albumin concentrations, and higher C-reactive protein (CRP) levels at baseline than the patients who survived (p <0.05), but there were no differences in any of the measures of dialysis adequacy. The patient group that developed technique failure (n = 9) had significantly higher D/Pcreat (p = 0.037) at baseline. Serum albumin and CRP at study entry were significant negative and positive predictors of death respectively (p <0.05). No baseline variable achieved significance as a predictor of technique failure in the patient cohort. In conclusion, dialysis dose descriptors are poor predictors of clinical outcomes in CAPD patients. Inflammatory and nutritional markers such as CRP and albumin may be more important in predicting patient outcomes than measures of peritoneal small solute clearance. PMID- 15830711 TI - Albumin cobalt binding assay to rule out acute coronary syndrome. AB - The purpose of this study was to validate the Albumin Cobalt Binding (ACB) assay at the Seattle Veterans Affairs (VA) Hospital to determine if it would provide an earlier rule-out of acute coronary syndrome (ACS) in patients, compared to current use of cardiac injury markers. This study compares the distribution of ischemia modified albumin (IMA) values of our patient population to those provided by the kit manufacturer. IMA values were determined photometrically on a Roche Modular Analytical System on 200 subjects: 69 subjects not experiencing chest pain (normals), 78 subjects presenting to the emergency room (ER) with chest pain whose initial and subsequent troponin results were negative (non converters), and 53 subjects presenting to the ER with chest pain whose initial troponin result was negative but subsequent troponin results were positive (converters). Based on the relationships between IMA values in the initial samples from the non-converters and converters, we constructed a ROC curve to identify an optimum IMA rule-out value. The IMA values (mean+/-SD) for the normals, non-converters, and converters were 89+/-7.1, 100+/-13.9, and 126+/-14.1 U/ml, respectively, and each mean was statistically different from the means of the other groups. The ROC curve comparing converters and non-converters showed an area of 0.89 (p <0.001) compared to the line of identity. An IMA cut-off of 97 U/ml gives a 98% sensitivity and 45% specificity and may be the best decision point to differentiate between these groups in our population. Nine of 78 non converters were classified as having unstable angina. In conclusion, the ACB assay has a strong negative predictive value and sensitivity in our population for predicting the troponin results at 6 to 24 hr post-presentation. Because ACB results may be facility- and instrument-dependent, each facility should conduct an independent ROC analysis to determine the optimal IMA rule-out level. The ACB assay, when used in conjunction with cardiac injury markers and assessment of unstable angina, holds promise in reducing inappropriate low-risk hospital admissions and improving the clinical management of patients with chest pain. PMID- 15830710 TI - Progression of renal damage in the obese Zucker rat in response to deoxycorticosterone acetate-salt-induced hypertension. AB - This study assessed the progression of renal damage in obese Zucker rats in response to deoxycorticosterone acetate (DOCA)-salt-induced hypertension. Renal damage was evaluated by light microscopy and urine analysis at weekly intervals during the developmental phase of DOCA-salt hypertension and once during the plateau phase 42 days after the onset of treatment. Decreased tubular function was evident by day 8, as indicated by a significant increase in urine N-acetyl beta-D-glucosaminidase activity and glucose excretion. The tubular index, a measure of tubular damage, was significantly elevated by day 15 and continued to increase throughout the experiment. Glomerular damage, which was evident by day 8, was followed by increased urine albumin excretion by day 15. Only a few sclerotic renal glomeruli were apparent before the plateau phase; however, by day 42, approximately 50% of the glomeruli were sclerotic. Hyperplastic vascular changes were mild at day 8 and slowly increased in severity during the developmental phase. By day 42 the vascular changes were severe with some vessels so hyperplastic that their lumens were almost occluded. These findings show progressive changes in renal structure and function that begin as early as day 8 and increase progressively until severe changes are present at day 42, resulting in an end-stage PMID- 15830712 TI - Swimming exercise training prior to acute myocardial infarction attenuates left ventricular remodeling and improves left ventricular function in rats. AB - The effect of exercise training prior to acute myocardial infarction (AMI) on left ventricular (LV) remodeling is poorly understood. This study investigated the protective effect of 3 weeks of swimming exercise training prior to AMI on cardiac morphology and function. Male Sprague-Dawley rats (n = 35) were randomly assigned to 3 groups: swimming training (n = 14, 90 min, 5 days/wk, 3 wk), sedentary (n =14), and controls (n = 7, no exercise, no MI). At the end of the training/sedentary period, rats were subjected to AMI (ExMI and SedMI) induced by surgical ligation of the left coronary artery. Thereafter, the rats remained sedentary for a 4-wk recovery period. Trans-thoracic echocardiography was performed in each group at the end of the exercise/sedentary period (pre-AMI), 24 hr after AMI, and following recovery (4 wk after AMI). No differences were observed in LV dimensions and function pre-AMI among the 3 groups; however, LV end systolic diameter (LVESD) and LV-end systolic area (LVES-area) were significantly lower in the prior trained rats, 24 hr post-AMI with no additional change 4 wk post-AMI, during remodeling. Both LV-shortening fraction (SF%) and fractional area change (FAC%) were higher in the trained animals 4 wk post-AMI (39+/-12% vs 23+/-8%; p 0.002, and 48+/-14% vs. 38+/-9%; p 0.07, respectively). In conclusion, 3 wk of swimming exercise training prior to AMI significantly attenuated LV remodeling and improved LV function, despite no changes in LV dimensions or systolic function at the end of the exercise session. The data suggest that even a short-term training period is sufficient to induce cardiac protection. PMID- 15830713 TI - The influence of subarachnoid hemorrhage on neurons: an animal model. AB - Subarachnoid hemorrhage (SAH) has considerable mortality and morbidity, but the pathophysiologic mechanism is not entirely clear. Following SAH, blood or its lysate enters the subarachnoid space. This study examined how blood lysate influences the vulnerable brain following SAH. Heparinized hemolysate was slowly injected into the cisterna magna of 10 female rabbits, while a control group of 10 rabbits received a similar injection of heparinized isotonic sodium chloride solution without hemolysate. The basilar artery and brain tissue were excised after perfusion fixation. The degree of cerebral vasospasm was evaluated by measuring the cross-sectional area of the basilar artery, and brain damage was investigated by TUNEL staining. In the SAH group, the apoptosis index of neuronal cells located at the base of the temporal lobe averaged 26% (range = 3 to 56%), which was significantly higher than the corresponding apoptosis index in the control group (mean 0.5%, range = 0 to 4%, p <0.001). The mean cross-sectional area of the basilar artery in the SAH group did not differ significantly from that in the control group. These results suggest that SAH induces apoptosis of neuronal cells by a mechanism that is independent of cerebral vasospasm. PMID- 15830714 TI - Case report: subcutaneous elemental mercury injection--clinical observations and implications for tissue disposal from the histopathology laboratory. AB - A 31-yr-old man presented to his general physician complaining of pain and swelling in the soft tissue of both hips. He initially denied knowledge of the etiology, but after elemental mercury droplets were expressed from the skin wounds by manual manipulation, he admitted that the source was self-administered sc injections. He did not provide a reason for this or give the time-frame of the injections. The areas of skin discoloration and soft tissue induration were completely excised and the wounds healed uneventfully. Blood levels of mercury declined gradually after excision. Examination of the resected skin and subcutis revealed subcutaneous abscesses containing droplets of elemental mercury, released easily when the abscesses were sectioned. Sections showed dark droplets of metallic mercury associated with necrosis and microabscess formation, with areas of foreign body giant cell reaction. Disposal of the residual tissues and mercury by incineration would release the volatile elemental mercury into the atmosphere, where it would subsequently be returned to earth in rain water, be converted to methyl mercury by microorganisms, and enter the food chain. The US Environmental Protection Agency has recommended caution in consuming foods containing methyl mercury and has provided guidelines for consuming foods believed to contain increased amounts of mercury. Mercury is readily available in the United States by over-the-counter sales to consumers, and in some cultures elemental mercury is used in ritualistic practices. Skin injection cases are infrequent, but histopathology laboratories should recognize this phenomenon and be prepared to dispose of Hg-contaminated tissues in an environmentally sound manner. PMID- 15830715 TI - Immunohistochemical expression of Bax and Bcl-2 in penile carcinoma. AB - There is a complex interplay between the pro-apoptotic Bax and anti-apoptotic Bcl 2 family of proteins and the tumor suppressor gene p53. The pathogenic role of Bax and Bcl-2 protein expression in penile carcinomas has not previously been investigated. We examined Bax and Bcl-2 expression in verrucous (VC) and squamous cell carcinoma (SCC) of the penis. Herein we also present a concise review of p53, Bcl-2/Bax ratios, and their relationship to apoptosis. Fourteen cases of penile carcinoma, including 7 VC and 7 well-differentiated SCC, were analyzed for Bax and Bcl-2 expression by immunohistochemical analysis of paraffin embedded archived tissues. The number of positively staining tumor cells was enumerated per 100 tumor cells within non-overlapping high power fields. The Bax immunoreactivity was similar in VC (19+/-3%) and well-differentiated SCC (15+/ 4%) (p = 0.69). The expression of Bcl-2 protein was significantly higher in well differentiated SCC (69+/-12%) compared to VC (36+/-14%) (p = 0.04). The mean Bcl 2/Bax ratio was significantly lower in VC (1.89) compared to well-differentiated SCC (4.6) (p = 0.05). These findings indicate that penile VC and SCC are immunophenotypically distinct. Bax expression is comparable in verrucous and low grade squamous cell carcinomas, but Bcl-2 expression of Bcl-2 is significantly higher in the squamous cell carcinomas. PMID- 15830716 TI - Case report: glomus tumor of the colon. AB - Glomus tumors are tumors of pericytic origin and are usually found in the distal extremities. Glomus tumors have rarely been reported in viscera. The authors report a glomus tumor of the colon that caused rectal bleeding in a 40-yr-old man and was biopsied and excised endoscopically. The histology and immunohistochemical profile of the tumor are described and the literature on visceral glomus tumors is reviewed. PMID- 15830717 TI - Case report: atypical presentation of anti-Rg(a). AB - The Rodgers (Rg(a)) antigen is a plasma protein that binds to the red blood cell (RBC) membrane. About 2 to 3% of the transfusion-recipient white population lacks the antigen and can produce anti-Rg(a) antibody. We report the case of a 70-yr old man who presented with a medical history of hairy cell leukemia and profound pancytopenia that required RBC and platelet (PLT) transfusions. The patient had received 2 units of RBCs and 4 PLT concentrate pools. He was typed as O Rh(D) positive, with positive reactions in all 3-screen cells using the polyethylene glycol (PEG) indirect antiglobulin test/IAT (anti-IgG). Three antibody identification panels were performed, which all proved to be negative. A direct antiglobulin test and an auto-control were run, which were also negative. Since further investigations were needed, the patient's blood sample was sent to a reference laboratory where anti-Rg(a) was identified. Since the percentage of antigen-positive cells in the red cell panel was low, crossmatch compatible units of RBCs were transfused with no discernible immediate or delayed transfusion reaction. This report should alert hospital transfusion service personnel to recognize that, although the panel cells are usually reliable for antibody identification purposes, they may not have the antigens that are present on the screening cells. PMID- 15830718 TI - A note from history: two pioneering chemists, three hundred years apart. PMID- 15830719 TI - Sustained release characteristics of tablets prepared with mixed matrix of sodium carrageenan and chitosan: effect of polymer weight ratio, dissolution medium, and drug type. AB - The interpolymeric complexation of carrageenan and chitosan was investigated for its effect on drug release from polymeric matrices in comparison to single polymers. For this purpose, matrices with carrageenan: chitosan (CG:CS) ratios of 100%, 75%, 50%, 25%, and 0% were prepared at 1:1 drug to polymer ratio. The effect of dissolution medium and drug type on drug release from the formulations was addressed. Two model drugs were utilized: diltiazem HCl (DZ) as a salt of a basic drug and diclofenac Na (DS) as a salt of an acidic drug. Three dissolution media were used: water, simulated gastric fluid (SGF), and simulated intestinal fluid (SIF). Some combinations of the two polymers showed remarkable sustained release effect on DZ in comparison to the single polymers in water and SGF. However, no apparent effect for the combination on DZ release was shown in SIF. The medium effect was explained by the necessity of chitosan ionization, which could be attained by the acidic SGF or microacidic environment created by the used acidic salt of DZ in water, but not in SIF. An interaction between the medium type and CG:CS ratio was also found. With DS, the polymer combinations had similar dissolution profiles to those of the single polymers in water and SIF, which was explained by the lack of chitosan ionization by the medium or the drug basic salt. The dissolution profiles could not be obtained in SGF, which was attributed to the conversion of DS into diclofenac free acid. The importance of chitosan ionization for its interaction with CG to have an effect on the release of DS was demonstrated by performing dissolution of SGF presoaked tablets of DS in SIF, which showed an effect of combining the two polymers on sustaining the drug release. PMID- 15830720 TI - A comparison of trehalose dihydrate and mannitol as stabilizing agents for dicalcium phosphate dihydrate based tablets. AB - This study investigated the possible utility of trehalose dihydrate (TD) as a tablet stabilizing agent. Acetylsalicylic acid was used as the model hydrolyzable drug and dicalcium phosphate dihydrate (DCPD) as the base excipient, because it is well documented that ASA/DCPD tablets are unstable during storage at low temperature and high relative humidity; DCPD is usually combined with mannitol in order to improve tablet stability. Tablets comprising DCPD, 10% ASA, and 0%, 10%, or 20% w/w of TD were prepared by direct compression and stored at 35 degrees C and 82.9% relative humidity for 6 months. Additionally, control tablets with DCPD and ASA, only, or with DCPD, ASA and 20% mannitol, were also evaluated. At predetermined time intervals, formulations were tested for drug content, mechanical, microstructural, and drug dissolution properties. Additionally, thermal analyses and ASA solution stability studies were carried out. Results reveal that both TD and mannitol significantly reduce degradation of ASA included in DCPD-based tablets, but neither effectively protects against the marked decline in tablet mechanical properties on aging. The ASA stabilization effects of TD and mannitol were also observed in solution, indicating an interaction between these sugars and ASA. PMID- 15830721 TI - In vitro and in vivo evaluation of a novel nonscrotal matrix-type transdermal delivery system of testosterone. AB - A matrix-type testosterone (TS) transdermal delivery system for nonscrotal skin was developed with DuroTak 87-2510 as an adhesive polymer. When 3% dodecylamine was used in combination with 10% span 80, the in vitro rat permeation rate increased from 2.29 microg/cm2/hr to 6.51 microg/cm2/ hr as the TS loading dose increased from 2% to 6%. The maximum flux of experimental patch was about 14-fold higher than that of Testoderm. Release kinetics of TS from the patches was proportional to t1/2 following the Higuchi equation, and the release rate of TS increased as TS loading dose increased. Also, a good linear relationship between the skin permeation rate and the release rate was observed, which implies that the release rate is the rate-limiting process of the skin permeation. In vivo study showed that the plasma concentration of TS promptly increased and reached the peak level within 3-6 hours of application of the experimental patch. Area under the curve (AUC0 approximately 48) and Cmax also linearly increased in a dose-dependent manner up to 6% of TS. These results demonstrate the feasibility of developing a nonscrotal matrix-type transdermal delivery system of TS. PMID- 15830722 TI - Characterization of the novel ophthalmic drug carrier Sophisen in two of its derivatives: 3A Ofteno and Modusik-A Ofteno. AB - Sophisen, a new ophthalmic drug carrier, was characterized using physicochemical and morphological criteria. Diclofenac belongs to a nonsteroidal anti inflammatory molecule group and its ophthalmic use avoids side effects produced by steroid drugs. Cyclosporine-A is a cyclic peptide used as an immunosuppressive when administrated systemically. Its application in ophthalmology has been reported, but it is a very poor soluble drug. Diclofenac sodium and Cyclosporine A were mixed with Sophisen to render two new ophthalmic solutions that were named 3A Ofteno and Modusik-A Ofteno, respectively. Based on transmission electron microscopy and dynamic light scattering studies, we concluded that Sophisen is a polydisperse solution with a molecular weight of 413 +/-122 kDa, whereas 3A Ofteno and Modusik-A Ofteno are monodisperse solutions with molecular weights of 169 +/- 44 and 153 +/- 10, respectively. Sophisen was shown to be a good carrier for diclofenac sodium as evaluated by passive diffusion through the cornea. A comparative study suggests that diclofenac applied as eye drops was better tolerated when associated with Sophisen. In addition, Modusik-A Ofteno, a new aqueous solution of Cyclosporine-A, improved tear production in patients with moderate or severe dry eye condition. PMID- 15830723 TI - Potential of albumin nanoparticles as carriers for interferon gamma. AB - Although interferon gamma (IFN-gamma has been extensively studied as a potent activator for macrophages and as a promising adjuvant in vaccines, its rapid biodegradation and clearance have severely limited its clinical efficacy. Our major objective in this work was to develop formulation conditions to get high association of the cytokine to albumin nanoparticles, without leading any conformational changes and subsequent loss of activity. To achieve this objective, two different formulations were prepared by either 1) incubation between the cytokine and the newly prepared nanoparticles (IFN-NPA) or 2) between the protein and IFN-gamma prior coacervation (IFN-NPB). Steady-state fluorescence emission spectra revealed that the environment of the tryptophan (Trp) residue was not affected by conditions of mechanical stress required for preparing nanoparticles. A bioassay for antiproliferative activity with Hela cells indicated that the cytokine, after their desorption from the surface of nanoparticles (IFN-NPA), fully retained its activity. It also indicated that the cytokine was principally associated with nanoparticles via electrostatic interactions and confirmed by desorption experiments carried out in media with different pH and ionic strength, with burst effect ranked in the order pH 5 > pH 7.4 > pH 8.5. Also, the adsorption of IFN-gamma onto these carriers was able to improve the priming effects of IFN-gamma on the nitric oxide production (NO) by RAW macrophages. On the contrary, when we incubated the cytokine with the albumin solution prior to the desolvation process for preparing nanoparticles (IFN-NPB), we obtained better encapsulation efficiencies (around 100%), but the cytokine was inactive: it was not detected by ELISA or bioassay in Hela cells and unable to stimulate NO production by macrophages. PMID- 15830724 TI - Hydroxyzine- and cetirizine-loaded liposomes: effect of duration of thin film hydration, freeze-thawing, and changing buffer pH on encapsulation and stability. AB - PURPOSE: To assess the effect of the duration of film hydration, freeze-thawing, and changing buffer pH on the extent of entrapment of hydroxyzine and cetirizine, H1-antihistamines with different polarity, into liposomes, and the stability of these liposomes. METHODS: Multilamellar vesicles (MLV) were prepared by thin lipid film hydration using L-alpha-phosphatidylcholine (PC) and buffer containing 80 mg hydroxyzine at pH 7. For MLV containing hydroxyzine, the liposomes were subjected to 1) hydration for 1 h, 24 h, or 48 h for the control batch, batch B, or batch D respectively; and 2) hydration for 1 h, 24 h, or 48 h with freeze thawing for 5-cycles for batch A, batch C, or batch E, respectively. These formulations were stored at 10 +/- 2 degrees C and 37 +/- 0.1 degrees C. Small unilamellar vesicles (SUV) and MLV were prepared using L-alpha phosphatidylcholine (PC), and buffer at pH 5.0, 5.5, 6.0, 6.5, and 7.0, containing 80 mg hydroxyzine or 82 mg cetirizine by the ethanol injection and thin-lipid film hydration methods, respectively. These formulations were stored at 10 +/- 2 degrees C. Liposomes were evaluated immediately after preparation and after storage by determining percent entrapment of hydroxyzine (PETH) or of cetirizine (PEC) and by observing changes in the physical appearance (PA). Particle size (PSA) of the liposomes freshly prepared at pH=6.5 was measured from transmission electron micrographs (TEM). RESULTS: Increasing thin-film hydration time or repeated freeze-thawing did not affect the initial PETH or long-term stability of control, A, B, C, D, and E batches of MLV containing hydroxyzine stored at 10 +/- 2 degrees C. At 37 +/- 0.1 degrees C, PETH of all MLV batches decreased considerably after 1 month. This was more evident in batches B, C, and E exposed to freeze-thawing. The PETH of SUV increased markedly from 53.0% to 84.0% when the pH of the buffer was increased from 5.0 to 5.5. As pH increased from 6.0 to 7.0, PETH continued to increase from 84% to 94%. The initial PETH of MLV increased slightly from 82.0% to 94.0% as the buffer pH values increased from 5.0 to 7.0. There was no effect of pH on initial PEC, and stability of SUV or initial PEC of MLV, which ranged from 92% to 94%, as buffer pH values increased from 5.0 to 6.5. After storage at 10 +/- 2 degrees C PEC in MLV decreased from 94% to 74%. CONCLUSIONS: The freeze-thawing processes had some effect on the stability of liposomes stored at temperatures higher than ambient temperature, 37 +/- 0.1 degrees C. The effect of changing the buffer pH from 5.5 to 7.0, and from 5.0 to 6.5 on initial PETH and PEC, respectively, was minimal. After 24 months at l0 +/- 2 degrees C, pH had no effects on PETH; however, PEC of MLV decreased. PMID- 15830725 TI - Oral bioadhesive drug delivery systems. AB - The oral mucosal cavity is a feasible, safe, and very attractive site for drug delivery with good acceptance by users. The mucosa is relatively permeable and robust, shows short recovery times after stress or damage, is tolerant to potential allergens, and has a rich blood supply. Moreover, oral mucosal drug delivery bypasses the first-pass effect and avoids presystemic elimination in the gastrointestinal tract. Bioadhesive systems provide intimate contact between a dosage form and the absorbing tissue, which may result in high concentration in a local area and hence high drug flux through the absorbing tissue. The efficacy of oral bioadhesive drug delivery systems is affected by the biological environment and the properties of the polymer and the drug. In the present paper, we review systematically some relevant citations regarding the environment, strategies for oral drug delivery and evaluation, and utilization of the main polymers. PMID- 15830726 TI - Cross-linked cationic polymer microparticles: effect of N-trimethyl chitosan chloride on the release and permeation of ibuprofen. AB - Microparticles made by cross-linking hydrophilic polymers, such as chitosan, have been used to modify the release rate of a loaded drug. In this study a polymer with fixed positive charges, N-trimethyl chitosan chloride (TMC), was used in combination with chitosan to formulate microparticles to investigate its effects on drug release rate and transport across intestinal epithelial cells. The microparticles were prepared by cross-linking these cationic polymer(s) using sodium citrate as the ionic cross-linker. This process was done under homogenization and ultrasonication to control the size of the particles. The addition of TMC to the chitosan microparticles resulted in an increase in particle size of the microparticles and an increase in ibuprofen release rate as compared to the microparticles containing chitosan alone. Permeation of ibuprofen across Caco-2 cell monolayers, after administration of a suspension of the microparticles to the apical side, was not significantly different for the microparticles containing TMC as compared to those consisting of chitosan alone. It was concluded that release of TMC molecules from the microparticles was probably not sufficient to interact with the intestinal epithelial cells in order to change the permeation of the released drug. PMID- 15830727 TI - Nanosuspension formulations for low-soluble drugs: pharmacokinetic evaluation using spironolactone as model compound. AB - Various particle sizes of spironolactone as a model low solubility drug were formulated to yield micro-and nanosuspensions of the type solid lipid nanoparticles and DissoCubes. Seven oral and one i.v. formulations were tested in an in vivo pharmacokinetic study in rats with the aim of characterizing the bioavailability of spironolactone on the basis of its metabolites canrenone and 7 alpha-thiomethylspirolactone. In addition, a dose escalation study was carried out using nonmicronized spironolactone suspension as well as a nanosuspension type DissoCubes. On the basis of AUC as well as Cmax ratios, three groups of formulations were distinguished. The biggest improvement was seen with a solid lipid nanoparticle formulation yielding a 5.7-fold increase in AUC for canrenone and a similar improvement based on the Cmax metric, followed by a group of three formulations containing nanosized, micronized, and coarse drug material and surfactant. The DissoCubes nanosuspension yielded highly significant improvements in bioavailability averaging 3.3-fold in AUC and 3.0-fold in terms of Cmax for canrenone. The third class encompasses all other formulations, which showed very little to no improvement in bioavailability. The results show that the particle size minimization was not the major determining factor in the bioavailability improvement. Rather, the type of surfactant used as stabilizer in the formulations was of greater importance. Improvement in drug solubility in the intestine as well as in dissolution rate of spironolactone are the most likely mechanisms responsible for the observed effect, although additional mechanisms such as permeability enhancement may also be involved. PMID- 15830728 TI - Assessing contemporary substance abusers with the MMPI MAC Andrews Alcoholism Scale: a review. AB - We reviewed 71 United States-based MacAndrew Alcoholism Scale (MAC), as revised (MAC-R) studies totaling almost 32,000 Ss, with adolescent and adult substance abusers, from studies published since the last MAC reviews (1989) through 2001. Results suggest that the MAC, and to some extent, the MAC-R, significantly correlates with measures of alcohol and substance abuse in both male and female adolescents and adults, across a diverse spectrum of the use-abuse continuum. Nonclinical groups (100%) scored below the clinical ranges on the MAC/MAC-R, while 79% of adolescent substance abusing groups scored > R 23, indicative of problems with substance abuse. Persons who abused alcohol, drugs, and polydrugs had mean MAC/MAC-R scores > 23, which ranged from 77% to 100% of the cases. The MAC/MAC-R does well in discriminating persons who abuse substances compared to nonclinical, nonabusing groups, but appears to lose diagnostic efficiency with psychiatric patients, and especially with medical patients with seizure disorders. Using R > 25 seems to improve diagnostic accuracy with this population. Meaning of false positives and false negatives were explored and discussed. PMID- 15830729 TI - Binarisms, regressive outcomes and biases in the drug policy interventions: a theoretical approach. AB - The golden age of drug policy was characterized by the informal regulation of drug use. Formalization of the control over regulation and its increasingly strict, aggressive character led to the emergence of a binary attitude. The main binarisms: pharmaceutical or drug; ban or tolerance; punishment or treatment; psychopathological or pathopsychological approach; subjective or objective knowledge; traditional or alternative. On the basis of Kuhn's paradigm theory, these binarisms can be integrated. Drug policy interventions based on the binary attitude have had regressive effects. Using the work of Sam Sieber, the author distinguishes nine regressive influences: functional imbalance, perverse diagnosis, ricochet, overload, goal displacement, exploitation, provocation, classification, and placation. The regressive influences have caused the escalation of "the drug problem," which in turn has led to further regressive interventions. This vicious circle could be broken by eliminating the four biases -the paternalistic, elitist, rationalist, and activist biases--underlying the regressive interventions. PMID- 15830730 TI - Trihexyphenidyl (Artane): a Brazilian study of its abuse. AB - In Brazil, the medicinal misuse of trihexyphenidyl (Artane) has been observed among several segments of society. The present study was conducted in the city of Sao Paulo during 2002 to characterize this abuse. A sample of 21 users and 16 ex users was interviewed using qualitative methodology; the subjects were single, unemployed, male polydrug users, who used trihexyphenidyl in order to attain states of mental alterations, mainly hallucinations and deliriums. Trihexyphenidyl is consumed in association with alcohol, other licit drugs (benzodiazepines), or illicit drugs, impairing cognitive functions such as memory, attention, and learning, intervening with some activities of users' daily life. PMID- 15830731 TI - A modified therapeutic community for homeless persons with co-occurring disorders of substance abuse and mental illness in a shelter: an outcome study. AB - This article reports on a study conducted to determine the effectiveness of a modified therapeutic community (MTC) shelter on client outcomes. The seven-study hypotheses focused on whether greater effectiveness in the MTC would be demonstrated in longer periods of sobriety, fewer days of psychiatric hospitalization, shorter lengths of stay in a shelter, positive discharge from the shelter, medication compliance, housing placement within the first year, and appropriate housing placement according to level of functioning. The study utilized a quasi-experimental design with two groups: 1) an experimental group (E) homeless persons with co-occurring disorders (COD) of substance abuse/dependence and mental illness who reside in a modified therapeutic community (N = 70); and 2) a comparison group (C) of veterans with CODs living in a general shelter (N = 70). The data collection procedures involved a retrospective review of closed case records for subjects in the facilities from September 1, 1998-June 1, 2000 for the MTC shelter, and from June 1, 1999-June 1, 2000 for the general shelter. Baseline differences between the E and C groups were found in age, length of homelessness, years of education, years of work experience, veteran status, marital status, and psychiatric diagnosis; all except for veteran status were unrelated to outcomes and were controlled in subsequent analyses. Significant difference was found on medication compliance when controlled for both groups. Overall, this study showed some promise for the MTC approach. The study also raised a question as to the contribution of veteran status to the differences between groups and to treatment of CODs. PMID- 15830732 TI - Predictors of benzodiazepine discontinuation in subjects manifesting complicated dependence. AB - We described characteristics of subjects with benzodiazepine dependence that was typically complicated by harmful and hazardous alcohol use or high benzodiazepine doses, and assessed predictors of successful discontinuation of benzodiazepines for this group. Seventy-six patients who participated in a randomized clinical trial of two different gradual withdrawal treatment approaches were assessed. The trial was conducted between February 1995 and July 1999. The mean age +/- SD of subjects was 40.0 +/- 9.6 years, 55% were male, 38% were married or cohabiting, and 70% had received more than nine years of education. The median benzodiazepine dose was 35 mg/day (range 2.5-180) in diazepam equivalents. The median duration of benzodiazepine use was 84 (range 8-360) months. Subjects with lower benzodiazepine doses and no previous withdrawal attempts were more successful at benzodiazepine discontinuation. Cluster B personality/borderline personality disorder was associated with an inability to stop benzodiazepine use and with "dropping out" of treatment. Alcohol use-related disorders or other psychiatric diagnoses were not associated with outcome. Further studies on predictors of successful benzodiazepine discontinuation in different populations are required. Patients manifesting cluster B personality/borderline personality disorder and benzodiazepine dependence may need concomitant treatment for their personality disorders to benefit from benzodiazepine discontinuation treatment. PMID- 15830733 TI - Gender effects following repeated administration of cocaine and alcohol in humans. AB - RATIONALE: Use of cocaine, alcohol, and the two drugs simultaneously is common and the risk of morbidity and mortality associated with these drugs is widely reported. This double-blind, placebo-controlled, randomized study examined gender differences in response to administration of these drugs alone and in combination. METHODS: Current users of cocaine and alcohol (n = 17) who met diagnostic criteria (DSM-IV) for cocaine dependence and alcohol abuse or dependence (not physiologically dependent on alcohol) and who were not seeking treatment for substance use disorders gave voluntary, written, informed consent to participate in three drug administration sessions:1) four doses of intranasal cocaine (1 mg/kg every 30 min) with oral alcohol (1 g/kg following the initial cocaine dose and a second drink at +60 min (120 mg/kg) calculated to maintain a plasma alcohol concentration of approximately 100 mg/dL; 2)four doses of cocaine and alcohol placebo; 3) cocaine placebo and alcohol. Pharmacokinetics were obtained by serial blood sampling, physiological measurements (heart rate and blood pressure) were obtained with automated equipment, and subjective effects were assessed using visual analog scales over 480 min. RESULTS: Responses to cocaine, alcohol, and cocaine-alcohol were equivalent by gender for most measurements. Women had higher heart rates following alcohol administration (p = .02). Women consistently reported higher ratings for "Feel Good:' a measure of overall mental/physical well-being, for all study conditions, reaching statistical significance for cocaine (p = .05) and approaching significance for alcohol administration (p = .1). CONCLUSION: Women showed equivalent responses to drug administration with the exception of perception of well-being, which was significantly increased for women. These findings may have implications for differential risk for acute and chronic toxicity in women. PMID- 15830734 TI - A bio-behavioral model of addiction treatment: applying dual representation theory to craving management and relapse prevention. AB - A bio-behavioral approach to drug addiction treatment is outlined. The presented treatment model uses dual representation theory as a guiding framework for understanding the bio-behavioral processes activated during the application of expressive therapeutic methods. Specifically, the treatment model explains how visual processing techniques can supplement traditional relapse prevention therapy protocols, to help clients better manage cravings and control triggers in hard-to-treat populations such as chronic substance-dependent persons. PMID- 15830735 TI - The development of depression during psychodynamic group psychotherapy among subjects in substance abuse remission. AB - The aim of the present study was to examine the development of self-reported depression during psychodynamic group therapy among patients manifesting a history of substance use-related problems between 1993-2000. Subjects (n = 100) were prospectively followed with respects to depression levels at intake and after 6, 12, and 18 month in therapy. Depression was measured with the Beck Depression Inventory. Overall, there was a significant decrease in depression after 18 months in therapy. The development of depression in different subgroups was also analyzed. PMID- 15830736 TI - A comparison of the Yale-Brown Obsessive Compulsive Scale for "heavy drinking" with a single item craving measure: construct validity and clinical utility. AB - The measurement of alcohol "craving" began with single-item scales. Multifactorial scales developed with the intention to capture more fully the phenomenon of craving. This study examines the construct validity of a multifactorial scale, the Yale-Brown Obsessive Compulsive Scale for heavy drinking (Y-BOCS-hd). The study compares its clinical utility with a single item visual-analogue craving scale. The study includes 212 alcohol dependent subjects (127 males, 75 females) undertaking an outpatient treatment program between 1999 2001. Subjects completed the Y-BOCS-hd and a single item visual-analogue scale, in addition to alcohol consumption and dependence severity measures. The Y-BOCS hd had strong construct validity. Both the visual-analogue alcohol craving scale and Y-BOCS-hd were weakly associated with pretreatment dependence severity. There was a significant association between pretreatment alcohol consumption and the visual-analogue craving scale. Neither craving measure was able to predict total program abstinence or days abstinent. The relationship between obsessive compulsive behavior in alcohol dependence and craving remains unclear. PMID- 15830737 TI - Beliefs and social norms about cigarettes or marijuana sticks laced with embalming fluid and phencyclidine (PCP): why youth use "fry". AB - Recent drug-use monitoring among Houston adolescents has detected a concoction of cigarettes or marijuana sticks laced with embalming fluid and PCP ("fry"). To shed light on this mixture, the current pilot study used a qualitative approach to investigate relevant beliefs and norms associated with fry initiation and perceived addiction among 38 youth who were attending outpatient and inpatient drug-user treatment programs in the spring of 2003. Respondents perceived that addiction to fry could occur as early as initial consumption, and the majority of participants indicated that their second fry event occurred either the same day as their initial use or the next day. In addition, fry use was perceived to have extremely dangerous consequences. Youth stated that users have impaired motor skills, hallucinations, long-term mental health problems, incoherent behavior, paranoia, and aggressive behaviors. Implications for these results are discussed. PMID- 15830738 TI - Patterns of alcohol consumption in a Northern Irish sample. AB - This paper examines the drinking habits of a Northern Irish sample during a six month period in 1998. In addition the study examines the influence of contextual variables on the quantity and frequency of alcohol consumption. Questionnaires were administered to 600 participants; the response rate was 39.8% (239). An unexpected low abstinence rate was observed that, however, may be due to response bias. The results revealed high frequency (29.7% drink on four or more days a week) and high quantity of alcohol consumption (mean units per week 43.21, SD 40.33). Beer drinkers consumed the largest quantity of alcohol and also had the highest frequency of alcohol consumption. It was observed that 45.8% of all drinking events took place in a public bar and the popularity of the public bar for alcohol consumption was not influenced by age or gender. The present investigation revealed that almost half (45%) of individuals consume more than one type of beverage at one sitting, and there is a trend of consuming alcohol in more than one place during a single drinking session. These results indicate a distinctive drinking pattern within Northern Ireland and have implications for studies investigating the effects of alcohol on the social drinker. PMID- 15830739 TI - Health services that do not serve: promoting, rather than preventing, tuberculosis. PMID- 15830740 TI - Tuberculosis bacteriology--priorities and indications in high prevalence countries: position of the technical staff of the Tuberculosis Division of the International Union Against Tuberculosis and Lung Disease. AB - Smear microscopy for acid-fast bacilli (AFB) remains the first priority for national tuberculosis programmes (NTPs) in high-prevalence countries. No other established technique offers the same advantages of accuracy, speed, appropriateness and accessibility. Its sensitivity may be reduced in HIV-positive cases or because of technical deficiencies, and it lacks specificity for viable bacilli in follow-up examinations. Its main problem is that it is tedious, necessitating an effective external quality assessment (EQA) system following international guidelines. Operational research is needed to optimise the staining he place of sputum concentration o define t and fluorescence microscopy, to challenge difficult and obsolete strategies and to streamline procedures. Culture is much more difficult to set up and is usually impossible to decentralise. Because of its lower yield and higher costs, its efficiency for case detection in NTPs will lag well behind that in industrialised countries. The technique should only be used as a preliminary to drug susceptibility testing (DST). DST should not be developed to the detriment of the AFB microscopy network and its EQA. It should be used mainly for monitoring drug resistance. Continuous monitoring of resistance in a representative sample of isolates from first-line failure and relapse cases may be more efficient and more accurate than periodic surveys among new cases, and can be used to identify MDR-TB, whose treatment should be standardised, because of considerable risk of error in the laboratory. A specialist service offering molecular techniques may be useful for exceptional cases, but it has no place in the routine work of NTPs. PMID- 15830741 TI - Nutrition and lung health. AB - Several lung diseases have been associated with oxidative stress and linked to oxidant insults such as cigarette smoke, air pollutants and infections. Consequently, dietary factors and nutrients with a potential protective role in the oxidative process and inflammatory response have been implicated in the genesis or evolution of these diseases. These nutrients include fruits and vegetables, antioxidant vitamins such as vitamin C, vitamin E, betacarotene and other carotenoids, vitamin A, fatty acids and some minerals such as sodium, magnesium and selenium. This article reviews the potential mechanisms by which dietary factors may affect respiratory health, and discusses epidemiological evidence for the link between diet and lung diseases. Most of the available evidence on the effect of dietary factors on the risk for obstructive lung diseases are derived from cross-sectional studies. These studies suggest that antioxidant vitamins, particularly vitamin C, and to a lesser extent other antioxidants, have a protective effect against lung diseases. However, the few intervention studies have not been conclusive. High intake of fresh fruit and some vegetables appears to have a beneficial effect on lung health and their consumption should be recommended on a daily basis. Supplementation of vitamin C and other antioxidants could be proposed in subjects with additional oxidative stress challenge, such as exposure to high levels of air pollution. Subjects with impaired immune response could also benefit from vitamin A and zinc supplementation. Further studies are needed to determine the impact of diet on the incidence and evolution of lung diseases. PMID- 15830742 TI - Role of mycobacteria-induced monocyte/macrophage apoptosis in the pathogenesis of human tuberculosis. AB - Apoptosis is a physiological programmed cell death process whose dysregulation plays an important role in different human infectious diseases. An increasing number of intracellular pathogens are known to induce target cell apoptosis, while some other parasites inhibit it. Unlike necrosis, apoptosis is a silent immunological event occurring without inflammation. Infection-induced target cell apoptosis may be a successful strategy to eliminate pathogens and assure host survival. Conversely, apoptosis inhibition could represent an adaptive mechanism for pathogen survival, while it may be beneficial for the host to initiate an effective immune response. The worldwide increase in tuberculosis has stimulated more research aimed at defining the interaction between Mycobacterium tuberculosis and the immune system. M. tuberculosis possesses sophisticated strategies to circumvent its fate within target monocytic cells. Apoptosis of alveolar macrophages and monocytes has been described as a consequence of M. tuberculosis infection. Moreover, the observation that mycobacterial lipoproteins activate macrophages through Toll-like receptor (TLR) 2 suggests that innate immune receptors contribute to defence against M. tuberculosis. There is evidence that TLR-induced apoptosis modulates inflammation and immune activation during M. tuberculosis infection. Finally, the role of apoptotic-infected cells as a source of microbial antigens for cross-priming of effector T-cells is also discussed. PMID- 15830743 TI - A method to determine the utility of the third diagnostic and the second follow up sputum smear examinations to diagnose tuberculosis cases and failures. AB - SETTING: Forty-two laboratories in four countries. OBJECTIVE: To determine the number of sputum smear examinations required to identify one additional case of tuberculosis from a third serial diagnostic smear or one additional treatment failure from a second serial follow-up smear. MATERIAL AND METHODS: Country specific prevalence of new cases and failures among 59 665 examinees were determined, as well as the incremental yield from serial smears. The reciprocal value of the product of the prevalence of cases or failures and the respective incremental yield from the last serial smear provided the number of slides that have to be examined to identify one additional case or failure. RESULTS: The expected prevalence of cases among suspects ranged from 5.4% to 32.8%; the incremental yield from a third serial smear ranged from 0.7% to 7.2%. Between 122.7 and 796.3 smears were required to identify one additional case with the third serial smear. The prevalence of failures among follow-up examinees ranged from 1.0% to 2.5%; the incremental yield from the second follow-up serial smear ranged from 4.5% to 26.9%. Between 164.8 and 2133.4 slides were required to identify one additional failure with the second serial smear. CONCLUSION: The utility of serial smears can be rationally determined by careful review of program data. PMID- 15830744 TI - Tuberculosis at Chris Hani Baragwanath Hospital: numbers of patients diagnosed and outcomes of referrals to district clinics. AB - SETTING: Chris Hani Baragwanath Hospital, in Johannesburg. South Africa is experiencing a serious epidemic of tuberculosis (TB), with a measured rate of 500 cases per 100,000 population. Patients in public hospitals are referred for treatment to district clinics or to specific TB hospitals. OBJECTIVES: To measure numbers of patients diagnosed with TB, and to describe the type of disease and referral outcomes. METHODS: A descriptive study of patients diagnosed with TB in a tertiary hospital. Data in patient files were collected for medical and paediatric patients, and patients were interviewed by researchers who visited clinics to check if referred patients had attended. RESULTS: In the 8-week study period, 1291 patients were diagnosed with TB: 74% had pulmonary disease and 80% of those tested for HIV were positive; 19% died in hospital, and 21% required transfer to a TB hospital. Only half of the patients referred to clinics attended within 2 weeks. CONCLUSIONS: Patients with advanced TB are commonly seen at this hospital. Notifications were an inadequate record and an unacceptable proportion of patients were lost between hospital and clinics. This study has recommended that opportunities for education and discussion must be provided if patients are to reach clinics successfully. PMID- 15830745 TI - Planning for PPM-DOTS implementation in urban slums in Kenya: knowledge, attitude and practices of private health care providers in Kibera slum, Nairobi. AB - SETTING: Kibera, the largest slum in Nairobi, Kenya. OBJECTIVE: To determine the tuberculosis (TB) knowledge, attitude and practices (KAP) of private health care providers (PHCPs) to identify their training needs and willingness to participate in a National Leprosy and Tuberculosis Control Programme (NLTP) guided TB control effort in the slum. DESIGN AND METHODOLOGY: A cross-sectional survey. The KAP of PHCPs was assessed using an interviewer administered questionnaire. RESULTS: Of 75 PHCPs interviewed, the majority (96.0%) were paramedics; 51 (77.1%) did not consider sputum smear microscopy crucial in patients presenting with prolonged cough or when a chest X-ray was suggestive of TB; of 29 (38.7%) who indicated familiarity with the drugs used in TB treatment, 20 (58.5%) would have chosen the NLTP-recommended regimens for the treatment of the various types of TB; 16 (21.3%) PHCPs indicated that they treated TB, six (37.5%) of whom were not familiar with anti-tuberculosis drug regimens. All the PHCPs referred TB suspects to the public sector for diagnosis. CONCLUSION: This study reveals a significant gap in TB knowledge among the PHCPs in Kibera slum. However, given appropriate training and supervision, there is potential for public-private mix for DOTS implementation in this setting. PMID- 15830746 TI - Determining TB rates and TB case burden for refugees. AB - SETTING: DeKalb County, Georgia. OBJECTIVES: To calculate and compare tuberculosis (TB) rates in refugees to US-born, total foreign-born (refugee and other), and other foreign-born persons and to determine the contribution of refugees to the county TB case burden. METHODS: The study included: (1) collection of county TB case numbers and population figures from 1995 through 1999; (2) estimation of the refugee population; (3) comparison of TB rates; and (4) calculation of the refugee TB case burden. Sensitivity analysis was performed on refugee population estimates. RESULTS: From 1995 through 1999, estimating that refugees made up 10% of the foreign-born population, the average TB rate for refugees was 83.2 per 100,000, compared with 12.7 for US-born persons. From 1997 through 1999, refugees had a seven-fold greater risk of having TB than US-born persons and a two-fold greater risk than other foreign-born persons. Refugees represented respectively 7.6% and 19.3% of the county and foreign-born TB case burdens. For TB rates to be equal among all foreign-born persons, refugees would need to make up 15-25% of the foreign-born population. CONCLUSION: Despite overseas screening, refugees have high TB rates, and contribute substantially to the county TB case burden. Enhanced surveillance and targeted programs to address TB in refugees should be a public health priority. PMID- 15830747 TI - Structured behavioral intervention to increase children's adherence to treatment for latent tuberculosis infection. AB - SETTING: San Diego, California public health department tuberculosis clinic. OBJECTIVE: To evaluate the effectiveness of a behavioral intervention (the Treasure Chest) that employed the behavior modification techniques of self monitoring and positive reinforcement to increase adherence to therapy for latent tuberculosis infection (LTBI) in children. DESIGN: A retrospective study utilizing a historical comparison group. The treatment adherence of children who attended San Diego County TB Control Program (SDCTBCP) clinics with positive tuberculin skin tests who were started on LTBI treatment prior to Treasure Chest program implementation (n=841) was compared to that of SDCTBCP children enrolled in the Treasure Chest program (n=741). RESULTS: Logistic regression analysis demonstrated that children who participated in the Treasure Chest program were 2.4 times more likely to complete therapy than those who did not receive the intervention. CONCLUSION: The Treasure Chest intervention is effective in increasing adherence to LTBI therapy in children. PMID- 15830748 TI - Retreatment management strategies when first-line tuberculosis therapy fails. AB - SETTING: Public ambulatory centers in northern Lima, Peru. OBJECTIVE: To compare two retreatment strategies in Category I failures. DESIGN: Retrospective cohort study of Category I failures enrolled between February 1997 and October 2001. Strategy A was a nationwide approach, applying a Category II regimen; if that regimen failed, a standardized regimen including second-line drugs was used. Strategy B was a pilot protocol designed to diagnose and treat multidrug resistant tuberculosis (MDR-TB); this strategy included drug susceptibility testing (DST) and eliminated the Category II regimen. RESULTS: Of 125 patients that Category I failed to cure, 73 entered Strategy A and 52 entered Strategy B. Almost 90% of those with DST results had MDR-TB. Strategy B was three times more likely than Strategy A to cure patients (79% vs. 38%, RR = 2.9, 95% CI 1.7-5.1) and five times more likely to cure patients than the Category II regimen alone (79% vs. 15%, RR 5.2, 95% CI 3.0-9.2). Strategy B also significantly reduced delays to MDR-TB diagnosis and to the initiation of MDR-TB therapy. CONCLUSIONS: Under program conditions, a retreatment strategy based on DST and eliminating the Category II regimen can improve clinical outcomes among Category I treatment failures found to have active, infectious MDR-TB. PMID- 15830749 TI - Clinical and epidemiological aspects of smoking and tuberculosis: a study of 13,038 cases. AB - SETTING AND OBJECTIVE: Smoking habit and tuberculosis (TB) appear to have common epidemiological and clinical links. The present study was to evaluate risk factors in TB patients with and without a smoking habit. DESIGN: A cross sectional observational study of cases (TB with smoking) and controls (TB without smoking) from the same registry. Data were retrieved from case notes and interviews of patients registered in the Tuberculosis Control Programme in Cataluuna, Spain, between 1 January 1996 and 31 December 2002. Statistical analyses included univariate and stepwise multiple logistic regression analyses. RESULTS: There were 13,038 recorded patients. Social factors associated with TB in smokers were male sex (adjusted odds ratio [aOR] 2.2), age <64 years, alcohol use >30 g/day (aOR 7.4) and intravenous drug use (aOR 1.5). Smokers developed more pulmonary disease (aOR 1.5) and more cavitary lesions (aOR 1.9), and were more likely to require hospitalisation (aOR 1.8) which was more protracted. Differences in mortality and delay in diagnosis did not reach statistical significance. CONCLUSIONS: Smoking leads to faster and more severe progression of TB. The cost of TB-related hospitalisation for smokers increases by approximately one million euros per year. PMID- 15830750 TI - Tuberculosis mortality by industry in the United States, 1990-1999. AB - OBJECTIVE: To identify occupations and industries with elevated respiratory tuberculosis (TB) mortality in the United States for the period 1990-1999, we used National Center for Health Statistics multiple-cause-of-death data, restricted to certain states for which information on decedents' usual industry and occupational information was available and limited to US residents aged > or =15 years. DESIGN: A total of 7686 deaths between 1990 and 1999 were attributed to respiratory TB. Proportionate mortality ratios (PMRs), adjusted for age, sex, and race, were calculated from US census occupation and industry classifications. RESULTS: Industries and occupations involving potential contact with infected cases (e.g., health care workers), those with silica exposure and silicosis (e.g., mining and construction), and those associated with low socioeconomic status had significantly elevated TB mortality. CONCLUSIONS: Overall, the pattern of findings echoes that described in various prior reports, which indicates that the potential for exposure and disease development still persists among certain worker groups. The findings should be useful in guiding occupationally targeted TB prevention programs. PMID- 15830751 TI - Directly observed treatment of tuberculosis in Hong Kong. AB - OBJECTIVE: To evaluate the local use of out-patient directly observed treatment (DOT) for tuberculosis (TB) control. SETTING: Seventeen government chest clinics managing around 80% of TB patients in Hong Kong. METHOD: A retrospective cohort. TB patients registered for treatment from 1 July to 31 August 2000 were followed up for 2 years. Baseline socio-demographic and clinical characteristics were correlated with adherence to DOT, mode of treatment and treatment outcomes. RESULTS: Of 988 patients, respectively 142, 140 and 21 switched to non-DOT within 2 months, 2-6 months and after 6 months. More Chinese patients than ethnic minorities switched to non-DOT within the first 2 months (15.2% vs. 0%, P = 0.001). Geographical inconvenience (31.7%) was the primary reason for switching initially, with increasing proportions giving no specific reason after the second month. Patients staying on DOT in the first 2 months had a significantly higher cure rate than those not on DOT, in both univariate (92.7% vs. 83.9%, P = 0.002) and multivariate analysis (OR = 2.5, P = 0.001). Subsequent switching, intermittent or daily regimen did not appear to affect the outcome. CONCLUSION: Significant numbers of patients failed to stay on DOT, and those staying initially had a better outcome. PMID- 15830752 TI - Analysis of factors affecting the epidemiology of tuberculosis in China. AB - SETTING: The tuberculosis (TB) epidemic situation is both a public health problem and a socio-economic issue in China. OBJECTIVE: To examine the effects of socio economic development and of the TB control strategy on the TB epidemic in China. METHODS: Based on the four National Epidemiological Surveys of TB and the indices of socio-economic development in China, correlation co-efficiency was used to analyse the relationship between changes in the TB epidemic situation, the socio economic level and the Health V TB control Project. RESULTS: The prevalence of smear-positive TB had significant medium correlation with the per capita net income of the rural population, the consumption level of the urban population, the per capita GDP, the population density, and the proportion of rural to total population, among which the correlation with the first four was negative and with the last was positive. The decline in prevalence in the project areas was much greater than in the non-project areas (44.4% vs. 12.3%), while their GDP increases were similar. CONCLUSION: With socio-economic development, correlation between the socio-economic indices and the TB epidemic becomes more significant. The TB control project is vital to reduce the prevalence of TB in China. PMID- 15830753 TI - Diagnostic usefulness of bone marrow aspiration material for the amplification of IS6110 insertion element in extrapulmonary tuberculosis: comparison of two PCR techniques. AB - SETTING: In many cases of extra-pulmonary tuberculosis (EPTB), with the exception of paucibacillary analysed specimens, the suspected site of mycobacterial infection is relatively inaccessible or unknown, making laboratory confirmation of TB laborious and problematic. OBJECTIVE: Two different polymerase chain reaction (PCR) based methods were compared to investigate the validity of bone marrow aspiration material as an easily accessible alternative sample for molecular analysis in EPTB. DESIGN: We amplified the same sequence of IS6110 of Mycobacterium tuberculosis complex in 19 confirmed cases of EPTB using two different nested PCR techniques: one in-house 'classic' PCR and another based on LightCycler technology. RESULTS: Both methods demonstrated the same reliability when performed in samples of infected tissue. However, the LightCycler protocol was superior to the in-house system when applied in bone marrow aspiration material, revealing positivity in 18/19 compared to 13/19 samples of 'classic' PCR. CONCLUSION: The application of an optimised LightCycler nested amplification protocol in bone marrow aspirates may promote diagnostic accuracy in difficult and/or urgent cases of EPTB. PMID- 15830755 TI - A survey of tuberculosis hospitals in India. PMID- 15830754 TI - IgA antibody responses to Mycobacterium tuberculosis recombinant MPT-64 and MT 10.3 (Rv3019c) antigens in pleural fluid of patients with tuberculous pleurisy. AB - SETTING: Microbiological tests lack sensitivity for pleural tuberculosis (TB) and histopathology is expensive, time consuming and needs specialised personnel. Immunoassay (ELISA) may be a promising approach in this respect. OBJECTIVE: To evaluate the reactivity of IgA antibody to MPT-64 and MT-10.3 recombinant mycobacterial protein antigens in pleural fluid as a marker of pleural TB, based on the fact that IgA is the main antibody in the mucosa/serosa of the gastrointestinal and respiratory tract. METHOD: Anti-MPT-64 and MT-10.3 IgA response was determined by ELISA in 72 patients with pleural TB and 27 with other pleural conditions. RESULTS: High sensitivities for IgA were measured against MPT 64 (52/72, 72%) and MT-10.3 (52/72,72%) antigens. Combining the sensitivities of both antigens, further increase in sensitivity (55/72, 76%) was obtained with no loss of specificity (96%). Similar IgA reactivity was obtained from culture negative and culture-positive specimens. In eight pleural TB patients with human immunodeficiency virus (HIV) co-infection, the sensitivity was 88% (7/8). CONCLUSION: To our knowledge, this is the first description of the presence of IgA antibody pleural TB effusion reactive to MPT-64 and MT-10.3, with sensitivity similar to histopathological examination, which is presently considered the gold standard for pleural TB. PMID- 15830756 TI - Comprehensive cost description of tuberculosis care. PMID- 15830757 TI - Partial interferon-gamma receptor deficiency and disseminated tuberculosis. PMID- 15830759 TI - The "cannot-intubate, cannot-ventilate" scenario. PMID- 15830758 TI - Bovine tuberculosis in Africa. PMID- 15830760 TI - Unknown contributions of Arab and Islamic scholars in the field of anesthesia in the West. PMID- 15830761 TI - Perioperative beta-blocker therapy: option or obligation? PMID- 15830762 TI - Initial management of traumatic brain injury. PMID- 15830763 TI - Intensive care management of traumatic brain injury. PMID- 15830764 TI - Diastolic dysfunction complicating anesthetic management. AB - Coronary artery disease combined with constrictive pericarditis results in diastolic dysfunction with a well-preserved systolic function. A case of constrictive pericarditis with a restrictive filling pattern and a well established preoperative diastolic dysfunction, that had an elective pericardiectomy and an off pump coronary artery by pass graft, is discussed. Once revascularization was performed, introduction of Inj. nitroglycerine resulted in a dramatic improvement in the hemodynamics, along with a drop in the intra chamber filling pressures, indicating an improvement in the diastolic function. The role of revascularization and vasodilators as therapeutic tools in the management of diastolic dysfunction in such cases, is highlighted. PMID- 15830765 TI - Morbid obesity--a review. AB - Obesity, the most common nutritional disorder in the United States, has taken on epidemic proportions, spreading rapidly during the 1990s across all states, regions, and demographic groups in the United States, according to the CDC. The presence of coexisting disease, difficulty with airway management and respiratory compromise, as well as alterations in drug metabolism in obese patients, combine to make understanding this particular disorder vital for the clinical anesthesiologist. This review summarizes the clinical presentation, pathophysiology, and treatment of morbid obesity. Preoperative assessment, intraoperative management, and postoperative care, as well as anesthetic management of coexisting disease states of obesity are addressed. PMID- 15830766 TI - The cocaine-using patient: perioperative concerns. AB - Cocaine is an extremely addictive agent which can produce stimulation of the sympathetic nervous system due to the inhibition of catecholamine reuptake at the synaptic junction. Probability of a patient presenting to the operating theatre with acute cocaine intoxication is increasingly likely. The physiological effects of even chronic cocaine abuse on various organ systems have an impact on anesthetic management. A preoperative review of major organ systems is essential. Nitroglycerin has been used in the management of hypertension associated with coronary vasoconstriction. Controversy exists regarding management of ventricular dysrhythmias and asystole. The use of epinephrine to treat asystole is controversial in the presence of a state of excess catecholamines induced by cocaine. General anesthesia may include barbiturates, nitrous oxide, and opioids, but inhalational agents must be used with caution due to their myocardial depressant effects. Regional anesthesia may be a good choice if coagulopathies and hypovolemia are corrected. PMID- 15830767 TI - Regional analgesia combined with avoidance of narcotics may reduce the incidence of postoperative vomiting in children. AB - BACKGROUND: The anesthesia literature cites a high incidence of postoperative vomiting (POV) after pediatric ochidopexy and hernia repair (34-50%) and after penile procedures (37-49%). We hypothesized that regional analgesia combined with avoidance of narcotics administered to children scheduled for lower abdominal or urologic procedures may be associated with a lower incidence of POV. The aim of this prospective study was to 1) assess the incidence of POV in children in the hospital and during a 24-h post-anesthesia study period, and 2) evaluate the effect of age on POV. METHODS: After obtaining institutional and parental consent, 110 pediatric outpatients, 1-12 yr old, ASA physical status I or II, scheduled for elective outpatient urologic or lower abdominal procedures, were enrolled. Children were fasting and premedicated with midazolam, 0.5 mg/kg p.o. They received a combined light general anesthesia and a presurgical caudal block. Anesthesia was induced via a mask and consisted of halothane and nitrous oxide in oxygen. For the caudal block 1 ml/kg of 0.25% bupivacaine or 0.2% ropivacaine were used to provide intra- and postoperative pain relief. No prophylactic antiemetics were administered. RESULTS: All caudal blocks provided adequate intraoperative pain relief. The incidence of POV was low, 12% at the hospital, 13% for the 24-h study period, and was not affected by age. CONCLUSIONS: We concluded that regional analgesia combined with the avoidance of narcotics administered to children scheduled for elective urologic or lower abdominal procedures, is associated with a lower incidence of POV and that age did not affect the incidence of POV. PMID- 15830768 TI - Post operative analgesia with intrathecal morphine. AB - INTRODUCTION: Sixty eight ASA I and II patients between 18 and 84 years who underwent minor to medium surgery were studied retrospectively. They were divided into 2 groups, on basis on intrathecal morphine administration. MATERIALS AND METHODS: Group I received spinal anesthesia with bupivacaine 12.5-15 mg. Group II received in addition 0.1 mg morphine (preservative free) [MO PF]. Pain assessment, up to 24 hours, was evaluated by the need of salvage medication namely pethidine i.m. (intramuscular). RESULTS: The groups matched for demographic distribution. Twenty one out of 30 patients in Group I needed salvage medication, compared to 6 out of 38 patients in Group II (p < 0.005). Minor complications of intrathecal morphine were noted. These were higher in Group II compared to Group I; itching and pruritus (14 vs. 1); urinary retention (2 vs. 0); nausea and vomiting (1 vs. 0) (p < 0.005). CONCLUSION: This study showed that a single low dose of intrathecal morphine with bupivacaine provides better quality post-operative analgesia than bupivacaine alone, and might offer enough analgesia for minor to medium cases up to 24 hours post-op. However, some side effects have to be taken into consideration. PMID- 15830769 TI - Predictor of sedation during endoscopic retrograde cholangiopancreatography- bispectral index vs clinical assessment. AB - INTRODUCTION: The bispectral index (BIS) has been developed as a monitor of sedation in patients receiving anesthetic and sedative drugs. BIS has been shown to correlate with responsiveness under sedation and anesthesia with a variety of agents. This study was designed to compare BIS with clinical assessment of sedation during ERCP. METHODS: Forty patients were divided into two equal groups: Group I: The sedation level monitored by clinical assessment. Group II: The sedation level monitored by BIS. The patients in both groups did not receive any premedication and the procedure started after sedating the patients with 0.1 mg/kg midazolam and 1 microg/kg fentanyl, additional doses of midazolam were given according to the level of sedation either guided by BIS or clinical assessment. All vital signs and sedation level were monitored, the total dose of sedation was calculated and the rate of recovery and operator's satisfaction rate were evaluated. RESULTS: There was a statistical significant difference between both groups regarding sedative duration, recovery rate, satisfaction rate and the total dose of sedative with non-significant difference in vital signs monitoring. CONCLUSION: These results imply that BIS may be a valuable monitor for safe level of sedation and operator's satisfaction during ERCP. PMID- 15830770 TI - The analgesic effects of epidural diamorphine and levobupivacaine on established lower limb post-amputation stump pain--a comparative study. AB - BACKGROUND AND OBJECTIVES: Pain is necessary for survival but chronic pain is disabling and causes significant health and economic problems. Chronic stump pain (pain localized in the stump for >3 months) after amputation is a significant problem among amputees (5-10%). The mechanism of this phenomenon is not very clear yet. In this study we attempted to better understand the role of peripheral and central mechanisms in this condition. METHODS: 12 patients with established lower limb post-amputation stump pain were given lumbar epidural diamorhpine 5 mg in 20 ml normal saline (NS) and levobupivacaine 0.5% (20 ml) 1 week apart. Baseline pain was recorded and then analgesic and side effects as well as their onset and duration were also assessed. RESULTS: Epidural diamorphine was found to relieve the pain of all patients. Its onset was relatively rapid being started within 5-9 min (median = 5.5) by a smooth sensation of warmth involving the lower trunk and legs. Analgesia was complete in 30 min and had a median duration of 20.5 h. Pruritus in 6 patients was the only side effect due to epidural diamorphine. Epidural levobupivacaine, on the other hand, usually relieved the discomfort but failed to do so in 3 patients only despite adequate neural blockade. Its onset of action was slower (10-20 min, median = 11) and duration of effect (median = 12 h) was also shorter than diamorphine. The effects of levobupivacaine were inferior to diamorphine due to the associated motor and sensory paralyses as well as absence of euphoria. Epidural diamorphine provided profound analgesia with normalization of stump sensations and euphoria, probably due to absorption into the spinal cord causing segmental action. CONCLUSION: While peripheral mechanisms played a role, central mechanisms involving the spinal cord were more important in the modulation of established stump pain in the evaluated patients. PMID- 15830771 TI - The effects of intratracheal dexamethasone on acute lung injury in rabbits- experimental study. AB - BACKGROUND AND OBJECTIVE: The aim of our study was to investigate, in rabbits with hydrochloric (HCl) acid-induced Acute Lung Injury (ALI), the effects of intra-tracheal (i.t.) dexamethasone administration on neutrophil and platelet counts, plasma malonyl dialdehyde (MDA) level, histopathology, and arterial blood gases. METHODS: Twenty-eight New Zealand rabbits were randomly divided into a control (n = 14) and dexamethasone groups (n = 14). Anesthesia was applied with ketamine (50 mg kg(-1) h(-1), i.m.) and tracheostomy was performed. Both groups received 2 ml kg(-1) HCl (hydrochloric acid) i.t.. Five minutes later, 0.9% saline was given to the controls while the other group received i.t. dexamethasone. MDA levels of the plasma, neutrophil and platelet counts, and arterial blood gases were recorded at the beginning of the study and then at two and five hours. The rabbits were ventilated in pressure control mode for 5 hours. At the end of the study, the lungs were examined by light microscopy and the changes evaluated on a scale of 0 to 4. RESULTS: Neutrophil and platelets counts (p < 0.01) and PaO2 (p < 0.05) were greater in the dexamethasone group than the controls at five hours. Plasma MDA level (p < 0.01) and PaCO2 (p < 0.01) were greater in the control group. Histopathological changes were less severe in the dexamethasone group. CONCLUSION: Giving dexamethasone i.t. may have beneficial effects on the development of ALI. PMID- 15830772 TI - Successful management of intra-operative cardiac arrest in a child for pheochromocytoma resection--a case report. PMID- 15830773 TI - Quality improvement in anesthetic practice--incidence of sore throat after using small tracheal tube. AB - BACKGROUND AND OBJECTIVE: Sore throat following surgery is common and is due to multitude of factors. The highest incidence of sore throat tends to occur in patients who have undergone tracheal intubation. Between 14.4% to 50% of intubated patients complain of sore throat and hoarseness in the immediate postoperative period, 3% of them are still hoarse after a week. This contributes to higher incidence of postoperative morbidity and patient dissatisfaction of the service. A prospective study was conducted to demonstrate the incidence of hoarseness and sore throat following the use of small tracheal tubes. METHODS: Adult patients, 16 years old and above, presented for tonsillectomy, nasal surgery and/or for functional endoscopic sinus surgery (FESS) were included in the study. Patients who had signs of, common cold were excluded from the study. Size 7-7.5 mm cuffed tracheal tube was used in males and size 6-6.5 mm in females. Patients were asked during 24 hours postoperatively about the presence of hoarseness of voice and of their satisfaction of the service. RESULTS: There were 1618 patients 883 males and 735 females, ages: 16-62 yrs included in the study between February 2000 and end of May 2003. 189 (11.7%) developed postoperative hoarseness of voice and 1429 (88.3%) did not have hoarseness or sore throat (P = 0.0001). Patient satisfaction was 95%. CONCLUSIONS: The use of small tube in intubating the trachea, together with other measures such as lubricating the tube with water soluble jelly, careful airway instrumentation, intubation only when patient was fully relaxed, careful suctioning technique, and extubation when the tracheal tube cuff was fully deflated, have dramatic effects on minimizing the incidence of postoperative hoarseness and sore throat. Accordingly, patient satisfaction has been reported to be high. PMID- 15830774 TI - Intensive care of patients with the syndrome of hemolysis elevated liver enzymes and low platelets (HELLP). PMID- 15830775 TI - CABG under thoracic and lumbar epidural block in a conscious patient--a case report. AB - In summary, a coronary artery bypass graft was carried out on an awake patient with the aid of lumbar and thoracic epidural anesthesia, due to dissection of the saphenous graft from the lower limb. The procedure was done using an off pump technique and the patient required no extra sedation of analgesia. In conclusion, we think that many high risk patients could be treated using this technique and that awake CABG seems to be a promising adjunct to current minimally invasive CABG techniques. PMID- 15830776 TI - Endobronchial hemorrhage following endobronchial intubation--a case report. PMID- 15830777 TI - "Reducing cardiovascular responses to laryngoscopy and tracheal intubation: a comparison of equipotent doses of tramadol, nalbuphine and pethidine, with placebo". PMID- 15830778 TI - Radiation protection in deep space. PMID- 15830779 TI - [Oblique incidence of electron beams--comparisons between calculated and measured dose distributions]. AB - Clinical applications of high-energy electron beams, for example for the irradiation of internal mammary lymph nodes, can lead to oblique incidence of the beams. It is well known that oblique incidence of electron beams can alter the depth dose distribution as well as the specific dose per monitor unit. The dose per monitor unit is the absorbed dose in a point of interest of a beam, which is reached with a specific dose monitor value (DIN 6814-8). Dose distribution and dose per monitor unit at oblique incidence were measured with a small-volume thimble chamber in a water phantom, and compared to both normal incidence and calculations of the Helax TMS 6.1 treatment planning system. At 4 MeV and 60 degrees, the maximum measured dose per monitor unit at oblique incidence was decreased up to 11%, whereas at 18 MeV and 60 degrees this was increased up to 15% compared to normal incidence. Comparisons of measured and calculated dose distributions showed that the predicted dose at shallow depths is usually higher than the measured one, whereas it is smaller at depths beyond the depth of maximum dose. On the basis of the results of these comparisons, normalization depths and correction factors for the dose monitor value were suggested to correct the calculations of the dose per monitor unit. PMID- 15830780 TI - [TierSPECT: performance of a dedicated small-animal-SPECT camera and first in vivo measurements]. AB - This paper presents the performance of a new small-animal camera (TierSPECT) devised for the in vivo measurements of radiolabeled substances in small laboratory animals such as mice and rats. In a scatter medium, the camera has a tomographic spatial resolution of 2.87 mm and a sensitivity of 22 cps/MBq in a usable Field-of-View (FOV) with a diameter of 82 mm. The planar homogeneity amounts to 3.3%, the tomographic homogeneity lies between 3.2% and 3.5%. The deviation between filled and measured concentration of activity in a cylindrical 4-chamber-phantom was smaller than 2.6%. Using a novel rat head phantom with chamber volumes in the order of magnitude of the spatial resolution (between 0.065 ml and 0.19 ml) it could be demonstrated that studies of the rat neostriatal dopaminergic system are feasible under observance of physiological conditions. In vivo studies using [99mTc]diphosphonato-1,2-propandicarbonic acid (99mTc-DPD) and [123I]N-omega-fluoropropyl-2beta-carbomethoxy-3beta-(4 iodophenyl)-nortropane (123I-FP-CIT) proved that bone metabolism and dopamine transporter binding can be visualized with the TierSPECT. The fusion of 99mTc-DPD and 123I-FP-CIT images allowed the differentiation between intra- and extracerebral structures. Pretreatment with methylphenidate resulted in blockade of striatal dopamine transporter binding. PMID- 15830781 TI - [Investigations of beta-dosimetry at two different sources for the cardiovascular brachytherapy]. AB - The intracoronary brachytherapy is used at the Hamburg University Hospital as a method to treat in-stent restenosis. Two different radiochromic film types were applied to obtain dosimetric information of the beta-sources used (32P and 90Sr/90Y). First, these films were analyzed for their suitability for dosimetry. Within the investigated dose range (MD-55-2: 0 to 33 Gy, HD-810: 0 to 105 Gy), both films showed a linear behavior between the dose and the optical density (OD). Because radiochromic films are subject to time-based changes in OD, a method for colour stabilization was investigated (RCS-method). This method allowed to greatly shorten the time between irradiation and evaluation from 24 hours (time necessary for the film to reach a quasi-stable status) to 2.5 hours. Colour-stabilized films can also be stored for a long time and reanalyzed with almost the same results. Within the limits of the measurements error, both film types showed an energy independent response. Within the dose profiles, analyses of the two source types resulted in differences of 13.5% (32P) and 21% (90Sr/90Y). These inhomogenities are consistent with the fabrication tolerances given by the manufactures. PMID- 15830782 TI - A simple dose-response relationship for modeling secondary cancer incidence after radiotherapy. AB - Estimates of secondary cancer risk after radiotherapy are becoming more important for comparative treatment planning. There is great uncertainty concerning the dose-response relationship for radiation-induced carcinogenesis at doses higher than 4 Gy. The purpose of this report is to determine a simple dose-response relationship for secondary cancer incidence after radiotherapy treatment which can be used for comparative treatment planning. In this report a simple one parameter model to estimate the complication probability of secondary cancer was fitted to literature data on secondary cancer incidence after radiotherapy. The results showed a linear dose-response relationship in the low-dose part and an exponentially decreasing one after a maximum at around 10 Gy. The observed dose response relationship and the literature data used to fit the dose-response indicate that cell death effects are important for the explanation of secondary cancer incidence. Even using a dose and dose-rate effectiveness factor (DDREF) of two (instead of one), a cancer incidence maximum is observed at around 10 Gy, with decreasing incidence at higher doses. PMID- 15830783 TI - [Measurements of respiratory motion using fast magnetic resonance imaging and inductively-coupled marker coils]. AB - The respiratory motion of the thoracic wall provides indirect information about the breathing displacement of the inner organs. To analyze the correlation between thoracic wall and lung motion for applications in radiation therapy, the breathing displacement of the lung is visualized with a fast gradient echo pulse sequence (trueFISP) at a rate of 2-3 images/sec. For quantification of the motion, a small inductively-coupled marker coil is attached to the chest wall and detected with a fast projection technique. Since the marker coil generates a flip angle amplification (factor 15) in its interior, very small nominal flip angles of 2 degrees can be used during the projection measurements which do not affect the image quality of the trueFISP images. Volunteer studies with the marker coil showed a good agreement with simultaneously acquired breathing belt data and position information extracted from the MR images. Whereas the breathing belt provided reliable data only within a certain dynamic range, the marker coil could detect also extreme breathing excursions with a precision better than 2 millimeters. PMID- 15830784 TI - Collagene order of articular cartilage by clinical magnetic resonance images and its age dependency. AB - The present paper describes a novel method to obtain information on the degree of order of the collagen network of the knee meniscal cartilage by means of a single clinical MRI. Images were obtained from 34 healthy volunteers aged between 6 and 76 years as well as from one patient with clinically-diagnosed arthrosis at the age of 32 and 37 years. A Siemens Vision (1.5 T) MRT with TR=750 ms, TE=50 ms, FoV=160 mm, and Matrix 512x512 was used for this purpose. The MR signal intensities of the cartilage were read out along slices with constant height above the subchondral bone and plotted versus the actual angle to the external magnetic field. The obtained intensity curves were fitted by a model distribution, and the degree of order of the collagen fibers was calculated. For the knee meniscal cartilage, there was an age-dependency of the degree of order and a significant deviation of the volunteer with arthrosis from the normal curve. The results are discussed in view of the arcade model and of a possible use of non-invasive clinical MRT for the detection of early arthrotic changes of cartilage. PMID- 15830785 TI - [Increased fragmentation efficiency by enhancement of cavitation for extracorporal shock wave lithotripsy]. AB - The non-invasive disintegration of kidney stones using shock waves, referred to as extracorporal shock wave lithotripsy, has been successful for more than twenty years in treating patients having renal and ureteral stones. Two modified shock wave generators are described in this article. The novel systems produce two similar shock waves (tandem shock waves) generated with a short time delay. The second shock wave arrives during collapse of the bubbles generated in the neighborhood of the stone due to the first shock wave. This may increase cavitation bubble collapse and could enhance cavitation-induced damage to kidney stones during shock wave lithotripsy. In vitro comparison of standard systems with the new designs showed that fragmentation efficiency of artificial kidney stones was significantly enhanced using tandem shock waves. PMID- 15830786 TI - [Pioneers of medical physics--Theophil Christen (1873-1920)]. AB - The Swiss mathematician and physician Theophil Christen was one of the first important pioneers in Medical Physics. He structured the previously confused field of physical concepts and definitions for medical applications of X-rays and paved the way for the modern understanding of dosimetry. PMID- 15830787 TI - [Pertussis: its evolution in Argentina at the end of the twentieth century]. AB - The objective of this study was to analize data of pertussis morbidity and mortality in Argentina and vaccination coverage with combined pertussis vaccine (DwPT), which was available from 1969 until 2000 inclusive, and to investigate their interrelation. A retrospective study was performed based on National Registers. Notification and mortality annual mean variation rates and geometric mean rates, by periods, were analyzed, and correlation coefficients between each one of them and with vaccination coverage were calculated. Notification annual mean variation rates for 1969-2000 were -14.34 and for 1980-2000 -17.26; mortality annual mean variation rates for 1980-2000 was -10.41. Vaccination coverage in infants up to one year of age (3 doses) was 44.4% in 1980; less than 60% until 1982 and higher than 80% after 1990. A highly significant inverse correlation was observed, between (a) vaccine implementation, increase in its coverage, introduction of a fifth vaccine dose, and (b) notification and mortality rates. It was observed during the study, that in our country there is a relatively little knowledge about disease occurrence in adults, and an apparent absence of population-based studies performed on the efficacy of erythromycin chemo prophylaxis in epidemics outbreaks. Based on these data, strategies for a better surveillance and control of pertussis, are exposed. PMID- 15830788 TI - [Maternal morbidity and perinatal mortality in HELLP syndrome. Multicentric studies in intensive care units in Buenos Aires area]. AB - We analyzed the clinical characteristics, complications, severity, and maternal and fetal survival of patients suffering from HELLP syndrome (Hemolysis, Elevated Liver enzymes level, Low Platelet count) requiring admission to the intensive care unit in four hospitals from Buenos Aires area, Argentina. Data was revised in the charts from March 1997 to March 2003 and 62 patients were included in the study. During the second half of pregnancy or immediate puerperal period, diagnostic criteria were defined on the basis of preeclampsia and the following laboratory abnormalities: platelet count nadir <150,000/mm3, serum hepatic aminotransferases >70 UI/l, and serum lactic dehydrogenase >600 UI/l, total bilirubin >1.2 mg/dl and/or periferical blood smear with hemolysis. The mean maternal age was 28 +/- 8 years; parity 2.7 +/- 2.3; gestational age 33 +/- 4 weeks. According to platelet count, 23 cases were identified to class 1, 29 to class 2 and the rest to Martin's class 3. There were 16 eclamptic patients. The platelet count was 67,604 +/- 31,535/mm3; alanine aminotransferase 271 +/- 297 UI/l; aspartate aminotransferase 209 +/- 178 UI/l; serum lactic dehydrogenase 1444 +/- 1295 UI/l; serum creatininine levels 1.1 +/- 0.8 mg/dl. Forty-one patients had diverse degree of renal function damage, renal dialysis and plasmapheresis was required in one female. Respiratory failure due to pulmonary edema was observed in four patients. All obstetric patients survived. There were four perinatal deaths. In our population sample, low rate of life-threatening maternal complications and low perinatal mortality were observed. PMID- 15830789 TI - [Use of infliximab in patients of a rheumatologic center]. AB - The objective of this study was to obtain post-marketing information about the use of infliximab in an ambulatory setting. We studied--retrospectively and prospectively--the case records of patients with rheumatoid arthritis (n=37), psoriatic arthritis (n=5), mixed connective tissue disease (n=1), and ankylosing spondylitis (n=2) who received infliximab (3 mg/kg) from August 2000 to January 2003. Descriptive values were given as percentage, mean or median, and standard deviation or interquartile range. Wilcoxon test was used for paired analysis of pre/post doses of corticosteroids, non-steroidal anti-inflammatory drugs, and methotrexate therapy. A p value < or = 0.05 was considered significant. Forty five patients were included. A total of 207 infusions were administered. In 4 patients the treatment was permanently discontinued due to severe back pain during the infusion (2 cases) and serious anaphylactic reactions (2 cases). Other adverse reactions occurring during infusions were mild and successfully managed with standard treatment. A case of staphylococcal septic arthritis resolved with standard antibiotic treatment. No patient had evidence of active tuberculosis. One patient with rheumatoid arthritis and chronic renal insufficiency, received treatment with infliximab 1.9 mg/kg, every 30 days, with no changes in renal function. Due to improvement of symptoms, 14/39 (35.9%) patients could decrease the doses of corticosteroids, 15/43 (34.8%) decreased the doses of antiinflammatory drugs and 12/34 (35.3%) decreased methotrexate dosage. Although some questions remain to be elucidated, this case series shows the drug safety profile, the possibility to reduce concomitant drug doses, as well as individual approaches for situations where there are not yet guidelines available, so that rheumatologists have to make decisions based on clinical needs. PMID- 15830790 TI - Overestimation of thoracic gas volume during the airway resistance maneuver. A potential error in the diagnosis of air trapping. AB - There are no data published about the agreement between the measurement of thoracic gas volume (TGV) during the airway resistance (TGV-Raw) and the conventional technique described by Dubois. The aim of this study was to establish the agreement between both methods to measure TGV. We studied eighty consecutive subjects. Only sixty-six performed acceptable plethysmography maneuvers. The patients were measured with a constant volume plethysmograph (Medical Graphics 1085 DL). TGV was performed in the same patient with two techniques: 1) during the airway resistance (Raw) measurement (TGV-Raw) and 2) during quiet breathing at the end of expiration (TGV). The panting frequency was 1 to 2 Hz with both maneuvers. The differences between both techniques were expressed in percentage (deltaTGV %) and absolute values (deltaTGV). The TGV-Raw of the whole group was higher than TGV (3.69 +/- 1.08 l vs 3.28 +/- 1.05 l, p < 0.001). Similarly, the subgroups of patients had a greater TGV-Raw than TGV (Normal: 3.44 +/- 0.77 l vs 2.98 +/- 0.72 l , p < 0.001; Obstructive: 4.08 +/- 1.19 l vs 3.71 +/- 1.15 l, p < 0.001; Restrictive: 2.62 +/- 0.49 l vs 2.25 +/- 0.51 l, p < 0.01). There was a considerable lack of agreement between the TGV-Raw and TGV, with discrepancies of up to +0.95 l or +34%. The deltaTGV % was similar between the patients' subgroups and between the subjects with different degree of airflow obstruction (Normal: 16.5 +/- 10%, Obstructive: 10.8 +/- 9.4%, Restrictive: 18 +/- 14.3%, p NS; mild obstruction: 10.7 +/- 11%, moderate obstruction: 12.3 +/- 5.7, severe obstruction: 10.1+/- 6.6, p NS). In conclusion, TGV-Raw was larger than TGV. This was because the patients generally panted at a volume above FRC when performing the TGV-Raw maneuver. TGV-Raw should not be used to estimate FRC because FRC would be overestimated and the diagnosis of air trapping may be erroneous. PMID- 15830791 TI - [Severe acute respiratory syndrome. The public health laboratory in a global emergency]. AB - By the end of year 2002 there was an outbreak of atypical pneumonia in Southeast Asia which soon spread to other continents. This new severe acute respiratory syndrome (SARS) was produced by a novel coronavirus. Due to the severity of the situation and risk of introduction of this pathology in our country, the need to arrange specific laboratory diagnostic tests arose. Classic techniques, such as the electron microscopy and molecular biology test such as retrotranscription followed by the polymerase chain reaction (RT-PCR) were implemented. The araldit included cells infected with bovine coronavirus which allowed the viral particles to be visualized easily but it took more time in comparison with the negative staining of free particles from viral cultures. RT-PCR was able to detect RNA of isolated viruses from cases in Hong Kong and Germany. PMID- 15830792 TI - [Detection of a non-standard mutation in the ret protoncogene by site directed mutagenesis]. AB - MEN2A is an autosomic dominant disease, characterized by medullary thyroid cancer, pheochromocytoma and parathyroid hyperplasia. Mutations in the ret proto oncogene are associated with this disease, with almost 100% of penetrance. The gene, situated on chromosome 10q11.2, codes for a transmembrane protein with a tyrosinkinase-like receptor function. Mutations that affect its extracellular domain, stimulate spontaneous homodimerization and elevate the basal tyrosinkinase activity. The codon 634 of the gene is considered a hot-spot site, since it is mutated in 85% of the MEN2A families. Our group developed in 2002 an indirect and costless strategy to detect alterations in this site. We present a family suspected of having MEN2A. We applied our PCR based indirect strategy on the DNA of the index patient and found that there was no mutation in that site. Posterior sequencing of exon 10 and 11 confirmed that the mutation affecting this family was in codon 611. Thus, we developed a new costless family-specific strategy based on mutagenic PCR and enzymatic cuts to diagnose all the family members. A seven-year old boy with this mutation was preventively thyroidectomized. In this way, combining the indirect methodology for codon 634 previously developed by our group, and a posterior family-specific mutation detection strategy, we were able to diagnose and intervene presymptomatically the family members, avoiding sending all the samples to foreign centers. PMID- 15830793 TI - [Acquired palmoplantar keratoderma with hypothyroidism]. AB - We report the case of a 45 year-old male with palmoplantar hyperkeratosis. He had chronic thyroiditis with severe hypothyroidism. Skin lesions improved upon thyroid hormone replacement. Complete resolution of palmoplantar keratoderma was achieved after 7 months of therapy. The association of acquired palmoplantar keratoderma with hypothyroidism even though extremely unusual, should be considered in the differential diagnosis. It is a reversible condition. PMID- 15830794 TI - [Left ocular proptosis and obstruction of upper airways in a cavum lymphoma]. PMID- 15830795 TI - [Regulatory cytokines in the response to the allogeneic renal transplant]. AB - The outcome of the kidney allograft mainly depends on the immune response and on its complex regulation, where the cytokine network and other mediators play an important role. At present, kidney biopsy is the most useful tool for monitoring the transplant rejection and the diagnosis of the associated nephropathies, in spite of the invasiveness of the procedure. Thus, it is of great interest to find alternative tools for diagnosis. The evaluation of regulatory cytokines is a simple procedure of low cost that could be useful to increase the sensitivity of the detection of polymorphic differences, to predict the graft acceptance and for the early detection of rejection. Recent studies suggest that the high production of pro-inflammatory mediators, such as Th1 cytokines, could be detrimental, whereas the production of anti-inflammatory regulatory cytokines, such as interleukin (IL)-10 and tumor necrosis factor (TGF)-beta, could be beneficial for graft survival. In the early stages, the cellular cytotoxicity is activated by the Th1 response and the detection of cytotoxic molecules is associated to the acute rejection. Later, the balance between pro and anti-inflammatory mediators and the regulation of their levels could be more important. In this regard, TGF beta is also fibrogenic and a high local production can contribute to kidney damage. On the other hand, the increased production of IL-10 in response to the allogeneic stimuli could be, in most cases, an important marker of long-term acceptance. PMID- 15830796 TI - [Importance of three triatoma vectors of Trypanosoma cruzi in Mexico]. AB - Observations made in the field and in the laboratory on three vectors of Trypanosoma cruzi: Triatoma barberi, T. pallidipennis and T. dimidiata are here presented. Results are compared in relation with their ecotopes, vectorial capacity, biological characteristic, entomological indexes and capture places. In relation with their ecotopos T. barberi has preference for the interior of the human home, in walls, directly in contact with beds; T. pallidipennis, when it is inside the houses, prefers floors and among clothes, although their main ecotope is wild; T. dimidiata has localization in floors, specially under the beds, in the angle between wall and floor; T. barberi and T. dimidiata are attracted by the light. The cycles of life were of 523, 171 and 510 days, respectively for Triatoma barberi, T. pallidipennis and T. dimidiata. The vectorial capacity was determined according to the time taken in defecating during or after its sucking blood: T. barberi defecates during its feeding, while T. pallidipennis and T. dimidiata make it from 10 to 20 and of 20 to 30 minutes after starting the process, respectively. The indexes of natural infection were of 56.6 for T. barberi, 29 for T. pallidipennis and 15.6 for T. dimidiata, The metacyclogenics indexes that indicate percentages of metacyclics trypomastigotes in later intestine of vectors was in T. barberi 76.6, T. pallidipennis 15 and T. dimidiata 26, with what it can be concluded that the first species is the best transmitter of T. cruzi in Mexico. PMID- 15830797 TI - [Remembering Alfredo Lanari (1910-1985) by his editorials]. PMID- 15830798 TI - [Mitochondrias and Janus green]. PMID- 15830799 TI - [Antiretroviral (ARV) therapy. Advances and deficits left by the year 2004]. PMID- 15830800 TI - [Treatment of Chagas disease with benznidazole and thioctic acid]. PMID- 15830801 TI - [Hypothesis: An alternative pathway for the regulation of inflammation. Hormesis and inflammation]. PMID- 15830802 TI - Biofilm formation of Pseudomonas putida IsoF: the role of quorum sensing as assessed by proteomics. AB - Pseudomonas putida strains are frequently isolated from the rhizosphere of plants and many strains promote plant-growth, exhibit antagonistic activities against plant pathogens and have the capacity to degrade pollutants. Factors that appear to contribute to the rhizosphere fitness are the ability of the organism to form biofilms and the utilization of cell-to-cell-communication systems (quorum sensing, QS) to co-ordinate the expression of certain phenotypes in a cell density dependent manner. Recently, the ppu QS locus of the tomato rhizosphere isolate P. putida Iso F was characterized and an isogenic QS-negative ppuI mutant P. putida F117 was generated. In the present study we investigated the impact of QS and biofilm formation on the protein profile of surface-associated proteins of P. putida IsoF. This was accomplished by comparative proteome analyses of the P. putida wild type IsoF and the QS-deficient mutant F117 grown either in planktonic cultures or in 60 h old mature biofilms. Differentially expressed proteins were identified by peptide mass fingerprinting and database search in the completed P. putida KT2440 genome sequence. The sessile life style affected 129 out of 496 surface proteins, suggesting that a significant fraction of the bacterial genome is involved in biofilm physiology. In surface-attached cells 53 out of 484 protein spots were controlled by the QS system, emphasizing its importance as global regulator of gene expression in P. putida IsoF. Most interestingly, the impact of QS was dependent on whether cells were grown on a surface or in suspension; about 50% of the QS-controlled proteins identified in planktonic cultures were found to be oppositely regulated when the cells were grown as biofilms. Fifty-seven percent of all identified surface-controlled proteins were also regulated by the ppu QS system. In conclusion, our data provide strong evidence that the set of QS-regulated proteins overlaps substantially with the set of proteins differentially expressed in sessile cells. PMID- 15830803 TI - Description of four novel species of Xenorhabdus, family Enterobacteriaceae: Xenorhabdus budapestensis sp. nov., Xenorhabdus ehlersii sp. nov., Xenorhabdus innexi sp. nov., and Xenorhabdus szentirmaii sp. nov. AB - The taxonomic affiliation was determined for four Xenorhabdus strains isolated from four Steinernema hosts from different countries. As compared to the five validly described Xenorhabdus species, i.e., X. nematophila, X. japonica, X. beddingii, X. bovienii and X. poinarii, these isolates represented novel species on the basis of 16S rRNA gene sequences and riboprint patterns, as well as by physiological and metabolic properties. They were named Xenorhabdus budapestensis sp. nov., type strain DSM 16342T, isolated from Steinernema bicornutam; Xenorhabdus ehlersii sp. nov., type strain DSM 16337T, isolated from Steinernema serratum; Xenorhabdus innexi sp. nov., type strain DSM 16336T isolated from Steinernema scapterisci; and Xenorhabdus szentirmaii sp. nov., type strain DSM 16338T, isolated from Steinernema rarum. PMID- 15830804 TI - Erythrobacter vulgaris sp. nov., a novel organism isolated from the marine invertebrates. AB - Four yellow-pigmented, gram-negative, chemoorganotrophic aerobic bacteria were isolated from starfish Stellaster equestris (strains 022-2-10T, 022-2-9, and 022 2-12) and soft coral (unidentified species) (strain 022-4-7) collected in the South China Sea. 16S rRNA gene sequence-based analyses of the new organisms revealed that Erythrobacter spp. were the closest relatives and shared the highest similarity of 98.7% to E. citreus, 98.5% to E. flavus, 97.9% to E. litoralis and 97.6% to E. longus. The novel organisms were tolerant to 3-6% NaCl, grew between 10 degrees C and 40 degrees C, and were not able to degrade gelatin, casein, and agar, while degraded Tween 80. Two strains (022-2-9 and 022-2-12) could weakly degrade starch. All strains produced a large pool of carotenoids and did not have Bacteriochlorophyll a. Phosphatidylethanolamine (30-36%), phosphatidylglycerol (39-46%), and phosphatidylcholine (21-27%) were the predominant phospholipids. Sphingoglycolipid was not detected. The major fatty acids were 16:0 (6-11%), 16:1omega7 (12-15%), and 18:1omega7 (46-49%). The two hydroxy fatty acids, 13:0-2OH, 14:0-2OH, 15:0-2OH, 16:0-2OH were also present. The G + C content of the DNAs ranged from 61 to 62 mol%. The level of DNA similarity among four strains was conspecific and ranged from 94% to 98%. Even though new strains and other species of the genus had rather high level of 16S rRNA gene sequence similarities, DNA-DNA hybridization experiments showed only 33 39% of binding with the DNA of the type strains. On the basis of these results and the significant differences demonstrated in the phenotypic and chemotaxonomic characteristics, it is suggested that the new organisms be classified as a novel species; the name Erythrobacter vulgaris sp. nov. is proposed. The type strain is 022-2-10T (= KMM 3465T = CIP 107841T). PMID- 15830805 TI - Oceanimonas smirnovii sp. nov., a novel organism isolated from the Black Sea. AB - A slightly creamy, melanogenic, gram-negative, aerobic bacterium was isolated from seawater sample collected in the Karadag Natural Reserve of the Eastern Crimea, the Black Sea. The novel organism was chemoorganotrophic, had no obligate requirement in NaCl, tolerated to 12% NaCl, grew between 10 and 45 degrees C, was slightly alkaliphilic, and was not able to degrade starch, gelatin, agar, and Tween 80. 16S rRNA gene sequence-based analyses of the new organism revealed that Oceanimonas doudoroffii ATCC 27123T, Oceanimonas baumanii ATCC 700832T, and Oceanisphaera litoralis DSM 15406T were the closest relatives (similarity around 97%-96%). The G + C content of the DNA of the strain 31-13T was 55.5mol%. Phosphatidylethanolamine (49.0%), phosphatidylglycerol (41.8%), and diphosphatidylglycerol (9.2%) were the predominant phospholipids. The major fatty acids were 16:0 (24.1%), 16:1omega7 (40.3%), and 18:1omega7 (29.2%). On the basis of the significant differences demonstrated in the phenotypic and chemotaxonomic characteristics, it is suggested that the bacterium be classified as a novel species; the name Oceanimonas smirnovii sp. nov. is proposed. The type strain is 31-13T (UCM B-11076T = LMG 22147T = ATCC BAA-899T). PMID- 15830806 TI - Pseudoxanthomonas kaohsiungensis, sp. nov., a novel bacterium isolated from oil polluted site produces extracellular surface activity. AB - During screening for biosurfactant-producing bacteria, a strain designated J36T was isolated from oil-polluted site near Kaohsiung city located in southern Taiwan. Cells of this organism were gram-negative rods motile by means of a single polar flagellum. Strain J36T grew well in complex media under optimum conditions of 35 degrees C and pH 7. The extracellular products of the strain expressed emulsification activity. During cultivation on olive oil as the sole carbon and energy source, the culture supernatant of strain J36T reduced surface tension of the medium from 68 to 32.6 dyne/cm. The 16S rRNA gene sequence analysis indicates that strain J36T is a member of Xanthomonas group within the gamma-Proteobacteria. The organism belongs to the genus Pseudoxanthomonas and represents a novel species within this genus according to phylogenetic analysis of 16S rDNA sequences, DNA-DNA similarity data, whole-cell protein analysis, physiological and biochemical characteristics, as well as fatty acid compositions. The predominant cellular fatty acids of strain J36T were 15:0 iso (about 26%), 17:1 iso omega9c (about 25%), and 15:0 anteiso (about 10%). Its DNA base ratio was 60.1 mol% G+C. We propose to classify strain J36T (= BCRC 17375T = LMG 22530T) as Pseudoxanthomonas kaohsiungensis sp. nov. PMID- 15830807 TI - Marinomonas aquamarina sp. nov., isolated from oysters and seawater. AB - The characterization of three bacterial strains isolated from cultured oysters and seawater at the Spanish Mediterranean coast has been performed. Strains were phenotypically and genetically characterized and the results led us to identify them as members of the genus Marinomonas. A phylogenetic analysis based on the almost complete 16S rDNA sequences clustered all three strains together (with sequence similarities around 99.8%) in the vicinity of M. communis and M. vaga sequences and distantly related to the other four species of the genus. The most closely related species was M. communis that shared 97.4-97.6% with the Mediterranean strains. DNA-DNA hybridizations were performed to clarify the taxonomic position of our isolates and the results confirmed their specific isolation, with interspecific binding ratios below 59%. We propose the bacteria to constitute a new Marinomonas species, i.e. M. aquamarina and strain 11SM4T (CECT 5080T, CIP 108405T, CCUG 49439T) as the type strain. PMID- 15830808 TI - Carnobacterium divergens and Carnobacterium maltaromaticum as spoilers or protective cultures in meat and seafood: phenotypic and genotypic characterization. AB - Carnobacterium, a genus of lactic acid bacteria, frequently dominate the microflora of chilled vacuum- or modified atmosphere-packed meat and seafood. In this study Carnobacterium isolates were characterized by phenotypic and molecular methods in order to investigate the association of species and intra-species groups with distinct kinds of meat and seafood. Of 120 test strains, 50 originated from meat (beef and pork products, including 44 strains isolated during this study and 6 strains obtained from culture collections) and 52 from seafoods (cod, halibut, salmon, shrimps and roe products). In addition, 9 reference strains of Carnobacterium spp from other sources than meat and fish and 9 reference strains of lactic acid bacteria belonging to other genera than Carnobacterium were included. Numerical taxonomy relying on classical biochemical reactions, carbohydrate fermentation and inhibition tests (temperature, salt, pH, chemical preservatives, antibiotics, bacteriocins), SDS-PAGE electrophoresis of whole cell proteins, plasmid profiling, intergenic spacer region (ISR) analysis and examination of amplified-fragment length polymorphism (AFLP) were employed to characterize the strains. The numerical taxonomic approach divided the carnobacteria strains into 24 groups that shared less than 89% similarity. These groups were identified as Carnobacterium divergens with one major cluster (40 strains) and 7 branches of one to four strains, Carnobacterium maltaromaticum (previous C. piscicola) with one major cluster (37 strains) and 9 branches of one to four strains and Carnobacterium mobile (three branches consisting in total of 4 strains). Branches consisting of references strains of the remaining Carnobacterium spp. were separated from clusters and branches of C. divergens, C. maltaromaticum and C. mobile. Isolates from the main clusters of C. divergens and C. maltaromaticum were found both in fresh and lightly preserved meat and seafood products. High phenotypic intra-species variability was observed for C. divergens and C. maltaromaticum but despite this heterogeneity in phenotypic traits a reliable identification to species levels was obtained by SDS-PAGE electrophoresis of whole cell proteins and by ISR based on 16S-23S rDNA intergenic spacer region polymorphism. With AFLP, two distinct clusters were observed for C. divergens but only one for C. maltaromaticum. The two C. divergens clusters were not identical to any of the clusters observed by numerical taxonomy. A limited number of C. divergens and C. maltaromaticum isolates possessed a biopreservative potential due to their production of bacteriocins with a wide inhibition spectrum. This study serves as a base-line for further investigations on the potential role of species of Carnobacterium in foods where they predominate the spoilage microflora. PMID- 15830810 TI - Community analysis of a full-scale anaerobic bioreactor treating paper mill wastewater. AB - To get insight into the microbial community of an Upflow Anaerobic Sludge Blanket reactor treating paper mill wastewater, conventional microbiological methods were combined with 16S rRNA gene analyses. Particular attention was paid to microorganisms able to degrade propionate or butyrate in the presence or absence of sulphate. Serial enrichment dilutions allowed estimating the number of microorganisms per ml sludge that could use butyrate with or without sulphate (10(5)), propionate without sulphate (10(6)), or propionate and sulphate (10(8)). Quantitative RNA dot-blot hybridisation indicated that Archaea were two-times more abundant in the microbial community of anaerobic sludge than Bacteria. The microbial community composition was further characterised by 16S rRNA-gene targeted Denaturing Gradient Gel Electrophoresis (DGGE) fingerprinting, and via cloning and sequencing of dominant amplicons from the bacterial and archaeal patterns. Most of the nearly full length (approximately 1.45 kb) bacterial 16S rRNA gene sequences showed less than 97% similarity to sequences present in public databases, in contrast to the archaeal clones (approximately. 1.3 kb) that were highly similar to known sequences. While Methanosaeta was found as the most abundant genus, also Crenarchaeote-relatives were identified. The microbial community was relatively stable over a period of 3 years (samples taken in July 1999, May 2001, March 2002 and June 2002) as indicated by the high similarity index calculated from DGGE profiles (81.9+/-2.7% for Bacteria and 75.1+/-3.1% for Archaea). 16S rRNA gene sequence analysis indicated the presence of unknown and yet uncultured microorganisms, but also showed that known sulphate-reducing bacteria and syntrophic fatty acid-oxidising microorganisms dominated the enrichments. PMID- 15830811 TI - Protein transport and trafficking in Plasmodium falciparum-infected erythrocytes. AB - The human malarial parasite Plasmodium falciparum extensively modifies its host erythrocyte, and to this end, is faced with an interesting challenge. It must not only sort proteins to common organelles such as endoplasmic reticulum, Golgi and mitochondria, but also target proteins across the 'extracellular' cytosol of its host cell. Furthermore, as a member of the phylum Apicomplexa, the parasite has to sort proteins to novel organelles such as the apicoplast, micronemes and rhoptries. In order to overcome these difficulties, the parasite has created a novel secretory system, which has been characterized in ever-increasing detail in the past decade. Along with the 'hardware' for a secretory system, the parasite also needs to 'program' proteins to enable high fidelity sorting to their correct subcellular location. The nature of these sorting signals has remained until relatively recently, enigmatic. Experimental work has now begun to dissect the sorting signals responsible for correct subcellular targeting of parasite-encoded proteins. In this review we summarize the current understanding of such signals, and comment on their role in protein sorting in this organism, which may become a model for the study of novel protein trafficking mechanisms. PMID- 15830809 TI - Nocardioides aromaticivorans sp. nov., a dibenzofuran-degrading bacterium isolated from dioxin-polluted environments. AB - Seven strains of dibenzofuran (DF)-degrading bacteria isolated from dioxin polluted environments were characterized. These isolates were able to grow with dibenzofuran as the sole carbon and energy source. During the growth with dibenzofuran, they produced a soluble yellow metabolite that exhibited a unique pH-dependent shift of absorption maxima. Dibenzo-p-dioxin and biphenyl were also degraded with pigment production. The isolates were strictly aerobic and chemoorganotrophic and had gram-positive, nonmotile, rod-shaped cells. Chemotaxonomic analyses showed that cells contained L,L-diaminopimeric acid in the peptidoglycan, branched-chain fatty acids as major fatty acids, and menaquinone MK-8(H4) as the sole respiratory quinone. The G + C content of the DNA of the isolates ranged from 72.0 to 72.4 mol%. The 16S rRNA gene sequences of the isolates were very similar to each other (> or = 99.8%). The phylogenetic analysis showed that the isolates formed a cluster with species of the genus Nocardioides with Nocardioides simplex and Nocardioides nitrophenolicus as their nearest neighbors. DNA-DNA hybridization studies showed that the isolates showed a hybridization level of less than 55% to any tested species of the genus Nocardioides. Based on these data, Nocardioides aromaticivorans sp. nov. is proposed for the new DF-degrading isolates. The type strain is strain H-1 (IAM 14992, JCM 11674, DSM 15131). PMID- 15830812 TI - Quantitative assessment of sense and antisense transcripts from genes involved in antigenic variation (vsp genes) and encystation (cwp 1 gene) of Giardia lamblia clone GS/M-83-H7. AB - Antigenic variation of the intestinal protozoan parasite Giardia lamblia is caused by an exchange of the parasite's variant surface protein (VSP) coat. Many investigations on antigenic variation were performed with G. lamblia clone GS/M 83-H7 which produces surface antigen VSP H7. To generate novel information on giardial vsp gene transcription, vsp RNA levels were assessed by quantitative reverse transcription-(RT)-PCR in both axenic VSP H7-type trophozoites and subvariants obtained after negative selection of GS/M-83-H7 trophozoites by treatment with a cytotoxic, VSP H7-specific monoclonal antibody. Our investigation was not restricted to the assessment of the sense vsp transcript levels but also included an approach aimed at the detection of complementary antisense vsp transcripts within the two trophozoite populations. We found that sense vsp H7 RNA predominated in VSP H7-type trophozoites while sense RNA from only one (vsp IVg) of 8 subvariant vsp genes totally analysed predominated in subvariant-type trophozoites. Interestingly, the two trophozoite populations exhibited a similar relative distribution regarding the vsp H7 and vsp IVg antisense RNA molecules. An analogous sense versus antisense RNA pattern was also observed when the transcripts of gene cwp 1 (encoding cyst wall protein 1) were investigated. Here, both types of RNA molecules only appeared after cwp 1 had been induced through in vitro encystation of the parasite. These findings for the first time demonstrated that giardial antisense RNA production did not occur in a constitutive manner but was directly linked to complementary sense RNA production after activation of the respective gene systems. PMID- 15830813 TI - Parasitism and growth in the earthworm Lumbricus terrestris: fitness costs of the gregarine parasite Monocystis sp. AB - Parasites inflict fitness costs on their hosts, but often the exact reduction in fitness is not well understood. We investigated the influence of infection by the gregarine genus Monocystis sp. on growth and female investment (cocoon production) of its earthworm host, Lumbricus terrestris. Earthworms (n = 81) were observed in a laboratory setting for 8 months, after which parasite load was determined. The results revealed a significant negative relationship between parasite load and growth, yet no association to cocoon production was found. Although the exact nature, strength, and evolutionary consequence of reduced growth are still unclear, the results are the first indication for a clear, albeit weak effect of Monocystis on host fitness. PMID- 15830814 TI - Phylogenetic analysis of freshwater fish trypanosomes from Europe using ssu rRNA gene sequences and random amplification of polymorphic DNA. AB - The taxonomy and phylogenetic relationships of fish trypanosomes are uncertain. A collection of 22 cloned trypanosome isolates from 14 species of European freshwater fish and 1 species of African freshwater fish were examined by molecular phylogenetic analysis. The small subunit ribosomal RNA (ssu rRNA) genes of 8 clones were sequenced and compared with ssu rRNA gene sequences from a wider selection of vertebrate trypanosome isolates by phylogenetic analysis. All trypanosomes from freshwater fish fell in a single clade, subdivided into 3 groups. This clade sits within a larger, robust clade containing trypanosomes from marine fish and various amphibious vertebrates. All 22 trypanosome clones were analysed by random amplification of polymorphic DNA. The resulting dendrogram shows 3 groups, which are congruent with the groups identified in the ssu rRNA gene phylogeny. Two of the groups contain the majority of trypanosome isolates and within-group variation is slight. These groups do not separate purported trypanosome species distinguished by morphology or host origin, and thus these criteria do not appear to be reliable guides to genetic relationships among fish trypanosomes. However, we suggest that the 2 groups themselves may represent different species of fish trypanosomes. The polymorphic DNA markers we have identified will facilitate future comparisons of the biology of these 2 groups of fish trypanosomes. PMID- 15830815 TI - Proteolytic expression in Blastocrithidia culicis: influence of the endosymbiont and similarities with virulence factors of pathogenic trypanosomatids. AB - Blastocrithidia culicis is an insect trypanosomatid that presents bacterial endosymbionts. The cell-associated and secreted proteinases of the endosymbiont bearing and aposymbiotic strains were compared through the incorporation of proteinaceous substrates into sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Few qualitative changes could be detected in the proteolytic zymograms in the 2 strains studied when gelatin, casein, haemoglobin or bovine serum albumin (BSA) were tested. However, the level of proteolytic activities was significantly higher in the aposymbiotic strain. Some of the B. culicis proteins reacted in Western blots with antibodies raised against gp63, a zinc-metalloproteinase, and cruzipain, a cysteinyl-proteinase, which are virulence factors of the human pathogenic trypanosomatids, Leishmania spp. and Trypanosoma cruzi, respectively. The anti-cross-reacting determinant (CRD) antibody recognized 2 polypeptides (50 and 58 kDa) in the spent culture media and in the supernatant from glycosylphosphatidylinositol-phospholipase C (GPI-PLC) treated cells, suggesting that these proteins are GPI-anchored to the plasma membrane. In addition, the anti-gp63 reacted with the 50 kDa protein. The identification of protein homologues in trypanosomatids with distinct life-cycles may help to determine the importance of proteinases in trypanosomatids. PMID- 15830816 TI - Blood-feeding in the young adult filarial worms Litomosoides sigmodontis. AB - In this study with the filarial model Litomosoides sigmodontis, we demonstrate that the worms ingest host red blood cells at a precise moment of their life cycle, immediately after the fourth moult. The red blood cells (RBC) were identified microscopically in live worms immobilized in PBS at 4 degrees C, and their density assessed. Two hosts were used: Mongolian gerbils, where microfilaraemia is high, and susceptible BALB/c mice with lower microfilaraemia. Gerbils were studied at 12 time-points, between day 9 post-inoculation (the worms were young 4th stage larvae) and day 330 p.i. (worms were old adults). Only the very young adult filarial worms had red blood cells in their gut. Haematophagy was observed between days 25 and 56 p.i. and peaked between day 28 and day 30 p.i. in female worms. In males, haematophagy was less frequent and intense. Similar kinetics of haematophagy were found in BALB/c mice, but frequency and intensity tended to be lower. Haematophagy seems useful to optimize adult maturation. These observations suggest that haematophagy is an important step in the life-cycle of L. sigmodontis. This hitherto undescribed phenomenon might be characteristic of other filarial species including human parasites. PMID- 15830818 TI - Nematode parasites of sheep: extension of a simple model to include host variability. AB - We use results from a simulation-based model of nematode infection of sheep to refine the parameters in a simpler generic model of host-parasite population dynamics. These parameters describe the following host-parasite traits: probability of establishment of ingested larvae, mortality rate of adult parasites, and fecundity of adult female parasites. This simple model is then extended by allowing those parameters to vary amongst individual hosts. A sensitivity analysis is performed to determine which parameters have most influence on host parasite burden. The establishment parameter has the greatest effect on the peak value of parasite burden whilst the other two parameters have more effect on the duration of the burden. A comparison is made with results from the flock model after discussion of the definition of an average host. By allowing these parameters to vary simultaneously within the individual hosts we are able to reproduce the over-dispersed distribution of adult parasites frequently seen in nematode infections of sheep flocks. PMID- 15830817 TI - Characterization of proteinases in different isolates of adult Haemonchus contortus. AB - A high degree of intra- and inter-geographical variation has been demonstrated previously in the excretory/secretory proteinases released by adult Haemonchus contortus. Proteinase activity has also been associated with host-protective 'hidden' antigens isolated from the gut of adult H. contortus. If similar geographical strain variation also exists within the gut-associated proteinases, this will have important implications for the development of a globally effective vaccine. The proteinases active in integral-membrane protein extracts from 3 different strains of adult H. contortus were characterized on the basis of their pH optima and molecular size. Although enzyme activity was detected over a wide pH range, the majority of proteinase activity was detected at acidic pH. Differences in specific activity and size of enzymes were observed between the 3 different parasite strains at different pH values. A high degree of conservation in reactive peptides was observed when protein extracts were probed with antisera raised to the protective hidden gut-antigen complexes isolated from the Moredun strain of H. contortus, or to bacterially expressed subcomponents thereof. Therefore, despite the observed differences in membrane-bound proteinase profiles, the similarity of the immunogenic response against these hidden antigens may be sufficient to prove protective against different geographical isolates of H. contortus. PMID- 15830819 TI - Evidence for macrofilaricidal activity of ivermectin against female Onchocerca volvulus: further analysis of a clinical trial in the Republic of Cameroon indicating two distinct killing mechanisms. AB - During a 3-year trial of the effects of ivermectin (Mectizan) on adult worms and microfilariae of Onchocerca volvulus in the Republic of Cameroon, comparison was made between the percentages of calcified and uncalcified moribund (M) and dead (D) adult female worms dying following (a) the standard dose (150 microg/kg) given annually; (b) high doses (400, then 800 microg/kg) given annually; and (c and d) these same doses given at 3-monthly intervals. In the killing of adult female O. volvulus worms, the relative roles of (a) natural causes; (b) a presumed, direct, anthelminthic, macrofilaricidal action of ivermectin; and (c) a potentially fatal pleomorphic ovarian neoplasm (PN), of which the incidence is increased by ivermectin treatment, are herein further investigated and discussed. It is concluded that ivermectin per se has a considerable direct macrofilaricidal action against female worms and that this lethal effect is supplemented by the drug's ability in some worms to increase the incidence, and the spread throughout the body of the worm, of the potentially fatal PN ovarian tumour. In moribund and dead ivermectin-treated female worms that were heavily invaded by PN, it is probable that the neoplasm was chiefly responsible for their death, but the additional direct anthelminthic action of the drug, which by itself has been responsible for the death of many other female worms, cannot be excluded as having played a supplementary lethal role. Similar problems as to the exact means by which adult female worms are killed may arise now that ivermectin is used in Africa for the mass treatment of lymphatic filariasis; or if and when the macrofilaricidal actions on O. volvulus of other drugs, which are closely related to ivermectin, come to be investigated. PMID- 15830820 TI - Cystic echinococcosis: analysis of the serological profile related to the risk factors in individuals without ultrasound liver changes living in an endemic area of Tacuarembo, Uruguay. AB - Cystic echinococcosis (CE) is a high prevalence zoonosis among the rural population of Tacuarembo (Uruguay). The correlation between serological data and the incidence of risk factors was studied in a survey carried out in 1998 among rural communities where 480 individuals were examined by means of abdominal sonography (local prevalence = 0.8%). Serum samples (305) were analysed by ELISA to determine specific IgG against crude antigens from Echinococcus granulosus. A total of 27 individuals exhibiting no detectable changes in abdominal sonographic examination were found to be seropositive ('ultrasound normal group'). Of these individuals 9 were seroreactive against purified antigen B. A significant degree of correlation was found between seroreactivity and the incidence of some risk factors (CE antecedent in the family, P < 0.005 and use of rural water, P < 0.0001) among this group. Follow-up of individuals of the 'ultrasound normal group' was carried out after 2 years to evaluate the implications of this serological reactivity. No predictive value for cyst development was assessed with complementary image study; in contrast transient antibodies were observed with both crude and purified antigen as approximately 60% of individuals became negative when re-sampled. PMID- 15830821 TI - Morphological and molecular characterization of tetraphyllidean merocercoids (Platyhelminthes: Cestoda) of striped dolphins (Stenella coeruleoalba) from the Western Mediterranean. AB - Two types of tetraphyllidean merocercoids, Phyllobothrium delphini and Monorygma grimaldii, are well known from most cetaceans world-wide. The role of cetaceans in the life-cycle of these merocercoids is unclear because their specific identity is as yet unknown. The problem is compounded by poor descriptions of both merocercoids. We used light and scanning electron microscopy, and histological techniques to provide a thorough description of merocercoids collected from 11 striped dolphins, Stenella coeruleoalba, from the Spanish Mediterranean. We also described, for the first time, specimens of P. delphini with immature proglottides. Our merocercoids were morphologically similar to those described previously, except in the structure of the apical organ. Intra- and inter-sample variability in the morphology of the apical organ suggested that it degenerates during larval development. A subsample of 16 specimens of P. delphini and M. grimaldii was characterized for the D2 variable region of the large subunit ribosomal RNA gene (LSU) and compared with published tetraphyllidean cestode LSU sequences. P. delphini showed 2 unique signatures that differed from one another by a single base, whereas all sequences of M. grimaldii were identical. This suggests that each type may represent a single species, contrary to previous speculations based on morphological data. All merocercoid specimens formed a clade together with Clistobothrium montaukensis. Based on the low degree of divergence, all specimens of this clade are predicted to be congeneric. PMID- 15830822 TI - Expression of depressive symptoms in a nonclinical Brazilian adolescent sample. AB - OBJECTIVE: This study aimed to detect the prevalence of depressive symptomatology and its expression in a nonclinical Brazilian adolescent student sample. METHOD: A sample of students from private and public schools (n = 1555, aged 13 to 17 years) answered the Beck Depression Inventory (BDI). We performed factor analysis of the BDI as an indicator of the expression of depressive symptomatology. The following cut-off scores defined nonclinical subgroups: "nondepressed," BDI < 15; "dysphoria," BDI 16 to 20; and "depressed," BDI > 20. We used discriminant analysis to test whether these subgroups could be separated by the depression specific and nonspecific items. RESULTS: The point prevalence of depression was 7.6%, according to the BDI cut-off of 20. Girls had higher scores than boys in several items. Scores increased with age. Students from public schools had higher scores than did private school students. Factor analysis showed 2 common factors for the total sample and for each sex: the cognitive affective dimension and the somatic nonspecific dimension. In the adolescents showing clinical depression, items related to self-depreciation, sense of failure, guilty feelings, self dislike, suicidal wishes, and distortion of body image were common components of BDI factors. Discriminant analysis showed that the BDI highly discriminates depressive symptomatology in adolescent students and also measures specific aspects of depression. CONCLUSIONS: The BDI is useful as a measure of specific aspects of depression in nonclinical adolescent samples; it was able to detect depression in approximately 7% of the surveyed population. The expression of depressive symptoms in a Brazilian adolescent population is compatible with international studies in this age group. Detecting depressive symptoms in a school population is a critical preventive strategy; to avoid damage to the learning process, it should be followed with further referral to treatment when needed. PMID- 15830823 TI - Posttraumatic stress disorder and general psychopathology in children and adolescents following a wildfire disaster. AB - OBJECTIVE: To report on the use of the Post Traumatic Stress Disorder Reaction Index (PTSD-RI) and the Strengths and Difficulties Questionnaire (SDQ) in identifying children and adolescents who may require psychological interventions following exposure to a wildfire disaster. METHOD: Six months after a wildfire disaster, we conducted a school-based program to screen for wildfire-related events, such as exposure to and perception of threat, posttraumatic stress disorder (PTSD), and general psychopathology. RESULTS: The screening battery was completed by 222 children (mean age 12.5 years, SD 2.48; range 8 to 18 years). Severe or very severe PTSD was reported by 9.0% of students, while 22.6% scored in the abnormal range on the Emotional Symptoms subscale of the SDQ. Younger children and individuals with greater exposure to and perception of threat experienced higher levels of PTSD and general psychopathology. Female students reported a greater perception of threat but did not report higher levels of PTSD or other symptoms. CONCLUSIONS: Screening was well received by students, parents, and staff and proved feasible in the postdisaster environment. The PTSD-RI and SDQ demonstrated different individual risk associations and functioned as complementary measures within the screening battery. The identification of children at greatest risk of mental health morbidity enabled service providers to selectively target limited mental health resources. PMID- 15830824 TI - Individual change in methylphenidate use in a national sample of children aged 2 to 11 years. AB - OBJECTIVES: To determine methylphenidate use in children aged 2 to 13 years. To provide age- and sex-specific estimates of methylphenidate initiation and cessation during a 2-year period. METHOD: Data from 2 cycles of a Canadian household survey yielded a sample of over 10 000 children aged 2 to 11 years at Cycle 1 who continued to participate at Cycle 2. We used logit modelling to estimate Cycle 2 methylphenidate use, methylphenidate use over a 2-year period, and methylphenidate initiation and cessation from Cycles 1 to 2. RESULTS: In 1996 and 1997, methylphenidate use ranged from 0.32% to 6.31% among children aged 4 to 13 years. School-aged boys were more likely than girls to use methylphenidate. Odds were greater for boys aged 6 to 7 years than for boys aged 4 to 5 years; they were also greater for boys aged 10 to 11 years than for boys aged 12 to 13 years. Almost 1% of children used methylphenidate at both data cycles. Odds of Cycle 2 methylphenidate use were 135 times greater for children using methylphenidate at Cycle 1, compared with nonusers. Methylphenidate initiation ranged from 0.20% to 3.34%, and school-aged boys had higher initiation rates than girls. Cessation rates ranged from 18% to78%, and there were no statistically significant differences by age and sex. CONCLUSIONS: Methylphenidate prevalence findings are consistent with past studies. We found an age-by-sex interaction on methylphenidate use. We also found both continuity and discontinuity in methylphenidate use. PMID- 15830825 TI - [Methadone and sleep apnea syndrome]. AB - BACKGROUND: Sleep apnea syndrome occurs when, during sleep, breathing stops for 10 seconds or longer, with an index of 5 times or more an hour. It is clinically characterized by loud snoring at night, continuous or interrupted by pauses followed by loud breathing. Sleep is fitful, broken by arousals, and yields little rest. There is daytime excessive sleepiness with repeated involuntary falling asleep, often unknown by the subject. CLINICAL DESCRIPTION: In this article, we describe an observation of central sleep apnea syndrome in a female patient receiving an opiate replacement therapy. METHOD: An analysis of the before and after methadone withdrawal polysomnograhic tracing was done for this patient. RESULTS: This diagnosis etiology and physiopathology are critically approached. Clinicians should be careful in treating induced sleep disorders in such patients. CONCLUSION: Prescribing benzodiazepines during an opiate withdrawal of the methadone type is not recommended when central apnea occurs. PMID- 15830826 TI - An introduction to economic evaluation: what's in a name? AB - OBJECTIVE: This paper describes the main types of economic evaluation techniques. METHOD: To examine the strengths and limitations of different types of economic evaluations, we used a hypothetical example to review the reasoning underlying each method and to illustrate when it is appropriate to use each method. RESULTS: The choice of economic evaluation method reflects a decision about what should represent "success" and how success should be valued. Measures of benefit and cost must be considered systematically and simultaneously. Claiming that a new treatment is cost-effective requires making a value judgment based on the personal beliefs of the claimant. Even when cost and effect data are objective, a verdict of cost-effective is subjective. The conclusions of an economic study can change significantly, depending on which patient outcome is used to measure success. CONCLUSIONS: Clinicians must be sure that important patient outcomes are not excluded from economic evaluations. Economic evaluation is a process designed to produce an estimate rather than a decision. New treatment can be more costly and still be cost-effective (if the extra benefit is valued more than the extra cost to produce it). However, since economic evaluation does not explicitly consider a decision maker's available budget, a new treatment can be deemed cost effective but too expensive to approve. PMID- 15830827 TI - Using metaanalysis to evaluate evidence: practical tips and traps. AB - Although practising evidence-based medicine is the goal of most physicians, it can be a real challenge to sift through the vast body of data to determine the best strategies. Most clinical guidelines regard replicated randomized controlled trials (RCTs), metaanalyses, and systematic reviews as the highest level of evidence to support treatment recommendations. High-quality metaanalyses can overcome many of the drawbacks of individual RCTs and qualitative reviews. They can reduce bias, provide adequate power to demonstrate real differences in outcomes, and resolve the results of inconsistent studies. This paper focuses on basic principles and terms used in metaanalysis, so that clinicians can appropriately evaluate and use their results to guide treatment decisions. PMID- 15830828 TI - Experimental affective symptoms in panic disorder patients. AB - OBJECTIVE: To date, carbon dioxide (CO2) challenge tests in panic disorder (PD) patients have focused on anxiety as the sole outcome measure. This study assesses a broader range of symptoms in patients with PD. METHOD: We administered a gas mixture of 35% CO2 and 65% oxygen (O2) to 25 patients with PD. Nine patients met the criteria for a comorbid major depressive disorder (MDD), and 16 did not. We assessed not only subjects' symptoms of anxiety but also their symptoms of depression and aggression. RESULTS: Baseline ratings did not differ across the 2 subgroups. Postchallenge ratings were higher for PD and MDD patients on all the assessed affective symptoms, except for specific panic symptoms. CONCLUSION: These findings suggest that, in addition to anxiety, CO2 challenge induces depressive and aggressive symptoms, specifically in PD patients with comorbid depression. PMID- 15830829 TI - Hemorrhages during escitalopram-venlafaxine-mirtazapine combination treatment of depression. PMID- 15830830 TI - Re: Lorazepam-induced prolongation of the QT interval in a patient with schizoaffective disorder and complete AV block. PMID- 15830831 TI - Lithium-associated anencephaly. PMID- 15830832 TI - Aripiprazole-induced seizure. PMID- 15830833 TI - Prevalence of bipolar disorder and major depression among patients seen in primary and secondary care in Finland. PMID- 15830834 TI - The need for more community nursing for adults with intellectual disabilities and mental health problems. PMID- 15830835 TI - Improvement in tardive dyskinesia with aripiprazole use. PMID- 15830836 TI - Antidepressant action of St. John's Wort, Hypericum perforatum: a test of the circadian hypotheses. AB - Extracts of the plant St. John's Wort, Hypericum perforatum, are effective for treatment of mild depression. It has been hypothesised that H. perforatum may be acting on the circadian timing system either directly or via a photosensitising action to produce changes in mood. Two experiments were conducted to test these hypotheses. Under constant dark (Experiment 1) or low constant light (Experiment 2) rats were permitted to free-run. Rats were then treated with a 'high' (616mg/kg/day; n = 8 per experiment) or 'low' (308 mg/kg/day; n = 8 per experiment) dose of H. perforatum or a control solution (n = 8 per experiment) in drinking water, and circadian locomotor rhythms examined for alterations of period. A minor shortening of mean period (2.4 min) was observed on cessation of H. perforatum treatment in the low-dose group in Experiment 2, and was considered to be a measurement artifact and of no clinical value. Otherwise, no significant differences in free-running period between treatment groups were observed in either study, indicating that H. perforatum has no direct or photosensitising effect on the mammalian circadian system. These results suggest that the antidepressant action of H. perforatum is not mediated by a circadian mechanism. PMID- 15830837 TI - Magnesium as NMDA receptor blocker in the traditional Chinese medicine Danshen. AB - Aqueous extracts of the traditional Chinese medicine Danshen, the dried roots of Salvia miltiorrhiza Bunge (Labiatae), blocked N-methyl-D-aspartate (NMDA) evoked currents in cerebrocortical neurons in vitro. The block of the NMDA-evoked currents was voltage dependent and showed the negative slope conductance reminiscent of the effect of Mg2+ ions. Atomic absorption spectrophotometry (AAS) revealed that aqueous Danshen extracts contained approximately 9mM magnesium. Fractionation of the extracts by high performance liquid chromatography followed by patch clamp recording and AAS indicated that magnesium ions were present in two distinct fractions. One fraction contained approximately 5 mM magnesium and blocked NMDA-induced currents indicating that it contained mostly free Mg2+ ions, while a second fraction did not possess NMDA antagonist activity despite the presence of approximately 4 mM magnesium suggesting that Mg2+ in this fraction was mostly chelated. Following removal of the free Mg2+ by ion exchange chromatography, the previously observed block of the NMDA-induced currents was abolished. These data demonstrate that Danshen contains both free and chelated Mg2+. Free Mg2+ ions account for the NMDA antagonist activity of Danshen in vitro. PMID- 15830838 TI - Cimicifuga racemosa extract BNO 1055 inhibits proliferation of the human prostate cancer cell line LNCaP. AB - Extracts from black cohosh (Cimicifuga racemosa, CR) exert an anti-proliferative action in human breast cancer cell cultures, which has been attributed to an anti estrogenic effect. However, CR constituents do not bind to either of the known estrogen receptors. Thus, the anti-tumor effect of CR me be mediated by mechanisms not involving these receptors. Polycyclic aromatic hydrocarbons are toxic environmental pollutants, which indirectly act as anti-estrogens by activating the aryl hydrocarbon receptor (AhR). The AhR is widely expressed in mammalian tissues and tumors. A recent screening study demonstrated activation of the AhR by a variety of herbal extracts, among others, CR. Since activation of the AhR causes inhibition of growth of prostate cancer cells, we addressed the question, whether CR may not only inhibit growth of breast cancer--but also of prostate cancer cells. In the AhR ligand assay, the CR extract BNO 1055 reduced tracer binding to 71% of the control demonstrating interaction of constituents of this extract with the receptor. Under basal as well as under estradiol- and dihydrotestosterone stimulated conditions, the CR extract dose dependently inhibited proliferation of LNCaP cells. A significant reduction of cell growth was observed at a concentration as low as 50 ng/ml. Thus, it is demonstrated for the first time that CR compounds potently inhibit the growth of human prostate cancer cells in vitro. This anti-proliferative effect may be mediated via the AhR. PMID- 15830839 TI - Fangchinoline inhibited the antinociceptive effect of morphine in mice. AB - Fangchinoline (FAN), a non-specific calcium antagonist, is a major alkaloidal component of the creeper Stephania tetrandra S. Moore (or fenfangji). It has been shown to possess antagonistic activity on morphine-induced antinociception in mice. This study was undertaken to assess the antagonistic mechanism. The results demonstrated that FAN (IP) attenuated morphine (SC)-induced antinociception in a dose-dependent manner with significant effect at doses of 30 and 60mg/kg body wt. (IP) in the tail-flick test but not the tail-pinch tests, carried out in mice. This antagonism was abolished by pretreatment with a serotonin precursor, 5 hydroxytryptophan (5-HTP, IP), but not by pretreatment with a noradrenaline precursor, L-dihydroxyphenylalanine (L-DOPA, IP) in the tail-flick test. On the other hand, the development of morphine-induced analgesic tolerance was not prevented by FAN. These results suggest that the serotonergic pathway may be involved in the antagonism of morphine-induced antinociception by FAN and, in agreement with other reports, also indicates the possible dissociation of the morphine analgesic effect from its tolerance-development mechanism. PMID- 15830840 TI - Osteoblastic proliferative activity of Epimedium brevicornum Maxim. AB - The effect of the extracts of Epimedium brevicornum Maxim. was investigated on proliferative activity in vitro. The osteoblast-like UMR106 cells was employed as an osteoblast model. The EtOH extract and the n-butanol fraction from the crude extract were found to show proliferation stimulating activity. Three flavonoid compounds (icariin, epimedin B and epimedin C) were isolated from this fraction by activity-guided assay, and the effects on cell proliferation were studied. Icariin produced the most significant promoting effect on the proliferation in osteoblast-like UMR106 cells. The results suggested that E. brevicornum Maxim. extracts might have potential activity against osteoporosis, and flavonoids such as icariin might be the active constituents stimulating osteoblasts. PMID- 15830841 TI - In vitro induction of endothelial cell fibrinolytic alterations by Nigella sativa. AB - The effect of Nigella sativa (NS) L. oil (blackseed oil) on the fibrinolytic system of the human umbilical vein (HUV) and human uterine arterial (HUA) endothelial cells (ECs) in culture was studied. Both of them showed a concentration-dependent increase in tissue-type plasminogen activator (t-PA). A maximum effect was achieved with 50 microg oil/ml conditioned medium (CM) (1.3+/ 0.15ng/10(4) cells/24h vs. control 0.7+/-0.06ng/10(4) cells/24h, and 0.38+/ 0.04ng/10(4) cells/24h vs. control 0.24+/-0.02ng/10(4) cells/24h, for HUVEC and HUA-EC, respectively). At 100 microg/ml, there was a significant change in the amount of t-PA antigen produced by either HUVEC or HUA-EC (1.0+/-0.1 ng/10(4) cells/24 h or 0.28+/-0.02 ng/10(4) cells/24 h) as compared to control CM from cells grown under control conditions, but still less than that recorded at 50 microg oil/ml. Plasminogen activator inhibitor-type 1 increased the CM significantly and concentration-dependently in both cells. For HUVEC, the maximum effect was achieved at a concentration of 100 microg/ml (257.7+/-8.0 ng/10(4) cells/24 h vs. control 72.7+/-3.8 ng/10(4) cells/24 h). HUA-EC showed the maximum effect at a concentration of 100 microg/ml (171.6+/-4.4 ng/10(4) cells/24 h vs, control 53.8+/-3.7 ng/10(4) cells/24 h). This study suggests a role for NS oil in modulating the balance of fibrinolysis/thrombus formation by modulating the fibrinolytic potential of endothelial cells. PMID- 15830842 TI - Antibacterial activity of alpha-mangostin against vancomycin resistant Enterococci (VRE) and synergism with antibiotics. AB - alpha-Mangostin, isolated from the stem bark of Garcinia mangostana L., was found to be active against vancomycin resistant Enterococci (VRE) and methicillin resistant Staphylococcus aureus (MRSA), with MIC values of 6.25 and 6.25 to 12.5 microg/ml, respectively. Our studies showed synergism between alpha-mangostin and gentamicin (GM) against VRE, and alpha-mangostin and vancomycin hydrochloride (VCM) against MRSA. Further studies showed partial synergism between alpha mangostin and commercially available antibiotics such as ampicillin and minocycline. These findings suggested that alpha-mangostin alone or in combination with GM against VRE and in combination with VCM against MRSA might be useful in controlling VRE and MRSA infections. PMID- 15830843 TI - Radioprotection by mangiferin in DBAxC57BL mice: a preliminary study. AB - The radioprotective effects of various concentrations (0, 0.25, 0.5, 1, 2, 5, 10, 17.5, 25, 50, 75 and 100 mg/kg b.wt.) of mangiferin (MGN) was studied in the DBAxC57BL mice whole body exposed to 10 Gy of gamma-irradiation. Treatment of mice with different doses of MGN, one hour before irradiation reduced the symptoms of radiation sickness and delayed the onset of mortality when compared with the non-drug treated irradiated controls. The radioprotective action of MGN increased in a dose dependent manner up to 2mg/kg and declined thereafter. The highest radioprotective effect was observed at 2mg/kg MGN, where greatest number of animals survived against the radiation-induced mortality. The administration of 0.5, 1, 2, 5, 10 and 17.5 mg/kg MGN reduced the radiation-induced gastrointestinal death as evident by a greater number of survivors up to 10 days in this group when compared with the DDW + 10 Gy irradiation group. A similar effect of MGN was observed for the radiation-induced bone marrow deaths also. Our study demonstrates that mangiferin, a gluosylxanthone, present in the Mangifera indica protected mice against the radiation-induced sickness and mortality and the optimum protective dose of 2mg/kg was 1/200 of LD50 dose (400 mg/kg) of MGN. The administration of 400 mg/kg MGN induced 50% mortality, therefore LD50 of the drug was considered to be 400 mg/kg. PMID- 15830844 TI - Potent suppressive effect of green tea polyphenols on tobacco-induced mutagenicity. AB - Antimutagenic activity of green tea (Camellia sinensis) was studied using Salmonella typhimurium strains (TA 102) (Ames test). Aqueous tobacco extract was found to be mutagenic to S. typhimurium TA 102 at concentration of 50 mg/plate. Green tea polyphenols was found to inhibit the mutagenicity of tobacco in a concentration-dependent manner. Concentrations needed for 50% inhibition of mutagen-induced revertant formation was found to be 5 mg/plate. Green tea polyphenols was also found to inhibit the urinary mutagenicity in rats induced by tobacco extract. Moreover green tea polyphenols were found to inhibit in vitro nitrosation reaction produced by reaction sodium nitrite and methyl urea and further inhibition of mutagenicity indicating that green tea has dual action to bring out a reduction in the mutagenic and carcinogenic potential of tobacco. PMID- 15830845 TI - Antibacterial activity of Turkish propolis and its qualitative and quantitative chemical composition. AB - The antibacterial activity of propolis from different regions of Turkey was studied, accompanied by TLC and GC-MS analyses of its chemical composition and spectrophotometric quantification of the most important active principles. All six samples were active against the bacterial test strains used; however, samples 1 (Yozgat), 2 (Izmir) and 3 (Kayseri) were more active than samples 4 (Adana), 5 (Erzurum) and 6 (Artvin). By TLC comparison all samples were found to contain poplar taxonomic markers but in samples 4 (Adana), 5 (Erzurum) and 6 (Artvin), different substances were observed, which were not present in P. nigra L. bud exudate. The typical poplar samples 1 (Yozgat), 2 (Izmir) and 3 (Kayseri) displayed very similar phenolic and flavonoid content. Samples 4 (Adana), 5 (Erzurum) and 6 (Artvin) were characterized by low phenolic and very low flavonoid concentrations. Qualitative analysis by GC-MS revealed that sample 4 (Adana) contained diterpenic acids and high percent of cinnamyl cinnamate, sample 5 (Erzurum)-significant amounts of hydroxy fatty acids and triterpenic alcohoLs, and sample 6 (Artvin)-phenolic glycerides, characteristic for the bud exudate of Populus euphratica Oliv. The results confirm the importance of phenolics for propolis antibacterial activity, and the significance of P. nigra L. as a propolis source, which provides the hive with the best defense against microorganisms. PMID- 15830846 TI - Antibacterial efficacy of Withania somnifera (ashwagandha) an indigenous medicinal plant against experimental murine salmonellosis. AB - In the present study, we evaluated the antibacterial activity of ashwagandha [Withania somnifera L. Dunal (Solanaceae; root and leaves)], an Indian traditional medicinal plant against pathogenic bacteria. Both aqueous as well as alcoholic extracts of the plant (root as well as leaves) were found to possess strong antibacterial activity against a range of bacteria, as revealed by in vitro Agar Well Diffusion Method. The methanolic extract was further subfractionated using various solvents and the butanolic sub-fraction was found to possess maximum inhibitory activity against a spectrum of bacteria including Salmonella typhimurium. Moreover, in contrast to the synthetic antibiotic (viz. chloramphenicol), these extracts did not induce lysis on incubation with human erythrocytes, advocating their safety to the living cells. Finally, the antibacterial efficacy of the extracts isolated from plant (both root and leaves) was determined against experimental salmonellosis in Balb/C mice. Oral administration of the aqueous extracts successfully obliterated salmonella infection in Balb/C mice as revealed by increased survival rate as well as less bacterial load in various vital organs of the treated animals. PMID- 15830847 TI - Antifungal activity of some Brazilian Hypericum species. AB - Crude methanolic extracts and fractions from the aerial parts of seven species of Hypericum (H. caprifoliatum Cham. and Schltdl., H. carinatum Griseb., H. connatum Lam., H. ternum A. St.-Hil., H. myrianthum Cham. and Schltdl., H. piriai Arechav. and H. polyanthemum Klotzsch ex Reichardt) growing in southern Brazil were analyzed for their in vitro antifungal activity against a panel of standardized and clinical opportunistic pathogenic yeasts and filamentous fungi, including dermatophytes, by the agar dilution method. Chloroform and hexane extracts of H. ternum showed the greatest activity among extracts tested. PMID- 15830848 TI - Fumigant toxicity of the essential oils of some African plants against Anopheles gambiae sensu stricto. AB - The essential oils from 15 species of African plants selected by ethnobotanical considerations and field inspection (odour and presence of insects) were screened for fumigant toxicity to Anopheles gambiae s.s. in the laboratory. Essential oils from 6 species showed varying levels of toxicity, with Conyza newii (Compositae) and Plectranthus marruboides (Labiateae) being the most potent. Fifty compounds representing approximately 74% of the essential oil of C. newii were identified by GC-MS and GC-coinjection (for available standards). The major and some of the minor constituents of the two oils were assayed at different doses. Two compounds, from C. newii, perillaldehyde and perillyl alcohol, exhibited higher fumigant toxicity (LD50 = 1.05 x 10(-4) and 2.52 x 10(-4) mg cm(-3), respectively) than the parent oil (2.0 x 10(-3) mg cm(-3)). GC-MS analysis of the essential oil of P. marruboides gave results similar to that previously reported. Interestingly, none of its components were active, suggesting that the insecticidal activity of the oil results from either some of the minor components or as a blend effect of some of the major constituents. PMID- 15830849 TI - Adverse reaction to an adrenergic herbal extract (Citrus aurantium). AB - We report the case of a 52 year old woman that had an adverse reaction after taking a dry herbal extract of an unripe fruit of Citrus aurantium L. var. amara, as dietary supplement for weight loosing. The fruit is also known as zhi shi (in traditional Chinese Medicine) or bitter orange in other parts of the world. PMID- 15830851 TI - [A sampling survey on the distribution of Oncomelania snails in Zhejiang Province]. AB - OBJECTIVE: To find out current distribution of Oncomelania snails and Schistosoma infection in snails in Zhejiang Province, so as to improve the project of schistosomiasis control. METHODS: Investigation spots were selected by stratified cluster sampling method, 100 villages of 34 counties were selected from 7106 villages of 55 counties as survey spots. Synchronously systematic and environmental samplings were used for the survey. Snails were dissected to determine the infection status and spots with sham snails were set to assess the quality of the survey. RESULTS: The result showed that snails were found in 223 strips, 1572 frames and 73,300 m2 area in 32 villages of 21 counties. Snails were found in an area covering 72,640 m2 in 29 villages of 18 counties in hilly region, which accounted for 99.1% of total snail habitats. The significantly larger area with snails was revealed in hilly region than that in water network region (t = 3.04, P < 0.01). 1st, 2nd strata occupied 71,680 m2 snail area, which accounted for 97.8% of total snail area, much higher than that in the 3rd stratum (t = 3.71 , P < 0.01). Snail habitats in the irrigation ditches and canals accounted for 62.6% of total snail area, significantly higher than that of farmland (t = 3.02, P < 0.01). No snails were found infected among 7892 living snails by dissection. The theory value of snail area in Zhejiang Province was 1,361,940 m2. CONCLUSION: It is estimated that the snail-ridden area is about 1,500,000-2,000,000 m2 in the Province, mostly found in the villages of hilly region with a record of snail habitats during the recent 4 years and the primary environment of the snail areas were ditches and canals. No infected snails were found from the survey. PMID- 15830850 TI - [Distribution of inducible nitric oxide synthase in different stages of Schistosoma japonicum]. AB - OBJECTIVE: To study the presence of iNOS in Schistosoma japonicum and demonstrate its distribution in different stages of this schistosome. METHODS: Cryostat sections for adult worms and paraffin sections for eggs in the liver of infected mouse, sporocysts and cercariae in snails were prepared, immunofluorescent test was performed to detect the presence of iNOS in adult worms, sporocysts, cercariae and miracidium, the distribution of this enzyme was observed in different stages of Schistosoma japonicum. Western blotting was used to further demonstrate the immunoreactivity to iNOS in adult worms. RESULTS: The results of immunofluorescent test showed that specific yellow-green fluorescence was mainly among subtugment of adult worms. Positive staining was also distributed on the surface of miracidium and its glands. For both sporocysts and cercariae, the majority of fluorescence was on their surface. Anti-iNOS antibody could recognize an apparent specific band in Western blotting of adult worm proteins, with a of Mr 210,000. CONCLUSION: There is an expression of iNOS-like enzyme in Schistosoma japonicum. PMID- 15830852 TI - In vitro effect of cyclosporine A on juvenile Schistosoma mansoni labeled by AF18 (5-N-octadecanoyl aminofluorescein). AB - OBJECTIVE: To explore the in vitro effect of cyclosporine A on the tegument of juvenile Schistosoma mansoni labeled by AF18 and investigate the effect of cyclosporine A on schistosomula surface membrane fluidity. METHODS: Preparation of transformed schistosomula, adding cyclosporine A into tubes containing schistosomula and labeling of transformed schistosomula with AF18, then observe schistosomula under fluorescence microscope. RESULTS: Schistosomula of different groups labeled by AF18 were damaged by cyclosporine A in vitro. CONCLUSION: Cyclosporine A increases the uptake of AF18 by schistosomula in vitro which is dose-dependent, and decreases the parasite surface membrane fluidity. PMID- 15830853 TI - [Case report: skin myiasis]. PMID- 15830854 TI - [The survey of Cryptosporidium infection among young children in kindergartens in Anhui Province]. AB - OBJECTIVE: To study the cryptosporidiosis infection among young children in 29 kindergartens in the province, and to explore the best way to diagnose the infection. METHODS: Stool specimens of 1204 children were collected, oocysts of Cryptosporidium were identified with auramine O-modified staining, acid-fast staining, safranine T and methylene blue staining, and auramine O-modified acid fast staining. RESULTS: The detection rate of Cryptosporidium in four stainings were 2.46%, 1.50%, 1.98% and 3.46% respectively, and the rate was significantly higher by auramine O-modified acid-fast staining than other stainings (P < 0.01). The infection rate was significantly lower in urban children (2.14%, 15/684) than the rural ones (5.19%, 27/520). Boys and girls showed similar detection rate (1.99%, 24/1204 vs 1.50%, 18/1204). Cryptosporidium infection was usually subclinical, the major clinical features of cryptosporidium included benign diarrhea, mild abdominal pain and nausea. CONCLUSION: The Cryptosporidium infection was relatively common in kindergartens in Anhui and higher infection rate was found in rural children. As majority of the Cryptosporidium infections are subclinical, the confirmation of diagnosis is important though difficult. PMID- 15830855 TI - [Investigation and control of malaria outbreaks in 2001-2003 in Shangshui county]. PMID- 15830856 TI - [Immunoprotection induced by recombinant plasmids pVAX1-Mzp5-7 of Eimeria tenella]. AB - OBJECTIVE: To observe the immunoprotection of recombinant plasmids of lambda Mzp5 7 to chicken challenged with Eimeria tenella (E. t) oocysts. METHODS: All the chickens were immunized with the recombinant plasmids by different inoculation pathways, three times on d7, d14 and d21, and challenged with E. t oocysts. The response of specific humoral and cell immunity were detected by the immunological methods such as the specific antibody response, the ratio of CD4+/CD8+ T cells. RESULTS: The results showed that the recombinant plasmids can induce immune response. The specific immune responses were strengthened with the increase of immunization times. The difference of the immunity indexes among experiment groups was not significant, while the ratio of CD4+/CD8+ T cells and antibody titer were significantly higher in experiment groups than those in the control groups. The number of oocysts shed in experiment groups decreased and the shedding duration was shorter significantly. The body weight of the chicken in experiment groups increased significantly and the lesion in cecum was slighter than that of control groups. CONCLUSION: The recombinant plasmids of lambda Mzp5 7 show an immunoprotection to chicken challenged with E. t oocysts. PMID- 15830857 TI - [The effect of sandfly control on the transmission of visceral leishmaniasis]. AB - OBJECTIVE: To observe the effect of Phlebotomus chinensis control including insecticide residual spraying in the habitats and bathing for dogs on the transmission of visceral leishmaniasis in Nanping of Sichuan Province. METHODS: Alpha-methrin with a dosage of 50 mg/m2 was sprayed in the wild caves and 2.5% deltamethrin wettable power at a concentration of 250 mg/L was applied for dog bathing in the villages. The density of sandflies in the natural caves was examined and data on the incidence of visceral leishmaniasis were collected from epidemiological survey following the sandfly control measures. RESULTS: The density of sandflies has been considerably reduced after the chemical spraying in caves, the important habitats of the sandflies. By both the cave spraying and bathing for dogs since 1993, the reported human cases of leishmaniasis also decreased. CONCLUSION: Measures for sandfly control including insecticide spraying in the habitats and bathing for dogs with insecticide solution can significantly reduce the sandfly density, and can contain the transmission of visceral leishmaniasis in the endemic area. PMID- 15830858 TI - [Influence of interferon gamma treatment on expression of TGF-beta1 and its receptors in liver fibrosis of mice with schistosomiasis japonica]. AB - OBJECTIVE: To investigate the effect of interferon-gamma (IFN-gamma) on the expression of TGF-beta1 and its two membrane receptors--TGF-beta receptor I (TbetaRI), TGF-beta receptor II (TbetaRII), and observe the expression of TGF beta1, TbetaRI and TbetaRII during the development of liver fibrosis in BALB/c mice infected by Schistosoma japonicum. METHODS: BALB/c mice, aged 6-8 weeks, were infected with cercariae of S. japonicum. The infected mice were divided randomly into three groups 16 week after infection: model group, praziquantel group and praziquantel combined with IFN-gamma group. Liver specimen were obtained at 8, 12, 16 week and at the end of treatment. Pathological examination, immunohistochemistry and RT-PCR were used to evaluate the pathological change, the expression of TGF-beta1, TbetaRI and TbetaRII and the mRNA level respectively. RESULTS: The expression of TGF-beta1, TbetaRI, and TbetaRII can be detected in infected mice, while the expression around egg granulomas enhanced along with the progress of the disease. With the therapy of IFN-gamma, the reduction of egg granulomas, and of the expression of TGF-beta1, TbetaRI and TbetaRII was observed. From the transcription level, it was found that TGF-beta1 mRNA increased at 12 week and peaked at model group, then decreased to the normal level after treatment with IFN-gamma combined with praziquantel. The level of TbetaRII mRNA reduced at 8 and 16 week and returned to normal at the end of treatment. More interestingly, TbetaRI mRNA remained at the normal level on the whole course both in the development of fibrogenesis and the period of treatment. CONCLUSION: The up regulation of TGF-beta1 and down regulation of TbetaRII mRNA may induce liver fibrogenesis and IFN-gamma might suppress TGF-beta1 to reverse fibrosis. The mechanism of the suppression is mediated by down regulation of expression of its two receptors at protein level but not by influencing the mRNA expression. PMID- 15830859 TI - Effect of sodium citrate based anticoagulants on the growth activity of malaria parasites. AB - OBJECTIVE: To study the effect of anticoagulants based on sodium citrate on the growth activity of malaria parasites. METHODS: The parasites were treated with 3 anticoagulants (ACD, CD and SC), respectively, and the parasitemia was determined to measure the effect of the anticoagulants on the growth of the parasites. Unsynchronized Plasmodium falciparum was treated with the anticoagulants at different concentrations for 3 h at 37 degrees C. Treated erythrocytes were mixed with normal parasites or treated parasites with normal erythrocytes, which was followed by parasitemia determination of the two cultures to determine the cell target of the anticoagulants. Stage-synchronized parasites (ring, trophozoite and schizont) were treated as above to investigate the stage target. P. berghei was also treated with anticoagulants and inoculated in mice to detect the effect of anticoagulants on the animal malaria parasite by counting the parasitemia. RESULTS: All 3 anticoagulants inhibited falciparum parasite growth and ACD had the strongest potency. The treatments of the erythrocyte and the parasite with anticoagulants respectively showed that the anticoagulants targeted the parasites rather than normal erythrocytes. Stage-synchronized parasite treatment suggested anticoagulants primarily inhibited schizonts. The effect of anticoagulants on P. berghei was similar to that on P. falciparum. CONCLUSION: ACD showed the most significant inhibitive effect on the growth of malaria parasites and SC was the best anticoagulant based on sodium citrate for malarial experiments. PMID- 15830860 TI - [Preliminary study on the ecology of Trichobilharzia cercariae in the Huaihe river system]. AB - OBJECTIVE: To investigate the ecological habit and characteristics of Trichobilharzia cercariae in the Huaihe river system. METHODS: During June of 2002-May of 2003, Radix auricularia snails were collected monthly in Yaohe Fishery of the branch of the Huaihe River. The cercariae, released from the positive snails and collected by Shade Drop Bottle, were examined by direct press. Ducklings were infected by cercariae with different ages and examined for the infection by miracidia hatching method. RESULTS: The results showed a typical seasonal fluctuation in infection rate of the snails, i.e. the cercariae detection rate was 0.81% in July, 0.65% in June, 0.07%-0.26% in April, May, August, September and October respectively, with a significant difference (chi2 = 26.73, P < 0.01). The release of cercariae from the snails showed a diurnal pattern in the natural conditions, with a significant peak between 8:00-12:00. It was proven that the main factors affecting the emerging of cercariae were temperature and light. The cercariae showed positively phototaxis. It was also showed that 50% mortality of the newly released cercariae between 18-25 degrees C occurred at 41 h, and all cercariae died within 52 h. The infectivity of cercariae was age-dependent. Under 18-25 degrees C, the highest infectivity was found at 0.5-8 h after emergence, then declined steadily to negative by 40 h post emergence. CONCLUSION: The ecological characteristics of Trichobilharzia cercariae have been preliminarily verified in the Huaihe river system. PMID- 15830861 TI - [Discovering hookworms by gastroscopy from duodenal bulb in 4 cases]. PMID- 15830862 TI - [Study on surface adhesion protein 33 gene sequence of different Trichomonas vaginalis isolates]. AB - OBJECTIVE: To study genetic polymorphism of surface adhesion protein 33 (AP33) gene on the seven isolates of Trichomonas vaginalis. METHODS: PCR technique was performed to amplify AP33 gene from the seven isolates, DNA sequences were obtained from the AP33 gene of the isolates and phylogenetic tree was built. Minimal lethal concentrations (MLC) of metronidazole on the isolates were measured in vitro. RESULTS: Percentage of the similarity between 7 isolates and U87098 in GenBank was 98.2%-100%, which indicated a high homology and belonged to isotype isolates. There were four branches between Beijing 1 isolate and Tangshan isolate, Jiujiang 1 isolate and Jiujiang 2 isolate, Beijing 2 isolate and Jiujiang 3 isolate, Chengde isolate and U87098 isolate in phylogenetic tree, which showed a close genetic relationship respectively. No relativity was detected between geographical origin and genetic relationship. CONCLUSION: There is a close genetic relationship among the seven T. vaginalis isolates. MLC showed a difference between isolates which have close relationship. PMID- 15830863 TI - [The experimental study of garlicin in treating Pneumocystis carinii pneumonia in rats]. AB - OBJECTIVE: To assess the therapeutic effect of garlicin on Pneumocystis carinii pneumonia (PCP) of immunosuppressed rats. METHODS: The Wistar rats were injected intramuscularly continually with dexamethasone for eight weeks to establish the rat model of PCP. The rats were treated with garlicin, meanwhile control groups without treatment and with SMZ-TMP treatment were established respectively in PCP model. The efficacy was evaluated based on the lung weight, lung/body weight ratio and mean cyst number per 100 fields in lung print smear. RESULTS: The mean lung weight of the rats in garlicin is 1.73 +/- 0.17, lung/body weight ratio is 0.84 +/- 0.12, they are obvious lower than 2.31 +/- 0.35 and 1.25 +/- 0.35 of control (P <0.01). The numbers of cysts in lung print smear is reduced 62.9% compared with control. The results is similar to that with SMZ-TMP. CONCLUSION: Garlicin shows an obvious therapeutic effect on Pneumocystis carinii pneumonia in rats with an efficacy similar to that with SMZ-TMP. PMID- 15830864 TI - [Change of cytokines in mice with Echinococcus multilocularis infection]. AB - OBJECTIVE: To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection. METHODS: Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2, IFN gamma, TNF-alpha, IL-4, IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d. RESULTS: Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL 2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-alpha was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-gamma reached a peak after 80 d p.i., and decreased slowly after 140 d p.i. The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i. CONCLUSION: The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection. PMID- 15830865 TI - [Epitope vaccine: a new strategy for vaccine development]. PMID- 15830866 TI - [In vitro cultivation of Cryptosporidium and research progress of animal models]. PMID- 15830867 TI - [The evolution and development of "three criteria" for the diagnosis of neurocysticercosis]. PMID- 15830868 TI - [Clinical analysis on 50 cases of neurocysticercosis in children]. PMID- 15830869 TI - [Schistosome antibody detection by home microwave irradiation ELISA]. PMID- 15830870 TI - [Effects of the pre-microscopy processes on cellophane tape method for detecting pinworm eggs]. PMID- 15830871 TI - [Survey of Trichinella spiralis infection in pigs in Xiangfan city in 1993-2003]. PMID- 15830872 TI - [Fifteen years' surveillance of malaria after its transmission interrupted in Shandong Province]. PMID- 15830873 TI - [Prokaryotic expression of an antigen gene of Trichinella spiralis and identification of the recombinant protein]. AB - OBJECTIVE: To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein. METHODS: Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET-28c(+) expression vector and expressed in E. coli. Mice were immunized with the fusion protein incorporated into Freund' s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno-fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm. RESULTS: The fragment of Ts88 gene was expressed successfully in E. coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno-fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm. CONCLUSION: The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity. PMID- 15830874 TI - [A survey on the family Simuliidae (Diptera) at Zhenbao Island of Heilongjiang Province]. PMID- 15830875 TI - [Prediction of the impact of climate warming on transmission of schistosomiasis in China]. AB - OBJECTIVE: To predict the intensity and scale of impact on transmission of schistosomiasis japonica in China caused by the climate warming. METHODS: By using climate data from 193 weather stations in China from 1951 to 2000, the GIS database was created to analyze the tendency of average daily temperature. By using the results from the effective accumulated temperature models on Oncomelania snails and Schistosoma japonicum, the climate-transmission model for schistosomiasis was established at country level, by which the spatio-temporal analysis was performed to create the distribution maps of Oncomelania snails and Schistosoma japonicum, respectively, by means of GIS approaches based on the ratio of effective accumulated temperature to the snail or the parasite development temperature (ET/SDT) in all 193 stations. The potential distribution maps with the dispersal risk areas of schistosomiasis japonica in 2030 and 2050 were created based on forecast data that the average temperature of the country will increase by 1.7 degrees C in 2030 and by 2.2 degrees C in 2050. RESULTS: The GIS database of climate-schistosomiasis of the country was established. It was found that the average temperature in the last 5 decades inclined, especially after 1990 it increased significantly with its increasing regression formula T = 0.0198X - 28.476. The climate-transmission model for schistosomiasis was established, and it was found that the geographical distribution of Schistosoma japonicum was much larger than that of Oncomelania snails based on the ratio of ET/SDT. The prediction maps for distribution of schistosomiasis in 2030 and 2050 were created, respectively, which showed that the sensitive areas were extended with the time, the risk of expansion northward for schistosomiasis will be increasing due to directly the climate warming. CONCLUSION: It is predicted that a northward expansion of transmission area of schistosomiasis may occur due to the climate warming, the expanded potential area for schistosomiasis transmission will be important for future surveillance. PMID- 15830876 TI - Expression of DNA vaccine against Trichinella spiralis in Chinese hamster ovary cells. AB - OBJECTIVE: To observe the in vitro expression of DNA vaccine (recombinant eukaryotic expression plasmid pcDNA3-TspE1) encoding a Mr 31,000 antigen of Trichinella spiralis in Chinese hamster ovary (CHO) cells and analyze the antigenicity of the products expressed. METHODS: The recombinant plasmid pcDNA3 TspE1 was transfected into CHO cells by using cationic lipids (Lipofectamine 2000). The positive cell clones were screened by the selective antibiotic G418. The expressed products were identified by RT-PCR, IFAT, SDS-PAGE and Western blotting. RESULTS: The results of RT-PCR amplification showed that there was one band with 876 bp in CHO cells transfected with pcDNA3-TspE1 and no any bands in CHO cells transfected with the empty plasmid pcDNA3. The IFAT demonstrated that the pcDNA3-TspE1 transfected CHO cells reacted with sera from mice immunized with the recombinant fusion protein and from mice infected with T. spiralis, the bright yellow green fluorescence staining appeared in the transfected CHO cells. The pcDNA3 transfected and un-transfected CHO cells exhibited as orange color. The results of SDS-PAGE showed that there was one band with Mr 31,000 in culture supernatant of CHO cells transfected with pcDNA3-TspE1. Western blotting confirmed that the band with Mr 31,000 could be recognized by sera from mice immunized with the recombinant fusion protein, from rabbits immunized with T. spiralis muscle larval soluble antigens, from mice infected with T. spiralis and from patients with trichinellosis. Conclusion The mammalian CHO cells were transfected by the recombinant plasmid pcDNA3-TspE1, and the TspE1 gene of T. spiralis was expressed in the transfected CHO cells. The proteins expressed are secreted into cell culture supernatants and show the antigenicity of T. spiralis. PMID- 15830877 TI - [The clinical significance of interferon-gamma and its receptor in patients with schistosomiasis]. AB - OBJECTIVE: To study the expression of interferon-gamma (IFN-gamma) and type I interferon-y receptor (IFN-gammaRI) in the chronic and advanced patients of schistosomiasis japonica, and to discuss its clinical relationship with hepatic fibrosis. Methods IFN-yR1 in the peripheral blood lymphocytes was detected by ELISA, and the hyaluronic acid (HA) , collagen type IV (C-IV), procollagen type III (PC III), laminin (LN) were detected by radio-immunoassay (RIA) in schistosomiasis patients. The level of IFN-y, IFN-gammaR1 and serum markers of hepatic fibrosis were observed, and the relationship with each other was analyzed by statistical method. RESULTS: There was no difference in the expression of IFN gamma and IFN-gammaR1 between the patients with chronic schistosomiasis and the normal group (P > 0. 05 ), the IFN-gammaR1 in advanced cases without splenectomy was low (P <0. 05) , but IFN-y was high (P < 0. 01). The two indicators in the advanced schistosomiasis patients with splenectomy returned to normal. There was no corresponding relationship between the two indicators and HA, C-IV, PC III, LN with a r value of 0. 19, 0.20, 0. 14, and 0.21 respectively. CONCLUSION: There is a corresponding relationship between IFN-gamma and IFN-gammaR1; the expression of IFN-gammaR1 is related to the course of schistosomiasis, and the relationship with hepatic fibrosis needs further study. PMID- 15830878 TI - [Case report: a boy infected with Armillifer moniliformis]. PMID- 15830879 TI - [Analysis of cytochrome c oxidase I and cytochrome b genes of F1 in laboratory line Oncomelania hupensis hupensis]. AB - OBJECTIVE: To analyze the diversity of F1 in laboratory line Oncomelania hupensis hupensis. METHODS: Genomic DNA was isolated, and cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes were amplified by polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were analyzed by GENETYX-MAC software package (ver. 9). RESULTS: Pairwise divergences among six F1 individuals were found in 12.2% nucleotides in COI gene fragments. 13.5% of genetic divergences between O. h. hupensis and O. h. robertsoni were identified. 94 amino acids were observed in difference. In Cytb gene fragments, 6.4% of divergence was observed among four F1 samples. 25 amino acids were observed differently. The divergence of Cytb genes from O. h. robertsoni and O. h. hupensis were 13.6%, included 6 amino acids. CONCLUSION: Diversities were found in both COI and Cytb gene sequences of F1 in laboratory line O. h. hupensis. PMID- 15830880 TI - [Analysis of the sequences of the ribosomal DNA ITS2 region for differentiating anopheline mosquitoes within hyrcanus group]. AB - OBJECTIVE: To analyze the genetic characteristics of ribosomal DNA ITS2 region for differentiating anopheline mosquitoes within Hyrcanus group. METHODS: The ribosomal DNA ITS2 region of both laboratory line and filed collected An. anthropophagus and An. sinensis as well as the field collected An. yatsushiroensis and An. lesteri were amplified and sequenced. The sequencing data were then analyzed for the restriction mapping using Omega Sequencing analysis program. RESULTS AND CONCLUSION: The length of the sequences of An. sinensis, An. anthropophagus, An. yatsushiroensis and An. lesteri are 472, 452, 456 and 456 bp respectively. The restriction mapping showed that there were different restriction digesting sites among the ribosomal DNA ITS2 region sequences from An. sinensis, An. anthropophagus, An. yatsushiroensis and An. lesteri. On the basis of the sequence differences among the anopheline species within Hyrcanus group, it is possible to develop new technique for genetic identification of anopheline mosquitoes. PMID- 15830881 TI - [Change of lipid, esterase and lipase in mosquito larvae infected with Lagenidium giganteum]. AB - OBJECTIVE: To explore possible physiological and biochemical mechanisms of Lagenidium giganteum infection in killing mosquito larvae. METHODS: The content of lipid and the activities of esterase and lipase between the normal and infected mosquito larvae were observed with histochemical method. The results were photomicrograph and analysed by image analysis using computer. RESULTS: In 24 hrs after infection, the content of lipid in the infected Culex quinquefasciatus larvae was lower while the activities of esterase and lipase in the larvae were higher than the control. In 48 hrs and 78 hrs after infection, the content of lipid in the infected larvae of C. quinquefasciatus and Ae. albopictus were significantly lower while the activities of esterase and lipase were significantly higher than those of control. CONCLUSION: The content of lipid in the mosquito larvae decreased while the activities of esterase and lipase increased after the infection of Lagenidium giganteum. The disorder of lipid metabolism might be part of the killing mechanisms to C. quinquefasciatus and Ae. albopictus larvae. PMID- 15830882 TI - [Biological features of mouse macrophage transfected with Toxoplasma gondii GRA-1 gene]. AB - OBJECTIVE: To investigate the change of biological features of macrophages after transfected by Toxoplasma gondii GRA-1 genes (P24). METHODS: The transfected cells Cyto-P24-RAW264.7, Nuc-P24-RAW264.7, Mito-P24-RAW264.7 and ER-P24-RAW264.7 were studied by RT-PCR to determine the P24-mRNA expression. Growth features of the cells were examined with microscopy and the cell growth curve was developed. RESULTS: In four cell lines, expression of ER-P24-RAW 264.7 was found to be higher than the other three, and there was no P24-mRNA expression in either of the cells without P24 insert. The attachment and the proliferation of ER-P24 RAW264.7 were more rapid than normal RAW264.7. CONCLUSION: Transfection of mouse macrophages ER-RAW264. 7 strain with T. gondii P24 gene leads to a prominent change of biological features in the studied cell line. PMID- 15830883 TI - [Prevalence of anaphylaxis to dust mite in human population in Xuzhou]. AB - OBJECTIVE: To study the prevalence of anaphylaxis to dust mite in normal individuals and patients with different allergic disorders in Xuzhou. METHODS: By skin prick test (SPT) with Dermatophagoides farinae allergen extract, the prevalence of the normal individuals and OPD patients visiting hospital with allergic symptoms were examined for hypersensitivity to mite. Histamine equivalent criteria were used to evaluate the reactivity rate and the strength of skin response. Results 34 (15.3%) out of 222 pupils and students of different age groups from different levels of schools regarded as the normal population showed positive reaction to the mite allergen with usually only very weak response(+). There were no significant differences between the different age groups (P > 0.05), but it was different between 18.6% (22/118) in females and 11.5% (12/104) in males (P < 0.05). Also the reactivity rate of younger female pupils was higher than that of older ones. Among 515 cases from the OPD patients with allergic symptoms from pediatrics, ENT, respiratory and dermatology departments, 424 (82.3%) were positive with different grades of skin reaction. The rate of strong reactivity (> or = + + +) was 68.6% in allergic children, 63.5% in patients with allergic rhinitis, 41.9% in patients of respiratory department and 25.7% in patients with dermatitis. The allergic children of < or =10 years old showed highest rate of very strong reactivity (+ + + +), the rates declined along with the growing age. CONCLUSION: In Xuzhou area the SPT reactivity rate to dust mite for normal individuals was 15.3% with only weak response; while for allergic patients it was 82.3%, 47.9% of them showed strong and very strong responses. PMID- 15830884 TI - Molecular cloning and characterization of a Rac1 homologue cDNA from Trichomonas vaginalis. AB - OBJECTIVE: To clone and characterize a Rac1 homologue from Trichomonas vaginalis for studying cell cycle of the organism. METHODS: A cDNA library derived from T. vaginalis mRNA was constructed into lambda TriplEx2 phage vector. An expression sequence tag program was launched. Sequences of cDNA clones were analyzed using NCBI BLAST algorithms, and ClustalW and Treeview programs. RESULTS: A cDNA clone with a length of 714 base pairs was isolated. The sequence analysis showed that the cDNA clone has an open reading frame with 600 bp. The deduced amino acid sequence from the open reading frame contains 200 residuals and is most homologous to Rac1 subfamily of Rho GTPases with >60% identity. The conserved sequence elements of Rho GTPases, such as GTP-binding sites, GTPase-activating protein (GAP) interaction motifs, GTP-dissociation inhibitors (GDI) interaction motifs, guanine nucleotide exchange factor (GEF) interaction elements, etc, were detected in the amino acid sequence. The phylogenetic analysis showed that the cDNA clone is grouped in the Rac subfamily and is more closely related to Rac1 proteins of protozoa. CONCLUSION: The cDNA clone isolated belongs to Rac subfamily of Rho GTPases and is probably a Rac1 protein of T. vaginalis. PMID- 15830885 TI - [Cloning and sequence analysis of the ribosomal DNA ITS gene of Leishmania donovani isolates from hill foci of China]. AB - OBJECTIVE: To determine the nucleotide sequence of the ITS (internal transcribed spacer) gene of Leishmania donovani isolates from hill foci (L. d. SC10 and L. d. 6), and to find out the difference of the gene sequences between the two isolates. METHODS: Specific ITS fragments from nuclear DNA of two Leishmania isolates were amplified by PCR, cloned into pMD18-T vector, and finally sequenced by the dideoxy chain termination method. RESULTS: Sequence analysis showed that the amplified DNA fragments of the two isolates were 1027 bp ( L. d. SC10) and 1 028 bp (L. d. 6) respectively, showing a sequence difference. CONCLUSION: Sequence difference exists between the Leishmania isolates L. d. SC10 and L. d. 6 from hill foci in China. PMID- 15830886 TI - [Toxicity of alpha-terthienyl to the larvae of Aedes albopictus]. AB - OBJECTIVE: To investigate the toxicity of alpha-terthienyl to the larvae of Aedes albopictus, its influencing factors and effect on the larva development. METHODS: Under experimental ultraviolet-A (UVA), the number of dead,pupal or eclosive mosquito larvae was determined on the condition of different doses of alpha terthienyl and different disposal time in the dark; the number of dead larvae was also determined under sunlight on the condition of different doses of alpha terthienyl and different disposal time to water. RESULTS: The LC50 of alpha terthienyl to Aedes albopictus larvae was 2.37 microg/L under UVA. The best effect was shown when the larvae were incubated with alpha-terthienyl 3 h in dark. Alpha-terthienyl could significantly inhibit the larva development and the emergence of the pupae. Under strong sunlight, the larvae were quickly killed by high concentration alpha-terthienyl. The 24 hours effect of alpha-terthienyl was better when it was applied at 5 AM than that of at 10 AM and 1 PM. CONCLUSION: Alpha-terthienyl is an effective, practicable larvicide which prohibits the growth and development of the larvae of Aedes albopictus. PMID- 15830887 TI - [Studies on the genetic variation of two mitochondrial DNA molecules of Schistosoma japonicum]. AB - OBJECTIVE: To study the variation of Schistosoma japonicum through two mitochondrial DNA molecules. METHODS: Genomic DNA was isolated with kit, and the mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome c oxidase I (COI) gene fragments were amplified by polymerase chain reaction (PCR) and sequenced. The gene trees were constructed and the acquired data were analyzed with the help of bioinformatics. RESULTS: The gene trees showed that the Taiwan isolate and the mainland isolates can be divided in two groups: a group from the hilly region (Yunnan and Sichuan), another group from the lake region (Hunan, Jiangxi and Anhui); isolates from Hubei are at a different position on the gene trees. CONCLUSION: There are variations among the geographic isolates of Schistosoma japonicum in China, nevertheless, they have close kinship. PMID- 15830888 TI - [Molecular cloning of a protein antigen gene of Cysticercus cellulosae]. AB - OBJECTIVE: To immunoscreen one protein antigen gene from a cDNA library of Cysticercus cellulosae. METHODS: A eDNA library of C. cellulosae was constructed after cDNA was synthesized, and immunoscreened using rabbit anti-C. cellulosae polyclone antibody. The gene structure and its possible function were analyzed by comparing with the sequences available in the GenBank, after the insert of positive clone was subcloned to pBluescript SK plasmid and the nucleotide sequence of the cDNA was determined by dideoxynucleotide chain termination method using a Taq DyeDeoxy Terminator Cycle Sequencing Kit. The amino acid sequence was deduced from nucleotide sequence using GENETYX software. Homological search of the nucleotide sequences was done using BLAST in GenBank. RESULTS AND CONCLUSION: A cDNA clone of 1320 bp was isolated. The clone contained one open reading frame composed of 1260 bp encoding 420 amino acids, in which two potential glycosylation sites were found. The partial nucleotide sequence of the gene fragment showed high homology with the essential spectrin gene of Caenorhabditis elegans and the erythrocyte surface antigen gene of Plasmodium falciparum, when the gene fragment was homologically analyzed in GenBank. PMID- 15830889 TI - [Amplification, cloning and expression of 3-phosphoglycerate kinase gene from Clonorchis sinensis]. AB - OBJECTIVE: To construct a prokaryotic expression plasmid encoding 3 phosphoglycerate kinase (PGK) gene of Clonorchis sinensis and analyze its expression in E. coli JM109. METHODS: Total RNA was extracted with Trizol. The gene encoding PGK was amplified by RT-PCR from the total RNA, and was cloned into the pGEX-4T-1 vector. The recombinant plasmids pGEX-4T-1-PGK constructed were transferred into JM109, positive recombinants were screened and identified by tool enzyme digestion, PCR technique and sequencing. The recombinant was induced with IPTG to express the target protein. The expression product was characterized by SDS-PAGE. RESULTS: There were three clear bands from extracted total RNA, the PGK gene (1248 bp) was amplified by RT-PCR technique, the sequencing result was consistent with the known sequence. Ten clones were obtained by screening after transferring, six of which were positive recombinants. The positive recombinant was induced to express, and the SDS-PAGE showed the expression products was about Mr 70000. CONCLUSION: The recombination prokaryotic expression vector pGEX-4T-1 PGK has been reconstructed and a fused protein was expressed which contained Sj26 GST. PMID- 15830890 TI - [Phage display technique and its application in Toxoplasma gondii research]. PMID- 15830891 TI - [A case of facioliasis hepatica]. PMID- 15830892 TI - [Tribendimidine--a new broad-spectrum drug against intestinal helminths]. PMID- 15830893 TI - [The diagnosis of cerebral paragonimiasis in the active stage by CT and MRI]. PMID- 15830894 TI - [Re-staining of the slides of Entamoeba histolytica trophozoites]. PMID- 15830895 TI - [Technical research during the elimination of filariasis in Linyi City]. PMID- 15830896 TI - [Routine examination of leukorrhea in 3347 patients with trichomonal vaginitis]. PMID- 15830897 TI - [Report on surgical treatment of 3126 cases of liver cystic hydatidosis]. PMID- 15830898 TI - Latent growth curve analyses of parent influences on drinking progression among early adolescents. AB - OBJECTIVE: The nature of parent influences on early adolescent substance use was examined. METHOD: Latent growth curve analyses were used to examine data on a sample of 2,453 adolescents from seven middle schools who were randomized to a problem behavior prevention program or a control condition and were assessed a total of five times during sixth to ninth grade. RESULTS: Whereas the growth in the number of friends who drink was positively associated with adolescent drinking, parental involvement, monitoring and expectations over time provided direct protective effects against drinking progression and indirect effects by limiting increases in the number of friends who drink. CONCLUSIONS: The results provide evidence in a sample of early adolescents that parenting behavior- including involvement, monitoring and expectations--protected against progression in drinking directly as well as indirectly by limiting growth in the number of friends who drink. PMID- 15830899 TI - Parental alcohol use disorders and child delinquency: the mediating effects of executive functioning and chronic family stress. AB - OBJECTIVE: This study examines the relationship between parental alcohol use disorders (AUDs) and child violent and nonviolent delinquency. It also explores the mediating effects of executive functioning and chronic family stress on the parental AUD/child delinquency relationship. METHOD: Participants were 816 families with children (414 boys and 402 girls) born between 1981 and 1984 at Mater Misericordiae Mother's Hospital in Brisbane, Australia. Parents and children completed semistructured interviews, questionnaires and neuropsychological tests that assessed parental alcohol use, family psychiatric history, chronic family stress, child delinquency and child executive functioning. RESULTS: Paternal (but not maternal) AUDs predicted child violent and nonviolent delinquency. Executive functioning mediated the relationship between paternal AUDs and violent delinquency, whereas family stress mediated the relationship between paternal AUDs and both violent and nonviolent delinquency. CONCLUSIONS: Results support a biosocial conceptualization of the paternal AUD/delinquency relationship. They suggest that paternal AUDs may be associated with child executive functioning and family stress, which may in turn lead to child delinquency. PMID- 15830900 TI - A prospective investigation of relations between social influences and alcohol involvement during the transition into college. AB - OBJECTIVE: The present study used structural equation modeling to test whether prospective relations between prematriculation social influences and alcohol involvement in college were most consistent with peer selection, peer socialization or reciprocal determinism explanations and to determine if observed relations varied according to measurement interval. We tested the hypotheses that "active" (alcohol offers) and "passive" (social modeling, perceived norms) social influences would be uniquely and reciprocally associated with alcohol use and alcohol-related consequences across two and three waves of assessment. METHOD: Prospective undergraduates (N = 388) completed self-report assessments in the summer before matriculation (Wave 1), in the spring of their freshman year (Wave 2) and in the spring of their sophomore year (Wave 3). RESULTS: Reciprocal effects were observed between social influences and alcohol use in both two- and three-wave models. Some evidence was observed for reciprocal associations for social modeling with alcohol use and alcohol problems. Overall, however, only modest support was found for a reciprocal influence conceptualization of social influences in alcohol problems. For alcohol problems, the results were more consistent with selection effects. No significant reciprocal associations were observed for perceived norms. CONCLUSIONS: Findings generally support the Social Learning Theory concept of reciprocal determinism but suggest the relationship between individual drinking behaviors and the social environment varies when distinguishing between alcohol use and alcohol problems. These findings also point to the importance of distinguishing among different types of social influences when delineating processes that result from and lead to heavy drinking in college. PMID- 15830901 TI - Alcohol alters insulin-like growth factor-1 activated oct 2 POU domain gene expression in the immature female hypothalamus. AB - OBJECTIVE: The Oct 2 POU homeodomain gene has been shown to increase during late juvenile development; the upstream control of Oct 2 is not known, however. The insulin-like growth factor-1 (IGF-1) is known to act centrally to stimulate luteinizing hormone (LH)-releasing hormone (LHRH) release and advance female puberty. We therefore sought to determine if this peptide induces transcription of Oct 2 genes as an early pubertal event. Furthermore, as alcohol (ALC) blocks IGF-1-induced LHRH and LH release acutely, we aimed to determine if it could affect the ability of IGF-1 to stimulate Oct 2 gene expression. METHOD: Female rats, 25 days old, were administered saline or IGF-1 (rat IGF-1 20 ng/3 microI) in the third ventricle at 0900 hours and killed 2, 4 and 6 hours later for assessment of Oct 2 gene expression in the preoptic area (POA) and the medial basal hypothalamus (MBH). In another experiment, we determined whether ALC (3 g/kg) could block IGF-1-induced Oct 2 gene expression. RESULTS: In the POA, IGF-1 did not affect the expression of Oct 2a, but it increased the Oct 2c mRNA levels at 2 hours. In the MBH, both transcripts were elevated 4 hours after IGF-1 stimulation. ALC did not alter basal expression of either of the Oct 2 gene isoforms. In both regions, however, ALC blocked IGF-1-induced gene expression. CONCLUSIONS: IGF-1 induced Oct 2 genes prior to the normal increase during the late juvenile period, indicating this IGF- 1 induction may be an early event in the activation of the LHRH secretory pathway. ALC blocks this action, suggesting the Oct 2 POU gene is a likely target by which ALC can interfere with glial neuronal interactions and interrupt LHRH secretion during prepubertal development. PMID- 15830902 TI - Effect of ethanol on brain neuropeptides in adolescent and adult rats. AB - OBJECTIVE: Alcohol misuse early in life is associated with an increased risk of alcoholism. It is possible that this increased risk in adolescent drinkers is in part related to the susceptibility of the adolescent brain to ethanol. This study assessed the effects of ethanol exposure on several neuropeptides to begin to elucidate potential substrates that could mediate the differential effects of ethanol on adolescent and adult rats. METHOD: Male Sprague Dawley rats were exposed to ethanol vapor or air during adolescence (30 days old, n = 9, controls = 8) or adulthood (80-90 days old, n = 9, controls = 8) for 10 days. Blood alcohol concentrations averaging 250 mg/dl were maintained during this period. After 7 weeks of cessation from ethanol vapor, brain tissue was collected from the frontal cortex, caudate, hippocampus, amygdala and hypothalamus to assess the immunoreactivity levels of neuropeptide Y (NPY-LI), corticotropin-releasing hormone, substance P (SP-LI) and neurokinins (NK-LI). RESULTS: Ethanol exposure decreased overall hippocampal NPY-LI and increased SP-LI and NK-LI in the caudate, but these effects were more prominent in adult rats. Rats in the adult treatment groups (both ethanol exposed and controls) also had significantly lower levels of frontal cortical NK-LI, frontal cortical SP-LI and hypothalamic SP-LI relative to rats in the adolescent treatment groups. CONCLUSIONS: These data indicate that brief exposure to alcohol has long-term effects on levels of NPY LI, SP-LI and NK-LI. As these effects were primarily the result of changes in rats exposed to ethanol during adulthood, however, they are unlikely to contribute to the increased susceptibility of adolescents to the effects of chronic ethanol exposure. PMID- 15830903 TI - The role of Pavlovian cues in alcohol seeking in dependent and nondependent rats. AB - OBJECTIVE: Recent treatment approaches to substance use disorders have focused on reducing drug use by modifying drug-seeking behaviors in response to drug associated cues. Understanding the effect of alcohol-related stimuli on alcohol seeking responses is therefore of interest in the study of alcoholism. The present study examined the impact of ethanol- (ETOH) associated cues on selective ETOH-seeking behavior, using a Pavlovian-instrumental transfer design in groups of alcohol-dependent and nondependent rats. METHOD: Rats (N = 24) received Pavlovian conditioning in which each of two stimuli, a tone and white noise, was paired alternately with a 10% sweetened ETOH solution and a polycose-quinine solution. The rats were trained to perform two instrumental actions, with one action earning access to the sweetened ETOH and the other to the polycose quinine. After training, half of the animals were made ETOH-dependent by intragastric administration of 36 g/kg of ETOH over 4 days, whereas the remainder received intragastric administration of an isocaloric polycose solution. On the following day, subjects were given a choice extinction test in which they were free to choose between both actions with no outcomes being delivered. During this test, the ETOH- and polycose-associated Pavlovian cues were presented to assess performance of the two instrumental actions both in the presence and absence of these stimuli. RESULTS: Pavlovian cues associated with both the ETOH or the polycose exerted a nonspecific excitatory influence on reward-seeking behavior in both nondependent and alcohol-dependent rats. CONCLUSIONS: Responses through which rats gain access to ETOH appear to be subject to the general excitatory influence of the general motivational arousal induced by reward-related cues. It appears the rats' performance did not depend on encoding the specific consequences of their actions and thus was not affected by the selective retrieval or priming of those consequences in memory. PMID- 15830904 TI - Correlations among first-degree relatives for responses on the Self-Rating of the Effects of Alcohol Questionnaire in teenagers. AB - OBJECTIVE: The level of response (LR) to alcohol is an intermediate phenotype related to the alcoholism risk, with a heritability of at least 0.4 as estimated from alcohol challenge experiments. A measure of LR that can be used in adolescence at a time close to the first drinking experience, and that is less expensive than alcohol challenge experiments, is the Self-Rating of the Effects of Alcohol (SRE) Questionnaire. This questionnaire contains questions related to the number of drinks required for up to four different effects early in the drinking career ("first five" score). The familial characteristics of SRE responses have been estimated in adults; however, no study has evaluated familial and potential genetic components of the first five SRE score in adolescents. This article presents data regarding the familial nature of SRE-based scores among a sample of teenagers. METHOD: As part of the Collaborative Study on the Genetics of Alcoholism Phase II (follow-up) Protocol, SRE scores were available on 251 females and 236 males ages 13-19 years. These analyses compare the correlations among father-offspring, mother-offspring, sibling pairs and comparable unrelated individuals. RESULTS: For the 487 subjects, correlations among first-degree relatives ranged from 0.14 to 0.22 and were all significant. Correlations among comparable unrelated pairs ranged from 0.02 to 0.06 and were nonsignificant. When males and females were evaluated separately, the pattern of results, with higher correlations among first-degree relatives than among unrelated individuals, was similar, although, perhaps reflecting fewer subjects, correlations were more variable. CONCLUSIONS: Although not providing a definitive evaluation of heritability, the results are consistent with a potential proportion of the variance related to genes for first five SRE scores of between 0.3 and 0.4. These results parallel previously published data in adults and are similar to heritability estimates for LR on alcohol challenges. The data support the potential use of the first five SRE score in adolescents as a measure of LR in genetic and environmental model-based studies in young populations for whom the evaluation of LR is taking place at a time close to the onset of drinking. PMID- 15830905 TI - Depressive symptoms, DSM-IV alcohol abuse and their comorbidity among children of problem drinkers in a national survey: effects of parent and child gender and parent recovery status. AB - OBJECTIVE: The purpose of this study was to estimate the risk of depressive symptoms, DSM-IV alcohol abuse and their comorbidity among children of problem drinkers (COPDs) in college and the effects of parent and child gender and parent drinking status on outcomes and on COPDs' treatment utilization. METHOD: The study population was a nationally representative sample of 27,430 college students from 119 4-year colleges who completed survey questionnaires for the Harvard College Alcohol Study in 1997 and 1999. Logistic regression was used to estimate the psychiatric outcomes and patterns of treatment/counseling. RESULTS: COPDs who reported that their parents were active problem drinkers were at increased risk of depressive symptoms, DSM-IV alcohol abuse and their comorbidity. Female children of active problem drinkers (CAPDs) were at increased risk of depressive symptoms (odds ratio [OR] = 1.57,p < .01) and comorbidity of depressive symptoms and alcohol abuse (OR = 2.09, p < .01). Male CAPDs were at risk of depressive symptoms (OR = 1.69, p < .05) only. Stratified analysis by both parent and child gender revealed that depressive symptoms among female COPDs were affected by both paternal and maternal drinking, whereas among their male counterparts depressive symptoms were present only when the affected parent was the father. Male children of recovered problem drinkers were more likely to seek and receive psychiatric treatment/counseling than male CAPDs. CONCLUSIONS: The psychiatric risk of COPDs varied by respondent and parent gender, and by whether the affected parent was reported to be actively disordered or in recovery. The results highlight the importance of early and gender-specific interventions for COPDs. PMID- 15830906 TI - Predicting patterns of adolescent alcohol use: a longitudinal study. AB - OBJECTIVE: Because most studies of adolescent alcohol use have focused primarily on the frequency and quantity of consumption, we know little about how adolescent drinking patterns change during the high school years. The purpose of this article is to provide such data, as well as to identify some of the individual, family, social life and community predictors of changes in drinking patterns over time. METHOD: A sample of 1,253 students in grades 9 through 12 (57% female) in a large metropolitan school district participated. Three ethnicities were represented: African American, European American and Mexican American. Students completed questionnaires every 6 months for a 2-year period (n = 743 at Time 4). RESULTS: Cluster analyses of the drinking variables yielded one problem-drinking group (heavy, multiple-context drinking), two high-risk groups (i.e., date and outdoor drinking) and three normative groups (light, family/parent, moderate/ party drinking). The most predictable patterns of longitudinal changes in adolescent drinking were consistent with the following progression (or the reverse): abstainer --> normative drinker --> high-risk drinker --> problem drinker. Paternal attitudes toward adolescent drinking and peer involvement in antisocial behavior predicted movement into normative drinking; social activity with peers predicted movement into high-risk drinking; and emotional distress predicted the transition into problem drinking. CONCLUSIONS: These findings are consistent with the multistage social learning model, demonstrating that the predictors of adolescent alcohol use vary across different levels of adolescent alcohol involvement. PMID- 15830907 TI - The cumulative effects of acute alcohol consumption, individual differences and situational perceptions on sexual decision making. AB - OBJECTIVE: Past alcohol administration research has produced mixed findings regarding the role of acute alcohol consumption on sexual decision making. The purpose of this study was to evaluate a more complex theoretical model that places alcohol's acute effects in context, through the inclusion of background measures as well as affective and cognitive responses to the specific situation. METHOD: College students (90 men, 90 women) completed a survey that included measures of individual difference characteristics and past experiences; approximately 1 month later, they participated in an alcohol administration study. Participants were randomly assigned to one of three drink conditions (sober, placebo, alcohol), after which they read a story about a couple that wanted to have sex, but had no condoms available. RESULTS: In hierarchical multiple regression analyses, acute alcohol consumption significantly predicted participants' perceived likelihood that they would have sex without a condom in such a situation; an earlier step included gender, impulsivity, self-reported alcohol expectancies, frequency of heavy drinking, lifetime number of sexual partners and frequency of condom use. There was no significant effect associated with the expectancy that one had consumed alcohol. Neither was there a significant interaction between drink condition and self-reported alcohol expectancies. CONCLUSIONS: Through the inclusion of measures of individual differences and responses to the specific situation, this study provides a more nuanced understanding of the factors that affect college students' sexual decision making, compared with laboratory studies that examine the effects of acute alcohol consumption in isolation. Alcohol consumption explained a significant yet relatively small amount of variance. Researchers need to consider the broader context to understand how intoxication influences sexual decision making. PMID- 15830908 TI - Alcohol use in South Africa: findings from the first Demographic and Health Survey (1998). AB - OBJECTIVE: This study formed part of the 1998 South African Demographic and Health Survey, which included questions assessing the extent of alcohol use, risky drinking and alcohol problems among South Africans to obtain up-to-date baseline estimates of consumption and risky drinking and to inform intervention efforts. METHOD: A two-stage random sample of 13,826 persons ages 15 or older (59% women) was included in the survey. Alcohol use was assessed through eight questions, including the CAGE questionnaire. Frequency analyses for different age groups, geographic setting, education level, population group and gender were calculated, as were odds ratios for these variables in relation to symptoms of alcohol problems. RESULTS: Current alcohol consumption was reported by 45% of the men and 17% of the women. White men (71%) were most likely and Asian women (9%) least likely to be current drinkers. Urban residents were more likely than nonurban dwellers to report current drinking. One third of the current drinkers reported risky drinking over weekends, and 28% of the men and 10% of the women scored above the cutoff level on the CAGE questionnaire. Symptoms of alcohol problems were significantly associated with lower socioeconomic status, no school education in women and being older than 25 years of age. CONCLUSIONS: A comprehensive strategy is required to address the high levels of risky drinking and reported symptoms of alcohol problems. PMID- 15830909 TI - Immigration-associated variables and substance dependence. AB - OBJECTIVE: Drug-dependent patients show increased negative mood states such as anxiety and depression that interact with drug craving and quality of life. In this study, we compared immigrants with and without drug dependence and assessed whether drug dependence and social status after immigration contributed independently to negative mood states, stress-coping strategies and satisfaction with life in Germany. METHOD: Immigrants (N = 80) who had emigrated from the former Soviet Union to Germany were participants in the study. Drug-dependent immigrants (24 men, 16 women, mean [SD] age 33 [9] years) were compared with healthy immigrants (26 men, 14 women, mean age 35 [12] years) on current emotional status, trait-anxiety, depression, stress-coping strategies, drug craving and immigration-associated data. RESULTS: Compared with healthy immigrants, drug-dependent immigrants showed significantly higher levels of anxiety, depression and negative stress-coping strategies, independent of differences in education or employment status. Satisfaction with being in Germany was associated with both drug dependence and employment status. The severity of drug craving was significantly associated with negative mood states but not with any other measured sociodemographic variable. CONCLUSIONS: This study shows that drug dependence is associated with increased negative mood states and poor stress coping strategies, independent of the assessed sociodemographic variables. Drug dependence may thus severely interfere with acculturation in the host country. Because immigrants often face reduced treatment options as a result of cultural and language barriers, our findings underline the importance of adequate drug treatment facilities for dependent immigrants. PMID- 15830910 TI - Use of heavier drinking contexts among heterosexuals, homosexuals and bisexuals: results from a National Household Probability Survey. AB - OBJECTIVE: Extensive use of specific social contexts (bars and parties, for instance) by homosexuals and bisexuals is thought to be a factor in the higher rates of drinking among these groups. However, much of the empirical evidence behind these assumptions has been based on studies with methodological or sampling shortcomings. This article examines the epidemiological patterns of alcohol contexts in relation to sexual identity, using a large, national, probability population survey. METHOD: We used the 2000 National Alcohol Survey for these analyses. The prevalence of spending leisure time in each of two social contexts (bars and parties) that are associated with heavier drinking is examined by sexual orientation (heterosexual, homosexual, bisexual and self-identified heterosexuals with same sex partners). In addition, we compare levels of drinking within these contexts by sexual orientation within these groups. RESULTS: Exclusively heterosexual women spent less time in these two contexts relative to all other groups of women. Gay men spent considerably more time in bars compared with the other groups of men. Heterosexual women who reported same sex partners drink more at bars, and bisexual women drink more alcohol at both bars and parties than exclusively heterosexual women. For men, there were no significant differences for average consumption in any of these contexts. Entry of background and demographic variables into logistic regression analyses did little to modify these associations. CONCLUSIONS: There is empirical evidence that some groups of homosexual and bisexual women and men spend more time than heterosexual individuals in heavier drinking contexts. The frequency of being in these two social contexts does not appear to be associated with heavier drinking within these contexts for men, but it may be related to heavier drinking in those places among some groups of women. PMID- 15830911 TI - Reports of alcohol consumption and alcohol-related problems among homosexual, bisexual and heterosexual respondents: results from the 2000 National Alcohol Survey. AB - OBJECTIVE: Few population-based studies have explored differences in alcohol consumption by sexual orientation. This study examined the prevalence of abstinence, drinking, heavier drinking, alcohol-related problems, alcohol dependence and help-seeking among homosexual and bisexual women and men compared with heterosexuals. METHOD: Data are from the 2000 National Alcohol Survey, a national population-based survey of adults (N = 7,612), a Random Digit Dialing telephone survey of all 50 states of the United States and Washington, DC. Four categories of sexual orientation were created using questions on both sexual orientation self-identification and behavior: homosexual identified, bisexual identified, heterosexual identified with same sex partners and exclusively heterosexual. Five alcohol measures (past year) were used in the analyses: (1) mean number of drinks, (2) days consuming five or more drinks on a single occasion, (3) drunkenness, (4) negative social consequences (2 or more) and (5) Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, alcohol dependence. A lifetime measure of help-seeking for an alcohol problem was also analyzed. RESULTS: Few significant differences were found among men by sexual orientation. By contrast, both lesbians and bisexual women had lower abstention rates and significantly greater odds of reporting alcohol-related social consequences, alcohol dependence and past help-seeking for an alcohol problem. CONCLUSIONS: These findings suggest that alcohol dependence and alcohol-related consequences differ by sexual orientation, particularly among women. These findings also emphasize the need for the inclusion of sexual-orientation items in population-based surveys so that prevalence rates within these subgroups can be effectively monitored. PMID- 15830912 TI - Drinking to cope: a comparison of questionnaire and electronic diary reports. AB - OBJECTIVE: The purpose of this article is to assess the correspondence between questionnaire reports of dispositional drinking to cope (DTC) and different indices of daily DTC in a sample of nondependent heavy drinkers. METHOD: Data from electronic diary (ED) records of interpersonal problems, negative affect, coping and drinking were used to examine how questionnaire reports of dispositional DTC corresponded to ED reports of daily DTC and to within-person associations among ED reports of negative affect, interpersonal problems and drinking. In the current study, 98 community-residing heavy drinkers using EDs recorded moods, interpersonal problems, coping attempts, desire to drink and drinking for 21 consecutive days. RESULTS: The findings demonstrate correspondence between questionnaire reports of dispositional DTC and (1) ED reports of daily DTC, (2) intensity of negative mood associated with daily drinking initiation and (3) some between-person differences in within-day prediction of drinking and desire to drink from mood and interpersonal problem ratings. Those reporting higher dispositional DTC were somewhat more likely to drink in response to some negative moods and less likely to drink in response to some positive moods than those reporting relatively lower dispositional DTC, but these findings were mixed. CONCLUSIONS: Questionnaire reports of dispositional DTC are at best inconsistently linked to observed within-person associations among daily moods, interpersonal problems and drinking even among heavy drinkers. PMID- 15830913 TI - Using calculations to estimate blood alcohol concentrations for naturally occurring drinking episodes: a validity study. AB - OBJECTIVE: The purpose of this study is to determine the accuracy of retrospective estimated blood alcohol concentrations (eBACs) for re-creating intoxication resulting from a naturally occurring drinking event. This study identified five eBAC equations, applied them to self-report data and compared the results to actual blood alcohol concentration obtained by a breath test. METHOD: A convenience sample of 109 drinkers was recruited near drinking establishments and asked to provide breath alcohol concentration (BrAC) samples. Participants were contacted by telephone on the next waking day to gather data required for five algorithms that determine eBAC. BrAC and eBAC obtained from each equation were compared to determine the level of agreement between the two approaches. RESULTS: eBACs correlated highly with each other (r > or = 0.99); R2 for all algorithms ranged from 0.54 to 0.55 with BrAC as the criterion. On average, eBAC equations overestimated BrAC. Regression analysis identified the amount of time spent drinking, number of standard drinks, weight and year in school as factors related to discrepancy. CONCLUSIONS: These data indicate that, although all equations produce eBACs that are highly related, their relationship to BrAC does vary across equations. Using the best fitting equation, eBAC is more strongly correlated with BrAC when intoxication is less than 0.08 g/210 L of breath, and the magnitude of the relationship decreases as intoxication rises. PMID- 15830914 TI - Cross-lagged relationships between substance use and intimate partner violence among a sample of young adult women. AB - OBJECTIVE: The goal of this study was to examine the longitudinal relationship between substance use and intimate partner violence (IPV) victimization and perpetration among a sample of young adult women. METHOD: A sample of 509 women who participated in Waves 8 (age 23) and 9 (age 29) of a multiyear panel study and who indicated they were living with a partner or spouse at both time points provided the data for this investigation. Path analysis was used to examine the cross-lagged relationships between women's substance use and IPV victimization and perpetration over the two waves of data. RESULTS: Although strong within- and across-time associations between substance use and IPV victimization and perpetration were found at the bivariate level, substance use did not predict women's subsequent IPV victimization or perpetration in the cross-lagged model. Instead, victims of IPV at age 23 were found to be at an increased risk for later heavy drinking. Perpetrators of IPV at age 23 were less likely than nonperpetrators to report heavy drinking at age 29. CONCLUSIONS: The results suggest that substance use does not increase women's long-term risk of experiencing or perpetrating IPV but that victimization by IPV puts women at risk for subsequent heavy drinking. PMID- 15830915 TI - Free and total carnitine concentrations in pig plasma after oral ingestion of various L-carnitine compounds. AB - This study was undertaken to investigate the bioavailability of various L carnitine esters (acetyl-L-carnitine and lauroyl-L-carnitine) and salts (L carnitine L-tartrate, L-carnitine fumarate, L-carnitine magnesium citrate) relative to base of free L-carnitine. Six groups of five or six piglets each were administered orally a single dose of 40 mg L-carnitine equivalents/kg body weight of each of those L-carnitine compounds. A seventh group served as a control. Free and total plasma carnitine concentrations were determined 1, 2, 3.5, 7, 24, and 32 hours after administration of the single dose. Area-under-the-curve (AUC) values were calculated to assess the bioavailability of the L-carnitine compounds. AUC values, calculated for the time interval between 0 and 32 hours, for both free and total carnitine were similar for base of free L-carnitine and the three L-carnitine salts (L-carnitine L-tartrate, L-carnitine fumarate, L carnitine magnesium citrate) while those of the two esters (acetyl-L-carnitine, lauroyl-L-carnitine) were lower. Administration of L-carnitine L-tartrate yielded a higher plasma free carnitine AUC value for the time interval between 0 and 3.5 hours than administration of the other compounds. The data of this study suggest that L-carnitine salts have a similar bioavailability to that of free L-carnitine while L-carnitine esters have a lower one. The study also suggests that L carnitine L-tartrate is absorbed faster than the other L-carnitine compounds. PMID- 15830916 TI - Assessment of antioxidative activity of lipid- and water-soluble vitamins in human whole blood. Comparative analysis between a biological test and chemical methods. AB - Assessment of the antioxidant activity of vitamins and other compounds is of interest in the understanding of their in vivo effects. In this study, we have investigated the activity of several lipid and water-soluble vitamins in human whole blood. Measurements were carried out using a biological test that enables the evaluation of both red blood cells and plasma resistance against free radical activity induced by 2,2'-azobis (2-amidinopropane)hydrochloride (AAPH). Antioxidant activity of vitamins has been determined by using the biological test versus chemical methods (chemiluminescence, DMPD radical). We have observed strong anti-oxidant potentials for vitamins B6 and B9 with biological tests, but not with chemical methods. At 10 microM, the vitamin B9 efficiency in inhibiting radical-induced red blood cell hemolysis was almost three times higher than vitamin C efficiency and two times higher than alpha-tocopherol efficiency. Antioxidant activity was not observed for vitamins B1 or B2, nor for retinol. The weak activity of beta-carotene still remains to be investigated particularly in relation to oxygen pressure. Our study demonstrated that the biological test is more useful than the chemical methods employed in this instance, for the evaluation of antioxidant capacity of lipophilic and putatively biologically active compounds. PMID- 15830917 TI - Effect of increase of dietary micronutrient intake on oxidative stress indicators in HIV/AIDS patients. AB - Several recent studies in human immunodeficiency virus (HIV) patients have identified micronutrient deficiencies as affecting progression to acquired immunodeficiency syndrome (AIDS) and death. Although the mechanisms are not known, micronutrient deficiencies may exacerbate the oxidative stress induced by HIV. In addition, infection and its evolution likely lead to an increased requirement for nutritional micronutrients, especially antioxidants. To evaluate this, 40 relatively healthy, institutionalized HIV-infected individuals were recruited for assessment before or three months after fresh fruit and vegetable supply were increased due to seasonal supply. Seven-day dietary records were recorded at the beginning (December) and end of the three-month study period (March). Oxidative stress indices and CD4+, CD38+/CD8+, and CD95+ T-lymphocyte subsets were also measured at these times. No significant differences were found in calorie or protein intake across the study period, but vitamin A, C, and E intakes all increased. A number of redox indicators were modified (increase: total antioxidant status, glutathione peroxidase, and glutathione; and decrease: superoxide dismutase) during the study period. However, no change in malondialdehyde, hydroperoxides, or DNA damage was noted but a significant reduction in CD38+/CD8+ relative count was seen. Within the context and limitations of this study, the increase of dietary fruits and vegetables intake for three months had some beneficial effects on nutrition, systemic redox balance, and immune parameters in HIV-infected persons. PMID- 15830918 TI - Dietary intakes and blood concentrations of antioxidant vitamins in German vegans. AB - We report on an evaluation of intake and blood concentrations of antioxidant vitamins in a cross-sectional study of 104 German vegans, and examine the association between blood concentration of antioxidant vitamins and their intake as well as fruit and vegetable intake. Comparisons are made with dietary reference intakes and established threshold values (ETV) for blood concentrations. Mean intakes of vitamin C, E, and beta-carotene were higher than actual recommendations, but comparable to reported intakes in other vegan cohorts. Mean blood concentrations were 0.76 micromol/L for beta-carotene and 117 micromol/L for vitamin C, so that nearly all participants showed concentrations that were above the ETV for the prevention of chronic diseases. Although mean vitamin E intake was 24.8 mg/day, only 32% of female and 18% of male participants showed plasma concentrations above the ETV. However, the mean vitamin E/cholesterol ratio was high (5.97), indicating a good protection of low-density lipoprotein (LDL) against peroxidation. Of the reported antioxidant vitamins, only plasma vitamin C concentration was a good biomarker of fruit and vegetable intake. The high concentration of antioxidative compounds in plasma may be one of the reasons for the lower incidence of chronic diseases in people consuming a plant-only diet. PMID- 15830919 TI - Cardiovascular mortality and alcohol consumption in Russia. PMID- 15830920 TI - Jew's mellow leaves (Corchorus olitorius) suppress elevation of postprandial blood glucose levels in rats and humans. AB - The study was performed to explore the suppressive effect of Jew's mellow leaves (JML) on postprandial blood glucose levels in rats and humans. A soluble dietary fiber (SDF) was extracted from the freeze-dried JML powder. An elevation of the postprandial blood glucose level in rats given 1% or 2% JML-SDF solution orally together with 20% glucose solution was significantly suppressed as compared with that observed in the control rats given only glucose solution. When seven healthy young male adults ingested 225 mL of JML mixed juice containing 15 g of freeze dried powder with 75 g of glucose in the fasting state in the morning, the elevation of the postprandial blood glucose level was significantly suppressed as compared with the control subjects. The diffusion rate of glucose and the permeation rate of glucose in the cultured Caco-2 cells were both significantly reduced by the addition of appropriate amounts of JML-SDF when compared to the controls. These results indicate that the effective substance in JML for suppressing blood glucose elevation is a kind of mucilaginous SDF. The mechanism by which this suppression occurs may be largely attributable to the delayed absorption of glucose from the intestinal membrane in the upper digestive tract by viscous SDF. PMID- 15830921 TI - All-trans retinoic acid and glycated albumin reciprocally influence their effects in human mesangial cells. AB - All-trans-retinoic acid (tRA) modulates in human mesangial cells (MC) antioxidant defenses, the expression of interleukin-1beta-induced vascular cell-adhesion molecule-1 (VCAM-1), cyclooxygenase-2 (COX-2), and the retinoic acid-receptor beta (RAR-beta). The correlation of the serum levels of glycated hemoglobin A1c with tRA in type 2 diabetes mellitus patients led us to hypothesize that tRA and glycated albumin (GA), the main circulating glycated protein, might mutually interact in MC. We studied 1) the influence of tRA on GA effects in cultured MC: an assessment was made on how pre-incubation with tRA modified the effects of GA on intracellular oxidation and on the expression of mRNA and protein of COX-2 and VCAM-1; and 2) the influence of GA on tRA effects in MC: we studied how the induction of RARbeta expression by tRA was modified by GA. RESULTS: GA dose dependently increased intracellular oxidation and the expression of the molecules involved in leukocyte infiltration, namely COX-2 and VCAM-1 . Pre-incubation with tRA exacerbated GA effects by up to a three- to four-fold additional increase. In turn, induction by tRA of RAR-beta was fully inhibited by GA. Thus tRA and GA reciprocally influence their effects in MC. It is possible that this interaction may have a pathophysiological or pharmacological role in diabetic nephropathy. PMID- 15830922 TI - Supplementation with tomato-based products increases lycopene, phytofluene, and phytoene levels in human serum and protects against UV-light-induced erythema. AB - Carotenoids are suitable photoprotectants, and beta-carotene supplements are used for protection against ultraviolet (UV) light-induced erythema. Protective effects are also observed when carotenoids are provided with the diet. Here, we investigated the photoprotective effects of synthetic lycopene in comparison with a tomato extract (Lyc-o-Mato) and a drink containing solubilized Lyc-o-Mato (Lyc o-Guard-Drink). With these different sources, the volunteers ingested similar amounts of lycopene (about 10 mg/day). After 12 weeks of supplementation, significant increases in lycopene serum levels and total skin carotenoids were observed in all groups. Significant increases in the serum levels of phytofluene and phytoene occurred in the Lyc-o-Mato and the Lyc-o-Guard-Drink group. At weeks 0, 4, and 12 an erythema was induced with a solar light simulator. Dorsal skin of each subject was irradiated with 1.25 minimal erythemal dose (MED). Reddening of the skin was evaluated before and 24 hours after irradiation by chromametry and expressed as positive a-values (red/green-axis). delta a-values (difference of a value before irradiation and after 24 hours) were used as an index of erythema intensity. A decrease in the delta a-value from week 0 to week 12, indicating prevention of erythema formation, was observed in all groups. Compared to week 0, the delta a-value at week 12 was 25% lower in the synthetic lycopene group. The protective effect was more pronounced in the Lyc-o-Mato (38%) and Lyc-o-Guard Drink (48%) groups. In the two latter groups, phytofluene and phytoene may have contributed to protection. Both of these carotenoids exhibit absorption maxima at wavelengths of UV light. Absorption of UV light protects skin from photodamage and might explain the differences observed between groups. PMID- 15830923 TI - Antioxidant and anti-atherogenic activities of olive oil phenolics. AB - The aim of the current study was to investigate the antioxidant and cellular activity of the olive oil phenolics oleuropein, tyrosol, hydroxytyrosol, and homovanillic alcohol (which is also a major metabolite of hydroxytyrosol). Well characterized chemical and biochemical assays were used to assess the antioxidant potential of the compounds. Further experiments investigated their influence in cell culture on cytotoxic effects of hydrogen peroxide and oxidized low-density lipoprotein (LDL), nitric oxide production by activated macrophages, and secretion of chemoattractant and cell adhesion molecules by the endothelium. Inhibitory influences on in vitro platelet aggregation were also measured. The antioxidant assays indicated that homovanillic alcohol was a significantly more potent antioxidant than the other phenolics, both in chemical assays and in prolonging the lag phase of LDL oxidation. Cell culture experiments suggested that the olive oil phenolics induce a significant reduction in the secretion of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 (and a trend towards a reduced secretion of monocyte chemoattractant protein-1), and protect against cytotoxic effects of hydrogen peroxide and oxidized LDL. However, no influence on nitric oxide production or platelet aggregation was evident. The data show that olive oil phenolics have biochemical and cellular actions, which, if also apparent in vivo, could exert cardioprotective effects. PMID- 15830924 TI - Moderate exercise combined with dietary vitamins C and E counteracts oxidative stress in the kidney and lens of streptozotocin-induced diabetic-rat. AB - Oxidative stress has a key role in the pathogenesis of diabetes-induced cataract formation and nephropathy. Daily moderate exercise and vitamins C and E (VCE) supplementation can be beneficial to diabetes due to reducing blood glucose and free radical production. The aim of this study was to analyze the effect of moderate exercise with vitamin VCE on lipid peroxidation (LP) and antioxidative systems in the kidneys and lens of streptozotocin-induced diabetic rats. Forty female Wistar rats were used. They were randomly divided into four groups. The first and second groups were used as control and diabetic groups. The third group was the diabetic-exercise group. VCE-supplemented feed was given to diabetic exercise rats constituting the fourth group. Animals in the exercised groups were moderately exercised daily on a treadmill for three weeks (five days a week). Diabetes was induced on day zero of exercise. Body weights in the four groups were recorded weekly. Lens and kidney samples were taken from all animals on day 20. Glutathione peroxidase (GSH-Px), reduced glutathione (GSH), vitamin E, and beta-carotene levels in kidney and lens, albumin in plasma, and body weight were significantly lower in the diabetic group than in the control group, whereas there was a significant increase in LP of kidney and lens as well as plasma glucose, urea, and creatinine levels in the diabetic group. The decrease in antioxidant enzymes, vitamins, and albumin and the increase in LP and glucose levels in diabetic rats were significantly improved with exercise and VCE supplementation. In the diabetic animals, the decreased beta-carotene and vitamins A levels in kidney did not improve through exercise only, although their levels were increased by exercise plus VCE supplementation. In conclusion, these data demonstrate that lipid peroxidation increases in the lens and kidney of diabetic animals and this could be due to decreases in antioxidant vitamins and enzymes. However, dietary VCE with moderate exercise may strengthen the antioxidant defense system through the reduction of ROS and blood glucose levels. The VCE supplementations with exercise may play a role in preventing the development of diabetic nephropathy and cataract formation in diabetic animals. PMID- 15830925 TI - Determination of odour-causing volatile organic compounds in cork stoppers by multiple headspace solid-phase microextraction. AB - Multiple headspace solid-phase microextraction (MHS-SPME) coupled with gas chromatography-mass spectrometry has been applied in order to determine 2,4,6 trichloroanisole (2,4,6-TCA), guaiacol, 1-octen-3-ol and 1-octen-3-one in three samples of cork stoppers. These compounds are responsible for cork taint in wine and can modify the organoleptic properties of bottled wine. Variables such as temperature, addition of water, extraction time, and amount of cork were studied. The extractions were performed with a 50/30 microm divinylbenzene-carboxen polydimethylsiloxane (DVB-CAR-PDMS) fibre for 45 min at 100 degrees C using 20 mg of cork. For calibration, 50 microL of VOC aqueous solutions were used and the extraction were carried out for 45 min at 75 degrees C. The limits of detection of the method expressed as ng of VOC per g of cork were 0.3 for 2,4,6-TCA, 7.5 for guaiacol, 1.7 for 1-octen-3-one and 1.9 for 1-octen-3-ol. Relative standard deviation of replicate samples was less than 10%. Significant losses of analytes were observed when the samples were ground at room temperature. Finally, a recovery study was performed and the MHS-SPME results were validated using Soxhlet extraction results. PMID- 15830926 TI - Assay of free ferulic acid and total ferulic acid for quality assessment of Angelica sinensis. AB - Activity of Chinese Danggui (DG), the processed root of Angelica sinensis (Oliv.) Diels, is linked to the ferulic acid content but the stability of ferulic acid during extraction for medicinal use is not known. The stabilities of ferulic acid and coniferyl ferulate were evaluated in the extracts of DG using a variety of extraction solvents. These included various combinations and proportions of methanol, water, formic acid, 1 M aqueous hydrochloric acid and 2% sodium hydrogen carbonate (NaHCO3) in water. Coniferyl ferulate was found liable to hydrolyze into ferulic acid in neutral, strongly acidic and basic solvents, where heat and water could facilitate this hydrolysis. However, the hydrolysis was relatively resisted in weakly organic acid. Based on the stability evaluation, two new terms, namely: free ferulic acid and total ferulic acid, were suggested and defined. Free ferulic acid refers to the natural content of ferulic acid in herbs. Total ferulic acid means the sum of free ferulic acid plus the amount of related hydrolyzed components. Meanwhile, the high-performance liquid chromatographic (HPLC) method was developed to assay free ferulic acid and total ferulic acid in DG using methanol-formic acid (95:5) and methanol-2% NaHCO3 in water (95:5) as extraction solvents, respectively. Ten DG samples were investigated on their contents of free and total ferulic acid. The results indicated that the amount variety of free ferulic acid was larger than that of their counterparts, and the ratio of total ferulic acid to free ferulic acid was 4.07 +/- 2.73 (mean +/- SD, n = 10). The chemical assay of DG using total ferulic acid content would be a better choice to assess the herbal quality and was recommended. PMID- 15830927 TI - Analysis of persistent organic pollutants in marine sediments using a novel microwave assisted solvent extraction and liquid-phase microextraction technique. AB - A simple and novel analytical method for quantifying persistent organic pollutants (POPs) in marine sediments has been developed using microwave assisted solvent extraction (MASE) and liquid-phase microextraction (LPME) using hollow fibre membrane (HFM). POPs studied included twelve organochlorine pesticides (OCP) and eight polychlorinated biphenyl (PCB) congeners. MASE was used for the extraction of POPs from 1 g of sediment using 10 ml of ultrapure water at 600 W for 20 min at 80 degrees C. The extract was subsequently subjected to a single step LPME-HFM cleanup and enrichment procedure. Recovery varied between 73 and 111% for OCPs; and 86-110% for PCBs, and exceeded levels achieved for conventional multi-step Soxhlet extraction coupled with solid-phase extraction. The method detection limit for each POP analyte ranged from 0.07 to 0.70 ng g( 1), and peak areas were proportional to analyte concentrations in the range of 5 500 ng g(-1). Relative standard deviations of less than 20% was obtained, based on triplicate sample analysis. The optimized technique was successfully applied to POP analysis of marine sediments collected from the northeastern and southwestern areas of Singapore's coastal environment. PMID- 15830928 TI - Simultaneous microwave-assisted extraction of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, phthalate esters and nonylphenols in sediments. AB - A new method was developed for the simultaneous extraction of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), phthalate esters (PEs), nonylphenols (NPs) and nonylphenol mono- and diethoxylates (NP1EOs and NP2EOs, respectively) in sediment samples by means of a closed microwave system. The extractions were carried out at 21 psi and 80% of microwave power and 15 ml of acetone were used as the common extraction solvent. The filtered extract was further fractionated in two groups using Florisil cartridges: PAHs and PCBs were eluted with n-hexane:toluene (4:1) and the PEs, NPs and ethoxylates were eluted with ethyl acetate. All the compounds were analysed by gas chromatography-mass spectrometry (GC-MS). In case of PAHs and PCBs, the developed method was validated by comparison of the results obtained for the certified reference material NIST 1944 with the certified values. In the absence of a reference material for phthalate esters and nonylphenols, one sediment sample was extracted twice under the optimal conditions in order to check than an exhaustive extraction of the analytes occurred. This method is currently used in the study of the distribution of those organic contaminants in the estuaries of the Bay of Biscay (Spain). PMID- 15830929 TI - Development of a total analytical system by interfacing membrane extraction, pervaporation and high-performance liquid chromatography. AB - This paper discusses the interfacing of continuous membrane extraction, pervaporation and on-line HPLC-UV detection into a total analytical system (TAS). Organics from a water sample were extracted into an organic solvent, and then concentrated via pervaporation prior to HPLC-UV detection. Factors affecting the system performance were studied. With optimized experimental parameters enrichment factors as high as 192 were obtained, the method detection limits were at low ng/mL levels, and the precisions were better than 5%. PMID- 15830930 TI - Direct separation of regio- and enantiomeric isomers of diacylglycerols by a tandem column high-performance liquid chromatography. AB - A novel HPLC-based method for direct separation of the three isomers of mono-acid diacylglycerols (DAGs), i.e., 1,2-DAG, 2,3-DAG and 1,3-DAG, has been established. The method employs a tandem column system, in which two different columns (a conventional silica gel column and a chiral stationary phase column) are connected in series. Two isomeric mixtures of DAGs (i.e., dicapryloylglycerol and dioleoylglycerol) and lipase-catalyzed reaction mixtures were successfully resolved on the tandem column HPLC system without any derivatization prior to the analysis. According to the established analytical method, stereoselectivity of two lipases toward mono-acid triacylglycerols in ethanolysis reaction was investigated. The tested enzymes were immobilized Candida antarctica lipase B (CALB) and Rhizomucor miehei lipase (RML). Analyses of the enantiomeric purity of 1,2-DAG and 2,3-DAG, generated as intermediates during the reaction, revealed that CALB and RML have sn-3 and sn-1 stereopreference, respectively. PMID- 15830931 TI - Distribution analysis of ultra-high molecular mass poly(ethylene oxide) containing silica particles by size-exclusion chromatography with dual light scattering and refractometric detection. AB - Two different size-exclusion chromatography (SEC) systems, connected in-line either to a low-angle light scattering (LALS) or to a multiangle light scattering (MALS) detector, are employed for determination of molecular mass distributions (MMD) of poly(ethylene oxide) (PEO) samples having a weight average molecular mass up to eight millions. The detrimental effect of the presence of strongly scattering silica particles in the samples on the light scattering signal can be eliminated using a suitable sample dissolution procedure utilizing silica solubility in aqueous mobile phase. The selection of flow-rate and sample concentration have a large impact on the obtained results. Hydrodynamic retardation phenomena and nonlinearity effects are shown to introduce severe errors in the molecular mass distributions unless flow-rate and sample concentration are kept at sufficiently low levels. Self-compensating ability of the dual detection in flow-rate effects is shown to be the main advantage here. A good agreement between the results obtained using LALS and MALS detection is found provided that a carefully selected angular extrapolation procedure is used in the case of MALS data. Thus, using carefully selected experimental conditions, SEC with light-scattering (LS) and refractometric detection proved to be an efficient technique for MMD characterisation also of ultra-high molecular mass (UHM) PEO polymers. PMID- 15830932 TI - Elimination of edge effects in micro-thermal field-flow fractionation channel of low aspect ratio by splitting the carrier liquid flow into the main central stream and the thin stream layers at the side channel walls. AB - An optimized construction of the separation channel for micro-thermal field-flow fractionation (FFF) was proposed and studied experimentally. The sample is injected in such a manner that its zone moves along the channel only in the main central stream where the flow velocity profile in the plane parallel to the main accumulation wall is practically flat. This central stream is separated from the contact with the side walls of the channel by thin flowing layers of the free carrier liquid. The retained species do not reach the thin liquid streams at the side walls where the flow rate decreases rapidly to achieve zero at the side wall according to the established 3D flow velocity profile. Such a construction of the channel allows one to reduce the aspect ratio (the ratio of the channel breadth b to its thickness w) without increasing the zone broadening. The hydrodynamic splitting of the outlet streams allows one not only to increase the concentration of the detected species but also the determination of the sign of Soret coefficient. PMID- 15830933 TI - Optimization of parameters of high-performance displacement chromatography for separation of soybean phosphatidylcholine and phosphatidylethanolamine. AB - Hundred milligrams of soybean phospholipids were successfully separated by using high-performance displacement chromatography (HPDC) on a 150mm x 4.6mm analytical silica column (3-5 microm packings) with dichloromethane-methanol (9:1, v/v) as carrier and ethanolamine as displacer. From the viewpoint of preparative separation, the effects of loading amount, concentration and flow-rate of displacer on separation efficiency were investigated using throughput and recovery as indices. The parameters were optimized by orthogonal test design and statistical analysis method. Under the optimum conditions, namely displacer concentration being 167 mM, the flow-rate of displacer at 0.2 ml/min and concentration of sample being 211 mg/ml (factual loading amount 211 mg/ml x 0.7 ml = 148 mg), the purity, throughput and recovery of obtained soybean phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were 80.2%, 65.7 mg/h, 70.9% and 90.5%, 272.6 mg/h, 88.3%, respectively. In addition, selections of regenerant and appropriate regeneration condition were also studied. PMID- 15830934 TI - New approach to linear gradient elution used for optimisation in reversed-phase liquid chromatography. AB - A new mathematical treatment concerning the gradient elution in reversed-phase liquid chromatography when the volume fraction psi of an organic modifier in the water-organic mobile phase varies linearly with time is presented. The experimental ln k versus psi curve, where k is the retention factor under isocratic conditions in a binary mobile phase, is subdivided into a finite number of linear portions and the solute gradient retention time tR is calculated by means of an analytical expression arising from the fundamental equation of gradient elution. The validity of the proposed analytical expression and the methodology followed for the calculation of tR was tested using eight catechol related solutes with mobile phases modified by methanol or acetonitrile. It was found that in all cases the accuracy of the predicted gradient retention times is very satisfactory because it is the same with the accuracy of the retention times predicted under isocratic conditions. Finally, the above method for estimating gradient retention times was used in an optimisation algorithm, which determines the best variation pattern of psi that leads to the optimum separation of a mixture of solutes at different values of the total elution time. PMID- 15830935 TI - Evaluation of gas chromatography-tandem quadrupole mass spectrometry for the determination of organochlorine pesticides in fats and oils. AB - A gas chromatography-tandem quadrupole mass spectrometry multi-residue method for the analysis of 19 organochlorine pesticides in fats and oils has been developed. Gel permeation chromatography was employed to remove lipid material prior to GC MS/MS analysis. Average recoveries of the pesticides spiked at 10 and 50 microg kg(-1) into fish oil, pork fat, olive oil and hydrogenated vegetable oil were typically in the range 70-110% with relative standard deviations generally less than 10%. Calculated limits of detection are between 0.1 and 2.0 microg kg(-1) and results obtained for the analysis of proficiency test materials are in good agreement with assigned values. The higher selectivity of the GC-MS/MS compared to electron capture detection and GC-MS in selective ion monitoring mode allowed unambiguous identification and confirmation of all the target pesticides at low microg kg(-1) levels in fats and oils in a single analysis. PMID- 15830936 TI - Determination of thermodynamic properties of isotactic poly(1-butene) at infinite dilution using density and inverse gas chromatography. AB - The partial molar volumes, V1(M), and the molar volume of isotactic crystalline low-molecular-weight poly(1-butene), iPBu-1, V1, have been calculated from the measured density of {iPBu-1 + solvent (n-hexane, n-heptane, n-nonane, n-decane, p xylene, cyclohexane and chloroform)} systems. Some of the thermodynamic quantities were also obtained for the iPBu-1 with eight hydrocarbons (n-octane, n decane, n-undecane, n-dodecane, n-tridecane, o-xylene, m-xylene, p-xylene) by the method of inverse gas chromatography at various temperatures. The weight fraction activity coefficients of the solvent at infinite dilution, omega2(infinity) and the Flory-Huggins thermodynamic interaction parameters, chi21(infinity), between polymer and solvents were determined. The partial molar free energy, deltaG2(infinity), the partial molar heat of mixing, deltaH2(infinity), at infinite dilution and the polymer solubility parameter, delta1, were calculated. Additionally, the (solid + liquid) binary mixtures equilibria, SLE, of iPBu-1 with three hydrocarbons (n-octane, n-decane and m-xylene) were studied by a dynamic method. By performing these experiments over a large concentration range, the T-x phase diagrams of the polymer-solvent systems were constructed. The excess Gibbs energy models were used to describe the nonideal behaviour of the liquid phase. The omega2(infinity) were determined from the solubility measurements and were predicted by using the UNIFAC FV model. PMID- 15830937 TI - Chemometric studies of retention in capillary gas chromatographic separation of hydrocarbons in coupled columns. AB - This paper describes how different multivariate analysis and classification methods can be used, to characterize the gas chromatographic separation of complex hydrocarbon mixtures in three columns coupled in series. Principal component analysis (PCA), correspondence factor analysis (CFA), and hierarchical ascending classification (HAC) were used as potential tools for evaluating the experiments on single columns and on column series. It has been demonstrated that: (1) multivariate analysis with PCA and CFA offers a powerful strategy to search for the main factors influencing the separation of hydrocarbons without a priori knowledge of the key factors of the separation. (2) With CFA the contribution of retention due to vapour pressure can be minimized. The use of retention indices, which use the n-alkanes as reference compounds, also helps to decrease the dominant focus on vapour pressure in favor of the more selectivity based interaction forces. (3) CFA helps to analyze the degree of relevance of the chosen experimental design to the most important factors, controlling chromatographic selectivity. PMID- 15830939 TI - Isotachophoretic determination of carboxylic acids in biodegradation samples. AB - In the current study a method of isotachophoretic separation of selected carboxylic acids was developed. The method was used for the determination of carboxylated oligo(ethylene glycol)s and their degradation products in biodegradation tests of PEG 250 DA [a mixture of dicarboxylated oligo(ethylene glycol)s]. Two tests were performed in the studies: the Organization for Economic Cooperation and Development (OECD) screening test and the river water die-away test. Both the biodegradation tests proved relatively fast biodegradation of the studied compounds. In the OECD screening test the biodegradation was faster than in the river water die-away test which can be ascribed to a higher concentration of bacteria in the biodegradation liquor. The minimal sample pretreatment and relatively low cost of analysis by the isotachophoretic method used here make it a good alternative to existing methods of carboxylic acids analysis. PMID- 15830938 TI - Analysis of chemical warfare agents. II. Use of thiols and statistical experimental design for the trace level determination of vesicant compounds in air samples. AB - Thermal desorption with gas chromatography-mass spectrometry (TD-GC-MS) remains the technique of choice for analysis of trace concentrations of analytes in air samples. This paper describes the development and application of a method for analysing the vesicant compounds sulfur mustard and Lewisites I-III. 3,4 Dimercaptotoluene and butanethiol were used to spike sorbent tubes and vesicant vapours sampled; Lewisite I and II reacted with the thiols while sulfur mustard and Lewisite III did not. Statistical experimental design was used to optimise thermal desorption parameters and the optimum method used to determine vesicant compounds in headspace samples taken from a decontamination trial. 3,4 Dimercaptotoluene reacted with Lewisites I and II to give a common derivative with a limit of detection (LOD) of 260 microg m(-3), while the butanethiol gave distinct derivatives with limits of detection around 30 microg m(-3). PMID- 15830940 TI - Chiral separation with novel (S)-biotin-bonded silica gel for liquid chromatography. AB - Liquid chromatographic separation of enantiomers was accomplished using a chiral stationary phase (CSP) derived from (S)-biotin on silica gel. In both nonaqueous and aqueous media, this CSP (1) permitted separation of racemic amino acid derivatives based on hydrogen bonding with a urea moiety of the biotin moiety. PMID- 15830941 TI - Purification and characterization of a solvent stable protease from Pseudomonas aeruginosa PseA. AB - A solvent tolerant Pseudomonas aeruginosa PseA strain was isolated from soil. It secreted a novel alkaline protease, which was stable and active in the presence of range of organic solvents, thus potentially useful for catalysis in non aqueous media. The protease was purified 11.6-fold with 60% recovery by combination of ion exchange and hydrophobic interaction chromatography using Q Sepharose and Phenyl Sepharose 6 Fast Flow matrix, respectively. The apparent molecular mass based on the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was estimated to be 35,000 Da. The enzyme was stable in the pH range of 6.0-9.0, the optimum being 8.0. The Km and Vmax towards caseinolytic activity were found to be 2.7 mg/ml and 3 micromol/min, respectively. The protease was most active at 60 degrees C and characterized as a metalloprotease because of its sensitivity to EDTA and 1,10-phenanthroline. It was tested positive for elastase activity towards elastin-orcein, thus appears to be an elastase, which is known as pseudolysin in other strains of P. aeruginosa. The protease withstands range of detergents, surfactants and solvents. It is stable and active in all the solvents having log P above 3.2, at least up to 72 h. These two properties make it an ideal choice for applications in detergent formulations and enzymatic peptide synthesis. PMID- 15830942 TI - Ultrathin phenyl-functionalized solid phase microextraction fiber coating developed by sol-gel deposition. AB - A new sol-gel application for the development of SPME fibers is described. Phenyltrimethoxysilane (PTMOS) and methyltrimethoxysilane (MTMOS) were the sol gel precursors used at different proportions, together with different water contents, catalyst and reaction time. It was observed that obtaining a good film quality was determinant for a good extracting fiber performance. The film thickness ranged 0.2-1 microm and could not be increased by multi-coating processes. Apparently, a dense, non-porous microstructure was obtained. These coatings exhibited a strong hydrophobic character, as shown by the capability of extraction of long chain and apolar aromatic compounds, which, was comparable to that of the 100 microm polydimethylsiloxane (PDMS) and 65 microm carbowax divinylbenzene (CW-DVB). The developed fiber has shown high thermal (350 degrees C) and organic solvent stability (ethanol, toluene and dichloromethane), thus bearing adequate characteristics to be associated to GC and potentialities that may also envisage suitability for HPLC. The new fibers may be useful for the microextraction of non-polar compounds, although at trace levels and in simple matrixes only, due to the susceptibility to competition. PMID- 15830943 TI - Evaluation of new selective molecularly imprinted polymers prepared by precipitation polymerisation for the extraction of phenylurea herbicides. AB - Three different molecularly imprinted polymers (MIPs) have been prepared by precipitation polymerisation using linuron (LIN) or isoproturon (IPN) (phenylurea herbicides) as templates and methacrylic acid (MAA) or trifluormethacrylic acid (TFMAA) as functional monomers. The ability of the different polymers to selectively rebind not only the template but also other phenylurea herbicides has been evaluated. In parallel, the influence of the different templates and functional monomers used during polymers synthesis on the performance of the obtained MIPs was also studied through different rebinding experiments. The experimental binding isotherms were fitted to the Langmuir-Freundlich isotherm allowing to describe the kind of binding sites present in the imprinted polymers under study. It was concluded that TFMAA-based polymer using IPN as template presents the best properties to be used as a selective sorbent for the extraction of phenylurea herbicides. PMID- 15830944 TI - Determination of pesticide residues in olives and olive oil by matrix solid-phase dispersion followed by gas chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. AB - A novel analytical approach has been developed and evaluated for the quantitative analysis of a selected group of widely used pesticides (dimethoate, simazine, atrazine, diuron, terbuthylazine, methyl-parathion, methyl-pirimiphos, endosulfan I, endosulfan II, endosulfan sulphate, cypermethrin and deltamethrin), which can be found at trace levels in olive oil and olives. The proposed methodology is based on matrix solid-phase dispersion (MSPD), (with a preliminary liquid-liquid extraction in olive oil samples) using aminopropyl as sorbent material with a clean-up performed in the elution step with Florisil, followed by mass spectrometric identification and quantitation of the selected pesticides using both gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode and liquid chromatography tandem mass spectrometry (LC-MS-MS) in positive ionization mode. The recoveries obtained (with mean values between 85 and 115% (obtained at different fortification levels) with RSD values below 10% in most cases, confirm the usefulness of the proposed methodology for the analyses of these kind of complex samples with a high fat content. Moreover, the obtained detection limits, which were below 5 microg kg(-1) by LC-MS analyses and ranged from 10 to 60 microg kg(-1) by GC-MS meet the requirements established by the olive oil pesticide regulatory programs. The method was satisfactorily applied to different olives and olive oil samples. PMID- 15830945 TI - Controlled oxidative protein refolding using an ion-exchange column. AB - Column-based refolding of complex and highly disulfide-bonded proteins simplifies protein renaturation at both preparative and process scale by integrating and automating a number of operations commonly used in dilution refolding. Bovine serum albumin (BSA) was used as a model protein for refolding and oxido-shuffling on an ion-exchange column to give a refolding yield of 55% after 40 h incubation. Successful on-column refolding was conducted at protein concentrations of up to 10 mg/ml and refolded protein, purified from misfolded forms, was eluted directly from the column at a concentration of 3 mg/ml. This technique integrates the dithiothreitol removal, refolding, concentration and purification steps, achieving a high level of process simplification and automation, and a significant saving in reagent costs when scaled. Importantly, the current result suggests that it is possible to controllably refold disulfide-bonded proteins using common and inexpensive matrices, and that it is not always necessary to control protein-surface interactions using affinity tags and expensive chromatographic matrices. Moreover, it is possible to strictly control the oxidative refolding environment once denatured protein is bound to the ion exchange column, thus allowing precisely controlled oxido-shuffling. PMID- 15830946 TI - Separation of atropisomeric 1,4,5,6-tetrahydropyrimidinium salts by chiral HPLC and determination of their enantiomerization barriers. AB - Chiral HPLC separation of a series of novel atropisomeric quaternary (1) and ternary (2) 1,2-disubstituted 1,4,5,6-tetrahydropyrimidinium salts bearing disymmetric aryl groups in positions 1 and/or 2 is described. A screening of different polysaccharide stationary phases (OD-R, OJ-R and AD-RH) and chromatographic conditions allowed for partial or baseline resolution of 16 over 26 compounds. When a semi-preparative separation was achieved, the corresponding enantiomerization barriers were determined employing the off-column method. The experimental data were compared inter se in order to establish the factors influencing the magnitude of the barriers and with those corresponding to the parent amidines. PMID- 15830947 TI - Determination of L-carnitine in food supplement formulations using ion-pair chromatography with indirect conductimetric detection. AB - A novel method for the determination of L-carnitine in food supplement formulations was developed and validated, using ion-pair chromatography with indirect conductimetric detection. The chromatographic method was based on a non polar (C18) column and an aqueous octanesulfonate (0.64 mM) eluent, acidified with trifluoroacetic acid (5.2 mM). The retention time was 5.4 min and the asymmetry factor 0.65. A linear calibration curve from 10 to 1000 microg/ml (r= 0.99998), with a detection limit of 2.7 microg/ml (25 microl injection volume), a repeatability %RSD of 0.8 (40 microg/ml, n = 5) and reproducibility %RSD of 2.6 were achieved. The proposed method was applied for the determination of carnitine in oral solutions and capsules. No interference from excipients was found and the only pretreatment step required was the appropriate dilution with the mobile phase. Recovery from spiked samples was ranged from 97.7 to 99.7% with a precision (%RSD, n = 3) of 0.01-2.1%. PMID- 15830948 TI - Characterization of phenyl-type HPLC adsorbents. AB - A set of different phenyl-modified HPLC adsorbents were characterized in terms of their surface area, pore volume, and bonded phase volume using low temperature nitrogen adsorption (LTNA). Adsorbents pore volume and interparticle volume were also measured using HPLC. Comparison of the pore volumes assessed with LTNA and HPLC suggests a compact molecular arrangement for all bonded phases studied. Simple and effective method for determination of the exact mass of adsorbent and total surface area in the column is suggested. PMID- 15830949 TI - Effect of anionic additive type on ion pair formation constants of basic pharmaceuticals. AB - Due to their beneficial effect on selectivity, peak shape, and sample loading, the use of mobile phase anionic additives, such as formate (HCOO-), chloride (Cl ), and trifluoroacetate (CF3COO-), is increasing in both reversed-phase chromatography (RPLC) and liquid chromatography-mass spectrometry (LC/MS). Similarly, perchlorate is a common "ion pair" agent in reversed-phase separation of peptides. Although many studies have suggested that anions effect in chromatography is due to the formation of ion pairs in the mobile phase between the anions and cationic analytes, there has been no independent verification that ion pairs are, in fact, responsible for these observations. In order to understand the mechanisms by which anionic additives influence retention in chromatography and ionization efficiency in electrospray mass spectrometry, we studied the formation of ion pairs between a number of prototypical basic drugs and various additives by measuring the effect of anionic additives on the electrophoretic mobility of the probe drugs under solvent conditions commonly used in chromatography. For the first time, ion pair formation between basic drugs and anionic additives under conditions commonly used in reversed-phase liquid chromatography has been confirmed independently with all anions (i.e. hexafluorophosphate, perchlorate, trifluoroacetate, and chloride) used in this study. We measured ion pair formation constants (Kip) for different anionic additives using capillary electrophoresis (CE) and obtained quantitative estimates for the extent of ion pairing in buffered acetonitrile-water. The data clearly indicate that different anionic additives ion pair with cationic drugs to quite different extents. The ion pair formation constants show a clear trend with the order being: PF6- > ClO4- > CF3COO- > Cl-. However, the extent of ion pairing is not large. At a typical RPLC mobile phase additive concentration of 20mM, the percentages of the analytes that are present as ion pairs are about 15%, 6%, and 3% for hexafluorophosphate, perchlorate, and trifluoroacetate, respectively. The fraction of the analytes present as a chloride pair is even smaller. PMID- 15830950 TI - Determination of aromatic hydrocarbons in asphalt release agents using headspace solid-phase microextraction and gas chromatography-mass spectrometry. AB - The possibility of quantitative analysis of aromatic hydrocarbons in oil-based asphalt release agents was investigated using headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC MS). The target analytes studied were benzene, toluene, ethylbenzene, p-, m-, and o-xylene (BTEX) and 1,3,5-trimethylbenzene and 1,2,4-trimethylbenzene. Experimental parameters influencing HS-SPME efficiency were studied (equilibration time between sample and headspace and between headspace and SPME fiber, sample amount and sample matrice effects). A HS-SPME method using hexadecane as a surrogate matrice was developed. The detection limit was estimated as 0.03-0.08 ppm (w/w) for the target analytes investigated. Good linearity was observed (R2 > 0.999) for all calibration curves at high, medium and low concentration level. The repeatability of the method (RSD, relative standard deviation) was found to be less than 10% (generally less than 5%) in triplicate samples and approximately 2% at eight consecutive tests on one and the same sample. The accuracy of the method given by recovery of spiked samples was between 85 and 106% (generally between 95 and 105%). The HS-SPME method developed was applied to four commercially available asphalt release agents. External calibration and standard addition approaches were investigated regarding accuracy. The results showed that standard addition generates higher accuracy than external calibration. The contents of target aromatic hydrocarbons in the asphalt release agents studied varied greatly from approximately 0.1-700 ppm. The method described looks promising, and could be a valuable tool for determination of aromatic hydrocarbons in different types of organic matrices. PMID- 15830951 TI - Application of purge and trap extraction and gas chromatography for determination of minor esters in cider. AB - The validation of a method based on the purge and trap technique combined with gas chromatography-mass spectrometry-flame ionization detection has been carried out in order to apply it to the analysis of ciders. Although 49 compounds were identified, our work was focused on the study of nine minor esters, obtaining recoveries ranging between 93% for ethyl decanoate and 117% for ethyl 3 methylbutyrate, and a precision (RSDs) ranging between 2.2% for hexyl acetate and ethyl decanoate and 10.9% for isopentyl acetate. To demonstrate the feasibility of the procedure, the method was applied to the analysis of commercial ciders. PMID- 15830952 TI - Specific differentiation in Vicia genus by means of capillary electrophoresis. AB - Capillary electrophoresis has been applied to the discrimination of 13 Vicia species, belonging to four sections of Vicia genus. The studied species necessitate of plant growing tests or DNA molecular markers to be distinguished being their seeds morphologically very similar. Alcoholic/saline extracts from dry cotyledons were separated in uncoated fused-silica capillary with iminodiacetic acid isolectric buffer containing hydroxypropylmethylcellulose and acetonitrile. The low intra-specific variation observed for 11 species, suggests that this approach is suitable to carry out species discrimination. Species specific peaks were identified for V. articulata, V. atropurpurea, V. bithynica, V. benghalensis, V. disperma, V. ervilia, V. monantha, V. sativa and V. villosa. Conversely, V. lutea, V. melanops and V. peregrina, showing very similar electrophoregrams, require other methodological approaches to be discriminated. The discussed CE method appears to have a potential to be regarded as an alternative tool to identify some Vicia species being far less expensive and time consuming than plant growing tests and DNA molecular markers. PMID- 15830953 TI - Capillary zone electrophoretic studies of ion association between inorganic anions and tetraalkylammonium ions in aqueous-dioxane media. AB - Ion association between inorganic anions and symmetrical tetraalkylammonium ions, R4N+ (R = Me, Et, Pr, n-Bu, n-Am, and 2-methyl butyl {isoamyl = iAm}) was investigated using ordinary silica capillary by capillary zone electrophoresis. An improved version of the Williams-Vigh method was used for the first time to measure the mobilities of the inorganic anions. Plots of log Kass against log dielectric constant in various media, revealed a smaller change in Kass compared to dielectric constant. These plots suggest that the Bjerrum's equation is inadequate in accounting for the associations of ions in a CZE setup. PMID- 15830954 TI - Concentration of polyaromatic hydrocarbons in water to sodium dodecyl sulfate gamma-alumina admicelle. AB - Polyaromatic hydrocarbons (PAHs) in water were concentrated into sodium dodecyl sulfate (SDS)-gamma-alumina and di-2-ethylhexyl sodium sulfosuccinate (Aerosol OT, AOT)-gamma-alumina admicelles. The comparison of the binding constants (Kad[={adsorbed concentration of the solute (mol/g surfactant)}/{the concentration in the bulk aqueous phase (mol/ml)}] indicated almost the same extraction abilities of the both admicelles. However, better and more reproducible recovery was obtained in the concentration of PAHs into the SDS gamma-alumina admicelle. PAHs in tobacco smoke that were trapped in water were successfully concentrated into SDS-gamma-alumina admicelle for the HPLC analysis. PMID- 15830955 TI - Focused microwave aqueous extraction of chlorophenols from solid matrices and their analysis by chromatographic techniques. AB - Open-vessel focused microwave (FMW) extraction with a purely aqueous carbonate solution was used for the extraction of chlorophenols from various solid matrices. After SPE on C18-bonded silica, the analytes were determined as such by LC-UV or, as their acetyl derivatives, by GC-ECD. The FMW aqueous extraction is efficient and rapid and no organic solvents are used. PCP was detected in several solid samples, with recoveries of 101-115% (RSD, 2-4%) relative to Soxhlet extraction. Similar recoveries were obtained for the other chlorophenols for spiked samples. PMID- 15830956 TI - Analysis of herbicides in water using temperature-responsive chromatography and an aqueous mobile phase. AB - A simple and rapid method has been developed for herbicides in water using temperature-responsive liquid chromatography (LC) and a column packed with poly(N isopropylacrylamide) (PNIPAAm), a polymer anchored on the stationary-phase surface of modified silica. PNIPAAm reversibly changes its hydrophilic/hydrophobic properties in water in response to temperature. The method was used to determine five sulfonylurea and three urea herbicides. Separation was achieved with a 10 mM ammonium acetate (pH 3.0) isocratic aqueous mobile phase, and by changing the column temperature. The analytes were extracted from water by off-line solid-phase extraction (SPE) with an N-vinyl-pyrrolidone polymer cartridge. The average recoveries of the eight herbicides from spiked pure water, tap water and river water were 70-130% with relative standard deviations (RSDs) of <10%. The limits of quantitation (LOQ) of the eight herbicides were between 1 and 4 microg l(-1). PMID- 15830957 TI - Neurolysis: is it beneficial or harmful? AB - The term internal neurolysis means removal of fibrotic tissue inside a nerve trunk. Unfortunately the term was used for procedures with complete isolation of fascicles with all consequences like damage of links between the fascicle and impairment of blood supply. The conclusion based on some negative experiences that all surgery within a nerve trunk has to be avoided cannot be accepted. Neurolysis within a nerve trunk, id est within the epineurium, is a step-wise procedure to decompress fascicles from a constricting fibrosis. It stops immediately if this aim is achieved or continues with resection and reconstruction if an irreparable damage is present. It is better to use terms that describe exactly what was done and abandon the ill-defined term "internal neurolysis". Fibrosis of the paraneurium remains outside the epineurium but causes the same consequences as fibrosis of the epineurium. PMID- 15830958 TI - TOS pathophysiology and clinical features. AB - The authors present 280 patients operated on for thoracic outlet syndrome (TOS). In a first group of patients anatomical variants were the striking findings. The underlying factor for TOS development is therefore a well defined structural condition and its pathogenetic mechanism is known to be a nerve fibre compression. In a second group there was no specific salient finding but a postural deviation. The unique pathological features were adhesions of the brachial plexus to the scalenus muscle. Consequently its pathogenetic mechanism is generally recognized as nerve fibre distraction. In all patients neurological, vascular and myofascial pain symptoms were observed before the operation. Neurological and vascular pain disappeared after surgery, while the myofascial pain remained. The authors believe that especially in the second, larger group of patients enhancement of the pain-immobility-fibrosis loop is the central pathogenetic factor on which surgical therapy is successful, and that myofascial hemisyndrome--probably arising from a long-standing postural deviation--is not a TOS dependent symptom. In TOS, therefore, there is a pain loop that cannot be resolved by surgical therapy alone. The connection between myofascial pain syndrome and TOS might explain the many controversial opinions regarding frequency, results and surgical possibilities of this lesion. PMID- 15830959 TI - Post-traumatic thoracic outlet syndrome (TOS). AB - TOS is a compressive non-tumorous syndrome of the brachial plexus. It is possible, however, to consider as TOS the irritative and lesional plexus syndrome following trauma as long as compression (or traction) on the nerves is triggered by long-lasting pathological changes of the area after trauma. Overload work of judges and lawyers after traffic accidents does not help to remind the real victim's problem, that is stretching of the neck soft tissues during head acceleration-extension. This movement is due to a forward acceleration. Both the car and the victim's trunk are violently pushed forward while the head does not move fast enough so that it is actually pushed backwards. The mandibula is even slower than the head and this leads to an opening of the mouth with possible temporomandibular joint (TMJ) dislocation. If there is nothing stopping the neck extension, like an appropriate headrest, the momentum is only resisted by cervical soft tissue stretching. Prolonged antalgic contracture and motor neglect may contribute to connective tissue changes and development of microadherences. Final result is fibrosis of paraneurium. The pain-immobility-fibrosis loop is of basic importance in the development of this syndrome. PMID- 15830960 TI - Brachial plexus injuries: regeneration timing and prognosis in patients without need for urgent operation. Preliminary results on truncus primarius superior. AB - BACKGROUND: In Italy, and more generally in the industrialised countries, traumatic nerve lesions have become more frequent. It is commonly accepted that it is necessary to wait 6 months after injury to suggest surgery if movement does not appear. In the scientific literature, there is no systematic clinical evidence of nerve regeneration timing after trauma., especially regarding brachial plexus. METHOD: We have performed a follow-up study of 15 consecutive patients with traumatic brachial plexus injuries involving truncus primaries superior without need for urgent surgery. In each patient an extensive clinical and neurophysiological evaluation was performed to detect the kind of lesion, level of lesion, severity of lesion and the outcome. FINDINGS: In our sample, some cases improved within a few weeks. This rapid improvement may be attributed to resolution of neuroapraxic block, in other cases slower improvement occurred due to rearrangement of motor units and axonal regeneration. In some cases voluntary activity clinically appeared after more than 6 months following injury. CONCLUSIONS: The current study is preliminary, but it provides evidence that a period of 6 months may not be sufficient for the reappearance of clinical movement. Moreover, current results confirm that neurophysiological evaluation may be a highly prognostic tool in traumatic nerve lesions. We hope that our study together with other data may provide us a timetable for expected nerve regeneration. PMID- 15830961 TI - Thoracic outlet syndrome due to hyperextension-hyperflexion cervical injury. AB - Posttraumatic brachial plexus entrapment in fibrotic scarring tissue is taken into consideration as the cause of complaints for patients who suffered a hyperextension-hyperflexion cervical injury. All 54 patients included in this analysis where symptom-free before the accident and subsequently complained for pain, paresthesia and slight weakness in the arm. In 14 neurological signs of brachial plexus entrapment were observed. Electroneurophysiological, summary index testing was positive for a brachial plexus involvement in all cases. Conservative measures, comprising physical therapy and vasoactive drugs were applied for a period of 6 to 12 (mean 8.4) months; surgical procedure of neurolysis was then proposed in 39 cases to solve the problem. Thirty-two patients were operated on. Twenty of these had a neat improvement on a 6-month to 1-year follow-up. Seven patients had refused surgery; of these 6 patients had clinical worsening at the same follow-up period while 1 remained unchanged. All patients with clinical symptoms not reversed after some time post-injury should be investigated for a possible brachial plexus entrapment. PMID- 15830962 TI - Whiplash injury. TOS and double crush syndrome. Forensic medical aspects. AB - In this article the author wants to specify that the whiplash syndrome is underestimated, even by the specialists. In particular the complications aren't taken into correct consideration, above all if they concern the brachial plexus, especially regarding the TOS syndrome and double-crush syndrome. This is a problem also among the experts who have to make an evaluation in the field of insurance. PMID- 15830963 TI - Vascular Thoracic Outlet Syndrome staging and treatment. AB - Thoracic Outlet Syndrome (TOS) is a well known lesion. Sophisticated imaging techniques can clearly highlight any anatomical damage and a wide range of therapeutic choices are available. It would seem obvious that any given patient should obtain the same treatment irrespective of the medical institution he contacts, but this is not the case. Instead each specialist may recommend different treatments: physiatrist, neurologist, surgeons (thoracic, vascular, neuro, orthopedic). Everyone preserves his specific language and there is no univocal treatment plan consensus for this complex syndrome. Evidently, the correct staging of TOS is still an unresolved question. In order to solve this problem, we collected all clinical and instrumental aspects of the syndrome into a clear, precise classification. Similar to TNM staging of malignant diseases, we used a grouping model based on the three mainly involved anatomical structures: N (= Nerves; brachial plexus and sympathetic fibers), A (= Artery; subclavian axillary), V (= Vein; subclavian-axillary). We named it the NAV staging of TOS. A retrospective examination of our case records confirmed a valid and useful correlation between the proposed NAV staging and the therapeutic procedures that were actually applied. It is now essential to perform a multi-centre study to extend the validity of our staging. PMID- 15830964 TI - Suprascapular nerve entrapment. AB - It is important to be aware of neuropathy involving the suprascapular nerve. While direct trauma to the suprascapular nerve is the usual cause (direct blow to the base of the neck or posterior shoulder, shoulder dislocation or fracture), the problem may result from overuse injuries (such as repetitive tennis serving or spiking of a volley ball), excessive horizontal adduction, weight lifting, backpacking or no apparent reason. These last three years we have operated 8 cases of suprascapular nerve neurolysis at the level of suprascapular incision, and section of the transverse scapular ligament through the back supraspinal approach. PMID- 15830965 TI - Multicenter study on carpal tunnel syndrome and pregnancy incidence and natural course. AB - OBJECTIVE: To evaluate the incidence of carpal tunnel syndrome (CTS) in pregnancy through a validated and multiperspective assessment of CTS and to assess the course of carpal tunnel syndrome after pregnancy. METHODS: During 2000-2001, the Italian CTS study group in 7 Italian centers studied the occurrence of CTS in women during the last period of pregnancy. The group enrolled and followed-up (10 15 months) 63 women during and after pregnancy with multiple measurements of CTS. In addition to the physician-centered and neurophysiologic traditional evaluations, a validated patient-oriented measurement to obtain more comprehensive and consistent data for severity of symptoms and functional impairment was adopted. RESULTS: CTS was clinically diagnosed in more than half of women (62%). Neurophysiological evaluation provided diagnosis of CTS in around half of women (43% were positive in one hand at least). Comparison of baseline and follow-up data showed a significant spontaneous improvement of patient oriented and neurophysiologic measurements. Nevertheless, about half of women with CTS during pregnancy still complained of CTS symptoms one year after delivery. CONCLUSIONS: Our observations confirmed the frequent occurrence of CTS in pregnancy. At follow-up we observed that most CTS cases improve spontaneously without treatment but only in half of women CTS symptoms disappeared one year after delivery. PMID- 15830966 TI - Endoscopic carpal tunnel release surgery: retrospective study of 390 consecutive cases. AB - Endoscopic carpal tunnel release (ECTR) surgery was developed by Okutsu and Chow in 1989. Many reports indicated that the endoscopic technique reduces postoperative morbidity with minimal incision, minimal pain and scarring, a shortened recovery period and high level of patient satisfaction. To evaluate these reports, a retrospective study was conducted with 390 procedures of two portal Chow technique for idiopathic carpal tunnel syndrome. Follow-up was performed at 1, 3 and 6 months and overall results were backed up by telephone questionnaire (Health Outcomes Carpal Tunnel Questionnaire, Health Outcomes, Bloomington, MN, USA). Results were favourable in 98% and 2% unfavorable for persistent pain. Rate of satisfaction of the patients was 90%. Average time of patient's return to work was 20 days. Eleven procedures (2.8%) were converted to open release. There was one case (0.2%) of incompleted section of the perineurium due to failure of endoscopic visualization of the ligament. In this case the procedure was converted to open and was completed with perineurium sutura. In six cases (1.5%) there were injury to superficial palmar arch. During the follow-up period there were no recurrences and no re-exploration. The mean preoperatively obtainable distal motor latency (DML) and sensory conduction velocity (SCV) values were 6.7 m/s and 29.2 m/s, respectively. The mean DML and SVC values at final follow-up were 3.8 msec and 42.3 m/s, respectively. In conclusion, ECTR can be used in the carpal tunnel syndrome and is a reliable alternative to the open procedure with excellent self-report of patient satisfaction. Reduced recovery period with minimal tissue violation and incisional pain can be expected. PMID- 15830967 TI - Wrist median nerve motor conduction after end range repeated flexion and extension passive movements in Carpal Tunnel Syndrome. Pilot study. AB - Carpal Tunnel Syndrome (CTS) can be due to a variety of different pathological conditions. These etiological and epidemiological differences may explain the non homogeneous response to ordinary conservative therapeutical options observed in this syndrome. The aim of our study was to investigate on the possibility of identifying different sub-groups of patients among conservatively treatable CTS with different susceptibility to physiotherapeutic treatments. We decided to utilize an objective approach measuring some median motor nerve function parameters. Short term variations of Compound Motor Action Potential (CMAP) from the thenar eminence were compared in two groups of 55 hands (CTS patients and normal controls) after performance of two different types of end range passive movement. We found a different distribution of CMAP amplitude modifications within a sub-group of patients that suddenly improved more than the controls after two series of 10 end range passive flexions or after two series of ten end range passive extensions. Amplitude changes proved to be much more useful than latency variation studies in the provocative test neurophysiological approach. The method we propose appears to be useful for better surgical indication and/or for improvement of conservative therapeutic choice. PMID- 15830968 TI - Microsurgical treatment of lumbosacral plexus injuries. AB - Surgical treatment of lumbar and sacral plexus lesions is very rarely reported in the literature. The incidence of the involvement of these nervous structures in traumatic lesions of different etiology is probably much higher than believed, and surgical treatment should be taken into consideration more often. In this paper the experience derived from the surgical treatment of 15 cases is reported. Different surgical approaches have been employed according to ethiology, to level of nerve lesion and concomitant lesions of other organs. Patients who suffered a lesion in the lumbar or sacral plexus may have a very severe problem with deambulation since the leg may not be stable or may be unable to withstand the weight of the body. Pain syndrome in these patients may be a very severe obstacle to rehabilitation programs and to deambulation and everyday activity. Microsurgical nerve treatment in the retroperitoneal space is demanding both for the surgeon and for the patient but neurolysis and grafting procedures are possible also in this area. The resulting improvement of motor performance and the relief of pain are strong arguments in favor of this choice. Muscles benefitting most from surgery are the gluteal and femural muscles; more distant muscles, and particularly the anterior tibial nerve dependent muscles will gain minimal benefit from surgery. The relief from pain is relevant in all cases. PMID- 15830969 TI - Femoral nerve entrapment. AB - We present 30 cases of femoral nerve entrapment (1999-2003, age range 35-65 yrs), in 13 patients with diagnosis of idiopathic compression and 7 patients of neurovascular conflict. The compression, in the other 10 patients, was iatrogenic: 3 patients following cardiac catheterization for balloon valvotomy, 2 patients following intra-abdominal vascular surgery and 5 patients following laparoscopic hernia treatment. Microsurgical nerve decompression, and the elimination of neurovascular conflict gave satisfactory results. The best result has been observed in neurovascular conflict cases. PMID- 15830970 TI - Italian multicentre study of peroneal mononeuropathy at the fibular head: study design and preliminary results. AB - BACKGROUND: The most common entrapment in the lower extremity is peroneal mononeuropathy (PM) at the fibular head. Several studies of this condition have been published but, until now, no wide multicenter clinical-neurophysiological studies on PM are available. In recent years, multicenter studies have been suggested; moreover it is commonly accepted that a multiperspective approach provides more comprehensive results. METHOD: The Italian CTS and other entrapments Study Group has designed a strict clinical and neurophysiological protocol to carry out a wide multicentre study on PM at the fibular head. In addition to traditional clinical-neurophysiological evaluation, the group has also adopted validated disability and patient-oriented measurements in order to obtain more comprehensive and reliable data about this entrapment. The study was designed: 1) to identify predisposing factors; 2) to better assess the clinical picture; 3) to evaluate relationships between etiological, clinical and neurophysiological findings; 4) to evaluate the natural evolution of the entrapment. Study design is described. FINDINGS: During the period from November 2002 to January 2004, 69 patients were enrolled consecutively in eleven Italian centres. Our preliminary data show that PM involves men more frequently than women (M:F = 3.9:1). With regard to the predisposing factors, PM is idiopathic (16%) or due to surgery (21.7%), prolonged posture (23.2%), weight loss (14.5%), external compression (5.8%), arthrogenic cyst at the fibula (1.4%), trauma (10.1%); it also occurred in bedridden patients (7.3%). Unexpectedly, peroneal nerve lesions were due not only to surgical operation close to the peroneal region, but were also associated with thoracic-abdominal surgery. Usually PM involves both terminal branches; patients complain of motor deficit in 99.5% of cases, sensory symptoms in 87.9% and pain in 19.7%. CONCLUSIONS: Our preliminary results provide some interesting information and confirm the usefulness of multicentre and multiperspective studies to standardise the approach to nerve entrapment. PMID- 15830971 TI - Entrapment of crural branches of the common peroneal nerve. AB - Failed back surgery syndrome (FBSS) occurs in 30% of operated patients and represents a heavy problem both regarding disability and costs in first world countries. Among FBSS we found the possibility of a double crush syndrome: a disco-radicular conflict and a peripheral nerve entrapment. The latter, disguised by root compression symptoms, becomes evident only after spinal surgery. Clinical features are the same as for the restless leg syndrome. We found peroneal nerve crural branches entrapped where they crossed the fascia to reach the subcutaneous layer. Venous stasis during immobility caused presentation of symptoms. Neurolysis was performed, all cases were successful. Most of the patients were found to have myofascial pain syndrome (MPS). MPS patients "feel" entrapments more frequently than others not because of their specific pain tolerance but because they are more prone to develop them. PMID- 15830972 TI - Percutaneous cervical nucleoplasty using coblation technology. Clinical results in fifty consecutive cases. AB - Conventional open cervical discectomy, with or without bony fusion, in common neurosurgical knowledge is considered the standard treatment for cervical disc herniation. Percutaneous procedures are minimally invasive and offer decreased morbidity, require no bone graft and promise shorter recuperation time. Nevertheless, candidates for a percutaneous procedure as inclusion criteria must complain of symptoms related to contained herniated disc or focal protrusion. It does not substitute conventional open procedures required for extruded discs. We used the coblation technology for nucleoplasty of the cervical intervertebral discs. Early and long-term effects and/or complications observed with this procedure have not been reported yet. Fifty consecutive patients presenting with contained herniated cervical disc or focal protrusion causing compression of the cervical roots or cervical pain underwent a nucleoplasty procedure on the pathological disc. A randomized control group of twenty patients was treated conservatively with medical and physical therapy in the same period and completed the identical follow-up form. In the nucleoplasty group results were complete resolution of symptoms in 80% of cases, only 10% referred some residual cervical or radicular pain and are still under follow-up with a wait-and-see prospective. Patients who did not have a clinical resolution were treated with alternative traditional methods (10%). Despite the relative low cases number and the limited follow-up the encouraging results induce us to utilize this technique in well selected cases. PMID- 15830973 TI - Intradiscal injection of oxygen-ozone gas mixture for the treatment of cervical disc herniations. AB - For disc herniations the use of open surgical approaches is reduced since new percutaneous methods allowing shrinkage of the disc and improvement of the radicular function are gaining interest. Studies on the spontaneous disappearance of disc fragments have demonstrated autoimmune responses with a chronic inflammatory reaction. Also radicular pain has been shown to be mostly due to biochemical mechanisms. Researchers in different fields surprisingly noticed that a brief, calculated, oxidative stress by ozone administration may correct a persistent imbalance due to excessive, chronic oxidative injury. Oxygen-ozone gas injection in painful patients has a dramatic effect on clinical symptoms. On these bases the intradiscal injection of oxygen-ozone gas has been conceived. We report the treatment on a series of patients affected by cervical disc pathology, treated by intradiscal injection of oxygen-ozone gas mixture. The effects both on pain and on radicular dysfunction are impressive. The morphological effect of the treatment was also evaluated by pathological examination. PMID- 15830974 TI - Percutaneous nucleoplasty for discoradicular conflict. AB - Minimally invasive techniques for the treatment of degenerative pathology of the spine have come to be preferred by surgeons since the destructive effect on bony structures is eliminated and scar formation is dramatically reduced. A critical review of the pathogenetic mechanisms for low back pain and sciatalgia has recently yielded that mechanical compression is one but non essential component of the matter. The importance of chemical irritative processes is stressed. Coblation nucleoplasty is one of these minimally invasive techniques. It provokes ablation of the nucleus of the disk by a controlled thermal effect produced by radiofrequency. By this procedure one to two ml of tissue are colliquated in a few minutes. From February 2001 to May 2003 we treated 1390 patients for of lumbosciatalgic pain caused by disc pathology. The alteration consisted of disc bulging or contained disc herniation. Exclusion criteria as provided by the protocol of the multicentric study conceived by Conor O'Neill have been respected. This technique has been conceived in order to obtain progressive results in cases of contained disc herniation which has scanty natural tendency to shrinkage, as demonstrated by several studies on the natural history of evolution of this pathology. Contained disc herniation is a pathology most difficult to manage by conservative procedures, physiotherapy and drugs, but we all agree that open surgery should be avoided. By this minimally invasive procedure the patient will not be compelled to abandon physiotherapy and his normal daily activities for more than a few days. PMID- 15830975 TI - CT-guided oxygen-ozone treatment for first degree spondylolisthesis and spondylolysis. AB - Aim of this study was to assess the therapeutic outcome of CT-guided periganglionic infiltration of oxygen-ozone and injection of the gas mixture into the lysis points in patients with first grade spondylolisthesis and spondylolysis. We selected 18 patients presenting with low back pain and sciatica resistant to physical and medical management with a radiological diagnosis of spondylolisthesis and spondylolysis subsequently confirmed on CT scan. Following CT-guided bilateral periganglionic O2-O3 infiltration and injection into the lysis points, 15 patients (83.3%) obtained a complete remission of pain. None of the patients reported pain recurrence at clinical follow-up visits one, three and six months after treatment. Oxygen-ozone therapy administered in this way is even more effective than CT-guided periganglionic infiltration alone as it has an additional anti-inflammatory and analgesic effect on the nerve structures in the neural arch, namely Luschka's recurrent nerve. PMID- 15830976 TI - Ozone chemonucleolysis in non-contained lumbar disc herniations: a pilot study with 12 months follow-up. AB - STUDY DESIGN: Prospective case series with six and twelve months follow up. OBJECTIVE: To observe clinical and morphological results of the intradiscal ozone chemionucleolysis in patients affected by non-contained lumbar disc herniations. METHODS: 30 patients were included in the study on the base of precise inclusion and exclusion criteria. The patients were followed on 6 and 12 months period by Visual Analogic Scale (VAS), Roland Morris Disability Questionnaire (RMDQ) and Overall Patient Rating Scale (OPRS). Disc herniation volume morphology was evaluated at 5 months by control MRI scanning. RESULTS: Twenty-seven patients (90%) showed a statistically significant improvement in pain (P < 0.001, Wilcoxon test) and function (P < 0.001, Wilcoxon test), on VAS and RMDQ evaluation, respectively. The mean satisfaction with the treatment on OPSR was 79.3%, with 24 patients referring satisfaction equal or greater than 80%. There were no major complications related to the procedure. CONCLUSIONS: The results of this study indicate the ozone chemonucleolysis as a possibly effective modality of treatment in patients affected by signs and symptoms of non-contained lumbar disc herniations that have overpassed conservative measures and have not yet fulfilled the indications for open surgical treatment. PMID- 15830977 TI - Micro-endoscopic-discectomy (MED) for far lateral disc herniation in the lumbar spine. Technical note. AB - This study describes a new experience of the authors in the treatment of extraforaminal disc herniation via the micro-endoscopic far lateral approach to establish a less traumatic approach to extraforaminal disc herniation with less stay in hospital and less cost. Seventeen patients who underwent surgery for extraforaminal disc herniation were analysed and long-term follow up was done revisiting all of them in hospital. The results of surgical decompression via the micro-endoscopic far lateral approach were good in all patients with minimal discomfort. There was complete resolution of leg pain presented. Dysesthesia subsided after 2-3 weeks. Extraforaminal disc herniation can be diagnosed with the aid of CT scan and MRI. The minimally invasive surgical treatment via the micro-endoscopic far lateral approach, in our initial experience, has a high rate of succes. PMID- 15830978 TI - A long-term review of 50 patients out of 506 treated with automated percutaneous nucleotomy according to Onik for lumbar-sacral disc herniation. AB - At the Orthopaedics and Traumatology Unit of the Palmanova Hospital, between 27 October 1989 and 31 December 2003 we performed 506 automated percutaneous nucleotomies according to Onik for the treatment of lumbar disc hernia. The survey of 50 reviewed cases after evaluation of the subjective and objective clinical pictures according to the Cabot method allowed us to come to the conclusion that this percutaneous methodology is suitable to relieve damaged discs from compression. It is also well accepted by patients because it is not too traumatic, it requires short-term hospitalisation, presents no risk of post operative fibrosis and does not create complications for the eventual traditional operation when unsuccessful. It is extremely important to accurately select the candidates keeping in mind the original indications given by Onik for percutaneous discectomy for which--in case of contained disc herniation--leg pain (sciatalgia) is more severe than low back pain affecting the lumbar region. PMID- 15830979 TI - Surgical intradiscal decompression without annulotomy in lumbar disc herniation using a coblation device: preliminary results. AB - Annulotomy is a mandatory step to perform intradiscal decompression to resolve a disco radicular conflict. However, this manoeuvre can lead to post surgical complications such as vertebral instability and back pain. Coblation assisted microdiscectomy (CAM procedure) allows a quoted removal of disc without anulus damage. PMID- 15830980 TI - CAM versus nucleoplasty. AB - In recent years the general trend in spinal surgery has been reduction and minimalization. In general, all these have shown a moderate or good clinical result but they have been associated with serious sequelae. Plasma-mediated electrosurgery, widely used in other medical fields, has demonstrated to be well suited for this new indication. To perform the Nucleoplasty (Coblation) and the CAM (Coblation-Assisted Microdiscectomy) is use the Perc-DLE SpineWand connected to a System 2000 generator (ArthoCare Corp., Sunnyvale, CA) was used. The device functions via plasma-mediated electrosurgery (Coblation) and differs from traditional electrosurgery. From a small sample 64 operated patients with contained disc herniation were analysed and classified into those who underwent percutaneous disc decompression (PDD) using coblation technology and patients who underwent CAM. All patients who presented with PDD were considered candidates for open surgery but all of them opted for the new technique. There was no contraindication. They had discogenic low back pain and/or leg pain and the procedure was performed on an outpatient basis. Follow-up data was of 1 to 12 months. Patients' gender distribution for PDD was 65% (41.6) male, 35% (22.4) female with a mean age of 43 years. The average duration of pain before nucleoplasty was of 18 months and none of them had previous lumbar surgery. At 6 to 12 months, 80% of the patients demonstrated an improvement in pain scores (75% very good, 5% good, 15% improved but not good, and 5% no effect). None of the patients was worse. Results indicate that Nucleoplasty may be an efficacious minimally invasive technique for the treatment of symptoms associated with contained herniated disc. However, randomized controlled studies are required to know with more precision the role of this procedure. CAM procedure (13 cases) is an excellent method in cases of root compression that needs liberation or in spine stenosis. PMID- 15830981 TI - Paraspinal approach to the far lateral disc herniations: retrospective study on 42 cases. AB - Forty-two patients underwent surgery for far-lateral disc herniations. Average patient age was 45.1 years, 28 patients were male and 14 female. The level concerned most was L4-5 disc (55%). A paramedian muscle-splitting intertransverse approach is described for this type of disc herniation. This method allows direct visualization of the disc and root involved and does not provide bone resection and important surgical manipulation. Ninety-one percent of the patients reported excellent or good results according to Macnab outcome criteria with an average follow-up of 32.5 months (range 1-5 years). Twelve patients (28.6%) developed postoperative transient radicular pain that disappeared in 15-21 days after analgesic therapy. There were no recurrences or spinal instability. The paramedian muscle-splitting intertransverse approach is a rational technique. Its advantages are that the spine is not opened and spinal stability is maintained. It requires minimal soft-tissue and bone resection and the herniated disc is directly visualized. Moreover, it contains minimal manipulation of the neuro vascular structures and avoids significant muscle retraction. However, it requires an adequate learning curve and good familiarity with microsurgical techniques. PMID- 15830982 TI - Surgical radio-frequency epiduroscopy technique (R-ResAblator) and FBSS treatment: preliminary evaluations. AB - Failed back surgery syndrome represents a heterogeneous situation that suggests a fibrosis or neuroinflammatory genesis. The social cost related to this issue are enormous. Several surgical techniques have been applied to FBSS patients with controversial effectiveness. In 1998 we evaluated the efficiency and limits of epiduroscopy treatment; it proved to be effective in 75% of cases, but in 45% of cases it needed to be repeated after 12 months. Therefore we subjected 14 patients, who had previously experienced a short temporary benefit by using a traditional epiduroscopic approach, to a new epiduroscopy fibrolysis using a radio-frequency device named "R-Resablator Epiduroscopy". Clinical evaluation was performed before myeloscopy and after 1-3-6 months. After myeloscopy, 93% of patients reported a general improvement. Among the latter, pain was reduced by 90% in 8 patients, by 60-70% in 5, and by less than 30% in 1. CONCLUSION: It can be concluded that RF-Epiduroscopy offers greater therapeutic benefit than traditional epiduroscopy or other surgical techniques. Furthermore, RF Epiduroscopy is more easily performed and repeated. PMID- 15830983 TI - Disc coablation and epidural injection of steroids: a comparison of strategies in the treatment of mechanical spinal discogenic pain. AB - In this study two strategies in the treatment of Mechanical Spinal Discogenic Pain have been compared: Disc Coablation and Epidural Injection of Steroids. In 2003 50 patients treated with one or two epidural injections have been selected "ad random" and 50 patients treated with disc coablation. Comparison of the data indicated an improvement of average VAS when relaxed for both groups (p < 0.01), while after slight-moderate strain, this value was significant only after coablation (p < 0.001). Finally, average VAS was clearly lower (p < 0.01) after coablation as compared to epidural injections. PMID- 15830984 TI - Preliminary results of a soft novel lumbar intervertebral prothesis (DIAM) in the degenerative spinal pathology. AB - The authors report a series of 43 patients suffering from lower limb pain, almost constantly associated with chronic or acute backpain, treated by microsurgical nerve root decompression and by implantation of a soft intervertebral prothesis (DIAM). Satisfying results were obtained in 97% of cases, inducing the authors to consider the device a reliable tool for curing low-back pain and sciatica. Selection criteria are exposed and discussed. PMID- 15830985 TI - Lumbar spinal decompression with a pneumatic orthesis (Orthotrac): preliminary study. AB - AIM: We present a preliminary study on the conservative treatment of chronic low back pain (LBP) using an easy to manage and extremely practical orthesis. It consists of a pneumatic custom made lumbar vest (Orthotrac), which permits both support-stabilisation and decompression. This system is versatile since the patient is not impeded and can perform any activity while wearing it. MATERIAL: The study included 41 patients (23 males and 18 females, aged between 19 and 25 years) with radicular pain due to degenerative discopathy including: dark disc, discal protrusion with neural foramina involvement, stenosis of the foramina, syndrome of the facets, Grade 1 listhesis. Patients had to wear the Orthotrac vest according to a precise protocol, 60 minutes 3 times a day for 5 weeks. RESULTS: 32 patients (78%) have showed a significant subjective and clinical improvement with subsequent better quality of life. All patients referred a decrease or disappearance of radicular pain. Outcome measures were evaluated according to SF-36 system which is used in clinical practice and research. As in any innovative therapy, selection of patients is extremely important. The pneumatic vest is not indicated in all patients, but it can play an important role in non-surgical therapy for LBP. CONCLUSION: The system seems to give an effective spinal decompression and deserves a careful consideration when lumbar discal disease is treated conservatively. Further multicenter and interdisciplinary studies on a greater number of patients are obviously needed to confirm these preliminary results. PMID- 15830986 TI - The different outcomes of patients with disc herniation treated either by microdiscectomy, or by intradiscal ozone injection. AB - Disc herniation with radiculopathy and chronic discogenic pain are the result of degenerative processes. Treatment approach in face of this problem has largely been debated in the last years. A number of reviews on surgical treatments in the '80s and '90s have been published and various new techniques have been introduced among which ozone discolysis is one non-invasive intradiscal treatment method. In a 3-year follow-up period we have investigated the different outcomes of 150 patients who received microdiscectomy and 150 patients who received intradiscal ozone injection. In this series results are in favour of discolysis for contained disc herniations and of microdiscectomy for large migrated fragments with pain so severe that open surgery was obligatory. Apart from this, our results with the two techniques are equivalent also concerning mild neurological motor deficits. PMID- 15830987 TI - Quality of life, clinical and neurophysiological picture in patients operated on for lumbar stenosis. AB - BACKGROUND: In lumbar stenosis (LS) patients, clinical, neuroradiological and neurophysiological findings were not related to validated measurements of the outcomes that are more relevant to patients such as functional status and symptoms. METHOD: We have retrospectively studied 30 patients surgically treated for LS. We have evaluated the patients by means of self-administered questionnaires (SF-36), clinical examination, and neuroradiological and neurophysiological measurements and we have registered preoperative and follow-up clinical and neurophysiological findings. Finally we evaluated the relations between patient-oriented data and validated conventional clinical and neurophysiological measurements. FINDINGS: The comparison between pre- and post operative clinical picture showed an improvement of most parameters tested. The comparison between pre- and post-operative neurophysiological picture revealed worsening of most tested parameters. The comparison between the current sample and the Italian normative data for the SF-36 showed a worsening of physical aspects of health related quality of life; conversely there was an improvement of some mental domains. CONCLUSIONS: In our sample of LS patients the most compromised SF-36 domain was Role-Physical that measures the difficulty in every day activities due to physical problems. Conversely, the clinical findings showed a significant improvement after surgery: patients reported in particular lower sciatica after surgical treatment, but the neurophysiological evaluation did not show any improvement. PMID- 15830988 TI - Myofascial pain mimicking radicular syndromes. AB - Myofascial pain is very often underscored and misunderstood in clinical practice. In many cases the localization of myofascial pain may resemble other diseases, such as radicular syndromes (e.g., low back pain from herniated disc) and even diseases of internal organs (e.g., angina pectoris, bowel diseases or gynaecological disturbances). In pain clinics one can routinely see patients with myofascial painful disorders showing a radicular topography and normal CT and MRI: as a consequence, when vertebral abnormalities are present on CT or MRI, it should be checked whether the cause of pain is radicular, myofascial, or both. On the other hand, the conventional approach to painful disorders may lead to errors and wrong diagnosis, depending on several factors: a) pain is often considered a symptom of an organic disease; b) the diagnosis is usually directed towards the structural cause of pain only; c) the functional components of the suffering patient are underscored; d) the site of pain may introduce some bias. When the latter is concerned, it is usually admitted that a neck pain may depend on muscle contraction (e.g. torticollis), while such a cause is less commonly admitted for leg, where the attention is first directed towards the sciatic nerve; myofascial origin of pain is even less considered in abdominal or pelvic painful disorders, where patients with no structural detectable diseases are often considered as neurotic and referred to the psychiatrist. The reason for this topographical dependence of diagnosis lies in the conventional attitude to focus on the most relevant and frequent organic diseases, thus introducing a bias with relevant epistemological implications. PMID- 15830990 TI - Recognition of cemetery remains in a forensic context. AB - The purpose of this paper is to provide guidelines for the recognition of human remains from modem and historic cemeteries found in a forensic context. Three avenues of evidence may be pursued to confirm the origin of cemetery remains: context, condition of the body, and associated artifacts. This article outlines types of North American cemeteries, demonstrating how land use over time has resulted in many being closed, moved, or forgotten, leaving only the context to indicate their presence. The condition of human cemetery remains varies considerably depending on cultural practices and burial environment, but many exhibit combinations of the following traits: dried or embalmed tissue; erosion of bony pressure points; cortical bone flaking; and bone damage due to autopsy or embalming. Examples of artifact types useful in recognizing cemetery remains are also provided. Two cases from British Columbia, Canada are presented to demonstrate the diagnostic features of a disturbed cemetery burial. PMID- 15830989 TI - Prevention of recurrent radicular pain after lumbar disc surgery: a prospective study. AB - BACKGROUND: Postoperative epidural fibrosis is a major causative factor of low back pain even if microsurgical techniques are adopted to reduce this phenomenon. To prevent the recurrent radicular pain caused by this problem, we utilized adipose tissue drawn from the same patient; at the end of surgical procedure, the fat was placed around the spinal root or the dural sac. This procedure was evaluated by a prospective, randomised study. METHOD: From 180 patients operated on for lumbar discectomy between January 2000 to December 2001, 74 were enclosed in the study. In 37 patients, the spinal root was covered by autologous fat (group A), in the other 37 (control group, B), this procedure was not adopted. One year after surgery, all the patients were evaluated with clinical and radiological (Magnetic Resonance Imaging) follow-up. Only two patients were lost to follow-up. FINDINGS: 71% of the patients in group A had 100% of clinical and radiological post-operative outcome score; this result was obtained only in 35% of the patients in group B. A clinical score evaluating pain syndrome (from grade 0 to 5) and a radiological score evaluating postoperative fibrosis (from grade 0 to 4) was adopted. Therefore, group A had best outcome as compared to control group. CONCLUSIONS: The authors found a positive effect in the reconstruction of epidural fat with autograft of adipose tissue to prevent postoperative scarring and failed-back syndrome related to postoperative fibrosis. PMID- 15830991 TI - Three-dimensional cranio-facial reconstruction in forensic identification: latest progress and new tendencies in the 21st century. AB - Three-dimensional (3D) cranio-facial reconstruction can be useful in the identification of an unknown body. The progress in computer science and the improvement of medical imaging technologies during recent years had significant repercussions on this domain. New facial soft tissue depth data for children and adults have been obtained using ultrasound, CT-scans and radiographies. New guidelines for facial feature properties such as nose projection, eye protrusion or mouth width, have been suggested, but also older theories and "rules of thumbs" have been critically evaluated based on digital technology. New fast, flexible and objective 3D reconstruction computer programs are in full development. The research on craniofacial reconstruction since the beginning of the 21st century is presented, highlighting computer-aided 3D facial reconstruction. Employing the newer technologies and permanently evaluating and (re)questioning the obtained results will hopefully lead to more accurate reconstructions. PMID- 15830992 TI - Testing the reliability of frontal sinuses in positive identification. AB - The use of frontal sinus radiographs in positive identification has become an increasingly applied and accepted technique among forensic anthropologists, radiologists, and pathologists. From an evidentiary standpoint, however, it is important to know whether frontal sinus radiographs are a reliable method for confirming or rejecting an identification, and standardized methods should be applied when making comparisons. The purpose of the following study is to develop an objective, standardized comparison method, and investigate the reliability of that method. Elliptic Fourier analysis (EFA) was used to assess the variation in 808 outlines of frontal sinuses by calculating likelihood ratios and posterior probabilities from EFA coefficients. Results show that using EFA coefficient comparison to estimate the probability of a correct identification is a reliable technique, and EFA comparison of frontal sinus outlines is recommended when it may be necessary to provide quantitative substantiation for a forensic identification based on these structures. PMID- 15830993 TI - The scope of anthropological contributions to human rights investigations. AB - This paper examines the participation of anthropologists in international human rights investigations between 1990 and 1999 by surveying four of the most active organizations, including the Argentine Forensic Anthropology Team, the Guatemalan Forensic Anthropology Foundation, Physicians for Human Rights and the U.N. sponsored International Criminal Tribunal for the former Yugoslavia. The education level, sex, nationality and primary role of the anthropological members of each team are quantified, as are the types of projects in which they contributed. The results show that 134 anthropologists from 22 nations investigated nearly 1300 sites in 33 countries during the study period. While involvement is not limited to those with advanced degrees and few obstacles are placed before anthropologists who wish to participate, full-time service within these organizations is rare and those interested in a career in forensic anthropology and human rights should understand the employment limitations. PMID- 15830994 TI - Analysis of gamma-hydroxybutyric acid (GHB) in spiked water and beverage samples using solid phase microextraction (SPME) on fiber derivatization/gas chromatography-mass spectrometry (GC/MS). AB - Gamma-Hydroxybutyric acid (GHB) is a CNS depressant that has been abused recreationally for its purported euphoric and relaxation effects and for the purposes of drug facilitated sexual assault due to its sedative and amnesic effects at higher doses. The dramatic increase in the abuse of GHB and association in criminal investigations over the past decade has created the need for forensic laboratories to develop analytical methods to detect GHB in a variety of matrices. The method developed in this work used solid-phase microextraction (SPME) to extract GHB from aqueous samples followed by on-fiber derivatization and analysis by gas chromatography/mass spectrometry (GC/MS). This method detected GHB in aqueous matrices with good sensitivity, high precision, excellent linearity from 0.01 mg/mL to 0.25 mg/mL, and without the need for sample manipulation that could cause interconversion between GHB and its lactone, GBL. The method was successfully applied for detection of GHB in spiked water and beverage samples. PMID- 15830995 TI - The rapid analysis of heroin drug seizures using micellar electrokinetic chromatography with short-end injection. AB - A simple and rapid method for the analysis of heroin seizures by micellar electrokinetic chromatography with short-end injection is described. Separations were performed using an uncoated fused silica capillary, 50 cm x 50 microm I.D. x 360 microm O.D. with an effective separation length of 8 cm. The system was run at 25 degrees C with an applied negative voltage of -25 kilovolts. Injection of each sample was for 2 s at -50 mbar. UV detection was employed with the wavelength set at 210 nm. The background electrolyte consisted of 85:15 (water:acetonitrile, v/v) containing final concentrations of 25 mM SDS and 15 mM sodium borate, pH 9.5. Samples and standards were prepared in 0.1% v/v acetic acid and diluted in the run buffer containing 1 mg/ml of N,N-dimethyl-5 methoxytryptamine as an internal standard. Under these conditions a text mixture containing caffeine, paracetamol, morphine, codeine, heroin, and acetylcodeine was resolved within 1.5 min. The method was used to determine the concentration of heroin in heroin seizure samples, and the results were in good agreement with those obtained by a validated gas chromatographic method. PMID- 15830996 TI - Characterization of new miniSTR loci to aid analysis of degraded DNA. AB - A number of studies have demonstrated that successful analysis of degraded DNA specimens from mass disasters or forensic evidence improves with smaller sized polymerase chain reaction (PCR) products. We have scanned the literature for new STR loci, unlinked from the CODIS markers, which can generate amplicons less than 125 bp in size and would therefore be helpful in testing degraded DNA samples. New PCR primers were designed and tested for the STR loci D1S1677, D2S441, D4S2364, D10S1248, D14S1434, and D22S1045, arranged into two miniSTR triplexes. All loci show a moderate degree of polymorphism among 474 U.S. population samples tested and were reliable and sensitive to at least 100 pg of DNA template under controlled laboratory conditions and pristine DNA samples. The utility of these new loci were confirmed in comparing the success of the miniSTR assays for typing degraded bone samples while partial profiles were observed with the majority of the samples using a commercial STR kit. PMID- 15830997 TI - Deducing drop size and impact velocity from circular bloodstains. AB - An experimental study was done to determine the diameter and velocity of blood drops falling on a surface by measuring the size of bloodstains they produced and counting the number of radial spines projecting from them. Bloodstains were formed by releasing drops of pig blood with a range of diameters (3.0-4.3 mm) and impact velocities (2.4-4.9 m/s), onto four different flat surfaces (glass, steel, plastic, paper) with varying roughness (0.03-2.9 microm). High-speed photography was used to record drop impact dynamics. Bloodstain diameters and the number of spines formed around the rim of stains increased with impact velocity and drop diameter. Increasing surface roughness reduced stain diameter and promoted merging of spines, diminishing their number. Equations are presented that explicitly relate drop diameter and impact velocity to measurements of stain diameter and number of spines. PMID- 15830998 TI - The detection and enhancement of latent fingermarks using infrared chemical imaging. AB - The use of a new technique, Fourier transform infrared (FTIR) chemical imaging, has been demonstrated for the enhancement of latent fingermarks on a number of surfaces. Images of untreated fingermarks on glass backgrounds with excellent ridge detail were acquired using infrared chemical imaging. High quality fingermarks on glass backgrounds were also developed using ethyl cyanoacrylate (super glue) fuming and subsequent infrared chemical imaging. This new method allows the collection of images from backgrounds that traditionally pose problems for current fingermark detection methods. The background may, for example, be highly colored, have a complex pattern, or possess other pattern or image characteristics that make it difficult to separate fingermark ridges using traditional optical or luminescent visualization. One background that has proven to be a challenging surface for the development of latent fingermarks is the Australian polymer banknote. To demonstrate the power and applicability of infrared chemical imaging, fingermarks fumed with ethyl cyanoacrylate were successfully imaged from Australian polymer banknotes. PMID- 15830999 TI - Forensic mitochondrial DNA analysis of 691 casework hairs. AB - A five year retrospective review of mitochondrial DNA (mtDNA) analysis on 691 casework hairs was carried out. A full or partial mtDNA profile was obtained for > 92% of hairs. With increasing age of the hair, the likelihood of obtaining a full profile decreased, although "mini-primer sets" could often be used to capture a partial profile. With increasing color and diameter of the hair, the likelihood of obtaining a profile increased. Full or partial profiles were obtained on more than 80% of 114 hairs < or = 1.0 cm. Mixtures were observed in 8.7% of hairs tested; mixtures increased with the age of the hair and were presumed to be due to exterior surface contamination that could not be sufficiently cleaned prior to extraction, since the overall level of laboratory contamination was low. The frequency of sequence heteroplasmy was 11.4%, and both hot-spot and novel sites were observed. In about one-third of these observations, another sample in the case showed either the same heteroplasmic site or a nucleotide substitution at that site. PMID- 15831000 TI - 1H NMR analysis of GHB and GBL: further findings on the interconversion and a preliminary report on the analysis of GHB in serum and urine. AB - A 1H nuclear magnetic resonance (1H NMR) method for the determination of gamma hydroxybutyric acid (GHB) and gamma-hydroxybutyrolactone (GBL) in human serum and urine using spiked samples has been developed. The method gives linear responses (correlation coefficients of 0.99 or greater) over the concentration range 0.01 mg/mL to 4.0 mg/mL in urine and 0.3 mg/mL to 2.0 mg/mL in serum. No sample pretreatment is required. Studies of the chemical interconversion of GBL and GHB showed hydrolysis of GBL to be rapid at pH 11.54, slower and less complete (30% hydrolysis) at pH 2.54 and slowest at pH 7.0, reaching 30% hydrolysis in about 40 days. No esterification of GHB was observed at any pH. PMID- 15831001 TI - Structural determination of the principal byproduct of the lithium-ammonia reduction method of methamphetamine manufacture. AB - One common method of illicit methamphetamine manufacture utilizes an alkali metal, typically lithium, and liquid ammonia to chemically reduce ephedrine or pseudoephedrine to form methamphetamine. This method is often referred to as the lithium-ammonia reduction method or the Birch reduction method. While the hydroxyl group of ephedrine is more reactive than the aromatic ring, excess alkali metal and the presence of a proton source allow the formation of a cyclohexadiene byproduct not found in samples of methamphetamine produced from other manufacturing methods. A sample enriched in this byproduct was generated and characterized using nuclear magnetic resonance (NMR) spectroscopy, gas chromatography-mass spectrometry (GC-MS), infrared (IR) spectrophotometry, and ultraviolet (UV) spectrophotometry. The chemical structure of this byproduct was determined to be 1-(1',4'-cyclohexadienyl)-2-methylaminopropane (CMP). PMID- 15831002 TI - Determination of elemental sulfur in explosives and explosive residues by gas chromatography-mass spectrometry. AB - A new method for the positive identification of elemental sulfur in explosives and explosive residues is developed. Following a carbon disulfide wash of explosives or explosive residues, a sample of the extracted material is injected onto a gas chromatography (GC) column, then analyzed via mass-selective (MS) detection. A positive identification of elemental sulfur is based on both retention time and fragmentation pattern. The GC-MS method is demonstrated to be useful in detecting and positively identifying elemental sulfur from both burned and unburned explosive mixtures. With a detection limit of 2.5 ng (2.5 x 10(-9) grams) of elemental sulfur on the column, it is shown to be 400 times more sensitive than the presumptive chemical color test that is currently the method employed for detection of small amounts of sulfur. PMID- 15831003 TI - Analysis of explosive damage in metals using orientation imaging microscopy. AB - The goal of this project was to determine whether quantitative information concerning the size and nature of an explosive blast could be determined using Orientation Imaging Microscopy (OIM) to analyze the texture of blast-affected metal. Selected 1018 steel and 2024 aluminum samples were subjected to various explosive blasts chosen to simulate a wide range of possible pressure waves. The explosives used were PBX 9404, Comp-C4, Gelmax, and Bullseye. The explosive tests were carried out at Sandia National Laboratory, and the OIM analysis was conducted at Ames Laboratory. It was discovered that while suitable patterns could be obtained from the steel samples, the oxide layer present on the surface of the aluminum samples prevented these samples from being studied. The results of the OIM studies on the steel samples indicate that damage can be tracked using OIM imaging and that Comp-C4 seems to produce patterns significantly different than the other explosives. PMID- 15831004 TI - Person identification by gait analysis and photogrammetry. AB - Surveillance images from a bank robbery were analyzed and compared with images of a suspect. Based on general bodily features, gait and anthropometric measurements, we were able to conclude that one of the perpetrators showed strong resemblance to the suspect. Both exhibited a gait characterized by hyperextension of the leg joints, and bodily measurements did not differ by more than 6 mm on average. The latter was quantified by photogrammetry: i.e., measuring by using images of the perpetrator as captured by surveillance cameras. Using the computer software Photomodeler Pro, synchronous images from different cameras were compared and concurrent body features were identified. The program could then render the perpetrator as a three dimensional, high-precision, scalable and measurable object. PMID- 15831005 TI - On the "general acceptance" of handwriting identification principles. AB - Informed by the admissibility requirements of the broad and narrow Frye tests as well as Daubert's general acceptance factor, the present study elicited the views of a homogeneous group of forensic document examiners and a heterogeneous group of handwriting scientists regarding the degree to which a set of principles relating to the nature of handwriting and its identification were generally accepted within the respondents' fields. Among forensic document examiners, the greatest agreement was found concerning the process of examination, and somewhat less agreement on other matters. Forensic document examiners and handwriting scientists appear not to agree on the acceptability of most of the propositions. PMID- 15831006 TI - Computer-based production of comparison overlays from 3D-scanned dental casts for bite mark analysis. AB - Bite mark analysis assumes the uniqueness of the dentition can be accurately recorded on skin or an object. However, biting is a dynamic procedure involving three moving systems, the maxilla, the mandible, and the victim's reaction. Moreover, bite marks can be distorted by the anatomic location of the injury or the elasticity of the skin tissue. Therefore, the same dentition can produce bite marks that exhibit variations in appearance. The complexity of this source of evidence emphasizes the need for new 3D imaging technologies in bite mark analysis. This article presents a new software package, DentalPrint (2004, University of Granada, Department of Forensic Medicine and Forensic Odontology, Granada, Spain) that generates different comparison overlays from 3D dental cast images depending on the pressure of the bite or the distortion caused by victim biter interaction. The procedure for generating comparison overlays is entirely automatic, thus avoiding observer bias. Moreover, the software presented here makes it impossible for third parties to manipulate or alter the 3D images, making DentalPrint suitable for bite mark analyses to be used in court proceedings. PMID- 15831007 TI - Does carcass enrichment alter community structure of predaceous and parasitic arthropods? A second test of the arthropod saturation hypothesis at the Anthropology Research Facility in Knoxville, Tennessee. AB - In a second test of an arthropod saturation hypothesis, we analyzed if the on campus Anthropology Research Facility (ARF) at the University of Tennessee, Knoxville, with its 20+ yr history of carcass enrichment, is comparable to non enriched sites in community structure of predatory and parasitic arthropods that prey upon the sarcosaprophagous fauna. Over a 12-day period in June 1998, using pitfall traps and sweep nets, 10,065 predaceous, parasitic, and hematophagous (blood-feeding) arthropods were collected from freshly euthanized pigs (Sus scrofa L.) placed at ARF and at three surrounding sites various distances away (S2-S4). The community structure of these organisms was comparable in most paired site tests with respect to species composition, colonization rates, and evenness of pitfall-trap abundances on a per carcass basis. Site differences were found in rarefaction tests of both sweep-net and pitfall-trap taxa and in tests of taxonomic evenness and ranked abundances of sweep-net samples. Despite these differences, no evidence was found that the predatory/parasitic fauna at ARF was impoverished with fewer but larger populations as a result of carcass enrichment. Comparison of the sarcosaprophagous and predatory/parasitic faunas revealed a tighter (and more predictable) linkage between carrion feeders (sarcosaprovores) and their carrion than between carrion feeders and their natural enemies (predators and parasitoids), leading us to conclude that ARF is more representative of surrounding sites with respect to the sarcosaprovore component than to the predatory/parasitic component within the larger carrion-arthropod community. PMID- 15831008 TI - Hyperostosis frontalis interna: forensic issues. AB - The use of pathological conditions in age and sex determination, important factors in personal identification, is not widespread in anthropology and legal medicine. Hyperostosis frontalis interna (HFI) is a bone condition that mainly affects the inner table of the frontal bone. Although there are numerous publications on the subject, at the present time its etiology remains obscure. Several associations of symptoms, whose incidence varies according to the population studied, have been described. Age and gender appear to be linked with the preponderance of this condition, as does the presence of behavioral disturbances. The aim of our study, based on a series of 1532 autopsies, was to define the incidence and the associations observed with other pathological conditions. Thirteen cases of HFI were identified (0.8% of autopsies), 12 women and one man whose mean age was 59.15 years (range: 42-79 years). All had behavioral disturbances and most were under psychiatric care. This study emphasizes the value of this condition in medico-legal identification. PMID- 15831009 TI - Sudden infant death syndrome risk factors with regards to sleep position, sleep surface, and co-sleeping. AB - We present a study of 102 Sudden Infant Death Syndrome (SIDS) deaths using a retrospective review of medical examiner autopsy reports. The prevalence of sleep related risk factors with regards to sleep surface, sleep position, and co sleeping were determined in a population of infants less than 1-year-old. Of the 102 SIDS deaths, 67 (65.7%) were not in a crib, 63 (61.8%) were prone, and 48 (47.1%) were co-sleeping. However, 94 (92.2%) of these deaths had at least one risk factor present. Only 8 (7.8%) infants had slept alone, in a crib or bassinet, and on their back or side. Infants less than 4-months-old had a higher rate of co-sleeping (54.7%) than the older infants (25.9%), and a higher frequency of heart malformations at post-mortem examination. The older infants were more likely to exhibit pulmonary and tracheal inflammation, and neuropathology. PMID- 15831010 TI - Black soldier fly (Diptera: Stratiomyidae) colonization of pig carrion in south Georgia. AB - The black soldier fly, Hermetia illucens (L.), is thought to colonize corpses 20 30 days postmortem. However, recent observations indicate this might not be true for all cases. Therefore, we conducted a study examining colonization by the black soldier fly and other Diptera on pig carrion in a plowed field in southern Georgia from 20 September through 21 February. Our data indicate black soldier flies could colonize a corpse within the first week after death. Knowing this information could prevent a serious mistake in estimating the time at which a corpse is colonized by this species. This study also represents the first record of Chrysomya rufifacies in Georgia. PMID- 15831012 TI - Neck injuries caused by automatic two-point seat belts: an analysis of four cases. AB - Although seat belts significantly reduce the extent and severity of injuries sustained by motor vehicle occupants, seat belts are known to be associated with chest and abdominal trauma. Less commonly understood are severe neck injuries caused by the use of two-point automatic shoulder harnesses without concurrent use of a manual lap belt. Such injuries may include cervical spine fractures, craniocervical dislocations and rarely decapitation. Recognizing patterned injuries caused by seat belts and the ability to correlate autopsy findings with the circumstances surrounding the death will allow for correct interpretation of seat-belt related trauma. The four cases described detail fatal neck injuries as a result of improper seat belt use in which an automatic two-point shoulder harness was used without a manual lap restraint. In two of the cases, the victims were decapitated. PMID- 15831011 TI - Forensic dermatopathology and internal disease. AB - The gross and microscopic analysis of skin lesions at autopsy can help the pathologist understand diseases and injuries inflicted premortem, perimortem, or postmortem. From January 2003 to January 2004, skin findings at autopsy were closely examined by a dermatologist and sampled for microscopic analysis at the Southwestern Institute of Forensic Sciences. Dermatologic abnormalities in some of these cases led to the discovery of internal disease and allowed for a more complete understanding of the pathologic disease processes affecting the individual. We present four autopsy cases with skin manifestations of internal disease, including pseudoxanthoma elasticum, calciphylaxis, the sign of Leser Trelat, and papular mucinosis, and demonstrate the usefulness of the dermatological assessment at autopsy. In all cases, discovery of these skin lesions and internal disease manifestations allowed contributing factors to the death of the individual to be uncovered. PMID- 15831013 TI - Autopsy artifact created by the Revivant AutoPulse resuscitation device. AB - In certain cases, the evaluation and correct identification of resuscitative artifacts is critical to the correct diagnosis and determination of the cause and manner of death. Resuscitative artifacts can resemble homicidal or accidental injury and thus possibly be misinterpreted. Occasionally, new technologies and/or medical procedures will create original and/or distinctive artifacts. In 2003, the San Francisco Fire Department emergency personnel began field-testing the Revivant AutoPulse, an automated chest compression device. This device is currently being used in two other counties in the San Francisco Bay Area as well as regions of Florida, Virginia, and Ohio. We present three cases of resuscitative artifact that could be potentially confused with homicidal or accidental injury. These cases illustrate resuscitative artifacts, specifically lateral chest and horizontally oriented upper abdomen cutaneous abrasions created by this automated chest compression device. PMID- 15831014 TI - Fatal case of BOTOX-related anaphylaxis? AB - Anaphylactic drug reactions are rare and often serious events. The Botulinum toxin A, marketed as BOTOX, was recently approved by the Food and Drug Administration for cervical dystonia and glabellar wrinkles, after its approved use and success with blepharospasm, strabismus, and disorders of the 7th cranial nerve. It has been well received due to its efficacy in improving facial lines. This case report documents the first death associated with a Botox-lidocaine mixture given to a woman for chronic neck and back pain. Based on the medical records, autopsy, and laboratory findings, the cause of death was determined to be anaphylaxis to the Botox-lidocaine mixture. The history, indications, off label uses and possible future applications of Botox are reviewed as well as the uses and complications of lidocaine. Although the anaphylaxis cannot be definitively proven to be due to Botox alone, this case warns of an adverse reaction related to Botox, a drug that is rapidly expanding in range of use as well as increased usage. PMID- 15831015 TI - A fatal chronic ketamine poisoning. AB - A few papers in the literature reported incident deaths by acute ketamine poisoning. In this paper, we report an unusual homicide caused by chronic ketamine poisoning. The victim was a 34-year old married woman with no previous medical history (except as reported herein) who died in her own home. The court investigation revealed that she was chronically poisoned by her husband over a period of about one year in an act of homicide. Determination of ketamine concentrations in autopsy specimens was carried out with gas-chromatography/mass spectrometry (GC-MS). The results showed that ketamine concentration was 21 microg/mL in gastric contents, 3.8 microg/mL in blood and 1.2 microg/mL in urine. The most striking forensic findings were cardiac muscle fibrosis and hyaline degeneration of small arteries in victim's heart, the pathological features of ketamine poisoning previous reported only in animal studies. PMID- 15831016 TI - Investigation of class characteristics in English handwriting of the three main racial groups: Chinese, Malay and Indian in Singapore. AB - This paper investigated the class characteristics in English handwriting of the Chinese, Malays and Indians in Singapore, many of whom learned their native language as a second language. One hundred and fifty-four handwriting exemplars were collected and features such as letter designs, pen-lifts, letter spacing and embellishments were studied. A number of characteristic features peculiar to the individual racial group were identified, which confirmed the impact of their native language writing systems on English handwriting. PMID- 15831017 TI - Individuality of handwritten Arabic numerals in local population. AB - This paper reports the statistical study of writing habits for Arabic Numerals of 187 subjects in Hong Kong. A classification system of writing habits for Arabic numerals based on assigned codes of characteristic features such as slant, writing direction, relative position of strokes, angularity of turnings, shape of initial and ending strokes, etc. was developed. A set of characteristic codes representing the profile of writing habits pertaining to Arabic numerals was assigned to each writer. Apart from the distribution of characteristic features, statistical analysis of the assigned codes demonstrated homogeneity of individual hand-writing patterns. It has been shown that irrespective of the structural simplicity of Arabic numerals, no two individuals exhibited the same set of characteristic codes. The findings support the hypothesis of individuality in handwriting. PMID- 15831018 TI - One hundred seventy two deaths involving the use of oxycodone in Palm Beach County. AB - Oxycodone is a potent semi-synthetic narcotic prescribed for the management of pain. Previous investigators have reported that the abuse of oxycodone is most frequently seen in conjunction with the abuse of other drugs, although fatalities have been reported with oxycodone alone. We undertook a retrospective review of cases investigated by the Palm Beach County Medical Examiner's Office in which postmortem toxicologic studies indicated the presence of oxycodone. A total of 172 consecutive cases were studied, including 18 in which death was attributed to oxycodone toxicity, 117 to combined drug toxicity, 23 to trauma, 9 to natural causes and 5 to another drug or drugs. The postmortem blood concentrations of oxycodone overlapped among the groups. The mean blood oxycodone concentration among the cases of oxycodone toxicity was 0.69 mg/L, combined drug toxicity 0.72 mg/L and trauma 0.62 mg/L. Concentrations were lower in cases of deaths attributed to natural causes and to another drug or drugs (mean each 0.087 mg/L). Benzodiazepines, detected in 96 cases, were the most common co-intoxicants in the cases of combined drug toxicity, followed by cocaine, which was found in 41. The most frequently encountered benzodiazepine was alprazolam. This study confirms that deaths in which oxycodone is a factor are most commonly cases of combined drug toxicity. The high incidence of alprazolam as a co-intoxicant has not been previously recognized. PMID- 15831019 TI - Pancuronium bromide (Pavulon) isolation and identification in aged autopsy tissues and fluids. AB - The isolation and detection of pancuronium bromide was developed for aged autopsy samples to identify and confirm this compound in questioned tissue samples. A novel protocol was optimized for the isolation of the target drug in highly decomposed tissues. Solid-phase extraction (SPE) cartridges containing styrene divinylbenzene were investigated. This polymer retained quaternary drugs and facilitated sequential elution upon washing with commonly available solvents. The semi-purified SPE samples were prescreened by pyrolysis GC-MS. A candidate specimen was then confirmed by microbore high-performance liquid chromatography/electrospray-ionization/mass spectrometry (microHPLC-ESI-MS/MS) with a triple-quadrupole mass spectrometer. The developed procedures provided a qualitative or semiquantitative (at best) basis for the investigation of difficult cases involving overdoses of polar drugs. PMID- 15831020 TI - Postmortem production of ethanol in different tissues under controlled experimental conditions. AB - The aim of this study was to follow the postmortem ethanol production phenomenon under controlled experimental conditions (temperature, time interval) in different tissues. Specimens of blood, liver, skeletal muscle and kidney were taken from 30 corpses and no chemical preservatives were used in the specimens collected. Ethanol concentrations were detected by gas chromatography. All specimens stored at -20 degrees C and 4 degrees C did not show any change in ethanol concentration in an eight-day time interval. At 20 degrees C and 30 degrees C, all tissues, except blood, showed statistically significant ethanol production over the time interval tested. However, blood sample kept at 30 degrees C, showed statistically significant increase in ethanol production on the 2nd and 4th day comparing to the controls. Thus, we can state that postmortem ethanol production occurs in different tissues, and is increased at higher temperatures and, in general, it is in accordance with the course of time. PMID- 15831021 TI - Disposition of quetiapine in biological specimens from postmortem cases. AB - Quetiapine is a new atypical antipsychotic that was approved in 1997 by the U.S. Food and Drug Administration for the treatment of schizophrenia. It possesses a high affinity for 5-HT2 receptors and a low affinity for D1 and D2 dopamine receptors. Because quetiapine has only been released recently to the U.S. market, little information exists regarding therapeutic, toxic, and lethal concentrations. This study reports the detection of quetiapine in 13 postmortem cases. Following a basic liquid-liquid extraction, quetiapine was identified and quantitated by capillary gas chromatography with nitrogen phosphorus detection. Confirmation was accomplished by full scan electron impact gas chromatography/mass spectrometry. Heart blood quetiapine concentrations ranged from 0.07 to 18.37 mg/L (N = 12, mean +/- SD = 3.42 +/- 5.67, median 0.62) and femoral blood concentrations ranged from 0.06 to 19.25 mg/L (N = 10. mean +/- SD = 3.89 +/- 6.12, median 0.81). The average heart blood/femoral blood ratio was 1.31 (range 0.55 to 2.57, N = 10). Urine, bile, and gastric contents were assayed in all cases in which they were submitted. In three cases, the cause of death was determined to be quetiapine toxicity. In these cases heart blood concentrations ranged from 0.72 to 18.37 mg/L (N = 3). These data may provide a basis for establishing levels associated with quetiapine toxicity as well as therapeutic concentrations in postmortem specimens. PMID- 15831023 TI - The polymorphisms of four Y-chromosome short tandem repeat loci in Chinese and Japanese populations. PMID- 15831022 TI - The application of pancuronium bromide (Pavulon) forensic analyses to tissue samples from an "Angel of Death" investigation. AB - The case report of a serial killer who worked at several hospitals as a respiratory therapist is presented. The suspect was initially labeled a benevolent Angel of Death who ended the suffering of elderly patients through mercy killing. However, his subsequently declared motive for homicide was very different from other similar cases in medical settings. The application of new analysis techniques for the detection of pancuronium bromide in a series of aged exhumation tissues gave positive results and led to the resultant conviction of the therapist. PMID- 15831024 TI - Population data for the twelve Y-chromosome short tandem repeat loci from the sample of multinational population in Bosnia and Herzegovina. PMID- 15831025 TI - Allele frequency distribution of STR loci D5S814 in four populations. PMID- 15831026 TI - Allele frequency distribution of STR loci D5S1486 in three populations. PMID- 15831027 TI - Allele frequency distributions for 15 STR loci in Chinese Chengdu Han population. PMID- 15831028 TI - Polymorphism data at D8S315 locus among five ethnic groups of Kerala from South India. PMID- 15831029 TI - Polymorphism data at AY639919 and AY639922 loci in Chinese population. PMID- 15831030 TI - Polymorphism of three STR loci in Chinese population. PMID- 15831031 TI - Mexican population data on fifteen STR loci (Identifiler kit) in a Chihuahua (North Central Mexico) sample. PMID- 15831032 TI - Y-chromosomal haplotypes for the PowerPlex Y for twelve STRs in a Peruvian population sample. PMID- 15831033 TI - Allele frequencies for fifteen STR loci in Guinea-Bissau population. PMID- 15831034 TI - Genetic profile of five STR loci D2S1338, D8S1179, D14S306, D19S253 and D18S535 in a Chinese population. PMID- 15831035 TI - Proposed pharmaceutical regulations for skeletal muscle depolarization drugs. PMID- 15831036 TI - [The perioperative treatment of hypopharyngeal carcinoma with cervical esophageal invasion]. AB - OBJECTIVE: To present our experiences of perioperative therapy on 7 patients suffered from hypopharyngeal carcinoma with cervical esophageal invasion. METHOD: Among the 7 patients, 6 patients had unilateral functional neck dissection, total laryngoesophagectomy and reconstruction with gastric pulling-up. One patient had gastrectomy, reconstruction with colonic pulling-up as well as radical neck dissection, total laryngoesophagectomy. Five of 7 patients have unilateral partial thyroidectomy. RESULT: All patients complained of vomiting and other abnomalities of digestion, 1 case complicated with pharyngocutaneous fistula, 1 patient developed acute renal failure, postoperatively. CONCLUSION: It is very important to improve theraputic skill during perioperative period,which can reduce complications of advanced hypopharyngeal and cervical esophageal cancer and increase patients' survival. PMID- 15831037 TI - [Patterns and prognostic factors of cervical lymph node metastasis in supraglottic laryngeal carcinoma]. AB - OBJECTIVE: To investigate the risk factors in the prediction of cervical lymph node metastases and the cervical lymph node prognostic factors in supraglottic laryngeal carcinoma. METHOD: A retrospective study was carried out to review the histopathological data from 55 supraglottic laryngeal carcinoma patients. The relationship between histopathological parameters and cervical lymph node metastases were evaluated by means of a univariate chi2 test and multivariate stepwise logistic regression model. And the Cox regression model was used to define possible pathological parameters of cervical lymph node affecting survival including N staging, presence of cervical lymph node metastases and extracapsular nodal spread, size and number of positive neck nodes, and levels of positive neck nodes. RESULT: In a univariate analysis, it was confirmed that the following variables correlated to cervical lymph node metastases, i.e., depth of primary tumor infiltration, size and histological grade of primary tumor. In a multivariate analysis, the most significant risk factors for cervical lymph node metastases were the size and histological grade of primary tumor. The overall 5 year survival rate of patients with supraglottic laryngeal carcinoma was 52.7%. In a Cox regression analysis, N staging, size of positive neck nodes, presence of cervical lymph node metastasis and extracapsular nodal spread were found to be related to the prognosis of supraglottic laryngeal carcinoma. CONCLUSION: Cervical lymph node metastasis is one of the most significant prognostic factors of supraglottic laryngeal carcinoma. The identification of patients at risk for cervical lymph node metastasis and the management of the neck by coping with pathological factors of neck node affecting survival are crucial steps to improve the effect of supraglottic laryngeal carcinoma treatment. PMID- 15831038 TI - [Suppression subtractive hybridization to identify differentially expressed gene between laryngeal tumor and the normal tissue adjacant to the tumor]. AB - OBJECTIVE: To screen the differently expressed genes between laryngeal tumor and the normal tissue adjacant to the tumor and construct the subtractive cDNA library. METHOD: The subtractive cDNA library was contructed with the suppression subtractive hybridization (SSH) technique. Nine random clones are made as probes for the reverse Northern hybridization, after sequenced. The sequences are compared with known sequences using blastn against GenBank across the Internet. RESULT: Except one clone ligated by two adaptor, the others represented six gene and one IgG heavy chain constant region. Among the six genes, one was a new kind of antibody whose function was not understood,the others stood for genes as follows: keratin, beta-actin, myosin, tumor necrosis factor, alkaline phosphatase. CONCLUSION: Screening the differently expressed gene with SSH is reliable. These genes play a pivotal role in tumorigenesis, tumor promotion, tumor metastasis, tumor immunity and tumor marker. PMID- 15831039 TI - [Treatment for obstructive sleep apnea-hypopnea syndrome with coblator plasma surgery]. AB - OBJECTIVE: To explore the method of coblator plasma surgery and modified uvulopalatopharyngoplasty with uvula reserved and tongue base operation and its effect on obstructive sleep apnea-hypopnea syndrome (OSAHS). METHOD: All 56 cases were operated on infraturbinal, tongue base, uvula, soft palate and/or tonsil with coblator plasma surgery, the uvula was completely reserved by modified uvulopalatopharyngoplasty in 9 cases. RESULT: The 6-months responders were 87.5% respectively. Five cases suffered from pharyngeal paraesthesia, 2 cases suffered from stomatolalia, 1 case suffered from pharyngeal secondary hemorrhage. CONCLUSION: The coblator plasma surgery on OSAHS is an effective and safe method. PMID- 15831040 TI - [Expression of MDM2 and p27 in hypopharyngeal carcinoma and their clinical significance]. AB - OBJECTIVE: To explore the relationship between the expression of MDM2 and p27 in hypopharyngeal carcinoma and clinicopathological parameters and their clinical significance. METHOD: The expression of MDM2 and p27 in 59 hypopharyngeal carcinoma were detected by S-P immunohistochemical technique. RESULT: The high expression rates of MDM2 and p27 were 66.1% and 33.9%, respectively, and the low expression rates of them were 33.9% and 66.1%, respectively. The expression of MDM2 was unrelated to p27 (P > 0.05). The lymph node metastasis in high expression group of MDM2 was significantly higher than that in low-expression group (P < 0.01). The expression rate of p27 was significantly different among the four T groups and degree of differentiation, respectively (P < 0.05). The survival rate was significantly higher in high-expression rate group of MDM2 than that in low-expression rate group (P < 0.01), while the survival rate was significantly lower in high-expression rate group of p27 than that in low expression rate group (P < 0.05). CONCLUSION: The expression of MDM2 and p27 may be involved in carcinogenesis and progression in hypopharyngeal carcinoma. Combined detecting of MDM2 and p27 may predict its prognosis better in hypopharyngeal carcinoma. PMID- 15831041 TI - [Applied anatomic study for genioglossus advancement]. AB - OBJECTIVE: To provide anatomic basis for genioglossus advancement. METHOD: Forty adult cadaver heads (male=29, female=11) were dissected to measure genial tubercles and observe the start-stop points of genioglossus muscle and geniohyoid muscle. RESULT: The genioglossus muscle originates from the superior mental spine, and the geniohyoid muscle originates from the inferior mental spine. The height, width and process of the superior mental spin were (5.82 +/- 0.71)mm, (6.98 +/- 1.35)mm, (2.92 +/- 0.97)mm respectively, while (5.97 +/- 1.39)mm, (3.81 +/- 0.52)mm, (0.77 +/- 0.62)mm in the inferior mental spine. The distance from the menton to the inferior edge of the inferior mental spine was (5.11 +/- 1.33)mm. The thickness of the mandible at the genial tubercle was (11.95 +/- 1.59)mm in total. The distance from the apices of the incisors to the superior edge of the superior mental spine was (15.57 +/- 1.82)mm in male, (9.36 +/- 2.79)mm in female, which had statistically difference (t = 6.21, P < 0.01). There are statistical difference at width, process between superior mental spine and inferior mental spine,not at height. CONCLUSION: The distance from the apices of the incisors to the superior edge of the superior mental spine in male is greater than that in female. When the genioglossus and geniohyoid are advanced simultaneously, the inferior horizontal bone cut should be approximately 5 mm above the menton, which should be 11 mm if the genioglossus is advanced single. The superior horizontal bone cut should be approximately 18mm above the menton. The lateral vertical cut should be 4 mm lateral to midline. It is visible, practicable and exact that the osteotomy is located with the menton and midline as marks. PMID- 15831042 TI - [The expression of VEGF and iNOS in laryngeal squamous cell carcinoma and its clinical significance]. AB - OBJECTIVE: To investigate the mRNA expression of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) in laryngeal squamous carcinoma and its correlation with clinicopathologic parameters, to identify the interactions between the expression of VEGF and iNOS. METHOD: The mRNA expression of VEGF and iNOS in fresh laryngeal squamous carcinoma tissues and adjacent normal tissues from 40 patients were examined by reverse transcription-polymerase chain reaction. RESULT: The positive expression rate of VEGF and iNOS in tumor samples were significantly higher than that in normal tissues (P < 0.01); in tumor samples, their expression were not associated with age, sex, tumor site and T stage (P > 0.05); their expression were higher in moderately and poorly differentiated tumors than in well differentiated tumors (P < 0.05); their expression were significantly higher in tumors with cervical lymph node metastasis than in tumor without cervical lymph node metastasis (P < 0.01). There were correlation between the expression of VEGF and iNOS in tumor samples (P < 0.05). CONCLUSION: VEGF and iNOS could up-regulate each other to promote the development and cervical lymph node metastasis of laryngeal squamous carcinoma. PMID- 15831043 TI - [The maxillary swing approach to resect the tumors in the skull base and the paranasopharyngeal space]. AB - OBJECTIVE: To determine the best approach to resecting the tumors in paranasopharyngeal space and skull base. METHOD: Through a Weber-Ferguson Longmire incision,the whole maxilla was freed from bony connections and swung laterally while remaining attached to the masseter muscle and the cheek flap. Then the skull base and paranasopharyngeal space was exposed. So the tumor could be resected completely. Two cases of tumor in paranasopharyngeal space, 1 case of tumor in skull base, by the maxillary swing approach, the tumor was resected under sight. The fat of check was used to pack the space. RESULT: One case was vascular smooth muscle tumor and 1 case was adenoid cystic carcinoma in paranasopharyngeal space, 1 case was chordoma in skull base. The last case received post-operation radiotherapy. All the cases healed with no severe dentofacial malformation and dysfunction. In the follow up period, no one recurred postoperatively. CONCLUSION: The maxillary swing approach to resect the tumor in the skull base and the paranasopharyngeal can expose better. The operation is safe with little damage. PMID- 15831044 TI - [Expression and significance of PTEN and caspase-3 in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To elucidate the expression and significance of PTEN and Caspase-3 in laryngeal squamous cell carcinoma. METHOD: Immunochemical method was used to study 289 cases of laryngeal carcinoma, including 60 cases atypical hyperplasia of vocal fold and 20 cases of normal laryngeal tissues. RESULT: The PTEN gene was expressed in 58.8% laryngeal carcinoma tissues, in 86.7% atypical hyperplasia of vocal fold and 100.0% normal laryngeal tissues. The differences were significance (P < 0.05). The Caspase-3 was expressed in 48.1% laryngeal carcinoma tissues, in 61.7% atypical hyperplasia of vocal fold and in 85.0% normal laryngeal tissues. The differences were significance (P < 0.05). In addition, there was a positive correlation between the expression of Caspase-3 and PTEN (P < 0.01). CONCLUSION: Expression of PTEN and Caspase-3 in laryngeal carcinoma is different from that of normal laryngeal tissues, which may play an important role in the tumorigenesis and development of laryngeal carcinoma. PMID- 15831045 TI - [Characteristics of respiratory disturbance during REM sleep in OSAHS]. AB - OBJECTIVE: To study the characteristics of respiratory disturbance during REM sleep in OSAHS through the signals surveyed by polysomnography (PSG). METHOD: The result of sleeping monitoring of 180 cases with OSAHS by PSG was reviewed, the AHI and the RIT index was used to evaluate the severity in different sleep stage as in REM and N-REM stage to find out the characteristics of the respiratory disturbance. RESULT: The study showed that AHI index in mild group increased greatly in REM period (P < 0.05), but no significant changes between the moderate and severe groups. And the lowest oxygen saturation (LSaO2) was lower during REM stage than that during N-REM stage (P < 0.05). CONCLUSION: (1) The time of REM in OSAHS patients often decreases according to their severity. (2) The respiratory disturbance during the stage REM is much severe than during the stage N-REM. The main manifestation is that the time of apnea-hypopnea is more longer and the AHI index does not significantly change. (3) The RIT index is more closely related to the severity of respiratory disturbance than AHI index. PMID- 15831046 TI - [The study of the expression, effect and clinical relationship of TGF-beta1 CD44V6 bcl-2 and p53 in the larynx squamous carcinoma]. AB - OBJECTIVE: This study was to measure the expression of TGF-beta1 in larynx squamous carcinoma, to investigate the relationship between TGF-beta1 and the expressions of CD44V5 , bcl-2 and p53, and the relationship between TGF-beta1 and pathological grading of tumours, the clinical staging and the survival rate of the patients, through which the effect of TGF-beta1 on the pathogenesis of the larynx squamous carcinoma was to be known. METHOD: With immunohistochemistry, the expressions of TGF-beta1, CD44V6, bcl-2 and p53 were measured in the paraffin sections of the larynx squamous carcinoma tissues of 43 patients. RESULT: In pathological grading I, II, III, respectively, the positive expression ratios of TGF-beta1 were 86.96%, 82.35%, 100.00%; CD44V6 were 60.87%, 70.59%, 100.00%; bcl 2 were 82.61%, 94.12%, 100.00%; p53 were 26.09%, 35.29%, 66.67%. CONCLUSION: (1) TGF-beta1 had some relationships to the pathological grading, the clinical staging, the metastasis to cervical lymph node, the survival rates of patients in 3 and 5 years. TGF-beta1 can be used as a marker to evaluate the development and predict the metastasis of tumors and the prognosis of patients. (2) The expression of TGF-beta1 in larynx carcinoma had some correlation to CD44V6, bcl-2 and p53. PMID- 15831047 TI - [The effects of body posture on upper airway in OSAHS patients]. AB - OBJECTIVE: To explore the effects of body posture on upper airway shapes in OSAHS patients. METHOD: Thirty-one male OSAHS patients were included in the study. The fibre endoscopic photos were taken on palate and lingua region upper airway at normal breath at supine with 30 degree neck flexion, supine and right lateral recumbent position, respectively. The cross section area and the dimensions of palate and lingua region upper airway were studied by running image tools software in computer. RESULT: In OSAHS patients, there were obviously difference in the area and arrowy dimension of palate airway among three posture, and so to in area, arrowy and coronary dimension of lingua airway. CONCLUSION: The study suggests that the body posture changes have obviously influence on upper airway, and the influence is the base of apnea increasing at special posture. PMID- 15831048 TI - [EGG and acoustic test for the diagnosis of laryngeal diseases]. AB - OBJECTIVE: To explore the relationship between electroglottography/acoustic test parameter and disease diagnosis. METHOD: 3.0 software by Dr. Speech was used to test synchronically the electroglottography and acoustic test parameter of 503 cases so as to make a statistical analysis and comparison. RESULT: The shimmer was similar with the jitter on the inspection of polyp of vocal cord and vocal nodules, but the shimmer was more susceptive than the jitter. Compared with the healthy group, mean F0, jitter and shimmer of polyp of vocal cord increased, while the Harmonic-to-Noise decreased, so did the vocal nodules and carcinoma of larynx. Compared with the normal, the contact index of carcinoma of larynx increased, but the contact quotient remained on the same level. CONCLUSION: EGG, acoustic test number comparison and wave change provide an objective quantized index for the diagnosis of laryngeal diseases. PMID- 15831049 TI - [Multivariate analysis of the parameters related to recurrence of laryngeal papilloma in adult]. AB - OBJECTIVE: To investigate the clinical and pathological characteristics of astrocytoma and the factors related to pathological gradings as well as recurrence in laryngeal papilloma in adult. METHOD: Fifty samples from patients were selected and 10 cases of normal tissue or inflammation tissue were used as control group. The expression of Pgp, GST-pi, Topo II and Ki-67 and VEGF by immunohistochemical stain and the microvessel density by morphometry were studied. RESULT: (1) The positive rates of Pgp, GST-pi and Topo II were not correlated with the age, sex, tumor size and the recurrence (P > 0.05). (2) Both recurrent tissue and current tissue showed various positivity for Ki-67. The difference were significant (P < 0.01). Taking 39% of Ki-67 expression as threshold, there was significant difference between two curves indicating favorable recurrence in the tumor group respectively (P < 0.01). (3) Microvascular changes,such as the density of blood vessels demonstrated significantly difference between the two tumor group (P < 0.01). An average vessel count of more than 19 (x 200) suggested an easier recurrence. CONCLUSION: Use of the cox-regression model in multivariate analysis shows that there are five parameters closely related to the recurrence of laryngeal papilloma in adult, of which the Ki-67 is the most critical factor, second the microvessel density, while Pgp, Topo II and GST-pi in tumor are independent favorable factors for laryngeal papilloma of adult. PMID- 15831050 TI - [Expression and significance of nuclear factor kappaB in laryngeal carcinoma]. AB - OBJECTIVE: To explore the expression and significance of NF-kappaB in laryngeal carcinoma. METHOD: The protein expression of NF-kappaB p65 in laryngeal carcinoma tissue and normal laryngeal tissue was examined by Western blot; NF-kappaB DNA binding activity was examined by electrophoretic motility shift assay (EMSA) in 20 laryngeal carcinoma tissue and 12 normal laryngeal tissue. RESULT: The levels of NF-kappaB p65 protein and NF-kappaB DNA binding activity in the laryngeal carcinoma tissue were higher than those in normal laryngeal tissue. CONCLUSION: NF-kappaB may be an important oncoprotein and involves in the occurrence and development of laryngeal carcinoma. PMID- 15831051 TI - [Effect of indomethacin on the growth and invasion of Hep-2 cells in human laryngeal cancer]. AB - OBJECTIVE: To assess the effect of indomethacin on the growth and invasion of Hep 2 cell line in human laryngeal cancer in vitro. METHOD: Hep-2 cell line was exposed to indomethacin at different concentration for 24 and 48 h. Then the cells were counted by trypan blue exclusion test for determining the viability of cells. And the cell DNA analysis was conducted with flow cytometry. Cell morphology was observed by phase contrast microscope and transmission electron microscope. Hep-2 cells were exposed to indomethacin at different concentration for 48 h. Then the cell growth rate, the colonies formation in soft agar medium and cell mobility were examined. Monolayer invasion assay gave us cell invasion index. RESULT: The alive cells after indomethacin treatment were reduced to a dose and time-dependent situation. And the cells appeared a significant peak of apoptosis, in flow cytometry analysis. Hep-2 cells were smaller and longer than in control; dark cells also can be seen. Dealed with indomethacin, the colonies formed of Hep-2 cells were fewer than that of control; the cell mobility were weakened and the invasion index were decreased,but cells growth rate weren't changed. CONCLUSION: Indomethacin could inhibit the growth and invasion of Hep-2 cells in vitro. PMID- 15831052 TI - [Objective multi-parameters analysis of the voice dysphonia]. AB - OBJECTIVE: The purposes of this paper are to develop a multi-parametric protocol, which can evaluate the voice objectively and quantitatively,and to investigate its correlation with the perceptual analysis. METHOD: The voice samples were collected from 83 patients with dysphonia and 40 subjects with normal voices. All the subjects were women. The objective parameters, including fundamental frequency (F0), jitter, Shimmer, fundamental frequency standard deviation (F0SD), fundamental frequency tremor, amplitude tremor, normalized noise energy (NNE), harmonic-to-noise ratio(HNR), signal-to-noise ratio(SNR) and maximal phonatory time (MPT), which were measured mainly on a sustained vowel /a/, were recorded on a software named Dr. Speech for Windows. According to the G component of the GRBAS, the voice was graded from 0 for normal to 3 for severe dysphonia. RESULT: Using the discriminate analysis, a five parameters protocol (MPT, jitter, NNE, HNR and shimmer) was developed and it showed that this protocol allowed 79.8% concordance with jury classification. CONCLUSION: The voice objective analysis should be multi-parameters. Our multi-parameters evaluation protocol is able to reflect the result of perceptual analysis. PMID- 15831054 TI - Dengue haemorrhagic fever, Timor-Leste--update. PMID- 15831053 TI - [Skull base reconstruction and rehabilitation]. AB - OBJECTIVE: To review our experience with skull base reconstruction and rehabilitation. METHOD: The clinical data of 30 patients with skull base reconstruction from 200 patients performed skull base surgery were retrospectively reviewed. Bone defect of skull base were repaired in 20 patients and dura defect in 6 patients and soft tissue defect in infratemporal fossa in 8 patients. RESULT: Postoperatively there were transient cerebrospinal leakage in 3 patients, and non-symptom intracranial pneumatosis in 3 patients, and epidural hematoma in 1 patient, and meningitis in 1 patients. CONCLUSION: Immediate reconstruction of skull base with suitable material and surgical technique can prevente complications such as postoperative infection and cerebrospinal leakage brain hernation. PMID- 15831055 TI - Update on Lassa fever in West Africa. PMID- 15831056 TI - Retrospective rubella review, Maldives. PMID- 15831058 TI - Cyclo-oxygenase-2 inhibitors : beneficial or detrimental for athletes with acute musculoskeletal injuries? AB - The major goal of clinicians when treating acute musculoskeletal injuries is to return athletes to their pre-injury level of function, ideally in the shortest time possible and without compromising tissue-level healing. In this regard, a commonly used intervention is the taking of NSAIDs. These are used to limit the amount and duration of inflammation, and to control pain. While NSAIDs have become synonymous with the management of acute musculoskeletal injuries, their efficacy has yet to be proven. This is of particular concern in view of recent research investigating the latest class of NSAIDs - selective cyclo-oxygenase-2 inhibitors (COXIBs). COXIBs were developed to reduce the adverse gastrointestinal (GI) effects of traditional NSAIDs. While they have beneficial anti-inflammatory and analgesic properties, and appear to facilitate earlier return to function following acute injury, the effect of COXIBs on tissue-level healing is currently unknown. In experimental animal models of acute injury, COXIBs have been shown to be detrimental to tissue-level repair. Specifically, they have been shown to impair mechanical strength return following acute injury to bone, ligament and tendon. Clinically, this may have implications for ongoing morbidity and future injury susceptibility. However, the current animal studies have limited translation to the clinical setting, particularly because of significant limitations relating to drug use and dosage in these studies. There is currently no randomised, controlled trial evidence of the tissue-level effects of COXIBs on acute musculoskeletal injuries. In addition to questions relating to the effect of COXIBs on tissue-level healing, further questions regarding the use of these agents have been raised given a recent link being shown between one COXIB (rofecoxib) and an increased risk for adverse cardiovascular events. Whether this finding is related to the individual properties of rofecoxib or is a class phenomenon is the subject of ongoing investigation. However, in light of the potential risks associated with using COXIBs, an acceptable and possibly safer alternative in the management of acute musculoskeletal injuries may be to use traditional NSAIDs. Traditional NSAIDs do carry the potential for greater adverse GI effects and their clinical effects on tissue-level healing remain relatively unknown. However, they do not appear to be associated with adverse cardiovascular effects, and they are effective pain relievers and cheaper alternatives. PMID- 15831059 TI - The science of cycling: physiology and training - part 1. AB - The aim of this review is to provide greater insight and understanding regarding the scientific nature of cycling. Research findings are presented in a practical manner for their direct application to cycling. The two parts of this review provide information that is useful to athletes, coaches and exercise scientists in the prescription of training regimens, adoption of exercise protocols and creation of research designs. Here for the first time, we present rationale to dispute prevailing myths linked to erroneous concepts and terminology surrounding the sport of cycling. In some studies, a review of the cycling literature revealed incomplete characterisation of athletic performance, lack of appropriate controls and small subject numbers, thereby complicating the understanding of the cycling research. Moreover, a mixture of cycling testing equipment coupled with a multitude of exercise protocols stresses the reliability and validity of the findings. Our scrutiny of the literature revealed key cycling performance determining variables and their training-induced metabolic responses. The review of training strategies provides guidelines that will assist in the design of aerobic and anaerobic training protocols. Paradoxically, while maximal oxygen uptake (V-O(2max)) is generally not considered a valid indicator of cycling performance when it is coupled with other markers of exercise performance (e.g. blood lactate, power output, metabolic thresholds and efficiency/economy), it is found to gain predictive credibility. The positive facets of lactate metabolism dispel the 'lactic acid myth'. Lactate is shown to lower hydrogen ion concentrations rather than raise them, thereby retarding acidosis. Every aspect of lactate production is shown to be advantageous to cycling performance. To minimise the effects of muscle fatigue, the efficacy of employing a combination of different high cycling cadences is evident. The subconscious fatigue avoidance mechanism 'teleoanticipation' system serves to set the tolerable upper limits of competitive effort in order to assure the athlete completion of the physical challenge. Physiological markers found to be predictive of cycling performance include: (i) power output at the lactate threshold (LT2); (ii) peak power output (W(peak)) indicating a power/weight ratio of > or =5.5 W/kg; (iii) the percentage of type I fibres in the vastus lateralis; (iv) maximal lactate steady-state, representing the highest exercise intensity at which blood lactate concentration remains stable; (v) W(peak) at LT2; and (vi) W(peak) during a maximal cycling test. Furthermore, the unique breathing pattern, characterised by a lack of tachypnoeic shift, found in professional cyclists may enhance the efficiency and metabolic cost of breathing. The training impulse is useful to characterise exercise intensity and load during training and competition. It serves to enable the cyclist or coach to evaluate the effects of training strategies and may well serve to predict the cyclist's performance. Findings indicate that peripheral adaptations in working muscles play a more important role for enhanced submaximal cycling capacity than central adaptations. Clearly, relatively brief but intense sprint training can enhance both glycolytic and oxidative enzyme activity, maximum short-term power output and V-O(2max). To that end, it is suggested to replace approximately 15% of normal training with one of the interval exercise protocols. Tapering, through reduction in duration of training sessions or the frequency of sessions per week while maintaining intensity, is extremely effective for improvement of cycling time-trial performance. Overuse and over training disabilities common to the competitive cyclist, if untreated, can lead to delayed recovery. PMID- 15831060 TI - The science of cycling: factors affecting performance - part 2. AB - This review presents information that is useful to athletes, coaches and exercise scientists in the adoption of exercise protocols, prescription of training regimens and creation of research designs. Part 2 focuses on the factors that affect cycling performance. Among those factors, aerodynamic resistance is the major resistance force the racing cyclist must overcome. This challenge can be dealt with through equipment technological modifications and body position configuration adjustments. To successfully achieve efficient transfer of power from the body to the drive train of the bicycle the major concern is bicycle configuration and cycling body position. Peak power output appears to be highly correlated with cycling success. Likewise, gear ratio and pedalling cadence directly influence cycling economy/efficiency. Knowledge of muscle recruitment throughout the crank cycle has important implications for training and body position adjustments while climbing. A review of pacing models suggests that while there appears to be some evidence in favour of one technique over another, there remains the need for further field research to validate the findings. Nevertheless, performance modelling has important implications for the establishment of performance standards and consequent recommendations for training. PMID- 15831062 TI - Inhibition of saccade and vergence eye movements in 3D space. AB - Inhibitory capacity was investigated by measuring the eye movements of normal subjects asked to fixate a central point, and to suppress eye movements toward visual distracters appearing in the periphery or in depth. Eight right-handed young adults performed such a suppression or distracter task. In different conditions, the distracter could appear at 10 degrees left or right at a distance of 20, 40, or 150 cm (calling for horizontal saccades), or in a central position far or close (calling for convergence or divergence), or 7.5 degrees up or down at 40 or 150 cm (calling for vertical saccades). Eye movements were recorded binocularly with an infrared light eye-movement device. Results showed that (1) suppression performance was not perfect, as the subjects still produced eye movements; (2) errors were distributed unequally in three-dimensional space, with more frequent errors toward distracters calling for convergence, or leftward and downward saccades at a close distance; (3) distracters calling for saccade suppression yielded saccades in the direction of the distracter (that we called prosaccades), and saccades directed away from it (that we called spontaneous antisaccades); (4) for vergence, only distracters calling for convergence yielded errors, which were always promovements; (5) in addition, a small convergent drift was found for convergence distracters. Differences in the errors between saccade and vergence suggest that different inhibitory mechanisms may be involved in the two systems. Spatial left/right, up/down, and close/far asymmetries are interpreted in terms of attentional biases. PMID- 15831063 TI - Luminance-color correlation is not used to estimate the color of the illumination. AB - Humans can identify the colors of objects fairly consistently, despite considerable variations in the spectral composition of the illumination. It has been suggested that the correlation between luminance and color within a scene helps to disentangle the influences of illumination and reflectance, because the surfaces that reflect the light of the illuminant well will normally be bright. Because the reliability of the luminance-color correlation as an indicator of the chromaticity of the illuminant depends on the number of surfaces that are considered, we expected the correlation to be determined across large parts of the scene. To examine whether this is so, we compared different scenes with matched luminance and chromaticity, but in which the correlation between luminance and chromaticity was manipulated locally. Our results confirm that there is a bias in perceived color away from the chromaticity of bright surfaces. However, the results show that only the correlation within about 1 degree of the target is relevant. Thus, it is unlikely that the visual system uses the correlation between luminance and color to explicitly determine the chromaticity of the illuminant. Instead, this correlation is presumably implicitly considered in the way that the color contrast at borders is determined. PMID- 15831064 TI - Economy of scale: a motion sensor with variable speed tuning. AB - We have previously presented a model of how neurons in the primate middle temporal (MT/V5) area can develop selectivity for image speed by using common properties of the V1 neurons that precede them in the visual motion pathway (J. A. Perrone & A. Thiele, 2002). The motion sensor developed in this model is based on two broad classes of V1 complex neurons (sustained and transient). The S-type neuron has low-pass temporal frequency tuning, p(omega), and the T-type has band pass temporal frequency tuning, m(omega). The outputs from the S and T neurons are combined in a special way (weighted intersection mechanism [WIM]) to generate a sensor tuned to a particular speed, v. Here I go on to show that if the S and T temporal frequency tuning functions have a particular form (i.e., p(omega)/(m(omega) = k/omega), then a motion sensor with variable speed tuning can be generated from just two V1 neurons. A simple scaling of the S- or T-type neuron output before it is incorporated into the WIM model produces a motion sensor that can be tuned to a wide continuous range of optimal speeds. PMID- 15831065 TI - Suppression of monocular visual direction under fused binocular stimulation: evoked potential measurements. AB - Visual evoked potentials (VEPs) were recorded in response to a vernier onset/offset target presented to one eye that was combined with matching static targets in the other eye. The monocular response was dominated by a negative peak at 160 ms that occurred after a set of offsets was introduced into a one dimensional random bar pattern. The static targets produced no discernible VEP response by themselves, but when fused binocularly with the oscillating vernier target, they produced shifts in perceived visual direction that influenced the VEP response. A vernier target fused with static vertical bars was perceived to alternate in depth between a flat surface and one broken into two interleaved surfaces. The response to this "surface-breaking" was as large or larger than the response to the monocular vernier offset. This response was much reduced when the oscillating vernier was fused with a static offset vernier (5' offset) that produced a percept of segregated regions moving in depth. Apparently, the VEP is strongly driven by shifts in visual direction that alter surface, texture, or contour contiguity. PMID- 15831061 TI - Hormonal responses and adaptations to resistance exercise and training. AB - Resistance exercise has been shown to elicit a significant acute hormonal response. It appears that this acute response is more critical to tissue growth and remodelling than chronic changes in resting hormonal concentrations, as many studies have not shown a significant change during resistance training despite increases in muscle strength and hypertrophy. Anabolic hormones such as testosterone and the superfamily of growth hormones (GH) have been shown to be elevated during 15-30 minutes of post-resistance exercise providing an adequate stimulus is present. Protocols high in volume, moderate to high in intensity, using short rest intervals and stressing a large muscle mass, tend to produce the greatest acute hormonal elevations (e.g. testosterone, GH and the catabolic hormone cortisol) compared with low-volume, high-intensity protocols using long rest intervals. Other anabolic hormones such as insulin and insulin-like growth factor-1 (IGF-1) are critical to skeletal muscle growth. Insulin is regulated by blood glucose and amino acid levels. However, circulating IGF-1 elevations have been reported following resistance exercise presumably in response to GH stimulated hepatic secretion. Recent evidence indicates that muscle isoforms of IGF-1 may play a substantial role in tissue remodelling via up-regulation by mechanical signalling (i.e. increased gene expression resulting from stretch and tension to the muscle cytoskeleton leading to greater protein synthesis rates). Acute elevations in catecholamines are critical to optimal force production and energy liberation during resistance exercise. More recent research has shown the importance of acute hormonal elevations and mechanical stimuli for subsequent up- and down-regulation of cytoplasmic steroid receptors needed to mediate the hormonal effects. Other factors such as nutrition, overtraining, detraining and circadian patterns of hormone secretion are critical to examining the hormonal responses and adaptations to resistance training. PMID- 15831066 TI - Spatial profiles of local and nonlocal effects upon contrast detection/discrimination from classification images. AB - We used classification images (A. J. Ahumada, Jr., & J. Lovell, 1971) to estimate the perceptual filter in a task designed to assess both local and nonlocal effects upon contrast detection/discrimination. Three observers performed a yes/no detection or discrimination task of a uniform circular decrement (radius = 0.68 deg) near threshold presented for 100 to 400 ms. Stimuli were presented in ring image noise that either covered the signal and an annular surrounding area (out to 1.36 deg), or only the surrounding annular area (out to 1.36 deg). Both the signal and the annular surround appeared on a uniform background. With ring noise over both the signal and surround, the amplitudes of the classification images in the signal area decreased as radial distance increased from the signal/surround border, and no effect of the surround was found. With ring noise only in the surround, classification images indicated noncontiguous effects at both the signal/surround border (local) and the surround/background border (nonlocal). The spatial extents of the nonlocal effects (< 0.07 deg) were smaller than local effects (0.25 deg), whereas the peak amplitudes of the local and nonlocal effects were comparable. These results suggest that the nonlocal effects were smaller than the local effects, and that the smaller effects would be due to smaller effective areas, as opposed to smaller amplitudes over the same area. Little or no change was found in the classification images across stimulus duration, suggesting that both the local and nonlocal processes found in this study were completed within 100 ms. PMID- 15831067 TI - Are faces processed like words? A diagnostic test for recognition by parts. AB - Do we identify an object as a whole or by its parts? This simple question has been surprisingly hard to answer. It has been suggested that faces are recognized as wholes and words are recognized by parts. Here we answer the question by applying a test for crowding. In crowding, a target is harder to identify in the presence of nearby flankers. Previous work has described crowding between objects. We show that crowding also occurs between the parts of an object. Such internal crowding severely impairs perception, identification, and fMRI face-area activation. We apply a diagnostic test for crowding to a word and a face, and we find that the critical spacing of the parts required for recognition is proportional to distance from fixation and independent of size and kind. The critical spacing defines an isolation field around the target. Some objects can be recognized only when each part is isolated from the rest of the object by the critical spacing. In that case, recognition is by parts. Recognition is holistic if the observer can recognize the object even when the whole object fits within a critical spacing. Such an object has only one part. Multiple parts within an isolation field will crowd each other and spoil recognition. To assess the robustness of the crowding test, we manipulated familiarity through inversion and the face- and word-superiority effects. We find that threshold contrast for word and face identification is the product of two factors: familiarity and crowding. Familiarity increases sensitivity by a factor of x1.5, independent of eccentricity, while crowding attenuates sensitivity more and more as eccentricity increases. Our findings show that observers process words and faces in much the same way: The effects of familiarity and crowding do not distinguish between them. Words and faces are both recognized by parts, and their parts -- letters and facial features -- are recognized holistically. We propose that internal crowding be taken as the signature of recognition by parts. PMID- 15831068 TI - Stereo-slant adaptation is high level and does not involve disparity coding. AB - We have investigated the potential stages of visual processing at which adaptation may occur to a slanted surface produced by horizontal magnification. Predictions of three hypotheses were tested utilizing a property of depth from binocular disparity, namely that slant scales with distance. If adaptation occurs at the disparity level, then the after-effect expressed in units of horizontal magnification will be independent of the test distance. If adaptation occurs at either a perceived slant or mapping level, then the after-effect, expressed in units of slant, will be independent of the test distance. If adaptation is contingent on distance, then the after-effect will not transfer over distance. Subjects adapted to a stereo-defined slanted surface at a distance of 57 cm. The after-effect was measured with a test stimulus at a distance of 28, 57, 85, or 114 cm by means of a nulling method. When the after-effect was expressed in units of slant, we found that it was larger at the adapting distance than other test distances, and that the after-effect was constant at test distances different from the adaptation distance. These results suggest that two types of adaptation occurred, namely adaptation on a mapping/perception level and adaptation contingent on distance. PMID- 15831069 TI - Setting up the target template in visual search. AB - Top-down knowledge about the target is essential in visual search. It biases visual attention to information that matches the target-defining criteria. Extensive research in the past has examined visual search when the target is defined by fixed criteria throughout the experiment, with few studies investigating how subjects set up the target. To address this issue, we conducted five experiments using random polygons and real-world objects, allowing the target criteria to change from trial to trial. On each trial, subjects first see a cue informing them about the target, followed 200-1000 ms later by the search array. We find that when the cue matches the target exactly, search speed increases and the slope of response time-set size function decreases. Deviations from the exact match in size or orientation slow down search speed, although they lead to faster speed compared with a neutral cue or a semantic cue. We conclude that the template set-up process uses detailed visual information, rather than schematic or semantic information, to find the target. PMID- 15831070 TI - Independent anatomical and functional measures of the V1/V2 boundary in human visual cortex. AB - The cerebral cortex has both anatomical and functional specialization, but the level of correspondence between the two in the human brain has remained largely elusive. Recent successes in high-resolution magnetic resonance imaging of myeloarchitecture patterns in the cortex suggest that it may now be possible to compare directly human anatomy and function in vivo. We independently investigated the anatomical and functional borders between primary and secondary human visual areas (V1 and V2) in vivo. Functional borders were mapped with functional magnetic resonance imaging (fMRI) using a narrow, vertical black and white contrast-reversing wedge. In three separate scanning sessions, anatomical images were collected at three different slice orientations (300 microm x 300 microm, slice thickness, 1.5 mm). The anatomical signature of V1 was determined by the presence of a hypointense band in the middle of the cortical gray matter. The band was identified in between 81% and 33% (mean 57%) of V1 defined using fMRI, and less than 5% of the identified band was in cortex outside V1. Intensity profiles taken through the gray matter on the V1 and V2 sides of the functional border indicate a measurable difference in the size of the hypointense band for all subjects. This is the first demonstration that the definition of V1 by fMRI closely matches the anatomically defined striate cortex in the human brain. The development of very high-resolution structural MRI may permit the definition of cortical areas based on myeloarchitecture when functional definition is not possible. PMID- 15831071 TI - Reaching for visual cues to depth: the brain combines depth cues differently for motor control and perception. AB - Vision provides a number of cues about the three-dimensional (3D) layout of objects in a scene that could be used for planning and controlling goal-directed behaviors such as pointing, grasping, and placing objects. An emerging consensus from the perceptual work is that the visual brain is a near-optimal Bayesian estimator of object properties, for example, by integrating cues in a way that accounts for differences in their reliability. We measured how the visuomotor system integrates binocular and monocular cues to 3D surface orientation to guide the placement of objects on a slanted surface. Subjects showed qualitatively similar results to those found in perceptual studies--they gave more weight to binocular cues at low slants and more weight to monocular cues like texture at high slants. We compared subjects' performance in the visuomotor task with their performance on matched perceptual tasks that required an observer to estimate the same 3D surface properties needed to control the motor behavior. The relative influence of binocular and monocular cues changed in qualitatively the same way across stimulus conditions in the two types of task; however, subjects gave significantly more weight to binocular cues for controlling hand movements than for making explicit perceptual judgments in these tasks. Thus, the brain changes how it integrates visual cues based not only on the information content of stimuli, but also on the task for which the information is used. PMID- 15831072 TI - Experience-expectant development of contour integration mechanisms in human visual cortex. AB - Extended contours are a common feature of natural images. Most previous studies have considered contour integration as a two-dimensional process of linking like oriented elements along their common orientation axis. Yet contours exist in a three-dimensional world, and one might therefore ask about the relationship between contour integration and binocular vision. Using an event-related potential assay of contour integration, we demonstrate that patients with strabismic amblyopia show a relative insensitivity to Gabor-defined contours in their dominant eyes, all of which had normal acuity. These deficits were not seen in the dominant eyes of patients with anisometropic amblyopia without strabismus, but were present in the amblyopic eyes of patients with either strabismus or anisometropia. Deficits were also found in both eyes of strabismus patients who had normal visual acuity in each eye, but who had strongly reduced or absent stereopsis. These results suggest that the maturation of contour detection mechanisms depends at least in part on the presence of normal binocular interaction during a developmental critical period. PMID- 15831073 TI - Reference frames in early motion detection. AB - To perceive the real motion of objects in the world while moving the eyes, retinal motion signals must be compensated by information about eye movements. Here we study when this compensation takes place in the course of visual processing, and whether uncompensated motion signals are ever available. We used a paradigm based on asymmetry in motion detection: Fast-moving objects are found easier among slow-moving distractors than are slow objects among fast distractors. By coupling object motion to eye motion, we created stimuli that moved fast on the retina but slowly in an eye-independent reference frame, or vice versa. In the 100 ms after stimulus onset, motion detection is dominated by retinal motion, uncompensated for eye movements. As early as 130 ms, compensated signals become available: objects that move slowly on the retina but fast in an eye-independent frame are detected as easily as those that move fast on the retina. PMID- 15831074 TI - Classification images predict absolute efficiency. AB - How well do classification images characterize human observers' strategies in perceptual tasks? We show mathematically that from the classification image of a noisy linear observer, it is possible to recover the observer's absolute efficiency. If we could similarly predict human observers' performance from their classification images, this would suggest that the linear model that underlies use of the classification image method is adequate over the small range of stimuli typically encountered in a classification image experiment, and that a classification image captures most important aspects of human observers' performance over this range. In a contrast discrimination task and in a shape discrimination task, we found that observers' absolute efficiencies were generally well predicted by their classification images, although consistently slightly (approximately 13%) higher than predicted. We consider whether a number of plausible nonlinearities can account for the slight under prediction, and of these we find that only a form of phase uncertainty can account for the discrepancy. PMID- 15831075 TI - Osteoclastogenesis: the role of calcium and calmodulin. AB - Enhanced osteoclastogenesis is an important pathological feature in several aging associated bone diseases. Thus, research activities on osteoclastogenesis have been intense during the last ten years. There has been great progress made in this field, however, and in this review, we will focus on current advances in understanding the role of Ca2+/calmodulin signaling in osteoclastogenesis. There are two major Ca2+/calmodulin signaling pathways emerging as important in osteoclastogenesis. The first is from our recent data, which has established a specific role for calmodulin in osteoclastogenesis and, more specifically, calmodulin-dependent kinase II (CaMKII). The other is that a pathway involving RANK-Ca2+-calmodulin-calcineurin-NFAT is critical for osteoclastogenesis. Collectively, these reports highlight the importance of Ca2+/calmodulin signaling in osteoclastogenesis, which may present novel targets for the new therapeutic agents to combat bone loss. PMID- 15831076 TI - Parathyroid hormone-related protein in prostate cancer. AB - Although originally discovered as the peptide responsible for humoral hypercalcemia of malignancy (HHM), parathyroid hormone-related protein (PTHrP) has been shown to play a major role in fetal development. In the adult, it is widely distributed in normal and various cancer tissues. In spite of the rarity of HHM in prostate cancer, PTHrP is widely distributed in prostate cancer cells. PTHrP is a precursor molecule with generation of various fragments with distinct biological activities. More recent studies have shown that there is intranuclear localization of PTHrP and that intracrine effects of the peptide are involved in promoting processes that result in tumor progression (nall proliferation, apoptosis, cell attachment and angiogenesis) in prostate cancer. PTHrP expression is controlled by three distinct promoters, with P3 being used most often in cancer cells. The factors that control PTHrP production via interaction with the promoters are growth factors, androgens, vitamin D analogs, and adenoviral proteins. TGF-beta and its effector Smad3 activate the P3 promoter through an AGAC box and an Ets binding site involving Ets1 and to some extent Ets2 proteins. In addition, TGF-beta stimulates P3 promoter activity via Smad-independent pathways that involve the p38 MAP kinase. Although the addition of PTHrP or transfection with the PTHrP gene in prostate cells results in effects that promote tumor development, studies that employ inhibition of PTHrP activity in vitro and in vivo are needed to establish a definitive role of this peptide in the pathogenesis of prostate cancer. PMID- 15831077 TI - Heparan sulfate proteoglycans: key players in cartilage biology. AB - The extracellular matrix (ECM) plays a fundamental role in skeletal patterning and formation of the vertebrate skeleton. This review focuses on the fundamental roles associated with heparan sulfate (HS) proteoglycans in the ECM during cartilage development, which include regulation of gene expression, presentation of growth factors, establishment of morphogen gradients, and modulation of blood homeostasis. The importance of enzymes involved in biosynthesis and assembly of heparan sulfate is also discussed. Finally, the current evidence for functions of individual HS proteoglycans and biosynthetic enzymes based upon human genetic mutation associations with disease and genetic manipulation in transgenic mice is presented. These findings highlight the important role played by HS proteoglycans, such as perlecan, in cartilage development and skeletal growth. PMID- 15831078 TI - Nuclear transport of steroid hormone receptors. AB - Nuclear receptors (NRs) comprise a superfamily of ligand-regulated transcription factors implicated in a host of physiological processes, including development, differentiation, and proliferation. Translated in the cytoplasm, NRs must undergo import into the nucleus in order to modulate transcription of target genes in response to cognate hormones. NRs also undergo export from the nucleus, and there is emerging evidence that NR nucleocytoplasmic shuttling contributes to their regulation. Nucleocytoplasmic shuttling may provide a nexus for crosstalk between NRs and kinase pathways. Here we review some of the key studies on nuclear import and export of steroid hormone receptors within the NR superfamily, address some of the challenges in experimental dissection of NR transport and discuss recent findings linking specific kinase pathways to NR export. PMID- 15831079 TI - Apoptosis in membranous bone formation: role of fibroblast growth factor and bone morphogenetic protein signaling. AB - Membranous ossification occurs by the condensation of mesenchymal cells followed by their progressive differentiation into osteoblasts that form a mineralized matrix in ossification centers. The balance between proliferating and differentiated osteogenic cells at the suture areas between calvarial bones is essential for the control of suture maintenance and membranous bone formation. The mechanisms of regulation of na apoptosis in suture areas begin to be understood. Fibroblast growth factor (FGF) and bone morphogenetic protein (BMP) are important regulators of mesenchymal, preosteoblast, and osteoblast apoptosis in suture areas. Perturbations in FGF or BMP signaling lead to alter the number of apoptotic osteogenic cells, resulting in premature or delayed suture closure. Recent data indicate that FGF signaling downregulates preosteoblast apoptosis, thereby preventing premature fusion of adjacent mineralizing extremities. In contrast, continuous FGF signaling or constitutive FGF receptor activation, as well as BMP signaling, upregulate osteoblast apoptosis. Additionally, multiple signaling mechanisms, including PI3K and PKC, appear to be involved in the control of calvarial osteoblast apoptosis by FGF and BMP. These mechanisms allow a fine control of the number of functional bone-forming cells and, thereby, the normal progression of membranous bone formation. PMID- 15831081 TI - Inhibitory effect(s) of polymeric black tea polyphenols on the formation of B(a)P derived DNA adducts in mouse skin. AB - The biological activities and chemopreventive properties of green tea polyphenols have been demonstrated, while similar information regarding newly formed major polymeric polyphenols in black tea are not available. Cancer chemoprevention may be achieved by the inhibition of any stage of carcinogenesis. In the present study, we investigated the anti-initiating effects of five polymeric black tea polyphenol (PBP) fractions, by determining their effects on the formation of [3H] B(a)P-derived DNA adducts as well as the activity of cytochrome P-450 isozymes CYP 1A1 and 1A2 in vitro employing rat liver microsomes. PBP 1-3 inhibited both the microsome catalyzed [3H]-B(a)P-derived DNA adduct formation as well as the activity of CYP 1A1 and 1A2 as assessed by the decreased formation of resorufin from the respective substrates. Further investigation revealed that topical pretreatment(s) of mice with PBP 1-5 (200 mug/day x 4) resulted in a significant decrease in the levels of single topical B(a)P (1 mg/mouse) - induced DNA adducts in epidermal DNA determined by employing 32P-post labeling analysis. Overall, our results suggest that black tea-derived PBPs have one of the chemopreventive properties shown by monomeric green tea polyphenols. PMID- 15831082 TI - Black tea protects thymocytes in tumor-bearing animals by differential regulation of intracellular ROS in tumor cells and thymocytes. AB - The accumulated in vitro evidence indicates that many tumors induce T-cell apoptosis as a mechanism of inhibiting antitumor activity. This downregulation of cell-mediated immune functions occurring at the late stages of the disease may be causally related to the thymic involution, because the thymus is the major site of T-cell maturation, extensive proliferation, and differentiation. Our results showed that in Erhlich's ascites carcinoma cell (EAC)-bearing mice, the number of EAC was inversely proportional to the thymocyte count in the host's thymus, which is the primary immune organ. Further studies indicated the presence of tumor induced thymocyte apoptosis in EAC bearers. Black tea prolonged the survival of the tumor bearer by successfully restricting tumor progression as well as protecting the thymus from tumor insult. In fact, black tea inhibited thymic apoptosis while inducing programmed cell death of EAC. Interestingly, the tea regulated the oxidant status differentially in EAC and thymocytes--i.e., it reduced the EAC-induced reactive oxygen species (ROS) generation in the thymus while activating the same in the EAC. A similar effect of black tea was obtained when thymocytes were cultured in the presence of cell-free ascitic fluid, thereby indicating that black tea could directly reduce oxidative stress, an activity independent of its tumoricidal property. As a result, the maturation block in thymocyte subpopulations in tumor bearers was ameliorated significantly in black tea-treated animals. Our results demonstrate that black tea protects thymocytes in the tumor bearer by regulating the intracellular ROS in tumor cells and thymocytes differentially, thereby strengthening its candidacy in future anticancer therapeutic regimens. PMID- 15831083 TI - Antioxidant potential of black tea against 7,12-dimethylbenz(a)anthracene- induced oxidative stress in Swiss albino mice. AB - Oxygen free radicals and related reactive species have been implicated in the etiology of many diseases, such as atherosclerosis, neurodegenerative disorders, and cancer. Antioxidant enzymes exist in cells to protect against the effects of these free radicals and other oxygen-derived species, which are produced during the oxidative stress. Tea (Camellia sinensis) is the most commonly consumed beverage worldwide. Both green and black tea are known to possess many pharmacological properties, including antioxidant, antipyretic, antibacterial, and antineoplastic effects. In the present study, the preventive effects of black tea extract (BTE) was evaluated in Swiss albino mice against 7,12-dimethyl benz(a)anthracene (DMBA)-induced oxidative stress. The animals were given 0.5%, 1%, and 1.5% BTE as the sole source of drinking solution for 1 week prior to the administration of DMBA, which was given orally as a single dose of 50 mg/kg body weight. At the end of the study period, the liver, kidney, and prostate tissues were dissected out for the determination of antioxidant enzyme levels (catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase), and lipid peroxidation. A dose-dependent protective effect of BTE against DMBA induced depletion in enzymes activity was observed in all three tissues examined. Similarly, a significant dose-dependent inhibition of the lipid peroxidation caused by DMBA was observed in the BTE-administered animals in all three tissues examined. Our results revealed that BTE provides protection against oxidative damage induced by xenobiotics. PMID- 15831084 TI - Correlation of apoptosis with comet formation induced by tea polyphenols in human leukemia cells. AB - Induction of apoptosis is an important approach to cancer control. Apart from morphological changes in cells, apoptosis is characterized by fragmentation of nuclear DNA. The characteristic DNA ladder formation that is observed on gel electrophoresis does not reflect the DNA breakdown in individual cells; contributions from small subpopulations are usually overlooked. On the other hand, alkaline comet assay as measured by single cell gel electrophoresis accurately measures DNA fragmentation at a single cell level. The comet assay was originally developed as a cytogenetic test to measure the genotoxicity of various chemicals. However, the comet image generated by an apoptotic cell is different from that obtained with a cell treated for a short time with a genotoxic agent. In the present study using human leukemic cells, typical apoptotic features such as morphological characteristics, FACS analysis, caspase activation, and expression of apoptosis-related genes as induced by tea polyphenols have been found to correlate with the comet tail formation. It is apparent from the high degree of correlation observed between the comet tail moment and each parameter of apoptosis that the comet assay can accurately reflect the measure of DNA fragmentation and, hence, can be used to detect a cell undergoing apoptosis. PMID- 15831085 TI - Amelioration of sodium arsenite-induced clastogenicity by tea extracts in Chinese hamster v79 cells. AB - Since the early 1980s, an alarming problem of groundwater arsenic (As) contamination has devastated many districts of West Bengal in India. People drinking As-contaminated water have been suffering severe health problems such as hyperkeratosis, blackfoot disease, neuropathy, and cancer of various sites. DNA damage and genetic instability induced by the inorganic arsenicals present in water are thought to be prerequisites for the initiation of carcinogenesis. Many natural polyphenols, which are consumed through our daily diet, possess excellent cancer chemopreventive properties. Tea, a popular beverage worldwide and rich in polyphenols, has exhibited many health benefits. The present study was conducted to examine the anticlastogenic action of tea extracts (both green and black) against the As-induced chromosomal aberrations. We also evaluated the role of tea in inducing antioxidant enzymes such as superoxide dismutase and catalase to provide protection against the oxidative stress induced by As. Our results demonstrated that tea extracts, particularly Darjeeling tea extract, are effective in counteracting the clastogenicity (chromatid breaks, in particular) of the most potent form of As, sodium arsenite. The antioxidant function of tea in reducing clastogenicity may be partly due to the induction of phase II detoxification enymes, such as superoxide dismutase and catalase. Our results suggest that the use of tea may be an effective approach in combating the health crisis generated by As. PMID- 15831086 TI - Black tea (Camellia sinensis) as a chemopreventive agent in oral precancerous lesions. AB - Oral carcinoma is the most common malignancy found in adult Indian men and the third most common in adult Indian women. About half of all cases are found to be associated with precancerous lesions, chiefly leukoplakia. We wanted to explore the possible benefits of black tea (Camellia sinensis) administered to patients with oral leukoplakia. Eighty-two subjects with oral leukoplakia underwent micronuclei and chromosomal assays on exfoliated oral mucosal epithelium, after which they received black tea in a fixed regimen. The micronuclei assay was repeated at 6 months, and the chromosomal study at 1 year. After the first year, the first 15 patients entered onto this study showed a significant decrease in the micronuclei frequency and chromosomal aberrations, which correlated with the clinical improvement. Several in vitro and animal studies have suggested the efficacy of tea in the chemoprevention of cancer. To the best of our knowledge, this is the first report on the effect of black tea in oral leukoplakia. PMID- 15831087 TI - The molecular make-up of a tumour: proteomics in cancer research. AB - The enormous progress in proteomics, enabled by recent advances in MS (mass spectrometry), has brought protein analysis back into the limelight of cancer research, reviving old areas as well as opening new fields of study. In this review, we discuss the basic features of proteomic technologies, including the basics of MS, and we consider the main current applications and challenges of proteomics in cancer research, including (i) protein expression profiling of tumours, tumour fluids and tumour cells; (ii) protein microarrays; (iii) mapping of cancer signalling pathways; (iv) pharmacoproteomics; (v) biomarkers for diagnosis, staging and monitoring of the disease and therapeutic response; and (vi) the immune response to cancer. All these applications continue to benefit from further technological advances, such as the development of quantitative proteomics methods, high-resolution, high-speed and high-sensitivity MS, functional protein assays, and advanced bioinformatics for data handling and interpretation. A major challenge will be the integration of proteomics with genomics and metabolomics data and their functional interpretation in conjunction with clinical results and epidemiology. PMID- 15831088 TI - Analysis of human neurological disorders using mutagenesis in the mouse. AB - The mouse continues to play a vital role in the deciphering of mammalian gene function and the modelling of human neurological disease. Advances in gene targeting technologies have facilitated the efficiency of generating new mouse mutants, although this valuable resource has rapidly expanded in recent years due to a number of major random mutagenesis programmes. The phenotype-driven mutagenesis screen at the MRC Mammalian Genetics Unit has generated a significant number of mice with potential neurological defects, and our aim has been to characterize selected mutants on a pathological and molecular level. Four lines are discussed, one displaying late-onset ataxia caused by Purkinje cell loss and an allelic series of three tremor mutants suffering from hypomyelination of the peripheral nerve. Molecular analysis of the causative mutation in each case has provided new insights into functional aspects of the mutated proteins, illustrating the power of mutagenesis screens to generate both novel and clinically relevant disease models. PMID- 15831089 TI - 5-HT in systemic hypertension: foe, friend or fantasy? AB - Since its discovery by Erspamer in the 1930s and identification by Page in the 1950s, 5-HT (5-hydroxytryptamine; serotonin) has been an elusive candidate as a substance that plays a role in the disease of high blood pressure, also known as hypertension. In both animal and human hypertension, arterial contraction to 5-HT is profoundly enhanced. Additionally, 5-HT is a vascular smooth muscle cell mitogen. Because both increased arterial contractility and smooth muscle growth contribute to the disease of hypertension, it is logical to believe that 5-HT is a potential cause of disease, and thus a foe. However, decades of research have produced conflicting results as to the potential role of 5-HT in hypertension. This review will discuss historical findings which both support and refute the involvement of 5-HT in hypertension, and pose some new questions that may reveal novel ways for 5-HT to modify vascular control of blood pressure. PMID- 15831090 TI - Redox regulation of neutrophil apoptosis and the systemic inflammatory response syndrome. AB - SIRS (systemic inflammatory response syndrome) may result from a wide variety of non-infective insults. Surgery is a recognized cause of SIRS, the onset of which can have adverse prognostic significance. Neutrophil activation is a key histopathological feature of SIRS, and neutrophil clearance through programmed cell death or apoptosis is an essential step in its resolution. Increasingly, it is recognized that ROS (reactive oxygen species), such as those generated by activated neutrophils during cardiac surgery, may have a regulatory role, influencing neutrophil lifespan and thus inflammation. In this review, we discuss the continuing importance of SIRS as a herald of inflammation and the role of neutrophil longevity in the resolution of inflammation, and we consider recent evidence for the regulation of neutrophil apoptosis by ROS. PMID- 15831091 TI - CD34+ cells cultured in stem cell factor and interleukin-2 generate CD56+ cells with antiproliferative effects on tumor cell lines. AB - In vitro stimulation of CD34+ cells with IL-2 induces NK cell differentiation. In order to define the stages of NK cell development, which influence their generation from CD34 cells, we cultured G-CSF mobilized peripheral blood CD34+ cells in the presence of stem cell factor and IL-2. After three weeks culture we found a diversity of CD56+ subsets which possessed granzyme A, but lacked the cytotoxic apparatus required for classical NK-like cytotoxicity. However, these CD56+ cells had the unusual property of inhibiting proliferation of K562 and P815 cell lines in a cell-contact dependent fashion. PMID- 15831092 TI - Occupational risk of tuberculosis transmission in a low incidence area. AB - BACKGROUND: To investigate the occupational risk of tuberculosis (TB) infection in a low-incidence setting, data from a prospective study of patients with culture-confirmed TB conducted in Hamburg, Germany, from 1997 to 2002 were evaluated. METHODS: M. tuberculosis isolates were genotyped by IS6110 RFLP analysis. Results of contact tracing and additional patient interviews were used for further epidemiological analyses. RESULTS: Out of 848 cases included in the cluster analysis, 286 (33.7%) were classified into 76 clusters comprising 2 to 39 patients. In total, two patients in the non-cluster and eight patients in the cluster group were health-care workers. Logistic regression analysis confirmed work in the health-care sector as the strongest predictor for clustering (OR 17.9). However, only two of the eight transmission links among the eight clusters involving health-care workers had been detected previously. Overall, conventional contact tracing performed before genotyping had identified only 26 (25.2%) of the 103 contact persons with the disease among the clustered cases whose transmission links were epidemiologically verified. CONCLUSION: Recent transmission was found to be strongly associated with health-care work in a setting with low incidence of TB. Conventional contact tracing alone was shown to be insufficient to discover recent transmission chains. The data presented also indicate the need for establishing improved TB control strategies in health-care settings. PMID- 15831093 TI - +Psychometric evaluation of the MacDQoL individualised measure of the impact of macular degeneration on quality of life. AB - BACKGROUND: The MacDQoL is an individualised measure of the impact of macular degeneration (MD) on quality of life (QoL). There is preliminary evidence of its psychometric properties and sensitivity to severity of MD. The aim of this study was to carry out further psychometric evaluation with a larger sample and investigate the measure's sensitivity to MD severity. METHODS: Patients with MD (n = 156: 99 women, 57 men, mean age 79 +/- 13 years), recruited from eye clinics (one NHS, one private) completed the MacDQoL by telephone interview and later underwent a clinic vision assessment including near and distance visual acuity (VA), comfortable near VA, contrast sensitivity, colour recognition, recovery from glare and presence or absence of distortion or scotoma in the central 10 degrees of the visual field. RESULTS: The completion rate for the MacDQoL items was 99.8%. Of the 26 items, three were dropped from the measure due to redundancy. A fourth was retained in the questionnaire but excluded when computing the scale score. Principal components analysis and Cronbach's alpha (0.944) supported combining the remaining 22 items in a single scale. Lower MacDQoL scores, indicating more negative impact of MD on QoL, were associated with poorer distance VA (better eye r = -0.431 p < 0.001; worse eye r = -0.350 p < 0.001; binocular vision r = -0.419 p < 0.001) and near VA (better eye r = 0.326 p < 0.001; worse eye r = -0.226 p < 0.001; binocular vision r = -0.326 p < 0.001). Poorer MacDQoL scores were associated with poorer contrast sensitivity (better eye r = 0.392 p < 0.001; binocular vision r = 0.423 p < 0.001), poorer colour recognition (r = 0.417 p < 0.001) and poorer comfortable near VA (r = 0.283, p < 0.001). The MacDQoL differentiated between those with and without binocular scotoma (U = 1244 p < 0.001). CONCLUSION: The MacDQoL 22-item scale has excellent internal consistency reliability and a single-factor structure. The measure is acceptable to respondents and the generic QoL item, MD-specific QoL item and average weighted impact score are related to several measures of vision. The MacDQoL demonstrates that MD has considerable negative impact on many aspects of QoL, particularly independence, leisure activities, dealing with personal affairs and mobility. The measure may be valuable for use in clinical trials and routine clinical care. PMID- 15831094 TI - Automated measurement of cell motility and proliferation. AB - BACKGROUND: Time-lapse microscopic imaging provides a powerful approach for following changes in cell phenotype over time. Visible responses of whole cells can yield insight into functional changes that underlie physiological processes in health and disease. For example, features of cell motility accompany molecular changes that are central to the immune response, to carcinogenesis and metastasis, to wound healing and tissue regeneration, and to the myriad developmental processes that generate an organism. Previously reported image processing methods for motility analysis required custom viewing devices and manual interactions that may introduce bias, that slow throughput, and that constrain the scope of experiments in terms of the number of treatment variables, time period of observation, replication and statistical options. Here we describe a fully automated system in which images are acquired 24/7 from 384 well plates and are automatically processed to yield high-content motility and morphological data. RESULTS: We have applied this technology to study the effects of different extracellular matrix compounds on human osteoblast-like cell lines to explore functional changes that may underlie processes involved in bone formation and maintenance. We show dose-response and kinetic data for induction of increased motility by laminin and collagen type I without significant effects on growth rate. Differential motility response was evident within 4 hours of plating cells; long-term responses differed depending upon cell type and surface coating. Average velocities were increased approximately 0.1 microm/min by ten-fold increases in laminin coating concentration in some cases. Comparison with manual tracking demonstrated the accuracy of the automated method and highlighted the comparative imprecision of human tracking for analysis of cell motility data. Quality statistics are reported that associate with stage noise, interference by non-cell objects, and uncertainty in the outlining and positioning of cells by automated image analysis. Exponential growth, as monitored by total cell area, did not linearly correlate with absolute cell number, but proved valuable for selection of reliable tracking data and for disclosing between-experiment variations in cell growth. CONCLUSION: These results demonstrate the applicability of a system that uses fully automated image acquisition and analysis to study cell motility and growth. Cellular motility response is determined in an unbiased and comparatively high throughput manner. Abundant ancillary data provide opportunities for uniform filtering according to criteria that select for biological relevance and for providing insight into features of system performance. Data quality measures have been developed that can serve as a basis for the design and quality control of experiments that are facilitated by automation and the 384 well plate format. This system is applicable to large scale studies such as drug screening and research into effects of complex combinations of factors and matrices on cell phenotype. PMID- 15831095 TI - Subfunctionalization of duplicated genes as a transition state to neofunctionalization. AB - BACKGROUND: Gene duplication has been suggested to be an important process in the generation of evolutionary novelty. Neofunctionalization, as an adaptive process where one copy mutates into a function that was not present in the pre duplication gene, is one mechanism that can lead to the retention of both copies. More recently, subfunctionalization, as a neutral process where the two copies partition the ancestral function, has been proposed as an alternative mechanism driving duplicate gene retention in organisms with small effective population sizes. The relative importance of these two processes is unclear. RESULTS: A set of lattice model genes that fold and bind to two peptide ligands with overlapping binding pockets, but not a third ligand present in the cell was designed. Each gene was duplicated in a model haploid species with a small constant population size and no recombination. One set of models allowed subfunctionalization of binding events following duplication, while another set did not allow subfunctionalization. Modeling under such conditions suggests that subfunctionalization plays an important role, but as a transition state to neofunctionalization rather than as a terminal fate of duplicated genes. There is no apparent selective pressure to maintain redundancy. CONCLUSION: Subfunctionalization results in an increase in the preservation of duplicated gene copies, including those that are neofunctionalized, but never represents a substantial fraction of duplicate gene copies at any evolutionary time point and ultimately leads to neofunctionalization of those preserved copies. This conclusion also may reflect changes in gene function after duplication with time in real genomes. PMID- 15831096 TI - A balanced review of the status T cell-based therapy against cancer. AB - A recent commentary stirred intense controversy over the status of anti-cancer immunotherapy. The commentary suggested moving beyond current anti-cancer vaccines since active-specific immunization failed to match expectations toward a more aggressive approach involving the adoptive transfer of in vitro expanded tumor antigen-specific T cells. Although the same authors clarified their position in response to others' rebuttal more discussion needs to be devoted to the current status of T cell-based anti-cancer therapy. The accompanying publications review the status of adoptive transfer of cancer vaccines on one hand and active-specific immunization on the other. Hopefully, reading these articles will offer a balanced view of the current status of antigen-specific ant cancer therapies and suggest future strategies to foster unified efforts to complement either approach with the other according to specific biological principles. PMID- 15831097 TI - Immune function biomarkers in children exposed to lead and organochlorine compounds: a cross-sectional study. AB - BACKGROUND: Different organochlorines and lead (Pb) have been shown to have immunomodulating properties. Children are at greater risk for exposure to these environmental toxicants, but very little data exist on simultaneous exposures to these substances. METHODS: We investigated whether the organochlorine compounds (OC) dichlorodiphenylethylene (DDE), hexachlorobenzene (HCB), hexachlorocyclohexane (gamma-HCH), the sum of polychlorinated biphenyls (SigmaPCBs) and Pb were associated with immune markers such as immunoglobulin (Ig) levels, white blood cell (WBC), counts of lymphocytes; eosinophils and their eosinophilic granula as well as IgE count on basophils. The investigation was part of a cross-sectional environmental study in Hesse, Germany. In 1995, exposure to OC and Pb were determined, questionnaire data collected and immune markers quantified in 331 children. For the analyses, exposure (OC and Pb) concentrations were grouped in quartiles (gamma-HCH into tertiles). Using linear regression, controlling for age, gender, passive smoking, serum lipids, and infections in the previous 12 months, we assessed the association between exposures and immune markers. Adjusted geometric means are provided for the different exposure levels. RESULTS: Geometric means were: DDE 0.32 microg/L, SigmaPCBs 0.50 microg/L, HCB 0.22 microg/L, gamma-HCH 0.02 microg/L and Pb 26.8 microg/L. The SigmaPCBs was significantly associated with increased IgM levels, whereas HCB was inversely related to IgM. There was a higher number of NK cells (CD56+) with increased gamma-HCH concentrations. At higher lead concentrations we saw increased IgE levels. DDE showed the most associations with significant increases in WBC count, in IgE count on basophils, IgE, IgG, and IgA levels. DDE was also found to significantly decrease eosinophilic granula content. CONCLUSION: Low-level exposures to OC and lead (Pb) in children may have immunomodulating effects. The increased IgE levels, IgE count on basophils, and the reduction of eosinophilic granula at higher DDE concentrations showed a most consistent pattern, which could be of clinical importance in the etiology of allergic diseases. PMID- 15831098 TI - Comparison of SPECT bone scintigraphy with MRI for diagnosis of meniscal tears. AB - BACKGROUND: Scintigraphy has been considered as competitive to MRI, but limited data are available on the accuracy of single photon emission tomography (SPECT) compared with MRI for the assessment of meniscal tears. Our objective was to assess the value of SPECT in comparison to MRI. METHODS: Between January 2003 and March 2004, sixteen patients were studied with both modalities and the accuracy rates of SPECT scan results, and MRI findings in the diagnosis of meniscal tears were compared. Arthroscopy was the gold standard. RESULTS: The respective sensitivity rate, specificity rate, and positive and negative predictive accuracies of MRI were 89%, 94%, 93%, and 79% and for SPECT those were 78%, 94%, 94%, and 88%. There was good agreement on the presence or absence of tears between two modalities (kappa statistic = 0.699). CONCLUSION: SPECT and MRI are both valuable imaging techniques. SPECT is a useful alternative when MRI is unavailable or unsuitable and it is beneficial when more possible accuracy is desired (such as when MRI results are either inconclusive or conflict with other clinical data). PMID- 15831099 TI - Identifying future models for delivering genetic services: a nominal group study in primary care. AB - BACKGROUND: To enable primary care medical practitioners to generate a range of possible service delivery models for genetic counselling services and critically assess their suitability. METHODS: Modified nominal group technique using in primary care professional development workshops. RESULTS: 37 general practitioners in Wales, United Kingdom too part in the nominal group process. The practitioners who attended did not believe current systems were sufficient to meet anticipated demand for genetic services. A wide range of different service models was proposed, although no single option emerged as a clear preference. No argument was put forward for genetic assessment and counselling being central to family practice, neither was there a voice for the view that the family doctor should become skilled at advising patients about predictive genetic testing and be able to counsel patients about the wider implications of genetic testing for patients and their family members, even for areas such as common cancers. Nevertheless, all the preferred models put a high priority on providing the service in the community, and often co-located in primary care, by clinicians who had developed expertise. CONCLUSION: There is a need for a wider debate about how healthcare systems address individual concerns about genetic concerns and risk, especially given the increasing commercial marketing of genetic tests. PMID- 15831100 TI - Microtubule plus-ends reveal essential links between intracellular polarization and localized modulation of endocytosis during division-plane establishment in plant cells. AB - BACKGROUND: A key event in plant morphogenesis is the establishment of a division plane. A plant-specific microtubular preprophase band (PPB) accurately predicts the line of cell division, whereas the phragmoplast, another plant-specific array, executes cell division by maintaining this predicted line. Although establishment of these specific arrays apparently involves intracellular repolarization events that focus cellular resources to a division site, it still remains unclear how microtubules position the cell division planes. Here we study GFP-AtEB1 decorated microtubule plus-ends to dissect events at the division plane. RESULTS: Early mitotic events included guided growth of endoplasmic microtubules (EMTs) towards the PPB site and their coincident localization with endocytic vesicles. Consequently, an endosomal belt lay in close proximity to the microtubular PPB at its maturation and was maintained during spindle formation. During cytokinesis, EMTs radiated from the former spindle poles in a geometrical conformation correlating with cell-plate navigation and tilt-correction. Naphthylphtalamic acid (NPA), an inhibitor of polar auxin efflux, caused abnormal PPBs and shifted division planes. CONCLUSION: Our observations reveal a spatio temporal link between microtubules and intracellular polarization essential for localized endocytosis and precise establishment of the division plane in plants. Additionally, they implicate the growth regulator, auxin, in this important cellular event. PMID- 15831102 TI - Eco-epidemiological analysis of dengue infection during an outbreak of dengue fever, India. AB - BACKGROUND: This study was designed to find out a relationship of dengue infection with climatic factors such as rainfall, temperature and relative humidity during the dengue fever epidemic in the year 2003. Blood samples were collected from 1550 patients experiencing a febrile illness clinically consistent with dengue infection. Serological confirmation of Dengue Infection was done using Dengue Duo IgM and IgG Rapid Strip test (Pan Bio, Australia), which detected dengue-specific antibodies. Monthly data of total rainfall, temperature and relative humidity for the year 2003 was obtained from Meteorological Department of Delhi, New Delhi and retrospectively analyzed. RESULTS: Out of 1550 suspected cases, 893 cases (57.36%) were confirmed as serologically positive. The difference between numbers of serologically positive cases during different months was significant (p < 0.05). Larger proportions of serologically positive cases were observed among adults. Outbreak coincided mainly with the post monsoon period of subnormal rainfall. The difference between serologically positive cases as compared to serologically negative ones in post monsoon period was significantly higher (p < 0.001). The difference in the rainfall and temperature between three seasonal periods was significant (p < 0.05). CONCLUSION: This prospective study highlighted rain, temperature and relative humidity as the major and important climatic factors, which could alone or collectively be responsible for an outbreak. More studies in this regard could further reveal the correlation between the climatic changes and dengue outbreaks, which would help in making the strategies and plans to forecast any outbreak in future well in advance. PMID- 15831101 TI - Use of adenoviral E1A protein to analyze K18 promoter deregulation in colon carcinoma cells discloses a role for CtBP1 and BRCA1. AB - BACKGROUND: The promoter of the keratin 18 (K18) gene is 5- to 10-fold more active in tumorigenic (T-type) cell clones derived from the SW613-S human colon carcinoma cell line than in non-tumorigenic (NT-type) clones. We have reported previously that the mechanism responsible for this differential activity is acting on the minimal K18 promoter (TATA box and initiation site). This mechanism does not require the binding of a factor to a specific site on the DNA but involves the acetylation of a non-histone substrate. To get further insight into this mechanism, we investigated the effect of the adenovirus E1A protein on the activity of the K18 promoter, both in T and NT cells. RESULTS: Wild type adenovirus E1A protein and C-terminal deletion mutants inhibit the K18 promoter, specifically in T-type cells. The domain responsible for this inhibitory effect is located in the 12-25 region of the viral protein. E1A mutants that have lost this region but retain the PLDLS motif (the C-terminal binding site for CtBP1) stimulate the K18 promoter, specifically in NT cells. The inhibitory or stimulatory effects of the different E1A mutants are not dependent on a particular sequence of the promoter. An E1A N-terminal deletion mutant carrying point mutations in the PLDLS motif cannot stimulate the K18 promoter. CtBP1 interacts with CtIP, which is a known partner of BRCA1, itself a component of the RNA polymerase II holoenzyme. The stimulatory effect of two BRCA1 mutants, specifically in NT cells, implicates a tripartite BRCA1-CtIP-CtBP1 complex in the regulation of the K18 promoter. CONCLUSION: Since we have shown previously that the K18 promoter is stimulated by deacetylase inhibitors, specifically in NT cells, we conclude that the activity of the promoter is repressed in NT cells by a mechanism involving the recruitment, by a BRCA1/CtIP complex, of CtBP1 and associated deacetylases to the preinitiation complex. We propose a model depicting the mechanism responsible for the differential activity of the K18 promoter between T and NT cells of the SW613-S cell line. PMID- 15831103 TI - Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein. AB - BACKGROUND: Rice tungro bacilliform virus (RTBV) is a pararetrovirus, and a member of the family Caulimoviridae in the genus Badnavirus. RTBV has a long open reading frame that encodes a large polyprotein (P3). Pararetroviruses show similarities with retroviruses in molecular organization and replication. P3 contains a putative movement protein (MP), the capsid protein (CP), the aspartate protease (PR) and the reverse transcriptase (RT) with a ribonuclease H activity. PR is a member of the cluster of retroviral proteases and serves to proteolytically process P3. Previous work established the N- and C-terminal amino acid sequences of CP and RT, processing of RT by PR, and estimated the molecular mass of PR by western blot assays. RESULTS: A molecular mass of a protein that was associated with virions was determined by in-line HPLC electrospray ionization mass spectral analysis. Comparison with retroviral proteases amino acid sequences allowed the characterization of a putative protease domain in this protein. Structural modelling revealed strong resemblance with retroviral proteases, with overall folds surrounding the active site being well conserved. Expression in E. coli of putative domain was affected by the presence or absence of the active site in the construct. Analysis of processing of CP by PR, using pulse chase labelling experiments, demonstrated that the 37 kDa capsid protein was dependent on the presence of the protease in the constructs. CONCLUSION: The findings suggest the characterization of the RTBV protease domain. Sequence analysis, structural modelling, in vitro expression studies are evidence to consider the putative domain as being the protease domain. Analysis of expression of different peptides corresponding to various domains of P3 suggests a processing of CP by PR. This work clarifies the organization of the RTBV polyprotein, and its processing by the RTBV protease. PMID- 15831104 TI - Evaluation of POSSUM scoring system in patients with gastric cancer undergoing D2 gastrectomy. AB - BACKGROUND: Risk adjustment and stratification play an important role in quality assurance and in clinical research. The Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM) is a patient risk prediction model based on 12 patient characteristics and 6 characteristics of the surgery performed. However, because the POSSUM was developed for quality assessment in general surgical units, its performance within specific subgroups still requires evaluation. The aim of the present study was to assess the accuracy of POSSUM in predicting mortality and morbidity in patients with gastric cancer undergoing D2-gastrectomy. METHODS: 137 patients with gastric cancer undergoing gastrectomy were included in this study. Detailed, standardized risk assessments and thorough documentation of the post-operative courses were performed prospectively, and the POSSUM scores were then calculated. RESULTS: The 30- and 90-day mortality rates were 3.6% (n = 5) and 5.8% (n = 8), respectively. 65.7% (n = 90) of patients had normal postoperative courses without major complications, 14.6% (n = 20) had moderate and 13.9% (n = 19) had severe complications. The number of mortalities predicted by the POSSUM-Mortality Risk Score (R1) was double the actual number of mortalities occurring in the median and high-risk groups, and was more than eight times the actual number of mortalities occurring in the low-risk group (R1 < 20%). However, the calculated R1 predicted rather well in terms of severe morbidity or post-operative death in each risk group: in predicted low risk patients the actual occurrence rate (AR) of severe morbidity or post-operative death was 14%, for predicted medium risk patients the AR was 23%, and for predicted high risk patients the AR was 50% (p < 0.05). The POSSUM-Morbidity Risk Score (R2) overestimated the risk of morbidity. CONCLUSION: The POSSUM Score may be beneficial and can be used for assessment of the peri- and post-operative courses of patients with gastric carcinoma undergoing D2-gastrectomy. However, none of the scores examined here are useful for preoperative prediction of postoperative course. PMID- 15831105 TI - Improved profile HMM performance by assessment of critical algorithmic features in SAM and HMMER. AB - BACKGROUND: Profile hidden Markov model (HMM) techniques are among the most powerful methods for protein homology detection. Yet, the critical features for successful modelling are not fully known. In the present work we approached this by using two of the most popular HMM packages: SAM and HMMER. The programs' abilities to build models and score sequences were compared on a SCOP/Pfam based test set. The comparison was done separately for local and global HMM scoring. RESULTS: Using default settings, SAM was overall more sensitive. SAM's model estimation was superior, while HMMER's model scoring was more accurate. Critical features for model building were then analysed by comparing the two packages' algorithmic choices and parameters. The weighting between prior probabilities and multiple alignment counts held the primary explanation why SAM's model building was superior. Our analysis suggests that HMMER gives too much weight to the sequence counts. SAM's emission prior probabilities were also shown to be more sensitive. The relative sequence weighting schemes are different in the two packages but performed equivalently. CONCLUSION: SAM model estimation was more sensitive, while HMMER model scoring was more accurate. By combining the best algorithmic features from both packages the accuracy was substantially improved compared to their default performance. PMID- 15831107 TI - Parasitic zoonoses in Papua New Guinea. AB - Relatively few species of zoonotic parasites have been recorded in humans in Papua New Guinea. A greater number of potentially zoonotic species, mostly nematodes, occur in animals but are yet to be reported from humans. Protozoa is the best represented group of those infecting man, with Giardia duodenalis, Cryptosporidium parvum, Cyclospora cayetanesis, Toxoplasma gondii, Sarcocystis spp., Entamoeba polecki, Balantidium coli and, possibly, Blastocystis hominis. The only zoonotic helminths infecting humans include the trematode Paragonimus westermani, the cestodes Hymenolepis nana, H. diminuta and the sparganum larva of Spirometra erinacea, and the nematodes Trichinella papuae and Angiostrongylus cantonensis and, possibly, Ascaris suum. Other groups represented are Acanthocephala (Macracanthorhynchus hirudinaceus)), insects (Chrysomya bezziana, Cimex sp., Ctenocephalides spp.), and mites (Leptotrombidium spp. and, possibly Sarcoptes scabiei, and Demodex sp.). One leech (Phytobdella lineata) may also be considered as being zoonotic. The paucity of zoonotic parasite species can be attributed to long historical isolation of the island of New Guinea and its people, and the absence until recent times of large placental mammals other than pig and dog. Some zoonotic helminths have entered the country with recent importation of domestic animals, in spite of quarantine regulations, and a few more (two cestodes, one nematode and one tick) are poised to enter from neighbouring countries, given the opportunity. Improvement in water supplies, human hygiene and sanitation would reduce the prevalence of many of these parasites, and thorough cooking of meat would lessen the risk of infection by some others. PMID- 15831108 TI - Endoparasites of the blue whiting, Micromesistius poutassou from north-west Spain. AB - The communities of metazoan endoparasites of blue whiting, Micromesistius poutassou, in waters of north-west Spain were analysed and a geographical comparison made with other localities. Four hundred blue whiting collected in July 1999 and September 2000 were examined for parasites, excluding the head and gills. Six species were found: Anisakis simplex s.l. (L3), A. physeteris (L3), Hysterothylacium aduncum (L2 and L3), Stephanostomum lophii (metacercaria), S. pristis (adult), and Prosorhynchus crucibulum (metacercaria). The latter is a new host record, and A. physeteris is reported for the first time in blue whiting from the north-east Atlantic. Host gender was not a significant predictor of abundance of any helminth species, and host length was only weakly and positively related to the abundance of A. simplex. Infracommunities were species-poor, with 56% of fish harbouring only one parasite species, and 92% up to two species. Infracommunities were strongly dominated by A. simplex (389 fish) or S. lophii (6 fish). Fish length or gender, and the year of capture, did not affect species richness nor the degree of dominance. There were no significant pair-wise associations between species. Infracommunities were basically composed of several allogenic parasites with different life histories that converge in the blue whiting through the local food web. The parasite fauna of blue whiting in the study area was poor and distinctive compared with that of other localities in the north-east North Atlantic. These peculiarities might primarily be related to the composition of the local community of definitive hosts, although there might also be some influence of the geographical distribution of parasites. PMID- 15831106 TI - Can pulsed ultrasound increase tissue damage during ischemia? A study of the effects of ultrasound on infarcted and non-infarcted myocardium in anesthetized pigs. AB - BACKGROUND: The same mechanisms by which ultrasound enhances thrombolysis are described in connection with non-beneficial effects of ultrasound. The present safety study was therefore designed to explore effects of beneficial ultrasound characteristics on the infarcted and non-infarcted myocardium. METHODS: In an open chest porcine model (n = 17), myocardial infarction was induced by ligating a coronary diagonal branch. Pulsed ultrasound of frequency 1 MHz and intensity 0.1 W/cm2 (ISATA) was applied during one hour to both infarcted and non-infarcted myocardial tissue. These ultrasound characteristics are similar to those used in studies of ultrasound enhanced thrombolysis. Using blinded assessment technique, myocardial damage was rated according to histopathological criteria. RESULTS: Infarcted myocardium exhibited a significant increase in damage score compared to non-infarcted myocardium: 6.2 +/- 2.0 vs. 4.3 +/- 1.5 (mean +/- standard deviation), (p = 0.004). In the infarcted myocardium, ultrasound exposure yielded a further significant increase of damage scores: 8.1 +/- 1.7 vs. 6.2 +/- 2.0 (p = 0.027). CONCLUSION: Our results suggest an instantaneous additive effect on the ischemic damage in myocardial tissue when exposed to ultrasound of stated characteristics. The ultimate damage degree remains to be clarified. PMID- 15831109 TI - Macrofilaricidal and microfilaricidal effects of Neurolaena lobata, a Guatemalan medicinal plant, on Brugia pahangi. AB - Twelve extracts of 11 Guatemalan medicinal plants were initially screened in vitro for potential macrofilaricidal activity against Brugia pahangi, a lymphatic dwelling filarial worm, using concentrations from 125 to 1000 microg ml(-1) of each extract that could be dissolved in the culture medium. Of 12 extracts used, the ethanol extract of leaves of Neurolaena lobata showed the strongest activity against the motility of adult worms. Subsequently, the extract of N. lobata was extensively examined in vitro for macro- and micro-filaricidal effects using a series of concentrations of 500, 250, 100, 50 and 10 microg ml(-1). The effects were assessed by worm motility, microfilarial release by female worms and a MTT assay. The effect on the motility of adult worms was observed in a concentration- and time-dependent manner. The time required to stop motility of both sexes of adult worms was 6 h at 500 microg ml(-1), 24 h at 250 microg ml(-1), and 3 days for females and 4 days for males at 100 microg ml(-1). The movement of females ceased at 4 days at a concentration of 50 microg ml(-1) whereas the motility of males was only reduced. The loss of worm's viability was confirmed by the MTT assay and was similar to the motility results. These concentrations, including 10 microg ml(-1), prevented microfilarial release by females in a concentration- and time-dependent manner. Concentrations higher than 100 microg ml(-1) even induced mortality of the microfilariae. The present study suggested that the ethanol extract of Neurolaena lobata has potential macro- and micro-filaricidal activities. PMID- 15831110 TI - In vitro effects of four tropical plants on the activity and development of the parasitic nematode, Trichostrongylus colubriformis. AB - The in vitro effects of extracts of four tropical plants (Zanthoxylum zanthoxyloides, Newbouldia laevis, Morinda lucida and Carica papaya) on the egg, infective larvae and adult worms of Trichostrongylus colubriformis were screened for potential anthelmintic properties. Significant effects were observed with the four plants on T. colubriformis but they differed depending on the stage of the parasite. Extracts of each plant induced a dose-dependent inhibition of egg hatching. Using a larval inhibition migration test, the effects on the infective larvae were also detected with the four plant extracts. In contrast, for adult worms, the effects were statistically significant only for N. laevis and C. papaya. No significant activity was shown for M. lucida and Z. zanthoxyloides. These in vitro results suggest the presence of some anthelmintic properties associated with these four plants, which are traditionally used by small farmers in western Africa. These effects need to be studied under in vivo conditions. PMID- 15831111 TI - Gastrointestinal helminths of the lizard Chalcides ocellatus from Benghazi, Libya. AB - Chalcides ocellatus, a scincid lizard, sampled during October 1998 to December 1999 from Benina (farmland) and Al-Kueffia (a rocky area) in Benghazi, Libya, was found to harbour three intestinal nematodes, Pharyngodon mamillatus, Thelandros alatus and Parapharyngodon micipsae, and an intestinal cestode, Oochoristica tuberculata. Thelandros alatus, P. micipsae and O. tuberculata were recorded for the first time in C. ocellatus. Parapharyngodon micipsae was detected in C. ocellatus from Benina and O. tuberculata from lizards in Al-Kueffia. The majority (87.6%) of C. ocellatus were infected with helminth parasites and the levels of infection were higher in males than in females but this difference was not significant. Pharyngodon mamillatus recorded the highest number of nematode parasites in C. ocellatus, although there was no relationship between the number of nematode parasites recovered and host density. In lizards infected with the cestode O. tuberculata, males had shorter snout-vent lengths than females. From a total of 153 C. ocellatus, 120 (78.4%) showed single and 14 (9.2%) showed mixed parasitic infections. Mixed infections between different species of nematodes were not observed. PMID- 15831112 TI - Carbohydrate localization on Gyrodactylus salaris and G. derjavini and corresponding carbohydrate binding capacity of their hosts Salmo salar and S. trutta. AB - The congeners Gyrodactylus salaris and G. derjavini are specific ectoparasites of Atlantic salmon Salmo salar and brown trout S. trutta, respectively. To elucidate the involvement of lectin-carbohydrate interactions in this host specificity, carbohydrates on the tegument of the two species and the corresponding lectin activity of their hosts have been studied. Carbohydrate composition on the tegument differed significantly between the two gyrodactylids. Three of four commercially available peroxidase-labelled lectins with primary affinity towards D-mannoside, D-GalNAc and L-fucose bound more strongly to G. derjavini than to G. salaris. Lectins with an affinity towards D-mannoside and D-GalNAc bound significantly stronger to the cephalic lobes on G. derjavini compared to the tegument and sheaths of the hamuli. One brown trout strain and three different salmon strains were tested for lectin activity in skin and plasma. Two Baltic salmon strains and one strain from the Atlantic region were included. Brown trout differed significantly from the salmon strains when skin samples were tested for D-GalNAc activity. Lectins binding to other carbohydrates showed trends for similar host differences. The implications of carbohydrate-lectin interactions for host specificity in gyrodactylids are discussed. PMID- 15831113 TI - Life history constraints on the evolution of abbreviated life cycles in parasitic trematodes. AB - Abbreviations of the complex life cycle of trematodes, from three to two hosts, have occurred repeatedly and independently among trematode lineages. This is usually facultative and achieved via progenesis: following encystment in the second intermediate host, the metacercaria develops precociously into an egg producing adult, bypassing the need to reach a definitive host. Given that it provides relatively cheap insurance against a shortage of definitive hosts, it is not clear why facultative progenesis has only evolved in a few taxa. Here a comparative approach is used to test whether progenetic trematodes are characterized by larger body size and egg volumes, two traits that correlate with other key life history features, than other trematodes. These traits may constrain the evolution of progenesis, because precocious maturation might be impossible when the size difference between the metacercaria and a reproductive adult is too large. First, trematode species belonging to genera in which progenesis has been documented were found not to differ significantly from other trematode species. Second, using within-genus paired comparisons across 19 genera in which progenesis has been reported, progenetic species did not differ, with respect to body size or egg size, from their non-progenetic congeners. Third, using intraspecific paired comparisons in species where progenesis is facultative, no difference was observed in the sizes of eggs produced by worms in both the intermediate and definitive host, suggesting that opting for progenesis does not influence the size of a worm's eggs. Overall, the lack of obvious differences in body or egg size between trematodes with truncated life cycles and those with the normal three-host cycle indicates that basic life history characteristics are not acting as constraints on the evolution of progenesis; trematodes of all sizes can do it. Why facultative progenesis is not more widespread remains a mystery. PMID- 15831114 TI - Changes in survival characteristics of Diplostomum spathaceum cercariae emerged from cadmium-exposed Lymnaea stagnalis. AB - The effect of exposing Lymnaea stagnalis (Gastropoda: Pulmonata), infected with Diplostomum spathaceum (Trematoda: Diplostomatidae), to 100 microg l(-1) cadmium for 7 days on survival characteristics (survival, tail loss, decaudized cercarial life-span) of emerged cercariae was investigated. Exposure of L. stagnalis to cadmium resulted in significantly increased D. spathaceum cercarial survival and an inhibited tail loss compared to controls. The normal parallel relationship which exists over time between decreasing cercarial survival and increasing tail loss in controls was changed in cercariae from cadmium-exposed hosts with an increased proportion of cercarial deaths occurring without tail loss. The decaudized cercarial life-span over the survival period of the cercarial population did not significantly change. However comparisons between individuals decaudized during the initial 24 h time period with those which were decaudized during the final period of cercarial survival showed a significantly altered life span which did not occur in the control population. As a potential indicator of penetration 'fitness' comparisons were also undertaken between control and exposed cercariae decaudized during the initial 24 h time period, which revealed that the decaudized cercarial life-span from the exposed hosts was significantly different from controls. This may have important implications for the ability of cercariae to migrate through the tissues of their target host. The importance and relevance of these results to parasite transmission are discussed. PMID- 15831115 TI - Impact of health educational programmes on the prevalence of enterobiasis in schoolchildren in Thailand. AB - Enterobiasis is a worldwide prevalent disease particularly in low income areas. The budget needed for the prevention, treatment and eradication of the disease has thus far frustrated the limited budgets of global public health systems. A study was undertaken to determine if education in addition to medical treatment of enterobiasis could make a difference to the rates of infection. A total of 777 children (399 male and 378 female) from 11 elementary schools in five districts of Samut Prakan Province, Thailand were examined between December 2000 and March 2002. In five of the 11 schools studied, medical treatments were applied, followed by a programme of educating the children in the prevention of infection. Children in the remaining six schools received medical treatment only. The study showed a decrease in infections among children who received supplementary education. This decrease was significant in comparison to the decrease shown among children who received medical treatment only. The study therefore showed that educating high risk individuals played a key role in the prevention of enterobiasis. PMID- 15831116 TI - Ants as first intermediate hosts of Mesocestoides on San Miguel Island, USA. AB - This study tested the hypotheses that ants (Formicidae) function as a first intermediate host of Mesocestoides (Cestoda: Mesocestoididae) and that deer mice (Peromyscus maniculatus) develop metacestode infections after ingesting cysticercoid or procercoid-infected ants. Field studies were conducted at an island fox (Urocyon littoralis littoralis) breeding facility located on San Miguel Island, California Channel Islands National Park, USA, where > 40% of captive foxes were infected with adult Mesocestoides. Eight percent (8%) of deer mice at the fox pen site were infected with Mesocestoides metacestodes while none were infected at a distant site where foxes were absent (campground), thereby indicating the potential localized presence of a first intermediate host. To test whether ants from San Miguel Island contained Mesocestoides DNA, a polymerase chain reaction (PCR)-based diagnostic assay was developed using nested primers that could detect a single hexacanth larva within pooled samples of ten ants. Ants (Lasius niger and Tapinoma sessile) collected near the fox breeding facility were tested using the nested-PCR assay. Seven of 223 pooled samples of L. niger (3.1%) and 2 of 84 pooled samples of T. sessile (2.4%) tested positive for Mesocestoides DNA, while none of the ants were positive at the campground site. Positive samples were sequenced and found to match DNA sequences from Mesocestoides obtained from island fox and deer mice. Finally, to determine whether ants function as a first intermediate host for Mesocestoides, colony raised deer mice (n = 47) were fed L. niger (n = 3860) or T. sessile (n = 339) collected from the San Miguel Island fox breeding facility. No mouse became infected with Mesocestoides metacestodes after ingesting ants. While both L. niger and T. sessile from SMI were positive for Mesocestoides DNA, they were not infective to deer mice in the laboratory. PMID- 15831117 TI - Host population density as the major determinant of endoparasite species richness in floodplain fishes of the upper Parana River, Brazil. AB - A comparative analysis of parasite species richness was performed across 53 species of fish from the floodplain of the upper Parana River, Brazil. Values of catch per unit effort, CPUE (number of individuals of a given fish species captured per 1000 m(2) of net during 24 h) were used as a rough measure of population density for each fish species in order to test its influence on endoparasite species richness. The effects of several other host traits (body size, social behaviour, reproductive behaviour, spawning type, trophic category, feeding habits, relative position in the food web, preference for certain habitats and whether the fish species are native or exotic) on metazoan endoparasite species richness were also evaluated. The CPUE was the sole significant predictor of parasite species richness, whether controlling for the confounding influences of host phylogeny and sampling effort or not. The results suggest that in the floodplain of the upper Parana River (with homogeneous physical characteristics and occurrence of many flood pulses), population density of different host species might be the major determinant of their parasite species richness. PMID- 15831118 TI - Accumulation of the precious metals platinum, palladium and rhodium from automobile catalytic converters in Paratenuisentis ambiguus as compared with its fish host, Anguilla anguilla. AB - The platinum group metals (PGM) Pt, Pd and Rh are emitted into the environment mainly by catalytic exhaust gas converters of cars. As PGM accumulate in sediments of aquatic ecosystems, the study was focused on the uptake of the noble metals by European eels, Anguilla anguilla infected with the acanthocephalan Paratenuisentis ambiguus. Eels were exposed to ground catalytic converter material for six weeks. After exposure Pt and Pd were detected in the liver and kidney of the eels and in the parasites. Palladium was also found in fish muscle and intestine. No Rh uptake by the eel tissues and the parasites occurred. Paratenuisentis ambiguus contained the highest levels of both metals with 40 times higher Pt concentrations and four times higher Pd concentrations than the liver of its host. Due to its accumulation capacity for PGM, P. ambiguus can be applied as a sensitive accumulation indicator in field studies to assess the degree of environmental PGM contamination in aquatic ecosystems. PMID- 15831119 TI - Ligula intestinalis infection as a potential source of bias in the bioindication of endocrine disruption in the European chub Leuciscus cephalus. AB - European chub Leuciscus cephalus collected from five localities in the lowland and subalpine regions of Austria were analysed for oestrogenic effects of endocrine-disrupting chemicals and the presence of the plerocercoid of the tapeworm Ligula intestinalis. Of 1494 chub analysed, only seven (six males, one female) were found to be infected with single, but large plerocercoids up to 15 cm in length. Ligula-infected fish showed comparatively immature gonads, as demonstrated by the gonadosomatic index and gamete developmental stages. Plasma levels of the egg precursor protein vitellogenin also showed concentrations ranging below the detection limit. The present results indicate that chub infected with L. intestinalis and exposed to exogenous oestrogenic compounds can result in reduced gonadal maturation and produce false oestrogen-positive diagnoses in male fish. For plasma vitellogenin levels, L. intestinalis infections can result in false oestrogen-negative diagnoses in male and female fish. PMID- 15831120 TI - Food Standards Agency: nutrition. AB - The Food Standards Agency undertakes a range of activities with the aim of improving the dietary health of the population and determining how best to communicate key messages to achieve dietary change. Activities include obtaining sound evidence from research and surveys, and seeking advice from independent experts. Work is also undertaken to inform and motivate the population about diet and to identify ways of improving their diet. The effectiveness of the work is monitored in order to inform future policy decisions and interventions and to understand cost implications. PMID- 15831121 TI - Findings from the latest National Diet and Nutrition Survey. AB - The National Diet and Nutrition Survey (NDNS) of adults aged 19-64 years, carried out in 2000-1, is part of the NDNS programme, a series of cross-sectional surveys aiming to provide detailed quantitative information on the diet, nutritional status and related characteristics of the British population. The programme is split into four surveys of different population age-groups, conducted at approximately three-yearly intervals. In the survey of adults food consumption data were collected from 1724 respondents using a 7 d weighed-intake dietary record. Other components included: height, weight, waist and hip circumference and blood pressure measurements; a 24 h urine sample; a blood sample; a record of physical activity. Results have been published in four volumes covering food consumption, energy and macronutrient intakes, micronutrient intakes and nutritional status, including physical measurements and physical activity. The results have shown that, based on a comparison of nutrient intakes with the UK dietary reference values, adults in Britain are generally getting sufficient nutrients from their diets. However, younger adults (particularly women) and those in lower socio-economic groups are more likely to have low micronutrient intakes and lower levels of some nutritional status indices. The proportion of food energy derived from total fat has fallen since the last survey of this age group in 1986-7 and is close to the dietary reference value, while the proportion of energy derived from saturated fatty acids and non-milk extrinsic sugars exceeds the dietary reference values. The prevalence of overweight and obesity has increased since 1986-7 and physical activity levels are low. PMID- 15831122 TI - Are there functional consequences of a reduction in selenium intake in UK subjects? AB - Dietary Se levels in the UK have fallen over the last 20 years and recent surveys indicate that average Se intakes are 30-40 microg/d, which is well below the current UK reference nutrient intake for adult men (75 microg/d) or women (60 microg/d). Functional consequences of this decline have not been recognised, although epidemiological data suggest it may contribute to increased risk of infections and incidence of some cancers. Previous data have indicated that biochemical changes in Se-dependent proteins occur in otherwise healthy UK subjects given small Se supplements. The current studies have focused on the effect of small Se supplements on the immune response since there is evidence of specific interactions between Se intake and viral replication, and since the potential anti-cancer effects of Se may be mediated by non-antioxidant effects of Se such as changes in immune function. Data indicate that subjects given small Se supplements (50 or 100 microg Se/d) have changes in the activity of Se-dependent enzymes and evidence of improved immune function and clearance of an administered live attenuated virus in the form of poliovirus vaccine. Responses of individual subjects to Se supplements are variable, and current work is evaluating potential explanations for this variability, including genetic variability and pre-existing Se status. PMID- 15831123 TI - Iron nutrition in the UK: getting the balance right. AB - Fe homeostasis is considered in the context of the UK diet, using information on Fe intake and status from the National Diet and Nutrition Surveys. The importance of assessing Fe availability rather than total Fe intake is discussed. Dietary and host-related factors that determine Fe bioavailability (Fe utilised for Hb production) are reviewed using information from single-meal studies. When adaptive responses are taken into consideration, foods associated with higher Fe status include meat (haem-Fe and the 'meat factor') and fruits and fruit juice (vitamin C). Foods that may have a negative impact include dairy products (Ca), high-fibre foods (phytate) and tea and coffee (polyphenols), but the effects are more apparent in groups with marginal Fe deficiency, such as women of childbearing age. Analysis of dietary intake data on a meal-by-meal basis is needed to predict the influence of changing dietary patterns on Fe nutrition in the UK. Current information suggests that in the UK Fe deficiency is a greater problem than Fe overload. PMID- 15831124 TI - Folate bioavailability. AB - The achievement of optimal folate status to prevent neural-tube defects, and possibly other diseases, is hindered by the well-recognised incomplete bioavailability of the natural folates found in foods compared with the synthetic vitamin, folic acid. Folate bioavailability from different foods is considered to be dependent on a number of factors, including the food matrix, the intestinal deconjugation of polyglutamyl folates, the instability of certain labile folates during digestion and the presence of certain dietary constituents that may enhance folate stability during digestion. There is conflicting evidence as to whether the extent of conjugation of polyglutamyl folate (in the absence of specific inhibitors of deconjugation in certain foods) is a limiting factor in folate bioavailability. Estimates of the extent of lower bioavailability of food folates compared with folic acid (relative bioavailability) show great variation, ranging anywhere between 10 and 98%, depending on the methodological approach used. The lack of accurate data on folate bioavailability from natural food sources is of particular concern in those countries in which there is no mandatory folic acid fortification, and therefore a greater reliance on natural food folates as a means to optimise status. Apart from the incomplete bioavailability of food folates, the poor stability of folates in foods (particularly green vegetables) under typical conditions of cooking can substantially reduce the amount of vitamin ingested and thereby be an additional factor limiting the ability of food folates to enhance folate status. A recent workshop convened by the Food Standards Agency concluded that gaining a better understanding of folate bioavailability in representative human diets is a high priority for future research. PMID- 15831125 TI - Effecting dietary change. AB - A world epidemic of diet-related chronic disease is currently being faced. In the UK incidence of obesity alone has tripled in the last 20 years and this trend is predicted to continue. Consensus exists for the urgent need for a change in diet and other lifestyle factors and for the direction and targets for this change. The evidence for how this change can be achieved is less certain. It has been established that disease processes begin in childhood. Recent evidence indicates that dietary habits too are established in childhood but that these habits are amenable to change. While establishing a healthy lifestyle in childhood is paramount, interventions have the potential to promote positive change throughout the life course. Success in reversing current trends in diet-related disease will depend on commitment from legislators, health professionals, industry and individuals, and this collaboration must seek to address not only the food choices of the individual but also the environment that influences such choices. Recent public health policy development in England, if fully supported and implemented, is a positive move towards this goal. Evidence for effective strategies to promote dietary change at the individual level is emerging and three reviews of this evidence are discussed. In addition, three recent dietary intervention studies, in three different settings and with different methods and aims, are presented to illustrate methods of effecting dietary change. Further work is required on what factors influence the eating behaviour and physical activity of individuals. There is a need for further theory-based research on which to develop more effective strategies to enable individuals to adopt healthier lifestyles. PMID- 15831126 TI - Workshop on funding opportunities within the Food Standards Agency. AB - During this workshop, held as part of a joint Nutrition Society and Food Standards Agency (Agency) meeting on Micronutrient interactions and public health, several precepts for a successful funding application to the Agency were discussed. These precepts, many of which can be used as guiding principles for project proposals to other funding bodies, are summarised as follows: remember that the Agency only supports research that will help them formulate or change human food policy; read the research requirements document thoroughly and plan your project to answer the call; remember that the Agency issues contracts, not grants; your project will be just one project within a focused and coordinated programme; collaborative work is encouraged, but this type of approach is not a licence to double or treble your costs; write a one-page executive summary and attach it to the front of the form;the statistical basis for your experimental design and proposed statistical analysis of your results are important criteria in the evaluation of your proposal; your plans for dissemination and exploitation are very important;match your project duration against your research plan; abide by the Agency plan for quality assurance for the management of research; make full use of the programme adviser and the Agency policy contact and the 'feedback' stage to refine your scientific ideas in line with Agency policy. PMID- 15831127 TI - Iron, copper and fetal development. AB - Pregnancy is a period of rapid growth and cell differentiation for both the mother and fetus. Consequently, it is a period when both are vulnerable to changes in dietary supply, especially of those nutrients that are marginal under normal circumstances. In developed countries this vulnerability applies mainly to micronutrients. Even now, Fe deficiency is a common disorder, especially in pregnancy. Similarly, Cu intake in the UK population is rarely above adequate levels, which is a matter of some concern, both in terms of public health and possible clinical consequences. In early studies it was shown that lambs born to mothers on Cu-deficient pastures develop 'swayback,' with neurological and muscular symptoms that cannot be reversed by postnatal supplementation. More recently, rat studies have shown that responses such as the 'startle' response are lost in offspring of Cu-deficient mothers. Data have shown that prenatal Fe deficiency results in increased postnatal blood pressure, even though the offspring have normal dietary Fe levels from birth. These observations emphasise the importance of Fe and Cu in growth and development. In the present review the importance of these metals and the consequences, both short term and long term, of deficiency will be discussed and some possible mechanisms whereby these effects may be generated will be considered. PMID- 15831128 TI - The molecular basis of copper and iron interactions. AB - The intimate relationship between Fe and Cu in human nutrition has been recognised for many years. The best-characterised link is provided by caeruloplasmin, a multiCu-binding protein that acts as a serum ferrioxidase and is essential for the mobilisation of Fe from storage tissues. Decreased Cu status has been shown to reduce holo-caeruloplasmin production and impair ferrioxidase activity, leading, in a number of cases, to decreased tissue Fe release and the generation of anaemia that is responsive to dietary supplementation with Cu but not Fe. Dietary Fe absorption also requires the presence of a multiCu ferrioxidase. Hephaestin, a caeruloplasmin homologue, works in concert with the IREG1 transporter to permit Fe efflux from enterocytes for loading onto transferrin. The essential role of hephaestin in this process has been recognised from studies in the sex-linked anaemic (sla) mouse, in which Fe efflux is markedly impaired as a result of a mutation in the hephaestin gene that results in a truncated and non-functional version of the protein. There is emerging evidence that a number of other components of the intestinal Fe transport pathway are also Cu sensitive. Divalent metal transporter 1 (DMT1), the Fe transporter located at the apical membrane of enterocytes, is also a physiologically-relevant Cu transporter, suggesting that these two metals may compete with each other for uptake into the duodenal enterocytes. Furthermore, expression of both DMT1 and the basolateral Fe-efflux transporter IREG1 can be regulated by Cu, suggesting that the Fe-Cu relationship may be more complex than first thought. PMID- 15831129 TI - Folate, DNA stability and colo-rectal neoplasia. AB - Lower levels of dietary folate are associated with the development of epithelial cell tumours in man, particularly colo-rectal cancer. In the majority of epidemiological studies blood folate or reported folate intake have been shown to be inversely related to colo-rectal cancer risk. Folate, via its pivotal role in C1 metabolism, is crucial both for DNA synthesis and repair, and for DNA methylation. This function is compromised when vitamin B12 is low. Vitamin B12 deficiency has been shown to increase biomarkers of DNA damage in man but there is no evidence directly linking low vitamin B12 with cancer. Disturbingly, folate and vitamin B12 deficiencies are common in the general population, particularly in the underprivileged and the elderly. How folate and/or vitamin B12 deficiency influence carcinogenesis remains to be established, but it is currently believed that they may act to decrease DNA methylation, resulting in proto-oncogene activation, and/or to induce instability in the DNA molecule via a futile cycle of uracil misincorporation and removal. The relative importance of these two pathways may become clear by determining both DNA stability and cytosine methylation in individuals with different polymorphic variants of key folate metabolising enzymes. 5,10-Methylenetetrahydrofolate reductase converts 5,10 methylenetetrahydrofolate to 5-methyltetrahydrofolate and thereby controls whether folate is employed for DNA synthesis or DNA methylation. Colo-rectal cancer risk is decreased in subjects homozygous for a common variant (C677T) of the gene coding for this enzyme, suggesting that DNA synthesis and repair may be 'enhanced' in these individuals. Evidence from animal and human studies is presented here in support of folate acting to maintain genomic stability through both these mechanisms. PMID- 15831130 TI - Dietary antioxidants and environmental stress. AB - Air is one of our most important natural resources; however, it is also in the front line for receiving environmental pollution. Air quality decreased markedly following the industrial revolution, but it was not until the great London Smog in 1952 that air quality made it onto the political agenda. The introduction of the Clean Air Act in 1956 led to dramatic decreases in black smoke and SO2 concentrations over the next two decades, as domestic and industrial coal-burning activities ceased. However, as these improvements progressed, a new threat to public health was being released into the air in ever-increasing quantities. Rapid motorisation of society from the 1960s onwards has led to the increased release of atmospheric pollutants such as tiny particles (particulate matter of <10 microm in aerodynamic diameter) and oxides of N, and the generation of the secondary pollutant O3. These primary and secondary traffic-related pollutants have all proved to be major risks factors to public health. Recently, oxidative stress has been identified as a unifying feature underlying the toxic actions of these pollutants. Fortunately, the surface of the lung is covered with a thin layer of fluid containing a range of antioxidants that appear to provide the first line of defence against oxidant pollutants. As diet is the only source of antioxidant micronutrients, a plausible link now exists between the sensitivity to air pollution and the quality of the food eaten. However, many questions remain unanswered in relation to inter-individual sensitivity to ambient air pollution, and extent to which this sensitivity is modified by airway antioxidant defences. PMID- 15831131 TI - Micronutrients and cancer. AB - Adverse dietary factors account for approximately 30% of all cancers. Overconsumption of energy is undoubtedly one of the major risk factors, but dietary composition is also very important. In particular, a low consumption of fruits and vegetables appears to double the risk of carcinomas of the lung and alimentary tract. Epidemiological studies suggest that high plasma levels of Se, carotenoids and ascorbic acid are protective against cancer. However, intervention studies with antioxidant nutrients have given mixed results, and it has not been established that the benefits of a high intake of fruits and vegetables are invariably related to the prevention of oxidative DNA damage. Folic acid supplementation appears to protect against colo-rectal neoplasia, probably by preventing mutations associated with the repair of uracil mis incorporation. However, there are indications from animal studies that exposure to high levels of folic acid at certain stages of development may lead to epigenetic effects that are, as yet, poorly understood. There seems little doubt that micronutrients contribute to the protective effects of plant foods against cancers of the lung and alimentary tract, but it has not been established that these benefits can be achieved using supplements. PMID- 15831132 TI - B-vitamins, homocysteine metabolism and CVD. AB - The present review focuses on the B-vitamins, i.e. folate, vitamin B12, vitamin B6 and riboflavin, that are involved in homocysteine metabolism. Homocysteine is a S-containing amino acid and its plasma concentrations can be raised by various constitutive, genetic and lifestyle factors, by inadequate nutrient status and as a result of systemic disease and various drugs. Hyperhomocysteinaemia is a modest independent predictor of CVD and stroke, but causality and the precise pathophysiological mechanism(s) of homocysteine action remain unproven. The predominant nutritional cause of raised plasma homocysteine in most healthy populations is folate insufficiency. Vitamin B12 and, to a lesser extent, vitamin B6 are also effective at lowering plasma homocysteine, especially after homocysteine lowering by folic acid in those individuals presenting with raised plasma homocysteine. However, riboflavin supplementation appears to be effective at lowering plasma homocysteine only in those individuals homozygous for the T allele of the C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene. This gene codes for the MTHFR enzyme that produces methyltetrahydrofolate, which, in turn, is a substrate for the remethylation of homocysteine by the vitamin B12-dependent enzyme methionine synthase. Individuals with the MTHFR 677TT genotype are genetically predisposed to elevated plasma homocysteine, and in most populations have a markedly higher risk of CVD. PMID- 15831133 TI - Public health aspects of food fortification: a question of balance. AB - Micronutrient malnutrition is widespread throughout the world, with important health and economic consequences. Tools to address this situation include food fortification, supplementation and dietary diversification, each having different and complementary roles. Fortification (mandatory and voluntary) has been practised over several decades in Western countries as well as in developing countries. Iodised salt was introduced in the USA in 1924 to reduce severe I deficiency. In 1938 voluntary enrichment of flours and breads with niacin and Fe was initiated to reduce the incidence of pellagra and Fe-deficiency anaemia respectively. Micronutrient intakes in European countries appear to be generally adequate for most nutrients. However, a number of population subgroups are at higher risk of suboptimal intakes (below the lower reference nutrient intake) for some micronutrients, e.g. folate, Fe, Zn and Ca in children, adolescents and young women. Dietary surveys indicate that fortified foods play a role in mitigating such risks for several important nutrients. The number of foods suited to fortification are considerably limited by several factors, including technological properties (notably moisture, pH and O2 permeability), leading to unacceptable taste and appearance, as well as cost and consumer expectations. In countries in which voluntary fortification is widely practised micronutrient intakes are considerably below tolerable upper intake levels. Concerns about safety are addressed in relation to the potentially increased level or proportion of fortified foods (e.g. following potential EU legislation), for nutrients with relatively low tolerable upper intake levels and where the potential benefit and risks are in different subpopulations (e.g. folic acid). Recent models for assessing these issues are discussed. PMID- 15831134 TI - The evolution of the nutritional management of diabetes. AB - Diet and lifestyle advice for individuals with diabetes has changed dramatically. The changes in nutritional recommendations have largely been in response to advances in the knowledge of the biochemical and physiological mechanisms of impaired glucose metabolism and the micro- and macrovascular complications of diabetes. The most recent guidelines for the nutritional management of diabetes were set out by Diabetes UK in 2003. This consensus-based advice paper builds on the previous evidence-based review papers of the European Association for the Study of Diabetes in 2000 and the American Diabetes Association in 2002. The changes from previous recommendations include a more liberal use of sucrose, in line with healthy eating recommendations for the general population, and allow for greater flexibility in energy derived from carbohydrate and monounsaturated fat. In addition, monounsaturated fats are promoted as the fat of choice and active promotion of carbohydrate foods with a low glycaemic index is encouraged. These guidelines emphasise the practical application of nutritional management of diabetes and the need to provide education and support in a structured way that will facilitate change in diet and lifestyle behaviour. Structured educational programmes have been shown to be effective in reducing the progression to diabetes and also in slowing the onset and progression of the complications of diabetes. These programmes require ongoing intensive input to maintain behavioural change in diet and lifestyle. Considerable energy and resources are required to set up and maintain these educational programmes, but the cost per individual is small compared with the costs of treating the complications of diabetes. PMID- 15831135 TI - Antimicrobial properties of plant secondary metabolites. AB - Increasing awareness of hazards associated with the use of antibiotic and chemical feed additives has accelerated investigations into plants and their extracts as feed additives. The present review mainly discusses two classes of plant secondary compounds in this context, i.e. essential oils and saponins. The broader potential of plants and their extracts is illustrated by the progress of an EC Framework 5 project, 'Rumen-up'. Dietary inclusion of a commercial blend of essential oils causes markedly decreased NH3 production from amino acids in rumen fluid taken from sheep and cattle. This effect is mediated partly by the effects on hyper-NH3-producing bacteria and the protein- and starch-fermenting rumen bacterium, Ruminobacter amylophilus. Saponin-containing plants and their extracts suppress the bacteriolytic activity of rumen ciliate protozoa and thereby enhance total microbial protein flow from the rumen. The effects of some saponins are transient, because saponins are hydrolysed by bacteria to their corresponding sapogenin aglycones, which are much less toxic to protozoa. Saponins also have selective antibacterial effects that may prove useful in, for example, controlling starch digestion. The 'Rumen-up' project began with a targetted collection of European plants and their extracts, which partners have tested for their effects on rumen proteolysis, protozoa, methanogenesis and lactate production. A success rate of about 5% in terms of positive hits illustrates that plant secondary compounds, of which essential oils and saponins comprise a small proportion, have great potential as 'natural' manipulators of rumen fermentation to benefit the farmer and the environment in the future. PMID- 15831136 TI - Plant secondary metabolites: antiparasitic effects and their role in ruminant production systems. AB - The purported antiparasitic properties of plant secondary metabolites (PSM) have been the cause of controversy amongst the scientific community. Despite long standing knowledge of the prophylactic and therapeutic properties of PSM-rich extracts, which comes mainly from ethnoveterinary sources, the scientific evidence of the antiparasitic effects of PSM is inconsistent. In the first part of the present paper the causes of this controversy are addressed, and the evidence available on the antiparasitic effects of PSM is critically examined. The focus is on examples of the antiparasitic activity of PSM against helminth nematodes. The conclusion is that PSM can have antiparasitic properties, which depend on their structure, level of ingestion and availability within the gastrointestinal tract of the animal. The second part is an appraisal of the potential role of PSM for parasite control in ruminant production systems. Despite their antiparasitic properties, PSM consumption does not necessarily have positive consequences in parasitised herbivores, as excessive consumption of PSM can adversely affect herbivore fitness and survival, through their anti nutritional properties. For this reason it is suggested that the antiparasitic properties of PSM should be assessed at the same time as their anti-nutritional effects. The same measure, e.g. the performance of parasitised hosts, should be used when assessing these properties. The assessment of the costs and benefits suggests that parasitised herbivores can benefit from the long-term consumption of PSM only if the antiparasitic benefits outweigh the anti-nutritional costs of PSM. In addition, it is proposed that parasitised animals might be able to benefit from PSM consumption even if their performance is impaired, as long as the latter is a short-term compromise that leads to long-term benefits. PMID- 15831137 TI - The principles and application of qualitative research. AB - The present paper provides an overview of the methodological principles that underpin qualitative research and how these principles differ from those of quantitative research. It is intended to set the scene for the following papers that outline two specific approaches to the analysis of qualitative data. Within the tradition of qualitative research there are many different theoretical perspectives, of which these approaches are only two examples, but they need to be set within this broader tradition in order to highlight their specific features. Qualitative and quantitative research differ from each other in far more than their methods and data. They are each based on very different premises about both the nature of the world and the nature of our knowledge of it and how this information is generated. These approaches have implications for all aspects of research strategy, including the assessment of the quality of research findings and their wider utility or application. In relation to the latter, lack of detail in the reporting of qualitative research and small sample sizes has tended to create the impression that the findings of qualitative research have little application outside the particular research setting. While there is need for more rigor in reporting, it needs to be recognized that qualitative research can offer insights and understandings with wider relevance, although these outcomes are of a different type from those provided by quantitative research. PMID- 15831138 TI - Using interpretative phenomenological analysis for public health nutrition and dietetic research: a practical guide. AB - The present paper introduces interpretative phenomenological analysis (IPA) as a framework for analysing qualitative research data collected for public health nutrition and dietetic research studies. The theoretical roots of IPA have been set out briefly in order to help researchers decide whether the approach is relevant for them and their particular research questions. IPA can be used to analyse data from one-to-one interviews in order to develop 'thick descriptions' that may help illuminate human experience. IPA can also be used to develop theories and/or models, which could help inform policy. The main body of the paper describes the analytical techniques and procedures used to achieve both outcomes in a practical way, using examples from the author's own research. In the following paper Krueger's methods for analysing focus group data are set out for comparison. It is hoped that these papers will empower researchers with little experience of qualitative research to develop confidence with qualitative data analysis. In addition, it is hoped that the material will stimulate debate amongst more experienced qualitative researchers from a public health or dietetic background. PMID- 15831139 TI - Focus-group interview and data analysis. AB - In recent years focus-group interviews, as a means of qualitative data collection, have gained popularity amongst professionals within the health and social care arena. Despite this popularity, analysing qualitative data, particularly focus-group interviews, poses a challenge to most practitioner researchers. The present paper responds to the needs expressed by public health nutritionists, community dietitians and health development specialists following two training sessions organised collaboratively by the Health Development Agency, the Nutrition Society and the British Dietetic Association in 2003. The focus of the present paper is on the concepts and application of framework analysis, especially the use of Krueger's framework. It provides some practical steps for the analysis of individual data, as well as focus-group data using examples from the author's own research, in such a way as to assist the newcomer to qualitative research to engage with the methodology. Thus, it complements the papers by Draper (2004) and Fade (2004) that discuss in detail the complementary role of qualitative data in researching human behaviours, feelings and attitudes. Draper (2004) has provided theoretical and philosophical bases for qualitative data analysis. Fade (2004) has described interpretative phenomenology analysis as a method of analysing individual interview data. The present paper, using framework analysis concentrating on focus-group interviews, provides another approach to qualitative data analysis. PMID- 15831140 TI - Ayurveda and Yoga in Cardiovascular Diseases. AB - Ayurveda is derived from 2 Sanskrit words, namely, "Ayus" and "Veda," meaning life and knowledge, respectively. It literally means science of life. Ayurveda, of which yoga is an integral part, is widely practiced in India and is gaining acceptance in many countries around the world. It is a comprehensive and a holistic system, the focus of which is on the body, mind, and consciousness. The Ayurvedic treatment consists of the use herbal preparations, diet, yoga, meditation, and other practices. Based on the review of available studies, the evidence is not convincing that any Ayurvedic herbal treatment is effective in the treatment of heart disease or hypertension. However, the use of certain spices and herbs such as garlic and turmeric in an overall healthy diet is appropriate. Many herbs used by Ayurvedic practitioners show promise and could be appropriate for larger randomized trials. Yoga, an integral part of Ayurveda, has been shown to be useful to patients with heart disease and hypertension. Yoga reduces anxiety, promotes well-being, and improves quality of life. Its safety profile is excellent. Its use as a complementary therapeutic regimen under medical supervision is appropriate and could be worth considering. PMID- 15831141 TI - A man with syncope and ST segment elevation. PMID- 15831142 TI - Effusive-constrictive calcific pericarditis associated with Streptococcus salivarius. Case report and review of the literature. AB - We report the case of a 40-year-old patient presenting with a 6-month history of dyspnea and edema, with significant worsening of his clinical manifestations for the 2 weeks before admission to our department. During this 14-day preadmission period, continuous positive airway pressure (CPAP) was prescribed elsewhere for management of a working diagnosis of obstructive sleep apnea. The patient presented to us hemodynamically compromised. Management of the patient included emergency cardiac surgery for tamponade caused by effusive-constrictive, calcific pericarditis in addition to antimicrobial treatment as a result of the growth of Streptococcus salivarius from the pericardial fluid. This is the first report in the literature of association of this microorganism with pericarditis. The use of CPAP made the patient's symptoms worse as a result of an increase of the intrathoracic pressure, which was a pathophysiological mechanism, added to the interference of the localized pericardial effusion and the effect of the pericardial constriction. In an era of rapidly increasing use of CPAP systems, clinicians should be aware of their possible detrimental effects on patients with some types of cardiopulmonary diseases. PMID- 15831143 TI - Interactions of high salt intake and the response of the cardiovascular system to aldosterone. AB - High salt intake contributes to the risk of hypertension, and this effect is in part mediated by the physiologic action of aldosterone on renal mineralocorticoid receptors. However, the actions of aldosterone are not restricted to the kidneys, because aldosterone can bind to mineralocorticoid receptors in the heart, vasculature, and brain to produce structural and functional changes that lead to target organ damage. Experimental and clinical studies show that, in the setting of high salt intake, blocking aldosterone at the mineralocorticoid receptor reduces progression to target organ damage and preserves vascular function. In many cases, these benefits are independent of changes in blood pressure. Angiotensin-converting enzyme inhibitors and angiotensin receptor blockers have short-term effects on reducing aldosterone levels, but frequently aldosterone levels return to pretreatment levels during long-term therapy. Aldosterone blockade may be more completely achieved with mineralocorticoid receptor antagonists. Spironolactone has been shown to have substantial and significant benefits in experimental and clinical studies of cardiac dysfunction. Eplerenone is a selective aldosterone blocker with a greater binding affinity for mineralocorticoid receptors than for androgen and progesterone receptors. Eplerenone has similarly demonstrated significant benefits in experimental animals and in patients with left ventricular dysfunction after myocardial infarction. Thus, aldosterone blockade with mineralocorticoid receptor antagonists offers target organ protection and may blunt some of the adverse effects of chronic high salt intake. PMID- 15831144 TI - A rare case of Actinomyces israelii presenting as pericarditis in a 75-year-old man. AB - Actinomyces israelii is a gram-positive bacillus that is rarely associated with infections in the general population. A. israelii belongs to the normal flora of the body and it rarely becomes pathogenic. Cardiac involvement is rare and in most cases involves the pericardium. Fewer than 20 cases of pericardial actinomycosis have been reported in the literature since 1950. We report the case of a 75-year-old man with a history of coronary artery disease with recent myocardial infarction and stent placement, atrial fibrillation, and recent colonic perforation with subsequent colectomy/colostomy who presented to our hospital with a 2-week history of left-sided chest pain. Workup revealed the presence of a pericardial effusion and pericarditis. Pericardial fluid analysis demonstrated A. israelii. An examination and discussion of the literature is performed regarding this rare manifestation of human actinomycosis. PMID- 15831145 TI - Endovascular repair of symptomatic coronary-subclavian steal syndrome due to stenosis of the proximal left subclavian artery. AB - We report a patient with left subclavian artery stenosis in whom the internal thoracic artery (ITA) had been used as a coronary bypass. She presented with symptomatic myocardial and brain ischemia resulting from coronary-subclavian steal syndrome and was successfully treated with angioplasty and stenting. PMID- 15831146 TI - Device therapy in the management of congestive heart failure. AB - Despite significant advancements in the treatment of heart failure over the past 2 decades, this patient population is still subject to considerably high morbidity and mortality rates. In recent years, the field of device therapy as adjunctive treatment to the medical management of congestive heart failure has grown in the wake of the large number of randomized trials that have demonstrated the safety and efficacy of these devices. The implantable defibrillator currently represents the standard of care in certain segments of the heart failure population, even in those without a prior arrhythmic event. Biventricular pacing systems appear to have a role in heart failure patients with prolongation of their QRS duration in improving ventricular performance and symptoms, if not mortality. Last, the shortage of organs available for orthotopic transplant has heightened interest in using ventricular-assist devices as destination therapy, and although there is evidence for the feasibility for this approach at the current time, there is a next generation of devices that appear even more promising. PMID- 15831147 TI - Intracoronary brachytherapy for treatment of in-stent restenosis. AB - Randomized, double-blind, placebo-controlled trials have demonstrated that intracoronary brachytherapy is more efficacious than placebo in reducing death, myocardial infarction, and target vessel revascularization at long-term follow up of patients with in-stent restenosis. Intracoronary brachytherapy is efficacious in treating totally occluded in-stent restenotic lesions, in treating de novo and in-stent restenotic lesions in saphenous vein grafts, in treating diffuse in stent restenosis, in treating native coronary ostial in-stent restenotic lesions, in treating patients with diabetes with in-stent restenosis, in treating patients at high-risk for recurrence of restenosis, in treating elderly patients, and in treating patients who failed intracoronary radiation. Beta and gamma intracoronary brachytherapy are equally effective in treating in-stent restenosis. Long-term aspirin and clopidogrel should be administered for at least 1 year to reduce late vessel thrombosis. Inadequate radiation may cause edge stenosis. PMID- 15831148 TI - The cardiac ryanodine receptor (RyR2) and its role in heart disease. AB - The cardiac ryanodine receptor has become a subject of increasing interest as its role in the etiology of cardiac disease is becoming more apparent. In this article, we review the current knowledge of the structure and function of the cardiac ryanodine receptor and its implications in cardiac pathophysiology. Cardiac ryanodine receptors function by regulating calcium release from the sarcoplasmic reticulum in cardiomyocytes, thereby playing an integral role in excitation-contraction coupling. In heart failure, the myocardium remains in a chronic hyperadrenergic state. This leads to protein kinase A hyperphosphorylation of ryanodine receptors within cardiomyocytes, ultimately leading to calcium leakage from the sarcoplasmic reticulum into the cytosol and thus impairing excitation-contraction coupling. These mechanisms could partially explain the pathophysiology underlying the reduced cardiac output seen in heart failure. Beta-adrenergic blockade appears to correct the abnormality and reestablishes normal ryanodine receptor function. These calcium leaks can also generate delayed afterdepolarizations, which can lead to fatal arrhythmias. Two genetic diseases have been linked to mutations in the cardiac ryanodine receptor: arrhythmogenic right ventricular dysplasia type 2 and catecholaminergic polymorphic ventricular tachycardia or familial polymorphic ventricular tachycardia. As our understanding of this receptor and its modulators deepens, the possibility of clinical application draws near. PMID- 15831150 TI - Mitral valve endocarditis resulting from coagulase-negative Staphylococcus after stent implantation in a saphenous vein graft. AB - Valvular endocarditis after percutaneous coronary intervention is unusual. We report a new case of mitral valve endocarditis after stent implantation to a saphenous vein graft. PMID- 15831149 TI - Appreciation of precordial cardiac murmur on examination relative to knowledge of valvular heart disease. AB - Skills associated with physical examination have continued to decline as practitioners have become increasingly reliant on advanced imaging modalities. Our study sought to determine if documentation of valvular heart disease on echocardiography resulted in an increased appreciation and documentation of precordial murmur. PMID- 15831151 TI - Persistence with a persistent left and absent right superior vena cava. PMID- 15831152 TI - The fall-out from the Bristol Inquiry. PMID- 15831153 TI - Haemodynamic changes during modified ultrafiltration immediately following the first stage of the Norwood reconstruction. AB - BACKGROUND: Modified ultrafiltration has been shown to reverse haemodilution and improve ventricular function following cardiopulmonary bypass. There has been concern, however, about the safety and efficacy of modified ultrafiltration after the first stage of Norwood reconstruction for palliation of neonates with hypoplasia of the left heart and its variants. METHODS: We reviewed the intraoperative course of all patients undergoing the first stage of Norwood reconstruction between September 1, 2000, and August 31, 2002. RESULTS: The first stage of reconstruction was performed in 99 neonates, 78 with classical hypoplasia of the left heart, and 21 with variants. Mean weight at surgery was 3.1 plus or minus 0.7 kilograms. Genetic syndromes, weight less than or equal to 2.5 kilograms, and/or major additional cardiac or non-cardiac anomalies, were present in 44 patients. We deemed these patients to constitute the group at high risk. A modified Blalock-Taussig shunt was utilized in 95 patients, and a conduit from the right ventricle to the pulmonary arteries in 4. Deep hypothermic circulatory arrest was used in all patients for a mean period of 45 minutes, plus or minus 15 minutes. Total support time on cardiopulmonary bypass plus deep hypothermic circulatory arrest was 100 minutes plus or minus 26 minutes. Modified ultrafiltration was performed in all patients. The mean duration of modified ultrafiltration was 10 plus or minus 2 minutes, and the total volume of filtrate removed was 104 plus or minus 29 millilitres per kilogram. There were no complications from modified ultrafiltration, and no patient required discontinuation of modified ultrafiltration for haemodynamic instability. During modified ultrafiltration, the haematocrit increased from 31 percent plus or minus 4 to 46 percent plus or minus 6. Heart rate decreased from 170 plus or minus 17 beats per minute to 158 plus or minus 16 beats per minute. Systolic blood pressure increased from 57 plus or minus 12 to 63 plus or minus 13 millimetres of mercury, and diastolic blood pressure from 30 plus or minus 8 to 35 plus or minus 7 millimetres of mercury. All these values are significant at a p value of less than 0.0001. Hospital morality in the patients at low risk was 3 of 55 (5.5 percent), but was 12 of 44 (27.3 percent) in the patients deemed to be at high risk. CONCLUSIONS: Modified ultrafiltration is safe procedure following the first stage of Norwood reconstruction, with improvement in all haemodynamic parameters measured. Modified ultrafiltration is an additional incremental strategy, which may contribute to the overall improvement in outcome following surgical palliation of patients with hypoplasia of the left heart or its variants. PMID- 15831154 TI - Is heart rate variability an objective parameter with which to manage treatment of infants with heart failure due to left-to-right shunting? AB - Treatment in heart failure could be guided by additional non-clinical measures, such as neurohumoral levels. Variability in heart rate is known to reflect neurohumoral stimulation. With this in mind, we sought to assess retrospectively the variability in heart rate to guide the treatment of infants in heart failure. We analysed retrospectively the data from 20 infants with a significant left-to right shunt. All were unsuitable for cardiac surgery or interventional therapy at the time the treatment had commenced. None of the infants improved while receiving diuretics, spironolactone, and digoxin alone, but improved after the addition of propanolol or metoprolol. None of the infants had problems during or after the subsequent operation. Parasympathetic activity reflected by parameters of variability in heart rate, such as the square root of adjacent RR-intervals, and the amount of adjacent RR-intervals greater than 50 milliseconds, improved in nearly all infants during beta blockade. On the other hand, parameters of variability in heart rate reflecting sympathetic activity did not change. Parasympathetic activity reflected the clinical state of nearly all the infants. These parameters, therefore, seem to be a good non-clinical parameter, showing the optimal treatment for heart failure in an ambulatory setting. PMID- 15831155 TI - Amiodarone for post-operative junctional ectopic tachycardia. AB - BACKGROUND: Post-operative junctional ectopic tachycardia is a transient, but potentially life threatening, rapid automatic tachyarrhythmia that requires urgent and adequate treatment. In our study, we review retrospectively the use and efficacy of amiodarone for this arrhythmia over an 8-year period in our institution. METHODS AND PATIENTS: Retrospective review revealed 15 patients who were administered amiodarone for post-operative junctional ectopic tachycardia during the period. The median age was 2.6 months, with a range from 8 days to 8.1 months. The median weight was 4.6 kilograms, with a range from 2.6 to 8.2 kilograms. RESULTS: The median heart rate at diagnosis of the tachycardia was 192 beats per minute, and the range was 182 to 229 beats per minute. The biochemistry was essentially normal. The median length of time until the tachycardia was controlled was 4.5 hours, and the range was from 1 to 19.5 hours, with 13 of the 15 patients controlled within 12 hours. The median dose of amiodarone received by this time was 5.9 milligrams per kilogram, with a range from 1.0 to 25.0 milligrams per kilogram. Hypotension or bradycardia within 4 hours of commencing amiodarone were noted in 2 patients. CONCLUSION: Experience in our institution, and a review of the literature, suggests that the most rapid control of post operative junctional ectopic tachycardia will be obtained by a bolus of amiodarone followed by an intravenous infusion. Intravenous amiodarone is generally safe, with few side effects. Reported life threatening arrhythmias, however, suggest that intravenous amiodarone should be restricted to a setting where invasive monitoring and external cardiac pacing are available. PMID- 15831156 TI - Analysis of prothrombotic mutations and polymorphisms in children who developed thrombosis in the perioperative period of congenital cardiac surgery. AB - In this study, we investigated some of the prothrombothic mutations and polymorphisms in 15 children with congenital cardiac malformations who developed severe thrombosis in the perioperative period following surgical repair. The mutations and polymorphisms included in the study were Factor V Leiden, prothrombin G20210A, methylentetrahydrofolate reductase C677T, endothelial nitric oxide synthase intron 4 VNTR, alpha-fibrinogen Thr312Ala, Factor XIII Val34Leu, and insertion or deletion of angiotensin 1 converting enzyme. Compared to the healthy Turkish subjects, our patients had a similar rate of mutation of Factor V Leiden, Factor XIII Val34Leu, and endothelial nitric oxide synthase a/b polymorphisms, but higher frequency of the prothrombotic angiotensin 1 converting enzyme deletion/deletion genotype, and lower frequency of the antithrombotic alpha fibrinogen Thr/Thr genotype. None of the patients exhibited mutations involving prothrombin G20210A or methylentetrahydrofolate reductase C677T. The results of our study suggest that, in addition to prothrombotic mutations such as Factor V Leiden, single-nucleotide polymorphisms should be considered in all children with congenital cardiac malformations who develop thrombosis. Malformations of the heart are the most common of all serious lesions that are present at birth, with an incidence of 4 to 8 cases per 1,000 live births. If needed, corrective surgery is usually the optimal treatment for these anomalies, but perioperative morbidity and mortality still remain high due to several factors. Arterial or venous thrombosis, or both varieties of thrombosis, is among these factors. Prior to surgery, the most frequent time at which these children develop thrombosis is during cardiac catheterization. Postoperative thrombosis in this group of patients is a more complex disorder, which can affect both small and large vessels, and is associated with a high morbidity and mortality. Recent studies indicate that both point mutations and single-nucleotide polymorphisms of genes that encode proteins involved in the coagulative and anticoagulative cascades are important risk factors for development of thrombosis. Patients with these risk factors are most likely to develop thrombosis when triggering elements, such as placement of catheters, prolonged immobilization, or surgery, are also present. In this study, we investigated some of the above-mentioned mutations and polymorphisms in children who developed thrombosis in the perioperative period after correction of congenital cardiac malformations. PMID- 15831157 TI - Evidence of endothelial dysfunction in patients with functionally univentricular physiology before completion of the Fontan operation. AB - INTRODUCTION: Postoperative thrombosis after a cavopulmonary connection has been widely described. Abnormalities in coagulation seem to occur early in the course of patients with functionally univentricular physiology, and may precede surgery. Endothelial abnormalities due to chronic hypoxia, and hyperviscosity, may contribute to this scene. The purpose of our study was to investigate if patients with a superior cavopulmonary connection have altered levels of endothelial and coagulative markers in the plasma. METHODS: We compared findings in 10 patients, aged from 4 to 19 years, with 6 age-matched normal controls. We measured levels of von Willebrand factor antigen, thrombomodulin, tissue-type plasminogen activator, plasminogen activator inhibitor-1 and d-dimer in the plasma using enzyme-linked immunosorbent assay. RESULTS: We found increased levels of von Willebrand factor antigen (p = 0.01), tissue-type plasminogen activator (p = 0.01), and decreased levels of thrombomodulin (p = 0.008) in the patients when compared to controls, while levels of plasminogen activator inhibitor-1 were not different. Values of d-dimer were within the reference range. Levels of tissue type plasminogen activator had a positive correlation with von Willebrand factor antigen (r = 0.66, p = 0.008). CONCLUSIONS: Altered levels of endothelial markers in the plasma, in the presence of normal levels of d-dimer, suggest that endothelial dysfunction may precede the occurrence of intravascular coagulation and thrombosis in patients with functionally univentricular physiology. These observations may have therapeutical implications. PMID- 15831158 TI - A novel technique for creation of a systemic-to-pulmonary arterial shunt. AB - A systemic-to-pulmonary arterial shunt is still widely used for palliation of some neonates with cyanotic congenital cardiac lesions. This procedure, however, is well known to be associated with some degree of morbidity and mortality. To reduce the incidence of iatrogenic pulmonary arterial deformities, we have devised a new and simple technique to create the shunt using a partial sternotomy, and have used our technique in 10 neonates with cyanotic cardiac malformations. All but one of our patients survived, and an early reoperation was needed in only one further patient for revision of the shunt. Successful bidirectional Glenn procedures were performed in five of the nine surviving patients within 18 months of the initial procedure. We believe that our technique provides superior palliation by permitting equal enlargement of the right and left pulmonary arteries, and thus facilitates subsequent completion of the Fontan circulation. PMID- 15831159 TI - The implications for fetal outcome of an abnormal arrangement of the abdominal vessels. AB - In a prospective audit of consecutive referrals for fetal echocardiography between 1997 and 2003, we documented all instances of an abnormal arrangement of the abdominal vessels. We then established the structure of the heart in these fetuses, noting any extra-cardiac associations, and the eventual outcomes. We found a cardiac abnormality in 572 of the 2,136 fetuses examined during this period (27 percent), with 16 (0.8 percent) having an abnormal arrangement of the abdominal great vessels. Mirror-imaged arrangement was found in 3, while the arrangement suggested right isomerism in 6, and left isomerism in 7. Of these 16 fetuses, 14 had cardiac malformations. Isomerism of the right atrial appendages was found in 7 fetuses, all with either a right-sided stomach or cardiac apex, and 6 with a common atrioventricular junction guarded by a common valve. Only 1 of these fetuses survived. Of the 3 fetuses with mirror-imaged abdominal great vessels, 2 also had mirror-imaged atrial arrangement, while the 3rd had isomeric right appendages. Only one of 7 fetuses with an abdominal great vein posterior to the aorta had bilateral left atrial appendages. The remaining 6 had usual atrial arrangement, with normal pulmonary venous connections in all but one, who had infra-diaphragmatic totally anomalous pulmonary venous connection. Of these fetuses, 2 had coarctation of the aorta, and 2 others had complex cardiac malformations resulting in neonatal death. One died in childhood from biliary atresia, and three are alive. Abnormal arrangements of the abdominal great vessels, therefore, were found in 0.8 percent of our total fetuses, and in 2.4 percent of those with cardiac malformations. Those with an abnormal arrangement of the abdominal vessels combined with an abnormal arrangement of the atrial appendages, however, made up only 0.5 percent and 1.6 percent, respectively. While screening for an abnormal arrangement of the abdominal vessels overestimated the incidence of left, but not right, isomerism of the atrial appendages, it did predict the presence of important extra-cardiac malformations that required urgent recognition and management after birth. PMID- 15831160 TI - New outpatient referrals to a tertiary paediatric cardiac centre: evidence of increasing workload and evolving patterns of referral. AB - OBJECTIVES: To assess the volume and range of diagnosis in new patients referred to paediatric cardiac outpatient clinics. METHODS: Data was collected prospectively, using a proforma completed at all outpatient clinics over a period of three months. RESULTS: There were 526 new referrals, representing an increase of almost one-fifth compared to 5 years ago. Of the referrals, 78 percent came from hospital doctors, and 22 percent from general practitioners, with 221 of those referred being infants. A heart murmur was the most common reason for referral, representing almost two-thirds of cases. In 372 patients referred (71 percent), the heart was discovered to be structurally normal. The proportion of patients with normal hearts referred by general practitioners and hospital doctors were 81 percent, and 68 percent, respectively (p less than 0.004). There was considerable variation in the pattern of referral between doctors working in different hospitals. CONCLUSION: New referrals to centres dealing with congenital cardiac malformations are increasing alarmingly, with the majority of the children referred having normal hearts. This increase in demand for specialist services has important implications for resources and training. PMID- 15831161 TI - An institutional review of the value of computed tomographic angiography in the diagnosis of congenital cardiac malformations. AB - The ultra-fast, thin-cut computerised tomographic angiogram is an efficient method to diagnose extracardiac lesions associated with congenital cardiac disease. For the purposes of this review, we evaluated various facets of the technique as used in 30 patients who were referred for diagnosis of congenital cardiac disease. The technique had high diagnostic accuracy, with a sensitivity of 93 percent in 15 of these patients referred for either interventional catheterisation or surgery. There were no immediate side-effects associated with the scanning procedure. The scan was also found to be more cost-effective as compared to an alternative noninvasive modality for imaging modality, namely magnetic resonance imaging. The angiographic technique, however, does expose the child to between 2 and 2.5 rems of radiation, despite the short period of scanning, of 10 plus or minus 2 seconds. PMID- 15831162 TI - Double aortic arch in a patient with Fallot's tetralogy. PMID- 15831163 TI - A surgically correctable cause of dilated cardiomyopathy in infancy. PMID- 15831164 TI - Current issues and perspectives in hypoplasia of the left heart. AB - Hypoplastic left heart syndrome is a rare but serious form of congenital cardiac disease, characterized by underdevelopment of the components of the left heart, rendering the left ventricle non-functional. Its aetiology is largely unknown, but there is certainly a genetic component. Prenatal diagnosis nowadays uncovers about half of cases. Postnatal options for treatment include comfort care, 3 stage palliative surgery, or cardiac transplantation. In this review, we discuss the morphology, possible pathogenetic mechanisms, clinical management, and perspectives of prenatal intervention based on work in animal models. PMID- 15831165 TI - Altered expression of dystrophin within the thoracic aorta in coarctation. AB - Although persisting endothelial dysfunction has been established in the vasculature of patients following surgical repair of coarctation, it is unknown whether there are alterations in the cytoskeleton of the aorta in such patients. We compared staining of N-terminus dystrophin in the smooth muscle of the aortic wall of a patient with coarctation to that in a patient without coarctation, the latter undergoing surgical treatment of a double aortic arch. There was a marked difference in the pattern of expression of dystrophin between the two, with the coarcted specimen demonstrating marked fragmentation but normal intensity of staining. As far as we are aware, ours is the first report to demonstrate the presence of dystrophin in the smooth muscle of the aorta. Alterations in the cytoskeletal structure may account for underlying aberrations in endothelial function in such patients, and is a topic that warrants further investigation. PMID- 15831166 TI - Spontaneous improvement of severe right ventricular dysfunction in the setting of hypoplasia of the left heart. AB - Right ventricular dysfunction is known to occur after the first stage of the Norwood sequence for treatment of patients with hypoplasia of the left heart. In a subset of patients, such ventricular dysfunction occurs without a specific anatomical cause. We describe two such cases with severe right ventricular dysfunction. In both cases, magnetic resonance imaging was used accurately to measure ventricular function and assess the arterial trunks. In both cases, cardiac transplantation was considered, but right ventricular function improved without invasive management. Transient right ventricular dysfunction in these cases may be due to the reduced ability of the right ventricle to adapt to the systemic vasculature. The improvement in ventricular function in these two cases may be due to delayed adaptation. PMID- 15831167 TI - A tunnel from the left sinus of Valsalva to the right atrium. AB - We describe an individualised approach in the rare situation of a tunnel between the left aortic sinus of Valsalva and the junction of the superior caval vein with the right atrium. It is necessary, in this rare anomaly, to identify the origin of the left coronary artery prior to intervention. PMID- 15831168 TI - Modification of the arterial switch procedure in the presence of a rare coronary arterial pattern. AB - Numerous techniques have been described for translocation of the coronary arteries in the setting of the arterial switch procedure. In the case we describe, the close alignment to the aortic and the pulmonary wall of the main stem supplying the left anterior descending and circumflex arteries, and dual origin of this vessel and the right coronary artery from sinus #2, required an individual approach. After taking down the main stem feeding the left anterior descending and circumflex arteries in the usual manner, we opted for a connection maintaining the artery in its existing site, and punched a hole into the neo aorta just in front of the coronary orifice. We then used a piece of glutaraldehyde-preserved pericardium as a hood, anastomosing the right coronary artery to the new aorta by means of the usual trap door technique. PMID- 15831169 TI - Anomalous drainage of the inferior caval vein to the left atrium. AB - We report a girl, aged 11(7/12) years, who presented with cyanosis. Cardiac catheterization showed occlusion of the infrahepatic segment of the inferior caval vein, with drainage of the hepatic veins into the left atrium. Transoesophageal echocardiography revealed an anomalous Eustachian valve that baffled the vein to the left atrium. This lesion is an extremely rare cause of cyanosis. PMID- 15831170 TI - The role of vasopressin in treating systemic inflammatory syndrome complicated by right ventricular failure. AB - Severe right ventricular failure complicated a postoperative systemic inflammatory response in a 33-year-old woman after surgical repair of congenital cardiac malformations. Volume loading, and high doses of catecholamines, failed to produce improvement, but treatment with vasopressin improved all haemodynamic parameters, and also allowed reduction of the other inotropes. After 10 days, the patient was discharged in stable condition from the intensive care unit. PMID- 15831171 TI - An experience suggesting an expanded role for the excimer laser catheter in neonates with obstructive lesions in the heart. AB - I have previously reported the successful use of the 0.9 millimetre excimer laser catheter in 2 neonates with pulmonary valvar atresia and intact ventricular septum. In this report, I describe the expanded role of the excimer laser in a premature infant weighing 1.8 kilograms who presented with an occluded right pulmonary artery secondary to an organized thrombus. Successful reconstruction of the right pulmonary artery was performed by sequential use of lasers of 0.9, 1.4, and 2 millimetres diameter, followed by conventional balloon angioplasty. As is now the situation in adults, laser catheter intervention should now be an integral part of the armamentarium of the paediatric cardiac interventionalist. PMID- 15831174 TI - Highlights of the meeting of the Psychosocial Working Group of the Association for European paediatric Cardiology, Vienna, March 4-6, 2004. PMID- 15831177 TI - Targeting the molecular basis for tumour hypoxia. AB - Tumour hypoxia stems from impaired oxygen delivery as a result of a disorganised tumour vasculature and inadequate blood supply. Hypoxic tumours are highly resistant to chemotherapy and radiation therapy and correlate with a poor patient prognosis. Hypoxia is a powerful stimulus for the expression of genes involved in cell survival and angiogenesis. A key factor in this process is hypoxia-inducible factor (HIF), which regulates transcription of hypoxia-activated genes. Efforts are currently under way to develop targeted cancer therapeutics to hypoxia activated pathways, and in particular to the transcription factor HIF. PMID- 15831178 TI - Roles for software technologies in advancing research and theory in educational psychology. AB - While reviews abound on theoretical topics in educational psychology, it is rare that we examine our field's instrumentation development, and what effects this has on educational psychology's evolution. To repair this gap, this paper investigates and reveals the implications of software technologies for researching and theorizing about core issues in educational psychology. From a set of approximately 1,500 articles published between 1999 and 2004, we sampled illustrative studies and organized them into four broad themes: (a) innovative ways to operationalize variables, (b) the changing nature of instructional interventions, (c) new fields of research in educational psychology, and (d) new constructs to be examined. In each area, we identify novel uses of these technologies and suggest how they may advance, and, in some instances, reshape theory and methodology. Overall, we demonstrate that software technologies hold significant potential to elaborate research in the field. PMID- 15831179 TI - Teachers' emotional expression about disruptive boys. AB - OBJECTIVES: To assess teachers' emotional expression about pupils using the Five Minute Speech Sample (FMSS) and coding procedures for parental expressed emotion (EE). To compare EE for disruptive and non-disruptive pupils. METHOD: Twenty-one teachers provided speech samples for both a disruptive and a non-disruptive pupil in their class selected using standard behaviour rating scales. RESULTS: Teachers' emotional expression was reliably measured using EE codings. Teachers displayed no emotional overinvolvement (EOI) and made few critical comments. High EE, characterized by criticism and a lack of positive comments, was associated with children's behavioural difficulties. Multiple regression suggested that conduct problems rather than hyperactivity were associated with high EE. DISCUSSION: Results support the application of certain elements of the EE construct to teachers' emotional expression about pupils. However, there was an absence of EOI and a lack of association between relationship and other EE categories. The absence of this association suggests that EE might be most usefully considered as a measure of teachers' emotional response to pupils, rather than the emotional quality of the teacher-pupil relationship. PMID- 15831180 TI - Patterns of language impairment and behaviour in boys excluded from school. AB - BACKGROUND: High levels of behaviour problems are found in children with language impairments, but less is known about the level and nature of language impairment in children with severe behavioural problems. In particular, previous data suggest that at primary age, receptive impairments are more closely related to behaviour problems, whereas expressive language has a closer link at a later age. AIMS: The study assessed expressive and receptive language problems in boys excluded from primary and secondary schools, to investigate the extent of impairment, the pattern of relations between age, receptive and expressive language, and relations with different aspects of behaviour. SAMPLE: Nineteen boys (8 - 16 years of age) who had been excluded from school and 19 non-excluded controls matched for age and school participated. METHOD: The sample was given assessments of: receptive language from the British Picture Vocabulary Scale (BPVS), and Wechsler Objective Language Dimensions (WOLD); expressive-language evaluations from the Wechsler Intelligence Scale for Children (WISC); auditory working memory evaluations from the Clinical Evaluation of Language Fundamentals (CELF); verbal reasoning (from the WISC); and non-verbal IQ assessments Raven's matrices. Teachers completed behaviour ratings using the Strengths and Difficulties Questionnaire (SDQ). RESULTS: Excluded boys were significantly poorer than controls on expressive measures but similar on receptive language and non-verbal IQ. Boys excluded from primary school were poorer than controls on auditory working memory. Expressive problems were linked with high levels of emotional symptoms. CONCLUSION: Many of the excluded boys had previously unidentified language problems, supporting the need for early recognition and assessment of language in boys with behaviour problems. Expressive problems in particular may be a risk factor. PMID- 15831181 TI - Activity in children with ADHD during waiting situations in the classroom: a pilot study. AB - BACKGROUND: According to the optimal stimulation theory and the delay aversion hypothesis, children with attention deficit hyperactivity disorder (ADHD) experience difficulties when they are confronted with low levels of stimulation and delay, respectively. AIM: This study investigated the activity level of children with ADHD during waiting situations in the classroom. Three series of hypothesis were made: (1) with respect to the comparison between waiting and non waiting intervals, (2) with respect to the effects of non-temporal stimulation, and (3) with respect to the effects of temporal stimulation on behaviour during waiting. SAMPLE AND METHOD: The activity level of 14 children with ADHD and 14 control children between the ages of 6 and 11 years was observed during two non waiting class situations and three waiting situations: without any stimulation, in the presence of nontemporal stimulation and in the presence of temporal stimulation. RESULTS: Both groups of children obtained higher activity scores for all behavioural dimensions during waiting compared with non-waiting situations. The results further revealed additive effects of waiting and diagnostic group on behaviour. Additional nontemporal stimulation during waiting affected the behaviour of all children for most behavioural characteristics. For noisiness, additive effects were also found for diagnostic group and either non-temporal stimulation or temporal stimulation. For restlessness, a trend for an interaction effect between diagnostic group and nontemporal stimulation was found. CONCLUSION: The findings have clear implications for school observations within an assessment protocol. PMID- 15831182 TI - The management of cognitive load during complex cognitive skill acquisition by means of computer-simulated problem solving. AB - This study compared the effects of two information presentation formats on learning to solve problems in electrical circuits. In one condition, the split source format, information relating to procedural aspects of the functioning of an electrical circuit was not integrated in a circuit diagram, while information in the integrated format condition was integrated in the circuit diagram. It was hypothesized that learners in the integrated format would achieve better test results than the learners in the split-source format. Equivalent-test problem and transfer-test problem performance were studied. Transfertest scores confirmed the hypothesis, though no differences were found on the equivalent-test scores. PMID- 15831183 TI - Knowledge accessibility, achievement goals, and memory strategy maintenance. AB - BACKGROUND: An important aim of educational psychology is to account for the difficulties in cognitive strategy maintenance. Possible explanations may be found in developmental studies concerning the interdependence of knowledge accessibility and strategy use, and in current achievement goal models which assume that individuals with a learning goal use deeper strategies than those with a performance goal. AIMS: The purpose of this study was to examine how knowledge accessibility and achievement goals separately or conjointly affect the maintenance of a categorization strategy, recall performance, and perception of the strategy (utility, difficulty, effort) in a free recall task. SAMPLE: Participants were 93 first-year students from a human science university in France. METHOD: In the first phase, the students were taught how to use a categorization strategy in a recall task involving highly accessible (typical) items. In a subsequent transfer task, the same students were assigned to one of four experimental conditions where they were asked to recall a typical or atypical list and where a learning or performance goal was induced. RESULTS: Analysis showed that whatever the typicality of the list, a learning goal promoted maintenance of the categorization strategy, and of perception of its utility. Performance goal participants only maintained this strategy and its perceptions on typical lists, while on atypical lists they display less clustering and found the strategy less useful. CONCLUSION: Results suggest that maintenance of a cognitive strategy is facilitated when there is compatibility between a performance goal and the task demands, or when the participants are learning-goal oriented. PMID- 15831184 TI - Pupils' evaluation and generation of evidence and explanation in argumentation. AB - BACKGROUND: Studies on argument have found that participants tend to prefer explanations to evidence. This apparent bias toward explanation has been qualified recently by research that has found it to diminish with the availability of evidence. AIM: This study examines the use of explanation versus evidence in the context of argumentation with reference to the goals of particular argument situations. SAMPLE: Seventy-nine eighth-grade pupils at a regular, urban middle school. METHOD: The pupils read argumentation scenarios, each having the stated goal of either explaining or proving a claim. The pupils rated the degree to which each of two provided assertions (one a theoretical explanation, and the other evidence-based) helped achieve the goal of the argument. On a second task, the pupils chose which of the two assertions should be more effective in achieving the argument goal. On the third task, the pupils generated either an explanation or evidence for each of the argumentation scenarios. RESULTS: Pupils demonstrated sensitivity to the relative epistemic strength of explanation and evidence. They rated explanations as more advantageous in achieving the explanation goal, and evidence as more advantageous in achieving the proof goal. Conversely, however, when asked to generate or recall an explanation or evidence, pupils produced more explanations than evidence independent of the argumentation goal. CONCLUSIONS: The study refines the definition of argumentation context to include specific goals. Pupils were sensitive to the context of the argumentation situation (e.g.goals, availability of evidence). However, they appeared to have a disposition toward explanation when asked to produce an explanation or evidence-based justification. PMID- 15831185 TI - Cognitive skills in mathematical problem solving in Grade 3. AB - BACKGROUND: Research on the relationship between cognitive skills and mathematical problem solving is usually conducted on adults or on participants with acquired deficits associated with brain injury (e.g.Cipolotti, 1995; Cohen, Dehaene, & Verstichel, 1994; McCloskey, 1992). AIMS: In these studies we wanted to make a contribution to the field of children's mathematical problem solving. The first aim of this study was to investigate whether mathematical problem solving in children is merely determined by semantic elaboration, as hypothesized in some of the models of adult processing (semantic hypothesis). In addition, we aimed to investigate whether there is a continuum from very good to very poor mathematical problem solving among children with mathematical learning disabilities showing immature cognitive skills (maturational lag hypothesis). SAMPLE: The participants were 376 third graders and 107 second graders. METHOD: The internal structure of the data was analysed with a principal components analysis. In addition, two MANOVA were conducted to compare children with learning disabilities or problems with age-matched and performance-matched subjects. RESULTS: Two components, a semantic and a non-semantic one, were needed to account for an adequate fit of the dataset. In addition, children with mathematical learning disabilities had less-developed cognitive skills compared with peers without learning disabilities, but they did not differ from younger children on seven of the nine cognitive skills. CONCLUSIONS: This study highlighted that children's mathematical problem solving is not determined by one general component. The picture is more complex, since two mathematics components were found. In addition, although our findings point in the direction of the maturational lag hypothesis it may be important to assess the different cognitive skills and especially assess the number system knowledge, since it seems below average in children with mathematical learning disabilities, compared with the knowledge of younger children with comparable skills in mathematics. PMID- 15831187 TI - Health care workforce planning: can it ever work? PMID- 15831188 TI - Evidence-based medicine and patient-centred medicine: the need to harmonize. PMID- 15831189 TI - US physicians' perceptions of the effect of practice guidelines and ability to provide high-quality care. AB - OBJECTIVES: In the USA, health care organizations frequently disseminate practice guidelines to physicians, but physicians often resist implementing guidelines when they perceive no improvements in quality of care will result. Greater involvement with a single health care organization may a inverted exclamation market physicians' perceptions of guidelines. We examined the relationship between the perceived effect of guidelines on practice and perceived quality of care for US primary care physicians (PCPs) and specialists with varying levels of financial involvement with a single managed care organization. METHODS: Data were from the 1996-1997 Community Tracking Study, a nationally representative, cross sectional survey of 12,528 physicians. Data were adjusted for possible confounders using ordinal logistic regression. RESULTS: Almost half the physicians described a moderate to very large perceived effect of guidelines (46% of PCPs, 46% of specialists). Physicians' financial involvement with a single organization was modest: PCPs received on average 24% of their revenue from their largest contract, while specialists averaged 18%. For specialists, increasing perceived effect of guidelines was associated with increasingly negative perceptions of quality of care [beta= -0.16, 95% confidence interval (-0.22, 0.10)]. Similar results were obtained for PCPs with low levels of financial involvement with a single organization. However, this negative association disappeared for PCPs with higher levels of financial involvement. CONCLUSIONS: PCPs with substantial financial involvement with a single organization who perceive greater effects of guidelines on practice have less negative perceptions of their ability to provide high-quality care. Although our data cannot confirm a causal relationship, financial involvement with a single organization may be one factor linking practice guidelines to high-quality care. PMID- 15831190 TI - Health status and health care of immigrants in Canada: a longitudinal analysis. AB - OBJECTIVES: This paper focuses upon health status, need for care, and use of health care from 1994/95 to 2000/01 in the Canadian foreign-born population. METHODS: Using Statistics Canada's longitudinal National Population Health Survey, descriptive and survival analyses are used to explore immigrant health status and health care. RESULTS: The health status of immigrants quickly declines after arrival, with a concomitant increase in use of health care services. However, survival analysis of the risk of a change to poor health indicates no difference between immigrants and the native-born. Similarly, there is no difference in the risk of hospital use between the two populations. CONCLUSIONS: The health status of recent immigrant arrivals is observed to decline towards that of the native-born population, while health care utilization increases. However, increased use may not be sufficient to offset declines in health, meaning that need for health care within the immigrant population may be unmet. PMID- 15831191 TI - Patients' and general practitioners' views of what constitutes appropriate hypertension management. AB - OBJECTIVES: To explore how patients and general practitioners (GPs) construct the concept of appropriateness in the context of hypertension management. METHOD: Q methodology was used. The study involved 120 patients and 12 GPs ranking 42 statements according to their degree of agreement or disagreement when considering appropriate hypertension management. The statements, comprising both clinical and non-clinical attributes, were developed from a qualitative study. Factor analysis of the data, using PQMethod computer software, determined if any patterns were discernible. RESULTS: Patients (n = 92) and GPs (n = 10) exclusively clustered to six factors (factor loadings > or = 0.5, P <0.01), which accounted for 77% of the total variance. The findings indicated that patients and GPs consider appropriate hypertension management in different ways. The GPs indicated that they considered non-pharmacological measures highly important, whereas 72% of patients were ambivalent. The patients clustered to five appropriateness factors, which varied in the degree of involvement patients had, or wished to have, in their hypertension management. Of these five, two were chosen by 73 patients. CONCLUSION: GPs' views differ from those of patients and there is variation between patients, which has important implications for patient centred care. Further application of Q-methodology to explore patients' views of appropriateness of other medical conditions would be valuable. PMID- 15831192 TI - Conceptual issues in the analysis of cost data within cluster randomized trials. AB - Cluster randomized controlled trials (RCTs) are increasingly used in economic evaluations of social, educational and health care interventions. Methodological research has, therefore, been spread across several disciplines, with the result that it has taken many years for guidelines on good statistical practice in the design and analysis of such trials to become easily accessible to health service researchers. These guidelines remain incomplete, however, because they do not take account of issues specific to the analysis of cost data. In particular, they fail to recognize that the calculation of confidence intervals around costs needed to inform health care priority setting raises unique methodological issues. If poorly designed trials are to be avoided in future (including those by the authors), then collaboration between triallists and health economists is required. This paper sets out a framework that should facilitate such collaboration and draws attention to problems that must be addressed quickly in the design of cluster-based economic evaluations. PMID- 15831193 TI - Contested ground: how should qualitative evidence inform the conduct of a community intervention trial? AB - This paper presents issues which arose in the conduct of qualitative evaluation research within a cluster-randomized, community-level, preventive intervention trial. The research involved the collection of narratives of practice regarding the intervention by community development officers working in eight communities over a two-year period. The community development officers were largely responsible for implementing the intervention. We discuss the challenges associated with the collection of data as the intervention unfolded, in particular, the disputes over cues to revise and adjust the intervention (i.e. to use the early data formatively). We explore the ethical uncertainties that arise when multiple parties have different views on the legitimacy of types of knowledge and the appropriate role of research and theory in various trial stages. These issues are discussed drawing on the fields of ethnography, community psychology, epidemiology, qualitative methodology and notions of research reflexivity. We conclude by arguing that, in addition to the usual practice of having an outcome data-monitoring committee, community intervention trials also require a process data-monitoring committee as a forum for debate and decision-making. Without such a forum, the relevance, ethics and position of qualitative evaluation research within randomized controlled trials are destined to be a point of contention rather than a source of insight. PMID- 15831194 TI - Effect of the hospital nursing environment on patient mortality: a systematic review. AB - OBJECTIVE: Research has examined the effect of the structure of health systems on health outcomes, but not how outcomes are affected by the nursing environments in hospitals. Our objective was to gather, critically appraise and synthesize all relevant primary research on the effect of the nursing environment on patient mortality. METHODS: Five electronic bibliographic databases were searched from their beginning through to May/June 2001, and Medline and CINAHL were updated to March 2004, using pre-determined search strategies and inclusion criteria. Studies were included if they met pre-determined criteria, reporting primary data both on a hospital environment and patient mortality. Methodological rigour was appraised using accepted criteria for the evaluation of research protocols, including case-mix adjustment. RESULTS: This paper focuses on 27 identified studies that investigated the impact of one or more attributes of the nursing environment on patient mortality. Nineteen studies found an association between one or more unfavourable attributes and higher mortality. There was considerable variability in attribute and outcome measures, settings and research quality across studies. This precluded statistical pooling of results. CONCLUSIONS: On balance, current evidence indicates that social and environmental attributes of hospital nursing practice have an effect on the outcomes of care. Before optimal practice settings can be designed, further research of greater rigour is needed to provide a better understanding of the mechanisms that link the nursing environment to patient outcomes. PMID- 15831195 TI - Evidence-based policy making in health care: what it is and what it isn't. AB - In this paper, I aim to re-establish the meaning and importance of the concept of 'evidence-based policy making' (EBP) in health care. The term EBP is often misunderstood as being either vacuous (who thinks that public policy should not be based on evidence?), unrealistic (the naive product of ivory tower thinking) or conservative (an excuse permanently to delay reform). It need be none of these things. EBP should be thought of as a set of rules and institutional arrangements designed to encourage transparent and balanced use of evidence in public policy making. As well as controlled trials and observational studies, a broad range of theoretical and empirical evidence about human behaviour may be relevant to predicting policy outcomes - including stakeholder opinions and other sources of intelligence that might not qualify as scientific research. Gradual progress towards EBP, properly understood, has the potential to facilitate open democracy and to improve policy outcomes. The argument is illustrated using examples based on large-scale policies of health care reform in England, where progress towards EBP over the last decade has been real but modest. PMID- 15831196 TI - Limits to medicine. Medical nemesis. PMID- 15831198 TI - The public uses health system report cards to make decisions about their health care. PMID- 15831199 TI - Did the invisible hand rock the cradle? PMID- 15831200 TI - Tumour targeting: biological factors and formulation advances in injectable lipid nanoparticles. AB - Cancer chemotherapeutic agents are often administered systemically. Following systemic administration, numerous biological factors associated with the tumours influence the delivery of the drugs to the tumours. These factors have been extensively studied for the last 2 decades. The influence of these biological factors has brought about a drastic change in the design of drug delivery systems to solid tumours. This review discusses the various biological factors influencing drug delivery to tumours and the subsequent development of injectable delivery systems (i.e., lipid-based nanoparticles (SLNs)) for adequate delivery of drug to solid tumours. PMID- 15831201 TI - Dermal delivery of desmopressin acetate using colloidal carrier systems. AB - Recently, the transdermal route has received attention as a promising means to enhance the delivery of drug molecules, particularly peptides, across the skin. In this work, the skin penetration profiles of desmopressin acetate from a colloidal system (water-in-oil microemulsion) and an amphiphilic cream, a standard formulation, were determined using Franz diffusion cells and compared. In the case of the microemulsion, the total percentages of dose obtained from different skin layers(stratum corneum to subcutaneous tissue) were 3.30+/-0.67, 7.37+/-2.43 and 15.54+/-2.72 at 30, 100 and 300 min, respectively. Similarly, 5.19+/-0.96, 8.04+/-0.97 and 14.4+/-5.15% of the dose applied was extracted from the skin treated with the cream. About 6% of the applied dose reached the acceptor compartment from the microemulsion instead of 2% from the cream within 300 min. The concentration of drug that penetrated into the upper layers of the skin was higher from the cream than from the microemulsion at all time intervals. On the other hand, a higher amount of drug was found in the deeper skin layers and in the acceptor compartment from the microemulsion. PMID- 15831202 TI - Influence of magnesium aluminium silicate on rheological, release and permeation characteristics of diclofenac sodium aqueous gels in-vitro. AB - The effect of magnesium aluminium silicate (MAS) on rheological, release and permeation characteristics of diclofenac sodium (DS) aqueous gels was investigated. DS aqueous gels were prepared using various gelling agents, such as 15% w/w poloxamer 407 (PM407), 1% w/w hydroxypropylmethylcellulose (HPMC), and 1% w/w high and low viscosity grades of sodium alginate (HV-SA and LV-SA, respectively). Different amounts of MAS (0.5, 1.0 and 1.5% w/w) were incorporated into the DS gels. Incorporation of MAS into the DS gels prepared using SA or PM407 caused a statistical increase in viscosity (P<0.05) and a shift from Newtonian flow to pseudoplastic flow with thixotropic property. The DS release rates of these composite gels were significantly decreased (P<0.05) when compared with the control gels. This was due to an interaction between MAS and PM407 or SA, and adsorption of DS onto MAS particles. Moreover, a longer lag time and no change in DS permeation flux were found when MAS was added to the gels. The findings suggest that the rheological characteristics of gels prepared using PM407 or SA could be improved by incorporating MAS. However, the use of MAS could retard the DS release and extend the lag time of DS permeation. PMID- 15831203 TI - Experimental modelling of drug membrane permeability by capillary electrophoresis using liposomes, micelles and microemulsions. AB - Capillary electrophoresis (CE) was evaluated as an in-vitro format for experimental modelling of membrane permeability using only nanogram quantities of drug compounds. The rationale for the CE technique emanates from emulation of a lipid-like pseudo-stationary phase that governs separations mainly as a result of differences in molecular size, lipophilicity, hydrogen bonding and charge, all of which also have a strong influence on in-vivo drug absorption. By means of micellar, microemulsion and liposome electrolytes, the migration behaviour was studied at 37 degrees C for 22 model drug compounds. The generated CE retention factor data were then compared with membrane permeability reference data. Both simple log D and more common Caco-2 cell parameters were evaluated. In addition, permeation through intestinal segments of rat ileum and rat colon was included. An improved correlation was obtained in the order: micellar < microemulsion < liposome systems. Although the correlation for the best liposome CE system was only R(2)=0.77, the evaluation results for all emphasized the strength and flexibility of CE for assessing specific drug-membrane interaction through tailor made lipophilic media. PMID- 15831204 TI - Effect of enzyme inducers and inhibitors on the pharmacokinetics of oltipraz in rats. AB - A series of in-vitro and in-vivo experiments, using various inducers and inhibitors of hepatic microsomal cytochrome P450 (CYP) isozymes, was conducted to study oltipraz pharmacokinetics in rats. In in-vivo studies, oltipraz at a dose of 10 mg kg(-) was administered intravenously to rats. In rats pretreated with SKF 525-A (a nonspecific CYP isozyme inhibitor in rats; n-9), the time-averaged total body clearance (CL) of oltipraz was significantly slower (56.6% decrease) than that in untreated rats (n=9). This indicated that oltipraz is metabolized via CYP isozymes in rats. Hence, various enzyme inducers or inhibitors were used in in-vitro and in-vivo studies in rats. In rats pretreated with 3 methylcholanthrene (n=9 and 8 for untreated and treated groups, respectively), phenobarbital (n=7 and 10 for untreated and treated groups, respectively) or dexamethasone (n=7 and 12 for untreated and treated groups, respectively) (main inducers of CYP1A1/2, 2B1/2 and 3A1/2 in rats, respectively), the CL values were significantly faster (38.4, 94.4 and 33.6% increase, respectively). In rats pretreated with sulfaphenazole (n=8 and 9 for untreated and treated groups, respectively), quinine (n=7 and 9 for untreated and treated groups, respectively) or troleandomycin (n=8 and 9 for untreated and treated groups, respectively) (main inhibitors of CYP2C11, 2D1 and 3A1/2 in rats, respectively), the CL values were significantly slower (31.0, 27.6 and 36.3% decrease, respectively). The in vivo results with various enzyme inhibitors correlated well with the in-vitro intrinsic clearance for disappearance of oltipraz (CL(int)) (n=5, each). The above data suggested that oltipraz could be metabolized in male rats mainly via CYP1A1/2, 2B1/2, 2C11, 3A1/2 and 2D1. PMID- 15831205 TI - Investigation of oral bioavailability and brain distribution of the Ind(8)-Val conjugate of indinavir in rodents. AB - Protease inhibitors are successfully used for the treatment of acquired immune deficiency syndrome (AIDS) although their biopharmaceutical characteristics are not optimal. Prodrugs have therefore been synthesized to increase protease inhibitor bioavailability and brain distribution. Among several compounds tested, a valine derivative of indinavir (Ind(8)-Val) showed promising characteristics using an in-vitro Caco-2 cell model. The objective of this study was to further investigate this compound using in-situ and in-vivo approaches. The pharmacokinetics of indinavir (Ind) and Ind(8)-Val were investigated in rats after intravenous and oral administration. Free indinavir resulting from in-vivo hydrolysis of Ind(8)-Val could not be detected in the plasma of rats receiving Ind(8)-Val. Furthermore Ind(8)-Val bioavailability was only 32% on average compared with 76% for indinavir, and effective permeability coefficients determined with a single-pass intestinal perfusion method were close to 25x10(6)cms(-1) for the two compounds. Brain-to-plasma concentration ratios in the post equilibrium phase after intravenous administration to mice were 9.7+/ 8.1% for indinavir and 2.5+/-2.7% for Ind(8)-Val. In conclusion, the promising biopharmaceutical characteristics of Ind(8)-Val suggested from previous in-vitro experiments with the Caco-2 cell model were not confirmed by in-situ and in-vivo experiments. PMID- 15831206 TI - Effect of flavones on rat brain and lung matrix metalloproteinase activity measured by film in-situ zymography. AB - We have evaluated the inhibitory activity of flavone, nobiletin, and heptamethoxyflavone on matrix metalloproteinase (MMP) activity in the rat. MMP in 9000-g supernatant fraction of lung homogenate was activated by p-aminophenyl mercuric acetate (APMA), and gelatinolytic activity was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Coomassie staining. This activity should be related to MMP-2 and/or MMP-9 and was confirmed by gelatin zymography. Fluorescent-conjugated collagen used as a substrate for collagenolytic activity wasinvestigated by SDS-PAGE also. The film in-situ zymography method was applied to rat brain and lung tissue in the same manner. Flavone and nobiletin inhibited the APMA-stimulated gelatinolytic activity and also the collagenolytic activity by more than 75%. The film in-situ zymography method indicated that these compounds might be potent inhibitors of MMP, suggesting the specific inhibition of localized MMP in brain hippocampus and/or lung terminal bronchioles, which may contribute to the prevention of some types of brain disease or cancer invasion and metastasis. PMID- 15831207 TI - Terpinen-4-ol: mechanisms of relaxation on rabbit duodenum. AB - The effect of terpinen-4-ol was studied on rabbit duodenum in-vitro. Terpinen-4 ol induced relaxation of the basal tonus (IC50 170.2 (95% confidence interval, 175-204) microM) with a maximal relaxant response of 180.4+/-3.9% (n=6) of the contraction induced by 60 mM [K(+)]. The maximal relaxation induced in control conditions was not affected (P>0.05) by pretreatment of the tissues with phentolamine (50 microM) or propranolol (10 microM), N(G) nitro-L-arginine methyl ester (L-NAME; 1 mM), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 100 microM), hexamethonium (1 mM), tetrodotoxin (1 microM), the mixture charybdotoxin apamin (1 microM), glibenclamide (10 microM), 4-aminopyridine (10 microM) or tetraethyl-ammonium (100 microM). In addition, terpinen-4-ol completely relaxed tissues precontracted with 60 mM [K(+)] solutions (IC50 325.9 (245.1-433.1) microM) and also blocked (IC50 154.7 (117.7-191.7) microM) the phasic component of this contraction. At a concentration of 195 and 650 muM it reduced by 41.3+/ 3.4% and 75.4+/-3.1%, respectively the maximal contractile response to Ca(2+) in depolarized duodenum. Terpinen-4-ol completely blocked the component of carbachol induced contraction, which was resistant to nifedipine (100 microM) pretreatment or to a Ca(2+)-free solution. These data show that terpinen-4-ol relaxes intestinal smooth muscle and suggest that this effect is myogenic in nature and depends on calcium antagonism. PMID- 15831209 TI - Nicardipine, a calcium antagonist, does not aggravate intracerebral haemorrhage in an intracerebral haemorrhage model in rats. AB - Despite controversy over their safety in patients with intracerebral haemorrhage, calcium antagonists are widely used in the treatment of hypertensive emergencies. Here, we investigated the effects of nicardipine on haematoma size and neurological deficit in a rat model of collagenase-induced intracerebral haemorrhage. Injection of collagenase (0.014 U) into the striatum induced haematoma (19.9+/-3.4 mm(3)) in the striatum and brain oedema. Drugs were infused from 30 min after collagenase injection for 3 h under conscious conditions. Nicardipine intravenously at 0.1, 1 and 10 microg kg(-1) min(-1) affected neither haematoma size nor the degree of brain oedema. Nicardipine at these doses provided a stable and dose-dependent decrease in mean blood pressure of 6%, 13% and 33%, respectively, with an increase in heart rate that was apparently caused reflexively. Further, nicardipine did not aggravate the neurological deficits in these intracerebral haemorrhage rats, primarily forearm flexion behaviour on suspension by the tail and circling behaviour. These results indicate that nicardipine infusion stably decreased blood pressure without affecting intracerebral haemorrhage in an intracerebral haemorrhage model in rats. PMID- 15831208 TI - Hydroquinone modulates reactivity of peroxynitrite and nitric oxide production. AB - Peroxynitrite (ONOO(-)), a potent cytotoxic oxidant formed by the reaction of nitric oxide ((.-)NO) and superoxide radical ((.-)O(2)(-)), may be rapidly lethal in a cellular milieu due to oxidization and nitration processes. In the present study, hydroquinone displayed strong ONOO(-) scavenging activity and inhibitory effect on NO production in murine macrophage RAW264.7 cells. Hydroquinone strongly scavenged ONOO(-)induced dihydrorhodamine 123 oxidation in a dose dependent manner compared with other reactive species such as (.-)O(2)(-) and (. )NO. Hydroquinone also decreased levels of ONOO(-) induced nitrotyrosine of glutathione reductase and consequently prevented the enzyme from ONOO(-) induced damage. Furthermore, hydroquinone suppressed NO production, a cellular pathway for ONOO(-) formation, in lipopolysaccharide-activated RAW264.7 cells via inhibition of inducible NO synthase expression. The inhibitory effect by hydroquinone seems to be mediated by interruption of lipopolysaccharide-induced signalling such as activation of nuclear factor-kappaB and extracellular signalrelated kinases 1 and 2. The results suggest that hydroquinone may potently modulate reactivity of ONOO(-) and may therefore be a useful agent against ONOO( ) mediated diseases. PMID- 15831210 TI - Mechanism of cell death induced by cationic dendrimers in RAW 264.7 murine macrophage-like cells. AB - Cationic dendrimers possess attractive nano-sized architectures that make them suitable as targeted drug/gene delivery systems. However, very little is known about their mechanisms of cell death in cellular systems. In the current study, the apoptotic and necrotic effects of starburst polyamidoamine(PAMAM) and polypropylenimine (DAB) dendrimers in cultured RAW 264.7 murine macrophage-like cells were investigated. Cationic dendrimer treatment produced a typically dose dependent cytotoxic effect on macrophage cells. RAW 264.7 cells exposed to cationic dendrimers exhibited morphological features of apoptosis. Apoptotic ladders were observed in DNA extracted from RAW 264.7 cells treated with cationic dendrimers. Analysis from flow cytometry demonstrated an increase in hypodiploid DNA population (sub-G1) and a simultaneous decrease in diploid DNA content, indicating that DNA cleavage occurred after exposure of the cells to cationic dendrimers. Also, cells treated with DAB dendrimer induced a higher percentage of sub-G1 population than those treated with PAMAM dendrimer at the same dose. In addition, it was shown that pre-treatment of RAW 264.7 cells with the general caspase inhibitor zVAD-fmk prevented some degree of apoptosis induced by cationic dendrimers, suggesting that apoptosis in macrophage cells involves a caspase dependent pathway. Macrophage cells were also found to be sensitive to induction of apoptosis by dendrimers, whereas NIH/3T3 cells (mouse fibroblast) and BNL CL.2 (mouse liver) cells did not undergo apoptosis. These results could be helpful for optimizing the biocompatibility of dendrimers used for targeted drug/gene delivery. PMID- 15831211 TI - Effect of aqueous Enicostemma littorale Blume extract on key carbohydrate metabolic enzymes, lipid peroxides and antioxidants in alloxan-induced diabetic rats. AB - The present study investigates the effect of oral administration of an aqueous Enicostemma littorale whole plant extract on some key carbohydrate metabolic enzymes and antioxidant defence in alloxan-induced diabetes in rats. Rats were rendered diabetic by alloxan (150 mgkg(-1) body weight) administration. Oral administration of E. littorale extract for 45 days increased the activity of hexokinase and decreased the activities of glucose 6-phosphatase and fructose 1,6 bisphosphatase significantly in the serum, liver and kidney of diabetic rats. The extract lowered the concentration of thiobarbituric acid reactive substances and lipid hydroperoxides significantly in brain and increased it significantly in heart in diabetic rats. E. littorale administration increased the concentration of reduced glutathione and the activity of glutathione peroxidase in diabetic rats. The activities of superoxide dismutase and catalase were increased significantly by E. littorale treatment in diabetic rats. The effect of a 2 g kg( 1) dose was greater than that of a 1 gkg(-1) dose. Insulin (6 units kg(-1)) normalized all the parameters in diabetic rats. Our study has provided evidence for the antidiabetic activity of E. littorale aqueous extract. This study can also be extrapolated to clinical studies in future. PMID- 15831212 TI - Citrus extract modulates genotoxicity induced by cyclophosphamide in mice bone marrow cells. AB - The protective effect of citrus extract was investigated by using the micronucleus assay for anticlastogenic activity in mouse bone marrow cells; liver glutathione (GSH) content was determined against toxicity induced by cyclophosphamide. Mice were orally (gavage) pretreated with solutions of citrus peel extract (Citrus aurantium var. amara) prepared at three different doses (100, 200 and 400 mg kg(-1;) body weight) for 7 consecutive days. Then mice were injected intraperitoneally on the seventh day with cyclophosphamide (50 mg kg( 1)) and after 24 h killed for the evaluation of micronucleated polychromatic erythrocytes (MnPCEs) in bone marrow cells. Non-protein thiol levels in liver were estimated in mice injected with citrus extract with or without cyclophosphamide treatment. Administration of citrus extract before cyclophosphamide treatment significantly reduced the frequency of MnPCEs in mice bone marrow compared with the group treated with cyclophosphamide alone (P<0.0001 0.05). Citrus extract at a dose of 400 mg kg(-1) reduced MnPCEs 2.8 fold against genotoxicity induced by cyclophosphamide. Administration of cyclophosphamide depleted the GSH level in liver. Citrus extract showed excellent scavenging effects on 1,1-diphenyl-2-picryl hydrazyl radical (DPPH) at a concentration of 1.6 mg mL(-1). Application of citrus extract 1 h before cyclophosphamide treatment allowed GSH content to reach the normal level. It appeared that citrus extract, particularly flavonoids constituents with antioxidative activity, may return the GSH level to normal in stress conditions and reduces genotoxicity induced by cyclophosphamide in bone marrow cells. PMID- 15831213 TI - Nifedipine enhances cGMP production through the activation of soluble guanylyl cyclase in rat ventricular papillary muscle. AB - It is known that nifedipine, an L-type calcium channel blocker, increases cGMP production, which partially contributes to the relaxation of vascular smooth muscle. The aim of our investigation was to clarify whether or not nifedipine regulates cGMP production, which has a physiological role in cardiac muscle. To measure contractile responses and tissue cGMP levels, left ventricular papillary muscles prepared from male Wistar rats (350-400 g) were mounted in the isolated organ chamber under isometric conditions and electrically paced by means of platinum punctate electrodes (1 Hz, 1 ms duration). In papillary muscle preparation, the negative inotropic effect induced by nifedipine (30 to 300 nM) was significantly inhibited in the presence of ODQ(1H-[1,2,4]oxidazolo[4,3 a]quinoxaline1-one; 10 microM), a soluble guanylyl cyclase inhibitor. Furthermore, nifedipine (100 nM) strongly increased the tissue cGMP level, which was significantly decreased in the presence of ODQ. On the other hand,N(G) monomethyl-(L)-arginine (100 microM), a nitric oxide synthase inhibitor, did not inhibit either the negative inotropic effect or cGMP production induced by nifedipine. These results indicate that in rat left ventricular papillary muscle, nifedipine augments its negative inotropic effect at least partly through direct activation of cardiac soluble guanylyl cyclase but not nitric oxide synthase. PMID- 15831214 TI - Selective beta(1)-adrenoreceptor blocking activity of newly synthesized acyl amino-substituted aryloxypropanolamine derivatives, DPJ 955 and DPJ 890, in rats. AB - The in-vivo beta-adrenoreceptor antagonistic activity of test compounds DPJ 955 and DPJ 890 was assessed against beta-adrenoreceptor agonist (isoprenaline) induced tachycardia in anaesthetized rats. The selectivity to block isoprenaline responses on different &beta-adrenoreceptor subtypes (beta(1), beta(2) and beta(3)) of the test compounds was carried out on isolated rat right atria, isolated rat uterus and isolated rat colon preparations, respectively. Intravenous injection of isoprenaline alone in anaesthetized rats caused hypotension and tachycardia. DPJ 955 or DPJ 890 alone produced a fall in mean arterial pressure and bradycardia in a dose-dependent manner. Administration of isoprenaline to anaesthetized rats pre-treated with test compounds significantly blocked both the tachycardial and hypotensive responses induced by isoprenaline. The test compounds shifted the concentration response curves of isoprenaline towards the right for isolated rat right atrial preparations, rat uterus and rat colon, indicating beta(1), beta(2) and beta(3) adrenoreceptor blockade, respectively. The selectivity ratio for beta(1)/beta-adrenoreceptors to DPJ 955 and DPJ 890 was 64.6 and 83.2, respectively. DPJ 890 was more potent in blocking beta(1)-adrenoreceptors and was more selective towards beta(1) receptors than to other beta-adrenoreceptor subtypes. In conclusion, DPJ 955 and DPJ890 have beta adrenoreceptor blocking activity with high selectivity for the beta(1) adrenoreceptor subtype. PMID- 15831215 TI - High-speed digital imaging method for ciliary beat frequency measurement. AB - The aim of this study was to develop a high-speed digital imaging system and related software for ciliary beat frequency (CFB) analysis in order to establish an automated and reliable method that is observer independent and faster compared to the conventional computerized microscope photometry method. Using primary human nasal epithelial cell cultures, the CBF was recorded with a computerized microscope photometry system and a high-speed digital imaging system. To obtain a wide range of frequencies, glycocholate (0.5%) and chlorocresol (0.005%) were used as ciliostimulatory and cilio-inhibitory reference compounds, respectively. The mean values in hertz (+/- s.d.) obtained with the photometry and high-speed digital imaging systems were: controls 8.2 +/- 0.9 and 7.9 +/- 1.1; chlorocresol 5.0 +/- 0.9 and 5.1 +/- 1.1; glycocholate 9.8 +/- 1.0 and 9.7 +/- 0.8. A similar increase (by 20 and 24%) and decrease (by 38 and 35%) in CBF was determined by the two methods after glycocholate and chlorocresol treatment, respectively. The mean difference between the photometry and high-speed digital imaging methods was 0.2 +/- 0.6 Hz, and the Bland-Altman limits of agreement were from -1.0 to +1.4 Hz, suggesting that the results obtained by these two methods could be used interchangeably. These results show the reliability of the high-speed digital imaging system and the software developed for in-vitro CBF measurements. The advantages of the system include: (i) fast data acquisition and calculation, (ii) whole field automated CBF analysis and (iii) reduction in selection bias. PMID- 15831216 TI - Development of simple thiol-reactive liposome formulations, one-step analysis and physicochemical characterization. AB - The aim of this study was to synthesize simple thiol-reactive conjugates from maleimide and lipoamines (stearylamine or oleylamine) and to develop a simple, fast and low-cost method for the preparation of lyophilized general-purpose thiol reactive liposomes. A formulation of egg phosphatidylcholine dipalmitoylphoshatidylglycerol (9:0.1 molar ratio) was developed and characterized. Freeze-drying methodology was established to produce a stock of liposomes and the physicochemical characteristics of the reconstituted liposomes were compared with those of the initial preparation. The physicochemical properties (size and zeta potential) of the new liposomal formulations were studied. High-performance thin-layer chromatography coupled to a flame ionization detector was applied for one-step analysis of the liposomal components and for determining the maleimide-lipoamine conjugates phospholipid molar ratio. The differences concerning the incorporation efficiency of the synthetic conjugates into liposomes were discussed on the basis of their conformational properties. The small difference in structure between the two thiol-reactive conjugates (i.e., the C18 alkyl chain double bond) causes a considerable difference in phospholipids packing of the resulting lipidic bilayers of the liposomes; the conformational bending of conjugate maleimide-oleylamine may contribute to the final architecture of liposomes. PMID- 15831218 TI - The physiologic effects of laparoscopy: applications in oncology. AB - The use of laparoscopic approaches to surgical disease continues to advance quickly. Laparoscopy applied to oncologic surgery continues to be debated. We review the experience of laparoscopy as it relates to surgery for tumors. Specifically, we discuss the physiologic changes and tumor response to laparoscopy, as well as the current concepts explaining port site recurrence. PMID- 15831219 TI - Minimally invasive treatment of esophageal cancer: laparoscopic staging to robotic esophagectomy. AB - Minimally invasive surgical (MIS) procedures have become commonplace in modern surgical practice. The term minimally invasive surgery has been and continues to be interchangeably applied to describe laparoscopic, laparoscopic-assisted, thoracoscopic, and telesurgical (robotic) procedures. Minimally invasive surgical procedures for the treatment of benign and malignant disorders of the esophagus are being developed, refined, and clinically applied in parallel with the exponential availability of novel technologies and instrumentation. Herein, we review the progression from laparoscopic/thoracoscopic esophagectomy to telesurgical esophagectomy, presently termed minimally invasive esophagectomy, and describe the telesurgical procedure as well as early the clinical outcome experience. PMID- 15831220 TI - Minimally invasive treatment of stomach cancer. AB - The rate of detection of early gastric cancer has increased because of the development of diagnostic techniques, such as endoscopy, biopsy, and endoscopic ultrasonography. Recently, minimally invasive surgical procedures for benign gastric conditions have been advocated, and the laparoscopic approach is noted as a technique that increases the quality of life. However, the development of laparoscopic gastric resections and laparoscopically assisted gastric operations for malignancy still deserve a word of caution. Laparoscopic local resection of the stomach is used to treat mucosal cancer without lymph node metastasis, and laparoscopy-assisted distal gastrectomy is used to treat early gastric cancer with lymph node metastasis in the perigastric portion. According to short-term results reported by a small group of surgeons, laparoscopic approaches for gastric cancer result in a minimally invasive approach, early recovery, and decreased morbidity and mortality. However, the longterm results of these less invasive treatments are not known in advanced gastric cancer. If the results of randomized controlled studies for advanced gastric cancer are confirmed, the use of these techniques will spread worldwide and may become a standard technique for the resection of gastric cancer. PMID- 15831221 TI - Minimally invasive treatment of colon cancer. AB - The advantages of laparoscopy in the treatment of benign diseases have been well demonstrated. Compared with laparotomy, the laparoscopic approach is associated with a shorter hospitalization period, shorter duration of ileus, decreased postoperative pain, earlier return to work, and improved cosmesis. The role of laparoscopy for the treatment of gastrointestinal malignancy has had a slower evolution and been the subject of considerable debate over the past decade. Since 1991, several concerns have limited the widespread use of laparoscopy for attempted cure of colorectal carcinoma. This review aims to analyze the results of several studies published to date on short and long term outcome of laparoscopy for colorectal carcinoma, based on levels of evidence. From the least to the most convincing data, the hierarchy of study designs progresses through a spectrum ranging from retrospective reviews to prospective series, to case controlled, cohort, and ultimately randomized controlled trials. PMID- 15831222 TI - Splenic malignancy: a minimally invasive approach. AB - Since the advent of laparoscopy and its general acceptance for treating benign diseases, indications for malignant disease have been investigated. Recently, greater evidence shows that laparoscopy for malignant disease is oncologically safe. DESIGN: We review a minimally invasive approach to splenic malignancy and the common malignant diseases involving the spleen. We outline our preferred technique for splenectomy in detail. Additionally, the recent literature is reviewed regarding outcome after laparoscopic splenectomy for benign and malignant disease. The data from three studies, containing a total of 327 were analyzed. Complication rates, mortality, and length of stay were compared. RESULTS: There was no statistically significant difference identified between those undergoing laparoscopic splenectomy for benign versus malignant disease in terms of length of stay, complication rate or mortality. There were significant differences between the two groups in terms of operative time and spleen weight. DISCUSSION: In open splenectomy series for patients with malignant diseases of the spleen, complication and mortality are much higher when compared to those patients undergoing open splenectomy for benign disease. The discussed series show no difference in endpoints when laparoscopy is used. Laparoscopic splenectomy for malignant disease confers significant benefit and rapid recovery for an otherwise at risk population. PMID- 15831223 TI - New approaches to the minimally invasive treatment of pancreatic cancer. AB - Advances in minimally invasive surgery have revolutionized the field of surgery. Despite the great strides in equipment and experience, operative conduct remains confined by the limits of exposure. Retroperitoneal fat can be abundant and can contribute greatly to difficulty in exposure. Visceral organs ventral to the retroperitoneum preclude direct access and require optimal patient positioning to operate. Additionally, the major vascular pedicles all originate in the retroperitoneum off of the abdominal aorta or enter the inferior vena cava. The pancreas, in particular, is surrounded by the portal vein, celiac axis, superior mesenteric vein and artery, and splenic vein and artery. If injured during surgery, these vessels can present a life-threatening emergency. The issues related to the vasculature, coupled with the difficulty in resecting portions of the pancreas and the relative paucity of pancreatic procedures, have greatly concentrated these cases at tertiary care centers staffed by experienced laparoscopists. However, as surgical technology improves and fellowships train more surgeons with advanced laparoscopic skills, minimally invasive pancreatic surgery may diffuse with more community-based health care networks. PMID- 15831224 TI - New approaches to the minimally invasive treatment of liver cancer. AB - Unlike laparoscopic cholecystectomy, laparoscopic hepatectomy has been slow to gain acceptance because of its association with technical difficulties. Many surgeons feel there are few advantages in laparoscopic hepatectomy when compared to open surgery. The liver is the organ most susceptible to bleeding while dissecting the parenchyma and the resected liver usually requires a wide abdominal incision to deliver the resected specimen. Both the improvement of surgeons' skills and the development of technology have improved results, however, the indication of laparoscopic hepatectomy for malignancy is still controversial. This article focuses on the current status of minimally invasive treatment for liver malignancy. PMID- 15831225 TI - New approaches to the minimally invasive treatment of kidney tumors. AB - The incidence of renal cortical neoplasms has dramatically increased with the widespread use of abdominal imaging over the past 20 years. Coincidentally, the proportion of tumors that are smaller and incidentally detected has risen as well, indicative of a stage migration. The widespread application of minimally invasive and laparoscopic techniques to other organ systems has spurred the development of minimally invasive approaches to the management of renal tumors. The available data regarding laparoscopic nephrectomy, laparoscopic partial nephrectomy, and tissue ablative techniques, such as renal cryoablation, radiofrequency ablation, and high-intensity focused ultrasound are reviewed. PMID- 15831226 TI - New approaches to the minimally invasive treatment of adrenal lesions. AB - The advancement of laparoscopic adrenalectomy over the past decade has completely changed the surgical approach to adrenal tumors. As the incidence of incidentally discovered adrenal tumors increases, most patients with resectable lesions can undergo resection laparoscopically with minimal morbidity, shorter hospitalization, and low mortality. The spectrum of surgical approaches now available make it possible to provide an appropriate resection that is matched to the specific characteristics of each tumor. Experienced surgeons now resect some malignant tumors laparoscopically, with the option to convert to a hand-assisted or traditional open approach. PMID- 15831227 TI - New approaches to the minimally invasive treatment of lung cancer. AB - VATS lobectomies have now been performed for 12 years since the first VATS lobectomy. Although some controversies remain regarding the safety, morbidity, and mortality of that procedure, the procedure is clearly gaining momentum. The purpose of this paper is to review the current literature about the procedure. MATERIALS AND METHODS: Between 1992-2004, we performed 1100 VATS lobectomies in 595 (54.1%) women and 505 men (45.9%), mean age=71.2 years. Diagnoses were as follows: benign disease (53), pulmonary met (27), Lymphoma (5) and lung cancer (1015). 641 (63.1%) of the primary lung cancers were adenocarcinoma. With visualization on a monitor, anatomic hilar dissection and lymph node sampling or dissection were performed, primarily through a 5 cm incision without spreading the ribs. RESULTS: There were 9 deaths (0.8%)-none were intra-operative or due to bleeding. 932 patients had no postoperative complications (84.7%). Blood transfusion was required in 45/1100 (4.1%). Length of stay was median 3 days, mean 4.78 days. 180 patients were discharged on POD 1 or 2 (20%). Conversion to a thoracotomy occurred in 28 patients (2.5%). 5 patients developed recurrence in the incisions (0.57%). In 2003, 89% of 224 lobectomies were performed with VATS. CONCLUSIONS: VATS lobectomy with anatomic dissection can be performed with low morbidity and mortality. The risk of intraoperative bleeding or recurrence in an incision seems minimal. PMID- 15831228 TI - Ablative approaches to the minimally invasive treatment of breast cancer. AB - With the improvements in imaging techniques that have allowed the earlier detection of smaller breast cancers and the desire for improvements in cosmetic outcome, a number of minimally invasive techniques for the treatment of early stage breast cancers are being investigated. Ablative therapies, including laser ablation, focused ultrasound, microwave ablation, radiofrequency ablation, and cryoablation, have been described. All of these techniques have shown promise in the treatment of small cancers of the breast; however, additional research is needed to determine the efficacy of these techniques when they are used as the sole therapy and to determine the long-term local recurrence rates and survival associated with these treatment strategies. PMID- 15831230 TI - Endogenous inhibitors of angiogenesis in malignant gliomas: nature's antiangiogenic therapy. AB - Angiogenesis is necessary for tumor growth beyond a volume of approximately 2 mm(3). This observation, along with the accessibility of tumor vessels to therapeutic targeting, has resulted in a research focus on inhibitors of angiogenesis. A number of endogenous inhibitors of angiogenesis are found in the body. Some of these are synthesized by specific cells in different organs, and others are created by extracellular proteolytic cleavage of plasma-derived or extracellular matrix-localized proteins. In this review, we focus on angiostatin, endostatin, PEX, pigment epithelial-derived factor, and thrombospondin (TSP)-1 and -2, either because these molecules are expressed in malignant glioma biopsies or because animal studies in malignant glioma models have suggested that their therapeutic administration could be efficacious. We review the known mechanisms of action, potential receptors, expression in glioma biopsy samples, and studies testing their potential therapeutic efficacy in animal models of malignant glioma. Two conclusions can be made regarding the mechanisms of action of these inhibitors: (1) Several of these inhibitors appear to mediate their antiangiogenic effect through multiple protein-protein interactions that inhibit the function of proangiogenic molecules rather than through a specific receptor mediated signaling event, and (2) TSP-1 and TSP-2 appear to mediate their antiangiogenic effect, at least in part, through a specific receptor, CD36, which initiates the antiangiogenic signal. Although not proven in gliomas, evidence suggests that expression of specific endogenous inhibitors of angiogenesis in certain organs may be part of a host antitumor response. The studies reviewed here suggest that new antiangiogenic therapies for malignant gliomas offer exciting promise as nontoxic, growth-inhibitory agents. PMID- 15831231 TI - The role of interleukin-8 and its receptors in gliomagenesis and tumoral angiogenesis. AB - Interleukin-8 (IL-8, or CXCL8), which is a chemokine with a defining CXC amino acid motif that was initially characterized for its leukocyte chemotactic activity, is now known to possess tumorigenic and proangiogenic properties as well. In human gliomas, IL-8 is expressed and secreted at high levels both in vitro and in vivo, and recent experiments suggest it is critical to glial tumor neovascularity and progression. Levels of IL-8 correlate with histologic grade in glial neoplasms, and the most malignant form, glioblastoma, shows the highest expression in pseudopalisading cells around necrosis, suggesting that hypoxia/anoxia may stimulate expression. In addition to hypoxia/anoxia stimulation, increased IL-8 in gliomas occurs in response to Fas ligation, death receptor activation, cytosolic Ca(2+), TNF-alpha, IL-1, and other cytokines and various cellular stresses. The IL-8 promoter contains binding sites for the transcription factors NF-kappaB, AP-1, and C-EBP/NF-IL-6, among others. AP-1 has been shown to mediate IL-8 upregulation by anoxia in gliomas. The potential tumor suppressor ING4 was recently shown to be a critical regulator of NF-kappaB mediated IL-8 transcription and subsequent angiogenesis in gliomas. The IL-8 receptors that could contribute to IL-8-mediated tumorigenic and angiogenic responses include CXCR1 and CXCR2, both of which are G-protein coupled, and the Duffy antigen receptor for cytokines, which has no defined intracellular signaling capabilities. The proangiogenic activity of IL-8 occurs predominantly following binding to CXCR2, but CXCR1 appears to contribute as well through independent, small-GTPase activity. A precise definition of the mechanisms by which IL-8 exerts its proangiogenic functions requires further study for the development of effective IL-8-targeted therapies. PMID- 15831233 TI - Functional comparison of long and short splice forms of RPTPbeta: implications for glioblastoma treatment. AB - The receptor protein tyrosine phosphatase beta (RPTPbeta/PTPzeta) is overexpressed in glioblastoma tumors and plays a functional role in tumor cell migration and adhesion. Glioblastomas express at least three splice variants of RPTPbeta, including long and short receptor forms and a secreted chondroitin sulfate proteoglycan called phosphacan. Here we explore the differences in the expression pattern and function of long RPTPbeta and short RPTPbeta. The short form of RPTPbeta lacks exon 12, which encodes 860 amino acids located in the extracellular domain. Until now, functional differences between long and short RPTPbeta have been difficult to elucidate. In this study, antibodies specific to the splice junction, unique to short RPTPbeta, allowed for the discrimination of the two receptors. A study of normal brain tissue and graded astrocytomas indicates that long and short RPTPbeta forms have an overlapping expression pattern. In order to study functional differences between long and short RPTPbeta, we created stable U87 glioblastoma cells that expressed these receptors. U87 stable cell lines overexpressing long or short RPTPbeta migrate faster and adhere more robustly than parental U87 cells. The two forms differ in that long-RPTPbeta-overexpressing cells migrate and adhere better than short RPTPbeta-overexpressing cells. A study of the extracellular domain of short RPTPbeta indicates that it retains much of the functional capacity of phosphacan. Indeed, the action of recombinant, short-RPTPbeta extracellular domain protein is similar to that of phosphacan as a repulsive substrate for glioblastoma cells. Comparison of the signaling capacity of long RPTPbeta to that of short RPTPbeta reveals very similar abilities to activate transcription pathways. Moreover, transient transfection with either long or short RPTPbeta activates NF-kappaB reporter gene transcription. Because of their tumor-restricted and largely overlapping expression patterns in glioblastoma, both RPTPbeta splice forms are potential therapeutic targets. The involvement of long and short RPTPbeta in glioma tumor cell biology also contributes to the value of RPTPbeta as a cancer target. PMID- 15831234 TI - Patient tumor EGFR and PDGFRA gene amplifications retained in an invasive intracranial xenograft model of glioblastoma multiforme. AB - We have previously described a panel of serially transplantable glioblastoma multiforme xenograft lines established by direct subcutaneous injection of patient tumor tissue in the flanks of nude mice. Here we report the characterization of four of these lines with respect to their histopathologic, genetic, and growth properties following heterotopic-to-orthotopic (flank-to intracranial) transfer. Cells from short-term cultures, established from excised flank xenografts, were harvested and injected into the brains of nude mice (10(6) cells per injection). The intracranial tumors generated from these injections were all highly mitotic as well as highly invasive, but they lacked necrotic features in most instances and failed to show endothelial cell proliferation in all instances. For mice receiving injections from a common explant culture, tumor intracranial growth rate was consistent, as indicated by relatively narrow ranges in survival time. In contrast to the loss of epidermal growth factor receptor gene (EGFR) amplification in cell culture, high-level amplification and overexpression of EGFR were retained in intracranial tumors established from two EGFR-amplified flank tumors. A third intracranial tumor retained patient tumor amplification and high-level expression of platelet-derived growth factor receptor alpha gene. Because the heterotopic-to-orthotopic transfer and propagation of glioblastoma multiforme preserves the receptor tyrosine kinase (RTK) gene amplification of patient tumors, this approach should facilitate investigations for determining the extent to which RTK amplification status influences tumor response to RTK-directed therapies. The fact that such studies were carried out by using an invasive tumor model in an anatomically appropriate context should ensure a rigorous preclinical assessment of agent efficacy. PMID- 15831235 TI - Oral sodium phenylbutyrate in patients with recurrent malignant gliomas: a dose escalation and pharmacologic study. AB - We determined the maximum tolerated dose (MTD), toxicity profile, pharmacokinetic parameters, and preliminary efficacy data of oral sodium phenylbutyrate (PB) in patients with recurrent malignant gliomas. Twenty-three patients with supratentorial recurrent malignant gliomas were enrolled on this dose escalation trial. Four dose levels of PB were studied: 9, 18, 27, and 36 g/day. Data were collected to assess toxicity, response, survival, and pharmacokinetics. All PB doses of 9, 18, and 27 g/day were well tolerated. At 36 g/day, two of four patients developed dose-limiting grade 3 fatigue and somnolence. At the MTD of 27 g/day, one of seven patients developed reversible grade 3 somnolence. Median survival from time of study entry was 5.4 months. One patient had a complete response for five years, and no partial responses were noted, which yielded an overall response rate of 5%. Plasma concentrations of 706, 818, 1225, and 1605 muM were achieved with doses of 9, 18, 27, and 36 g/day, respectively. The mean value for PB clearance in this patient population was 22 liters/h, which is significantly higher than the 16 liters/h reported in patients with other malignancies who were not receiving P450 enzyme-inducing anticonvulsant drugs (P = 0.038). This study defines the MTD and recommended phase 2 dose of PB at 27 g/day for heavily pretreated patients with recurrent gliomas. The pharmacology of PB appears to be affected by concomitant administration of P450-inducing anticonvulsants. PMID- 15831232 TI - Hypoxia and the hypoxia-inducible-factor pathway in glioma growth and angiogenesis. AB - Glioblastomas, like other solid tumors, have extensive areas of hypoxia and necrosis. The importance of hypoxia in driving tumor growth is receiving increased attention. Hypoxia-inducible factor 1 (HIF-1) is one of the master regulators that orchestrate the cellular responses to hypoxia. It is a heterodimeric transcription factor composed of alpha and beta subunits. The alpha subunit is stable in hypoxic conditions but is rapidly degraded in normoxia. The function of HIF-1 is also modulated by several molecular mechanisms that regulate its synthesis, degradation, and transcriptional activity. Upon stabilization or activation, HIF-1 translocates to the nucleus and induces transcription of its downstream target genes. Most important to gliomagenesis, HIF-1 is a potent activator of angiogenesis and invasion through its upregulation of target genes critical for these functions. Activation of the HIF-1 pathway is a common feature of gliomas and may explain the intense vascular hyperplasia often seen in glioblastoma multiforme. Activation of HIF results in the activation of vascular endothelial growth factors, vascular endothelial growth factor receptors, matrix metalloproteinases, plasminogen activator inhibitor, transforming growth factors alpha and beta, angiopoietin and Tie receptors, endothelin-1, inducible nitric oxide synthase, adrenomedullin, and erythropoietin, which all affect glioma angiogenesis. In conclusion, HIF is a critical regulatory factor in the tumor microenvironment because of its central role in promoting proangiogenic and invasive properties. While HIF activation strongly promotes angiogenesis, the emerging vasculature is often abnormal, leading to a vicious cycle that causes further hypoxia and HIF upregulation. PMID- 15831236 TI - Phase 2 study of T138067-sodium in patients with malignant glioma: Trial of the National Cancer Institute of Canada Clinical Trials Group. AB - We studied the activity of T138067-sodium in patients with malignant gliomas. T138067-sodium is a unique new chemotherapy agent that inhibits microtubule formation by binding irreversibly and specifically to beta(1), beta(2)and beta(4) isotypes of 3-tubulin, causing cell arrest at G(2)/M and inducing apoptosis. Patients with recurrent anaplastic astrocytoma or glioblastoma multiforme were treated intravenously with 330 mg/m(2) of T138067-sodium weekly. Treatment was continued until the patient experienced either unacceptable toxicity or progressive disease. Patients had to have histologically proven glioma, have bidimensionally measurable disease at least 1 cm x 1 cm, and have received no more than one prior adjuvant chemotherapy. No chemotherapy or radiotherapy for recurrent disease was permitted. Nineteen patients entered the trial. One patient was found to be ineligible. There were two patients with anaplastic astrocytoma and 16 with glioblastoma multiforme. Only two patients had received prior adjuvant chemotherapy. The first seven patients had full pharmacokinetic sampling. No dose-limiting toxicity was seen, and pharmacokinetic results were consistent with those from nonglioma patients. The most common drug-related effects were fatigue (33%), nausea (28%), neutropenia (28%), and anorexia (17%). No patients stopped the study because of toxicity. No responses were seen in the 15 eligible patients who completed at least one cycle. Three patients had stable disease with a median duration of 2.6 months. Our results suggest that given in this dose and schedule T138067-sodium does not have activity in this population of anaplastic astrocytoma and glioblastoma multiforme. PMID- 15831237 TI - Cost of temozolomide therapy and global care for recurrent malignant gliomas followed until death. AB - Effectiveness and costs of care and treatment of recurrent malignant gliomas are largely unknown. In this study, 49 patients (32 males, 17 females; mean age, 49; age range, 23-79) were treated with temozolomide (TMZ) for recurrent or progressive malignant gliomas after standard radiation therapy. Cost assessment (payer's perspective) singled out treatment for first recurrence and all costs of care until death. We computed personnel costs as wages; drugs, imaging, and laboratory tests as prices; and hospitalizations as day rates. Patients were administered a median of five TMZ cycles at recurrence. Drug acquisition costs amounted to euro 2206 per cycle (76% of total costs). Seven patients showed no second recurrence (two are still alive), 16 received no further chemotherapy and died after 3.9 months, and 26 received second-line chemotherapy. After the second progression, median survival was 4.0 months (95% confidence interval, 1.8-6.1). Overall monthly costs of care varied between euro 2450 and euro 3242 among the different groups, and median cost-effectiveness and cost utility ranged from euro 28,817 to euro 38,450 and from euro 41,167 to euro 53,369 per life of year and per quality-adjusted life-year gained, respectively. We conclude that despite high TMZ drug acquisition costs, care of recurrent malignant gliomas is comparable to other accepted therapies. PMID- 15831238 TI - Consensus conference on cancer registration of brain and central nervous system tumors. AB - The passage of Public Law 107-260, the Benign Brain Tumor Cancer Registries Amendment Act, in October 2002 has made the collection of all primary brain tumors a reality. However, at the first Consensus Conference on Brain Tumor Definition for Registration in 2002, and during the development of training materials for benign brain tumor collection, several issues were identified that were tabled for future discussion. These and other issues were addressed at the subsequent 2003 Consensus Conference on Cancer Registration of Brain and Central Nervous System Tumors, at which the Central Brain Tumor Registry of the United States facilitated a discussion among epidemiologists, neurosurgeons, and neuropathologists. Multidisciplinary consensus was reached on four points, for which the following recommendations were made: (1) amend the histology coding scheme for cysts and tumor-like lesions that currently have a code in the third edition of the International Classification of Disease for Oncology (ICDO), (2) collect data on all instances of specific cysts and tumor-like lesions that are located in brain and other CNS sites but currently lack ICDO codes, (3) establish a new ICDO topography site for skull base tumors for the brain and CNS, and (4) collect data on genetic syndromes in patients diagnosed with brain or CNS tumors. We view this conference as part of a continuing process. Because classification of primary intracranial and other CNS tumors is dynamic, and the registration and coding of these tumors will need to be periodically reviewed. PMID- 15831242 TI - Degradation and synthesis kinetics of quorum-sensing autoinducer in Pseudomonas aeruginosa cultivation. AB - The quorum-sensing (las and rhl) systems play critical roles in the pathogenicity of Pseudomonas aeruginosa and its synthesis of the important biosurfactants, rhamnolipids. In this work, P. aeruginosa PAO1 and its rhlI and rhlR null mutants were used to study the degradation and synthesis kinetics of the rhl system's autoinducer PAI2 (N-butanoyl-homoserine lactone). The two mutants, lacking the ability of synthesizing PAI2 or RhlR protein, produced insignificant amounts of rhamnolipids while having similar growth profiles as the wild-type culture. The regulatory RhlR:PAI2 complex is thus essential to rhamnolipid synthesis. In batch culture of the wild-type PAO1, the autoinducer PAI2 concentration increased along cell growth, especially during the transition from exponential-growth phase to stationary phase, and began to decrease after entering the stationary phase. The decrease in the stationary phase resulted from a faster PAI2 degradation than its synthesis. The degradation kinetics was studied using PAI2-containing supernatants (from centrifuged broth of wild-type culture) with and without the rhlI(-) mutant cells incapable of PAI2 synthesis. Being insignificant in the cell free systems, PAI2 degradation was found predominantly cell-associated and could be described empirically by the first-order, exponential decay kinetics with the best-fit degradation constant (k(d)) of 0.195 h(-1). When similarly modeled with a first-order kinetics, PAI2 synthesis in stationary-phase wild-type culture was derived to have a synthesis constant (k(s)) of 0.189 h(-1). The PAI2 concentration in batch cultivation of the rhlR(-) mutant also showed an increase then-decrease profile. However, the maximum PAI2 concentration was about one third of that from the wild-type culture. The constitutive rate of PAI2 synthesis was therefore significantly lower than the rate attainable with active auto induction by RhlR-PAI2 complex. PMID- 15831243 TI - Heterologous expression of piglet odorant-binding protein in Pichia pastoris: a comparative structural and functional characterization with native forms. AB - This study targets to express the piglet odorant-binding protein (plOBP) and compare the engineered product to the corresponding native protein forms, i.e. plOBP and adult porcine OBP (pOBP). Using the natural signal peptide from the cDNA sequence, up to 40 mg l(-1) of secreted recombinant piglet OBP (rOBP) has been produced in a minimal culture medium. No significant difference in molecular mass between rOBP and native plOBP could be observed by mass spectrometry following or not trypsin digestion. rOBP and pOBP shared similar immunoreactivity towards polyclonal anti-pOBP antibodies, suggesting a proper processing and folding of the recombinant product. Both plOBP and rOBP displayed comparable binding properties towards fatty acids present in the putative maternal pheromone and a steroid, component of the boar sex pheromone. Furthermore, the rOBP product was found to bind to an olfactory receptor, for which pOBP binding was previously characterized. Taken together, these findings suggest that rOBP, produced in Pichia pastoris, exhibits structural and functional properties comparable to those of the native lipocalins from both young or adult animal. PMID- 15831244 TI - The CELO adenovirus Gam1 protein enhances transient and stable recombinant protein expression in Chinese hamster ovary cells. AB - The Gam1 protein of the avian CELO adenovirus activates transcription through inhibition of histone deacetylase 1 (HDAC1). We investigated the effect of Gam1 on both transient and stable transgene expression in Chinese hamster ovary (CHO) cells, one of the most commonly used mammalian hosts for the large-scale production of recombinant proteins. Transient expression of Gam1 increased reporter protein levels up to 4-fold in suspension cultures of CHO DG44 cells co transfected with a reporter gene and up to 20-fold in recombinant CHO DG44 derived cell lines. The highest levels of activation were observed when the transgene was under the control of the human cytomegalovirus (HCMV) immediate early promoter/enhancer. Increases in recombinant protein expression in the presence of Gam1 were not accompanied by an enhancement of cell growth or viability. We conclude that Gam1 may serve as a useful genetic tool for increasing recombinant protein expression in CHO DG44 cells. PMID- 15831245 TI - Enzymatic synthesis and anti-coagulant effect of salicin analogs by using the Leuconostoc mesenteroides glucansucrase acceptor reaction. AB - Glucansucrases from Leuconostoc mesenteroides catalyze the transfer of glucosyl units from sucrose to other carbohydrates by acceptor reaction. We modified salicyl alcohol, phenol and salicin by using various glucansucrases and with sucrose as a donor of glucosyl residues. Salicin, phenyl glucose, isosalicin, isomaltosyl salicyl alcohol, and a homologous series of oligosaccharides, connected to the acceptors and differing from one another by one or more glucose residues, were produced as major reaction products. By using salicin and salicyl alcohol as acceptors, B-1355C2 and B-1299CB-BF563 dextransucrases synthesized most widely diverse products, producing more than 12 and 9 different kinds of saccharides, respectively. With phenol, two acceptor products and oligosaccharides were synthesized by using the B-1299CB-BF563 dextransucrase. Salicyl derivatives, as acceptor products, showed higher anti-coagulation activity compared with that of salicin or salicyl alcohol that were used as acceptors. PMID- 15831246 TI - Purification and stabilization of ricin B from tobacco hairy root culture medium by aqueous two-phase extraction. AB - Ricin B (RTB), the non-toxic lectin subunit of ricin, is a promising mucosal adjuvant and carrier for use in humans. RTB fusion proteins have been expressed in tobacco hairy root cultures, but the secreted RTB component of these proteins was vulnerable to protease degradation in the medium. Moreover, castor bean purified RTB spiked into tobacco hairy root culture media showed significant degradation after 24 h and complete loss of product after 72 h. Aqueous two-phase extraction (ATPE) was tested for fast recovery of RTB not only to partially purify the protein but also to improve its stability. Two different polyethylene glycol (PEG)/salt/water systems including PEG/potassium phosphate and PEG/sodium sulfate, were studied. RTB was shown to be favorably recovered in PEG/sodium sulfate systems. Statistical analysis indicated that the ionic strength of the system and the sodium sulfate concentration were important in optimizing the partition coefficient of RTB. A selectivity of almost three could be achieved for RTB in optimized systems, and RTB partitioned in the PEG-rich phase exhibited extended stability. Therefore, ATPE was shown to be effective in initial recovery/purification and stabilization of RTB and may hold promise for other unstable secreted proteins from hairy root culture. PMID- 15831247 TI - Lipase-catalyzed synthesis of L-phenylalanyl-D-glucose. AB - Regioselective synthesis of L-phenylalanyl ester of D-glucose with unprotected L phenylalanine and D-glucose was carried out in organic solvents using lipases from Rhizomucor miehei (RML) and porcine pancreas (PPL). The reaction was investigated in terms of free unprotected L-phenylalanine, D-glucose, RML and PPL concentrations, buffer salts (pH and concentration), enzyme reusability and incubation period. Under the experimental conditions employed, both the enzymes exhibited good esterification potentialities, with RML exhibiting better conversions (maximum yield, 98%) than PPL (maximum yield, 75.6%). Reactions in the presence of buffer salts gave about 10% higher yields than those in their absence. Two-dimensional heteronuclear single quantum coherence transfer (HSQCT) NMR spectral analysis confirmed the formation of five diastereomeric esters: three different L-phenylalanyl-D-glucose monoesters (6-O: 24.1%, 3-O: 23.3% and 2 O: 19.2%) and two different diesters (2,6-di-O: 16.6% and 3,6-di-O: 16.8%) in an esterification yield of 92.4%. PMID- 15831248 TI - Effect of genetic background on glycosylation heterogeneity in human antithrombin produced in the mammary gland of transgenic goats. AB - Glycosylation is involved in the correct folding, targeting, bioactivity and clearance of therapeutic glycoproteins. With the development of transgenic animals as expression systems it is important to understand the impact of different genetic backgrounds and lactations on glycosylation. We have evaluated the glycosylation of recombinant antithrombin produced in several transgenic goat lines, from cloned animals and from different types of lactation including induced lactations. Our results show glycosylation patterns from the protein expressed in animals, derived from the same founder goat, are mostly comparable. Furthermore, the protein expressed in two cloned goats had highly consistent oligosaccharide profiles and similar carbohydrate composition. However, there were significantly different oligosaccharide profiles from the proteins derived from different founder goats. Artificial induction of lactation did not have significant effects on overall carbohydrate structures when compared to natural lactation. The only major difference was that recombinant antithrombin from induced lactations contained a slightly higher ratio of N-acetylneuraminic acid to N-glycolylneuraminic acid and less amount of oligosaccharides containing N glycolylneuraminic acid. The oligosaccharides from all animals were a mixture of high mannose-, hybrid- and complex-type oligosaccharides. Sialic acid was present as alpha-2,6-linkage and no alpha-1,3-linked galactose was observed. These results indicate that transgenic animals with closely related genetic backgrounds express recombinant protein with comparable glycosylation. PMID- 15831250 TI - Matrix assisted refolding of proteins by ion exchange chromatography. AB - Two different approaches of matrix assisted refolding have been evaluated and compared to conventional refolding by dilution. Bovine alpha-lactalbumin was used for the studies as model protein. It was adsorbed under denaturing conditions on an ion exchange matrix and refolding was completed on the column prior to elution or, depending on the buffer system, in the eluate. Agarose based chromatography matrices showed high capacities for the denatured alpha-lactalbumin. A positive effect on the yield of refolded protein by the matrix could be observed for Fractogel EMD DEAE and a negative for Toyopearl DEAE 650M, DEAE Sepharose FF and Q Sepharose FF. In the case of Fractogel EMD DEAE the ion exchange surface might act as a folding helper. This property may be caused by the grafted polymers. For Source 30Q only a marginal negative influence on the refolding kinetics was observed, thus the ion exchanger is only a mean for removal of chaotropic agents. Refolding on the column is characterized by a low yield but high productivity due to significant reduction of refolding time. PMID- 15831249 TI - Alkali and halide-resistant catalysis by the multipotent oxidase from Marinomonas mediterranea. AB - The incorporation of fungal laccases into novel applications has been delayed mainly due to their intrinsic sensitivity towards halides and alkaline conditions. In order to explore new sources of enzymes we evaluated the multipotent polyphenol oxidase PPO1 from the marine bacterium Marinomonas mediterranea. Here we report that, in contrast to its fungal counterparts, PPO1 remained functional above neutral pH presenting high specificity for phenolic compounds, in particular for methoxyl-substituted mono-phenols and catechols. These properties, in addition to its tolerance towards chloride (up to 1 M) and its elevated redox potential at neutral pH (0.9 V), suggest this enzyme may be an interesting candidate for specific applications such as the Amperometric determination of phenolic compounds and bio-fuel cells. PMID- 15831251 TI - Scalable encapsulation of hepatocytes by electrostatic spraying. AB - Encapsulating cells by polyelectrolyte complex coacervation can be accomplished at physiological temperature and buffer conditions. One of the oppositely charged polyelectrolytes in the microcapsule core can be collagen or any other natural extra-cellular matrices suitable for cellular support while the other polyelectrolyte forms the ultra-thin shell to ensure efficient mass transfer. These microcapsules with ultra-thin shell are difficult to produce in large quantities due to their fragility. In this study, electrostatic spraying technique was used to achieve a scalable production of one such type of microcapsules formed by complex coacervation between the cationic methylated collagen and anionic terpolymer of hydroxylethyl methacrylate, methyl methacrylate and methylacrylic acid (HEMA-MMA-MAA). It was found that the microcapsule sizes were dependent on several important operational parameters, such as the diameter of the spraying needle, the flow rate of the hepatocytes collagen mixture and the voltage of the electrical field. The microcapsules with diameters of 200-800 microm and a narrow size distribution (standard deviation of 5-28%) were successfully produced. The above parameters also influenced the hepatocyte viability and functions. With a practical encapsulation rate of up to 55 ml/h per orifice required in bio-artificial liver-assisted device applications, we have produced large quantities of microcapsules maintaining comparable cell viability (>87%), mechanical stability and bio-functions to the manually extruded microcapsules. PMID- 15831252 TI - Improved production of L-lysine by disruption of stationary phase-specific rmf gene in Escherichia coli. AB - Growth and rate, at which fermentation products are formed in cells, generally decreases during the stationary phase as a result of changes in gene expression. We focused on the rmf gene, which encodes the ribosome modulation factor protein, as a target for strain modification in order to improve the rate of L-lysine production in Escherichia coli. Increased expression of the rmf gene during the stationary phase was confirmed under various cultivation conditions using DNA macroarray analysis. Mutants with disrupted rmf were then generated from an L lysine-producing E. coli strain. The rates of L-lysine accumulation and production were significantly increased in disruptants that were cultivated with excess phosphate. By contrast, a higher biomass was generated in disruptants that were grown under limited phosphate conditions. These results demonstrate that disruption of the rmf gene significantly affects L-lysine production and growth in E. coli. PMID- 15831253 TI - Possibilities for controlling a PHB accumulation process using various analytical methods. AB - Poly-beta-hydroxybutyrate (PHB) and other polyesters can be produced by various species of bacteria. Of the possible carbon sources, methane could prove to be one of the most suitable substrates for the manufacture of PHB. The methanotrophic strain Methylocystis sp. GB 25 DSM 7674 was applied in order to accumulate PHB in a rapid, non-sterile process. Cultivation was performed in two stages: a continuous growth phase (dilution rate 0.17 h(-1)) and a PHB accumulation phase under deficiency conditions of an essential nutrient (e.g. phosphorus) in batch culture. The PHB content of the biomass was as high as 51%; efficiency was the highest during the first 5 h of the product formation process. The PHB produced is of very high quality, having a high molecular mass of up to 2.5 x 10(6) Da. In order to monitor and control the process, a rapid analysis method based upon turbidimetry in the visible range (438 nm) was applied. Moreover, the PHB content of the biomass was determined using an FT-IR spectroscopic method with ATR sampling and multivariate calibration. We achieved a value of 1.4% as the best standard error of cross validation. The nitrogen content of the PHB final product (a product quality parameter) was estimated by spectroscopic method in the visible range. PMID- 15831255 TI - The first metatarsal bone under loading conditions: a finite element analysis. AB - An individual-based, three-dimensional finite element model of the first metatarsal (MT I) bone was created with fine CT. The three-dimensional model of the bone was fixed proximally at the metatarsocuneiform joint and load was applied on the metatarsal head. Loading conditions were simulated, including muscular forces as described for a normal metatarsophalangeal (MTP) joint during three typical phases of gait as the combination of the load in the contact areas of the sesamoid bones and the base of the proximal phalanx. The resultant strain and stress distributions within the loaded MT I were calculated and visualized with the MTP in different positions. PMID- 15831256 TI - Osteotomies for hallux valgus correction. AB - A variety of metatarsal osteotomies has been described since the first report by Gernet in 1836. Many of these osteotomies were abandoned throughout the years. Because one procedure is not capable of correcting all types of hallux valgus deformities, an algorithm, as a guideline, is preferable. This article presents a choice of metatarsal osteotomies for correction of hallux deformities based on clinical and biomechanical research. PMID- 15831257 TI - First metatarsal osteotomy nonunion and malunion. AB - The purpose of this article is to review our experience with malunion and nonunion of proximal and distal first metatarsal osteotomies and to outline the treatment options when such complications occur. PMID- 15831258 TI - Management of soft tissue deficiency of the hallux: salvage in trauma, diabetes, and following surgical complications. AB - Loss of the hallux can lead to significant gait problems and cause difficult biomechanical problems with weight bearing. Amputation of the first ray should be avoided when possible. Soft tissue defect of the hallux can be particularly difficult to manage because of the absence of local muscle tissue for coverage. In younger patients, soft tissue loss usually is related to trauma; an understanding of the techniques that are available for coverage are important to maintain length and function of the first ray. Diabetic neuropathy and resultant ulceration of the hallux or metatarsophalangeal joint is another common cause of soft tissue deficiency that may lead to deep infection and potentially result in amputation. Certain systemic problems, such as inflammatory diseases or gout, can cause significant degeneration of the local tissues which also can be problematic. Wound dehiscence or loss of skin flaps that are created during surgery represent further difficulties that may be encountered. This article reviews the common causes of amputation of the hallux and the principles that are necessary for salvage of the digit. PMID- 15831259 TI - Surgical treatment of the hallux rigidus. AB - Numerous surgical procedures are available for the treatment of hallux rigidus. The choice of a particular one depends on the severity of the disease, patient activity level, and expectations about the surgery. Cheilectomy is recommended for early disease and may be associated with an osteotomy of the proximal phalanx. For active patients who have severe hallux rigidus, arthrodesis and biologic interposition arthroplasties have shown good results. Keller arthroplasty is reserved for low demand patients; prosthetic replacements are not recommended at this time. PMID- 15831260 TI - Joint replacement in the hallux metatarsophalangeal joint. AB - Joint replacement of the hallux metatarsophalangeal joint has not enjoyed the same success as hip and knee arthroplasties. Silastic implants have achieved a high patient satisfactory rate but have caused many complications, including silicone synovitis and lymph node inflammation. Metal and polyethylene hemiarthroplasties and total toe replacements seem to be more promising although results are preliminary. Problems with these implants seem to be related to soft tissue instability of the joint; patients who have hallux rigidus have more success than patients who have hallux valgus or rheumatoid arthritis. Severe complications can be treated with removal and synovectomy or arthrodesis, depending on the length and alignment of the foot, as well as the functional demands of the patients. It would be beneficial to have more data on these implants so that improvements can be made in design and patient selection. PMID- 15831261 TI - Avascular necrosis of the first metatarsal head. AB - Avascular necrosis of the first metatarsal head is uncommon. It is most often seen following a distal metatarsal osteotomy for hallux valgus. In this setting surgery has usually involved extensive periarticular dissection as well. Although many cases may be subclinical, in its most pronounced form it is a powerful cause of failure of bunion surgery. This article examines the underlying factor contributing to this problem as well as its long-term management. PMID- 15831262 TI - Disorders of the hallucal interphalangeal joint. AB - Most clinical presentations of the hallux concern the metatarsophalangeal joint; however, interphalangeal joint (IPJ) pathology also may be clinically significant. This article reviews conditions that commonly affect the hallucal IPJ and the appropriate treatment. PMID- 15831263 TI - The great toe proximal phalanx osteotomy: the final step of the bunionectomy. AB - As the final step of correction of hallux valgus deformity, the great toe proximal phalanx osteotomy is useful. It is popular in France and throughout Europe. Our purpose is to distinguish and to describe different locations and three types of osteotomies according to the required final correction of the hallux valgus. PMID- 15831264 TI - Hypermobility of the first tarsometatarsal joint. AB - This article discusses hypermobility of the first tarsometatarsal joint. PMID- 15831265 TI - Technical considerations in hallux metatarsalphalangeal arthrodesis. AB - Metatarsalphalangeal (MTP) arthrodesis remains the gold standard for arthritis and salvage of the painful first MTP joint. There is a variety of techniques, approaches, and armamentarium to consider, depending on the clinical presentation and host factors. This article reviews these presents the author's personal experience. PMID- 15831266 TI - Orthotic management for disorders of the hallux. AB - Most disease conditions that affect the hallux are amenable to conservative treatment. Most of the literature that addresses the treatment of hallux rigidus, bunions, sesamoid problems, and arthritis focus on surgical management, although operative indications stress the need for attempted conservative treatment of these processes. This article reviews the technical considerations that are required for successful orthotic management of disease of the first ray. Mechanical terminology and principles of management are reviewed and examples of orthotic prescriptions for specific disease entities are included to facilitate communication between the orthopedist and orthotist. PMID- 15831268 TI - Epidemiology of hepatocellular carcinoma. AB - Year 2000 estimates of the incidence of cancer indicate that primary liver cancer remains the fifth most common malignancy in men and the eighth in women. The number of new cases has been predicted as 564,000, corresponding to 398,000 in men and 166,000 in women. The geographic areas at highest risk are located in Eastern Asia, Middle Africa, and some countries of Western Africa. Changes in incidence among migrant populations underline the predominant role of environmental factors in the etiology of primary liver cancer. In high-risk countries, the early cases of primary liver cancer occur already at ages 20 and above, underlying the impact of viral exposures early in life. In countries at low risk, primary liver cancer is rare before the 50s, translating the impact of late exposures with moderate risks and long latency intervals. Sex ratios are typically between 2 and 4. The incidence of primary liver cancer is increasing in several developed countries including the United States, and the increase will likely continue for several decades. The trend has a dominant cohort effect related to exposures to hepatitis B and C viruses. The variability of primary liver cancer incidence is largely explained by the distribution and the natural history of the hepatitis B and C viruses. The attributable risk estimates for the combined effects of these infections account for well over 80% of liver cancer cases worldwide. Primary liver cancer is the first human cancer largely amenable to prevention using hepatitis B virus vaccines and screening of blood and blood products for hepatitis B and C viruses. PMID- 15831269 TI - Staging for liver cancer. AB - This article focuses on the development of prognostic scores, their use in the management of cancer patients, and how they are different from staging systems. It is underlined that the development of a prognostic score is a dynamic process. The score can be modified by incorporating new prognostic features provided that appropriate methodology is followed. The peculiarity of hepatocellular carcinoma patients are addressed, and each individual prognostic systems for hepatocellular carcinoma is presented and discussed. PMID- 15831270 TI - The use of decision analytic models to inform clinical decision making in the management of hepatocellular carcinoma. AB - Decision analysis helps evaluate competing strategies under conditions of uncertainty in a wide variety of clinical settings. Despite some limitations, decision trees and Markov models remain essential tools for medical decision analysts. These techniques allow comparison of competing management strategies in a quantitative fashion. Sensitivity analysis is an important feature of decision analytic models that identify important factors that affect the outcome of decisions under considerations. Judiciously used, decision analytic models allow a quantitative evaluation of existing data as they relate to strategies ranging from optimizing clinical management at the patient level to allocating health care resources at the societal level. PMID- 15831271 TI - Screening tests for hepatocellular carcinoma. AB - The incidence of hepatocellular carcinoma (HCC) is rising throughout the world. HCC meets the criteria for which a disease benefits from screening or surveillance: it is an important health problem; those with cirrhosis are the targets for surveillance; there is acceptable treatment if diagnosed early; surveillance using alpha-fetoprotein and ultrasound has been shown to be cost effective; surveillance is widely implemented by health care professionals and accepted by patients; standardized recall procedures exists; and the screening tests must achieve an acceptable level of accuracy in the population undergoing screening. The latter point is the main limitation of surveillance for HCC. In this review we will discuss the currently available tests for the surveillance of HCC. PMID- 15831272 TI - Imaging diagnosis of hepatocellular carcinoma with differentiation from other pathology. AB - Recent advances in liver imaging techniques and better understanding of imaging findings have facilitated the detection and characterization of hepatocellular nodules in a cirrhotic liver. It is important to recognize that various types of benign nodules and pseudolesions are identified on all imaging scans performed for the diagnosis of hepatocellular carcinoma. An accurate differentiation between them is critical for adequate management of cirrhotic patients. Unfortunately, any of the imaging tests and even percutaneous biopsy are not diagnostic for borderline lesions. Intimate collaboration of hepatologists, pathologists, surgeons, and radiologists with reasonable imaging and clinical criteria estimating the degree of malignancy is imperative. PMID- 15831273 TI - Liver biopsy in the diagnosis of hepatocellular carcinoma. AB - Dysplastic nodules are the precursor lesions of hepatocellular carcinoma (HCC). Accurate diagnosis of dysplastic nodules and well-differentiated HCC is difficult with biopsy samples. Lesions often have regional variation of severity. Invasion of stroma, although a useful criterion of carcinoma, is seldom found on needle biopsies. Many criteria of neoplasia, such as widened plates and mitotic activity, are also found in reactive states. Thus, clinical history needs to be taken into consideration. No single criterion is sufficient for diagnosis of HCC. The best criteria for differentiation from dysplastic nodules on needle biopsies are (1) liver cell plates more than two cells in width or atypical plate structure, (2) high N/C ratio, and (3) nuclear atypia. The Laennec Classification of Hepatic Neoplasia may assist the standardization of these criteria. PMID- 15831274 TI - Chemoembolization and bland embolization: a critical appraisal. AB - Hepatocellular carcinoma is a major cause of cancer-related mortality worldwide, and most patients are not candidates for potentially curative treatment. Bland and chemoembolization are palliative options for hepatocellular carcinoma (HCC) that have been evaluated in controlled trials. Chemoembolization and perhaps bland embolization used as primary treatment for HCC in selected patients are effective at prolonging survival. The role of these therapies before surgical resection, liver transplantation, or in combination with local ablative therapy is controversial and yet unproven. PMID- 15831275 TI - A critical appraisal of the literature on local ablative therapies for hepatocellular carcinoma. AB - Image-guided percutaneous ablation is currently accepted as the best therapeutic option for nonsurgical patients with early-stage hepatocellular carcinoma. Ethanol injection is the seminal technique for local tumor treatment, and may achieve 50% 5-year survival in selected Child A patients. Radiofrequency ablation constitutes the most assessed alternative technique. On the basis of the identified evidence, radiofrequency ablation seems to reach higher recurrence free survival rates compared with ethanol injection. Further randomized trials are needed to establish the clinical efficacy of radiofrequency ablation with respect to other percutaneous treatments and to devise an unbiased therapeutic strategy. PMID- 15831276 TI - Liver transplantation for hepatocellular carcinoma: is expansion of criteria justified? AB - Hepatocellular carcinoma (HCC) is a major health problem worldwide, involving more than a half million new cases yearly, with an age-adjusted incidence of 5.5 to 14.9 per 10(5) population. In some areas of Asia and the Middle East, HCC ranks as the first cause of death due to cancer. The incidence of HCC is increasing in Europe and the United States, and it is currently the leading cause of death among cirrhotic patients. It is estimated that by the year 2010, the number of patients with HCC awaiting liver transplant in the United States will outnumber the supply of cadaver organs. PMID- 15831277 TI - Prevention of hepatocellular carcinoma. AB - The accuracy and the reliability of well-recognized clinical, virologic, histologic, and molecular risk factors for hepatocellular carcinoma are still insufficient; thus, accurate risk prediction of developing cancer in individual patients remains an elusive goal. PMID- 15831278 TI - Effect of serum and cumulus cell expansion on marker gene transcripts in bovine cumulus-oocyte complexes during maturation in vitro. AB - OBJECTIVE: To determine the distribution of transcripts encoding the FSH receptor (FSHr), LH receptor (LHr), connexin 43 (Cx43), cyclooxygenase-2 (COX-2), and prostaglandin E(2) receptors 2 and 3 (EP2 and EP3) within bovine cumulus-oocyte complexes (COCs) and denuded oocytes and investigate the influence of gonadotropins, serum, and cumulus cell expansion on the abundance of transcripts encoding these genes. DESIGN: Prospective controlled animal study. SETTING: University research laboratory. PATIENT(S): Animal models for human studies. INTERVENTION(S): Cumulus-oocyte complexes were treated in culture with serum and gonadotropin-supplemented media to examine the effects to mRNA transcript levels. MAIN OUTCOME MEASURE(S): Variation in mRNA transcript levels. RESULT(S): Luteinizing hormone receptor, FSHr, and EP3 mRNAs were detected in intact COCs and not in cumulus cell-denuded oocytes, whereas Cx43, COX-2, and EP2 mRNAs were found in both COCs and oocytes. The relative abundance of marker gene mRNAs did not vary in media containing no additives or FSH alone, independent of whether the media induced cumulus cell expansion. However, the presence of serum in maturation media significantly decreased expression of all mRNAs except LHr. CONCLUSION(S): The relative abundance of COC mRNAs is altered by serum in the maturation medium, which may signify long-term consequences for embryonic development. PMID- 15831279 TI - Reduction of estrogen production by interleukin-6 in a human granulosa tumor cell line may have implications for endometriosis-associated infertility. AB - OBJECTIVE: To examine the effect of interleukin-6 (IL-6) on estrogen production and aromatase activity using a human granulosa tumor cell line (KGN cells). The involvement of the mitogen-activated protein kinase (MAPK) cascade in the inhibitory effects of IL-6 on estrogen production was also evaluated. DESIGN: Molecular and biological studies of KGN cells. SETTING: Department of Obstetrics and Gynecology, Tottori University Hospital, Yonago, Japan. MAIN OUTCOME MEASURE(S): Gene expression of IL-6 and the IL-6 receptor was analyzed by reverse transcription-polymerase chain reaction and Southern blot analysis. KGN cells were cultured for 48 hours with IL-6 (0.1-10 ng/mL) or IL-6 (10 ng/mL) plus a mitogen activated protein kinase-extracellular signal regulated kinase kinase 1/2 (MEK1/2) inhibitor U0126 (10 microM). Estradiol concentration in the culture supernatants was measured by means of enzyme immunoassay, [1beta-(3)H] androstenedione was added to the cell lysate supernatant, and aromatase activity was determined by measuring the amount of [(3)H] H(2)O released upon the conversion of [1beta-(3)H] androstenedione to estrone. To examine the activation of intracellular signal transduction molecules induced by IL-6, the phosphorylation of Stat3, p38 MAPK, and extracellular signal-regulated kinase 1/2 (ERK1/2) was examined by Western blotting. RESULT(S): Gene expression of IL-6 and its receptor was detected in KGN cells. Estradiol secretion was significantly inhibited by adding IL-6, which also suppressed aromatase activity to 50% of the control. In addition, pretreatment with U0126 restored the IL-6-induced suppression of aromatase activity. IL-6 markedly enhanced the phosphorylation of ERK1/2, but not Stat3 and p38 MAPK. U0126 markedly reduced the level of the IL-6 induced phosphorylation of ERK1/2. CONCLUSION(S): These findings demonstrate that IL-6 may reduce estrogen production via the MAPK signal pathway in human granulosa cells. The results may support the notion that IL-6 is related to impaired estrogen biosynthesis in patients with endometriosis. PMID- 15831280 TI - Nicotine induces apoptosis in TM3 mouse Leydig cells. AB - OBJECTIVE: To investigate whether nicotine-induced testicular toxicity involves the induction of apoptosis in Leydig cells. DESIGN: Cell study using cells of the TM3 cell line derived from mouse Leydig cells. SETTING: Academic research laboratory. INTERVENTION(S): Morphological and biochemical analyses for the detection of apoptosis. MAIN OUTCOME MEASURE(S): The effect of nicotine on the occurrence of apoptosis was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay, 4,6-diamidino-2-phenylindole staining, terminal deoxynuclotidyl transferase-mediated dUTP nick end labeling assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction, caspase-3 enzyme assay, and Western blot analysis. Nicotine treatment exhibits several features of apoptosis in mouse Leydig cells. RESULT(S): TM3 cells treated with nicotine exhibit several features of apoptosis. It was also shown that nicotine increases the mRNA level of bax and decreases that of bcl-2. In addition, nicotine enhanced the expression of the activated form of caspase-3 and caspase-3 enzyme activity. CONCLUSION(S): Nicotine appears to activate specific intracellular death-related pathways, probably by bax-dependent activation of caspase-3, inducing apoptosis in Leydig cells. Thus, nicotine-induced apoptosis of Leydig cells might be one of the important mechanisms behind nicotine-related urogenital disorders in men. PMID- 15831281 TI - Expression levels of the inhibitor of apoptosis survivin in testes of patients with normal spermatogenesis and spermatogenic failure. AB - OBJECTIVE: To determine the expression of the inhibitor of apoptosis survivin in men with and without spermatogenic failure. DESIGN: Prospective case study. SETTING: Two university-based infertility clinics. PATIENT(S): Forty-nine infertile men presenting with azoospermia. INTERVENTION(S): Testicular biopsies for histopathological assessment and analyses of survivin expression levels by real-time reverse transcription-polymerase chain reaction. Survivin levels were normalized to expression of the housekeeping porphobilinogen deaminase gene. MAIN OUTCOME MEASURE(S): Correlation of the histological findings with normalized survivin expression levels. RESULTS(S): Testicular survivin mRNA expression was highest in normal spermatogenesis (n = 14). Decreased expression was observed in patients with spermatogenesis disorders. The expression level correlated with the degree of spermatogenic failure. While it was reduced in postmeiotic maturation arrest (n = 11), a lack of expression was seen in most specimens (10 of 12) with premeiotic maturation arrest and in all of those with Sertoli cell-only syndrome (n = 12). CONCLUSION(S): These data indicate that survivin is expressed in human germ cells and might be involved in apoptosis control during spermatogenesis. Decreased survivin expression in spermatogenic disorders may contribute to the accelerated germ cell apoptosis observed in male idiopathic infertility. PMID- 15831282 TI - Caspase activation in human spermatozoa in response to physiological and pathological stimuli. AB - OBJECTIVE: To investigate caspase activation in response to a variety of pathological and physiological stimuli in light of the fact that current research offers no clear consensus about caspase activation pathways in spermatozoa. DESIGN: A prospective, controlled study. SETTING: Male infertility clinic, Glickman Urological Institute, Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S): Fifteen healthy volunteers. INTERVENTION(S): Spermatozoa were exposed to [1] Fibroblast-associated (Fas) death receptor activation, [2] mitochondrial apoptosis induction using betulinic acid, [3] oxidative stress, and [4] prolonged incubation up to 3 hours without any external stimuli. MAIN OUTCOME MEASURE(S): Active caspases-1, -3, -8, and -9 were examined in human spermatozoa by flow cytometry using carboxyfluorescein derivatives. RESULT(S): Inducing Fas antibody did not result in any caspase activation. Conversely, betulinic acid significantly triggered caspase-9 and -3 activation. The application of oxidative stress and prolonged incubation (3 hours) failed to result in caspase activation. CONCLUSION(S): These results suggest that Fas has no functional relevance in mediating caspase activation in human ejaculated spermatozoa. Although spermatozoal mitochondria are highly susceptible to specific agonists of apoptosis such as betulinic acid via caspase activation, oxidative stress-induced apoptosis appears to be caspase independent. PMID- 15831283 TI - Efficacy of three adhesion-preventing agents in reducing severe peritoneal trauma induced by bipolar coagulation in a laparoscopic rat model. AB - OBJECTIVE: To compare the antiadhesion efficacy of three agents (4% icodextrin, ferric hyaluronate gel, and Ringer's lactate) in severe peritoneal trauma caused by bipolar coagulation in a laparoscopic rat model. DESIGN: Prospective, randomized, blinded experimental study. SETTING: International Laparoscopic Surgery Centre, Clermont-Ferrand, France. ANIMAL(S): Female Sprague-Dawley rats. INTERVENTIONS: Animals were prospectively randomized to receive one of the three agents or to be controls. Coagulation of the parietal peritoneum was performed using a bipolar 40-W current followed by administration of an antiadhesion agent. Postoperative adhesion assessment was carried out on day 7 in 42 rats. MAIN OUTCOME MEASURE(S): Adhesions were scored according to their extent and severity. RESULT(S): Adhesions occurred in all rats, however, adhesion and vascular adhesion rate scores were significantly higher in controls compared with treated groups. Adhesions were more likely to be filmy and easily separable in the 4% icodextrin group compared with the Ringer's lactate solution group. CONCLUSION(S): Adhesion scores were decreased by the use of antiadherent agents. However, no rats were found to be free of adhesions after severe peritoneal trauma induced by 40-W bipolar coagulation of the peritoneum. PMID- 15831284 TI - Prolactin inhibits oocyte release after gonadotropin stimulation in the rat: putative mechanism involving ovarian production of beta-endorphin and prostaglandin. AB - OBJECTIVE: To evaluate whether prolactin (PRL) is able to inhibit ovulation induced with exogenous gonadotropins in the rat and whether this effect could be mediated by the ovarian production of beta-endorphin, prostaglandin, and nitric oxide (NO). DESIGN: Controlled in vivo and in vitro experiments. SETTING: Academic research laboratories. ANIMAL(S): Immature female rats undergoing ovulation induction with equine gonadotropins and hCG. INTERVENTION(S): Prolactin (100 or 200 microg), PRL + the opioid antagonist naloxone (200 microg each), or placebo were injected SC 4 hours after hCG administration for ovulation induction. In the in vitro experiments, isolated preovulatory ovaries were incubated with or without PRL in a final concentration of 100 or 200 ng/mL. MAIN OUTCOME MEASURES(S): Number of oocytes ovulated in vivo, ovarian beta-endorphin, PGE(2) and NO(2)(-)/NO(3)(-) release, and NO synthase activity in vitro. RESULT(S): Prolactin reduced significantly the number of oocytes ovulated at the doses of 100 and 200 microg, and this effect was partially reversed by naloxone administration together with 200 mug PRL. PRL also induced a twofold increase in the ovarian release of beta-endorphin and a threefold decrease in the ovarian production of PGE(2). Ovarian NO synthase activity and the concentrations of NO(2)(-)/NO(3)(-) in the incubation medium were not modified by PRL. CONCLUSION(S): Prolactin is able to reduce the number of oocytes released and modulate ovarian beta-endorphin and PGE(2) release, which may account for its peripheral anovulatory effects. This local effect of PRL could interfere in the process of ovulation induction by exogenous gonadotropins. PMID- 15831285 TI - Effective treatment of experimental human endometrial cancers with targeted cytotoxic luteinizing hormone-releasing hormone analogues AN-152 and AN-207. AB - OBJECTIVE: To treat experimental human endometrial cancers based on targeted chemotherapy with the cytotoxic luteinizing hormone-releasing hormone (LHRH) analogues AN-152 and AN-207. DESIGN: Experimental study using athymic nude mice bearing xenografts of HEC-1A and RL-95-2 human endometrial cancers to assess the efficacy and toxicity of AN-152 and AN-207. The expression of LHRH receptors in HEC-1A and RL-95-2 cancers was determined by reverse transcription-polymerase chain reaction, Western blot analysis, and radioligand binding assays. SETTING: Experimental laboratory research. ANIMAL(S): Female athymic nude mice (Ncr, nu/nu). INTERVENTION(S): Animals were treated with IV injections of the cytotoxic LHRH analogues AN-152 and AN-207 and their respective cytotoxic radicals doxorubicin (DOX) and AN-201 (2-pyrrolinodoxorubicin) on a control vehicle solution. MAIN OUTCOME MEASURE(S): Tumor volume, final tumor weight, tumor doubling time, body weight, white blood cell count, and LHRH receptor expression. RESULT(S): AN-152 significantly inhibited the growth of HEC-1A tumors. AN-207 also significantly suppressed the proliferation in vivo of HEC-1A and RL-95-2 cancers. The cytotoxic radicals DOX and AN-201 had no effect. Furthermore, mRNA for LHRH receptors, LHRH receptor protein, and high-affinity binding sites for LHRH were demonstrated on tumors. CONCLUSION(S): Targeted chemotherapy with AN 152 and AN-207 strongly inhibits the growth of human endometrial cancers, which express LHRH receptors, and could provide a new treatment modality for women with advanced endometrial carcinoma. PMID- 15831286 TI - Allelic loss studies do not provide evidence for the "endometriosis-as-tumor" theory. AB - OBJECTIVE: To identify consistent genetic changes in endometriosis samples to determine whether endometriosis lesions are true neoplasms. DESIGN: We analyzed ovarian endometriosis lesions for loss of heterozygosity (LOH) at 12 loci of potential importance (D9S1870, D9S265, D9S270, D9S161, D11S29, D1S199, D8S261, APOA2, PTCH, TP53, D10S541, and D10S1765), including some at which genetic changes were previously reported in endometriosis. SETTING: Molecular biology laboratory in a university hospital department. PATIENT(S): Seventeen women with ovarian endometriosis. INTERVENTION(S): Laser capture microdissection to separate the endometriotic epithelium, the adjacent endometriotic stroma, and surrounding normal ovarian stromal tissue, followed by DNA extraction and polymerase chain reaction amplification of polymorphic microsatellite markers. MAIN OUTCOME MEASURE(S): Fluorescence-based quantitation for the LOH analysis. RESULT(S): We identified LOH in only one lesion at one locus (D8S261). CONCLUSION(S): Our data do not support the hypothesis that ovarian endometriosis is a true neoplasm. PMID- 15831287 TI - Increases in phosphorylation of SAPK/JNK and p38MAPK correlate negatively with mouse embryo development after culture in different media. AB - OBJECTIVE: To test whether signal transduction proteins that mediate stress may be used to detect responses of embryos to different media in a prospective randomized study. DESIGN: Controlled laboratory study. SETTING: None. PATIENT(S): None. INTERVENTION(S): Mouse embryos isolated at E3.5 (3.5 days after fertilization) or E1.5 were cultured in different media for 24 hours or 72 hours, respectively. Expression of p38 mitogen activated protein kinases (MAPKs) and stress-activated protein kinase/Jun kinase (SAPK/JNK) phosphoproteins in the mouse embryo and their correlation with preimplantation development were studied. MAIN OUTCOME MEASURE(S): [1] In E3.5 embryos, SAPK/JNK and p38MAPK are phosphorylated at different levels in different media after 24 hours, with Ham's F10+BSA and M-16 having the highest intensity of both SAPK/JNK and p38MAPK phosphorylation and Quinn's cleavage medium and potassium simplex optimized medium supplemented with amino acids (KSOM+AA) the lowest intensity. [2] The stress-induced increase in phosphorylation of SAPK/JNK and p38MAPK appears to be post-translational in embryos. [3] The intensity of SAPK/JNK phosphorylation measured at E1.5+72 hours culture is inversely correlated with 4-cell/compaction rate, morula formation rate, blastocyst formation rate, and hatching rate. RESULT(S): SAPK/JNK and p38MAPK phosphoprotein levels, but not all forms of protein, are affected during culture of preimplantation embryos in seven different media. During culture, the rate of progress to four developmental events was assayed and each rate was inversely proportional to the level of SAPK/JNK phosphorylation measured by immunocytochemical means or Western blot analysis at the end of culture. CONCLUSION(S): Culture stresses embryos; different media exert different levels of stress on the embryos. There is a negative correlation between the amount of stress and the development rate. Taken together, the data suggest that SAPK/JNK phosphorylation may constitute a measure of homeostatic response to negative stimuli of media. PMID- 15831288 TI - Basal release of urokinase plasminogen activator, plasminogen activator inhibitor 1, and soluble plasminogen activator receptor from separated and cultured endometriotic and endometrial stromal and epithelial cells. AB - OBJECTIVE: To investigate whether separated and cultured endometriotic and endometrial stromal and epithelial cells release urokinase plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), and soluble plasminogen activator receptor (suPAR) antigens in vitro. DESIGN: In vitro study. SETTING: University hospital clinic. PATIENT(S): Regularly menstruating women with and without endometriosis. INTERVENTION(S): Tissue samples were collected at surgery performed for clinical reasons. MAIN OUTCOME MEASURE(S): The antigen concentrations of uPA, PAI-1, and suPAR in culture medium were assayed by enzyme linked immunosorbent assay. RESULT(S): Both stromal and epithelial cells from endometriotic and endometrial tissue released the three types of antigens, but the release of PAI-1 was significantly higher from stromal cells in the three types of tissue than from epithelial cells. Furthermore, the release of PAI-1 was significantly higher from endometriotic cells than from endometrial stromal cells. CONCLUSION(S): This study has demonstrated the basic capacity of separated epithelial and stromal cells from all three types of tissue to release uPA, PAI 1, and suPAR without any paracrine influence, as in vivo. The higher release of PAI-1 from endometriotic stromal cells might have importance for the invasive growth. PMID- 15831289 TI - Effect of dehydroepiandrosterone on central and peripheral levels of allopregnanolone and beta-endorphin. AB - OBJECTIVE: To evaluate the effects of dehydroepiandrosterone (DHEA) oral administration on neuroendocrine function by investigating the modulation exerted by DHEA administration on allopregnanolone and beta-endorphin (beta-EP) central and peripheral levels in ovariectomized rats. DESIGN: Prospective study. SETTING(S): Experimental research environment. ANIMAL(S): Female Wistar rats (n = 48). INTERVENTION(S): Forty rats were ovariectomized and received an oral treatment with either placebo or 0.5, 1, or 2 mg/kg/day of DHEA. After euthanization, beta-EP levels were measured in hippocampus, hypothalamus, anterior pituitary, neurointermediate pituitary, and plasma. Allopregnanolone and DHEAS levels were measured in hippocampus, hypothalamus, anterior pituitary, adrenal glands, and serum. Serum E(2) concentration was also measured. MAIN OUTCOME MEASURE(S): Dehydroepiandrosterone sulfate ester (DHEAS), E(2), beta-EP, and allopregnanolone levels. RESULT(S): Dehydroepiandrosterone administration increased DHEAS content in all organs and serum, except for anterior pituitary, where no significant changes occurred. DHEA administration in ovariectomized animals did not significantly increase E(2) circulating levels. DHEA administration induced an increase in allopregnanolone and beta-EP content in hippocampus, hypothalamus, and anterior pituitary and in serum or plasma. CONCLUSION(S): Dehydroepiandrosterone administration in ovariectomized animals increased allopregnanolone and beta-EP central and peripheral levels, which suggests that this compound may play a role as a neuroendocrine mediator, possibly substantiating the beneficial effects of postmenopausal DHEA therapy on the central nervous system. PMID- 15831290 TI - Studies of gene expression in human cumulus cells indicate pentraxin 3 as a possible marker for oocyte quality. AB - OBJECTIVE: To search for differentially expressed genes in cumulus cells from two groups of oocytes with different developmental outcome in vitro. DESIGN: Analyses of gene expression in human cumulus cells from oocytes that failed to fertilize in vitro (group A) and those that developed into normal-appearing embryos on day 3 (group B). SETTING: University-based facilities for clinical service and research. PATIENT(S): Women undergoing IVF treatment for infertility. INTERVENTION(S): Cumulus cells were collected from oocytes that were aspirated from ovarian follicles for IVF. The oocytes were cultured individually for IVF and embryo development. Total RNA was extracted from the cumulus cells for gene expression analyses. MAIN OUTCOME MEASURE(S): General gene expression profiles and relative abundance of pentraxin 3 (Ptx3) mRNA. RESULT(S): DNA microarray analysis identified 160 genes, including Ptx3, that were differentially expressed between cumulus cells in group A and group B. Quantitative analysis confirmed that the relative abundance of Ptx3 mRNA in cumulus cells was highly associated with oocyte development. CONCLUSION(S): This study demonstrated that changes in the expression levels of 160 genes, including particularly Ptx3, in human cumulus cells may be indicative of the quality of their enclosed oocyte. PMID- 15831291 TI - Time course of follicular development after bovine ovarian tissue transplantation in male non-obese diabetic severe combined immunodeficient mice. AB - OBJECTIVE: The aim of this study was to determine, with a bovine model, the appropriate interval for xenografted adult and newborn ovarian tissue to develop gonadotropin-responsive follicles. DESIGN: Controlled experiment. SETTING: Academic research laboratory. ANIMAL(S): Male non-obese diabetic (NOD) severe combined immunodeficient (SCID) mice (n = 20) were hosts of bovine ovarian tissue. Two dairy calves and one adult beef cow were donors of ovarian tissue. INTERVENTION(S): Newborn and adult bovine ovarian cortical pieces were transplanted to the SC space of intact male NOD SCID mice. Grafts were recovered after euthanasia at intervals after transplantation. MAIN OUTCOME MEASURE(S): Microscopic examination of histologic sections to determine proportions of growing follicles. RESULT(S): There was an increase in the proportion of primary and secondary follicles on day 55 after surgery for the cow and on day 124 after surgery for calf tissue compared with nongrafted and xenografted ovarian tissues recovered at previous intervals. These observed increases were accompanied by decreases in proportions of primordial follicles. Results suggest a sudden increase in the proportion of primary and secondary follicles due to progressive development of primordial follicles. CONCLUSION(S): In the NOD SCID mouse, bovine follicles survived xenotransplantation and underwent development. A longer interval was required for ovarian follicular development in calf tissues compared with that in adult cow ovarian tissues after xenotransplantation. PMID- 15831292 TI - Immunocytochemical detection and reverse transcription polymerase chain reaction expression of oncostatin M (OSM) and its receptor (OSM-Rbeta) in human fetal and adult ovaries. AB - OBJECTIVE: To investigate the immunocytochemical expression and presence of mRNA transcripts of oncostatin M (OSM) and its exclusive receptor (OSM-Rbeta) in ovaries from human adults and fetuses. DESIGN: Immunocytochemical and reverse transcription polymerase chain reaction (RT-PCR) study. SETTING: Major tertiary care and referral academic centers. PATIENT(S): Ten women and girls undergoing laparoscopic ovarian biopsy and 30 women undergoing second-trimester and third trimester pregnancy terminations. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Microscopic morphometric analysis, immunocytochemistry for OSM and OSM-Rbeta, and RT-PCR analyses. RESULT(S): There was strong to moderate immunocytochemical staining for OSM in both oocytes and granulosa cells of follicles from primordial stages onwards in ovaries from both fetuses and adults/adolescents. OSM-Rbeta was detected mainly in the oocytes. Transcripts of OSM and OSM-Rbeta RNA were detected by RT-PCR analyses. CONCLUSION(S): The expression of OSM and its receptor in ovarian tissue from fetuses and women suggests a possible role of OSM in growth initiation of human primordial follicles. PMID- 15831293 TI - Noninvasive imaging of spindle dynamics during mammalian oocyte activation. AB - OBJECTIVE: To develop a method to evaluate spindle dynamics in living oocytes and in karyoplasts during the initial stages of activation and after pharmacological disruption of cytoskeleton. DESIGN: Morphological study using a novel microscope. SETTING: Translational research laboratory at marine biological laboratory. ANIMAL(S): Six-week-old CD-1 or B6C3F1 mice superovulated with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG). INTERVENTION(S): Spindles of living oocytes and karyoplasts were imaged at 5-10 minute intervals using the Pol-Scope during the initial stages of oocyte activation and after pharmacological disruption of cytoskeleton. MAIN OUTCOME MEASURE(S): Assessment of spindle dynamics using Pol-Scope imaging. RESULT(S): During oocyte activation, spindle mid-region birefringence increased, followed by spindle rotation and second polar body extrusion in both intact oocytes and karyoplasts. Activation of protein kinase C (PKC) with phorbol 12-myristate 13-acetate failed to induce spindle activation in 60% of living oocytes and caused spindle disruption in some oocytes. Inhibition of PKC by a myristoylated PKC pseudosubstrate inhibited metaphase II release in most oocytes evaluated (86.7%). Cytochalasin D inhibited only spindle rotation and separation. Nocodazole disrupted spindles in less than 5 minutes after administration. CONCLUSION(S): Pol-Scope imaging allows investigation at near real time of spindle dynamics during activation of living oocytes. Spindles also showed evidence of activation even in karyoplasts. The procedure may be useful for detecting functional spindle aberrations in living oocytes. Further studies are needed to determine whether spindle dynamics predict clinical outcome. PMID- 15831294 TI - Differences in gene expression in the proliferative human endometrium. AB - OBJECTIVE: To use microdissection and DNA microarray technology to demonstrate differences in gene expression between epithelial and stromal areas in the proliferative human endometrium. DESIGN: Pilot study. SETTING: University hospital. PATIENT(S): Patients with normal menstrual cycles and at least one previous intrauterine pregnancy. INTERVENTION(S): Uterine endometrial biopsy. MAIN OUTCOME MEASURE(S): Gene expression. RESULT(S): From a total of 1,200 genes, 14 were strongly expressed in epithelial areas and 12 were strongly expressed in stromal areas. Among the genes strongly expressed in the stroma, expressions of decorin and discoidin domain receptor were confirmed by real-time polymerase chain reaction. Decorin was localized in the stromal areas by immunohistochemical staining. To confirm the effects of estrogen on gene expression, stromal cells were cultured. When E(2) was added to the culture media, expression of decorin mRNA was increased. CONCLUSION(S): The data demonstrated in this study help to understand the physiology of human endometrium. Decorin was strongly expressed in the stromal areas and was regulated by estrogen, and therefore it may be involved in restoration of the endometrium. PMID- 15831295 TI - Hypoxia upregulates cyclooxygenase-2 and prostaglandin E(2) levels in human peritoneal fibroblasts. AB - OBJECTIVE: To determine the levels of COX-1, COX-2, and prostaglandin (PG) E(2) in human fibroblasts isolated from normal peritoneal and adhesion tissues. DESIGN: Prospective experimental study. SETTING: University medical center. PATIENT(S): Fibroblast cultures from both peritoneum and adhesion tissues of five patients. INTERVENTION(S): Treatment of the primary cultured fibroblasts with NS398. MAIN OUTCOME MEASURE(S): We used Western blot to determine the effects of hypoxia on COX-1 and COX-2 levels from lysates of normal peritoneal and adhesion fibroblasts before and after hypoxia. We also used the ELISA techniques to determine PGE(2) levels in media collected from these fibroblasts before and after hypoxia and with and without NS398, a COX-2-specific inhibitor. RESULT(S): There was no difference in COX-1 levels between normal and adhesion fibroblasts with and without hypoxia. Basal COX-2 and PGE(2) levels were significantly higher in adhesion than normal fibroblasts. Hypoxia gradually increased COX-2 and PGE(2) levels in normal peritoneal fibroblasts over time, reaching a peak at 24 hours but had no effect on adhesion fibroblasts. Inhibition of COX-2 by NS398 significantly reduced PGE(2) levels in both normal and adhesion fibroblasts. CONCLUSION(S): The presence of higher levels of COX-2 in adhesion fibroblasts and the induction of COX-2 in normal peritoneal fibroblasts in response to hypoxia indicate a possible inflammatory response. Regulation of COX-2 may alter peritoneal healing and may provide the opportunity to reduce postoperative adhesion development. PMID- 15831296 TI - Oxytocin receptor expression in smooth muscle cells of peritoneal endometriotic lesions and ovarian endometriotic cysts. AB - OBJECTIVE: To investigate the expression of oxytocin receptor (OTR) in peritoneal and ovarian endometriotic lesions. DESIGN: Retrospective nonrandomized study. SETTING: University hospital endometriosis research center. PATIENT(S): Premenopausal women with histologically confirmed endometriosis were selected. Peritoneal endometriotic lesions (n = 120); ovarian endometriotic cysts (n = 40); peritoneal biopsies, distant from the endometriotic lesion (n = 55); and unaffected peritoneal biopsies from patients without endometriosis (n = 11) were obtained. Hysterectomy specimens from patients without endometriosis and/or adenomyosis were used for controls (n = 10). INTERVENTION(S): Histopathological examination of peritoneal and ovarian specimens for OTR expression and identification of smooth muscle cells by immunohistochemistry staining with antibodies against OTR and smooth muscle actin. In addition, Western blot analysis, double-immunofluorescence, and in vitro studies with primary cell cultures have been performed. MAIN OUTCOME MEASURE(S): Comparison of the immunoreactive score of the OTR and smooth muscle actin expression with the smooth muscle content in peritoneum with and without endometriosis. RESULT(S): In the epithelial cells of endometriotic lesions, we could demonstrate a high OTR expression. The stromal cells were OTR negative with the exception of some single cells. By using a monoclonal anti-smooth muscle actin antibody, these cells could be identified as intrastromal OTR-positive smooth muscle cells. The peritoneum of women with endometriosis shows a significantly higher smooth muscle content than the peritoneum of women without endometriosis. There were no significant differences between the smooth muscle content of active or inactive lesions and the stage of disease. CONCLUSION(S): Oxytocin receptor is expressed in smooth muscle cells and epithelial cells of peritoneal endometriotic lesions and ovarian endometriotic cysts. The inhibition of OTR by specific inhibitors might be a useful approach for the treatment of endometriosis-associated pain. PMID- 15831297 TI - Human leukocyte antigen expression by peritoneal macrophages from women with pelvic endometriosis is depressed but coordinated with costimulatory molecule expression. AB - OBJECTIVE: To investigate the macrophage response in endometriosis, we determined expression of human leukocyte antigen (HLA)-ABC, HLA-DR, and their costimulatory molecules by peritoneal fluid (PF) macrophages. DESIGN: Case-control study of immunologic markers. SETTING: University hospital. PATIENT(S): We compared 38 Japanese women with endometriosis with 59 control subjects who were given other laparoscopic diagnoses. INTERVENTION(S): Venipuncture and laparoscopic peritoneal fluid collection. MAIN OUTCOME MEASURE(S): Expression of HLA-ABC, HLA-DR, CD54, CD40, CD58, CD80, and CD86 by peripheral blood (PB) monocytes and PF macrophages was quantitated as mean fluorescence intensities by flow cytometry. Expression of each marker on PF macrophages was divided by that on PB monocytes as an index of macrophage activation (macrophage activation ratio). RESULT(S): In women with endometriosis, PF macrophages showed significant positive correlations between expression of HLA-ABC and other costimulatory molecules and also between HLA-DR and their costimulatory molecules. However, expression of HLA-ABC and DR by PF macrophages, and also their activation ratios, were significantly lower than in controls. CONCLUSION(S): Coordination with costimulatory molecules but relatively low expression of HLA-ABC and HLA-DR indicates a positive but limited immune response (antigen presentation) to events in the peritoneal cavity in women with endometriosis. This may induce immune tolerance to implanted or metaplastic endometrial tissue. PMID- 15831298 TI - Expression of steroid receptors, their cofactors, and aromatase in human luteinized granulosa cells after controlled ovarian hyperstimulation. AB - OBJECTIVE: To examine expression of androgen receptor (AR), AR cofactors, estrogen (E) receptor alpha, E receptor beta, progesterone receptor, steroid receptor coactivator-1, and aromatase in human luteinized granulosa cells collected during oocyte retrieval. DESIGN: Prospective real-time reverse transcriptase-polymerase chain reaction study. SETTING: Academic medical center. PATIENT(S): A total of 198 samples were brought into the study. INTERVENTION(S): Patients underwent the long protocol for assisted reproductive technology. Luteinized granulosa cells were collected transvaginally with ultrasound guidance. Quantitative reverse transcriptase-polymerase chain reaction was performed to quantify the mRNA expression of the investigated genes. MAIN OUTCOME MEASURE(S): The expression levels were determined as ratios between the studied genes and the reference gene beta-actin. RESULT(S): There is little AR expression in human luteinized granulosa cells immediately preceding ovulation under controlled ovarian hyperstimulation. All aspirated follicles, despite their antral size, displayed a similar mRNA expression of the investigated genes in the luteinized granulosa cells. CONCLUSION(S): This study supports the possibility of a transition of androgen action from being an enhancer of follicular differentiation (through the AR) to being a substrate of E synthesis (through aromatase) at the time of oocyte retrieval. The present study also demonstrates no effect of follicular size upon the status of steroid receptor mRNA expression in the luteinized granulosa cells when follicles were at least >1.5 mL. PMID- 15831299 TI - Coculture of monkey ovarian tissue increases survival after vitrification and slow-rate freezing. AB - OBJECTIVE: To assess whether coculture of monkey ovarian tissue after low temperature storage enhances follicular viability. To assess a novel method of vitrifying ovarian tissue. DESIGN: Prospective in vitro study. SETTING: University-affiliated national research center. ANIMAL(S): Ovaries from 15 cynomolgus or rhesus macaques (1-11 years). INTERVENTION(S): Vitrification using a containerless liquid nitrogen emersion system that involves dropping thin cortical pieces suspended in cyroprotectant directly into liquid nitrogen with outcome compared with slow-rate-controlled freezing. Before analysis, some of the thawed tissue was cocultured on mitotically inactivated mouse fetal fibroblast monolayers supplemented with FSH, insulin, transferrin, and selenium. MAIN OUTCOME MEASURE(S): Percentage of oocytes viable using live-dead fluorescent staining with carboxyfluorescein diacetate succinimidyl ester and propidium iodide. RESULT(S): Postthaw survival rates were 70.4% +/- 4.8% of 1,705 follicles after vitrification and 67.3% +/- 1.9% of 1,895 follicles after slow-rate freeze in six trials with each method. Coculture of the thawed tissue increased the viability, respectively, to 89% +/- 2.1% of 2,833 follicles previously vitrified and to 90.3% +/- 1.9% of 2,109 follicles after a slow-rate freeze (P<.01). Primordial follicles (30- to 50-microm diameter) were the vast majority of surviving follicles after thaw and coculture. Follicular viability in control fresh tissue (eight trials) was 76.0% +/- 4.1%, suggesting negligible loss in follicular viability after cryopreservation. CONCLUSION(S): Coculture of thawed ovarian tissue on mouse fetal fibroblasts and FSH increases the percentage of viable follicles. A novel method of vitrifying ovarian tissue is as effective as slow-rate freezing. These approaches may improve graft survival and function when used to treat chemotherapy-induced sterility. PMID- 15831300 TI - The antiprogesterone Org 31710 inhibits human blastocyst-endometrial interactions in vitro. AB - OBJECTIVE: To investigate the effect of the anti-P Org 31710 on human blastocyst attachment to cultured endometrial epithelial cells. DESIGN: Experimental in vitro study. SETTING: University hospital. PATIENT(S): Eleven fertile endometrial donors. INTERVENTION(S): Timed endometrial biopsy for cell cultures. MAIN OUTCOME MEASURE(S): Blastocyst attachment rate on endometrial cell cultures; secretion of glycodelin and leukemia inhibitory factor into the culture medium measured by RIA and ELISA techniques; and expression of progesterone receptors, interleukin-1 receptor type-1, and integrin subunit beta(3) on endometrial epithelial cells examined by immunohistochemistry. Endometrial pinopodes visualized by scanning electron microscopy. RESULT(S): Eleven of 16 human blastocysts attached to control cultures, whereas none of 10 blastocysts attached when Org 31710 was added to the culture medium (P=.0007). Immunohistochemical studies demonstrated no significant differences between groups. Biochemical analyses displayed a trend toward higher glycodelin secretions and, by scanning electron microscopy, a tendency toward less pinopode formation in the Org 31710 group, but the results did not reach statistical significance. The presence of swollen microvilli, precursors of endometrial pinopodes, was significantly reduced on cultures with Org 31710 (P=.03). CONCLUSION(S): The study presents a model for human blastocyst endometrial interactions responding to an anti-P drug. The exact mechanism for the anti-attachment properties of Org 31710 on the cultured endometrial cells and the blastocysts needs further evaluations. PMID- 15831301 TI - Development of a hamster superovulation program and adverse effects of gonadotropins on microfilament formation during oocyte development. AB - OBJECTIVE: To establish a superovulation procedure for the golden hamster (Mesocricetus auratus) by elucidating gonadotropin effects on oocyte development. DESIGN: Randomized, prospective study. SETTING: University laboratory of embryology and gamete biotechnology. ANIMAL(S): Twelve- to 15-week-old female and sexually mature male hamsters. INTERVENTION(S): Different doses of pregnant mare serum gonadotropin (PMSG) were injected into female hamsters in metestrus, diestrus, or proestrus. The same dose of hCG was injected 56 hours later. MAIN OUTCOME MEASURE(S): Embryo development and oocyte morphology after treatment. RESULT(S): First, 10 IU or 15 IU each of PMSG and hCG was injected into 10 hamsters weighing <110 or 110-130 g, respectively. All hamsters were mated, but none delivered live young after injection. Second, the doses of 15 IU, 7.5 IU, 5 IU, or 0 IU of each gonadotropin were injected into each hamster (regardless of body weight, 5 per each group). Increasing numbers of embryos were retrieved as the dosage was increased (11.2 to 46.6 embryos per hamster), whereas the percentage of two-cell embryos at retrieval was significantly decreased (100% to 3%, P<.05). In subsequent culture, none developed to blastocysts after 15-IU injection, whereas 47%, 55%, and 70% of two-cell embryos developed after 7.5-IU, 5-IU, and 0-IU treatments, respectively. As a result, females injected with 5 IU yielded more blastocysts than did females without injection (67 vs. 39). The number of inner cell mass cells per blastocyst was greatly increased in the control groups compared with the 5-IU and 7.5-IU treatment groups (22 vs. 14.3 14.7 cells per blastocyst). Third, the ultrastructure of oocytes was examined after injecting 5 IU each of PMSG and hCG (regardless of body weight). Superovulation did not affect oocyte maturation, but different patterns in microfilament formation were detected after the treatment. CONCLUSION(S): Female hamsters can be superovulated effectively by injecting equal amounts of PMSG and hCG, 56 hours apart. However, embryo development was adversely affected in a dose dependent manner at all doses of gonadotropins, and microfilament distribution was affected by such treatment. PMID- 15831302 TI - Crosslinked hyaluronan hydrogels containing mitomycin C reduce postoperative abdominal adhesions. AB - OBJECTIVE: To evaluate the efficacy of crosslinked hyaluronan (HA) hydrogels that contained covalently-bound mitomycin C (MMC) in reducing postoperative adhesions in a rat uterine horn model. DESIGN: Two independent parameters were investigated: [1] the quantity of MMC in preformed crosslinked hydrogel films and [2] the efficacy of intraperitoneal injection of in situ crosslinkable solutions. SETTING: University animal research facility. ANIMAL(S): Female Wistar rats. INTERVENTION(S): Injuries (3 x 10 mm) were made to contacting serosal surfaces of the medial uterine wall musculature in female rats. Two treatment protocols were used. In the first, sterile crosslinked HA films that contained different MMC loadings (0, 0.5%, and 2%) were applied to two injured uterine horns; control animals received no films. In the second protocol, MMC-loaded crosslinked HA gels that contained different MMC loadings (0.31%, 0.625%, and 1.25%) were spread on the site of uterine horn injury (1 mL); then, an additional 4 mL of the same formulation was injected into the peritoneal cavity after abdominal closure. Control animals were injected with 5 mL of buffer only. MAIN OUTCOME MEASURE(S): Extent of postoperative adhesions between uterine horns and with surrounding tissues and organs. RESULT(S): Mitomycin C-loaded crosslinked HA films and in situ crosslinked gels were more effective in reducing postoperative adhesion formation than were buffer controls or crosslinked HA films without MMC. CONCLUSION(S): Mitomycin C-loaded crosslinked HA films and gels reduced formation of postoperative intraperitoneal adhesions. PMID- 15831303 TI - Tubal sterilization with a waterborne polyethylene glycol in situ cross-linking material: a minimally invasive approach. AB - OBJECTIVE: To assess the effectiveness and compatibility of an in situ-gelling polymeric device when implanted in the fallopian tube to prevent pregnancy. DESIGN: Controlled laboratory study. SETTING: Animals in a controlled laboratory environment. ANIMAL(S): Ten white New Zealand rabbits (nine female and one male). INTERVENTION(S): The material was implanted through a small incision into one of each rabbit's twin fallopian tubes. The rabbits then were allowed to heal. After a healing period of 6 to 10 weeks, the rabbits were bred. After breeding, the animals were killed, and samples were collected from their fallopian tubes for histology. MAIN OUTCOME MEASURE(S): Pregnancy of the specimens. RESULT(S): Four specimens showed unilateral pregnancy (pregnancy in only one uterus). Because rabbits have the capacity to be pregnant in both uteri simultaneously, this suggests strong potential of the material for use as a contraceptive. CONCLUSION(S): Although concerns remain concerning distension and potential injury resulting from use of the system, enough data have been gathered to warrant further study of this material for use in humans. PMID- 15831304 TI - Identification of differential gene expression in germinal vesicle vs. metaphase II mouse oocytes by using annealing control primers. AB - By using a new innovative technology, annealing control primer-polymerase chain reaction (ACP-PCR), we identified genes that are differentially expressed in immature GV vs. in mature MII mouse oocytes. The results of the present study will be valuable in understanding the components of oocyte maturation and provide a basis for studies of human oocyte maturation. PMID- 15831305 TI - Differential expression of L-selectin ligand in the endometrium during the menstrual cycle. AB - OBJECTIVE: To evaluate the expression of L-selectin ligand in the endometrium during the menstrual cycle. DESIGN: Retrospective study. SETTING: University teaching hospital. PATIENT(S): Endometrial samples from regularly cycling women. INTERVENTION(S): Tissue microarray and immunohistochemical staining were performed using L-selectin ligand monoclonal antibody (MECA-79). MAIN OUTCOME MEASURE(S): Expression of L-selectin ligand in the various phases of menstrual cycle was measured by a semiquantitative analysis (HSCORE) for the intensity of immunohistochemical reactivity. RESULT(S): In the luminal epithelium, there were significant differences in L-selectin ligand expression during the proliferative, interval, early secretory, and midsecretory phases. The expression of L-selectin ligand was greatest from the periovulatory interval through midsecretory phase. In the glandular epithelium, the expression of L-selectin ligand was greatest in midsecretory phase with significant differences between proliferative phase and the midsecretory phase and between the interval phase and the midsecretory phase. CONCLUSION(S): Increased expression of L-selectin ligand in the human endometrium during the early and midsecretory phases of the menstrual cycle may be related to the process of implantation. PMID- 15831306 TI - Effects of long-acting recombinant human follicle-stimulating hormone analogs containing N-linked glycosylation on murine folliculogenesis. AB - OBJECTIVE: To evaluate the efficacy of two novel long-acting rhFSH analogs, rhFSH N2 and rhFSH-N4, in stimulating murine folliculogenesis. DESIGN: Experimental study. SETTING: Academic research environment. ANIMAL(S): Immature female mice. INTERVENTION(S): Recombinant hFSH-N2 and -N4 were administered via single IP injection to 3-week-old female mice (n = 10) who were killed 48 hours later for dissection and histologic examination of reproductive organs and serum inhibin A. Results were compared with other groups of mice who received either single or q 12 hour injections for 48 hours of commercial rhFSH, or a single injection of pregnant mare serum gonadotropin (PMSG). A subgroup of the mice receiving rhFSH N4 was supplemented with daily injections of small doses of hCG to simulate LH add-back. MAIN OUTCOME MEASURE(S): Serum inhibin A levels, ovarian and uterine weights, and ovarian antral follicle counts. RESULTS(S): Recombinant human FSH-N2 and -N4 administration induced a statistically significant increase in ovarian weights, uterine weights, and inhibin A levels compared with single and twice daily injection of rhFSH. PMSG induced the greatest increases in all three measured parameters. There was no statistical difference between rhFSH-N2 and rhFSH-N4 for any parameter analyzed. A single injection of rhFSH-N2 or -N4 induced a greater number of antral follicles than did either single or q 12 hour injections of rhFSH. The addition of small doses of hCG to rhFSH-N4 increased inhibin A levels and antral follicle number to reach statistical equivalence to PMSG treatment. CONCLUSION(S): Addition of a synthetic polypeptide containing two or four N-linked glycosylation sites to rhFSH increases in vivo bioactivity of the hormone compared to commercial rhFSH. After a single injection, both rhFSH-N2 and rhFSH-N4 effectively induced a greater follicular response in the mouse than did rhFSH. PMID- 15831307 TI - Comparison of two methods of in vivo gene transfer by electroporation. AB - OBJECTIVE: To evaluate two contrasting methods of in vivo gene transfer into testicular cells using electroporation, with regard to efficiency of transfer and damage to the testes. DESIGN: Controlled animal study. SETTING: Research laboratory at a university medical school. ANIMAL(S): 8-10-week-old male mice. INTERVENTION(S): The reporter construct pCAGGS-LacZ consisting of a cytomegalovirus enhancer/chicken beta-actin promoter attached to the LacZ gene was introduced into the testes in vivo using electroporation. For eight weeks, the efficiency and extent of LacZ gene expression, and the extent to which the testis was damaged by the technique, were investigated. MAIN OUTCOME MEASURE(S): Beta-galactosidase activity resulting from expression of the LacZ transgene was verified by X-gal staining, and LacZ mRNA expression was determined by RT-PCR analysis. Potential disorders associated with seminiferous tubular sperm formation were evaluated using the Johnsen score. RESULT(S): Long-lasting beta galactosidase activity was detected in spermatogenic cells up to eight weeks postelectroporation. Apparent damage to spermatogenesis was evident but was transient in nature and recovered with time; this plasticity was particularly evident following rete testes injection. CONCLUSION(S): Injection into the rete testis appears to be more suitable for in vivo gene transfer by electroporation than direct intratesticular injection. PMID- 15831308 TI - Effects of chronic hyperandrogenism and/or administered central nervous system insulin on ovarian manifestation and gonadotropin and steroid secretion. AB - OBJECTIVE: Polycystic ovarian syndrome is characterized by hyperandrogenism and insulin resistance. We studied the effects of central hyperinsulinemia and peripheral hyperandrogenism on gonadotropin secretion, steroid secretion, and ovarian histology in female rats. DESIGN: Experimental in vivo animal study. SETTING: University research center. ANIMAL(S): 250-300 g female Wistar rats. INTERVENTION(S): Insertion of testosterone pellets and/or administration of intracerebroventricular (ICV) insulin. MAIN OUTCOME MEASURE(S): Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels before and after GnRH administration; testosterone, insulin, and glucose levels; and ovarian histology. RESULT(S): Compared with control, rats with testosterone implant had a lower LH and higher FSH while rats with testosterone plus insulin had a higher FSH. Intracerebroventricular (ICV) insulin alone increased LH in response to GnRH. Ovarian histology indicated that testosterone-implanted rats had larger ovaries and an increased number of cystic follicles >50 microm as well as substantial theca enlargement. The administration of testosterone did not alter serum insulin, and ICV insulin did not increase testosterone levels. CONCLUSION(S): We suggest that ICV insulin acts either directly or indirectly to increase the LH responsiveness to GnRH. ICV insulin arrested the maturation of follicles leading to an increase in the number of small follicles. Peripheral androgens stimulated theca enlargement with cystic follicles. The combination of ICV insulin and peripheral androgens attenuated ovarian histologic changes and gonadotropin secretion. Thus, central hyperinsulinemia and peripheral hyperandrogenism may play a role in gonadotropin secretion as well as ovarian morphology. PMID- 15831310 TI - The elbow: diagnosis and treatment of common injuries. AB - This article deals with common injuries to the elbow. Elbow anatomy is reviewed. Diagnosis and treatment of pronator syndrome,lateral epicondylitis (tennis elbow), radial tunnel syndrome, posterior interosseous nerve syndrome, medial epicondylitis (golfer's elbow), ulnar collateral ligament injury, cubital tunnel syndrome,posterolateral rotatory instability, distal biceps injuries, tricepstendon injuries, and posterior elbow impingement are discussed. PMID- 15831311 TI - The wrist: clinical anatomy and physical examination--an update. AB - A pain-free, functional wrist joint is critical to many sporting and vocational activities. Assessing the painful wrist requires knowledge of common injury patterns and frequently seen conditions. The primary care physician with an understanding of basic wrist anatomy and biomechanics and a systematic approach to assessing the wrist will be more comfortable and effective with diagnosing and managing the athletically injured wrist. PMID- 15831312 TI - The wrist: common injuries and management. AB - Primary care physicians not only have an important role in the diagnosis and initial treatment of wrist injuries, but also play a key role in the education of families about prevention. Children and adolescents are often competitive in sports throughout the year. Periods of rest can be important in prevention of overuse injuries in the very active, developing athlete. Protective gear such as wrist guards, used during activities such as inline skating and snowboarding, has been shown to prevent acute injuries that often require surgery or lead to prolonged disability [84,85].A primary care physician will often be the first health care provider to assess most wrist complaints. The intent of this article is to familiarize the primary care physician with the most common wrist injuries in active people, and to demonstrate that many injuries can have poor outcomes if unrecognized. It is important to have good clinical knowledge of the functional anatomy of the wrist in order to maximize the information gathered on examination and to narrow one's differential diagnosis. The athlete's sport and desires regarding return to play, and the impact of the timing of injury management on his or her further participation in sport are important to consider. A highly active person may be referred to a musculoskeletal specialist for advanced testing or surgical repair earlier in the evaluation of certain injuries than a less active one. Armed with good clinical knowledge of anatomy and an understanding of common wrist injuries,primary care physicians can successfully manage many wrist complaints. PMID- 15831313 TI - The hand in sports: an update on the clinical anatomy and physical examination. AB - The hand plays an important role in the athlete's ability to compete, and there is a tendency to misdiagnose and undertreat these injuries. Thus,a detailed anatomic understanding and a timely and accurate diagnosis through a thorough physical examination are critical to guide the treatment and return of normal function of the hand. PMID- 15831314 TI - Hand injuries in sports medicine. AB - Many athletic injuries of the hand commonly present to, and can be managed by, primary care physicians. Knowing when to refer is also obviously important. This article is not meant to be an exhaustive list of all possible hand injuries. Rather, it is meant to serve as a useful guide to the primary care physician. For the busy primary care physician, a thorough assessment of the hand or wrist complaint includes a focused history and physical and the judicious use of supportive diagnostic testing. Know your anatomy and sport. This will set the foundation clinically for the best possible outcome. Ultimately,the goals of assessment and management are to restore, maximize, and preserve function. PMID- 15831315 TI - Clinical examination of the foot and ankle. AB - The foot and ankle are critical components in our ability to ambulate. Injuries to either can significantly interfere with a patient's ability to carry out normal activities. In severe cases, they can be devastating to a patient's independence. Careful examination of the foot and ankle using established mechanical tests, along with understanding of the anatomy of the complex,is needed to confirm the history and to assist in the diagnosis and treatment of foot and ankle injuries. The following points are key to clinical examination of the foot and ankle: . The examination of the foot and ankle needs to be done with the patient in both weight-bearing and non-weight bearing positions. . The examination of the foot and ankle should include an evaluation of the patient's gait. . Reproduction of a patient's symptoms is the key to making a correct diagnosis. . Although anatomic variants may predispose some individuals to injury,in general, if asymptomatic, no treatment should be done. PMID- 15831316 TI - Ankle and foot injuries in pediatric and adult athletes. AB - Foot and ankle injuries are among the most common in athletes.Differential diagnosis, imaging decisions, and treatment plan should be influenced by the age of the athlete. A thorough history and physical examination, with an understanding of the anatomy of the foot and ankle anatomy and the mechanism of injury, will give the best opportunity to make the correct diagnosis. For most athletes, the prognosis for returning to play after a foot or ankle injury is very good. The recipe for keeping our athletes healthy includes early intervention, undergoing proper rehabilitation, applying braces or orthotics when indicated, and preventing injuries when appropriate. PMID- 15831317 TI - Groin injuries and groin pain in athletes: part 1. AB - Sports injuries to the hip and groin region have been noted in 5% to 9% of high school athletes. These injuries occur most commonly in athletes participating in sports involving side-to-side cutting, quick accelerations and decelerations, and sudden directional changes.Symptoms may range from intermittent episodes of mild discomfort to severe and chronic career-ending pain. Groin injuries may result from a variety of causes. Although this article deals mainly with athletic etiologies, the physician must keep in mind that many other medical conditions may also affect the groin. Because of these overlapping medical conditions and because the anatomy of the region is so complex, a team approach is optimal. In Part 1 of this two-part series, ligament/muscle strains, nerve entrapment syndromes, and posterior abdominal wall abnormalities are reviewed as common causes of groin pain. PMID- 15831318 TI - Groin injuries and groin pain in athletes: part 2. AB - Sports injuries to the hip and groin region have been noted in 5% to 9% of high school athletes. These injuries occur most commonly in athletes participating in sports involving side-to-side cutting, quick accelerations and decelerations, and sudden directional changes.Symptoms may range from intermittent episodes of mild discomfort to severe and chronic career-ending pain. Groin injuries may result from a variety of causes. Although this article deals mainly with athletic etiologies, the physician must keep in mind that many other medical conditions may also affect the groin. Because of these overlapping medical conditions and because the anatomy of the region is so complex, a team approach is optimal. In the second of this two-part series, disorders of the os pubis, stress fractures and various hip pathologies are reviewed as causes of groin pain. PMID- 15831319 TI - Lower back pain in the athlete: common conditions and treatment. AB - Athletes younger than 12 years of age commonly have pathology related to the lower back pain. Spondylolysis is the most common condition in these athletes. Other conditions, including lumbar Scheuermann's disease,scoliosis, disc herniation, fractures, and muscular stains, can occur. Most of the mature general population experiences low back at some time in life. Athletes may be at increased risk, but outcomes are good. The majority of low back pain in mature athletes is mechanical in nature. Herniated discs,spinal stenosis, sacoilitis, and sacral stress fractures can also cause low back pain in these athletes. Low back conditions mentioned above may be treated with rest, specific exercise programs, and medication. Surgery is indicated for severe spinal stenosis, pain with evidence of neurological compromise,and some painful deformities. Newer treatments for back pain are emerging,but few controlled clinical trials are available. PMID- 15831320 TI - Cervical pain in the athlete: common conditions and treatment. AB - In summary, it is important for physicians dealing with neck pain in an athletic population to understand the differences between serious and mild cervical injuries. This is best facilitated by a thorough understanding of the signs and symptoms of serious cervical injury, familiarity with the basic anatomy of the neck and its structures, and a working knowledge of common causes of neck pain and mechanisms of injury. All unconscious athletes should be assumed to have a serious cervical injury until proven otherwise, and preventive measures should be taken to ensure the safety of the athlete. This includes airway management with a jaw thrust only, neck stabilization, and preventing helmet removal. In the conscious athlete who has neck pain, serious cervical injury can often be ruled out with an accurate history and physical examination. In all cases of neck pain, it is imperative that the athlete be protected. This may involve removing the athlete from competition, or transporting him or her to the local emergency room. Often, this decision falls on the shoulders of the doctor in the stands. Thus, a basic understanding of the evaluation and management of neck pain in athletes is an asset for all physicians who frequent athletic events or see athletes in the office. The physician responsible for patients who have Down syndrome or rheumatoid arthritis needs to consider the increased incidence of cervical instability in these patients when evaluating for athletic participation or neck pain. PMID- 15831321 TI - Basic biomechanics of the lower extremity. AB - This article describes common principles of biomechanics for running, jumping, and kicking. These activities form the basis for much of sports activity. Understanding human movement allows the physician to prescribe appropriate prevention, treatment, and rehabilitation to patients. PMID- 15831322 TI - Sports-related osteochondral injuries: clinical presentation, diagnosis, and treatment. AB - These lesions present with a variety of symptoms and historical findings.A careful joint-specific examination and appropriate imaging studies can help direct the physician in the treatment of the patient who has a chondral,osteochondral, or osteochondritis dissecans lesion. Categorizing the patient based on the size of the lesion, physiologic age, and desired activity level can aid in determining which patients might succeed with nonoperative versus operative management. PMID- 15831323 TI - Sports pharmacology and ergogenic aids. AB - Primary care physicians working with athletes need to ask about drug or supplement use. A basic knowledge of ergogenic substances may help to establish rapport with athletes using these agents. Physicians should be aware that doses used by athletes are far in excess of the ones reported by the literature, and that new substances are constantly being introduced. It is hoped that increased awareness and knowledge of the more common ergogenic substances will lead to better education and health care for the athletic population. PMID- 15831324 TI - A review of calibration techniques for freehand 3-D ultrasound systems. AB - Three-dimensional (3-D) ultrasound (US) is an emerging new technology with numerous clinical applications. Ultrasound probe calibration is an obligatory step to build 3-D volumes from 2-D images acquired in a freehand US system. The role of calibration is to find the mathematical transformation that converts the 2-D coordinates of pixels in the US image into 3-D coordinates in the frame of reference of a position sensor attached to the US probe. This article is a comprehensive review of what has been published in the field of US probe calibration for 3-D US. The article covers the topics of tracking technologies, US image acquisition, phantom design, speed of sound issues, feature extraction, least-squares minimization, temporal calibration, calibration evaluation techniques and phantom comparisons. The calibration phantoms and methods have also been classified in tables to give a better overview of the existing methods. PMID- 15831325 TI - 3-D endobronchial ultrasonography--a post mortem study. AB - Endobronchial ultrasonography (EBUS) using balloon-embedded probes may cause airway obstruction and requires a rapid scanning procedure. Three-dimensional (3 D) postprocessing of sequential 2-D images may allow detailed studies on a rapidly acquired image volume. We applied a 3-D reconstruction program on EBUS recordings acquired with a radial-scanning probe during a controlled pullback procedure in water-filled airways in five corpses. The aim was to acquire EBUS images under optimized conditions and to test a 3-D software program for postprocessing. Under post mortem conditions, 3-D images were obtained, allowing any-plane imaging and measurements, application of different rendering and visualization algorithms and merging of overlapping volumes. PMID- 15831326 TI - Estimating myocardial attenuation from M-mode ultrasonic backscatter. AB - The objective of this investigation was to determine whether measurements of myocardial attenuation can be obtained from analyses of M-mode images. We exploited the inherent anisotropy of myocardial properties as a means of systematically varying the attenuation to evaluate this M-mode image-based method for myocardial tissue characterization. A commercially available ultrasonic imaging system was used to acquire M-mode images of 24 excised cylindrical specimens from six formalin-fixed sheep hearts that were analyzed using video signal analysis. Data were compensated for the presence of bright intramural myocardial echoes, a potentially significant contributor to uncertainty in measurements of attenuation from backscattered ultrasound. The estimated attenuation coefficient in dB/cm at an effective center frequency of 2.75 MHz as a function of angle of insonification for measurements obtained from analyses of M-mode images is presented. Given a linear frequency-dependence of attenuation in myocardial tissue over frequencies ranging from 1.5 MHz to 8 MHz, as has been previously reported, M-mode image-based analyses were used to estimate the slope of attenuation. Results showed slopes of attenuation (over a -10 dB transmit bandwidth of 1.875 MHz to 3.75 MHz) ranging from 1.00 +/- 0.07 to 1.81 +/- 0.08 dB/(cm.MHz) for perpendicular and parallel insonification, respectively. These values were in good agreement with contemporaneously measured values (0.99 +/- 0.02 to 1.77 +/- 0.04 dB/(cm.MHz)) obtained over a frequency bandwidth of 4 MHz to 7 MHz using a through-transmission radio-frequency-based approach. These data suggest that robust measurements of myocardial attenuation can be obtained from analyses of M-mode images and that this method may be diagnostically feasible in the clinical setting. PMID- 15831327 TI - Ultrasonic sound velocity measurement in samples of soft materials through under resonance excitation. AB - Ultrasound (US) velocity determination is a valuable characterization technique, providing important information on elastic properties of materials. Sound velocity can be obtained accurately in the pulsed method if the thickness of the specimen is precisely known. This is clearly not easily achievable for soft materials, such as biologic soft tissues or tissue-mimicking phantoms. From this consideration, previous works have established that sound velocity can be determined in through-transmission configuration without thickness measurement through the time-of-flight determination of specimen-reflected echoes in plane parallel-surfaced specimens. It is shown here that the amplitude and shape of these specimen echoes can be significantly improved by working in the tone-burst mode at an excitation frequency below the transducer resonance. This is particularly valuable for materials presenting a low acoustic contrast with the surrounding medium, usually water, such as tissue-mimicking materials and water based phantoms, making the specimen echo time-of-flights and, consequently, the sound velocity determination, more reliable. PMID- 15831328 TI - RF and amplitude-based probe pressure correction for 3D ultrasound. AB - Anatomical deformation caused by variable probe contact pressure is a significant problem in freehand 3D ultrasound, particularly for high resolution musculoskeletal and breast scans. We have previously published an amplitude-based algorithm for correcting such errors. In this paper, we compare this approach with a novel, elastography-inspired algorithm which works with the higher resolution radio-frequency (RF) signal. The results show that, although the RF based algorithm is more precise in certain circumstances, both algorithms are able to compensate for probe pressure in 3D ultrasound data. Consequently, freehand 3D ultrasound users who do not have access to the RF signal are still in a position to perform effective probe pressure correction using the readily available video output, as long as this signal is not affected by significant amounts of frame averaging (persistence). PMID- 15831329 TI - A mechanical instrument for 3D ultrasound probe calibration. AB - We present a novel technique for 3D ultrasound probe calibration. The principle of operation is that the beam is aligned with a set of coplanar wires strung across a rigid frame. The probe and frame are mounted on a precision-manufactured mechanical instrument which allows adjustment and measurement of their relative pose. Semi-automatic image processing facilitates alignment of the beam and wires to within a tolerance of around 0.2 mm, despite the considerable beam thickness. The calibration process requires just a single view and relatively little user expertise. In a series of experiments with different ultrasound probes, we demonstrate the technique's high accuracy and precision. The latter is partly due to the elimination of the position sensor, a significant source of measurement noise, from the end-user calibration process. PMID- 15831330 TI - Real-time numerical simulation of Doppler ultrasound in the presence of nonaxial flow. AB - Numerical simulations of Doppler ultrasound (DUS) relying on computational fluid dynamics (CFD) models of nonaxial flow have traditionally employed detailed (but computationally intensive) models of the DUS physics, or have sacrificed much of the physics in the interest of computational or conceptual simplicity. In this paper, we present a compromise between these extremes, with the objective of simulating the essential characteristics of DUS spectrograms in a real-time manner. Specifically, a precomputed pulsatile CFD velocity field is interrogated at some number, N, of discrete points distributed spatially within a sample volume of prescribed geometry and power distribution and temporally within a prescribed sampling window. Intrinsic spectral broadening is accounted for by convolving each of the point velocities with a semiempirical broadening function. Real-time performance is facilitated through the use of an efficient algorithm for interpolating the unstructured CFD data. A spherical sample volume with Gaussian power distribution, N = 1000 sampling points, and quadratic broadening function are shown to be adequate for simulating, at frame rates of 86 Hz on a 1.5 GHz desktop workstation, realistic-looking spectrograms at representative locations within a stenosed carotid bifurcation model. Via qualitative comparisons with matched in vitro data, these simulated spectrograms are shown to mimic the distinctive spectral envelopes, broadening and power characteristics associated with common carotid, stenotic jet and poststenotic recirculating flows. We conclude that the complex interaction between Doppler ultrasound and complicated clinically relevant blood flow dynamics can be simulated in real time via this relatively straightforward semiempirical approach. PMID- 15831331 TI - Improvements in elastographic contrast-to-noise ratio using spatial-angular compounding. AB - Spatial-angular compounding is a new technique developed for improving the signal to-noise ratio (SNR) in elastography. Under this method, elastograms of a region of-interest (ROI) are obtained from a spatially weighted average of local strain estimated along different insonification angles. In this article, we investigate the improvements in the strain contrast and contrast-to-noise ratio (CNR) of the spatially compounded elastograms. Spatial angular compounding is also applied and evaluated in conjunction with global temporal stretching. Quantitative experimental results obtained using a single-inclusion tissue-mimicking phantom demonstrate that the strain contrast reduces slightly but the CNR improves by around 8 to 13 dB. We also present experimental spatial angular compounding results obtained from an in vitro thermal lesion in canine liver tissue embedded in a gelatin phantom that demonstrate the improved visual characteristics (due to the improved CNR) of the compound elastogram. The experimental results provide guidelines for the practical range of maximum insonification angles and estimates of the optimum angular increment. PMID- 15831332 TI - Property and contrast-enhancement effects of lipid ultrasound contrast agent: a preliminary experimental study. AB - This work investigated the influence of some factors on the property in vitro of a self-made lipid ultrasound (US) contrast agent (LCA) and evaluated the relationship of acoustic pressure and enhancement effect in normal rabbit kidney parenchyma. In the in vitro studies, filling gas, solvent and concentration of LCA solution were investigated. Morphologic characteristics, concentration and mean diameter of microbubbles were considered as indices. In the in vivo studies, contrast-tuned imaging (CnTi) technique was used to investigate the enhancement effects in kidney parenchyma under nine acoustic pressure levels. Among the samples saturated with different filling gases, perfluoropropane (C(3)F(8)) resulted in the highest concentration of microbubbles and air, the lowest. Microbubbles filled with C(3)F(8) or sulfur hexafluoride (SF(6)) were quite stable and remained at a high level of concentration (above 2 x 10(9) microbubbles per mL) much longer than did air-filled microbubbles. Among the four solutions tested, 5% glucose solution and 0.9% saline solution showed higher initial concentrations and greater longevity than dextran 40 glucose solution (6%) or distilled water. The concentration of LCA solution had a positive correlation with the microbubble concentration. All microbubble samples under different test conditions remained shape-complete and no aggregation or fusion was observed. The mean diameter of microbubble samples was about 3.4 microm. Contrast intensity and longevity of CnTi enhancement in vivo showed an acoustic pressure-dependent decrease. At 1 kPa, contrast intensity increased 224-fold (4.47/0.02) and the longevity of CnTi enhancement in the kidney parenchyma remained longer. PMID- 15831333 TI - Laser Doppler anemometry measurements of the shear stresses on ultrasonic contrast agent microbubbles attached to agar. AB - Ultrasonic contrast agents are currently being developed to target and bind to specific areas of interest such as atheromous plaque. A microbubble has been developed in-house which can be targeted to attach to specific cell-lines. To assess the feasibility of using the microbubble in vivo, the shear stresses which the bound microbubbles can withstand need to be known. A flow chamber was developed for use with intravascular ultrasound (IVUS) and laser Doppler anemometry (LDA). Biotin was incorporated into the microbubble shells and streptavidin was used to attach them to agar. IVUS at 40 MHz was then used to image the attached microbubbles under steady flow at a range of flow rates from 75 to 480 mL min(-1) through a flow area of 9 mm(2). LDA was employed to find high resolution velocity profiles of the flow in the chamber at a selection of these flow rates and the shear stresses on the bubbles were calculated. The bubbles were found to remain attached to the agar for shear stresses of up to 3.4 Pa. This compares with mean physiological arterial shear stresses of less than 1.5 Pa for pulsatile flow. PMID- 15831334 TI - Vascular effects induced by combined 1-MHz ultrasound and microbubble contrast agent treatments in vivo. AB - Previous in vivo studies have demonstrated that microvessel hemorrhages and alterations of endothelial permeability can be produced in tissues containing microbubble-based ultrasound contrast agents when those tissues are exposed to MHz-frequency pulsed ultrasound of sufficient pressure amplitudes. The general hypothesis guiding this research was that acoustic (viz., inertial) cavitation, rather than thermal insult, is the dominant mechanism by which such effects arise. We report the results of testing five specific hypotheses in an in vivo rabbit auricular blood vessel model: (1) acoustic cavitation nucleated by microbubble contrast agent can damage the endothelia of veins at relatively low spatial-peak temporal-average intensities, (2) such damage will be proportional to the peak negative pressure amplitude of the insonifying pulses, (3) damage will be confined largely to the intimal surface, with sparing of perivascular tissues, (4) greater damage will occur to the endothelial cells on the side of the vessel distal to the source transducer than on the proximal side and (5) ultrasound/contrast agent-induced endothelial damage can be inherently thrombogenic, or can aid sclerotherapeutic thrombogenesis through the application of otherwise subtherapeutic doses of thrombogenic drugs. Auricular vessels were exposed to 1-MHz focused ultrasound of variable peak pressure amplitude using low duty factor, fixed pulse parameters, with or without infusion of a shelled microbubble contrast agent. Extravasation of Evans blue dye and erythrocytes was assessed at the macroscopic level. Endothelial damage was assessed via scanning electron microscopy (SEM) image analysis. The hypotheses were supported by the data. We discuss potential therapeutic applications of vessel occlusion, e.g., occlusion of at-risk gastric varices. PMID- 15831335 TI - Thermal assessment of 40-MHz pulsed Doppler ultrasound in human eye. AB - Tissue exposure to diagnostic pulsed Doppler ultrasound (US) can cause significant temperature rises. Temperature rise induced by US biomicroscopy (UBM) system (VS40, VisualSonics, Toronto, ON, Canada) was measured in ex vivo human and rabbit eyes with a 26-gauge K-type needle thermocouple. The operating frequency was 40 MHz with a free field I(SPTA) of 2.6 mW/cm(2) (B-mode) and 11.9 W/cm(2) (Doppler). Peak negative pressures were 5.22 MPa (B-mode) and 7.32 MPa (Doppler), resulting in MIs of 0.83 (B-mode) and 1.05 (Doppler mode). In Doppler mode, mean temperature rises of 2.27 degrees C and 1.93 degrees C were measured for the human lens and ciliary body after a 3-min insonation, vs. 2.66 degrees C for the rabbit lens. Our results indicate that US-induced temperature rise decreases with decreasing number of cycles, decreasing pulse-repetition frequency (PRF) or increased transmit attenuation, and is consistent with simple models of heating. To limit risk of temperature rises of 1 degrees C in human ciliary body, use of the maximum settings of 16 cycles (0.400 micros pulse duration), 20-kHz PRF should include 3-dB transmit attenuation, and exposure time should be limited. For insonation of the lens, exposure settings no higher than nine cycles (0.225-micros pulse duration) and 10-kHz PRF should be employed and exposure time limited to minimize risk of temperature increases of 1 degree C. PMID- 15831336 TI - A new scanning approach for limb extremities using a water bag in freehand 3-D ultrasound. AB - 3-D ultrasound (US) can significantly improve the visualization of musculoskeletal tissues, such as residual limbs, feet and hands. Traditionally, mechanical scanning is normally required to obtain the entire volume of these limb extremities. In this paper, a new scanning approach using a water bag was described to collect the complete volume of various tissues surrounding bones. The water bag was used to contain the limb extremity and the scanning was conducted on its external surface from different directions. The recorded 2-D US images containing complete anatomic information surrounding the bones from different directions were used to form full 3-D volumes of the limb extremities. A plastic auxiliary apparatus was designed to hold the water bag and support the subject's limb part with an armrest. A corresponding algorithm was proposed to remove invalid image information within each sweep by a separating plane defined semiautomatically. Two phantoms were used to test the repeatability and accuracy of the imaging. The distance between two plastic bands attached to a plastic tube filled with US gel measured by a micrometer and from the four reconstructed volumes were 39.03 +/- 0.36 mm and 39.2 +/- 0.5 mm, respectively. The diameter, height and volume of a silicone cylinder phantom measured for the 10 reconstructed volumes were 40.2 +/- 1.4 mm, 12.9 +/- 1.0 mm and 16400 +/- 1600 mm(3), respectively. They agreed with the corresponding results obtained by the micrometer, which were 41.29 +/- 0.13 mm, 12.98 +/- 0.17 mm and 17370 +/- 140 mm(3), respectively. The reconstructed volumes of the two phantoms, a chicken leg in vitro, and human fingers in vivo were also reported. The preliminary results obtained in this study demonstrated that this new scanning approach should have potential for the 3-D US imaging of musculoskeletal extremities using freehand scanning. PMID- 15831337 TI - Secondhand smoke exposure in the workplace. AB - BACKGROUND: Currently, there is little understanding of the relationship between the strength of workplace smoking policies and the likelihood and duration, not just the likelihood, of exposure to secondhand smoke at work. METHODS: This study assessed self-reported exposure to secondhand smoke at work in hours per week among a cross-sectional sample of 3650 Massachusetts adults who were employed primarily at a single worksite outside the home that was not mainly outdoors. The sample data were from a larger longitudinal study designed to examine the effect of community-based tobacco control interventions on adult and youth smoking behavior. Participants were identified through a random-digit-dialing telephone survey. Multiple logistic regression and zero-inflated negative binomial regression models were used to estimate the independent effect of workplace smoking policies on the likelihood and duration of exposure to secondhand smoke. RESULTS: Compared to employees whose workplace banned smoking completely, those whose workplace provided designated smoking areas had 2.9 times the odds of being exposed to secondhand smoke and 1.74 times the duration of exposure, while those with no restrictions had 10.27 times the odds of being exposed and 6.34 times the duration of exposure. CONCLUSIONS: Workplace smoking policies substantially reduce the likelihood of self-reported secondhand smoke exposure among employees in the workplace and also greatly affect the duration of exposure. PMID- 15831338 TI - Organizational systems to support publicly funded tobacco treatment services. AB - BACKGROUND: Professional societies and government organizations have promoted guidelines and best practices that encourage clinicians to routinely integrate cessation counseling into patient encounters. While research in health maintenance organizations has demonstrated that the development and maintenance of office systems do enable clinicians' smoking-cessation services, little is known about the adoption of system strategies in diverse organizations serving disadvantaged populations. METHODS: Data were collected via face-to-face interviews from November 2001 to October 2002 using a standardized systems assessment checklist at service delivery sites of 83 funded community health service agencies, which included hospitals, community health centers, and other organizations (e.g., substance abuse, mental health, and multiservice). The content of the structured assessment reflected system elements with proven effectiveness that have been included in guidelines and best practices recommendations. Detailed information was collected on the implementation strategies. RESULTS: This study found considerable attention to systems that support cessation services in diverse healthcare organizations, but much remains to be done. There is a wide diversity of implementation strategies employed, with varied degrees of sophistication. CONCLUSIONS: A major challenge is to develop systems capable of providing population-based feedback to, and between, providers, which will enable further quality improvement efforts. PMID- 15831339 TI - Alcohol-impaired driving among U.S. adults, 1993-2002. AB - BACKGROUND: Motor vehicle-related injury is the leading cause of death in the United States for people aged 1 to 34 years. In 2002, 17,419 (41%) of 42,815 traffic deaths were alcohol related. OBJECTIVE: To estimate trends in alcohol impaired driving among U.S. adults from 1993 through 2002. DESIGN, SETTING, AND PARTICIPANTS: The Behavioral Risk Factor Surveillance System, a random-digit telephone survey of adults aged >/=18 years in all states (and the District of Columbia). MAIN OUTCOME MEASURES: The percentage of respondents who reported alcohol-impaired driving (AID) in the past month, total estimated annual number of AID episodes, and annual rate per 1000 adult population. RESULTS: The estimated annual number of episodes of AID in the United States declined from 123 million in 1993 to 116 million in 1997, but then increased to 159 million in both 1999 and 2002. In varying magnitudes, this increase was observed among most subgroups of the population. In each study year, over 80% of total AID episodes were reported by people who also reported binge drinking (more than five drinks on a single occasion). CONCLUSIONS: After a general decline in the United States in the mid-1990s, self-reported AID increased substantially by the turn of the century. AID is strongly associated with binge drinking. Effective interventions to prevent AID and binge drinking should be widely adopted. PMID- 15831341 TI - Playground safety and access in Boston neighborhoods. AB - BACKGROUND: Youth physical activity is partly influenced by access to playgrounds and recreational opportunities. Playgrounds in disadvantaged areas may be less safe. METHODS: Investigators assessed safety at 154 playgrounds in Boston between July 2000 and July 2001. Playgrounds were geocoded and safety scores assigned to census block groups (CBGs). For each of Boston's 591 CBGs, investigators calculated the total number youth and proportions of black residents, adults without a high school degree, and youth living in poverty. Investigators assigned each CBG a safety score, and calculated distance from the CBG centroid to the nearest playground and nearest "safe" playground (top safety quartile). Statistical analyses were completed using SAS PROC GENMOD by October 2002. RESULTS: In bivariate analysis, playground safety was inversely associated with total CBG youth population (p =0.001) and proportions of black residents (p <0.001), youth in poverty (p =0.003) and residents with no high school degree (p <0.002). The proportion of black residents in the CBG was inversely associated with safety (p =0.013), independent of CBG educational attainment and numbers of youth. The average distance was 417 meters to the nearest playground and 1133 meters to the nearest "safe" playground. Distance to the nearest playground was inversely associated with the proportion of residents with no high school degree (p <0.0001) after controlling for numbers of youth and proportion of black residents. CBGs with more youth had greater distances to the safest playgrounds (p =0.04). CONCLUSIONS: In Boston, playground safety and access to playgrounds varied according to indicators of small-area socioeconomic and racial/ethnic composition. PMID- 15831340 TI - Cancer screening practices among cancer survivors. AB - BACKGROUND: Cancer survivors are more vulnerable to future cancers than individuals without cancer. As such, it is important to understand whether survivors are engaging in cancer screenings. METHODS: The screening practices reported in response to the 2000 Health Interview Survey of 2151 individuals with cancer were examined and compared to those of 30,195 individuals without cancer. RESULTS: The proportion of cancer survivors obtaining screenings ranged from 21% to 77%. Compared to individuals without cancer, women with cancer were more likely to have had a mammogram (odds ratio [OR]=1.8, 95% CI=1.5-2.1), a clinical breast exam (OR=2.2, 95% CI=1.9-2.5), and/or a Papanicolaou test (OR=1.3, 95% CI=1.1-1.5) in the recommended timeframe. Similarly, men with cancer were more likely than men without cancer to have had a prostate-specific antigen test performed (OR=2.5, 95% CI=2.0-3.0). All cancer survivors were more likely than individuals without a cancer diagnosis to have had a total body skin exam (OR=4.0, 95% CI=3.5-4.6), a fecal occult blood test (OR=1.4, 95% CI=1.2-1.6), and/or a colorectal exam (OR=2.2, 95% CI=1.9-2.5). Similar results were obtained when individuals diagnosed with the cancer for which the screen was designed to detect were excluded. CONCLUSIONS: The results demonstrate that cancer survivors have higher screening rates than individuals without a cancer diagnosis. Despite this, the proportion of survivors obtaining screenings varies considerably by the type of screen. An understanding of the impact of cancer screening in cancer survivors, as well as the reasons for and against obtaining cancer screenings, is necessary. PMID- 15831342 TI - Associations of body mass index with meniscal tears. AB - BACKGROUND: Meniscal tears are common knee injuries, with limited reported data on associated factors, let alone risk factors. The objective of this study was to determine whether associations exist between increasing obesity and meniscal tears leading to surgery. METHODS: We performed frequency-matched case-control studies using surgical case data for years 1996 to 2000 from administrative databases of two large Utah hospitals; each case was matched with three controls from a large cancer screening trial. Meniscal tear cases (262 male and 282 female) were determined by surgical procedures. Inclusion criteria were age (50 to 79) and body mass index (BMI) (17.00 to 54.99 kg/m(2)). Gender-specific, age adjusted odds ratios with 95% confidence intervals (CIs) were calculated for BMI categories from <20.00 to >/=40.00. The referent BMI category was 20.00 to 22.49. RESULTS: Age-adjusted odds ratios for likelihood of meniscal surgery among those with a BMI of >/=40.00 were 15.0 (95% CI=3.8-59.0) for men, and 25.1 (95% CI=10.3 60.8) for women. All odds ratios for men and women with BMIs of >/=27.50 and >/=25.00, respectively, were statistically significantly elevated. CONCLUSIONS: Significant associations were demonstrated between increasing BMI and meniscal surgeries in both genders, including obese and overweight adults. PMID- 15831343 TI - Cardiovascular risk factors and confounders among nondrinking and moderate drinking U.S. adults. AB - BACKGROUND: Studies suggest that moderate drinkers have lower cardiovascular disease (CVD) mortality than nondrinkers and heavy drinkers, but there have been no randomized trials on this topic. Although most observational studies control for major cardiac risk factors, CVD is independently associated with other factors that could explain the CVD benefits ascribed to moderate drinking. METHODS: Data from the 2003 Behavioral Risk Factor Surveillance System, a population-based telephone survey of U.S. adults, was used to assess the prevalence of CVD risk factors and potential confounders among moderate drinkers and nondrinkers. Moderate drinkers were defined as men who drank an average of two drinks per day or fewer, or women who drank one drink or fewer per day. RESULTS: After adjusting for age and gender, nondrinkers were more likely to have characteristics associated with increased CVD mortality in terms of demographic factors, social factors, behavioral factors, access to health care, and health related conditions. Of the 30 CVD-associated factors or groups of factors that we assessed, 27 (90%) were significantly more prevalent among nondrinkers. Among factors with multiple categories (e.g., body weight), those in higher-risk groups were progressively more likely to be nondrinkers. Removing those with poor health status or a history of CVD did not affect the results. CONCLUSIONS: These findings suggest that some or all of the apparent protective effect of moderate alcohol consumption on CVD may be due to residual or unmeasured confounding. Given their limitations, nonrandomized studies about the health effects of moderate drinking should be interpreted with caution, particularly since excessive alcohol consumption is a leading health hazard in the United States. PMID- 15831344 TI - Ethnoracial differences among outdoor workers in key sun-safety behaviors. AB - BACKGROUND: Few sun-safety studies have included ethnoracially diverse groups. Comparison across such groups of sun-safety behaviors was the main objective of this analysis. METHODS: Postal workers (n =2543) self-reported frequency of sunscreen, wide-brim hat, and sunglasses use during the last 5 workdays on 5 point Likert-type scales ranging from "never" to "always." Responses were dichotomized by "always" versus all other responses. Generalized estimating equations were used to assess the association between ethnicity/race and frequency of each sun safety behavior. The baseline data reported in this paper were collected in the summer of 2001, and the randomized trial was completed in the summer of 2004. RESULTS: Ethnicity/race was significantly related to sunscreen and sunglasses use. For both sunscreen and sunglasses, the white group had significantly higher "always" rates than the other four groups (30% vs 14% to 23% and 63% vs 44% to 52%, respectively). CONCLUSIONS: The results of this study indicated that although non-Latino white employees tended to have higher rates of sun-safety behaviors, the rates of adequate protection of all groups were low. PMID- 15831345 TI - Potential effects of the next 100 billion hamburgers sold by McDonald's. AB - BACKGROUND: McDonald's has sold >100 billion beef-based hamburgers worldwide with a potentially considerable health impact. This paper explores whether there would be any advantages if the next 100 billion burgers were instead plant-based burgers. METHODS: Nutrient composition of the beef hamburger patty and the McVeggie burger patty were obtained from the McDonald's website; sales data were obtained from the McDonald's customer service. RESULTS: Consuming 100 billion McDonald's beef burgers versus the same company's McVeggie burgers would provide, approximately, on average, an additional 550 million pounds of saturated fat and 1.2 billion total pounds of fat, as well as 1 billion fewer pounds of fiber, 660 million fewer pounds of protein, and no difference in calories. CONCLUSIONS: These data suggest that the McDonald's new McVeggie burger represents a less harmful fast-food choice than the beef burger. PMID- 15831346 TI - Methodologies for realizing the potential of health impact assessment. AB - Health impact assessment (HIA), a systematic assessment of potential health impacts of proposed public polices, programs, and projects, offers a means to advance population health by bringing public health research to bear on questions of public policy. The United States has been slow to adopt HIA, but considerable strides have been made in many other countries, and under the auspices of the World Health Organization and World Bank. Varied applications in these diverse milieu have given rise to diverse approaches to HIA-quantitative/analytic, participatory, and procedural-each with distinct disciplinary foundations, goals, and methodologies. Suitability of these approaches for different applications and their challenges are highlighted, along with areas in which methodologic work is most needed and most likely to advance the field from theory and infrequent application to more routine practice in the United States. PMID- 15831347 TI - Applying educational gaming to public health workforce emergency preparedness. AB - From natural disasters to terrorism, the demands of public health emergency response require innovative public health workforce readiness training. This training should be competency-based yet flexible, and able to foster a culture of professional and personal readiness more traditionally seen in non-public health first-response agencies. Building on the successful applications of game-based models in other organizational development settings, the Johns Hopkins Center for Public Health Preparedness piloted the Road Map to Preparedness curriculum in 2003. Over 1500 employees at six health departments in Maryland have received training via this program through November 2004. Designed to assist public health departments in creating and implementing a readiness training plan for their workforce, the Road Map to Preparedness uses the core competencies of the Centers for Disease Control and Prevention for all public health workers as its basic framework. PMID- 15831348 TI - Occupational medicine: the case for reform. AB - The specialty of occupational medicine is in peril, in large part because of its reliance on financing by industry, which has powerful incentives to limit costs and to favor physicians who are useful to their employers. Occupational physicians generally practice within the framework of the workers' compensation system. Serious flaws in the incentive structure of workers' compensation constrain objectivity in their practice. Under present law they are unavoidably subject to perverse influences from employers and insurance companies. A fundamental reform of workers' compensation law and practice is urgently needed to separate occupational physicians from the control of employers and workers' compensation insurers, whose interests should not be allowed to override the physicians' integrity or to compromise the specialty. PMID- 15831349 TI - Residency training in preventive medicine: challenges and opportunities. PMID- 15831350 TI - Treating obesity-related hypertension with surgery. PMID- 15831351 TI - Circadian blood pressure variation in morbidly obese hypertensive patients undergoing gastric bypass surgery. AB - BACKGROUND: Impaired blood pressure (BP) variation, often found in obesity and hypertension, is associated with increased cardiovascular risk. The effect of obesity surgery on BP variation is unknown. We performed this study to investigate the effect of gastric bypass surgery on circadian BP variation in morbidly obese hypertensive subjects. METHODS: The study group consisted of eight patients (mean age 35.0 +/- 9.1 years), with impaired circadian BP rhythm. Controls were eight well-matched subjects (34.6 +/- 9.8 years) with normal BP rhythm. All patients underwent gastric bypass surgery. The 24-h ambulatory BP measurements were performed before (baseline) and 8 weeks after gastric bypass surgery. RESULTS: Mean body weight in the study group and controls decreased from baseline of 129.0 +/- 21.9 kg and 134.1 +/- 27.2 kg to 116.7 +/- 21.1 kg (by 9.5% +/- 2.1%) and 121.6 +/- 25.7 kg (by 9.3% +/- 1.7%) after the surgery, respectively (P < .0001 v baseline). Mean 24-h, daytime, and night-time BP values decreased significantly and similarly in both groups. In the study group mean 24 h systolic and diastolic BP decreased from 154.7 +/- 12.3 mm Hg and 105.6 +/- 8.1 mm Hg to 138.5 +/- 9.3 mm Hg and 90.8 +/- 6.7 mm Hg and in controls from 158.2 +/ 16.5 mm Hg and 106.2 +/- 10.0 mm Hg to 136.3 +/- 10.8 mm Hg and 92.9 +/- 6.3 mm Hg (P < .0001 v baseline), respectively. Circadian BP rhythm in the study group returned to the normal profile (mean systolic nocturnal fall increased from 4.0% +/- 2.4% to 16.4% +/- 4.0% and diastolic, from 5.1% +/- 3.0% to 17.3% +/- 5.5%; P < .0001 v baseline), whereas it remained normal in the controls. CONCLUSIONS: Surgery-induced body weight loss in morbidly obese hypertensive subjects with impaired circadian BP variation is not only associated with BP reduction, but also with the restoration of normal BP rhythm. PMID- 15831352 TI - Insulin resistance in men with treated hypertension at increased risk for cardiovascular disease: results of a 3-year study. AB - BACKGROUND: Insulin resistance is an independent risk factor for cardiovascular disease. The aim of this research was to establish the frequency of insulin resistance in a normoglycemic, hypertensive, male population to determine whether insulin resistance could be predicted by assessment of conventional risk factors and to assess the rate of deterioration. METHODS: Insulin resistance was calculated in hypertensive patients and healthy control subjects using a homeostasis model assessment for insulin resistance (HOMA-IR) and was correlated with routinely measured clinical variables. RESULTS: A total of 106 patients and 18 healthy control subjects were recruited. The healthy control subjects were similar to the patients in age and body mass index (BMI) but had a lower fasting glucose, insulin, and HOMA-IR. Of the patients, 36 were insulin sensitive (IS, HOMA-IR <1.5) and 21 were insulin resistant (IR, HOMA-IR >3.0). Mean blood pressure and type and number of antihypertensive medications did not differ between the groups. Compared with IS patients, IR patients were younger, had a higher BMI, higher triglycerides, and lower HDL, but there was substantial overlap of these risk factors between the groups. More IR than IS patients developed an impaired fasting glucose after 1 year (48% v 11%, P < .001). After 3 years of follow-up, 10% of the IR group and none from the IS group were confirmed as having type 2 diabetes mellitus. CONCLUSIONS: In this study, insulin resistance was common (20%) and could not be reliably diagnosed using conventional risk factors. One-half of the IR patients developed an impaired fasting glucose by 1 year, and, of these individuals, 20% were confirmed as having type 2 diabetes mellitus by 3 years. PMID- 15831353 TI - Relation of genetic predisposition and insulin resistance to left ventricular hypertrophy in hypertension. AB - BACKGROUND: The aim of the study was to determine whether genetic predisposition to hypertension and insulin resistance are related to left ventricular (LV) hypertrophy in essential hypertension. METHODS: The study included 72 nondiabetic patients with essential hypertension and 15 normotensive control (NC) subjects. The 72 patients were divided into two groups according to genetic predisposition to hypertension. The family history (FH)(+) group included 33 patients with at least one essential hypertensive parent or sibling. The FH(-) group included 39 patients with weak genetic predisposition to hypertension. Insulin resistance was estimated using the homeostasis model assessment (HOMA). Echocardiographically determined LV mass (LVM) and relative wall thickness (RWT) were measured as markers of LV hypertrophy. RESULTS: The HOMA values in the FH(+) group (2.00 +/- 0.89) were significantly higher than those in either the FH(-) group (1.21 +/- 0.44) or NC subject group (0.91 +/- 0.24). The HOMA values in the FH(-) group were significantly higher than those in NC subjects. The LVM and RWT were greatest in the FH(+) group, followed by those in the FH(-) group and NC subjects. There were no significant differences in LVM and RWT between the FH(-) group and NC subjects. By multivariate analysis, HOMA value (P = .0011), male sex (P = .0032), body mass index (P = .0061), systolic blood pressure (P = .0245), and genetic predisposition to hypertension (P = .0441) remained determinants of LVM in nondiabetic patients with essential hypertension. CONCLUSIONS: Genetic predisposition to hypertension and the HOMA value appear to have additive impact on LV hypertrophy. This relation is independent of well-known determinants of LVM such as male sex, overweight, and high blood pressure. PMID- 15831354 TI - Prescribing patterns for antihypertensive drugs after the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial: report of experience in a health maintenance organization. AB - BACKGROUND: The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) reported that primary cardiovascular outcomes were the same in hypertensive patients treated with thiazide-type diuretics as in those treated with calcium channel blockers or angiotensin-converting enzyme inhibitors. The aim of this study was to assess prescribing patterns of antihypertensive agents in a health maintenance organization (HMO) before and after the publications of ALLHAT results. METHODS: Our analysis used computer-stored information from the pharmacy system of the HMO for the period between January 1, 1996, and December 31, 2003. The study assessed prescribing patterns for antihypertensive drugs in six classes: thiazide-type diuretics, other-type antihypertensive diuretics, angiotensin-converting enzyme inhibitors/angiotensin receptor blockers, calcium channel blockers, beta-blockers, and alpha-blockers. The monthly total number and relative percentage of dispensed prescriptions for each class of antihypertensive agent, including new prescriptions and refills, were calculated. RESULTS: The use of alpha-blockers for all prescriptions began to decrease after the closure of the alpha-blocker (doxazosin) arm of ALLHAT in January 2000. Prescriptions for the thiazide-type diuretics immediately increased after the ALLHAT publication in December 2002. During the first 6 months of 2003, the percentages of thiazide type diuretics were statistically significantly higher compared with the predicted values. This pattern held for all as well as for new antihypertensive prescriptions. CONCLUSIONS: The ALLHAT publications had large effects on the antihypertensive prescribing patterns in our population. Prescription of thiazide type diuretics significantly increased after the ALLHAT publication. Our findings establish that the response of physicians to new clinical evidence can be very rapid. PMID- 15831355 TI - Arterial stiffness evaluated by measurement of the QKD interval is an independent predictor of cardiovascular events. AB - BACKGROUND: The QKD interval is the time between the QRS wave on the electrocardiogram (ECG) and the detection of the last Korotkoff sound during BP measurement. Measurement of the QKD interval during ambulatory blood pressure (BP) monitoring provides an automatic evaluation of arterial stiffness. The objective of this longitudinal study was to examine the relationship between arterial stiffness assessed by this method and the occurrence of cardiovascular complications in a cohort of initially untreated hypertensive patients. METHODS: The initially untreated hypertensive patients were included in the cohort of hypertensives recruited to our center between January 1992 and December 1999. All benefited from ambulatory measurement of the QKD interval and BP over 24 h. Most of these patients were given antihypertensive treatment and were followed up by their family physicians, who were unaware of the QKD results. Their outcome was evaluated in 2003 by inquiring of the patients themselves or of their family physicians. RESULTS: The initial population comprised 412 patients. At the end point, 33 had been lost to follow-up. We recorded 49 cardiovascular events in this population with an average follow up of 65 +/- 36 months. In a Cox model, the arterial stiffness assessed by the QKD(100-60) as a percentage of the height predicted normal value was significantly associated with the occurrence of cardiovascular complications, independent of age and mean 24-h BP. CONCLUSIONS: This study using ambulatory measurement of the QKD interval showed that an increase in arterial stiffness was a marker of cardiovascular risk. We show for the first time that its value persists after taking account of the mean systolic pressure over 24 h. PMID- 15831356 TI - Determinants of the second derivative of the finger photoplethysmogram and brachial-ankle pulse-wave velocity: the Ohasama study. AB - BACKGROUND: We examined different characteristics of the second derivative of the finger photoplethysmogram (SDPTG) and brachial-ankle pulse wave velocity (BAPWV) for assessing arterial function in a large general population, along with the alteration of SDPTG in hypertension. METHODS: Finger SDPTG and BAPWV were measured in 848 (544 normotensive and 304 untreated hypertensive) subjects 34 to 88 years of age. For assessing SDPTG, we calculated the B:A and D:A ratios and the aging index (AGI), based on the height of the wave components. Using univariate and multivariate analyses, determinants of SDPTG indices and BAPWV as well as differences between normotensive and hypertensive subjects were evaluated. RESULTS: We found that BAPWV was independently and positively correlated with age, blood pressure (BP), heart rate (HR), and hemoglobin A1c. The D:A ratio and AGI showed a positive and the B:A ratio a negative independent correlation with age and BP. In contrast, the D:A ratio and AGI showed a negative and the B:A ratio a positive correlation with HR. The SDPTG indices, but not BAPWV, were independently associated with gender. The multivariate-adjusted D:A ratio, AGI, and BAPWV were significantly higher and the adjusted B:A ratio was lower in hypertensive than in normotensive subjects, and hypertensive subjects showed two-fold greater adjusted risks of having high D:A and low B:A ratios. The SDPTG indices and BAPWV were mildly correlated with each other in normotensive but not in untreated hypertensive subjects. CONCLUSION: Although the SDPTG depends on various factors in a manner different from BAPWV, it may be useful for detecting vascular aging accelerated by hypertension. PMID- 15831357 TI - Differential effects of angiotensin II on atherogenesis at the aortic sinus and descending aorta of apolipoprotein-E-deficient mice. AB - Angiotensin (Ang) II infusion increases atherosclerosis and leads to the formation of abdominal aortic aneurysms in apolipoprotein E-deficient (ApoE-/-) mice. Conversely, blockade of the renin-angiotensin system (RAS) decreases atherosclerosis in this model. However, there are conflicting data in the literature concerning responses to both Ang II infusion and RAS blockade which may depend on age, sex, dose, duration of treatment, and the site at which lesion area was measured. In the present study we examined the effects of Ang II infusion on lesion formation in male ApoE-/- mice both at the aortic sinus and in the descending aorta, starting at different ages, and varying in duration. We also tested the effects of the Ang II receptor antagonist losartan at different doses in both males and females. Blood pressure and plasma renin concentration (PRC) were measured as indicators of the hemodynamic and neurohormonal effects of these treatments. Administration of Ang II increased lesion area much more in the descending aorta than at the aortic sinus. However, spontaneous lesion development at the aortic sinus was much greater than in more distal regions of the aorta. Aneurysms were observed in all treatment groups but were less severe in animals treated from 4 weeks age, possibly because of protective remodeling. Losartan treatment reduced lesion area at the aortic sinus, although differences were only significant in female mice. These findings demonstrate regional and temporal differences in the sensitivity of the aorta to the effects of RAS stimulation and blockade, and may help to explain some of the discrepancies between previous reports from other laboratories. PMID- 15831358 TI - Angiotensin II type 2 receptors contribute to vascular responses in spontaneously hypertensive rats treated with angiotensin II type 1 receptor antagonists. AB - BACKGROUND: Vasoconstrictive, proliferative and oxidative effects of angiotensin II (Ang II) are mediated by Ang II type 1 (AT1) receptors. The effects of Ang II via the Ang II type 2 (AT2) receptor subtype (AT2R) are less well defined. Growing evidence shows the existence of cross-talk between the Ang II receptor subtypes, which is revealed by AT1R blockade. Hence, under certain conditions, AT2R may act as an antagonistic system with respect to the AT1R. METHODS: The present study was designed to investigate the effects of long-term treatment with the AT1R antagonist losartan on the AT2R-mediated response to Ang II in thoracic aortas isolated from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. Untreated animals from both groups were used as controls. The mRNA expression of AT1R and AT2R was measured by reverse transcription-polymerase chain reaction. RESULTS: During contraction in response to norepinephrine, Ang II induced concentration-dependent relaxation only in aortas isolated from SHR chronically treated with losartan (8 weeks; 30 mg/kg/day in drinking water). These relaxations were inhibited by the selective AT2R blocker PD123319, N(G) nitro-L-arginine methyl ester (L-NAME), and B2receptor antagonist HOE-140. Accordingly, nitric oxide (NO) production was increased by Ang II only in the aortas of treated SHR. After AT1R blockade, AT2R mRNA was significantly increased. These findings demonstrate that, in hypertensive rats, chronic AT1R blockade is associated with an inverted vasomotor response to Ang II via AT2R mediated NO production. CONCLUSIONS: The losartan-unmasked AT2R-vasorelaxation could significantly contribute to the beneficial hemodynamic effects of AT1R blockade. In view of this, our study highlights the importance of the integrated Ang II receptor network, which may help to define further the mechanisms of the well-established vascular protective effects of AT1R blockers. PMID- 15831360 TI - Aging is associated with increased matrix metalloproteinase-2 activity in the human aorta. AB - BACKGROUND: Aging is a major risk factor for the development of arterial stiffness and vascular disease, and it is related to the upregulation of matrix metalloproteinase-2 (MMP-2) in the aorta of rats and nonhuman primates. This study aimed to determine whether MMP activity in the human vasculature changes with aging. We also assessed regional differences in MMP activity at two locations in the arterial tree, the aorta and the internal mammary artery (IMA). METHODS: Both MMP-2 and MMP-9 activity in the human aorta and IMA were determined by gelatin zymography and were localized within the tissue using in situ zymography. Tissue inhibitor of metalloproteinase-2 (TIMP-2) levels was determined by Western blot. RESULTS: Active MMP-2 (but not pro-MMP-2, pro-MMP-9, or active MMP-9) was positively correlated with age in the human aorta (r = 0.65; P < .001) but not in the IMA. Active MMP-2 and TIMP-2 (but not pro-MMP-2 or pro- or active MMP-9) levels are higher in the aorta than in the IMA (P < .001; P < .05). In the aorta, MMP activity is highest in the intima and is also detectable in the media and adventitia. To a lesser extent, MMP activity is present in all layers of the IMA. CONCLUSIONS: This study demonstrates that age-related MMP-2 upregulation occurs in the human aorta but not in the IMA. PMID- 15831359 TI - Effects of dietary salt loading on the responses of isolated rat mesenteric arteries to leptin. AB - BACKGROUND: Leptin induces relaxation of vascular smooth muscle through an endothelium-dependent release of nitric oxide (EDNO) and administration of a high salt diet reduces the relaxation of vessels to EDNO. We would, therefore, predict that salt loading would reduce the leptin-induced dilatation. However, in salt loaded animals the relaxation to acetylcholine is maintained through an endothelial-dependent hypopolarizing factor instead of EDNO. These experiments were, therefore, designed to examine whether in salt-loaded animals the response to leptin would be reduced or whether, as for acetylcholine, an alternative mechanism would be substituted. METHODS: Weanling rats were given diets containing either 0.4% (n = 10) or 8% (n = 9) sodium chloride for 4 weeks. They were then killed and a length of second order mesenteric artery was mounted in a perfusion myograph with diameter changes measured using a microscope-video tracking system. The vessels were preconstricted with norepinephrine and then the effects of graded concentrations of leptin were determined. RESULTS: In vessels from the low salt animals leptin caused a dose-dependent dilatation (maximum change 31.4% +/- 5.8% of the initial norepinephrine-induced constriction) but in the high salt animals the change was only 3.4% +/- 1.1%. The nitric oxide synthase blocker Nomega-nitro-L-arginine methyl ester (L-NAME) abolished the responses, although responses could still be obtained in vessels from both groups to the NO donor, sodium nitroprusside. CONCLUSIONS: These results indicate that salt loading to rats almost completely abolishes the vasodilatation to leptin. This implies endothelial disruption and, unlike the response to acetylcholine, no other vasodilator mechanism is implicated. This could provide a link between high salt intake and hypertension because the known increase in sympathetic activity caused by leptin would not be countered by a direct vasorelaxation. PMID- 15831361 TI - Physical activity, plasma antioxidant capacity, and endothelium-dependent vasodilation in young and older men. AB - BACKGROUND: Sedentary aging is associated with oxidative stress and endothelial dysfunction. The aim of this study was to evaluate the relationship between long term physical activity, plasma antioxidant status, and conduit artery endothelial function in young and older healthy men. METHODS: In young (n = 16) and older athletes (n = 16) and in matched healthy sedentary subjects, endothelium dependent flow-mediated dilation (FMD) and endothelium-independent response to glyceryl trinitrate (GTN), 400 microg, were measured in the brachial artery from high-resolution ultrasonography. Plasma malondialdehyde (MDA) and antioxidant capacity as total oxyradical scavenging capacity (TOSC) were also evaluated. RESULTS: We found that FMD was lower (< or =0.01) in sedentary older subjects (2.3% +/- 1.0%) as compared with older athletes (5.3% +/- 3.2%) and both sedentary (5.4% +/- 2.0%) and athletically trained (6.1% +/- 3.2%) young subjects. Sedentary older subjects showed higher (P < or = .05) MDA levels and lower (P < .0001) plasma antioxidant capacity as compared with the other subgroups, whereas in older athletes MDA levels and antioxidant capacity were similar to those observed in the young subgroups. In the whole group, FMD, but not GTN, was negatively related to age (r = -0.31, P < .05) and directly related (P < or = .01) to VO2max (r = 0.49) and TOSC against peroxyl (r = 0.69) and hydroxyl radicals (r = 0.53). In the multivariate analysis, TOSC against peroxyl radicals resulted as the most significant predictor of FMD (R2 = 0.60; P = .003). CONCLUSIONS: These results suggest that regular physical activity is associated with preserved antioxidant defenses and endothelial function in older individuals. PMID- 15831362 TI - Association of interleukin-6 gene G-174C polymorphism and plasma plasminogen activator inhibitor-1 level in Chinese patients with and without hypertension. AB - BACKGROUND: The interleukin-6 (IL-6) gene promoter G-174C polymorphism has been associated with insulin resistance, hypertension, and coronary artery disease; however, its relationship with plasma PAI-1 level has not yet been studied. METHODS: The G-174C genotypes and plasma PAI-1 antigen and activity were determined in 424 Chinese subjects, 207 with hypertension and 217 without, to study the possible effects of IL-6 genotypes on the regulation of PAI-1 and blood pressure. RESULTS: Hypertensive patients showed significantly greater percentage of IL-6 GG genotype (51.7% v 33.2%, P < .001) and G allele frequency (71.7% v 59%, P < .001) than normotensive subjects. The GG genotypic group had significantly higher plasma PAI-1 activity (16.1 +/- 9.8 v 12.3 +/- 7.5 IU/mL, P = .03) and antigen (32.4 +/- 23.2 v 23.2 +/- 13.5 ng/mL, P = .01) than the CC genotypic group, with intermediate values in the GC genotypic group (15.9 +/- 9.0 IU/mL and 29.1 +/- 17.5 ng/mL). Multiple linear regression analysis in all study subjects and in normotensive subjects documented an independent dominant effect of IL-6 G-174C gene polymorphism on plasma levels of PAI-1 activity (P = .02 and .01) and antigen (P = .02 and .03) after log transformation and adjustment for confounding factors. CONCLUSIONS: The present study showed a positive association of the IL-6 GG genotype with hypertension and with elevated plasma PAI-1 level in normotensive individuals in a Chinese population in Taiwan. Our findings suggest that the IL-6 gene promoter G-174C polymorphism may affect the regulation of PAI 1 and blood pressure through an inflammatory mechanism. PMID- 15831363 TI - Association of C825T polymorphism of G protein beta3 subunit with the autonomic nervous system in young healthy Japanese individuals. AB - BACKGROUND: The T allele of the C825T polymorphism of the G protein beta3 subunit gene (GNB3) is reported to be associated with increased intracellular signal transduction and the prevalence of essential hypertension. Because the two major receptors in the autonomic nervous system (ANS), the adrenergic and muscarinic acetylcholine receptors, are G protein-coupled receptors, it was expected that the GNB3 C825T polymorphism was associated with ANS function. In the present study, we have investigated the association of this polymorphism with ANS in young, healthy Japanese male individuals. METHODS: A total of 94 young, healthy subjects underwent the genotyping for the GNB3 C825T polymorphism and electrocardiogram R-R interval power spectral analysis in supine rest and standing positions. RESULTS: There were no significant differences among genotypes in any of the characteristics investigated (body mass index, blood pressure, plasma glucose, insulin, lipids, and family history of hypertension, diabetes, or obesity). However, in power spectral analysis of heart rate variability, the very-low-frequency component when standing was higher in TT carriers than in CC carriers, and TT and CT carriers had a significantly higher sympathetic nervous system (SNS) index and lower parasympathetic nervous system (PNS) index when standing than CC carriers. In addition, we found that TT carriers showed no chronological variations in either SNS or PNS index after postural change. CONCLUSIONS: These observations suggested that GNB3 C825T polymorphism is associated with ANS in youth. These findings raise the possibility that individuals who are T allele carriers are at increased risk for developing hypertension in relation to ANS function. PMID- 15831364 TI - Gender differences of renal CYP-derived eicosanoid synthesis in rats fed a high fat diet. AB - BACKGROUND: Renal cytochrome P450 (CYP)-derived eicosanoids, hydroxyeicosatetraenoic acids (HETEs), epoxyeicosatrienoic acids (EETs), and dihydroxyeicosatrienoic acids (DHETs), have been shown to affect renal function and blood pressure (BP). We recently reported that high fat (HF) diet treatment in male rats increases BP and decreases production of these eicosanoids in the kidneys. However, at what level the downregulation of renal eicosanoid synthesis occurs and whether the HF diet has any effects on the regulation of renal eicosanoid synthesis in female rats are not known. The purpose of this study was to determine renal CYP-derived eicosanoid synthesis and its association with BP regulation in HF male and female rats. METHODS: In the first set of experiments, male and female rats were fed the HF or control diet for 10 weeks. In the second set of experiments, male and female rats were fed the HF diet for 10 days. In the third set of experiments, HF-fed and control female rats were treated with 5alpha dihydrotestosterone for 4 weeks. After treatment, BP, urinary sodium, sodium balance, eicosanoid production, and CYP enzyme expression were determined. RESULTS: An elevation of BP and a decrease of renal cortical eicosanoid production were found in HF male rats, but no BP and eicosanoid production changes were observed in HF female rats. The HF treatment also caused a significant decrease of eicosanoid production and a decrease of CYP4A and 2C23 expression in the proximal tubules of HF male rats. Moreover, the HF diet treatment in male rats caused an increase in cumulative sodium balance and an elevation of BP, whereas no change in cumulative sodium balance and BP was observed in female rats. The treatment of 5alpha-dihydrotestosterone increased BP and 20-HETE production in the renal microvessels, but had no effect on urinary sodium excretion and renal microvessel EET production in both control and HF-fed female rats. CONCLUSIONS: These results demonstrate that there are gender specific differences in regulation of renal eicosanoid synthesis, sodium balance, and BP caused by HF treatment, and it appears that androgens play some role in upregulation of renal microvessel 20-HETE production in both HF and control female rats. PMID- 15831365 TI - Possible involvement of aminopeptidase A in hypertension and renal damage in Dahl salt-sensitive rats. AB - BACKGROUND: Although aminopeptidase A (APA), which is abundant in the kidneys, is responsible for metabolizing angiotensin II (Ang II), its association with salt sensitivity remains uncertain. We aimed to clarify the involvement of APA in salt induced hypertension and renal damage. METHODS: Male Dahl salt-sensitive (DS) and Dahl salt-resistant (DR) rats were fed low-salt (0.3%) or high-salt diet (8%) from 6 weeks of age for 12 weeks. Tail-cuff-measured blood pressure (BP), renal APA activity, renal Ang II levels, histologic renal damage, and APA immunoreactivity were periodically examined. RESULTS: Systolic BP progressively increased only in DS rats given the high-salt diet (DS-8% rats). The DR-8% rats had approximately 3-fold higher renal APA activity than the rats given the low salt diet (DR-0.3% rats) during the maintenance on the high-salt diet. However, although DS-8% rats also had 2.5-fold higher renal APA activity than DS-0.3% rats at 10 weeks, continuing the high-salt diet afterward suppressed the activity in DS-8% rats below the levels observed in DS-0.3% rats. High-salt diet reduced renal Ang II levels by 30% in DR rats, whereas it showed a small and nonsignificant decrease in DS rats. The number of injured glomeruli was markedly elevated in DS-8% rats after 10 weeks. The APA immunostaining in DS-8% rats was enhanced in glomeruli displaying mild damage, diminished in the severely injured glomeruli, and absent in lesions with hyalinization. CONCLUSIONS: High-salt diet in DS rats increased renal APA activity, although renal injury remained mild, but then reduced it along with the progression of glomerulosclerosis, suggesting that reduced APA activity may be involved in the deterioration of salt-induced hypertension and renal injury. PMID- 15831366 TI - Increased renal production of angiotensin II and thromboxane B2 in conscious diabetic rats. AB - BACKGROUND: The mechanisms involved in development of cardiovascular complications associated with diabetes mellitus are not well elucidated. Among the vasoactive factors that may play a role in development of these complications are angiotensin II and thromboxane B2 (TXB2). We hypothesized that diabetes increases renal production of TXB2 through stimulation of angiotensin type-1 receptor. METHODS: We used a microdialysis technique to monitor changes in renal interstitial fluid (RIF) TXB2 in conscious streptozotocin-induced diabetes rat model. The RIF levels of angiotensin II and TXB2 were monitored before and during 6 weeks after development of diabetes and during treatment with the angiotensin type-1 receptor blocker valsartan at 10 mg/kg. Measurement of the urinary albumin excretion (UAE) was used to monitor the development and progression of diabetic nephropathy. RESULTS: The UAE was 81.62 +/- 1.31 ng/min, 184.75 +/- 9.41 ng/min (P < .01), and 229.84 +/- 4.49 ng/min (P < .0001) at baseline, week 3, and week 6, respectively, after induction of diabetes. Basal levels of RIF angiotensin II were 4.28 +/- 0.02 pg/mL and significantly increased to 6.24 +/- 0.31 pg/mL (P < .001) and 7.66 +/- 0.05 pg/mL (P < .001) at 3 and 6 weeks after development of diabetes. Similarly, basal RIF TXB2 was 197 +/- 27 pg/mL and increased to 488 +/- 80 pg/mL (P < .01) and 703 +/- 130 pg/mL (P < .01) at 3 and 6 weeks after development of diabetes. Valsartan caused further increase in RIF angiotensin II levels. In contrast, valsartan decreased RIF TXB2 levels at baseline to 85 +/- 11 pg/mL (P < .01), at 3 weeks to 141 +/- 17 pg/mL (P < .01), and at 6 weeks to 255 +/- 45 pg/mL (P < .01) after development of diabetes. CONCLUSIONS: These results demonstrate that diabetes mellitus is accompanied by increased renal production of angiotensin II and TXB2. The increase in TXB2 is mediated through stimulation of angiotensin type-1 receptor. PMID- 15831367 TI - PPARgamma in endothelial cells influences high fat diet-induced hypertension. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands improve human hypertension. However, the mechanism and site of this effect remains unknown, confounded by PPARgamma expression in many cell types, including endothelial cells (ECs). METHODS: To evaluate the vascular role of PPARgamma we used a conditional null mouse model. Specific disruption of PPARgamma in ECs was created by crossing Tie2-Cre+ transgenic (T2T+) and PPARgamma-floxed (fl/fl) mice to generate PPARgamma (fl/fl)T2T+ (PPARgamma E-null) mice. Conscious 8- to 12 week-old congenic PPARgamma (fl/fl)Cre- (wild type) and PPARgamma E-null mice were examined for changes in systolic blood pressure (BP) and heart rate (HR), untreated, after 2 months of salt-loading (drinking water), and after treatment for 3 months with high fat (HF) diet alone or supplemented during the last 2 weeks with rosiglitazone (3 mg/kg/d). RESULTS: Untreated PPARgamma E-nulls were phenotypically indistinguishable from wild-type littermates. However, compared to similarly treated wild types, HF-treated PPARgamma E-nulls had significantly elevated systolic BP not seen after normal diet or salt-loading. Despite sex dependent baseline differences, salt-loaded and HF-treated PPARgamma E-nulls of either sex had significantly elevated HR versus wild types. Interestingly, rosiglitazone improved serum insulin levels, but not HF diet-induced hypertension, in PPARgamma E-null mice. CONCLUSIONS: These results suggest that PPARgamma in ECs not only is an important regulator of hypertension and HR under stressed conditions mimicking those arising in type 2 diabetics, but also mediates the antihypertensive effects of rosiglitazone. These data add evidence supporting a beneficial role for PPARgamma-specific ligands in the treatment of hypertension, and suggest therapeutic strategies targeting ECs may prove useful. PMID- 15831368 TI - Potential role of angiotensin-converting enzyme inhibitors and statins on early podocyte damage in a model of type 2 diabetes mellitus, obesity, and mild hypertension. AB - BACKGROUND: Experimental findings suggest that the obese Zucker rat (OZR) is a model of type 2 diabetes-related nephropathy with several metabolic abnormalities. However, the exact mechanisms by which these factors cause early glomerulosclerosis and proteinuria remain unclear. Furthermore, structural abnormalities and regulation of podocytes have recently emerged as prominent underlying factors in proteinuria. The aim of this study was to evaluate the potential role of angiotensin-converting enzyme inhibitors and statins on early podocyte damage in an experimental model of type 2 diabetes mellitus. METHODS: We used OZR to evaluate some of the pathogenic mechanisms and the effects of two drugs, an angiotensin-converting enzyme (ACE) inhibitor (quinapril) and a statin (atorvastatin), involved in the development of proteinuria and especially podocyte damage. We studied glomerular and tubulointerstitial injury by assessing inflammation mediators (murine monoclonal antibody against CD68 [ED1+], interleukin-8 [IL-8], interferon-gamma-inducible protein 10 [IP-10]) and podocyte damage markers using desmin staining and electron microscopy. RESULTS: Glomerular lesions were correlated with cholesterol (r = 0.676), proteinuria (r = 0.804), triglycerides (r = 0.593), insulin (r = 0.345), creatinine (r = 0.266), and glucose (r = 0.245). In addition, podocytes from OZR showed positive staining for desmin. Use of the ACE inhibitor quinapril normalized proteinuria, cholesterol levels, glomerular lesions, and podocyte morphology. In contrast, atorvastatin ameliorated but did not normalize renal damage, with a partial reduction in desmin staining and podocyte morphology. Treatment with both drugs resulted in only a slight reduction in IL-8 and IP-10 in the tubulointerstitium. CONCLUSIONS: In the OZR, cholesterol was an important determinant of renal injury. Most notably, glomerulosclerosis in the OZR Is characterized by early podocyte damage and tubulointerstitial injury. In addition, our findings showed that quinapril primarily normalized podocyte morphology, whereas atorvastatin ameliorated renal lesions through the diminution of lipids and by its lipid-independent pleiotropic effect. PMID- 15831369 TI - Self-reported hypertension treatment beliefs and practices of primary care physicians in a managed care organization. AB - BACKGROUND: Blood pressure (BP) is controlled to recommended goal in less than one-third of people with hypertension. There has been little recent research on physician beliefs and practices with regard to the treatment of hypertension. METHODS: In late 1999, we surveyed 104 primary care physicians in the 18 owned clinics of a large staff model, non-profit health maintenance organization. The survey included questions about demographics, BP treatment goals for patients with uncomplicated hypertension, and beliefs about hypertension. RESULTS: The reported systolic BP treatment goal was < or =140 mm Hg for 97% and the diastolic BP goal was < or =90 mm Hg for 100%. The systolic BP goal for patients with isolated systolic hypertension was < or =140 mm Hg for 82%, but 34% stated that they would treat to a different goal depending on the diastolic BP. The proportions of physicians who would intensify treatment for BP of 140/90 mm Hg, 150/95 mm Hg, 165/75 mm Hg, and 165/65 mm Hg were 64%, 97%, 89% and 77%, respectively. Although 93% believed that medication was necessary to control BP in most cases, a majority (55%) agreed with the statement that BP could be controlled in most patients with only one drug. Although 42% reported that they often had to change drugs because of side effects, only 16% believed that it was time-consuming to find a well-tolerated drug regimen. CONCLUSIONS: In this setting, primary care physicians' self-reported practices were in good agreement with national guidelines put forth in the late 1990s, and their beliefs were favorable to therapy. Our data point to a need for interventions to emphasize that combination drug therapy is frequently required to achieve BP control, and that more aggressive intervention is often warranted for isolated systolic hypertension. PMID- 15831371 TI - JNC-7 versus renin-based strategies for optimal antihypertensive drug treatment. PMID- 15831372 TI - Alpha-blocker monotherapy and anticipated occurrences of heart failure in hypertensive veterans. PMID- 15831374 TI - Loss of susceptibility as an alternative for nematode resistance. AB - Among plant pathogens, sedentary endoparasitic nematodes are one of the most damaging pests in global agriculture. These obligate parasites interact with their hosts in a quite unique and intriguing way. They induce the redifferentiation of root cells into specialized feeding cells essential for nematode growth and reproduction; thus, nematodes have evolved the ability to exploit plant genes and hijack host functions for their own requirements. Various approaches to engineer plants with resistance to parasitic nematodes have been pursued, most focusing on the introduction of resistance genes. An alternative strategy to achieve resistance is to exploit the susceptibility of plant disease. Better knowledge of the plant response during the compatible interaction should allow the identification of targets to engineer resistance to parasitic nematodes in crop species. PMID- 15831375 TI - Resistance to plant viruses: old issue, news answers? AB - Viruses represent significant threats to modern agriculture and their biological control remains a challenge in the twenty-first century. Recent progress has been made in our understanding of natural and engineered virus resistance, the two major strategies used for crop protection. The molecular mechanisms underlying the roles of both dominant and recessive resistance genes have been elucidated, promoting the development of possible new antiviral strategies. Engineering resistance in plants, in particular RNA-mediated protection, is also becoming increasingly important and is likely to play a key role in the future. PMID- 15831376 TI - Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. AB - Abiotic stresses, especially salinity and drought, are the primary causes of crop loss worldwide. Plant adaptation to environmental stresses is dependent upon the activation of cascades of molecular networks involved in stress perception, signal transduction, and the expression of specific stress-related genes and metabolites. Consequently, engineering genes that protect and maintain the function and structure of cellular components can enhance tolerance to stress. Our limited knowledge of stress-associated metabolism remains a major gap in our understanding; therefore, comprehensive profiling of stress-associated metabolites is most relevant to the successful molecular breeding of stress tolerant crop plants. Unraveling additional stress-associated gene resources, from both crop plants and highly salt- and drought-tolerant model plants, will enable future molecular dissection of salt-tolerance mechanisms in important crop plants. PMID- 15831377 TI - Phytoremediation: novel approaches to cleaning up polluted soils. AB - Environmental pollution with metals and xenobiotics is a global problem, and the development of phytoremediation technologies for the plant-based clean-up of contaminated soils is therefore of significant interest. Phytoremediation technologies are currently available for only a small subset of pollution problems, such as arsenic. Arsenic removal employs naturally selected hyperaccumulator ferns, which accumulate very high concentrations of arsenic specifically in above-ground tissues. Elegant two-gene transgenic approaches have been designed for the development of mercury or arsenic phytoremediation technologies. In a plant that naturally hyperaccumulates zinc in leaves, approximately ten key metal homeostasis genes are expressed at very high levels. This outlines the extent of change in gene activities needed in the engineering of transgenic plants for soil clean-up. Further analysis and discovery of genes for phytoremediation will benefit from the recent development of segregating populations for a genetic analysis of naturally selected metal hyperaccumulation in plants, and from comprehensive ionomics data--multi-element concentration profiles from a large number of Arabidopsis mutants. PMID- 15831378 TI - Tuning the cell-cycle engine for improved plant performance. AB - Cell-cycle regulation plays a crucial role in organogenesis, morphogenesis, growth and differentiation and conceptually offers a means to design a next generation of crop plants that outperform traditionally bred ones. However, cell cycle regulation involves a large, highly redundant, set of genes, which complicates unravelling of function in the context of a higher plant. Nevertheless, ten years of molecular cell-cycle research, primarily in the model plant Arabidopsis, have demonstrated its potential for altering plant development. PMID- 15831379 TI - Yield enhancement genes: seeds for growth. AB - Yield is a multifactorial trait, integrating various developmental and physiological processes. Despite this complexity, evidence is mounting that yield can be increased by the genetic modification of single genes. Positive results have been obtained by targeting different yield constituents, indicating that there is ample room for further yield improvement by genetic means. Successful targets include photosynthesis, starch biosynthesis, plant architecture and transcriptional networks controlling plant development. Most of the current data have been obtained in a (semi-)controlled environment and relate to yield calculated on a per plant basis. Demonstrating the ability to transfer these effects to field-grown plants and with reference to yield on a per area unit basis will be a crucial step in establishing the agronomic importance of these findings. PMID- 15831380 TI - Vernalization and flowering time. AB - Vernalization is the process by which flowering is promoted by prolonged exposure to the cold of a typical winter. In certain plant species, the role of vernalization is to suppress the expression of genes that encode repressors of flowering. In Arabidopsis, this suppression is an epigenetic phenomenon in the sense that it is mitotically stable in the spring after the inducing signal, cold, is no longer perceived. This epigenetic silencing results from the modification of the chromatin of flowering repressors. PMID- 15831381 TI - Biotechnology and the domestication of forest trees. AB - Wood is one of the major renewable materials. To compensate for the ever increasing demand for wood and to reduce pressure on native forests, more wood of higher quality will need to be produced on less land by planting highly productive trees. Biotechnology has shown great promise for forest tree improvement and over the past 10 years this field has flourished. Not only has the potential of transgenic trees with optimized yield and quality traits been demonstrated in field trials, but progress in genetical genomics and association genetics promise quantum leaps forward for tree improvement. PMID- 15831382 TI - Sowing the seeds of success: pharmaceutical proteins from plants. AB - Among the many plant-based production systems that have been developed for pharmaceutical proteins, seeds have the useful advantage of accumulating proteins in a relatively small volume and in a stable environment in which they are protected from degradation. Several seed crops, including cereals, grain legumes and oilseeds, have been explored as production platforms, and the first commercial products -- all technical proteins and enzymes -- have already reached the market. Recent studies have explored the use of seeds for the production of pharmaceutical proteins, particularly replacement human proteins, recombinant antibodies and (oral) vaccines. PMID- 15831383 TI - Integrating genomics and metabolomics for engineering plant metabolic pathways. AB - Plant metabolites are characterized by an enormous chemical diversity, every plant having its own complex set of metabolites. This variety poses analytical challenges, both for profiling multiple metabolites in parallel and for the quantitative analysis of selected metabolites. We are only just starting to understand the roles of these metabolites, many of them being involved in adaptations to specific ecological niches and some finding beneficial use (e.g. as pharmaceuticals). Spectacular advances in plant metabolomics offer new possibilities, together with the aid of systems biology, to explore the extraordinary complexity of the plant biochemical capacity. State-of-the art genomics tools can be combined with metabolic profiling to identify key genes that could be engineered for the production of improved crop plants. PMID- 15831384 TI - Prospects in diatom research. AB - Diatoms are unicellular photosynthetic eukaryotes that play a major role in the global cycling of carbon and silicon. They are believed to have arisen from a secondary endosymbiotic event between two eukaryotes, a red alga and a flagellated heterotroph. Recent analysis of a diatom genome indeed reveals a 'mosaic' nature, with genes derived from plant, animal and bacterial lineages. Advances in molecular genomics are facilitating the use of diatom-specific genes or pathways for biotechnology. Another interest is in understanding the artistry of the amorphous silica shell and the underlying biomineralization process. Materials scientists and chemists are now exploiting diatoms to develop new biomimetic approaches and to create silicon-based microdevices with specific features. PMID- 15831386 TI - Functional ingredient production: application of global metabolic models. AB - The biotechnology industry continuously explores new ways to improve the performance of microbial strains in fermentation processes. Recent focus has been on new genome-wide modelling approaches in functional genomics, which aim to take full advantage of genome sequence data, transcription profiling, proteomics and metabolite profiling for strain improvement. The integration of global metabolic models with genetic and regulatory models will be essential for the practice of metabolic engineering for strain improvement to move forward, simply because we cannot rely on our intuition to grasp the complexity of the biological systems involved. PMID- 15831387 TI - Fermented functional foods based on probiotics and their biogenic metabolites. AB - The claimed health benefits of fermented functional foods are expressed either directly through the interaction of ingested live microorganisms, bacteria or yeast with the host (probiotic effect) or indirectly as a result of ingestion of microbial metabolites produced during the fermentation process (biogenic effect). Although still far from fully understood, several probiotic mechanisms of action have been proposed, including competitive exclusion, competition for nutrients and/or stimulation of an immune response. The biogenic properties of fermented functional foods result from the microbial production of bioactive metabolites such as certain vitamins, bioactive peptides, organic acids or fatty acids during fermentation. PMID- 15831388 TI - Probiotic and other functional microbes: from markets to mechanisms. AB - Insight into the diversity and function of the human intestinal microbiota has been stimulated by clinical studies with bacteria that exhibit specific functions and which are marketed as probiotics to positively affect our health. Initial efforts concentrated on establishing sound scientific support for the efficacy of these probiotic bacteria, which mainly include Lactobacillus and Bifidobacterium species. Following these evidence-based functional approaches, considerable research is now focused on the mechanisms of action of probiotic bacteria. The mechanisms identified to date mainly relate to the stimulation of host defence systems, immune modulation and the competitive exclusion of pathogens. Recent efficacy, molecular and genomics-based studies have also been reported for some probiotic strains that have found their position in the market place. PMID- 15831390 TI - The role of sigmaB in the stress response of Gram-positive bacteria -- targets for food preservation and safety. AB - The alternative sigma factor sigmaB modulates the stress response of several Gram positive bacteria, including Bacillus subtilis and the food-borne human pathogens Bacillus cereus, Listeria monocytogenes and Staphylococcus aureus. In all these bacteria, sigmaB is responsible for the transcription of genes that can confer stress resistance to the vegetative cell. Recent findings indicate that sigmaB also plays an important role in antibiotic resistance, pathogenesis and cellular differentiation processes such as biofilm formation and sporulation. Although there are important differences in the regulation of sigmaB and in the set of genes regulated by sigmaB in B. subtilis, B. cereus, L. monocytogenes and S. aureus, there are also some conserved themes. A mechanistic understanding of the sigmaB activation processes and assessment of its regulon could provide tools for pathogen control and inactivation both in the food industry and clinical settings. PMID- 15831389 TI - Prebiotics and other microbial substrates for gut functionality. AB - The intestinal microbiota is of great importance to our health and wellbeing. Modulation of the intestinal microbiota by exogenous and endogenous substrates can be expected to improve various physiological functions of our body, not just those in the intestine. Recently, new targets such as immune function and areas outside the colon have been considered to be influenced by the intestinal microbiota. Novel approaches might include the application of prebiotics in different combinations or the provision of nutrients to different bacterial groups and to different parts of the intestine. PMID- 15831391 TI - Stress tolerance in fungi -- to kill a spoilage yeast. AB - The fungal spoilage of ingredients of food manufacture is an economic problem, often causes product loss and may constitute a health hazard. To effectively combat fungal food spoilage, a mechanistic understanding of tolerance for, and adaptation to, the preservation method used is crucial. Both are dependent on the genetic make-up and growth history of the organism. In the post-genomic era we are arriving at a situation in which, in the model organism Saccharomyces cerevisiae, physiological data, classical molecular biology and whole-genome responses can be combined to obtain explanatory and predictive models for growth. For food spoilage fungi we have not yet reached such a level of understanding, but we may use the knowledge gained for S. cerevisiae for the prevention of spoilage. PMID- 15831392 TI - Exploring fungal biodiversity for the production of water-soluble pigments as potential natural food colorants. AB - The production of many currently authorized natural food colorants has a number of disadvantages, including a dependence on the supply of raw materials and variations in pigment extraction. Fungi provide a readily available alternative source of naturally derived food colorants that could easily be produced in high yields. The recent authorization of a fungal food colorant has fuelled research to explore the extraordinary chemical diversity and biodiversity of fungi for the biotechnological production of pigments as natural food colorants. These studies require an appropriate use of chemotaxonomic tools and a priori knowledge of fungal metabolites to carry out intelligent screening for known or novel colorants as lead compounds. Such screening would result in the preselection of some potential pigment producers and the deselection of pathogenic strains and toxin producers. With advances in gene technology, in the future it should be possible to employ metabolic engineering to create microbial cell factories for the production of food colorants. PMID- 15831393 TI - Cognitive neuroscience. PMID- 15831394 TI - From monkeys to humans: what do we now know about brain homologies? AB - Different primate species, including humans, have evolved by a repeated branching of lineages, some of which have become extinct. The problem of determining the relationships among cortical areas within the brains of the surviving branches (e.g. humans, macaque monkeys, owl monkeys) is difficult for several reasons. First, evolutionary intermediates are missing, second, measurement techniques are different in different primate species, third, species differ in body size, and fourth, brain areas can duplicate, fuse, or reorganize between and within lineages. PMID- 15831395 TI - See me, hear me, touch me: multisensory integration in lateral occipital-temporal cortex. AB - Our understanding of multisensory integration has advanced because of recent functional neuroimaging studies of three areas in human lateral occipito-temporal cortex: superior temporal sulcus, area LO and area MT (V5). Superior temporal sulcus is activated strongly in response to meaningful auditory and visual stimuli, but responses to tactile stimuli have not been well studied. Area LO shows strong activation in response to both visual and tactile shape information, but not to auditory representations of objects. Area MT, an important region for processing visual motion, also shows weak activation in response to tactile motion, and a signal that drops below resting baseline in response to auditory motion. Within superior temporal sulcus, a patchy organization of regions is activated in response to auditory, visual and multisensory stimuli. This organization appears similar to that observed in polysensory areas in macaque superior temporal sulcus, suggesting that it is an anatomical substrate for multisensory integration. A patchy organization might also be a neural mechanism for integrating disparate representations within individual sensory modalities, such as representations of visual form and visual motion. PMID- 15831396 TI - Perceptual learning: specificity versus generalization. AB - Perceptual learning improves performance on many tasks, from orientation discrimination to the identification of faces. Although conventional wisdom considered sensory cortices as hard-wired, the specificity of improvement achieved through perceptual learning indicates an involvement of early sensory cortices. These cortices might be more plastic than previously assumed, and both sum-potential and single cell recordings indeed demonstrate plasticity of neuronal responses of these sensory cortices. However, for learning to be optimally useful, it must generalize to other tasks. Further research on perceptual learning should therefore, in my opinion, investigate first, the conditions for generalization of training-induced improvement, second, its use for teaching and rehabilitation, and third, its dependence on pharmacological agents. PMID- 15831397 TI - Reorganization and plasticity in the adult brain during learning of motor skills. AB - On the basis of brain imaging studies, Doyon and Ungerleider recently proposed a model describing the cerebral plasticity that occurs in both cortico-striatal and cortico-cerebellar systems of the adult brain during learning of new motor skilled behaviors. This theoretical framework makes several testable predictions with regards to the contribution of these neural systems based on the phase (fast, slow, consolidation, automatization, and retention) and nature of the motor learning processes (motor sequence versus motor adaptation) acquired through repeated practice. There has been recent behavioral, lesion and additional neuroimaging studies that have addressed the assumptions made in this theory that will help in the revision of this model. PMID- 15831398 TI - The primate hippocampus: ontogeny, early insult and memory. AB - Recent evidence suggests that in primates, as in rodents, the hippocampus shows a developmental continuum that affects memory abilities from infancy to adulthood. In primates relatively few hippocampal-dependent abilities (e.g. some aspects of recognition memory) are present in early infancy, whereas others (e.g. relational memory) begin to show adult-like characteristics around 2 years of age in monkeys and 5-7 years in humans. Profound and persistent memory loss resulting from insult to the hippocampus in infancy becomes evident in everyday behavior only later in childhood. This pattern of results suggests a maturational gradient within the medial temporal lobe memory system, with most abilities crucially dependent upon the hippocampus emerging in later stages of development, supporting a model of hierarchical organization of memory within the medial temporal lobe. PMID- 15831399 TI - Directing the mind's eye: prefrontal, inferior and medial temporal mechanisms for visual working memory. AB - Human and nonhuman primates have a remarkable ability to recall, maintain and manipulate visual images in the absence of external sensory stimulation. Evidence from lesion, single-unit neurophysiological and neuroimaging studies shows that these visual working memory processes are consistently associated with sustained activity in object-selective inferior temporal neurons. Furthermore, results from these studies suggest that mnemonic activity in the inferior temporal cortex is, in turn, supported by top-down inputs from multimodal regions in prefrontal and medial temporal cortex, and under some circumstances, from the hippocampus. PMID- 15831400 TI - The temporal flexibility of attentional selection in the visual cortex. AB - Visual attention operates by biasing competitive interactions between neural representations, favoring relevant over irrelevant visual inputs. Attention can enhance the processing of relevant information using location-based, feature based or object-based selection mechanisms. Studies using event-related potential and event-related magnetic field recordings, together with functional magnetic resonance imaging, show that the temporal sequencing of these different selection mechanisms is flexible. Depending on the specific processing demands of the experimental task, location-based, feature-based or object-based selection might be given temporal priority on a time scale of tens of milliseconds. PMID- 15831401 TI - To what extent are emotional visual stimuli processed without attention and awareness? AB - In the past few years, important contributions have been made to the study of emotional visual perception. Researchers have reported responses to emotional stimuli in the human amygdala under some unattended conditions (i.e. conditions in which the focus of attention was diverted away from the stimuli due to task instructions), during visual masking and during binocular suppression. Taken together, these results reveal the relative degree of autonomy of emotional processing. At the same time, however, important limitations to the notion of complete automaticity have been revealed. Effects of task context and attention have been shown, as well as large inter-subject differences in sensitivity to the detection of masked fearful faces (whereby briefly presented, target fearful faces are immediately followed by a neutral face that 'masks' the initial face). A better understanding of the neural basis of emotional perception and how it relates to visual attention and awareness is likely to require further refinement of the concepts of automaticity and awareness. PMID- 15831402 TI - Auditory processing--speech, space and auditory objects. AB - There have been recent developments in our understanding of the auditory neuroscience of non-human primates that, to a certain extent, can be integrated with findings from human functional neuroimaging studies. This framework can be used to consider the cortical basis of complex sound processing in humans, including implications for speech perception, spatial auditory processing and auditory scene segregation. PMID- 15831403 TI - The neural basis of first and second language processing. AB - Fundamental breakthroughs in the neurosciences, combined with technical innovations for measuring brain activity, are shedding new light on the neural basis of second language (L2) processing, and on its relationship to native language processing (L1). The long-held assumption that L1 and L2 are necessarily represented in different brain regions in bilinguals has not been confirmed. On the contrary, the available evidence indicates that L1 and L2 are processed by the same neural devices. The neural differences in L1 and L2 representations are only related to the specific computational demands, which vary according to the age of acquisition, the degree of mastery and the level of exposure to each language. Finally, the acquisition of L2 could be considered as a dynamic process, requiring additional neural resources in specific circumstances. PMID- 15831404 TI - Neural substrates of processing syntax and semantics in music. AB - Growing evidence indicates that syntax and semantics are basic aspects of music. After the onset of a chord, initial music-syntactic processing can be observed at about 150-400 ms and processing of musical semantics at about 300-500 ms. Processing of musical syntax activates inferior frontolateral cortex, ventrolateral premotor cortex and presumably the anterior part of the superior temporal gyrus. These brain structures have been implicated in sequencing of complex auditory information, identification of structural relationships, and serial prediction. Processing of musical semantics appears to activate posterior temporal regions. The processes and brain structures involved in the perception of syntax and semantics in music have considerable overlap with those involved in language perception, underlining intimate links between music and language in the human brain. PMID- 15831405 TI - Human motor cortex excitability during the perception of others' action. AB - Neuroscience research during the past ten years has fundamentally changed the traditional view of the motor system. In monkeys, the finding that premotor neurons also discharge during visual stimulation (visuomotor neurons) raises new hypotheses about the putative role played by motor representations in perceptual functions. Among visuomotor neurons, mirror neurons might be involved in understanding the actions of others and might, therefore, be crucial in interindividual communication. Functional brain imaging studies enabled us to localize the human mirror system, but the demonstration that the motor cortex dynamically replicates the observed actions, as if they were executed by the observer, can only be given by fast and focal measurements of cortical activity. Transcranial magnetic stimulation enables us to instantaneously estimate corticospinal excitability, and has been used to study the human mirror system at work during the perception of actions performed by other individuals. In the past ten years several TMS experiments have been performed investigating the involvement of motor system during others' action observation. Results suggest that when we observe another individual acting we strongly 'resonate' with his or her action. In other words, our motor system simulates underthreshold the observed action in a strictly congruent fashion. The involved muscles are the same as those used in the observed action and their activation is temporally strictly coupled with the dynamics of the observed action. PMID- 15831406 TI - The frontal lobes and the regulation of mental activity. AB - Results of neuroimaging and neuropsychological studies of frontal lobe function have been interpreted by some as evidence for specialized modules that are localized to distinct regions of frontal cortex, and that differ in both content and process from those in neighboring regions. These descriptions stand in stark contrast to the many domain-general theoretical accounts of the regulatory role of the frontal lobes in cognition. Recent attempts to understand how general regulatory mechanisms might operate across multiple domains (e.g. working memory, sentence comprehension) have been increasingly important in our understanding of the frontal lobes. PMID- 15831407 TI - Why the frontal cortex in autism might be talking only to itself: local over connectivity but long-distance disconnection. AB - Although it has long been thought that frontal lobe abnormality must play an important part in generating the severe impairment in higher-order social, emotional and cognitive functions in autism, only recently have studies identified developmentally early frontal lobe defects. At the microscopic level, neuroinflammatory reactions involving glial activation, migration defects and excess cerebral neurogenesis and/or defective apoptosis might generate frontal neural pathology early in development. It is hypothesized that these abnormal processes cause malformation and thus malfunction of frontal minicolumn microcircuitry. It is suggested that connectivity within frontal lobe is excessive, disorganized and inadequately selective, whereas connectivity between frontal cortex and other systems is poorly synchronized, weakly responsive and information impoverished. Increased local but reduced long-distance cortical cortical reciprocal activity and coupling would impair the fundamental frontal function of integrating information from widespread and diverse systems and providing complex context-rich feedback, guidance and control to lower-level systems. PMID- 15831408 TI - Reading and reading disturbance. AB - Recent functional neuroimaging studies are generating novel insights into our knowledge of skilled and disturbed reading. In neurologically normal subjects, a double dissociation in neural activation in response to reading words and pseudowords has been revealed that corresponds to that observed in the comparison of semantic and phonological tasks. In patients with acquired dyslexia, functional imaging is demonstrating re-organisation within the reading system; in developmental dyslexia, functional imaging is being used to identify the impact of rehabilitation. Together, these findings have implications for cognitive models of reading that have previously relied on input from behavioural data. PMID- 15831409 TI - Changes in cerebral functional organization during cognitive development. AB - It has been just under a decade since contemporary neuroimaging tools, such as functional magnetic resonance imaging, were first applied to developmental questions. These tools provide invaluable information on how brain anatomy, function and connectivity change during development. Studies using these methods with children and adolescents show that brain regions that support motor and sensory function mature earliest, whereas higher-order association areas, such as the prefrontal cortex, which integrate these functions, mature later. PMID- 15831410 TI - Brain aging: reorganizing discoveries about the aging mind. AB - New discoveries challenge the long-held view that aging is characterized by progressive loss and decline. Evidence for functional reorganization, compensation and effective interventions holds promise for a more optimistic view of neurocognitive status in later life. Complexities associated with assigning function to age-specific activation patterns must be considered relative to performance and in light of pathological aging. New biological and genetic markers, coupled with advances in imaging technologies, are enabling more precise characterization of healthy aging. This interdisciplinary, cognitive neuroscience approach reveals dynamic and optimizing processes in aging that might be harnessed to foster the successful aging of the mind. PMID- 15831412 TI - Meta-analysis and structured literature review in radiology. AB - The overall goal of a systematic review or meta-analysis is to combine results of previous studies to arrive at summary conclusions about a body of research. In radiology, systematic reviews or meta-analyses can be used to calculate a summary estimate of effect size of a treatment that used imaging data to assess outcomes in observational or randomized controlled clinical trials, estimate the clinical effectiveness of an imaging-guided therapy procedure, evaluate the summary diagnostic accuracy of an imaging test, or synthesize results of economic evaluations that used imaging data. This article outlines the general concepts of structured literature reviews and discusses the approaches for conducting a meta analysis in radiology, emphasizing the methods available for data synthesis and handling heterogeneity between and among studies. PMID- 15831413 TI - Improving outcomes in radiology: bringing computer-based decision support and education to the point of care. AB - Many computer applications have been developed in radiology and other medical disciplines to help physicians make decisions. Artificial intelligence (AI)--an approach to computer-based manipulation of symbols to simulate human reasoning- forms the basis of many of these systems. This article's goals are to: acquaint the reader with the motivations and opportunities for computer-based medical decision support systems; identify AI techniques and applications in radiology decision making; assess the impact of these technologies; and consider new directions and opportunities for AI in radiology. Among the exciting new directions is the use of AI to integrate radiology reporting, online decision support, and just-in-time learning to provide useful information and continuing education that is embedded within a radiologist's daily workflow. PMID- 15831414 TI - Development of radiology prediction models using feature analysis. AB - RATIONALE AND OBJECTIVES: This article provides an introduction to prediction models and their application in diagnostic imaging research. Prediction models capitalize on the different degrees of association among variables to make a prediction of a health state, formulate a rule, or quantify individual contributions of various predictor variables. The purpose of this article is to elucidate the rationale, implication, and interpretation of prediction models using imaging features. MATERIALS AND METHODS: The techniques and challenges of developing, testing, and implementing prediction models are described. Prediction model development methods are similar to data-mining techniques. RESULTS: Learning objectives are to review prediction rule (model) methods, learn how prediction models may be applied to feature analysis, and understand the challenges of developing, testing, and implementing prediction models. PMID- 15831415 TI - Bayesian networks: computer-assisted diagnosis support in radiology. AB - Medical knowledge is growing at an explosive rate. While the availability of pertinent data has the potential to make the task of diagnosis more accurate, it is also increasingly overwhelming for physicians to assimilate. Using artificial intelligence techniques, a computer can process large amounts of data to help physicians manage the growing body of medical knowledge and thereby make better decisions. Computer-assisted diagnosis support is of particular interest to the diagnostic imaging community because radiologists must integrate huge amounts of data in order to diagnose disease. Bayesian networks, among the most promising artificial intelligence techniques available, enable computers to store knowledge and estimate the probability of outcomes based on probability theory. The article describes what a Bayesian network is and how it works using a system in mammography for illustration. A comparison of Bayesian networks with other types of artificial intelligence methods, specifically neural networks and case-based reasoning, clarifies the unique features and the potential of these systems to aid radiologists in the decisions they make every day. PMID- 15831416 TI - The socioeconomic aspects of information technology for health care with emphasis on radiology. AB - RATIONALE AND OBJECTIVES: Information technology is the key to cost effective and error free medical care in the United States and the only problem is that there is not enough of it yet. During the past 15 years, billions of dollars have been spent on information technology for health care with very little benefit but significant adverse effects on patients, physicians, and nurses. The truth about health care information technology (HIT) probably lies somewhere between these extreme statements, representing technophile and skeptical views, respectively. MATERIALS AND METHODS: There is no doubt that computer and communication hardware has reached a state of sophistication and availability in which any and all necessary information can be generated, stored, and distributed to health care workers in support of their patient care tasks. The barriers to rapid and widespread development and diffusion of cost effective and practically useful HIT are exclusively related to human factors. RESULTS: This article explores some of the organizational, cultural, cognitive, and economic forces that interact to influence success of HIT initiatives in health care organizations. A key point to be recognized is that the intrinsically handcrafted nature of health care work combined with high degrees of complexity and contingency make it impossible to "computerize" with the same ease and completeness of other industries. The major thrust of the argument is that designers of information systems and health care informatics managers must meet needs of patients and care providers. The software they create and implement should promote, support, and enhance the existing processes of health care rather than seeking to dictate how direct care providers should do their work. CONCLUSIONS: Instead of looking for "buy in" from physicians and nurses, the informatics community must return the authority over functional specification of patient care information systems to them--where it belonged in the first place. This same lesson about computer technology and organizational politics is also being learned in the business community, where executives are reclaiming responsibility for mission critical informatics decisions. PMID- 15831417 TI - Three-dimensional assessment of MR imaging-guided percutaneous cryotherapy using multi-performer repeated segmentations: the value of supervised learning. AB - RATIONALE AND OBJECTIVES: Accurate and reproducible segmentations of two dimensional images are an important prerequisite for assessing tumor ablations three dimensionally (3D). We evaluated whether supervised learning methods would improve multiperformer repeated segmentations of magnetic resonance images (MRI) obtained before and after MRI-guided cryotherapy of renal cell carcinoma. MATERIALS AND METHODS: Three medical students independently performed five manual segmentations of a biopsy-proven renal cell carcinoma that was treated with percutaneous MRI-guided cryotherapy. Using pretreatment (T2-weighted fast recovery fast spin echo [FRFSE]) and posttreatment (T1-weighted, fat-suppressed, dynamically enhanced) MRIs, regions of tumor cryonecrosis were segmented. The same tasks were repeated after an experienced abdominal radiologist provided supervised learning. Segmentation sensitivity was compared with an estimated 3D ground truth via voxel counts for regions of tumor, both before and after treatment, and for the regions of cryonecrosis. The sensitivity of each repeated segmentation was compared against the estimated ground truth using sensitivity, overlap index, and volume (mL). RESULTS: Supervised learning significantly improved posttreatment segmentation sensitivity (P = .03). With supervised learning, the ranges of the performance metrics over the segmentation performers were: pretreated tumor, sensitivity 0.902-0.999, overlap index 0.935-0.961, and volume 19.15-23.71 mL; posttreated tumor, sensitivity 0.923-0.991, overlap index 0.952-0.981, and volume 20.67-22.70 mL; in the ablation zone, sensitivity 0.938 0.969, overlap index 0.940-0.962, and volume 31.79-32.36 mL. CONCLUSIONS: Supervised learning improved multiperformer repeated segmentations of MRIs obtained before and after MRI-guided percutaneous cryotherapy of renal cell carcinoma. These methods may prove useful in aiding the 3D assessment of percutaneous tumor ablations. PMID- 15831418 TI - Associations in breast and colon cancer screening behavior in women. AB - RATIONALE AND OBJECTIVES: Gender-based psychosocial factors appear to influence colorectal cancer (CRC) screening adherence. Given its near-universal acceptance by the public, screening mammography represents a potential "teachable moment" for educating patients about the risk of CRC. Accordingly, to better understand screening behaviors among women, data from the Behavioral Risk Factors Surveillance Survey (BRFSS) were analyzed to identify potential relationships that would allow interventions to enhance CRC screening. MATERIALS AND METHODS: Women 50 years and older who participated in the BRFSS 2001 survey were included in the analysis. Colorectal, breast, and cervical cancer screening adherence with American Cancer Society guidelines was determined. We identified the association between breast and cervical cancer screening adherence and general health and demographic characteristics with CRC screening adherence. RESULTS: After adjustment for sociodemographic factors in a multivariate analysis, women 60-69 years old (adjusted odds ratio [OR], 1.50; P < .01) and 70-79 years old (adjusted OR, 1.39; P < .01), having achieved at least some high school (adjusted OR, 1.62; P < .01) or college (adjusted OR, 2.11; P < .01) education, having health coverage (adjusted OR, 1.67; P < .01) or a personal physician (adjusted OR, 1.60; P < .01), and adherence to screening mammography (adjusted OR, 2.42; P < .01) and Pap smear (adjusted OR, 1.70; P < .01) were independently associated with an increased likelihood CRC screening adherence. Women in self-reported good general health were less likely to have adhered to CRC screening guidelines (adjusted OR, 0.79; P < .01). Current smokers were also less likely to have adhered to CRC screening guidelines than were women who never smoked or formerly smoked (adjusted OR, 0.76; P < .01). Participants who adhered to both mammography and Pap smear guidelines were significantly more likely to adhere to CRC screening (51.5% CRC screening adherence) compared with women who adhered to neither screening test (8.2% CRC screening adherence), with an adjusted OR of 5.67 (P < .001). Participants who adhered to both mammography and Pap smear guidelines were significantly more likely to adhere to CRC screening than were women who adhered to either screening test (38.0% CRC screening adherence) with an adjusted OR of 1.94 (P < .001). CONCLUSION: Women with up-to-date mammography and cervical cancer screening were more likely to be up-to-date with CRC screening. Regardless of the increased association between non-CRC-related cancer screening and CRC screening, rates of CRC screening utilization remained low in these otherwise compliant populations. PMID- 15831419 TI - Statistical validation of brain tumor shape approximation via spherical harmonics for image-guided neurosurgery. AB - RATIONALE AND OBJECTIVES: Surgical planning now routinely uses both two dimensional (2D) and three-dimensional (3D) models that integrate data from multiple imaging modalities, each highlighting one or more aspects of morphology or function. We performed a preliminary evaluation of the use of spherical harmonics (SH) in approximating the 3D shape and estimating the volume of brain tumors of varying characteristics. MATERIALS AND METHODS: Magnetic resonance (MR) images from five patients with brain tumors were selected randomly from our MR guided neurosurgical practice. Standardized mean square reconstruction errors (SMSRE) by tumor volume were measured. Validation metrics for comparing performances of the SH method against segmented contours (SC) were the dice similarity coefficient (DSC) and standardized Euclidean distance (SED) measure. RESULTS: Tumor volume range was 22,413-85,189 mm3, and range of number of vertices in triangulated models was 3674-6544. At SH approximations with degree of at least 30, SMSRE were within 1.66 x 10(-5) mm(-1). Summary measures yielded a DSC range of 0.89-0.99 (pooled median, 0.97 and significantly >0.7; P < .001) and an SED range of 0.0002-0.0028 (pooled median, 0.0005). CONCLUSION: 3D shapes of tumors may be approximated by using SH for neurosurgical applications. PMID- 15831420 TI - Assessing patient utilities for varying degrees of low back pain. AB - RATIONALE AND OBJECTIVES: We sought to quantify patient preferences for mild, moderate, and severe low back pain via time-tradeoff analysis and utility measurement. MATERIALS AND METHODS: Forty-one patients being treated for low back pain in a tertiary care teaching hospital participated in the study. Patients were asked to decide which of three health states they were currently experiencing as well as which of the three health states was the worst experienced during their lifetime. A time-tradeoff analysis was performed, during which patients were asked the amount of time in each of the health states they would exchange for complete resolution of symptoms. We correlated (1) subjects' current health state with reported utility and (2) degree of previous low back pain with results of time-tradeoff measurements. RESULTS: All patients were willing to trade a greater number of life-years for resolution of symptoms given a more severe perceived health state. Utility decreased as severity of back pain scenarios increased, with an average utility of 0.93 +/- 0.11 for mild, 0.65 +/- 0.21 for moderate, and 0.18 +/- 0.17 for severe pain. No significant difference in time-tradeoff among subjects was identified based upon current health state on the day of interview. A statistically significant difference was seen in patients' willingness to trade time among those who had actually experienced severe pain versus those who had not. Kendall's correlation revealed that subjects who had experienced severe back pain exhibited significantly lower utilities (P < 0.01) compared with subjects who had never experienced severe pain. CONCLUSIONS: As expected, patients with severe low back pain were willing to sacrifice more potential years of life for resolution of symptoms, suggesting time-tradeoff can accurately reflect patient utility. 2. However, we found no correlation between a subject's current health state and reported utility. PMID- 15831421 TI - Value added radiology: using existing radiology services to improve cancer screening. PMID- 15831422 TI - Support vector machines committee classification method for computer-aided polyp detection in CT colonography. AB - RATIONALE AND OBJECTIVES: A new classification scheme for the computer-aided detection of colonic polyps in computed tomographic colonography is proposed. MATERIALS AND METHODS: The scheme involves an ensemble of support vector machines (SVMs) for classification, a smoothed leave-one-out (SLOO) cross-validation method for obtaining error estimates, and use of a bootstrap aggregation method for training and model selection. Our use of an ensemble of SVM classifiers with bagging (bootstrap aggregation), built on different feature subsets, is intended to improve classification performance compared with single SVMs and reduce the number of false-positive detections. The bootstrap-based model-selection technique is used for tuning SVM parameters. In our first experiment, two independent data sets were used: the first, for feature and model selection, and the second, for testing to evaluate the generalizability of our model. In the second experiment, the test set that contained higher resolution data was used for training and testing (using the SLOO method) to compare SVM committee and single SVM performance. RESULTS: The overall sensitivity on independent test set was 75%, with 1.5 false-positive detections/study, compared with 76%-78% sensitivity and 4.5 false-positive detections/study estimated using the SLOO method on the training set. The sensitivity of the SVM ensemble retrained on the former test set estimated using the SLOO method was 81%, which is 7%-10% greater than the sensitivity of a single SVM. The number of false-positive detections per study was 2.6, a 1.5 times reduction compared with a single SVM. CONCLUSION: Training an SVM ensemble on one data set and testing it on the independent data has shown that the SVM committee classification method has good generalizability and achieves high sensitivity and a low false-positive rate. The model selection and improved error estimation method are effective for computer-aided polyp detection. PMID- 15831423 TI - Comparative analysis of logistic regression and artificial neural network for computer-aided diagnosis of breast masses. AB - RATIONALE AND OBJECTIVE: To compare logistic regression and artificial neural network for computer-aided diagnosis on breast sonograms. MATERIALS AND METHODS: Ultrasound images of 24 malignant and 30 benign masses were analyzed quantitatively for margin sharpness, margin echogenicity, and angular variation in margin. These features and age of patients were used with two pattern classifiers, logistic regression, and an artificial neural network to differentiate between malignant and benign masses. The performance of two methods was compared by receiver operating characteristic (ROC) analysis. RESULTS: The area under the ROC curve Az (+/-SD) of the logistic regression analysis was 0.853 +/- 0.059 with 95% confidence limit (0.760-0.950). The area under the ROC curve of the artificial neural network analysis was 0.856 +/- 0.058 with 95% confidence limit (0.734-0.936). Although both the logistic regression and the artificial neural network had the same area under the ROC curve, the shapes of two curves were different. At 95% sensitivity, the artificial neural network had 76.5% specificity, whereas logistic regression had 64.7% specificity. CONCLUSION: There was no difference in performance between logistic regression and the artificial neural network as measured by the area under the ROC curve. However, at a fixed 95% sensitivity, the artificial neural network had higher (12%) specificity compared with logistic regression value. PMID- 15831424 TI - Solitary pulmonary nodule diagnosis on CT: results of an observer study. AB - RATIONALE AND OBJECTIVES: To investigate the performance of observers with different levels of experience in distinguishing between benign and malignant solitary pulmonary nodules (SPN) on CT, and to determine the effects on interpretation of three different conditions: image data alone, the addition of clinical data, and the addition of output from a computer-aided diagnosis (CAD) system. MATERIALS AND METHODS: 28 thin-section CT datasets of SPNs with proven diagnoses (15 malignant and 13 benign) were used to measure observer performance. Readers were categorized according to their experience and read the cases in random order. For each case readers were asked to assign a level of confidence on a scale from 0.0-1.0 (0.0 benign, 1.0 malignant) for the diagnosis of the nodule. Each reader scored the cases based on review of image data alone (phase 1), then with limited clinical data (phase 2), and finally with CAD output (phase 3). To assess performance, multiple reader multiple case (MRMC) receiver operating characteristic (ROC) analysis was used. RESULTS: 2 thoracic radiologists, 1 thoracic radiology fellow, 2 nonthoracic radiologists, and 3 radiology residents read the cases. The average area under the ROC curve for all readers (A(z)) at each stage was 0.68, 0.75, and 0.81, for image data alone, with clinical data, and with CAD output respectively. The difference in performance between phases (2 and 3) and (1 and 3) was significantly different (P = 0.018 and P = 0.020). However, the difference between phases (1 and 2) was not significantly different (P = 0.155). CONCLUSION: Diagnostic performance increased significantly with the addition of CAD output. With further validation CAD output may play a significant role in SPN management. PMID- 15831425 TI - Comparison of iron oxide labeling properties of hematopoietic progenitor cells from umbilical cord blood and from peripheral blood for subsequent in vivo tracking in a xenotransplant mouse model XXX. AB - RATIONALE AND OBJECTIVES: To compare and optimize ferumoxides labeling of human hematopoietic progenitor cells from umbilical cord blood and from peripheral blood for subsequent in vivo tracking with a clinical 1.5 T MR scanner. MATERIALS AND METHODS: Human hematopoietic progenitor cells, derived from umbilical cord blood or peripheral blood, were labeled with Ferumoxides by simple incubation or lipofection. Cellular iron uptake was quantified with spectrometry. Then, 3 x 10(7)-labeled cells were injected into the tail vein of 12 female nude Balb/c mice. The mice underwent magnetic resonance imaging before and 24 hours after injection. Precontrast and postcontrast signal intensities of liver, spleen, and bone marrow were measured and tested for significant differences with the t-test. Immunostains served as a histopathologic standard of reference. RESULTS: After labeling by simple incubation, only umbilical cord blood cells, but not peripheral blood cells, showed a significant iron uptake and could be tracked in vivo with magnetic resonance imaging. Using lipofection, both cell types could be tracked in vivo. A significant decline in signal intensity was observed in liver, spleen, and bone marrow at 24 hours after injection of efficiently labeled ferumoxides cells (P < .05). Histopathology proved the distribution of iron oxide labeled cells to these organs. CONCLUSION: Hematopoietic progenitor cells from umbilical cord blood can be labeled by simple incubation with an Food and Drug Administration-approved magnetic resonance contrast agent with sufficient efficiency to provide an in vivo cell tracking at 1.5 T. Progenitor cells from peripheral blood need to be labeled with adjunctive transfection techniques to be depicted in vivo at 1.5 T. PMID- 15831426 TI - Design, analysis and simulation for development of the first clinical micro-CT scanner. AB - In this article, we propose to develop the first clinical micro-CT (CMCT) system for human temporal bone imaging in vivo. This CMCT system consists of medical CT and micro-CT scanners either as separate components or in a combination, a cross modality registration mechanism such as a facial surface scanner, and associated software. This system integrates the strengths of state-of-the-art medical CT and micro-CT techniques to achieve a spatial resolution that is much higher than currently available for inner ear imaging at acceptable dose levels. Our design, analysis, and simulation results demonstrate that the CMCT system is feasible for inner ear imaging and other clinical applications. For example, the CMCT system has the potential to improve the safety of guiding cochlear implant electrodes within the inner ear and assist the placement of inner ear microcatheters for delivery of gene modification therapy or administration of neurotrophic factors. Imaging of microarchitectures of the cancellous bone would be also an important application. PMID- 15831427 TI - Results from the problem-solving sessions at the 52nd annual meeting of the Association of University Radiologists. PMID- 15831429 TI - Radiation safety. PMID- 15831430 TI - Developing the role of the social worker as coordinator of services at the surrogate parenting center. AB - A law permitting couples to conceive biological children through surrogacy was legislated in Israel in March 1996. The Rambam Medical Center has established the only nonprofit Surrogate Parenting Center at a public hospital in Israel. The multidisciplinary teamwork at the Center is case managed by a social worker. An important role of the social work intervention is consultation and support for the couple and the surrogate at all stages of the process. The case study presented in the article illustrates the need for sensitive and professional intervention due to the complexity of the surrogacy process and the crisis it involves for both the surrogate and the couple. In light of the growing parenting surrogacy cases in the United States, Europe, and Israel, a structured social work intervention model is described, which may be implemented at public or private surrogate parenting centers. PMID- 15831431 TI - An evaluation of a breast-feeding education intervention among Spanish-speaking families. AB - This study assessed the impact of an educational breast-feeding intervention on the knowledge, attitudes and perceived ability to access breast-feeding resources among Spanish-speaking Latino families making breast-feeding decisions. Conducted in cooperation with the local office of The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), this university-based, multidisciplinary outreach and research project was held in a semirural part of Utah, where Latinos constitute the fastest and largest growing group of the population and have the lowest breast-feeding duration rates. The results suggested that teaching a lactation class in Spanish to Latino women significantly increased their willingness to breast-feed and empowered them by increasing their belief that they could breast-feed even if they work or attend school, that they would not have a problem with insufficient milk, and that they would not need to limit their diet to breast-feed. Additionally, the study indicated that Latino women are more likely to access health care services from agencies providing services in Spanish, regardless of their secondary fluency in English. PMID- 15831432 TI - Coexistent concerns: assessing the social and health needs of dental clinic patients. AB - University dental clinic professionals identified increasing numbers of patients with complex problems and generated 2 research questions: (1) Are there significant health and social concerns within the dental clinic population that indicate the need for high-risk screening and social work services?; and (2) How do age, gender, and income influence health and social concerns in this population? This exploratory descriptive cross-sectional study employed a brief self-report survey in a clinic waiting area. Logistic regression was used to understand the influence of age, gender and income on the existence of specific concerns. Results indicate that caregiving, finances and health are issues for 1/3 of the participants and that 44% endorsed 2 or more concerns. These findings were used to develop a routine high-risk screening tool for dental clinic patients and social work services within the clinic; they suggest that community clinics with dental services are an important place for identifying complex unmet needs. PMID- 15831433 TI - The promise of post-menopausal pregnancy (PMP). AB - Postmenopausal pregnancy (PMP) is presented as a means of alleviating women's suffering by helping them realize their wish for a child of their "own." The availability of IVF technology and oocyte donation, both necessary for PMP, create the illusion of unlimited female fertility. The aim of this paper is twofold: (1) to examine the psychosocial, health, and ethical aspects of PMP; and (2) to answer the question of whether PMP's promise of unlimited fertility really offers more personal freedom and control for women. The paper concludes that PMP seems to increase, rather than to decrease, human suffering. PMID- 15831434 TI - Levels of directiveness: a contextual analysis. AB - Practitioners' decisions concerning levels of directiveness, reflecting their efforts to persuade clients, parallel their decisions about autonomy and beneficence. This qualitative study explores the levels of directiveness employed by social workers with the goal of expanding our understanding of the factors related to these practice decisions. An interview guide was used with a convenience sample of seventeen home health care social workers. Through the use of constant comparative analysis, six levels of directiveness and five related factors were found. This study calls our attention to the ethical dimensions of practice decisions and raises questions about the possible influence of practice context on practitioners' use of levels of directiveness. PMID- 15831436 TI - P-glycoprotein-independent apoptosis induction by a novel synthetic compound, MMPT [5-[(4-methylphenyl)methylene]-2-(phenylamino)-4(5H)-thiazolone]. AB - To develop new anticancer agents that are effective for treatment of chemoresistant tumors, we screened a chemical library for compounds that can effectively kill both paclitaxel-sensitive lung cancer cell H460 and P glycoprotein-overexpressing paclitaxel-resistant cell H460/TaxR. A synthetic compound, MMPT (5-[(4-methylphenyl)methylene]-2-(phenylamino)-4(5H)-thiazolone), was identified to induce cytotoxic effects in both H460 and H460/TaxR cells but not in normal fibroblasts. MMPT effectively inhibited the growth of several human lung cancer cell lines in a dose-dependent manner, with 50% inhibitory concentrations ranging from 4.9 to 8.0 microM. The inhibitory effect on cancer cells is independent of the status of p53 and P-glycoprotein. Moreover, MMPT had no obvious toxic effects on normal human fibroblasts and mesenchymal stem cells at the 50% inhibitory concentration for lung cancer cell lines. Treating lung cancer cells with MMPT-induced apoptosis with caspase-3, -8, -9, and poly(ADP ribose) polymerase cleavage and cytochrome c release from mitochondria. MMPT induced apoptosis was abrogated when c-Jun N-terminal kinase (JNK) activation was blocked with a specific JNK inhibitor, SP600125. Furthermore, in vivo administration of MMPT suppressed human H460 xenograft tumor growth in nude mice. Our results suggest that MMPT may induce tumor-selective cell killing in both P glycoprotein-negative and -positive cancer cells and could be a new anticancer agent for treatment of refractory tumors. PMID- 15831437 TI - Modulation of human 5-hydroxytryptamine type 3AB receptors by volatile anesthetics and n-alcohols. AB - Functional 5-hydroxytryptamine type 3 (5-HT3) receptors can be formed by 5-HT3A subunits alone or in combination with the 5-HT3B subunit, but only the 5-HT3A receptor has been previously studied with respect to the modulation by volatile anesthetics and n-alcohols. Using two-electrode voltage-clamp, we show for the first time the modulation of heteromeric human (h)5-HT3AB receptors, expressed in Xenopus oocytes, by a series of n-alcohols and halogenated volatile anesthetics. At twice their anesthetic concentration, compounds having a molecular volume of less than 110 A3 enhanced submaximal 5-HT-evoked current. Compounds larger than 110 A3 inhibited submaximal 5-HT-evoked current. In experiments examining 5-HT concentration-response relationships, chloroform and butanol caused a slight decrease in the 5-HT EC50. Sevoflurane and octanol inhibited 5-HT-evoked current at all 5-HT concentrations tested but had no effect upon the 5-HT EC50. Compared with previous data on homomeric h5-HT3A receptors, the presence of the h5-HT3B subunit reduces the enhancement of h5-HT3 receptors by smaller halogenated volatile anesthetics and n-alcohols. In summary, these results suggest that heteromeric h5-HT3AB receptors are modulated by halogenated volatile anesthetics at clinically relevant concentrations, in addition to n-alcohols, suggesting that these receptors may be another physiological target for these compounds. The modulation is dependent upon the molecular volume of the compound, further supporting the concept of an anesthetic binding pocket of limited volume common on other Cys-loop ligand-gated ion channels. Incorporation of the 5-HT3B subunit alters either the anesthetic binding site or the allosteric interactions between anesthetic binding and channel opening. PMID- 15831438 TI - Effects of alpha-dendrotoxin on K+ currents and action potentials in tetrodotoxin resistant adult rat trigeminal ganglion neurons. AB - To determine whether the alpha-dendrotoxin (alpha-DTX)-sensitive current [D current, slow inactivating transient current (I(D))] contributes to the modification of neuronal function in small-diameter adult rat trigeminal ganglion (TG) neurons insensitive to 1 microM tetrodotoxin (TTX), we performed two different types of experiments. In the voltage-clamp mode, two distinct K+ current components, a fast inactivating transient current (I(A)) and a dominant sustained current (I(K)), were identified. Alpha-DTX (0.1 microM), ranging from 0.001 to 1 microM, maximally decreased I(A) by approximately 20% and I(K) by approximately 16.1% at a +50-mV step pulse, and 0.1 microM alpha-DTX application increased the number of action potentials without changing the resting membrane potential. Irrespective of the absence and presence of 0.1 microM alpha-DTX, applications of 4-aminopyridine (4-AP; 0.5 mM) and tetraethylammonium (TEA; 2 mM) inhibited approximately 50% inhibition of I(A) and I(K), respectively. 4-AP (0.5 mM) depolarized the resting membrane potential and increased the number of action potentials in the absence or presence of 0.1 microM alpha-DTX. TEA prolonged the duration of action potentials in the absence or presence of 0.1 microM alpha-DTX. These results suggest that I(D) contributes to the modification of neuronal function in adult rat TTX-resistant TG neurons, but after the loss of I(D) due to 0.1 microM alpha-DTX application, 4-AP (0.5 mM) and TEA (2 mM) still regulate the intrinsic firing properties of action potential number and shape. PMID- 15831439 TI - Pharmacological characterization of ecstasy synthesis byproducts with recombinant human monoamine transporters. AB - Ecstasy samples often contain byproducts of the illegal, uncontrolled synthesis of N-methyl-3,4-methylenedioxy-amphetamine or 3,4-methylenedioxymethamphetamine (MDMA). MDMA and eight chemically defined byproducts of MDMA synthesis were investigated for their interaction with the primary sites of action of MDMA, namely the human plasmalemmal monamine transporters for norepinephrine, serotonin, and dopamine [(norepinephrine transporter (NET), serotonin transporter (SERT), and dopamine transporter (DAT)]. SK-N-MC neuroblastoma and human embryonic kidney cells stably transfected with the transporter cDNA were used for uptake and release experiments. Two of the eight compounds, 1,3-bis (3,4 methylenedioxyphenyl)-2-propanamine (12) and N-formyl-1,3-bis (3,4 methylenedioxyphenyl)-prop-2-yl-amine (13) had uptake inhibitory potencies with IC50 values in the low micromolar range similar to MDMA. Compounds with nitro instead of amino groups and a phenylethenyl instead of a phenylethyl structure or a formamide or acetamide modification had IC50 values beyond 100 microM. MDMA, 12, and 13 were examined for induction of carrier-mediated release by superfusion of transporter expressing cells preloaded with the metabolically inert transporter substrate [3H]1-methyl-4-phenylpyridinium. MDMA induced release mediated by NET, SERT, or DAT with EC50 values of 0.64, 1.12, and 3.24 microM, respectively. 12 weakly released from NET- and SERT-expressing cells with maximum effects less than one-tenth of that of MDMA and did not release from DAT cells. 13 had no releasing activity. 12 and 13 inhibited release induced by MDMA, and the concentration dependence of this effect correlated with their uptake inhibitory potency at the various transporters. These results do not support a neurotoxic potential of the examined ecstasy synthesis byproducts and provide interesting structure-activity relationships on the transporters. PMID- 15831441 TI - Effects of azumolene on Ca2+ sparks in skeletal muscle fibers. AB - Azumolene is an analog of dantrolene, the only approved medicine for treatment of malignant hyperthermia (MH). The pharmacological mechanism of these drugs is to inhibit skeletal muscle sarcoplasmic reticulum (SR) Ca2+ release by modulating the activity of the SR ryanodine receptor (RyR) Ca2+ release channel. To investigate the effects of azumolene on SR Ca2+ channel gating within skeletal muscle fibers, we monitored Ca2+ sparks in permeabilized frog skeletal muscle fibers. Application of 0.0001 to 10 microM azumolene suppressed the frequency of spontaneous Ca2+ sparks in a dose-dependent manner (EC50 = 0.25 microM; Hill coefficient = 1.44), but it did not cause systematic dose-dependent effects on the properties of the Ca2+ sparks. These results suggest that azumolene decreases the likelihood of Ca2+ release channel openings that initiate Ca2+ sparks, thereby decreasing spark frequency, but it has little effect on aggregate Ca2+ channel open times during a spark. To assess azumolene inhibition of RyRs activated in a manner analogous to those activated during an MH episode, we applied DP4, a synthetic peptide corresponding to a central region of RyR1 (Leu2442 to Pro2477), which mimics an MH modification. Azumolene also decreased Ca2+ spark frequency in a dose-dependent manner without altering spark properties in the DP4 MH model. We conclude that azumolene suppresses the opening rate but not the open time of RyR Ca2+ release channels within skeletal fibers. PMID- 15831440 TI - Rofecoxib produces intestinal but not gastric damage in the presence of a low dose of indomethacin in rats. AB - Indomethacin in small doses is known to inhibit prostaglandin (PG) production, yet it does not damage the gastrointestinal mucosa. We examined whether a cyclooxygenase (COX)-2 inhibitor induces gastrointestinal damage in the presence of a low dose of indomethacin and investigated the ulcerogenic mechanism in relation to COX-2 expression. Rats with or without 18-h fasting were administered rofecoxib (a selective COX-2 inhibitor; 10 or 30 mg/kg p.o.) in the absence or presence of indomethacin (3 mg/kg p.o.), and the gastric or intestinal mucosa was examined 8 and 24 h later, respectively. Neither indomethacin nor rofecoxib alone caused damage in the stomach or small intestine. However, indomethacin damaged the small intestine in the presence of rofecoxib, yet the same treatment did not damage the stomach. Indomethacin reduced the mucosal PGE2 content in both tissues, whereas rofecoxib did not. The COX-2 mRNA was up-regulated in the intestine but not the stomach after indomethacin treatment, and the reduced PGE2 content was significantly recovered later only in the small intestine, in a rofecoxib-inhibitable manner. Indomethacin produced hypermotility in the small intestine but not the stomach, whereas rofecoxib had no effect. These results suggest that the PG deficiency caused by a low dose of indomethacin produces hypermotility and COX-2 expression in the small intestine but not the stomach, resulting in damage when COX-2 is inhibited. It is assumed that the hypermotility response is a key event in the expression of COX-2 and thereby important in the development of mucosal damage in the gastrointestinal tract. PMID- 15831442 TI - Beneficial effects of a new 20-hydroxyeicosatetraenoic acid synthesis inhibitor, TS-011 [N-(3-chloro-4-morpholin-4-yl) phenyl-N'-hydroxyimido formamide], on hemorrhagic and ischemic stroke. AB - The present study characterized the effects of TS-011 [N-(3-chloro-4-morpholin-4 yl) phenyl-N'-hydroxyimido formamide], a new selective inhibitor of the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE), on the metabolism of arachidonic acid by human and rat renal microsomes and the inhibitory effects of this compound on hepatic cytochrome P450 enzymes involved in drug metabolism. The effects of TS-011 on the fall in cerebral blood flow following subarachnoid hemorrhage (SAH) and in reducing infarct size in ischemic stroke models were also examined since 20-HETE may contribute to the development of cerebral vasospasm. TS-011 inhibited the synthesis of 20-HETE by human renal microsomes and recombinant CYP4A11 and 4F2, 4F3A, and 4F3B enzymes with IC50 values around 10 to 50 nM. It had no effect on the activities of CYP1A, 2C9, 2C19, 2D6, or 3A4 enzymes. TS-011 inhibited the synthesis of 20-HETE by rat renal microsomes with an IC50 of 9.19 nM, and it had no effect on epoxygenase activity at a concentration of 100 microM. TS-011 (0.01-1 mg/kg i.v.) reversed the fall in cerebral blood flow and the increase in 20-HETE levels in the cerebrospinal fluid of rats after SAH. TS-011 also reduced the infarct volume by 35% following transient ischemic stroke and in intracerebral hemorrhage in rats. Injection of 20-HETE (8 or 12 mg/kg) into the carotid artery produced an infarct similar to that seen in the ischemic stroke model. These studies indicate that blockade of the synthesis of 20-HETE with TS-011 opposes cerebral vasospasm following SAH and reduces infarct size in ischemic models of stroke. PMID- 15831443 TI - Role of transient receptor potential vanilloid 1 receptors in adjuvant-induced chronic arthritis: in vivo study using gene-deficient mice. AB - The transient receptor potential vanilloid 1 (TRPV1) receptor is a nonselective cation channel localized on a subset of primary sensory neurons and can be activated by a wide range of stimuli. The present study investigated the role of this receptor in chronic arthritis evoked by complete Freund's adjuvant (CFA) using TRPV1 receptor gene-deleted (TRPV1-/-) mice and wild-type counterparts (TRPV1+/+). In TRPV1+/+ mice, CFA injected intraplantarly into the left hindpaw and the root of the tail induced swelling of the injected and contralateral paws up to 130 and 28%, respectively, measured by plethysmometry throughout 18 days. Mechanonociceptive threshold measured with dynamic plantar aesthesiometry was decreased by 50 and 18% on the injected and contralateral paws, respectively. Histological examination and scoring of the tibiotarsal joints revealed marked arthritic changes in wild-type mice. In TRPV1-/- animals edema, histological score and mechanical allodynia were significantly smaller. Daily treatment with the lipoxygenase inhibitor nordihydroguaretic acid (NDGA), the cyclooxygenase inhibitor indomethacin, the bradykinin B2 receptor antagonist HOE-140 [D-arginyl L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thyenyl)-L-alanyl-L-seryl D-1,2,2,4-tetrahydro-3-isoquinolinecarbonyl-L-(2a,3b,7ab)-octahydro-1H-indole-2 carbonyl-L-arginine], or the B1 receptor antagonist desArgHOE-140 [D-arginyl-L arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thyenyl)-L-alanyl-L-seryl-D 1,2,2,4-tetrahydro-3-isoquinolinecarbonyl-L-(2a,3b,7ab)-octahydro-1H-indole-2 carbonyl] was performed to reveal what mediators might activate TRPV1. NDGA significantly inhibited edema, hyperalgesia, and arthritis score in TRPV1+/+, but not in TRPV1-/- mice. The effect of indomethacin was markedly smaller in knockouts. In TRPV1+/+ animals, HOE-140, but not desArgHOE-140, inhibited arthritis, whereas in TRPV1-/- mice, HOE-140 produced limited effect. Thus, whereas bradykinin and lipoxygenase products seem to act exclusively via TRPV1 activation, prostanoids do not, or at least only partially, to enhance murine experimental arthritis and related hyperalgesia. PMID- 15831444 TI - Delta9-tetrahydrocannbinol accounts for the antinociceptive, hypothermic, and cataleptic effects of marijuana in mice. AB - Although it is widely accepted that delta9-tetrahydrocannabinol (delta9-THC) is the primary psychoactive constituent of marijuana, questions persist as to whether other components contribute to marijuana's pharmacological activity. The present experiments assessed the cannabinoid activity of marijuana smoke exposure in mice and tested the hypothesis that delta9-THC mediates these effects through a CB1 receptor mechanism of action. First, the effects of delta9-THC on analgesia, hypothermia, and catalepsy were compared with those of a marijuana extract with equated delta9-THC content after either i.v. administration or inhalation exposure. Second, mice were exposed to smoke of an ethanol-extracted placebo plant material or low-grade marijuana (with minimal delta9-THC but similar levels of other cannabinoids) that were impregnated with varying quantities of delta9-THC. To assess doses, delta9-THC levels in the blood and brains of drug-exposed mice were determined following both i.v. and inhalation routes of administration. Both marijuana and delta9-THC produced comparable levels of antinociception, hypothermia, and catalepsy regardless of the route of administration, and these effects were blocked by pretreatment with the CB1 antagonist SR141716 [N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl) 4-methyl-1H-pyrazole-3-carboxamide HCl]. Importantly, the blood and brain levels of delta9-THC were similar in mice exhibiting similar pharmacological effects, regardless of the presence of non-delta9-THC marijuana constituents. The present experiments provide evidence that the acute cannabinoid effects of marijuana smoke exposure on analgesia, hypothermia, and catalepsy in mice result from delta9-THC content acting at CB1 receptors and that the non-delta9-THC constituents of marijuana (at concentrations relevant to those typically consumed) influence these effects only minimally, if at all. PMID- 15831445 TI - Comments on "Memantine blocks alpha7* nicotinic acetylcholine receptors more potently than N-methyl-D-aspartate receptors in rat hippocampal neurons". PMID- 15831447 TI - Whole-genome analysis reveals a strong positional bias of conserved dMyc dependent E-boxes. AB - Myc is a transcription factor with diverse biological effects ranging from the control of cellular proliferation and growth to the induction of apoptosis. Here we present a comprehensive analysis of the transcriptional targets of the sole Myc ortholog in Drosophila melanogaster, dMyc. We show that the genes that are down-regulated in response to dmyc inhibition are largely identical to those that are up-regulated after dMyc overexpression and that many of them play a role in growth control. The promoter regions of these targets are characterized by the presence of the E-box sequence CACGTG, a known dMyc binding site. Surprisingly, a large subgroup of (functionally related) dMyc targets contains a single E-box located within the first 100 nucleotides after the transcription start site. The relevance of this E-box and its position was confirmed by a mutational analysis of a selected dMyc target and by the observation of its evolutionary conservation in a different Drosophila species, Drosophila pseudoobscura. These observations raise the possibility that a subset of Myc targets share a distinct regulatory mechanism. PMID- 15831448 TI - cis-regulatory logic of short-range transcriptional repression in Drosophila melanogaster. AB - Bioinformatics analysis of transcriptional control is guided by knowledge of the characteristics of cis-regulatory regions or enhancers. Features such as clustering of binding sites and co-occurrence of binding sites have aided enhancer identification, but quantitative predictions of enhancer function are not yet generally feasible. To facilitate the analysis of regulatory sequences in Drosophila melanogaster, we identified quantitative parameters that affect the activity of short-range transcriptional repressors, proteins that play key roles in development. In addition to the previously noted distance dependence, repression is strongly influenced by the stoichiometry, affinity, spacing, and arrangement of activator binding sites. Repression is insensitive to the type of activation domain, suggesting that short-range repression may primarily affect activators at the level of DNA binding. The activity of several short-range, but not long-range, repressors is circumscribed by the same quantitative parameters. This cis-regulatory "grammar" may aid the identification of enhancers regulated by short-range repressors and facilitate bioinformatic prediction of the functional output of transcriptional regulatory sequences. PMID- 15831449 TI - SPBP is a phosphoserine-specific repressor of estrogen receptor alpha. AB - Multiple signaling pathways stimulate the activity of estrogen receptor alpha (ERalpha) by direct phosphorylation within its N-terminal activation function 1 (AF1). How phosphorylation affects AF1 activity remains poorly understood. We performed a phage display screen for human proteins that are exclusively recruited to the phosphorylated form of AF1 and found the stromelysin-1 platelet derived growth factor-responsive element-binding protein (SPBP). In a purified system, SPBP bound only the in vitro-phosphorylated form of the ERalpha AF1 or the phosphoserine mimic S118E, and the interaction domain could be mapped to a 42 amino-acid fragment of SPBP. In cells, SPBP preferentially interacted with liganded and phosphorylated ERalpha. Functionally, SPBP behaved as a repressor of activated ERalpha, which extends its previously demonstrated roles as a DNA binding transactivation factor and coactivator of other transcription factors. By targeting the phosphorylated form of AF1, SPBP may contribute to attenuating and fine-tuning ERalpha activity. A functional consequence is that SPBP inhibits the proliferation of ERalpha-dependent but not ERalpha-independent breast cancer cell lines, mirroring a reported negative correlation with the ERalpha status of breast tumors. PMID- 15831450 TI - Recql5 and Blm RecQ DNA helicases have nonredundant roles in suppressing crossovers. AB - In eukaryotes, crossovers in mitotic cells can have deleterious consequences and therefore must be suppressed. Mutations in BLM give rise to Bloom syndrome, a disease that is characterized by an elevated rate of crossovers and increased cancer susceptibility. However, simple eukaryotes such as Saccharomyces cerevisiae have multiple pathways for suppressing crossovers, suggesting that mammals also have multiple pathways for controlling crossovers in their mitotic cells. We show here that in mouse embryonic stem (ES) cells, mutations in either the Bloom syndrome homologue (Blm) or the Recql5 genes result in a significant increase in the frequency of sister chromatid exchange (SCE), whereas deleting both Blm and Recql5 lead to an even higher frequency of SCE. These data indicate that Blm and Recql5 have nonredundant roles in suppressing crossovers in mouse ES cells. Furthermore, we show that mouse embryonic fibroblasts derived from Recql5 knockout mice also exhibit a significantly increased frequency of SCE compared with the corresponding wild-type control. Thus, this study identifies a previously unknown Recql5-dependent, Blm-independent pathway for suppressing crossovers during mitosis in mice. PMID- 15831451 TI - Adult stage gamma-globin silencing is mediated by a promoter direct repeat element. AB - The human beta-like globin genes (5'-epsilon-Ggamma-Agamma-delta-beta-3') are temporally expressed in sequential order from the 5' to 3' end of the locus, but the nonadult epsilon- and gamma-globin genes are autonomously silenced in adult erythroid cells. Two cis elements have been proposed to regulate definitive erythroid gamma-globin repression: the DR (direct repeat) and CCTTG elements. Since these two elements partially overlap, and since a well-characterized HPFH point mutation maps to an overlapping nucleotide, it is not clear if both or only one of the two participate in gamma-globin silencing. To evaluate the contribution of these hypothetical silencers to gamma-globin regulation, we generated point mutations that individually disrupted either the single DR or all four CCTTG elements. These two were separately incorporated into human beta globin yeast artificial chromosomes, which were then used to generate gamma globin mutant transgenic mice. While DR element mutation led to a dramatic increase in Agamma-globin expression only during definitive erythropoiesis, the CCTTG mutation did not affect adult stage transcription. These results demonstrate that the DR sequence element autonomously mediates definitive stage specific gamma-globin gene silencing. PMID- 15831452 TI - Sequence-specific DNA binding by the alphaNAC coactivator is required for potentiation of c-Jun-dependent transcription of the osteocalcin gene. AB - Since the c-Jun coactivator alphaNAC was initially identified in a differential screen for genes expressed in differentiated osteoblasts, we examined whether the osteocalcin gene, a specific marker of terminal osteoblastic differentiation, could be a natural target for the coactivating function of alphaNAC. We had also previously shown that alphaNAC can specifically bind DNA in vitro, but it remained unclear whether the DNA-binding function of alphaNAC is expressed in vivo or if it is required for coactivation. We have identified an alphaNAC binding site within the murine osteocalcin gene proximal promoter region and demonstrated that recombinant alphaNAC or alphaNAC from ROS17/2.8 nuclear extracts can specifically bind this element. Using transient transfection assays, we have shown that alphaNAC specifically potentiated the c-Jun-dependent transcription of the osteocalcin promoter and that this activity specifically required the DNA-binding domain of alphaNAC. Chromatin immunoprecipitation confirmed that alphaNAC occupies its binding site on the osteocalcin promoter in living osteoblastic cells expressing osteocalcin. Inhibition of the expression of endogenous alphaNAC in osteoblastic cells by use of RNA interference provoked a decrease in osteocalcin gene transcription. Our results show that the osteocalcin gene is a target for the alphaNAC coactivating function, and we propose that alphaNAC is specifically targeted to the osteocalcin promoter through its DNA binding activity as a means to achieve increased specificity in gene transcription. PMID- 15831453 TI - Interdependent recruitment of SAGA and Srb mediator by transcriptional activator Gcn4p. AB - Transcriptional activation by Gcn4p is enhanced by the coactivators SWI/SNF, SAGA, and Srb mediator, which stimulate recruitment of TATA binding protein (TBP) and polymerase II to target promoters. We show that wild-type recruitment of SAGA by Gcn4p is dependent on mediator but independent of SWI/SNF function at three different promoters. Recruitment of mediator is also independent of SWI/SNF but is enhanced by SAGA at a subset of Gcn4p target genes. Recruitment of all three coactivators to ARG1 is independent of the TATA element and preinitiation complex formation, whereas efficient recruitment of the general transcription factors requires the TATA box. We propose an activation pathway involving interdependent recruitment of SAGA and Srb mediator to the upstream activation sequence, enabling SWI/SNF recruitment and the binding of TBP and other general factors to the promoter. We also found that high-level recruitment of Tra1p and other SAGA subunits is independent of the Ada2p/Ada3p/Gcn5p histone acetyltransferase module but requires Spt3p in addition to subunits required for SAGA integrity. Thus, while Tra1p can bind directly to Gcn4p in vitro, it requires other SAGA subunits for efficient recruitment in vivo. PMID- 15831454 TI - The mechanism of endogenous receptor activation functionally distinguishes prototype canonical and noncanonical Wnts. AB - Wnt glycoproteins are developmentally essential signaling molecules, and lesions afflicting Wnt pathways play important roles in human diseases. Some Wnts signal to the canonical pathway by stabilizing beta-catenin, while others lack this activity. Frizzled serpentine receptors mediate distinct signaling pathways by both classes of Wnts. Here, we tandemly linked noncanonical Wnt5a with the C terminal half of Dickkopf-2 (Dkk2C), a distinct ligand of the Wnt coreceptor LRP5/6. Whereas Wnt5a, Dkk2C, or both together were incapable of stimulating endogenous canonical signaling, the Wnt5a/Dkk2C chimera efficiently activated this pathway in a manner inhibitable by specific antagonists of either frizzled or LRP receptors. Thus, activation of the canonical pathway requires ligand coupling of an endogenous frizzled/LRP coreceptor complex, rather than Wnt triggering each receptor independently. Moreover, fusion of Wnt5a with Dkk2C unmasked its ability to signal to Dishevelled through multiple frizzleds, indicating that the lack of functional interaction with LRP distinguishes noncanonical Wnt5a from canonical Wnts in mammalian cells. These findings provide a novel mechanism by which the same receptor can be switched between distinct signaling pathways depending on the differential recruitment of a coreceptor by members of the same ligand family. PMID- 15831455 TI - FAT10, a ubiquitin-independent signal for proteasomal degradation. AB - FAT10 is a small ubiquitin-like modifier that is encoded in the major histocompatibility complex and is synergistically inducible by tumor necrosis factor alpha and gamma interferon. It is composed of two ubiquitin-like domains and possesses a free C-terminal diglycine motif that is required for the formation of FAT10 conjugates. Here we show that unconjugated FAT10 and a FAT10 conjugate were rapidly degraded by the proteasome at a similar rate. Fusion of FAT10 to the N terminus of very long-lived proteins enhanced their degradation rate as potently as fusion with ubiquitin did. FAT10-green fluorescent protein fusion proteins were not cleaved but entirely degraded, suggesting that FAT10 specific deconjugating enzymes were not present in the analyzed cell lines. Interestingly, the prevention of ubiquitylation of FAT10 by mutation of all lysines or by expression in ubiquitylation-deficient cells did not affect FAT10 degradation. Thus, conjugation with FAT10 is an alternative and ubiquitin independent targeting mechanism for degradation by the proteasome, which, in contrast to polyubiquitylation, is cytokine inducible and irreversible. PMID- 15831456 TI - Orphan nuclear receptor LRH-1 is required to maintain Oct4 expression at the epiblast stage of embryonic development. AB - Oct4 plays an essential role in maintaining the inner cell mass and pluripotence of embryonic stem (ES) cells. The expression of Oct4 is regulated by the proximal enhancer and promoter in the epiblast and by the distal enhancer and promoter at all other stages in the pluripotent cell lineage. Here we report that the orphan nuclear receptor LRH-1, which is expressed in undifferentiated ES cells, can bind to SF-1 response elements in the proximal promoter and proximal enhancer of the Oct4 gene and activate Oct4 reporter gene expression. LRH-1 is colocalized with Oct4 in the inner cell mass and the epiblast of embryos at early developmental stages. Disruption of the LRH-1 gene results in loss of Oct4 expression at the epiblast stage and early embryonic death. Using LRH-1(-/-) ES cells, we also show that LRH-1 is required to maintain Oct4 expression at early differentiation time points. In vitro and in vivo results show that LRH-1 plays an essential role in the maintenance of Oct4 expression in ES cells at the epiblast stage of embryonic development, thereby maintaining pluripotence at this crucial developmental stage prior to segregation of the primordial germ cell lineage at gastrulation. PMID- 15831457 TI - SUMO-1 modification of PIASy, an E3 ligase, is necessary for PIASy-dependent activation of Tcf-4. AB - We have previously shown that modification of Tcf-4, a transcription factor in the Wnt pathway, with SUMO by PIASy, a SUMO E3 ligase, enhances its transcriptional activity. Since PIASy itself was also modified with SUMO-1, we studied the role of sumoylation of PIASy in the regulation of Tcf-4. Lys(35) was found to be a sumoylation site of PIASy. PIASy(K35R), in which Lys(35) was mutated to Arg, did not enhance sumoylation of Tcf-4, although this PIASy mutant did not lose the ligase activity of sumoylation for other proteins. Wild-type PIASy and PIASy(K35R) showed a distinct distribution in the nucleus, although both were colocalized with Tcf-4. Promyelocytic leukemia protein, which is involved in transcriptional regulation, was associated with PIASy(K35R) more frequently than wild-type PIASy in the nucleus. PIASy(K35R) could not stimulate the transcriptional activity of Tcf-4 under the conditions in which wild-type PIASy enhanced it. Conjugation of SUMO-1 to the amino terminus of PIASy(K35R) neither enhanced sumoylation of Tcf-4 nor stimulated the transcriptional activity of Tcf-4. These results suggest that sumoylation of Lys(35) in PIASy determines the nuclear localization of PIASy and that it is necessary for PIASy-dependent sumoylation and transcriptional activation of Tcf-4. PMID- 15831458 TI - The pleckstrin homology domain-containing protein CKIP-1 is involved in regulation of cell morphology and the actin cytoskeleton and interaction with actin capping protein. AB - CKIP-1 is a pleckstrin homology domain-containing protein that interacts with protein kinase CK2. To elucidate the functions of CKIP-1, we generated human osteosarcoma cell lines with tetracycline-regulated expression of Flag-CKIP-1. Flag-CKIP-1 expression resulted in distinct changes in cellular morphology. Therefore, we examined the actin profile by immunofluorescence, quantitative measurement of phalloidin binding, and immunoblot analysis. These studies demonstrate that Flag-CKIP-1 expression resulted in increases in F-actin staining and protein levels of beta-actin. To elucidate the mechanisms behind the observed phenotype, we utilized tandem affinity purification to isolate CKIP-1 interacting proteins. Mass spectrometry analysis led to the identification of the actin capping protein subunits, CPalpha and CPbeta, as novel CKIP-1 interaction partners. Interactions were confirmed by coimmunoprecipitation and by colocalization. Furthermore, we demonstrate that Ser9 of CPalpha is phosphorylated by protein kinase CK2 in vitro, that CPalpha is phosphorylated in vivo, and that treatment with a CK2-specific inhibitor results in a decrease in CPalpha phosphorylation. Finally, we demonstrate that CKIP-1 and CK2 inhibit the activity of actin capping protein at the barbed ends of actin filaments. Overall, our results are consistent with CKIP-1 playing a role in the regulation of the actin cytoskeleton through its interactions with actin capping protein. PMID- 15831459 TI - Inactivation of CtIP leads to early embryonic lethality mediated by G1 restraint and to tumorigenesis by haploid insufficiency. AB - CtIP interacts with a group of tumor suppressor proteins including RB (retinoblastoma protein), BRCA1, Ikaros, and CtBP, which regulate cell cycle progression through transcriptional repression as well as chromatin remodeling. However, how CtIP exerts its biological function in cell cycle progression remains elusive. To address this issue, we generated an inactivated Ctip allele in mice by inserting a neo gene into exon 5. The corresponding Ctip(-/-) embryos died at embryonic day 4.0 (E4.0), and the blastocysts failed to enter S phase but accumulated in G(1), leading to a slightly elevated cell death. Mouse NIH 3T3 cells depleted of Ctip were arrested at G(1) with the concomitant increase in hypophosphorylated Rb and Cdk inhibitors, p21. However, depletion of Ctip failed to arrest Rb(-/-) mouse embryonic fibroblasts (MEF) or human osteosarcoma Saos-2 cells at G(1), suggesting that this arrest is RB dependent. Importantly, the life span of Ctip(+/-) heterozygotes was shortened by the development of multiple types of tumors, predominantly, large lymphomas. The wild-type Ctip allele and protein remained detectable in these tumors, suggesting that haploid insufficiency of Ctip leads to tumorigenesis. Taken together, this finding uncovers a novel G(1)/S regulation in that CtIP counteracts Rb-mediated G(1) restraint. Deregulation of this function leads to a defect in early embryogenesis and contributes, in part, to tumor formation. PMID- 15831460 TI - Identification in vivo of different rate-limiting steps associated with transcriptional activators in the presence and absence of a GAGA element. AB - We analyzed the impact of a GAGA element on a transgenic promoter in Drosophila melanogaster that was activated by proteins composed of the Tet(on) DNA binding domain and either the heat shock factor (HSF) activation domain or a potent subdomain of VP16. Permanganate footprinting was used to monitor polymerase II (Pol II) on the transgenic promoters in vivo. Activation by Tet(on)-HSF but not by Tet(on)-VP16(A2) required the GAGA element; this correlated with the ability of the GAGA element to establish a paused Pol II. Although the GAGA element was not required for activation by Tet(on)-VP16(A2), the GAGA element greatly accelerated the rate of activation. The permanganate data also provided evidence that Pol II encountered different rate-limiting steps, following initiation in the presence of Tet(on)-HSF and Tet(on)-VP16(A2). The rate-limiting step in the presence of Tet(on)-HSF was release of Pol II paused about 20 to 40 nucleotides downstream from the start site. The rate-limiting step in the presence of Tet(on) VP16(A2) occurred much closer to the transcription start site. Several biochemical studies have provided evidence for a structural transition shortly after Pol II initiates transcription. The behavior of Pol II in the presence of Tet(on)-VP16(A2) provides the first evidence that this transition occurs in vivo. PMID- 15831461 TI - Inhibition of human Chk1 causes increased initiation of DNA replication, phosphorylation of ATR targets, and DNA breakage. AB - Human checkpoint kinase 1 (Chk1) is an essential kinase required to preserve genome stability. Here, we show that Chk1 inhibition by two distinct drugs, UCN 01 and CEP-3891, or by Chk1 small interfering RNA (siRNA) leads to phosphorylation of ATR targets. Chk1-inhibition triggered rapid, pan-nuclear phosphorylation of histone H2AX, p53, Smc1, replication protein A, and Chk1 itself in human S-phase cells. These phosphorylations were inhibited by ATR siRNA and caffeine, but they occurred independently of ATM. Chk1 inhibition also caused an increased initiation of DNA replication, which was accompanied by increased amounts of nonextractable RPA protein, formation of single-stranded DNA, and induction of DNA strand breaks. Moreover, these responses were prevented by siRNA mediated downregulation of Cdk2 or the replication initiation protein Cdc45, or by addition of the CDK inhibitor roscovitine. We propose that Chk1 is required during normal S phase to avoid aberrantly increased initiation of DNA replication, thereby protecting against DNA breakage. These results may help explain why Chk1 is an essential kinase and should be taken into account when drugs to inhibit this kinase are considered for use in cancer treatment. PMID- 15831462 TI - Id2 mediates tumor initiation, proliferation, and angiogenesis in Rb mutant mice. AB - The inhibitor of differentiation Id2 is a target of the retinoblastoma (Rb) protein during mouse embryogenesis. In Rb(+/-) mice, LOH at the wild-type Rb allele initiates pituitary adenocarcinoma, a tumor derived from embryonic melanotropes. Here we identify a critical role for Id2 in initiation, growth, and angiogenesis of pituitary tumors from Rb(+/-) mice. We show that proliferation and differentiation are intimately coupled in Rb(+/-) pituitary cells before tumor initiation. In Id2-null pituitaries, premature activation of basic helix loop-helix-mediated transcription and expression of the cdk inhibitor p27(Kip1) impairs the proliferation of melanotropes and tumor initiation. Without Id2, Rb(+/-) mice have fewer early tumor lesions and a markedly decreased proliferation rate of the tumor foci. Expression of Id2 by pituitary tumor cells promotes growth and angiogenesis by functioning as a master regulator of vascular endothelial growth factor (VEGF). In human neuroblastoma, the N-Myc-driven expression of Id2 is sufficient and necessary for expression of VEGF. These results establish that aberrant Id2 activity directs initiation and progression of embryonal cancer. PMID- 15831463 TI - Myocyte enhancer factor 2 acetylation by p300 enhances its DNA binding activity, transcriptional activity, and myogenic differentiation. AB - Myocyte enhancer factor 2 (MEF2) family proteins are key transcription factors controlling gene expression in myocytes, lymphocytes, and neurons. MEF2 proteins are known to be regulated by phosphorylation. We now provide evidence showing that MEF2C is acetylated by p300 both in vitro and in vivo. In C2C12 myogenic cells, MEF2 is preferentially acetylated in differentiating myocytes but not in undifferentiated myoblasts. Several major acetylation sites are mapped to the transactivation domain of MEF2C, some of which are fully conserved in other MEF2 members from several different species. Mutation of these lysines affects MEF2 DNA binding and transcriptional activity, as well as its synergistic effect with myogenin in myogenic conversion assays. When introduced into C2C12 myoblasts, the nonacetylatable MEF2C inhibits myogenic differentiation. Thus, in addition to phosphorylation, MEF2 activity is also critically regulated by acetylation during myogenesis. PMID- 15831464 TI - Human RNA polymerase II elongation in slow motion: role of the TFIIF RAP74 alpha1 helix in nucleoside triphosphate-driven translocation. AB - The role of the RAP74 alpha1 helix of transcription factor IIF (TFIIF) in stimulating elongation by human RNA polymerase II (RNAP II) was examined using millisecond-phase transient-state kinetics. RAP74 deletion mutants RAP74(1-227), which includes an intact alpha1 helix, and RAP74(1-158), in which the alpha1 helix is deleted, were compared. Analysis of TFIIF RAP74-RAP30 complexes carrying the RAP74(1-158) deletion reveals the role of the alpha1 helix because this mutant has indistinguishable activity compared to TFIIF 74(W164A), which carries a critical point mutation in alpha1. We report adequate two-bond kinetic simulations for the reaction in the presence of TFIIF 74(1-227) + TFIIS and TFIIF 74(1-158) + TFIIS. TFIIF 74(1-158) is defective because it fails to promote forward translocation. Deletion of the RAP74 alpha1 helix results in increased occupancy of the backtracking, cleavage, and restart pathways at a stall position, indicating reverse translocation of the elongation complex. During elongation, TFIIF 74(1-158) fails to support detectable nucleoside triphosphate (NTP)-driven translocation from a stall position and is notably defective in supporting bond completion (NTP-driven translocation coupled to pyrophosphate release) during the processive transition between bonds. PMID- 15831465 TI - Mismatch repair proteins are activators of toxic responses to chromium-DNA damage. AB - Chromium(VI) is a toxic and carcinogenic metal that causes the formation of DNA phosphate-based adducts. Cr-DNA adducts are genotoxic in human cells, although they do not block replication in vitro. Here, we report that induction of cytotoxicity in Cr(VI)-treated human colon cells and mouse embryonic fibroblasts requires the presence of all major mismatch repair (MMR) proteins. Cr-DNA adducts lost their ability to block replication of Cr-modified plasmids in human colon cells lacking MLH1 protein. The presence of functional mismatch repair caused induction of p53-independent apoptosis associated with activation of caspases 2 and 7. Processing of Cr-DNA damage by mismatch repair resulted in the extensive formation of gamma-H2AX foci in G(2) phase, indicating generation of double stranded breaks as secondary toxic lesions. Induction of gamma-H2AX foci was observed at 6 to 12 h postexposure, which was followed by activation of apoptosis in the absence of significant G(2) arrest. Our results demonstrate that mismatch repair system triggers toxic responses to Cr-DNA backbone modifications through stress mechanisms that are significantly different from those for other forms of DNA damage. Selection for Cr(VI) resistant, MMR-deficient cells may explain the very high frequency of lung cancers with microsatellite instability among chromate workers. PMID- 15831466 TI - Induction of ectopic olfactory structures and bone morphogenetic protein inhibition by Rossy, a group XII secreted phospholipase A2. AB - The secreted phospholipases A(2) (sPLA(2)s) comprise a family of small secreted proteins with the ability to catalyze the generation of bioactive lipids through glycophospholipid hydrolysis. Recently, a large number of receptor proteins and extracellular binding partners for the sPLA(2)s have been identified, suggesting that these secreted factors might exert a subset of their broad spectrum of biological activities independently of their enzymatic activity. Here, we describe an activity for the sPLA(2) group XII (sPLA(2)-gXII) gene during Xenopus laevis early development. In the ectoderm, sPLA(2)-gXII acts as a neural inducer by blocking bone morphogenetic protein (BMP) signaling. Gain of function in embryos leads to ectopic neurogenesis and to the specification of ectopic olfactory sensory structures, including olfactory bulb and sensory epithelia. This activity is conserved in the Drosophila melanogaster, Xenopus, and mammalian orthologs and appears to be independent of the lipid hydrolytic activity. Because of its effect on olfactory neurogenesis, we have renamed this gene Rossy, in homage to the Spanish actress Rossy de Palma. We present evidence that Rossy/sPLA(2)-gXII can inhibit the transcriptional activation of BMP direct target gene reporters in Xenopus and mouse P19 embryonic carcinoma cells through the loss of DNA-binding activity of activated Smad1/4 complexes. Collectively, these data represent the first evidence for signaling cross talk between a secreted phospholipase A(2) and the BMP/transforming growth factor beta pathways and identify Rossy/sPLA(2)-gXII as the only factor thus far described which is sufficient to induce anterior sensory neural structures during vertebrate development. PMID- 15831467 TI - Caspase-3-dependent beta-cell apoptosis in the initiation of autoimmune diabetes mellitus. AB - beta-Cell apoptosis is a key event contributing to the pathogenesis of type 1 diabetes mellitus. In addition to apoptosis being the main mechanism by which beta cells are destroyed, beta-cell apoptosis has been implicated in the initiation of type 1 diabetes mellitus through antigen cross-presentation mechanisms that lead to beta-cell-specific T-cell activation. Caspase-3 is the major effector caspase involved in apoptotic pathways. Despite evidence supporting the importance of beta-cell apoptosis in the pathogenesis of type 1 diabetes, the specific role of caspase-3 in this process is unknown. Here, we show that Caspase-3 knockout (Casp3(-/-) mice were protected from developing diabetes in a multiple-low-dose streptozotocin autoimmune diabetes model. Lymphocyte infiltration of the pancreatic islets was completely absent in Casp3( /-) mice. To determine the role of caspase-3-dependent apoptosis in disease initiation, a defined antigen-T-cell receptor transgenic system, RIP-GP/P14 double-transgenic mice with Casp3 null mutation, was examined. beta-cell antigen specific T-cell activation and proliferation were observed only in the pancreatic draining lymph node of RIP-GP/P14/Casp3(+/-) mice, but not in mice lacking caspase-3. Together, our findings demonstrate that caspase-3-mediated beta-cell apoptosis is a requisite step for T-cell priming, a key initiating event in type 1 diabetes. PMID- 15831468 TI - The phosphatidylinositol 3-phosphate phosphatase myotubularin- related protein 6 (MTMR6) is a negative regulator of the Ca2+-activated K+ channel KCa3.1. AB - Myotubularins (MTMs) belong to a large subfamily of phosphatases that dephosphorylate the 3' position of phosphatidylinositol 3-phosphate [PI(3)P] and PI(3,5)P(2). MTM1 is mutated in X-linked myotubular myopathy, and MTMR2 and MTMR13 are mutated in Charcot-Marie-Tooth syndrome. However, little is known about the general mechanism(s) whereby MTMs are regulated or the specific biological processes regulated by the different MTMs. We identified a Ca(2+) activated K channel, K(Ca)3.1 (also known as KCa4, IKCa1, hIK1, or SK4), that specifically interacts with the MTMR6 subfamily of MTMs via coiled coil (CC) domains on both proteins. Overexpression of MTMR6 inhibited K(Ca)3.1 channel activity, and this inhibition required MTMR6's CC and phosphatase domains. This inhibition is specific; MTM1, a closely related MTM, did not inhibit K(Ca)3.1. However, a chimeric MTM1 in which the MTM1 CC domain was swapped for the MTMR6 CC domain inhibited K(Ca)3.1, indicating that MTM CC domains are sufficient to confer target specificity. K(Ca)3.1 was also inhibited by the PI(3) kinase inhibitors LY294002 and wortmannin, and this inhibition was rescued by the addition of PI(3)P, but not other phosphoinositides, to the patch pipette solution. PI(3)P also rescued the inhibition of K(Ca)3.1 by MTMR6 overexpression. These data, when taken together, indicate that K(Ca)3.1 is regulated by PI(3)P and that MTMR6 inhibits K(Ca)3.1 by dephosphorylating the 3' position of PI(3)P, possibly leading to decreased PI(3)P in lipid microdomains adjacent to K(Ca)3.1. K(Ca)3.1 plays important roles in controlling proliferation by T cells, vascular smooth muscle cells, and some cancer cell lines. Thus, our findings not only provide unique insights into the regulation of K(Ca)3.1 channel activity but also raise the possibility that MTMs play important roles in the negative regulation of T cells and in conditions associated with pathological cell proliferation, such as cancer and atherosclerosis. PMID- 15831470 TI - Integrin-linked kinase mediates bone morphogenetic protein 7-dependent renal epithelial cell morphogenesis. AB - Bone morphogenetic protein 7 (BMP7) stimulates renal branching morphogenesis via p38 mitogen-activated protein kinase (p38(MAPK)) and activating transcription factor 2 (ATF-2) (M. C. Hu, D. Wasserman, S. Hartwig, and N. D. Rosenblum, J. Biol. Chem. 279:12051-12059, 2004). Here, we demonstrate a novel role for integrin-linked kinase (ILK) in mediating renal epithelial cell morphogenesis in embryonic kidney explants and identify p38(MAPK) as a target of ILK signaling in a cell culture model of renal epithelial morphogenesis. The spatial and temporal expression of ILK in embryonic mouse kidney cells suggested a role in branching morphogenesis. Adenovirus-mediated expression of ILK stimulated and expression of a dominant negative ILK mutant inhibited ureteric bud branching in embryonic mouse kidney explants. BMP7 increased ILK kinase activity in inner medullary collecting duct 3 (IMCD-3) cells, and adenovirus-mediated expression of ILK increased IMCD-3 cell morphogenesis in a three-dimensional culture model. In contrast, treatment with a small molecule ILK inhibitor or expression of a dominant negative-acting ILK (ILK(E359K)) inhibited epithelial cell morphogenesis. Further, expression of ILK(E359K) abrogated BMP7-dependent stimulation. To investigate the role of ILK in BMP7 signaling, we showed that ILK overexpression increased basal and BMP7-induced levels of phospho-p38(MAPK) and phospho-ATF-2. Consistent with its inhibitory effects on IMCD-3 cell morphogenesis, expression of ILK(E359K) blocked BMP7-dependent increases in phospho-p38(MAPK) and phospho-ATF-2. Inhibition of p38(MAPK) activity with the specific inhibitor, SB203580, failed to inhibit BMP7-dependent stimulation of ILK activity, suggesting that ILK functions upstream of p38(MAPK) during BMP7 signaling. We conclude that ILK functions in a BMP7/p38(MAPK)/ATF-2 signaling pathway and stimulates epithelial cell morphogenesis. PMID- 15831469 TI - Targeted disruption of Tgif, the mouse ortholog of a human holoprosencephaly gene, does not result in holoprosencephaly in mice. AB - 5'-TG-3'-interacting factor or transforming growth factor beta (TGF-beta)-induced factor (TGIF) belongs to a family of evolutionarily conserved proteins that are characterized by an atypical three-amino-acid loop extension homeodomain. In vitro studies have implicated TGIF as a transcriptional repressor and corepressor in retinoid and TGF-beta signaling pathways that regulate several important biological processes. Heterozygous nonsense and missense mutations of the human TGIF gene have been associated with holoprosencephaly, the most common congenital malformation of the forebrain. In mice, Tgif mRNA is expressed ubiquitously in the ventricular neuroepithelium at embryonic day 10.5 (E10.5) but displays a medial to lateral gradient in the developing cerebral cortex at E12.5. The expression quickly declines by E14.5. The spatiotemporal expression profile of Tgif is consistent with its involvement in midline forebrain development. To better understand the function of Tgif in forebrain patterning and proliferation in vivo, we generated mice lacking Tgif by targeted deletion of exons 2 and 3, which encode 98% of the amino acids. Tgif(-)(/)(-) mice had no detectable Tgif protein by Western blotting. Surprisingly, however, these mice were viable and fertile. In addition, there were no discernible derangements in any of the major organ systems, including the forebrain. Overall our results point to a possible functional redundancy of Tgif, potentially provided by the closely related Tgif2. PMID- 15831471 TI - The distal sequence element of the selenocysteine tRNA gene is a tissue-dependent enhancer essential for mouse embryogenesis. AB - Appropriate expression of the selenocysteine tRNA (tRNA(Sec)) gene is necessary for the production of an entire family of selenoprotein enzymes. This study investigates the consequence of disrupting an upstream enhancer region of the mouse tRNA(Sec) gene (Trsp) known as the distal sequence element (DSE) by use of a conditional repair gene targeting strategy, in which a 3.2-kb insertion was introduced into the promoter of the gene. In the absence of DSE activity, homozygous mice failed to develop in utero beyond embryonic day 7.5 and had severely decreased levels of selenoprotein transcript. Cre-mediated removal of the selection cassette recovered DSE regulation of Trsp, restoring wild-type levels of tRNA(Sec) expression and allowing the generation of viable rescued mice. Further analysis of targeted heterozygous adult mice revealed that the enhancer activity of the DSE is tissue dependent since, in contrast to liver, heart does not require the DSE for normal expression of Trsp. Similarly, in mouse cell lines we showed that the DSE functions as a cell-line-specific inducible element of tRNA(Sec). Together, our data demonstrate that the DSE is a tissue dependent regulatory element of tRNA(Sec) expression and that its activity is vital for sufficient tRNA(Sec) production during mouse embryogenesis. PMID- 15831472 TI - Role of MLK3 in the regulation of mitogen-activated protein kinase signaling cascades. AB - Mixed-lineage protein kinase 3 (MLK3) is a member of the mitogen-activated protein (MAP) kinase kinase kinase group that has been implicated in multiple signaling cascades, including the NF-kappaB pathway and the extracellular signal regulated kinase, c-Jun NH(2)-terminal kinase (JNK), and p38 MAP kinase pathways. Here, we examined the effect of targeted disruption of the murine Mlk3 gene. Mlk3(-/-) mice were found to be viable and healthy. Primary embryonic fibroblasts prepared from these mice exhibited no major signaling defects. However, we did find that MLK3 deficiency caused a selective reduction in tumor necrosis factor (TNF)-stimulated JNK activation. Together, these data demonstrate that MLK3 contributes to the TNF signaling pathway that activates JNK. PMID- 15831473 TI - The Mcp element from the bithorax complex contains an insulator that is capable of pairwise interactions and can facilitate enhancer-promoter communication. AB - Chromatin insulators, or boundary elements, appear to control eukaryotic gene expression by regulating interactions between enhancers and promoters. Boundaries have been identified in the 3' cis-regulatory region of Abd-B, which is subdivided into a series of separate iab domains. Boundary elements such as Mcp, Fab-7, and Fab-8 and adjacent silencers flank the iab domains and restrict the activity of the iab enhancers. We have identified an insulator in the 755-bp Mcp fragment that is linked to the previously characterized Polycomb response element (PRE) and silences the adjacent genes. This insulator blocks the enhancers of the yellow and white genes and protects them from PRE-mediated repression. The interaction between the Mcp elements, each containing the insulator and PRE, allows the eye enhancer to activate the white promoter over the repressed yellow domain. The same level of white activation was observed when the Mcp element combined with the insulator alone was interposed between the eye enhancer and the promoter, suggesting that the insulator is responsible for the interaction between the Mcp elements. PMID- 15831474 TI - Tyrosine phosphorylation regulates maturation of receptor tyrosine kinases. AB - Constitutive activation of receptor tyrosine kinases (RTKs) is a frequent event in human cancer cells. Activating mutations in Fms-like tyrosine kinase 3 (FLT 3), notably, internal tandem duplications in the juxtamembrane domain (FLT-3 ITD), have been causally linked to acute myeloid leukemia. As we describe here, FLT-3 ITD exists predominantly in an immature, underglycosylated 130-kDa form, whereas wild-type FLT-3 is expressed predominantly as a mature, complex glycosylated 150-kDa molecule. Endogenous FLT-3 ITD, but little wild-type FLT-3, is detectable in the endoplasmic reticulum (ER) compartment. Conversely, cell surface expression of FLT-3 ITD is less efficient than that of wild-type FLT-3. Inhibition of FLT-3 ITD kinase by small molecules, inactivating point mutations, or coexpression with the protein-tyrosine phosphatases (PTPs) SHP-1, PTP1B, and PTP-PEST but not RPTPalpha promotes complex glycosylation and surface localization. However, PTP coexpression has no effect on the maturation of a surface glycoprotein of vesicular stomatitis virus. The maturation of wild-type FLT-3 is impaired by general PTP inhibition or by suppression of endogenous PTP1B. Enhanced complex formation of FLT-3 ITD with the ER-resident chaperone calnexin indicates that its retention in the ER is related to inefficient folding. The regulation of RTK maturation by tyrosine phosphorylation was observed with other RTKs as well, defines a possible role for ER-resident PTPs, and may be related to the altered signaling quality of constitutively active, transforming RTK mutants. PMID- 15831475 TI - Caspase-dependent regulation and subcellular redistribution of the transcriptional modulator YY1 during apoptosis. AB - The transcriptional regulator Yin Yang 1 (YY1) controls many aspects of cell behavior and is essential for development. We analyzed the fate of YY1 during apoptosis and studied the functional consequences. We observed that this factor is rapidly translocated into the cell nucleus in response to various apoptotic stimuli, including activation of Fas, stimulation by tumor necrosis factor, and staurosporine and etoposide treatment. Furthermore, YY1 is cleaved by caspases in vitro and in vivo at two distinct sites, IATD(12)G and DDSD(119)G, resulting in the deletion of the first 119 amino acids early in the apoptotic process. This activity generates an N-terminally truncated YY1 fragment (YY1Delta119) that has lost its transactivation domain but retains its DNA binding domain. Indeed, YY1Delta119 is no longer able to stimulate gene transcription but interacts with DNA. YY1Delta119 but not the wild-type protein or the caspase-resistant mutant YY1D12A/D119A enhances Fas-induced apoptosis, suggesting that YY1 is involved in a positive feedback loop during apoptosis. Our findings provide evidence for a new mode of regulation of YY1 and define a novel aspect of the involvement of YY1 in the apoptotic process. PMID- 15831476 TI - Interaction of BAG1 and Hsp70 mediates neuroprotectivity and increases chaperone activity. AB - It was recently shown that Bcl-2-associated athanogene 1 (BAG1) is a potent neuroprotectant as well as a marker of neuronal differentiation. Since there appears to exist an equilibrium within the cell between BAG1 binding to heat shock protein 70 (Hsp70) and BAG1 binding to Raf-1 kinase, we hypothesized that changing BAG1 binding characteristics might significantly alter BAG1 function. To this end, we compared rat CSM14.1 cells and human SHSY-5Y cells stably overexpressing full-length BAG1 or a deletion mutant (BAGDeltaC) no longer capable of binding to Hsp70. Using a novel yellow fluorescent protein-based foldase biosensor, we demonstrated an upregulation of chaperone in situ activity in cells overexpressing full-length BAG1 but not in cells overexpressing BAGDeltaC compared to wild-type cells. Interestingly, in contrast to the nuclear and cytosolic localizations of full-length BAG1, BAGDeltaC was expressed exclusively in the cytosol. Furthermore, cells expressing BAGDeltaC were no longer protected against cell death. However, they still showed accelerated neuronal differentiation. Together, these results suggest that BAG1-induced activation of Hsp70 is important for neuroprotectivity, while BAG1-dependent modulation of neuronal differentiation in vitro is not. PMID- 15831477 TI - p21-activated kinase 1 regulates microtubule dynamics by phosphorylating tubulin cofactor B. AB - p21-activated kinase 1 (Pak1) induces cytoskeleton reorganization in part by regulating microtubule dynamics through an elusive mechanism. Using a yeast two hybrid screen, we identified tubulin cofactor B (TCoB) (a cofactor in the assembly of the alpha/beta-tubulin heterodimers) as an interacting substrate of Pak1. Pak1 directly phosphorylated TCoB in vitro and in vivo on serines 65 and 128 and colocalized with TCoB on newly polymerized microtubules and on centrosomes. TCoB interacted with the GTPase-binding domain of Pak1 and activated Pak1 in vitro and in vivo. In contrast to wild-type TCoB, an S65A, S128A double mutant and knock-down of the endogenous TCoB or Pak1 reduced microtubule polymerization, suggesting that Pak1 phosphorylation is necessary for normal TCoB function. Overexpression of TCoB dramatically increased the number of gamma tubulin-containing microtubule-organizing centers, a phenotype reminiscent of cells overexpressing Pak1. TCoB was overexpressed and phosphorylated in breast tumors. These findings reveal a novel role for TCoB and Pak1 in regulating microtubule dynamics. PMID- 15831478 TI - Direct p53 transcriptional repression: in vivo analysis of CCAAT-containing G2/M promoters. AB - In response to DNA damage, p53 activates G(1)/S blocking and apoptotic genes through sequence-specific binding. p53 also represses genes with no target site, such as those for Cdc2 and cyclin B, key regulators of the G(2)/M transition. Like most G(2)/M promoters, they rely on multiple CCAAT boxes activated by NF-Y, whose binding to DNA is temporally regulated during the cell cycle. NF-Y associates with p53 in vitro and in vivo through the alphaC helix of NF-YC (a subunit of NF-Y) and a region close to the tetramerization domain of p53. Chromatin immunoprecipitation experiments indicated that p53 is associated with cyclin B2, CDC25C, and Cdc2 promoters in vivo before and after DNA damage, requiring DNA-bound NF-Y. Following DNA damage, p53 is rapidly acetylated at K320 and K373 to K382, histones are deacetylated, and the release of PCAF and p300 correlates with the recruitment of histone deacetylases (HDACs)-HDAC1 before HDAC4 and HDAC5-and promoter repression. HDAC recruitment requires intact NF-Y binding sites. In transfection assays, PCAF represses cyclin B2, and a nonacetylated p53 mutant shows a complete loss of repression potential, despite its abilities to bind NF-Y and to be recruited on G(2)/M promoters. These data (i) detail a strategy of direct p53 repression through associations with multiple NF-Y trimers that is independent of sequence-specific binding of p53 and that requires C-terminal acetylation, (ii) suggest that p53 is a DNA damage sentinel of the G(2)/M transition, and (iii) delineate a new role for PCAF in cell cycle control. PMID- 15831479 TI - Cyclins D2 and D1 are essential for postnatal pancreatic beta-cell growth. AB - Regulation of adult beta-cell mass in pancreatic islets is essential to preserve sufficient insulin secretion in order to appropriately regulate glucose homeostasis. In many tissues mitogens influence development by stimulating D-type cyclins (D1, D2, or D3) and activating cyclin-dependent kinases (CDK4 or CDK6), which results in progression through the G(1) phase of the cell cycle. Here we show that cyclins D2 and D1 are essential for normal postnatal islet growth. In adult murine islets basal cyclin D2 mRNA expression was easily detected, while cyclin D1 was expressed at lower levels and cyclin D3 was nearly undetectable. Prenatal islet development occurred normally in cyclin D2(-/-) or cyclin D1(+/-) D2(-/-) mice. However, beta-cell proliferation, adult mass, and glucose tolerance were decreased in adult cyclin D2(-/-) mice, causing glucose intolerance that progressed to diabetes by 12 months of age. Although cyclin D1(+/-) mice never developed diabetes, life-threatening diabetes developed in 3-month-old cyclin D1( /+) D2(-/-) mice as beta-cell mass decreased after birth. Thus, cyclins D2 and D1 were essential for beta-cell expansion in adult mice. Strategies to tightly regulate D-type cyclin activity in beta cells could prevent or cure diabetes. PMID- 15831480 TI - Interaction of paxillin with poly(A)-binding protein 1 and its role in focal adhesion turnover and cell migration. AB - We have previously identified poly(A)-binding protein 1 (PABP1) as a ligand for paxillin and shown that the paxillin-PABP1 complex undergoes nucleocytoplasmic shuttling. By targeting the paxillin-binding subdomain sequences in PABP1, we have generated mutants of PABP1 that do not bind to cellular paxillin. Here we report that paxillin association is necessary for efficient nuclear export of PABP1 and that RNA interference of paxillin drives the nuclear accumulation of PABP1. Furthermore, ablation of paxillin-PABP1 association impeded a number of indices of cell motility including spreading on fibronectin, cell migration on two-dimensional matrices, and transmigration in Boyden chambers. These data indicate that PABP1 must associate with paxillin in order to be efficiently transported from the nucleus to the cytoplasm and that this event is necessary for cells to remodel their focal adhesions during cell migration. PMID- 15831481 TI - An evolutionarily conserved coiled-coil protein implicated in polycystic kidney disease is involved in basal body duplication and flagellar biogenesis in Trypanosoma brucei. AB - Trypanosoma brucei is a flagellated protozoan with a highly polarized cellular structure. TbLRTP is a trypanosomal protein containing multiple SDS22-class leucine-rich repeats and a coiled-coil domain with high similarity to a mammalian testis-specific protein of unknown function. Homologues are present in a wide range of higher eukaryotes including zebra fish, where the gene product has been implicated in polycystic kidney disease. Western blot analysis and immunofluorescence with antibodies against recombinant TbLRTP indicate that the protein is expressed throughout the trypanosome life cycle and localizes to distal zones of the basal bodies. Overexpression and RNA interference demonstrate that TbLRTP is important for faithful basal body duplication and flagellum biogenesis. Expression of excess TbLRTP suppresses new flagellum assembly, while reduction of TbLRTP protein levels often results in the biogenesis of additional flagellar axonemes and paraflagellar rods that, most remarkably, are intracellular and fully contained within the cytoplasm. The mutant flagella are devoid of membrane and are often associated with four microtubules in an arrangement similar to that observed in the normal flagellar attachment zone. Aberrant basal body and flagellar biogenesis in TbLRTP mutants also influences cell size and cytokinesis. These findings demonstrate that TbLRTP suppresses basal body replication and subsequent flagellar biogenesis and indicate a critical role for the LRTP family of proteins in the control of the cell cycle. These data further underscore the role of aberrant flagellar biogenesis as a disease mechanism. PMID- 15831482 TI - Selective regulation of c-jun gene expression by mitogen-activated protein kinases via the 12-o-tetradecanoylphorbol-13-acetate- responsive element and myocyte enhancer factor 2 binding sites. AB - To further understand how the mitogen-activated protein kinase (MAPK) signaling pathways regulate AP-1 activity, we have elucidated the physiological role of these cascades in the regulation of c-jun gene expression. c-Jun is a crucial component of AP-1 complexes and has been shown in vitro to be a point of integration of numerous signals that can differentially affect its expression as well as its transcriptional activity. Our strategy was based on the use of (i) genetically modified fibroblasts deficient in components of the MAPK cascades and (ii) pharmacological reagents. The results demonstrate that c-Jun NH(2)-terminal protein kinase (JNK) is essential for a basal level of c-Jun expression and for c Jun phosphorylation in response to stress. In addition to JNK, p38 MAPK or ERK1/2 and ERK5 are required for mediating UV radiation- or epidermal growth factor (EGF)-induced c-Jun expression, respectively. Further studies indicate that p38 MAPK inhibits the activation of JNK in response to EGF, causing a down-regulation of c-Jun. Overall, these data provide important insights into the mechanisms that ultimately determine the function of c-Jun as a regulator of cell fate. PMID- 15831483 TI - Small interfering RNAs that trigger posttranscriptional gene silencing are not required for the histone H3 Lys9 methylation necessary for transgenic tandem repeat stabilization in Neurospora crassa. AB - In Neurospora crassa, the introduction of a transgene can lead to small interfering RNA (siRNA)-mediated posttranscriptional gene silencing (PTGS) of homologous genes. siRNAs can also guide locus-specific methylation of Lys9 of histone H3 (Lys9H3) in Schizosaccharomyces pombe. Here we tested the hypothesis that transgenically derived siRNAs may contemporaneously both activate the PTGS mechanism and induce chromatin modifications at the transgene and the homologous endogenous gene. We carried out chromatin immunoprecipitation using a previously characterized albino-1 (al-1) silenced strain but detected no alterations in the pattern of histone modifications at the endogenous al-1 locus, suggesting that siRNAs produced from the transgenic locus do not trigger modifications in trans of those histones tested. Instead, we found that the transgenic locus was hypermethylated at Lys9H3 in our silenced strain and remained hypermethylated in the quelling defective mutants (qde), further demonstrating that the PTGS machinery is dispensable for Lys9H3 methylation. However, we found that a mutant in the histone Lys9H3 methyltransferase dim-5 was unable to maintain PTGS, with transgenic copies being rapidly lost, resulting in reversion of the silenced phenotype. These results indicate that the defect in PTGS of the Deltadim-5 strain is due to the inability to maintain the transgene in tandem, suggesting a role for DIM-5 in stabilizing such repeated sequences. We conclude that in Neurospora, siRNAs produced from the transgenic locus are used in the RNA-induced silencing complex-mediated PTGS pathway and do not communicate with an RNAi induced initiation of transcriptional gene silencing complex to effect chromatin based silencing. PMID- 15831484 TI - Defining the order in which Nmd3p and Rpl10p load onto nascent 60S ribosomal subunits. AB - The large ribosomal subunit protein Rpl10p is required for subunit joining and 60S export in yeast. We have recently shown that Rpl10p as well as the cytoplasmic GTPase Lsg1p are required for releasing the 60S nuclear export adapter Nmd3p from subunits in the cytoplasm. Here, we more directly address the order of Nmd3p and Rpl10p recruitment to the subunit. We show that Nmd3p can bind subunits in the absence of Rpl10p. In addition, we examined the basis of the previously reported dominant negative growth phenotype caused by overexpression of C-terminally truncated Rpl10p and found that these Rpl10p fragments are not incorporated into subunits in the nucleus but instead sequester the WD-repeat protein Sqt1p. Sqt1p is an Rpl10p binding protein that is proposed to facilitate loading of Rpl10p into the 60S subunit. Although Sqt1p normally only transiently binds 60S subunits, the levels of Sqt1p that can be coimmunoprecipitated by the 60S-associated GTPase Lsg1p are significantly increased by a dominant mutation in the Walker A motif of Lsg1p. This mutant Lsg1 protein also leads to increased levels of Sqt1p in complexes that are coimmunoprecipitated with Nmd3p. Furthermore, the dominant LSG1 mutant also traps a mutant Rpl10 protein that does not normally bind stably to the subunit. These results support the idea that Sqt1p loads Rpl10p onto the Nmd3p-bound subunit after export to the cytoplasm and that Rpl10p loading involves the GTPase Lsg1p. PMID- 15831485 TI - The human stress-activated protein kin17 belongs to the multiprotein DNA replication complex and associates in vivo with mammalian replication origins. AB - The human stress-activated protein kin17 accumulates in the nuclei of proliferating cells with predominant colocalization with sites of active DNA replication. The distribution of kin17 protein is in equilibrium between chromatin-DNA and the nuclear matrix. An increased association with nonchromatin nuclear structure is observed in S-phase cells. We demonstrated here that kin17 protein strongly associates in vivo with DNA fragments containing replication origins in both human HeLa and monkey CV-1 cells. This association was 10-fold higher than that observed with nonorigin control DNA fragments in exponentially growing cells. In addition, the association of kin17 protein to DNA fragments containing replication origins was also analyzed as a function of the cell cycle. High binding of kin17 protein was found at the G(1)/S border and throughout the S phase and was negligible in both G(0) and M phases. Specific monoclonal antibodies against kin17 protein induced a threefold inhibition of in vitro DNA replication of a plasmid containing a minimal replication origin that could be partially restored by the addition of recombinant kin17 protein. Immunoelectron microscopy confirmed the colocalization of kin17 protein with replication proteins like RPA, PCNA, and DNA polymerase alpha. A two-step chromatographic fractionation of nuclear extracts from HeLa cells revealed that kin17 protein localized in vivo in distinct protein complexes of high molecular weight. We found that kin17 protein purified within an approximately 600-kDa protein complex able to support in vitro DNA replication by means of two different biochemical methods designed to isolate replication complexes. In addition, the reduced in vitro DNA replication activity of the multiprotein replication complex after immunodepletion for kin17 protein highlighted for a direct role in DNA replication at the origins. PMID- 15831486 TI - Dok-R mediates attenuation of epidermal growth factor-dependent mitogen-activated protein kinase and Akt activation through processive recruitment of c-Src and Csk. AB - Dok-R has previously been shown to associate with the epidermal growth factor receptor (EGFR) and become tyrosine phosphorylated in response to EGF stimulation. The recruitment of Dok-R to the EGFR, which is mediated through its phosphotyrosine binding (PTB) domain, results in attenuation of mitogen-activated protein kinase (MAPK) activation. Dok-R's ability to attenuate EGF-driven MAPK activation is independent of its ability to recruit rasGAP, a known attenuator of MAPK activity, suggesting an alternate Dok-R-mediated pathway. Herein, we have determined the structural determinants within Dok-R that are required for its ability to attenuate EGF signaling and to associate with c-Src and with the Src family kinase (SFK)-inhibitory kinase, Csk. We demonstrate that Dok-R associates constitutively with c-Src through an SH3-dependent interaction and that this association is essential to Dok-R's ability to attenuate c-Src activity and diminish MAPK and Akt/PKB activity. We further illustrate that EGF-dependent phosphorylation of Dok-R requires SFK activity and, more specifically, that SFK dependent phosphorylation of tyrosine 402 on Dok-R facilitates the inducible recruitment of Csk. We propose that recruitment of Csk to Dok-R serves to bring Csk to c-Src and down-regulate its activity, resulting in a concomitant attenuation of MAPK and Akt/PKB activity. Furthermore, we demonstrate that Dok-R can abrogate c-Src's ability to protect the breast cancer cell line SKBR3 from anoikis and that an association with c-Src and Csk is required for this activity. Collectively these results demonstrate that Dok-R acts as an EGFR-recruited scaffolding molecule that processively assembles c-Src and Csk to attenuate signaling from the EGFR. PMID- 15831487 TI - Proteasomal ATPase-associated factor 1 negatively regulates proteasome activity by interacting with proteasomal ATPases. AB - The 26S proteasome, composed of the 20S core and the 19S regulatory complex, plays a central role in ubiquitin-dependent proteolysis by catalyzing degradation of polyubiquitinated proteins. In a search for proteins involved in regulation of the proteasome, we affinity purified the 19S regulatory complex from HeLa cells and identified a novel protein of 43 kDa in size as an associated protein. Immunoprecipitation analyses suggested that this protein specifically interacted with the proteasomal ATPases. Hence the protein was named proteasomal ATPase associated factor 1 (PAAF1). Immunoaffinity purification of PAAF1 confirmed its interaction with the 19S regulatory complex and further showed that the 19S regulatory complex bound with PAAF1 was not stably associated with the 20S core. Overexpression of PAAF1 in HeLa cells decreased the level of the 20S core associated with the 19S complex in a dose-dependent fashion, suggesting that PAAF1 binding to proteasomal ATPases inhibited the assembly of the 26S proteasome. Proteasomal degradation assays using reporters based on green fluorescent protein revealed that overexpression of PAAF1 inhibited the proteasome activity in vivo. Furthermore, the suppression of PAAF1 expression that is mediated by small inhibitory RNA enhanced the proteasome activity. These results suggest that PAAF1 functions as a negative regulator of the proteasome by controlling the assembly/disassembly of the proteasome. PMID- 15831488 TI - Niemann-Pick type C disease and intracellular cholesterol trafficking. PMID- 15831489 TI - Critical assessment of important regions in the subunit association and catalytic action of the severe acute respiratory syndrome coronavirus main protease. AB - The severe acute respiratory syndrome (SARS) coronavirus (CoV) main protease represents an attractive target for the development of novel anti-SARS agents. The tertiary structure of the protease consists of two distinct folds. One is the N-terminal chymotrypsin-like fold that consists of two structural domains and constitutes the catalytic machinery; the other is the C-terminal helical domain, which has an unclear function and is not found in other RNA virus main proteases. To understand the functional roles of the two structural parts of the SARS-CoV main protease, we generated the full-length of this enzyme as well as several terminally truncated forms, different from each other only by the number of amino acid residues at the C- or N-terminal regions. The quaternary structure and K(d) value of the protease were analyzed by analytical ultracentrifugation. The results showed that the N-terminal 1-3 amino acid-truncated protease maintains 76% of enzyme activity and that the major form is a dimer, as in the wild type. However, the amino acids 1-4-truncated protease showed the major form to be a monomer and had little enzyme activity. As a result, the fourth amino acid seemed to have a powerful effect on the quaternary structure and activity of this protease. The last C-terminal helically truncated protease also exhibited a greater tendency to form monomer and showed little activity. We concluded that both the C- and the N-terminal regions influence the dimerization and enzyme activity of the SARS-CoV main protease. PMID- 15831490 TI - Two-step mechanism that determines the donor binding specificity of human UDP-N acetylhexosaminyltransferase. AB - In its x-ray crystal structures, alpha-1,4-N-acteylhexosaminyltransferase (exostosin-like protein 2 (EXTL2)) forms no direct interaction with the N-acetyl group of the UDP-N-acetylhexosamine. Mutation of the residues that interact with the hydroxyl groups of the donor not only failed to abrogate donor binding but in fact increased binding affinity. Isothermal titration calorimetry is now used to examine the binding nature of various UDP-sugars in H2O and D2O solutions. UDP-N acetylhexosamines bind to EXTL2 with a high affinity in both solutions, resulting in a relatively large increase of entropy, whereas the weak binding of UDP galactose and -glucose, which occurred only in D2O solution, only slightly increased entropy. Thus, specific donor binding appears to undergo two distinct steps, beginning with the N-acetyl group expelling water from the donor. enzyme complex into the bulk solvent followed by positioning of the donor into the binding site for the subsequent interactions with the enzyme. PMID- 15831491 TI - Small GTPase proteins Rin and Rit Bind to PAR6 GTP-dependently and regulate cell transformation. AB - The novel small GTPases Rin and Rit are close relatives of Ras, and recent studies show that they play a role in mediating neuronal differentiation. However, the direct effectors of Rin and Rit have yet to be fully characterized. Here we showed that Rin and Rit directly bind to the PDZ domain of PAR6, a cell polarity-regulating protein, in a GTP-dependent manner both in vivo and in vitro. Moreover, Rin and Rit can form a ternary complex consisting of PAR6 and Rac/Cdc42, members of the Rho family of small GTPases modulating cell growth and polarity. This ternary complex synergistically potentiates cell transformation in NIH3T3 cells, and the interaction between Rin/Rit and the PDZ domain of PAR6 is important for this effect. These results suggest that the Rin/Rit-PAR6-Rac/Cdc42 ternary complex may work physiologically in the cells, such as in tumorigenesis. PMID- 15831492 TI - Ras-mediated loss of the pro-apoptotic response protein Par-4 is mediated by DNA hypermethylation through Raf-independent and Raf-dependent signaling cascades in epithelial cells. AB - The apoptosis-promoting protein Par-4 has been shown to be down-regulated in Ras transformed NIH 3T3 fibroblasts through the Raf/MEK/ERK MAPK pathway. Because mutations of the ras gene are most often found in tumors of epithelial origin, we explored the signaling pathways utilized by oncogenic Ras to down-regulate Par-4 in RIE-1 and rat ovarian surface epithelial (ROSE) cells. We determined that constitutive activation of the Raf, phosphatidylinositol 3-kinase, or Ral guanine nucleotide exchange factor effector pathway alone was not sufficient to down regulate Par-4 in RIE-1 or ROSE cells. However, treatment of Ras-transformed RIE 1 or ROSE cells with the MEK inhibitors U0126 and PD98059 increased Par-4 protein expression. Thus, although oncogenic Ras utilizes the Raf/MEK/ERK pathway to down regulate Par-4 in both fibroblasts and epithelial cells, Ras activation of an additional signaling pathway(s) is required to achieve the same outcome in epithelial cells. Methylation-specific PCR showed that the par-4 promoter is methylated in Ras-transformed cells through a MEK-dependent pathway and that treatment with the DNA methyltransferase inhibitor azadeoxycytidine restored Par 4 mRNA transcript and protein levels, suggesting that the mechanism for Ras mediated down-regulation of Par-4 is by promoter methylation. Support for this possibility is provided by our observation that Ras transformation was associated with up-regulation of Dnmt1 and Dnmt3 DNA methyltransferase expression. Finally, ectopic Par-4 expression significantly reduced Ras-mediated growth in soft agar, but not morphological transformation, highlighting the importance of Par-4 down regulation in specific aspects of Ras-mediated transformation of epithelial cells. PMID- 15831493 TI - Subunits of the translocon interact with components of the oligosaccharyl transferase complex. AB - Following initiation of translocation across the membrane of the endoplasmic reticulum via the translocon, polypeptide chains are N-glycosylated by the oligosaccharyl transferase (OT) enzyme complex. Translocation and N-glycosylation are concurrent events and would be expected to require juxtaposition of the translocon and the OT complex. To determine whether any of the subunits of the OT complex and translocon mediate interactions between the two complexes, we initiated a systematic study in budding yeast using the split-ubiquitin approach. Interestingly, the OT subunit Stt3p was found to interact only with Sec61p, whereas another OT subunit, Ost4p, was found to interact with all three components of the translocon, Sec61p, Sbh1p, and Sss1p. The OT subunit Wbp1p was found to interact very strongly with Sec61p and Sbh1p and weakly with Sss1p. Other OT subunits, Ost1p, Ost2p, and Swp1p were found to interact with Sec61p and either Sbh1p or Sss1p. Ost3p exhibited a weak interaction with Sec61p and Sbh1p, whereas Ost5p and Ost6p interacted very weakly with Sec61p and failed to interact with Sbh1p or Sss1p. We were able to confirm these split-ubiquitin findings by a chemical cross-linking technique. Based on our findings using these two techniques, we conclude that the association of these two complexes is stabilized via multiple protein-protein contacts. Based on extrapolation of the structural parameters of the crystal structure of the prokaryotic Sec complex to the eukaryotic complex, we propose a working model to understand the organization of the translocon-OT supercomplex. PMID- 15831494 TI - Function of mammalian LKB1 and Ca2+/calmodulin-dependent protein kinase kinase alpha as Snf1-activating kinases in yeast. AB - The Snf1/AMP-activated protein kinase (AMPK) family is important for metabolic regulation in response to stress. In the yeast Saccharomyces cerevisiae, the Snf1 kinase cascade comprises three Snf1-activating kinases, Pak1, Tos3, and Elm1. The only established mammalian AMPK kinase is LKB1. We show that LKB1 functions heterologously in yeast. In pak1Delta tos3Delta elm1Delta cells, LKB1 activated Snf1 catalytic activity and conferred a Snf(+) growth phenotype. Coexpression of STRADalpha and MO25alpha, which form a complex with LKB1, enhanced LKB1 function. Thus, the Snf1/AMPK kinase cascade is functionally conserved between yeast and mammals. Ca(2+)/calmodulin-dependent kinase kinase (CaMKK) shows more sequence similarity to Pak1, Tos3, and Elm1 than does LKB1. When expressed in pak1Delta tos3Delta elm1Delta cells, CaMKKalpha activated Snf1 catalytic activity, restored the Snf(+) phenotype, and also phosphorylated the activation loop threonine of Snf1 in vitro. These findings indicate that CaMKKalpha is a functional member of the Snf1/AMPK kinase family and support CaMKKalpha as a likely candidate for an AMPK kinase in mammalian cells. Analysis of the function of these heterologous kinases in yeast provided insight into the regulation of Snf1. When activated by LKB1 or CaMKKalpha, Snf1 activity was significantly inhibited by glucose, suggesting that a mechanism independent of the activating kinases can mediate glucose signaling in yeast. Finally, this analysis provided evidence that Pak1 functions in another capacity, besides activating Snf1, to regulate the nuclear enrichment of Snf1 protein kinase in response to carbon stress. PMID- 15831495 TI - Synthesis and characterization of a triphenylphosphonium-conjugated peroxidase mimetic. Insights into the interaction of ebselen with mitochondria. AB - Mitochondrial production of peroxides is a critical event in both pathology and redox signaling. Consequently their selective degradation within mitochondria is of considerable interest. Here we have explored the interaction of the peroxidase mimetic ebselen with mitochondria. We were particularly interested in whether ebselen was activated by mitochondrial glutathione (GSH) and thioredoxin, in determining whether an ebselen moiety could be targeted to mitochondria by conjugating it to a lipophilic cation, and in exploring the nature of ebselen binding to mitochondrial proteins. To achieve these goals we synthesized 2-[4-(4 triphenylphosphoniobutoxy) phenyl]-1,2-benzisoselenazol)-3(2H)-one iodide (MitoPeroxidase), which contains an ebselen moiety covalently linked to a triphenylphosphonium (TPP) cation. The fixed positive charge of TPP facilitated mass spectrometric analysis, which showed that the ebselen moiety was reduced by GSH to the selenol form and that subsequent reaction with a peroxide reformed the ebselen moiety. MitoPeroxidase and ebselen were effective antioxidants that degraded phospholipid hydroperoxides, prevented lipid peroxidation, and protected mitochondria from oxidative damage. Both peroxidase mimetics required activation by mitochondrial GSH or thioredoxin to be effective antioxidants. Surprisingly, conjugation to the TPP cation led to only a slight increase in the uptake of ebselen by mitochondria due to covalent binding of the ebselen moiety to proteins. Using antiserum against the TPP moiety we visualized those proteins covalently attached to the ebselen moiety. This analysis indicated that much of the ebselen present within mitochondria is bound to protein thiols through reversible selenenylsulfide bonds. Both MitoPeroxidase and ebselen decreased apoptosis induced by oxidative stress, suggesting that they can decrease mitochondrial oxidative stress. This exploration has led to new insights into the behavior of peroxidase mimetics within mitochondria and to their use in investigating mitochondrial oxidative damage. PMID- 15831496 TI - The secretory pathway Ca2+/Mn2+-ATPase 2 is a Golgi-localized pump with high affinity for Ca2+ ions. AB - Accumulation of Ca(2+) into the Golgi apparatus is mediated by sarco(endo)plasmic reticulum Ca(2+)-ATPases (SERCAs) and by secretory pathway Ca(2+)-ATPases (SPCAs). Mammals and birds express in addition to the housekeeping SPCA1 (human gene name ATP2C1, cytogenetic position 3q22.1) a homologous SPCA2 isoform (human gene name ATP2C2, cytogenetic position 16q24.1). We show here that both genes present an identical exon/intron layout. We confirmed that hSPCA2 has the ability to transport Ca(2+), demonstrated its Mn(2+)-transporting activity, showed its Ca(2+)- and Mn(2+)-dependent phosphoprotein intermediate formation, and documented the insensitivity of these functional activities to thapsigargin inhibition. The mRNA encoding hSPCA2 showed a limited tissue expression pattern mainly confined to the gastrointestinal and respiratory tract, prostate, thyroid, salivary, and mammary glands. Immunocytochemical localization in human colon sections presented a typical apical juxtanuclear Golgi-like staining. The expression in COS-1 cells allowed the direct demonstration of (45)Ca(2+) (K(0.5) = 0.27 microm) or (54)Mn(2+) transport into an A23187-releasable compartment. PMID- 15831497 TI - EPIYA motif is a membrane-targeting signal of Helicobacter pylori virulence factor CagA in mammalian cells. AB - Helicobacter pylori contributes to the development of peptic ulcers and atrophic gastritis. Furthermore, H. pylori strains carrying the cagA gene are more virulent than cagA-negative strains and are associated with the development of gastric adenocarcinoma. The cagA gene product, CagA, is translocated into gastric epithelial cells and localizes to the inner surface of the plasma membrane, in which it undergoes tyrosine phosphorylation at the Glu-Pro-Ile-Tyr-Ala (EPIYA) motif. Tyrosine-phosphorylated CagA specifically binds to and activates Src homology 2-containing protein-tyrosine phosphatase-2 (SHP-2) at the membrane, thereby inducing an elongated cell shape termed the hummingbird phenotype. Accordingly, membrane tethering of CagA is an essential prerequisite for the pathogenic activity of CagA. We show here that membrane association of CagA requires the EPIYA-containing region but is independent of EPIYA tyrosine phosphorylation. We further show that specific deletion of the EPIYA motif abolishes the ability of CagA to associate with the membrane. Conversely, reintroduction of an EPIYA sequence into a CagA mutant that lacks the EPIYA containing region restores membrane association of CagA. Thus, the presence of a single EPIYA motif is necessary for the membrane localization of CagA. Our results indicate that the EPIYA motif has a dual function in membrane association and tyrosine phosphorylation, both of which are critically involved in the activity of CagA to deregulate intracellular signaling, and suggest that the EPIYA motif is a crucial therapeutic target of cagA-positive H. pylori infection. PMID- 15831498 TI - The balance between acetylation and deacetylation controls Smad7 stability. AB - Transforming growth factor beta (TGFbeta) regulates multiple cellular processes via activation of Smad signaling pathways. We have recently demonstrated that the inhibitory Smad7 interacts with the acetyl transferase p300 and that p300 acetylates Smad7 on two lysine residues. These lysine residues are critical for Smurf-mediated ubiquitination of Smad7, and acetylation protects Smad7 from TGFbeta-induced degradation. In this study we demonstrate that Smad7 interacts with specific histone deacetylases (HDACs) and that the same HDACs are able to deacetylate Smad7. The interaction with HDACs is dependent on the C-terminal MH2 domain of Smad7. In addition, HDAC1-mediated deacetylation of Smad7 decreases the stability of Smad7 by enhancing its ubiquitination. Thus, our results demonstrate that the degradation of Smad7 is regulated by the balance between acetylation, deacetylation and ubiquitination, indicating that this could be a general mechanism to regulate the stability of cellular proteins. PMID- 15831499 TI - Cellular and gene expression responses involved in the rapid growth inhibition of human cancer cells by RNA interference-mediated depletion of telomerase RNA. AB - Inhibition of the up-regulated telomerase activity in cancer cells has previously been shown to slow cell growth but only after prior telomere shortening. Previously, we have reported that, unexpectedly, a hairpin short interfering RNA specifically targeting human telomerase RNA rapidly inhibits the growth of human cancer cells independently of p53 or telomere length and without bulk telomere shortening (Li, S., Rosenberg, J. E., Donjacour, A. A., Botchkina, I. L., Hom, Y. K., Cunha, G. R., and Blackburn, E. H. (2004) Cancer Res. 64, 4833-4840). Here we have demonstrated that such telomerase RNA knockdown in cancer cells does not cause telomere uncapping but rather induces changes in the global gene expression profile indicative of a novel response pathway, which includes suppression of specific genes implicated in angiogenesis and metastasis, and is distinct from the expression profile changes induced by telomere-uncapping mutant template telomerase RNAs. These cellular responses to depleting telomerase in human cancer cells together suggest that cancer cells are "telomerase-addicted" and uncover functions of telomerase in tumor growth and progression in addition to telomere maintenance. PMID- 15831500 TI - LXRbeta is required for adipocyte growth, glucose homeostasis, and beta cell function. AB - Liver X receptors (LXR) alpha and beta are nuclear oxysterol receptors with established roles in cholesterol, lipid, and carbohydrate metabolism. Although LXRs have been extensively studied in liver and macrophages, the importance for development and metabolism of other tissues and cell types is not as well characterized. We demonstrate here that although LXRalpha and LXRbeta are not required for adipocyte development per se, LXRbeta is required for the increase in adipocyte size that normally occurs with aging and diet-induced obesity. Similar food intake and oxygen consumption in LXRbeta-/- mice suggests that reduced storage of lipid in adipose tissue is not due to altered energy balance. Despite reduced amounts of adipose tissue, LXRbeta-/- mice on a chow diet have insulin sensitivity and levels of adipocyte hormones similar to wild type mice. However, these mice are glucose-intolerant due to impaired glucose-induced insulin secretion. Lipid droplets in pancreatic islets may result from accumulation of cholesterol esters as analysis of islet gene expression reveals that LXRbeta is required for expression of the cholesterol transporters, ABCA1 and ABCG1. Our data establish novel roles for LXRbeta in adipocyte growth, glucose homeostasis, and beta cell function. PMID- 15831501 TI - Proteomic and functional analyses reveal a mitochondrial dysfunction in P301L tau transgenic mice. AB - Transgenic mice overexpressing the P301L mutant human tau protein exhibit an accumulation of hyperphosphorylated tau and develop neurofibrillary tangles. The consequences of tau pathology were investigated here by proteomics followed by functional analysis. Mainly metabolism-related proteins including mitochondrial respiratory chain complex components, antioxidant enzymes, and synaptic proteins were identified as modified in the proteome pattern of P301L tau mice. Significantly, the reduction in mitochondrial complex V levels in the P301L tau mice revealed using proteomics was also confirmed as decreased in human P301L FTDP-17 (frontotemporal dementia with parkinsonism linked to chromosome 17) brains. Functional analysis demonstrated a mitochondrial dysfunction in P301L tau mice together with reduced NADH-ubiquinone oxidoreductase activity and, with age, impaired mitochondrial respiration and ATP synthesis. Mitochondrial dys-function was associated with higher levels of reactive oxygen species in aged transgenic mice. Increased tau pathology as in aged homozygous P301L tau mice revealed modified lipid peroxidation levels and the up-regulation of antioxidant enzymes in response to oxidative stress. Furthermore, P301L tau mitochondria displayed increased vulnerability toward beta-amyloid (Abeta) peptide insult, suggesting a synergistic action of tau and Abeta pathology on the mitochondria. Taken together, we conclude that tau pathology involves a mitochondrial and oxidative stress disorder possibly distinct from that caused by Abeta. PMID- 15831502 TI - Differential kinetics of cell surface loss of von Willebrand factor and its propolypeptide after secretion from Weibel-Palade bodies in living human endothelial cells. AB - The time course for cell surface loss of von Willebrand factor (VWF) and the propolypeptide of VWF (proregion) following exocytosis of individual Weibel Palade bodies (WPBs) from single human endothelial cells was analyzed. Chimeras of enhanced green fluorescent protein (EGFP) and full-length pre-pro-VWF (VWF EGFP) or the VWF propolypeptide (proregion-EGFP) were made and expressed in human umbilical vein endothelial cells. Expression of VWF-EGFP or proregion-EGFP resulted in fluorescent rod-shaped organelles that recruited the WPB membrane markers P-selectin and CD63. The WPB secretagogue histamine evoked exocytosis of these fluorescent WPBs and extracellular release of VWF-EGFP or proregion-EGFP. Secreted VWF-EGFP formed distinctive extracellular patches of fluorescence that were labeled with an extracellular antibody to VWF. The half-time for dispersal of VWF-EGFP from extracellular patches was 323.5 +/- 146.2 s (+/-S.D., n = 20 WPBs). In contrast, secreted proregion-EGFP did not form extracellular patches but dispersed rapidly from its site of release. The half-time for dispersal of proregion-EGFP following WPB exocytosis was 2.98 +/- 1.88 s (+/-S.D., n = 32 WPBs). The slow rate of loss of VWF-EGFP is consistent with the adhesive nature of this protein for the endothelial membrane. The much faster rate of loss of proregion-EGFP indicates that this protein does not interact strongly with extracellular VWF or the endothelial membrane and consequently may not play an adhesive role at the endothelial cell surface. PMID- 15831503 TI - Fertility and ageing. AB - The late 20th century trend to delay birth of the first child until the age at which female fecundity or reproductive capacity is lower has increased the incidence of age-related infertility. The trend and its consequences have also stimulated interest in the possible factors in the female and the male that may contribute to the decline in fecundity with age; in the means that exist to predict fecundity; and in the consequences for pregnancy and childbirth. In the female, the number of oocytes decreases with age until the menopause. Oocyte quality also diminishes, due in part to increased aneuploidy because of factors such as changes in spindle integrity. Although older male age affects the likelihood of conception, abnormalities in sperm chromosomes and in some components of the semen analysis are less important than the frequency of intercourse. Age is as accurate as any other predictor of conception with assisted reproductive technology. The decline in fecundity becomes clinically relevant when women reach their mid-30s, when even assisted reproduction treatment cannot compensate for the decline in fecundity associated with delaying attempts at conceiving. Pregnancies among women aged >40 years are associated with more non-severe complications, more premature births, more congenital malformations and more interventions at birth. PMID- 15831504 TI - Treatment option for sperm- or oocyte-related fertilization failure: assisted oocyte activation following diagnostic heterologous ICSI. AB - BACKGROUND: Failed fertilization occurs in 2-3% of ICSI cycles and is mainly due to lack of oocyte activation. Heterologous ICSI of patient's sperm in mouse oocytes allows discrimination between sperm- and oocyte-related aetiologies of activation failure. Assisted oocyte activation (AOA) by Ca-ionophore treatment can initiate fertilization in subsequent therapeutic ICSI. We report on diagnosis and clinical treatment in 17 patients with previously failed fertilization. METHODS: Sperm from patients were injected into mature mouse oocytes. Activation capacity was assessed by 2-cell formation (mouse oocyte activation test, MOAT). When no activation occurred, it was assumed that the spermatozoon was deficient; otherwise an oocyte-related factor was suspected. In a subsequent ICSI cycle, AOA was done by ICSI with CaCl2 followed by a Ca2+ ionophore exposure. Fertilization was checked 16-20 h later. Embryo transfer was on day 2 or 3. RESULTS: MOAT showed sperm-related activation deficiency in six globozoospermic patients and two patients with extreme oligoasthenoteratozoospermia. One patient with small sperm acrosomes had a normal activation percentage. In eight other patients, the MOAT revealed a relatively normal activation capacity of the sperm, indicating an oocyte-related defect. After AOA, fertilization rates were 77 and 71% in the sperm- and oocyte-related groups respectively. Five pregnancies were achieved in the globozoospermia group and three in cases of oocyte-related activation failure. CONCLUSIONS: Assisted oocyte activation enables normal fertilization and pregnancy in sperm- and oocyte-related fertilization failure. PMID- 15831505 TI - Post-mortem sperm retrieval in new European Union countries: case report. AB - This paper describes post-mortem sperm retrieval from a man who had given prior written consent to the retrieval and use of his sperm in assisted reproduction before his accidental death. The case illustrates some of the complex ethical and legal issues occurring in a new European Union (EU) country and the need for prior preparation by the medical teams involved. We also discuss a questionnaire sent to Centres of Assisted Reproduction in other new EU countries, which reveals a dearth of both legislation and guidelines for post-mortem sperm retrieval. If this method becomes more requested in these culturally diverse countries in the future, the situation demands public debate first of all at the national level. PMID- 15831506 TI - Multiple-birth risk associated with IVF and extended embryo culture: USA, 2001. AB - BACKGROUND: Multiple births are associated with serious adverse infant and maternal outcomes. The objective of this study was to assess the multiple-birth risk (MBR) associated with IVF and determine whether the risk is impacted by stage of embryo development at transfer. METHODS: A population-based sample of 50 819 IVF transfers utilizing day 3 or day 5 embryos performed in the USA in 2001 on women aged 20-40 years was used to assess MBR and live-birth rate (LBR), stratified by patient age, supernumerary embryo availability, and number of embryos transferred. RESULTS: Although significantly more day 5 than day 3 transfers used < or =2 embryos (69.2 versus 27.7%), the former were not associated with decreased MBR. MBR was high when >1 embryo was transferred, irrespective of embryo development stage. LBR were generally maximized with 2 embryos transferred, and for some (day 5 transfers, patients aged 35-37 years) with one embryo. Electing to transfer a single day 5 embryo appeared efficacious for some patients: women aged 20-37 years with supernumerary embryos cryopreserved had LBR of 31.6-39.5%. CONCLUSIONS: MBR is high when > or =2 embryos are transferred. Single embryo transfer is the only way to prevent many multiple births and associated adverse health outcomes. PMID- 15831507 TI - Development of a novel electrophoretic system for the isolation of human spermatozoa. AB - BACKGROUND: Optimization of assisted conception outcomes involves the development of rapid, safe, effective techniques for the isolation of functional human spermatozoa free from significant DNA damage. In this study we describe a novel electrophoretic sperm isolation technique that achieves these objectives. METHODS: The separation system consisted of a cassette comprising two 400 mul chambers separated by a polycarbonate filter containing 5 micromol/l pores and bounded by a 15 kDa polyacrylamide membrane to allow the free circulation of buffer. Semen was introduced into one chamber, current applied (75 mA at variable voltage) and within seconds a purified suspension of spermatozoa could be collected from the adjacent chamber. These cells were assessed for their count, viability, motility, morphology and DNA integrity. RESULTS: The suspensions generated by the electrophoretic separation technique contained motile, viable, morphologically normal spermatozoa and exhibited low levels of DNA damage. Moreover, these cell suspensions were free from contaminating cells, including leukocytes. The technique was comparable to discontinuous gradient centrifugation except that it took a fraction of the time and generated cells with significantly less DNA damage. CONCLUSION: Electrophoretic separation represents a highly effective, novel approach for the isolation of spermatozoa for assisted conception purposes. PMID- 15831508 TI - Misoprostol versus curettage in women with early pregnancy failure: impact on women's health-related quality of life. A randomized controlled trial. AB - BACKGROUND: We aimed to compare patients' health-related quality of life after a misoprostol strategy to a curettage in women with early pregnancy failure after failed expectant management. METHODS: A multicentre randomized clinical trial was performed in The Netherlands. In all, 154 women with early pregnancy failure confirmed at ultrasonography who had been managed expectantly unsuccessfully for > or =1 week were randomly assigned to undergo either treatment with misoprostol (n=79) or curettage (n=75). The main outcome measures were health-related quality of life and satisfaction with treatment. RESULTS: In the misoprostol strategy 47% of the women needed additional curettage, as compared to 4% after curettage. In both groups, health-related quality of life was impaired most severely 2 days after treatment. In the misoprostol group, health-related quality of life was more severely impaired; after 2 days this was due to more pain and after 2 and 6 weeks this was due to a worse general health perception. Health-related quality of life was temporarily significantly more impaired in women in whom misoprostol failed as compared to women in whom misoprostol treatment was successful. In both treatment groups, an equal percentage of women (58%) would choose the same treatment in the future. In women treated with misoprostol, however, this choice depended on the initial success of misoprostol: in cases where misoprostol had caused complete evacuation, 76% of the women would opt for the same treatment, whereas only 38% of women who needed curettage after unsuccessful misoprostol would do so (P<0.01). CONCLUSION: Our study shows that, although both the misoprostol strategy and the curettage strategy resulted in complete evacuation in the end, women are willing to accept some disadvantages of misoprostol to avoid curettage. A treatment inconvenience using misoprostol is accepted as long as initial evacuation rate is high. This finding should be an integral part of counselling women when deciding upon management of early pregnancy failure. PMID- 15831509 TI - Development of an experimental model of endometriosis using mice that ubiquitously express green fluorescent protein. AB - BACKGROUND: Aiming at improving an animal model of endometriosis, we developed a homologous mouse model using 'green mice' that ubiquitously express green fluorescent protein. METHODS: Endometrial fragment obtained from estradiol (E2) supplemented ovariectomized 'green mice' was minced and injected into the peritoneal cavity of ovariectomized wild-type mice. The recipient wild mice were raised with or without E2 supplementation for 2 weeks, and then were euthanized. Endometriotic lesions that developed in the abdomen were examined both macroscopically and microscopically under fluorescence, and weight of the lesions was measured. RESULTS: The endometriotic lesions were more clearly detected under fluorescence imaging than by conventional macroscopic examination. Histologically, endometriotic lesions deriving from 'green mice' were sharply distinguished from surrounding host tissues under fluorescence microscopy. More lesions developed in E2-supplemented than control recipient mice. The measured fluorescence intensity of endometriotic lesions showed significant positive correlation with their weight (R=0.844, P<0.0001), and was significantly higher in E2-supplemented mice than in vehicle-supplemented mice (P=0.0062). CONCLUSION: The present endometriosis model using 'green mice' would be useful for expeditious identification and quantitative evaluation of endometriotic lesions. PMID- 15831510 TI - A trial to restore defective human sperm centrosomal function. AB - BACKGROUND: In human fertilization, sperm centrosome function is essential for male and female pronuclear movement and fusion. In this study, we investigated the possibility of restoring human sperm centrosomal function in sperm exhibiting abnormalities in microtubule organization. METHODS: Semen was obtained from both a fertile donor and a patient with dysplasia of the fibrous sheath (DFS). Following heterologous ICSI using human sperm, we examined microtubules and chromatin configuration in bovine oocytes. Sperm were treated with dithiothreitol (DTT) prior to ICSI, while the oocytes were treated with the cytoskeletal stabilizer paclitaxel after ICSI. RESULTS: The combination of DTT and paclitaxel treatment induced microtubule organization in dead sperm from the fertile donor following heterologous ICSI. This treatment, however, was not effective for DFS sperm. In addition, expression of centrin, a protein functioning within the sperm centrosome, was reduced in DFS sperm from that of the normal levels observed in fertile donor sperm. CONCLUSION: These results indicate that sperm centrosomal function could be induced by the treatment of sperm with DTT before ICSI and of oocytes with paclitaxel after ICSI. DFS sperm are likely to exhibit such severe dysfunction of sperm centrosome that cannot be compensated for by this treatment; therefore, this method may be a practical way to discern the degree of sperm centrosomal dysfunction. PMID- 15831511 TI - Estrogen and progesterone receptor expression in macrophages and regulation of hepatocyte growth factor by ovarian steroids in women with endometriosis. AB - BACKGROUND: Information regarding macrophage-mediated regulation of hepatocyte growth factor (HGF) by ovarian steroid hormones in women with endometriosis is limited. Therefore, we investigated the regulation of HGF by steroid hormones in isolated macrophages and stromal cells derived from women with or without endometriosis. METHODS: We isolated CD68 immunoreactive adherent macrophages in vitro from 46 women with endometriosis and 30 women without endometriosis. Estrogen receptor (ER) and progesterone receptor (PR) expression in macrophages was demonstrated by immunohistochemistry and RT-PCR. Production of HGF in the culture media of basal and ovarian steroid-stimulated macrophages was examined by enzyme-linked immunosorbent assay. Expression of mRNA for HGF and its receptor, c Met in macrophages and stromal cells in response to ovarian steroid was investigated by RT-PCR. The single and combined effect of HGF and estrogen on the growth of macrophages and stromal cells was analysed by bromodeoxyuridine (BrdU) incorporation. RESULTS: ER and PR were expressed in isolated macrophages and intact tissue at the protein and mRNA levels. Macrophages derived from women with endometriosis produced significantly higher concentration of HGF (352.2 +/- 4.9 pg/ml) in conditioned media after treatment with estradiol (10(-8) mol/l) than that of basal macrophages (221.5 +/- 32.8 pg/ml, P<0.05) or women without endometriosis (170.6 +/- 2.6 pg/ml, P<0.05). These effects were less evident after treatment with progesterone. Treatment with tamoxifen (10(-6) mol/l) reversed the production of HGF and other macromolecules. Secretion of HGF in response to ovarian steroids was further enhanced after activation with lipopolysaccharide. The mRNA expressions of HGF and its receptor, c-Met, were also detected in macrophages and stroma in response to estrogen, suggesting an autocrine regulation. HGF mRNA expression was higher in cells of women with endometriosis than non-endometriosis women. Bromodeoxyuridine incorporation indicated that exogenous stimulation with HGF and estrogen, either alone or in combination, significantly increased the cell proliferation of both endometrial stroma and macrophages compared to that of non-endometriosis or non-treated cells. CONCLUSION: These results suggest that besides other inflammatory mediators, ovarian steroids also participate in the production of HGF by peritoneal macrophages which may be involved in the growth of endometriosis either alone or in combination with estrogen. PMID- 15831512 TI - Association between single nucleotide polymorphisms of estrogen receptor alpha gene and efficacy of HRT on bone mineral density in post-menopausal Japanese women. AB - BACKGROUND: Although HRT for post-menopausal women can protect against bone loss, variations in bone responses exist. We studied whether single nucleotide polymorphisms (SNP) of the estrogen receptor-alpha (ERalpha) gene contribute to the effect of HRT on lumbar spine bone mineral density (BMD). METHODS: Subjects were 84 post-menopausal women who had been taking HRT for 3 years to treat osteopenia or osteoporosis. Eighteen SNP in the ERalpha gene were characterized by a single nucleotide primer extension assay. RESULTS: Genotyping of the 84 individuals revealed that all SNP were quite common, the minor allele frequency being > or = 20%. A SNP in intron 6 (IVS6+14144) was significantly associated with the response to HRT for the first 3 years after starting treatment (P = 0.043, 0.025 and 0.032 for the first, second and third years respectively). Haplotype analysis revealed that a combination of SNP IVS6+14144 and IVS4+4238 was significantly correlated with the response to HRT; women with haplotype G-G (IVS6 14144-IVS4 4238) showed a significantly higher response (P = 0.014, 0.043 and 0.010 for the first second and third year respectively). CONCLUSIONS: These results suggest that a specific SNP and the haplotype of the selected SNP could be used to predict the effect of HRT on lumbar BMD. PMID- 15831513 TI - Intra-cervical versus i.v. fentanyl for abortion. AB - BACKGROUND: The majority of abortions are performed using a para-cervical block (without general anaesthesia) and involve a significant amount of pain. If fentanyl was given with the lidocaine in the para-cervical block, it potentially could improve pain control while decreasing side effects and avoiding i.v. access for women having abortions. METHODS: This was a randomized double-blind placebo controlled trial of two treatment arms: (i) para-cervical block with 100 microg of fentanyl i.v; or (ii) para-cervical block with 100 microg of fentanyl intra cervically (i.c.) for first trimester abortion. The setting was a free-standing urban abortion clinic. The outcome measures were pain scores and side effects. RESULTS: A total of 104 women received the fentanyl i.v. and 98 received the fentanyl i.c. The two groups were similar with respect to age, gestational age, obstetric history, anxiety and depression. Pain scores (0-10) were 4.7 and 5.7 for dilation (P = 0.01) and 3.8 and 5.6 for suctioning (P < 0.001) in the i.v. and i.c. groups, respectively. Side effects were similar, but more women in the i.v. group received anti-emetics. More women in the i.c. group were dissatisfied with the pain control. CONCLUSION: I.v. fentanyl is more effective than i.c. fentanyl for pain control in abortion. PMID- 15831514 TI - Predicting ongoing pregnancy following ovulation induction with recombinant FSH in women with polycystic ovary syndrome. AB - BACKGROUND: Ovulation induction with recombinant FSH (rFSH) is common in women with polycystic ovary syndrome (PCOS) not responding to clomiphene citrate treatment, despite the associated risk of multiple pregnancies. We analysed clinical, ultrasonographic and endocrine parameters during initial screening of women with clomiphene citrate-resistant PCOS as predictors of ongoing pregnancy within 12 months of treatment following ovulation induction with rFSH. METHODS: Eighty-five women were allocated to receive rFSH as part of a multicentre clinical trial. rFSH was administered in a chronic low-dose step-up protocol. The primary end-point was an ongoing pregnancy within 12 months. A logistic model was built using clinical, ultrasonographic and endocrine parameters to predict the response to rFSH treatment, adjusted for the number of cycles performed. RESULTS: In total, 85 women underwent 272 treatment cycles with rFSH, of which 57 women (67%) achieved an ongoing pregnancy. Oligomenorrhoea, shorter duration of infertility and a lower free androgen index (FAI) were associated with higher chances of an ongoing pregnancy, resulting in a predictive model with a modest discriminative power (area under the curve 0.72, 95% confidence interval 0.64 0.79) that allowed us to distinguish between women with a probability of <5% of attaining an ongoing pregnancy and women with a probability of >25% of doing so. CONCLUSION: A model consisting of oligo/amenorrhoea, duration of infertility and FAI level allowed a distinction to be made between women with a poor chance and women with a good chance of achieving an ongoing pregnancy. PMID- 15831515 TI - Serum fluctuations of total and free tryptophan levels during the menstrual cycle are related to gonadotrophins and reflect brain serotonin utilization. AB - BACKGROUND: Serotoninergic (5-HT) neurons are suggested to regulate estrous cycle in animal models. In the present study we evaluated whether a relationship exists between the serotoninergic precursors in the central nervous system and the gonadotrophin-ovarian cyclic function. METHODS: We measured 5-HT precursors [free (FT) and total (TT) tryptophan] and LH, FSH and 17beta-estradiol (E2) levels in the serum of 15 fertile women with normal menstrual cycles during the follicular (cycle days 1-5, 7-11), mid-cycle (cycle days 14-16) and luteal (cycle days 17 19, 22-24) phases. RESULTS: TT and FT were significantly increased in the 7-11 and 17-19 cycle days and were decreased at mid-cycle (P < 0.01), with a cyclic and opposite behaviour when compared to that of FSH and LH. Indeed, correlation analysis through the matrix of mean values showed that LH was negatively correlated to TT (r = -0.636) and FT (r = -0.574), as well as FSH (TT, r = 0.655; FT, r = -0.663), and that TT and FT were positively correlated to each other (r = 0.801; P < 0.001). Furthermore, whilst the two FT peaks reached approximately the same levels in the follicular and luteal phase, TT levels were approximately 30% higher in the luteal than in the follicular phase of the cycle: thus in the first (follicular) phase FT peak was relatively higher than that of TT, whereas the contrary occurred in the second (luteal) phase of the cycle. CONCLUSIONS: Both TT and FT levels have cyclic variations throughout the menstrual cycle, being lowest at mid-cycle (14-16 cycle days), concomitant with the highest LH and FSH concentrations, and higher before and after mid-cycle phase, coinciding with the lowest circulating LH/FSH levels. Since TT and FT levels in the plasma have cyclic changes, our study: (i) suggests that a consumption of serum serotonin precursors takes place concomitant with gonadotrophin release during menstrual cycle; (ii) may represent an in vivo model to investigate this relationship in women in different physiopathological conditions. PMID- 15831516 TI - Functional genomics identifies a mechanism for estrogen activation of the retinoic acid receptor alpha1 gene in breast cancer cells. AB - The identification of estrogen receptor (ERalpha) target genes is crucial to our understanding of its predominant role in breast cancer. In this study, we used a chromatin immunoprecipitation (ChIP)-cloning strategy to identify ERalpha regulatory modules and associated target genes in the human breast cancer cell line MCF-7. We isolated 12 transcriptionally active genomic modules that recruit ERalpha and the coactivator steroid receptor coactivator (SRC)-3 to different intensities in vivo. One of the ERalpha-regulatory modules identified is located 3.7 kb downstream of the first transcriptional start site of the RARA locus, which encodes retinoic acid receptor alpha1 (RARalpha1). This module, which includes an estrogen response element (ERE), is conserved between the human and mouse genomes. Direct binding of ERalpha to the ERE was shown using EMSAs, and transient transfections in MCF-7 cells demonstrated that endogenous ERalpha can induce estrogen-dependent transcriptional activation from the module or the ERE linked to a heterologous promoter. Furthermore, ChIP assays showed that the coregulators SRC-1, SRC-3, and receptor-interacting protein 140 are recruited to this intronic module in an estrogen-dependent manner. As expected from previous studies, the transcription factor Sp1 can be detected at the RARA alpha1 promoter by ChIP. However, treatment with estradiol did not influence Sp1 recruitment nor help recruit ERalpha to the promoter. Finally, ablation of the intronic ERE was sufficient to abrogate the up-regulation of RARA alpha1 promoter activity by estradiol. Thus, this study uncovered a mechanism by which ERalpha significantly activates RARalpha1 expression in breast cancer cells and exemplifies the utility of functional genomics strategies in identifying long-distance regulatory modules for nuclear receptors. PMID- 15831517 TI - A peroxisome proliferator-activated receptor alpha/gamma dual agonist with a unique in vitro profile and potent glucose and lipid effects in rodent models of type 2 diabetes and dyslipidemia. AB - LSN862 is a novel peroxisome proliferator-activated receptor (PPAR)alpha/gamma dual agonist with a unique in vitro profile that shows improvements on glucose and lipid levels in rodent models of type 2 diabetes and dyslipidemia. Data from in vitro binding, cotransfection, and cofactor recruitment assays characterize LSN862 as a high-affinity PPARgamma partial agonist with relatively less but significant PPARalpha agonist activity. Using these same assays, rosiglitazone was characterized as a high-affinity PPARgamma full agonist with no PPARalpha activity. When administered to Zucker diabetic fatty rats, LSN862 displayed significant glucose and triglyceride lowering and a significantly greater increase in adiponectin levels compared with rosiglitazone. Expression of genes involved in metabolic pathways in the liver and in two fat depots from compound treated Zucker diabetic fatty rats was evaluated. Only LSN862 significantly elevated mRNA levels of pyruvate dehydrogenase kinase isozyme 4 and bifunctional enzyme in the liver and lipoprotein lipase in both fat depots. In contrast, both LSN862 and rosiglitazone decreased phosphoenol pyruvate carboxykinase in the liver and increased malic enzyme mRNA levels in the fat. In addition, LSN862 was examined in a second rodent model of type 2 diabetes, db/db mice. In this study, LSN862 demonstrated statistically better antidiabetic efficacy compared with rosiglitazone with an equivalent side effect profile. LSN862, rosiglitazone, and fenofibrate were each evaluated in the humanized apoA1 transgenic mouse. At the highest dose administered, LSN862 and fenofibrate reduced very low-density lipoprotein cholesterol, whereas, rosiglitazone increased very low-density lipoprotein cholesterol. LSN862, fenofibrate, and rosiglitazone produced maximal increases in high-density lipoprotein cholesterol of 65, 54, and 30%, respectively. These findings show that PPARgamma full agonist activity is not necessary to achieve potent and efficacious insulin-sensitizing benefits and demonstrate the therapeutic advantages of a PPARalpha/gamma dual agonist. PMID- 15831518 TI - Requirement of retinoic acid receptor isotypes alpha, beta, and gamma during the initial steps of neural differentiation of PCC7 cells. AB - Retinoic acid (RA) is indispensable for morphogenesis and differentiation of several tissues, including the nervous system. The requirement of the RA receptor (RAR) isotypes alpha, beta, and gamma and the putative role of retinoid X receptor-(RXR) signaling in RA-induced neural differentiation, was analyzed. For this compound-selective retinoids and the murine embryonal carcinoma cell line PCC7, a model system for RA-dependent neural differentiation was used. The present paper shows that proliferating PCC7 cells primarily express RXRalpha and RARalpha, lower levels of RXRbeta, and barely detectable amounts of RARbeta, RARgamma, and RXRgamma. At receptor-selective concentrations, only a RARalpha or RARgamma agonist induced the typical tissue-like differentiation pattern consisting of neuronal and nonneuronal cells. Differentiation-associated processes, such as the down-regulation of Oct4, up-regulation of certain nuclear receptors and proneuronal genes, and the induction of neuronal markers could be triggered by receptor-selective concentrations of a RARalpha-, beta-, or gamma selective agonist, although with distinct efficacy. The differences are only partially explained by the distinct RARalpha, beta, and gamma expression levels and the dissociation constants for the bound retinoids, suggesting differential requirement of RAR isotypes during the initial stages of neural differentiation of PCC7 cells. PMID- 15831519 TI - Multiple signaling pathways regulating steroidogenesis and steroidogenic acute regulatory protein expression: more complicated than we thought. AB - Steroid hormone biosynthesis in steroidogenic cells is regulated through trophic hormone activation of protein kinase A (PKA) signaling pathways. However, many examples of the regulation of steroid synthesis via pathways other than the PKA pathway have been documented. In some cases these pathways act independently of PKA activation whereas in other cases, they act synergistically with it. The current understanding of additional signaling pathways and factors, such as the protein kinase C pathway, arachidonic acid metabolites, growth factors, chloride ion, the calcium messenger system, and others capable of regulating/modulating steroid hormone biosynthesis, and in many cases steroidogenic acute regulatory protein expression, are discussed in this review. PMID- 15831520 TI - The modulator of nongenomic actions of the estrogen receptor (MNAR) regulates transcription-independent androgen receptor-mediated signaling: evidence that MNAR participates in G protein-regulated meiosis in Xenopus laevis oocytes. AB - Classical steroid receptors mediate many transcription-independent (nongenomic) steroid responses in vitro, including activation of Src and G proteins. Estrogen triggered activation of Src can be regulated by the modulator of nongenomic actions of the estrogen receptor (MNAR), which binds to estrogen receptors and Src to create a signaling complex. In contrast, the mechanisms regulating steroid induced G protein activation are not known, nor are the physiologic responses mediated by MNAR. These studies demonstrate that MNAR regulates the biologically relevant process of meiosis in Xenopus laevis oocytes. MNAR was located throughout oocytes, and reduction of its expression by RNA interference markedly enhanced testosterone-triggered maturation and activation of MAPK. Additionally, Xenopus MNAR augmented androgen receptor (AR)-mediated transcription in CV1 cells through activation of Src. MNAR and AR coimmunoprecipitated as a complex involving the LXXLL-rich segment of MNAR and the ligand binding domain of AR. MNAR and Gbeta also precipitated together, with the same region of MNAR being important for this interaction. Finally, reduction of MNAR expression decreased Gbetagamma-mediated signaling in oocytes. MNAR therefore appears to participate in maintaining meiotic arrest, perhaps by directly enhancing Gbetagamma-mediated inhibition of meiosis. Androgen binding to AR might then release this inhibition, allowing maturation to occur. Thus, MNAR may augment multiple nongenomic signals, depending upon the context and cell type in which it is expressed. PMID- 15831521 TI - Xenobiotic stress induces hepatomegaly and liver tumors via the nuclear receptor constitutive androstane receptor. AB - The constitutive androstane receptor (CAR, NR1I3) is a central regulator of xenobiotic metabolism. CAR activation induces hepatic expression of detoxification enzymes and transporters and increases liver size. Here we show that CAR-mediated hepatomegaly is a transient, adaptive response to acute xenobiotic stress. In contrast, chronic CAR activation results in hepatocarcinogenesis. In both acute and chronic xenobiotic responses, hepatocyte DNA replication is increased and apoptosis is decreased. These effects are absent in CAR null mice, which are completely resistant to tumorigenic effects of chronic xenobiotic stress. In the acute response, direct up-regulation of Mdm2 expression by CAR contributes to both increased DNA replication and inhibition of p53-mediated apoptosis. These results demonstrate an essential role for CAR in regulating both liver homeostasis and tumorigenesis in response to xenobiotic stresses, and they also identify a specific molecular mechanism linking chronic environmental stress and tumor formation. PMID- 15831522 TI - Thyroid hormone induces cardiac myocyte hypertrophy in a thyroid hormone receptor alpha1-specific manner that requires TAK1 and p38 mitogen-activated protein kinase. AB - Alterations in TR [thyroid hormone (TH) receptor]1 isoform expression have been reported in models of both physiologic and pathologic cardiac hypertrophy as well as in patients with heart failure. In this report, we demonstrate that TH induces hypertrophy as a direct result of binding to the TRalpha1 isoform and, moreover, that overexpression of TRalpha1 alone is also associated with a hypertrophic phenotype, even in the absence of ligand. The mechanism of TH and TRalpha1 specific hypertrophy is novel for a nuclear hormone receptor and involves the transforming growth factor beta-activated kinase (TAK1) and p38. Mitigating TRalpha1 effects, both TRalpha2 and TRbeta1 attenuate TRalpha1-induced myocardial growth and gene expression by diminishing TAK1 and p38 activities, respectively. These findings refine our previous observations on TR expression in the hypertrophied and failing heart and suggest that manipulation of thyroid hormone signaling in an isoform-specific manner may be a relevant therapeutic target for altering the pathologic myocardial program. PMID- 15831523 TI - Role of the basic helix-loop-helix transcription factor, scleraxis, in the regulation of Sertoli cell function and differentiation. AB - Sertoli cells are a postmitotic terminally differentiated cell population in the adult testis that form the seminiferous tubules and provide the microenvironment and structural support for developing germ cells. The transcription factors that regulate Sertoli cell differentiation remain to be elucidated. The basic helix loop-helix transcription factors are involved in the differentiation of a variety of cell lineages during development and are expressed in pubertal Sertoli cells. A yeast-two-hybrid procedure was used to screen a Sertoli cell library from 20-d old pubertal rats to identify dimerization partners with the ubiquitous E47 basic helix-loop-helix transcription factor. Scleraxis was identified as one of the interacting partners. Among the cell types of the testis, scleraxis expression was found to be specific to Sertoli cells. Analysis of the expression pattern of scleraxis mRNA in developing Sertoli cells revealed an increase in scleraxis message at the onset of puberty. Sertoli cells respond to FSH to promote expression of differentiated gene products such as transferrin that aid in proper development of the germ cells. Analysis of the hormonal regulation of scleraxis expression revealed a 4-fold increase in scleraxis mRNA in response to the presence of FSH or dibutryl cAMP in cultured Sertoli cells. An antisense oligonucleotide procedure and overexpression analysis were used to determine whether scleraxis regulates the expression of Sertoli cell differentiated gene products. An antisense oligonucleotide to scleraxis down-regulated transferrin promoter activity in Sertoli cells. A transient overexpression of scleraxis in Sertoli cells stimulated transferrin and androgen binding protein promoter activities and the expression of a number of differentiated genes. Observations suggest scleraxis functions in a number of adult tissues and is involved in the regulation and maintenance of Sertoli cell function and differentiation. This is one of the first adult and nontendon/chondrocyte-associated functions described for scleraxis. PMID- 15831524 TI - Distinctive actions of membrane-targeted versus nuclear localized estrogen receptors in breast cancer cells. AB - Estrogens regulate multiple activities in breast cancer cells, including proliferation. Whereas these hormones are most commonly known to regulate gene transcription through direct interaction with estrogen receptors (ERs) and with specific DNA sequences of target genes, recent studies show that ER also activates a number of rapid signaling events that are initiated at the cell membrane. To study the membrane-initiated effects of estrogen and separate them from the activities initiated by the nuclear localized ER in human breast cancer cells, we generated MDA-MB-231 breast cancer cell lines that have stably integrated either the wild-type nuclear form of ER (WT-ER) or a modified, membrane-targeted ER (MT-ER) that lacks a nuclear localization sequence and is dually acylated with a myristoylation sequence at the N terminus and a palmitoylation sequence at the C terminus. We demonstrate that MT-ER is membrane localized in the absence of estradiol (E2), showing punctate membrane and cytoplasmic speckles after E2 exposure. In contrast to WT-ER, MT-ER was not down regulated by E2 or by antiestrogen ICI 182,780 exposure, and MT-ER failed to regulate endogenous E2-responsive genes highly up-regulated by WT-ER. Cells expressing MT-ER showed a greater serum response element-mediated transcriptional response that was partially inhibited by antiestrogen ICI 182,780. The MT-ER and WT-ER differentially altered ERK1/2 and Akt activities and the proliferation of breast cancer cells in response to E2. Hence, this study reveals distinct actions of the MT-ER vs. the WT-ER in effecting estrogen actions in breast cancer cells. PMID- 15831525 TI - Physiological role for the cochaperone FKBP52 in androgen receptor signaling. AB - Molecular chaperones mediate multiple aspects of steroid receptor function, but the physiological importance of most receptor-associated cochaperones has not been determined. To help fill this gap, we targeted for disruption the mouse gene for the 52-kDa FK506 binding protein, FKBP52, a 90-kDa heat shock protein (Hsp90) binding immunophilin found in steroid receptor complexes. A mouse line lacking FKBP52 (52KO) was generated and characterized. Male 52KO mice have several defects in reproductive tissues consistent with androgen insensitivity; among these defects are ambiguous external genitalia and dysgenic prostate. FKBP52 and androgen receptor (AR) are coexpressed in prostate epithelial cells of wild-type mice. However, FKBP52 and AR are similarly coexpressed in testis even though testis morphology and spermatogenesis in 52KO males are usually normal. Molecular studies confirm that FKBP52 is a component of AR complexes, and cellular studies in yeast and human cell models demonstrate that FKBP52 can enhance AR-mediated transactivation. AR enhancement requires FKBP52 peptidylprolyl isomerase activity as well as Hsp90-binding ability, and enhancement probably relates to an affect of FKBP52 on AR-folding pathways. In the presence of FKBP52, but not other cochaperones, the function of a minimally active AR point mutant can be dramatically restored. We conclude that FKBP52 is an AR folding factor that has critically important physiological roles in some male reproductive tissues. PMID- 15831526 TI - Regulation of cytochrome b5 gene transcription by Sp3, GATA-6, and steroidogenic factor 1 in human adrenal NCI-H295A cells. AB - Sex steroid synthesis requires the 17,20 lyase activity of P450c17, which is enhanced by cytochrome b5, acting as an allosteric factor to promote association of P450c17 with its electron donor, P450 oxidoreductase. Cytochrome b5 is preferentially expressed in the fetal adrenal and postadrenarchal adrenal zona reticularis; the basis of this tissue-specific, developmentally regulated transcription of the b5 gene is unknown. We found b5 expression in all cell lines tested, including human adrenal NCI-H295A cells, where its mRNA is reduced by cAMP and phorbol ester. Multiple sites, between -83 and -122 bp upstream from the first ATG, initiate transcription. Deletional mutagenesis localized all detectable promoter activity within -327/+15, and deoxyribonuclease I footprinting identified protein binding at -72/-107 and -157/-197. DNA segments 65/-40, -114/-70 and -270/-245 fused to TK32/Luc yielded significant activity, and mutations in their Sp sites abolished that activity; electrophoretic mobility shift assay (EMSA) showed that Sp3, but not Sp1, binds to these Sp sites. Nuclear factor 1 (NF-1) and GATA-6, but not GATA-4 bind to the NF-1 and GATA sites in 157/-197. In Drosophila S2 cells, Sp3 increased -327/Luc activity 58-fold, but Sp1 and NF-1 isoforms were inactive. Mutating the three Sp sites ablated activity without or with cotransfection of Sp1/Sp3. In NCI-H295A cells, mutating the three Sp sites reduced activity to 39%; mutating the Sp, GATA, and NF-1 sites abolished activity. In JEG-3 cells, GATA-4 was inactive, GATA-6 augmented -327/Luc activity to 231% over the control, and steroidogenic factor 1 augmented activity to 655% over the control; these activities required the Sp and NF-1 sites. Transcription of cytochrome b5 shares many features with the regulation of P450c17, whose activity it enhances. PMID- 15831527 TI - Silibinin inhibits ultraviolet B radiation-induced mitogenic and survival signaling, and associated biological responses in SKH-1 mouse skin. AB - Ultraviolet B (UVB) radiation is a complete skin carcinogen causing DNA damage as a tumor-initiating event and activating signaling cascades that play a critical role in its tumor-promoting potential. Recently we reported that a naturally occurring flavonoid, silibinin, protects UVB-induced skin damages and prevents photocarcinogenesis. Here we examined silibinin efficacy on acute and chronic UVB caused mitogen-activated protein kinases (MAPKs) and AKT activation and associated biological responses in SKH-1 hairless mouse skin. A single UVB exposure at 180 mJ/cm2 dose resulted in varying degrees of ERK1/2, JNK1/2, MAPK/p38 and AKT phosphorylation at various time-points in mouse skin; however, topical application of silibinin prior to or immediately after UVB exposure, or its dietary feeding strongly inhibited the activation of these molecules at all the time-points examined. Stronger effects of silibinin towards inhibition of UVB caused phosphorylation of MAPKs and AKT were also observed in a chronic UVB (180 mJ/cm2/day for 5 days) exposure protocol. Immunohistochemical analysis of chronically exposed skin sections showed that silibinin treatment in all three protocols increases UVB-induced p53-positive cells and decreases UVB-caused cell proliferation, apoptotic and sunburn cells. These findings suggest that silibinin inhibits UVB-induced MAPK and AKT signaling and increases p53 in mouse skin, and that these effects of silibinin possibly lead to a decrease in UVB-caused proliferation and apoptosis, which might, in part, be responsible for its overall efficacy against photocarcinogenesis. PMID- 15831528 TI - In vivo antitumor activity of the NF-kappaB inhibitor dehydroxymethylepoxyquinomicin in a mouse model of adult T-cell leukemia. AB - Adult T-cell leukemia (ATL) is an aggressive neoplasm caused by human T-cell leukemia virus type I (HTLV-I). The nuclear transcription factor, NF-kappaB, is induced by HTLV-I and is central to the ensuing neoplasia. To examine the effect of a novel NF-kappaB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), on ATL in vivo, we developed an improved severe combined immunodeficiency (SCID) mouse model for ATL. Five-week-old SCID mice in which natural killer (NK) cell activity had been eliminated were inoculated intraperitoneally with the HTLV-I-infected cell lines, TL-Om1, MT-1, MT-2 and HUT-102. No engraftment of TL-Om1 cells and little tumorigenesis of MT-1 cells were detected 40 days after injection. In contrast, inoculation of mice with MT-2 and HUT-102 cells elicited high mortality, 100% frequency of gross tumor formation and tumor cell infiltration of various organs, all of which were reduced by coadministration of DHMEQ during the inoculation. Moreover, tumors from mice treated with DHMEQ had a high frequency of apoptosis. These results suggest that DHMEQ induces apoptosis in HTLV-I transformed cells in vivo, resulting in inhibition of tumor formation and organ infiltration, thereby enhancing survival. PMID- 15831529 TI - The race associated allele of Semaphorin 3B (SEMA3B) T415I and its role in lung cancer in African-Americans and Latino-Americans. AB - SEMA3B has been implicated as important for neuronal development and as a tumor suppressor in lung cancer. A single nucleotide alteration of this gene leads to the amino acid substitution T415I, and functionally, this variant protein has a reduced ability to act as a tumor suppressor. The prevalence of this variant in populations is unclear and its role in inherited lung cancer susceptibility has not been tested. Utilizing case-control studies of head and neck squamous cell carcinoma in a Caucasian population and of lung cancer in African-American and Latino-American populations, we determined both the prevalence of this polymorphic variant and its association with the case status of these patients. The variant Ile allele occurs at an allele frequency of 0.18 in African-American and 0.39 in Latino-American control subjects but not in Caucasian subjects. In analyses controlling for ethnicity and known lung cancer risk factors, a significant association was observed between case status and possession of the variant allele (OR 0.71, 95% CI 0.51-0.99). In stratified analysis, both Latino Americans (OR 0.56, 95% CI 0.32-1.01) and African-Americans (OR 0.75, 95% CI 0.50 1.13) also showed a reduced risk of disease associated with the variant Ile allele. Possessing either the heterozygous or homozygous variant genotype confers a >40% reduced relative risk of lung cancer in Latino Americans controlling for other lung cancer risk factors. This study points to the need for further examination of this gene and its variant in lung cancer and other diseases. PMID- 15831530 TI - Modulation of aberrant crypt foci and apoptosis by dietary herbal supplements (quercetin, curcumin, silymarin, ginseng and rutin). AB - It is estimated that one-third of Americans use dietary herbal supplements on a regular basis. Diets rich in bioactive phytochemicals are associated with reduced risk of certain cancers, notably, colon cancer. Herbal supplements have not been directly tested as sources of bioactive cancer preventives. Hence, this study compares the ability of four herbal flavonoids (quercetin, curcumin, rutin and silymarin) and one whole herb mixture (ginseng powder) to suppress aberrant crypt foci (ACF) in an azoxymethane (AOM)-induced rat colon cancer model. Second, this study examines the effect of these herbal compounds on apoptosis and the mechanisms by which these compounds evoke apoptosis. The results of this study show that diets containing quercetin, curcumin, silymarin, ginseng and rutin decreased the number of ACFs by 4-, 2-, 1.8-, 1.5- and 1.2-fold, respectively compared with control. Histological analysis of the colon mucosa revealed that all the herbal supplements, except silymarin, induced apoptosis, with quercetin being the most potent (3x increase compared with control). Furthermore, ginseng and curcumin were region-specific in inducing apoptosis. The ability of quercetin and curcumin to modulate ACFs correlates well with their ability to induce apoptosis. Western blot analysis of caspase 9, Bax (proapoptotic) and Bcl-2 (antiapoptotic) proteins from the colon scraping suggests that quercetin and curcumin induce apoptosis via the mitochondrial pathway. Taken together, the results of this study suggest that these herbal supplements may exert significant and potentially beneficial effects on decreasing the amount of precancerous lesions and inducing apoptosis in the large intestine. PMID- 15831531 TI - Quantitative trait locus analysis reveals two intragenic sites that influence O6 alkylguanine-DNA alkyltransferase activity in peripheral blood mononuclear cells. AB - The repair of specific types of DNA alkylation damage by O6-alkylguanine-DNA alkyltransferase (MGMT) is a major mechanism of resistance to the carcinogenic and chemotherapeutic effects of certain alkylating agents. MGMT expression levels vary widely between individuals but the underlying causes of this variability are not known. To address this, we used an expressed single nucleotide polymorphism (SNP) and demonstrated that the MGMT alleles are frequently expressed at different levels in peripheral blood mononuclear cells (PBMC). This suggests that there is a genetic component of inter-allelic variation of MGMT levels that maps close to or within the MGMT locus. We then used quantitative trait locus (QTL) analysis using intragenic SNPs and found that there are at least two sites influencing inter-individual variation in PBMC MGMT activity. One is characterized by an SNP at the 3' end of the first intron and the second by two SNPs in the last exon. The latter are in perfect disequilibrium and both result in amino acid substitutions-one of them, Ile143Val, affecting an amino acid close to the Cys145 residue at the active site of MGMT. Using in vitro assays, we further showed that while the Val143 variant did not affect the activity of the protein on methylated DNA substrate, it was more resistant to inactivation by the MGMT pseudosubstrate, O6-(4-bromothenyl)guanine. These findings suggest that further investigations of the potential epidemiological and clinical significance of inherited differences in MGMT expression and activity are warranted. PMID- 15831532 TI - Non-nicotinic transmission in autonomic ganglia revisited--an important physiological function? PMID- 15831533 TI - Impaired voluntary running capacity of creatine kinase-deficient mice. AB - The creatine kinase system (CK) is important for energy delivery in skeletal and cardiac muscles. The two main isoforms of this enzyme, cytosolic MM-CK and mitochondrial mi-CK, are expressed in a developmental and muscle-type specific manner. Mice deficient in one or both of these isoforms are viable and fertile but exhibit profound functional, metabolic and structural muscle remodelling that primarily affects fast skeletal muscles, which show an increased contribution of oxidative metabolism to contractile function. However, the consequences of these alterations in terms of physical capabilities have not yet been characterized. Consequently, we compared the voluntary exercise capacity of 9-month-old male wild-type (WT), M-CK knockout (M-CK(-/-)), and M-CK and mi-CK double knockout (CK(-/-)) mice, using cages equipped with running wheels. Exercise performance, calculated by total distance covered and by work done during the training period, was more than 10-fold lower in CK(-/-) mice than controls, with M-CK(-/-) mice exhibiting intermediate performance. Similarly, the mean distance run per activation was lower in M-CK(-/-) and even lower in CK(-/-) mice. However, the maximal running speed (V(max)) was lower only for CK(-/-) mice. This was accompanied by severe skeletal muscle mass decrease in CK(-/-) mice, with signs of histological damage that included enlarged interstitial areas, aggregations of mononuclear cells in the interstitium, heterogeneity of myofibre size and the presence of very small fibres. No overt sign of cardiac dysfunction was observed by magnetic resonance imaging during dobutamine stimulation. These results show that metabolic failure induced by CK deficiency profoundly affects the ability of mice to engage in chronic bouts of endurance running exercise and that this decrease in performance is also associated with muscle wasting. PMID- 15831534 TI - Partitioning oxidative fuels during cold exposure in humans: muscle glycogen becomes dominant as shivering intensifies. AB - The effects of changes in shivering intensity on the relative contributions of plasma glucose, muscle glycogen, lipids and proteins to total heat production are unclear in humans. The goals of this study were: (1) to determine whether plasma glucose starts playing a more prominent role as shivering intensifies, (2) to quantify overall changes in fuel use in relation to the severity of cold exposure, and (3) to establish whether the fuel selection pattern of shivering is different from the classic fuel selection pattern of exercise. Using a combination of indirect calorimetry and stable isotope methodology, fuel metabolism was monitored in non-acclimatized adult men exposed for 90 mins to 10 degrees C (low-intensity shivering (L)) or 5 degrees C (moderate-intensity shivering (M)). Results show that plasma glucose oxidation is strongly stimulated by moderate shivering (+122% from L to M), but the relative contribution of this pathway to total heat generation always remains minor (< 15% of total heat production). Instead, muscle glycogen is responsible for most of the increase in heat production between L and M. By itself, the increase in CHO oxidation is responsible for the 100 W increase in metabolic rate observed between L and M, because rates of lipid and protein oxidation remain constant. This high reliance on CHO is not compatible with the well known fuel selection pattern of exercise, when considering the relatively low metabolic rates elicited by shivering (approximately 30% for M). We conclude that shivering and exercise of similar energy requirements appear to be supported by different fuel mixtures. Investigating the physiological mechanisms underlying why a muscle producing only heat (shivering), or significant movement (exercise), shows a different pattern of fuel selection at the same power output strikes us as a fascinating area for future research. PMID- 15831536 TI - Electrophysiological differences between nociceptive and non-nociceptive dorsal root ganglion neurones in the rat in vivo. AB - Intracellular recordings were made from 1022 somatic lumbar dorsal root ganglion (DRG) neurones in anaesthetized adult rats, classified from dorsal root conduction velocities (CVs) as C, Adelta or Aalpha/beta, and according to their responses to mechanical and thermal stimuli as nociceptive (including high threshold mechanoreceptive (HTM) units), and non-nociceptive (including low threshold mechanoreceptive (LTM) and cooling units). Of these, 463 met electrophysiological criteria for analysis of action potentials (APs) evoked by dorsal root stimulation. These included 47 C-, 71 Adelta- and 102 Aalpha/beta nociceptive, 10 C-, 8 Adelta- and 178 Aalpha/beta-LTM, 18 C- and 19 Adelta- unresponsive, and 4 C-cooling units. Medians of AP and afterhyperpolarization (AHP) durations and AP overshoots were significantly greater for nociceptive than LTM units in all CV groups. AP overshoots and AHP durations were similar in nociceptors of all CV groups whereas AP durations were greater in slowly conducting, especially C-fibre, nociceptors. C-cooling units had faster CVs, smaller AP overshoots and shorter AP durations than C-HTM units. A subgroup of Aalpha/beta-HTM, moderate pressure units, had faster CVs and AP kinetics than other Aalpha/beta-HTM units. Of the Aalpha/beta-LTM units, muscle spindle afferents had the fastest CV and AP kinetics, while rapidly adapting cutaneous units had the slowest AP kinetics. AP variables in unresponsive and nociceptive units were similar in both C- and Adelta-fibre CV groups. The ability of fibres to follow rapid stimulus trains (fibre maximum following frequency) was correlated with CV but not sensory modality. These findings indicate both the usefulness and limitations of using electrophysiological criteria for identifying neurones acutely in vitro as nociceptive. PMID- 15831535 TI - Transient outward potassium current, 'Ito', phenotypes in the mammalian left ventricle: underlying molecular, cellular and biophysical mechanisms. AB - At least two functionally distinct transient outward K(+) current (I(to)) phenotypes can exist across the free wall of the left ventricle (LV). Based upon their voltage-dependent kinetics of recovery from inactivation, these two phenotypes are designated 'I(to,fast)' (recovery time constants on the order of tens of milliseconds) and 'I(to,slow)' (recovery time constants on the order of thousands of milliseconds). Depending upon species, either I(to,fast), I(to,slow) or both current phenotypes may be expressed in the LV free wall. The expression gradients of these two I(to) phenotypes across the LV free wall are typically heterogeneous and, depending upon species, may consist of functional phenotypic gradients of both I(to,fast) and I(to,slow) and/or density gradients of either phenotype. We review the present evidence (molecular, biophysical, electrophysiological and pharmacological) for Kv4.2/4.3 alpha subunits underlying LV I(to,fast) and Kv1.4 alpha subunits underlying LV I(to,slow) and speculate upon the potential roles of each of these currents in determining frequency dependent action potential characteristics of LV subepicardial versus subendocardial myocytes in different species. We also review the possible functional implications of (i) ancillary subunits that regulate Kv1.4 and Kv4.2/4.3 (Kvbeta subunits, DPPs), (ii) KChIP2 isoforms, (iii) spider toxin mediated block of Kv4.2/4.3 (Heteropoda toxins, phrixotoxins), and (iv) potential mechanisms of modulation of I(to,fast) and I(to,slow) by cellular redox state, [Ca(2)(+)](i) and kinase-mediated phosphorylation. I(to) phenotypic activation and state-dependent gating models and molecular structure-function relationships are also discussed. PMID- 15831537 TI - Differential effects of maurocalcine on Ca2+ release events and depolarization induced Ca2+ release in rat skeletal muscle. AB - Maurocalcine (MCa), a 33 amino acid toxin obtained from scorpion venom, has been shown to interact with the isolated skeletal-type ryanodine receptor (RyR1) and to strongly modify its calcium channel gating. In this study, we explored the effects of MCa on RyR1 in situ to establish whether the functional interaction of RyR1 with the voltage-sensing dihydropyridine receptor (DHPR) would modify the ability of MCa to interact with RyR1. In developing skeletal muscle cells the addition of MCa into the external medium induced a calcium transient resulting from RyR1 activation and strongly inhibited the effect of the RyR1 agonist chloro m-cresol. In contrast, MCa failed to affect the depolarization-induced Ca(2+) release. In intact adult fibres MCa did not induce any change in the cytosolic Ca(2+) concentration. However, when the surface membrane was permeabilized and calcium release events were readily observable, MCa had a time-dependent dual effect: it first increased event frequency, from 0.060 +/- 0.002 to 0.150 +/- 0.007 sarcomere(-1) s(-1), and reduced the amplitude of individual events without modifying their spatial distribution. Later on it induced the appearance of long lasting events resembling the embers observed in control conditions but having a substantially longer duration. We propose that the functional coupling of DHPRs and RyR1s within a Ca(2+) release unit prevents MCa from either reaching its binding site or from being able to modify the gating not only of the RyR1s physically coupled to DHPRs but all RyR1s within the Ca(2+) release unit. PMID- 15831538 TI - Rapid protein kinase C-dependent reduction of rat skeletal muscle voltage-gated sodium channels by ciliary neurotrophic factor. AB - The ciliary neurotrophic factor (CNTF), known to exert long-term myotrophic effects, has not yet been shown to induce a rapid biological response in skeletal muscles. The present in vitro study gives rise to the possibility that CNTF could affect the sodium channel activity implied in the triggering of muscle fibre contraction. Therefore, we investigated the effects of an external CNTF application on macroscopic sodium current (I(Na)) in rat native fast-twitch skeletal muscle (flexor digitorum brevis, FDB) by using a cell-attached patch clamp technique. The I(Na) peak amplitude measured at a depolarizing pulse from 100 to -10 mV is rapidly reduced in a time- and dose-dependent manner by CNTF (0.01-20 ng ml(-1)). The maximal decrease is 25% after 10 min incubation in 2 ng ml(-1) CNTF. There was no alteration in activation or inactivation kinetics, or in activation curves constructed from current-voltage relationships in the presence of CNTF. In contrast, the relative I(Na) inhibition induced by CNTF is accompanied by a hyperpolarizing shift in the midpoint of the inactivation curves: -6 and -10 mV for the steady-state fast and slow inactivation, respectively. Furthermore, CNTF induces a 5 mV hyperpolarization of the resting membrane potential of the fibres. The effects of CNTF are similar to those of 1 oleoyl-2-acetyl-sn-glycerol (OAG), a protein kinase C (PKC) activator, when no effect is observed in the presence of chelerythrine, a PKC inhibitor. These results suggest that, in skeletal muscle, CNTF can rapidly decrease sodium currents by altering inactivation gating, probably through an intracellular PKC dependent mechanism that could lead to decreased membrane excitability. The present study contributes to a better understanding of the physiological role of endogenous CNTF. PMID- 15831539 TI - In vivo and in vitro functional characterization of Andersen's syndrome mutations. AB - The inward rectifier K(+) channel Kir2.1 carries all Andersen's syndrome mutations identified to date. Patients exhibit symptoms of periodic paralysis, cardiac dysrhythmia and multiple dysmorphic features. Here, we report the clinical manifestations found in three families with Andersen's syndrome. Molecular genetics analysis identified two novel missense mutations in the KCNJ2 gene leading to amino acid changes C154F and T309I of the Kir2.1 open reading frame. Patch clamp experiments showed that the two mutations produced a loss of channel function. When co-expressed with Kir2.1 wild-type (WT) channels, both mutations exerted a dominant-negative effect leading to a loss of the inward rectifying K(+) current. Confocal microscopy imaging in HEK293 cells is consistent with a co-assembly of the EGFP-fused mutant proteins with WT channels and proper traffick to the plasma membrane to produce silent channels alone or as hetero-tetramers with WT. Functional expression in C2C12 muscle cell line of newly as well as previously reported Andersen's syndrome mutations confirmed that these mutations act through a dominant-negative effect by altering channel gating or trafficking. Finally, in vivo electromyographic evaluation showed a decrease in muscle excitability in Andersen's syndrome patients. We hypothesize that Andersen's syndrome-associated mutations and hypokalaemic periodic paralysis associated calcium channel mutations may lead to muscle membrane hypoexcitability via a common mechanism. PMID- 15831540 TI - Acute dopamine/noradrenaline reuptake inhibition enhances human exercise performance in warm, but not temperate conditions. AB - Nine healthy endurance-trained males were recruited to examine the effect of a dual dopamine/noradrenaline reuptake inhibitor on performance, thermoregulation and the hormonal responses to exercise. Subjects performed four trials, ingesting either a placebo (pla) or 2 x 300 mg bupropion (bup), prior to exercise in temperate (18 degrees C) or warm (30 degrees C) conditions. Trials consisted of 60 min cycle exercise at 55% W(max) immediately followed by a time trial (TT). TT performance in the heat was significantly improved by bupropion (pla: 39.8 +/- 3.9 min, bup: 36.4 +/- 5.7 min; P = 0.046), but no difference between treatments was apparent in temperate conditions (pla: 30.6 +/- 2.2 min, bup: 30.6 +/- 1.9 min; P = 0.954). While TT power output was consistently lower in the heat when compared to temperate conditions, this decrement was attenuated by bupropion. At the end of the TT in the heat, both core temperature (pla 39.7 +/- 0.3 degrees C, bup 40.0 +/- 0.3 degrees C; P = 0.017) and HR (pla 178 +/- 7 beats min(-1), bup 183 +/- 12 beats min(-1); P = 0.039), were higher in the bupropion trial than in the placebo. Circulating pituitary and adrenal hormone concentrations increased throughout exercise in all trials. Circulating serum prolactin was elevated above temperate levels during exercise in a warm environment (P < 0.001). These data indicate that performance in warm conditions is enhanced by acute administration of a dual dopamine/noradrenaline reuptake inhibitor. No such effect was apparent under temperate conditions. It appears that bupropion enabled subjects to maintain a greater TT power output in the heat with the same perception of effort and thermal stress reported during the placebo trial, despite the attainment of a higher core temperature. PMID- 15831541 TI - Guidelines for preoperative assessment: impact on clinical practice and costs. AB - OBJECTIVE: To describe preoperative evaluation in the San Giovanni Battista Hospital in Turin and to forecast the economic impact when preoperative assessment guidelines are implemented. DESIGN: We enrolled, in a month, 702 consecutive patients, excluding cardiac, thoracic, neuro- and vascular surgery, as well as emergency operations. Preoperative assessment data were collected individually, followed by simulating various applications of guidelines based on: (i) preoperative tests relying on full medical history and physical examination to discriminate preoperative risk patients; (ii) organization of a preoperative evaluation unit and tests before patient hospitalization. MAIN MEASURES: Mean number of tests prescribed, preoperative assessment cost per patient. RESULTS: The application of preoperative guidelines would decrease the mean number of tests prescribed from 20 laboratory and 1.9 instrumental to, respectively, 3 and 1.4 per patient. Tests deemed inappropriate by guidelines did not add any relevant clinical information to our study. Economic analysis estimates a reduction of 63% in cost per patient for preoperative tests by introducing guideline criteria (from 69 euros to 26 euros). As regards the cost per patient for preoperative evaluation and hospital stay (115 euros considering only variable costs, 580 euros including all costs), the application of the guidelines would reduce costs by 41-52% according to different cost evaluation approaches for hospital stay. CONCLUSION: Preoperative guidelines fully introduced in practice could notably increase efficiency without affecting the quality of care. PMID- 15831542 TI - Failure of Internet-based audit and feedback to improve quality of care delivered by primary care residents. AB - OBJECTIVE: To determine the effectiveness of Internet-based audit and feedback to physicians to improve care for diabetes and hypertension. DESIGN: Time-series analysis of an intervention. METHODS: The study setting was Harvard Vanguard Medical Associates, a 14-site multispecialty group in greater Boston. The study period was July 1997-June 1999. PARTICIPANTS: were 12 primary care internal medicine residents who provided care to adult patients with diabetes (n = 76 pre intervention and n = 88 post-intervention), hypertension (n = 329 pre intervention and n = 338 post-intervention), or both (n = 62 pre-intervention and n = 71 post-intervention). We determined the proportion of each resident's patients whose care fulfilled national guidelines for quality (i.e. diabetes patients had hemoglobin testing in the previous 6 months or hypertension patients received a beta-blocker or diuretic in the same time period). After meeting individually with each resident to obtain informed consent and to encourage participation, we sent each resident information for accessing his or her practice profile on a secure website. The main outcome measures were (i) the proportion of resident physicians who accessed their profiles and (ii) change following the intervention in the proportion of patients whose care followed national guidelines. RESULTS: Over a 1-year period, only four of the 12 residents accessed their websites. One of the residents visited her site three times, while the other three residents visited their sites once each. In interrupted time series analyses, the intervention had no discernible effect on adherence to practice guidelines for diabetes or hypertension. CONCLUSION: The lack of participation in this Internet-based intervention may have important implications for the development of future programs that require physicians to interact with technology to improve quality of care. PMID- 15831543 TI - A controlled before-after trial of structured diabetes care in primary health centres in a newly developed country. AB - OBJECTIVE: To evaluate the long-term impact of a structured approach to improving the quality of diabetes care in general practice in the United Arab Emirates. DESIGN: Controlled before-after trial within a health district with three primary health centres (PHCs) in the intervention group and the six remaining serving as controls. Outcomes and adherence to guidelines were measured over the year before the intervention began and for a second 1-year period at the end of the intervention period. Data were collected by chart abstraction. SETTING: The study was performed in PHCs in the United Arab Emirates, a newly developed country on the Arabian peninsula. STUDY PARTICIPANTS: Subjects continuously followed in nine PHCs for diabetes care for the period of the study (N = 738) were included in the study. INTERVENTION: Structured diabetes care, including the development of general practice diabetes clinics, a patient education program, a health care professional education program, and improved recording of clinical information, was provided for the 33-month time period. RESULTS: There was a statistically significant improvement in three of the process of care variables (ordering HbA1c, cholesterol, and documenting foot examinations) whereas the four remaining variables did not improve. There was limited impact on outcome variables. CONCLUSIONS: The intervention described in this study demonstrated an improvement in some process of care measures suggesting an impact of this type of delivery model in this environment. PMID- 15831544 TI - Quality improvement programme on the frontline: an International Committee of the Red Cross experience in the Democratic Republic of Congo. AB - BACKGROUND: Majority of research in Quality Improvement, focuses on developed countries or development programs. Humanitarian organisations frequently work in developing countries, often in emergency situations with rapid staff turnover. Objectives of this study are twofold: first to develop a methodology of motivation and restoration of collapsed health structures through the creation of community based QI indicators; second, to implement these indicators to improve quality of care. METHODOLOGY: Using a community-based approach, the International Committee of the Red Cross (ICRC) team together with local health committee and nurses developed quality indicators divided into six different categories. Of 16 community primary health centers and four hospitals supported by ICRC, six health centers and one hospital were chosen to follow quality indicators for three of six indicator categories. Initial data were collected in January 2003 and compared with data serially gathered throughout the year. RESULTS: In the category rational prescription, all health facilities except for one showed improvement in every category. In the hygiene category, four of seven health structures showed 100% improvement in their score. Three of seven facilities showed impressive improvement in the category pharmacy management. CONCLUSION: Involving the community to design population based indicators helped communities take ownership of the indicators. Our findings that poor performance on indicators prompted communities to seek training and assistance to improve quality of care emphasized this. Continued adherence and improvement in each category confirmed the long term effects of teaching sessions in the areas of rational prescription, hygiene and pharmacy maintenance. PMID- 15831545 TI - The DNA damage response and patient safety: engaging our molecular biology oriented colleagues. AB - The imperative to improve patient safety is clear. Biomedical scientists, who account for a large proportion of medical school faculty, and clinicians tend to speak different languages. Biological systems are remarkable for their high robustness, flexibility, and efficiency. Biomedical scientists possess a profound understanding of the complex mechanisms that govern organisms. Their insights may inform the design of safer health care systems. We propose a model to assist in bi-directional communication between these disciplines. We use the principles and mechanisms of the DNA damage response to describe the central concepts of safety science and discuss similarities and differences between the systems of DNA repair and organizational approaches to safety in health care. We suggest that such biomedical scientists can and should be engaged in the effort to bring education about patient safety management into the medical school curriculum and to make patient care safer. PMID- 15831546 TI - Measuring the quality of hospital tuberculosis services: a prospective study in four Zimbabwe hospitals. AB - OBJECTIVE: To show how the use of a prospective approach to measuring the quality of services for a specific diagnosis can generate useful information for improving the quality of services in environments with limited information technology and data. DESIGN: Tracer approach focusing on intensive treatment of tuberculosis in hospital. The study was conducted in Zimbabwe in 1999. Local tuberculosis management guidelines were first translated into explicit quality assessment criteria and a panel of public health experts assisted in weighting different factors (structural and process) of the criteria. Factor weightings were based on both local knowledge and experience, and potential contribution of a factor to the likelihood of a positive outcome. A total of 138 patients was recruited into the study cohort at admission and followed up to discharge. An assessment of what was done to and for the patient was made for the entire hospitalization episode using explicit criteria. Comparisons were made between actual and maximum performance scores. SETTING: The study was conducted at four regional referral hospitals. The hospitals serve at least six secondary hospitals, and several public and private primary care facilities. The hospitals have a dual role as they also provide secondary care to their immediate catchment population. RESULTS: Notable quality gaps are observed between actual and maximum quality levels in all four hospitals although the size of the gap differed significantly. Variation in the quality of services between the hospitals is explained by distinguishable differences in structural and process aspects of tuberculosis management. CONCLUSIONS: It is feasible to conduct prospective quality assessment in developing countries with minimal disruption of routine activities. The study also showed that prospective exploration of health care quality for a specific diagnosis can provide insights into hospital-level quality issues. Such information is useful for monitoring and improving the quality of hospital services in general. PMID- 15831547 TI - Using concept mapping to design an indicator framework for addiction treatment centres. AB - OBJECTIVE: The objective of this study is to determine an indicator framework for addiction treatment centres based on the demands of stakeholders and in alignment with the European Foundation for Quality Management (EFQM) Excellence Model. SETTING: The setting is the Jellinek Centre based in Amsterdam, the Netherlands, which serves as a prototype for an addiction treatment centre. METHOD: Concept mapping was used in the construction of the indicator framework. During the 1-day workshop, 16 stakeholders generated, prioritized and sorted 73 items concerning quality and performance. Multidimensional scaling and cluster analysis was applied in constructing a framework consisting of two dimensions and eight clusters. RESULTS: The horizontal axis of the indicator framework is named 'Organization' and has two poles, namely, 'Processes' and 'Results'. The vertical axis is named ' Task' and the poles are named 'Efficient treatment' and 'Prevention programs'. The eight clusters in the two-dimensional framework are arranged in the following, prioritized sequence: 'Efficient treatment network', 'Effective service', ' Target group', 'Quality of life', 'Efficient service', 'Knowledge transfer', 'Reducing addiction related problems', and 'Prevention programs'. The most important items in the framework are: 'patients are satisfied with their treatment', 'early interventions', and 'efficient treatment chain'. CONCLUSION: The indicator framework aligns with three clusters of the results criteria of the EFQM Excellence Model. It is based on the stakeholders' perspectives and is believed to be specific for addiction treatment centres. The study demonstrates that concept mapping is a suitable strategy for generating indicator frameworks. PMID- 15831548 TI - Results of a national asthma campaign survey of primary care in Scotland. AB - OBJECTIVE: To identify within primary care in Scotland how far procedures for asthma review and patient education match guideline recommendations. DESIGN AND SETTING: Telephone survey of a one in four stratified random sample of all 1058 general practices in Scotland. PARTICIPANTS: Practice nurses, general practitioners. MAIN OUTCOME MEASURES: Number of practices matching guideline recommendations for asthma review, targeting of care, use of structured asthma records, provision of management plans, education, and regular audit. RESULTS: Of 276 general practices contacted 91% (251) completed the questionnaire; 93% (228) ran an asthma review service; 74% (166) employed a specially trained asthma nurse; 39% (106) had a policy for providing action plans; 63% (155) had carried out an asthma audit in the previous 3 years; 76% (218) used a structured tool in consultations, 46% with use of computer technology, 34% used only a manual stamp. Sixty-six per cent (173) had searched for patients overusing beta2 agonists; 32% (79) had searched for patients on medication treatment step 3 and above. Single- or two-partner practices were less likely to follow guideline recommendations but neither rurality nor deprivation was related to guideline compliance. CONCLUSIONS: Three-quarters of Scottish general practices have trained asthma nurses and offer patients asthma review, but only a minority have proactive care procedures for targeting patients or a policy for providing patients with action plans. Practice systems are underused for identifying 'at-risk' patients. There is a need for proactive procedures and provision of self-management materials to patients. Access to trained asthma nurses needs to be improved. PMID- 15831549 TI - Central venous stenosis as a complication of ipsilateral haemodialysis fistula and pacemaker. PMID- 15831550 TI - Thyroid function and glomerular filtration--a potential for Grave errors. PMID- 15831551 TI - A glowing report on dialysis. PMID- 15831552 TI - Bilateral calcified renal artery aneurysms in a patient with von Recklinghausen's disease. PMID- 15831553 TI - Scattered hyperdense lesions in a haemodialysis patient. PMID- 15831554 TI - Postpartum renal cortical necrosis. PMID- 15831555 TI - High plasma N-terminal pro-brain natriuretic peptide level found in diabetic patients after myocardial infarction is associated with an increased risk of in hospital mortality and cardiogenic shock. AB - AIMS: No studies have yet been conducted concerning plasma N-terminal pro-brain natriuretic peptide (Nt-pro-BNP) levels after Myocardial Infarction (MI) and their relationship with short-term outcomes in diabetic patients. METHODS AND RESULTS: Five hundred and sixty patients hospitalized for MI from the RICO survey, including 199 diabetic and 361 non-diabetic subjects, were included in the study. Plasma Nt-pro-BNP levels were measured on admission. Median Nt-pro-BNP levels were significantly higher in diabetic patients compared with non-diabetic patients [245 (81-77) vs. 130 (49-199) pmol/L, P<0.0001]. This difference remained highly significant after adjustment for age, female gender, creatinine clearance, left ventricular ejection fraction (LVEF), plasma peak troponin, anterior wall necrosis, and hypertension. In multivariable analysis, Nt-pro-BNP levels were negatively associated with creatinine clearance (P<0.0001) and LVEF (P<0.0001) and positively associated with plasma peak troponin (P<0.0001), age (P=0.0029), diabetes (P=0.0031), and female gender (P=0.0102). Diabetic patients showed a 4.7-fold increase in hospital mortality (15.6 vs. 3.3%, P<0.0001) and a 2.2-fold increase in cardiogenic shock (17.6 vs. 7.7%, P=0.0004). In multivariable analysis, diabetes was an independent factor for mortality [OR: 1.79 (1.45-2.20); P=0.0064] and cardiogenic shock [OR: 1.45 (1.22-1.72); P=0.0364] when the variable Nt-pro-BNP level was not introduced into the model, but was less significantly associated with mortality [OR: 1.73 (1.39-2.16); P=0.0107] and no longer associated with cardiogenic shock when Nt-pro-BNP was in the model. CONCLUSION: After MI, diabetes is independently associated with high plasma Nt-pro-BNP levels. This elevated Nt-pro-BNP is strongly associated with the increased incidence of in-hospital mortality and cardiogenic shock observed in diabetes. Our findings clearly indicate that plasma Nt-pro-BNP provides highly valuable prognostic information on in-hospital outcome after MI, in particular in diabetic patients. PMID- 15831556 TI - The reproducibility and sensitivity of the 6-min walk test in elderly patients with chronic heart failure. AB - AIMS: The 6-min walk test (6-MWT) is used to estimate functional capacity. However, in elderly patients with chronic heart failure (CHF): (i) 1 year reproducibility of the 6-MWT; (ii) sensitivity of the 6-MWT to self-perceived changes in symptoms of heart failure; and (iii) implications for patient numbers required for studies using the 6-MWT as an endpoint have not been described. METHODS AND RESULTS: One thousand and seventy-seven patients with CHF, aged>60, with NYHA Class > or =II were recruited. Heart failure symptom assessment was determined using a questionnaire related to aspects of physical function, and patients performed a baseline 6-MWT, with follow-up 1 year later. Seventy-four patients with unchanged symptoms had an unchanged 6-MWT distance, with an overall intraclass correlation coefficient of 0.80 (95% CI=0.69-0.87). Four hundred and twenty-three patients reported an improvement in symptoms during follow-up. There was a negative correlation (r=-0.55; P=0.0001) between Delta symptoms and Delta 6 MWT (i.e. a reduced 6-MWT distance is associated with reduced symptom severity at follow-up). Five hundred and sixteen patients reported worsening symptoms of heart failure, a moderate inverse correlation (r=-0.53; P=0.0001) was displayed between Delta symptoms and Delta 6-MWT. For all patients, irrespective of symptom status, a high inverse correlation (r=-0.75; P=0.0001) was evident. On the basis of the data for patients with unchanged symptoms, it is calculated that to detect an increase in 6-MWT of 50 m, with 90% power, a study size of approximately 120 is required. CONCLUSION: In elderly patients with CHF, the 6-MWT shows satisfactory agreement when repeated 1 year later. Change in 6-MWT distance is sensitive to change in self-perceived symptoms of heart failure. PMID- 15831557 TI - Delayed enhancement cardiovascular magnetic resonance assessment of non-ischaemic cardiomyopathies. AB - Non-ischaemic cardiomyopathies (NICMs) are chronic, progressive myocardial diseases with distinct patterns of morphological, functional, and electrophysiological changes. In the setting of cardiomyopathy (CM), determining the exact aetiology is important because the aetiology is directly related to treatment and patient survival. Determining the exact aetiology, however, can be difficult using currently available imaging techniques, such as echocardiography, radionuclide imaging or X-ray coronary angiography, since overlap of features between CMs may be encountered. Cardiovascular magnetic resonance (CMR) imaging has recently emerged as a new non-invasive imaging modality capable of providing high-resolution images of the heart in any desired plane. Delayed contrast enhanced CMR (DE-CMR) can be used for non-invasive tissue characterization and may hold promise in differentiating ischaemic from NICMs, as the typical pattern of hyperenhancement can be classified as 'ischaemic-type' or 'non-ischaemic type' on the basis of pathophysiology of ischaemia. This article reviews the potential of DE-CMR to distinguish between ischaemic and NICM as well as to differentiate non-ischaemic aetiologies. Rather than simply describing various hyperenhancement patterns that may occur in different disease states, our goal will be (i) to provide an overall imaging approach for the diagnosis of CM and (ii) to demonstrate how this approach is based on the underlying relationships between contrast enhancement and myocardial pathophysiology. PMID- 15831558 TI - Regulation of matrix metalloproteinase-9 release from IL-8-stimulated human neutrophils. AB - Matrix metalloproteinase-9 (MMP-9) is present in the tertiary granules of neutrophils and can be released following stimulation. We examined the signaling mechanisms that regulate interleukin-8 (IL-8)-mediated MMP-9 release from neutrophils. IL-8 activates neutrophils by interacting with two receptors: CXC chemokine receptor 1 (CXCR1) and CXCR2. Blocking CXCR1 had no effect on IL-8 mediated MMP-9 release, whereas blocking CXCR2 significantly reduced MMP-9 release. We also found that stimulating CXCR2 alone was sufficient to induce MMP 9 release. This process was independent of changes in the intracellular calcium concentration. Src-family kinases and protein kinase C (PKC) were involved in two mutually exclusive pathways regulating IL-8-mediated MMP-9 release. Inhibition of extracellular signal-regulated kinase (ERK)1/2 blocked IL-8-mediated MMP-9 release; however, inhibition of p38 mitogen-activated protein kinase had no effect on MMP-9 release. We found ERK1/2 was activated downstream of PKC, but not Src-family kinases, in this system. These data suggest that IL-8-induced MMP-9 release from neutrophils is mediated through CXCR2 and involves two distinct pathways, one involving PKC and ERK1/2 and the other involving Src-family kinases. Furthermore, our data show that the mechanisms that regulate MMP-9 release from tertiary granules are different from those that regulate primary granule release. PMID- 15831559 TI - MIP-1alpha[CCL3] acting on the CCR1 receptor mediates neutrophil migration in immune inflammation via sequential release of TNF-alpha and LTB4. AB - In the present study, we investigated the involvement of macrophage-inflammatory protein-1alpha (MIP-1alpha)[CC chemokine ligand 3 (CCL3)], MIP-1beta[CCL4], regulated on activation, normal T expressed and secreted (RANTES)[CCL5], and CC chemokine receptors (CCRs) on neutrophil migration in murine immune inflammation. Previously, we showed that ovalbumin (OVA)-triggered neutrophil migration in immunized mice depends on the sequential release of tumor necrosis factor alpha (TNF-alpha) and leukotriene B(4)(LTB(4)). Herein, we show increased mRNA expression for MIP-1alpha[CCL3], MIP-1beta[CCL4], RANTES[CCL5], and CCR1 in peritoneal cells harvested from OVA-challenged, immunized mice, as well as MIP 1alpha[CCL3] and RANTES[CCL5] but not MIP-1beta[CCL4] proteins in the peritoneal exudates. OVA-induced neutrophil migration response was muted in immunized MIP 1alpha[CCL3](-/-) mice, but it was not inhibited by treatment with antibodies against RANTES[CCL5] or MIP-1beta[CCL4]. MIP-1alpha[CCL3] mediated neutrophil migration in immunized mice through induction of TNF-alpha and LTB(4) synthesis, as these mediators were detected in the exudates harvested from OVA-challenged immunized wild-type but not MIP-1alpha[CCL3](-/-) mice; administration of MIP 1alpha[CCL3] induced a dose-dependent neutrophil migration, which was inhibited by treatment with an anti-TNF-alpha antibody in TNF receptor 1 (p55(-/-)) deficient mice or by MK 886 (a 5-lipoxygenase inhibitor); and MIP-1alpha[CCL3] failed to induce LTB(4) production in p55(-/-) mice. MIP-1alpha[CCL3] used CCR1 to promote neutrophil recruitment, as OVA or MIP-1alpha[CCL3] failed to induce neutrophil migration in CCR1(-/-) mice, in contrast to CCR5(-/-) mice. In summary, we have demonstrated that neutrophil migration observed in this model of immune inflammation is mediated by MIP-1alpha[CCL3], which via CCR1, induces the sequential release of TNF-alpha and LTB(4). Therefore, whether a similar pathway mediates neutrophil migration in human immune-inflammatory diseases, the development of specific CCR1 antagonists might have a therapeutic potential. PMID- 15831560 TI - Characterization of CCL20 secretion by human epithelial vaginal cells: involvement in Langerhans cell precursor attraction. AB - Mucosa represents the main site of pathogen/cell interactions. The two main types of cells forming the epithelial structure [epithelial cells and Langerhans cells (LC)] coordinate the first defense responses to avoid infection. To evaluate the involvement of epithelial cells in the early steps leading to a specific adaptive immune response, we have studied the interactions between vaginal epithelial and LC through the establishment of a human vaginal epithelial mucosa. We demonstrate that normal human vaginal epithelial cells constitutively secrete the chemokine macrophage inflammatory protein 3alpha/CC chemokine ligand 20 (CCL20), known to recruit LC precursors (LCps) selectively via its cognate CC chemokine receptor 6 (CCR6). This secretion is up-regulated by the proinflammatory cytokine interleukin-1beta through the nuclear factor-kappaB pathway. Similar results were obtained with the human vaginal epithelial cell line SiHa, which displays numerous homologies with normal vaginal cells. The chemotactic activity of the secreted CCL20 was demonstrated by its ability to attract LCp CCR6+. Moreover, the use of neutralizing polyclonal antibodies directed against the CCL20 molecule abolished this migration completely, suggesting that CCL20 is the main attracting factor for LCps, which is produced by the vaginal cells. These data indicate that vaginal epithelial cells play an important role in the immunological defense by attracting immune cells to the site of epithelial/pathogen contact. PMID- 15831561 TI - Covariance components models for longitudinal family data. AB - A longitudinal family study is an epidemiological design that involves repeated measurements over time in a sample that includes families. Such studies, that may also include relative pairs and unrelated individuals, allow closer investigation of not only the factors that cause a disease to arise, but also the genetic and environmental determinants that modulate the subsequent progression of that disease. Knowledge of such determinants may pay high dividends in terms of prognostic assessment and in the development of new treatments that may be tailored to the prognostic profile of individual patients. Unfortunately longitudinal family studies are difficult to analyse. They conflate the complex within-family correlation structure of a cross-sectional family study with the correlation over time that is intrinsic to longitudinal repeated measures. Here we describe an approach to analysis that is relatively straightforward to implement, yet is flexible in its application. It represents a natural extension of a Gibbs-sampling-based approach to the analysis of cross-sectional family studies that we have described previously. The approach can be applied to pedigrees of arbitrary complexity. It is applicable to continuous traits, repeated binary disease states, and repeated counts or rates with a Poisson distribution. It not only supports the analysis of observed determinants, including measured genotypes, but also allows decomposition of the correlation structure, thereby permitting conclusions to be drawn about the effect of unobserved genes and environment on key features of disease progression, and hence to estimate the heritability of these features. We demonstrate the efficacy of our methods using a range of simulated data analyses, and illustrate its practical application to longitudinal blood pressure data measured in families from the Framingham Heart Study. PMID- 15831563 TI - Commentary: Height and intelligence. PMID- 15831564 TI - Self-reported economic difficulties and coronary events in men: evidence from the Whitehall II study. AB - BACKGROUND: Numerous studies have demonstrated social inequalities in coronary heart disease using a variety of measures of social position. In this study we examine associations between persistent economic difficulties and serious coronary events. Our aim is to assess whether these associations are (i) explained by other measures of socioeconomic status, and (ii) mediated by psychosocial, behavioural and biological factors. METHODS: The data come from 5021 middle-aged, white-collar men in the Whitehall II study. Self-reported household financial problems, measured at baseline (1985-88) and Phase 3 (1991 93), were used to construct a five-category score of persistent economic difficulties. Associations between economic difficulties and incident coronary events were determined over an average follow-up of 7 years. Other socioeconomic, psychosocial, behavioural and biological explanatory variables were obtained from the Phase 3 questionnaire and clinical examination. RESULTS: Age-adjusted Cox regression analyses demonstrated steep gradients in the incidence of coronary events with economic difficulties. The relative hazard between the bottom and the top of the difficulties hierarchy was 2.5 (95% confidence intervals (CI) 1.2-5.2) for fatal and non-fatal myocardial infarction (MI), 2.1 (1.3-3.6) for MI plus definite angina and 2.8 (1.9-4.2) for total coronary events. Adjustment for other markers of socioeconomic position, early life factors, psychosocial work environment characteristics and health-related behaviours had little effect, while adjustment for the biological factors reduced the association between difficulties and coronary events by 16-24%. CONCLUSION: We have demonstrated an economic difficulties gradient in coronary events in men that is independent of other markers of socioeconomic position and appears to be only partially mediated by well-known risk factors in mid-life. PMID- 15831565 TI - The effects of socioeconomic development on worldwide hepatitis A virus seroprevalence patterns. AB - BACKGROUND: Hepatitis A virus (HAV) infection confers long-term immunity, so mathematical analysis of age-specific seroprevalence in populations can reveal changes in the infection rate over time. HAV transmission is related to access to clean drinking water, personal hygiene and public sanitation. METHODS: We used an SIR (susceptible-infectious-recovered) compartmental model with age structure to fit a time-dependent logistic function for HAV force of infection for 157 published age-seroprevalence data sets. We then fit linear regression models for socioeconomic variables and infection rate. RESULTS: The proportion of the population with access to clean drinking water, the value of the human development index (HDI), and per capita gross domestic product (GDP) are all inverse predictors of HAV infection rates. Declining infection rates were observed in 65.6% of the surveys. Discussion This work demonstrates the utility of HAV seroprevalence studies to reveal patterns of change in force of infection and to assess the association between socioeconomic risk factors and transmission rates. PMID- 15831566 TI - A comparison of genetic and environmental variance structures for asthma, hay fever and eczema with symptoms of the same diseases: a study of Norwegian twins. AB - BACKGROUND: We compared patterns of genetic and environmental influences on variation in liability for asthma, hay fever and eczema with those for symptoms of the same diseases, and determined how common sets of genes and environmental factors contribute to the relationship between diseases and symptoms among Norwegian twins. METHODS: Analyses were based on self-reported asthma, hay fever and eczema and symptoms of the same diseases among 3334 pairs of Norwegian twins aged 18-35 years. Structural equation modelling was conducted to estimate the genetic and environmental variance structures. RESULTS: For all diseases the concordances and the twin correlations were higher among monozygotic than among dizygotic twins. The results of the modelling confirmed that genetic effects were substantial for the diseases, and were more moderate for symptoms. The phenotypic correlation between disease and symptom was 0.67 for asthma and wheeze (a/w), 0.64 for hay fever and sneeze (hf/s), and 0.54 for eczema and itch (e/i). Decomposition of these correlations into genetic (G) and environmental (E) pathways revealed that G = 0.48 and E = 0.19 for a/w, G = 0.40 and E = 0.24 for hf/s, and G = 0.34 and E = 0.20 for e/i. For the diseases, the specific sources of genetic variance accounted for more variation than the specific environmental variance. Variance decomposition revealed that specific sources of variance were primarily explained by genetic effects for diseases and by environmental influences for symptoms. CONCLUSIONS: Genetic effects account for greater variation in reported diseases than symptoms. Co-occurrence of diseases and symptoms is mainly explained by genetic effects common to both phenotypes, but non-shared environment is also important. PMID- 15831567 TI - Differential regulation of KiSS-1 mRNA expression by sex steroids in the brain of the male mouse. AB - Kisspeptins are products of the Kiss1 gene, which bind to GPR54, a G protein coupled receptor. Kisspeptins and GPR54 have been implicated in the neuroendocrine regulation of GnRH secretion. To test the hypothesis that testosterone regulates Kiss1 gene expression, we compared the expression of KiSS 1 mRNA among groups of intact, castrated, and castrated/testosterone (T)-treated male mice. In the arcuate nucleus (Arc), castration resulted in a significant increase in KiSS-1 mRNA, which was completely reversed with T replacement, whereas in the anteroventral periventricular nucleus, the results were the opposite, i.e. castration decreased and T increased KiSS-1 mRNA expression. In the Arc, the effects of T on KiSS-1 mRNA were completely mimicked by estrogen but only partially mimicked by dihydrotestosterone, a nonaromatizable androgen, suggesting that both estrogen receptor (ER) and androgen receptor (AR) play a role in T-mediated regulation of KiSS-1. Studies of the effects of T on KiSS-1 expression in mice with either a deletion of the ERalpha or a hypomorphic allele to the AR revealed that the effects of T are mediated by both ERalpha and AR pathways, which was confirmed by the presence of either ERalpha or AR coexpression in most KiSS-1 neurons in the Arc. These observations suggest that KiSS-1 neurons in the Arc, whose transcriptional activity is inhibited by T, are targets for the negative feedback regulation of GnRH secretion, whereas KiSS-1 neurons in the anteroventral periventricular nucleus, whose activity is stimulated by T, may mediate other T-dependent processes. PMID- 15831568 TI - Estrogen receptor-beta is critical to granulosa cell differentiation and the ovulatory response to gonadotropins. AB - The process of granulosa cell differentiation that occurs in preovulatory follicles is dependent on FSH but requires augmentation by estradiol. To determine which estrogen receptor (ER) form mediates the effects of estradiol during gonadotropin-induced follicle growth, differentiation, and rupture, we characterized the response of ERalpha- and ERbeta-null mice to gonadotropin induced ovulation. Immature mice were treated with an ovulatory regimen of exogenous gonadotropins and tissues were collected at distinct time points for morphological, biochemical, gene expression, and immunohistochemical analyses. Granulosa cells of ERbeta knockout (ERKO) preovulatory follicles exhibited an attenuated response to FSH-induced differentiation, as evident by reduced aromatase activity and estradiol synthesis, and insufficient expression of LH receptor. As a result, betaERKO ovaries were unable to fully respond to an ovulatory bolus of gonadotropin, leading to a reduced rate of follicle rupture; insufficient induction of prostaglandin-synthase 2 and progesterone receptor; an aberrant increase in aromatase activity and plasma estradiol; and incomplete expansion of the cumulus-oocyte complex. Parallel characterization of alphaERKO females indicated a minimal role for ERalpha in granulosa cell differentiation, ovulation, and the concomitant changes in gene expression, although some abnormalities were revealed. These studies demonstrate that ERbeta-mediated estradiol actions are vital to FSH-induced granulosa cell differentiation; and in the absence of ERbeta, preovulatory follicles are deficient in the necessary cellular organization (i.e. antrum and cumulus oocyte complex), enzymatic activity (i.e. capacity to convert androgen precursor to estradiol), and receptor signaling pathways (i.e. LH receptor) to respond to a gonadotropin surge and expel a healthy oocyte. PMID- 15831569 TI - Origin of cocaine- and amphetamine-regulated transcript-containing axons innervating hypophysiotropic corticotropin-releasing hormone-synthesizing neurons in the rat. AB - Cocaine- and amphetamine-regulated transcript (CART) has stimulatory effects on the hypothalamic-pituitary-adrenal axis through direct effects on hypophysiotropic CRH neurons. Recently CART-containing axons have been demonstrated to densely innervate the hypophysiotropic CRH neurons. Based on the sources of the CART-immunoreactive (IR) innervation of the paraventricular nucleus, the putative origins of these CART-containing fibers include neurons of the hypothalamic arcuate nucleus that coexpress alphaMSH and medullary adrenaline producing neurons. To determine whether these cell groups contribute to the CART innervation of the hypophysiotropic CRH neurons, we performed a quadruple labeling immunofluorescent study using antisera against CRH, CART, alphaMSH, and phenylethanolamine-N-methyl-transferase (PNMT), the latter as a marker for adrenaline. Consistent with previous observations, PNMT- and CART-IR axons densely innervated all CRH neurons, whereas the alphaMSH-IR innervation was sparse. Although approximately 60% of CART-IR varicosities in juxtaposition to CRH neurons cocontained PNMT, only approximately 18% of them were immunopositive for alphaMSH. All alphaMSH-IR boutons and approximately 90% of PNMT-containing varicosities on the surface of CRH neurons were also labeled for CART. The remaining 22% of CART axon varicosities in contact with CRH neurons contained neither alphaMSH nor PNMT. These results indicate that medullary adrenergic/CART neurons are the major source for the CART innervation of CRH neurons in the paraventricular nucleus; however, to a lesser extent the arcuate nucleus also contributes to the CART-IR innervation of these neurons. The observation that nearly 20% of the CART-IR afferents contain neither alphaMSH nor PNMT, however, suggests that additional sources also contribute to the CART-IR input of hypophysiotropic CRH neurons. PMID- 15831570 TI - Regulation of parathyroid hormone-related peptide by estradiol: effect on tumor growth and metastasis in vitro and in vivo. AB - We evaluated the capacity of estradiol (E(2)) to regulate PTHrP production, cell growth, tumor growth, and metastasis to the skeleton in breast cancer. In estrogen receptor (ER)-negative human breast cancer cells, MDA-MB-231, and cells transfected with full-length cDNA encoding ER (S-30), E(2) caused a marked decrease in cell growth and PTHrP production, effects that were abrogated by anti E(2) tamoxifen. E(2) also inhibited PTHrP promoter activity in S-30 cells. For in vivo studies, MDA-MB-231 and S-30 cells were inoculated into the mammary fat pad of female BALB/c nu.nu mice. Animals receiving S-30 cells developed tumors of significantly smaller volume compared with MDA-MB-231 tumor-bearing animals. This change in tumor volume was reversed when S-30 cells were inoculated into ovariectomized (OVX) hosts. Inoculation of MDA-MB-231 cells into the left ventricle resulted in the development of lesions in femora and tibia as determined by x-ray analysis. In contrast, these lesions were significantly smaller in volume and number in animals inoculated with S-30, and this lower incidence was reversed in OVX animals. Bone histological analysis showed that the tumor volume to tissue volume ratio was comparable with that seen by x-ray. Immunohistochemical analysis showed that PTHrP production was inhibited in S-30 group and restored to levels comparable to that seen in MDA-MB-231 tumor-bearing animals when S-30 cells were inoculated in OVX animals. Collectively these studies show that E(2) production is inversely correlated with PTHrP production and that the growth-promoting effect of PTHrP has a direct impact on tumor growth at both nonskeletal and skeletal sites. PMID- 15831571 TI - Calcium has a permissive role in interleukin-1beta-induced c-jun N-terminal kinase activation in insulin-secreting cells. AB - The c-jun N-terminal kinase (JNK) signaling pathway mediates IL-1beta-induced apoptosis in insulin-secreting cells, a mechanism relevant to the destruction of pancreatic beta-cells in type 1 and 2 diabetes. However, the mechanisms that contribute to IL-1beta activation of JNK in beta-cells are largely unknown. In this study, we investigated whether Ca(2+) plays a role for IL-1beta-induced JNK activation. In insulin-secreting rat INS-1 cells cultured in the presence of 11 mm glucose, combined pharmacological blockade of L- and T-type Ca(2+) channels suppressed IL-1beta-induced in vitro phosphorylation of the JNK substrate c-jun and reduced IL-1beta-stimulated activation of JNK1/2 as assessed by immunoblotting. Inhibition of IL-1beta-induced in vitro kinase activity toward c jun after collective L- and T-type Ca(2+) channel blockade was confirmed in primary rat and ob/ob mouse islets and in mouse betaTC3 cells. Ca(2+) influx, specifically via L-type but not T-type channels, contributed to IL-1beta activation of JNK. Activation of p38 and ERK in response to IL-1beta was also dependent on L-type Ca(2+) influx. Membrane depolarization by KCl, exposure to high glucose, treatment with Ca(2+) ionophore A23187, or exposure to thapsigargin, an inhibitor of sarco(endo)plasmic reticulum Ca(2+) ATPase, all caused an amplification of IL-1beta-induced JNK activation in INS-1 cells. Finally, a chelator of intracellular free Ca(2+) [bis-(o-aminophenoxy)-N,N,N',N' tetraacetic acid-acetoxymethyl], an inhibitor of calmodulin (W7), and inhibitors of Ca(2+)/calmodulin-dependent kinase (KN62 and KN93) partially reduced IL-1beta stimulated c-jun phosphorylation in INS-1 or betaTC3 cells. Our data suggest that Ca(2+) plays a permissive role in IL-1beta activation of the JNK signaling pathway in insulin-secreting cells. PMID- 15831572 TI - Stability of neuroendocrine and behavioral responsiveness in aging Fischer 344/Brown-Norway hybrid rats. AB - Aging in rodents and primates is accompanied by changes in hypothalamic-pituitary adrenal (HPA) activity. We examined behavioral and neuroendocrine responses in 3, 15-, and 30-month-old F344/Brown-Norway rats. Basal corticosterone and ACTH levels did not differ with age, although ACTH responses, but not corticosterone responses to restraint stress, were significantly lower in the 30-month-old group relative to 3- and 15-month-old rats. Induction of c-fos mRNA in the paraventricular nucleus from restraint was not affected by age. Furthermore, there was an enhanced sensitivity to dexamethasone suppression in aged animals as evidenced by lesser ACTH and corticosterone release after dexamethasone administration. Evaluation of emotional behaviors in the forced swim test revealed no differences between the age groups. With fear conditioning, aged rats had decreased freeze times relative to middle-aged or young rats. Regression analysis revealed no significant correlations between the behavioral and HPA axis data in any group. Overall, the data suggest that an apparent decrease in pituitary drive is compensated for at the level of the adrenal, resulting in stable patterns of glucocorticoid secretion. The lack of a correlation between HPA axis measures and emotional as well as fear conditioning-related behaviors indicates that corticosteroid dysfunction may not predict age-related behavioral deficits in this aging model. PMID- 15831573 TI - Therapeutic effects of tolvaptan, a potent, selective nonpeptide vasopressin V2 receptor antagonist, in rats with acute and chronic severe hyponatremia. AB - The therapeutic efficacy of tolvaptan (OPC-41061), a potent, selective nonpeptide vasopressin V(2) receptor antagonist, on acute and chronic severe hyponatremia was assessed in rats. Experiments were designed to demonstrate the efficacy of tolvaptan reducing mortality in an acute model, and controlling the extent of serum sodium elevation without causing abnormal animal behavior suggesting neurological symptoms in a chronic model. In the acute model, rats developed rapidly progressive, severe hyponatremia by continuous sc infusion of [deamino Cys(1), D-Arg(8)]-vasopressin (10 ng/h) and forced water-loading (additional 10% initial body weight per day). By d 6, untreated rats had a 47% mortality rate. However, rats treated with repeated oral administrations of tolvaptan (1, 3, and 10 mg/kg) produced dose-dependent aquaresis (i.e. urine volume increased and urine osmolality decreased) that resulted in a gradual increase in plasma sodium concentration. Consequently, tolvaptan treatment reduced mortality and, at higher doses, resulted in no observed deaths. In the gradual model, rats receiving a continuous sc infusion of [deamino-Cys(1), D-Arg(8)]-vasopressin (1 ng/h) combined with a liquid diet were induced to stable, severe hyponatremia (approximately 110 mEq/liter), which lead to increased organ weight and water content. Rats receiving dose titrations of tolvaptan (0.25, 0.5, 1, 2, 4, and 8 mg/kg) increased plasma sodium to healthy levels without causing abnormal animal behavior suggesting neurological symptoms or death, improved hyponatremia-driven increases in wet weight and water content in the organs. Thus, in animal models, analogous to the hyponatremia forms seen in humans, tolvaptan presents exciting therapeutic implications in the management of patients with severe hyponatremia. PMID- 15831574 TI - Improving the calculation of statistical significance in genome-wide scans. AB - Calculations of the significance of results from linkage analysis can be performed by simulation or by theoretical approximation, with or without the assumption of perfect marker information. Here we concentrate on theoretical approximation. Our starting point is the asymptotic approximation formula presented by Lander and Kruglyak (1995, Nature Genetics, 11, 241--247), incorporating the effect of finite marker spacing as suggested by Feingold et al. (1993, American Journal of Human Genetics, 53, 234--251). We consider two distinct ways in which this formula can be improved. Firstly, we present a formula for calculating the crossover rate rho for a pedigree of general structure. For a pedigree set, these values may then be weighted into an overall crossover rate which can be used as input to the original approximation formula. Secondly, the unadjusted p -value formula is based on the assumption of a Normally distributed nonparametric linkage (NPL) score. This leads to conservative or anti-conservative p -values of varying magnitude depending on the pedigree set structure. We adjust for non-Normality by calculating the marginal distribution of the NPL score under the null hypothesis of no linkage with an arbitrarily small error. The NPL score is then transformed to have a marginal standard Normal distribution and the transformed maximal NPL score, together with a slightly corrected value of the overall crossover rate, is inserted into the original formula in order to calculate the p -value. We use pedigrees of seven different structures to compare the performance of our suggested approximation formula to the original approximation formula, with and without skewness correction, and to results found by simulation. We also apply the suggested formula to two real pedigree set structure examples. Our method generally seems to provide improved behavior, especially for pedigree sets which show clear departure from Normality, in relation to the competing approximations. PMID- 15831575 TI - Tree-augmented Cox proportional hazards models. AB - We study a hybrid model that combines Cox proportional hazards regression with tree-structured modeling. The main idea is to use step functions, provided by a tree structure, to 'augment' Cox (1972) proportional hazards models. The proposed model not only provides a natural assessment of the adequacy of the Cox proportional hazards model but also improves its model fitting without loss of interpretability. Both simulations and an empirical example are provided to illustrate the use of the proposed method. PMID- 15831576 TI - Exploratory analysis of longitudinal trials with staggered intervention times. AB - Longitudinal trials involving surgical interventions commonly have subject specific intervention times, due to constraints on the availability of surgeons and operating theatres. Moreover, the intervention often effects a discontinuous change in the mean response. We propose a nonparametric estimator for the mean response profile of longitudinal data with staggered intervention times and a discontinuity at the times of intervention, as an exploratory tool to assist the formulation of a suitable parametric model. We use an adaptation of the standard generalized additive model algorithm for estimation, with smoothing constants chosen by a cross-validation criterion. We illustrate the method using longitudinal data from a trial to assess the effect of lung resection surgery in the treatment of emphysema patients. PMID- 15831577 TI - Reduced rank proportional hazards model for competing risks. AB - Competing events concerning individual subjects are of interest in many medical studies. For example, leukemia-free patients surviving a bone marrow transplant are at risk of developing acute or chronic graft-versus-host disease, or they might develop infections. In this situation, competing risks models provide a natural framework to describe the disease. When incorporating covariates influencing the transition intensities, an obvious approach is to use Cox's proportional hazards model for each of the transitions separately. A practical problem then is how to deal with the abundance of regression parameters. Our objective is to describe the competing risks model in fewer parameters, both in order to avoid imprecise estimation in transitions with rare events and in order to facilitate interpretation of these estimates. Suppose that the regression parameters are gathered into a p x K matrix B, with p and K as the number of covariates and transitions, respectively. We propose the use of reduced rank models, where B is required to be of lower rank R, smaller than both p and K. One way to achieve this is to write B = AGamma(intercal) with A and Gamma matrices of dimensions p x R and K x R, respectively. We shall outline an algorithm to obtain estimates and their standard errors in a reduced rank proportional hazards model for competing risks and illustrate the approach on a competing risks model applied to 8966 leukemia patients from the European Group for Blood and Marrow Transplantation. PMID- 15831579 TI - A transformation approach for incorporating monotone or unimodal constraints. AB - Samples of curves are collected in many applications, including studies of reproductive hormone levels in the menstrual cycle. Many approaches have been proposed for correlated functional data of this type, including smoothing spline methods and other flexible parametric modeling strategies. In many cases, the underlying biological processes involved restrict the curve to follow a particular shape. For example, progesterone levels in healthy women increase during the menstrual cycle to a peak achieved at random location with decreases thereafter. Reproductive epidemiologists are interested in studying the distribution of the peak and the trajectory for women in different groups. Motivated by this application, we propose a simple approach for restricting each woman's mean trajectory to follow an umbrella shape. An unconstrained hierarchical Bayesian model is used to characterize the data, and draws from the posterior distribution obtained using a Gibbs sampler are then mapped to the constrained space. Inferences are based on the resulting quasi-posterior distribution for the peak and individual woman trajectories. The methods are applied to a study comparing progesterone trajectories for conception and nonconception cycles. PMID- 15831578 TI - Accuracy of MSI testing in predicting germline mutations of MSH2 and MLH1: a case study in Bayesian meta-analysis of diagnostic tests without a gold standard. AB - Microsatellite instability (MSI) testing is a common screening procedure used to identify families that may harbor mutations of a mismatch repair (MMR) gene and therefore may be at high risk for hereditary colorectal cancer. A reliable estimate of sensitivity and specificity of MSI for detecting germline mutations of MMR genes is critical in genetic counseling and colorectal cancer prevention. Several studies published results of both MSI and mutation analysis on the same subjects. In this article we perform a meta-analysis of these studies and obtain estimates that can be directly used in counseling and screening. In particular, we estimate the sensitivity of MSI for detecting mutations of MSH2 and MLH1 to be 0.81 (0.73-0.89). Statistically, challenges arise from the following: (a) traditional mutation analysis methods used in these studies cannot be considered a gold standard for the identification of mutations; (b) studies are heterogeneous in both the design and the populations considered; and (c) studies may include different patterns of missing data resulting from partial testing of the populations sampled. We address these challenges in the context of a Bayesian meta-analytic implementation of the Hui-Walter design, tailored to account for various forms of incomplete data. Posterior inference is handled via a Gibbs sampler. PMID- 15831580 TI - A general model for detecting genetic determinants underlying longitudinal traits with unequally spaced measurements and nonstationary covariance structure. AB - A mixture model for determining quantitative trait loci (QTL) affecting growth trajectories has been proposed in the literature. In this article, we extend this model to a more general situation in which longitudinal traits for each subject are measured at unequally spaced time intervals, different subjects have different measurement patterns, and the residual correlation within subjects is nonstationary. We derive an EM-simplex hybrid algorithm to estimate the allele frequencies, Hardy-Weinberg disequilibrium, and linkage disequilibrium between QTL in the original population and parameters contained in the growth equation and in the covariance structure. A worked example of head circumference growth in 145 children is used to validate our extended model. A simulation study is performed to examine the statistical properties of the parameter estimation obtained from this example. Finally, we discuss the implications and extensions of our model for detecting QTL that affect growth trajectories. PMID- 15831581 TI - Analysis of clustered recurrent event data with application to hospitalization rates among renal failure patients. AB - End-stage renal disease (commonly referred to as renal failure) is of increasing concern in the United States and many countries worldwide. Incidence rates have increased, while the supply of donor organs has not kept pace with the demand. Although renal transplantation has generally been shown to be superior to dialysis with respect to mortality, very little research has been directed towards comparing transplant and wait-list patients with respect to morbidity. Using national data from the Scientific Registry of Transplant Recipients, we compare transplant and wait-list hospitalization rates. Hospitalizations are subject to two levels of dependence. In addition to the dependence among within patient events, patients are also clustered by listing center. We propose two marginal methods to analyze such clustered recurrent event data; the first model postulates a common baseline event rate, while the second features cluster specific baseline rates. Our results indicate that kidney transplantation offers a significant decrease in hospitalization, but that the effect is negated by a waiting time (until transplant) of more than 2 years. Moreover, graft failure (GF) results in a significant increase in the hospitalization rate which is greatest in the first month post-GF, but remains significantly elevated up to 4 years later. We also compare results from the proposed models to those based on a frailty model, with the various methods compared and contrasted. PMID- 15831582 TI - On the relation between initial value and slope. AB - Suppose measurements of a particular feature are collected at baseline and at a number of subsequent time points and that for each individual there is a roughly linear trend in time. This paper takes three approaches to testing whether there is a relation between the initial value and the slope. It also considers whether the initial value for an individual is a useful predictor of the slope for that individual. The problems are formulated in terms of regression models with random coefficients. The solutions are illustrated using data from an observational study of clinical correlates of disability and progression in Huntington's disease. PMID- 15831583 TI - BGX: a fully Bayesian integrated approach to the analysis of Affymetrix GeneChip data. AB - We present Bayesian hierarchical models for the analysis of Affymetrix GeneChip data. The approach we take differs from other available approaches in two fundamental aspects. Firstly, we aim to integrate all processing steps of the raw data in a common statistically coherent framework, allowing all components and thus associated errors to be considered simultaneously. Secondly, inference is based on the full posterior distribution of gene expression indices and derived quantities, such as fold changes or ranks, rather than on single point estimates. Measures of uncertainty on these quantities are thus available. The models presented represent the first building block for integrated Bayesian Analysis of Affymetrix GeneChip data: the models take into account additive as well as multiplicative error, gene expression levels are estimated using perfect match and a fraction of mismatch probes and are modeled on the log scale. Background correction is incorporated by modeling true signal and cross-hybridization explicitly, and a need for further normalization is considerably reduced by allowing for array-specific distributions of nonspecific hybridization. When replicate arrays are available for a condition, posterior distributions of condition-specific gene expression indices are estimated directly, by a simultaneous consideration of replicate probe sets, avoiding averaging over estimates obtained from individual replicate arrays. The performance of the Bayesian model is compared to that of standard available point estimate methods on subsets of the well known GeneLogic and Affymetrix spike-in data. The Bayesian model is found to perform well and the integrated procedure presented appears to hold considerable promise for further development. PMID- 15831584 TI - Failure time analysis of HIV vaccine effects on viral load and antiretroviral therapy initiation. AB - The world's first efficacy trial of a preventive HIV vaccine was completed in 2003. Study participants who became HIV infected were followed for 2 years and monitored for HIV viral load and initiation of antiretroviral therapy (ART). In order to determine if vaccination may have altered HIV progression in persons who acquired HIV, a pre-specified objective was to compare the time until a composite endpoint between the vaccine and placebo arms, where the composite endpoint is the first event of ART initiation or viral failure (HIV viral load exceeds a threshold x(vl) copies/ml). Specifically, with vaccine efficacy, VE(tau, x(vl)), defined as one minus the ratio (vaccine/placebo) of the cumulative probability of the composite endpoint (with failure threshold x(vl)) occurring by tau months, the aim was to estimate the four parameters {VE(tau, x(vl)): x(vl) is an element of {1500, 10,000, 20,000, 55,000} copies/ml} with simultaneous 95% confidence bands. A Gaussian multipliers simulation method is devised for constructing confidence bands for VE(tau, x(vl)) with x(vl) spanning multiple discrete values or a continuous range. The new method is evaluated in simulations and is applied to the vaccine trial data set. PMID- 15831585 TI - Direct activation of fission yeast adenylate cyclase by the Gpa2 Galpha of the glucose signaling pathway. AB - G protein-mediated signaling is implicated in yeast and fungal cAMP pathways. By two-hybrid screens and pull-down experiments, we show that the fission yeast Gpa2 Galpha binds an N-terminal domain of adenylate cyclase, comprising a moderately conserved sequence within a region otherwise poorly related to other fungal adenylate cyclases. Overexpressing this domain in yeast perturbs cAMP signaling, which is restored by Gpa2 coexpression. Mutations affecting this domain, over 1,100 residues from the catalytic domain, alter glucose-triggered cAMP signaling. This is evidence for direct activation of adenylate cyclase by a fungal G protein and suggests a distinct activation mechanism from that of mammals. PMID- 15831586 TI - Effector CD8+ T cells recovered from an influenza pneumonia differentiate to a state of focused gene expression. AB - The restriction of influenza A virus replication to mouse respiratory epithelium means that this host response is anatomically compartmentalized, on the one hand, to sites of T cell stimulation and proliferation in the secondary lymphoid tissue and, on the other hand, to the site of effector T cell function and pathology in the pneumonic lung. Thus, it is hardly surprising that virus-specific CD8(+) T cells recovered by bronchoalveolar lavage (BAL) from the infected respiratory tract seem more "activated" in terms of both cytolytic activity and cytokine production than those cells isolated from the spleen. The present analysis uses Affymetrix microarray technology to compare profiles of gene expression in these two lineage-related, yet anatomically separate, lymphocyte populations. Ninety differentially expressed genes were identified for influenza-specific CD8(+)D(b)NP(366)(+) T cells obtained directly ex vivo by BAL or spleen disruption, with nine genes being further analyzed by quantitative, real-time PCR at the population level. Integrin alphaE, for example, was shown by Affymetrix and real-time mRNA analyses and then by single-cell PCR and protein staining to be present at a much higher prevalence on the BAL CD8(+)D(b)NP(366)(+) set. The unpredicted finding, however, was that mRNA expression for 75% of the 90 genes was lower in T cells from the BAL than from the spleen. Apparently, the localization of virus-specific CD8(+) T cells to the site of virus-induced pathology is associated with a narrowing, or "focusing," of gene expression that favors enhanced effector function in the damaged, "high-antigen load" environment of the pneumonic lung. PMID- 15831587 TI - Glutamate induces the rapid formation of spine head protrusions in hippocampal slice cultures. AB - Synaptic plasticity at neuronal connections has been well characterized functionally by using electrophysiological approaches, but the structural basis for this phenomenon remains controversial. We have studied the dynamic interactions between presynaptic and postsynaptic structures labeled with FM 4-64 and a membrane-targeted GFP, respectively, in hippocampal slices. Under conditions of reduced neuronal activity (1 muM tetrodotoxin), we observed extension of glutamate receptor-dependent processes from dendritic spines of CA1 pyramidal cells to presynaptic boutons. The formation of these spine head protrusions is blocked by alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists and by agents that reduce the release of glutamate from presynaptic terminals. Moreover, spine head protrusions form in response to exogenously applied glutamate, with clear directionality toward the glutamate electrode. Our results suggest that spontaneously released glutamate is sufficient to activate nearby spines, which can then lead to the growth of new postsynaptic processes connecting to a presynaptic site. Spines thus can compare their recent history with that of neighboring synapses and modify local connectivity accordingly. PMID- 15831588 TI - Semipermeable lipid bilayers exhibit diastereoselectivity favoring ribose. AB - Nutrient uptake by a primitive cell would have been limited by the permeability characteristics of its membrane. We measured the permeabilities of model protocellular membranes to water, five of the six pentoses, and selected aldohexoses, ketohexoses, and three to six carbon alditols by following volume changes of vesicles after the addition of solute to the external medium. Solute hydrophobicities correlated poorly with permeability coefficients within one structural class of compounds. The permeability coefficients of diastereomeric sugars differed by as much as a factor of 10, with ribose being the most permeable aldopentose. Flexible alditols and sugars, sugars biased toward or restricted to furanose forms, and sugars having anomers with hydrophobic faces permeated more quickly than compounds lacking these features. Among the aldopentoses, only ribose possesses all of these properties. Ribose permeated both fatty acid and phospholipid membranes more rapidly than the other aldopentoses or hexoses. The enhanced permeability conferred by the unique conformational preferences of ribose would have allowed faster assimilation of ribose by primitive cells as they passively absorbed materials from the environment. The kinetic advantage of ribose over the other aldopentoses in crossing membranes may therefore have been one factor that facilitated the emergence of the RNA world. PMID- 15831589 TI - Supramolecular blueprint approach to metal-coordinated capsules. AB - An important problem in designing any large network is the assembly of systems that are resilient to change. From a chemical point of view, an analogy can be used where one requires supramolecular assemblies to maintain their dimensionality combined with limited structural perturbation in response to variation in its intermolecular framework. The identification of hydrogen-bonded framework patterns within experimentally known supramolecular assemblies that are structurally robust to disruption and selective hydrogen substitution are envisioned to act as a supramolecular blueprint or template for metal-ion retroinsertion. Here, we report the formation of a large neutral discrete pseudo spherical coordination capsule assembled from 6 pyrogallol[4]arene ligands and 24 Cu(II) metal ions. Amazingly, this coordination capsule is structurally analogous to its hydrogen-bonded counterpart. This result shows a robust ability of pyrogallol[4]arene molecules to self-assemble into large hexameric cage structures from either the hydrogen-bonding or metal-ligand coordination process. The identification of robust supramolecular assemblies that conserve their structure in response to interchangeability between hydrogen-bonded networks for metal coordination, or inversely, represents an important advancement in supramolecular design. PMID- 15831590 TI - Ab initio molecular dynamics and quasichemical study of H+(aq). AB - The excess proton in water, H(+)(aq), plays a fundamental role in aqueous solution chemistry. Its solution thermodynamic properties are essential to molecular descriptions of that chemistry and for validation of dynamical calculations. Within the quasichemical theory of solutions those thermodynamic properties are conditional on recognizing underlying solution structures. The quasichemical treatment identifies H(3)O(+) and H(2)O(5)(+) as natural inner shell complexes, corresponding to the cases of n = 1, 2 water molecule ligands, respectively, of a distinguished H(+) ion. A quantum-mechanical treatment of the inner-shell complex with both a dielectric continuum and a classical molecular dynamics treatment of the outer-shell contribution identifies the latter case (the Zundel complex) as the more numerous species. Ab initio molecular dynamics simulations, with two different electron density functionals, suggest a preponderance of Zundel-like structures, but a symmetrical ideal Zundel cation is not observed. PMID- 15831591 TI - A mutation in T7 RNA polymerase that facilitates promoter clearance. AB - Like multisubunit RNA polymerases (RNAPs), T7 RNAP frequently releases its transcript over the initial 8-12 transcribed nucleotides, when it still contacts the promoter. This abortive cycling, which is most prominent with initial sequences that deviate from those of T7 late genes, eventually compromises productive transcription. Starting from an in vivo situation where transcription of a target gene by T7 RNAP is virtually abolished because of extensive abortive cycling, we have selected a mutation in RNAP that restores target gene expression. In vitro, this mutation (P266L) weakens promoter binding but markedly reduces abortive cycling over a variety of initial sequences by stabilizing the transcription complex at nucleotides 5-8. Other substitutions of P266 have similar effects. X-ray data show that during the transition from initial to elongation complex, the N-terminal region undergoes a major structural switch of which P266 constitutes one of the hinges. How the mutation might facilitate this switch is tentatively discussed. On the practical side, the mutation can significantly improve in vitro transcription, particularly from templates carrying unfavorable initial sequences. PMID- 15831592 TI - Systematic assessment of atypical deletions reveals genotype-phenotype correlation in 22q11.2. AB - INTRODUCTION: Clinical variability associated with the common 22q11.2 microdeletion is well known, and has led to a broad application of FISH diagnostics with probes for loci TUPLE1 or D22S75 (N25), although, rarely reported atypical deletions associated with the same phenotypic spectrum would not be discovered by these probes. As most types of 22q11.2 deletions occur between low copy repeats within the region (LCR22), we assumed that atypical deletions should be more common than has been reported. To address this question and the possibility of a deletion size related genotype-phenotype correlation, we systematically assessed the frequency of typical and atypical 22q11.2 deletions in a large cohort of patients. METHODS: We used a set of 10 fluorescent in situ hybridisation (FISH) DNA probes, capable of detecting all reported and hypothetical deletions between the LCR22, and analysed 350 patients. Deletion sizes in atypical deletions were established by use of further FISH probes. Frequency of certain atypical deletions was analysed in controls by FISH and quantitative PCR. RESULTS: Patients with conotruncal heart defects (ctCHD) and with typical VCFS phenotype showed the common 3 Mb or nested 1.5 Mb deletions (in 18.5% and 78.6%, respectively), but no atypical deletion, while 5% (3/63) of patients with a mildly suggestive, atypical phenotype showed atypical distal deletions, which were not detected in patients with mental retardation of unknown origin or in healthy controls. DISCUSSION: These statistically significant differences demonstrate that atypical distal 22q11.2 deletions are very uncommon in patients with ctCHDs, while atypical congenital heart defects and mild dysmorphism are recognisable feature of atypical distal deletions. Further phenotype-genotype analysis disclosed association of significant developmental delay with the distal part of the common deletion region, and choanal atresia and atypical CHDs with the adjacent distal deletion region. PMID- 15831593 TI - Cleft lip/palate and CDH1/E-cadherin mutations in families with hereditary diffuse gastric cancer. AB - We report the association of CDH1/E-cadherin mutations with cleft lip, with or without cleft palate (CLP), in two families with hereditary diffuse gastric cancer (HDGC). In each family, the CDH1 mutation was a splicing mutation generating aberrant transcripts with an in-frame deletion, removing the extracellular cadherin repeat domains involved in cell-cell adhesion. Such transcripts might encode mutant proteins with trans-dominant negative effects. We found that CDH1 is highly expressed at 4 and 5 weeks in the frontonasal prominence, and at 6 weeks in the lateral and medial nasal prominences of human embryos, and is therefore expressed during the critical stages of lip and palate development. These findings suggest that alteration of the E-cadherin pathway can contribute to human clefting. PMID- 15831594 TI - Methodological issues in longitudinal studies: vestibular schwannoma growth rates in neurofibromatosis 2. AB - Four longitudinal studies of vestibular schwannoma (VS) growth rates in neurofibromatosis 2 (NF2) have yielded very different results on the relationship of VS growth rates to age. The studies had different patient eligibility criteria, indices of VS growth rates, VS volumetric methods, and sample sizes. We reanalysed data from two of the longitudinal studies and used data from the population based United Kingdom NF2 Registry to determine the most likely reason for the different results and the actual relationship of VS growth rates to age. We found that the eligibility criterion in one study caused selection bias for slower growing VS. The proper interpretation of the results from the four studies is that VS growth rates in NF2 are highly variable but tend to decrease with increasing age. Clinical trials for VS in NF2 should focus on younger patients because VS growth rates tend to decrease with increasing age, and because faster growing VS are more likely to be excised with increasing age than slower growing VS. PMID- 15831597 TI - Myocardial viability assessed by dobutamine stress echocardiography predicts reduced mortality early after acute myocardial infarction: determining the risk of events after myocardial infarction (DREAM) study. AB - OBJECTIVE: To establish further the role of dobutamine stress echocardiography (DSE) in prognostication of outcome early after acute myocardial infarction (AMI) METHODS: Consecutive patients presenting with AMI were screened for inclusion into the study. 212 stable consenting patients underwent DSE a mean (SD) of 4.8 (1.5) days after AMI. Patients were then followed up for 803 (297) days. RESULTS: The mean (SD) resting systolic wall thickening index (SWTI) was 1.6 (0.4), 44% patients had evidence of viability at low dose, and 38% had evidence of ischaemia. During the follow up period 27 (13%) patients died and 16 (8%) had a non-fatal AMI. Independent predictors of both mortality and combined mortality and non-fatal AMI were age (hazard ratio (HR) 1.04/year, p = 0.01, and HR 1.03/year, p = 0.04, respectively) and SWTI at low dose (HR 3.6, p < 0.01, and HR 2.5, p = 0.02, respectively). Low dose DSE provided incremental information over clinical and resting left ventricular function data for predicting death and non fatal AMI. For patients who were not revascularised SWTI at peak dose dobutamine was the only independent predictor of mortality. CONCLUSION: DSE is a powerful predictor of outcome in stable survivors of AMI. The presence of myocardial viability has a positive impact on survival. PMID- 15831596 TI - Primary percutaneous coronary intervention versus thrombolytic treatment: long term follow up according to infarct location. AB - OBJECTIVES: To study the clinical significance of infarct location during long term follow up in a trial comparing thrombolysis with primary angioplasty. DESIGN: Retrospective longitudinal cohort analysis of prospectively entered data. SETTING: Patients with acute ST elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI). PATIENTS: In the Zwolle trial 395 patients with acute STEMI were randomly assigned to intravenous streptokinase or PCI. MAIN OUTCOME MEASURES: Survival according to infarct location and treatment after 8 (2) years of follow up. RESULTS: 105 patients died: 63 patients in the streptokinase group and 42 patients in the primary PCI group (relative risk (RR) 1.6, 95% confidence interval (CI) 1.0 to 2.6; p = 0.03). In patients with non-anterior STEMI there was no difference in mortality between streptokinase and PCI treated patients (RR 1.1, 95% CI 0.6 to 2.1; p = 0.68) but the streptokinase group had significantly more major adverse cardiac events (MACE) than the PCI group (RR 2.1, 95% CI 1.2 to 3.6). The number needed to treat to prevent one MACE was four. In patients with anterior STEMI, mortality was higher in the streptokinase group than in the PCI group (RR 2.7, 95% CI 1.4 to 5.5; p = 0.004). The number needed to treat to prevent one death was five. Kaplan-Meier analysis confirmed the benefits of primary angioplasty in the first year and showed additional benefit of PCI compared with streptokinase between 1-8 years after the acute event. CONCLUSIONS: Patients with anterior STEMI have better long term survival when treated with PCI than with streptokinase. In patients alive one year after the acute event, PCI confers a significant additional survival benefit, probably due to better preserved residual left ventricular function. PMID- 15831595 TI - Whole genomewide linkage screen for neural tube defects reveals regions of interest on chromosomes 7 and 10. AB - Neural tube defects (NTDs) are the second most common birth defects (1 in 1000 live births) in the world. Periconceptional maternal folate supplementation reduces NTD risk by 50-70%; however, studies of folate related and other developmental genes in humans have failed to definitively identify a major causal gene for NTD. The aetiology of NTDs remains unknown and both genetic and environmental factors are implicated. We present findings from a microsatellite based screen of 44 multiplex pedigrees ascertained through the NTD Collaborative Group. For the linkage analysis, we defined our phenotype narrowly by considering individuals with a lumbosacral level myelomeningocele as affected, then we expanded the phenotype to include all types of NTDs. Two point parametric analyses were performed using VITESSE and HOMOG. Multipoint parametric and nonparametric analyses were performed using ALLEGRO. Initial results identified chromosomes 7 and 10, both with maximum parametric multipoint lod scores (Mlod) >2.0. Chromosome 7 produced the highest score in the 24 cM interval between D7S3056 and D7S3051 (parametric Mlod 2.45; nonparametric Mlod 1.89). Further investigation demonstrated that results on chromosome 7 were being primarily driven by a single large pedigree (parametric Mlod 2.40). When this family was removed from analysis, chromosome 10 was the most interesting region, with a peak Mlod of 2.25 at D10S1731. Based on mouse human synteny, two candidate genes (Meox2, Twist1) were identified on chromosome 7. A review of public databases revealed three biologically plausible candidates (FGFR2, GFRA1, Pax2) on chromosome 10. The results from this screen provide valuable positional data for prioritisation of candidate gene assessment in future studies of NTDs. PMID- 15831598 TI - Permanent junctional reciprocating tachycardia in children: a multicentre study on clinical profile and outcome. AB - OBJECTIVES: To investigate the clinical profile, natural history, and optimal management of persistent or permanent junctional reciprocating tachycardia (PJRT) in children. METHODS AND RESULTS: 85 patients meeting the ECG criteria for PJRT were enrolled in a retrospective multicentre study. Age at diagnosis varied from birth to 20 years (median 3 months). Follow up ranged from 0.1 to 26.0 (median 8.2) years. At the time of referral, 24 of 85 patients (28%) had congestive heart failure that was resolved with medical treatment in all patients. Eighty three patients received drug treatment initially. Amiodarone and verapamil were the most effective with a success rate of 84-94% alone or in association with digoxin. Radiofrequency ablation of the accessory pathway was performed in 18 patients. There was a trend for a relation between age at ablation and the result of the procedure, failures being more common in younger patients (three of six procedures in younger and 15 of 18 in older children were successful; p = 0.14). Two patients with persistent left ventricular dysfunction on echocardiography but with no symptoms of congestive heart failure died suddenly one month and three years after diagnosis. PJRT resolved spontaneously in 19 patients (22%). Age at diagnosis of PJRT was not a predictor of spontaneous resolution. CONCLUSIONS: PJRT is a potentially lethal arrhythmia in children with tachycardia induced cardiomyopathy. Spontaneous resolution of tachycardia is not uncommon. Antiarrhythmic treatment is often effective. Radiofrequency ablation should be performed in older children or when rate is not controlled, especially in patients with persistent left ventricular dysfunction. PMID- 15831599 TI - Cost effectiveness of drug eluting coronary artery stenting in a UK setting: cost utility study. AB - OBJECTIVE: To assess the cost effectiveness of drug eluting stents (DES) compared with conventional stents for treatment of symptomatic coronary artery disease in the UK. DESIGN: Cost-utility analysis of audit based patient subgroups by means of a simple economic model. SETTING: Tertiary care. PARTICIPANTS: 12 month audit data for 2884 patients receiving percutaneous coronary intervention with stenting at the Cardiothoracic Centre Liverpool between January 2000 and December 2002. MAIN OUTCOME MEASURES: Risk of repeat revascularisation within 12 months of index procedure and reduction in risk from use of DES. Economic modelling was used to estimate the cost-utility ratio and threshold price premium. RESULTS: Four factors were identified for patients undergoing elective surgery (n = 1951) and two for non-elective surgery (n = 933) to predict risk of repeat revascularisation within 12 months. Most patients fell within the subgroup with lowest risk (57% of the elective surgery group with 5.6% risk and 91% of the non elective surgery group with 9.9% risk). Modelled cost-utility ratios were acceptable for only one group of high risk patients undergoing non-elective surgery (only one patient in audit data). Restricting the number of DES for each patient improved results marginally: 4% of stents could then be drug eluting on economic grounds. The threshold price premium justifying 90% substitution of conventional stents was estimated to be 112 pound sterling (212 USD, 162 pound sterling) (sirolimus stents) or 89 pound sterling (167 USD, 130 pound sterling) (paclitaxel stents). CONCLUSIONS: At current UK prices, DES are not cost effective compared with conventional stents except for a small minority of patients. Although the technology is clearly effective, general substitution is not justified unless the price premium falls substantially. PMID- 15831600 TI - The British Cardiac Society Working Group definition of myocardial infarction: implications for practice. AB - OBJECTIVE: To assess the impact on observed mortality of the British Cardiac Society (BCS) definition of myocardial infarction (MI) in 11 UK hospitals. DESIGN: Prospective observational registry. SETTING: 11 adjacent hospitals in the West Yorkshire region. PATIENTS: 2484 patients with the acute coronary syndrome (ACS) were identified during a six month period (28 April to 28 October 2003). Demographic, clinical, and treatment variables were collected on all patients. Deaths were monitored through the Office of National Statistics. Patients were categorised into three groups according to the BCS definition of MI: ACS with unstable angina (UA), ACS with myocyte necrosis, and ACS with clinical MI. RESULTS: 30 day mortality was 4.5%, 10.4%, and 12.9% (p < 0.001) in the ACS with UA, ACS with myocyte necrosis, and ACS with clinical MI groups, respectively. At six months the mortality for patients in the groups ACS with clinical MI and ACS with myocyte necrosis was similar (19.2% v 18.7%), being higher than for ACS with UA (8.6%). Same admission percutaneous coronary intervention was similar in groups with clinical MI and myocyte necrosis (11.1% v 10.7%, respectively) as was coronary artery bypass grafting (2.6% v 2.7%, respectively). However, these two groups differed significantly in the prescribing of secondary prevention (aspirin, 79% v 69%; statins, 80% v 68%; beta blockers, 66% v 53%; and angiotensin converting enzyme inhibitors, 65% v 53%; p < 0.001). CONCLUSIONS: At 30 days the new BCS categories for MI predict three distinct outcomes. However, within a contemporary UK population this was no longer apparent at six months, as mortality for patients with ACS with myocyte necrosis had risen to the same level as those for patients with ACS with clinical MI. One possible explanation for this is the apparent under use of drugs known to improve prognosis after traditionally defined MI. PMID- 15831601 TI - Pathophysiology of heart failure following myocardial infarction. AB - The structural and functional abnormalities that lead to cardiac death are coronary artery disease and left ventricular abnormalities related to remodelling (left ventricular hypertrophy, left ventricular systolic dysfunction, and left ventricular fibrosis). Aldosterone adversely affects all of these processes. It produces both a vasculopathy and left ventricular dysfunction and fibrosis. Endothelial dysfunction in the coronary arteries can lead to acute coronary events. Left ventricular dysfunction will cause the progression of heart failure, and left ventricular fibrosis and dysfunction provide an arrhythmic substrate. The combination of acute coronary events and arrhythmias can lead to sudden cardiac deaths, while acceleration of the heart failure disease process can lead to deaths from progressive heart failure. The increased understanding of the mechanistic role of aldosterone in cardiovascular disease provides a rationale for the positive results that have been seen in clinical trials of aldosterone blockade. PMID- 15831602 TI - Treatments that improve outcome in the patient with heart failure, left ventricular systolic dysfunction, or both after acute myocardial infarction. AB - Patients with heart failure, left ventricular systolic dysfunction, or both, after acute myocardial infarction have a poor prognosis. It is important to focus treatment on this high risk group to reduce the persistently high morbidity and mortality after acute myocardial infarction. As in chronic heart failure, there is now good evidence that inhibition of the renin-angiotensin-aldosterone system and sympathetic nervous system, with the appropriate drugs, can reduce morbidity and mortality. In addition to angiotensin converting enzyme inhibitors, angiotensin receptor blockers, and beta blockers, the aldosterone blocker eplerenone has now been shown to be effective in reducing adverse outcomes. PMID- 15831603 TI - From CCU to CHF: bridging the treatment gap. PMID- 15831604 TI - What can we learn from Europe? AB - The European Society of Cardiology has produced guidelines for the treatment of acute myocardial infarction as well as for chronic heart failure and for the use of beta blockers and angiotensin converting enzyme inhibitors. These documents provide clear evidence and strength of recommendations for the secondary prevention of complications after a myocardial infarction. The identification of heart failure and left ventricular systolic dysfunction are important risk factors in this context. The use of secondary prevention treatments in Europe has been evaluated in several surveys. The use of treatments varies across the participating countries and evidence based therapies in general are under utilised. Various approaches have been taken to disseminate evidence based secondary prevention. Experience from the Italian BRING-UP collaboration illustrates how the use of beta blockers can be increased. Similarly, the Swedish RIKS-HIA registry of acute myocardial infarction has increased the use of secondary preventive treatments. PMID- 15831605 TI - Lessons from the management of chronic heart failure. AB - In seeking to implement evidence based medicine for the patient with heart failure occurring after a myocardial infarction (MI), much can be learnt from the long road to delivery of best care for the patient with chronic heart failure (CHF) caused by left ventricular systolic dysfunction. Both patient groups are part of the same cardiovascular continuum. A mass of evidence has accrued for the beneficial effects of angiotensin converting enzyme inhibitors, beta blockers, and aldosterone antagonists on both morbidity and mortality across a wide spectrum of patient severity. This evidence has informed the development of management guidelines, although registry data showed that uptake of treatments remained low, leading to research focused on how heart failure care could be delivered more effectively. This has resulted in a range of heart failure management programmes, many of which have been shown to reduce hospital admission rates and to improve adherence with treatments. Multidisciplinary heart failure management programmes that span primary and secondary care are now considered a routine "standard" to be aspired to in delivering effective CHF care. Applying such an approach to the care of the post-MI heart failure patient should be equally important. PMID- 15831606 TI - Lessons from the management of acute myocardial infarction. AB - The National Service Framework for coronary heart disease set a number of challenging targets for the care of patients following an acute myocardial infarction. The Myocardial Infarction National Audit Project (MINAP) was devised to monitor progress and has been notably successful in winning professional support and participation and helping trusts to meet these targets. The new challenge is in translating this success to other areas of medicine. Heart failure is one such area, although it poses a number of difficulties relating primarily to disease definition and the definition of a successful outcome. MINAP was overseen by a multidisciplinary group of stakeholders, including patient organisations, and was project managed by a professionally led team at the Royal College of Physicians. Successful projects must retain confidence of all stakeholders and in part this depends on ensuring that timelines are met. Central monitoring of returns and anticipation of problems has been an important component of data completeness and quality. Next day updates to those collecting the data and more detailed quarterly reports for clinicians and chief executives within days of quarter end have been vital. Change depends on clinicians and managers working together. But most importantly, the attention to detail outlined above means the data have been believed and the resulting change for patients has been remarkable. PMID- 15831607 TI - Heart failure post-myocardial infarction: a review of the issues. AB - In most patients with heart failure due to left ventricular systolic dysfunction, the underlying cause is coronary heart disease. To reduce progression to heart failure in a patient with acute myocardial infarction, it is important to achieve the earliest possible reperfusion, whether by thrombolysis or primary percutaneous coronary intervention. Every patient with acute myocardial infarction should have an assessment of their left ventricular function, the potential for reversibility should be considered, and reversible ischaemia should be identified. Left ventricular dysfunction does not only occur with ST segment elevation myocardial infarction but is also commonly associated with non-ST segment elevation myocardial infarction. Secondary prevention is crucial and this requires long term commitment by the patient and the health care system. Heart failure and left ventricular dysfunction are treatable but require a multidisciplinary, integrated network approach. PMID- 15831609 TI - Bridging the treatment gap: the secondary care perspective. AB - Protocols and integrated care pathways can be valuable in the hospital care of patients with heart failure or left ventricular systolic dysfunction after acute myocardial infarction. A designated member of staff, often a specialist nurse, must be responsible for identifying patients suitable for management by the protocol and for ensuring that the protocol is adhered to. A new training scheme for "limited echocardiography" might enable specialist nurses to investigate left ventricular function within the first 24 hours of admission. Patients should be discharged from hospital as soon as they are out of danger. At present, they are often kept in hospital for process reasons. A "continuing care" clinic run by a specialist nurse, where patients can be seen daily after discharge until they are stabilised, is one way of bridging the gap between secondary and primary care. Communication between secondary and primary care needs to improve and same day discharge summaries are essential. PMID- 15831610 TI - Bridging the treatment gap: the primary care perspective. AB - The Darlington heart failure service model, part of the South Durham Heart Failure Network, was devised to overcome barriers to accurate diagnosis and effective management of heart failure. It involves rapid diagnosis of left ventricular systolic dysfunction (LVSD) and ongoing heart failure management. A weekly one stop diagnostic clinic, run by a general practitioner (GP) specialist and a heart failure nurse, is jointly funded by the primary care trust and the South Durham NHS Trust. If LVSD is confirmed, a management plan is formulated which includes patient education and initiation of evidence based treatment. The heart failure nursing service is invaluable in bridging the gap between primary and secondary care. Local guidelines, together with continuing education of GPs and practice nurses, and the new General Medical Services contract, should further increase the uptake of evidence based treatments at target doses. PMID- 15831611 TI - The role of nurses in the management of heart failure. AB - Care provided by specialist nurses has been shown to improve outcomes for patients with chronic heart failure (CHF), significantly reducing the number of unplanned readmissions, length of hospital stay, hospital costs, and mortality. Most patients develop CHF as a result of coronary artery disease. Once cardiac damage has occurred, the risk of developing heart failure can be reduced by providing appropriate treatment at appropriate dosages. While cardiac rehabilitation clinics provide an opportunity to check drug usage, their prime focus is on optimising patients' physical well being following a heart attack. In addition, evidence suggests that general practitioners are frequently reluctant to initiate appropriate treatments and to up-titrate drug dosages even for patients with diagnosed heart failure. Therefore, to ensure that these patients are not left on starting doses of medications many hospitals are now setting up nurse led post-myocardial infarction (MI) clinics. The Omada programme is a secondary care based, nurse led model of care set up in 1999 to improve the management of CHF by providing appropriate patient education within a nurse led clinic setting, optimising evidence based medication and fostering partnership between health professionals in both primary and secondary care. The model of care is highly applicable to the post-MI setting, where it can ensure that patients receive better care at an earlier stage. PMID- 15831613 TI - Epidemiology and management of heart failure and left ventricular systolic dysfunction in the aftermath of a myocardial infarction. AB - Robust epidemiological data on the incidence of myocardial infarction (MI) are hard to find, but synthesis of data from a number of sources indicates that the average hospital in the UK should admit about two patients with a first MI and one recurrent MI per 1000 population per year. Possibly the most relevant data on the incidence, prevalence, and persistence of post-MI heart failure can be derived from the TRACE study. Most patients will develop heart failure or major left ventricular systolic dysfunction (LVSD) at some time after an MI, most commonly during the index admission. In up to 20% of cases this will be transient, but such patients still have a poor prognosis. There is likely to be around one patient discharged per thousand population per year with heart failure or major LVSD after an acute MI. It is important to organise care structures to ensure that patients with post-MI heart failure and LVSD are identified and managed appropriately. PMID- 15831614 TI - British Cardiac Society (BCS) Abstracts 2005. PMID- 15831617 TI - Gentamicin induced ototoxicity during treatment of enterococcal endocarditis: resolution with substitution by netilmicin. AB - Enterococcal endocarditis can be very difficult to eradicate, requiring prolonged treatment with a combination of a penicillin and an aminoglycoside. In this patient with a pacemaker associated enterococcal endocarditis, ototoxicity occurred due to total gentamicin dose despite plasma concentrations consistently within the treatment range. Substitution with netilmicin, without a break in aminoglycoside treatment, resulted in a rapid improvement in hearing and allowed the required course of aminoglycoside to be completed. The risk factors for ototoxicity with gentamicin are reviewed, in particular the dangers of increasing age and of multiple and prolonged courses. Close treatment monitoring does not totally avoid this risk, especially when prolonged aminoglycoside treatment is required. This case emphasises the need for prompt investigation and adequate, definitive treatment of enterococcal endocarditis to avoid the increased risk consequent on repeated courses of antibiotics. The resolution of the ototoxicity with netilmicin is consistent with other reports of lower cochleotoxicity than with other aminoglycosides. PMID- 15831618 TI - Lysyl oxidase deficiency: a new cause of human arterial dissection. AB - Spontaneous coronary artery dissection is a rare cause of myocardial ischaemia. The underlying mechanism is unknown but some dissections are associated with extracellular matrix disorganisation of genetic origin. A deficiency in extracellular matrix protein cross links has rarely been studied. A single clinical case of spontaneous coronary artery dissection is reported. Lysyl oxidase (LOX) and extracellular matrix organisation were investigated by skin immunohistology and polymerase chain reaction of LOX expression. Both approaches found a dramatic LOX decrease. LOX deficiency has a major role in human arterial wall organisation during development. The suspected mechanism is an elastin or collagen polymer cross linking deficiency. PMID- 15831619 TI - Acute myocardial infarction caused by a septic coronary embolism diagnosed and treated with a thrombectomy catheter. AB - Acute myocardial infarctions are common in bacteraemia but are seldom diagnosed during life. A 64 year old man with severe chest pain who had fever for several days due to possible bacteraemia was shown by ECG and echocardiography to have possible lateral infarction. Immediate coronary angiography showed possible thrombus in the left circumflex artery, which was treated by thrombectomy catheter. Bacterial thrombus was removed and was verified by histological examination. A stent was implanted without complications. Acute myocardial infarction caused by septic embolism is usually fatal; however, thrombectomy may be useful in these cases. PMID- 15831620 TI - Total relief of severe left ventricular outflow obstruction after spontaneous rupture of chordae tendineae in a patient with hypertrophic cardiomyopathy. AB - In hypertrophic cardiomyopathy (HCM), rupture of mitral chordae tendineae is infrequent and causes acute haemodynamic deterioration. A 38 year old male patient had chordae rupture leading to prolapse of both mitral leaflets and severe regurgitation, without change in symptomatic status. One year before, he had had mild mitral regurgitation and a resting left ventricle outflow tract of 105 mm Hg that disappeared in the present evaluation. In this unique case, worsening of mitral regurgitation was counterbalanced by total relief of the severe obstruction. This case report highlights the role of the mitral valve apparatus in the genesis of obstruction in HCM, further stimulating surgical techniques in which mitral repair can be the main procedure. PMID- 15831621 TI - Coronary air embolism treated with aspiration catheter. AB - Coronary air embolism remains a recognised complication of coronary catheterisation despite a strong emphasis on prevention. Current treatment consists of supportive measures with 100% oxygen and analgesia. Recent case reports describe the use of mechanical treatments aimed at dispersing or removing the air embolus with variable success. A case of coronary air embolism causing an acute coronary syndrome is described that was definitively treated with an aspiration system. The effectiveness of the aspiration system in the distal section of an obtuse marginal artery indicates that such dedicated aspiration systems may prove useful in the standard treatment of air embolism. PMID- 15831622 TI - Vasculitis masquerading as aortic valve endocarditis. AB - Small vessel vasculitis and endocarditis can both present with multisystem involvement and may present a diagnostic dilemma. Renal and cardiac involvement is common in small vessel vasculitis and rarely small vessel vasculitis may cause heart block. When a patient presents with diffuse symptoms, deteriorating renal function, and heart block, endocarditis and vasculitis should be included in the differential diagnosis. The case is discussed of a man with a history of aortic valve endocarditis who presented again with similar symptoms, deteriorating renal function, and heart block. There was no evidence of aortic valve endocarditis with abscess formation. A renal biopsy confirmed small vessel vasculitis and the patient responded promptly to immunosuppressive treatment. Correct diagnosis is essential in such cases, as immunosuppression in true endocarditis can be catastrophic. In this case, with the correct diagnosis, immunosuppression proved life saving and prevented erroneous aortic valve surgery. PMID- 15831623 TI - Novel use of Brocken brough needle in relieving membranous obstruction of the inferior vena cava. PMID- 15831624 TI - Electrocardiographic and echocardiographic features of a disconnected right ventricle. PMID- 15831625 TI - Right ventricular myocardial infarction. PMID- 15831626 TI - Traumatic ventricular septal defect and tricuspid regurgitation. PMID- 15831627 TI - Hypertrophic cardiomyopathy with mid ventricular obstruction, apical infarction and aneurysm formation. PMID- 15831628 TI - Mitral valve "kissing" vegetation. PMID- 15831629 TI - Ruptured sinus of Valsalva aneurysm. PMID- 15831630 TI - Late opening of the infarct related artery: an open or shut case? AB - Is there a place for the late opening of infarct related arteries, beyond the time window for myocardial salvage? PMID- 15831631 TI - Warfarin: a case history in pharmacogenetics. AB - Both hereditary and environmental factors contribute to inter-individual variability in drug response. The considerable interest in the role of genes has to be balanced with the contribution of external influences. Warfarin provides a useful case study of the need to integrate both genetic and non-genetic information when selecting the right dose for a patient. This article discusses the latest data on genotype and warfarin sensitivity and the efforts to incorporate this information into normograms. Exploring the genetics of warfarin response will lead not only to safer prescribing but elucidate the mechanism of action of warfarin and enable the development of new anticoagulant drugs. PMID- 15831632 TI - Value of the myocardial performance index in myocardial infarction. AB - The myocardial performance index represents an easy and reproducible parameter of both systolic and diastolic left ventricular function for the risk stratification of patients following acute myocardial infarction. PMID- 15831633 TI - Diagnosis of myocarditis by cardiovascular magnetic resonance. PMID- 15831634 TI - Traumatic aortic and mitral valve injury following blunt chest injury with a variable clinical course. AB - Blunt trauma is uncommonly followed by intracardiac valvar injuries. The resulting valvar insufficiency rapidly or progressively leads to congestive heart failure or death unless surgically corrected. Three patients with sustained blunt chest trauma were found to have two aortic valve and one mitral valve ruptures. They had variable clinical courses. However, after the diagnosis was established, surgical intervention was attempted promptly, which consisted of two aortic valve replacements and one mitral valvoplasty. Their postoperative courses were uneventful. Careful observation and repeated physical examination, aided by echocardiography, were required after the blunt chest trauma. PMID- 15831635 TI - Infective endocarditis in Europe: lessons from the Euro heart survey. AB - OBJECTIVES: To describe the characteristics, treatment, and outcomes of active infective endocarditis (IE) in Europe. DESIGN: Prospective survey of medical practices in Europe. SETTING: 92 centres from 25 countries. PATIENTS: The EHS (Euro heart survey) on valvar heart disease (VHD) enrolled 5001 adult patients between April and July 2001. Of those, 159 had active IE. RESULTS: 118 patients (74%) had native IE and 41 (26%) had prosthetic IE. Mean (SD) age was 57 (16) years. Blood cultures were obtained for 113 patients (71%) before antibiotic treatment was started. Surgery was performed in 52% of patients. Reasons for surgery were heart failure in 60%, persistent sepsis in 40%, vegetation size in 48%, or embolism in 18%. Surgery was for implantation of mechanical prosthesis in 63%, bioprosthesis in 21%, aortic homograft in 5%, and valve repair in 11%. In hospital mortality was 12.6%, being 10.4% in the medical group and 15.6% in the surgical group. Among the total population of 5001 patients, only 50% of those with native VHD had been educated on endocarditis prophylaxis and only 33% regularly attended dental follow up. Of patients with IE who had had a procedure at risk during the preceding year only 50% had received adequate prophylaxis. CONCLUSIONS: The EHS on VHD shows that patients with active IE have a high risk profile and often undergo surgery. However, there are deficiencies in obtaining blood cultures and applying prophylaxis. Mortality remains high, which is a justification for the improvement of patient management through education and the implementation of guidelines. PMID- 15831636 TI - Atherosclerosis-like lesions of the aortic valve are common in adults of all ages: a necropsy study. AB - OBJECTIVE: To determine the presence and size of atherosclerosis-like lesions in aortic valves of unselected adults of all ages. DESIGN: Necropsy study. SETTING: Departments of Forensic Medicine and Pathology at the University of Oulu and the Oulu University Hospital in Finland. SUBJECTS: 48 consecutive unselected adult subjects (15 subjects aged 20-40 years, 17 aged 41-60, and 16 aged >or= 61) undergoing necropsy. MAIN OUTCOME MEASURES: Detection of the presence of atherosclerosis-like lesions and mineralisation in aortic valves, and morphometrical measurement of the size of lesions. RESULTS: None of the necropsy subjects had aortic stenosis and only two subjects had macroscopic calcification of the aortic valve. Of 48 subjects, however, 45 had an atherosclerosis-like subendothelial thickening above the elastic lamina on the aortic side of at least one of the valve leaflets. Of 15 young subjects aged 20-40 years, eight had a lesion in the right coronary cusp of the aortic valve and 12 had a lesion in at least one of the three aortic valve leaflets. Of 17 middle aged subjects, 16 had an early lesion in the right coronary cusp and all had a lesion in at least one of the valve leaflets. In the oldest age group, all 16 subjects had a lesion in every valve leaflet. The mean lesion area in the three valve leaflets varied from 0.1-0.2 mm(2) in young subjects, 0.5-0.8 mm(2) in middle aged, and 1.3-2.3 mm(2) in elderly subjects (p < 0.001). Microscopic calcification in the right coronary cusp of the aortic valve was observed in 12 of 17 middle aged and 14 of 16 elderly subjects but only in one young subject. CONCLUSIONS: Atherosclerosis-like lesions in the aortic valve are prevalent in adults of all age groups, including young subjects aged 20-40 years, suggesting that the disease process leading to aortic stenosis is common, often beginning in early adulthood. PMID- 15831637 TI - Ventricular non-compaction cardiomyopathy: CMR and pathology findings. PMID- 15831638 TI - Myocardial performance index for assessment of left ventricular outcome in successfully recanalised anterior myocardial infarction. AB - OBJECTIVE: To investigate whether the myocardial performance index (MPI) can predict left ventricular functional outcome in patients with early recanalisation after anterior acute myocardial infarction (MI) and to determine when the index should be measured. DESIGN: MPI was measured serially by two dimensional Doppler echocardiography after successful percutaneous coronary intervention (PCI). Left ventricular function was evaluated by echocardiography and left ventriculography. To assess coronary microvascular damage, the coronary flow velocity pattern was measured immediately after PCI with a Doppler guidewire. SETTING: Hiroshima City Asa Hospital. PATIENTS: 32 consecutive patients with their first anterior acute MI who had complete occlusion of left anterior descending coronary artery. INTERVENTIONS: Successful PCI within six hours of symptom onset. MAIN OUTCOME MEASURES: Left ventricular anterior wall motion score index (A-WMSI), left ventricular end diastolic pressure (LVEDP), left ventricular ejection fraction (LVEF), and left ventricular end diastolic volume (LVEDV). RESULTS: There was a significant negative correlation between MPI on day 2 and the coronary diastolic deceleration time (r = -0.66, p < 0.002), as well as a significant positive correlation with the coronary diastolic deceleration rate (r = 0.74, p < 0.0001). MPI on day 2 was significantly correlated with the short and long term changes of A-WMSI and with the short term changes of LVEDP. Furthermore, MPI on day 2 was significantly correlated with the short and long term changes of LVEF (r = -0.52, p < 0.003, and r = -0.64, p < 0.0008, respectively) and of LVEDV (r = 0.51, p < 0.003, and r = 0.41, p < 0.05, respectively). CONCLUSIONS: Doppler derived MPI on day 2, representative of the early coronary microvascular state, can predict the left ventricular functional outcome after early successful recanalisation of a patient's first anterior acute MI. PMID- 15831639 TI - The "tricolore" sign of heart failure. PMID- 15831640 TI - Long term cardiovascular effects of oral antidiabetic agents in non-diabetic patients with insulin resistance: double blind, prospective, randomised study. AB - OBJECTIVE: To study the long term cardiovascular effects of oral antidiabetic agents in non-diabetic patients with insulin resistance. PATIENTS: 181 African American subjects with insulin resistance and normal glucose tolerance test were randomised to receive glipizide 5 mg/day (n = 25), metformin 500 mg/day (n = 59), or placebo (n = 97) for 24 months. Insulin sensitivity, glucose tolerance, lipid profile, left ventricular mass (echocardiography), aortic distensibility (echocardiography, blood pressure), aortic pulse wave velocity (PWV, carotid to femoral artery, Doppler) were measured at baseline and at 12 and 24 months after randomisation. RESULTS: A significant increase in PWV was observed in both glipizide (mean (SEM) change at 24 months 2.8 (2.7) m/s, p = 0.012) and metformin (2.2 (0.7) m/s, p = 0.01) groups during the follow up period. In contrast, PWV remained unchanged in the placebo group. The increase in PWV in the treatment groups was significant compared with placebo (analysis of variance p < 0.05). Other cardiovascular or metabolic variables did not change significantly compared with placebo during follow up. CONCLUSIONS: The observed increase in PWV is consistent with a decrease in the elastic properties of the aorta. The use of oral antidiabetic agents for the prevention of cardiovascular complications in non-diabetic African Americans with insulin resistance needs to be critically evaluated. PMID- 15831641 TI - Ethnic differences in the prevalence and aetiology of left ventricular systolic dysfunction in the community: the Harrow heart failure watch. AB - OBJECTIVE: To assess ethnic differences in the prevalence and aetiology of left ventricular systolic dysfunction (LVSD) in the community. DESIGN: Community cohort study. All patients underwent echocardiography and those found to have LVSD underwent myocardial perfusion imaging with or without coronary angiography to diagnose underlying coronary artery disease (CAD). SETTING: Seven representative general practices in Harrow, UK, a community hospital, and a local district general hospital. PATIENTS: 1392 patients >or= 45 years old randomly selected from the computer records of seven general practices. MAIN OUTCOME MEASURES: The prevalence and aetiology of LVSD in the community, assessing differences between white and non-white populations, and the proportion of patients with LVSD with undiagnosed CAD. RESULTS: 734 patients (53%) attended, 518 (71%) white and 216 (29%) non-white, the majority South Asian. Thirty nine patients (5.5%) had probable LVSD and 25 (3.5%) definite LVSD. No significant differences in prevalence were seen with ethnicity. CAD underlay most cases of LVSD. Non-white patients had a higher prevalence of CAD as the underlying aetiology of significant LVSD than white patients (100% v 56%, p = 0.04) and a trend towards less alcoholic cardiomyopathy. 8% of patients with LVSD had undiagnosed CAD. CONCLUSIONS: LVSD is common. White and non-white patients have a similar overall prevalence of LVSD. Non-white patients, the majority South Asians in this study, have a higher prevalence of CAD as the underlying cause for LVSD than white patients. CAD underlies most cases of LVSD in the community, although it may be undiagnosed unless formally assessed. PMID- 15831642 TI - Long term biventricular resynchronisation therapy in advanced heart failure: effect on neurohormones. AB - OBJECTIVE: To assess prospectively the effect of cardiac resynchronisation therapy (CRT) on New York Heart Association (NYHA) functional class, cardiac function, cardiopulmonary exercise performance, and neurohormonal activation during 24 months' follow up. DESIGN: Controlled study. PATIENTS AND RESULTS: 124 patients with severe congestive heart failure (ejection fraction < 35%, NYHA III IV) and left bundle branch block (QRS duration > 150 ms) were enrolled (control group, n = 59; CRT group, n = 65) and followed up at 1, 3, 12, and 24 months. Compared with the control group, CRT led to significant short and long term improvements in functional NYHA functional class (mean (SEM) 2.1 (0.4) v 2.8 (0.4) at 24 months, p < 0.05), mean ejection fraction (25.7 (4)% v 21.1 (5)% at 24 months, p < 0.05), peak Vo(2) (16.8 (3.9) v 12.6 (3.5) ml/kg x min at 24 months, p < 0.01), and Vo(2) at anaerobic threshold (14.4 (3.7) v 10.8 (3.2) ml/kg x min at 24 months, p < 0.05). In addition, CRT for one and 12 months significantly decreased the plasma concentrations of noradrenaline (norepinephrine) and N-terminal fragment of pro-brain natriuretic peptide, whereas no changes were observed for other neurohormones such as antidiuretic hormone, aldosterone, and endothelin. CONCLUSION: Long term CRT ( 0.6 cm provided a sensitivity of 80%, a specificity of 51%, and a negative predictive value of 80% for the prediction of viability in akinetic segments. DSE had an excellent specificity (92%) but a modest sensitivity (60%) in akinetic segments. A combination of improvement at DSE or DWT > 0.8 cm improved sensitivity (90% v 60%, p < 0.001) and negative predictive value (92% v 78%, p = 0.03) in akinetic segments compared with DSE alone. This was achieved with some loss in specificity (75% v 92%, p = 0.01) and positive predictive value (71% v 82%, p = 0.79). CONCLUSIONS: DWT measurement may improve the sensitivity of DSE for the detection of myocardial viability. Akinetic segments with DWT > 0.8 cm have a good chance of recovery despite the absence of contractile reserve during DSE. Further testing may be required before excluding myocardial viability in these cases. PMID- 15831645 TI - Effects of magnesium on atrial fibrillation after cardiac surgery: a meta analysis. AB - OBJECTIVES: To assess the efficacy of the administration of magnesium as a method for the prevention of postoperative atrial fibrillation (AF) and to evaluate its influence on hospital length of stay (LOS) and mortality. METHODS: Literature search and meta-analysis of the randomised control studies published since 1966. RESULTS: 20 randomised trials were identified, enrolling a total of 2490 patients. Study sample size varied between 20 and 400 patients. Magnesium administration decreased the proportion of patients developing postoperative AF from 28% in the control group to 18% in the treatment group (odds ratio 0.54, 95% confidence interval (CI) 0.38 to 0.75). Data on LOS were available from seven trials (1227 patients). Magnesium did not significantly affect LOS (weighted mean difference -0.07 days of stay, 95% CI -0.66 to 0.53). The overall mortality was low (0.7%) and was not affected by magnesium administration (odds ratio 1.22, 95% CI 0.39 to 3.77). CONCLUSION: Magnesium administration is an effective prophylactic measure for the prevention of postoperative AF. It does not significantly alter LOS or in-hospital mortality. PMID- 15831646 TI - Apical left ventricular aneurysm presenting with malignant ventricular tachycardia responsive to aneurysmectomy. PMID- 15831647 TI - Relation between plasma amino-terminal propeptide of procollagen type III and left ventricular longitudinal strain in essential hypertension. AB - OBJECTIVE: To investigate whether myocardial fibrosis assessed non-invasively is related to left ventricular (LV) longitudinal systolic function in patients with essential hypertension. DESIGN: The study consisted of 30 control subjects and 40 patients with hypertension with normal LV ejection fraction. Tissue Doppler echocardiography was performed to assess LV longitudinal systolic strain from the apical views. Mean strain was calculated from the basal and mid segments. Plasma concentrations of the amino-terminal propeptide of type III procollagen (PIIINP) were measured. RESULTS: In the hypertension group, mean strain was significantly reduced (mean (SD) 13 (6)% v 21 (6)%, p < 0.01) and plasma PIIINP were increased compared with controls (3.0 (0.7) microg/l v 2.1 (0.3) microg/l, p < 0.001). A significant correlation was found between mean strain and PIIINP (r = -0.56, p < 0.001). In patients with abnormal diastolic filling (n = 21) mean strain was reduced compared with patients with normal LV filling (n = 19) (10 (6)% v 15 (6)%, p < 0.01) and the serological marker PIIINP was increased (3.5 (0.6) microg/l v 2.5 (0.5) microg/l, p < 0.001). CONCLUSIONS: There is a significant association between the extent of myocardial fibrosis and reduced LV longitudinal contractility. PMID- 15831648 TI - Scintigraphic diagnosis of acute pulmonary oedema. PMID- 15831649 TI - Evidence for inequalities in the management of coronary heart disease in Scotland. AB - OBJECTIVES: To investigate whether sex, age, and deprivation inequalities existed in the prescription of secondary preventive treatment for coronary heart disease (CHD) in Scottish general practice and whether these differences altered over time. DESIGN: 6 year cross sectional study based on general practice morbidity and prescribing data. SETTING: 55 primary care practices in Scotland. SUBJECTS: 14,435 patients with diagnosed CHD. MAIN OUTCOME MEASURE: Prescription of various groups of secondary preventive treatment in six study years. RESULTS: The use of all secondary prevention treatments increased over time (63.6% of patients with CHD in 1997 to 87.6% in 2002). After adjustments for age, sex, deprivation, co morbidities, and practice where appropriate, women received fewer secondary prevention treatments than men, a difference that increased over time (March 1997: adjusted odds ratio (OR) 0.9, 95% confidence interval (CI) 0.8 to 1.0; March 2002: OR 0.6, 95% CI 0.6 to 0.7). Sex differences were observed within each group of treatments studied. The oldest group of patients was less likely than the youngest group to receive any secondary preventive treatment in the year up to March 1997 (OR 0.6, 95% CI 0.5 to 0.7) but were more likely by 2002 (OR 1.3, 95% CI 1.1 to 1.5) to receive secondary prevention. The most affluent patients with CHD were significantly less likely to receive a statin between March 1998 and 2001 (March 1998 OR 0.6, 95% CI 0.5 to 0.9), a finding that disappeared by 2002 (OR 0.9, 95% CI 0.7 to 1.1). CONCLUSION: The results suggest that inequalities exist in the secondary prevention of CHD in Scotland. PMID- 15831650 TI - Inequalities in coronary revascularisation during the 1990s: evidence from the British regional heart study. AB - OBJECTIVE: To investigate the influence of age and social circumstances on probability of revascularisation among British men. DESIGN: Prospective population based study SETTING: 24 medium sized British towns, none of which contained a hospital undertaking coronary artery bypass surgery. SUBJECTS: 5814 surviving participants of the BRHS (British regional heart study), aged 52-73 years, with no history of revascularisation when responding to a questionnaire in November 1992. MAIN OUTCOMES: Incident coronary revascularisations, as documented in general practitioner records, over the following 7.1 years and coronary angiography investigations reported by men in a further questionnaire in November 1996. RESULTS: 160 men underwent at least one revascularisation during this period (4.2/1000 person-years). In multifactorial analysis, which included adjustment for incidence of major coronary heart disease or angina, a lower incidence of revascularisation was found among men aged over 65 years in November 1992 (hazard ratio 0.62, 95% confidence interval (CI) 0.44 to 0.87), among men with manual occupations (0.73, 95% CI 0.53 to 1.02), among men living in households possessing no car (0.44, 95% CI 0.24 to 0.80) or one car (0.60, 95% CI 0.42 to 0.87) compared with two or more cars, among council tenants (0.49, 95% CI 0.25 to 0.97), and among men living outside southern England (0.71, 95% CI 0.51 to 0.99). Only car ownership was related to the incidence of diagnostic angiography: the odds ratio for angiography for those owning fewer than two cars was 0.62 (95% CI 0.42 to 0.89). CONCLUSION: During the 1990s, there were major inequalities in the probability of undergoing coronary revascularisation between British men according to socioeconomic status, age, and geographic location. PMID- 15831651 TI - Massive thoracic aortic aneurysm. PMID- 15831652 TI - Is routine stenting for acute myocardial infarction superior to balloon angioplasty? A randomised comparison in a large cohort of unselected patients. AB - OBJECTIVE: To evaluate the impact of routine stenting, compared with balloon angioplasty, in unselected patients presenting with ST segment elevation myocardial infarction (STEMI). DESIGN: Randomised trial. SETTING: Tertiary referral centre. PARTICIPANTS: All patients presenting with STEMI randomly assigned to stenting or balloon angioplasty. No exclusion criteria were applied. MAIN OUTCOME MEASURE: The primary end point was combined death or reinfarction at one year's follow up. RESULTS: 1683 consecutive patients with STEMI were randomly assigned before angiography to stenting (n = 849) or balloon angioplasty (n = 834). A total of 785 patients (92.5%) in the stent group and 763 patients (91.5%) in the balloon group actually underwent primary angioplasty. The groups were comparable in terms of postprocedural TIMI (thrombolysis in myocardial infarction) flow, myocardial blush grade, and distal embolisation. No difference was observed in clinical outcome at both intention to treat (14% v 12.5%, not significant) and actual treatment analyses (12.4% v 11.3%, not significant). CONCLUSIONS: Compared with balloon angioplasty, routine stenting does not seem to reduce death and reinfarction in a large cohort of unselected patients with STEMI. PMID- 15831654 TI - Brugada syndrome precipitated by a tricyclic antidepressant. PMID- 15831653 TI - Late reopening of an occluded infarct related artery improves left ventricular function and long term clinical outcome. AB - OBJECTIVE: To assess effects on left ventricular (LV) function and on long term clinical outcome of late percutaneous transluminal coronary angioplasty (PTCA) of a chronically occluded infarct related artery. METHODS: 65 patients who underwent PTCA a mean (SD) of 6.0 (1.2) months after a previous myocardial infarction were divided in two groups according to dilated artery patency status after PTCA: group 1 (35 patients with TIMI (thrombolysis in myocardial infarction) grade 3 flow) and group 2 (30 patients with TIMI grade 0-2 flow). Echocardiography was performed at admission and at six months' follow up. A three year follow up was conducted with major adverse cardiac events (MACE) as end points. RESULTS: At follow up, group 1 had improved global LV ejection fraction (48.7% v 43.6%, p < 0.001) and LV indexed end diastolic and end systolic volumes (75 v 86 ml/m(2) and 40 v 53 ml/m(2), respectively, p = 0.011) compared with group 2. Kaplan-Meier analysis showed a higher incidence of cardiac death (p = 0.02) and MACE (p < 0.0001) in group 2. TIMI 3 after PTCA was an independent predictor of event-free survival at follow up. CONCLUSION: Late PTCA of a chronically occluded infarct related artery improves LV function, reduces cardiac death, and improves long term clinical outcome. PMID- 15831655 TI - Late outcome of Senning and Mustard procedures for correction of transposition of the great arteries. AB - OBJECTIVES: To evaluate late mortality and morbidity after an atrial switch procedure for correction of transposition of the great arteries (TGA) and to assess predictive factors for adverse outcome. SETTING: Tertiary referral centre. DESIGN AND PATIENTS: Retrospective follow up study of 137 patients surviving hospitalisation for TGA atrial switch procedure (Mustard or Senning) in a single institution and divided into two groups (simple and complex) depending on presurgical anatomy. Several surgical and follow up factors were evaluated during 16.7 (5.6) years' follow up. RESULTS: Late mortality was 5.1% (95% confidence interval 1.37% to 8.84%) with sudden death as the most common cause. No significant difference was found between Mustard and Senning procedures and between the complex and simple groups in terms of mortality. Independent predictive factors for late mortality were a history of supraventricular tachyarrhythmias and advanced New York Heart Association (NYHA) functional class during follow up. A very common finding was development of sinus node dysfunction (47.6%), which had no influence on mortality. There was little need for reintervention (5.1%) and relatively few cases of right ventricular systolic dysfunction (14.6%). During follow up, most patients (96.2%) were in NYHA functional class I-II. CONCLUSIONS: Overall long term outcomes of patients with atrial repair of TGA in the present era are encouraging in terms of late mortality and quality of life. Nevertheless, better outcomes may be offered through improved diagnostic methods for right ventricular function and better management of supraventricular tachyarrhythmias. PMID- 15831656 TI - Magnetic resonance imaging of anomalous origin of the left coronary artery from the pulmonary artery (Bland-White-Garland syndrome). PMID- 15831657 TI - Evolution of the management approach for pulmonary atresia with intact ventricular septum. AB - OBJECTIVE: To review the evolution of the management approach for pulmonary atresia with intact ventricular septum (PAIVS) in the past two decades and to assess its impact on patient outcomes. DESIGN AND PATIENTS: Retrospective review of the management and outcomes of 94 patients (55 male patients) with PAIVS diagnosed between July 1980 and August 2003. SETTINGS: Tertiary paediatric cardiac centre. RESULTS: Seven patients died before interventions. Of the remaining 87 patients who underwent intervention at a median age of 9 days (from 1 day to 2 years), 12 had right ventricular outflow tract reconstruction (RVOTR), 42 had closed pulmonary valvotomy (CPV), and 15 had laser assisted valvotomy with balloon valvoplasty. A systemic-pulmonary shunt was inserted in 18 patients, six of whom had subsequent RVOTR (n = 4) or laser assisted valvotomy (n = 2). Since 1990, catheter intervention accounted for 38% (17 of 45) of the right ventricular outflow procedures. The mean (SEM) freedom from reintervention was 93 (7)%, 71 (12)%, and 57 (13)% after RVOTR, 75 (7)%, 40 (8)%, and 14 (6)% after CPV, and 54 (13)%, 24 (12)%, and 16 (10)% after laser assisted valvotomy at one month, six months, and one year, respectively (RVOTR versus CPV, p < 0.001; RVOTR versus laser assisted valvotomy, p = 0.001). Low cardiac output syndrome was significantly less common after catheter intervention than after RVOTR (0% v 44%, p = 0.003) or CPV (0% v 29%, p = 0.01). The overall mean (SEM) survival was 77 (5)% and 70 (5)% at one and five years, respectively, and the overall mortality was 33% (29 of 87). There were no significant differences in survival between the three groups. CONCLUSIONS: Multiple interventions are often required in the treatment algorithm of PAIVS. The shift towards increased use of the transcatheter approach has reduced the occurrence of postprocedural low cardiac output syndrome. PMID- 15831658 TI - Double mycotic aneurysm of the descending thoracic aorta. PMID- 15831659 TI - Inhibition of angiotensin II induced endothelin-1 gene expression by 17-beta oestradiol in rat cardiac fibroblasts. AB - OBJECTIVE: To examine whether 17-beta-oestradiol (E(2)) may alter angiotensin II (Ang II) induced cell proliferation and to identify the putative underlying signalling pathways in rat cardiac fibroblasts. DESIGN: Cultured rat cardiac fibroblasts were preincubated with E(2) then stimulated with Ang II. [(3)H]Thymidine incorporation and endothelin-1 (ET-1) gene expression were examined. The effect of E(2) on Ang II induced NADPH oxidase activity, reactive oxygen species (ROS) formation, and extracellular signal regulated kinase (ERK) phosphorylation were tested to elucidate the intracellular mechanism of E(2) in proliferation and ET-1 gene expression. RESULTS: Ang II increased DNA synthesis, which was inhibited with E(2) (1-100 nmol/l). E(2), but not 17-alpha-oestradiol, inhibited Ang II induced ET-1 gene expression as shown by northern blotting and promoter activity assay. This effect was prevented by co-incubation with the oestrogen receptor antagonist ICI 182,780 (1 micromol/l). E(2) also inhibited Ang II increased NADPH oxidase activity, ROS formation, ERK phosphorylation, and activator protein-1 mediated reporter activity. CONCLUSIONS: The results suggest that E(2) inhibits Ang II induced cell proliferation and ET-1 gene expression, partially by interfering with the ERK pathway through attenuation of ROS generation. Thus, this study provides important new insight regarding the molecular pathways that may contribute to the proposed beneficial effects of oestrogen on the cardiovascular system. PMID- 15831660 TI - Irbesartan significantly reduces C reactive protein concentrations after 1 month of treatment in unstable angina. PMID- 15831661 TI - Subclinical pulmonary vein narrowing after ablation for atrial fibrillation. PMID- 15831662 TI - Prediction and prevention of sudden cardiac death in heart failure. PMID- 15831663 TI - Assessment of diastolic function: what the general cardiologist needs to know. PMID- 15831664 TI - Stem cell therapy for myocardial repair. PMID- 15831665 TI - Diarrhea associated with Cryptosporidium parvum among young children of the Nile River Delta in Egypt. AB - Over a 2-year period, the prevalence and clinical characteristics of Cryptosporidium-associated diarrhea in the Nile River Delta of Egypt was studied. A stool sample was obtained from children with diarrhea attending one of two study hospitals and of the 1275 children evaluated, 214 (17%) were found to be infected with Cryptosporidium. Younger age was a risk factor for developing Cryptosporidium-associated diarrhea. Children <12 months of age were 2.4 times more likely to be infected with Cryptosporidium (p<0.01) and children 12 to 23 months were 1.9 (p<0.05) times more likely to be infected with the organism as compared to older children. Breastfeeding had a trend towards protection against Cryptosporidium-associated diarrhea (p=0.07). Clinical findings associated with Cryptosporidium diarrhea included vomiting, persistent diarrhea and the need for hospitalization. Our data suggest that Cryptosporidium is common in Egyptian children and may be associated with severe diarrhea. PMID- 15831666 TI - Pretreatment blood concentrations of chloroquine in patients with malaria infection: relation to response to treatment. AB - Resistance of Plasmodium falciparum to chloroquine has been reported in many areas in Ghana. Most of these reports, which are from hospital-based studies, indicate RI and RII rather than RIII type of resistance. Since high pretreatment levels of chloroquine have also been measured in patients with malaria infection in Ghana, we hypothesized that the 'added effect' of the pretreatment ingested drug to the full dose given at the hospital may be responsible for the low proportion of RIII type of resistance observed. To ascertain this, pretreatment blood levels of chloroquine were correlated with treatment outcomes in 231 paediatric malaria patients, referred to a major hospital in Ghana. The rate of parasite clearance and prevalence of recrudescence, 14 days post-treatment, were determined for each patient. Results from this study showed no correlation between pretreatment chloroquine levels and day 0 parasitaemia. Two hundred and seven patients (89.6 per cent) had parasites that were sensitive to chloroquine whilst 24 (10.4 per cent) had resistant parasites. Of the latter group 17, six, and one patients had P. falciparum parasites, which were resistant at RI, RII and RIII levels, respectively. Seventy-five per cent of the patients without any detectable pretreatment blood chloroquine had parasites that were sensitive to chloroquine whilst 89.8 per cent, 98 per cent, and 100 per cent with pretreatment blood chloroquine concentration ranges of 0.5--100.5 ng/ml, 100.5--200 ng/ml, and >200 ng/ml, respectively, had chloroquine-sensitive parasites. An inverse relationship was thus observed between pretreatment blood chloroquine concentration and the degree of resistance in this study. We conclude that pre hospital treatment ingested chloroquine contributes significantly to the resolution of malaria in children in Ghana, in the presence of chloroquine resistance. PMID- 15831667 TI - Late presentation of tracheobronchial foreign body aspiration in children. AB - The diagnosis of tracheobronchial foreign body (FB) aspiration in children is often delayed or overlooked. The aim of this study is to examine the causes of delay (>1 month) in the diagnosis and to study the complications of late presentation and their management. During the period between July 1993 and August 2002, 128 patients with suspected FB aspiration were admitted to KKUH, Riyadh, Saudi Arabia. Twenty-eight patients out of 128 patients presented late (>1 month). The medical records of those 28 patients were reviewed in respect to: history of FB aspiration, physical signs of symptoms, reason for delayed presentation, radiological investigation, complications and their management, and follow-up. There were 16 males and 12 females; the average age was 3.28 years (range 1--11 years). All of them experienced chronic cough at presentation. Thirteen (48 per cent) children had a history suggestive of FB aspiration. Seventeen (63 per cent) children presented with complications that included pneumonia (n=13), bronchiectasis (n=3), and bronchoesophageal fistula (n=1). The diagnostic delay was attributed to physician misdiagnosis (n=9), failure by parents to seek early medical advice (n=4), patients left against medical advice (n=1), and the cause of delay was unknown in the remaining 14 children. Rigid bronchoscopy was performed in all patients. FB was found in all cases except four. Complications were treated as follows: pneumonia with intravenous antibiotics, bronchiectasis conservatively and bronchoesophageal fistula repaired surgically. Follow-up ranged from 6 to 48 months. Diagnostic delay of tracheobronchial FB aspiration has significant morbidity in children. A high index of suspicion and early referral are essential in preventing such complications. PMID- 15831668 TI - Varicella zoster seroprevalence in children less than 5 years old. AB - This study was designed to evaluate the age-specific varicella-zoster virus (VZV) seroprevalence in children less than 5 years old who presented at a healthy child outpatient clinic and to compare the results with the data from other countries. The study was a cross-sectional study determining the prevalence of serum IgG against VZV in children who presented to the Healthy Child Outpatient Clinic of the Gazi University Medical Faculty and who were aged between 9 months and 5 years, in the 3rd--97th percentile as regards height and weight, not suffering from any disease, and without a history of vaccination against varicella. The information on the children was obtained from a questionnaire, by physical examination, and from patient files. Serum samples were obtained from babies and children at 9, 15, 24, 36, 48, and 60 months. The 295 serum samples were kept at -20 degrees C following centrifugation until used for serologic analysis (ELISA). The 292 children of the study group consisted of 168 males (57.5 per cent) and 124 females (42.5 per cent). VZV antibodies were found to be positive in 65 children aged between 9 months and 5 years (22.3 per cent); 22.0 per cent in males and 22.6 per cent in females with no statistically significant difference between the sexes (p>0.05). The VZV seroprevalence was highest at the 48th and 60th months and this difference was statistically significant (p=0.000). PMID- 15831669 TI - Molecular analysis of Iranian families with sickle cell disease. AB - Sickle hemoglobin is a mutant hemoglobin in which valine has been substituted for the glutamic acid normally at the sixth amino acid of the beta-globin chain. Detection of the single base pair mutation at codon 6 of the beta-globin gene is important for the prenatal diagnosis of sickle cell anemia and sickle cell disease. Application of the polymerase chain reaction technology to detect sickle cell patients and heterozygous carriers in a group of patients suspected for sickle cell disease was carried out. The sample was composed of 52 normal individuals and 52 unrelated outpatients who were attending the Hematology Research Center. All patients were interviewed. Results of their medical histories, physical examination, and the hematological analysis were recorded. The blood samples were collected in EDTA and genomic DNA was extracted from leukocytes. An amplified 110 base pair fragment of the beta-globin gene containing codon 6, was digested with the restriction enzyme MS II, and electerophoresed in 3 per cent agarose. We have established the technical condition for detection of sickle cell disease using a PCR assay. Fifteen patients having haemoglobin (Hb SS) and 37 patients in the heterozygous state (Hb AS) were identified. We confirm that the normal controls have the Hb AA genotype. The standardization of a highly sensitive and specific diagnostic test for sickle cell disease permitted us to identify the normal controls, the homozygotes and heterozygotes. This amplification method is rapid, sensitive and simple, and also has application research that is important for the prenatal diagnosis of sickle cell disease. PMID- 15831670 TI - Concurrent outbreak of leptospirosis and dengue in Mumbai, India, 2002. AB - This prospective study was undertaken to investigate the possibility of a concurrent outbreak of leptospirosis and dengue and to describe the clinical illnesses. From 20 June to 14 November 2002, children who presented to our hospital with a suspected diagnosis of leptospirosis or dengue were admitted. In every child with suspected leptospirosis, a screening latex agglutination test was carried out to detect anti-Leptospira antibodies. The diagnosis of leptospirosis was confirmed by a positive enzyme-linked immunosorbent assay (ELISA) test or microagglutination test. The diagnosis of dengue was confirmed by a positive IgM antibody capture ELISA test. Clinical features in the leptospirosis and leptospirosis-negative groups, and dengue and dengue-negative groups were analysed. Of 90 children screened, 15 (16.7 per cent) had leptospirosis. Two children with Weil's disease died and the remaining 13 responded well to intravenous penicillin. Five clinical features were significantly associated with leptospirosis, namely conjunctival suffusion (p=0.007), haemorrhage (p=0.020), abdominal pain (p=0.011), hepatosplenomegaly (p=0.044), and oedema (p=0.007). As the number of these five features concomitantly present increased, the chances of the child having leptospirosis also increased significantly (p<0.0001). Of 90 children screened, 16 (17.8 per cent) had dengue. All responded well to the treatment and went home. Two clinical features were significantly associated with dengue, namely arthralgia (p=0.020) and thrombocytopenia (p=0.001). If both these features were present, the chances of the child having dengue increased significantly (p=0.001). Our study shows that a concurrent outbreak of leptospirosis and dengue had occurred in the slums of Mumbai city. PMID- 15831671 TI - Sudden unexpected child deaths: forensic autopsy results in cases of sudden deaths during a 5-year period. AB - The aim of the present study was to determine the incidence of various causes of sudden unexpected child deaths (SUCD) and to assess the importance of an autopsy in predicting the likelihood of finding a cause of death. A retrospective analysis of autopsy findings in 97 cases of SUCD between the ages of 0--11 years was undertaken at the Council of Forensic Medicine, Ankara during a 5-year period (1995--2000). Cases were classified as explained causes (80.42 per cent) and sudden infant death syndrome (SIDS) (19.58 per cent). A total of 25.77 per cent of the deaths occurred in the neonatal period, 45.31 per cent of them in the first year of life and the remaining 28.86 per cent after 1 year of life. The causes of neonatal deaths were respiratory pathology (five cases), birth complications (four cases), gastrointestinal pathology (one case), homicide (10 cases), and SIDS (five cases). The incidence of SIDS in the newborn period was 33 per cent. The incidence of unexplained causes of deaths in the postneonatal period was 31 per cent and the causes of deaths were respiratory pathology (15 cases), aspiration (five cases), gastrointestinal pathology (four cases), SIDS (14 cases), and other causes (four cases). The study of an entire population provides more reliable data regarding causes of sudden unexpected child deaths than does the study of small groups and it is also recommended that in addition to a through evaluation, a detailed autopsy must be performed for each case in experienced centers. PMID- 15831672 TI - CrkI and CrkII function as key signaling integrators for migration and invasion of cancer cells. AB - Crk adaptor proteins play an important role during cellular signaling by mediating the formation of protein complexes. Increased levels of Crk proteins are observed in several human cancers and overexpression of Crk in epithelial cell cultures promotes enhanced cell dispersal and invasion, implicating Crk as a regulator of invasive responses. To determine the requirement of Crk for invasive signals, we targeted the CRKI/II gene by RNA interference. Consistent knockdown of CrkI/II was observed with two small interfering RNA targeting sequences in all human cancer cell lines tested. CrkI/II knockdown resulted in a significant decrease in migration and invasion of multiple malignant breast and other human cancer cell lines (MDA-231, MDA-435s, H1299, KB, and HeLa). Moreover, CrkI/II knockdown decreased cell spreading on extracellular matrix and led to a decrease in actin stress fibers and the formation of mature focal adhesions. Using immunohistochemistry, we show elevated CrkI/II protein levels in patients with breast adenocarcinoma. Together, these studies identify Crk adaptor proteins as critical integrators of upstream signals for cell invasion and migration in human cancer cell lines and support a role for Crk in metastatic spread. PMID- 15831673 TI - Extracellular matrix metalloprotease inducer-expressing head and neck squamous cell carcinoma cells promote fibroblast-mediated type I collagen degradation in vitro. AB - Until recently, tumor progression has been considered a multistep process defined by tumor cell mutations and the importance of the surrounding stroma poorly understood. It is now recognized that matrix-degrading enzymes that promote tumor cell invasion are elaborated by both tumor cells and fibroblasts in vivo. To determine the relative role of tumor cell-derived proteases compared with fibroblast-derived proteases, coculture experiments were done with each cell type using an in vitro model of type I collagen degradation. Head and neck squamous cell carcinoma cells in coculture with normal dermal fibroblasts showed matrix degradation, but neither cell type alone produced this effect. Manipulating the in vitro coculture environment showed that collagenolysis in this model was a result of fibroblast-derived matrix metalloproteases (MMP). To explore the possible role of extracellular matrix metalloprotease inducer (EMMPRIN) in this interaction, transfection of EMMPRIN into a cell line with low endogenous EMMPRIN expression was done and showed a significant increase in collagenolysis. Inhibition of collagenolysis with a tissue inhibitor of metalloprotease-2 (TIMP 2) and a synthetic furin inhibitor was observed but not with TIMP-1, which suggested a possible role for membrane type-1 MMP. These results suggest that fibroblast-derived MMPs but not those from tumor cells are important for in vitro collagenolysis and that this process is promoted by tumor cell-expressed EMMPRIN. PMID- 15831674 TI - Letrozole-, anastrozole-, and tamoxifen-responsive genes in MCF-7aro cells: a microarray approach. AB - Antiestrogens and aromatase inhibitors are important drugs in the treatment of estrogen-dependent breast cancer. To investigate the effects of these drugs on gene expression in breast cancer cells, we treated estrogen receptor-positive MCF 7 cells stably transfected with the aromatase gene (known as MCF-7aro cells) with testosterone, 17 beta-estradiol, two aromatase inhibitors (letrozole and anastrozole), and an antiestrogen (tamoxifen). We found that testosterone or 17 beta-estradiol induced the proliferation of MCF-7aro cells at a rate six times faster than the untreated cells. In addition, the testosterone-induced proliferation of MCF-7aro cells was effectively suppressed by letrozole, anastrozole, or tamoxifen. Microarray analyses on Affymetrix Human Genome U133A GeneChips (Affymetrix, Santa Clara, CA) were carried out using total RNA isolated from the control and treated cells. At the false discovery rate of 0.05 and a minimum fold-change criteria of 1.5, 104 genes were identified that were up regulated and 109 genes were identified that were down-regulated by both androgen and estrogen. More than 50% of these hormone-regulated genes were counter regulated by all three inhibitors and >90% were counter-regulated by at least one of the inhibitors. Comparing the effect of each inhibitor on gene expression, we observed that letrozole and anastrozole are more similar in terms of the genes they affect compared with treatment with tamoxifen. To validate the gene expression profiles identified from microarray analyses, the expression patterns of 13 representative genes were examined by Northern analysis. Finally, the genes identified as statistically significant were classified based on their expression patterns and biological function/pathways. The results of this study provide us with a better understanding of the actions of both aromatase inhibitors and antiestrogens at the molecular level. We believe that the results of this study serve as the first step in identifying unique expression patterns following drug treatment, and that this will ultimately be useful in customizing patient treatment strategies for hormone-dependent breast cancer. PMID- 15831675 TI - E1A sensitizes cancer cells to TRAIL-induced apoptosis through enhancement of caspase activation. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to induce apoptosis of cancer cells. Sensitization of cancer cells to TRAIL, particularly TRAIL-resistant cancer cells, could improve the effectiveness of TRAIL as an anticancer agent. The adenovirus type 5 E1A that associates with anticancer activities including sensitization to apoptosis induced by tumor necrosis factor is currently being tested in clinical trials. In this study, we investigated the sensitivity to TRAIL in the E1A transfectants ip1-E1A2 and 231 E1A cells and the parental TRAIL-resistant human ovarian cancer SKOV3.ip1 and TRAIL-sensitive human breast cancer MDA-MB-231 cells. The results indicated that the percentage of TRAIL-induced apoptotic cells was significantly higher in the E1A transfectants of both cell lines than it was in the parental cell lines. To further investigate the cellular mechanism of this effect, we found that E1A enhances TRAIL-induced activation of caspase-8, caspase-9, and caspase-3. Inhibition of caspase-3 activity by a specific inhibitor, Z-DEVD-fmk, abolished TRAIL-induced apoptosis. In addition, E1A enhanced TRAIL expression in ip1-E1A2 cells, but not in 231-E1A cells, and the anti-TRAIL neutralizing antibody N2B2 blocked the E1A-mediated bystander effect in vitro. Taken together, these results suggest that E1A sensitizes both TRAIL-sensitive and TRAIL-resistant cancer cells to TRAIL-induced apoptosis, which occurs through the enhancement of caspase activation; activation of caspase-3 is required for TRAIL-induced apoptosis; and E1A-induced TRAIL expression is involved in the E1A-mediated bystander effect. Combination of E1A and TRAIL could be an effective treatment for cancer. PMID- 15831676 TI - Akt activation, but not extracellular signal-regulated kinase activation, is required for SDF-1alpha/CXCR4-mediated migration of epitheloid carcinoma cells. AB - Emerging evidence shows that the stromal cell-derived factor 1 (SDF-1)/CXCR4 interaction regulates multiple cell signaling pathways and a variety of cellular functions such as cell migration, proliferation, and survival. There is little information linking the cellular functions and individual signaling pathways mediated by SDF-1 and CXCR4 in human cancer cells. In this study, we have shown that human epitheloid carcinoma HeLa cells express functional CXCR4 by reverse transcription-PCR, immunofluorescent staining, and 125I-SDF-1alpha ligand binding analyses. The treatment of HeLa cells with recombinant SDF-1alpha results in time dependent Akt and extracellular signal-regulated kinase 1/2 (ERK1/2) activations. The SDF-1alpha-induced Akt and ERK1/2 activations are CXCR4 dependent as confirmed by their total inhibition by T134, a CXCR4-specific peptide antagonist. Cell signaling analysis with pathway-specific inhibitors reveals that SDF-1alpha induced Akt activation is not required for ERK1/2 activation and vice versa, indicating that activations of Akt and ERK1/2 occur independently. Functional analysis shows that SDF-1alpha induces a CXCR4-dependent migration of HeLa cells. The migration can be totally blocked by phosphoinositide 3-kinase inhibitors, wortmannin or LY294002, whereas mitogen-activated protein/ERK kinase inhibitors, PD98059 and U0126, have no significant effect on SDF-1alpha-induced migration, suggesting that Akt activation, but not ERK1/2 activation, is required for SDF 1alpha-induced migration of epitheloid carcinoma cells. PMID- 15831677 TI - Fos-related antigen 1 modulates malignant features of glioma cells. AB - Malignant gliomas, and high-grade gliomas (HGG) in particular, are nonmetastasizing but locally infiltrating, hypervascularized brain tumors of poor prognosis. We found previously that a c-fos-inducible vascular endothelial growth factor D is ubiquitously up-regulated in HGG grade IV, glioblastoma multiforme, and that glioblastoma multiforme overexpress Fos-related antigen 1 (Fra-1) rather than the c-Fos. We have thus become interested in the role Fra-1 may play in malignant glioma progression/maintenance, because Fra-1 has the capacity to modulate transcription of a variety of target genes. In this work, we have analyzed the biological effects of ectopic Fra-1 expression or Fra-1 knockdown in malignant glioma cells. Ectopic Fra-1 induced prominent phenotypic changes in all three malignant glioma cell lines examined: H4, U-87 MG, and A-172 MG. These changes were reflected in cells becoming more elongated with larger number of cellular processes. Furthermore, Fra-1 transgene caused H4 cells, which do not form tumor xenografts, to regain tumorigenic capacity. The genotype of these cells changed too, because 50 of 1,056 genes examined became either up-regulated or down-regulated. Conversely, Fra-1 knockdown altered prominently the morphology, anchorage-independent growth, tumorigenic potential, and Fra-1 effector expression, such as vascular endothelial growth factor D, in HGG cells. For example, cells transfected with antisense fra-1 showed shorter cellular processes than the control cells that did not grow in agar, and their tumorigenic potential was significantly diminished. Thus, Fra-1 may likely play an important role in the maintenance/progression of malignant gliomas and potentially represents a new target for therapeutic interventions. PMID- 15831678 TI - Just one question: If one question works, why ask several? PMID- 15831679 TI - Efficacy, effectiveness, and the evaluation of public health interventions. PMID- 15831680 TI - Suicide risk after spousal suicide or psychiatric admission: effects of assortative mating on heritable traits compared with environmental explanations. PMID- 15831681 TI - Embodiment: a conceptual glossary for epidemiology. AB - This construct and process are central to ecosocial theory and epidemiological inquiry. Recognising that we, as humans, are simultaneously social beings and biological organisms, the notion of "embodiment" advances three critical claims: (1) bodies tell stories about-and cannot be studied divorced from--the conditions of our existence; (2) bodies tell stories that often--but not always--match people's stated accounts; and (3) bodies tell stories that people cannot or will not tell, either because they are unable, forbidden, or choose not to tell. Just as the proverbial "dead man's bones" do in fact tell tales, via forensic pathology and historical anthropometry, so too do our living bodies tell stories about our lives, whether or not these are ever consciously expressed. This glossary sketches some key concepts, definitions, and hypotheses relevant for using the construct of "embodiment" in epidemiological research, so as to promote not only rigorous science but also social equity in health. PMID- 15831682 TI - Health impact assessment: assessing opportunities and barriers to intersectoral health improvement in an expanded European Union. AB - On 1 May 2004 the European Union (EU) underwent unprecedented enlargement, from 15 to 25 countries, increasing its population by 20% to over 450 million. Although EU law has limited specific competence in the area of health, its influence on other policy sectors such as agriculture, trade, and employment has wide ranging implications for health. Yet with the exception of provisions on communicable disease control and food safety, public health considerations have played little part in negotiations on EU accession. This paper argues for an intersectoral public health approach in the expanded EU. It reviews the legal basis for assessing the health impacts of policy in the EU and, using health impact assessment as a case study, it examines how well the new member states may be prepared to tackle intersectoral public health action within the constraints imposed by EU policy. PMID- 15831683 TI - Quantitative health impact assessment: current practice and future directions. AB - STUDY OBJECTIVE: To assess what methods are used in quantitative health impact assessment (HIA), and to identify areas for future research and development. DESIGN: HIA reports were assessed for (1) methods used to quantify effects of policy on determinants of health (exposure impact assessment) and (2) methods used to quantify health outcomes resulting from changes in exposure to determinants (outcome assessment). MAIN RESULTS: Of 98 prospective HIA studies, 17 reported quantitative estimates of change in exposure to determinants, and 16 gave quantified health outcomes. Eleven (categories of) determinants were quantified up to the level of health outcomes. Methods for exposure impact assessment were: estimation on the basis of routine data and measurements, and various kinds of modelling of traffic related and environmental factors, supplemented with experts' estimates and author's assumptions. Some studies used estimates from other documents pertaining to the policy. For the calculation of health outcomes, variants of epidemiological and toxicological risk assessment were used, in some cases in mathematical models. CONCLUSIONS: Quantification is comparatively rare in HIA. Methods are available in the areas of environmental health and, to a lesser extent, traffic accidents, infectious diseases, and behavioural factors. The methods are diverse and their reliability and validity are uncertain. Research and development in the following areas could benefit quantitative HIA: methods to quantify the effect of socioeconomic and behavioural determinants; user friendly simulation models; the use of summary measures of public health, expert opinion and scenario building; and empirical research into validity and reliability. PMID- 15831684 TI - Impact of voluntary folate fortification on plasma homocysteine and serum folate in Australia from 1995 to 2001: a population based cohort study. AB - STUDY OBJECTIVE: To investigate the effect of the voluntary folate fortification policy in Australia on serum folate and total plasma homocysteine (tHcy) concentrations. DESIGN: Population based cohort study. SETTING: Perth, Western Australia. PARTICIPANTS: Men and women aged 27 to 77 years (n = 468), who were originally randomly selected from the Perth electoral roll. The cohort was surveyed in 1995/96 before widespread introduction of folate fortification of a variety of foods, and followed up on two occasions, firstly in 1998/99 and again in 2001, when a moderate number of folate fortified foods were available. Subjects with abnormal serum creatinine concentrations at baseline were excluded from this analysis. MAIN RESULTS: Repeated measures analysis of variance was used to determine changes in serum folate and tHcy over the three surveys and to assess whether time trends were related to age, sex, MTHFR C677T genotype, or consumption of folate fortified foods. An increase (38%) in mean serum folate (p < 0.0005) and a decrease (21%) in mean tHcy (p < 0.0005) were seen after introduction of the voluntary folate fortification policy in Australia. Serum folate was consistently higher (p = 0.032) and tHcy was consistently lower (p = 0.001) in subjects who consumed at least one folate fortified food compared with subjects who did not consume any folate fortified foods in the previous week. The time related changes in serum folate and tHcy were affected only by intake of folate fortified foods (p < 0.0005) and not by any other measured variables including age, sex, or MTHFR genotype. CONCLUSION: Voluntary fortification of foods with folate in Australia has been followed by a substantial increase in serum folate and decrease in tHcy in the general population. PMID- 15831685 TI - Work related and non-work related stress in relation to low leisure time physical activity in a Swedish population. PMID- 15831686 TI - Physical activity and health related quality of life among people with arthritis. AB - STUDY OBJECTIVE: To assess the association between physical activity and health related quality of life (HRQOL) among persons with arthritis or chronic joint symptoms (CJS). DESIGN: Cross sectional survey investigating the relation between physical activity level and HRQOL. HRQOL was estimated using the number of physically or mentally unhealthy days during the past 30 days. Physical activity was categorised as recommended, insufficient, or inactive according to federal activity recommendations. Persons with arthritis were defined as those with either self reported CJS or doctor diagnosed arthritis. SETTING: Community dwelling, US adults residing in all 50 states and the District of Columbia. PARTICIPANTS: Respondents (n = 212 000) in the 2001 behavioral risk factor surveillance system (BRFSS), an annual population based, telephone survey. MAIN RESULTS: The 33% of BRFSS respondents with arthritis had a mean of 6.7 physically and 4.9 mentally unhealthy days during the past 30 days, compared with 1.8 and 2.7 among those without arthritis. Inactive men and women were 1.2-2.4 times more likely to report impaired HRQOL compared with those who met physical activity recommendations. Men and women who engage in insufficient physical activity also report variably reduced HRQOL. CONCLUSIONS: Among people with arthritis, recommended levels of physical activity were associated with fewer mean physically and mentally unhealthy days and a decreased probability of having severely impaired physical or mental HRQOL. PMID- 15831687 TI - Late referral for assessment of renal failure. AB - It has been recommended that adult patients with a serum creatinine above 150 mumol/l should be referred to a nephrologist for specialist assessment. This study ascertained all patients in Northern Ireland with creatinine above this concentration in 2001 (n = 19 286 ) to see if this triggered referral within the subsequent year. After exclusion of those who were already known to a nephrologist and those who had acute renal failure, it was found that younger patients and diabetic patients were more likely to be referred. There was no difference in referral rates between male and female patients. However, only 6.5% of all non-diabetic subjects and 19% of diabetic patients were referred within 12 months after a first increased serum creatinine test. PMID- 15831688 TI - Relatively poor, absolutely ill? A study of regional income inequality in Russia and its possible health consequences. AB - STUDY OBJECTIVE: To investigate whether the income distribution in a Russian region has a "contextual" effect on individuals' self rated health, and whether the regional income distributions are related to regional health differences. METHODS: The Russia longitudinal monitoring survey (RLMS) is a survey (n = 7696) that is representative of the Russian population. With multilevel regressions both individual as well as contextual effects on self rated health were estimated. MAIN RESULTS: The effect of income inequality is not negative on men's self rated health as long as the level of inequality is not very great. When inequality levels are high, however, there is a tendency for men's health to be negatively affected. Regional health differences among men are in part explained by regional income differences. On the other hand, women do not seem to be affected in the same way, and individual characteristics like age and educational level seem to be more important. CONCLUSIONS: It seems that a rise in income inequality has no negative effect on men's self rated health as long as the level of inequality is not very great. On the other hand, when inequality levels are higher a rise tends to affect men's health negatively. A curvilinear relation between self rated health and income distribution is an interesting hypothesis. It could help to explain the confusing results that arise when you look at countries with a high degree of income inequality (USA) and those with lower income inequality (for example, Japan and New Zealand). PMID- 15831689 TI - Trends in smoking behaviour between 1985 and 2000 in nine European countries by education. AB - OBJECTIVE: To examine whether trends in smoking behaviour in Western Europe between 1985 and 2000 differed by education group. DESIGN: Data of smoking behaviour and education level were obtained from national cross sectional surveys conducted between 1985 and 2000 (a period characterised by intense tobacco control policies) and analysed for countries combined and each country separately. Annual trends in smoking prevalence and the quantity of cigarettes consumed by smokers were summarised for each education level. Education inequalities in smoking were examined at four time points. SETTING: Data were obtained from nine European countries: Norway, Sweden, Denmark, Finland, the United Kingdom, the Netherlands, Germany, Italy, and Spain. PARTICIPANTS: 451 386 non-institutionalised men and women 25-79 years old. MAIN OUTCOME MEASURES: Smoking status, daily quantity of cigarettes consumed by smokers. RESULTS: Combined country analyses showed greater declines in smoking and tobacco consumption among tertiary educated men and women compared with their less educated counterparts. In country specific analyses, elementary educated British men and women, and elementary educated Italian men showed greater declines in smoking than their more educated counterparts. Among Swedish, Finnish, Danish, German, Italian, and Spanish women, greater declines were seen among more educated groups. CONCLUSIONS: Widening education inequalities in smoking related diseases may be seen in several European countries in the future. More insight into effective strategies specifically targeting the smoking behaviour of low educated groups may be gained from examining the tobacco control policies of the UK and Italy over this period. PMID- 15831690 TI - Breast cancer survival in South Asian women in England and Wales. AB - STUDY OBJECTIVES: To estimate ethnic and socioeconomic differences in breast cancer incidence and survival between South Asians and non-South Asians in England and Wales, and to provide a baseline for surveillance of cancer survival in South Asians, the largest ethnic minority. SETTING: 115 712 women diagnosed with first primary invasive breast cancer in England and Wales during 1986-90 and followed up to 1995. METHODS/DESIGN: Ethnic group was ascribed by a computer algorithm on the basis of the name. Incidence rates were derived from 1991 census population denominators for each ethnic group. One and five year relative survival rates were estimated by age, quintile of material deprivation, and ethnic group, using national mortality rates to estimate expected survival. MAIN RESULTS: Age standardised incidence was 29% lower among South Asian women (40.5 per 100 000 per year) than among all other women (57.4 per 100 000). Five year age standardised relative survival was 70.3% (95%CI 65.2 to 75.4) for South Asian women and 66.7% (66.4 to 67.0) for other women. For both ethnic groups, survival was 8%-9% higher for women in the most affluent group than those in the most deprived group. In each deprivation category, however, survival was 3%-8% higher for South Asian women than other women. CONCLUSIONS: This national study confirms that breast cancer incidence is substantially lower in South Asians than other women in England and Wales. It also provides some evidence that South Asian women diagnosed up to 1990 had higher breast cancer survival than other women in England and Wales, both overall and in each category of deprivation. PMID- 15831691 TI - Midlife suicide risk, partner's psychiatric illness, spouse and child bereavement by suicide or other modes of death: a gender specific study. AB - STUDY OBJECTIVE: To describe gender specific suicide rates associated with partner's psychiatric disorder, loss of a spouse, or child by suicide or other causes, being a parent, and marital status. DESIGN: Nested case-control study. Information on causes of death, psychiatric admission, marital status, children, and socioeconomic factors was obtained from routine registers. SETTING: Denmark. PARTICIPANTS: 9011 people aged 25-60 years who committed suicide; 180 220 age gender matched controls; 111 172 marital partners; 174 672 children. MAIN RESULTS: The suicide risk in women whose partner had been first admitted with a psychiatric disorder after 31 December two years earlier was 6.9 (95% CI 3.6 to 13.0), whereas their male counterpart experienced a risk of 3.9 (2.7 to 5.6); p value gender difference = 0.39. Men who had lost their partner by suicide or other causes of death experienced a suicide risk of 46.2 (18.3 to 116.4) and 10.1 (6.5 to 15.8), respectively; the analogous risk among women were about one third: 15.8 (6.6 to 37.4) and 3.3 (1.5 to 7.2), respectively. Child bereavement by suicide or other causes imposed an approximate twofold risk increase in their parents, whereas being a parent was protective in women. Except for widows (1.6, 1.2 to 2.0) and widowers (3.0, 2.3 to 3.9) the suicide risk associated with being separated (2.0, 1.8 to 2.3), divorced (1.8, 1.7 to 2.0), never married (1.4, 1.3 to 1.6), cohabitant (1.2, 1.1 to 1.3) was virtually the same in the two sexes. CONCLUSIONS: The suicide risk is associated with partner psychiatric illness. Conjugal bereavement is particularly indicative of suicide in men, and spousal suicide is particularly indicative of suicide. Child bereavement is associated with parental suicide, while being a parent is protective against suicide in women. PMID- 15831692 TI - Physical functioning in elderly Europeans: 10 year changes in the north and south: the HALE project. AB - OBJECTIVES: To examine age related changes in physical functioning in elderly men and women. DESIGN: Prospective, population based study. SETTING: Population of 15 rural and urban centres in 10 European countries. PARTICIPANTS: Altogether 3496 men and women born between 1900 and 1920 who participated in the baseline survey of the HALE project in 1988-1991. The study population was examined again about five (in 1993-1995) and 10 (in 1999-2001) years after the baseline examination. MAIN OUTCOME MEASURES: Physical functioning was measured by means of a self administered questionnaire of activities of daily living (ADL). Dichotomised prevalence of disability and need for help in self care and mobility ADL were used as dependent variables in the analyses. RESULTS: Prevalence of disability and need for help tended to be higher in women than in men and in mobility abilities than in self care activities. Disability and need for help increased with advancing age but ameliorated over time from one birth cohort to another. In longitudinal analyses this beneficial time trend was independent of the effect of age, study, and region in self care disability in men and women (OR 0.85, 95% CI 0.75 to 0.97 and OR 0.64, 95% CI 0.43 to 0.97, respectively) and self care need for help in men (OR 0.83, 95% CI 0.70 to 0.96). Mobility disability among men and self care disability among women decreased more in the south than in the north. CONCLUSION: While European populations are aging, the proportions of elderly people with disability are decreasing. These results suggest that dynamics of functioning may differ across cultures. Future studies are needed to clarify which potentially modifiable and culturally determined factors protect against functional decline. PMID- 15831693 TI - Performance profile of an outcome measure: morbidity assessment index for newborns. AB - OBJECTIVE: Few tools have been optimised for use over the entire spectrum of neonatal morbidity and standardised for use in perinatal population and community health studies. The objective of this study was to determine the performance profile of the recently developed morbidity assessment index for newborns (MAIN score). This score was designed as a discriminative index of morbidity for the entire population of babies delivered >28 weeks gestation without a major congenital anomaly. DESIGN AND SETTING: MAIN score items were extracted retrospectively from the health records of 2892 consecutively born babies delivered beyond 28 weeks gestation in Edmonton area hospitals between June and December of 1999. MAIN RESULTS: The mean MAIN score in the general newborn population was 70.3 (95% confidence intervals 64.2 to 76.4). With the MAIN score tool, 84.6% of newborns scored from 0 to 150 (no/minimal morbidity), 11.3% from 151 to 500 (mild), 3.1% from 501 to 800 (moderate), and 1% had >800 (severe) score. The MAIN score tool was sufficiently sensitive to detect significant effects of low gestational age, low birth weight, male sex, caesarean delivery, tertiary hospital delivery, twins/triplets, non-vertex presentation, prenatal illicit drug use, and medical complications of pregnancy. CONCLUSION: The MAIN score fulfills the need for a simple, universal, yet sensitive and robust tool to provide a numerical index of early neonatal outcomes of prenatal care and adverse prenatal exposures in babies delivered beyond 28 weeks gestation. The performance of the MAIN score agrees well with the current medical awareness regarding the impact of adverse prenatal exposures on newborn morbidity. PMID- 15831694 TI - Are international differences in the outcomes of acute coronary syndromes apparent or real? A multilevel analysis. AB - STUDY OBJECTIVE: International variation in the outcomes of patients with acute coronary syndromes (ACS) has been well reported. The relative contributions of patient, hospital, and country level factors on clinical outcomes, however, remain unclear, and thus, was the objective of this study. DESIGN: Multilevel logistic regression models were developed for death/(re)infarction (MI) at 30 days and death in one year, with patients (1st level) nested in hospitals (2nd level) and hospitals in countries (3rd level). SETTINGS: The GUSTO IV ACS clinical trial was carried out at 458 hospital sites in 24 countries. PATIENTS: 7800 non-ST segment elevation (NSTE) ACS patients. MAIN RESULTS: There were substantial variations among countries in the processes and outcomes of care at 30 days, ranging from 5.4% to 50.0% for percutaneous coronary intervention, 4.3% to 21.2% for coronary artery bypass graft surgery, 5.0% to 13.9% for 30 day death/(re)MI, and 4.9% to 14.8% for one year mortality. However, the residual inter-country variations in 30 day death/(re)MI and one year mortality became non significant and nearly disappeared (p > 0.500 for both) after adjusting for key baseline patient characteristics and hospital factors, which became significant (p < 0.01 for both). Patient level factors accounted for 96%-99% of total variation in these end points, leaving the remaining 1% and 4% of variance attributable to hospital level factors. CONCLUSION: The international differences in clinical outcomes in this study of NSTE ACS are primarily accounted for by the patient level factors, with hospital level factors playing a minor part, and the country level factors a negligible one. These findings have significant policy and research implications involving international collaboration and comparisons. PMID- 15831695 TI - Bidirectional associations between depression/anxiety and bowel disease in a population based cohort. PMID- 15831696 TI - Failure to identify association between deprivation and incidence of lung cancer surprising. PMID- 15831697 TI - Acute myeloid leukemia bearing cytoplasmic nucleophosmin (NPMc+ AML) shows a distinct gene expression profile characterized by up-regulation of genes involved in stem-cell maintenance. AB - Approximately one third of acute myeloid leukemias (AMLs) are characterized by aberrant cytoplasmic localization of nucleophosmin (NPMc+ AML), consequent to mutations in the NPM putative nucleolar localization signal. These events are mutually exclusive with the major AML-associated chromosomal rearrangements, and are frequently associated with normal karyotype, FLT3 mutations, and multilineage involvement. We report the gene expression profiles of 78 de novo AMLs (72 with normal karyotype; 6 without major chromosomal abnormalities) that were characterized for the subcellular localization and mutation status of NPM. Unsupervised clustering clearly separated NPMc+ from NPMc- AMLs, regardless of the presence of FLT3 mutations or non-major chromosomal rearrangements, supporting the concept that NPMc+ AML represents a distinct entity. The molecular signature of NPMc+ AML includes up-regulation of several genes putatively involved in the maintenance of a stem-cell phenotype, suggesting that NPMc+ AML may derive from a multipotent hematopoietic progenitor. PMID- 15831698 TI - Nonhemolytic antibody-induced loss of erythrocyte surface antigen. AB - Transfusion of red blood cells (RBCs) into patients with anti-donor RBC antibodies (crossmatch-incompatible transfusion) can result in lethal antibody mediated hemolysis. Less well appreciated is the ability of anti-RBC antibodies to specifically remove their target antigen from donor RBCs without compromising cell survival or adversely affecting the transfusion recipient. In an effort to elucidate the mechanistic details of this process, we describe the first animal model of nonhemolytic antibody-induced RBC antigen loss. RBCs from transgenic mHEL mice express surface hen egg lysozyme (HEL) as a transmembrane protein. Transfusion of mHEL RBCs into mice immunized with HEL results in selective loss of HEL antigen from donor RBCs without affecting other blood group antigens or reducing the circulatory life span of the transfused RBCs. While this process does not require the presence of a spleen, it requires both anti-RBC immunoglobulin G (IgG) antibodies and the FcgammaIII receptor. These studies provide mechanistic insight into the phenomenon of antigen loss during incompatible transfusion in humans. PMID- 15831699 TI - Crystal structure of the Jak3 kinase domain in complex with a staurosporine analog. AB - Jak (Janus kinase) family nonreceptor tyrosine kinases are central mediators of cytokine signaling. The Jak kinases exhibit distinct cytokine receptor association profiles and so transduce different signals. Jak3 expression is limited to the immune system, where it plays a key role in signal transduction from cytokine receptors containing the common gamma-chain, gammac. Patients unable to signal via gammac present with severe combined immunodeficiency (SCID). The finding that Jak3 mutations result in SCID has made it a target for development of lymphocyte-specific immunosuppressants. Here, we present the crystal structure of the Jak3 kinase domain in complex with staurosporine analog AFN941. The kinase domain is in the active conformation, with both activation loop tyrosine residues phosphorylated. The phosphate group on pTyr981 in the activation loop is in part coordinated by an arginine residue in the regulatory C helix, suggesting a direct mechanism by which the active position of the C-helix is induced by phosphorylation of the activation loop. Such a direct coupling has not been previously observed in tyrosine kinases and may be unique to Jak kinases. The crystal structure provides a detailed view of the Jak3 active site and will facilitate computational and structure-directed approaches to development of Jak3-specific inhibitors. PMID- 15831700 TI - Resistance to hepcidin is conferred by hemochromatosis-associated mutations of ferroportin. AB - Ferroportin (FPN) mediates iron export from cells; FPN mutations are associated with the iron overloading disorder hemochromatosis. Previously, we found that the A77D, V162del, and G490D mutations inhibited FPN activity, but that other disease associated FPN variants retained full iron export capability. The peptide hormone hepcidin inhibits FPN as part of a homeostatic negative feedback loop. We measured surface expression and function of wild-type FPN and fully active FPN mutants in the presence of hepcidin. We found that the Y64N and C326Y mutants of FPN are completely resistant to hepcidin inhibition and that N144D and N144H are partially resistant. Hemochromatosis-associated FPN mutations, therefore, either reduce iron export ability or produce an FPN variant that is insensitive to hepcidin. The former mutation type is associated with Kupffer-cell iron deposition and normal transferrin saturation in vivo, whereas patients with the latter category of FPN mutation have high transferrin saturation and tend to deposit iron throughout the liver parenchyma. FPN-linked hemochromatosis may have a variable pathogenesis depending on the causative FPN mutant. PMID- 15831701 TI - IL-6 blocks a discrete early step in lymphopoiesis. AB - Animals lacking Src homology 2 domain-containing inositol 5-phosphatase (SHIP) display a reduction in lymphopoiesis and a corresponding enhancement of myelopoiesis. These effects are mediated at least in part by elevated levels of interleukin 6 (IL-6). Here, we show the lymphopoiesis block in SHIP-/- mice is due to suppression of the lymphoid lineage choice by uncommitted progenitors. The suppression can be reproduced in vitro with recombinant IL-6, and IL-6 acts directly on hematopoietic progenitors. The block is partially overcome in SHIP-/- IL-6-/- double-deficient animals. IL-6 does not suppress but actually enhances proliferation of lymphoid-committed progenitors, indicating the IL-6 target cells are hematopoietic stem cells or multipotent progenitors. The findings suggest a mechanism for the lymphopenia that accompanies proinflammatory diseases. PMID- 15831702 TI - CD144 (VE-cadherin) is transiently expressed by fetal liver hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) and endothelial progenitors arise from a common embryonic precursor. However, these populations diverge prior to the onset of definitive hematopoiesis, as HSCs become CD45+ and are thought to lose the expression of endothelial markers. After the onset of definitive hematopoiesis, CD144 (vascular endothelial [VE]-cadherin) has been considered a specific marker of endothelial cells. In contrast, we found that virtually all HSC activity from embryonic day 13.5 (E13.5) fetal liver was CD144+. CD144 expression declined on E16.5 fetal liver HSCs and was absent from adult bone marrow HSCs. This identified a new marker that is differentially expressed between fetal and adult HSCs, and enhanced the purification of HSCs from the E13.5 fetal liver. These results emphasize the close developmental relationship between hematopoietic and endothelial cells, while indicating that CD144 is not a specific marker of endothelial cells during fetal development. PMID- 15831703 TI - Microcytic anemia, erythropoietic protoporphyria, and neurodegeneration in mice with targeted deletion of iron-regulatory protein 2. AB - Iron-regulatory proteins (IRPs) 1 and 2 posttranscriptionally regulate expression of transferrin receptor (TfR), ferritin, and other iron metabolism proteins. Mice with targeted deletion of IRP2 overexpress ferritin and express abnormally low TfR levels in multiple tissues. Despite this misregulation, there are no apparent pathologic consequences in tissues such as the liver and kidney. However, in the central nervous system, evidence of abnormal iron metabolism in IRP2-/- mice precedes the development of adult-onset progressive neurodegeneration, characterized by widespread axonal degeneration and neuronal loss. Here, we report that ablation of IRP2 results in iron-limited erythropoiesis. TfR expression in erythroid precursors of IRP2-/- mice is reduced, and bone marrow iron stores are absent, even though transferrin saturation levels are normal. Marked overexpression of 5-aminolevulinic acid synthase 2 (Alas2) results from loss of IRP-dependent translational repression, and markedly increased levels of free protoporphyrin IX and zinc protoporphyrin are generated in IRP2-/- erythroid cells. IRP2-/- mice represent a new paradigm of genetic microcytic anemia. We postulate that IRP2 mutations or deletions may be a cause of refractory microcytic anemia and bone marrow iron depletion in patients with normal transferrin saturations, elevated serum ferritins, elevated red cell protoporphyrin IX levels, and adult-onset neurodegeneration. PMID- 15831704 TI - Production and characterization of erythropoietic protoporphyric heterodimeric ferrochelatases. AB - Mutations resulting in diminished activity of the dimeric enzyme ferrochelatase are a prerequisite for the inherited disorder erythropoietic protoporphyria (EPP). Patients with clinical EPP have only 10% to 30% of normal levels of ferrochelatase activity, and although many patients with EPP have one mutant allele and one "low-expression" normal allele, the possibility remains that, for some, low ferrochelatase activity may result from an EPP mutation that has an impact on both subunits of the wild-type/mutant heterodimer. Here we present data for 12 ferrochelatase wild-type/EPP mutant heterodimers showing that some mutations result in heterodimers with the residual activity anticipated from individual constituents, whereas others result in heterodimers with significantly lower activity than would be predicted. Although the data do not allow an a priori prediction of heterodimeric residual activity based solely on the in vitro activity of EPP homodimers or the position of the mutated residue within ferrochelatase, mutations that affect the dimer interface or [2Fe-2S] cluster have a significantly greater impact on residual activity than would be predicted. These data suggest that some EPP mutations may result in clinically overt EPP in the absence of a low-expression, wild-type allele; this is of potential significance for genetic counseling of patients with EPP. PMID- 15831705 TI - Hematopoietic differentiation of human embryonic stem cells progresses through sequential hematoendothelial, primitive, and definitive stages resembling human yolk sac development. AB - We elucidate the cellular and molecular kinetics of the stepwise differentiation of human embryonic stem cells (hESCs) to primitive and definitive erythromyelopoiesis from human embryoid bodies (hEBs) in serum-free clonogenic assays. Hematopoiesis initiates from CD45 hEB cells with emergence of semiadherent mesodermal-hematoendothelial (MHE) colonies that can generate endothelium and form organized, yolk sac-like structures that secondarily generate multipotent primitive hematopoietic stem progenitor cells (HSPCs), erythroblasts, and CD13+CD45+ macrophages. A first wave of hematopoiesis follows MHE colony emergence and is predominated by primitive erythropoiesis characterized by a brilliant red hemoglobinization, CD71/CD325a (glycophorin A) expression, and exclusively embryonic/fetal hemoglobin expression. A second wave of definitive-type erythroid burst-forming units (BFU-e's), erythroid colony forming units (CFU-e's), granulocyte-macrophage colony-forming cells (GM-CFCs), and multilineage CFCs follows next from hEB progenitors. These stages of hematopoiesis proceed spontaneously from hEB-derived cells without requirement for supplemental growth factors during hEB differentiation. Gene expression analysis of differentiating hEBs revealed that initiation of hematopoiesis correlated with increased levels of SCL/TAL1, GATA1, GATA2, CD34, CD31, and the homeobox gene-regulating factor CDX4 These data indicate that hematopoietic differentiation of hESCs models the earliest events of embryonic and definitive hematopoiesis in a manner resembling human yolk sac development, thus providing a valuable tool for dissecting the earliest events in human HSPC genesis. PMID- 15831706 TI - Negative regulation of platelet function by a secreted cell repulsive protein, semaphorin 3A. AB - Semaphorin 3A (Sema3A) is a secreted disulfide-bound homodimeric molecule that induces growth cone collapse and repulsion of axon growth in the nervous system. Recently, it has been demonstrated that Sema3A is produced by endothelial cells and inhibits integrin function in an autocrine fashion. In this study, we investigated the effects of Sema3A on platelet function by using 2 distinct human Sema3A chimera proteins. We detected expression of functional Sema3A receptors in platelets and dose-dependent and saturable binding of Sema3A to platelets. Sema3A dose-dependently inhibited activation of integrin alphaIIbbeta3 by all agonists examined including adenosine diphosphate (ADP), thrombin, convulxin, phorbol 12 myristate 13-acetate, and A23187. Sema3A inhibited not only platelet aggregation induced by thrombin or collagen but also platelet adhesion and spreading on immobilized fibrinogen. Moreover, Sema3A impaired alphaIIbbeta3-independent spreading on glass coverslips and aggregation-independent granular secretion. Sema3A inhibited agonist-induced elevation of filamentous action (F-actin) contents, phosphorylation of cofilin, and Rac1 activation. In contrast, Sema3A did not affect the levels of cyclic nucleotides or agonist-induced increase of intracellular Ca2+ concentrations. Thus, the extensive inhibition of platelet function by Sema3A appears to be mediated, at least in part, through impairment of agonist-induced Rac1-dependent actin rearrangement. PMID- 15831707 TI - Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV. AB - Viral heterogeneity is a major hurdle for potential therapeutic use of RNA interference (RNAi) against HIV-1. To determine the extent of RNAi tolerance to mutations, we tested 3 viral target sites with differing propensity for mutations: a highly variable rev sequence, a gag sequence conserved only among clade B isolates, and a vif sequence highly conserved across clades. Lentiviral expression of all 3 shRNAs inhibited replication of the homologous HIV(IIIB) strain. However, they differed in their ability to protect primary CD4 T cells against multiple isolates within and across HIV clades. The least conserved rev sequence inhibited only 2 of 5 clade B isolates. The gag sequence (conserved within clade B) protected 5 of 5 clade B isolates but not other clade viruses with 2 or 3 mutations in the central region. In contrast, the vif sequence, which was conserved across clades except for single mutations at positions 14 and 17, inhibited viruses from 5 different clades. Moreover, siRNAs with introduced mutations at sites of gag sequence polymorphisms showed reduced antiviral activity, whereas mutations in vif siRNA only modestly decreased silencing. Thus, although 1 or 2 mutations at peripheral sites are tolerated, mutations in the central target cleavage region abolish RNAi activity. PMID- 15831708 TI - Repressible transgenic model of NRAS oncogene-driven mast cell disease in the mouse. AB - To create a model in which to study the effects of RAS dysregulation in hematopoietic disease, we developed separate founder lines of transgenic mice, with the tetracycline transactivator (tTA) driven by the Vav hematopoietic promoter in one line and NRASV12 driven by the tetracycline responsive element (TRE2) in the other. When these lines are crossed, doubly transgenic animals uniformly develop a disease similar to human aggressive systemic mastocytosis (ASM) or mast cell leukemia (MCL) when they are between 2 and 4 months of age. Disease is characterized by tissue infiltrates of large, well-differentiated mast cells in the spleen, liver, skin, lung, and thymus. Analysis of bone sections shows small to large foci of similarly well-differentiated mast cells. Results also show that transgene expression and diseases are repressible through the administration of doxycycline in the drinking water of affected animals, indicating that NRASV12 expression is required to initiate and maintain disease in doubly transgenic mice. Our inducible system of transgenes, developed as a model of mutant NRASV12 oncogene-driven myeloid disease, will be useful for studying the role of RAS dysregulation in hematopoietic disease in general and in discrete human diseases, specifically ASM and MCL. PMID- 15831709 TI - Engagement of specific T-cell surface molecules regulates cytoskeletal polarization in HTLV-1-infected lymphocytes. AB - Cell-cell contact is required for efficient transmission of human T-lymphotropic virus type 1 (HTLV-1). An HTLV-1-infected cell polarizes its microtubule organizing center (MTOC) toward the cell-cell junction; HTLV-1 core (Gag) complexes and the HTLV-1 genome accumulate at the point of contact and are then transferred to the uninfected cell. However, the mechanisms involved in this cytoskeletal polarization and transport of HTLV-1 complexes are unknown. Here, we tested the hypothesis that engagement of a specific T-cell surface ligand is synergistic with HTLV-1 infection in causing polarization of the MTOC to the cell contact region. We show that antibodies to intercellular adhesion molecule-1 (ICAM-1; CD54) caused MTOC polarization at a higher frequency in HTLV-1-infected cells. ICAM-1 is upregulated on HTLV-1-infected cells, and, in turn, ICAM-1 on the cell surface upregulates HTLV-1 gene expression. We propose that a positive feedback loop involving ICAM-1 and HTLV-1 Tax protein facilitates the formation of the virologic synapse and contributes to the T-cell tropism of HTLV-1. In contrast, MTOC polarization induced in T cells by antibodies to CD3 or CD28 was significantly inhibited by HTLV-1 infection. PMID- 15831710 TI - Differentiation-dependent activation of the human intestinal alkaline phosphatase promoter by HNF-4 in intestinal cells. AB - The intestinal alkaline phosphatase gene (ALPI) encodes a digestive brush-border enzyme, which is highly upregulated during small intestinal epithelial cell differentiation. To identify new putative promoter motifs responsible for the regulation of ALPI expression during differentiation of the enterocytes, we have conducted a computer-assisted cis-element search of the proximal human ALPI promoter sequence. A putative recognition site for the transcription factor hepatocyte nuclear factor (HNF)-4 was predicted at the positions from -94 to -82 in relation to the translational start site. The ability of HNF-4alpha to stimulate the expression from the ALPI promoter was investigated in the nonintestinal Hela cell line. Cotransfection with an HNF-4alpha expression vector demonstrated a direct activation of the ALPI promoter through this -94 to -82 element. EMSA showed that HNF-4alpha from nuclear extracts of differentiated intestinal epithelial cells (Caco-2) bound with high affinity to the predicted HNF-4 binding site. A 521 bp promoter fragment containing the HNF-4 binding site demonstrated a differentiation-dependent increase in promoter activity in Caco-2 cells. The presence of the HNF-4 binding site was necessary for this increase to occur. PMID- 15831711 TI - Modulation of electroneutral Na transport in sheep rumen epithelium by luminal ammonia. AB - Ammonia is an abundant fermentation product in the forestomachs of ruminants and the intestine of other species. Uptake as NH3 or NH4+ should modulate cytosolic pH and sodium-proton exchange via Na+/H+ exchanger (NHE). Transport rates of Na+, NH4+, and NH3 across the isolated rumen epithelium were studied at various luminal ammonia concentrations and pH values using the Ussing chamber method. The patch-clamp technique was used to identify an uptake route for NH4+. The data show that luminal ammonia inhibits electroneutral Na transport at pH 7.4 and abolishes it at 30 mM (P < 0.05). In contrast, at pH 6.4, ammonia stimulates Na transport (P < 0.05). Flux data reveal that at pH 6.4, approximately 70% of ammonia is absorbed in the form of NH4+, whereas at pH 7.4, uptake of NH3 exceeds that of NH4+ by a factor of approximately four. The patch-clamp data show a quinidine-sensitive permeability for NH4+ and K+ but not Na+. Conductance was 135 +/- 12 pS in symmetrical NH(4)Cl solution (130 mM). Permeability was modulated by the concentration of permeant ions, with P(K) > P(NH4) at high and P(NH4) > P(K) at lower external concentrations. Joint application of both ions led to anomalous mole fraction effects. In conclusion, the luminal pH determines the predominant form of ammonia absorption from the rumen and the effect of ammonia on electroneutral Na transport. Protons that enter the cytosol through potassium channels in the form of NH4+ stimulate and nonionic diffusion of NH3 blocks NHE, thus contributing to sodium transport and regulation of pH. PMID- 15831712 TI - Interleukin-1 inhibits the induction of insulin-like growth factor-I by growth hormone in CWSV-1 hepatocytes. AB - Sepsis results in hepatic "growth hormone (GH) resistance" with reductions in plasma IGF-I despite a two- to fourfold increase in circulating GH. In this study, we examine the effects of IL-1 on GH receptor (GHR) expression, GH signaling (via the JAK/STAT and MAPK pathways), and the induction of gene expression [IGF-I mRNA and serine protease inhibitor (Spi) 2.1] by GH in CWSV-1 hepatocytes. Incubation of cells with IL-1beta (10 ng/ml, 24 h) had no effect on the relative abundance of GHR or signaling proteins JAK2, STAT5b, and ERK1/2 in cell lysates. Baseline phosphorylation of GHR, JAK2, STAT5b, and ERK1/2 was minimal. After GH stimulation, tyrosine phosphorylation of GHR, JAK2, STAT5b, and ERK1/2 increased 2- to 10-fold. However, neither the time course nor the magnitude of GHR, JAK2, and ERK1/2 phosphorylation by GH were significantly altered by IL-1. The GH-induced translocation of STAT5b to the nucleus was not prevented by IL-1. Although phosphorylated STAT5 in nuclear extracts from GH + IL 1 cells was decreased by 24% (vs. controls) 15 min after GH stimulation, this did not result in reduced STAT5-DNA binding activity. Pretreatment with IL-1 did not significantly decrease IGF-I mRNA stability. We conclude that IL-1 only minimally affects the time course of JAK2/STAT5 and MAPK signaling by GH. Therefore, an inhibitory effect of IL-1 on IGF-I and Spi 2.1 mRNA synthesis by GH represents the most likely mechanism for IL-1-mediated GH resistance. PMID- 15831713 TI - Suppressor of cytokine signaling-2 modulates the fibrogenic actions of GH and IGF I in intestinal mesenchymal cells. AB - Growth hormone (GH) and IGF-I play important roles in wound healing during intestinal injury and inflammation, but there is also indirect evidence that locally expressed IGF-I may act to induce excessive collagen deposition, which can lead to intestinal fibrosis. Factors that dictate the balance between normal wound healing and excessive healing responses are unknown. Using RNase protection assay and in situ hybridization, we determined whether GH and/or IGF-I increase type I collagen deposition in the intestine of rats fed by total parenteral nutrition (TPN), a feeding modality used for many patients following intestinal surgery and resection. We also used an in vitro model system to confirm our in vivo effects and to directly evaluate the relative potency of GH and IGF-I on DNA synthesis and collagen deposition in intestinal myofibroblasts. Both GH and IGF-I stimulated collagen production in vivo and in vitro, and IGF-I, but not GH, stimulated DNA synthesis in vitro. In collagen production, GH was less potent than IGF-I. Suppressors of cytokine signaling (SOC) are cytokine-inducible proteins that negatively feedback to inhibit the actions of cytokines and we recently found that GH selectively upregulates SOC-2 in the intestine of TPN-fed rats. We examined whether SOC-2 may be responsible for the difference in magnitude of action of GH and IGF-I on collagen accumulation. GH, but not IGF-I, induced SOC-2 in isolated myofibroblasts, and overexpression of SOC-2 led to a suppression of GH- and IGF-I-induced collagen accumulation. SOC-2 null mice infused with IGF-I showed greater collagen gene expression compared with wild type (WT) mice. Myofibroblasts isolated from SOC-2 null mice showed increased IGF I-stimulated DNA synthesis compared with WT cells. Taken together, these findings suggest that SOC-2 induced by GH may play an important role in suppressing collagen accumulation and mesenchymal cell proliferation induced by GH or GH induced IGF-I, providing a mechanism for the differing potencies of GH and IGF-I on intestinal mesenchyme and collagen synthesis. PMID- 15831714 TI - Inhibition of transient lower esophageal sphincter relaxations by electrical acupoint stimulation. AB - Acupuncture has been shown to modulate visceral sensation and function. Traditionally, stimulation at the Neiguan (pericardial meridian) has been used to treat upper gastrointestinal symptoms. Some of the effects of acupuncture may be mediated through release of endogenous opioids and are reversed by naloxone. Gastric distension is the major trigger for transient lower esophageal sphincter (LES) relaxations (TLESRs). The aim of this study was to investigate the effect of electric stimulation at the Neiguan and naloxone on the TLESRs. In 14 healthy volunteers, electrical acupoint stimulation was applied at the Neiguan and a sham point on the hip in randomized order on the same day. In 12 healthy volunteers, the effects of naloxone (80 microg/kg iv bolus injection) and saline on electrical acupoint stimulation were compared on separate days at least 1 wk apart. Esophageal motility was measured during distension of the proximal stomach with 500 ml of air using a barostat balloon. Electric acupoint stimulation at the Neiguan decreased the rate of TLESRs by approximately 40% from a median of 6/h to 3.5/h (P < 0.02). Acupoint stimulation had no effect on basal LES pressure, the residual LES pressure during TLESRs, the duration of TLESRs, or gastrointestinal symptoms of fullness, bloating, discomfort, or nausea. The effect of acupoint stimulation was not inhibited by naloxone. Electric acupoint stimulation at the Neiguan significantly inhibits the frequency of TLESRs in response to gastric distention in healthy subjects. This effect does not appear to be mediated through mu-opioid receptors. PMID- 15831715 TI - Differential proteome analysis of aging in rat skeletal muscle. AB - To identify the mechanisms underlying muscle aging, we have undertaken a high resolution differential proteomic analysis of gastrocnemius muscle in young adults, mature adults, and old LOU/c/jall rats. Two-dimensional gel electrophoresis and subsequent MALDI-ToF mass spectrometry analyses led to the identification of 40 differentially expressed proteins. Strikingly, most differences characterized old (30-month) animals, whereas young (7-month) and mature (18-month) adults exhibited similar patterns of expression. Important modifications in contractile (actin, myosin light-chains, troponins-T) and cytoskeletal (desmin, tubulin) proteins, and in essential regulatory proteins (gelsolin, myosin binding proteins, CapZ-beta, P23), likely account for dysfunctions in old muscle force generation and speed of contraction. Other features support decreases in cytosolic (triose-phosphate isomerase, enolase, glycerol-3-P dehydrogenase, creatine kinase) and mitochondrial (isocitrate dehydrogenase, cytochrome-c oxidase) energy metabolisms. Muscle aging is often associated with increased oxidative stress. Accordingly, we observed differential regulation of molecular chaperones (hsp20, hsp27, reticuloplasmin ER60) and of proteins implicated in reactive aldehyde detoxification (aldehyde dehydrogenase, glutathione transferase, glyoxalase). We further noticed up-regulation of proteins involved in transcriptional elongation (RNA capping protein) and RNA editing (Apobec2). Most of these proteins were previously unrecognized as differentially expressed in old muscles, and they represent novel starting points for elucidating the mechanisms of muscle aging. PMID- 15831716 TI - Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection. AB - Ebola virus (EboV) causes rapidly fatal hemorrhagic fever in humans and there is currently no effective treatment. We found that the infection of African green monkey kidney (Vero) cells by vesicular stomatitis viruses bearing the EboV glycoprotein (GP) requires the activity of endosomal cysteine proteases. Using selective protease inhibitors and protease-deficient cell lines, we identified an essential role for cathepsin B (CatB) and an accessory role for cathepsin L (CatL) in EboV GP-dependent entry. Biochemical studies demonstrate that CatB and CatL mediate entry by carrying out proteolysis of the EboV GP subunit GP1 and support a multistep mechanism that explains the relative contributions of these enzymes to infection. CatB and CatB/CatL inhibitors diminish the multiplication of infectious EboV-Zaire in cultured cells and may merit investigation as anti EboV drugs. PMID- 15831717 TI - Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. AB - Microglial cells represent the immune system of the mammalian brain and therefore are critically involved in various injuries and diseases. Little is known about their role in the healthy brain and their immediate reaction to brain damage. By using in vivo two-photon imaging in neocortex, we found that microglial cells are highly active in their presumed resting state, continually surveying their microenvironment with extremely motile processes and protrusions. Furthermore, blood-brain barrier disruption provoked immediate and focal activation of microglia, switching their behavior from patroling to shielding of the injured site. Microglia thus are busy and vigilant housekeepers in the adult brain. PMID- 15831719 TI - An observation of PKJKP: inferences on inner core shear properties. AB - The seismic phase PKJKP, which traverses the inner core as a shear wave and would provide direct evidence for its solidity, has been difficult to detect. Using stacked broadband records from the Grafenberg array in Germany, we documented a high signal-to-noise phase, the arrival time and slowness of which agree with theoretical predictions for PKJKP. The back azimuth of this arrival is also consistent with predictions for PKJKP, as is the comparison with a pseudoliquid inner core model. Envelope modeling of the PKJKP waveform implies a shear velocity gradient with depth in the inner core that is slightly larger than that in the preliminary reference Earth model. PMID- 15831720 TI - In search of a lifeline. PMID- 15831721 TI - Stem cells. Restiveness grows at NIH over Bush research restrictions. PMID- 15831722 TI - AIDS research. IOM panel clears HIV prevention study. PMID- 15831723 TI - Nuclear waste. Academy gets the word out after tussle with agency. PMID- 15831724 TI - Geochemistry. Gasping for air in Permian hard times. PMID- 15831726 TI - Japan. Space vision backs peer-reviewed science. PMID- 15831725 TI - European science. A second entry in the Mars sweepstakes. PMID- 15831727 TI - Particle physics. Magnetic scope angles for axions. PMID- 15831718 TI - Diversity of the human intestinal microbial flora. AB - The human endogenous intestinal microflora is an essential "organ" in providing nourishment, regulating epithelial development, and instructing innate immunity; yet, surprisingly, basic features remain poorly described. We examined 13,355 prokaryotic ribosomal RNA gene sequences from multiple colonic mucosal sites and feces of healthy subjects to improve our understanding of gut microbial diversity. A majority of the bacterial sequences corresponded to uncultivated species and novel microorganisms. We discovered significant intersubject variability and differences between stool and mucosa community composition. Characterization of this immensely diverse ecosystem is the first step in elucidating its role in health and disease. PMID- 15831728 TI - Population genetics. Private partnership to trace human history. PMID- 15831730 TI - Asian tsunamis. Model shows islands muted tsunami after latest Indonesian quake. PMID- 15831729 TI - Toxicology. EPA kills Florida pesticide study. PMID- 15831732 TI - European research. A framework for change? PMID- 15831731 TI - Infectious diseases. Veterinary scientists shore up defenses against bird flu. PMID- 15831733 TI - European research. E.U. wins over researchers with a ticket to ride. PMID- 15831734 TI - European research. The dos and don'ts of getting an E.U. grant. PMID- 15831735 TI - Addiction research. Ibogaine therapy: a 'vast, uncontrolled experiment'. PMID- 15831737 TI - Superconductivity. New wave of electrical wires inches closer to market. PMID- 15831736 TI - Ecology. Experimental drought predicts grim future for rainforest. PMID- 15831738 TI - The problem with patents. PMID- 15831739 TI - Defining the concept of public information. PMID- 15831740 TI - PLOS position on NIH public access policy. PMID- 15831741 TI - Comment on "Grain boundary-mediated plasticity in nanocrystalline nickel". PMID- 15831743 TI - Ecology. Importing timber, exporting ecological impact. PMID- 15831744 TI - Ocean science. Coral clues to rapid sea-level change. AB - Earth's climate can change substantially on time scales of 1000 years or so, but given the time it takes for an ice sheet to grow or melt, it has been unclear whether continental ice sheets-and hence global sea levels-mirror these rapid changes. In his Perspective, Henderson discusses the report by Thompson and Goldstein, who have used a new correction method to date coral samples that are up to 250,000 years old. The corals can be used to deduce past sea levels. The resulting sea-level record shows that sea levels have varied on millennial time scales even during times of high sea level and relative climate stability. PMID- 15831746 TI - Evolution. Life on the early Earth: a sedimentary view. PMID- 15831745 TI - Cell biology. Of grainy heads and broken skins. PMID- 15831747 TI - Chemistry. A new route to designer antibiotics. PMID- 15831748 TI - Unveiling the mechanisms of cell-cell fusion. AB - Cell-cell fusion is fundamental to the development and physiology of multicellular organisms, but little is known of its mechanistic underpinnings. Recent studies have revealed that many proteins involved in cell-cell fusion are also required for seemingly unrelated cellular processes such as phagocytosis, cell migration, axon growth, and synaptogenesis. We review advances in understanding cell-cell fusion by contrasting it with virus-cell and intracellular vesicle fusion. We also consider how proteins involved in general aspects of membrane dynamics have been co-opted to control fusion of diverse cell types by coupling with specialized proteins involved in cell-cell recognition, adhesion, and signaling. PMID- 15831749 TI - A pair of shelled eggs inside a female dinosaur. AB - An oviraptosaurian specimen (Dinosauria, Theropoda) from an Upper Cretaceous formation in China retains a pair of shelled eggs in the pelvis, providing direct evidence that oviraptorosaurian dinosaurs laid paired elongatoolithid eggs. The presence of the paired eggs suggests that theropod dinosaurs had two functional oviducts (like crocodiles) but that each oviduct produced only one egg at a time and that an entire egg clutch was laid through multiple ovipositions (like birds). The orientations of the eggs inside the skeleton and in clutches indicate that the mother came to the center of the nest to lay eggs. PMID- 15831750 TI - Impact of humans on the flux of terrestrial sediment to the global coastal ocean. AB - Here we provide global estimates of the seasonal flux of sediment, on a river-by river basis, under modern and prehuman conditions. Humans have simultaneously increased the sediment transport by global rivers through soil erosion (by 2.3 +/ 0.6 billion metric tons per year), yet reduced the flux of sediment reaching the world's coasts (by 1.4 +/- 0.3 billion metric tons per year) because of retention within reservoirs. Over 100 billion metric tons of sediment and 1 to 3 billion metric tons of carbon are now sequestered in reservoirs constructed largely within the past 50 years. African and Asian rivers carry a greatly reduced sediment load; Indonesian rivers deliver much more sediment to coastal areas. PMID- 15831751 TI - An epidermal barrier wound repair pathway in Drosophila is mediated by grainy head. AB - We used wounded Drosophila embryos to define an evolutionarily conserved pathway for repairing the epidermal surface barrier. This pathway includes a wound response enhancer from the Ddc gene that requires grainy head (grh) function and binding sites for the Grh transcription factor. At the signaling level, tyrosine kinase and extracellular signal-regulated kinase (ERK) activities are induced in epidermal cells near wounds, and activated ERK is required for a robust wound response. The conservation of this Grh-dependent pathway suggests that the repair of insect cuticle and mammal skin is controlled by an ancient, shared control system for constructing and healing the animal body surface barrier. PMID- 15831752 TI - Quantum phase transition of a magnet in a spin bath. AB - The excitation spectrum of a model magnetic system, LiHoF4, was studied with the use of neutron spectroscopy as the system was tuned to its quantum critical point by an applied magnetic field. The electronic mode softening expected for a quantum phase transition was forestalled by hyperfine coupling to the nuclear spins. We found that interactions with the nuclear spin bath controlled the length scale over which the excitations could be entangled. This generic result places a limit on our ability to observe intrinsic electronic quantum criticality. PMID- 15831753 TI - Atomic-scale visualization of inertial dynamics. AB - The motion of atoms on interatomic potential energy surfaces is fundamental to the dynamics of liquids and solids. An accelerator-based source of femtosecond x ray pulses allowed us to follow directly atomic displacements on an optically modified energy landscape, leading eventually to the transition from crystalline solid to disordered liquid. We show that, to first order in time, the dynamics are inertial, and we place constraints on the shape and curvature of the transition-state potential energy surface. Our measurements point toward analogies between this nonequilibrium phase transition and the short-time dynamics intrinsic to equilibrium liquids. PMID- 15831754 TI - A convergent enantioselective route to structurally diverse 6-deoxytetracycline antibiotics. AB - Complex antibiotics based on natural products are almost invariably prepared by semisynthesis, or chemical transformation of the isolated natural products. This approach greatly limits the range of accessible structures that might be studied as new antibiotic candidates. Here we report a short and enantioselective synthetic route to a diverse range of 6-deoxytetracycline antibiotics. The common feature of this class is a scaffold of four linearly fused rings, labeled A through D. We targeted not a single compound but a group of structures with the D ring as a site of structural variability. A late-stage, diastereoselective C-ring construction was used to couple structurally varied D-ring precursors with an AB precursor containing much of the essential functionality for binding to the bacterial ribosome. Five derivatives were synthesized from benzoic acid in yields ranging from 5 to 7% over 14 to 15 steps, and a sixth, (-)-doxycycline, was synthesized in 8.3% yield over 18 steps. PMID- 15831755 TI - Hypoxia, global warming, and terrestrial late Permian extinctions. AB - A catastrophic extinction occurred at the end of the Permian Period. However, baseline extinction rates appear to have been elevated even before the final catastrophe, suggesting sustained environmental degradation. For terrestrial vertebrates during the Late Permian, the combination of a drop in atmospheric oxygen plus climate warming would have induced hypoxic stress and consequently compressed altitudinal ranges to near sea level. Our simulations suggest that the magnitude of altitudinal compression would have forced extinctions by reducing habitat diversity, fragmenting and isolating populations, and inducing a species area effect. It also might have delayed ecosystem recovery after the mass extinction. PMID- 15831756 TI - Open-system coral ages reveal persistent suborbital sea-level cycles. AB - Sea level is a sensitive index of global climate that has been linked to Earth's orbital variations, with a minimum periodicity of about 21,000 years. Although there is ample evidence for climate oscillations that are too frequent to be explained by orbital forcing, suborbital-frequency sea-level change has been difficult to resolve, primarily because of problems with uranium/thorium coral dating. Here we use a new approach that corrects coral ages for the frequently observed open-system behavior of uranium-series nuclides, substantially improving the resolution of sea-level reconstruction. This curve reveals persistent sea level oscillations that are too frequent to be explained exclusively by orbital forcing. PMID- 15831757 TI - Fragmentation and flow regulation of the world's large river systems. AB - A global overview of dam-based impacts on large river systems shows that over half (172 out of 292) are affected by dams, including the eight most biogeographically diverse. Dam-impacted catchments experience higher irrigation pressure and about 25 times more economic activity per unit of water than do unaffected catchments. In view of projected changes in climate and water resource use, these findings can be used to identify ecological risks associated with further impacts on large river systems. PMID- 15831758 TI - A homolog of Drosophila grainy head is essential for epidermal integrity in mice. AB - The Drosophila cuticle is essential for maintaining the surface barrier defenses of the fly. Integral to cuticle resilience is the transcription factor grainy head, which regulates production of the enzyme required for covalent cross linking of the cuticular structural components. We report that formation and maintenance of the epidermal barrier in mice are dependent on a mammalian homolog of grainy head, Grainy head-like 3. Mice lacking this factor display defective skin barrier function and deficient wound repair, accompanied by reduced expression of transglutaminase 1, the key enzyme involved in cross-linking the structural components of the superficial epidermis. These findings suggest that the functional mechanisms involving protein cross-linking that maintain the epidermal barrier and induce tissue repair are conserved across 700 million years of evolution. PMID- 15831759 TI - Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro. AB - Kai proteins globally regulate circadian gene expression of cyanobacteria. The KaiC phosphorylation cycle, which persists even without transcription or translation, is assumed to be a basic timing process of the circadian clock. We have reconstituted the self-sustainable oscillation of KaiC phosphorylation in vitro by incubating KaiC with KaiA, KaiB, and adenosine triphosphate. The period of the in vitro oscillation was stable despite temperature change (temperature compensation), and the circadian periods observed in vivo in KaiC mutant strains were consistent with those measured in vitro. The enigma of the circadian clock can now be studied in vitro by examining the interactions between three Kai proteins. PMID- 15831760 TI - Representation of visual gravitational motion in the human vestibular cortex. AB - How do we perceive the visual motion of objects that are accelerated by gravity? We propose that, because vision is poorly sensitive to accelerations, an internal model that calculates the effects of gravity is derived from graviceptive information, is stored in the vestibular cortex, and is activated by visual motion that appears to be coherent with natural gravity. The acceleration of visual targets was manipulated while brain activity was measured using functional magnetic resonance imaging. In agreement with the internal model hypothesis, we found that the vestibular network was selectively engaged when acceleration was consistent with natural gravity. These findings demonstrate that predictive mechanisms of physical laws of motion are represented in the human brain. PMID- 15831761 TI - Keys to Independence in the U.K. and Ireland. AB - Funding bodies are coming to the aid of young scientists with a variety of programs aimed at helping them become independent investigators. PMID- 15831762 TI - Views From the Trenches. AB - Making the transition from postdoc to independent investigator is tough. Scientists who received prestigious fellowships or grants relate their experiences-and offer some advice. PMID- 15831763 TI - Phasic pulmonary stretch receptor feedback modulates both eupnea and gasping in an in situ rat preparation. AB - The perfused in situ juvenile rat preparation produces patterns of phrenic discharge comparable to eupnea and gasping in vivo. These ventilatory patterns differ in multiple aspects, including most prominently the rate of rise of inspiratory activity. Although we have recently demonstrated that both eupnea and gasping are similarly modulated by a Hering-Breuer expiratory-promoting reflex to tonic pulmonary stretch, it has generally been assumed that gasping was unresponsive to afferent stimuli from pulmonary stretch receptors. In the present study, we recorded eupneic and gasplike efferent activity of the phrenic nerve in the in situ juvenile rat perfused brain stem preparation, with and without phrenic-triggered phasic pulmonary inflation. We tested the hypothesis that phasic pulmonary inflation produces reflex responses in situ akin to those in vivo and that both eupnea and gasping are similarly modulated by phasic pulmonary stretch. In eupnea, we found that phasic pulmonary inflation decreases inspiratory burst duration and the period of expiration, thus increasing burst frequency of the phrenic neurogram. Phasic pulmonary inflation also decreases the duration of expiration and increases the burst frequency during gasping. Bilateral vagotomy eliminated these changes. We conclude that the neural substrate mediating the Hering-Breuer reflex is retained in the in situ preparation and that the brain stem circuitry generating the respiratory patterns respond to phasic activation of pulmonary stretch receptors in both eupnea and gasping. These findings support the homology of eupneic phrenic discharge patterns in the reduced in situ preparation and eupnea in vivo and disprove the common supposition that gasping is insensitive to vagal afferent feedback from pulmonary stretch receptor mechanisms. PMID- 15831764 TI - Angiotensin depolarizes parvocellular neurons in paraventricular nucleus through modulation of putative nonselective cationic and potassium conductances. AB - Neurosecretory parvocellular neurons in the hypothalamic paraventricular nucleus (PVN) exercise considerable influence over the adenohypophysis and thus play a critical role in neuroendocrine regulation. ANG II has been demonstrated to act as a neurotransmitter in PVN, exerting significant impact on neuronal excitability and also influencing corticotrophin-releasing hormone secretion from the median eminence and, therefore, release of ACTH from the pituitary. We have used whole cell patch-clamp techniques in hypothalamic slices to examine the effects of ANG II on the excitability of neurosecretory parvocellular neurons. ANG II application resulted in a dose-dependent depolarization of neurosecretory neurons, a response that was maintained in tetrodotoxin (TTX), suggesting a direct mechanism of action. The depolarizing actions of this peptide were abolished by losartan, demonstrating these effects are AT(1) receptor mediated. Voltage-clamp analysis using slow voltage ramps revealed that ANG II activates a voltage-independent conductance with a reversal potential of -37.8 +/- 3.8 mV, suggesting ANG II effects on a nonselective cationic current. Further, a sustained potassium current characteristic of I(K) was significantly reduced (29.1 +/- 4.7%) by ANG II. These studies identify multiple postsynaptic modulatory sites through which ANG II can influence the excitability of neurosecretory parvocellular PVN neurons and, as a consequence of such actions, control hormonal secretion from the anterior pituitary. PMID- 15831765 TI - Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology. AB - Leptin is a multifunctional hormone, produced predominantly in adipocytes. It regulates energy balance through its impact on appetite and fat metabolism, and its concentration indicates the size of body fat reserves. Leptin also plays a vital role in stretch-induced surfactant production during alveolar development in the fetus. The structure, expression pattern, and role of leptin have not previously been explored in marine mammals. Phocid seals undergo cyclical changes in body composition as a result of prolonged fasting and intensive foraging bouts and experience rapid, dramatic, and repeated changes in lung volume during diving. Here, we report the tissue-specific expression pattern of leptin in these animals. This is the first demonstration of leptin expression in the lung tissue of a mature mammal, in addition to its expression in the blubber and bone marrow, in common with other animals. We propose a role for leptin in seal pulmonary surfactant production, in addition to its likely role in long-term energy balance. We identify substitutions in the phocine leptin sequence in regions normally highly conserved between widely distinct vertebrate groups, and, using a purified seal leptin antiserum, we confirm the presence of the leptin protein in gray seal lung and serum fractions. Finally, we report the substantial inadequacies of using heterologous antibodies to measure leptin in unextracted gray seal serum. PMID- 15831767 TI - Gastrointestinal tract innervation of the mouse: afferent regeneration and meal patterning after vagotomy. AB - Mice, with the variety of genotypes they provide, should be particularly useful for studies of growth factors and gene products in regeneration of autonomic pathways such as the vagus nerve. To provide a foundation for examinations of mouse vagal reorganization, two experiments assessed the rate, extent, and accuracy of afferent reinnervation of the stomach after vagotomy and related these patterns to feeding behavior. In experiment 1, the pattern of afferent regrowth into the gut after unilateral truncal vagotomy was characterized by labeling of these afferents with wheat germ agglutinin-horseradish peroxidase and Micro-Ruby. Regenerating neurites had reached and, in some cases, already reinnervated the stomach by 4 wk after axotomy. By 8 wk, regrowth was more extensive, and many fibers had redifferentiated terminals in the smooth muscle. By 16 wk, vagal projections had reached or exceeded normal density in the corpus, density in the forestomach was still reduced, and regrowth in the antrum was minimal. At all time points, not only appropriate terminals, but also growth cones and aberrant endings, were observed. In experiment 2, meal patterns of vagotomized mice were evaluated using a solid diet over the period of regeneration; cholecystokinin suppression of a liquid meal after unilateral and bilateral truncal vagotomies was also evaluated. Unilaterally, as well as bilaterally, vagotomized animals ate smaller and more frequent meals. These disturbed patterns became more pronounced in the first 8 wk after vagotomy, during regeneration. Cholecystokinin inhibition of intake was attenuated by bilateral, but not unilateral, vagotomy. Overall, the spatial and temporal patterns of structural and functional changes observed during regeneration verify that the mouse provides a useful preparation for examining the control of vagal plasticity. PMID- 15831766 TI - Corticotropin-releasing factor receptor subtypes mediating nutritional suppression of estrous behavior in Syrian hamsters. AB - Caloric deprivation inhibits reproduction, including copulatory behaviors, in female mammals. Decreases in metabolic fuel availability are detected in the hindbrain, and this information is relayed to the forebrain circuits controlling estrous behavior by neuropeptide Y (NPY) projections. In the forebrain, the nutritional inhibition of estrous behavior appears to be mediated by corticotropin-releasing factor (CRF) or urocortin-signaling systems. Intracerebroventricular (ICV) infusion of the CRF antagonist, astressin, prevents the suppression of lordosis by food deprivation and by NPY treatment in Syrian hamsters. These experiments sought to determine which CRF receptor type(s) is involved. ICV infusion of the CRF receptor subtype CRFR2-selective agonists urocortin 2 and 3 (UCN2, UCN3) inhibited sexual receptivity in hormone-primed, ovariectomized hamsters. Furthermore, the CRFR2-selective antagonist, astressin 2B, prevented the inhibition of estrous behavior by UCN2 and by NPY, consistent with a role for CRFR2. On the other hand, astressin 2B did not prevent the inhibition of behavior induced by 48-h food deprivation or ICV administration of CRF, a mixed CRFR1 and CRFR2 agonist, suggesting that activation of CRFR1 signaling is sufficient to inhibit sexual receptivity in hamsters. Although administration of CRFR1-selective antagonists (NBI-27914 and CP-154,526) failed to reverse the inhibition of receptivity by CRF treatment, we could not confirm their biological effectiveness in hamsters. The most parsimonious interpretation of these findings is that, although NPY inhibits estrous behavior via downstream CRFR2 signaling, food deprivation may exert its inhibition via both CRFR1 and CRFR2 and that redundant neuropeptide systems may be involved. PMID- 15831768 TI - Localized vs. systemic inflammation in guinea pigs: a role for prostaglandins at distinct points of the fever induction pathways? AB - In guinea pigs, dose-dependent febrile responses were induced by injection of a high (100 microg/kg) or a low (10 microg/kg) dose of bacterial lipopolysaccharide (LPS) into artificial subcutaneously implanted Teflon chambers. Both LPS doses further induced a pronounced formation of prostaglandin E(2) (PGE(2)) at the site of localized subcutaneous inflammation. Administration of diclofenac, a nonselective cyclooxygenase (COX) inhibitor, at different doses (5, 50, 500, or 5,000 microg/kg) attenuated or abrogated LPS-induced fever and inhibited LPS induced local PGE(2) formation (5 or 500 microg/kg diclofenac). Even the lowest dose of diclofenac (5 microg/kg) attenuated fever in response to 10 microg/kg LPS, but only when administered directly into the subcutaneous chamber, and not into the site contralateral to the chamber. This observation indicated that a localized formation of PGE(2) at the site of inflammation mediated a portion of the febrile response, which was induced by injection of 10 microg/kg LPS into the subcutaneous chamber. Further support for this hypothesis derived from the observation that we failed to detect elevated amounts of COX-2 mRNA in the brain of guinea pigs injected subcutaneously with 10 microg/kg LPS, whereas subcutaneous injections of 100 microg/kg LPS, as well as systemic injections of LPS (intra-arterial or intraperitoneal routes), readily caused expression of the COX-2 gene in the guinea pig brain, as demonstrated by in situ hybridization. Therefore, fever in response to subcutaneous injection of 10 microg/kg LPS may, in part, have been evoked by a neural, rather than a humoral, pathway from the local site of inflammation to the brain. PMID- 15831769 TI - Modulation of apoptotic pathways in intestinal mucosa during hibernation. AB - Mammalian hibernation is associated with several events that can affect programmed cell death (apoptosis) in nonhibernators, including marked changes in blood flow, extended fasting, and oxidative stress. However, the effect of hibernation on apoptosis is poorly understood. Here, we investigated apoptosis and expression of proteins involved in apoptotic pathways in intestinal mucosa of summer and hibernating ground squirrels. We used terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) to identify possible apoptotic enterocytes in small intestine of summer squirrels and hibernating squirrels throughout the winter. Nuclear TUNEL staining increased as hibernation progressed, but the staining pattern was diffuse and not accompanied by chromatin condensation or apoptotic bodies. Electrophoresis of mucosal DNA revealed no ladders typical of apoptosis. Nuclear levels of proapoptotic p53 protein were fourfold less in hibernators compared with summer squirrels. A 12-fold increase in anti-apoptotic Bcl-x(L) compared with a 2-fold increase in proapoptotic Bax suggested a balance in favor of antiapoptotic signaling in hibernators. There was no change in Bcl-2 protein expression but phospho-Bcl-2 increased in mucosa of hibernators. Hibernation had minimal effects on expression of active caspase-8 or -9, whereas caspase-3-specific activity was lower in hibernators during an interbout arousal compared with summer squirrels. Expression of the prosurvival protein Akt increased 20-fold during hibernation, but phospho-Akt was not altered. These data provide evidence for enhanced expression of antiapoptotic proteins during hibernation that may promote enterocyte survival in a pro oxidative, proapoptotic environment. PMID- 15831770 TI - Inhibitory effects on intake of cholecystokinin-8 and cholecystokinin-33 in rats with hepatic proper or common hepatic branch vagal innervation. AB - The relative potencies of cholecystokinin (CCK)-8 and CCK-33 for decreasing meal size depend on the route of administration. Inhibitory potencies are equal after intraperitoneal administration, but CCK-33 is significantly more potent after intraportal administration. This suggests that CCK-33 is a more effective stimulant of hepatic afferent vagal nerves than is CCK-8. To investigate this possibility, we administered both peptides intraperitoneally in rats with abdominal vagotomies that spared only the hepatic proper vagal nerves (H) and in rats with abdominal vagotomies that spared the common hepatic branch that contains the fibers of the hepatic proper and gastroduodenal nerves (HGD). The vagal afferent innervation in H and HGD rats was verified with a wheat germ agglutinin-horseradish tracer strategy. Intraperitoneal administration of CCK-33 decreased 30-min intake of 10% sucrose in H rats as much as in sham rats, but CCK 8 decreased intake significantly less in H rats than in sham rats. The larger inhibitory effect of CCK-33 than of CCK-8 in H rats is consistent with the hypothesis that CCK-33 is a more effective stimulant of the hepatic proper vagal afferent nerves than CCK-8. In contrast to the results in H rats, the inhibitory potencies of both peptides were significantly and equivalently reduced in HGD rats compared with sham rats. This suggests that there is an inhibitory interaction between the stimulation of the gastroduodenal and hepatic proper afferent fibers by CCK-33. PMID- 15831771 TI - Clinical pharmacokinetics of etanercept: a fully humanized soluble recombinant tumor necrosis factor receptor fusion protein. AB - Etanercept, a fully humanized soluble recombinant tumor necrosis factor receptor fusion protein, is an approved treatment for rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, and psoriasis. Etanercept is absorbed slowly from the site of subcutaneous injection, with time to peak concentration at approximately 48 to 60 hours, and is cleared slowly from the body with a t(1/2) of 70 to 100 hours. The absolute bioavailability of etanercept was 58% in healthy subjects following subcutaneous administration. The 25-mg twice-weekly dosage regimen generates systemic exposures comparable to 50 mg once weekly, as predicted by pharmacokinetic modeling and simulation and later confirmed by clinical studies. The pharmacokinetics of etanercept in patients with rheumatoid arthritis are comparable to those in healthy individuals and patients with ankylosing spondylitis, congestive heart failure, and psoriasis. In children with polyarticular-course juvenile rheumatoid arthritis, after subcutaneous doses of 0.4 mg/kg twice weekly, the clearance of etanercept may be slightly reduced in children aged 4 to 8 years. Pharmacokinetic simulation predicts that a dose of 0.8 mg/kg once weekly generates comparable systemic exposure as 0.4 mg/kg twice weekly. No requirement for etanercept dosage adjustment is needed when etanercept is coadministered with warfarin, digoxin, or methotrexate. PMID- 15831772 TI - Validation of the human ozone challenge model as a tool for assessing anti inflammatory drugs in early development. AB - This study aimed to test the utility of the ozone challenge model for profiling novel compounds designed to reduce airway inflammation. The authors used a randomized, double-dummy, double-blind, placebo-controlled 3-period crossover design alternating single orally inhaled doses of fluticasone propionate (inhaled corticosteroids, 2 mg), oral prednisolone (oral corticosteroids, 50 mg), or matched placebo. At a 2-week interval, 18 healthy ozone responders (>10% increase in sputum neutrophils) underwent a 3-hour ozone (250 ppb)/intermittent exercise challenge starting 1 hour after drug treatment. Airway inflammation was assessed at 2 hours (breath condensate) and 3 hours (induced sputum) after ozone challenge. Compared to placebo, pretreatment with inhaled corticosteroids or oral corticosteroids resulted in a significant reduction (mean [95% confidence interval]) of sputum neutrophils by 62% (35%, 77%) and 64% (39%, 79%) and of sputum supernatant myeloperoxidase by 55% (41%, 66%) and 42% (25%, 56%), respectively. The authors conclude that an optimized ozone challenge model (including ozone responders and ensuring adequate drug levels during exposure) may be useful for testing novel anti-inflammatory compounds in early development. PMID- 15831773 TI - Individual and combined effects of peroxisome proliferator-activated receptor and {gamma} agonists, fenofibrate and rosiglitazone, on biomarkers of lipid and glucose metabolism in healthy nondiabetic volunteers. AB - This open-label, randomized, placebo-controlled, incomplete-block, 3-period crossover pilot study investigated the effects of peroxisome proliferator activated receptor alpha- and gamma-agonists on biomarkers of lipid and glucose metabolism in 12 nondiabetic subjects. Plasma samples were collected before and after each 14-day treatment with placebo, fenofibrate (201 mg/d), rosiglitazone (4 mg twice daily), and combined fenofibrate (201 mg/d) plus rosiglitazone (4 mg twice daily). Except for triglycerides (P < .042) and free fatty acids (P < .074), no significant interaction was demonstrated between fenofibrate and rosiglitazone; thus, the effect due to each drug alone was evaluated (presence/absence of drug). Fenofibrate significantly (P < .050) increased lipoprotein lipase activity (35%) and decreased apolipoproteins B (13%) and C-III (20%). Rosiglitazone significantly (P < .050) decreased fasting glucose (7.3%) and increased apolipoprotein C-III (19%) and adiponectin (137%). Fenofibrate and rosiglitazone also produced effects on triglycerides and free fatty acids, but it was not possible to determine if these effects were synergistic in nature. PMID- 15831774 TI - Blood concentrations of everolimus are markedly increased by ketoconazole. AB - The authors sought to quantify the influence of the CYP3A and P-glycoprotein inhibitor ketoconazole on the pharmacokinetics of everolimus in healthy subjects. This was a 2-period, single-sequence, crossover study in 12 healthy subjects. In period 1, subjects received the reference treatment of a single 2-mg dose of everolimus. In period 2, they received the test treatment of ketoconazole 200 mg twice daily for a total of 8 days and a single dose of everolimus coadministered on the fourth day of ketoconazole therapy. The test/reference ratio and 90% confidence interval were derived for everolimus maximum concentration and area under the curve. During ketoconazole coadministration, everolimus maximum concentration increased 3.9-fold (90% confidence interval, 3.4-4.6) from 15 +/- 4 ng/mL to 59 +/- 13 ng/mL. Everolimus area under the curve increased 15.0-fold (90% confidence interval, 13.6-16.6) from 90 +/- 23 ng*h/mL to 1324 +/- 232 ng*h/mL. Everolimus half-life was prolonged by 1.9-fold from 30 +/- 4 hours to 56 +/- 5 hours. Everolimus did not appear to alter ketoconazole predose concentrations. Given the magnitude of this drug interaction, use of ketoconazole should be avoided if possible in everolimus-treated patients. PMID- 15831775 TI - Steady-state pharmacokinetics of galantamine are not affected by addition of memantine in healthy subjects. AB - To evaluate the effect of multiple doses of memantine on the pharmacokinetics of galantamine and to assess the safety and tolerability of galantamine with adjunctive memantine treatment, an open-label, single-center, drug interaction study was conducted in 16 healthy adults. Subjects received an 8-mg dose of galantamine extended release once daily during week 1 and a 16-mg dose of galantamine extended release once daily during week 2. During weeks 3 and 4, they received a 16-mg dose of galantamine extended release once daily and a 10-mg dose of memantine twice daily, except on days 1 and 2 of week 3, when memantine was given as 10 mg once daily. The pharmacokinetic profile and parameters of galantamine at steady state were similar after administration of a 16-mg dose of galantamine once daily alone and after administration with a 10-mg dose of memantine twice daily. Galantamine 16 mg once daily with adjunctive memantine 10 mg twice daily was well tolerated and safe in healthy subjects. PMID- 15831776 TI - Kinetics and dynamics of intravenous adinazolam, N-desmethyl adinazolam, and alprazolam in healthy volunteers. AB - The pharmacokinetics and pharmacodynamics of adinazolam mesylate (10 mg), N desmethyl adinazolam mesylate (NDMAD, 10 mg), and alprazolam (1 mg) were investigated in 9 healthy male subjects in a randomized, blinded, single-dose, 4 way crossover study. All drugs were intravenously infused over 30 minutes. Plasma adinazolam, NDMAD, and alprazolam concentrations, electroencephalographic (EEG) activity in the beta (12-30 Hz) range, performance on the Digit Symbol Substitution Test (DSST), and subjective measures of mood and sedation were monitored for 12 to 24 hours. Mean pharmacokinetic parameters for adinazolam, NDMAD, and alprazolam, respectively, were as follows: volume of distribution (L), 106, 100, and 77; elimination half-life (hours), 2.9, 2.8, and 14.6; and clearance (mL/min), 444, 321, and 84. More than 80% of the total infused adinazolam dose was converted to systemically appearing NDMAD. All 3 benzodiazepine agonists significantly increased beta EEG activity, with alprazolam showing the strongest agonist activity and adinazolam showing the weakest activity. Alprazolam and NDMAD significantly decreased DSST performance, whereas adinazolam had no effect relative to placebo. Adinazolam, NDMAD, and alprazolam all produced significant observer-rated sedation. Plots of EEG effect versus plasma alprazolam concentration demonstrated counterclockwise hysteresis, consistent with an effect site delay. This was incorporated into a kinetic dynamic model in which hypothetical effect site concentration was related to pharmacodynamic EEG effect via the sigmoid E(max) model, yielding an effect site equilibration half-life of 4.8 minutes. The exponential effect model described NDMAD pharmacokinetics and EEG pharmacodynamics. The relation of both alprazolam and NDMAD plasma concentrations to DSST performance could be described by a modified exponential model. Pharmacokinetic-dynamic modeling was not possible for adinazolam, as the data did not conform to any known concentration-effect model. Collectively, these results indicate that the benzodiazepine-like effects occurring after adinazolam administration are mediated by mainly NDMAD. PMID- 15831777 TI - Tolerability, gut effects, and pharmacokinetics of methylnaltrexone following repeated intravenous administration in humans. AB - Previous studies have shown that a single dose of methylnaltrexone, a unique peripheral opioid antagonist, reverses opioid-induced gut hypomotility in humans. Because repeated drug doses are likely to be needed to treat patients with opioid induced or postsurgical bowel dysfunction, the authors have now examined the safety, pharmacological activity, and pharmacokinetics of a multiple-dose regimen of methylnaltrexone, administered as 12 consecutive intravenous doses (0.3 mg/kg every 6 hours) in 12 healthy subjects. Steady state was achieved rapidly, and after repeated dosing for 3 days, methylnaltrexone decreased oral-cecal transit time from a pretreatment baseline value of 101.3 +/- 29.4 min (mean +/- SD) to 82.5 +/- 20.7 min. Maximum observed plasma concentrations, measured 5 minutes postdose, were 538 +/- 237 and 675 +/- 180 ng/mL after doses 1 and 2, respectively. Based on 6-hour sampling periods, the plasma half-life, 2.5 +/- 0.5 and 2.9 +/- 0.9 hours following the 1st and 12th doses, respectively, was unchanged at steady state. There was essentially no accumulation of methylnaltrexone, based on the ratio of AUC values after doses 12 and 1. This study showed that repeated administration of intravenous methylnaltrexone is well tolerated in humans, with no significant adverse events or changes in opioid subjective ratings and no clinically noteworthy alterations in pharmacokinetics. The observation of a significant reduction in the gut transit time after repeated administration of methylnaltrexone to these opioid-naive volunteers suggests that endogenous opioids modulate human gut motility. PMID- 15831778 TI - Clinical pharmacology and pharmacokinetics of once-daily hydromorphone hydrochloride extended-release capsules. AB - Hydromorphone hydrochloride extended release (HHER) is a multiparticulate melt extrusion pellet capsule formulation administered q24h. Study 1 investigated the bioavailability of 24-mg HHER fed, as well as 24-mg and 12-mg HHER and 8-mg hydromorphone hydrochloride immediate-release (HHIR) tablets fasting. No clinically significant food effect was observed on hydromorphone C(max) or AUC for the 24-mg HHER, and dose proportionality (AUC) was demonstrated between 12- and 24-mg HHER. Study 2 demonstrated dose strength proportionality for 3 x 12-mg HHER versus 1 x 32-mg HHER. Study 3 evaluated 12-mg HHER q24h versus 3-mg HHIR q6h at steady state. HHER produced relatively constant steady-state concentrations over 24 hours. HHER and HHIR were equivalent for AUC(ss). C(ssmax) was 26% lower for HHER than HHIR, C(ssmin) was 43% higher for HHER, and peak-to trough fluctuation was 126% for HHER versus 328% for HHIR, which are ideal attributes of a once-daily oral extended-release dosage form. HHER administration resulted in fewer adverse events than HHIR in study 3. PMID- 15831779 TI - Pharmacokinetic profile of the oral direct thrombin inhibitor dabigatran etexilate in healthy volunteers and patients undergoing total hip replacement. AB - Dabigatran etexilate is an oral low-molecular-weight direct thrombin inhibitor. Following oral administration, dabigatran etexilate is rapidly converted to its active form, dabigatran. The authors investigated the absorption, distribution, and elimination of a single 150-mg dose capsule formulation of dabigatran etexilate in healthy volunteers and patients undergoing total hip replacement. In an open-label, 3-way crossover study, dabigatran etexilate was administered to 18 male volunteers in the fasted state, after administration of food and with coadministration of the proton pump inhibitor, pantoprazole. In a subsequent multicenter, open-label study, 59 patients received a single dose of dabigatran etexilate, administered 1 to 3 hours following total hip replacement. In healthy volunteers, food had no effect on the extent of absorption of dabigatran etexilate, although there was reduced interindividual variability for dabigatran maximum plasma concentration and AUC(0-infinity). A decrease in the mean dabigatran AUC(0-infinity) (904 to 705 ng*h/mL) occurred with coadministration of pantoprazole. In patients undergoing total hip replacement, immediate onset of absorption was seen with the maximum plasma concentration of dabigatran occurring after 6 hours. The AUC(0-24) of dabigatran was 88% of the steady-state AUC using a preliminary tablet formulation and 106% of that seen in the healthy volunteer study. Compared with healthy volunteers, the postoperative profile was flattened with delayed peak concentrations. In summary, administration of the dabigatran etexilate capsule with food has no effect on the extent of dabigatran absorption, with a moderate decrease when coadministered with pantoprazole. Adequate plasma concentrations of dabigatran were seen with early postoperative administration of the dabigatran etexilate capsule. These pharmacokinetic characteristics confirm the suitability of this oral solid dosage form for use in future clinical trials. PMID- 15831780 TI - Tissue penetration of cefpodoxime and cefixime in healthy subjects. AB - Microdialysis is a technique that allows the measurement of free antibiotic concentrations in different tissues, which are responsible for the antibacterial activity at the infection site. In an open, randomized, 2-way crossover study in healthy volunteers, the muscle penetration of orally administered cefpodoxime (400 mg) and cefixime (400 mg) was compared using microdialysis. The results show that the total plasma concentration-time profiles of each antibiotic were similar; the area under the curve for cefpodoxime was 22.4 +/- 8.7 versus 25.6 +/ 8.5 mg/L*h for cefixime. However, tissue penetration was twice as high for cefpodoxime (area under the curve 15.4 +/- 5.1 mg/L*h) as for cefixime (area under the curve 7.3 mg/L*h). This degree of tissue distribution is consistent with their protein binding of 21% for cefpodoxime and 65% for cefixime. After equilibration, the unbound tissue concentrations of both antibiotics were similar to their unbound plasma concentrations. Pharmacokinetic modeling was applied to describe the pharmacokinetic profiles in plasma and muscle. The study demonstrates that cefpodoxime shows greater tissue penetration than cefixime. PMID- 15831781 TI - Pharmacokinetics of an oral drug (acetaminophen) administered at various times in relation to subcutaneous injection of exenatide (exendin-4) in healthy subjects. AB - Exenatide is an incretin mimetic with potential glucoregulatory activity in type 2 diabetes. This randomized, single-blind, placebo-controlled 6-way crossover study assessed exenatide's effect on acetaminophen pharmacokinetics. Of 40 randomized healthy subjects, 39 completed the study. On the placebo day, acetaminophen (1000 mg) was ingested and placebo injected subcutaneously at 0 hours. On exenatide days, acetaminophen was ingested at -1, 0, +1, +2, and +4 hours, relative to the 10 mug exenatide injected subcutaneously at 0 hours. With exenatide injection, mean plasma acetaminophen AUC(0-12 h) values were reduced by 11% to 24% (vs placebo). Peak plasma acetaminophen concentrations were similar for the -1-hour and placebo groups and reduced by 37% to 56% at other times. The most frequent adverse events were generally mild to moderate nausea and vomiting. Exenatide treatment concurrent with or preceding acetaminophen ingestion slowed acetaminophen absorption but had minimal effect on the extent of absorption. PMID- 15831782 TI - Single- and multiple-dose pharmacokinetics of levovirin valinate hydrochloride (R1518) in healthy volunteers. AB - R1518 is a valine ester prodrug of levovirin as an investigational new drug for the treatment of hepatitis C virus. Two phase 1, single- and multiple-dose studies were conducted to investigate the pharmacokinetics of R1518 in healthy volunteers. After oral dosing, R1518 was rapidly and exclusively converted to levovirin. Levovirin plasma concentrations peaked at 2 hours, with T(1/2) ranging from 6 to 8 hours. The T(1/2) of R1518 was less than 1 hour, with relative exposures (R1518/levovirin) less than 6%. A high-fat meal did not affect the pharmacokinetics. The female groups in both studies had higher plasma levels than males did due to age and renal function difference. An accumulation ratio of 1.3 to 1.5 was observed with the twice-daily regimen. About 75% to 90% of the levovirin equivalent dose was recovered in urine. Increase in exposure was slightly disproportionate to increase in dose. Significantly improved oral absorption of levovirin was achieved following administration of R1518. PMID- 15831783 TI - Evaluation of safety and pharmacokinetics of consecutive multiple-day dosing of palonosetron in healthy subjects. AB - This study evaluated the safety and pharmacokinetics of consecutive multiple-day dosing of palonosetron. Sixteen healthy subjects received an intravenous bolus dose of palonosetron 0.25 mg (n = 12) or placebo (n = 4) daily for 3 consecutive days. Safety was evaluated throughout the study. Serial plasma samples were collected on days 1 and 3 for pharmacokinetic determinations. Three days of dosing with palonosetron 0.25 mg was safe and well tolerated. There were no clinically significant changes from baseline in laboratory values, vital signs, physical examinations, or electrocardiogram intervals. Plasma palonosetron concentrations declined in a biphasic manner, measurable up to 168 hours after dosing on day 3. Mean terminal phase elimination half-life after day 3 dosing was 42.8 hours. The 2.1-fold accumulation of palonosetron in plasma following 3 daily doses was predictable based on elimination half-life of approximately 40 hours, and the maximum plasma concentration remained below the maximum plasma concentration previously observed after a single, well-tolerated 0.75 mg intravenous bolus dose of palonosetron. PMID- 15831784 TI - Bioavailability of azacitidine subcutaneous versus intravenous in patients with the myelodysplastic syndromes. AB - The primary objectives of this study were to characterize the absolute bioavailability of azacitidine after subcutaneous (SC) administration and to compare the single-dose pharmacokinetics of azacitidine following SC and intravenous (IV) administration. Six patients with myelodysplastic syndromes were randomly assigned according to a crossover design to treatment A, consisting of azacitidine administered as a single 75-mg/m(2) SC dose, or treatment B, consisting of azacitidine administered as a single 75-mg/m(2) IV infusion dose over 10 minutes. A minimum of 7 days and a maximum of 28 days were permitted between treatments. The study demonstrated good bioavailability of a SC azacitidine dose compared to an IV infusion treatment. The exposure profiles following SC drug administration illustrate measurable azacitidine levels with bioavailability (AUC) values within 89% of those measured following IV administration (range, 70%-112%). The median IV half-life was 0.36 +/- 0.02 hours compared to 0.69 +/- 0.14 hours for SC administration. Regardless of the route of administration, a single dose of azacitidine, 75 mg/m(2), was generally well tolerated. PMID- 15831785 TI - Single copy shRNA configuration for ubiquitous gene knockdown in mice. AB - RNA interference through the expression of small hairpin RNA (shRNA) molecules has become a very promising tool in reverse mouse genetics as it may allow inexpensive and rapid gene function analysis in vivo. However, the prerequisites for ubiquitous and reproducible shRNA expression are not well defined. Here we show that a single copy shRNA-transgene can mediate body-wide gene silencing in mice when inserted in a defined locus of the genome. The most commonly used promoters for shRNA expression, H1 and U6, showed a comparably broad activity in this configuration. Taken together, the results define a novel approach for efficient interference with expression of defined genes in vivo. Moreover, we provide a rapid strategy for the production of gene knockdown mice combining recombinase mediated cassette exchange and tetraploid blastocyst complementation approaches. PMID- 15831786 TI - Detecting DNA-binding helix-turn-helix structural motifs using sequence and structure information. AB - In this work, we analyse the potential for using structural knowledge to improve the detection of the DNA-binding helix-turn-helix (HTH) motif from sequence. Starting from a set of DNA-binding protein structures that include a functional HTH motif and have no apparent sequence similarity to each other, two different libraries of hidden Markov models (HMMs) were built. One library included sequence models of whole DNA-binding domains, which incorporate the HTH motif, the second library included shorter models of 'partial' domains, representing only the fraction of the domain that corresponds to the functionally relevant HTH motif itself. The libraries were scanned against a dataset of protein sequences, some containing the HTH motifs, others not. HMM predictions were compared with the results obtained from a previously published structure-based method and subsequently combined with it. The combined method proved more effective than either of the single-featured approaches, showing that information carried by motif sequences and motif structures are to some extent complementary and can successfully be used together for the detection of DNA-binding HTHs in proteins of unknown function. PMID- 15831787 TI - Ribonucleases J1 and J2: two novel endoribonucleases in B.subtilis with functional homology to E.coli RNase E. AB - Many prokaryotic organisms lack an equivalent of RNase E, which plays a key role in mRNA degradation in Escherichia coli. In this paper, we report the purification and identification by mass spectrometry in Bacillus subtilis of two paralogous endoribonucleases, here named RNases J1 and J2, which share functional homologies with RNase E but no sequence similarity. Both enzymes are able to cleave the B.subtilis thrS leader at a site that can also be cleaved by E.coli RNase E. We have previously shown that cleavage at this site increases the stability of the downstream messenger. Moreover, RNases J1/J2 are sensitive to the 5' phosphorylation state of the substrate in a site-specific manner. Orthologues of RNases J1/J2, which belong to the metallo-beta-lactamase family, are evolutionarily conserved in many prokaryotic organisms, representing a new family of endoribonucleases. RNases J1/J2 appear to be implicated in regulatory processing/maturation of specific mRNAs, such as the T-box family members thrS and thrZ, but may also contribute to global mRNA degradation. PMID- 15831788 TI - DNA-binding specificity of rice mariner-like transposases and interactions with Stowaway MITEs. AB - Mariner-like elements (MLEs) are DNA transposons found throughout the plant and animal kingdoms. A previous computational survey of the rice (Oryza sativa) genome sequence revealed 34 full length MLEs (Osmars) belonging to 25 distinct families. This survey, which also identified sequence similarities between the Osmar elements and the Stowaway superfamily of MITEs, led to the formulation of a hypothesis whereby Stowaways are mobilized by OSMAR transposases. Here we investigate the DNA-binding activities and specificities of two OSMAR transposases, OSMAR5 and OSMAR10. Like other mariner-like transposases, the OSMARs bind specifically to the terminal inverted repeat (TIR) sequences of their encoding transposons. OSMAR5 binds DNA through a bipartite N-terminal domain containing two functionally separable helix-turn-helix motifs, resembling the paired domain of Tc1-like transposases and PAX transcription factors in metazoans. Furthermore, binding of the OSMARs is not limited to their own TIRs; OSMAR5 transposase can also interact in vitro with TIRs from closely related Osmar elements and with consensus TIRs of several Stowaway families mined from the rice genome sequence. These results provide the first biochemical evidence for a functional relationship between Osmar elements and Stowaway MITEs and lead us to suggest that there is extensive cross-talk among related but distinct transposon families co-existing in a single eukaryote genome. PMID- 15831789 TI - Eukaryotic RNases H1 act processively by interactions through the duplex RNA binding domain. AB - Ribonucleases H have mostly been implicated in eliminating short RNA primers used for initiation of lagging strand DNA synthesis. Escherichia coli RNase HI cleaves these RNA-DNA hybrids in a distributive manner. We report here that eukaryotic RNases H1 have evolved to be processive enzymes by attaching a duplex RNA-binding domain to the RNase H region. Highly conserved amino acids of the duplex RNA binding domain are required for processivity and nucleic acid binding, which leads to dimerization of the protein. The need for a processive enzyme underscores the importance in eukaryotic cells of processing long hybrids, most of which remain to be identified. However, long RNA-DNA hybrids formed during immunoglobulin class-switch recombination are potential targets for RNase H1 in the nucleus. In mitochondria, where RNase H1 is essential for DNA formation during embryogenesis, long hybrids may be involved in DNA replication. PMID- 15831790 TI - Masking repeats while clustering ESTs. AB - A problem in EST clustering is the presence of repeat sequences. To avoid false matches, repeats have to be masked. This can be a time-consuming process, and it depends on available repeat libraries. We present a fast and effective method that aims to eliminate the problems repeats cause in the process of clustering. Unlike traditional methods, repeats are inferred directly from the EST data, we do not rely on any external library of known repeats. This makes the method especially suitable for analysing the ESTs from organisms without good repeat libraries. We demonstrate that the result is very similar to performing standard repeat masking before clustering. PMID- 15831791 TI - Repair activity of base and nucleotide excision repair enzymes for guanine lesions induced by nitrosative stress. AB - Nitric oxide (NO) induces deamination of guanine, yielding xanthine and oxanine (Oxa). Furthermore, Oxa reacts with polyamines and DNA binding proteins to form cross-link adducts. Thus, it is of interest how these lesions are processed by DNA repair enzymes in view of the genotoxic mechanism of NO. In the present study, we have examined the repair capacity for Oxa and Oxa-spermine cross-link adducts (Oxa-Sp) of enzymes involved in base excision repair (BER) and nucleotide excision repair (NER) to delineate the repair mechanism of nitrosative damage to guanine. Oligonucleotide substrates containing Oxa and Oxa-Sp were incubated with purified BER and NER enzymes or cell-free extracts (CFEs), and the damage excising or DNA-incising activity was compared with that for control (physiological) substrates. The Oxa-excising activities of Escherichia coli and human DNA glycosylases and HeLa CFEs were 0.2-9% relative to control substrates, implying poor processing of Oxa by BER. In contrast, DNA containing Oxa-Sp was incised efficiently by UvrABC nuclease and SOS-induced E.coli CFEs, suggesting a role of NER in ameliorating genotoxic effects associated with nitrosative stress. Analyses of the activity of CFEs from NER-proficient and NER-deficient human cells on Oxa-Sp DNA confirmed further the involvement of NER in the repair of nitrosative DNA damage. PMID- 15831792 TI - Overexpression of Bcl-2 is associated with apoptotic resistance to the G quadruplex ligand 12459 but is not sufficient to confer resistance to long-term senescence. AB - The triazine derivative 12459 is a potent G-quadruplex interacting agent that inhibits telomerase activity. This agent induces time- and dose-dependent telomere shortening, senescence-like growth arrest and apoptosis in the human A549 tumour cell line. We show here that 12459 induces a delayed apoptosis that activates the mitochondrial pathway. A549 cell lines selected for resistance to 12459 and previously characterized for an altered hTERT expression also showed Bcl-2 overexpression. Transfection of Bcl-2 into A549 cells induced a resistance to the short-term apoptotic effect triggered by 12459, suggesting that Bcl-2 is an important determinant for the activity of 12459. In sharp contrast, the Bcl-2 overexpression was not sufficient to confer resistance to the senescence-like growth arrest induced by prolonged treatment with 12459. We also show that 12459 provokes a rapid degradation of the telomeric G-overhang in conditions that paralleled the apoptosis induction. In contrast, the G-overhang degradation was not observed when apoptosis was induced by camptothecin. Bcl-2 overexpression did not modify the G-overhang degradation, suggesting that this event is an early process uncoupled from the final apoptotic pathway. PMID- 15831793 TI - APE1-dependent repair of DNA single-strand breaks containing 3'-end 8-oxoguanine. AB - DNA single-strand breaks containing 3'-8-oxoguanine (3'-8-oxoG) ends can arise as a consequence of ionizing radiation and as a result of DNA polymerase infidelity by misincorporation of 8-oxodGMP. In this study we examined the mechanism of repair of 3'-8-oxoG within a single-strand break using purified base excision repair enzymes and human whole cell extracts. We find that 3'-8-oxoG inhibits ligation by DNA ligase IIIalpha or DNA ligase I, inhibits extension by DNA polymerase beta and that the lesion is resistant to excision by DNA glycosylases involved in the repair of oxidative lesions in human cells. However, we find that purified human AP-endonuclease 1 (APE1) is able to remove 3'-8-oxoG lesions. By fractionation of human whole cell extracts and immunoprecipitation of fractions containing 3'-8-oxoG excision activity, we further demonstrate that APE1 is the major activity involved in the repair of 3'-8-oxoG lesions in human cells and finally we reconstituted repair of the 3'-8-oxoG-containing oligonucleotide duplex with purified human enzymes including APE1, DNA polymerase beta and DNA ligase IIIalpha. PMID- 15831794 TI - Asymmetry in RNA pseudoknots: observation and theory. AB - RNA can fold into a topological structure called a pseudoknot, composed of non nested double-stranded stems connected by single-stranded loops. Our examination of the PseudoBase database of pseudoknotted RNA structures reveals asymmetries in the stem and loop lengths and provocative composition differences between the loops. By taking into account differences between major and minor grooves of the RNA double helix, we explain much of the asymmetry with a simple polymer physics model and statistical mechanical theory, with only one adjustable parameter. PMID- 15831795 TI - Pulmonary impedance and alveolar instability during injurious ventilation in rats. AB - The mechanical derangements in the acutely injured lung have long been ascribed, in large part, to altered mechanical function at the alveolar level. This has not been directly demonstrated, however, so we investigated the issue in a rat model of overinflation injury. After thoracotomy, rats were mechanically ventilated with either 1) high tidal volume (Vt) or 2) low Vt with periodic deep inflations (DIs). Forced oscillations were used to measure pulmonary impedance every minute, from which elastance (H) and hysteresivity (eta) were derived. Subpleural alveoli were imaged every 15 min using in vivo video microscopy. Cross-sectional areas of individual alveoli were measured at peak inspiration and end exhalation, and the percent change was used as an index of alveolar instability (%I-EDelta). Low Vt never led to an increase in %I-EDelta but did result in progressive atelectasis that coincided with an increase in H but not eta. DI reversed atelectasis due to low Vt, returning H to baseline. %I-EDelta, H, and eta all began to rise by 30 min of high Vt and were not reduced by DI. We conclude that simultaneous increases in both H and eta are reflective of lung injury in the form of alveolar instability, whereas an isolated and reversible increase in H during low Vt reflects merely derecruitment of alveoli. PMID- 15831796 TI - Ingestion of a high-glycemic index meal increases muscle glycogen storage at rest but augments its utilization during subsequent exercise. AB - The aim of this study was to compare the effect of preexercise breakfast containing high- and low-glycemic index (GI) carbohydrate (CHO) (2.5g CHO/kg body mass) on muscle glycogen metabolism. On two occasions, 14 days apart, seven trained men ran at 71% maximal oxygen uptake for 30 min on a treadmill. Three hours before exercise, in a randomized order, subjects consumed either isoenergetic high- (HGI) or low-GI (LGI) CHO breakfasts that provided (per 70 kg body mass) 3.43 MJ energy, 175 g CHO, 21 g protein, and 4 g fat. The incremental areas under the 3-h plasma glucose and serum insulin response curves after the HGI meal were 3.9- (P < 0.05) and 1.4-fold greater (P < 0.001), respectively, than those after the LGI meal. During the 3-h postprandial period, muscle glycogen concentration increased by 15% (P < 0.05) after the HGI meal but remained unchanged after the LGI meal. Muscle glycogen utilization during exercise was greater in the HGI (129.1 +/- 16.1 mmol/kg dry mass) compared with the LGI (87.9 +/- 15.1 mmol/kg dry mass; P < 0.01) trial. Although the LGI meal contributed less CHO to muscle glycogen synthesis in the 3-h postprandial period compared with the HGI meal, a sparing of muscle glycogen utilization during subsequent exercise was observed in the LGI trial, most likely as a result of better maintained fat oxidation. PMID- 15831797 TI - Effects of endurance exercise-training on single-fiber contractile properties of insulin-treated streptozotocin-induced diabetic rats. AB - The purpose of this study was to characterize the contractile properties of individual skinned muscle fibers from insulin-treated streptozotocin-induced diabetic rats after an endurance exercise training program. We hypothesized that single-fiber contractile function would decrease in the diabetic sedentary rats and that endurance exercise would preserve the function. In the study, 28 rats were assigned to either a nondiabetic sedentary, a nondiabetic exercise, a diabetic sedentary, or a diabetic exercise group. Rats in the diabetic groups received subcutaneous intermediate-lasting insulin daily. The exercise-trained rats ran on a treadmill at a moderate intensity for 60 min, five times per week. After 12 wk, the extensor digitorum longus and soleus muscles were dissected. Single-fiber diameter, Ca(2+)-activated peak force, specific tension, activation threshold, and pCa(50) as well as the myosin heavy chain isoform expression (MHC) were determined. We found that in MHC type II fibers from extensor digitorum longus muscle, diameters were significantly smaller from diabetic sedentary rats compared with nondiabetic sedentary rats (P < 0.001). Among the nondiabetic rats, fiber diameters were smaller with exercise (P = 0.038). The absolute force generating capacity of single fibers was lower in muscles from diabetic rats. There was greater specific tension (force normalized to cross-sectional area) by fibers from the rats that followed an endurance exercise program compared with sedentary. From the results, we conclude that alterations in the properties of contractile proteins are not implicated in the decrease in strength associated with diabetes and that endurance-exercise training does not prevent or increase muscle weakness in diabetic rats. PMID- 15831798 TI - Effects of a 6-mo endurance-training program on venous compliance and maximal lower body negative pressure in older men and women. AB - Aging and chronic exercise training influence leg venous compliance. Venous compliance affects responses to an orthostatic stress. The extent to which exercise training in a previously sedentary older population will affect venous compliance and tolerance to the simulated orthostatic stress of maximal lower body negative pressure (LBNP) is unknown. The purpose of this investigation is to determine the influence of a 6-mo endurance-training program on calf venous compliance and responses and tolerance to maximal LBNP in older men and women. Twenty participants (exercise group: n = 10, 5 men, 5 women; control group: n = 10, 6 men, 4 women; all >60 yr) underwent graded LBNP to presyncope or 4 min at 100 mmHg before and after a 6-mo endurance-training program. Utilizing venous occlusion plethysmography, calf venous compliance was determined in both groups using the first derivative of the pressure-volume relation during cuff pressure reduction before training, at 3 mo, and at the end of the training program. The exercise group improved their fitness with the 6-mo endurance-training program, whereas the control group did not change (14 +/- 3 vs. <1 +/- 2%; P < 0.05). LBNP tolerance did not differ between groups or across trials (P = 0.47). Venous compliance was not different between groups or trials, either initially or after 3 mo of endurance training, but tended to be greater in the exercise group after 6 mo of training (P = 0.08). These data suggest that a 6-mo endurance-training program may improve venous compliance without affecting tolerance to maximal LBNP in older participants. PMID- 15831799 TI - Chronic hypoxia exposure depresses aortic endothelium-dependent vasorelaxation in both sedentary and trained rats: involvement of L-arginine. AB - This study was designed to test the hypothesis that the previously demonstrated training-induced improvement of the endothelium vasodilator function would be blunted under conditions of chronic hypoxia exposure as a result of deleterious effects of hypoxia per se on the nitric oxide pathway. Sea-level-native rats were randomly assigned to N (living in normoxia), NT (living and training 5 days/wk for 5 wk in normoxia), CH (living in hypoxia, 2,800 m), and CHT (living and training 5 days/wk for 5 wk in hypoxia, 2,800 m) groups. Concentration-response curves to acetylcholine (ACh; 10(-9) to 10(-4) M) with or without L-arginine (10( 3) to 10(-5) M) and/or nitro-L-arginine methyl ester (10(-5) M) were assessed on aortic isolated rings. The main finding was that chronic hypoxia severely depressed maximal ACh-responses of aortic rings in both sedentary and trained groups. However, chronic hypoxia did not interfere with training-induced increases in maximal ACh responses, considering that maximal ACh vasorelaxation was improved in CHT rats to the same extent as in NT rats when both groups were directly compared with their sedentary counterparts. It should be pointed out that the vasodilator response to ACh was restored in CH and CHT rats to the level obtained in N and NT rats, respectively, by an in vitro L-arginine addition. A hypoxia-induced decrease in L-arginine bioavailability resulting from acclimatization at altitude may be involved in this limitation of the NO pathway in CH and CHT rats. These results are of importance for aerobic performance as the specific vascular adaptations to training at altitude could contribute to limit peripheral vasodilatation and subsequently blood flow during exercise. PMID- 15831800 TI - Computational simulation of human upper airway collapse using a pressure-/state dependent model of genioglossal muscle contraction under laminar flow conditions. AB - A three-element, pressure- and state (sleep and wake) -dependent contraction model of the genioglossal muscle was developed based on the microstructure of skeletal muscle and the cross-bridge theory. This model establishes a direct connection between the contractile forces generated in muscle fibers and the measured electromyogram signals during various upper airway conditions. This effectively avoids the difficulty of determining muscle shortening velocity during complex pharyngeal conditions when modeling the muscle's contractile behaviors. The activation of the genioglossal muscle under different conditions was then simulated. A sensitivity analysis was performed to determine the effects of varying each modeled parameter on the muscle's contractile behaviors. This muscle contraction model was then incorporated into our anatomically correct, two dimensional computational model of the pharyngeal airway to perform a finite element analysis of air flow, tissue deformation, and airway collapse. The model predicted muscle deformations are consistent with previous observations regarding upper airway behavior in normal subjects. PMID- 15831801 TI - Effects of selective hypoglossal nerve stimulation on canine upper airway mechanics. AB - Electrical stimulation of the hypoglossal (XII) nerve has been demonstrated as an effective approach to treating obstructive sleep apnea. The physiological effects of conventional modes of stimulation (i.e., genioglossus activation or whole XII nerve stimulation), however, have yielded inconsistent and only partial alleviations of hypopneic or apneic events. Although selective stimulation of the multifasciculated XII nerve offers many stimulus options, it is not clear how these will functionally affect the upper airway (UAW). To study these effects, animal experiments in eight beagles were performed to investigate changes in the UAW resistance and critical pressure during simulated expiration (n = 4) and inspiration (n = 4). During expiration, nonselective XII nerve stimulation yielded the greatest improvement in UAW resistance (-0.66 +/- 0.11 cm H2O x l(-1) x min(-1)), compared with that for selective activation of the geniohyoid (-0.29 +/- 0.09 cm H2O x l(-1) x min(-1)), genioglossus (-0.31 +/- 0.12 cm H2O x l(-1) x min(-1)), and hyoglossus/styloglossus (0.37 +/- 0.06 cm H2O x l(-1) x min(-1)) muscles. For simulated inspiration, on the other hand, only whole XII nerve stimulation (-0.9 +/- 0.4 cm H2O) and coactivation of the genioglossus + hyoglossus/styloglossus muscles (-1.18 +/- 0.6 cm H2O) produced significant (P < 0.05) improvements in UAW stability (i.e., lowered critical pressure), compared with baseline (-0.52 +/- 0.32 cm H2O). The results of this study suggest that a multicontact nerve electrode can be used to achieve both UAW dilation and patency, comparable to that obtained with nonselective stimulation, by selectively activating the various branches of the XII nerve. PMID- 15831802 TI - Caffeine increases exogenous carbohydrate oxidation during exercise. AB - Both carbohydrate (CHO) and caffeine have been used as ergogenic aids during exercise. It has been suggested that caffeine increases intestinal glucose absorption, but there are also suggestions that it may decrease muscle glucose uptake. The purpose of the study was to investigate the effect of caffeine on exogenous CHO oxidation. In a randomized crossover design, eight male cyclists (age 27 +/- 2 yr, body mass 71.2 +/- 2.3 kg, maximal oxygen uptake 65.7 +/- 2.2 ml x kg(-1) x min(-1)) exercised at 64 +/- 3% of maximal oxygen uptake for 120 min on three occasions. During exercise subjects ingested either a 5.8% glucose solution (Glu; 48 g/h), glucose with caffeine (Glu+Caf, 48 g/h + 5 mg x kg(-1) x h(-1)), or plain water (Wat). The glucose solution contained trace amounts of [U 13C]glucose so that exogenous CHO oxidation could be calculated. CHO and fat oxidation were measured by indirect calorimetry, and 13C appearance in the expired gases was measured by continuous-flow IRMS. Average exogenous CHO oxidation over the 90- to 120-min period was 26% higher (P < 0.05) in Glu+Caf (0.72 +/- 0.04 g/min) compared with Glu (0.57 +/- 0.04 g/min). Total CHO oxidation rates were higher (P < 0.05) in the CHO ingestion trials compared with Wat, but they were highest when Glu+Caf was ingested (1.21 +/- 0.37, 1.84 +/- 0.14, and 2.47 +/- 0.23 g/min for Wat, Glu, and Glu+Caf, respectively; P < 0.05). There was also a trend (P = 0.082) toward an increased endogenous CHO oxidation with Glu+Caf (1.81 +/- 0.22 g/min vs. 1.27 +/- 0.13 g/min for Glu and 1.12 +/- 0.37 g/min for Wat). In conclusion, compared with glucose alone, 5 mg x kg(-1) x h(-1) of caffeine coingested with glucose increases exogenous CHO oxidation, possibly as a result of an enhanced intestinal absorption. PMID- 15831803 TI - Early effects of mechanical ventilation on isotonic contractile properties and MAF-box gene expression in the diaphragm. AB - This study aimed to determine the time-dependent effects of diaphragmatic inactivity on its maximum shortening velocity (V(max)) and the muscle atrophy F box (MAF-box, atrogin-1) gene expression during controlled mechanical ventilation (CMV). Twenty-four New Zealand White rabbits were grouped into 1 day, 2 days, and 3 days of CMV and controls in equal numbers. The in vitro isotonic contractile properties of the diaphragm were determined. In addition, myosin heavy chain protein and mRNA, myosin light chain, MAF-box mRNA, and volume density of abnormal myofibrils were measured. Tetanic force decreased, and V(max) increased from control of 6.4 to 6.6, 7.7, and 8.1 muscle lengths per second after 1, 2, and 3 days of CMV, respectively (P < 0.02). The increased V(max) compensated for the decreased tetanic force; consequently, compared with the controls, maximum power output was unchanged after 3 days of CMV. V(max) correlated with the volume density of abnormal myofibrils [y = 0.1x + 5.7 (r = 0.87, P < 0.01)]. In the diaphragm, MAF-box was overexpressed (355% of control) after 1 day of CMV, before the evidence of structural myofibril disarray. In conclusion, CMV produced a time dependent increase in V(max) that was associated with the degree of myofibrillar disarray and independent of changes in myosin isoform expression. Furthermore, CMV produced an increase in MAF-box mRNA levels that may be partially or completely responsible for the degree of myofibrillar disarray resulting from CMV. PMID- 15831804 TI - Metabolic equivalent: one size does not fit all. AB - The metabolic equivalent (MET) is a widely used physiological concept that represents a simple procedure for expressing energy cost of physical activities as multiples of resting metabolic rate (RMR). The value equating 1 MET (3.5 ml O2 x kg(-1) x min(-1) or 1 kcal x kg(-1) x h(-1)) was first derived from the resting O2 consumption (VO2) of one person, a 70-kg, 40-yr-old man. Given the extensive use of MET levels to quantify physical activity level or work output, we investigated the adequacy of this scientific convention. Subjects consisted of 642 women and 127 men, 18-74 yr of age, 35-186 kg in weight, who were weight stable and healthy, albeit obese in some cases. RMR was measured by indirect calorimetry using a ventilated hood system, and the energy cost of walking on a treadmill at 5.6 km/h was measured in a subsample of 49 men and 49 women (26-45 kg/m2; 29-47 yr). Average VO2 and energy cost corresponding with rest (2.6 +/- 0.4 ml O2 x kg(-1) x min(-1) and 0.84 +/- 0.16 kcal x kg(-1) x h(-1), respectively) were significantly lower than the commonly accepted 1-MET values of 3.5 ml O2 x kg(-1) x min(-1) and 1 kcal x kg(-1) x h(-1), respectively. Body composition (fat mass and fat-free mass) accounted for 62% of the variance in resting VO2 compared with age, which accounted for only 14%. For a large heterogeneous sample, the 1-MET value of 3.5 ml O2 x kg(-1) x min(-1) overestimates the actual resting VO2 value on average by 35%, and the 1-MET of 1 kcal/h overestimates resting energy expenditure by 20%. Using measured or predicted RMR (ml O2 x kg(-1) x min(-1) or kcal x kg(-1) x h(-1)) as a correction factor can appropriately adjust for individual differences when estimating the energy cost of moderate intensity walking (5.6 km/h). PMID- 15831805 TI - Phagocytosis of apoptotic cells by macrophages is impaired in atherosclerosis. AB - OBJECTIVE: Apoptotic cell death has been demonstrated in advanced human atherosclerotic plaques. Apoptotic cells (ACs) should be rapidly removed by macrophages, otherwise secondary necrosis occurs, which in turn elicits inflammatory responses and plaque progression. Therefore, we investigated the efficiency of phagocytosis of ACs by macrophages in atherosclerosis. METHODS AND RESULTS: Human endarterectomy specimens and human tonsils were costained for CD68 (macrophages) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) (apoptosis). Free and phagocytized ACs were counted in both tissues. The ratio of free versus phagocytized AC was 19-times higher in human atherosclerotic plaques as compared with human tonsils, indicating a severe defect in clearance of AC. Impaired phagocytosis of AC was also detected in plaques from cholesterol-fed rabbits and did not further change with plaque progression. In vitro experiments with J774 or peritoneal mouse macrophages showed that several factors caused impaired phagocytosis of AC including cytoplasmic overload of macrophages with indigestible material (beads), free radical attack, and competitive inhibition among oxidized red blood cells, oxidized low-density lipoprotein and ACs for the same receptor(s) on the macrophage. CONCLUSIONS: Our data demonstrate that phagocytosis of ACs is impaired in atherosclerotic plaques, which is at least partly attributed to oxidative stress and cytoplasmic saturation with indigestible material. PMID- 15831806 TI - ACAT2 is a target for treatment of coronary heart disease associated with hypercholesterolemia. AB - The inhibition of intracellular cholesterol esterification as a means to prevent atherosclerosis has been considered to have potential for many years. Two different ACAT enzymes were discovered about 7 years ago, and it has become clear that the two enzymes provide separate physiologic functions. Much has been learned from mice with gene deletions for either ACAT1 or ACAT2. Deletion of ACAT2 has consistently been atheroprotective whereas deletion of ACAT1 has been varyingly problematic. ACAT1 functions in converting cellular cholesterol into cholesteryl ester in response to cholesterol abundance inside the cells. In atherosclerotic lesions, where macrophages ingest excess cholesterol, the ability to esterify the newly-acquired cholesterol seems important for cell survival. Inhibition of ACAT1 may bring undesired consequences with destabilization of cellular membrane function upon cholesterol accumulation leading to macrophage cell death. In contrast, ACAT2 is expressed only in hepatocytes and enterocytes, where ACAT1 is silent, and appears to provide cholesteryl esters for transport in lipoproteins. These two cell types have an abundance of additional mechanisms for disposing of cholesterol so that depletion of ACAT2 does not signal apoptosis. At the present time, the bulk of the available data suggest that the strategy seeming to bear the most potential for treatment of coronary heart disease associated with hypercholesterolemia would be to specifically inhibit ACAT2. PMID- 15831807 TI - Increased cholesterol deposition, expression of scavenger receptors, and response to chemotactic factors in Abca1-deficient macrophages. AB - OBJECTIVE: Studies in bone marrow transplanted from ATP-binding cassette transporter A1 (ABCA1)-deficient mice into normal mice provides direct evidence that the absence of leukocyte ABCA1 exerts a marked proatherogenic effect independent of changes in plasma lipids, suggesting that ABCA1 plays a key role in the regulation of cholesterol homeostasis and function of macrophages. Therefore, we examined whether the absence of ABCA1 affects the morphology, properties, and functional activities of macrophages that could be related to the development of atherosclerosis. METHODS AND RESULTS: We conducted a series of experiments in macrophages isolated from Abca1-deficient and wild-type mice and compared several of their properties that are thought to be related to the development of atherosclerosis. Macrophages isolated from Abca1-deficient mice have an increase in cholesterol content, expression of scavenger receptors, and secretion of chemokines, growth factors, and cytokines, resulting in an increased ability to respond to a variety of chemotactic factors. CONCLUSIONS: Our studies indicate that the absence of ABCA1 leads to significant changes in the morphology, properties, and functional activities of macrophages. These changes, together with the proinflammatory condition present in ABCA1-deficient mice and increased reactivity of macrophages to chemotactic factors, play a key role in the development and progression of atherosclerosis. PMID- 15831808 TI - New links between inflammation and thrombosis. AB - This article is a summary of the Sol Sherry Lecture of the Council on Arteriosclerosis, Thrombosis, and Vascular Biology, which was presented at the Scientific Sessions of the American Heart Association in November 2004. It highlights work from our laboratory, focusing mainly on new aspects of P-selectin and CD40L (CD154) biology and on the interplay of platelets and leukocytes in thrombosis and inflammation. PMID- 15831809 TI - Humoral immune response against defined oxidized low-density lipoprotein antigens reflects structure and disease activity of carotid plaques. AB - BACKGROUND: Immune responses against oxidized low-density lipoprotein (LDL) play an important role in atherosclerosis. The aim of this study was to investigate if humoral immune response against specific oxidized LDL antigens, such as aldehyde modified peptide sequences of apolipoprotein B-100, reflects disease activity and structure of atherosclerotic plaques. METHODS AND RESULTS: Plaques were obtained from 114 symptomatic subjects referred to carotid endarterectomy and characterized immunohistochemically and histologically. Plasma levels of IgG and IgM against aldehyde-modified apolipoprotein B-100 amino acid sequences 661 to 680, 3136 to 3155 (peptide 210), and 3661 to 3680 (peptide 240) were determined by enzyme-linked immunosorbent assay. High levels of IgG against peptide 210 were associated with increased plaque content of lipids (r=0.24, P<0.05) and hemorrhage (r=0.27, P=0.005), with decreased content of fibrous tissue (r=-0.25, P=0.01), but also with lower total plaque volume (r=-0.21, P<0.05). In contrast, high levels of IgM against peptide 240 were associated with plaques with more fibrous tissue (r=0.35, P<0.001), less lipids (r=-0.34, P<0.001), and less macrophages (r=-0.24, P<0.05). IgM against peptide 210 were found to be associated with plaque fibrous tissue (r=0.20, P<0.05), less lipids (r=-0.21, P<0.05), and less macrophages (r=-0.27, P=0.01). CONCLUSIONS: These findings support the notion that immune responses against oxidized LDL epitopes are involved in atherosclerosis and that the level of circulating antibodies against these structures may reflect disease activity in the arterial wall. PMID- 15831810 TI - Systemic endothelial dysfunction as an early predictor of adverse outcome in heart failure. AB - OBJECTIVE: Endothelial dysfunction is an early event in the natural progression of heart failure. Increased oxidative stress has been linked to impaired endothelial function and both may play a prognostic role. METHODS AND RESULTS: Endothelium-dependent and endothelium-independent vasodilatation were determined in 289 patients with mild left ventricular dysfunction by measuring forearm blood flow responses to acetylcholine and sodium nitroprusside using venous occlusion plethysmography. Vascular effects of the coadministration of the antioxidant vitamin C at pharmacological doses (24 mg/min) were assessed. Occurrence of death, heart transplantation, and readmission with worsening heart failure were recorded as clinical outcome parameters during a follow-up period of 4.8 years. Patients experiencing adverse events (n=79) had lower vasodilator responses to acetylcholine (P<0.001) and to sodium nitroprusside (P=0.03) compared with patients without events. However, beneficial effects of vitamin C did not differ between both groups. Cox proportional hazards model demonstrated that age (P=0.001), renal function (P=0.001), and blunted acetylcholine-induced vasodilatation (P=0.007) remained independent predictors of adverse outcome. CONCLUSIONS: Impaired peripheral endothelial function independently predicts long term adverse outcome in patients with early-stage heart failure. The findings suggest that assessment of peripheral endothelial function may represent an additional mean for risk stratification and therapy management in these patients. PMID- 15831811 TI - Vascular endothelial growth factor-expressing mesenchymal stem cell transplantation for the treatment of acute myocardial infarction. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF) plays an important role in inducing angiogenesis. Mesenchymal stem cells (MSCs) may have potential for differentiation to several types of cells, including myocytes. We hypothesized that transplantation of VEGF-expressing MSCs could effectively treat acute myocardial infarction (MI) by providing enhanced cardioprotection, followed by angiogenic effects in salvaging ischemic myocardium. METHODS AND RESULTS: The human VEGF165 gene was transfected to cultured MSCs of Lewis rats using an adenoviral vector. Six million VEGF-transfected and LacZ-transfected MSCs (VEGF group), LacZ-transfected MSCs (control group), or serum-free medium only (medium group) were injected into syngeneic rat hearts 1 hour after left coronary artery occlusion. At 1 week after MI, MSCs were detected by X-gal staining in infarcted region. High expression of VEGF was immunostained in the VEGF group. At 28 days after MI, infarct size, left ventricular dimensions, ejection fraction, E wave/A wave ratio and capillary density of the infarcted region were most improved in the VEGF group, compared with the medium group. Immunohistochemically, alpha smooth muscle actin-positive cells were most increased in the VEGF group. CONCLUSIONS: This combined strategy of cell transplantation with gene therapy could be a useful therapy for the treatment of acute MI. PMID- 15831812 TI - Apolipoprotein A-I mimetic peptides. AB - Despite identical amino acid composition, differences in class A amphipathic helical peptides caused by differences in the order of amino acids on the hydrophobic face results in substantial differences in antiinflammatory properties. One of these peptides is an apolipoprotein A-I (apoA-I) mimetic, D 4F. When given orally to mice and monkeys, D-4F caused the formation of pre-beta high-density lipoprotein (HDL), improved HDL-mediated cholesterol efflux, reduced lipoprotein lipid hydroperoxides, increased paraoxonase activity, and converted HDL from pro-inflammatory to antiinflammatory. In apolipoprotein E (apoE)-null mice, D-4F increased reverse cholesterol transport from macrophages. Oral D-4F reduced atherosclerosis in apoE-null and low-density lipoprotein (LDL) receptor null mice. In vitro when added to human plasma at nanomolar concentrations, D-4F caused the formation of pre-beta HDL, reduced lipoprotein lipid hydroperoxides, increased paraoxonase activity, and converted HDL from pro-inflammatory to antiinflammatory. Physical-chemical properties and the ability of various class A amphipathic helical peptides to activate lecithin cholesterol acyltransferase (LCAT) in vitro did not predict biologic activity in vivo. In contrast, the use of cultured human artery wall cells in evaluating these peptides was more predictive of their efficacy in vivo. We conclude that the antiinflammatory properties of different class A amphipathic helical peptides depends on subtle differences in the configuration of the hydrophobic face of the peptides, which determines the ability of the peptides to sequester inflammatory lipids. These differences appear to be too subtle to predict efficacy based on physical chemical properties alone. However, understanding these physical-chemical properties provides an explanation for the mechanism of action of the active peptides. PMID- 15831813 TI - Regulation of vascular smooth muscle cell proliferation: role of NF-kappaB revisited. AB - The transcription factor NF-kappaB regulates cell cycle progression and proliferation in a number of cell types. An important unresolved issue is the potential role of NF-kappaB in the proliferation of vascular smooth muscle cells (VSMCs) as a basis for the development of vascular disease. To investigate the contribution of NF-kappaB to mitogen-induced proliferation of VSMCs, a knock-in mouse model expressing the NF-kappaB superrepressor IkappaBalphaDeltaN (c(IkappaBalphaDeltaN)) was used. Comparing wild-type and IkappaBalphaDeltaN expressing VSMCs, we found that proliferation rates did not differ after mitogenic stimulation by platelet-derived growth-factor-BB (PDGF-BB) or serum. In line with this, NF-kappaB activation was not observed in VSMCs derived from transgenic mice expressing an NF-kappaB-dependent lacZ reporter (c((Igk)3conalacZ)). We further show, that classical mitogenic signaling pathways (namely mitogen-activated protein kinase [MAPK] and the phosphatidyl-inositol-3 OH-kinase [PI3K] pathways) control VSMC proliferation, but independently of NF kappaB activation. In contrast to VSMCs, mouse embryonic fibroblasts (MEFs) derived from IkappaBalphaDeltaN-expressing mice showed significantly impaired proliferation rates after mitogenic stimulation. This was reflected by strongly impaired cyclin D1 expression in serum-stimulated MEFs derived from (c(IkappaBalphaDeltaN)) mice. These results implicate that essential pathogenetic functions of NF-kappaB in the development of atherosclerosis involve apoptotic and inflammatory signaling of VSMCs rather than proliferation. They further provide genetic evidence for a cell-type restricted requirement of NF-kappaB in the control of cellular proliferation. PMID- 15831814 TI - Angiotensin IV activates the nuclear transcription factor-kappaB and related proinflammatory genes in vascular smooth muscle cells. AB - Inflammation is a key event in the development of atherosclerosis. Nuclear factor kappaB (NF-kappaB) is important in the inflammatory response regulation. The effector peptide of the renin angiotensin system Angiotensin II (Ang II) activates NF-kappaB and upregulates some related proinflammatory genes. Our aim was to investigate whether other angiotensin-related peptides, as the N-terminal degradation peptide Ang IV, could regulate proinflammatory factors (activation of NF-kappaB and related genes) in cultured vascular smooth muscle cells (VSMCs). In these cells, Ang IV increased NF-kappaB DNA binding activity, caused nuclear translocation of p50/p65 subunits, cytosolic IkappaB degradation and induced NF kappaB-dependent gene transcription. Ang II activates NF-kappaB via AT1 and AT2 receptors, but AT1 or AT2 antagonists did not inhibit NF-kappaB activation caused by Ang IV. In VSMC from AT1a receptor knockout mice, Ang IV also activated NF kappaB pathway. In those cells, the AT4 antagonist divalinal diminished dose dependently Ang IV-induced NF-kappaB activation and prevented IkappaB degradation, but had no effect on the Ang II response, indicating that Ang IV activates the NF-kappaB pathway via AT4 receptors. Ang IV also increased the expression of proinflammatory factors under NF-kappaB control, such as MCP-1, IL 6, TNF-alpha, ICAM-1, and PAI-1, which were blocked by the AT4 antagonist. Our results reveal that Ang IV, via AT4 receptors, activates NF-kappaB pathway and increases proinflammatory genes. These data indicate that Ang IV possesses proinflammatory properties, suggesting that this Ang degradation peptide could participate in the pathogenesis of cardiovascular diseases. PMID- 15831815 TI - Role of histamine produced by bone marrow-derived vascular cells in pathogenesis of atherosclerosis. AB - To clarify the role of histamine-producing cells and its origin in atherosclerosis, we investigated histidine decarboxylase (HDC; histamine producing enzyme) expression in murine arteries with vascular injuries after the animal had received transplanted bone marrow (BM) from green fluorescent protein (GFP)-transgenic mice. The neointima in the ligated carotid arteries contained BM derived HDC+ cells that expressed macrophage (Mac-3) or smooth muscle cell antigen (alpha-SMA). In contrast, the HDC+ BM-derived cells, which were positive for Mac-3, were mainly located in the adventitia in the cuff replacement model. In apolipoprotein E-knockout mice on a high cholesterol diet, BM-derived cells expressing Mac-3 in the atheromatous plaques were also positive for HDC. In comparison with wild-type mice, HDC-/- mice showed reduced neointimal thickening and a decreased intima-to-media ratio after ligation and cuff replacement. These results indicate that histamine produced from BM-derived progenitor cells, which could transdifferentiate into SMC- or macrophage-like cells, are important for the formation of neointima and atheromatous plaques. PMID- 15831816 TI - Modulation of the cardiac sodium channel NaV1.5 by Fyn, a Src family tyrosine kinase. AB - Dynamic modulation of ion channels can produce dramatic alterations of electrical excitability in cardiac myocytes. This study addresses the effects of the Src family tyrosine kinase Fyn on Na(V)1.5 cardiac sodium channels. Sodium currents were acquired by whole cell recording on HEK-293 cells transiently expressing Na(V)1.5. Acute treatment of cells with insulin caused a depolarizing shift in steady-state inactivation, an effect eliminated by the Src-specific tyrosine kinase inhibitor PP2. Sodium channels were coexpressed with either constitutively active (Fyn(CA)) or catalytically inactive (Fyn(KD)) variants of Fyn. Fyn(CA) caused a 10-mV depolarizing shift of steady-state inactivation compared with Fyn(KD) without altering the activation conductance-voltage relationship. Comparable effects of these Fyn variants were obtained with whole-cell and perforated-patch recording. Tyrosine phosphorylation of immunoprecipitated Na(V)1.5 was increased in cells expressing Fyn(CA) compared with Fyn(KD). We show that Fyn is present in rat cardiac myocytes, and that Na(V)1.5 channels from these myocytes are tyrosine-phosphorylated. In HEK-293 cells the effect of Fyn(CA) on Na(V)1.5 inactivation is abolished by the single point mutation Y1495F, a residue located within the cytoplasmic linker between the third and fourth homologous domains of the sodium channel. We provide evidence that this linker is a substrate for Fyn in vitro, and that Y1495 is a preferred phosphorylation site. These results suggest that cardiac sodium channels are physiologically relevant targets of Src family tyrosine kinases. PMID- 15831818 TI - Novel mitogenic effect of adenosine on coronary artery smooth muscle cells: role for the A1 adenosine receptor. AB - Adenosine is a vascular endothelial cell mitogen, but anti-mitogenic for aortic smooth muscle cells and fibroblasts when acting via the A2B adenosine receptor. However, we show that adenosine increases porcine coronary artery smooth muscle cell (CASMC) number, cellular DNA content, protein synthesis, and PCNA staining. RT-PCR analysis indicates that porcine CASMC express A1, A2A, A3, and barely detectable levels of A2B receptor mRNAs. The mitogenic effect of adenosine is mimicked by NECA, CCPA, and R-PIA, but not by CGS21680and 2-Cl-IB-MECA, and is inhibited by DPCPX, indicating a prominent role for the A1 receptor. This interpretation is supported by the finding that adenosine- and CCPA-induced DNA synthesis is significantly inhibited by pertussis toxin, but substantially potentiated by PD81723, an allosteric enhancer of the A1 receptor. When a cDNA encoding the porcine A1 receptor was cloned and expressed in COS-1 cells, A1 receptor pharmacology is confirmed. Anti-sense oligonucleotides to the cloned sequence dramatically suppress the mitogenic effect of adenosine and CCPA. Conversely, over-expression of the cloned A1 receptor in CASMC increases adenosine- and CCPA-induced DNA synthesis. Furthermore, stimulation with adenosine or CCPA of intact coronary arteries in an organ culture model of vascular disease increases cellular DNA synthesis, which was abolished by DPCPX. We conclude that adenosine acts as a novel mitogen in porcine CASMC that express the A1 adenosine receptor, possibly contributing to the development of coronary artery disease. PMID- 15831819 TI - Short QT syndrome or Andersen syndrome: Yin and Yang of Kir2.1 channel dysfunction. PMID- 15831817 TI - Adenovirus vector E4 gene regulates connexin 40 and 43 expression in endothelial cells via PKA and PI3K signal pathways. AB - Connexins (Cxs) provide a means for intercellular communication and play important roles in the pathophysiology of vascular cardiac diseases. Infection of endothelial cells (ECs) with first-generation E1/E3-deleted E4+ adenovirus (AdE4+) selectively modulates the survival and angiogenic potential of ECs by as of yet unrecognized mechanisms. We show here that AdE4+ vectors potentiate Cx expression in ECs in vitro and in mouse heart tissue. Infection of ECs with AdE4+, but not AdE4-, resulted in a time- and dose-dependent induction of junctional Cx40 expression and suppression of Cx43 protein and mRNA expression. Treatment of ECs with PKA inhibitor H89 or PI3K inhibitor LY294002 prevented the AdE4+-mediated regulation of Cx40 and Cx43 that was associated with diminished AdE4+-mediated survival of ECs. Moreover, both PKA activity and cAMP-response element (CRE)-binding activity were enhanced by treatment of ECs with AdE4+. However, there is no causal evidence of a cross-talk between the 2 modulatory pathways, PKA and PI3K. Remarkably, Cx40 immunostaining was markedly increased and Cx43 was decreased in the heart tissue of mice treated with intra-tracheal AdE4+. Taken together, these results suggest that AdE4+ may play an important role in the regulation of Cx expression in ECs, and that these effects are mediated by both the PKA/CREB and PI3K signaling pathways. PMID- 15831820 TI - FGF induces hypertrophy and angiogenesis in hibernating myocardium. PMID- 15831821 TI - Targeting protein phosphatase 1 in heart failure. PMID- 15831822 TI - Cardiac hypertrophy served with protein kinase Cepsilon: delta isoform substitution available at additional cost. PMID- 15831823 TI - Regulation of vascular calcification: roles of phosphate and osteopontin. AB - Vascular calcification is prevalent in aging as well as a number of pathological conditions, and it is now recognized as a strong predictor of cardiovascular events in the general population as well as diabetic and end-stage renal disease patients. Vascular calcification is a highly regulated process involving inductive and inhibitory mechanisms. This article focuses on two molecules, phosphate and osteopontin, that have been implicated in the induction or inhibition of vascular calcification, respectively. Elevated phosphate is of interest because hyperphosphatemia is recognized as a major nonconventional risk factor for cardiovascular disease mortality in end-stage renal disease patients. Studies to date suggest that elevated phosphate stimulates smooth muscle cell phenotypic transition and mineralization via the activity of a sodium-dependent phosphate cotransporter. Osteopontin, however, appears to block vascular calcification most likely by preventing calcium phosphate crystal growth and inducing cellular mineral resorption. PMID- 15831824 TI - Clostridium difficile toxins: mechanism of action and role in disease. AB - As the leading cause of hospital-acquired diarrhea, Clostridium difficile colonizes the large bowel of patients undergoing antibiotic therapy and produces two toxins, which cause notable disease pathologies. These two toxins, TcdA and TcdB, are encoded on a pathogenicity locus along with negative and positive regulators of their expression. Following expression and release from the bacterium, TcdA and TcdB translocate to the cytosol of target cells and inactivate small GTP-binding proteins, which include Rho, Rac, and Cdc42. Inactivation of these substrates occurs through monoglucosylation of a single reactive threonine, which lies within the effector-binding loop and coordinates a divalent cation critical to binding GTP. By glucosylating small GTPases, TcdA and TcdB cause actin condensation and cell rounding, which is followed by death of the cell. TcdA elicits effects primarily within the intestinal epithelium, while TcdB has a broader cell tropism. Important advances in the study of these toxins have been made in the past 15 years, and these are detailed in this review. The domains, subdomains, and residues of these toxins important for receptor binding and enzymatic activity have been elegantly studied and are highlighted herein. Furthermore, there have been major advances in defining the role of these toxins in modulating the inflammatory events involving the disruption of cell junctions, neuronal activation, cytokine production, and infiltration by polymorphonuclear cells. Collectively, the present review provides a comprehensive update on TcdA and TcdB's mechanism of action as well as the role of these toxins in disease. PMID- 15831826 TI - Subversion mechanisms by which Leishmania parasites can escape the host immune response: a signaling point of view. AB - The obligate intracellular parasite Leishmania must survive the antimicrobial activities of its host cell, the macrophage, and prevent activation of an effective immune response. In order to do this, it has developed numerous highly successful strategies for manipulating activities, including antigen presentation, nitric oxide and oxygen radical generation, and cytokine production. This is generally the result of interactions between Leishmania cell surface molecules, particularly gp63 and LPG, and less well identified macrophage surface receptors, causing the distortion of specific intracellular signaling cascades. We describe some of the signaling pathways and intermediates that are repressed in infected cells, including JAK/STAT, Ca(2+)-dependent protein kinase C (PKC) isoforms, and mitogen-activated protein kinases (especially ERK1/2), and proteasome-mediated transcription factor degradation. We also discuss protein tyrosine phosphatases (particularly SHP-1), intracellular Ca2+, Ca(2+) independent PKC, ceramide, and the suppressors of cytokine signaling family of repressors, which are all reported to be activated following infection, and the role of parasite-secreted cysteine proteases. PMID- 15831825 TI - Pathogenesis of Afa/Dr diffusely adhering Escherichia coli. AB - Over the last few years, dramatic increases in our knowledge about diffusely adhering Escherichia coli (DAEC) pathogenesis have taken place. The typical class of DAEC includes E. coli strains harboring AfaE-I, AfaE-II, AfaE-III, AfaE-V, Dr, Dr-II, F1845, and NFA-I adhesins (Afa/Dr DAEC); these strains (i) have an identical genetic organization and (ii) allow binding to human decay-accelerating factor (DAF) (Afa/Dr(DAF) subclass) or carcinoembryonic antigen (CEA) (Afa/Dr(CEA) subclass). The atypical class of DAEC includes two subclasses of strains; the atypical subclass 1 includes E. coli strains that express AfaE-VII, AfaE-VIII, AAF-I, AAF-II, and AAF-III adhesins, which (i) have an identical genetic organization and (ii) do not bind to human DAF, and the atypical subclass 2 includes E. coli strains that harbor Afa/Dr adhesins or others adhesins promoting diffuse adhesion, together with pathogenicity islands such as the LEE pathogenicity island (DA-EPEC). In this review, the focus is on Afa/Dr DAEC strains that have been found to be associated with urinary tract infections and with enteric infection. The review aims to provide a broad overview and update of the virulence aspects of these intriguing pathogens. Epidemiological studies, diagnostic techniques, characteristic molecular features of Afa/Dr operons, and the respective role of Afa/Dr adhesins and invasins in pathogenesis are described. Following the recognition of membrane-bound receptors, including type IV collagen, DAF, CEACAM1, CEA, and CEACAM6, by Afa/Dr adhesins, activation of signal transduction pathways leads to structural and functional injuries at brush border and junctional domains and to proinflammatory responses in polarized intestinal cells. In addition, uropathogenic Afa/Dr DAEC strains, following recognition of beta(1) integrin as a receptor, enter epithelial cells by a zipper like, raft- and microtubule-dependent mechanism. Finally, the presence of other, unknown virulence factors and the way that an Afa/Dr DAEC strain emerges from the human intestinal microbiota as a "silent pathogen" are discussed. PMID- 15831827 TI - Metallo-beta-lactamases: the quiet before the storm? AB - The ascendancy of metallo-beta-lactamases within the clinical sector, while not ubiquitous, has nonetheless been dramatic; some reports indicate that nearly 30% of imipenem-resistant Pseudomonas aeruginosa strains possess a metallo-beta lactamase. Acquisition of a metallo-beta-lactamase gene will invariably mediate broad-spectrum beta-lactam resistance in P. aeruginosa, but the level of in vitro resistance in Acinetobacter spp. and Enterobacteriaceae is less dependable. Their clinical significance is further embellished by their ability to hydrolyze all beta-lactams and by the fact that there is currently no clinical inhibitor, nor is there likely to be for the foreseeable future. The genes encoding metallo-beta lactamases are often procured by class 1 (sometimes class 3) integrons, which, in turn, are embedded in transposons, resulting in a highly transmissible genetic apparatus. Moreover, other gene cassettes within the integrons often confer resistance to aminoglycosides, precluding their use as an alternative treatment. Thus far, the metallo-beta-lactamases encoded on transferable genes include IMP, VIM, SPM, and GIM and have been reported from 28 countries. Their rapid dissemination is worrisome and necessitates the implementation of not just surveillance studies but also metallo-beta-lactamase inhibitor studies securing the longevity of important anti-infectives. PMID- 15831831 TI - Gavestinel does not improve outcome after acute intracerebral hemorrhage: an analysis from the GAIN International and GAIN Americas studies. AB - BACKGROUND AND PURPOSE: Glycine Antagonist in Neuroprotection (GAIN) International and GAIN Americas trials were prospectively designed, randomized, placebo-controlled trials of gavestinel, a glycine-site antagonist and putative neuroprotectant drug administered within 6 hours of suspected ischemic or hemorrhagic stroke. Both trials reported that gavestinel was ineffective in ischemic stroke. This analysis reports the results in those with primary intracerebral hemorrhage. METHODS: The primary hypothesis was that gavestinel treatment did not alter outcome, measured at 3 months by the Barthel Index (BI), from acute intracerebral hemorrhage, based on pooled results from both trials. The BI scores were divided into 3 groups: 95 to 100 (independent), 60 to 90 (assisted independence), and 0 to 55 (dependent) or dead. RESULTS: In total, 3450 patients were randomized in GAIN International (N=1804) and GAIN Americas (N=1646). Of these, 571 were ultimately identified to have spontaneous intracerebral hematoma on baseline head computerized tomography scan. The difference in distribution of trichotomized BI scores at 3 months between gavestinel and placebo was not statistically significant (P=0.09). Serious adverse events were reported at similar rates in the 2 treatment groups. CONCLUSIONS: These observations from the combined GAIN International and GAIN Americas trials suggest that gavestinel is not of substantial benefit or harm to patients with primary intracerebral hemorrhage. These findings are similar to results previously reported in patients with ischemic stroke. PMID- 15831830 TI - Diagnosis and management of pediatric urinary tract infections. AB - Urinary tract infection (UTI) is among the most commonly diagnosed bacterial infections of childhood. Although frequently encountered and well researched, diagnosis and management of UTI continue to be a controversial issue with many challenges for the clinician. Prevalence studies have shown that UTI may often be missed on history and physical examination, and the decision to screen for UTI must balance the risk for missed infections with the cost and inconvenience of testing. Interpretation of rapid diagnostic tests and culture is complicated by issues of contamination, false test results, and asymptomatic colonization of the urinary tract with nonpathogenic bacteria. The appropriate treatment of UTI has been controversial and has become more complex with the emergence of resistance to commonly used antibiotics. Finally, the anatomic evaluation and long-term management of a child after a UTI have been based on limited evidence, and newer studies question some of the tenets of prior recommendations. The goal of this review is to provide an up-to-date summary of the literature with particular attention to practical questions about diagnosis and management for the clinician. PMID- 15831832 TI - Cerebrovascular health disparities. PMID- 15831835 TI - Use of functional MRI to guide decisions in a clinical stroke trial. AB - BACKGROUND AND PURPOSE: An investigational trial examined safety and efficacy of targeted subthreshold cortical stimulation in patients with chronic stroke. The anatomical location for the target, hand motor area, varies across subjects, and so was localized with functional MRI (fMRI). This report describes the experience of incorporating standardized fMRI into a multisite stroke trial. METHODS: At 3 enrollment centers, patients moved (0.25 Hz) the affected hand during fMRI. Hand motor function was localized at a fourth center guiding intervention for those randomized to stimulation. RESULTS: The fMRI results were available within 24 hours. Across 12 patients, activation site variability was substantial (12, 23, and 11 mm in x, y, and z directions), exceeding stimulating electrode dimensions. CONCLUSIONS: Use of fMRI to guide decision-making in a clinical stroke trial is feasible. PMID- 15831836 TI - Patterns of cerebral infarction in aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The aim of this study was to analyze the distribution patterns of delayed cerebral ischemia after subarachnoid hemorrhage (SAH) and the factors that determine their occurrence. METHODS: We analyzed clinical and radiological data of 143 consecutive patients with aneurysmal SAH. Computed tomography scan revision was blinded to clinical information. Superficial infarctions were defined as territorial lesions with cortical involvement. Perioperative infarctions were excluded. RESULTS: Fifty-six patients (39%) had cerebral infarctions. They were unilateral in 34 patients (61%) and involved a single territory in 29 (52%). Location was cortical in 34 patients (61%), deep in 10 (18%), and combined cortical and deep in 12 (21%). Single infarctions were cortical in 23 of 28 cases (79%). Deep territory ischemia was more common with multiple lesions (16/28, 57% versus 6/29 with single lesions, 21%; P<0.01). Single infarctions occurred frequently in the territory of the ruptured aneurysm (22/28 patients; 79%), whereas multiple infarctions were often distant to the site of rupture (21/28 cases, 75%). History of diabetes (P=0.05), early hydrocephalus (P=0.05), and requirement of external ventricular drainage (P=0.02) were associated with the occurrence of multiple infarctions on univariate analysis. On multivariable analysis, this association only remained significant for the requirement of external ventricular drainage. CONCLUSIONS: The 2 most common patterns of delayed cerebral ischemia after aneurysmal SAH are single cortical infarction, typically near the ruptured aneurysm, and multiple widespread lesions including subcortical locations and often unrelated to the site of aneurysm rupture. These 2 patterns may represent different pathophysiological mechanisms or different degrees of severity of the same vascular process. PMID- 15831837 TI - Molecular determinants for the interaction of the valvulopathic anorexigen norfenfluramine with the 5-HT2B receptor. AB - S-(+)-Norfenfluramine (SNF)-an active metabolite of the now-banned anorexigen fenfluramine-has been implicated in the drug's appetite-suppressing actions and its life-threatening cardiovascular side effects. SNF reduces appetite through serotonin 5-HT(2C) receptor activation; it causes cardiopulmonary side effects through 5-HT(2B) receptor activation. Thus, we attempted to identify molecular determinants of SNF binding to 5-HT(2B) receptors distinct from those underlying SNF-5-HT(2C/2A) receptor interactions. Mutagenesis implicated Val2.53 in SNF binding to 5-HT(2B) receptors. Ligand docking simulations suggested both Val2.53 gamma-methyl groups form stabilizing van der Waals' (vdW) interactions with the alpha-methyl group of SNF. A V2.53L mutation induced a 17-fold decrease in affinity; molecular dynamics (MD) simulations suggested that this decrease resulted from the loss of one 2.53-alpha-methyl group vdW interaction. Supporting this, 1) the binding of norfenfluramine (NF) analogs lacking an S-(+) alpha methyl group (RNF and alpha-desmethyl-NF) was less sensitive to the V2.53L mutation, and 2) a V2.53A mutation decreased SNF affinity 190-fold, but decreased RNF and alpha-desmethyl-NF affinities only 16- and 45-fold, respectively. We next addressed whether the alpha-methyl group of SNF contributes to 5-HT(2C/2A) receptor affinity. Removal of the alpha-methyl group (RNF and alpha-desmethyl NF), which reduced 5-HT(2B) receptor binding 3-fold, did not affect 5-HT(2C/2A) receptor binding. An alpha-ethyl substituent (alpha-ethyl-NF), which decreased 5 HT(2B) receptor affinity 46-fold, reduced 5-HT(2C) and 5-HT(2A) receptor binding by 14- and 5-fold, respectively. Finally, we determined that residue 2.53 affects SNF potency and efficacy at 5-HT(2B) receptors but not at 5-HT(2C) and 5-HT(2A) receptors. In conclusion, vdW interactions between residue 2.53 and the alpha methyl group of SNF contribute to the ligand's 5-HT(2) receptor subtype-selective pharmacology. PMID- 15831838 TI - Changes in airway dimensions on computed tomography scans of children with cystic fibrosis. AB - RATIONALE: In cystic fibrosis (CF), chronic bacterial infection and inflammation lead to progressive airway wall thickening and lumen dilatation. OBJECTIVES: To quantify airway wall thickening and lumen dilatation in children with CF over a 2 year interval. METHODS: Children with CF (n = 23) who had two computed tomography (CT) scans (CT(cf1) and CT(cf2)) combined with pulmonary function tests (PFTs), with a 2-year interval between measurements, were compared with control subjects (n = 21) who had one CT (CT(controls)). On cross-sectional cut airway-artery pairs, airway wall area (WA), airway lumen area (LA) and perimeter, and arterial area (AA) were quantified. LA/AA (= marker of bronchiectasis), airway wall thickness (AWT), and WA/AA (= markers of wall thickness) were calculated. CT scans were scored using four different scoring systems. PFTs were expressed as percent predicted. RESULTS: Airway WA-to-AA ratio was 1.45 (p < 0.001) and airway LA-to-AA ratio was 1.92 times higher (p < 0.001) in children with CF compared with age-matched control subjects. LA/AA and WA/AA remained unchanged from CT(cf1) to CT(cf2) and did not increase with age. AWT as a function of airway size increased from CT(cf1) to CT(cf2) by 2% (0.03 mm; p = 0.02). The change in AWT was inversely related to the change in forced expiratory flow between 25 and 75% of expiratory VC (p = 0.002). CONCLUSIONS: In CF, quantitative measurements of airways on CT scans show an increased ratio between airway LA and AA and progressive airway wall thickening. Scoring systems show progression of bronchiectasis but unchanged AWT. PFTs remained stable. PMID- 15831839 TI - The effect of infective exacerbations on sleep and neurobehavioral function in cystic fibrosis. AB - RATIONALE: Adults with cystic fibrosis (CF) are susceptible to hypoxemia, hypercapnia, arousal from sleep, and neurobehavioral impairment. OBJECTIVES: We hypothesized that pulmonary exacerbations would adversely affect sleep and neurobehavioral performance. METHODS: Patients with exacerbations (cases) had sleep studies and neurobehavioral testing before and after inpatient intravenous therapy. Adults with stable CF underwent the same testing procedures (control subjects). MEASUREMENTS AND MAIN RESULTS: When clinically stable, cases and control subjects had similar lung function, intelligence, and body mass index. Among cases, treatment of an exacerbation improved lung function, quality of life, mood, sleepiness, and activation. Cases spent more time awake after sleep onset (p = 0.02), less time in REM sleep (p = 0.03), and were more hypoxemic than control subjects when unwell. The severity of hypoxemia correlated with lung function. On admission, cases had slower throughput than control subjects in the serial addition and subtraction task (cases, 16 +/- 4, vs. control subjects, 17 +/- 3; F[1, 36] = 5.15, p = 0.03) and a slower response time on the digit symbol substitution task (F[1, 36] = 11.91, p = 0.001), which persisted after treatment (F[1, 36] = 8.48, p = 0.006). Cases experienced significant improvements in sleep efficiency, amount of REM sleep, and hypoxia with treatment. Their performance in the serial addition and subtraction task, psychomotor vigilance task, and simulated driving task also improved with treatment. Sex modified the effect of an exacerbation on some aspects of performance. CONCLUSIONS: Exacerbations of lung disease in adults with CF adversely affect sleep and tests of neurobehavioral performance regardless of underlying disease severity. The implications for performance in daily life need further evaluation because patients often delay admission to hospital to fulfill study or work commitments. PMID- 15831840 TI - Rate of reinfection tuberculosis after successful treatment is higher than rate of new tuberculosis. AB - RATIONALE: In a high-tuberculosis (TB) incidence area of Cape Town, South Africa, there is a very high rate of unexplained recurrent TB. The incidence of new bacteriologically confirmed disease in the area is 313 per 100,000 individuals. OBJECTIVE: To estimate the rate of recurrent TB attributable to reinfection after successful treatment. METHODS: All patients with reported TB in the area between 1993 and 1998 were followed up to 2001 for disease needing retreatment (recurrences). Patients who were multi-drug-resistant or who had treatment failure, were transferred, or died during treatment were excluded. Analysis was restricted to patients for whom DNA fingerprinting of their Mycobacterium tuberculosis isolates was obtained. Reinfection TB was defined as a recurrent TB episode in which the strains of the separate episodes differed by more than four bands. MEASUREMENTS AND MAIN RESULTS: 612 of 897 (68%) patients had a DNA fingerprint available at enrollment. Median duration of follow-up was 5.2 years. Recurrent TB occurred in 108 of 612 (18%) patients, of whom 61 of 447 (14%) experienced recurrence after successful treatment, and 47 of 165 (28%) experience recurrence after default. Of the 108 patients with recurrent TB, 68 (63%) had a DNA fingerprint in the second episode. Among these patients, 24 of 31 (77%) recurrences after successful treatment and 4 of 37 (11%) recurrences after default were attributable to reinfection. The reinfection disease rate after successful treatment was estimated at 2.2 per 100 person-years. CONCLUSIONS: The age-adjusted incidence rate of TB attributable to reinfection after successful treatment was four times that of new TB. People who had TB once are at a strongly increased risk of developing TB when reinfected. PMID- 15831841 TI - Short-term cigarette smoke exposure enhances allergic airway inflammation in mice. AB - RATIONALE: Epidemiologic studies suggest that tobacco smoke contributes to the prevalence and occurrence of exacerbations in asthma. The effect of active smoking in adolescents with atopy is poorly understood. OBJECTIVES: We developed an experimental model to investigate the influence of smoking on antigen-induced airway inflammation and airway responsiveness in mice that were previously sensitized. METHODS: Ovalbumin (OVA)-sensitized BALB/c mice were exposed to air or mainstream smoke (5 days/week) and to phosphate-buffered saline (PBS) or OVA aerosol (3 times/week) for 2 weeks (n = 8 for each group). RESULTS: Airway responsiveness to intravenously injected carbachol was increased (p < 0.05) in smoke- and OVA-exposed mice compared with all other groups. There was an additive effect of smoke and OVA exposure on total cell numbers, macrophages, and dendritic cells in bronchoalveolar lavage fluid and on CD4+ and CD8+ T lymphocytes and dendritic cells in lung tissue (p < 0.05 compared with mice exposed to smoke and PBS and to mice exposed to air and OVA). Concurrent smoke and OVA exposure augmented OVA-specific IgE in serum compared with air and OVA exposure. In lavage fluid supernatant, eotaxin was increased in air- and OVA exposed mice. The further increase observed in the group exposed to both OVA and cigarette smoke came close to formal significance (p = 0.06). Thymus- and activation-regulated chemokine was augmented in mice exposed to either smoke or OVA, without additional effect. CONCLUSIONS: Our data indicate that acute concurrent exposure to allergen and mainstream cigarette smoke enhances airway inflammation and airway responsiveness in previously sensitized mice. PMID- 15831829 TI - Melioidosis: epidemiology, pathophysiology, and management. AB - Melioidosis, caused by the gram-negative saprophyte Burkholderia pseudomallei, is a disease of public health importance in southeast Asia and northern Australia that is associated with high case-fatality rates in animals and humans. It has the potential for epidemic spread to areas where it is not endemic, and sporadic case reports elsewhere in the world suggest that as-yet-unrecognized foci of infection may exist. Environmental determinants of this infection, apart from a close association with rainfall, are yet to be elucidated. The sequencing of the genome of a strain of B. pseudomallei has recently been completed and will help in the further identification of virulence factors. The presence of specific risk factors for infection, such as diabetes, suggests that functional neutrophil defects are important in the pathogenesis of melioidosis; other studies have defined virulence factors (including a type III secretion system) that allow evasion of killing mechanisms by phagocytes. There is a possible role for cell mediated immunity, but repeated environmental exposure does not elicit protective humoral or cellular immunity. A vaccine is under development, but economic constraints may make vaccination an unrealistic option for many regions of endemicity. Disease manifestations are protean, and no inexpensive, practical, and accurate rapid diagnostic tests are commercially available; diagnosis relies on culture of the organism. Despite the introduction of ceftazidime- and carbapenem-based intravenous treatments, melioidosis is still associated with a significant mortality attributable to severe sepsis and its complications. A long course of oral eradication therapy is required to prevent relapse. Studies exploring the role of preventative measures, earlier clinical identification, and better management of severe sepsis are required to reduce the burden of this disease. PMID- 15831842 TI - Severe pulmonary hypertension and chronic obstructive pulmonary disease. AB - RATIONALE: Severe pulmonary hypertension occurs occasionally in patients with chronic obstructive pulmonary disease (COPD), but no detailed description of these patients is available. OBJECTIVES: To identify and characterize patients with COPD and severe pulmonary hypertension. METHODS: Retrospective study of 27 patients with COPD with severe pulmonary hypertension (pulmonary artery mean pressure [Ppa], > or = 40 mm Hg) among 998 patients who underwent right heart catheterization between 1990 and 2002 as part of a workup for chronic respiratory failure during a period of disease stability. RESULTS: Of the 27 patients, 16 had another disease capable of causing pulmonary hypertension. The remaining 11 (11 of 998, 1.1%) patients had COPD as the only cause of pulmonary hypertension, with a median Ppa of 48 mm Hg (interquartile range, 46-50). They had an unusual pattern of cardiopulmonary abnormalities with mild to moderate airway obstruction, severe hypoxemia, hypocapnia, and a very low diffusing capacity for carbon monoxide (p < 0.01 compared with a control group of patients with COPD). Exertional dyspnea was more severe (p < 0.01) and survival was shorter (p = 0.0026) than in the control subjects. CONCLUSIONS: Severe pulmonary hypertension is uncommon in patients with COPD. When it occurs, another cause must be sought. COPD with severe pulmonary hypertension and no other possible cause shares features with pulmonary vascular diseases, such as idiopathic pulmonary hypertension. PMID- 15831843 TI - Array lessons from the heart: focus on the genome and transcriptome of cardiomyopathies. AB - Our understanding of the cardiovascular system has evolved through the years by extensive studies emphasizing the identification of the molecular and physiological mechanisms involved in its normal function and disease pathogenesis. Major discoveries have been made along the way. However, the majority of this work has focused on specific genes or pathways rather than integrative approaches. In cardiomyopathies alone, over 30 different loci have shown mutations with varying inheritance patterns, yet mostly coding for structural proteins. The emergence of microarrays in the early 1990s paved the way to a new era of cardiovascular research. Microarrays dramatically accelerated the rhythm of discoveries by giving us the ability to simultaneously study thousands of genes in a single experiment. In the field of cardiovascular research, microarrays are having a significant contribution, with the majority of work focusing on end-stage cardiomyopathies that lead to heart failure. Novel molecular mechanisms have been identified, known pathways are seen under new light, disease subgroups begin to emerge, and the effects of various drugs are molecularly dissected. This cross-study data comparison concludes that consistent energy metabolism gene expression changes occur across dilated, hypertrophic, and ischemic cardiomyopathies, while Ca2+ homeostasis changes are prominent in the first two cardiomyopathies, and structural gene expression changes accompany mostly the dilated form. Gene expression changes are further correlated to disease genetics. The future of microarrays in the cardiomyopathy field is discussed with an emphasis on optimum experimental design and on applications in diagnosis, prognosis, and drug discovery. PMID- 15831844 TI - Inactivation of CD11b in a mouse transgenic model protects against sepsis-induced lung PMN infiltration and vascular injury. AB - To inactivate chronically the beta2-integrin CD11b (Mac-1), we made a transgenic model in mice in which we expressed the CD11b antagonist polypeptide neutrophil inhibitory factor (NIF). Using these mice, we determined the in vivo effects of CD11b inactivation on polymorphonuclear leukocyte (PMN) function and acute lung injury (ALI) induced by Escherichia coli septicemia. In wild-type PMNs, CD11b expression was induced within 1 h after E. coli challenge, whereas this response was significantly reduced in NIF(+/+) PMNs. Coimmunoprecipitation studies showed that NIF associated with CD11b in NIF(+/+) PMNs. To validate the effectiveness of CD11b blockade, we compared PMN function in NIF(+/+) and Mac-1-deficient (Mac-1( /-)) mice. Adhesion of both Mac-1(-/-) and NIF(+/+) PMNs to endothelial cells in response to LPS was reduced in both types of PMNs and fully blocked only by the addition of anti-CD11a monoclonal antibody. This finding is indicative of intact CD11a function in the NIF(+/+) PMNs but the blockade of CD11b function. CD11b inactivation in NIF(+/+) mice interfered with lung PMN infiltration induced by E. coli and prevented the increase in lung microvessel permeability and edema formation, with most of the protection seen in the 1-h period after the E. coli. Thus our results demonstrate that CD11b plays a crucial role in mediating lung PMN sequestration and vascular injury in the early phase of gram-negative septicemia. The NIF(+/+) mouse model, in which CD11b is inactivated by binding to NIF, is a potentially useful model for in vivo assessment of the role of PMN CD11b in the mechanism of vascular inflammation. PMID- 15831845 TI - Female sexual dysfunction. PMID- 15831847 TI - The influence of big pharma. PMID- 15831828 TI - Molecular pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies. AB - Bordetella respiratory infections are common in people (B. pertussis) and in animals (B. bronchiseptica). During the last two decades, much has been learned about the virulence determinants, pathogenesis, and immunity of Bordetella. Clinically, the full spectrum of disease due to B. pertussis infection is now understood, and infections in adolescents and adults are recognized as the reservoir for cyclic outbreaks of disease. DTaP vaccines, which are less reactogenic than DTP vaccines, are now in general use in many developed countries, and it is expected that the expansion of their use to adolescents and adults will have a significant impact on reducing pertussis and perhaps decrease the circulation of B. pertussis. Future studies should seek to determine the cause of the unique cough which is associated with Bordetella respiratory infections. It is also hoped that data gathered from molecular Bordetella research will lead to a new generation of DTaP vaccines which provide greater efficacy than is provided by today's vaccines. PMID- 15831848 TI - Reducing mortality in myocardial infarction. PMID- 15831849 TI - Drugs for Alzheimer's disease and related dementias. PMID- 15831850 TI - Human and veterinary medicine. PMID- 15831851 TI - Treatment of postmenopausal osteoporosis. PMID- 15831852 TI - Mother convicted of killing her children is freed on appeal. PMID- 15831853 TI - NHS is failing to reap benefit of new technologies, MPs say. PMID- 15831854 TI - Wet nursing increases risk of HIV infection among babies. PMID- 15831855 TI - Pfizer is asked to suspend sales of painkiller. PMID- 15831856 TI - Scientists develop prototype of more sensitive test for cancer. PMID- 15831857 TI - Some countries will not achieve millennium development goals for health by 2015, WHO says. PMID- 15831859 TI - MPs call for free personal care for elderly people. PMID- 15831860 TI - French doctors go on strike to demand reintroduction of compulsory out of hours work. PMID- 15831861 TI - Attempts to contain Marburg disease epidemic have limited effect. PMID- 15831862 TI - New rules drive off NIH researchers. PMID- 15831867 TI - GPs are dissatisfied with the care they are giving patients with eating disorders. PMID- 15831864 TI - Drug company is reprimanded by UK regulator. PMID- 15831868 TI - UK response to global HIV epidemic is condemned for lack of coordination. PMID- 15831869 TI - US states get healthcare ratings. PMID- 15831870 TI - Canadian doctors suggest acceptable waiting times for care. PMID- 15831872 TI - A kinder, gentler age. PMID- 15831871 TI - EU to tackle issue of patient safety. PMID- 15831873 TI - "Frequent flier" patients. PMID- 15831874 TI - The patient's journey: multiple sclerosis. PMID- 15831875 TI - Postoperative hypoxia in a woman with Down's syndrome: case progression. PMID- 15831876 TI - A life threatening complication after colonoscopy. PMID- 15831877 TI - Suicide and deliberate self harm in young people. PMID- 15831878 TI - Reader's guide to critical appraisal of cohort studies: 1. Role and design. PMID- 15831879 TI - Improving diet and physical activity: 12 lessons from controlling tobacco smoking. PMID- 15831880 TI - Young people's health: the need for action. PMID- 15831881 TI - Switching prescription drugs to over the counter: consumers may benefit financially in examples from the US. PMID- 15831885 TI - Switching prescription drugs to over the counter: willingness to buy statin over the counter is not related to risk of heart disease. PMID- 15831886 TI - Switching prescription drugs to over the counter: chicken and egg scenario is possible. PMID- 15831887 TI - Mapping choice in the NHS: analysis is only as good as data. PMID- 15831888 TI - Direct to consumer marketing: advertising directly to consumers may affect patients' safety. PMID- 15831889 TI - Direct to consumer marketing: switch to over the counter can become indirect advertising directly to consumers. PMID- 15831890 TI - Switching prescription drugs to over the counter: over the counter licensing is linked to direct to consumer advertising. PMID- 15831892 TI - Rising healthcare costs, equity, and electoral success constitute trio of tensions. PMID- 15831893 TI - UK guidelines for the management of acute pancreatitis. PMID- 15831894 TI - Brain imaging and its implications for studying centrally targeted treatments in irritable bowel syndrome: a primer for gastroenterologists. AB - Research into brain-gut interactions, and the use of brain imaging, as potential investigative tools for functional gastrointestinal disorders, such as irritable bowel syndrome, is a promising new area. Studies are beginning to identify the structure and function of regions of the brain and their relationships to pain perception, stress, and other psychosocial variables. These imaging modalities may also have diagnostic potential, and perhaps even therapeutic applications, particularly with regard to understanding the benefit of centrally targeted modalities such as antidepressants and psychological treatments. PMID- 15831895 TI - Clinical epidemiology--how important now? PMID- 15831896 TI - Increased deoxycholic acid absorption and gall stones in acromegalic patients treated with octreotide: more evidence for a connection between slow transit constipation and gall stones. PMID- 15831897 TI - Reversal of DNA hypomethylation by folic acid supplements: possible role in colorectal cancer prevention. PMID- 15831898 TI - Autoimmune pancreatitis--also a Western disease. PMID- 15831899 TI - Roles of virD4 and cagG genes in the cag pathogenicity island of Helicobacter pylori using a Mongolian gerbil model. AB - BACKGROUND AND AIMS: The roles of the virD4 and the cagG genes in the cag pathogenicity island of Helicobacter pylori for gastroduodenal pathogenesis are unclear and their roles in vivo have not been examined. METHODS: Seven week old male Mongolian gerbils were inoculated with the wild type H pylori TN2GF4, its isogenic virD4, or cagG mutants. Animals were sacrificed at 4, 12, and 24 weeks after inoculation. Gastric inflammation and H pylori density were evaluated by histology, inflammatory response (as measured by interleukin (IL)-1beta mRNA levels), proliferative activity (as assessed by 5'-bromo-2'deoxyuridine labelling indices), and host systemic reaction (as measured by anti-H pylori IgG antibody). RESULTS: Degree of gastric inflammation, proliferative activity, and mucosal IL 1beta mRNA levels remained low throughout the first 12 weeks in gerbils infected with the virD4 mutants. Degree of gastric inflammation and proliferative activity increased at 24 weeks with the virD4 mutants reaching levels comparative with those seen at four weeks with the wild-type strains. Mucosal IL-1beta mRNA levels were also increased at 24 weeks with the virD4 mutants and levels at 24 weeks were similar between the wild-type and virD4 mutants. In contrast, gerbils infected with the cagG mutants had reduced ability to colonise gerbils, and no or little gastric inflammation or proliferative activity was observed. CONCLUSIONS: Loss of the virD4 gene temporally retarded but did not abrogate gastric inflammation. Loss of the cagG gene abolished gastric inflammation partially via reduced ability to colonise gerbils. Unknown factors related to the type IV secretion system other than CagA may influence gastric inflammation. PMID- 15831900 TI - Insulin-like growth factor I receptor blockade enhances chemotherapy and radiation responses and inhibits tumour growth in human gastric cancer xenografts. AB - BACKGROUND AND AIMS: Insulin-like growth factor (IGF) I receptor (IGF-Ir) signalling is required for carcinogenicity and proliferation of many tumours but this pathway has not been studied in detail in gastric cancer. We have previously shown successful therapy for colorectal, pancreatic, and lung cancer using recombinant adenoviruses expressing dominant negative (dn) IGF-Ir. In this study, we sought to better dissect the role of IGF-Ir on progression of gastric cancer and determine whether IGF-Ir targeted adenoviruses represent potentially effective therapeutics for human gastric cancer. METHODS: We assessed the effect of IGF-Ir ligands on proliferation and survival in gastric cancer cells in culture. Then, recombinant adenoviruses expressing truncated IGF-Ir (482 and 950 amino acids long, IGF-Ir/dn) that function as dn inhibitors were studied in the treatment of human gastric cancer xenografts. We characterised the effects of IGF Ir/dn on signalling blockade, growth, apoptosis induction, and in vivo therapeutic efficacy. RESULTS: IGF-Ir signalling promoted tumour growth and survival in gastric cancer. IGF-Ir/dn expression suppressed tumorigenicity both in vitro and in vivo and upregulated stressor induced apoptosis. IGF-Ir/dn blocked Akt-1 activation induced by IGF-I, IGF-II, and des(1-3)IGF-I, but not by insulin. IGF-Ir/dn expression increased radiation and chemotherapy induced apoptosis and the combination of IGF-Ir/dn and chemotherapy was very effective against tumours in mice. In an intraperitoneal model, IGF-Ir/dn therapy also suppressed peritoneal dissemination. CONCLUSIONS: IGF-Ir is involved in the regulation of survival and cell growth in human gastric cancer and may be a good molecular therapeutic target. Adenovirus-IGF-Ir/dn may thus have therapeutic use in gastric cancer. PMID- 15831901 TI - Amitriptyline reduces rectal pain related activation of the anterior cingulate cortex in patients with irritable bowel syndrome. AB - BACKGROUND AND AIMS: Irritable bowel syndrome (IBS) is a disorder of intestinal hypersensitivity and altered motility, exacerbated by stress. Functional magnetic resonance imaging (fMRI) during painful rectal distension in IBS has demonstrated greater activation of the anterior cingulate cortex (ACC), an area relevant to pain and emotions. Tricyclic antidepressants are effective for IBS. The aim of this study was to determine if low dose amitriptyline reduces ACC activation during painful rectal distension in IBS to confer clinical benefits. Secondary aims were to identify other brain regions altered by amitriptyline, and to determine if reductions in cerebral activation are greater during mental stress. METHODS: Nineteen women with painful IBS were randomised to amitriptyline 50 mg or placebo for one month and then crossed over to the alternate treatment after washout. Cerebral activation during rectal distension was compared between placebo and amitriptyline groups by fMRI. Distensions were performed alternately during auditory stress and relaxing music. RESULTS: Rectal pain induced significant activation of the perigenual ACC, right insula, and right prefrontal cortex. Amitriptyline was associated with reduced pain related cerebral activations in the perigenual ACC and the left posterior parietal cortex, but only during stress. CONCLUSIONS: The tricyclic antidepressant amitriptyline reduces brain activation during pain in the perigenual (limbic) anterior cingulated cortex and parietal association cortex. These reductions are only seen during stress. Amitriptyline is likely to work in the central nervous system rather than peripherally to blunt pain and other symptoms exacerbated by stress in IBS. PMID- 15831902 TI - Role of cyclooxygenases 1 and 2 in the modulation of neuromuscular functions in the distal colon of humans and mice. AB - BACKGROUND: Cyclooxygenase isoforms (COX-1, COX-2) may exert differential regulatory actions on enteric motor functions under normal or pathological conditions. AIMS: To examine the occurrence and functions of COX-1 and COX-2 in the neuromuscular compartment of normal distal colon using human and murine tissue. METHODS: Gene expression (human, mouse), protein expression (human), gene deletion (mouse), and the effects of dual and isoform specific COX inhibitors on in vitro motility (human, mouse) were investigated. RESULTS: Reverse transcription-polymerase chain reaction (RT-PCR) showed mRNA expression of COX-1 and COX-2 in human and wild-type mouse colonic muscle whereas only COX-2 or COX-1 was detected in COX-1 or COX-2 knockout animals. Immunohistochemistry localised both isoforms in neurones of myenteric ganglia, COX-1 in circular layer myocytes, and COX-2 in longitudinal muscle. Indomethacin (COX-1/COX-2 inhibitor), SC-560 (COX-1 inhibitor), or DFU (COX-2 inhibitor) enhanced atropine sensitive electrically induced contractions of human longitudinal muscle. The most prominent actions were recorded with indomethacin or SC-560 plus DFU. These results were confirmed under pharmacological blockade of non-cholinergic nerves. Atropine sensitive contractions evoked by carbachol in the presence of tetrodotoxin were enhanced by indomethacin or DFU but not by SC-560. In wild-type mice, contractile responses to electrical stimulation were enhanced by indomethacin, SC-560, or DFU. SC-560 potentiated electrically induced contractions in COX-2, but not COX-1, knockout mice. In contrast, DFU enhanced the contractions elicited by electrical stimuli in COX-1, but not in COX-2, knockout mice. CONCLUSIONS: These results indicate that COX-1 and COX-2 are expressed in the neuromuscular compartment of normal human colon where they modulate cholinergic excitatory control of colonic motility at prejunctional and postjunctional sites, respectively. PMID- 15831903 TI - An unusual case of terminal ileal stricture. PMID- 15831904 TI - Risk of haematopoietic cancer in patients with inflammatory bowel disease. AB - BACKGROUND AND AIMS: Several chronic inflammatory conditions are associated with an increased risk of lymphoma. Whether this applies to inflammatory bowel disease (IBD) is still unclear but of paramount interest, particularly in the safety evaluation of newer immunosuppressive drugs. Reports also indicate a possible increase in the risk of leukaemia in IBD. We therefore assessed the risk of haematopoietic cancers in a large cohort of patients with IBD. SUBJECTS AND METHODS: We performed a population based cohort study using prospectively recorded data, including 47 679 Swedish patients with Crohn's disease (CD) or ulcerative colitis (UC) assembled from regional cohorts of IBD from 1955 to 1990 (n = 8028) and from the Inpatient Register of 1964-2000 (n = 45 060), with follow up until 2001. Relative risks were expressed as standardised incidence ratios (SIR). RESULTS: Overall, we observed 264 haematopoietic cancers during follow up, which corresponded to a borderline significant 20% increased risk in both UC and CD. In UC, lymphomas occurred as expected (SIR 1.0, n = 87) but myeloid leukaemia occurred significantly more often than expected (SIR 1.8, n = 32). In CD, there was a borderline significant increased lymphoma risk (SIR 1.3, n = 65), essentially confined to the first years of follow up. Proxy markers of disease activity had little impact on lymphoma risk. CONCLUSION: On average, patients with IBD have a marginally increased risk of haematopoietic cancer. In UC, this is accounted for by an excess of myeloid leukaemia. In CD, a modest short term increase in the risk of lymphoma of unknown significance cannot be excluded but any long term risk increase seems unlikely. PMID- 15831906 TI - How to evaluate a gastric submucosal tumour in a patient with haematemesis? PMID- 15831905 TI - Activation of RegIIIbeta/gamma and interferon gamma expression in the intestinal tract of SCID mice: an innate response to bacterial colonisation of the gut. AB - BACKGROUND AND AIMS: The mechanisms by which commensal bacteria provoke intestinal inflammation in animal models of inflammatory bowel disease (IBD) remain incompletely defined, leading to increasing interest in the innate immune response of the colonic mucosa to bacterial colonisation. METHODS: Using gene expression profiling of colonic RNA from C.B17.SCID germ free mice and those colonised with altered Schaedler's flora, we investigated the innate immune response to bacterial colonisation in vivo. The two most consistently induced gene groups were RegIIIbeta and gamma as well as interferon gamma (IFN-gamma) response genes. RESULTS: Using quantitative reverse transcription-polymerase chain reaction, we showed that RegIIIbeta, RegIIIgamma, and IFN-gamma were constitutively expressed in the colon of conventionally housed SCID mice compared with either germ free SCID or conventionally housed BALB/c mice. Induction of these genes was reproduced by chronic monoassociation of germ free SCID mice with either of two separate gut commensal bacterial species-segmented filamentous bacteria and Schaedler's Escherichia coli. The cellular source for IFN-gamma on monoassociation of SCID mice with Schaedler's E coli was localised to a subset of intraepithelial natural killer (IENK) cells that express asialo-GM1. In vivo IFN gamma immunoneutralisation studies failed to demonstrate any alteration in RegIIIbeta or gamma expression. CONCLUSIONS: Thus bacterial colonisation of the colon independently activates two distinct innate immune cell types at the mucosal interface with the colonic lumen, intestinal epithelial cells, and IENK cells, a response that may be regulated by the adaptive immune system. These innate immune responses may play a role in the pathogenesis of colitis in SCID adoptive transfer models in mice and possibly in patients with IBD. PMID- 15831907 TI - Octreotide induced prolongation of colonic transit increases faecal anaerobic bacteria, bile acid metabolising enzymes, and serum deoxycholic acid in patients with acromegaly. AB - BACKGROUND: Acromegalic patients have slow colonic transit, increased rates of deoxycholic acid formation, and an increased prevalence of cholesterol gall stones, especially during long term octreotide treatment. However, the effects of this prolonged large bowel transit time on the numbers of faecal anaerobes and the activities of the enzyme systems which biotransform conjugated cholic acid into unconjugated deoxycholic acid (cholylglycine hydrolase and 7alpha dehydroxylase) are unknown. METHODS: Therefore, in 10 non-acromegalic controls, 11 acromegalic patients not treated with octreotide, and 11 acromegalics on long term (8-48 months) octreotide (100-200 mug three times daily subcutaneously), we measured large bowel transit time and, in freshly voided faeces, the activities of the two bile acid metabolising enzymes, and related the results to the proportion of deoxycholic acid in fasting serum. Moreover, in patients with acromegaly, we measured quantitative bacteriology in faeces. RESULTS: Mean large bowel transit time in acromegalics not treated with octreotide (35 (SEM 6.5) hours) was 66% longer than that in non-acromegalic controls (21 (3.1) hours; NS) and became further prolonged during octreotide treatment (48 (6.6) hours; p<0.001). These octreotide induced changes in transit were associated, in acromegalic patients, with more total (15.0 (2.5) v 6.3 (1.3)x10(9) colony forming units (cfu)/g; p<0.05) and Gram positive (6.3 (2.3) v 3.2 (1.0)x10(9) cfu/g; p<0.05) faecal anaerobes. Mean faecal cholylglycine hydrolase activity in the long term octreotide group (22.0 (6.0)x10(-2) U/mg protein) was 138% greater than that in non-acromegalic controls (12.0 (6.0)x10(-2); p<0.01). Similarly, mean 7alpha-dehydroxylase activity in octreotide treated acromegalics (11.1 (1.18)x10(-4) U/mg protein) was 78% greater than that in patients not receiving long term octreotide (6.3 (0.5)x10(-4); p<0.001). The mean proportion of deoxycholic acid in fasting serum also increased from 18.0 (2.88)% in the untreated group to 29.6 (2.3)% during long term octreotide (p<0.05). There were significant linear relationships between large bowel transit time and: (i) faecal 7alpha-dehydroxylase activity; and (ii) the proportion of deoxycholic acid in fasting serum and between 7alpha-dehydroxylase activity and the proportion of deoxycholic acid in serum.Summary/interpretation: These data suggest that increased deoxycholic acid formation seen in acromegalics during octreotide treatment is due not only to the greater numbers of faecal anaerobes but also to increased activity of the rate limiting enzyme pathway (7alpha-dehydroxylation) converting cholic acid to deoxycholic acid. PMID- 15831908 TI - Association between family history and mismatch repair in colorectal cancer. AB - BACKGROUND AND AIMS: Germline mutations in mismatch repair (MMR) genes cause a greatly increased risk of cancer of the gastrointestinal and female reproductive tracts (hereditary non-polyposis colorectal cancer (HNPCC)). Loss of MMR expression is common in colorectal cancer (CRC) overall. Such loss is assumed to be acquired predominantly, although a population of CRC cases will include individuals with unrecognised MMR mutations. This study examines the association between MMR gene expression and family history of cancer among the CRC population. METHODS: Individuals with CRC were identified from two well characterised populations: (1) consecutive hospital patients (n = 644) and (2) a population based cases series (n = 249). CRC was examined for expression of hMLH1 and hMSH2 using immunohistochemistry, and expression was related to family history using logistic regression. RESULTS: hMLH1 and hMSH2 expression was assessed in 732 CRCs with 8% showing loss of expression. No association was seen overall for hMLH1 or hMSH2 expression and family history of CRC. Loss of hMSH2 was predicted by family history of extracolonic cancer (odds ratio (OR) 5.78 (95% confidence interval (CI) 0.95-35.18)) and family history suggestive of HNPCC (OR 27.84 (95% CI 4.37-177.56)). Loss of hMLH1 was not predicted by family history of extracolonic cancer or a family history suggestive of HNPCC but was for a family history of at least two affected relatives (OR 4.88 (95% CI 1.25-19.03)). CONCLUSIONS: Individuals with hMSH2 deficient CRC in the general population exhibit a family history and other characteristics suggestive of HNPCC, and may carry germline MMR mutations. Loss of hMLH1 is only associated with a strong family history of extracolonic cancer at older ages, suggesting a novel mechanism of susceptibility. PMID- 15831909 TI - The C/C-13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population. AB - BACKGROUND AND AIMS: The role of nutrition in the pathogenesis of colorectal cancer is not fully understood. Milk products are an essential part of human nutrition in Western countries. Absorption of lactose, the main sugar of milk, is regulated by the activity of the lactase enzyme in the gut wall. The activity of lactase is genetically determined and is associated with a C/T single nucleotide polymorphism residing 13910 bp upstream of the lactase coding sequence. Here we have studied the relationship between the C/T(-13910) polymorphism and colorectal cancer in Finnish, British, and Spanish populations. PATIENTS AND METHODS: A total of 2766 subjects, including 963 Finnish, 283 British, and 163 Spanish subjects with colorectal cancer, and 773 Finnish, 363 British, and 221 Spanish control subjects, were genotyped for the C/T(-13910) variant by polymerase chain reaction minisequencing. RESULTS: The C/C(-13910) genotype, which is a robust molecular marker of low lactase activity (lactase non-persistence), was found to significantly associate with the risk of colorectal cancer (p = 0.015) in the Finnish subjects, with an odds ratio of 1.40 (95% confidence interval 1.07-1.85). No association was found with site, histology, or stage of the tumour. No significant risk was detected in the British or Spanish populations. CONCLUSION: Low lactase enzyme activity, defined by genotyping of the C/T(-13910) variant, may increase the risk of colorectal cancer. Further studies are warranted to investigate the role of milk and other dairy products in the pathogenesis of colon cancer in different populations. PMID- 15831910 TI - Effect of folic acid supplementation on genomic DNA methylation in patients with colorectal adenoma. AB - BACKGROUND AND AIMS: A low dietary folate intake can cause genomic DNA hypomethylation and may increase the risk of colorectal neoplasia. The hypothesis that folic acid supplementation increases DNA methylation in leucocytes and colorectal mucosa was tested in 31 patients with histologically confirmed colorectal adenoma using a randomised, double blind, placebo controlled, parallel design. METHODS: Subjects were randomised to receive either 400 microg/day folic acid supplement (n = 15) or placebo (n = 16) for 10 weeks. Genomic DNA methylation, serum and erythrocyte folate, and plasma homocysteine concentrations were measured at baseline and post intervention. RESULTS: Folic acid supplementation increased serum and erythrocyte folate concentrations by 81% (95% confidence interval (CI) 57-104%; p<0.001 v placebo) and 57% (95% CI 40-74%; p<0.001 v placebo), respectively, and decreased plasma homocysteine concentration by 12% (95% CI 4-20%; p = 0.01 v placebo). Folic acid supplementation resulted in increases in DNA methylation of 31% (95% CI 16-47%; p = 0.05 v placebo) in leucocytes and 25% (95% CI 11-39%; p = 0.09 v placebo) in colonic mucosa. CONCLUSIONS: These results suggest that DNA hypomethylation can be reversed by physiological intakes of folic acid. PMID- 15831911 TI - Increased microvascular blood content is an early event in colon carcinogenesis. AB - BACKGROUND: Increased premalignant epithelial microvascular blood content is a common theme in neoplastic transformation; however, demonstration of this phenomenon in colon carcinogenesis has been stymied by methodological limitations. Our group has recently developed a novel optics technology, four dimensional elastic light scattering fingerprinting (4D-ELF), which allows examination of the colonic mucosal architecture with unprecedented accuracy. In this study, we utilised 4D-ELF to probe the preneoplastic colonic microvasculature. METHODS: Colonic mucosal blood content was assessed by 4D-ELF at serial preneoplastic time points from azoxymethane (AOM) treated Fisher 344 rats and age matched control animals. We also examined the pretumorigenic intestinal mucosa of the MIN mouse, and compared with wild-type mice. Finally, in a pilot study, we examined superficial blood content from the endoscopically normal mid transverse colon in 37 patients undergoing screening colonoscopy. RESULTS: In the AOM treated rat model, augmentation of superficial mucosal and total mucosal/superficial submucosal blood supply preceded the appearance of aberrant crypt foci (ACF) and temporally and spatially correlated with future ACF occurrence. These findings were replicated in MIN mice. The 4D-ELF based results were corroborated with immunoblot analysis for haemoglobin on mucosal scrapings from AOM treated rats. Moreover, 4D-ELF analysis of normal human colonic mucosa indicated that there was a threefold increase in superficial blood in patients who harboured advanced adenomas. CONCLUSION: We report, for the first time, that blood content is increased in the colonic microvasculature at the earliest stages of colon carcinogenesis. These findings may provide novel insights into early biological events in colorectal carcinogenesis and have potential applicability for screening. PMID- 15831912 TI - Impaired CD95 expression predisposes for recurrence in curatively resected colon carcinoma: clinical evidence for immunoselection and CD95L mediated control of minimal residual disease. AB - BACKGROUND: Loss of CD95 expression in tumour cells occurs frequently in colon carcinoma and may be associated with disease progression. On the other hand, neo expression of CD95L in tumour cells may contribute to immune evasion. AIMS: We aimed at further exploring the functional role and prognostic significance of the CD95/CD95L death inducing system in colon carcinomas. PATIENTS AND METHODS: CD95 and CD95L expression was examined by immunohistochemistry in 128 R0 resected UICC (International Union against Cancer) stage II/III colon carcinomas and correlated with disease free survival. RESULTS: CD95 expression in tumour cells was observed in only 30 carcinomas (23.4%) whereas the others had at least a minor subpopulation of CD95 negative cells. Loss of CD95 in tumour cells was related to adverse prognosis in uni- and multivariate analysis (p = 0.046 and p = 0.036, respectively). Tumour infiltrating lymphocytes (TIL) were the major source of CD95L in colon carcinomas. CD95L+TIL were present in 83% of cases whereas CD95L was found in tumour cells in only 12% of cases. Moreover, a high rate of CD95L+TIL correlated with prolonged disease free survival in patients with UICC stage II (p = 0.05) but not in those with stage III. CONCLUSIONS: Loss of CD95 in tumour cells may be an independent prognostic factor in colon carcinomas. The CD95L counterattack is not a relevant feature in colon carcinoma but CD95L+TIL may contribute to tumour control in the early stages of the disease, exerting a concurrent selection pressure in the direction of CD95 abrogation/resistance. PMID- 15831913 TI - Placenta growth factor expression is correlated with survival of patients with colorectal cancer. AB - BACKGROUND: Overexpression of vascular endothelial growth factor (VEGF) correlates with vascularity, metastasis, and proliferation in colorectal cancer but the role of its homologue, placenta growth factor (PlGF), is unknown. The aim of this study was to evaluate expression and clinical implications of PlGF in colorectal cancer. METHODS: We investigated 74 tumour/non-tumour pairs of colorectal cryosections. Clinical staging was based on the UICC-TNM classification. Expression levels of mRNA for PlGF and VEGF were analysed with quantitative real time reverse transcription-polymerase chain reaction. Proteins were analysed by immunohistochemical staining and enzyme linked immunoabsorbant assay. Analysis of the differences in PlGF and VEGF levels between tumour and non tumour tissues in the same patient were performed by paired t test; differences between localised and advanced disease patients by the Mann-Whitney, chi(2), and Fisher's exact tests and survival curves by the Kaplan-Meier method. RESULTS: Expression levels for both growth factors were significantly higher in tumour than in non-tumour tissues (p/=40 years" was associated with an excess risk of FHF (odds ratio (OR) 2.33 (95% confidence interval (CI) 1.50-3.64)) and death or liver transplantation (OR 4.18 (95% CI 2.17-8.05)). In addition, older age was associated with other risk factors for paracetamol hepatotoxicity such as regular alcohol abuse and late presentation. CONCLUSIONS: Age 40 years or above was identified as a significant independent risk factor for FHF and mortality following paracetamol overdose. Patients aged 40 years or above should be considered as high risk patients, in particular when older age appears in combination with regular alcohol abuse or late presentation. PMID- 15831918 TI - Splanchnic vein thrombosis in candidates for liver transplantation: usefulness of screening and anticoagulation. AB - BACKGROUND AND AIMS: Splanchnic vein thrombosis is a significant source of complications in candidates for liver transplantation. The aims of this study were: (a) to determine the prevalence of and risk factors for splanchnic vein thrombosis in cirrhotic patients awaiting transplantation and (b) to assess the usefulness of anticoagulation. METHODS: A total of 251 cirrhotic patients listed for transplantation were analysed. All underwent systematic screening for thrombosis with Doppler ultrasonography. During the second period of the study, all patients with thrombosis received anticoagulation up to transplantation while during the first period none had received anticoagulation. RESULTS: The incidence of splanchnic vein thrombosis at evaluation was 8.4%. Seventeen additional patients (7.4%) developed de novo thrombosis after evaluation. Independent risk factors for thrombosis were low platelet count (77.4 (36.3) v 111.6 (69.2) 10(9)/l; p = 0.001), a past history of variceal bleeding (47.4% v 29.1%; p = 0.003), and a prolonged interval from listing to transplantation (8.5 (6.8) v 4.8 (4.4) months; p = 0.002). The proportion of partial or complete recanalisation was significantly higher in those who received (8/19) than in those who did not receive (0/10, p = 0.002) anticoagulation. Survival was significantly lower in those who had complete portal vein thrombosis at the time of surgery (p = 0.04). CONCLUSION: These results support a systematic screening for splanchnic vein thrombosis in patients awaiting transplantation. They suggest that in these patients, anticoagulation is safe and has a significant impact on recanalisation as well as prevention of extension of thrombosis. PMID- 15831919 TI - Extrahepatic biliary obstruction after percutaneous tumour ablation for hepatocellular carcinoma: aetiology and successful treatment with endoscopic papillary balloon dilatation. AB - BACKGROUND AND AIMS: Percutaneous tumour ablation (PTA), such as ethanol injection and radiofrequency ablation, is now recognised as a primary treatment for hepatocellular carcinoma (HCC). Although PTA is a relatively safe procedure, it can cause biliary obstruction as a rare complication. As patients with cirrhosis undergoing surgery or endoscopic retrograde cholangiopancreatography/sphincterotomy have a high mortality rate from bleeding, we adopted the use of endoscopic papillary balloon dilatation (EPBD) in these patients and now report the results. We retrospectively analysed the incidence of biliary obstruction after PTA and the efficacy of treatment with EPBD. PATIENTS AND METHODS: A total of 1043 patients with HCC were treated by PTA, of whom 538 were treated with transarterial embolisation with up to eight years of follow up. RESULTS: There were 17 (1.6%) cases of hilar obstruction due to tumour progression and 35 (3.4%) cases of extrahepatic obstruction. Apart from the expected causes of biliary obstruction (haemobilia n = 11, gallstones n = 11, and three miscellaneous causes), we found that 10 patients had obstruction due to biliary casts. This is the first description of biliary casts after percutaneous tumour ablation therapy. Extrahepatic biliary obstruction by procedure related haemobilia occurred within three days of PTA while other causes occurred between 0 and 17 (average 4.9) months. Biliary casts occurred more frequently after ethanol injection than after radiofrequency ablation. EPBD successfully dissipated biliary obstruction in 33 of 35 cases, while two died due to hepatic failure despite successful drainage. CONCLUSIONS: Extrahepatic biliary obstruction is an uncommon complication after PTA for HCC, and can be safely and effectively treated with EPBD, despite impaired liver function. PMID- 15831920 TI - Antibodies to carbonic anhydrase and IgG4 levels in idiopathic chronic pancreatitis: relevance for diagnosis of autoimmune pancreatitis. AB - BACKGROUND: Increased serum antibodies against carbonic anhydrase II (CA-II Ab) or IgG4 levels have been reported in cases of autoimmune chronic pancreatitis (ACP). AIM: To assess the relevance of serum CA-II Ab and IgG4 levels for the diagnosis of ACP in idiopathic CP (ICP) versus alcoholic CP and Sjogren's syndrome (SS). SUBJECTS: This was a multicentre study involving 227 subjects divided into four groups: ICP (n = 54), normal controls (n = 54, paired by age and sex with ICP patients), alcoholic CP (n = 86), and SS (n = 33). METHODS: CA II Ab was measured by ELISA and confirmed by western blotting. A score of easy clinical use with major clinical, morphological, and biochemical parameters for the diagnosis of ACP was applied. RESULTS: The percentage of patients with increased serum CA-II Ab was higher in the ICP group (28%) than in controls (1.9%) and in patients with alcoholic CP (10.5%), but lower than in patients with SS (64%). The proportion with elevated IgG4 levels was higher in the ICP group (15%) compared with controls (1.9%) and SS (0%) but not significantly different from alcoholic CP (8%). Most ICP patients (7/8) with high IgG4 levels exhibited increased CA-II Ab and a compatible ACP score. A definitive diagnosis of ACP by histological analysis was associated with other autoimmune disorders, an increase in both serum IgG4 and CA-II Ab levels, and IgG4 positive plasma cells. CONCLUSIONS: The increase in serum IgG4 levels was strongly associated with elevated CA-II Ab levels, manifestations compatible with ACP, and lymphoplasmacytic infiltration when surgical specimens were available. PMID- 15831922 TI - Epidemiology of gastro-oesophageal reflux disease: a systematic review. AB - A systematic review of the epidemiology of gastro-oesophageal reflux disease (GORD) has been performed, applying strict criteria for quality of studies and the disease definition used. The prevalence and incidence of GORD was estimated from 15 studies which defined GORD as at least weekly heartburn and/or acid regurgitation and met criteria concerning sample size, response rate, and recall period. Data on factors associated with GORD were also evaluated. An approximate prevalence of 10-20% was identified for GORD, defined by at least weekly heartburn and/or acid regurgitation in the Western world while in Asia this was lower, at less than 5%. The incidence in the Western world was approximately 5 per 1000 person years. A number of potential risk factors (for example, an immediate family history and obesity) and comorbidities (for example, respiratory diseases and chest pain) associated with GORD were identified. Data reported in this systematic review can be interpreted with confidence as reflecting the epidemiology of "true" GORD. The disease is more common in the Western world than in Asia, and the low rate of incidence relative to prevalence reflects its chronicity. The small number of studies eligible for inclusion in this review highlights the need for global consensus on a symptom based definition of GORD. PMID- 15831923 TI - Sepsis in cirrhosis: report on the 7th meeting of the International Ascites Club. AB - Sepsis is a systemic inflammatory response to the presence of infection, mediated via the production of many cytokines, including tumour necrosis factor (TNF-), interleukin (IL)-6, and IL-1, which cause changes in the circulation and in the coagulation cascade. There is stagnation of blood flow and poor oxygenation, subclinical coagulopathy with elevated D-dimers, and increased production of superoxide from nitric oxide synthase. All of these changes favour endothelial apoptosis and necrosis as well as increased oxidant stress. Reduced levels of activated protein C, which is normally anti-inflammatory and antiapoptotic, can lead to further tissue injury. Cirrhotic patients are particularly susceptible to bacterial infections because of increased bacterial translocation, possibly related to liver dysfunction and reduced reticuloendothelial function. Sepsis ensues when there is overactivation of pathways involved in the development of the sepsis syndrome, associated with complications such as renal failure, encephalopathy, gastrointestinal bleed, and shock with decreased survival. Thus the treating physician needs to be vigilant in diagnosing and treating bacterial infections in cirrhosis early, in order to prevent the development and downward spiral of the sepsis syndrome. Recent advances in management strategies of infections in cirrhosis have helped to improve the prognosis of these patients. These include the use of prophylactic antibiotics in patients with gastrointestinal bleed to prevent infection and the use of albumin in patients with spontaneous bacterial peritonitis to reduce the incidence of renal impairment. The use of antibiotics has to be judicious, as their indiscriminate use can lead to antibiotic resistance with potentially disastrous consequences. PMID- 15831924 TI - Probiotics in IBD: mucosal and systemic routes of administration may promote similar effects. PMID- 15831925 TI - Efficacy and strategy of pneumatic dilatation in achalasia. PMID- 15831926 TI - Mutations in anionic trypsinogen gene are not associated with tropical calcific pancreatitis. PMID- 15831927 TI - Coexistent chronic idiopathic intestinal pseudo obstruction and inflammatory bowel disease. PMID- 15831928 TI - Elevated plasma protein C levels correlate with the presence of fatty liver (NASH and NAFLD). PMID- 15831929 TI - Inflammatory bowel disease stimulates formation of carcinogenic N-nitroso compounds. PMID- 15831930 TI - Cannabinoid hyperemesis: not just a problem in Adelaide Hills. PMID- 15831931 TI - Biologics in inflammatory disease: infliximab associated risk of lymphoma development. PMID- 15831932 TI - Hepatocellular carcinoma occurring after successful treatment of childhood cancer with high dose chemotherapy and radiation. PMID- 15831933 TI - Genotypes 677TT and 677CT+1298AC of methylenetetrahydrofolate reductase are associated with the severity of ulcerative colitis in central China. PMID- 15831934 TI - Recognition of mRNA cap structures by viral and cellular proteins. AB - Most cellular and eukaryotic viral mRNAs have a cap structure at their 5' end that is critical for efficient translation. Cap structures also aid in mRNA transport from nucleus to cytoplasm and, in addition, protect the mRNAs from degradation by 5' exonucleases. Cap function is mediated by cap-binding proteins that play a key role in translational control. Recent structural studies on the cellular cap-binding complex, the eukaryotic translation initiation factor 4E and the vaccinia virus protein 39, suggest that these three evolutionary unrelated cap-binding proteins have evolved a common cap-binding pocket by convergent evolution. In this pocket the positively charged N(7)-methylated guanine ring of the cap structure is stacked between two aromatic amino acids. In this review, the similarities and differences in cap binding by these three different cap binding proteins are discussed. A comparison with new functional data for another viral cap-binding protein--the polymerase basic protein (PB2) of influenza virus- suggests that a similar cap-binding mechanism has also evolved in influenza virus. PMID- 15831935 TI - Higher resistance of porcine trigeminal ganglion neurons towards pseudorabies virus-induced cell death compared with other porcine cell types in vitro. AB - Trigeminal ganglion (TG) neurons are important target cells for many alphaherpesviruses, constituting major sites for latency/reactivation events. Here, the in vitro kinetics of productive infection of the swine alphaherpesvirus pseudorabies virus (PRV) and resulting cell death in primary porcine TG neurons were determined, and these were compared with similar kinetics in many other porcine cell types. Confocal microscopy showed that all TG neurons expressed late genes such as viral glycoproteins, and that these glycoproteins were processed through the Golgi and reached the cell surface as in other cell types, albeit with a delay of +/-2-6 h. However, TG neurons were much more resistant towards PRV-induced cell death compared with all other porcine cell types tested (non neuronal TG cells, superior cervical ganglion neurons, epithelial kidney cells, arterial endothelial cells, dermal fibroblasts and cells derived from a porcine swine kidney cell line). About half of the TG neurons survived up to 96 h post inoculation (end of experiment), whereas all other cell types almost completely succumbed within 2 days post-inoculation. In addition, infection with a strongly pro-apoptotic PRV strain that misses the anti-apoptotic US3 protein did not lead to substantial apoptosis in TG neurons, even at 72 h post-inoculation. Thus, primary porcine TG neurons can be infected with PRV in vitro, and are remarkably more resistant to PRV-induced cell death compared with other porcine cell types, suggesting a cell type-specific resistance to alphaherpesvirus-induced cell death that may have important implications for different aspects of the virus life cycle, including latency/reactivation events. PMID- 15831936 TI - Functional interaction of Oct transcription factors with the family of repeats in Epstein-Barr virus oriP. AB - The family of repeats (FR) is a major upstream enhancer of the Epstein-Barr virus (EBV) latent C promoter (Cp) that controls transcription of six different latent nuclear proteins following interaction with the EBV nuclear protein EBNA1. Here, it was shown that Cp could also be activated by octamer-binding factor (Oct) proteins. Physical binding to the FR by the cellular transcription factors Oct-1 and Oct-2 was demonstrated by using an electrophoretic mobility-shift assay. Furthermore, Oct-1 in combination with co-regulator Bob.1, or Oct-2 alone, could drive transcription of a heterologous thymidine kinase promoter linked to the FR in both B cells and epithelial cells. Cp controlled by the FR was also activated by binding of Oct-2 to the FR. This may have direct implications for B cell specific regulation of Cp. PMID- 15831937 TI - Epstein-Barr virus EBNA3 proteins bind to the C8/alpha7 subunit of the 20S proteasome and are degraded by 20S proteasomes in vitro, but are very stable in latently infected B cells. AB - A yeast two-hybrid screen using EBNA3C as bait revealed an interaction between this Epstein-Barr virus (EBV)-encoded nuclear protein and the C8 (alpha7) subunit of the human 20S proteasome. The interaction was confirmed by glutathione S transferase (GST) pull-down experiments and these also revealed that the related proteins EBNA3A and EBNA3B can bind similarly to C8/alpha7. The interaction between these viral proteins and GST-C8/alpha7 was shown to be significantly more robust than the previously reported interaction between C8/alpha7 and the cyclin dependent kinase inhibitor p21(WAF1/CIP1). Co-immunoprecipitation of the EBNA3 proteins with C8/alpha7 was also demonstrated after transfection of expression vectors into B cells. Consistent with this ability to bind directly to an alpha subunit of the 20S proteasome, EBNAs 3A, 3B and 3C were all degraded in vitro by purified 20S proteasomes. However, surprisingly, no sign of proteasome-mediated turnover of these latent viral proteins in EBV-immortalized B cells could be detected, even in the presence of gamma interferon. In actively proliferating lymphoblastoid cell lines, EBNAs 3A, 3B and 3C appear to be remarkably stable, with no evidence of either de novo synthesis or proteasome-mediated degradation. PMID- 15831938 TI - Entry of the vaccinia virus intracellular mature virion and its interactions with glycosaminoglycans. AB - Vaccinia virus (VACV) produces two distinct enveloped virions, the intracellular mature virus (IMV) and the extracellular enveloped virus (EEV), but the entry mechanism of neither virion is understood. Here, the binding and entry of IMV particles have been investigated. The cell receptors for IMV are unknown, but it was proposed that IMV can bind to glycosaminoglycans (GAGs) on the cell surface and three IMV surface proteins have been implicated in this. In this study, the effect of soluble GAGs on IMV infectivity was reinvestigated and it was demonstrated that GAGs affected IMV infectivity partially in some cells, but not at all in others. Therefore, binding of IMV to GAGs is cell type-specific and not essential for IMV entry. By using electron microscopy, it is demonstrated that IMV from strains Western Reserve and modified virus Ankara enter cells by fusion with the plasma membrane. After an IMV particle bound to the cell, the IMV membrane fused with the plasma membrane and released the virus core into the cytoplasm. IMV surface antigen became incorporated into the plasma membrane and was not left outside the cell, as claimed in previous studies. Continuity between the IMV membrane and the plasma membrane was confirmed by tilt-series analysis to orientate membranes perpendicularly to the beam of the electron microscope. This analysis shows unequivocally that IMV is surrounded by a single lipid membrane and enters by fusion at the cell surface. PMID- 15831939 TI - Expression of membrane type 1 matrix metalloproteinase in papillomavirus-positive cells: role of the human papillomavirus (HPV) 16 and HPV8 E7 gene products. AB - Matrix metalloproteinases (MMPs) degrade extracellular matrix. They are involved in cellular proliferation, migration, angiogenesis, invasion and metastasis. MT-1 MMP, a membrane-bound MMP, is expressed in carcinomas of the uterine cervix in vivo. This type of cancer is associated with human papillomavirus (HPV) infection. Here it was shown that keratinocytes transformed with HPV16 or HPV18 in vitro, and HPV-positive cervical carcinoma cell lines, constitutively expressed MT-1 MMP. Expression of the E7 protein from the mucosal and cutaneous high-risk types HPV16 and HPV8, but not from the cutaneous low-risk type HPV1, was sufficient to induce MT-1 MMP expression in primary human keratinocytes and HaCaT cells. As a consequence, MMP-2 was activated. MT-1 MMP expression might play a role in the HPV life cycle by promoting proliferation of host cells and might contribute to their invasive phenotype during malignant progression. PMID- 15831940 TI - Human papillomavirus 16 virus-like particles use heparan sulfates to bind dendritic cells and colocalize with langerin in Langerhans cells. AB - Langerhans cells (LC), the immature dendritic cells (DC) that reside in epithelial tissues are among the first immune cells to encounter human papillomavirus (HPV) and are not activated by HPV virus-like particles (VLPs) in contrast to DC. The notion that the differences in response to HPV VLPs between LC and DC are associated with different types of cell binding and intracellular trafficking has been addressed. Inhibition experiments with heparin and sodium chlorate showed that heparan sulfates are necessary for HPV 16 VLPs to bind to DC but not to LC. Electron microscopy analysis demonstrated a colocalization of HPV 16 VLPs and langerin, which is expressed only by LC. This colocalization was observed on the cell surface but also in cytoplasmic vesicles. As anti-langerin antibodies, HPV 16 VLPs were associated with a faster entry kinetics in LC, as reflected by the fact that VLPs were observed near the nuclear membrane of LC within 10 min whereas more than 60 min were needed in DC. However, no difference between LC and DC was observed for the endocytosis pathway. HPV 16 VLPs entered in both DC and LC by a clathrin-dependent-pathway and were then localized in large cytoplasmic vesicles resembling endosomes. PMID- 15831941 TI - Overlapping reading frames in closely related human papillomaviruses result in modular rates of selection within E2. AB - A core group of four open reading frames (ORFs) is present in all known papillomaviruses (PVs): the E1 and E2 replication/transcription proteins and the L1 and L2 structural proteins. Because they are involved in processes that are essential to PV propagation, the sequences of these proteins are well-conserved. However, sequencing of novel subtypes for human papillomaviruses (HPV) 54 (AE9) and 82 (AE2/IS39), coupled to analysis of four other closely related genital HPV pairs, indicated that E2 has a higher dN/dS ratio than E1, L1 or L2. The elevated ratio is not homogeneous across the length of the ORF, but instead varies with respect to E2's three domains. The E2 hinge region is of particular interest, because its hypervariability (dN/dS>1) differs markedly from the two domains that it joins: the transcription-activation domain and the DNA-binding domain. Deciphering whether the hinge region's high rate of non-synonymous change is the result of positive Darwinian selection or relaxed constraint depends on the evolutionary behaviour of E4, an ORF that overlaps E2. The E2 hinge region is contained within E4 and non-synonymous changes in the hinge are associated with a disproportionate amount of synonymous change in E4, a case of simultaneous positive and purifying selection in overlapping reading frames. Modular rates of selection among E2 domains are a likely consequence of the presence of an embedded E4. E4 appears to be positioned in a part of the HPV genome that can tolerate non-synonymous change and purifying selection of E4 may be indicative of its functional importance. PMID- 15831942 TI - Utility of JC polyomavirus in tracing the pattern of human migrations dating to prehistoric times. AB - JC virus (JCV) is a double-stranded DNA polyomavirus co-evolving with humans since the time of their origin in Africa. JCV seems to provide new insights into the history of human populations, as it suggests an expansion of humans from Africa via two distinct migrations, each carrying a different lineage of the virus. A possible alternative to this interpretation could be that the divergence between the two lineages is due to selective pressures favouring adaptation of JCV to different climates, thus making any inference about human history debatable. In the present study, the evolution of JCV was investigated by applying correspondence analysis to a set of 273 fully sequenced strains. The first and more important axis of ordination led to the detection of 61 nt positions as the main determinants of the divergence between the two virus lineages. One lineage includes strains of types 1 and 4, the other strains of types 2, 3, 7 and 8. The distinctiveness of the Caucasian lineage (types 1 and 4), largely diffused in the northern areas of the world, was almost entirely ascribed to synonymous substitutions. The findings provided by the subsequent axes of ordination supported the view of an evolutionary history of JCV characterized by genetic drift and migration, rather than by natural selection. Correspondence analysis was also applied to a set of 156 human mitochondrial genome sequences. A detailed comparison between the substitution patterns in JCV and mitochondria brought to light some relevant advantages of the use of the virus in tracing human migrations. PMID- 15831943 TI - Role of viral load in the pathogenesis of chicken anemia virus. AB - The pathogenesis of strain 3711 of the chicken anemia virus (CAV), propagated in chickens, and two preparations of strain 3711 that had been adapted to grow to high titre in cells of the MDCC-MSB1 line were studied in chicken embryos and/or chickens. Highest viral loads in infected chickens, as measured by a microplate DNA-hybridization assay, were detected in the thymus, clotted blood and pancreas, and the lowest in the duodenum. The CAV DNA copy number in the organs of chicken embryos was significantly lower than in chickens. Route of infection was an important determinant of the course of disease in chickens, with clinical signs appearing earlier in birds infected by the intramuscular than those infected by the oral route; there was a direct relationship between viral load in particular organs and the extent of clinical signs. No reduction in the pathogenicity for chickens was noted for strain 3711 after 65 or 129 passages in the MDCC-MSB1 cell line. PMID- 15831944 TI - Evidence from nature: interspecies spread of heron hepatitis B viruses. AB - Heron hepatitis B viruses (HHBVs) in three subspecies of free-living great blue herons (Ardea herodias) from Florida, USA, were identified and characterized. Eight of 13 samples were positive in all assays used, whereas sera from egrets, which are also members of the family Ardeidae, were negative in the same assays. Comparative phylogenetic analysis of viral DNA sequences from the preS/S region of previously reported and novel HHBV strains isolated from captive grey herons (Germany) and free-ranging great blue herons (USA), respectively, revealed a strong conservation (95 % sequence similarity) with two separate clusters, implying a common ancestor of all strains. Our data demonstrate for the first time that different subspecies of herons are infected by HHBV and that these infections exist in non-captive birds. Phylogenetic analysis and the fact that the different heron species are geographically isolated populations suggest that lateral transmission, virus adaptation and environmental factors all play a role in HHBV spreading and evolution. PMID- 15831945 TI - Rolling-circle amplification of Torque teno virus (TTV) complete genomes from human and swine sera and identification of a novel swine TTV genogroup. AB - Multiply primed rolling-circle amplification is a novel technology that uses bacteriophage phi29 DNA polymerase to amplify circular DNA molecules, without the need for prior knowledge of their sequences. In an attempt to detect Torque teno virus (TTV), rolling-circle amplification was used to amplify DNA extracted from eight human and four pig serum samples. All samples gave high molecular weight (>30 kb) amplification products. By restriction endonuclease digestion, these products generated DNA fragments whose sizes were consistent with those of human TTV (3.8 kb) and swine TTV (Sd-TTV; 2.9 kb) genomes. Two TTV isolates derived from a single AIDS patient, as well as two Sd-TTV isolates derived from a single pig, were characterized by complete nucleotide sequencing. One of the Sd-TTV isolates showed very low (43-45 %) nucleotide sequence similarity to the other Sd TTV isolate and to the prototype isolate Sd-TTV31, and could be considered the prototype of a novel genogroup. PMID- 15831946 TI - Spodoptera frugiperda resistance to oral infection by Autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion-derived virus binding in the midgut. AB - Spodoptera frugiperda larvae are highly resistant to oral infection by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) (LD(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (LD(50), <1 p.f.u.). The inability of AcMNPV occlusion-derived virus (ODV) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. To determine whether inappropriate binding of AcMNPV ODV to S. frugiperda midgut cells contributes to lack of oral infectivity, the binding and fusion properties of AcMNPV ODV were compared with those of the ODV of a new isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) obtained from a field-collected larva (oral LD(50), 12 occlusions). By using a fluorescence-dequenching assay conducted in vivo, it was found that AcMNPV ODV bound to the midgut epithelia of S. frugiperda larvae at approximately 15 % of the level of SfMNPV ODV, but that, once bound, the efficiencies of fusion for the two ODVs were similar: 60 % for AcMNPV and 53 % for SfMNPV. Whilst the difference in binding efficiencies was significant, it could not account entirely for the observed differences in infectivity. Competition experiments, however, revealed that, in S. frugiperda larvae, SfMNPV ODV bound to a midgut cell receptor that was not bound by AcMNPV ODV, indicating that ODV interaction with a specific receptor(s) was necessary for productive infection of midgut columnar epithelial cells. Fusion in the absence of this ligand-receptor interaction did not result in productive infections. PMID- 15831947 TI - A novel envelope protein involved in White spot syndrome virus infection. AB - One open reading frame (designated vp76) from the White spot syndrome virus (WSSV) genome has the motif of a cytokine I receptor and has been identified as a structural protein. In this paper, vp76 was expressed in Escherichia coli and used to prepare a specific antibody to determine the location of the corresponding protein in the intact virion, the nucleocapsids and the envelope of WSSV. Western blotting with the VP76 antiserum confirmed that VP76 was an envelope protein of WSSV. To investigate the function of the VP76, WSSV was neutralized with the VP76-specific antiserum at different concentrations and injected intramuscularly into crayfish. The mortality curves showed that the VP76 antiserum could partially attenuate infection with WSSV, suggesting that VP76 is an envelope protein involved in WSSV infection. PMID- 15831948 TI - Replication and in vivo repair of the hepatitis A virus genome lacking the poly(A) tail. AB - The precise role of the poly(A) tail at the 3' end of the picornavirus RNA genome and the cellular factors that control its homeostasis are unknown. To assess the importance of the poly(A) tail for virus replication, the genome of the slowly replicating hepatitis A virus (HAV) with and without a poly(A) tail was studied after transfection into cells maintained under various conditions. A tailless HAV genome had a shorter half-life than a poly(A)-containing genome and was unable to replicate in quiescent cells. In dividing cells, the tailless RNA gave rise to infectious virus with a restored poly(A) tail of up to 60 residues. Cells arrested at the G(0) and the G(2)/M phase produced lower amounts of infectious HAV than cells in the G(1) phase. These data suggest that the 3' poly(A) tail of HAV can be restored with the help of a cellular and/or viral function that is regulated during the cell cycle. PMID- 15831949 TI - More recent swine vesicular disease virus isolates retain binding to coxsackie adenovirus receptor, but have lost the ability to bind human decay-accelerating factor (CD55). AB - Swine vesicular disease virus (SVDV) evolved from coxsackie B virus serotype 5 (CVB5) in the recent past, crossing the species barrier from humans to pigs. Here, SVDV isolates from early and recent outbreaks have been compared for their capacity to utilize the progenitor virus receptors coxsackie-adenovirus receptor (CAR) and decay-accelerating factor (DAF; CD55). Virus titre of CVB5 and SVDV isolates It'66 and UK'72 on human HeLa cells was reduced by pre-incubation with either anti-DAF or anti-CAR antibodies; however, recent SVDV isolates R1072, R1120 and SPA'93 did not infect HeLa cells lytically. CVB5 and SVDV infection of the pig cell line IB-RS-2 was inhibited completely by anti-CAR antibodies for all isolates, and no reduction was observed following pre-incubation of cells with anti-pig DAF antibodies. Expression of human DAF in the pig cell line IB-RS-2 enhanced the virus titre of early SVDV isolates by 25-fold, but had no effect on recent SVDV isolate titre. Binding of radiolabelled CVB5 to IB-RS-2 cells was increased seven- to eightfold by expression of human DAF and binding of early SVDV isolates was increased 1.2-1.3-fold, whereas no increase in binding by recent SVDV isolates was mediated by human DAF expression. Addition of soluble hDAF-Fc inhibited CVB5, but not SVDV, infection of pig cells. Pre-incubation of all viruses with soluble hCAR-Fc blocked infection of IB-RS-2 pig cells completely; titration of the amount of soluble hCAR-Fc required to block infection revealed that early isolate UK'72 was the least susceptible to inhibition, and the most recent isolate, SPA'93, was the most susceptible. PMID- 15831950 TI - Apoptotic events induced by human rhinovirus infection. AB - HeLa and 16HBE14o(-) bronchial epithelium cells infected with human rhinovirus serotype 14 (HRV14) were found to exhibit typical apoptotic morphological alterations, such as cell contraction and nuclear condensation. These events coincided with high-molecular-weight DNA fragmentation, activation of caspase-9 and caspase-3 and poly(ADP-ribose) polymerase cleavage. Caspase activation was preceded by cytochrome c translocation from the mitochondria to the cytoplasm, indicating that apoptosis caused by HRV14 infection was triggered predominantly via the mitochondrial pathway. Apoptosis did not affect HRV14 replication per se, but it facilitated the release of newly formed virus from cells. As apoptosis was fully induced at the time of maximal accumulation of progeny HRV14, it is postulated that apoptosis contributed to the destabilization of the cell and facilitated viral progeny release. PMID- 15831951 TI - Temperature-sensitive mutants of enterovirus 71 show attenuation in cynomolgus monkeys. AB - Enterovirus 71 (EV71) is one of the major causative agents of hand, foot and mouth disease and is sometimes associated with serious neurological disorders. In this study, an attempt was made to identify molecular determinants of EV71 attenuation of neurovirulence in a monkey infection model. An infectious cDNA clone of the virulent strain of EV71 prototype BrCr was constructed; temperature sensitive (ts) mutations of an attenuated strain of EV71 or of poliovirus (PV) Sabin vaccine strains were then introduced into the infectious clone. In vitro and in vivo phenotypes of the parental and mutant viruses were analysed in cultured cells and in cynomolgus monkeys, respectively. Mutations in 3D polymerase (3D(pol)) and in the 3' non-translated region (NTR), corresponding to ts determinants of Sabin 1, conferred distinct temperature sensitivity to EV71. An EV71 mutant [EV71(S1-3')] carrying mutations in the 5' NTR, 3D(pol) and in the 3' NTR showed attenuated neurovirulence, resulting in limited spread of virus in the central nervous system of monkeys. These results indicate that EV71 and PV1 share common genetic determinants of neurovirulence in monkeys, despite the distinct properties in their original pathogenesis. PMID- 15831952 TI - Yellow fever virus NS2B-NS3 protease: characterization of charged-to-alanine mutant and revertant viruses and analysis of polyprotein-cleavage activities. AB - A series of 46 charged-to-alanine mutations in the yellow fever virus NS2B-NS3 protease, previously characterized in cell-free and transient cellular expression systems, was tested for their effects on virus recovery. Four distinct plaque phenotypes were observed in cell culture: parental plaque-size (13 mutants), reduced plaque-size (17 mutants), small plaque-size (8 mutants) and no plaque formation (8 mutants). No mutants displayed any temperature sensitivity based on recovery of virus after RNA transfection at 32 versus 37 degrees C. Most small plaque-mutants were defective in growth efficiency compared with parental virus. However not all small plaque-mutants had defective 2B/3 cleavage, with some showing selective defects at other non-structural protein cleavage sites. Revertant viruses were recovered for six mutations that caused reduced plaque sizes. Same-site and second-site mutations occurred in NS2B, and one second-site mutation occurred in the NS3 protease domain. Some reversion mutations ameliorated defects in cleavage activity and plaque size caused by the original mutation. These data indicate that certain mutations that reduce NS2B-NS3 protease cleavage activity cause growth restriction of yellow fever virus in cell culture. However, for at least two mutations, processing defects other than impaired cleavage activity at the 2B/3 site may account for the mutant phenotype. The existence of reversion mutations primarily in NS2B rather than NS3, suggests that the protease domain is less tolerant of structural perturbation compared with the NS2B protein. PMID- 15831953 TI - Mobility of the hepatitis C virus NS4B protein on the endoplasmic reticulum membrane and membrane-associated foci. AB - The hepatitis C virus (HCV) non-structural protein NS4B induces morphological changes in the endoplasmic reticulum (ER) membrane that may have a direct role in viral RNA replication. A chimeric GFP-NS4B fusion protein located to the ER membrane and to foci that were attached to the ER. These membrane-associated foci (MAFs) could be related to the membrane alterations observed in cells that replicate HCV RNA. The relationship of MAFs to pre-existing cellular structures is not known. Indirect immunofluorescence analysis demonstrated that they did not contain a cellular marker for vesicles, which have been implicated in the replication of other viruses. From photobleaching studies to examine diffusion of NS4B, the GFP-tagged protein had reduced mobility on MAFs compared with on the ER membrane. This slower mobility suggested that NS4B is likely to form different interactions on MAFs and the ER. PMID- 15831954 TI - Differential maturation and subcellular localization of severe acute respiratory syndrome coronavirus surface proteins S, M and E. AB - Post-translational modifications and correct subcellular localization of viral structural proteins are prerequisites for assembly and budding of enveloped viruses. Coronaviruses, like the severe acute respiratory syndrome-associated virus (SARS-CoV), bud from the endoplasmic reticulum-Golgi intermediate compartment. In this study, the subcellular distribution and maturation of SARS CoV surface proteins S, M and E were analysed by using C-terminally tagged proteins. As early as 30 min post-entry into the endoplasmic reticulum, high mannosylated S assembles into trimers prior to acquisition of complex N-glycans in the Golgi. Like S, M acquires high-mannose N-glycans that are subsequently modified into complex N-glycans in the Golgi. The N-glycosylation profile and the absence of O-glycosylation on M protein relate SARS-CoV to the previously described group 1 and 3 coronaviruses. Immunofluorescence analysis shows that S is detected in several compartments along the secretory pathway from the endoplasmic reticulum to the plasma membrane while M predominantly localizes in the Golgi, where it accumulates, and in trafficking vesicles. The E protein is not glycosylated. Pulse-chase labelling and confocal microscopy in the presence of protein translation inhibitor cycloheximide revealed that the E protein has a short half-life of 30 min. E protein is found in bright perinuclear patches colocalizing with endoplasmic reticulum markers. In conclusion, SARS-CoV surface proteins S, M and E show differential subcellular localizations when expressed alone suggesting that additional cellular or viral factors might be required for coordinated trafficking to the virus assembly site in the endoplasmic reticulum Golgi intermediate compartment. PMID- 15831955 TI - A single immunization with a rhabdovirus-based vector expressing severe acute respiratory syndrome coronavirus (SARS-CoV) S protein results in the production of high levels of SARS-CoV-neutralizing antibodies. AB - Foreign viral proteins expressed by rabies virus (RV) have been shown to induce potent humoral and cellular immune responses in immunized animals. In addition, highly attenuated and, therefore, very safe RV-based vectors have been constructed. Here, an RV-based vaccine vehicle was utilized as a novel vaccine against severe acute respiratory syndrome coronavirus (SARS-CoV). For this approach, the SARS-CoV nucleocapsid protein (N) or envelope spike protein (S) genes were cloned between the RV glycoprotein G and polymerase L genes. Recombinant vectors expressing SARS-CoV N or S protein were recovered and their immunogenicity was studied in mice. A single inoculation with the RV-based vaccine expressing SARS-CoV S protein induced a strong SARS-CoV-neutralizing antibody response. The ability of the RV-SARS-CoV S vector to confer immunity after a single inoculation makes this live vaccine a promising candidate for eradication of SARS-CoV in animal reservoirs, thereby reducing the risk of transmitting the infection to humans. PMID- 15831956 TI - Analysis of porcine reproductive and respiratory syndrome virus attachment and internalization: distinctive roles for heparan sulphate and sialoadhesin. AB - Heparan sulphate and sialoadhesin were previously identified on porcine macrophages as receptors for porcine reproductive and respiratory syndrome virus (PRRSV). In this study, the exact role and cooperation of heparan sulphate and sialoadhesin during PRRSV attachment and internalization was analysed. It was observed that both heparan sulphate and sialoadhesin mediate PRRSV attachment and that only these two receptors are involved in attachment. Analysis of attachment kinetics of PRRSV to macrophages revealed that early attachment is mediated mainly via an interaction with heparan sulphate, followed by a gradual increase in interaction with sialoadhesin. By using wild-type CHO and CHO deficient in heparan sulphate expression, it was shown that heparan sulphate alone is sufficient to mediate PRRSV attachment, but not entry, and that heparan sulphate is not necessary for sialoadhesin to function as a PRRSV internalization receptor, but enhances the interaction of the virus with sialoadhesin. PMID- 15831957 TI - Generation of recombinant influenza A virus without M2 ion-channel protein by introduction of a point mutation at the 5' end of the viral intron. AB - The aim of this study was to inhibit influenza virus M2 protein expression by mutating the splicing signal of the M gene. Mutations were introduced into the GU dinucleotide sequence at the 5'-proximal splicing site of the M gene (corresponding to nt 52-53 of M cRNA). Transfected cells expressing mutated M viral ribonucleoproteins failed to generate M2 mRNA. Interestingly, recombinant viruses with mutations at the dinucleotide sequence were viable, albeit attenuated, in cell culture. These recombinants failed to express M2 mRNA and M2 protein. These observations demonstrated that the GU invariant dinucleotide sequence at the 5'-proximal splicing site of M gene is essential for M2 mRNA synthesis. These results also indicated that the M2 ion-channel protein is critical, but not essential, for virus replication in cell culture. This approach may provide a new way of producing attenuated influenza A virus. PMID- 15831958 TI - Use of influenza C virus glycoprotein HEF for generation of vesicular stomatitis virus pseudotypes. AB - Influenza C virus contains two envelope glycoproteins: CM2, a putative ion channel protein; and HEF, a unique multifunctional protein that performs receptor binding, receptor-destroying and fusion activities. Here, it is demonstrated that expression of HEF is sufficient to pseudotype replication-incompetent vesicular stomatitis virus (VSV) that lacks the VSV glycoprotein (G) gene. The pseudotyped virus showed characteristic features of influenza C virus with respect to proteolytic activation, receptor usage and cell tropism. Chimeric glycoproteins composed of HEF ectodomain and VSV-G C-terminal domains were efficiently incorporated into VSV particles and showed receptor-binding and receptor destroying activities but, unlike authentic HEF, did not mediate efficient infection, probably because of impaired fusion activity. HEF-pseudotyped VSV efficiently infected polarized Madin-Darby canine kidney cells via the apical plasma membrane, whereas entry of VSV-G-complemented virus was restricted to the basolateral membrane. These findings suggest that pseudotyping of viral vectors with HEF might be useful for efficient apical gene transfer into polarized epithelial cells and for targeting cells that express 9-O-acetylated sialic acids. PMID- 15831959 TI - Evolutionary timescale of rabies virus adaptation to North American bats inferred from the substitution rate of the nucleoprotein gene. AB - Throughout North America, rabies virus (RV) is endemic in bats. Distinct RV variants exist that are closely associated with infection of individual host species, such that there is little or no sustained spillover infection away from the primary host. Using Bayesian methodology, nucleotide substitution rates were estimated from alignments of partial nucleoprotein (N) gene sequences of nine distinct bat RV variants from North America. Substitution rates ranged from 2.32 x 10(-4) to 1.38 x 10(-3) substitutions per site per year. A maximum-likelihood (ML) molecular clock model was rejected for only two of the nine datasets. In addition, using sequences from bat RV variants across the Americas, the evolutionary rate for the complete N gene was estimated to be 2.32 x 10(-4). This rate was used to scale trees using Bayesian and ML methods, and the time of the most recent common ancestor for current bat RV variant diversity in the Americas was estimated to be 1660 (range 1267-1782) and 1651 (range 1254-1773), respectively. Our reconstructions suggest that RV variants currently associated with infection of bats from Latin America (Desmodus and Tadarida) share the earliest common ancestor with the progenitor RV. In addition, from the ML tree, times were estimated for the emergence of the three major lineages responsible for bat rabies cases in North America. Adaptation to infection of the colonial bat species analysed (Eptesicus fuscus, Myotis spp.) appears to have occurred much quicker than for the solitary species analysed (Lasionycteris noctivagans, Pipistrellus subflavus, Lasiurus borealis, Lasiurus cinereus), suggesting that the process of virus adaptation may be dependent on host biology. PMID- 15831960 TI - Polarized entry and release of Junin virus, a New World arenavirus. AB - Junin virus (JUNV), the causative agent of Argentine haemorrhagic fever, is a human pathogen that naturally enters the body through the epithelial cells of the respiratory and digestive tracts. The interaction of JUNV with two types of polarized epithelial cultures, Vero C1008 and A549, was investigated. Radioactive virus-binding assays showed that JUNV infects polarized lines preferentially through the apical surface. High-level expression of viral nucleoprotein was detected in polarized cell lines infected through the apical domain. Virus production from apical media was about 100-fold higher than that found into the basolateral medium. Confocal-immunofluorescence analysis revealed high-level expression of glycoprotein at the apical-membrane surface. Disruption of the microtubule network by colchicine impaired JUNV vectorial release. This is the first study to analyse the interaction between a member of the virus family Arenaviridae and polarized epithelial cells, showing preferential entry and release from the apical plasma membrane. PMID- 15831961 TI - RNA interference of rotavirus segment 11 mRNA reveals the essential role of NSP5 in the virus replicative cycle. AB - Rotavirus genomes contain 11 double-stranded (ds) RNA segments. Genome segment 11 encodes the non-structural protein NSP5 and, in some strains, also NSP6. NSP5 is produced soon after viral infection and localizes in cytoplasmic viroplasms, where virus replication takes place. RNA interference by small interfering (si) RNAs targeted to genome segment 11 mRNA of two different strains blocked production of NSP5 in a strain-specific manner, with a strong effect on the overall replicative cycle: inhibition of viroplasm formation, decreased production of other structural and non-structural proteins, synthesis of viral genomic dsRNA and production of infectious particles. These effects were shown not to be due to inhibition of NSP6. The results obtained strengthen the importance of secondary transcription/translation in rotavirus replication and demonstrate that NSP5 is essential for the assembly of viroplasms and virus replication. PMID- 15831962 TI - Correlation between interferon sensitivity of reovirus isolates and ability to discriminate between normal and Ras-transformed cells. AB - Mammalian reoviruses exhibit a propensity to replicate in transformed cells. It is currently believed that the interferon-inducible RNA-dependent protein kinase (PKR), an intracellular host-cell resistance factor that is inhibited by an activated Ras-dependent pathway in transformed cells, is responsible for this discrimination. In the present study, reovirus isolates differing in their sensitivity to interferon were obtained by chemical mutagenesis, and examined for their replicative properties in parental and Ras-transformed mouse NIH-3T3 cells. It was observed that most isolates can bypass resistance mechanisms of parental cells at high m.o.i., and that there is a correlation between the ability to discriminate between transformed and parental cells, and interferon sensitivity. Most interestingly, an interferon-hypersensitive mutant virus was more dependent on Ras activation than any other viral isolate. Altogether, this suggests that optimal reovirus isolates could be selected to attack tumour cells depending on the nature of the alterations in interferon-inducible pathways found in these cells. PMID- 15831963 TI - An antibody specific for the C-terminal tail of the gp41 transmembrane protein of human immunodeficiency virus type 1 mediates post-attachment neutralization, probably through inhibition of virus-cell fusion. AB - Evidence has been presented which shows that part of the C-terminal tail of the gp41 transmembrane protein of human immunodeficiency virus type 1 (HIV-1) contains a neutralization epitope and is thus exposed on the external surface of the virion. Here, SAR1, a monoclonal antibody, which was stimulated by immunization with a plant virus expressing 60 copies of the GERDRDR sequence from the exposed gp41 tail, and has an unusual pattern of neutralization activity, giving little or no neutralization of free virions, but effecting modest post attachment neutralization (PAN) of virus bound to target cells was investigated. Here, the properties of PAN were investigated. It was found that PAN could be mediated at 4 or 20 degrees C, but that at 20 degrees C maximum PAN required virus-cell complexes to be incubated for 3 h before addition of antibody. Further PAN appeared stable at 20 degrees C and could be mediated for at least 5 h at this temperature. In contrast, when virus-cell complexes formed at 20 degrees C but then shifted to 37 degrees C for various times before addition of SAR1, PAN was maximal after just 10 min, and was lost after 30 min incubation. Thus, PAN at 37 degrees C is transient and temperature-dependent. Since this scenario recalled the temperature requirements of virus-cell fusion, fusion of HIV-1-infected and non-infected cells was investigated, and it was found that SAR1 inhibited this process by up to 75 %, in a dose-dependent manner. However, antibodies to adjacent epitopes did not inhibit fusion. These data confirm the external location of the SAR1 epitope, implicate the gp41 C-terminal tail in the HIV-1 fusion process for the first time, and suggest that SAR1 mediates PAN by inhibiting virus-mediated fusion. PMID- 15831964 TI - Phenotypic analysis of human immunodeficiency virus type 1 Rev trimerization interface mutants in human cells. AB - Nuclear export of unspliced and incompletely spliced human immunodeficiency virus type 1 mRNA is mediated by the viral Rev protein. Rev binds to a structured RNA motif known as the Rev-response element (RRE), which is present in all Rev dependent transcripts, and thereby promotes entry of the ribonucleoprotein complex into the nuclear-export pathway. Recent evidence indicates that a dimerization interface and a genetically separable "trimerization" interface are required for multimeric assembly of Rev on the RRE. In this report, the effect of mutations within the trimerization interface on Rev function was examined in mammalian cells. All trimerization-defective Rev molecules had profoundly compromised Rev function and a range of localization defects was observed. However, despite the potential for formation of heterodimers between functional and non-functional Rev proteins, trimerization-defective Rev mutants were unable to inhibit wild-type Rev function in a trans-dominant-negative manner. PMID- 15831965 TI - A functional CD8+ cell assay reveals individual variation in CD8+ cell antiviral efficacy and explains differences in human T-lymphotropic virus type 1 proviral load. AB - The CD8+ lymphocyte response is a main component of host immunity, yet it is difficult to quantify its contribution to the control of persistent viruses. Consequently, it remains controversial as to whether CD8+ cells have a biologically significant impact on viral burden and disease progression in infections such as human immunodeficiency virus-1 and human T-lymphotropic virus type I (HTLV-I). Experiments to ascertain the impact of CD8+ cells on viral burden based on CD8+ cell frequency or specificity alone give inconsistent results. Here, an alternative approach was developed that directly quantifies the impact of CD8+ lymphocytes on HTLV-I proviral burden by measuring the rate at which HTLV-I-infected CD4+ cells were cleared by autologous CD8+ cells ex vivo. It was demonstrated that CD8+ cells reduced the lifespan of infected CD4+ cells to 1 day, considerably shorter than the 30 day lifespan of uninfected cells in vivo. Furthermore, it was shown that HTLV-I-infected individuals vary considerably in the rate at which their CD8+ cells clear infected cells, and that this was a significant predictor of their HTLV-I proviral load. Forty to 50 % of between-individual variation in HTLV-I proviral load was explained by variation in the rate at which CD8+ cells cleared infected cells. This novel approach demonstrates that CD8+ cells are a major determinant of HTLV-I proviral load. This assay is applicable to quantifying the CD8+ cell response to other viruses and malignancies and may be of particular importance in assessing vaccines. PMID- 15831966 TI - Differential transcriptional activity of plant-pathogenic begomoviruses in their whitefly vector (Bemisia tabaci, Gennadius: Hemiptera Aleyrodidae). AB - Plant-pathogenic begomoviruses have a complex association with their whitefly vector and aspects concerning virus genetic activity (genome replication and gene transcription) within the insect remain highly controversial. Virus transcript abundance was assessed by quantifying selected gene transcripts of Tomato mottle virus (ToMoV, a New World bipartite begomovirus) and Tomato yellow leaf curl virus (TYLCV, an Old World monopartite begomovirus) in whiteflies (Bemisia tabaci biotype B) after feeding on virus-infected tomato plants and after subsequent transfer to cotton, a plant that is immune to the selected begomoviruses. Real time RT-PCR was performed using specific primers for three ToMoV genes (AV1, BC1 and BV1) and three TYLCV genes (V1, V2 and C3). The ToMoV gene transcripts rapidly became undetectable in whiteflies following transfer from tomato to cotton, probably because degradation was not accompanied by new synthesis. On the other hand, TYLCV transcripts increased after transfer of whiteflies to cotton, indicating active TYLCV transcription. Interestingly, the difference observed in ToMoV and TYLCV transcripts in the vector parallel observations on the different biological effects of these viruses on whiteflies, i.e. TYLCV, but not ToMoV, reduces whitefly fitness. PMID- 15831967 TI - South West Indian Ocean islands tomato begomovirus populations represent a new major monopartite begomovirus group. AB - Biological and molecular properties of Tomato leaf curl Madagascar virus isolates from Morondova and Toliary (ToLCMGV-[Tol], -[Mor]), Tomato leaf curl Mayotte virus isolates from Dembeni and Kahani (ToLCYTV-[Dem], -[Kah]) and a Tomato yellow leaf curl virus isolate from Reunion (TYLCV-Mld[RE]) were determined. Full length DNA components of the five isolates from Madagascar, Mayotte and Reunion were cloned and sequenced and, with the exception of ToLCMGV-[Tol], were shown to be both infectious in tomato and transmissible by Bemisia tabaci. Sequence analysis revealed that these viruses had genome organizations of monopartite begomoviruses and that both ToLCMGV and ToLCYTV belong to the African begomoviruses but represent a distinct monophyletic group that we have tentatively named the South West islands of the Indian Ocean (SWIO). All of the SWIO isolates examined were apparently complex recombinants. None of the sequences within the recombinant regions closely resembled that of any known non SWIO begomovirus, suggesting an isolation of these virus populations. PMID- 15831968 TI - Reciprocal function of movement proteins and complementation of long-distance movement of Cymbidium mosaic virus RNA by Odontoglossum ringspot virus coat protein. AB - Complementation of movement and coat proteins of the orchid-infecting potexvirus Cymbidium mosaic virus (CymMV) and tobamovirus Odontoglossum ringspot virus (ORSV) was investigated. Nicotiana benthamiana, which is susceptible to both CymMV and ORSV, was used as a model system. Four transgenic lines, each harbouring one of the movement protein (MP) or coat protein (CP) genes of CymMV or ORSV, were constructed. The MP of CymMV consists of three overlapping open reading frames, together called the triple-gene block (TGB). CymMV and ORSV mutants, each carrying an inactivated MP or CP, were generated from the respective biologically active full-length cDNA clones. Complementation was studied by infecting transgenic plants with in vitro transcripts generated from these mutants. The cell-to-cell movement of a movement-deficient CymMV was restored in transgenic plants carrying the ORSV MP transgene. Similarly, CymMV TGB1 transgenic plants were able to rescue the cell-to-cell movement of a movement-deficient ORSV mutant. ORSV CP transgenic plants supported systemic movement of a CymMV CP-deficient mutant. However, in these plants, neither encapsidation of CymMV RNA with ORSV CP nor CymMV CP expression was detected. Long-distance movement of an ORSV CP-deficient mutant was not supported by CymMV CP. The complementation of MPs and CPs of CymMV and ORSV facilitates movement of these viruses in plants, except for long-distance movement of ORSV RNA by CymMV CP. PMID- 15831969 TI - Complete genome sequences of two new variants of Grapevine rupestris stem pitting associated virus and comparative analyses. AB - Grapevine rupestris stem pitting-associated virus (GRSPaV), a member of the genus Foveavirus within the family Flexiviridae, is the putative causal agent of the disease Rupestris stem pitting (RSP) of grapevines. GRSPaV comprises a family of variants whose pathological characteristics have not been determined. Recently, many of the indicator "St George" plants (Vitis rupestris) used throughout the world to index RSP tested positive for GRSPaV. This finding questions the validity of past biological indexing results. In this work, a representative genomic region of GRSPaV was first sequenced from ten "St George" plants from two sources and it was demonstrated that nine of them carried a new variant, GRSPaV SG1. The genomes of GRSPaV-SG1 and GRSPaV-BS from "Bertille Seyve 5563" plants were sequenced, revealing a genome structure identical to that of GRSPaV-1. It was demonstrated experimentally that infection of "St George" plants with GRSPaV SG1 is asymptomatic and thus it is proposed that GRSPaV-SG1 infection should not have interfered with the outcome of past indicator indexing. This represents the first attempt to link a GRSPaV variant with pathological properties. PMID- 15831970 TI - A double-stranded RNA from a Phytophthora species is related to the plant endornaviruses and contains a putative UDP glycosyltransferase gene. AB - A new dsRNA was isolated from a Phytophthora isolate from Douglas fir. Sequence analysis showed the dsRNA to consist of 13 883 bp and to contain a single open reading frame with the potential to encode a polyprotein of 4548 aa. This polyprotein contained amino acid sequence motifs characteristic of virus RNA dependent RNA polymerases (RdRps) in its C-terminal region and motifs characteristic of RNA helicases in its N-terminal region. These sequence motifs were related to corresponding motifs in plant viruses in the genus Endornavirus. In phylogenetic trees constructed from the RdRp and helicase motifs of a range of ssRNA and dsRNA viruses, the Phytophthora RdRp and helicase sequences clustered with those of the plant endornaviruses with good bootstrap support. The properties of the Phytophthora dsRNA are consistent with its being classified as the first non-plant member of the genus Endornavirus, for which we propose the name phytophthora endornavirus 1 (PEV1). A region between the RdRp and helicase domains of the PEV1 protein had significant amino acid sequence similarity to UDP glycosyltransferases (UGTs). Two sequence motifs were identified, one characteristic of all UGTs and the other characteristic of sterol UGTs. The PEV1 UGT would be the first for an RNA virus, although ecdysteroid UGT genes have been found in many baculoviruses. The PEV1 UGT was only distantly related to baculovirus ecdysteroid UGTs, which belong to a family distinct from the sterol UGTs. PMID- 15831971 TI - Expression of PrPC on cellular components of sheep blood. AB - PrP(C), a glycosylphosphatidylinositol-linked glycoprotein, plays a central role in the pathogenesis of transmissible spongiform encephalopathies (TSEs), undergoing a conformational alteration to the disease-associated isoform, commonly designated PrP(Sc). PrP(C) is expressed in many tissues other than the nervous system, although its precise function(s) remains unclear. It has previously been demonstrated that TSEs can be transmitted by blood transfusion in sheep. The aim of this work was to identify which components of blood carried the infection. As an initial step, the distribution of PrP(C) on cellular components of sheep blood was examined to identify potential targets for infection. Cell surface expression of PrP(C) was found only on peripheral blood mononuclear cells (PBMCs); however, platelets also contained significant amounts of intracellular PrP(C). The level of PrP(C) expressed on the cell surface of PBMCs was influenced by PrP genotype, with the highest levels found in scrapie-susceptible VRQ/VRQ sheep and the lowest levels in scrapie-resistant ARR/ARR sheep. In susceptible sheep, PrP(C) was expressed at varying levels on all major subsets of PBMCs, with the highest levels on the CD21(+) subset of B cells, and PrP expression was upregulated dramatically on CD21(+) B cells in some scrapie-infected sheep. PMID- 15831972 TI - Evaluation of the Coroflex Theca-Stent for reduction of restenosis (ECORI). AB - BACKGROUND: Various stent coatings have been shown to significantly reduce restenosis rates in comparison to non-coated devices. Therefore, the short- and mid-term performance of the new polyphosphazene-coated Coroflex Theca-Stent was investigated. METHODS: 103 patients [63.9 +/- 11 yrs, 5/103 (4.9%) lesion type A, 52/103 (50.5%) type B1, and 46 of 103 (44.6%) type B2] were enrolled for elective single stent deployment into de-novo coronary lesions (stenoses: greater than or equal to 70%, < 100%; reference diameter greater than or equal to 2.75 mm, less than or equal to 4 mm; lesion length: < 16 mm). RESULTS: Deployment and procedural success were 100%, in 57/103 (55.3%) patients without pre-dilatation. 3/103 (2.9%) patients were lost to follow-up. During the 7.1 +/- 2.3 months clinical follow-up, 3 of 100 (3.0%) patients underwent premature target lesion revascularizations, 4 /100 (4%) had non-target lesion-related deaths, and 1 of 100 (1%) suffered myocardial infarction. Among the 77 of 100 (77.0%) patients who underwent angiographic follow-up, the initial stenosis declined from 87.3 +/- 5.7% to 14.2 +/- 8.3% after stenting, and increased to 32.8 +/- 22.7% after 6.4 +/- 1.3 months. The late loss and late loss index were 0.6 +/- 0.7 mm and 0.2 +/- 0.4, respectively; the recurrence rate was 12 of 77 (15.6%), with reintervention required in 11 of 77 (14.3%) of these patients. CONCLUSION: The Coroflex Theca Stent provides excellent procedural results and a low restenosis rate. Further development of this polymer as the final coating and as the basis for drug eluting stents seems justified. PMID- 15831973 TI - Effects of focused force angioplasty: pre-clinical experience and clinical confirmation. AB - Balloon angioplasty results in a combination of plaque compression and fracture, creation of intimal flaps and localized medial dissection, as well as the stretching and remodeling of the arterial wall. The application of eccentric pressure to the vessel wall allows for the balloon force to be focused along a specified line resulting in a reduction in vessel resistance with an improved or equivalent result at a lower pressure. The FX miniRAIL is a novel balloon angioplasty catheter that has two external wires that are compressed by the inflating balloon into the vessel wall. It was tested in comparison to conventional balloon injury in a porcine coronary model. At equivalent balloon artery ratios no increase in the length of dissections occurred, but a characteristic longitudinal "cut" was noted in the presence of the FX miniRAIL. A similar finding was demonstrated in sample human cases when interrogated by IVUS and OCT. PMID- 15831974 TI - Expanded applications of rotational atherectomy in contemporary coronary and peripheral interventional practice. AB - Percutaneous rotational atherectomy is mainly utilized in contemporary interventional practice to alter lesion compliance, facilitating stent delivery and antirestenotic drug delivery at the site of the underlying lesion. This enables a percutaneous revascularization strategy in a group of patients who would otherwise require a surgical revascularization. We identify and present three novel uses for this device in percutaneous coronary and peripheral interventional procedures, which further expands the applications of rotational atherectomy. PMID- 15831975 TI - Iso-osmolar radio contrast iodixanol in patients with chronic kidney disease. AB - BACKGROUND: Although radio contrast volume has been associated with worsening post-procedural kidney function, this relationship has not been extensively studied using an iso-osmolar contrast agent in chronic kidney disease patients. METHODS: We retrospectively studied patients undergoing cardiac catheterization at the University of Minnesota from 2000 to 2004, using the iso-osmolar contrast agent, iodixanol. All patients were included who had calculated creatinine clearance (CCR) < 60 mL/min, not on dialysis, and serum creatinine measured on the same day and within 7 days after the procedure. Comparison of a subgroup with severe chronic kidney disease and diabetes mellitus was compared to a similar historical control group that used the low-osmolar contrast agent, iohexol. RESULTS: Serum creatinine and CCR were 2.9 +/- 1.5 mg/dL and 33.4 +/- 12.0 mL/min (mean +/- standard deviation), respectively, at baseline in 117 cases. Peak creatinine increased by 0.03 +/- 0.7 mg/dL after 84.3 +/- 67.3 mL of iodixanol was used. Contrast-induced nephropathy definition was fulfilled in 22 (18.8%) cases. A non-significant negative correlation was found between the volume of iodixanol and the change in creatinine (r2 = 0.0011, p = 0.7254). A subgroup with severe chronic kidney disease and diabetes mellitus with iodixanol had a significantly lower creatinine increase (n = 25, 0.09 +/- 0.5 mg/dL), compared to historical controls (n = 42, 0.7 +/- 0.8 mg/dL) with iohexol (p < 0.001). A non significant positive correlation between volume of contrast and change in creatinine was found in this subgroup who received iodixanol (n = 25, r2 = 0.0756, p = 0.1835), but was significant in the historical controls who received iohexol (n = 42, r2 = 0.135, p = 0.017). CONCLUSIONS: The volume of iso-osmolar radio contrast does not affect the incidence of contrast-induced nephropathy in patients with chronic kidney disease. A randomized trial evaluating the incidence of contrast nephropathy would verify the safety of ad hoc versus staged angiographic procedures in this population. PMID- 15831976 TI - Contrast-induced nephropathy and cardiac catheterization: evidence in support of using the iso-osmolar contrast agent iodixanol in patients with renal dysfunction. PMID- 15831977 TI - Long-term outcome of patients with proximal left anterior descending coronary artery in-stent restenosis treated with a second percutaneous procedure. AB - Patients with in-stent restenosis involving the proximal segment of the left anterior descending coronary artery are frequently sent for surgical revascularization. We studied the long-term (3.3 +/- 2.8 years) outcome of 123 patients with in-stent restenosis involving the proximal left anterior descending coronary artery who were treated with a second percutaneous procedure. Cumulative survival rate, probability of being alive and free from surgical revascularization, and probability of being alive and free from new revascularization procedures was 98.3 +/- 1.2%, 92.8 +/- 2.5%, and 82.2 +/- 3.6% at 1 year, and 95.2 +/- 2.5%, 88.5 +/- 3.4%, and 76.9 +/- 4.3% at 3 years, respectively. Thus, in patients with in-stent restenosis involving the proximal left anterior descending artery, a second percutaneous procedure is a feasible and safe long-term strategy, with few patients ultimately requiring surgical revascularization. PMID- 15831978 TI - Passive stent coatings in the drug-eluting era. AB - Advances in coronary stent technology, both in terms of stent design and function, have significantly improved the safety and efficacy of percutaneous coronary intervention, including marked reduction in restenosis. This has led to its use in increasingly challenging clinical and lesional subsets, with potential for increased risk of stent-associated thrombosis. In this article we outline the development of passive stent coatings, and evaluate the ongoing role of such coatings in the contemporary era of antiproliferative drug-eluting stents. PMID- 15831979 TI - Post-stent very proximal left anterior descending coronary artery aneurysm. PMID- 15831980 TI - Catheter-induced left main dissection incidence, predisposition and therapeutic strategies experience from two sides of the hemisphere. PMID- 15831981 TI - The International Andreas Gruentzig Society: 20 years of progress on cardiac and vascular intervention--a legacy to Andreas. PMID- 15831982 TI - Vancomycin-resistant enterococcal bacteremia in a hematology unit: molecular epidemiology and analysis of clinical course. AB - An increase in vancomycin-resistant enterococcal (VRE) bacteremia in hemato oncological patients (n=19) in our institution from 2000 through 2001 led us to analyze the molecular epidemiologic patterns and clinical features unique to our cases. The pulsed field gel electrophoresis of the isolates revealed that the bacteremia was not originated from a single clone but rather showed endemic pattern of diverse clones with small clusters. A different DNA pattern of blood and stool isolates from one patient suggested exogenous rather than endogenous route of infection. Enterococcus faecium carrying vanA gene was the causative pathogen in all cases. Patients with VRE bacteremia showed similar clinical courses compared with those with vancomycin-susceptible enterococcal (VSE) bacteremia. Vancomycin resistance did not seem to be a poor prognostic factor because of similar mortality (5/8, 62.5%) noted in VSE bacteremia. Initial disease severity and neutropenic status may be major determinants of prognosis in patients with VRE bacteraemia. PMID- 15831983 TI - The efficacy of clinical strategies to reduce nosocomial sepsis in extremely low birth weight infants. AB - The aim of this study was to evaluate the efficacy of clinical strategies to reduce nosocomial sepsis (NS) in extremely low birth weight infants (ELBWI) less than 1,000 g. Data from the period before (P1, 1995-2000) and after (P2, 2001 2002) implementation of the strategies were collected and analyzed. The intervention strategies included restriction of antibiotic therapy, less use of invasive procedures such as umbilical vessel catheterization and endotracheal intubation, establishment of guidelines for hand-washing, infant handling, and central intravascular line management. NS was defined as positive blood cultures in symptomatic patients after the third day of life with the use of antibiotics for more than 5 days. Although the gestational age (GA) and birth weight (BW) were significantly lower in P2 (GA 26.7 +/-2.1 wk; BW 796 +/-130 g) compared to P1 (GA 27.2 +/-1.6 wk; BW 857 +/-121 g), the incidence of NS decreased significantly from 70% (69/99) in P1 to 17% (24/71) in P2 with the implementation of the intervention strategies. The coagulase negative Staphylococcus infection was also significantly reduced from 34% in P1 to 11% in P2. The implementation of the clinical strategies was quite effective in reducing the incidence of NS in ELBWI. PMID- 15831984 TI - Low birth weight,very low birth weight rates and gestational age-specific birth weight distribution of korean newborn infants. AB - To obtain the low birth weight (LBW) rate, the very low birth weight (VLBW) rate, and gestational age (GA)-specific birth weight distribution based on a large population in Korea, we collected and analyzed the birth data of 108,486 live births with GA greater than 23 weeks for 1 yr from 1 January to 31 December 2001, from 75 hospitals and clinics located in Korea. These data included birth weight, GA, gender of the infants, delivery type, maternal age, and the presence of multiple pregnancy. The mean birth weight and GA of a crude population are 3,188 +/-518 g and 38.7+/-2.1 weeks, respectively. The LBW and the VLBW rates are 7.2% and 1.4%, respectively. The preterm birth rate (less than 37 completed weeks of gestation) is 8.4% and the very preterm birth rate (less than 32 completed weeks of gestation) is 0.7%. The mean birth weights for female infants, multiple births, and births delivered by cesarean section were lower than those for male, singletons, and births delivered vaginally. The risk of delivering LBW or VLBW infant was higher for the teenagers and the older women (aged 35 yr and more). We have also obtained the percentile distribution of GA-specific birth weight in infants over 23 weeks of gestation. PMID- 15831985 TI - A retrospective cohort study on obesity and hypertension risk among Korean adults. AB - A retrospective cohort of adult Korean males and females was conducted to evaluate the influence of obesity on the development of hypertension and to determine the level of the body mass index at which the risk of hypertension significantly increases. The subjects of this study were 1,467 men and 944 women aged 20 to 75 yr who were normotensive at the time of their initial examinations from 1990 to 1991, who had a follow-up examination at least 1 yr after their initial examinations, or whose blood pressure status could be confirmed by reviewing their medical records until June 2000. During an average follow-up period of 6.2 yr, 234 new cases of hypertension were identified. An analysis using the Cox proportional hazards model showed that the risk of developing hypertension increased with increasing age, body mass index, and amount of daily alcohol consumption in men; and with increasing age and body mass index in women. Comparing men and women whose body mass indices were smaller than 23 kg/m(2), the relative risks of hyper-tension were 2.56 times greater in men and 3.17 times greater in women, whose body mass indices were greater than 27 kg/m(2). Our study confirmed that obesity is a strong risk factor for hypertension among Korean adults. In addition, high alcohol consumption may be a significant risk factor for men. PMID- 15831986 TI - Percutaneous coronary intervention versus coronary artery bypass grafting for diabetics with multivessel coronary artery disease: the Korean Multicenter Revascularization Registry (KORR). AB - This study was designed to assess the relative merits of percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) in multivessel coronary artery disease (MVCAD), particularly for Korean diabetics. Among 3,279 patients with MVCAD who were recommended for revascularization were enrolled from nine centers in Korea, 2,154 were selected after statistical adjustments for the disparities between two groups. Survival rates were not significantly different for three years between two groups. Among diabetic patients, the three-year mortality rate in PCI group was 1.9-fold higher than that of CABG group, although it was not statistically significant (PCI 19.8%, CABG 11.4%, p=0.14). The three year mortality rate was similar between the two groups in non-diabetics (PCI 8.3%, CABG 10.0%, p=0.50). The 30-day rate of cerebrovascular event was higher in CABG group, for both diabetic (CABG 3.6%, PCI 0.0%, p<0.001) and non-diabetic patients (CABG 2.4%, PCI 0.0%, p<0.001). Short- and long-term revascularization rates were higher in PCI group than in CABG group. As a conclusion, this Korean registry demonstrates that PCI was associated with comparable survival rates and lower short-term morbidity, but a greater requirement for repeated revascularization compared with CABG in Korean diabetics. PMID- 15831987 TI - Changes of coronary blood flow in vasospastic angina under cold stimulation by transthoracic Doppler echocardiography. AB - This study was done to evaluate changes of microvascular function under cold stimulation by measuring coronary flow velocities (CFVs) in vasospastic angina (VA) patients using transthoracic Doppler echocardiography (TTDE). 14 patients with VA and 15 healthy controls were included. CFVs were measured at the distal left anterior descending coronary artery by TTDE at baseline and under cold stimulation. Hyperemia was induced by intravenous adenosine infusion (140 microg/kg/min). At baseline, CFVs and coronary flow reserve (CFR) were not different between controls and VA patients. Under cold stimulation, the degree of increment of CFV with adenosine was lower in VA patients than in controls. Comparing baseline with cold stimulation, coronary flow reserve (CFR) increased (3.1+/-0.7 to 3.8 +/-1.0, p=0.06) in controls. In contrast, in VA patients, CFR was decreased (2.8 +/-0.9 to 2.6 +/-0.7, p=0.05) and coronary vascular resistance index markedly increased (0.35 to 0.43, p=0.01). Throughout the study, no patient experienced chest pain or ECG changes. In VA patients, CFR was preserved at baseline, but coronary blood flow increase in response to cold stimulation was blunted and CFR was decreased. These findings suggest that endothelial dependent vasodilation is impaired at the coronary microvascular and the epicardial artery level in VA under cold stimulation. PMID- 15831988 TI - Successful treatment of Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis with HLH-94 protocol. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare, fatal disorder of children, affecting predominantly the mononuclear phagocytic system. Previous reports indicate that Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (EBV-HLH) can also be fatal in many cases, although the prognosis for EBV-HLH is better than for the familial form of hemophagocytic lymphohistiocytosis. We treated four patients with EBV-HLH using immunochemotherapy including steroid, etoposide (VP-16), and cyclosporin, according to the HLH-94 protocol. All patients experienced persistent fever, cytopenia, and hypertriglyceridemia. Serological testing for EBV showed reactivated EBV infections in all patients. EBV DNA detected by PCR and EBV encoded small RNA measured by in situ hybridization were confirmed in the patients' bone marrow specimens. Hemophagocytosis was shown in bone marrow aspirates and liver biopsy specimen. Complete remission was achieved in all patients after induction and continuation therapy for 4-10 months (median, 7 months) and was maintained for 15-27 months (median, 19 months) without the need for bone marrow transplantation. These results suggest that EBV-HLH can be effectively controlled by immunochemotherapy using the HLH-94 protocol. PMID- 15831989 TI - Lymphocyte subsets and mood states in panic disorder patients. AB - This study was conducted to examine lymphocyte subset counts and mood states in panic disorder patients. Twenty patients with panic disorder and 20 age- and gender-matched normal healthy subjects were recruited for the study. We used the Spielberger State (STAIS) & Trait (STAIT) Anxiety Inventory, Hamilton Depression Rating scale (HAMD) and Hamilton Anxiety Rating scale (HAMA) to measure mood states in all subjects. Lymphocyte subsets counts were made by flow cytometry. Panic patients showed significantly higher scores for anxiety and depression than normal subjects. Panic patients showed no differences in terms of the numbers of immune cells, as compared with normal healthy subjects, other than a lower proportion of T suppressor cells and a higher T helper cell/T suppressor cell ratio. HAMA and STAIS scores were common factors that could predict T cell numbers and proportions, T helper cell numbers, and natural killer cell proportions in panic disorder patients. We suggest that anxiety levels are related to the T-cell population in panic disorder patients and that quantitative immune differences may reflect altered immunity in this disorder. PMID- 15831990 TI - Two novel mutations in the C7 gene in a Korean patient with complement C7 deficiency. AB - Complement C7 deficiency is an autosomal recessive disorder well known to be associated with increased susceptibility to meningococcal infection and has mostly been reported in Caucasians. In the Korean population, no case of C7 deficiency has been reported to date. Recently we experienced an 11-yr-old girl with meningococcal meningitis who was diagnosed as having C7 deficiency based upon the undetectable serum C7 protein on radial immunodiffusion and the undetectable serum total and C7 hemolytic activities. To identify the genetic basis of the C7 deficiency of the patient, we performed a mutation analysis for the C7 gene and found two novel mutations; a point mutation at the 3' splice acceptor site of intron 4 (c.281-1G>T) and a large deletion mutation encompassing almost the whole C7 gene from exon 1 to exon 17 (c.1-?_2350+?del). A haplotype analysis showed that the large deletion mutation was inherited from the patient's father. To the best of our knowledge, this is the first confirmed case of C7 deficiency in Korea. PMID- 15831991 TI - Seroprevalence of Mycoplasma pneumoniae and Chlamydia pneumoniae in stable asthma and chronic obstructive pulmonary disease. AB - Mycoplasma pneumoniae and Chlamydia pneumoniae have been suggested to take part in the acute exacerbation of bronchial asthma and chronic obstructive pulmonary disease (COPD). Several studies have questioned whether they may play pathogenic roles in connection with bronchial asthma and COPD. This study was designed to evaluate the seroprevalences of M. pneumoniae and C. pneumoniae in stable asthma and COPD patients, and to compare with control patients. The medical records of one hundred forty patients who underwent M. pneumoniae and C. pneumoniae serology were retrospectively reviewed. Seroprevalences of M. pneumoniae and C. pneumoniae in the asthma group (11.1% and 8.3%, respectively) were higher than in the control group (4.4% and 2.2%, respectively) without statistical significance. The seroprevalence of M. pneumoniae in the COPD group (16.9%) was significantly higher than in the control group, and the seroprevalence of C. pneumoniae in the COPD group (3.4%) was higher than in the control group without statistical significance. This study raises important questions about the relation of M. pneumoniae and C. pneumoniae infection with stable asthma or COPD. PMID- 15831992 TI - Prognosis after surgical resection of M1a/M1b esophageal squamous cell carcinoma. AB - This study was undertaken to examine prognosis after resection for M1 disease in squamous cell esophageal carcinoma. Fifty-six patients with M1 esophageal cancer underwent esophageal resection with two or three-field nodal dissection from 1994 to 2001. Operative mortality occurred in 3 patients. Primary tumor sites were as follows; 10 upper, 23 middle, and 20 lower thoracic esophagus. They were found to have M1 disease by pathologic examination of dissected nodes, 24 M1a and 29 M1b. Forty-two patients (79%) were considered to have undergone curative resection. Chemotherapy and/or radiation therapy was given to 38 patients perioperatively. Recurrence was identified in 35 patients (66%) during a mean follow-up of 23 months. Overall median and 5-yr survivals were 19 months and 12.7%. Five-year survivals for M1a and M1b disease were 23.9% and 6.1%, respectively (p=0.0488). Curative resection tended to show better survival (p=0.3846). Chemotherapy and/or radiation therapy provided no advantage (p=0.5370). Multivariate analysis showed that M1b was significant risk factor over M1a disease. Our conclusion is that surgical resection can provide acceptable survival in thoracic squamous esophageal cancer with M1a disease. Survival differences between M1a and M1b disease support the current subclassification staging system. PMID- 15831993 TI - Effects of preoperative anxiety on gastric fluid acidity and volume. AB - The aim of this study was to evaluate the effect of preoperative anxiety on the gastric pH and volume. We studied 96 female patients aged 16-60 yr who underwent elective gynecological surgery. We classified the subjects into 2 groups, those presenting preoperative anxiety scores using visual analogue scale (VAS, 0-10) less than 5 (L-group, n=59), and those with 5 and more (H-group, n=37). Immediately after tracheal intubation, gastric contents were aspirated using a 14 F multiorifice nasogastric tube. The gastric acidity and volume of the two groups were not statistically different. Mean pH were 3.0 +/-1.8 and 3.0 +/-2.0 in each group (L-group and H-group) and mean gastric volume (mL) were 15.3 +/-11.7 and 11.8 +/-11.8, respectively. Nine (15.3%) patients in the L-group were considered to be 'at risk i.e. gastric pH <2.5 and volume >25 mL' and one patient (2.7%) in the H-group (p <0.05). The mean serum gastrin concentrations in both groups were similar (21.6 +/-9.8 vs. 20.2 +/-11.0 pg/mL). The pH and volume of preoperative gastric contents were not correlated with the preoperative anxiety. The results suggest that a low level of preoperative anxiety can be considered a risk factor for aspiration pneumonitis. PMID- 15831994 TI - DNA hypermethylation of tumor-related genes in gastric carcinoma. AB - The hypermethylation of the CpG islands is a common mechanism for the inactivation of tumor-related genes. In the present study, we analyzed the methylation status of genes for cell repair such as hMLH1, MGMT, and GSTP1, and a gastric cancer-specifically methylated DNA fragment, MINT 25 in gastric cancer cases and control groups. The study population consisted of 100 gastric cancer patients (50 distal and 50 proximal carcinomas), and 238 healthy controls. All genes showed more frequent hypermethylation in the cases than in the control group (p<0.0001). We investigated the association between promoter hypermethylation and relevant parameters including age, gender, alcohol consumption, smoking, and family history. There was a common hypermethylation of hMLH1 (p=0.008), MGMT (p= 0.0001), and GSTP1 (p=0.0003) in females. This study also demonstrates that hypermethylation was strongly associated with non-drinkers (MGMT, p=0.046 and MINT 25, p=0.049) and non-smokers (hMLH1, p=0.044; MGMT, p=0.0003; MINT 25, p=0.029). Moreover, the frequency of MINT 25 hypermethylation increased with age (p=0.037), and MGMT methylation was frequently detected in distal gastric cancer than in proximal type (p=0.038). Our study suggested that promoter hypermethylation of the genes involved in cell repair system and MINT 25 is associated strongly with some subgroups of primary gastric carcinoma. PMID- 15831995 TI - Promoter methylation of E-cadherin in hepatocellular carcinomas and dysplastic nodules. AB - In order to clarify the significance of E-cadherin methylation in multistep hepatocarcinogenesis, we examined the methylation status of the E-cadherin promoter region, using methylation-specific polymerase chain reaction in 64 hepatocellular carcinomas (HCCs) and 13 dysplastic nodules (DNs), and correlated these results with E-cadherin protein expression and clinicopathologic factors of HCCs. Promoter methylation was detected in 1 of 13 (7.7%) DNs, in 5 of 13 (38.5%) Edmondson and Steiner grade I HCCs, and in 27 of 51 (52.9%) grade II or III HCCs, and a significant correlation was observed between the methylation status and the stepwise progression of hepatocarcinogenesis (p=0.004). Reduced E-cadherin immunoreactivity was found in 18 of 64 (28%) HCCs, but in none of DNs. E-cadherin methylation status in HCCs was significantly correlated with microvascular invasion (p=0.02) and tumor recurrence (p=0.04), but not with reduced E-cadherin immunoreactivity. The Kaplan-Meier method showed that methylation status did not have a significant influence on the recurrence-free survival of HCC patients (p=0.15). Our results indicate that methylation of the E-cadherin promoter region is a frequent event in HCC, which may play an important role in the stepwise progression of hepatocarcinogenesis. And the promoter methylation of E-cadherin in HCC was found to be significantly correlated with microvascular invasion and recurrence. PMID- 15831996 TI - Angiotensin II AT1 receptor blockade changes expression of renal sodium transporters in rats with chronic renal failure. AB - We aimed to examine the effects of angiotensin II AT(1) receptor blocker on the expression of major renal sodium transporters and aquaporin-2 (AQP2) in rats with chronic renal failure (CRF). During 2 wks after 5/6 nephrectomy or sham operation, both CRF rats (n=10) and sham-operated control rats (n=7) received a fixed amount of low sodium diet and had free access to water. CRF rats (n=10) were divided into two groups which were either candesartan-treated (CRF-C, n=4) or vehicle-treated (CRF-V, n=6). Both CRF-C and CRF-V demonstrated azotemia, decreased GFR, polyuria, and decreased urine osmolality compared with sham operated rats. When compared with CRF-V, CRF-C was associated with significantly higher BUN levels and lower remnant kidney weight. Semiquantitative immunoblotting demonstrated decreased AQP2 expression in both CRF-C (54% of control levels) and CRF-V (57%), whereas BSC-1 expression was increased in both CRF groups. Particularly, CRF-C was associated with higher BSC-1 expression (611%) compared with CRF-V (289%). In contrast, the expression of NHE3 (25%) and TSC (27%) was decreased in CRF-C, whereas no changes were observed in CRF-V. In conclusion, 1) candesartan treatment in an early phase of CRF is associated with decreased renal hypertrophy and increased BUN level; 2) decreased AQP2 level in CRF is likely to play a role in the decreased urine concentration, and the downregulation is not altered in response to candesartan treatment; 3) candesartan treatment decreases NHE3 and TSC expression; and 4) an increase of BSC-1 is prominent in candesartan-treated CRF rats, which could be associated with the increased delivery of sodium and water to the thick ascending limb. PMID- 15831997 TI - M3 subtype of muscarinic receptors mediate Ca2+ release from intracellular stores in rat prostate neuroendocrine cells. AB - Our previous studies document the expression of adrenoceptors and purinoceptors in the rat prostate neuroendocrine cells (RPNECs). However, a direct investigation of the receptors for acetylcholine (ACh) is still lacking in the prostate neuroendocrine cells. RPNECs were freshly isolated from the ventral lobes of rat prostate by using collagenase. Effects of ACh and various muscarinic antagonists on the intracellular Ca(2+) concentration ([Ca(2+)](c)) were investigated by using the fura-2 spectrofluorimetry. Single-cell RT-PCR analysis was applied to identify the transcripts for the muscarinic receptor subtypes. ACh (5 microM) induced a sharp transient increase in the [Ca(2+)](c) of RPNECs, which was independent of the extracellular Ca(2+). In the same RPNECs, high KCl (60 mM), phenylephrine (5 microM), UTP (P2Y(1/2) agonist, 50 microM), and alpha, beta meATP (P2X(1/3) agonist, 0.5 microM) also increased the [Ca(2+)](c). The ACh induced [Ca(2+)](c) change (delta [Ca(2+)](c)) was blocked by atropine or by para fluorohexahydrosiladifenidol (M(3) antagonist, 0.3 microM), but not by telenzepine (M(1) antagonist, 1 microM) and himbacine (M(2) and M(4) antagonist, 1 microM). The single-cell RT-PCR demonstrated the selective expression of mRNAs for M(3) in RPNECs. In summary, RPNECs express M(3) muscarinic receptors that are linked to the release of Ca(2+) from intracellular stores. The Ca(2+) signals of RPNECs might mediate the parasympathetic regulation of prostate gland. PMID- 15831998 TI - Nomograms for the prediction of pathologic stage of clinically localized prostate cancer in Korean men. AB - We analyzed the prostate cancer data of 317 Korean men with clinically localized prostate cancer who underwent radical prostatectomy at Asan Medical Center between June 1990 and November 2003 to construct nomograms predicting the pathologic stage of these tumors, and compared the outcome with preexisting nomograms. Multinomial log-linear regression was performed for the simultaneous prediction of organ-confined disease (OCD), extracapsular extension (ECE), seminal vesicle invasion (SVI) and lymph node metastasis (LNM) using serum prostate-specific antigen (PSA), Gleason score and clinical stage. Nomograms representing percent probabilities were constructed and compared with those presented by Partin et al. by calculating areas under the receiver operating characteristics (ROC) curves. Median serum PSA at surgery was 10.8 ng/mL, and median biopsy Gleason score was 7. Overall OCD, ECE, SVI and LNM rates were 59.6%, 20.5%, 11.7% and 8.2%, respectively, and areas under the curves were 0.724, 0.626, 0.662, and 0.794, respectively. Pathologic stage of localized prostate cancer in Korean men may be predicted using the Partin table, with acceptable accuracy for OCD and LNM, but less so for ECE and SVI. PMID- 15831999 TI - Antioxidant vitamins and lipid peroxidation in patients with cervical intraepithelial neoplasia. AB - The purpose of this study was to investigate the implications of dietary intake and the level of plasma antioxidant, lipid peroxidation, and antioxidant capacity in Korean women with cervical intraepithelial neoplasia (CIN). From October 2002 to March 2003, 58 patients diagnosed with CIN (confirmed with colposcopy directed biopsy) and 86 patients without any cervical disease as control group were enrolled in the study at the Department of Gynecology cancer center at Samsung Cheil Hospital. The intake of antioxidant vitamins in both groups exceeded the amount recommended by the Korea RDA, 7th edition. The plasma concentration of Vitamin C was significantly lower in the CIN group (0.36 mg/dL) than in the control group (0.48 mg/dL) (p<0.05). The two groups showed similar plasma concentrations of beta-carotene, alpha-tocopherol, and retinol. The average concentration of malondialdehydes in the CIN group, 7.23 mmol/mL, was significantly higher than in the control group, 5.18 mmol/mL (p<0.01). The total radical trapping antioxidant potential concentration of plasma was significantly higher in the CIN group (1.15 mM) than in the control group (1.25 mM) (p<0.05). These results suggest that there is a possible correlation between cervical intraepithelial neoplastic processes and changes in the plasma antioxidative system. PMID- 15832000 TI - Short-term effects of 308-nm xenon-chloride excimer laser and narrow-band ultraviolet B in the treatment of vitiligo: a comparative study. AB - We compared the clinical efficacy of a short-term intervention of 308-nm excimer laser with that of narrow-band UVB (NBUVB) phototherapy for vitiligo patients to see the early response. Twenty-three symmetrically patterned patches of vitiligo on 8 patients were selected. Vitiligo patches on one side of the body were treated 2 times per week for a maximum of 20 treatments with the excimer laser, and NBUVB phototherapy was used on patches on the other side. Improvement (repigmentation) was assessed on a visual scale via serial photographs taken every five treatments and scored as follows: 0,< or =1% improvement; 1,< or =25% improvement; 2, 26-50% improvement; 3, 51-75% improvement; and 4, > or =75% improvement. At five treatments, the excimer laser-treated patches had an average score of 0.26, compared with 0.04 for patches treated with NBUVB phototherapy. A slightly higher repigmentation (p>0.05) in the excimer treated area was thus observed. At 10, 15, or 20 treatments, the differences between the average scores were significant: 0.83, 1.17, and 1.39 for the excimer-treated patches, and 0.17, 0.30, and 0.74 for the NBUVB phototherapy-treated areas (p<0.05). In conclusion, the 308-nm excimer laser appears to be more effective than NBUVB phototherapy, as it produces more rapid and profound repigmentation. PMID- 15832001 TI - Primary facial skin cancer: clinical characteristics and surgical outcome in Chungbuk Province, Korea. AB - Clinical features of facial skin cancer in Asian population including Korean are not readily available. In the present study, we analyzed the clinical characteristics and the surgical results of primary facial skin cancer in Chungbuk Province, Korea. Eighty-six cases of primary facial skin cancer collected during a 10-yr period (1994-2003) were retrospectively reviewed about the clinical characteristics including age, sex, annual diagnostic rate, types of tumor, specific sites of occurrence, and the surgical results. The average age at the diagnosis was 67 and male to female ratio was 1 to 1.05. The average annual diagnostic rate was 0.73% and the rate surged during the period 2001-2003 compared with the period 1994 to 2000. Basal cell carcinoma was the most common tumor and the nose was the most frequent site. Traditional surgical excision with immediate reconstruction was performed in 81 cases. During the 23 months of average follow-up, three patients had recurrences (3.7%) and three patients had secondary skin cancers. Facial skin cancer is increasing in the province and basal cell carcinoma is most frequent. Traditional surgical excision and immediate reconstruction with local flap are a good therapeutic modality with an acceptable recurrence rate. PMID- 15832002 TI - Cardiac dysrhythmias,cardiomyopathy and muscular dystrophy in patients with Emery Dreifuss muscular dystrophy and limb-girdle muscular dystrophy type 1B. AB - Emery-Dreifuss muscular dystrophy (EDMD) and limb-girdle muscular dystrophy type 1B (LGMD1B) are characterized by cardiac dysrhythmias, late-onset cardiomyopathy, slowly progressive skeletal myopathy and contractures of the neck, elbows and ankles. The causative mutation is either in the emerin gene (X-linked recessive EDMD) or lamin A/C gene (autosomal dominant EDMD2 or LGMD1B). We report three cases of EDMD, EDMD2 and LGMD1B. A 14-yr-old boy showed limitation of cervical flexion and contractures of both elbows and ankles. Sinus arrest with junctional escape beats was noted. He was diagnosed as X-linked recessive EDMD (MIM 310300). A 28-yr-old female showed severe wasting and weakness of humeroperoneal muscles. Marked limitation of cervical flexion and contractures of both elbows and ankles were noted. Varying degrees of AV block were noted. She was diagnosed as autosomal dominant EDMD2 (MIM 181350). A 41-yr-old female had contractures of both ankles and limb-girdle type muscular dystrophy. ECG revealed atrial tachycardia with high grade AV block. She was diagnosed as autosomal dominant LGMD1B (MIM 159001). Cardiac dysrhythmias in EDMD and LGMD1B include AV block, bradycardia, atrial tachycardia, atrial fibrillation, and atrial standstill, causing sudden death necessitating pacemaker implantation. Cardiologists should know about these unusual genetic diseases with conduction defects, especially in young adults. PMID- 15832003 TI - Neuroprotective effect of ginseng total saponins in experimental traumatic brain injury. AB - In the present study, we investigated whether ginseng total saponins (GTSs) protect hippocampal neurons after experimental traumatic brain injury (TBI) in rats. A moderate-grade TBI was made with the aid of a controlled cortical impact (CCI) device set at a velocity of 3.0 m/sec, a deformation of 3.0 mm, and a compression time of 0.2 sec at the right parietal area for adult male Sprague Dawley rats. Shamoperated rats that underwent craniectomy without impact served as controls. GTSs (100 and 200 mg/kg) or saline was injected intraperitoneally into the rats immediately post-injury. Twenty-four hours after the injury, the rats underwent neurological evaluation. Contusion volume and the number of hippocampal neurons were calculated with apoptosis evaluated by TUNEL staining. 24 hr post-injury, saline-injected rats showed a significant loss of neuronal cells in the CA2 region of the right hippocampus (53.4%, p<0.05) and CA3 (34.6%, p<0.05) compared with contralateral hippocampal region, a significant increase in contusion volume (34+/-8 microL), and significant increase in neurologic deficits compared with the GTSs groups. Treating rats with GTSs seemed to protect the CCI induced neuronal loss in the hippocampus, decrease cortical contusion volume, and improve neurological deficits. PMID- 15832004 TI - Biodegradable polymer releasing antibiotic developed for drainage catheter of cerebrospinal fluid: in vitro results. AB - The authors developed a biodegradable polymer that releases an antibiotic (nalidixic acid) slowly and continuously, for prevention of catheter-induced infection during drainage of cerebrospinal fluid. We investigated the in vitro antibiotic releasing characteristics and bacterial killing effects of the new polymer against E. coli. The novel fluoroquinolone polymer was prepared using diisopropylcarbodiimide, poly (e-capro-lactone) diol, and nalidixic acid. FT-IR, mass spectrometry, and elemental analysis proved that the novel antibacterial polymer was prepared successfully without any side products. Negative MS showed that the released drug has a similar molecular weight (M.W.=232, 350) to pure drug (M.W.=232). In high pressure liquid chromatography, the released drug and drug-oligomer showed similar retention times (about 4.5-5 min) in comparison to pure drug (4.5 min). The released nalidixic acid and nalidixic acid derivatives have antibacterial characteristics against E. Coli, Staphylococcus aureus, and Salmonella typhi, of more than 3 months duration. This study suggests the possibility of applying this new polymer to manufacture drainage catheters that resist catheter-induced infection, by delivering antibiotics for a longer period of more than 1 month. PMID- 15832005 TI - Factor structure of the Korean version of illness intrusiveness rating scale: cross-cultural implications. AB - The Illness Intrusiveness Rating Scale (IIRS) measures illness-induced disruptions to 13 domains of lifestyles, activities, and interests. A stable three-factor structure has been well documented; however, the cross-cultural validity of this scale needs to be tested. This study investigated the factor structure of the Korean version of IIRS in 712 outpatients at a university medical center. A predominant diagnosis of the patients was rheumatoid arthritis (47%). The Center for Epidemiological Studies-Depression Scale (CES-D), and Health Assessment Questionnaire (HAQ) were also administered. Exploratory Principal Component Analysis identified a two-factor structure, "Relationships and Personal Development (RPD)" and "Instrumental", accounting for 57% of the variance. Confirmatory analyses extracted an identical factor structure. However, a goodness-of-the fit test failed to support two-factor solution (chi(2)=138.2, df=43, p<.001). Two factors had high internal consistency (RPD, alpha=.89; Instrumental, alpha=.75) and significantly correlated with scores of HAQ (RPD, r=.53, p<.001; Instrumental, .r=44, p<.001) and CES-D (RPD, .r=55, p<.001; Instrumental, .r=43, p<.001). These findings supported construct validity of the Korean version of IIRS, but did not support cross-cultural equivalence of the factor structure. PMID- 15832006 TI - Antinociceptive interactions between intrathecal gabapentin and MK801 or NBQX in rat formalin test. AB - Antagonists for spinal N-methyl-D-aspartate (NMDA) and amino-hydroxy-methtyl isoxazolepropionate (AMPA) receptors are effective in attenuating acute nociception or injury-induced hyperalgesia. The antinociception of spinal gabapentin is developed in injury-induced hyperalgesia without affecting acute nociception. The authors evaluated the effects of intrathecal gabapentin, NMDA antagonist (MK801) and AMPA antagonist (NBQX) in the formalin test which shows injury-induced hyperalgesia as well as acute pain. We further assessed the interactions between gabapentin and either MK801 or NBQX. Male Sprague-Dawley rats were implanted with intrathecal catheters. To evoke pain, 50 microL of 5% formalin solution was injected into the hindpaw. The interaction was investigated by a fixed dose analysis or an isobolographic analysis. MK801 and NBQX suppressed flinching responses during phase 1 of the formalin test, while gabapentin had little effect on phase 1. All three agents decreased the phase 2 flinching response. A fixed dose analysis in phase 1 showed that gabapentin potentiated the antinociceptive effect of MK801 and NBQX. Isobolographic analysis in phase 2 revealed a synergistic interaction after coadministration of gabapentin-MK801 or gabapentin-NBQX. Correspondingly, spinal gabapentin with NMDA or AMPA antagonist may be useful in managing acute pain and injury-induced hyperalgesia. PMID- 15832007 TI - Splenic abscess associated with endocarditis in a patient on hemodialysis: a case report. AB - Splenic abscess is an unusual condition usually seen in immunocompromised patients or associated with intravenous drug abuses. Several conditions including trauma, immunodeficiency, corticosteroid and/or immunosuppressive therapy and diabetes mellitus have been listed under the predisposing factors for a splenic abscess. Splenic abscess in a patient on hemodialysis is a rare but life threatening condition if not corrected. We describe a case of splenic abscess with bacterial endocarditis on maintenance hemodialysis. He had staphylococcal septicemia secondary to bacterial endocarditis at the mitral valve from the dialysis access-site infection. Although hematologic seeding from endocarditis has been the predisposing factor for splenic abscess, we postulate that access site infections may predispose hemodialysis patients to splenic abscess. Splenic abscess may be considered as one of the causes when patients on hemodialysis develop unexplained fever. PMID- 15832008 TI - Preoperative embolization in surgical treatment of a primary hemangiopericytoma of the rib: a case report. AB - Primary hemangiopericytoma of the rib is extremely rare and only a few cases have been reported. A 62-yr-old man presented with an aching chest pain and dyspnea. Thoracic computed tomography revealed a homogenous mass expanding the right seventh rib. A diagnosis of hemangiopericytoma was established by percutaneous needle biopsy. Preoperative embolization of the feeding vessels of the tumor was performed in order to prevent perioperative bleeding. There was no significant bleeding during the surgery, where complete resection of the tumor with 7th to 9th ribs with a surgical margin of 5 cm was performed. Postoperative course was uneventful and there has been no recurrence for thirteen months. To our knowledge, there has been no report to apply a preoperative embolization of a primary hemangiopericytoma of the rib. PMID- 15832009 TI - CD4+CD56+ lineage negative hematopoietic neoplasm: so called blastic NK cell lymphoma. AB - Blastic natural killer (NK) cell lymphoma is a rare neoplasm characterized by blastoid tumor cells expressing CD4 and CD56, with predominant skin involvement. Although this tumor has been regarded as a neoplasm related to NK cell, recent studies suggested that it is derived from plasmacytoid dendritic cells, but not from NK cell. Herein we report 4 cases of CD4+CD56+ lineage marker-blastic NK cell lymphomas with a review of literatures. The patients were 3 men and one woman. Three of them were young (17, 18, and 22 yr old). Three patients had skin lesions, at initial presentation in two patients and during the course of disease in other patient. Histologically, tumors consisted of monotonous medium to large blastoid cells showing no necrosis, angiocentric growth or epidermotrophism. All four tumors were CD4+ and CD56+. Three expressed CD68 antigen. Lineage specific markers for B- and T cell were negative. All tumors did not express myeloperoxidase. T-cell receptor gene rearrangement, EBV, CD13 and CD33 were negative. In one patient, tumor cells arranged in Homer-Wright type pseudorosette and expressed terminal deoxynucleotidyl transferase(TdT). Despite the standard lymphoma chemotherapy, the tumors, except one lost during follow-up, progressed and relapsed. The patients died 8-60 months after diagnosis. PMID- 15832010 TI - A case of multiple mucosa-associated lymphoid tissue (MALT) lymphoma of the colon identified as simple mucosal discoloration. AB - Most colonic multiple mucosa-associated lymphoid tissue (MALT) lymphomas are confirmed with a histologic and immunohistochemical staining of the mucosal biopsy specimen obtained during colonoscopic examinations. Endoscopically, colonic MALT lymphomas frequently appear as protruding and/or ulcerative lesions, and there are not so many reports of colonic MALT lymphoma as compared to the frequent reports of MALT lymphoma of stomach. We report a unique case of colonic MALT lymphoma presenting as a simple reddish discoloration of mucosa; this presentation has never been describe before. Our patient was a 47-yr-old male who suffered from tenesmus and mucoid stool. A colonoscopy was accomplished, followed by a histologic examination and we diagnosed a colonic MALT lymphoma. Staging of the disease was done because this was necessary for choosing the modality of treatments. The patient was then treated with polychemotherapy in conjunction with radiation therapy. PMID- 15832011 TI - Harlequin syndrome with crossed sympathetic deficit of the face and arm. AB - Harlequin syndrome is characterized by unilateral hyperhidrosis and flushing, which are predominantly induced by heat or exercise. Usually, the sympathetic deficits confine to the face. Rarely, the autonomic deficits involve the arm or the parasympathetic neurons in the ciliary ganglia. We report a 43-yr-old woman who presented with facial flushing and sweating in the right side, which were mainly induced by exercise. The facial flushing accompanied relative coldness in the right arm. Valsalva maneuver, cold pressure and 0.125% pilocarpine test, and computed tomography of the chest were normal. The crossed sympathetic deficit in the left face and right arm suggested that the lesions were multifocal. The sympathetic impairment in our patient may lie on a spectrum of pre- and postganglionic autonomic dysfunction, which was observed in Holmes-Adie, Ross, and Guillain-Barre syndrome. PMID- 15832012 TI - The poor outcome of the delayed diagnosis of acute spontaneous spinal epidural hematoma: two cases report. AB - We present two patients who had acute paraplegia with sensory loss due to spontaneous spinal epidural hematoma (SSEH). One had myocardial infraction and the other had deep vein thrombosis, and the former was treated with anticoagulants and the latter was treated with thrombolytic agent. We analyzed the neurological status of our two cases each between its preoperative and postoperative state. Postoperatively both showed no improvement of neurologic symptom, and on follow-up of 12 months, one showed no neurologic improvement and the other showed a insignificant improvement of lower extremity muscle power (trace knee extensor/ankle dorsi-flexor). We thought that this poor outcome was due to delayed operation, which was done more than 24 hr after the symptom onset. The outcome in SSEH is essentially determined by the time taken from symptom onset to operation. Therefore, early and precise diagnosis such as careful history taking and MRI evaluation is necessary. PMID- 15832013 TI - Neuroendoscopic removal of large choroid plexus cyst: a case report. AB - Choroid plexus cysts (CPCs) are the most common neuroepithelial cysts, occurring in more than 50% of some autopsy series. They are typically small and asymptomatic and are discovered incidentally in older patients, usually in the trigone of the lateral ventricle. Symptomatic CPCs (usually exceptionally large, 2-8 cm) are rare. The authors report a case of large symptomatic choroid plexus cyst, located in the trigone of the right lateral ventricle in a 26-yr-old man who presented with headache and vomiting. The patient underwent endoscopic removal through a burr hole placed 3 cm from the midline and just behind the hair line. The histological examination of the cyst wall was consistent with choroid epithelium. Despite of postoperative intraventricular hemorrhage and catheter infection, he discharged home without neurologic deficits. The endoscopic fenestration rather than excision should be considered as the first surgical procedure because the goal of treatment is shrinkage of the cyst until normal cerebrospinal fluid flow is restored. PMID- 15832014 TI - Analysis of loss of heterozygosity in Korean patients with keratoacanthoma. AB - Loss of heterozygosity (LOH) has been established as an important genetic mechanism giving rise to malignant neoplasia. The mechanism of LOH has been shown to cause basal cell carcinoma and malignant melanoma as well as other types of skin cancer. A few studies on LOH in sporadic keratoacanthomas have been reported. The purpose of this study was to investigate the significance of LOH in the pathogenesis of sporadic keratoacanthomas developed in 10 Korean patients. The presents of LOH at 7 microsatellite markers (D2S286, D3S1317, D5S346, D9S160, D9S171, D10S185, and D17S261) were evaluated in sporadic keratoacanthomas. LOH was found in only 1 of 10 cases at D10S185. The low frequency of LOH detected in this study suggests that LOH may not be significant in the induction of sporadic keratoacanthomas. PMID- 15832015 TI - Sketches of otohistory. Part 8: The emergence of vestibular science. PMID- 15832017 TI - Accumulation of metals and minerals from phosphate binders. AB - Metals and minerals that depend on renal clearance may accumulate to toxic levels in patients with marginal kidney function. Toxicities of aluminum-based phosphate binders became apparent approximately 25 years ago. Nephrologists now recognize cardiovascular calcification may follow use of calcium-based phosphate binders. Five lessons can be learned: (1)safety must not be assumed in absence of data; (2) all evidence for causal linkage of toxicities from therapeutics must be considered, including animal data; (3) clinical trials are unlikely to reveal the spectrum of problems from long-term drug exposure; (4) complications can remain unrecognized until late in post-introduction surveillance; (5) minerals important for normal function can be toxic with excess accumulation. Introduction of new agents necessitates caution - it is difficult to change practice once a therapeutic is commonplace. Lessons learned about hazards of past phosphate binders must be applied judiciously when evaluating long-term risks/safety of novel metal-based binders such as lanthanum carbonate. PMID- 15832018 TI - Calcium loading, calcium accumulation, and associated cardiovascular risks in dialysis patients. AB - Calcium and phosphate imbalances are important mutable risk factors for cardiovascular disease in chronic kidney disease (CKD). Nearly all dialysis patients require phosphate binders. These include traditional calcium-based compounds and, more recently, the calcium-free, metal-free, non-absorbed agent, sevelamer hydrochloride. Both binder types reduce serum phosphorus, but differ with respect to calcium load and metabolism. Absorption from calcium-based agents very likely promotes positive total calcium balance in many patients. Positive calcium balance is inappropriate in adults and may promote or accelerate soft tissue and vascular calcification even in the absence of hypercalcemia. Calcium accumulation in heart and vascular tissues contributes to rapidly progressive cardiovascular calcification - a strong predictor of cardiovascular and all-cause mortality in stage 5 CKD. More than two-thirds of stage 5 CKD patients have calcification scores above the 75th percentile for matched controls -- scores associated with extremely high risk of cardiovascular events and death. PMID- 15832019 TI - Cardiovascular calcification progression -- a comparison of sevelamer and calcium based phosphate binders. PMID- 15832020 TI - Spinal grey matter infarction after aortic surgery: a case of persistent pure flaccid paraplegia. PMID- 15832021 TI - Emergent stenting in carotid artery dissection: MRI-guided therapeutic decision making. PMID- 15832022 TI - An unusual vascular cause of Horner's syndrome. PMID- 15832023 TI - Co-occurrence of CADASIL and isolated CNS angiitis. PMID- 15832024 TI - Bilateral top of carotid occlusion presenting as basilar thrombosis. PMID- 15832025 TI - Isolated type 2 diabetes mellitus causes myocardial dysfunction that becomes worse in the presence of cardiovascular diseases: results of the myocardial doppler in diabetes (MYDID) study 1. AB - AIMS: Patients with type 2 diabetes mellitus (DM) often suffer disproportionately and have a worse outcome when burdened with cardiovascular complications compared with those without DM. A specific heart muscle disease reportedly caused by DM per se may explain this. We sought to investigate whether an echo Doppler diagnosis of such a myocardial disease is clinically relevant in DM with or without coexistent coronary artery disease (CAD) and/or hypertension (HTN). SUBJECTS AND METHODS: Two hundred subjects (127 males, 73 females, 56 +/- 10 years) including controls (n = 23), patients with HTN (n = 20), CAD (n = 35), uncomplicated DM (n = 59), DM+HTN (n = 27), DM+CAD (n = 16) and DM+CAD+HTN (n = 20) underwent tissue Doppler-enhanced dobutamine stress echocardiography. Myocardial function was assessed by measuring left ventricular myocardial peak systolic velocity (PSV) and early diastolic velocity at rest and during peak stress, besides measurements of standard Doppler variables. RESULTS: Average left ventricular PSV at rest was significantly lower in CAD (4.7 +/- 1.5) compared with controls (5.7 +/- 1.2) and in DM+CAD+HTN (4.6 +/- 1.4) compared with DM (5.6 +/- 1.3; all p < 0.05). During peak stress, lower PSV persisted in CAD (9.5 +/- 3.1) and DM+CAD+HTN (8.1 +/- 2.7), while appearing de novo in DM (11.3 +/- 2.6) and HTN (11.0 +/- 2.3) unlike in the controls (12.5 +/- 2.5; all p < 0.001). When pooled together, DM subjects with CAD and/or HTN or both had significantly lower PSV (9.1 +/- 2.7) than those without (10.0 +/- 2.8; p < 0.001). Early diastolic velocity response was equally lower in both groups compared with the controls. CONCLUSION: The results suggest that dobutamine stress unmasks myocardial functional disturbances caused by uncomplicated DM. The discrete disturbances become quantitatively more pronounced in the presence of coexistent cardiovascular diseases. PMID- 15832026 TI - Intracoronary epinephrine for contrast-medium-induced microvascular obstruction in a chronically hemodialyzed patient. PMID- 15832027 TI - High-density lipoprotein and vascular tone in healthy subjects. PMID- 15832028 TI - Predictors of benefit in angina patients one year after completing enhanced external counterpulsation: initial responders to treatment versus nonresponders. AB - Enhanced external counterpulsation (EECP) has been shown to reduce Canadian Cardiovascular Society angina class. This study examines the factors that affect the reduction at 1 year, especially in patients who do not demonstrate an initial response. The data of 2,007 consecutive patients enrolled in the International EECP Patient Registry were analyzed. After 36.6 +/- 4.9 h of EECP, angina was reduced by at least one class in 82.7%. At 1 year, 35.4% of initial nonresponders and 70.6% of responders remained improved by at least one angina class and free of major adverse cardiovascular events. Multivariate predictors of 1-year benefit are initial response to treatment (odds ratio 4.5, 95% CI 3.5-5.8), baseline angina class compared with class IV (odds ratios: class I 2.1, CI 0.93-4.81; class II 0.62, CI 0.43-0.87; class III 0.80, CI 0.62-1.01) and no history of congestive heart failure (odds ratio 1.41, CI 1.14-1.74). PMID- 15832029 TI - Heat shock protein 70 and tumor necrosis factor alpha in Taiwanese patients with dementia. AB - This study was to determine whether polymorphisms of heat shock protein 70-1 (HSP70-1) and tumor necrosis factor alpha (TNF-alpha) are associated with the risk of Alzheimer's disease (AD) and vascular dementia (VaD). Using the criteria of the NINCDS-ADRDA and NINDS-AIREN, 125 AD patients, 57 VaD patients and 109 ethnically matched nondemented controls were enrolled. The HSP70-1 -110 A/C and TNF-alpha -1031 T/C, -863 C/A and -857 C/T polymorphisms were analyzed by means of genotype or haplotype association methods. None of the four genotypes examined showed a statistically significant difference in genotype distribution between the AD cases and controls. However, the HSP70-1 -110 CC genotype occurred more frequently among AD cases (p=0.0821; odds ratio: 2.08; 95% confidence interval, CI: 0.92-4.98). The overall genotype distribution among the VaD cases tended to be different at the HSP70-1 -110 and TNF-alpha -1031 sites (p=0.0604 and 0.0316, respectively). The HSP70-1 -110 CC genotype was more frequent (p=0.0459), and the association of the -110 CC genotype with VaD was evident (p=0.0207; odds ratio: 3.22; 95% CI: 1.20-8.87). The more frequent TNF-alpha -1031 TC genotype (p=0.0614) was also evidently associated with VaD (p=0.0209; odds ratio: 2.32; 95% CI: 1.14-4.78). Multivariate analysis demonstrated the synergistic effect of the HSP70-1 -110 CC and TNF-alpha -1031 TC/CC genotypes on VaD (p=0.0091; odds ratio: 10.09; 95% CI: 2.01-75.97). Haplotype analysis among TNF-alpha -1031, 863, -857 sites revealed that -1031C-857C may act as a risk haplotype among VaD cases (p=0.0132, odds ratio: 2.26; 95% CI: 1.19-4.33). Our results suggest a potential protective role for HSP70 in both VaD and AD, whereas TNF-alpha may act as a risk factor only for VaD, and not for AD. PMID- 15832030 TI - Cognitive reserve: a SPECT study of 132 Alzheimer's disease patients with an education range of 0-19 years. AB - This study examines the associations between education, cerebral perfusion, and cognitive test performance among 132 patients with Alzheimer's disease. The participants had had between 0 and 19 years of formal schooling, and had either mild or moderate dementia according to the Clinical Dementia Rating Scale. Cerebral perfusion was evaluated by the (99m)Tc-hexamethylpropylene amine oxime single photon emission computed tomography. The Mini-Mental State Examination and the Cognitive Abilities Screening Instrument were used to assess cognitive performance. For patients at each clinical dementia severity level, statistical parametric mapping was used to examine voxel by voxel the association between education and cerebral perfusion, and Pearson's correlation coefficients were calculated between education and cognitive test scores. Years of formal schooling had negative associations with cerebral perfusion and positive associations with cognitive test scores. The brain regions showing a significant education effect on perfusion involved bilateral posterior association areas in mild dementia, and bilateral parieto-temporo-frontal areas in moderate dementia. The present findings indicate that the cognitive reserve effect starts at the low end of the education range. They also suggest that the main effect of more education is a more facile use of alternative brain circuits instead of locally increased synaptic connections. PMID- 15832031 TI - Do vascular lesions and related risk factors influence responsiveness to donepezil chloride in patients with Alzheimer's disease? AB - The purpose was to identify vascular influences on the responsiveness to donepezil chloride. The study included 50 untreated probable Alzheimer's disease patients with the Modified Hachinski Ischemic Score <4. We assessed baseline cognitive status using the Revised Hasegawa Dementia Scale (HDS-R), Clinical Dementia Rating (CDR) and the clock drawing test (CDT). The response to 5 mg of donepezil was monitored by the CDT for 12 months. Patients were classified as true responders (TR), unchanged (UC) and non-responders according to changes on the CDT in response to treatment. High HDS-R scores, low CDT scores, low CDR and presence of hypertension (HBP) and periventricular hyperintensities (PVH) predicted a TR- or UC-type outcome. Aggravation of executive function by HBP and/or PVH and its improvement by donepezil may have been detected by the CDT. PMID- 15832032 TI - Differentiation of dementia with Lewy bodies from Alzheimer's disease using brain SPECT. AB - We compared regional cerebral blood flow (CBF) patterns in patients with dementia with Lewy bodies (DLB) and Alzheimer's disease (AD) using single photon emission computed tomography (SPECT) and investigated the diagnostic utility of SPECT study in differentiating between DLB and AD. SPECT data on 20 patients with DLB and 75 patients with AD were analyzed using three-dimensional stereotactic surface projections. Regional CBF reduction was determined by quantitative analysis using stereotactic extraction estimation method. The DLB group showed a significant CBF reduction in the temporoparietal, frontal lobe and posterior cingulate, similar to the CBF pattern in the AD group, but regional CBF in the medial and lateral occipital lobes decreased significantly in patients with DLB compared with patients with AD. Receiver operating characteristic analysis revealed that regional CBF measurement of the medial occipital lobe, including the cuneus and lingual gyrus, yielded a sensitivity of 85% and a specificity of 85% in discriminating DLB from AD. Objective and quantitative CBF measurement in the medial occipital lobe may be useful in the clinical differentiation of DLB and AD. PMID- 15832033 TI - Evaluating associations between 5-HTTLPR polymorphism and Alzheimer's disease for Korean patients. AB - There has been growing evidence that the serotonin (5-HT) system is important in the regulation of memory and thus might be associated with Alzheimer's disease (AD), while research results on this issue have been inconsistent. The 5-HT system has also been suggested to be responsible for a significant portion of the behavioural aspects of AD. This study aimed to investigate the associations of the 5-HT transporter gene linked polymorphic region (5-HTTLPR) polymorphism with AD and delusional/aggressive symptoms of AD in Korean samples of 65 patients and 43 controls. The 5-HTTLPR polymorphism was neither associated with AD nor with delusional/aggressive symptoms of AD. It was suggested that phenotypic expression of the 5-HTTLPR polymorphism might be varied according to ethnic differences. PMID- 15832034 TI - Plasma homocysteine concentration and its relation to symptoms of vascular disease in psychogeriatric patients. AB - BACKGROUND: There is a high frequency of elevated plasma total homocysteine (tHcy) concentrations in elderly patients with mental disorders. Psychogeriatric patients with a history of vascular disease exhibit a significantly higher plasma tHcy concentration than patients without vascular disease. METHOD: The main reason for the present study is to further investigate the association between plasma tHcy concentration and vascular disease in psychogeriatric patients. We therefore investigated 152 psychogeriatric patients and determined plasma tHcy and its most important determinants (serum folate and serum cobalamin, serum cystatin C and serum creatinine). The patients were divided into two groups according to the presence of vascular disease. Eighty-seven patients had concomitant vascular disease. We also analysed the natriuretic peptide N-terminal pro brain natriuretic peptide (NT-proBNP) and protein S-100B in serum. NT-proBNP is a marker for congestive heart failure, whereas protein S-100B is a marker for brain damage. RESULTS: The plasma tHcy concentration is elevated in the presence of dementia or vascular disease in psychogeriatric patients. The presence of dementia or vascular disease is also associated with higher age, renal impairment and lower serum folate concentration than in patients without dementia or vascular disease. Furthermore, we observed elevated serum concentrations of NT proBNP in patients with dementia or vascular disease as a sign of poorer cardiovascular status. Likewise, protein S-100B concentrations were elevated in patients with dementia or vascular disease, possibly indicating brain damage in these groups of patients. CONCLUSION: The high frequency of comorbidity of vascular disease and mental illness indicates a possibility to prevent and treat psychogeriatric disease by actively counteracting vascular disease in patients with psychogeriatric symptoms. Routine determination of NT-proBNP is valuable for obtaining information about cardiovascular status. PMID- 15832035 TI - Test-retest reliability of the dementia rating scale-2: alternate form. AB - The purpose of this study was to examine the test-retest reliability of the newly developed Dementia Rating Scale-2: Alternate Form (DRS-2:AF) in a community dwelling sample of older adults. Participants were administered the DRS-2:AF during two separate testing sessions; the interval between sessions was between 12 and 28 days. The stability coefficient for the Total Score was quite high (0.93), and reliability coefficients for the subscale scores ranged from adequate to high. This project provides evidence for the test-retest reliability of the DRS-2:AF. Given the need for cognitive status measures with equivalent forms, the DRS-2:AF is recommended as a reliable tool in the assessment of dementia. PMID- 15832036 TI - Supplemental use of antioxidant vitamins and subsequent risk of cognitive decline and dementia. AB - There are conflicting reports about the potential role of vitamin antioxidants in preventing and/or slowing the progression of various forms of cognitive impairment including Alzheimer's disease (AD). We examined longitudinal data from the Canadian Study of Health and Aging, a population-based, prospective 5-year investigation of the epidemiology of dementia among Canadians aged 65+ years. Our primary objective was to examine the association between supplemental use of antioxidant vitamins and subsequent risk of significant cognitive decline (decrease in 3MS score of 10 points or more) among subjects with no evidence of dementia at baseline (n=894). We also explored the relationship between vitamin supplement use and incident vascular cognitive impairment (VCI; including a diagnosis of vascular dementia, possible AD with vascular components and VCI but not dementia), dementia (all cases) and AD. After adjusting for potential confounding factors assessed at baseline, subjects reporting a combined use of vitamin E and C supplements and/or multivitamin consumption at baseline were significantly less likely (adjusted OR 0.51; 95% CI 0.29-0.90) to experience significant cognitive decline during a 5-year follow-up period. Subjects reporting any antioxidant vitamin use at baseline also showed a significantly lower risk for incident VCI (adjusted OR 0.34, 95% CI 0.13-0.89). A reduced risk for incident dementia or AD was not observed. Our findings suggest a possible protective effect for antioxidant vitamins in relation to cognitive decline but randomized controlled trials are required for confirmation. PMID- 15832037 TI - Association between angiotensin-converting enzyme gene polymorphism and Alzheimer's disease in a Chinese population. AB - Angiotensin-converting enzyme has shown altered activity in patients with neurological diseases. An insertion/deletion (I/D) polymorphism of the DCP1 gene encoding angiotensin-converting enzyme has been reported to be associated with the risk for Alzheimer's disease (AD), but ambiguous results have also been presented. We conducted a case-control study in a sample composed of 192 sporadic AD patients and 195 age- and sex-matched controls from Chinese Han population in Beijing and Xi'an districts to investigate the possible effect of the polymorphism. Our data revealed no association between the DCP1 polymorphism and AD risk in the total sample. There was no significant difference in the DCP1 allele or genotype frequencies between cases and controls when stratified by gender and APOE epsilon4 status. However, the D allele and D/D genotype were more frequent among AD patients between 66 and 70 years compared with controls (D allele: OR=2.8, 95% CI=1.5-5.2, p=0.001; D/D genotype: OR=5.9, 95% CI=1.7-19.9, p=0.002). Our results provided new proof that the DCP1 D allele was a probable risk factor for late-onset AD. Its role was independent and was limited to the population at a certain age. PMID- 15832038 TI - Physical activity and postural control in the elderly: coupling between visual information and body sway. AB - BACKGROUND: Aging is characterized by a decline in the postural control performance, which is based on a coherent and stable coupling between sensory information and motor action. Therefore, changes in postural control in elderlies can be related to changes in this coupling. In addition, it has been observed that physical activity seems to improve postural control performance in elderlies. These improvements can be due to changes in the coupling between sensory information and motor action related to postural control. OBJECTIVE: The purpose of this study was to verify the coupling between visual information and body sway in active and sedentary elderlies. METHODS: Sixteen sedentary elderlies (SE), 16 active elderlies (AE) and 16 young adults (YA) were asked to stand upright inside a moving room in two experimental conditions: (1) discrete movement and (2) continuous movement of the room. RESULTS: In the continuous condition, the results showed that the coupling between the movement of the room and body sway was stronger and more stable for SE and AE compared to YA. In the discrete condition, SE showed larger body displacement compared to AE and YA. CONCLUSIONS: SE have more difficulty to discriminate and to integrate sensory information than AE and YA indicating that physical activity may improve sensory integration. PMID- 15832039 TI - Age of patients with papillary thyroid carcinoma: is it a key factor in the development of variants? AB - BACKGROUND: Although the majority of papillary thyroid carcinomas (PTC) have an excellent prognosis, certain aggressive variants do occur. The tall cell variant (TCV) of PTC, which tends to occur in elderly patients, is the most common among the aggressive variants and the most aggressive of all variants. OBJECTIVE: To compare the age of patients with major variants of PTC and correlate age with certain cytomorphological features of diagnostic importance. METHODS: 53 PTC cases with available information on age and diagnosed by fine needle aspiration cytology were categorized into various variants following a review of smears. There were 6 TCV (>or=30% tall cells) cases, 8 cases with a significant tall cell component (sig. TCC; 10-29% tall cells), 16 usual variants (UV), 17 follicular variants (FV), and 6 miscellaneous variants. These groups were compared with respect to age (mean+/-SD, median). Further, the age of all patients with PTC was correlated with cytological features such as tall cell count and count of cells with reddish cytoplasm, intranuclear cytoplasmic inclusion, and nuclear grooves. RESULTS: The median age of patients with TCV, PTC with a sig. TCC, UV, FV and miscellaneous variants was 47.5, 44.5, 31.5, 34.0 and 42.0 years, respectively. The age of PTC (TCV) cases (52.0+/-17.84 years) and of those with a sig. TCC (45.9+/-12.12 years) was significantly higher than the age of PTC (FV) cases (34.2+/-8.21 years) (p=0.004 and 0.027, respectively). The combined age of PTC (TCV) and PTC (sig. TCC) cases (48.5+/-14.54 years) was significantly higher than the age of PTC (UV) cases (p=0.038) as well as of PTC (FV) cases (p=0.005). Six (42.9%) of the patients with PTC (TCV and sig. TCC) were >or=50 years of age as opposed to 3 (9.1%) of PTC (UV and FV) cases (p=0.0134). The age of patients with PTC in general showed a significant correlation with tall cell count (p<0.01). CONCLUSION: The age of the patients with PTC seems to play a role in the morphological manifestations of the neoplasm, which is related to the prognostic outcome. PMID- 15832040 TI - Age-related alterations in the carbohydrate residue composition of the cell surface in the unexposed normal human epidermis. AB - BACKGROUND: The results of the very few histochemical studies that have been performed so far on the lectin-binding profile of normal human epidermis are mostly controversial; thus, the carbohydrate residue composition of the cell surface in the latter still remains in dispute and the possible alterations in the epidermal lectin-binding profile are unknown. OBJECTIVE: To investigate the influence of age on the carbohydrate residue composition of the cell surface in unexposed normal human epidermis by means of lectin histochemistry. METHODS: Biopsy specimens obtained from the sun-protected (unexposed) buttock skin, divided into 5 age groups of 18 subjects each, were fixed in buffered formalin (10%) and embedded in paraffin. 4-mum sections were processed for histochemistry using a panel of six biotinylated lectins. RESULTS: In the unexposed normal human epidermis the concentration and distribution of cell surface beta-D-galactose, D galactose-beta-(1,3 N-acetylo-D-galactosamine), beta(1,4 D-N-acetylo-D glucosamine) and alpha-D-N-acetylo-D-galactosamine were almost identical in all age groups, whereas those of alpha-D-mannose, alpha-D-glucose and alpha-L-fucose revealed significant age-related differences. CONCLUSION: These findings may be due to an age-related decline in synthesis and/or transport of monosaccharides from the cytoplasm to the surface of epidermal cells. Thus, the corresponding lectins concanavalin A and Ulex europaeus agglutinin-I can only be used in comparative histochemical studies of the carbohydrate residue composition of the cell surface in the normal and pathological epidermis of individuals of the same age, whereas Ricinus communis agglutinin-I, peanut agglutinin, wheat germ agglutinin and Dolichos biflorus agglutinin, whose binding to carbohydrates is not affected by aging can be used in histochemical studies of carbohydrate residue composition of the cell surface in the normal and pathological epidermis in human subjects of any age. PMID- 15832041 TI - Treadmill training effects in different age groups following middle cerebral artery occlusion in rats. AB - BACKGROUND: Despite the increased understanding of treadmill training on stroke patients, its effects on different age groups are not clearly known. The present study presents such effects through a model of cerebral ischemia on young and old groups of rats. OBJECTIVES: To investigate the effect of treadmill training on young and old rats after cerebral ischemia caused by middle cerebral artery occlusion (MCAO). METHODS: Forty old (22-24 months of age) and 32 young (3-4 months of age) rats underwent the MCAO procedure for 60 min. Rats that survived the procedure were randomly assigned to a 1- or a 2-week treadmill training group, or a time-matched control group (n=6-8 for each group). The infarct volume was compared between the treadmill training and the control groups for both the young and old rats at 1 or 2 weeks. RESULTS: After treadmill training for 1 week, the mean infarct volume was 7.26+/-0.49 and 9.51+/-0.84% for the young and old rats, respectively. The 1-week treadmill training effect was significant in the young rats (p=0.0207) but not in the old rats (p=0.0840). The mean infarct volume was 6.84+/-0.51 and 7.63+/-0.52% for the young and old rats, respectively, after the 2-week treadmill training. Both the young and old rat groups demonstrated a significant reduction in the infarct volume compared with that of the control group (p=0.021 for the young group and p=0.039 for the old group) after 2 weeks of treadmill training. CONCLUSION: The present findings clearly demonstrate the different training effects of locomotor activity in reducing ischemic infarction in young and old rats. The delayed reduction in ischemic infarction in old rats was notable and may be attributable to the slow response of angiogenic and neurogenic mechanisms in the old rats. PMID- 15832042 TI - Effects of a long-term dietary xylitol supplementation on collagen content and fluorescence of the skin in aged rats. AB - BACKGROUND: Dietary xylitol has been shown to increase the amounts of newly synthesized collagen, and to decrease fluorescence of the collagenase-soluble fraction in the skin of both healthy and diabetic rats. As in diabetic rats, a decreased rate of collagen synthesis and increased collagen fluorescence has also been detected in the skin of aged rats. We hypothesize that dietary xylitol supplementation may protect against these changes during aging. OBJECTIVE: The purpose of the present study was to investigate whether a long-term dietary supplementation can protect against the decrease in the amounts of newly synthesized collagen, and against the increase in fluorescence in the collagenase soluble fraction in the skin of aged rats. METHODS: Twenty-four male Sprague Dawley rats were used in the study. After weaning, the rats were divided into 2 groups of 12 animals. The rats in the control group were fed a basal RM1 diet, while the rats in the experimental group were fed the same diet supplemented with 10% xylitol. After 20 months, the rats were killed and pieces of skin from their dorsal areas were excised. The thickness of the samples was measured with a micrometer screw gauge. The collagen contents of rat skin were measured as hydroxyproline, and glycosylation as fluorescent intensity of collagen. Statistical significances of the differences between the groups were determined using the unpaired t test. RESULTS: No general side effects were detected in the rats during the experimental period. The skin of the xylitol-fed rats was a little thicker than that of the control rats. The hydroxyproline content of the acid-soluble fraction was significantly greater in the xylitol group as compared to the controls. However, there were no significant differences in the hydroxyproline content of the collagenase-soluble fraction between the groups. The fluorescence of the collagenase-soluble fraction was significantly smaller in the xylitol-fed aged rats than in the aged rats fed the basal diet. CONCLUSIONS: The results of this study indicate that xylitol caused an increase in the amount of newly synthesized collagen and a decrease in collagen fluorescence in the skin of aged rats. PMID- 15832043 TI - Hierachy of needs of geriatric patients. AB - BACKGROUND: The aim of the study is to contribute to the improvement of the quality of care of geriatric patients. In order to be able to improve the geriatric care we have to know clearly the needs of the elderly patients. OBJECTIVE: The hierarchy of needs of geriatric patients in clinical circumstances had to be assessed by a psychometric technique based on a motivational approach. METHODS: The process was based on the method of paired comparisons, and a duly composed questionnaire was administered to the geriatric patients who were proven to have consistent thinking and surpassed a pre-established threshold in the Mini Mental State Examination. The evaluation of the responds leads to a numerical derivation (on an interval scale) of the hierarchy of needs. RESULTS: The received hierarchy of needs proved to oppose the general hypothesis of Maslow's motivation theory. CONCLUSION: This may induce a necessary shift in the approach to the care of the elderly, and accentuate the self-actualization and esteem needs beside the overtly emphasized physiological care. PMID- 15832044 TI - Non-invasive (transcutaneous) monitoring of PCO2 (TcPCO2) in older adults. AB - BACKGROUND: Transcutaneous measurements of arterial blood gases (ABG) may decrease the need for repeated arterial puncture in older patients treated for acute cardiac or pulmonary disorders. However, age-related changes in skin perfusion, metabolism, or thickness may alter the validity of the technique. OBJECTIVE: To analyse the agreement between transcutaneous and arterial measurement of PaO2 and PaCO2 in older adults. DESIGN: Prospective descriptive study performed in the intermediate-care unit of a geriatric university hospital and a pulmonary rehabilitation centre. METHODS: 40 patients, aged 82.5+/-8 years (66-97), hemodynamically stable, without vasopressor treatment, underwent simultaneous measurement of arterial blood gases (ABG) and transcutaneous CO2 (TcPCO2) and O2 (TcPO2) with a Radiometer TINA TCM3 capnograph, and a probe T degrees set at 43 degrees C. RESULTS: Correlation between PaCO2 and TcPCO2 was high (r2=0.86) with a low bias (-0.1 mm Hg) and limits of agreement quite compatible with clinical use: (8.3; -8.5 mm Hg). The probe was well tolerated without any cutaneous lesion even after prolonged recordings (up to 8 h). Conversely, although TcPO2 and PaO2 were significantly correlated, the variability around the regression line precludes the use of transcutaneous measurements for monitoring PaO2)in a clinical setting. CONCLUSION: In older subjects, TcPCO2 (but not TcPO2) measurements are reliable when repeated assessment of ABG is warranted. PMID- 15832045 TI - Nutritional supplementation of very old people at hospital discharge increases muscle strength: a randomised controlled trial. AB - BACKGROUND: Undernutrition is common in older people admitted to hospital, but little is known about how nutritional state changes after discharge. OBJECTIVE: This randomised controlled trial was designed to examine the effect of oral nutritional supplementation of undernourished very old people prepared to take supplements after hospital discharge following acute illness. METHODS: Participants aged>or=75 years with a BMIor=5% during an acute hospital stay were allocated at random to either oral nutritional supplementation for 8 weeks from hospital discharge or to usual care. Primary outcome was change in weight, secondary outcomes were handgrip strength and anthropometry. RESULTS: Of 198 patients eligible to participate, 136 patients (mean age 85 years) were randomised and 76/136 (56%) completed the study. Twenty percent (13/66) of the intervention group withdrew after only 2 weeks, citing intolerance of the supplements. Using intention-to-treat analysis, body weight increased by a mean of 1.6 and 2.2% in the control and intervention groups, respectively, but this between-group difference was not significant (p = 0.188). However, handgrip strength increased more (p=0.055) in the intervention group (13.9%) than in the control group (7.2%). CONCLUSIONS: Oral nutritional supplementation was associated with a greater increase in handgrip strength than in non-supplemented controls and this observation merits further study. PMID- 15832046 TI - Modifiable factors for the length of life with disability before death: mortality retrospective study in Japan. AB - BACKGROUND: Past studies have measured and described the length of life with disability before death, but there has been no study of the relationship between modifiable lifestyle factors and duration of disability. OBJECTIVE: To examine whether there are modifiable factors influencing the length of life with disability before death. METHODS: The study was designed as a retrospective observation of the deceased who had earlier been enrolled in a prospective cohort study. During the follow-up period (1996-1999), we documented 781 deaths among those who were 70-79 years of age at the baseline survey in 1994 (n=10,216). In 2000, we interviewed family members of the deceased about the duration of the subjects' disability before death (n=655). RESULTS: The median duration of disability before death was approximately 6 months. Both higher Body Mass Index (BMI) and shorter time spent walking were significantly associated with an increased risk of long-term disability (more than 6 months). The odds ratios of long-term disability were 1.3 in those with BMI 20-25 and 2.1 in those with BMI>25, compared with BMI<20. The odds ratios of long-term disability were 1.3 in those walking for 0.5-0.9 h/day and 1.7 in those walking for <0.5 h/day, compared with those walking for >1.0 h/day. These relationships were unchanged after stratification for causes of death. CONCLUSION: Weight control and walking in later life may shorten the length of life with disability before death. PMID- 15832047 TI - Nutritional status and a 3-year follow-up in elderly receiving support at home. AB - BACKGROUND: Elderly receiving public services and care are often frail, suffer from chronic diseases, and sustain a high risk for malnutrition. OBJECTIVE: To evaluate nutritional status and long-term outcome in elderly living at home. METHODS: Of 507 eligible subjects receiving home care in five Swedish municipalities, we examined 353 (age 82+/-7 years, 64% females). The subjects were interviewed, and the nutritional status was assessed by means of the Mini Nutritional Assessment (0-30 points; the lower the score, the greater the risk). The Mini Nutritional Assessment consists of 18 questions concerning, e.g., anthropometry (body mass index or BMI; kg/m2) and global and dietary issues. The mortality was evaluated in 224 study participants after a 3-year period. In one municipality, 31 of 64 elderly were reexamined after 3 years. RESULTS: 8 and 41% of the elderly were assessed as malnourished or at risk of malnutrition, respectively. BMIs <20 and <23 were found in 12 and 31% of the subjects, respectively. Chewing and swallowing problems and reduced appetite were more often reported by those at risk of being malnourished compared with the well nourished study participants (p<0.001). Meals-on-wheels services were given to one third, of whom 66% used one portion for several meals. The 3-year mortality was 50% for those who were malnourished, 40% for those at risk of malnutrition, and 28% for the well-nourished group (p<0.05). The corresponding mortality was 50% for subjects with a BMI<20, 35% for those with BMIs 20-28, and 27% for those with a BMI>28 (p=0.05). After 3 years, a weight loss of 4.0+/-5.8 kg was registered (p<0.001). CONCLUSIONS: About half of the home-living elderly with public support were malnourished or were at risk of malnutrition. The malnourished subjects often had problems during mealtimes and seldom ate full meals. Elderly with a BMI>28 displayed the lowest risk of death within 3 years. PMID- 15832048 TI - Is it possible to predict one-year survival in centenarians? A neural network study. AB - BACKGROUND: Human life expectancy is constantly increasing: the challenge for modern geriatric medicine is to identify the means to reach successfully extreme longevity. OBJECTIVE: To determine which are the survival determinants in centenarians using a neural network. METHODS: Sample of 110 centenarians living in Rome, mean age 101.6 years (SD=1.8) with a sex ratio males:females of 1:3. We administered an extensive health interview (lasting 1-2 h) to each subject. The questionnaire, carried out according to the Geriatric Multidimensional Assessment, is made up of 100 items including a comprehensive health and psychosocial assessment aimed at various topics of general health and well-being and some scales used in geriatric practice. We applied several three-layered feed forward neural networks by mixing in different ways the most important of the 100 items. RESULTS: The most predicting powered net is the one constructed with 23 variables regarding comorbidity, cardiovascular risk factors, cognitive status, mood, functional status and social interactions, which therefore are strictly related to survival in centenarians. CONCLUSION: Survival in longevity is a complex biological phenomenon, which is an ideal field for using the neural network as a statistic method. The net shows us that the maintenance of social relationships even in presence of disability is of major importance for survival in the oldest old. PMID- 15832049 TI - Cross-domain variability of cognitive performance in very old nursing home residents and community dwellers: relationship to functional status. AB - BACKGROUND: Recent evidence suggests that cross-domain variability in cognition may be related to subsequent cognitive decline beyond mean performance levels in cognitive tasks. OBJECTIVES: To examine age-related changes in cross-domain variability across cognitive task performance in very old nursing home residents in contrast to community-dwelling older adults. To explore the relationship between cross-domain variability in cognition and functional disability in very old age. METHODS: 204 very old (82.00+/-8.51 years) residents from the Jewish Home and Hospital, Bronx, N.Y., and 376 community-dwelling older adults of similar age (86.75+/-5.84 years) were tested on a cognitive battery. Cross-domain variability scores were computed across the cognitive tasks. Functional disability measures were derived from the CDR (Clinical Dementia Rating) Scale. RESULTS: Whereas oldest old community-dwelling adults showed a decrease in variability across domains with age, variability increased with age in nursing home residents, irrespective of the level of cognitive performance. Cross-domain variability was associated with functional disability beyond the effects of age, gender, education, dementia status, residential status, and level of cognitive performance. CONCLUSION: Findings suggest that cross-domain variability in cognition is related to functional decline with age. Cross-domain variability in cognition may be a prominent predictor for the development of functional decline in very old adults. PMID- 15832050 TI - Celiac disease in the elderly. PMID- 15832051 TI - Lack of de novo sensitization to tropomyosin in a group of mite-allergic patients treated by house dust mite-specific immunotherapy. AB - BACKGROUND: There is some evidence that specific immunotherapy may induce de novo sensitization to proteins contained in the allergen extract. This raises the possibility that the novel sensitization to cross-reacting allergens may cause food allergies. OBJECTIVE: The aim of this study was to assess whether injection of mite-specific immunotherapy (SIT) may induce sensitization to shrimp tropomyosin. METHODS: Seventy mite-allergic subjects who did not show any shrimp sensitization at baseline were followed up for at least 3 years. Thirty-one of them underwent a 3-year SIT course, 39 served as controls. At the follow-up visit, SPTs with both fresh shrimp and commercial extract were performed, and patients underwent an open oral challenge with shrimp. RESULTS: Neither patient nor control had become sensitized to shrimp allergens at the end of the study period. Both patients and controls reported the regular intake of crustaceans and/or molluscs. CONCLUSION: Injection SIT with house dust mite extracts does not seem to induce de novo tropomyosin sensitization in mite-allergic patients. PMID- 15832052 TI - Analysis of growth factors and inflammatory cytokines in exhaled breath condensate from asthmatic children. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), AA isoform of platelet derived growth factor (PDGF-AA), and epidermal growth factor (EGF) are involved in the pathogenesis of airway inflammation in asthma. These molecules are closely associated with cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-4. This study investigates the relation between childhood asthma and levels of these mediators in exhaled breath condensate (EBC). METHODS: EBC was collected from asthmatic children and controls using a disposable collection kit, and the concentrations of VEGF, PDGF-AA, EGF, TNF-alpha and IL-4 in EBC were measured using sandwich enzyme immunoassays. Exhaled nitric oxide concentration was measured by a chemiluminescence analyzer. RESULTS: Thirty-five asthmatic patients aged between 7 and 18 years and 11 controls were recruited. Sixteen patients had intermittent asthma (IA) whereas 19 of them suffered from persistent asthma (PA). A significant correlation was found between IL-4 and TNF-alpha in EBC (rho = 0.374, p = 0.010). PDGF-AA levels in EBC were higher in subjects with diminished FEV1 (p = 0.023) whereas IL-4 concentrations were increased in asthmatics (p = 0.007) as well as subjects with increased plasma total IgE (p = 0.033). Patients with PA receiving high-dose inhaled corticosteroid (ICS) had higher EBC IL-4 concentration than those on low-dose ICS (p = 0.007). Linear regression revealed that PDGF-AA levels in EBC were negatively associated with FEV1 percentage (beta = -0.459, p = 0.006) among the asthmatic patients. CONCLUSIONS: IL-4 in EBC is increased in childhood asthma, and growth factors are detectable in a significant proportion of these children. Increased PDGF-AA is found in asthmatics with more severe airflow limitation. PMID- 15832053 TI - Epithelial defence by gamma delta T cells. AB - Gamma delta T cells constitute a separate lineage of T lymphocytes which differ from conventional alpha beta T cells with regard to T cell receptor (TCR) repertoire and tissue localization. In murine skin, gamma delta T cells expressing a canonical V gamma5 TCR are abundant and contribute as so-called dendritic epidermal T cells to local immune surveillance. In humans, major subsets of gammadelta T cells are recognized on the basis of their TCR V delta usage. While V delta2 cells dominate in the peripheral blood, V delta1 cells are preferentially localized in mucosal tissue including the intestinal epithelia. In this article we summarize basic features of intraepithelial gamma delta T cells and discuss their possible role in epithelial defence. PMID- 15832054 TI - Histamine in allergic inflammation and immune modulation. AB - Histamine, originally considered as a mediator of acute inflammatory and immediate hypersensitivity responses has also been demonstrated to affect chronic inflammation and regulate several essential events in the immune response. On the other hand, various cytokines control histamine synthesis, release and expression of histamine receptors (HRs). The cells involved in the regulation of immune response and hematopoiesis express HRs and also secrete histamine, which can selectively recruit the major effector cells into tissue sites and affect their maturation, activation, polarization and effector functions leading to chronic inflammation. Histamine, acting through its receptor type 2, positively interferes with the peripheral antigen tolerance induced by T regulatory cells in several pathways. Histamine also regulates antigen-specific Th1 and Th2 cells, as well as related antibody isotype responses. The diverse effects of histamine on immune regulation are due to differential expression and regulation of four HRs and their distinct intracellular signals. In addition, differences in affinities of these receptors are highly decisive on the biological effects of histamine and agents that target HRs. This article highlights the findings leading to a change of perspective in histamine immunobiology. PMID- 15832055 TI - Vascular wall remodeling in patients with supravalvular aortic stenosis and Williams Beuren syndrome. AB - Supravalvular aortic stenosis (SVAS) and Williams Beuren syndrome (WBS) can be considered as inherited diseases affecting the whole arterial tree and causing narrowing of the vessels. It has been reported that abnormal deposition of elastin in arterial walls of patients with SVAS and WBS leads to increased proliferation of arterial smooth muscle cells (SMC), which result in the formation of hyperplastic intimal lesions. In this work, we conducted morphological and morphometrical analysis with stenotic aortas from patients suffering from SVAS and WBS and from healthy control subjects and demonstrated that the amount of elastic fibers and the loss of integrity of vascular elastic fibers in the aortas reflect similar changes in the skin of patients with SVAS or WBS, as reported in our previous work conducted on skin in these pathological states. On the other hand, we conducted investigations on metalloproteinases (MMP2, MMP9, MMP7) and their specific tissue inhibitors TIMP1 and TIMP2 to verify their possible involvement in the etiopathogeny of SVAS and WBS. We particularly evidenced an altered MMP9/TIMP1 balance in favor of matrix degradation which could facilitate SMC migration and neointimal hyperplasia. Our findings suggest that elastinolytic enzymes secreted by arterial SMC, possibly including matrilysin 1, are critical for the development of arterial lesions in SVAS and WBS and contribute to perpetuate arterial stenosis in either SVAS or WBS. PMID- 15832056 TI - Intraluminal application of vascular endothelial growth factor enhances healing of microvascular anastomosis in a rat model. AB - BACKGROUND: Early reconstitution after injury to the endothelium is an important feature for reducing a number of vessel wall pathologies. We investigated the effect of vascular endothelial growth factor (VEGF) and its impact on the vascular remodeling process and reendothelialization after microsurgery. METHODS AND RESULTS: Microvascular anastomosis was performed in the rat femoral artery. One group was treated with intraluminal administration of VEGF and the other with vehicle. We investigated morphological, ultrastructural and immunohistochemical changes of the vascular wall and the reendothelialization process. After 10 days, reendothelialization was significantly faster in VEGF-treated rats. Transmission electron microscopy revealed a complete healing in contrast to vehicle-treated vessels. Moreover, extracellular matrix proteins, such as fibronectin, collagen types I, III and IV, were significantly increased. Furthermore, VEGF treatment significantly induced VEGF receptor 2, flk-1, osteopontin and TGF-beta(1) proteins. CONCLUSIONS: Our data clearly document for the first time that intraluminal treatment with VEGF is beneficial to the healing process in vascular microsurgery. Osteopontin and TGF-beta(1), both induced by VEGF, may play an important role in the vascular remodeling process. Our results provide clear evidence that VEGF application may represent a useful strategy in accelerating reendothelialization and improving vascular healing after microsurgery. PMID- 15832058 TI - Dementia, stroke and Parkinson's disease in Spanish populations: a review of door to-door prevalence surveys. AB - We identified 14 door-to-door prevalence surveys on dementia, parkinsonism or stroke in Spanish populations fulfilling specific criteria and combined selected age- and sex-specific data using logistic regression and taking Pamplona as a reference. The prevalence of dementia and of Alzheimer's disease varied significantly with space. However, the largest variation was seen for vascular dementia: odds ratio (OR) and 95% confidence interval (CI) for Gerona were 6.42 (3.23-12.3) in women and 2.30 (1.10-4.79) in men. Stroke was particularly frequent among Arevalo's women, with OR 2.10 and 95% CI 1.26-3.49. The prevalence of Parkinson's disease was twofold higher in Cantalejo. Although differences in methodology make the interpretation of results problematic, the prevalence of stroke and vascular dementia in Spain seems to vary spatially, indicating a space for prevention. PMID- 15832059 TI - Mortality from human transmissible spongiform encephalopathies: a record linkage study. AB - To evaluate the ability of the Italian Creutzfeldt-Jakob (CJD) register to detect human transmissible spongiform encephalopathy (TSE) cases we compared mortality data from the CJD register with those obtained from death certificates collected by the Italian National Census Bureau (ISTAT) between 1993 and 1999. We used the method of record linkage to compare and integrate data from these two sources. The integrated estimate of TSE deaths was 457: 183 deaths recorded by the CJD register and ISTAT, 210 cases only by the CJD register, and 64 cases only by ISTAT. The average integrated estimated mortality rate was 1.58 deaths per million people per year over the study period and peaked in 1999 at 2.13. This figure is similar to that obtained from data from the CJD register alone in the years 2000-2002. The increase in mortality rates is likely due to an improvement in case ascertainment. The misclassification of cases by ISTAT was above 50% from 1996 onward, suggesting that using only death certificates is not a reliable way to monitor TSE cases in Italy. PMID- 15832060 TI - Morphometric analysis of gray matter volume in demented older adults: exploratory analysis of the cardiovascular health study brain MRI database. AB - We tested the feasibility of a fully automated brain MRI voxel count technique- automated labeling pathway (ALP)--in a sample of 15 demented and 13 cognitively normal women (age 75-85 years) participating to the Cardiovascular Health Study (CHS). We hypothesized that ALP would replicate well-established findings of the anatomical correlates of dementia. In particular, we hypothesized that ALP volumetric measures would: (1) significantly differ between cognitively normal and demented women in those brain areas that are established markers for diagnosis of dementia (temporal and medial temporal lobes, hippocampus, amygdala and parahippocampus) but not in other brain areas (e.g., occipital lobe, visual cortex, motor cortex) and (2) correlate with visual ratings of brain disease which have been previously collected as part of the CHS. ALP required minimal operator intervention (input of brain images and verification of misalignments) and employed computer time of about 1 h per brain. ALP detected significant focal volumetric differences in the limbic system (p values between groups for hippocampus and parahippocampus: 0.002 and 0.005, respectively), temporal lobe (p < 0.0001) and caudate (p = 0.009), but not in other brain areas (e.g. occipital lobe, visual or motor cortex). Furthermore, ALP measures of medial temporal lobe atrophy strongly correlated with CHS visual ratings of ventricular enlargement (r(2) = 0.6, p = 0.002 for medial temporal lobe). In conclusion, ALP-detected focal brain atrophy was strongly associated with dementia. Because of its fully automated design, ALP technique is an ideal candidate to assess whether volumetric measures of specific areas can discriminate dementia better than currently available measures of global brain atrophy in large epidemiological studies. PMID- 15832061 TI - Cerebral ischemic events in patients with advanced lung or prostate cancer. AB - Conventional stroke risks are thought responsible for most cerebral ischemic events (CIE) in adult cancer patients. Also suspected as a risk is cisplatin chemotherapy, alone or in combination with tumor angiogenesis inhibitor. We investigated whether treatment or tumor characteristics, independently of conventional stroke risks, are associated with CIE in a retrospective cohort study of 1,559 patients with advanced non-small cell lung cancer or hormone refractory prostate cancer followed during 3 clinical trials of matrix metalloprotease inhibitor (prinomastat) versus placebo, with chemotherapy (gemcitabine/cisplatin, paclitaxel/carboplatin or mitoxantrone/prednisone). During 11,907 patient-months, 28 CIE (17 cerebral infarction, 11 transient ischemic attack) were diagnosed in 24 patients, all but 1 over 55 years. Neither prinomastat, platinum-based chemotherapy nor their combination was associated with CIE after age 55. However, such events were predicted by the presence of distant metastases in the liver or lungs and not in distant lymph nodes (hazard estimate 4.6, 95% CI 2.0-10.5, adjusted for conventional stroke risks). Further studies are needed to verify this preliminary finding and determine its generalizability to advanced tumors other than lung or prostate cancer. PMID- 15832062 TI - Vascular endothelial growth factor expression, vascularization and proliferation in paragangliomas. AB - OBJECTIVE/HYPOTHESIS: Paragangliomas are heavily vascularized tumors, and the expression of VEGF (vascular endothelial growth factor) has been reported. The aim of our study was to extend the available database of VEGF expression in paraganglioma, to add correlated data concerning vessel density and proliferative activity, and to draw conclusions concerning the mechanisms resulting in tumor vascularization and growth. STUDY DESIGN: Semiquantitative histopathologic examination of paraganglioma specimens obtained from surgical cases. METHODS: Paraffin-embedded paragangliomas were analyzed by immunohistochemistry. Fourteen consecutive samples were hybridized with VEGF-, CD31- and Ki67-specific antibodies, and visualized by diaminobenzidine staining. Vessel density was determined by counting CD31-positive vessels and proliferation by quantification of Ki67-positive cells. RESULTS: Ten out of 14 samples were positive for VEGF. In this group, vessel density was up to 5 times as high and proliferative activity was about twice as high as in the VEGF-negative group. CONCLUSIONS: We observed higher CD31 and Ki67 counts in VEGF-positive tumors, but statistical significance could not be assessed due to low sample numbers. These data might suggest a contribution of VEGF secreted by paragangliomas to tumor vascularization and possibly proliferation. The clinical impact of VEGF expression analysis has to be proven in future studies. PMID- 15832063 TI - The effectiveness of cognitive therapy for schizophrenia: what can we learn from the meta-analyses? PMID- 15832064 TI - Cognitive behaviour therapy for schizophrenia -- a review of development, evidence and implementation. AB - BACKGROUND: Schizophrenia and other psychotic disorders were once thought to be impervious to psychological treatments; however, there is accumulating evidence that cognitive behaviour therapy (CBT) can result in significant clinical benefit to these patients. AIM: This paper aims to describe the development and adaptation of CBT in the treatment of schizophrenia, to summarise the evidence to support CBT as a viable treatment and to outline some of the issues in 'rolling out' this treatment into normal clinical settings. RESULTS: A number of clinical models of CBT have been developed and these typically consist of a variety of clinical methods with different models providing differing emphasis. Twenty controlled trials of CBT in schizophrenia in which 739 patients were included are reviewed. These studies have a mean effect size for CBT of 0.37 (SD 0.39). There is consistent evidence that CBT reduces persistent positive symptoms in chronic patients and may have modest effects in speeding recovery in acutely ill patients. The evidence of CBT reducing relapse rates is equivocal, although targeted early intervention is promising. The available evidence suggests that CBT can be utilised effectively in routine clinical practice. However, the dissemination of novel psychological treatments into widespread clinical practice is not without difficulty, and issues pertaining to the 'roll-out' of CBT are discussed. CONCLUSIONS: CBT as an adjunct treatment shows considerable promise for the future treatment of schizophrenia. PMID- 15832065 TI - Rethinking models of psychotropic drug action. AB - Theoretical assumptions about how psychotropic drugs 'work' are rarely discussed explicitly. In a 'disease-centred model,' drugs are believed to work by acting on a disease process. In contrast, in a 'drug-centred model,' the characteristic physiological, behavioural and subjective effects of drugs are used to define drug action. The therapeutic value of a drug stems from the usefulness of these effects in clinical situations. The disease-centred model appears dominant but has weaknesses: (1) it cannot logically justify the use of drugs since major pathophysiological hypotheses were derived from selectively observed actions of drugs; (2) comparisons between drugs believed to have specific effects in certain conditions and drugs thought to have non-specific effects fail to support it; (3) outcome measures for various disorders include items responsive to non-specific drug effects; (4) studies with healthy volunteers describe characteristic drug induced states independently of a psychiatric diagnosis; (5) animal tests show effects with agents not usually thought of as specific treatments for the conditions modelled by tests. This article offers suggestions to develop a drug centred model and discusses its potential impact on clinical practice. PMID- 15832066 TI - Internet-guided self-help with or without exposure therapy for phobic and panic disorders. AB - BACKGROUND: As many sufferers from phobic and panic (phobia/panic) disorders cannot get to suitable therapists, routine aspects of therapy were delegated to internet-accessed computer-aided self-help with or without exposure instructions. METHODS: Phobia/panic referrals were randomised to computer-aided self-help via the internet at home in a 2:1 ratio either by self-exposure cognitive behaviour therapy (CBT) [FearFighter (FF), n = 45] or by minimal CBT without exposure [Managing Anxiety (MA), n = 23]. All had brief backup phone advice from a clinician concerning their computer guidance. RESULTS: On self-ratings and blinded assessor ratings, patients improved equally with each form of self-help over 10 treatment weeks but significantly more on 5 out of 10 measures by week 14 (1-month follow-up) when the self-help included self-exposure instructions than when it did not. In accord with this, standardised effect sizes (Cohen's d) indicated superiority of FF over MA on 5 measures by week 14. Satisfaction with treatment in all patients pooled correlated positively with improvement after treatment and at 1-month follow-up. CONCLUSIONS: At the end of treatment, computer-aided CBT self-help at home via the internet plus brief live helpline support was effective with or without exposure instructions, and at 1-month follow-up it was more effective on some measures if exposure instructions had been included. Analysis is needed of how non-exposure CBT produced its shorter term effect. PMID- 15832067 TI - Effect of self-hypnosis on hay fever symptoms - a randomised controlled intervention study. AB - BACKGROUND: Many people suffer from hay fever symptoms. Hypnosis has proved to be a useful adjunct in the treatment of conditions where allergic phenomena have an important role. METHODS: Randomised parallel group study over an observation period of two consecutive pollen seasons. Outcome data include nasal flow under hypnosis, pollinosis symptoms from diaries and retrospective assessments, restrictions in well-being and use of anti-allergic medication. We investigated 79 patients with a mean age of 34 years (range 19-54 years; 41 males), with moderate to severe allergic rhinitis to grass or birch pollen of at least 2 years duration and mild allergic asthma. The intervention consisted of teaching self hypnosis during a mean of 2.4 sessions (SD 1.7; range 2-5 sessions) and continuation of standard anti-allergic pharmacological treatment. RESULTS: Of 79 randomised patients, 66 completed one, and 52 completed two seasons. Retrospective VAS scores yielded significant improvements in year 1 in patients who had learned self-hypnosis: pollinosis symptoms -29.2 (VAS score, range 0-100; SD 25.4; p < 0.001), restriction of well-being -26.2 (VAS score, range 0-100; SD 28.7; p < 0.001. In year 2, the control group improved significantly having learned self-hypnosis as well: pollinosis symptoms -24.8 (SD 29.1; p < 0.001), restriction of well-being -23.7 (SD 30.0; p < 0.001). Daily self-reports of subjects who learnt self-hypnosis do not show a significant improvement. The hazard ratio of reaching a critical flow of 70% in nasal provocation tests was 0.333 (95% CI 0.157-0.741) after having learnt and applied self-hypnosis. PMID- 15832068 TI - Tridimensional personality questionnaire factors in major depressive disorder: relationship to anxiety disorder comorbidity and age of onset. AB - OBJECTIVE: We used the Tridimensional Personality Questionnaire (TPQ) to study the relationship between temperamental traits and comorbid anxiety disorders as well as age of onset of major depressive disorder (MDD) in 263 patients with MDD. METHODS: Patients recruited for a large clinical study on MDD underwent a Structured Clinical Interview for DSM-III-R assessment and were administered the self-rated TPQ [mean age = 39.5 +/- 10.5 years, women = 138 (53%), initial 17 item Hamilton Rating Scale for Depression (HAM-D-17) score = 19.6 +/- 3.4]. The TPQ was scored for three previously identified factors -- harm avoidance (HA), novelty seeking (NS), and reward dependence (RD). Multiple linear regression methods were used to evaluate the relationship between TPQ factors and each comorbid anxiety disorder as well as between early-- vs. late-onset MDD, after controlling for age, gender and initial HAM-D-17 score (when these were related to the dependent variable in simple regressions). RESULTS: Social anxiety disorder in MDD was strongly associated with higher scores on HA and lower scores on NS and RD (t = 5.4, p < 0.0001; t = 2.6, p = 0.009; t = 2.2, p = 0.028, respectively). A diagnosis of generalized anxiety disorder in MDD was significantly related to higher HA scores (t = 2.8, p = 0.006). The presence of comorbid obsessive-compulsive disorder was associated with lower NS scores (t = 2.3, p = 0.023) as was that of comorbid panic disorder (t = 2.0, p = 0.051). Finally, the presence of simple phobias was associated with lower scores on RD (t = 2.4, p = 0.016). HA scores were higher in patients with early onset of MDD (adjusted p = 0.05). Early versus late onset of MDD was not significantly related to NS or RD scores. LIMITATIONS: Since our sample consisted of moderately depressed outpatients, our ability to generalize our findings to other populations is limited. CONCLUSIONS: Features of temperament are related to patterns of anxiety disorder comorbidity and age of onset among patients with MDD. Higher levels of HA and lower levels of RD and NS were associated with an increased risk of anxiety disorder comorbidity in our sample. HA may also be related to early onset of depression. PMID- 15832069 TI - Stressful life events, depression and demoralization as risk factors for acute coronary heart disease. AB - BACKGROUND: While the effect of psychological stress and depression on the course of heart disease is commonly recognized, the relationship between recent life events, major depression, depressive symptomatology and the onset of acute coronary heart disease (CHD) has been less considered. The aim of this study was to investigate the presence of stressful life events, major and minor depression, recurrent depression and demoralization in the year preceding the occurrence of a first acute myocardial infarction (AMI) and/or a first episode of instable angina and to compare stressful life events, also related with mood disorders, in patients and healthy controls. METHODS: 97 consecutive patients with a first episode of CHD (91 with AMI and 6 with instable angina) and 97 healthy subjects matched for sociodemographic variables were included. All patients were interviewed with Paykel's Interview for Recent Life Events, a semistructured interview for determining the psychiatric diagnosis of mood disorders (DSM-IV), a semistructured interview for demoralization (DCPR). Patients were assessed while on remission from the acute phase. The time period considered was the year preceding the first episode of CHD and the year before the interview for controls. RESULTS: Patients with acute CHD reported significantly more life events than control subjects (p < 0.001). All categories of events (except entrance events) were significantly more frequent. 30% of patients were identified as suffering from a major depressive disorder; 9% of patients were suffering from minor depression, 20% from demoralization. Even though there was an overlap between major depression and demoralization (12%), 17% of patients with major depression were not classified as demoralized and 7% of patients with demoralization did not satisfy the criteria for major depression. Independently of mood disorders, patients had a higher (p < 0.001) mean number of life events than controls. With regard to life events, the same significant difference (p < 0.001) compared to controls applied to patients with and without mood disorders. CONCLUSIONS: Our findings emphasize, by means of reliable methodology, the relationship between life events and AMI. These data, together with those regarding traditional cardiac risk factors, may have clinical and prognostic implications to be verified in longitudinal studies. PMID- 15832070 TI - Trauma work, sense of coherence, and positive and negative changes in therapists. AB - BACKGROUND: The negative effects of vicarious traumatization through trauma work are well known. However, a growing body of evidence testifies to the positive effects that have been reported following trauma. This study explored the potential positive effects of trauma work on therapists, with a focus on the associations of the personality construct 'sense of coherence' with both positive and negative changes. METHODS: The study examined the positive and negative effects of trauma work in an international sample of 85 therapists. Participants completed measures of their trauma work experience, sense of coherence and positive and negative changes they experienced as a result of their trauma work. RESULTS: A greater sense of coherence was associated with fewer negative changes and more positive changes. CONCLUSIONS: Adversarial growth in therapists deserves further enquiry to allow a more detailed understanding of the factors and processes involved. PMID- 15832071 TI - Worrying patterns of out-patient psychotropic drug prescribing in children and adolescents. PMID- 15832072 TI - Prevalence of bipolar disorders. PMID- 15832074 TI - Jan Mikulicz-Radecki: one of the creators of world surgery. AB - This paper presents the life and achievements of Professor Jan Mikulicz-Radecki and his contribution to European and world surgery. He was born in 1850. Four periods can be distinguished in his surgical career: (1) Vienna period, 1875-82, when he worked by the side of the great Theodor Billroth: he introduced a number of new diagnostic and operative techniques, aseptic and antiseptic procedures, published research papers. He made the first endoscope for examining the esophagus and stomach. (2) Cracow period, 1882-87, when he was head of the Department of Surgery: he inaugurated his work with a lecture in Polish, which started: Gentlemen, I have been accused that I do not know the Polish language- which is my mother tongue to me as well as to any of you. He published papers on the use of iodophorm for healing wounds, was the first surgeon who sutured a perforated gastric ulcer (1885) and invented pyloroplasty (1887), nowadays called Heinecke-Mikulicz pyloroplasty. (3) Konigsberg period, 1887-90: he improved the technique of gastric resection, worked on surgery for peptic ulcer and advocated aseptic and antiseptic procedures. (4) Wroclaw (Breslau) period, 1890- 1905: Mikulicz was appointed head of a newly founded Surgical Department. He rebuilt it and designed one of the largest and most modern operating theaters in Europe that time. He introduced silk gloves for operations for the first time in the world. In 1892 he described bilateral swelling of salivary and lacrimal glands, called Mikulicz's syndrome; in 1902 he published his experience with two-stage colon resection; and in 1904 he described ostitis fibrosa cystica juvenilis. Called "a king in the kingdom of surgeons", he was one of the fathers of the world's modern surgery; having a brilliant surgical mind, he was a pioneer of many new operations and an inventor of surgical tools, permanently assimilated in the world's surgery. PMID- 15832075 TI - Olfactory ensheathing cells and spinal cord repair. AB - The olfactory ensheathing cell is a specialized glial cell that assists in growth of the axons of the olfactory sensory neurons as they are generated and regenerated throughout adult life. There is increasing evidence in animal models that transplantation of olfactory ensheathing cell promotes recovery after transplantation into the injured spinal cord. Olfactory ensheathing cell transplants have promoted regrowth of axons across the injury site and led to recovery of functional behaviours including climbing, walking, reaching, and breathing. Most evidence comes from olfactory ensheathing cells derived from the olfactory bulb. This is an impractical site for human biopsy compared to the easy accessibility of olfactory ensheathing cells from the olfactory mucosa in the nose. Our experiments demonstrated that nasal olfactory ensheathing cells led to functional improvement after complete spinal cord transaction in rat. After devising methods to grow human olfactory ensheathing cells from nasal biopsy we recently initiated a Phase I clinical trial of transplantation into the human paraplegic spinal cord. PMID- 15832076 TI - New genetic causes of pheochromocytoma: current concepts and the clinical relevance. AB - Pheochromocytoma and paraganglioma are tumors of the autonomous nervous system mainly occurring in the adrenal medulla, but also in the extraadrenal paraganglias of the abdomen, thorax, neck and skull basis. The etiology comprises germline mutations of now 6 genes. About 10 years known are the RET gene susceptible for multiple endocrine neoplasia type 2, the VHL gene for von Hippel Lindau Disease, and the NF 1 gene for neurofibromatosis Recklinghausen (neuro- fibromatosis type 1). Since 2000 the genes for succinatedehydrogenase subunits SDHB, SDHC, and SDHD have been identified for paraganglioma syndromes type 4, type 3, and type 1 respectively. Investigations of series of pheochromocytoma patients identified germline mutations in one of the genes SDHB, SDHD, VHL and RET in 24% to 50% of the patients. Multifocal tumors, young age and positive family history, known features associated with inheritence, have not been present in all patients. Therefore, analyses of blood DNA for mutations in these genes are recommended. Positive tests provide the patients and their relatives with essential platforms for clinical care. Experiences in this field of medicine have shown that optimal management of patients with pheochromocytoma-associated syndromes is a high challenge. National registries may be instrumental in order to provide with adequate facilities. PMID- 15832077 TI - Trends in North American medical education. AB - Medical education in the United States of America (USA), and worldwide, is increasingly concentrating on the process and outcome of the educational experience. The first efforts to substantially improve medical education in the USA resulted in the Flexner Report in the early 1900s. The release of this report led to significant advancements in the quality of curriculum content. However, in the past few decades there has been increasing realization that delivery of content will not, by itself, assure the development of excellent physicians. As a result, there has been an increasing emphasis on the process, and, most recently, the outcome, of medical education. Process movements have examined the context and methods for teaching and learning. The problem-based learning movement is perhaps the most widely-known example of process trends. The latest trends in USA medical education focus on the outcomes of the learning process. At the forefront of this movement is the American Council on Graduate Medical Education (ACGME), which accredits all USA post-graduate training programs. Recently, the ACGME has defined a set of six core clinical competencies that all graduates must demonstrate. A second emerging trend is inter-professional education. Increasingly, healthcare is provided by inter-professional healthcare team, and students must be competent to function effectively in this setting. Many academic health centers are developing joint curricula to address this need. Medical education has evolved from a primary focus on content to an emphasis on process of teaching and learning, and will increasingly concentrate on educational outcomes. PMID- 15832078 TI - Establishment of an immortalized porcine granulosa cell line (PGV) and the study on the potential mechanisms of PGV cell proliferation. AB - In order to establish an immortalized granulosa cell line and to investigate the potential mechanisms of immortalized cell proliferation, simian virus (SV) 40 was used to infect porcine granulosa cells from small follicles (1-2 mm in diameter), and one colony was selected after four weeks of culture. The colony was digested with trypsin and the cells were cultured for more than 300 days (named PGV). The SV40 large T antigen gene and its products were confirmed in immortalized cells by Southern blotting and immunohistochemistry. Progesterone production was not detected in the conditioned culture media with follicle-stimulating hormone (FSH) and forskolin, possibly due to the lack of P450scc gene transcription as examined by Northern blotting. PGV cells responded significantly to the stimulation of sera (fetal bovine and horse sera) and protein kinase C (PKC) stimulators (PMA and OAG), while PKC inhibitors (staurosporine and calphostin C) blocked both sera and PKC stimulation. Phospholipase C (PLC) and phosphatidic acid phosphatase (PAP) inhibitors (U73122 and propranolol) significantly reduced PGV cell proliferation, while PMA restored PLC and PAP inhibition. These data suggest that diacylglycerol (DAG) is produced in PGV cells by PLD as well as by PLC, and that DAG then activates PKC stimulating the PGV cell cycle through yet unknown mechanisms. Thus, an immortalized granulosa cell line is very useful to study granulosa cells in vitro, as the cells are homogeneous and are a functionally defined population. PMID- 15832079 TI - Primary synovial sarcomas of the mediastinum: a clinicopathologic, immunohistochemical, and ultrastructural study of 15 cases. AB - A series of 15 cases of primary mediastinal neoplasms displaying histopathologic, immunohistochemical, and ultrastructural features of synovial sarcoma is presented. The patients' ages ranged from 3 to 83 years, with a male-to-female ratio of 2:1. Nine cases presented as anterior mediastinal masses with chest pain, shortness of breath, and pleural effusion, and 6 cases were in paravertebral location in the posterior mediastinum and presented with neck or back pain and pleural effusion. The tumors measured from 5 to 20 cm in greatest diameter and showed a tan white, soft to rubbery cut surface with areas of hemorrhage and necrosis and foci of gelatinous material. Four cases showed areas of cystic degeneration. In 7 cases, the tumors were well circumscribed; in 6 cases, the tumors grossly invaded the pleura, pericardium, heart, great vessels, chest wall, rib, and vertebra. Histologically, 5 cases displayed a biphasic growth pattern, with well-formed glandular elements admixed with a monotonous spindle cell population. Ten cases were exclusively composed of a monotonous atypical spindle cell proliferation. Immunohistochemical studies showed focal positivity of the tumor cells for cytokeratin and/or epithelial membrane antigen, and strong positivity for vimentin and bcl-2 in the spindle cells in all cases studied (10 of 10). Eight cases also showed focal positivity for CD99. Electron microscopic examination in 5 cases showed oval to spindle tumor cells with closely apposed cell membranes, abundant cytoplasmic intermediate filaments and rough endoplasmic reticulum, and immature desmosome-type cell junctions. Ten patients were treated by complete surgical excision and two by partial excision followed by radiation therapy. In 4 patients, the tumors were inoperable and treated with radiation therapy only. Clinical follow-up was available in 5 patients and showed local recurrence with metastases to lung, lymph nodes, and epidural space from 1 to 3 years in 4 cases and liver metastases and death due to tumor after 6 month in 1 case. Synovial sarcoma should be considered in the differential diagnosis of biphasic and monophasic spindle cell neoplasms of the mediastinum. PMID- 15832080 TI - Immunohistochemical stains for p63 and alpha-methylacyl-CoA racemase, versus a cocktail comprising both, in the diagnosis of prostatic carcinoma: a comparison of the immunohistochemical staining of 430 foci in radical prostatectomy and needle biopsy tissues. AB - The diagnosis of prostatic carcinoma and especially minimal prostatic carcinoma can sometimes be challenging on needle core biopsy and occasionally immunohistochemistry is an aid in the diagnosis. Immunostains, such as those directed against the basal cell marker p63 and, more recently, employing antibodies reactive with alpha-methylacyl-CoA racemase (AMACR), can be useful in this situation. The aim of this investigation was to assess the diagnostic utility of a p63/AMACR antibody cocktail and compare the staining pattern it produces with that using the individual antibodies alone. A retrospective review of 31 consecutive radical prostatectomy specimens and 150 prostate needle biopsy samples was performed to select histologic sections showing foci of prostatic carcinoma and/or minimal prostatic carcinoma, high-grade prostatic intraepithelial neoplasia (HGPIN), as well as common benign mimickers of prostatic carcinoma, to include atrophy and basal cell hyperplasia, especially with prominent nucleoli. Serial histologic sections from the corresponding paraffin blocks were stained with hematoxylin and eosin and by immunostains for p63, AMACR, and a prediluted antibody cocktail comprising both. The diagnostic utility of the cocktail was assessed, and the staining characteristics it produced were compared with those using the individual immunostains. In 430 foci, the cocktail produced a p63 staining profile identical to that using the single p63 antibody. Distinction of the nuclear p63 signal from the cytoplasmic AMACR localization was readily accomplished. There was an excellent agreement (kappa=0.91; P<0.0001) between the AMACR staining profile using the cocktail and the single AMACR antibody alone. The cocktail was very useful in highlighting prostatic carcinoma associated with HGPIN, flat and cribriform HGPIN, and distorted foci of minimal prostatic carcinoma. These data indicate that use of a p63/AMACR cocktail is essentially equivalent to use of each antibody separately for immunohistochemical confirmation of a diagnosis of prostatic carcinoma in needle biopsy. This cocktail would be of diagnostic utility when only limited tissue is available for immunohistochemical evaluation of small, diagnostically difficult foci in prostate needle biopsy tissue. PMID- 15832081 TI - High-grade neuroendocrine carcinoma of the ampulla of vater: a clinicopathologic and immunohistochemical analysis of 14 cases. AB - We describe the clinical and pathologic features of 14 cases of high-grade neuroendocrine carcinoma (HGNEC) of the ampulla of Vater classified according to WHO classification of lung tumors into small cell carcinoma (SCC, 6 cases) and large cell neuroendocrine carcinoma (LCNEC, 8 cases) types. The immunohistochemical findings were compared with those of 13 cases of primary poorly differentiated ampullary adenocarcinomas (PDACA) lacking neuroendocrine morphology. The mean age of 10 males and 4 females was 70 years. The mean tumor size was 2.5 cm. Ten of 13 patients had lymph node metastases (mean, 2.3 nodes involved). Documented sites of distant metastases included brain and liver. Overall, 64% of patients with ampullary HGNEC died of disease (mean follow-up, 14.5 months). Four patients had no evidence of disease after resection (mean, 20 months). Half of the tumors were associated with adenomas of the adjacent mucosa, 2 with high-grade dysplasia. Two HGNECs were combined with a conventional adenocarcinoma and another with a squamous cell carcinoma component. By immunohistochemistry, the HGNECs were positive for cytokeratins (AE1/AE3, 100%; Cam5.2, 67%; CK7, 87%; CK20, 38%), similar to the pattern found in PDACAs. p27 expression was lost in 1 case of HGNEC and in all PDACAs. Retinoblastoma (Rb) protein expression was lost in 60% of HGNECs and in none of the PDACA cases. In conclusion, HGNECs of the ampulla are rare (2%-3% of ampullary tumors in our material). The clinical course parallels that of their pulmonary counterparts and appears to be worse than that of locally advanced ampullary adenocarcinomas. The association with adenoma and or conventional adenocarcinoma components may suggest a common pathway in the initial carcinogenesis of these two types of tumors. Loss of Rb expression, a characteristic finding in pulmonary SCCs, is present in almost half of ampullary HGNECs. In contrast, p27 expression is lost in PDACAs and retained in most HGNECs. Thus, there are differences in the molecular phenotypes of these two types of ampullary carcinoma, supporting the distinction of poorly differentiated carcinomas with a neuroendocrine phenotype from those without. PMID- 15832082 TI - Epstein-barr virus infection of monocytoid B-cell proliferates: an early feature of primary viral infection? AB - Monocytoid cells are a subset of B lymphocytes with characteristic morphology and immunophenotype, which proliferate in a broad variety of reactive lymph node conditions. Up to now, no direct infection of these cells by a pathogenic organism has been identified. We found in a series of lymph node specimens from 13 patients having in common a prominent monocytoid B (MCB)-cell reaction in the absence of epithelioid cells or necrosis evidence that Epstein-Barr virus (EBV) can infect MCB cells. Clinical and serologic findings indicative for an acute EBV infection were reported only in 2 cases. By in situ hybridization and immunohistology, a number of MCB cells in all cases was found to contain EBV encoded small nuclear RNAs and to express the EBV-encoded nuclear antigen (EBNA) 2 and, in lesser extent, the latent membrane protein-1. EBV-transcripts and proteins were also detectable in a number of extrafollicular B blasts and germinal center B cells in 10 of 13 cases. Our findings imply that an EBV infection of MCB cells associated with predominant EBNA-2 expression represents an early sign of primary infection and that it should be included in the differential diagnosis of activated lymph nodes with prominent MCB-cell reaction in the absence of epithelioid cells. PMID- 15832083 TI - Impact of colloid response on survival after preoperative radiotherapy in locally advanced rectal carcinoma. AB - Neoadjuvant therapy for rectal carcinoma modifies morphology and natural history of the tumor. Colloid response defined by predominant colloid changes with or without residual tumor cells is a form of tumor response whose impact on survival is unknown. This study evaluated influence of tumor histologic response, especially of colloid response, on survival in patients treated by long-course preoperative radiotherapy for rectal cancer. In 200 patients with uT3-T4 or N1 rectal carcinomas, influence of type of surgery, dose of radiotherapy, residual tumor size, surface tumor aspect, tumor response (downstaging vs. colloid or no response), tumor grade, vascular and neural invasion, circumferential margin, and postoperative chemotherapy on 5-year overall and disease-free survival were studied by univariate and multivariate analyses. A colloid response was observed in 20% of the cases. Tumor response, circumferential margin, and vascular invasion were independently associated with the disease-free survival. Patients with downstaging had a better disease-free survival than patients without response (80% vs. 54%), whereas those with colloid response had an intermediate survival (64%). After colloid response, the rate of recurrence was similar to patients with downstaging for local recurrence (0%-3%) and to those with no response for distant recurrence (28%). After preoperative radiotherapy for rectal cancer, survival and type of recurrence are influenced by the tumor response. The intermediate natural history of patients with colloid response suggests taking colloid response into account in postoperative tumor staging to optimize adjuvant therapy. PMID- 15832084 TI - Intraductal tubular adenoma of the pancreas, pyloric gland type: a clinicopathologic and immunohistochemical study of 6 cases. AB - The intraductal tubular adenoma (ITA), pyloric gland type, of the pancreas is an uncommon benign tumor, akin to the pyloric gland type adenoma of the gallbladder. We report 6 cases of ITA of the pancreas: 3 male and 3 female aged 50 to 79 years (mean, 63.5 years; median, 65 years); all were examined clinicopathologically. Four patients showed no symptoms, but appetite loss and/or general fatigue presented in two. Grossly, all tumors formed a localized polypoid mass protruding into the lumen of the dilated pancreatic duct. Five of the six tumors were found within the main duct, and the other arose within the branch duct of the pancreas. Microscopically, the tumors were composed of closely packed tubular glands resembling pyloric type glands. They were lined by columnar or cuboidal epithelial cells with foci of mild to moderate dysplastic change. In 2 cases, the adjacent pancreas showed foci of intraductal papillary-mucinous adenoma. Histochemically, the tumors largely showed neutral mucin with a lesser amount of acidic mucin made up mainly of sialomucin. Endocrine cells were found in five tumors. Immunohistochemically, all tumors were labeled with M-GGMC-1 and MUC6, whereas MUC1 and MUC2 stains were negative. Pepsinogen II was positive in 5 tumors; thus, the results displayed a pattern of differentiation similar to those of ordinary gastric pyloric or metaplastic pyloric glands. DPC4 expression was maintained in all tumors and p53-positive nuclei were hardly encountered. All patients are alive with no evidence of disease 3 to 10.5 years after surgical resection. PMID- 15832085 TI - Expression of CD163 (hemoglobin scavenger receptor) in normal tissues, lymphomas, carcinomas, and sarcomas is largely restricted to the monocyte/macrophage lineage. AB - CD163, a hemoglobin scavenger receptor, is expressed in monocytes and macrophages. We tested the expression of the CD163 protein in 1,105 human malignancies and normal tissues using tissue microarrays and conventional paraffin-embedded tissue sections. Besides staining nonneoplastic monocytes and histiocytes (tissue macrophages), membranous/cytoplasmic staining for CD163 was primarily limited to neoplasms with monocytic/histiocytic differentiation. CD163 reactivity was not observed in normal tissues, lymphomas, carcinomas, and in a majority of mesenchymal neoplasms, including follicular dendritic cell tumors (0 of 4), although it stained admixed histiocytes. Staining for CD163 was seen in Rosai-Dorfman disease (5 of 6), histiocytic sarcoma (3 of 4), littoral cell angioma (6 of 6), and Langerhans cell histiocytosis (3 of 5). A subset of atypical fibrous histiocytomas (9 of 16), benign fibrous histiocytomas (6 of 9), and atypical fibroxanthomas (1 of 3) also showed CD163 staining. Our studies also confirm earlier work showing that CD163 is expressed in acute myeloid leukemia with monocytic differentiation (AML, FAB subtype M5) (2 of 6), as well as a majority of giant cell tenosynovial tumors (7 of 8). Its limited range of expression and tissue specificity indicate that CD163 may have significant diagnostic utility in separating specific tumors with monocytic and histiocytic derivation from other entities in their differential diagnosis. PMID- 15832086 TI - Cytokeratins in papillary lesions of the breast: is there a role in distinguishing intraductal papilloma from papillary ductal carcinoma in situ? AB - We studied 50 papillary lesions (25 papillomas and 25 papillary ductal carcinomas in situ, DCIS) diagnosed at Singapore General Hospital, for immunohistochemical expression of cytokeratin (CK) 5/6, CK14, and 34betaE12. The immunoscore (proportion of stained cells multiplied by staining intensity) was compared between the two groups. Cytokeratin expression was corroborated by confocal microscopy. Results were applied to a separate series of 43 papillary tumors from Hong Kong (HK). CK5/CK6, CK14, and 34betaE12 showed higher immunoscores in papillomas (mean values, 107.6, 186.6, and 113.1, respectively) than papillary DCIS (mean values, 12, 29.6, and 34.5, respectively; P<0.0001, P<0.001, and P<0.02, respectively). A cutoff immunoscore threshold of 50 appeared discriminatory between papilloma and papillary DCIS, and this value was applied to the HK cases, with CK5/CK6, CK14, and 34betaE12 correctly predicting 25 (89.3%), 26 (92.9%), and 27 (96.4%), respectively, of 28 HK lesions labeled as papillomas; while they corroborated 13 (86.7%), 13 (86.7%), and 5 (33.3%), respectively, of 15 HK cases diagnosed as papillary DCIS. Review of discordant cases showed that lesions were small, derived from core biopsies, or disclosed accompanying invasive carcinoma. When both SGH and HK cases were combined as a group, the sensitivity of an immunoscore of 50 or less in the diagnosis of papillary DCIS was 95%, 85%, and 62.5% for CK5/CK6, CK14, and 34betaE12, respectively, while the specificity was 86.8%, 94.3%, and 86.8%, respectively. CK immunohistochemistry can aid in evaluating papillary breast lesions. 34betaE12 does not appear as useful in identifying papillary DCIS. PMID- 15832087 TI - EGFR mutation is specific for terminal respiratory unit type adenocarcinoma. AB - We have previously reported that terminal-respiratory-unit (TRU) type adenocarcinoma is a distinct subset of lung adenocarcinoma in terms of molecular pathway for carcinogenesis and phenotypic profiles. This type of cancer shows TRU features, characterized by distinct cellular morphology and the expression of TTF 1 and surfactant proteins. Recently, two groups published novel mutations of the epidermal growth factor receptor (EGFR) that are closely associated with clinical response to gefitinib. The clinicopathologic features of gefitinib responders overlap with those of TRU-type adenocarcinoma, and the characteristics of TRU are likely to correspond to the bronchioloalveolar features reported as a predictor of gefitinib response. We therefore examined the characteristics of EGFR-mutated pulmonary adenocarcinomas with special reference to TRU-type adenocarcinoma. EGFR mutation was detected in 97 of 195 adenocarcinomas, 91 of 149 TRU-type adenocarcinomas and 6 of 46 tumors of other types. Conversely, 91 of 97 EGFR mutated adenocarcinomas were categorized as TRU-type adenocarcinomas. This type specific involvement was confirmed by logistic regression model. In addition, EGFR mutation was detected in some cases of atypical adenomatous hyperplasia, a preinvasive lesion of TRU-type adenocarcinoma. These findings further confirm that TRU-type-adenocarcinoma is a distinct adenocarcinoma subset in which a particular molecular pathway is involved. PMID- 15832088 TI - KIT and RCC are useful in distinguishing chromophobe renal cell carcinoma from the granular variant of clear cell renal cell carcinoma. AB - The distinction between chromophobe renal cell carcinoma, the granular cell variant of clear cell renal cell carcinoma, and renal oncocytoma is a common diagnostic dilemma. The usefulness of KIT, CD10, RCC, and RON in the differential diagnosis of these renal epithelial tumors was investigated. KIT was 100% positive in chromophobe renal cell carcinoma (11 of 11) and renal oncocytoma (12 of 12). The KIT staining pattern was identical in both tumor types, with cytoplasmic membrane attenuation, and fine granular cytoplasmic staining. In contrast, KIT was absent in all granular cell variants of clear cell renal cell carcinoma (0 of 6). RCC was observed in more than 80% of the granular cell variant of clear cell renal cell carcinoma (5 of 6) but was negative in all chromophobe renal cell carcinomas (0 of 11) and renal oncocytomas (0 of 12). CD10 was expressed in 100% of the granular cell variant of clear cell renal cell carcinoma (6 of 6), 72% of chromophobe renal cell carcinomas (8 of 11), and 58% of renal oncocytomas (7 of 12). RON was 100% positive in the chromophobe renal cell carcinomas (11 of 11) and renal oncocytomas (12 of 12) but only 50% positive in the granular cell variant of clear cell renal cell carcinoma (3 of 6). Colloidal iron was diffusely and strongly positive in more than 80% of the chromophobe renal cell carcinomas (9 of 11), focally and weakly positive in 41% of the renal oncocytomas (5 of 12) but negative in all granular cell variant of clear cell renal cell carcinoma (0 of 6). The above results demonstrate that: 1) KIT is a very sensitive marker for both chromophobe renal cell carcinoma and renal oncocytoma; 2) immunohistochemistry using antibodies to KIT combined with RCC was sufficient to discriminate between chromophobe renal cell carcinoma and the granular cell variant of clear cell renal cell carcinoma; and 3) neither RON, nor KIT, nor a combination of this panel can be used to distinguish chromophobe renal cell carcinoma from renal oncocytoma. Colloidal iron staining aided in this distinction for the majority of the chromophobe renal cell carcinomas (more than 80% positive) and renal oncocytomas (close to 60% negative). PMID- 15832089 TI - HHV-8-associated T-cell lymphoma in a lymph node with concurrent peritoneal effusion in an HIV-positive man. AB - Primary effusion lymphoma (PEL) is an uncommon large cell lymphoma, usually seen in human immunodeficiency virus (HIV)-infected patients. PEL is characterized by various clinical, histomorphologic, and immunophenotypical features, and is associated with the human herpes virus 8 (HHV-8). PEL may present as either a body cavity-based lymphomatous effusion or a solid tumor mass. Most so-called "solid PEL" usually have an extranodal location; exceptionally rarely, they occur in lymph nodes. The majority of PEL consist of malignant cells of B-cell genotype; seldom they are of T-cell origin. We report a rare case of HHV-8 associated "solid PEL" of T-cell type in a 41-year-old HIV-seropositive man with a concomitant peritoneal effusion. The T-cell lymphoma was diagnosed on the basis of morphologic, immunophenotypic, and molecular findings of a lymph node biopsy. The tumor cells strongly expressed CD45R0, CD7, CD43, MUM1/IRF4, CD30, HHV-8, and EBER, and demonstrated a clonal rearrangement of T-cell receptor-gamma chain gene. The following case provides another example of a lymph node-based "solid" PEL, demonstrating the variety within the spectrum of HHV-8-associated lymphoma. PMID- 15832090 TI - Nuclear beta-catenin expression distinguishes deep fibromatosis from other benign and malignant fibroblastic and myofibroblastic lesions. AB - Deep fibromatoses (desmoid tumors) are clonal myofibroblastic proliferations that are prone to aggressive local recurrences but that do not metastasize. They must be distinguished from a host of fibroblastic and myofibroblastic lesions as well as from smooth muscle neoplasms. Virtually all deep fibromatoses have somatic beta-catenin or adenomatous polyposis coli (APC) gene mutations leading to intranuclear accumulation of beta-catenin. Since low-grade sarcomas in general lack beta-catenin and since reactive proliferations would not be expected to have it, we predicted that nuclear beta-catenin expression would be detected in deep fibromatoses but absent in other entities in the differential diagnosis. We evaluated the role of beta-catenin to help differentiate distinguish deep fibromatoses from congeners. Formalin-fixed, paraffin-embedded sections from 21 lesions from 20 patients with deep fibromatoses were stained with monoclonal beta catenin antibody (Transduction Laboratories) and compared with low-grade fibromyxoid sarcoma (n=12), leiomyosarcoma (n=10), various other fibrosarcoma variants (n=13, including 3 myofibrosarcomas, 3 sclerosing epithelioid fibrosarcomas, 5 low-grade fibrosarcomas, 1 classic fibrosarcoma arising in dermatofibrosarcoma protuberans, 1 inflammatory myxohyaline tumor/myxoinflammatory fibroblastic sarcoma), myofibroma/myofibromatosis (n=12), nodular fasciitis (n=11), and scars (n=9). Nuclear and cytoplasmic staining was assessed. All 21 examples of deep fibromatosis displayed nuclear beta-catenin (focal nuclear staining in one case to 90% staining). All other lesions tested (n=67) lacked nuclear labeling for beta-catenin, showing only cytoplasmic accumulation. beta-Catenin immunohistochemistry separates deep fibromatosis from entities in the differential diagnosis, a finding that can be exploited for diagnosis. Most fibromatoses have diffuse nuclear staining although occasional examples only focally label. PMID- 15832091 TI - Pulmonary adenocarcinomas with enteric differentiation: histologic and immunohistochemical characteristics compared with metastatic colorectal cancers and usual pulmonary adenocarcinomas. AB - Primary pulmonary adenocarcinomas with enteric differentiation (PAED) are mainly composed of tall-columnar cells that show similarity to intestinal epithelia and colorectal carcinomas. In this study, we analyzed the immunostaining profiles of 7 PAEDs in comparison with 14 metastatic colorectal carcinomas (MCRs) and 30 usual pulmonary adenocarcinomas (PACs), using antibodies against CDX-2, cytokeratin 7 (CK7), cytokeratin 20 (CK20), TTF-1, surfactant apoprotein-A (SP A), Napsin A, and MUC2. The positive rates for CDX-2, CK7, CK20, TTF-1, SP-A, Napsin A, and MUC2 were 71%, 100%, 43%, 43%, 14%, 0%, and 43%, respectively, in the PAEDs; 100%, 0%, 86%, 0%, 0%, 0%, and 57% in the MCRs; 3%, 100%, 0%, 93%, 73%, 90%, and 0% in PACs. As expected, immunoreactivity of CDX-2, CK20, and MUC2 was detected in PAEDs. The observed decrease or loss of immunoreactivity for TTF 1, SP-A, and Napsin A indicates that these lesions demonstrate a shift away from their pulmonary phenotype, although CK7 expression was retained. The results indicate that CK7 and CK20 may be useful markers for distinction of PAEDs from MCRs. PMID- 15832092 TI - Genetic aberrations in primary cutaneous large B-cell lymphoma: a fluorescence in situ hybridization study of 25 cases. AB - In contrast to nodal large B-cell lymphomas, recurrent chromosomal aberrations have been studied only in a small number of cases of primary cutaneous diffuse large B-cell lymphoma (PCDLBCL). We investigated 25 PCDLBCLs (classified according to the WHO-EORTC classification into PCDLBCL, leg-type, 8; and PCDLBCL, other, 17), using an interphase fluorescence in situ hybridization technique. All cases were analyzed for chromosomal aberrations commonly observed in nodal large B-cell lymphomas, including structural aberrations of the genes BCL2, BCL6, and c MYC, and numerical aberrations of the chromosomes/genes 3, 7, 8, 11, 12, 13, 17, 18q, RB1, and p53. We observed genetic aberrations in 19 (76%) of 25 patients. The most frequent numerical aberrations were gains of chromosome 12 (7 of 25, 28%), 7 (5 of 25, 20%), 3 (5 of 25, 20%), 18q (3 of 25, 12%), 11 (3 of 25, 12%), X (3 of 25, 12%), and losses of chromosome/gene 17/p53 (3 of 25, 12%). BCL2, c MYC, and BCL6 were rearranged with the IGH gene in 4 (16%), 1 (4%), and none (0%) of 25 cases, respectively. Most aberrations were homogeneously distributed among cases of PCDLBCL, leg-type and of PCDLBCL, other, cases located on the leg or at other body sites, cases with round and cleaved cell morphology, and Bcl-2+ and Bcl-2- cases. These results suggest that PCDLBCLs show similar chromosomal aberrations irrespective of classification, anatomic site, cell morphology, and Bcl-2 expression, and that many similarities between primary cutaneous and nodal diffuse large B-cell lymphomas can be observed. PMID- 15832093 TI - p16INK4A immunoexpression and HPV in situ hybridization signal patterns: potential markers of high-grade cervical intraepithelial neoplasia. AB - Integration of human papillomavirus (HPV) into the cell genome is considered to be an important event in the progression of cervical neoplasia. p16, also a useful biomarker of cervical intraepithelial neoplasia (CIN), shows increased immunoexpression with worsening grades of CIN. This study examines the correlation between p16 immunoexpression, grade of CIN, HPV type, and HPV in situ hybridization diffuse and punctate signal patterns (linked to episomal and integrated viral particles, respectively) in 44 cervical biopsies/LEEP excisions classified as CIN 1 and CIN 2/3. In 22 of 25 (88%) CIN 1 lesions, p16 immunoexpression was confined to the lower half of the epithelium, with sporadic to focal staining in 11 of 25 cases (44%). In CIN 2/3 lesions, 15 of 17 (88.2%) showed diffuse, two-thirds to full-thickness staining of the epithelium. High risk HPV types were found in 20 (80%) CIN 1 lesions and 17 (100%) CIN 2/3 lesions. Punctate signals were detected in only 3 (13.6%) of high-risk HPV positive CIN 1 lesions and in 17 of 17 (100%) CIN 2/3 lesions (P<0.001). p16 immunoexpression and the presence of punctate signal on HPV in situ hybridization correlated with the degree of cervical neoplasia (P<0.001). However, 3 cases of CIN 1 demonstrating punctate signals did not demonstrate a comparable CIN 2/3 p16 staining pattern. Similarly, two CIN 1 lesions with comparable CIN 2/3 p16 staining showed no evidence of viral integration. Both increased p16 immunoexpression and punctate signal correlate with CIN 2/3 grade, supporting the use of either, or both, tests to confirm CIN 2/3. Strong p16 immunostaining in CIN 1 appears independent of HPV punctate signal type. PMID- 15832094 TI - Management of superficial Barrett's epithelium-related neoplasms by endoscopic mucosal resection: clinicopathologic analysis of 27 cases. AB - Endoscopic mucosal resection (EMR), a relatively new endoluminal therapeutic technique with low morbidity and no mortality reported to date, is advocated for the treatment of Barrett's esophagus (BE)-related superficial neoplasms. However, recent studies revise its success downward, particularly regarding the ability to achieve complete excision. To evaluate what remains an evolving technique, we analyzed our experience with a series of 27 esophageal EMRs (20 lesions in 18 patients). Our goal was to evaluate the diagnostic, staging, and therapeutic advantages of EMR separately by correlating the initial biopsies and pre-EMR endoscopic ultrasound (EUS) staging with the final histologic diagnoses and stage. Persistence/recurrence of neoplastic tissue was also correlated with the margin status of the resections. The mean size of the neoplasms, which included low-grade dysplasias (n=2), high-grade dysplasias (n=8), intramucosal carcinomas (n=14), and submucosal invasive carcinomas (n=3), was 11 mm. EUS correctly reported an intramucosal or submucosal lesion in 70% of the cases while it overstaged 18% and understaged 12% of the cases. The biopsy diagnosis corresponded to the EMR diagnosis in 63% of the cases. The biopsy underestimated the grade of the lesion in 21% of the cases. EMR revealed a lower histologic grade compared with the biopsy in 16% of the cases. The resection was microscopically complete in only 4% of the cases. No residual/recurrent disease was observed in 10 lesions (9 patients) at 4 to 63 months (mean, 23 months) post EMR. However, 9 lesions (8 patients) persisted/recurred 28 days to 25 months (mean, 6 months) after treatment; 56% of the cases with positive lateral margin(s) and negative deep margin persisted/recurred. However, 86% of the EMRs with positive deep margin showed residual tumor/recurrence on follow-up biopsies. In conclusion, we observed that EMR offers improved diagnosis and staging as compared with biopsy and EUS. This is a significant advantage since it can modify patients' management. However, frequent incompleteness of resection and high persistence/recurrence are significant pitfalls that dictate continued endoscopic surveillance. PMID- 15832095 TI - B-cell specific activation protein encoded by the PAX-5 gene is commonly expressed in merkel cell carcinoma and small cell carcinomas. AB - PAX-5 is a B cell specific transcription factor crucial for B cell ontogeny and has been detected in most of human B-cell lymphomas. In mouse, PAX-5 is also highly expressed in the central nervous system under tight temporal and spatial controls during embryogenesis. In humans, however, detection of PAX-5 in cells other than B lymphocytes has rarely been reported. We have encountered cases of Merkel cell carcinoma expressing PAX-5 during our routine evaluation of lymphoma. Because Merkel cell carcinoma is a small blue round cell tumor constantly in the differential diagnosis of lymphoma, we expanded our study in an effort to determine if PAX-5 is significantly expressed in neuroendocrine tumors. Based on our immunohistochemistry results using a monoclonal anti-PAX5 antibody with paraffin-embedded tissue sections, we report herein that PAX-5 was detected in 29 of 31 (93.5%) of Merkel cell carcinoma and 22 of 30 (73.3%) of small cell carcinoma, but in none of 17 cases of carcinoid tumor. Furthermore, the staining intensity of PAX-5 in Merkel cell carcinoma was frequently comparable with that in most B-cell lymphomas. We conclude that expression of PAX-5 is not confined to the B cell lineage and is frequently associated with neuroendocrine carcinomas. PMID- 15832096 TI - Incidental and concurrent malignant lymphomas discovered at the time of prostatectomy and prostate biopsy: a study of 29 cases. AB - The incidence and histologic features of malignant lymphomas discovered at the time of prostate biopsy, transurethral resection, and prostatectomy are not well documented. We searched our surgical pathology files for malignant lymphomas identified from prostate surgical specimens from 1989 to 2004. Of 4,831 cases of prostate specimens (3,405 biopsies, 266 transurethral resections, 1,160 prostatectomies) examined at the City of Hope during this period, 29 cases of malignant lymphomas involving the prostate and pelvic lymph nodes were identified (0.6%). These malignant lymphomas can be divided into two groups: 1) 18 incidental cases (0.37%) without prior history of malignant lymphoma; and 2) 11 cases (0.23%) with concurrent known malignant lymphoma. For the first group, the patients with pelvic node involvement ranged in age from 59 to 78 years (mean, 69.2 years; median, 70 years), and the patients with prostate involvement ranged in age from 45 to 78 years (mean, 64.6 years; median, 67.5 years). For the second group, the patients ranged in age from 53 to 80 years (mean, 66.8 years; median, 69 years). Diagnoses of all cases were confirmed by immunohistochemistry or molecular analysis. Ten of 18 cases in the first group involved pelvic nodes only, and the other 8 cases were primary prostatic lymphoma. Of 18 cases in the first group, 13 were small lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL), 3 were marginal zone B-cell lymphoma, and 1 was mantle cell lymphoma. These lymphomas were not readily apparent in most cases by histologic examination, and may be confused with chronic prostatitis when the prostate was involved or with reactive lymphoid hyperplasia when pelvic nodes were involved. Immunohistochemistry and molecular studies may be necessary to confirm the diagnosis. For the second group, prostate and pelvic lymph nodes were involved as part of systemic dissemination of concurrent malignant lymphoma. The diagnosis was usually easily established in these cases. Of 11 cases, 4 were SLL/CLL, 4 were follicular lymphoma, 2 were mantle cell lymphoma, and 1 was diffuse large B cell lymphoma. PMID- 15832097 TI - Primary thyroid teratomas in children: a report of 11 cases with a proposal of criteria for their diagnosis. AB - Cervical teratomas are uncommon neoplasms, although the commonest neck tumors in newborns and infants. Presence of associated thyroid tissue often causes speculation as to the site of origin, ie, arising from within thyroid, adjacent soft tissue with secondary involvement of thyroid, or as innate part of a cervical teratoma. Twenty-eight cases of cervical teratomas were identified over 75 years, including 11 cases containing associated thyroid tissue. Clinical history, treatment, and follow-up were reviewed and the neoplasms analyzed regarding location, size, degree of maturity, and relative arrangement of thyroid and other tissues. All thyroid teratomas were congenital, measured 3.5 to 13.5 cm in diameter (median size, 6.9 cm), and were resected. Follow-up ranged from 1 to 45 years (median, 17 years) without recurrent disease in any patient. Neuroglial tissue predominated in 10 of 11 tumors. Intimate admixture of thyroid and other tissues with or without surrounding fibrous pseudocapsule was present in 8 cases, suggesting thyroid as origin. Histologic immaturity in congenital thyroid teratomas is not the harbinger of adverse behavior as seen in adolescents and adults. Intimate intermingling of thyroid tissue with teratoma and presence of a pseudocapsule seem to be the most significant criteria for establishing thyroid as origin. PMID- 15832098 TI - Factors associated with incidence and persistence of symptoms of disturbed sleep in an elderly cohort: the Cardiovascular Health Study. AB - BACKGROUND: There are limited data pertaining to the factors influencing the incidence and persistence of sleep symptoms in the elderly. The purpose of this study was to determine the incidence and nonremission rates of the following sleep symptoms: trouble falling asleep (TFA), frequent awakenings (FA), and excessive daytime sleepiness (EDS) in the Cardiovascular Health Study (CHS), a prospective multicenter study of cardiovascular disease in a large cohort of elderly adults. Factors influencing these rates were assessed as well. METHODS: 4467 participants in CHS were surveyed for the presence of TFA, FA, and EDS as well as other health problems at their baseline examination and at a follow-up examination 1 to 4 years later. RESULTS: Annualized incidence and nonremission rates were the following: TFA (2.8% and 15.4%), FA (12.3% and 22.7%), and EDS (4.4% and 13.4%). Women were more likely to have incident and persistent TFA. Depression was the primary factor predicting the incidence of all three sleep symptoms. However, other health conditions, including respiratory symptoms and cardiovascular disease, and limitation in activities of daily living were important as well. Depression also was the most important factor associated with persistence of these sleep symptoms. The role of other health conditions in determining nonremission was much more limited. CONCLUSIONS: Incidence of sleep disturbances in the elderly is related to depression, health conditions, and physical functioning. However, persistence of sleep disturbances is best predicted by the presence of depression. PMID- 15832099 TI - High prevalence of C-reactive protein elevation with normal triglycerides (100 149 mg/dL): are triglyceride levels below 100 mg/dL more optimal in coronary heart disease risk assessment? AB - BACKGROUND: The National Cholesterol Education Program defines a fasting triglyceride level below 150 mg/dL as normal. However, observational data suggest that triglyceride levels above 100 mg/dL may predict coronary heart disease (CHD) events. METHODS: To determine the prevalence of systemic inflammation with a normal triglyceride level (100-149 mg/dL), data obtained from the population based cross-sectional study of 4412 men and women in the third National Health and Nutrition Examination Survey were reviewed. Measurements included fasting lipids and lipoproteins and serum C-reactive protein (CRP) level measured using a Behring Nephelometer Analyzer System. RESULTS: High CRP level was equally prevalent whether the designated triglyceride cutpoint was greater than or equal to 100 (unadjusted OR, 2.0; 95% CI, 1.7-2.3) or greater than or equal to 150 (unadjusted OR, 1.9; 95% CI, 1.6-2.2). After adjustment for other covariates, the triglyceride range of 100 to 149 mg/dL remained independently associated with elevated CRP level (OR, 1.3; 95% CI, 1.02-1.67). In addition, an approximately fivefold higher likelihood of elevated CRP level was observed with triglyceride levels between 100 and 149 mg/dL and normal body mass index (BMI; 24-24.9) compared with lower triglyceride level (<65 mg/dL) and BMI (<22) (P < 0.0001). CONCLUSIONS: These data indicate that "normal" triglyceride levels (100-150 mg/dL) are associated with systemic inflammation and that lower fasting triglyceride levels (eg, <100 rather than <150 mg/dL) may be a more optimal cutpoint in CHD risk assessment. PMID- 15832100 TI - Incidence of bacteremia after routine tooth brushing. AB - BACKGROUND: The American Heart Association first developed recommendations on antibiotic prophylaxis against infective endocarditis during dental procedures more than 50 years ago. These recommendations were partly based on the fact that bacteremia occurs with dental procedures. Previous studies in the 1970s and earlier demonstrated that patients become bacteremic after tooth brushing. Improved culture techniques suggest that these rates could be higher now. The objective of this study was to determine the current incidence of bacteremia after routine tooth brushing. METHODS: Thirty military beneficiaries were enrolled in a prospective, institutional review board-approved study after providing informed consent. The incidence of bacteremia after routine tooth brushing for 1 minute using a standardized soft-bristle toothbrush was prospectively measured in 30 healthy adults at three different time points (at baseline and 30 seconds and 20 minutes after brushing). Periodontal Screening and Recording (PSR) scores were recorded for each patient to assess periodontal disease. RESULTS: Three of 180 blood cultures were positive for Propionibacterium acnes (a known contaminant). The remaining blood cultures were all negative. The average PSR score was 9.8 (standard deviation 3.17) for 17 of 30 subjects. CONCLUSIONS: The rate of true bacteremia in this study was zero, which is much lower than previous studies. Bacteremia after tooth brushing in a healthy population is a rare occurrence. Data from previous studies may no longer apply to the current population. Results similar to the ones found in this study during other dental procedures could be an impetus to reevaluate infective endocarditis prophylaxis guidelines. PMID- 15832101 TI - The rationale for pharmacologic therapy in stable chronic obstructive pulmonary disease. AB - The structural changes in airways and alveoli that characterize chronic obstructive pulmonary disease (COPD) result from an abnormal and persistent inflammatory reaction to inhaled noxious particles or gases, notably tobacco smoke. This remodeling of the lung leads to irreversible airflow obstruction. However, COPD should be viewed by clinicians as a treatable condition, since most patients with COPD have an additional reversible component related to increased bronchomotor tone. The use of bronchodilators, especially anticholinergics and beta2-agonists, results in a reduction in airway smooth muscle tone and airflow resistance; this translates into marked improvement in significant clinical outcomes such as dyspnea, quality of life, and exercise capacity. An increasing amount of evidence supports the idea that long-acting agents have more significant impact in these parameters than the short-acting preparations. PMID- 15832102 TI - Ponce de Leon's Fountain: stem cells and the regenerating heart. AB - Despite current pharmacologic and whole organ transplantation strategies, advanced heart failure remains a common and deadly disease. Limited availability of donor organs for use in orthotopic heart transplantation has prompted the examination of alternative therapies, including cell transfer strategies. Stem cell populations have been identified in virtually all postnatal tissues with the exception of the heart, and these stem cells function in the maintenance and regeneration of the respective tissues. Recent studies challenge preexisting notions regarding cardiac repair and suggest that the heart is capable of limited regeneration through the activation of resident cardiac stem cells or the recruitment of stem cell populations from other tissues such as the bone marrow. This review highlights animal models that have the capacity for myocardial regeneration and examines potential sources of stem cell populations that may participate in tissue regeneration. While some authors view these cell-based strategies as a Fountain of Youth for the myopathic heart, future studies will decipher the regulatory mechanisms of stem cell populations and serve as a prelude to stem cell-based strategies. PMID- 15832103 TI - Primary intradural extramedullary hydatid cyst. AB - Spinal hydatid cysts account for 1% of all cases of hydatid disease; primary intradural hydatid cysts are uncommon. We present a case of pathologically confirmed intradural spinal cyst hydatid in an otherwise healthy patient who showed no other evidence of systemic hydatid cyst disease. The patient presented with back pain, paraparesis, and weakness. An intradural extramedullary cystic lesion was identified with magnetic resonance imaging and was shown to be a hydatid cyst by histopathologic examination after surgical removal. To our knowledge, this is the 25th case of hydatid cyst at an intradural extramedullary location reported in the literature. PMID- 15832104 TI - Emphysematous gastritis. AB - Emphysematous gastritis is a rare condition in which gas collects within the stomach wall secondary to infection by gas-producing organisms. The authors report a case of an 82-year-old diabetic man admitted for altered mental status, fever, and leukocytosis. Intramural air within the gastric wall was incidentally found by computed tomography. The case is discussed and differential diagnosis, early diagnosis, and outcome are reviewed. PMID- 15832105 TI - Short stature, type E brachydactyly, exostoses, gynecomastia, and cryptorchidism in a patient with 47,XYY/45,X/46,XY mosaicism. AB - We report a 72-year-old male patient with a 47,XYY/45,X/46,XY mosaicism associated with short stature, exostoses, type E brachydactyly, gynecomastia, cryptorchidism, mild mental retardation, and a paranoid personality and conversion disorder. Since his prevalent cell line was 47,XYY (about 75%), our patient could be karyotypically classified as a case of 47,XYY syndrome. In view of the striking similarity of the clinical features of this case and those of a XYY case previously reported by Ikegawa et al (1992), it seems reasonable to suggest that these patients are representatives of a novel syndrome with a XYY karyotype. PMID- 15832106 TI - Mizoribine-induced rhabdomyolysis in a rheumatoid arthritis patient receiving bezafibrate treatment. AB - Bezafibrate, one of fibric acid derivatives, is widely used to treat hypertriglyceridemia and diabetic dyslipidemia. Fibric acid derivatives are known to induce rhabdomyolysis as a side effect, especially when given to patients with renal dysfunction. Mizoribine, an imidazole nucleoside, is used as an immunosuppressive agent. Here, we present a case of a patient with rheumatoid arthritis who developed rhabdomyolysis while undergoing treatment with mizoribine concomitantly with bezafibrate. Drug-induced rhabdomyolysis was suspected and bezafibrate and mizoribine were discontinued, and the patient was treated with hydration. The patient's symptoms rapidly disappeared and abnormalities of blood and urine test findings also improved to normal levels within 1 week. When prescribing fibrates to patients with high risk of renal damage, caution should be exercised regarding interactions with other drugs and the potential for inducing rhabdomyolysis. PMID- 15832107 TI - Coincidence or causality: celiac and Crohn diseases in a case of Turner syndrome. AB - We describe a 29-year-old woman presenting with chronic diarrhea, growth retardation, and primary amenorrhea who was diagnosed as having celiac disease coexisting with Crohn disease and Turner syndrome. The association of Turner syndrome, inflammatory bowel disease, and celiac disease is reviewed with insights into the genetics of immunologic disorders and possible chromosomal derangements leading to inflammatory bowel disease. To our knowledge, this is the first case of Crohn disease associated with celiac disease and Turner syndrome. PMID- 15832108 TI - From guaiac to immune fecal occult blood tests: the emergence of technology in colorectal cancer screening. AB - Colorectal cancer is the second leading cause of cancer deaths in the United States for both men and women. Colorectal cancer screening is an important means for reducing morbidity and mortality. The American Cancer Society recommends five different screening regimens for adults of average risk, age 50 years and older. The optimal effectiveness of a screening program is dependent on the accuracy of the screening test used. An accurate screening test would have high sensitivity (positive) when an adenomatous polyp or cancer is present and high specificity (negative) in their absence. In April 2002, the American Cancer Society Colorectal Cancer Advisory Group concluded that the immunochemical fecal occult blood test has some advantages that merit revision of their guideline statement for fecal occult blood testing, to include the immunochemical fecal occult blood test. The advantages cited were the possibility of improved sensitivity and specificity and the lack of required dietary restrictions, which make it a more patient-friendly test. Several types of immunochemical fecal occult blood tests are discussed in this article, including their advantages and disadvantages compared with those of the traditional guaiac fecal occult blood testing. PMID- 15832109 TI - Capnography in the gastroenterology lab. AB - This article provides the reader with information and education regarding the use of capnography for both adult and pediatric patients undergoing procedural sedation during endoscopy by: reviewing the basics of capnography, in addition to the physiology of ventilation and oxygenation; illustrating how capnography may provide an earlier warning of hypoxemia than does pulse oximetry; briefly discussing current guidelines for procedural sedation and the potential role of capnography; and reviewing clinical situations causing changes in the capnogram waveforms and the nursing interventions that should be made in response to such changes. PMID- 15832111 TI - An evaluation of bariatric Web sites for patient education and guidance. AB - Bariatric surgery for the treatment of obesity has become a trend, with television and music celebrities touting it as a cure. There has been an extraordinary growth rate in the number of people each year undergoing bariatric surgery, and many of these patients are obtaining their information about the surgery from the Internet. As more and more people turn to the Internet for healthcare information, the need to monitor those Web sites for accuracy and quality expands. The purpose of this study was to assess bariatric Web sites for their quality and accuracy of information. Forty Web sites were evaluated using the Health Information Technology Institute (HITI) criteria and five evaluation criteria based on guidelines from the National Institutes of Health (NIH); the readability of the sites also was determined. Web sites were identified for the purpose of patient education and guidance about this explosive topic. PMID- 15832112 TI - A clarified conceptual meaning of hepatitis C-related depression. AB - A clarified conceptual meaning of depression in individuals with hepatitis C virus infection is proposed based on a critical review of literature. Moving beyond an exclusively biomedical perspective, depression in hepatitis C is explained by a cluster of factors that incorporate physiological, psychological, and social dimensions. Symptom experience, stigma, and uncertainty are factors that span the complex nature of depression in individuals with hepatitis C. This broadened perspective incorporates individual and societal values and beliefs regarding hepatitis C and encompasses the multidimensional complexity of depression in hepatitis C. Hepatitis C-related depression presents an enormous challenge for nurses because of its interference with treatment adherence and significant negative impact on the individual's quality of life. Nursing theorists, researchers, and clinicians may benefit from a clear conceptual understanding of the unique nature of depression in this growing segment of the U.S. population. This clarified conceptual meaning needs to be validated through qualitative, quantitative, and longitudinal studies with this population. It is hoped that future theorists, researchers, and practitioners will contribute to our conceptual understanding, resulting in improved quality of life for this special population. PMID- 15832114 TI - Efficacy and tolerance of sodium phosphates oral solution after diet liberalization. AB - Bowel cleansing regimens commonly require adherence to liquid diets for 24 to 48 hours before examination, which often leads to poor compliance, reduced cleansing, and ultimately inadequate examinations. The authors investigated the efficacy and tolerability of diet liberalization before bowel cleansing with sodium phosphates oral solution. Two hundred patients were randomized into two treatment groups. One group received the standard light breakfast followed by clear liquids the day before colonoscopy; the second had a normal breakfast followed by a low-residue lunch the day before colonoscopy. Both groups had the same bowel preparation with sodium phosphates oral solution (2 x 45-mL, 7 p.m./6 a.m.). There was no difference in clinical efficacy between the two diet regimens (excellent/good in 93% standard, 95% low-residue). Fewer patients receiving the low-residue diet reported hunger, and more patients receiving the low-residue regimen reported energy to perform usual activities. This study supports offering patients a regular breakfast and a low-residue lunch before bowel cleansing with sodium phosphates oral solution. PMID- 15832115 TI - Pictures in the endoscopy suite. PMID- 15832116 TI - Pharmaceutical considerations for patients with dysphagia. PMID- 15832117 TI - Transporting the endoscope. PMID- 15832118 TI - Connectivity. PMID- 15832119 TI - A second chance at life. PMID- 15832120 TI - The need for a national study of colon cleansing procedures. PMID- 15832134 TI - Gait improvement surgery in diplegic children: how long do the improvements last? AB - Gait improvement surgery in ambulatory children with cerebral palsy performed as single-event multilevel surgery is today a well-established modality of treatment, but follow-up studies are lacking. Preoperative and follow-up gait analysis data of 32 diplegic children who underwent single-event multilevel surgery for gait improvement between 1995 and 1998 were evaluated retrospectively. Relevant sagittal plane kinematic parameters of the hip, knee, and ankle joint and time-distance parameters were considered for outcome measures in this study. Postoperative gait analysis was performed three times in all the cases: after discontinuation of the dynamic AFOs (mean 1.0 +/- 0.3 years), after discontinuation of the night splints (mean 2.3 +/- 0.7 years), and at least 1.5 years after discontinuation of physiotherapy and splints (mean 4.4 +/- 1.1 years). The aim of the study was to ascertain whether the improvements in gait function were maintained over these examinations. The authors found that gait function continued to change over 1, 2, and 3 years of follow-up. A general decrease in gait function was measurable in this collective between the first postoperative and the second postoperative evaluations. The results indicate that evaluation of gait improvement surgery in cerebral palsy performed at a minimum of 3 years after surgery would give the most predictive outcome of treatment. PMID- 15832135 TI - Gait in children with cerebral palsy: observer reliability of Physician Rating Scale and Edinburgh Visual Gait Analysis Interval Testing scale. AB - The aim of this study was to test the inter- and intraobserver reliability of the Physician Rating Scale (PRS) and the Edinburgh Visual Gait Analysis Interval Testing (GAIT) scale for use in children with cerebral palsy (CP). Both assessment scales are quantitative observational scales, evaluating gait. The study involved 24 patients ages 3 to 10 years (mean age 6.7 years) with an abnormal gait caused by CP. They were all able to walk independently with or without walking aids. Of the children 15 had spastic diplegia and 9 had spastic hemiplegia. With a minimum time interval of 6 weeks, video recordings of the gait of these 24 patients were scored twice by three independent observers using the PRS and the GAIT scale. The study showed that both the GAIT scale and the PRS had excellent intraobserver reliability but poor interobserver reliability for children with CP. In the total scores of the GAIT scale and the PRS, the three observers showed systematic differences. Consequently, the authors recommend that longitudinal assessments of a patient should be done by one observer only. PMID- 15832136 TI - Measuring physical function in children with spina bifida and dislocated hips: the Spina Bifida Hips Questionnaire. AB - The objective of this study was to develop a questionnaire to evaluate the activities of daily living that are important to children with spina bifida and dislocated hips and their families. Ninety-six items were generated from a review of the literature and interviews with clinicians, parents, and children with spina bifida and dislocated hips. Items were rated independently for "importance" and "severity" by the children and their parents. The 26 most important and severe items were formatted into a self-administered questionnaire, the Spina Bifida Hips Questionnaire (SBHQ). Parents and children completed 2 weeks apart the SBHQ, the Activities Scale for Kids (ASK), and the Pediatric Outcomes Data Collection Questionnaire (PODCQ). The SBHQ showed excellent test-retest reliability (intraclass correlation = 0.97). Construct validity was established by correlations with the ASK (r = 0.79, P < 0.01) and the PODCQ (r = 0.84, P < 0.01). In conclusion, the SBHQ is a valid and reliable questionnaire for the evaluation of treatment outcomes for children with spina bifida and dislocated hips. PMID- 15832137 TI - Postoperative gastric rupture in children with cerebral palsy. AB - Children with cerebral palsy (CP) develop difficulties with swallowing and bowel motility, although the underlying etiology of these disorders is unclear. The authors identified three children treated at their institution in the past 6 years who developed severe gastrointestinal dysfunction leading to gastric rupture after orthopaedic surgery; all three had a history that included CP (subtype spastic quadriparesis), low cognitive function, and gastrointestinal motility disorders. All three cases were fatal; in two patients the diagnosis was made at autopsy. No case of postoperative gastric rupture has been identified in children undergoing orthopaedic surgery without concomitant CP at the authors' institution. In this case series, they describe the preoperative and postoperative course of these three children. Since the only cure for gastric rupture is prompt surgical attention, orthopaedists should consider gastric rupture in the differential diagnosis of a postsurgical CP patient who becomes acutely unstable. PMID- 15832138 TI - Is operative blood loss associated with valproic acid? Analysis of bilateral femoral osteotomy in children with total involvement cerebral palsy. AB - Valproic acid (VPA) may increase surgical bleeding. The purpose of this study was to review the effect of VPA on blood loss during bilateral femoral osteotomy in children with cerebral palsy. The medical records of 29 children were retrospectively reviewed. Blood volume, total blood loss, and postoperative blood loss were calculated. Fourteen children were taking no anticonvulsant medications, nine children were taking anticonvulsant medications other than VPA, and six children were taking anticonvulsant medications including VPA. The group taking anticonvulsant medications including VPA had significantly lower preoperative platelet counts (204,000) and a significantly greater incidence of transfusion (50%). Compared with the group taking no anticonvulsant medications, the group taking anticonvulsant medications including VPA had a significantly greater change in hematocrit and greater blood loss. Blood loss and the need for transfusion should be anticipated for in children with cerebral palsy taking VPA who undergo femoral osteotomy. PMID- 15832139 TI - Accuracy of intramuscular injection of botulinum toxin A in juvenile cerebral palsy: a comparison between manual needle placement and placement guided by electrical stimulation. AB - Most clinicians who perform botulinum toxin A injections for children with cerebral palsy do so using the "free-hand" or manual technique without using radiologic or electrophysiologic guidance to aid needle placement. The objective of this study was to investigate the accuracy of manual needle placement compared with needle placement guided by electrical stimulation. A total of 1,372 separate injections for upper and lower limb spasticity were evaluated in 226 children with cerebral palsy. The accuracy of manual needle placement compared with electrical stimulation was acceptable only for gastroc-soleus (>75%); it was unacceptable for the hip adductors (67%), medial hamstrings (46%), tibialis posterior (11%), biceps brachii (62%), and forearm and hand muscles (13% to 35%). The authors recommend using electrical stimulation or other guidance techniques to aid accurate needle placement in all muscles except the gastroc-soleus. Further study is needed to determine whether more accurate injecting will lead to better functional outcomes and more efficient use of botulinum toxin A. PMID- 15832140 TI - The diagnosis and management of neonatal hip instability: results of a clinical and targeted ultrasound screening program. AB - This article reports the results of a neonatal hip screening program comprising clinical screening and targeted ultrasound performed by orthopaedic surgeons. Over 7 years, from 1995 to 2001, there were 15,397 live births in the authors' region. Seven hundred thirty-three babies (4.8% of births) were referred for hip ultrasound: 80% for risk factors and 20% for instability. Eighty-three babies (5.4/1,000) were splinted in a Pavlik harness. Three of these subsequently required surgery (1.9/1,000). Ten patients (0.65/1000) presented with hip dislocation after 12 weeks of age, nine of whom required open or closed reduction (0.56/1,000). From 1978 to 1985, when neonatal pediatricians clinically screened all babies, 18 babies presented late from 13,707 births (1.3/1000). Since the introduction of orthopaedic screening and targeted ultrasound, there has been a significant reduction in late diagnosis in the authors' institution. PMID- 15832141 TI - Residual shortening after Legg-Calve-Perthes disease, focusing on the response of the ipsilateral tibia. AB - Residual shortening of the affected limb was measured at skeletal maturity by teleoroentgenograms in 68 patients with Legg-Calve-Perthes disease (LCPD); special attention was paid to the length of the ipsilateral tibia. Of these 68 patients, 38 were treated by abduction orthosis (AO) and 30 by femoral varus osteotomy (FVO). Residual shortening in AO group was significantly greater than that in FVO group. The femoral lengths in both of these groups were similar (12.5 mm in the AO group and 10.1 mm in the FVO group), but the tibial lengths were significantly different (2.5 mm shortening in the AO group and 0.9 mm lengthening in the FVO group). Residual shortening in the patients treated by FVO was less than that in patients treated by AO. The difference is speculated to be caused by the overgrowth of the ipsilateral tibia. PMID- 15832142 TI - Developmental pattern of femoral shortening following devascularization of the capital femoral epiphysis in piglets. AB - To investigate the developmental pattern of femoral shortening in Legg-Calve Perthes disease, the authors made an experimental model of the disease in piglets by devascularizing the capital femoral epiphysis and following the piglets to skeletal maturity. The discrepancy first increased in the postoperative 0 to 8 weeks (1.2-1.9 mm of shortening per week), then decelerated for a considerable period during the postoperative 8 to 16 weeks (0.2-0.6 mm per week), and then increased again toward the end of the growth period of the postoperative 20 weeks (1.2 mm per week). The developmental pattern of femoral shortening showed an upward slope/plateau/upward slope pattern, as in type IV of the Shapiro classification. As the mechanism of the observed pattern, the authors presumed reduced epiphyseal height and growth retardation in the physis during the initial upward slope, a repair process at the plateau phase, and premature physeal closure during the second upward slope. PMID- 15832143 TI - Anatomic study of femoral patellar groove in fetus. AB - The authors performed a biometric analysis of the femoral patellar groove in fetus and compared their findings with those observed in adults. Forty-four formalin-preserved fetuses were studied (13-38 weeks). Digitalized images were used to obtain measurements (alpha angle of the groove, trochlear slopes thetaL and thetaM). A comparison of means of independent samples between our series and adults was performed. For each angle of the distal epiphysis (alpha, thetaL, thetaM) there was no significant difference between this fetal series and adults. This is the first biometry of the fetal patellar groove. The morphology of the lower femur appears to be the same in fetus and adults. The results of this study suggest that the anatomic characteristics of the patellar groove could have been integrated into the genome during the course of evolution. This would be in favor of a genetic origin of patellar groove dysplasia. PMID- 15832144 TI - Treatment of pediatric both-bone forearm fractures: a comparison of operative techniques. AB - Both-bone forearm fractures of the radius and ulna are a common injury in children. Closed reduction and casting has historically been the primary means of treatment in over 90% of these fractures. Unstable and irreducible fractures, however, often pose a therapeutic challenge, with little data available to compare outcomes. The authors performed a retrospective review of 50 children with both-bones fractures treated with closed reduction and cast immobilization, open reduction and internal fixation (ORIF), or intramedullary (IM) nailing. Complications were tabulated and separated by treatment modality and subdivided into minor/major complications. Statistical regression was performed. There were 54 operations in 50 patients with both-bones fractures. All fractures healed within 8 to 10 weeks, except for two delayed unions and one nonunion. The complication rate was 5% for closed treatment, 33% for ORIF, and 42% for IM nailing. Complication rates were significantly different between the closed and operative groups. When comparing treatments in pediatric both-bones fractures, there are significantly more complications with operative techniques. Patients with ORIF had more major complications, often requiring a return to the operating room. IM nailing, when done correctly, is as acceptable and safe a form of treatment. PMID- 15832145 TI - The effectiveness of physiotherapy after operative treatment of supracondylar humeral fractures in children. AB - The indications for physiotherapy after supracondylar humeral fractures in children are not clear in the literature, even in the presence of an active or passive limitation of elbow joint motion. The authors therefore performed a prospective randomized study to assess the effectiveness of physiotherapy in improving the elbow range of motion after such fractures. The authors studied two groups of 21 and 22 children with supracondylar humeral fractures Felsenreich types II and III, all without associated neurovascular deficits. All children were treated by open reduction and internal fixation with Kirschner wires inserted from the radial side of the humerus. Postoperative follow-up at 12 and 18 weeks showed a significantly better elbow range of motion in the group with weekly physiotherapy, but there was no difference in elbow motion after 1 year. In each group, one child had an extension deficit of 15 or 20 degrees. The authors conclude that postoperative physiotherapy is unnecessary in children with supracondylar humeral fractures without associated neurovascular injuries. PMID- 15832146 TI - Posterior divergent dislocation of the elbow in children and adolescents: a report of three cases and review of the literature. AB - Divergent dislocation of the elbow is defined as a specific elbow dislocation in which the distal humerus is forced between the proximal radius and ulna, resulting in separation or divergence of the proximal forearm bones. This is an uncommon injury in children, with 14 cases reported in the last 23 years. The authors report three additional cases and the findings from cadaver elbow dissections in which the injury was reproduced. The authors propose that there is only one type of divergent dislocation, resulting from indirect forces transmitted to the elbow from a fall on the outstretched hand. The injury predominantly occurs in younger children because joint laxity is a predisposing cause. Early recognition, followed by a closed reduction and a 3- to 4-week period of cast immobilization, should result in a full recovery for most children. PMID- 15832147 TI - Buckle fractures of the distal radius are safely treated in a soft bandage: a randomized prospective trial of bandage versus plaster cast. AB - To show that the treatment of buckle fractures of the distal radius in children in a soft bandage is an effective and safe method of treatment, a randomized prospective trial was undertaken. Patients entered the trial after diagnosis in the accident and emergency department. Allocation to either plaster cast or bandage was random. Bandage patients were seen each week. Plaster cast patients were seen at 4 weeks. Measurements were taken at all visits. Patient questionnaires were completed at the end of treatment. Thirty-nine patients completed the study. Eighteen were allocated to bandage, 21 to plaster cast. Those in bandage showed an excellent range of movement by the first week. Results were highly positive for treatment in bandage, with no reported adverse effects and a highly desirable result for the patient. The authors would suggest a change in treatment policy for such fractures. PMID- 15832148 TI - Oral bisphosphonates to treat disuse osteopenia in children with disabilities: a case series. AB - Children with congenital conditions who are nonambulatory have been observed to have pathologic fractures due to disuse osteopenia. Data support the use of intravenous bisphosphonates to treat this in children with disabilities, but there are no data to guide the use of oral bisphosphonate medication. Ten nonambulatory children with disuse osteopenia secondary to either static brain injury or spina bifida were started on alendronate. Investigators completed a chart review of each subject. There were 17 fractures before starting alendronate and 1 fracture in the follow-up period. All children with nociceptive behavior or pain complaints (5/10) had their symptoms resolve after treatment. Despite the presence of gastrointestinal comorbidities (7/10 with a history of gastroesophageal reflux disease, 5/10 taking reflux medication, and 4/10 with PEG tubes), only 1 of the 10 children discontinued the medication secondary to gastrointestinal complaints. This case series offers evidence that disabled nonambulatory children tolerate alendronate, and it may decrease fractures in those at risk from severe disuse osteopenia. PMID- 15832149 TI - Bone healing in children with osteogenesis imperfecta treated with bisphosphonates. AB - The long-term effects of bisphosphonate treatment in children with osteogenesis imperfecta (OI) are unknown. The aim of this study was to evaluate whether treatment with bisphosphonates interferes with the healing of fractures in a group of children with OI. Seven subjects (6 boys), aged 11.4 +/- 5.95 years, were followed for 2.5 +/- 0.84 years after the start of treatment with intravenous pamidronate (9 mg/kg/y) and/or oral alendronate (5 or 10 mg/d). Orthopaedic surgery of 24 bones was performed after 2.33 +/- 4.14 months of treatment, with 1.6 +/- 0.84 osteotomies per bone. Ambulation was started after 26.1 +/- 32.28 days. Reoperation was required in 8% of the bones due to fracture below primary fixation. Pseudoarthrosis was seen in one fracture, an osteotomy of the proximal femur (14% of the patients, as expected in an OI population). These results suggest that treatment with bisphosphonates at the administered doses does not interfere with fracture healing. Larger and longer studies are warranted. PMID- 15832150 TI - The effect of temporary hemiepiphyseal stapling on the growth plate: a radiologic and immunohistochemical study in rabbits. AB - The purpose of this study was to investigate the effect of temporary hemiepiphyseal stapling on the bone geometry and proliferative activity of the physis in immature rabbits. Proximal medial epiphyseal stapling of the right tibia was performed in 46 6-week-old New Zealand white rabbits. The rabbits were assigned randomly into two groups. In group 1, the staples were inserted extraperiosteally and the rabbits were killed at the end of 3 weeks. In group 2, the staples were fixed subperiosteally (group IIA) or extraperiosteally (group IIB), the staples were removed at the end of 3 weeks, and the rabbits were killed at the end of 6 weeks. The articular line-diaphysis angle (ALDA) was significantly increased with 3 weeks of stapling. After the removal of staples, while ALDA continued to worsen in group IIA, it improved in group IIB. Bone was observed to bridge the physis in group IIA. However, the proliferative activity of the physis continued. Temporary hemiepiphyseal stapling is a safe and effective method for control of physeal growth of long bones before skeletal maturity. However, it is of paramount importance not to disturb the periosteum during stapling. PMID- 15832151 TI - Transphyseal bioabsorbable screws cause temporary growth retardation in rabbit femur. AB - A self-reinforced bioabsorbable poly-L-lactide/polyglycolide (SR-PLGA) 80/20 screw 2.0 mm in diameter was implanted transphyseally across the distal growth plate of the right femur in 24 immature rabbits. Radiologic evaluation revealed a mean shortening of 3.1 mm at 3 weeks, 11.1 mm at 6 weeks, 9.3 mm at 24 weeks, 9.0 mm at 48 weeks, and 12.6 mm at 72 weeks compared with the intact contralateral femur. In 13 control rabbits, drilling without screw placement did not cause any statistically significant femoral shortening. Therefore, the transphyseal SR-PLGA 80/20 screw caused growth retardation for 6 weeks postoperatively, after which the normal growth tendency was recovered until the growth plate was closed. The duration of temporary growth retardation correlated with that of strength retention of the SR-PLGA 80/20 copolymer. These findings suggest that SR-PLGA 80/20 screws can be applied in transphyseal bone fixation. The use of bioabsorbable screws for temporary epiphysiodesis seems attractive but requires further study. PMID- 15832152 TI - Disseminated thrombosis-induced growth plate necrosis in rat: a unique model for growth plate arrest. AB - Exposure of rats to 2-butoxyethanol (BE) produces early hemolytic anemia and disseminated thrombosis. This leads to infarctions in multiple organs, including bones and cartilage. BE, administered for different durations of exposure in two separate experiments, produced metaphyseal vascular thrombosis, growth plate infarction, and partial or complete physeal growth arrest. This reproducible model may serve as a useful tool in the study of some conditions that manifest growth plate damage. The suitability of this model for investigating the pathogenesis of growth plate necrosis and as a model for potential therapy for various human growth plate disorders are discussed. PMID- 15832153 TI - Demographics of idiopathic clubfoot: is there a seasonal variation? AB - A seasonal variation in the incidence of idiopathic clubfoot has been reported. The purpose of this study was to determine whether a seasonal variation existed in the authors' population. The medical records of 245 children who presented with idiopathic clubfoot were reviewed. Expected month of birth, month of conception, and season of birth were determined. National Vital Statistics Reports for Births: Final Data for 2001 provided comparison data. The authors' group differed from the national data, with a male predominance and a greater percentage of mothers less than 25 years old at delivery, but was similar in regard to infants born preterm, cesarean delivery, and low birthweight. Analysis failed to identify a monthly or seasonal variation in the authors' population. PMID- 15832154 TI - Use of a soft tissue expander before surgical treatment of clubfoot in children and adolescents. AB - Primary closure of the skin after surgical correction of severe clubfoot is difficult, sometimes impossible. The authors describe a surgical technique with the use of a soft tissue expander prior to surgical treatment on clubfoot in children and adolescents. They operated on 13 feet (10 patients) with rigid clubfoot, with ages ranging from 3 to 16 years (average 10 years). Satisfactory skin healing with primary closure of the skin was obtained in 10 feet; the remaining 3 had complications such as infection, skin necrosis, and premature exposure of the expander. The authors conclude that soft tissue expansion before surgical correction of clubfoot in selected cases is a good alternative for the management of skin closure. Special attention should be given to surgical technique. PMID- 15832155 TI - Flexor digitorum accessorius longus muscle is associated with familial idiopathic clubfoot. AB - The etiology of idiopathic and familial clubfoot remains elusive. The purpose of this study was to document the prevalence of the flexor digitorum accessorius longus muscle in a large series of children treated with an extensive soft tissue release for idiopathic clubfoot deformity and compare those patients with and without this anomalous muscle in terms of various demographic data, including a positive family history of clubfoot. A retrospective review was conducted to identify infants treated with an extensive soft tissue release for idiopathic clubfoot deformity at the authors' institutions between 1980 and 2000. Patient characteristics and family demographic data were analyzed using multiple logistic regression analysis modeling the probability of having the anomalous flexor muscle. A flexor digitorum accessorius longus muscle was identified in 55 (6.6%) of the 835 patients at the time of surgical correction of the clubfoot deformity. It was present in 4.5% of patients without a family history (33/741) and 23.4% of patients with a family history (22/94) (P < 0.0001). Children with first-degree relatives with clubfoot are 6.6 times more likely to have the anomalous flexor muscle than children without first-degree relatives with clubfoot (odds ratio 6.6; 95% confidence interval 3.63-11.84; P < 0.0001). PMID- 15832156 TI - Correction of cavovarus foot deformity in Charcot-Marie-Tooth disease. AB - Operative correction of cavovarus foot deformity in Charcot-Marie-Tooth disease (CMT) is challenging. This progressive peripheral sensory and motor neuropathy commonly involves the forefoot, midfoot, hindfoot, and toes. The authors present a new imaging technique that allows the surgeon to assess the flexibility of the hindfoot in patients with CMT to determine the best operative procedure to correct the deformity. Twenty-five patients (41 feet) with CMT and cavovarus foot deformity were evaluated and a new radiographic technique was studied in some of these patients to determine the usefulness of this imaging modality. The authors believe that this new imaging method will aid in determining the optimal operation for correcting this complex deformity. PMID- 15832157 TI - Buoyancy of synthetic cast and its implications to young patients. AB - One of the authors' young patients suffered near-drowning while being treated with a synthetic below-knee cast, and this intrigued them into examining the buoyancy of waterproof casts and their use in small children. Synthetic and fiberglass casts were fabricated and subjected to immersion testing using lead pellets. Fiberglass casts sank immediately, while synthetic casts stayed afloat. With the addition of weights averaging 221 g, the synthetic casts finally sank. When supine, the casts stayed afloat with more weights. This study shows that synthetic liners are buoyant. Although the weight necessary to sink the casts was small, the weights were lead pellets, which is different from a human limb. Enhanced buoyancy of synthetic casts may pose a potential danger to a small child when not properly supervised in water. The initial buoyant force may be too startling and may place them at a higher risk for near-drowning. PMID- 15832158 TI - Hereditary multiple exostosis and pain. AB - This study was undertaken to characterize pain in individuals with hereditary multiple exostosis (HME). Two hundred ninety-three patients with HME completed a questionnaire designed to assess pain as well as its impact on their life. Eighty four percent of participants reported having pain, indicating that pain is a real problem in HME. Of those with pain, 55.1% had generalized pain. Two factors were found to be associated with pain outcome: HME-related complications and surgery. Individuals who had HME-related complications were five times more likely to have pain, while those who had surgery were 3.8 more likely to have pain. No differences were found between males and females with respect to pain, surgery, or HME-related complications. The results of this study indicate that the number of individuals with HME who have pain has been underestimated and that pain is a problem that must be addressed when caring for individuals with HME. PMID- 15832159 TI - Pre-centralization soft tissue distraction for Bayne type IV congenital radial deficiency in children. AB - There are several surgical options for the treatment of severe congenital radial deficiency. The aim of the current study was to evaluate the results of a staged protocol consisting of gradual soft tissue distraction with an Ilizarov external fixator followed by centralization of the ulna. Three patients (four extremities) with Bayne type IV radial club hand were treated at an average age of 18 months. At a follow-up of 26 months, an average correction of 72 degrees in the hand forearm angle and an improvement of 19 mm in the hand-forearm position were obtained. There were no cases of neurovascular compromise, wound infection, carpal resection, or ulnar shortening. All families were pleased with the results. These findings suggest that a staged surgical approach is a technically feasible alternative in the treatment of severe congenital radial club hand deformity in young children. PMID- 15832160 TI - Two-stage flexor tendon reconstruction in zone 2 of the hand in children. AB - Two-stage flexor tendon reconstruction (Hunter) is indicated in children with extensive adhesions in zone 2 of the hand, with some reservations concerning the patient's age and cooperation. Nine children (mean age 6.9 years) were treated with the modified Paneva-Holevich technique, which has advantages over the classic Hunter reconstructions. It involves an intrasynovial graft (FDS of the injured finger) that is anatomically stable and morphologically more appropriate compared with free grafts. The size of the silicone rod is precisely assessed in the first stage, the proximal tenorrhaphy has healed by the time the second stage is performed, and donor site morbidity is minimized. After a mean follow-up of 40.1 months, the mean total active motion was 196 degrees, and eight patients achieved a good or excellent result according to the Buck-Gramcko and the revised Strickland scale. Staged flexor tendon reconstruction is technically feasible even in very young children. Results in children are comparable to those achieved in adults. PMID- 15832161 TI - Modern surgical treatment of primary aneurysmal bone cyst of the spine in children and adolescents. AB - Aneurysmal bone cyst (ABC) is a benign but locally aggressive tumor-like condition. Treating children with spinal involvement poses special hazards due to the proximity of the lesion to the spinal cord and the need to preserve spinal stability and balance after surgery. Twelve children with ABC treated between 1990 and 2002 at a tertiary pediatric musculoskeletal tumor center using modern surgical techniques and technology were retrospectively reviewed. A surgical technique using a four-step approach of intralesional curettage, high-speed bur, electrocautery, and bone grafting was found to have a significantly reduced rate of recurrence (0/8 cases) compared with traditional intralesional curettage and bone grafting (4/4 cases) (P < 0.002). Short-segment spinal fusion with instrumentation was also done in five cases immediately after excision of the ABC under the same anesthesia and was a nonsignificant marker for reduced rate of recurrence (P < 0.08), likely secondary to wide exposure. Overall, at last follow up, all 12 patients were alive and well with no evidence of disease and no significant spinal deformity. The four-step approach to treatment of ABC of the spine with appropriate spinal instrumentation and fusion is recommended for successful treatment of this aggressive lesion. PMID- 15832162 TI - The contribution of hospital volume, payer status, and other factors on the surgical outcomes of scoliosis patients: a review of 3,606 cases in the State of California. AB - While volume/outcomes relationships have been shown for several areas of orthopaedics, previous studies have not examined this relationship in the area of scoliosis surgery. The Office of Statewide Planning and Development (OSHPD) California inpatient discharge database was used for a retrospective review of all patients 25 years of age or younger with a diagnosis of scoliosis and a spinal fusion procedure from 1995 to 1999 (n = 3,606). Univariate and multivariate analyses were conducted to determine the effect of various factors on in-hospital mortality, surgical complications, reoperations, and length of stay (LOS). Univariate analyses revealed significant effects of age, sex, illness severity, neuromuscular disease, surgical approach, Medicaid status, and annual hospital volume on outcomes (P < 0.05). After controlling for these factors using multivariate regression, patients insured by Medicaid were found to have a significantly greater odds for complications (P = 0.017) and a significantly increased LOS (P < 0.001) compared with patients with all other sources of payment. Additionally, multivariate regression revealed an inverse relationship between annual hospital volume and likelihood of reoperation, as patients treated at hospitals with annual volumes of 5.1 to 25.0, 25.1 to 50.0, and greater than 50.0 spinal fusions all had approximately half the odds of reoperation (P = 0.042, P = 0.004, and P = 0.028 respectively) as patients treated at hospitals with an annual volume of 5.0 or fewer spinal fusions per year. The current data suggest that being insured with Medicaid in the state of California is associated with poorer outcomes after scoliosis surgery. Additionally, this study documents a volume/outcomes relationship in scoliosis surgery. PMID- 15832163 TI - Assessment of children with brachial plexus birth palsy using the Pediatric Outcomes Data Collection Instrument. AB - The purpose of this study was to determine whether the Pediatric Outcomes Data Collection Instrument (PODCI) measures differences in function between children with brachial plexus birth palsy (BPBP) who are candidates for shoulder tendon surgery and age-matched controls. The PODCI was administered prospectively to 23 children with BPBP who were candidates for shoulder tendon surgery. Their results were compared with published PODCI data for control subjects, and factors associated with function within the BPBP cohort were determined. Children in the BPBP cohort had significantly lower PODCI scores in upper extremity function, sports, and global function than control subjects. Limited active shoulder external rotation was significantly associated with lower functional scores. The PODCI measures diminished upper extremity function in children with BPBP who are candidates for shoulder tendon surgery, thereby showing promise as a tool for measuring baseline function and postoperative functional gains for children with BPBP. PMID- 15832164 TI - The Pediatric Outcomes Data Collection Instrument (PODCI) and functional assessment of patients with unilateral upper extremity deficiencies. AB - The Pediatric Outcomes Data Collection Instrument (PODCI) questionnaire was used to quantify functional abilities of a group of unilateral upper extremity deficiency (U-UED) patients and compare them with "normal" control children. Sixty-four consecutive patients with U-UED were assessed. Parents and adolescent (ages 11-21) patients responded. Underlying diagnosis, amputation level, and type of prosthesis were recorded. Scores were compared for congenital versus traumatic etiologies for patients with various amputation levels, and for patients using prostheses versus those not using prostheses. In both parent and patient responses, PODCI scores were significantly lower than "normal" for upper extremity function and sports. Scores were similar for congenital and acquired amputees. Responses from adolescent patients showed progressively decreasing scores for upper extremity, transfers, sports, and global function with progressively proximal amputation levels. Patients using prostheses with different terminal devices did not significantly differ. Parent responses for prosthesis wearers showed lower comfort/pain scores (ie, increased pain) than non prosthesis wearers, but no significant differences in function, including upper extremity function. PMID- 15832165 TI - Is it safe to use chronological age in leg length discrepancy? PMID- 15832166 TI - Limb lengthening for stature: a second opinion. PMID- 15832168 TI - Osteogenesis imperfecta presenting as simultaneous bilateral tibial tubercle avulsion fractures in a child. PMID- 15832169 TI - The Lewis blood group--a new genetic marker of obesity. AB - In a cross-sectional epidemiological study including 3290 men aged 53-75 y, mean=63, we tested the hypothesis that the Lewis phenotype Le(a-b-) is a genetic marker of obesity. All men were Lewis typed and measurements were made of height, weight, neck circumference, and hip fat fold. Obesity was defined as a body mass index >/=30 kg/m(2). Totally 291 men (8.8%) were obese; 9.6% of the men had the Le(a-b-) phenotype. Le(a-b-) men had a higher prevalence of obesity than others, 15.6 vs 8.1%, odds ratio (95% confidence interval): 2.1(1.5-2.9), P<0.001. The etiological fraction, that is, the excess prevalence of obesity in the study population due to the Le(a-b-) phenotype, was approximately 10%. The frequency of the Le(a-b-) phenotype may vary substantially in different populations. Identification of this new genetic marker of obesity may, for example, contribute to the explanation of individual and ethnic differences in the prevalence of obesity. PMID- 15832170 TI - The histone-binding code of nuclear receptor co-repressors matches the substrate specificity of histone deacetylase 3. AB - Ligands for nuclear receptors facilitate the exchange of co-repressors for coactivators, leading to chromatin modifications that favour the activation of gene transcription. Here, we show that the repressed state of an endogenous retinoic acid-regulated gene is quickly re-established after ligand removal. As expected, repression is characterized by recruitment of N-CoR/SMRT-HDAC3 (histone deacetylase 3) co-repressor complexes, leading to local histone hypoacetylation. The achievement of the repressed state involves the ordered deacetylation of lysines in H4 tails. This order is determined by the inherent substrate specificity of HDAC3, and unexpectedly predicts the binding preference of N CoR/SMRT for submaximally acetylated H4 tails. The match between the specificity of acetyl-histone deacetylation by HDAC3 and the histone-binding preference of N CoR/SMRT allows the co-repressor complex to stabilize and propagate repression of nuclear hormone receptor gene targets. PMID- 15832171 TI - Woodchuck post-transcriptional element induces nuclear export of myotonic dystrophy 3' untranslated region transcripts. AB - The woodchuck post-transcriptional regulatory element (WPRE) can naturally accumulate hepatitis transcripts in the cytoplasm, and has been recently exploited as an enhancer of transgene expression. The retention of mutant myotonic dystrophy protein kinase (DMPK) transcripts in the nucleus of myotonic dystrophy (DM) cells has an important pathogenic role in the disease, resulting in pleiotropic effects including delayed myoblast differentiation. In this study, we report the first use of WPRE as a tool to enhance nuclear export of an aberrantly retained messenger RNA. Stable cell lines expressing the normal and mutant DMPK 3' UTR (3' untranslated region) complementary DNA, with or without WPRE, were produced. It is noteworthy that WPRE stimulated extensive transport of mutant transcripts to the cytoplasm. This was associated with repair of the defective cellular MyoD levels and a subsequent increase in myoblast differentiation. These results provide the basis for a cellular model that can be exploited in DM and in the study of RNA transport mechanisms. PMID- 15832172 TI - Oncolytic adenovirus CG7870 in combination with radiation demonstrates synergistic enhancements of antitumor efficacy without loss of specificity. AB - Conditionally replicating adenoviruses that selectively replicate in tumor cells, but not in normal cells, are being explored as virotherapeutic agents for cancer. A prostate-specific oncolytic adenovirus, CG7870 is currently being evaluated in phase 1/2 clinical trials for the treatment of prostate cancer. To decrease the effective dose and further increase the therapeutic efficacy of CG7870, the combination of virotherapy with radiation therapy was explored in this study. CG7870 is an oncolytic adenovirus in which tumor-specific promoters are driving the expression of E1A and E1B proteins. The effects of combined treatment with CG7870 and radiation on cultured cells were determined in cytotoxicity and virus yield assays. The antitumor efficacy of CG7870 (1 x 10(7) particles/mm3 of tumor), 10 Gy of local radiation or both was evaluated in established subcutaneous LNCaP xenografts in nude mice. In vitro, the dual agent treatment resulted in synergistically enhanced potency at suboptimal doses of radiation and virus. Virus yield in irradiated cells increased relative to yield in nonirradiated cells without compromising the specificity of the vector for its target cell types. In vivo, CG7870 treatment alone suppressed tumor growth and extended tumor nonprogression time. The average tumor-volume of the groups treated with CG7870 only and radiation only was 121 and 126% of baseline, respectively, 39 days after treatment. The average tumor-volume of the combination group was 34% of baseline 39 days after a single dose of treatment. No significant body weight loss was observed in any treatment group. There was a significant drop in serum level of prostate-specific antigen (PSA) in the combination group compared to the group treated with either agent alone. In mice treated with CG7870 only or radiation only, serum PSA levels changed to 26 and 383% of baseline, respectively, by study day 46. In contrast, PSA levels in mice treated with CG7870 plus radiation decreased to less than 11% of baseline by study day 46. Histological analysis of tumor sections collected from the combination group revealed enhanced necrosis and more apoptotic cells. Combination of CG7870 with radiotherapy significantly increased antitumor efficacy compared to either agent alone. These results suggest that CG7870 in combination with radiation has improved antitumor efficacy at lower doses and with no additional side effects. PMID- 15832173 TI - Epidermal growth factor receptor-transfected bone marrow stromal cells exhibit enhanced migratory response and therapeutic potential against murine brain tumors. AB - We have created a novel cellular vehicle for gene therapy of malignant gliomas by transfection of murine bone marrow stroma cells (MSCs) with a cDNA encoding epidermal growth factor receptor (EGFR). These cells (EGFR-MSCs) demonstrate enhanced migratory responses toward glioma-conditioned media in comparison to primary MSCs in vitro. Enhanced migration of EGFR-MSC was at least partially dependent on EGF-EGFR, PI3-, MAP kinase kinase, and MAP kinases, protein kinase C, and actin polymerization. Unlike primary MSCs, EGFR-MSCs were resistant to FasL-mediated cytotoxicity and were capable of stimulating allogeneic mixed lymphocyte reaction, suggesting EGFR-MSCs possess suitable characteristics as vehicles for brain tumor immuno-gene therapy. Following injection at various sites, including the contralateral hemisphere in the brain of syngeneic mice, EGFR-MSCs were able to migrate toward GL261 gliomas or B16 melanoma in vivo. Finally, intratumoral injection with EGFR-MSC adenovirally engineered to secrete interferon-alpha to intracranial GL261 resulted in significantly prolonged survival in comparison to controls. These data indicate that EGFR-MSCs may serve as attractive vehicles for infiltrating brain malignancies such as malignant gliomas. PMID- 15832174 TI - Is Apaf-1 expression frequently abrogated in melanoma? PMID- 15832175 TI - Expression and function of the apoptosis effector Apaf-1 in melanoma. PMID- 15832176 TI - Role of p56lck and ZAP70-mediated tyrosine phosphorylation in galectin-1-induced cell death. PMID- 15832177 TI - HIV-1 infection and AIDS: consequences for the central nervous system. AB - Infection with the human immunodeficiency virus-1 (HIV-1) can induce severe and debilitating neurological problems that include behavioral abnormalities, motor dysfunction and frank dementia. After infiltrating peripheral immune competent cells, in particular macrophages, HIV-1 provokes a neuropathological response involving all cell types in the brain. HIV-1 also incites activation of chemokine receptors, inflammatory mediators, extracellular matrix-degrading enzymes and glutamate receptor-mediated excitotoxicity, all of which can trigger numerous downstream signaling pathways and disrupt neuronal and glial function. This review will discuss recently uncovered pathologic neuroimmune and degenerative mechanisms contributing to neuronal damage induced by HIV-1 and potential approaches for development of future therapeutic intervention. PMID- 15832178 TI - To kill or be killed: how HIV exhausts the immune system. PMID- 15832179 TI - Human immunodeficiency virus type 1 (HIV-1) Vpr-regulated cell death: insights into mechanism. AB - The destruction of CD4(+) T cells and eventual induction of immunodeficiency is a hallmark of the human immunodeficiency virus type 1 infection (HIV-1). However, the mechanism of this destruction remains unresolved. Several auxiliary proteins have been proposed to play a role in this aspect of HIV pathogenesis including a 14 kDa protein named viral protein R (Vpr). Vpr has been implicated in the regulation of various cellular functions including apoptosis, cell cycle arrest, differentiation, and immune suppression. However, the mechanism(s) involved in Vpr-mediated apoptosis remains unresolved, and several proposed mechanisms for these effects are under investigation. In this review, we discuss the possibility that some of these proposed pathways might converge to modulate Vpr's behavior. Further, we also discuss caveats and future directions for investigation of the interesting biology of this HIV accessory gene. PMID- 15832180 TI - A brief history of punctoplasty: the 3-snip revisited. PMID- 15832181 TI - Preliminary results from the use of the novel Interactive binocular treatment (I BiT) system, in the treatment of strabismic and anisometropic amblyopia. AB - BACKGROUND: We have developed a novel application of adapted virtual reality (VR) technology, for the binocular treatment of amblyopia. We describe the use of the system in six children. METHODS: Subjects consisted of three conventional treatment 'failures' and three conventional treatment 'refusers', with a mean age of 6.25 years (5.42-7.75 years). Treatment consisted of watching video clips and playing interactive games with specifically designed software to allow streamed binocular image presentation. RESULTS: Initial vision in the amblyopic eye ranged from 6/12 to 6/120 and post-treatment 6/7.5 to 6/24-1. Total treatment time was a mean of 4.4 h. Five out of six children have shown an improvement in their vision (average increase of 10 letters), including those who had previously failed to comply with conventional occlusion. CONCLUSIONS: Improvements in vision were demonstrable within a short period of time, in some children after 1 h of treatment. This system is an exciting and promising application of VR technology as a new treatment for amblyopia. PMID- 15832182 TI - Modified virtual reality technology for treatment of amblyopia. AB - PURPOSE: The conventional patching/occlusion treatment for amblyopia sometimes gives disappointing results for a number of reasons: it is unpopular, prolonged, frequently resulting in poor or noncompliance, and also disrupts fusion. The aim of this research was to develop a novel virtual-reality (VR)-based display system that facilitates the treatment of amblyopia with both eyes stimulated simultaneously. METHODS: We have adopted a multidisciplinary approach, combining VR expertise with a team of ophthalmologists and orthoptists to develop the Interactive Binocular Treatment (I-BiT) system. This system incorporates adapted VR technology and specially written software providing interactive 2D and 3D games and videos to the patient via a stereo (binocular) display, and a control screen for the clinician. RESULTS: We developed a prototype research system designed for treatment of amblyopia in children. CONCLUSIONS: The result is a novel way to treat amblyopia, which allows binocular treatment. It is interactive, and as it is partially software based, can be adapted to suit the age/ability, and needs of the patient. This means that the treatment can be made captivating and enjoyable. Further research is on-going to determine the efficacy of this new modality in the treatment of amblyopia. PMID- 15832183 TI - Control of intraocular pressure after deep sclerectomy. AB - AIM: To study the long-term outcome of deep sclerectomy in patients with open angle glaucoma. METHODS: Prospective consecutive series of 43 eyes (38 patients) with medically uncontrolled open-angle glaucoma undergoing deep sclerectomy. All patients underwent clinical assessment before and after surgery at day 7 and at months 1, 3, 6, 12, 18, 24, 36. Surgical success was considered if the patient's intraocular pressure (IOP) < 22 mmHg and the IOP was lowered by more than 20% without the use of any medication. Kaplan-Meier survival curves were used to evaluate the success rate. RESULTS: The mean follow-up time was 28.1 +/- 8.2 months. Mean IOP decreased significantly from a preoperative value of 24.6 +/- 5.5 mmHg to a postoperative value of 18.5 +/- 4.6 mmHg at 36 months (P < 0.001). Microperforation of TDM occurred in three cases (7.0%) and ciliary body prolapse in one case (2.3%) but did not prevent completion of the operation. Postoperatively, hyphaema was detected in one case and shallow anterior chamber in another case and both were treated conservatively. Bleb encapsulation with elevation of IOP occurred in two cases (4.7%) and was treated with 5-fluorouracil subconjunctival injection. Goniopuncture with neodymium : YAG laser was performed in two cases (4.7%). There were no other late complications with the exception of failure of the operation. On the life-table analysis the success rate at 12, 24, and 30 months were 61.4, 36.6, and 18.9%, respectively. CONCLUSION: Deep sclerectomy reduced the IOP temporarily while minimising the risk of postoperative complications commonly encountered with standard trabeculectomy. However, after long-term follow-up surgery failed to maintain a low IOP. PMID- 15832184 TI - Differences in corneal thickness and corneal endothelium related to duration in diabetes. AB - PURPOSE: This study evaluated the differences of corneal thickness and corneal endothelial morphology in diabetes compared with age-matched, healthy control subjects; in addition, we tested for correlation according to the duration of diabetes. METHODS: Ultrasound pachymetry and noncontact specular microscopy were performed on 200 patients with diabetes and 100 control subjects. We compared the values for diabetics and normal persons with ANACOVA to adjust for age. Moreover, we examined the correlation between the subject parameters and the duration of diabetes by using a partial correlation coefficient that controlled for age. RESULTS: The diabetic subjects had thicker corneas, less cell density and hexagonality, and more irregular cell size of the corneal endothelium than did the controls (P < 0.05). Central corneal thickness and the coefficient of variation for cell size were significantly higher for diabetes of over 10 years' duration than for diabetes of under 10 years' duration (P < 0.05). The endothelial cell density and percentage of hexagonal cells were lower for diabetes of over 10 years' duration than for diabetes of under 10 years' (P > 0.05). Central corneal thickness was correlated with duration of diabetes (P < 0.05), but corneal endothelial morphology was not (P < 0.05). CONCLUSIONS: Those patients with diabetic duration of over 10 years have more corneal morphological abnormalities, especially the coefficient of variation in cell size, compared with the normal subjects. The central corneal thickness was significantly correlated with diabetic duration after controlling for age. PMID- 15832185 TI - Contamination of disposable tonometer prisms during tonometry. AB - AIM: Due to the theoretical possibility of prion transmission in applanation tonometry, many ophthalmological units in the United Kingdom now use disposable tonometer prisms. We have investigated the potential for bacterial and viral transmission from the health practitioner to the patient via disposable prisms. METHODS: All staff who perform applanation tonometry at the Sussex Eye Hospital (SEH) received a questionnaire to evaluate if the applanating face of the prism is touched during tonometry and the ease of use of the disposable prism compared to the reusable prisms that were previously used. We then cultured prisms handled by a random sample of staff members for common bacteria. Finally, we constructed a model to investigate the possibility of interpatient adenoviral transmission via disposable tonometer prisms. RESULTS: The questionnaire revealed that almost 50% of the staff admit to touching the applanating face of the tonometer prism prior to applanation. Cultures of the prisms grew a range of bacteria including Staphylococcus epidermidis, Staphylococcus aureus, and Bacillus species. The viral model suggested that adenovirus could be transmitted by applanation tonometry. CONCLUSION: The use of disposable prisms for applanation tonometry may reduce the risk of prion transmission but is not bacteriologically or virologically aseptic. This is a potential infection risk to patients. PMID- 15832186 TI - Adult ocular leech infestation. PMID- 15832187 TI - Efficacy of amblyopia therapy initiated after 9 years of age. PMID- 15832188 TI - Does lesion size determine the success rate of photodynamic therapy for age related macular degeneration? AB - Photodynamic therapy (PDT) is a new evidence-based treatment modality available for choroidal neovascularisation (CNV) secondary to age-related macular degeneration (AMD). Eligibility for PDT is based on the morphological classification of the neovascular complex, the benefit being greater in classic with no occult lesions. Lesion size is also shown to be a predictive factor for treatment benefit. This retrospective case series looked at effect of initial and final lesion size on the visual outcome of patients with subfoveal classic with no occult CNV and found that increasing initial and final lesion size is associated with poorer visual outcome. PMID- 15832189 TI - The consent and counselling of patients for cataract surgery: a prospective audit. AB - PURPOSE: The aims of ideal preoperative informed consent include educating the patient adequately to enable an autonomous decision to be made without causing undue anxiety. We study how the paternalistic and nonpaternalistic approaches meet this ideal. The influence of the new patient consent forms is also assessed. METHODS: Two cycles of a prospective clinical audit are presented. An assessment of relevant patient knowledge was performed by patient interview. Visual analogue scales were used to quantify patient anxiety. RESULTS: The first cycle, examining a paternalistic approach, demonstrated: 37% of patients understood what a cataract was and 48% understood what surgery involved. 48% misunderstood that cataract surgery was completely risk free. In total, 80% of patients undergoing second eye surgery believed that it was completely risk-free. Average anxiety visual analogue scores (VAS) for cataract surgery were low (2.89). The second cycle, examining the nonpaternalistic approach combined with the implementation of new consent forms showed that, despite more explicit repeated preoperative consent: 39% of patients understood correctly what a cataract was, 28% understood what surgery involved and 43% misunderstood that surgery was completely risk free. All patients undergoing second eye surgery thought that it was risk-free. The average anxiety VAS for cataract surgery were moderate (5.00). CONCLUSIONS: Both paternalistic and non-paternalistic approaches to informed consent are inadequate in meeting the demands of the ideal informed consent. The new patient consent forms appear to have little effect in influencing patient knowledge about their surgery. Patients undergoing second eye surgery often have an overoptimistic view of cataract surgery. PMID- 15832190 TI - Epidermal growth factor receptor expression and gene amplification in colorectal carcinoma: an immunohistochemical and chromogenic in situ hybridization study. AB - Recent data suggest that detection of epidermal growth factor receptor protein by immunohistochemistry (IHC) does not predict response to the antiepidermal growth factor receptor drug, cetuximab, in patients with colorectal carcinoma. In searching for foundation for further investigation to optimize patient selection for cetuximab therapy, this study sought to exploit the tissue microarray and chromogenic in situ hybridization techniques to evaluate the status of epidermal growth factor receptor gene amplification in colorectal cancer and its relationship with protein expression by IHC. The study included 158 primary or metastatic colorectal adenocarcinomas. Immunohistochemical results were scored as 0-3+ based on the intensity of membrane staining. The in situ hybridization signals were counted in 30 nuclei per tissue core. Overall, the rate of tissue loss was 7%, yielding 147 analyzable cases: 123 primary, 24 metastatic. Positive immunohistochemical staining of any intensity was detected in 85% (105/123) of primary and 79% (19/24) of metastatic tumors, whereas gene amplification (>5 gene copies/nucleus) was only seen in 12% (15/123) of primary and 8% (2/24) of metastatic tumors. Only 2/15 primary and 1/2 metastatic tumors that showed gene amplification were amplified at a high level (>10 gene copies/nucleus). Although a positive correlation was detected between the intensity of protein expression and the likelihood of gene amplification in both the primary (P = 0.01) and the metastatic (P = 0.05) tumors, IHC had a low specificity (17% in primary, 23% in metastatic) in predicting gene amplification. Conversely, all tumors that did not express the protein by IHC lacked gene amplification. Thus, this study shows that only a small fraction of epidermal growth factor receptor- positive colorectal carcinomas detected by IHC are associated with gene amplification. Additional studies are needed to determine whether epidermal growth factor receptor gene amplification bears any informative value in predicting response to cetuximab based therapy. PMID- 15832191 TI - Nuclear galectin-3 expression is an independent predictive factor of recurrence for adenocarcinoma and squamous cell carcinoma of the lung. AB - The tumor stage is the most powerful prognostic tool for predicting the survival rates of lung carcinoma patients. However, prognosis of individual patients is difficult in part because of the marked clinical heterogeneity among such patients. Galectins are involved in cell growth, apoptosis and cell migration features, and their diagnostic and prognostic values have already been demonstrated in various types of cancers. In the present paper we analyze the potential prognostic value of immunohistochemical galectin-3 expression in lung adenocarcinomas and squamous cell carcinomas. In all, 165 squamous cell carcinomas and 121 adenocarcinomas were immunostained for galectin-3. In each case the immunohistochemical analyses consisted of an evaluation of the percentage of tumor cells stained and the intensity of staining. An IP score (ie Intensity x Percentage) was thus determined for each lung carcinoma. A large majority of cases displayed galectin-3 expression. While the cytoplasmic staining in the squamous cell carcinomas was focal and moderately intense, the staining in the adenocarcinomas was diffuse and intense. The IP scores were significantly (P=0.0001) higher in the adenocarcinomas than in the squamous cell carcinomas. The difference in nuclear expression profiles between the two cancer types was statistically significant (P=0.0005). Cox multivariate analysis carried out on the patients' genders, the TNM classification and the galectin-3-related variables showed that of the galectin-3-related variables, only the nuclear location of galectin-3 was identified as a prognostic indicator of recurrence independent of the clinicopathological features characterizing the patients (P=0.02). The prognostic contribution of this latter variable was enhanced when the patients with relapse-free follow-ups longer than 8 months were considered (P=0.005). Galectin-3 immunohistochemical expression differs between squamous cell carcinomas and adenocarcinomas, but the nuclear expression of galectin-3 behaves as a significant prognostic predictor for all the cases as a group. PMID- 15832192 TI - Clinical utility of fluorescence in situ hybridization (FISH) in nonbrainstem glioblastomas of childhood. AB - Astrocytic gliomas are the most common pediatric brain tumors; however, nonbrainstem glioblastomas are extremely rare compared with their adult counterparts. Little information is available on the clinical significance of various molecular markers in pediatric grade IV astrocytomas. The current study was focused on the molecular analysis and clinico-pathological correlations in a set of 44 tumor samples obtained from pediatric patients with nonbrainstem glioblastomas. Fluorescence in situ hybridization (FISH) with a set of 10 commercial chromosome probes (1p36, 1q25, centromere (CEP)7, EGFR, CEP9, 9p21/p16, CEP10, 10q23/PTEN, 19p13, and 19q13) was performed. Disclosed molecular abnormalities, in descending order of frequency, included polysomy 7 (72%), loss of 10q23 (61%), loss of 9p21 (52%), loss of 1p36 (41%), gain of 1q25 (25%), polysomy 9 (16%), EGFR amplification (9%), loss of 19q13 (5%), polysomy 19 (5%), and codeletion 1p36/19q13 (2%). The overall survival time was markedly shorter only for those patients whose lesions harbored deletion of 10q23/PTEN locus (log rank test; P=0.00007). By multivariate analysis, only loss of 10q23 locus reached an independent level of prognostic value (hazard ratio=2.88; P=0.01). There were no significant differences in patient survival for other molecular abnormalities. In conclusion, a FISH analysis of 10q23 dosage should be recommended as an ancillary laboratory method that allows further clinical subdivision of pediatric glioblastomas. PMID- 15832193 TI - p16/INK4a gene methylation is a frequent finding in pulmonary MALT lymphomas at diagnosis. AB - p16/INK4a gene alterations have been associated with tumor progression in lymphoid malignancies. However, their significance in mucosa-associated lymphoid tissue (MALT) lymphoma is unclear. We investigated p16 gene methylation and mutation in a large series of untreated cases of pulmonary MALT lymphoma and diffuse large B-cell lymphoma (DLBL), and correlated p16 gene alterations with a MALT lymphoma-specific API2-MALT1 fusion and the clinicopathologic features of MALT lymphoma. The API2-MALT1 fusion was detected by multiplex reverse transcription polymerase chain reaction in 25/60 (42%) cases of MALT lymphoma, but none of 11 DLBLs. Methylation-sensitive single-strand conformation analysis showed that p16 gene methylation was frequently detected in 36/60 (60%) cases of MALT lymphoma. The gene was similarly methylated in DLBL cases (6/11, 55%). A p16 gene mutation was found in one (p16 gene-methylation) of 44 MALT lymphomas and in none of six diffuse large B-cell lymphomas. Statistical analysis showed that the p16 gene methylation status did not correlate with API2-MALT1 fusion or any of the clinicopathologic factors including serum LDH, clinical stage, and increased large cells. These findings suggest that p16 methylation is not associated with tumor progression, but may be an early event in MALT lymphomagenesis that might be maintained through the progression of the tumor. PMID- 15832195 TI - Distant metastasis in retroperitoneal dedifferentiated liposarcoma is rare and rapidly fatal: a clinicopathological study with emphasis on the low-grade myxofibrosarcoma-like pattern as an early sign of dedifferentiation. AB - The metastatic incidence of retroperitoneal dedifferentiated liposarcoma is comparatively lower than other pleomorphic sarcomas, varying widely from 1 to 18%. Low-grade dedifferentiation resembling low-grade fibrosarcoma has been recently accepted as part of the morphologic spectrum of dedifferentiated liposarcoma and was reported to have similar metastatic and survival rates to its high-grade counterpart. We sought to determine the metastatic incidence of retroperitoneal dedifferentiated liposarcoma, the clinicopathological features related to metastasis, and their postmetastatic behavior. Of all 354 retroperitoneal liposarcoma cases diagnosed at Memorial Sloan-Kettering Cancer Center during 1982-2003, we identified seven patients developing distant metastases, occurring in four females and three males, ranging from 35 to 73 years in age at presentation. They were all de novo dedifferentiated, while none of the well-differentiated liposarcoma or secondary dedifferentiated liposarcoma developed distant metastasis. Primary tumor sizes varied from 7.5 to 25 cm. All seven patients developing metastases contained >or=50% dedifferentiated elements in the primary tumor, with a predominant morphology resembling myxofibrosarcoma in five cases. The metastatic sites included the lung in four patients, somatic soft tissue in two, and liver in one. The median metastasis-free survival was 48 months, with only two patients experiencing local recurrences before developing metastasis. Six patients died of disease at median follow-up of 53 months after diagnosis and only 5 months after their first metastases. In conclusion, retroperitoneal dedifferentiated liposarcoma have a low metastatic rate, which is strongly related to de novo dedifferentiated histology that usually constitutes a prominent component of the primary tumor. Irrespective of the grade dedifferentiated liposarcoma with myxofibrosarcoma-like features should be closely monitored. Once metastases occur, they tend to follow a rapidly fatal course. PMID- 15832194 TI - Distinct progression pathways involving the dysfunction of DUSP6/MKP-3 in pancreatic intraepithelial neoplasia and intraductal papillary-mucinous neoplasms of the pancreas. AB - DUSP6/MKP-3 is identified as a candidate tumor suppressor gene for pancreatic cancer. The aim of this study was to elucidate the roles of DUSP6 in the pancreatic carcinogenesis through the pancreatic intraepithelial neoplasia and/or intraductal papillary-mucinous neoplasms, both of which are considered to be precursor lesions of invasive carcinoma of the pancreas, by comparing with involvements of other major tumor suppressive pathways. Expressions of DUSP6, CDKN2A, TP53, and SMAD4 were investigated by immunohistochemistry in a total of 206 lesions of dysplastic ductal precursors and carcinomas retrieved from 52 pancreata with invasive ductal carcinomas and 51 of those with intraductal papillary-mucinous neoplasms. The intensity of staining was evaluated in lesions at different atypical grades and statistically compared among them. Mutations of KRAS2 were analyzed by methods of the allele-specific oligonucleotide hybridization and nucleotide sequencing. In pancreata with invasive ductal carcinomas, expressions of DUSP6 were abrogated exclusively in the invasive carcinoma cells in contrast to its fairly preserved expressions in pancreatic intraepithelial neoplasia. In pancreata with intraductal papillary-mucinous neoplasms, abrogated expressions of DUSP6 were observed in a relatively small fraction of intraductal adenoma/borderlines and intraductal carcinomas. Most of the intraductal adenoma/borderline lesions with abrogation of DUSP6 harbored mutations of KRAS2. None of the molecules was associated with each other in any grade of lesions. Morphological variations of papillae of the intraductal papillary-mucinous neoplasms were evaluated and analyzed for their associations with abrogations of the molecules, which resulted in finding of no significant associations. Our results suggest that the abrogation of DUSP6 is associated exclusively with progression from pancreatic intraepithelial neoplasia to the invasive ductal carcinoma while it is potentially associated with initiation of intraductal papillary-mucinous neoplasms with mutated KRAS2, which is independent of other major tumor suppressive pathways in both types of neoplasms. PMID- 15832196 TI - Superficial vs deep pancreatic parenchymal invasion in the extrahepatic bile duct carcinomas: a significant prognostic factor. AB - Pancreatic invasion of the extrahepatic bile duct (EBD) carcinomas is known to have a poor outcome. We hypothesized that EBD carcinoma showing shallow invasion to the pancreas may have a better outcome than the usual deep pancreatic invasion. We divided 87 cases of the distal EBD carcinomas into superficial and deep pancreatic invasion groups according to degrees of the pancreatic invasion. The superficial pancreatic invasion group included cases with tumor abutting the pancreatic lobule or pancreatic parenchymal invasion equal to or less than 1 mm from the uppermost portion of the pancreatic lobule or tumors invading into the fibroadipose tissue between pancreatic lobules without parenchymal invasion. The deep invasion group consisted of tumors with more than 1 mm pancreatic parenchymal invasion. The cases with superficial pancreatic invasion showed significantly better survival rate than those with deep pancreatic invasion (P<0.001). Therefore, we recommend that a specific remark on the pathology report about the presence or absence of parenchymal invasion and the depth of invasion of the pancreas is required for managing patients and determining the prognosis. We also recommend that the current pT3 stage of distal EBD carcinomas be subdivided into superficial (pT3a) and deep pancreatic invasion (pT3b). PMID- 15832197 TI - Concordant loss of MTAP and p16/CDKN2A expression in pancreatic intraepithelial neoplasia: evidence of homozygous deletion in a noninvasive precursor lesion. AB - The p16INK4A/CDKN2A (p16) gene on chromosome 9p21 is inactivated in >90% of invasive pancreatic cancers. In 40% of pancreatic cancers the p16 gene is inactivated by homozygous deletion, in 40% by an intragenic mutation coupled with loss of the second allele, and in 10-15% by hypermethylation of the p16 gene promoter. Immunohistochemical labeling for the p16 gene product parallels gene status, but does not provide information of the mechanism of p16 gene inactivation. The methylthioadenosine phosphorylase gene (MTAP) gene also resides on chromosome 9p21, approximately 100 kb telomeric to the p16 gene. The MTAP gene is frequently contained within p16 homozygous deletions, producing concordant loss of both p16 and MTAP gene expression. Concordant loss of both p16 and MTAP protein expression can therefore be used as a surrogate marker for p16 homozygous deletion. Here we immunolabeled a series of pancreatic intraepithelial neoplasia (PanIN) lesions of various histologic grades for the p16 and MTAP gene products using a high-throughput PanIN tissue microarray (TMA) format. We demonstrate concordant loss of p16 and MTAP protein expression in 6/73 (8%) PanINs, including five high-grade lesions and one low-grade lesion. Immunolabeling for both p16 and MTAP protein expression provides a tool to evaluate tissues with intact morphology for p16 gene homozygous deletions. The concordant loss of expression of both genes in PanIN lesions demonstrates that homozygous deletions of the p16 tumor suppressor gene can occur in noninvasive precursor lesions. PMID- 15832198 TI - Choosing the greater of two goods: neural currencies for valuation and decision making. AB - To make adaptive decisions, animals must evaluate the costs and benefits of available options. The nascent field of neuroeconomics has set itself the ambitious goal of understanding the brain mechanisms that are responsible for these evaluative processes. A series of recent neurophysiological studies in monkeys has begun to address this challenge using novel methods to manipulate and measure an animal's internal valuation of competing alternatives. By emphasizing the behavioural mechanisms and neural signals that mediate decision making under conditions of uncertainty, these studies might lay the foundation for an emerging neurobiology of choice behaviour. PMID- 15832200 TI - 19th European Immunogenetics and Histocompatibility Conference, 23-26 April 2005, Istanbul, Turkey. Abstracts. PMID- 15832199 TI - WNTs in the vertebrate nervous system: from patterning to neuronal connectivity. AB - WNT signalling has a key role in early embryonic patterning through the regulation of cell fate decisions, tissue polarity and cell movements. In the nervous system, WNT signalling also regulates neuronal connectivity by controlling axon pathfinding, axon remodelling, dendrite morphogenesis and synapse formation. Studies, from invertebrates to mammals, have led to a considerable understanding of WNT signal transduction pathways. This knowledge provides a framework for the study of the mechanisms by which WNTs regulate diverse neuronal functions. Manipulation of the WNT pathways could provide new strategies for nerve regeneration and neuronal circuit modulation. PMID- 15832225 TI - Phenotype specific treatment of obstructive airways disease in infancy and childhood: new recommendations of the Swiss Paediatric Pulmonology Group. AB - In 2003 and 2004 the Swiss Paediatric Pulmonology Group (SAPP) revised the recommendations for the management of obstructive airways disease in infancy and childhood (Paediatrica 2004;15:13-28) and published recommendations for the management of acute bronchiolitis in infancy (Paediatrica 2003;14:18-21). The concept underlying these new guidelines is the fact that childhood wheezing illness encompasses a range of disorders or phenotypes with a similar clinical manifestation (wheeze, cough and breathlessness) but varying pathology, aetiology, prognosis and response to treatment. Based on the available scientific evidence, phenotype-specific and age-dependent management of wheezing illness is advocated in the revised guidelines. Major changes compared to earlier recommendations include a stepwise approach to management depending on age, phenotype and severity of disease and use of beta 2-agonists purely on an on demand basis. Comparison of these recommendations with epidemiological data on current treatment practice of obstructive airways disease in Swiss children suggests that many children with bronchiolitis and mild viral wheeze might be overtreated while management of children with severe persistent symptoms could be improved. PMID- 15832226 TI - Psychosocial and professional characteristics of burnout in Swiss primary care practitioners: a cross-sectional survey. AB - OBJECTIVE: To measure the prevalence of burnout and explore its professional and psychosocial predictors among Swiss primary care practitioners. METHODS: A cross sectional postal survey was conducted to measure burnout, work-related stressors, professional and psychosocial characteristics among a representative sample of primary care practitioners. Answers to the Maslach burnout inventory were used to categorize respondents into moderate and high degree of burnout. RESULTS: 1784 physicians responded to the survey (65% response rate) and 1755 questionnaires could be analysed. 19% of respondents had a high score for emotional exhaustion, 22% had a high score for depersonalisation/cynicism and 16% had a low score for professional accomplishment; 32% had a high score on either the emotional exhaustion or the depersonalisation/cynicism scale (moderate degree of burnout) and 4% had scores in the range of burnout in all three scales (high degree of burnout). Predictors of moderate burnout were male sex, age 45-55 years and excessive perceived stress due to global workload, health-insurance-related work, difficulties to balance professional and private life, changes in the health care system and medical care uncertainty. A high degree of burnout was associated with male sex, practicing in a rural area, and excessive perceived stress due to global workload, patient's expectations, difficulties to balance professional and private life, economic constraints in relation to the practice, medical care uncertainty and difficult relations with non-medical staff at the practice. CONCLUSION: About one third of Swiss primary care practitioners presented a moderate or a high degree of burnout, which was mainly associated with extrinsic work-related stressors. Medical doctors and politicians in charge of redesigning the health care system should address this phenomenon to maintain an efficient Swiss primary care physician workforce in the future. PMID- 15832227 TI - Antipsychotic use in patients with schizophrenia treated in private psychiatry. AB - QUESTIONS UNDER STUDY: The aim of this study was to assess antipsychotic treatment practices among private psychiatrists in Switzerland. METHODS: For each patient seen during 4 consecutive weeks, 101 participating private psychiatrists documented psychiatric diagnosis. For each schizophrenic patient, demographic details as well as treatment issues were assessed in a questionnaire-based survey. Participating psychiatrists were representative for Swiss private psychiatry with regards to gender distribution and region of practice. RESULTS: Overall, 8425 patients were assessed in our survey. Of these, 905 patients (10.7%) received a diagnosis of schizophrenic psychoses, of whom 733 details on antipsychotic treatment were documented. 73.1% of these patients received second generation antipsychotics. Most private psychiatrists prescribed antipsychotic monotherapy and maintained antipsychotic treatment according to recommendation from international guidelines. Almost half of these patients had a history of medication non-adherence, with non-compliance being the most frequent reason. CONCLUSION: The findings of this survey suggest that Swiss private psychiatrists prescribe according to international guidelines in terms of drug choice as well as maintenance treatment. Moreover they show low rates of polypharmacy and comedication practice as compared to their colleagues in other European countries. This may reflect solid experience in prescribing second-generation antipsychotics of a subgroup of private psychiatrists in Switzerland. PMID- 15832228 TI - Adherence to the Swiss guidelines for management of COPD: experience of a Swiss teaching hospital. AB - QUESTIONS UNDER STUDY: Swiss guidelines for the management of chronic obstructive pulmonary disease (COPD) were published in 2002. We aimed at assessing adherence to the proposed guidelines by the physicians in charge for all patients referred to our hospital for acute exacerbations of COPD over a one year period. METHODS: In a prospective observational study, data from a questionnaire and from records of all patients referred to our hospital with acute exacerbation of COPD were collected. Diagnostic steps as well as therapeutic and prophylactic interventions were reviewed. Where applicable, interventions were stratified according to proposed levels of evidence A-D. RESULTS: 45 patients in whom the diagnosis of COPD had been made before were included. Diagnosis was established by spirometry in 71%, in the remaining diagnosis was based on clinical grounds only. Non smoking advice was given to 69%, and 16% were offered a nicotine-replacement trial (level A). Information about a disease management plan was given in 40% of the patients (level B), 22% had done a six minute walking distance test. 27% of the patients had participated in a pulmonary rehabilitation program (level A). 93% were on regular bronchodilator therapy (level B), and 56% had regular inhaled corticosteroids (level B). CONCLUSION: Confirmation of the diagnosis of COPD by spirometry is lacking in a significant number of patients. Most patients were treated with regular bronchodilators, however, relevant over-treatment with beta adrenergic substances and overuse of inhaled corticosteroids in mild disease stages are common. Efforts for disease prevention and education as well as awareness of the potential benefits of pulmonary rehabilitation programs are still insufficient. Efforts to improve the adherence to the Swiss guidelines for the management of COPD should be intensified. PMID- 15832229 TI - Subcutaneous fat necrosis of the newborn. PMID- 15832230 TI - Improving heart rate variability (HRV) by non-pharmacological techniques. PMID- 15832231 TI - Haemodialysis catheter-related bloodstream infections: current treatment options and strategies for prevention. AB - Regardless of the repeated reservations raised by countless researchers with reference to the use of catheters as vascular access for haemodialysis (HD), central venous catheters (CVCs) remain irreplaceable tools of the modern dialysis delivery system as a reliable option for the clinical situations requiring instant access to circulation, for various reasons. Patients on long-term haemodialysis are therefore at a significantly high risk for catheterrelated bloodstream infections (CRBSI) and ensuing serious complications. Although early systemic antibiotic treatment should include the coverage for Staphylococcus aureus, the pathogen with most devastating consequences including bacterial endocarditis; optimal treatment of CRBSI while preserving the catheter site, remains contentious. Nonetheless, catheter exchange over a guide wire and antimicrobial-anticoagulant "locks" have shown promising results as novel access salvage techniques. Despite the fact that a number of novel potentially useful strategies for the prevention of CRBSI are in the pipeline; equally essential however, remains the role of rigorous implementation of standard infection control measures for hygiene and aseptic handling of CVCs in long-term HD patients. The policy of increasing the AVF (arteriovenous fistula) prevalence beyond 50% while minimising the use of CVCs, dependent largely upon the timely referrals and prudently implemented pre-ESRD program - ought to have a positive impact on long-term HD outcomes. PMID- 15832232 TI - Views of patients and physicians on follow-up visits: results from a cross sectional study in Swiss primary care. AB - PRINCIPLES: Decision making on follow-up visits is of great importance to patient physician interaction and healthcare economy. However, followup visits are a somewhat neglected topic in primary-care research, and patients' views are widely unknown. The aim was to assess the relation between the views of patients and those of physicians on follow-up visits. METHODS: We performed a cross-sectional study in six general practices in a rural area of Berne, Switzerland. Separately recorded paired data from patients and physicians on the need for and timing of revisits were analysed for differences and agreement. The revisit ultimately scheduled by the patient and the physician at the end of the consultation after sharing views on revisits was compared with the initial statements. RESULTS: A total of 250 patient-physician consultations were observed. More patients (25%) than physicians (11%) (difference 14%; 95% CI 7% to 20%) deemed a revisit unnecessary or proposed need-based revisits. Observed patient-physician overall agreement on the need or no-need of further visits was 81% (95% CI 76% to 86%). If both patients and physicians (n = 164) agreed on the need for a revisit, patients preferred slightly longer revisit intervals than did their physicians (5.9 days; 95% CI -2.6 to 14.4). The revisit interval recorded in the appointment calendar at the end of the consultation nearly always matched the patient's preferences. CONCLUSION: In one fifth of all consultations, patients' views on revisits differ from those of their physicians, with patients choosing a more liberal involvement in follow-up visits. Actively involving patients in the process of revisit handling might have an influence on revisit frequencies. The impact of the shared revisit handling on patientrelevant outcome measures and healthcare economy needs further consideration.. PMID- 15832233 TI - Pitfalls in the emergency department triage of frail elderly patients without specific complaints. AB - QUESTION UNDER STUDY: Elderly patients represent an increasing proportion of emergency department (ED) admissions. When no specific complaint is identified, the reason for referral is commonly called "home care impossible". The aim of this study was to describe a population of elderly patients who present to the ED of a 1200-bed university hospital without specific complaint, and to assess how they were evaluated in the ED. METHODS: Data on triage, mode of admission and discharge were collected. After the initial evaluation in the ED, patients were classified in two categories: (1) patients identified with a medical problem requiring rapid care or investigation, (2) patients without a medical problem considered as true "home care impossible". These latter patients underwent a complete assessment using the Minimal Data Set-Home Care (MDS-HC). RESULTS: During the 10-week study period 253 patients (mean age 81 years) were referred because of "home care impossible". An acute medical problem was identified in 129 of those patients (51%). All these patients were triaged in lower acuity categories. 33 (26%) were undertriaged due to (1) absence of vital signs measurement, (2) poor recognition of neurological symptoms, (3) atypical clinical presentation. The remaining patients were considered as true "home care impossible". The MDS-HC evaluation revealed a high level of biopsychosocial comorbidities. CONCLUSIONS: Frail elderly patients admitted without specific complaints are at risk of inappropriate or delayed evaluation due to undertriage at the door of the ED. A more specific geriatric assessment should be integrated early in the triage process of these patients. PMID- 15832234 TI - Impact of aspirate smears and trephine biopsies in routine bone marrow diagnostics: a comparative study of 141 cases. AB - The diagnostic impact of bone marrow cytology in combination with flow cytometry analysis of aspirate smears and bone marrow histology together with immunohistochemical examination of trephine biopsies was compared in 141 routine cases. Diagnoses achieved by the two methods were concordant in 80.5% of cases. In discordant cases, clinical follow-up data of at least one year confirmed the correctness of cytological and histological diagnoses. For infiltration by malignant disease, both methods were concordant in 86.5% of samples and correlated well for the degree of infiltration (r = 0.64, p <0.001). Overall, regression analysis showed a good correlation for cellularity (r = 0.67) lymphopoiesis (r = 0.75), granulopoiesis (r = 0.73) and megakaryopoiesis (r = 0.65) while erythropoiesis displayed a lower degree of correlation (r = 0.43, all p <0.001). Regression analysis on all immunological data obtained by flow cytometry (FC) and immunohistochemistry (IHC) showed a good overall linear correlation (r = 0.67, p <0.001), but significant differences were found for a few phenotypic markers. Furthermore, the correlation was found to be dependent on IgG subclasses and the fluorochromes used for FC. Thus, analyses with IgG2 antibodies and phycoerythrin (PE) as fluorochrome showed significantly more expression than IHC. In conclusion, cytology and histology, both in association with the respective immunophenotyping, are of equal value in bone marrow diagnostics and should be used in combination. However, in some specific settings, one of the two procedures might be preferable. PMID- 15832235 TI - Bilateral black eyes, "raccoon's eyes", resulting from plucking of eyebrows. PMID- 15832236 TI - [Gender-medicine -- terra incognita in anaesthesiology?]. PMID- 15832237 TI - [Sex/Gender research -- introduction and anesthesiological aspects]. AB - On the one hand gender studies are based on the differentiation of biological (sex) and social (gender) identities. Above all, gender studies in public health research are seen as an biological, psychological and social entity. To understand the complexity linked to the term "gender" in medicine, different perspectives are required. Apart from the uncertainty, which may occur by the changing perspective on gender, it also holds the option for innovation in anesthesiology, too. The goal of gender based medicine is an optimized, individual adjusted medical care. Precondition to realize this aim is the understanding of the genesis, the consolidation, and the elimination of gender biases in all fields of research, publications and education. The transfer of methods and guidelines for gender sensitive research, adapted from the public health research, allows an access to detect the meaning of gender specifics and gender differences, to verify its clinical relevance and to exercise the results in clinical daily work. With gender sensitive methods desiderata should be reduced, and methodological standards for gender sensitive medical/clinical research can be developed and implemented into medical education. In the light of knowledge about sex specific differences in pharmacokinetic and pharmacodynamic as well as undesired drug interactions, basic research may help to overcome the gender gap and to individualize and optimize the anesthesiological approach as well. A main problem in implementing gender studies into medicine is the limitation of sex sensitive biomedical significance. Nevertheless, the already existing measures in public health should be evaluated and modified for clinical practice. The current review is to be read as an introduction into different aspects of gender studies in medicine, in order to extend the understanding of its importance for anesthesiology, too. It shows that the relevance transcends still existing studies based on biomedical gender diversities. PMID- 15832238 TI - [Extracorporeal blood purification in severe liver failure with the albumin dialysis MARS -- impact on relevant intensive care parameters]. AB - Extracorporeal liver support methods have been tested for over 50 years now. Standard techniques of blood purification like dialysis, adsorption, hemo- and plasma filtration as well as bioreactor-based approaches using liver cells or tissues have been used. Most clinical experience, however, is limited to use in acute liver failure (ALF). Since 1993, the Molecular Adsorbent Recirculating System (MARS) has been used clinically -- a system that combines dialysis, filtration and adsorption in a biocompatible method. Human serum albumin (HSA) acts as a selective molecular adsorbent binding protein-bound compounds like bile acids or bilirubin. These substances can contribute to the maintenance or even further aggravation of liver failure. They are linked with the pathogenesis of hyperdynamic hypotonic circulation, hepatic encephalopathy, hepatorenal syndrome, impaired hepatic protein synthesis, and intractable pruritus seen in chronic liver failure. HSA takes over the toxic substances from a patient's blood and passes through a remote detoxification process including bicarbonate-dialysis and a two-step adsorption. It is then recirculated in the patient's blood. Up to today, more than 4000 patients have been treated in approximately 16,000 single sessions. Thus, MARS represents the most frequently used liver support method at the present time. In addition to ALF, mainly acute decompensations of chronic liver failures (ACLF) have been treated. The impact of the extracorporeal treatment on relevant medical parameters of intensive care medicine is discussed with regard to the specific situation of the liver-failure patient (susceptibility to infection, atypical picture and course of infection, coagulation disorders and bleeding tendencies). PMID- 15832239 TI - [Acute treatment of stable hemodynamically tolerable ventricular tachycardia]. AB - Sustained monomorphic ventricular tachycardia is a potentially life-threatening ventricular tachycardia. The arrhythmia can be treated with antiarrhythmic drugs, when the patient is hemodynamically stable. Class IC antiarrhythmic drugs like flecainide and propafenon or ajmaline (class IA/IC) are very effective. The use of lidocaine is obsolete. Sotalol is also effective, however, caution is advised because of its significant beta-sympatholytic properties. Amiodaron is frequently used, although it has never been studied in the setting of stable ventricular tachycardia. Earlier studies have demonstrated that it is effective in the treatment of recurrent, hemodynamically unstable ventricular tachycardia. When a particular drug does not terminate tachycardia, further drugs should not be given. Electrical cardioversion is preferred. PMID- 15832240 TI - [Effects of sevofluran on cell division and levels of sister chromatid exchange]. AB - OBJECTIVE: Purpose of the study was to investigate the mitotic index (MI) and sister chromatid exchange (SCE) levels to identify the mutagenic and carcinogenic effects of sevoflurane (sevoflurane). METHODS: 42 non-smoking male and female turkish patients of ASA-risk I and II were included. The patients received an anaesthesia induction with 8 % sevoflurane in 100 % oxygen ("tidal volume methode") and 0.1 mg/kg BW vecuronium for neuromuscular block and endotracheal intubation. Anaesthesia was maintained with 2.0 - 2.5 sevoflurane in 60 % N(2)O and 40 % O(2). Four 5 ml venous blood samples werde taken: before induction (control), 60 minutes, 24 hours and 5 days after sevoflurane anesthesia. Samples were prepared according to the periferic blood culture assay, modified by Morhead and co-workers, and levels of MI and SCE were examined. RESULTS: 60 minutes after sevoflurane-anaesthesia a significant decrease of MI was found compared to controls (p < 0.01). This depression was lower after 24 hours (p < 0.05) and reversible after 5 days. SCE increased significantly during 60 minutes of anaesthesia (p < 0.001), was also lower after 24 hours (5.6 +/- 2.4 vs. 4.4 +/- 1.7) and returned to normal levels after 5 days (p > 0.05). CONCLUSION: The application of sevoflurane for anaesthesia may influence the cell division in humans and may have a mutagenic effect on DNA at the cell level, which is reversible. PMID- 15832241 TI - [Effect of rocuronium on the diaphragm, musculus adductor pollicis and orbicularis oculi in two groups of different age]. AB - OBJECTIVE: Onset time and recovery from non depolarising neuromuscular blockade depends on the tested muscle and is influenced by the age of the patient. This study compares the neuromuscular blocking effect of rocuronium on the diaphragm, adductor pollicis and orbicularis oculi muscle in young and elderly patients. METHODS: After institutional ethics committee approval and written informed consent, 20 adult patients (ASA I - II), age 18 - 59 and > 65, have been included. Neuromuscular response was measured by accelerography for the adductor pollicis and orbicularis oculi muscle. Monitoring of the diaphragm consisted of measurement of the airway pressure against an occluded tracheal tube during magnetic phrenic nerve stimulation. Onset time and recovery were measured after injection of 0.6 mg/kg Rocuronium. RESULTS: The adductor pollicis had the fastest onset time ( young 2.3 min, old 2.2 min), followed by diaphragm ( young 3.6 min, old 3.4 min) and orbicularis oculi muscle ( young 3.7 min, old 4.8 min). There was a complete blockade of the diaphragm in 50 % of all patients (Adductor pollicis 100 %, orbicularis oculi 40 %). Neuromuscular recovery, recovery index and TOF 0.8 differed significantly between young and elderly patients. Onset of recovery was earlier at the diaphragm ( young 15.9 min, old 22.0 min) compared to the peripheral muscles (adductor pollicis young 25.6 min, old 37.9 min, orbicularis oculi young 23.8 min, old 27.5 min). CONCLUSION: 2 fould ED95 of rocuronium often results in an incomplete neuromuscular blockade of the diaphragm. Therefore monitoring of the peripheral muscles in patients given a single dose of rocuronium often overestimates the degree of diaphragmatic relaxation, but is a save predictor of recovery. Especially in elderly patients were prolonged neuromuscular blockade should be expected, a neuromuscular monitoring is recommended. PMID- 15832242 TI - [Rigor mortis -- a definite sign of death?]. AB - In the past years an ongoing controversial debate exists in Germany, regarding quality of the coroner's inquest and declaration of death by physicians. We report the case of a 90-year old female, who was found after an unknown time following a suicide attempt with benzodiazepine. The examination of the patient showed livores (mortis?) on the left forearm and left lower leg. Moreover, rigor (mortis?) of the left arm was apparent which prevented arm flexion and extension. The hypothermic patient with insufficient respiration was intubated and mechanically ventilated. Chest compressions were not performed, because central pulses were (hardly) palpable and a sinus bradycardia 45/min (AV-block 2 degrees and sole premature ventricular complexes) was present. After placement of an intravenous line (17 G, external jugular vein) the hemodynamic situation was stabilized with intermittent boli of epinephrine and with sodium bicarbonate. With improved circulation livores and rigor disappeared. In the present case a minimal central circulation was noted, which could be stabilized, despite the presence of certain signs of death ( livores and rigor mortis). Considering the finding of an abrogated peripheral perfusion (livores), we postulate a centripetal collapse of glycogen and ATP supply in the patients left arm (rigor), which was restored after resuscitation and reperfusion. Thus, it appears that livores and rigor are not sensitive enough to exclude a vita minima, in particular in hypothermic patients with intoxications. Consequently a careful ABC check should be performed even in the presence of apparently certain signs of death, to avoid underdiagnosing a vita minima. Additional ECG- monitoring is required to reduce the rate of false positive declarations of death. To what extent basic life support by paramedics should commence when rigor and livores are present until physician DNR order, deserves further discussion. PMID- 15832243 TI - [Perioperative treatment of patients with Diabetes mellitus]. PMID- 15832246 TI - [Tactile-kinesthetic responsiveness in children with cleft lip palate]. AB - BACKGROUND: Literature provides ample evidence that tactile-kinesthetic responsiveness is of basic importance for a normal speech development. The purpose of this study was to examine tactile-kinesthetic functions in children with cleft lip palate as they are suspected to be at risk to develop disturbances in this respect because of their anatomic defect. MATERIALS AND METHODS: Tactile kinesthetic responsiveness was measured using the specific DEF-TK questionnaire as previously proposed by Kiese-Himmel. Twenty-seven children aged 1;3 to 7;11 years with cleft lip palate were examined. Forty-two children with an isolated speech development disorder served as age matched controls. RESULTS: The results of both the study and the control group did not differ on a statistically significant level (p > 0.05). Compared to the data taken from the test manual both groups exhibited a significant number of questionnaires which could not be analyzed because of too many unobservable (not missing!) items, yet, this number did not differ between the two groups. So a "best case" and a "worst case" were calculated but did not reveal significant differences either. CONCLUSION: Surprisingly and in contrast to the results reported in the literature the DEF-TK questionnaire exhibited methodical shortcomings with a significant number of cases in which it could not be analyzed due to too many unobservable items. So, the results obtained can only be discussed under reserve: They indicate that children with cleft lip palate do not exhibit more tactile-kinesthetic problems than children with an isolated speech development disorder and thus do not require specific therapy in this field. PMID- 15832247 TI - [Endolaryngeal distance measurement by laser light projection--from the idea to clinical application]. AB - INTRODUCTION: In clinical and scientific examinations with rigid and flexible endoscopes quantitative morphometric measurements of intralaryngeal structures have been difficult whereas qualitative laryngoscopic examinations are routine procedures. MATERIAL AND METHODS: The development of a two-point light projection method for endolaryngeal morphometry in its different phases is described. The way from a first prototype for endolaryngeal morphometry that works with a prism technique to the latest, most modern prototype, which works with a double reflecting mirror is shown. RESULTS: In combination with a newly developed PC software the latest prototype for two-point laser-light projection enables an accuracy of 95 % to 99 % in the measurement of intralaryngeal distances. The contrast of the laser beams to the mucous membrane of the larynx could be optimized by the use of green--instead of red--laser light. CONCLUSION: Morphological measurements within the larynx are now possible for clinical and scientific questions. Former limitations in quantitative morphometric examinations will be reduced. PMID- 15832249 TI - [Prevention of recurrent laryngeal nerve paralysis by demonstration of the nerve during thyroid surgery]. AB - INTRODUCTION: Recurrent laryngeal nerve paralysis is one of the major complications of thyroid surgery. The importance of dissection and exploration of the recurrent laryngeal nerve during thyroid surgery remains controversial. METHODS: 74 thyroid gland operations with obligatory dissection and exploration of the recurrent laryngeal nerve were analysed. RESULTS: 118 recurrent laryngeal nerves were explorated in these operations. Transitory postoperative vocal cord paralysis was observed in 4 patients. In one patient vocal cord paralysis was permanent. This results in a transitory vocal cord paralysis rate of 3.4 % and a permanent vocal cord paralysis rate of 0.85 %. DISCUSSION: Our results and the literature review indicate that dissection and visualization of the recurrent laryngeal nerve can reduce the risk of permanent paralysis to a minimum. Obviously mechanical trauma like compression and crushing due to dissection do not increase incidence of permanent nerval disorders. Certain dissection and identification of the recurrent laryngeal nerve during thyroid surgery is recommended in principle. PMID- 15832248 TI - [Voice function and quality of life--patients with laryngeal carcinoma following minimal-invasive laser surgery and function voice rehabilitation]. AB - BACKGROUND: Quality of life is a relevant criterion for therapeutical outcome in laryngeal carcinoma patients. The aim of this study was to assess the self related complaints in quality of life and functional outcome of voice among persons who had undergone laryngeal cancer surgery. METHODS: The examination took place 67 months (SD 34.8) after the last surgery, on average and finishing of 60 therapy sessions (median). First a measurement with the Goettinger Hoarseness Diagram (GHD) was carried out. Next the subjects answered self-reported standardized questionnaires for life quality in oncological patients on the same day: EORTC-QLQ-C30 and 3 scales of the German Fragebogen zur Lebenszufriedenheit (health-related fulfillment, fulfillment with work/occupation, fulfillment with social environment). For disease-related symptoms of different head and neck domains the EORTC-HandN35 was administered. SUBJECTS: Forty-four patients (37 males, 7 females) who were treated primarily with minimal invasive laser surgery followed by functional voice rehabilitation (Goettinger Konzept). Mean age: 60.1 (SD 9.7) years. RESULTS: The acoustic voice quality according to the GHD showed a significant correlation with self-reported "Somatic Functioning" (- 0.32, p = 0.04) and "Social Functioning" (- 0.38, p = 0.01) in the EORTC-QLQ-C30 as well as with the symptom scales "Speech" (0.45, p < 0.0024) resp. "Social Contact" (0.45, p = 0.0021) in the EORTC-HandN35. Also hoarseness and health-related fulfillment (- 0.36, p = 0.02) as well as fulfillment with work/occupation (- 0.33, p = 0.03) correlated significantly. On the other hand, there was no significant association between the GHD and the assessment of global quality of life. CONCLUSION: Postoperative phonatory reductions in patients with laser surgically treated laryngeal carcinoma have only a slight influence on overall quality of life. The same is true for the GHD. Assessment of self-reported quality of life in recommendation of oncologic treatment may not be able to replace the objective findings, but may be useful to detect other problems, e.g. psychosocial. PMID- 15832250 TI - [Is the observation of patients with sleep-apnea-syndrome after surgery of the upper airway in an intensive care unit generally necessary?]. AB - BACKGROUND: Although it is known that after surgery of the nose and/or the paranasal sinuses serious complications can arise for patients suffering from Sleep-Apnea-Syndrome (SAS), there exists no general recommendation for postoperative care of these patients. This retrospective analysis is dealing with the question whether it is generally necessary to observe SAS-patients after nasal surgery including intubation in an Intensive Care Unit (ICU). PATIENTS AND METHODS: 24 Patients of the ORL-Dept., Marienkrankenhaus Hamburg, suffering from SAS underwent surgery of the nose, the paranasal sinuses and/or the pharynx including total intravenous anesthesia (TIVA) during the period of 1. 10. 2000 until 1. 5. 2004. SAS was diagnosed in 6 cases due to defined clinical criteria and in 18 cases due to the polysomnographic findings in the sleeping laboratory's examination. All patients were observed postoperatively for one night in an ICU. The anesthesia protocol and the intensive care curve of each patient were systematically evaluated with special regard of the following parameters: Risk factors (Body Mass Index; other diseases, ASA-classification), premedication drugs, duration of the surgery, drugs for pain relief, lowest O2-saturation of blood, lowest heartrate, highest systolic blood pressure, adverse effects, intensive care interventions. RESULTS: Intensive care interventions were never needed. 2 patients received a low dosage of oxygeninsufflation via a face mask, in 5 cases calcium-antagonist drugs were administered due to high blood pressure and in 1 case Metamizole administration was necessary due to high temperatures. An accompanying bradycardia of the same patient was treated by administration of Atropine. The lower average O2-saturation was 93.6 +/- 1.7 % (Minimum value: 89 %). The maximum systolic blood pressure was 165.8 +/- 21.2 mm Hg and the lowest average heart rate was 65.4 +/- 13.2 bpm. CONCLUSIONS: Patients suffering from a mild to moderate SAS do not need a general postoperative surveillance in an ICU if the chosen form of anesthesia is considered concerning this sickness. PMID- 15832251 TI - [Transmastoidal resection of an endaural encephalocele--a case report]. AB - BACKGROUND: There are transmastoid, transtemporal and combined approaches for surgical management of endaural encephaloceles. The advantages of a transmastoid approach are the exploration and option of surgical treatment of the middle ear cavity without craniotomy. Nevertheless there are recent studies preferring the transtemporal or combined approach because of the better survey. PATIENT: We present a 27-year old male patient, who developed hearing loss and otorrhea of the right side. He underwent middle ear surgery associated with cholesteatoma three times before. On examination there was a smooth pulsatile mass taking origin in the roof and almost filling the external auditory canal. High resolution CT and MRI scans of the petrous bone demonstrated a spherical soft tissue mass communicating with the middle cranial fossa. The encephalocele was exposed and resected through a transmastiodal approach. Behind the cele a relapsing cholesteatoma was found and resected in the same session. The laterobasilar defect was sealed with ear cartilage and a perichondrium flap. CONCLUSION: There are three different surgical approaches to manage laterobasilar encephaloceles. Due to the history of a previous cholesteatoma we performed a transmastoidal approach. A transtemporal approach alone would have failed to notice the relapsing cholesteatoma and would have led to a greater trauma. PMID- 15832252 TI - [Up to date: therapy of sudden hearing loss]. AB - Sudden sensorineural hearing loss is thought to be of different origin. Disturbances of microcirculation, autoimmune pathology and viral infection are among the most likely causes. The shape of the puretone audiogram can be a useful clue to its origin. Following is a description of five different forms of sudden hearing loss, its suspected pathogenesis and different therapeutic strategies recommended for the treatment of sudden hearing loss. PMID- 15832255 TI - [Acknowledgement of a laryngeal carcinoma after exposure to ionising radiation as an occupational disease]. PMID- 15832257 TI - [Review and categorization of quinolone antibiotics]. AB - No standard categorization of quinolone antibiotics into generations may be found in either Czech or world literature. The author recommends a categorization into four groups defined according to their spectrum of action and utilization: 1) preparations for the treatment of urinary tract infections; 2) systemically acting quinolones chiefly efficacious against Gram-negative bacteria; 3) so called respiratory quinolones; and 4) quinolones with a very broad spectrum of action suitable for the treatment of very complicated infections. The author describes the chief characteristics of the most important quinolone antibiotics, including preparations either in their development stage or whose development has been prematurely interrupted because of adverse side-effects. The list includes all preparations that are or were temporarily registered in the Czech Republic. PMID- 15832258 TI - [Crohn's disease and infectious agents]. AB - The author discusses possible etiological causes of Crohn's disease taking into consideration the undoubtedly significant impact of the constitution of the intestinal microbial flora. The author pays special attention to Yersinia; their presence in the intestines is believed to be the chief mechanism of subsequent immunity events resulting in the development of the disease. The author briefly discusses the possibilities of differential diagnosis and of treatment. He mentions the profitable cooperation of infectiousdiseases specialists and gastroenterologists in the diagnosis of diarrhoeal disorders. PMID- 15832259 TI - [Molecular biology characteristics of ESBL-positive strains of Klebsiella pneumoniae collected in the Neonatal Unit of the Teaching Hospital in Olomouc]. AB - BACKGROUND: One of the problems of contemporary medicine is an increasing number of bacterial strains with hazardous phenotypes of resistance. This is also true for neonatal units where nosocomial infections caused by multiresistant bacteria pose a serious threat to newborns. The feared bacterial pathogens include Klebsiella pneumoniae strains producing AmpA Extended-Spectrum Beta-Lactamases. The study focused on the molecular biology characteristics of ESBL-positive strains of K. pneumoniae collected in the Neonatal Unit of the Teaching Hospital in Olomouc (THO). MATERIALS AND METHODS: Clinical material from newborns hospitalized in the THO Neonatal Unit between January and June 2004 was used to isolate and determine K. pneumoniae strains by standard identification procedures. Their susceptibility to antibiotics was tested using a dilution micromethod. A Double-Disk Synergy Test was used for phenotype determination of ESBL production. The bla gene coding ESBL production was demonstrated by PCR. Molecular biology characteristics of ESBL-positive strains utilized the genomic DNA isolation, XbaI restrictase digestion and PFGE differentiation. The acquired restriction maps of individual isolates were compared using the GelCompare software and their relationship was determined. The selection pressure of antimicrobial agents was assessed according to the absolute number of defined daily doses of individual antibiotics. RESULTS: During the monitored period, 112 K. pneumoniae strains were isolated in total. In 22 of them (19.6%), the TEM-type ESBL production was determined. ESBL-positive strains were only observed in upper respiratory tract and rectal swabs collected from newborns with no signs of infection. The molecular biology analysis showed that 21 ESBL-positive strains had an identical restriction profile, i.e. they were very likely to be identical. The selection pressure of third- and fourth-generation cephalosporins was very low over the observed period and their consumption accounted for 1.9 % of all administered antimicrobial agents. CONCLUSION: The results presented above suggest that ESBL-positive strains of K. pneumoniae occurred in the THO Neonatal Unit due to clonal and horizontal spread from an unidentified source. PMID- 15832260 TI - [Present possibilities of anti-meningococcal vaccination]. AB - In the form of answers to the most frequent questions the author reviews the present development of an antimeningococcal vaccine and describes the existing possibilities of vaccination. The article broaches the history of vaccine development, categories of antimeningococcal vaccines, their weaknesses and advantages. The author discusses the issue of a vaccine against meningococcus B, the difference between a polysaccharide and conjugated vaccine, the vaccination strategy in the Czech Republic and in the world and briefly describes the antimeningococcal immunity of our population. PMID- 15832264 TI - Differences in oestrogen and progesterone receptors, HER-2, p53 expression and proliferation in ductal breast cancers in relation to histopathological grade. AB - In case of breast cancer the grade of differentiation and expression of oestrogen and progesterone receptors falls within the first category of prognostic factors according to the College of American Pathologists. HER-2, p53 and Ki67 belong to the second category and their significance still awaits confirmation. The aim of the present study was to examine the relationship between the intensity of expression of oestrogen receptors (ER), progesterone receptors (PgR), HER-2, p53 and Ki67 in cells of ductal breast cancer of G1, G2 or G3 differentiation grade. In paraffin sections of 60 ductal breast cancers (20 cases in G1, 20 in G2 and 20 in G3), immunocytochemical reactions were performed to detect the expression of ER, PgR, HER-2, p53 and Ki67. Following a semi-quantitative appraisal of the preparations under examination, appropriate statistical tests were used to document significant relationships. We noted significant positive correlations between ER and PgR (the entire group studied, G1-3, and the G1 group), HER-2 and p53 (G2) and between p53 and Ki67 expression (G2). Significant negative correlations were found between ER and p53 (G1-3), PgR and p53 (G1-3, G1, G3) and between PgR and Ki67 (G1-3, G2). The studies performed demonstrated distinct relationships between the expression intensity of various proteins in tumour cells in relation to the grade of differentiation of the tumour. We also showed that a parallel determination of ER, PgR and p53 expression may carry high predictive value as to response to tamoxifen treatment. PMID- 15832263 TI - Morphological studies in modern teratological investigations. AB - Despite the variety of modern molecular techniques available, examination of foetal anatomy is still a fundamental part of teratological studies in evaluating the developmental toxicity of xenobiotics or other non-chemical factors. The article presents contemporary methods of embryotoxicity and foetotoxicity assessment. A single alizarin red S and double alcian blue followed by alizarin red S staining, as well as various methods of soft tissue examination are discussed. PMID- 15832265 TI - Compensating crural anastomoses in chronic critical limb ischaemia. AB - Compensating crural anastomoses develop in patients with multi-level occlusion of the calf arteries in the course of atherosclerosis, arteriitis, diabetes, and in vascular malformations of the limbs. The peroneal artery is frequently the only patent calf vessel, especially in diabetic patients who have advanced tibial occlusive disease. The purpose of this study was to identify different types of compensating crural anastomoses in chronic critical limb ischaemia. Using combined anatomical-radiographic and statistical methods, 86 compensating crural anastomoses were studied in 59 specimens of lower limbs (amputated at the thigh) in the course of chronic critical ischaemia. Three types of compensating crural anastomosis and their components were identified. The most common type (55.8%) was the posterior tibioperoneal anastomosis. Less common (23.3%) was the intertibial anastomosis and least common (20.9%) the anterior tibioperoneal anastomosis. The posterior tibioperoneal anastomosis was concurrent with anterior tibioperoneal anastomosis in 26.3% of cases and with the intertibial anastomosis in 15.3% of cases. The great importance of the peroneal artery in the formation of natural crural collateral circulation should encourage vascular surgeons to consider peroneal bypasses. PMID- 15832266 TI - The development of Meckel's cartilage in staged human embryos during the 5th week. AB - The study was conducted on 15 embryos aged 5 weeks. The primordium of Meckel's cartilage appears at stage 13 (32 days) as a rounded structure composed of fusiform and polygonal cells, which blend with other cells of the mandibular process. At stages 14 and 15 (33 and 36 days) Meckel's cartilage forms a well delineated core of small densely packed cells. PMID- 15832267 TI - Morphometry of the pancreas in human foetuses. AB - With the use of conventional anatomical dissection, radiography, digital and statistical analysis, morphometry and skeletopy of the pancreas was carried out in 60 human foetuses of both sexes (28 female, 32 male) between the 17th and 40th week of intrauterine life. The material was fixed in a 10% formalin solution. The age of the foetuses was determined by crown-rump (CR) length measurement on the basis of the Iffy et al. tables. Photographic documentation was made and then digitally processed in the Computer Image Digital Analysis System. The following parameters were taken into account: the length and width of 3 parts of the pancreas, namely the head, corpus and tail. Additionally, radiograms were made to obtain a projection of the gland on the vertebral column. Development of the pancreas was correlated with the age of the foetuses calculated on the basis of crown-rump (CR) length measurements. The correlation coefficient with CR was 0.998 for the pancreas length, 0.709 for the width of the head, 0.703 for the width of the corpus and 0.712 for the width of tail. Gender dimorphism was not found (p > 0.05) with regard to the morphometry of the pancreas. In the material under examination the pancreas did not change its position in relation to the vertebral column. The head projected on the vertebral column in the range Th(12) L(2) (most frequently L(1)-L(2)), the corpus on Th(12)-L(2) and the tail on Th(11). PMID- 15832268 TI - Analysis of calretinin immunoreactivity in the rat piriform cortex after open field stress during postnatal maturation. AB - In our study we used c-Fos protein to identify whether cells containing calretinin (CR) in the rat piriform cortex are engaged in the response to stress stimulation and to find out how this expression changes during maturation (PC). The material consisted of Wistar strain rats of between 0 and 120 days of age divided into 9 groups. Each group consisted of 5 experimental and 3 control rats. Animals from the experimental groups were exposed to the open field test throughout 10 minutes. The control animals were kept in a home cage. In all age related control rats weak c-Fos immunoreactivity was observed. Our results showed that cells containing c-Fos following an acute open field test were observed predominantly in layers II and III of the PC just after birth. Their number then increased and stabilised on P30. We had already observed immature CR-ir cells at birth. In the 4th week of life these neurons achieved maturity. Their number increased to P90 and decreased in older animals. CR-ir neurons were localised mainly in layer II and to a lesser degree in layers III and I of the PC. Double immunostaining c-Fos/CR revealed that the level of co-localisation was low. Only small differences were observed between the anterior and posterior parts of the PC. In the anterior part a higher number of CR-ir neurons was found. The difference in the level of co-localisation between the anterior and posterior parts was age-related and differentiated. Our results may suggest that during maturation CR-ir neurons of the piriform cortex are not the main population engaged in response to the open field test. PMID- 15832269 TI - Histological and histochemical studies on the olfactory rosette of Mugil parsia (Hamilton). AB - The structure and functions of the olfactory organs in Mugil parsia (Ham.) has been described. Histologically each lamella consists of supporting, olfactory receptor, basal, labyrinth and mast cells. The distribution and localization of acid and neutral mucins in the various cells of olfactory epithelium in M. parsia has been studied histochemically. Variations in the localization of glycogen in the different cells of the olfactory epithelium have been correlated with the functional significance of the region concerned in the fish studied. PMID- 15832270 TI - A new variant of aberrant left brachiocephalic trunk in mam: case report and literature review. AB - Importance is placed on aberrant arteries in the radiological and surgical literature. A normal left brachiocephalic trunk is characteristic for the right aortic arch. However, an aberrant left brachiocephalic trunk arising as the last branch of the aortic arch on the left side has not yet been described in the literature. Described here is a new variant of the retro-oesophageal aberrant left brachiocephalic trunk, occasionally observed in a patient during diagnostic investigation or surgical treatment for steno-obstructive involvement of the carotid district. The triple anomaly of the left aortic arch consisted of: 1. the presence of a hypoplastic left brachiocephalic trunk behind the oesophagus, 2. the absence of a brachiocephalic trunk on the right side and 3. separate origins of the arteries on the right side, with the right common artery preceding the right subclavian artery. In front of the trachea an 8-mm prosthetic PTFE was implanted from the proximal segment of the right subclavian artery to the junction of the left common carotid and left subclavian arteries. The author demonstrates the inadequacy of auxiliary investigations to detect aberrant arteries, which may only be identified precisely intra-operatively. PMID- 15832272 TI - Two cases of inferior vena cava duplication with their CT findings and a review of the literature. AB - Duplication of the inferior vena cava (IVC) is the most common anomaly to affect the vena cava. Variations in the IVC are diagnosed in routine dissection studies, in retroperitoneal surgeries, or in computerised tomography (CT) sections ordered for various reasons. In this paper we present two cases of a double IVC together with the CT findings. The duplication might have occurred during embryological development. Although venous anomalies are rare, they have particular importance with respect to the interruptions that may occur during retroperitoneal and thoracic surgery or in the treatment of thrombo-embolic disease. PMID- 15832271 TI - Abnormal location of papilla of Vater: a cadaveric study. AB - We report a case of a male cadaver aged 72 years with an ectopic location of the papilla of Vater. The ectopic papilla was situated at the supero-posterior border of the 3rd portion of the duodenum at a distance of 0.9 cm from the limit of the 2nd and 3rd portions of the duodenum. The frequency of this anomaly fluctuates between 0 and 11.83% and when the papilla is located distal to its usual position the usual location is in the proximal 2 cm of the 3rd part of the duodenum. We refer to the possible difference in the papilla's location between patients and cadavers and call attention to the differential diagnosis with spontaneous or surgical fistulae. PMID- 15832273 TI - Bilateral alar thoracic artery. AB - During a routine dissection a superficial artery was observed coursing subcutaneously at the anterior border of the axillary base towards the thoracic wall and bilaterally at the lower border of the pectoralis major muscle. On the right side it originated from the 3rd part of the axillary artery but on the opposite side the origin was from the first centimetre of a left radial artery originating directly from the axillary artery together with the left brachial artery. Apart from the bilateral absence of the deep brachial artery, no other anomalies were identified at this level. This variant corresponds to the alar thoracic artery, an unusual and rarely reported artery. The literature on the subject contains no reference either to the bilateral evidence for the alar thoracic artery or to the possibility of an origin from a high radial artery. The presence of such an alar thoracic artery may interfere with surgical access within the axillary fossa and should be taken into consideration. PMID- 15832287 TI - Novel tumor necrosis factor-knockout mice that lack Peyer's patches. AB - We generated a novel tumor necrosis factor (TNF) null mutation using Cre-loxP technology. Mice homozygous for this mutation differ from their "conventional" counterparts; in particular, they completely lack Peyer's patches (PP) but retain all lymph nodes. Our analysis of these novel TNF-knockout mice supports the previously disputed notion of the involvement of TNF-TNFR1 signaling in PP organogenesis. Availability of TNF-knockout strains both with and without PP enables more definitive studies concerning the roles of TNF and PP in various immune functions and disease conditions. Here, we report that systemic ablation of TNF, but not the presence of PP per se, is critical for protection against intestinal Listeria infection in mice. PMID- 15832288 TI - Isotope ratio mass spectrometry: delta13C and delta15 N analysis for tracing the origin of illicit drugs. AB - Gas chromatography/combustion/mass spectrometry (GC-C-MS) and elemental analysis/mass spectrometry (EA-MS) techniques are proposed to estimate delta(13)C and delta(15)N values in heroin, morphine, cocaine and hemp leaves, for the purposes of tracing the geographical origins of seized drugs. The values of isotope ratios for pure drugs and drugs with impurities were compared. It was demonstrated that large samples (up to 3 x 10(-6) g C) were combusted completely, so that the results obtained were valid. The data are considered to be an essential supplement to a wide-scale database designed specifically for the delta(13)C and delta(15)N values of drugs. The potential forensic and academic significance of the results is discussed. PMID- 15832289 TI - Humoral immune response to citrullinated collagen type II determinants in early rheumatoid arthritis. AB - Collagen type II (CII) is a relevant joint-specific autoantigen in the pathogenesis of rheumatoid arthritis (RA). Whereas the reasons for the breakage of self tolerance to this major cartilage component are still enigmatic, T cell responses to glycosylated CII determinants in RA patients indicate that post translational modifications play a role. Since the conversion of arginine into citrulline by peptidylarginine deiminases (PAD) in some non-joint-specific antigens such as filaggrin or fibrin has been shown to give rise to RA-specific humoral immune responses, we investigated whether PAD modification of cartilage specific CII might affect its recognition by circulating autoantibodies in early RA. In vitro treatment with purified PAD led to arginine deimination of native CII or of synthetic CII peptides as evidenced by amino acid analysis. The citrullination resulted in modified recognition of the immunodominant CII epitope C1(III) (amino acid residues 359-369) by murine and human antibodies. In a cohort of early RA patients (n=286), IgG antibodies directed toward a synthetic citrullinated C1(III) peptide (citC1(III)-P) were detectable with a prevalence of 40.4%. The partial autoantibody cross-reactivity between citC1(III)-P and citrullinated peptides mimicking epitopes of the cytoskeletal autoantigen filaggrin suggests that autoimmunity to cartilage-specific modified self might be a critical intermediate bridging recognition of PAD-modified extra-articular autoantigens with the disruption of tolerance to native cartilage constituents. PMID- 15832290 TI - CD8+ T cell responses to human immunodeficiency viruses type 2 (HIV-2) and type 1 (HIV-1) gag proteins are distinguishable by magnitude and breadth but not cellular phenotype. AB - The mechanisms underlying the relatively slow progression of human immunodeficiency virus type 2 (HIV-2) compared with HIV-1 infection are undefined and could be a result of more effective immune responses. We used HIV-2 and HIV-1 IFN-gamma enzyme-linked immunospot assays to evaluate CD8(+) T cell responses in antiretroviral-naive HIV-2- ('HIV-2(+)') and HIV-1-infected ('HIV-1(+)') individuals. Gag-specific responses were detected in the majority of HIV-2(+) and HIV-1(+) subjects. Overlapping gag peptide analysis indicated a significantly greater magnitude and breadth of responses in the HIV-1(+) cohort, and this difference was attributable to low responses in HIV-2(+) subjects with undetectable viral load (medians 2107 and 512 spot-forming units per 10(6) PBMC, respectively, p=0.007). We investigated the phenotype of viral epitope-specific CD8(+) T cells identified with HLA-B53- and HLA-B58-peptide tetramers (8 HIV 2(+), 11 HIV-1(+) subjects). HIV-2-specific CD8(+) T cells were predominantly CD27(+) CD45RA(-), and only a minority expressed perforin. The limited breadth and low frequency of CD8(+) T cell responses to HIV-2 gag in aviremic HIV-2(+) subjects suggests that these responses reflect antigen load in plasma, as is the case in HIV-1 infection. Immune control of HIV-2 does not appear to be related to the frequency of perforin-expressing virus-specific CD8(+) T cells. PMID- 15832291 TI - Systematic microanatomical analysis of CXCL13 and CCL21 in situ production and progressive lymphoid organization in rheumatoid synovitis. AB - CXCL13 and CCL21 have been functionally implicated in lymphoid tissue organization both in the upstream phases of lymphoid tissue embryogenesis and in ectopic lymphoid neogenesis in transgenic mice. Here, we analyzed the relationship between CXCL13 and CCL21 production and lymphoid tissue organization in rheumatoid synovitis as a model of a naturally occurring ectopic lymphoneogenesis. Through systematic analysis of mRNA and protein expression, we defined the microanatomical relationship between CXCL13 and CCL21 in progressive aggregational and structural phases of synovial inflammatory infiltrate. We provide the first direct in situ evidence that production of CXCL13 and CCL21 (rather than simply protein binding) is associated with inflammatory lymphoid tissue formation and development with the demonstration, in organized aggregates, of a secondary lymphoid organ-like compartmentalization and vascular association. Notably, the presence of CXCL13 and CCL21 (protein and mRNA) was also demonstrated in non-organized clusters and minor aggregational stages, providing evidence that their induction can take place independently and possibly upstream of T-B compartmentalization, CD21(+) follicular dendritic cell network differentiation and germinal center formation. Our data support the concept that, under inflammatory conditions, CXCL13 and CCL21 participate in lymphoid tissue microanatomical organization, attempting to recapitulate, in an aberrant lymphoid neogenetic process, their homeostatic and morphogenetic physiologic functions. PMID- 15832292 TI - Substance P regulates natural killer cell interferon-gamma production and resistance to Pseudomonas aeruginosa infection. AB - Studies have shown that after Pseudomonas aeruginosa (P. aeruginosa) corneal infection, BALB/c mice that are capable of resolving the disease, locally produce IFN-gamma. As T cells are not detected in the infected cornea of these mice, antibody depletion was used to test whether NK cells produce the cytokine. After depletion, decreased corneal IFN-gamma mRNA and increased disease severity, bacterial load, and PMN infiltrate resulted. Further work determined if substance P (SP), a pro-inflammatory neuropeptide, participated in regulation of this response. To this end, mice were treated with the SP antagonist, spantide I that blocks SP interaction with neurokinin-1, its major receptor. The treatment significantly decreased corneal IFN-gamma and IL-18 protein levels and corneal perforation resulted. In vitro experiments using isolated splenic NK cells confirmed their ability to respond to IL-18 and SP and to secrete IFN-gamma protein. We conclude: that for development of the BALB/c resistance response, NK cells are required to produce IFN-gamma; that the cells express the neurokinin-1 receptor; and that SP directly regulates IFN-gamma production through this receptor. The data suggest a unique link between the nervous system and development of innate immunity in the cornea. PMID- 15832293 TI - Toll-like receptor-dependent activation of several human blood cell types by protamine-condensed mRNA. AB - We reported that RNA condensed on protamine is protected from RNase-mediated degradation and can be used for vaccination. Here, we show that such complexes are also danger signals that activate mouse cells through a MyD88-dependent pathway. Moreover, mRNA-protamine complexes stimulate human blood cells. They strongly activate DC and monocytes, leading to TNF-alpha and IFN-alpha secretion. In addition, protamine-RNA complexes directly activate B cells, NK cells and granulocytes. The detailed analysis of the activated cell types, the study of the cytokines released from PBMC cultured with protamine-RNA complexes and recently published results suggest that TLR-7 and TLR-8 may be involved in the recognition of protamine-stabilized RNA. Our data indicate that protamine-stabilized RNA, which may be similar to RNA condensed in the nucleocapsids of RNA viruses, is a strong danger signal. Thus, similarly to plasmid DNA, protamine-RNA combines antigen production and non-specific immunostimulation. The studies presented here explain the capacity of protamine-RNA to act as a vaccine, and pave the way towards the development of safe and efficient mRNA-based immunotherapies. PMID- 15832294 TI - Implication of allelic polymorphism of osteopontin in the development of lupus nephritis in MRL/lpr mice. AB - Potentially, autoimmune diseases develop from a combination of multiple genes with allelic polymorphisms. An MRL/Mp-Fas(lpr) (/) (lpr) (MRL/lpr) strain of mice develops autoimmune diseases, including lupus nephritis, but another lpr strain, C3H/HeJ-Fas(lpr) (/) (lpr) (C3H/lpr) does not. This indicates that MRL polymorphic genes are involved in the development of the diseases. By quantitative trait loci (QTL) analysis using 527 of the (MRL/lpr x C3H/lpr)F(2) mice, we identified a novel locus for susceptibility to lupus nephritis at map position D5Mit115 on chromosome 5, the same alias of the osteopontin (Opn) gene (LOD score =4.0), susceptible in the MRL allele. In functional analyses of the MRL and C3H Opn alleles using synthetic osteopontin (OPN) made with a new method "cell-free system" with wheat germ ribosomes, the MRL-OPN induced higher expression and production of immunoglobulins as well as cytokines including TNF alpha, IL-1beta and IFN-gamma in splenocytes and/or macrophages than that of the C3H allele. These findings suggest that allelic polymorphism of OPN causes the functional differences in antibody production and macrophage activation between MRL and C3H strains, possibly involved in the development of lupus nephritis. PMID- 15832296 TI - Papillomavirus virus-like particles induce cytokines characteristic of innate immune responses in plasmacytoid dendritic cells. AB - Human papillomavirus (HPV) virus-like particles (VLP) are being extensively tested as vaccines for the prevention of HPV-associated cervical cancer. Dendritic cells (DC) acquire and present antigens, initiating innate and adaptive immune responses. It has been shown previously that DC of the myeloid lineage are capable of generating adaptive immune responses to HPV VLP in vitro. However, the capacity of plasmacytoid DC (pDC) to acquire HPV VLP and the nature of the immune response generated have not been reported. In this study we found that freshly isolated as well as CpG-matured pDC bind papillomavirus VLP and that internalization occurs preferentially in the immature pDC. In contrast to myeloid DC, pDC did not undergo phenotypic maturation upon exposure to HPV16 VLP. However, HPV16 VLP induced pDC to secrete of IFN-alpha and IL-6, both cytokines that play a role in the generation of antibody responses, as well as TNFalpha and IL-8. Given that VLP do not contain viral nucleic acids, these results indicate that viral capsids alone may be capable of inducing cytokine production by pDC. Finally, CpG-activated pDC, but not pDC exposed to HPV16 VLP, activated lymphocytes to secrete IL-10 and low levels of IFN-gamma. Together these findings suggest a possible immunogenic effect of pDC in the setting of VLP vaccination. PMID- 15832297 TI - Development of a new hybrid technique for rapid speciation analysis by directly interfacing a microfluidic chip-based capillary electrophoresis system to atomic fluorescence spectrometry. AB - This paper represents the first study on direct interfacing of microfluidic chip based capillary electrophoresis (chip-CE) to a sensitive and selective detector, atomic fluorescence spectrometry (AFS) for rapid speciation analysis. A volatile species generation technique was employed to convert the analytes from the chip CE effluent into their respective volatile species. To facilitate the chip-CE effluent delivery and to provide the necessary medium for subsequent volatile species generation, diluted HCl solution was introduced on the chip as the makeup solution. The chip-CE-AFS interface was constructed on the basis of a concentric "tube-in-tube" design for introducing a KBH4 solution around the chip effluent as sheath flow and reductant for volatile species generation as well. The generated volatile species resulting from the reaction of the chip-CE effluent and the sheath flow were separated from the reaction mixture in a gas-liquid separator and swept into the AFS atomizer by an argon flow for AFS determination. Inorganic mercury (Hg(II)) and methylmercury (MeHg(I)) were chosen as the targets to demonstrate the performance of the present technique. Both mercury species were separated as their cysteine complexes within 64 s. The precision (relative standard deviation, RSD, n = 5) of migration time, peak area, and peak height for 2 mg.L(-1) Hg(II) and 4 mg.L(-1) MeHg(I) (as Hg) ranged from 0.7 to 0.9%, 2.1 to 2.9%, and 1.5 to 1.8%, respectively. The detection limit was 53 and 161 microg.L( 1) (as Hg) for Hg(II) and MeHg(I), respectively. The recoveries of the spikes of mercury species in four locally collected water samples ranged from 92 to 108%. PMID- 15832295 TI - Impaired IL-4 production by CD8+ T cells in NOD mice is related to a defect of c Maf binding to the IL-4 promoter. AB - CD8(+) T cells play an important role in the induction of the autoimmune response in non-obese diabetic (NOD) mice. Here we describe abnormalities in the control of cytokine production by NOD CD8(+) T cells. NOD CD8(+) T cells had an increased propensity to produce IFN-gamma upon TCR activation, in both adult and 2-week-old mice. NOD CD8(+) T cells had a reduced capacity to produce IL-4 in type 2 conditions compared to CD8(+) T cells from the diabetes-resistant strains BALB/c and C57BL/6. Both GATA-3 and c-Maf, two positive transactivators for IL-4 gene expression, were expressed in type 2 conditions at comparable levels in NOD CD8(+) T cells. The GATA-3 was functional since normal levels of IL-5 were produced and the IL-4 promoter was hyperacetylated in NOD CD8(+) T cells. In contrast, c-Maf failed to bind to its responsive element as determined by chromatin immunoprecipitation (ChIP) assay. These results suggest that NOD CD8(+) T cells possess an increased propensity to produce IFN-gamma and impaired c-Maf dependent DNA binding activities in vivo that lead to reduced IL-4 production following TCR activation. These defects may facilitate the development of the autoimmune response by inducing an overall type 1-biased immune response in NOD mice. PMID- 15832298 TI - Critical role for complement receptor 3 (CD11b/CD18), but not for Fc receptors, in killing of Streptococcus pyogenes by neutrophils in human immune serum. AB - During phagocytosis, surface receptors on neutrophils interact with pathogens opsonized with complement factor C3b/iC3b and in some cases with antibodies. In human immune sera antibodies directed against surface-bound M proteins mediated killing of Streptococcus pyogenes by neutrophils. Surprisingly, blocking of the Fc receptors had little effect on the killing. In contrast, inhibition of C3b/iC3b generation, or blocking of the major neutrophil iC3b receptor CD11b/CD18, enabled S. pyogenes to grow efficiently in immune sera. Inhibition of CD11b/CD18, but not of CD32, the major neutrophil signaling Fc receptor, prevented Streptococcus-induced NADPH oxidase-dependent respiratory burst, and blocking of C3b/iC3b formation inhibited Streptococcus-induced activation of Cdc42, a small GTPase critically involved in transmitting pro-inflammatory signals to the cytoskeleton. Consequently, ligation of CD11b/CD18 by bacteria bound iC3b is necessary for inducing a neutrophil response leading to elimination of S. pyogenes in immune human serum. PMID- 15832299 TI - Determination of dissociation constants of flavonoids by capillary electrophoresis. AB - Ionization constants of some flavanols (catechin and epicatechin) and flavonols (kaempherol, fisetin, morin, and quercetin) are determined by capillary zone electrophoresis (CZE). This technique allows the determination of pK(a) values until about 12. The pK(a) values obtained are compared with those calculated by the SPARC computational program. This program predicts the microscopic and macroscopic pK(a) values and the order of deprotonation of the different -OH groups. While for catechin and epicatechin the first ionizable OH group occurs in ring 1 and the second ionizable group in ring 2, in flavonols the first deprotonation occurs in ring 2 and the second in ring 1. PMID- 15832300 TI - Analysis of oxycodol and noroxycodol stereoisomers in biological samples by capillary electrophoresis. AB - A capillary electrophoresis (CE) method for the separation of the diastereoisomers of 6-oxycodol (6OCOL) and nor-6-oxycodol (N6OCOL), the 6-keto reduced metabolites of oxycodone (OCOD) and noroxycodone (NOCOD), respectively, is reported and employed to assess the stereoselectivity of these metabolic steps in vivo, in vitro, and in chemical synthesis. CE in an untreated fused-silica capillary with acidic buffers containing 2-hydroxypropyl-beta-cyclodextrin, randomly sulfated beta-cyclodextrin, or single isomer heptakis(2,3-diacetyl-6 sulfato)-beta-cyclodextrin (HDAS-beta-CD) is shown to permit the simultaneous separation of the stereoisomers of 6OCOL and N6OCOL. A 100 mM phosphate buffer of pH 2.0 containing 2.05% w/v HDAS-beta-CD provides a medium for rapid analysis and unambiguous identification of these stereoisomers in solid-phase extracts of (i) urines stemming from patients under pharmacotherapy with OCOD, (ii) incubations of OCOD and NOCOD with human liver cytosol and the human liver S9 fraction, and (iii) after chemical synthesis from OCOD and NOCOD using NaBH(4). In all cases, alpha-N6OCOL is shown to be the predominant stereoisomer of N6OCOL. For 6OCOL, the same is true for in vitro formation and for chemical synthesis. In urine, however, beta-6OCOL is observed to be excreted in a higher amount than alpha 6OCOL. For the urinary alpha-/beta-isomer ratio of 6OCOL and N6OCOL, there are no differences between the data obtained for nonhydrolyzed and enzymatically hydrolyzed urines. The data document the stereoselectivity of the 6-keto reduction of OCOD and NOCOD in man. PMID- 15832301 TI - The Debye-Huckel approximation: its use in describing electroosmotic flow in micro- and nanochannels. AB - In this work we consider the electroosmotic flow in a rectangular channel. We consider a mixture of water or other neutral solvent and a salt compound, such as sodium chloride, and other buffers for which the ionic species are entirely dissociated. Results are produced for the case where the channel height is much greater than the width of the electric double layer (EDL) (microchannel) and for the case where the channel height is of the order or slightly greater than the width of the EDL (nanochannel). At small cation, anion concentration differences the Debye-Huckel approximation is appropriate; at larger concentration differences, the Gouy-Chapman picture of the electric double emerges naturally. In the symmetric case, the velocity field and the potential are identical. We specifically focus in this paper on the limits of the Debye-Huckel approximation for a simplified version of a phosphate-buffered saline (PBS) mixture. The fluid is assumed to behave as a continuum and the volume flow rate is observed to vary linearly with channel height for electrically driven flow in contrast to pressure driven flow which varies as height cubed. This means that very large pressure drops are required to drive flows in small channels. However, useful volume flow rates may be obtained at a very low driving voltage. In the course of the solution, we establish the relationship between the wall mole fractions of the electrolytes and the zeta potential. Multivalent electrolyte mixtures are also considered. PMID- 15832302 TI - Enantiomeric separation by capillary electrochromatography using monolithic capillaries with sol-gel-glued cyclodextrin-modified silica particles. AB - By an on-column sol-gel process, a chiral monolithic stationary phase was prepared by the fusion of permethyl-beta-cyclodextrin-silica (Chira-Dex-silica) particles and by linking them to the internal capillary wall. The resulting monolith is stable toward voltage (30 kV) and pressure (300 bar) and possesses a high efficiency (up to 100,000 theoretical plates per meter). Efficient enantiomeric separation of various chiral compounds by pressure-supported capillary electrochromatography (CEC) was achieved. When comparing this method to capillary liquid chromatography (LC) employing the same column in an unified equipment, CEC shows a twofold higher column efficiency at comparable elution times and hence better resolution factors. PMID- 15832303 TI - Comparison of PCR-based mutation detection methods and application for identification of mouse Sult1a1 mutant embryonic stem cell clones using pooled templates. AB - Reverse genetic approaches to generate mutants of model species are useful tools to assess functions of unknown genes. Recent work has demonstrated the feasibility of such strategies in several organisms, exploiting the power of chemical mutagenesis to disrupt genes randomly throughout the genome. To increase the throughput of gene-driven mutant identification, efficient mutation screening protocols are needed. Given the availability of sequence information for large numbers of unknown genes in many species, mutation detection protocols are preferably based on PCR. Using a set of defined mutations in the Hprt1 gene of mouse embryonic stem (ES) cells, we have systematically compared several PCR based point mutation and deletion detection methods available for their ability to identify lesions in pooled samples, which is a major criterion for an efficient large-scale mutation screening assay. Results indicate that point mutations are most effectively identified by heteroduplex cleavage using CEL I endonuclease. Small deletions can most effectively be detected employing the recently described "poison" primer PCR technique. Further, we employed the CEL I assay followed by conventional agarose gel electrophoresis analysis for screening a library of chemically mutagenized ES cell clones. This resulted in the isolation of several clones harboring mutations in the mouse Sult1a1 locus, demonstrating the high-throughput compatibility of this approach using simple and inexpensive laboratory equipment. PMID- 15832304 TI - Study of the electroosmotic flow as a means to propel the mobile phase in capillary electrochromatography in view of further miniaturization of capillary electrochromatography systems. AB - In this paper, we investigate the phenomenon of electroosmosis as a means to propel a mobile phase, in particular in view of an application in microfluidic systems, which are characterized by significantly smaller volumes of the reservoirs and the separation channels compared to conventional instrumentation. In the microfluidic chip, pH changes due to water electrolysis quickly showed an effect on the electroosmotic flow (EOF), which could be counteracted by either regularly exchanging or buffering the mobile phase. Surface treatment was of no effect in regard to EOF stabilization in empty channels but may have an influence in channels filled with a charged monolith. In fused-silica capillaries the EOF was generally found to decrease from 'naked' to surface-treated to monolith filled capillaries. The EOF tended to be higher when an organic solvent (acetonitrile) was added to the mobile phase and could be further increased by substituting the water with equal amounts of methanol. In addition, the hydrostatic pressure exerted by the EOF was investigated. In a microfluidic chip with empty (cross-)channels such an effect could be responsible for a redirection of the flow. In capillaries partially filled with a noncharged (non-EOF generating) monolith, a linear relationship could be established between the EOF created in the empty section of the capillary (apparent mobility) and the length of the monolith (backpressure). In capillaries partially filled with a charged (EOF-producing) monolith, flow inhomogeneities must be expected as a consequence of a superimposition of hydrodynamic pressure and EOF as mobile phase driving force. PMID- 15832305 TI - Genomic rearrangements in the BRCA1 and BRCA2 genes. AB - Mutations in the BRCA1 and BRCA2 genes predispose women to breast and ovarian cancer. BRCA1 and BRCA2 are 83 and 86 kb long, with coding sequences of 5.7 and 10.2 kb, scattered over 22 and 26 coding exons, respectively. The large majority of the alterations identified in these genes are point mutations and small insertions/deletions. However, an increasing number of large genomic rearrangements are being identified, especially in BRCA1. This review gives a brief overview of the techniques used to screen the BRCA1 and BRCA2 genes for large rearrangements, and describes those for which the breakpoints have been characterized. The principal mechanisms that are thought to lead to their formation, founder effects, and recombination hotspots, are also discussed. PMID- 15832306 TI - eOPA1: an online database for OPA1 mutations. AB - Autosomal dominant optic atrophy (ADOA), also known as Kjer disease, is characterized by moderate to severe loss of visual acuity with an insidious onset in early childhood, blue-yellow dyschromatopsia, and central scotoma. An optic atrophy gene, called OPA1, has been identified in most cases of the disease. A total of 83 OPA1 mutations, often family-specific, have been reported so far, and the observations support the hypothesis that haploinsufficiency and the functional loss of a single allele may lead to ADOA. We have developed a new locus-specific database (LSDB), eOPA1 (http://lbbma.univ-angers.fr/eOPA1/) aimed at collecting published and unpublished sequence variations in OPA1. The database has been designed to incorporate new submissions rapidly and will provide a secured online catalog of OPA1 mutations and nonpathogenic sequence variants (NPSVs). The LSDB should prove useful for molecular diagnosis, large-scale mutation statistics, and the determination of original genotype-phenotype correlations in studies on ADOA. PMID- 15832307 TI - A separation-free assay for the detection of mutations: combination of homogeneous time-resolved fluorescence and minisequencing. AB - Single nucleotide primer extension reaction has been widely used in DNA testing, and several detection methods based on this core allelic discrimination have been developed. Most of the reported formats are based on a two step protocol involving first, a liquid phase extension reaction, then a physical separation process (chromatography, electrophoresis, capture on solid support, mass spectrometry). Here we describe a new strategy based on homogeneous time-resolved fluorescence (HTRF), which does not involve any separation process and which allows a simple "mix and measure" protocol. In this approach, a 5'-(europium) cryptate-labeled primer is elongated by a biotinylated dideoxynucleoside triphosphate, followed by the addition of a streptavidin-acceptor conjugate, which gives rise to a long-life fluorescence resonance energy transfer (FRET) signal between the cryptate donor and the acceptor. We present the development of HTRF technology as applied to the diagnosis of tumor suppressor gene p53 (TP53) mutations, and its application to the analysis of genomic DNA from human tumoral samples. The sensitivity of the reported method is compared to the corresponding fluorescent polarization assay. PMID- 15832308 TI - BAC-based PCR fragment microarray: high-resolution detection of chromosomal deletion and duplication breakpoints. AB - The introduction of molecular techniques in conjunction with classical cytogenetic methods has in recent years greatly improved the diagnostic potential for chromosomal abnormalities. In particular, microarray-comparative genomic hybridization (CGH) based on the use of BAC clones promises a sensitive strategy for the detection of DNA copy-number changes on a genomewide scale, offering a resolution as high as >30,000 "bands" (as defined by the number of BACs within the currently highest-density BAC array) [Ishkanian et al., 2004]. We have tested the possibility of further increasing this resolution using PCR fragments generated from individual BAC clones. Using this approach, we have efficiently defined the proximal and distal breakpoints in two cytogenetic cases, one duplication and one deletion, to within 5-20 kb. The results support the potential use of BAC-based PCR fragments to further improve the resolution of the microarray-CGH strategy by an order of magnitude. PMID- 15832311 TI - KinMutBase: a registry of disease-causing mutations in protein kinase domains. AB - A large number of disease-causing mutations have been identified from several protein kinases. KinMutBase is a comprehensive knowledge base for human disease related mutations in protein kinase domains (http://bioinf.uta.fi/KinMutBase/). The latest version contains 582 different mutations for 1,790 cases in 1,322 families. KinMutBase entries are described on the DNA, mRNA, and protein level. Numbers for affected patients and families are also provided. KinMutBase has extensive amount of links and cross-references to literature, other databases, and information sources. There are numerous interactive pages about sequences, structures, mutation statistics, and diseases. Detailed statistical study was done on frequencies of different types of mutations both on the DNA and protein level in serine/threonine kinase (PSK) and tyrosine kinase (PTK). Three dimensional structures indicate clustering of disease-related mutations mainly to conserved subdomains, and substrate and coligand binding amino acids, although mutations appear throughout the sequences. CpG containing codons, especially for arginine, constitute the majority of mutational hotspots. There are certain clear differences in mutation patterns and types between PSKs and PTKs. PMID- 15832309 TI - Ancient origin of the CAG expansion causing Huntington disease in a Spanish population. AB - Huntington disease (HD) is an autosomal dominant neurodegenerative disorder characterized clinically by progressive motor impairment, cognitive decline, and emotional deterioration. The disease is caused by the abnormal expansion of a CAG trinucleotide repeat in the first exon of the huntingtin gene in chromosome 4p16.3. HD is spread worldwide and it is generally accepted that few mutational events account for the origin of the pathogenic CAG expansion in most populations. We have investigated the genetic history of HD mutation in 83 family probands from the Land of Valencia, in Eastern Spain. An analysis of the HD/CCG repeat in informative families suggested that at least two main chromosomes were associated in the Valencian population, one associated with allele 7 (77 mutant chromosomes) and one associated with allele 10 (two mutant chromosomes). Haplotype A-7-A (H1) was observed in 47 out of 48 phase-known mutant chromosomes, obtained by segregation analysis, through the haplotype analysis of rs1313770 HD/CCG-rs82334, as it also was in 120 out of 166 chromosomes constructed by means of the PHASE program. The genetic history and geographical distribution of the main haplotype H1 were both studied by constructing extended haplotypes with flanking short tandem repeats (STRs) D4S106 and D4S3034. We found that we were able to determine the age of the CAG expansion associated with the haplotype H1 as being between 4,700 and 10,000 years ago. Furthermore, we observed a nonhomogenous distribution in the different regions associated with the different extended haplotypes of the ancestral haplotype H1, suggesting that local founder effects have occurred. PMID- 15832310 TI - Quantitative analysis of SMN1 and SMN2 genes based on DHPLC: a highly efficient and reliable carrier-screening test. AB - Autosomal recessive spinal muscular atrophy (SMA) is a common, fatal neuromuscular disease caused by homozygous absence of the SMN1 gene in approximately 94% of patients. However, a highly homologous SMN2 gene exists in the same chromosome interval, centromeric to SMN1, and hampers detection of SMN1. We present a new, rapid, simple, and highly reliable method for detecting the SMN1 deletion/conversion and for determining the copy numbers of the SMN1 and SMN2 genes by DHPLC. We analyzed SMN1/SMN2 gene exon 7 deletion/conversion by DHPLC. A total of 25 patients with spinal muscular atrophy lacking the SMN1 gene as well as 309 control individuals from the general population and the family members of patients with SMA were analyzed. By DHPLC analysis, we could detect the SMA-affected cases efficiently just by recognizing an SMN2-only peak. Furthermore, after specific primer amplification and adjustment of the oven temperature, all of the SMA carriers with an SMN1/SMN2 ratio not equal to 1 could be identified unambiguously by this simple and efficient detection system. To calculate the total SMN1/SMN2 gene dosages further, we developed a specific multiplex competitive PCR protocol by simultaneously amplifying the CYBB gene (X linked), the KRIT1 gene (on chromosome arm 7q), and the SMN1/SMN2 gene ratio by DHPLC. By applying this technique, we could successfully designate all of the genotypes with different SMN1/SMN2 gene copy numbers, including equal and unequal amounts of SMN1 and SMN2. We demonstrated that DHPLC is a fast and reliable tool for detection of carriers of SMA. PMID- 15832312 TI - Population spectrum of ACADM genotypes correlated to biochemical phenotypes in newborn screening for medium-chain acyl-CoA dehydrogenase deficiency. AB - Medium-chain acyl-CoA dehydrogenase deficiency (MCADD) is the most frequent inherited defect of fatty acid oxidation, with a significant morbidity and mortality in undiagnosed patients. Adverse outcomes can effectively be prevented by avoiding metabolic stress and following simple dietary measures. Therefore, prospective newborn screening (NBS) is being proposed for this condition. However, technical validation of MCADD population screening and assessment of its overall benefit require broadening of the as-yet-scarce knowledge of the MCADD genetic heterogeneity unraveled by NBS and its phenotypic consequences. Here, we describe the entire spectrum of sequence variations occurring in newborns with MCADD in the population of Bavaria, Germany, in relation to the biochemical phenotype. Among 524,287 newborns, we identified 62 cases of MCADD, indicating a birth incidence of 1 in 8,456. In all of the 57 newborns available for analysis, two alterations within the MCADD gene (ACADM) were identified. The most prevalent alteration c.985A>G (Lys329Glu) occurred in 27 (47%) newborns in the homozygous and in 18 (32%) in the heterozygous state (63% of defective alleles). The mild folding variant c.199T>C (Tyr67His) was identified in nine individuals, six of them being compound heterozygous with c.985A>G (Lys329Glu). Neither of the prevalent alterations were found in the remaining nine newborns. A total of 18 sequence variations were identified; 13 of them were novel: eight missense mutations, one nonsense mutation, two splice variants, and two small deletions. The remaining five were previously reported in MCADD patients. The ACADM heterogeneity uncovered was larger as anticipated from previous c.985A>G (Lys329Glu) carrier screening data. In addition, we show that MCADD appears to occur as frequently in Turkish newborns as in the native German population. Our data validate that biochemical NBS for MCADD is a highly specific procedure for disease detection, with the identification of a significant share of milder biochemical phenotypes, such as c.199T>C (Tyr67His). These show statistically lower acylcarnitine markers, allowing us to distinguish subgroups within the spectrum of ACADM sequence variations that correlate to biochemical MCADD disease expression. Our data might provide technical and medical guidance for decision making in the worldwide efforts to introduce MCADD population screening. PMID- 15832313 TI - TCOF1 mutation database: novel mutation in the alternatively spliced exon 6A and update in mutation nomenclature. AB - Recently, a novel exon was described in TCOF1 that, although alternatively spliced, is included in the major protein isoform. In addition, most published mutations in this gene do not conform to current mutation nomenclature guidelines. Given these observations, we developed an online database of TCOF1 mutations in which all the reported mutations are renamed according to standard recommendations and in reference to the genomic and novel cDNA reference sequences (www.genoma.ib.usp.br/TCOF1_database). We also report in this work: 1) results of the first screening for large deletions in TCOF1 by Southern blot in patients without mutation detected by direct sequencing; 2) the identification of the first pathogenic mutation in the newly described exon 6A; and 3) statistical analysis of pathogenic mutations and polymorphism distribution throughout the gene. PMID- 15832315 TI - Characterization of iduronate-2-sulfatase gene-pseudogene recombinations in eight patients with Mucopolysaccharidosis type II revealed by a rapid PCR-based method. AB - Various types of complex genetic rearrangements involving the iduronate-2 sulfatase (IDS) and its homologous pseudogene (IDS2, IDSP1) have so far been reported as the cause of Mucopolysaccharidosis type II (MPS2 or MPS II; Hunter syndrome). When using conventional mutational analyses, the occurrence in intronic regions of these rearrangements can be misleading. Here, we describe a rapid PCR-based method set up to detect possible gene/pseudogene recombinations among a series of Italian male patients who had negative results in the mutation analysis of the IDS gene. Our approach selected eight unrelated patients showing recombinations. The characterization of the proximal regions containing the breakpoints in the eight patients identified four different rearrangements due to both inversion and conversion events. Comparison of our data with previous publications confirmed that the recombinations between the IDS gene and the IDS2 pseudogene result from separate events, considering their occurrence at different positions within the same "hotspot" genomic region in unrelated patients. The RT PCR analysis of the available cDNAs pointed out the different effects of similar rearrangements on the expression of the IDS gene. This method can be utilized effectively in the absence of the patients' cDNA, as well as for carrier detection among female family members. This advantageous approach reduces costs, is less time-consuming, and requires a smaller DNA quantity in comparison to the Southern blot hybridization technique often utilized for such complex rearrangements. PMID- 15832314 TI - Phospholipid transfer protein (PLTP) mRNA expression is stimulated by developing embryos in the oviduct. AB - In mammal, fertilization and early preimplantation embryo development occurs in the oviduct. Evidence is accumulating that the oviductal epithelia secrete various biomolecules to the lumen during the secretory phase of the estrus cycle to enhance embryo development. This secretory activity of the oviduct is under the regulation of steroid hormones. Observations also suggested that the gametes and embryos modulate the physiology and gene-expressing pattern of the oviduct. However, the underlying molecular changes remain elusive. We hypothesize that the developing embryos interact with the surrounding environment and affect the gene expression patterns of the oviduct, thereby modulating the oviductal secretory activity conducive to the preimplantation embryo development. To test this hypothesis, suppression subtractive hybridization (SSH) was used to compare the gene expressions in mouse oviduct containing transferred in vitro cultured preimplantation embryos with that of oviduct containing oocytes during the preimplantation period. We reported here the identification and characterization of phospholipids transfer protein (PLTP), which is highly expressed in the embryo containing oviduct and localized at the oviductal epithelium by in situ hybridization. PLTP contains signal peptide putative for secretory function. More importantly, PLTP mRNA increases in the oviductal epithelia of pregnant, but not pseudo-pregnant mice when assayed by real-time PCR. Taken together, our data suggested that PLTP may play important role(s) during in vivo preimplantation embryo development. This molecule would be a target to delineate the mechanisms and the roles of oviductal secretory proteins on early embryonic development. PMID- 15832316 TI - Source density-driven independent component analysis approach for fMRI data. AB - Independent component analysis (ICA) has become a popular tool for functional magnetic resonance imaging (fMRI) data analysis. Conventional ICA algorithms including Infomax and FAST-ICA algorithms employ the underlying assumption that data can be decomposed into statistically independent sources and implicitly model the probability density functions of the underlying sources as highly kurtotic or symmetric. When source data violate these assumptions (e.g., are asymmetric), however, conventional ICA methods might not work well. As a result, modeling of the underlying sources becomes an important issue for ICA applications. We propose a source density-driven ICA (SD-ICA) method. The SD-ICA algorithm involves a two-step procedure. It uses a conventional ICA algorithm to obtain initial independent source estimates for the first-step and then, using a kernel estimator technique, the source density is calculated. A refitted nonlinear function is used for each source at the second step. We show that the proposed SD-ICA algorithm provides flexible source adaptivity and improves ICA performance. On SD-ICA application to fMRI signals, the physiologic meaningful components (e.g., activated regions) of fMRI signals are governed typically by a small percentage of the whole-brain map on a task-related activation. Extra prior information (using a skewed-weighted distribution transformation) is thus additionally applied to the algorithm for the regions of interest of data (e.g., visual activated regions) to emphasize the importance of the tail part of the distribution. Our experimental results show that the source density-driven ICA method can improve performance further by incorporating some a priori information into ICA analysis of fMRI signals. PMID- 15832317 TI - Adsorption of 6-mercaptopurine and 6-mercaptopurine-ribosideon silver colloid: a pH-dependent surface-enhanced Raman spectroscopy and density functional theory study. II. 6-mercaptopurine-riboside. AB - Surface-enhanced Raman spectroscopy (SERS) has been applied to characterize the interaction of 6-mercaptopurine-ribose (6MPR), an active drug used in chemotherapy of acute lymphoblastic leukemia, with a model biological substrate at therapeutic concentrations and as function of the pH value. Therefore, a detailed vibrational analysis of crystalline and solvated (6MPR) based on Density Functional Theory (DFT) calculations of the thion and thiol tautomers has been performed. 6MPR adopts the thion tautomeric form in the polycrystalline state. The SERS spectra of 6MPR and 6-mercaptopurine (6MP) recorded on silver colloid provided evidence that the ribose derivative shows different adsorption behavior compared with the free base. Under acidic conditions, the adsorption of 6MPR on the metal surface via the N7 and possibly S atoms was proposed to have a perpendicular orientation, while 6MP is probably adsorbed through the N9 and N3 atoms. Under basic conditions both molecules are adsorbed through the N1 and possibly S atoms, but 6MP has a more tilted orientation on the silver colloidal surface while 6MPR adopts a perpendicular orientation. The reorientation of the 6MPR molecule on the surface starts at pH 8 while in the case of 6MP the reorientation starts around pH 6. Under basic conditions, the presence of the anionic molecular species for both molecules is suggested. The deprotonation of 6MP is completed at pH 8 while the deprotonation of the riboside is finished at pH 10. For low drug concentrations under neutral conditions and for pH values 8 and 9, 6MPR interacts with the substrate through both N7 and N1 atoms, possibly forming two differently adsorbed species, while for 6MP only one species adsorbed via N1 was evidenced. PMID- 15832318 TI - The development of spatial capacity in piloting and dead reckoning by infant rats: use of the huddle as a home base for spatial navigation. AB - Two forms of spatial navigation, piloting using external cues and dead reckoning using self-movement cues, are manifest in the outward and homeward trips of adult rats exploring from a home base. Here, the development of these two forms of spatial behavior are described for rats aged 14-65 days using a new paradigm in which a huddle of pups or an artificial huddle, a small heat pad, served as a home base on an open circular table that the rats could explore. When moving away from both home bases, the travel distance, path complexity, and number of stops of outward trips from the home base increased progressively with age from postnatal day 16 through 22. When returning to the home bases, the return trips to the home base were always more direct and had high travel velocities even though travel distance increased with age for the longest trips. The results are discussed in relation to the ideas that: (1) the pups pilot on the outward portion of their excursion and dead reckon on the homeward portion of their excursion, and (2) the two forms of navigation and associated spatial capacity are interdependent and develop in parallel and in close association with locomotor skill. PMID- 15832319 TI - Handedness and specific language impairment: a study of 6-year-old twins. AB - Handedness and language skills were assessed in 196 same-sex twin pairs (101 MZ and 95 DZ), who were selected from an epidemiological study of twins, so that children with risk of language impairment were over-represented. When assessed at 6 years of age, 83 children met criteria for specific language impairment (SLI), 32 had general developmental (GD) delay, and the remaining 277 were typically developing (TD). Hand preference (HP) assessed by inventory did not distinguish SLI, TD, or GD groups. The quantification of hand preference (QHP) measure, which measures persistence of a HP when reaching across the midline, did show weaker HP in those with SLI compared to the other two groups. It is suggested that the QHP measure assesses developmental aspects of manual lateralization, and is sensitive to neurodevelopmental immaturity in SLI. Furthermore, genetic analysis showed that the QHP measure, unlike the handedness inventory, was significantly heritable. PMID- 15832320 TI - Social withdrawal behaviors in nonhuman primates and changes in neuroendocrine and monoamine concentrations during a separation paradigm. AB - This study investigated relationships between withdrawal behaviors in rhesus macaques and changes in monoamine metabolite and endocrine concentrations during repeated psychosocial stress. Rhesus monkeys (N = 71) experienced maternal separation in which four separations took place during four consecutive weeks. Behavioral observations were made, as well as plasma concentrations of cortisol and cerebrospinal fluid concentrations of the serotonin, dopamine, and norepinephrine metabolites were obtained. Animals were assigned to high, moderate, and low withdrawal groups, defined using baseline durations of withdrawal behaviors. Highly withdrawn animals showed less reduction than nonwithdrawn animals in serotonin metabolite concentrations over repeated separations. Highly withdrawn macaques also failed to significantly reduce cortisol concentrations across separation weeks. More adaptation in central serotonin functioning and cortisol concentrations was seen in nonwithdrawn primates than in highly withdrawn primates; these findings have implications for increased risk of developing anxiety disorders in highly inhibited children. PMID- 15832321 TI - Early learning failure impairs adult learning in rats. AB - Early life experiences may affect adult learning ability. In two experiments we tested the effect of early learning failure on adult performance in Wistar rats. In the first experiment 17-day-old rats (PN17), but not 25-day-old rats (PN25), trained in a hidden platform water maze task showed deficits in tone-shock avoidance learning when they were 3-months-old. The second experiment, which included random-platform and non-platform control groups, confirmed the effect of early (PN18) spatial learning failure on adult avoidance learning. However, post weaning training (PN25) without platform also tended to induce adult learning deficits as long as the adult task difficulty was increased. The older non platform group did not differ from the impaired group which received early training in a fixed hidden platform task. The results are discussed in terms of the relevance of early learning outcome and developmental stage on adult general learning deficits which may be related to the learned helplessness phenomenon and developmental neural plasticity. PMID- 15832322 TI - Vagal regulation and emotional intensity predict children's sleep problems. AB - We examined the role of children's emotional intensity and vagal functioning in predicting sleep problems in healthy elementary school-aged children. Children's dispositional emotionality was examined via parent report, and their vagal regulation was assessed via respiratory sinus arrhythmia (RSA) during a baseline and a reaction time (RT) task. Sleep problems were examined through both child reports, and home monitoring with wrist actigraphs for four consecutive nights. Increased emotional intensity was predictive of a reduced amount of sleep and increased night activity. Less apt vagal regulation, characterized by lower levels of RSA suppression to the RT task, was predictive of increased sleep problems as assessed through both subjective and actigraphy-based measures of sleep. Results indicate that children's emotionality and regulation predict unique variance in the amount and quality of children's sleep, and suggest that they may underlie, at least in part, sleep disturbances in healthy children. PMID- 15832323 TI - Rearing environment and hypothalamic-pituitary-adrenal regulation in young rhesus monkeys (Macaca mulatta). AB - A mammal's early social environment has important regulatory effects on its behavior and physiology, and this is especially true for regulation of the hypothalamic-pituitary-adrenal (HPA) system. The present study was designed to test hypotheses that various aspects of the social environment are important influences on HPA regulation. Seven hundred seventy eight, 3- to 4-month-old rhesus monkeys were studied as part of a standardized, 24-hr biobehavioral assessment program, which included blood sampling to determine plasma cortisol concentrations. Results indicate that nursery-rearing results in a reduced cortisol set-point for the HPA system, and, for nursery-reared (NR) animals, more peer exposure during infancy is associated with a higher set-point. Age and sex differences during this period were evident but small in magnitude. These data demonstrate the important regulatory role of the social environment on nonhuman primate physiology and suggest caution in assuming that differences between individuals' cortisol levels reflect only differences in perceptions of the "stressfulness" of events. PMID- 15832324 TI - The effects of poly(ethylene glycol) on the solution structure of human serum albumin. AB - Protein physical and chemical properties can be altered by polymer interaction. The presence of several high affinity binding sites on human serum albumin (HSA) makes it a possible target for many organic and polymer molecules. This study was designed to examine the interaction of HSA with poly(ethylene glycol) (PEG) in aqueous solution at physiological conditions. Fourier transform infrared, ultraviolet-visible, and CD spectroscopic methods were used to determine the polymer binding mode, the binding constant, and the effects of polymer complexation on protein secondary structure. The spectroscopic results showed that PEG is located along the polypeptide chains through H-bonding interactions with an overall affinity constant of K = 4.12 x 10(5) M(-1). The protein secondary structure showed no alterations at low PEG concentration (0.1 mM), whereas at high polymer content (1 mM), a reduction of alpha-helix from 59 (free HSA) to 53% and an increase of beta-turn from 11 (free HSA) to 22% occurred in the PEG-HSA complexes (infrared data). The CDSSTR program (CD data) also showed no major alterations of the protein secondary structure at low PEG concentrations (0.1 and 0.5 mM), while at high polymer content (1 mM), a major reduction of alpha-helix from 69 (free HSA) to 58% and an increase of beta-turn from 7 (free HSA) to 18% was observed. PMID- 15832325 TI - Individual differences in biobehavioral organization during problem-solving in toddlers: the influence of maternal behavior, infant-mother attachment, and behavioral inhibition on the attachment-exploration balance. AB - The aim of the study was to investigate the influence of maternal behavior, infant-mother attachment, and behavioral inhibition on the biobehavioral organization of toddlers during a primary non-attachment-related challenging context. In a sample of 76 toddlers, quality of attachment (secure, avoidant, ambivalent, disorganized) was assessed at 12 months. At 22 months, the subjects were observed in a challenging task according to [Matas et al. (1978) Child Development 49: 547-556], and salivary cortisol was assessed before and afterwards. Behavioral analyses included child task orientation and exploration, help seeking, proximity seeking, and attachment-exploration-balance as well as maternal supportive presence and quality of assistance. Behavioral inhibition was assessed from maternal reports. Findings from concurrent analyses indicate relations between toddler behavior and adrenocortical function and the importance of concurrent maternal support for toddler behavior. Longitudinal analyses showed that quality of attachment predicted task orientation, help seeking behavior, and the quality of the attachment-exploration-balance, while behavioral inhibition was related to approach to the task. In line with previous findings, adrenocortical activation was found only in insecure infants with high behavioral inhibition indicating the function of attachment security as a social buffer against less adaptive temperamental dispositions. Finally, within-group analyses indicated a social regulation pattern in secure toddlers while the behavior of insecure ones was additionally linked to behavioral inhibition and physiological reactivity. PMID- 15832326 TI - Live 3D echo guidance of catheter-based endomyocardial injection. AB - Local delivery of therapeutic agents into the myocardium is limited by suboptimal imaging. We evaluated the feasibility and accuracy of live 3D echo to guide left ventricular endomyocardial injection. An intramyocardial injection catheter was positioned in the left ventricle in five healthy Yorkshire pigs using fluoroscopy. All other catheter manipulations were performed with live biplane and 3D echo guidance. In each animal, a total of 12 endomyocardial injections (volume, 50-100 microl) of echo contrast mixed with blue tissue dye were performed. Four injections, 10 mm apart, were directed to three myocardial target zones: the anterior septum at the mitral valve level (zone 1); the posterolateral wall between the heads of the papillary muscles (zone 2); and the apex (zone 3). The injections were aimed to form a transverse line in zones 1 and 2 and an inverted triangular pyramid in zone 3. The animals were sacrificed, the hearts were inspected and the left ventricular endocardium was examined to create a map of injection marks. Success, defined as a visible injection of tissue dye, was 95%, and accuracy, defined as an injection into the target zone, was 83%. There was no significant difference in accuracy between the zones. Live 3D echo can successfully guide endomyocardial injections by accurately targeting specific myocardial zones, verifying catheter apposition and, when combined with echo contrast, providing real-time visualization of injectate deposition. PMID- 15832327 TI - Gross anatomy of the interphalangeal joint of the great toe: implications for excision of plantar capsular accessory ossicles. AB - Bony or cartilaginous ossicles occur at the plantar aspect of the interphalangeal joint of the great toe. The variation in pattern, prevalence, and anatomic relationships of these structures is not clearly established in the literature, especially in a Caucasian population. Without this knowledge, pathology at this joint may be underestimated and surgical approaches may be poorly planned particularly as radiographs underestimate the incidence of ossicles at this joint. The aims of this study were to determine the incidence and pattern of ossicles at this joint and to establish their anatomic relationships to aid planning the approach for their excision. The interphalangeal joint of the left hallux was dissected in 40 British Caucasian cadavers and the pattern of nodules and their anatomic relationships were established. In 27.5% of subjects, there was no identifiable ossicle and, in these specimens, the tendon of flexor hallucis longus was adherent to the joint capsule. In the remaining specimens (72.5%), a bursa separated the tendon of flexor hallucis longus from the plantar joint capsule and nodules were found embedded within the joint capsule. More than half (52.5%) of the specimens had a single nodule located centrally within the plantar capsule and the remaining 20% had two nodules lying within the capsule. This study shows that a large proportion of the population have either one or two bony or cartilaginous ossicles at this joint. It has also shown that, when present, these structures do not lie within the tendon of flexor hallucis longus and may be most safely excised from a medial approach. PMID- 15832328 TI - Coexistence of multiple anomalies in the carpal tunnel. AB - We determined the frequency of anomalous structures within the carpal tunnels of 89 cadaveric forearm-hand specimens. We also examined these same specimens for variations in the branching pattern of the median nerve, and analyzed the range in length and width of the lumbricals. Many of the hands contained extra tendinous slips from the long flexors within the tunnel, subligamentous thenar branches of the median nerve, or lumbricals with bipennate origins. Only one hand had an anomalous muscle belly within the tunnel, two had persistent median arteries, two had high division of the median nerve in the distal forearm, and eight had lumbricals with lengths or widths that were greater or less than 2 standard deviations (SD) from the mean. Twenty-nine percent of all hands examined had two to five anomalies/variations per tunnel, whereas another 27% had one anomaly or variation per tunnel. More right hands (17%) than left (11%) contained two to five anomalous/variant structures per carpal tunnel. More right hands (19%) than left (8%) contained only one variant/anomalous structure per carpal tunnel. Anticipation of the frequency and multiplicity of anomalous structures and variations within this region is of importance to clinicians, particularly surgeons. PMID- 15832329 TI - No effect of 85 mT permanent magnets on laser-Doppler measured blood flow response to inspiratory gasps. AB - Although no effects of permanent magnets on resting skin blood flow (SBF) in humans have yet been demonstrated, the possibility that magnet related effects might modify dynamic SBF changes has not been previously studied. We hypothesized that magnets may alter local neurovascular mechanisms to cause changes in normal SBF vasoactive responses. To test this, we studied the effects of a magnet on SBF reductions induced by sympathetic reflexes associated with deep inspirations. SBF was continuously monitored by a dual channel laser-Doppler flowmeter with probes on the middle finger dorsum of both hands of 24 healthy subjects. In the first of two successive intervals, each of the fingers rested on sham ceramic magnets (control interval). Subsequently, one finger rested on an active magnet and the other finger on a sham (experimental interval). Skin temperatures were also measured. The magnet was a 37 mm diameter x 14 mm thick ceramic magnet with a surface field strength of 85 mT measured in the geometrical center of the magnet. Field strength at the finger dorsum, 13 mm above magnet, was 31.5 mT. During each interval, three deep breaths were used to elicit SBF reductions. Responses were calculated as the percent reduction in SBF from its prior 20 s average. Breaths in each interval were spaced 3 min apart to permit full recovery between responses. The experimental interval started after an active magnet was in place for 20 min. Results showed no significant difference in either vasoconstrictive responses or skin temperature due to the magnet. We conclude that magnets of the type, strength and duration used, have no significant effect on vasoconstrictive processes associated with this sympathetic reflex in this group of healthy subjects. PMID- 15832330 TI - Devices for gradient static magnetic field exposure. AB - We describe devices designed for magnetic field exposures in which field amplitude and gradients are controlled simultaneously. Dosimetry based on field continuation of high resolution magnetic field scans and numerical models is compared with validation measurements. The dosimetry variables we consider are based on the assumption that the biological or chemical system under study has field transducers that are spatially isotropic, so that absolute field amplitude and two gradient components fully describe local exposure. PMID- 15832331 TI - Low frequency therapeutic EMF differently influences experimental muscle pain in female and male subjects. AB - Effects of a pulsating, half sine wave magnetic field (MF) with a frequency of 100 pps and 15 mT rms flux density, generated by the MD TEMF device (EMF Therapeutics, Inc., Chattanooga), on subjective pain rating, heart rate, and arterial blood pressure were tested in a double blind, crossover design study employing experimental muscle pain. Each of 24 healthy volunteers (12 females and 12 males, 24.7 +/- 3.2 years of age) received painful stimulation induced by the infusion of 5% hypertonic saline (HS) into the erector spinae muscle during real and sham MF exposure, in counterbalanced order. Exposure to MF differently affects subjective pain estimates in females and males. MF exposure increased averaged pain level and time integral of pain ratings in females, whereas no statistically significant difference for these characteristics was found in males. Pain related elevation of systolic and diastolic blood pressure was observed during both real and sham EMF exposure in female and male subjects. PMID- 15832332 TI - Molecular change signal-to-noise criteria for interpreting experiments involving exposure of biological systems to weakly interacting electromagnetic fields. AB - We describe an approach to aiding the design and interpretation of experiments involving biological effects of weakly interacting electromagnetic fields that range from steady (dc) to microwave frequencies. We propose that if known biophysical mechanisms cannot account for an inferred, underlying molecular change signal-to-noise ratio, (S/N)gen, of a observed result, then there are two interpretation choices: (1) there is an unknown biophysical mechanism with stronger coupling between the field exposure and the ongoing biochemical process, or (2) the experiment is responding to something other than the field exposure. Our approach is based on classical detection theory, the recognition that weakly interacting fields cannot break chemical bonds, and the consequence that such fields can only alter rates of ongoing, metabolically driven biochemical reactions, and transport processes. The approach includes both fundamental chemical noise (molecular shot noise) and other sources of competing chemical change, to be compared quantitatively to the field induced change for the basic case that the field alters a single step in a biochemical network. Consistent with pharmacology and toxicology, we estimate the molecular dose (mass associated with field induced molecular change per mass tissue) resulting from illustrative low frequency field exposures for the biophysical mechanism of voltage gated channels. For perspective, we then consider electric field-mediated delivery of small molecules across human skin and into individual cells. Specifically, we consider the examples of iontophoretic and electroporative delivery of fentanyl through skin and electroporative delivery of bleomycin into individual cells. The total delivered amount corresponds to a molecular change signal and the delivery variability corresponds to generalized chemical noise. Viewed broadly, biological effects due to nonionizing fields may include animal navigation, medical applications, and environmental hazards. Understanding necessary conditions for such effects can be based on a unified approach: quantitative comparison of the estimated chemical change due to a particular electromagnetic field exposure to that due to competing influences, with both estimates based on a biophysical mechanism model within the context of a model of a biological system. PMID- 15832333 TI - Time dependent modifications of Hep G2 cells during exposure to static magnetic fields. AB - Morphological modifications, i.e., cell shape, cell surface sugar residues, cytoskeleton, and apoptosis of Hep G2 cells during 24 h exposure to 6 mT static magnetic field (static MF) were studied by means of light and electron microscopy and cytochemistry. Progressive modifications of cell shape and surface were observed during the entire period of exposure to static MF. Control cells were polyhedric with short microvilli covering the cell surface, while those exposed to static MF, were elongated with many irregular microvilli randomly distributed on the cell surface. At the end of the exposure period, the cells had a less flat shape due to partial detachment from the culture dishes. However, throughout the period of exposure under investigation, the morphology of the organelles remained unmodified and cell proliferation was only partially affected. In parallel with cell shape changes, the microfilaments and microtubules, as well as the quantity and distribution of surface ConA-FITC and Ricinus communnis-FITC labeling sites, were modified in a time dependent manner. Apoptosis, which was almost negligible at the beginning of experiment, increased to about 20% after 24 h of continuous exposure. The induction of apoptosis was likely due to the increment of [Ca2+]i during exposure. In conclusion, the data reported in the present work indicates that 6 mT static MF exposure exerts time dependent biological effects on Hep G2 cells. PMID- 15832334 TI - Comparison perception of singular transcranial magnetic stimuli by subjectively electrosensitive subjects and general population controls. AB - Transcranial magnetic stimulation of the dorsolateral prefrontal cortex by single pulses of varying field intensities was used to measure thresholds of individual perception and motor response in three groups of subjects: subjectively electrosensitive people, general population controls with a high burden of complaints related to electromagnetic field (EMF) exposure in the literature (highest decile in complaint burden), and general population controls with a low burden of complaints (lowest decile in complaint burden). The major study endpoint was the ability of the subjects to differentiate between real magnetic stimulation and a sham condition. There were no significant differences between groups in the thresholds, neither of detecting the real magnetic stimulus nor in motor response. But the three groups differed significantly in differentiating between stimulation and sham condition, with the subjectively electrosensitive people having the lowest ability to differentiate and the control group with high level of EMF-related complaints having the best ability to differentiate. Differences between groups were mostly due to false alarm reactions in the sham condition reported by subjectively electrosensitives (SES). We found no objective correlate of the self perception of being "electrosensitive." Overall, our experiment does not support the hypothesis that subjectively electrosensitive patients suffer from a physiological hypersensitivity to EMFs or stimuli. Further research should focus on disposing factors explaining the unspecific sensory hyperresponsiveness of subjectively electrosensitive subjects. PMID- 15832335 TI - No influence on selected parameters of human visual perception of 1970 MHz UMTS like exposure. AB - In recent years several studies regarding possible effects of radio frequency (RF) electromagnetic fields (EMFs) on cognitive brain function were reported. In many of these studies on awake humans the working tasks were presented visually to the test subjects, e.g., on a computer screen. Therefore, the question of where in the chain of visual perception, brain processing and response a possible effect could be induced seems to be of interest. In this study, possible effects of exposure to a generic 1.97 GHz UMTS-like signal on human visual perception were investigated in a double blinded, crossover study including 58 healthy volunteer subjects (29 male, 29 female), aged 29 +/- 5.1 years (mean +/- SD). Each test subject underwent a battery of four different clinical tests three times (two different exposure levels and sham exposure) to assess selected parameters of visual perception. The generic signals applied to the subjects' head represented the RF emissions of an UMTS mobile phone under constant receiving conditions and the under condition of strongly varying transmit power, i.e., the signal envelope contained low frequency components. In the high exposure condition the resulting average exposure of the test subjects in the cortex of the left temporal lobe of the brain was 0.63 W/kg (1 g averaged SAR) and 0.37 W/kg (10 g averaged SAR). Low exposure condition was one tenth of high exposure and sham was at least 50 dB (corresponding to a factor of 100,000) below low exposure. Statistical evaluation of the obtained test results revealed no statistically significant differences in the investigated parameters of visual perception between the exposure conditions and sham exposure. PMID- 15832336 TI - Evaluation of genotoxic effects in human peripheral blood leukocytes following an acute in vitro exposure to 900 MHz radiofrequency fields. AB - Human peripheral blood leukocytes from healthy volunteers have been employed to investigate the induction of genotoxic effects following 2 h exposure to 900 MHz radiofrequency radiation. The GSM signal has been studied at specific absorption rates (SAR) of 0.3 and 1 W/kg. The exposures were carried out in a waveguide system under strictly controlled conditions of both dosimetry and temperature. The same temperature conditions (37.0 +/- 0.1 degrees C) were realized in a second waveguide, employed to perform sham exposures. The induction of DNA damage was evaluated in leukocytes by applying the alkaline single cell gel electrophoresis (SCGE)/comet assay, while structural chromosome aberrations and sister chromatid exchanges were evaluated in lymphocytes stimulated with phytohemagglutinin. Alterations in kinetics of cell proliferation were determined by calculating the mitotic index. Positive controls were also provided by using methyl methanesulfonate (MMS) for comet assay and mitomycin-C (MMC), for chromosome aberration, or sister chromatid exchange tests. No statistically significant differences were detected in exposed samples in comparison with sham exposed ones for all the parameters investigated. On the contrary, the positive controls gave a statistically significant increase in DNA damage in all cases, as expected. Thus the results obtained in our experimental conditions do not support the hypothesis that 900 MHz radiofrequency field exposure induces DNA damage in human peripheral blood leukocytes in this range of SAR. PMID- 15832337 TI - Characterization of exposure to extremely low frequency magnetic fields using multidimensional analysis techniques. AB - Our lack of knowledge about the biological mechanisms of 50 Hz magnetic fields makes it hard to improve exposure assessment. To provide better information about these exposure measures, we use multidimensional analysis techniques to examine the relations between different exposure metrics for a group of subjects. We used a combination of a two stage Principal Component Analysis (PCA) followed by an ascending hierarchical classification (AHC) to identify a set of measures that would capture the characteristics of the total exposure. This analysis gives an indication of the aspects of the exposure that are important to capture to get a complete picture of the magnetic field environment. We calculated 44 metrics of exposure measures from 16 exposed EDF employees and 15 control subjects, containing approximately 20,000 recordings of magnetic field measurements, taken every 30 s for 7 days with an EMDEX II dosimeter. These metrics included parameters used routinely or occasionally and some that were new. To eliminate those that expressed the least variability and that were most highly correlated to one another, we began with an initial Principal Component Analysis (PCA). A second PCA of the remaining 12 metrics enabled us to identify from the foreground 82.7% of the variance: the first component (62.0%) was characterized by central tendency metrics, and the second (20.7%) by dispersion characteristics. We were able to use AHC to divide the entire sample (of individuals) into four groups according to the axes that emerged from the PCA. Finally, discriminant analysis tested the discriminant power of the variables in the exposed/control classification as well as those from the AHC classification. The first showed that two subjects had been incorrectly classified, while no classification error was observed in the second. This exploratory study underscores the need to improve exposure measures by using at least two dimensions: intensity and dispersion. It also indicates the usefulness of constructing a typology of magnetic field exposures. PMID- 15832339 TI - Activation of an energy providing response in human keratinocytes after gamma irradiation. AB - We performed a microarray study on human differentiated HaCaT keratinocytes exposed to ionizing radiation (2 or 10 Gy). At 3 h after exposure, more than 150 known and unknown genes were found regulated in irradiated HaCaT keratinocytes. Among the genes regulated at 3 h, those involved in cell energy metabolism appeared to be the most abundant and the most responsive. Two mitochondrial ATP synthases and several other genes involved in energy producing pathways, such as glucose metabolism, were induced, whereas many genes from energy requiring pathways were shut down. These changes in energy metabolism were confirmed both in normal primary keratinocytes and in HaCaT keratinocytes by RT-PCR and proteins studies. Moreover, measures of intracellular ATP revealed a 50% increase in keratinocytes immediately after irradiation, supporting an energy procurement response. The overall results indicate that irradiation induces an immediate burst of ATP that seems to be a general response of human differentiated keratinocytes to the radiation stress. This article contains Supplementary Material available at http://www.mrw.interscience.wiley.com/suppmat/0730 2312/suppmat/v95.html PMID- 15832338 TI - Frequency and amplitude windows in the combined action of DC and low frequency AC magnetic fields on ion thermal motion in a macromolecule: theoretical analysis. AB - We solved the differential equation describing combined action of DC and AC magnetic fields on thermal motion of ions in a biological macromolecule. The solution showed the occurrence of a new set of resonant peaks for ion oscillations under the influence of magnetic fields. After establishment of steady ion oscillations in the macromolecule interior that is well shielded from the action of small particles of the medium surrounding this molecule, the change in energy of ion thermal motion could be sufficient to alter the conformation state of the macromolecule. On this basis, a diversity of biological phenomena can be explained, including the appearance of the known "frequency" and "amplitude" windows, without any resort to the ideas of participation of cyclotron or parametric resonances in these effects. PMID- 15832340 TI - Effects of exposure to a 1950 MHz radio frequency field on expression of Hsp70 and Hsp27 in human glioma cells. AB - Human glioma MO54 cells were used to investigate whether radio frequency (RF) field exposure could activate stress response genes. Cells were exposed to continuous wave 1950 MHz or sham conditions for up to 2 h. Specific absorption rates (SARs) were 1, 2, and 10 W/kg. For the cell growth experiment, cell numbers were counted at 0-4 days after exposure. Expression of Hsp27 and Hsp70, as well as the level of phosphorylated Hsp27 (78Ser) protein, was determined by Western blotting. It was found that sham exposed and RF exposed cells demonstrated a similar growth pattern up to 4 days after RF field exposure. RF field exposure at both 2 and 10 W/kg did not affect the growth of MO54 cells. In addition, there were no significant differences in protein expression of Hsp27 and Hsp70 between sham exposed and RF exposed cells at a SAR of 1, 2, or 10 W/kg for 1 and 2 h. However, exposure to RF field at a SAR of 10 W/kg for 1 and 2 h decreased the protein level of phosphorylated Hsp27 (78Ser) significantly. Our results suggest that although exposure to a 1950 MHz RF field has no effect on cell proliferation and expression of Hsp 27 and Hsp70, it may inhibit the phosphorylation of Hsp27 at Serine 78 in MO54 cells. PMID- 15832341 TI - Breaking and making of the nuclear envelope. AB - During mitosis, a single nucleus gives rise to two nuclei that are identical to the parent nucleus. Mitosis consists of a continuous sequence of events that must be carried out once and only once. Two such important events are the disassembly of the nuclear envelope (NE) during the first stages of mitosis, and its accurate reassembly during the last stages of mitosis. NE breakdown (NEBD) is initiated when maturation-promoting factor (MPF) enters the nucleus and starts phosphorylating nuclear pore complexes (NPCs) and nuclear lamina proteins, followed by NPC and lamina breakdown. Nuclear reassembly starts when nuclear membranes assemble onto the chromatin. This article focuses on the different models of NEBD and reassembly with emphasis on recent data. PMID- 15832343 TI - Great cardiac vein variations. PMID- 15832342 TI - Regulation of chromatin structure during thymic T cell development. AB - Development is the process whereby a multipotent cell gives rise, through series of divisions, to progeny with successively restricted potentials. During T cell development, the process begins with a multipotent hematopoietic stem cell (HSC) in the bone marrow, moves to the thymus where early T cells or thymocytes pass through signal-initiated developmental checkpoints, and ends in the periphery where mature T cells reside. At each step along this developmental pathway, T lymphocyte progenitors must be able to turn genes on and off, creating a specialized program of gene expression, to allow further development. How is gene expression coordinated? This review will summarize what has been learned about the function of chromatin structure in generating a "blueprint" of gene expression during T cell development. This will include discussion of mechanisms of chromatin remodeling, histone modification, and heritable gene silencing. In many cases, these processes are carried out by multi-protein complexes whose components are largely ubiquitously expressed. The spatial and temporal specificity of these complexes is contributed by sequence specific DNA binding factors, some of which are cell type restricted in their expression. This review will summarize research underway to identify these key genetic "targeters." Taken together, the research reviewed here provides a glimpse into the importance of regulation of chromatin structure in T cell development and the "players" involved. PMID- 15832344 TI - JNK/p53 mediated cell death response in K562 exposed to etoposide-ionizing radiation combined treatment. AB - The study of the ability of chemotherapeutic agents and/or ionizing radiation (IR) to induce cell death in tumor cells is essential for setting up new and more efficient therapies against human cancer. Since drug and ionizing radiation resistance is an impediment to successful chemotherapy against cancer, we wanted to check if etoposide/ionizing radiation combined treatment could have a synergic effect to improve cell death in K562, a well-known human erythroleukemia ionizing radiation resistant cell line. In this study, we examined the role played by JNK/SAPK, p53, and mitochondrial pathways in cell death response of K562 cells to etoposide and IR treatment. Our results let us suppose that the induction of cell death, already evident in 15 Gy exposed cells, mainly in 15 Gy plus etoposide, may be mediated by JNK/SAPK pathway. Moreover, p53 is a potential substrate for JNK and may act as a JNK target for etoposide and ionizing radiation. Thus further investigation on these and other molecular mechanisms underlying the cell death response following etoposide and ionizing radiation exposure could be useful to overcome resistance mechanisms in tumor cells. PMID- 15832345 TI - Duplication of the radial artery in the radial forearm flap. AB - The forearm radial flap is currently and widely used free flap, based on the branches of the radial artery, and which, additionally, could contain bone, tendons, or cutaneous nerves. Variations of the radial artery were recently encountered and presented during the elevation of such a flap in 32-year-old male, who sustained war gunshot injury of the left hand. This is the first reported case of a double radial artery in a radial forearm flap in our country, which successfully survived on the recipient hand, although its surface was at the lower threshold for the application of a vascular graft. In rising of the radial forearm flap, pre- and postoperative evaluation of arteries is advisable. PMID- 15832346 TI - Transverse carpal muscle in association with carpal tunnel syndrome: report of three cases. AB - Anomalous muscles of the upper extremity are common, however, symptomatic anomalies causing CTS are rare. Three cases of CTS that are believed to be caused by an anomalous muscle located palmar to the transverse carpal ligament with transversely oriented muscle bundles is presented. Despite the arguments in literature, this is certainly an anomalous muscle that can be encountered during carpal tunnel release and be problematic to manipulate when minimally invasive approaches are chosen. PMID- 15832347 TI - Reciprocal peer teaching: students teaching students in the gross anatomy laboratory. AB - Three common instructional strategies used to teach gross anatomy are lecture, discovery or inquiry-based learning, and cooperative learning. One form of cooperative learning, called reciprocal peer teaching (RPT), illustrates circumstances where students alternate roles as teacher and student. By assuming the responsibility of teaching their peers, students not only improve their understanding of course content, but also develop communication skills, teamwork, leadership, confidence and respect for peers that are vital to developing professionalism early in their medical careers. Traditionally in our Anatomy department, students dissect the entire body using a standard dissection manual. More non-traditionally, however, we have increased cooperative learning in the dissection laboratory by involving students in a series of supplementary RPT activities. During these exercises, 10% of the class practiced their demonstration with course instructors until the students felt prepared to demonstrate the exercise to their classmates. We designed one peer demonstration emphasizing three to six teaching objectives for most of the 40 dissection units. This resulted in a compendium of peer demonstrations for implementation throughout the course. The multitude of diverse exercises permitted each student many opportunities to teach their peers. A debriefing questionnaire was administered at the end of the course demonstrating that 100% of students agreed the RPT experience increased their understanding of the topics they taught and 97% agreed it increased their retention of information they taught to their peers. In addition, 92% agreed that RPT improved their communication skills, which can be applied beyond anatomy to their careers as future physicians. PMID- 15832349 TI - Plantar pressure distribution analysis in normal weight young women and men with normal and claw feet: a cross-sectional study. AB - We analyzed the plantar support in 72 normal-weight young voluntaries (46 women, 26 men), by a baropodometric platform. We considered subjects with claw foot (CFS) and subjects with normal foot (NFS). We found a significant reduction of total plantar support surface in the CFS (P < 0.0001 for women, P < 0.001 for men), due to the reduction of the forefoot and rear foot areas of both plantar imprints. Indeed, CFS of both sexes exhibited higher values of both plantar pressure and peak pressure, compared to the NFS. Moreover, the load per units of plantar surface increased in CFS compared to the NFS. In conclusion, the reduction of plantar support surfaces in CFS of both sexes was associated to a major load per units of plantar surface in the forefoot and rear foot areas, and this may be a risk factor to lower extremity overuse injuries. PMID- 15832348 TI - Unusual attachment of the pectoralis minor muscle. AB - The authors report a presumably unusual bony attachment of the pectoralis minor muscle in an adult cadaver. The specimen's left pectoralis minor had no attachment to the coracoid process of the scapula but attached directly to the fibrous capsule of the glenohumeral joint. Some have theorized that the coracohumeral ligament represents fibers of the pectoralis minor that attach to this bone in some animals but that has degenerated in man. This case report seems to support this possibility. PMID- 15832350 TI - Vortex veins: anatomic investigations on human eyes. AB - The aim of this study was to determine number of ocular vortex veins, their scleral coordinates, and their relationship with nearby extraocular muscles. Sixty intact cadaver orbits having no history of eye or orbital disorders during life were carefully dissected under stereomicroscopic magnification to expose vortex veins and their exit sites from the eyeball. The number of vortex veins per eye varied from four to eight. Eyes having four (35%) or five (30%) vortex veins were observed most frequently. Three eyes (5%) had eight vortex veins. Although the incidence of the vortex veins was variable, there was at least one vein in each quadrant of the sclera. Knowledge of the approximate location of the vortex vein exit sites is very important for surgeons because damage to these veins during eye surgery could produce potential complications, especially choroidal detachment. PMID- 15832351 TI - Structural parameters of the vastus medialis muscle. AB - This research was designed to evaluate musculoskeletal anatomy of the quadriceps region relative to the patellofemoral joint. The hypothesis for the study was that the oblique portion (VMO) of the vastus medialis muscle (VM) is anatomically positioned to function primarily as an active medial stabilizer of the patella. Because many clinicians believe that the VMO functions independently as an active medial stabilizer of the patellofemoral joint (PFJ), PFJ rehabilitation protocols commonly target the VMO in an attempt to restore normal joint mechanics. It is unclear whether this purported selective function is supported by the underlying anatomical structure. Through dissection of 32 limbs from 24 intact cadavers with normal patellar alignment, data were collected on VM fiber alignment and innervation, the presence of fascial plane, and the length of VM about the patella. Statistical analyses demonstrated that the oblique and long heads of the VM muscle had significantly different (P < 0.05) angles of fiber orientation, as expected. When measurements were taken relative to a vertical axis (standardizing limb alignment between cadavers), the difference in fiber angles between oblique and long heads of the VM was reduced significantly. Additionally, < 10% of the length of the VM muscle inserted directly on the medial aspect of the patella, and there was no anatomical evidence of a fascial plane or separate innervation for the oblique and long heads of the VM. The results of the study did not support the hypothesis that the VMO is anatomically positioned to function primarily as an active medial stabilizer of the patella. PMID- 15832352 TI - Anatomic pattern of the terminal branches of posterior interosseous nerve. AB - To understand the lesion of the terminal branches of posterior interosseous nerve (PIN), an anatomic study was carried out. Thirty adult cadaver arms were dissected and the anatomic pattern of the nerve was documented. The distance between the point at which the nerve divides into branches and the distal edge of the supinator were measured, as well as the length of each nerve branch to its muscle-entering sites. The number of branches innervating each muscle was recorded. It was found that the PIN was branching to multiple short branches and a single deep long branch after leaving the supinator muscle with great morphometric and schematic variances. PMID- 15832353 TI - Morphometric study of the human fetal heart. I. Arterial segment. AB - This study measured the dimensions of the great arteries of normal human fetal hearts at an early fetal stage (between 13-20 weeks post-fertilization) in 103 fetuses obtained by necropsy. Different segments of the aorta and the pulmonary artery were dissected and their external diameters measured under stereoscopic magnification. All segments showed linear growth during this period of fetal development. Ranges in median values of external diameters associated with the pulmonary artery were: 2.1-4.2 mm for the valve ring; 2.2-4.2 mm for the main pulmonary trunk; 1.2-2.5 mm for the right pulmonary artery; 0.9-2.18 mm for the left pulmonary artery; and 4.0-8.0 mm for the length of the main pulmonary trunk. Similarly, in the aorta, median diameters were: 2.1-4.2 mm for the ascending aorta; 1.92-3.8 mm for the aortic arch; 1.45-3.0 mm for the aortic isthmus; and 1.75 -3.35 mm for the descending aorta. The diameter of the ductus arteriosus ranged between 1.2-2.45 mm. The growth rates of the pulmonary artery and the aorta were similar. The diameter of the ascending aorta was found to be greater than that of the descending aorta and the right pulmonary artery was wider than the left pulmonary artery. In addition, the magnitude of growth in the various aortic segments was different and the ratios obtained between the aortic isthmus and the ascending and descending aortae, ranged between 0.66-0.93. This study provides important morphometric reference information concerning the dimensions and growth of the great arteries of the fetal heart and has clinical application in pediatric cardiac surgery and echocardiography. PMID- 15832354 TI - Anatomical variations as potential risk factors for ulnar tunnel syndrome: a cadaveric study. AB - The aim of this study was to assess the anatomical variations, especially the anomalous muscles passing through Guyon's canal and the fibrous arch forming the piso-hamate hiatus, which may play a role in ulnar tunnel syndrome. We have also focused on the relation of these structures with specific concern to the ulnar nerve. Nineteen embalmed cadavers (37 hands and forearms) were dissected. A fibrous arch extending between the pisiform and the hook of the hamate was observed in 21 hands. In majority of the cases flexor digiti minimi muscle was found to originate only from this arch. An anomalous muscle was disclosed in six hands with four of them passing through the piso-hamate hiatus with the deep branch of the ulnar nerve. In two of four cases, the superficial branch of the ulnar nerve was also accompanying the deep branch of the ulnar nerve beneath the anomalous muscle and through the piso-hamate hiatus. Because these anomalous muscles were generally found to course through the piso-hamate hiatus with the branches of the ulnar nerve, we conclude that the distal portion of the Guyon's canal has a relatively higher risk for ulnar nerve entrapment. We believe that surgeons operating on this region should take into account these various anatomic structures. PMID- 15832358 TI - Autopsy findings of a 37-year-old man with a complex mosaic karyotype involving del(18p), monosomy 13, and trisomy 20. AB - We report on the autopsy findings of a 37-year-old man with a complex karyotype (mos46,XY,del(18)(p11.1)[14]/46,XY, -13, del (18)(p11.1), +20[8]/47,XY,del(18)(p11.1), +20[8]). He was known to be blind, non-ambulatory, have severe mental retardation, and a seizure disorder. External physical findings at the time of autopsy included micrognathia, short stubby fingers, and rocker bottom feet. Left lobe dominance of the liver and mislocation of the ileocecal junction and appendix were noted on internal examination. The brain was small (700 g) and poorly developed. Microscopically it showed an absence of neurons in the olivary and dentate nuclei, absence of Purkinje cells in the cerebellum, severe depletion of internal granular cells in the cerebellum, and cerebellar dysplasia. Fat infiltration was noted in an unusual distribution in several organs including a pattern in the heart consistent with arrythmogenic right ventricular dysplasia (ARVD). Findings of this mosaic chromosomal karyotype have not been previously described. This report will discuss this individuals physical findings and their relation to similar monochromosomal aberrations. PMID- 15832355 TI - Cystic fibrosis carriers have higher neonatal immunoreactive trypsinogen values than non-carriers. AB - Following cystic fibrosis (CF) neonatal screening implementation, a high frequency of heterozygotes has been reported among neonates with elevated immunoreactive trypsinogen (IRT) and normal sweat chloride levels. We studied the relationship between normal IRT values and CF heterozygosity: 10,000 neonates were screened for CF by IRT measurement and tested for 40 CF mutations; the 294 carriers detected were coupled with newborns negative to the same genetic testing, and the two groups' IRT levels compared. Heterozygotes had higher IRT levels than their controls (mean 35.32 vs. 27.58 microg/L, P<0.001). Even within normal trypsinogen range, the probability of being a CF carrier increases with neonatal IRT concentration. PMID- 15832357 TI - GJB2 (connexin 26) mutations are not a major cause of hearing loss in the Indonesian population. AB - Although hereditary hearing loss is a very heterogeneous disorder, variants in one gene, GJB2 (connexin 26), account for up to 50% of autosomal recessive nonsyndromal sensorineural hearing loss in most populations. This study investigates the contribution of GJB2 to autosomal recessive nonsyndromal hearing loss in the Indonesian population. We performed DNA sequence analysis in 120 patients with profound early childhood nonsyndromal hearing loss and in 100 control individuals and identified three novel variations resulting in amino acid substitutions (p.Gly4Asp, p.Thr5Ala, and p.Gly160Arg). Although we proved that p.Gly4Asp was not disease-causing, the pathological nature of p.Thr5Ala and p.Gly160Arg could not be determined. No recurrent disease-causing mutation could be detected in this Indonesian population. These findings are in contrast with the results obtained in other populations where GJB2 is a major cause of congenital recessive hearing loss. PMID- 15832359 TI - Congenital anomaly of cervical vertebrae is a major complication of Rubinstein Taybi syndrome. AB - Rubinstein-Taybi syndrome (RTS; MIM# 180849) is a well-known malformation syndrome, characterized by broad thumbs and halluces, a characteristic facies, short stature, and mental retardation. RTS is accompanied by a variety of morbid complications, particularly of the skeleton. Based on the experience of five RTS patients with malformation of the craniovertebral junction, we draw attention to previously unrecognized life-threatening complications of RTS, including instability of C1-C2, os odontoideum, hypoplasia of the dens, and fusion of the cervical vertebrae. One patient developed severe cervical myelopathy. Malformation of the cervical spine may be a common syndromic constituent of RTS, to which special attention should be paid to prevent its neurologic sequelae. PMID- 15832360 TI - Adams-Oliver syndrome and hepatoportal sclerosis: occasional association or common mechanism? AB - Adams-Oliver syndrome (AOS) is characterized by the association of scalp and skull defects and abnormalities of terminal limbs. Congenital heart malformations have also been reported. Hepatoportal sclerosis (HPS) is a rare cause of portal hypertension in children characterized by abnormalities of intra-hepatic portal veins, portal fibrosis, and nodular regeneration. Etiopathogenesis of these rare disorders remains unclear, but the hypothesis of vascular thrombotic mechanism has been suggested. Association of both syndromes has been reported in only one child. We now report on two unrelated children with AOS and HPS, one child harboring a factor V Leiden mutation. We hypothesize that the association of both disorders may not be fortuitous and reinforces the idea that AOS and HPS may share a vascular thrombotic mechanism. PMID- 15832356 TI - Spectrum of malformations of the hindbrain (cerebellum, pons, and medulla) in a cohort of children with high rate of parental consanguinity. AB - We review 25 patients with a spectrum of hindbrain (cerebellum, pons, and medulla) malformations from a cohort of children with high parental consanguinity rate. Twenty-three of the 25 patients were born to consanguineous parents. The patients were classified in four groups. Eleven patients of 6 families had malformation of the hindbrain and midbrain with molar tooth sign (10 patients of 5 families with typical Joubert syndrome), 5 patients showed severe supratentorial anomalies in addition to the hindbrain malformations, 5 patients had pontocerebellar or cerebellar hypoplasia with anterior horn cell disease in the spinal cord (spinal muscular atrophy), and 4 patients showed malformations affecting predominantly the hindbrain without substantial involvement of other systems. A locus for Joubert syndrome was previously identified on chromosome 9q34.3 in two families, and a second locus on chromosome 11p12-q13.3 in another family. A third Joubert syndrome locus has been mapped at 6q23 and a mutation in the AHI1 gene at this site has been found recently in a further family from this cohort. Delineation of homogeneous subgroups of patients with hindbrain malformations and molecular genetic analysis of these groups may lead to identification of further loci, genes and mutations responsible for the malformations. PMID- 15832362 TI - Monozygotic twins concordant for blood karyotype, but phenotypically discordant: a case of "mosaic chimerism". AB - We report on 23 years old discordant monozygotic (MZ) twins, one with minor anomalies and mental delay, the other one being normal. Both had 46,XX,dup(11)(p12p15)/46,XX mosaicism in blood, with a similar proportion of abnormal cells (respectively, 16% and 17%). However, interphase fluorescence in situ hybridization (FISH) analysis performed on buccal smear and urinary sediment using specific probes located at the duplicated region showed that mosaicism was only present in the abnormal twin, with 68% abnormal cells. We hypothesize that the postzygotic chromosomal rearrangement may have occurred early in one embryo after the twinning event, and the blood mosaicism observed in both twins would have resulted from blood exchanges via placental anastomoses. This hypothesis of chimerism is strongly supported by twin-to-twin transfusion syndrome observed during fetal life of our twins. This case and those previously reported lead us to suggest that blood is particularly unsuitable for cytogenetic investigations of twins. PMID- 15832363 TI - Identification and characterization of protein subcomplexes in yeast. AB - Protein complexes are major components of cellular organization. Based on large scale protein complex data, we present the first statistical procedure to find insightful substructures in protein complexes: we identify protein subcomplexes (SCs), i.e., multiprotein assemblies residing in different protein complexes. Four protein complex datasets with different origins and variable reliability are separately analyzed. Our method identifies well-characterized protein assemblies with known functions, thereby confirming the utility of the procedure. In addition, we also identify hitherto unknown functional entities consisting of either functionally unknown proteins or proteins with different functional annotation. We show that SCs represent more reliable protein assemblies than the original complexes. Finally, we demonstrate unique properties of subcomplex proteins that underline the distinct roles of SCs: (i) SCs are functionally and spatially more homogeneous than complete protein complexes (this fact is utilized to predict functional roles and subcellular localizations for so far unannotated proteins); (ii) the abundance of subcomplex proteins is less variable than the abundance of other proteins; (iii) SCs are enriched with essential and synthetic lethal proteins; and (iv) mutations in SC-proteins have higher fitness effects than mutations in other proteins. PMID- 15832364 TI - Structural determination of N-linked glycans by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry. AB - This paper reviews methods for the analysis of N-linked glycans by mass spectrometry with emphasis on studies conducted at the Oxford Glycobiology Institute. Topics covered are the release of glycans from sodium dodecyl sulphate polyacrylamide gel electrophoresis gels, their purification for analysis by mass spectrometry, methods based on matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization for producing fragment ions, and details of their fragmentation. MALDI mass spectrometry provided a rapid method for profiling neutral N-linked glycans as their [M + Na](+) ions which could be fragmented by collision-induced decomposition to give spectra containing both glycosidic and cross-ring fragments. Electrospray ionization mass spectrometry was more versatile in that it was relatively easy to change the type of ion that was formed and, furthermore, unlike MALDI, electrospray did not cause extensive loss of sialic acids from sialylated glycans. Negative ions formed by addition of anions such as chloride and, particularly, nitrate, to the electrospray solvent were stable and enabled singly charged ions to be obtained from larger glycans than was possible in positive ion mode. Fragmentation of negative ions followed specific pathways that defined structural details of the glycans that were difficult to obtain by classical methods such as exoglycosidase digestion. PMID- 15832361 TI - Transforming growth factor-beta1 (TGF-beta1) regulates ATDC5 chondrogenic differentiation and fibronectin isoform expression. AB - Regulated splicing of fibronectin (FN) occurs during the mesenchymal to chondrocyte transition and ultimately results in the relative enrichment of an extra domain B (EDB) exon-containing FN isoform with the suggestion that FN isoforms may play a functional role in chondrogenesis. Promotion of chondrogenesis can also be achieved by treatment with transforming growth factor beta (TGF-beta), which also regulates FN isoform expression. We have examined the effects of TGF-beta treatment on the assumption of the chondrogenic phenotype in the teratoma-derived cell line ATDC5 and tested whether these effects on chondrogenesis are paralleled by appropriate changes in FN isoform expression. ATDC5 cells were maintained in a pre-chondrogenic state and, in this state, treated with 10 ng/ml TGF-beta. The cells started to elaborate a matrix rich in sulfated proteoglycans, such that within the first 12 days of culture, TGF-beta1 treatment appeared to slightly accelerate early acquisition of an Alcian blue stained matrix, and caused a dose- and time-dependent decrease in collagen type I expression; changes in collagen type II expression were variable. At later times, cells treated with TGF-beta became indistinguishable from those of the controls. Interestingly, TGF-beta treatment caused a significant dose- and time-dependent decrease in the proportion of FN containing the extra domain A (EDA) and the EDB exons. These data suggest that TGF-beta induces the early stages of chondrogenic maturation in this pre-chondrogenic line and that TGF-beta treatment increases expression of FN isoforms that lack the EDA and EDB exons. PMID- 15832365 TI - Comparative proteomic analysis of peripheral blood eosinophils from healthy donors and atopic dermatitis patients with eosinophilia. AB - Atopic dermatitis (AD) is an allergic disease that has recently shown a dramatic increase of incidence in developed countries. Eosinophilia, the accumulation of eosinophils, occurs in AD patients through an anti-apoptotic mechanism. To understand the target proteins involved in the anti-apoptotic signaling of eosinophilia, we used a proteomic approach to analyze eosinophil proteins from AD patients with eosinophilia and healthy donors. Protein spots in two-dimensional electrophoresis (2-DE) gels were identified with peptide mass fingerprinting (PMF) based on matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and database searching. More spots were observed in the 2-DE proteome map from AD patient samples (1310 +/- 58 spots) than in those from healthy donors (1121 +/- 40 spots). We identified 51 proteins affected by eosinophilia: 19 related to signaling, 8 involved in regulation of metabolism, 4 related to apoptosis, and 3 involved in inflammation. The other identified proteins were associated with transcription, RNA processing, translation, the cytoskeleton, and unknown functions. Among the identified proteins, we observed prominent increases in the expressions of cyclinA2, voltage-dependent anion channel protein 2, and 38 kDa FK506 binding protein 8 in eosinophils from AD patients in comparison to healthy donors. PMF and immunoblotting of a single spot that was expressed in eosinophils from healthy individuals but not in AD patients identified the protein as phosphorylated growth receptor binding 7 (Grb7) adaptor protein. Increased phosphorylation of Grb7 and its upstream signaling protein, focal adhesion kinase (FAK), was detected in low viability eosinophils such as those from healthy donors or in cultured eosinophils (AML14.3D10 cells) treated with dexamethasone. These results suggest that phosphorylation of Grb7 and the expressions of cyclinA2, voltage-dependent anion channel protein 2, and 38 kDa FK506 binding protein 8 may be related with the anti-apoptosis mechanism of eosinophilia. PMID- 15832366 TI - Peptide microarrays for the detection of molecular interactions in cellular signal transduction. AB - The formation of protein complexes is a hallmark of cellular signal transduction. Here, we show that peptide microarrays provide a robust and quantitative means to detect signalling-dependent changes of molecular interactions. Recruitment of a protein into a complex upon stimulation of a cell leads to the masking of an otherwise exposed binding site. In cell lysates this masking can be detected by reduced binding to a microarray carrying a peptide that corresponds to the binding motif of the respective interaction domain. The method is exemplified for the lymphocyte-specific tyrosine kinase 70 kDa zeta-associated protein binding to a bis-phosphotyrosine-motif of the activated T-cell receptor via its tandem SH2 domain. Compared to established techniques, the method provides a significant shortcut to the detection of molecular interactions. PMID- 15832369 TI - Towards a proteomic definition of CoArtem action in Plasmodium falciparum malaria. AB - We have adopted a proteomic strategy to investigate the actions of the two active components of the new antimalarial CoArtem, artemether and lumefantrine, following pharmacologically relevant drug exposure in the human malaria parasite Plasmodium falciparum. Both drugs induced profound alterations in the parasite's proteome. Moreover, the pattern of proteome alteration was specific for the drug used. The two drugs induced opposing effects on key glycolytic enzymes while exerting similar influence of the expression of stress response proteins. These initial results demonstrate the power of this approach in the study of pleiomorphic mechanisms of drug action. PMID- 15832368 TI - A proteome analysis of the yeast response to the herbicide 2,4 dichlorophenoxyacetic acid. AB - The intensive use of herbicides may give rise to a number of toxicological problems in non-target organisms and has led to the emergence of resistant weeds. To gain insights into the mechanisms of adaptation to the herbicide 2,4 dichlorophenoxyacetic acid (2,4-D), we have identified variations in protein expression level in the eukaryotic experimental model Saccharomyces cerevisiae exposed to herbicide aggression, based on two-dimensional gel electrophoresis. We show results suggesting that during the adaptation period preceding the resumption of inhibited exponential growth under herbicide stress, the antioxidant enzyme Ahp1p and the heat shock proteins Hsp12p and Ssb2p (or Ssb1p) are present in higher amounts. The increased level of other enzymes involved in protein (Cdc48p) and mRNA (Dcp1p) degradation, in carbohydrate metabolism (Eno1p, Eno2p and Glk1p) and in vacuolar H(+)-ATPase (V-ATPase) function (Vma1p and Vma2p, two subunits of the peripheral catalytic sector) was also registered. V ATPase is involved in the homeostasis of intracellular pH and in the compartmentalization of amino acids and other metabolites in the vacuole. The increased expression of amino acid biosynthetic enzymes (Arg1p, Aro3p, Aro8p, Gdh1p, His4p, Ilv3p and Met6p), also suggested by comparative analysis of the proteome, was correlated with the reduction of amino acid concentration registered in both the vacuole and the cytosol of 2,4-D-stressed cells, possibly due to the disturbance of vacuolar and plasma membrane functions by the lipophilic acid herbicide. PMID- 15832367 TI - Proteomic identification of Ku70/Ku80 autoantigen recognized by monoclonal antibody against hepatocellular carcinoma. AB - A murine monoclonal antibody (mAb), CLD3 (IgG(1),kappa), was generated against hepatocellular carcinoma (HCC). Both immunofluorescence and immunohistochemical assays indicated the reactivity of CLD3 mAb localized at the nucleus and/or cytoplasm of tumorigenic HCC cell lines as well as in liver cancer tissues. By immunoprecipitation and using the matrix-assisted laser desorption/ionization time of flight mass spectrometry approach, the antigenic specificity of CLD3 was determined to be heterodimeric Ku70 and Ku80 autoantigen, which was confirmed by Western blotting. PMID- 15832371 TI - Increased expression of calcineurin in human colorectal adenocarcinomas. AB - Colorectal cancer (CRC) is the third most common cause of cancer death in the Western world. Calcineurin (CaN), a Ca2+/calmodulin (CaM)-dependent protein phosphatase, is important for Ca2+-mediated signal transduction. The main objective of this study is to examine the potential role of Ca2+/CaM-dependent protein phosphatase in both normal and in invasive tumor components of human samples. In this study, we carried out 45 cases of CaN activity, 13 cases of CaN protein expression by Western blot analysis, and 6 cases for immunohistochemical analysis in both normal and invasive tumor components of human samples. Immunohistochemical analysis revealed that strong cytoplasmic staining of varying intensity was observed in colon tumors of all patients compared to normal mucosa. In addition, Western blot analysis revealed a prominent overexpressed immunoreactive band with an apparent molecular mass of 60 kDa catalytic alpha subunit (CaN A) as well as CaN Aalpha and beta in colon tumor samples. Elevated CaN protein expression appears to be a possible link between Ca2+ signaling and oncogenic processes. PMID- 15832370 TI - A rare and often unrecognized cerebromeningitis and hemodynamic disorder: a major cause of sudden death in somatic cell cloned piglets. AB - In this study, we generated 40 somatic cell cloned (scNT) piglets. Of these, five piglets were stillborn, 22 scNT piglets died suddenly within the first week of life, and 1 piglet died after 40 days. Twelve scNT piglets are still healthy. The birth weights of compromised scNT piglets in comparison with those of normal scNT piglets are significantly reduced (0.80 +/- 0.29 vs 1.27 +/- 0.30 kg, p < 0.05), in spite of longer gestation (114 versus 120 day). Significant findings from histological examinations showed that approximately 25% (7/28) of scNT piglets showed severe congestion of lung and liver or neutrophilic inflammation in brain indicating that unexpected phenotypes can appear as a result of somatic cell cloning. Two-dimensional gel electrophoresis experiments revealed changes in the responses of several detoxification-related proteins related to stress and inflammation and found significant alterations in myocardium-specific proteins, indicating hemodynamic disorder. scNT piglets that survived to adulthood did not show any abnormality except skin and hair color depigmentation. The present study suggests that cerebromeningitis and hemodynamic disorder are a major risk factor for sudden early death of scNT piglets. Although we cannot completely exclude the possibility that scNT piglets are susceptible to specific respiratory infections, our data suggests that the early death of scNT clones is due to cardiopulmonary functional abnormalities and cerebromeningitis. PMID- 15832373 TI - Production of soluble matriptase by human cancer cell lines and cell surface activation of its zymogen by trypsin. AB - The membrane-bound serine proteinase matriptase, which is often released from the plasma membrane of epithelial and carcinoma cells, has been implicated to play important roles in both physiological and pathological conditions. However, the regulatory mechanism of its activity is poorly understood. In the present study, we examined expression and activation state of soluble matriptase in 24 human cancer cell lines. Soluble matriptase was detected in the conditioned media from all of 5 colon and 4 breast carcinoma cell lines and 8 of 10 stomach carcinoma cell lines tested. Only two of five lung cancer cell lines released the matriptase protein into the culture media. Out of the five matriptase-negative cell lines, two cell lines expressed the matriptase mRNA. Among 24 cancer cell lines tested, 13 cell lines secreted trypsin in an active or latent form and all of them released matriptase. Most of the 24 cell lines released a latent, single chain matriptase of 75 kDa as a major form, as well as low levels of complex forms of an activated two-chain enzyme with its specific inhibitor HAI-1. Thus, these soluble matriptases appeared to have little proteolytic activity. Treatment of stomach and colon cancer cell lines with epidermal growth factor stimulated the release of matripatase/HAI-1 complexes. In cancer cell lines secreting active trypsin, however, matriptase was released mostly as an inhibitor-free, two-chain active form. Trypsin seemed to activate the membrane-bound, latent matriptase on the cell surface. These results suggest that matriptase and trypsin cooperatively function for extracellular proteolysis. PMID- 15832372 TI - Biological evaluation of penetration domain and killing domain peptides. AB - BACKGROUND: Cancer gene therapy must impact the majority of cells to be effective. Current gene delivery systems are unable to achieve sufficient transfer efficiency to the tumor cells. Cell killing can be dramatically increased through a bystander effect. Modeling the gene product with synthetic peptides can identify key elements for creating cell killing through a bystander effect. METHODS: Fluorescent labeled peptides were used for uptake kinetic studies and determination of intracellular localization in human glioblastoma cell lines, rat glioma cells lines and pressurized rat cerebral arteries. The degree of cell killing was assayed using propidium iodide coupled with fluorescence-activated cell sorting (FACS) analysis. RESULTS: Peptides derived from HIV Tat and Drosophila antennapedia homeodomain were taken up by all tumor and primary cells. Attachment of an Mdm-2-binding domain derived from P14(ARF) resulted in cell killing and was independent of domain orientation. Uptake kinetics showed rapid uptake for both tumor and primary cells equilibrating with the external media within 10 min. Intraluminal or extraluminal administration of peptides into pressurized cerebral arteries showed a lack of extravasation across the subbasement lamina. Assay of biological activity following intraluminal administration showed selective suppression of response to vasodilation with no effect on response by smooth muscle cells. CONCLUSIONS: The results from these studies identified: (1) a cell trafficking domain and a cytotoxic domain for killing brain tumor cells; (2) that cell killing was independent of the domain orientations with regard to the cell trafficking domain being at the C-terminus or N-terminus; and (3) that the dual domain peptide can also be taken up by endothelial cells as shown by the cerebral artery studies. Hence, localized expression of the cytotoxic gene has the potential to not only kill brain tumor cells, but also tumor endothelium, thus further increasing the effectiveness of the therapy. PMID- 15832374 TI - Site-directed mutations (Asp405Ile and Glu124Ile) in cytochrome P450scc: effect on adrenodoxin binding. AB - Cytochrome P450scc, mitochondrial adrenodoxin (Adx), and adrenodoxin reductase (AdR) are an essential components in a steroid hydroxylation system. In particular, mytochondrial cytochrome P450scc enzyme catalyses the first step in steroid hormones biosynthesis, represented by the conversion of cholesterol to pregnenolone. In order to study the effect of single mutations on the Adx binding a model of bovine cytochrome P450scc, previously optimized by molecular modeling, was utilized. It was hypothesized by molecular docking that two residues (Asp405 and Glu124) are involved in Adx binding. By site-directed mutagenesis, two mutants of cytochrome P450scc (Asp405Ile and Glu124Ile) expressed in Escherichia coli, were realized by replacing with isoleucines. The site-directed mutations effect on Adx binding was evaluated by differential spectral titration. The apparent dissociation constant values for Asp405Ile and Glu124Ile cytochrome P450scc show that the mutated residues seem to be at the interaction domain with Adx or at least close to it, as predicted by molecular modeling study. Finally, the engineered enzymes were characterized by biochemical and biophysical techniques such as circular dichroism (CD), UV/Vis spectroscopy, and electrochemical analysis. PMID- 15832375 TI - Suicidal genetically engineered microorganisms for bioremediation: need and perspectives. AB - In the past few decades, increased awareness of environmental pollution has led to the exploitation of microbial metabolic potential in the construction of several genetically engineered microorganisms (GEMs) for bioremediation purposes. At the same time, environmental concerns and regulatory constraints have limited the in situ application of GEMs, the ultimate objective behind their development. In order to address the anticipated risks due to the uncontrolled survival/dispersal of GEMs or recombinant plasmids into the environment, some attempts have been made to construct systems that would contain the released organisms. This article discusses the designing of safer genetically engineered organisms for environmental release with specific emphasis on the use of bacterial plasmid addiction systems to limit their survival thus minimizing the anticipated risk. We also conceptualize a novel strategy to construct "Suicidal Genetically Engineered Microorganisms (SGEMs)" by exploring/combining the knowledge of different plasmid addiction systems (such as antisense RNA-regulated plasmid addiction, proteic plasmid addiction etc.) and inducible degradative operons of bacteria. PMID- 15832377 TI - MHC-I recognition by receptors on myelomonocytic cells: new tricks for old dogs? AB - Receptors on cytotoxic T lymphocytes and natural killer cells play well established roles in the immunological response and share a common ligand in the form of MHC-I. We discuss how a variety of MHC-I receptors are also expressed on myelomonocytic cells such as macrophages and dendritic cells. Since myelomonocytic MHC-I receptors recognise a broad range of alleles and MHC-I structures, we propose that their task is to discern expression levels and folding forms of MHC. We describe a model in which these recognition events would regulate bidirectional cross talk between cells of innate and adaptive immune systems to organise an ongoing combined immune response. We discuss how such a model is supported by recent literature and might function in a variety of contexts, including immunoregulation during pregnancy. Our model also offers an alternative explanation of immune dysregulation rather than autoimmunity during HLA-B27-associated spondyloarthropathies and addresses a number of conundrums in this field. PMID- 15832378 TI - Sperm-mediated gene transfer: applications and implications. AB - Recent developments in studies of sperm-mediated gene transfer (SMGT) now provide solid ground for the notion that sperm cells can act as vectors for exogenous genetic sequences. A substantive body of evidence indicates that SMGT is potentially useable in animal transgenesis, but also suggests that the final fate of the exogenous sequences transferred by sperm is not always predictable. The analysis of SMGT-derived offspring has shown the existence of integrated foreign sequences in some cases, while in others stable modifications of the genome are difficult to detect. The appearance of SMGT-derived modified offspring on the one hand and, on the other hand, the rarity of actual modification of the genome, suggest inheritance as extrachromosomal structures. Several specific factors have been identified that mediate distinct steps in SMGT. Among those, a prominent role is played by an endogenous reverse transcriptase of retrotransposon origin. Mature spermatozoa are naturally protected against the intrusion of foreign nucleic acid molecules; however, particular environmental conditions, such as those occurring during human assisted reproduction, can abolish this protection. The possibility that sperm cells under these conditions carry genetic sequences affecting the integrity or identity of the host genome should be critically considered. These considerations further suggest the possibility that SMGT events may occasionally take place in nature, with profound implications for evolutionary processes. PMID- 15832376 TI - A walk from the wild side: the genetics of domestication of livestock and crops. AB - The phenotypic variation found in domesticated plants and animals is striking, so much so that Darwin used it to illustrate the power of selection to effect change. Recent developments in genomics technologies are leading to dramatic progress in elucidating the genetic changes that occur during domestication. The Genetics Society Autumn Meeting on the genetics of domestication took place in November 2004 at the Royal Society in London, and was organised by Helen Sang (Roslin Institute, UK) and Jonathan Jones (John Innes Centre, UK). The meeting brought together many of the leading researchers on livestock and crop domestication and provided a timely and exciting account of recent progress in the field. PMID- 15832379 TI - Concepts in nuclear architecture. AB - Genomes are defined by their primary sequence. The functional properties of genomes, however, are determined by far more complex mechanisms and depend on multiple layers of regulatory control processes. A key emerging contributor to genome function is the architectural organization of the cell nucleus. The spatial and temporal behavior of genomes and their regulatory proteins are now being recognized as important, yet still poorly understood, control mechanisms in genome function. Combined cell biological, molecular and computational analysis of architectural aspects of genome function has added a further dimension to the investigation of some of the most fundamental cellular processes including transcription and maintenance of genome integrity. The complete elucidation of the contribution that nuclear architecture makes to gene expression will be required to fully understand physiological processes such as differentiation, development and disease at the cellular level. Here I give an overview of some of the emerging concepts in the study of in vivo genome organization and function. PMID- 15832380 TI - Evolutionary innovation in the vertebrate jaw: A derived morphology in anuran tadpoles and its possible developmental origin. AB - The mouthparts of anuran tadpoles are highly derived compared to those of caecilians or salamanders. The suprarostral cartilages support the tadpole's upper beak; the infrarostral cartilages support the lower beak. Both supra- and infrarostral cartilages are absent in other vertebrates. These differences reflect the evolutionary origin of a derived feeding mode in anuran tadpoles. We suggest that these unique cartilages stem from the evolution of new articulations within preexisting cartilages, rather than novel cartilage condensations. We propose testing this hypothesis through a search for similarities in the development of the suprarostral and infrarostral cartilage articulations and of the primary jaw joint. In Xenopus, the gene zax is expressed in a region corresponding to the infrarostral cartilage. This gene is related to the bapx1 gene, which regulates jaw joint development. Further investigation of these genes, as well as other genes with joint-related functions, in anuran craniofacial development may provide a connection between the morphological diversity seen in the vertebrate head and the corresponding diversity in genetic regulatory processes. We believe that the evolution of larval jaws in anurans may shed light on the general evolutionary mechanisms of how new articulations, not only in the jaw region, could have arisen in the vertebrate skull. PMID- 15832381 TI - Patterned cell determination in a plant tissue: the secondary phloem of trees. AB - The secondary vascular tissues (xylem and phloem) of woody plants originate from a vascular cambium and develop as radially oriented files of cells. The secondary phloem is composed of three or four cell types, which are organised into characteristic recurrent cellular sequences within the radial cell files of this tissue. There is a gradient of auxin (indole acetic acid) across both the cambium and the immediately postmitotic cells within the xylem and phloem domains, and it is believed that this morphogen, probably in concert with other morphogenic factors, is closely associated with the determination and differentiation of the different cells types in each tissue. A hypothesis is developed that, in conjunction with the positional values conferred by the graded radial distribution of morphogen, cell divisions at particular positions within the cambium are sufficient to determine not only each of the phloem cell types but also their recurrent pattern of differentiation within each radial cell file. PMID- 15832384 TI - Male sex drive and the masculinization of the genome. AB - Charles Darwin remarked that "males, with their superior strength, pugnacity, armaments, unwieldly passion and love songs, are almost always the more active and most often, the initiators of sexual interactions". Here, we propose that such male sex drive directly impacts the genome by leading to its progressive masculinization--genes that possess sex-specific effects on male fitness accumulate to a much greater extent and are generally more diverged. The larger proportion of male versus female fitness modifiers in combination with stronger sexual selection may generate evolutionary signatures such as a greater sensitivity to male sterility and a paucity of X-linked male-specific genes. Male sex-drive theory complements the female-choice theory of sexual selection and allows for the genetic variation of costly sexual traits to be continuously replenished. PMID- 15832383 TI - A concise peer into the background, initial thoughts and practices of human gene therapy. AB - The concept of human gene therapy came on the heels of fundamental discoveries on the nature and working of the gene. However, realistic prospects to correct the underlying cause of recessive genetic disorders through the transfer of wild-type alleles of defective genes had to wait for the arrival of recombinant DNA technology. These techniques permitted the isolation and insertion of genes into the first recombinant delivery systems. The realization that viruses are natural gene carriers provided inspiration for gene therapy and, as engineered vectors, viruses became prominent gene delivery vehicles. Nonetheless, when put in the context of human and non-human primate studies, all vectors fell short of success regardless of their viral or non-viral origin. Recognition of issues such as inefficient gene transfer and short-lived or scant expression in the relevant cell type(s) prompted researchers to refine and develop several gene delivery systems, in particular those based on retroviruses, adeno-associated viruses and adenoviruses. Concomitantly, available technology was deployed to tackle disorders that require few genetically corrected cells to attain therapy. PMID- 15832382 TI - How and when did Arabidopsis thaliana become highly self-fertilising. AB - Changes in breeding system are a regular evolutionary change in plants, as self fertilisation is often advantageous, particularly for weedy and colonising species. The adoption of Arabidopsis thaliana as a plant model species has led to interest in how self-incompatibility was lost so that this species became highly inbreeding. Molecular evolutionary approaches have recently focused on investigating two loci involved in the incompatibility recognition process in related Arabidopsis species; non-functional copies of these genes still exist in A. thaliana. New work studying polymorphism at these loci found strikingly low diversity at one of them, suggesting that spread of a mutation in this gene might have caused self-compatibility in an ancestor of A. thaliana. However, it is difficult to be sure of the time when the selfing habit evolved in the lineage that led to A. thaliana. PMID- 15832385 TI - The three-sided romance of the lateral line: glia love axons love precursors love glia. AB - The lateral line system of fish and amphibians is closely related to the inner ear in terms of evolution, morphology and physiology. Several recent papers have shed new light on the postembryonic development of this system, and have revealed an unexpected triangular relationship where migrating sensory precursors guide axons, axons guide glia and glia, in turn, control the formation of sensory organs. They have also revealed the crucial importance of controlled cell migration not only for patterning the system, but also for determining polarity (and therefore directional sensitivity) of the mechanosensory hair cells. The remarkable accessibility of the lateral line system may allow a detailed analysis of cell migration and polarization, and may help us better understand the complex interactions between sensory precursor cells, neurons and glia during development. PMID- 15832386 TI - STOP and GO with NO: nitric oxide as a regulator of cell motility in simple brains. AB - During the formation of the brain, neuronal cell migration and neurite extension are controlled by extracellular guidance cues. Here, I discuss experiments showing that the messenger nitric oxide (NO) is an additional regulator of cell motility. NO is a membrane permeant molecule, which activates soluble guanylyl cyclase (sGC) and leads to the formation of cyclic GMP (cGMP) in target cells. The analysis of specific cells types in invertebrate models such as molluscs, insects and the medicinal leech provides insight how NO and cyclic nucleotides affect the wiring of nervous systems by regulating cell and growth-cone motility. Inhibition of the NOS and sGC enzymes combined with rescue experiments show that NO signalling orchestrates neurite outgrowth and filopodial dynamics, cell migration of enteric neurons, glial migration and axonogenesis of pioneer fibers. Cultured insect embryos are accessible model systems in which cellular mechanisms of NO-induced cytoskeletal reorganizations can be analyzed in natural settings. Finally, I will outline some indications that NO may also regulate cell motility in the developing and regenerating vertebrate nervous system. PMID- 15832387 TI - Nonsense-mediated RNA decay: a molecular system micromanaging individual gene activities and suppressing genomic noise. AB - Nonsense-mediated RNA decay (NMD) is an evolutionary conserved system of RNA surveillance that detects and degrades RNA transcripts containing nonsense mutations. Given that these mutations arise at a relatively low frequency, are there any as yet unknown substrates of NMD in a wild-type cell? With this question in mind, Mendell et al. have used a microarray assay to identify those human genes under NMD regulation. Their results show that, in human cells, NMD regulates hundreds of physiologic transcripts and not just those containing nonsense mutations. Among the NMD targets are a number of non-functional RNAs expressed from vestigial sequences derived from retroviral and transposable elements. These findings support the notion that NMD is a high profile post transcriptional mechanism micromanaging the activity of multiple gene batteries and suppressing the expression of genetic remnants. PMID- 15832388 TI - A metabolic enzyme doing double duty as a transcription factor. AB - Many kinds of multifunctional regulatory proteins have been identified that perform distinct biochemical functions in the nucleus, the cytoplasm, or both. Here we describe the recent discovery by Hall et al. (2004) of a new type of multifunctional protein: a metabolic enzyme that doubles as a transcription factor. This enzyme, Arg5,6, functions as a catalytic enzyme in ornithine biosynthesis and also binds and regulates the promoters of nuclear and mitochondrial genes. It may also regulate precursor mRNA metabolism. We discuss how proteins that serve as both metabolic enzymes and transcription factors might have evolved. PMID- 15832390 TI - Luminescent gold(III) alkynyl complexes: synthesis, structural characterization, and luminescence properties. PMID- 15832389 TI - Synthesis of the tetracyclic core of the tetrapetalones through transannular oxidative [4+3] cyclization. PMID- 15832392 TI - Determination by asymmetric total synthesis of the absolute configuration of lucilactaene, a cell-cycle inhibitor in p53-transfected cancer cells. PMID- 15832391 TI - Donor-acceptor pretzelanes and a cyclic bis[2]catenane homologue. PMID- 15832393 TI - Organic-inorganic hybrid sol-gel materials incorporating functionalized cobalt(III) corroles for the selective detection of CO. PMID- 15832394 TI - The myth of cyanide always being a strong-field ligand: synthesis and structural characterization of homoleptic S=2 pentacyanochromate(II), [Cr(II)(CN)5]3-, and nonacyanodichromate(II), [Cr(II)2(CN)9]5-. PMID- 15832395 TI - One-pot sequential N and C arylations: an efficient methodology for the synthesis of trans 4-N,N-diaryl aminostilbenes. PMID- 15832396 TI - Contributions of the distance-dependent reorganization energy and proton-transfer to the hole-transfer process in DNA. AB - A kinetic study of the single-step hole transfer in DNA was performed by measuring time-resolved transient absorption. DNA molecules with various sequences were designed and conjugated with naphthalimide (NI) and phenothiazine (PTZ) to investigate the sequence and distance dependence of the single-step hole transfer between guanines (Gs). Hole injection into DNA was accomplished by excitation of the NI site with a 355 nm laser pulse, and the kinetics of the hole transfer process were investigated by monitoring the transient absorption of the PTZ radical cation (PTZ.+). Kinetic analysis of the time profile of PTZ.+ based on the kinetic model showed that the distance dependence of the hole-transfer process was significantly influenced by the DNA sequence. Results of temperature- and isotope-effect experiments demonstrated that the activation energy increased as the number of bridge bases separating the Gs increased. This is because of the distance-dependent reorganization energy and contribution of the proton-transfer process to the hole transfer in DNA. PMID- 15832397 TI - Complete assignment of heteronuclear protein resonances by protonless NMR spectroscopy. PMID- 15832398 TI - Triple-bond covalent radii. AB - A system of additive covalent radii is proposed for sigma(2) pi(4) triple bonds involving elements from Be to E 112 (eka-mercury). Borderline cases with weak multiple bonding are included. Only the elements in Group 1, the elements Zn-Hg in Group 12 and Ne in Group 18 are then totally excluded. Gaps are left at late actinides and some lanthanides. The standard deviation for the 324 included data points is 3.2 pm. PMID- 15832399 TI - Design, synthesis, and photodegradation of silicon-containing polyureas. AB - Novel N-phenyl aromatic polyureas containing bis[(N,N' diphenylureylene)methyl]silane units in the skeleton were designed as a new type of photodegradable polymer. These materials were successfully prepared in 88-93 % yields by copolymerization of bis(anilinomethyl)dimethylsilane and dianilino-p xylene with 4,4'-methylenebis(phenylisocyanate). Their photodegradability was found to be 10.1 times higher than that of polymers of similar structure, but lacking the silyl unit. Furthermore, the photodegradation mechanism of polyureas was elucidated, and involves single-electron transfer between silyl and carbonyl groups, silyl group migration, and solvolysis. These novel polymers are potential materials of high economic value for use in photolithography and microelectronics. PMID- 15832400 TI - Design of organic semiconductors: tuning the electronic properties of pi conjugated oligothiophenes with the 3,4-ethylenedioxythiophene (EDOT) building block. AB - Hybrid oligothiophenes based on a various combinations of thiophene and 3,4 ethylenedioxythiophene (EDOT) groups have been synthesized. UV/Vis absorption spectra show that the number and relative positions of the EDOT groups considerably affect the width of the HOMO-LUMO gap and the rigidity of the conjugated system. Analysis of the crystallographic structure of two hybrid quaterthiophenes confirms that insertion of two adjacent EDOT units in the middle of the molecule leads to a self-rigidification of the conjugated systems by intramolecular SO interactions. Cyclic voltammetry data shows that the first oxidation potential of the oligomers decreases with increasing chain length and increasing number of EDOT groups for a given chain length. Electrochemical studies and theoretical calculations show that the positions of the EDOT units in the conjugated chain control the potential difference (DeltaE(p)) between the first and second oxidation steps. Moving the EDOT groups from the outer to the inner positions of the conjugated system increases DeltaE(p). Theoretical calculations confirm that this phenomenon reflects an increase of the intramolecular coulombic repulsion between positive charges in the dication. A thin-film field-effect transistor was fabricated by vacuum sublimation of a pentamer with alternating thiophene-EDOT structure, and the hole mobility was determined. PMID- 15832401 TI - Electronic energy levels in all-trans long linear polyenes: the case of the 3,20 di(tert-butyl)-2,2,21,21-tetramethyl-all-trans-3,5,7,9,11,13,15,17,19 docosanonaen (ttbp9) conforming to Kasha's rule. AB - The absorption, fluorescence and fluorescence excitation spectra for 3,20-di(tert butyl)-2,2,21,21-tetramethyl-all-trans-3,5,7,9,11,13,15,17,19-docosanonaen (ttbP9) in dilute solutions of 2-methylbutane were recorded at temperatures over the range 120-280 K. The high photostability of this nonaene allows us to assert that it exhibits a single fluorescence and that this can be unequivocally assigned to emission from its 1(1)B(u) excited state, it being the first excited electronic state. Available photophysical data for this polyene and the wealth of information reported for shorter all-trans polyenes allow us to conclude that if the first excited electronic state for the chromophore possessed 2(1)A(g) symmetry, then the energy of such a state might have been so close to that of the 1(1)B(u) state that: 1) the radiationless internal conversion mechanism would preclude the observation of the emission from the 1(1)B(u) state reported in this work and 2) the 2(1)A(g) state reached through internal conversion would be vibrationally coupled to 1(1)B(u) and would facilitate the detection of the emission from 2(1)A(g), which was not observed in any of the solvents used in this work. The spectroscopic and photochemical implications of these findings for other polyenes are discussed. PMID- 15832402 TI - Resveratrol as an anti-inflammatory and anti-aging agent: mechanisms and clinical implications. AB - Resveratrol is a phytoalexin polyphenolic compound found in various plants, including grapes, berries, and peanuts. Multiple lines of compelling evidence indicate its beneficial effects on neurological, hepatic, and cardiovascular systems. Also one of the most striking biological activities of resveratrol soundly investigated during the late years has been its cancer-chemopreventive potential. In fact, recently it has been demonstrated that this stilbene blocks the multistep process of carcinogenesis at various stages: tumor initiation, promotion, and progression. One of the possible mechanisms for its biological activities involves downregulation of the inflammatory response through inhibition of synthesis and release of pro-inflammatory mediators, modification of eicosanoid synthesis, inhibition of activated immune cells, or inhibiting such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) via its inhibitory effects on nuclear factor (kappa)B (NF-(kappa)B) or the activator protein-1 (AP-1). More recent data provide interesting insights into the effect of this compound on the lifespan of yeast and flies, implicating the potential of resveratrol as an anti-aging agent in treating age-related human diseases. It is worthy to note that the phenolic compound possesses a low bioavailability and rapid clearance from the plasma. As the positive effects of resveratrol on inflammatory response regulation may comprise relevant clinical implications, the purpose of this article is to review its strong anti-inflammatory activity and the plausible mechanisms of these effects. Also, this review is intended to provide the reader an up-date of the bioavailability and pharmacokinetics of resveratrol and its impact on lifespan. PMID- 15832403 TI - Exo-mechanism proximity-accelerated alkylations: investigations of linkers, electrophiles and surface mutations in engineered cyclophilin-cyclosporin systems. AB - Investigations on the scope and utility of exo-mechanism proximity-accelerated reactions in engineered receptor-ligand systems are reported. We synthesized a series of electrophilic cyclosporin (CsA) derivatives by varying electrophiles and linker lengths, prepared a series of nucleophilic cysteine mutations on the surface of cyclophilin A (Cyp), and examined their reactivity and specificity in proximity-accelerated reactions. Acrylamide and epoxide electrophiles afforded useful reactivity and high specificity for alkylation of engineered receptors in Jurkat cell extracts. We found that remote cysteines (>17 A from the ligand) could be alkylated with useful rates under physiological conditions. The results from mutations of the receptor surface suggest that the dominant factors governing the rates of proximity-accelerated reactions are related to the local environment of the reactive group on the protein surface. This study defines several parameters affecting reactivity in exo-mechanism proximity-accelerated reactions and provides guidance for the design of experiments for biological investigations involving proximity-accelerated reactions. PMID- 15832405 TI - No association of the matrix metalloproteinase 1 promoter polymorphism with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in northern China. AB - AIM: To investigate association of the 2G or 1G single nucleotide polymorphism (SNP) in matrix metalloproteinase 1 (MMP1) promoter with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of North China. METHODS: MMP1 promoter SNP was genotyped by polymerase-chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis in 417 cancer patients (234 ESCC and 183 GCA) and 350 healthy controls. RESULTS: The genotype frequencies of the MMP1 promoter SNP in healthy controls were 55.4% (2G/2G), 30% (1G/2G) and 14.6% (1G/1G), respectively. The genotype and allelotype distribution in ESCC and GCA patients was not significantly different from that in healthy controls (all P values were above 0.05). Compared with the 1G/1G genotype, neither the 2G/2G nor in combination with the 1G/2G genotype significantly modified the risk of developing ESCC and GCA, the adjusted odds ratio was 1.28 (95%CI = 0.78-2.09), 1.23 (95%CI = 0.38-2.05) in ESCC and 1.39 (95%CI = 0.80-2.41), 1.34 (95%CI = 0.74-2.40) in GCA, respectively. When stratified by smoking status and family history of upper gastrointestinal cancer, the 2G/2G genotype alone or in combination with the 1G/2G genotype also did not show any significant influence on the risk of ESCC and GCA development. In addition, influence of the MMP1 SNP on lymphatic metastasis in ESCC and GCA was also not observed. CONCLUSION: The 2G or 1G SNP in the MMP1 promoter might not modify the risk of ESCC and GCA development and might not be used as a stratification marker to predict the potential of lymphatic metastasis in these two tumor types. PMID- 15832404 TI - Radical induction theory of ulcerative colitis. AB - To propose a new pathogenesis called Radical Induction to explain the genesis and progression of ulcerative colitis (UC). UC is an inflammatory bowel disease. Colonic inflammation in UC is mediated by a buildup of white blood cells (WBCs) within the colonic mucosal lining; however, to date there is no answer for why WBCs initially enter the colonic mucosa to begin with. A new pathogenesis termed "Radical Induction Theory" is proposed to explain this and states that excess un neutralized hydrogen peroxide, produced within colonic epithelial cells as a result of aberrant cellular metabolism, diffuses through cell membranes to the extracellular space where it is converted to the highly damaging hydroxyl radical resulting in oxidative damage to structures comprising the colonic epithelial barrier. Once damaged, the barrier is unable to exclude highly immunogenic fecal bacterial antigens from invading the normally sterile submucosa. This antigenic exposure provokes an initial immune response of WBC infiltration into the colonic mucosa. Once present in the mucosa, WBCs are stimulated to secrete toxins by direct exposure to fecal bacteria leading to mucosal ulceration and bloody diarrhea characteristic of this disease. PMID- 15832406 TI - Gene expression profile differences in gastric cancer, pericancerous epithelium and normal gastric mucosa by gene chip. AB - AIM: To study the difference of gene expression in gastric cancer (T), pericancerous epithelium (P) and normal tissue of gastric mucosa (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray. METHODS: U133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results. RESULTS: When gastric cancer was compared with normal gastric mucosa, 766 genes were found, with a difference of more than four times in expression levels. Of the 766 genes, 530 were up-regulated (Signal Log Ratio (SLR) >2), and 236 were down regulated (SLR<-2). When pericancerous epithelium was compared with normal gastric mucosa, 64 genes were found, with a difference of more than four times in expression levels. Of the 64 genes, 50 were up-regulated (SLR>2), and 14 were down-regulated (SLR<-2). Compared with normal gastric mucosa, a total of 143 genes with a difference in expression levels (more than four times, either in cancer or in pericancerous epithelium) were found in gastric cancer (T) and pericancerous epithelium (P). Of the 143 genes, 108 were up-regulated (SLR>2), and 35 were down-regulated (SLR<-2). CONCLUSION: To apply a gene chip could find 143 genes associated with the genes of gastric cancer in pericancerous epithelium, although there were no pathological changes in the tissue slices. More interesting, six genes of pericancerous epithelium were up-regulated in comparison with genes of gastric cancer and three genes were down-regulated in comparison with genes of gastric cancer. It is suggested that these genes may be related to the carcinogenesis and development of early gastric cancer. PMID- 15832407 TI - Expression of beta-catenin in hepatocellular carcinoma. AB - AIM: The beta-catenin has been recognized as a critical member of the Wnt signaling pathway and plays an important role in the generation/differentiation of many tissues. Inappropriate activation of this pathway has been implicated in carcinogenesis. The mechanism underlying the development as well as its prognosis of hepatocellular carcinoma (HCC) has remained unclear. The purpose of this study is to analyze the expression of beta-catenin in HCC in relation to histological grades and viral hepatitis backgrounds. METHODS: Thirty-two sections were selected at random from autopsy and surgical cases of HCC. Immunohistologically, the location and positivity of beta-catenin expression in HCC was examined. RESULTS: Normal hepatocytes did not express beta-catenin. In 78% of HCC beta catenin was expressed at the membrane of the cells, with or without cytoplasmic and/or nuclear expression. The tumor cells with well- and moderately differentiated grades expressed frequently at the membrane and cytoplasm compared with poorly-differentiated type. Nuclear expression of beta-catenin was prone to occur in the tumor cells of poorly-differentiated grade. There were 15% of hepatitis C virus (HCV) backgrounds with nuclear expression. In contrast, there was 38% with nuclear expression in hepatitis B virus (HBV) backgrounds. In nonB nonC hepatitis, no case expressed nuclear beta-catenin. CONCLUSION: The beta catenin expression in HCC cells was heterogeneous among types of hepatitis viral infection. Wnt signaling pathway might be deeply involved in less-differentiated HCC and HBV background. PMID- 15832408 TI - Differentiation between malignant and benign nodules in the liver: use of contrast C3-MODE technology. AB - AIM: To investigate the value of contrast-enhanced C3-MODE technology in differentiating malignant nodules of liver from the benign ones. METHODS: Forty six nodules in 36 patients (29 men and 7 women) were studied by contrast-enhanced C3-MODE technology and contrast-enhanced CT in 1 wk before the biopsy or operation. A low MI monitor and a high MI flash imaging were intermittently performed. After the injection of contrast agent, the period from 10 to 30 s and the time later than 100 s were respectively defined as early arterial phase and the late phase. The vascularities of the liver nodules in the two phases were combined for differential diagnosis. Corresponding to the pathological diagnosis, the accuracy, sensitivity and specificity of contrast-enhanced C3-MODE technology were compared to those of contrast-enhanced CT. RESULTS: By C3-MODE technology, 33 of the 46 liver nodules were demonstrated as defected area in the late phase and were diagnosed as malignant tumors. Of them, 28 with hypervascularity in the early arterial phase were assessed as hepatocellular carcinoma, the other five nodules with rim-like enhancement in the early arterial phase were diagnosed as metastatic tumors. Thirteen nodules were shown as iso or hypervascularity in the late phase as well as centripetal filling in the early arterial phase and we made a diagnosis of hemangioma. Corresponding to the pathological results, the sensitivity, specificity and accuracy of contrast-enhanced C3-MODE technology in differentiating malignant and benign nodules in the liver were 97.0%, 92.3% and 95.7%, respectively. With comparison to those of contrast CT (sensitivity, 94.1%; specificity, 91.7%; accuracy, 93.5%), the difference was not significant. CONCLUSION: Contrast-enhanced C3-MODE technology can effectively differentiate malignant liver tumors from the benign nodules. It highly agrees diagnostically with the pathology. We suggest that it provides a new approach for differential diagnosis of liver nodules in addition to contrast-enhanced CT. PMID- 15832409 TI - Hepatic artery infusion of antisense oligodeoxynucleotide and lipiodol mixture transfect liver cancer in rats. AB - AIM: To study the distribution and stability of antisense oligodeoxynucleotide (ASODN) in Walker-256 cells and their distribution in liver, lung and kidney tissues after being infused alone or mixed with lipiodol via hepatic artery in a rat liver tumor model. METHODS: 5'-Isothiocyanate (FITC)-labeled vascular endothelial growth factor (VEGF) ASODN was added into Walker-256 cell culture media. Its distribution in cells was observed by fluorescence microscope at different time points. Walker-256 carcinosarcoma was transplanted into Wistar rat liver to establish a liver cancer model. 5'-FITC-labeled VEGF ASODN mixed with (mixed group, n = 6) or without (TAI group, n = 6) ultra-fluid lipiodol was administrated via hepatic artery. Frozen samples of liver, lung and kidney tissue were taken from rats after 1, 3 and 6 d, respectively. The distribution of ASODN was observed under fluorescent microscope. RESULTS: ASODN could enter cytoplasm within 2 h and nuclei within 6 h. Accumulation of ASODN reached the peak point in nuclei at 12 h, and then disappeared gradually. No fluorescence could be seen in cells at 48 h. In vivo experiment, on d 1 and 3 the fluorescence staining in liver was stronger in mixed group than in TAI group and more fluorescence could be detected in lung and kidney in TAI group than in mixed group. On d 6, no fluorescence could be detected in TAI group, but faint fluorescence could be seen in mixed group. ASODN could be seen in cancer cells and normal hepatic cells. In mixed group, ASODN was mainly distributed in liver tumor tissues. CONCLUSION: ASODN can transfect Walker-256 cells. ASODN mixed with lipiodol infusion via hepatic artery can be used in the treatment of HCC. PMID- 15832410 TI - Determination of optical properties of normal and adenomatous human colon tissues in vitro using integrating sphere techniques. AB - AIM: The purpose of the present study is to compare the optical properties of normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion in vitro at 476.5, 488, 496.5, 514.5 and 532 nm. We believe these differences in optical properties should help differential diagnosis of human colon tissues by using optical methods. METHODS: In vitro optical properties were investigated for four kinds of tissues: normal human colon mucosa/submucosa and muscle layer/chorion, and adenomatous human colon mucosa/submucosa and muscle layer/chorion. Tissue samples were taken from 13 human colons (13 adenomatous, 13 normal). From the normal human colons a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion), and from the adenomatous human bladders a total of 26 tissue samples, with a mean thickness of 0.40 mm, were used (13 from mucosa/submucosa and 13 from muscle layer/chorion). The measurements were performed using a double-integrating-sphere setup and the optical properties were assessed from these measurements using the adding doubling method that was considered reliable. RESULTS: The results of measurement showed that there were significant differences in the absorption coefficients and scattering coefficients between normal and adenomatous human colon mucosa/submucosa at the same wavelength, and there were also significant differences in the two optical parameters between both colon muscle layer/chorion at the same wavelength. And there were large differences in the anisotropy factors between both colon mucosa/submucosa at the same wavelength, there were also large differences in the anisotropy factors between both colon muscle layer/chorion at the same wavelength. There were large differences in the value ranges of the absorption coefficients, scattering coefficients and anisotropy factors between both colon mucosa/submucosa, and there were also large differences in these value ranges between both colon muscle layer/chorion. There are the same orders of magnitude in the absorption coefficients for four kinds of colon tissues. The scattering coefficients of these tissues exceed the absorption coefficients by at least two orders of magnitude. CONCLUSION: There were large differences in the three optical parameters between normal and adenomatous human colon mucosa/submucosa at the same laser wavelength, and there were also large differences in these parameters between both colon muscle layer/chorion at the same laser wavelength. Large differences in optical parameters indicate that there were large differences in compositions and structures between both colon mucosa/submucosa, and between both colon muscle layer/chorion. Optical parameters for four kinds of colon tissues are wavelength dependent, and these differences would be useful and helpful in clinical applications of laser and tumors photodynamic therapy (PDT). PMID- 15832411 TI - Two-dimensional polyacrylamide gel electrophoresis analysis of indomethacin treated human colon cancer cells. AB - AIM: To establish the two-dimensional gel electrophoresis (2-DE) profiles of indomethacin (IN)-treated human colon cancer cell line HCT116, and to provide a new way to study its anti-tumor molecular mechanism through analyzing a variety of protein maps. METHODS: Two-DE profiles of HCT116 were established in IN treated and untreated groups. Total proteins were separated by immobilized pH gradient-based 2-DE. The gels were stained by silver, scanned by ImageScanner, and analyzed with Image Master software. RESULTS: Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated group. The average deviation of spot position was 0.896+/-0.177 mm in IEF direction and 1.106+/-0.289 mm in SDS-PAGE direction respectively. In IN treated group, 1169+/-36 spots were detected and 1061+/-32 spots were matched, the average matching rate was 90.6% in three gels. In untreated group, 1256+/-50 spots were detected and 1168+/-46 spots were matched, the average matching rate was 93.0% in three gels. Forty-five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 protein spots decreased and 9 showed higher expression in IN-treated group, and only two protein spots showed an expression in untreated cells. CONCLUSION: Two-DE profiles of IN-treated and untreated HCT116 cells were established. Apparent 45 different protein spots were detected in IN-treated and untreated HCT116 cells. The analysis on differential protein spots may serve as a new way to study the molecule mechanism of IN treated colon cancer. PMID- 15832412 TI - Effects and possible anti-tumor immunity of electrochemotherapy with bleomycin on human colon cancer xenografts in nude mice. AB - AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establish the experimental basis for clinical application of ECT. METHODS: Forty nude mice, inoculated subcutaneously human colon cancer cell line LoVo for 3 wk, were allocated randomly into four groups: B+E+ (ECT), B+E- (administration of bleomycin alone), B-E+ (administration of electric pulses alone), and B-E- (no treatment). Tumor volumes were measured daily. The animals were killed on the 7th d, the weights of xenografts were measured, and histologies of tumors were evaluated. Cytotoxicity of spleen natural killer (NK) and lymphokine-activated killer (LAK) cells was then assessed by lactic dehydrogenase release assay. RESULTS: The mean tumor volume of group B+E+ was statistically different from the other three groups after the treatment (F = 36.80, P<0.01). There was one case of complete response, seven cases of partial response (PR) in group B+E+, one case of PR in group B+E- and group B-E+ respectively, and no response was observed in group B-E-. The difference of response between group B+E+ and the other three groups was statistically significant (chi2 = 25.67, P<0.01). Histologically, extensive necrosis of tumor cells with considerable vascular damage and inflammatory cells infiltration were observed in group B+E+. There was no statistical difference between the cytotoxicity of NK and LAK cells in the four treatment groups. CONCLUSION: ECT significantly enhances the chemosensitivity and effects of chemotherapy in human colon cancer xenografts in nude mice, and could be a kind of novel treatment modality for human colon cancer. The generation of T-cell dependent, tumor-specific immunity might be involved in the process of ECT. PMID- 15832413 TI - Effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells. AB - AIM: To investigate the effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells in vitro and its anticancer mechanism. METHODS: Human gallbladder carcinoma GBC-SD cells were cultured by cell culture technique. The growth and the invasiveness of GBC-SD cells in vitro were evaluated by the tetrazolium-based colorimetric assay and by the Matrigel experiment and the crossing-river test. Expression of PCNA, Ki-67, MMP2 and TIMP2 proteins of GBC-SD cells was determined by streptavidin-biotin complex method. RESULTS: In vitro norcantharidin inhibited the growth and proliferation of GBC-SD cells in a dose- and time-dependent manner, with the IC50 value of 56.18 microg/mL at 48 h. Norcantharidin began to inhibit the invasion of GBC-SD cells at the concentration of 5 microg/mL, and the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly at 40 microg/mL. After treatment with norcantharidin, the expression of PCNA, Ki-67, and MMP2 was significantly decreased. With the increase in TIMP2 expression, the MMP2 to TIMP2 ratio was decreased significantly (P<0.05). CONCLUSION: Norcantharidin inhibits the proliferation and growth of human gallbladder carcinoma cells in vitro at relatively low concentrations by inhibiting PCNA and Ki-67 expression. Its anti-invasive activity may be the result of decrease in MMP2 to TIMP2 ratio and reduced cell motility. PMID- 15832414 TI - Effects of augmentation of liver regeneration recombinant plasmid on rat hepatic fibrosis. AB - AIM: To investigate the effects of eukaryotic expression of plasmid on augmentation of liver regeneration (ALR) in rat hepatic fibrosis and to explore their mechanisms. METHODS: Ten rats were randomly selected from 50 Wistar rats as normal control group. The rest were administered intraperitoneally with porcine serum twice weekly. After 8 wk, they were randomly divided into: model control group, colchicine group (Col), first ALR group (ALR1), second ALR group (ALR2). Then colchicine ALR recombinant plasmid were used to treat them respectively. At the end of the 4th wk, rats were killed. Serum indicators were detected and histopathological changes were graded. Expression of type I, III, collagen and TIMP-1 were detected by immunohistochemistry and expression of TIMP-1 mRNA was detected by semi-quantified RT-PCR. RESULTS: The histologic examination showed that the degree of the rat hepatic fibrosis in two ALR groups was lower than those in model control group. Compared with model group, ALR significantly reduced the serum levels of ALT, AST, HA, LN, PCIII and IV (P<0.05). Immunohistochemical staining showed that expression of type I, III, collagen and TIMP-1 in two ALR groups was ameliorated dramatically compared with model group (I collagen: 6.94+/-1.42, 5.80+/-1.66 and 10.83+/-3.58 in ALR1, ALR2 and model groups, respectively; III collagen: 7.18+/-1.95, 4.50+/-1.67 and 10.25+/-2.61, respectively; TIMP-1: 0.39+/-0.05, 0.20+/-0.06 and 0.53+/-0.12, respectively, P<0.05 or P<0.01). The expression level of TIMP-1 mRNA in the liver tissues was markedly decreased in two ALR groups compared with model group (TIMP-1 mRNA/beta actin: 0.89+/-0.08, 0.65+/-0.11 and 1.36+/-0.11 in ALR1, ALR2 and model groups respectively, P<0.01). CONCLUSION: ALR recombinant plasmid has beneficial effects on rat hepatic fibrosis by enhancing regeneration of injured liver cells and inhibiting TIMP-1 expressions. PMID- 15832415 TI - Effects of pharmacological serum from normal and liver fibrotic rats on HSCs. AB - AIM: To make drug sera of Salvia miltiorrhiza and Yigankang, both of which are Chinese herbs that activate bleeding and eliminate stasis, in normal rats and those with liver fibrosis, respectively. To investigate and compare the effects of the two different drug sera on the proliferation and activation of hepatic stellate cells (HSCs). METHODS: Some rats were induced with liver fibrosis: 40% carbon tetrachloride (CCl4) subcutaneous injection, twice a week for 9 wk. Salvia miltiorrhiza, Yigankang, colchicines and normal saline were administered into the stomachs of normal rats and those with liver fibrosis. Drug sera were extracted 5 d later. HSCs in vitro were cultivated in different drug sera for 24 h. The rates of proliferation and expression of alpha-smooth muscle actin (alpha-SMA) were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and immunocytochemistry stain, respectively. RESULTS: The drug sera from normal and liver fibrotic rats could be used to cultivate HSCs and to observe the effects of the corresponding components of herbs on HSCs. Salvia miltiorrhiza and Yigankang had better inhibitory effects on HSCs than colchicines (MTT: normal drug serum: Salvia miltiorrhiza 0.42+/-0.08, Yigankang 0.32+/-0.10 vs colchicines 0.45+/-0.12 pathological drug serum: Salvia miltiorrhiza 0.33+/-0.02, Yigankang 0.26+/-0.01 vs colchicines 0.41+/-0.09. P<0.05). The drug sera of Salvia miltiorrhiza, Yigankang from liver fibrotic rats had a stronger inhibitory effect than the same ones from normal rats (MTT: Salvia miltiorrhiza: normal drug serum 0.42+/-0.08 vs pathological drug serum 0.33+/-0.02. Yigankang: normal drug serum 0.32+/-0.10 vs pathological drug serum 0.26+/-0.01. P<0.05). CONCLUSION: Salvia miltiorrhiza and Yigankang could inhibit the expression of alpha-SMA and the proliferation of HSCs. The drug sera from normal and liver fibrotic rats had different effects on HSCs, probably due to different metabolic processes, effective components and different quantities of drug contents in drug sera from rats with different states of liver. PMID- 15832416 TI - Risk-adjustment in hepatobiliary pancreatic surgery. AB - AIM: The present study evaluates the performance of the POSSUM, the American Society of Anesthetists (ASA), APACHE and Childs classification in predicting mortality and morbidity in hepatopancreaticobiliary (HPB) surgery. We describe especially the limitations and advantages of risk in stratifying the patients. METHODS: We investigated 177 randomly chosen patients undergoing elective complex HPB surgery in a single institution with a total of 71 pre-operative and intra operative risk factors. Primary endpoint was in-hospital mortality and morbidity. Ordered logistic regression analysis was used to identify individual predictors of operative morbidity and mortality. RESULTS: The operative mortality in the series was 3.95%. This compared well with the p-POSSUM and APACHE predicted mortality of 4.31% and 4.29% respectively. Post-operative complications amounted to 45% with 24 (13.6%) patients having a major adverse event. On multivariate analysis the pre-operative POSSUM physiological score (OR = 1.18, P = 0.009) was superior in predicting complications compared to the ASA (P = 0.108), APACHE (P = 0.117) or Childs classification (P = 0.136). In addition, serum sodium, creatinine, international normalized ratio (INR), pulse rate, and intra-operative blood loss were independent risk factors. A combination of the POSSUM variables and INR offered the optimal combination of risk factors for risk prognostication in HPB surgery. CONCLUSION: Morbidity for elective HPB surgery can be accurately predicted and applied in everyday surgical practice as an adjunct in the process of informed consent and for effective allocation of resources for intensive and high-dependency care facilities. PMID- 15832417 TI - Risk factors of pancreatic leakage after pancreaticoduodenectomy. AB - AIM: To analyze the risk factors for pancreatic leakage after pancreaticoduodenectomy (PD) and to evaluate whether duct-to-mucosa pancreaticojejunostomy could reduce the risk of pancreatic leakage. METHODS: Sixty-two patients who underwent PD at our hospital between January 2000 and November 2003 were reviewed retrospectively. The primary diseases of the patients included pancreas cancer, ampullary cancer, bile duct cancer, islet cell cancer, duodenal cancer, chronic pancreatitis, pancreatic cystadenoma, and gastric cancer. Standard PD was performed for 25 cases, PD with extended lymphadenectomy for 27 cases, pylorus-preserving PD for 10 cases. A duct-to-mucosa pancreaticojejunostomy was performed for patients with a hard pancreas and a dilated pancreatic duct, and a traditional end-to-end invagination pancreaticojejunostomy for patients with a soft pancreas and a non-dilated duct. Patients were divided into two groups according to the incidence of postoperative pancreaticojejunal anastomotic leakage: 10 cases with leakage and 52 cases without leakage. Seven preoperative and six intraoperative risk factors with the potential to affect the incidence of pancreatic leakage were analyzed with SPSS10.0 software. Logistic regression was then used to determine the effect of multiple factors on pancreatic leakage. RESULTS: Of the 62 patients, 10 (16.13%) were identified as having pancreatic leakage after operation. Other major postoperative complications included delayed gastric emptying (eight patients), abdominal bleeding (four patients), abdominal abscess (three patients) and wound infection (two patients). The overall surgical morbidity was 43.5% (27/62). The hospital mortality in this series was 4.84% (3/62), and the mortality associated with pancreatic fistula was 10% (1/10). Sixteen cases underwent duct-to-mucosa pancreaticojejunostomy and 1 case (1/16, 6.25%) developed postoperative pancreatic leakage, 46 cases underwent invagination pancreaticojejunostomy and 9 cases (9/46, 19.6%) developed postoperative pancreatic leakage. General risk factors including patient age, gender, history of jaundice, preoperative nutrition, pathological diagnosis and the length of postoperative stay were similar in the two groups. There was no statistical difference in the incidence of pancreatic leakage between the patients who received the prophylactic use of octreotide after surgery and the patients who did not undergo somatostatin therapy. Moreover, multivariate logistic regression analysis showed that none of the above factors seemed to be associated with pancreatic fistula. Two intraoperative risk factors, pancreatic duct size and texture of the remnant pancreas, were found to be significantly associated with pancreatic leakage. The incidence of pancreatic leakage was 4.88% in patients with a pancreatic duct size greater than or equal to 3 mm and was 38.1% in those with ducts smaller than 3 mm (P = 0.002). The pancreatic leakage rate was 2.94% in patients with a hard pancreas and was 32.1% in those with a soft pancreas (P = 0.004). Operative time, blood loss and type of resection were similar in the two patient groups. The incidence of pancreatic leakage was 6.25% (1/16) in patients with duct-to-mucosa anastomosis, and was 19.6% (9/46) in those with traditional invagination anastomosis. Although the difference of pancreatic leakage between the two groups was obvious, no statistical significance was found. This may be due to the small number of patients with duct-to-mucosa anastomosis. By further analyzing with multivariate logistic regression, both pancreatic duct size and texture of the remnant pancreas were demonstrated to be independent risk factors (P = 0.007 and 0.017, OR = 11.87 and 15.45). Although anastomotic technique was not a significant factor, pancreatic leakage rate was much less in cases that underwent duct-to-mucosa pancreaticojejunostomy. CONCLUSION: Pancreatic duct size and texture of the remnant pancreas are risk factors influencing pancreatic leakage after PD. Duct-to-mucosa pancreaticojejunostomy, as a safe and useful anastomotic technique, can reduce pancreatic leakage rate after PD. PMID- 15832418 TI - Characteristics and therapeutic efficacy of sulfasalazine in patients with mildly and moderately active ulcerative colitis. AB - AIM: To investigate the characteristics and short-term efficacy of sulfasalazine (SASP) in patients with mildly and moderately active ulcerative colitis (UC). METHODS: Two hundred and twenty-eight patients with mildly and moderately active UC were recruited, 106 patients in 1993-1995, and 122 patients in 2000-2002, they were assigned as the 1990s group (n = 106) and the 2000s group (n = 122), prospectively. The general characteristics, clinical manifestations, colonoscopic and histological data were compared between the two groups. The short-term efficacy and safety of SASP 3 g per d were evaluated. RESULTS: Between 2000s and 1990s groups, the gender ratio of men to women was 1:1.18 and 1:1.04, 57.4% and 50.9% of the patients were between 30 and 49 years old. The gender ratio and age of UC patients were not significantly different. The total course of 50.0% and 37.1% of UC patients was less than 1 year (P<0.05), 10.6% and 31.2% of the cases had a duration of more than 5 years (P<0.05) in 2000s and 1990s groups, respectively. The most common clinical type was first episode in 2000s group and chronic relapse in 1990s group. The patients showed a higher frequency of abdominal pain and tenderness in 1990s group than in 2000s group. Erosions were found in 84.4% and 67.9% of patients in 2000s and 1990s groups (P<0.05). Rough and granular mucosa (67.9% vs 43.4%, P<0.05) and polyps (47.2% vs 32.8%, P<0.05) were identified in 1990s group more than in 2000s group. There were no significant differences in clinical, colonoscopic and histological classifications. After SASP (1 g thrice per d) treatment for 6 wk, the clinical, colonoscopic and histological remission rates were 71.8%, 21.8% and 16.4%, respectively. In 79 patients with clinical remission, 58.2% and 67.1% remained grade 1 in colonoscopic and histological findings, respectively. The overall effects in first episode type (complete remission in 10, 18.9%, partial remission in 28, 52.8%, and improvement in 9, 17.0%) were better than in chronic relapse type (complete remission in 3, 7.5%; partial remission in 16, 40.0%; and improvement in 15, 37.5%) and chronic persistent type (complete remission in 1, 5.9%; partial remission in 6, 35.3%; and improvement in 6, 35.3%) respectively (P<0.05). In 110 patients treated with SASP, 18 patients (16.4%) had adverse reactions. Except for two cases of urticaria and one case of WBC decrease, none of the patients had to stop the treatment because of severe adverse reactions. CONCLUSION: Patients with mildly and moderately active UC in 2000s group had a shorter disease course, milder clinical manifestations, more first episode type and higher frequency of acute mucosal lesions in colonoscopy than in 1990s group. The patients in 1990s group had higher proportion of chronic relapse type and chronic mucosal change in colonoscopy than in 2000s group. The short-term efficacy of SASP could be mainly remission of clinical manifestations. But more than half of the patients still had light inflammation in colonoscopy and histology. The overall effects of SASP in first episode type were better than those in other types. SASP was a safe and effective drug to treat mildly and moderately active UC. PMID- 15832419 TI - Extended radical operation of pancreatic head cancer: appraisal of its clinical significance. AB - AIM: To evaluate the significance of extended radical operation and its indications. METHODS: Between January 1995 and December 1998, 56 inpatients with pancreatic head cancer received operation. Among them 35 patients (group 1) experienced the Whipple operation, and 21 patients (group 2) received the extended radical operation. The 1-, 2-, 3-year cumulative survival rates were used to evaluate the efficacy of the two operative procedures. Clinical stage (CS) was assessed retrospectively with the help of CT. The indications for extended radical operation were discussed. RESULTS: There was no difference in hospital mortality and morbidity rates. Whereas the 1-, 2-, 3-year cumulative survival rates were 84.8%, 62.8%, 39.9% in the extended radical operation group, and were 70.8%, 47.6%, 17.2% in the Whipple operation group, there was a significant difference between the two groups (P<0.001, P<0.001, P<0.001, respectively). Most of the deaths within 3 years after operation were due to recurrence in the two groups. However, the 1-, 2-, 3-year cumulative rates of death due to local recurrence were decreased from 37.4% in patients that received the Whipple procedure to 23.8% in those who received by extended radical operation. Patients who survived for more than 3 years were only noted in those with CS1 in the Whipple procedure group and were founded in cases with CS1, CS2 and part of CS3 in the extended radical operation group. CONCLUSION: The extended radical operation appears to benefit patients with pancreatic head carcinoma which was indicated in CS1, CS2 and part of CS3 without severe invasion. PMID- 15832420 TI - Intrabiliary rupture: an algorithm in the treatment of controversial complication of hepatic hydatidosis. AB - AIM: Intrabiliary rupture (IBR) is a common and serious complication of hepatic hydatid cyst. The incidence varies from 1% to 25%. The treatment of IBR is still controversial. We aimed to design an algorithm for the treatment of hepatic hydatidosis with IBR by reviewing our cases. METHODS: Eight cases of IBR were analyzed retrospectively. Patients were evaluated according to age, sex, clinical findings, cyst number and stage, abdominal ultrasonography and CT-scan, surgical methods, complications, results and coincidental diseases. RESULTS: Female/male ratio was 1/7. Mean age was 52.12+/-18.26 years (range 24-69 years). Right upper quadrant pain, flatulence, palpable hepatic mass were symptoms common in all patients. Cholestatic jaundice was found in four cases. In all patients, cyst evacuation and omentoplasty were performed, followed by either choledochoduodenostomy, T-tube drainage, intracavitary suturing of the orifice, two cases in each. Whereas in two patients diagnosed post-operatively percutaneous drainage of biliary collection or ERCP and sphincteroplasty were added. Morbidity and hospital stay were higher in these cases. CONCLUSION: When the diagnosis of IBR can be done pre- or intra-operatively, morbidity decreases. If a biliary fistula is seen post-operatively, endoscopic procedures such as ERCP, sphincteroplasty or nasobiliary drainage can be applied. PMID- 15832421 TI - Efficacy of omeprazole and amoxicillin with either clarithromycin or metronidazole on eradication of Helicobacter pylori in Chinese peptic ulcer patients. AB - AIM: One-week triple therapy with proton pump inhibitors, clarithromycin and amoxicillin has recently been proposed as the first-line treatment for Helicobacter pylori (H. pylori) infection; however, data regarding the effects of this regimen in China are scarce. The aim of this prospective and randomized study was to compare the efficacy of clarithromycin and metronidazole when they were combined with omeprazole and amoxicillin on eradication of H. pylori and ulcer healing in Chinese peptic ulcer patients. METHODS: A total of 103 subjects with H. pylori-positive peptic ulcer were randomly divided into two groups, and accepted triple therapy with omeprazole 20 mg, amoxicillin 1000 mg and either clarithromycin 500 mg (OAC group, n = 58) or metronidazole 400 mg (OAM group, n = 45). All drugs were given twice daily for 7 d. Patients with active peptic ulcer were treated with omeprazole 20 mg daily for 2-4 wk after anti-H. pylori therapy. Six to eight weeks after omeprazole therapy, all patients underwent endoscopies and four biopsies (two from the antrum and two others from the corpus of stomach) were taken for rapid urease test and histological analysis (with modified Giemsa staining) to examine H. pylori. Successful eradication was defined as negative results from both examination methods. RESULTS: One hundred patients completed the entire course of therapy and returned for follow-up. The eradication rate of H. pylori for the per-protocol analysis was 89.3% (50/56) in OAC group and 84.1% (37/44) in OAM group. Based on the intention-to-treat analysis, the eradication rate of H. pylori was 86.2% (50/58) in OAC group and 82.2% (37/45) in OAM group. There were no significant differences in eradication rates between the two groups on either analysis. The active ulcer-healing rate was 96.7% (29/30) in OAC group and 100% (21/21) in OAM group (per-protocol analysis, P>0.05). Six patients in OAC group (10.3%) and five in OAM group (11.1%) reported adverse events (P>0.05). CONCLUSION: One-week triple therapy with omeprazole and amoxicillin in combination with either clarithromycin or metronidazole is effective for the eradication of H. pylori. The therapeutic regimen comprising metronidazole with low cost, good compliance and mild adverse events may offer a good choice for the treatment of peptic ulcers associated with H. pylori infection in China. PMID- 15832422 TI - Expression of cellular FLICE-inhibitory protein and its association with p53 mutation in colon cancer. AB - AIM: To investigate the expression of cellular FLICE (Fas associated death domain like IL-1beta-converting enzyme)-inhibitory protein (c-FLIP) and its association with p53 mutation in colon cancer. METHODS: Immunohistochemical staining of c FLIP and mutant p53 by using specific antibodies was performed by the standard streptavidin-peroxidase technique for 45 colon cancer tissue samples with matched normal tissues. Semi-quantitative reverse transcriptional (RT)-PCR was used to measure c-FLIP mRNA levels. t-test statistical method was used in data analyses. RESULTS: c-FLIP mRNA was expressed in all colon cancer tissues and its level (0.63+/-0.12) was significantly higher than that in normal tissues (0.38+/-0.10, P<0.01). Immuno-histochemically, c-FLIP protein was also expressed in all colon cancers (45/45) and 71.1% (32/45) showed an intense immunostaining, in contrast, 93.3% (42/45) of normal colonic mucosa showed positive staining and none of them immunostained intensely. The quantity of c-FLIP protein was significantly higher in cancer tissues than in normal mucosa (7.04+/-1.20 vs 5.21+/-0.86, P<0.01). Positive staining of mutant p53 protein was found in 60% (27/45) colon cancers. c FLIP mRNA level was decreased in p53 positive group compared with p53 negative cancer tissues (0.59+/-0.13 vs 0.69+/-0.14, P<0.01), but c-FLIP protein had a significantly higher level in p53 positive cancer tissues than in negative ones (7.57+/-1.30 vs 6.25+/-1.27, P<0.01). CONCLUSION: c-FLIP is specially overexpressed in colon cancers and it might contribute to carcinogenesis of normal colonic mucosa. p53 may exert transcriptional upregulation effects on c FLIP gene and more potent effects on promoting the degradation of c-FLIP protein. PMID- 15832423 TI - Correlation between CD4, CD8 cell infiltration in gastric mucosa, Helicobacter pylori infection and symptoms in patients with chronic gastritis. AB - AIM: To evaluate the correlation between CD4, CD8 cell infiltration in gastric mucosa, Helicobacter pylori (H pylori) infection and symptoms or the assemblage of symptoms in cases with chronic gastritis. METHODS: Biopsy samples at the gastric antrum were obtained from 62 patients with chronic gastritis. CD4 and CD8 cell infiltration was evaluated by immunohistochemical assays on frozen sections of the biopsy samples. Fifteen symptoms referring to digestion-related activity and non-digestion related activity were observed. The correlation between lymphocyte infiltration and each symptom or symptom assemblage was analyzed by logistic regression and K-mean cluster methods. RESULTS: CD4 cell infiltrations in gastric mucosa were much more in patients with H. pylori infection, while CD8 cell infiltrations were similar in patients with or without H. pylori infection. Logistic regression analysis showed that the symptoms including heavy feeling in head or body (t = 2.563), and thirst (t = 2.478) were significantly related with CD4 cell infiltration in gastric mucosa (P<0.05), and cool limbs with aversion to cold were related with CD8 cell infiltration (t = 2.872, P<0.05). Further analysis showed that non-digestive related symptom assemblage could increase the predicted percentage of CD4 and CD8 cell infiltration in gastric mucosa, including lower CD4 infiltration by 12.5%, higher CD8 infiltration by 33.3%, and also non-H. pylori infection by 23.6%. K-means cluster analysis of all symptoms and CD4 and CD8 cell infiltration in gastric mucosa showed a similar tendency to increase the predicted percentage of CD4, CD8 cell infiltration and H. pylori infection. CONCLUSION: Based on correlation between the gastric mucosa lymphocyte infiltration, H. pylori infection and clinical symptoms, symptoms or symptomatic assemblages play an important role in making further classification of chronic gastritis, which might help find a more specific therapy for chronic gastritis. PMID- 15832424 TI - Combined effects of Cantide and chemotherapeutic drugs on inhibition of tumor cells' growth in vitro and in vivo. AB - AIM: To investigate the combination effect of hTERT antisense oligonucleotide "Cantide" and three chemotherapeutic drugs (cisplatin, 5-fluorouracil (5-FU) and adriamycin (ADM)) on inhibiting the proliferation of HepG2, BGC and A549 cell lines in vitro, and to investigate the efficacy of Cantide used in combination with cisplatin (DDP) in vivo. METHODS: Cantide was transfected into these tumor cells by Lipofectin, and cell growth activity was calculated by microcytotoxicity assay. In vivo study, cells of HepG2 were implanted in Balb/c nude mice for 4 d. Then Cantide, DDP and Cantide+DDP were given intraperitoneally for 24 d respectively. The body weights of the tumor-bearing animals and their tumor mass were measured later to assess the effect of combination therapy in the nude mice. To evaluate the interaction of Cantide and these chemotherapeutic drugs, SAS software and Jin Zhengjun method were used. RESULTS: Combination treatments with 0.1 micromol/L Cantide reduced the IC50 of DDP, 5-FU and ADM from 1.07, 4.15 and 0.29 microg/mL to 0.25, 1.52 and 0.12 microg/mL respectively. The inhibition ability of DDP, 5-FU and ADM respectively in combination with Cantide in these tumor cells was higher than that of these drugs alone (P<0.0001). And synergism (Q > or = 1.15) was observed at the lower concentration of DDP (< or = 1 microg/mL), 5-FU (< or = 10 microg/mL) and ADM (< or = 0.1 microg/mL) with combination of Cantide. In vivo, combination treatment with Cantide and DDP produced the greater growth inhibition of human liver carcinoma cells HepG2 in nude mice (0.65+/-0.19 g tumor) compared with that when only one of these drugs was used (Cantide group: 1.05+/-0.16 g tumor, P = 0.0009<0.001; DDP group: 1.13+/ 0.09 g tumor, P = 0.0001<0.001). CONCLUSION: These findings indicate that Cantide may enhance therapeutic effectiveness of chemotherapeutic drugs over a wide range of tumor cells in vitro, and the combination use of Cantide and DDP can produce much higher inhibition rates, as compared with when either of these drugs was used only in vivo. PMID- 15832425 TI - Effective siRNA targets screening for human telomerase reverse transcriptase. AB - AIM: To study the inhibitory effects of siRNAs targeting different hTERT sequences and to screen the effective siRNA sequence. METHODS: Five double stranded siRNAs targeting coding and non-coding regions of hTERT gene were designed and synthesized by T7 transcription system in vitro. siRNA4 sequence was screened by full length gene targeting technique and the rest of the siRNA sequences were selected randomly. After being purified by ethanol precipitation, the siRNAs were transfected to the human hepatocellular carcinoma cell (HepG2) by Lipofectamine 2000. At 48-72 h after siRNAs transfection, MTT assay, RT-PCR and Western-blot were applied to evaluate the effects of siRNAs on cell growth, mRNA and protein expression level of hTERT gene, respectively. RESULTS: Compared to the control cells, the cells treated with the five double-stranded siRNAs exhibited different degrees of inhibition of cell proliferation in a dose dependent manner. siRNA2 and siRNA4, exhibited obvious effects of inhibiting hTERT mRNA and protein expression in HepG2 cells. CONCLUSION: siRNAs targeting different hTERT sequences have significantly various inhibitory effects on hTERT gene expression. The siRNA sequence screened by full length gene targeting technique has comparable inhibitory effect with the rest siRNA sequences screened by random selection, suggesting that siRNAs and antisense oligonucleic acids may have the same effective target sites. Compared with chemical synthesis method, synthesizing double-stranded siRNA by T7 transcription system in vitro is a rapid, simple, and inexpensive method suitable for screening high-effect siRNA targeting site for specific gene. PMID- 15832426 TI - Effects of dendritic cells from cord blood CD34+ cells on human hepatocarcinoma cell line BEL-7402 in vitro and in SCID mice. AB - AIM: To develop a cancer vaccine of dendritic cells derived from human cord blood CD34+ cells and to investigate its cytotoxicity on human hepatocarcinoma cells in vitro and in severe combined immunodeficiency (SCID) mice. METHODS: Lymphocytes from cord blood or peripheral blood were primed by DCs, which were derived from cord blood and pulsed with whole tumor cell lysates. Nonradiative neutral red uptake assay was adopted to detect the cytotoxicity of primed lymphocytes on human hepatocarcinoma cell line BEL-7402 in vitro. The anti-tumor effect of primed lymphocytes in vivo was detected in SCID mice, including therapeutic effect and vaccination effect. RESULTS: The cytotoxicity of DC vaccine primed lymphocytes from cord blood or peripheral blood on human hepatocarcinoma cell line BEL-7402 was significantly higher than that of unprimed lymphocytes in vitro (44.09% vs 14.69%, 47.92% vs 19.44%, P<0.01). There was no significant difference between the cytotoxicity of primed lymphocytes from cord blood and peripheral blood (P>0.05). The tumor growth rate and tumor size were smaller in SCID mice treated or vaccinated with primed lymphocytes than those with unprimed lymphocytes. SCID mice vaccinated with primed lymphocytes had a lower tumor incidence (80% vs 100%, P<0.05) and delayed tumor latent period compared with mice vaccinated with unprimed lymphocytes (11 d vs 7 d, P<0.01). CONCLUSION: Vaccine of cord blood derived-DCs has an inhibitory activity on growth of human hepatocarcinoma cells in vitro and in SCID mice. The results also implicate the potential role of cord blood derived-DC vaccine in clinical tumor immunotherapy. PMID- 15832427 TI - Profiling of differentially expressed chemotactic-related genes in MCP-1 treated macrophage cell line using human cDNA arrays. AB - AIM: To study the global gene expression of chemotactic genes in macrophage line U937 treated with human monocyte chemoattractant protein-1 (MCP-1) through the use of ExpreeChipHO2 cDNA array. METHODS: Total RNA was extracted from MCP-1 treated macrophage line U937 and normal U937 cells, reversely transcribed to cDNA, and then screened in parallel with HO2 human cDNA array chip. The scanned result was additionally validated using RT-PCR. RESULTS: The result of cDNA array showed that one chemotactic-related gene was up-regulated more than two-fold (RANTES) and seven chemotactic-related genes were down-regulated more than two fold (CCR1, CCR5, ccl16, GRObeta, GROgamma, IL-8 and granulocyte chemotactic protein 2) in MCP-1 treated U937 cells at mRNA level. RT-PCR analysis of four of these differentially expressed genes gave results consistent with cDNA array findings. CONCLUSION: MCP-1 could influence some chemokine and receptor expressions in macrophages in vitro. MCP-1 mainly down-regulates the expression of chemotactic genes influencing neutrophilic granulocyte expression (GRObeta, GROgamma, IL-8 and granulocyte chemotactic protein 2), and the mRNA level of CCR5, which plays a critical role in many disorders and illnesses. PMID- 15832428 TI - A randomized controlled trial of laparoscopic versus open cholecystectomy in patients with cirrhotic portal hypertension. AB - AIM: To evaluate the characters, risks and benefits of laparoscopic cholecystectomy (LC) in cirrhotic portal hypertension (CPH) patients. METHODS: Altogether 80 patients with symptomatic gallbladder disease and CPH, including 41 Child class A, 32 Child class B and 7 Child class C, were randomly divided into open cholecystectomy (OC) group (38 patients) and LC group (42 patients). The cohorts were well-matched for number, age, sex, Child classification and types of disease. Data of the two groups were collected and analyzed. RESULTS: In LC group, LC was successfully performed in 36 cases, and 2 patients were converted to OC for difficulty in managing bleeding under laparoscope and dense adhesion of Calot's triangle. The rate of conversion was 5.3%. The surgical duration was 62.6+/-15.2 min. The operative blood loss was 75.5+/-15.5 mL. The time to resume diet was 18.3+/-6.5 h. Seven postoperative complications occurred in five patients (13.2%). All patients were dismissed after an average of 4.6+/-2.4 d. In OC group, the operation time was 60.5+/-17.5 min. The operative blood loss was 112.5+/-23.5 mL. The time to resume diet was 44.2+/-10.5 h. Fifteen postoperative complications occurred in 12 patients (30.0%). All patients were dismissed after an average of 7.5+/-3.5 d. There was no significant difference in operation time between OC and LC group. But LC offered several advantages over OC, including fewer blood loss and lower postoperative complication rate, shorter time to resume diet and shorter length of hospitalization in patients with CPH. CONCLUSION: Though LC for patients with CPH is difficult, it is feasible, relatively safe, and superior to OC. It is important to know the technical characters of the operation, and pay more attention to the meticulous perioperative managements. PMID- 15832429 TI - Effect of anti-tuberculosis therapy on liver function of pulmonary tuberculosis patients infected with hepatitis B virus. AB - AIM: To observe the effect of anti-tuberculosis therapy on liver function of pulmonary tuberculosis patients with hepatitis B virus (HBV) infection, and to compare the differences of liver function by two treatments of anti-tuberculosis. METHODS: Forty-seven TB patients with HBV infection and 170 TB patients without HBV infection were divided into HPBE(S) and HLAMKO treatment groups. Liver function tests before and after the treatments were performed once in 2 wk or monthly, and their clinical manifestations were recorded. RESULTS: The rate of hepatotoxicity occurred in 26 (59%) TB patients with HBV during anti-TB treatment, higher than that in 40 (24%) TB patients without HBV. Hepatotoxicity occurred in 66 out of 217 patients, and the incidence of liver dysfunction was 46.1% in HPBE(S) group, significantly higher than that in HLAMKO group (12.7%) (P<0.01). CONCLUSION: TB patients with HBV should choose HLAMKO treatment because of fewer hepatotoxicity. PMID- 15832430 TI - Evaluation of CMU-1 preservation solutions using an isolated perfused rat liver model. AB - AIM: CMU-1 is a new preservation solution with a low potassium concentration as well as low viscosity that is highly effective in reducing preservation injury. The purpose of this experiment is to compare the protective effect of CMU-1 solution with that of UW during cold preservation and normothermic reperfusion. METHODS: Wistar rats were divided into two groups according to different preservation solution: CMU-1 group and UW group. After 6, 12 and 24 h cold storage of rat liver in different preservation solutions, the isolated perfused rat liver model was applied to reperfuse the liver for 120 min normothermically (37 degrees C) with Krebs-Henseleit solution, meanwhile the pH value of the preservation solution was measured. The perfusate was sampled for the evaluation of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). At the end of the reperfusion, all of the bile product was collected, energy metabolic substrate and histological examination were performed. RESULTS: After preserving for 6 h, pH value of both groups did not change; after 12 h, both decreased but with no significant difference. After 24 h, pH value in UW solution group significantly decreased. The total adenine nucleotides level and AEC in liver tissue decreased with preservation time, but they were higher in CMU-1 group. And the amount of bile product after perfusion for 120 min in CMU-1 group was much more than that in UW group. However, there were no significant differences in ALT and LDH levels between two groups. Histology showed no difference. CONCLUSION: The preservation effect of CMU-1 solution is similar with that of UW solution. However, CMU-1 solution shows some advantages over UW solution in energy metabolism, preventing intracellular acidosis and bile product. PMID- 15832432 TI - Training of a minimally invasive bariatric surgeon: are laparoscopic fellowships the answer? AB - BACKGROUND: Fellowships in advanced laparoscopy with emphasis in laparoscopic gastric bypass (LGBP) are available for obtaining experience in performing LGBP. The following is the first report in the literature prospectively documenting a single surgeon's experience with LGBP outcomes following completion of an advanced laparoscopic surgical fellowship. METHODS: Outcomes measured prospectively included length of stay, length of operation, complications, reduction in obesity-related co-morbidities, and percentage excess weight loss. Outcomes were analyzed by quartile to see if there was difference over time. Complications were also compared to outcomes in the literature. RESULTS: 175 patients (147 female, 28 male) underwent LGBP. The mean BMI was 49.2. Mean operative time was 123 minutes, and mean length of stay was 2.2 days. The percentage excess weight loss at 1 year was 73% (n = 79). One patient developed an internal hernia (0.6%) and 1 patient developed an anastomotic leak (0.6%). Post operative transfusion rate was 4.6%. There were no deep venous thromboses or pulmonary emboli detected. There were no conversions to open, and there was no mortality. Upon quartile analysis, there was no difference in complication rates. Complication rates were comparable to published outcomes in the literature. CONCLUSION: Fellowships in advanced laparoscopy with emphasis on LGBP provide the optimal training environment for acquisition of skills necessary to safely and effectively perform LGBP. With fellowship training, complication rates were comparable to published outcomes in the literature without a period of higher complications (the learning curve). PMID- 15832431 TI - Systemic chemo-immunotherapy for advanced-stage hepatocellular carcinoma. AB - AIM: To evaluate the therapeutic efficacy of systemic chemo-immunotherapy for advanced hepatocellular carcinoma (HCC). METHODS: Twenty-six patients with advanced HCC were treated by using systemic chemo-immunotherapy (PIAF regimen), which consisted of cisplatin (20 mg/m2) intravenously daily for 4 consecutive day, doxorubicin (40 mg/m2) intravenously on day 1, 5-fluorouracil (400 mg/m2) intravenously daily for 4 consecutive day, and human recombinant alpha-interferon 2a (5 MU/m2) subcutaneous injection daily for 4 consecutive day. The treatment was repeated every 3 wk, with a maximum of six cycles. RESULTS: A total of 90 cycles of PIAF treatment were administered, with a mean number of 3.9 cycles per patient. Eight patients received six cycles of treatment (group A), and the remaining 18 were subjected to two to five cycles (group B). There were 0 complete responses, 4 partial responses, 9 static diseases and 13 progressive diseases, with a disease control rate of 50% (13/26). The 1-year survival rate was 24.3%, with a median survival time of 6.0 mo. Group A had a remarkably better survival as compared with group B, the 1- and 2-year survival rates were 62.5% vs 6.1% and 32.3% vs 0%, and a median survival time was 12.5 mo vs 5.0 mo (P = 0.001). CONCLUSION: Systemic chemo-immunotherapy using PIAF regimen represented an effective treatment and could improve the survival rate and prolong the survival time in selected patients with advanced HCC. PMID- 15832433 TI - Screening for prostate cancer. The case against. PMID- 15832434 TI - Crafting interventional procedures guidance. PMID- 15832435 TI - Extremely high body mass index is not a contraindiction to laparoscopic gastric bypass? PMID- 15832436 TI - Sesamoid arthrodesis of the thumb: a technique using a Mitek anchor and wire suture. PMID- 15832437 TI - Revision arthroplasty of a loose acetabular component in the presence of a solid monobloc femoral component: a novel technique. PMID- 15832438 TI - Use of aortic restrictor in intramedullary nailing. PMID- 15832439 TI - Maintaining arthroscopy irrigation pressure without a mechanical pump. PMID- 15832440 TI - Circumferential periosteal block in the reduction of distal radius fractures. PMID- 15832441 TI - Use of Rhys-Davies exsanguinator in positioning for retrograde femoral nailing. PMID- 15832442 TI - How not to be stumped by the traction table. PMID- 15832443 TI - Cytochrome P450: nature's most versatile biological catalyst. AB - The author describes studies that led to the resolution and reconstitution of the cytochrome P450 enzyme system in microsomal membranes. The review indicates how purification and characterization of the cytochromes led to rigorous evidence for multiple isoforms of the oxygenases with distinct chemical and physical properties and different but somewhat overlapping substrate specificities. Present knowledge of the individual steps in the P450 and reductase reaction cycles is summarized, including evidence for the generation of multiple functional oxidants that may contribute to the exceptional diversity of the reactions catalyzed. PMID- 15832444 TI - Formation and toxicity of anesthetic degradation products. AB - Toxic degradation products are formed from a range of old and modern anesthetic agents. The common element in the formation of degradation products is the reaction of the anesthetic agent with the bases in the carbon dioxide absorbents in the anesthesia circuit. This reaction results in the conversion of trichloroethylene to dichloroacetylene, halothane to 2-bromo-2-chloro-1,1 difluoroethylene, sevoflurane to 2-(fluoromethoxy)-1,1,3,3,3-pentafluoro-1 propene (Compound A), and desflurane, isoflurane, and enflurane to carbon monoxide. Dichloroacetylene, 2-bromo-2-chloro-1,1-difluoroethylene, and Compound A form glutathione S-conjugates that undergo hydrolysis to cysteine S-conjugates and bioactivation of the cysteine S-conjugates by renal cysteine conjugate beta lyase to give nephrotoxic metabolites. The elucidation of the mechanisms of formation and bioactivation of degradation products has allowed for the safe use of anesthetics that may undergo degradation in the anesthesia circuit. PMID- 15832445 TI - Non-Michaelis-Menten kinetics in cytochrome P450-catalyzed reactions. AB - The cytochrome P450 monooxygenases (CYPs) are the dominant enzyme system responsible for xenobiotic detoxification and drug metabolism. Several CYP isoforms exhibit non-Michaelis-Menten, or "atypical," steady state kinetic patterns. The allosteric kinetics confound prediction of drug metabolism and drug drug interactions, and they challenge the theoretical paradigms of allosterism. Both homotropic and heterotropic ligand effects are now widely documented. It is becoming apparent that multiple ligands can simultaneously bind within the active sites of individual CYPs, and the kinetic parameters change with ligand occupancy. In fact, the functional effect of any specific ligand as an activator or inhibitor can be substrate dependent. Divergent approaches, including kinetic modeling and X-ray crystallography, are providing new information about how multiple ligand binding yields complex CYP kinetics. PMID- 15832446 TI - Postpartum post-dural puncture headache. PMID- 15832447 TI - Postpartum post-dural puncture headache. PMID- 15832448 TI - Aspiration and the laryngeal mask airway. PMID- 15832449 TI - Aspiration and the laryngeal mask airway. PMID- 15832450 TI - C-reactive protein levels and outcomes after statin therapy. PMID- 15832451 TI - C-reactive protein levels and outcomes after statin therapy. PMID- 15832452 TI - Molecular prediction of recurrence of breast cancer. PMID- 15832453 TI - Molecular prediction of recurrence of breast cancer. PMID- 15832454 TI - Molecular prediction of recurrence of breast cancer. PMID- 15832455 TI - Prophylaxis against rabies. PMID- 15832456 TI - Prophylaxis against rabies. PMID- 15832457 TI - Recombinant hepatitis B vaccine and the risk of multiple sclerosis: a prospective study. PMID- 15832458 TI - Cosmetic neurology: the controversy over enhancing movement, mentation, and mood. PMID- 15832459 TI - [The association between low birthweight and asthma: a systematic literature review]. AB - OBJECTIVE: To determine if there is an association between low birthweight (< 2,500 g) and asthma. RESULTS: The key words "asthma," "children," "birth weight," and "risk factors" were used to identify analytical epidemiological studies on humans that were published in English, Portuguese, or Spanish between 1990 and 2001 and that were indexed in either of two bibliographic databases: MEDLINE (PubMed) and Latin American and Caribbean Health Sciences (Literatura Latino Americana e do Caribe em Ciencias da Saude (LILACS)). We identified 41 articles dealing with the association between birthweight and asthma: 21 were longitudinal studies, 16 were cross-sectional studies, and 4 were case-control studies. Low birthweight was identified as a risk factor for asthma in 26 of the articles. Two studies found an inverse association between birthweight and risk for asthma, and 15 did not find any association. CONCLUSIONS: The association between low birthweight and asthma remains controversial, mainly due to the extensive network of factors that contribute to the development of asthma. The association between low birthweight and asthma occurs mainly among children up to 5 years of age, and it decreases as age increases. Pulmonary function is an important factor that needs to be considered. Low birthweight children may have decreased pulmonary function and consequently develop asthma. On the other hand, these children may have asthma as a consequence of low birthweight, without pulmonary function being a factor. In the six studies comparing low birthweight children (< 2,500 g) with very low birthweight children (< 1,500 g), very low birthweight was associated with a greater risk for developing asthma. Finally, the contribution of intrauterine factors (such as nutrition) in the development of asthma is receiving increasing attention in the scientific literature. Controlling for possible confounding factors is essential to clarifying the relationship between birthweight and asthma. PMID- 15832460 TI - Pharmaceutical application of thermoresponsive materials. PMID- 15832461 TI - Swelling and paracetamol release studies of poly(acrylamide-co-itaconic acid) hydrogels. PMID- 15832462 TI - A green health education approach to health promoting schools. AB - Since the Education Commission of the Hong Kong Special Administrative Region had proposed the educational reform in 2000, there is a new direction for school curriculum development. Lok Wah Catholic Primary School was one of the schools taking the health promoting school approach for educational reform. This paper aims to share information on how the School implemented the "Green Health Education Approach" to create a healthy learning environment for students. PMID- 15832463 TI - Impact of empiric antibiotics and chest radiograph on delays in the diagnosis of tuberculosis. AB - SETTING: Maryland Department of Health and Mental Hygiene, Division of Tuberculosis (TB) Control. OBJECTIVES: To assess the implications of antibiotic treatment of presumed community-acquired pneumonia (CAP) on delays in the diagnosis of TB, and to assess the frequency with which chest radiographs (CXRs) were utilized before a diagnosis of pneumonia or pulmonary TB was made. DESIGN: A nested case-control study within a prospective study conducted to assess factors associated with delays in the diagnosis of TB. RESULTS: Cases (n = 85; 54%) were patients who received antibiotics for non-TB diagnoses/indications prior to TB diagnosis, and controls (n = 73; 46%) were patients who had initially received TB therapy. Median health care delay for cases was 39 days vs. 15 days (P < 0.01) for controls. Median antibiotic delay was similar among all antibiotic classes. Of 54 patients who did not have a CXR at their first health care visit, 41 (79%) received empiric antibiotics, compared to 44/105 (42%) who had a CXR (P < 0.01). Only 31/54 (57%) patients initially diagnosed with CAP had a CXR at the time of diagnosis. CONCLUSION: More widespread use of CXR when diagnosing CAP should reduce delays in diagnosing TB, and the unnecessary use of antibiotics. PMID- 15832465 TI - [The 75th annual meeting of the Japanese Society for Hygiene. Niigata, Japan. March 27-30, 2005. Abstracts]. PMID- 15832464 TI - [A case of rhinal myiasis]. PMID- 15832466 TI - [Regional meeting of the Japanese Association for Thoracic Surgery. Japan. 2005. Abstracts]. PMID- 15832467 TI - [Are complementary and classical natural medicine conducive to healthy living or only for curing diseases? Specific demands for evidence-based medicine]. PMID- 15832468 TI - [Re the editorial: "Healing power--an art of natural healing of people or still only by natural therapy?"]. PMID- 15832469 TI - In the name of traceability. PMID- 15832470 TI - Novel method of closing a tracheo-oesophageal fistula using a de-epithelialized deltopectoral flap. AB - The authors describe a novel use of a de-epithelialized deltopectoral flap for the closure of tracheo-oesophageal fistulae, which have been resistant to other methods of closure. PMID- 15832471 TI - Divergent standards for consent in research. PMID- 15832472 TI - Human tissue bill. PMID- 15832473 TI - The medical profession, patients and society: a situation report from Hungary. PMID- 15832474 TI - No need for specialist children's RECs? PMID- 15832475 TI - Arrested--for showing compassion? PMID- 15832476 TI - European decisions:--respect for parental refusal of consent, and--privacy and a genomic database. PMID- 15832477 TI - Ethics and bioterrorism research. PMID- 15832478 TI - Code of practice on access to personal information. PMID- 15832479 TI - Bioethics Convention protocol on research. PMID- 15832480 TI - The ethics of medical research: a Christian view. PMID- 15832481 TI - Does a fetus have the right to life? PMID- 15832482 TI - Corticosteroid-induced osteoporosis: a guide to optimum management. AB - Corticosteroid-induced osteoporosis is the leading cause of secondary osteoporosis and a significant cause of morbidity in both men and women. Long term use of even low-dose corticosteroids has been associated with increased risk of bone loss. Recent large randomized controlled trials have generated new knowledge on treatment strategies for patients with corticosteroid-induced osteoporosis. However, the majority of individuals receiving corticosteroids are not receiving prophylaxis for osteoporosis. Calcium and vitamin D should be recommended to patients initiating therapy with corticosteroids (and should be adequate for those receiving corticosteroids for less than 3 months). For those receiving corticosteroids for greater than 3 months, bisphosphonates are the therapy of choice, with both alendronate (alendronic acid) and risedronate (risedronic acid) approved by the US FDA for use in this indication. Calcitonin can be considered a second-line agent and should be reserved for patients who are intolerant of bisphosphonates or who are experiencing pain from a vertebral fracture. Hormone replacement therapy or testosterone therapy may be offered to those individuals on long-term corticosteroid treatment who are hypogonadal. Teriparatide (recombinant human parathyroid hormone 1-34) shows promise as a future anabolic agent for the prevention and treatment of patients with corticosteroid-induced osteoporosis. PMID- 15832483 TI - Gonadotropin-releasing hormone antagonists: pharmacology and clinical use in women. AB - Gonadotropin-releasing hormone (GnRH) antagonists are now widely used in protocols of patients with controlled ovarian hyperstimulation to treat infertility. By competitively binding to the pituitary GnRH receptor, they lead to a rapid suppression of gonadotropins and consecutively sex hormones. In the past, GnRH agonists have been exclusively used for these patients, with the disadvantage of an initial rise of gonadotropins--the flare-up effect. Several trials comparing the agonistic and antagonistic analogs of GnRH found no significant differences in oocyte quality, fertilization and pregnancy rates. Slightly lower implantation and pregnancy rates, and estradiol levels, in patients treated with GnRH antagonists has raised concern about eventual extrapituitary adverse effects. However, no convincing evidence has yet been found for any detrimental ovarian effects of GnRH antagonists. The lower rate of ovarian hyperstimulation syndrome, a potentially severe disadvantage of infertility treatment, is a positive feature of GnRH antagonists. The key point is that GnRH antagonists have been proven to be as effective and safe as GnRH agonists. This broadens the spectrum of indications for GnRH antagonists to sex hormone-dependent disorders like endometriosis, uterine fibroids, and gynecological cancers such as breast and ovarian cancer. PMID- 15832484 TI - Isoflavones and postmenopausal women: a critical review. AB - Isoflavonic phytoestrogens, or isoflavones, constitute a class of phytoestrogens that have properties similar to selective estrogen receptor modulators, and have attracted a substantial degree of attention in recent years, particularly as a possible alternative to the conventional hormone replacement therapy regimens used by postmenopausal women. Despite great promise, it is difficult to make many specific recommendations about their use at the current time, in light of the many outstanding questions that hopefully will be answered in the future by focused interventional studies involving humans. Studies to date indicate that the use of isoflavones to address vasomotor symptoms provides at most small benefits beyond a placebo effect, and no benefit for genital atrophy. As for postmenopausal women whose primary concern is cardiovascular disease, the recommendation of the American Heart Association to include soy protein foods as part of an otherwise healthy diet is well justified, and similarly the substitution of supplements containing soy protein for animal protein can also be recommended. The use of purified isoflavone supplements not containing soy protein may have some cardiovascular benefits, but these appear to be less substantial in degree than those provided by soy protein with isoflavones. In particular, more research is needed to assess the effects of isoflavones on osteoporosis, for which no recommendation regarding isoflavones can be made based on the current data. Also, isoflavones should not be taken by postmenopausal women for the specific purpose of decreasing their risk of breast or endometrial cancer, although, at least for those without pre-existing disease or at high risk, it seems quite unlikely that isoflavone use is harmful in this regard. PMID- 15832485 TI - Vitamin D analogs: new therapeutic agents for secondary hyperparathyroidism. AB - Patients with chronic renal failure frequently develop secondary hyperparathyroidism, primarily as a result of phosphate retention and low serum 1,25(OH)2D3. Replacement therapy with calcitriol or its synthetic precursor alfacalcidol [1alpha(OH)D3] often produces hypercalcemia, especially when combined with calcium-based phosphate binders. In addition, the natural vitamin D compounds can exacerbate the hyperphosphatemia in patients with chronic renal failure. This combined increase in calcium and phosphate has been correlated with vascular calcification leading to coronary artery disease, the most common cause of mortality in renal patients. Several vitamin D analogs have now been developed that retain the direct suppressive action of calcitriol on the parathyroid glands but have less calcemic activity, thereby offering a safer and more effective means of controlling secondary hyperparathyroidism. Maxacalcitol [22-oxa 1,25(OH)2D3] and falecalcitriol [1,25(OH)2-26,27-F6-D3] are currently available in Japan, and paricalcitol [19-nor-1,25(OH)2D2] and doxercalciferol [1alpha(OH)D2] are available in the US. The mechanisms by which these analogs exert their selective actions on the parathyroid glands are under investigation. The low calcemic activity of maxacalcitol has been attributed to its rapid clearance from the circulation. This prevents sustained effects on intestinal calcium absorption and bone resorption, but still allows a prolonged suppression of parathyroid hormone gene expression. The selectivity of the other analogs is achieved by distinct mechanisms. Understanding how these compounds exert their selective actions on the parathyroid glands will aid in the design of safer, more effective analogs. PMID- 15832486 TI - Improved glycemic control and lipid profile in a randomized study of pioglitazone compared with acarbose in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To assess the efficacy of pioglitazone treatment in comparison with that of acarbose treatment in patients with type 2 diabetes mellitus. PARTICIPANTS AND METHODS: In this randomized, parallel-group, open-label study patients were assigned to treatment with either pioglitazone (n = 129) or acarbose (n = 136). During a 1-week run-in patients commenced an individualized dietary regimen which was maintained throughout the study. Patients received the assigned study medication for 26 weeks. Serum glycosylated hemoglobin (HbA1c) levels, insulin resistance and lipid profiles were determined at baseline and at endpoint. RESULTS: Mean HbA1c was reduced from 8.98+/-1.20% to 7.82+/-1.95% with pioglitazone treatment and from 9.03+/-1.32% to 8.55+/-1.96% with acarbose treatment during the 26-week study. The change from baseline to endpoint was significantly greater for pioglitazone compared with acarbose when analyzed for all patients (p < 0.001) and for those who had (p = 0.009) or had not (p < 0.001) received previous medication for diabetes mellitus. Compared with acarbose, pioglitazone produced a significantly greater decrease in fasting glucose, insulin and insulin resistance (p < 0.001 for each). Triglycerides were decreased by 71.1+/-184.1 mg/dl with pioglitazone compared with 38.1+/-171.3 mg/dl with acarbose (p = 0.001 for difference between groups). High density lipoprotein (HDL)-cholesterol level was increased by 7.8+/-10.2 mg/dl with pioglitazone compared with a decrease of 0.8+/-24.1 mg/dl with acarbose (p < 0.001). While serum low density lipoprotein (LDL)-cholesterol levels remained unchanged with both treatment regimens, the decrease from baseline in very low density lipoprotein (VLDL)-cholesterol was significantly greater with pioglitazone than with acarbose (p < 0.04). Pioglitazone decreased systolic blood pressure by 5.6+/ 17.7mm Hg compared with a 0.4+/-18.4mm Hg increase during acarbose treatment (p < 0.001). Pioglitazone caused a significantly greater decrease compared with acarbose in serum levels of gamma-glutamyl aminotransferase (p < 0.001) and alanine aminotransferase (p = 0.004). CONCLUSIONS: Six months of pioglitazone treatment decreased insulin resistance and improved glycemic control to a significantly greater extent than acarbose treatment. Pioglitazone was also associated with a significantly improved lipid profile, suggesting a reduction in risk of coronary heart disease. PMID- 15832487 TI - Opinion and evidence for treatments in endocrine disorders. AB - New treatments and treatment protocols for endocrine disorders are evolving rapidly, and research and development activity in the endocrinology field is high. Optimal therapy remains contentious in some areas. To help you keep up-to date with the latest advances worldwide on all aspects of drug therapy and management of endocrine disorders, this section of the journal brings you information selected from the rapid drug news alerting service Inpharma Weekly. Each issue contains easy-to-read summaries of the most important research and development news, clinical studies, treatment guidelines, pharmacoeconomic and adverse drug reaction news, and expert opinion pieces published in the world's top endocrinology journals. PMID- 15832488 TI - Effects of dehydroepiandrosterone on bone mineral density: what implications for therapy? AB - Because dehydroepiandrosterone (DHEA) levels decline dramatically with aging and low DHEA levels correlate with age-related diseases, it has been suggested that old age may represent a condition of DHEA deficiency. Accordingly, there have been some studies of the effects of restoring the DHEA levels of older individuals back to the normal range in the young. Emerging evidence from these studies shows that prasterone (DHEA replacement) may significantly enhance bone mineral density (BMD). In fact, the improvements of BMD in response to prasterone are accompanied not only by suppression of bone resorption but more importantly, stimulation of bone formation. Thus, prasterone appears to have additional anabolic effects on the skeleton, which represents an advantage over current pharmacologic agents that only inhibit bone loss. The osteogenic effects in elderly people are consistent with DHEA serving primarily as a precursor to active androgens and estrogens in local tissues such as bone. DHEA replacement may also increase levels of insulin-like growth factor-1, which may contribute to its anabolic effects. Although prasterone may be an effective therapy for improving BMD in both sexes, there appears to be gender differences in responses with more osteogenic effects in older women compared with older men. More studies, particularly randomized, placebo-controlled trials which include fractures as an outcome, are needed to fully define the potential utility of DHEA replacement as an anabolic intervention for age-related osteoporosis. These studies would also be important to gain information on risks associated with long term DHEA replacement therapy. Further investigations are particularly warranted because prasterone is available over-the-counter in many countries and many older individuals are taking this hormone, without medical supervision, for its purported anti-aging properties. PMID- 15832489 TI - Pathophysiology of type 2 diabetes mellitus in children and adolescents: treatment implications. AB - Type 2 diabetes mellitus was considered an exclusive disease of adulthood until the late 1970s, when reports of an increased prevalence in the pediatric age group emerged in the literature. The concerning upswing in the rate of diagnosis of type 2 diabetes mellitus in children and adolescents has continued, parallel to the increasing rates of obesity. The disease is not specific to the U.S.; it has proven to be a global problem. The current information on type 2 diabetes mellitus in children and adolescents is mostly extrapolated from studies in adults with type 2 diabetes mellitus, due to the paucity of studies conducted in youth. Obesity, family history of type 2 diabetes mellitus, minority ethnicity and race, polycystic ovary syndrome, maternal diabetes mellitus or impaired glucose tolerance during gestation, and acanthosis nigricans are the major risk factors and markers of youth-onset type 2 diabetes mellitus. The pathophysiology, which involves both an insulin secretion defect and resistance to insulin, needs further clarification in pediatric studies. Current management approaches involve lifestyle modification (nutritional and exercise) along with pharmacologic agents, such as insulin and oral antihyperglycemic medications, as indicated. A recent study on the use of metformin in childhood-onset type 2 diabetes mellitus demonstrated the drug to be effective and to have a good safety profile in this population. However, the outcomes of ongoing studies and future studies focusing on type 2 diabetes mellitus in the pediatric age group will be crucial in terms of fine-tuning management plans and setting up appropriate prevention strategies. PMID- 15832490 TI - Managing cutaneous manifestations of hyperandrogenic disorders: the role of oral contraceptives. AB - Cutaneous manifestations of hyperandrogenic disorders (acne, seborrhea, hirsutism and androgenetic alopecia) can be caused by elevated levels of free testosterone or androgen precursors. In women with normal serum levels of testosterone or androgen precursors, enhanced local conversion to testosterone, or to the more potent androgen dihydrotestosterone, may lead to increased androgen activity in the pilosebaceous unit. Large individual variations in the response to normal or elevated androgens suggests considerable differences in local androgen metabolism and androgen receptor-mediated activities, which may partly be related to genetic disposition. Androgens cause opposite effects on hair follicles in the scalp compared with the face and body, and there are large differences in the length of anagen phase. Androgens enhance sebum production and keratinization, prolong the growth phase of face and body hair, stimulate the transformation of vellus to terminal hair, and shorten the anagen phase of scalp hair. Estrogens may antagonize the androgen-induced actions on sebaceous glands and hair follicles. Treatment with oral contraceptives (OCs) reduces the production of androgens and androgen precursors and increases sex hormone-binding globulin, resulting in a decrease of free testosterone levels. According to type and dose, the estrogen and progestogen components of OCs may directly reduce the effect of androgens within sebaceous glands and hair follicles. Therefore, OCs with a predominant estrogen effect may improve mild to moderate forms of acne and seborrhea, hirsutism and androgenetic alopecia, in a time-dependent manner. In women who do not respond satisfactorily, treatment with OCs containing a progestogen with antiandrogenic activity is recommended. In many women with severe acne or hirsutism, a considerable increase in the local concentration of the antiandrogenic progestogen is required to reduce the androgenic interaction with the androgen receptor. For this therapy, an OC containing cyproterone acetate can be used. If necessary, the dose of cyproterone acetate can be increased in a stepwise manner. While androgenetic alopecia is best treated with a low-dose OC containing cyproterone acetate (optimal effect occurs after at least 12 months of therapy), severe acne and hirsutism are significantly improved after 6-12 months of regimens containing high doses of cyproterone acetate (25-100 mg/day). After termination of treatment the disorders may reappear, therefore treatment with suitable low-dose formulations is recommended to maintain the therapeutic effect. PMID- 15832491 TI - Management of obesity in the elderly: special considerations. AB - Over the last few decades, there has been an unprecedented increase in the prevalence of obesity, especially in economically developed countries. Furthermore, it is becoming an increasingly recognized health problem in the elderly. The precise mechanisms underlying increased adiposity in the elderly are not known. Aging is associated with a host of biologic changes that limit the ability of the individual to regulate energy homeostasis. Thus, it is likely that older individuals may be more likely to develop the two extremes of the spectrum of nutritional abnormalities, namely malnutrition and increased adiposity. These nutritional abnormalities are associated with significant morbidity and mortality. Current guidelines define overweight as a body mass index (BMI) of 25 29.9 kg/m2 and obesity as a BMI of 30 kg/m2 or more. However, the optimal BMI may be different in older individuals. Management strategies should attempt to optimize the nutritional status of older individuals. Age per se cannot be used as a justification for denying medical management of obesity to elderly individuals. Individualized programs with the goal of achieving modest weight reduction in obese patients are likely to result in immediate (e.g. alleviation of arthritic pains and reduction of glucose intolerance) and possibly long-term (e.g. reduction in cardiovascular risk) healthcare benefits. Management should emphasize lifestyle modifications, while the use of pharmacologic agents such as sibutramine and orlistat should be reserved for select groups of patients who do not respond to lifestyle modification. PMID- 15832492 TI - Therapeutic potential of insulin-like growth factor-1 in patients with diabetes mellitus. AB - Insulin-like growth factor-1 (IGF-1) and its receptors share considerable homology with insulin and insulin receptors, and their respective signaling pathways interact at the post receptor level. While the growth hormone (GH)-IGF-1 axis principally regulates tissue growth and differentiation, insulin exerts it primary effects on fuel metabolism. However, these two endocrine systems interact at multiple levels and in diabetes mellitus the GH-IGF-1 axis is grossly disturbed, with increased secretion of GH, reduced plasma levels of IGF-1, and complex tissue-specific changes in IGF binding proteins (IGFBPs). These observations have given rise to the view that GH-IGF-1 axis dysfunction, particularly low plasma levels of circulating IGF-1, probably play a significant role in several aspects of the pathophysiology of diabetes mellitus, including insulin resistance and poor glycemic control, and may also influence the development of microvascular complications. The availability of recombinant human IGF-1 (rhIGF-1; mecasermin), used either alone or in combination with insulin, has led to experimental studies and clinical trials in humans testing these hypotheses. These studies have examined the impact of subcutaneous rhIGF-1 injections on sensitivity and metabolic parameters. In patients with type 1 and 2 diabetes mellitus, insulin sensitivity is significantly improved, insulin requirements are reduced, and glycemic control of dyslipidemia is generally improved in short-term studies. rhIGF-1 is a particularly attractive possibility in patients with type 2 diabetes mellitus, where insulin resistance is the fundamental problem. Some patients with genetic syndromes of severe insulin resistance also benefit from treatment with rhIGF-1, which can bypass blocks in the insulin signaling pathway. The common adverse effects reported for rhIGF-1 are dose-related and include edema, jaw pain, arthralgia, myalgia, hypotension, injection site pain, and less commonly, Bell's palsy and raised intracranial pressure. Although disturbance of the GH-IGF-1 axis participates in the development of diabetic complications, the functional consequences of the complex changes in IGFBP expression at the tissue level are uncertain, and it is not known whether systemic IGF-1 therapy or other manipulations of the GH-IGF-1 axis would be helpful or harmful. Experimentally, IGF-1 has a protective effect on neuropathy, and could find an application in the healing of neuropathic ulcers. The potential benefits of IGF-1 therapy in diabetes mellitus have yet to be realised. PMID- 15832493 TI - Spotlight on rosiglitazone in the management of type 2 diabetes mellitus. AB - Rosiglitazone, a thiazolidinedione with a different side chain from those of troglitazone and pioglitazone, reduces plasma glucose levels and glucose production and increases glucose clearance in patients with type 2 diabetes mellitus. Insulin sensitivity, pancreatic beta-cell function and surrogate markers of cardiovascular risk factors are significantly improved by rosiglitazone. Double-blind trials of 8 to 26 weeks of rosiglitazone 4 or 8 mg/day monotherapy indicate significant decreases in fasting plasma glucose (-2 to -3 mmol/L with 8 mg/day) and glycosylated hemoglobin levels [HbA(1c); -0.6 to 0.7% (-0.8 to -1.1% in drug-naive patients) with 8 mg/day]. Significant decreases in hyperglycemic markers occurred when rosiglitazone was combined with metformin (HbA(1c) -0.8 to -1.0%), a sulfonylurea (-1.4%) or insulin (-1.2%) for 26 weeks versus little change with active comparator monotherapy. Efficacy was maintained in trials of < or =2 years, and was also apparent in various ethnic subgroups, elderly patients and both obese and nonobese patients. Rosiglitazone is currently not indicated in combination with injected insulin. It should be administered in conjunction with diet and exercise regimens. Rosiglitazone is generally well tolerated. Despite rare individual reports of liver function abnormalities in rosiglitazone recipients, the incidence of these in clinical trials (< or =2 years' duration) was similar to that in placebo and active comparator groups. Fluid retention associated with rosiglitazone may be the cause of the increased incidence of anemia in clinical trials, and also means that patients should be monitored for signs of heart failure during therapy. Although bodyweight is increased overall with rosiglitazone therapy, increases are in subcutaneous, not visceral, fat; hepatic fat is decreased. The pharmacokinetic profile of rosiglitazone is not substantially altered by age or renal impairment, nor are there important drug interactions. Rosiglitazone is not indicated in patients with active liver disease or increased liver enzymes. CONCLUSIONS: Oral rosiglitazone 4 or 8 mg/day provides significant antihyperglycemic efficacy and is generally well tolerated, both as monotherapy and in combination with other antihyperglycemic agents, in patients with type 2 diabetes mellitus who do not have active liver disease. Long-term data are required before conclusions can be drawn about the clinical significance of positive changes to surrogate markers of cardiovascular disease risk and improvements to pancreatic beta-cell function. Rosiglitazone significantly improves insulin sensitivity and, as such, is a welcome addition to the treatment options for patients with type 2 diabetes mellitus. PMID- 15832494 TI - Opinion and evidence for treatments in endocrine disorders. AB - New treatments and treatment protocols for endocrine disorders are evolving rapidly, and research and development activity in the endocrinology field is high. Optimal therapy remains contentious in some areas. To help you keep up-to date with the latest advances worldwide on all aspects of drug therapy and management of endocrine disorders, this section of the journal brings you information selected from the rapid drug news alerting service Inpharma Weekly. Each issue contains easy-to-read summaries of the most important research and development news, clinical studies, treatment guidelines, pharmacoeconomic and adverse drug reaction news, and expert opinion pieces published in the world's top endocrinology journals. PMID- 15832495 TI - Finding the "Magic". PMID- 15832496 TI - Cytosolic delivery of viral nucleoprotein by listeriolysin O-liposome induces enhanced specific cytotoxic T lymphocyte response and protective immunity. AB - Cytotoxic T lymphocytes (CTLs) are capable of conferring protection against intracellular pathogens and tumor. Protective antiviral immunity, mediated by the activation of antigenic epitope-specific CTL, can be achieved by delivering exogenous antigen into the cytosol of antigen-presenting cells. Cytosolic introduction of vaccine antigen, however, requires a specialized delivery strategy due to the membrane barrier limiting the access of macromolecules to the cytosol. In this study, we have investigated the potential ability of listeriolysin O-containing liposomes (LLO-liposomes) to deliver lymphocytic choriomeningitis virus (LCMV) nucleoprotein (NP), harnessing the intracellular invasion mechanism of Listeria monocytogenes, to stimulate a NP-specific CTL response. We have analyzed the ability of LLO-liposomes to induce an enhanced CTL response and determined the extent of CTL-mediated protection using an in vivo infection model. Mice immunized with LLO-liposomes containing NP generated a higher frequency of NP-specific CD8+ T cells with greater effector activity than the control groups immunized with either non-LLO-liposomal NP or LLO-liposomes containing control protein. Moreover, LLO-liposomal NP-immunized mice were completely protected against a lethal intracerebral challenge with a virulent strain of LCMV and were capable of clearing a chronic LCMV infection. Our study demonstrates that LLO-liposomes can be used as an efficient vaccine delivery system carrying a viral antigenic protein to generate protective antiviral immunity. PMID- 15832497 TI - Novel antitumor prodrugs designed for activation by matrix metalloproteinases-2 and -9. AB - Enzyme prodrug monotherapy takes advantage of the selectivity and specificity of enzymes that are differentially active in the immediate environment of tumor cells. Matrix metalloproteinases-2 and -9 (MMP-2 and -9, respectively) are cell surface Zn-dependent endoproteases associated with diverse processes throughout tumor formation and progression. These enzymes have demonstrated high ratios of tumor- to nontumor-associated activity and may represent candidates for antitumor prodrug activation. Our MMP targeting strategy was to prepare and evaluate two classes of enzyme prodrugs, peptides and sequence-similar peptidomimetics, and determine which would be substrates for the enzymes and thus suitable for further in vitro and in vivo evaluation. We selected representatives of three structurally and mechanistically distinct classes of compounds for delivery, doxorubicin, several auristatins (novel synthetic members of the dolastatin class of tubulin polymerization inhibitors), and CBI-TMI (a duocarmycin class minor groove binder). The drugs were acylated on available amines with the broadly recognized MMP substrate P3-P1' sequence acetyl L-prolyl-L-leucyl-glycyl-L leucine, or with a peptidomimetic analogue. From a panel of four peptides and four peptidomimetics, two compounds, both peptides, were found to be substrates, with specific activities in the range of 1-20 nmol min(-1) mg(-1). For MMP-9, complete conversion took place in 4-16 h; proteolysis by MMP-2 was considerably slower. Cleavage occurred, as predicted, at the Gly-Leu bond to liberate a leucyl drug, and no other intermediates or cleavage products were observed. Although the MMP-9 proteolysis products were equipotent with the parent leucyl drugs, the prodrugs were not differentially active against MMP-2 or -9-expressing versus nonexpressing cell lines during a 4 h exposure. Our data can be interpreted in light of the current understanding of the structural and mechanistic factors governing MMP-2 and -9 proteolysis. PMID- 15832498 TI - GSSI, a general model for solute-solvent interactions. 1. Description of the model. AB - A novel, semiempirical approach for the general treatment of solute-solvent interactions (GSSI) was developed to enable the prediction of solution-phase properties (e.g., free energies of desolvation, partition coefficients, and membrane permeabilities). The GSSI approach is based on the principle that all solution-phase processes can be modeled in terms of one or more gas-to-solution transfer processes. The free energy of each gas-to-solution transfer process is calculated as the sum of the free energy of cavity formation and the free energy of solute-solvent interaction. The solute's contributions to these free energies are modeled on the basis of various quantities computed from the solute's three dimensional (3D) structure, whereas the solvent's contributions are modeled by empirically determined regression coefficients. More specifically, the free energy of cavity formation is modeled on the basis of the total solvent accessible surface area of the solute. The enthalpy of solute-solvent interaction is modeled on the basis of intermolecular interaction potentials calculated at many uniformly distributed points on the solvent-accessible surface of the solute. The entropy of solute-solvent interaction is modeled on the basis of an effective number of rotatable bonds in the solute and by the regression coefficients characteristic of the solvent. The potential utility of the GSSI approach was demonstrated by modeling the free energy of gas-to-solution transfer for 111 solutes in water, 250 solutes in hexadecane, and 84 solutes in octanol. PMID- 15832499 TI - Increased acyclovir oral bioavailability via a bile acid conjugate. AB - The objective of this work was to design an acyclovir prodrug that would utilize the human apical sodium-dependent bile acid transporter (hASBT) and enhance acyclovir oral bioavailability. Using each chenodeoxycholate, deoxycholate, cholate, and ursodeoxycholate, four bile acid prodrugs of acyclovir were synthesized, where acyclovir was conjugated to a bile acid via a valine linker. The affinity of the prodrug for hASBT was determined through inhibition of taurocholate uptake by COS-7 cells transfected with hASBT (hASBT-COS). The prodrug with the highest inhibitory affinity was further evaluated in vitro and in vivo. The prodrug acyclovir valylchenodeoxycholate yielded the highest affinity for hASBT (Ki = 35 microM), showing that chenodeoxycholate is the free bile acid with the greatest affinity for hASBT. Acyclovir valylchenodeoxycholate's affinity was similar to that of cholic acid (Ki = 25 microM). Further characterization showed that acyclovir was catalytically liberated from acyclovir valylchenode-oxycholate by esterase. Relative to cellular uptake studies of acyclovir alone, the cellular uptake from the prodrug resulted in a 16-fold greater acyclovir accumulation within hASBT-COS cells, indicating enhanced permeation properties of the prodrug. Enhanced permeability was due to hASBT-mediated uptake and increased passive permeability. The extent of acyclovir uptake in the presence of sodium was 1.4-fold greater than the extent of passive prodrug uptake in the absence of sodium (p = 0.02), indicating translocation of the prodrug by hASBT. The prodrug also exhibited an almost 12 fold enhanced passive permeability, relative to acyclovir's passive permeability. Oral administration of acyclovir valylchenodeoxycholate to rats resulted in a 2 fold increase in the bioavailability of acyclovir, compared to the bioavailability after administration of acyclovir alone. Results indicate that a bile acid prodrug strategy may be useful in improving the oral bioavailability of intestinal permeability-limited compounds. PMID- 15832500 TI - Human organic anion-transporting polypeptide OATP-A (SLC21A3) acts in concert with P-glycoprotein and multidrug resistance protein 2 in the vectorial transport of Saquinavir in Hep G2 cells. AB - Saquinavir mesylate (SQV) is the first-in-class and prototypical HIV protease inhibitor (PI) used in the treatment of HIV infection. SQV undergoes extensive hepatic metabolism and intestinal and bile secretion, and has poor and variable oral bioavailability. In previous studies, our group and others have described the interactions between SQV and absorptive and secretory efflux transporters such as MRP1, MRP2, and P-gp. However, the potential role of absorptive influx transporters such as OATP-A (SLC21A3) has not yet been reported for SQV. In the study presented here, the role of OATP-A in the influx transport of SQV was studied using a hepatic cell model, Hep G2, and Xenopus laevis oocytes overexpressing human OATP-A. In Hep G2 cells, SQV transport was found to be (i) concentration-dependent and saturable, (ii) temperature-sensitive, and (iii) proton (pH)- and sodium-independent. While GF120918, a specific inhibitor of P gp, and MK571, a MRP transporter family inhibitor, significantly enhanced SQV uptake, estrone 3-sulfate, a substrate of OATP-A, significantly inhibited SQV uptake by Hep G2 cells. The observation that inhibitors of P-gp, MRP, or OATP-A have opposite effects on SQV uptake in polarized Hep G2 cells is consistent with their functions as hepatic efflux or influx transporters. In X. laevis oocytes into which OATP-A cRNA had been injected, the level of uptake of SQV was significantly greater than the level of uptake by oocytes into which water had been injected and was concentration-dependent and saturable (Km = 36.4+/-21.8 microM). This is the first report showing that SQV influx transport is directly facilitated by OATP-A. Given the wide body distribution of OATP-A, the current results suggest a potentially important role for OATP-A in the absorption and disposition of SQV in vivo. The data also suggest that in human hepatocytes basolaterally located OATP-A (influx transporter) may act in concert with apically located P-gp and/or MRP2 (efflux transporters) for the vectorial transport and excretion of SQV into bile. PMID- 15832501 TI - Involvement of OCTN1 (SLC22A4) in pH-dependent transport of organic cations. AB - OCTN1 (SLC22A4) transports cationic compounds such as tetraethylammonium in a pH sensitive and sodium-independent manner in cultured cells, and is expressed in wide variety of tissues, including kidney, muscle, placenta, heart, and others. This study focused on the clarification of its subcellular distribution in kidney and on its driving force to throw light on the pharmacological and physiological roles of OCTN1. Uptake of [14C]tetraethylammonium by membrane vesicles prepared from HEK293 cells stably transfected with human OCTN1 cDNA was osmolarity sensitive, and the Km of tetraethylammonium was 1.28 mM at intravesicular and extravesicular pH values of 6.0 and 7.4, respectively. Tetraethylammonium uptake was pH-dependent, and overshoot uptake was observed in the presence of an outwardly directed proton gradient. A protonophore and membrane potential affected the overshoot uptake. Furthermore, preloading tetraethylammonium in the vesicles significantly increased the rate of uptake of [14C]tetraethylammonium. In mouse kidney, OCTN1 was expressed predominantly at the apical membrane of cortical proximal tubular epithelial cells. It was concluded that OCTN1 is involved in renal excretion of organic cations across the apical membrane in a pH dependent, membrane potential-sensitive manner and is affected significantly by the organic cations on the trans side, showing counter transport activity. PMID- 15832502 TI - A novel high-throughput pepT1 transporter assay differentiates between substrates and antagonists. AB - PepT1 is a transporter of proven pharmaceutical utility for enhancing oral absorption. A high-throughput, robust functional assay, capable of distinguishing PepT1 binders from substrates, allowing identification and/or prediction of drug candidate activation was developed. An MDCK epithelial cell line was transfected with rPepT1. The high level of stable rPepT1 expression that was achieved enabled development of a miniaturized PepT1 assay in a 96-well format, which could be scaled to 384 wells. The assay is based on measurement of membrane depolarization resulting from the cotransport of protons and PepT1 substrates. Membrane potential changes are tracked with a voltage-sensitive fluorescent indicator. Control (mock-transfected) cells are used to determine nonspecific membrane potential changes. A variety of fluorescent dyes were tested during initial assay design, including intracellular pH and membrane potential indicators. A membrane potential indicator was chosen because of its superior performance. Upon PepT1 activation with glycylsarcosine, dose-dependent membrane depolarization was observed with an EC50 of 0.49 mM. Maximum depolarization was dependent on the level of PepT1 expression. Testing of 38 known PepT1 substrates, binders, and nonbinders demonstrated that this assay accurately distinguished substrates from binders and from nonbinders. Initial validation of this novel assay indicates that it is sensitive and robust, and can distinguish between transporter substrates and antagonists. This important distinction has been previously achieved only with lower-throughput assays. This assay might also be used to determine substrate potency and establish a high-quality data set for PepT1 SAR modeling. PMID- 15832503 TI - Sustained cytoplasmic delivery of drugs with intracellular receptors using biodegradable nanoparticles. AB - Efficient cytoplasmic delivery of therapeutic agents is especially important for drugs with an intracellular site of action for elicitation of a maximal therapeutic effect. In this study, we demonstrate the efficacy of biodegradable nanoparticles for cytoplasmic delivery of dexamethasone, a glucocorticoid, whose site of action is intracellular. Equal doses of two formulations of drug-loaded nanoparticles releasing different doses of the encapsulated drug were tested for antiproliferative activity in vascular smooth muscle cells. The antiproliferative activity of the drug was significantly greater and sustained with nanoparticles that released a higher dose of the drug than with nanoparticles which released a lower dose of the drug. The greater antiproliferative activity of the nanoparticles that released a higher dose of the drug correlated with sustained and higher intracellular drug levels. The antiproliferative activity of the drug in solution was lower and relatively transient compared to that with drug-loaded nanoparticles. The mechanism of inhibition of cell proliferation was mediated through inhibition of cell-cycle progression with a relatively higher percentage of cells in the G0/G1 arrest phase in the group that was treated with drug-loaded nanoparticles compared to that treated with the drug in solution. Results of the study thus suggest that the dose and duration of a drug's availability at the intracellular site of action determine its therapeutic efficacy. In conclusion, biodegradable nanoparticles could be used as an effective delivery mechanism for sustained intracellular delivery of different therapeutic agents. PMID- 15832504 TI - Molecular properties of WHO essential drugs and provisional biopharmaceutical classification. AB - The purpose of this study is to provisionally classify, based on the Biopharmaceutics Classification System (BCS), drugs in immediate-release dosage forms that appear on the World Health Organization (WHO) Essential Drug List. The classification in this report is based on the aqueous solubility of the drugs reported in commonly available reference literature and a correlation of human intestinal membrane permeability for a set of 29 reference drugs with their calculated partition coefficients. The WHO Essential Drug List consists of a total of 325 medicines and 260 drugs, of which 123 are oral drugs in immediate release (IR) products. Drugs with dose numbers less than or equal to unity [Do = (maximum dose strength/250 mL)/solubility < or = 1] are defined as high solubility drugs. Drug solubility for the uncharged, lowest-solubility form reported in the Merck Index or USP was used. Of the 123 WHO oral drugs in immediate-release dosage forms, 67% (82) were determined to be high-solubility drugs. The classification of permeability is based on correlations of human intestinal permeability of 29 reference drugs with the estimated log P or CLogP lipophilicity values. Metoprolol was chosen as the reference compound for permeability and log P or CLogP. Log P and CLogP were linearly correlated (r2 = 0.78) for 104 drugs. A total of 53 (43.1%) and 62 (50.4%) drugs on the WHO list exhibited log P and CLogP estimates, respectively, that were greater than or equal to the corresponding metoprolol value and are classified as high permeability drugs. The percentages of the drugs in immediate-release dosage forms that were classified as BCS Class 1, Class 2, Class 3, and Class 4 drugs using dose number and log Pwere as follows: 23.6% in Class 1, 17.1% in Class 2, 31.7% in Class 3, and 10.6% in Class 4. The remaining 17.1% of the drugs could not be classified because of the inability to calculate log P values because of missing fragments. The corresponding percentages in the various BCS classes with dose number and CLogP criteria were similar: 28.5% in Class 1, 19.5% in Class 2, 35.0% in Class 3, and 9.8% in Class 4. The remaining 7.3% of the drugs could not be classified since CLogP could not be calculated. These results suggest that a satisfactory bioequivalence (BE) test for more than 55% of the high-solubility Class 1 and Class 3 drug products on the WHO Essential Drug List may be based on an in vitro dissolution test. The use of more easily implemented, routinely monitored, and reliable in vitro dissolution tests can ensure the clinical performance of drug products that appear on the WHO Essential Medicines List. PMID- 15832505 TI - An azepanone-based inhibitor of human cathepsin K with improved oral bioavailability in the rat and the monkey. PMID- 15832507 TI - Synthesis and biological evaluation of a cytarabine phosphoramidate prodrug. AB - Recently, we reported a novel approach for the intracellular delivery of the anti cancer nucleotide 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP) using phosphoramidate-based prodrugs. These phosphoramidate prodrugs contain an ester group that undergoes intracellular activation, liberating phosphoramidate anion, which in turn undergoes spontaneous cyclization and P-N bond cleavage to yield the nucleoside monophosphate quantitatively. This approach has now been extended to cytarabine [1-beta-D-arabinofuranosylcytosine (Ara-C)], an anti-cancer nucleoside that is limited in its utility because of poor intracellular transport characteristics and weak activity as a substrate for tumor cell kinases. The cytarabine phosphoramidate prodrug 1 has been synthesized and evaluated in comparison with cytarabine for growth inhibitory activity against wild-type, nucleoside transport-deficient, and nucleoside kinase-deficient CEM leukemia cell lines. The prodrug was comparable in growth inhibitory activity (IC50 = 32 nM) to cytarabine (IC50 = 16 nM) in wild-type CCRF-CEM cells following drug treatment for 72 h. The nucleoside transport-deficient CEM/AraC8C exhibited a high level of resistance (6400-fold) to cytarabine but was more sensitive (210-fold resistant vs CCRF-CEM cells) to prodrug 1. Similarly, the deoxycytidine kinase-deficient cell line (CEM/dCK-) was highly resistant to cytarabine (13900-fold) but more sensitive (106-fold resistant vs CCRF-CEM cells) to prodrug 1. These results indicate that prodrug 1 is significantly more potent than cytarabine against transport- and kinase-deficient cell lines and are consistent with a mechanism involving intracellular delivery of cytarabine 5'-monophosphate. PMID- 15832506 TI - Direct measurement of nucleoside monophosphate delivery from a phosphoramidate pronucleotide by stable isotope labeling and LC-ESI(-)-MS/MS. AB - Amino acid phosphoramidates of nucleosides have been shown to be potent antiviral and anticancer agents with the potential to act as nucleoside monophosphate prodrugs. To access their ability to deliver 3'-azido-3'-deoxythymidine (AZT) 5' monophosphate to cells, the decomposition pathway of an 18O-labeled AZT amino acid phosphoramidate was investigated by capillary reverse-phase high-performance liquid chromatography interfaced with negative ion electrospray ionization mass spectrometry (LC-ESI(-)-MS/MS). 18O-labeled L-AZT tryptophan phosphoramidate methyl ester ([18O]2) was synthesized with an 18O/16O relative ratio of 1.22 +/- 0.18. For CEM cells, a human T-lymphoblast leukemia cell line, incubated with [18O]2, values of 1.55 +/- 0.37, 0.34, and 0.13 were found for the 18O/16O relative ratio of intracellular AZT-MP for time intervals of 0.5, 4, and 20 h, respectively. The decrease in the level of labeled AZT-MP in CEM cells corresponded to a rapid increase in the amount of intracellular AZT presumably by dephosphorylation of AZT-MP. In contrast, for peripheral blood mononuclear cells (PBMCs), the 18O/16O relative ratio values of intracellular AZT-MP were 1.43, 1.06, and 0.61 for time intervals of 0.5, 4, and 20 h, respectively. Intracellular AZT in PBMCs was nearly undetectable for each time interval. Taken together, these results are consistent with the detection of direct P-N bond cleavage by CEM cells and PBMCs. However, AZT phosphoramidates are able to more effectively deliver AZT-MP to PBMCs than to CEM cells. Differential expression of 5'-nucleotidase in CEM cells relative to PBMCs is likely the reason for this discrepancy. Although applied to a phosphoramidate pronucleotide, the judicious use of 18O labeling and LC-MS is a general approach that could be applied to the investigation of the intracellular fate of other pronucleotides. PMID- 15832508 TI - A novel nucleoside prodrug-activating enzyme: substrate specificity of biphenyl hydrolase-like protein. AB - Biphenyl hydrolase-like protein (BPHL, NCBI accession number NP_004323) is a novel human serine hydrolase recently identified as a human valacyclovirase, catalyzing the hydrolytic activation of the antiviral prodrugs valacyclovir and valganciclovir. The substrate specificity of BPHL was investigated with a series of amino acid ester prodrugs of the therapeutic nucleoside analogues: acyclovir, zidovudine, floxuridine, 2-bromo-5,6-dichloro-1-(beta-D-ribofuranosyl) benzimidazole, and gemcitabine. The hydrolysis of typical esterase and aminopeptidase substrates by BPHL was also investigated. The results indicate that the substrate specificity of BPHL is largely determined by the amino acid acyl promoiety, and is less sensitive to the nucleoside parent drugs. For all nucleoside parent drugs, BPHL preferred the hydrophobic amino acids valine, phenylalanine, and proline over the charged amino acids lysine and aspartic acid. The position and monoester or diester form of the prodrug were also important, with BPHL exhibiting higher affinity for the 5'-esters than for the 3'-esters and the 3',5'-diesters irrespective of amino acid type. Further, the presence of the 3'-amino acid ester considerably reduced the hydrolysis rate of the 5'-amino acid ester functionality. BPHL exhibited stereoselectivity with an L/D specificity ratio of 32 for 5'-valyl floxuridine and 1.5 for 5'-phenylalanyl floxuridine. The substrate specificity suggests that the substrate-binding pocket of BPHL has a hydrophobic acyl binding site which can accommodate the positively charged alpha amino group, while having an alcohol leaving group binding site that can accommodate nucleoside analogues with a relatively generous spatial allowance. In conclusion, BPHL catalyzes the hydrolytic activation of amino acid esters of a broad range of therapeutic nucleoside analogues in addition to valacyclovir and valganciclovir and has considerable potential for utilization as an activation target for design of antiviral and anticancer nucleoside analogue prodrugs. PMID- 15832509 TI - Rapid quantum mechanical models for the computational estimation of C-H bond dissociation energies as a measure of metabolic stability. AB - Several relatively inexpensive levels of theory are surveyed together with alternative algorithmic methods for the estimation of C-H bond dissociation energies (BDEs), such energies being useful for the prediction of metabolic stability in drug-like molecules. In particular, bond stretching potentials of several C-H bonds are computed using the AM1, PM3, HF/MIDI!, and B3LYP/MIDI! levels of electronic structure theory, and selected points are fit to Morse and parabolic potentials. BDEs computed by an AM1 fit to the Morse function show the smallest mean unsigned error in prediction (+/- 3-4 kcal/mol) over 32 diverse C-H bonds. An alternative method for correlating the AM1 parabolic force constant from a two-point unrelaxed potential provides only a slightly decreased accuracy and is computationally particularly inexpensive. Both methods should prove to be useful for the rapid in silico screening of drug-like molecules for metabolic stability to C-H bond oxidizing enzymes. PMID- 15832510 TI - A novel hPepT1 stably transfected cell line: establishing a correlation between expression and function. AB - Stably transfected MDCK/hPepT1-V5&His clonal cell lines expressing varying levels of epitope-tagged hPepT1 protein were established to quantify the relationship between transgene hPepT1 expression levels and its functional kinetics in facilitating peptide and peptide-like drug uptake and transport in vitro. The hPepT1 sequence was amplified from Caco-2 cell mRNA, inserted into the pcDNA3.1 V5&His TOPO plasmid, and transfected into MDCK cells. Transgene protein levels were quantified by Western Blot analysis utilizing a standard curve generated with a positive control protein containing a V5&His epitope. Three clones expressing different levels of the hPepT1 fusion protein (low, medium, and high) were selected for the functional characterization with [14C]Gly-Sar and [3H]carnosine. The MDCK/hPepT1 cells expressed a novel hPepT1/epitope tag protein with an apparent molecular mass of 110 kDa. The [14C]Gly-Sar uptake in the transfected cells was sodium-independent and pH-dependent, demonstrating enhanced uptake, the rate of which increased significantly from the weakly to strongly expressing hPepT1 MDCK/hPepT1 -V5&His clones as compared to the mock cell line at pH 6.0. The uptake and permeability of [14C]Gly-Sar and [3H]carnosine demonstrated a direct correlation between the hPepT1 level of expression, uptake, and transport capabilities. Molecular and functional characterization of the MDCK/hPepT1-V5&His cell line confirmed a directly proportional relationship between Vmax and Papp versus the molar levels of hPepT1 transgene expression. This stably transfected hPepT1 cell line may serve as a useful in vitro model for screening and quantifying peptide and peptide-like drug transport as a function of hPepT1 expression in drug discovery. PMID- 15832511 TI - Quantitative assessment of the cell penetrating properties of RI-Tat-9: evidence for a cell type-specific barrier at the plasma membrane of epithelial cells. AB - Penetration of epithelial cells represents the rate-determining step for the absorption of many drugs and pharmaceutical macromolecules such as proteins and nucleic acid therapeutics. While the potential of using cell-penetrating peptides (CPPs) to facilitate absorption has been increasingly recognized, the mechanism of cell penetration and the uptake into certain cells have recently been called into question due to methodological artifacts. Therefore, the objective of this study was to quantitatively assess the ability of RI-Tat-9, a proteolytically stable CPP, to penetrate epithelial cell monolayers. The permeability of RI-Tat-9 with two epithelial cell lines, Madin-Darby canine kidney (MDCK) and Caco-2 cells, was comparable to the leakiness of the respective intact monolayers. Microscopic imaging showed that fluorescence-tagged RI-Tat-9 did not enter these cells, further supporting a paracellular transport mechanism. Although insufficient data were generated in these studies to generalize the observed phenomenon, the entry of RI-Tat-9 into nonepithelial T lymphocytic MT2 cells, possibly by endocytosis, suggested that a cell type-specific barrier might exist that controlled uptake of RI-Tat-9 by cells. Compared to that in MT2 and HeLa cells, the active uptake of the peptide into MDCK monolayers was much slower and showed no dependence of cell energy. Furthermore, the equilibrium binding of RI Tat-9 to MDCK cells at 0 degrees C was indicative of an interaction with a nonspecific receptor. A correlation between binding density and concentration difference across a leaky separation barrier suggested that repulsion of free peptide molecules by bound peptide molecules at the MDCK monolayer surface may be significant at micromolar concentrations. The results of this study quantitatively show that Tat CPP uptake into two commonly used epithelial cell types is minimal and possibly cell type-specific. Implications for Tat CPP assisted drug delivery are discussed. PMID- 15832512 TI - Bimodal effect of hypoxia in cancer: role of hypoxia inducible factor in apoptosis. AB - The effects of the separate and combined application of hypoxia and antisense oligonucleotides (ASO) against hypoxia inducible factor 1alpha (HIF1A) on cancer cells were examined. Experiments were carried out on human ovarian carcinoma cells in four series: (1) control [Normoxia (5% CO2 in air), no treatment], (2) hypoxia (1% O2, 5% CO2, and 94% N2 for 48 h), (3) treatment with ASO targeted to HIF1A (48 h), and (4) combined action of hypoxia and ASO. After treatment, the following processes and factors were monitored: apoptosis, cellular metabolism and viability, expression of genes encoding HIF1A, von Hippel-Lindau tumor suppressor protein (VHL), and genes responsible for cell death induction and antiapoptotic defense (P53, BCL2, BAX, and caspases 9 and 3). Expression of caspase 9 and HIF1A protein was confirmed by Western blotting. Liposomes were used as a delivery system of HIF1A ASO. It was found that hypoxia alone significantly disturbed cellular metabolism, reducing the level of respiration by 50% when compared with control. Hypoxia induced apoptosis by upregulating the P53 , BAX-, and caspase-dependent cell death pathways, while activating cellular antiapoptotic defense by the overexpression of BCL2 protein. Both opposing effects were dependent on the overexpression of hypoxia inducible factor. We conclude that hypoxia induces a bimodal effect, simultaneously promoting cell death and activating cellular resistance. The downregulation of HIF1A promoted cell death induction and prevented activation of cellular defense by hypoxia. This suggests that HIF1A is a potential candidate for anticancer therapeutic targeting. PMID- 15832513 TI - Solution structures of 1:1 complexes of oxyphenonium bromide with beta- and gamma cyclodextrins. AB - The solution structures of complexes of oxyphenonium bromide (OB) with beta- and gamma-cyclodextrins (beta- and gamma-CDs, respectively) in deuterium oxide have been investigated by 500 MHz proton NMR spectroscopy and molecular mechanics calculations. The chemical shifts induced by complex formation provide the 1:1 binding constants and the chemical shift variations, DeltadeltaOB-CD, with complexation for the protons of OB and the CDs. The observed binding constants are very close to those obtained by other methods and are in the following order: beta-CD > gamma-CD > alpha-CD. Initial structures of the complexes are constructed on the basis of the ROESY spectra and the DeltadeltaOB-CD values and are optimized by molecular mechanics calculations. The intermolecular distances between the protons of OB and CD calculated for these structures are well correlated with the observed ROESY intensities. The cyclohexyl group of OB penetrates deeply into a beta-CD cavity, and the phenyl group is close to the wide rim of the cavity. The phenyl and cyclohexyl groups of OB are both incorporated into a gamma-CD cavity. Furthermore, these structures of the complexes are consistent with the suppression of bitter taste and basic hydrolysis of OB by CDs and the polarity of binding sites of OB. PMID- 15832514 TI - Reducing the cardiovascular risk in diabetes. PMID- 15832515 TI - Congenital heart disease: a GP guide. PMID- 15832516 TI - Casebook: palpitations. PMID- 15832517 TI - Chronic heart failure: a review. PMID- 15832518 TI - Surgery for aortic valve disease. PMID- 15832519 TI - Comorbidity in general practice. PMID- 15832520 TI - Vaccine advice for travellers. PMID- 15832521 TI - Membership by assessment of performance. PMID- 15832522 TI - Ten per cent joy--or misery? PMID- 15832523 TI - Speech, language and literacy skills 3 years later: a follow-up study of early phonological and metaphonological intervention. AB - BACKGROUND: Three years before the present study, 19 preschool children participated in a phonological and metaphonological intervention programme. The phonological intervention programme was based on non-linear phonological analyses. The metaphonological intervention programme included both rhyming and alliteration tasks and was directly targeted during the last section of the programme. All children made significant gains in phonology, and many in metaphonology. However, the literature notes a tendency for children with history of early speech or language impairments to have difficulty acquiring literacy skills or to show residual speech impairment. The participants of the 1998 study were therefore considered at risk for continuing speech impairment and/or academic performance. AIMS: The primary objectives were to document the children's later speech, language and literacy skills, and to determine potential relationships between previous and concurrent child factors. METHODS & PROCEDURES: Twelve children from the original cohort (aged 6;1-8;5) received a comprehensive battery of speech, language, cognitive and academic tasks 3 years after the original study. Data were evaluated both concurrently and in terms of the children's preschool performance. OUTCOMES & RESULTS: Five of the children had residual phonological impairment. Only two children showed below average reading (decoding and comprehension), although five also showed below average spelling performance. Children with limitations in verbal memory, language production and metaphonology at the follow-up point were more likely to show delays in literacy. Language production skills at the follow-up point were most strongly correlated with literacy development. In terms of the early study, the strongest predictor for literacy development was performance on metaphonology tasks at the end of the early intervention study. The strongest predictor for ongoing speech impairment was phonological skill at the end of the early study. CONCLUSIONS: Results suggest that early phonological and metaphonological intervention can promote normalization of speech development and normal acquisition of literacy skills for at least some children with a history of severe phonological impairment. In the earlier study, the component structure of words (onsets, rhymes, codas) was emphasized through both metaphonological and non-linear phonological intervention. Risk for literacy and ongoing speech impairment can be reduced through early intervention that draws attention to the structure of words. PMID- 15832524 TI - Speech-production skills in children aged 3-7 years. AB - BACKGROUND: In recent years, clinicians have been using a psycholinguistic approach to the assessment and remediation of children's developmental speech disorders. This requires the comparison of a child's performance across a range of speech-production tasks. AIMS: To describe the profile of performance across different speech-production tasks in normal development and to discuss the application of such data to clinical findings. METHODS & PROCEDURES: Three speech production tasks, picture naming, word repetition and non-word repetition, were presented to 100 children with normal speech development, aged between 3 and 7 years of age. The speech-processing demands of the different tasks were considered using a developmental speech-processing model. Stimuli used in the three task paradigms were carefully matched so that children's performance across the tasks could be directly compared. OUTCOMES & RESULTS: Within the context of normal speech development, there were significant improvements in performance for all three tasks as children get older. There were also significant differences in performance across the three tasks, and the pattern of these relationships changed with age. Significant differences were found in the accuracy of production of words of increasing length. CONCLUSIONS: Profiles of speech production task performance are presented within the context of normal development. Comparison of performance across three different speech-production tasks might provide useful insight into the nature of a child's speech disorder. PMID- 15832525 TI - Semantic and inferencing abilities in children with communication disorders. AB - BACKGROUND: Semantic and inferencing abilities have not been fully examined in children with communication difficulties. AIMS: To investigate the inferential and semantic abilities of children with communication difficulties using newly designed tasks. METHODS & PROCEDURES: Children with different types of communication disorder were compared with each other and with three groups of typically developing children: those of the same chronological age and two groups of younger children. In total, 25 children aged 11 years with specific language impairment and 22 children, also 11 years of age, with primary pragmatic difficulties were recruited. Typically developing groups aged 11 (n=35; age match), and those aged 9 (n=40) and 7 (n=37; language similar) also participated as comparisons. OUTCOMES & RESULTS: For Semantic Choices, children with specific language impairment performed significantly more poorly than 9- and 11-year-olds, whilst the pragmatic difficulties group scored significantly lower than all the typically developing groups. Borderline differences between specific language impairment and pragmatic difficulties groups were found. For inferencing, children with communication impairments performed significantly below the 11-year old peers, but not poorer than 9- and 7-year-olds, suggesting that this skill is in line with language ability. Six children in the pragmatic difficulties group who met diagnosis for autism performed more poorly than the other two clinical groups on both tasks, but not statistically significantly so. CONCLUSIONS: Both tasks were more difficult for those with communication impairments compared with peers. Semantic but not inferencing abilities showed a non-significant trend for differences between the two clinical groups and children with pragmatic difficulties performed more poorly than all typically developing groups. The tasks may relate to each other in varying ways according to type of communication difficulty. PMID- 15832526 TI - Imitative production of regular past tense -ed by English-speaking children with specific language impairment. AB - BACKGROUND: Language intervention procedures often involve the speech-language pathologist highlighting or making more salient forms that are problematic for the child with a language impairment. According to limited processing accounts of specific language impairment (SLI), one way to increase the saliency of a form is to manipulate its sentence position. AIMS: The placement of verbs with past tense regular -ed, a morpheme documented to be significantly difficult to acquire for English-speaking children with SLI, was manipulated. Specifically, the accurate production of verbs with regular past tense -ed in the final position of sentences was evaluated in comparison with verbs with regular past tense -ed in the internal sentence position. METHODS & PROCEDURES: Ten English-speaking children with SLI (mean age 5;2) who were variable in the production of the regular past tense -ed morpheme participated in the study. A sentence imitation task that consisted of sentences with the past tense -ed in the sentence internal position, the sentence final position and foil sentences was presented. OUTCOMES & RESULTS: Overall, the data suggest that in an imitative context, errors of omission occurred only when regular past tense -ed occurred in the sentence internal position compared with the sentence final position. CONCLUSIONS: The results are consistent with the SA explanation for omitted tense marking in children with SLI and do not support the Extended Optional Infinitive Account. Assessment and intervention of past tense -ed may be improved by manipulating the sentence position of this morpheme. PMID- 15832527 TI - Challenges for undergraduate speech pathology students undertaking cross-cultural clinical placements. AB - BACKGROUND: Two Australian undergraduate speech pathology students completed a series of clinical placements working with people with complex communication needs in cross-cultural contexts. AIMS: To describe the challenges that the students faced and how best to prepare future students for such experiences. METHODS & PROCEDURES: The students completed the placements in Thailand, Nepal, Bangladesh, India and South Africa. They used personal journaling to record their experiences. The students used the journals as the basis for reflective discussion when considering the challenges they experienced in applying their knowledge and skills. OUTCOMES & RESULTS: The challenges were (1) to prepare adequately ahead of the placements; (2) to select appropriate models of service delivery; (3) to use existing service approaches, resources and infrastructure appropriately; (4) to access professional support; (5) to define their professional role; (6) to manage variable shared language proficiency; (7) to adapt personally; and (8) to work using an undergraduate level of knowledge, skills and experience. CONCLUSIONS: The students encountered challenges related to their clinical preparation and their capacity to adapt in both a professional and personal sense. Future students preparing to undertake such placements need to attain at least minimum clinical competencies before placements. They will be helped if they have some clinical experience in working with people with complex communication needs in cross-cultural contexts, information about the cultures they will visit, and if they set professional and personal learning goals for each clinical placement. This preparation will aid students in maximizing their learning experience. PMID- 15832528 TI - Patient autonomy: reflections from a developing country. PMID- 15832529 TI - Defending the use of EBM and shared decision making. PMID- 15832530 TI - Case report: insect bite reveals botfly myiasis in an older woman. PMID- 15832531 TI - Prenatal care: examining the evidence for an evolving paradigm. PMID- 15832532 TI - Antibiotics for recurrent urinary tract infections. PMID- 15832533 TI - Effects of discontinuing epidurals in late labor. PMID- 15832534 TI - Evidence-based prenatal care: Part I. General prenatal care and counseling issues. AB - Effective prenatal care should integrate the best available evidence into a model of shared decision making. Pregnant women should be counseled about the risks of smoking and alcohol and drug use. Structured educational programs to promote breastfeeding are effective. Routine fetal heart auscultation, urinalysis, and assessment of maternal weight, blood pressure, and fundal height generally are recommended, although the evidence for these interventions is variable. Women should be offered ABO and Rh blood typing and screening for anemia during the first prenatal visit. Genetic counseling and testing should be offered to couples with a family history of genetic disorders, a previously affected fetus or child, or a history of recurrent miscarriage. All women should be offered prenatal serum marker screening for neural tube defects and aneuploidy. Women at increased risk for aneuploidy should be offered amniocentesis or chorionic villus sampling. Counseling about the limitations and risks of these tests, as well as their psychologic implications, is necessary. Folic acid supplementation beginning in the preconception period reduces the incidence of neural tube defects. There is limited evidence that routine use of other dietary supplements may improve outcomes for the mother and infant. PMID- 15832535 TI - Information from your family doctor. Pregnancy: keeping yourself and your baby healthy. PMID- 15832536 TI - Evaluation of the patient with muscle weakness. AB - Muscle weakness is a common complaint among patients presenting to family physicians. Diagnosis begins with a patient history distinguishing weakness from fatigue or asthenia, separate conditions with different etiologies that can coexist with, or be confused for, weakness. The pattern and severity of weakness, associated symptoms, medication use, and family history help the physician determine whether the cause of a patient's weakness is infectious, neurologic, endocrine, inflammatory, rheumatologic, genetic, metabolic, electrolyte-induced, or drug-induced. In the physical examination, the physician should objectively document the patient's loss of strength, conduct a neurologic survey, and search for patterns of weakness and extramuscular involvement. If a specific cause of weakness is suspected, the appropriate laboratory or radiologic studies should be performed. Otherwise, electromyography is indicated to confirm the presence of a myopathy or to evaluate for a neuropathy or a disease of the neuromuscular junction. If the diagnosis remains unclear, the examiner should pursue a tiered progression of laboratory studies. Physicians should begin with blood chemistries and a thyroid-stimulating hormone assay to evaluate for electrolyte and endocrine causes, then progress to creatine kinase level, erythrocyte sedimentation rate, and antinuclear antibody assays to evaluate for rheumatologic, inflammatory, genetic, and metabolic causes. Finally, many myopathies require a biopsy for diagnosis. Pathologic evaluation of the muscle tissue specimen focuses on histologic, histochemical, electron microscopic, biochemical, and genetic analyses; advances in technique have made a definitive diagnosis possible for many myopathies. PMID- 15832537 TI - Diagnosis of gastrointestinal bleeding in adults. AB - The clinical evaluation of gastrointestinal bleeding depends on the hemodynamic status of the patient and the suspected source of the bleeding. Patients presenting with upper gastrointestinal or massive lower gastrointestinal bleeding, postural hypotension, or hemodynamic instability require inpatient stabilization and evaluation. The diagnostic tool of choice for all cases of upper gastrointestinal bleeding is esophagogastroduodenoscopy; for acute lower gastrointestinal bleeding, it is colonoscopy, or arteriography if the bleeding is too brisk. When bleeding cannot be identified and controlled, intraoperative enteroscopy or arteriography may help localize the bleeding source, facilitating segmental resection of the bowel. If no upper gastrointestinal or large bowel source of bleeding is identified, the small bowel can be investigated using a barium-contrast upper gastrointestinal series with small bowel follow-through, enteroclysis, push enteroscopy, technetium-99m-tagged red blood cell scan, arteriography, or a Meckel's scan. These tests may be used alone or in combination. PMID- 15832538 TI - Methadone treatment for pain states. AB - Methadone is a synthetic opioid with potent analgesic effects. Although it is associated commonly with the treatment of opioid addiction, it may be prescribed by licensed family physicians for analgesia. Methadone's unique pharmacokinetics and pharmacodynamics make it a valuable option in the management of cancer pain and other chronic pain, including neuropathic pain states. It may be an appropriate replacement for opioids when side effects have limited further dosage escalation. Metabolism of and response to methadone varies with each patient. Transition to methadone and dosage titration should be completed slowly and with frequent monitoring. Conversion should be based on the current daily oral morphine equivalent dosage. After starting methadone therapy or increasing the dosage, systemic toxicity may not become apparent for several days. Some medications alter the absorption or metabolism of methadone, and their concurrent use may require dosing adjustments. Methadone is less expensive than other sustained-release opioid formulations. PMID- 15832539 TI - Screening for thyroid disease. PMID- 15832540 TI - St. John's wort for depression. PMID- 15832541 TI - Pruritic rash during pregnancy. PMID- 15832542 TI - Withdrawal of older anticonvulsants for management of status epilepticus: implications for resource-poor countries. PMID- 15832543 TI - Skeletal maturation in cerebral palsy. PMID- 15832544 TI - Neurobehavioural profile of low-birthweight infants with cystic periventricular leukomalacia. AB - Twenty-three low-birthweight infants (17 males, six females) diagnosed with cystic periventricular leukomalacia (PVL; median gestational age 30 wks, postmenstrual age range 25 to 36 wks; median birthweight 1365 g, range 680 to 2010 g) were evaluated and compared with 209 comparison infants (117 males, 92 females; median gestational age 33 wks, postmenstrual age range 25 to 39 wks; birthweight 1771 g, range 670 to 2460 g). There were three assessment times: 36 to 38 weeks (preterm), 40 to 42 weeks (term), and 44 to 46 weeks (post-term); the Neonatal Behavioral Assessment Scale (NBAS) was used for assessment. Outcome at 2 years was assessed on the basis of a neurological examination, magnetic resonance imaging, computerized tomography, electroencephalography, and the Mental and Psychomotor Development Indices of the Bayley Scales of Infant Development. All infants in the PVL group were found to have evidence of cerebral palsy, whereas all infants in the comparison group were normally developing. Infants with PVL performed more poorly on all elements of the NBAS examination at all three assessment times compared with the comparison group. They demonstrated poorer motor control, less responsiveness to environmental stimuli, less regulatory capacity, and more abnormal reflexes compared with the comparison group. These results suggest that LBW infants with PVL show dysfunction and/or disorganization in their neurobehavioural systems in the neonatal period. Assessment of neonatal neurobehavioural characteristics using the NBAS may assist clinicians in identifying LBW infants with PVL, and in formulating plans for the developmental care of these infants. PMID- 15832545 TI - Altered skeletal maturation in moderate to severe cerebral palsy. AB - Abnormalities of growth and development are prevalent in children with cerebral palsy (CP). The purpose of this study was to assess skeletal maturation ('bone age') in this population and to identify those factors related to alterations in this aspect of development. The study group was a convenience sample of 80 participants (47 males, 33 females; age range 2 y 6 mo to 21 y 1 mo; mean age 10 y 10 mo, SD 4 y 2 mo). All had moderate to severe spastic or mixed spastic CP at Gross Motor Function Classification System level III (n=8, 10%); level IV (n=21, 26%); or level V (n=51, 64%). Skeletal maturation was assessed by the detailed Fels method of scoring hand-wrist radiographs. Each evaluation included clinical and anthropometric assessments, Tanner staging, bone density measurements, and the Children's Health Status Questionnaire. Follow-up evaluations were obtained for 41 participants, providing a total of 143 evaluations. Median interval between first and last evaluations was 24.7 months (range 11.9 to 45mo). For the study group as a whole there was no significant difference between a child's skeletal age and chronological age. However, there was a high prevalence of individual participants in whom skeletal age was advanced (7%) or delayed (10%), relative to chronological age, by more than 2 years. In multivariant analyses it was found that diminished linear growth (height), low lumbar-spine bone density, and low body fat as measured by triceps skinfolds were all independently associated with delays in skeletal maturation. Multiple aspects of skeletal growth and development, including skeletal maturation, are frequently altered in children with moderate to severe CP. PMID- 15832546 TI - Psychological disturbance and sleep disorders in children with neurofibromatosis type 1. AB - The objective of this study was to explore the behaviour, including sleep patterns, of children with neurofibromatosis type 1 (NF1). For this purpose we designed a cross-sectional descriptive survey conducted by postal enquiry with telephone follow-up. Simonds and Parraga's sleep questionnaire and the Strength and Difficulties Questionnaire were used to screen sleep patterns and behaviours respectively. Information was obtained for 64 children (39 male, mean age 10 y 7 mo, SD 4 y 1 mo, range 3 to 18 y; 57% response rate). There were increased numbers of children with NF1 achieving scores in the borderline and abnormal range for ratings of peer problems (p < 0.001), hyperactivity (p < 0.001), emotional symptoms (p < 0.001), and conduct disorder (p < 0.05). Total difficulties score was also high (p < 0.001) in the NF1 group. Only one group of sleep problems, parasomnias (in particular sleepwalking and sleep terrors), had a higher occurrence (p < or = 0.05) in the NF1 group than in the general population. Within the NF1 group, conduct problems (p < or = 0.05), hyperactivity (p < or = 0.01), emotional problems (p < or = 0.01), and the total difficulties score (p < or = 0.01) were all significantly higher in children with frequent sleep disturbance. We conclude that NF1 is associated with sleep and behavioural problems in a high proportion of children. Psychiatric conditions, e.g. attention deficit-hyperactivity disorder, may be underdiagnosed on a large scale in children with NF1, and the use of simple screening tools in clinical settings may prove beneficial. PMID- 15832547 TI - Refractive errors in infancy predict reduced performance on the movement assessment battery for children at 3 1/2 and 5 1/2 years. AB - We have previously reported that significant hyperopia at 9 months predicts mild deficits on visuocognitive and visuomotor measures between 2 years and 5 years 6 months. Here we compare the motor skills of children who had been hyperopic in infancy (hyperopic group) with those who had been emmetropic (control group), using the Movement Assessment Battery for Children (Movement ABC). Children were tested at 3 years 6 months (hyperopic group: 47 males, 63 females, mean age 3 y 7 mo, SD 1.6 mo; control group: 61 males, 70 females, mean age 3 y 7 mo, SD 1.2 mo) and at 5 years 6 months (hyperopic group: 43 males, 56 females, mean age 5 y 4 mo, SD 1.7 mo; control group: 51 males, 62 females, mean age 5 y 3 mo, SD 1.6 mo). The hyperopic group performed significantly worse at both ages, overall and on at least one test from each category of motor skill (manual dexterity, balance, and ball skills). Distributions of scores showed that these differences were not due to poor performance by a minority but to a widespread mild deficit in the hyperopic group. This study also provides the first normative data on the Movement ABC for children below 4 years of age, and shows that it provides a useful measure of motor development at this young age. PMID- 15832548 TI - A proposed classification for subtypes of arterial ischaemic stroke in children. AB - The aim of this study was to propose a classification system for childhood arterial ischaemic stroke (AIS). Subtypes from the Trial of Org 10172 in Acute Stroke Therapy (TOAST) classification, previously shown to be applicable to children, were retained in the proposed Paediatric Stroke Classification (PSC). Additional important paediatric AIS aetiologies were identified from a literature review. Preliminary validation was performed by three raters who categorized clinical vignettes from 135 patients (66 male; median age 6.3 y, range 0.1 to 16 y). Eight aetiological subtypes were identified and defined, as follows: (1) sickle cell disease; (2) cardioembolic; (3) moyamoya syndrome; (4) cervical arterial dissection; (5) steno-occlusive cerebral arteriopathy; (6) other determined aetiology; (7) multiple probable/possible aetiologies; and (8) undetermined aetiology. There was very good agreement between the raters about categorization of the vignettes. Causes of disagreement were identified and final categories and definitions were modified accordingly. We conclude that the PSC enables the categorization of children with AIS into aetiological subtypes relevant to this age group. This will be useful in multicentre studies of natural history and treatment but will require further independent validation. PMID- 15832549 TI - Association between intellectual functioning and age in children and young adults with Duchenne muscular dystrophy: further results from a meta-analysis. AB - This study used meta-analytical techniques to explore the association between intelligence and age in children with Duchenne muscular dystrophy (DMD). The sample comprised 1224 children and young adults with DMD (mean age 12 y 3 mo, SD 4 y; range 2 y to 27 y). Standardized measures including the Wechsler Intelligence Scales (WIS) and the Stanford-Binet Intelligence Scales were used to estimate intelligence. No age-related difference was noted for Full-scale and Performance intelligence quotients (IQ). However, Verbal IQ increased significantly with age. Age-related increases were noted for the WIS Information, Similarities, Arithmetic, Comprehension, Digit Span, Picture Arrangement, Block Design, and Coding subscales. These results support the notion that younger children with DMD have deficits in verbal reasoning and verbal processing. Older children with DMD, particularly those of 14 years and older, were less likely to present with these problems. Implications of these findings and possible future research directions are discussed. PMID- 15832550 TI - Effects of constraint-induced movement therapy in young children with hemiplegic cerebral palsy: an adapted model. AB - The aim of this study was to evaluate the effects of a modified version of constraint-induced (CI) movement therapy on bimanual hand-use in children with hemiplegic cerebral palsy (CP; age range 18 mo to 4 y) and to make a comparison with conventional paediatric treatment. Twenty-one children (13 females, eight males) completed the CI therapy programme and 20 children (12 males, eight females) served as a control group. Children in the CI therapy group were expected to wear a restraint glove for 2 hours each day over a period of 2 months. The training was based on principles of motor learning used in play and in motivational settings. To evaluate the effect of treatment, the Assisting Hand Assessment (AHA) was used. Assessments took place on three occasions: at onset, after 2 months, and 6 months after the first assessment. A significant interaction was found between group and AHA measure (ANOVA, F(2,74) = 5.64, p = 0.005). The children who received CI therapy improved their ability to use their hemiplegic hand significantly more than the children in the control group after 2 months, i.e. after treatment. Effect size was high after treatment and remained medium at 6 months. As the treatment was tailored to each child's capacity and interests, little frustration was experienced by the children. PMID- 15832551 TI - Visual development in preterm infants. PMID- 15832552 TI - Subjective quality of life in children with intellectual impairment--how can it be assessed? PMID- 15832553 TI - Breathing patterns of athletes with cerebral palsy. PMID- 15832554 TI - Is a vaccine needed against Candida albicans? AB - The development of a useful Candida vaccine is a distinct possibility despite the fact that individuals with a lifetime of commensal sensitization do not develop sterile immunity to the organism. An effective Candida vaccine would be invaluable in preventing hematogenously disseminated candidiasis, as well as mucocutaneous disease. This review is a discussion of our current understanding of the interplay between commensal and pathogenic forms of Candida albicans and approaches toward active and passive immunoprevention against candidiasis. PMID- 15832555 TI - Geographic information system analysis of blastomycosis in northern Wisconsin, USA: waterways and soil. AB - Geographic information systems (GIS) are powerful tools for investigating the ecogeography of environmentally acquired infections. GIS technology was used to geocode and map cases of blastomycosis, by household, of human and dog residents of Vilas County, Wisconsin, USA. Human case households (n = 136) were from a comprehensive street address registry 1979--2001; human controls were 200 random number selected households from 2001 county tax records. Dog cases (n = 116) were from a consecutive street address registry from a private veterinarian practice, Eagle River; dog controls were 200 random-number selected addresses from the 2001 total practice registry. Where geocoding was not available from existing maps, home sites were visited and geocoded using a handheld global positioning system (n = 61). Waterway characteristics were obtained from the Wisconsin Department of Natural Resources GNIS database, soil type data from the USDA. Data were analysed with chi-square and Mann-Whitney tests; cluster analysis with CrimeStat II software. A disproportionate number of human and dog cases were associated with waterways of the western Eagle River area. Human and dog cases were more commonly near waterways < 500 m elevation (more nutrient rich) (P < or =0.001 for both) and were associated with sands (prone to drought) (P < or = 0.01 for both). When the nearest waterway was a lake, case addresses were more commonly near lakes with <7.6 m maximum depth (more lake mixing) (P <0.05, humans and dogs) but mean depth, type and size of lakes did not differ from controls. Further studies are needed to clarify the nature of these associations. PMID- 15832556 TI - Possible differences in pathogenicity between cane toad-, frog- and platypus derived isolates of Mucor amphibiorum, and a platypus-derived isolate of Mucor circinelloides. AB - Platypuses (Ornithorhynchus anatinus) in the north of the island state of Tasmania, Australia, suffer from a serious disease called ulcerative mycosis, which is responsible for high morbidity and, presumably, mortality rates in areas where it occurs. The disease is caused by the dimorphic fungus Mucor amphibiorum, which is also found in Queensland, New South Wales and Victoria. However, it does not cause disease in platypuses in those states. It has been previously reported that a closely related fungus, Mucor circinelloides, may also be capable of causing this disease. This paper describes pathogenicity trials involving cane toads (Bufo marinus) as the experimental model. The toads were infected with either Tasmanian, platypus-derived M. amphibiorum, West Australian, frog-derived M. amphibiorum, Queensland cane-toad-derived M. amphibiorum or Tasmanian platypus derived M. circinelloides. The Tasmanian isolates of M. amphibiorum were more likely to cause a serious, long-term infection than were Queensland or West Australian isolates, and (+) mating types caused a more serious infection than the (-) mating type. The isolate of M. circinelloides was incapable of infecting the toads, lending further weight to the theory that it represents an environmental contaminant. The results suggest that an endemic strain of M. amphibiorum has mutated and become pathogenic to platypuses. Alternatively, a pathogenic strain of M. amphibiorum may have been introduced into Tasmania, where it is infecting a naive population. PMID- 15832557 TI - Assessing in vitro combinations of antifungal drugs against yeasts and filamentous fungi: comparison of different drug interaction models. AB - Non-parametric and parametric approaches of two competing zero-interaction theories--the Loewe additivity and the Bliss independence - were evaluated for analyzing the in vitro interactions of various antifungal drugs. Fifty-one data sets, derived from three drug combinations, tested in triplicate against 17 clinical yeast and mold isolates with a two-dimensional checkerboard microdilution technique, were selected to span from strong synergy to strong antagonism. These were analyzed with the standard FIC index model and modern concentration-effect response surface models: the fully parametric model developed by Greco et al. and the 3-D analysis developed by Prichard et al. The FIC index model is subjective, sensitive to experimental errors and resulted in approximated results and variable conclusions depending on the MIC endpoints determined and interpretation endpoints used. By using the MIC-2 endpoint (lowest drug concentration showing 50% of growth) for calculating the FIC indices, problems due to trailing phenomena were reduced and weak interactions could be detected; higher levels of reproducibility and agreement with the other models were achieved using the MIC-0 and MIC-1 (lowest drug concentration showing 10 and 25% of growth, respectively). High reproducibility was achieved in interpreting the FIC indices when the cutoffs of 0.25 and 4 (for single experiments) and the cutoff of 1 (for replicates) were used for defining the limits of additivity/indifference. Although the fully parametric Greco model did not describe precisely the entire response surface of all antifungal drug interactions, it was able to differentiate synergistic from non-synergistic interactions with a non-unit, reproducible, concentration-independent interaction parameter, including its uncertainty, without requiring replication. The Bliss independence based models resulted in mosaics of synergistic and antagonistic combinations, raising questions about the concentration-dependent nature of antifungal drug interaction. The sum of all statistically significant interactions were used as a summary interaction parameter for the entire response surface, concluding synergy or antagonism when it was positive or negative, respectively. The cutoffs of 100% and 200% were used to distinguish weak and moderate interactions, respectively in 12-16 x 8-12 checkerboard formats. Semi parametric approaches need particular care as experimental errors are not eliminated from the entire response surface. PMID- 15832558 TI - alpha-galf I -->6-alpha-mannopyranoside side chains in Paracoccidioides brasiliensis cell wall are shared by members of the Onygenales, but not by galactomannans of other fungal genera. AB - The water-soluble polysaccharide fraction of the cell wall alkali extract (F1SS) from the mycelial phase of the dimorphic fungus Paracoccidioides brasiliensis is compared with F1SS polysaccharides obtained from the Onygenalean mycelial fungi Ascocalvatia alveolata, Onygena equina and Aphanoascus terreus. These polymers were exclusively composed of mannose and galactose. Data from methylation and NMR analyses reveal that F1SS polysaccharides from the four fungi contain the same residues although in different proportions: [-->2,6)-alpha-D-Manp-(1 -->]; [2) alpha-D-Manp-(1 -->]; [ -->6)-alpha-D-Manp-(1 -->]; and [alpha-D-Galf-(1 -->]. In P. brasiliensis, the repeating unit of the polysaccharide consists of a backbone of [(1 -->6)-alpha-D-Manp] substituted at the 0-2 position by the disaccharide [alpha-D-Galf-(1 -->6)-alpha-D-Manp-(1 -->], while the remaining 0-2 positions are substituted by single residues of mannose or short chains of (1 -->2) mannose. The other species had a lower proportion of galactofuranose-containing side chains and higher proportion of mannose-containing side chains. The similarities found among the F1SS polysaccharides from P. brasiliensis and the Onygenalean A. alveolata, A. terreus and O. equina, reveal the close relatedness of all these fungi, show differences with polysaccharides from other fungal genera and agree with the molecular evidence provided in the scientific literature for the placement of P. brasiliensis within the Onygenales. PMID- 15832559 TI - Chrysosporium chiropterorum sp. nov., isolated in France, resembling Chrysosporium state of Ajellomyces capsulatus (Histoplasma capsulatum). AB - A fungus isolated in France from the fur of a bat, which produces characterized large tuberculate conidia (aleurioconidia) similar to those produced by the mycelial form of Histoplasma capsulatum (Ajellomyces capsulatus) is described. Colonies are white at first, but then become rosy buff from the centre outwards. Sectoring, resulting in the appearance of patches or areas of dark green mycelium, occurs spontaneously. Single-celled conidia are formed on undifferentiated hyphae, and may be sessile, or borne laterally on short stalks or producing in an intercalary position as it is the case in the genus Chrysosporium. This fungus is clearly distinguishable from any described species and is described as Chrysosporium chiropterorum sp. nov. C. chiropterorum, like H. capsulatum, produces gelatinase, and is non-keratinolytic but strongly ureolytic. Both species are associated with bat dwellings. C. chiropterorum differs from H. capsulatum by faster growth, pink or green colonies, and failure to produce microconidia as well as lack of conversion to a yeast phase in vitro at 37 degrees C. PMID- 15832560 TI - In vitro activity of caspofungin compared to amphotericin B, fluconazole, and itraconazole against Candida strains isolated in a Turkish University Hospital. AB - We investigated the in vitro activity of caspofungin compared to amphotericin B, fluconazole, and itraconazole against clinical strains of Candida spp. (n =239). Antifungal susceptibility tests were done in accordance with NCCLS M27-A2 microdilution method and the results were read after 24 and 48 h. In general, 24 h MIC readings were similar to those at 48 h for most isolates and all antifungal agents. Caspofungin was active against all species tested. Caspofungin MICs of Candida parapsilosis were slightly higher than those for other Candida spp. Caspofungin MIC (microg/ml) ranges at 24 h for C. albicans, C. glabrata, C tropicalis, C. parapsilosis, C kefyr, C krusei, C. lusitaniae, C. norvegensis, C. guilliermondii and C. lipolytica were 0.06-2, 0.125-2, 0.125-2, 1-4, 0.125-2, 1 2, 0.5-2, 0.5-1, 0.5-2 and 1-2, respectively. Eagle (paradoxical) effect was observed in 31 and 8% of the isolates at highest concentrations of caspofungin and itraconazole, respectively. The activity of caspofungin against fluconazole- and/or itraconazole-resistant isolates was similar to that detected for the susceptible ones. We conclude that caspofungin appears as a promising antifungal agent with enhanced activity against Candida, including the azole-resistant strains. PMID- 15832561 TI - In vitro activities of posaconazole, ravuconazole, terbinafine, itraconazole and fluconazole against dermatophyte, yeast and non-dermatophyte species. AB - The in vitro activities of two new triazole antifungal agents with broad-spectrum antifungal activity, posaconazole and ravuconazole, were compared with those of three well-established antifungal agents, terbinafine, itraconazole and fluconazole, against 184 clinical isolates. These included 129 dermatophyte isolates (twelve species), 25 yeast isolates (five species) and 28 non dermatophyte isolates (nine species). In vitro testing was conducted using microdilution plates with RPMI 1640 and National Committee for Clinical Laboratory Standards (NCCLS) guidelines (M27-38P) were followed, except for the preparation of the dermatophyte inoculum. Both posaconazole and ravuconazole showed similar broad-spectrum activity against dermatophyte, yeast and non dermatophyte species. Mean inhibitory concentrations (MIC) at which 90% [MIC90] of the isolates were inhibited by posaconazole and ravuconazole were 0.25 and 0.5 microg/ml for dermatophytes, 0.5 and 0.25 microg/ml for yeasts, and >4 and 8 microg/ml for non-dermatophytes. The MIC ranges against Trichophyton (six species), Microsporum (five species) and Epidermophyton flocossum were: posaconazole (0.007-1.0/0.007-0.25/0.007-1.0 microg/ml), ravuconazole (0.015 8.0/0.015-1.0/0.015-1.0 microg/ml), itraconazole (0.015- >8.0/0.015-0.5/ 0.015 8.0 microg/ml), fluconazole (0.125- >64.0/4.0 >64.0/0.5-64.0 microg/ml) and terbinafine (0.003 >2.0/0.007-2.0/0.007 >2.0 microg/ml). Overall ranking of the antifungal activity of the five antifungal agents was: terbinafine > posaconazole > ravuconazole > itraconazole > fluconazole, for dermatophytes; ravuconazole > posaconazole > itraconazole > fluconazole > terbinafine, against yeasts; and posaconazole > ravuconazole > terbinafine > itraconazole > fluconazole, for non dermatophytes. PMID- 15832562 TI - Multifocal sporotrichosis with meningeal involvement in a patient with AIDS. AB - This report describes a 29-year-old man with AIDS and disseminated lymphocutaneous sporotrichosis diagnosis presenting a poor therapeutic adhesion to itraconazole therapy that later evolved to neurological impairment and death. Necropsy showed granulomatous reaction and yeast-like forms similar to Sporothrix schenckii in meninges, lymph nodes, marrow bone, skin, testicles, epididymides and pancreas. Meningeal sporotrichosis cases associated to AIDS are reviewed in brief. PMID- 15832563 TI - Tale of a table: ins and outs of perception. PMID- 15832564 TI - Variations in backward masking with different masking stimuli: I. Local interaction versus attentional switch. AB - The types of stimuli used as targets and masks considerably change the masking functions in a way that requires us to abandon any single mechanism of masking as the sole explanation of backward masking. In the first of two reports in which the problem of the mask-dependence of masking is addressed, we explore the role of the relative spatial positioning of targets and masks in order to differentiate between local interaction and attentional models. If single letters were masked by double-letter masks then the relative spatial arrangement of the letters, which was changed in order to vary the involvement of metacontrast-like processes, had an effect at shorter SOAs, but not at longer SOAs where strong masking still persisted. This poses difficulties for proposing local contour interaction as the main mechanism of masking. Similarly, crowding effects alone cannot explain the results. Backward masking also involves attention being directed to working-memory processing of the succeeding object while abandoning the preceding object. PMID- 15832565 TI - Variations in backward masking with different masking stimuli: II. The effects of spatially quantised masks in the light of local contour interaction, interchannel inhibition, perceptual retouch, and substitution theories. AB - In part I we showed that with spatially non-overlapping targets and masks both local metacontrast-like interactions and attentional processes are involved in backward masking. In this second part we extend the strategy of varying the contents of masks to pattern masking where targets and masks overlap in space, in order to compare different masking theories. Images of human faces were backward masked by three types of spatially quantised masks (the same faces as targets, faces different from targets, and Gaussian noise with power spectra typical for faces). Configural characteristics, rather than the spectral content of the mask, predicted the extent of masking at relatively long stimulus onset asynchronies (SOAs). This poses difficulties for the theory of transient-on-sustained inhibition as the principal mechanism of masking and also for local contour interaction being a decisive factor in pattern masking. The scale of quantisation had no effect on the masking capacity of noise masks and a strong effect on the capacity of different-face masks. Also, the decrease of configural masking with an increase in the coarseness of the quantisation of the mask highlights ambiguities inherent in the re-entrance-based substitution theory of masking. Different masking theories cannot solve the problems of masking separately. They should be combined in order to create a complex, yet comprehensible mode of interaction for the different mechanisms involved in visual backward masking. PMID- 15832566 TI - Adaptation to spiral motion in crowding condition. AB - When a single, moving stimulus is presented in the peripheral visual field, its direction of motion can be easily distinguished, but when the same stimulus is flanked by other similar moving stimuli, observers are unable to report its direction of motion. In this condition, known as 'crowding', specific features of visual stimuli do not access conscious perception. The aim of this study was to investigate whether adaptation to spiral motion is preserved in crowding conditions. Logarithmic spirals were used as adapting stimuli. A rotating spiral stimulus (target spiral) was presented, flanked by spirals of the same type, and observers were adapted to its motion. The observers' task was to report the rotational direction of a directionally ambiguous motion (test stimulus) presented afterwards. The directionally ambiguous motion consisted of a pair of spirals flickering in counterphase, which were mirror images of the target spiral. Although observers were not aware of the rotational direction of the target and identified it at chance levels, the direction of rotation reported by the observers during the test phase (motion aftereffect) was contrarotational to the direction of the adapting spiral. Since all contours of the adapting and test stimuli were 90 degrees apart, local motion detectors tuned to the directions of the mirror-image spiral should fail to respond, and therefore not adapt to the adapting spiral. Thus, any motion aftereffect observed should be attributed to adaptation of global motion detectors (ie rotation detectors). Hence, activation of rotation-selective cells is not necessarily correlated with conscious perception. PMID- 15832567 TI - Configural processing and perceptions of head tilt. AB - Configural processing is important for face recognition, but its role in other types of face processing is unclear. In the present study, participants made judgments of head tilt for faces in which the vertical position of the internal facial region was varied. We found a highly reliable relationship between inner face position and perceived head tilt. We also found that changes in inner-face position affected the perceived dimensions of an individual unchanged facial feature: compared to control faces, nearly two-thirds of faces in which the features had been moved down were judged to have a longer nose. This finding suggests an early integration of configural and featural processing to create a stable holistic percept of the face. The demonstration of holistic processing at a basic perceptual level (as opposed to during face recognition) is important as it constrains possible models of the relationships between featural and configural processing. PMID- 15832568 TI - Spatial balance of color triads in the abstract art of Piet Mondrian. AB - We examined the interactive contribution of the color and size of the three areas occupied by the primary colors red, yellow, and blue in adaptations of abstract compositions by Mondrian to the perceived weight of the areas and the location of the balance centers of the compositions. Thirty-six art stimuli were created by experimentally changing the colors in the three areas of six original works so that the resulting five variations and the original constituted the six possible spatial arrangements of the three colors in the three locations. In experiment 1, design-trained and untrained participants determined the location of the balance center of each composition seen on a computer screen and rated the apparent weight or heaviness of each color area. In experiment 2, untrained participants determined the location of the balance centers of the compositions when projected to their actual size. It was found that, for both trained and untrained participants, the perceived weight of a color, especially red and yellow, varied as a function of the size of the area it occupied. Furthermore, participants in both experiments perceived shifts in the locations of the balance centers between the originals and their altered versions. Only the trained participants, however, perceived significant shifts in balance centers among the five variations of the compositions, demonstrating their superior sensitivity to the contribution of color to balance structure. Taken together, the findings demonstrate the existence of a color-area-weight relationship among color triads in abstract displays and the influence of this relationship on color balance in abstract compositions. PMID- 15832569 TI - The influence of restricted viewing conditions on egocentric distance perception: implications for real and virtual indoor environments. AB - We carried out three experiments to examine the influence of field of view and binocular viewing restrictions on absolute distance perception in real-world indoor environments. Few of the classical visual cues provide direct information for accurate absolute distance judgments to points in the environment beyond about 2 m from the viewer. Nevertheless, in previous work it has been found that visually directed walking tasks reveal accurate distance estimations in full-cue real-world environments to distances up to 20 m. In contrast, the same tasks in virtual environments produced with head-mounted displays (HMDs) show large compression of distance. Field of view and binocular viewing are common limitations in research with HMDs, and have been rarely studied under full pictorial-cue conditions in the context of distance perception in the real-world. Experiment 1 showed that the view of one's body and feet on the floor was not necessary for accurate distance perception. In experiment 2 we manipulated the horizontal and the vertical field of view along with head rotation and found that a restricted field of view did not affect the accuracy of distance estimations when head movement was allowed. Experiment 3 showed that performance with monocular viewing was equal to that with binocular viewing. These results have implications for the information needed to scale egocentric distance in the real world and reduce the support for the hypothesis that a limited field of view or imperfections in binocular image presentation are the cause of the underestimation seen with HMDs. PMID- 15832570 TI - Speechreading skill and visual movement sensitivity are related in deaf speechreaders. AB - Individual speechreading abilities have been linked with a range of cognitive and language-processing factors. The role of specifically visual abilities in relation to the processing of visible speech is less studied. Here we report that the detection of coherent visible motion in random-dot kinematogram displays is related to speechreading skill in deaf, but not in hearing, speechreaders. A control task requiring the detection of visual form showed no such relationship. Additionally, people born deaf were better speechreaders than hearing people on a new test of silent speechreading. PMID- 15832571 TI - Does selective attention influence the octave illusion? AB - The octave illusion occurs when each ear receives a sequence of tones alternating by one octave, but with different frequencies in each ear. Most listeners report a high pitch in one ear alternating with a low pitch in the opposite ear. Deutsch and Roll proposed an influential suppression model of the illusion in which the pitch is determined by ear dominance, while the location of this pitch is determined by high-frequency dominance. Deutsch later suggested that this unusual division between 'what' and 'where' mechanisms is facilitated by sequential interactions within the eliciting sequence. A recent study has raised doubts about the suppression model and the role of sequential interactions in the illusion (Chambers et al, 2002 Journal of Experimental Psychology: Human Perception and Performance 28 1288-1302). Here, we examined whether this previous null effect of sequential interactions may have arisen because of uncontrolled influences of selective attention. The results reveal no evidence of a link between selective attention and sequential interactions, thus consolidating doubts about the validity of the suppression model. PMID- 15832572 TI - Modification of magnitude estimations in thermotactile perception during self generated and externally generated movements. AB - Fourteen participants felt a 'cold' stimulus move across a fingertip. When movement was self-controlled, the stimulus was reported as feeling less 'cold' than when movement was externally generated. PMID- 15832573 TI - Last but not least. AB - When two three-letter words are flashed up in sequence, observers cannot tell whether the top halves of the words are the same or different. It follows that words, like faces, are processed holistically, not as a set of separate features. PMID- 15832574 TI - Dynamic accuracy survey of the new "single plane--single frame 3-D calibration" technique for use in biomedical applications. AB - This study tested the accuracy of a new 3-D calibration technique under dynamic situations. The technique was firstly introduced in 1998 for biomechanical human tests and calibrates 3-D volumes in an easy way. It revealed superior in static tests to others. In order to disclose dynamic accuracy two different tests were performed. With this technique it does not matter whether redundant information from multiple camera views is available or not. The mean error for distances measured at 0.018% for redundant information and at 0.012% for the non-redundant test in contrast to other procedures found in literature, which attain values of 0.09% and 0.04% respectively. The maximum error ranged there between 5.5% and 17.9%, whereas the presented data reached values of 0.33% and 0.48%. The more important angle error was at maximum 0.055% (9 times less than the most accurate in literature) and nearly zero for the mean error value. The level of noise was the same in the test with redundancy and 7.4 times lower in the present study than other commercial available systems for non-redundant video information. The new procedure revealed as a stable and very accurate 3-D reconstruction technique for a variety of application not limited to biomedical applications. PMID- 15832575 TI - [Assessment of precision and accuracy of digital surface photogrammetry with the DSP 400 system]. AB - The objective of the present study was to evaluate the precision and accuracy of facial anthropometric measurements obtained through digital 3-D surface photogrammetry with the DSP 400 system in comparison to traditional 2-D photogrammetry. Fifty plaster casts of cleft infants were imaged and 21 standard anthropometric measurements were obtained. For precision assessment the measurements were performed twice in a subsample. Accuracy was determined by comparison of direct measurements and indirect 2-D and 3-D image measurements. Precision of digital surface photogrammetry was almost as good as direct anthropometry and clearly better than 2-D photogrammetry. Measurements derived from 3-D images showed better congruence to direct measurements than from 2-D photos. Digital surface photogrammetry with the DSP 400 system is sufficiently precise and accurate for craniofacial anthropometric examinations. PMID- 15832576 TI - Comparison of X-ray fluorescence and optical fluorescence measured behind scattering layers as a basis for X-ray fluorescence endoscopy. AB - Recent developments in the technology of capillary-fiber optics suitable for X rays in the range of approximately 4-10keV point to the possible realization of endoscopes applicable in X-ray fluorescence analysis. A general problem is the determination of scattering and absorption processes with consideration to tissue optics, X-ray scattering and X-ray absorption in a diagnostic partial volume. Therefore comparative investigations were performed in order to answer these questions. Zinc-oxide nanoparticles configured as single particles and ZnO clusters provided the fluorescence source in cell layers. An artificial scattering material was employed, which closely approximated the tissue optical conditions and the X-ray optical application conditions in possible diagnostic situations. As a result imaging of spatially resolved X-ray contrasts was better than adequate optical fluorescence imaging by approximately one magnitude. Hence a very important precondition for realizing X-ray fluorescence endoscopy is fulfilled. PMID- 15832577 TI - [Micro-CT analysis of cancellous bone fragments from the distal radius fracture zone in osteoporosis]. AB - The incidence and prevalence of osteoporosis must be considered to continue to increase significantly due to the expected demographic development and environmental changes. In the diagnosis and staging of osteoporosis the three dimensional bone structure should be as important as the bone mass or the mineral content of the bone. In this study, microfragments were taken from distal radius fracture zones and investigated in Micro-CT scans. Patients in which osteodensitometry of the lumbal spine had revealed osteoporosis in were found to have significantly reduced bone mass, bone density and trabecular thickness. Trabecular fractures which were found in non-osteoporotic patients even in robust trabeculae were detected by the two-dimensional analysis in thin locations and arborizations. Despite some trabeculae turned out to be very small the differences in the histomorphometry and the quality of trabecular fractures in osteoporotic as well as non-osteoporotic patients could be visualized very good in the Micro-CT analysis. PMID- 15832578 TI - [Polymer networks as actuator and sensor systems to be used for automation of biomedical devices]. AB - Polymer networks are based on molecules which are covalently or physically connected in a three-dimensional network. In presence of an appropriate solvent these networks swell by solvent absorption to form gels. These gels, which are called hydrogels in case of water absorption, are able to change their volume by more than a hundred-fold. During the swelling or shrinking process the hydrogels perform a mechanical work. Their volume standardized working capacity can be ten times larger than that of an electromagnet. Due to their simple design, miniaturisation properties, and their ability to realize many automatic sensor and actuator functions, smart hydrogels offer new solutions in biomedical technology. PMID- 15832579 TI - Endemic alveolar echinococcosis in Southern Belgium? AB - Until now, Belgium has been considered as a low-risk country for alveolar echinococcosis. However it was recently demonstrated by necropsy series that, in some parts of southern Belgium (Wallonia), up to 51% of the red foxes (Vulpes vulpes) may be infected by E. multilocaris. The authors, working in a university hospital in southern Belgium, described in 2002 the first autochthonous Belgian case of hepatic alveolar echinococcosis. More importantly, in 2004, they diagnosed three other patients with alveolar echinococcosis. One underwent surgical resection, but two others had bilateral pulmonary involvement at time of definite diagnosis. Palliative albendazole therapy was initiated. These patients had been diagnosed with hepatic mass from unknown origin for several months. The previous experience with the first case allowed the authors to consider and to confirm alveolar echinococcosis diagnosis, made by pathology and/or serological tests and imaging. These four patients with alveolar echinococcosis were living either in the Liege or the Luxembourg province. Considering the high prevalence of E. multilocaris infection of red foxes and the recent increase of the fox population due to rabies vaccination in southern Belgium, and also the presence of E. multilocaris infection of red foxes in northern Belgium, it is likely that not only Wallonia, but also maybe the whole Belgium, may face endemic alveolar echinococcosis in the next years. PMID- 15832580 TI - Analysis of Wnt/Beta catenin signalling in desmoid tumors. AB - Desmoid tumors are fibromatous lesions occurring both sporadically and in patients with familial adenomatous polyposis (FAP). Because of the association of these tumors with the hereditary colorectal cancer syndrome FAP we set out to define the molecular events driving desmoid tumorigenesis, hypothezising these might be identical to events driving colorectal tumorigenesis. We found that whereas FAP-associated desmoid tumors are caused by germline APC mutations followed by somatic inactivation of the wild-type APC allele, sporadic desmoids are usually characterized by oncogenic mutations in the b-catenin gene, both identical molecular alterations to those found in the vast majority of colorectal cancers. Next we set out to investigate the cellular pathways activated by these mutations, and identified activation of the Wnt signaling pathway in desmoid tumors. Wnt signaling modulates expression of developmental genes and cell fate via beta-catenin, and has been implicated in many cancer types. Currently we are investigating tissue-specific downstream effectors of the Wnt pathway that might be responsible for the behaviour of these invasive fibrous tumors. Our findings also point to a role for this pathway in the regulation of normal myofibroblast proliferation and suggest novel treatments in desmoid tumors and other fibrous proliferative disorders. PMID- 15832581 TI - Capsule endoscopy examination of patients with obscure gastrointestinal bleeding: evaluation of clinical impact. AB - BACKGROUND: Wireless video capsule endoscopy (WVCE) has been reported to contribute to the diagnostic management of patients with obscure gastrointestinal bleeding (OGB). Nevertheless, clinical impact data is lacking. The aim of our study was to determine the clinical outcome of patients undergoing WVCE for OGB. PATIENTS AND METHODS: 38 patients who were referred for OGB and who underwent WVCE examination were included in this study. A questionnaire was sent to the referring doctors after the capsule investigation. The following items were investigated: the final diagnosis of OGB, treatment applied and clinical outcome. RESULTS: Data was recorded for 26 patients out of 38 (17F, 9M). The mean age was 63 years (range, 21-84). A positive finding was noted in 11 patients. Findings included small bowel lesions in 7 cases and gastroduodenal lesions in 4 cases. As a result of the capsule investigation, specific therapy was administered in 9 patients. Six of the 9 patients treated had no further anemia. Final diagnosis of OGB was in agreement with positive findings of WVCE in all these 11 patients. Among the 15 patients with a negative WVCE, 9 had a digestive lesion (3 small bowel lesions) as a final diagnosis. CONCLUSION: Patients with a positive result at WVCE had further intervention in 82% and were successfully treated in 67% of cases. We had no false positive results in this study, which suggest a high positive predictive value of WVCE. Moreover, negative WVCE had also an indirect clinical impact. PMID- 15832582 TI - Endoscopic mucosal resection: established indications, potential indications and perspectives. PMID- 15832583 TI - Longitudinal prospective study on quality of life and psychological distress before and one year after liver transplantation. AB - BACKGROUND: The impact of liver disease and medical complications on quality of life (QOL) and psychological distress before and after liver transplantation (LT) is a matter of growing interest. METHODS: In a longitudinal prospective study, perceived QOL (LEIPAD Quality of Life test) and psychological distress (Brief Symptom Inventory, BSI) were assessed in 25 cirrhotic patients when they were listed for LT and 1, 3, 6 and 12 months after LT. Patients were also evaluated for medical complications and blood levels of immunosuppressive agents. RESULTS: Overall QOL and psychological distress improved significantly and rapidly in most domains from the first month and up to a year after LT. Medical complications and immunosuppressive agents did not correlate with any changes in QOL and psychological distress after LT. When patients were divided according to liver disease etiology: 1. HCV patients listed for LT had worse QOL levels than the group of patients as a whole or the alcoholic liver disease (ALD) patients; 2. HCV patients reported a significant improvement in QOL only 6 and 12 months after LT, and still suffered more psychological distress 12 months after surgery; 3. in ALD patients, overall QOL and psychological distress improved significantly at all follow-up points after LT; 4. HCV patients reported a worse QOL and greater psychological distress 1 and 3 months after LT than the group as a whole or the ALD patients (p < 0.05). CONCLUSIONS: Liver transplantation improves QOL and psychological distress in most recipients, but not in the early stages after LT in patients transplanted for HCV cirrhosis. PMID- 15832584 TI - Genes and metals: a deadly combination. AB - Wilson's disease is an autosomal recessive disease of copper metabolism, with an estimated prevalence of 1:30000. The most common presentations of WD are liver disease and neurological disturbance. For many years the diagnosis was based on the results of several clinical and biochemical tests, for which several limitations had been reported. In recent years the developments of new techniques in genetic and molecular biology have provided useful tools in the diagnosis of Wilson's disease. However, the presence of several mutations and the fact that most patients are compound heterozygote means that the problem is not completely resolved. Chelators and zinc salts have been largely used in the treatment of WD patients with a favorable outcome, but the debate continues as to the agents of first choice. Liver transplantation is a cure for patients with decompensated liver disease but its effect on the neurological outcome is still not clear. PMID- 15832585 TI - Genetic hemochromatosis update. AB - Hereditary Hemochromatosis is an autosomal recessive disease, characterized by chronic iron overload. It is mainly due to mutations of the HFE-1 gene. In the large majority of patients, the substitution of tyrosine for cysteine at amino acid 282 (C282Y) is found at the homozygous state. Since the HFE-1 hemochromatosis identification, several other entities of iron overload have been individualized. In the present article, the frequency, penetrance and pathophysiology of HFE-1 hemochromatosis as well as various clinical presentations resulting from different mutations affecting different proteins involved in iron metabolism are described. PMID- 15832586 TI - Guidelines for the management of chronic hepatitis C in patients infected after substance use. PMID- 15832587 TI - Substance use in Belgium : prevalence and management. AB - Substance use is emerging in Belgium. The 'typical user' starts at a young age. Polydrug use has become very common. Cannabis and alcohol are the most frequently used substances among the school population. Heroin and cocaine are the most frequently injected drugs. Sharing of injecting material and paraphernalia is reported to happen in half of the subjects injecting drugs. Substance use causes a lot of adverse organic, social and psychiatric events. Management of substance use consists of information, drug-free treatment and harm reduction, including substitution and maintenance programs. The management of care in Belgium differs between the regions. PMID- 15832588 TI - The epidemiology of hepatitis C among injecting drug users in Belgium. AB - In industrialised countries, injecting drug use is currently the most important risk factor for infection with hepatitis C, resulting in high prevalence rates of hepatitis C among injecting drug users. To contain the hepatitis C epidemic major efforts should be done to prevent new infection among injecting drug users. Monitoring infection rates are crucial as it may provide feedback on the effectiveness of interventions. In this article the epidemiology of hepatitis C among injecting drug users in Belgium is briefly reviewed. More specifically the prevalence of anti-HCV antibodies, the prevalence of co-infections, the proportion of chronic HCV carriers, the distribution of genotypes and preventive measures among injecting drug users in Belgium are discussed and compared to the situation elsewhere in Western Europe. PMID- 15832589 TI - Treatment of hepatitis C viral infections in substance abusers. AB - AIMS: To examine the evidence for excluding chronic hepatitis C (CHC) patients with substance abuse from treatment with interferon (IFN) and ribavirin. METHODS: We reviewed clinical trials focussing on the treatment of chronic hepatitis C of patients with substance abuse between 2001 and 2004. Ten clinical trials concerning antiviral treatment in substance abusers were described of which six were controlled ones. There were no randomised trials. There was one controlled multi-centre trial. One trial used pegylated IFN. RESULTS: In the total group of substance abusers the sustained viral response (SVR) and the adherence was not different from control groups. In former drug users, active drug users and patients taking substitution therapy for opioid dependence the sustained viral response and adherence was not different from control populations. However, non substituted active drug users seemed more likely to be lost to follow-up. Discontinuation of treatment occurred most frequently during the first 8 weeks of therapy. Neurobehavioural changes leading to depression started in the first 8 weeks of treatment. Although follow-up periods after SVR were short, the currently described re-infection rate occurring in active intravenous drug users remains low. CONCLUSIONS: There is no evidence to withhold antiviral treatment against HCV in active substance abusers. It seems important to advise to start substitution therapy in non-substituted active drug users, increase substitution therapy dose in substituted patients and treat depression as early as possible. More prospective controlled trials on HCV treatment in active and difficult-to reach substance users are needed. PMID- 15832590 TI - Hepatitis C, interferon alpha and psychiatric co-morbidity in intravenous drug users (IVDU) : guidelines for clinical practice. AB - The evidence regarding the co-morbidity of chronic hepatitis C, psychiatric illness and intravenous drug abuse is reviewed from the literature. Also the occurrence and the treatment of psychiatric side effects during treatment with interferon in patients with a history of drug abuse are reviewed. There is insufficient evidence for a specific hepatitis C induced depression or fatigue, but a direct link between hepatitis C and cerebral dysfunction is not excluded. Immune system activation rather than drug use may explain cerebral symptoms. In HCV positive substance users anxiety and depression are more prevalent than in HCV negative substance users. During treatment with regular or pegylated (PEG) interferon depression is a frequent side effect (ca 30%) and occurs independently from pre-existing psychiatric disorders or drug abuse. A history of drug abuse per se does not increase the risk of depression as a side effect of interferon treatment. It is extremely important to monitor symptoms of depression in the early weeks of treatment and to start antidepressant treatment as early as possible. Antidepressants should be continued throughout the interferon treatment period. There are insufficient data to assess these situations in which preventive antidepressant treatment should be started before interferon treatment. Clinical judgement can, however, lead to preventive antidepressant treatment, even at subclinical levels of depression. A cut off score of > 10 on the Beck Depression Inventory before interferon treatment is associated with a higher risk of depression during treatment. Both selective serotonin reuptake inhibitors and other classes of antidepressants can be used. PMID- 15832591 TI - Methadone and buprenorphine maintenance therapies for patients with hepatitis C virus infected after intravenous drug use. AB - Heroin addiction is a chronic relapsing disease that is difficult to cure, but stabilisation and harm reduction can importantly increase the life time expectancy and the quality of life of the patient, his immediate vicinity and society in general. Currently, no proven effective pharmacological interventions are available for cocaine addiction, and treatment has to rely on existing cognitive behaviour therapies combined with contingency management strategies. Substitution therapy, however, is effective in caring for heroin addicts. Methadone is a synthetic opioid that counteracts withdrawal symptoms of heroin. Buprenorphine is a derivative of the morphine alkaloid, thebaine, and is a partial opioid agonist at the micro opioid receptor in the nervous system. A substitution treatment program effectively reduces and often eliminates heroin injection behaviour, rendering patients more socially stabilised. Reduction in the number of viral co-infections can be observed. Methadone undergoes oxidative biotransformation in the liver, but is also stored in the liver and released into the blood in unchanged form. The usual dose can be continued in patients with stable chronic liver disease, including advanced cirrhosis. In acute liver disease or acute decompensation of chronic liver disease, close clinical observation for signs of narcotic overdose or withdrawal is necessary. A modest alteration in methadone dose may be appropriate for some patients. Buprenorphine can cause liver dysfunction after sublingual and even more after intravenous administration. It is advised to follow the liver function during buprenorphine treatment and to warn the clients for intravenous use of buprenorphine. Neither methadone nor buprenorphine do influence the effect of interferon and ribavirin during the treatment of chronic hepatitis C patients. It may be necessary to increase the dosage of methadone during interferon treatment. PMID- 15832592 TI - Therapy of chronic hepatitis C in the setting of HIV co-infection. AB - Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections are major health problems world-wide. As both viruses partially share routes of transmission, co-infection is common. This is especially the case in patients infected through intravenous drug use. It has been shown that HIV accelerates HCV progression to cirrhosis. The influence of HCV infection on the natural history of HIV disease remains highly controversial. It is also known that HCV co infection increases the risk of hepatotoxicity of Highly Active Antiretroviral Therapy (HAART). These considerations as well as the improved survival of HIV patients due to HAART leads to increasing numbers of patients undergoing assessment and treatment of HCV infection. HCV treatment should be considered in stable HIV disease. Recent data indicate that HCV treatment schedules should be similar in co-infected and HCV mono-infected individuals, with pegylated interferon combined with ribavirin. For all treatment regimens published, coinfected patients had a lower sustained viral response rate compared to HCV mono-infected patients. Similar predictive factors determine the success rate. The effect of prolonging therapy to 12 months in genotype 2/3 and to 18 months in early viral responders with genotypes 1/4 needs to be assessed in further studies. PMID- 15832593 TI - Case report: acute pancreatitis induced by Clozapine. AB - Two percent of acute pancreatitis are drug induced. In the present paper, we reported the case of a 39 year-old patient with chronic-hallucinatory schizophrenia who developed symptomatic pancreatitis during the clozapine dose titration performed to reach the therapeutic range. Diagnosis of pancreatitis was suggested by clinical examination and abnormal laboratory values of pancreatic enzymes and confirmed by C-T scan and ultrasonography. The causal incrimination of clozapine in this case seems likely as all other possible causes of pancreatitis were excluded, as AP developed shortly after the introduction of the drug and as the pancreatic enzymes normalized after clozapine was stopped. No rechallenge to confirm the causal relationship was however attempted. So far, only eight cases of acute pancreatitis have been reported in association with clozapine use. Clozapine is an atypical antipsychotic drug which belongs to the chemical class of dibenzodiazepines. The mechanism by which clozapine could produce acute pancreatitis remained unclear. Nevertheless, we advocate a careful biological follow-up (measuring periodically the concentrations of amylase, lipase and triglycerides) during the treatment by clozapine. PMID- 15832594 TI - Case report: duodenal stromal tumor. AB - Tumors of the small intestine are rare lesions, but they should be kept in mind as possible causes of gastrointestinal symptoms. Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the digestive tract. A 78 year old woman complaining of abdominal pain, nausea and vomiting after meal and weight loss for three months was admitted to our clinic. On physical examination, there was only epigastric tenderness. No mass was palpated. She was anemic and total protein and albumin levels were low. Other laboratory tests were normal. A 9.0 x 7.5 cm heterogeneous mass was detected on the abdominal computerized tomography scan. Endoscopy confirmed a polypoid and vegetative mass in the second part of the duodenum. Histopathological diagnosis of endoscopic biopsy was gastrointestinal stromal tumor. Pancreaticoduodenectomy was performed. On the 11th postoperative day, relaparotomy was performed due to biliary leakage from the subhepatic drain. Biliary leakage was from the choledochojejunostomy. Choledochojejunostomy and pancreaticojejunostomy were revised. She was discharged on the postoperative 25th day. Histopathological examination of the resection specimen revealed duodenal stromal tumor. Although stromal tumors are relatively rare in the duodenum, in the case of upper gastrointestinal obstruction and anemia, this type of tumors should be considered in differential diagnosis. PMID- 15832595 TI - Development of adenocarcinoma in chronic fistula in Crohn's disease. AB - The authors report the case of a 55 yr-old woman suffering from Crohn's disease since 31 years with stricture and fistulas developed in the ileocolic junction and anorectal portion. Long-standing anorectal fistulas and stricture led to adenocarcinoma and finally fistulisation in the vagina. Diagnosis was made by perineal examination with biopsies under general anaesthesia. Treatment was first posterior pelvectomy with resection of the anterior wall of vagina. Secondarily, radiochemotherapy was administrated. The authors discuss the incidence and risk factors of carcinoma in Crohn's disease with chronic fistulas. PMID- 15832596 TI - Spontaneous regression of inflammatory pseudotumor of the liver : a case report. PMID- 15832597 TI - Therapy of acute hepatitis C with interferon-alpha2b plus ribavirin in a health care worker. AB - Hepatitis C virus can be transmitted to health care workers through needlestick accidents. In this report, the result of short-term therapy with interferon alpha2b plus ribavirin combination of acute hepatitis C in a health care worker who infected through laboratory accident was presented. The patient received combination of interferon-alpha2b (5 MU three times a week) plus ribavirin (1000 mg daily) for three months. Aminotransferase levels were normalised and clearance of HCV RNA was obtained in the first month of the therapy. After 19 months of follow-up, he had undetectable levels of HCV RNA so sustained response (clearance of HCV RNA and normalisation of aminotransferases at least six months after cessation of therapy) was achieved. According to this result, short-term therapy of acute hepatitis C with interferon-alpha2b plus ribavirin may be an alternate to others. PMID- 15832598 TI - The demographic situation in the European Union. AB - This article is an update of a similar article published in Population Trends 104 and complements the article on European-wide issues in population statistics, published in Population Trends 118. The main areas of demography are covered, namely population change, population composition, fertility and mortality. Eurostat are currently working on a new set of population projections which should be available shortly, but were not completed when this article was drafted. PMID- 15832599 TI - Replacement fertility, what has it been and what does it mean? AB - Replacement fertility is a term commonly used by demographers when referring to levels of childbearing and yet is rarely explained. It is normally presented as being around 2.1 children per woman. Continued below replacement fertility in developed countries and fertility falling in developing countries has given the concept of replacement fertility a higher profile. This article explains how replacement level is calculated and explores the concept further. Past replacement fertility levels are calculated for England and Wales. A possible alternative definition of replacement is also presented. Simple projection scenarios are used to show the effect on population of below replacement fertility, and also of postponement of fertility. The importance and implications of below replacement fertility are discussed. PMID- 15832600 TI - The creation of 'consistent areas through time' (CATTs) in Scotland, 1981-2001. AB - Small area analysis of sociodemographic change over time is often impeded by the changing configuration of census zones for each decennial UK census. Approaches for matching zone configurations exist, but these typically require population estimation techniques, which inevitably involve some error. Because of the postcode-based geographical zoning system used in Scottish censuses, it is possible to create a local-area geography which is consistent for recent censuses. Here we present a methodology used to create three sets of consistent areas through time (CATTs) that can be used for analysing comparable small area data output from the 1981, 1991 and 2001 Censuses. PMID- 15832601 TI - Cohabitation population estimates for England and Wales, 2003. PMID- 15832602 TI - Marriages in England and Wales, 2003. PMID- 15832603 TI - Marriages and divorces during 2002, and adoptions in 2003: England and Wales. PMID- 15832604 TI - Educational underachievement in children with epilepsy: a model to predict the effects of epilepsy on educational achievement. AB - In this study, we evaluated the impact on educational achievement of four characteristics of epilepsy individually and combined: epilepsy syndrome (type of epilepsy), seizure type, the frequency of epileptiform electroencephalographic (EEG) discharges, and the effect of antiepileptic treatment. Simultaneously, the effect on cognitive function and the relationship between educational underachievement and cognitive impairment were evaluated, focusing on memory, attention, speed of information processing, and intelligence level. This study was an open, controlled, parallel-group, nonrandomized clinical investigation. Eligible patients were selected when referred to our center for assessment of relationships between epilepsy and learning impairment in the years 1997 to 2001. Separately, children without neurologic deficit and without educational delay were assessed with the same tests as the children with epilepsy. This latter group is used in this study as a control group. One hundred seventy-six children with epilepsy and 113 controls were included. Gender distribution and age were comparable for the two groups. All children were in regular primary education. The children were assessed with a test battery consisting of tests for educational achievement, cognitive tests and tests for reaction time, and tests for memory and intelligence. Multivariate analysis of variance for tests of educational achievement showed a statistically significant effect for type of epilepsy (F = 4.386; P = .04), caused by the statistically lower scores for patients with localized epilepsy and symptomatic generalized epilepsy. For the reaction-time tests, a statistically significant effect for epileptiform EEG discharges (F = 3.165; P = .01) and treatment (F = 4.472; P = .001) on both vigilance tests was found, caused by patients with frequent epileptiform EEG discharges and polytherapy. Two-way interactions showed an interaction with type of epilepsy, with more patients with symptomatic generalized epilepsy having frequent epileptiform EEG discharges and polytherapy. For memory, none of the analyses showed statistically significant effects. For intelligence only for type of epilepsy, a statistically significant effect was found (F = 10.174; P = < .001). We propose a model with the type of epilepsy (epilepsy syndrome) as the dominant factor explaining educational underachievement in children with epilepsy. Such educational underachievement is most prominent for the localized and symptomatic generalized epilepsies, which suggests a dominant impact of underlying etiology (brain dysfunction or damage). These epilepsies are characterized specifically by a lower intelligence; hence, this could be the primary cognitive factor mediating between the type of epilepsy and educational underachievement. From the other factors, treatment (the use of polytherapy) and frequent epileptiform EEG discharges are associated with impaired vigilance, which could have an additional influence on educational achievement. These factors are, however, not independent of the type of epilepsy. PMID- 15832605 TI - Cerebrospinal fluid insulin-like growth factor 1 is low in acute and chronic white-matter diseases of children. AB - Insulin-like growth factor 1 increases both the number of oligodendrocytes and the amount of axonal myelin produced. The aim of this study was to see whether insulin-like growth factor 1 played a role in white-matter diseases of children. We studied insulin-like growth factor 1 in the cerebrospinal fluid of children with various white-matter diseases: (1) children with acute demyelinating events: acute disseminated encephalomyelitis (n = 5), acute transverse myelitis in multiple sclerosis (n = 1), and infarct of the medial cerebral artery causing secondary white-matter changes (n = 1), and (2) children with chronic diseases: delayed myelination (n = 3) and progressive leukodystrophies (n = 4). Insulin like growth factor 1 was determined by radioimmunoassay with commercially available kits (Mediagnost, Tubingen, Germany). We found markedly lower concentrations of cerebrospinal fluid insulin-like growth factor 1 in the patients than in the 28 age-matched control children (P < .0005). Low cerebrospinal fluid insulin-like growth factor 1 can play a role in the pathology of both acute and chronic white-matter diseases of children. PMID- 15832606 TI - Severe refractory status epilepticus owing to presumed encephalitis. AB - The severe refractory type of status epilepticus is very rare in the pediatric population. Eight children with the severe refractory type of status epilepticus owing to presumed encephalitis are described. The age at the onset of status epilepticus of the eight study children ranged between 2.5 and 15 years. Seven of the eight children presented with fever several days prior to the onset of seizures. A comprehensive clinical and laboratory investigation failed to delineate a cause for their seizures. Burst suppression coma was induced by pentothal, midazolam, propofol, or ketamine in all of the children. The mean duration of anesthesia was 28 days (range 4-62 days), but the seizures persisted in spite of repeated burst suppression cycles in all of them. Two children died. Four of the surviving children continued to suffer from seizures, and cognitive sequelae were present throughout follow-up in four children. In summary, the severe refractory type of status epilepticus of the acute symptomatic type owing to relatively mild encephalitis carries a high mortality rate and poor morbidity in terms of seizures and cognition at follow-up. PMID- 15832607 TI - Spike morphology, location, and frequency in benign epilepsy with centrotemporal spikes. AB - The literature on benign epilepsy with centrotemporal spikes reports a constellation of neurophysiologic features in selected populations with heterogeneous methodologies. The aim of this study was to determine the specific electroencephalographic (EEG) features (spike morphology, location, and frequency and associated background slowing) in a broad population-based cohort identified through EEG laboratories. The mean spike frequency in the awake state was 9.3 per minute (95% confidence interval 6.5-12.0), in drowsiness, 21.2 per minute (16.7 25.6); and in sleep, 45.6 per minute (38.3-52.8), where 60% of patients had > 40 discharges per minute. In five patients, spike train rates occupied > 80% of the sleep record, and in nine patients, they occupied 61% to 80%. An ambulatory overnight record did not add new information comparing early-onset sleep with a mean spike frequency of 37.1 per minute (27.3-46.9) with slow-wave sleep, 36.0 per minute (27.3-44.7). Patients with benign epilepsy with centrotemporal spikes have a high spike burden, which can impact on cognitive function. PMID- 15832608 TI - Childhood ischemic stroke in a nonurban population. AB - A 10-year, retrospective review of the etiology, outcome, and complications of ischemic stroke in children from a nonurban population was conducted. Twenty seven children were identified (14 boys, 13 girls), ages 1.25 to 17 years (mean 7.7 years). Etiologies included undetermined (22%), arterial dissection (19%), coagulopathy (15%), embolism (15%), moyamoya disease (11%), sickle cell disease (11%), isolated angiitis of the central nervous system or vasculitis (11%), or other known source (11%; two fibromuscular dysplasia, one L-asparaginase). More than one risk factor was present in five children. Seventeen (65%) children were anticoagulated, with no adverse events occurring. Nine children were anticoagulated initially with low-molecular-weight heparin. Other treatments included corticosteroids; physical, occupational, and speech therapy; and anticonvulsants for concomitant seizures. Follow-up ranged from 3 to 60 months (mean 17 months) and was as follows: 6 (22%) were normal, 9 (33%) had mild impairment, and 12 (44%) had moderate to severe deficits. There were no deaths. Neurologic complications included seizure (two), behavioral problems (two), and hemorrhagic conversion (one). In this population, the outcome from ischemic stroke was similar to that of other studies, with the majority of children demonstrating persistent neurologic deficits. Etiology could be determined for the majority of patients, with 19% having more than one risk factor. PMID- 15832609 TI - Screening electroencephalograms in autism spectrum disorders: evidence-based guideline. AB - The purpose of this study was to establish guidelines for the use of screening electroencephalograms (EEGs) in autism spectrum disorders to identify subclinical epileptiform activity. MEDLINE and EMBASE were searched for relevant articles. The evidence was reviewed using the methodology of the Canadian Task Force on Preventive Health Care. A priori criteria were established for article selection and for grading the quality and strength of the evidence. Seizures are common, occurring in 20 to 30% of patients based on the majority of studies. Epileptiform EEG abnormalities are present in 10.3% to 72.4% of patients and subclinical abnormalities in 6.1% to 31%. Evidence for the effectiveness of anticonvulsants and corticosteroids in reducing seizures and/or autistic symptoms is based primarily on case series and case reports, with only one published randomized trial. There is currently insufficient evidence to recommend for or against the use of screening EEGs in autistic patients. Given the frequency of seizure disorders in this patient population, a high index of clinical suspicion should be maintained for subtle symptoms of seizures. PMID- 15832610 TI - Electrophysiologic evaluation of peripheral nerve injuries in children following the Marmara earthquake. AB - The aim of this study was to investigate the clinical, demographic, and electromyographic (EMG) characteristics of 12 earthquake victims in the pediatric age group and to compare the findings with those of the adult group. Following the 1999 Marmara earthquake, 75 subjects with suspected peripheral nerve injury were referred to our EMG laboratory for evaluation. In the pediatric age group, five patients had a history of short-term temporary trauma and seven had a history of being trapped under the debris for 4 to 10 hours and sustaining long term trauma. Five patients had developed compartment syndrome and one had developed crush syndrome. The EMG examinations revealed peripheral nerve injury findings in all patients. The brachial plexus was damaged in 2 patients, and 19 peripheral nerves were damaged in 10 patients. Peroneal and posterior tibial nerves were predominantly affected. Regeneration was detected in all of the patients with brachial plexus damage at a mean follow-up of 3.5 months and in 62.5% of patients with peripheral nerve damage at a mean follow-up of 7.7 months. Being buried under the debris, compartment syndrome, peripheral nerve injuries in the lower extremities, and total axonal damage in the first EMG examinations were found to be higher in the pediatric age group. Regeneration findings were found at similar rates in both groups, with the brachial plexus being the most favorable. When it is considered that the regeneration process lasts 15 to 18 months and EMG findings mostly show pathologies in the form of neuropraxia and axonotmesis, we think that the prognosis of our patients will be good. PMID- 15832611 TI - Clinical experience with zonisamide monotherapy and adjunctive therapy in children with epilepsy at a tertiary care referral center. AB - We evaluated our clinical experience with zonisamide, a broad-spectrum antiepileptic drug, in a group of children with predominantly medically refractory epilepsy. A retrospective chart review was conducted on patients at our tertiary referral center following Institutional Review Board approval. Observers documented reports of seizure frequency, and seizure types were identified either clinically or by prior video-electroencephalography monitoring. We identified 68 patients (age range 1.9-18.1 years [median 6.9 years]; male to female ratio 1.3:1) treated with zonisamide for 0.7 to 28.9 months; at the last visit, 22% and 78% were on monotherapy and adjunctive therapy, respectively. The median duration of treatment and maintenance dose at the end of the follow-up were 11.2 months and 8.0 mg/kg/day, respectively. Seizure types included generalized (primary generalized tonic-clonic, myoclonic, tonic, atonic, absence) and partial (simple, complex, and secondarily generalized tonic-clonic seizures); 10 (15%) patients had both partial and generalized seizures. Sixteen (25.8%) patients were seizure free, although five of them were already in remission prior to starting zonisamide. Thirteen (21.0%) patients had a > 50% seizure reduction, 10 (16.1%) patients had a < 50% seizure reduction, 14 (22.6%) had no improvement in baseline seizures, and 9 (14.5%) reported having increased seizures. The latter were mostly associated with dosage alterations in concomitant antiepileptic drugs. Common side effects were central nervous system related, including behavioral or psychiatric (23.5%), cognitive dysfunction (12.0%), and sedation (10.3%). Eleven (16.2%) patients ultimately discontinued zonisamide, but only five were strictly due to side effects. Zonisamide is clinically effective against multiple seizure types in a significant proportion of children with epilepsy across a broad age range. Drug discontinuation as a result of side effects is uncommon. PMID- 15832612 TI - Epilepsy in shunted posthemorrhagic infantile hydrocephalus owing to pre- or perinatal intra- or periventricular hemorrhage. AB - Epilepsy is relatively common in infants with hydrocephalus. Its mechanism is controversial; in fact, studies on etiologically heterogeneous series are not able to clarify the mechanism generating epilepsy or to suggest effective prevention and treatment strategies. Our study is aimed at assessing the onset and evolution of epilepsy, as well as concurrent cognitive development of a homogeneous series of shunted posthemorrhagic hydrocephalus owing to pre- or perinatal intra- or periventricular hemorrhage. Forty patients were enrolled in the study. Twenty-six were patients with grade II-III intraventricular hemorrhage, 16 of whom had associated ischemic lesions. In the remaining 14 patients, a grade IV intra-ventricular hemorrhage was found. Epilepsy was observed in 27 patients. Aside from 10 cases with nonsyndromic forms of epilepsy, it was possible to define at least three different age-dependent epileptic syndromes: symptomatic neonatal location-related epilepsy with transient West's syndrome in infancy in 5 patients; West's syndrome in 8 patients; and continuous spike-waves during sleep in 4 patients. Epilepsy was significantly correlated with ischemic lesions only. Early thalamic injuries frequently evolved toward continuous spike-waves during sleep, indicating that patients with thalamic injury must be monitored to detect continuous spike-waves during sleep early. Cerebellar atrophy, in addition to epilepsy and other brain injuries, accounted for disorders of cognitive development. PMID- 15832613 TI - A pediatric neurologist's clinical experiences in Saudi Arabia. AB - From 1996 to 2002, I was head of a pediatric neurology section and helped develop a fellowship training program in Saudi Arabia. This was a most rewarding learning experience because of an abundance of neurometabolic and neurodegenerative diseases in a population with a high rate of consanguinity. In addition to inborn errors of metabolism, other prevalent disorders included late-infantile neuronal ceroid-lipofuscinosis, hyperekplexia, focal cortical dysplasias, sarcoglycanopathies, familial spastic paraplegia, demyelinating diseases, and dysmorphic syndromes. These patient experiences were a rich source of stimulation for clinical teaching and research. PMID- 15832614 TI - Megalencephalic leukoencephalopathy with subcortical cysts in two siblings owing to two novel mutations: case reports and review of the literature. AB - Megalencephalic leukoencephalopathy with subcortical cysts is a rare leukodystrophy characterized by macrocephaly and a slowly progressive clinical course marked by spasticity and cognitive decline. We report two full siblings with neuroimaging studies and clinical courses typical for megalencephalic leukoencephalopathy with subcortical cysts, in whom a pair of novel mutations in the MLC1 gene was identified. We review the current knowledge of this disorder in relation to the patients reported. PMID- 15832615 TI - Panuveitis in association with pseudotumor cerebri. AB - We report a case of an 11-year-old girl with bilateral panuveitis in association with pseudotumor cerebri. The patient underwent complete ophthalmologic, neurologic, and laboratory evaluations and was treated with therapy for pseudotumor cerebri. The patient met the diagnostic criteria for pseudotumor cerebri and also had panuveitis. Symptoms and findings of pseudotumor cerebri and panuveitis improved significantly after combination therapy of oral acetazolamide and weight reduction. The index case illustrates that pseudotumor cerebri can be associated with panuveitis. Therapy for pseudotumor cerebri might also help with the resolution of uveitis. PMID- 15832616 TI - Predominantly upper limb weakness, enlarged cisterna magna, and borderline intelligence in a child with de novo mutation of the skeletal muscle alpha-actin gene. AB - We present a 10-year-old boy from nonconsanguineous parents of Libyan (Sephardi) Jewish origin. Mild dysmorphism, hypotonia, and clubfoot deformities were noted at birth. On follow-up, he had borderline intelligence and nonprogressive muscle weakness, predominantly in the upper extremities. Physical examination revealed mild facial weakness, a bell-shaped chest cavity, kyphosis, winging of the scapula, and hypotonia of the shoulder girdle. Muscle biopsy demonstrated prominent variation in fiber size and central nuclei and numerous subsarcolemmal particles on modified Gomori trichrome stains. Electron microscopy depicted areas of disrupted sarcomeres with abnormal aggregates. Brain magnetic resonance imaging showed mild widening of the lateral ventricles and an enlarged cisterna magna. Molecular DNA analysis by polymerase chain reaction (PCR) and direct sequencing revealed a de novo heterozygous missense mutation in the skeletal muscle alpha-actin gene (ACTA1) changing codon 348 from TCG serine to TTG leucine. PMID- 15832617 TI - Pediatric refractory partial status epilepticus responsive to topiramate. AB - Topiramate was safely administered to two young children with refractory partial status epilepticus via nasogastric tube in rapid titration up to a very high total daily dose. An excellent clinical response occurred in both cases. Reaching high daily doses of topiramate within days allowed for safe discontinuation of other antiepileptic drugs in both patients. Given the high efficacy of rapidly titrated topiramate in our patients, this medication may be useful in some cases of pediatric refractory partial status epilepticus. However, more clinical studies on this therapeutic approach are needed to establish the precise role of topiramate in status epilepticus in children. PMID- 15832618 TI - Childhood case of progressive multifocal leukoencephalopathy with improved clinical outcome. AB - A 6-year-old boy who had been in remission from acute lymphoblastic leukemia for 2.5 years presented with seizures, hemiparesis, visual loss, and white- and gray matter lesions on cranial magnetic resonance imaging. The diagnosis of progressive multifocal leukoencephalopathy was established on the detection of JC virus DNA by polymerase chain reaction in brain tissue. The patient was administered several anticonvulsants, amantadine, acyclovir, and ganciclovir. He showed partial recovery. This case illustrates the possibility of long-term survival in progressive multifocal leukoencephalopathy with normal immunologic parameters. PMID- 15832619 TI - Incongruent cerebral growth in sudden infant death syndrome. AB - Sudden infant death syndrome remains a leading cause of post-neonatal mortality in developed countries. Its etiopathogenic mechanisms are unknown. In this neuropathologic study, we noticed that the weights of the brains of infants who died from sudden infant death syndrome (n = 97) were invariably heavier in comparison with those of a group of age-matched controls (n = 23) issuing from the same local population. Brain edema was not a major element, and there were no significant microscopic or macroscopic cerebral anomalies in the brains from either of the study groups. Head circumference did not show a parallel increase in infants with sudden infant death syndrome. The excessive brain weight might reflect abnormal cerebral development and could be detrimental to vital neural control. In a previous study, we disclosed cytokine overexpression in the brains of these victims. Whether increased brain weight is linked to cytokine up regulation remains, however, a moot case and merits further exploration. PMID- 15832620 TI - Depressed left ventricular contractile reserve diagnosed by dobutamine stress echocardiography in a patient with Duchenne muscular dystrophy. AB - Cardiomyopathy is a leading cause of death in patients with Duchenne muscular dystrophy. Congestive heart failure is often sub-clinical and unrecognized as a result of the severe physical limitations of this patient population. We report the case of a 16-year-old boy with Duchenne muscular dystrophy who demonstrated normal left ventricular systolic function at rest by screening transthoracic echocardiogram. This patient, however, was noted to have depressed left ventricular contractile reserve by dobutamine stress echocardiography. Dobutamine stress echocardiography can have an important role in unmasking subclinical heart failure in this patient population. PMID- 15832621 TI - A child with Friedreich's ataxia and epilepsy. AB - Epilepsy in Friedreich's ataxia is rare. We describe a 9-year-old boy with Friedreich's ataxia who had onset of symptoms in the second year of life and developed a generalized epilepsy at age 5 years. On cerebral magnetic resonance imaging, he has a subependymal gray-matter heterotopia. We suggest that his gray matter heterotopia might be related to his diagnosis of Friedreich's ataxia and that his early onset of symptoms might be related to the length of his guanine adenine-adenine (GAA) triplet repeat expansion. PMID- 15832622 TI - Unilateral or asymmetric localization of lambda waves is not a pathologic finding. AB - This article reports on three children who underwent an electroencephalographic examination. All three showed the feature of asymmetric lambda waves, which is usually interpreted as a pathologic focal phenomenon. The report documents that asymmetric lambda waves and photic driving might be normal variants, not requiring further investigation. PMID- 15832623 TI - Topiramate in the prophylactic treatment of migraine in children. AB - We present a prospective case series of 24 children diagnosed with migraine refractory to prophylactics and treated for 4 months with topiramate as the only prophylactic drug. At the final visit, the mean topiramate dose was 3.5 +/- 1.7 mg/kg/day. Nearly all patients (87.5%) reported a shorter duration of attacks, and the average pain intensity was rated as mild by 14 patients (58.3%). Eight (33.3%) patients had adverse events, none of which were serious. In our sample of pediatric patients, topiramate was effective for the prophylactic treatment of migraine in children. It was well tolerated at the doses used for titration and maintenance. Controlled trials are needed to verify the efficacy of topiramate for migraine in children. PMID- 15832624 TI - Modern imaging methods for the assessment of Langerhans' cell histiocytosis associated neurodegenerative syndrome: case report. AB - Langerhans' cell histiocytosis-associated neurodegenerative syndrome is an enigmatic manifestation, most often localized in the cerebellum and the basal ganglia. Its pathophysiologic basis is poorly understood, and effective treatment strategies are currently missing. Modern imaging modalities offer the possibility of shedding further light on this puzzling disease in a noninvasive way. We report on a 12-year-old boy with a Langerhans' cell histiocytosis-associated neurodegenerative syndrome who underwent a thorough evaluation with different modern imaging methods in addition to routine brain magnetic resonance imaging (MRI) to analyze their informative value for this condition. Additional imaging included positron emission tomography using [18F]fluorodeoxyglucose (FDG-PET), single photon emission computed tomography using [123I]2beta-carbomethoxy-3beta (4-iodophenyl)tropane and [123I]iodobenzamide, and magnetic resonance spectroscopy. The potential relevance of each method for neurodegenerative Langerhans' cell histiocytosis is discussed based on the results obtained, and a review of the literature is made. The case underlines the fact that MRI undoubtedly possesses the major role in the diagnostic evaluation and monitoring of Langerhans' cell histiocytosis-associated neurodegenerative syndrome. FDG-PET and magnetic resonance spectroscopy findings were in good correlation with the MRI results. In particular, magnetic resonance spectroscopy could provide a valuable diagnostic tool in addition to MRI in the early detection and evaluation of the neurodegenerative component of this disease. PMID- 15832626 TI - Ideal is typical. AB - A well-established finding in research on concepts and categories is that some members are rated as better or more typical examples than others. It is generally thought that typicality reflects centrality, that is, that typical examples are those that are similar to many other members of the category. This interpretation of typicality is based on studies in which participants had little knowledge about the relevant categories. In the present study, experienced fishermen were asked to give goodness-of-example ratings to familiar freshwater fish. These fishermen were of two cultural groups with somewhat different goals and ideals. Typicality was well predicted by fishes' desirability and poorly predicted by their centrality. Further, the two cultural groups differed in their typicality ratings in ways that corresponded to their different goals and ideals. For knowledgeable reasoners typicality in natural taxonomic categories appears based on ideals rather than on centrality. PMID- 15832625 TI - New treatments for denervating diseases. AB - There has been considerable recent progress in understanding mechanisms by which gene mutations cause degeneration of motoneurons and peripheral nerves. Novel therapies inspired by these insights have begun to yield promising results in mouse models of these genetic diseases. Among these have been the use of small molecules or proteins to suppress gain-of-function mutations (eg, ascorbic acid for Charcot-Marie-Tooth disease type 1A) or to restore enzyme activities that are deficient because of loss-of-function mutations (eg, treatment of Fabry's disease with recombinant alpha-galactosidase or with low-molecular-weight alpha galactosidase chaperones and treatment of spinal muscular atrophy with phenylbutyrate). Some of these therapies are already being tested in humans. Equally exciting is the prospect that small molecules and proteins will be identified that exert potent therapeutic effects in a broad spectrum of inherited and acquired motoneuron and peripheral nerve disorders. PMID- 15832627 TI - Seeing double: levels of processing can cause false memory. AB - Ordinarily, deeper levels of processing in a study session increase the accuracy of later remembering. We modified the standard levels-of-processing procedure by presenting items either once or twice in the study phase, each item being the subject of a semantic, phonemic, or graphemic question. At test, the subjects judged the frequency with which each word had occurred in the study phase. Deeper processing during encoding increased accuracy in judging twice-presented items. However, it also caused an illusion of repetition for items presented only once. The result underlines the importance of thinking of remembering as a process of evaluation and inference, rather than simple retrieval. PMID- 15832628 TI - Similarity-guided depth of retrieval: constraining at the front end. AB - Lee Brooks has done important work to show that categorization often reflects reliance on specific instances rather than on an abstract representation. His work on the advantages of using a diagnostic hypothesis to search medical stimuli has demonstrated how constraining what one looks for influences clinical reasoning. Similarly, cognitive control can be accomplished by constraining memory retrieval in ways that influence interpretation of a memory probe. Here, we report two experiments in which the similarity of study items constrained how test items were interrogated for an immediate memory test and thereby produced differences in the depth of retrieval. A novel procedure that tests foil memory was used to diagnose differences in similarity-guided retrieval depth. PMID- 15832629 TI - Judging relationships between events: how do we do it? AB - A decade ago, Allan (1993) concluded that associative models provided the best account of data generated in tasks that require human observers to judge the relationship between binary events. In the intervening years, new data have been reported that provide evidence for higher-order processes. Some have argued that these new data pose a serious threat to the viability of the associative account. The purpose of the present paper is to review this evidence and to assess the severity of this threat. PMID- 15832630 TI - Not all errors are created equal: metacognition and changing answers on multiple choice tests. AB - Two experiments investigated the role of metacognition in changing answers to multiple-choice, general-knowledge questions. Both experiments revealed qualitatively different errors produced by speeded responding versus confusability amongst the alternatives; revision completely corrected the former, but had no effect on the latter. Experiment 2 also demonstrated that a pretest, designed to make participants' actual experience with answer changing either positive or negative, affected the tendency to correct errors. However, this effect was not apparent in the proportion of correct responses; it was only discovered when the metacognitive component to answer changing was isolated with a Type 2 signal-detection measure of discrimination. Overall, the results suggest that future research on answer changing should more closely consider the metacognitive factors underlying answer changing, using Type 2 signal-detection theory to isolate these aspects of performance. PMID- 15832631 TI - From theory to application and back again: implications of research on medical expertise for psychological theory. AB - Research directed at an understanding of medical expertise is about 30 years old, and many developments in this literature parallel progress in cognitive psychology. Over the past 15 years or so, this research became much more closely identified with particular psychological theories. Initial forays into medicine were essentially direct applications of methods developed in the psychology lab to the more natural domain of medicine, with varying degrees of success. These attempts were followed by a second wave that took the psychological theories themselves more seriously in a more thoughtful application of psychological methods to the medical domain. I will argue in the present paper that the methods and theories used in the study of medical expertise have advanced to the point that there is some reverse flow and they are providing a unique and valuable perspective on the nature of thinking. PMID- 15832632 TI - Feature representations and analytic/nonanalytic processing. AB - Lee Brooks' original formulation of instance theory embedded the notion of an instance within the larger conception of a distinction between analytic and nonanalytic processing. Brooks has recently argued that features can be represented either in terms of their specific feature appearance, or in terms of the abstract information some particular instantiation embodies. This work reviews some recent studies that link reliance on different types of feature representations to different decision-making processes, and to different patterns of categorization behaviour. This in turn complicates the analytic/nonanalytic distinction, suggesting a more precise reformulation. PMID- 15832633 TI - Wanted: a new psychology of exemplars. AB - Present-day exemplar theory faces difficult challenges, and important questions have arisen about the kinds of exemplar effects and processes that are empirically supported, and about the kind of exemplar theory that could still be constructive. One question concerns whether exemplar generalization in memory and categorization is broad and collective--extending to many related exemplars stored in memory--or whether it is focused and singular--extending only to highly similar (nearly identical) exemplars. The present article considers this continuum from broad to narrow generalization. I demonstrate that in prominent memory and category tasks--tasks in which exemplar theory predicts broad generalization--generalization is in psychological reality very tightly focused. These demonstrations could ground a new, productive exemplar theory that is true to psychological process as humans conduct themselves in memory and category tasks. This new psychology may actually reprise the traditional exemplar theory that predated our sophisticated, mathematical exemplar models. PMID- 15832634 TI - Abstract analogies and positive transfer in artificial grammar learning. AB - Following Brooks and Vokey (1991), we show that positive transfer to new items generated from an artificial grammar in which the vocabulary has been changed from training to test can be based on "abstract analogy" to specific training items (specific similarity) rather than abstraction of a grammar and symbol remapping rules, even with remapping unique to each test item. The results confirm that transcendence over symbols provides no support for the implicit learning of abstract structure. Ironically, they also show that the effect of specific similarity does not depend on surface characteristics of the items, but the residual effect of grammaticality does. PMID- 15832635 TI - The blossoms and the weeds. AB - As attested by the articles in this issue and the papers given at the festschrift, I have had the privilege of working with a talented group of people. In this article, a critique is given on the bouquet of themes that have bound many of us together, sorting the blossoms from some of the weeds. A brief description is then given of how these themes are working out in our current research on categorization. PMID- 15832636 TI - Burns and scalds--epidemiology and prevention in a developing country. AB - BACKGROUND: Burns and scalds are common presentation to many of our health institutions. Most of these injuries are preventable. Many of the patients however end with severe morbidity or even death. People affected are mostly of poor socioeconomic status. The cost of managing these injuries is high. Coupled with this are poor facilities in most parts of the developing countries like Nigeria and insufficient personnel to take care of this group of people. Study of the epidemiology as well as suggestions on preventive measures are therefore overdue and hence this publication. METHODS: A review of the literature on this subject with emphasis on Nigeria was done bearing in mind the epidemiology and prevention of burn. Literature search was done using the medline as well as local journals. RESULTS: Causes of burn injuries are many in the developing countries. Most of these are however preventable. Some preventive measures have been suggested by workers in this part of the world. Some measures that have worked in reducing burn injuries in some advanced countries and that may be appropriate to our situation are noted and are also suggested in this paper. CONCLUSION: Prevention of burn injuries, based on the epidemiology of burn in developing countries, remains a major way of reducing the current spate of morbidity and mortality in our patients. PMID- 15832637 TI - Management of blunt abdominal trauma in Maiduguri: a retrospective study. AB - BACKGROUND: Blunt abdominal trauma (BAT) presents special surgical problems and is a continuous challenge to the surgeon's ability to make an early diagnosis and provide adequate treatment. The aim of this paper is to study retrospectively the patterns, surgical management and outcome of treatment among patients with BAT. METHODS: The records of BAT patients managed at the University of Maiduguri Teaching Hospital (UMTH) between January 1998 and December 2000 were studied. Information about the age, sex, cause of injury, main investigations, diagnostic procedures and treatment of these patients were extracted from the files and analysed. RESULTS: There were 48 males and 10 females. The mean age was 25.17 years, and the patients ranged between 8 and 50 years in age. Incidence of BAT was most prevalent in the 20-40 year age group (68.96%). Road traffic injuries (RTI) accounted for majority of injuries. The main findings at presentation were abdominal pain, abdominal tenderness and hypovolaemic shock. The spleen and the liver were the most frequently injured solid organs while the small intestine was the most frequently injured hollow viscus. 28 of the patients had associated injuries involving the extremities, chest and head. Fifty patients underwent exploratory laparotomy where definitive management depended on findings. Complications included wound infection, pneumonia, and intra-abdominal abscesses. The mortality rate was 17.24%. Non-operative management was possible in 8 patients. CONCLUSION: BAT affects mainly the young people below 40 years of age. Definite preoperative diagnosis and the decision to operate was based on clinical assessment and some diagnostic procedures. 20.68% of patients were subjected to non-therapeutic laparotomies. This could be reduced by routine use of modern imaging techniques and thus avoiding unnecessary laparotomies and promoting non operative management of BAT. PMID- 15832638 TI - Intestinal malrotation: presentation in the older child. AB - BACKGROUND: The clinical diagnosis of intestinal malrotation in the older child is not always easy because of its non-specific presentations. The aim of this study was to determine the pattern of presentation of malrotation in older Nigerian children. METHODS: The clinical, radiological and operative records of all the children aged 2 years or above, managed for malrotation at the Jos University Teaching Hospital between March 1992 and December 2002 were retrospectively reviewed. RESULTS: There were 9 patients, with a median age of 5 years (range: 3-14 years). The commonest complaint was intermittent colicky abdominal pain in 9 (100%), followed by recurrent vomiting in 8 (88.9%), haematemesis and constipation each in 5 (55.6%) and repeated episodes of bloody stools and diarrhoea. Other features included abdominal distension in 5 (55.6%) and failure to thrive in 4 (44.4%). Preoperative diagnosis was possible only in 3 patients, through the use of barium meal. Operative findings included obstructing bands of Ladd, partial volvulus and mesocolic hernias. Surgery promptly and satisfactorily relieved the symptoms. CONCLUSION: The diagnosis of intestinal malrotation should be considered in any child with prolonged history of recurrent colicky abdominal pain, vomiting or diarrhoea, especially if there is associated history of failure to thrive. Surgical intervention provides satisfactory relief of symptoms and should be implemented as soon as the diagnosis is made. PMID- 15832639 TI - Awareness and attitude of antenatal clients towards HIV voluntary counselling and testing in Aminu Kano Teaching Hospital, Kano, Nigeria. AB - BACKGROUND: Mother to child transmission accounts for the majority of HIV infections in children in the developing countries. This study assessed pregnant women's knowledge of HIV/AIDS, awareness and attitudes towards Voluntary Counselling and Testing (VCT) in a teaching hospital in northern Nigeria. METHODS: A pre-tested structured interview questionnaire was administered on a cross-section of 210 antenatal clients in Aminu Kano Teaching Hospital, Nigeria. RESULTS: All respondents were aware of HIV/AIDS. Fifty seven percent had good knowledge, 32% had fair knowledge and the remaining 11% had poor knowledge of the infection. Most respondents were aware of VCT through health workers, mass media and friends. Similarly, most respondents (81.0%) approved of VCT, 13.0% disapproved of it and the remaining (6%) was undecided. The main reasons for disapproval were; fear of stigmatisation, isolation and effect on marriage security. Those that had tertiary level of education were three times more likely to accept VCT compared to those with lower levels of education (O.R=3.2, 95% confidence interval =1.3-8.0). CONCLUSIONS: Although the awareness of VCT for HIV was quite high with most antenatal clients harbouring positive attitudes towards it, there is a need to intensify health education to convince the remaining minority who are still sceptical or ignorant of the benefits of VCT. PMID- 15832640 TI - Some haematological parameters in human immunodeficiency virus (HIV) infected Africans: the Nigerian perspective. AB - BACKGROUND: Haematologic abnormalities are among the most common manifestations of advanced human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS). A specific diagnosis of cause, severity and mechanism of cytopenia should be sought because of specific treatments or intervention may be indicated for its correction. This study was to determine some haematological parameters in HIV/AIDS infected Nigerians. METHOD: One hundred HIV/AIDS infected previously antiretroviral naive adult Nigerians, aged 18-58 year (males 47 and females 53) consisting of 88 symptomatic and 12 asymptomatic patients recruited into the antiretroviral pilot project in the Haematology department of the University of Port Harcourt Teaching Hospital between June 2002 to July 2003 were studied. Haematological parameters of hemoglobin, white cell count, platelet count, erythrocyte sedimentation rate and differential leucocyte count were determined. Data was analyzed using a multipurpose statistical package version 9 SPSS. RESULT: The mean haemoglobin was 10.25 +/- 1.97 g/dl (range 6.31-14.2 g/dl), severe anaemia occurred in 80% of subjects while 20% were non-anaemic. Haemoglobin values was found positively correlated to lymphocyte count (r = .319, P = 0.01) and inversely correlated to ESR (r = -.343, P = 0.01) and neutrophil count (r = -.343, P = 0.01). Red cell morphology was variable with majority normochromic and normocytic (64%) and 36% showing hypochromia and anisopoikilocytosis. The mean total WBC count was 4.51 +/ 1.82 x 10(9)/l (range 0.9-8.2 x 10(9)/l). Leucopaenia occurred in 10/100 (10%) of study population. Total white cell count showed a significant inverse correlation to lymphocyte count (r = -.326, P = 0.01). The mean neutrophil count was 2.32 +/- 1.58 x 10(9)/l (range 0.00-5.48). Neutropaenia occurred in 24% of subjects. Neutrophil count showed a significant positive correlation with total white cell count (r = .314, P = 0.01) and a negative correlation with lymphocyte count (r = -.982, P = 0.01). Striking eosinophilia occurred in 3% of subjects. The mean platelet count was 170.07 +/- 49.03 x 10(9)/l (range 72-158 x 268 x 10(9)/l). Thrombocytopaenia occurred in 10/100 (10%) of subjects. The mean erythrocyte sedimentation rate was higher than that in healthy Africans (mean 78.87 +/- 39.33mm fall/hour (range 0.2-158mm fall/hour). CONCLUSION: Observation from this study will serve as a guide to clinicians caring for HIV patients in taking rational decision on haematological complications of HIV infection. This constitutes further evidence of the need for routine monitoring of some haematological parameters of HIV/AIDS infected Africans and before commencement of highly active antiretroviral therapy to ensure that mortality and morbidity are minimized and quality of life optimized. PMID- 15832641 TI - Pre-marital HIV testing in couples from faith-based organisations: experience in Port Harcourt, Nigeria. AB - BACKGROUND: This descriptive cross-sectional study was conducted among prospective couples referred from Faith-Based Organisations in Port Harcourt, Nigeria for pre-marital HIV screening. The study sought to establish the sero prevalence of human immunodeficiency virus (HIV) in this peculiar study group. METHODS: A total of 84 healthy heterosexual couples who required pre-marital HIV screening were tested between January 2000 and December 2003 using a Double ELISA confirmatory test of Immunocomb and Genscreen HIV I&II Kits. RESULTS: Amongst the 168 individuals tested, 35 (20.8%) were found positive. Seroprevalence was significantly higher among females 23 (27.4%) compared to males 12 (14.3%). Infection rate was highest in the 25-29 years group (29.7%, n=22) and lowest in those of 35-39 years (6.1 %, n=2), though this difference was not statistically significant (p-value=0.058). Infection rate was significantly higher among females (p-value=0.036); among prospective couples from Orthodox churches (p value=0.021); couples with prolonged courtship (>6 months) (p-value=0.0001); couples with history of premarital sex (p-value=0.0001); and couples with history of cohabitation (p-value=0.0001). CONCLUSION: Our findings prompt a wake-up call for faith-based organizations (FBOs) to urgently initiate or be more receptive of measures that emphasize behavioural and social changes amongst members. Government and non-governmental organizations should organise capacity building training for religious based organizations to enable them cope with the challenges of HIV/AIDS. The outcomes of this study further underscores the value of voluntary counselling and confidential HIV testing and especially pre- and post-test counselling as the basis of pre-marital HIV testing. PMID- 15832642 TI - Haematological parameters in prospective nigerian blood donors rejected on account of anaemia and/or microfilaria infestation. AB - BACKGROUND: Worldwide, prospective blood donors are screened for blood transfusion-transmissible diseases. In addition, predonation fitness requires adequate haematocrit and, in the tropics, negative screening for microfilaria that may precipitate allergy. The high prevalence of anaemia and microfilaria, though treatable, has contributed to the dearth of eligible blood donors. This study aims to characterize anaemia in prospective blood donors rejected for anaemia and find haematological effects of microfilarial infestation in prospective blood donors. METHODS: This prospective study was carried out from 1st of August to November 30th, 2002 at the blood transfusion unit of the Haematology Department of Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC), Ile-Ife. All consenting prospective blood donors that were rejected for anaemia and or microfilaria infestation during the study period were studied for their age, sex and haematological profile. A randomly selected population of successful donors was similarly studied. RESULTS: Sixty rejected prospective blood donors (5.2% of the total screened)) were studied. Forty-five (75%) of them were rejected for anaemia alone, 10 (16.7%) for microfilaria alone and 5 (8.3%) for both anaemia and microfilaria. The mean ages of those rejected were 33.3(+/- 9.9) years for anaemia alone, 29.9(+/- 8.5) years for microfilaria alone and 35.4(+/- 8.3) years for those with anaemia and microfilaria combined. The mean age of the successful group was 28.9(+/- 8.5) years. Of the 60 rejected subjects, 53 were males while 7 were females. Blood film of the anaemia group revealed features suggestive of iron deficiency anaemia (hypochromic microcytic cells) in 60% of them. The white cell count (WCC) was significantly increased in the microfilarial group compared to others and it revealed lymphocytosis and eosinophilia. CONCLUSION: The importance of these findings have been discussed in line with the existing literature. The need for intensive health education to encourage voluntary donation and promote the interest of females in blood donation is emphasized. PMID- 15832643 TI - Impact of scarcity of automobile fuel on attendance at a referral centre for tuberculosis in Nigeria. AB - BACKGROUND: Tuberculosis in its pulmonary and extra-pulmonary form is still a common finding in the developing world. This study was to investigate the impact of a period of scarcity of automobile fuel in the Niger-Delta area of Nigeria on attendance at a Tuberculosis Referral Centre in Igbogene, Yenagoa, Bayelsa State. METHODS: Data was collected on attendance before, during and after the scarcity. This Tuberculosis and Leprosy Control Centre is run by a German non-governmental organization. (NGO), and the State's Ministry of Health. No user fee is charged. RESULTS: During the period of scarcity of automobile fuel, the total 12-months attendance of males decreased to 25.1% of that before the advent of scarcity. Attendance rose in the post-scarcity period, but reached only 78.5% of the pre scarcity level. For females, the 12 months attendance during the scarcity period reduced to 75.3% of the pre-scarcity level. The 12-months attendance by females rose in the post-scarcity period to 109.2% above the pre-scarcity level. There was no evidence of an increase in attendance over the course of the period of automobile fuel scarcity among either males or females. CONCLUSION: The scarcity of automobile fuel which usually translated to an increase in cost of transportation probably increased the number of untreated tuberculosis patients in the population with potential serious long term health implications. The provision of vehicles as well as facility for storage of automobile fuel to this referral Centre by either the NGO or the Ministry of Health is suggested. This would allow the Centre carry out domiciliary service to these patients. PMID- 15832644 TI - Drug compliance among hypertensive patients in Port Harcourt, Nigeria. AB - BACKGROUND: Hypertension contributes significantly to cardiovascular morbidity and mortality. Adequate blood pressure control would therefore reduce cardiovascular morbidity and mortality, however adequate blood pressure control requires good treatment compliance. METHODS: One hundred consecutive patients aged 30-79 years attending the cardiac clinic of the medical out-patients clinic of the University of Port Harcourt Teaching Hospital were directly questioned about compliance with their antihypertensive drugs and results entered into the questionnaire designed for the study. RESULTS: Compliance was good in sixty percent (60%) of respondents, fair in twenty nine percent (29%) and poor in eleven percent (11%). Compliance was also found to be good in sixty-seven percent (67%) of patients with tertiary education, good in forty one percent (41%) of those with primary education. Compliance was good in seventy four percent (74%) of those taking one drug, good in only thirty three percent (33%) of those taking four drugs. Patients taking single daily dose drugs had good compliance in seventy percent (70%), twice daily dosing had good compliance in fifty five percent (55%) and among those taking thrice daily dosage, compliance was good in only seventeen percent (17%). CONCLUSION: The study shows that good compliance with anti-hypertensive therapy is best achieved with monotherapy given as single dosage. It also shows the role of education in the level of compliance. PMID- 15832645 TI - Assessment of dietary knowledge, practices and control in type 2 diabetes in a Nigerian teaching hospital. AB - BACKGROUND: In spite of the importance of diet in the management of diabetes mellitus (DM), diabetics are often unaware of its place in ensuring good glycaemic control. Consequently, compliance and adherence with dietary advice remain poor among diabetics. The standard of practice of dietary therapy for DM among physicians is also low. The aim of this study therefore was to assess the dietary knowledge, practices and control of type 2 DM in Obafemi Awolowo University Teaching Hospitals' Complex, Ile-Ife, Nigeria. METHOD: All 33 type 2 diabetes patients that attended the hospital over a three month period were studied to assess knowledge of DM, dietary practice and control. Percentage perceived, correct and accurate knowledge as well as practice were used for comparative analysis. RESULTS: All 33 subjects had truncal obesity and needed to lose weight. This was moderately severe in 60% of subjects. About 52% received dietary advice. The latter had a significantly higher mean dietary knowledge score than those without dietary advice. Significantly higher mean knowledge scores seemed to be associated with better dietary practices and better glycaemic control. Overall, dietary practices improved significantly following diagnosis and counselling. A significant proportion of subjects increased their use of food with low glycaemic index (legumes 48.5%, cereals 90.9%) following diagnosis. CONCLUSION: The findings further emphasize the importance of structured dietary advice and dietary control in Type 2 diabetes. In settings where dieticians are scarce, physicians managing diabetic patients must be skilled in the dietary management of the condition and show commitment to it. PMID- 15832646 TI - Care-seeking behaviour of caregivers for common childhood illnesses in Lagos Island Local Government Area, Nigeria. AB - BACKGROUND: The objectives were to estimate a 2-week prevalence for common childhood illnesses targeted by the National Integrated Management of Childhood Illness (IMCI) Initiative and to determine care-seeking behaviour of mothers/caregivers for these illnesses. METHODS: A cross-sectional community survey was conducted in the Lagos Island Local Government Area of Lagos State. Eligible children aged 0-5 years and their mothers/caregivers were recruited into the study by cluster-sampling technique. RESULTS: Four hundred and ninety-five children of 450 mothers were identified. Of these, 426 (86.1%) children belonging to 390 mothers/caregivers had symptoms suggestive of malaria, acute respiratory infections, diarrhoea and measles. Care was sought Outside the home at the onset of symptoms for 280 (65.7%) while 146 (34.3%) were treated at home. Of the 280 who were taken for care outside, 23 (8.2%) were taken for care at the onset of illness while the others were taken for care after an attempt at self-treatment (68.6%), use of traditional medicines (12.5%) and provision of traditional home care (10.7%). Only 65 (23.2%) of the children were taken for care within 24 hours of perceived onset of the illness. CONCLUSION: IMCI targeted childhood illnesses were prevalent and poor maternal response that could contribute to complicated outcome was identified. PMID- 15832647 TI - Histopathological characteristics of female breast carcinomas seen at the University of Port Harcourt Teaching Hospital, Port Harcourt Nigeria. AB - BACKGROUND: Breast carcinoma is an unpredictable disease in the sense that some patients may present with relatively early disease and die of widespread metastases within six months to one year, while others present with fairy advanced disease and yet survive longer. The aim of this study was to characterize breast carcinomas into prognostic groups using histological features. METHODOLOGY: A total of 269 breast carcinomas in females received during a five-year period (January 1991 to December 1995 inclusive) were analyzed. Archive records (request forms and microscopic slides) of all breast malignancies diagnosed in the department of Anatomical Pathology of University of Port Harcourt Teaching Hospital were retrieved and evaluated with respect to histological type, histological grades, and mononuclear cell infiltration within the primary tumour. The modified Bloom-Richardson system of microscopic grading of breast carcinoma was used. RESULTS: The majority of carcinomas showed scanty or absent tubule formation (43.1%) and 59% of cases showed numerous mitotic figures. Forty-nine percent of carcinomas show poorly differentiated or anaplastic nuclei. The final tumour grade (FG) which is the summation of tubular pattern, nuclear pleomorphism and mitotic grades of each of the tumours reveals that 53% fall in FG3, 26% in FG2while only 21% fall in FG1. The majority of the breast carcinomas in this study, 58%, show scanty or absent mononuclear infiltration. CONCLUSION: The majority of breast carcinomas in this study are poorly differentiated invasive ductal carcinoma, "not otherwise specified" which exhibit high proliferation ratio and are associated with poor host cellular immune reaction. These attributes translate to poor prognosis. PMID- 15832648 TI - Pancreatic pseudocyst in Ile-Ife revisited: 5 more cases. AB - BACKGROUND: Pancreatic pseudocyst is a major health problem in the developed countries and its incidence is noted to be getting higher among the Caucasians as a result of better diagnostic techniques. This retrospective study was done to review the cases of pancreatic pseudocyst seen between 1991 and 1999 at the Obafemi Awolowo University Teaching Hospitals, Ile-Ife, Nigeria. The 5 cases seen over this period of study were used to illustrate its rarity. METHOD: The case notes of the patients managed for pancreatic pseudocyst during the period under review were studied and analysed to evaluate the different modes of presentation, investigations done and the modes of treatment. RESULTS: The age range of the patients varied between 23 and 70 years with three of them being male while two were female. The most common presenting symptom and sign were abdominal pain and abdominal mass. Abdominal ultrasonography, chest X-ray, upper gastrointestinal endoscopy and laboratory blood tests were the investigative procedures carried out. The treatment modalities were either conservative or surgical (operative). One patient had spontaneous resolution of the pseudocyst, 2 had external drainage while the remaining 2 had cystogastrostomy. Two of the patients responded well to treatment and were being followed up in the Surgical Out Patient Clinic, one died within 24 hours of surgery while the other developed diabetes mellitus 3 months post surgery and was referred to the physicians for follow-up. CONCLUSION: Though pancreatic pseudocyst remains uncommon in Nigeria, ultrasonography can play an essential role in assisting diagnosis while successful management of this condition is still possible in the absence of current endoscopic techniques of drainage. PMID- 15832649 TI - Pattern of childhood psychiatric disorders in Port Harcourt, Niger-Delta region of Nigeria (January 1999-December 2002). AB - BACKGROUND: Psychiatric disorders in children are uncommon but remain a great source of anxiety to parents. In view of the paucity of such studies in Eastern Nigeria, this study has become imperative. METHOD: All children seen in the psychiatric out patient clinics, psychiatric wards and other departments were studied. Diagnosis was made with criteria of the Diagnostic and Statistical Manual (DSM) IV of 1994. RESULTS: Out of a total number of 1,645 psychiatric cases seen, 411 were children representing 25%; of these 411 cases, 235 were females (57.2%) and 176 were males (42.8%). Depressive illness constituted 35%, anxiety disorders 30%, drug related disorders 17%, conduct disorders 9%, and others 9%. Gender differences were noted in all the conditions listed above except for enuresis, mental retardation, epileptiform psychosis and schizophrenia. CONCLUSION: Government Health Policy to protect the mental health of our children so as to guarantee their full mental development should be developed and implemented. PMID- 15832650 TI - Marjolin's ulcer: report of 4 cases. AB - BACKGROUND: Marjolin's ulcer is a rare and often aggressive cutaneous malignancy that arises in a previously traumatized or chronically inflamed skin particularly after burns. The aim of the study is to highlight the surgical importance of this ulcer and to emphasize the necessity of closely monitoring unstable scars of chronic wounds especially post burns wounds. METHOD: A report of four cases of Marjolin's ulcer seen over a fifteen-year period at the University of Port Harcourt Teaching Hospital is presented with a review of the literature. RESULTS: Four cases were seen over a period of fifteen years. There were two females and two males aged 23, 65, 46, 19 years respectively. They all sustained injuries over 20 years prior to being seen at the surgical clinic and presented with ulcers that had features of malignancy. The two females had their ulcers excised and grafted, and were discharged from the hospital tumour free. The two males presented to the hospital very late with malignant ulcers, which could not be resected and they took their own discharge from the hospital. CONCLUSION: Marjolin's ulcer is uncommon in this environment but with the increasing incidence of petrochemical burns due to the oil industry in the environment, incidence of Marjolin's ulcer might increase. PMID- 15832651 TI - Post-thyroidectomy metastatic thyroid cancer. AB - BACKGROUND: The objective of this paper is to highlight the importance of tissue biopsy for histopathological diagnosis. METHOD: A report of a 46-year-old farmer with post-thyroidectomy metastatic thyroid cancer. RESULT: A post-thyroidectomy patient presenting with extensive skull and lumbar spine metastasis, paraparesis, huge occipital swelling and good healed scar without tumour residue. The thyroidectomy specimen was not subjected to histopathological diagnosis. Fine needle aspiration cytology of the occipital swelling revealed metastatic follicular carcinoma. Patient was placed on thyroxine 0.5mg daily with great improvement of lower limb muscle power. CONCLUSION: These bony metastases could have been reduced if proper investigation was done. There is need for continuous medical education for all clinical medical personnel. PMID- 15832652 TI - Dog bite of the face in an adult Nigerian--a case report. AB - BACKGROUND: Dog bite of the face is uncommon in adults. This is, in fact very rare in our environment and presentation of this single case to our centre led to our desire to discuss both the presentation and management of the patient in this paper. METHODS: An adult female Nigerian who had dog bite to her face involving both lips and the commisure is presented. The folder of the patient and the operation note were the sources of the information. RESULTS: She had minimal debridement of the wound with Abbe-Estlander flap cover of the wound. The dog was also placed under surveillance for a period of 2 weeks. The wound healed perfectly with no complication. The involved dog remained healthy 2 weeks after the injury. CONCLUSION: Dog bite of the face, though uncommon, can present in an adult. Sometimes the challenges of reconstruction of such defect could be great. Wound management with minimal debridement and closure gives good results on the face. PMID- 15832653 TI - Glomus tumour--the report of a case in an adult Nigerian. AB - BACKGROUND: Glomus tumour, a relatively common lesion in Caucasians is reported to be rare in blacks. METHOD: A case of glomus tumour is reported in an adult male Nigerian to highlight its distinctive clinical presentation and the literature reviewed. RESULT: A 60-year old man presented with a painful skin nodule on the ventral surface of his right forearm. The pain was far in excess of the size of the lesion. There was however no relief from previous medications. Following complete excision of the lesion, pain relief was dramatic. Histopathological examination of the lesion showed sheets of round to polyhedral glomus cells and elongated endothelium lined vascular channels diagnostic of glomus tumour. CONCLUSION: Severe pain is a very important feature of glomus tumour of the skin. This is relieved by complete excision of the lesion. PMID- 15832654 TI - Proposal for lateral malleolar reconstruction in fibula loss proximal to the syndesmosis. AB - BACKGROUND: High energy ankle injury can be devastating. Fibula loss proximal to the syndesmosis is a management challenge. The aim of this paper is to show that simple fusion of the floating lateral malleolus to the tibial plafond can restore the ankle mortice integrity. METHOD: A case report of a patient that had spontaneous fusion of the lateral malleolus to the lateral tibial plafond after a high energy injury and subsequent sepsis. RESULT: Uneventful union of the lateral malleolus to the lateral tibial plafond. CONCLUSION: Ankle fusion, bone lengthening procedures are time consuming and costly. A simple fusion of the lateral malleolus to the tibial plafond will restore stability to the ankle joint. PMID- 15832655 TI - [Cardiovascular monitoring and its consequences in oral surgery]. AB - The increasing number of high-risk patients in oral surgery makes it necessary to monitor the patient's general condition while operating. The surgeon needs a simple technical procedure to detect an impending clinical incident in time. From June 2002 to June 2004, 3012 patients underwent surgery in local anesthesia and were monitored with Colin BP 306 compact monitor. Heart rate, blood pressure, and oxygen saturation were checked non invasively during surgery. 17 times the operation was cancelled due to blood pressure problems. The treatment was cancelled twice due to cardiac arrhythmia. The average of the 19 patients' age was 63.5. Monitoring patients during surgery enables the surgeon to recognize high-risk situations immediately before the operation, to detect problem situations early and to avoid them, to work more calmly because of the enhanced security factor. PMID- 15832656 TI - [Clonidine in oral medicine. Literature review and our experience]. AB - There is a large experience in premedication with clonidine (Catapresan) for general anaesthesia. Clonidine is an alpha2-adrenoceptor agonist exerting central sympatholytic effects. Premedication with clonidine blunts the stress response to surgical stimuli and the narcotic and anaesthetic dose can be reduced. Furthermore, perioperative myocardial ischemic events can be prevented by preoperative application of clonidine. Oral clonidine at a dose of 1.5-2 microg/kg BW combines the advantages of benzodiazepines and morphine: anxiolysis, sedation and analgesia with stable hemodynamics and respiration. Clonidine does not have morphine related side effects such as nausea and vomiting. Doses of up to 5 microg/kg BW have been administered to young and healthy patients preoperatively in dental and maxillofacial surgery without significant side effects. However, Clonidine 2 microg/kg BW should be an adequate oral premedication dose for young and healthy patients scheduled for dental and facial surgery procedures performed under local anaesthesia in the ambulatory setting. In elderly patients clonidine 2 microg/kg BW administered orally should not be exceeded to avoid excessive hypotension and sedation. Bradycardia is a contraindication for the use of clonidine. PMID- 15832657 TI - [Controlled orthodontic extrusion with subsequent implantation]. AB - Controlled orthodontic extrusion constitutes a non-surgical technique to increase alveolar bone material. Prosthetic reconstruction alone frequently does not achieve sufficient long-term aesthetics. An adequate implantation bed is indispensable to meet aesthetic demands. Controlled orthodontic extrusion represents an alternative to pre-implantological augmentation of the vertically reduced alveolar process and its soft tissue. PMID- 15832658 TI - Transformation trauma in Buffalo City. PMID- 15832659 TI - Workplace-acquired HIV--ignorance pervasive. PMID- 15832660 TI - Asian tsunami--a lesson in misplaced ubuntu. PMID- 15832661 TI - Traditional medicines--no clear or present danger. PMID- 15832663 TI - High five. PMID- 15832662 TI - A case for further strengthening of the national health system in South Africa. PMID- 15832664 TI - Does the 'power of the purse' influence the fair distribution of limited resources? PMID- 15832665 TI - HIV/AIDS in Africa--a role for the mental health practitioner? PMID- 15832666 TI - Inhaled steroid use in adult asthmatics--experience at a primary health care centre. PMID- 15832667 TI - Pentoxifylline for heart failure: a systematic review. AB - BACKGROUND: Recent trials have indicated a beneficial effect of pentoxifylline on measures of inflammation and markers of cardiac dysfunction in people with heart failure. However, it is uncertain whether pentoxifylline should be used routinely in the management of heart failure. OBJECTIVE: To determine the effectiveness of pentoxifylline in heart failure. DESIGN: Systematic review of randomised controlled trials. METHODS: We searched MEDLINE (1 January 1966 - 20 November 2004), the Cochrane Controlled Trials Register (issue 4, 2004), and reference lists of related papers, for randomised controlled trials of pentoxifylline in the treatment of heart failure. Prospective, randomised, double-blind controlled trials were sought for inclusion in the study. The two reviewers independently assessed trial quality and extracted data, which were analysed using RevMan statistical software. The following outcome measures were evaluated: (i) New York Heart Association (NYHA) functional class; (ii) left ventricular ejection fraction (LVEF); (iii) frequency of hospitalisation; and (iv) death from all causes. RESULTS: Four studies with a total of 144 participants met the inclusion criteria. Statistical pooling (or meta-analysis) was not performed owing to the significant clinical heterogeneity and differences in reporting of the outcomes in the included studies; instead, the trials were analysed separately for the outcomes of interest. The four studies tested the use of pentoxifylline versus placebo in patients with heart failure of varying aetiology (idiopathic dilated cardiomyopathy, 3 studies; ischaemic cardiomyopathy, 1 study). In 2 of the idiopathic dilated cardiomyopathy studies, patients were classified as NYHA class II or III, while the study population in another idiopathic cardiomyopathy study was in NYHA class IV. The trial of patients with ischaemic cardiomyopathy included patients in NYHA functional classes I - IV. The use of pentoxifylline was associated with significant improvement in symptoms (i.e. NYHA functional class) and cardiac function (i.e. LVEF) in 3 out of 4 studies. The beneficial effect on symptoms of heart failure and cardiac function was seen in all grades of severity of heart failure and in patients with ischaemic and idiopathic dilated cardiomyopathy. All 4 studies showed a trend towards reduction of mortality, but this effect was not statistically significant. The effect of pentoxifylline on the frequency of hospitalisation has not been tested in randomised controlled trials. INTERPRETATION: Pentoxifylline may have a beneficial effect on NYHA functional class, ejection fraction and mortality in heart failure, but published trials are too small to provide conclusive evidence. There is a need for large, placebo-controlled trials of pentoxifylline in heart failure, involving a diverse group of patients with regard to cause and severity of heart failure. PMID- 15832668 TI - Can medical scheme reform lead to fairer distribution of limited resources? A funding perspective. PMID- 15832669 TI - Amiodarone-induced thyroid dysfunction. AB - BACKGROUND: Little is known about the frequency of thyroid dysfunction (TD) associated with amiodarone therapy in southern Africa. OBJECTIVES: To determine the incidence of TD in a cohort of patients initiated on amiodarone therapy at a cardiac clinic in Cape Town, South Africa, believed to be an iodine-replete area. PATIENTS: Pharmacy records were used to obtain the names of patients who received amiodarone between November 1999 and December 2002. RESULTS: The sample size was 194, but data analysis was limited to the 163 patients for whom there were complete data. The mean age +/- standard deviation (SD) was 59.0 +/- 15.0 years (range 22 - 89 years). There were 67 female and 96 male patients. The indications for amiodarone therapy were supraventricular tachycardias (N = 102, 62.6%), ventricular tachycardia (N = 55, 33.7%), and prophylaxis against tachycardias (N = 3, 1.8%). The indication was uncertain in 3 patients (1.8%). The median duration of amiodarone treatment was 679.0 days (quartile deviation (QD) 1172 days, range 3 - 6425 days) in the whole cohort. The median duration of amiodarone therapy until new TD was 943 days (QD 1185 days), significantly longer than in patients who remained euthyroid (547 days, QD 1135 days) (P = 0.05). There were 45 new TD cases (27.6%): 11 patients (6.7%) were thyrotoxic, 1 (0.6%) transient thyrotoxicosis, 1 (0.6%) subclinical hyperthyroidism, 13 (8.0%) had subclinical hypothyroidism, 12 (7.4%) hypothyroidism and 7 (4.3%) had minor changes in thyroid function. CONCLUSIONS: We found a high incidence of new-onset TD, similar to the highest rates reported internationally. Local factors responsible for this need to be investigated. PMID- 15832670 TI - In-hospital outcome of patients discharged from the ICU with tracheostomies. AB - OBJECTIVE: To document the outcome of patients discharged from the intensive care unit (ICU) with tracheostomies. DESIGN AND SETTING: This was a retrospective study conducted in the ICU of Dr George Mukhari Hospital, Pretoria. PATIENTS: All patients discharged from the ICU with tracheostomies over a period of 1 year from 1 January to 31 December 2003. INTERVENTIONS: None. MEASUREMENTS: The main variables studied were post-ICU mortality and length of hospital stay, the Glasgow Coma Scale (GCS) at discharge from ICU and the multiple organ dysfunction score on the day of discharge from the ICU. MAIN RESULTS: Forty-seven patients were discharged with tracheostomies during the study period. The post-ICU mortality was 57%. The mortality of patients discharged with a GCS below 8 was statistically higher than that of patients discharged with a GCS above 8 (79% v. 22%, p = 0.0002). Survivors had significantly longer duration of hospitalisation (26.95 +/- 21.47 days v. 13.48 +/- 14.24 days, p = 0.021) than non-survivors. The mortality rate was higher if the tracheostomy was performed for a low GCS than when it was performed for reasons other than a low GCS (p = 0.0001). The 20 surviving patients were decannulated before discharge from hospital. CONCLUSION: The outcome of patients discharged from the ICU with tracheostomies is, on the whole, unfavourable compared with predicted mortality. A GCS of less than 8 is a good predictor of poor outcome. PMID- 15832671 TI - Electroneurodiagnostics--for the joy of it! AB - During the middle of the last century, discoveries about the brain and the nervous system gave birth to a new technology. Thanks to the pioneers in those efforts, the early EEG technicians, and our efforts through the years, the technology has grown, expanded, and become amazingly sophisticated. This is about loving what we do, and finding joy and satisfaction in our accomplishments. We make health care better. We improve the quality of people's lives. We prompt physicians to sit up and take notice because we complement their efforts at diagnosing, monitoring, and treating people's neurologic illnesses. We're old enough to have a history, and a story...about Kathy Mears and hundreds of dedicated technologists, many of whom sprouted from those first small groups trained by the Fathers of EEG to give them important scientific data. And here we are, fifty-some years later, several thousand strong, influenced by things those first technicians never dreamed about! We're still producing data, but things have changed. Let's examine the challenges electroneurodiagnostic (END) technologists encounter in the current healthcare arena, and focus on what we need to find joy and fulfillment in our important work. Come celebrate our profession. Come share the joy! PMID- 15832672 TI - Review of neonatal EEG. AB - Neonatal electroencephalography (EEG) presents some of the most difficult challenges in EEG interpretation. It differs significantly in many ways from EEG of older children and adults. Technologically, acquisition of a neonatal EEG is significantly more difficult and different than an adult EEG. There are numerous features that are age-specific and change almost week-to-week in the preterm infant. Some features may be normal at one age and abnormal if they persist for several weeks. Many of these features also have different implications in neonates as compared to older individuals. These issues mandate a different approach to neonatal EEG interpretation. In this article an overview of neonatal EEG is presented. After a brief discussion of relevant technical issues, various normal EEG features encountered in neonates are discussed. This is followed by a discussion of the ontogeny of EEG, starting from the age of viability to the first few months of life. A description of various abnormalities follows. Finally, an approach to analysis of a neonatal EEG is presented. PMID- 15832673 TI - Sex-steroid hormones in women with epilepsy. AB - Women with epilepsy face unique challenges, in part related to changes in the seizure threshold at various times in the menstrual cycle or during their reproductive life. Ovarian sex-steroid hormones alter the excitability of the central nervous system (CNS) and alter the frequency and severity of seizures. By understanding the effects of hormones on seizures, clinicians are better equipped to treat hormonally mediated epilepsy. PMID- 15832674 TI - Cortical dysplasia: zones of epileptogenesis. AB - Cortical dysplasia describes a wide range of cerebral cortex structural anomalies and is a condition attributed to multiple etiologies including disruption in the migration of the maturing neurons, disturbance of the process of programmed cell death in the fetal brain, and even noxious environmental influence. Cortical dysplasia can be focal or diffuse and the insult depends more on the timing of the defect in neural development and to a lesser extent the cause. Identification of cortical dysplasia can be done using high resolution magnetic resonance imaging (MRI), histological examination of affected tissue, and EEG. Dysplastic lesions have shown intrinsic epileptogencity. Scalp EEG and electrocorticography (ECoG) reveal several unique patterns including continuous spikes or sharp waves, abrupt runs of high frequency spikes, and periodic spike complexes that occur during sleep. EEG and ECoG can help to guide the surgical resection. Developments in the understanding and treatment of epilepsy caused by cortical dysplasia are occurring rapidly. This article will attempt to provide a brief overview of cortical dysplasia to hopefully prepare and encourage the reader to further investigate cortical dysplasia. PMID- 15832675 TI - National competency skill standards for long-term monitoring in epilepsy. PMID- 15832676 TI - Effect of indole-3-acetic acid on surface properties of the wheat plastid lipids. AB - Surface parameters of polar lipids extracted from winter wheat plastids were investigated by the Langmuir and X-ray differentiation scattering methods. Highly purified plastids were isolated from non-embryogenic (NE) and embryogenic (E) calli initiated from inflorescences. NE plastids contained more monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) and less phospholipids (PL) fraction than E plastids. Moreover, in E calli, unsaturated fatty acids were detected in a higher proportion than in NE for both MGDG and DGDG. No significant differences in fatty acids saturation of PL between NE and E objects were detected. Aqueous surface monolayers were prepared from separate lipids and from mixtures of glycolipids and PL. In the case of MGDG, isotherms showed specific shoulders, contrary to continuous isotherms obtained for other investigated lipids. On the base of pi-A isotherms, the surface parameters: limiting area (A(lim)) and collapse pressure (pi(coll)) were calculated. Indole-3-acetic acid (IAA) increased the A(lim) of all separated lipids about 4-10 angstrom2/mol. However, for NE lipid mixture, the effect of IAA was much smaller (about 2 angstroms2/mol) than for other objects (usually about 5 angstroms2/mol). X-ray experiments for liposomes, obtained from mixtures of glycolipids and PL of NE and E plastids, showed continuous scattering curves with maxima characteristic for lipid bilayer membranes. Calculations of distance distribution functions indicated that bilayer thickness was 41 and 38 angstroms for NE and E, respectively. IAA influence on membrane structures was detected especially in E liposomes and increased the distance between head groups by about 2 angstroms. It is suggested that changes occur during embryogenesis in specific structure of plastid membranes determined also the formation of domains, similar to that suggested for plasmalemma (Plant Sci. 165 (2003) 265). IAA treatment influenced the membrane structure, especially E plastids increasing distances between polar groups. PMID- 15832677 TI - Glycoproteins from sugarcane plants regulate cell polarity of Ustilago scitaminea teliospores. AB - Saccharum officinarum, cv. Mayari, is a variety of sugarcane resistant to smut disease caused by Ustilago scitaminea. Sugarcane naturally produces glycoproteins that accumulate in the parenchymatous cells of stalks. These glycoproteins contain a heterofructan as polysaccharide moiety. The concentration of these glycoproteins clearly increases after inoculation of sugarcane plants with smut teliospores, although major symptoms of disease are not observed. These glycoproteins induce homotypic adhesion and inhibit teliospore germination. When glycoproteins from healthy, non-inoculated plants are fractionated, they inhibit actin capping, which occurs before teliospore germination. However, inoculation of smut teliospores induce glycoprotein fractions that promote teliospore polarity and are different from those obtained from healthy plants. These fractions exhibit arginase activity, which is strongly enhanced in inoculated plants. Arginase from healthy plants binds to cell wall teliospores and it is completely desorpted by sucrose, but only 50% of arginase activity from inoculated plants is desorpted by the disaccharide. The data presented herein are consistent with a model of excess arginase entry into teliospores. Arginase synthesized by sugarcane plants as a response to the experimental infection would increase the synthesis of putrescine, which impedes polarization at concentration values higher than 0.05 mM. However, smut teliospores seem to be able to change the pattern of glycoprotein production by sugarcane, thereby promoting the synthesis of different glycoproteins that activate polarization after binding to their cell wall ligand. PMID- 15832678 TI - The leaf tonoplast V-H(+)-Atpase activity of a C3 halophyte Suaeda salsa is enhanced by salt stress in a Ca-dependent mode. AB - Suaeda salsa seedlings grown in Hoagland nutrient solution were treated with different concentrations of NaCl combined with two levels of Ca2+ (0 and 20 mmol/L) to study the effect of Ca2+ nutrition on the growth and activity of leaf tonoplast V-H(+)-ATPase. Increase of Ca2+ concentration in the solution markedly increased the relative growth quantity of S. salsa seedlings and Ca2+ and K+ concentration in the leaf cell sap under NaCl stress. The leaf V-H(+)-ATPase activity was significantly increased with increasing NaCl concentration under high Ca2+ application (20 mmol/L), but little changed under Ca2+ starvation (0 mmol/L). Western blot analysis showed that the leaf V-H(+)-ATPase of S. salsa was at least composed of A, B, D and c subunits, and their protein amounts were not affected by NaCl treatments under Ca2+ starvation (0 mmol/ L) with an exception of 100 mmol/L NaCl, but increased under high Ca2+ application (20 mmol/L). There was a positive correlation between activity of V-H(+)-ATPase and the protein amounts of the subunits. The results suggest that Ca2+ nutrition played an important role in the salt tolerance of S. salsa, and that enhancement of V-H(+) ATPase activity under salt stress was Ca2(+)-dependent. PMID- 15832679 TI - Alterations in some oxidative parameters in susceptible and resistant wheat plants infected with Puccinia recondita f.sp. tritici. AB - We studied the systemic effects after infection of susceptible and resistant (expressing HSR) wheat plants with leaf rust (Puccinia recondita f.sp. tritici) on the amount of hydrogen peroxide and activity of some ROS scavenging enzymes. Measurements were performed 7 and 21 days after inoculation. In susceptible cultivar (Sadovo 1), an inhibition of activity of catatase and GST was found. By contrast, in resistant cultivar (Kristal), the infection caused an activation of these enzymes. Moreover, it was established that cv. Kristal plants possess constitutive higher levels of hydrogen peroxide, as well as higher superoxide dismutase activity. PMID- 15832680 TI - High temperature effects on photosynthetic activity of two tomato cultivars with different heat susceptibility. AB - The functional activities of the photosynthetic apparatus of two tomato cultivars of different thermotolerance were investigated after a short period of high temperature treatment. Seedlings of two tomato genotypes, Lycopersicon esculentum var. Campbell-28 and the wild thermotolerant Nagcarlang, were grown under a photoperiod of 16h at 25 degrees C and dark period of 8h at 20 degrees C. At the fourth true leaf stage, a group of plants was exposed to heat stress of 45 degrees C for 2 h. The heat shock treatment caused important reductions of the net photosynthetic rate (Pn) of Campbell-28 plants due to non-stomatal components. These non-stomatal effects were not evident in Nagcarlang-treated plants. This reduction in the CO2 assimilation rate observed in Campbell-28 was generated by affections in the Calvin cycle and also in the PSII functioning. No changes in these parameters were observed in the thermotolerant genotype after the stress. Injury to the plasma membrane because of the heat stress was evident only in the Campbell-28 genotype. Heat led to a sun-type adaptation response of the photosynthesis pigment apparatus for the Nagcarlang genotype, but not for Campbell-28, and thus an increase in chlorophyll a/b ratio and a decrease in chlorophyll/carotenoid ratio were shown in Nagcarlang stressed plants. PMID- 15832681 TI - Expression of ascorbate peroxidase and glutathione reductase in roots of rice seedlings in response to NaCl and H2O2. AB - The accumulation of H2O2 by NaCl was observed in the roots of rice seedlings. Treatment with NaCl caused an increase in the activities of ascorbate peroxidase (APX) and glutathione reductase (GR) and the expression of OsAPX and OsGR in rice roots. Exogenously applied H2O2 also enhanced the activities of APX and GR and the expression of OsAPX and OsGR in rice roots. The accumulation of H2O2 in rice roots in response to NaCl was inhibited by the NADPH oxidase inhibitors, diphenyleneiodonium chloride (DPI) and imidazole (IMD). However, DPI, IMD, and dimethylthiourea, a H2O2 trap, did not reduce NaCl-enhanced activities of APX and GR and expression of OsAPX and OsGR. It appears that H2O2 is not involved in the regulation of NaCl-induced APX and GR activities and OsAPX and OsGR expression in rice roots. PMID- 15832683 TI - Nodule conductance varied among common bean (Phaseolus vulgaris) genotypes under phosphorus deficiency. AB - Common bean genotypes BAT477, COCOT, DOR364, Flamingo, and NAG310 were inoculated with Rhizobium tropici CIAT899 and grown under phosphorus deficiency. This treatment induced a significant decrease in shoot and nodule growth that varied among genotypes from 35% to 57% and from 45% to 61%, respectively, whereas root biomass was less affected. Phosphorus deficiency affected differently the genotypes for nodule number and size, and the responses of nodulated-root O2 uptake (Conr) to raising rhizospheric PO2. From the later data, nodule conductance could be computed by dividing the slope of the regression of Conr as a function of external pO2 by nodule surface area. It is concluded that differences in nodule conductance are related to genotypic tolerance to P deficiency. PMID- 15832682 TI - Possible role of catalase in post-dormancy bud break in grapevines. AB - Changes in the activity of catalase (Cat) and in the levels of H2O2 were followed throughout dormancy in buds of grapevines (Vitis vinifera L.). In grapevines grown in the Elqui valley in Chile, a region with warm-winters, the activity of Cat increased during the recess period of buds, reaching a maximum and thereafter decreased to less than one third of its maximal activity. Three isoforms of Cat were detected in extracts of buds by native PAGE analysis, and the extracted activity was inhibited competitively by hydrogen cyanamide (HC), a potent bud break agent. Furthermore, HC applications to field-grown grapevines in addition to the expected effect on advancing bud break, reduced the Cat activity during bud dormancy. Similar reductions were observed during dormancy in buds of grapevines grown in the Central valley in Chile, a region with temperate winters, suggesting that HC and winter chilling inhibits the activity of the main H2O2 degrading enzyme in grape buds. A transient rise in H2O2 levels preceded the release of buds from endodormancy, moreover, the peak of H2O2 and the onset of bud break occurred earlier in HC treated than in control grapevines, suggesting the participation of H2O2 as a signal molecule in the release of endodormancy in grape buds. The relationship between Cat inhibition, rise in H2O2 levels and initiation of bud break are discussed. PMID- 15832684 TI - Effects of nitric oxide on growth of maize seedling leaves in the presence or absence of ultraviolet-B radiation. AB - The leaves of maize seedlings were used to measure leaf biomass including leaf length, width and weight, and to examine the relationship between nitric oxide (NO) synthase activity in microsomes and cytosol to the exo- and endo-beta glucanase activity during growth. It was found that ultraviolet-B radiation (UV-B radiation) strongly induced nitric oxide synthase (NOS) activity but caused both a decrease of leaf biomass and exo- or endo-beta-glucanase activity. In contrast, the NOS inhibitor and NO donor largely decreased the activity of NOS in non irradiated seedlings. The inhibitor also reduced exo- and endo-beta-glucanase activity and leaf biomass while the donor increased the enzyme activity and leaf biomass under normal conditions. Alternatively, under ultraviolet-B, the additional inhibitor of NOS and NO donor appeared to compromise the effects of ultraviolet-B on glucanase activity and leaf biomass, making the relationship between NOS activity and glucanase activity negatively correlated. This suggests that the changes of NOS activity showed a positive correlation to glucanase activity and leaf biomass in the absence of ultraviolet-B, but a negative correlation to ultraviolet-B irradiation and NO donor treatment alone. It is assumed that exo- and endogenous NO is responsible for the up-regulation of regular growth and development without ultraviolet-B. Under UV-B radiation, however, it might function as a signaling molecule of ultraviolet-B inhibiting leaf growth of maize seedlings to carry out stress-signaling transduction. PMID- 15832685 TI - Relationship between xylem ion concentration and bean growth responses to short term salinisation in spring and summer. AB - Bean plants, Phaseolus vulgaris L. cv. Contender, were grown in the spring and summer seasons to study the relationship between xylem Na+/Cl-, transpiration rate, and salt tolerance. Eight-day-old seedlings were transplanted to 50% modified Hoagland solution with 1 mM NaCl. Four days after transfer, one of two treatments was applied: a control of 1 mM NaCl or a treatment of 25 mM NaCl every two days to reach a final treatment concentration of 75 mM NaCl. Plants were sampled on the fourth day after the final salt concentration was reached, eight days after the salinisation treatment began. Relative growth rate was 2.6-fold greater in summer than in spring. However, while no differences were found between treatments in spring, summer salt-treated plants had growth rates that were 31% lower than those of controls. In summer, CO2 assimilation, stomatal conductance, and transpiration rate of salinised plants declined with respect to controls. Leaf Na+ and trifoliolate leaf Cl- were higher in salt-treated plants in summer, although root Na+ was significantly higher in spring. Moreover, in summer salinity inhibited Ca2+ and K+ uptake and changed its distribution. Summer salt-treated plants had an average of 17-fold higher xylem Na+ during the daily cycle, while xylem Cl-, only in the afternoon, showed higher values (1.5-fold) compared to spring-grown plants. Our results suggest that the faster growth response to salt in summer-grown bean was at least partly due to an increase in xylem Na+ independent of the transpiration rate and possibly related to an increase in xylem Na+ influx or/and Na+ recirculation. PMID- 15832686 TI - Physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron deficiency. AB - Plants are frequently submitted to iron deficiency when growing on calcareous soils, which contain high concentrations of bicarbonate. The purpose of this study was to investigate the variability of physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron chlorosis. Vine woodcuttings of seven autochthonous Tunisian varieties (Khamri, Mahdaoui, Blan3, Saouadi, Arich Dresse, Beldi and Balta4), two rootstocks (140Ru and S.O.4), and an introduced table variety (Cardinal) were cultivated on inert sand for 2 months using a complete nutrient solution (20 microM Fe) that was either well supplied or not supplied with 10 mM HCO3-. Young leaves of plants cultivated on bicarbonate enriched medium showed characteristic symptoms of iron chlorosis, although the intensity of the symptoms depended on the variety and the rootstock. Chlorosis score confirmed these observations since the most sensitive varieties showed the highest values. This variability in tolerance to iron deficiency was also displayed when analysing the physiological parameters (shoot length, plant dry weight, and chlorophyll concentration) and the iron contents in the 4th leaf. Analysis of morphological and physiological parameters showed three behaviour groups. The first one corresponded to tolerant varieties (Khamri, Mahdaoui, and the root-stock: 140Ru), the second included moderately tolerant vines (Saouadi, Arich Dresse, Blanc3, and the rootstock: S.O.4) and the third represented the sensitive ones (Balta4, Beldi, and Cardinal). PMID- 15832687 TI - Interactive effects of phosphate deficiency, sucrose and light/dark conditions on gene expression of UDP-glucose pyrophosphorylase in Arabidopsis. AB - The effects of inorganic phosphate (Pi) status, light/dark and sucrose on expression of UDP-glucose pyrophosphorylase (UGPase) gene (Ugp), which is involved in sucrose/ polysaccharides metabolism, were investigated using Arabidopsis wild-type (wt) plants and mutants impaired in Pi and carbohydrate status. Generally, P-deficiency resulted in increased Ugp expression and enhanced UGPase activity and protein content, as found for wt plants grown on P-deficient and complete nutrient solution, as well as for pho1 (P-deficient) mutants. Ugp was highly expressed in darkened leaves of pho1, but not wt plants; daily light exposure enhanced Ugp expression both in wt and pho mutants. The pho1 and pho2 (Pi-accumulating) mutations had little or no effect on leaf contents of glucose and fructose, regardless of light/dark conditions, whereas pho1 plants had much higher levels of sucrose and starch in the dark than pho2 and wt plants. The Ugp was up-regulated when leaves were fed with sucrose in wt plants, but the expression in pho2 background was much less sensitive to sucrose supply than in wt and pho1 plants. Expression of Ugp in pgm1 and sex1 mutants (impaired in starch/sugar content) was not dependent on starch content, and not tightly correlated with soluble sugar status. Okadaic acid (OKA) effectively blocked the P-starvation and sucrose-dependent expression of Ugp in excised leaves, whereas staurosporine (STA) had only a small effect on both processes (especially in -P leaves), suggesting that P-starvation and sucrose effects on Ugp are transmitted by pathways that may share similar components with respect to their (in) sensitivity to OKA and STA. The results of this study suggest that Ugp expression is modulated by an interaction of signals derived from P-deficiency status, sucrose content and dark/light conditions, and that light/sucrose and P deficiency may have additive effects on Ugp expression. PMID- 15832688 TI - Hyperhydricity in shoot cultures of Scrophularia yoshimurae can be effectively reduced by ventilation of culture vessels. AB - An effective procedure for obtaining healthy shoots from nodal segments of Scrophularia yoshimurae is described. Nodal segments cultured on Murashige and Skoog's (MS) basal medium supplemented with 1.0 mg L(-1) benzyladenine (BA) and 0.2 mg L(-1) alpha-naphthaleneacetic acid (NAA) induced multiple shoots in conical flasks that differed in the way they were closed and sealed. Hermitically sealed culture vessels resulted in high hyperhydricity/vitrification. High concentrations of ethylene and CO2 were found to accumulate in these vessels. The hyperhydricity of the shoot cultures could be decreased by progressively ventilating the vessels. Exchange of gases was achieved by removing the Parafilm sealing without affecting sterility. This reduced the hyperhydricity rate and gave a good recovery of vitrified shoots, but resulted in decreased proliferation and a dehydration of proliferating nodal segments and the culture medium. The best number of normal shoots was observed when the parafilm was removed for gaseous exchange after four weeks of culture incubation. The results show that hyperhydricity in shoot cultures of S. yoshimurae could be prevented by sufficient gas exchange during culture. PMID- 15832689 TI - A common factors approach to improving mental health services. PMID- 15832690 TI - Identifying and selecting the common elements of evidence based interventions: a distillation and matching model. AB - A model is proposed whereby the intervention literature can be empirically factored or distilled to derive profiles from evidence-based approaches. The profiles can then be matched to individual clients based on consideration of their target problems, as well as demographic and contextual factors. Application of the model is illustrated by an analysis of the youth treatment literature. Benefits of the model include its potential to facilitate improved understanding of similarities and differences among treatments, to guide treatment selection and matching to clients, to address gaps in the literature, and to point to possibilities for new interventions based on the current research base. PMID- 15832691 TI - Client expectancies about therapy. AB - Client expectancies about therapy are considered a common factor, that is, a feature of therapy that is not specific to any particular technique or approach. This review includes both child- and adult-focused studies from the expectancies literature, emphasizing findings about the relationship between expectancies and factors including client improvement, attrition, and therapeutic alliance (TA). Results suggest that expectancies are related to both client improvement and TA, but do not support a relationship between expectancies and attrition. We then present two preliminary hypothesized pathways of how pretreatment client characteristics and therapy outcomes relate to role and outcome expectancies. Implications of these pathways are discussed. PMID- 15832692 TI - A theoretical model of common process factors in youth and family therapy. AB - Recently there has been an increasing emphasis in the youth and family mental health treatment literature on the use of empirically supported treatments (ESTs). In contrast there has been scant attention paid to more universal aspects of the therapy process that may have even greater impact upon therapy outcomes. It is likely that the success of the techniques proposed by ESTs may depend on the presence of common process factors. In this article, the authors explore the status of common process factors research in the youth and family therapy literature, and propose a theoretical model linking specific therapeutic relationship variables and treatment outcomes for children and adolescents. This model is intended to guide synthesis of the empirical evidence for common process factors in youth and family treatment and to stimulate future research on common process factors. PMID- 15832693 TI - What is the evidence for evidence-based treatments? A hard look at our soft underbelly. AB - In the rising quest for evidence-based interventions, recent research often does not give adequate attention to "nonspecific therapeutic factors," including the effects of attention, positive regard, and therapeutic alliance, as well as the effects of treatment dose, intensity and actual processes mediating therapeutic change. To determine the extent to which recent clinical trial designs fully this problem, the authors conducted a systematic review of Psych-Lit/Medline of all controlled child psychotherapy treatment studies from 1995 to 2004. A total of 52 studies were identified that met review criteria: two or more therapy conditions and random assignment of participants to intervention groups. Of the 52 studies, one group (n = 29) compared a presumably active treatment with 1 or more similarly intensive treatments (often an attention control group) presumably not containing the active therapeutic ingredients. Of these, 14 studies found evidence of consistent differences between the two groups, whereas 15 did not. An additional group of studies (n = 27) compared therapy groups with different levels of intensity and "dose" of the putatively active treatment; 13 of these found evidence of the effects of different levels of treatment dose/intensity on outcomes and 14 did not. Four studies met criteria for inclusion in both groups. Across both groups of studies, when positive effects were found, few studies systematically explored whether the presumed active therapeutic ingredients actually accounted for the degree of change, nor did they often address plausible alternative explanations, such as nonspecific therapeutic factors of positive expectancies, therapeutic alliance, or attention. Findings suggest that many child psychotherapy treatment studies have not inadequately controlled for nonspecific factors such as attention and treatment intensity and have failed to assess specific mediators of change. Specific recommendations for future studies are offered, specifically: 1. Initial specification in study design how investigators will test if the intervention is efficacious over and above the effects of positive expectancies, positive regard, or attention; 2. Planned (rather than post hoc) analyses to explore whether and how a given treatment's specific ingredients or the overall intensity of its "dose" are related to treatment outcomes; 3. More studies conducting head-to-head tests of different types but equally credible forms of treatment, with planned analyses testing different mechanisms of change 4. When positive effects of a treatment vs. a control are found, systematic elimination of specific, competing hypotheses of reasons for treatment efficacy (e.g., attention, therapeutic alliance, face validity of treatment and client's treatment expectations, changes in self efficacy), and 5. When negative effects are reported, appropriate analyses to address alternative explanations (lack of power, floor or ceiling effects, mediator analyses to address possible fidelity or adherence problems, therapist effects, absence of main effects but possible subgroup/moderator effects, etc.). PMID- 15832694 TI - [Management of medically unexplained symptoms]. PMID- 15832695 TI - [Just a hernia--for the last time?]. PMID- 15832696 TI - [General practice at the universities (GPU). Danish Society of General Practice]. PMID- 15832697 TI - [Acupuncture and research. Danish Medical Society of Acupuncture]. PMID- 15832698 TI - [Activity-based registration of computer use in epidemiological studies. Danish Society of Occupational and Environmental Medicine]. PMID- 15832699 TI - [Deep brain stimulation of therapy-refractory, disabling dystonia. Danish Society of Movement Disorders (Danmodis)]. PMID- 15832700 TI - [Attention deficit hyperactivity disorder. Danish Society of Child and Adolescent Psychiatry in Denmark]. PMID- 15832701 TI - [Overweight among school children in Ishoj municipality. Danish Society of Child Health]. PMID- 15832702 TI - [Diabetes and heart disease. Danish Society of Cardiology]. PMID- 15832703 TI - [Diagnostic imaging of skin diseases. Danish Society of Dermatology]. PMID- 15832704 TI - [Treatment of epilepsy--where are we now? Danish Society of Epilepsy]. PMID- 15832705 TI - [Clinical pharmacology: important events in 2004. Danish Society of Clinical Pharmacology]. PMID- 15832706 TI - [Magnetic resonance imaging of the heart. Danish Society of Clinical Physiology and Nuclear Medicine]. PMID- 15832707 TI - [Is treatment of old people beneficial? Danish Society of Geriatrics]. PMID- 15832708 TI - [Good clinical practice. Danish Society of Good Clinical Practice]. PMID- 15832709 TI - [Greenland--a changing disease pattern. Greenland Medical Society]. PMID- 15832710 TI - [Functional imaging of the liver with positron emission tomography. Danish Society of Hepatology]. PMID- 15832711 TI - [Antibodies combined with radiotherapy in the curative treatment of head and neck cancer. Danish Society of Head-Neck Oncology and Danish Head-Neck Cancer Group]. PMID- 15832712 TI - [Chronic lymphatic leukemia: primary treatment based on biological risk stratification. Danish Society of Hematology]. PMID- 15832713 TI - [Avian influenza. Danish Society of Infection Medicine]. PMID- 15832714 TI - [Surgical simulation and organization. Danish Society of Surgery]. PMID- 15832715 TI - [Screening and prevention. Danish Society of Pulmonary Medicine]. PMID- 15832716 TI - [Epidemic increase of methicillin resistant Staphylococcus aureus (MRSA) in Denmark--interventions required now]. PMID- 15832717 TI - [Spinal surgery organization. Danish Society of Neurosurgery]. PMID- 15832718 TI - [Thrombolytic therapy of acute ischemic apoplexy. Danish Society of Neurology]. PMID- 15832719 TI - [A new offer to all pregnant women: screening for Down syndrome and congenital abnormalities. Danish Society of Obstetrics and Gynecology]. PMID- 15832721 TI - [Pathology database. Danish Society of Pathologic Anatomy and Cytology]. PMID- 15832720 TI - [DiaBase--a national clinical quality database for systematic screening of diabetic retinopathy. Danish Society of Ophthalmology]. PMID- 15832722 TI - [Psychiatry--prevention and specializing. Danish Society of Psychiatry]. PMID- 15832723 TI - [Combined urinary and fecal incontinence--a neglected problem. Danish Society of Pediatrics]. PMID- 15832724 TI - [Liver ultrasonography with the use of contrast media. Danish Society of Radiology]. PMID- 15832725 TI - [Multiple sclerosis. Danish Society of Multiple Sclerosis Research (DAREMUS)]. PMID- 15832726 TI - [While waiting for the Danish Quality model. Danish Society of Quality in Health Care (DSKS)]. PMID- 15832727 TI - [Interdisciplinary education and knowledge of wound management. Danish Society of Wound Healing]. PMID- 15832728 TI - [It can be beneficial! Danish Society of Tropical Medicine and International Health]. PMID- 15832729 TI - [Educational strategies for diagnostic ultrasonography. Danish Society of Diagnostic Ultrasonography]. PMID- 15832730 TI - [Organization of urology. Danish Society of Urology]. PMID- 15832731 TI - [Creams used in the treatment of children can cause cancer]. PMID- 15832732 TI - [Gastrointestinal stromal tumours of the stomach: personal experience]. AB - Gastrointestinal stromal tumours are a relatively new nosological entity with an increasing incidence. CD117 positivity at immunohistochemical analysis is mandatory to distinguish them from other mesenchymal tumours. A gastric localisation is observed in more than 70% of cases. In the present paper 11 primary gastrointestinal stromal tumours of the stomach operated during the last decade at our institution were retrospectively reviewed. Hospital files and histological slides of 11 patients with gastrointestinal stromal tumours of the stomach undergoing surgical resection at our institution over the period from 1993 to 2003 were reviewed retrospectively. The variables analysed were: morphological and immuno-histochemical characteristics of the tumours, demographic data, type of surgical treatment and postoperative course. Long-term survival was evaluated on the basis of clinical and/or telephone follow-up in all patients. Immunohistochemical analysis for CD11 7 proved positive in all patients. The size of the tumours ranged from 3 to 23 cm: tumour size was > 5 cm and > 10 cm in 8 and 3 patients, respectively. Nodal involvement was detected in one patient and another had liver metastases. Surgical treatment consisted of wedge resection in 3 cases, distal subtotal gastrectomy in 4 and total gastrectomy in 4. In 2 patients surgical excision was extended to other organs. No postoperative mortality or major postoperative complications were observed. Nine patients were still alive at follow-up; 1 patient died as a result of a neoplastic relapse (38 months) and 1 died of other causes (48 months). The median survival was 42 months. Gastrointestinal stromal tumours are characterised by slow growth and therefore clinical signs are delayed. For that reason large tumours are often observed at the time of diagnosis. Surgical resection is the only potential curative treatment; but the risk of recurrence (local or at distance) remains high. PMID- 15832733 TI - [Preliminary results of neoadjuvant treatment of adenocarcinoma of the gastro esophageal junction]. AB - The prognosis of adenocarcinoma of the gastro-oesophageal junction is poor and only surgery yields long-term survival in no more than 30% of patients. We tested a new neoadjuvant chemo-radiotherapy regimen based on the administration of weekly docetaxel and cisplatin and continuous infusion of 5-FU with concurrent radiotherapy in order to evaluate its feasibility and efficacy. Thirty-three patients enrolled in a dose-finding study and observed at the 1st Division of General Surgery of the University of Verona between January 2000 and October 2003 underwent neoadjuvant chemo-radiotherapy for gastro-oesophageal junction adenocarcinoma (Siewert type I and II). The induction treatment was completed in 97.0% of cases with no treatment-related mortality. After completion of chemo radiation 30 patients underwent surgery (90.9%) while three patients did not (progression in 2 cases and chemotherapy toxicity in one). Two operated patients did not undergo resection because of liver metastasis at laparotomy (respectability: 84.8%) and 3 more cases had incomplete tumour resection (R0 resectability: 75.8%). No postoperative in-hospital mortality was observed. A complete response (pT0N0) was achieved in 7 cases (23.3%) while minimal residual disease without evidence of lymph node involvement was found in a further 5 cases (16.7%). Worthy of note is the high rate of positive histopathological responses in the later period (6 out of 8) with 4 cases presenting complete responses. This protocol regimen proved to be feasible and well tolerated. Surgery-related deaths and morbidity were not increased. A high rate of positive pathological responses was obtained particularly in the later period of the study with the increased dosage of the protocol regimen. PMID- 15832734 TI - [Laparoscopic liver resection without a Pringle maneuver for HCC in cirrhotic patients]. AB - In the belief that the advantages stemming from a minimally invasive approach are significant, particularly in cirrhosis patients, we decided to apply this technique in the treatment of a group of patients suffering from HCC associated with cirrhosis. Sixteen patients (10 men, 6 women; mean age 60.1 years) underwent laparoscopic surgery for HCC associated with well compensated HCV-related liver cirrhosis (Child-Pugh class A; mean tumour size 2.9 cm). Seven of these lesions were located in the left liver and 9 in the right lobe. Laparoscopy was performed with a CO2 pneumoperitoneum (12-14 mmHg). The Pringle manoeuvre was not used. There was one conversion to laparotomy due to inadequate exposure. We performed 13 non-anatomical resections, 1 VI segmentectomy and 1 anatomical left lobectomy. None of the patients required blood transfusions. One patient died of severe respiratory distress syndrome on postoperative day 3. Major morbidity included 2 moderate postoperative ascites successfully resolved with conservative treatment. To date (mean follow-up: 18 months) no recurrences at the resection site or port site metastases have been observed. Limited laparoscopic liver resections for HCC in cirrhotic patients are technically feasible and safe when careful selection criteria are adopted (hepatic involvement limited and located in the left or anterior right segments, tumour size smaller than 5 cm, Child-Pugh class A). PMID- 15832735 TI - [Does the Roux-en-Y-stasis syndrome still exist?]. AB - Roux-en-Y-stasis syndrome (RYS) is a complication of subtotal gastrectomy characterized by delayed gastric emptying and vomiting. The aim of the study was to analyze RYS frequency with particular attention to diagnosis and therapy. From November 1996 to June 2004, we performed 147 distal gastrectomies with 5 cases of RYS: mean age 78 years, 3 male, 2 female, 4 adenocarcinoma and 1 GIST. Among the 5 cases, RYS was due to different causes: it was functional in 2 cases (with difficult gastric emptying due to a long gastric remnant in one patient, while the other was associated with duodenal fistula) and healed through medical therapy in both; a third patient had an edematous stenosis of the gastrojejunal anastomosis treated with medical therapy and the remaining 2 patients had jejunal obstruction due to adherences and required reoperations. RYS is a rare complication of subtotal gastrectomy determined by different causes. Recognizing the cause is very important for choice of appropriate therapy. PMID- 15832736 TI - [Therapy of asymptomatic gallstones: indications and limits]. AB - Gallstone disease is one of the most common health problems world-wide. It is also one of the main causes of medical expenditure in Western countries. Asymptomatic gallstones are defined as stones that have not given rise to biliary cholic or other biliary symptoms. A number of epidemiological cross-sectional screening studies have shown that as many as 66 to 77% of patients with gallstones are asymptomatic. Opinion regarding the development of the disease in the asymptomatic patients has changed in the course of time. In 1992 Friedman, in his review of literature, established that only 1-2% of asymptomatic patients developed severe symptoms or complications early, with fewer complications developing in later years than in the years soon after discovery of the gallstones. Recent prospective epidemiological studies have established that no particular factor has any effect on the natural course of the disease. Laparoscopic cholecystectomy is the gold standard for symptomatic gallstones, but the management of patients with asymptomatic stones remains controversial. This problem is related to the incidence of biliary lesions which has remained substantially unchanged over the past few years. It has recently been demonstrated that the risk of iatrogenic lesions is not entirely dependent upon the surgeon's experience. There is a substantial consensus of agreement that surgical treatment has an unfavourable cost:benefit ratio in asymptomatic patients. The Authors conclude that, because of the condition's benign natural history, a wait-and-see policy is to be recommended in all asymptomatic patients, except for only a few selected cases. PMID- 15832737 TI - [Inguinal hernia recurrence. Report of our personal experience]. AB - In our previous studies, we reasserted the concept regarding the fundamental role of the transversalis fascia in securing and maintaining restraint on the abdominal wall in correspondence with the "inguinal fissure", which is considered a hiatus and a weak point. The role played by the transversalis fascia was first proposed by Edoardo Bassini and later reaffirmed by other authors, surgeons and anatomists. This concept led to the belief that the procedures deemed effective in treating inguinal hernia were those capable of strengthening or substituting the transversalis fascia. In a recent study we compared the immediate and long term results of three techniques: Bassini, Lichtenstein and preperitoneal prosthesis by inguinal way. A lower number of recurrences was noted in the latter as compared to the others. Based on the aforementioned points, the same verification was carried out regarding recurrent inguinal hernia. The results concerning the incidence of relapse were similar, proving once again the effectiveness of the preperitoneal prosthesis (2.5% vs 6.6% for Lichtenstein). We have also attempted to identify the factors which were considered possible causes of recurrence in the previous cases we examined, allowing us to reach useful conclusions in the prevention of hernia recurrence. PMID- 15832738 TI - [Laparoscopic Nissen fundoplication and esophageal intestinal metaplasia: preliminary observations]. AB - The aim of this preliminary study conducted in a few cases was the retrospective evaluation of the effects of laparoscopic Nissen fundoplication on oesophageal intestinal metaplasia. Seventy-seven patients with hiatal hernia underwent digital videofluorography, endoscopy with biopsies, motility studies and 24-h oesophageal pH-monitoring. On the basis of the results of the diagnostic procedures and considering the patients' ages and response to proton-pump inhibitor treatment, 8 patients underwent laparoscopic Nissen fundoplication; in 5 cases intestinal metaplasia was present at histopathological examination. Two of these had Barrett's oesophagus at endoscopy and intestinal metaplasia was associated with low-grade dysplasia in both at histology; the other 3 did not present a columnar mucosa at endoscopy and 1 had low-grade dysplasia. In all 5 patients, at 1 year postoperative histopathological control, disappearance or decrease of metaplastic epithelium and regression of dysplasia were noted, with excellent results in terms of reflux symptoms at clinical control. On the basis of these preliminary data, it is our opinion that antireflux surgery is not only a suitable treatment in the management of Barrett's oesophagus but also has a favourable effect on intestinal metaplasia regression when a normal oesophageal mucosa is present. PMID- 15832739 TI - [Digital videofluorography and esophageal achalasia: from diagnosis to follow up]. AB - The aim of this retrospective study was to evaluate the use of digital videofluorography in the preoperative and postoperative management of esophageal achalasia surgical treatment. From 1990 to 2004, 25 patients with achalasia, diagnosed by digital videofluorography and confirmed by motility studies and endoscopy, underwent surgery. All patients underwent digital videofluorography at 1, 6 and 12 months in order to evaluate the completeness of the myotomy and the efficacy of the antireflux procedures. At postoperative videofluorography esophageal transit time was decreased in all patients (100%); esophageal motor activity was unchanged in 23 (92%), and modified in two patients (8%) with onset of peristaltic-like motor activity; 8 patients (35%) presented decreased preoperative dilatation; all patients had a WST negative for post-myotomy reflux. On the basis of our experience and the advantages of the procedure we suggest videofluorography as a first-approach diagnostic examination useful for surgical indications and postoperative follow-up in achalasic patients. PMID- 15832740 TI - [The role of fine-needle aspiration in the diagnosis of thyroid nodules]. AB - The aim of this study was to prospectively evaluate the sensitivity, specificity and accuracy of fine-needle aspiration of thyroid nodules. From January 1978 to December 2003, 497 patients underwent fine-needle aspiration of thyroid nodules and then thyroidectomy. Fine-needle aspiration was performed with a fine 23-25 gauge and the aspirate was fixed and dyed with May-Grounwald-Giemsa method. The results of cytology were divided into four groups: positive for malignancy (77 patients), negative for malignancy (370 patients), suspect for malignancy (34 patients) and not diagnostic (16 patients). Suspect specimens also included follicular neoplasm and Hurthle cell neoplasm. Fine-needle aspiration results were compared with histopathologic analysis after thyroidectomy. There were 2 false-positive (0.5%) and 5 false-negative patients (1%). Sensitivity, specificity and accuracy were respectively 94.7%, 99% and 98.4%. This study confirms that fine-needle aspiration of thyroid nodules can be performed easily with high sensitivity, specificity and accuracy. PMID- 15832741 TI - [Blunt liver trauma: therapeutic options]. AB - Therapeutic options for management of blunt hepatic trauma include both non operative and operative management, until liver transplantation. The presence of associated intraabdominal lesions increases mobility and mortality. A total of 58 patients with blunt hepatic injury, observed at the 2nd Surgical Unit of the "Santa Chiara Hospital" of Trento over a four-year period (1999-2003), were analyzed. Thirty-eight patients (65.5%) were treated conservatively with a 94.7% success rate. Twenty (34.5%) patients were surgically treated. Suture hepatorraphy was the most common procedure performed. The overall morbidity in surgical patients was 25%, with 20% of liver related complications. Four patients (20%) died with a liver related mortality of 10%. Advantages of non-operative management included a reduced need for transfusion (1.1 vs. 4.3 U), with 81.5% of patients not requiring transfusion, a reduced stay in the ICU and a reduced rate of morbidity and mortality (2.6% vs. 20%). Hemodynamic instability and the presence of an associated lesion in blunt hepatic trauma patients require emergency laparotomy. The widespread application of non-operative management has reduced the rate of morbidity and mortality related to unnecessary surgical approaches. PMID- 15832742 TI - [Efficacy of topical use of 0.2% glyceryl trinitrate in reducing post haemorrhoidectomy pain and improving wound healing]. AB - The aim of the study was to evaluate whether topical application of 0.2% glyceryl trinitrate ointment could reduce post-haemorrhoidectomy healing time and pain both at rest and during defecation. Thirty patients with grade III and IV haemorrhoids were included in the study and divided into two groups. All patients underwent Milligan-Morgan haemorrhoidectomy, and anorectal manometry was performed before surgery and after 5 and 30 days. In one group a placebo ointment was applied to the perianal wounds, while in the other group a 0.2% glyceryl trinitrate ointment was used. Maximum resting pressure was reduced in the glyceryl trinitrate group and increased in the placebo group after 5 days. Postoperative pain both at rest and during defecation, and the time to healing and return to normal activity were significantly reduced in the glyceryl trinitrate group, whilst analgesic consumption was similar. An elevated incidence of headache was observed In the glyceryl trinitrate group. Topical application of glyceryl trinitrate was effective in reducing postoperative pain and healing time, but the substantial incidence of side effects may limit its extensive use. PMID- 15832743 TI - [Trocars and hernias: a simple, cheap remedy]. AB - Trocar-site incisional hernias and their complications are reported in 1% to 6% of patients. Such hernias are attributed to the difficulty of applying standard suturing techniques to wound closure. We report our experience with a simple device, the Deschamps ligature needle. PMID- 15832744 TI - [Report on a case of schwannoma of the neck: clinical and therapeutic considerations]. AB - We present a case of schwannoma of the neck in 49-year-old man, which was symptomatic for a long period: paraesthesia for one year, cervical mass for 2-3 months and dysphagia during the last 20 days. After ultrasonography and magnetic resonance the patient was operated on and complete excision of the mass was performed. The operation was performed via a cervical approach: the nerves and vascular and muscular structures were carefully isolated and then preserved. The tumour arose from the cervical sympathetic chain. The diagnosis of schwannoma was possible only at histopathological examination. After 24 months no local recurrence or postoperative complications related to the intervention were found. At present, however, the patient presents a moderate lowering of the voice, such as may be due to impairment of the superior laryngeal nerve, though this was already present preoperatively. These lesions are uncommon and most often occur as asymptomatic solitary neck masses. The preoperative diagnosis may be difficult, even with the aid of ultrasound, computed tomography, magnetic resonance imaging, and angiography. Identification of the nerve is often difficult up to the time of surgery, which is the treatment of choice for schwannoma. PMID- 15832745 TI - [Composite tumour of the stomach: a case report and review of the literature]. AB - The authors report a rare case of composite tumour of the stomach in a 53-year old woman documenting the neuroendocrine and glandular features of the tumour by means of immunohistochemical investigations. The authors examine the diagnostic and therapeutic possibilities currently available for the management of this group of rare tumours of uncertain histogenesis. They conclude in favour of elective surgical treatment depending on the site and stage of the tumour in keeping with the data reported in the literature. They also discuss the prognosis of such tumours. PMID- 15832746 TI - Tumoral calcinosis: a case report. AB - Tumoral calcinosis is a rare tumour-like mass characterized by soft tissue calcification of obscure aetiology. A case of tumoral calcinosis is presented here, and its clinical, radiological and pathological features are described. The differential diagnosis versus hydatid cyst is discussed. Diagnosis is possible with imaging techniques but histopathological study is essential to establish it with certainty. Complete surgical excision appears to be the only effective treatment. PMID- 15832747 TI - [Traumatic rupture of the pancreatic isthmus complicated by concomitant rupture of the duodenum and right kidney]. AB - The Authors describe a case of complete traumatic transection of the pancreatic isthmus associated with complete transection of the first portion of the duodenum and the isthmus of a horseshoe kidney. The treatment of the pancreatic lesion was intestinal drainage of the distal portion of the pancreas and closure of the proximal one. The authors stress the advantages and good outcome of the conservative operation, performed at an early stage, with the clinical indications and adequate surgical techniques. PMID- 15832748 TI - [Neuroendocrine carcinoma of the colon with metastasis to the brain: a case report and review of the literature]. AB - The Authors report a case of a 71-year-old male with synchronous neuroendocrine colon carcinoma and a solitary brain metastasis. The patient was treated surgically with resection of both the cerebral and intestinal lesions followed by cerebral radiotherapy. A pulmonary metastasis was discovered after 3 months and treated with interferon and octreotide. No further cerebral or intestinal relapses were observed. The patient died of cardiac disease 11 months after the first operation. Central nervous system metastases from carcinoid tumours are rare. The reported survival in untreated cases or in cases treated by radiotherapy alone is 4 months. In the case reported, surgical resection of the cerebral and intestinal lesions followed by intracranial radiotherapy yielded complete local control of the disease and a slight improvement in survival. PMID- 15832750 TI - [New orientations in the management of advanced, metastatic gastrointestinal stromal tumors (GIST): combination of surgery and systemic therapy with imatinib in a case of primary gastric location]. AB - Gastrointestinal stromal tumours (GIST) are rare neoplasms originating from connective tissue in the digestive tract with an incidence of less than 1% and account for most non-epithelial primitive digestive tumours. Metastasis diagnosed at the time of disease discovery confirms GIST malignancy. Kit protein, a trans membrane tyrosine kinase receptor of staminal cells, is characteristically expressed by GIST. Most GIST have a mutation in the kit proto-oncogene. Resistance to conventional chemotherapy is commonly shown by malignant GIST. Most patients with advanced malignant GIST achieve clinical benefit with imatinib mesilate, an orally administered selective inhibitor of the tyrosine kinase receptor. We treated a 43-year-old male patient suffering from a gastric GIST diagnosed during a surgical emergency operation for peritonitis caused by gastric perforation. At the time of the first operation the patient had lost 10 kg body weight over the previous months and was seriously cachectic. During the emergency operation the perforation was sutured. The biopsy results showed the presence of CD1 17 (c-kit) and CD34 markers. A total body CT scan documented the substantial size of the gastric wall lesion, an increased volume of abdominal lymph nodes and compression of the splenic vein with alternative collateral circulation. The liver presented no less than 5 large metastases distributed in both the left and right lobes. There was also a pulmonary metastasis. Because of frequent spontaneous bleeding and starvation the patient was seriously anaemic. Considering the action mechanism of imatinib and the extent of the lesion we decided to perform a total gastrectomy procedure. At the time of the operation the stomach seemed to have a modified volume and shape: it appeared to be divided into two sacs, the larger and deeper of which was the original gastric cavity, while the superficial, smaller one seemed to be a protrusion of the organ. The stomach was indistinguishable from the spleen, the transverse colon and the distal pancreatic tract. The neoplasm was directly linked to the left liver and to the inferior diaphragmatic surface. We performed total gastrectomy and resection of the tail of the pancreas, the spleen, and the transverse colon all in one and the same session. The patient was discharged on postoperative day 8 and commenced imatinib therapy 30 days after the operation with 4 tablets per day. In the following months the patient repeated the CT scan to monitor the progressive volume reduction of the liver and lung lesions and a PET scan confirmed that the lesions were not active; the patient experienced a 13 kg body weight increase. One year after the operation the outcome appears to be lasting and the patient has tolerated the drug treatment well. PMID- 15832749 TI - Pneumatosis cystoides of the right colon: a possible source of misdiagnosis. Report of a case. AB - Pneumatosis cystoides intestinalis is a rare condition that can be located in any part of the gastrointestinal tract. It is usually associated with a wide variety of gastrointestinal or pulmonary diseases. The primitive form is much less frequent and usually involves the left colon. The pathogenesis of pneumatosis cystoides intestinalis is still unclear. The mechanical theory, which is the most accepted explanation, postulates that gas is forced into the bowel wall by breaks in the mucosa; this is more likely to occur when the intraluminal pressure is higher, as happens in obstructive conditions, during endoscopies, or during infections from gas-forming bacteria. Pneumatosis cystoides is often asymptomatic, representing an occasional finding during investigations for other abdominal conditions. Complications occur in about 3% of cases and include obstruction, intussusception, volvulus, haemorrhage and intestinal perforation. When presenting acutely or in association with other abdominal conditions the differential diagnosis is rarely a problem. More important is to diagnose asymptomatic primitive submucosal pneumatosis of the colon, in order to avoid unnecessary intestinal resections. The Authors present the case of a patient with pneumatosis coli who underwent laparotomy for a suspected colonic lipomatosis of the right colon. PMID- 15832751 TI - [Therapy of large varices. There is more than elastics and knives]. PMID- 15832752 TI - [Chronic inflammatory bowel diseases. Recurrence prevention = carcinoma prevention?]. PMID- 15832753 TI - [Survey of colleagues. How do you motivate your helpers?]. PMID- 15832754 TI - [The father has main branch stenosis. Are the sons to be sent for heart catheterization at the same time?]. PMID- 15832755 TI - [Lipophages of Peter Hans Hofschneider. Slender by viruses]. PMID- 15832756 TI - [A physician's essential competence which can be learned. Are you a good listener?]. PMID- 15832757 TI - [Instead of endless discussions on what constitutes optimal treatment of hypertension--identify and treat at-risk patients]. PMID- 15832758 TI - [How to detect impending cardiac insufficiency]. AB - The characteristic leading symptoms of heart failure are a reduction in physical performance, dyspnea and the development of edema. At this point, such examinations as an ECG and determination of various laboratory parameters become mandatory. For the differential diagnostic exclusion of a pulmonary cause of respiratory distress, lung function tests and chest X-rays are needed. If heart failure is suspected clinically, echocardiography is the most important noninvasive method for identifying a cardiac functional disorder. With the aid of this examination, systolic function, diastolic filling, myocardial thickness and valve function can be evaluated. Cardiac catheterization is emportant to determine the underlying causes of heart failure and to plan therapy. PMID- 15832759 TI - [Stress and myocardial infarction]. AB - Most people throughout the world die from the consequences of cardiovascular disease. Stress and psychosocial burdens have, in the past, been underestimated with regard to the importance of their impact on the development and course of such diseases. In the INTERHEART study, psychosocial burdens occupy third place among the risk factors for developing acute myocardial infarction. The relevance of these factors is underscored by more recent studies, also with regard to the prognosis in already manifest CAH. The causes of mental stresses may be intrapsychic problems (e.g. depression). The roots may, however, also be found in the private sphere or at the workplace. On the basis of specific history-taking, relevant risk constellations can be identified for a comparatively low expenditure of time. Specific therapeutic approaches aimed at reducing and coping with stress may, in future, help prevent diseases of the heart and lower the risk of contracting a myocardial infarction. PMID- 15832761 TI - [Sexual problems in women after surgical interventions]. AB - Sexual disorders following operative interventions have a variety of causes. Apart from functional disturbances (e.g. as a result of the formation of scar tissue, hormone deficiencies, etc.), problems arising from the psychological problem of coping with the intervention itself also play an important role. A major course of conflict lies in communication problems between the partners involved. Potential sexual disorders should be discussed in the preoperative period, with the aim of enabling the patient to come to terms with the postoperative situation. However, it remains a task of the physician after the operation to actively support the patient in dealing with the traumatization she has experienced. PMID- 15832760 TI - [Old or new antihypertensives--which are better?]. AB - Some of the reported findings of numerous studies on the treatment of hypertension are still giving rise to heated discussions. The result is conflicting recommendations and uncertainty among care-providing physicians. Today, the substances from the group of more recent hypertensive agents (calcium antagonists, ACE-inhibitors and angiotensin 1 receptor blockers) together with the classical agents (diuretics and beta blockers) are recognized as equally justifiable as the five agents of first choice in the treatment of uncomplicated hypertension. If, however, accompanying diseases are present, the choice of primary medication depends on the respective risk (diabetes, etc.) of the individual patient. In many cases, combination treatment should be considered from the very beginning. PMID- 15832762 TI - [Conclusions from CHARM interview with Prof. Veselin Mitrovich, Bad Nauheim. Strong evidence: more help for weak hearts]. PMID- 15832763 TI - [Fatal decision. Court encourages practice fee refusal]. PMID- 15832764 TI - [How much physician's work time is still really worth? Arbitration office firmly puts the point value at 1.5 cents]. PMID- 15832765 TI - [Look diagnosis. Grown over the years. Unilateral hamster cheek]. PMID- 15832766 TI - European Directive 2004/23/EC of 31 March 2004 on tissue banks, and Italian law. PMID- 15832767 TI - Evaluation of the prevalence of human papillomavirus and Epstein-Barr virus in esophageal squamous cell carcinomas. AB - The aim of this study was to determine the possible involvement of human papillomavirus and Epstein-Barr virus in esophageal squamous cell cancer (ESCC) carcinogenesis in the Greek population. DNA was extracted from 30 ESCC and 27 normal esophageal specimens and screened for HPV type-specific or EBV infection by PCR-based assay. Seventeen out of 30 ESCC specimens (56%) were found positive for HPV DNA, of which 15 (88%) were typed as HPV-18 infected, one (5.9%) as HPV 16 infected, and one (5.9%) as infected by an HPV type different from the studied HPV-6, 11, 16, 18 and 33 subtypes. Six of the 27 normal esophageal specimens (22.2%) were positive for HPV infection, five typed as HPV-18 (83.3%) and one as HPV-16 (16.7%). All samples were negative for EBV genome detection as assessed by the PCR assay. No statistically significant correlation was found between the HPV status of the tumor samples and clinical parameters including sex, age of the patients, tobacco or alcohol use, differentiation grade of the lesions and stage of the disease. In conclusion, our findings indicate a statistically significant (p<0.001) overall association between ESCC and HPV infection, mostly related to the HPV-18 subtype, in the Greek population. PMID- 15832768 TI - Tyrosinase mRNA RT-PCR analysis as an additional diagnostic tool for the identification of melanoma cells in biological fluid samples other than blood: a preliminary report. AB - Reverse transcription polymerase chain reaction (RT-PCR) of tyrosinase mRNA has been applied for the detection of melanoma cells in the peripheral blood, lymph nodes and bone marrow of melanoma patients. We evaluated the diagnostic accuracy of RT-PCR in comparison to standard cytology and immunocytochemistry (ICC) for the identification of melanoma cells in biological fluids other than blood. Tyrosinase expression was evaluated together with standard cytology and ICC (anti S100, HMB-45 and Melan-A antibodies) in biological fluid samples collected from 17 melanoma patients according to the site of metastatic involvement or clinical suspicion (eight cerebrospinal fluid (CSF) samples; three pleural effusions; four ascites; one bile sample, one pericardial effusion); 17 samples collected from patients with non-melanoma metastatic cancer were used as controls. Tyrosinase expression in the biological fluid sample was compared with the expression determined at the same time in peripheral blood. Positive tyrosinase expression was found in 12/17 melanoma and 3/17 non-melanoma cancer patients. Cytology/ICC showed the presence of neoplastic cells in only 7/12 melanoma samples with positive tyrosinase expression: radiological evidence of disease involvement was found in all these patients (three meningeal, two pleural, two peritoneal). Clear cut radiological evidence of disease involvement at the sampling site was found in the five patients with negative cytology/ICC and positive RT-PCR (one CSF; four serous membrane effusions); all patients died of disease progression within four months of sampling. The five patients who were negative for both cytology/ICC and RT-PCR did not show any clinical evidence of disease recurrence at the sampling site. Only five of the 12 metastatic patients with positive tyrosinase expression in biological fluid showed positivity for tyrosinase in the peripheral blood. These preliminary results suggest that the analysis of biological fluids other than blood could be considered as a new potential clinical field of application for the tyrosinase mRNA assay. PMID- 15832769 TI - Transcriptional inactivation of p53, Bax, Bcl-2 and Mdm2 correlates with malignant transformation of the uterine cervix. AB - Deregulation of the apoptotic machinery plays a major role in cell death, cellular transformation and cancer. p53, Bcl-2, Bcl-XL, Bax and Mdm2 mRNA expression patterns were evaluated in tissue samples with cervical intraepithelial neoplasia (CIN) and cervical cancer compared to those of normal cervical tissues, and correlated with the underlying cervical lesions. Transcript levels of the above genes were assessed by RT-PCR analysis in a total of 44 cervical specimens. p53, Bcl-2, Bax and Mdm2 transcript levels were significantly different in the normal, CIN and cancer specimen groups (p=0.003, p=0.009, p=0.040 and p=0.001, respectively). Specifically, p53, Bax and Bcl-2 exhibited substantially lower transcript levels in CIN lesions compared to controls, whereas Bax mRNA levels showed a significant decrease in cancer compared to normal specimens. Mdm2 mRNA expression was considerably lower in cancer than in CIN lesions or normal cervix. High-grade squamous intraepithelial lesions exhibited lower p53 and Bcl-2 mRNA levels than controls (p=0.002, p=0.016). Coexpression analysis revealed more correlations between the above apoptosis related molecules in normal tissues compared to CIN or cancer specimens. p53 showed significant coexpression with Bax, Bcl-2 and Mdm2 (p=0.040, p=0.013 and p=0.015, respectively) in normal cervical specimens. Bax and Bcl-XL mRNA expression was negatively correlated. Mdm2 transcriptional levels correlated significantly with those of Bax, Bcl-XL and Bcl-2. Our findings show that p53, Bax, Bcl-2 and Mdm2 mRNA expression levels correlate with the malignant transformation of the uterine cervix. mRNA coexpression patterns of the members of the pro- and anti-apoptotic family examined in cervical carcinogenesis were found to be disrupted in CIN and cancer, as already demonstrated at the protein level. PMID- 15832770 TI - A new nested primer pair improves the specificity of CK-19 mRNA detection by RT PCR in occult breast cancer cells. AB - Reverse transcription polymerase chain reaction (RT-PCR) of cytokeratin-19 (CK 19) has been widely used to detect small numbers of circulating malignant epithelial cells in the bone marrow or the peripheral blood of patients with breast cancer. However, a high percentage of false positive results has been recorded and conflicting reports question the clinical relevance of this technical approach. We demonstrate that the use of a new nested primer pair for CK-19 RT-PCR avoids false positive results without affecting the sensitivity of the assay. Our experiments were carried out using MCF-7 cells alone or mixed with peripheral blood mononuclear cells (PBMNC) of healthy donors. The results were also validated in a large series of healthy donors and in a preliminary study on a limited number of patients with breast cancer, thus suggesting that our assay is feasible for application in the clinical evaluation of occult malignant epithelial cells. PMID- 15832771 TI - Comparison of four different assays for determination of serum S-100B. AB - BACKGROUND: S-100B determination has been shown to be clinically useful in the management of melanoma patients. After the development of a test for determination of the isoforms S100-A1B and S100-BB in serum (S-100B), several sensitive assays for the detection of serum S-100B have become available. We compared four S-100B assays, two automated (LIAISON Sangtec 100 and Elecsys S100) and two manual ones (Sangtec 100 ELISA and CanAg S100 EIA), with respect to clinical data, reference values and correlation. METHODS: In a total of 280 samples from 155 melanoma patients and 98 healthy individuals S-100B values were measured simultaneously with the different assays. RESULTS: The inter and intra coefficients of variation were best for the automated assays. The functional sensitivity of both manual assays was 0.15 microg/L. Method comparison revealed satisfactory correlation coefficients of 0.9 or higher, but the slopes ranged from 0.29 to 3.36. Except for the Sangtec 100 ELISA, the linearity between the assays was acceptable. The overall sensitivity for melanoma ranged from 37% (Elecsys S100) to 47% (LIAISON Sangtec 100) and the sensitivity increased with stage. ROC curves showed the best accuracy for the LIAISON Sangtec 100 assay. CONCLUSIONS: All assays gave satisfactory results, but it is advisable to improve the performance of the manual assays for better sensitivity. Agreement about an international reference standard is needed. PMID- 15832772 TI - Diagnostic utility of CYFRA 21-1 and CEA assays in pericardial fluid for the recognition of neoplastic pericarditis. AB - A positive cytology result in pericardial fluid is the gold standard for recognition of malignant pericardial effusion. Unfortunately, in 30-50% of patients with malignant pericardial effusion cytological examination of the pericardial fluid is negative. Tumor marker assessment in pericardial fluid may help to recognize malignant pericardial effusion. The aim of our study was to estimate the value of CYFRA 21-1 and CEA measurement in pericardial fluid for the recognition of malignant pericardial effusion. To our knowledge this is the first study on CYFRA 21-1 assessment in pericardial effusion. The examined group consisted of 50 patients with malignant pericardial effusion and 34 patients with non-malignant pericardial effusion. Median CEA concentrations in malignant pericardial effusion and non-malignant pericardial effusion were 80 ng/mL (0-317) and 0.5 ng/mL (0-18.4), respectively (p<0.001). Median CYFRA 21-1 concentrations in malignant pericardial effusion and non-malignant pericardial effusion were 260 ng/mL (5.3-10080) and 22.4 ng/mL (1.87-317.6), respectively (p<0.001). The optimal cutoff value for CYFRA 21-1 in pericardial effusion was 100 ng/mL. CYFRA 21-1 >100 ng/mL or CEA >5 ng/mL were found in 14/15 patients with malignant pericardial effusion and negative pericardial fluid cytology. We therefore strongly recommend the use of CYFRA 21-1 and/or CEA in addition to pericardial fluid cytology for the recognition of malignant pericardial effusion. PMID- 15832773 TI - Cathepsins B and L and their inhibitors stefin B and cystatin C as markers for malignant progression of benign meningiomas. AB - Meningiomas are, in general, slowly growing benign tumors attached to the dura mater and composed of neoplastic meningothelial (arachnoidal) cells. They have a wide range of histopathological appearances and are classified, according to the aggressiveness of their growth and the risk of recurrence, as WHO grade I (benign) meningiomas, WHO grade II (atypical) meningiomas and WHO grade III anaplastic (malignant) meningiomas. As invasion of normal tissue may occur in all grades, independent biological markers are needed to identify the more aggressive and recurrent meningiomas. The lysosomal cysteine proteinases, cathepsins B and L, have been associated with tumor invasiveness and the aim of this study was therefore to evaluate them, together with their endogenous inhibitors stefin B and cystatin C, as potential markers for the aggressiveness of meningiomas. The expression of cathepsins B and L and their inhibitors stefin B and cystatin C in 21 benign (grade I) and 9 atypical (grade II) meningiomas has been compared by immunohistochemical staining, QRT-PCR and Northern blot analysis. The protein levels of cathepsins B (p=0.050) and L (p=0.019) were found to be significantly higher in atypical than in benign meningiomas. In contrast, their mRNA levels did not differ, indicating that the synthesis of cathepsins was accelerated at the translational level. Protein and mRNA levels of stefin B (p= 0.007), but not cystatin C, were significantly lower in atypical compared with benign meningiomas. The expression of cathepsins and inhibitors was not different between central and peripheral meningioma tissue or between histological subtypes of meningiomas, with the exception of cathepsin L, the level of which was significantly lower in transitional meningiomas. We conclude that higher protein levels of cathepsins B and L and lower mRNA levels of stefin B are potential diagnostic markers for invasive and aggressive behavior of meningiomas. The diagnostic and prognostic value for relapse of meningioma needs to be confirmed in a larger population of patients. PMID- 15832774 TI - The favorable prognostic significance of surgery-induced hyperprolactinemia in node-positive breast cancer patients: ten-year disease-free survival results. AB - It has been shown that each manipulation of the mammary region, including breast surgery, may stimulate prolactin secretion. However, it has also been observed that in more than 50% of breast cancer patients surgical removal of the tumor is not followed by enhanced prolactin secretion. This might be indicative of an altered psychoneuroendocrine control of the mammary gland, which could lead to the onset of more biologically aggressive breast cancer. In fact, surgery-induced hyperprolactinemia has been proven to be associated with a better prognosis in terms of survival in node-negative breast cancer patients. The present study was performed to investigate the impact of postoperative hyperprolactinemia on the disease-free survival (DFS) of breast cancer patients with axillary node involvement. The study included 100 consecutive node-positive breast cancer patients who were followed for at least 10 years. Surgery-induced hyperprolactinemia occurred in 45/100 (45%) patients without any significant correlation with the main prognostic variables including number of involved nodes and ER status. The two groups of patients received the same adjuvant therapies. After a median follow-up of 151 months, the recurrence rate in patients with surgery-induced hyperprolactinemia was significantly lower than in patients with no postoperative hyperprolactinemia (23/45 vs 43/55, p<0.01). Moreover, DFS was significantly longer in hyperprolactinemic patients than in patients who had no enhanced secretion of prolactin postoperatively. In agreement with the results described previously in node-negative breast cancer, our study demonstrates the favorable prognostic significance of surgery-induced hyperprolactinemia in terms of DFS duration also in breast cancer patients with axillary node involvement, independent of the other well-known prognostic variables, thereby confirming that the psychoneuroendocrine status of cancer patients may influence the prognosis of their disease. PMID- 15832775 TI - Altered expression of the DNA mismatch repair proteins hMLH1 and hMSH2 in cutaneous dysplastic nevi and malignant melanoma. AB - Molecular alterations in the mismatch repair system suggest that this mechanism may be important in the evolution of cutaneous melanoma. Our current study evaluated the expression of two mismatch repair proteins, hMLH1 and hMSH2, in dysplastic nevi (DN) and cutaneous melanoma (CM). Immunohistochemical staining of these proteins was performed on 55 CM and 30 DN specimens. The staining results were divided into three groups: negative, partially positive and strongly positive. Normal adjacent skin cells served as an internal control for positive immunostaining. Altered immunoreactivity of one of the proteins was found in four (13.4%) DN and seven (12.7%) CM. Lack of staining for hMLH1 was observed in two (6.7%) cases of DN and five (9.1%) cases of CM; staining for hMSH2 was absent in two (6.7%) of the DN and two (3.6%) of the CM specimens. Partially positive staining was found in 33.3% and 53.3% for hMLH1 and hMSH2, respectively, in DN, and in 54.5% and 69.1%, respectively, in CMM. Our study shows that complete or partial loss of MMR protein expression occurs in a subset of both DN and CM and may represent a distinct pathway in the development of some DN and CM. PMID- 15832776 TI - Glutathione S-transferase M1, T1 and P1 polymorphisms and bladder cancer risk in Egyptians. AB - Previous studies suggest that bladder cancer risk may vary with GST genotype but these results are inconsistent. The aim of this study was to explore whether GSTM1, GSTT1 and GSTP polymorphisms were associated with increased bladder cancer risk in an Egyptian population. GSTM1, GSTT1 and GSTP1 genotype frequencies were determined in bladder cancer cases (n=72) and healthy controls with no history of malignancies (n=82) using PCR-based techniques. The GSTT1*2 genotype was particularly associated with increased risk (OR 2.71, 95%CI 1.27-5.73) and the GSTM1*2 genotype to a lesser extent (OR 1.63, 95%CI 0.85-3.10). 18.1% of cases but only 7.3% of controls were GSTP1*B*B homozygotes (OR 2.38, 95%CI 0.83-6.87). The presence of two or more a priori at-risk genotypes was associated with increased bladder cancer risk (OR 2.42; 95%CI 1.47-3.97). These results suggest that polymorphisms in the GST genes are associated with increased risk of bladder cancer among Egyptians. PMID- 15832777 TI - Agreement between observers: its measure on a quantitative scale. PMID- 15832778 TI - [Coronary disease patient has already LDL of 100 mg/dl. Why lipid lowering is still worthwhile]. PMID- 15832779 TI - [New goals in secondary prevention. "It does not make sense to stop at 100 mg/dl LDL"]. PMID- 15832780 TI - [Hypertension therapies compared. Are the new nevertheless better than the old?]. PMID- 15832781 TI - [Hypertension therapy with beta blocker plus diuretic. "We are at the point of leaving behind an era"]. PMID- 15832782 TI - [Female, young, healthy. ASS does not protect from myocardial infarct]. PMID- 15832783 TI - [After the disappointing ASS primary prevention trial. "Holding back in women"]. PMID- 15832784 TI - [In infantile diarrhea fast rehydration and realimentation. The tea stop is out!]. PMID- 15832785 TI - [Won't the women be accepted seriously? "Menopause discomfort--what cynicism"]. PMID- 15832786 TI - [Section without medical indications. The physician has to refuse it]. PMID- 15832787 TI - [Please not with dirty fingernails!]. PMID- 15832788 TI - [Against severest pain. A new nerve stimulator tested]. PMID- 15832789 TI - [Screaming children -- refusing to eat -- night noise -- truants. Parents on the brink of nervous breakdown]. PMID- 15832790 TI - [Regulatory disorders in early childhood. Family physician counseling for crying, sleeping and feeding disorders]. AB - Early childhood regulatory disorders account for the most common behavioral abnormalities of the infant and young child that result from regulatory problems associated with age-specific adaptational and developmental tasks. Increasing empirical evidence indicates that these problems need to receive full attention. If left to themselves, such disorders impair the early parent-child relationship, and increase the risk for disturbances later on in life. In most cases, the family doctor is the first person to be consulted by the infant's parents who are unable to cope with excessive crying, sleeping or feeding disorders. A range of proven methods are available to support counseling on child development in the physician's office. PMID- 15832791 TI - [The importance of early traumatic experiences for the development of the infant's brain]. AB - Learning and social and emotional experiences have an impact on gene activity, and modify the growth and function of nerve cells. As a result, and depending on the interactive experiences within relationships, motoric, cognitive and emotional skills, as well as behavior of the infant are influenced. If the child fails to experience a dependable relationship, the symptoms of mental disturbance often develop (e.g. ADHD, among others). Further factors that impact on brain development and lead to posttraumatic stress disorder (PTSD) are deprivation, physical violence and sexual abuse in early childhood. PMID- 15832792 TI - [Mental disorders in puberty]. AB - Puberty is characterized by a complexity of different developmental tasks. During this period of sometimes dramatic changes, the adolescent must find a new balance between personal experiences and the demands made by the environment. In this period of development, some 18% of all adolescents experience a mental crisis, and 5% need specialist treatment. Such mental disorders in puberty as anorexia nervosa, depression or suicidality can be classified on the basis of their development and course. They must be taken seriously and treated as early as possible. PMID- 15832793 TI - [Pitfalls in the diagnosis and treatment optimization of type 2 diabetes]. AB - The diagnosis of diabetes mellitus would appear a simple matter. However, potential pitfalls in clinical practice need to be avoided, and this requires knowledge and attention. An initially pathological oral glucose tolerance test should be repeated before establishing a final diagnosis, since such necessary preconditions as a 10 to 16 hours fast, or alcohol abstinence, are difficult to monitor in the clinical setting. Accurate glucose testing requires appropriate sample preparation and handling. Further pitfalls may be encountered during treatment: HbA1c assessment is associated with certain limitations and does not permit the estimation of glucose variations. To establish a differential diagnoses between type 1 (LADA) and type 2 diabetes in older patients GAD must be measured. New biochemical markers such as adiponectin and intact proinsulin may facilitate treatment decisions and monitoring in patients with type 2 diabetes. PMID- 15832794 TI - [Endocranial complications in otitis media: what you need to remember]. AB - Thanks to the potency of antibiotics, life-threatening complications of otitis media are rare. In many cases, however, the symptomatology is veiled, and the course of the disease tends to be protracted. Secondary symptoms may preponderate and often mask those associated with the ear itself. If a case of otitis media fails to clear up, or mastoiditis develops, or sensorineural deafness, vertigo, headaches, seizures, or meningism occur, the possibility of an otogenous complication must immediately be considered and the patient submitted to diagnostic imaging without delay, so as to ensure early surgical treatment of a possible lesion in the ear. PMID- 15832795 TI - [Many colleagues would not themselves gladly clean up. Is your waiting room a catastrophe, too?]. PMID- 15832796 TI - [New EBM. ECG in emergency services is accountable, too]. PMID- 15832797 TI - [Gait disorders in the aged. What to do when your patient's gait is impaired?]. PMID- 15832798 TI - [Patient care in Cameroon. As medical fee two dead monkeys]. PMID- 15832799 TI - [Look diagnosis. arched erythema]. PMID- 15832800 TI - The European Constitution, establishing a core of human rights as applied to the fields of medicine and biology. PMID- 15832801 TI - Body transformations. PMID- 15832802 TI - Research and use of stem cells. AB - Stem cells have enormous potential in medicine. Stem cell research is allowedsometimes just for to a limited extent in a large minority of countries. Nevertheless, the research and use of stem cells should be encouraged. PMID- 15832803 TI - Contemporary issues in the regulation of artificial reproduction and embryology in the UK. AB - The Human Fertilisation and Embryology Act 1990 was one of the most comprehensive and flexible vehicles for regulation of artificial reproduction and embryology of its area. However, its serviceability has recently been brought into question by both the materialisation of unforeseen dilemmas and the response of legal actors to these dilemma's which has often been ethically and democratically deficient. Nowhere is this better demonstrated than in the reproach of the regulatory authority and judiciary to the use pre-implantation genetic diagnosis and cloning by cell nuclear replacement. PMID- 15832804 TI - Privacy and intra-familiy communication of genetic information. AB - The new knowledge (and predictions) created by DNA tests and the family nature of genetic information has already lead to a new problem: the intra-familiar communication of genetic data. This raises questions such as the following. Is there a duty to inform in cases when treatment is possible and the patient does not permit disclosure of genetic results to relatives? Is there an obligation to warn or merely an authorization (that could be used or not)? Could privacy protection be maintain as an individual interest but with some justified violations? A balance needs to be establishes between the interest of privacy and the need to disclose secret information. PMID- 15832805 TI - Icelandic. Decision of the Supreme Court on the protection of privacy with regard to the processing of Health Sector Databases. Attorney at Law vs The State of Iceland. AB - Mr. R appealed for a decision by the Court to overturn the refusal of the Medical Director of Health to her request that health information in medical records pertaining to herdeceased father should not be entered into the Health Sector Database. Furthermore, she called for recognition of her right to prohibit the transfer of such information into a database. Article 8 of Act No 139/1998 on a Health Sector Database provides for the right of patients to refuse permission, by notification to the Medical Director of Health, for information concerning them to be entered into the Health Sector Database. The Court concluded that R could not exercise this right acting as a substitute of her deceased father, but it was recognised that she might, on the basis of her right to protection of privacy, have an interest in preventing the transfer of health data concerning her father into the database, as information could be inferred from such data relating to the hereditary characteristics of her father which might also apply to herself. It was revealed in the course of proceedings that extensive information concerning people's health is entered into medical records, e.g. medical treatment, life-style and social conditions, employment and family circumstances, together with a detailed identification of the person that the information concerns. It was recognised as unequivocal that the provisions of Paragraph 1 of Article 71 of the Constitution applied to such information and guaranteed to every person the right to protection of privacy in this respect. The Court concluded that the opinion of the District Court, which, inter alia, was based on the opinion of an assessor, to the effect that so-called one-way encryption could be carried out in such a secure manner that it would be virtually impossible to read the encrypted data, had not been refuted. It was noted, however, that Act No. 139/1998 provides no details as to what information from medical records is required to be encrypted in this manner prior to transfer into the database or whether certain information contained in the medical records will not be transferred into the database. The documents of the case indicate that only the identity number of the patient would be encrypted in the database, and that names, both those of the patient and his relatives, as well as the precise address, would be omitted. It is obvious that information on these items is not the only information appearing in the medical records which could, in certain cases, unequivocally identify the person concerned. Act No. 139/1998 also provides for authorisation to the licensee to process information from the medical records transferred into the database. The Act stipulates that certain specified public entities must approve procedures and process methods and monitor all queries and processing of information in the database. However, there is no clear definition of what type of queries will be directed to the database or in what form the replies to such queries will appear. The Court concluded that even though individual provisions of Act No 139/1998 repeatedly stipulate that health information in the Health Sector Database should be non-personally identifiable, it is far from adequately ensured under statutory law that this stated objective will be achieved. In light of the obligations imposed on the legislature by Paragraph 1 of Article 71 of the Constitution, the Court concluded that various forms of monitoring of the creation and, operation of the database are no substitute in this respect without foundation in definite statutory norms. In light of these circumstances, and taking into account the principles of Icelandic law concerning the confidentiality and protection of privacy, the Court concluded that the right of R in this matter must be recognised, and her court claims, therefore, upheld. PMID- 15832806 TI - [Social perceptions on genomics in four Latin American countries. Ethical-legal implications]. AB - The authors analyze under an ethical and legal perspective the consequences and anxieties generated by the human genome project in the population of four Latin American countries: Argentine, Chile, Mexico and Peru, through bibliographical analysis and interviews done to biomedical researches, lawyers and legislators, students and lay civilians. PMID- 15832807 TI - The animal farm philosophy of genetic discrimination. AB - The paper by Dr. Gregor Wolbring addresses the issue of genetic discrimination from disabled people's rights perspective asking a) what the interpretation of genetic discrimination and the scope of Anti Genetic discrimination laws and law proposals is and b) whether the scope and interpretation of genetic discrimination and Anti Genetic discrimination laws and law proposal lead to more protection for-or increased discrimination against- disabled people" PMID- 15832808 TI - [Embryo selection and bioethical principles]. AB - This paper analyzes, from the point of view of the principles of Bioethics, the problems of embryonic selection with the purpose of serving as donors of future transplant for their brothers seriously ill. Analyzed the principles of Bioethics of no malice, charity, autonomy and justice in connection with the matter explained, the author defends its moral fairness and he explains the necessity of proceeding to a legal reform which permits to accommodate in the Spanish Legal Code to this kind of embryonic selection, since nowadays laws into effect prohibit this practice with the purpose discussed here. PMID- 15832809 TI - Additional protocol to the convention for the protection of human rights and dignity of the human being with regard to the application of biology and medicine, on biomedical research. PMID- 15832810 TI - Induction of phase I, II and III drug metabolism/transport by xenobiotics. AB - Drug metabolizing enzymes (DMEs) play central roles in the metabolism, elimination and detoxification of xenobiotics and drugs introduced into the human body. Most of the tissues and organs in our body are well equipped with diverse and various DMEs including phase I, phase II metabolizing enzymes and phase III transporters, which are present in abundance either at the basal unstimulated level, and/or are inducible at elevated level after exposure to xenobiotics. Recently, many important advances have been made in the mechanisms that regulate the expression of these drug metabolism genes. Various nuclear receptors including the aryl hydrocarbon receptor (AhR), orphan nuclear receptors, and nuclear factor-erythoroid 2 p45-related factor 2 (Nrf2) have been shown to be the key mediators of drug-induced changes in phase I, phase II metabolizing enzymes as well as phase III transporters involved in efflux mechanisms. For instance, the expression of CYP1 genes can be induced by AhR, which dimerizes with the AhR nuclear translocator (Arnt), in response to many polycyclic aromatic hydrocarbon (PAHs). Similarly, the steroid family of orphan nuclear receptors, the constitutive androstane receptor (CAR) and pregnane X receptor (PXR), both heterodimerize with the retinoid X receptor (RXR), are shown to transcriptionally activate the promoters of CYP2B and CYP3A gene expression by xenobiotics such as phenobarbital-like compounds (CAR) and dexamethasone and rifampin-type of agents (PXR). The peroxisome proliferator activated receptor (PPAR), which is one of the first characterized members of the nuclear hormone receptor, also dimerizes with RXR and has been shown to be activated by lipid lowering agent fibrate-type of compounds leading to transcriptional activation of the promoters on CYP4A gene. CYP7A was recognized as the first target gene of the liver X receptor (LXR), in which the elimination of cholesterol depends on CYP7A. Farnesoid X receptor (FXR) was identified as a bile acid receptor, and its activation results in the inhibition of hepatic acid biosynthesis and increased transport of bile acids from intestinal lumen to the liver, and CYP7A is one of its target genes. The transcriptional activation by these receptors upon binding to the promoters located at the 5-flanking region of these CYP genes generally leads to the induction of their mRNA gene expression. The physiological and the pharmacological implications of common partner of RXR for CAR, PXR, PPAR, LXR and FXR receptors largely remain unknown and are under intense investigations. For the phase II DMEs, phase II gene inducers such as the phenolic compounds butylated hydroxyanisol (BHA), tert-butylhydroquinone (tBHQ), green tea polyphenol (GTP), (-)-epigallocatechin-3-gallate (EGCG) and the isothiocyanates (PEITC, sulforaphane) generally appear to be electrophiles. They generally possess electrophilic-mediated stress response, resulting in the activation of bZIP transcription factors Nrf2 which dimerizes with Mafs and binds to the antioxidant/electrophile response element (ARE/EpRE) promoter, which is located in many phase II DMEs as well as many cellular defensive enzymes such as heme oxygenase-1 (HO-1), with the subsequent induction of the expression of these genes. Phase III transporters, for example, P-glycoprotein (P-gp), multidrug resistance-associated proteins (MRPs), and organic anion transporting polypeptide 2 (OATP2) are expressed in many tissues such as the liver, intestine, kidney, and brain, and play crucial roles in drug absorption, distribution, and excretion. The orphan nuclear receptors PXR and CAR have been shown to be involved in the regulation of these transporters. Along with phase I and phase II enzyme induction, pretreatment with several kinds of inducers has been shown to alter the expression of phase III transporters, and alter the excretion of xenobiotics, which implies that phase III transporters may also be similarly regulated in a coordinated fashion, and provides an important mean to protect the body from xenobiotics insults. It appears that in general, exposure to phase I, phase II and phase III gene inducers may trigger cellular "stress" response leading to the increase in their gene expression, which ultimately enhance the elimination and clearance of these xenobiotics and/or other "cellular stresses" including harmful reactive intermediates such as reactive oxygen species (ROS), so that the body will remove the "stress" expeditiously. Consequently, this homeostatic response of the body plays a central role in the protection of the body against "environmental" insults such as those elicited by exposure to xenobiotics. PMID- 15832811 TI - Open channel block of hKv1.5 by psoralen from Heracleum moellendorffii Hance. AB - A furocoumarin derivative, psoralen (7H-furo[3,2-g][1]benzopyran-7-one), was isolated from the n-hexane fraction of Heracleum moellendorffii Hance. We examined the effects of psoralen on a human Kv1.5 potassium channel (hKv1.5) cloned from human heart and stably expressed in Ltk- cells. We found that psoralen inhibited the hKv1.5 current in a concentration-, use- and voltage dependent manner with an IC50 value of 180 +/- 21 nM at +60 mV. Psoralen accelerated the inactivation kinetics of the hKv1.5 channel, and it slowed the deactivation kinetics of the hKv1.5 current resulting in a tail crossover phenomenon. These results indicate that psoralen acts on the hKv1.5 channel as an open channel blocker. Furthermore, psoralen prolonged the action potential duration of rat atrial muscles in a dose-dependent manner. Taken together, the present results strongly suggest that psoralen may be an ideal antiarrhythmic drug for atrial fibrillation. PMID- 15832812 TI - Inhibition of interleukin-12 production in mouse macrophages via decreased nuclear factor-kappaB DNA binding activity by myricetin, a naturally occurring flavonoid. AB - Pharmacological inhibition of interleukin-12 (IL-12) production may be a therapeutic strategy for preventing the development and progression of disease in experimental models of autoimmunity. In this study, the effects of myricetin, a naturally occurring flavonoid present in fruits, vegetables and medicinal herbs, on the production of IL-12 were investigated in mouse macrophages stimulated with lipopolysaccharide (LPS). Myricetin significantly inhibited the LPS-induced IL-12 production from both primary macrophages and the RAW264.7 monocytic cell-line in a dose-dependent manner. The effect of myricetin on IL-12 gene promoter activation was analyzed by transfecting RAW264.7 cells with IL-12 gene promoter/luciferase constructs. The repressive effect was 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 myricetin, indicating that myricetin inhibited IL-12 production in LPS-activated macrophages via the down-regulation of NF-kappaB binding activity. PMID- 15832813 TI - Sesquiterpenes from Syneilesis palmata and their cytotoxicity against human cancer cell lines in vitro. AB - The chromatographic separation of the MeOH extract from the aerial parts of Syneilesis palmata led to the isolation of a new sesquiterpene glycoside 4, together with four known compounds. Their structures were characterized to be 4beta,5beta-epoxy-caryophill-8,(15)-ene (1), 3beta-hydroxy-gultin-5-ene (2), 4alpha,5beta-dihydroxy-caryophill-8,(15)-ene (3), (-)-oplopan-4-one-10-alpha-O beta-D-glucose (4) and 3-hexenyl-1-O-beta-D-glucopyranose (5), based on spectroscopic and chemical methods. Compound 2 showed moderate cytotoxicity against five human tumor cell lines in vitro with its ED50 values ranging from 5.90 to approximately 10.83 microg/mL. PMID- 15832814 TI - New cytotoxic sulfated saponins from the starfish Certonardoa semiregularis. AB - Two new sulfated saponins designated as certonardosides P2 and I3 (1 and 2) were isolated from the brine shrimp active fraction of the MeOH extract of the starfish Certonardoa semiregularis. The structures were determined on the basis of spectral analysis. Compounds 1 and 2 were tested for cytotoxicity against five human tumor cell lines (A549, SK-OV-3, SK-MEL-2, XF498, and HCT15), and compound 1 displayed significant cytotoxicity against the SK-MEL-2 skin cancer cell. PMID- 15832815 TI - Cytotoxic constituents of the octocoral Dendronephthya gigantea. AB - A known monoalkyl glycerol ether, (+/-)-1-nonadecyloxy-2,3-propanediol (1) was isolated from the ethyl acetate extract of a soft coral Dendronephthya gigantea as a weakly cytotoxic constituent against four human cancer cell lines, A549, HT 29, HT-1080, and SNU-638. In addition, a known ceramide, (2S,3R,4E,8E)-N hexadecanoyl-2-amino-4,8-octadecadiene-1,3-diol (2), was also isolated as an inactive constituent. This is the first report on the isolation of the compounds 1 and 2 from the octocoral, Dendronephthya species. PMID- 15832816 TI - Potentially bioactive two new natural sesquiterpenoids from the rhizomes of Zingiber zerumbet. AB - Repeated chromatography of the n-hexane-soluble fraction of the MeOH extract of the rhizomes of Zingiber zerumbet led to the isolation of two isomers of 6 methoxy-2E,9E-humuladien-8-one (1 and 2) and stigmast-4-en-3-one. The structures of 1 and 2 were determined by spectroscopic methods including 1 D and 2D-NMR elucidated by analysis of spectroscopic data as well as by comparison with published values. This is the first report on the isolation of compounds 1 and 2 from the nature. Stigmast-4-en-3-one was first isolated from this plant. PMID- 15832817 TI - Flavonoids differentially modulate nitric oxide production pathways in lipopolysaccharide-activated RAW264.7 cells. AB - Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activated RAW264.7 cells. Although flavonol-typed flavonoids (kaempferol and quercetin) more potently scavenged reactivity of nitric oxide (*NO) as well as peroxynitrite (ONOO-) than isoflavones (genistein and genistin), kaempferol, quercetin and genistein showed a little difference in inhibition of both inducible NO synthase expression and NO production, with IC50 values of 13.9, 20.1 and 26.8 microM. However, there was a striking pattern related to structural feature in modulation of LPS-mediated signaling pathways. Thus, flavonols only inhibited transcription factor AP-1 activation, whereas isoflavones suppressed the DNA binding activation of NF-kappaB and C/EBPbeta. Therefore, these data suggest that structural feature may be linked to decide drugs target molecule in LPS-mediated signaling pathways, rather than its potency. PMID- 15832818 TI - Agastache rugosa leaf extract inhibits the iNOS expression in ROS 17/2.8 cells activated with TNF-alpha and IL-1beta. AB - It has been suggested that nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) may act as a mediator of cytokine-induced effects on bone turn over. NO is also recognized as an important factor in bone remodeling, i.e., participating in osteoblast apoptosis in an arthritic joint. The components of Agastache rugosa are known to have many pharmacological activities. In the present study, we investigated the effects of Agastache rugosa leaf extract (ELAR) on NO production and the iNOS expression in ROS 17/2.8 cells activated by a mixture of inflammatory cytokines including TNF-alpha and IL-1beta. A preincubation with ELAR significantly and concentration-dependently reduced the expression of iNOS protein in ROS 17/2.8 cells activated with the cytokine mixture. Consequently, the NO production was also significantly reduced by ELAR with an IC50 of 0.75 mg/mL. The inhibitory mechanism of iNOS induction by ELAR prevented the activation and translocation of NF-kappaB (p65) to the nucleus from the cytosol fraction. Furthermore, ELAR concentration-dependently reduced the cellular toxicity induced by sodium nitroprusside, an NO-donor. These results suggest that ELAR may be beneficial in NO-mediated inflammatory conditions such as osteoporosis. PMID- 15832819 TI - Protection of LLC-PK1 cells against hydrogen peroxide-induced cell death by modulation of ceramide level. AB - Oxidative stress has been reported to elevate ceramide level during cell death. The purpose of the present study was to modulate cell death in relation to cellular glutathione (GSH) level and GST (glutathione S-transferase) expression by regulating the sphingolipid metabolism. LLC-PK1 cells were treated with H2O2 in the absence of serum to induce cell death. Subsequent to exposure to H2O2, LLC PK1 cells were treated with desipramine, sphingomyelinase inhibitor, and N acetylcysteine (NAC), GSH substrate. Based on comparative visual observation with H2O2-treated control cells, it was observed that 0.5 microM of desipramine and 25 mM of NAC exhibited about 90 and 95% of cytoprotection, respectively, against H2O2-induced cell death. Desipramine and NAC lowered the release of LDH activity by 36 and 3%, respectively, when compared to 71% in H2O2-exposed cells. Cellular glutathione level in 500 microM H2O2-treated cells was reduced to 890 pmol as compared to control level of 1198 pmol per mg protein. GST P1-1 expression was decreased in H2O2-treated cells compared to healthy normal cells. In conclusion, it has been inferred that H2O2-induced cell death is closely related to cellular GSH level and GST P1-1 expression in LLC-PK1 cells and occurs via ceramide elevation by sphingomyelinase activation. PMID- 15832820 TI - The difference between nicardipine and its enantiomers on inhibiting vasoconstriction of isolated rabbit thoracic artery. AB - The present study was designed to study the difference effects between nicardipine and its two enantiomers on thoracic artery of rabbit. A high performance liquid chromatographic method was used to prepare the two enantiomers of nicardipine. The thoracic artery of rabbit was removed. The vessels were cut into 3 mm in width and 15 mm in length spiral strips and immersed into tissue baths. The concentration-response curves of nicardipine and its enantiomers were obtained by cumulative administration of the vasoconstrictors. Nicardipine and the enantiomers could shift the dose-response curves of NE, KCl or CaCl2 to right in a nonparallel manner and decrease the maximum effective in a concentration depended manner, respectively. The pD2' value of R-(-)-nicardipine showed significantly effective than that of nicardipine and S-(+)-nicardipine (P<0.01). There was not obviouse difference between the pD2' value of nicardipine and S-(+) nicardipine (P>0.05). The results demonstrate that the stereoselective interaction between R-(-)-nicardipine and L-calcium channel receptor is more stronger than that of S-(+)-nicardipine. PMID- 15832821 TI - Hepatoprotective effect of lactic acid bacteria, inhibitors of beta-glucuronidase production against intestinal microflora. AB - The hepatoprotective activity of lactic acid bacteria (Lactobacillus brevis HY7401, Lactobacillus acidophilus CSG and Bifidobacterium longum HY8001), which inhibited beta-glucuronidase productivity of intestinal microflora, on t-BHP- or CCl4-induced hepatotoxicity of mice were evaluated. These oral administration of lactic acid bacteria lowered beta-glucuronidase production of intestinal microflora as well as Escherichia coli HGU-3. When lactic acid bacteria at a dose of 0.5 or 2 g (wet weight)/kg was orally administered on CCl4-induced liver injury in mice, these bacteria significantly inhibited the increase of plasma alanine transferase and aspartate transferase activities by 17-57% and 57-66% of the CCl4 control group, respectively. These lactic acid bacteria also showed the potent hepatoprotective effect against t-BHP-induced liver injury in mice. The inhibitory effects of these lactic acid bacteria were more potent than that of dimethyl diphenyl bicarboxylate (DDB), which have been used as a commercial hepatoprotective agent. Among these lactic acid bacteria, L. acidophilus CSG exhibited the most potent hepatoprotective effect. Based on these findings, we insist that an inhibitor of beta-glucuronidase production in intestine, such as lactic acid bacteria, may be hepatoprotective. PMID- 15832822 TI - Effect of a new hepatoprotective agent, YH-439, on the hepatobiliary transport of organic cations (OCs): selective inhibition of sinusoidal OCs uptake without influencing glucose uptake and canalicular OCs excretion. AB - The effect of a new hepatoprotective agent, YH-439, on the hepatobiliary transport of a model organic cation (OC), TBuMA (tributylmethylammonium), was investigated. The area under the plasma concentration-time curve (AUC) from time zero to 4 h following iv administration of TBuMA (6.6 micromol/kg) was increased significantly when YH-439 in corn oil (300 mg/kg) was orally administered to rats 24 h prior to the experiment. Nevertheless, the cumulative biliary excretion of TBuMA remained unchanged. As a consequence, the apparent biliary clearance (CLb) of TBuMA was decreased significantly as a result of YH-439 pretreatment, consistent with the fact that the in vivo excretion clearance of TBuMA across the canalicular membrane (CLexc) was not changed by the pretreatment. The in vitro uptake of TBuMA into isolated hepatocytes was decreased by one half by the pretreatment, owing to a decrease in the apparent Vmax and CLlinear, but the Km for the process remained constant. Most interestingly, however, the sinusoidal uptake of glucose, a nutrient, into hepatocytes was not influenced by the pretreatment, suggesting the YH-439 pretreatment specifically impaired the sinusoidal uptake of OCs. Thus, the OC-specific inhibition of hepatic uptake, without influencing the uptake of glucose, a nutrient, appeared to be associated with the hepatoprotective activity of YH-439. PMID- 15832823 TI - Memory enhancing and neuroprotective effects of selected ginsenosides. AB - The effects of ginsenosides Rg3(R), Rg3(S) and Rg5/Rk1 (a mixture of Rg5 and Rk1, 1:1, w/w), which are components isolated from processed Panax ginseng C.A. Meyer (Araliaceae), on memory dysfunction were examined in mice using a passive avoidance test. The ginsenosides Rg3(R), Rg3(S) or Rg5/Rk1, when orally administered for 4 days, significantly ameliorated the memory impairment induced by the single oral administration of ethanol. The memory impairment induced by the intraperitoneal injection of scopolamine was also significantly recovered by ginsenosides Rg3(S) and Rg5/Rk1. Among the three ginsenosides tested in this study, Rg5/Rk1 enhanced the memory function of mice most effectively in both the ethanoland scopolamine-induced amnesia models. Moreover, the latency period of the Rg5/Rk1-treated mice was 1.2 times longer than that of the control (no amnesia) group in both models, implying that Rg5/Rk1 may also exert beneficial effects in the normal brain. We also evaluated the effects of these ginsenosides on the excitotoxic and oxidative stress-induced neuronal cell damage in primary cultured rat cortical cells. The excitotoxicity induced by glutamate or N-methyl D-aspartate (NMDA) was dramatically inhibited by the three ginsenosides. Rg3(S) and Rg5/Rk1 exhibited a more potent inhibition of excitotoxicity than did Rg3(R). In contrast, these ginsenosides were all ineffective against the H2O2- or xanthine/xanthine oxidase-induced oxidative neuronal damage. Taken together, these results indicate that ginsenosides Rg3(S) and Rg5/Rk1 significantly reversed the memory dysfunction induced by ethanol or scopolamine, and their neuroprotective actions against excitotoxicity may be attributed to their memory enhancing effects. PMID- 15832824 TI - Ginsan improved Th1 immune response inhibited by gamma radiation. AB - Gamma radiation causes suppression of the immune function, and immune properties are related to cytokine production. In the present study, the polysaccharide, Ginsan, purified from an ethanol-insoluble fraction of Ginseng (Panax ginseng C.A. Meyer, Araliaceae) water extract was studied to assess its effects on the immunosuppressive activities of gamma radiation. Ginsan was found to stimulate murine normal splenocytes by inducing the mRNA expressions of Th1 and Th2 type cytokines, and also restore the mRNA expression of IFN-gamma, Th1 cytokine, after its inhibition by whole-body gamma irradiation. Therefore, Ginsan was found to restore the T lymphocytes function that had been suppressed by gamma irradiation in allogenic MLR (mixed lymphocyte reactions). However, Ginsan exhibited no excessive stimulatory effects on the control group. The above results indicated that Ginsan may constitute a new noble agent for the improvement of gamma radiation-induced immunosuppression. PMID- 15832825 TI - TERT mRNA expression is up-regulated in MCF-7 cells and a mouse mammary organ culture (MMOC) system by endosulfan treatment. AB - Endosulfan is one of the organochlorine pesticides, which are well-known endocrine disruptors (EDs), and it acts as an estrogen agonist. Estrogen is a group of hormones that play an important role in mammary gland function and are implicated in mammary carcinogenesis. In the present study, we studied the effects of endosulfan on nodule like alveolar lesion (NLAL) formation in mouse mammary gland development using a mouse mammary gland organ culture (MMOC) system. Although endosulfan-treated mammary glands did not form NLALs, more alveolar buds were formed in this group than in the negative control (vehicle treated) group. In addition, telomerase reverse transcriptase (TERT) mRNA expression levels were increased in endosulfan-treated mammary glands in a dose dependent manner. Telomerase can be activated by estrogen, therefore, we examined the effects of endosulfan on telomerase activity, and found that the telomerase activity in estrogen receptor-positive MCF-7 cells was up-regulated by endosulfan treatment. Moreover, this activation was accompanied by the up-regulation of the TERT mRNA expression. Also, transient expression assays using CAT reporter plasmids containing various fragments of the TERT promoter showed that this imperfect palindromic estrogen-responsive element is almost certainly responsible for the transcriptional activation by endosulfan. These results may help elucidate the endocrine disrupting mechanism of endosulfan. PMID- 15832826 TI - Comparison of spectrophotometric, HPLC and chemiluminescence methods for 3 nitrotyrosine and peroxynitrite interaction. AB - We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite induced luminol or lucigenin chemiluminescence were significantly decreased by 3 nitrotyrosine, in concentration-dependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3 nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCl) was also studied via the chemiluminescence assay, where the HOCl-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine. PMID- 15832827 TI - Synthesis and characterization of HPMC derivatives as novel duodenum-specific coating agents. AB - HPMC (Hydroxypropyl methylcellulose) was chemically modified, using maleic anhydrides, to obtain pH-sensitive HPMCAM (Hydroxypropyl methylcellulose acetate maleate) polymers for use as novel duodenum-specific coating agents. The pharmaceutical properties of HPMCAM, such as film forming, acid values, pH sensitive values, water vapor permeability, tensile strength and Tg, were investigated, and found to show good film forming properties. The pH-sensitive values were 3.0 to 3.7. In vitro results demonstrate that HPMCAM could completely suppress drug release within 2h in a simulated gastric fluid (pH 1.2) and rapidly release the drug in a simulated pathological duodenal fluid (pH 3.4). These results indicate that HPMCAM might be a useful material for a duodenum-specific drug delivery system. PMID- 15832828 TI - Development of new reverse micellar microencapsulation technique to load water soluble drug into PLGA microspheres. AB - The objective of this study was to develop a new reverse micelle-based microencapsulation technique to load tetracycline hydrochloride into PLGA microspheres. To do so, a reverse micellar system was formulated to dissolve tetracycline hydrochloride and water in ethyl formate with the aid of cetyltrimethylammonium bromide. The resultant micellar solution was used to dissolve 0.3 to 0.75 g of PLGA, and microspheres were prepared following a modified solvent quenching technique. As a control experiment, the drug was encapsulated into PLGA microspheres via a conventional methylene chloride-based emulsion procedure. The microspheres were then characterized with regard to drug loading efficiency, their size distribution and morphology. The reverse micellar procedure led to the formation of free-flowing, spherical microspheres with the size mode of 88 microm. When PLGA microspheres were prepared following the conventional methylene chloride-based procedure, most of tetracycline hydrochloride leached to the aqueous external phase: A maximal loading efficiency observed our experimental conditions was below 5%. Their surfaces had numerous pores, while their internal architecture was honey-combed. In sharp contrast, the new reverse micellar encapsulation technique permitted the attainment of a maximal loading efficiency of 63.19 +/- 0.64%. Also, the microspheres had smooth and pore-free surfaces, and hollow cavities were absent from their internal matrices. The results of this study demonstrated that PLGA microspheres could be successfully prepared following the new reverse micellar encapsulation technique. PMID- 15832829 TI - Determinants of unprotected sex among HIV-positive patients in South Africa. AB - This study examined the prevalence of unprotected sex, other sexual risk behaviours, and factors associated with unprotected sex among men and women recently diagnosed with HIV in South Africa. One hundred and forty-nine outpatients (44 males and 105 females) were assessed, of whom 101 were sexually active at least 6 months prior to study entry. Subjects were asked about sexual risk behaviours with reference to their most recent sexual encounter. Logistic regression analysis was employed to determine the predictors of condom use, with independent variables selected from five general categories: (1) sociodemographic characteristics; (2) situational characteristics regarding sexual intercourse (i.e. alcohol or drugs used before intercourse); (3) clinical diagnoses; (4) negative life events; and (5) coping styles. Fifty-five patients (19 males and 36 females), representing 54.4% of those sexually active in the 6 months preceding the study, had not used a condom during the most recent intercourse. Compared with those who used condoms, participants who did not significantly reported shorter duration of HIV infection (t = -2.7, p < 0.001), have a current partner (chi2 = 3.98, p = 0.005), and lack knowledge of their partner's HIV status (chi2 = 4.78, p = 0.004). Also they were significantly more likely to engage in denial (t = 3.2, p < 0.002) and to use substances (t = 1.98, p < 0.05) as a means of coping. Logistic regression showed that shorter duration of illness (odds ratio (OR) = 1.2, 95% confidence interval (CI) = 1.01-1.41) and coping styles characterized by denial (OR = 0.6, 95% CI = 0.45-0.96) were significantly associated with unprotected sex. These data suggest the need for interventions to further reduce sexual risk behaviours in HIV-positive patients in South Africa. PMID- 15832830 TI - Quality of life in HIV-infected individuals receiving antiretroviral therapy is related to adherence. AB - This study assesses changes in quality of life (QoL) over time among HIV-infected individuals receiving antiretroviral therapy (ART) and evaluates how this relates to ARTadherence. Prospective, longitudinal data were examined from 1050 participants in two large, randomized, multi-centre antiretroviral clinical trials. QoL was assessed by the SF-12; adherence by the Terry Beirn Community Programs for Clinical Research on AIDS Antiretroviral Medication Self-report. Participants included 20% women, 53% African Americans, 16% Latinos; mean age was 39 years; mean baseline CD4+ cell count 230 cells/mm3; 89% were ART-naive at entry. Baseline physical and mental health summary QoL scores were 45.4 and 42.9, comparable to scores reported in other advanced HIV populations. Significant improvements in mean QoL scores were seen for the group as a whole after 1 to 4 months on new ART regimens, and persisted for 12 months. Participants reporting 100% ART adherence achieved significantly higher QoL scores at 12 months compared to those with poorer adherence, particularly if 100% adherence was consistent (p < 0.001). Those with at least 80% ART adherence had smaller gains in QoL at 12 months when compared to baseline, while those with < 80% adherence had worsening of QoL. In this analysis, ART adherence was associated with improved QoL, particularly if adherence was sustained. PMID- 15832831 TI - Contexts for last occasions of unprotected anal intercourse among HIV-negative gay men in Sydney: the health in men cohort. AB - The objective of the paper was to compare encounters involving unprotected anal intercourse (UAI) and protected anal intercourse (PAI) among HIV-negative gay men in Sydney. Data were from those completing baseline face-to-face interviews to end June 2003 for the Health in Men open cohort of HIV-negative gay men in Sydney. The 1,148 participants ranged in age from 18 to 75 years (median = 36). Three hundred and fifty-two (30.7%) reported an occasion of UAI with a casual partner in the previous 6 months and 531 (46.3%) reported an occasion of UAI with a regular partner in that same time. The men's most recent sexual contact with a casual partner involving UAI was distinguished from those involving PAI by a greater likelihood for both partners to disclose HIV serostatus (p = 0.006) and by respondents being more inclined to restrict themselves to the insertive position or to practise withdrawal during occasions involving any UAI than when a condom was used (p = 0.003 and p = 0.001 respectively). Neither location nor recreational drug use differentiated men's most recent sexual contacts involving UAI from those involving PAI. The decision by HIV-negative gay men to use condoms during sexual encounters with either regular or casual partners is guided more by HIV serostatus and risk reduction strategies than by other factors. PMID- 15832832 TI - Sexual risk behaviour, viral load, and perceptions of HIV transmission among homosexually active Latino men: an exploratory study. AB - The objective of the paper was to describe awareness of HIV medications and HIV viral load, and to assess the impact of HIV medications (including highly active antiretroviral therapy) and notions of viral load on sexual risk practices. This was an exploratory cross-sectional study of a non-random sample of 395 homosexually active Latino men in New York City. An anonymous self-administered questionnaire was used focusing on perceptions about HIV/AIDS, HIV treatment medications and viral load, risk perceptions, HIV status, and sexual partners and practices in the past 6 months. HIV-positive participants taking HIV medications, those who knew their viral loads, and those who had undetectable viral loads did not report significantly different frequencies of high-risk (receptive or insertive) unprotected anal intercourse (UAI) in comparison to other HIV-positive individuals in the sample. Perceptions about HIV status (assuming same status) and viral load (penetrating partner or letting partner penetrate one) were related to significantly higher high-risk UAI in the past 6 months. HIV prevention strategies for homosexually active Latino men should not only address the basic issue of assuming similar HIV status but also need to examine misconstructions of the transmissibility of HIV as a result of undetectable viral load. PMID- 15832833 TI - HIV vaccine efficacy trial participation: men who have sex with men's experiences of risk reduction counselling and perceptions of risk behaviour change. AB - Qualitative interviews were conducted with 35 men who have sex with men, enrolled in the world's first phase III HIV vaccine efficacy trial at five US sites, regarding their risk reduction counselling experiences and their perceptions of its impact on risk behaviour. Respondents ranged in age from 20 to 58 years and were predominately white (71.4%) in racial/ethnic origin. Systematic qualitative analysis revealed that a positive counselling experience meant having good rapport with clinic staff. Differences in attitudes toward counselling were related to either a personal approach of balancing an enjoyable sex life with safe sex behaviours (balancing risks) or accepting the consequences of risky sexual behaviour rather than making changes (risk homeostasis). Respondents seeking to balance risks indicated that they saw themselves engaging in safer sexual behaviour almost twice as often as in riskier behaviours. They perceived counselling and behavioural risk assessments to help increase their awareness of personal risk-taking behaviours. Conversely, those with a risk homeostasis approach reported that they had established sexual boundaries prior to trial participation that had thus far proven to be effective in avoiding HIV infection, and that they were comfortable with the level of risk taken. Thus, risk reduction counselling had little to no influence on their sexual practices. Some of these men also indicated that while they had not found the risk reduction information imparted to them by clinic staff to be novel, counselling was beneficial in reinforcing their HIV/AIDS and safe sex knowledge base. PMID- 15832834 TI - Identifying, recruiting, and assessing social networks at high risk for HIV/AIDS: methodology, practice, and a case study in St Petersburg, Russia. AB - Population segments at highest risk for HIV are often hidden, marginalized, and hard to reach by conventional prevention programmes. This pattern is especially true in Central and Eastern Europe, where major HIV epidemics have recently appeared, where population members do not perceive themselves as belonging to a community, and where there is little precedence for strong community-based organization service programmes. In these circumstances, naturally existing intact social networks still can be targeted by prevention programmes. HIV prevention interventions undertaken with at-risk social networks can establish new group norms, reduce the risk behaviour of network members, and can reach 'hidden' members of a population known personally to leaders of the social networks. This article illustrates a methodology and a practical description for: (1) accessing high-risk social networks in a community population; (2) identifying and enumerating the membership of the social networks; (3) identifying the social leadership of the networks; and (4) establishing the HIV risk behaviour levels of the recruited networks. To illustrate how social network methods can be applied in the field, the article provides case study reports of HIV prevention fieldwork practice targeting high-risk networks of young men who have sex with men and young heterosexual adults in St Petersburg, Russia. Although there is an extensive conceptual literature on the influence of social networks on risk behaviour, this article describes specific and practical techniques that can be in the development of approaches for social network-based interventions. PMID- 15832835 TI - Mother-to-child transmission of HIV infection in Romania: results from an education and prevention programme. AB - A pilot prevention of mother-to-child transmission (PMTCT) programme was implemented in Constanta County, Romania, between 2000 and 2002. The programme consisted of clinician training, routine antenatal HIV counselling and testing and the care of HIV-infected pregnant women and their infants. A total of 11,423 pregnant women (10,192 (89%) white Europeans, 862 (7.5%) Rroma, 369 (3.2%) Central Asians) were tested during the pilot, at a median of 24 weeks' gestation. Rapid HIV testing at delivery was introduced during the pilot, to supplement the antenatal testing, both of which required informed consent. Overall seroprevalence was 1.75 per 1,000 (95% confidence interval (CI) 1.07-2.70 per 1,000). HIV infection was associated with having a high-risk partner, prostitution and non-Caucasian ethnicity. Twelve infected women completed their pregnancies, of whom seven received antenatal antiretroviral therapy (ART); all neonates received prophylactic ART and five were delivered by elective caesarean section. Three infants were HIV-infected, giving a vertical transmission rate of 25% (95% CI 5.49-57.2%); all three were born to mothers not identified as infected until delivery, and who therefore received no antenatal ART. A key challenge for PMTCT in Romania will be the prompt identification of pregnant HIV infected women, to allow the optimum application of interventions. PMID- 15832836 TI - Reducing discriminatory attitudes toward people living with HIV/AIDS (PLWHA) in Hong Kong: an intervention study using an integrated knowledge-based PLWHA participation and cognitive approach. AB - The present paper describes the development and evaluation of an intervention programme aiming to reduce adolescents' discriminatory attitudes toward people living with HIV/AIDS (PLWHA). The intervention programme integrates components of 'virtual interaction' with PLWHA (watching a documentary), knowledge enhancement and a simple cognitive exercise. To evaluate its effectiveness, the programme was implemented to about 600 form 3-4 (grade 9-10) students of three secondary schools in Hong Kong. Using a structured questionnaire, the level of discriminatory attitudes toward PLWHA, knowledge about HIV/AIDS and perceptions about PLWHA, etc. were measured before and after the implementation of the programme. A notable improvement on the level of acceptance of PLWHA and knowledge about HIV/AIDS was found after the implementation of the programme. Negative perceptions about PLWHA also reduced substantially. For instance, before the programme, over one-third (35.7%) of all respondents believed that the majority of PLWHA were promiscuous; the figure dropped to 15.8% after exposure to the programme (adjusted odds ratio = 0.35, p < 0.001). Further, some gender differences were observed. Female respondents tended to be less discriminatory toward PLWHA and responded more favourably to the programme than their male counterparts. PMID- 15832837 TI - Injection prevalence and risks among male ethnic minority drug users in Northern Thailand. AB - Thailand's rate of HIV due to drug use is increasing, and nearly one-half of clients at a drug treatment centre in the north are from marginalized ethnic minority groups. We describe and compare socio-demographics, drug use, and sexual practices across ethnic minority people presenting for treatment at the Northern Drug Treatment Center, Mae Rim, Thailand. A prevalence study was conducted between February 1999 and January 2000. Trained interviewers administered a baseline questionnaire to 1,865 people admitted to the Northern Drug Dependence Treatment Center. Ethnic minority (Karen, Akha, Hmong, Lisu, and Lahu) males were included in this analysis (n = 629). Between 7% and 28% of each minority group had ever injected drugs (p = 0.002). Ninety-five per cent of men ever injecting had used heroin. Younger age, speaking Thai language, having a job with higher contact with Thais, and prior drug detoxification were significantly associated with injection in multivariate analysis, while Hmong and Lisu ethnicity were protective. Using occupation and language as proxies, contact with Thais and concomitant acculturation may be a risk for injection among this population. The inadvertent consequences of acculturation as it relates to injection and HIV risk must be addressed. PMID- 15832840 TI - Educate early, educate often. PMID- 15832838 TI - Exploring direction of causation between social support and clinical outcome for HIV-positive adults in the context of highly active antiretroviral therapy. AB - Social support and its associations with virological outcome were assessed over a 4-year period for 34 adult outpatients living with HIV/AIDS and consistently on highly active antiretroviral therapy (HAART). Clinical variables and perceived availability of social support were measured in three waves (T1, 1997; T2, 1999; T3, 2001). Cross-lagged logistic regression analyses were employed to evaluate the direction of potential causal pathways between social support and clinical status in terms of 'undetectable' viral load. Social support ratings declined moderately overall, with a more pronounced reduction that was of clinical (effect size, 0.82) and statistical significance (p = 0.000) for the subgroup (n = 11; 32%) with final 'detectable' viral load. Evidence suggested causal directionality in which cross-sectional social support and/or improvements in social support over time predicted virological outcome, with better social support associated with greater likelihood of viral load suppression to 'undetectable' level, an outcome achieved for 68% (n = 23) of the sample. In contrast, cross-sectional virological status reflected immunological outcomes but did not predict subsequent ratings of social support or changes in social support ratings. HIV positive adults consistently taking HAART appeared to experience better clinical benefit if they perceived interpersonal, informational and emotional support to be available, a finding that underscores the importance of social support in relation to treatment outcome for this population. PMID- 15832841 TI - Educating high school students about the profession of optometry. PMID- 15832842 TI - Glaucoma without cupping. PMID- 15832843 TI - The relationship between central corneal thickness-adjusted intraocular pressure and glaucomatous visual-field loss. AB - BACKGROUND: Although measurement of central corneal thickness (CCT) is increasingly becoming an important component of glaucoma risk analysis, significant controversy exists regarding the benefit of calculating a corrected intraocular pressure (IOP) value from measured IOP and CCT data. METHODS: Three hundred forty-four male subjects were identified from a VA eye clinic with one of the following clinical diagnoses: ocular hypertension (OHT), primary open-angle glaucoma (POAG), normal tension glaucoma (NTG), and normal tension glaucoma suspect (NTGS). Using one eye per subject, multivariate logistic regression and correlational analyses were performed to determine relationships between glaucomatous visual-field loss and several glaucoma risk factors, including adjusted IOP values. RESULTS: Multivariate logistic regression analysis did not identify CCT-adjusted IOP values as independent risk factors for development of either NTG or POAG-related glaucomatous visual-field loss. CCT, however, was found to be strongly associated with both NTG and POAG-related visual-field loss. Correlational analysis revealed a weak correlation between Ehlers-adjusted pre treatment IOP and severity of POAG-related visual-field loss, but no other adjusted IOP values significantly correlated with severity of visual-field loss in either POAG or NTG. CONCLUSIONS: Our results suggest that adjusted IOP, as calculated using current algorithms, is not useful within glaucoma risk analysis, since adjusted IOP was unable to predict either presence or severity of glaucomatous visual-field loss in this study. CCT, conversely, was found to be a robust and independent predictor of glaucomatous visual-field loss. These findings, while supporting routine CCT measurements for all glaucoma suspects, do not support routine clinical computation of adjusted IOP values using current algorithms. PMID- 15832844 TI - Bilateral disk edema with unilateral macular serous fluid secondary to neurocysticercosis. AB - PURPOSE: The purpose of this article is to document a case of neurocysticercosis that manifested clinically with bilateral disk edema and serous fluid accumulation in the macula of the left eye. We also describe the recovery of visual-field loss (O.D.) and diminution of bilateral disk edema following anti helminthic treatment. CASE REPORT: A 41-year-old woman reporting headaches came to us with bilateral disk edema and co-existing serous macular fluid in the left eye. RESULTS: Magnetic resonance imaging (MRI) revealed the presence of large multi-septated cysts that exhibited peripheral enhancement, with minimal surrounding edema at the posterior left temporal lobe. Scattered punctate lesions, suggesting areas of calcification, were also observed within both temporal lobes. The optic chiasm appeared compressed. This presentation was considered characteristic of neurocysticercosis and the patient was prescribed a regimen of 200mg albendazole b.i.d. The patient responded well to the treatment, with progressive resolution of the bilateral disk edema and macular fluid. CONCLUSION: Although rare, cases of cysticercosis and neurocysticercosis may still be encountered in industrialized nations, where the parasite has been almost eradicated. Ocular signs of disk edema and macular serous fluid secondary to neurocysticercosis may correspond to optic nerve, parenchymal, or extraparenchymal disease. PMID- 15832845 TI - Orbital myositis following streptococcal pharyngitis in a pediatric patient. AB - BACKGROUND: Orbital myositis is a relatively rare ocular inflammatory disease. It is currently classified as an idiopathic orbital inflammatory disease, but has been associated with ocular and systemic disorders, including scleritis, rheumatoid arthritis, Crohn's disease, and systemic lupus erythematosis. Orbital myositis has also been associated with infectious disease such as streptococcal pharyngitis, viral upper respiratory infection, and Borrelia burgdorferi infection. CASE REPORT AND DISCUSSION: This report documents treatment and imaging of a 13-year-old boy who had an episode of right-sided orbital myositis following streptococcal pharyngitis with anti-streptolysin-o (ASO) titer elevation to 1,188 IU/ml. Clinically, this patient demonstrated acute onset of marked restriction and mild under-action of the lateral rectus muscle of the right eye. Thickening of the lateral and medial recti was demonstrated on computed tomography. The mild orbital myositis was treated with oral nonsteroidal anti-inflammatory drug therapy. The nonconcomitant strabismus resolved approximately 7 weeks after initial presentation. General orbital inflammatory disease management and the association of upper respiratory tract infection with orbital myositis are discussed. CONCLUSION: Orbital myositis is a distinct clinical entity that can occur after streptococcal upper respiratory infection, consisting of pain on ocular movement, localized swelling, restrictive strabismus, and inflammatory signs of the involved muscle(s). PMID- 15832846 TI - A commonsense approach to the HIPAA security regulation. PMID- 15832847 TI - Technical safe guards for the optometric practice. PMID- 15832848 TI - Investing in a practice will keep it ahead of the curve. PMID- 15832849 TI - Keeping patient divorces out of your optometry practice. PMID- 15832850 TI - The power to choose. PMID- 15832851 TI - Complementarity of IUHPE and WHO conferences on health promotion. PMID- 15832852 TI - The IUHPE's response to the Tsunami disaster. PMID- 15832853 TI - Harnessing support for prioritised Cochrane reviews in health promotion and public health--update from the Cochrane Health Promotion and Public Health Field. PMID- 15832854 TI - Global health promotion and health education initiatives: an update and what's in store for the third programme year of the IUHPE/CDC co-operation. PMID- 15832855 TI - 2nd International Conference on Local and Regional Health Programmes: strengthening the integration of promotion and prevention in health systems. Quebec city, Quebec, Canada--October 12-15, 2004. PMID- 15832856 TI - [Investigation on the action and construction of power in health education]. PMID- 15832857 TI - [Potential and limitations of the health sector networks and wellbeing: the example of the red Quebequense for cities and healthy municipalities (RQMS)]. PMID- 15832858 TI - Assessment of tongue weakness and fatigue. AB - Assessment of nonspeech tongue function is common in speech-language pathology. This paper reviews techniques used to determine tongue strength and endurance, and describes a constant-effort task. These techniques are intended to reveal and quantify the presence of weakness or fatigue of the tongue. The consequences of performing these tasks with and without a bite block, used to fix jaw position, are considered. Whether nonspeech tongue impairment is associated with speech dysfunction in Parkinson's disease is another topic of interest. Past studies indicated reduced tongue strength and endurance in Parkinson's disease, but these measures did not correlate with speech measures. It was hypothesized that weakness and fatigue need to be impaired to a "critical" level before speech is perceptibly affected. To examine whether experimentally induced tongue fatigue affects speech, normal speakers performed prolonged strenuous tongue exercise. Speech deteriorated following these exercises. A new investigation examines whether 1 hour of speech-like tongue exercise (rapid syllable repetitions) affects dysarthric speech. Preliminary data from 6 participants with Parkinson's disease, 1 person with bulbar ALS, and 6 neurologically normal control subjects indicate that sentences sound more precise but less natural after the exercises. Surprisingly, results did not differ significantly between the groups. Continued collection of data and refinement of tasks will contribute to our understanding of the potential relationships between weakness, fatigue, and speech. PMID- 15832859 TI - The role of animal models in understanding feeding behavior in infants. AB - The common evolutionary history humans share with mammals provides us with a solid basis for understanding normal oropharyngeal anatomy and functions. Physiologically, feeding is a cycle of neurophysiologic activity, where sensory input travels to the CNS which sends motor signals out to the periphery. Research with animal models is valuable because it is possible to disrupt this cycle, and develop predictive models on the causal basis of deviation from normal. Based on work with animal models, normal mammalian infant feeding behavior consists of the tongue functioning as a pump. First, the tongue assists in acquisition of milk from the nipple into the oral cavity, and then it pumps milk from the oral cavity into the valleculae prior to the pharyngeal swallow. Starting with this basic model, feeding in infant pigs was manipulated to determine the impact of variation in sensory input on behavioral output. One set of experiments suggested that chemo- or liquid sensation, in the form of milk is necessary to elicit continuing rhythmic activity. However, the rates of rhythmic suckling are intrinsic to an animal, and variation in rate cannot be entrained. Another set showed that initiation of the swallow does not purely depend on the volume of milk delivered, but also on the sensory stimulation at the mouth. These results support the idea that feeding behavior involves complex sensory integration. PMID- 15832860 TI - Lingual frenulum: classification and speech interference. AB - PURPOSE: To propose a classification of the different lingual frenulum and to relate them to speech disorders. METHODS: We evaluated 1402 patients' frenulum with an age range of 5 years 8 months to 62 years 10 months between 1978 and 2002. Pictures were taken of the altered frenulum. Measures of maximal mouth opening, with and without tongue suction, were taken with a sliding caliper. Speech samples were also taken. Frenulum were then classified as normal; short; with anterior insertion, and short with anterior insertion. RESULTS: From the 1402 patients evaluated, 127 (9%) presented with an altered frenulum insertion. For this study we considered only those with short or with anterior insertion. For those who had an altered frenulum, 62 (48.81%) presented with speech disorders. The more frequent speech disorders were: omission and substitution of /r/; {R}, and consonant clusters with /r/, and of /s/ and /z/. Frontal and lateral lisps also occurred. The frenulum of 21 patients was classified as short and of these, 12 patients (57%) presented with speech disorders. Of the 106 patients with anterior insertion, 50 (47.2%) presented with a speech disorder. After statistical analyses the relation between altered frenulum and speech disorders was considered significant with p<0.001. CONCLUSION: The lingual frenulum was classified as normal, short and with anterior insertion. An altered frenulum may predispose the individual to exhibit an accompanying speech disorder. PMID- 15832861 TI - Evaluation of a new concept of myofunctional therapy in children. AB - This prospective study was designed to evaluate a new concept of myofunctional therapy in comparison with conventional myofunctional therapy. 45 children aged three to sixteen years in need of myofunctional therapy were randomly divided into two groups: 19 children were referred to myofunctional therapy in private practices in Hamburg and served as controls. The remaining 26 children were treated with face former therapy at the Department of Orthodontics by a medical assistant specializing in myofunctional therapy. The overall observation time was six months. Every three months an overall clinical assessment was performed at the Department of Orthodontics by a speech pathologist and an orthodontist, who documented the clinical situation. The clinical examination included measurement of lip strength, palatography to document the swallowing pattern, logopedic diagnosis, and an orthodontic examination with reference to a standardized diagnostic sheet. In all children's orofacial function could be improved. Children treated with the Face Former showed a statistically significantly improvement in palatal tongue position during swallowing. They achieved stronger lip pressure within a shorter time than children who did not use the Face Former. However, at the end of the observation time there was no statistically significant difference in lip strength between the two groups. Habitual mouth closure was also achieved within a shorter time for children treated with the FaceFormer than children with myofunctional therapy. Face Former therapy seems to offer a good alternative to conventional myofunctional therapy. Longitudinal studies will follow to judge whether the established orofacial balance could be stabilized, i.e. the established physiological orofacial function becomes automatic. PMID- 15832862 TI - Distribution of causes and treatments of dysphagia at dysphasia/dysphagia rehabilitation clinic of Showa University Dental Hospital: 1999-2002. AB - The purpose of this study was to investigate the distribution of causes of dysphagia and the types of treatments being provided at the Dysphasia/Dysphagia Rehabilitation Clinic of Showa University Dental Hospital. The subjects included 173 dysphagic patients. Fifty-three percent of patients ranged 0-12 age, and 24% of them were over 60 years old. The survey results showed that diseases of the central nervous system, such as cerebral palsy (CP), was a major diagnosis in young patients, and cerebrovascular accident (CVA) was a predominant cause in adult and elderly patients. Diagnosis distribution showed that swallowing dysfunction was the most frequent diagnosis for patients on their first visit when compared to other feeding dysfunctions including malfunction of lips, tongue, and mastication. Almost 40 % of patients still continue to have rehabilitation for their dysphagic symptoms at the end of March 2002. This suggests that dysphagia rehabilitation is needed for a long time for many disabled individuals. PMID- 15832863 TI - Early lingual frenectomy assisted by CO2 laser helps prevention and treatment of functional alterations caused by ankyloglossia. AB - Incorrectly produced speech sounds, the presence of dentofacial alterations and acquired functional adaptations may be due to a short and inadequate lingual frenum. When frenectomy is indicated, it should be performed as early as possible to prevent functional alterations. This study presents a literature review on correct lingual positioning in relation to orthodontic and phonetic function as well as an assessment of 15 patients who underwent frenectomy utilizing the carbon dioxide laser. The results demonstrated that this technique is safe, effective and perfect for use in young children and can be performed in an outpatient unit. PMID- 15832864 TI - A morphological comparison of cervical spondylotic myelopathy: MRI and dissection findings. AB - The number of people with functional disabilities has been increasing with the rapid changes of age structure in the overall population. One of the major causes of disturbances in daily activities is cervical spondylotic myelopathy (CSM). The transverse area and sagittal diameter of the spinal cord measured by MRI is reported to correlate with the clinical manifestations of CSM, the duration of the disease, and the speed of recovery after surgery in patients with CSM. The purpose of this research is to determine the morphological characteristics of CSM as seen in MRI findings including the spinal cord sagittal diameter, transverse diameter, transverse area and flatness ratio. Twenty-eight of several patients with CSM were treated conservatively after carrying out measurements by MRI. In addition, anatomical studies were carried out on the spinal cords after anatomical dissection of the vertebral column in seven cadavers with CSM. These results, when compared with the morphological analysis of the cervical spinal cord, show that there is a correlation between the transverse areas at C4, C5 and C6 levels, as those at every level of the cervical transverse areas of tissue sections in the dissection cadavers were 10-18% smaller than those in the MRI patients. These results should be taken into account for the treatment of CSM patients. PMID- 15832865 TI - Aging process of myelinated nerve fibers in the human Lissauer tract. AB - We calculated numbers and axonal areas of myelinated nerve fibers in the Lissauer tract of the human lumbar spinal cord (L1) from the viewpoint of the aging process. We examined 20 human spinal cords from 13 males and 7 females, age ranging from 41 to 88 years old. We found that, although the number of nerve fibers showed no significant change in relation to the age of the subject, the axonal area of myelinated nerve fiber in the Lissauer tract did decrease with age. PMID- 15832866 TI - Morphometric difference in the human maxillary nerve fibers between dentulous and edentulous jaw subjects. AB - This study was conducted to quantify the change in the number and size of myelinated nerve fibers of the maxillary nerve with tooth loss in humans. We carried out a morphometric analysis to compare the number and size of myelinated nerve fibers in the human maxillary nerve between four dentulous and four edentulous jaw cases. Our results indicated that the number of axons decreased by approximately 13,000 with tooth loss. The average size of axons remained unchanged, but there was a change in the fiber size distribution, namely the loss of a large number of small-sized axons was accompanied by the total disappearance of small number of large-sized axons. PMID- 15832867 TI - Comparative anatomical study of the kidney position in amniotes using the origin of the renal artery as a landmark. AB - The anatomical relationship between the kidney position and its arterial supply was investigated in 21 mammals, 1 bird, and 3 reptiles (n = 1 for each species) and in 43 human cadavers. The following observations were made. (1) Although the right kidney was located caudal to the left kidney in 29 out of 43 human cadavers (67.4%), the origin of the right renal artery from the aorta was located cranial to the origin of the left renal artery in 36 human cadavers (83.7%). Therefore, the relative positions of the kidneys do not correspond with the relative origins of the renal arteries in humans. (2) Among the mammals that were examined, the position of the kidney and the branching level of the renal artery on the right side were usually cranial to those on the left side. (3) In the bird and most reptiles that were examined, kidneys were typically located in the pelvic region and were supplied by segmental arterial branches. These results suggest that the right kidney and its arterial supply are generally located cranial to the left kidney in phylogeny of mammals. While the presence of a human accessory renal artery in 9 out of 86 sides (10.5%) and a cranial origin of the left renal artery relative to the right renal artery in 7 out of 43 cadavers (16.3%), shows some variation in the arterial supply to the kidneys, the origin of the renal arteries can generally be used as phylogenetic landmarks indicating the relative positions of the kidneys. Hence, from an ontological perspective, the human right kidney may be initially situated cranial to the left kidney during the early stages of development. Thereafter, the human right kidney may shift downwards secondary. PMID- 15832868 TI - Prevention of vascular injury to the spinal cord incidence of a great posterior radicular artery. AB - This study investigated in 165 cadavers for the presence and morphology of a great posterior radicular artery (GPRA). The spinal cords were examined for the presence of a GPRA macroscopically and by stereomicroscopy, and the spinal segmental level where the GPRA was located and left-right differences were investigated. A GPRA was found to be present in 10 (6.1%) of the 165 cadavers, and a solitary unilateral GPRA was observed on the left or right side of the spinal cord in each of them. While no differences in laterality were identified, the GPRA was most often seen at the level of the spinal segment T11. The mean outer diameter of the GPRA was 0.43 mm. No previous studies have described the GPRA in a Japanese population. In the 10 cadavers in which a GPRA found in this study its most common location was at the level of spinal segment Th11. There was a difference of more than 2 spinal segmental levels between the location of the artery in our study and in a European population, suggesting that the spinal segmental level at which the GPRA is located is 1 or 2 levels higher in the Japanese population. PMID- 15832869 TI - Life satisfaction and disability after severe traumatic brain injury. AB - OBJECTIVE: To assess the relationships between life satisfaction and disability after a severe traumatic brain injury (TBI). DESIGN: Cross-sectional study, including 75 patients 2 years or more after a severe TBI. METHODS: Life satisfaction was assessed with the Subjective Quality of Life Profile. Impairments, activities and participation were assessed with standardized tests. RESULTS: The satisfaction profile was flat, i.e. the majority of items obtained mean satisfaction scores close to 0, suggesting that participants felt indifferent to these items or in other words that they were neither satisfied nor unsatisfied. Patients were on average slightly dissatisfied with their cognitive functions, physical abilities and self-esteem. A factor analysis revealed three underlying factors. The main finding was that the relationships between life satisfaction and disability were not linear: the lowest satisfaction scores were reported by participants with moderate disability rated by the Glasgow Outcome Scale, while individuals with severe disability did not significantly differ from the good recovery group. CONCLUSION: Life satisfaction is not linearly related to disability after severe TBI. PMID- 15832870 TI - Neuropsychological outcome and community re-integration following traumatic brain injury: the impact of frontal and non-frontal lesions. AB - PRIMARY OBJECTIVE: To examine the relationship between cortical lesion location and brain injury outcome. It was hypothesized that focal frontal lesions after traumatic brain injury (TBI) would result in decreased executive and memory functioning and poor community participation outcome. RESEARCH DESIGN: Three quasi-experimental, prospective studies employed a total of 643 patients with focal frontal, fronto-temporal, non-frontal or no lesions in CT scans. METHODS AND PROCEDURES: CT scan analysis, neuropsychological assessment, the Neurobehavioural Functioning Inventory (NFI), the Community Integration Questionnaire (CIQ). MAIN RESULTS: In study 1, frontal and fronto-temporal groups performed worse in executive functioning and better in constructional ability. Study 2 found no differences in neuropsychological and community re-integration measures at 1-year follow-up. Study 3 found comparable neuropsychological test score improvement across groups over 1 year. CONCLUSIONS: Results are consistent with previous findings and document the potential for test score improvement with rehabilitation and suggest that lesion location needs to be considered when individual rehabilitation plans are being implemented in the post-acute stage of TBI. PMID- 15832871 TI - Effect of traumatic subarachnoid haemorrhage on neuropsychological profiles and vocational outcome following moderate or severe traumatic brain injury. AB - OBJECTIVE: Traumatic subarachnoid haemorrhage (tSAH) frequently occurs in moderate or severe traumatic brain injury (TBI) and is related to worse outcome at time of discharge from the acute hospitalization. The current study compared neuropsychological impairment and vocational outcome at 1-year post-injury in patients with and without tSAH. DESIGN: Acute injury, neuroradiological, neuropsychological and vocational data were collected for 100 patients admitted for neurorehabilitation following TBI. RESULTS: Patients with tSAH had significantly worse vocational outcome than patients without tSAH. On neuropsychological measures, patients with tSAH generally performed worse than patients without tSAH across most neurocognitive domains. However, differences in neuropsychological test performance between patients with and without tSAH reached statistical significance on measures of visuospatial processing, verbal reasoning and mood only. CONCLUSION: The presence of tSAH appears to be associated with worse vocational outcome in survivors of moderate or severe TBI. As such, the presence of tSAH appears to have predictive value with respect to outcome following TBI. PMID- 15832872 TI - Effect of an integrated reality orientation programme in acute care on post traumatic amnesia in patients with traumatic brain injury. AB - PRIMARY OBJECTIVE: To assess the effect of an integrated reality orientation programme (North Star Project) in acute care on the duration of post-traumatic amnesia (PTA) of patients suffering from traumatic brain injury (TBI). It was hypothesized that those patients having participated in this programme would show a shorter PTA than those patients that did not. METHOD: Duration of PTA for 12 patients with TBI who were part of the North Star Project was compared with that of 26 patients matched for initial GCS and age for whom this approach was not available (control group). RESULTS: Length of PTA was shorter by 5 days for the North Star patients. This result was not statistically significant (p = 0.19) but is clinically relevant. No between-group difference was found for Glasgow Coma Scale. CONCLUSIONS: Consequences of shorter PTA would include increased and more appropriate patient interaction and earlier transfer to rehabilitation. The North Star project also allows the family to become more actively involved in the treatment of their loved one. Further study is required to evaluate this intervention strategy to improve orientation for the patient with TBI. PMID- 15832873 TI - Prospective and declarative memory problems following moderate and severe traumatic brain injury. AB - PRIMARY OBJECTIVE: To examine prospective and declarative memory problems following moderate and severe traumatic brain injury (TBI) and the relationship between prospective memory (PM) and declarative memory and PM and other cognitive functions. RESEARCH DESIGN: The performance of persons who suffered a TBI (n = 25) was compared with that of a demographically matched control group (n = 25). METHODS AND PROCEDURES: Measures of time- and event-based PM, visual and verbal declarative memory, attention and executive functioning were administered to both groups. MAIN OUTCOME AND RESULTS: The group with a TBI performed more poorly on event- and time-based PM, verbal declarative memory, certain aspects of attention and executive functioning. The correlations between the measures of PM, declarative memory and the other cognitive tests were all non-significant. CONCLUSIONS: Problems with declarative memory, attention, and executive functioning do not adequately account for poorer PM performance following a TBI, suggesting that PM should also be assessed following TBI. PMID- 15832874 TI - Head injury and family carers: a pilot study to investigate an innovative community-based educational programme for family carers and patients. AB - PRIMARY OBJECTIVE: The incidence of head injury is increasing among younger people with more family members undertaking their life-long care. Many research studies have highlighted the emotional well-being of such family carers and their unmet needs, however only a few consider the formal help provided for carers. Using a longitudinal, mixed variable, within and between-subject design, this pilot study evaluated the impact of an educational programme for family carers and their head-injured relatives in reducing carer and patient psychological distress and improving their coping ability. MAIN OUTCOMES AND RESULTS: The study comprised experimental and control samples each with carer and patient groups. The experimental sample had eight sessions of educational input. All groups were assessed pre- and post-intervention and at 3 months follow-up. The patient sample was further assessed using cognitive measures. There was evidence of reduction in psychological distress in the experimental carer group following the educational input, but these results were not statistically significant. However, the experimental patient population at follow-up assessment showed statistically significant improvements. CONCLUSIONS: A larger scale multi-centre study with a longer follow-up period of assessment is required for the generalization of findings. The pilot study identifies points for consideration in a potential main study. PMID- 15832875 TI - Reduction of affective lability and alcohol use following traumatic brain injury: a clinical pilot study of anti-convulsant medications. AB - OBJECTIVE: A large and under-recognized sub-set of patients suffer both traumatic brain injury (TBI) and alcohol dependence (ADep). This group appears to use alcohol to self-treat affective and anxiety lability following TBI, resulting in new ADep or worsened prior ADep. This study hypothesized that treatment of such patients with mood-stabilizing medications would relieve post-TBI emotional dysregulation and facilitate reduction in alcohol use. DESIGN: This study reported retrospective medical record data from outpatients in the Substance Abuse Treatment Programme who were treated for labile mood. Medications followed clinical indication and were given in non-blind fashion. METHOD: Subjects included 18 patients who (1) complained of debilitating affective lability following TBI, (2) described drinking alcohol to ease lability symptoms, (3) met DSM-IV criteria for current ADep and (4) were treated with a mood stabilizing medication. RESULTS: During 6 weeks of treatment, 16 (89%) achieved abstinence from alcohol. All but two (14/16 or 88%) also showed improvement in their affective and anxiety symptoms. CONCLUSIONS: These preliminary data are limited by the retrospective collection, clinical impression and non-blinded trial. Nonetheless, the results suggest further investigation of anti-convulsants as potentially useful agents in co-morbid emotional lability and ADep following TBI. PMID- 15832876 TI - Men who have sex with men (MSM) in public sex environments (Pses): a systematic review of quantitative literature. AB - We systematically review quantitative research relating to the sexual behaviours of MSM in PSEs. We examine the methodological rigour of these studies to determine an appropriate framework for future PSE-based research and quantify sexual behavioural trends therein. Medline, BIDS, Web of Science and recent HIV/AIDS conferences were searched according to a systematic inclusion criteria. Nine papers were included for review. Recruitment of participants' outwith PSE settings, and low response rates (6%) of participants contacted in situ, question the validity and generalizability of current evidence. Most PSE users were gay or bisexually identified and half of men in the gay community reported recent PSE use. Around 10% of men reported casual status-unknown/serodiscordant unprotected anal intercourse (UAI) within PSEs. HIV testing rates amongst PSE users were similar to the wider gay community, though the proportion of men who tested positive was twice as high. Rates of casual UAI suggest that PSEs represent important sites for HIV prevention. However, since extant evidence is scant and methodologically flawed, further research is urgent. Such work must recruit participants in situ, and obtain satisfactory response rates, to be generalizable to the wider population of men who cruise. PMID- 15832877 TI - Evaluation of a sexual health approach to reducing HIV/STD risk in the transgender community. AB - Despite reports of high prevalence of human immunodeficiency virus (HIV) and other sexually transmitted diseases (STDs) among the transgender community, very little prevention education has targeted this population. To fill this gap, we developed and evaluated a transgender-specific intervention, All Gender Health, which incorporates prevention strategies into comprehensive sexuality education. Transgender participants (N=181) attended the two-day seminar in community-based venues. The curriculum was delivered via lectures, panel discussions, videos, music, exercises and small group discussions. Attitudes toward condom use, safer sex self-efficacy and sexual risk behaviour were evaluated before participation in the intervention (pre-test), immediately after participation (post-test) and at three-month follow-up. Compared to pre-test values, significant improvements were seen in attitudes toward condom use and in safer sex self-efficacy at post test, and in attitudes toward condom use, increased monogamy and decreased sexual risk behaviour at three-month follow-up. Pre-test data identified unprotected anal, vaginal and oral sex as the most commonly reported risk behaviours. Many respondents also indicated problems with social discrimination, depression, suicidal ideation and sexual functioning. Future interventions should address these risk co-factors. Alternative interventions need to be developed to target those who, as a result of social marginalization, are less likely to be reached with an intensive seminar-based intervention. PMID- 15832878 TI - The acceptability of male circumcision as an HIV intervention among a rural Zulu population, Kwazulu-Natal, South Africa. AB - Epidemiological and biological studies provide compelling evidence for the protective effect of male circumcision against the acquisition of HIV. Three randomized controlled trials are currently underway to assess the impact of male circumcision as an HIV intervention in traditionally non-circumcising areas with high levels of heterosexually-transmitted infection. This study explores the acceptability of male circumcision among the rural Zulu around Hlabisa and Mtubatuba, KwaZulu-Natal, South Africa. A cross-sectional convenience sample of 100 men and 44 women was surveyed, and two male focus groups held, to ascertain circumcision preferences within the population. Four in-depth interviews with service providers assessed the feasibility of promoting male circumcision. Fifty one per cent of uncircumcised men and 68% of women favoured male circumcision of themselves or their partners; while 50% of men and 73% of women would circumcise their sons. For men, the main predictors of circumcision preference pertained to beliefs surrounding sexual pain and pleasure; for women, knowledge about the relationship between male circumcision status and STI acquisition was the key indicator for circumcision preference. Among both sexes the main barrier to circumcision was fear of pain and death. The greatest logistical barrier was that circumcision can presently only be carried out by trained hospital doctors. PMID- 15832879 TI - A randomized controlled trial to test an HIV-prevention intervention for Latino gay and bisexual men: lessons learned. AB - At the time of this writing, no randomized controlled trial (RCT) of an intervention to reduce unsafe sex among Latino gay and bisexual men (LGBM) had been published. We report the results of an RCT conducted in New York City in which 180 LGBM were assigned either to an intervention developed specifically for this population or to a wait-list control group. The intervention was based on empowerment theory and used factors identified in prior research as determinants of unsafe sex. By eligibility criteria, all men had engaged in unprotected anal intercourse (UAI) within two months of the baseline assessment. At first (two months) and second (six months later) follow-up assessments, approximately half of the men in the experimental group reported no UAI. Yet, a similar proportion of the control group also reported no UAI. Baseline data indicate that although the men had been the subject of social oppression and sexual prejudice (homophobia), they did not feel disempowered, externally controlled or fatalistic, and they reported self-efficacy and intentionality to enact safer sex. Lessons learned are discussed, as well as notes of caution for future research employing a similar conceptual framework. PMID- 15832880 TI - Using a web-based survey to assess correlates of intention towards HIV testing among never-been-tested but sexually experienced college students. AB - The purpose of this study is to identify influential psychosocial factors predicting testing intention among college students who have never been tested but are sexually experienced. A web-based survey was conducted during the spring semester in 2003 in a major Southeastern university (N = 440). Results showed only 7.7% of the students expressed intention despite having been sexually experienced. Multiple logistic regression models showed that the combined five psychosocial factors examined (perceived pros, cons, risk, self-efficacy and availability/accessibility related to testing) significantly predicted testing intention (p < 0.001). Two factors, perceived pros (OR = 1.641) and perceived risk (OR = 1.357), showed significant coefficients. The reduced model with only these two predictors showed increased correct classification of students in terms of their testing intention (from 91.8 to 93.4%). Results suggested that educational and prevention programmes aimed at encouraging testing should address these important testing-related psychosocial constructs to maximize programme impacts. Future studies need to further investigate the role of other factors in affecting testing intention or related behaviours among other sub-groups of students. PMID- 15832881 TI - HIV prevention heuristics and condom use among African-Americans at risk for HIV. AB - The goal of this study was to improve understanding of whether incorrect HIV/AIDS heuristics and characteristics-based risk theories are barriers to HIV prevention among young African-Americans at increased risk for HIV. We explored: (1) the beliefs of men and women regarding disease prevention strategies, and (2) the relationship of such beliefs to safer sexual behaviours. In Phase I, semistructured individual interviews were conducted with both members of 22 heterosexual couples at increased risk for HIV/STIs. Subsequently, in Phase II, structured individual interviews were conducted with another 40 women and 40 men (not couples). Participants in Phase I reported use of condoms and monogamy as major strategies for disease prevention. The beliefs that were endorsed by the largest percentage of Phase II participants were related to the 'known partners are safe partners' and 'trusted partners are safe partners' heuristics. Moreover, stronger endorsement of the 'known partners' heuristic was negatively associated with measures of condom use and pregnancy prevention behaviour. Our findings highlight the need for interventions and programmes to encourage intimate partners to consistently use condoms until both members of the dyad are tested and agree to mutual monogamy. PMID- 15832882 TI - Measuring adherence to treatment of paediatric HIV/AIDS. AB - Parent, child, physician report and pill counts were used to measure adherence in paediatric HIV. Relationships to viral load were assessed. Pill counts were considered invalid. Adherence measures did not correlate with one another. Physicians reported lower adherence than parents, but parent and physician report correlated with viral load. The clinical and research utility of the various measures are discussed. PMID- 15832883 TI - Stress, social support and housing are related to health status among HIV positive persons in the deep south of the United States. AB - Self-report health status measures are consistently associated with medical outcomes and are cost-effective. Studies using such measures find that those who live in rural areas or have limited access to support experience poorer health status and poorer outcomes. A survey addressing these issues was administered to 401 HIV-positive persons throughout Alabama. Hierarchical regression models examined the relation of housing stability, stress, substance use and other variables to physical (PCS) and mental (MCS) health status. The sample was 34% female and 66% African-American. Most were receiving antiretroviral medications, and nearly 20% were in unstable housing. Age showed a significant negative relationship to health status. CD4 cell count (p < 0.01) was positively associated with PCS; perceived general stress (p < 0.02) and housing stability (p < 0.04) were negatively associated. The model accounted for 14% of the variance in PCS (p < 0.001). For MCS, general stress (p < 0.001) was negatively associated and substance use tended towards a negative association (p < 0.075). Social support (p < 0.02) was positively associated with MCS. The model accounted for nearly 31% of the MCS variance (p < 0.001). Health status among HIV-positive persons may be improved by assessing and addressing social issues such as social isolation, life stressors and housing. PMID- 15832884 TI - Access to antiretroviral therapy among HIV/AIDS patients in Khon Kaen Province, Thailand. AB - This study attempted to identify the factors associated with the access to antiretroviral therapy (ARV) among HIV/AIDS patients in Khon Kaen Province, Thailand. We collected medical and sociodemographic data from the medical charts of adult patients living in the province who received medical services at two public hospitals in the province. The study period was from December 1, 2001 to February 28, 2002. Total 593 outpatients were included in the analysis. One hundred and forty-six patients (24.6%) received ARV. A logistic regression analysis was conducted to identify the factors associated with the use of ARV. Patients who were covered by the Civil Servant Medical Benefit Scheme were significantly more likely to receive ARV than those who were covered by the Universal Coverage Scheme (UC), a publicly-funded medical insurance (OR = 12.43; 95% CI = 6.03-25.62). The results of this study indicated that there were inequalities in access to and use of ARV among HIV/AIDS patients by health insurance status. The current government announced that they would include ARV in the benefits package of UC. It would be important to monitor how this policy will improve the access to ARV among HIV/AIDS patients. PMID- 15832885 TI - Patients' perception of information about HAART: impact on treatment decisions. AB - The aim of this study was to profile patients' satisfaction with information they have received about HAART in relation to treatment uptake. As part of a prospective investigation into uptake and adherence to HAART, 115 participants, comprising predominantly gay men, completed validated questionnaires investigating their satisfaction with information relating to practical aspects and potential problems of HAART, perceptions of information from different sources and beliefs about HAART, following a clinically recommended treatment offer. There was a wide range of total satisfaction scores indicating variation in patients' information requirements. Those who declined HAART were less satisfied with the information they had received than those who accepted the treatment recommendation (p < 0.05). Lower levels of satisfaction were associated with stronger concerns about the potential adverse effects of HAART (p < 0.05). Furthermore, concerns about HAART were related to uptake of HAART with those declining treatment being significantly more concerned about potential adverse effects than those who accepted the treatment offer (p < 0.001). The most helpful sources of information were specialist HIV staff: hospital consultants, pharmacists and nurses, as well as other people with HIV. This study improves our understanding of how information about HAART is perceived by people faced with treatment decisions. It emphasizes the importance of providing information tailored to meet the needs of individual patients and address their specific concerns, in order to support informed decision making. PMID- 15832886 TI - High-risk behaviour in young men attending sexually transmitted disease clinics in Pune, India. AB - The present study reports sexual risk factors associated with HIV infection among men attending two sexually transmitted disease (STD) clinics in Pune, India and compares these behaviours between young and older men. Between April 1998 and May 2000, 1872 STD patients were screened for HIV infection. Data on demographics, medical history and sexual behaviour were collected at baseline. The overall HIV prevalence was 22.2%. HIV risk was associated with being divorced or widowed, less educated, living away from the family, having multiple sexual partners and initiation of sex at an early age. The risk behaviours in younger men were different to older men. Younger men were more likely to report early age of initiation of sex, having friends, acquaintances or commercial sex workers as their regular partners, having premarital sex and bisexual orientation. Young men were more educated and reported condom use more frequently compared with the older men. Similar high HIV prevalence among younger and older men highlights the need for focused targeted interventions aimed at adolescents and young men and also appropriate interventions for older men to reduce the risk of HIV and STD acquisition. PMID- 15832887 TI - Mapping and visualizing the location HIV service providers: an exploratory spatial analysis of Toronto neighborhoods. AB - Efforts have been made to identify, reduce and ultimately eliminate health disparities, yet variation in access to health services continues to be an important concern. As with large American cities, Toronto has been particularly hard hit by the AIDS epidemic, representing 68% of Ontario's HIV diagnoses (Health Canada, 2000). The accessibility of healthcare in terms of the geographic location and spatial distribution of health services are important factors in healthcare utilization. In this descriptive paper we map the location of HIV related services and use exploratory spatial data analysis to visualize and examine the distribution of HIV service providers. In examining the location of HIV service providers we map the minimum distance to the nearest service provider. Our analyses also map and analyze five separate categories of HIV related services. These include: (1) Diagnostic and preventive services; (2) Health and social services for initial HIV diagnosis; (3) Emotional and social support; (4) Emergency services; and (5) Medical and end-of-life services. While our findings point to significant clustering of some types of HIV-related services (such as emergency and preventive services), other services are more evenly distributed across Toronto (this includes medical and end-of-life services). Our findings point to the need for policy makers and researchers to integrate mapping, GIS and spatial analytic techniques into their analyses of the neighborhoods and subsequently the populations in those neighborhoods that are underserved in terms of accessibility of some categories of HIV-related services. PMID- 15832888 TI - 'What I've noticed what they need is the stats': lay HIV counsellors' reports of working in a task-orientated health care system. AB - Counselling has been recognized as an important component of HIV and AIDS care, and an essential part of HIV testing. In South Africa, a commonly used model is for lay counsellors to be trained by non-governmental organizations and then to work alongside professionals in public health clinics. In studies of counselling in health care settings in the context of HIV, there has been a relative lack of attention to the organizational and systemic issues faced by counsellors and counselling programmes. Counsellors are involved in a dynamic interrelationship not only with their clients but also with the organizations in which they work. In this paper we report on counsellors' accounts of the impact of their unclear position on their work. Twenty-nine counsellors were interviewed using individual interviews and focus group discussions. The findings reveal a clash between an holistic counselling approach and a task-oriented health system. The results provide some indication of the need to consider workplace issues in planning and researching VCT. PMID- 15832889 TI - Brain injury. PMID- 15832890 TI - A preliminary investigation of road traffic accident rate after severe brain injury. AB - PRIMARY OBJECTIVE: to investigate the road traffic accident rate in patients who have resumed driving after severe brain injury. RESEARCH DESIGN: a retrospective study conducted by means of telephone interviews. METHODS AND PROCEDURES: The caregivers of 90 patients suffering from severe brain injury were included. All of the patients had sustained severe brain injury and prolonged coma, i.e. lasting for at least 48 hours. The caregivers were interviewed by means of a Questionnaire that investigated several aspects of driving competence after coma and the incidence of road traffic accidents. MAIN OUTCOMES AND RESULTS: All patient outcomes were evaluated by means of the Glasgow Outcome Scale (GOS). The 90 caregivers reported that 29 patients (32%) had resumed driving and that 11 of the 29 (38%) were subsequently involved in road traffic accidents. During the total duration of our patient population risk exposure, we found 11 cases in our study group, against the 4.7 expected cases calculated in the normal population. The relative risk of road traffic accidents in severe brain injury patients versus uninjured individuals was 2.3. CONCLUSIONS: Our preliminary data show that a subject who has suffered from severe brain injury and coma lasting for at least 48 hours has a statistically significant higher risk of being involved in a road traffic accident. PMID- 15832891 TI - Return to driving after traumatic brain injury: a British perspective. AB - PRIMARY OBJECTIVE: To identify the current legal situation and professional practice in assisting persons with traumatic brain injury (TBI) to return to safe driving after injury. METHODS AND PROCEDURES: A brief review of relevant literature, a description of the current statutory and quasi-statutory authorities regulating return to driving after TBI in the UK and a description of the nature and resolution of clinical and practical dilemmas facing professionals helping return to safe driving after TBI. Each of the 15 UK mobility centres was contacted and literature requested; in addition a representative of each centre responded to a structured telephone survey. MAIN OUTCOME AND RESULTS: The current situation in the UK is described, with a brief analysis of the strengths and weaknesses both of the current statutory situation, and also the practical situation (driving centres), with suggestions for improvements in practice. CONCLUSION: Although brain injury may cause serious limitations in driving ability, previous drivers are not routinely assessed or advised regarding return to driving after TBI. PMID- 15832892 TI - Predictive validity of driving-simulator assessments following traumatic brain injury: a preliminary study. AB - OBJECTIVE: To evaluate whether driving simulator and road test evaluations can predict long-term driving performance, we conducted a prospective study on 11 patients with moderate to severe traumatic brain injury. Sixteen healthy subjects were also tested to provide normative values on the simulator at baseline. METHOD: At their initial evaluation (time-1), subjects' driving skills were measured during a 30-minute simulator trial using an automated 12-measure Simulator Performance Index (SPI), while a trained observer also rated their performance using a Driving Performance Inventory (DPI). In addition, patients were evaluated on the road by a certified driving evaluator. Ten months later (time-2), family members observed patients driving for at least 3 hours over 4 weeks and rated their driving performance using the DPI. RESULTS: At time-1, patients were significantly impaired on automated SPI measures of driving skill, including: speed and steering control, accidents, and vigilance to a divided attention task. These simulator indices significantly predicted the following aspects of observed driving performance at time-2: handling of automobile controls, regulation of vehicle speed and direction, higher-order judgment and self-control, as well as a trend-level association with car accidents. Automated measures of simulator skill (SPI) were more sensitive and accurate than observational measures of simulator skill (DPI) in predicting actual driving performance. To our surprise, the road test results at time-1 showed no significant relation to driving performance at time-2. CONCLUSION: Simulator based assessment of patients with brain injuries can provide ecologically valid measures that, in some cases, may be more sensitive than a traditional road test as predictors of long-term driving performance in the community. PMID- 15832893 TI - Prevention of traffic accidents: the assessment of perceptual-motor alterations before obtaining a driving license. A longitudinal study of the first years of driving. AB - A longitudinal study was designed with two objectives: first, to provide a wide cognitive, personality and social description of new drivers before they started to drive cars. Second, to examine the relationship between cognitive and other characteristics drivers had before obtaining their driving license and the number and type of accidents they were involved in during the first years as drivers. RESEARCH DESIGN: The longitudinal study started in 1997 and ended in 2002. The first assessment was made up of 241 individuals at the time they enrolled on the driving course. The follow-up evaluation in the year 2002 was carried out on 144 components of the initial sample after five years driving. Age, gender and education level were matched to represent the population of Spain. METHOD AND PROCEDURES: Participants were assessed with the Bender Test for visual-motor ability, the B101 Test for practical intelligence, the B19 Test for visual-motor bi-manual coordination, and the TKK-1108 for speed anticipation. Personality was also evaluated with the Rorschach test and the PSY (Psychological Assessment Questionnaire). Five years later, a new examination of all those variables was made as well as a structured interview with each participant in order to collect data relating to significant life events during that time, driving habits, opinions in relation to certain traffic rules and information on accidents, incidents and/or sanctions. MAIN OUTCOMES AND RESULTS: Serious and/or minor accidents are concentrated on a few drivers. Accidentality is not related to gender or age, but educational level is related to serious accidents. The number of accidents (severe or minor ones) cannot be predicted if considered as a continuous variable, but it is possible if considered as a discrete variable. In this case two different cognitive profiles account for the number and type of accidents. CONCLUSION: The number and type of accidents during their first years of driving are related to the cognitive profiles of drivers assessed before they obtained their driving license. PMID- 15832894 TI - Role of premorbid factors in predicting safe return to driving after severe TBI. AB - PRIMARY OBJECTIVE: The present study explored the possibility of predicting post injury fitness to safe driving in patients with severe traumatic brain injury (TBI) (n = 66). METHODS AND PROCEDURE: Sixteen different measures, derived from four domains (demo/biographic, medico-functional, neuropsychological, and psychosocial) were used as predictor variables, whereas driving outcomes were assessed in terms of driving status (post-TBI drivers versus non-drivers) and driving safety (number of post-TBI car accidents and violations). MAIN OUTCOMES AND RESULTS: About 50% of the patients resumed driving after TBI. Compared to post-TBI non-drivers, post-injury drivers had shorter coma duration. With regard to driving safety, the final multiple regression model combined four predictors (years post-injury, accidents and violations before TBI, pre-TBI-risky personality-index, and pre-TBI-risky-driving-style-index) and explained 72.5% of variance in the outcome measure. CONCLUSIONS: Since the best three predictors of post-injury driving safety addressed patients' premorbid factors, the results suggest that in order to evaluate the actual possibility of safe driving after TBI, it would be advisable to consider carefully patients' pre-TBI histories. PMID- 15832895 TI - Driving with cognitive deficits: neurorehabilitation and legal measures are needed for driving again after severe traumatic brain injury. AB - PRIMARY OBJECTIVE: This article presents a retrospective study on a group of survivors of severe traumatic brain injury with the purpose of discerning whether post-traumatic cognitive deficits prevent them from safely resuming driving and to see if holistic neurorehabilitation improves the rate of patients fit for returning to driving. METHODS AND PROCEDURES: We studied 17 patients who had suffered severe traumatic brain injury (TBI) as measured by Glasgow Coma Scale scores. All subjects underwent a holistic, intensive and multidisciplinary neurorehabilitation program during a mean period of 10.53 months in the Centro de Rehabilitacion de Dano Cerebral (CRECER)--Center for Brain Injury Rehabilitation- in Seville, Spain. Patients were divided into two different groups: drivers (patients who drove despite strong and repeated recommendations from the Center to desist from doing so when they began the rehabilitation program) and non drivers (patients not driving at the time they began the rehabilitation program although they had a pre-injury driver's license). The FIM+FAM-Revised Scale [1] was administered both before commencing treatment and upon termination. RESULTS AND CONCLUSION: (1) Patients showing physical functionality above 80% returned to driving, regardless of their cognitive and/or emotional deficits, and against doctor recommendations. (2) Severe TBI survivors that have not been certified as fit to drive are at increased risk for driving incidents other than collisions and traffic accidents. This is illustrated by significant incidents involving some of the subjects in our study that were due to disorientation, confusion and confrontations with people or situations. (3) We found that neurorehabilitation is worthwhile; after integral and multidisciplinary neurorehabilitation more than 70% of survivors of severe TBI can return to driving with regular safety. (4) We also suggest that laws be introduced to keep not-clinically-apt patients from driving. PMID- 15832897 TI - Patient advocates: putting a face on spinal cord injury. PMID- 15832896 TI - Neuropsychological assessment of geriatric driving competence. AB - PRIMARY OBJECTIVE: To review of studies that focus on the assessment of driving competence among the elderly who are at increased risk of being involved in automobile crashes. The current status of neuropsychological testing as a predictor of driving safety in this population is critically evaluated. MAIN OUTCOMES AND RESULTS: Several domains of neuropsychological assessment have been shown to be related to safe driving in older age groups, including vision-based testing, attention-based testing, and testing of executive functions. Use of a driving simulator to investigate crash risk has been particularly effective. CONCLUSIONS: The argument is made that a combination of test approaches be used to develop an algorithm for efficient screening of elderly drivers on a regular basis and that use of a driving simulator to measure driving performance under challenge should be incorporated as part of this evaluation. PMID- 15832899 TI - Timing of surgery following spinal cord injury. AB - BACKGROUND: The optimal timing for surgical intervention after traumatic spinal injuries with spinal cord injury remains unclear. DESIGN: Literature review. FINDINGS: Multiple laboratory investigations (in animal models) and many clinical studies suggest better neurological outcomes with early surgical intervention. Conclusive evidence (well-designed randomized, controlled studies), however, is lacking, partly due to the logistics involved in executing such an investigation. Early surgery also appears to decrease the incidence of complications, reduces hospital stay, and helps reduce costs associated with acute management. CONCLUSION: Early surgical treatment is beneficial in terms of reducing complications, length of stay, and hospital costs. Further studies are needed to clearly demonstrate the impact of operative timing on neurological outcome. PMID- 15832900 TI - C-reactive protein, metabolic syndrome, and insulin resistance in individuals with spinal cord injury. AB - BACKGROUND/OBJECTIVE: High-sensitivity C-reactive protein (hsCRP) has been shown to be indicative of elevated risk of cardiovascular disease (CVD). We examined this blood marker in persons with spinal cord injury (SCI) in order to determine its potential for predicting elevated cardiac risk. METHODS: In a cross-sectional analysis, we examined the relationship between hsCRP and insulin resistance and metabolic syndrome among 93 individuals with SCI (50.2 +/- 13 years). Fasting lipids, insulin, glucose, plasma hsCRP, and anthropometric data were gathered from each subject. Comparisons were made with population reference values. RESULTS: Metabolic syndrome and insulin resistance were present in nearly one quarter of the SCI population (22.6%). Subjects with fasting insulin resistance had significantly higher mean hsCRP (4.29 +/- 3.25 mg/L) than those who were not insulin resistant (2.24 +/- 2.02) (P < 0.05). Moreover, hsCRP was significantly elevated in individuals who presented with high cardiovascular risk including severe dyslipidemia (> or = 4 abnormal lipid values) and Framingham Risk scores < or = 6 (P < 0.05 for both values). In addition, the homeostasis model assessment of insulin resistance was mildly correlated with hsCRP (r = 0.33). CONCLUSIONS: In individuals with SCI who are insulin resistant and/or display components of the metabolic syndrome, hsCRP is elevated suggesting a clinically important association with cardiovascular risk in this population. PMID- 15832901 TI - Upper limb strength in individuals with spinal cord injury who use manual wheelchairs. AB - INTRODUCTION: Manual wheelchair users have been found to be at risk for secondary upper extremity injuries. PURPOSE: The primary goal of this study was to compare shoulder strength and muscle imbalance of individuals with paraplegia to case wise matched unimpaired controls (UC). A secondary goal was to evaluate the impact of age and neurologic level of injury (NLI) on weight-normalized strength (WNS). METHODS: The SCI group (n = 28) and the UC group (n = 28) completed bilateral shoulder isokinetic strength testing in the sagittal, frontal, and horizontal plane at 60 degrees/second using the BioDex system. Strength ratios, an indicator of muscle imbalance, were also calculated. RESULTS: No significant difference was seen in shoulder strength or strength ratios between the SCI group and the UC group. However, NLI was significantly related to WNS on several planes in the SCI group. Therefore, we dichotomized the SCI group into equal groups based on an NLI. The Low-SCI group was significantly stronger than the High-SCI group in most planes (P < 0.05). The High-SCI group was significantly weaker than the UC in extension (P < 0.01) and a trend (P < 0.01) was seen in flexion, abduction, and external rotation. The Low-SCI group was significantly stronger in abduction than the UC. CONCLUSION: WNS at the shoulder correlated with NLI. It is likely that this is related to contributions of the trunk and abdominal muscles during testing, since proximal trunk strength aids in generating forces distally. This study and others of strength in individuals with paraplegia may overestimate shoulder strength. PMID- 15832902 TI - Implementation of clinical practice guidelines for prevention of thromboembolism in spinal cord injury. AB - BACKGROUND/OBJECTIVES: The purpose of this study was to determine whether publication of the "Prevention of Thromboembolism in Spinal Cord Injury" clinical practice guideline (CPG) changed patient management and whether adherence to CPG recommendations improved after a targeted implementation strategy. METHODS: Data were abstracted from medical records of 134 and 520 patients with acute and chronic spinal cord injury (SCI), respectively, from 6 Veterans Affairs medical centers over 3 time periods: prepublication (T1), preimplementation (T2), and postimplementation (T3) of the CPG. Targeted interventions were developed to address provider-perceived barriers to guideline adherence, based on findings from focus groups conducted at each site. The interventions incorporated two implementation strategies: standardized documentation templates/standing orders and social marketing/outreach visits. RESULTS: Use of the specified duration for pharmacologic prophylaxis increased from 60% to 65% to 75% of patients with acute SCI in T1, T2, and T3, respectively (P = 0.060 and 0.041 for T1 vs T2 and T2 vs T3, respectively). Rates of use for individual pharmacologic prophylaxis agents changed significantly over the course of the study, with use of low-molecular weight heparin increasing from 7% in T1 to 42% in T3. Physical assessments for thrombosis on hospitalization days 1 and 30 improved between T2 and T3. Use of prophylaxis in chronically injured patients with new risk factors for thromboembolism increased from 16% to 31% to 34% during T1, T2, and T3 (P = 0.001 and 0.87, respectively). CONCLUSIONS: The CPG publication had only a modest effect on practice. Use of structured implementation further increased the adherence to some CPG recommendations for thromboembolism prophylaxis. Similar implementation strategies should be considered for CPG recommendations with low adherence and high potential for morbidity and mortality. PMID- 15832903 TI - Preventive services and health behaviors among people with spinal cord injury. AB - BACKGROUND/OBJECTIVE: To document receipt of certain understudied preventive services and health behaviors in spinal cord injury (SCI) and to compare them to the general adult population. METHODS: Cross-sectional survey. PARTICIPANTS: One hundred ninety-nine community-living adults with SCI residing in New Jersey. MAIN OUTCOME MEASURES: Receipt of recommended general preventive health services, such as screening for colorectal and prostate cancer and dental examinations, and health-related behaviors, including injury prevention, obesity, and tobacco use, as assessed by survey items in the Behavioral Risk Factors Surveillance System. RESULTS: Deficiencies in provision of needed preventive services were evident but were similar to those noted in the general adult population. Among participants 50 years of age or older, 47% had not had either a stool test for occult blood in the last year or a sigmoidoscopy/colonoscopy in the last 5 years. While 43.9% of participants were overweight or obese and 46.3% of these were trying to lose weight, only 1 3.8% had been counseled to lose weight. Twenty-two percent smoked on at least some days. Safety problems were evident: 15.5% did not use seatbelts consistently; 44% had not discussed a fire escape plan at home; and smoke detectors were often not checked adequately. Twelve and a half percent of participants reported having injuries during the last year, and fully 75% of these were related to SCI. Emergency room use and hospitalization were frequent. Participants had a median of 3 to 5 current doctors. CONCLUSIONS: Persons with SCI need the general screening and safety services recommended for all persons. Enhanced efforts will be needed to meet national goals in Healthy People 2010 in SCI. PMID- 15832904 TI - Bilateral oculosympathetic paresis associated with loss of nocturnal melatonin secretion in patients with spinal cord injury. AB - BACKGROUND: Lesions along the sympathetic pathway to the eye produce oculosympathetic paresis (OSP, Horner's syndrome). The oculosympathetic pathway descends from the hypothalamus through the cervical spinal cord and ascends to the superior cervical ganglion (SCG), which innervates sympathetic targets in the ipsilateral face and eye. This pathway appears to closely co-localize with a similar retino-pineal neural pathway from the hypothalamus through the cervical spinal cord and SCG to the pineal gland. As such, lesions along this shared pathway, such as occur in neurologically complete injury to the cervical spinal cord (tetraplegia), would be predicted to result in simultaneous OSP and loss of pineal melatonin production. Loss of melatonin production may contribute to the pervasive sleep disruption observed in patients with tetraplegia. METHODS: We assessed the presence of OSP by photographic documentation of ptosis and pupillary dilation response to cocaine eye drops in 5 individuals with neurologically complete damage to their upper thoracic or lower cervical spinal cord. We correlated these results with an analysis of the pattern of melatonin production in these same individuals. RESULTS: Bilateral OSP was present in individuals with cervical spinal cord injury; each also lacked significant production of melatonin. No evidence of OSP was observed in the 2 individuals with thoracic spinal cord injury below the level of the oculosympathetic pathway. Both had normal circadian rhythms of melatonin production, with timing and amplitude of the rhythm within normal parameters. CONCLUSION: The presence of bilateral oculosympathetic paresis can be predictive of the complete loss of the nocturnal production of melatonin. PMID- 15832905 TI - Diabetes mellitus and preventable spinal cord injury. AB - PURPOSE: A high prevalence of diabetes mellitus (DM) among spinal cord injury (SCI) populations has been noted. A review and follow-up of acute admissions to a SCI service was conducted to explain this high prevalence. METHODS: Records of 166 patients admitted for rehabilitation within 108 days of paralysis were reviewed. Cases of DM were identified by records of treatment or fasting blood glucose levels > 140 mg% or hemoglobin A1c < 7 g%. The onset of DM relative to SCI, the age at injury, and the cause of injury were recorded. New cases of DM and all deaths within a 5-year follow-up were determined for those patients 40 years of age or older at paralysis. RESULTS: The prevalence of DM was 0 of 79 younger patients (aged 16 to 39 years) but 18 of 87 older patients (aged 40 to 86 years), or 21%. All patients with DM had been diagnosed before SCI (16 patients) or during the initial hospitalization (2 patients). The cause of injury was falling in 11 of 18 (61%) DM patients and 31 of 69 (45%) nondiabetic older patients (P = 0.29). Falling was a less common cause of injury in the younger group (13 of 78 [17%], P < 0.001). The cause of paralysis was medical (ie, vascular, infectious) in 5 patients in the DM group (28%) and 5 patients in the non-DM group (6%) (P = 0.03). The 5-year mortality for patients with DM was 7 of 17 (one lost to follow-up) (41%) and 10 of 64 (16%; 5 lost to follow-up) for the nondiabetic older patients (P = 0.04). The 5-year prevalence of DM in the survivors (4 patients developed DM, 1 patient did not survive) was 23%, similar to the initial prevalence. CONCLUSION: DM is a risk factor for SCI. Attention to this risk could prevent some SCIs. PMID- 15832906 TI - Study of the rectal electric activity of the uninhibited rectal detrusor (overactive rectum): a new concept of pathogenesis. AB - BACKGROUND/OBJECTIVE: One of the causes of fecal incontinence is uninhibited rectal detrusor syndrome (URDS). Patients with this condition either perceived the first rectal sensation after the onset of involuntary rectal contraction or not at all. We investigated the hypothesis that the abnormal rectal contractility in URDS may be caused by deranged rectal electric activity. METHODS: Twenty-five patients with URD (14 women and 11 men; age, 44.7 +/- 10.3 years) and 10 healthy volunteers (6 women and 4 men; age, 42.8 +/- 8.7 years) were studied. URDS was diagnosed by rectometry and provocative test. A transcutaneous EMG was performed with one electrode placed lateral to each sacroiliac joint and the third one midway between the greater trochanter and the ischial tuberosity. Two 20-minute recording sessions were performed for each subject. RESULTS: Slow waves (SWs) with regular rhythm and similar parameters (frequency, amplitude, conduction velocity) from the 3 electrodes were recorded from the healthy volunteers. They showed a significant increase in the parameters on saline filling of the rectum. The SWs of patients with URDS exhibited a "dysrhythmic" pattern with irregular parameters, which were different in the 3 electrodes and inconsistent during recording. They showed areas of tachyrhythmia, bradyrhythmia, and arrhythmia. On provoking rectal overactivity, the SWs showed an increased dysrhythmic activity. CONCLUSIONS: The patients with URD exhibited a "dysrhythmic" electric pattern with areas of variable electric activity. The tachyrhythmic areas seem to initiate the urgency and fecal incontinence of URDS. It is suggested that a disordered rectosigmoid pacemaker causes the dysrhythmic waves. PMID- 15832907 TI - Anterior spinal artery syndrome in two children with genetic thrombotic disorders. AB - BACKGROUND: Spinal cord infarction is a well-described, but rare, etiology of myelopathy, especially in children. The most common syndrome, anterior spinal artery syndrome (ASAS), is caused by interruption of blood flow to the anterior spinal artery, producing ischemia in the anterior two-thirds of the cord, with resulting neurologic deficits. Causes of ASAS include aortic disease, thoracolumbar surgery, sepsis, hypotension, and thromboembolic disorders. METHODS: Case reports of 2 patients. RESULTS: Two children developed spinal cord infarctions consistent with ASAS, mostly likely caused by previously undiagnosed thrombotic disorders. A child with prothrombin variant experienced acute bilateral lower limb weakness without any preceding event. Magnetic resonance imaging (MRI) revealed increased T2 signal in the anterior cord from midthoracic level to the conus medullaris. A child with protein S deficiency developed lower limb weakness 1 day after a posterior thoracolumbar fusion for idiopathic scoliosis. Computed tomography (CT) myelogram revealed no spinal cord compression. The prothrombin variant mutation is associated with a 2-fold risk of thrombotic events. Individuals with protein S deficiency have an 8-fold increased risk of thrombosis. CONCLUSION: As knowledge of the coagulation pathways grows, it is likely that more patients with spinal cord infarctions will be diagnosed with genetic thrombotic disorders as the etiology of their injury. We review these two disorders, prothrombin variant and protein S deficiency, and the considerations for long-term anticoagulation. PMID- 15832908 TI - Concept for a laser guide beacon Shack-Hartmann wave-front sensor with dynamically steered subapertures. AB - We describe an innovative implementation of the Shack-Hartmann wave-front sensor that is designed to correct the perspective elongation of a laser guide beacon in adaptive optics. Subapertures are defined by the segments of a deformable mirror rather than by a conventional lenslet array. A bias tilt on each segment separates the beacon images on the sensor's detector. One removes the perspective elongation by dynamically driving each segment with a predetermined open-loop signal that would, in the absence of atmospheric wave-front aberration, keep the corresponding beacon image centered on the subaperture's optical axis. PMID- 15832909 TI - Controllable double-well magneto-optic atom trap with a circular current-carrying wire. AB - We propose a controllable double-well magneto-optic trap (MOT) for neutral atoms that uses a circular current-carrying wire and a biased magnetic field. One can cause the proposed double-well trap to evolve continually into a single-well trap by reducing the current in the wire or increasing the bias field. We estimate that in a weak intensity approximation a cold atomic sample with a temperature of approximately 300 K and a number of approximately 10(6) atoms can be stored in each MOT. One can control the number of trapped atoms by changing the current in the wire. PMID- 15832910 TI - All-optical switching and routing based on an electromagnetically induced absorption grating. AB - An electromagnetically induced absorption grating is formed in a three-level atomic vapor under the condition of electromagnetically induced transparency in which the strong coupling beam is replaced by a standing wave. The transmission and reflection behaviors of the weak probe beam are greatly modified at certain frequencies near the two-photon resonance. An all-optical two-port signal router all-optical switch is demonstrated. PMID- 15832911 TI - Photonic phase shifters based on a vector-sum technique with polarization maintaining fibers. AB - We propose and demonstrate a photonic phase shifter based a on vector-sum technique that uses polarization-maintaining fibers (PMFs). We achieved a continuous and full phase shift up to 2pi at 30.48 GHz by controlling the polarization state of the light injected into two pieces of PMF of different lengths and applying two different modulator bias voltages. PMID- 15832912 TI - Brillouin spectral deconvolution method for centimeter spatial resolution and high-accuracy strain measurement in Brillouin sensors. AB - Combining a dc and a short pulse (approximately 1 ns) as the probe beam in the pump-probe configuration of Brillouin-based distributed sensors allows us to represent the Brillouin spectrum as a top Lorentzian-like portion and a bottom Gaussian-like portion. Because of the interaction of these two parts, the Lorentzian-like portion carries spatial information that can be extracted within centimeter spatial resolution. Using this information, we develop a spectrum deconvolution method, which considers the location correlation of the strain distribution, to find the number of Brillouin peaks and their frequencies in the top Lorentzian-like portion and hence achieve accurate strain information. An optimum level of dc to pulse power for the best signal and position detection capability is discussed. PMID- 15832913 TI - Photothermal effects in passive fiber Bragg grating resonators. AB - Photothermal effects in passive Fabry-Perot resonators are caused by the conversion of circulating optical energy into heat as a result of absorption. This results in thermal change in the resonator's optical path length, the round trip phase, and hence the resonance condition. We describe a simplified dynamic numerical model for photothermal effects in passive fiber Bragg grating resonators and present results of their experimental observation. PMID- 15832914 TI - Sapphire-fiber-based white-light interferometric sensor for high-temperature measurements. AB - We present a fiber-optic temperature sensor with a single-crystal sapphire fiber as the light guide and a sapphire wafer as the sensing element. Temperature is determined by measurement of the thermal dependence of the wafer's optical thickness by use of white-light interferometry. We applied digital signal processing techniques to analyze the sensor's spectrum. A prototype sensor was tested to 1600 degrees C and demonstrated excellent reproducibility. An accuracy of +/- 0.2% full scale was obtained. The sensor is simple, small, and flexible, with potential advantages of batch fabrication and easy calibration. PMID- 15832915 TI - High-power picosecond fiber amplifier based on nonlinear spectral compression. AB - We report on the rare-earth-doped fiber-based generation of nearly transform limited 10-ps pulses based on self-phase-modulation-induced spectral compression. An ytterbium-doped low nonlinearity photonic crystal fiber is used as a gain medium. An average power of as much as 97 W at a repetition rate of 47 MHz, corresponding to a peak power as high as 200 kW, was obtained. Furthermore, efficient second-harmonic generation by application of this high-power laser source is discussed. PMID- 15832916 TI - Selective mode excitation in hollow-core photonic crystal fiber. AB - Modes are selectively excited by launching light through the cladding from the side into a hollow-core photonic crystal fiber. Measuring the total output power at the end of the fiber as a function of the angle of incidence of the exciting laser beam provides a powerful diagnostic for characterizing the cladding bandgap. Furthermore, various types of modes on either side of the bandgap are excited individually, and their near-field images are obtained. PMID- 15832917 TI - Deposition of overlays by electrostatic self-assembly in long-period fiber gratings. AB - It was proved that the deposition of an overlay material onto a long-period fiber grating causes important shifts in the wavelengths of the typical attenuation bands that are caused by coupling between cladding and core modes [Opt. Lett. 27, 682 (2002)]. A theoretical model for analyzing a multilayer cylindrical waveguide is presented that permits the phenomenon to be understood and predicted. An overlay of higher refractive index than the cladding starts to guide a mode if a certain thickness value is exceeded. This causes large shifts in the resonance wavelength induced by the grating. One important application of this phenomenon to sensors is enhancement of the sensitivity of a long-period fiber grating to ambient conditions. Theoretical results are corroborated with experimental ones obtained by electrostatic self-assembly. PMID- 15832918 TI - Low-loss waveguides written with a femtosecond laser for flexible interconnection in a planar light-wave circuit. AB - We describe a low-loss single-mode waveguide in planar light-wave circuit (PLC) glass doped with boron and phosphorus, which is more difficult to write than pure silica glass. The written waveguide has a rectangular core, a symmetric near field pattern, and a propagation loss of 0.35 dB/cm. The loss that originates from the mode-field mismatch between the mode-field diameters of the written and the PLC waveguides is less than 0.1 dB/point. In addition, we successfully connected PLC waveguides with a 500-microm-long waveguide written with a laser. The laser-written waveguide can flexibly connect PLC waveguides with a low coupling loss. PMID- 15832919 TI - 520-W continuous-wave diode corner-pumped composite Yb:YAG slab laser. AB - We present a pumping scheme for a quasi-three-level solid-state laser. The scheme uses a slab laser configuration with the pump light incident from the slab corners. A diode-corner-pumped composite Yb:YAG-YAG slab laser operating in high power cw mode was designed to prove the scheme. As much as 520 W of output power was obtained from a single 1-mm-thick composite slab with 0.5-at. %-doped Yb:YAG. The slope efficiency and the optical-to-optical efficiency with respect to the pump power were 32% and 25%, respectively. This result shows the validity of the corner-pumping concept and its feasibility in the development of high-power solid state lasers. PMID- 15832920 TI - Laser activity at 1.18, 1.07, and 0.97 microm in the low-phonon-energy hosts KPb2Br5 and RbPb2Br5 doped with Nd3+. AB - For the first time to the authors' knowledge, laser activity has been achieved in low-phonon-energy, moisture-resistant bromide host crystals, neodymium-doped potassium lead bromide (Nd3+:KPb2Br5) and rubidium lead bromide (Nd3+:RbPb2Br5; RPB). Laser activity at 1.07 microm was observed for both crystalline materials. Laser operation at the new wavelengths 1.18 and 0.97 microm that resulted from the 4F5/2 + 2H9/2 - 4IJ transitions (J=13/2 and J=11/2) in Nd:RPB was achieved in a solid-state laser material. Rare-earth-doped MPb2Br5 (M=K, Rb) is a promising candidate for long-wavelength infrared applications because of its low phonon frequencies and other favorable features. In principle, Nd3+:MPb2Br5 has high potential for laser operation at new wavelengths as well as for the achievement of short-wavelength lasing as a result of upconversion. PMID- 15832921 TI - Experimental studies of spontaneous emission from dopants in an absorbing dielectric. AB - We report the first measurements, to our knowledge, of the modification of spontaneous emission rates of Eu3+ ions in the visible region owing to an absorbing medium. Precise levels of the absorption coefficient are introduced by codoping with different amounts of Nd3+. We use a binary glass system PbO-B2O3 as the host, the compositional variation of which leads to a change in the real part of the refractive index. Measured lifetimes are found to follow the real cavity model, and the data are analyzed by the model proposed by Scheel et al. [Phys. Rev. A 60, 4094 (1999)]. We give estimates of the parameter that is related to the radius of the cavity around Eu3+. PMID- 15832922 TI - Solid-state carrier-envelope phase stabilization via quantum interference control of injected photocurrents. AB - We demonstrate carrier-envelope phase stabilization of a mode-locked Ti:sapphire laser by use of quantum interference control of injected photocurrents in a semiconductor. No harmonic generation is required for this stabilization technique. Instead, interference between coexisting single- and two-photon absorption pathways in the semiconductor provides a phase comparison between different spectral components. The phase comparison, and the detection of the photocurrent that it produces, both occur within a single low-temperature-grown gallium arsenide sample. The carrier-envelope offset beat note fidelity is 30 dB in a 10-kHz resolution bandwidth. The out-of-loop phase-noise level is essentially identical to the best previous measurements with the standard self referencing technique. PMID- 15832923 TI - Deciphering output coupling mechanisms in spiral microcavities with femtosecond light bullets. AB - A pump-probe two-photon-excited fluorescence technique deciphers in space and time the propagation of ballistic wave packets sustained by whispering-gallery modes (WGMs) in a spiral-shaped microcavity. Diffraction on the spiral discontinuity does not prevent the WGMs from closing. The resultant average Q of the resonator is 3 x 10(4) +/- 50%. Experimental results are compared with numerical simulations, providing evidence of a new contribution to output coupling: Part of the WGM evanescent wave is reflected at the spiral notch and leads to a propagating wave at an angle that matches the previously observed laser emission direction in 4-dicyanomethylene-2-methyl-6-(p-dimethylaminostyryl) 4H-pyran-doped poly(methyl methacrylate) and InGaN spiral lasers. PMID- 15832924 TI - Depth-resolved low-coherence enhanced backscattering. AB - The phenomenon of enhanced backscattering (also known as coherent backscattering), an object of substantial scientific interest, has awaited application to tissue optics for the past two decades. Here we demonstrate, for the first time to our knowledge, depth-resolved spectroscopic elastic light scattering measurements in tissue by use of low-coherence enhanced backscattering (LEBS). We achieve the depth resolution by exploiting the nature of the LEBS peak that contains information about a wide range of tissue depths. We further demonstrate that depth-resolved LEBS spectroscopy has the potential to identify the origin of precancerous transformations in the colon at an early, previously undetectable stage. PMID- 15832925 TI - Fusion of conventional ultrasound imaging and acousto-optic sensing by use of a standard pulsed-ultrasound scanner. AB - Acousto-optic sensing (AOS) is a dual-wave sensing technique based on the ultrasound modulation of diffuse light in a turbid medium. We experimentally demonstrate the feasibility of combining AOS and conventional ultrasound imaging by use of a commercially available pulsed-ultrasound scanner coupled with a photorefractive crystal-based optical interferometry system. Optically absorbing targets embedded in highly diffusive phantoms (mus'= 10 cm(-1)) are imaged through a thickness of 27 mm with millimeter resolution. The acousto-optic images are intrinsically coregistered with the ultrasound images. PMID- 15832926 TI - Imaging magnetically labeled cells with magnetomotive optical coherence tomography. AB - We introduce a novel contrast mechanism for optical coherence tomography (OCT) whereby the optical scattering of magnetically labeled cells is modified by means of an externally applied magnetic field. This modification is made through the addition of a small electromagnet to the imaging arm of a conventional OCT interferometer. We measure the magnetomotive OCT signal by differencing pairs of axial scans (A-scans) acquired with the magnetic field on and off. Magnetomotive contrast is demonstrated in bulk three-dimensional cell scaffolds containing macrophages labeled with microparticles of iron oxide, demonstrating magnetic specific contrast over a dynamic range of 30 dB. PMID- 15832927 TI - Dynamic instability of self-induced bidirectional waveguides in photorefractive media. AB - We report on the experimental observation of a dynamic instability in the interaction of counterpropagating self-trapped beams in a photorefractive strontium barium niobate crystal. While the interaction of copropagating spatial optical solitons exhibits only transient dynamics, resulting in a final steady state, the counterpropagating geometry supports a dynamic instability mediated by intrinsic feedback. Experimental observations are compared with and found to be in qualitative agreement with numerical simulations. PMID- 15832928 TI - Variation of the carrier-envelope phase of few-cycle laser pulses owing to the Gouy phase: a solid-state-based measurement. AB - The carrier-envelope phase of a laser pulse has recently become an important quantity in extreme nonlinear optics. Because of the topological Gouy phase, it changes while the pulse propagates through the focus of a lens. This variation is measured by a simple solid-state-based approach. The experimental results are analyzed by comparison with simple analytical model calculations. PMID- 15832929 TI - Route to broadband blue-light generation in microstructured fibers. AB - We explore theoretically the possibility of generating broadband blue light by copropagating a short soliton pump pulse and a broader signal pulse in a microstructured fiber with a zero-dispersion wavelength located between the center wavelength of the pump and the signal pulses. We show that the unique properties of microstructured fibers should allow for broadening of the signal pulse's spectrum by as much as a factor of 50 through the conjugate action of cross-phase modulation and a soliton self-frequency shift. The physical mechanism that leads to this large spectral broadening is analyzed by use of an extended nonlinear Schrodinger equation. PMID- 15832930 TI - Widely tunable optical parametric generation in a photonic crystal fiber. AB - We report on the observation of widely tunable optical parametric generation in a photonic crystal fiber. The frequency shift of the generated sidebands that arise from modulational instability is strongly dependent on the detuning of the pump from the fiber's zero-dispersion wavelength. We are able to demonstrate experimentally more than 450 nm of sideband tunability as we tune the pump wavelength over 10 nm. Excellent agreement has been found between the experimentally measured and theoretically predicted shifts. PMID- 15832931 TI - High-contrast interference pattern of amplified spontaneous emission from dyes under transient grating excitation. AB - We demonstrate a high-contrast interference fringe pattern formed by amplified spontaneous emission (ASE) in a noncollinear two-beam self-diffraction setup by use of dyes. In the presence of a transient grating a stable interference pattern is observed in ASE induced by one- and two-photon absorption. The fringe visibility varies from 0.2 to 0.65 with a temporal delay in the two beams. The variation of the angle between the two exciting beams at the temporal overlap shows an increase in the fringe visibility with an increase in the angle. PMID- 15832932 TI - High-order harmonic generation from boron plasma in the extreme-ultraviolet range. AB - We demonstrate the generation of up to the 63rd harmonic (lambda = 12.6 nm) of a Ti:sapphire laser pulse (150 fs, 10 mJ), using a prepulse- (210-ps,24-mJ) produced boron plasma as the nonlinear medium. The influence of various parameters on the harmonic conversion efficiency was analyzed. The steep decrease in intensity for low-order harmonics (up to 19th order) was followed by a plateau. Typical conversion efficiencies were evaluated to be 10(-4) (for a 3rd harmonic) to 10(-7) (within the plateau region). Harmonic generation appeared to be efficient for the plasma that comprised neutral atoms and singly ionized boron. PMID- 15832933 TI - Compensation for pixel-mismatch errors in shift-multiplexed holographic data storage. AB - To read out correct data from shift-multiplexed holographic data storage, we propose a novel method of pixel matching by use of a tiny plane-parallel refraction plate placed in the path of converging readout beams. Tilting the refraction plate produces a shift in the lateral position of the readout beam incident upon the holographic storage disk, leading to compensation for pixel mismatch errors by an improvement in bit-error rate of 3 orders of magnitude. PMID- 15832934 TI - Sm(DBM)3Phen-doped poly(methyl methacrylate) for three-dimensional multilayered optical memory. AB - We report on the formation of submicrometer voids within Sm(DBM)3Phen-doped poly(methyl methacrylate) (PMMA) under multiphoton absorption excited by an infrared laser beam. An ultrashort-pulsed laser beam with a pulse width of 200 fs at a wavelength of 800 nm is focused into doped PMMA. The large changes in refractive index and the fluorescence associated with a void allow conventional optical microscopy and reflection-type confocal microscopy to be used as detection methods. Voids can be arranged in a three-dimensional multilayered structure for high-density optical data storage. PMID- 15832935 TI - Generation of flat power-envelope terahertz-wide modulation sidebands from a continuous-wave laser based on an external electro-optic phase modulator. AB - Flat power-envelope terahertz-wide modulation sidebands are generated by only electro-optic phase modulation of continuous-wave laser light. Generation and power equalization of widespread sidebands are realized simultaneously by spatial distribution of the modulation index within a laser beam cross section by use of simple domain-engineering processes in LiTaO3 electro-optic crystal. Generation of 46 sidebands spaced by 16.25 GHz within a -3-dB bandwidth (over a 1-THz span for a +/- 3-dB bandwidth) is demonstrated. PMID- 15832936 TI - Suppression of radiative losses of surface polaritons on nanostructured thin metal films. AB - The strong electromagnetic coupling between surface plasmon polariton modes on opposite interfaces of a finite thickness periodically nanostructured metal film has been studied. Surface polariton dispersion and associated electromagnetic field distributions have been analyzed. It was shown that at a frequency that corresponds to the crossing of film Bloch modes of different symmetries, the radiative losses of surface polaritons that are related to the polaritons' coupling to light during propagation on the structured surface are suppressed. PMID- 15832937 TI - Laser-induced forces between carbon nanotubes. AB - Carbon nanotubes are the focus of intense research interest because of their unique properties and applications potential. We present a study based on quantum electrodynamics concerning the optical force between a pair of nanotubes under laser irradiance. To identify separate effects associated with the pair orientation and laser beam geometry, two different systems are analyzed. For each, an analytical expression for the laser-induced optical force is determined, and the corresponding magnitude is estimated. PMID- 15832938 TI - Dynamic localization and Bloch oscillations in the spectrum of a frequency mode locked laser. AB - It is shown that a frequency mode-locked laser with a sinusoidal sweep of modulation frequency around a mode-locking condition represents an ideal optical system for observing in the spectral domain the phenomena of dynamic localization and Bloch oscillations of electrons in an ideal solid placed in an external ac electric field. PMID- 15832939 TI - Phase-noise properties of an ytterbium-doped fiber amplifier for the Laser Interferometer Space Antenna. AB - Phase noise is a critical issue for the sensitivity of the interferometric gravitational-wave detector Laser Interferometer Space Antenna that requires laser sources with 1 W of output power. A low-power seed laser with a subsequent fiber amplifier is in discussion, and we report on spectrally resolved phase noise measurements of an ytterbium-doped fiber amplifier emitting 1 W of output power from 10 mW of seed power. Phase-noise measurements for Fourier frequencies from 10(-4) Hz to 100 kHz show 60 rad/Hz0.5 at 10(-4) Hz and 0.2 mrad/Hz0.5 at 1 kHz. This measured excess phase noise is orders of magnitude below the free running phase noise of nonplanar ring oscillators. PMID- 15832940 TI - Theory of light propagation incorporating scattering and absorption in turbid media. AB - A general theoretical approach to the description of light propagating through turbid media is proposed. The theory is a modification of the two-flux model of Kubelka-Munk (KM), extending its applicability to media systems containing an absorptive component. The modified KM model takes into account the influence of internal scattering on the total path length and accommodates a wide range of absorption influences. Experimental results obtained for dyed-paper systems illuminated by diffuse light are demonstrated to be qualitatively and quantitatively reproduced by the theory. PMID- 15832941 TI - Adaptive correction of human-eye aberrations in a subjective feedback loop. AB - An adaptive optical system with a subjective feedback loop is used to improve the visual acuity and to determine the aberrations of the human eye. Corrections of as many as 12 low-order aberration modes were made, based on the perceived sharpness of the test object observed through the adaptive optical system. The acuity of vision was improved by adjustment of the weights of the orthogonal modes produced by a deformable mirror. Objective measurements of the correcting aspherical figures, obtained in independent subjective correction cycles for one person, demonstrated good repeatability. Participants in the study with strong ocular aberrations reported moderate to significant improvement of their visual acuity, estimated with the U.S. Air Force 1951 acuity chart. PMID- 15832942 TI - Expression of mdr49 and mdr65 multidrug resistance genes in larval tissues of Drosophila melanogaster under normal and stress conditions. AB - In situ expression of 2 multidrug resistance genes, mdr49 and mdr65, of Drosophila melanogaster was examined in wild-type third instar larval tissues under physiological conditions and after heat shock or colchicine feeding. Expression of these 2 genes was also examined in tumorous tissues of lethal (2) giant larvae I(2)gl4 mutant larvae. These 2 mdr genes show similar constitutive expression in different larval tissues under physiological conditions. However, they are induced differentially by endogenous (tumorous growth) and exogenous stresses (colchcine feeding or heat shock): whereas heat shock and colchicine feeding induce mdr49, tumorous condition is accompanied by enhanced expression of mdr49 and mdr65 genes. PMID- 15832943 TI - Use of surface-enhanced laser desorption ionization-time-of-flight to identify heat shock protein 70 isoforms in closely related species of the virilis group of Drosophila. AB - The 70-kDa heat shock protein (Hsp) family in all Drosophila species includes 2 environmentally inducible family members, Hsp70 and Hsp68. Two-dimensional gel electrophoresis revealed an unusual pattern of heat shock-inducible proteins in the species of the virilis group. Trypsin fingerprinting and microsequencing of tryptic peptides using ProteinChip Array technology identified the major isoelectric variants of Hsp70 family, including Hsp68 isoforms that differ in both molecular mass and isoelectric point from those in Drosophila melanogaster. The peculiar electrophoretic mobility is consistent with the deduced amino acid sequence of corresponding hsp genes from the species of the virilis group. PMID- 15832944 TI - Effects of hypoxia on stress proteins in the piglet heart at birth. AB - Hypoxia at birth represents a very stressful event that can result in severe lifelong consequences in different tissues, including those of the heart. Heat shock and other associated stress proteins are involved in cellular protection, but their roles are not clearly defined at the time of birth. Newborn piglets were subjected to 5% oxygen and 95% nitrogen for either 1 or 4 hours. They were allowed to recover over periods of 1 to 68 hours. The relative levels of alphaB crystallin, HspB8, Hsp20, Hsp27, Hsp60, and Hsp70 as well as nitric oxide synthases (NOS) (endothelial NOS, inducible NOS, neuronal NOS) were examined by Western blot analysis. Surprisingly, alphaB-crystallin expression was drastically increased in animals submitted to hypoxia. The hypoxia-associated factor HIFlalpha was also strongly and rapidly overexpressed. Heme oxygenase 1 was also increased. To a lesser extent, neuronal NOS was also increased in the left ventricle of animals submitted to hypoxia. This work clearly shows that the Hsp chaperone alphaB-crystallin is strongly overexpressed in the left ventricle of animals submitted to hypoxia. This observation dissociates the response to low oxygenation of alphaB-crystallin and other stress-associated proteins including Hsp27, and it indicates that heme oxygenase is not alone among HSPs in its oxygen related gene expression. PMID- 15832945 TI - Factors governing the substrate recognition by GroEL chaperone: a sequence correlation approach. AB - The chaperonin GroEL binds to a large number of polypeptides, prevents their self association, and mediates appropriate folding in a GroES and adenosine triphosphate-dependent manner. But how the GroEL molecule actually recognizes the polypeptide and what are the exact GroEL recognition sites in the substrates are still poorly understood. We have examined more than 50 in vivo substrates as well as well-characterized in vitro substrates, for their binding characteristics with GroEL. While addressing the issue, we have been driven by the basic concept that GroES, being the cochaperonin of GroEL, is the best-suited substrate for GroEL, as well as by the fact that polypeptide substrate and GroES occupy the same binding sites on the GroEL apical domain. GroES interacts with GroEL through selective hydrophobic residues present on its mobile loop region, and we have considered the group of residues on the GroES mobile loop as the key element in choosing a substrate for GroEL. Considering the hydrophobic region on the GroES mobile loop as the standard, we have attempted to identify the homologous region on the peptide sequences in the proteins of our interest. Polypeptides have been judged as potential GroEL substrates on the basis of the presence of the GroES mobile loop-like hydrophobic segments in their amino acid sequences. We have observed 1 or more GroES mobile loop-like hydrophobic patches in the peptide sequence of some of the proteins of our interest, and the hydropathy index of most of these patches also seems to be approximately close to that of the standard. It has been proposed that the presence of hydrophobic patches having substantial degree of hydropathy index as compared with the standard segment is a necessary condition for a peptide sequence to be recognized by GroEL molecules. We also observed that the overall hydrophobicity is also close to 30% in these substrates, although this is not the sufficient criterion for a polypeptide to be assigned as a substrate for GroEL. We found that the binding of aconitase, alpha lactalbumin, and murine dihydrofolate reductase to GroEL falls in line with our present model and have also predicted the exact regions of their binding to GroEL. On the basis of our GroEL substrate prediction, we have presented a model for the binding of apo form of some proteins to GroEL and the eventual formation of the holo form. Our observation also reveals that in most of the cases, the GroES mobile loop-like hydrophobic patch is present in the unstructured region of the protein molecule, specifically in the loop or beta-sheeted region. The outcome of our study would be an essential feature in identifying a potential substrate for GroEL on the basis of the presence of 1 or more GroES mobile loop like hydrophobic segments in the amino acid sequence of those polypeptides and their location in three-dimensional space. PMID- 15832946 TI - Clonogenicity of human leukemic cells protected from cell-lethal agents by heat shock protein 70. AB - Pretreatment of human leukemia THP-1 cells with heat shock protein Hsp70 (Hsp70) protected them from the cell-lethal effects of the topoisomerase II inhibitor, lucanthone and from ionizing radiation. Cell viability was scored in clonogenic assays of single cells grown in liquid medium containing 0.5% methyl cellulose. Colonies were observed and rapidly scored after staining with the tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide. The frequency of abasic sites in the deoxyribonucleic acid (DNA) of THP-1 cells was reduced when these cells were treated with Hsp70. Hsp70 is presumed to have protected the cells by promoting repair of cell DNA, in agreement with previous studies that showed that Hsp70 enhanced base excision repair by purified enzymes. The shoulders of radiation dose-response curves were enhanced by pretreatment of cells with Hsp70 and, importantly, were reduced when cells were transfected with ribonucleic acid designed to silence Hsp70. Hsp70 influenced repair of sublethal damage after radiation. PMID- 15832948 TI - CD95-mediated alteration in Hsp70 levels is dependent on protein stabilization. AB - Engagement of death receptors induces caspase activation and apoptosis. A recent study reported altered protein expression, including increased Hsp70 levels during CD95-mediated apoptosis. Here, we examined the mechanism underlying increased Hsp70 levels in cells challenged with a monoclonal antibody directed against the CD95 receptor. Levels of Hsp70 were found to increase in a dose dependent manner, occurring independently of either heat shock factor 1 activation or the accumulation of Hsp70 messenger ribonucleic acid (mRNA), suggesting the involvement of posttranslational modifications. Inhibition of translation and de novo protein synthesis by cycloheximide resulted in Hsp70 protein levels diminishing over time in control cells, whereas its level remained constant during CD95 signaling. In addition, death receptor activation through exposure of cells to tumor necrosis factor-related apoptosis-inducing ligand did not alter Hsp70 levels. These findings demonstrate that receptor-specific signaling through the CD95 increases the stability of Hsp70 protein, rather than mRNA, when compared with control cells. The results describe a novel mechanism of heat shock protein accumulation, where increased protein stability and reduced turnover, is the mechanism by which Hsp70 accumulates in cells during CD95 mediated apoptosis. PMID- 15832947 TI - Expression of heat shock proteins and heat shock protein messenger ribonucleic acid in human prostate carcinoma in vitro and in tumors in vivo. AB - Heat shock proteins (HSPs) are thought to play a role in the development of cancer and to modulate tumor response to cytotoxic therapy. In this study, we have examined the expression of hsf and HSP genes in normal human prostate epithelial cells and a range of prostate carcinoma cell lines derived from human tumors. We have observed elevated expressions of HSF1, HSP60, and HSP70 in the aggressively malignant cell lines PC-3, DU-145, and CA-HPV-10. Elevated HSP expression in cancer cell lines appeared to be regulated at the post-messenger ribonucleic acid (mRNA) levels, as indicated by gene chip microarray studies, which indicated little difference in heat shock factor (HSF) or HSP mRNA expression between the normal and malignant prostate cell lines. When we compared the expression patterns of constitutive HSP genes between PC-3 prostate carcinoma cells growing as monolayers in vitro and as tumor xenografts growing in nude mice in vivo, we found a marked reduction in expression of a wide spectrum of the HSPs in PC-3 tumors. This decreased HSP expression pattern in tumors may underlie the increased sensitivity to heat shock of PC-3 tumors. However, the induction by heat shock of HSP genes was not markedly altered by growth in the tumor microenvironment, and HSP40, HSP70, and HSP110 were expressed abundantly after stress in each growth condition. Our experiments indicate therefore that HSF and HSP levels are elevated in the more highly malignant prostate carcinoma cells and also show the dominant nature of the heat shock-induced gene expression, leading to abundant HSP induction in vitro or in vivo. PMID- 15832950 TI - Guidelines for management of asthma: the gaps between theory and practice. PMID- 15832949 TI - Zebrafish Hsp70 is required for embryonic lens formation. AB - Heat shock proteins (Hsps) were originally identified as proteins expressed after exposure of cells to environmental stress. Several Hsps were subsequently shown to play roles as molecular chaperones in normal intracellular protein folding and targeting events and to be expressed during discrete periods in the development of several embryonic tissues. However, only recently have studies begun to address the specific developmental consequences of inhibiting Hsp expression to determine whether these molecular chaperones are required for specific developmental events. We have previously shown that the heat-inducible zebrafish hsp70 gene is expressed during a distinct temporal window of embryonic lens formation at normal growth temperatures. In addition, a 1.5-kb fragment of the zebrafish hsp70 gene promoter is sufficient to direct expression of a gfp reporter gene to the lens, suggesting that the hsp70 gene is expressed as part of the normal lens development program. Here, we used microinjection of morpholino modified antisense oligonucleotides (MOs) to reduce Hsp70 levels during zebrafish development and to show that Hsp70 is required for normal lens formation. Hsp70 MO-injected embryos exhibited a small-eye phenotype relative to wild-type and control-injected animals, with the phenotype discernable during the second day of development. Histological and immunological analysis revealed a small, underdeveloped lens. Numerous terminal deoxynucleotidyl transferase-mediated dUTP fluoroscein nick-end labeling (TUNEL)-positive nuclei appeared in the lens of small-eye embryos after 48 hours postfertilization (hpf), whereas they were no longer apparent in untreated embryos by this age. Lenses transplanted from hsp70 MO-injected embryos into wild-type hosts failed to recover and retained the immature morphology characteristic of the small-eye phenotype, indicating that the lens phenotype is lens autonomous. Our data suggest that the lens defect in hsp70-MO-injected embryos is predominantly at the level of postmitotic lens fiber differentiation, a result supported by the appearance of mature lens organization in these embryos by 5 days postfertilization, once morpholino degradation or dilution has occurred. PMID- 15832951 TI - What do parents of asthmatic children know about asthma?: An Indian perspective. AB - BACKGROUND: Despite the magnitude of the asthma problem, very little is known about the public perception about asthma. METHODS: This descriptive study was conducted to evaluate the knowledge, attitude and practice about the causation, treatment and prognosis of asthma amongst the parents of children with asthma. Subjects were parents of asthmatic children attending two exclusive paediatric hospitals at Chennai, India. A semi-structured pre-validated questionnaire, which included their general understanding on asthma, its triggers and management, was administered. RESULTS: One hundred parents of asthmatic children participated in this study. A diagnosis of asthma was accepted only by 39%, of which only three knew exactly what asthma means. Perception that asthma is contagious was observed by 26%, and 35% believed asthma to be a hereditary disease. Various dietary items were perceived as triggers. Most of the parents (62%) administered oral beta agonist medication at home before proceeding to hospital, but majority were using them as cough medication. Only 13 were administering aerosol therapy at home. Nearly one-third of the parents opined that the disease might remit with advancing age. CONCLUSIONS: General awareness of asthma in the community is poor. Patient education programme should augment awareness, eliminate social stigma and misconcepts in the community regarding asthma. Knowledge about the prevailing perception in the community would be the first step in achieving this. PMID- 15832952 TI - Study on serum and urinary cortisol levels of asthmatic patients after treatment with high dose inhaled beclomethasone dipropionate or budesonide. AB - BACKGROUND: The potential for long-term adverse effects from inhaled corticosteroids relates to their systemic absorption. With increasing use of high dose inhaled corticosteroids, there is need to establish whether similar doses of beclomethasone dipropionate (BDP) and budesonide (BUD) produce clinically important differences in untoward side effects specially hypothalamo-pituitary adrenal (HPA) axis suppression. METHODS: Fifteen asthmatic patients were started on BDP or BUD (2000 microg/day) through spacer for six weeks. Serum cortisol (9 AM and 4 PM), 24-hour urinary steroid and pulmonary function testing parameters were performed. RESULTS: The serum cortisol levels were not found to be suppressed with either BDP or BUD. Similarly no significant changes were found in 24 hours urinary excretion of steroids with either of the drugs. Significant improvement was found in values of forced expiratory volume in the first second (FEV1) with BDP. With BUD the changes in forced vital capacity (FVC) and FEV1 were found to be significant. CONCLUSION: BDP or BUD in high doses of 2000 microg/day given upto six weeks through spacer are equally effective for treatment of bronchial asthma and do not cause any significant change in serum and urinary cortisol levels, and adrenal function/HPA axis. PMID- 15832953 TI - Acute lower respiratory tract infection due to Chlamydia species in children under five years of age. AB - BACKGROUND: The contribution of Chlamydia spp in respiratory tract infections in paediatric population from India has not been studied in detail. METHODS: Sixty children under five years of age who were admitted with acute lower respiratory tract infection during a one year period were investigated for Chlamydial aetiology of respiratory infection. Diagnosis was based on antigen detection by direct immunofluorescence (DIF) in throat swab along with anti-Chlamydial immunoglobulin G (IgG) antibody demonstration by solid phase enzyme immunoassay (EIA). RESULTS: Chlamydia spp antigen was detected in seven (11.6%) cases, C. pneumoniae in six (10%) and C. trachoniatis in one (1.6%). Chlamydia spp IgG antibody in serum was demonstrated in 24 (40%) cases, of which C. pneumoniae IgG was denconstrated in 18 (30%) cases. Taking the criteria of antigen detection (n=7) and high IgG antibody titre of > or = 1:512 (n=5) for a positive case, 12 (20%) children were found to be suffering from recent Chlamydial infection. CONCLUSION: Chlamydia spp plays a significant role in respiratory tract infections in Indian paediatric population. Diagnostic procedure like antigen detection in throat swab is rapid, less cumbersome and feasible and should be more widely used along with antibody demonstration to determine the aetiological agent early in the course of illness. PMID- 15832954 TI - Impact of vascular abnormalities in neoplastic and non-neoplastic lung disease. AB - BACKGROUND: Vascular abnormalities of the pulmonary circulation in the setting of destructive lung diseases caused by inflammation or neoplasia has been scantily researched. A need was felt to document the spectrum of pathological alterations in the vasculature and thus permit speculation into both their pathogenesis and possible clinical significance. METHODS: Between January 1999 and June 2001, 21 patients (male:female 3:4) who had chest disease exceeding a duration of six months and later underwent lobectomy were included in the study. The histopathological material was analysed for vascular changes such as arterial intimal fibrosis, muscularisation of intima and pericapillary fibrosis. The study included a detailed morphometric analysis. RESULTS: The lesions included 15 non neoplastic diseases and six neoplastic diseases. The striking vasculopathic changes observed in the absence of pulmonary hypertension were pulmonary artery medial hypertropy (100%), intimal fibrosis (62%) and muscularisation of the neo intima (3%). Pericapillary fibrosis was seen in 83% of the neoplastic lesions and 67% of the non-neoplastic lesions. CONCLUSION: This study highlights the impact of chronic lung disease on pulmonary vasculature. The role that neoplastic and non-neoplastic lung disease have to play in the evolution of the documented vascular changes have been postulated, and the need to design effective therapeutic strategies to modulate hypoxia, reverse the inflammatory process and stabilise the fibroblastic process is also highlighted. PMID- 15832955 TI - Premenstrual asthma. AB - Gender differences have been recognized in asthma. Specifically in women, an exacerbation in symptoms occurring a few days prior to the onset of menstruation constitutes a phenotype that is not yet fully understood. This phenomenon, called "premenstrual asthma," has been reported to affect upto 40% women with asthma. This article reviews the literature on prevalence, effect of menstrual cycle on symptoms and lung function and discusses the proposed mechanisms of pathogenesis including the effects of female sex hormones on symptoms and beta2 adrenergic receptor function, and the role of airway inflammation. Finally, the various treatment options are presented. PMID- 15832956 TI - Endobronchial non-Hodgkin's lymphoma. AB - A 23-year-old male presented with clinical and radiological features of left lung collapse. Fibre-optic bronchoscopy revealed a smooth globular mass almost completely occluding the left main bronchus. Needle aspiration and endobronchial biopsy from the mass revealed it to be a case of anaplastic large cell lymphoma, a subtype of non-Hodgkin's lymphoma. This report documents the rare presentation of non-Hodgkin's lymphoma as an endobronchial mass. PMID- 15832957 TI - Asymptomatic lymphangitis carcinomatosis due to squamous cell lung carcinoma. AB - Lymphangitis carcinomatosa most commonly due to primary malignancy originating in the breast, stomach, pleura and prostate but may also originate from the lung itself. It is clinically characterised by progressing dyspnoea with or without cough even at an early stage. We report the case of a patient with squamous cell lung cancer presenting with asymptomatic lymphangitis carcinomatosa. PMID- 15832958 TI - Congenital lung hernia. AB - We report a case of a congenital atraumatic lung hernia who underwent a successful operation. PMID- 15832959 TI - Portopulmonary hypertension in a patient of autoimmune hepatitis. AB - A 35-year-old women presented with breathlessness and features suggestive of pulmonary hypertension. Further investigations revealed that she had autoimmune hepatitis and both portal and pulmonary hypertension. Pertinent literature is reviewed. PMID- 15832960 TI - Acromesomelic dysplasia with bronchiectasis. AB - We report a rare case of acromesomelic dysplasia with bilateral bronchiectasis and obstructive sleep apnoea. Diagnosis of acromesomelic dysplasia was based on radiographs of whole skeleton. PMID- 15832961 TI - [New therapeutic strategies in bronchial asthma]. AB - Bronchial asthma becomes one of the most frequent respiratory diseases of the childhood. Progresses of the last decade did not interfere with long term prognosis of the disease. New therapeutic researches try to stop the chronic inflammatory respiratory mechanism: cytokines, leukotrienes, transcription factors, IgE. Some of the new drugs are still only experimental--cytokines antagonists, transcription factors inhibitors, adhesin antibodies, other close to clinical use--new corticosteroids, leukotrienes antagonists, anti IgE antibodies. The new therapeutics methods will be used among the classical drugs and will improve the quality of the patients' life. PMID- 15832962 TI - [Nickel contact dermatitis. A ring signal in actual pathology]. AB - Contact dermatitis produced by nickel is extremely common in women by earrings or other items of jewelry which contain nickel. Areas of involvement are under rings, bracelets, watches, spectacle frames, coins in pockets, jeans studs and other sites of direct contact with the metal. The frequency of nickel dermatitis is increasing in the male population and this may be due of body-piercing or professional contact in the field of metallic constructions. The treatment is difficult because of the presence of nickel in so many substances, things or even food. The management of contact dermatitis include the reduction or elimination of the allergen, the use of topical or systemic steroids, oral desensitisation and use of nickel in selected cases. PMID- 15832963 TI - [HDL-cholesterol--active or passive participant in the pathogenesis of atherosclerosis? (II)]. AB - It is known that high sanguin levels of cholesterol and LDL-cholesterol (LDLc) have an important role in the pathogenesis of atherosclerosis. The treatment of hypercholesterolemia with statins and/or with fibrates have had beneficial effects on coronary heart disease and on other localization of atherosclerosis. The decreased of cholesterol and LDL-cholesterol is the most important effect of this treatment. The epidemiological studies have revealed that the treatment with statins and/or with fibrates produce an increase of HDL-cholesterol (HDLc), which is also very important in the regression of atherosclerosis. We tried in this review to explain the mechanisms of the increase of HDL-cholesterol, in concordance with the data from literature. PMID- 15832964 TI - [New approach in the pharmacologic treatment of cancer]. AB - Carcinogenesis comprises a progressive disorganization of architectural structure as well as a derangement of regulation of normal cell replication and differentiation. This transpires over a variable length of time at each biologic stage. Starting at the molecular level, inherited or acquired errors are associated with alterations in phenotype at successively higher levels of subcellular, cellular and architectural structure and function. Thus the well known "Vogelgram" invokes as a unifying theme the presence of multiple, sequential, functional and structural genetic perturbations that in the aggregate, cause a temporal progression in tumor formation. While these have commonly been thought of as "steps" or "stages" of initiation, promotion and progression, pathologically regarded as the dysplasia or adenocarcinoma sequence, a given series of tumors examined for these changes shows considerable variation in the nature of the manifested genetic changes. Thus progression toward invasive neoplasia is not inexorable, and is opposed by restorative factors that normally operate in healthy and even damaged tissues. Limited damage is recognized and repaired, while cells damaged beyond repair are culled and replaced. As might be expected, such restorative functions perform most effectively early in the mutagenic/carcinogenic process where relatively normal structure and function persist. The evolutionary conflict between these multiple opposing forces likely results in the multi-step and multipath natures described in current models of carcinogenesis. PMID- 15832965 TI - [Balneology, an ancient preventive and therapeutic option in arthrosis]. AB - Arthrosis is defined as a group of distinct diseases with different etiology, having the same biological, morphological, and clinical modifications. The pathological process affects not only the cartilage but also includes the whole joint, the subchondral bone, the capsula, the synovial membrane and the musculature around the joint. Finally, the degenerated cartilage suffers a fibrization, a fissure and an ulceration, disappearing from the surface of the joint. The arthrotic degenerative process is complex and includes numerous factors which initiate the destructive process, progressively altering the joint structure. In fact, the cartilage suffers a chondrolysis process and consequently it disappears from the joint surface. In its turn, the bone reacts by producing a subtendonal osteocondensation type and synovial osteophytosis, responding by an inflammatory reaction, determined by the particles and elements produced by the chondrolysis. The therapy is individualized preferably immediately after the beginning, through non-pharmacological, pharmacological and surgical methods, so that it may influence its evolution and maintain functional the joint touched by the degenerative process. The purpose of the paper is to emphasize the importance of balneotherapy as a non-pharmacological method in the therapy of arthrosis, especially in maintaining the arthrotic joint at a normal level of functionality and of the tissues around the joint, which has a mobilizing and stabilizing function. PMID- 15832966 TI - [Biological and evolutive spectrum of patients ANCA present]. AB - AIM: To analyse the immuno-biological profile of patients with antineutrophil cytoplasmic antibodies (ANCA) presents. This is a retrospective study of all ANCA positives cases from Nephrology Clinic Iasi during five years (1998-2003) on a cohort of 97 patients. RESULTS: pANCA was positive in 28, cANCA was positive in 60 and the others had pANCA and cANCA. The diseases ANCA associated were: systemic vasculitis, rapidly progressive glomerulonephritis (GNRP), chronic renal failure of an unknown etiology. ANCA presence was associated with inflammatory syndrome in 83.5%, anemic syndrome in 71.1%. From the patients with renal disfunction (71.1%), half of them presented an increased level of creatinine (>6 mg%). Renal biopsy proven especially crescentic glomerulonephritis. The complications and the diseases were more frequent in cANCA cases. CONCLUSIONS: cANCA presence is a negative prognosis factor in vasculitis; testing ANCA alone cannot be used to guide treatment. PMID- 15832967 TI - [Chronic myocardial infarction. Clinical symptoms in elderly patients]. AB - Although there is an increasing prevalence of coronary heart disease at the old age, the frequency of chronic myocardial infarction seems to be less in favour of other forms of manifestation. The objective of the study was the assessment of the clinical features of patients with chronic myocardial infarction at a population of coronary patients over 65, admitted in the Department of Cardiac Rehabilitation. METHODS: There have been selected patients over 65 years with ischaemic heart disease: 29 with chronic myocardial infarction (Group A), and 31 with other forms of coronary heart disease (Group B). There were analysed cardiovascular risk factors, symptoms, lipidic profile, echocardiographic findings. RESULTS: Mean age of the groups A and B are 69.9 +/- 4.7 and respectively 71.6 +/- 4.9. Arterial hypertension is more frequent at group B (28/31 vs 22/29, p=0.05); at group A there is a higher rate of smoking (13/29 vs 8/31, p=0.007) and diabetes mellitus (12/29 vs 3/31, p=0.001). Dyspnea is prevalent at group B (26/31 vs 21/29, p=0.01), and the resting heart rate is greater at the same group (89 +/- 19/min vs 74 +/- 14/min, p=0.02). Group A associates more frequent atherosclerotic neurologic and peripheric disease (7/29 vs 5/31 p=0.05 and respectively 6/29 vs 4/31, p=0.01). Although the ejection fraction has similar values (53.9 +/- 10% at group A and 53.8 +/- 14% at group B), the regional contractility abnormalities are more frequent at group A (17/29 vs 11/31, p=0.005). CONCLUSIONS: Old patients with chronic myocardial infarction are especially male gender, associate smoking, diabetes mellitus, other atherosclerotic neurological and peripheric diseases; the regional contractility disturbances are more prevalent at this group. PMID- 15832968 TI - [Relation between arterial compliance and the prematurity of reflected wave on pulse pressure curve evaluated by tonometric sphygmography]. AB - The effect of antihypertensive drugs on the arterial pressure pulse can usually be explained by a change in the timing or intensity of wave reflection or by alteration in the pattern of ventricular ejection. Alteration in timing wave reflection is believed to cause the characteristic changes of pressure wave such: augmentation, augmentation index, "precocity index of second systolic peek" (P2). After calcium channel blockers augmentation decreases from 15 mmHg to 9.4 mmHg, augmentation index P2/P1 (P1 is the first systolic peek) decreases from 132.2 +/- 20% to 129.2 +/- 19.5% and "precocity index of P2" increases from 68.8 +/- 5% ejection duration to 71.8 +/- 4.6% ejection duration. PMID- 15832969 TI - [Acute myocardial infarction with neurological symptoms: clinical and pathologic aspects]. AB - A variety neurological symptoms can be the onset of an acute myocardial infarction. The study includes 96 patients who have undergone necropsies (from 1978 to 2003), hospitalized at the "Sfanta Treime" University Hospital, Iasi, in the Neurology Clinic. They were admitted for various neurological manifestations: hemiparesis, hemiplegia, aphasia, coma of the 1st degree up to the 4th. Death of these patients was due to acute myocardial infarction in the cases of most of them, even those with significant brain damage. We have taken into account the anatomic pathology of the heart, brain, kidneys and lungs. The prevalence of the factors of cardiovascular risk was considered in our study, as well as the topography of the myocardial infarction. Patients with acute myocardial infarction can present major neurological symptoms with no significant cardiovascular clinical symptoms. PMID- 15832970 TI - [Radiofrequency treatment of cardiac arrhythmias at the Cardiology Center in Iasi ]. AB - It is well known that the radiofrequency catheter ablation (RFCA) treatment of cardiac arrhythmias is effective and devoid of significant complications. However, in our country, the experience of this technique is not so large and requires more clinical studies to validate it. OBJECTIVES: The aim of this was to present our experience in the RFCA of cardiac tachycardias. METHODS: Between July 2002 and February 2003, in Iasi Cardiology Center, we performed a total of 79 diagnosis procedures. Of these 57 (72%) were radiofrequency ablation procedures at 51 patients, mean age 48 +/- 16 years (12-81), men majority 32 (63%). The electrophysiology diagnosis was: 22 (39%) common atrial flutter, 3 (5%) atrial tachycardias, 13 (23%) atrioventricular nodal reentrant tachycardia, 12 (21%) Wolff-Parkinson-White syndrome, 4 (7%) orthodromic tachycardia using concealed accessory pathway, 2 (3%) ventricular tachycardias and 1 (2%) ablation of the atrioventricular junction. RESULTS: The initial ablation procedure was effective in 44/ 51 patients (86%), with a total duration of 130 +/- 42 min, using 24 +/- 10 min of radioscopy. During follow-up 13 +/- 1.3 (11-16 months), recurrence occurred in 7 patients 14%. In 7 of them a second effective procedure was performed in 4 patients. Finally, radiofrequency ablation was effective in 48/51 of patients (94%), improving the procedural success rate at 95% (54/57 patients). No complications occurred during follow-up. CONCLUSION: The treatment of cardiac arrhythmias by radiofrequency catheter ablation is effective and safe and is the only way to cure the supraventricular and ventricular tachyarrhythmias. PMID- 15832971 TI - [Respiratory syndromes in acute poisoning]. AB - Respiratory syndromes in acute poisoning can refer to a wide range of specific clinical syndromes, from acute tracheobronchitis to acute pulmonary edema, chemical pneumonia, acute respiratory distress syndrome and respiratory failure, that occur as a result of direct or indirect effect of chemical substances, drugs and toxins on lungs and airways. Our study attempt to identify, during one-year retrospective study on patients diagnosed with acute poisoning, addressed to Medical Clinic of Emergency Clinic Hospital of Iasi, the respiratory syndromes commonly associated with acute poisoning. We found that the association of toxins, inhalation of gases or volatile substances have a high risk for appearance of respiratory syndromes. The outcome of these patients is influenced by the duration of exposure or the delay of presentation to the hospital after ingestion, and depends on the rapid and aggressive measures for basic life support and intensive care. PMID- 15832972 TI - [The study of two immunological markers in patients with type-1 diabetes and in their first degree relatives]. AB - The prediction of diabetes mellitus is mostly based on the existence of plasma markers. The aim of this preliminary study was to determine islet cell antibodies (ICA) and glutamic acid decarboxylase antibodies (GADA) in 28 diabetic children (12 of them having an evolutive disease of 1 year and 16 at the beginning of the diabetes) and to 47 of their first-degree relatives. There have been determined the levels of these two autoantibodies using the ELISA technique. RESULTS: To 17 of the patients with type I diabetes have been found high levels of GADA (60.7%) while 8 cases have positive ICA (28.5%). For the patients whose disease was diagnosed 1 year ago there have been found differences between the patients with and without antibodies regarding the level of the average values of Hb A1c, the daily insulin needs and the remission period. From the tested parents (a total of 25), 7 was GADA positive (28%), 6 had both antibodies present (24%) and one mother was ICA positive (4%). 9 of the brothers and sisters of the diabetic patients had high levels of GADA and 2 had both antibodies present. To the first degree relatives with autoantibodies must be determined other plasma markers too (IAA, IA-2A) as well as genetic markers (HLA typing). CONCLUSION: The use of plasma markers is recommended as a first step in discovering the relatives with potential risk of developing the disease. PMID- 15832973 TI - [Lipids profile in a group of hypothyroid patients vs. treated hypothyroid patients]. AB - Hypothyroidism, like obesity, is one of the pathological conditions most frequently associated to disorders of the lipid metabolism. Hypercholesterolemia is favored, due to the hormone deficit and to the decreased activity of the lipoprotein lipase. MATERIALS AND METHOD: We observed the determination of the serum concentrations of TSH, of the total cholesterol level, of the triglycerides, HDL, LDL, VLDL and BMI as well. RESULTS AND DISCUSSIONS: We observed the predominance of the female hypothyroid patients, ages 40 through 60 and BMI > 30kg/m2. The presence of the heart condition in the studied group was the predominance of the cases of ischemic heart disease. The cholesterol levels varied depending on the TSH levels. There are multiple mechanisms of the dyslipemic syndrome, its intensity depending on the degree of the thyroid insufficiency, however, not influenced being by its cause. It can be considered an indicator of the disease evolution or of the efficiency of the substitution treatment strategy. The absence of the total normalization of the lipid metabolism, under the conditions of a correct treatment strategy, may raise suspicions regarding a possible associated primitive hyperlipidemia, who need typical therapy. PMID- 15832974 TI - [Prospective epidemiologic and clinical observations on the prevalence of risk factors for stroke, performed on 374 inpatients, during the period of June 2001 June 2004]. AB - Within the framework of cardiovascular pathology, with an epidemopandemic aspect, the stroke, through its mortality and sequels, represents, worldwide, a major problem of community health. In this context, we present the results of an epidemiological and clinical study, both prospective and interdisciplinary, performed on a sample of 374 inpatients suffering from stroke, hospitalized in the Neurology Ist Clinic of Iasi, during the period June 2001-June 2004. The characteristics of the studied sample: age--78.8% over 51 years old, sex--53.7% male; residence--51.4% urban; provenance: rural--70.0% (p<0.001); studies: higher education--7.7% ; pensioners--76.2%; passive finding--96.8%; previous hospitalizations for cardiovascular diseases and/or diabetes mellitus: 48.2%; blood groups: OI and AII--86.61%. RESULTS: Risk factors for stroke: 56.6%--BMI > 25; "A" behavior type (Bortner's test)--63.8%; stress: familial--74.1% and occupational--25.9%; excess consumption: alcohol--11.4%; salt--32.8%; tobacco- 24.9%; passive smoking--10.8%. Releasing and aggravating risk factors: arterial hypertension (AHT)--55.4% (p<0.001); obesity--12.9%; diabetes mellitus--12.5%. Hereditary antecedents: AHT--46.8%; ischemic cardiopathy and myocardial infarction--15.6%; obesity and dyslipidemia--15.0%; stroke--10.8%. CONCLUSIONS: In this study, the multifactorial and associative etiology of stroke is confirmed once more, and the estimate costs of 1.21 milliard lei, only for the 10-day average hospitalization, for each of the 374 inpatients. PMID- 15832975 TI - [Clinical and developmental aspects of cardiac involvement in infant of diabetic mother]. AB - The authors evaluated the main clinical and evolutive aspects of heart involvement in the newborn of diabetic mother (IDM). We studied the files of 35 IDM in a 4 years period; they were investigated the first week of life and at 6 12 months (18 cases). Most of the patients were asymptomatic/signs of extracardiac suffering; 15 had a systolic murmur. ECG: left ventricle (LV) hypertrophy (8) and LV repolarization disturbances (19 cases). Chest X-ray: cardiomegaly (9 cases). Echocardiography: hypertrophic cardiomyopathy (HCM), especially septal (25 cases, 71%); LV diastolic dysfunction (19-35) and normal systolic function; pulmonary hypertension (3), other congenital heart diseases (8 cases). The control revealed the alleviation of the LV walls size and LV diastolic function. The high incidence of the cardiac manifestations in IDM and the risk of occurrence of some severe problems, require a complete cardiological exam from the first few days of life and a follow-up schedule until the normalization of the cardiac parameters. PMID- 15832976 TI - [Acute renal failure in children. Study of 35 patients]. AB - The medical care of children with acute renal failure and the necessity of the substituting the renal functions has dramatically modified over the past 15 years. The study has been conducted retrospectively on 35 children diagnosed with acute renal failure (ARF), for analyzing the etiological spectrum, the evolutionary patterns and the influence factors of the ARF evolution. The following parameters have been taken into consideration: ARF etiology, hTA/HTA, oliguria, level of serum creatinine, the type of treatment (renal substitution by hemodialysis, peritoneal dialysis, or conservative treatment). The ARF etiology is dominated by the sepsis (31.4%) and by the hemolytic and uremic syndrome (17.1%). The treatment applied was conservative in 48.5% of the cases; 51.50% of the patients were in critical state and required extrarenal substitution by hemodialysis--42.8%--and peritoneal dialysis--8.5%. Global mortality is reduced (22.8%), but in the dialysed patients it is of 44.44%. The main death causes were: severe hepatic failure, oncological diseases, severe neurological damage and hemodynamic damages. In conclusion, the ARF prognosis in children is influenced by the comorbidity states. Dialysis has improved the therapeutic results in the currently presented lot, the death causes being extrarenal. PMID- 15832977 TI - [Neurocutaneous syndromes and epilepsy. Report of 23 cases and review of the literature]. AB - The neurocutaneous syndromes are characterized by congenital dysplastic abnormalities involving the skin and nervous system. We included in the study 23 children diagnosed with different types of neurocutaneous syndrome (10 patients with tuberous sclerosis, 5 with Sturge-Weber syndrome and 8 with neurofibromatosis type 1 and epilepsy'. The factors studied included: age at onset of seizures, seizure types and frequency, seizure response of treatment, EEG, CT/MRI, neurologic, neuro-ophthalmologic and psychological examination. In 39.13% of cases the epilepsy onset preceded the clinical diagnosis of neurocutaneous disorders. The age of seizure onset ranged from 4 month to 5 years. The most common type of seizure appear to be complex partial. 21.73% of patients associated mental retardation. Therapeutical efficacy was variable depending on type of seizure and type of neurocutaneous syndromes. PMID- 15832978 TI - [Use of simulator in treatment planning and determination and definition of treatment volume]. AB - Achieving optimum results in radiation therapy depends on delivering a homogeneous, tumoricidal dose to a well-defined region while minimizing the dose to surrounding normal tissue. Accomplishing this task requires a high degree of accuracy in dose delivery along with a systematic and logical approach to the treatment of the particular disease. In attaining this goal, the use of the simulator is absolutely essential. PMID- 15832979 TI - Diagnostic traps in porphyria: case report and literature review. AB - Porphyrias are metabolic disorders of heme biosynthesis, which encompass a broad range of symptoms and signs, neurologic, cutaneous or mixed. Because of lack of specificity and polymorphous clinical picture, porphyrias can mimic either neuropsychiatric, dermatologic, or gastrointestinal diseases. We present the case of a 58 years old man to whom clinical presentation suspicious of Addison's disease (melanoderma, fatigue, weight loss, intermittent abdominal pain) was the disguise of porphyria cutanea tarda. A general background of porphyrias and differential diagnosis with other forms of hepatic porphyria, as well as other causes of hyperpigmentation, are given. The clinician should be aware of the protean manifestations of porphyrias and include them in clinical judgment in various situations. PMID- 15832980 TI - [Arrhythmic cardiomyopathy. Case report]. AB - An 11 year-old boy was admitted with incessant sinus node reentrant tachycardia and secondary dilated arrhythmic cardiomyopathy, treated by radiofrequency ablation. Two years later he was admitted with incessant automatic atrial tachycardia and arrhythmic cardiomyopathy; a second catheter ablation procedure failed, but the third one, performed four month later, was successfully and resulted in a restoration of a normal sinus rhythm and a complete regression of arrhythmic cardiomyopathy. PMID- 15832981 TI - [Melanotic neurofibroma, clinical and histopathologic diagnosis. Case report]. AB - Melanotic neurofibroma is a rare benign tumor, derived from peripheral nerve sheath, whose originality consists in the presence of melanic pigment. The clinical diagnosis is difficult to establish, requiring the histopathological exam to make the difference between melanotic neurofibroma and the other pigmented tumors. Although, sometimes neither the anatomopathological exam can establish the final diagnosis, requiring supplementary studies. Melanotic neurofibroma has a good prognosis and the malignization is rare. The elective treatment is surgical, represented by the complete excision of the tumor. PMID- 15832982 TI - [Mixed tumour of submandibular salivary gland. Case report]. AB - The authors present the case of 14 years male child with tumour located on submandibular salivary glands. It was proceeded the biopsy and tumour excision, the tissue fragments being further processed at paraffin, sectioned and than stained HE, PAS, Alcian-Blue, Van Gieson and Gordon-Sweet. The first biopsy performed from the latero-cervical ganglion revealed the presence of an benign tumour of salivary gland. Totally excision of the tumour emphasized the presence of a salivary gland encapsulated tumour, sized 2.5/2.5/2 cm, nodule shaped, white colored, hard consistency. Histopathologic examination revealed the existence of a proliferating encapsulated tumor, well separated from the normal adjacent tissue. The small sized tumour cells with moderate cytoplasm induce formation of glandular lumens, some of them with cystic dilatation, with mucous content. Other tumour cells form small cords or nests. The tumour stroma forms mucoid areas, some with osteoid appearance. We have presented a case of a 14 years aged child with pleomorphic adenoma with rare location within the submandibular salivary gland. The post biopsy rapid increase of the tumour imposed the totally surgical gland excision. PMID- 15832983 TI - [Spontaneous intracerebral haemorrhage. An update of conservative treatment]. AB - In our article we analysed the current conservative therapeutic options in spontaneous intracerebral haemorrhage as well as a few controversies regarding the efficiency of some types of treatment. Spontaneous intracerebral haemorrhage represents a major neurosurgical emergency. In spite of the severe prognosis of the disease, there is reason for optimism. There are important improvements in treatment of arterial hypertension, and cerebral lesions. The aim of the conservative medical treatment is to maintain good oxygenation, decreased arterial pressure, control of the intracranial hypertension, cerebral oedema, prevention of seizures. All these contribute to improving the vital and functional prognosis. PMID- 15832984 TI - [Surgical treatment of Graves' disease]. AB - The surgical treatment of the Graves' disease is no longer a final solution but the main option, as for all the benign thyroid diseases. Between January 1994 and December 2003 in the 3rd Surgical Unit of the Sf. Spiridon Hospital of Iasi 805 operations were performed for benign thyroid disorders, 78 of which were performed for the Graves' disease. All the patients had been investigated and diagnosed in the Endocrinology Unit of the same hospital, where the necessary medical treatments had been administered. The patients transferred to Surgery had a medium age of 38 (extremes 17-79 years). Women had a greater proportion, as expected (sex ratio F: M = 4.57:1). Some patient had concurrent diseases (5 cases with atrial fibrillation and 3 with chronic hypocalcaemia). 58 total and 20 subtotal thyroidectomy were performed. There were few immediate (3 bleedings which needed reoperation for haemostasis) and late complications (2 cases of postoperative hypocalcaemia which received medical treatment). 4 relapses after previously performed subtotal thyroidectomy were diagnosed and treated. Total thyroidectomy is the treatment of choice for the Graves' disease. The frequency of complications is similar to that occurred after subtotal thyroidectomy, the functional results are immediate, predictable and stable, and the substitutive treatment is very efficient and easy to administer. PMID- 15832985 TI - [Treatment of non-arterial ischemic anterior optic neuropathy. Clinical study]. AB - The study was performed on 53 patients diagnosed with non arterial anterior optic neuropathy treated with dopamine. After a monitoring period of 5 years it seems that an increase in visual acuity at these patients occurred. The main conclusion is that using dopamine in some cases of non arterial anterior optic neuropathy is necessary and beneficial. PMID- 15832986 TI - [Chlamydia infections and tubal infertility]. AB - This is a screening study for chlamydial infections (1153 patients females). In the number of 272 cases (23.59%) chlamydial infection was confirmed by ELISA blood tests. We purpose to determine the risk factors for tubal infertility, because Chlamydia trachomatis (CT) is the most important cause of pelvic inflammatory disease (PID) and permanent tubal damage. High correlations (correlation coefficient-"r") of chlamydial infection with hygienic level (r=0.51), avoided use of condoms (r=0.58) or number of abortions (r=0.65) were noticed. Results with primary prevention through sexual education have been disappointing, so is necessary preventing or limiting sequelae of chlamydial infection, such as PID and tubal infertility. PMID- 15832987 TI - [Complications of intracerebral hemorrhage caused by arterial hypertension]. AB - Intracerebral hematoma represent a frequent complication of hypertension. It is accompanied by significant mortality and complications. It is mainly a surgical condition but we prefer conservative therapeutic options in cases where feasible. The conservative treatment gives better results but surgery becomes obsolete in cases when medical treatment fails. PMID- 15832988 TI - [Cervugid ovules in cervico-vaginal infections and cervix uteri precancerous conditions treatment]. AB - This medicine was authorized by the National Drug Agency (ANM, Bucuresti) in 2001. PURPOSE: To evaluate the effectiveness and the tolerance to Cervugid ovules, a preparation that combines the polyvalent local antiinflammatory action of chloramphenicol, metronidazole and nystatin with the effect of hydrocortisone acetate, an unspecific anti-inflammatory agent; they all are embedded in a Lipex 403, semisynthetic fat. MATERIALS AND METHODS: The evaluation of 500 patients ages between 15 and 85 years with genital infections, registered in the files of "Cl. II Obst. and Gynecology" of the Cuza-Voda Hospital from Iasi has been studied. We studied the subjective manifestations (local discomfort and pelvic pains, local burning and dryness,vulvovaginal itching and dyspareunia) and objective manifestations (vaginal and cervical secretion, the cytotest performed and colored though the Papanicolaou method and reported in the Bethesda system). RESULTS AND DISCUSSION: Healing of the subjective symptoms in 98%, healing of the leukorrhea--as a main objective symptom--in 95%; The Bethesda system cytotest was one of the inflammatory type in the most of the cases and there wew found in 85 cases: 6 ASCUS, 41 LSIL, and 37 HSIL. The use of Cervugid had a healing response in most of the cases when used in acute and chronic cervico-vaginal inflammatory processes. Cervugid may be considered as an important agent in the treatment of the precancerous affections af the cervix uteri on the following reasons: zhe cure of the infections caused by chlamydia, involved in the etiology of cervical neoplasms, the cure of the HPV infection under episome form, classified in the Bethesda system within the ASCUS, AGUS or LSIL classes. When the cytotest was in the HSIL class, a conization in the LLETZ method was performed. CONCLUSIONS: Cervugid is conceived for those three main categories of pathogenic factors related to the etiology of cervico-vaginitis: microbia germs, protozoa and mycosis. In addition, it is active on chlamydia and mycoplasms, always sensitive to chloramphenicol therapy. That is why Cervugid with in local administration is indicated in the microbial, trichomoniasis and mycotic vaginitis caused by one category of pathogenic agents or by associated forms, in cervicitis, in the pelvic inflammatory processes (pelvic congestion, metritis, adnexitis, and inflammatory processes associated with benign or malignant tumors of the genital apparatus). The results obtained proved that Cervugid is highly effective medicine. PMID- 15832989 TI - [Synchronous sigmoid and superior rectal resection and left hepatic lobectomy extended to Spiegel lobe for a sigmoid cancer with hepatic metastasis. Case report]. AB - The recent developments of surgical technologies allowed the achievement of some standardized interventions with anatomical and functional visa, which based on the improvement of anesthesia and intensive care, and not least by elaboration of efficient chemotherapy protocols, determined new horizons in the treatment of advanced cancers. This work presents a case witch was hospitalized at the Department of Hepatic Surgery, of City Hospital from Timisoara for a colorectal cancer stage IV (T3N1M1), with hepatic metastasis localized at the left hepatic lobe (II and III segments) and Spiegel lobe. A surgical intervention was performed, when in the same operating time was practiced a sigmoid and superior rectal resection (Hartmann) and also a left hepatic lobotomy extended to the first segment. The post operating evolution of the patient was favorable and also after fourth month from the surgery, when no signs of relapse were established at reevaluation. PMID- 15832990 TI - [Actinomycosis of the appendix. Case report]. AB - Abdominal actinomycosis is a chronic granulomatous suppurative disease caused by anaerobic Gram positive germs from Actinomyces species. The vermiform appendix and ileocaecal region have been recognized as the most commonly involved sites. Actinomycosis of the appendix is generally acute in presentation, mimicking a common acute appendicitis. We present a case of actinomycosis of the appendix in a 62-year-old male patient. The clinical presentation was as for a common acute appendicitis. The ultrasound examination sustained the diagnosis of acute appendicitis and an appendectomy was performed. The diagnosis was established by histo-pathologic examination. The cultures from intraperitoneal fluid were negative. The association of a long-standing antibiotic therapy to the surgical treatment resulted in definitive cure for this patient. Actinomycosis of the appendix is a rare disease that must be known, the diagnostic being the key for a successful treatment. PMID- 15832991 TI - [Spontaneous rupture of the esophagus. Case report]. AB - We present a 50 years old man who was admitted for severe pain in the chest appeared after vomiting, dyspnoea, cardiovascular collapse. The diagnosis of spontaneous perforation of the esophagus (Boerhaave's syndrome) was confirmed by a radiopaque swallow. We performed a laparotomy, mediastinal drainage, cervical esophagostomy and jejunostomy. Postoperatively, the general state was severe- with high fever, important pleurezia, necessitating pleural drainage, mechanical ventilation for 8 days, parenteral and jejunostomy nutrition. It was possible to close the esophagostomy in the 43rd postoperative day. The patient has been dismissed on the 59th day. Five days later he presented in the ER with severe dyspnoea due to tracheal stenosis--emergency tracheotomy was performed, followed by resection of the stenotic cartilages. PMID- 15832992 TI - ["Mammary loop" technique--new method of multiple coronary revascularization using internal mammary arteries]. AB - The internal mammary arteries are "the gold standard" conduits for coronary by pass because their' s patency is superior to the saphenous graft. There are some surgical techniques which allow to use the entire length of the mammary arteries: the Y graft, the T graft and the pi graft. We describe recently the "mammary loop" technique that allows to make a Y graft using only one internal mammary artery. This technique also simplifies the construction of a pi graft. In this article we present the variants of this technique and discuss the indications and our early experience results. PMID- 15832993 TI - Interactions between extracellular and intracellular administered angiotensin II in isolated rat portal vein rings. AB - It is already demonstrated that intracellular angiotensin II (Ang II) stimulates smooth muscle contraction and cell growth. We studied the contractile effects of Ang II intracellular delivered by the means of liposomes (LAngII) and also the interactions between intracellular and extracellular administered Ang II on the rat portal vein rings without endothelium. The results were expressed as the percentages of the control contraction (K+ 40 mM; mean +/- S.E.M). LAngII induced contractions were of 120.46 +/- 8.06%. On the other hand, 0.1 microM Ang II produced contractions of 121.43 +/- 6.83%. Both Ang II and losartan, administered either extracellular in the organ bath or intracellular by the means of liposomes, inhibited the contractions induced by intracellular Ang II. The inhibitory effects of losartan on LAngII-induced contractions were dose dependent. Thus, 10 microM losartan strongly blocked (10.01 +/- 1.41%) and 1 microM losartan partially blocked (39.73 +/- 5.35%) the LAngII-induced effects. LLOS significantly inhibited the LAngII contractile effects (38.51 +/- 8.92%). Our results revealed that LAngII partially inhibited the contractions induced by extracellular administered Ang II (42.42 +/- 3.29%). On the other hand, 0.1 microM Ang II also inhibited the contractile effects induced by LAngII (67.42 +/- 0.76%), although a little bit less. So, contractions induced by Ang II administered intracellular are mainly mediated by intracellular Ang II receptors sensitive to losartan. At the same time, the participation of cell membrane angiotensin receptors to intracellular Ang II effects cannot be excluded. PMID- 15832994 TI - [Evaluation of the angiogenic process in breast carcinoma--correlations with cancer aggressiveness]. AB - Angiogenesis represents an essential event required by tumors to support their growth. The aim of our study was to investigate the presence of neovascularization in 44 primary breast carcinomas and in 24 axillary lymph nodes metastases and to establish a possible correlation between the presence of tumor angiogenesis, some clinical and pathological features of the cases and the expression of p53, an important cell cycle regulator. To identify the new blood vessels, we used immunohistochemistry for von Willebrand factor, a marker of the endothelial cells. The results showed that 77.27% of the primary breast carcinomas and 75% of the lymph nodes metastases are positive for von Willebrand factor and this positivity is significantly correlated (p < 0.05) with the expression of p53, supporting the idea that angiogenesis is a marker for tumor aggressiveness and p53 could be involved in this process. PMID- 15832995 TI - [Prognostic markers in gastric neuro-endocrine carcinoma]. AB - Gastric endocrine tumors show a wide spectrum of clinical pathologic features. Behaviour prognostic evaluation of these tumors is difficult. The aim of this report is to present a rare case of a neuroendocrine carcinoma with multiple locations in stomach. METHODS: A 62 years female patient underwent a resection of stomach for a tumor. The paraffin-embedded sections of 12 blocks were studied by histological and immunohistochemical methods including HE, van Gieson, Grimelius, chromogranin A, cytokeratin, neuron-specific enolase, vimentin, desmin, smooth muscle actin, Ki-67 antigen, protein S-100. RESULTS: Histological pattern was similar in primary tumor (mucosa and submucosa) and sessile lesion at distance, where it affected only mucosa. Histopathologically, these tumors show a sheet of round to spindle and polyhedral neoplastic cells with hyperchromatic nuclei, considerable mitotic activity. Immunohistochemically, the neoplastic cells were positive for cytokeratin and neuron-specific enolase and a high Ki-67 labelling index. The three lymph nodes from five had metastasis. CONCLUSION: The IHC reactions were needed for diagnosis of multiple neuroendocrine carcinomas with lymph node metastasis. These histological and immunohistochemical criteria were very important for the identification of tumors with the aggressive evolution. PMID- 15832996 TI - [Perilesional changes in gastric ulcer]. AB - Our work aims to provide a descriptive analysis of the perilesional changes associated with gastric ulcer, which were observed on a group of patients diagnosed by endobiopsy. MATERIAL AND METHOD: The group comprised 90 persons with clinical diagnosis of gastric ulcer, who were investigated both endoscopically and pathologically. The gastric fragments were processed for light microscopy (HE, PAS, alcian blue stains). RESULTS: Besides the typical gastric ulcer, the morphological exam identified gastric ulcer combined with chronic gastritis, complete and incomplete intestinal metaplasia, gastric atrophy, hyperplasia as well as dysplasia. A malignant gastric ulcer was confirmed in 6 cases. A number of 10 patients displayed malignant lesions (gastric carcinoma, intestinal and diffuse types) developed independently of the gastric ulcer. CONCLUSIONS: The identification of various morphological changes in the gastric mucosa adjacent to the ulcer allows a correct evaluation, with respect to the diagnosis and the evolution. The presence of these lesions, correlated with the existence of Helicobacter pylori, means major risk factors for gastric carcinogenesis. PMID- 15832997 TI - [Drug politics--economic view-point of a practitioner]. AB - Any expenditure made in a domain leads to the diminution of the expenditures in another domain. The list of reimbursed drugs should be decided after a cost effectiveness analysis. The treatments considered too expensive in the developed countries are even more expensive for a country like Romania. The practice guidelines do not consider the economic analysis of the treatments. As soon as the results of a study are statistically significant, the specialty societies include that drug into their guidelines. A treatment is more cost effective when the risk is higher, therefore the reimbursement should have to be done function of risk, too. PMID- 15832998 TI - Assessment of some performance indicators of medical departments in tertiary hospitals. AB - The present study assesses the performance of 37 medical departments from 12 tertiary hospitals in Iasi town during year 2001 through the following indicators: average hospital stay, utilization rate and hospital bed turnover rate. The first indicator has been analyzed by comparing the values recorded in various medical departments with those recommended by the Ministry of Health for tertiary hospitals. The highest value was recorded in the nephrology department, with a ratio 2001 value/optimal value of 2.76. Long average hospital stay has been recorded in 14 of the 37 assessed departments (37.8%). During year 2001, the following utilization rates have been found: normal rates of 300-365 inpatient days per hospital bed in 14 departments, high rates (over 365 days) in 11 departments, and low rates (less than 300) in 12 departments. Bed turnover rate varied with department's specialty profile and length of hospital stay from 11 inpatients per hospital bed in the acute mental disorders department to 146 in C intensive care unit. PMID- 15832999 TI - [Evaluation of the effectiveness and limits of balneology therapy for arthrosis in Lacu Sarat Health Resort, Braila, Romania, in the period of 1994-2003]. AB - This research tried to estimate the importance of the precocious diagnosis in the arthrosis. It also analyzed the efficiency and limits of the balneology therapy based on the facilities offered by Lacu Sarat Health Resort, Braila. The approaching method consisted of a retrospective analysis of 10565 standard records of patients suffering from arthrosis, hospitalized in the health resort, between 1994-2003. The cases fell into various categories, according to the illnesses, as follows: 2854 (27.01%) gonarthrosis, 1247 (11.82%) coxarthrosis, 2001 (18.91%) spondylosis, 2003 (18.87%) peripheral arthrosis. The balneology treatment was applied individually, taking into consideration the location, clinical stage and associated illnesses. The treatment also included physiotherapy and kinetic therapy, adjusted to the stage of disease. The treatment results were good in 73.30% of cases. The patients' recovery aims at stabilizing the development of the illness, removing the objective as well as subjective suffering and maintaining the mobility and ability of the affected segment. The conclusions of the research underline the particular contribution of the clinical evaluation in diagnosis and the efficiency of external balneology in removing the pain and inflammation, maintaining the articular mobility and ability. Thus, the cure can ensure an appropriate articular comfort for most of the patients, taking into consideration the recurrent character of the illness, with repeated periods of suffering and pain. PMID- 15833000 TI - [Dento-stomato-facial rehabilitation of young patients. Practical alternatives]. AB - The design conception and the technological achievement of every gnato-prosthetic device are conditioned by a minute evaluation of the biological and morphological individualities of each case, the social level of the patient, corroborated with the technological supply. In our study we evaluated different modalities of dento stomato-facial rehabilitation of young patients with partially reduced maxillary or mandibular dentition. The study was carried out on a group of 12 young patients, 5 males and 7 females, aged between 16-24 years, who presented at the clinic for the treatment of complex functional disorders that occurred as a consequence of the dentition. The fixed mixed gnatoprosthetic therapy for the young patients and teenagers has to be adapted to the surveyed clinical individuality. The success of the therapy is assured by the accurate application of the gnatoprosthetic principles, the accurate achievement of the technological steps, the use of compatible biomaterials according to the dento-stomato-facial features of this age category. PMID- 15833001 TI - [Infection control in dentistry. Educational requirements]. AB - The need for an effective infection control program has always been an essential and integral part of the dental practice. Despite human and material efforts, occupational diseases continue to consume significant health care worldwide and the risk of "cross-infection" remains high. The European dental education requires the specialists in this field to have a high level of medical training, the standards for clinical skills and abilities including knowledge on the control of "cross infection" and the prevention of physical, chemical, and microbial contamination. The results of a questionnaire-based assessment of the knowledge gained by the last year students (VI) of the Iasi School of Dentistry revealed the need for improving their knowledge in this field by inducing firm clinical behavioral abilities, organization of practical "exercises" in ideal circumstances and conditions for applying the infection control protocols. PMID- 15833002 TI - [Simulation models of the processes of the interface in dental structures]. AB - The occlusal forces have an effect on the dental structure in the longitudinal axis of the tooth, on the one hand, but, on the other hand, in the horizontal plane, they are conducted through interdental contact points. In this paper we try to simulate the stress and the deformations determined by different forces in different directions, over the dental interfaces. We have used the method of finite elements and Fastflo domain for the mathematical calculations of probability. Our conclusions are that the changes in the deformation field appear only for those materials that comply with the rate of elasticity changes in the condition where delta = E1/E2 < 20. PMID- 15833003 TI - [The amount of nitrites in edible mushrooms harvested from Bistrita Valley]. AB - In our study has been determined the amount of nitrites in edible mushrooms which grown in different zones from Bistrita Valley, near to the "Fibrex-nylon" Complex Savinesti. The nitrites has been extracted with water, in the aqueous extract we determined spectrophotometrically nitrites with Griesz Reagent. The results were expressed in mg nitrites/g dried product (105 degrees C). We analysed 41 samples of edible mushrooms (22 samples harvested on october 2001, 19 harvested on september 2002). The nitrites concentrations in mushrooms are between 2.06 and 49.88 (samples harvested on october 2001), between 1.98 and 31.84 (samples harvested on september 2002). The highest values for samples harvested on october 2001 was determined in Laccaria lacata, Collybia butiracea, Hygrophorus chrysodon, Russula nigricans, for the samples harvested on september 2002 the highest values was determined in Boletus granulatus, Macrolepiota procera. PMID- 15833004 TI - [Qualitative determination of flavonoids and phenol-carboxylic acids of indigene Stachys species using HPLC method]. AB - Indigene Stachys species have a high content in interesting compounds as iridoids, but also high quantities of flavonoids and phenol-carboxylic acids. These compounds present vasorelaxing effect on smooth vascular muscle interfering with metabolic degradation of nitric oxide pathways (inhibition of peroxynitrites formation) and by inhibition of phosphodiesterase. This work shows flavonoids spectrum of 3 Stachys species using HPLC methods. We identified quercetol, rutoside, miricetin and phenol-carboxylic acids (chlorogenic, caffeic and ferulic acids). PMID- 15833005 TI - [Means of purification of cephalexin with a view to therapeutic use]. AB - Cephalexin (CEX) is the generic word of chemical compound 7-(D-alpha aminophenylacetamido)-3-methyl-3-cephem-4-carboxylic acid, which is part of first generation oral cephalosporins group. It is use as cephalexin monohydrochloride monohydrate (CEX.HCl.H2O) syrup or tablets for oral administration. For obtaining a compound with great solubility and osmotic pressure, which is ideal for pharmaceutical forms with controlled dosage, it is necessary to achieve the crystalline form of Cephalexin monohydrochloride monohydrate. The therapeutic use Cephalexin has an output over 95 %, so it is necessary to purify CEX depending on it's isoelectric pH (pKa), which is 4.2. A good purification took place at greater or less values of pH, then the isoelectric pH value of CEX. The purification of CEX at greater values of pH then isoelectric point, took place with a greater output (75-76%), and the obtaining Cephalexin is much pure (97 98%). PMID- 15833006 TI - Drugs and the QT interval--implications for dentistry. PMID- 15833007 TI - Pretreatment wax-ups and provisionals for restorative dentistry. AB - One of the most important factors in restorative success is the preparation of a case prior to treatment. Case planning, constructing the wax-up and provisional restorations, and understanding the potential problems with a case prior to beginning can only ensure a higher degree of success than would be achieved by not employing these basic restorative steps. Although it is not utilized generally, the wax-up can be of immense value to any restorative dentist for cases of all kinds, ranging from a single tooth to full-mouth reconstruction. PMID- 15833008 TI - Time and boundaries. PMID- 15833009 TI - Are bleaching lights the real deal? PMID- 15833010 TI - Identify, protect, restore: emerging issues in approaching children's oral health. AB - Treating dental caries as an infectious disease requires the clinician to identify risk factors and tooth demineralization as early as possible. This early recognition of risk allows the dentist to implement prevention regimens before significant oral disease becomes apparent. Prevention includes rebalancing the oral environment to its natural state to prevent oral disease progression. Bacterial control, buffering acidic pH, and providing calcium, phosphate, and fluoride all can aid in rebalancing the oral cavity. Rebalancing can protect potential demineralization of tooth structure; it also can repair early lesions. Once cavitation occurs, conservative restoration with biomimetic materials is indicated. This approach dictates identifying and assessing potential caries risk factors early, preventing caries by addressing these primary risk factors, eliminating or minimizing their effects, and restoring teeth with minimally invasive techniques. PMID- 15833011 TI - Refining occlusion with muscle balance to enhance long-term orthodontic stability. AB - The primary objective of orthodontic treatment is the movement of teeth into a more ideal esthetic and functional relationship. Finishing a case with the muscles of mastication in equilibrium is another very important objective that often does not receive enough consideration. If the occlusal forces in maximum intercuspation are distributed unevenly around the arch, tooth movement most likely will occur and an endless procession of retainers will be necessary for retention. Today, it is possible to make simultaneous and precise measurements of the relative force of each occlusal contact, the timing of the occlusal contacts, and the specific muscle contraction levels. This technological breakthrough represents a paradigm shift in thinking and may improve orthodontic stability. PMID- 15833012 TI - Use of an erbium laser in lieu of retraction cord: a modern technique. AB - The two-cord retraction technique is used for troughing around a crown to achieve biologic width. This technique can cause significant discomfort for the patient and offers several potential clinical disadvantages for the dentist, of which unpredictable tissue recession is the most significant. The clinical case presented in this article compares the use of the standard two-cord retraction technique with that of an 2,780 nm erbium-class dental laser to determine which method achieves an accurate, easily readable impression while respecting the biologic width. Using an erbium laser to achieve the trough prior to placing an indirect restoration results in little or no postoperative discomfort for the patient; in addition, the erbium laser reduces intraoperative complications related to tissue recession and patient discomfort while providing consistently accurate impressions. PMID- 15833013 TI - Clinical case report: capillary hemangioma. AB - This article reviews a case involving a patient with multiple hopeless dentition, massive gingival hyperplasia, and a vascular lesion. The hopeless dentition was extracted, exophytic hyperplastic tissue was removed, and the lesion was involuted following surgical intervention. The patient's occlusion subsequently was restored with a partial denture. Patients with multiple pathologies require careful evaluation to determine the level of care and sequence of treatment that is required. PMID- 15833014 TI - Subcutaneous cervicofacial emphysema and pneumo-mediastinum: a rare complication after a crown preparation. AB - Subcutaneous cervicofacial emphysema and pneumo-mediastinum are relatively rare complications from different dental procedures (including oral surgery and restorative treatment). Many cases go unrecognized or are misdiagnosed. While the majority of cases resolve spontaneously, some can lead to life-threatening complications that require emergency intervention. This article presents a case report of subcutaneous and mediastinal emphysema following a routine restorative dental procedure. The differential diagnosis and management of this condition is discussed. PMID- 15833015 TI - Dental treatment and orthopedic implants--a malpractice case. AB - A patient who required a revision of a six-year-old hip prosthesis five months following root canal therapy and removal of a root tip subsequently sued his dentist for malpractice. The alleged torts were the presence of a positive culture for lactobacillus--presumably an "oral organism" from the infected hip site one week following the dental procedures--and improper use of antibiotics by the dentist. This case underscores the need for the medical and dental communities to agree on what constitutes proper treatment for dental infections. In addition, the arbitrary use of antibiotics, which has led to increasing numbers of bacterial-resistant organisms and other side effects, needs to be addressed. The guidelines for prophylactic antibiotic administration prior to dental treatment in orthopedic implant patients must be revised significantly, based upon risk-to-benefit ratio, documented research, and the literature. PMID- 15833016 TI - The patient with a transient bacteremia. AB - A significant percentage of antibacterial agents that dentists prescribe are used for the prevention of, rather than the treatment of, an established infection. In general, when a single effective drug is used to prevent infection from a specific microorganism--or to eradicate it either immediately or soon after it has become established--chemoprophylaxis frequently is successful. Consequently, chemoprophylaxis may be appropriate for preventing secondary bacterial infection in patients who are ill with other diseases. In situations where bacteremia is highly predictable, it would seem wise to administer prophylactic antibiotics for high-risk patients to minimize morbidity and mortality; however, clinicians should avoid "reflex prescribing" or "rational activism." Before instituting antibacterial chemotherapy to prevent illness, dentists should weigh the benefits and risks to the patient as well as to the community. PMID- 15833017 TI - Digital dental radiography needs assessment utilizing the Delphi process. AB - This article reports the results of a needs assessment using the Delphi process. The purpose of this study was to determine the architecture and equipment (radiographic and computer network) for establishing and supporting an enterprise's vast digital dental radiographic program. Two panels of subject matter experts identified the needs for a digital dental radiography system; one expert panel consisted of dentists while the other panel's members were information management/information technology (IM/IT) experts. Panelists rated digital dental radiography needs using descriptive statistics and a 7-point bipolar scale. Among dentists, the four most important needs were the capability to store images, image appearance, centralized image storage, and a dedicated back-up server; among IM/IT panelists, they were ensuring HIPAA compliance, use of a standard formatting procedure, excellent print quality, and use of encryption for data transmission. This study's findings are useful for any organization seeking to initiate or improve its own digital radiography program. PMID- 15833018 TI - The use of systemic antibiotics in the treatment of aggressive periodontal disease. AB - General dentists frequently encounter patients with aggressive periodontal disease and should be able to diagnose and manage this disease properly. Periodontal care in the absence of a comprehensive treatment plan and proper therapy can result in the rapid progression of the disease and, ultimately, tooth loss. It is important for the general dentist to diagnose, inform, and treat the periodontal patient accurately, using referral and nonsurgical, surgical, and antimicrobial/antibiotic therapy. This article provides a brief history of the classification of aggressive periodontal disease, describes the microorganisms associated with aggressive periodontal disease, discusses the selection and use of systemic antibiotics in therapy, and lists the various antibiotic regimens for treating aggressive periodontal disease. PMID- 15833019 TI - Nonhealing extraction site. PMID- 15833020 TI - Yellowish nodule of the mandibular gingiva. PMID- 15833021 TI - Characterization of an acute molecular marker of nongenotoxic rodent hepatocarcinogenesis by gene expression profiling in a long term clofibric acid study. AB - Evaluation of the nongenotoxic potential early during the development of a drug presents a major challenge. Recently, two genes were identified as potential molecular markers of rodent hepatic carcinogenesis: transforming growth factor beta stimulated clone 22 (TSC-22) and NAD(P)H cytochrome P450 oxidoreductase (CYP R) (1). They were identified after comparing the gene expression profiles obtained from the livers of Sprague-Dawley rats treated with different genotoxic and nongenotoxic compounds in a 5 day repeat dose in vivo study. To assess the potential of these two genes as acute markers of carcinogenesis, we investigated their modulation during a long-term nongenotoxic study in the rat using a classic initiation-promotion regime. Clofibric acid (CLO), which belongs to the broad class of chemicals known as peroxisome proliferators, was used as a nongenotoxic hepatocarcinogen. Male F344 rats were given a single nonnecrogenic injection of diethylnitrosamine (0 or 30 mg/kg) and fed a diet containing none or 5000 ppm CLO for up to 20 months. Necropsies of five rats per groups were performed at 18, 46, 102, 264, 377, 447 (control, DEN, and DEN + CLO rats), 524, and 608 days (for the CLO and control rats). Gross macroscopic and microscopic evaluation and gene expression profiling (on Affymetrix microarrays) were performed in peritumoral and tumoral liver tissues. Bioanalysis of the liver gene expression data revealed that TSC-22 was strongly down-regulated early in the study. Its underexpression was maintained throughout the study but disappeared upon CLO withdrawal. These modulations were confirmed by real-time polymerase chain reaction. However, CYP-R gene expression was not significantly altered in our study. Taken together, our results showed that TSC-22, but not CYP-R, has the potential to be an acute early molecular marker for nongenotoxic hepatocarcinogenesis in rodents. PMID- 15833022 TI - Phenotypic anchoring of global gene expression profiles induced by N-hydroxy-4 acetylaminobiphenyl and benzo[a]pyrene diol epoxide reveals correlations between expression profiles and mechanism of toxicity. AB - The goal of this study was to compare changes in gene expression induced by exposure to different carcinogens and to anchor these changes to the induced levels of toxicity and mutagenesis. The human TK6 lymphoblastoid cell line was used as an in vitro model system, and reactive metabolites of two human carcinogens, benzo[a]pyrene and 4-aminobiphenyl, were used as model compounds. We first determined the toxicity of the model compounds N-hydroxy-4 acetylaminobiphenyl (N-OH-AABP) and benzo[a]pyrene diol epoxide (BPDE) in TK6 cells. BPDE was about 1000-fold more toxic and mutagenic than N-OH-AABP in TK6 cells on a molar basis. We next treated cells with three doses of each compound that resulted in low, medium, and high toxicities (5, 15, and 40%) and harvested cells at different times after exposure. Using comparable levels of toxicity as the phenotypic anchor, we compared the patterns of gene expression induced by each reactive metabolite using printed cDNA microarrays comprising approximately 18,000 human gene/EST sequences. The microarray data from the N-OH-AABP and BPDE treatment groups were compared using self-organizing map clustering algorithms, as well as a statistical regression modeling approach. While subsets of genes indicative of a generalized stress response [Hsp 40 homologue (DNAJ), Hsp70, Hsp105, and Hsp 125] were detected after exposure to both compounds at all concentrations, there were also many differentially regulated genes, including phase I xenobiotic metabolism [e.g., glutathione transferase omega (GSTTLp28) and antioxidant enzymes (Apxl)]. Other differentially regulated genes included those encoding proteins involved in all major DNA repair pathways, including excision repair (e.g., ERCC5), mismatch repair (e.g., MLH3), damage specific DNA binding protein (e.g., DDB2), and cisplatin resistance-associated overexpressed protein (LUC7A, CRA). Differences in the transcriptional response of TK6 cells to N-OH AABP or BPDE exposure may explain the dramatic differences in the toxicity and mutagenicity of these human carcinogens. PMID- 15833023 TI - Negative ion tandem mass spectrometry for the detection of glutathione conjugates. AB - Characterization of S-linked conjugates of the endogenous tripeptide glutathione (gamma-glutamyl-cysteinylglycine, GSH) represents a valuable indirect approach for the identification of chemically reactive, electrophilic intermediates formed during the metabolism of both foreign compounds and endogenous substances. In most cases, GSH adducts generated in vitro or excreted in the bile of animals are detected by the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS), employing survey scans based on characteristic fragmentations of this class of conjugates. However, a limitation of current LC-MS/MS approaches, which typically employ electrospray ionization with analysis of positive ions, is that no single survey scan exhibits broad utility in the detection of unknown GSH adducts, since different structural classes of conjugate (aromatic, benzylic, aliphatic, thioester, etc.) behave differently upon collision-induced dissociation (CID) of the respective [M + H]+ parent ions. In the present study, we evaluated MS/MS in the negative ion mode as an alternative approach and report herein that the spectra obtained by CID of the [M - H]- ions of a number of representative GSH adducts, as well as GSH itself, are dominated by fragments originating from the glutathionyl moiety of the tripeptide. In particular, the anion at m/z 272, corresponding nominally to deprotonated gamma-glutamyl-dehydroalanyl-glycine, was abundant in the negative ion spectra of free GSH and all GSH conjugates examined, suggesting that scanning for precursors of this ion may provide a generally applicable technique for the detection of adducts of unknown structure. The utility of this novel detection strategy was demonstrated in a series of in vitro and in vivo experiments where compounds known to undergo metabolic activation were examined for their propensity to form conjugates with GSH. In all cases, scanning for precursors of m/z 272 in the negative ion mode revealed the presence of the expected adducts and in some instances revealed additional conjugates that had not been reported previously. Positive ion MS/MS, on the other hand, was more useful than the corresponding negative ion scans in providing information on the molecular structure of GSH conjugates. PMID- 15833024 TI - Metabonomic deconvolution of embedded toxicity: application to thioacetamide hepato- and nephrotoxicity. AB - We present here the potential of an integrated metabonomic strategy to deconvolute the biofluid metabolic signatures in experimental animals following multiple organ toxicities, using the well-known hepato- and nephrotoxin, thioacetamide. Male Han-Wistar rats were dosed with thioacetamide (150 mg/kg, n = 25), and urine, plasma, liver, and kidney samples were collected postdose for conventional NMR and magic angle spinning (MAS) NMR spectroscopy. These data were correlated with histopathology and plasma clinical chemistry collected at all time points. 1H MAS NMR data from liver and kidney were related to sequential 1H NMR measurements in urine and plasma using pattern recognition methods. One dimensional 1H NMR spectra were data-reduced and analyzed using principal components analysis (PCA) to show the time-dependent biochemical variations induced by thioacetamide toxicity. From the eigenvector loadings of the PCA, those regions of the 1H NMR spectra, and hence the combinations of endogenous metabolites marking the main phase of the toxic episode, were identified. The thioacetamide-induced biochemical manifestations included a renal and hepatic lipidosis accompanied by hypolipidaemia; increased urinary excretion of taurine and creatine concomitant with elevated creatine in liver, kidney, and plasma; a shift in energy metabolism characterized by depleted liver glucose and glycogen; reduced urinary excretion of tricarboxylic acid cycle intermediates and raised plasma ketone bodies; increased levels of tissue and plasma amino acids leading to amino aciduria verifying necrosis-enhanced protein degradation and renal dysfunction; and elevated hepatic and urinary bile acids indicating secondary damage to the biliary system. This integrated metabonomic approach has been able to identify the tissue of origin for biomarkers present in the metabolic profiles of biofluids, following the onset and progression of a multiorgan pathology, and as such highlights its potential in the evaluation of embedded toxicity in novel drug candidates. PMID- 15833025 TI - Differential removal of DNA adducts derived from anti-diol epoxides of dibenzo[a,l]pyrene and benzo[a]pyrene in human cells. AB - The polycyclic aromatic hydrocarbons (PAHs) dibenzo[a,l]pyrene (DBP) and benzo[a]pyrene (BP) are widespread environmental contaminants and potent carcinogens. The fjord-region DBP is considerably more carcinogenic than the bay region BP. This fact can be ascribed to differences in DNA binding efficiency of their ultimate carcinogenic diol epoxide (DE) intermediates, differences in structural features of the DNA adducts, and differences in DNA adduct recognition and the subsequent lesion removal by nucleotide excision repair (NER). We have compared the formation and removal of adducts as a function of time formed by the carcinogenic metabolites (-)-anti-DBPDE and (+)-anti-BPDE in A549 human epithelial lung carcinoma cells. Cells were exposed to 0.1 or 1.0 microM (-)-anti DBPDE and (+)-anti-BPDE, respectively. Adducts were measured at various post treatment times (up to 6 h) by enzymatic DNA hydrolysis and a HPLC procedure that allows monitoring of all cis- and trans-nucleoside adducts of dA and dG. Treatment with 0.1 microM (-)-anti-DBPDE resulted in an initial increase of adducts to a maximal level of 144 pmol adducts/mg of DNA after 1 h of incubation. This was followed by an apparent, although not statistically significant, slow removal of adducts. After 6 h of incubation, at least 80% seems to remain. In cells treated with 1.0 microM (+)-anti-BPDE, the maximal level of 140 pmol adducts/mg of DNA was reached within 20 min of exposure. The formation was followed by an initial rapid decline in the adduct level (1.54 pmol adducts/mg of DNA/min) and a later statistically significant slower rate (0.14 pmol adducts/mg of DNA/min) of adduct removal. After 1 h of incubation, about 45% of the adducts are removed followed by 75% at 6 h. The biphasic pattern of BPDE removal has been observed previously in mammalian cells and, at least in part, may reflect the action of transcription-coupled repair (TCR) and the subsequent global genomic repair (GGR). Comparing the rate of removal of adducts derived from BPDE with those of DBPDE, the latter are obviously more refractory to the NER-coupled repair than the former. Furthermore, the apparent resistance of adducts from DBPDE to be eliminated may reflect the ability of such adducts to escape recognition and/or the subsequent removal by the NER machinery. Further analysis of DNA adduct distribution as a function of incubation time reveals that the dA/dG adduct ratio for BPDE was independent of time (4% dA, 96% dG), whereas the corresponding ratio for DBPDE was significantly increased from 2.9 (74% dA, 26% dG) at 20 min to 4.0 (80% dA, 20% dG) after 6 h of incubation. The results presented here on DNA adduct removal in mammalian cells are in part consistent with recent results on NER-coupled activity on bay- and fjord-region DE-modified oligonucleotides in vitro and further substantiate the hypothesis that the high carcinogenicity of the nonplanar PAHs arise from the ability of the preferentially formed dA adducts to escape recognition by surveillance systems and the subsequent NER-coupled lesion removal. PMID- 15833026 TI - On the generation and outcome of 3-(N-phenylamino)propane-1,2-diol derivatives in deodorized model oils related to toxic oil syndrome. AB - Toxic Oil Syndrome (TOS) was a massive food-born intoxication that occurred in Spain in 1981 and affected more than 20,000 people. TOS was attributed to the ingestion of rapeseed oil that had been adulterated with aniline, illegally refined, and delivered for human consumption. Two chemical species derived from aniline have been identified in oil batches: fatty acid anilides, qualified as biomarkers of the adulterated oil, and fatty acid esters of 3-(N phenylamino)propane-1,2-diol (PAP), considered toxic oil biomarkers. These esters were generated by chemical processes during oil refining, specifically in the deodorization step, which involves treatment of the oil at high temperatures under vacuum to remove volatile contaminants. Since PAP derivatives are strongly associated with TOS, their formation and putative interconversion in a toxic oil model has been studied. The main results obtained are (i) only triglycerides and aniline are required to produce PAP esters, thus eliminating the possibility that unknown activators present in the deodorization tank were required for toxification of the oil; (ii) PAP and PAP mono- and diesters are chemically interrelated, as are anilides and PAP esters to an even higher degree. In addition to the reaction of aniline with triglycerides, anilides can be also formed via attack of PAP esters by aniline. However, the most important source of anilides during deodorization seems to be the thermal decomposition of PAP esters. Overall, these results suggest that the generation and outcome of PAP derivatives during deodorization is a complex scenario whereby PAP esters are not only generated from different reactions but decompose to produce anilides, among other compounds. In addition to providing a rapeseed oil model that reproduces the composition of case oils with respect to anilides and PAP derivatives, the results presented herein further support the hypothesis imputing PAP diesters or their metabolites for the intoxication episode. PMID- 15833027 TI - In vitro bioactivation of dihydrobenzoxathiin selective estrogen receptor modulators by cytochrome P450 3A4 in human liver microsomes: formation of reactive iminium and quinone type metabolites. AB - Estrogens and selective estrogen receptor modulators (SERMs) are prescribed widely in the clinic to alleviate symptoms in postmenopausal women, and they are metabolized to reactive intermediates, which may elicit adverse effects. As part of our efforts to develop safer SERMs, in vitro covalent protein binding of (2S,3R)-(+)-3-(4-hydroxyphenyl)-2-[4-(2-piperidin-1-ylethoxy)phenyl]-2,3-dihydro 1,4-benzoxathiin-6-ol (I) was evaluated. Radioactivity from [3H]I became covalently bound to proteins in a fashion that was both time- and NADPH-dependent in human liver microsomes and reached a value of 1106 pmol equiv/mg protein following a 45 min incubation. At least three pathways are involved in the bioactivation of I, namely, oxidative cleavage of the dihydrobenzoxathiin moiety to give a hydroquinone/para-benzoquinone redox couple, hydroxylation at position 5 or 7 of the benzoxathiin moiety leading to an o-quinone intermediate, and metabolism of the piperidine ring to give an iminium ion. The latter reactive intermediate was identified as its bis-cyano adduct when human liver microsomal incubations were performed in the presence of sodium cyanide. Structural modification of I, including a replacement of the piperidine with a pyrrolidine group, led to (2S,3R)-(+)-3-(3-hydroxyphenyl)-2-[4-(2-pyrrolidin-1 ylethoxy)phenyl]-2,3-dihydro-1,4-benzoxathiin-6-ol (II), which did not form a reactive iminium ion. Following the incubation of II with human liver microsomes, covalent binding to proteins was reduced (461 pmol equiv/mg protein), the residual level of binding apparently due to the formation of a rearranged biphenyl quinone type metabolite. Studies with inhibitory antibodies and chemical inhibitors showed that P450 3A4 was the primary enzyme responsible for oxidative bioactivation of I and II in human liver microsomes. These studies thus demonstrated that gaining an understanding of bioactivation mechanisms may be exploited in terms of guiding structural modifications of drug candidates to minimize covalent protein binding and, hopefully, to lower the potential for drug mediated adverse effects. PMID- 15833028 TI - Formation of benzylic-DNA adducts resulting from 7,12-dimethylbenz[a]anthracene in vivo. AB - Studies were undertaken to determine the formation of benzylic-DNA adducts in rats administered 7,12-dimethylbenz[a]anthracene (DMBA) and its meso-region metabolites by subcutaneous injection. Here, we show that 7-hydroxymethyl-12 methylbenz[a]anthracene (7-HMBA) and 7-sulfoxymethyl-12-methylbenz[a]anthracene (7-SMBA) gave rise to some benzylic-DNA adducts indistinguishable from adducts formed from DMBA. Adducts were analyzed by butanol enrichment-mediated 32P postlabeling assay. Female Sprague-Dawley rats given a combined dose of 420 micromol DMBA/kg b. wt resulted in two major and up to nine minor adducts in the subcutaneous tissue, with chromatographic resemblance to benzylic-DNA adducts prepared in vitro. Subcutaneous administration of 7-HMBA, 7-SMBA, and 7-methyl-12 hydroxymethylbenz[a]anthracene (12-HMBA) (210, 42, and 210 micromol/kg b. wt, respectively) each resulted in one major and several minor benzylic-DNA adducts. From cochromatography with reference adducts, it was concluded that the benzylic DNA adduct 4, derived from the parent compound, comigrates with the major adduct from 7-HMBA and 7-SMBA, whereas adducts 2 and 3 comigrate with adducts resulting from 12-HMBA and 7-methyl-12-sulfooxymethylbenz[a]anthracene, respectively. These data suggest that 7-sulfooxymethyl- and 12-sulfooxymethy derivatives produce distinct adducts. Several major and minor diol epoxide-related DNA adducts were also detected. The diol epoxide- and benzylic-DNA adducts were found in a 2:1 ratio. The oral, intraperitoneal, and intramammiliary treatments with DMBA showed no detectable benzylic adducts in the liver and mammary glands 24 h after the last treatment, although the adduct formation was clearly evident with SMBA and/or HMBA treatments, suggesting that hydroxylation of DMBA to form HMBA may be the rate-limiting step for the meso-methyl substitution pathway. The present study clearly demonstrates the in vivo formation of benzylic-DNA adducts from DMBA. The data also reveal the involvement of the 12-methyl group of DMBA in adduct formation. PMID- 15833029 TI - Detection and quantification of N-(deoxyguanosin-8-yl)-4-aminobiphenyl adducts in human pancreas tissue using capillary liquid chromatography-microelectrospray mass spectrometry. AB - Cigarette smoking has been associated with various cancers including bladder and pancreas. 4-Aminobiphenyl has been isolated as a constituent of cigarette smoke and has been established as a carcinogen in various animal models and humans. In rodents and humans, 4-aminobiphenyl is N-hydroxylated and forms adducts to DNA, the predominant one being N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8-ABP). In this study, we report a micro-electrospray mass spectrometric (muESI-MS) isotope dilution method for the detection and quantification of dG-C8-ABP in human pancreatic tissue. A reverse phase capillary column (320 microm ID) was connected to a triple quadrupole mass spectrometer via a commercially available micro-ESI source. The system was operated in the selected reaction monitoring mode transmitting the [M + H]+ --> [M + H - 116]+ transitions for both the analyte and the isotopically labeled internal standard. Twelve human pancreas samples were analyzed, where six were current smokers (three male and three female) and six were considered nonsmokers (three female and three male). Of the samples analyzed, six showed dG-C8-ABP levels above the limit of quantification for the method, five were considered to have levels that were undetectable, and one was discarded due to inconsistent internal standard signal. The age of the human subjects ranged from 17 to 63, and, in samples where adduct was present, levels ranged anywhere from 1 to 60/10(8) nucleotides. Although no correlation between smoking preference, age, or gender was proven with this particular sample pool, this report demonstrates that capillary LC-muESI-MS can provide a sensitive and definitive method for DNA adduct analysis in human tissue. PMID- 15833030 TI - Microsomal biotransformation of benzo[ghi]perylene, a mutagenic polycyclic aromatic hydrocarbon without a "classic" bay region. AB - Carcinogenic polycyclic aromatic hydrocarbons (PAH), e.g., benzo[a]pyrene (BaP), possess a bay region comprising an ortho-fused benzene ring. Benzo[ghi]perylene (BghiP) represents the group of PAHs lacking such a "classic" bay region and hence cannot be metabolically converted like BaP to bay region dihydrodiol epoxides considered as ultimate mutagenic and carcinogenic metabolites of PAH. BghiP exhibits bacterial mutagenicity in strains TA98 (1.3 his(+)-revertant colonies/nmol) and TA100 (4.3 his(+)-revertant colonies/nmol) of Salmonella typhimurium after metabolic activation by the postmitochondrial hepatic fraction of CD rats treated with 3-methylcholanthrene. Inhibition of microsomal epoxide hydrolase (mEH) with 1,1,1-trichloro-2-propene oxide raised the bacterial mutagenicity of BghiP in TA98 almost 4-fold indicating arene oxides as ultimate mutagens. To confirm this assumption, the biotransformation of BghiP was elucidated. Incubation of BghiP with liver microsomes of CD rats treated with Aroclor 1254 yielded 17 ethyl acetate extractable metabolic products. Twelve metabolites were identified by a combination of chromatographic, spectroscopic, and biochemical methods. The microsomal biotransformation of BghiP proceeds by two pathways: Pathway I starts with the monooxygenase attack at the 7-position leading to the 7-phenol, which is transformed to the 7,8- and 7,10-diphenols followed by oxidation to the 7,8- and 7,10-quinones. On pathway II, the K regions of BghiP are successively converted to arene oxides yielding the indirectly identified 3,4-oxide and the 3,4,11,12-bisoxides. Enzymatic hydrolysis of the 3,4 oxide leads to the trans-3,4-dihydrodiol, which is oxidized to the 3,4-quinone. Similarly, the trans-3,4-trans-11,12-bisdihydrodiols and the trans-3,4 dihydrodiol 11,12-quinone are generated from the 3,4,11,12-bisoxides. The trans 3,4-dihydrodiol and the trans-3,4-trans-11,12-bisdihydrodiols are preferentially formed as R,R and R,R,R,R enantiomers, respectively. The intrinsic bacterial mutagenicity of the 3,4,11,12-bisoxides is rather low and hardly explains the strong increase in bacterial mutagenicity of BghiP after inhibition of mEH. Thus, we believe that the 3,4-oxide plays a more important role as the ultimate mutagenic metabolite of BghiP. PMID- 15833031 TI - Synthesis and properties of an acetaldehyde-derived oligonucleotide interstrand cross-link. AB - Acetaldehyde (AA), occurring widely in the human environment, is a mutagen and carcinogen. AA can react with DNA to form AA-DNA adducts. Several types of adducts, including an interstrand cross-link 3-(2-deoxyribos-1-yl)-5,6,7,8 tetrahydro-8-(N2-deoxyguanosyl)-6-methylpyrimido[1,2-a]purine-10(3H)one (7), have been previously characterized by our laboratory. We hypothesize that cross-link 7 may be involved in determining the mutagenic and carcinogenic properties of AA. To address this question, the double-stranded oligonucleotide 13, bearing cross link 7, was synthesized in a sequence appropriate for mutagenicity studies in human cells. Oligonucleotide 9, containing 2-fluoro-O6 (trimethylsilylethyl)deoxyinosine (dIno), was reacted with 4-amino-1,2 pentanediol, followed by treatment with NaIO4. The resulting oligonucleotide 11 containing the 1,N2-propano-deoxyguanosine (dGuo) 5 was incubated with the complementary oligonucleotide 12 to give the desired cross-link 13, which was characterized by negative-ion electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) and enzymatic hydrolysis to cross-link 7. The formation of cross-link 13 at 5'-CpG-3' was confirmed by incubation of 11 with [15N5]12 containing a 5'-Cp[15N5]G-3' sequence. The formation of cross-link 13 was reversible. Therefore, oligonucleotide 24 containing the irreversible analogue of cross-link 7, 1,3-bis(2'-deoxyguanos-N2-yl)butane, was synthesized for use as a control in the mutagenicity studies. Oligonucleotide 21 was reacted with 1,3-diaminobutane dihydrochloride, followed by incubation with the complementary oligonucleotide 23, to give 24. To determine the optimum distance and orientation for cross-link formation, six oligonucleotides, containing 5 at the i + 1, i + 2, and i + 3 or the i - 1, i - 2, and i - 3 positions relative to dGuo in the complementary strand, were 5'-end labeled with [gamma-32P]ATP, followed by incubation of each labeled oligonulceotide with its complementary strand and then analysis by denaturing polyacrylamide gel electrophoresis. Only the oligonucleotide containing 5'-Cp5-3' formed the cross-link with the complementary 5'-CpG-3' sequence. The results of this study confirm the structure of an AA-derived DNA cross-link, supply characterized cross-link-containing oligonucleotides for mutagenicity studies, and provide information on the optimum distance and orientation for cross-link formation. PMID- 15833032 TI - The acid-promoted reaction of the green tea polyphenol epigallocatechin gallate with nitrite ions. AB - Exposure of 400 microM (-)-epigallocatechin-3-O-gallate (EGCG), the main polyphenolic constituent of green tea, to equimolar concentrations of nitrite ions in 0.5 M HCl at 37 degrees C resulted in the formation of a distinct pattern of products that were trapped as phenazine derivatives by treatment with o phenylenediamine. Repeated chromatographic fractionation eventually allowed isolation of four main species, which were identified by 2D NMR and MS analysis as 1b, derived from EGCG quinone 1a, the isomeric oximes 2b,c, arising from nitrosation of EGCG on the pyrogallol B-ring, and the dioxime 4b in which the A ring was doubly substituted. At lower EGCG concentrations (e.g., 25 microM) and at pH 3, reaction with equimolar amounts of nitrite gave 1b as the first formed species, whereas nitrosation products 2b,c and 4b became detectable only with excess nitrite. Similar reaction of chemically prepared 1a with acidic nitrite led to the formation of 2b,c and 4b, suggesting that this quinone may be an intermediate in the nitrosation of EGCG. Exposure of green tea extracts to acidic nitrite ions resulted in the conversion of EGCG to 1a, detected as 1b. Overall these results substantiate literature reports suggesting that the protective effects of EGCG against nitrosation involve mainly an initial redox exchange process and hint at a hitherto unrecognized property of quinone 1a as a potential scavenger of nitrosating species. PMID- 15833033 TI - Reaction of glyoxal with 2'-deoxyguanosine, 2'-deoxyadenosine, 2'-deoxycytidine, cytidine, thymidine, and calf thymus DNA: identification of DNA adducts. AB - Glyoxal (ethanedial) is an increasingly used industrial chemical that has been found to be mutagenic in bacteria and mammalian cells. In this study, the reactions of glyoxal with 2'-deoxyguanosine, 2'-deoxyadenosine, 2'-deoxycytidine, cytidine, thymidine, and calf thymus DNA have been studied in aqueous buffered solutions. The nucleoside adducts were isolated by reversed-phase liquid chromatography and characterized by their UV absorbance and 1H and 13C NMR spectroscopic and mass spectrometric features. The reaction with 2' deoxyguanosine gave one adduct, the previously known 3-(2'-deoxy-beta-D-erythro pentofuranosyl)-5,6,7-trihydro-6,7-dihydroxyimidazo[1,2-a]purine-9-one adduct. The reaction of 2'-deoxyadenosine with glyoxal resulted in the formation of a previously not reported N6-(hydroxyacetyl)-2'-deoxyadenosine adduct. In the reaction of glyoxal with 2'-deoxycytidine and cytidine at neutral conditions and 37 degrees C, 5-hydroxyacetyl pyrimidine derivatives were obtained. When the cytidine reaction was performed at pH 4.5 and 50 degrees C, the 5-hydroxyacetyl derivative of uridine was formed through deamination of cytidine-glyoxal. Adducts in the thymidine reaction could not be detected. In the reaction of glyoxal with calf thymus DNA, the 2'-deoxyguanosine-glyoxal and 2'-deoxyadenosine-glyoxal adducts were obtained, the former being the major adduct. PMID- 15833034 TI - An automated group contribution method in predicting aquatic toxicity: the diatomic fragment approach. AB - We developed a group contribution method (GCM) to correlate acute toxicity (96 h LC50) for the fathead minnow (Pimephales promelas) for 607 organic chemicals. Unlike most of the existing methods, the new one makes no use of predefined groups as descriptors. A simple general rule is proposed to break down any molecule into diatomic fragments. The entire data set was partitioned three times. Each time, a training set and a test set were obtained with a ratio of 2:1. For each partition quantitative structure-activity relationship, models were developed using Powell's minimization method, multilinear regression, neural networks, and partial least squares. The GCM method achieved a good correlation of the data for both training and test sets, regardless of the partition considered. The method is therefore robust and can be generally applied. Further model improvements are described. PMID- 15833035 TI - Reaction kinetics of biotinylated organophosphorus toxicant, FP-biotin, with human acetylcholinesterase and human butyrylcholinesterase. AB - A biotinylated organophosphate could be useful for identifying proteins that react with organophosphorus toxicants (OP). FP-biotin, 10 (fluoroethoxyphosphinyl)-N-(biotinamidopentyl)decanamide, was synthesized and found to be stable in methanol and chloroform but less stable in water. Because acetylcholinesterase (AChE, EC 3.1.1.7) and butyrylcholinesterase (BChE, EC 3.1.1.8) are known to be sensitive targets of OP, their reactivity with FP-biotin was tested. The rate constant for reaction with human AChE was 1.8 x 10(7) M(-1) min(-1), and for human BChE, it was 1.6 x 10(8) M(-1) min(-1). A phosphorus stereoisomer, constituting about 50% of the FP-biotin preparation, appeared to be the reactive species. The binding affinity was estimated to be >85 nM for AChE and >5.8 nM for BChE. It was concluded that FP-biotin is a potent OP, well-suited for searching for new biomarkers of OP exposure. PMID- 15833036 TI - Aminoacetone induces oxidative modification to human plasma ceruloplasmin. AB - Aminoacetone (AA), a putative endogenous source of cytotoxic methylglyoxal, and ceruloplasmin (CP), the antioxidant plasma copper transporter, are known to increase in diabetes. AA was recently shown in vitro to act as a pro-oxidant toward ferritin and isolated mitochondria. We now report AA oxidative effects on CP mediated by AA-generated reactive oxygen species (ROS). Incubation of 1.5 microM human CP with 0.05-1 mM AA resulted in extensive protein aggregation. That ROS-driven thiol cross-linking underlies the CP aggregation was evidenced by the inhibitory effects of added superoxide dismutase, catalase, mannitol, and dithiothreitol. The addition of CP to AA (mM) solutions accelerated oxygen consumption by AA and caused CP copper ion release and loss of ferroxidase and aminoxidase activities. If operative in vivo, this reaction would impair the antioxidant role of CP and iron uptake by ferritin and hence contribute to intracellular iron-induced oxidative stress during AA accumulation in diabetes mellitus. PMID- 15833037 TI - Effects of benzene metabolites on DNA cleavage mediated by human topoisomerase II alpha: 1,4-hydroquinone is a topoisomerase II poison. AB - Although benzene induces leukemias in humans, the compound is not believed to generate chromosomal damage directly. Rather, benzene is thought to act through a series of phenolic- and quinone-based metabolites, especially 1,4-benzoquinone. A recent study found that 1,4-benzoquinone is a potent topoisomerase II poison in vitro and in cultured human cells [Lindsey et al. (2004) Biochemistry 43, 7363 7374]. Because benzene is metabolized to multiple compounds in addition to 1,4 benzoquinone, we determined the effects of several phenolic metabolites, including catechol, 1,2,4-benzenetriol, 1,4-hydroquinone, 2,2'-biphenol, and 4,4' biphenol, on the DNA cleavage activity of human topoisomerase II alpha. Only 1,4 hydroquinone generated substantial levels of topoisomerase II-mediated DNA scission. DNA cleavage with this compound approached levels observed with 1,4 benzoquinone (approximately 5- vs 8-fold) but required a considerably higher concentration (approximately 250 vs 25 microM). 1,4-Hydroquinone is a precursor to 1,4-benzoquinone in the body and can be activated to the quinone by redox cycling. It is not known whether the effects of 1,4-hydroquinone on human topoisomerase II alpha reflect a lower reactivity of the hydroquinone or a low level of activation to the quinone. The high concentration of 1,4-hydroquinone required to increase enzyme-mediated DNA cleavage is consistent with either explanation. 1,4-Hydroquinone displayed attributes against topoisomerase II alpha, including DNA cleavage specificity, that were similar to those of 1,4 benzoquinone. However, 1,4-hydroquinone consistently inhibited DNA ligation to a greater extent than 1,4-benzoquinone. This latter result implies that the hydroquinone may display (at least in part) independent activity against topoisomerase II alpha. The present findings are consistent with the hypothesis that topoisomerase II alpha plays a role in the initiation of specific types of leukemia that are induced by benzene and its metabolites. PMID- 15833038 TI - Role of phenoxyl radicals in DNA adduction by chlorophenol xenobiotics following peroxidase activation. AB - Chlorophenol (CP) toxins are classified as probable human carcinogens and are known to undergo bioactivation to generate benzoquinone (BQ) electrophiles that react covalently with biopolymers. Recently, we characterized the ability of pentachlorophenol (PCP) to react covalently with deoxyguanosine (dG) following treatment with horseradish peroxidase (HRP)/H2O2 or myeloperoxidase to yield a C8 dG oxygen (O)-adduct that suggested the intermediacy of the pentachlorophenoxyl radical in covalent bond formation [Dai, J., Wright, M. W., and Manderville, R. A. (2003) Chem. Res. Toxicol. 16, 817-821]. Investigations currently focus on a wider range of CP substrates (PCP, 2,4,6-trichlorophenol (2,4,6-TCP), 2,4,5-TCP, and 2,4-dichlorophenol (2,4-DCP)) to establish their reactivity toward dG and duplex DNA (calf thymus (CT)) following activation by HRP/H2O2, as a representative peroxidase system. Our data show that chlorophenoxyl radicals may either react directly with dG and CT-DNA to form C8-dG O-adducts in an irreversible process or couple to yield 1,4-BQ electrophiles that react with dG to afford adducts of the benzetheno variety. These results are the first to establish the in vitro relevance of C8-dG O-adducts of phenolic toxins. The 1H NMR chemical shifts and reactivity of the benzetheno adducts favor 4' '-hydroxy 1,N2-benzetheno-dG adduct assignment, which is in contrast to other literature which has assigned the 1,4-BQ-dG adduct as 3' '-hydroxy-1,N2-benzetheno-dG. Overall, the results from this current study have provided new insights into peroxidase-mediated activation of CP substrates and have strengthened the hypothesis that direct reactions of phenoxyl radicals with DNA contribute to peroxidase-driven toxic effects of phenolic xenobiotics. PMID- 15833039 TI - 4-Hydroperoxy-2-nonenal-induced formation of 1,N2-etheno-2'-deoxyguanosine adducts. AB - Analysis of the reaction between 4-hydroperoxy-2-nonenal (HPNE) and 2' deoxyguanosine (dGuo) by liquid chromatography/mass spectrometry (LC/MS) revealed the formation of 1,N2-etheno-dGuo as well as heptanone-etheno-dGuo and trace amounts of dihydroxyheptane-etheno-dGuo. Identities of the dGuo adducts were confirmed by comparison with authentic standards. The minor dihydroxyheptane etheno-dGuo adducts could be generated from 2,3-epoxy-4-hydroxynonanal (EHN), the epoxidation product of 4-hydroxy-2-nonenal (HNE). An LC/MS method was developed for the analysis of EHN. No EHN was detected by LC/MS during the decomposition of HPNE. Therefore, the dihydroxyheptane-etheno-dGuo adducts are either generated from a direct reaction between HPNE and dGuo or from another intermediate that cannot be detected by LC/MS. In addition, no HNE-derived hydroxypropano-dGuo adducts were observed. On the basis of these findings, we conclude that HPNE, a primary product of lipid peroxidation, is a major precursor to the formation of 1,N2-etheno-dGuo. We propose that it arises from the reaction of dGuo and HPNE through the intermediate formation of a cyclic hydroxy-ethano-epoxide derivative. The minor amounts of heptanone-ethano-dGuo adducts that were formed from HPNE in the absence of vitamin C suggest that heptanone-etheno-dGuo can be generated directly from HPNE without the intermediate formation of ONE. Therefore, HPNE can be considered as another lipid hydroperoxide-derived bifunctional electrophile with the potential for biological activities that are similar to HNE and ONE. PMID- 15833043 TI - 10th annual meeting--Association for Molecular Pathology. PMID- 15833040 TI - A cancer roadmap. PMID- 15833044 TI - A 10-minute, US Food and Drug Administration-approved HIV test. AB - It is now an established fact that rapid HIV tests have important applications in a number of testing situations. Currently, four rapid HIV assays have been approved by the US Food and Drug Administration, including the one discussed in this review. A rapid, lateral flow HIV assay was evaluated at the University of Maryland, USA, one of several sites involved in the clinical evaluation. Samples used in the evaluation totaled 9000 and consisted of serum, plasma and venous whole-blood sets of samples from 3000 study subjects that included population groups considered to be at high risk for HIV infection (n=1000), from HIV positive individuals (n=1000), and from population groups considered at low risk for HIV infection (n=1000). US Food and Drug Administration-licensed screening and confirmatory assays were used as reference tests. The rapid assay exhibited a sensitivity of 100% across all three sample media and exhibited a specificity of 99.8% using whole blood and plasma and 99.7% using serum. This rapid assay is an excellent addition to the existing US Food and Drug Administration-approved rapid HIV tests, and provides versatility by allowing the testing of venipuncture and fingerstick whole-blood samples in addition to serum and plasma. PMID- 15833045 TI - Peptides in body fluids and tissues as markers of disease. AB - The general awareness of the importance of peptides in physiology and pathophysiology has increased strongly over the last few years. With worldwide progress in the analysis of whole genomes, the knowledge base in gene sequence and expression data useful for protein and peptide analysis has drastically increased. The medical need for relevant biomarkers is enormous. This is particularly true for the many types of cancer, but other diseases such as Type 2 diabetes also lack useful and adequate diagnostic markers with high specificity and sensitivity. Despite advances in imaging technologies for early detection of diseases, proteomic and peptidomic multiplex techniques have evolved in recent years. This review focuses on the application of peptidomics technologies to peptides in health and disease. Peptidomics technologies provide new opportunities for the detection of low-molecular-weight proteome biomarkers (peptides) by mass spectrometry. Improvements in peptidomics research are based on separation of peptides and/or proteins by their physicochemical properties in combination with mass spectrometric detection, identification and sophisticated bioinformatics tools for data analysis. Therefore, peptidomics technologies offer an opportunity to discover novel biomarkers for diagnosis and management of disease (e.g., prognosis, treatment decision and monitoring response to therapy). PMID- 15833046 TI - Applications of whole-genome high-density SNP genotyping. AB - The technology to simultaneously genotype hundreds of thousands of single nucleotide polymorphisms in a single assay has only recently been developed. These advances have the potential to revolutionize our ability to identify disease-associated proteins and their corresponding pathways as drugable targets. Several strategies that can take advantage of extremely high-density, genome-wide single nucleotide polymorphism genotyping to hone in on pathogenic genetic variants will be discussed. In familial linkage studies, high-density single nucleotide polymorphism genotyping has already been proven to speed up mutation identification of Mendelian traits several fold. Many studies now report examining loss of heterozygosity and genomic amplifications on a whole-genome level. Genotyping hundreds of thousands of single nucleotide polymorphisms in a single set of assays now also allows for whole-genome association studies in complex, multigenic diseases. The technology of high-density single nucleotide polymorphism genotyping has emerged rapidly, leaving data analysis and bioinformatic challenges only partially met. In this review, the immediate applications and implications of the rapidly changing high-density, whole-genome single nucleotide polymorphism genotyping field on translational research will be described. PMID- 15833047 TI - Tissue microarrays: applications in genomic research. AB - The widespread application of tissue microarrays in cancer research and the clinical pathology laboratory demonstrates a versatile and portable technology. The rapid integration of tissue microarrays into biomarker discovery and validation processes reflects the forward thinking of researchers who have pioneered the high-density tissue microarray. The precise arrangement of hundreds of archival clinical tissue samples into a composite tissue microarray block is now a proven method for the efficient and standardized analysis of molecular markers. With applications in cancer research, tissue microarrays are a valuable tool in validating candidate markers discovered in highly sensitive genome-wide microarray experiments. With applications in clinical pathology, tissue microarrays are used widely in immunohistochemistry quality control and quality assurance. The timeline of a biomarker implicated in prostate neoplasia, which was identified by complementary DNA expression profiling, validated by tissue microarrays and is now used as a prognostic immunohistochemistry marker, is reviewed. The tissue microarray format provides opportunities for digital imaging acquisition, image processing and database integration. Advances in digital imaging help to alleviate previous bottlenecks in the research pipeline, permit computer image scoring and convey telepathology opportunities for remote image analysis. The tissue microarray industry now includes public and private sectors with varying degrees of research utility and offers a range of potential tissue microarray applications in basic research, prognostic oncology and drug discovery. PMID- 15833048 TI - Molecular diagnostics by microelectronic microchips. AB - Molecular diagnostics is being revolutionized by the development of highly advanced technologies for DNA and RNA testing. One of the most important challenges is the integration of microelectronics to microchip-based nucleic acid technologies. The specific characteristics of these microsystems make the miniaturization and automation of any step of a molecular diagnostic procedure possible. This review describes the application of microelectronics to all the processes involved in a genetic test, particularly to sample preparation, DNA amplification and sequence variation detection. PMID- 15833049 TI - Improved diagnostic approaches to infection/sepsis detection. AB - Sepsis is a major healthcare problem from the perspective of mortality and economics. Advances in diagnostic detection of infection and sepsis have been slow, but recent advances in both soluble biomarker detection and quantitative cellular measurements promise the availability of improved diagnostic techniques. Though the promise of cytokine measurements reaching clinical practice have not matured, procalcitonin levels are currently available in many countries and appear to offer enhanced diagnostic distinction between bacterial and viral etiologies. Cellular diagnostics is poised to enter clinical laboratory practice in the form of neutrophil CD64 measurements, which offer superior sensitivity and specificity to conventional laboratory assessment of sepsis. Neutrophil CD64 expression is negligible in the healthy state. However, it increases as part of the systemic response to severe infection or sepsis. The combination of cellular proteomics, as in the case of neutrophil CD64 quantification, and selected soluble biomarkers of the inflammatory response, such as procalcitonin or triggering receptor expressed on myeloid cells (TREM)-1, is predicted to remove the current subjectivity and uncertainty in the diagnosis and therapeutic monitoring of infection and sepsis. PMID- 15833050 TI - Basic principles of real-time quantitative PCR. AB - Real-time quantitative PCR allows the sensitive, specific and reproducible quantitation of nucleic acids. Since its introduction, real-time quantitative PCR has revolutionized the field of molecular diagnostics and the technique is being used in a rapidly expanding number of applications. This exciting technology has enabled the shift of molecular diagnostics toward a high-throughput, automated technology with lower turnaround times. This article reviews the basic principles of real-time PCR and describes the various chemistries available: the double stranded DNA-intercalating agent SYBR Green 1, hydrolysis probes, dual hybridization probes, molecular beacons and scorpion probes. Quantitation methods are discussed in addition to the competing instruments available on the market. Examples of applications of this important and versatile technique are provided throughout the review. PMID- 15833051 TI - Quantitative real-time PCR for cancer detection: the lymphoma case. AB - Advances in the biologic sciences and technology are providing molecular targets for diagnosis and treatment of cancer. Lymphoma is a group of cancers with diverse clinical courses. Gene profiling opens new possibilities to classify the disease into subtypes and guide a differentiated treatment. Real-time PCR is characterized by high sensitivity, excellent precision and large dynamic range, and has become the method of choice for quantitative gene expression measurements. For accurate gene expression profiling by real-time PCR, several parameters must be considered and carefully validated. These include the use of reference genes and compensation for PCR inhibition in data normalization. Quantification by real-time PCR may be performed as either absolute measurements using an external standard, or as relative measurements, comparing the expression of a reporter gene with that of a presumed constantly expressed reference gene. Sometimes it is possible to compare expression of reporter genes only, which improves the accuracy of prediction. The amount of biologic material required for real-time PCR analysis is much lower than that required for analysis by traditional methods due to the very high sensitivity of PCR. Fine-needle aspirates and even single cells contain enough material for accurate real-time PCR analysis. PMID- 15833052 TI - Immunologic markers in the neonatal period: diagnostic value and accuracy in infection. AB - Diagnosis of early-onset neonatal infection has led to the development of several screening tests including C-reactive protein, a very commonly used marker, and cytokines (mainly interleukin-6 and -8), alone or in combination with C-reactive protein, based on the premise that their increases in response to infection may precede that of C-reactive protein. In recent years the search for diagnostic tests has turned to procalcitonin, a propeptide of calcitonin, which appears to be a promising marker of infection in newborn infants. Additionally, specific leukocyte cell surface antigens (mainly CD11b and CD64), detected by flow cytometry, are evaluated as markers of neonatal infection, since their expression on the cell membrane increases in substantial quantities after leukocyte activation by bacteria or their cellular products. This review aims to examine the role of these newly available immunologic indices and to assess their validity as diagnostic markers of infection during the neonatal period. PMID- 15833053 TI - Application of automation and information systems to forensic genetic specimen processing. AB - During the last 10 years, the introduction of PCR-based DNA typing technologies in forensic applications has been highly successful. This technology has become pervasive throughout forensic laboratories and it continues to grow in prevalence. For many criminal cases, it provides the most probative evidence. Criminal genotype data banking and victim identification initiatives that follow mass-fatality incidents have benefited the most from the introduction of automation for sample processing and data analysis. Attributes of offender specimens including large numbers, high quality and identical collection and processing are ideal for the application of laboratory automation. The magnitude of kinship analysis required by mass-fatality incidents necessitates the application of computing solutions to automate the task. More recently, the development activities of many forensic laboratories are focused on leveraging experience from these two applications to casework sample processing. The trend toward increased prevalence of forensic genetic analysis will continue to drive additional innovations in high-throughput laboratory automation and information systems. PMID- 15833054 TI - Applications of molecular methods for Leishmania control. AB - This article reviews the recent advances made in the field of human leishmaniasis. Special emphasis is placed upon the application of various molecular tools for accurate and rapid diagnosis, understanding the mechanisms of drug resistance and identification of vaccine candidates. The focus will be on the major role played by recombinant antigens in the immunoserodiagnosis and progress of the Leishmania genome project, which has enabled researchers to design better PCR primers and molecular probes for microarrays. A special interest is placed on the recombinant antigen (rK39) cloned from the Leishmania chagasi kinesin gene and a very recently cloned recombinant antigen (KE16) from the Old World Leishmania donovani species with high sensitivity and specificity. Advances made in the specific PCR primer designed to diagnose and differentiate various species and strains of Leishmania causing visceral and post-kala-azar dermal leishmaniasis have been covered. Molecular methods (e.g., DNA and protein microarrays) applied to understanding the pathobiology of the parasite, mechanism of host invasion, drug interaction and drug resistance to develop effective therapeutic molecules, gene expression profiling studies that have opened doors to understand many host-parasite relations, effective therapy and vaccine candidates are extensively covered in this review. PMID- 15833055 TI - Assessment of pharmacology and potential anti-obesity properties of H3 receptor antagonists/inverse agonists. AB - Histamine is a key neurotransmitter that alters central nervous system functions in both behavioural and homeostatic contexts through its actions on the histamine (H) subreceptors H(1), H(2) and H(3) G-protein-coupled receptors. H(3)receptors have a diverse central nervous system distribution where they function as both homo- and hetero-receptors to modulate the synthesis and/or release of several neurotransmitters. H(3) receptors are constitutively active, which implies that antagonists of H(3) receptors may also function as inverse agonists to alter the basal state of the receptor and uncouple constitutive receptor-G-protein interactions. Reference H(3) antagonists such as thioperamide and ciproxifan, administered either centrally or systemically, have been shown to cause changes in food consumption and/or body weight in proof-of-concept studies. More recently, several non-imidazole-based H(3) antagonists/inverse agonists have also been described with efficacy in at least one animal model of human obesity. Considerable preclinical effort remains necessary before such compounds achieve therapeutic success or failure. Moreover, ongoing research in a number of laboratories has shed new insights into the effects of H(3) ligands in the control of feeding, appetite and body weight, which offer different results and conclusions. The goal of this review is to appraise these findings and forecast whether any H(3) antagonists/inverse agonists will provide clinical utility to treat human obesity. PMID- 15833056 TI - Advances in anti-obesity therapeutics. AB - Obesity is rapidly becoming an epidemic in developed countries. Currently available anti-obesity therapeutics are only modestly effective and are accompanied by significant adverse effects. In recent years, substantial advances have been made in the basic understanding of brain control of feeding behaviour and metabolism. As a result, several compounds have progressed to Phase III development, with additional compounds at various stages of Phase II development. Most of the late-stage development candidates are CNS agents, which reflects the consensus that the brain exerts a dominant control on feeding behaviour and peripheral metabolism through the autonomic nervous system. Homeostatic mechanisms encompassing hypothalamic/brainstem pathways have long been recognised in obesity research. In addition, non-homeostatic mechanisms encompassing the reward circuit and volitional control need to be targeted to control feeding behaviour and physical activity, especially in humans. While recognising the importance of CNS control, certain peripherally acting agents can affect mitochondrial metabolism, lipolysis, nutrient absorption or the vagal feedback pathway, such that these peripherally acting agents can potentially be combined with CNS agents to achieve maximal efficacy. It is expected that newer generations of anti-obesity therapeutics will be superior to existing agents and will facilitate lifestyle modification. PMID- 15833057 TI - Parathyroid hormone and leptin--new peptides, expanding clinical prospects. AB - There are three injectable and one oral bone-building (i.e., bone anabolic) parathyroid hormone (PTH) peptides. One of the four, Lilly's injectable teriparatide (Forteo), is currently being used, and the other three are in clinical trials. They are being used or assessed only for treating postmenopausal osteoporosis. However, their potential clinical targets now extend far beyond osteoporosis. They can accelerate the mending of even severe non-union fractures; they will probably be used to strengthen the anchorage of pros-theses to bone; they have been shown to treat psoriasis that has resisted other treatments; they can increase the size of haematopoietic stem cell proliferation and accelerate the endogenous repopulation or repopulation by donor transplants of bone marrow depleted by chemotherapeutic drugs; and they may prevent vascular ossification. Leptin, a member of the cytokine superfamily has a PTH-like osteogenic activity and may even partly mediate PTH action. But leptin has two drawbacks that cloud its therapeutic future. First, apart from directly stimulating osteoblastic cells, it targets cells in the hypothalamic ventromedial nuclei and through them it reduces oestrogenic activity by promoting osteoblast-suppressing adrenergic activity. Second, it stimulates vascular and heart valve ossification, which leads to such events as heart failure and diabetic limb amputations. PMID- 15833058 TI - Emerging and potential therapies for osteoporosis. AB - Osteoporotic fractures are an important public health problem, contributing substantially to morbidity and mortality in an ageing world population and consuming considerable health resources. Currently available pharmacological therapies for prevention of fragility fractures are limited in scope, efficacy and acceptability to patients. Considerable efforts are being made to develop new, more effective treatments for osteoporosis and to refine/optimise existing therapies. These novel treatments include an expanding array of drugs that primarily inhibit osteoclastic bone resorption; oestrogenic compounds, bisphosphonates, inhibitors of receptor activator of nuclear factor-kappaB ligand signalling, cathepsin K inhibitors, c-src kinase inhibitors, integrin inhibitors and chloride channel inhibitors. The advent of intermittent para-thyroid hormone (PTH) therapy has provided proof-of-principle that osteo-blast-targeted (anabolic) agents can effectively prevent osteoporotic fractures, and is likely to be followed by the introduction of other therapies based upon PTH, such as orally active PTH analogues, antagonists of the calcium sensing receptor, PTH related peptide analogues, and/or agents that induce osteoblast anabolism via pathways involving key, recently identified, molecular targets (wnt low-density lipoprotein receptor-related protein-5 signalling, sclerostin and matrix extracellular phosphoglycoprotein). PMID- 15833059 TI - Agents in development for the treatment of diabetic nephropathy. AB - Diabetic nephropathy is a leading cause of end-stage renal disease, and accounts for significant morbidity and mortality in patients with diabetes. Diabetic nephropathy seems to occur as a result of an interaction between metabolic and haemodynamic factors, which activate common pathways that lead to renal damage. In the past, the treatment of diabetic nephropathy has focused on the control of hyperglycaemia. Newer targets, some of which are linked to glucose-dependent pathways, appear to be a major focus of new treatments directed against the development and progression of renal damage as a result of diabetes. It is anticipated that additional therapeutic approaches that inhibit both metabolic and haemodynamic pathways will include strategies that target growth factors, cytokines and intracellular second messengers. Such an approach is expected to lead to improved therapies for the treatment of diabetic nephropathy. PMID- 15833060 TI - Novel therapies in transplantation. AB - Immunosuppressant drugs are crucial in order to protect transplanted kidney, heart or liver against the body's natural defence mechanisms. These drugs are used to prevent both acute rejection of the organ as well as chronic deterioration of the graft over longer periods of time. Currently used immunosuppressant drugs are calcineurin inhibitors (cyclosporin and tacrolimus), steroids (prednisolone and methylprednisolone), antimetabolites (azathioprine and mycophenolate mofetil), antiproliferatives (sirolimus) and monoclonal antibodies that are used as an induction therapy (basilximab, daclizumab and muromonab). Over time, we have learnt more about existing immunosuppressive choices and the ways to monitor these drugs, and the current trend in immunosuppression therapy is towards tailoring the therapy according to an individual patient. The major issue that is now emerging is not how to prevent acute rejection, as current drugs are all efficacious, but rather how to avoid the long-term side effects that can harm both the graft and host, and negatively influence compliance. The market in transplantation is considerable and still growing; the whole market including the immuno-suppressants used in autoimmune diseases had sales of approximately pound 1.5 billion in 2001. There are currently 23 new drugs in advanced clinical development intended to be used in either organ transplantation or as treatment of autoimmune diseases. The new drugs that are intended to be used in transplantation are mostly analogues of currently used drugs with improved safety and pharmaceutical profiles. PMID- 15833061 TI - Antiviral therapy of influenza. AB - Influenza viruses are a leading cause of morbidity and mortality worldwide despite the availability of an effective vaccine. The emergence of highly pathogenic avian influenza viruses in southeast Asia, which can infect and kill humans and for which there is no vaccine, has heightened the need to establish a supply of effective antivirals. Two effective classes of anti-influenza drugs are currently available, and many attractive targets exist for further development. This review presents the current status of antiviral therapy of, and highlights the challenges presented by, the threat of pandemic influenza. PMID- 15833062 TI - Inhibitors of the mammalian target of rapamycin. AB - The mammalian target of rapamycin (mTOR) is a downstream protein kinase of the phosphatidylinositol 3'-kinase-Akt signalling pathway. As a result of its position within this pathway and its central role in controlling cellular growth, mTOR is viewed as an important target for anticancer therapeutics development. Currently, the mTOR inhibitor rapamycin (sirolimus, Wyeth) and its derivatives temsirolimus (CCI-779, Wyeth), everolimus (RAD-001, Novartis Pharma AG) and AP 23573 (Ariad Pharmaceuticals) are being evaluated in cancer clinical trials. Preclinical studies suggest that sensitivity to mTOR inhibition may correlate with aberrant activation of the phosphatidylinositol 3'-kinase pathway and/or with aberrant expression of cell-cycle regulatory or antiapoptotic proteins. Clinical trial results show that mTOR inhibitors are generally well tolerated and may induce prolonged stable disease and even tumour regressions in a subset of patients. Questions remain regarding optimal dose, schedule, patient selection and combination strategies for this novel class of agents. PMID- 15833063 TI - Lucinactant: a novel synthetic surfactant for the treatment of respiratory distress syndrome. AB - Lucinactant (Surfaxin, Discovery Laboratories) is a synthetic surfactant, which contains the novel peptide, sinapultide, a surfactant-associated protein B mimic. Randomised clinical trials suggest that this compound is a safe and effective treatment for respiratory distress syndrome in preterm infants. It is also being actively investigated for other indications, including meconium aspiration syndrome, treatment of bronchopulmonary dysplasia in neonates, acute respiratory distress syndrome and asthma. A novel aerosol formulation administered with nasal continuous positive airway pressure is also under development for treatment of respiratory insufficiency in neonates. Its non-animal origin may make it an attractive alternative to present animal-derived surfactants by eliminating the risks of infection and immunogenicity related to the latter. PMID- 15833064 TI - The benefits of abciximab questioned in percutaneous coronary interventions following pretreatment with high-dose clopidogrel. PMID- 15833065 TI - Will the new CB1 cannabinoid receptor antagonist SR-147778 have advantages over rimonabant? PMID- 15833066 TI - Advances in anti-viral therapeutics. AB - Hepatitis C virus (HCV) and HIV infections are of high economical importance as they have a global impact, high mortality rates and, consequently novel treatments are urgently needed. Worldwide, 170 and 34 - 46 million people are infected with HCV and HIV, respectively. HCV and HIV medications constitute the largest antiviral markets today. The efforts in fighting HCV infections are still limited to early clinical development, trying to establish proof-of-concept in man. However, in HIV, there is a better chance that some of the new drugs on new and old targets (indirect and direct antiviral targets, like CCR5 and HIV protease, respectively) will finally reach the market. PMID- 15833067 TI - Cell therapies for neurological disease--from bench to clinic to bench. AB - The lack of any meaningful regeneration in the adult central nervous system (CNS) subsequent to damage or degeneration stimulated the concept of replacement of the deficient cells by transplantation. Thus, much time and effort has been spent on investigating the potential of cell replacement therapy for repair in a range of conditions of the CNS over the last 25 years. As promising proof of principle basic science results were slowly converted to success in clinical transplantation trials in Parkinson's disease (PD), the future seemed very encouraging for cell therapy. However, the recent randomised, double-blind, placebo-controlled studies of fetal neural transplantation in PD have produced more equivocal results, which has dampened enthusiasm for this approach. However, whilst the translation of cell therapies to the clinic is in limbo, the emergence of stem cells as a source of the replacement tissue has revitalised the laboratory-based studies. This paper attempts to reconcile these disparate views and put forward the authors' view on the future of this form of biological therapy and its implications for related therapies. PMID- 15833068 TI - Immunomodulatory effects of human recombinant granulocyte-macrophage colony stimulating factor (rhuGM-CSF): evidence of antitumour activity. AB - Human recombinant granulocyte-macrophage colony-stimulating factor (rhuGM-CSF) is traditionally used as supportive care for patients undergoing cytotoxic chemotherapy or haematopoietic cell progenitor mobilisation. Emerging evidence suggests rhuGM-CSF, through activity on monocytes and dendritic cells, acts as a potent modulator of immune responses and has the ability to recruit inflammatory cells and cytokines to local and systemic sites of infection. The immunomodulatory effects of rhuGM-CSF suggest the potential to enhance innate and acquired immune responses against tumour-related antigens. Enhancement of innate antitumour immunity, especially in the context of minimal residual disease, is of central importance and presents the potential for meaningful contributions to long-term disease survival. This article discusses the immunomodulatory effects of rhuGM-CSF in the context of single-agent therapy in solid tumours, as well as combination therapy in lymphoma. In addition, dendritic cell modulation with rhuGM-CSF in haematopoietic progenitor grafts and rhuGM-CSF-transduced tumour vaccines will be discussed. PMID- 15833069 TI - Involvement of ICAM-1 in bone metabolism: a potential target in the treatment of bone diseases? AB - Bone diseases such as osteoporosis, osteoarthritis and rheumatoid arthritis (RA) affect a great proportion of individuals, with debilitating consequences in terms of pain and progressive limitation of function. Existing treatment of these pathologies has been unable to alter the natural evolution of the disease and, as such, a clearer understanding of the pathophysiology is necessary in order to generate new treatment alternatives. One therapeutic strategy could involve the targeting of intercellular adhesion molecule-1 (ICAM-1; CD54). In bone, ICAM-1 is expressed at the surface of osteoblasts (Obs) and its counter-receptor, leukocyte function-associated antigen-1 (LFA-1; CD11a), at the surface of osteoclast (Oc) precursors. ICAM-1 blockade between the Ob and the pre-Oc results in an inhibition of Oc recruitment and a modulation of inflammation, which could potentially help in controlling disease activity in bone pathologies. So far, clinical studies on ICAM-1 blockade in bone diseases have been limited to RA. A better understanding of the implication of this adhesion molecule in Ob/Oc interactions and inflammatory mediation in the bone pathological state, however, is needed. As new discoveries on the role of this adhesion molecule are being reported, ICAM-1 could become a potential target for other bone diseases as well. PMID- 15833070 TI - Therapeutic role of CD8+ T cells in HIV-1 infection: targets and suppressors of viral replication. AB - CD8+ T cells are pivotal in controlling viral replication in HIV-1-infected subjects. However, in chronic infection, HIV-1-specific CD8+ T cells fail to adequately control infection, presenting incomplete maturation and more severe functional impairment with advanced disease. Accumulating evidence has shown that CD8+ T cells can also be productively infected by HIV-1. Whether HIV-1 infection of CD8+ T lymphocytes impacts on their antiviral activity remains to be determined. This review explores the potential mechanisms of HIV-1 infection of CD8+ T cells, its likely contribution to the immunopathogenesis of HIV-1 infection and potential therapeutic interventions. PMID- 15833071 TI - Gene therapy for chronic obstructive pulmonary disease: twilight or triumph? AB - Chronic obstructive pulmonary disease (COPD) is a clinical syndrome presenting as progressive airflow limitation that is poorly reversible as a result of bronchitis and emphysema. The prevalence of COPD is alarming and even more so its current and projected impact on morbidity and mortality. To date, there are no effective treatments for emphysema, nor are there efficient clinical management strategies. Existing and prospective therapies, although promising, have yet to demonstrate their efficacy to slow, halt or reverse the disease. Novel approaches using gene therapy and stem cell technologies may offer new opportunities. However, this will remain almost entirely dependent on a more thorough understanding of the pathogenesis of COPD. This review is not aimed at highlighting the vast effort of studying COPD, but rather describing the state of the field in an abstract fashion to expose the focus of research efforts to date, which has primarily been limited to predisposing factors and inflammation. We would like to draw attention to other elements of the disease, such as the alveolar remodelling that characterises emphysema. Although the main cause may prove to be elusive, carefully designed clinical treatment and management may deliver the required therapeutic outcome. PMID- 15833072 TI - Gene therapy for obesity. AB - A sedentary life-style and an environment of caloric abundance have contributed to the recent rise in the incidence of obesity. Treating such a complex disease requires an understanding of its underlying molecular mechanisms that has only recently become possible with the sequencing of the human genome and the mapping of hundreds of genes associated with increased risk of obesity. With few safe and efficacious weight-maintenance drugs available on the market, gene therapy offers an alternative long-term treatment modality. Still in its infancy, gene therapy for obesity is poised for significant progress, due in large part to a wide variety of available gene targets and the development of novel systems to control gene expression. Coincidently, novel vectors are being developed based on adeno associated virus providing efficient and sustained expression of a transgene in metabolically important tissues. These advances are driving the development of gene therapy as a viable therapeutic option in treating obesity and its associated disorders. PMID- 15833073 TI - siRNA-mediated inhibition of angiogenesis. AB - Small interfering RNA (siRNA) has rapidly become the agent of choice for gene function analysis through loss-of-function phenotypes. Especially in complicated (patho)physiological processes such as angiogenesis, where vast numbers of proteinaceous factors are involved, the siRNA application allows relatively fast analysis of pathways and identification of new target genes. The first studies on the therapeutic effects of siRNA in angiogenesis show that this new 'drug' class holds great promise for therapeutic intervention. Two strategies emerge: the use of unmodified or the use of complexed, targeted and/or protected nucleic acids. The challenge for clinical application will be to control off-target effects and the transient character of the sequence-specific silencing effect, and to address the targeted delivery to the cell types involved in the various stages of angiogenesis. This is especially important as clinical studies indicate a profound heterogeneity of the angiogenic vasculature. PMID- 15833074 TI - The role of reverse genetics in the development of vaccines against respiratory viruses. AB - Despite their significance, the only available vaccines against respiratory viruses are those for the prevention of influenza. Attempts have been made to produce vaccines against other respiratory viruses using traditional techniques, but have met with little success. Reverse genetics, although still a relatively new tool for the manipulation of negative-strand RNA viruses, has great potential for the preparation of vaccines against many of the common respiratory viruses. In the preparation of live vaccines, reverse genetics systems allow the direct modification of the specific regions in the genomes of negative-stranded RNA viruses concerned with attenuation; the ultimate goal is the introduction of site specific mutations through a cDNA intermediate in order to develop strains with the requisite attenuation, antigenic and growth properties needed in a vaccine. These techniques can also be used to disarm potentially highly pathogenic viruses, such as emerging H5N1 avian influenza viruses, in order to facilitate large-scale preparation of viruses for use in inactivated vaccines under conditions of manufacturing safety. Before these vaccines become available, residual issues concerned with intellectual property rights to the technology and its application will need to be resolved. PMID- 15833075 TI - Contraceptive vaccines targeting sperm. AB - Overpopulation is a global problem of significant magnitude, with grave implications for the future. Development of new contraceptives is necessary, as existing forms of birth control are unavailable, impractical and/or too expensive for many individuals due to sociological, financial or educational limitations. Immunocontraception and, in particular, the targeting of antibodies to sperm specific antigens implicated in sperm-egg binding and fertilisation offers an attractive approach to control fertility. Sperm-specific antibodies may impair fertility by inhibiting sperm motility, by reducing penetration of the cervical mucus by sperm, or by interfering in sperm capacitation or the acrosome reaction; alternatively, antisperm antibodies may invoke the complement cascade, resulting in sperm lysis. The antibodies raised against sperm-specific antigens have proved to be extremely effective at reducing sperm-egg interactions in vitro; fertility trials in subhuman primates will eventually be needed to prove the effectiveness of the sperm antigens in terms of contraceptive efficacy before trials in humans can be justified. In addition, existing and emerging strategies (such as sperm proteomics, the determination of molecular and structural details of sperm proteins, and the modelling of protein-ligand interactions using X-ray and/or NMR structures to name a few) are expected to provide the experimental foundation for the design of small molecule inhibitors with antifertility effects. The technology underpinning vaccine development is constantly being developed and the introduction of DNA/RNA vaccines is certain to impact upon the field of immunocontraception. PMID- 15833076 TI - Efalizumab: continuous therapy for chronic psoriasis. AB - Efalizumab is a humanised monoclonal antibody targeting the CD11a subunit of lymphocyte function-associated antigen-1, specifically developed for psoriasis. Indicated for patients with moderate-to-severe plaque psoriasis, efalizumab is FDA-approved in the US for patients aged > or = 18, as well as being approved in several other European countries. Clinical studies have proven the efficacy of efalizumab for a majority of patients, improving quality of life with continuous maintenance therapy by means of weekly subcutaneous self-injections. Controlled trials have demonstrated the safety and tolerability of efalizumab. Clinical trials have established that approximately 30% of patients can achieve a PASI-75 response within 12 weeks of initiating treatment, with further clinical benefit noted with continued therapy up to 24 and even 36 months of therapy. Efalizumab may thus potentially offer patients a safe option for long-term safe control in managing their disease. PMID- 15833077 TI - Infliximab for the treatment of early rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a systemic, chronic inflammatory disease characterised predominately by polyarthritis with frequent progression to permanent joint damage and disability. Evidence shows that starting treatment with disease-modifying antirheumatic drug (DMARD) therapy early in the course of disease can slow radiographic progression of RA compared with a more delayed approach. Moreover, in the early stages of RA, treatment with a combination of methotrexate (MTX), a traditional DMARD, and a biologic with tumour necrosis factor (TNF)-alpha blocking activity has proven to be more effective than using MTX alone. Among the approved TNF-alpha inhibitors, infliximab is a chimeric monoclonal antibody with high affinity and specificity for its target cytokine. It binds to soluble TNF monomers and trimers, as well as membrane-bound TNF alpha, forming a stable complex which prevents TNF-alpha from binding to its receptor and triggering a biological response. The combination of MTX and infliximab therapy has shown superior clinical outcomes compared with MTX monotherapy in early RA, as well as greater protection against joint damage and physical disability. Although infliximab therapy has been associated with side effects, including serious infections, this drug can be administered with an acceptable margin of safety for several years by appropriate selection of patients, screening for latent and active tuberculosis, and monitoring of patients for infection and other toxicities. The knowledge of the benefits of infliximab therapy for early RA affords groundwork for developing more effective treatment strategies that can minimise disease progression over the long term. PMID- 15833078 TI - Mathematical models of oxygen and carbon dioxide storage and transport: the acid base chemistry of blood. AB - This article describes a mathematical model of the acid-base chemistry of blood. The model is formulated from first principles by considering the "components" of blood and the reaction equations in the plasma and erythrocyte fractions. Equations are formulated to describe the total concentration of blood components, the physicochemical properties, and the equilibrium position of reactions. The model includes 28 equations and 12 parameters. All equations can be solved from six variables included in the model. The model uses simple mathematics, without introducing intermediate concepts or linear coefficients necessary for algebraic solution. Model equations are solved simultaneously using numerical methods. Model parameters are estimated and the model verified for plasma, fully oxygenated blood, and deoxygenated blood. Published data are used to estimate model parameters and normal conditions and to verify model simulations. The model reproduces experimental results, including addition or removal of CO2, or strong acid to plasma; CO2, strong acid or haemoglobin to blood; and the effects of deoxygenating blood. The model can also be used as the basis for models of whole body CO2 transport as illustrated in the accompanying article. As such, it is possible to simulate the effects on blood of physiological changes in ventilation or metabolism. PMID- 15833079 TI - Mathematical models of oxygen and carbon dioxide storage and transport: interstitial fluid and tissue stores and whole-body transport. AB - This article describes a mathematical model of whole-body O2 and CO2 transport. The model includes representation of the acid-base chemistry of the blood, interstitial fluid, and tissues, plus transport of O2 and CO2 between compartments representing tissues, interstitial fluid, arterial and venous blood, and lungs. The model includes equations for calculation of all concentrations in the compartments, including equations describing the physicochemical properties and reaction equations of interstitial fluid and tissues. In addition, the model includes equations that describe the flow of substrate between the compartments and differential equations allowing calculation of the changes in state variables caused by the flow of substrates between the compartments. This model is designed to calculate the effects of metabolic and respiratory perturbations, such as variation in breathing pattern or production of strong acid at the tissues. The model reproduces the results of published experiments when used to simulate (1) normal conditions in the lungs, arterial and venous blood, interstitial fluid, and tissues during normal ventilation; (2) the characteristic two-exponential response to changes in minute ventilation; and (3) the relationship between arterial blood values of PCO2 and HCO3,p during inspiration of different fractions of CO2. PMID- 15833080 TI - Dendritic epidermal T-cell activation. AB - Although gammadelta T cells compose a small proportion of lymphocytes in lymphoid compartments and peripheral blood, they are the major T-cell population present in epithelial tissues. However, the role played by gammadelta TCR expressing intraepithelial lymphocytes (IEL) has been enigmatic. The location of tissue resident IEL suggests that they are important members of the first line of defense against insult for organs exposed to the environment, including the skin, gut, lungs, and reproductive system. Dendritic epidermal T cells (DETC) are the skin-resident gammadeltaIEL and serve as a model system for gammadeltaIEL in other locations. DETC have demonstrated importance in the modulation of immune responses, surveillance and repair of tissue, and resistance to infection. This work discusses recent developments in understanding DETC activation. PMID- 15833081 TI - Properties of glycolipid-enriched membrane rafts in antigen presentation. AB - Presentation of antigen to T cells represents one of the central events in the engagement of the immune system toward the defense of the host against pathogens. Accordingly, understanding the mechanisms by which antigen presentation occurs is critical toward our understanding the properties of host defense against foreign antigen, as well as insight into other features of the immune system, such as autoimmune disease. The entire antigen-presentation event is complex, and many features of it remain poorly understood. However, recent studies have provided evidence showing that glycolipid-enriched membrane rafts are important for efficient antigen presentation; the studies suggest that one such function of rafts is trafficking of antigen-MHC II complexes to the presentation site on the surface of the antigen-presenting cell. Here, we present a critical discussion of rafts and their proposed functions in antigen presentation. Emerging topics of rafts and antigen presentation that warrant further investigation are also highlighted. PMID- 15833082 TI - The caspase-8 modulator c-FLIP. AB - Death receptors belong to the tumor necrosis factor receptor superfamily and can induce apoptosis through activation of procaspase-8. The cellular FLICE inhibitory protein (c-FLIP) is able to modulate activation of procaspase-8 and thereby prevents induction of apoptosis mediated by death receptors. As an important modulator of caspase-8, c-FLIP regulates life and death in various types of normal cells and tissues, such as lymphoid cells, and renders resistance to death receptor-mediated apoptosis in many types of cancer cells. In addition to an apoptosis modulator, c-FLIP has been shown to exert other physiological functions related to cell proliferation and tumorigenesis. Dysregulation of c FLIP expression has been shown to be associated with various diseases, such as cancer and autoimmune diseases, and c-FLIP might be a critical target for therapeutic intervention. This review focuses on recent findings about the physiological function and intracellular regulation of c-FLIP. PMID- 15833083 TI - Antigen-driven T-cell repertoire selection. AB - The immune system has evolved to combat infectious organisms through the coordination of both innate and acquired immune responses. Innate immune responses generally pass their roles in eradicating pathogens to acquired immune responses conducted by T and B cells. T and B cells generate diverse antigen specific receptors through gene recombination, and both cell repertoires are shaped during their development by recognizing self-antigens. After cell maturation, the repertoire selection is further developed by recognizing non-self antigens, which is essential for efficient eradication of pathogens. Those antigen-driven selections are seen not only in conventional CD4(+) and CD8(+) T cells but also in CD4(+)CD25(+) regulatory T cells. In this article, we review current concepts of repertoire selection of conventional T cells, as well as regulatory T cells, and discuss the physiological role of such selection in controlling acquired immune responses against pathogens. PMID- 15833086 TI - Migraine pathophysiology. AB - A combination of basic science and human physiology, particularly functional neuroimaging, has radically altered our understanding of migraine with a focus on brain mechanisms for this common and disabling disorder. Genetic studies have begun to provide plausible targets for the basic molecular defect in terms of ion channels, albeit thus far in the rare condition of familial hemiplegic migraine (FHM). Migraine pathophysiology involves the trigeminovascular system and central nervous system modulation of the pain-producing structures of the cranium. The degree to which head pain results from the activation of the nociceptors of pain producing intracranial structures, or to the facilitation or lack of inhibition of afferent signals, is not clear at this time. An understanding of the pain mechanism is likely to provide insights into the mechanisms underlying the more generalized sensory dysfunction that is so typical of migraine. PMID- 15833084 TI - Activated STAT5 proteins induce activation of the PI 3-kinase/Akt and Ras/MAPK pathways via the Gab2 scaffolding adapter. AB - The active forms of STAT5A (signal transducer and activator of transcription 5A) and STAT5B are able to relieve the cytokine dependence of haematopoietic cells and to induce leukaemia in mice. We have demonstrated previously that activation of the PI3K (phosphoinositide 3-kinase) signalling cascade plays a major role in cell growth and survival induced by these proteins. Interaction between STAT5 and p85, the regulatory subunit of the PI3K, has been suggested to be required for this activation. We show in the present study that the scaffolding protein Gab2 [Grb2 (growth-factor-receptor-bound protein 2)-associated binder-2] is an essential component of this interaction. Gab2 is persistently tyrosine phosphorylated in Ba/F3 cells expressing caSTAT5 (constitutively activated STAT5), independent of JAK2 (Janus kinase 2) activation where it interacts with STAT5, p85 and Grb2, but not with Shp2 [SH2 (Src homology 2)-domain-containing tyrosine phosphatase] proteins. Interaction of STAT5 with Gab2 was also observed in Ba/F3 cells stimulated with interleukin-3 or expressing the oncogenic fusion protein Tel-JAK2. The MAPKs (mitogen-activated protein kinases) ERK1 (extracellular-signal-regulated kinase 1) and ERK2 were constitutively activated in the caSTAT5-expressing cells and were found to be required for caSTAT5-induced cell proliferation. Overexpression of Gab2-3YF, a mutant of Gab2 incapable of binding PI3K, inhibited the proliferation and survival of caSTAT5-expressing cells as well as ERK1/2 and Akt/protein kinase B phosphorylation. Taken together, our results indicate that Gab2 is required for caSTAT5-induced cell proliferation by regulating both the PI3K/Akt and the Ras/MAPK pathways. PMID- 15833087 TI - Brain hyperexcitability: the basis for antiepileptic drugs in migraine prevention. AB - Abnormal brain excitability may provide the susceptibility for triggering migraine attacks. Antiepileptic drugs may diminish neuronal excitability and consequently reduce the frequency of migraine. Because migraine aura is predominantly visual, hyperexcitability of the occipital cortex has been the focus of investigations. Functional magnetic resonance imaging of the brain and magnetoencephalography provide the most consistent evidence for the role of brain hyperexcitability in migraine and confirm that triggering an abnormal electric and metabolic event consistent with the cortical spreading depression (CSD) of Leao is anatomically and functionally linked with migraine aura symptoms. Future drug discovery should focus on the interface between the excitable cell and the earliest events of CSD. PMID- 15833088 TI - Migraine: epidemiology, impact, and risk factors for progression. AB - Migraine is a chronic and sometimes progressive disorder characterized by recurrent episodes of headache and associated symptoms. This article reviews the epidemiology of and the risk factors for migraine described in population studies, and discusses the burden of disease and the socioeconomic costs of migraine. In the years prior to puberty, migraine is more common among boys than girls. By the onset of puberty, migraine is more prevalent in girls, and by the late teens, females are about twice as likely to suffer from migraine as males. The prevalence of migraine peaks in both sexes during the most productive years of adulthood (age 25 to 55 years) and, in the United States, the prevalence is higher in individuals of lower socioeconomic status. Direct costs of migraine include the cost of migraine medications and health care expenses. Indirect costs associated with migraine include reduced productivity due to absenteeism and reduced performance while at work. Recent evidence suggests that a subgroup of migraine patients may have a clinically progressive disorder. Future epidemiologic studies should focus on identifying patients who are at higher risk for progression and on assessing the impact of intervention strategies on disease progression. PMID- 15833089 TI - General principles of migraine management: the changing role of prevention. AB - The role of preventive treatment in the management of migraine must be reassessed. Many patients who would benefit from preventive therapy do not receive it, and many might benefit from earlier and more aggressive use of preventive therapy. Physicians who treat migraine have long recognized that a subset of patients with episodic migraine evolve to chronic forms of the disorder that are difficult to treat and have a poor outcome. This article reviews the evidence for current and emerging prophylactic migraine treatment and raises the possibility that timely use of prophylactic treatment might modify or prevent the transformation to chronic migraine and the extreme disability that characterizes a small but significant subset of the migraine population. Along with aggressive treatment of the acute pain and other symptoms of migraine, prevention of progression to severe forms of this disorder will increasingly be a focus and goal of the treatment. PMID- 15833090 TI - Molecular pharmacology of topiramate: managing seizures and preventing migraine. AB - Topiramate is a neuromodulatory compound with stabilizing properties that was initially introduced for the management of partial seizures. Topiramate has been demonstrated to modify several receptor-gated and voltage-sensitive ion channels, including voltage-activated Na+ and Ca2+ channels and non-NMDA receptors. These receptors have been implicated in the pathophysiology of both epilepsy and migraine. The pharmacological mechanisms of action for topiramate that may explain its antiepileptic and migraine preventive activities will be discussed in this review. In addition, the potential relationship between the molecular activities of topiramate and its efficacy in epilepsy and migraine prevention will be emphasized. PMID- 15833091 TI - Topiramate in migraine prevention. AB - The efficacy of topiramate in migraine prevention (prophylaxis) was established in two multicenter, randomized, double-blind, placebo-controlled, pivotal trials. Topiramate has received regulatory approval for use in adults for migraine prophylaxis (prevention) in the US and numerous other countries, including France, Ireland, Switzerland, Brazil, Taiwan, Spain, and Australia. Treatment with 100 or 200 mg per day of topiramate was associated with significant reductions in the frequency of migraine headaches, number of migraine days, and use of acute medications. No increase in efficacy was observed between 100 and 200 mg per day of topiramate. Based on efficacy and tolerability, 100 mg per day of topiramate should be the initial target dose for most patients. The most common adverse events were paresthesia, fatigue, decreased appetite, nausea, diarrhea, weight decrease, and taste perversion. Topiramate is a first-line migraine preventive drug and should especially be considered as a preferred treatment for all patients who are concerned about gaining weight, who are currently overweight, or who have coexisting epilepsy. PMID- 15833092 TI - Practical use of topiramate for migraine prevention. AB - When treating patients with migraine, clinicians should consider prescribing appropriate combinations of acute and preventive therapies. An effective migraine preventive therapy should be prescribed to patients with frequent (> or = 2 migraines per month) or severe migraine. Topiramate has been shown to be an effective and generally well-tolerated migraine prophylaxis (preventive) therapy in adults, as demonstrated in several large, controlled trials. The most common adverse events in these trials were paresthesia, fatigue, anorexia, nausea, taste alteration, and diarrhea. Most adverse events were mild to moderate and transient in nature. Although patients should take migraine-preventive medications for approximately 2 to 3 months before evaluating effect, topiramate has shown efficacy as early as the first month of treatment. This article describes "real world" approaches to using topiramate as a migraine-preventive therapy. Topiramate has received regulatory approval for the prophylaxis of migraine headache in adults in the United States and many other countries. The practical issues discussed in this article will enable clinicians to maximize the effectiveness while minimizing the side effects associated with this preventive agent. PMID- 15833093 TI - Management of chronic daily headache: challenges in clinical practice. AB - Chronic daily headache (CHD) refers to a category of headache disorders that are characterized by headaches occurring on more than 15 days per month. This category is subdivided into long- and short-duration (>4 or <4 hours) CDH disorders based on the duration of individual headache attacks. Examples of long duration CDH include transformed migraine (TM), chronic migraine (CM), new daily persistent headache (NDPH), acute medication overuse headache, and hemicrania continua (HC). The goal of this review is to enable clinicians to accurately diagnose and effectively manage patients with long-duration CDH. Patients with CDH often require an aggressive and comprehensive treatment approach that includes a combination of acute and preventive medications, as well as nondrug therapies. PMID- 15833094 TI - Mood stabilizers: a labile label. PMID- 15833095 TI - Lithium - a continuing story in the treatment of bipolar disorder. AB - OBJECTIVE: To review the literature on the use of lithium in the treatment of bipolar disorder and highlight the evidence base supporting its efficacy and safety. METHOD: A selective literature review. RESULTS: Lithium is widely believed to be effective against acute mania, acute bipolar depression and in relapse prevention to either mania or depression. In fact, the data supporting efficacy in acute treatment are less impressive than is often claimed, whereas for relapse prevention and suicide prevention no other agent has comparable depth of support. Lithium is best described as the bench mark treatment for bipolar disorder, rather than the gold standard, because only a minority of patients show major clinical benefit. There is a developing need for further trials against new alternatives and in combination studies. CONCLUSION: Lithium has a continuing important role in the clinical management of bipolar disorder. Its under utilization in North America reflects opinion rather than evidence and the demonstrated anti-suicide effects should help to reignite interest in its use. PMID- 15833096 TI - Valproate in bipolar disorder: 2000 onwards. AB - OBJECTIVE: To describe the evidence for the efficacy and tolerability of valproate in treatment of bipolar disorders, as well as factors associated with favorable or unfavorable outcomes. METHOD: Studies published from 2000 onwards were reviewed, as well as published abstracts. For clinical trials, randomized, prospective studies were emphasized. RESULTS: Several mechanisms of action of valproate on central nervous system neurons that may be relevant to its actions in bipolar disorders have been recently reported. These include inhibition of glycogen synthase kinase and activation of extracellular signal-regulated kinase. Several of the actions overlap those observed from lithium. Valproate is effective in treatment of mania, and somewhat more effective in certain patient subgroups than other treatments, e.g. mixed mania, and mania with prominent irritability. Valproate is comparable with olanzapine in maintenance treatment, and somewhat better tolerated. Higher serum levels, particularly above 110 microg/ml, are associated with more reports of weight gain, sedation, and reductions in platelet count. Valproate may be associated with an increased rate of polycystic ovarian syndrome, with increased weight contributing to the risk. Valproate reduces total cholesterol levels, particularly among patients with baseline elevations of cholesterol. Several studies indicate that valproate can be beneficially combined with antipsychotic drugs and other treatments for bipolar disorder. CONCLUSION: Valproate continues to be studied in further clarification of its mechanisms, efficacy, risks, and spectrum of benefits in bipolar disorder. It is a major treatment for bipolar disorder, both in monotherapy and combination therapy regimens. PMID- 15833097 TI - Lamotrigine and antiepileptic drugs as mood stabilizers in bipolar disorder. AB - OBJECTIVE: To review the clinical trials literature on the use of antiepileptic drugs (AED) as mood stabilizers and to suggest an evidence-based approach when utilizing these agents in bipolar disorder. METHOD: The literature is reviewed and subdivided into the following sections: carbamazepine and oxcarbazepine, valproate, lamotrigine, gabapentin and other AED, and discussion. RESULTS: Data exist to support the use of carbamazepine and valproate - and to a lesser extent, oxcarbazepine - in the management of acute manic episodes associated with bipolar I disorder. Lamotrigine, gabapentin, and other AED have not demonstrated consistent anti-manic effects. Clinical trials data favor lamotrigine over all other AED in the treatment of acute bipolar I depression and in rapid cycling bipolar disorder (particularly type II), although the absence of an active comparator in these lamotrigine trials must be noted. Lamotrigine, carbamazepine, and valproate all have evidence supporting their roles as potential long-term mood stabilizers to prevent bipolar relapse, with lamotrigine having a stronger effect in the prevention of depression. CONCLUSION: The AED are a heterogeneous group of medications with differential spectrum of efficacy in the treatment of bipolar disorder. PMID- 15833098 TI - Immunologic uniqueness of the genital tract: challenge for vaccine development. AB - Although the genital tract is considered to be a component of the mucosal immune system, it displays several distinct features not shared by other typical mucosal tissues and external secretions. Both male and female genital tract tissues lack inductive mucosal sites analogous to intestinal Peyer's patches. Consequently, local humoral and cellular immune responses stimulated by infections [with e.g. Neisseria gonorrhoeae, Chlamydia trachomatis, papilloma virus, and human immunodeficiency virus (HIV-1)] are weak or absent, and repeated local intravaginal immunizations result in minimal humoral responses. In contrast to typical external secretions such as intestinal fluid that contain secretory immunoglobulin A (S-IgA) as the dominant isotype, semen and cervico-vaginal fluid contain more IgG than IgA. Furthermore, irrespective of the route of infection, humoral immune responses to HIV-1 are dominated by specific IgG and low or absent IgA antibodies in all external secretions. Because a significant proportion of IgG in genital tract secretions is derived from the circulation, systemic immunization may provide protective IgG antibody-mediated immunity in the genital tract. Furthermore, combined systemic and mucosal (oral, rectal, and especially intranasal) immunization may induce protective humoral responses in both the systemic and mucosal compartments of the immune system. PMID- 15833099 TI - Expression of killer immunoglobulin-like receptors on peripheral blood NK cell subsets of women with recurrent spontaneous abortions or implantation failures. AB - PROBLEM: Decidual natural killer (NK) cells express inhibitory receptors (killer immunoglobulin-like receptors, KIRs), which bind to ligands on trophoblast cells (human leucocyte antigen, HLA-C). This interaction appears to block NK cytotoxicity against trophoblast cells. In this study, we investigated the expression of inhibitory and activating receptors in peripheral blood NK cells of women with recurrent spontaneous abortion (RSA) or implantation failures. METHOD OF STUDY: CD56(dim)/CD16(+), CD56(bright)/CD16(-) NK cells and CD56(+)/CD3(+) NKT cells of women with RSA or in vitro fertilization (IVF) failures and normal controls were analyzed for the expression of CD158a, CD158b inhibitory KIRs or CD161-activating receptors, by flow cytometric analysis. RESULTS: CD158a and CD158b inhibitory receptor expression by CD56(dim)/CD16(+) and CD56(bright)/CD16( ) NK cells were significantly decreased, and CD161-activating receptor expression by CD56(+)/CD3(+) NKT cells was significantly increased in women with implantation failures when compared with normal controls. CONCLUSIONS: An imbalance between inhibitory and activating receptor expression was found in NK cells of women with implantation failures. This imbalance may explain the adverse reproductive outcome. PMID- 15833100 TI - Immunological similarities between implantation and pre-eclampsia. AB - PROBLEM: Cytokines are involved in implantation success and failure. We envisage that they could be similarly involved in pre-eclampsia (PE). MATERIALS AND METHODS: First, we review the primipaternity and primiparity concepts and then why natural killer (NK) cells are involved in implantation. We stress that the common event in all PE is vascular remodelling. RESULTS AND CONCLUSION: We conclude that PE could involve cytokine and/or NK dysfunctions, and propose a working hypothesis. PMID- 15833101 TI - Nitric oxide levels and nitric oxide synthase expression in uterine samples from mares susceptible and resistant to persistent breeding-induced endometritis. AB - PROBLEM: Breeding-induced endometritis (BIE) in the mare is resolved by 36 hr after insemination in resistant mares. However, 10-15% susceptible broodmares fail to do so because of impaired uterine contractility between 7 and 19 hr after exposure to seminal or bacterial challenge, which reduces their fertility. METHOD OF STUDY: Nitric oxide (NO) in uterine secretions, and expression of nitric oxide synthase (NOS) in uterine biopsies were compared between susceptible and resistant groups 13 hr after insemination. RESULTS: Susceptible mares had a higher NO in their uterine secretions and greater inducible NOS (iNOS) expression in their biopsies compared with resistant mares. CONCLUSIONS: The NO mediates smooth muscle relaxation, but its role in persistent BIE has not been determined. Our data suggests a possible role of NO, either directly or in a NO-associated pathway, in delayed uterine clearance. PMID- 15833102 TI - Transient expression of CC chemokine TECK in the ovary during ovulation: its potential role in ovulation. AB - PROBLEM: Chemokine thymus-expressed chemokine (TECK), which is expressed exclusively in the thymus and small intestine, plays a critical role in T-cell development. Our previous study revealed its expression in the ovary also. This study investigated its ovarian expression during ovulatory process. METHOD OF STUDY: Super-ovulation was induced in young female CD1 mice by equine chorionic gonadotropin (eCG) and human chorionic gonadotropic (hCG). Ovarian TECK expression during ovulation was determined by: (1) reverse transcriptase polymerase chain reaction (RT-PCR) at mRNA level, (2) Western blot and immunohistology at the protein level, and (3) leukocyte infiltration assay at the bioactive level. RESULTS: A transient, high-level expression of TECK in murine ovaries at the mRNA level during hCG-induced ovulation was detected. Sequencing of directly cloned PCR product confirmed the ovarian expression of TECK. The peak expression of TECK was observed at 10-12 hr post-hCG injection; real-time PCR revealed an 800-fold increase during its expression peak over 0 hr. The expressed ovarian TECK protein was readily detectable by Western blot. Immunohistochemistry localized TECK expression to the ovarian interstitial tissue surrounding, or in the theca layer of the mature follicles undergoing ovulatory process. Expression of TECK receptor, the CC chemokine receptor (CCR9) was also detected in the ovulating ovaries. Using in vitro leukocyte infiltration assay, we first demonstrated that ovaries undergoing the ovulatory process were able to selectively chemoattract mononuclear cells. Importantly, neutralization of TECK by the antibody resulted in a 85% reduction in the chemotactic activities of the ovaries. CONCLUSION: This study suggested that ovarian expression of TECK is under a tight hormonal regulation, and expressed TECK may be responsible for recruitment of mononuclear cells into the ovary to participate in the ovulatory process. PMID- 15833103 TI - Expression of inhibitory-motif killer immunoglobulin-like receptor, KIR2DL1, is increased in natural killer cells from women with pelvic endometriosis. AB - PROBLEM: We investigated inhibitory and activation motif expression of killer immunoglobulin-like receptor (KIR) by natural killer (NK) cells, which may be pathogenetically involved in endometriosis. METHOD OF STUDY: We compared cells from 24 Japanese women laparoscopically diagnosed with endometriosis, to cells from 25 women with other laparoscopic diagnoses. KIR expression by NK cells was assessed in peripheral blood (PB) and peritoneal fluid (PF) by flow cytometry. Intracellular immunoreceptor tyrosine-based (IT) inhibitory and activation motifs (ITIM and ITAM) of KIR in PB was assessed by Western blotting. RESULTS: ITIM-KIR expression by PB NK cells was significantly and similarly greater than ITAM-KIR expression in women with and without endometriosis. Percentages of CD56(+) NK cells in PB and PF did not differ significantly between women with and without endometriosis; however, the percentage of CD158a(+) cells among CD56(+) NK cells in PB and PF was significantly higher in women with than without endometriosis. CONCLUSIONS: ITIM-KIR expressing NK cells might confer tolerance to peritoneal endometriotic implants. PMID- 15833104 TI - Effect of bacterial virulence on IL-18 expression in the amnion infected with Escherichia coli. AB - PROBLEM: The upregulation of inflammatory substances threatens pregnancy. Interleukin-18 (IL-18) is elevated in women who miscarried. The purpose of this study was to develop a pig model of chorioamnionitis to study the effect of bacterial virulence on IL-18 response in experimentally infected amnion. METHOD OF STUDY: A total of 20,000 colony-forming units of Escherichia coli (an enteropathogenic O55 strain, EPEC or O86 non-pathogenic strain) were administered into the amniotic cavity of pig fetuses at 70% of gestation for 10 hr. Fetal amniotic fluid samples were analyzed for IL-18 levels by enzyme-linked immunosorbent assay. The expression of IL-18 was studied also by immunohistochemistry on cryostat sections through amniotic membranes and pathological changes were observed by electron microscopy. RESULTS: Both E. coli strains propagated in amniotic fluids and reached similar counts. Only EPEC, however, caused a significant increase of IL-18 amniotic fluid levels (P < 0.001) and cytokine expression in the amniotic epithelium. CONCLUSIONS: The levels of IL 18 in infected amniotic fluids correlated with bacterial virulence and pathological changes in the amnion. PMID- 15833105 TI - Educational and labor wastage of doctors in Mexico: towards the construction of a common methodology. AB - BACKGROUND: This paper addresses the problem of wastage of the qualified labor force, which takes place both during the education process and when trained personnel try to find jobs in the local market. METHODS: Secondary sources were used, mainly the Statistical yearbooks of the National Association of Universities and Higher Education Institutions (ANUIES in Spanish). Also, the 2000 Population Census was used to estimate the different sources of labor market wastage. The formulas were modified to estimate educational and labor wastage rates. RESULTS: Out of every 1000 students who started a medical training in 1996, over 20% were not able to finish the training by 2000. Furthermore, out of every 1000 graduates, 31% were not able to find a remunerated position in the labor market that would enable them to put into practice the abilities and capacities obtained at school. Important differences can be observed between generalists and specialists, as well as between men and women. In the case of specialists and men, lower wastage rates can be observed as compared to the wastage rates of generalists and women. A large percentage of women dedicate themselves exclusively to household duties, which in labor terms represents a wastage of their capacity to participate in the production of formal health services. CONCLUSION: Women are becoming a majority in most medical schools, yet their participation in the labor market does not reflect the same trend. Among men, policies should be formulated to incorporate doctors in the specific health field for which they were trained. Regarding women, specific policies should target those who are dedicated full-time to household activities in order to create the possibility of having them occupy a remunerated job if they are willing to do so. Reducing wastage at both the educational and labor levels should improve the capacity of social investment, thereby increasing the capacity of the health system as a whole to provide services, particularly to those populations who are most in need. PMID- 15833106 TI - Inhibitors of mitogen-activated protein kinases differentially regulate costimulated T cell cytokine production and mouse airway eosinophilia. AB - BACKGROUND: T cells play a dominant role in the pathogenesis of asthma. Costimulation of T cells is necessary to fully activate them. An inducible costimulator (ICOS) of T cells is predominantly expressed on Th2 cells. Therefore, interference of signaling pathways precipitated by ICOS may present new therapeutic options for Th2 dominated diseases such as asthma. However, these signaling pathways are poorly characterized in vitro and in vivo. METHODS: Human primary CD4+ T cells from blood were activated by beads with defined combinations of surface receptor stimulating antibodies and costimulatory receptor ligands. Real-time RT-PCR was used for measuring the production of cytokines from activated T cells. Activation of mitogen activated protein kinase (MAPK) signaling pathways leading to cytokine synthesis were investigated by western blot analysis and by specific inhibitors. The effect of inhibitors in vivo was tested in a murine asthma model of late phase eosinophilia. Lung inflammation was assessed by differential cell count of the bronchoalveolar lavage, determination of serum IgE and lung histology. RESULTS: We showed in vitro that ICOS and CD28 are stimulatory members of an expanding family of co-receptors, whereas PD1 ligands failed to co-stimulate T cells. ICOS and CD28 activated different MAPK signaling cascades necessary for cytokine activation. By means of specific inhibitors we showed that p38 and ERK act downstream of CD28 and that ERK and JNK act downstream of ICOS leading to the induction of various T cell derived cytokines. Using a murine asthma model of late phase eosinophilia, we demonstrated that the ERK inhibitor U0126 and the JNK inhibitor SP600125 inhibited lung inflammation in vivo. This inhibition correlated with the inhibition of Th2 cytokines in the BAL fluid. Despite acting on different signaling cascades, we could not detect synergistic action of any combination of MAPK inhibitors. In contrast, we found that the p38 inhibitor SB203580 antagonizes the action of the ERK inhibitor U0126 in vitro and in vivo. CONCLUSION: These results demonstrate that the MAPKs ERK and JNK may be suitable targets for anti-inflammatory therapy of asthma, whereas inhibition of p38 seems to be an unlikely target. PMID- 15833107 TI - Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT PCR. AB - BACKGROUND: Preparation of RNA free from DNA is a critical step before performing RT-PCR assay. Total RNA isolated from several sources, including those obtained from Saccharomyces cerevisiae, using routine methodologies are frequently contaminated with DNA, which can give rise to amplification products that mimic the amplicons expected from the RNA target. RESULTS: We investigated the efficiency of two DNase I based protocols for eliminating DNA contaminations from RNA samples obtained from yeast cells. Both procedures are very efficient in eliminating DNA contamination from RNA samples and entail three main steps, which involve treating of RNA samples with DNase I, inhibition of the enzyme by EDTA and its subsequent inactivation at 65 degrees C. The DNase I treated samples were further purified with phenol: chloroform followed by precipitation with ice-cold ethanol (protocol I) or, alternatively, they were directly used in RT-PCR reactions (protocol II). Transcripts from ACT1, PDA1, CNA1, CNA2, TPS1 and TPS2 analyzed after each treatment showed that all mRNAs tested can be amplified if total RNA was extracted and purified after DNase I treatment, however, only TPS1, TPS2 and ACT1 mRNAs were amplified without extraction/purification step. CONCLUSION: Although more laborious and requiring a higher initial amount of material, the inclusion of an extraction and purification step allows to prepare RNA samples that are free from DNA and from low molecular contaminants and can be applied to amplify any Saccharomyces cerevisiae mRNA by RT-PCR. PMID- 15833108 TI - Cerebral infarction in diabetes: clinical pattern, stroke subtypes, and predictors of in-hospital mortality. AB - BACKGROUND: To compare the characteristics and prognostic features of ischemic stroke in patients with diabetes and without diabetes, and to determine the independent predictors of in-hospital mortality in people with diabetes and ischemic stroke. METHODS: Diabetes was diagnosed in 393 (21.3%) of 1,840 consecutive patients with cerebral infarction included in a prospective stroke registry over a 12-year period. Demographic characteristics, cardiovascular risk factors, clinical events, stroke subtypes, neuroimaging data, and outcome in ischemic stroke patients with and without diabetes were compared. Predictors of in-hospital mortality in diabetic patients with ischemic stroke were assessed by multivariate analysis. RESULTS: People with diabetes compared to people without diabetes presented more frequently atherothrombotic stroke (41.2% vs 27%) and lacunar infarction (35.1% vs 23.9%) (P < 0.01). The in-hospital mortality in ischemic stroke patients with diabetes was 12.5% and 14.6% in those without (P = NS). Ischemic heart disease, hyperlipidemia, subacute onset, 85 years old or more, atherothrombotic and lacunar infarcts, and thalamic topography were independently associated with ischemic stroke in patients with diabetes, whereas predictors of in-hospital mortality included the patient's age, decreased consciousness, chronic nephropathy, congestive heart failure and atrial fibrillation CONCLUSION: Ischemic stroke in people with diabetes showed a different clinical pattern from those without diabetes, with atherothrombotic stroke and lacunar infarcts being more frequent. Clinical factors indicative of the severity of ischemic stroke available at onset have a predominant influence upon in-hospital mortality and may help clinicians to assess prognosis more accurately. PMID- 15833109 TI - Platelet-activating factor enhancement of calcium influx and interleukin-6 expression, but not production, in human microglia. AB - Calcium-sensitive fluorescence microscopy and molecular biology analysis have been used to study the effects of platelet-activating factor (PAF) on intracellular calcium [Ca2+]i and IL-6 expression in human microglia. PAF (applied acutely at 100 nM) elicited a biphasic response in [Ca2+]i consisting of an initial rapid increase of [Ca2+]i due to release from internal stores, followed by a sustained influx. The latter phase of the [Ca2+]i increase was blocked by SKF96365, a non-selective store-operated channel (SOC) inhibitor. RT PCR analysis showed PAF treatment of microglia induced expression of the pro inflammatory cytokine IL-6 in a time-dependent manner which was blocked in the presence of SKF96365. However, ELISA assay showed no production of IL-6 was elicited at any time point (1-24 h) for microglial exposures to PAF. These findings suggest that PAF stimulation of human microglia induces expression, but not production, of IL-6 and that SOC-mediated [Ca2+]i influx contributes to the enhanced expression of the cytokine. PMID- 15833110 TI - Skin testing versus radioallergosorbent testing for indoor allergens. AB - BACKGROUND: Skin testing (ST) is the most common screening method for allergy evaluation. Measurement of serum specific IgE is also commonly used, but less so by allergists than by other practitioners. The sensitivity and specificity of these testing methods may vary by type of causative allergen and type of allergic manifestation. We compared ST reactivity with serum specific IgE antibodies to common indoor allergens in patients with respiratory allergies. METHODS: 118 patients (3 mo-58 yr, mean 12 yr) with allergic rhinitis and/or bronchial asthma had percutaneous skin testing (PST) supplemented by intradermal testing (ID) with those allergens suspected by history but showed negative PST. The sera were tested blindly for specific IgE antibodies by the radioallergosorbent test (Phadebas RAST). The allergens were D. farinae (118), cockroach (60), cat epithelium (90), and dog epidermal (90). Test results were scored 0-4; ST >/= 2 + and RAST >/= 1 + were considered positive. RESULTS: The two tests were in agreement (i.e., either both positive or both negative) in 52.2% (dog epidermal) to 62.2% (cat epithelium). When RAST was positive, ST was positive in 80% (dog epidermal) to 100% (cockroach mix). When ST was positive, RAST was positive in 16.3% (dog epidermal) to 50.0% (D. farinae). When RAST was negative, ST was positive in 48.5% (cat epithelium) to 69.6% (D. farinae). When ST was negative, RAST was positive in 0% (cockroach) to 5.6% (cat epithelium). The scores of ST and RAST showed weak to moderate correlation (r = 0.24 to 0.54). Regardless of history of symptoms on exposure, ST was superior to RAST in detecting sensitization to cat epithelium and dog epidermal. CONCLUSION: For all four indoor allergens tested, ST was more sensitive than RAST. When both tests were positive, their scores showed poor correlation. Sensitizations to cat epithelium and dog epidermal are common, even in subjects who claimed no direct exposure. PMID- 15833111 TI - Comparable response of ccn1 with ccn2 genes upon arthritis: An in vitro evaluation with a human chondrocytic cell line stimulated by a set of cytokines. AB - BACKGROUND: The chondrosarcoma-derived HCS-2/8 has been known to be an excellent model of human articular chondrocytes. By mimicking the arthritic conditions through the treatment of HCS-2/8 cells with cytokines, we estimated the gene expression response of ccn1 and ccn2 during the course of joint inflammation in vitro. RESULTS: In order to mimic the initiation of inflammation, HCS-2/8 cells were treated with tumor necrosis factor (TNF)-alpha. To induce pro-inflammatory or reparative responses, TGF-beta was employed. Effects of an anti-inflammatory glucocorticoid were also evaluated. After stimulation, expression levels of ccn1 and ccn2 were quantitatively analyzed. Surprisingly, not only ccn2, but also ccn1 expression was repressed upon TNF-alpha stimulation, whereas both mRNAs were uniformly induced by transforming growth factor (TGF)-beta and a glucocorticoid. CONCLUSION: These results describing the same response during the course of inflammation suggest similar and co-operative roles of these 2 ccn family members in the course of arthritis. PMID- 15833114 TI - Life after 45 and before 60: the Retrovirology Prize. AB - Retrovirology announces an annual prize to recognize one outstanding mid-career retrovirologist. PMID- 15833113 TI - Molecular advances in the cell biology of SARS-CoV and current disease prevention strategies. AB - In the aftermath of the SARS epidemic, there has been significant progress in understanding the molecular and cell biology of SARS-CoV. Some of the milestones are the availability of viral genome sequence, identification of the viral receptor, development of an infectious cDNA clone, and the identification of viral antigens that elicit neutralizing antibodies. However, there is still a large gap in our understanding of how SARS-CoV interacts with the host cell and the rapidly changing viral genome adds another variable to this equation. Now the SARS-CoV story has entered a new phase, a search for preventive strategies and a cure for the disease. This review highlights the progress made in identifying molecular aspects of SARS-CoV biology that is relevant in developing disease prevention strategies. Authors conclude that development of successful SARS-CoV vaccines and antivirals depends on the progress we make in these areas in the immediate future. PMID- 15833115 TI - Barriers to coliphage infection of commensal intestinal flora of laboratory mice. AB - BACKGROUND: Growth characteristics of coliphage viruses indicate that they are adapted to live with their Eschericia coli hosts in the intestinal tract. However, coliphage experimentally introduced by ingestion persist only transiently if at all in the gut of humans and other animals. This study attempted to identify the barriers to long term establishment of exogenous coliphage in the gastrointestinal (GI) tracts of laboratory mice. Intestinal contents were screened for the presence of coliphage and host bacteria, and strains of E. coli bacteria from different segments of the GI tract were tested for susceptibility to six common laboratory coliphages. RESULTS: Contrary to expectations, coliphage were not evident in the GI tracts of laboratory mice, although they were occasionally detected in feces. Commensal flora showed extreme variability within groups of mice despite identical handling and diet. Less than 20% of 48 mice tested carried E. coli in their gut, and of 22 commensal E. coli strains isolated and tested, 59% were completely resistant to infection by lambda, M13, P1, T4, T7, and PhiX174 coliphage. Lysogeny could not be demonstrated in the commensal strains as mitomycin C failed to induce detectable phage. Pre-existing immunity to phages was not evident as sera and fecal washes did not contain significant antibody titers to six laboratory phage types. CONCLUSION: Lack of sufficient susceptible host bacteria seems to be the most likely barrier to establishment of new coliphage infections in the mouse gut. PMID- 15833116 TI - Use of the femoral vein ('groin injecting') by a sample of needle exchange clients in Bristol, UK. AB - BACKGROUND: Use of the femoral vein for intravenous access by injecting drug users (IDUs) (commonly called 'groin injecting') is a practice that is often observed but on which little is written in the literature. The purpose of this study was to describe self-reported data from a sample of groin injectors on the natural history and rationale regarding their groin injecting, to inform future research and the development of appropriate harm reduction strategies. METHODS: A convenience sample of groin injectors willing to participate in a semi-structured interview were recruited through the Bristol Drugs Project Harm Reduction Service. The interviews were conducted over the period of one week. Data on transition to groin injecting, rationale for use and incidence of problems were collected. RESULTS: Forty seven IDUs currently injecting in their femoral vein ('groin') were interviewed, 66% (n = 31) male and 34% (n = 16) female. Their mean age was 31 yrs (range 17 to 50 yrs; SD = 7.7). The mean length of time since first injecting episode was 9.6 yrs (range 6 mths to 30 yrs; SD = 7.0). The mean length of time since use of the groin began was 2.6 years (range 1 mth to 15 yrs; SD = 3.3). The mean length of time between first injection and first use of the groin was 7.0 yrs (SD = 7.0). One person had used no other area for venous access prior to using the groin, nine people had used one, nine people had used two, 10 people had used three, five people had used four and 13 people had used more than four areas. The main reason given for starting to inject in the groin was that 'no other sites were left'. However further discussion identified this meant no other convenient sites were accessible. Practises such as the rotation of injecting sites, as advocated in many harm reduction leaflets, were reported to be difficult and unreliable. The risk of missing the vein and subsequently losing the 'hit' was considered high. Use of the non-dominant hand to administer injections was problematic and deterred rotation between arms. The groin site was reported to be convenient, provide quick access, with little mess and less pain than smaller more awkward veins. The formation of sinuses over time facilitated continued use of the groin. Approximately two thirds of participants had experienced difficulty gaining IV access at their groin. Common problem included scar tissue occlusion, swelling and pain. Some reported infections and past history of deep vein thrombosis. CONCLUSION: Use of the groin was perceived to be convenient by the study group. Problems following safer injecting advice were identified, including dexterity difficulties leading to fear of losing the 'hit'. Health problems at the groin site did not deter use. These results suggest further qualitative work is needed to explore the difficulties in following safer injecting advice in more detail and inform the development of more appropriate advice. Further quantitative work is necessary to establish the prevalence of groin injecting amongst IDUs and the incidence of associated problems. There is a need for a longitudinal study to examine the relationship between injecting technique and loss of patency of veins. If protective factors could be identified, evidence-based safer injecting advice could be established to preserve peripheral veins and reduce use of the groin site, which is high risk and associated with serious adverse consequences. PMID- 15833117 TI - Genome-wide prediction and identification of cis-natural antisense transcripts in Arabidopsis thaliana. AB - BACKGROUND: Natural antisense transcripts (NAT) are a class of endogenous coding or non-protein-coding RNAs with sequence complementarity to other transcripts. Several lines of evidence have shown that cis- and trans-NATs may participate in a broad range of gene regulatory events. Genome-wide identification of cis-NATs in human, mouse and rice has revealed their widespread occurrence in eukaryotes. However, little is known about cis-NATs in the model plant Arabidopsis thaliana. RESULTS: We developed a new computational method to predict and identify cis encoded NATs in Arabidopsis and found 1,340 potential NAT pairs. The expression of both sense and antisense transcripts of 957 NAT pairs was confirmed using Arabidopsis full-length cDNAs and public massively parallel signature sequencing (MPSS) data. Three known or putative Arabidopsis imprinted genes have cis antisense transcripts. Sequences and the genomic arrangement of two Arabidopsis NAT pairs are conserved in rice. CONCLUSION: We combined information from full length cDNAs and Arabidopsis genome annotation in our NAT prediction work and reported cis-NAT pairs that could not otherwise be identified by using one of the two datasets only. Analysis of MPSS data suggested that for most Arabidopsis cis NAT pairs, there is predominant expression of one of the two transcripts in a tissue-specific manner. PMID- 15833118 TI - Conservation of tandem stop codons in yeasts. AB - BACKGROUND: It has been long thought that the stop codon in a gene is followed by another stop codon that acts as a backup if the real one is read through by a near-cognate tRNA. The existence of such 'tandem stop codons', however, remains elusive. RESULTS: Here we show that a statistical excess of stop codons has evolved at the third codon downstream of the real stop codon UAA in yeasts. Comparative analysis indicates that stop codons at this location are considerably more conserved than sense codons, suggesting that these tandem stop codons are maintained by selection. We evaluated the influence of expression levels of genes and other biological factors on the distribution of tandem stop codons. Our results suggest that expression level is an important factor influencing the presence of tandem stop codons. CONCLUSIONS: Our study demonstrates the existence of tandem stop codons, which represent one of many meaningful genomic features that are driven by relatively weak selective forces. PMID- 15833119 TI - Contribution of transcriptional regulation to natural variations in Arabidopsis. AB - BACKGROUND: Genetic control of gene transcription is a key component in genome evolution. To understand the transcriptional basis of natural variation, we have studied genome-wide variations in transcription and characterized the genetic variations in regulatory elements among Arabidopsis accessions. RESULTS: Among five accessions (Col-0, C24, Ler, WS-2, and NO-0) 7,508 probe sets with no detectable genomic sequence variations were identified on the basis of the comparative genomic hybridization to the Arabidopsis GeneChip microarray, and used for accession-specific transcriptome analysis. Two-way ANOVA analysis has identified 60 genes whose mRNA levels differed in different accession backgrounds in an organ-dependent manner. Most of these genes were involved in stress responses and late stages of plant development, such as seed development. Correlation analysis of expression patterns of these 7,508 genes between pairs of accessions identified a group of 65 highly plastic genes with distinct expression patterns in each accession. CONCLUSION: Genes that show substantial genetic variation in mRNA level are those with functions in signal transduction, transcription and stress response, suggesting the existence of variations in the regulatory mechanisms for these genes among different accessions. This is in contrast to those genes with significant polymorphisms in the coding regions identified by genomic hybridization, which include genes encoding transposon related proteins, kinases and disease-resistance proteins. While relatively fewer sequence variations were detected on average in the coding regions of these genes, a number of differences were identified from the upstream regions, several of which alter potential cis-regulatory elements. Our results suggest that nucleotide polymorphisms in regulatory elements of genes encoding controlling factors could be primary targets of natural selection and a driving force behind the evolution of Arabidopsis accessions. PMID- 15833120 TI - Promoter features related to tissue specificity as measured by Shannon entropy. AB - BACKGROUND: The regulatory mechanisms underlying tissue specificity are a crucial part of the development and maintenance of multicellular organisms. A genome-wide analysis of promoters in the context of gene-expression patterns in tissue surveys provides a means of identifying the general principles for these mechanisms. RESULTS: We introduce a definition of tissue specificity based on Shannon entropy to rank human genes according to their overall tissue specificity and by their specificity to particular tissues. We apply our definition to microarray-based and expressed sequence tag (EST)-based expression data for human genes and use similar data for mouse genes to validate our results. We show that most genes show statistically significant tissue-dependent variations in expression level. We find that the most tissue-specific genes typically have a TATA box, no CpG island, and often code for extracellular proteins. As expected, CpG islands are found in most of the least tissue-specific genes, which often code for proteins located in the nucleus or mitochondrion. The class of genes with no CpG island or TATA box are the most common mid-specificity genes and commonly code for proteins located in a membrane. Sp1 was found to be a weak indicator of less-specific expression. YY1 binding sites, either as initiators or as downstream sites, were strongly associated with the least-specific genes. CONCLUSIONS: We have begun to understand the components of promoters that distinguish tissue-specific from ubiquitous genes, to identify associations that can predict the broad class of gene expression from sequence data alone. PMID- 15833121 TI - A genomic approach to investigate developmental cell death in woody tissues of Populus trees. AB - BACKGROUND: Poplar (Populus sp.) has emerged as the main model system for molecular and genetic studies of forest trees. A Populus expressed sequence tag (EST) database (POPULUSDB) was previously created from 19 cDNA libraries each originating from different Populus tree tissues, and opened to the public in September 2004. We used this dataset for in silico transcript profiling of a particular process in the woody tissues of the Populus stem: the programmed death of xylem fibers. RESULTS: One EST library in POPULUSDB originates from woody tissues of the Populus stem where xylem fibers undergo cell death. Analysis of EST abundances and library distribution within the POPULUSDB revealed a large number of previously uncharacterized transcripts that were unique in this library and possibly related to the death of xylem fibers. The in silico analysis was complemented by a microarray analysis utilizing a novel Populus cDNA array with a unigene set of 25,000 sequences. CONCLUSIONS: In silico analysis, combined with the microarray analysis, revealed the usefulness of non-normalized EST libraries in elucidating transcriptional regulation of previously uncharacterized physiological processes. The data suggested the involvement of two novel extracellular serine proteases, nodulin-like proteins and an Arabidopsis thaliana OPEN STOMATA 1 (AtOST1) homolog in signaling fiber-cell death, as well as mechanisms responsible for hormonal control, nutrient remobilization, regulation of vacuolar integrity and autolysis of the dying fibers. PMID- 15833122 TI - An evolutionary and functional assessment of regulatory network motifs. AB - BACKGROUND: Cellular functions are regulated by complex webs of interactions that might be schematically represented as networks. Two major examples are transcriptional regulatory networks, describing the interactions among transcription factors and their targets, and protein-protein interaction networks. Some patterns, dubbed motifs, have been found to be statistically over represented when biological networks are compared to randomized versions thereof. Their function in vitro has been analyzed both experimentally and theoretically, but their functional role in vivo, that is, within the full network, and the resulting evolutionary pressures remain largely to be examined. RESULTS: We investigated an integrated network of the yeast Saccharomyces cerevisiae comprising transcriptional and protein-protein interaction data. A comparative analysis was performed with respect to Candida glabrata, Kluyveromyces lactis, Debaryomyces hansenii and Yarrowia lipolytica, which belong to the same class of hemiascomycetes as S. cerevisiae but span a broad evolutionary range. Phylogenetic profiles of genes within different forms of the motifs show that they are not subject to any particular evolutionary pressure to preserve the corresponding interaction patterns. The functional role in vivo of the motifs was examined for those instances where enough biological information is available. In each case, the regulatory processes for the biological function under consideration were found to hinge on post-transcriptional regulatory mechanisms, rather than on the transcriptional regulation by network motifs. CONCLUSION: The overabundance of the network motifs does not have any immediate functional or evolutionary counterpart. A likely reason is that motifs within the networks are not isolated, that is, they strongly aggregate and have important edge and/or node sharing with the rest of the network. PMID- 15833123 TI - Quantitative genomics of starvation stress resistance in Drosophila. AB - BACKGROUND: A major challenge of modern biology is to understand the networks of interacting genes regulating complex traits, and the subset of these genes that affect naturally occurring quantitative genetic variation. Previously, we used P element mutagenesis and quantitative trait locus (QTL) mapping in Drosophila to identify candidate genes affecting resistance to starvation stress, and variation in resistance to starvation stress between the Oregon-R (Ore) and 2b strains. Here, we tested the efficacy of whole-genome transcriptional profiling for identifying genes affecting starvation stress resistance. RESULTS: We evaluated whole-genome transcript abundance for males and females of Ore, 2b, and four recombinant inbred lines derived from them, under control and starved conditions. There were significant differences in transcript abundance between the sexes for nearly 50% of the genome, while the transcriptional response to starvation stress involved approximately 25% of the genome. Nearly 50% of P-element insertions in 160 genes with altered transcript abundance during starvation stress had mutational effects on starvation tolerance. Approximately 5% of the genome exhibited genetic variation in transcript abundance, which was largely attributable to regulation by unlinked genes. Genes exhibiting variation in transcript abundance among lines did not cluster within starvation resistance QTLs, and none of the candidate genes affecting variation in starvation resistance between Ore and 2b exhibited significant differences in transcript abundance between lines. CONCLUSIONS: Expression profiling is a powerful method for identifying networks of pleiotropic genes regulating complex traits, but the relationship between variation in transcript abundance among lines used to map QTLs and genes affecting variation in quantitative traits is complicated. PMID- 15833124 TI - The microbial selenoproteome of the Sargasso Sea. AB - BACKGROUND: Selenocysteine (Sec) is a rare amino acid which occurs in proteins in major domains of life. It is encoded by TGA, which also serves as the signal for termination of translation, precluding identification of selenoprotein genes by available annotation tools. Information on full sets of selenoproteins (selenoproteomes) is essential for understanding the biology of selenium. Herein, we characterized the selenoproteome of the largest microbial sequence dataset, the Sargasso Sea environmental genome project. RESULTS: We identified 310 selenoprotein genes that clustered into 25 families, including 101 new selenoprotein genes that belonged to 15 families. Most of these proteins were predicted redox proteins containing catalytic selenocysteines. Several bacterial selenoproteins previously thought to be restricted to eukaryotes were detected by analyzing eukaryotic and bacterial SECIS elements, suggesting that eukaryotic and bacterial selenoprotein sets partially overlapped. The Sargasso Sea microbial selenoproteome was rich in selenoproteins and its composition was different from that observed in the combined set of completely sequenced genomes, suggesting that these genomes do not accurately represent the microbial selenoproteome. Most detected selenoproteins occurred sporadically compared to the widespread presence of their cysteine homologs, suggesting that many selenoproteins recently evolved from cysteine-containing homologs. CONCLUSIONS: This study yielded the largest selenoprotein dataset to date, doubled the number of prokaryotic selenoprotein families and provided insights into forces that drive selenocysteine evolution. PMID- 15833125 TI - Derivation of genetic interaction networks from quantitative phenotype data. AB - We have generalized the derivation of genetic-interaction networks from quantitative phenotype data. Familiar and unfamiliar modes of genetic interaction were identified and defined. A network was derived from agar-invasion phenotypes of mutant yeast. Mutations showed specific modes of genetic interaction with specific biological processes. Mutations formed cliques of significant mutual information in their large-scale patterns of genetic interaction. These local and global interaction patterns reflect the effects of gene perturbations on biological processes and pathways. PMID- 15833127 TI - Who owns the data? AB - About ten years ago, a group of scientists began to argue that it was unfair to ask other scientists to pay to read the results of research that had been publicly funded. PMID- 15833126 TI - Pilot Anopheles gambiae full-length cDNA study: sequencing and initial characterization of 35,575 clones. AB - We describe the preliminary analysis of over 35,000 clones from a full-length enriched cDNA library from the malaria mosquito vector Anopheles gambiae. The clones define nearly 3,700 genes, of which around 2,600 significantly improve current gene definitions. An additional 17% of the genes were not previously annotated, suggesting that an equal percentage may be missing from the current Anopheles genome annotation. PMID- 15833128 TI - Anatomical ontologies: names and places in biology. AB - Ontology has long been the preserve of philosophers and logicians. Recently, ideas from this field have been picked up by computer scientists as a basis for encoding knowledge and with the hope of achieving interoperability and intelligent system behavior. In bioinformatics, ontologies might allow hitherto impossible query and data-mining activities. We review the use of anatomy ontologies to represent space in biological organisms, specifically mouse and human. PMID- 15833129 TI - A non-random walk through the genome. AB - Recent publications on a wide range of eukaryotes indicate that genes showing particular expression patterns are not randomly distributed in the genome but are clustered into contiguous regions that we call neighborhoods. It seems probable that this organization is related to chromatin and the structure of the nucleus. PMID- 15833130 TI - Genomic studies of mood disorders -- the brain as a muscle? AB - Recent genomic studies showing abnormalities in the fibroblast growth factor system in the postmortem brains of people with major depressive disorder support previous indications of a role for growth factors in mood disorders. Similar molecular pathways, volumetric changes, and the effects of exercise on mood suggest a superficial analogy, and perhaps a deeper relationship, between muscle and brain functioning. PMID- 15833131 TI - SINEs point to abundant editing in the human genome. AB - Recent bioinformatic analyses suggest that almost all human transcripts are edited by adenosine deaminases (ADARs), converting adenosines to inosines. Most of this editing is in Alu element transcripts, which are unique to primates. This editing might have no function or might be involved in functions such as the regulation of splicing, chromatin or nuclear localization of transcripts. PMID- 15833133 TI - Diverse roles for RNA in gene regulation. PMID- 15833132 TI - Assessing the origin of species in the genomic era. AB - Advances in genomics have rapidly accelerated research into the genetics of species differences, reproductive isolating barriers, and hybrid incompatibility. Recent genomic analyses in Drosophila species suggest that modified olfactory cues are involved in discrimination that is reinforced by natural selection. PMID- 15833134 TI - Protein degradation, signaling, microRNAs and cancer. PMID- 15833135 TI - Making systems biology work in the 21st century. PMID- 15833136 TI - Rare mutations predisposing to familial adenomatous polyposis in Greek FAP patients. AB - BACKGROUND: Familial Adenomatous Polyposis (FAP) is caused by germline mutations in the APC (Adenomatous Polyposis Coli) gene. The vast majority of APC mutations are point mutations or small insertions/deletions which lead to truncated protein products. Splicing mutations or gross genomic rearrangements are less common inactivating events of the APC gene. METHODS: In the current study genomic DNA or RNA from ten unrelated FAP suspected patients was examined for germline mutations in the APC gene. Family history and phenotype were used in order to select the patients. Methods used for testing were dHPLC (denaturing High Performance Liquid Chromatography), sequencing, MLPA (Multiplex Ligation - dependent Probe Amplification), Karyotyping, FISH (Fluorescence In Situ Hybridization) and RT-PCR (Reverse Transcription - Polymerase Chain Reaction). RESULTS: A 250 Kbp deletion in the APC gene starting from intron 5 and extending beyond exon 15 was identified in one patient. A substitution of the +5 conserved nucleotide at the splice donor site of intron 9 in the APC gene was shown to produce frameshift and inefficient exon skipping in a second patient. Four frameshift mutations (1577insT, 1973delAG, 3180delAAAA, 3212delA) and a nonsense mutation (C1690T) were identified in the rest of the patients. CONCLUSION: Screening for APC mutations in FAP patients should include testing for splicing defects and gross genomic alterations. PMID- 15833137 TI - Transcriptome analysis in primary neural stem cells using a tag cDNA amplification method. AB - BACKGROUND: Neural stem cells (NSCs) can be isolated from the adult mammalian brain and expanded in culture, in the form of cellular aggregates called neurospheres. Neurospheres provide an in vitro model for studying NSC behaviour and give information on the factors and mechanisms that govern their proliferation and differentiation. They are also a promising source for cell replacement therapies of the central nervous system. Neurospheres are complex structures consisting of several cell types of varying degrees of differentiation. One way of characterising neurospheres is to analyse their gene expression profiles. The value of such studies is however uncertain since they are heterogeneous structures and different populations of neurospheres may vary significantly in their gene expression. RESULTS: To address this issue, we have used cDNA microarrays and a recently reported tag cDNA amplification method to analyse the gene expression profiles of neurospheres originating from separate isolations of the lateral ventricle wall of adult mice and passaged to varying degrees. Separate isolations as well as consecutive passages yield a high variability in gene expression while parallel cultures yield the lowest variability. CONCLUSIONS: We demonstrate a low technical amplification variability using the employed amplification strategy and conclude that neurospheres from the same isolation and passage are sufficiently similar to be used for comparative gene expression analysis. PMID- 15833138 TI - Development and validation of the Chinese Quality of Life Instrument. AB - BACKGROUND: This paper describes the development of the Chinese Quality of Life Instrument (ChQOL) which is a self-report health status instrument. Chinese Medicine relies very much on asking subjective feelings of patients in the process of diagnosis and monitoring of treatment. For thousands of years, Chinese Medicine practitioners have accumulated a good wealth of experiences in asking questions about health of their patients based on the concept of health in Chinese Medicine. These experiences were then transformed into questions for the ChQOL. It is believed that ChQOL can contribute to the existing Patient Report Outcome measures. This paper outlines the concept of health and disease in Traditional Chinese Medicine, the building of the conceptual framework of the ChQOL, the steps of drafting, selecting and validating the items, and the psychometric properties of the ChQOL. METHODS: The development of the ChQOL was based on the concept of health in Traditional Chinese Medicine with a theory driven approach. Based on the results of literature review, the research team developed an initial model of health which encompassed the concept of health in TCM. An expert panel was then invited to comment and give suggestions for improvement of the initial model. According to their suggestions, the model was refined and a set of initial items for the ChQOL was drafted. The refined model, together with the key domains, facets and initial items of the ChQOL were then mailed to a sample of about 100 Chinese medicine practitioners throughout Mainland China for their comments and advice. A revised set of items were developed for linguistic testing by a convenience sample consisting of both healthy people and people who attended Chinese Medicine treatment. After that, an item pool was developed for field-testing. Field test was conducted on a convenience sample of healthy and patient subjects to determine the construct validity and psychometric properties of the ChQOL. RESULTS: Construct validity was established by various methods, i.e. the internal consistency in all facets and domains were good; the correlation between facets to domain, and domains to overall ChQOL correlation were high; confirmatory factor analysis showed that the structure fitness of all facets, domain and overall structure were good with CFI > 0.9. Test-retest reliability was also good, especially in the domain scores with ICC value ranging from 0.83 to 0.90. No ceiling or floor effect was noted which indicated that ChQOL can be applied to subjects with a wide range of health status. Most facet scores, domain scores and the overall CHQOL scores were able to discriminate groups of subjects with known differences in health status. The ChQOL had mild positive convergence with the other generic health related QOL measures, i.e. the WHOQOL-100 and the SF-36, with moderate correlations. CONCLUSION: In conclusion, the study indicated that the ChQOL is conceptually valid with satisfactory psychometric properties. It can provide additional information on health and QOL on top of the existing generic health related QOL measures. Furthermore, it forms basis for further testing and applications in clinical trials. PMID- 15833140 TI - Predictors of glycemic control among patients with Type 2 diabetes: a longitudinal study. AB - BACKGROUND: Diabetes is the sixth leading cause of death and results in significant morbidity. The purpose of this study is to determine what demographic, health status, treatment, access/quality of care, and behavioral factors are associated with poor glycemic control in a Type 2 diabetic, low income, minority, San Diego population. METHODS: Longitudinal observational data was collected on patients with Type 2 diabetes from Project Dulce, a program in San Diego County designed to care for an underserved diabetic population. The study sample included 573 patients with a racial/ethnic mix of 53% Hispanic, 7% black, 18% Asian, 20% white, and 2% other. We utilized mixed effects models to determine the factors associated with poor glycemic control using hemoglobin A1C (A1C) as the outcome of interest. A multi-step model building process was used resulting in a final parsimonious model with main effects and interaction terms. RESULTS: Patients had a mean age of 55 years, 69% were female, the mean duration of diabetes was 7.1 years, 31% were treated with insulin, and 57% were obese. American Diabetes Association (ADA) recommendations for blood pressure and total cholesterol were met by 71% and 68%, respectively. Results of the mixed effects model showed that patients who were uninsured, had diabetes for a longer period of time, used insulin or multiple oral agents, or had high cholesterol had higher A1C values over time indicating poorer glycemic control. The younger subjects also had poorer control. CONCLUSION: This study provides factors that predict glycemic control in a specific low-income, multiethnic, Type 2 diabetic population. With this information, subgroups with high risk of disease morbidity were identified. Barriers that prevent these patients from meeting their goals must be explored to improve health outcomes. PMID- 15833139 TI - Microarray karyotyping of commercial wine yeast strains reveals shared, as well as unique, genomic signatures. AB - BACKGROUND: Genetic differences between yeast strains used in wine-making may account for some of the variation seen in their fermentation properties and may also produce differing sensory characteristics in the final wine product itself. To investigate this, we have determined genomic differences among several Saccharomyces cerevisiae wine strains by using a "microarray karyotyping" (also known as "array-CGH" or "aCGH") technique. RESULTS: We have studied four commonly used commercial wine yeast strains, assaying three independent isolates from each strain. All four wine strains showed common differences with respect to the laboratory S. cerevisiae strain S288C, some of which may be specific to commercial wine yeasts. We observed very little intra-strain variation; i.e., the genomic karyotypes of different commercial isolates of the same strain looked very similar, although an exception to this was seen among the Montrachet isolates. A moderate amount of inter-strain genomic variation between the four wine strains was observed, mostly in the form of depletions or amplifications of single genes; these differences allowed unique identification of each strain. Many of the inter-strain differences appear to be in transporter genes, especially hexose transporters (HXT genes), metal ion sensors/transporters (CUP1, ZRT1, ENA genes), members of the major facilitator superfamily, and in genes involved in drug response (PDR3, SNQ1, QDR1, RDS1, AYT1, YAR068W). We therefore used halo assays to investigate the response of these strains to three different fungicidal drugs (cycloheximide, clotrimazole, sulfomethuron methyl). Strains with fewer copies of the CUP1 loci showed hypersensitivity to sulfomethuron methyl. CONCLUSION: Microarray karyotyping is a useful tool for analyzing the genome structures of wine yeasts. Despite only small to moderate variations in gene copy numbers between different wine yeast strains and within different isolates of a given strain, there was enough variation to allow unique identification of strains; additionally, some of the variation correlated with drug sensitivity. The relatively small number of differences seen by microarray karyotyping between the strains suggests that the differences in fermentative and organoleptic properties ascribed to these different strains may arise from a small number of genetic changes, making it possible to test whether the observed differences do indeed confer different sensory properties in the finished wine. PMID- 15833141 TI - Altered expression of membrane-bound and soluble CD95/Fas contributes to the resistance of fibrotic lung fibroblasts to FasL induced apoptosis. AB - BACKGROUND: An altered susceptibility of lung fibroblasts to Fas-induced apoptosis has been implicated in the pathogenesis of pulmonary fibrosis; however, the underlying mechanism is not completely understood. Here, we studied the susceptibility of lung fibroblasts, obtained from patients with (f-fibs) and without pulmonary fibrosis (n-fibs), to FasL- (CD95L/APO-1) induced apoptosis in relation to the expression and the amounts of membrane-bound and soluble Fas. We also analysed the effects of tumor necrosis factor-beta on FasL-induced cell death. METHODS: Apoptosis was induced with recombinant human FasL, with and without prior stimulation of the fibroblasts with tumor necrosis factor-alpha and measured by a histone fragmentation assay and flow cytometry. The expression of Fas mRNA was determined by quantitative PCR. The expression of cell surface Fas was determined by flow cytometry, and that of soluble Fas (sFas) was determined by enzyme-linked immunosorbent assay. RESULTS: When compared to n-fibs, f-fibs were resistant to FasL-induced apoptosis, despite significantly higher levels of Fas mRNA. F-fibs showed lower expression of surface-bound Fas but higher levels of sFas. While TNF-alpha increased the susceptibility to FasL-induced apoptosis in n-fibs, it had no pro-apoptotic effect in f-fibs. CONCLUSIONS: The data suggest that lower expression of surface Fas, but higher levels of apoptosis inhibiting sFas, contribute to the resistance of fibroblasts in lung fibrosis against apoptosis, to increased cellularity and also to increased formation and deposition of extracellular matrix. PMID- 15833142 TI - Filtering high-throughput protein-protein interaction data using a combination of genomic features. AB - BACKGROUND: Protein-protein interaction data used in the creation or prediction of molecular networks is usually obtained from large scale or high-throughput experiments. This experimental data is liable to contain a large number of spurious interactions. Hence, there is a need to validate the interactions and filter out the incorrect data before using them in prediction studies. RESULTS: In this study, we use a combination of 3 genomic features -- structurally known interacting Pfam domains, Gene Ontology annotations and sequence homology -- as a means to assign reliability to the protein-protein interactions in Saccharomyces cerevisiae determined by high-throughput experiments. Using Bayesian network approaches, we show that protein-protein interactions from high-throughput data supported by one or more genomic features have a higher likelihood ratio and hence are more likely to be real interactions. Our method has a high sensitivity (90%) and good specificity (63%). We show that 56% of the interactions from high throughput experiments in Saccharomyces cerevisiae have high reliability. We use the method to estimate the number of true interactions in the high-throughput protein-protein interaction data sets in Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens to be 27%, 18% and 68% respectively. Our results are available for searching and downloading at http://helix.protein.osaka u.ac.jp/htp/. CONCLUSION: A combination of genomic features that include sequence, structure and annotation information is a good predictor of true interactions in large and noisy high-throughput data sets. The method has a very high sensitivity and good specificity and can be used to assign a likelihood ratio, corresponding to the reliability, to each interaction. PMID- 15833144 TI - Modulating co-stimulation: a rational strategy in the treatment of rheumatoid arthritis? AB - Rheumatoid arthritis (RA) is a common destructive inflammatory disease that affects 0.5-1% of the population in many countries. Even though several new treatments have been introduced for patients with RA, a considerable proportion of patients do not benefit from these, and the need for alternative treatment strategies is clear. This review explores the potential for a therapy targeting the adaptive immune system by modulating co-stimulation of T cells with a CTLA4 Ig fusion protein (abatacept). PMID- 15833145 TI - The evolving clinical profile of abatacept (CTLA4-Ig): a novel co-stimulatory modulator for the treatment of rheumatoid arthritis. AB - Abatacept (CTLA4-Ig) is a novel fusion protein designed to modulate the T cell co stimulatory signal mediated through the CD28-CD80/86 pathway. Clinical trials have provided preliminary evidence of the efficacy of this compound in the treatment of rheumatoid arthritis. This review describes the molecular and biologic bases for the use of abatacept in rheumatoid arthritis and summarizes the current clinical data on its safety and effectiveness in this disease. PMID- 15833147 TI - Antiplatelet agents for prevention of pre-eclampsia and its consequences: a systematic review and individual patient data meta-analysis. AB - BACKGROUND: There is now good evidence that antiplatelet agents (principally low dose aspirin) prevent pre-eclampsia, a leading cause of morbidity and mortality for pregnant women and their babies. A Cochrane Review identified moderate, but clinically important, reductions in the relative risks of pre-eclampsia (19%), preterm birth (7%) and perinatal mortality (16%) in women allocated antiplatelets, rather than placebo or no antiplatelet.Uncertainty remains, however, about whether some women (in terms of risk) benefit more than others, what dose of aspirin is best and when in pregnancy treatment should ideally start. Rather than undertake new trials, the best way to answer these questions is to utilise existing individual patient data from women enrolled in each trial. METHODS/DESIGN: Systematic review with meta-analysis based on individual patient data. This involves the central collection, validation and re-analysis of thoroughly checked data from individual women in all the available randomised trials.The objective is to confirm that antiplatelet agents, given during pregnancy, will reduce the incidence of pre-eclampsia. The review will then determine the size of this effect, and whether antiplatelets delay the onset of pre-eclampsia or its impact on important outcomes for women and their babies. It will also explore whether the effect of antiplatelets differs by womens' risk profile; when commenced during pregnancy; and/or by dose. DISCUSSION: The PARIS (Perinatal Antiplatelet Review of International Studies) Collaboration has been formed to undertake the review. This will be the first individual patient data review in the perinatal field. Final results should be available by 2006-7. PMID- 15833155 TI - [How to cope with new challenges in the diagnosis and management of inborn errors of metabolism]. PMID- 15833156 TI - [Growth and growth hormone treatment--anything new?]. PMID- 15833146 TI - The role of the T cell in autoimmune inflammation. AB - T cells, in particular CD4+ T cells, have been implicated in mediating many aspects of autoimmune inflammation. However, current evidence suggests that the role played by CD4+ T cells in the development of rheumatoid inflammation exceeds that of activated proinflammatory T-helper (Th)1 effector cells that drive the chronic autoimmune response. Subsets of CD4+ T cells with regulatory capacity, such as CD25+ regulatory T (Treg) cells and Th2 cells, have been identified, and recent observations suggest that in rheumatoid arthritis the function of these regulatory T cells is severely impaired. Thus, in rheumatoid arthritis, defective regulatory mechanisms might allow the breakdown of peripheral tolerance, after which the detrimental Th1-driven immune response evolves and proceeds to chronic inflammation. Here, we review the functional abnormalities and the contribution of different T cell subsets to rheumatoid inflammation. PMID- 15833157 TI - [Mutation analysis of glycogen debrancher enzyme gene in five Chinese patients with glycogen storage disease type III]. AB - OBJECTIVE: Type III glycogen storage disease (GSD-III, McKusick 232400), is a rare autosomal recessive disorder, also known as Cori's or Forbe's disease. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (glycogen debrancher enzyme, GDE or amylogluco-sidase, AGL), which is responsible for the debranching of the glycogen molecule during catabolism. The AGL gene is located on chromosome 1p21 and contains 35 exons translated in a monomeric protein product. The clinical manifestations of GSD-III are represented by hepatomegaly, recurrent hypoglycemia, seizures, growth failure, dysmorphism, hyperlipidemia, raised transaminases and creatine kinase concentrations and, in a number of subjects, myopathy and cardiomyopathy. The hepatocellular adenoma, hepatocellular carcinoma, diabetes mellitus and liver fibrosis remain rare events. The diagnosis of debrancher deficiency was established by laboratory tests, electromyography (EMG), and muscle and liver biopsy. METHODS: We studied six GSD-III families after patients or parental consent and the clinical characteristics were documented. Analysis of 33 exons and part exon-intron boundaries of the AGL gene in patients and their parents were carried out by PCR and direct DNA sequencing. RESULTS: The clinical features included hepatomegaly, splenomegaly, recurrent hypoglycemia, hyperlipidemia, growth failure, raised transaminases and acidosis. Administration of epinephrine 2 hours after a carbohydrate meal could provoke normal rise of blood glucose in the affected individuals, but could not evoke any response after overnight fasting. Administration of raw-corn-starch could maintain normoglycemia and improve the disease condition. Mutation analysis for patient 1 was normal. Patient 2 had a compound heterozygote: a C-to-T transition at nucleotide 1294 (come from father, 1294C > T, L 298 L) in exon 8 and a G-to-T transition at nucleotide 4747 (from mother, 4747G > T, E1450X) in exon 34. Patient 3 had a compound heterozygote: a C-to-T transition at nucleotide 1294 (from father, 1294C > T, L 298 L) in exon 8 and a G-to-A transition at nucleotide -10 (from mother, -10G > A) in exon 3. Patient 4 was a homozygote: an insertion of a nucleotide CT into position +65 in exon 35 (4664 ins CT). Patient 5 had a compound heterozygote: a 8 bp deletion at nucleotide 2341 (from father, 2341delGCCATAGA, frameshift mutation) in exon 16 and a G-to-A transition at nucleotide 1559 (from mother, 1559G > A, R 387 Q) in exon 10. Patient 6 had a compound heterozygote: a T-to-G transition at nucleotide 1686 (from mother, 1686T > G, Y429 X) in exon 12 and a G-to-A transition at nucleotide 3742 (from father, 3742G > A, G 1115 R) in exon 26. CONCLUSION: GSD-III patients have variable phenotypic characteristics. Administration of raw-corn-starch can effectively improve the disease outcome. We identified 8 new mutations on AGL gene through nucleotide sequence analysis. PMID- 15833158 TI - [NEMO Delta 4-10 deletion of NEMO gene in Chinese incontinentia pigmenti cases]. AB - OBJECTIVE: Incontinentia pigmenti (IP) is a rare X-linked dominant disorder that affects ectodermal tissues. In IP, mutations in NEMO lead to the complete loss of NF-kB activation creating a susceptibility to cellular apoptosis in response to TNF-alpha. Recently, a second nonfunctional copy of the gene, Delta NEMO was identified, opposite in direction to NEMO. Almost 90% of IP whose gene mutation type had been recognized have a recurrent genomic deletion of exons 4-10 of the NEMO (IKK gamma) gene, called NEMO Delta 4-10, which is necessary to activate the NF-kB pathway. Therefore, PCR-based detection of the NEMO deletion is a diagnostic measurement for IP. This study sought to analyze the NEMO Delta 4-10 deletion in NEMO gene of Chinese IP cases. METHODS: Seven IP cases and part of their families totally 15 persons were enrolled in this study. The 7 IP cases were aged 41 days to 8 years. Among them 1 was male and 6 were female. Four cases had family history of IP, the other 3 were sporadic cases. Fifty healthy children without any congenital diseases were taken as normal control group. According to the gene characteristics of IP, by PCR measurement NEMO Delta 4-10 deletion in NEMO gene was tested with specific primers In2/JF3R, and NEMO Delta 4-10 deletion in pseudogene Delta NEMO was checked out by primers Rev-2/JF3R. RESULTS: Five out of the 7 tested cases (case 1, 2, 3, 4, and 6) showed NEMO Delta 4-10 deletion in NEMO gene. The mothers of case 1 and case 6, 1a and 6a, also suffered from this disease, and their results were just the same as their daughters. For pseudogene Delta NEMO only case 2 and case 4 were proved having NEMO Delta 4-10 deletion, while other cases and families had negative results. For normal control group, NEMO Delta 4-10 deletion was not found either in NEMO gene or in their pseudogene Delta NEMO. CONCLUSION: Incontinentia pigmenti in most cases were caused by NEMO Delta 4-10 deletion in NEMO gene. PMID- 15833159 TI - [Oral glucose tolerance and insulin release test in 52 cases of obese and overweight children]. AB - OBJECTIVE: To assess the state of glucose metabolism and beta-cell function in obese and overweight children. METHODS: Levels of glucose and insulin were detected during oral glucose tolerance test in 52 obese and overweight children aged 11.3 +/- 1.8 years with body mass index (BMI) 30.2 +/- 19.2 kg/m(2). Insulin resistance index (IR = FIN x FPG/22.5), insulin sensitivity index (IS = 1/FIN x FPG) and ratio of insulin increment to glucose increment at 30' (I(30)-I(0)/G(30) G(0)) post oral glucose were measured. (FIN = fasting insulin. FPG = fasting plasma glucose). The IR, IS and the ratio post oral glucose were compared among groups with varying BMI and between groups of impaired glucose tolerance (IGT) and control. Serum triglyceride determination and B ultrasonography of liver were performed. RESULTS: (1) one patient with type 2 diabetes (1.9%) and 5 patients with IGT (9.6%) were found. (2) IR (> or = 2.8) was observed in 76.9% of the cases. (3) The IR, IS and their ratio showed no difference between the compared groups. (4) IR and IS did not show significant difference but there was significant difference in ratio between the IGT and control group. (5) Increased serum triglyceride and fatty liver were noted in 36.5% and 53.3% of the cases, respectively. CONCLUSION: The results indicated that insulin resistance and reduced insulin sensitivity in obese and overweight children are common, and these changes seemed not to correlated with the varying degree of BMI. Beta-cells function was obviously impaired in obese children with IGT and disorder of lipid metabolism exists in many obese and overweight children revealed. PMID- 15833160 TI - [Thyroid autoantibodies in children with Turner's syndrome]. AB - OBJECTIVE: Turner's syndrome (TS) is characterized by the absence of an X chromosome or the presence of a structurally abnormal X chromosome in a phenotypic female. It was recently reported that autoimmune thyroiditis (AIT) was found in 38% of white patients with TS, and few studies in this aspect have been conducted in China. The purpose of this study was to determine the frequency of AIT among TS patients and risk factors for development of thyroid dysfunction in Chinese children with TS. METHODS: Serum antithyroglobulin antibody (TgAb), thyroperoxidase antibody (TPOAb) and thyroid function (T(3), T(4) and TSH) of 24 children with TS (mean age 12.9 +/- 2.4 years, range 4.8 - 16.8 years) were assessed. Their karyotype distribution was as follows: thirteen patients with 45, XO kayrotype, eight patients with structurally abnormal X chromosome, two with X mosaic kayrotype and one with 46, XX. Techniques including radioimmunoassy and elctro-chemiluminescence immunoassy were used in this study. All TS children were divided into two groups. Group one was thyroid autoantibodies (TAA)-positive group, the levels of TgAb and/or TPOAb in them were higher than the normal levels (TgAb < 30%, TPOAb < 20%), respectively, and the remaining patients were assigned into TAA-negative group. RESULTS: Seven of the 24 (29%) patients had higher levels of TgAb and TPOAb than the normal values (< 30% and < 20%). The level of serum TSH [6.1 (3.6-100.0) mU/L] in TAA-positive group was significantly higher than that [3.9 (1.7-7.9) mU/L] in TAA-negative group (P < 0.05). The frequency of hypothyroidism or subclinical hypothyroidism in TAA-positive group (5/7) was higher than that in TAA-negative group (3/17) (P < 0.05). CONCLUSION: The positive rate of serum TAA in children with TS was 29%. About 70% TS children with positive serum TAA developed hypothyroidism or subclinical hypothyroidism. The results have provided the basis for regular follow-up assessment of thyroid autoantibodies and thyroid function in children with TS, and these measures are of importance for timely diagnosis of thyroid dysfunction and application of appropriate treatment. PMID- 15833161 TI - [Functional examination of growth hormone-insulin-like growth factor axis in short stature children]. PMID- 15833162 TI - [Summary of the conference of endocrinologic, hereditary and metabolic diseases of the Society of Pediatrics, Chinese Medical Association]. PMID- 15833163 TI - [Summary of 2004 national symposium on hematuria and chronic renal failure in children]. PMID- 15833164 TI - [Summary of the first national symposium for chiefs of pediatric departments organized by the Pediatric Society, Chinese Medical Association]. PMID- 15833165 TI - [Summary of national conference of pediatric respiratory diseases]. PMID- 15833166 TI - [Effect of glucocorticoid on glucocorticoid-resistant children with primary nephrotic syndrome]. AB - OBJECTIVE: Glucocorticoid (GC) is the first therapeutic choice of primary nephrotic syndrome (PNS). The response to GC treatment is an important indicator for the outcome of PNS children. Children with GC-resistant PNS present with incomplete or no response to GC, and may herald the progression to end-stage renal failure. However, the detailed mechanism of GC-resistance or GC-sensitive effect in these PNS children has not been clearly elucidated. The previous study by the authors indicated that there was increased expression of GR beta in PBMCs in GC-resistant children with PNS, and the over expression of GR beta resulted in GC resistance via influencing the ability of GR alpha nuclear translocation. To elucidate the relationship between GR beta expression in renal and in PBMCs and the effect of glucocorticoid on glucocorticoid-resistance children with PNS, the expression of GR alpha and GR beta in renal tissue and in PBMCs were detected by immunohistochemistry. METHODS: Forty children with PNS were divided into two groups, GC-resistant group(20) and GC-sensitive group(20), the expression of GR alpha and GR beta in renal intrinsic cells and in PBMCs were measured with the immunohistochemistry technique. A semiquantitative score was used to evaluate the injury degree of the glomeruli and tubulointerstitium. RESULTS: Compared with GC sensitive group, the glomerular pathologic scores (6.91 +/- 1.98) and renal tubular pathologic scores (7.12 +/- 1.62) in GC- resistant group were significantly different (P < 0.01, respectively). GR alpha expressions of renal tissue and PBMCs were higher in the control group (58.3 +/- 2.6, 59.1 +/- 7.2) than those in the GC-sensitive group (40.2 +/- 7.2 and 36.6 +/- 5.1, P < 0.01, respectively) and GC-resistant group (35.0 +/- 8.2 and 36.4 +/- 6.6, P < 0.01, respectively). GR beta expressions of renal tissue and PBMCs were higher in the GC-resistant group (13.8 +/- 3.0 and 12.1 +/- 4.1) and in the GC-sensitive group (6.5 +/- 1.9 and 5.9 +/- 1.0) than that in control group (2.3 +/- 0.4 and 3.2 +/- 1.1, P < 0.01, respectively). GR beta expressions in renal tissue and PBMCs were higher in the GC-resistant group than that in the GC-sensitive group (P < 0.01). Compared with control group, GR beta expressions in PBMCs and in renal tissue were lower than those in mild renal lesion group (5.4 +/- 2.8, 6.46 +/- 2.50), midmedium renal lesion group (8.7 +/- 2.4 and 11.4 +/- 3.7) and (17.1 +/- 0.4 and 18.7 +/- 0.7) in severe renal lesion group (F = 5.8, 15.6, P < 0.01, respectively). GR beta expression of PBMCs had a positive correlation with GR beta expression of renal intrinsic cells (r = 0.651, P < 0.01). GR beta expressions by PBMCs and renal intrinsic cells were positively correlated with renal pathologic scores (r = 0.579 and 0.623, P < 0.01, respectively). CONCLUSION: GC-resistant children with PNS were related to the increased GR beta expression in PBMCs and renal intrinsic cells. There was no correlation between the GR alpha expressions in PBMCs and in renal intrinsic cells. Increased GR beta expression might decrease the effect of GC via inhibiting the activity of GR alpha. PMID- 15833167 TI - [Diagnosis and treatment of chronic pulmonary aspergillosis in 4 children]. AB - OBJECTIVE: Chronic pulmonary aspergillosis is different from acute pulmonary aspergillosis in clinical picture, radiogram, diagnostic procedures and prognosis. Four patients with chronic pulmonary aspergillosis had been misdiagnosed as having pneumonia or pulmonary tuberculosis for a long time before admission to the hospital. The purpose of this report was to summarize the clinical manifestations and laboratory findings for correct diagnosis of chronic pulmonary aspergillosis. METHODS: Four patients with chronic pulmonary aspergillosis seen between October 2002 and October 2004 were retrospectively studied. Their clinical manifestations, chest radiographic feature, immune status, diagnostic procedure, therapy and prognosis were reviewed. RESULTS: The chief complaints of these patients were chronic cough and fever for 3 to 12 months. Chest wall abscess developed in the late course in case 1 and 4. Fine moist rales were heard and hepatosplenomegaly was found in case 1 and 2. No abnormal sign was found in case 3 and 4. Chest radiographic feature: lobar consolidation with adjacent pleural thickening was present in all cases. In early phase, solitary or multiple small nodules were found in 2 cases. Case 1-3 had normal IgG, IgM, IgA, IgE, T Cell subsets and NBT test. Case 4 had chronic granulomatous disease. Etiologic evidences: culture was positive for Aspergillus (A.) fulmigatus in sputum and in chest wall abscess in case 1 and 4; for A. niger in sputum and spore existing in lung tissue in case 2; for A. fulmigatus in sputum and hypha existing in lung tissue in case 3. All patients were treated with combination of amphotericin B and itraconazole. Their symptoms were controlled 10-30 d after treatment. In case 1 the disease relapsed 6 months later and the patient died at last due to giving up treatment by his parents. Case 2 was free of symptom for 12 months and his chest radiographic lesion disappeared completely 6 months later. Treatment of case 4 was given up. Case 3 continued to receive treatment and observation. CONCLUSION: Chronic pulmonary aspergillosis should be considered in children with long period fever and cough and lobar consolidation associated with adjacent pleural thickening or with nodular infiltration. The diagnosis of chronic pulmonary aspergillosis depended on identification of aspergillus from sputum or lung tissue. Combined amphotericin B and itraconazole might control the disease. PMID- 15833168 TI - [Protective effects of amygdalin on hyperoxia-exposed type II alveolar epithelial cells isolated from premature rat lungs in vitro]. AB - OBJECTIVE: To analyze the effect of hyperoxia on the proliferation and surfactant associated protein messenger RNA levels of type II alveolar epithelial cells (AECIIs) of premature rat, and to investigate the effect of amygdalin on the change resulted from hyperoxia in AECIIs isolated from premature rat lung in vitro. METHODS: The lung tissue of 20-day fetal rat was digested by trypsin and collagenase. AECIIs and lung fibroblasts (LFs) were isolated and purified at different centrifugal force and different adherence, then cultured. The nature of the cultures was identified by cytokeratin staining, vimentin staining and transmission electron micrography. For establishing hyperoxia-exposed cell model, purified AECIIs were cultured for 24 hours after culture flasks were filled with 95% oxygen-5% CO2 at 3 L/min for 10 min, and then sealed. Oxygen concentrations were tested in CYS-1 digital oxygen monitor after 24 hours of exposure. A sample was discarded if its oxygen concentration was < 90%. Cell proliferating vitality was examined by MTT assay after treatment with amygdalin at various concentrations. DNA content, protein expression of proliferating cell nuclear antigen (PCNA) and mRNA levels of SPs of AECIIs were analyzed with flow cytometric assay, Western blot and reverse transcription polymerase chain reaction (RT-PCR) respectively after 24 hours of air or hyperoxia exposure in the presence or absence of 200 micromol/L amygdalin. RESULTS: Excellent yields of highly purified, culturable AECIIs could be obtained from 20-day fetal lungs. The expression of cytokeratin in AECIIs was positive and that of vimentin negative by immunocytochemistry. Those, however, in LFs were just opposite. Lamellar bodies in purified AECIIs were revealed by transmission electron micrography. The established hyperoxia-exposed cell model assured the oxygen concentrations of culture flasks more than 90%. Amygdalin at the concentration range from 50 micromol/L to 200 micromol/L stimulated the proliferation of AECIIs in a dose dependent manner; however, at the concentration of 400 micromol/L inhibited the proliferation of AECII. Flow cytometric analysis showed that the apoptosis rate and G0/G1 phase percentage increased significantly (P < 0.01), S phase and G2/M phase percentage decreased significantly (P < 0.01), in hyperoxia group compared with that of air group. The apoptosis rate of air plus 200 micromol/L amygdalin group, compared with air group, was not significantly different (P > 0.05); however, G0/G1 phase percentage decreased markedly, S phase percentage increased significantly, G2/M phase percentage did not significantly change (P > 0.05). The apoptosis rate of hyperoxia plus 200 micromol/L amygdalin group was not significantly different (P > 0.05) from that of hyperoxia group, S phase and G2/M phase percentage increased significantly (P < 0.01), G0/G1 phase percentage decreased significantly (P < 0.01). Western blot analysis showed that the protein expression levels of PCNA in all group was significantly different, in turn, hyperoxia group < hyperoxia plus 200 micromol/L amygdalin < air group < air puls 200 micromol/L amygdalin (P < 0.01). SPs mRNA levels were significantly decreased in hyperoxia group, as compared with air group (P < 0.01). After amygdalin was added, SPs mRNA levels were elevated in air plus amygdalin group and hyperoxia plus amygdalin group, as compared with hyperoxia group (P < 0.01, P < 0.05, respectively), but compared with air group, SP mRNA levels were not significantly elevated (P > 0.05). CONCLUSION: AECIIs of premature rats were isolated, purified and cultured successfully. Hyperoxia-exposed cell model was established in AECIIs of premature rat in this experiment. Amygdalin promotes the proliferation of premature rat AECII exposed to air or hyperoxia, the concentration of amygdalin with the best effect was 200 micromol/L. Hyperoxia inhibited the proliferation and decreased SPs mRNAs levels in AECIIs in vitro, which may contribute to hyperoxia-induced lung injury in premature rats. Amygdalin could inhibit the changes of SPs mRNAs levels and cell proliferation of AECIIs resulted from hyperoxia and may play partial protective role in hyperoxia-induced premature lung injury. PMID- 15833169 TI - [Effects of Bacillus Calmette-Guerin vaccination on immune functional development of splenic T cell in neonatal mice]. AB - OBJECTIVE: Almost every neonate receives Bacillus Calmette-Guerin (BCG) vaccination in China. The authors' previous study showed that BCG promoted cord blood monocyte-derived dendritic cells maturation and induced high level of interleukin (IL)-10, medium level of interferon (IFN)-gamma, but low level of IL 4 production by cord naive T cells. The experiments in the present study were designed to explore the effects of neonatal BCG vaccination on immune functional development of splenic T cells in mice in vivo. METHODS: Neonatal BALB/c mice were inoculated with BCG intraperiotoneally. Four weeks later, spleen cells of mice were isolated and surface molecular markers of CD4, CD25 and CD44 and intracellular IFN-gamma, IL-10, and IL-4 in CD3(+) T cells were detected by flow cytometry. Furthermore, mRNA expression of transcription factor T-bet, Foxp3 and GATA-3 were analyzed by RT-PCR. RESULTS: The percentage of total CD4(+) T cells decreased [(23.50 +/- 2.59)% vs. (47.38 +/- 10.41)%, P < 0.01] but the percentage of CD25(+) [(24.92 +/- 2.74)% vs. (20.27 +/- 2.85)%, P < 0.05] and CD44(+) [(89.29 +/- 2.56)% vs. (82.98 +/- 5.51)%, P < 0.05] T cells in CD4(+) T cells was higher in BCG-vaccinated mice than that in controls. Meanwhile, the percentage of IFN-gamma positive [(6.52 +/- 2.40)% vs. (3.13 +/- 2.03)%, P < 0.05] and IL-10 positive [(14.81 +/- 3.65)% vs. (10.90 +/- 1.61)%, P < 0.05] but not IL-4 positive [(1.17 +/- 0.46)% vs (1.51 +/- 0.75)%, P > 0.05] cells in CD3(+) T cells of BCG-vaccinated mice was significantly higher than that of non-BCG-vaccinated mice. In comparison with BCG-naive mice, T-bet was significantly high in BCG vaccinated mice [T-bet/beta-actin 0.44 +/- 0.11 vs. 0.28 +/- 0.06, P < 0.05], but there was no significant difference in GATA-3 [GATA-3/beta-actin 0.46 +/- 0.08 vs. 0.50 +/- 0.10,P > 0.05] and Foxp3 [Foxp3/beta-actin vs. 0.27 +/- 0.11 and 0.30 +/- 0.16, P > 0.05] mRNA expression between the two groups. CONCLUSION: Neonatal BCG vaccination could induce strong Th1 but weak Th2 response as reported previously. Though neonatal BCG vaccination was not capable of inducing CD4(+)CD25(+) regulatory T cell response with Foxp3 expression, it caused increase of IL-10(+) CD3(+) cells which might represent some regulatory T cells producing IL-10. PMID- 15833170 TI - [Examination of infant bone status with quantitative ultrasound at birth]. AB - OBJECTIVE: Quantitative ultrasound (QUS) is a new method of evaluating Children's bone status, including bone mineral density and bone strength. The bone nutrition during fetal and early neonatal period is very important for the human bone development of whole life. The objective of this study was to evaluate the clinical application of QUS for newborn infants and to obtain the QUS data for normal neonates including premature infants. METHODS: An ultrasound bone sonometer, Omnisense, produced by Sunlight company of Israel, was applied to measure the bone speed of sound (SOS) of tibia in 157 neonates including 68 premature infants in the first week of life. RESULTS: (1) No significant difference in SOS was found between male (n = 88, SOS = 2968 +/- 115 m/s) and female infants (n = 69, SOS = 2956 +/- 105 m/s) (P = 0.524). The SOS of premature infants (n = 68, mean gestational age 33.0 +/- 2.5 weeks) and full-term infants (n = 89, mean gestational age of 39.4 +/- 1.3 weeks) were 2935 +/- 96 m/s and 2984 +/- 116 m/s, respectively, at birth and there was significant difference between them (P = 0.005). (2) There were significant differences of SOS in neonates who were born in different seasons (F = 4.377, P = 0.005); the significant difference remained (F = 3.933, P = 0.010) after the influences of gestational age and birth weight were eliminated. The SOS in neonates born in spring (March, April and May) and summer (June, July and August) were significantly lower than that of those born in autumn (September, October and November) and winter (December, January and February). The SOS in neonates born in summer was about 2.3% (75/2999) was lower than that of those born in winter. (3) Significant difference of SOS was observed between neonates with different birth weight [< 1500 g (n = 11), SOS = 2968 +/- 115 m/s; and > 2500 g (n = 86), SOS = 2980 +/- 113 m/s; P = 0.042]. (4) Significant correlations were found between SOS and gestational age (r = 0.270, P = 0.005), and between bone SOS and birth weight (r = 0.232, P = 0.015) in appropriate for gestational age (AGA) infants (n = 109); however, no such significant correlations were found in small for gestational age (SGA) infants or large for gestational age (LGA) infants. Multiple regression analysis showed that gestational age and the birth season were two important factors which may contribute to bone SOS of neonates at birth (n = 157, F = 8.515, P < 0.001, adjusted R(2) = 0.141), when the analysis was carried out with SOS as dependent variable and gestational age, birth weight, chronological age, calf length and the birth season as independent variables. CONCLUSION: QUS is a new method which is suitable for evaluating the bone status of neonates and it is free of radiation, non-invasive, the machine is portable and easy to manipulate at infant bed side. The present study suggests the need for particular care for the bone status in premature infants and supplement of vitamin D in pregnant women. PMID- 15833171 TI - [T-type calcium channel gene-CACNA1H is a susceptibility gene to childhood absence epilepsy]. AB - OBJECTIVE: Childhood absence epilepsy (CAE) is one of the most frequently recognized syndromes among the idiopathic generalized epilepsies (IGEs). It is considered to be a hereditary disease. The possible inheritance pattern of CAE is polygenic. The genes responsible for CAE, however, have not yet been identified. The aim of this study was to further investigate based on the authors' recent work whether or not T-type calcium channel gene-CACNA1H is a susceptibility gene to childhood absence epilepsy. METHODS: The authors conducted the mutation screening of the exons 6-12 and the nearby partial introns of the CACNA1H gene using the method of direct sequencing of PCR products in 48 newly found CAE patients. RESULTS: The authors found 13 single nucleotide polymorphisms (SNPs). They also found 4 mutations which only existed in CAE patients. Both G773D and H515Y mutations were heterozygous. The mutation of H515Y has never been reported previously. The patient inherited the mutation from his mother. The authors found two CAE patients with the mutation of G773D previously. This is the third time that the authors found one more CAE family with this G773D mutation, and the patient with the mutation G773D inherited the mutation from his father. CONCLUSION: T-type calcium channel gene-CACNA1H might be a susceptibility gene to childhood absence epilepsy. PMID- 15833172 TI - [Possible association between HLA-HRB1 and DQB1 genes frequency and susceptibility or resistance to Helicobacter pylori infection in Kunming Yi ethnic group children]. AB - OBJECTIVE: To study if there is any association between frequency of HLA-DRB1 and DQB1 genes and susceptibility or resistance to Helicobacter pylori (Hp) infection among children of Yi ethnic group in Kunming for understanding the immunogenetic features of the digestive diseases associated with Hp infection. METHODS: Peripherial blood samples were collected from 156 children of Yi ethnic group in a primary school in Kunming city by cluster sampling and the blood Hp-IgG tests (ELISA) were performed. The samples were divided into two groups (Hp-IgG-positive group and Hp-IgG-negative group) according to the blood Hp-IgG test results. There were 61 children in Hp-IgG-positive group and 95 children in Hp-IgG negative group. Forty children who were chosen from each group by simple random sampling underwent (13)carbon-urea breath test ((13)C-UBT). Thirty-three children who were Hp-IgG-positive and (13)C-UBT-positive were defined as currently Hp- infected group; 39 children who were Hp-IgG-negative and (13)C-UBT-negative were defined as Hp-non-infected group. DNA specimens were extracted from the lymphocytes of their peripheral blood samples. HLA-DRB1 and DQB1 DNA typing was performed by using polymerase chain reaction with sequence specific primers (PCR SSP). HLA-DRB1, DQB1 allelic frequency distribution among currently Hp infected and non-infected children was compared. RESULTS: HLA-DRB1 * 12 gene frequency among children in Hp non-infected group was higher than that in the currently Hp infected group (42.31% vs. 14.52%, P < 0.001, Pc < 0.012); however, HLA-DRB1 * 11 gene frequency in the Hp-non-infected group was lower than that in the currently Hp-infected group (3.85% vs. 12.9%, P < 0.05, Pc > 0.05). HLA-DQB1 * 0301 gene frequency in the Hp non-infected group was higher than that in the currently Hp infected group (55.13% vs. 32.26%, P < 0.007, Pc < 0.05); however, HLA-DQB1 * 04 gene frequency in the Hp non-infected group was lower than that in currently Hp infected group (2.56% vs. 11.29%, P < 0.05, Pc > 0.05). CONCLUSIONS: HLA-DRB1 * 12 and HLA-DQB1 * 0301 gene may be associated with protection against Hp infection in Kunming Yi ethnic group children. Further studies with larger sample size are needed to clarify if HLA-DRB1 * 11 and HLA-DQB1 * 04 are associated with susceptible gene to Hp infection. PMID- 15833173 TI - [Preliminary study on cytomegalovirus infection in children with primary glomerular disease]. PMID- 15833174 TI - [Nutritional intake and weight gain of very low birth weight infants during hospitalization]. PMID- 15833175 TI - [Endoscopic findings of gastrointestinal involvement in patients with Henoch Schonlein purpura with abdominal pain as the first symptom]. PMID- 15833176 TI - [Clinical effects of montelukast sodium in treatment of allergic rhinetis in children]. PMID- 15833177 TI - [A case of birch pollen-related food allergy syndrome]. PMID- 15833178 TI - [Hallervorden-Spatz disease of childhood onset type, a report of 2 cases]. PMID- 15833179 TI - [Treatment of congenital chylous ascites using total parenteral nutrition and somatostatin: a case report]. PMID- 15833180 TI - [Steroid-resistant nephrotic syndrome and NPHS2 gene]. PMID- 15833181 TI - [Rippling muscle disease in children]. PMID- 15833182 TI - [Several questions concerning the article "Latex allergen reactivity and antigenic cross reactivity in children with allergic diseases"]. PMID- 15833183 TI - [Diagnostic value of plasma concentration of pro-brain natriuretic peptide in congestive heart failure in pediatric patients with ventricular septal defects]. AB - OBJECTIVE: The value of plasma brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) can reflect cardiac function and therefore can be used for diagnosing congestive heart failure (CHF) and evaluating cardiac function. There are few reports, however, on the value of BNP and NT-proBNP in pediatric cases of congenital heart defect. The aim of this study was to assess the value of plasma NT-proBNP in the diagnosis of CHF and evaluation of cardiac function in pediatric patients with ventricular septal defect (VSD). METHODS: Fifty-one patients with VSD aged from 2 months to 2 years old (mean 7.9 months) were enrolled. According to the modified Ross Score, the patients were divided into three groups, no CHF group (20 patients), mild CHF group (18 patients) and moderate to severe CHF group (13 patients). Fifteen age-matched normal children were used as controls. Plasma NT-proBNP was measured using enzyme immunoassay. All patients had complete echocardiographic study, including measurement of left ventricular end diastolic volume index (LVEDVI), left ventricular end systolic wall stress (LVSEWS), heart rate corrected mean velocity of circumferential fiber shortening (mVcFc), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), and contractility index (Con). The correlation between plasma NT-proBNP level and modified Ross Score and echocardiographic cardiac functional indexes was determined. The sensitivity, specificity and ROC curve of plasma NT-proBNP for diagnosing CHF was studied. RESULTS: Plasma NT-proBNP was positively correlated with modified Ross Score (r = 0.75, P < 0.01). Plasma NT-proBNP concentration in moderate to severe CHF group (2061 +/- 908) fmol/ml was significantly higher than that of mild CHF group (810 +/- 335) fmol/ml, and Plasma NT-proBNP concentration in mild CHF group was higher than that in no CHF group (309 +/- 68) fmol/ml. 97.14% of normal controls and subjects in no CHF group had their plasma NT-proBNP below 400 fmol/ml. 83.3% of children in mild CHF group had their plasma NT-proBNP between (400-1400) fmol/ml while in moderate and severe CHF group 84.6% of children had their plasma NT-proBNP beyond 1400 fmol/ml. Plasma NT-proBNP was also positively correlated with LVEDVI and LVSEWS. There was no correlation among mVcFc, LVEF, LVFS, Con and plasma NT-proBNP concentration. Using plasma NT-proBNP concentration > or = 400 fmol/ml as cut point for diagnosing CHF, the sensitivity was 89.3%, the specificity was 91.2%, and the area under the ROC curve was 0.944. CONCLUSIONS: Plasma NT-proBNP level could be used to assess cardiac function and diagnose CHF in pediatric patients with VSD. PMID- 15833184 TI - [Clinical analysis and follow-up study of postural orthostatic tachycardia syndrome in 28 pediatric cases]. AB - OBJECTIVE: Lightheadedness, dizziness, chest discomfort, headache, pallor, palpitation, blurred vision, fatigue, and syncope are main symptoms of postural orthostatic tachycardia syndrome (POTS), but they are also common problems in clinical pediatrics. Besides, most of POTS cases have normal findings in routine examinations on cardiac and central nervous system. Therefore, they are often unable to be correctly diagnosed and treated in time. Since head-up tilt test (HUT) was introduced to diagnose unexplained syncope in children, it was discovered that the most common pattern of unexplained syncope in children was vasovagal syncope. But in the course of the investigation, we and other groups identified a large subgroup of patients who had a less severe form of orthostatic intolerance characterized by postural tachycardia, lightheadedness, dizziness, and chest discomfort, etc. This disorder has become generally known as the POTS, which in children is not rare, but its report was not seen in China so far. The aim of this article is to recognise the clinical pictures of POTS in children and to explore its diagnostic criteria and therapeutic protocol. METHODS: All 28 pediatric patients of POTS who met the diagnostic criteria were selected to be the observation group. Age and sex distribution were observed, and the duration of symptoms, baseline heart rate and blood pressure were also recorded. The frequency of symptoms, the rate of misdiagnosis and the efficiency of therapeutic protocol were also analysed. RESULTS: Twenty-eight patients were diagnosed as POTS, occupying 31.8% of all unexplained syncope or lightheadedness in children. Eleven were males and 17 females. The ratio of male to female was 1:1.5. All the 28 cases were from 6 to 16 years old, 5 of which were < 10 years and 23 were from 10 to 16 years old, including 10. The duration of symptoms of POTS in children was from 1 month to 6 years. The average was about 13 months, and more than one half were in 6 months. The most common orthostatic symptoms were lightheadedness or dizziness, syncope, chest discomfort, pallor and blurred vision associated with nausea and/or vomiting. During HUT or standing, an increase in heart rate > 35 beats per minute within 10 minutes was the most common finding, and some were associated with > 120 beats per minute within 10 minutes after HUT or standing up. The average time of the occurrence of abnormal responses was about 5 minutes. Especially, we found only 36% of patients were diagnosed with standing up test. Most of them were diagnosed with HUT. POTS in children was misdiagnosed as epilepsy or myocarditis in 43% patients. Multiple treatment protocol including health education and supportive, physical and medical therapy were most helpful in children with POTS. CONCLUSION: POTS is commonly seen in school-aged girls. The common symptoms include dizziness, syncope and chest distress. HUT is an important tool in its diagnosis. A comprehensive therapeutic regimen is recommended in the treatment. PMID- 15833185 TI - [A tal-1 deletion as real-time quantitative polymerase chain reaction target for detection of minimal residual disease in T-lineage acute lymphoblastic leukemia]. AB - OBJECTIVE: Hematologic relapse remains the greatest obstacle to the cure of acute lymphoblastic leukemia (ALL), especially T-lineage acute lymphoblastic leukemia (T-ALL) in children. Recent studies have shown that patients with increased risk of relapse can be identified by measuring residual leukemic cells, called minimal residual disease (MRD), during clinical remission. Current polymerase chain reaction (PCR) methods, however, for measuring MRD are cumbersome and time consuming. To improve and simplify MRD assessment, the author developed a real time quantitative PCR (RQ-PCR) assay for the detection of leukemic cells that harbor the tal-1 deletion. In addition, the author discussed the significance of MRD levels at different stages in treatment and prognosis of children with T-ALL. METHODS: A total of 50 consecutively enrolled patients with T-ALL were analysed for detection of leukemic cells harboring the most common tal-1 deletion. Serial dilutions of leukemic DNA were studied to find the sensitivity of detection with RQ-PCR assay. The MRD of 28 samples in clinical remission from 10 patients were quantified by RQ-PCR assay and limiting dilution assay. The results detected by both methods were compared statistically with correlation analysis. RESULTS: (1) A total of 10 patients presented tal-1 deletion involving the sildb1 breakpoint rearranged to tal1db1 in 50 cases with T-ALL. The breakpoints of relapsed samples are the same as those of the corresponding diagnostic samples; (2) The RQ-PCR assay had a sensitivity of detection of one leukemic cell among 100,000 normal cells. In 24 samples, MRD levels > 10(-5) could be detected with both methods. The percentages of leukemic cells measured by the two methods correlated well (r = 0.898, P < 0.001); (3) The MRD levels of 3 patients out of the 8 cases undergoing disciplinary regimen were over 10(-4) at the end of induction chemotherapy. They all relapsed in bone marrow during chemotherapy. The higher the MRD levels, the earlier the relapse. The other 5 patients with MRD levels < 10(-4) had been relapse-free survival (RFS) for 4-59 months, one of whom with increased MRD levels > 10(-4) for twice at the continuation stage had been RFS for 27 months till now. CONCLUSIONS: The sildb1-taldb1 deletion presents in 20% of T-ALL, and is an ideal PCR marker for its specificity, uniform and stability; The tal-1 RQ-PCR can be used for the rapidly, sensitively and accurately quantitative assessment of MRD in T-ALL with the tal-1 deletion. MRD levels at different stages of chemotherapy have different significance in prognosis and treatment. PMID- 15833186 TI - [Prophylaxis and treatment of chronic graft versus host disease]. AB - OBJECTIVE: Chronic graft versus host disease (cGVHD) is the most common late complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and it represents the major cause of mortality in long-term survivors. Over the past decade, although conventional therapy has achieved complete responses in approximately 50% of patients, the prophylaxis and treatment of cGVHD are still not satisfactory. In the late years, utilization of new immunosuppressant such as tacrolimus (FK506), mycophenolate mofetil (MMF) on cGVHD improved the curative effects. This study tried to analyze the results of combination of methylprednisolone (MP), MMF and FK506 or cyclosporine A (CSA) as immunosuppressive therapies for cGVHD and to explore the effective regimen for children. METHODS: Forty-five patients received allo-HSCT. Among them 32 received UCBT and 13 received PBSCT. The conditional regimen mainly consisted of busalphan, cyclophosphamide, antihuman thymocyte globulin, fludarabin, melphalan, thiotepa and total lymph node irradiation. Prophylaxis of GVHD consisted of CSA, MP and MMF. Patients with cGVHD received a regimen with combination of MP, MMF and FK506 or CSA. RESULTS: Seventeen out of 32 patients who received UCBT were engrafted. while 9 out of 13 patients who received PBSCT were engrafted. Nine cases of the 30 engrafted patients developed cGVHD (morbidity 30%). Among the 17 patients who received UCBT, 3 developed cGVHD (18%). Among the 13 patients who received PBSCT, 6 developed cGVHD (46%). Six cGVHD continued from aGVHD (6/9). One patient was given CSA plus MMF, and 8 were given three-drug regimen with MP, MMF and FK506. The overall response rate was 100%. Two patients died of CMV-IP or septicemia (mortality 20%). Seven (78%) patients survived (event free survival, EFS) longer than 3 years. The side effects included hepatotoxicity, nephrotoxicity, hypertension, articular capsulitis and arrhythmia. The main complication and the major causes of death were infection. CONCLUSION: The incidence of cGVHD is low in children. The incidence of cGVHD after PBSCT is higher than that after UCBT. aGVHD is a highly dangerous factor. Combined therapy of MP plus MMF and FK506 or CSA is safe and effective for the treatment of cGVHD in children. PMID- 15833187 TI - [Umbilical cord blood transplantation for patients with beta-thalassemia major]. AB - OBJECTIVE: The beta-thalassemia major is a common hereditary hematology disease in southern China. The combination of blood transfusion and iron chelation is now the reference treatment. The allogeneic hematopoietic stem cell transplantation is the only curative therapy for beta-thalassemia major. In this study the investigators observed and evaluated the effects of umbilical cord blood transplantation (UCBT) for patients with beta-thalassemia major. METHODS: Twelve cases of beta-thalassemia major aged from 1.3 to 8.3 years (8 male and 4 female) received UCBT. Eleven of the twelve donors were siblings and one was unrelative. Eight patients received no antigen and four patients received two antigen disparate grafts. According to the Pesaro's classification for thalassemia, 10 patients were at grade I or II, and 2 were at grade III. The HLA-identical patients accepted the conditioning regimen consisting of busulfan, cyclophosphamide and antithymocyteglobulin. The HLA-mismatched patients accepted the conditioning regimen consisting of hypertransfusions, continuous iv desferrioxamine, hydroxyurea, fludarabine, busulfan, cyclophosphamide and antithymocyteglobulin. The harvest stem cells contained 3.63 - 16.0 x 10(7)/kg of nucleated cells, 0.11 - 1.03 x 10(6)/kg of CD(34)(+) cells and 0.17 - 1.18 x 10(5)/kg of colony-forming-unit-granulocyte macrophages. Cyclosporine alone or in combination with mycophenolate mofetil (MMF) was given for acute graft-versus host disease (aGVHD) prophylaxis. RESULTS: Of the 12 patients, 10 were engrafted. Ten patients had neutrophil recovery (> 0.5 x 10(9)/L) and seven patients had platelet recovery (> 50 x 10(9)/L). The median time was 18.1 and 57.3 days, respectively. Seven patients had disease-free survival (DFS) at a median follow up of 23 months (range 4 - 63 months). Three patients had rejection and autologous hematopoitic reconstitution. Two patients were not engrafted. One patient acquired severe aplastic anemia, another patient died of severe infection. The incidences of grade I and grade II aGVHD were 60% (6/10) and 40% (4/10), respectively. There were no long-term complications in the disease free survivors. CONCLUSIONS: Grade I-II beta-thalassemia major patients receiving sibling UCBT had high DFS. UCBT is an effective way to treat beta-thalassemia major. PMID- 15833188 TI - [Comparative study on bone marrow megakaryocytes in children with thrombocytopenic purpura, aplastic anemia and myelodysplastic syndrome]. AB - OBJECTIVE: Thrombocytopenic hemorrhage is one of the major appearance in pediatric hemorrhagic diseases, in which, idiopathic thrombocytopenic purpura (ITP) is the most common disease. Thrombocytopenia is the earliest phenomenon or the only one in certain phases of hemorrhagic diseases, such as ITP, aplastic anemia (AA) and myelodysplastic syndrome (MDS). By now, the pathogenesis of thrombocytopenia in different diseases has not been clearly determined. At present, it is very difficult to diagnose these diseases and estimate their prognosis with current clinical data. In this study, morphological characteristics and hematopoiesis function of bone marrow megakaryocyte in pediatric patients with ITP, AA and MDS were observed and the cause and mechanism of different thrombocytopenias were analyzed. METHODS: There were 16 children with ITP, 17 with AA and 16 with MDS in this study. CD41 McAb immunohistochemical technique was used to detect micromegakaryocyte on bone marrow smears. Plasma clot culture and CD41 McAb immunohistochemical technique were used for the MK colony forming assay. The colony formation rate of colony formation unit megakaryocyte (CFU-MK) and burst formation unit-megakaryocyte (BFU-MK) were counted. RESULTS: There was no statistical difference on the positive rates of micromegakaryocyte and type I lymphoid small micromegakaryocyte between groups of ITP and control. The number of micromegakaryocyte and the formation rates of CFU MK in ITP group were significantly higher than those in control group. Among AA patients, the numbers of MK, micromegakaryocyte and the formation rates of CFU MK, BFU-MK in vitro significantly decreased. There was no significant difference in the positive rate of micromegakaryocyte between groups of MDS and control, but the number of micromegakaryocyte and the positive rate of type I lymphoid micromegakaryocyte were significantly higher than those of control group. There was no statistical difference of the formation rate of CFU-MK between groups of MDS and control. But in 63% childhood patients, the formation rate of CFU-MK decreased, 25% increased,and 13% was normal; BFU-MK formation rate decreased significantly in MDS group. CONCLUSION: Overproliferation of bone MKs may exist in most ITP patients. For obviating the nosogenetic factors, the normal MK releasing platelet could be easily found in the culture system. But the colony formation rate of MK decreased in a few patients with CITP. The abnormality of MK might be one of the reasons for thrombocytopenia in partial patients with ITP. Underproliferation of MKs may exist in AA, but no pathosishemogenesis was found. The dysfunction of early phase MK progenitor and stem cell might be the major reason for AA, but not the abnormality of hematopoietic microenvironment. There may be two kinds of megakaryocyte clones in bone marrow of children with MDS. One may be pathologic and potentially malignant micromegakaryocytes, the other may be the normal megakaryocytic precursors. The increase of pathologic MK resulted in abnormal development and maturation of MK in bone marrow. The change of megakaryopoiesis showed different in ITP, AA or MDS. Using bone marrow smear megakaryocyte counting, small micromegakaryocyte immunohistochemical detecting and the formation rate of bone marrow MK colony assay, the different thrombocytopenia can be diagnosed during the early stage of ITP, AA or MDS. PMID- 15833189 TI - [Drug eruption of erythema multiforme caused by amikacin in a case]. PMID- 15833190 TI - [Influence of mannitol on cerebral blood flow of post-resuscitation children as detected by transcranial Doppler ultrasound]. AB - OBJECTIVE: To detect the influence of mannitol on cerebral blood flow of post resuscitation children by transcranial Doppler (TCD). METHODS: The blood flow changes of left side middle cerebral artery (MCA) and extracranial internal carotid artery (EICA) were monitored by TCD in 21 post-resuscitation children. TCD waveforms, peak velocity (Vp), diastolic velocity (Vd), mean velocity (Vm) and pulsitility index (PI) of MCA and EICA were monitored daily and 30 minutes before and after the first dose of mannitol. Glasgow scores were estimated in the meantime. The patients were divided into 2 groups according to the patterns of diastolic flow. Patients in group I showed no diastolic flow or retrograde diastolic flow (n = 9), and patients in group II had positive diastolic flow (n = 12). RESULTS: Vp, Vd, Vm, PI of MCA and EICA in group I patients had no significant change after the administration of mannitol and the Glasgow scores were much lower than that of group II patients (P < 0.05). TCD waveforms showed no improvement in group I patients and their outcomes were poor. Vd, Vm and Vp of MCA and Vd of EICA in group II patients increased, PI of MCA decreased (P < 0.05), while Vp, Vm, PI of EICA had no significant change. TCD waveforms recovered within one week. All the patients in this group survived. CONCLUSIONS: The results suggested that mannitol could improve cerebral perfusion in patients with mild brain damage. These patients usually had increased diastolic blood flow in the early stage. Lack of or retrograde diastolic flow of TCD waveforms might be associated with severe brain damage with poor outcome. Mannitol had no effect on improving cerebral blood flow in these patients. PMID- 15833191 TI - [A case of congenital leukopenia]. PMID- 15833192 TI - [Early arterial atherosclerosis and level of plasma homocysteine in simply obese children]. AB - OBJECTIVE: To understand whether hyperhomocysteinemia and early arterial atherosclerosis exist in simply obese children. METHODS: Totally 68 simply obese children (age 6-14 years, mean 10.8 +/- 2.3 years) were enrolled in this study, 50 were male and 18 were female. Body mass index (BMI) of the obese children was equal to or more than 22. The height of the children was (145 +/- 22) cm. Meanwhile, 26 normal children (age 6 - 14 years, mean 10.9 +/- 2.0 years) were selected as control group, 17 of these children were male and 9 were female. Their height was (148.5 +/- 5.8) cm. There were no significant differences in height and age between the obese and the control children. The carotid intimal medial thickness (IMT), brachial artery flow-mediated vasodilation were examined by Doppler Flow/Dimension System and the liver was examined by B-mode ultrasound imager. Plasma homocysteine was determined by the automated chemiluminescent enzyme immunoassays. Serum lipid concentration was determined by biochemical analytic method. Blood pressure of the right upper limbs was measured. A detailed medical and family history was systematically recorded. RESULTS: BMI was (27.8 +/ 4.5) in the obese children and (16.2 +/- 2.5) in the controls. There was significant difference between two groups (P < 0.01). The obese children had significantly increased values than the controls for the carotid intimal-medial thickness (P < 0.01). Right carotid IMT, right inner-carotid IMT, left carotid IMT and left inner-carotid IMT were respectively (0.54 +/- 0.13) mm, (0.69 +/- 0.14) mm, (0.52 +/- 0.12) mm and (0.67 +/- 0.14) mm in obese children and were respectively (0.45 +/- 0.04) mm, (0.46 +/- 0.04) mm, (0.45 +/- 0.05) mm and (0.46 +/- 0.03) mm in control groups. Conversely, the flow-mediated brachial artery dilation of the obese children was significantly lower than that of the controls [(11.0 +/- 4.3)% vs. (17.5 +/- 4.9)%, P < 0.01]. The obese children had higher level of plasma homocysteine than the controls [(7.9 +/- 2.7) micromol/L vs. (5.6 +/- 2.1) micromol/L, P < 0.01]. Total cholesterol (TC) in the obese children dramatically increased, so did triglyceride concentration (TG), LDL-cholesterol (LDL-ch) and apolipoprotein-B (apo-B). Of the obese children, had fatty liver or the tendency to fatty liver. Six cases of the 68 obese children (8%) had hypertension. Of the 68 obese children, 57 (84%) had the history of consuming excessive food or taking less exercise. Forty-four percent of the obese children (30/68) came from the obese families in which at least one of the parents or grandparents was obese. Twenty-nine percent (20/68) and 22% (15/68) of the obese children respectively came from the families in which at least one of the parents or grandparents suffered from hypertension or coronary heart disease. CONCLUSION: Early arterial atherosclerotic changes existed in simply obese children. Hyperhomocysteinemia may be an important factor of the obesity-induced early arterial atherosclerosis during childhood. PMID- 15833193 TI - [Demonstration of a mechanism of anti-inflammatory effect of erythromycin on allergic airway inflammation in rat]. AB - OBJECTIVE: To investigate whether erythromycin exerts anti-inflammatory effect on allergic airway inflammation and whether erythromycin modulate allergic airway inflammation by inhibiting nuclear factor kappa B (NF-kappa B) activation. METHODS: Ovalbumin (OVA) together with aluminum hydroxide and Bordetella Pertussis was injected intraperitoneally to immunize SD rats and two weeks later 1% OVA was inhaled to challenge them for consecutive 7 days to mimic allergic airway inflammation. In treatment group, erythromycin was given orally (180 mg/kg.d) during the course of allergen exposure. WBC counts in bronchoalveolar lavage fluid (BALF) and lung specimen analysis were used to describe lung tissue inflammation. The expression of NF-kappa B subunit p65 in cell nucleus of lung tissue was measured by immunohistochemistry and NF-kappa B binding activation in lung tissue by electrophoresis mobility shift assay (EMSA). RESULTS: Lung tissue specimen analysis indicated that the severity of allergic inflammation was reduced in treatment group. The number of total WBC in BALF (x 10(8)/L) (31 +/- 22) was lower than that in model group animals (66 +/- 28), P < 0.01. The number (x 10(3)/mm(2)) of cells with nuclear staining of NF-kappa B per square millimeter of submucosal region around large bronchus (1.4 +/- 0.4) was lower than that in model group animals (2.6 +/- 0.6), P < 0.01. NF-kappa B binding activity (32 +/- 14) was lower than that of model group (46 +/- 17), P < 0.05. CONCLUSION: Erythromycin had an obvious protective role in allergic airway inflammation. Erythromycin inhibited NF-kappa B transcriptional activity to exert anti-inflammatory effect. PMID- 15833194 TI - [Long-term effects of delayed hyperbaric oxygen therapy on hypoxic-ischemic brain injury in neonatal rats]. AB - OBJECTIVE: To evaluate the long-term effects of delayed hyperbaric oxygen (HBO) therapy on neonatal rats with hypoxic-ischemic brain injury (HIBD). METHOD: Postnatal 7 days newborn rats (n = 52) were randomly set to three groups: control (n = 18, sham operation), HIBD (n = 17), or HBO (n = 17). Pups in the HBO group were subjected to hyperbaric oxygen treatment with 2 atmosphaera absolutus, 5 x 30 min at a 24 h intervals since 48-72 h after the HIBD model. All the animals were tested for the spatial learning and memory ability in the Morris water maze from postnatal days 37 to 41. At day-42, rats were decapitated and the brains were analyzed for morphological and histological changes, including brain shapes and weights, survival neurons, percentage of AchE positive area and NOS positive neurons in hippocampal CA1 region. RESULTS: Rats in HBO and HIBD groups displayed significant morphological and histological damages, as well as severe spatial learning and memory disability. The average escape latency of Morris water maze in HBO group [(56 +/- 23) s] and HIBD group [(56 +/- 22) s] were longer than the control [(23 +/- 16) s] (P < 0.05). The swimming time in HBO group [(30 +/- 5) s] and HIBD group [(29 +/- 6) s] were shorter than the control [(51 +/- 5) s] (P < 0.05). The swimming length in HBO group [(572 +/- 92) cm] and HIBD group [(548 +/ 92) cm] were shorter than the control [(989 +/- 101) cm] (P < 0.05). The weight of left brains in HBO group [(598 +/- 46) mg] and HIBD group [(601 +/- 59) mg] were lighter than the control [(984 +/- 18) mg] (P < 0.05). The survival neurons of hippocamal CA1 region in HBO group [(97 +/- 27)/mm] and HIBD group [(100 +/- 27)/mm] were less than the control [(183 +/- 8)/mm] (P < 0.05). The percentage of AchE-positive fibers in HBO group [(18.4 +/- 2.2)%] and HIBD group [(18.5 +/- 2.2)%] were less than the control [(27.5 +/- 2.2)%,] (P < 0.05). NOS-positive neurons in HBO group [(21 +/- 5)/mm(2)] and HIBD group [(19 +/- 4)/mm(2)] were also less than the control [(34 +/- 6)/mm(2)] (P < 0.05). CONCLUSION: Delayed HBO therapy resulted in no protection against either HIBD-induced brain morphological and histological deficits or spatial learning and memory disability. PMID- 15833195 TI - [Effect of dendritic cells stimulated by different ways on cytotoxic activity of cord blood derived cytokine-induced killer cells and natural killer cells]. PMID- 15833196 TI - [Inhibitory effects of Par-4 antisense oligodeoxynucleotide on apoptosis of PC12 cell induced by glutamate mediated by Akt1 phosphorylation]. PMID- 15833197 TI - [Expression and methylation of CpG islands in the promoter of p15INK4B gene in children with acute myeloid leukemia]. PMID- 15833198 TI - [Detailed allelic loss mapping on 6q21 in acute lymphoblastic leukemia of children and its relation with clinical cases]. PMID- 15833200 TI - [Resistance of Mycoplasma pneumoniae to macrolides antibiotics]. PMID- 15833199 TI - [Effect of phenylalanine on genes expression of Rho GTPases: possible relevance to neuron damage induced by phenylalanine]. PMID- 15833201 TI - [Predictive value of vascular endothelial growth factor in metastasis of nephroblastoma]. PMID- 15833202 TI - [The levels and relationship of anticardiolipin antibody and anti-beta 2 glycoprotein I antibody in the serum of children with Kawasaki disease]. PMID- 15833203 TI - [The incidence of periventricular-intraventricular hemorrhage in preterm infants and its high-risk factors]. PMID- 15833204 TI - [Clinical study on atypical pathogens of lower respiratory tract infection in children]. PMID- 15833205 TI - [Application of fiberoptic bronchoscopy in the pediatric intensive care unit]. PMID- 15833206 TI - [Aplastic anemia transformed into acute myeloblastic leukemia M1 8 years later: a case report]. PMID- 15833207 TI - [Autoimmune hemolytic anemia occurred in a neonate born to the mother with Sjogren's syndrome]. PMID- 15833208 TI - [Late-appearing t(9;22) (q34;q11) translocation in acute lymphoblastic leukemia with dic(8;9) (p11;q11) chromosome in a case]. PMID- 15833209 TI - [A case of McCune-Albright syndrome combined with pituitary tumor]. PMID- 15833210 TI - [Updates on diagnosis and management of Kawasaki disease unresponsive to intravenous gamma globulin]. PMID- 15833211 TI - [Development and maturation of human megakaryocyte]. PMID- 15833212 TI - [Effects of cefdinir in pediatric infectious diseases]. PMID- 15833213 TI - [Recent advances in pathological studies of bronchial asthma]. PMID- 15833214 TI - [Granulocytic sarcoma secondary to acute lymphoblastic leukemia in a boy]. PMID- 15833217 TI - High-dose zafirlukast in emergency department provides small benefit in acute asthma. PMID- 15833216 TI - False-positive PSA associated with increased worry and fears. PMID- 15833218 TI - Popular diets equally effective for losing weight. PMID- 15833219 TI - Aspirin plus PPI safer than clopidogrel if there is history of GI bleeding. PMID- 15833220 TI - Clinical decision rules accurately predict stroke risk in atrial fibrillation. PMID- 15833221 TI - Acute otitis media: making sense of recent guidelines on antimicrobial treatment. AB - High-dose amoxicillin (80 to 90 mg/kg/d divided twice daily) remains the drug of choice for treatment of acute otitis media despite increasing antimicrobial resistance. For persistent or recurrent acute otitis media, guidelines recommend high-dose amoxicillin/clavulanate (90/6.4 mg/kg/d), cefdinir, cefprozil, cefpodoxime, cefuroxime, or ceftriaxone. Increasing the dose of amoxicillin does not cover infection with beta-lactamase-producing pathogens; add the beta lactamase inhibitor clavulanate to amoxicillin, or choose a cephalosporin with good activity against S pneumoniae and good beta-lactamase stability. Key factors for enhancing compliance are taste of suspension, dosing frequency, and duration of therapy. PMID- 15833222 TI - Meningococcal vaccine: New product, new recommendations. AB - The Centers for Disease Control and Prevention now recommends that all adolescents aged 11 to 12 years receive a quadrivalent, conjugate meningococcal vaccine (MCV-4). With time, universal administration of meningococcal vaccine for this age group will make moot the question of whether entering college freshmen should receive meningococcal vaccine. It should lead to a marked reduction in a potentially catastrophic disease and contribute to the continued decline in morbidity and mortality from bacterial meningitis in the United States, which has largely been due to advances in vaccine technology. PMID- 15833223 TI - Of mites and men: reference bias in narrative review articles: a systematic review. AB - BACKGROUND: Citations in scientific articles may tend to favor the views presented. We studied whether there is such reference bias in narrative review articles that discuss interventions against house dust mites for people with asthma. DESIGN: Systematic review of reviews identified in a Medline search that expressed an opinion about the clinical effects of physical or chemical intervention methods. MAIN OUTCOME MEASURE: Positive bias was judged to have occurred if the reference list contained a higher proportion of trial references with significant results than among all trials available to the authors (published 2 years or more prior to the review). RESULTS: Seventy reviews were included, of which 63 (90%) recommended physical interventions. Forty-six reviews had trial references, 4 of these only to chemical interventions. In the remaining 42 reviews, reference bias was detected (P=2 x 10-8). The most quoted trial had only 7 patients per group, its claimed significant result was probably erroneous, and it did not report a clinical outcome. Intervention recommendations were often based on nonrandomized evidence, and the most quoted nonrandomized controlled study had included only 10 patients per group but claimed very positive results. CONCLUSION: The narrative review articles were severely biased, and their positive intervention recommendations are at variance with the systematic Cochrane Review on this topic and a recent very large trial of physical intervention, both of which failed to find an effect. PMID- 15833224 TI - About levels of evidence. PMID- 15833225 TI - Unilateral rash on a baby girl. AB - An 11-month-old baby girl came to the clinic with a pruritic rash. The rash initially appeared in her popliteal fossa 2 weeks before the visit. The eruption extended to the right leg, arm, and flank the week before the visit, subsequently spreading to the contralateral flank. Three weeks before to the eruption's appearance, the patient had an upper respiratory infection with a dry nonproductive cough, which resolved spontaneously without antibiotics. The physical examination revealed a healthy-appearing infant girl with excoriated erythematous papules coalescing into plaques on her right flexural arm that continued to the axilla and down the right flank to the flexural aspect of her leg (Figure 1). Her left side was essentially free of any rash (Figure 2). No cervical or axillary lymphadenopathy was noted, and the remainder of her exam was normal. What is your diagnosis? How would you manage this condition? PMID- 15833226 TI - How patients' trust relates to their involvement in medical care. AB - OBJECTIVES: To examine the connection between patients' trust and their attitudes toward seeking care, participating in medical decision making, and adhering to treatment recommendations. METHODS: Data were collected from a national telephone survey of English-speaking adults (N=553) in 1999. Eligibility requirements were some type of public or private health care coverage and having seen a physician or other health professional at least twice in the past 2 years. Five questions on preferred role in medical care were asked. Trust in physicians and satisfaction with care were separately measured using validated scales. RESULTS: The most significant predictor of patients' preferred role in medical care is trust in the medical profession. Views also varied by sex, age, health, education, income, number of visits/years with physician, past dispute with a physician, and satisfaction with care, but many of these bivariate associations were no longer significant in multivariate regression models. Views varied slightly by trust in the specific physician. There were no racial differences. CONCLUSIONS: A strong connection exists between patients' preferred involvement in medical care and trust in the medical profession, but only a slight connection with trust in their own physician. Increased trust in physicians generally is associated with greater willingness to seek care, to follow recommendations of physicians, and to grant them decisional authority. Higher trust in a specific physician is strongly associated only with greater reported adherence. Although higher trust in the medical profession appears to entail a more deferential role by patients, higher trust is also consistent with more active patient roles such as seeking care and adhering to treatment regimens. PMID- 15833227 TI - Vesiculobullous disease. PMID- 15833228 TI - Tramadol addiction. PMID- 15833229 TI - Treating community-acquired bacterial respiratory tract infections: update on etiology, diagnosis, and antimicrobial therapy. PMID- 15833230 TI - Clinical inquiries. What interventions reduce the risk of contrast nephropathy for high-risk patients? AB - Several interventions may reduce the risk of contrast nephropathy for high-risk patients; however, most evidence uses surrogate markers for clinically relevant outcomes. Because dehydration is a risk factor for developing contrast nephropathy, periprocedural hydration is routinely recommended (strength of recommendation [SOR]: C, expert opinion). Single studies have suggested that isotonic saline is associated with less risk than half-normal saline, and hydration with fluids containing sodium bicarbonate is more efficacious than those containing isotonic saline (SOR: B, single randomized controlled trial [RCT]). Oral acetylcysteine lowers the risk of postcontrast elevations in creatinine if taken more than 24 hours before contrast administration (SOR: A, RCTs). Acetylcysteine's low cost and favorable side effect profile make it an appealing option. Hypo-osmolar contrast media are less likely to induce contrast nephropathy than hyper-osmolar media (SOR: A, RCTs). Finally, hemofiltration might be considered for patients with extremely high risk of developing contrast nephropathy (SOR: B, single RCT). PMID- 15833231 TI - Clinical inquiries. What interventions can help patients stop using chewing tobacco? AB - Nicotine replacement therapy (NRT), including gum and patches, decreases cravings and short-term abstinence rates, but does not improve long-term abstinence (strength of recommendation [SOR]: B, meta-analysis of small randomized controlled studies [RCT]). It is unclear if bupropion has an effect on cessation rates (SOR: B, small RCTs with conflicting results). Behavioral interventions increase abstinence rates for smokeless tobacco users (SOR: B, meta-analysis of small RCTs). PMID- 15833232 TI - Clinical inquiries. Does furosemide decrease morbidity or mortality for patients with diastolic or systolic dysfunction? AB - No large-scale randomized, placebo-controlled trials evaluate furosemide's effect on mortality and long-term morbidity in diastolic or systolic dysfunction. In short-term studies, furosemide reduces edema, reduces hospitalizations, and improves exercise capacity in the setting of systolic dysfunction (strength of recommendation [SOR]: B, based upon low-quality randomized controlled trials). Furosemide and other diuretics reduce symptomatic volume overload in diastolic and systolic dysfunction (SOR: C, based on expert opinion). There is potential morbidity with the use of high-dose loop diuretics (volume contraction, electrolyte disturbances, and neuroendocrine activation). Use of high-dose loop diuretics for systolic dysfunction is associated with increased mortality, sudden death, and pump failure death (SOR: B, based on retrospective analyses of large scale randomized controlled trials). However, diuretic resistance or disease severity may explain these latter findings. PMID- 15833233 TI - Clinical inquiries. What is the best treatment for gastroesophageal reflux and vomiting in infants? AB - The literature on pediatric reflux can be divided into studies addressing clinically apparent reflux (vomiting or regurgitation) and reflux as measured by pH probe or other methods. Sodium alginate reduces vomiting and improves parents' assessment of symptoms (strength of recommendation [SOR]: B, small randomized controlled trial [RCT]). Formula thickened with rice cereal decreases the number of postprandial emesis episodes in infants with gastroesophageal reflux disease (GERD) (SOR: B, small RCT). There are conflicting data on the effect of carob bean gum as a formula thickener and its effect on regurgitation frequency (SOR: B, small RCTs). Metoclopramide does not affect vomiting or regurgitation, but is associated with greater weight gain in infants over 3 months with reflux (SOR: B, low-quality RCTs). Carob bean gum used as a formula thickener decreases reflux as measured by intraluminal impedance but not as measured by pH probe (SOR: B, RCT). Omeprazole and metoclopramide each improve the reflux index as measured by esophageal pH probe (SOR: B, RCT). Evidence is conflicting for other commonly used conservative measures (such as positional changes) or other medications for symptomatic relief of infant GERD. There is very limited evidence or expert opinion regarding breastfed infants, particularly with regard to preservation of breastfeeding during therapy. PMID- 15833234 TI - Clinical inquiries. Does anticoagulation prevent thrombosis for persons with fractures distal to the hip? AB - Low-molecular-weight heparin (LMWH) prophylaxis significantly reduces the total incidence of deep venous thrombosis (DVT) for patients with lower-limb fractures managed with surgical fixation and cast immobilization (strength of recommendation [SOR]: A, based on multiple randomized controlled studies [RCTs]). Evidence is insufficient to show whether LMWH specifically reduces the risk of clinically significant DVTs, and recommendations on its use are conflicting (SOR: C, based on expert opinion). Evidence is insufficient to recommend for or against warfarin prophylaxis for DVT in fractures distal to the hip (SOR: C, based on expert opinion). PMID- 15833235 TI - Clinical inquiries. How does tissue adhesive compare with suturing for superficial lacerations? AB - Tissue adhesives are effective and yield results comparable to those with conventional suturing of superficial, linear, and low-tension lacerations. The cosmetic outcome is similar; wound complications, such as infection and dehiscence, may be lower with tissue adhesives. Wound closure of superficial lacerations by tissue adhesives is quicker and less painful compared with conventional suturing (strength of recommendation: A, systematic reviews of randomized trials). PMID- 15833236 TI - Professional obligation. PMID- 15833237 TI - Preventive measures to reduce bioaerosol exposure during refuse collection: results of field studies in the real-life situation. AB - The aim of the present research project was to investigate and evaluate technical and organisational measures targeted at bioaerosol reduction during the refuse collection and to determine the exposure of refuse collectors to dust, fungi and endotoxins in an on-the-job situation with different vehicle technologies and states of hygiene. The following technical factors were found to influence the bioaerosol concentration at the refuse collector's workplace: compaction method, lifting device control, rave rail height, hopper depth, design of intake area and dust interception. For instance, notably higher total fungal counts were recorded with rotating drum compaction than with packer plate compaction. A hinged lid closure at the lifting device in conjunction with a suction unit induced a positive effect. In addition, the automatic lifting system had a positive influence on rear-end loaders, as did loading operation control from the closed driver's cab on side loaders. Regular internal and external high-pressure cleaning of the lifting device at intervals of not more than 14 days is recommended as a basic rule for vehicle hygiene. PMID- 15833238 TI - The development of a spatially explicit model to estimate radiocaesium body burdens in raccoons (Procyon lotor) for ecological risk assessment. AB - A spatially explicit model of raccoon (Procyon lotor) distribution for the U.S. Department of Energy's (DOE) Savannah River Site (SRS) in west-central South Carolina was developed using data from a raccoon radio-telemetry study and visualized within a Geographic Information System (GIS). An inductive approach was employed to develop three sub-models using the ecological requirements of raccoons studied in the following habitats: (1) man-made reservoirs, (2) bottomland hardwood/riverine systems, and (3) isolated wetland systems. Logistic regression was used to derive probabilistic resource selection functions using habitat compositional data and landscape metrics. The final distribution model provides a spatially explicit probability (likelihood of being in an area) surface for male raccoons. The model is a stand-alone tool consisting of algorithms independent of the specific GIS data layers to which they were derived. The model was then used to predict contaminant burdens in raccoons inhabiting a riverine system contaminated with radiocaesium (137Cs). The predicted 137Cs burdens were less than if one would assume homogeneous use of the contaminated areas. This modelling effort provides a template for DOE managed lands and other large government facilities to establish a framework for site specific ecological assessments that use wildlife species as endpoints. PMID- 15833239 TI - Isolation, molecular characterization and antimicrobial resistance patterns of Salmonella and Escherichia coli isolates from meat-based fast food in Lebanon. AB - The aim of this study was to characterize at the molecular level the different stains of Salmonella spp. and Escherichia coli that were isolated from meat-based fast food in Lebanon. In addition, this study evaluated the resistance of those strains to different antimicrobials that are commonly used. The foods included were Lahm-bi-Ajeen (LBA, meat pies) and Shawarma (Lebanese meat sandwiches similar to Gyros and Donairs, containing meat, vegetables, and sesame seeds-oil based sauce). Polymerase chain reaction (PCR) was used to characterize and identify the strains of both bacteria. Salmonella species characterization was performed using rfb genes cluster genetic marker, while that of E. coli strains were carried out based on stx1, stx2, eaeA, fliC, and ehlyA virulence markers. The characterized strains were then tested for their response to various antimicrobials. The results showed that the tested foods were contaminated with Salmonella paratyphi (serogroup A) and Shiga Toxin (Stx)-producing E. coli (STX EC). The PCR showed that 75% of E. coli tested strains was positive in PCR performed with stx1 primers, one of which was eaeA positive. Two of the tested strains were positive using PCR with fliC primers. The resistances of the various strains were evaluated using the following antimicrobials: Oxacillin, Teicoplanin, Trimethoprim/sulfamethoxazole, Gentamicin, Clindamycin, Cefotaxime, Cefuroxime, Erythromycin, and Vancomycin. Bacteria were highly resistant to one or more of the tested antimicrobials. Approximately 69% of E. coli and 77.8% of Salmonella spp. exhibited resistance. Salmonella spp. were shown to be 100% resistant to four antimicrobials: Oxacillin, Teicoplanin, Clindamycin, Vancomycin, and Erythromycin, while E. coli was 100% resistant to Teicoplanin and Trimethoprim/sulfamethoxazole. The most interesting findings were the high susceptibility of the E. coli to Gentamicin (100%). Highest resistance in the case of Salmonella spp. was seen against Cefotaxime (74%). Those two antimicrobials are commonly used for the treatment of enteric infections caused by gram-negative bacteria. The results showed that meat-based fast foods in Lebanon could be a public health hazard, especially Shawarma, as they may act as a potential vehicle for many antimicrobial-resistant pathogenic organisms. Improper hygienic standards and indiscriminate use of antimicrobials are two of the main causes for the prevalence of these pathogenic resistance strains in Lebanon. These results will emphasize the need to implement protective measures and more emphasis will be placed on the application of hygienic practices to reduce the levels of food contamination. PMID- 15833240 TI - Methyl mercury exposure in Swedish women with high fish consumption. AB - We studied the exposure to methyl mercury (MeHg) in 127 Swedish women of childbearing age with high consumption of various types of fish, using total mercury (T-Hg) in hair and MeHg in blood as biomarkers. Fish consumption was assessed using a food frequency questionnaire (FFQ), including detailed information about consumption of different fish species, reflecting average intake during 1 year. We also determined inorganic mercury (I-Hg) in blood, and selenium (Se) in serum. The average total fish consumption, as reported in the food frequency questionnaire, was approximately 4 times/week (range 1.6-19 times/week). Fish species potentially high in MeHg, included in the Swedish dietary advisories, was consumed by 79% of the women. About 10% consumed such species more than once a week, i.e., more than what is recommended. Other fish species potentially high in MeHg, not included in the Swedish dietary advisories, was consumed by 54% of the women. Eleven percent never consumed fish species potentially high in MeHg. T-Hg in hair (median 0.70 mg/kg; range 0.08-6.6 mg/kg) was associated with MeHg in blood (median 1.7 microg/L; range 0.30-14 microg/L; rs = 0.78; p < 0.001). Hair T-Hg, blood MeHg and serum Se (median 70 microg/L; range 46-154 microg/L) increased with increasing total fish consumption (rs = 0.32; p < 0.001, rs = 0.37; p < 0.001 and rs = 0.35; p = 0.002, respectively). I Hg in blood (median 0.24 microg/L; range 0.01-1.6 microg/L) increased with increasing number of dental amalgam fillings. We found no statistical significant associations between the various mercury species measured and the Se concentration in serum. Hair mercury levels exceeded the levels corresponding to the EPA reference dose (RfD) of 0.1 microg MeHg/kg b.w. per day in 20% of the women. Thus, there seems to be no margin of safety for neurodevelopmental effects in fetus, for women with high fish consumption unless they decrease their intake of certain fish species. PMID- 15833241 TI - Concentrations of lead and mercury in multimedia samples from homes near the former Clark Air Base, Philippines. AB - We measured lead and mercury in samples collected from 31 homes in communities near the former Clark Air Base, Philippines during May and October 2002. Sample media included water used for drinking and cooking, house dust and entryway soil. Composite samples of 15 food items purchased at local markets were also collected. Samples were analyzed for total lead (Pb) and total mercury (Hg) to evaluate the relative importance of each media to residential exposure concentrations in the community adjacent to Clark (Community A) versus a control community 5 km away (Community B). In general, we measured low (e.g. background) to undetectable levels of the target analytes in all media sampled with two important exceptions. First, the Hg concentrations we measured in canned mackerel composites, which were within the range reported for mackerel from other locations worldwide, may pose a risk to pregnant women who are frequent consumers (e.g. one or more cans per day). Second, we measured Pb above the USEPA residential screening concentration (400 mug/g) in dust and soil from two homes, illustrating the need for periodic residential lead monitoring in these and other communities in the Philippines. We found no significant difference between Communities A and B with respect to Pb and Hg concentrations in water or food, although we were not able to detect very low levels of Pb in most of the foods we sampled because of trace Pb contamination added during sample homogenization. Although the Pb levels we measured in dust and soil from Community A homes were higher on average than Community B homes, the levels in both communities were low (e.g. background) thus we did not investigate the difference further. To our knowledge, these are the first reported measurements of Pb in house dust in the Philippines. The concentrations of Pb we measured in house dust were significantly higher than those in entryway soil from both communities, adding empirical support to the assertion that yard soil should not be considered a proxy for house dust in exposure studies in the Philippines or elsewhere. PMID- 15833242 TI - Plasma vitellogenin levels and gonadal morphology of wild carp (Cyprinus carpio L.) in a receiving rivers downstream of sewage treatment plants. AB - A field study was carried out in order to assess, in a wild cyprinid fish, the biological effects related to the exposure to estrogenic compounds. The gonadosomic index (GSI), plasma vitellogenin (VTG) level and histological observations of gonads were performed to evaluate the reproductive health in common carp (Cyprinus carpio) from three rivers receiving sewage treatment plant (STP) effluents (Guadarrama, Henares and Jarama rivers, Spain) and from a reference site (Lozoya river). No significative differences in male GSI between sites were described. Only in the 18% of fish from Guadarrama river were elevated plasma VTG levels detected. The presence of oocytes in the testis was not found in any of males examined. The results suggest that the released estrogenic compounds by STPs effluents are not enough capable to adversely affect the gonadal development of the fish studied. Additional studies will be required to assess the risk of estrogenic compounds on fish from other sites of this area. PMID- 15833243 TI - Trends in fluctuating asymmetry in East Greenland polar bears (Ursus maritimus) from 1892 to 2002 in relation to organohalogen pollution. AB - Fluctuating asymmetry (FA) was studied in skulls of 283 polar bears (Ursus maritimus) sampled in East Greenland from 1892 to 2002. Fourteen metric bilateral traits in skull and lower jaw were measured and compared between polar bears born until 1960 (n = 94) and from 1961 (n = 189). The period 1892-1960 was chosen to represent a period prior to appearance of organohalogens (polychlorinatedbiphenyls [PCBs], dichlorodiphenyltrichloroethanes [DDTs], hexacyclohexanes [HCHs], chlordanes [CHLs], hexachlorobenzene [HCB], polybrominateddiphenylethers [PBDEs] and dieldrin) originating from long-range transport to East Greenland from southern latitudes. The period 1961-2002 represents the period when polar bears have been exposed to organohalogens. During this latter period, the level of organochlorines is believed to have increased from 1960 to the late 1980s followed by a likely decrease from 1990 to 2002. Within this later period, other compounds such as, e.g., polybrominated flame retardants are believed to have increased throughout the period. Two different analysis showed, that the degree of fluctuating asymmetry did not differ statistically between the two periods in ten of thirteen traits (one trait excluded due to high measurement error). In fact, when significant differences were found in four of the traits, the fluctuating asymmetry was lower in skulls sampled after 1960. The degree of fluctuating asymmetry was higher in adults than in subadults for 6 of the 14 traits, whereas a higher degree of fluctuating asymmetry was found for only one trait in one of the analyses for subadults relative to adults. Females had a higher degree of fluctuating asymmetry than males in one trait. A time trend analysis did find fluctuations over time for five traits but the relationship was weak as the trend appeared to occur by chance due to the high number of regressions analysed (n = 42). A correlation analysis of FA versus the sum concentrations of various classes of organohalogens in adipose tissue from a subsample of 94 recently collected polar bears (1999 2002) did not show a trend either. Hence, the present study could not document a relationship between skull asymmetry in polar bears and periods with different exposure to organohalogens. These findings are possibly influenced by nutritional status, genetic factors, a subeffect exposure of organohalogens or confounded by other environmental factors (e.g. temperature) within the two investigated periods. PMID- 15833244 TI - A comparison of native and transplanted Fontinalis antipyretica Hedw. as biomonitors of water polluted with heavy metals. AB - Native and transplanted mosses of the species Fontinalis antipyretica were studied to assess their capacity as biomonitors of heavy metals. Assays were carried out with transplanted mosses (sampled from an unpolluted control stream) exposed for 60 days to five streams polluted with heavy metals. At the same time, native mosses were collected from the exposure sites. Concentrations of N, P, K, Ca, Mg, S, Fe, Al, Ba, Cd, Co, Cr, Cu, Mn, Ni, Pb, V, and Zn were determined in the mosses (native and transplants), stream waters, and sediments of both exposure and control sites. The results showed that the transplanted mosses accumulated significantly more Al, Cr, Cu, Pb, V, and Zn than the native mosses. The concentrations of Co and Mn in all streams were significantly higher in the native mosses. PMID- 15833245 TI - Differential sensitivity of three cyanobacterial and five green algal species to organotins and pyrethroids pesticides. AB - In this work, five organotins and pyrethroids pesticides were tested to examine their effects on the three cyanobacteria Anabaena flos-aquae, Microcystis flos aquae, Mirocystis aeruginosa and on the five green algae Selenastrum capricornutun, Scenedesmus quadricauda, Scenedesmus obliqnus, Chlorella vulgaris, Chlorella pyrenoidosa through 96 h acute toxicity tests. The results indicated that: (1) the decreasing order of the average acute toxicity to cyanobacteria and green algae of five dissimilar organotins and pyrethroids pesticides was: fentin hydroxide > cyhexatin > azocyclotin > fenbutatin oxide > beta-cyfluthrin. (2) Wide variations occurred in response to the tested pesticides among the eight individual species of cyanobacteria and green algae. The sensitivity of various species of algae exposed to fenbutatin-oxide varied over one order of magnitude, exposed to cyhexatin/fentin-hydroxide/beta-cyfluthrin varied over two orders of magnitude and exposed to azocyclotin varied three orders of magnitude. (3) In contrast with the sensitivity of cyanobacteria and green algae, cyanobacteria were much less sensitive to beta-cyfluthrin than green algae. The pollutants may result in a shift of green algal and cyanobacterial group structure, especially in a shift from dominance by green algae to dominance by cyanobacteria, and may sustain cyanobcterial blooms during the special period. Thus, the decreasing order of the aquatic ecological risk was: beta-cyfluthrin > fentin hydroxide > cyhexatin > azocyclotin > fenbutatin oxide. There was a strong variance between toxicity and ecological risk, i.e. "low toxicity" does not automatically imply "low ecological risk". The toxicity of pyrethroids pesticides was lower than that of organotins pesticides, whereas the aquatic ecological risk of pyrethroids pesticides was higher than that of organotins pesticides. PMID- 15833246 TI - Histology of selected immunological organs in polar bear (Ursus maritimus) from East Greenland in relation to concentrations of organohalogen contaminants. AB - Samples of lymph nodes (axillary, n = 54 and inguinal, n = 45), spleen (n = 60), thymus (n = 11) and thyroid tissue (n = 5) from a total of 82 polar bears (Ursus maritimus) collected in East Greenland 1999-2002 were examined histologically. The purpose was to relate histology to concentrations of organohalogen contaminants (OHCs) [i.e., sum (Sigma)PCBs, SigmaDDTs, SigmaHCHs, SigmaCHLs, HCB, Dieldrin and SigmaPBDEs] determined in adipose tissue, as studies on polar bears have indicated that some OHCs act as immunosuppressive agents. Secondary follicle counts were evaluated in spleen and lymph nodes, and semiquantitively divided into four groups (0: few/absent to 3: high). In the spleen, a high secondary follicle count was found in 21% of the cases (12/60), and this was significantly higher in subadults (p < 0.01) compared to adults of both sexes. Also in the lymph nodes a high secondary follicle count was found in 20% of the cases (20/99), and in the axillary lymph nodes changes were significantly higher in subadults (p < 0.05) compared to adults of both sexes. Significantly weak correlations between concentrations of OHCs and the amount of secondary follicles in lymph nodes was found, but probably occurred as a consequence of other multiple stress factor(s) (all: p < 0.05) and also one significant, but modest positive correlation was found between SigmaPBDE concentrations and secondary follicle counts (p < 0.01; r = 0.41). In spleen, a significant relation between low concentrations of OHCs in adipose tissue and few/absent secondary follicles was found with respect to SigmaCHLs, SigmaHCHs, HCB and Dieldrin. No histopathological observations (e.g., neoplasia) were found in spleen or lymph nodes, nor in thymus or thyroid. In conclusion, the present data suggest that the exposure concentrations of OHCs to polar bears are unlikely to have resulted in adverse effects on the tissues in question, although SigmaCHLs, SigmaHCHs, HCB and Dieldrin were related to increased secondary follicle counts in the spleen. PMID- 15833247 TI - Trends in aerosol optical depth in the Russian Arctic and their links with synoptic climatology. AB - Temporal and spatial variability of aerosol optical depth (AOD) are examined using observations of direct solar radiation in the Eurasian Arctic for 1940 1990. AOD is estimated using empirical methods for 14 stations located between 66.2 degrees N and 80.6 degrees N, from the Kara Sea to the Chukchi Sea. While AOD exhibits a well-known springtime maximum and summertime minimum at all stations, atmospheric turbidity is higher in spring in the western (Kara-Laptev) part of the Eurasian Arctic. Between June and August, the eastern (East Siberian Chukchi) sector experiences higher transparency than the western part. A statistically significant positive trend in AOD was observed in the Kara-Laptev sector between the late 1950s and the early 1980s predominantly in spring when pollution-derived aerosol dominates the Arctic atmosphere but not in the eastern sector. Although all stations are remote, those with positive trends are located closer to the anthropogenic sources of air pollution. By contrast, a widespread decline in AOD was observed between 1982 and 1990 in the eastern Arctic in spring but was limited to two sites in the western Arctic. These results suggest that the post-1982 decline in anthropogenic emissions in Europe and the former Soviet Union has had a limited effect on aerosol load in the Arctic. The post-1982 negative trends in AOD in summer, when marine aerosol is present in the atmosphere, were more common in the west. The relationships between AOD and atmospheric circulation are examined using a synoptic climatology approach. In spring, AOD depends primarily on the strength and direction of air flow. Thus strong westerly and northerly flows result in low AOD values in the East Siberian Chukchi sector. By contrast, strong southerly flow associated with the passage of depressions results in high AOD in the Kara-Laptev sector and trajectory analysis points to the contribution of industrial regions of the sub-Arctic. In summer, low pressure gradient or anticyclonic conditions result in high atmospheric turbidity. The frequency of this weather type has declined significantly since the early 1980s in the Kara-Laptev sector, which partly explains the decline in summer AOD values. PMID- 15833248 TI - Seasonal and long-term change in lead deposition in central Japan: evidence for atmospheric transport from continental Asia. AB - Long-range transport of air pollution from continental Asia is currently an important issue concerning the Japanese environment, especially in regions susceptible to acidification due to low buffering capacity, such as Murakami, Niigata prefecture, located on the west coast of central Japan. Evidence for long range transport was obtained through lead and lead isotopic analysis of 84 archived precipitation filters, showing seasonal changes in lead deposition from May 1999 to May 2002. Lead deposition was highest in winter and spring (November through May) each year and lowest in summer. Computed 72-h back trajectories showed that in winter air masses were predominantly transported from the northwest, passing over northern China and eastern Russia, whilst in summer air masses predominantly originated from the southeast passing over Japan. Lead isotopic analysis showed higher (208)Pb/(206)Pb during winter, indicating that lead originated from a different source. A plot of (207)Pb/(206)Pb vs. (208)Pb/(206)Pb identified a thorogenic component, which is excess (208)Pb compared to a standard lead growth curve, indicative of certain lead ores and coals in continental Asia. The data provided evidence of long-range transport of lead from continental Asia to Japan. Bark pockets included within the trunks of two Japanese cedar trees harvested near Murakami, dating between 1972 and 1982, exhibited lead isotope ratios indicative of Japanese-sourced lead. In contrast, current (2003) bark showed thorogenic ratios, consistent with a relative decline in Japanese-sourced and increase in continental-sourced lead. PMID- 15833249 TI - Distribution of natural and anthropogenic thallium in the soils in an industrial pyrite slag disposing area. AB - The total concentrations combined with the chemical speciation of thallium (Tl) were examined in order to track the distribution of natural and anthropogenic Tl in the soils in an industrial pyrite slag disposing area. Their geochemical behaviors in the soils were further discussed. Soil samples were collected from three soil profiles adjacent to a large open-disposed pile of industrial Tl-rich pyrite slag, and from one soil profile in the background area. The results show that the soil contamination with Tl derived from slag (slag-Tl) is generally limited; slag-Tl was mainly accumulated in the upper part (< 16.5 cm) of the vicinal soils of the slag pile and shows large variation in concentration among different sampling sites. Basically, the soils surrounding the slag pile within 5 m are more Tl-contaminated than those under the slag pile and those far away from the slag pile. In respect of the concentrations of total Tl, the deeper soils of the studied profiles seem to be uncontaminated. However, the percentages of Tl in the easily reducible fraction indicate that these soils have been actually contaminated by slag-Tl. Natural Tl and anthropogenic Tl are distributed differently among the soil components in the studied soils. Natural Tl in the background soils is predominantly hosted in the residual fraction ( approximately 98%), while anthropogenic Tl was significantly incorporated into the more labile fractions of the soil (up to approximately 80%), especially in the acid extractable fractions and easily reducible fraction (up to approximately 30% and approximately 45%, respectively). Detailed analysis of speciation data of Tl suggests that despite being predominantly controlled by the degree of Tl pollution, the distribution of slag-Tl in the soils can be further affected by the general differences in soil properties. In this study, the order for preferential immobilization of anthropogenic Tl among major soil components can be roughly summarized as: Tl(III) carbonates and hydroxides > Mn oxide-hydroxides > Fe oxide-hydroxides > adsorption sites on the surface of soil, while the order can be significantly mediated by the pH conditions in the soils. The correlations between the fractions of Tl in the slag and in the soils indicate that the anthropogenic Tl in the soils in the studied slag disposing area should be mainly derived from the dissolved slag-Tl that was leached by rainwater rather than from the washed-out particles of slag. PMID- 15833250 TI - A sensitivity study for the visualisation of bacterial weathering of concrete and stone with computerised X-ray microtomography. AB - Geologists and engineers recently have adopted computerised X-ray microtomography (microCT), a radiological imaging technique, for geological and petrophysical applications such as the assessment of sediment characteristics, CT-measurement of compressibility and compaction, multiphase flow studies, measurement of bulk density and moisture content, and of porosity and permeability. This study focuses on another application: the monitoring of biological weathering of natural building stones and concrete. Microbial activity as a determining factor in the deterioration process of building materials has a major economic impact. Because of its non-destructive character, microCT could be the ideal monitoring technique. With this technique, three-dimensional (3D) images of the entire inner structure of the material can be obtained, together with quantitative data. In depth changes of porosity of concrete and stone specimens due to bacterial weathering were assessed in this work. Also, porosity was visualised based on 3D data with homemade software. Scanning electron microscopy (SEM) images provided additional information and supported conclusions drawn from the X-ray microCT data. Resolution improvement will make the study of petrophysical aspects of physical weathering and/or biological deterioration processes of natural building stones and concrete a promising subject for further microCT-application. PMID- 15833251 TI - The impact of volcanic gases from Miyake island on the chemical constituents in precipitation in the Tokyo metropolitan area. AB - The volcano on Miyake Island first erupted in July 2000 and continuous emission of volcanic gas from the collapsed caldera has been observed from the middle of August 2000. The large volcanic emission of SO2 had a strong influence on Tokyo metropolitan area, which is located approximately 150 km north of Miyake Island. We measured major ions in precipitation and dry deposition samples which had been collected at five sampling sites (Yokohama, Kashiwa, Fujisawa, Yokosuka, and Hachioji) in the Tokyo metropolitan area for 12 years since 1990. We have evaluated quantitatively the impact of the volcanic SO2 gas emitted from Miyake Island on the Tokyo metropolitan area by comparing depositional ionic constituents in the volcanic degassing period (from September 2000 to August 2001) with those in the normal period of the past 10 years (September 1990 to August 2000). nss-SO4(2-) concentrations in precipitation at the sampling sites in the Tokyo metropolitan area were 59.5-77.0 microeq/L during the degassing period, and 33.3-44.1 microeq/L during the normal period, respectively. The difference of nss-SO4(2-) concentrations between the two periods was statistically significant. In contrast, no significant differences were observed in the concentrations of the other major ions (NH4+, nss-Ca2+, Cl-, and NO3-) between the two periods. The impact of volcanic degassing from Miyake Island on the ionic concentrations in the precipitation of the Tokyo metropolitan area was seen only in the H+ and nss-SO4(2-) concentrations. The annual wet deposition amount of volcanic nss-SO4(2-) into the Tokyo metropolitan area has been quantitatively estimated. The annual wet deposition amounts were calculated as 701+/-277 Meq/year (22.4+/-8.9 kt SO2/year) on the total area of the Tokyo metropolitan area (14,000 km2). The wet deposition amount of nss-SO4(2-) corresponds to only 0.15% of the total annual amount of volcanic SO2 (15 Mt/year) emitted from Miyake Island from September 2000 to August 2001. This estimation is consistent with a previous study which reported that 0.3% of the atmospheric discharge of SO2 from the volcano in Mt. Etna in Sicily was deposited by precipitation to the surrounding area (1200 km2). The annual wet deposition amount of volcanic SO2 to the Tokyo metropolitan area was at the same order of the magnitude (37+/-15%) as the annual anthropogenic SO2 amount (60 kt/year) emitted from the Tokyo metropolitan area. PMID- 15833252 TI - Autocatalyst-derived platinum, palladium and rhodium (PGE) in infiltration basin and wetland sediments receiving urban runoff. AB - The emission of platinum group elements (PGE) from automobile catalytic converters has led to rapid increases in Pt, Pd and Rh concentrations in roadside media. Significant quantities of the PGE may enter and accumulate in fluvial systems via road runoff. This paper examines the occurrence and spatial distribution of autocatalyst-derived PGE in surface sediments of infiltration basin and wetland sediments receiving road-runoff in Perth, Western Australia. Samples were analysed by ICP-MS following microwave digestion and cation exchange. PGE concentrations ranged between 1.5-17.2 Rh, 5.4-61.2 Pd and 9.0 103.8 ng g(-1) Pt. The highest levels of PGE were generally found at basin topographic low points and these concentrations were found to be strongly related to the area of road surface drained and the traffic volume. PGE ratios in infiltration basin and wetland sediments were within the typical range of catalytic converter compositions. However, comparisons of PGE ratios between parent road dusts and infiltration basin sediments revealed a systematic shift in Pt/Pd ratios, suggesting that PGE fractionation can occur during transport through the drainage system and that a small portion of Pd in road dust may be solubilised under natural conditions. PMID- 15833253 TI - Correlations, partitioning and bioaccumulation of heavy metals between different compartments of Lake Balaton. AB - Correlations between trace metals in dissolved and particulate phases, zooplankton, mussels and sediments in Lake Balaton were investigated. The degree of correlation between the various metals was different in each of the investigated compartments. Particulate metal concentrations (microg g(-1)) were anti-correlated with suspended particulate matter (SPM) (mg l(-1)), indicating a dilution effect, while total metal concentrations in the water column (microg l( 1)) were highly correlated with SPM, implying a major influence of the turbidity on the total metal concentrations. Between compartments, not many significant correlations were recognized. Only Ba, Ca, Sr and Mg are correlated in the sediments and in the particulate phase, suggesting common sources for both compartments. Partitioning coefficients (Kd) of trace metals between dissolved and particulate phases are generally low, typical for natural water and fairly stable over the lake. Most of the trace metals (Zn, Co, Cd and Pb) exist in the particulate phase (for about 70% of the total metal load). Cu and Ni are exceptions, showing a more equal distribution. Bioconcentration factors (BCF) of zooplankton and mussels were comparable to those of other natural waters. A negative biomagnification from suspended particulate matter to zooplankton and from sediment to mussel was recognized for all trace metals, except a small enrichment of Zn in zooplankton and Cd in mussel. Four factors were recognized in SPM and in sediments but they did not contain the same group of metals. Cluster analysis showed that metal accumulations in the sediments were different between northern and southern shores and in SPM between western and eastern areas. PMID- 15833254 TI - Metal transfer to plants grown on a dredged sediment: use of radioactive isotope 203Hg and titanium. AB - Improperly disposed of dredged sediments contaminated with metals may induce long term leaching and an increase of metal concentrations in ground waters and vegetal cover plants. The objective of the study was to quantify the sediment-to plant transfer of Cu, Pb, Hg and Zn with a particular focus on the pathway of Hg and to determine whether the establishment of vegetal cover modifies the metal availability. A pot experiment with rape (Brassica napus), cabbage (Brassica oleraccea) and red fescue (Festuca rubra) was set up using a sediment first spiked with the radioisotope 203Hg. Zinc concentrations (197-543 mg kg(-1) DM) in leaves were higher than Cu concentration (197-543 mg kg(-1) DM), Pb concentration (2.3-2.6 mg kg(-1) DM) and Hg concentration (0.9-1.7 mg kg(-1) DM). Leaves-to sediment ratios decreased as follows: Zn > Cu > Hg > Pb. According to Ti measurements, metal contamination by dry deposition was less than 1%. Mercury concentration in plant leaves was higher than European and French thresholds. Foliar absorption of volatile Hg was a major pathway for Hg contamination with a root absorption of Hg higher in rape than in cabbage and red fescue. Growth of each species increased Cu solubility. Zinc solubility was increased only in the presence of rape. The highest increase of Cu solubility was observed for red fescue whereas this species largely decreased Zn solubility. Dissolved organic carbon (DOC) measurements suggested that Cu solubilisation could result from organic matter or release of natural plant exudates. Dissolved inorganic carbon (DIC) measures suggested that the high Zn solubility in the presence of rape could originate from a generation of acidity in rape rhizosphere and a subsequent dissolution of calcium carbonates. Consequently, emission of volatile Hg from contaminated dredged sediments and also the potential increase of metal solubility by a vegetal cover of grass when used in phytostabilisation must be taken into account by decision makers. PMID- 15833255 TI - Elemental composition of tropospheric aerosols in Hanoi, Vietnam and Nairobi, Kenya. AB - Air pollution problems in major cities within the developing countries need to be studied. There are scanty measurements from the developing countries on airborne particles despite their adverse implications to human health, visibility and climate. One of the major sources of anthropogenic air pollution is energy production. Energy demand is bound to increase as population increases, especially in major cities of the world. Fine particles, particles with aerodynamic diameter < or = 2.5 microm, are mainly anthropogenic and these particles were collected in the capital cities of Vietnam and Kenya. A cyclone airborne particle collector was used to sample in Hanoi during the months of May to October 2000 and a dichotomous virtual impactor in Nairobi in February 2000. The samples were analysed for elemental content by an energy dispersive X-ray fluorescence (EDXRF) spectrometer. S, Cl, K and Fe exceeded atmospheric concentrations of 100 ng m(-3) at both cities. Atmospheric elemental concentrations in both Hanoi and Nairobi were orders of magnitude higher than their respective rural towns. Traffic, biomass and waste burning emissions were implicated as the main sources of air pollution in Nairobi, while coal combustion and road transport were the major sources in Hanoi. Regional air pollution had a major impact over Hanoi, whereas an influence of that kind was not identified in Nairobi. Pb and other toxic elements had concentration levels below WHO guideline, however, the two cities are threatened by future high levels of air pollution due to the high rate of population growth. Long-term measurements are required in both areas to evaluate if the alarming situation is deteriorating. PMID- 15833256 TI - Differences in Cd and Zn bioaccumulation for the flood-tolerant Salix cinerea rooting in seasonally flooded contaminated sediments. AB - Several authors suggest that a hydrological regime aiming at wetland creation is a potential management option that favours reducing bioavailability for metal contaminated sites. The hydrological conditions on a site constitute one of the many factors that may affect the availability of potentially toxic trace metals for uptake by plants. Bioavailability of Cd, Mn and Zn on a contaminated dredged sediment landfill (DSL) with variable duration of submersion was evaluated by measuring metal concentrations in the wetland plant species Salix cinerea in field conditions. Longer submersion periods in the field caused lower Cd and Zn concentrations in the leaves in the first weeks of the growing season. Foliar Cd and Zn concentrations at the end of the growing season were highest on the initially flooded plot that emerged early in the growing season. Foliar Zn concentrations were also high at a sandy-textured oxic plot with low soil metal concentrations. Zn uptake in the leaves was markedly slower than Cd uptake for trees growing on soils with prolonged waterlogging during the growing season, pointing at a different availability. Zn availability was lowest when soil was submerged, but metal transfer from stems and twigs to leaves may mask the lower availability of Cd in submerged soils. Especially for Cd, a transfer effect from one growing season to the next season was observed: oxic conditions at the end of the previous growing season seem to determine at least partly the foliar concentrations for S. cinerea through this metal transfer mechanism. Duration of the submersion period is a key factor for bioavailability inasmuch as initially submerged soils emerging only in the second half of the growing season resulted in elevated Cd and Zn foliar concentrations at that time. PMID- 15833257 TI - Soil enzyme activities as affected by anthropogenic alterations: intensive agricultural practices and organic pollution. AB - The activity of a range of enzymes related to the cycling of the main biologically important nutrients C, N, P and S was investigated in cultivated and non-cultivated soils from various parts of Europe. Two agricultural sites from North Italy under continuous corn (Zea mays L.) with and without organic fertilization were compared. Two other agricultural sites from South Italy under hazel (Corylus avellana L.) never flooded or repeatedly flooded over by uncontrolled urban and industrial wastes were investigated. The non-cultivated soils were from Middle and South Europe with different pollution history such as no-pollution and pollution with organic contaminants, which is phenanthrene and other polycyclic aromatic hydrocarbons (PAHs). Agricultural soils showed significant differences in some of physical-chemical properties (i.e. organic C, total and labile phosphate contents, available Ca and Mg) between the two sites studied. Enzyme activities of hazel sites periodically flooded by wastes were mainly higher than in the hazel sites never flooded. Sites under many years of continuous corn showed dehydrogenase, invertase, arylsulphatase and beta glucosidase activities generally lower than the soils under hazel either flooded or not by wastes. As compared to agricultural soils, non-cultivated soils heavily or moderately polluted by organic contaminants displayed much lower values or complete absence of enzymatic activities. Dissimilar, contradictory correlations between soil enzyme activities and the majority of soil properties were observed separately in the two groups of soils. When the whole set of enzyme activities and soil properties were considered, all significant correlations found separately for the groups of soils were lost. The overall results seem to confirm that no direct cause-effect relationships can be derived between the changes of a soil in response to a given factor and both the variations of the activity and the behaviour of the enzymes in soil. PMID- 15833258 TI - [Is early treatment of infants at risk of spastic diplegia warranted?]. AB - OBJECTIVE: To test the possible link between first investigation and development of spastic diplegia. METHOD: A long-term retrospective study was carried out in the Neonatal Medicine Department of Arras Hospital, the C.A.M.S.P. of that town and with corresponding therapists. RESULTS: The study undertaken from January 1, 1991, to December 31, 1997, involved 56 children suspected of developing spastic diplegia and requiring a specialized course of treatment during the evolution of disease. In 32, evolution of disease was favourable, and in 24 spastic diplegia developed. An unfavourable evolution was significantly associated with late intervention of rehabilitation. CONCLUSION: Although few pre- or neonatal factors differentiated the two groups of children in their opposite evolution, haemorrhage during pregnancy and early detection seem to be determining factors in spastic diplegia. PMID- 15833259 TI - [Outcome of early fitting by silicone liner with lock system for dysvascular below-knee amputation: assessment of walking abilities by a specific questionnaire]. AB - PURPOSE: To assess by a specific questionnaire the functional outcome of patients with below-the-knee amputation after early prosthetic fitting by the ICEROSS silicone liner, which had demonstrated improvement of stump healing and length of hospital stay. PATIENTS AND METHODS: In this retrospective study, walking ability was assessed by a specific score resulting from answers on a questionnaire. The outcome variables were walking inside and outside, transfer from sitting, climbing stairs, and use of walking aids. Following amputation, the ICEROSS system was used for compression therapy, then for temporary prosthesis. The questionnaire was administered at the fitting stabilized state. RESULTS: Twenty nine of 51 patients who underwent trans-tibial amputation were included: 5 women (mean age 72.8+/-4.1 years) and 24 men (mean age: 69+/-7.4 years). The mean total score was 14.5/20 (good functional outcome) for the 22 unilateral amputees and 7.2/20 (intermediate result) for the seven bilateral amputees. Previous studies concerning functional outcome with other contact casts (without a silicon liner with a bolt) had shown similar results. CONCLUSION: Despite its beneficial initial effect, early fitting by the ICEROSS system did not improve walking ability at the steady functional state, which is more linked to advanced age and comorbidities. PMID- 15833260 TI - [Neuromuscular effects of superimposed and combined transcutaneous electrical stimulation with voluntary activity: a review]. AB - With voluntary muscular contraction (VOL), small motor units (MUs) are recruited before large MUs are (a submaximal muscular contraction recruits only small MUs), whereas electrical stimulation (ES) tends to reverse the recruitment order. On the basis of this observation, some authors have tested the physiological effects of ES superimposed simultaneously with VOL (superimposed technique [ST]) or separately (combined technique [CT]). With healthy subjects, ST does not recruit more MUs than VOL, except with eccentric contractions. After health subjects undergo training programs, ST appears to be as efficient as VOL in enhancing subjects' neuromuscular qualities. Nevertheless, the use of CT seems more effective than VOL. In postsurgical rehabilitation, both ST and CT are more effective than VOL. Actually, following knee surgery, ST and CT compensate for volume and muscle strength deficits with more efficiency than does VOL. PMID- 15833261 TI - [Incidence, circumstances and consequences of falls in patients undergoing rehabilitation after a first stroke]. AB - OBJECTIVES: To describe the incidence, circumstances, and consequences of falls in patients admitted in a rehabilitation ward after a stroke. METHODS: Prospective monitoring of falls over four years in a neurological rehabilitation unit. Use of fall registry. INCLUSION CRITERIA: age < 75 years, admission < 45 days after stroke onset, single stroke of 1 cerebral hemisphere (nonlacunar) or of the brain stem. Only falls due to loss of balance were considered; falls caused by a seizure or syncope were not considered. RESULTS: Of 217 consecutive patients with the inclusion criteria, 34 had fallen at least once (15.7%) and 10 twice (4.1%). Fall incidence, defined as the number of falls per patient per day was 2.2 per thousand. Half of the patients fell the first three weeks after admission. Most falls involved getting to or from the wheelchair or the bed; 1 patient had recovered minimal postural abilities at the gym but was not independent. Traumatic lesions were noted in 13 patients: they were minor in nine and severe in four, including three fractures. CONCLUSION: Falls due to loss of balance are a major problem in patients undergoing rehabilitation after a stroke. Getting to and from wheelchairs in the bedroom and bathroom by patients who are not allowed to do so play a key role in many falls. Prevention programs should consider this information. PMID- 15833262 TI - [Charcot-Marie-Tooth disease associated with syringomyelia revealed by urinary disorders]. AB - PURPOSE: Charcot-Marie-Tooth (CMT) disease is characterized by genetic and clinical heterogeneity. The occurrence of urinary disorders is unusual and requires further investigation. CMT disease and syringomyelia are not usually associated. Moreover, primary urinary signs in syringomyelia are not frequent. CASE REPORT: An 11-year-old girl with CMT disease 1A duplication presented with motor deficit of the leg associated with urinary disorders. Urodynamic study revealed neurogenic detrusor overactivity, and magnetic resonance imaging confirmed syringomyelia extending from T11 to an L1 located conus. DISCUSSION: Review of the literature confirmed the possibility of primary urinary signs in syringomyelia, with only one case of CMT and urinary signs described. In this case, the diagnosis was radicullar and medullar compression by a hypertrophic nerve formation called an "onion bulb" reflecting the demyelinisation and remyelinisation cycle. CONCLUSION: Urinary signs in CMT disease are exceptional and require investigation into the possibility of a central lesion. PMID- 15833263 TI - [What is the place of diphosphonates in the treatment of complex regional pain syndrome I?]. AB - OBJECTIVES: To evaluate the efficacy of biphosphonates (BPs) in complex regional pain syndrome I or reflex sympathetic dystrophy and to specify their place in this chronic painful syndrome. METHODS: A literature review of Medline and Embase with use of a combination of 3 key words: biphosphonates therapy, reflex sympathetic dystrophy, complex regional pain syndrome I, controlled study. Each article was classified by 2 independent reviewers according to Aguilar's method into high, middle or low quality. RESULTS: Three second-generation BPs were tested in the treatment of reflex sympathetic dystrophy: pamidronate, alendronate and clodronate. Many open studies investigated pamidronate, but only 2 were randomised and controlled: a low-quality trial comparing pamidronate with placebo and a middle-quality trial comparing pamidronate with calcitonine. Two high quality trials were performed, 1 with alendronate and 1 with clodronate. Middle quality trial and high-quality trials reported positive effects on pain with BPs in recent reflex sympathetic dystrophy (disease duration less than eight months). Side effects (fever, asymptomatic hypocalcemia) were observed frequently but disappeared quickly. CONCLUSION: BPs can be used in recent reflex sympathetic dystrophy when calcitonin is inefficient or when calcitonin injections are not well tolerated. PMID- 15833264 TI - [Which treatments for algodystrophies in 2005?]. PMID- 15833266 TI - Ovarian theca cell. AB - Theca cells are the endocrine cells associated with ovarian follicles that play an essential role in fertility by producing the androgen substrate required for ovarian estrogen biosynthesis. Theca cells differentiate from the interfollicular stroma in response to proteins secreted from growing follicles. The most common endocrine cause of infertility is associated with excessive proliferation of theca cells and ovarian hyperandrogenism. Cell facts: -ovarian androgen-producing cells; -are associated only with developing follicles; -over-activity of theca cells causes infertility due to hyperandrogenism; -under-activity of theca cells causes infertility due to lack of estrogen. Theca cells: androgen-producing cells in the ovary. PMID- 15833265 TI - Natural killer T cells: rapid responders controlling immunity and disease. AB - Natural killer T (NKT) cells are a subset of T cells that share properties of natural killer cells and conventional T cells. They are involved in immediate immune responses, tumor rejection, immune surveillance and control of autoimmune diseases. Most NKT cells express both an invariant T cell antigen receptor and the NK cell receptor NK1.1, and are referred to as invariant NKT cells. This invariant T cell receptor is restricted to interactions with glycolipids presented by the non-classical MHC, CD1d. These NKT cells rapidly produce high levels of interleukin (IL)-2, IFN-gamma, TNF-alpha, and IL-4 upon stimulation through their TCR. Most also have cytotoxic activity similar to NK cells. NKT cells are involved in a number of pathological conditions, and have been shown to regulate viral infections in vivo, and control tumor growth. They may also play both protective and harmful roles in the progression of certain autoimmune diseases, such as diabetes, lupus, atherosclerosis, and allergen-induced asthma. PMID- 15833267 TI - NaSi-1 and Sat-1: structure, function and transcriptional regulation of two genes encoding renal proximal tubular sulfate transporters. AB - Sulfate (SO(4)2-) is an important anion regulating many metabolic and cellular processes. Maintenance of SO4(2-) homeostasis occurs in the renal proximal tubule via membrane transport proteins. Two SO(4)2- transporters that have been characterized and implicated in regulating serum SO4(2-) levels are: NaSi-1, a Na+-SO(4)2- cotransporter located at the brush border membrane and Sat-1, a SO4(2 )-anion exchanger located on the basolateral membranes of proximal tubular cells. Unlike Sat-1, for which very few studies have looked at regulation of its expression, NaSi-1 has been shown to be regulated by various hormones and dietary conditions in vivo. To study this further, NaSi-1 (SLC13A1) and Sat-1 (SLC26A1) gene structures were determined and recent studies have characterized their respective gene promoters. This review presents the current understanding of the transcriptional regulation of NaSi-1 and Sat-1, and describes possible pathogenetic implications which arise as a consequence of altered SO(4)2- homeostasis. PMID- 15833268 TI - Working outside the system: an update on the unconventional behavior of the renin angiotensin system components. AB - The renin-angiotensin system (RAS) plays an important role in regulating arterial pressure, blood volume, thirst, cardiac function, and cellular growth. Both a circulating and multiple tissue-localized systems have been identified, and are generally portrayed as a series of reactions that occur sequentially with a single outcome: angiotensinogen is cleaved by renin to form angiotensin I, which in turn is processed by angiotensin-converting enzyme (ACE) to angiotensin II, which then activates either the AT1 or the AT2 plasma membrane receptor. Evidence has emerged, however, showing that some RAS components play important roles outside of this canonical scheme. This article provides an overview of some recently identified extra-system functions. In addition to forming angiotensin II, ACE is a multifunctional enzyme equally important in the metabolism of vasodilator and antifibrotic peptides. As the membrane-bound form, ACE functions as a "receptor" that initiates intracellular signaling leading to gene expression. Both angiotensin I and II may lead to actions that are independent of, or even oppose, those of the RAS via their metabolism by the novel ACE homologue ACE2. The two angiotensin II receptor types have ligand-independent roles that influence cellular signaling and growth, some of which may result from the ability to form hetero-dimers with other 7-transmembrane receptors. Finally, intracellular angiotensin II has been demonstrated to have actions on cell communication, gene expression, and cellular growth, through both receptor dependent and independent means. A greater understanding of these extra-system functions of the RAS components may aid in the development of novel treatments for hypertension, myocardial ischemia, and heart failure. PMID- 15833269 TI - Polychlorinated biphenyl (PCB) alters acid-sensitivity of cultured neurons derived from the medulla oblongata. AB - Polychlorinated biphenyls (PCBs) are known as environmental pollutants that may cause adverse health problems. However, little is known about the effects of PCBs on acid-sensitive neurons of the medulla oblongata, which regulate respiration. Therefore, the present study was designed to examine whether PCB alters acid sensitivity of cultured neurons derived from the rat medulla oblongata. When extracellular pH was shifted from 7.4 to 7.0, acid-sensitive neurons showed depolarization, which was measured by voltage-sensitive fluorescent dye. Exposure to PCB (Aroclor 1254) decreased the amplitude of depolarization in low pH and increased the resting membrane potential in a dose-dependent manner. Taken together, our results indicate that PCB potentially influences acid-sensitivity through alteration of the membrane potential of acid-sensitive neurons, which could affect the regulation of respiration. PMID- 15833270 TI - On the chaperonin activity of GroEL at heat-shock temperature. AB - The studies of GroEL, almost exclusively, have been concerned with the function of the chaperonin under non-stress conditions, and little is known about the role of GroEL during heat shock. Being a heat shock protein, GroEL deserves to be studied under heat shock temperature. As a model for heat shock in vitro, we have investigated the interaction of GroEL with the enzyme rhodanese undergoing thermal unfolding at 43 degrees C. GroEL interacted strongly with the unfolding enzyme forming a binary complex. Active rhodanese (82%) could be recovered by releasing the enzyme from GroEL after the addition of several components, e.g. ATP and the co-chaperonin GroES. After evaluating the stability of the GroEL rhodanese complex, as a function of the percentage of active rhodanese that could be released from GroEL with time, we found that the complex had a half-life of only one and half-hours at 43 degrees C; while, it remained stable at 25 degrees C for more than 2 weeks. Interestingly, the GroEL-rhodanese complex remained intact and only 13% of its ATPase activity was lost during its incubation at 43 degrees C. Further, rhodanese underwent a conformational change over time while it was bound to GroEL at 43 degrees C. Overall, our results indicated that the inability to recover active enzyme at 43 degrees C from the GroEL-rhodanese complex was not due to the disruption of the complex or aggregation of rhodanese, but rather to the partial loss of its ATPase activity and/or to the inability of rhodanese to be released from GroEL due to a conformational change. PMID- 15833271 TI - Folding and stability of a coiled-coil investigated using chemical and physical denaturing agents: comparative analysis of polymerized and non-polymerized forms of alpha-tropomyosin. AB - alpha-Tropomyosin (Tm) is a two-stranded alpha-helical coiled-coil protein, which participates in the regulation of muscle contraction. Unlike Tm purified from vertebrate muscle, recombinant Tm expressed in Escherichia coli is not acetylated at the N-terminal residue and loses the capacity to undergo head-to-tail polymerization, to bind actin and to inhibit actomyosin ATPase activity. These functions are restored by fusion of an N-terminal Ala-Ser (AS) dipeptide tail to recombinant Tm. Here, we have employed chemical (guanidine hydrochloride and urea) and physical (elevated hydrostatic pressures and low temperatures) denaturing agents to compare the structural stabilities of polymeric alanine serine-tropomyosin (ASTm, containing the AS dipeptide) and dimeric "non-fusion" Tm (nfTm, i.e., not containing the AS dipeptide). Binding of the hydrophobic fluorescent dye bis-ANS, circular dichroism and size-exclusion chromatography were used to monitor the stabilities and state of association of both proteins under different solution conditions. Bis-ANS binding was markedly decreased at low concentrations (<1M) of GdnHCl or urea, whereas the secondary structures of both ASTm and nfTm were essentially unaffected in the same range of denaturant concentrations. These results suggest local unfolding of bis-ANS binding domains prior to global unfolding of Tm. In contrast, increased bis-ANS binding was observed when Tm was submitted to high pressures or to low temperatures, implying increased exposure of hydrophobic domains in the protein. Taken together, the different sensitivities of ASTm and nfTm to different denaturing agents support the notion that, at close to physiological conditions, head-to-tail interactions in polymerized ASTm are predominantly stabilized by electrostatic interactions between adjacent Tm dimers, whereas non-polar interactions appear to play a major role in the stability of the coiled-coil structure of individual Tm dimers. PMID- 15833272 TI - The role of STAT-3 in the mediation of smooth muscle cell response to cyclic strain. AB - Hemodynamic forces, including shear stress and cyclic strain, have been recognised as important modulators of vascular cell morphology and function. However, the mechanism by which vascular cells sense and transduce the extracellular mechanical signals into the cell nucleus has not yet been clarified. The purpose of our study was to assess the involvement of the signal transducer and activator of transcription-3 (STAT-3) in the signaling pathway mediating the response of vascular smooth muscle cells (SMC) to cyclic strain. Embryonic A7r5 SMC derived from thoracic aortas of DB1X rats were seeded on flexible collagen I-coated plates. Cells were subjected to 10% average strain at 60 cycles/min for various time periods. Activation of STAT-3, p38, extracellular signal-regulated kinase (ERK) 1/2 and Src was assessed by immunoblotting using phosphospecific antibodies. The interactions between STAT-3 phosphorylation and p38, ERK1/2, phosphatidylinositol-3 (PI3K), mammalian target of rapamycin (mTOR), Janus kinase (JAK) 2 and Src were evaluated by pretreating the cells with specific inhibitors including SB202190, PD98059, LY294002, wortmannin, rapamycin, AG490 and PP1. Serine phosphorylation of STAT-3 was increased by 2-fold after 15 min of cyclic strain, while tyrosine phosphorylation was increased by 2.3-fold after 60 min. Inhibition of ERK1/2 by PD98059 prevented serine phosphorylation of STAT-3, whereas inhibition of Src by PP1 prevented STAT-3 tyrosine phosphorylation. Pretreating the cells with SB202190, a specific inhibitor of p38, resulted in an increase in basal phosphorylation of ERK1/2 and a subsequent increase in basal serine phosphorylation of STAT-3. In conclusion, both serine and tyrosine phosphorylation of STAT-3 are involved in the signaling pathway mediating the effects of cyclic strain on vascular SMC. Serine phosphorylation of STAT-3 is mediated by ERK1/2, while tyrosine phosphorylation is mediated by Src. A negative feedback loop was also found between p38 and ERK1/2. PMID- 15833273 TI - Irreversible cellular senescence induced by prolonged exposure to H2O2 involves DNA-damage-and-repair genes and telomere shortening. AB - H2O2 has been the most commonly used inducer for stress-induced premature senescence (SIPS), which shares features of replicative senescence. However, there is still uncertainty whether SIPS and replicative senescence differ or utilize different pathways. 'Young' human diploid fibroblasts (HDFs), treated with prolonged low doses of hydrogen peroxide, led to irreversible cellular senescence. Cells exhibited senescent-morphological features, irreversible G1 cell cycle arrest and irreversible senescence-associated beta-galactosidase positivity. The appearance of these cellular senescence markers was accompanied by significant increases of p21, gadd45 expression and p53 binding activity, as well as a significant decline in DNA repair capability and accelerated telomere shortening. Our results suggest that multiple pathways might be involved in oxidative SIPS, including genes related to DNA-damage-and-repair and telomere shortening, and that SIPS shares the same mechanisms with replicative senescence in vivo. Our findings indicate that several aging theories can be merged together by a common mechanism of oxidative damage, and that the level of oxidative DNA damage-and-repair capacity may be exploited as reliable markers of cell senescence. PMID- 15833274 TI - Subcellular recruitment by TSG118 and TSPYL implicates a role for zinc finger protein 106 in a novel developmental pathway. AB - To gain insight into the function of zinc finger protein 106 (ZFP106), we analyzed its subcellular targeting and identified its interacting proteins. Although ZFP106 was detected predominantly in the fibrillar component of the nucleolus and co-localized with the nucleolar transcriptional machinery, its overexpression did not affect transcription of pre-ribosomal RNA genes. The nucleolar association of ZFP106 did neither require ongoing ribosomal RNA synthesis nor nucleolar chromatin indicating that a protein-protein interaction confines ZFP106 to the nucleolus. Deletion analysis revealed that the C-terminal WD40 repeat region functions in nucleolar targeting. This domain interacts with the product of testis-specific gene 118 (TSG118), which also co-localizes with ZFP106 in the nucleolus. Rapid downregulation of TSG118 expression during in vitro terminal differentiation coincides with a loss of nucleolar ZFP106. By its structural features and expression, TSG118 mimics nucleostemin, a nucleolar protein linked to the proliferation potential of stem cells. A two-hybrid screen with the N-terminal region of ZFP106 as bait led to the isolation of testis specific Y-encoded-like protein (TSPYL), a member of the nucleosome assembly protein family. A frame-shift mutation in TSPYL has recently been found to cause a sudden infant death syndrome with testis dysgenesis. Specific recruitment of ZFP106 via amino acids 412-781 into TSPYL-positive nucleoplasmic bodies requires a TSPYL domain absent in the mutant protein of patients with testis dysgenesis. These results identify ZFP106 as a potential player in a novel pathway involved in testis development. PMID- 15833275 TI - Investigation of methaemoglobin reduction by extracellular NADH in mammalian erythrocytes. AB - The effect of extracellular NADH on the rate of reduction of nitrite-induced methaemoglobin in erythrocytes from man, cattle, dog, horse, grey kangaroo, pig and sheep was investigated. Extracellular NADH was found to enhance the rate of methaemoglobin reduction in man, dog, pig and kangaroo erythrocytes, but had essentially no effect on the rate of methaemoglobin reduction in erythrocytes from cattle, horse and sheep. In erythrocytes of those animals affected by extracellular NADH the rate of reduction of metHb in the presence of NADH was the same or greater than that observed in the presence of nutrients such as glucose and inosine. The combination of nutrient and NADH produced a more profound increase in the rate of methaemoglobin reduction. The rate of methaemoglobin reduction in all cases was significantly less than that observed with methylene blue, the standard treatment of methaemoglobinaemia. Extracellular NADH was found to indirectly increase the intracellular NADH concentration through displacement of the pseudo-equilibrium of the intracellular LDH reaction and relied upon the presence of sufficient LDH activity released into the extracellular medium through haemolysis. The lack of response of cattle, horse and sheep RBCs to extracellular NADH was found to derive mainly from their low extracellular LDH activity, but also correlated with their lower NADH-methaemoglobin reductase activity compared to the other species. PMID- 15833276 TI - Chemical modification studies on alkaline phosphatase from pearl oyster (Pinctada fucata): a substrate reaction course analysis and involvement of essential arginine and lysine residues at the active site. AB - Alkaline phosphatases (ALP, EC 3.1.3.1) are ubiquitous enzymes found in most species. ALP from a pearl oyster, Pinctada fucata (PALP), is presumably involved in nacreous biomineralization processes. Here, chemical modification was used to investigate the involvement of basic residues in the catalytic activity of PALP. The Tsou's plot analysis indicated that the inactivation of PALP by 2,4,6 trinitrobenzenesulfonic acid (TNBS) and phenylglyoxal (PG) is dependent upon modification of one essential lysine and one essential arginine residue, respectively. Substrate reaction course analysis showed that the TNBS and PG inactivation of PALP followed pseudo-first-order kinetics and the second-order inactivation constants for the enzyme with or without substrate binding were determined. It was found that binding substrate slowed the PG inactivation whereas had little effect on TNBS inactivation. Protection experiments showed that substrates and competitive inhibitors provided significant protection against PG inactivation, and the modified enzyme lost its ability to bind the specific affinity column. However, the TNBS-induced inactivation could not be prevented in presence of substrates or competitive inhibitors, and the modified enzyme retained the ability to bind the affinity column. In a conclusion, an arginine residue involved in substrate binding and a lysine residue involved in catalysis were present at the active site of PALP. This study will facilitate to illustrate the role ALP plays in pearl formation and the mechanism involved. PMID- 15833277 TI - NADPH oxidase mediates interleukin-6 expression in cerulein-stimulated pancreatic acinar cells. AB - NADPH oxidase produces a large amount of reactive oxygen species (ROS) mainly in phagocytic cells. ROS are involved in NF-kappaB activation, cytokine expression and thus, pathogenesis of pancreatitis. However, the source of ROS in pancreatic acinar cells has not been clarified. Cerulein rapidly induces acute and edematous form of pancreatitis. We investigated whether pancreatic acinar cells contain NADPH oxidase, and whether NADPH oxidase mediates interleukin-6 (IL-6) in pancreatic acinar AR42J cells stimulated with cerulein. Expression of NADPH oxidase subunits and NADPH oxidase activity were determined in the cells by immunofluorescence staining and lucigenin luminescence, respectively. Oxidant sensitive nuclear transcription factor NF-kappaB activation was monitored by electrophoretic mobility shift assay. IL-6 expression was determined by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbant assay. NADPH oxidase inhibitor diphenylene iodonium (DPI), antioxidant rebamipide, and antisense oligonucleotides (AS ODNs) for NADPH oxidase subunits p22phox and p47phox were used to determine the involvement of NADPH oxidase in NF-kappaB activation and IL-6 expression in AR42J cells. As a result, pancreatic acinar AR42J cells constitutively express NADPH oxidase subunits p67phox and p47phox in the cytosol and Nox1 and p22phox in the membrane. Cerulein-stimulated NADPH oxidase activity and induced NF-kappaB activation and IL-6 expression in AR42J cells. Treatment of DPI or rebamipide and transfection of AS ODNs for NADPH oxidase subunits suppressed cerulein-induced NF-kappaB activation and IL-6 expression compared to S ODNs. In conclusion, NADPH oxidase may mediate the expression of inflammatory cytokines by stimulating NF-kappaB activation in pancreatic acinar cells during the course of pancreatitis. PMID- 15833278 TI - Smooth muscle alpha-actinin interaction with smitin. AB - Actin-myosin II filament-based contractile structures in striated muscle, smooth muscle, and nonmuscle cells also contain the actin filament-crosslinking protein alpha-actinin. In striated muscle sarcomeres, interactions between the myosin binding protein titin and alpha-actinin in the Z-line provide an important structural linkage. We previously discovered a titin-like protein, smitin, associated with the contractile apparatus of smooth muscle cells. Purified native smooth muscle alpha-actinin binds with nanomolar affinity to smitin in smitin myosin coassemblies in vitro. Smooth muscle alpha-actinin also interacts with striated muscle titin. In contrast to striated muscle alpha-actinin interaction with titin and smitin, which is significantly enhanced by PIP2, smooth muscle alpha-actinin interacts with smitin and titin equally well in the presence and absence of PIP2. Using expressed regions of smooth muscle alpha-actinin, we have demonstrated smitin-binding sites in the smooth muscle alpha-actinin R2-R3 spectrin-like repeat rod domain and a C-terminal domain formed by cryptic EF-hand structures. These smitin-binding sites are highly homologous to the titin-binding sites of striated muscle alpha-actinin. Our results suggest that direct interaction between alpha-actinin and titin or titin-like proteins is a common feature of actin-myosin II contractile structures in striated muscle and smooth muscle cells and that the molecular bases for alpha-actinin interaction with these proteins are similar, although regulation of these interactions may differ according to tissue. PMID- 15833279 TI - The modulation of apoptosis by cyclic AMP involves Akt and epidermal growth factor receptor. AB - Adenosine 3',5'-cyclic monophosphate (cAMP) and transforming growth factor-beta are important regulators of many biological processes. In this study we investigated the effect and its potential mechanism of cAMP on transforming growth factor-beta1- and serum deprivation-induced apoptosis in Mv1Lu cells. Transforming growth factor-beta1 treatment or serum deprivation induces apoptotic response in Mv1Lu cells. Forskolin, a cAMP-elevating agent, or 8-Bromo-cAMP (8-B cAMP), a cell permeable cAMP analogue, inhibited the cell proliferation and markedly enhanced apoptosis induced by transforming growth factor-beta1, but completely suppressed serum deprivation-induced apoptosis. Furthermore, forskolin decreased the Akt phosphorylation, and the inhibition of phosphatidylinositol-3 kinase by LY294002 sensitized Mv1Lu cells to transforming growth factor-beta1 induced apoptosis. In addition, forskolin treatment induced tyrosine phosphorylation of epidermal growth factor receptor. Inhibition of epidermal growth factor receptor by specific inhibitor PD153035 blocked the cAMP-mediated suppression of serum deprivation-induced apoptosis. The results indicate that cAMP exerts its opposite effects in transforming growth factor-beta1- and serum deprivation-induced apoptosis via a mechanism involving the modulation of signaling components of phosphatidylinositol-3-kinase/Akt and epidermal growth factor receptor in Mv1Lu cells. PMID- 15833280 TI - Human gastric signet ring carcinoma (KATO-III) cell apoptosis induced by Vitex agnus-castus fruit extract through intracellular oxidative stress. AB - We have previously reported that an ethanol extract of the dried ripe fruit of Vitex agnus-castus (Vitex) displays cytotoxic activity against certain kinds of human cancer cell line resulting in the induction of apoptosis. In this paper, we investigate the molecular mechanism of apoptosis induced by Vitex using a human gastric signet ring carcinoma cell line, KATO-III. DNA fragmentation was observed in Vitex-treated KATO-III cells in a time- and dose-dependent manner. DNA fragmentation was accompanied by the following phenomena: elevation in the level of hemeoxygenase-1 protein and thioredoxin reductase mRNA; repression of Mn superoxide dismutase and catalase mRNAs; release of cytochrome c from mitochondria into the cytosol; activation of caspases-8, -9 and -3; decrease in the level of Bcl-2, Bcl-XL and Bid protein; increase in the level of Bad protein. The intracellular oxidized state, measured using 2',7'-dichlorofluorescin diacetate, increased after Vitex treatment. While the amount of intracellular GSH decreased significantly after treatment with Vitex, the level of GSSG was unaffected. Furthermore, no significant perturbation in the amount of proteins/mRNAs related to glutathione metabolism could be detected. These apoptotic alterations induced by exposure to Vitex were blocked by the presence of an anti-oxidative reagent, N-acetyl-l-cysteine, or the addition of exogenous GSH. Our results demonstrate that intracellular oxidative stress and mitochondrial membrane damage is responsible for Vitex-induced apoptosis, which may be mediated by a diminution of reduced type glutathione within the cell. PMID- 15833281 TI - The cellular distribution and stage-specific expression of two dynein light chains from the human blood fluke Schistosoma japonicum. AB - Schistosomes are pathogenic helminth parasites of human portal veins. Their body wall is a highly active syncytial tegument involved in an array of host interactions. The cytoskeletal organization and dynamics of this syncytium are poorly understood, but predominant motor components are the LC8 class of cytoplasmic dynein light chains (DLC). Four LC8 members occur in schistosomes, two of which are expressed in the tegument. Here, we describe the cytoplasmic distribution, stage-specific expression and cellular location of two diverse LC8 molecules of Schistosoma japonicum. SjDLC1 was detected in surface-membrane specific extracts of adult worms and was shown by quantitative immuno-electron microscopy to predominate along heptalaminate membranes of the worm surface. SjDLC3 also occurs in the tegument, but was shown to be present in basal layers of the tegument and did not preferentially co-localize with particular membrane components. SjDLC3 was also detected in the gastrodermis. SjDLC1 is expressed only in mammalian-parasitic stages, whereas SjDLC3 is expressed throughout the life-cycle. The data suggest that SjDLC1 is preferentially located to the host interactive distal parasite membrane, and plays a role in surface membrane dynamics, while SjDLC3 is a ubiquitous motor component of schistosome epithelia of all stages. PMID- 15833282 TI - Carbohydrate-specifically polyethylene glycol-modified ricin A-chain with improved therapeutic potential. AB - Ricin A-chain, which exhibits excellent cytotoxicity to tumor cells, has been widely used as an immunotoxin source. However, it has the fatal shortcoming of poor pharmacokinetics due to the tremendous liver uptake via carbohydrate mediated recognition. Modification of proteins with polyethylene glycol, PEGylation, has the advantages of shielding the specific sites and prolonging the biological half-life. In this study, the carbohydrate-specific PEGylation of ricin A-chain was considered to be a novel approach to overcome this limitation. The carbohydrate group of ricin A-chain was oxidized by sodium m-periodate and further specifically conjugated with hydrazide-derivatized PEG. For a comparative study, the PEGylated ricin A-chain at amino groups was prepared using the hydroxysuccinimide ester-derivatized PEG. The carbohydrate-specifically PEGylated ricin A-chain showed a markedly lower liver uptake and systemic clearance compared with the amine-directly PEGylated ricin A-chain as well as the unmodified ricin A-chain. Furthermore, carbohydrate-specifically PEGylated ricin A-chain showed a significantly higher in vitro ribosome-inactivating activity than the amine-directly PEGylated ricin A-chain. These findings clearly demonstrate that the carbohydrate-specificity as well as PEGylation plays an important role in improving the in vivo pharmacokinetic properties and in vitro bioactivity. Therefore, these results suggest that the therapeutic use of immunotoxins constructed using this carbohydrate-specifically PEGylated ricin A chain has potential as a cancer therapy. PMID- 15833283 TI - Peroxisome proliferator-activated receptor alpha and its response element are required but not sufficient for transcriptional activation of the mouse heart type fatty acid binding protein gene. AB - Expression of heart-type fatty acid binding protein is restricted mainly to the skeletal and cardiac muscles and further regulated by peroxisome proliferator activated receptor alpha. The molecular basis for the muscle-restricted peroxisome proliferator-activated receptor alpha action on the fatty acid binding gene was analyzed using normal and the receptor-null mice and the cultured cells. Two possible peroxisome proliferator-response elements were found in the promoter region of the mouse gene. A gel shift assay showed that both elements were functional. However, neither the tandem repeats of the elements nor the cloned promoter sequence could be activated by peroxisome proliferator-activated receptor alpha and its ligand in the reporter gene assay using cultured cells. The cloned promoter responded to the ligand only in the muscle when the reporter gene was introduced into the mouse muscle. Using a chimeric receptor with the activation domain of herpes virus VP16 protein and the tandem repeats of the elements with or without mutation, the upstream element was finally demonstrated to be potentially involved in the receptor-dependent transcriptional activation. These results suggest that the peroxisome proliferator-response element of the mouse gene is atypical and there is a muscle-specific mechanism to enhance the weak binding of the receptor to the response element to ensure the muscle specific action of peroxisome proliferator-activated receptor alpha on the heart type fatty acid binding protein gene promoter. PMID- 15833284 TI - Characterization by liquid chromatography combined with mass spectrometry of monoclonal anti-IGF-1 receptor antibodies produced in CHO and NS0 cells. AB - 7H2HM is a new humanized recombinant monoclonal antibody (MAb) directed against insulin-like growth factor-1 receptor and produced in CHO cells. Homogeneity of intact antibody, reduced light and heavy chains, Fab and Fc fragments were investigated by analytical methods based on mass (SDS-PAGE, SEC), charge (IEF, C IEX) and hydrophobicity differences (RP-HPLC, HIC) and compared side-by-side with A2CHM, produced in NS0 cells. Primary structures and disulfide bridge pairing were analyzed by microsequencing (Edman degradation), mass spectrometry (MALDI TOF, ES-TOF) and peptide mapping after enzymatic digestion (Trypsin, endoprotease Lys-C, papain). The light chains demonstrated the expected sequences. The heavy chains yielded post-translational modifications previously reported for other recombinant humanized or human IgG1 such as N-terminal pyroglutamic acid, C terminal lysine clipping and N-glycosylation for asparagine 297. More surprisingly, two-thirds of the 7H2HM heavy chains were shown to contain an additional 24-amino-acid sequence, corresponding to the translation of an intron located between the variable and the constant domains. Taken together these data suggest that 7H2HM is a mixture of three families of antibodies corresponding (i) to the expected structure (17%; 14,9297 Da; 1330 amino acids), (ii) a variant with a translated intron in one heavy chains (33%; 15,2878 Da; 1354 amino acids) and (iii) a variant with translated introns in two heavy chains (50%; 15,4459 Da; 1378 amino acids), respectively. RP-HPLC is not a commonly used chromatographic method to assess purity of monoclonal antibodies but unlike to SEC and SDS-PAGE, was able to show and to quantify the family of structures present in 7H2HM, which were also identified by peptide mapping, mass spectrometry and microsequencing. PMID- 15833285 TI - Analysis of protein fragmentation inhibition by an MMP-inhibitor in an in vivo model of heart failure using automated chromatography. AB - Proteomic strategies have continued to demonstrate value in studying disease by exploiting new technologies that can develop significant numbers of measurements from single samples. However, using complex samples such as tissues or blood has continued to be problematic due to the presence of major interfering substances. In this study, a process is described that uses denaturing peptide extraction from whole tissue and automated chromatography in order to allow subsequent analysis of more than 1000 tissue-derived peptides per sample. The process was employed to identify cardiac proteins that were spared degradation by administration of a heart-protecting matrix metalloproteinase (MMP) inhibitor (compound SC-621) following experimental myocardial infarction (MI). HPLC peptide fingerprints were developed from rat heart left ventricles and the resultant integrated peak data was compared across experimental animals. Surprisingly, although protein fragmentation was generally increased in MI hearts, the effect of the MMP inhibitor was only observed on a few species. The results from this study demonstrated that whole-tissue sample enrichment and peptide analysis using HPLC could be linked in order to study the effects of new compounds on a disease state. The system is flexible and amenable to improvements such as incorporating detection by mass spectrometry. PMID- 15833286 TI - Simultaneous determination of 16 purine derivatives in urinary calculi by gradient reversed-phase high-performance liquid chromatography with UV detection. AB - A reversed-phase high-performance liquid chromatography (HPLC) method with ultraviolet detection has been developed for the analysis of purines in urinary calculi. The method using gradient of methanol concentration and pH was able to separate 16 compounds: uric acid, 2,8-dihydroxyadenine, xanthine, hypoxanthine, allopurinol and oxypurinol as well as 10 methyl derivatives of uric acid or xanthine (1-, 3-, 7- and 9-methyluric acid, 1,3-, 1,7- and 3,7-dimethyluric acid, 1-, 3- and 7-methylxanthine). Limits of detection for individual compounds ranged from 0.006 to 0.035 mg purine/g of the stone weight and precision (CV%) was 0.5 2.4%. The method enabled us to detect in human uric acid stones admixtures of nine other purine derivatives: natural metabolites (hypoxanthine, xanthine, 2,8 dihydroxyadenine) and methylated purines (1-, 3- and 7-methyluric acid, 1,3 dimethyluric acid, 3- and 7-methylxanthine) originating from the metabolism of methylxanthines (caffeine, theophylline and theobromine). The method allows simultaneous quantitation of all known purine constituents of urinary stones, including methylated purines, and may be used as a reference one for diagnosing disorders of purine metabolism and research on the pathogenesis of urolithiasis. PMID- 15833287 TI - Determination of the cyanide metabolite 2-aminothiazoline-4-carboxylic acid in urine and plasma by gas chromatography-mass spectrometry. AB - The cyanide metabolite 2-aminothiazoline-4-carboxylic acid (ATCA) is a promising biomarker for cyanide exposure because of its stability and the limitations of direct determination of cyanide and more abundant cyanide metabolites. A simple, sensitive, and specific method based on derivatization and subsequent gas chromatography-mass spectrometry (GC-MS) analysis was developed for the identification and quantification of ATCA in synthetic urine and swine plasma. The urine and plasma samples were spiked with an internal standard (ATCA-d(2)), diluted, and acidified. The resulting solution was subjected to solid phase extraction on a mixed-mode cation exchange column. After elution and evaporation of the solvent, a silylating agent was used to derivatize the ATCA. Quantification of the derivatized ATCA was accomplished on a gas chromatograph with a mass selective detector. The current method produced a coefficient of variation of less than 6% (intra- and interassay) for two sets of quality control (QC) standards and a detection limit of 25 ng/ml. The applicability of the method was evaluated by determination of elevated levels of ATCA in human urine of smokers in relation to non-smokers for both males and females. PMID- 15833288 TI - Development of a high-performance liquid chromatography-tandem mass spectrometry method for the determination of flurogestone acetate in ovine plasma. AB - Flurogestone (FGA) is a synthetic progesterone, with a progestational action higher than that of progesterone itself. It is intended for vaginal use in large animals to induce oestrus synchronization. A quantitative method for the analysis of flurogestone acetate (FGA) in ovine plasma by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been developed. After the incorporation of megestrol acetate (MGA) as internal standard (IS) and followed by a liquid-liquid extraction from plasma, FGA and MGA were chromatographed using a reverse-phase HPLC column and detected by tandem mass spectrometry with a TurboIonSpray source. Multiple reaction-monitoring (MRM) mode was used for the quantitative determination of FGA in ovine plasma. The precursor ions [M+H](+) at m/z 407.2 and 385.1 for FGA and MGA, respectively, produced product ions at m/z 267.1/285.1 for FGA and m/z 267.1/224.0 for MGA. The validated concentration range was 0.2-5.0 ng/ml based on 500 microl plasma aliquots. The lower limit of quantitation was 0.2 ng/ml. Fully validated selectivity, accuracy, precision and reproducibility criteria for routine use in pharmacokinetic studies were demonstrated. PMID- 15833289 TI - Determination and occurrence of polybrominated diphenyl ethers in maternal adipose tissue from inhabitants of Singapore. AB - Polybrominated diphenyl ethers (PBDEs), as a specific group of brominated flame retardants (BFR), are used in a variety of consumer products including electronics and household furnishings. In recent years, a marked increase in the levels of PBDEs in human biological tissues and fluids, especially breast milk, has been reported in several countries. However, few data are available from countries in the Asia-pacific region, including Singapore. This study presents a validated method procedure and the first available data of the concentrations of PBDE congeners: PBDE-47 (2,2,4,4-Tetrabromodiphenyl ether), PBDE-99 (2,2',4,4',5 Pentabromodiphenyl ether), PBDE-100 (2,2',4,4',6-Pentabromodiphenyl ether), PBDE 153 (2,2',4,4',5,5'-Hexabromodiphenyl ether), PBDE-154 (2,2',4,4',5,6' Hexabromodiphenyl ether) in maternal adipose tissue collected from inhabitants of Singapore. Microwave-assisted extraction (MAE) of PBDEs spiked adipose tissues coupled with GC-MS analysis achieved comparable recoveries to a conventional Soxhlet Extraction (SE) procedure of between 70 and 130%. MAE also yielded comparable precision data (variance less than 13%) relative to the SE procedure. Spiked Carbon-13 PBDE congeners were also used as surrogates for MAE quality assurance and confirmed the efficiency of the procedure. PBDE congeners were detected in all of 16 maternal adipose tissues collected in Singapore, where levels were comparable to available data from Belgium. PMID- 15833290 TI - Intracellular measurements of anti-HIV drugs indinavir, amprenavir, saquinavir, ritonavir, nelfinavir, lopinavir, atazanavir, efavirenz and nevirapine in peripheral blood mononuclear cells by liquid chromatography coupled to tandem mass spectrometry. AB - A sensitive and accurate liquid chromatography-tandem mass spectrometric (LC MS/MS) method for the intracellular determination of nine antiretroviral drugs in human peripheral blood mononuclear cells (PBMCs) is proposed. PBMCs are isolated by density gradient centrifugation using Vacutainer CPT tubes and cell count is performed with a Coulter instrument. Single-step extraction of drugs from PBMCs pellets is performed with MeOH 50% (with clozapine added as internal standard, I.S.) and the supernatant is injected onto a 2.1 mm x 30 mm SymmetryShield 3.5 microm-RP18 column equipped with a 2.1 x 10 mm guard column. Chromatographic separations are performed using a gradient program with a mixture of 2 mM ammonium acetate containing 0.1% formic acid and acetonitrile with 0.1% formic acid. Analytes quantification is performed by electro-spray ionisation-triple quadrupole mass spectrometry using the selected reaction monitoring (SRM) detection mode. The positive mode is used for the HIV protease inhibitors (PIs) indinavir, amprenavir, saquinavir, ritonavir, nelfinavir, lopinavir, atazanavir and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine, and the negative mode is applied for efavirenz. The calibration curves are prepared using blank PBMCs spiked with antiretroviral drugs at concentrations ranging from 0.5 to 100 ng/ml of cell extracts and fitted to a quadratic regression model weighted by 1/(concentration)(2). The lower limit of quantification is less than 0.5 ng/ml. The mean extraction recovery for all PIs/NNRTIs is always above 88%. The method is precise, with mean inter-day CV% within 0.6-10.2%, and accurate (range of inter-day deviation from nominal values -7.2 to +8.3%). This analytical method can be conveniently used in clinical research for the assessment of intracellular levels of all PIs/NNRTIs commercially available at present using a simple one-step cell extraction of PBMCs followed by liquid chromatography coupled with tandem triple quadripole mass detection. PMID- 15833291 TI - Sensitive liquid chromatography-tandem mass spectrometry method for the determination of cefixime in human plasma: application to a pharmacokinetic study. AB - A sensitive and selective liquid chromatographic-tandem mass spectrometric (LC-MS MS) method was developed to determine cefixime ((6R,7R)-7-[(Z)-2-(2-amino-4 thiazolyl)-2-(carboxymethoxyimino)acetamido]-8-oxo-3-vinyl-5-thia-1-azabicyclo [4,2,0]-oct-2-ene-2-carboxylic acid) in human plasma. After a simple protein precipitation using acetonitrile, the post-treatment samples were analyzed on a C(8) column interfaced with a triple quadrupole tandem mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase consisted of acetonitrile-water-formic acid (40:60:0.5, v/v/v). The analyte and internal standard cefetamet were both detected by use of selected reaction monitoring mode. The method was linear in the concentration range of 0.05-8.0 microg/ml. The lower limit of quantification was 0.05 microg/ml. The intra- and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 12.7%. The accuracy determined at three concentrations (0.05, 0.80 and 7.2 microg/ml for cefixime) was within +/ 2.0% in terms of relative error. Each plasma sample was chromatographed within 3.5 min. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of cefixime capsule in 24 healthy volunteers. PMID- 15833292 TI - Characterization of lipophilicity and antiproliferative activity of E-2 arylmethylene-1-tetralones and their heteroanalogues. AB - A molecular library based on E-2-arylmethylene-1-tetralone has been designed and synthesized. A reversed phase high performance liquid chromatographic (RP-HPLC) method has been developed and applied to separate them and to characterize their lipophilicity. The chromatographic method applied here was suitable to separate the structural (ortho and para) isomers of compounds and was sensitive enough to differentiate their lipophilicities. The measured (k') and computer calculated (CLOGP) lipophilicity values has been compared. Good linear correlation has been found in the case of these structurally related molecules. In vitro biological assay has been performed with Methylene blue dying to investigate the antiproliferative potency of the compounds synthesized in this work. The measured (k') and calculated (CLOGP) lipophilicities of the compounds were compared with the antiproliferative activities and an optimum value of lipophilicity has been found for these compounds. PMID- 15833293 TI - Fully automated liquid-liquid extraction for the determination of a novel insulin sensitizer in human plasma by heated nebulizer and turbo ionspray liquid chromatography-tandem mass spectrometry. AB - I, 2-{[5,7-dipropyl-3-(trifluoromethyl)-1,2-benzisoxazol-6-yl]oxy}-2-methyl propionic acid is an alpha peroxisome proliferator-activated receptor (PPAR) agonist with some gamma activity being investigated for potential use in the treatment of Type II diabetes mellitus and dyslipidemia. Two automated liquid liquid extraction methods were developed and validated for the determination of I in human plasma. Concentrations of I were determined over a wide range of clinical doses. For Method A, plasma was acidified and extracted with ethyl acetate using a fully automated procedure. Analysis was performed by LC-MS/MS with a turbo ionspray source in negative ion mode. For Method B, a larger volume of plasma was extracted and a heated nebulizer source was used on the mass spectrometer. Method A was linear from 0.05 to 50 ng/mL and Method B from 0.2 to 1000 ng/mL. Validation procedures showed that both methods were robust, specific and reproducible. PMID- 15833294 TI - Adsorption of bilirubin with polylysine carrying chitosan-coated nylon affinity membranes. AB - Microporous polyamide membranes were activated by bisoxirane and subsequently bound with chitosan (CS) to amplify reactive groups. Then polylysine (PLL) as ligand was immobilized onto the CS-coated nylon membranes. The contents of CS and PLL of PLL-attached membranes were 93.2 and 90.4 mg/g nylon membrane, respectively. Such PLL-attached membranes were used to adsorb bilirubin from the bilirubin-phosphate solution and bilirubin-albumin solution. The adsorption mechanism of bilirubin and the effects of temperature, initial concentration of bilirubin, albumin concentration and ionic strength on adsorption were investigated by batch experiments. The results showed that the adsorption capacity increased with increasing the temperature while decreased with increasing the NaCl concentration and albumin concentration, and the adsorption isotherm fitted the Freundlich model well. The result of dynamic experiment showed PLL-attached membranes can well remove the bilirubin from the bilirubin albumin solution. PMID- 15833295 TI - Novel and simple high-performance liquid chromatographic method for determination of 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity. AB - We present here a high-performance liquid chromatographic method for the evaluation of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. The automated method was applied to fungal and mouse liver extracts and validated by the addition of mevastatin to the reaction mixture and by several intra- and inter-day assays. This method offers important advantages over those previously reported because no radiolabeled substrates or expensive techniques such as mass spectrometry are required, and the time of analysis is relatively short. Moreover, the method can be successfully applied to different biological samples; hence, it should be very useful in evaluating potential inhibitors of the HMG-CoA enzyme and investigating cholesterol metabolism, cell growth and differentiation processes. PMID- 15833296 TI - Development of simultaneous gas chromatography-mass spectrometric and liquid chromatography-electrospray ionization mass spectrometric determination method for the new designer drugs, N-benzylpiperazine (BZP), 1-(3 trifluoromethylphenyl)piperazine (TFMPP) and their main metabolites in urine. AB - To prove the intake of recently controlled designer drugs, N-benzylpiperazine (BZP) and 1-(3-trifluoromethylphenyl)piperazine (TFMPP), a simple, sensitive and reliable method which allows us to simultaneously detect BZP, TFMPP and their major metabolite in human urine has been established by coupling gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). GC-MS accompanied by trifluoroacetyl (TFA) derivatization and LC-MS analyses were performed after the enzymatic hydrolysis and the solid phase extraction with OASIS HLB, and BZP, TFMPP and their major metabolites, 4'-hydroxy-BZP (p-OH-BZP), 3'-hydroxy-BZP (m-OH-BZP) and 4'-hydroxy-TFMPP (p-OH-TFMPP), have found to be satisfactorily separated on a semi-micro SCX column with acetonitrile-40 mM ammonium acetate buffer (pH 4) (75:25, v/v) as the eluent. The detection limits produced by GC-MS were estimated to be from 50 ng/ml to 1 microg/ml in the scan mode, and from 200 to 500 ng/ml in the selected ion monitoring (SIM) mode. Upon applying the LC-ESI-MS technique, the linear calibration curves were obtained by using the SIM mode for all analytes in the concentration range from 10 ng/ml to 10 microg/ml. The detection limits ranged from 5 to 40 ng/ml in the scan mode, and from 0.2 to 1 ng/ml in the SIM mode. These results indicate the high reliability and sensitivity of the present procedure, and this procedure will be applicable for proof of intake of BZP and TFMPP in forensic toxicology. PMID- 15833297 TI - Improved enantioselective analysis of polyunsaturated hydroxy fatty acids in psoriatic skin scales using high-performance liquid chromatography. AB - Enantioselective analysis is used as a valuable tool for determining the biological origin of chiral derivatives of arachidonic, 11,14-eicosadienoic and linoleic acid in psoriatic skin scales and for clarifying their role in pathogenesis. This paper reports on a simple and rapid enantioselective determination (without any derivatization) of the fatty acid derivatives 13(R,S) hydroxyoctadecadienoic acid [13(R,S)-HODE], 9(R,S)-hydroxyoctadecadienoic acid [9(R,S)-HODE] and 12(R,S)-hydroxyeicosatetraenoic acid [12(R,S)-HETE], using high performance liquid chromatography (HPLC) with Chiralpak AD as the chiral selector and electrospray ionisation mass spectrometry (ESI-MS). The enantiomeric distribution of 12-HETE, 9-HODE and 13-HODE in psoriatic skin scales of untreated patients (untreated during the last 4 weeks before sampling) was evaluated in comparison to psoriatic skin scales of patients underlying systemic treatment. The enantiomeric distribution of 12-HETE and 9-HODE showed no remarkable differences, whilst samples of patients under systemic treatment exhibited a lower predominance of 13(S)-HODE than samples of untreated patients. Furthermore, the effect of UVB phototherapy on the enantiomeric distribution of 12-HETE, 9 HODE and 13-HODE was studied and a semiquantitation of these compounds in psoriatic skin scales performed. The detected amounts of 9-HODE in samples of untreated patients were remarkably lower than those in samples of patients underlying systemic treatment. In the case of UVB phototherapy, no influence on the enantiomeric distribution could be observed. PMID- 15833298 TI - Quantitative determination of perfluorooctanoic acid in serum and plasma by liquid chromatography tandem mass spectrometry. AB - A selective and sensitive method for analysis of perfluorooctanoic acid (PFOA) in human serum and plasma, utilizing liquid chromatography tandem mass spectrometry (LC-MS/MS), has been developed and thoroughly validated to satisfy strict FDA guidelines for bioanalytical methods. A simple, automated sample preparation procedure, involving extraction of the target analyte with acetonitrile on protein precipitation media in a 96-well plate format was developed, allowing efficient handling of large numbers of samples. The proposed method uses the calibration standards prepared in a surrogate matrix (rabbit serum or plasma) and (13)C-labeled PFOA as the internal standard to account for matrix effects, instrument drift, and extraction efficiency. Human serum and plasma could not be used for matrix matching of calibration standards as endogenous levels of PFOA observed in the control human serum and plasma significantly exceeded the targeted lower limit of quantitation (LLOQ) of the method. Precision and accuracy of the method were demonstrated by analysis of rabbit serum and plasma control samples fortified at 0.5, 5, and 40 ng/mL PFOA and human serum and plasma fortified at 1.0, 5.0, 40 ng/mL PFOA. The LLOQ of 0.5 ng/mL PFOA was experimentally demonstrated for rabbit and human serum and plasma. Within-day precision and accuracy, short-term stability, freeze-thaw stability, equivalence of response between PFOA and APFO (the ammonium salt of PFOA), and dilution of concentrated samples were also investigated. The results of the validation experiments comply with the precision and accuracy limits defined by the FDA guidance document: "Guidance for Industry, Bioanalytical Method Validation", May 2001. PMID- 15833299 TI - Evaluation of evaporative light-scattering detection for metabolite quantification without authentic analytical standards or radiolabel. AB - Development of a sensitive and specific technique for the quantitation of drug metabolites without the use of synthetic analytical standards or radiolabel would represent a major advance in preliminary route of metabolism screening in drug discovery. In this study, the ability of evaporative light-scattering detection (ELSD) to quantify metabolites of 7-ethoxycoumarin (EC) was evaluated. Because ELSD operates as a mass detector, the complex nature of in vitro-derived samples from hepatocyte incubations resulted in an inability to detect the analytes of interest in this matrix using ELSD. Additionally, the gradient nature of the analysis required to temporally separate ethoxycoumarin from its metabolites and matrix components interfered with the ELSD response. Furthermore, using less complex contrived mixtures, ELSD demonstrated insufficient sensitivity (limit of detection of 1000-10,000 ng/mL) and an inconsistent inter-analyte response. Together, the limitations outlined in these experiments demonstrate that ELSD is at present an inadequate technique for generating semi-quantitative data on metabolites in drug discovery. PMID- 15833300 TI - Reply to comment on: "Assessment of nitric oxide biosynthesis and peroxynitrite formation within the central nervous system by measuring L-citrulline in the cerebrospinal fluid?". PMID- 15833301 TI - Action experience alters 3-month-old infants' perception of others' actions. AB - An intervention facilitated 3-month-old infants' apprehension of objects either prior to (reach first), or after (watch first) viewing another person grasp similar objects in a visual habituation procedure. Action experience facilitated action perception: reach-first infants focused on the relation between the actor and her goal, but watch-first infants did not. Infants' sensitivity to the actor's goal was correlated with their engagement in object-directed contact with the toys. These findings indicate that infants can rapidly form goal-based action representations and suggest a developmental link between infants' goal directed actions and their ability to detect goals in the actions of others. PMID- 15833302 TI - Audio-visual speech perception is special. AB - In face-to-face conversation speech is perceived by ear and eye. We studied the prerequisites of audio-visual speech perception by using perceptually ambiguous sine wave replicas of natural speech as auditory stimuli. When the subjects were not aware that the auditory stimuli were speech, they showed only negligible integration of auditory and visual stimuli. When the same subjects learned to perceive the same auditory stimuli as speech, they integrated the auditory and visual stimuli in a similar manner as natural speech. These results demonstrate the existence of a multisensory speech-specific mode of perception. PMID- 15833303 TI - Word meaning and the control of eye fixation: semantic competitor effects and the visual world paradigm. AB - When participants are presented simultaneously with spoken language and a visual display depicting objects to which that language refers, participants spontaneously fixate the visual referents of the words being heard [Cooper, R. M. (1974). The control of eye fixation by the meaning of spoken language: A new methodology for the real-time investigation of speech perception, memory, and language processing. Cognitive Psychology, 6(1), 84-107; ]. We demonstrate here that such spontaneous fixation can be driven by partial semantic overlap between a word and a visual object. Participants heard the word 'piano' when (a) a piano was depicted amongst unrelated distractors; (b) a trumpet was depicted amongst those same distractors; and (c), both the piano and trumpet were depicted. The probability of fixating the piano and the trumpet in the first two conditions rose as the word 'piano' unfolded. In the final condition, only fixations to the piano rose, although the trumpet was fixated more than the distractors. We conclude that eye movements are driven by the degree of match, along various dimensions that go beyond simple visual form, between a word and the mental representations of objects in the concurrent visual field. PMID- 15833304 TI - Horizontal and vertical Simon effect: different underlying mechanisms? AB - Reaction times are usually faster when stimulus and response occur at the same location than when they do not, even if stimulus location is irrelevant to the task (Simon effect). This effect was found with both horizontal and vertical stimulus-response arrangements. The same mechanisms have been proposed to be involved in either case. Here, we compared a horizontal and a vertical Simon task by means of a RT time-course analysis of the Simon effect. Also, we analysed the lateralised readiness potential (LRP), an index of covert response-preparation processes. In the horizontal task, the Simon effect decays over time and pre activation occurs above the motor cortex ipsilateral to the stimulus. In contrast, the Simon effect does not decay over time and no early incorrect LRP deflection is observed in the vertical task. These findings suggest that typical activation accounts can fit only the horizontal Simon effect, while a translation explanation is more suitable for the vertical Simon effect. PMID- 15833305 TI - Starting at the end: the importance of goals in spatial language. AB - We explored the linguistic encoding of Paths in children between the ages of three and seven, in children with Williams syndrome, and in normal adults, focusing specifically on Source and Goal Paths. The results showed an asymmetry, with Goal Paths regularly and systematically encoded, but Source Paths often omitted. This pattern occurred among all groups and across a broad range of domains including Manner of Motion, Change of Possession, Change of State, and Attachment/Detachment events. It also occurred whether participants spontaneously described events or were asked to use a specific verb that biased them towards a Goal or Source Path (e.g. 'give' vs. 'get'). The results are discussed in terms of non-linguistic foundations of spatial language and the linguistic mapping biases that arise when we describe what we see. PMID- 15833306 TI - The categorical structure of semantic memory for famous people: a new approach using release from proactive interference. AB - Memory for familiar people is essential to understand their identity and guide social interaction. Nevertheless, we know surprisingly little about the structure of such memory. Previous research has assumed that semantic memory for people has a categorical structure, but recently it was proposed that memory for people consists only of associations and lacks any categorical structure. Four experiments are reported that use a novel approach by adapting the 'release from proactive interference' (RPI) methodology for use with lists of famous names. Proactive interference occurs when items presented on successive trials are drawn from the same category. Recall can improve following a change to a different category. Sets of names were selected relating to aspects previously demonstrated, on the basis of reaction time data, to form a category (occupation) and a property (nationality) of celebrities (Johnston & Bruce, 1990). RPI was observed for a change at both levels of representation but was only present without explicitly cueing the change of set when the stimuli differed at the category level. At the property level, RPI was only evident when change of set was explicitly cued. RPI was absent at the set change in a novel, ad hoc distinction suggesting that the effect reflected the underlying memory structure. PMID- 15833307 TI - The relationship between object files and conscious perception. AB - Object files (OFs) are hypothesized mid-level representations which mediate our conscious perception of persisting objects-e.g. telling us 'which went where'. Despite the appeal of the OF framework, not previous research has directly explored whether OFs do indeed correspond to conscious percepts. Here we present at least one case wherein conscious percepts of 'which went where' in dynamic ambiguous displays diverge from the analogous correspondence computed by the OF system. Observers viewed a 'bouncing/streaming' display in which two identical objects moved such that they could have either bounced off or streamed past each other. We measured two dependent variables: (1) an explicit report of perceived bouncing or streaming; and (2) an implicit 'object-specific preview benefit' (OSPB), wherein a 'preview' of information on a specific object speeds the recognition of that information at a later point when it appears again on the same object (compared to when it reappears on a different object), beyond display wide priming. When the displays were manipulated such that observers had a strong bias to perceive streaming (on over 95% of the trials), there was nevertheless a strong OSPB in the opposite direction-such that the object files appeared to have 'bounced' even though the percept 'streamed'. Given that OSPBs have been taken as a hallmark of the operation of object files, the five experiments reported here suggest that in at least some specialized (and perhaps ecologically invalid) cases, conscious percepts of 'which went where' in dynamic ambiguous displays can diverge from the mapping computed by the object-file system. PMID- 15833308 TI - A comment on noise enhanced bursting in pancreatic beta-cells. PMID- 15833309 TI - Modeling reciprocation and cooperation in primates: evidence for a punishing strategy. AB - Experiments in which animals strategically interact with one another or search over some controlled domain are becoming common. While these experiments often promise to illuminate sophisticated animal behavior, the analyses brought to bear on these data are often quite coarse. For example, many papers simply tally the number of observations consistent with a behavioral theory. This analysis is simple, but ignores a potentially rich source of information by failing to take into account patterns and systematic variation among observations inconsistent with the theory. Using a new data set generated by cotton-top tamarin monkeys playing a repeated food-exchange game, we apply a maximum-likelihood estimation technique (more commonly used to study human economic behavior) which utilizes much more of the information in these data, and which uncovers unexpectedly sophisticated cooperative behavior from our subjects. Tamarin cooperation remains stable as long as both actors consistently cooperate, but requires at least two consecutive unexpected acts of cooperation to restart cooperation after it has collapsed, a strategy that resembles two-tits for a tat. We conclude by enumerating the benefits of a maximum-likelihood approach in experimental settings such as ours, and suggest other areas in which these techniques may be fruitful. PMID- 15833310 TI - A mathematical model of cellular apoptosis and senescence through the dynamics of telomere loss. AB - The shortening of telomeric repeats as a cell replicates has long been implicated as a determinant of cell viability. However, recent studies have indicated that it is not telomere length, but rather whether telomeres have bound a telomere related protein, which in mammals is TTAGGG repeat binding factor-2 (TRF2), that determines whether a cell undergoes apoptosis (programmed cell death), enters senescence (a quiescent, non-replicative state), or continues to proliferate. When bound to a telomere, TRF2 allows a cell to recognize the telomere as the point where a chromosome ends rather than a break in DNA. When telomeres are not bound by TRF2, the cell can either immediately trigger senescence or apoptosis via the DNA damage response pathway, or indirectly trigger it by attempting to repair the chromosome, which results in chromosomal end joining. We model the ability of telomeres to bind TRF2 as a function of telomere length and apply the resulting binding probability to a model of cellular replication that assumes a homogeneous cell population. The model fits data from cultured human fibroblasts and human embryonic kidney cells for two free parameters well. We extract values for the percent of telomere loss at which cell proliferation ceases. We show, in agreement with previous experiments, that overexpression of TRF2 allows a cell to delay the senescence setpoint. We explore the effect of oxidative stress, which increases the rate of telomere loss, on cell viability and show that cells in the presence of oxidative stress have reduced lifespans. We also show that the addition of telomerase, an enzyme that maintains telomere length, is sufficient to result in cell immortality. We conclude that the increasing inability of TRF2 to bind telomeres as they shorten is a quantitatively reasonable model for a cause of either cellular apoptosis or senescence. PMID- 15833311 TI - The strategy of ecosystem development: specific dissipation as an indicator of ecosystem maturity. AB - The ratio of entropy generation rate to entropy embodied in structures relatively to the surroundings can be considered as an indicator of the ability of a self organizing dissipative system to maintain itself far from equilibrium by pumping out entropy. The higher the ratio (which may be called the specific entropy production or the specific dissipation of a system), the lower the capacity of a system to convert the incoming low-entropy energy into internal organization. It appears that the ratio attains special significance for interpreting the evolution of biological systems, as the maximum expression of self-organizing systems, from the sub-cellular to the ecosystem scale. This paper proposes specific dissipation, written as the ratio of biological entropy production to exergy stored in the living biomass, as a thermodynamic orientor as well as an indicator of the development state of ecological systems. After having presented a method for estimating the specific dissipation in lakes, the adequacy of the proposed indicator is discussed and also tested by comparing its response to those of some classical ecological attributes (successional sequences of species, biodiversity, individual body size, structural organization and generation time of organisms) throughout the seasonal progression of the plankton community in Lake Trasimeno (Umbria, Italy). The results support the hypothesis that the minimization of specific dissipation is a primary criterion of evolution of ecological systems and also sustain the use of specific dissipation as an indicator of ecological maturity. PMID- 15833312 TI - Neural cross-correlation and signal decorrelation: insights into coding of auditory space. AB - The auditory systems of humans and many other species use the difference in the time of arrival of acoustic signals at the two ears to compute the lateral position of sound sources. This computation is assumed to initially occur in an assembly of neurons organized along a frequency-by-delay surface. Mathematically, the computations are equivalent to a two-dimensional cross-correlation of the input signals at the two ears, with the position of the peak activity along this surface designating the position of the source in space. In this study, partially correlated signals to the two ears are used to probe the mechanisms for encoding spatial cues in stationary or dynamic (moving) signals. It is demonstrated that a cross-correlation model of the auditory periphery coupled with statistical decision theory can predict the patterns of performance by human subjects for both stationary and motion stimuli as a function of stimulus decorrelation. Implications of these findings for the existence of a unique cortical motion system are discussed. PMID- 15833313 TI - Intestinal propulsion of a solid non-deformable bolus. AB - A mathematical model of a segment of the gut with an enclosed pellet is constructed. The gut is represented as a thin deformable soft biological shell with the pellet modeled as a non-deformable solid. Mechanical properties of the gut wall were represented as longitudinal and circular smooth muscle layers embedded in stroma that satisfies the general type of nonlinear orthotropy. Deformations of the wall are finite. Bolus propulsion is numerically simulated by generation and propagation of an electromechanical wave along the syncytia. Pharmacological manipulation is applied to model 5-HT type 3 antagonist (Lotronex, GlaxoSmithKline) and 5-HT type 4 agonist (Zelnorm, Novartis, AB) drugs on the dynamics of bolus progression. The results lead to new quantitative insights into the complex spatio-temporal patterns of gastrointestinal transit. It is demonstrated that the reciprocal relationship in contraction of the longitudinal and circular smooth muscle syncytia is necessary to provide the "mixing" type of movements during the preparatory phase of propulsion. Strong simultaneous contractions of the both smooth muscle layers are required to expel the "mixed" pellet from the segment. The model is implemented as an interactive software system, Gut Discovery(www.aincompany.com), and accurately predicts the effects of drugs on gut motility. PMID- 15833314 TI - Fingerprint formation. AB - Fingerprints (epidermal ridges) have been used as a means of identifications for more than 2000 years. They have also been extensively studied scientifically by anthropologists and biologists. However, despite all the empirical and experimental knowledge, no widely accepted explanation for the development of epidermal ridges on fingers, palms and soles has yet emerged. In this article we argue that fingerprint patterns are created as the result of a buckling instability in the basal cell layer of the fetal epidermis. Analysis of the well known von Karman equations informs us that the buckling direction is perpendicular to the direction of greatest stress in the basal layer. We propose that this stress is induced by resistance of furrows and creases to the differential growth of the basal layer and regression of the volar pads during the time of ridge formation. These ideas have been tested by computer experiments. The results are in close harmony with observations. Specifically, they are consistent with the well-known observation that the pattern type is related to the geometry of the fingertip surface when fingerprint patterns are formed. PMID- 15833315 TI - Maintenance of handedness polymorphism in humans: a frequency-dependent selection model. AB - Frequency-dependent selection is an important process in the maintenance of genetic variation in fitness. In humans, it has been proposed that the polymorphism of handedness is maintained by negative frequency-dependent selection, through a strategic advantage of left-handers in fighting interactions. Using simple mathematical models, we explore: (1) whether it is possible to predict the range of left-handedness frequencies observed in human populations by the frequency and the violence of fighting interactions; (2) the consequences of the sex differences in the probability of transmission of hand preference to offspring. We show that a wide range of values of the frequency of left-handers can be obtained with realistic changes of the parameters values. Our models reinforce the idea that negative frequency-dependence may have played a role in maintaining left-handedness in human populations, and provide further support for the importance of fighting interactions in the evolution of hand preference. Moreover, they suggest an explanation for the occurrence of left handedness among women in this context, namely an indirect selective advantage through their male offspring. PMID- 15833316 TI - Keystone species and vulnerable species in ecological communities: strong or weak interactors? AB - The loss of a species from an ecological community can trigger a cascade of secondary extinctions. The probability of secondary extinction to take place and the number of secondary extinctions are likely to depend on the characteristics of the species that is lost--the strength of its interactions with other species- as well as on the distribution of interaction strengths in the whole community. Analysing the effects of species loss in model communities we found that removal of the following species categories triggered, on average, the largest number of secondary extinctions: (a) rare species interacting strongly with many consumers, (b) abundant basal species interacting weakly with their consumers and (c) abundant intermediate species interacting strongly with many resources. We also found that the keystone status of a species with given characteristics was context dependent, that is, dependent on the structure of the community where it was embedded. Species vulnerable to secondary extinctions were mainly species interacting weakly with their resources and species interacting strongly with their consumers. PMID- 15833317 TI - Influence of fracture gap size on the pattern of long bone healing: a computational study. AB - Following fractures, bones restore their original structural integrity through a complex process in which several cellular events are involved. Among other factors, this process is highly influenced by the mechanical environment of the fracture site. In this study, we present a mathematical model to simulate the effect of mechanical stimuli on most of the cellular processes that occur during fracture healing, namely proliferation, migration and differentiation. On the basis of these three processes, the model then simulates the evolution of geometry, distributions of cell types and elastic properties inside a healing fracture. The three processes were implemented in a Finite Element code as a combination of three coupled analysis stages: a biphasic, a diffusion and a thermoelastic step. We tested the mechano-biological regulatory model thus created by simulating the healing patterns of fractures with different gap sizes and different mechanical stimuli. The callus geometry, tissue differentiation patterns and fracture stiffness predicted by the model were similar to experimental observations for every analysed situation. PMID- 15833318 TI - The effect of residual Ca2+ on the stochastic gating of Ca2+-regulated Ca2+ channel models. AB - Single-channel models of intracellular Ca(2+) channels such as the inositol 1,4,5 trisphosphate receptor and ryanodine receptor often assume that Ca(2+)-dependent transitions are mediated by a constant background [Ca(2+)] as opposed to a dynamic [Ca(2+)] representing the formation and collapse of a localized Ca(2+) domain. This assumption neglects the fact that Ca(2+) released by open intracellular Ca(2+) channels may influence subsequent gating through the processes of Ca(2+)-activation or -inactivation. We study the effect of such "residual Ca(2+)" from previous channel opening on the stochastic gating of minimal and realistic single-channel models coupled to a restricted cytoplasmic compartment. Using Monte Carlo simulation as well as analytical and numerical solution of a system of advection-reaction equations for the probability density of the domain [Ca(2+)] conditioned on the state of the channel, we determine how the steady-state open probability (p(open)) of single-channel models of Ca(2+) regulated Ca(2+) channels depends on the time constant for Ca(2+) domain formation and collapse. As expected, p(open) for a minimal model including Ca(2+) activation increases as the domain time constant becomes large compared to the open and closed dwell times of the channel, that is, on average the channel is activated by residual Ca(2+) from previous openings. Interestingly, p(open) for a channel model that is inactivated by Ca(2+) also increases as a function of the domain time constant when the maximum domain [Ca(2+)] is fixed, because slow formation of the Ca(2+) domain attenuates Ca(2+)-mediated inactivation. Conversely, when the source amplitude of the channel is fixed, increasing the domain time constant leads to elevated domain [Ca(2+)] and decreased open probability. Consistent with these observations, a realistic De Young-Keizer-like IP(3)R model responds to residual Ca(2+) with a steady-state open probability that is a monotonic function of the domain time constant, though minimal models that include both Ca(2+)-activation and -inactivation show more complex behavior. We show how the probability density approach described here can be generalized for arbitrarily complex channel models and for any value of the domain time constant. In addition, we present a comparatively simple numerical procedure for estimating p(open) for models of Ca(2+)-regulated Ca(2+) channels in the limit of a very fast or very slow Ca(2+) domain. When the ordinary differential equation for the [Ca(2+)] in a restricted cytoplasmic compartment is replaced by a partial differential equation for the buffered diffusion of intracellular Ca(2+) in a homogeneous isotropic cytosol, we find the dependence of p(open) on the buffer time constant is qualitatively similar to the above-mentioned results. PMID- 15833319 TI - Decavanadate effects in biological systems. AB - Vanadium biological studies often disregarded the formation of decameric vanadate species known to interact, in vitro, with high-affinity with many proteins such as myosin and sarcoplasmic reticulum calcium pump and also to inhibit these biochemical systems involved in energy transduction. Moreover, very few in vivo animal studies involving vanadium consider the contribution of decavanadate to vanadium biological effects. Recently, it has been shown that an acute exposure to decavanadate but not to other vanadate oligomers induced oxidative stress and a different fate in vanadium intracellular accumulation. Several markers of oxidative stress analyzed on hepatic and cardiac tissue were monitored after in vivo effect of an acute exposure (12, 24 h and 7 days), to a sub-lethal concentration (5 mM; 1 mg/kg) of two vanadium solutions ("metavanadate" and "decavanadate"). It was observed that "decavanadate" promote different effects than other vanadate oligomers in catalase activity, glutathione content, lipid peroxidation, mitochondrial superoxide anion production and vanadium accumulation, whereas both solutions seem to equally depress reactive oxygen species (ROS) production as well as total intracellular reducing power. Vanadium is accumulated in mitochondria in particular when "decavanadate" is administered. These recent findings, that are now summarized, point out the decameric vanadate species contributions to in vivo and in vitro effects induced by vanadium in biological systems. PMID- 15833320 TI - Controllable preparation of Nano-MgO and investigation of its bactericidal properties. AB - Samples of nano-MgO with varying particle sizes were prepared by four different methods using Mg(NO3)2.6H2O, Na2CO3, urea and ammonia as raw materials and characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), low temperature N2 adsorption-desorption measurements and FT-IR spectroscopy. Bactericidal experiments with Bacillus subtilis var. niger and Staphylococcus aureus were carried out using as-synthesized nano-MgO samples and the bactericidal mechanism was also investigated. The results showed that the bactericidal efficacy of nano-MgO increases with decreasing particle size. The bactericidal efficacy of the samples was compared with that of TiO2, a common photoactive bactericidal material. The nano-MgO has better bactericidal activity, both when used directly and as an additive in an interior wall paint. Furthermore, nano-MgO is active even in the absence of irradiation. PMID- 15833321 TI - [Ru(phen)2DPPZ]2+ is in contact with DNA bases when it forms a luminescent complex with single-stranded oligonucleotides. AB - The luminescence intensity of the Delta- and Lambda-enantiomer of [Ru(phen)2DPPZ]2+ ([Ru(phenanthroline)2 dipyrido[3,2-a:2',3'-c]phenazine]2+) complex enhanced upon binding to double stranded DNA, which has been known as "light switch effect". The enhancement of the luminescence required the intercalation of the large ligand between DNA base pairs. In this study, we report the enhancement in the luminescence intensity when the metal complexes bind to single stranded oligonucleotides, indicating that the "light switch effect" does not require intercalation of the large DPPZ ligand. Oligonucleotides may provide a hydrophobic cavity for the [Ru(phen)2DPPZ]2+ complex to prevent the quenching by the water molecule. In the cavity, the metal complex is in contact with DNA bases as is evidenced by the observation that the excited energy of the DNA bases transfer to the bound metal complex. However, the contact of the metal complex with DNA bases is different from the stacking of DPPZ in the intercalation pocket. In addition to the normal two luminescence lifetimes, a short lifetime in the range of 1-2 ns was found for both the delta- and lambda enantiomer of [Ru(phen)2DPPZ]2+ when complexed with single stranded oligonucleotides, which may be assigned to the metal complex that is outside of the cavity, interacting with phosphate groups of DNA. PMID- 15833322 TI - Synthesis and characterization of a tetracycline-platinum (II) complex active against resistant bacteria. AB - A tetracycline-platinum(II) complex, [PtCl2(C22H24N2O8)], was synthesized and characterized by elemental analysis, conductivity and thermogravimetric analyses, and infrared spectroscopy. The interaction of tetracycline (Tc) with platinum(II) ions was also studied in aqueous solution by 1H NMR and circular dichroism spectroscopies. Tetracycline forms a 1:1 complex with platinum via the oxygen of the hydroxyl group at the A ring and that of the amide group. The complex is as efficient as tetracycline in inhibiting the growth of two Escherichia coli sensitive bacterial strains and six times more potent against E. coli HB101/pBR322, a bacterial strain resistant to tetracycline. This finding is very important because the use of tetracycline to treat bacterial infections has declined due to the emergence of resistant organisms. PMID- 15833323 TI - A spectroscopic investigation of the self-association and DNA binding properties of a series of ternary ruthenium(II) complexes. AB - Six ruthenium(II) complexes of the general form cis-alpha-[Ru(N4-tetradentate)(N2 bidentate)]Cl2 have been synthesized from the two related tetradentate ligands 1,6-di(2'-pyridyl)-2,5-dimethyl-2,5-diazahexane (picenMe2) and 1,6-di(2'-pyridyl) 2,5-dibenzyl-2,5-diazahexane (picenBz2) and the bidentate ligands 2,2'-bipyridine (bipy), 1,10-phenanthroline (phen) and dipyrido[3,2-f:2'3'-h]quinoxaline (dpq). Synthetic intermediate species of the general form cis-alpha-[Ru(II)(N4 tetradentate)(DMSO)Cl][PF6] were isolated. The N4-tetradentate ligand picenMe2 formed only the cis-alpha stereoisomer, while picenBz2 formed both the cis-alpha and cis-beta stereoisomers. These latter stereoisomers were resolved by fractional crystallisation. Dimer self-association constants, K(D), were estimated from the concentration dependence of the 1H NMR shifts for some of these complexes in aqueous solutions at 25 degrees C. The values of K(D) ranged from 0.6 to 7.9 M(-1) and a relationship was observed between the aromatic surface area of the bidentate component and the degree to which self-association occurred, whereby a greater level of self-association correlates with a larger surface area for the bidentate ligand. Some of these complexes demonstrate an ability to bind to DNA that is consistent with intercalation of the bidentate molecular component between the base pairs of the DNA molecule. Using calf-thymus DNA, the equilibrium binding constants, K(B), were determined for some of the complexes using intrinsic methods and these ranged from 3.32 to 5.11 M(-1), the intercalating abilities of the different bidentate ligands being in the order dp q > phen > bipy. This relationship between aromatic surface area of the bidentate ligand and the degree of DNA binding activity is the same as that observed in the self-association study. PMID- 15833324 TI - A non-radioactive method for measuring Cu uptake in HepG2 cells. AB - At present, all data on Cu uptake and metabolism have been derived from radioactive uptake experiments. These experiments are limited by the availability of the radioactive isotopes 64Cu or 67Cu, and their short half-life (12.5 and 62 h, respectively). In this paper, we investigate an alternative method to study the uptake of Cu with natural isotopes in HepG2 cells, a liver cell line used extensively to study Cu metabolism. In nature, Cu occurs as two stable isotopes, 63Cu and 65Cu (63Cu/65Cu = 2.23). This ratio can be measured accurately using inductively coupled plasma mass spectrometry (ICP-MS). In initial experiments, we attempted to measure the time course of Cu uptake using 65Cu. The change in the 63Cu/65Cu ratio, however, was too small to allow measurement of Cu uptake by the cells. To overcome this difficulty, the natural 63Cu/65Cu ratio in HepG2 cells was altered using long-term incubation with 63Cu. This had a significant effect on Cu concentration in HepG2 cells, changing it from 81.9 +/- 9.46 pmol microg DNA(-1) (week 1) to 155 +/- 8.63 pmol microg DNA(-1) (week 2) and stabilising at 171 +/- 4.82 pmol microg DNA(-1) (week 3). After three weeks of culture with 2 microM 63Cu the 63Cu/65Cu changed from 2.18 +/- 0.05 to 15.3 +/- 1.01. Cu uptake was then investigated as before using 65Cu. Uptake was linear over 60 min, temperature dependent and consistent with previous kinetics data. These observations suggest that stable isotope ICP-MS provides an alternative technique for the study of Cu uptake by HepG2 cells. PMID- 15833325 TI - Antibacterial activity of highly negative charged polyoxotungstates, K27[KAs4W40O140] and K18[KSb9W21O86], and Keggin-structural polyoxotungstates against Helicobacter pylori. AB - The antibacterial activity of polyoxometalates (PMs) against Helicobacter pylori was investigated based on determinations of minimum inhibitory concentration (MIC) and fractional inhibitory concentration (FIC), time-killing of the bacteria, bacterial morphology and PM-uptake into the bacteria cell. The result of MIC values revealed that, of 13 PMs used in this study, highly negative charged polyoxotungstates, such as K27[KAs4W40O140] and K18[KSb9W21O86], and Keggin-structural polyoxotungstates exhibited a potent antibacterial activity with the MIC values of less than 256 microg/ml. The former was the most active, and superior to metronidazole (MTZ) against MTZ-susceptible and resistant strains and also to clarithromycin (CLR) against CLR-resistant strains. In contrast, most of polyoxomolybdates showed little antibacterial activity with the MIC values of more than 256 microg/ml. The result of FIC index values indicated that the antibacterial polyoxotungstates had partially synergistic effect in combination with MTZ and CLR but indifferent effect in combination with amoxicillin (AMX). From the results of the time-killing and scanning electron microscope images, K27[KAs4W40O140] and K18[KSb9W21O86] proved the concentration-dependent bactericidal activity with the morphological change from bacillary form to coccoid form, while Keggin-structural K5[SiV(V)W11O40] showed the bacteriostatic activity with small change of morphology to coccoid form. The fluorescent X-ray analysis demonstrated that these polyoxotungstates were taken into the bacteria cell. It is pointed out that the Keggin-structure and/or high negativity polyoxotungstates are an important factor for the antibacterial activity against H. pylori. PMID- 15833326 TI - Functional models for catechol dioxygenases: iron(III) complexes of cis-facially coordinating linear 3N ligands. AB - A series of 1:1 iron(III) complexes of simple and sterically hindered tridentate 3N donor ligands have been synthesized and studied as functional models for catechol dioxygenases. All of them are of the type [FeLCl3], where L is bis(pyrid 2-yl-methyl)amine (L1), N,N-bis(benzimidazol-2-ylmethyl)amine (L2), N-methyl-N' (pyrid-2-ylmethyl)ethylenediamine (L3), N,N-dimethyl-N'-(pyrid-2-ylmethyl) ethylenediamine (L4) and N-phenyl-N'-(pyrid-2-ylmethyl)ethylenediamine (L5). They have been characterised by spectral and electrochemical methods. The X-ray crystal structure of the complex [Fe(L4)Cl3] has been successfully determined. The complex crystallizes in the triclinic space group P1 with a = 7.250(6), b = 8.284(3), c = 12.409(4) angstroms, alpha = 80.84(3) degrees, beta = 86.76(6) degrees, gamma = 72.09(7) degrees and Z = 2. It possesses a distorted octahedral geometry in which the L4 ligand is cis-facially coordinated to iron(III) and the chloride ions occupy the remaining coordination sites. The systematic variation in the ligand donor atom type significantly influences the Lewis acidity of the iron(III) center and hence the binding interaction of the complexes with simple and substituted catechols. The spectroscopic and electrochemical properties of the catecholate complexes generated in situ have been investigated. All the complexes catalyze mainly the oxidative intradiol cleavage of 3,5-di-tert butylcatechol (H2DBC) in the presence of dioxygen, which is unexpected of the cis facial coordination of the ligands. The rate of intradiol catechol cleavage reaction depends upon the Lewis acidity of iron(III) center and steric demand and hydrogen-bonding functionalities of the ligands. Interestingly, the electron-sink property of N-phenyl substituent in [Fe(L5)Cl3] complex leads to enhancement in rate of cleavage. All these observations provide support to the substrate activation mechanism proposed for intradiol-cleaving enzymes. PMID- 15833327 TI - Adsorption of recombinant human bone morphogenetic protein rhBMP-2m onto hydroxyapatite. AB - Recombinant human mature bone morphogenetic protein 2 (rhBMP-2m) has been expressed to study its adsorption onto precipitated hydroxyapatite (HA). The influence on the adsorption process of different parameters such as pH and concentrations of calcium, phosphate or NaCl has been investigated. Although the adsorption proceeds rapidly at the initial stages, the maximum adsorbed amount is reached after four hours. The process is notably influenced by adding calcium or phosphate to the system but, while calcium ions increase the adsorption of rhBMP 2m onto HA, phosphate ions inhibit it. The influence of pH and NaCl concentration are notable but less important than those of calcium and phosphate. The adsorption data fit well to a Langmuir adsorption isotherm. The values of the isotherm parameters have been calculated and discussed, and an adsorption mechanism has been proposed. PMID- 15833328 TI - Electrochemical reduction of cytochrome P450 as an approach to the construction of biosensors and bioreactors. AB - In the present review an attempt was made to present an up-to-date amount of the data on electrochemical reduction of the hemoprotein cytochrome P450. The concept and potentialities of enzyme electrodes--transducers--as the main element for construction of electrochemical biosensors were discussed. Different types of electrodes for bioelectrochemistry were analysed. New nanotechnological approaches to cytochrome P450 immobilisation were reported. It was shown that nanobiotechnology in electrochemistry has potential application in manufacturing biosensors and bioreactors for clinical medicine and pharmacology. PMID- 15833329 TI - Pharmacodynamics and pharmacokinetics of the insulin-mimetic agent vanadyl acetylacetonate in non-diabetic and diabetic rats. AB - The objectives of this study were to evaluate the pharmacodynamics and pharmacokinetics of vanadyl acetylacetonate (VAC) in rats. Pharmacodynamic study was carried out using non-diabetic and diabetic rats by subcutaneous (s.c.) and intragastric (i.g.) administrations at single dose or multiple doses. Pharmacokinetic study was performed using non-diabetic rats. Results showed that VAC resulted in a significant decrease of plasma glucose levels in diabetic rats in all dosing levels, and nearly restored hyperglycemic values to normal values after s.c. injection at a single dose of 2, 4, and 8 mg vanadium (V)/kg, or after i.g. administration at multiple doses of 3 and 6 mg V/kg once daily for seven consecutive days, respectively. The VAC could be rapidly absorbed and T(max) values ranged from 0.9 +/- 0.3 h for s.c. injection to 3.0 +/- 0.9 h for i.g. administration. The average absolute bioavailabilities for i.g. administrations at a single dose of 3, 6, and 10 mg V/kg were 34.7%, 28.1%, and 22.8%, respectively. After i.g. administration at a single dose of 10 mg V/kg, the average elimination half-lives obtained from non-diabetic rats were very long ranging from 144.7 +/- 8.7 h in plasma to 657.3 +/- 34.8 h in femur tissue. In conclusion, VAC widely distributed in various tissues and accumulated more in the femur tissue. The time to reach maximal vanadium level after s.c. injection or i.g. administration was not coincident with the time to reach maximal hypoglycemic effect. The accumulated vanadium in bone, kidney or other tissues may gradually release and exert a longer action. In present dosing levels and administration routes, VAC was effective for lowering plasma glucose levels in diabetic rats and could reverse the higher triglyceride and cholesterol levels to the normal ranges. VAC did not influence the insulin levels in plasma and not cause obvious toxic signs like diarrhea. PMID- 15833330 TI - Synthesis, crystal structure and nuclease activity of a Schiff base copper(II) complex. AB - A new Schiff base copper(II) complex, Cu(o-VANAHE)(2) (o-VANAHE = 2-(o vanillinamino)-1-hydroxyethane), has been synthesized and characterized. Single crystal X-ray diffraction results suggest that this complex structure belongs to triclinic crystal system, space group P1 with the following crystallographic parameters: a = 8.819(4) angstroms, b = 10.794(5) angstroms, c = 11.350(5) angstroms, alpha = 70.262(6) degrees, beta = 70.816(6) degrees, gamma = 78.360(6) degrees, V = 955.4(7) angstroms3, Z = 2, D(c) = 1.571 Mg x m(-3), and the final R1 = 0.0393, wR2 = 0.0994 for the observed reflections 2620(I > 2sigma(I)). The molecular geometry is almost coplanar. Viscosity, fluorescence spectroscopy and cyclic voltammetry have been conducted to assess their interaction between this complex and DNA. Results showed that the copper(II) complex can increase DNA's relative viscosity and quench the fluorescence intensity of EB bound to DNA. The adding of DNA to the solution of Cu(o-VANAHE)2 causes a slight decrease in the voltammetric current, as well as a slight shift in the E(1/2) to less negative potential. The interaction between the complex and DNA has also been investigated by submarine gel electrophoresis, interestingly, we found that the copper(II) complex can cleave circular plasmid pBR322 DNA to nicked and linear forms. PMID- 15833331 TI - Sodium orthovanadate exerts influence on liver Golgi complexes from control and streptozotocin-diabetic rats. AB - The paper presents the effect of one-week 3mM sodium orthovanadate (Na3VO4) oral treatment of control and streptozotocin[STZ]-diabetic rats. The body weight decreased as compared with untreated control (C group) in both vanadate treated groups (C + V and D + V) and in diabetic untreated rats (D group)--in all cases p < 0.01. A similar tendency was demonstrated by the weight of the livers, which was statistically significant lower than in the controls (p < 0.01). The fluid and food intake were lower in comparison with control vanadium treated groups, in D + V as compared with D it was limited, however, not achieved control level. A high mortality rate, approx. 67%, after the administration of streptozotocin and vanadate (D + V group) was noted; such result had never been previously found within all study groups of rats. But the surviving rats show very good decreased (60%) free blood sugar levels, however euglycaemia was not achieved. The activity of galactosyltransferase, the Golgi complex marker enzyme in group D, was statistically lower than the controls (p < 0.001). Treatment of STZ-diabetic rats with orthovanadate did not increase the enzyme activity toward control level, in both diabetic groups (treated and untreated with Na3VO4) similar dispersion of individual results was found. Morphological study demonstrated, for the first time, no larger cellular lesion in C + V group. The Golgi complex was well developed; showed several cisterns at the trans side, which were grossly distended and contained electron-lucid floccular material. In D + V group typical, cylindrical forms of Golgi complexes predominated. These structures consisted of 3-4 almost practically non-distended cisterns. Also in this case, large, electron-dense vesicles were noted in the vicinity. In this group, small in size, myelin-like structures were also found. These structures might indicate a relatively small, but nevertheless clear damage of the internal membrane system. The external cistern of the cylindrical forms of Golgi complexes, which corresponded the trans side, was often markedly distended and formed a vacuole like structure filled with electron lucent material; the structure itself sometimes looked empty. Multi-vesicular structures were observed also in this case, but they were seen much more rarely. PMID- 15833332 TI - Interactions between metal ions and carbohydrates. The coordination behavior of neutral erythritol to neodymium ion. AB - A single crystal of a coordinated complex of neutral erythritol (C4H10O4,E) with a neodymium ion, NdE(II), was synthesized and studied using FT-IR and X-ray diffraction analysis. In NdE(II) (NdCl3.2.5C4H10O4.C2H5OH) the Nd3+ coordinates with one chloride ion and eight OH groups from three erythritol molecules. There are two neodymium centers linked by one erythritol molecule with same coordination structure in the molecule. Two erythritol molecules provide 1,3,4 hydroxyl groups to coordinate with a neodymium ion; another erythritol molecule coordinates to two Nd ions via its 1,2-hydroxyl groups and 3,4-hydroxyl groups, respectively. The OH groups of erythritol act as ligand to coordinate to neodymium ions, and OH groups of erythritol form hydrogen bond networks that link chain and layer together to build three-dimensional structures. The ratio of metal to ligand is 1:2.5. The structure of NdE(II) is more complicated than the previously reported NdE(I), which is NdCl3.C4H10O4.6H2O; in NdE(I), Nd3+ is coordinated to four hydroxyl groups from two erythritol molecules, four water molecules and one chloride ion. The results indicate the complexity of metal sugar interaction. PMID- 15833333 TI - Synthesis, characterization and binding with DNA of four planaramineplatinum(II) complexes of the forms: trans-PtL2Cl2 and [PtL3Cl]Cl, where L = 3 hydroxypyridine, 4-hydroxypyridine and imidazo(1,2-alpha)pyridine. AB - Four new trans-planaramineplatinum(II) complexes, three of the form: trans PtCl2L2, code named CH1, CH2 and CH4 where L = 3-hydroxypyridine, 4 hydroxypyridine and imidazo[1,2-alpha]pyridine, respectively, and one of the form: PtClL3, code named CH3 where L = 3-hydroxypyridine, have been prepared and characterized by elemental analyses and IR, Raman, mass and 1H NMR spectral studies. The interactions of the compounds with salmon sperm and pBR322 plasmid DNAs have been investigated and their activity against human ovarian cancer cell lines: A2780, A2780cisR and A2780ZD0473R have also been determined. The compounds are believed to form mainly monofunctional N7(G) and bifunctional intrastrand N7(G)N7(G) adducts with DNA, causing a local distortion of DNA as a result of which gel mobility of the DNA changes. The compound containing three planaramine ligands per molecule (CH3) is found to be less reactive than the compounds containing two planaramine ligands per molecule (CH1, CH2 and CH4), which in turn are less reactive than compounds containing one of the same planaramine ligands per molecule. The decrease in reactivity is reflected in lower molar conductivity values (indicating lower degree of dissociation), less pronounced changes caused to DNA conformation (indicating decreased level of platinum-DNA binding) and lower activity. The decreased reactivity of the compounds is due to a greater steric crowding produced by the bulky planaramine ligands. Changes in DNA conformation are also found to be a function of the actual nature of the planaramine ligand. The results illustrate structure-activity relationship. PMID- 15833334 TI - Mixed-ligand Ru(II) complexes with 2,2'-bipyridine and aryldiazo-beta-diketonato auxillary ligands: synthesis, physico-chemical study and antitumour properties. AB - The complexes of Ru(II)-2,2'-bipyridyl with substituted diazopentane-2,4-diones (L1H-L5H) were synthesized and characterized by elemental analyses, conductance, FAB (fast atom bombardment) mass and spectral (IR, UV/Vis (UV/visible), NMR) studies. Molecular geometry optimization of the complexes was also made. None of the complexes luminesce. However, facilitated oxidation of Ru(II) to Ru(III) was evidenced from their lower reduction potential data. The ligands and their complexes were tested for their antitumour activity against a variety of tumour cell lines. Though activity is found to vary with the type of tumour cell lines used, yet complex 5 with naphtyldiazopentane-2,4-dione as co-ligand was found to be a potential compound as it showed in general significant activity against all cell lines studied. PMID- 15833335 TI - Synthesis, crystal structure, cytotoxic activity and DNA-binding properties of the copper (II) and zinc (II) complexes with 1-[3-(2-pyridyl)pyrazol-1 ylmethyl]naphthalene. AB - A new ligand L, 1-[3-(2-pyridyl)pyrazol-1-ylmethyl]naphthalene, and its two metal complexes, [Cu(L)3](ClO4)2 (1) and [Zn(L)3](ClO4)2(H2O)2 (2), have been synthesized and characterized. The crystal structure of complex 1 was determined by single crystal X-ray diffraction, which crystallized in monoclinic, space group P2(1)/n with unit cell parameters, a = 12.710(4) angstroms, b = 12.135(3) angstroms, c = 33.450(9) angstroms, beta = 93.281(5) degrees and Z = 4. The Cu atom was six-coordinated to N(1), N(2), N(4), N(5), N(7) and N(8) from three L ligands and formed a slightly distorted octahedral geometry. Complexes 1 and 2, and ligand L were subjected to biological tests in vitro using three different cancer cell lines (HL-60, BGC-823 and MDA-MB-435). Complex 1 showed significant cytotoxic activity against three cancer cell lines. The interactions of complexes 1 and 2, and ligand L with calf thymus DNA were then investigated by thermal denaturation, viscosity measurements and spectrophotometric methods. The experimental results indicated that complexes 1 and 2 bound to DNA by intercalative mode via the ligand L. The intrinsic binding constants of complexes 1 and 2, and ligand L with DNA were 1.8 x 10(4), 5.4 x 10(3) and 2.76 x 10(3) M( 1), respectively. PMID- 15833336 TI - Racemic D,L-[Co(phen)2dpq](3+)-DNA interactions: investigation into the basis for minor-groove binding and recognition. AB - A study on the minor-groove recognition of B-DNA d(GTCGAC)2 by racemic D,L [Co(phen)2dpq](3+), where phen and dpq stand for 1,10-phenanthroline and dipyrido [3,2-d:2,3-f]quinoxaline, respectively, was carried out with a one-, two dimensional (1D, 2D) nuclear magnetic resonance (NMR) methodologies and molecular simulations. NMR investigations revealed that the metal complex intercalates into the DNA base stack from minor groove orientation with dpq as intercalator and dpq ligand participated in the nucleobase stack. Molecular docking simulations of these systems were consistent with the experimental results and revealed that the recognition shows obvious enantioselectivity: the L-isomer is favored at the pyrimidine-purine/purine-pyrimidine region, especially CG/GC sequence, while the D-isomer is favored at the pyrimidine-pyrimidine/purine-purine region, especially TC/AG sequence. Surprisingly, we found that the L-isomer would be enriched when racemic complex was employed. So contrary to general viewpoint, the L-isomer of complex recognized this right hand double helical DNA preferentially. Detailed analysis suggests that it is the electrostatic interactions that determine the steric interactions and then determine the whole recognition event. PMID- 15833337 TI - C-MALISA (cellular magnetic-linked immunosorbent assay), a new application of cellular ELISA for MRI. AB - A modified cellular ELISA (enzyme-linked immunosorbent assay), named cellular magnetic-linked immunosorbent assay (C-MALISA), has been developed as an application of magnetic resonance imaging (MRI) for in vitro clinical diagnosis. To validate the method, three contrast agents targeted to integrins were synthesized by grafting to USPIO (ultrasmall particles of iron oxide): (a) the CS1 (connecting segment-1) fragment of fibronectin (FN) (USPIO-g-FN); (b) the peptide GRGD (USPIO-g-GRGD); (c) a non-peptidic RGD mimetic (USPIO-g-mimRGD). Jurkat cells and rat mononuclear cells were stimulated to activate their integrins. After cell fixation on ELISA plates, incubation with the contrast agents, rinsing, and digestion in 5N HCl, the samples were analyzed by MRI. Paramagnetic relaxation rate enhancements (delta R2) were measured on images. Delta R2 was converted in values of iron concentration based on a calibration curve. The apparent dissociation constants (K(d)*) of the three contrast agents were estimated based on the MRI measurement of delta R2. K(d)* of 1.22 x 10(-7) M, of 7.00 x 10(-8) M, and of 1.13 x 10(-8) M were found respectively for USPIO-g FN, USPIO-g-GRGD, and USPIO-g-mimGRG. The MRI confirmed a statistically significant difference (p < 0.01, p < 0.05) between the stimulated cells incubated with integrin-targeted compounds with respect to the controls (i.e., non-stimulated cells and stimulated cells incubated with non-specific USPIO). The integrin specificity of the three compounds was confirmed by the pre-incubation with GRGD (for USPIO-g-mimRGD and USPIO-g-GRGD) or FN (for USPIO-g-FN). PMID- 15833338 TI - Thermodynamics of the interaction of aluminum ions with DNA: implications for the biological function of aluminum. AB - Aluminum is a known neurotoxic agent and its neurotoxic effects may be due to its binding to DNA. However, the mechanism for the interaction of aluminum ions with DNA is not well understood. Here, we report the application of isothermal titration calorimetry (ITC), fluorescence spectroscopy, and UV spectroscopy to investigate the thermodynamics of the binding of aluminum ions to calf thymus DNA (CT DNA) under various pH and temperature conditions. The binding reaction is driven entirely by a large favorable entropy increase but with an unfavorable enthalpy increase in the pH range of 3.5-5.5 and at all temperatures examined. Aluminum ions show a strong and pH-dependent binding affinity to CT DNA, and a large positive molar heat capacity change for the binding, 1.57 kcal mol(-1) K( 1), demonstrates the burial of the polar surface of CT DNA upon groove binding. The fluorescence of ethidium bromide bound to CT DNA is quenched by aluminum ions in a dynamic way. Both Stern-Volmer quenching constant and the binding constant increase with the increase of the pH values, reaching a maximum at pH 4.5, and decline with further increasing the pH to 5.5. At pH 6.0 and 7.0, aluminum ions precipitate CT DNA completely and no binding of aluminum ions to CT DNA is observed by ITC. Combining the results from these three methods, we conclude that aluminum ions bind to CT DNA with high affinity through groove binding under aluminum toxicity pH conditions and precipitate CT DNA under physiological conditions. PMID- 15833339 TI - Effects of acute and chronic oval exposure of lead on blood pressure and bone mineral density in rats. AB - The immediate and late deleterious effects of early lead (Pb) exposure on human health are major challenges to the health professionals. Therefore, a major underlying contribution of Pb accumulated earlier in life to chronic disorders such as hypertension and osteopenia, commonly found in the elderly, cannot be definitively excluded. Using Sprague-Dawley rats, we examined the effects of drinking 1% Pb acetate on very young rats for 40 days on Pb kinetics, body weight (BW), systolic blood pressure (SBP) and bone mineral density (BMD) over a year long period. We also examined the effects of early and later chelation using disodium calcium ethylenediaminetetraacetate (Na2CaEDTA). The early ingestion and accumulation of Pb was associated with an increased SBP acutely that returned to control levels with continuing challenge and later rose above control months later after the Pb challenge had been halted. Early accumulation of Pb over a short period was also associated with decreased BMD at the end of one year. Chelation after the initial Pb challenge at various times favorably influenced the SBP but could not prevent the lowering of BMD. PMID- 15833340 TI - Spectroscopic investigation of isonitrile complexes of ferric and ferrous microperoxidase 8. AB - Microperoxidase 8 (MP8) is able to react with alkyl- and aryl-isonitriles (RNC) both in its reduced and oxidized states, to form MP8Fe(II)- and MP8Fe(III)-CNR complexes. The coordination and spin states of these complexes have been fully characterized by UV-visible and resonance Raman spectroscopies. Both MP8Fe(II)- and MP8Fe(III)-CNR complexes are hexacoordinate low-spin complexes, which bear a single RNC ligand on the distal face of the heme and keep the His 18 ligand on its proximal face, trans to the RNC ligand. A comparison of these characteristics with those of the Fe-CNR complexes of other hemoproteins suggests that both MP8Fe(II)- and MP8Fe(III)-CNR complexes present a Fe-C-N linear arrangement. This may be due to the lack of any interactions of the RNC ligand with the octapeptide of MP8 that is mainly located over the opposite face of the heme. Finally the formation of hexacoordinate low-spin MP8Fe(II)- and MP8Fe(III)-CNR complexes constitutes a new example of the reactivity of MP8 with a new class of weak sigma donating and strong pi-accepting ligands, which adds to its already very rich coordination chemistry. PMID- 15833341 TI - Beryllium binding at neutral pH: the importance of the Be-O-Be motif. AB - Beryllium speciation at physiological conditions is critical to understanding chronic beryllium disease (CBD). The MHC-class II receptor alleles that have been linked to CBD have more than six carboxylates in a short 20 amino acid segment of the binding pocket and it has been suggested that beryllium may bind within the MHC-class II receptor via the carboxylates. Previous reports also show that citric acid binds beryllium significantly stronger than similar carboxylate ligands such as tartaric acid and is one of the few ligands that can compete with hydrolysis to solubilize beryllium across the entire pH range at molar concentrations. We have characterized the binding of Be to citric acid and shown using a combination of NMR, mass spectrometry and ligand competition studies that Be2L and Be4L2 species dominate. A Be-O-Be linkage with the bridging oxygen coming from the aliphatic alcohol is critical to the stability of the complex. We show through competition experiments that the most stable Be-O-Be arrangement has one Be in a five-member ring and the other Be in a six-member ring. The unusual deprotonation of an aliphatic alcohol (pK(a) = 18) at neutral pH has significant ramifications on the potential interactions of Be with biological ligands such as carbohydrates and Ser and Thr residues. PMID- 15833342 TI - A multi-approach study of the interaction of the Cu(II) and Ni(II) ions with alanylglycylhistamine, a mimicking pseudopeptide of the serum albumine N-terminal residue. AB - The protonation equilibria of alanylglycylhistamine (Ala-Gly-Ha) and the complexation of this ligand with Cu(II) and Ni(II) have been studied by pH potentiometry, 1H and 14N NMR spectroscopy, electrospray ionization mass spectrometry (ESI-MS), circular dichroism (CD), UV-Vis spectrophotometry and electron paramagnetic resonance (EPR). From pH approximately 2-12, the following complexes: MLH, MLH(-1), MLH(-2) and MLH(-3) are successively formed in aqueous solutions, the ligand under its neutral form being noted L. At physiological pH, the MLH(-2) complex is predominant. The coordination in this complex is assumed by one amino, two deprotonated peptide and one imidazole nitrogen atoms. The ESI MS study confirmed the formation of the MLH(-1), MLH(-2) and MLH(-3) complexes. The structure of MLH(-2) was determined by single crystal X-ray analysis. CD and UV-Vis techniques allowed us to propose that the imidazole-N3 nitrogen acts as the anchor group for the coordination to the metal(II) ions rather than the amino group. At high pH values, the further deprotonation of the N-H imidazole group, leading to the formation of MLH(-3), occurs, as revealed by 1H NMR spectroscopy. PMID- 15833343 TI - Partially and fully beta-brominated Mn-porphyrins in P450 biomimetic systems: effects of the degree of bromination on electrochemical and catalytic properties. AB - Beta-hexabromo-5,10,15,20-tetrakis(4 carbomethoxyphenyl)porphyrinatomanganese(III) chloride (Mn(III)(Br6TCMPP)Cl) was prepared by selective Br2-hexabromation of its parent non-brominated manganese complex (Mn(III)(TCMPP)Cl), whereas the octabrominated analogue beta-octabromo 5,10,15,20-tetrakis(4-carbomethoxyphenyl)porphyrinatomanganese(III) chloride (Mn(III)(Br8TCMPP)Cl) was synthesized via metallation of the corresponding free base. Beta-octabromo-5,10,15,20-tetrakis(4-carbomethoxyphenyl)porphyrin was obtained by demetallation of its brominated Cu(II) derivative, which, in its turn, was prepared by either a Br2 or an N-bromosuccinimide protocol. Relative to Mn(III)(TCMPP)Cl (E(1/2) = -0.16 V vs. normal hydrogen electrode, CH2Cl2), the Mn(III)/Mn(II) reduction potential of Mn(III)(Br8TCMPP)Cl and Mn(III)(Br6TCMPP)Cl showed anodic shifts of 0.43 and 0.33 V, respectively, which corresponded to a linear shift of 0.05 V per bromine added. These manganese complexes were evaluated as cytochrome P450 mimics in catalytic iodosylbenzene (PhIO)-oxidations of cyclohexane and cyclohexene. In aerobic PhIO-oxidation of cyclohexene, epoxidation and allylic autoxidation reactions were inversely related, competitive processes; the most efficient P450-mimics were the least effective autoxidation catalysts. Mn(III)(Br6TCMPP)Cl was more efficient as epoxidation or hydroxylation catalyst than both its fully and non-beta-brominated counterparts were. There was no linear relationship between the catalytic efficiency and both the number of bromine substituents and the Mn(III)/Mn(II) potential; these observations were compared to Lyons system literature data and discussed. Analogously to enzymatic optimum pH effects, an optimum redox potential effect is suggested as relevant in designing and understanding cytochrome P450 biomimetic catalysts. PMID- 15833344 TI - Copper(II) complexes of 1,10-phenanthroline-derived ligands: studies on DNA binding properties and nuclease activity. AB - A series of copper(II) complexes of the type [Cu(L)]2+, where L = N,N'-dialkyl 1,10-phenanthroline-2,9-dimethanamine and R = methyl (L1), n-propyl (L2), isopropyl (L3), sec-butyl (L4), or tert-butyl (L5) group, have been synthesized. The interaction of the complexes with DNA has been studied by DNA fiber electron paramagnetic resonance (EPR) spectroscopy, emission, viscosity and electrochemical measurements and agarose gel electrophoresis. In the X-ray crystal structure of [Cu(HL2)Cl2]NO3, copper(II) is coordinated to two ring nitrogens and one of the two secondary amine nitrogens of the side chains and two chloride ions as well and the coordination geometry is best described as trigonal bipyramidal distorted square based pyramidal (TBDSBP). Electronic and EPR spectral studies reveal that all the complexes in aqueous solution around pH 7 possess CuN3O2 rather than CuN4O chromophore with one of the alkylamino side chain not involved in coordination. The structures of the complexes in aqueous solution around pH 7 change from distorted tetragonal to trigonal bipyramidal as the size of the alkyl group is increased. The observed changes in the physicochemical features of the complexes on binding to DNA suggest that the complexes, except [Cu(L5)]2+, bind to DNA with partial intercalation of the derivatised phen ring in between the DNA base pairs. Electrochemical studies reveal that the complexes prefer to bind to DNA in Cu(II) rather than Cu(I) oxidation state. Interestingly, [Cu(L5)]2+ shows the highest DNA cleavage activity among all the present copper(II) complexes suggesting that the bulky N tert-butyl group plays an important role in modifying the coordination environment around the copper(II) center, the Cu(II)/Cu(I) redox potential and hence the formation of activated oxidant responsible for the cleavage. These results were compared with those for bis(1,10-phenanthroline)copper(II), [Cu(phen)2]2+. PMID- 15833345 TI - Standard reduction potentials of all couples of the peroxidase cycle of lactoperoxidase. AB - Lactoperoxidase (LPO) is found in mucosal surfaces and exocrine secretions including milk, tears and saliva and has physiological significance in antimicrobial defense. Its predominant physiological role is to convert hydrogen peroxide and thiocyanate in hypothiocyanite. In this study, the standard reduction potentials of all redox couples involved in the halogenation and peroxidase cycle of LPO have been determined by multi-mixing stopped-flow spectroscopy. The standard reduction potentials of the redox couples compound I/native LPO, compound I/compound II of LPO, and compound II/native LPO are (1.09 +/- 0.01) V, (1.14 +/- 0.02) V, and (1.04 +/- 0.02) V, respectively, at pH 7 and 25 degrees C. Thus, for the first time, a full description of these important thermodynamic parameters of lactoperoxidase has been performed, allowing a better understanding in the substantial differences in the oxidation of two- and one electron donors by LPO and other members of the mammalian heme peroxidase superfamily. PMID- 15833346 TI - Biphenyl derived oxovanadium(IV) and copper(II) salen-type complexes--structure and redox tuning. AB - A series of vanadyl(IV) salen (N,N'-bis(salicylidene)ethylenediaminato)-type complexes (1-4) bearing phenyl or 2-hydroxyphenyl moieties have been prepared and characterized by means of mass spectrometry, infra-red, electron paramagnetic resonance (EPR), UV/Vis spectroscopy, cyclovoltammetry and X-ray crystallography. Their structures have been compared to their copper(II) analogs 5-8. Hydrogen intralinkages have been observed in the crystal structure of 5. The pendant hydroxy groups fine-tune the redox properties of the complexes. The catalytic activity in the oxygenation of ethyl phenyl sulfide to the corresponding sulfoxide was investigated. Results indicate that complex 1 bearing hydroxyphenyl subunits and a phenylene bridge is the most selective under these reaction conditions, with the smallest amount of the over-oxidized product, sulfone. PMID- 15833347 TI - Vanadate oligomers: in vivo effects in hepatic vanadium accumulation and stress markers. AB - The formation of vanadate oligomeric species is often disregarded in studies on vanadate effects in biological systems, particularly in vivo, even though they may interact with high affinity with many proteins. We report the effects in fish hepatic tissue of an acute intravenous exposure (12, 24 h and 7 days) to two vanadium(V) solutions, metavanadate and decavanadate, containing different vanadate oligomers administered at sub-lethal concentration (5 mM; 1 mg/kg). Decavanadate solution promotes a 5-fold increase (0.135 +/- 0.053 microg V(-1) dry tissues) in the vanadium content of the mitochondrial fraction 7 days after exposition, whereas no effects were observed after metavanadate solution administration. Reduced glutathione (GSH) levels did not change and the overall reactive oxygen species (ROS) production was decreased by 30% 24 h after decavanadate administration, while for metavanadate, GSH levels increased 35%, the overall ROS production was depressed by 40% and mitochondrial superoxide anion production decreased 45%. Decavanadate intoxication did not induce changes in the rate of lipid peroxidation till 12 h, but later increased 80%, which is similar to the increase observed for metavanadate after 24 h. Decameric vanadate administration clearly induces different effects than the other vanadate oligomeric species, pointing out the importance of taking into account the different vanadate oligomers in the evaluation of vanadium(V) effects in biological systems. PMID- 15833348 TI - Direct electrochemistry of engineered cytochrome b562 molecules with a ligand binding pocket. AB - The rapid and reversible electron transfer reaction of cytochrome b562 was observed at an In2O3 electrode. The estimated heterogeneous electron transfer rate constant (k0') was k0' > or = 5.0 x 10(-3) cm s(-1) at pH 6.5. When the methionine-7 (Met-7) residue, which coordinates to the heme iron as an axial ligand, of the wild-type cytochrome b562 was replaced by an Ala or Gly residue, a water molecule bound to the heme iron and the electron transfer rate constants decreased to 1.3 x 10(-3) and 1.8 x 10(-3) cm s(-1), respectively. This decrease in the electron transfer rate would be due to the larger reorganization energy for the structural change at the redox site. The midpoint potential of cytochrome b562 was shifted negatively by approximately 135 mV by replacing Met-7 with Ala or Gly. Similar dissociation kinetics of cyanide for the mutated molecules as compared to native myoglobin was obtained. PMID- 15833349 TI - Structural and spectroscopic characterization of aqua-diargininate-copper(II) carbonate monohydrate. AB - The molecular structure of the title complex, [Cu(C6H14N4O2)2(H2O)]CO3.H(2)O, was determined by single crystal X-ray diffractometry. It crystallizes in the monoclinic space group P2(1), with Z = 2. The Cu(II) ion is in a square pyramidal environment, trans coordinated at the basis by two argininate groups acting as bidentate ligands through their amino nitrogen and carboxylate oxygen atoms. The coordination around copper is completed by a water molecule at the pyramid apex. The infrared, Raman and electronic spectra are briefly discussed on the basis of the structural peculiarities of the complex. PMID- 15833350 TI - Proton-linked subunit heterogeneity in ferrous nitrosylated human adult hemoglobin: an EPR study. AB - The effect of pH on the X-band electron paramagnetic resonance (EPR) spectrum of ferrous nitrosylated human adult tetrameric hemoglobin (HbNO) as well as of ferrous nitrosylated monomeric alpha- and beta-chains has been investigated, at 163 degrees C. At pH 7.3, the X-band EPR spectrum of tetrameric HbNO and ferrous nitrosylated monomeric alpha- and beta-chains displays a rhombic shape. Lowering the pH from 7.3 to 3.0, tetrameric HbNO and ferrous nitrosylated monomeric alpha- and beta-chains undergo a transition towards a species characterized by a X-band EPR spectrum with a three-line splitting centered at 334mT. These pH-dependent spectroscopic changes may be taken as indicative of the cleavage, or the severe weakening, of the proximal HisF8-Fe bond. In tetrameric HbNO, the pH-dependent spectroscopic changes depend on the acid-base equilibrium of two apparent ionizing groups with pK(a) values of 5.8 and 3.8. By contrast, the pH-dependent spectroscopic changes occurring in ferrous nitrosylated monomeric alpha- and beta chains depend on the acid-base equilibrium of one apparent ionizing group with pK(a) values of 4.8 and 4.7, respectively. The different pK(a) values for the proton-linked spectroscopic transition(s) of tetrameric HbNO and ferrous nitrosylated monomeric alpha- and beta-chains suggest that the quaternary assembly drastically affects the strength of the proximal HisF8-Fe bond in both subunits. This probably reflects a 'quaternary effect', i.e., structural changes in both subunits upon tetrameric assembly, which is associated to a relevant variation of functional properties (i.e., proton affinity). PMID- 15833351 TI - Secretion of an aluminum chelator, 2-isopropylmalic acid, by the budding yeast, Saccharomyces cerevisiae. AB - An aluminum(III)-binding substance (ABS), that solubilizes Al(III) at neutral pH, was found to be secreted by Saccharomyces cerevisiae. A combination of anion exchange chromatography and preparative high performance liquid chromatography using an octadecylsilane (ODS) column separated ABS from the medium. The structure determination of ABS was performed using 1H and 13C NMR spectroscopy and heteronuclear multiple-bond correlation (HMBC) spectroscopy, and ABS was identified to be 2-isopropylmalic acid (2-iPMA). The structure was further confirmed using high resolution electrospray ionization mass spectrometry. Solubilization of otherwise sparingly soluble Al(III) by 2-iPMA at neutral pH indicated the binding of the compound with Al(III). This is supported by 27Al NMR spectrometry for a solution containing 10 mM Al(III) and 20 mM 2-iPMA at pH 6.6, where four Al(III) species were evident. Although the function of this compound is unclear, it might play a key role in Al detoxification. PMID- 15833352 TI - The first enantioselective synthesis of the amavadin ligand and its complexation to vanadium. AB - The ligand of the naturally occurring vanadium compound amavadin found in Amanita muscaria, (2S, 2'S)-N-hydroxyimino-2,2'-dipropionic acid (1), was synthesized stereoselectively in two steps with 43% overall yield. After complexation of this ligand to vanadyl acetate, amavadin was isolated in quantitative yield. Due to the chirality at vanadium amavadin consists of a mixture of delta and lambda diastereoisomers. Directly after its synthesis, the delta to lambda ratio of amavadin is 2.27 and it decreases to 0.80 after equilibrium has been reached. During this epimerization the optical rotation for V[(2S,2'S)-N-hydroxyimino (2,2')-dipropionate]2 (=amavadin) changes from [alpha](D)25 = +36 degrees to +114.0 degrees (c = 0.5, H2O). For V[(2R,2'R)-N-hydroxyimino-(2,2')-dipropionate] the optical rotation changes from [alpha](D)25 = -36 degrees to -113.2 degrees (c = 0.5, H2O). PMID- 15833353 TI - Serum or plasma: what kind of blood sample should be used to measure circulating matrix metalloproteinases and their inhibitors? PMID- 15833354 TI - Cis acting expression loci in multiple sclerosis. PMID- 15833355 TI - Characterization of light chain immunoglobulin in urine from animals and humans infected with prion diseases. AB - The necessity of a non-invasive in-vivo test for prion diseases has become more apparent since the transmission of vCJD from the blood of a healthy individual incubating the disease. Here we show that prion urine comprises an array of protease resistant peptides, among them light chain immunoglobulin (LC). This was observed by sequencing gel bands comprising hamster urine samples, as well as by immunoblotting of similar samples with anti mouse IgG reagents for hamster samples, or with anti human IgG reagents for human samples. Our result suggests that urine samples from CJD patients can be identified by the presence of protease resistant proteins such as LC. PMID- 15833356 TI - Choroid plexus selectively accumulates T-lymphocytes in normal controls and after peripheral immune activation. AB - We determined T-lymphocyte migration into brain and choroid plexus (CPx) after enterotoxin-induced systemic immune activation. CPx T-lymphocytes/mm2 in control mice were > 3 logs more numerous than brain and increased by as much as 150-fold by post-enterotoxin Day 3 (p < 0.01). Flow cytometry of pooled CPx confirmed post enterotoxin increases. Brain T-lymphocytes increased up to 17-fold after SEB and accumulated in subependymal and periventricular brain. T cell apoptosis was absent. These results show preferential T-lymphocyte migration to CPx over brain and suggest that brain T cells may be derived from the CPx by direct migration or by cerebrospinal fluid dissemination. PMID- 15833357 TI - Death receptor-mediated apoptosis in human malignant glioma cells: modulation by the CD40/CD40L system. AB - CD40, a TNF-R-related cell surface receptor, is shown here to be expressed by glioma cells in vitro and in vivo. Glioma cell lines expressing low levels of CD40 at the cell surface resist cytotoxic effects of CD40L. CD40 gene transfer sensitizes glioma cells to CD40L. Inhibition of protein synthesis potentiates cell death which involves CD40 clustering and caspases 8 and 3 processing. CD40 transfected LN-18 cells acquire resistance to CD95L. In contrast, subtoxic concentrations of CD40L strongly sensitize these cells for TNF-alpha-induced apoptosis. Bispecific CD40xCD95 antibodies specifically kill glioma cells, disclosing the property of endogenous CD40 to facilitate death signalling. PMID- 15833358 TI - The immunomodulating neuropeptide alpha-melanocyte-stimulating hormone (alpha MSH) suppresses LPS-stimulated TLR4 with IRAK-M in macrophages. AB - Since alpha-MSH suppresses endotoxin-induced inflammation by innate immunity, it is possible that alpha-MSH can suppress the interface between innate and adaptive immunity mediated by TLR4-stimulated macrophages. Endotoxin-stimulated macrophages treated with alpha-MSH are suppressed in nitric oxide and IL-12p70 production, and cannot enhance antigen-stimulated IFN-gamma production by Th1 cells. In macrophages treated with alpha-MSH, the inhibitory molecule IRAK-M is bound to IRAK-1, the proximal intracellular signal molecule of endotoxin-bound TLR4. These results further demonstrate the dynamic contribution of the nervous system, and the role of alpha-MSH in modulating the innate and adaptive immune interface in an inflammatory response. PMID- 15833359 TI - Chemokine receptor deficiency is associated with increased chemokine expression in the peripheral and central nervous systems and increased resistance to herpetic encephalitis. AB - Herpes simplex virus type 1 (HSV-1) infection of the eye leads to the retrograde spread of the virus from the eye to the trigeminal ganglion resulting in the infiltration of leukocytes and production of inflammatory cytokines and chemokines including CXCL9 and CXCL10. The present study investigated the role of the receptor for CXCL9 and CXCL10 in the host response to HSV-1 infection using mice deficient in CXCR3 expression (CXCR3-/-). Although wild type C57BL/6 and CXCR3-/- mice cleared the virus, HSV-1 titers remained elevated in the ganglion and brain stem of CXCR3-/- mice day 7 post infection. Coinciding with the increase in virus titer, CCL5, CXCL9, CXCL10 and IFN-gamma protein levels were enhanced in the trigeminal ganglion and/or brain stem of the CXCR3-/- mice associated with a 2-fold increase in the percentage of CD3+CD8+ T lymphocytes in the trigeminal ganglion. However, the survival rate of CXCR3-/- mice was significantly enhanced above the wild type controls associated with an increase in brain IL-6 content. Collectively, the results indicate the absence of CXCR3 is associated with a transient increase in virus burden in the nervous system and an elevated protective immune response. PMID- 15833360 TI - Transcriptome profile within the mouse central nervous system and activation of myelin-reactive T cells following murine coronavirus infection. AB - Multiple sclerosis (MS) is an autoimmune disease associated with environmental factors, possibly including several viruses such as the coronaviruses. Indeed, murine coronavirus (MHV) infection provides a well-known experimental model for MS studies. Intracerebral infection of C57BL/6 mice with MHV-A59 revealed that viral replication was efficient and that clearance of infectious virus occurred as soon as 7 days post-infection. Using cDNA arrays, analysis of gene expression profile in the brain revealed a modulation of 80 different genes following infection, with at least 27 of these genes having previously been directly related to innate or acquired immune responses. Concordingly, an important activation of auto-reactive T cells specific to myelin basic protein was demonstrated. Altogether, these results indicate that an MHV infection of the central nervous system (CNS) leads to an important host genomic response implicating immunity-related genes and to the activation of myelin-reactive autoimmune T cells. PMID- 15833361 TI - Interleukin-10 inhibits endotoxin-induced pro-inflammatory cytokines in microglial cell cultures. AB - Inflammation contributes to perinatal brain injury and can be induced by hypoxia ischemia (HI) or exposure to infection (fetal inflammatory response). The anti inflammatory cytokine interleukin-10 (IL10) has been shown to have neuroprotective effects following HI. To determine whether IL10 can reduce the inflammatory response to lipopolysaccharide (LPS) in microglial cell cultures, primary microglial (MG) and/or HAPI cells (new MG-like cell line) were treated with LPS (50 ng/ml) in the presence or absence of IL10 (20 ng/ml) for 0.5, 1, 4, and 8 h. TNFalpha, MIP-1alpha, and RANTES were assayed by ELISA. Chemokine receptors, CCR5, CXCR3, and CX3CR1 (fractalkine receptor) were assayed by semiquantitative RT-PCR. We found that in MG cell cultures TNFalpha, MIP-1alpha, and RANTES release after 8-h exposure to LPS was significantly higher compared to non-exposed MG cells (P < 0.001). In HAPI cell cultures similar stimulation of mRNA levels was found for TNFalpha, MIP-1alpha, CXCR3, and CX3CR1. IL10 inhibited TNFalpha, MIP-1alpha, and RANTES release of LPS-stimulated MG cells as well as TNFalpha, MIP-1alpha, and CXCR3 mRNA expression by HAPI cells after exposure to LPS (P < 0.05). In contrast to those inhibitory effects, there was no change in fractalkine, and a modest increase in CX3CR1 mRNA levels was found in the presence of IL10. We conclude that the inflammatory response induced in microglial cells by LPS can be markedly reduced by IL10. The increase in fractalkine receptor (CX3CR1) is also potentially protective. Our results suggest that treatment of damaging neuroinflammatory insults such hypoxia-ischemia, with IL10 may be protective for the immature brain. PMID- 15833362 TI - Pregnancy and social stress in female rats: influences on blood leukocytes and corticosterone concentrations. AB - The consequences of pregnancy and social stress on blood immune cells and on plasma corticosterone concentrations were assessed in Long Evans rats. Normal pregnancy in control females was characterized by a progressive increase in corticosterone concentration and increasing numbers of granulocytes. In contrast, CD4 T, CD8 T, and B cell numbers as well as the proliferative response of lymphocytes decreased as pregnancy progressed. Stress was induced in pregnant females by social confrontation for 2 h daily with a female resident opponent over a period of 2 months. Corticosterone concentrations were substantially higher in pregnant stressed than in pregnant control rats. Furthermore, the numbers of monocytes, NK and B cells were lower in stressed females, and there was a strong trend towards suppressed lymphocyte proliferation. Interestingly, pregnant females did not show granulocytosis in response to the stressor. In sum, the social stress paradigm in females appears to be a good model for the investigation of the interactions between stress, pregnancy and the immune system. It also provides an excellent platform for studies on prenatal stress under relatively naturalistic conditions. PMID- 15833363 TI - Autocrine activation of microglia by tumor necrosis factor-alpha. AB - In the central nervous system (CNS), tumor necrosis factor-alpha (TNF-alpha) derived from activated microglia plays a critical role as an inflammatory mediator. In this study, we examined the function of TNF-alpha as an autocrine mediator in microglial activation. TNF-alpha induced TNF-alpha production by microglia through ligation of TNF receptor 1 (TNFR1). TNF-alpha also increased the production of other inflammatory mediators. The activation of microglia by lipopolysaccharide is partially mediated by microglia-derived TNF-alpha. These findings suggest the existence of a positive feedback loop in the activation of microglia via TNF-alpha. This autocrine loop may be involved in the prolonged activation of microglia. PMID- 15833364 TI - CRH stimulates POMC activity and corticosterone production in dermal fibroblasts. AB - It has been previously documented that human skin cells including epidermal keratinocytes and dermal fibroblasts produce and process proopiomelanocortin (POMC), corticotropin releasing hormone (CRH), and express functional CRH receptors type-1 (CRH-R1). The skin also has corticosteroidogenic activity, suggesting a functional connection between these elements. In the current study, we found that human dermal fibroblasts (but not normal epidermal keratinocytes) respond to CRH with stimulation of cAMP, with POMC gene and protein expression, and ACTH production and release. Furthermore, CRH and ACTH stimulate production of corticosterone in fibroblasts, with ACTH being more potent. Although cortisol immunoreactivity accumulation/production in fibroblasts has been detected by ELISA, it appears to be constitutive (not affected by CRH or ACTH). These effects are absent in keratinocytes. Therefore, we propose that fibroblasts but not keratinocytes display a functional CRH-POMC-corticosteroid axis organized similarly to the hypothalamus-pituitary-adrenal (HPA) axis. However, it diverges from the HPA organization in its distal step, where CRH and ACTH stimulate production of corticosterone, instead of cortisol. PMID- 15833365 TI - Restraint stress alters lung gene expression in an experimental influenza A viral infection. AB - In the present study the global effect of restraint stress on gene expression in the murine lung during an experimental influenza A/PR8 viral infection was examined. Gene expression profiling using high density oligonucleotide microarrays revealed that the expression of 95 genes was altered on day 3 post infection (p.i.), while 48 genes were altered on day 7 p.i. Restraint stress reduced and delayed the expression of specific cytokines, cell adhesion molecules and cell surface receptors indicating alterations in cell migration to the site of infection. Furthermore, mapping of the candidate genes to known pathways revealed that genes associated with host defense and immune responses, including chemotaxis and chemokine function, antigen presentation and processing, MHC class II receptor function and inflammation were the major pathways affected by restraint stress. PMID- 15833366 TI - Interferon-gamma and interferon-beta affect endogenous catecholamines in human peripheral blood mononuclear cells: implications for multiple sclerosis. AB - Interferon (IFN)-gamma plays a pivotal role in the pathogenesis of multiple sclerosis (MS), while IFN-beta may be able to modify the clinical course of the disease, eventually also by counterbalancing IFN-gamma-mediated effects. Catecholamines (CA) exert important effects on the immune response, both as transmitters between the nervous and the immune system, as well as autocrine/paracrine mediators in immune cells, and several lines of evidence support their involvement in MS. In particular, dysregulated production of CA seems to occur in peripheral blood mononuclear cells (PBMCs) of MS patients. We assessed the effects of IFN-beta and IFN-gamma on endogenous CA in PBMCs. In cultured PBMCs stimulated with phytohaemagglutinin (PHA), IFN-beta increased CA production and induced CA release in the culture medium, while IFN-gamma decreased both CA production and the expression of mRNA for the CA-synthesizing enzyme tyrosine hydroxylase. Coincubation with both IFNs prevented the inhibitory effect of IFN-gamma, as well as the stimulatory effect of IFN-beta. IFNs are the first physiological compounds shown to affect endogenous CA in PBMCs: in view of the role of CA-dependent mechanisms in the immune response, these findings may help to better understand the mechanisms of action of IFN-beta as an immunomodulatory drug in MS. PMID- 15833367 TI - CCL2 and CCL5 mediate leukocyte adhesion in experimental autoimmune encephalomyelitis--an intravital microscopy study. AB - Experimental autoimmune encephalomyelitis (EAE) models multiple sclerosis (MS) and is characterized by marked mononuclear cell influx in the brain. Several studies have demonstrated a role for chemokines during EAE. It remains to be determined whether these mediators modulate EAE primarily by mediating leukocyte influx into the CNS or by modifying lymphocyte activation and/or trafficking into lymphoid organs. After induction of EAE with MOG(35-55), leukocyte recruitment peaked on day 14 and correlated with symptom onset, TNF-alpha production and production of CCL2 and CCL5. Levels of CXCL-10 and CCL3 were not different from control animals. Using intravital microscopy, we demonstrated that leukocyte rolling and adhesion also peaked at day 14. Treatment with anti-CCL2 or anti-CCL5 antibodies just prior to the intravital microscopy prevented leukocyte adhesion, but not rolling. Our data suggest that induction of leukocyte adhesion to the brain microvasculature is an important mechanism by which CCL2 and CCL5 participate in the pathophysiology of EAE. PMID- 15833368 TI - Autoantibodies against the amino terminal of alpha-enolase are a useful diagnostic marker of Hashimoto's encephalopathy. AB - We investigated autoantibodies and their epitope(s) in Hashimoto's encephalopathy associated with Hashimoto's thyroiditis. In a proteomic analysis, they proved to recognize alpha-enolase. We further searched the epitope region in alpha-enolase using different regions of recombinant proteins expressed in cultured human cells. The amino terminal region was recognized by autobodies from a much higher proportion of patients with Hashimoto's encephalopathy (83.3%; 5/6) than from patients with Hashimoto's thyroiditis (11.8%; 2/17), and not at all by sera from controls (25 healthy individuals and 25 controls with other neurological disorders) (0%; 0/50). Neither the carboxyl terminal nor the mid-region of alpha enolase showed specificity for Hashimoto's encephalopathy. Autoantibodies against the amino terminal of alpha-enolase are a useful diagnostic marker for Hashimoto's encephalopathy. PMID- 15833369 TI - In vivo corticosterone administration at levels occurring with intense exercise does not induce intestinal lymphocyte apoptosis in mice. AB - Intestinal lymphocyte apoptosis can occur following physiological and pathophysiological stress as well as exhaustive exercise. In this study we investigated whether corticosterone (CORT) administration at physiological concentrations observed following strenuous exercise induces intestinal lymphocyte apoptosis and cell loss in mice. CORT injection (14 mg/kg; i.p.) caused a four-fold increase in plasma CORT concentrations, but did not affect intestinal lymphocyte cell loss or alter baseline intestinal lymphocyte apoptosis, as measured by phosphatidylserine externalization, cell viability, mitochondrial membrane depolarization, caspase 3, Bcl-2 and cytosolic cytochrome c protein levels. These findings indicate that CORT at levels observed following strenuous exercise is not involved in intestinal lymphocyte apoptosis and cell loss. PMID- 15833370 TI - Inhibition of acetylcholine receptor function by seronegative myasthenia gravis non-IgG factor correlates with desensitisation. AB - 15% of myasthenia gravis (MG) patients do not have antibodies against the acetylcholine receptor (AChR). Some of these "seronegative" MG patients have antibodies against muscle specific kinase (MuSK), and many have a non-IgG factor that acutely inhibits AChR function in a muscle-like cell line, CN21. Here we show, using mainly one plasma negative for both AChR and MuSK antibodies, that the inhibitory effect of the non-IgG fraction correlates well with the desensitisation caused by 100 microM nicotine, and is found also when AChRs are expressed in a non-muscle cell line (HEK). Moreover, a similar effect was seen with M3C7-a monoclonal antibody against human AChR. The results suggest that, rather than acting indirectly as previously proposed, the SNMG factor may bind directly to an allosteric site that induces or enhances AChR desensitisation. PMID- 15833371 TI - Severity of Guillain-Barre syndrome is associated with Fc gamma Receptor III polymorphisms. AB - Macrophages and ganglioside-specific IgG are involved in the pathogenesis of Guillain-Barre syndrome (GBS). Leukocyte IgG receptors (Fc gammaR) confer potent cellular effector functions to the specificity of IgG. The efficacy of IgG mediated cellular inflammatory responses is determined by functional polymorphisms of three Fc gammaR subclasses (Fc gammaRIIa: H131/R131; Fc gammaRIIIa: V158/F158; Fc gammaRIIIb: NA1/NA2). Fc gammaR genotype distributions were determined in a Dutch, and British cohort of GBS patients and controls. In addition, a meta-analysis incorporating all previously published data, encompassing a total of 345 GBS patients and 714 healthy controls, was performed. Results suggest that Fc gammaRIII genotypes may represent mild disease-modifying factors in GBS. PMID- 15833372 TI - Systemic T-cell activation in acute clinically isolated optic neuritis. AB - We examined untreated 60 patients with acute monosymptomatic optic neuritis (ON). Patients examined early after onset showed increased expression of HLA-DR and CD45R0 on CD4 and CD8 T cells. Expression of HLA-DR on CD4 T cells was higher in patients without IgG oligoclonal bands. Expression of HLA-DR on CD4 and CD8 T cells correlated negatively with measures of disease activity and positively with measures of good visual function, and expression of CD45R0 on CD4 T cells correlated negatively with measures of disease activity. We hypothesize that HLA DR expression may characterize a protective T-cell subset in ON. PMID- 15833373 TI - Mitoxantrone as rescue therapy in worsening relapsing-remitting MS patients receiving IFN-beta. AB - We assessed the action of mitoxantrone (MX) when given as rescue therapy in patients with relapsing-remitting (RR) multiple sclerosis (MS), whose disease activity worsens despite IFN-beta treatment. Ten very active RR MS patients received MX 12 mg/m2 monthly, for 3 months, and then returned to the original treatment with IFN-beta. Following treatment with MX, 70% of patients were able to return to IFN-beta treatment, stabilising EDSS and relapse rate during a follow-up period of 15-18 additional months. In contrast, in 30% of the patients who were taken off MX and returned to IFN-beta treatment the EDSS score deteriorated and the number of exacerbations increased significantly. The latter patients were switched again to MX treatment at 3-month intervals, stabilising EDSS and relapse rate during 15-18 additional months. Clinical findings correlated with the number of Gd-enhancing lesions disclosed in MRI scans. Immunological data were consistent with the clinical and MRI benefits observed. We conclude that brief courses of MX may provide a safe treatment alternative for RR MS patients who experience rapid and severe worsening of their disease despite IFN-beta treatment. PMID- 15833374 TI - Immune responses to Campylobacter and serum autoantibodies in patients with complex regional pain syndrome. AB - We hypothesised that some complex regional pain syndromes (CRPS) may have a postinfectious and/or autoimmune basis. Sera from 92 patients with CRPS and 92 controls were investigated for immunoreactivity to Campylobacter strains and to rodent tissues. Both IgA-antibodies to Campylobacter and tissue-specific reactivity were often present in patients with short disease duration (< or = 1.5 years). Patients with minimal preceding trauma had stronger nervous tissue specific reactivity than other patients, regardless of disease duration. These results provide preliminary evidence for immune activation early in CRPS and, additionally, that patients with minimal trauma may comprise an autoimmune subgroup. PMID- 15833375 TI - Modulation of prolactin expression in human T lymphocytes by cytokines. AB - Besides its pivotal role in reproduction, the polypeptide hormone prolactin (PRL) has immunomodulatory properties. Whereas the bulk of circulating PRL is produced by the pituitary, PRL is also produced by the decidua, the myometrium, the mammary gland and leukocytes. Extrapituitary PRL expression is regulated differently from that in the pituitary, due to the use of an alternative promoter. Here we show for the first time that in T lymphocytes PRL expression is subject to regulation by cytokines. We established that both IL-2 and IL-4 reduced PRL mRNA levels in T lymphocytes to 25 and 28% of control values, respectively. PRL mRNA expression was inhibited to a lesser extent by IL-1beta, which decreased PRL mRNA levels to 58% of control values. PMID- 15833376 TI - Safety assessment of esters of p-hydroxybenzoic acid (parabens). AB - Parabens are widely used as preservatives in food, cosmetic and pharmaceutical products. Acute, subchronic, and chronic studies in rodents indicate that parabens are practically non-toxic. Parabens are rapidly absorbed, metabolized, and excreted. In individuals with normal skin, parabens are, for the most part, non-irritating and non-sensitizing. However, application of compounds containing parabens to damaged or broken skin has resulted in sensitization. Genotoxicity testing of parabens in a variety of in vitro and in vivo studies primarily gave negative results. The paraben structure is not indicative of carcinogenic potential, and experimental studies support these observations. Some animal studies have reported adverse reproductive effects of parabens. In an uterotrophic assay, methyl and butyl paraben administered orally to immature rats were inactive, while subcutaneous administration of butyl paraben produced a weak positive response. The ability of parabens to transactivate the estrogen receptor in vitro increases with alkyl group size. The detection of parabens in a small number of breast tumor tissue samples and adverse reproductive effects of parabens in animals has provoked controversy over the continued use of these substances. However, the possible estrogenic hazard of parabens on the basis of the available studies is equivocal, and fails to consider the metabolism and elimination rates of parabens, which are dose, route, and species dependent. In light of the recent controversy over the estrogenic potential of parabens, conduct of a reproductive toxicity study may be warranted. PMID- 15833377 TI - Evaluation of estrogenic activities of hydroxylated polycyclic aromatic hydrocarbons in cigarette smoke condensate. AB - Estrogenic activities of cigarette smoke condensates obtained from the extraction of particulate matters from mainstream and sidestream cigarette smoke with benzene/ethanol were evaluated by using a yeast two-hybrid assay system expressing human estrogen receptor alpha (hERalpha). To identify the constituents of the cigarette smoke condensate which are responsible for the estrogenic activity, the condensate was fractionated into eleven fractions by liquid-liquid extractions. Among these fractions, the neutral fractions of mainstream and sidestream smoke showed the strongest estrogen receptor-mediated activity by the yeast two-hybrid assay. Then the neutral fractions were fractionated by medium pressure liquid chromatography with silica gel column. In the fractions that showed strong estrogenic activity, 2-hydroxyfluorene (2-OHFle), 2- and 3 hydroxyphenanthrene (2- and 3-OHPhe), 1-hydroxypyrene (1-OHPyr) and n-propyl-p hydroxybenzoate (n-PHB) were identified by LC- and GC-MS and HPLC with fluorescence detection. 2-OHFle, 2-OHPhe and n-PHB exhibited estrogenic activity, whereas weak activity was observed with 3-OHPhe and 1-OHPyr. Several other hydroxylated polycyclic aromatic hydrocarbons having no activity were also identified. This is a first study to identify estrogenic hydroxylated PAHs in cigarette smoke condensate. The present findings points out the necessity for detailed investigation of exposure to aerosols containing apparently estrogenic compounds. PMID- 15833378 TI - Diallyl disulfide inhibits N-acetyltransferase activity and gene expression in human esophagus epidermoid carcinoma CE 81T/VGH cells. AB - Individuals can be classified into rapid or slow acetylators based on the N acetyltransferase (NAT) activity which is believed to affect cancer risk that is related to environmental carcinogen exposure. Diallyl disulfide (DADS) is a naturally occurring organosulfur compound, from garlic (Allium sativum), which exerts anti-neoplasm activity. In this study, we investigated the inhibitory effects of DADS on NAT activity and gene expresseion (NAT mRNA) in human esophagus epidermoid carcinoma CE 81T/VGH cells. NAT activity was measured by the amounts of N-acetylation of 2-aminofluorene (AF) and non-acetylation of AF by high performance liquid chromatography on cells treated with or without DADS. The amounts of NAT enzymes were examined and analyzed by Western blot. NAT gene expression (NAT mRNA) was examined by polymerase chain reaction and cDNA microarray. DADS decreased the amount of N-acetylation of AF in human esophagus epidermoid carcinoma CE 81T/VGH cells in a dose-dependent manner. Western blot analysis indicated that DADS decreased the levels of NAT protein in CE 81T/VGH cells. PCR and cDNA microarray experiments showed that DADS affected NAT1 mRNA expression in CE 81T/VGH cells. DADS affect NAT activity due to the inhibition of gene expression (NAT1 mRNA) and the decreasing of the protein levels of NAT in CE 81T/VGH cells. PMID- 15833379 TI - Delayed effects of thallium in the rat brain: regional changes in lipid peroxidation and behavioral markers, but moderate alterations in antioxidants, after a single administration. AB - Thallium (Tl+) toxicity has been related with the generation of reactive oxygen species (ROS) and oxidative stress (OS) in the central nervous system. Since changes in endogenous antioxidant systems might contribute to acute Tl+-induced OS and neurotoxicity, in this study we measured the metal concentration and the levels of lipid peroxidation (LP) in different brain regions (hypothalamus (Ht); cerebellum (Ce); striatum (S); hippocampus (Hc) and frontal cortex (Cx)) in possible correlation with the content of reduced glutathione (GSH), the activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD), and the animal performance in behavioral tests, all evaluated after a single administration of thallium acetate (8 or 16 mg/kg, i.p.) to rats. Seven days after Tl+ administration, the metal was homogeneously and dose-dependently accumulated in all regions evaluated. LP was increased in Ht, Ce and S, while GSH was depleted in S. Cu,Zn-SOD activity was also decreased in Ht and S. All these changes occurred with 16 mg/kg dose and at 7 days after treatment, but not at 1 or 3 days. In addition, Tl+-treated animals exhibited general hypokinesis, but no changes were observed in spatial learning. Our findings suggest that a delayed response of the brain to Tl+ may be the result of its residual levels. Also, despite the regional alterations produced by Tl+ in LP and the limited changes in endogenous antioxidants, there is a correlation between the Tl+-induced oxidative damage and the affected behavioral tasks, suggesting that, although still moderate, Tl+ evokes neurotoxic patterns under the experimental conditions tested. PMID- 15833380 TI - Lack of subchronic toxicity of an aqueous extract of Agaricus blazei Murrill in F344 rats. AB - Agaricus blazei Murrill, an edible mushroom, is widely used as a functional food due to its possible medicinal effects. Aqueous extracts are also used as food additive to provide an agreeable bitter taste. As a part of its safety assessment, the present 90-day subchronic toxicity study was performed in F344 rats. To establish a no-observed-adverse-effect level (NOAEL), rats were fed powder diet containing A. blazei Murrill aqueous extract at dose levels of 0 (basal diet), 0.63, 1.25, 2.5 and 5% (maximum) for 90 days. During the experiment, there were no remarkable changes in general appearance and no deaths occurred in any experimental group. Although serum blood urea nitrogen was slightly but significantly increased in males of the 2.5 and 5% groups, no related histopathological changes were observed in the kidney, and serum creatinine levels were rather reduced, suggesting the increase of blood urea nitrogen to be of little toxicological significance. Hematology, organ weight measurement and histopathological observation revealed no test compound-related toxicological changes. In conclusion, A. blazei Murrill extract even at 5% in the diet (2654 mg/kgb.w./day for male rats and 2965 mg/kgb.w./day for female rats) did not cause remarkable adverse effects in F344 rats. Thus, the NOAEL was concluded to be 5% in the diet. PMID- 15833381 TI - Developmental toxicity of two trimethylbenzene isomers, mesitylene and pseudocumene, in rats following inhalation exposure. AB - The developmental toxicity of two trimethylbenzene isomers, mesitylene (1,3,5 trimethylbenzene) and pseudocumene (1,2,4-trimethylbenzene) was studied in Sprague-Dawley rats following inhalation exposure. Pregnant rats were exposed whole body to vapours of mesitylene (0, 100, 300, 600, and 1200 ppm) or pseudocumene (0, 100, 300, 600, and 900 ppm), 6h/day, on gestational days (GD) 6 through 20. Significant decrease in maternal body weight gain and food consumption was observed at concentrations of 300 ppm mesitylene, 600 ppm pseudocumene, or greater. Fetal toxicity, expressed as significant reduction in fetal body weight, occurred at 600 and 1200 ppm mesitylene, and at 600 and 900 ppm pseudocumene. There was no evidence of embryolethal or teratogenic effects following inhalation exposure to either of these chemicals. In summary, the no observed-adverse-effect-level (NOAEL) for maternal toxicity was 100 ppm for mesitylene and 300 ppm for pseudocumene, and the NOAEL for developmental toxicity was 300 ppm for mesitylene and pseudocumene. PMID- 15833382 TI - Lack of carcinogenicity of silicone resin (KS66) in F344 rats. AB - The carcinogenic potential of silicone resin (KS66), used as an antifoaming food additive, was examined in both sexes of F344 rats. Groups of 50 female and 50 male animals were given diet containing KS66 at doses of 0%, 1.25% and 5.0%. No treatment related effects were noted regarding survival rate, general condition, body weight, food consumption, hematology and organ weight data. Detailed histopathological examination revealed no treatment-related increase in the incidences of any non-neoplastic or neoplastic lesions. The results demonstrate that KS66 is not carcinogenic in F344 rats of either sex. PMID- 15833383 TI - Extracts from dulse (Palmaria palmata) are effective antioxidants and inhibitors of cell proliferation in vitro. AB - Previously, we reported that a 1-butanol soluble extract of the edible red alga Palmaria palmata, known as dulse, exhibited hydroxyl and stable free radical scavenging activity as well as inhibition of lipid peroxidation, attributed to the reducing activity and polyphenol content of the dulse extract. In the present study, we evaluated the antioxidant and antiproliferative activities of two grades of dulse harvested from Canadian Maritime locations differing in UV radiation exposure (i.e. west versus east coasts of Grand Manan Island, New Brunswick). The 1-butanol soluble extract from Grade 1 dulse (reduced UV exposure) exhibited lower reducing activity versus Grade 2 dulse (greater UV exposure) reflecting a lower requirement for endogenous antioxidant protection. Grade 1 and 2 dulse extracts both inhibited (p0.03) AAPH-induced lipid peroxidation, but had no effect on AMVN-induced lipid peroxidation, demonstrating the aqueous nature of the antioxidants involved. The Grade 1 and 2 dulse extract inhibition (p<0.05) of HeLa cell proliferation was dose-dependent over 0.5 5.0mg/mL and maximal at 48 and 72h incubation. The antiproliferative effects of the Grade 1 and 2 dulse extracts in the present study likely reflect the bioactivity of the polyphenol content of these extracts. PMID- 15833384 TI - Octyl methoxycinnamate: two generation reproduction toxicity in Wistar rats by dietary administration. AB - Wistar rats continuously received octyl methoxycinnamate (OMC) in the diet through two successive generations at nominal doses of 0, 150, 450 or 1000 mg/kg bw/day. OMC had no adverse effects on estrous cycles, mating behavior, conception, parturition, lactation and weaning, sperm and follicle parameters, macropathology and histopathology of the sexual organs. 1000 mg/kg bw/day reduced parental food consumption and body weight (-14% to -16% in males, -4% to -5% females), increased liver weight, produced hepatic cytoplasmic eosinophilia and erosion/ulceration of glandular stomach mucosa. and led to a slightly decreased implantation rate in the top dose F0 and F1 dams. The high dose F1 and F2 pups had reduced lactation weight gain and organ weights and delayed sexual maturation landmarks. There was no evidence of a selective influence of the test compound on pups' sexual landmarks. The NOAEL (no observed adverse effect level) is 450 mg/kg bw/day for fertility and reproductive performance, for systemic parental and developmental toxicity. PMID- 15833385 TI - The effect of chronic restraint stress and sulfite on visual evoked potentials (VEPs): relation to lipid peroxidation. AB - Stress and sulfite can stimulate numerous pathways leading to an increased production of free radicals which generate a peroxidation cascade producing lipid peroxidation, protein oxidation, DNA damage and cell death, and contribute to the occurrence of pathologic conditions. The purpose of our study was to investigate the effects of stress and sulfite on visual evoked potentials (VEPs) and to examine the relationship between lipid peroxidation and VEP changes. Forty male wistar rats, aged three months were used. They were equally divided into four groups: control (C), the group exposed to restraint stress (R), the group treated with sulfite (S) and the group exposed to stress and treated with sulfite (RS). Chronic restraint stress was applied for 21 days (1h/day) and sodium metabisulfite (520 mg/kg/day) was given by gavage for the same period. All latencies of VEP components were prolonged in the R, S and RS groups with respect to the C group. Brain and retina TBARS levels were found to be increased in those groups compared to the C group. Our results strongly suggest that the latency prolongations of all VEP components may have resulted from the enhancement of lipid peroxidation. Correlation analysis between brain and retina TBARS levels and VEP latencies also support this conclusion. Our data in regard to latency prolongations of all VEP components in the stress group exposed to sulfite, reflected an additive effect of sulfite toxicity on VEP components. Therefore it could be suggested that chronic exposure to stress and sulfite together is associated with a considerable health risk. PMID- 15833386 TI - An approach to investigating the importance of high potency polycyclic aromatic hydrocarbons (PAHs) in the induction of lung cancer by air pollution. AB - Evidence suggests that people living in urban areas have an increased risk of lung cancer due to higher levels of air pollution in these areas. Benzo[a]pyrene (B[a]P) is currently used as the main indicator of carcinogenic polycyclic aromatic hydrocarbons (PAHs) in air pollution, but there is concern that B[a]P may not be the ideal surrogate of choice for PAH mixtures since higher potency PAHs have recently been identified which could potentially contribute more and variably to the overall carcinogenicity. Dibenzo[a,h]anthracene (DBA) and dibenzo[a,l]pyrene (DB[a,l]P) are estimated to have carcinogenic potencies 10 or more times greater than B[a]P but data on their presence and formation in the environment are limited. Several occupational and environmental PAH biomonitoring studies are reviewed here, with particular focus on the specific exposure groups, study design, sample tissue, in particular the use of nasal tissues, and biomarkers used in each study. Consideration of these data is then used to propose a novel biomonitoring approach to evaluate exposure, uptake and the role of high potency PAHs in air pollution-related lung cancer. This is based upon an occupational study examining specific DNA adducts for DBA and DB[a,l]P in nasal cells to evaluate the extent to which these high potency PAHs might contribute to the increased risk of developing lung cancer from air pollution. PMID- 15833387 TI - Liriodenine inhibits the proliferation of human hepatoma cell lines by blocking cell cycle progression and nitric oxide-mediated activation of p53 expression. AB - Liriodenine was isolated from the leaves of Michelia compressa. This study was designed to assess cell cycle arrest, the production of nitric oxide (NO) and p53 expression in liriodenine-treated human hepatoma cell lines, including wild-type p53 (Hep G2 and SK-Hep-1). As evidenced by flowcytometric studies, liriodenine induced cell cycle G(1) arrest and inhibited DNA synthesis in Hep G2 and SK-Hep-1 cell lines. The p53, iNOS expression and intracellular NO level were markedly increased in Hep G2 cells after liriodenine treatment. A NO inhibitor, carboxy PTIO inhibited the p53 expression induced by liriodenine. In addition, liriodenine could not induce obvious cytotoxicity in normal human IMR-90 cell line. These results demonstrate that NO production and p53 expression are critical factors in liriodenine-induced growth inhibition in human wild-type p53 hepatoma cells. PMID- 15833388 TI - A 90-day oral toxicity study of beta-carotene derived from Blakeslea trispora, a natural food colorant, in F344 rats. AB - A subchronic oral toxicity study of beta-carotene derived from Blakeslea trispora, a natural food colorant, was performed with groups of 10 male and 10 female F344 rats fed the agent at dietary levels of 0%, 0.2%, 1.0% and 5.0% for 90 days. There were no treatment-related adverse effects with regard to body weight, food and water consumption, urinalysis, ophthalmology, hematology, serum biochemistry, and organ weight data. On clinical observation, red coloring of fur was noted in both sexes of the 1.0% and 5.0% group rats, with red feces observed in all treated group animals, and necropsy revealed all rats of the treated groups to have reddish coloration of the contents of the gastro-intestinal tract, due to the pigmentation and thus lacking toxicological significance. On histopathological examination, sporadic spontaneous lesions known to occur in this strain of rats were the only findings, with no specific relation to the test substance. Thus, the no-observed-adverse-effect-level (NOAEL) was judged to be a dietary level of at least 5.0% (3127 mg/kg body weight/day for males, 3362 mg/kg body weight/day for females) for beta-carotene derived from B. trispora under the present experimental conditions. PMID- 15833389 TI - Effect of roasting on degradation of Aflatoxins in contaminated pistachio nuts. AB - With increasing knowledge and awareness of aflatoxins (AFs) as potent sources of health hazards to both human and animals, a great deal of effort has been made to completely eliminate the toxin or reduce its content in foods. Although prevention is the most effective intervention, heat has been used to inactivate AFs in contaminated foodstuff. Nuts as general and especially pistachio nuts are very sensitive commodity to AFs contamination. In this study effect of roasting on reduction of AFs content in pistachio nuts has been tested in a laboratory setting with aiming to suggest an optimal condition for the roasting. Although all treatment protocols showed some degree of AFs degradation (ranging from 17% to 63%), roasting spiked samples at 120 degrees C for 120 min and 150 degrees C for 30-120 min caused substantial reduction of AFs in samples. Treatment of naturally contaminated whole pistachio kernels at 150 degrees C for 30 min significantly reduced level of AFs contamination in samples. Degradation of AFs was both time and temperature dependent. Roasting at 150 degrees C and 120 min condition degraded more than 95% of AFB1 in pistachio. However, the resulted product was not edible. AFs in form of naturally occurrence were more resistant to degradation with heat. PMID- 15833390 TI - Shortening of primary dentin etching time and its implication on bond strength. AB - OBJECTIVE: The aim of this study was to investigate the influence of shortening the etching time on the bond strength of a conventional and a self-etching primer adhesive system used in primary tooth dentin. METHODS: Flat dentin surfaces were obtained from 24 primary molars, randomly assigned to 4 experimental groups. The adhesive systems Single Bond and Clearfil SE Bond were applied in two groups according to the manufacturers' recommendations. In the other two groups, the adhesives were applied after half-time of acid etching, 7 s for Single Bond and 10 s for Clearfil SE Primer. Resin crowns were built up and after 24 h storage in water at 37 degrees C, the teeth were sectioned to produce beams with cross sectional area of approximately 0.49 mm2. Specimens were tested in tension at 0.5 mm/min until failure. Fractured specimens were analyzed to determine the failure mode. RESULTS: Tensile bond strengths for Single Bond in primary dentin were higher than for Clearfil SE Bond. Shortening of acid etching time improved bond strength only for Single Bond, while no statistically significant difference was observed for Clearfil SE Bond when both etching times were compared. SIGNIFICANCE: No detrimental effect on bond strength was observed when the time of acid etching was shortened in 50%. Shortening the time for a procedure in a small child without compromising the quality of the work is a very important finding for the practicing pediatric dentist. PMID- 15833391 TI - Effect of hydrogen peroxide on intertubular dentine. AB - OBJECTIVES: To evaluate the effect of 30% hydrogen peroxide on the surface changes and nanomechanical properties of intertubular dentine. METHODS: Five freshly extracted human premolars were sectioned and two specimens were obtained from each tooth. Atomic force microscopy (AFM) images of dentine were taken. Baseline hardness and Young's modulus of intertubular dentine were determined using nanoindentation testing and recorded. The specimens were then treated with either 0.5 ml of Hank's balanced salt solution (control group) or 30% hydrogen peroxide at 24 degrees C for 24 h. AFM imaging and determination of hardness and Young's modulus of intertubular dentine were then repeated. The images were compared and statistical analyses of hardness and Young's modulus were carried out using paired sample t-tests. RESULTS: The AFM images showed recession of the intertubular dentine surface post-bleaching. For the control group, there was no significant change in hardness (p=0.124) and Young's modulus (p=0.438) of intertubular dentine. However, a significant decrease in hardness (p=0.002) and Young's modulus (p=0.001) of intertubular dentine was observed for the experimental group. CONCLUSIONS: Exposure to 30% hydrogen peroxide for 24 h caused surface changes to intertubular dentine and significantly decreased the hardness and Young's modulus of intertubular dentine. PMID- 15833392 TI - Effects of solvents on the early stage stiffening rate of demineralized dentin matrix. AB - OBJECTIVES: To monitor the stiffening rate of demineralized dentin matrix at the early stages after exposure to different neat solvents. METHODS: Dentin beams approximately 0.8x0.7x8.0 mm were obtained from human third molars. After covering their ends with resin composite, the middle exposed length of 4.0mm (gauge-length) was demineralized in 0.5 M EDTA (pH 7.0) for 7 days. The specimens were gripped by a testing machine, pre-loaded to 10 g and cyclically stressed in tension to 5% strain, for 30 repeated cycles (total 20 min) at 0.6 mm/min while immersed in water (control). Then, water was replaced by either 100% acetone, methanol, ethanol, propanol, HEMA or air and the specimens subjected to the same cyclic protocol. The maximum apparent modulus of elasticity (E(Max)) was calculated for every cycle, plotted as a function of time and subjected to regression analysis. Stiffening rate was calculated as changes in E (min). Regression analysis examined the relationship between E and time for each solvent. Data were analyzed by one-way ANOVA and Student-Newman-Keuls test at alpha=0.05. RESULTS: Regression analysis showed that E increased significantly with time in all water-free solvents (R2=0.8-0.99). Stiffening rate was higher for acetone (0.9 MPa/min) and ethanol (0.8 MPa/min), intermediate for air (0.7 MPa/min), methanol (0.6 MPa/min) and propanol (0.5 MPa/min), lower for HEMA (0.2 MPa/min) and practically none for water (0.07 MPa/min) with p<0.05. CONCLUSIONS: The solvent-induced stiffening rate of demineralized dentin matrix is both time and solvent-dependent. The ability of solvents to promptly stiffen the demineralized dentin matrix may be important in maintaining the resin-infiltrated matrix expanded during the solvent evaporation stage of resin bonding. PMID- 15833393 TI - Oral health, sociodemographic factors, dietary and oral hygiene practices in Jordanian children. AB - OBJECTIVES: To investigate the association between oral health, in terms of dental caries and gingivitis, and sociodemographic factors, dental plaque, oral hygiene behaviours, infant feeding and dietary practices in 4-5-year-old Jordanian children. METHODS: Two-stage random sampling procedure was used to select children enrolled in kindergartens in Amman, Jordan. Clinical examinations were carried out by one examiner. Mothers completed questionnaires relating to sociodemographic factors, oral hygiene behaviours, infant feeding and dietary practices of the children. RESULTS: About 67% of the children had caries, 31% had dmft greater than 4 and 66% had gingivitis. Of the children who had been wholly breast-fed, 86% had been fed on demand. Mothers of children attending kindergartens with the lowest tuition fees were more likely to have breast-fed their children on demand and for more than 18 months (prolonged breast feeding) compared to those attending kindergartens with higher fees. Savory snacks were consumed by 82%, confectionery by 76% and teas with sugar by 42% of the children. Multivariate analysis showed age, dental plaque, sleeping beside the mother, use of comforters and selected dietary habits to have an independent effect on caries prevalence. Dental plaque and prolonged breast feeding exerted an independent effect on caries severity and gingivitis. The strongest association with gingivitis was dental plaque. CONCLUSIONS: In Jordan as elsewhere health promotion strategies need to be targeted to mothers from less advantaged backgrounds. Messages about infant breast feeding should emphasise that the method is beneficial to the oral health of the children if appropriately used. PMID- 15833394 TI - Influence of food-simulating solutions and surface finish on susceptibility to staining of aesthetic restorative materials. AB - OBJECTIVES: To determine the degree of surface staining of resin-based composites (RBCs) and glass-ionomer cements (GICs) after immersion in various stains and food-simulating solutions (FSS). METHODS: Six tooth-coloured restorative materials were used: a light-cured microfilled RBC (Durafil, Kulzer), a light cured microglass RBC (Charisma, Kulzer), a polyacid-modified RBC (F2000, 3M/ESPE), a conventional GIC (Fuji IX, GC) and two resin-modified GICs (Fuji II LC, GC; Photac Fil, 3M/ESPE). Disk-shaped specimens were prepared and tested with either a matrix finish or polished using wet silicon carbide papers up to 2000 grit. All specimens were immersed in 37 degrees C distilled water for 1 week, followed by three different FSS (water, 10% ethanol, Crodamol GTCC) and five stains (red wine, coffee, tea, soy sauce and cola) for a further 2 weeks. Three specimens of each material for each stain were tested. Colour coefficients (CIE L* a* b*) were measured by a spectrophotometer after each treatment. The change in colour (DeltaEn) was calculated using the formula: DeltaEn=[(DeltaLn+(Deltaa(n))2+(Deltab(n))2]1/2. RESULTS: Distilled water caused no perceptible colour change as tested by ANOVA and Tukey's tests. The effect of surface finish on staining was not statistically significant (P>0.05). There was no strong interaction between FSS and stains or between FSS and materials. There was a strong interaction between surface and material, and stain and material (P<0.001). CONCLUSIONS: All materials were susceptible to staining by all stains especially coffee, red wine and tea; Fuji IX showed the least susceptibility and F2000 the greatest. PMID- 15833395 TI - Risk analysis of failure of root canal treatment for teeth with inaccessible apical constriction. AB - OBJECTIVE: The risk of failure of root canal therapy was assessed in teeth with inaccessible apical constriction and factors influencing treatment outcome in these teeth were analyzed. METHODS: From the same 57 patients, one tooth with inaccessible apical constriction (n=57) and one tooth with adequate accessibility (n=57) were included in this retrospective investigation. Cox regression analysis was used to analyze factors that may have influenced the outcome one or more years after obturation. These factors included inaccessibility, systemic disease, canal curvature, preoperative periradicular lesion, widening of periodontal ligament space, patient gender and age, size of master apical file, preoperative pulp vitality, years of operator experience and obturation length. RESULTS: Inaccessible apical constriction was a significant factor related to outcome (odds ratio, 5.301). Preoperative presence of periradicular lesion significantly influenced the outcome in cases with inaccessibility (odds ratio, 4.448). CONCLUSION: Inaccessibility increases the risk of root canal therapy failure particularly in teeth with preoperative periradicular lesion. PMID- 15833396 TI - The in-vitro clinical failure of all-ceramic crowns and the connector area of fixed partial dentures: the influence of interfacial surface roughness. AB - OBJECTIVES: To assess the effect of interfacial surface roughness on the flexure strength and fracture mode and origin utilizing an in-vitro assessment of the clinical failure conditions expected for all-ceramic crowns and the connector area of fixed partial dentures (FPDs) using bilayered ceramic specimens tested in bi-axial flexure. METHODS: Sets of 20 bilayered composite discs, with core:dentine thickness ratio of 2:1 and interfacial surface roughnesses determined by alumina abrasion with different alumina particle sizes, were tested in bi-axial flexure with both the reinforcing core and veneering dentine loaded in tension. Mean flexure strengths, standard deviations and associated Weibull Moduli (m) were determined. Optical microscopy was employed for identification of the fracture mode and origin for the failure all-ceramic crowns and the connector area of FPDs. RESULTS: The interfacial surface roughness influenced the bi-axial flexure strength and reliability of the flexure strength data when both the reinforcing core and veneering dentine porcelain were tested in tension. The number of fracture fragments, frequency of occurrence of specimen delaminations, Hertzian cone formations and sub-critical radial cracking in the bilayered dental ceramic composite disc-shaped specimens was also dependent on the interfacial surface roughness and the surface loaded in tension. CONCLUSIONS: The fracture resistance, failure mode and failure origin in bilayered ceramics tested to represent the clinical failure mode of all-ceramic crowns and FPDs are dependent upon the interfacial surface roughness and the modulus of the material in tension. PMID- 15833397 TI - Clinical evaluation of the stain removing ability of a whitening dentifrice and stain controlling system. AB - OBJECTIVES: This study aimed to evaluate the stain removing/controlling properties of a whitening dentifrice and a stain controlling system, in a parallel group, multiple use study. METHODS: Subjects (63), were stratified according to baseline stain scores and randomly allocated to a group, A (standard dentifrice), B (whitening/stain removing dentifrice) or C (whitening/stain removing dentifrice, tooth polish, tooth polisher and whitening/stain removing mouthwash). At each attendance, extrinsic stain was scored blind by a trained and calibrated examiner using the Shaw and Murray Stain Index, under constant lighting conditions. In addition, whiteness was measured using a colorimeter. RESULTS: The toothpaste regimens were well balanced at baseline (p=0.811). At 4 weeks, Regimens B and C had significantly less stain than A (p<0.05). Lower stain values were maintained/reduced for regimens B and C by week 6, with changes from baseline significantly greater than for Regimen A. When lightness was assessed by colorimeter, Regimen C was also found to be significantly whiter than Regimen A (p<0.05). CONCLUSIONS: It was concluded that both whitening regimens had a significant effect on reducing tooth staining and improving whiteness over the 6 week time period. PMID- 15833398 TI - Micro-shear bond strength and morphological analysis of a self-etching primer adhesive system to fluorosed enamel. AB - PURPOSE: The aim of this study was to evaluate micro-shear bond strength and morphological analysis of a self-etching primer adhesive system to fluorosed enamel. MATERIALS AND METHODS: Extracted human molars were classified according to the severity of fluorosis using Thylstrup and Fejerskov index into four groups (TFI: 0, normal; 1-3, mild fluorosis; 3-6, moderate fluorosis; 6-7, severe fluorosis) and divided into following two sub-groups. For the first sub-group, a self-etching primer adhesive system was applied to the ground enamel surfaces and the other sub-group was conditioned with 37% phosphoric acid for 30s prior to application of the same adhesive system. Teeth were then restored with a resin composite, stored for 24h in water and micro-shear bond strengths were measured. After shear testing, the fracture modes were observed under a laser scanning microscope. Morphological study of etching patterns and adhesive interface was done under a scanning electron microscope (SEM). The data were analyzed using two way ANOVA and Sheffe test (p=0.05). RESULTS: No statistically significant difference was there between the different degrees of fluorosis in each sub group. However, significant difference in the bond strengths between phosphoric acid etching and self-etching was found in moderate and severe groups. The SEM observations showed at the resin-enamel interface, thick resin tag like extensions penetrated around 3.5 microm into the enamel etched with phosphoric acid, while self-etching primer created, 1 microm lamina like penetration. CONCLUSION: Severity of fluorosis affects the micro-shear bond strength of a self etching bonding system to fluorosed enamel. PMID- 15833399 TI - Chemical analysis and surface morphology of enamel and dentin following 9.6mu CO2 laser irradiation versus high speed drilling. AB - OBJECTIVES: The purpose of the present in vitro study was to determine whether there is a change in the chemical composition and surface morphology of enamel and dentin following 9.6mu CO2 laser irradiation and high-speed drilling. MATERIALS AND METHODS: Ten permanent, non-carious, young premolars, extracted for orthodontic reasons, were selected. The crowns were separated longitudinally into two equal parts at their mesiodistal axis. Two areas on the inner enamel surface of each specimen and two on the dentinal surface were selected. A high-speed drill and 9.6mu CO2 laser irradiation were applied to the selected enamel and dentinal areas. A random area on the unlased enamel and on the unlased dentin of each specimen served as controls. The morphology of the specimens was evaluated using scanning electron microscopy. Calcium, phosphorus and oxygen levels were measured using an energy dispersive spectrometer. RESULTS: Mineral analysis revealed no significant difference in the mineral content of the enamel and dentin after laser irradiation or high speed drilling versus the control. Use of the high-speed drill on enamel and dentin resulted in very clear cavity margins, with characteristic grooves, whereas laser irradiation of enamel and dentin did not produce clear margins and the floor of the cavity displayed an irregular surface. CONCLUSIONS: The 9.6mu CO2 laser appears to be a promising tool in the clinical setting. However, further investigation is indicated to ensure maximum effectiveness. PMID- 15833400 TI - Longevity of ceramic inlays and onlays luted with a solely light-curing composite resin. AB - OBJECTIVES: To determine the longevity of heat-pressed glass ceramic inlays and onlays luted with solely light-curing composite resin. METHODS: The records of patients who had received a ceramic inlay or onlay at the Heidelberg University Department of Conservative Dentistry from 1993 to 2002 were evaluated. The mean observation time and the survival probability of the ceramic inlays and onlays was calculated with the aid of the Kaplan-Meier algorithm. This was done for the total number of all restorations as well as for various subgroups (number of surfaces, tooth type, endodontic condition at the time that the ceramic restoration was incorporated, experience of operator). The log-rank test was used to compare groups and look for significant differences, and p<0.05 was set to be statistically significant. In addition, 95% confidence intervals of the survival probability values were computed. RESULTS: At the time of the last observation, 783 (96.7%) of these restorations were still in place. The mean observation period for all ceramic restorations was 17.3 months (SD 20.2), with an observation interval of between 0 and 116 months. The last loss of a ceramic restoration was observed 45 months after it had been placed, so that the survival probability of all ceramic restorations amounted to p=0.90 (95% confidence interval 0.86-0.94) from this time onward. Factors such as endodontic condition of tooth, type of tooth, position of tooth, extent of restoration, experience of operator or gender of patient had no significant influence on the survival probability of the ceramic restorations. CONCLUSIONS: Heat-pressed glass ceramic inlays and onlays can be used successfully in routine clinical therapy. In addition, this type of inlays and onlays can be placed successfully with solely light-curing composite resin. PMID- 15833402 TI - Multidisciplinary approach to lung cancer. PMID- 15833401 TI - Secondary caries-like lesions at fissure sealings with Xeno III and Delton--an in vitro study. AB - OBJECTIVES: The aim of the study was to establish a bacterial based in vitro caries model to test the caries preventive effect of fissure sealants and to investigate whether a self-etching adhesive (Xeno III) used as fissure sealant is equally effective in preventing secondary caries compared to a conventional sealant (Delton) when applied on saliva contaminated and non-contaminated fissures. METHODS: Forty caries-free upper premolars were randomly divided into two groups. Group A was sealed with a conventional resin-based fissure sealant (Delton) after phosphoric acid etching. Group B was sealed with a self-etching adhesive (Xeno III) without prior etching. To simulate the clinical situation of saliva contamination, the distal part of each tooth was slightly moistened with saliva before sealant application. After thermo-cycling, the specimens were exposed to a cariogenic Streptococcus mutans biofilm in an artificial mouth model. Depths of demineralization were measured on serial sections by confocal laser scanning microscopy. If a demineralization was detected underneath the whole fissure sealing the sample was rated as 'total failure'. RESULTS: Significantly higher numbers of total failures could be found in the Xeno III group compared to the Delton group in areas without saliva contamination. On contaminated areas, no significant difference could be found. There was no significant difference regarding the depth of the primary carious lesions between groups. CONCLUSION: The study supports our recent findings that our caries model is capable to produce secondary caries-like lesions at fissure sealings. Based on the present in vitro data, it cannot be recommended to use the self-etching adhesive Xeno III solely as a fissure sealant. PMID- 15833403 TI - Lung cancer screening. AB - It has been demonstrated that 85% of all lung cancers are associated with previous or current smoking. A group of patients has been identified that is at a higher risk for lung cancer, stroke, and heart disease. This article reviews previous reports, current consensus statements, problems that are associated with current screening programs, and hopes for the future. PMID- 15833404 TI - Clinical evaluation and staging of patients who have lung cancer. AB - This article updates the performance characteristics of the most commonly used diagnostic and staging modalities in the evaluation of a patient who has lung cancer. Recent data with newer modalities, like positron emission tomography, are reviewed, and their role in this evaluation is discussed. PMID- 15833405 TI - Nonsurgical therapy for stages I and II non-small cell lung cancer. AB - For patients who have stages I and II non-small cell lung cancer (NSCLC) and who are unable or unwilling to undergo surgical resection, nonsurgical treatment modalities have been used with curative intent. Conventionally fractionated radiotherapy has been the mainstay of nonsurgical therapy; however, advances in technology and the clinical application of radiobiologic principles have allowed more accurately targeted treatment that delivers higher effective doses to the tumor, while respecting the tolerance of surrounding normal tissues. This article discusses nonsurgical approaches to the treatment of early-stage NSCLC, including several promising techniques, such as radiation dose escalation, altered radiation fractionation, stereotactic radiotherapy, and radiofrequency ablation. PMID- 15833406 TI - Adjuvant chemotherapy of lung cancer: methodologic issues and therapeutic advances. AB - This article discusses methodologic issues relevant to the interpretation of recent studies focusing on adjuvant chemotherapy of lung cancer. It also attempts to conclude where we currently stand in this field. PMID- 15833407 TI - Postoperative radiation therapy for patients who have resected non-small cell lung cancer. AB - The role of adjuvant postoperative therapy for patients who have resected non small cell lung cancer (NSCLC) has been controversial. Recent large trials and meta-analysis support a modest, but significant, survival benefit for postoperative adjuvant chemotherapy. Postoperative radiation therapy has little potential benefit in patients who have N(0) or N(1) disease and should not be used routinely in these patients. There are suggestions of benefit for patients who have N(2) disease, but adequately powered trials have not been conducted to demonstrate a possible small, but clinically worthwhile, survival benefit in this subgroup. Central nervous system relapse has emerged as an increasing problem for patients who have NSCLC; trials of prophylactic cranial irradiation are underway in several cooperative trial groups. PMID- 15833408 TI - The role of surgery in stage IIIA non-small cell lung cancer. AB - For most patients who have stage III non-small cell lung cancer (NSCLC), a combination of chemotherapy and radiation represents the current treatment of choice. Recent developments include the use of adjuvant chemotherapy after up front surgery in subgroups of patients who have stage III disease, as well as innovative ways to deliver concurrent chemoradiotherapy or combinations of chemotherapy with higher-dose conformal radiotherapy techniques. This article focuses on patients who have stage IIIA NSCLC and reviews the different possibilities for their treatment. Special emphasis is given to the inclusion of surgery into the different approaches to this disease stage. The current literature on this topic is reviewed, and the different aspects of surgical treatment in the management of stage IIIA NSCLC are discussed. PMID- 15833409 TI - Combined chemoradiotherapy in small cell lung cancer. AB - Combined chemoradiotherapy is the established standard of care for limited stage small cell lung cancer; it provides cure in 15% to 25% of patients. Early concurrent therapy imparts a 5% long-term survival benefit compared with sequential therapy. Hyperfractionated delivery of radiotherapy may provide a small incremental benefit when compared with standard fractionation. Radiotherapy dose escalation and reduced radiotherapy volumes are feasible; however, survival benefit has not been confirmed. Cisplatin and etoposide remain the preferred chemotherapy agents. New chemotherapeutic agents and novel treatment approaches are under intense investigation. PMID- 15833410 TI - Novel therapies in lung cancer. AB - This article reviews novel therapies that are under development for non-small cell lung cancer and small cell lung cancer. PMID- 15833411 TI - Supportive care in lung cancer. AB - Lung cancer often is associated with significant morbidity, which has a detrimental effect on quality of life. Supportive care plays a central role in the multimodal treatment of lung cancer. Palliation of symptoms often improves quality of life and compliance with therapy. New developments in supportive care, reviewed here, include management of symptoms of the disease, such as respiratory problems, pain, and cachexia, as well as effects of treatment, including chemotherapy-induced nausea and vomiting, neutropenia, anemia, and mucositis. In the past few years, significant advances have been made in this field; however, palliation of the symptoms of lung cancer remains an area of active investigation. PMID- 15833412 TI - Quality of life in lung cancer: the validity and cross-cultural applicability of the Functional Assessment Of Cancer Therapy-Lung scale. AB - This article discusses quality of life in lung cancer: the validity and cross cultural applicability of the Functional Assessment of Cancer Therapy-Lung scale. PMID- 15833415 TI - The distal metatarsal osteotomy for the treatment of hallux valgus. AB - This article discusses various forms of distal metatarsal osteotomy for the treatment of hallux valgus. The techniques for the various osteotomies have evolved over the years to allow the surgeon to match a procedure and its modifications to the individual patient's deformity, thus optimizing outcomes. Fixation techniques continue to evolve, and meticulous surgical technique to prevent complications remains a must. Regardless of the osteotomy used, the authors believe that adherence to the techniques laid out in current literature will provide gratifying results for the surgeon and the patient. PMID- 15833416 TI - Midshaft first-ray osteotomies for hallux valgus. AB - In this article, the authors describe the indications, contraindications, surgical technique and its fixation considerations, postoperative care, results, and potential complications of the offset V and the traditional Z osteotomies. Because of its versatility, inherent stability, minimal first metatarsal shortening, good intermetatarsal reduction, and ease of rigid internal fixation, shaft osteotomies are gaining popularity as procedures for the correction of hallux valgus. Strong and stable internal fixation allows for earlier functional recovery and primary bone healing. Shaft osteotomies can provide predictable and rewarding results for the patient and surgeon for the correction of hallux valgus. PMID- 15833417 TI - Central metatarsal head-neck osteotomies: indications and operative techniques. AB - The author presents the history and indications of various central metatarsal head-neck osteotomies with special emphasis on the radiographic assessment, vascular supply, and regional anatomy. The surgical techniques described include (1) minimal incision osteotomy; (2) Weil metatarsal osteotomy and various modifications; and (3) a novel "telescoping" osteotomy. Ancillary soft-tissue and osseous digital procedures as well as the prevention and management of potential complications are presented in detail. PMID- 15833418 TI - The tailor's bunionette deformity: a field guide to surgical correction. AB - The author presents a detailed review of the Tailor's bunionette deformity of the fifth metatarsal with special emphasis on radiographic analysis and surgical correction. The surgical techniques discussed include (1) partial metatarsal head ostectomy; (2) metatarsal head resection; (3) minimal incision osteotomy; (4) osteotomies about the metatarsal head-neck, shaft, and base; and (5) ancillary soft tissue procedures. Techniques employed to prevent and correct potential complications are discussed in detail for each osteotomy. PMID- 15833419 TI - Midfoot osteotomies for the cavus foot. AB - Midfoot osteotomies have long been used for a wide variety of congenital and acquired deformities. Severe pes cavus often necessitates some form of surgical correction, and no single procedure can be used exclusively. Midfoot osteotomies may be combined with adjunctive procedures to form an appropriate strategy for the treatment of these deformities. The authors present an overview of pes cavus, including evaluation and classification of the deformity, as well as associated conditions. Several soft tissue and osseous procedures for correction of the cavus foot are discussed. Special attention is directed at the use of various midfoot osteotomies and their ability to enhance such correction. PMID- 15833420 TI - The Evans calcaneal osteotomy. AB - The Evans calcaneal osteotomy is currently the premier procedure for lateral column lengthening of the flexible flatfoot deformity. It has withstood the test of time, proving itself an effective procedure for the correction of pediatric flexible flatfoot. Current understanding of the osteotomy has allowed the Evans calcaneal osteotomy to become a useful tool in the correction of the adult flexible flatfoot as well. PMID- 15833421 TI - Posterior calcaneal displacement osteotomy for the adult acquired flatfoot. AB - The posterior calcaneal displacement osteotomy with flexor digitorum longus tendon transfer is an accepted approach to the stage II posterior tibial tendon dysfunction flatfoot. This reconstructive osteotomy provides a viable alternative to isolated hindfoot arthrodesis procedures. Proper patient selection and sound surgical technique ensure favorable postoperative results. Complications, though limited, may include sural neuritis, peroneal tendonitis, undercorrection, and peritalar arthrosis. PMID- 15833422 TI - Tibial osteotomies for lower extremity deformity correction. AB - Lower extremity deformities and reconstruction requires a thorough understanding of the pathology and the underlining etiologies. This article reviews the basic knowledge of identifying the level of pathology and providing reconstruction of tibial deformities with osteotomies and the use of external fixation. It also provides a brief overview of proximal, midshaft, and distal tibial osteotomies and their indications. PMID- 15833423 TI - Long arm decompression osteotomy for hallux limitus. AB - This article presents a new osteotomy for stage I or II hallux limitus. The long arm decompression osteotomy can be used to shorten and plantarflex the first metatarsal. The indications, surgical technique, advantages, and disadvantages are described in detail. PMID- 15833424 TI - A rapid PCR method for genotyping the Large(myd) mouse, a model of glycosylation deficient congenital muscular dystrophy. AB - The myodystrophy (Large(myd)) mouse has a spontaneous loss of function mutation in a putative glycosyltransferase gene (Large). Mutations in the human gene (LARGE) have been described in congenital muscular dystrophy type 1D (MDC1D). Mutations in four other genes that encode known or putative glycosylation enzymes (POMT1, POMGnT1, fukutin and FKRP) are also associated with muscular dystrophy. In all these diseases hypoglycosylation of alpha-dystroglycan, and consequent loss of ligand binding, is a common pathomechanism. Currently, the Large(myd) mouse is the principal animal model for studying the underlying molecular mechanisms of this group of disorders. Over-expression of LARGE in cells from patients with mutations in POMT1 or POMGnT1 results in hyperglycosylation of alpha-dystroglycan and restoration of laminin binding. Thus, LARGE is a potential therapeutic target. Here, we define the intronic deletion breakpoints of the Large(myd) mutation and describe a simple, PCR-based diagnostic assay, facilitating the study of this important animal model. PMID- 15833425 TI - Proteolysis of beta-dystroglycan in muscular diseases. AB - Alpha-dystroglycan is a cell surface peripheral membrane protein which binds to the extracellular matrix (ECM), while beta-dystroglycan is a type I integral membrane protein which anchors alpha-dystroglycan to the cell membrane via the N terminal extracellular domain. The complex composed of alpha-and beta dystroglycan is called the dystroglycan complex. We reported previously a matrix metalloproteinase (MMP) activity that disrupts the dystroglycan complex by cleaving the extracellular domain of beta-dystroglycan. This MMP creates a characteristic 30 kDa fragment of beta-dystroglycan that is detected by the monoclonal antibody 43DAG/8D5 directed against the C-terminus of beta dystroglycan. We also reported that the 30 kDa fragment of beta-dystroglycan was increased in the skeletal and cardiac muscles of cardiomyopathic hamsters, the model animals of sarcoglycanopathy, and that this resulted in the disruption of the link between the ECM and cell membrane via the dystroglycan complex. In this study, we investigated the proteolysis of beta-dystroglycan in the biopsied skeletal muscles of various human muscular diseases, including sarcoglycanopathy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy, Fukuyama congenital muscular dystrophy, Miyoshi myopathy, LGMD2A, facioscapulohumeral muscular dystrophy, myotonic dystrophy and dermatomyositis/polymyositis. We show that the 30 kDa fragment of beta-dystroglycan is increased significantly in sarcoglycanopathy and DMD, but not in the other diseases. We propose that the proteolysis of beta-dystroglycan may contribute to skeletal muscle degeneration by disrupting the link between the ECM and cell membrane in sarcoglycanopathy and DMD. PMID- 15833426 TI - Congenital muscular dystrophy with glycosylation defects of alpha-dystroglycan in Japan. AB - Glycosylation defects of alpha-dystroglycan (alpha-DG) cause various muscular dystrophies. We performed clinical, pathological and genetic analyses of 62 Japanese patients with congenital muscular dystrophy, whose skeletal muscle showed deficiency of glycosylated form of alpha-DG. We found, the first Japanese patient with congenital muscular dystrophy 1C with a novel compound heterozygous mutation in the fukutin-related protein gene. Fukuyama-type congenital muscular dystrophy was genetically confirmed in 54 of 62 patients. Two patients with muscle-eye-brain disease and one Walker-Warburg syndrome were also genetically confirmed. Four patients had no mutation in any known genes associated with glycosylation of alpha-DG. Interestingly, the molecular mass of alpha-DG in the skeletal muscle was similar and was reduced to approximately 90 kDa among these patients, even though the causative gene and the clinico-pathological severity were different. This result suggests that other factors can modify clinical features of the patients with glycosylation defects of alpha-DG. PMID- 15833427 TI - Preservation of in vitro muscle fiber function in dermatomyositis and inclusion body myositis: a single fiber study. AB - Five patients with untreated dermatomyositis, five with inclusion body myositis, and 16 healthy elderly volunteer subjects (controls) underwent open (dermatomyositis and inclusion body myositis) or percutaneous (controls) muscle biopsy. Biopsied muscles included deltoid, biceps and vastus lateralis. Chemically skinned single muscle fibers were activated with Ca(+2); the slack test was performed to determine maximal unloaded shortening velocity (Vo). Parameters measured include single fiber cross sectional area, maximal force, specific force and Vo. 429 Type I and 94 Type IIA fibers were studied. Cross sectional area and maximal force were greater in inclusion body myositis than dermatomyositis or control for Type I and IIA fibers. Specific force of Type I fibers was similar in inclusion body myositis and dermatomyositis but greater than in controls. Vo was greater in Type I, but not IIA, fibers in dermatomyositis compared with inclusion body myositis and controls. The force and velocity generating capacity of single muscle fibers is preserved in patients with dermatomyositis and inclusion body myositis suggesting that dysfunction of the contractile proteins does not contribute to clinical muscle weakness. PMID- 15833428 TI - Serological follow-up in juvenile myasthenia: clinical and acetylcholine receptor antibody status of patients followed for at least 2 years. AB - Thirty cases of juvenile myasthenia with at least 2 years' follow-up and one measurement of acetylcholine receptor antibody titer were evaluated. Symptoms were initially ocular in 15 (50%) cases; only one developed generalized disease and others remained ocular in 2-24 years' follow-up. Antibodies to acetylcholine receptor (AchRAb) were absent in 12/30 (40%) of all patients initially, but became positive in 5/28 (18%) on further testing in 1-5 years. Spontaneous remission was observed in 6 cases (20%) of whom one became seropositive, and late relapses after 5-11 years' remission were observed in 3 patients with ocular myasthenia, of whom only one developed AchRAbs. Our results show ocular JM has an exceptional tendency to remain confined to these muscles, and delaying the classification of the disease as AchRAb-negative for a few years and repeating serological tests is advisable in the follow-up. PMID- 15833429 TI - Multifocal motor neuropathy, type 1 diabetes and asymptomatic Hashimoto's thyroiditis: an unusual association. AB - Immune mediated mechanisms play a role in some forms of diabetic neuropathies. We studied a 17-year-old man who developed asymmetric weakness and atrophy in both upper arms soon after the diagnosis of type 1 diabetes. Nerve conduction studies demonstrated multifocal motor conduction blocks, and serological investigations revealed subclinical Hashimoto's thyroiditis. Therapy with intravenous immunoglobulin and cyclophosphamide led to clinical recovery. This is the first observation of an association between type 1 diabetes, multifocal motor neuropathy and Hashimoto's thyroiditis. PMID- 15833430 TI - Allelic heterogeneity of GNE gene mutation in two Tunisian families with autosomal recessive inclusion body myopathy. AB - Autosomal recessive hereditary inclusion body myopathy (AR-HIBM), with sparing of the quadriceps, is characterized by adult-onset, with weakness and atrophy of distal lower limb muscles, and typical histopathological findings in muscle biopsy. AR hIBM is associated with mutations in the UDP-N-acetylglucosamine 2 epimerase/N-acetylmannosamine kinase (GNE) gene on chromosome 9p12-13 . We report two unrelated Tunisian families with clinical and pathological features of AR HIBM. One distinct homozygous GNE missense mutation, M712T, previously reported in Middle Eastern Jewish patients, and a newly identified one, L379H, were found in one patient from each family. We conclude that AR HIBM in Tunisia shows an allelic genetic heterogeneity. PMID- 15833431 TI - New phenotypic diversity associated with the mitochondrial tRNA(SerUCN) gene mutation. AB - We performed detailed clinical, histopathological, biochemical, in vitro translation and molecular genetic analysis in patients from two unrelated families harbouring the tRNA(SerUCN) 7472C-insertion mutation. Proband 1 developed a progressive neurodegenerative phenotype characterised by myoclonus, epilepsy, cerebellar ataxia and progressive hearing loss. Proband 2 had a comparatively benign phenotype characterised by isolated myopathy with exercise intolerance. Both patients had the 7472C-insertion mutation in identical proportions and they exhibited a similar muscle biochemical and histopathological phenotype. However, proband 2 also had a previously unreported homoplasmic A to C transition at nucleotide position 7472 in the tRNA(SerUCN) gene. This change lengthens further the homopolymeric C run already expanded by the 7472C insertion. These data extend the phenotypic range associated with the 7472C insertion to include isolated skeletal myopathy, as well as a MERRF-like phenotype. PMID- 15833432 TI - Dilated cardiomyopathy may be an early sign of the C826A Fukutin-related protein mutation. AB - Limb-girdle muscular dystrophy LGMD2I is caused by mutations in the fukutin related protein (FKRP) gene. Clinically, LGMD2I exhibits a great phenotypic variability ranging from severe, rapidly progressive weakness and wasting of the limb-girdle muscles to mild disorders. Here, we present three siblings without clinical signs of muscle dystrophy, but with dilated cardiomyopathy. Elevated serum creatine kinase level and partial fatty degeneration of muscles on MRI indicated subclinical involvement of skeletal muscles. The patients were homozygous for the common C826A mutation in the FKRP gene. Although cardiac involvement in patients with clinically typical LGMD2I was previously described, no patient with dilated cardiomyopathy as the only clinical manifestation of the FKRP mutation was reported so far. PMID- 15833433 TI - Properties of slow- and fast-twitch muscle fibres in a mouse model of amyotrophic lateral sclerosis. AB - This investigation was undertaken to determine if there are altered histological, pathological and contractile properties in presymptomatic or endstage diseased muscle fibres from representative slow-twitch and fast-twitch muscles of SOD1 G93A mice in comparison to wildtype mice. In presymptomatic SOD1 G93A mice, there was no detectable peripheral dysfunction, providing evidence that muscle pathology is secondary to motor neuronal dysfunction. At disease endstage however, single muscle fibre contractile analysis demonstrated that fast-twitch muscle fibres and neuromuscular junctions are preferentially affected by amyotrophic lateral sclerosis-induced denervation, being unable to produce the same levels of force when activated by calcium as muscle fibres from their age matched controls. The levels of transgenic SOD1 expression, aggregation state and activity were also examined in these muscles but there no was no preference for muscle fibre type. Hence, there is no simple correlation between SOD1 protein expression/activity, and muscle fibre type vulnerability in SOD1 G93A mice. PMID- 15833434 TI - 123rd ENMC International Workshop: management and therapy in myotonic dystrophy, 6-8 February 2004, Naarden, The Netherlands. PMID- 15833435 TI - Heme peroxidase clothing and inhibition with polyphenolic substances revealed by molecular modeling. AB - Molecular modeling techniques were applied to study oligomeric derivatives of phenols, which are produced during peroxidase-catalyzed oxidation. The interaction of substrates and oligomers with Arthromyces ramosus peroxidase (ARP) was analyzed by docking and molecular dynamics methods. The most possible interaction site of oligomers is the active center of the peroxidase. The affinity of oligomers increases with increasing length of oligomers. However, the complexed oligomers produce non-productive complexes with the peroxidase. Molecular dynamics studies showed that oligomer-peroxidase complexes are stable. It seems likely that strong and stable, but non-productive docking of the oligomers determinates peroxidase inhibition during the reaction by preventing the access of regular substrates to the active center of the enzyme. PMID- 15833436 TI - Survival analysis of microarray expression data by transformation models. AB - Many microarray experiments involve examining the time elapsed prior to the occurrence of a specific event. One purpose of these studies is to relate the gene expressions to the survival times. The Cox proportional hazards model has been the major tool for analyzing such data. The transformation model provides a viable alternative to the classical Cox's model. We investigate the use of transformation models in microarray survival data in this paper. The transformation model, which can be viewed as a generalization of proportional hazards model and the proportional odds model, is more robust than the proportional hazards model, because it is not susceptible to erroneous results for cases when the assumption of proportional hazards is violated. We analyze a gene expression dataset from Beer et al. [Beer, D.G., Kardia, S.L., Huang, C.C., Giordano, T.J., Levin, A.M., Misek, D.E., Lin, L., Chen, G., Gharib, T.G., Thomas, D.G., Lizyness, M.L., Kuick, R., Hayasaka, S., Taylor, J.M., Iannettoni, M.D., Orringer, M.B., Hanash, S., 2002. Gene-expression profiles predict survival of patients with lung adenocarcinoma. Nat. Med. 8 (8), 816-824] and show that the transformation model provides higher prediction precision than the proportional hazards model. PMID- 15833437 TI - Prediction by support vector machines and analysis by Z-score of poly-L-proline type II conformation based on local sequence. AB - In recent years, the poly-L-proline type II (PPII) conformation has gained more and more importance. This structure plays vital roles in many biological processes. But few studies have been made to predict PPII secondary structures computationally. The support vector machine (SVM) represents a new approach to supervised pattern classification and has been successfully applied to a wide range of pattern recognition problems. In this paper, we present a SVM prediction method of PPII conformation based on local sequence. The overall accuracy for both the independent testing set and estimate of jackknife testing reached approximately 70%. Matthew's correlation coefficient (MCC) could reach 0.4. By comparing the results of training and testing datasets with different sequence identities, we suggest that the performance of this method correlates with the sequence identity of dataset. The parameter of SVM kernel function was an important factor to the performance of this method. The propensities of residues located at different positions were also analyzed. By computing Z-scores, we found that P and G were the two most important residues to PPII structure conformation. PMID- 15833438 TI - A macroscopic kinetic model for DNA polymerase elongation and high-fidelity nucleotide selection. AB - The enzymatically catalyzed template-directed extension of ssDNA/primer complex is an important reaction of extraordinary complexity. The DNA polymerase does not merely facilitate the insertion of dNMP, but it also performs rapid screening of substrates to ensure a high degree of fidelity. Several kinetic studies have determined rate constants and equilibrium constants for the elementary steps that make up the overall pathway. The information is used to develop a macroscopic kinetic model, using an approach described by Ninio [Ninio J., 1987. Alternative to the steady-state method: derivation of reaction rates from first-passage times and pathway probabilities. Proc. Natl. Acad. Sci. U.S.A. 84, 663-667]. The principle idea of the Ninio approach is to track a single template/primer complex over time and to identify the expected behavior. The average time to insert a single nucleotide is a weighted sum of several terms, including the actual time to insert a nucleotide plus delays due to polymerase detachment from either the ternary (template-primer-polymerase) or quaternary (+nucleotide) complexes and time delays associated with the identification and ultimate rejection of an incorrect nucleotide from the binding site. The passage times of all events and their probability of occurrence are expressed in terms of the rate constants of the elementary steps of the reaction pathway. The model accounts for variations in the average insertion time with different nucleotides as well as the influence of G + C content of the sequence in the vicinity of the insertion site. Furthermore the model provides estimates of error frequencies. If nucleotide extension is recognized as a competition between successful insertions and time delaying events, it can be described as a binomial process with a probability distribution. The distribution gives the probability to extend a primer/template complex with a certain number of base pairs and in general it maps annealed complexes into extension products. PMID- 15833439 TI - Cofolga: a genetic algorithm for finding the common folding of two RNAs. AB - In order to predict non-coding RNA genes and functions on the basis of genome sequences, accurate secondary structure prediction is useful. Although single sequence folding programs such as mfold have been successful, it is of great importance to develop a novel approach for further improvement of the prediction performance. In the present paper, a secondary structure prediction method based on genetic algorithm, Cofolga, is proposed. The program developed performs folding and alignment of two homologous RNAs simultaneously. Cofolga was tested with a dataset composed of 13 tRNAs, seven 5S rRNAs, five RNase P RNAs, and five SRP RNAs; as a result, it turned out that the average prediction accuracies for the tRNAs, 5S rRNAs, RNase P RNAs, and SRP RNAs obtained by Cofolga with an optimal weight factor and default parameters were 83.6, 81.8, 73.5, and 67.7%, respectively. These results were superior to those obtained by a single-sequence folding based on free-energy minimization in which corresponding average prediction accuracies were 52.4, 47.4, 57.7, and 52.3%, respectively. Cofolga has a post-processing in which a single-sequence folding is performed after fixation of a predicted common structure; this post-processing enables Cofolga to predict a structure that is present in one of two RNAs alone. The executable files of Cofolga (for Windows/Unix/Mac) can be obtained by an e-mail request. PMID- 15833440 TI - Theoretical aspects of 13C metabolic flux analysis with sole quantification of carbon dioxide labeling. AB - The potential of using sole respirometric CO2 labeling measurement for 13C metabolic flux analysis was investigated by metabolic simulations. For this purpose a model was created, considering all CO2 forming and consuming reactions in the central catabolic and anabolic pathways. To facilitate the interpretation of the simulation results, the underlying metabolic network was parameterized by physiologically meaningful flux parameters such as flux partitioning ratios at metabolic branch points and reaction reversibilities. For real case flux scenarios of the industrial amino acid producer Corynebacterium glutamicum and different commercially available (13)C-labeled tracer substrates, observability and output sensitivity towards key flux parameters was investigated. Metabolic net fluxes in the central metabolism, involving, e.g. glycolysis, pentose phosphate pathway, tricarboxylic acid cycle, anaplerotic carboxylation, and glyoxylate pathway were found to be determinable by the respirometric approach using a combination of [1-13C] and [6-13C] glucose in two parallel studies. The reversibilities of bidirectional reactions influence the isotopic labeling of CO2 only to a negligible degree. On one hand, they therefore cannot be determined. On the other hand, their precise values are not required for the quantification of net fluxes. Computer-aided optimal experimental design was carried out to predict the quality of the information from the respirometric tracer experiments and identify suitable tracer substrates. A combination of [1-13C] and [6-13C] glucose in two parallel studies was found to yield a similar quality of information as compared to an approach with mass spectrometric labeling analysis of secreted products. The quality of information can be further increased by additional studies with [1,2-13C2] or [1,6-13C2] glucose. Respirometric tracer studies with sole labeling analysis of CO2 are therefore promising for 13C metabolic flux analysis. PMID- 15833441 TI - Application of residue distribution along the sequence for discriminating outer membrane proteins. AB - Discriminating outer membrane proteins from other folding types of globular and membrane proteins is an important problem both for detecting outer membrane proteins from genomic sequences and for the successful prediction of their secondary and tertiary structures. In this work, we have systematically analyzed the distribution of amino acid residues in the sequences of globular and outer membrane proteins. We observed that the occurrence of two neighboring aliphatic and polar residues is significantly higher in outer membrane proteins than in globular proteins. From the information about the dipeptide composition we have devised a statistical method for discriminating outer membrane proteins from other globular and membrane proteins. Our approach correctly picked up the outer membrane proteins with an accuracy of 95% for the training set of 337 proteins. On the other hand, our method has correctly excluded the globular proteins at an accuracy of 79% in a non-redundant dataset of 674 proteins. Furthermore, the present method is able to correctly exclude alpha-helical membrane proteins up to an accuracy of 87%. These accuracy levels are comparable to other methods in the literature. The influence of protein size and structural class for discrimination is discussed. PMID- 15833442 TI - A deterministic algorithm for constrained enumeration of transmembrane protein folds. AB - A deterministic algorithm for enumeration of transmembrane protein folds is presented. Using a set of sparse pairwise atomic distance constraints (such as those obtained from chemical cross-linking, FRET, or dipolar EPR experiments), the algorithm performs an exhaustive search of secondary structure element packing conformations distributed throughout the entire conformational space. The end result is a set of distinct protein conformations, which can be scored and refined as part of a process designed for computational elucidation of transmembrane protein structures. PMID- 15833443 TI - Modelling of calcium dynamics in brain energy metabolism and Alzheimer's disease. AB - Functional imaging techniques play a major role in the study of brain activation by monitoring the changes in blood flow and energy metabolism. In order to interpret functional neuroimaging data better, the existing mathematical models describing the links that may exist between electrical activity, energy metabolism and hemodynamics in literature are thoroughly analyzed for their advantages and disadvantages in terms of their prediction of available experimental data. Then, these models are combined within a single model that includes membrane ionic currents, glycolysis, mitochondrial activity, exchanges through the blood-brain barrier, as well as brain hemodynamics. Particular attention is paid to the transport and storage of calcium ions in neurons since calcium is not only an important molecule for signalling in neurons, but it is also essential for memory storage. Multiple efforts have underlined the importance of calcium dependent cellular processes in the biochemical characterization of Alzheimer's disease (AD), suggesting that abnormalities in calcium homeostasis might be involved in the pathophysiology of the disease. The ultimate goal of this study is to investigate the hypotheses about the physiological or biochemical changes in health and disease and to correlate them to measurable physiological parameters obtained from functional neuroimaging data as in the time course of blood oxygenation level dependent (BOLD) signal. When calcium dynamics are included in the model, both BOLD signal and metabolite concentration profiles are shown to exhibit temporal behaviour consistent with the experimental data found in literature. In the case of Alzheimer's disease, the effect of halved cerebral blood flow increase results in a negative BOLD signal implying suppressed neural activity. PMID- 15833444 TI - Ribosome builder: a software project to simulate the ribosome. AB - The Ribosome Builder is a software project that provides tools and techniques to create dynamic models of macromolecular systems from the rapidly growing numbers of atomic structural models. It includes a computer program that allows the user to assemble the multiple molecular components within a 3D space and to define the hypothetical interactions of these components with the initial goal of understanding protein translation at an atomic level of detail. The program employs a simplified molecular dynamics forcefield that can simulate the long time-scale events, such as docking of translation factors and mRNA translocation. An embedded scripting language and Application Programming Interface (API) enable the creation of Steered Molecular Dynamics (SMD) simulations through the programmable application of external forces and torques on atoms and bonds. A graphical interface is provided for displaying and interacting with models, recording movies of molecular dynamics movements, and creating annotated 3D simulations of complex macromolecular events. Initial applications of the project include simulation of tetraloop folding, docking of an mRNA on the 30S subunit and a schematic simulation of the translation elongation cycle. The program is an open source project released under the GNU public license. PMID- 15833445 TI - MSAID: multiple sequence alignment based on a measure of information discrepancy. AB - We propose an algorithm of global multiple sequence alignment that is based on a measure of what we call information discrepancy. The algorithm follows a progressive alignment iteration strategy that makes use of what we call a function of degree of disagreement (FDOD). MSAID begins with distance calculation of pairwise sequences, based on FDOD as a numerical scoring measure. In the next step, the resulting distance matrix is used to construct a guide tree via the neighbor-joining method. The tree is then used to produce a multiple alignment. Current alignment is next used to produce a new matrix and a new tree (with FDOD scoring measure again). This iterative process continues until convergence criteria (or a stopping rule) are satisfied. MSAID was tested and compared with other prior methods by using reference alignments from BAliBASE 2.01. For the alignments with no large N/C-terminal extensions or internal insertions MSAID received the top overall average in the tests. Moreover, the results of testing indicate that MSAID performs as well as other alignment methods with an occasional tendency to perform better than these prior techniques. We, therefore, believe that MSAID is a solid and reliable method of choice, which is often (if not always) superior to other global alignment techniques. PMID- 15833446 TI - MAPK pathway contributes to density- and hypoxia-induced expression of the tumor associated carbonic anhydrase IX. AB - Transcription of the CA9 gene coding for a tumor-associated carbonic anhydrase IX (CA IX) isoform is regulated by hypoxia via the hypoxia-inducible factor 1 (HIF 1) and by high cell density via the phosphatidylinositol-3-kinase (PI3K) pathway. We examined the role of the mitogen-activated protein kinase (MAPK) pathway in the control of CA9 gene expression. Inhibition of MAPK signaling by U0126 in HeLa cells led to reduced activity of the PR1-HRE-luc CA9 promoter construct and decreased CA IX protein levels in dense culture as well as in hypoxia. Similar reduction was obtained by expression of a dominant-negative ERK1 mutant and was also observed in U0126-treated HIF-1alpha-deficient Ka13 cells. Simultaneous treatment with the MAPK and PI3K inhibitors U0126 and LY 294002 had stronger effect than individual inhibition of these pathways. Taken together, our results suggest that besides the PI3K pathway, the MAPK cascade is involved in the regulation of CA9 gene expression under both hypoxia and high cell density. PMID- 15833447 TI - Isolation, sequencing, and functional analysis of the TATA-less human ATPase II promoter. AB - Multiple lines of evidence indicate that the P-type Mg(2+)-ATPase, termed ATPase II, could play an important role in apoptosis. With the long-term objective of studying the regulation of this protein during apoptosis, we delineated the exon intron organization of the human ATPase II gene (within chromosome 4). Subsequently, we used RNA ligase-mediated rapid amplification of cDNA ends to identify a major transcription start site at position -143 with respect to the translation start site. Luciferase reporter analysis of a 1.2-kb 5'-flanking sequence (-1222 to +94 with respect to the transcription start site) revealed strong promoter activity in three human cell lines, human oligodendroglioma (HOG), SHSY5Y (hybrid neuroblastoma), and EA.hy926 (endothelial cell line). Serial deletions from the 5' end of this sequence up to nucleotide -291 yielded some decrease in activity only in the EA.hy926 cells. Further deletion to -217 caused a drastic decrease in activity in all three cell lines, but a -148 fragment showed preferential reduction in activity in the EA.hy926 cells. The promoter activity was nearly equal in two sequence variants of the promoter, one of which (designated as Variant 2) contained a 15-bp direct repeat within a GC rich region. Additionally, there were several single base-pair changes from the sequence reported by the human genome project. Despite the presence of enhancer/repressor elements, such as Sp1 and NFkappaB, relatively small differences in promoter activity were observed in the three cell lines. However, it is likely that such sequence elements could cause major regulation of promoter activity in cells subjected to conditions that trigger apoptosis. The ATPase II promoter sequence will provide valuable clues to the regulation and role of the ATPase II protein. PMID- 15833448 TI - Translational effects of differential codon usage among intragenic domains of new genes in Drosophila. AB - Evolved codon usages often pose a technical challenge over the expressing of eukaryotic genes in microbial systems because of changed translational machinery. In the present study, we investigated the translational effects of intragenic differential codon usage on the expression of the new Drosophila gene, jingwei (jgw), a chimera derived from two unrelated parental genes: Ymp and Adh. We found that jgw possesses a strong intragenic differential usage of synonymous codons, i.e. the Adh-derived C-domain has a significantly higher codon bias than that of the Ymp-derived N-domain (P=0.0023 by t-test). Additional evolutionary analysis revealed the heterogeneous distribution of rare codons, implicating its role in gene regulation and protein translation. The in vitro expression of jgw further demonstrated that the heterogeneous distribution of rare codons has played a role in regulating gene expression, particularly, affecting the quality of protein translation. PMID- 15833450 TI - The evolution of hip resurfacing arthroplasty. AB - Metal-on-metal hip resurfacing, a significant recent development in hip arthroplasty, preserves proximal femoral bone stock, optimizes stress transfer to the proximal femur, and offers inherent stability and optimal range of movement. The results of hip resurfacing in the 1970s and 1980s were disappointing, and the procedure was largely abandoned by the mid-1980s. The renaissance of metal-on metal articulations for total hip arthroplasty has enabled the introduction of new hip resurfacings, and many implant manufacturers have introduced such systems. Early results are encouraging, and complications commonly seen in the 1970s and 1980s, such as early implant loosening and femoral neck fracture, are rare. Background research and better understanding of implant failure suggest that current hip resurfacing technology has developed beyond that of an experimental procedure. PMID- 15833451 TI - Influence of the clearance on in-vitro tribology of large diameter metal-on-metal articulations pertaining to resurfacing hip implants. AB - Large-diameter metal-on-metal articulations may provide an opportunity for wear reduction in total hip implants because earlier studies have shown that the formation of a fluid film that completely separates the bearing surfaces is theoretically possible. In such a lubrication mode and under ideal conditions, there is theoretically no amount of wear. Studies have suggested that the two primary parameters controlling the lubrication mode are the diameter and the clearance of the articulation. The goal of the present study was to experimentally investigate the influence of these two parameters on the wear behavior of large-diameter metal-on-metal articulations pertaining to resurfacing hip implants. The results of this in vitro investigation showed that longer running-in periods and higher amounts of running-in wear were associated with larger clearances. PMID- 15833452 TI - Current concepts of metal-on-metal hip resurfacing. AB - The second-generation, metal-on-metal (MOM) bearing for total hip replacements was launched in the 1980s, and resurfacing followed in the mid-1990s. Remaining challenges include long-term bone remodeling of the femoral resurfacing and consideration of adverse MOM wear conditions. Precise understanding of manufacturing variables such as alloy types, bearing diameters, design tolerances, and surface finish is imperative in obtaining clinical consistency and safety in the patient. This review examines femoral fixation, bone remodeling, and wear studies of MOM implants and provides a brief overview of the latest outcome and retrieval data and how these data integrate with the in vitro wear studies. PMID- 15833453 TI - Anatomic considerations for the choice of surgical approach for hip resurfacing arthroplasty. AB - A number of surgical exposures have been advocated over the past 20 years by the pioneers of resurfacing hip arthroplasty and include the anterior, anterolateral, lateral, and posterolateral approaches. Not all of these approaches, however, appear to provide adequate exposure while respecting the local biology that seems to be imperative for the procedure. Based on an anatomic study, the most "bio logical" surgical approach for hip resurfacing arthroplasty appears to be through a lateral or posterolateral approach using a digastric trochanteric osteotomy combined with an anterior hip dislocation. These exposures avoid injury to the medial femoral circumflex artery supplying the femoral head and allow access and treatment to the commonly observed hip pathologies that are frequently located anteriorly. PMID- 15833454 TI - Distribution of vascular foramina around the femoral head and neck junction: relevance for conservative intracapsular procedures of the hip. AB - Precise knowledge of femoral head vascular anatomy is mandatory for the safe execution of intracapsular surgical procedures aimed at preserving the hip joint. No quantitative topographic data exist concerning the exact location and distribution of the entrance of these vessels through nutrient vascular foramina at the femoral head and neck junction. This study shows that only a few significant retinacular vessels may reach the adult femoral head on its anterior aspect and that most vascular foramina are located close to the antero- and posterosuperior regions of the femoral head and neck junction. This larger segment should be carefully protected during any conservative intracapsular procedure of the hip and may be desirable in femoral head resurfacing arthroplasty. PMID- 15833455 TI - Patient selection and surgical technique for surface arthroplasty of the hip. AB - Factors such as patient selection, surgical technique, and durable fixation of the components are critical in surface arthroplasty of the hip. The Surface Arthroplasty Risk Index helps determine the risk of failure, with an index score Zn > Fe > Cu >> Cd > Pb > Se > Mn >> Hg. The results of speciation of arsenic demonstrate that virtually all arsenic was in the arsenobetaine form, which is the less toxic form. The concentrations of several trace elements were generally high in samples from Viana in 2002. Cadmium concentrations were above the legal limit for human consumption in samples from Viana in 2002 and two of these animals also had lead concentrations that exceeded legal limits. Mercury appeared in all samples but levels were within legally defined safe limits. No relationship was detected between trace element concentrations and size or maturity of octopus. PMID- 15833486 TI - Comparative assessment of ambient air quality in two typical Mediterranean coastal cities in Greece. AB - Air quality data (O3, NO2, NO, CO and SO2) of two Greek coastal cities, Patras and Volos, were analyzed and compared to evaluate: (a) the exceedances of air quality EU threshold values, (b) the diurnal patterns of air pollutants and (c) the "weekend effect" on ozone levels. High ozone levels, close to the thresholds for human health and clearly above the threshold for the protection of plants and ecosystems, were observed in Volos. O3 levels in Volos were higher than those in Patras. NOx levels in Patras were significantly higher than the limits for human health and plants' protection. Both, NOx and SO2 levels were higher in Patras than in Volos. The Patras' harbor high traffic seems to drive the diurnal pattern of SO2 in that city. The examination of the rate of ozone accumulation, during the high O3 period (Apr.-Sep.), revealed the occurrence of two phases, a fast and a slow one, with different durations in each city. We suggest that the occurrence of such two phases' patterns should be considered in relevant ozone studies. In both towns, the O3 levels were higher during weekends in comparison to midweek days, although NO levels were lower. Our results support the hypothesis that the weekend O3 effect is due to a combination of VOC sensitivity of the studied areas and the reduced NOx emissions during weekends. Based on the comparison of the weekend effect in the two cities, we suggest the occurrence of a feedback mechanism between peri-urban natural ecosystems (forests) and the polluting anthropogenic ones (cities). PMID- 15833487 TI - Light-induced degradation of metsulfuron-methyl in water. AB - Photodegradation of metsulfuron-methyl, a sulfonylurea herbicide, has been investigated in aqueous solution at different pH and excitation wavelengths. The efficiency of the process has been evaluated through quantum yield determinations. The identification of the photoproducts indicates that the major photochemical pathway is initiated by C-S bond dissociation followed by involvement of water to yield the main final products; the behaviour in water is shown to differ markedly from that in an organic environment. PMID- 15833488 TI - Alkylphenols and alkylphenol ethoxylates contamination of crustaceans and fishes from the Adriatic Sea (Italy). AB - This paper presents the results of an investigation on the occurrence of alkylphenols (APs) and their ethoxylates (APEs) in 8 edible marine species from the Adriatic Sea and tries to estimate the corresponding intake for the Italian population. Two crustaceans, Nephrops norvegicus (Norway lobster) and Squilla mantis (spottail mantis shrimp), plus six fish species, Engraulis enchrascicolus (anchovy), Scomber scombrus (Atlantic mackerel), Merluccius merluccius (European hake), Mullus barbatus (red mullet), Solea vulgaris (common sole) and Lophius piscatorius (angler) were analyzed for their content of nonylphenol (NP), octylphenol (OP) and octylphenol polyethoxylates (OPEs). These compounds were found in all analysed samples. NP was detected at the highest concentrations: 118 399 and 9.5-1431 ng g(-1) fresh weight (fw) respectively in crustaceans and fish. OP was found at respective levels of 2.7-4.7 and 0.3-3.8 ng g(-1) fw in crustaceans and fish, whereas OPE was determined at respective concentrations of 1.2-16.8 and 0.2-21.1 ng g(-1) fw in the same species. These results, together with those from a previous study on 4 edible mollusc, allow to estimate respective daily intakes for NP, OP, and OPE of about 12, 0.1, and 0.1 microg day(-1) for an Italian adult living along the Adriatic Coast. In relation to NP and OP, these intakes are much lower than the doses associated with toxic effects in laboratory animals (9 mg kg(-1) bw for rats). Nevertheless, data of exposure from other sources to these chemicals and others with similar biological characteristics are needed. PMID- 15833489 TI - Lead contamination in tea garden soils and factors affecting its bioavailability. AB - The consumption of heavy metals is detrimental to human health and most countries restrict the concentration of metals such as lead (Pb) in food and beverages. Recent tests have detected high Pb concentrations in certain commercial brands of tea leaves and this finding has raised concerns for both producers and consumers. To investigate what factors may be contributing to the increase in Pb accumulation in the tea leaves we collected tea leaves and soils from tea producing areas and analyzed them for Pb concentration, pH and organic matter content. The result showed the Pb concentration of 47% investigated tea leaves samples was beyond 2 mg kg(-1), the permissible levels given by China. The total Pb concentration in the surface and subsurface soil layers averaged 36.4 and 32.2 mg kg(-1), respectively which fall below of the 60 mg kg(-1) limit provided for organic tea gardens in China. The pH of the tea garden soils was severely acidic with the lowest pH of 3.37. Soils under older tea gardens tended to have a lower pH and a higher Pb bioavailability which was defined as the amount of lead extracted by CaCl2 solution than those under younger tea gardens. We found that the concentration of bioavailable Pb and the percentage of bioavailable Pb (bioavailable Pb relative to total Pb concentration) were positively correlated with soil H+ activity and soil organic matter content, and the organic matter accumulation contribute more effects on Pb bioavailability in these two factors. We conclude that soil acidification and organic matter accumulation could contribute to increasing Pb bioavailability in soil and that these could increase Pb uptake and accumulation in the tea leaves. PMID- 15833490 TI - Simultaneous determination of Cd(II), Cu(II), Pb(II) and Zn(II) by derivative stripping chronopotentiometry in Pittosporum tobira leaves: a measurement of local atmospheric pollution in Messina (Sicily, Italy). AB - The purpose of this study is to investigate the bio-accumulation of Cd(II), Cu(II), Pb(II) and Zn(II) in Pittosporum tobira (Thunb.) Aiton leaves sampled in different zones of Messina, in order to assess the level of atmospheric metal deposition in correlation with the traffic volume. Derivative stripping chronopotentiometry was used as a practical, precise and sensitive technique to determine simultaneously Cd, Cu, Pb and Zn levels in Pittosporum leaves. In the optimised electro-chemical conditions, detection limits lower than 0.05 microg kg(-1) were achieved, whereas the accuracy, expressed as obtained recoveries from certified materials, was in the range 93.5-102.7%. The obtained data provided evidence that Cd and Pb levels significantly decreased from high to low traffic density zones (p < 0.005, ANOVA), whereas Cu and Zn are accumulated by plants particularly from the soil and their contents is not related to the traffic volume. PMID- 15833491 TI - Adsorption and removal of As(V) and As(III) using Zr-loaded lysine diacetic acid chelating resin. AB - An adsorption process for the removal of As(V) and As(III) was evaluated under various conditions using zirconium(IV) loaded chelating resin (Zr-LDA) with lysine-Nalpha,Nalpha diacetic acid functional groups. Arsenate ions strongly adsorbed in the pH range from 2 to 5, while arsenite was adsorbed between pH 7 and 10.5. The sorption mechanism is an additional complexation between arsenate or arsenite and Zr complex of LDA. Adsorption isotherm data could be well interpreted by Langmuir equation for As(V) at pH 4 and As(III) at pH 9 with a binding constant 227.93 and 270.47 dm3 mol(-1) and capacity constant 0.656 and 1.1843 mmol g(-1), respectively. Regeneration of the resin was carried out for As(V) using 1 M NaOH. Six adsorption/desorption cycles were performed without significant decrease in the uptake performance. Column adsorption studies showed that the adsorption of As(V) is more favorable compared to As(III), due to the faster kinetics of As(V) compared to As(III). Influence of the coexisting ions on the adsorption of As(V) and As(III) was studied. The applicability of the method for practical water samples was studied. PMID- 15833492 TI - Heavy metal concentrations in molluscs from the Atlantic coast of southern Spain. AB - Trace metals were determined in the two most abundant species of bivalve molluscs along the Atlantic coast of southern Spain (Donax trunculus and Chamelea gallina) and in the sediments where they live. The results show that the area near the mouth of the Huelva estuary is where the highest metal concentrations are found in sediments and in the two bivalve species. This is not surprising, considering that the Huelva estuary is the mouth of the Tinto and Odiel rivers, which have one of the highest levels of metal pollution of all the rivers of Europe. The two species of bivalves have different amounts of metals in their tissues. The concentrations of Cr, Cu, Pb, Zn, As and Hg in D. trunculus were significantly higher (p < 0.05) than in C. gallina; however, C. gallina contained more Ni and Cd. In both species the most abundant elements were Cu and Zn, while Hg showed the lowest values. There is a significant correlation (p < 0.05) for concentrations of Cu, Pb, Zn and Hg in D. trunculus and C. gallina relative to their concentrations in surface sediments. PMID- 15833493 TI - Statistical characterization of atmospheric PM10 and PM2.5 concentrations at a non-impacted suburban site of Istanbul, Turkey. AB - Inhalable particulate matter (PM10) has been monitored at several stations by Istanbul Municipality. On the other hand, information about fine fraction aerosols (PM2.5) in Istanbul atmosphere was not reported. In this study, 86 daily aerosol samples were collected between July 2002 and July 2003. The PM10 annual arithmetic mean value of 47.1 microg m(-3), was lower than the Turkish air quality standard of 60 microg m(-3). On the other hand, this value was found higher than the annual European Union air quality PM(10) standard of 40 microg m( 3). Furthermore, the annual mean concentration of PM2.5 20.8 microg m(-3) was found higher than The United States EPA standard of 15 microg m(-3). The statistics and relationships of fine, coarse, and inhalable particles were studied. Cyclic behavior of the monthly average concentrations of PM10 and PM2.5 data were investigated. Several frequency distribution functions were used to fit the measured data. According to Chi-squared and Kolmogorov-Smirnov tests, the frequency distributions of PM2.5 and PM10 data were found to fit Log-logistic functions. PMID- 15833494 TI - Di-(2-ethylhexyl)phthalate in sewage sludge and post-treated sludge: quantitative determination by HRGC-MS and mass spectral characterization. AB - DEHP was quantified into sewage sludge, thermally dried sludge and compost samples from different areas of Catalonia (North East Spain) using high resolution gas chromatography (HRGC) coupled to a mass spectrometry (MS). The aim of the work was to determine the evolution of this ubiquitous environmental contaminant along some post-treatment sludge processes, such as composting and thermally drying. DEHP concentration detected in some sludges are above the maximum limit suggested by the EC (100 ppm d.m.) for agricultural use. However, the composting and thermally drying reduce the amount of DEHP in sewage sludge to acceptable level for safe land application. The study was completed with the characterization and long-term observation of DEHP in sewage sludge, composted sludge and thermally dried sludge mixed with two types of soils. In all cases, the percentage of DEHP degraded was higher than 50% after 9 months of incubation. PMID- 15833495 TI - Size distribution of airborne particulate matter and associated heavy metals in the roadside environment. AB - The size distributions of airborne particulate matter (PM) and associated heavy metals Pb, Cd, Ni, Cr, V, Mn, Cu and Fe in different inhalable fractions (< 0.8 microm, 0.8-1.3 microm, 1.3-2.7 microm, 2.7-6.7 microm and > 6.7 microm) were determined at a traffic-orientated urban site in the city of Thessaloniki, northern Greece. The airborne PM displayed a bimodal distribution with most of the mass (52%) contained in the submicron size range (< 0.8 microm) and an additional minor mode (20%) in the coarse size fraction (> 6.7 microm). Characteristic size distributions of heavy metals allowed identification of three main behavioral types: (a) metals whose mass was resided mainly within the accumulation mode (Pb,Cd), (b) those which were distributed between fine, intermediate and coarse modes (Ni,Cu,Mn), and (c) those which were mainly found within particles larger than 2.7 microm in diameter (Fe). The mean mass median aerodynamic diameter (MMAD) of PM was found at 0.85+/-0.71 microm, while the mean MMADs of heavy metals followed the order Pb (0.96 +/- 0.71 microm) < Cd (1.14 +/- 0.82 microm) < V (1.38 +/- 0.63 microm) < Ni (1.45 +/- 0.88 microm) < Cu (2.04 +/ 0.77 microm) < Mn (2.61 +/- 1.23 microm) < Cr (2.91 +/- 1.40 microm) < Fe (3.82 +/- 0.88 microm). The measured distributions are believed to result from a combination of processes including local anthropogenic and natural sources, such as traffic, industrial emissions and resuspension of road dust. PMID- 15833496 TI - Modeling microbial-mediated reduction in batch reactors. AB - The governing equations that depict microbially-mediated reduction of heavy metals in the subsurface include a system of coupled nonlinear partial differential equations (PDE's) that describe physical (transport), chemical (sorption), and microbial (reduction/oxidation) processes. The existence of nonlinear reaction terms makes numerical simulations more challenging; however, with the advent of time-splitting solution algorithms, nonlinear reaction terms can be isolated from the convective-dispersive components of the governing transport equations and then solved as a coupled system of nonlinear ordinary differential equations (ODE's). In this paper, four methods are evaluated for solving coupled systems of nonlinear ODE's that describe microbially-mediated reduction/oxidation processes. The evaluation involves a series of comparisons of transient simulations of electron donor oxidation, electron acceptor reduction, and microbial biomass accumulation. The methods evaluation is initiated with a comparison of simulation results obtained with the four methods to those generated with an analytical model. Next, laboratory observations, of nitrite consumption by Nitrobacter winogradski in batch reactors are used in a comparison of batch system simulations generated using each of the four methods and BIOKEMOD (biogeochemical kinetic/equilibrium reaction model). The evaluation finds one of the four methods, the quasi-steady-state approximation (QSSA), to be among the most accurate and easiest to implement. Final validation of the QSSA is performed simulating experimental results of microbially-mediated chromium reductions in batch cultures. PMID- 15833497 TI - Immunomodulators for treatment of interstitial cystitis. PMID- 15833498 TI - Laparoscopic port site metastases: incidence, risk factors, and potential preventive measures. PMID- 15833499 TI - Peyronie's disease associated with increase in plasminogen activator inhibitor in fibrotic plaque. AB - OBJECTIVES: To investigate whether tissue expression of plasminogen activator inhibitor type 1 (PAI-1) is increased in the fibrotic plaque of human Peyronie's disease (PD). Increased tissue levels of PAI-1, an inhibitor of both fibrinolysis and collagenolysis, have been found in a variety of fibrotic conditions. Recently, it was reported that PAI-1 expression was also increased in the fibrotic plaque of an animal model of PD induced by the injection of fibrin into the tunica albuginea (TA) of the penis. METHODS: Tissue (n = 10/group) and cells (n = 4/group) obtained from the penile TA plaque of patients with PD or from normal TA were subjected to RNA extraction and real-time reverse transcriptase polymerase chain reaction. Tissues were also analyzed by immunohistochemistry (n = 8/group) for the detection of PAI-1 expression at the transcription and protein levels. RESULTS: A significant 3.5-fold to 16-fold increase was found in both PAI 1 mRNA and protein levels in the human PD plaque and the respective fibroblast cultures compared with the normal non-PD TA. CONCLUSIONS: The observed increase in PAI-1 in the human PD plaque agrees with what has been observed in the rat and suggests that PAI-1 may be a key pro-fibrotic factor in the development of human PD. PMID- 15833500 TI - Early hyperbaric oxygen therapy improves outcome for radiation-induced hemorrhagic cystitis. AB - OBJECTIVES: To assess the clinical factors that affect the efficacy of hyperbaric oxygen (HBO2) therapy in treating radiation-induced hemorrhagic cystitis. HBO2 therapy is an effective treatment for radiation-induced hemorrhagic cystitis, with reported response rates ranging from 76% to 100%. METHODS: The data from patients with radiation-induced hemorrhagic cystitis treated at our institution between May 1988 and December 2001 were reviewed retrospectively. All patients received HBO2 therapy at 2.36 atm absolute pressure, with 90 minutes of 100% oxygen breathing per treatment. The outcome was assessed after at least 12 months of follow-up. We evaluated patient demographics, types of pelvic malignancy and radiotherapy, total radiation dose, onset and severity of hematuria, and prior intravesical management. Clinical improvement was defined as the absence of, or reduction in, macroscopic hematuria. RESULTS: A total of 60 patients (55 men and 5 women), mean age 70 years, received an average of 33 HBO2 treatments (range 9 to 63). Of the 60 patients, 48 (80%) had either total or partial resolution of hematuria. When treated within 6 months of hematuria onset, 96% (27 of 28) had complete or partial symptomatic resolution (P = 0.003). All 11 patients with previous clot retention had clinical improvement if treated within 6 months of hematuria onset (P = 0.007). Prior intravesical chemical instillation did not affect the clinical outcome. Patients who had undergone primary, adjuvant, or salvage external beam pelvic radiotherapy showed response rates of 81%, 83%, and 78%, respectively (P = 0.950). CONCLUSIONS: Our results show that delivery of HBO2 therapy within 6 months of hematuria onset is associated with a greater therapeutic response rate. Treatment efficacy was independent of prior intravesical therapy and the timing of radiotherapy. PMID- 15833501 TI - Randomized, double-blind, dose-ranging study of pentosan polysulfate sodium for interstitial cystitis. AB - OBJECTIVES: To compare the current recommended dose of pentosan polysulfate sodium (PPS) with doses two to three times higher. METHODS: We evaluated three dosages (300, 600, and 900 mg) of PPS in a randomized, double-blind, double dummy, parallel-group, multicenter, 32-week study. Adults (n = 380) with a diagnosis of interstitial cystitis (IC) as determined by a positive cystoscopic examination combined with bladder pain and urgency or a history of IC symptoms for at least 6 months were enrolled. Participants completed the Patient's Overall Rating of Symptom Index (PORIS) and the O'Leary-Sant Interstitial Cystitis Symptom Index (ICSI) at baseline (ICSI only) and during follow-up visits at 4, 8, 12, 16, 24, and 32 weeks. RESULTS: Mean ICSI scores improved significantly during the 32 weeks for all dosages (baseline 11.2, 11.9, and 11.9 to endpoint 8.2, 8.1, 8.6 for 300, 600, and 900 mg, respectively; P <0.001) but the response to treatment was not dose dependent (no statistically significant difference in response among the three dosages). At baseline, 3.2%, 62.2%, and 34.6% reported mild, moderate, and severe symptoms, respectively, as assessed by the ICSI. At study end, 27.5%, 56.9%, and 15.7% reported mild, moderate, and severe symptoms, respectively. The PORIS scores improved within 4 weeks with 15.8% to 21.1% of all patients classified as responders (50% or greater improvement on PORIS). At 32 weeks, 49.6%, 49.6%, and 45.2% of all patients were responders at a dose of 300, 600, and 900 mg, respectively. Most adverse events were mild and resolved without intervention. CONCLUSIONS: For all three dosages of PPS, a clinically significant but similar response was demonstrated. The duration of therapy appears to be more important than the dosage. PMID- 15833502 TI - Comparative study of percutaneous access for staghorn calculi. AB - OBJECTIVES: To compare the success and complication rates of staghorn calculi treated through the upper pole calix, lower/middle calices, and multiple access. METHODS: Between January 1995 and December 2000, the records of 119 patients with staghorn calculi who underwent percutaneous nephrolithotomy were retrospectively reviewed. The mean stone burden was 6.9 cm. Of the 119 patients, 16 (13.4%) had upper pole access, 70 (58.8%) lower or middle calix access, and 33 (27.7%) multiple renal access. RESULTS: The stone-free rate was 80.0% for those in the lower/middle, 87.5% for those in the upper pole, and 84.8% for those in the multiple access group. Single access was performed in 72.3% and multiple access tracts in 27.7% of patients. The average operative time was 86.8 minutes in the upper pole, 139.1 minutes in the lower/middle pole, and 134.9 minutes in the multiple access groups. Extracorporeal shock wave lithotripsy was used in 4.2% of patients. The overall complication rate was 28.5% (25% in the upper pole, 21.4% in the lower/middle, and 45.4% in the multiple access group). Thoracic complications (pneumothorax and hydrothorax) occurred in 2 patients (1.7%), but only 1 patient had undergone supracostal access. Bleeding requiring blood transfusion was significantly greater in the multiple access group. The average hospital stay was 3.0 days for all patients. CONCLUSIONS: The success of percutaneous treatment of patients with staghorn calculi is highly related to optimal kidney access. The supracostal and multiple access approaches can be used, with a slight increase in the incidence of acceptable complications. PMID- 15833505 TI - Validation of Spanish version of Pelvic Pain and Urgency/Frequency (PUF) patient symptom scale. AB - OBJECTIVES: To translate the previously described Pelvic Pain and Urgency/Frequency (PUF) questionnaire into Spanish using a back-translation technique and to validate the Spanish version in a group of bilingual women with symptoms consistent with interstitial cystitis or painful bladder syndrome. METHODS: Bilingual women with complaints of urinary urgency/frequency and/or pelvic pain were randomized to complete initially either the Spanish version or the original English version of the PUF questionnaire followed by the questionnaire in the other language. To evaluate retest reliability, subjects completed the Spanish version a second time 1 week later. Demographic information, including age, ethnic origin, and primary language, was obtained. Paired t tests and Wilcoxon signed rank tests for total PUF score, its two domain scores, and each item were used to assess the difference between the Spanish and English versions. Test and retest reliability of the Spanish version was similarly assessed. Agreement between the Spanish and English versions was assessed by weighted kappa statistics and 95% confidence intervals for each item. P values less than 0.05 were considered significant, and kappa values greater than 0.7 were considered to indicate good agreement. RESULTS: No statistically significant difference was found in the mean or median scores (ie, item, symptom, bother, total) between the English and Spanish versions. Good agreement between English and Spanish versions in all eight items was demonstrated by weighted kappa statistics. The Spanish version demonstrated retest reliability among total scores and seven of eight items when administered 1 week later. CONCLUSIONS: The Spanish PUF questionnaire is a valid and reliable instrument for the evaluation and treatment of patients with interstitial cystitis or painful bladder syndrome. PMID- 15833506 TI - Prostate botulinum A toxin injection--an alternative treatment for benign prostatic obstruction in poor surgical candidates. AB - OBJECTIVES: To evaluate, in a prospective study, the effectiveness of prostate injection of botulinum A toxin in patients who were poor surgical candidates. Patients with benign prostatic hyperplasia (BPH) are usually successfully treated with medical treatment or transurethral resection. However, some patients with chronic urinary retention or a large postvoid residual urine volume due to BPH are poor surgical candidates or are patients in whom medical treatment has failed. METHODS: Ten patients with BPH and urinary retention or a large postvoid residual urine volume received 200 U botulinum A toxin injection into the transition zone of the prostate. The clinical results and urodynamic parameters at baseline and after treatment were compared. RESULTS: All patients had an improvement in spontaneous voiding after treatment. Of them, 8 had an excellent result (80%) and 2 had an improved result. Both voiding pressure and postvoid residual volume were significantly decreased after treatment. The total prostate volume was significantly reduced, and the maximal flow rate was significantly increased after treatment. The maximal effects of botulinum A toxin appeared at about 1 week and were maintained at 3 and 6 months after treatment. At 6 to 12 months (mean 9) of follow-up, no patient had had recurrence of urinary retention and the voiding condition in all patients remained at the post-treatment status. No adverse effect was noted. CONCLUSIONS: Prostate injection of botulinum A toxin is an effective alternative treatment with minimal adverse effects for patients with benign prostatic obstruction who are poor surgical candidates or in whom medical treatment has failed. PMID- 15833507 TI - Impact of lymphadenectomy and nodal burden in renal cell carcinoma: retrospective analysis of the National Surveillance, Epidemiology, and End Results database. AB - OBJECTIVES: To analyze the additional impact of retroperitoneal lymphadenectomy on overall and cancer-specific survival (CSS) in patients with primary renal cell carcinoma (RCC) undergoing radical nephrectomy. The benefit of regional lymphadenectomy in patients with primary RCC remains controversial. METHODS: Of 33,016 patients diagnosed with primary RCC between 1983 and 1998, a subset of 4453 underwent radical nephrectomy with or without regional lymphadenectomy. The extent of lymphadenectomy was assessed using the number of nodes examined, and the tumor burden was assessed using the number of positive nodes and the ratio of the number of positive nodes to the total number of nodes examined. Associations between CSS and the number of nodes examined, number of positive nodes, and ratio of the number of positive nodes to the total number of nodes examined were assessed. RESULTS: An inverse correlation was found between the likelihood of CSS and the number of nodes examined, particularly for those with regional disease even after controlling for other factors. A correlation was noted between the number of nodes examined and the number of positive nodes. Significant differences in CSS were observed in node-negative patients with regional disease compared with node-positive patients. An increasing nodal burden was associated with worse CSS. CONCLUSIONS: More extensive lymphadenectomy does not appear to increase further the probability of CSS in patients undergoing radical nephrectomy for RCC. An increased number of positive nodes, as well as an increasing nodal burden, although associated with a lower likelihood of survival, were not independent predictors of RCC-specific mortality. PMID- 15833508 TI - Chromophobe renal cell carcinoma: comprehensive analysis of 104 cases from multicenter European database. AB - OBJECTIVES: To analyze the clinical behavior of chromophobe renal cell carcinoma (CRCC), we retrospectively evaluated the data from six European centers. In 1985, CRCC was identified as a new RCC histologic subtype. Because of its low frequency, only few large CRCC series are available. METHODS: We created a renal cancer database including 3228 patients who underwent surgery between 1986 and 2002 in six European centers. The relevant clinical and pathologic data were extracted from the clinical charts at each institution and collected into a unique database. RESULTS: Of the 3228 patients, 104 (3.2%) affected by CRCC were identified. The mean age at diagnosis was 57.6 years (range 22 to 83). Of the 104 patients, 51 (49%) were men and 53 (51%) were women. The mean tumor size was 6.4 +/- 3.6 cm. An incidental diagnosis accounted for 61.5% of the cases. Radical nephrectomy was performed in 88 patients (85%). After a median follow-up of 38 months (mean 44, range 1 to 153), no local recurrence was observed. The 5-year overall survival rate for CRCC was 81%. Of the 104 patients, 5 (4.8%) and 9 (8.6%) died of unrelated causes and renal cancer, respectively. CONCLUSIONS: Our series confirmed a favorable outcome for the CRCC subtype with little local aggressiveness and a low propensity for progression and death from cancer. PMID- 15833509 TI - Outcomes of hand-assisted laparoscopic nephroureterectomy for managing upper urinary tract transitional cell carcinoma--China Medical University Hospital experience. AB - OBJECTIVES: To report our experience with hand-assisted laparoscopic nephroureterectomy (HALNU) to treat upper urinary tract transitional cell carcinoma (TCC). In addition, we report the treatment of 4 patients with upper urinary tract TCC and synchronous superficial urinary bladder TCC by HALNU and simultaneous transurethral resection of bladder tumor (TURBT). METHODS: We retrospectively reviewed 33 patients who had undergone HALNU. Recovery was evaluated according to the Eastern Cooperative Oncology Group performance status. Four patients had concomitant superficial urinary bladder cancer and underwent simultaneous TURBT at the beginning of surgery. We compared our data with those of our open surgery group and previously published data. RESULTS: Partial recovery had occurred by 1 week and complete recovery by 4 weeks postoperatively. Conversion to open surgery was required in 2 (6%) of the 33 patients. The complication rate was 24% (8 of 33) without any mortality. The recurrence rate of urinary bladder TCC was 6% (2 of 33). Both patients with recurrent tumor were treated successfully by TURBT and bacille Calmette-Guerin instillation. All 4 patients with synchronous superficial urinary bladder TCC had undergone simultaneous transurethral resection of bladder tumor, and all 4 were disease free at the last follow-up visit. CONCLUSIONS: Hand-assisted procedures are appropriate for surgeons with limited experience with laparoscopic surgery. In our study, the convalescence of patients was excellent, and the postoperative results were compatible with those of previous studies. To treat upper urinary tract TCC with synchronous urinary bladder TCC, HALNU and TURBT together seems to be a safe and efficient technique. PMID- 15833510 TI - Prognostic value of lymphovascular invasion in transitional cell carcinoma of upper urinary tract. AB - OBJECTIVES: To elucidate the prognostic significance of lymphovascular invasion (LVI) in patients with upper tract transitional cell carcinoma. METHODS: Of 86 patients with upper tract transitional cell carcinoma who underwent nephroureterectomy with bladder cuff (95%) or parenchymal-sparing (5%) surgery from 1991 to 2002, and who met our inclusion criteria, the data of 73 were available for pathologic review of LVI. The mean patient age was 59.1 years, and the median follow-up was 42.3 months. Using univariate and multivariate analyses, we determined the influence of multiple prognostic factors, including age, sex, tumor stage (T or N), tumor grade, and LVI, on the 5-year disease-specific and recurrence (local recurrence or distant metastasis)-free survival rates. We generated 5-year disease-specific and recurrence-free survival curves in terms of LVI in patients without lymph node involvement. RESULTS: The overall 5-year disease-specific and recurrence-free survival rate was 88% and 75%, respectively (n = 73). In univariate analysis, T stage, grade, and LVI significantly affected both survival rates. N stage was significant for 5-year recurrence-free survival. In multivariate analysis, LVI was the only significant predictor of recurrence free survival, and no factor was significant for disease-specific survival. Of 10 patients with positive lymph nodes, 7 had LVI. In patients without lymph node involvement or Stage T4 disease (Ta-T3N0M0, n = 62), the 5-year disease-specific and recurrence-free survival rate was 98% and 94%, respectively, in the absence of LVI and 70% and 60%, respectively, in the presence of LVI (P = 0.0005 and P = 0.0007, respectively). CONCLUSIONS: LVI is an independent prognostic factor for recurrence-free survival in transitional cell carcinoma of the upper urinary tract. Because LVI is strongly associated with a poorer prognosis, systemic adjuvant therapy should be considered in the presence of LVI, even if the lymph nodes are not involved. PMID- 15833511 TI - Do vascular, lymphatic, and perineural invasion have prognostic implications for bladder cancer after radical cystectomy? AB - OBJECTIVES: To elucidate the respective prognostic implications of vascular, lymphatic, and perineural invasion noted on the pathologic analyses of radical cystectomy specimens. Controversy still exists on whether the pathologic features of vascular, lymphatic, and perineural invasion have any role as prognostic indicators for bladder cancer. METHODS: A retrospective review of 125 patients with bladder cancer treated with radical cystectomy was conducted. Patients who received either preoperative or postoperative chemotherapy, those with lymph node metastasis confirmed on postoperative pathologic analysis, those who did not undergo standard bilateral pelvic lymph node dissection, and those receiving palliative treatment were excluded from the study. RESULTS: The mean patient age was 62.5 years (range 39 to 84), and the median follow-up duration was 41.0 months (range 1 to 146). Vascular, lymphatic, and perineural invasion was present in 8.8%, 20.8%, and 8.8% of specimens, respectively, and 28% of patients had at least one of the three pathologic features in the specimen. Univariate analysis revealed that vascular invasion, lymphatic invasion, and perineural invasion were significant prognostic predictors of overall survival. However, only the tumor stage and vascular invasion proved to be independent prognostic predictors of disease-specific survival on multivariate analysis. CONCLUSIONS: The results of the present study have shown that the pathologic tumor stage and the pathologic presence of vascular invasion are independent prognostic predictors for disease specific survival in patients who have undergone radical cystectomy for bladder cancer. Additional study should be performed on the prognostic implications of lymphatic and perineural invasion. PMID- 15833512 TI - Preservation of prostate during radical cystectomy: evaluation of prevalence of prostate cancer associated with bladder cancer. AB - OBJECTIVES: To estimate the frequency and characteristics of prostatic lesions discovered incidentally in radical cystoprostatectomy specimens and to determine whether any factors would allow for the detection of prostate cancer preoperatively. METHODS: A total of 100 radical cystoprostatectomy specimens with orthotopic bladder reconstruction were performed for malignant bladder disease between 1990 and 2000. The mean patient age at surgery was 62 +/- 8 years (range 32 to 75). Digital rectal examination and prostate-specific antigen (PSA) assay were done routinely before surgery. During the 10-year study period, the same pathologist examined the prostatic tissues from radical cystoprostatectomy specimens using McNeal's technique on fine slices every 2.5 mm. RESULTS: The overall incidence of prostate cancer discovered incidentally in radical cystoprostatectomy specimens was 51%, of which 29% were microcancers (volume less than 0.5 cm3) and 22% were significantly larger (volume 0.5 cm3 or more). The mean Gleason score was 6. Of the tumors, 24% could be considered "clinically nonsignificant" (less than 0.5 cm3 and Gleason score less than 7). The mean preoperative PSA level was 4.13 +/- 1.36 ng/mL. Of 66 patients with a PSA level of less than 4 ng/mL (mean PSA 1.5 +/- 0.8) and a normal digital rectal examination before surgery, 50% had prostate cancer, of which 69% were microcancers. CONCLUSIONS: The prevalence of prostate cancer (51%) in our series is among the highest in published reports. Furthermore, our results stress that currently no factors are available to enable the detection of "clinically significant" prostate cancer preoperatively. PMID- 15833513 TI - Observed effect of age and body mass index on total and complexed PSA: analysis from a national screening program. AB - OBJECTIVES: To assess the effect of aging and body mass index (BMI) on both serum total prostate-specific antigen (tPSA) and complexed PSA (cPSA) from the results of the 2003 Prostate Cancer Awareness Week national screening database. METHODS: A total of 12,943 men were screened for prostate cancer. Of these, 4458 men had simultaneous measurements of tPSA and cPSA. Height and weight were used to calculate the BMI. The subjects were grouped by decade of age and categorized as either obese or nonobese with a cutoff BMI of 30 kg/m2. The rates of increase in mean tPSA and cPSA between consecutive decades were compared. The relationship among BMI, PSA, and cPSA was also analyzed using multivariate regression analysis. RESULTS: The median age of those screened was 60 years. The median PSA and cPSA was 1.10 and 0.69 ng/dL, respectively. Both serum tPSA and cPSA levels showed an exponential increase with advancing age. The ratio of cPSA/tPSA remained relatively constant at 0.6. Compared with the nonobese cohort, men with a BMI of 30 kg/m2 or more had significantly lower tPSA and cPSA levels across all age groups (P <0.01). The overall odds ratio for all age groups was 1.8 (95% confidence interval 1.2 to 2.3). CONCLUSIONS: Both tPSA and cPSA levels increased with age. Across age groups, men with a greater BMI had significantly lower cPSA and tPSA levels. The clinical significance of age-adjusted cutoff levels for PSA in obese men for the diagnosis of prostate cancer should be explored further in future prospective clinical trials. PMID- 15833514 TI - Circulating tumor cells predict survival in patients with metastatic prostate cancer. AB - OBJECTIVES: To determine whether circulating tumor cells (CTCs) predict for survival in patients with metastatic prostate cancer (PCa) and to compare its prognostic abilities with other clinical factors. METHODS: Blood samples from 37 patients with metastatic PCa were analyzed for CTCs. CTCs were enriched from 7.5 mL blood using magnetic nanoparticles targeting the epithelial cell adhesion molecule and then fluorescently labeled. The samples were analyzed by multiparameter flow cytometry, and events with appropriate light scatter properties that were nucleic acid dye positive, cytokeratin positive, and CD45 negative were defined as CTCs. RESULTS: The number of CTCs found ranged from 0 to 8586 per 7.5 mL (mean 530 +/- 1887, median 5). A threshold of 5 or more CTCs per 7.5 mL of blood was used to evaluate the ability of CTCs to predict for overall survival. Of the 37 patients, 23 (62%) had 5 or more CTCs, with a median overall survival of 0.70 year compared with more than 4 years for those patients with fewer than 5 CTCs (log-rank P = 0.002, Cox hazards ratio 7.4). In the subset of 26 patients with hormone-refractory PCa, the presence of CTCs was the most significant parameter predictive of survival in univariate and multivariate analyses. CONCLUSIONS: In this pilot study, the presence of 5 or more CTCs in 7.5 mL blood was associated with poor overall survival in patients with metastatic PCa. PMID- 15833515 TI - Increased intratumoral expression of prostate secretory protein of 94 amino acids predicts for worse disease recurrence and progression after radical prostatectomy in patients with prostate cancer. AB - OBJECTIVES: To examine the impact of prostate secretory protein of 94 amino acids (PSP94) expression within the primary tumor on disease recurrence and progression using radical prostatectomy specimens from patients with adenocarcinoma of the prostate. METHODS: PSP94 immunohistochemistry was performed on 59 radical prostatectomy specimens. The degree of PSP94 expression was reported as either present (group 1) or absent (group 2). Clinical data, including survival outcome measures, were correlated with PSP94 expression. RESULTS: The time to disease progression for group 1 was shorter compared with that for group 2 (P = 0.042). Disease-free survival was also less in group 1 than in group 2 (P = 0.033). Multivariate analyses demonstrated that PSP94 expression was an independent prognostic factor for a shorter interval to disease progression (P = 0.046) and disease-free survival (P = 0.049). CONCLUSIONS: PSP94 expression in radical prostatectomy tumor specimens appears to be associated with worsened survival outcomes and may provide additional prognostic information. PMID- 15833516 TI - Intermittent androgen deprivation for biologic recurrence after radical prostatectomy: long-term experience. AB - OBJECTIVES: To analyze the oncologic results of intermittent androgen deprivation (IAD) for biochemical recurrence after radical prostatectomy (RP). METHODS: A total of 57 patients with biochemical recurrence after RP have been treated with IAD at our institution. The 57 patients were divided into two groups: group 1 comprised 29 patients who received salvage radiotherapy after RP; group 2 comprised 28 patients who did not receive salvage radiotherapy. Hormonal therapy during the first treatment phase consisted of an antiandrogen alone. This treatment was maintained for 3 months after the prostate-specific antigen (PSA) level had become undetectable and was then discontinued. Hormonal therapy was resumed when the PSA level exceeded 4 ng/mL; treatment was discontinued when the PSA level dropped to less than 1 ng/mL. RESULTS: The patients in group 1 had less favorable characteristics than those in group 2 in terms of pathologic stage and Gleason score. Overall, the median follow-up after starting hormonal therapy was 92 months (range 36 to 176). The percentage of each cycle that was spent "off" treatment decreased from 60% to 50%. During follow-up, 38.6% of patients required a luteinizing hormone-releasing hormone analog for nonresponse to the antiandrogen alone, and 15.8% experienced metastatic progression. The cancer specific mortality rate was 12.3%; all patients who died of prostate cancer were from group 1. The median interval between initiation of hormonal therapy and cancer-related death was 86 months. CONCLUSIONS: In our experience, IAD for biochemical recurrence after RP provided satisfactory long-term oncologic results. Our data suggest that IAD can be initiated with an antiandrogen alone. PMID- 15833517 TI - Intraoperative cell salvage during radical prostatectomy is not associated with greater biochemical recurrence rate. AB - OBJECTIVES: To evaluate the risk of long-term biochemical recurrence for patients who receive cell-salvaged blood. Radical retropubic prostatectomy (RRP) is historically associated with the potential for significant blood loss. Different blood management strategies include blood donation, hemodilution, preoperative erythropoietin, and intraoperative cell salvage (IOCS). Oncologic surgeons have been reluctant to use IOCS because of the potential risk of tumor dissemination. METHODS: We retrospectively analyzed an RRP database and compared those who did and did not receive cell-salvaged blood by baseline parameters, pathologic outcomes, and biochemical recurrence. We also stratified our patients according to the risk of recurrence. RESULTS: A total of 1038 patients underwent RRP between 1992 and 2003. Of these, 265 (25.5%) received cell-salvaged blood and 773 (74.5%) did not. The two groups had similar baseline characteristics. No differences were found between the two groups when compared by risk of seminal vesicle invasion or positive surgical margins. Those who received cell-salvaged blood had a lower risk of extraprostatic extension. The median follow-up for all patients was 40.2 months. The overall risk of biochemical recurrence at 5 years for those who did and did not receive cell-salvaged blood was 15% and 18%, respectively (P = 0.76). No significant differences were found in the risk of biochemical recurrence when patients were stratified according to low, intermediate, and high risk. CONCLUSIONS: IOCS is a safe and effective blood management strategy for patients undergoing RRP. The risk of biochemical recurrence was not increased for those who received cell-salvaged blood. Concerns about spreading tumor cells by way of IOCS would seem unwarranted. PMID- 15833518 TI - Systematic transperineal ultrasound-guided template biopsy of the prostate: three year experience. AB - OBJECTIVES: To review the technique of transperineal saturation prostate biopsy and to update our results on patients at high risk of prostate cancer. METHODS: A total of 210 men who met the study inclusion criteria underwent systematic transperineal ultrasound-guided template biopsy of the prostate. All patients had previously undergone at least one set of transrectal prostate biopsies and 170 (81%) had undergone two or more. The mean number of prostate cores obtained before the template biopsy was 17.4. A mean of 21.2 cores (range 12 to 41) were obtained at the template biopsy, depending on prostate size. The study inclusion criteria included prostate-specific antigen level of 10 ng/mL or greater, prostate-specific antigen velocity of 0.75 ng/mL per year or greater, or the presence of prostatic intraepithelial neoplasia and/or atypical small cell acinar proliferation on the previous biopsy. RESULTS: Adenocarcinoma was detected in 78 men (37%). Cancer was detected in the transition zone in 60 (77%) of these 78 men, including 36 (46%) in whom cancer was identified uniquely in the transition zone. Gleason sum 6 (range 3 to 9) was the most common biopsy grade. Thirty patients underwent radical prostatectomy, and 27 (90%) had pathologic Stage pT2 disease as the final pathologic stage. Complications from biopsy were limited to postprocedural urinary retention in 11% of patients. CONCLUSIONS: A systematic transperineal template biopsy provides uniform sampling of the entire prostate. This technique appears to enhance the identification of transition zone cancers not detected by previous transrectal prostate biopsy in patients at high risk of prostate adenocarcinoma. PMID- 15833519 TI - Impact of obesity on clinical outcomes in robotic prostatectomy. AB - OBJECTIVES: To assess the preoperative parameters and clinical outcomes of patients undergoing robotic laparoscopic radical prostatectomy with specific attention to the body mass index (BMI). Little is known about the impact of obesity (BMI greater than 30) on the clinical outcomes of patients undergoing radical prostatectomy. METHODS: The data of 100 men undergoing robotic laparoscopic radical prostatectomy between June 2002 and October 2003 were prospectively entered into a database. The standard clinical characteristics (eg, prostate-specific antigen, Gleason score) and perioperative and postoperative parameters were evaluated. Additionally, all were assessed preoperatively and postoperatively for American Urological Association symptom and bother scores, uroflowmetry, postvoid residual urine volume, and sexual function. RESULTS: Nineteen men were obese (BMI greater than 30) and 81 were not (BMI less than 30). The two groups had a similar need for transfusion, length of stay, and pathologic outcome. However, the obese men had poorer baseline urinary function (peak flow rate 13.9 versus 18.3 mL/s; voided volume 306 versus 454 mL; P < or =0.05) and sexual function (Sexual Health Inventory of Men score 14.1 versus 18.2; P < or =0.05). Obese men had significantly more complications (26.3% versus 4.9%; P = 0.01) and required more time to return to baseline activities (7 versus 4.3 weeks; P = 0.09) and urinary function. Finally, at 6 months, only 47% of obese patients versus 91.4% of nonobese patients had achieved pad-free urinary continence (P < or =0.001). CONCLUSIONS: In this study, obese patients had significantly worse baseline urinary and sexual function, had complications, and did not recover urinary function as quickly or as well as nonobese patients. Obese patients also demonstrated a strong trend toward a delay in recovery time. PMID- 15833520 TI - Atrophy in prostate needle biopsy cores and its relationship to prostate cancer incidence in screened men. AB - OBJECTIVES: To evaluate whether the incidence of atrophy reported on sextant biopsies is associated with subsequent prostate cancer detection and to obtain a more thorough analysis of the different categories and extent of atrophy, we performed a review of benign biopsy cores. METHODS: In the Rotterdam section of the European Randomized Study of Screening for Prostate Cancer, 4117 and 1840 men underwent sextant biopsy in the first and second screening round (4-year interval), respectively. Sextant biopsy was prompted by elevated prostate specific antigen levels. For review, randomly taken benign sextant biopsies (n = 202) with a follow-up of at least 8 years were chosen. RESULTS: Before review, atrophy was reported in the biopsies of 11.4% and 8.7% of the first and second round, respectively. The prostate cancer incidence during 8 years of follow-up after an initial diagnosis of atrophy was 10.4%, which was not significantly different than the 12.3% of cancers detected after a benign diagnosis without reference to atrophy. After review, the incidence of simple atrophy, post atrophic hyperplasia, and sclerotic atrophy in sextant biopsies was 91%, 47%, and 9%, respectively. Extensive atrophy was observed in 5% of biopsies. Only 2 men (4.7%) in the reviewed group had a subsequent diagnosis of prostate cancer in the 8 years of follow-up. Additionally, prostatic intraepithelial neoplasia was diagnosed in 3 men (7.0%) in the second screening round. CONCLUSIONS: Atrophy, especially its simple form, is a very common lesion in prostate biopsy cores (94%). Atrophy in an asymptomatic population undergoing screening was not associated with a greater prostate cancer or prostatic intraepithelial neoplasia incidence during subsequent screening rounds. PMID- 15833521 TI - Acute urinary retention after magnetic resonance image-guided prostate brachytherapy with and without neoadjuvant external beam radiotherapy. AB - OBJECTIVES: To evaluate the risk of acute urinary retention (AUR) and sequelae after urethra-sparing magnetic resonance imaging (MRI)-guided prostate brachytherapy. METHODS: Between 1997 and 2003, MRI-guided prostate brachytherapy was performed after external beam radiotherapy (n = 60) or as monotherapy (n = 188) following a diagnosis of low-risk prostate adenocarcinoma. The median prostate gland volume was 40 cm3 (range 16 to 184). Of the 248 patients, 88 (35%) had prostate gland volumes greater than 45 cm3. The median follow-up was 40 months. RESULTS: Of the 248 patients, 18 (7%) developed AUR. Of the 248 patients, 2% (3 of 160), 6% (3 of 49), 28% (8 of 29), and 40% (4 of 10) of patients experienced AUR if the prostate gland volume was less than 45, 45 to 60, 60 to 90, or greater than 90 cm(3), respectively (P <0.0001). Retention requiring intermittent self-catheterization resolved in one half of the patients by 2 weeks and had resolved in all patients by 6 weeks. At 1, 3, and 6 months after implantation, 100%, 57%, and 29% of patients who experienced AUR, respectively, used Flomax (P <0.0001). The mean Flomax dose used decreased from 0.9 mg at 1 month to 0.53 mg at 1 year. CONCLUSIONS: AUR after urethral-sparing MRI-guided prostate brachytherapy is volume dependent and is self-limited despite very large prostate gland volumes. PMID- 15833522 TI - Co-administration of low doses of intranasal PT-141, a melanocortin receptor agonist, and sildenafil to men with erectile dysfunction results in an enhanced erectile response. AB - OBJECTIVES: To evaluate the safety and pharmacodynamic effect of co administration of subtherapeutic doses of PT-141, a cyclic heptapeptide melanocortin analogue, and sildenafil to patients with erectile dysfunction. METHODS: Nineteen patients with erectile dysfunction who were responders to either Viagra or Levitra by self-report were given 25 mg sildenafil and 7.5 mg intranasal PT-141, 25 mg sildenafil and an intranasal placebo spray, and a placebo tablet and an intranasal placebo spray in a randomized cross-over design. Erectile activity in response to two 30-minute episodes of visual sexual stimulation was assessed by RigiScan during a 6-hour postdose period. RESULTS: The erectile response induced by co-administration of PT-141 and sildenafil was significantly greater than the response elicited by administration of sildenafil alone. Co-administration of PT-141 and sildenafil was safe and well-tolerated and did not result in new adverse events or adverse events that were increased in frequency or severity compared with monotherapy. CONCLUSIONS: Co-administration of intranasal PT-141 and a phosphodiesterase type 5 inhibitor may constitute a treatment alternative for patients in whom higher doses of a single therapy are not effective or well tolerated. PMID- 15833523 TI - Peyronie's disease with erectile dysfunction: penile modeling over inflatable penile prostheses. AB - OBJECTIVES: To evaluate, retrospectively, the impact of penile correction by modeling of the penis over an inflatable penile prosthesis and the subsequent improvement in erectile function. Advanced Peyronie's disease with severe penile curvature and poor quality erections presents a challenge to the urologist. METHODS: In our series, 46 patients with advanced Peyronie's disease and associated erectile dysfunction underwent insertion of an inflatable penile prosthesis between 1998 and 2003. Of the 46 patients, 28 (61%) underwent a standard modeling procedure; the other 18 patients (39%) did not need additional modeling, because their curvature was corrected by inflation of the prosthesis alone. Patients were evaluated postoperatively in the clinic, as well as by a postal questionnaire. RESULTS: Of the 46 patients, 44 were satisfied with the penile correction and 2 (4.4%) underwent removal of their prosthesis because of infection. These 2 patients underwent revision surgery; subsequently both prostheses had to be removed, one for severe pain and the other for urethral erosion. None of the patients underwent reoperation for additional straightening. Of the 44 patients with intact prostheses, erectile function significantly improved in 41 (93%). CONCLUSIONS: The results of our study have indicated that patients with severe Peyronie's disease and erectile dysfunction should be offered the choice of penile modeling over an inflatable penile implant to correct the curvature, as well as improve erectile function. PMID- 15833524 TI - Effects of intravesical chemotherapy and immunotherapy on semen analysis. AB - OBJECTIVES: To study the effect of intravesical chemo-immunotherapy on the sperm parameters of young patients. METHODS: Twelve young male patients with superficial transitional cell carcinoma, all younger than 40 years old at surgery, were included in this prospective study. The mean patient age was 34.8 years (range 22 to 40). Of the 12 patients, 8 had superficial transitional cell carcinoma, grade 2-3, and 4 had proven invasion to the lamina propria; 1 patient had accompanying carcinoma in situ. Accordingly, adjuvant intravesical treatment with either bacille Calmette-Guerin (BCG; 6 patients) or mitomycin C (6 patients) was indicated on the basis of the initial stage and grade. Sperm analysis was performed before bladder irrigation and subsequently 3 months after completion of intravesical therapy. RESULTS: All 12 patients had normal follicle-stimulating hormone and luteinizing hormone levels after surgery. All 12 patients had normal volume ejaculate, except for 1 who had undergone multiple prior transurethral tumor resections. Of the 6 patients who were treated with mitomycin C, only a few minor insignificant changes in the sperm quality were noted, and 2 of them later fathered healthy children. However, in 3 of the BCG-treated patients, remarkable changes in all sperm quality parameters were evident, with a statistically significant decrease in the sperm count (P = 0.0021). CONCLUSIONS: We suggest that potential adverse effects on spermatogenesis can be induced by intravesical therapy with BCG and that, consequently, routine pretreatment semen preservation should be considered as a precaution before instillation of intravesical BCG to prevent subsequent fertility difficulties in young men. PMID- 15833525 TI - Effect of age and grade on surgery for patients with varicocele. AB - OBJECTIVES: To assess the seminal characteristics, hormonal levels, and pathologic findings in patients of various age groups with varicocele before and after ligation to establish the presence of an aging effect in patients with varicocele on those characteristics. METHODS: The records were retrospectively evaluated for 70 infertile patients with azoospermia or oligospermia who underwent microsurgical left inguinal varicocele ligation. The mean +/- SD patient age was 33.9 +/- 5.5 years. Left varicocele was detected in 70 patients. We divided our patients into three groups according to age: group 1, 20 to 29 years old (n = 17); group 2, 30 to 39 years old (n = 41); and group 3, older than 40 years (n = 12). RESULTS: Before ligation, no significant differences were seen among the three groups in luteinizing hormonal level, prolactin level, estradiol level, right testicular volume, semen volume, and sperm concentration and sperm motility. Significant differences were seen in the follicle-stimulating hormonal level, testosterone level, and left testicular volume. After ligation, even in group 3, the sperm concentration and motility increased from 6.38 +/- 5.47 to 13.07 +/- 9.05 million/mL (P = 0.042) and from 28.42% +/- 23.22% to 39.92% +/- 22.06%, respectively. No significantly greater degree of improvement in semen characteristics was observed between grade II and grade III after varicocele ligation. CONCLUSIONS: The results of our study have shown that age is not a significant predictive factor of improvement in semen characteristics before ligation. It is reasonable to perform ligation to improve the semen characteristics in patients older than 40 years old. PMID- 15833526 TI - Effect of neonatal circumcision on penile neurologic sensation. AB - OBJECTIVES: To evaluate penile sensory thresholds in neonatally circumcised and uncircumcised men. METHODS: We evaluated 125 patients, 62 uncircumcised men and 63 neonatally circumcised men. All patients completed the Erectile Function domain of the International Index of Erectile Function questionnaire. Of the 125 patients, 29 (International Index of Erectile Function score of between 25 and 30) were placed in the functional group, and 96 in the dysfunctional group. The patients were tested on the dorsal midline glans of the penis (foreskin retracted). Quantitative somatosensory testing was performed and included vibration, pressure, spatial perception, and warm and cold thermal thresholds. RESULTS: In the functional group, t-test analysis demonstrated a significant (P <0.001) difference, with worse vibration and better pressure sensation for uncircumcised men. When controlling for age, hypertension, and diabetes, all t test significance was lost. In the dysfunctional group, circumcised men (49 +/- 16 years) were significantly younger (P <0.01) than uncircumcised men (56 +/- 13 years). For the dysfunctional group, t-test analysis also demonstrated worse vibration sensation for uncircumcised men (P <0.01). Again, when controlling for age, hypertension, and diabetes, all t-test significance was lost. CONCLUSIONS: We present a comparative analysis of uncircumcised and circumcised men using a battery of quantitative somatosensory tests that evaluate the spectrum of small to large axon nerve fiber function. Our study controlled for factors, including age, erectile function status, diabetes, and hypertension, that have been shown to alter neurologic testing. In our study of neonatally circumcised men, we demonstrated that circumcision status does not significantly alter the quantitative somatosensory testing results at the glans penis. PMID- 15833527 TI - Use of mobile extracorporeal shock wave lithotripter: experience in a pediatric institution. AB - OBJECTIVES: To present our results of the past 3 years using a mobile lithotripter in a pediatric institution. The low incidence of pediatric urinary calculi, along with the high cost of lithotripsy units, has limited the use of extracorporeal shock wave lithotripsy in pediatric institutions. METHODS: A retrospective cohort study was performed of all children who had undergone extracorporeal shock wave lithotripsy at two institutions from 1999 to 2003. All patients were treated with a Dornier Compact Delta mobile lithotriptor with a universal urologic table. RESULTS: A total of 88 patients (38 males and 50 females) underwent a total of 119 outpatient treatments. The mean patient age was 12 years, and the mean follow-up was 20 months. The mean stone size was 6 mm. An average of 1600 shocks per procedure was performed. Overall, 68% were stone free after one treatment, and 74% were stone free after one or two treatments. Of the patients with a solitary stone, 72% were stone free after one treatment and 78% were after one or two treatments. Fifteen patients had multiple stones treated simultaneously on the same side. Of these, 47% were stone free after one treatment and 53% after one or two treatments. Two patients subsequently required ureteroscopy and laser lithotripsy for obstructing ureteral calculi. CONCLUSIONS: The results of our study have shown that the new-generation mobile lithotripter is safe and effective in treating pediatric nephrolithiasis. The success rate was greater for solitary stones than for multiple stones treated at the same setting. Additional endoscopic procedures can be performed simultaneously on the same table. PMID- 15833528 TI - Management of complications after hypospadias repair. AB - OBJECTIVES: To review our experience with hypospadias complications (seen after 10%-15% of repairs) and to identify factors influencing outcome. METHODS: We reviewed the available medical records of 113 patients who underwent repeat operation for hypospadias complications. Of the 113 patients, 40% had undergone the initial repair at our institution (internal referral); 60% had undergone the initial repair elsewhere before referral (external referral). The variables potentially affecting outcome were reviewed, including the severity of the defect, concomitant disease, age at the initial operation and revisions, type of complications and treatment, and the number of revisions. Outcomes were compared on the basis of specialty and experience. RESULTS: Isolated hypospadias was present in 81% and other genitourinary abnormalities in 10%. External referral patients were older at the first revision (7.3 versus 4.2 years, P = 0.027). Complications included fistula (73%), stricture (12%), breakdown of repair (10%), and diverticulum formation (11%). Successful revision was independent of the initial defect. The first, second, and third revision was successful in 77%, 64%, and 67% of patients, respectively. The cumulative success rate was 77%, 92%, and 97% after each respective repair attempt. The success of the repair was independent of the patient's age at the initial operation/revision and of the interval from the initial repair to reoperation. Internal referral and external referral patients had similar results. Specific experience with the repair of hypospadias complications correlated with a successful outcome (P <0.001). CONCLUSIONS: Complications after hypospadias repairs are common, with fistula accounting for approximately 75%. The outcome in our series was independent of hypospadias severity, patient age at repair, number of revisions, stent use, and referral status. Repairs performed by an experienced pediatric urologist were associated with improved outcomes (P <0.001). PMID- 15833529 TI - Modified laparoscopic radical nephroureterectomy with transurethralexcision of intramural ureter. AB - We report a modified technique for laparoscopic nephroureterectomy that results in complete removal of the specimen, including a cuff of bladder mucosa, while avoiding tumor cell spillage. A modified lithotomy position allows simultaneous laparoscopic and transurethral access without repositioning. Contraindications are juxtavesical or intramural ureteral tumor. PMID- 15833530 TI - Correcting congenital concealed penis: new pediatric surgical technique. AB - INTRODUCTION: Correction of the congenital concealed penis is a difficult surgical challenge. Many techniques have been described for its treatment that differ according to the incision lines and covering techniques, but the results have generally been disappointing. We report a novel surgical technique for unfurling the inner prepuce and outer penile skin and thus obtaining sufficient penile skin coverage. TECHNICAL CONSIDERATIONS: Surgical techniques for correcting the concealed penis are aimed at freeing the penis from any tethering or webbing, providing adequate penile skin coverage and fixing the proximal penile skin to the underlying fascia to prevent the body of the penis from retracting and detaching from its overlying skin. The unfurling method we describe here consists of dividing the outer penile skin to the penile base bilaterally, with ventral and dorsal incisions of the inner prepuce in the midline, perpendicular to the incision of the outer skin. CONCLUSIONS: This method is simple and provides a good blood supply to the flap. It appears to show good subjective and objective results, and no serious complications have developed. PMID- 15833531 TI - Magnetic resonance cystography: novel imaging technique for evaluation of vesicoureteral reflux. PMID- 15833532 TI - Successful embolization of symptomatic renal artery aneurysm in solitary kidney. PMID- 15833533 TI - Intraperitoneal drop metastasis after radiofrequency ablation of pararenal tumor recurrences. AB - We report the case of a 66-year-old woman who developed intraperitoneal drop metastasis after radiofrequency ablation of recurrent renal cell carcinoma. This patient had undergone multiple nephron-sparing procedures, including radiofrequency ablation to treat local recurrences. At routine follow-up, she was noted to have two intraperitoneal drop metastases, one in the right paracolic gutter and the other in the cul-de-sac of Douglas. The metastases were treated with complete surgical resection. To our knowledge, this is the first reported case of drop metastasis after radiofrequency ablation, drawing attention to the potential complications of this new treatment modality. PMID- 15833534 TI - Delivery of dead fetus from inside urinary bladder with uterine perforation: case report and review of literature. AB - The incidence of cesarean sections has increased radically worldwide in the second half of the 20th century, especially in developing countries. Generally, a trial of labor after a cesarean section is safely accepted. However, a trial of labor may lead to serious complications, including unpredicted ones, such as rupture of the uterus during labor with concomitant injury to the bladder. We report the delivery of a dead fetus through the urinary bladder during labor. PMID- 15833535 TI - Isolated vasculitis of the bladder. AB - We describe a case of isolated vasculitis involving the bladder that presented with severe irritative voiding and hematuria. This case presented a diagnostic problem, because malignancy was suspected, but not confirmed by biopsies. Once the vasculitis was diagnosed, secondary systemic causes were excluded, and successful treatment was subsequently undertaken using systemic steroids. It is important to differentiate this rare pathologic entity from the much more frequent tumors of the bladder because the treatment is medical rather than surgical. PMID- 15833536 TI - Urothelial carcinoma featuring lipid cell and plasmacytoid morphology with poor prognostic outcome. AB - An aggressive urothelial carcinoma classified as lipid cell variant with plasmacytoid features was diagnosed in a 67-year-old man. His overall survival was 18 months after limited surgery followed by chemotherapy. The histopathologic features were the same in the recurrences. PMID- 15833537 TI - Multiple retained needles: an unusual complication of intracavernous self injection. AB - Intracavernous self-injection of pharmacologic agents is an effective method of treatment of erectile dysfunction. Intracorporeal needle breakage is a rare complication of intracavernosal self-injection. We report an unusual case of multiple retained needles in a 44-year-old man who presented with penile edema after intracavernous self-injection complicated by needle breakage. The patient was treated with nonoperative therapy and observed on a course of antibiotics to which he responded favorably. He continued to have good erectile function. We also present a review of the reported data and recommend a treatment strategy for the management of retained intracorporeal needles. PMID- 15833538 TI - Renal epithelioid angiomyolipoma associated with hyperprolactinemia. AB - We report a unique case of a 42-year-old woman with a renal epithelioid angiomyolipoma associated with elevated serum prolactin levels and presenting with galactorrhea. Magnetic resonance imaging of the brain showed no pituitary abnormality, and abdominal imaging demonstrated a large right renal mass. Pathologic analysis of the radical nephrectomy specimen revealed an epithelioid angiomyolipoma. She had an uneventful postoperative recovery with complete normalization of her serum prolactin level and resolution of galactorrhea. To our knowledge, the association between epithelioid angiomyolipoma and hyperprolactinemia has never before been reported. PMID- 15833539 TI - Suprapubic electroresection of prostate in three patients with benign prostatic hyperplasia and previous surgery for long urethral stricture. AB - The treatment of patients presenting with severe symptoms of obstruction due to benign prostatic hyperplasia and a history of previous surgery for long urethral stricture is still a matter of discussion. We report on 3 patients in whom resection of the prostate was performed using a dilated cystostomy approach under spinal anesthesia. All men had undergone dorsal onlay urethroplasty for long urethral stricture before prostate resection. The good outcome demonstrated the feasibility and safety of prostate resection using a suprapubic approach, which may become a treatment option in this group of patients. PMID- 15833540 TI - Compression of kidneys in Erdheim-Chester disease of retroperitoneum: Open surgical approach. AB - Erdheim-Chester disease is a rare systemic non-Langerhans cell histiocytosis. We report the first case of surgical treatment of severe compression of renal parenchyma by retroperitoneal masses in a 61-year-old male patient with progressing renal failure. After 3 years of follow-up, we have concluded that the open surgical approach is an option in the management of renal complications in Erdheim-Chester disease. PMID- 15833541 TI - Synchronous inverted papilloma of bladder and renal pelvis. AB - Inverted papilloma of the urinary tract is a rare tumor that consists of uniform urothelial cells with an inverted configuration and a normal urothelial lining. Despite several published reports, the prognostic significance and the potential biologic behavior remain uncertain. We detail a case of synchronous inverted papilloma of the urinary bladder and the renal pelvis and review the published data. PMID- 15833542 TI - Epididymal tuberculosis: case report and review of the literature. AB - The incidence of tuberculosis (TB) is increasing worldwide, with more than 20% of cases exhibiting extrapulmonary manifestations. The genitourinary tract is the most common site of extrapulmonary TB. Epididymal TB, although rare, may be the initial location of infection and may cause infertility. The diagnosis depends on culture of the organism but has been facilitated by the development of urine polymerase chain reaction analysis. Treatment for TB remains the combination of three or four anti-TB drugs for 6 to 9 months. Genitourinary TB remains relatively rare in the United States and requires a high index of suspicion to make the diagnosis. PMID- 15833543 TI - Rare cause of ureteral stenosis more than 10 years after kidney transplantation. AB - Late renal graft failure is in most cases due to a chronic allograft nephropathy. In this report, we present a case in which a surgical complication led to ureteral stenosis more than 10 years after transplantation. The patient developed slowly deteriorating renal function and ultimately progressive hydronephrosis. At surgical exploration, the ureter was found to perforate the wall of the small bowel before entering the bladder. We successfully performed ureter reimplantation to restore the outflow of the kidney. PMID- 15833544 TI - Urethral strictures incident to bicycle motocross racing. AB - A dramatic shift from traditional team to alternative or "extreme" sports has given rise to a new generation of nontraditional athletes and sports-related injuries in the pediatric population. We present a case of 2 brothers who developed urethral strictures believed incident to BMX racing. We address current demographics and the general presentation and course of treatment to aid both the pediatric urologist and the general practitioner in prompt and proper diagnosis. PMID- 15833545 TI - Wegener's granulomatosis presenting as renal mass: a case for nephron-sparing surgery. AB - Wegener's granulomatosis (WG) is a systemic disease well recognized by physicians. The upper and lower respiratory symptoms associated with the disease are easily recognized; however, its presentation as a solid renal mass is underappreciated. We present a case of a 61-year-old man who presented with a 5.2 cm renal mass diagnosed as WG after nephron-sparing surgery. The patient presented with mild symptoms suggestive of WG, but the antineutrophil cytoplasmic antibody test was negative. The renal mass and concerns about possible malignancy obscured the diagnosis of WG. This case illustrates that WG should be considered in the differential diagnosis of solid renal masses. PMID- 15833546 TI - Percutaneous injection of fibrin glue for persistent nephrocutaneous fistula after partial nephrectomy. AB - We report a case of persistent urinary leak of nearly 4 months' duration after open surgical partial nephrectomy. The urinary leak was refractory to ureteral stenting, urethral catheter placement, and ureteroscopic fulguration. Fibrin glue was injected percutaneously under fluoroscopic guidance into the nephrocutaneous fistula tract, which resulted in its prompt and complete resolution. PMID- 15833547 TI - Pitfall in renal cyst surgery: serous cystadenoma of pancreas mimicking renal cyst. AB - Pancreatic lesions, particularly cysts, can simulate various diseases. We report a case of a 43-year-old woman with a large, symptomatic, retroperitoneal cyst misdiagnosed as a "renal cyst." During the retroperitoneoscopic marsupialization, the correct diagnosis of a pancreatic cyst was made, leading to an open pancreas tail resection. Histologic evaluation revealed serous cystadenoma. Especially in large retroperitoneal cysts on the left side, the correct diagnosis of a pancreatic cyst can be difficult. PMID- 15833548 TI - Prostate-specific antigen decline after gonadotropin-releasing hormone antagonist withdrawal in androgen-independent prostate cancer. AB - A 66-year-old man with androgen-independent prostate cancer was treated with abarelix, a gonadotropin-releasing hormone antagonist, for 20 weeks in an experimental protocol. He did not respond to therapy, but his serum prostate specific antigen level dropped from 15.8 ng/mL to a confirmed 0.8 ng/mL after abarelix was stopped. His prostate-specific antigen level did not return to greater than 15.8 ng/mL for 14 months. This is the first report of a withdrawal response after therapy with a gonadotropin-releasing hormone antagonist, a new class of agents for prostate cancer. Additional observations are needed to determine whether this is an isolated case or a harbinger of a more common phenomenon. PMID- 15833549 TI - Medial thigh pain: neurology or urology? AB - Flank pain, irritative urinary symptoms, and hematuria are traditional symptoms of urolithiasis. We report a case of urolithiasis in a neobladder in a patient with a chief complaint of medial thigh pain. PMID- 15833550 TI - Influence of estradiol pretreatment on antimuscarinic action of oxybutynin in rat detrusor muscle. AB - OBJECTIVES: To investigate the antimuscarinic effect of oxybutynin in the rat detrusor muscle after estrogen pretreatment because, to our knowledge, no study has been done on the interaction of estrogen with antimuscarinic drugs. Estrogen has been shown to affect muscarinic receptors in the detrusor muscle of animals. In addition, oxybutynin has been shown to block muscarinic receptors in the bladder. METHODS: Estradiol benzoate (150 microg/kg) or saline was given subcutaneously to virgin female Wistar albino rats (n = 6, each group) for 10 consecutive days. On the 11th day, isolated detrusor muscle strips were taken, and acetylcholine (ACh)-induced contractions were evaluated in the absence or presence of oxybutynin (10 and 100 nM). RESULTS: ACh induced concentration dependent contractions in the detrusor muscle. In the estradiol-pretreated group, the maximum of the ACh-induced contractions was diminished compared with that in the control group (P < 0.05). Oxybutynin (10 and 100 nM) inhibited ACh-induced contractions competitively (pK(B) 8.85). In the estradiol-pretreated group, the concentration-response curve to ACh was shifted further to the right in the presence of oxybutynin (100 nM). CONCLUSIONS: We have demonstrated for the first time that oxybutynin further inhibits ACh-induced and muscarinic receptor mediated contractions in rat detrusor muscle after pretreatment with estrogen. PMID- 15833551 TI - Gene gun particle encoding preproenkephalin cDNA produces analgesia against capsaicin-induced bladder pain in rats. AB - OBJECTIVES: To evaluate the efficacy of gene therapy using a gene gun or direct injection for the transfer of human preproenkephalin (PPE) plasmid cDNA using a capsaicin-induced bladder pain model in rats. Opioid peptides play an essential role in the modulation of micturition reflex and control of inflammatory pain. PPE is one such precursor molecule. METHODS: Human PPE cDNA was cloned into a modified pCMV plasmid and delivered into the bladder wall of adult female rats by direct injection or gene gun. At 4 and 7 days after gene therapy, continuous cystometrograms were performed under urethane anesthesia by filling the bladder (0.08 mL/min) with saline, followed by 15 muM capsaicin. Immunohistochemical staining was used to detect enkephalins in the bladder after PPE cDNA transfer. RESULTS: The intercontraction interval was decreased after intravesical instillation of capsaicin (65.0% and 63.1% decrease) in the control group or direct PPE gene injection group, respectively. However, the gene gun-treated group showed a significantly reduced response to capsaicin instillation at day 4 and day 7 (intercontraction interval 16.2% and 42.8% decrease, respectively). This analgesic effect was reversed by intravenous naloxone, an opioid antagonist (5 mg/kg). Increased enkephalin immunoreactivity in the bladder was observed in the gene gun-treated group at day 4, which was reduced at day 7. CONCLUSIONS: The PPE gene can be effectively transferred and suppress the nociceptive response in the bladder using the gene gun method. These results support potential clinical application of PPE gene gun delivery system for the treatment of bladder pain and other types of visceral pain. PMID- 15833552 TI - Robotic-assisted vasovasostomy: a two-layer technique in an animal model. AB - OBJECTIVES: To assess the feasibility of a multilayered robotic-assisted vasovasostomy (RAVV) in a rabbit model. Microscope-assisted vasovasostomy (MAVV) is a technically challenging procedure. Robotics may be a surgical adjunct that helps overcome the microsurgical challenges, which include fine suture, delicate instruments, and tremor. A recent survey revealed that most urologists use a multilayered technique for vasovasostomies. METHODS: A surgeon performed eight vasovasostomies with 10-0 suture and a two-layer technique using an in vivo rabbit model-four were MAVV using conventional microsurgical instrumentation and four were RAVV using the da Vinci robot. Performance measures and adverse haptic events were recorded. Patency was evaluated by passing a 2-0 Prolene suture through the anastomoses. RESULTS: The mean operating time for the total procedure and for the mucosal layer only was longer for RAVV than for MAVV (75 versus 42 minutes, P = 0.03 and 38 versus 23 minutes, P = 0.03, respectively). The needle passes required for the mucosal layer and the number of mucosal and muscularis sutures were similar in both groups (9.5 versus 8.8 passes, P = 0.34; 4 versus 4, P >0.99; and 7 versus 6.3, P = 0.2, respectively). Unlike MAVV, no tremor was appreciated during RAVV. No adverse haptic events were observed in either group. All anastomoses were patent, and all rabbits were free of any crush injury. CONCLUSIONS: A multilayered RAVV can be performed in an in vivo rabbit model. Although it was associated with increased operative times, the absence of adverse haptic events and comparable patency rates continue to suggest a role for robotics in microsurgery. PMID- 15833553 TI - Expression of matrix metalloproteinase-10 in human bladder transitional cell carcinoma. AB - OBJECTIVES: To analyze matrix metalloproteinase-10 (MMP-10) expression in transitional cell carcinoma (TCC) of the bladder, evaluate the correlations between MMP-10 protein expression and clinicopathologic parameters, and address the viability of MMP-10 as a therapeutic target for TCC. MMP-mediated degradation of the extracellular matrix is an important factor in the pathogenesis of tumorigenesis and metastasis. METHODS: Using immunohistochemistry, the expression of MMP-10 was assessed using both tissue microarrays and whole sections of archival tissue specimens representative of all grades and stages of human bladder TCC (n = 60). MMP-10 expression was also assessed in histologically normal human bladder tissue (n = 10). The immunostaining results for MMP-10 expression were examined for correlations with tumor grade and stage. RESULTS: Unlike most MMPs, MMP-10 was localized primarily in the tumor mass as opposed to the tumor stroma and was detectable in all grades and stages of TCC. Significantly greater levels of MMP-10 protein were observed in superficial (pTa, pT1; n = 38) tumors than in normal bladder tissue (P = 0.01). In contrast to the proposed role of MMPs in tumor invasion, no significant difference was observed between muscle-invasive tumors (pT2 or worse; n = 22) and histologically normal bladder tissue (P = 0.50). MMP-10 expression showed no significant correlation with tumor grade. CONCLUSIONS: The data from our study showed that, unlike most MMPs, MMP-10 was not associated with tumor aggression or invasion. Our results suggest that MMP-10 protein levels are significantly greater in the earlier stages of TCC development. PMID- 15833554 TI - Human embryoid body-derived stem cells in co-culture with bladder smooth muscle and urothelium. AB - OBJECTIVES: Human pluripotent stem cells have excellent regenerative abilities, making them attractive in bladder regeneration. Porcine small intestinal submucosa (SIS) is an established matrix well-suited to the urinary tract. We evaluated the ability of human embryonic germ (hEG) cell-derived stem cells to form a composite graft on SIS, grown alone or in combination with human bladder smooth muscle cells (SMCs) or urothelial (URO) cells. METHODS: The hEG cell derived cell line SDEC had the best-fit profile for smooth muscle, as determined by its gene and protein expression. SDEC cells were seeded on SIS, either on its serosal or mucosal surface, and cultured for 7 or 14 days. Co-cultures of stem cells with URO cells or SMCs were also assessed under similar conditions. The grafts were analyzed by histologic examination for cell growth, morphology, and matrix penetration. RESULTS: SDEC cells grew in a monolayer on SIS, with a marked increase in three-dimensional growth when co-cultured with URO cells or SMCs. Penetration of the matrix was evident when seeded on the rough surface of the SIS, progressing with time. Stratification of the cell layers occurred on the smooth side of the SIS. CONCLUSIONS: This is the first description of hEG cell derived stem cells in co-culture with bladder cells. hEG cell-derived stem cells grow well on SIS even when seeded at low concentrations in the presence of bladder cells (URO cells or SMCs). These composite grafts will be ideal to evaluate the in vivo functional characteristics of stem cells using an animal model of bladder regeneration. PMID- 15833555 TI - Human embryoid body-derived stem cells in bladder regeneration using rodent model. AB - OBJECTIVES: To evaluate the capability of a human embryonic germ (hEG) cell derived cell line (SDEC), previously characterized in our laboratory, seeded on porcine small intestinal submucosa (SIS) to regenerate the injured rat bladder. METHODS: Fluorescent-labeled SDEC cells seeded on SIS for 8 days in vitro were used as bladder grafts in rats. A total of 30 congenitally athymic rats (six groups of 5 rats each), underwent partial cystectomy and replacement with plain SIS (groups 1 to 3) or cell-seeded SIS (groups 4 to 6). The rats were sacrificed after 7 (groups 1 and 4), 14 (groups 2 and 5), and 28 (groups 3 and 6) days. The bladders were analyzed by histopathologic examination and fluorescence microscopy. RESULTS: No graft rejection or diminution in bladder capacity occurred. Plain SIS implants had multiple calcareous deposits, not seen with the cell-seeded implants. Macroscopically, at 7 days, the grafts were healed with a cellular lining on the luminal aspect in groups 4 to 6. Microscopically, the rat bladder was completely regenerated 28 days after stem cell-seeded SIS implantation. Labeled stem cells were identified throughout the graft and contributed significantly to bladder regeneration. CONCLUSIONS: The results of this study have demonstrated the successful replacement of a bladder defect in a rat model using hEG cell-derived cells seeded on SIS grafts. Longer term analysis of these bladder grafts will allow evaluation of function, cell migration, and differentiation processes of human stem cells. PMID- 15833557 TI - Clinical pathology in veterinary geriatrics. AB - Alterations in an individual analyte rarely provide an indication of the initiating circumstances that caused the abnormality. It is obvious from the previous discussion that multiple organs or organ systems can cause abnormal results in the same analyte. This fact underscores the importance of evaluating a biochemical profile in an integrated fashion, relating abnormalities of a particular analyte with the rest of the profile as well as with the signalment, history, and physical findings in the patient. Furthermore, assessment of abnormalities should be approached with some degree of skepticism because they may not be indicative of an actual disease. PMID- 15833558 TI - Geriatric pharmacology. AB - When faced with the geriatric dog or cat, the practitioner should consider the following: 1. Avoid using any drugs at all unless there are definite therapeutic indications. If the patient has some degree of renal insufficiency, try to select drugs that are hepatically metabolized and excreted in bile rather than eliminated by the kidneys (eg, doxycycline, tolfenamic acid). If hepatic insufficiency is present, select drugs that do not undergo metabolism before renal excretion (eg, penicillins, cephalosporins). 2. If therapeutic drug monitoring is available, tailor the drug dosage regimen to that specific patient (eg, phenobarbital, digoxin, amino-glycosides). 3. If therapeutic drug monitoring is unavailable, determine if there are clinically proven adjusted dosage regimens for specific drugs. The package insert on human pharmaceutics often gives guidelines for adjusting dosages in geriatric patients. 4. If the drug has not been sufficiently studied to have dosage adjustment recommendations, determine if there is sufficient information about its kinetics to estimate the proper drug dose in a geriatric patient. Some general guidelines for commonly used drugs in geriatric veterinary patients are provided in Table 1. In general, if the Vd changes in your patient, change the dose. If the elimination half-life changes, change the dosing interval. 5. Carefully monitor treated patients for signs of efficacy and toxicity. PMID- 15833559 TI - Anesthesia for geriatric patients. AB - Choosing the best anesthetic agents for each geriatric animal does not in itself ensure a successful outcome. Aggressive, careful, vigilant monitoring during the anesthetic and recovery periods is required to detect and correct alterations in homeostasis that may develop during the perianesthetic period. With appropriate preoperative screening, informed choice and judicious dosing of anesthetics, and careful monitoring and supportive care, the risk of anesthesia in geriatric animals can be greatly reduced. PMID- 15833560 TI - Early detection of renal damage and disease in dogs and cats. AB - Renal damage and disease can be caused by acute or chronic insults to the kidney. Acute renal damage often results from ischemic or toxic insults and usually affects the tubular portion of the nephron. In contrast, chronic renal disease can be caused by diseases and/or disorders that affect any portion of the nephron, including its blood supply and supporting interstitium. Early detection of acute renal disease facilitates appropriate intervention that can arrest or at least attenuate tubular cell damage and the development of established acute renal failure. Similarly,early detection of chronic renal disease, before the onset of renal azotemia and chronic renal failure, should facilitate appropriate intervention that stabilizes renal function or at least slows its progressive decline. PMID- 15833561 TI - Geriatric heart diseases in dogs. AB - A discussion of the diagnosis and therapy of heart disease in an aged pet does not differ significantly from that in a pet of any age. Mitral regurgitation constitutes by far the most important geriatric heart disease, and the selection of drugs to treat heart disease of aging pets is based on identification of specific pathologic features (eg, atrial fibrillation, left atrial enlargement) for which each aspect of treatment (eg, diuretics, angiotensin-converting enzyme inhibitors, spironolactone) is specific. PMID- 15833562 TI - Liver disease in the geriatric patient. AB - Normal functioning of the liver does not seem to change significantly in dogs and cats as a result of age. Despite this, older dogs and cats are at greater risk for the development of liver disease. The diagnosis of liver disease is initiated by the veterinarian's suspicion that liver disease might be present, followed by the case history and a physical examination. The initial workup for the older dog or cat with suspected liver disease should begin with a CBC, serum chemistry profile, and urinalysis. This may be followed by a liver function test, radiographic or ultrasonographic imaging studies, hepatic fine-needle aspiration, and, ultimately, liver biopsy. PMID- 15833563 TI - Thyroid disorders in the geriatric patient. AB - Thyroid disorders are common in older pets. They often present a diagnostic challenge, and reaching a definitive diagnosis can be difficult or impossible in some cases. It is important for the veterinary practitioner to be familiar with the historical, physical examination, and clinicopathologic data findings in each of these diseases and to become comfortable with the treatment, monitoring, and prognosis associated with thyroid diseases in geriatric pets. PMID- 15833564 TI - Orthopedic problems in geriatric dogs and cats. AB - Senior dogs and cats with orthopedic injuries and diseases often require a treatment plan that differs from that of younger patients. Injured bone and soft tissues tend to heal more slowly in the geriatric patient. The older animal is likely to have a less competent immune system and may have compromised metabolic and endocrine function. Pre-existing musculoskeletal problems may make ambulation difficult for an animal convalescing from a new orthopedic problem. Special attention is often needed when treating these patients for fractures, joint instability, infection, and neoplasia. In general, issues that should be addressed in the geriatric patient include reducing intraoperative and anesthesia time, enhancing bone and soft tissue healing, return to early function, control of postoperative pain, physical therapy, and proper nutrition. PMID- 15833565 TI - Behavior problems in geriatric pets. AB - Aging pets often suffer a decline in cognitive function (eg, memory,learning, perception, awareness) likely associated with age-dependent brain alterations. Clinically, cognitive dysfunction may result in various behavioral signs, including disorientation; forgetting of previously learned behaviors, such as house training; alterations in the manner in which the pet interacts with people or other pets;onset of new fears and anxiety; decreased recognition of people, places, or pets; and other signs of deteriorating memory and learning ability. Many medical problems, including other forms of brain pathologic conditions, can contribute to these signs. The practitioner must first determine the cause of the behavioral signs and then determine an appropriate course of treatment, bearing in mind the constraints of the aging process. A diagnosis of cognitive dysfunction syndrome is made once other medical and behavioral causes are ruled out. PMID- 15833566 TI - Geriatric veterinary dentistry: medical and client relations and challenges. AB - Quality of life is an important issue for geriatric patients. Allowing periodontal disease, fractured teeth, and neoplasia to remain untreated decreases this quality of life. Age itself should be recognized; however, it should not be a deterrent to successful veterinary dental care. PMID- 15833567 TI - Nutrition for aging cats and dogs and the importance of body condition. AB - Before recommending a diet for a senior pet, a thorough nutritional evaluation should be completed. Although many middle-aged and older pets are overweight, a large percentage of geriatric cats and dogs have a low BCS. Approximately one third of cats older than 12 years of age may have a decreased ability to digest fat, whereas one in five may have a compromised ability to digest protein. Thus, appropriate diets for these two age groups may differ considerably. Mature (middle-aged) cats would likely benefit from a lower calorie food, whereas geriatric cats (>12 years of age) may need a highly digestible nutrient-dense diet. More than 40% of dogs between the ages of 5 and 10 years are overweight or obese. Such dogs may benefit from diets with lower fat and calories. Senior dogs also have an increased need for dietary protein, however. Therefore, healthy older dogs may benefit from diets with an increased protein-to-calorie ratio, providing a minimum of 25% of calories from protein. Common obesity-related conditions in dogs or cats include DM and OA. Diabetes differs between dogs and cats. Type I diabetes, common in dogs, seems to respond to fiber-enriched diets, whereas type II diabetes, common in cats, seems to benefit from high-protein and low-carbohydrate diets. OA, an inflammatory condition that occurs in approximately 20% of dogs, may benefit from weight management and nutrients that reduce the inflammatory responses, such as long-chain n-3 fatty acids. PMID- 15833568 TI - Senior and geriatric care programs for veterinarians. AB - Earlier detection allows earlier intervention, and thus improved treatment success. Senior profiling improves anesthetic safety by identifying hidden existing diseases and permitting the postponement of anesthesia or altering the anesthetic plan. Furthermore, pharmaceutic safety is increased through the detection of underlying diseases that may preclude the use of certain drugs or suggest new alternative treatments. Many dietary recommendations are based on disease diagnosis, making senior profiling an important dietary database. Finally, earlier disease management by means of improved anesthetic, pharmaceutic, and dietary recommendations offers our patients and clients the best medical management possible. PMID- 15833569 TI - Nicotine and other substance interaction expectancies questionnaire: relationship of expectancies to substance use. AB - Smoking and substance abuse co-occur at high rates and substance abusers are less likely to quit smoking than are smokers in general. Therefore, more information about the beliefs substance abusers have about the role of smoking in substance use and in recovery would be useful when designing interventions to impact smoking among substance abusing patients. The present study developed a Nicotine and Other Substance Interaction Expectancies Questionnaire (NOSIE) to investigate the expectancies held by substance abusers in treatment about the effects of smoking on substance use, the effects of substance use on smoking, smoking to cope with recovery, and receptivity to smoking cessation during substance abuse treatment. The 29 items were Likert-rated by 160 substance dependent patients in an inner-city residential substance abuse treatment program and participating in a larger study of smoking at this site. Four components were derived and reduced to a 20-item measure with good reliability. No differences by gender or age were found. On average, the patients reported that substance use almost always increases their smoking or urges to smoke but that smoking only increased substance use or urges about half of the time, that they use smoking to cope with urges to use substances about half of the time, and that they generally agreed that smoking cessation or treatment should be tried during substance abuse treatment and would not harm recovery efforts. Three of the scales correlated with smoking dependence while one scale correlated with drug use severity and heavy drinking days. The scale of receptivity to smoking cessation correlated significantly with measures of motivation and barriers and predicted 1-month smoking cessation outcomes. However, scale scores on smoking to cope with recovery did not significantly predict 3-month relapse to substance use. Implications for theory and clinical interventions with substance abusers who smoke were discussed. PMID- 15833570 TI - Women and cigarette smoking: does amount of weight gain following a failed quit attempt affect social disapproval? AB - This study examined the perception of social attractiveness and approval of smoking in a vignette of an American college female smoker after a failed quit attempt. It was hypothesized that she would be perceived more favorably when she was smoking to control her weight, compared to a non-weight related excuse during a failed quit attempt. The design was a 2 (gender: male vs. female)x4 (condition: control, 5, 10, 20 lb gain)x3 (participants' smoking status: never, former, or current smoker) factorial design with the Social Attraction Index and Perception of Smoking Index as dependent variables. Contrary to the hypothesis, perceptions of the female were not affected by her disclosed amount of weight gain. However, consistent with the literature on stigmatization of smokers, smoking participants perceived the female target as most socially attractive and nonsmokers perceived her to be the least attractive. PMID- 15833571 TI - Intentions to quit smoking change over short periods of time. AB - To assess the stability of intention to quit smoking, 115 US and Swedish smokers were randomized to complete Stage of Change (SOC) or ladder scales of intentions to quit at either 0, 7, 14 and 30 days or at 0 and 30 days in the absence of intervention. The four-assessment group had more progression in intention to quit than the two-assessment group. Depending on the measure, 12-17% of smokers changed their intention to quit over 7 days, 15-25% changed over 14 days and 17 34% changed over 30 days. Results were similar in Swedish and US participants and replicate the results of prior studies. We conclude intention to quit often spontaneously changes over short periods of time, especially with repeated testing. PMID- 15833572 TI - Mode effects for collecting alcohol and tobacco data among 3rd and 4th grade students: a randomized pilot study of Web-form versus paper-form surveys. AB - OBJECTIVES: The present study examined the feasibility and possible mode effects of using a Web-form versus a paper-form survey to collect alcohol and tobacco data from 3rd and 4th grade students. METHODS: A survey regarding alcohol and tobacco use and attitudes was administered to 280 3rd and 4th grade students attending a racially and economically diverse Midwestern urban school district in the spring of 2003. The sample was randomly assigned to complete a Web-form survey (n=137) or a hardcopy paper-form survey (n=143). RESULTS: The overall response rate was 87% and did not differ by survey mode. There were minimal differences between survey modes in data quality including substantive responses to alcohol and tobacco use. CONCLUSIONS: The findings of the present study offer preliminary evidence that Web-form surveys can be considered for collecting alcohol and tobacco use data among young children. Although there were few survey mode differences, our findings suggest more research is needed to examine possible measurement errors induced by computer-based approaches of data collection. PMID- 15833573 TI - Alcohol expectancies and drinking behaviors in Mexican American college students. AB - Research with non-Latinos has shown that alcohol expectancies ("the anticipated effects of alcohol use") are associated with drinking behaviors; however, prior studies have not fully examined such relations in Latino college students. This investigation examines positive alcohol expectancies and their association with contextual drinking and alcohol-related problem behaviors in a sample of 107 Mexican American college students (mean age=25; 54% female) who completed self report surveys. Results showed that students who endorsed expectancies of physical and social pleasure and increased social assertiveness were likely to engage in frequent heavy drinking and imbibe in less personal settings. Furthermore, gender (being male) and the expectancy of enhanced sexual performance and experience were associated with involvement in risk behaviors due to drinking. This study highlights the link between specific alcohol expectancies and alcohol-related problem behaviors in Mexican American college students. Intervention approaches aimed at modifying drinking expectancies in physical, social, and sexual domains could prove useful in this population. PMID- 15833574 TI - The role of curiosity in smoking initiation. AB - CONTEXT: Although advertising theories have long viewed curiosity as an intermediate goal to encouraging consumption of a product among previous nonusers, this variable is rarely discussed in psychological theories and its role in smoking uptake has not been addressed adequately. DESIGN AND SETTING: Using a longitudinal design, in 1999, we reinterviewed 12- to 15-year-old adolescent never smokers (N=2119; 970 committed never smokers, 1199 susceptible never smokers) 3 years after they responded to a population survey on tobacco use in California. RESULTS: Logistic regression showed curiosity and susceptibility to smoke were independently associated with increased future smoking in all never smokers. Committed never smokers reporting curiosity were more likely to become susceptible by follow-up than those not curious. A multivariate analysis of committed never smokers from the cross-sectional survey identified best friends who smoke and receptivity to tobacco advertising and promotions as associated with curiosity. CONCLUSIONS: Curiosity may be a critical precursor to smoking. Future research on smoking initiation should examine curiosity and investigate factors influencing it. PMID- 15833575 TI - A confirmatory factor analysis of the Fagerstrom Test for Nicotine Dependence. AB - This study used confirmatory factor analysis (CFA) to compare the relative fit of the following competing measurement models of the Fagerstrom Test for Nicotine Dependence (FTND): a single factor; two correlated factors (morning smoking and daytime smoking); and two correlated factors with one-item cross-loading. A single factor five-item version and a single factor four-item version of the FTND were also tested with the knowledge that these were not nested within the original six-item model. The subjects consisted of 231 smokers admitted for pre surgical assessment at a large urban teaching hospital. The results confirm the findings of previously published exploratory factor analyses (EFA) suggesting that the items of the FTND are best modeled as two correlated factors with a cross-loading. Given the origin of the FTND items as measures of physical dependence, the degree to which it has undergone largely data-driven modifications, and the confirmed complex structure of the items, it is recommended that researchers re-examine the extent to which the scale adequately represents current conceptualizations of nicotine dependence. PMID- 15833576 TI - Differences between Latino and non-Latino White smokers in cognitive and behavioral characteristics relevant to smoking cessation. AB - Adult smokers were recruited during routine health care visits at primary care clinics located in three urban hospitals and were given a brief intervention and nicotine replacement therapy. Analyses compared bicultural (BC: n=60) or less acculturated (LA: n=138) Latinos and non-Latino White (NL: n=417) participants. Both Latino groups were significantly different from NL subjects in smoking rate and nicotine dependence. However, BC and NL subjects differed significantly from LA subjects in perceived benefits of quitting, perceived risk from smoking, and negative affect smoking. LA subjects had higher cessation rates than either BC or NL groups. Regression analyses showed that nicotine dependence and confidence in quitting predicted cessation at month 6, and acculturation appeared to moderate the relationship between smoking cessation and both confidence in quitting and nicotine dependence. These results provide support for the viability of brief interventions for smoking provided through health care delivery systems. Results also suggest that characteristics previously shown to be predictive of successful cessation in mixed or non-Latino populations may not be equally predictive of cessation across members of diverse populations. PMID- 15833577 TI - Tobacco use, secondhand smoke exposure and their related knowledge, attitudes and behaviors among Asian Americans. AB - The present study examined tobacco use, secondhand smoke exposure and related knowledge, attitudes and behaviors among Asian Americans in the Delaware Valley of Pennsylvania and New Jersey, and the relationship between acculturation and smoking, social influence patterns on smoking, and stages of change of smoking among Asian subgroups. Study sample was 1174 Chinese, Koreans, Vietnamese, Cambodians, and other Asians. Findings revealed mean age of initiation to be 18.3, 40% ever and 30% current users. Significant differences were reflected in smoking by gender, ethnicity, educational level, marital and employment status. While knowledge and attitudes about smoking and secondhand smoke were associated with these variables, ethnic pride and smoking status played significant roles. Fathers and brothers had greater social influence on young male smoking behavior; smoking friends had influence on both genders. Stages of change of smoking and acculturation impact on smoking varied with gender, age, and time living in the U.S. Findings provide comprehensive insights into tobacco use and related KAB among Asian Americans that reflect the need for developing culturally appropriate programs for this underserved population. PMID- 15833578 TI - Mecamylamine moderates cue-induced emotional responses in smokers. AB - The nicotinic antagonist, mecamylamine, has been shown to reduce cue-elicited cocaine craving and to aid in smoking cessation. In a within-subjects design, 16 dependent smokers received mecamylamine (10 mg) or placebo capsules on two different days. Subjects imagined smoking urge and non-urge scenarios after smoking their usual brand vs. denicotinized cigarettes. Smoking usual-brand cigarettes produced greater positive effects and mecamylamine blocked heart rate (HR) boost and cigarette sensory impact. Mecamylamine also resulted in greater craving and less calmness, regardless of cigarette smoked. Urge script imagination in the mecamylamine+denicotinized condition resulted in calmness similar to usual-brand conditions and higher than the placebo+denicotinized condition. A similar trend was observed for negative affect. These results suggest that mecamylamine can moderate smoking cue-induced emotional responses in smokers. PMID- 15833579 TI - Determinants of positive and negative consequences of alcohol consumption in college students: alcohol use, gender, and psychological characteristics. AB - To examine the influence of alcohol consumption, gender, and psychological risk and protective factors on college students' experiences of negative and positive consequences, the present study of 181 students assessed frequency and quantity of alcohol consumption, negative and positive consequences of alcohol use, positive alcohol expectancies, constructive thinking, and positive and negative affect. Results indicated that men and women differed in their experience of some consequences and that while alcohol consumption was generally more strongly related to consequences for women than for men, it was unrelated to most consequences. Further, when controlling for alcohol consumption, positive alcohol expectancies and negative affect were positively related to experiencing positive and negative consequences while constructive thinking was related to fewer positive and fewer negative consequences. Results indicate that consequences are much more strongly related to psychological risk and protective factors than to alcohol consumption. The article concludes with a discussion of implications for intervention efforts. PMID- 15833580 TI - Acceptability of a computerized alcohol screening and advice routine in an emergency department setting--a patient perspective. AB - Disseminating screening and brief alcohol intervention in routine emergency department (ED) care seems to be a demanding challenge due to the complexity of the emergency setting. Further empirical realistic intervention models need to be developed that are appropriate for staff and patients before such models are tested in large scale controlled trials. This study explores patients' acceptance of a computerized opportunistic alcohol screening, followed by a personalized printout as feedback, at an emergency department performed by ordinary emergency department staff. In all, 44 patients were interviewed after performing the computerized screening test. The majority of patients were positive about answering the questions about alcohol habits, and 95% found it easy to use the touch screen and understand the instructions, even those with little computer experience. Regarding the personalized printout of advice, 76% were positive to this kind of feedback and 93% said they would read the printout. Most patients, 74%, preferred the computerized printout instead of a more personalized feedback from staff members. Whether the computerized advice is sufficient for a change in alcohol habits among excessive drinkers is still to be shown in controlled trials, but the concept could be one means of making preventive measures more feasible in a real-world ED setting. This might provide a basis for dissemination and integration of more extensive interventions in the long term. PMID- 15833581 TI - Quitting-related beliefs, intentions, and motivations of older smokers in four countries: findings from the International Tobacco Control Policy Evaluation Survey. AB - Older smokers represent an important subgroup that has been shown to benefit considerably from quitting smoking. However, to date little is known about relevant beliefs, intentions, and motivations. This study examined factors associated with older smokers' (aged 60 years and above) intention to quit smoking using data gathered via the International Tobacco Control Policy Evaluation Survey (ITCPES), a random digit dialed telephone survey of over 9000 adult smokers from United Kingdom, United States, Canada, and Australia. Having smoked for a long time and having survived, it was hypothesized that older smokers would perceive themselves as being less vulnerable to the harm of smoking (self-exempting beliefs); be less concerned about the health effects of smoking; be less confident about being able to quit successfully (self-efficacy); not perceive any health benefit of quitting, and hence be less willing to want to quit. Controlling for possible confounders, the hypotheses were all confirmed. Further analysis into reported considerations for quitting revealed that price of cigarettes, health professional advice, cheap quitting medication, and information on health risks were important predictors of quitting intention, with cigarette price and cheap medication also associated with recent quit attempts. Together, these findings have important implications for developing strategies for encouraging older smokers to give up smoking. PMID- 15833582 TI - Nonmedical use of prescription opioids among U.S. college students: prevalence and correlates from a national survey. AB - OBJECTIVES: This study examined the prevalence rates and correlates of nonmedical use of prescription opioid analgesics among U.S. college students in terms of student and college characteristics. METHODS: This study analyzed data from a nationally representative sample of 10,904 randomly selected students attending 119 four-year colleges in 2001. RESULTS: The lifetime prevalence of nonmedical prescription opioid use was 12% and the past year prevalence was 7%. Approximately one in every four colleges had a prevalence of 10% or higher for past year nonmedical use of prescription opioids. Multivariate regression analyses indicated nonmedical use was more likely to occur among college students who were white, residents of fraternity and sorority houses, attended more competitive colleges, earned lower grade point averages, and reported higher rates of substance use and other risky behaviors. CONCLUSIONS: This study provides evidence that the nonmedical use of prescription opioids represents a problem on college campuses. These findings have important implications for developing prevention efforts and therapeutic strategies aimed at reducing the nonmedical use of prescription opioid analgesics among college students while not hindering necessary medication management for pain. PMID- 15833583 TI - Fluoxetine in adolescents with comorbid major depression and an alcohol use disorder: a 3-year follow-up study. AB - The goal of this 3-year follow-up evaluation was to determine whether the decreases in drinking and in depressive symptoms that were noted during our acute phase study with fluoxetine in comorbid adolescents persisted at a 3-year follow up evaluation. At the 3-year follow-up evaluation, the group continued to demonstrate significantly fewer DSM criteria for an AUD and fewer BDI depressive symptoms and also consumed fewer standard drinks than they had demonstrated at the baseline of the acute phase study. However, 7 of the 10 participants demonstrated MDD at the 3-year follow-up assessment, and 4 demonstrated an AUD. The presence of a MDD was significantly correlated with the presence of an AUD at both the 1-year and the 3-year follow-up assessments. Four of the participants restarted SSRI medications during the follow-up period. Half of the subjects graduated from college during the 3-year assessment period, despite their residual depressive symptoms and drinking. We conclude that the long-term therapeutic effects of an acute phase trial of fluoxetine plus psychotherapy slowly decrease but did not disappear when fluoxetine is discontinued shortly after the acute phase trial. The high rate of MDD at follow-up suggests that longer term antidepressant medication treatment may be needed for at least some comorbid adolescents. PMID- 15833584 TI - Performance of heavy marijuana-smoking adolescents on a laboratory measure of motivation. AB - Marijuana smoking produces effects that may persist for hours or days beyond the period of acute intoxication. Despite evidence that adolescence represents a period of heightened exposure to marijuana, little research exists regarding possible impairment in adolescents who smoke marijuana regularly, and none exists regarding basic behavioral processes. In the present study, adolescents who smoked marijuana on a regular basis (near daily) were compared to a control group of adolescents on a two-option experimental task designed to measure motivation. The contingencies were arranged such that one option (work), which required systematically increasing response output, initially produced greater rates of monetary reinforcement than an alternative option (non-work) that required no response output to earn money. Switching to the non-work option was interpreted as a measure of reduced motivation. Significant differences were found between the groups: the marijuana-smoking participants switched earlier to the non-work option, and derived a greater percentage of their earnings from the non-work option. These differences existed when controlling for differences in cognitive aptitude, gender, and the presence of conduct disorder. A significant correlation between cannabinoid levels and percent of earnings derived from the non-work option suggests that these effects could be associated with the presence of cannabinoids in the marijuana-smoking group. PMID- 15833585 TI - Relations between cannabis use and dependence, motives for cannabis use and anxious, depressive and borderline symptomatology. AB - This study investigated the relations between anxious, depressive and borderline symptomatology, motivations for cannabis use, and cannabis use and dependence among 212 adolescents and young adults, 114 of whom were cannabis users. Motives for cannabis use were assessed using the Marijuana Motives Measure (Simons, J., Correia, C. J., Carey, K. B., & Borsari, B. E. (1998). Validating a Five-Factor Motives Measure: Relations with use, problems and alcohol motives. Journal of Counseling Psychology, 45, 265-273.). In three sets of regression analyses, motives, cannabis use frequency, and cannabis dependence served as criterion variables. First, when motives were regressed on psychopathological measures, borderline symptomatology predicted expansion motives in both boys and girls. Second, when frequency of use was regressed on motives and psychopathological measures, enhancement motives were the only significant predictor among boys and expansion motives were the only significant predictor among girls. Finally, when cannabis dependence was regressed on motives and psychopathological measures, borderline symptomatology was the only significant predictor in boys and expansion motives were the only significant predictor in girls. This study suggests the importance of motives and borderline symptomatology in the understanding of cannabis use and dependence among adolescents and young adults. PMID- 15833586 TI - Validity of self-reports and reliability of spousal proxy reports on the smoking behavior of Chinese parents with young children. AB - This study aims to examine the validity of self-reports and reliability of spousal proxy reports of smoking behavior among Chinese parents with young children. Agreement of self-reported smoking status with expired CO concentration among parents in the intake interview was significant with kappa at 0.70. Agreement of the mothers' and fathers' proxy reports on their spouses' smoking status and cigarette consumption by ICC were 1.0 and 0.58, and 0.82 and 0.33, respectively. Self-report and proxy reports of smoking status are considered valid and reliable to assess smoking behavior of parents with young children in Hong Kong. PMID- 15833587 TI - A gender specific psychometric analysis of the early trauma inventory short form in cocaine dependent adults. AB - This study evaluated the gender specific psychometric properties of the Early Trauma Inventory-Short Form (ETI-SF) in a clinical sample of cocaine dependent men (N=58) and women (N=34). Participants were administered the ETI-SF, the Childhood Trauma Questionnaire Short Form (CTQ-SF), and the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I). Tests of internal consistency, convergent validity, and predictive validity were conducted separately by gender. Findings indicated that the ETI-SF demonstrated good internal consistency; Cronbach's alpha ranged from 0.73 to 0.80 for men and from 0.70 to 0.77 for women. The measure also demonstrated good convergence with the CTQ-SF in both sexes, indicating that reports of child maltreatment are consistent across interview and self-report measures. Further, predictive validity was demonstrated by the ability of various ETI-SF scales to predict the co-occurrence of psychiatric disorders commonly associated with early trauma. These included lifetime diagnosis of PTSD in men and women, the lifetime diagnosis of major depressive disorder in men, and the lifetime diagnoses of alcohol use disorders in women. The findings support the utility of the ETI-SF as a clinical research tool to obtain data on specific types of early trauma in drug abusing samples. PMID- 15833588 TI - Correlation between self-reported smoking status and serum cotinine during pregnancy. AB - Maternal smoking is associated with adverse perinatal outcomes. Because of concerns of underreporting, investigators routinely perform biochemical testing to confirm smoking status, such as serum cotinine, a major metabolite of nicotine, adding an increased cost to examine compliance. The objectives of this study were to determine the sensitivity and specificity of self-reported smoking with serum cotinine as the gold standard and to determine the correlation between self-reported smoking in cigarettes per day and serum cotinine levels. In this cross-sectional study, we surveyed women during the first trimester of pregnancy on their tobacco exposure. A total of 40 women reported that they were smokers, and 40 were nonsmokers, 1 of whom had quit 5 days prior. The mean (+/-S.D.) serum cotinine value among smokers was 155 (+/-122) ng/l, vs. 1 (+/-6) ng/l in nonsmokers, p<0.001. The sensitivity of self-reported smoking status was 97.6%, and the specificity was 100%. Comparing the reported number of cigarettes smoked and the serum cotinine level, the Spearman correlation coefficient was 0.92 (p=0.015) overall and 0.67 (p=0.088) for the subgroup of smokers. This study demonstrates that self-reported smoking exposure during pregnancy is highly accurate. The high correlation coefficient suggests that this is a robust surrogate for cotinine levels. PMID- 15833589 TI - Effects of cortisol and cocaine on plasma prolactin and growth hormone levels in cocaine-dependent volunteers. AB - In rodents, corticosterone (cortisol in humans) facilitates cocaine self administration purportedly via enhancement of dopaminergic activity in the brain. This study sought to assess central dopaminergic effects of cortisol in humans and to compare them to those of cocaine. Twelve cocaine-dependent individuals received an intravenous bolus of cortisol (0.5 and 0.2 mg/kg; n=6 for each dose) and cocaine (0.2 mg/kg) in a double-blind randomized placebo-controlled and counterbalanced fashion. Their plasma was assayed over the next 120 min for prolactin and growth hormone (GH), which are two neuroendocrine indices of dopaminergic function. Cortisol injections produced significant increases in GH, while cocaine resulted in significant decreases in prolactin. Placebo administration was associated with gradual declines in prolactin, but the levels at the 90- and 120-min time points were significantly lower after cocaine than after placebo infusion. These different neuroendocrine response profiles point to important differences between dopaminergic effects of cortisol and cocaine. PMID- 15833590 TI - Age-related effects of IGF-1 on the NMDA-, GH- and IGF-1-receptor mRNA transcripts in the rat hippocampus. AB - Insulin-like growth factor-1 (IGF-1) and growth hormone (GH) have been suggested to promote memory and cognitive capabilities. In a recent publication we observed that GH increase the proportion of the NR2B subunit mRNA transcript of the NMDA receptor in rat hippocampus. NR2B has been suggested to be essential for spatial learning and long-term potentiation (LTP). This effect of GH might be IGF-1 mediated or a result of a co-ordination with IGF-1. To test this hypothesis further, we examined the effects of 10 daily s.c. injections of IGF-1 on NMDA receptor subunits (NR1, NR2A, and NR2B), GH receptor (GHR), GH binding protein (GHBP) and type 1 IGF receptor (IGF-1R) gene transcripts in the hippocampus. The NR2B subunit mRNA increased in young (11 weeks) but not in older (14-16 months) rats and the expression of the NR2A mRNA was decreased in both groups. The ratio of NR2B to NR2A is suggested to mirror the potential for synaptic plasticity. In both age groups, IGF-1 treatment resulted in a significant increase of this ratio at transcription level. The GHR mRNA increased in young rats, mimicking the effect of GH, while the IGF-1R mRNA was decreased in the older group of rats after IGF-1 treatment. These results suggest that IGF-1 in many aspects may mediate the actions earlier shown for GH. PMID- 15833591 TI - Increase of phosphorylation of calcium/calmodulin-dependent protein kinase-II in several brain regions by substance P administered intrathecally in mice. AB - In the present study, we investigated the role of phosphorylated calcium/calmodulin-dependent protein kinase-II (pCaMK-II) in nociceptive processing at the spinal and supraspinal levels in the substance P (SP)-induced mouse pain model. In the immunoblot assay, intrathecal (i.t.) injection with SP increased the pCaMK-II level in the spinal cord, and an immunohistochemical study showed that the increase of pCaMK-II immunoreactivity mainly occurred in the laminae I and II areas of the spinal dorsal horn. At the supraspinal level, pCaMK II was increased in the hippocampus and hypothalamus by i.t. SP injection, and an increase of pCaMK-II immunoreactivity mainly occurred in the pyramidal cells and the stratum lucidum/radiatum layer of the CA3 region of hippocampus and paraventricular nucleus of the hypothalamus. Moreover, pCaMK-II immunoreactivity in the locus coelureus of the brain stem was also increased. The nociceptive behavior induced by SP administered either i.t. or intracerebroventricularly (i.c.v.) was attenuated by KN-93 (a CaMK-II inhibitor). Our results suggest that pCaMK-II located at both spinal cord and supraspinal levels is an important regulator during the nociceptive processes induced by SP administered i.t. PMID- 15833592 TI - Human cortical responses during one-bit delayed-response tasks: an fMRI study. AB - Neuroimaging study of cognition across aging requires simple tasks ensuring: (i) high rate of correct performances in neurophysiological settings; and (ii) significant modulation of cortical activity. As a preliminary step, the present functional magnetic resonance imaging (fMRI) study tested the hypothesis that very simple delayed-response tasks fit these requirements in normal young adults. The short-term memory (STM) variant included a sequence of cue stimulus (two vertical bars), delay period (blank screen for only 5s), go stimulus, and motor response compatible with the taller vertical bar. Noteworthy, the retention (only one bit) could be based on visuo-spatial, phonological, and somatomotor coding. In the control variant (no STM, NSTM), the cue stimulus was present during the delay period. Results showed high rate of correct performances in both tasks (about 95%). Compared to the NSTM task (delay period), the STM task enhanced cortical responses in bilateral dorsolateral prefrontal (Brodmann area 8-9 (BA 8 9)), lateral premotor (BA 6L), medial premotor (BA 6M), inferior parietal (BA 40), and superior parietal (BA 7) areas. In the STM task, cortical responses were stronger in right than left BA 8-9 and BA 6L. These results indicate that, in normal young adults, a simple STM variant of delayed-response tasks (one bit to be retained) is correctly performed and enhances bilateral fronto-parietal responses. Therefore, it may be used for future cognitive neuroimaging studies on aging. PMID- 15833593 TI - Cobalt complex with GnRH stimulates the LH release and PKA signaling pathway in pig anterior pituitary cells in vitro. AB - Metal complexes with GnRH were shown to interact with GnRH receptors in pituitary cells. In the present study we examined the effects of GnRH and its cobalt complex form (Co-GnRH) on LH secretion and generation of second messengers, namely inositol phosphates (IPs) and cAMP, in porcine pituitary cells in vitro. The cells were obtained from gilt pituitary at the pre-ovulatory phase of estrous cycle and cultured for 72 h before challenge with GnRH or Co-GnRH. Both substances induced a significant increase in LH release that was detectable after 60 min (P<0.05) of treatment, with the Co-GnRH complex being more efficient than GnRH at 180 min (P<0.01). GnRH and Co-GnRH were equally effective at 10(-8)M (P<0.01), however, at the lowest (10(-9)M) as well as the highest (10(-7)M) concentrations tested, Co-GnRH was more potent than its native counterpart (P<0.01). Interestingly, Co-GnRH revealed twice more efficient than GnRH at stimulating cAMP production, an effect which was detectable in cells after 1h incubation (P<0.001). In contrast, while native GnRH induced a rapid increase (P<0.05) in IPs no such effect of Co-GnRH was observed. These data demonstrate that Co-GnRH and GnRH differentially effect on the signaling pathway in porcine gonadotropes and suggest that in these cells, the releasing action of Co-GnRH results from the mediation via the cAMP/protein kinase A second messenger system. PMID- 15833594 TI - M2/M4 muscarinic receptor binding in the anterior cingulate cortex in schizophrenia and mood disorders. AB - We have previously shown a decrease in [(3)H]pirenzepine binding to M1/M4 muscarinic receptors in the anterior cingulate cortex in schizophrenia but not in major depression or bipolar disorder. The present study aimed to extend these findings by examining the binding of [(3)H]AF-DX 384 to M2/M4 receptors in the same cohort of subjects. Using quantitative autoradiography we measured [(3)H]AF DX 384 binding in the anterior cingulate cortex of 15 schizophrenia, 15 bipolar, 15 major depression and 15 control cases. Post-mortem tissue was obtained from the Stanley Foundation Brain Bank. [(3)H]AF-DX 384 binding had a homogenous distribution amongst the layers of the anterior cingulate cortex, was higher in males than in females and declined with prolonged storage of tissue. An inverse correlation between [(3)H]AF-DX384 binding and age of onset of the disease was observed in the schizophrenia group suggesting that the earlier the age at onset the higher the binding was. In the depression group, there was a significant effect of gender on [(3)H]AF-DX 384 binding with females having lower binding in comparison to males. In the bipolar group, there was a significant inverse correlation between antipsychotic medication and [(3)H]AF-DX 384 binding, suggesting that the higher the dose of medication the lower the binding was. No differences in [(3)H]AF-DX 384 binding were seen between the four groups. The present results provide no evidence of M2/M4 receptor alterations in the anterior cingulate cortex in schizophrenia and affective disorders and extend the body of evidence implicating cortical M1 but not M2 involvement in the pathology and pharmacotherapy of schizophrenia. PMID- 15833595 TI - Occurrence of neuronal inclusions combined with increased nigral expression of alpha-synuclein within dopaminergic neurons following treatment with amphetamine derivatives in mice. AB - In recent years several clinical and research findings have demonstrated the involvement of the presynaptic protein alpha-synuclein in a variety of neurodegenerative disorders which are known as synucleinopathies. Although the function of this protein in the physiology of the cell remains unknown, it is evident that both genetic alterations or a mere overexpression of the native molecule produces a degeneration of nigral dopamine-containing neurons leading to movement disorders, as demonstrated in inherited Parkinson's disease. In the present study, we investigated whether widely abused drugs such as methamphetamine and methylenedioxymethamphetamine (ecstasy), which are known to damage the nigrostriatal dopamine pathway of mice, increase the expression of alpha-synuclein within dopamine neurons of the substantia nigra pars compacta. The results of this study demonstrate that nigrostriatal dopamine denervation and occurrence of intracellular inclusions in nigral neurons produced by amphetamine derivatives are related to increased expression of alpha-synuclein within dopamine neurons of the substantia nigra. This lends substance to the hypothesis that increased amounts of native alpha-synuclein may be per se a detrimental factor for the dopamine neurons. PMID- 15833596 TI - Caffeine induces differential cross tolerance to the amphetamine-like discriminative stimulus effects of dopaminergic agonists. AB - The purpose of this study was to determine if caffeine induces cross tolerance to the amphetamine-like discriminative stimulus effects of dopaminergic drugs that act through distinct mechanisms (e.g., release, uptake inhibition, direct activation of dopamine D(1)- or D(2)-family receptors). Rats were trained to discriminate 1.0 mg/kg d-amphetamine from saline in a two-choice discrete-trial procedure. Stimulus-generalization curves were generated by cumulative dosing for d-amphetamine (0.1-1.0 mg/kg), methylphenidate (0.3-5.6 mg/kg), SKF 81297 (0.3 3.0 mg/kg), and R-(-)-propylnorapomorphine (NPA; 0.001-1.78 mg/kg), as well as for caffeine (3.0-56 mg/kg); curves were re-determined after twice daily injections of caffeine (30 mg/kg) for 3.5 days. The rats generalized dose dependently to the four dopaminergic drugs, but only to a limited extent to caffeine. Twice daily injections of caffeine induced significant cross tolerance (i.e., increased ED(50)) to the amphetamine-like discriminative effects of methylphenidate and SKF 81297, attenuated non-significantly the effects of NPA, and did not alter the effects of amphetamine. Thus, caffeine produces differential cross tolerance to the amphetamine-like discriminative effects of dopaminergic drugs, a phenomenon in which the dopamine D(1) receptor appears to have an important role. PMID- 15833597 TI - Fos expression variances in mouse hypothalamus upon physical and osmotic stimuli: co-staining with vasopressin, oxytocin, and tyrosine hydroxylase. AB - Fos expression in the hypothalamus and its quantification in vasopressinergic (AVP), oxytocinergic (OXY) and tyrosine hydroxylase (TH) immunoreactive cells in the hypothalamic paraventricular (PVN), supraoptic (SON), suprachiasmatic (SCh), and arcuate (Arc) nuclei was performed in response to physiologically two different, i.e. osmotic (i.p. hypertonic saline, HS) and immobilization (IMO), stimuli in mouse using a dual Fos-neuropeptide immunohistochemistry. Both 60 min of HS and 120 min of IMO evoked Fos induction in many hypothalamic structures, whereas, HS evoked more extensive Fos labeling than IMO in the SON, ventromedial (VMN) and dorsomedial (NDM) hypothalamic nuclei and the retrochiasmatic area (RCh). Other hypothalamic structures including the anterior hypothalamic area (AHA), the latero-anterior hypothalamic nucleus (LA), the Arc, the perifornical nucleus (PeF), and the lateral hypothalamic area (LH) showed similar Fos incidence after both HS and IMO. However, after both stimuli explicitly most extensive Fos expression was observed in the PVN. In addition, in the PVN substantially more Fos-AVP (62-67% versus 10-15%) and Fos-OXY (38-45% versus 4 8%) perikarya were observed after HS than IMO, respectively. Incidence of TH immunoreactive Fos labeled cells in the PVN was also more frequent after HS. In the SON, HS activated more than 50% of AVP and OXY neurons while IMO less than 4%. The number of TH activated neurons in Arc was also higher after HS (11%) than IMO (4%). Lowest number of colocalizations was revealed in the SCh where both HS and IMO activated around 2% of AVP neurons. The present data demonstrate that both HS and IMO are powerful stimuli for the majority of hypothalamic structures displaying considerable topographic similarity in Fos expression suggesting their multifunctional involvement. The quantity and phenotypic differences of activated hypothalamic neurons may speak out for functional dissimilarities in response to HS and IMO. PMID- 15833598 TI - Further analysis of the effects of immunotoxic lesions of the basal nucleus of Meynert reveals substantial impairment on visual discrimination learning in monkeys. AB - In this paper we undertake a combined analysis of several studies in which marmoset monkeys received immunotoxic lesions of the cortical cholinergic projections from the basal nucleus of Meynert (NBM) bilaterally and/or in combination with immunotoxic lesions of other parts of the cholinergic system or ablations of the target inferotemporal neocortical area. Analysis of the mean learning scores across all visual discriminations learning tasks for each lesion combination revealed highly significant impairments where the NBM was lesioned bilaterally or where an NBM lesion in one hemisphere was crossed with an inferotemporal cortical ablation in the other hemisphere. This demonstrates that the cholinergic projection from the NBM to the major target area of neocortex involved in visual discrimination learning, i.e. the inferotemporal cortex, makes an important contribution to the perceptuo-mnemonic processes necessary for this type of learning. A new study demonstrates a significant effect of a subtotal bilateral cholinergic lesion confined to the NBM on a concurrent object-reward association task using black objects which is perceptually and mnemonically demanding. These results do not preclude the possibility that cholinergic projections from the NBM to other parts of the neocortex make a contribution to other cortical functions which are not mnemonic. It is well established that lesions of the cholinergic projection from the diagonal band of Broca disrupts the mnemonic functions of the hippocampus. The results described here suggest that degeneration of the cholinergic projections in Alzheimer's disease and other dementias will contribute to the loss of those mnemonic functions which are dependent on the neocortex. PMID- 15833599 TI - Acute and repeated restraint stress influences cellular damage in rat hippocampal slices exposed to oxygen and glucose deprivation. AB - Several studies have shown that high corticosteroid hormone levels increase neuronal vulnerability. Here we evaluate the consequences of in vivo acute or repeated restraint stress on cellular viability in rat hippocampal slices suffering an in vitro model of ischemia. Cellular injury was quantified by measuring lactate dehydrogenase (LDH) and neuron-specific enolase released into the medium. Acute stress did not affect cellular death when oxygen and glucose deprivation (OGD) was applied both immediately or 24h after restraint. The exposure to OGD, followed by reoxygenation, resulted in increased LDH in the medium. Repeated stress potentiated the effect of OGD both, on LDH and neuron specific enolase released to the medium. There was no effect of repeated stress on the release of S100B, an astrocytic protein. Additionally, no effect of repeated stress was observed on glutamate uptake by the tissue. These results suggest that repeated stress increases the vulnerability of hippocampal cells to an in vitro model of ischemia, potentiating cellular damage, and that the cells damaged by the exposure to repeated stress+OGD are mostly neurons. The uptake of glutamate was not observed to participate in the mechanisms responsible for rendering the neurons more susceptible to ischemic damage after repeated stress. PMID- 15833600 TI - Systemic Interleukin-1beta stimulates the simultaneous release of norepinephrine in the paraventricular nucleus and the median eminence. AB - Interleukin-1beta (IL-1beta), a cytokine with pronounced central effects such as fever, anorexia, analgesia, etc., is also known to activate the hypothalamo pituitary-adrenal (HPA) axis. Corticotropin releasing hormone (CRH) neurons located in the hypothalamus are important for HPA activation. The cell bodies of CRH neurons are located in the paraventricular nucleus (PVN) and their terminals are present in the median eminence (ME). Although the catecholamines, norepinephrine (NE) and dopamine (DA) are believed to be crucial factors in the stimulation of CRH neurons, it is not clear if they affect the cell bodies or terminals of these neurons to cause HPA activation. This study was done to determine if IL-1beta affects NE and DA release at the level of CRH cell bodies or their terminals. Adult male Sprague-Dawley rats were implanted with two push pull cannulae, one in the PVN and another in the ME, and were subjected to push pull perfusion. They were treated either with 0, 1 or 5 microg of IL-1beta. Perfusates were collected for 2 h after treatment and analyzed for NE concentrations using HPLC-EC. NE levels in the control and low dose groups did not change significantly during the entire period of observation both in the PVN and ME. In contrast, treatment with 5 microg of IL-1beta produced a marked increase in NE release in the PVN at 20 and 40 min post-treatment. NE release in the ME increased from 10 to 140 min post-treatment. There were no significant changes in the release of DA from both these areas. These results indicate that IL-1beta increases NE levels both in the PVN and in the ME and this could be a possible mechanism by which it stimulates the HPA axis. PMID- 15833601 TI - The seventh international conference on magnetic resonance applications to porous media. PMID- 15833602 TI - Magnetic resonance for fluids in porous media at the University of Bologna. AB - The magnetic resonance in porous media (MRPM) community is now a vast community of scientists from all over the world who recognize magnetic resonance as an instrument of choice for the characterization of pore space and of the distribution, diffusion and flow of fluids inside a vast range of different materials. The MRPM conferences are the occasions in which this community gets together, compares notes and grows. The scene was different in 1990, when this series of conferences was promoted at Bologna. I will go briefly over the history of these events, showing the role played by the University of Bologna and in particular by the intuition, ingenuity and passion of Giulio Cesare Borgia. The MRPM work at Bologna began in the mid-1980s. New correlations were found among parameters from NMR relaxation measurements and oil field parameters such as porosity, permeability to fluid flow, irreducible water saturation, residual oil saturation and pore-system surface-to-volume ratio, and fast algorithms were developed to give the different NMR parameters. Interest in valid interpretation of data led to extensive work also on the inversion of multiexponential relaxation data and the effects of inhomogeneous fields from susceptibility differences on distributions of relaxation times. In the last few years, extensive developments were made of combined magnetic resonance imaging and relaxation measurements in different fields. PMID- 15833603 TI - Some perspectives on dispersion and the use of ensemble-averaged PGSE NMR. AB - Magnetic resonance methodology has made a significant impact in helping us understand the physics of porous media. Among an important class of experiments is that set of techniques designed to measure fluid dispersion. This paper provides some background on some of the underlying physics of dispersion, and outlines some of the NMR approaches that have proven successful. The local and nonlocal dispersion tensors are described and the prospects for future NMR advances considered. PMID- 15833604 TI - Long-range diffusion in beds of nanoporous particles: pitfalls and potentials. AB - Owing to the recent progress in the area of hardware and software of the pulsed field gradient NMR technique, molecular transport in real-life zeolite systems, such as zeolite beds and particles of formulated fluid catalytic cracking (FCC) catalysts, can be investigated in detail. These studies have revealed a number of important features of molecular transport in zeolites, which are reviewed in the present paper. In particular, the anomalous character of intracrystalline diffusion in MFI-type zeolites, dependence of the tortuosity factor in zeolite beds on diffusion regime and the role of various modes of diffusion in transport limitations arising for catalytic reactions in FCC catalysts will be discussed. PMID- 15833605 TI - Confined dynamics, forms and transitions in colloidal systems: from clay to DNA. AB - Colloidal suspensions are a classic example of confining systems developing large specific surfaces, presenting a rich variety of shapes and exhibiting complex organization on a length scale ranging from 1 nm to several micrometers. Two distinct confined dynamics are generally considered in such systems: (1) the embedded fluid dynamics entrapped in the pore network with two main contributions, surface interaction and long-range connectivity, and (2) the dynamics of the host matrix, associated with a time evolution of the interfacial geometry. This last contribution is particularly important during dynamic and structural transitions of colloidal suspensions such as jamming, glass transition, phase separations and flocculation. It is generally believed that the characteristic time scale needed to describe colloidal movement and interfacial geometrical reorganization is much slower than the dynamics of the embedded fluid (except in the trivial situation where the fluid molecule is irreversibly adsorbed to a colloidal surface). Thus, few connections are made between these two distinct dynamics. In this presentation, we show how the slow and confined water dynamics at proximity of a colloidal surface provides an original way to probe colloidal shape and colloidal orientation dynamics. Two topics are presented. First of all, water field-cycling NMR relaxometry is used to probe the glass transition and the strong rotational slowing down of a colloidal system made of plate-like particles, a synthetic clay (laponite). Second, we analyze the case of long colloidal thin rods (either mineral or biologic such as DNA cylinders) dispersed in very diluted suspensions. At large distance and/or long time, these particles appear as a portion of a line. We discuss how the embedded fluid dynamics can be sensitive to this morphological crossover and may provide information about the particle shape. Some comparisons with recent experiments are presented. PMID- 15833606 TI - Hyperpolarization of 13C through order transfer from parahydrogen: a new contrast agent for MRI. AB - The order within proton pairs in organic molecules, resulting from hydrogenation with parahydrogen, can be transferred in great part to nearby carbon 13 spins through adequate field manipulations. The molecules with hyperpolarized 13C thus obtained can be used as new contrast agents of high efficiency in MRI. After a brief presentation of the hydrogenation process and apparatus, in relatively low magnetic field, we describe the procedure of order transfer to the 13C spins through a sudden drop from the initial field to zero field followed by an adiabatic remagnetization. The expected final polarizations in the absence of relaxation are given for several compounds. Finally, we show an example of MR images observed in vivo on animals as an illustration of the contrast capacity of this new method. PMID- 15833607 TI - Viscoelastic shear properties of in vivo breast lesions measured by MR elastography. AB - Elastography is a technique to assess the viscoelastic properties of tissue by measuring an acoustic wave propagating though the object. Here, the technique is applied in the course of standard MR mammography to 15 patients with different pathologies (six breast cancer cases, six fibroadenoma cases and three mastopathy cases). Low-frequency mechanical waves are coupled longitudinally into the tissue in order to obtain sufficient wave amplitude throughout the entire breast. This leads to the presence of a substantial fraction of compressional waves, which contribute to the total displacement field. It is shown theoretically that the correct evaluation of these contributions from the compressional wave is rather difficult due to the almost incompressible nature of tissue. To overcome this problem, it is proposed to apply the curl-operator to the measured displacement field in order to completely remove contributions from the compressional wave. Results from simulations and a breast phantom demonstrate the feasibility of the technique. The in vivo results show a good separation between breast cancer and benign fibroadenoma utilizing the shear modulus. Breast cancer appears on average 2.2 (P<.001) times stiffer. All breast cancer cases showed a good delineation to the surrounding breast tissue with an average elevation of a factor of 3.3 (P< 1.4 x 10(-6)). The results as obtained for the shear viscosity do not indicate to be useful for separating benign from malignant lesions. PMID- 15833608 TI - Nuclear magnetic resonance and spin relaxation in biological systems. AB - Proton nuclear spin-lattice relaxation in biological systems is generally distinguished from that in inorganic systems such as rocks by the presence of locally disordered macromolecular environments. Rapid exchange of readily observed labile small molecules among differently oriented macromolecular sites generally nearly averages the spectral anisotropies in the small molecule resonances. The biological tissue is generally distinguished from the inorganic matrix by the presence of a significant population of protons in the solid components that are well connected by dipolar spin couplings. Magnetic coupling between the solid and the liquid components generally dominates the magnetic field dependence of the spin-lattice relaxation rates observed in the small molecule components which is generally described by a power law in the Larmor frequency. Recent theory involving a modification of the spin-phonon class of relaxation mechanism provides a quantitative understanding of these data in terms of the dynamics of the chain molecules generally present in the solid spin systems, folded proteins for example. PMID- 15833609 TI - Field-cycled PEDRI imaging of free radicals with detection at 450 mT. AB - This paper describes the design, construction and use of a field-cycled proton electron double-resonance imaging (FC-PEDRI) system for the detection and imaging of free radicals. The unique feature of this imager is its use of a 450-mT detection magnetic field in order to achieve good image quality and sensitivity. The detection magnetic field is provided by a superconducting magnet, giving high stability and homogeneity. Field cycling is implemented by switching on and off the current in an internal, coaxial, resistive secondary magnet that partially cancels the superconducting magnet's field at the sample; the secondary magnet is actively shielded to avoid eddy currents. EPR irradiation takes place at approximately 5 mT, following which the field is switched to 450 mT in 40 ms for NMR signal detection. Full details of the imager's subsystems are given, and experiments to image the distribution of stable free radical contrast agents in phantoms and in anesthetized rats are described. PMID- 15833610 TI - Proton electron double resonance imaging of free radical distribution in environmental science applications--first results and perspectives. AB - In this contribution, we explore the potential of proton electron double resonance imaging (PEDRI) in environmental science (hydrogeological) applications. After a discussion of the hydrogeological motivation for studies of free radical transport in environmental matrices, we present results from first experiments that show the principal applicability of the PEDRI technique to sediment samples. Field-cycled (FC) relaxation time contrast is identified as a possible source of artifacts in samples in which strong concentration gradients of the free radical phase are present. Furthermore, an outlook is given on how PEDRI can help in observations of the local interplay among contaminants, water and nonaqueous liquid phases. PMID- 15833611 TI - Polymer chain dynamics under nanoscopic confinements. AB - It is shown that the confinement of polymer melts in nanopores leads to chain dynamics dramatically different from bulk behavior. This so-called corset effect occurs both above and below the critical molecular mass and induces the dynamic features predicted for reptation. A spinodal demixing technique was employed for the preparation of linear poly(ethylene oxide) (PEO) confined to nanoscopic strands that are in turn embedded in a quasi-solid and impenetrable methacrylate matrix. Both the molecular weight of the PEO and the mean diameter of the strands were varied to a certain degree. The chain dynamics of the PEO in the molten state was examined with the aid of field-gradient NMR diffusometry (time scale, 10(-2)-10(0) s) and field-cycling NMR relaxometry (time scale, 10(-9)-10(-4) s). The dominating mechanism for translational displacements probed in the nanoscopic strands by either technique is shown to be reptation. On the time scale of spin lattice relaxation time measurements, the frequency dependence signature of reptation (i.e., T1 approximately nu(3/4)) showed up in all samples. A "tube" diameter of only 0.6 nm was concluded to be effective on this time scale even when the strand diameter was larger than the radius of gyration of the PEO random coils. This corset effect is traced back to the lack of the local fluctuation capacity of the free volume in nanoscopic confinements. The confinement dimension is estimated at which the crossover from confined to bulk chain dynamics is expected. PMID- 15833612 TI - Advances of unilateral mobile NMR in nondestructive materials testing. AB - Unilateral mobile NMR employs portable instrumentation with sensors, which are applied to the object from one side. Based on the principles of well-logging NMR, a hand-held sensor, the NMR-MOUSE (MObile Universal Surface Explorer) has been developed for nondestructive materials testing. In the following, a number of new applications of unilateral NMR in materials science are reviewed. They are the state assessment of polyethylene pipes, the characterization of wood, the in situ evaluation of stone conservation treatment, high-resolution profiling of rubber tubes and 2-D imaging for defect analysis in rubber products. PMID- 15833613 TI - Study of gas-fluidization dynamics with laser-polarized 129Xe. AB - We report initial NMR studies of gas dynamics in a particle bed fluidized by laser-polarized xenon (129Xe) gas. We have made preliminary measurements of two important characteristics: gas exchange between the bubble and emulsion phases and the gas velocity distribution in the bed. We used T2* contrast to differentiate the bubble and emulsion phases by choosing solid particles with large magnetic susceptibility. Experimental tests demonstrated that this method was successful in eliminating 129Xe magnetization in the emulsion phase, which enabled us to observe the time dependence of the bubble magnetization. By employing the pulsed field gradient method, we also measured the gas velocity distribution within the bed. These results clearly show the onset of bubbling and can be used to deduce information about gas and particle motion in the fluidized bed. PMID- 15833614 TI - Concentration-dependent self-diffusion of adsorbates in mesoporous materials. AB - The pulsed-field gradient NMR method has been applied to study self-diffusion of liquids in mesoporous materials with different pore sizes and morphologies as a function of pore loading. It is found that the effective diffusivities of adsorbate molecules in mesopores at partial loadings are related to two mechanisms, the Knudsen diffusion through the gaseous phase in the pore space and the diffusion within the layer of molecules adsorbed on the pore walls. The relative contributions of these modes, which are determined by the details of the interphase equilibrium, change with variation of the pore loading, leading to a complex behavior of the effective self-diffusion coefficient. The impact of the pore size and the adsorbate-surface interaction on self-diffusion is elucidated. Possible reasons for an experimentally obtained hysteresis in the diffusivities measured on adsorption and desorption in mesopores are discussed. PMID- 15833615 TI - Modeling diffusion in white matter in the brain: a composite porous medium. AB - We model diffusion in white matter fascicles as a problem of diffusion in an array of identical thick-walled cylindrical tubes immersed in an outer medium and arranged periodically in a regular lattice. The diffusing molecules have different diffusion coefficients and concentrations (or densities) within the tubes' inner core, membrane, myelin sheath, and within the outer medium. For an impermeable myelin sheath, diffusing molecules within the inner core are completely restricted, while molecules in the outer medium are hindered due to the tortuosity of the array of impenetrable tubes. PMID- 15833616 TI - Functional MRI and NMR spectroscopy of an operating gas-liquid-solid catalytic reactor. AB - A dynamic in situ study of alpha-methylstyrene catalytic hydrogenation on a single catalyst pellet or in a granular bed is performed using 1H MRI and spatially resolved 1H NMR spectroscopy. Owing to reaction exothermicity, a reciprocating motion of the liquid front within the pellet accompanied by pellet temperature oscillations has been observed. Spatially resolved information on the reactant to product conversion within the catalyst bed has been obtained for a steady-state regime. Two-dimensional 27Al NMR images of alumina catalyst supports and other alumina-containing materials have been detected using moderate magnetic field gradients (80 G/cm) and a two-pulse spin-echo sequence. Temperature dependence of signal intensity and 27Al T1 time of alumina are considered as possible temperature sensors for NMR thermometry applications. PMID- 15833617 TI - Combining macroscopic and microscopic diffusion studies in zeolites using NMR techniques. AB - In this study the zero length column (ZLC) technique is extended to the case where the decay of the adsorbed phase concentration is observed directly by nuclear magnetic resonance (NMR). An adsorption-desorption apparatus compatible with a 400-MHz NMR spectrometer was developed. It operates with nitrogen or helium as the inert purge gas. The column of the adsorbent material is placed in the sensitive region of the superconducting magnet and the rf coil of the NMR spectrometer. The time scales of the adsorption and desorption processes depend on concentration, temperature and crystal shape and are found to be in the range of 1-10 min. From the desorption branch, the non-equilibrium ZLC-NMR measurements yield intracrystalline diffusion coefficients in the range of 10(-13) to 10(-11) m2/s for different alkanes in silicalite-1. These values are always found to be smaller than the values measured by pulsed field gradient NMR under equilibrium condition indicating that there must be additional transport resistance at the external surface of these silicalite-1 zeolite crystals. PMID- 15833619 TI - Contrast-enhanced dynamic MRI protocol with improved spatial and time resolution for in vivo microimaging of the mouse with a 1.5-T body scanner and a superconducting surface coil. AB - Magnetic resonance imaging (MRI) is well suited for small animal model investigations to study various human pathologies. However, the assessment of microscopic information requires a high-spatial resolution (HSR) leading to a critical problem of signal-to-noise ratio limitations in standard whole-body imager. As contrast mechanisms are field dependent, working at high field do not allow to derive MRI criteria that may apply to clinical settings done in standard whole-body systems. In this work, a contrast-enhanced dynamic MRI protocol with improved spatial and time resolution was used to perform in vivo tumor model imaging on the mouse at 1.5 T. The needed sensitivity is provided by the use of a 12-mm superconducting surface coil operating at 77 K. High quality in vivo images were obtained and revealed well-defined internal structures of the tumor. A 3-D HSR sequence with voxels of 59x59x300 microm3 encoded within 6.9 min and a 2-D sequence with subsecond acquisition time and isotropic in-plane resolution of 234 microm were used to analyze the contrast enhancement kinetics in tumoral structures at long and short time scales. This work is a first step to better characterize and differentiate the dynamic behavior of tumoral heterogeneities. PMID- 15833618 TI - Pulsed-field gradient nuclear magnetic resonance study of transport properties of fluid catalytic cracking catalysts. AB - Pulsed-field gradient nuclear magnetic resonance (PFG NMR) has been applied to study molecular diffusion in industrial fluid catalytic cracking (FCC) catalysts and in USY zeolite for a broad range of molecular displacements and temperatures. The results of this study have been used to elucidate the relevance of molecular transport on various displacements for the rate of molecular exchange between catalyst particles and their surroundings. It turned out that this rate, which may determine the overall rate and selectivity of FCC process, is primarily related to the diffusion mode associated with displacements larger than the size of zeolite crystals located in the particles but smaller than the size of the particles. This conclusion has been confirmed by comparative studies of the catalytic performance of different FCC catalysts. PMID- 15833620 TI - DTI of trabecular bone marrow. AB - The development of NMR diffusion imaging and diffusion tensor imaging (DTI) has offered the possibility of studying the porous structures beyond anatomical imaging. In fact, random molecular motions, within tissue components, probe tissue microstructures. Up to now, the DTI method was mainly used to investigate cerebral morphology and study white matter diseases. In this study, it has been applied to trabecular bone marrow analysis to obtain structural information on spongy bone tissue. Our first results show that DTI could represent an important tool in studying the microstructural architecture of the trabecular bone as well as the microarchitecture of porous media. PMID- 15833621 TI - Morphology and chain dynamics during collapse transition of PNIPAM gels studied by combined imaging, relaxometry and 129Xe spectroscopy techniques. AB - The temperature-induced shape transition of poly(N-isopropylacrylamide) gels of different cross-link densities was investigated by a combination of NMR techniques allowing the characterization of both the macroscopic collapse as well as the changes on a molecular scale related to the expulsion of water from the gel network. The proton-containing gel phase was visualized by swelling in heavy water, and the volume change was monitored by proton imaging for cross-link densities between 0.5% and 2.5%. Above the transition temperature of 35 degrees C, gel collapse led to a volume change of up to a factor of 17 for the gel of smallest cross-link density. Two spectral lines of 129Xe are found in the gel state and are assigned to the hydrophobic and hydrophilic parts of the network. In the collapsed state, the hydrophobic peak shows a strong shift while the hydrophilic peak disappears. A considerable shortening of both T1 and T2 of the gel protons upon collapse was found at a field of 4.7 T. At lower fields, the effect becomes more pronounced and qualitatively different dispersion behaviors between the swollen and the collapsed states are observed. PMID- 15833622 TI - Water content distribution in a polymer electrolyte membrane for advanced fuel cell system with liquid water supply. AB - To better understand the operation of a new fuel cell design, we used magnetic resonance imaging (MRI) to measure the water content distribution in a polymer electrolyte membrane under fuel cell operation with and without a supply of liquid water. The supply of liquid water to the membrane improved the cell performance by increasing the water content in the membrane and thus reducing the electrical resistance of the membrane. The study also showed that MRI is a promising method to investigate the distribution of water in the membrane of a fuel cell under operating conditions. PMID- 15833623 TI - Two-dimensional nuclear magnetic resonance petrophysics. AB - Two-dimensional nuclear magnetic resonance (2D NMR) opens a wide area for exploration in petrophysics and has significant impact to petroleum logging technology. When there are multiple fluids with different diffusion coefficients saturated in a porous medium, this information can be extracted and clearly delineated from CPMG measurements of such a system either using regular pulsing sequences or modified two window sequences. The 2D NMR plot with independent variables of T2 relaxation time and diffusion coefficient allows clear separation of oil and water signals in the rocks. This 2D concept can be extended to general studies of fluid-saturated porous media involving other combinations of two or more independent variables, such as chemical shift and T1/T2 relaxation time (reflecting pore size), proton population and diffusion contrast, etc. PMID- 15833624 TI - Centric scan SPRITE for spin density imaging of short relaxation time porous materials. AB - The single-point ramped imaging with T1 enhancement (SPRITE) imaging technique has proven to be a very robust and flexible method for the study of a wide range of systems with short signal lifetimes. As a pure phase encoding technique, SPRITE is largely immune to image distortions generated by susceptibility variations, chemical shift and paramagnetic impurities. In addition, it avoids the line width restrictions on resolution common to time-based sampling, frequency encoding methods. The standard SPRITE technique is however a longitudinal steady-state imaging method; the image intensity is related to the longitudinal steady state, which not only decreases the signal-to-noise ratio, but also introduces many parameters into the image signal equation. A centric scan strategy for SPRITE removes the longitudinal steady state from the image intensity equation and increases the inherent image intensity. Two centric scan SPRITE methods, that is, Spiral-SPRITE and Conical-SPRITE, with fast acquisition and greatly reduced gradient duty cycle, are outlined. Multiple free induction decay (FID) points may be acquired during SPRITE sampling for signal averaging to increase signal-to-noise ratio or for T2* and spin density mapping without an increase in acquisition time. Experimental results show that most porous sedimentary rock and concrete samples have a single exponential T2* decay due to susceptibility difference-induced field distortion. Inhomogeneous broadening thus dominates, which suggests that spin density imaging can be easily obtained by SPRITE. PMID- 15833625 TI - Interactions between chloride and cement-paste materials. AB - The durability of cement-based materials with respect to exterior aggressions is one of the current priorities in civil engineering. Depending on their use, the cement-based materials can be exposed to different types of aggressive environments. For instance, damages to concrete structures in contact with a saline environment (sea water on bridges, deicing salts on roads, etc.) are of utmost importance. Upon exposure to saline water, Cl- ions penetrate into the structures and subsequently lead to reinforcement corrosion. Chloride attack is often combined with other aggressive influences such as temperature (e.g., freezing) or the ingress of other ions (e.g., sulfates in sea water). We therefore aim to explore the effect of sodium chloride (NaCl) on the structural chemistry of cement paste. Existing studies about reinforcement corrosion by chloride have focused on the penetration of Cl- ions and the comparison between "free" ions (water-soluble ions) and bound ones. However, little is known about the fixation mechanisms, the localization of Cl in the cement matrix and the structural interaction between Cl and the silicate and aluminate hydrate phases present in cement paste. We present here results of a multinuclear nuclear magnetic resonance study on the fixation of chloride in the hydration products and the characterization of new phases potentially appearing due to chloride ingress. PMID- 15833626 TI - Salt crystallization as damage mechanism in porous building materials--a nuclear magnetic resonance study. AB - Salts can damage building materials by chemical reactions or crystallization, which is a serious threat to cultural heritage. In order to develop better conservation techniques, more knowledge of the crystallization processes is needed. In a porous material, the size of a salt crystal is limited by the sizes of the pores. It has been predicted that as a consequence, the solubility of a salt increases with decreasing pore size. This increase seems to be related to an increase of the stress generated by a crystal on the pore wall. It has been suggested that the resulting stress could become high enough to induce failure. We have studied the crystallization of salts in porous materials with well defined pore sizes. Samples were saturated at 40 degrees C with saturated Na2SO4 and Na2CO3 solutions. Next we have cooled the samples to 0 degrees C and waited for nucleation. After nucleation occurred, the solubility in the porous material was measured with nuclear magnetic resonance (NMR) as a function of the temperature. The measurements on Na2CO3 indeed show an increase in solubility with a decrease in pore size. For Na2SO4, we did not observe a pore size dependent solubility. However, we have to remark that these results show a metastable crystal phase. The results can be used to calculate the actual pressure exerted by the crystals onto the pore wall. PMID- 15833627 TI - The influence of superplasticizers on the first steps of tricalcium silicate hydration studied by NMR techniques. AB - The influence of superplasticizer sulfonated naphthalene formaldehyde (SNF) on the hydration process of tricalcium silicate (C3S) paste was investigated by (1)H nuclear magnetic resonance spin-spin and spin lattice relaxation times. The addition of SNF superplasticizer to C3S paste clearly affects the morphology and growth rates of the hydration products, mainly by increasing the dormant period length, which lasts for several hours more than in conventional C3S hydrated paste, while reducing the acceleration period length. The relaxation data indicated that a pronounced delay occurs in the C3S hardening when sulfonated polymers are added to the makeup water. For all the analyzed samples, prepared with a water-to-C3S ratio of 0.4, the decay of the echo magnetization has been fitted by adopting both a monoexponential and a biexponential relaxation model in order to evaluate the contributions from water in different regimes of hydration. PMID- 15833628 TI - NMR study of the vapor phase contribution to diffusion in partially filled silica glasses with nanometer and micrometer pores. AB - The contribution of the vapor phase to molecular diffusion in porous silica glasses with nanometer (Vycor) and micrometer (VitraPor#5) pores partially filled with water (polar) or cyclohexane (nonpolar) was investigated with the aid of field-gradient NMR diffusometry. Due to the vapor phase, the effective diffusion coefficient of cyclohexane filling micrometer pores (VitraPor#5) increased up to 10 times relative to the value in bulk liquid upon reduction of the pore space filling factor. On the other hand, the effective diffusion coefficient of water first decreases and then increases when the liquid content is reduced. The dependence of the effective diffusion coefficient on the pore filling factor is strongly related to the pore dimension. A general two-phase exchange model is presented that is well accounting for all experimental diffusion features. PMID- 15833629 TI - Hydrodynamics in two-phase flow within porous media. AB - Ultra-fast magnetic resonance imaging techniques are used to image liquid distribution in two and three dimensions during air-water co-current down flow through a fixed bed of cylindrical porous pellets of length and diameter 3 mm, packed within a 43 mm internal diameter column in both the trickle- and pulsing flow regimes. The data acquisition times used were 20 and 280 ms, giving 2-D and 3-D spatial resolutions of 1.4 mm x 2.8 mm and 3.75 mm x 3.75 mm x 1.87 mm, respectively. This work reports images of local pulsing events within the bed occurring during the trickle-to-pulse flow transition. The evolution of the local instabilities is studied as a function of increasing liquid velocity at constant gas velocity. PMID- 15833630 TI - NMR in rotating magnetic fields: magic-angle field spinning. AB - Magic-angle sample spinning is one of the cornerstones in high-resolution NMR of solid and semisolid materials. The technique enhances spectral resolution by averaging away rank 2 anisotropic spin interactions, thereby producing isotropic like spectra with resolved chemical shifts and scalar couplings. In principle, it should be possible to induce similar effects in a static sample if the direction of the magnetic field is varied (e.g., magic-angle rotation of the B0 field). Here we will review some recent experimental results that show progress toward this goal. Also, we will explore some alternative approaches that may enable the recovery of spectral resolution in cases where the field is rotating off the magic angle. Such a possibility could help mitigate the technical problems that render difficult the practical implementation of this method at moderately strong magnetic fields. PMID- 15833631 TI - Multiple modulation multiple echoes: a one-shot method. AB - This paper reviews a recently reported NMR method capable of determining the diffusion constant within milliseconds and without the need of multiple scans. The method can be used with static or pulsed magnetic field gradients. PMID- 15833632 TI - Fixed and pulsed gradient diffusion methods in low-field core analysis. AB - We review diffusion-weighted relaxation protocols for two-dimensional diffusion/relaxation time (D, T(2)) distributions and their application to fluid saturated sedimentary rocks at low fields typical of oil-well logging tools (< or = 2 MHz for 1H). Fixed field gradient (FFG) protocols may be implemented in logging tools and in the laboratory; there, pulsed field gradient (PFG) protocols are also available. In either category, direct or stimulated echoes may be used for the diffusion evolution periods. We compare the results of several variant FFG and PFG protocols obtained on liquids and two contrasting sedimentary rocks. For liquids and rocks of negligible internal gradients (g(int)), results are comparable, as expected, for all the studied protocols. For rocks of strong g(int), protocol-dependent artifacts are seen in the joint (D, T2) distributions, consistent with the effects of the internal fields. For laboratory petrophysics, the PFG methods offer several advantages: (a) significantly improved signal-to noise ratio and acquisition times for repetitions over many samples; (b) freedom from heteronuclear contamination when fluorinated liquids are used in core holders; and (c) a palette of variants--one comparable with the FFG--for the study of rocks of significant g(int). Given suitable hardware, both PFG and FFG methods can be implemented in the same bench-top apparatus, providing a versatile test bed for application in a petrophysical laboratory. PMID- 15833633 TI - An MRI-SPI and NMR relaxation study of drying-hydration coupling effect on microstructure of cement-based materials at early age. AB - Hydration and drying coupling effect monitoring with single point imaging profiles allowed us to evaluate "free" pore and "bounded" chemical water quantity. White cement pastes inverse Laplace analysis of T1 measurements shown original results with two components during setting. After hardening, we found three components. Single point imaging measures were also used to study the evolution of transition zone in repaired concrete. MRI results demonstrated its interest compared with destructive method for longitudinal study and phenomena kinetic monitoring. PMID- 15833634 TI - Dissolution of laser-polarized xenon in benzene. AB - The study of the dissolution of laser-polarized xenon in degassed deuterated benzene is reported. We show that the time evolution of the xenon signal implies that a transient convective process takes place. It is characterized by velocity encoding magnetic resonance measurements and MRI experiments. PMID- 15833635 TI - Continuous wave MRI diffusion study of water in bentonite clay. AB - Three-layered clay minerals such as montmorillonite (bentonite) exhibit very short transverse relaxation times. This is especially true for samples with relatively low water contents in the region of 20% to 30%, which is the water content typically used in environmental technology applications (e.g., as a mineral liner material for landfills). The diffusion of water in samples with such short transverse relaxation times can be measured with NMR by observing the moisture gradients or isotope tracer fronts propagating through appropriately prepared samples by means of continuous wave MRI. The first results from such studies on bentonite clays are presented in this paper. PMID- 15833636 TI - NMR investigation of gaseous SF6 confinement into EPDM rubber. AB - The confinement process of gaseous sulphurhexafluoride (SF6) in ethylene propylene-diene (EPDM) rubber was investigated by spectroscopic and spatially resolved NMR techniques. A strong elongation of T1 relaxation time of SF6 and a decrease of the diffusion coefficient were found. A possible explanation may be the strong restriction of molecular mobility due to interactions between SF6 and active centers of the EPDM. PMID- 15833637 TI - Imaging of water distribution in thermally fractured granites by SPRITE. AB - Water distribution in thermally fractured granite samples was visualized by using SPRITE sequences. Networks of intergranular fractures were observed in the coarse grained Inada granite after heating at 873 K or above. On the other hand, bright spots were observed in the fine-grained Okazaki granite, which may be due to pore water in feldspar grains. D2O diffusion into samples saturated with H2O was also observed by 2D-projected SPRITE imaging. PMID- 15833638 TI - Xenon NMR measurements of permeability and tortuosity in reservoir rocks. AB - In this work we present measurements of permeability, effective porosity and tortuosity on a variety of rock samples using NMR/MRI of thermal and laser polarized gas. Permeability and effective porosity are measured simultaneously using MRI to monitor the inflow of laser-polarized xenon into the rock core. Tortuosity is determined from measurements of the time-dependent diffusion coefficient using thermal xenon in sealed samples. The initial results from a limited number of rocks indicate inverse correlations between tortuosity and both effective porosity and permeability. Further studies to widen the number of types of rocks studied may eventually aid in explaining the poorly understood connection between permeability and tortuosity of rock cores. PMID- 15833639 TI - Network images of drainage channels in sea spray icing by MR microscopy. AB - An MR imaging system was developed to visualize brine drainage channels in sea spray icing. Brine pockets trapped in spray-ice matrices during ice growth are structural features of sea spray icing. Brine in the spray ice sample had drained out; therefore, using a suction pump, we filled the air gaps in the drainage channels with dodecane. In the experiments, 0.5-1.0 h was necessary to accumulate signals sufficient to obtain a 3-D micro-image; the image matrix comprised 128(3) voxels (each voxel was 200 microm per side). The MIP view showed that sea spray icing has a developed drainage-channel network structure. PMID- 15833640 TI - Measuring pore connectivity by pulsed field gradient diffusion editing with hydrocarbon gases. AB - Measurements of time-dependent diffusion are performed on a rock sample saturated first with water, then methane and finally ethane. The gases were selected because their increased diffusivities and relaxation times allow probing greater length scales than water and because of their practical relevance. The nuclear magnetic resonance measurements employed pulse field gradient diffusion editing pulse sequences, allowing analysis of D(t) as a function of relaxation time. Very different D(t) behaviors are observed for different relaxation times, including indications of connected pore networks at moderate relaxation times. PMID- 15833641 TI - MRI as a key tool for understanding and modeling the filtration kinetics of fibrous media. AB - Applying MRI techniques to low-density fibrous filter media provides us with unique information about the initial structure and deposited mass within the same filter sample. This now enables us to obtain the necessary link between structure and deposition for validation and further enhancement of modeling filtration kinetics. However, additional work is needed before achieving a realistic understanding of filtration kinetics. PMID- 15833642 TI - Correlation of porous and functional properties of food materials by NMR relaxometry and multivariate analysis. AB - The porous properties of food materials are known to determine important macroscopic parameters such as water-holding capacity and texture. In conventional approaches, understanding is built from a long process of establishing macrostructure-property relations in a rational manner. Only recently, multivariate approaches were introduced for the same purpose. The model systems used here are oil-in-water emulsions, stabilised by protein, and form complex structures, consisting of fat droplets dispersed in a porous protein phase. NMR time-domain decay curves were recorded for emulsions with varied levels of fat, protein and water. Hardness, dry matter content and water drainage were determined by classical means and analysed for correlation with the NMR data with multivariate techniques. Partial least squares can calibrate and predict these properties directly from the continuous NMR exponential decays and yields regression coefficients higher than 82%. However, the calibration coefficients themselves belong to the continuous exponential domain and do little to explain the connection between NMR data and emulsion properties. Transformation of the NMR decays into a discreet domain with non-negative least squares permits the use of multilinear regression (MLR) on the resulting amplitudes as predictors and hardness or water drainage as responses. The MLR coefficients show that hardness is highly correlated with the components that have T2 distributions of about 20 and 200 ms whereas water drainage is correlated with components that have T2 distributions around 400 and 1800 ms. These T2 distributions very likely correlate with water populations present in pores with different sizes and/or wall mobility. The results for the emulsions studied demonstrate that NMR time domain decays can be employed to predict properties and to provide insight in the underlying microstructural features. PMID- 15833643 TI - Diffusion of polyethyleneglycols in casein solutions and gels as studied by pulsed field gradient NMR. AB - Molecular transport characterized by diffusion coefficients is a key feature of food processes and especially in dairy processes. Caseins represent 80% of the protein content in milk and are directly involved in the formation of dairy gels. Consequently, providing a quantitative description of the solute diffusion in casein gels should contribute significantly to rationalization of the dairy processes. The objectives of this study were to study the self-diffusion coefficient of molecular probes [polyethyleneglycol (PEGs)] in casein solutions and gels, in particular with regard to the effects of the probe molecular size and casein concentration. We have shown that the PEG diffusion was an efficient tool to reveal the structural information relative to the gel matrix structure. PMID- 15833644 TI - Displacement propagators of brine flowing within different types of sedimentary rock. AB - This paper explores the correlation between different microstructural characteristics of porous sedimentary rocks and the flow properties of a Newtonian infiltrating fluid. Preliminary results of displacement propagator measurements of brine solution flowing through two types of sedimentary rock cores are reported. The two types of rocks, Bentheimer and Portland, are characterized by different porosities, pore-size distributions and permeabilities. Propagators have been measured for brine flow rates of 1 and 5 ml/min. Significant differences are seen between the propagators recorded for the two rocks, and these are related to the spatial distribution of porosity within these porous media. PMID- 15833645 TI - Determination of water compartments in rat myocardium using combined D-T1 and T1 T2 experiments. AB - We have used combined D-T1 and T1-T2 correlation experiments to explore water compartments in rat heart tissue (myocardium). The results show that two main compartments can be identified, which we assign to extracellular (ec) and intracellular (ic) water. The exchange rate of water across the cell membrane was found to be on the order of 0.1 Hz. In addition, the T1-T2 correlation measurements indicate that the ic compartment contain two T2 populations. PMID- 15833646 TI - A comparison of experimental and simulated propagators in porous media using confocal laser scanning microscopy, lattice Boltzmann hydrodynamic simulations and nuclear magnetic resonance. AB - Confocal laser scanning microscopy has been used to obtain 3D optical image stacks of packings of glass ballotini in various fluorescent dye-containing fluids inside a 3D micromodel. The fluids' refractive index was matched to that of the glass ballotini so that clear images at an appreciable depth (approximately 400 microm) inside the packings were obtained. The lattice Boltzmann method was then used to produce 3D velocity fields through the 3D image stacks of the packed ballotini. These have been used in conjunction with a stochastic random-walk algorithm to produce simulated displacement propagators, which have been shown to be in qualitative agreement with experimental propagators, obtained using nuclear magnetic resonance, of water flowing through the exact same micromodel. PMID- 15833647 TI - Magnetic resonance imaging and relaxation analysis to predict noninvasively and nondestructively salt-to-moisture ratios in dry-cured meat. AB - The current systems are unable to control and predict the cured meat composition nondestructively and in a reasonable time for production needs. In this work, T1 and T2 maps were obtained, with a monoexponential model, on internal sections of Longissimus dorsi muscle at increasing salting times. The maps allow one to visualize the salting process nondestructively and noninvasively. The method goes beyond the simple qualitative visualization, because, for each section of the sample and in any region of the section, it is possible to obtain quantitative information on the progress of salting and to predict salt-to-moisture ratios. In addition, detailed relaxation measurements were performed on samples cored after imaging in order to define better the relaxation properties of the dry-cured meat. PMID- 15833648 TI - NMR-propagator measurements in porous media in the presence of surface relaxation and internal fields. AB - We measure the probability distribution P(zeta)--the propagator--of molecular displacements on Stokes flow through a pack of microporous glass beads and a carbonate rock. An optimized sampling of q-space is introduced for the measurement of a P(zeta) and its first moment zeta. Our results delineate and provide an understanding of the experimental regimes where background fields and surface relaxation distort the measured propagators. PMID- 15833649 TI - Quadrupolar interaction study of various cations confined in porous charged polymer film of sPI ionomers. AB - The structure of a sulfonated polyimide (sPI) ionomer membranes was investigated via the transport properties of various confined cations (7Li+, 23Na+, 87Rb+, 133Cs+). Their NMR spectra show large residual quadrupolar splitting depending on the orientation of the film in the static magnetic field B0. This behavior is the fingerprint of a macroscopic nematic ordering of charged interfaces. This is also confirmed by the anisotropy of the self-diffusion tensor measured by 1H and 7Li PGSE experiments on N(CH3)4+ and Li+ cations, respectively. PMID- 15833650 TI - MRI investigation and complementary numerical simulations of flow-through random bead packings with low aspect ratio. PMID- 15833651 TI - Micropore size analysis in oil-well cement by proton nuclear relaxation. AB - The durability of cement depends mainly on cement microstructure parameters such as pore size distribution. In this study, we have observed pore size distribution in a Class G oil-well cement immersed for 1 year in brine at T = 293 K, P = 10(5) Pa and in a down-hole condition at T = 353 K, P = 7.10(6) Pa by proton nuclear magnetic spin-lattice relaxation (1H-NMR). After 4 months of immersion at T = 293 K, P = 10(5) Pa, the distribution of discrete relaxation rates reveals the fractal feature of the pore size distribution, whereas after 1 year at T = 353 K, P = 7.10(6) Pa, a dispersed distribution is still observed. PMID- 15833652 TI - Structure characterization of foams and filled polymers by means of MRI. AB - The structure of foams and filled polymers can be analyzed by means of MRI. It is also possible to observe the deformation behavior of the structure of foams and filled polymers in situ. A displacement experiment (rigid body) was performed in the magnet and observed by MRI. The NMR images are analyzed by image processing. Average distances between particles are estimated by the use of the autocorrelation function and the spectrum of the autocorrelation function. The spectrum shows the spatial frequency of the distances. The displacement field was calculated by the cross-correlation function. Information about particle distances and micromechanical deformation can be obtained by NMR imaging methods by combining autocorrelation and cross-correlation. PMID- 15833653 TI - Field-gradient NMR diffusometry in poly(ethylene oxide) melts confined to nanoscopic pores of solid methacrylate matrices. AB - Depending on the choice of matrix constituents, the diameters of strands of linear, monodisperse poly(ethylene oxide) confined to nanoscopic pores of cross linked methacrylate matrices can be varied considerably. The samples were characterized by DSC, TEM, SEM and fringe field-gradient NMR diffusometry with respect to the strand diameter. A formalism evaluating diffusive spin echo attenuation curves based on the tube/reptation model allows the determination of the strand diameter. Values in the range 8-58 nm were found in accordance with TEM and SEM micrographs of shadow-cast freeze-fractured surfaces of the samples. PMID- 15833654 TI - Spatial distribution of coke residues in porous catalyst pellets analyzed by field-cycling relaxometry and parameter imaging. AB - The distribution of coke residues inside porous catalyst pellets was investigated on the molecular as well as the macroscopic scale. The presence of coke on the pore surface affects the relaxation properties of adsorbed liquid species; these were determined by field-cycling relaxometry for different polar and nonpolar liquids in metal-doped and metal-free catalyst carrier materials. The presence of metal in the Al2O3 matrix had only a minor influence on the dispersion behavior, while the interaction of the adsorbates with the coke layer leads to considerable changes in the relaxation times at low Larmor frequencies. Lowering the temperature to well below the bulk freezing point of dimethyl sulfoxide resulted in a slightly stronger frequency dependence of T1. Not only relaxation times but also the diffusion coefficient is affected by the presence of coke residues in the pores. For macroscopically heterogeneous samples, they offer the possibility to generate maps of the local coke concentration by introducing appropriate filters into NMR imaging sequences. High-temperature regeneration of coked catalysts leads to such heterogeneous distributions which is visualized by T1 parameter imaging. PMID- 15833655 TI - Ultrafast velocity imaging of single- and two-phase flows in a ceramic monolith. AB - Ceramic monoliths, comprising arrays of parallel channels, are increasingly being considered as an alternative to conventional packed beds for chemical processing operations involving both single- and two-phase flows. This paper reports results obtained using a technique based on the rapid acquisition with relaxation enhancement (RARE) pulse sequence in which multiple images are obtained from a single r.f. excitation. The technique is applied to study single- and two-phase flow in a monolith rated at 200 channels per square inch (cpsi). A single image frame, acquired in 156 ms, provides a characterization of the heterogeneity in the magnitude and direction of the flow within the monolith. PMID- 15833656 TI - Rapid imaging of fluid flow patterns in a narrow packed bed using MRI. AB - A gradient echo rapid velocity and acceleration imaging sequence (GERVAIS) has been developed and implemented to image liquid flow within a narrow packed bed. Two-dimensional velocity images have been acquired with an in-plane pixel size of 781 microm x 781 microm, with a data acquisition time of 20 ms for a single velocity component. Images of the x, y and z velocity vectors are reported. Data are reported for Reynolds numbers (based on particle diameter) of 200 and 300. In each case, GERVAIS images are compared with the results of a standard spin-echo phase-encoding velocity measurement. At Re = 200, steady-state flow is expected and the velocity images acquired using both techniques are consistent. At Re = 300, the GERVAIS sequence is able to image the unsteady-state flow field within this system. In contrast, the standard phase-encoding velocity measurement contains significant artefacts. PMID- 15833657 TI - Determination of porosity and flow distribution in packed beds by magnetic resonance imaging. AB - Magnetic resonance methods have been used to investigate the porosity and flow distributions in packed beds. The motivation of our study is to understand the behaviour of multiphase flow in fixed bed reactors, especially in trickle bed reactors. The results of this study should be included in a model for predicting the influence of hydrodynamics on hydrodesulfurization process in bench-scale reactors. PMID- 15833658 TI - Flow measurements below 50 mum: NMR microscopy experiments in lithographic model pore spaces. AB - Quasi two-dimensional random site percolation model objects have been prepared using a synchrotron radiation lithography technique with a spatial resolution better than 50 microm and an aspect ratio of up to 17. Flow of water through the pore space was studied with the aid of an NMR velocity mapping method and compared with a computational fluid dynamics simulation. In order to be able to measure and map widely distributed flow velocities with microscopic resolution (typically 40 x 40 microm), an experimental protocol that permits one to cover an effectively very wide velocity field of view (0.6-10 mm/s) had to be developed. PMID- 15833659 TI - Metal ion promoted hydrogels for bovine serum albumin adsorption: Cu(II) and Co(II) chelated poly[(N-vinylimidazole)-maleic acid]. AB - Poly[(N-vinylimidazole)-maleic acid] (poly(VIm-MA)), copolymeric hydrogels were prepared by gamma-irradiating ternary mixtures of N-vinylimidazole-maleic acid water in a (60)Co-gamma source. Cu(II) and Co(II) ions were chelated within the gels at pH=5.0. The maximum adsorption capacity of the gels were 3.71 mmol/g dry gel for Cu(II) and 1.25 mmol/g dry gel for Co(II) at pH=5.0. The swelling ratios of the gels were 1200% for poly(VIm-MA), 60 and 45% for Cu(II) and Co(II) chelated poly(VIm-MA) gels at pH=5.0 in acetate buffer solution. These affinity gels with different swelling ratios for plain poly(VIm-MA), Cu(II)-, and Co(II) chelated poly(VIm-MA), in acetate and phosphate buffers were used in the bovine serum albumin (BSA) adsorption/desorption studies in batch reactor. The maximum BSA adsorption capacities of the gels were 0.38 g/g dry gel for plain, 0.88 g/g dry gel for Cu(II)-chelated poly(VIm-MA) and 1.05 g/g dry gel for Co(II)-chelated poly(VIm-MA) gels. Adsorption capacity of BSA by the gels was reduced dramatically by increasing the ionic strength adjusted with NaCl. More than 95% of BSA were desorbed in 10 h in desorption medium containing 0.1M of EDTA for metal ion-chelated gels at pH=4.7. PMID- 15833660 TI - Interactions between hen egg-white lysozyme, PEG2,000, and PLA50 at the air-water interface. AB - In this paper, we compared the efficiency of polymer films, made of a poly(ethylene glycol) (PEG2,000)/poly(d,l-lactide) (PLA50) mixture, or a PEG2,000 PLA50 copolymer, to prevent adsorption of a model protein, the hen egg-white lysozyme (HEWL), at the air-water interface. This was achieved by analyzing the surface pressure/surface area curves, and the X-ray reflectivity data of the polymer films spread on a Langmuir trough, obtained in absence or in presence of the protein. For both the mixture and the copolymer, the amount of protein adsorbed at the air-water interface decreases when the density of the polymer surface coverage increases. It was shown that even in a condensed state, the polymer film made by the mixture can not totally prevent HEWL molecules to adsorb and penetrate the polymer mixed film, but however, protein molecules would not be directly exposed to the more hydrophobic phase, i.e. the air phase. It was also shown that the configuration adopted by the copolymer at the interface in its condensed state would prevent adsorption of HEWL molecules for several hours; this would be due in particular to the presence of PEG segments in the interfacial film. PMID- 15833661 TI - Changes of anti-oxidative enzymes and membrane peroxidation for soil water deficits among 10 wheat genotypes at seedling stage. AB - Drought is one of the major factors limiting crop production globally, with increasing global climate change making the situation more serious. Wheat is the staple food for more than 35% of world population, so wheat anti-drought physiology study is of importance to wheat production and biological breeding for the sake of coping with abiotic and biotic conditions. Much research is involved in this hot topic, but the pace of progress is not so large because of drought resistance being a multiple-gene-control quantitative character and wheat genome being larger (16,000 Mb). On the other hand, stress adaptive mechanisms are quite different, with stress degree, time course, materials, and experimental plots, thus increasing the complexity of the issue in question. Additionally, a little study is related to the whole life circle of wheat, which cannot provide a comprehensive understanding of its anti-drought machinery. We selected 10 kinds of wheat genotypes as materials, which have potential to be applied in practice, and measured relative change of anti-oxidative enzymes and membrane peroxidation through wheat whole growth-developmental circle (i.e. seedling, tillering and maturing). Here, we firstly reported the results of seedling stage as follows: (1) 10 wheat genotypes can be grouped into three kinds (A-C, respectively) according to their changing trend of the measured indices; (2) A performed better resistance drought under the condition of treatment level 1 (appropriate level), whose activities of anti-oxidative enzymes (POD, SOD, CAT) were higher and MDA lower and chlorophyll a+b higher; (3) B exhibited stronger anti-drought under treatment level 2 (light stress level), whose activities of anti-oxidative enzymes were higher, MDA lower and chlorophyll higher; (4) C expressed anti drought to some extent under treatment level 3 (serious stress), whose activities of anti-oxidative enzymes were stronger, MDA lower and chlorophyll higher; (5) these results demonstrated that different wheat genotypes have different physiological mechanisms to adapt themselves to changing drought stress, whose molecular basis is discrete gene expression profiling (transcriptom); (6) our results also showed that the concept accepted by most researchers, 70-75% QF is a proper supply for plants, was doubted, because this level could not reflect the true suitable level of wheat. The study in this respect is the key to wheat anti drought and biological saving-water; (7) our research can provide insights into physiological mechanisms of crop anti-drought and direct practical materials for wheat anti-drought breeding. PMID- 15833662 TI - Electroformation of giant liposomes from spin-coated films of lipids. AB - This paper describes spin-coating of solutions of lipids and using the resulting thin films for electroformation of giant liposomes. Spin-coating made it possible to generate uniform films of lipids with controllable thickness over large surfaces (>25 cm(2)) of indium tin oxide. Establishing a range of thicknesses optimal for electroformation (25-50 nm), we demonstrate formation of giant liposomes from lipids (such as asolectin, phosphatidylserine, and phosphatidylglycerol) that do not readily form giant liposomes from traditional, droplet-derived films. We compared liposomes from a spin-coated film of lipids to liposomes formed from traditional droplet-derived films and found that spin coated films produced larger (by factor of 2-5) and more abundant liposomes than droplet-derived films of lipids. Electroformation from spin-coated, homogenous lipid films of optimal thickness provided a reproducible way to obtain liposomes with diameters that are predominantly larger than 30 microm over the entire surface of formation. PMID- 15833663 TI - Giant vesicles formed by gentle hydration and electroformation: a comparison by fluorescence microscopy. AB - Giant unilamellar vesicles (diameter of a few tens of micrometers) are commonly produced by hydration of a dried lipidic film. After addition of the aqueous solution, two major protocols are used: (i) the gentle hydration method where the vesicles spontaneously form and (ii) the electroformation method where an ac electric field is applied. Electroformation is known to improve the rate of unilamellarity of the vesicles though it imposes more restricting conditions for the lipidic composition of the vesicles. Here we further characterize these methods by using fluorescence microscopy. It enables not only a sensitive detection of the defects but also an evaluation of the quantity of lipids in these defects. A classification of the defects is proposed and statistics of their relative importance in regard to both methods and lipid composition are presented: it shows for example that 80% of the vesicles obtained by electroformation from 98% 1,2-dioleoyl-sn-glycero-3-phosphocholine are devoid of significant defects against only 40% of the vesicles with the gentle hydration method. It is also shown that the presence of too many negatively charged lipids does not favor the formation of unilamellar vesicles with both methods. For the gentle hydration, we checked if the negatively charged lipids were inserted in the vesicles membrane in the same proportion as that of the lipid mixture from which they are formed. The constant incorporation of a negatively charged labeled lipid despite an increasing presence of negatively charged 1,2-dioleoyl-sn glycero-3-[phospho-l-serine] tends to confirm that the composition of vesicles is indeed close to that of the initial mixture. PMID- 15833664 TI - Extracting and analyzing sub-signals in heart rate variability. AB - A new statistical signal processing method, which was called independent component analysis (ICA) was used to extract sub-signals of heart rate variability (HRV). Ten healthy volunteers (4F, 6M) were involved in this study. Electrocardiogram (ECG) recording was consisted of 6min when the volunteer was lying and another 6min when the volunteer was standing. HRV was extracted from ECG. According to time-delay, HRV was divided into five groups as mixed signals. Five signals were reconstructed into two groups by ICA and the rebuilt two signals were transformed by Fourier transformation. Results showed that one group signal component centralized in low frequency (called IC1); the other did in high frequency (called IC2). The power of IC1 was significantly increased (P<0.05) while that of IC2 had no significant change (P>0.05) and the ratio of IC1 to total power was significantly increased (P<0.01) from lying to standing. Comparing the two postural results, it shows that IC1 may express sympathetic activity, and IC2 represents parasympathetic activity. Sympathetic and parasympathetic nervous function can be evaluated respectively and quantificationally by data and graphs from the two decomposed components. As an electro-physiological method, it can assist the investigation about the tension of autonomic nervous, myocardial bioelectricity activity, as well as myocardial cell membrane characters. PMID- 15833665 TI - Adsorption behaviors of poly(N-p-vinylbenzyl-4-o-beta-d-galactopyranosyl-[1-->4] d-gluconamide) by quartz-crystal microbalance. AB - Adsorption behaviors of amphiphilic poly(N-p-vinylbenzyl-4-o-beta-d galactopyranosyl-[1-->4]-d-gluconamide) (PVLA) on the polystyrene (PS) surface was studied using 27 MHz quartz-crystal microbalance (QCM). The amount of adsorbed PVLA on PS surface was increased with an increase of PVLA concentration as a Langmuir-type in a monolayer. The saturated mass change (DeltaM(max)) and association constant (K(a)) of PVLA on PS surface were 498.6 ng/cm(2) and 1.93 x 10(7)M(-1), respectively. The adsorbed PVLA on PS surface was specifically recognized by Allo A lectin due to specific interaction between galactose moieties in the PVLA and Allo A. The hydrophobic interaction between hydrophobic main chain of PVLA and hydrophobic surface of PS was reduced in the presence of urea and the diameter of PVLA aqueous solution was decreased with an increase of urea concentration. PMID- 15833666 TI - Entrapment of some compounds into biocompatible nano-sized particles and their releasing properties. AB - Two types of biocompatible nanoparticles with an average diameter of around 200 nm were formed only by mixing hydrolysates of chitosan and carboxymethyl cellulose (CMC). Nanoparticle A was produced from chitosanase hydrolysate of chitosan and cellulase hydrolysate of carboxymethyl cellulose, and nanoparticle B was produced from lysozyme hydrolysate of chitosan and the carboxymethyl cellulose hydrolysate. Negatively charged or amphoteric compounds were first mixed with chitosan hydrolysate and then added to carboxymethyl cellulose hydrolysate to effectively entrap them in the particles. Positively charged compounds could also be effectively entrapped by mixing the hydrolysates and the compound in the reverse order. Negatively charged compounds with high molecular weights were maintained in the particles even at the higher pH levels than the pK(a) of the amino groups of chitosan. Entrapped compounds were gradually released from nanoparticle A by lysozyme treatment. In contrast, there was no release from nanoparticle B. These results indicate that nanoparticle A can be applied to controlled-release drug delivery systems, and that nanoparticle B is stably retained in the body without releasing the entrapped compounds. PMID- 15833667 TI - Chitosan based surfactant polymers designed to improve blood compatibility on biomaterials. AB - We developed chitosan based surfactant polymers that could be used to modify the surface of existing biomaterials in order to improve their blood compatibility. These polymers consist of a chitosan backbone, PEG side chains to repel non specific protein adsorption, and hexanal side chains to facilitate adsorption and proper orientation onto a hydrophobic substrate via hydrophobic interactions. Since chitosan is a polycationic polymer, and it is thrombogenic, the surface charge was altered to determine the role of this charge in the hemocompatibility of chitosan. Charge had a notable effect on platelet adhesion. The platelet adhesion was greatest on the positively charged surface, and decreased by almost 50% with the neutralization of this charge. A chitosan surface containing the negatively charged SO(3)(-) exhibited the fewest number of adherent platelets of all surfaces tested. Coagulation activation was not altered by the neutralization of the positive charge, but a marked increase of approximately 5-6 min in the plasma recalcification time (PRT) was displayed with the addition of the negatively charged species. Polyethylene (PE) surfaces were modified with the chitosan surfactant resulting in a significant improvement in blood compatibility, which correlated to the increasing PEG content within the polymer. Adsorption of the chitosan surfactants onto PE resulted in approximately an 85 96% decrease in the number of adherent platelets. The surfactant polymers also reduced surface induced coagulation activation, which was indicated by the PEG density dependent increase in PRTs. These results indicate that surface modification with our chitosan based surfactant polymers successfully improves blood compatibility. Moreover, the inclusion of either negatively charged SO(3)( ) groups or a high density of large water-soluble PEG side chains produces a surface that may be suitable for cardiovascular applications. PMID- 15833668 TI - Cerebroside Langmuir monolayers originated from the echinoderms I. Binary systems of cerebrosides and phospholipids. AB - The surface pressure (pi)-area (A), the surface potential (DeltaV)-A and the dipole moment (mu( perpendicular))-A isotherms were obtained for two-component monolayers of two different cerebrosides (LMC-1 and LMC-2) with phospholipids of dipalmitoylphosphatidylcholine (DPPC) and with dipalmitoylphosphatidylethanolamine (DPPE) on a subphase of 0.5 M sodium chloride solution as a function of phospholipid compositions by employing the Langmuir method, the ionizing electrode method, and the fluorescence microscopy. Surface potentials (DeltaV) of pure components were analyzed using the three-layer model proposed by Demchak and Fort. The contributions of the hydrophilic saccharide group and the head group to the vertical component of the dipole moment (mu( perpendicular)) were estimated. The miscibility of cerebroside and phospholipid in the two-component monolayers was examined by plotting the variation of the molecular area and the surface potential as a function of the phospholipid molar fraction (X(phospholipid)), using the additivity rule. From the A-X(phospholipid) and DeltaV(m)-X(phospholipid) plots, partial molecular surface area (PMA) and apparent partial molecular surface potential (APSP) were determined at the discrete surface pressure. The PMA and APSP with the mole fraction were extensively discussed for the miscible system. Judging from the two-dimensional phase diagrams, these can be classified into two types. The first is a positive azeotropic type; the combinations of cerebrosides with DPPC are miscible with each other. The second is a completely immiscible type: the combination of cerebrosides with DPPE. Furthermore, a regular surface mixture, for which the Joos equation was used for the analysis of the collapse pressure of two-component monolayers, allowed calculation of the interaction parameter (xi) and the interaction energy (-Delta epsilon) between the cerebrosides and DPPC component. The miscibility of cerebroside and phospholipid components in the monolayer state was also supported by fluorescence microscopy. PMID- 15833669 TI - Cerebroside Langmuir monolayers originated from the echinoderms: II. Binary systems of cerebrosides and steroids. AB - Two-component Langmuir monolayers formed on a subphase of 0.5M sodium chloride solution were investigated for two different cerebrosides (LMC-1 and LMC-2) with steroids of cholesterol (Ch) and cholesteryl sodium sulfate (Ch-S); i.e. LMC 1/Ch, LMC-1/Ch-S, LMC-2/Ch, and LMC-2/Ch-S were examined in terms of surface pressure (pi), the surface potential (DeltaV) and the dipole moment (mu( perpendicular)) as a function of surface area (A) by employing the Langmuir method, the ionizing electrode method, and the fluorescence microscopy. Surface potentials (DeltaV) of steroids were analyzed using the three-layer model proposed by Demchak and Fort. The miscibility of cerebrosides and steroids in the insoluble monolayers was examined by plotting the variation of the molecular area and the surface potential as a function of the steroid molar fraction (X(steroid)) based upon the additivity rule. From the A-X(steroid) and DeltaV(m) X(steroid) plots, partial molecular surface area (PMA) and apparent partial molecular surface potential (APSP) were determined at the different surface pressures. The PMA and APSP with the mole fraction were discussed for the miscible system. Judging from the two-dimensional phase diagrams, they can be classified into two types. The first is a completely immiscible type; the combination of cerebrosides with cholesterol. The second is a negative azeotropic type, where cerebrosides and cholesteryl sodium sulfate are completely miscible both in the expanded state and in the condensed state. In addition, a regular surface mixture (the Joos equation for the analysis of the collapse pressure of two-component monolayers) allowed calculation of the interaction parameter (xi) and the interaction energy (-Delta epsilon) between the cerebrosides and Ch-S. The miscibility of cerebroside and steroid components in the monolayer state was also supported by fluorescence microscopy. PMID- 15833670 TI - Endocrine and metabolic disturbances in critically ill patients: To intervene or not? PMID- 15833671 TI - Case reports-something everyone wants read, but nobody wants to read. How to get them published! PMID- 15833672 TI - Endocrine interventions in the ICU. AB - Patients with critical illness, particularly those who depend on intensive care for a prolonged period of time, have a high morbidity and mortality. The acute and chronic phases of critical illness are associated with distinct endocrine alterations. Acute endocrine adaptations to the severe stress of critical illness, comprising an activated anterior pituitary function, have been selected by nature and can, as such, be considered as beneficial for surviving. These adaptations disappear or wane during the prolonged phase of critical illness. In this phase there is a reduced pulsatile secretion of different anterior pituitary hormones and the so-called "wasting syndrome" occurs. This prolonged endocrine/metabolic stress response is quite different from the acute response and may, to some extent, no longer be adaptive. Intervention within the endocrine system, however, remains highly controversial, as it is difficult to differentiate between beneficial adaptations and harmful abnormalities and to outline strategies for therapy. Insulin infusion titrated to maintain normoglycemia may be a notable exception, as this intervention has been proven to increase survival and reduce morbidity of surgical intensive care patients. Treatment of "relative adrenal failure" with hydrocortisone also appears to improve the outcome of patients with septic shock, but diagnostic and dosing issues still remain unresolved. Although extensive research has shown that infusion of hypothalamic-releasing peptides is able to restore physiological hormonal patterns within the somatotropic, thyrotropic, and gonadal axes and, thereby, to generate a controlled anabolic response, further research is needed to investigate whether such interventions actually improve the outcome of critical illness. PMID- 15833673 TI - Growth factors and their relationship to neoplastic and paraneoplastic disease. AB - Growth factors are extracellular signaling molecules that act in an autocrine and paracrine fashion to regulate growth, proliferation, differentiation, and survival of cells. Dysregulation of the growth factor networks is intimately related to the molecular pathogenesis of neoplastic and paraneoplastic disease. Increasing knowledge of the molecular mechanisms underlying growth factors and their actions on cell cycling, cell division, and cell death is shedding light on new therapeutic avenues for molecular targeting of tumors. Epidermal growth factor and vascular endothelial growth factor both offer examples of how growth factor biology and its relationship to cancer can be harnessed to create effective clinical therapeutic tools such as monoclonal antibodies. This approach heralds a future in which rational molecular oncological therapy may increasingly become the norm. PMID- 15833674 TI - The physician as comforter. AB - Patients, their families and even physicians themselves have commented on the lack of communication skills, empathy and concern for the psychosocial aspect of patient care among doctors of today. The call by many in the medical community for a renewed commitment to professionalism is partially in response to these concerns. Recently, medical ethics has focused on technical legal issues but ethics also has much to contribute on issues relating to the doctor-patient relationship. By analyzing traditional religious attitudes toward this relationship one can develop universal insights that still have much relevance to modern medicine for physicians and patients of all faiths. PMID- 15833675 TI - How many patients in atrial fibrillation admitted to an acute medical unit will benefit from oral anticoagulation? Application of the results of the major randomized controlled trials to 141 consecutive, unselected, elderly patients using a decision support computer program. AB - BACKGROUND: Although several randomized, control trials (RTC) suggest that oral anticoagulation (OAC) benefits patients with atrial fibrillation (AF), this might not be true for hospitalized patients with co-morbid conditions. If the results of the RTCs are valid, then how many patients in AF admitted to an acute medical unit will benefit from OAC? METHODS: An RCT-based decision analysis model calculated the quality-adjusted life expectancy (QALE) gain from OAC for 141 unselected consecutive patients over 65 years of age with AF admitted to an acute medical unit. RESULTS: If treated with aspirin, all 141 patients were predicted to gain QALE compared with placebo. If the quality of life adjustment (QoLA) on OAC was the same as placebo, then 104 patients were predicted to benefit from OAC compared with aspirin, while 63 patients were predicted to benefit at a QoLA of 0.99 (overall benefit 0.13+/-0.15 QALYs, range 0.01-0.88 QALYs). These 63 patients were more likely to have had a stroke, diabetes, hypertension, heart failure or heart attack, and less likely to have impaired renal function than those predicted not to benefit. The 78 patients predicted not to benefit from OAC included 11 younger patients without heart failure, hypertension, diabetes or cerebrovascular disease; the remaining 67 patients, however, were older, more likely to have heart failure and/or renal impairment and were at high risk of both stroke and bleeding. CONCLUSION: An RCT-based decision analysis model suggests that more than half the patients in AF admitted to a small rural hospital with acute medical conditions are unlikely to benefit from OAC, while all will benefit from aspirin. PMID- 15833676 TI - Evaluation of health-related quality-of-life and dyspnea scales in patients with idiopathic pulmonary fibrosis. Correlation with pulmonary function tests. AB - BACKGROUND: Idiopathic pulmonary fibrosis (IPF) has a profound impact on the functional status of patients. We sought to determine whether the health-related quality of life is affected in patients with IPF. METHODS: A total of 25 patients with IPF (aged 41-72 years) and 30 healthy subjects were evaluated in a cross sectional study. All subjects were asked to complete three dyspnea scales-the Borg (BORG), the oxygen-cost diagram (OCD) and the modified MRC questionnaire-as well as a number of health-related quality-of-life (HRQoL) questionnaires, i.e., the Saint-George Respiratory Questionnaire (SGRQ), the Quality of Well-Being (QWB) and the Hospital Anxiety and Depression Questionnaire (HAD). Pulmonary function tests, arterial blood gas measurements at rest and during exercise, chest radiographs and the duration of disease were used for correlation. RESULTS: The mean total values of all of the scales used were near the lower best values of each scale in IPF patients and were significantly different from values in the control group. Lung volumes (FVC and TLC) correlated significantly with the SGRQ. All dyspnea scales (BORG, OCD and MRC) showed a significant correlation with diffusing capacity and blood oxygenation at rest and during exercise. The duration of the disease correlated with all dyspnea scales used and with the SGRQ and HAD Questionnaires. CONCLUSIONS: Our results suggest that dyspnea scales and the SGRQ are sensitive tools for assessing health-related quality of life in patients with IPF. PMID- 15833677 TI - Increased incidence of hepatocellular carcinoma (HCC) in HIV-1 infected patients. AB - BACKGROUND: The likely increased incidence of hepatocarcinoma (HCC) in HIV-1 infected patients has not yet been demonstrated. METHODS: We studied all cases of HCC occurring in HIV-1 infected patients in our hospital during the past 15 years. Incidence and survival time were compared with those of the general population in the same area and the same time of the study. RESULTS: We found 6 cases of HCC in a cohort of 2383 HIV-1 infected patients between 1986 and 2001. This is a higher than expected incidence rate of HCC compared with the general population, with a standardized incidence ratio of 13.95. Chronic hepatitis virus infection and alcohol abuse were present in four and two cases, respectively. In one patient, no liver disease was known before the HCC and the surrounding liver was normal in the necropsy study. CONCLUSION: The improved survival of patients on highly active antiretroviral treatment (HAART) and the increasing incidence of end-stage liver disease in these patients caused by chronic hepatitis virus infection and alcohol abuse may be responsible for an increase in the incidence of HCC in HIV-1 infected patients. PMID- 15833678 TI - Influence of old age on survival after prolonged mechanical ventilation. AB - BACKGROUND: While the proportion of elderly people in society is increasing, little is known about the influence of old age on survival after prolonged mechanical ventilation. METHODS: A retrospective follow-up study of 120 patients who had been mechanically ventilated for at least 10 days was performed with a follow-up of up to 5 years. In-hospital survival and post-hospital survival were documented. Also, the functional status of survivors was recorded. The predictive value of age and several other clinical and laboratory parameters for outcome was analyzed. RESULTS: The in-hospital survival of the 120 patients studied was 35%. While age below 50 years was associated with improved survival, age was not predictive of in-hospital survival for patients over 50 years of age. The post hospital survival was 77% 1 year after discharge, with almost all survivors being functionally independent. Again, old age was not predictive of survival. CONCLUSIONS: For patients aged 50 years or older, patient age does not predict survival after prolonged mechanical ventilation. Even very old survivors have a reasonable life expectancy and regain full functional status. PMID- 15833679 TI - Severe transient neutropenia associated with acute human immunodeficiency virus type 1 infection. AB - Severe neutropenia associated with primary HIV infection is unusual. We report the fifth case in a 50-year-old male with a neutrophil count of 500/mm(3) and a platelet count of 92,000/mm(3) at the time of early HIV-1 seroconversion. In all previously published cases and in our case, severe neutropenia was a very early sign of acute HIV infection, and it regressed spontaneously and quickly. HIV testing should be recommended when severe neutropenia is observed, especially in the context of a flu-like or mononucleosis-like infectious syndrome. PMID- 15833680 TI - Coronary stent bacterial infection with multiple organ septic emboli. AB - We describe an 80-year-old patient who developed Staphylococcus aureus septicemia several days after the implantation of a double stent in the proximal and mid left anterior descending artery. The infection was complicated by multiple abscesses in the lungs and liver, as well as by bilateral bacterial endophthalmitis requiring right vitrectomy. Long-term antibiotic treatment was successful. Rarity notwithstanding, heightened awareness of this potential complication of a common cardiac procedure is important since diagnosis and immediate therapy are mandatory. PMID- 15833681 TI - Is there any medical therapeutic option in hepatopulmonary syndrome? A case report. AB - Hepatopulmonary syndrome (HPS) is a well-known complication characterized by abnormalities of arterial oxygenation in patients with chronic liver disease, with or without portal hypertension. There is no definitive treatment for HPS except liver transplantation. We report on a patient with HBV-related liver cirrhosis and HPS who received garlic and lamivudine for 3 years. Signs of liver failure and hypoxemia gradually improved, indicating that lamivudine may improve the functional reserve of the liver, while garlic may help to reduce the signs and symptoms of HPS. PMID- 15833682 TI - Pleural effusion and thickening due to cabergoline use in a patient with Parkinson's disease. AB - A 62-year-old Caucasian man with a history of Parkinson's disease developed bilateral pleural thickening and a large, right-sided, exudative pleural effusion. Further investigations revealed a chronic inflammatory process involving the pleura as a result of exposure to cabergoline used to treat the Parkinson's disease. The inflammatory process reversed after the discontinuation of cabergoline. PMID- 15833683 TI - Horton's disease (temporal arteritis) associated with antineutrophil cytoplasmic antibodies (ANCA) in a demented elderly subject. PMID- 15833684 TI - Acute motor sensory axonal Guillain-Barre syndrome and myasthenia gravis. PMID- 15833685 TI - Metastatic right atrial mass. PMID- 15833687 TI - Eosinophilic gastroenteritis. AB - Eosinophilic gastroenteritis despite its uncommon occurrence is one of the most important primary eosinophilic gastrointestinal disorders, and most commonly presents with abdominal pain. The terminology is, however, misleading because all levels of the gastrointestinal tract from the esophagus to the rectum may be affected. A history of atopy and allergies is present in 25-75% cases. The heterogeneity in the clinical presentations of EG is determined by the site and depth of eosinophilic infiltration. Eosinophilic intestinal inflammation also occurs secondarily in the gastrointestinal tract in inflammatory bowel disease, autoimmune diseases, as reactions to medications, infections, hypereosinophilia syndrome, and after solid organ transplantation. Recent investigations providing an insight into the pathogenesis of eosinophilic gastroenteritis support a critical role for allergens, eosinophils, Th-2 type cytokines, and eotaxin in mediating eosinophilic inflammation. The diagnosis is confirmed by demonstrating prominent tissue eosinophilia on histopathology. Treatment recommendations based on data extrapolated from retrospective, uncontrolled studies, and expert opinion support the use of restricted diets, corticosteroids, leukotriene receptor antagonists, and mast cell stabilizers. Many unanswered questions remain with regard to the natural history, optimal duration of therapy, safer steroid-sparing long-term treatment agents, and the means of reliable and non-invasive follow-up. PMID- 15833688 TI - Familial Mediterranean fever--a not so unusual cause of abdominal pain. AB - Familial Mediterranean fever is a hereditary syndrome characterised by recurrent episodes of fever and serositis, resulting in pain in the abdomen, chest, joints and muscles. It is primarily diagnosed in people of Jewish, Arabic, Turkish or Armenian ancestry and is caused by mutations in the gene encoding for pyrin. Abdominal FMF attacks resemble the clinical presentation of 'acute abdomen', with severe abdominal pain and rigidity, but in FMF symptoms always resolve spontaneously. It is important to distinguish these regular pain episodes from small bowel obstruction due to adhesions to prevent life-threatening bowel strangulation. In most cases, colchicine will prevent new painful attacks. This seminar also discusses other causes of abdominal pain in FMF patients. PMID- 15833689 TI - Vasculitis in the gastrointestinal tract. AB - The primary systemic vasculitides are a rare group of inflammatory disorders, where damage is directed against the blood vessels. They range in severity from acute self-limiting illness to chronic life and organ threatening diseases requiring long-term treatment with toxic immunosuppressive regimes. None of them have specific tests and few have hard diagnostic criteria. This review summarises the gastrointestinal features of these diseases, along with their general clinical and epidemiological features and the evidence available to guide in their management. PMID- 15833690 TI - Acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is characterised by neurovisceral crises the most common clinical presentation of which is abdominal pain. It is an autosomal dominant condition with incomplete penetrance and is potentially life threatening. The key point in management is to suspect and confirm the diagnosis as early as possible in order to treat the attack and to avoid inappropriate treatments which may exacerbate the crisis. In this chapter we briefly outline the haem biosynthetic pathway and how deficiencies in individual enzymes give rise to the different porphyrias. We then describe the clinical features and diagnosis of AIP, followed by a discussion of pathogenesis, highlighting advances in the molecular biology of AIP and introducing the debate as to whether neurovisceral crises might result from porphyrin precursor neurotoxicity or from haem deficiency. Finally we discuss management, including family screening, avoidance of triggering factors, analgesia, maintenance of a high calorie intake, and administration of haem derivatives. PMID- 15833691 TI - Deficiencies of C1 inhibitor. AB - Hereditary and acquired deficiencies of the C1 inhibitor result in a single prominent symptom, namely angioedema. Angioedema may involve the skin, the gastrointestinal tract or the upper airway. Genetically determined defects in C1INH cause hereditary angioedema. The defect may be acquired as the result of an auto-antibody to C1INH or be due to the generation of anti-idiotypic antibody to monoclonal immunoglobulins as occurs in various B cell lymphoproliferative diseases. Androgens provide prophylaxis against attacks of angioedema. There is no widely approved treatment for acute attacks of angioedema although several promising drugs are now in the final stages of clinical trials. PMID- 15833692 TI - Abdominal epilepsy. AB - Abdominal epilepsy is an uncommon syndrome in which gastrointestinal complaints, most commonly abdominal pain, result from seizure activity. It is characterized by (1) otherwise unexplained, paroxysmal gastrointestinal complaints, (2) symptoms of a central nervous system disturbance, (3) an abnormal electroencephalogram with findings specific for a seizure disorder, and (4) improvement with anticonvulsant medication. We review the history of the syndrome and analyze all 36 cases reported in the English literature from the last 34 years. The most common gastrointestinal symptoms include abdominal pain, nausea and vomiting, while the most common neurological symptoms include lethargy and confusion. After exclusion of more common etiologies for the presenting complaints, workup should proceed with an electroencephalogram. Where the diagnosis is seriously considered, neurological consultation should be considered. Treatment typically begins with anticonvulsant medication, and resolution of symptoms with therapy helps to confirm the diagnosis. PMID- 15833693 TI - Diabetic thoracic polyradiculopathy. AB - Diabetic thoracic polyradiculopathy usually causes severe, chronic abdominal pain in patients with type 2 diabetes of variable duration. Other diabetic complications, weight loss and paretic abdominal wall protrusion are common. Sensory, motor and autonomic functions are affected. The diagnosis can be made from the characteristic history, physical examination findings, paraspinal electromyography, and other procedures. The differential diagnosis includes postherpetic neuralgia, abdominal wall pain, malignancy, and other spinal disorders. The pathology appears to be immune-mediated neurovasculitis resulting in ischemic injury. Traditional therapy is symptomatic, but recent pathological findings and clinical experience suggest that immunotherapy may be effective. PMID- 15833694 TI - Chronic mesenteric ischemia. AB - Chronic mesenteric ischemia is an unusual but important cause of abdominal pain. Although this condition accounts for only 5% of all intestinal ischemic events, it can have significant clinical consequences. Among its many causes, atherosclerotic occlusion or severe stenosis is the most common. This disorder has an indolent course that results in extensive collateral vascular formation. Thus, symptoms occur when at least two of the three main splanchnic vessels are affected. Intestinal angina, weight loss, and sitophobia are common clinical features. Diagnosis can often be made by noninvasive methods such as computerised axial tomographic angiography, magnetic resonance angiography, and duplex ultrasonography as well as by invasive catheter angiography. Therapy of chronic mesenteric ischemia depends on the extent and location of vascular disease. Alternatives to traditional surgical bypass are becoming more common including embolectomy, thrombolysis, and percutaneous angioplasty with vascular stenting. Early intervention is vital as the natural course of this illness can be debilitating. Furthermore, this has potential to develop into life-threatening acute mesenteric ischemia with subsequent bowel infarction and death. Long-term studies have shown that the risk of developing symptoms from asymptomatic but significant mesenteric vascular disease is 86% with overall 40% mortality rate. The recognition and management of this unusual but important cause of abdominal pain is discussed in detail in this review. PMID- 15833695 TI - Unusual causes of abdominal pain: sickle cell anemia. AB - Sickle cell disease is characterized by chronic hemolytic anemia and vaso occlusive painful crises. The vascular occlusion in sickle cell disease is a complex process and accounts for the majority of the clinical manifestation of the disease. Abdominal pain is an important component of vaso-occlusive painful crises. It often represents a substantial diagnostic challenge in this population of patients. These episodes are often attributed to micro-vessel occlusion and infarcts of mesentery and abdominal viscera. Abdominal pain due to sickle cell vaso-occlusive crisis is often indistinguishable from an acute intra-abdominal disease process such as acute cholecystitis, acute pancreatitis, hepatic infarction, ischemic colitis and acute appendicitis. In the majority of cases, however, no specific cause is identified and spontaneous resolution occurs. This chapter will focus on etiologies, pathophysiology and management of abdominal pain in patients with sickle cell disease. PMID- 15833696 TI - Proceedings of the International Workshop on Surface Modification for Chemical and Biochemical Sensing. November 13-16, 2003. Bialowieza, Poland. PMID- 15833697 TI - Modified electrodes based on lipidic cubic phases. AB - The lipidic cubic phase can be characterized as a curved bilayer forming a three dimensional, crystallographical, well-ordered structure that is interwoven by aqueous channels. It provides a stable, well-organized environment in which diffusion of both water-soluble and lipid-soluble compounds can take place. Cubic phases based on monoacylglycerols form readily and attract our interest due to their ability to incorporate and stabilize proteins. Their lyotropic and thermotropic phase behaviour has been thoroughly investigated. At hydration over 20%, lipidic cubic phases Ia3d and Pn3m are formed. The latter is stable in the presence of excess water, which is important when the cubic phase is considered as an electrode-modifying material. Due to high viscosity, the cubic phases can be simply smeared over solid substrates such as electrodes and used to host enzymes and synthetic catalysts, leading to new types of catalytically active modified electrodes as shown for the determination of cholesterol, CO(2), or oxygen. The efficiency of transport of small hydrophilic molecules within the film can be determined by voltametry using two types of electrodes: a normal-size electrode working in the linear diffusion regime, and an ultramicroelectrode working under spherical diffusion conditions. This allows determining both the concentration and diffusion coefficient of the electrochemically active probe in the cubic phase. The monoolein-based cubic phase matrices are useful for immobilizing enzymes on the electrode surface (e.g., laccases from Trametes sp. and Rhus vernicifera were employed for monitoring dioxygen). The electronic contact between the electrode and the enzyme was maintained using suitable electroactive probes. PMID- 15833698 TI - Design, synthesis and characterization of monomolecular interfacial layers. AB - We have designed a series of monomolecular films comprised of four basic structural motifs. We have used these films for a variety of purposes, ranging from support structures for chromophore arrays to the creation of selective and biomimetic interfaces. We will discuss the several different types of interfacial binding chemistry that are used in the construction of these interfacial films, for both monomeric and polymeric layer structures. Following a discussion of the construction of the adlayers, we describe several uses of these assemblies, in areas ranging from adsorption to optical signal processing, and the formation of biomimetic interfacial structures. PMID- 15833699 TI - Electrochemistry of cytochrome c incorporated in Langmuir-Blodgett films of Nafion and Eastman AQ 55. AB - Ultrathin films of Nafion and Eastman-AQ 55 loaded with cytochrome c (cyt c) were obtained and transferred on indium tin oxide (ITO) electrodes via the Langmuir Blodgett (LB) technique. The pressure-area isotherms for mixed ionomer-protein films indicate that the miscibility of cyt c in the interfacial layer is better for Nafion than for AQ 55. Interestingly, these composite films maintain the electroactivity of cyt c without requiring the addition of promoters or mediators. Both for AQ 55-cyt c and Nafion-cyt c films, the half-wave potential for the reversible reduction of ferricytochrome c corresponds to the value expected for the weakly adsorbed protein. The modified electrodes show electrocatalytic reaction with ascorbate anion. Comparison with previous literature reports indicate that for Nafion the LB coating procedure is unique in keeping the electroactivity of cyt c. PMID- 15833700 TI - Immobilization and electrochemical redox behavior of cytochrome c on fullerene film-modified electrodes. AB - Two different fullerene film-modified electrodes were prepared and used for surface immobilization and electrochemical property investigation of horse heart cytochrome c (cyt c). Both a pristine fullerene film and fullerene-palladium (C(60)-Pd) polymer film-modified platinum, glassy carbon and indium-tin-oxide (ITO) electrodes were used. The immobilized cyt c was characterized by piezoelectric microgravimetry at a quartz crystal microbalance (QCM), UV-visible absorption, and X-ray photoelectron spectroscopy (XPS), as well as cyclic voltammetry (CV) techniques. The UV-visible spectral studies revealed a small blue shift of both the Soret and Q band of the heme moiety of cyt c, immobilized on the C(60)-Pd polymer film-modified ITO electrode, as compared to the bands of cyt c in solution suggesting that molecules of cyt c are densely packed onto the surface of the modified electrode. The CV studies revealed a quasi-reversible electrode behavior of the heme moiety indicating the occurrence of kinetically hindered electron transfer. A good agreement was found between the values of cyt c electrode surface coverage determined by piezoelectric microgravimetry and cyclic voltammetry. For piezoelectric microgravimetry, these values ranged from 0.5 x 10(-10) to 2.5 x 10(-10) mol cm(-2), depending upon the amount of cyt c present in solution and the time allowed for immobilization, which compared with a value of 3.6+/-0.4 x 10(-10) mol cm(-2) determined by CV. The possible mechanisms of cyt c immobilization on the C(60) film and C(60)-Pd film-modified electrodes are also discussed. PMID- 15833701 TI - TiO2 phytate films as hosts and conduits for cytochrome c electrochemistry. AB - Cytochrome c is accumulated into a film of TiO(2) nanoparticles and phytate by adsorption from an aqueous solution into the mesoporous structure. Stable voltammetric responses and high concentrations of redox protein within the TiO(2) phytate layer can be achieved. Two types of electrode systems are reported with (i) the modified TiO(2) phytate film between electrode and aqueous solution phase and (ii) the modified TiO(2) phytate film buried under a porous gold electrode ('porotrode'). The electrical conductivity of TiO(2) phytate films is measured and compared in the dry and in the wet state. Although in the dry state essentially insulating, the TiO(2) phytate film turns into an electrical conductor (with approximately 4 Omega cm specific resistivity assuming ohmic behaviour) when immersed in aqueous 0.1 M phosphate buffer solution at pH 7. The redox protein cytochrome c is therefore directly connected to the electrode via diffusion and migration of electrons in the three dimensional mesoporous TiO(2) phytate host structure. Electron transfer from cytochrome c to TiO(2) is proposed to be the rate-determining step for this conduction mechanism. PMID- 15833702 TI - Integrated circuits protection with the Langmuir-Blodgett Films. AB - Integrated circuits (ICs) can be protected from the environment with the encapsulating polymer layer. Protection properties of such a polymer barrier layer depend strongly from the structure of thin region where polymer stays in direct contact with the IC surface. One of the interesting questions is how thick should this interphase film be to assure good environmental protection to the IC conductor lines, preventing from their corrosion and failure. In order to answer this question a set of electronic testers with Al conductor lines were modified with 1, 20, 50 multilayers of stearic acid molecules deposited in the Langmuir Blodgett (LB) transfer method. Next, the electronic testers were subjected to the highly accelerated aging conditions (100% relative humidity (RH), 100 degrees C) for a period of up to 800 h and conductor lines resistivity changes were monitored. Electronic testers modified with 20 multilayers of stearic acid were better protected from the accelerated aging conditions than the testers modified with 1 monolayer or 50 multilayers. Obtained results suggest that the thickness of the interphase region separating IC surface and polymeric film should be in the range of 10 nm. PMID- 15833703 TI - Direct electrochemistry of heme multicofactor-containing enzymes on alkanethiol modified gold electrodes. AB - Direct electrochemistry of heme multicofactor-containing enzymes, e.g., microbial theophylline oxidase (ThOx) and D-fructose dehydrogenase (FDH) from Gluconobacter industrius was studied on alkanethiol-modified gold electrodes and was compared with that of some previously studied complex heme enzymes, specifically, cellobiose dehydrogenase (CDH) and sulphite oxidase (SOx). The formal redox potentials for enzymes in direct electronic communication varied for ThOx from 112 to -101 mV (vs. Ag|AgCl), at pH 7.0, and for FDH from -158 to -89 mV, at pH 5.0 and pH 4.0, respectively, on differently charged alkanethiol layers. Direct and mediated by cytochrome c electrochemistry of FDH correlated with the existence of two active centres in the protein structure, i.e., the heme and the pyrroloquinoline quinone (PQQ) prosthetic groups. The effect of the alkanethiols of different polarity and charge on the surface properties of the gold electrodes necessary for adsorption and orientation of ThOx, FDH, CDH and SOx, favourable for the efficient electrode-enzyme electron transfer reaction, is discussed. PMID- 15833704 TI - The effect of support material and conditioning on wettability of PAH/PSS multilayer films. AB - An efficient method for characterizing wetting properties of heterogeneous surfaces produced by sequential adsorption of polyelectrolytes was developed. Three types of polyelectrolytes were used: polyallylamine hydrochloride (PAH), polyethyleneimine (PEI), both of a cationic type, and polysodium 4 styrenesulfonate (PSS), of an anionic type. Multilayer films were prepared by 'layer-by-layer' (LbL) deposition technique. Natural ruby mica, glass, titanium foil and silicon wafers were used as the support material for PE films. Wetting of polyelectrolyte films was determined experimentally by contact angle measurements, using technique of direct image analysis of shape of sessile drops. Periodic oscillations in contact angle values were observed for multilayers terminated by polycation and polyanion, respectively, and the variations in contact angle values strongly depended on the conditions of adsorption and multilayer treatment after deposition. Therefore, the influence of ionic strength of polyelectrolyte solution used for deposition on wetting of multilayer films was considered and also the effect of conditioning in different environments was investigated. It is usually assumed that film properties and stability strongly depend on the first layer which is used to anchor a multilayer at the surface of support material. To investigate influence of the first layer, PAH/PSS films were compared with more complex ones having PEI as the first layer with a sequence of PSS/PAH deposited on top of it. PMID- 15833705 TI - Spectroscopic and electrochemical characterization of interfacial biomimetic assemblies on electrochemically generated gold oxide surfaces. AB - We report on the formation of a gold oxide layer and the reaction of this oxide with an acid chloride to form a stable, relatively complete monolayer bound through an ester-like bond to the gold oxide surface. We have used cyclic voltammetry, FTIR and optical ellipsometry to characterize this novel monolayer structure. The exposed functional groups of this monolayer can participate in subsequent surface reactions, opening the door to the use of oxide-based surface attachment chemistry on metallic substrates. This chemistry will allow for the formation of films tailored to contain both hydrophobic and hydrophilic regions, stacked at predetermined distances from the substrate that may serve as biomembrane mimetic assemblies. PMID- 15833706 TI - Polyoxometallates as inorganic templates for electrocatalytic network films of ultra-thin conducting polymers and platinum nanoparticles. AB - We develop a concept of fabrication of the multilayer network films on electrodes by exploring the ability of a Keggin-type polyoxometallate, phosphododecamolybdate (PMo(12)O(40)(3-)), to form stable anionic monolayers (templates) on carbon and metals including platinum. By repeated alternate treatments in the solution of PMo(12)O(40)(3-) (or in the colloidal suspension of polyoxometallate-protected Pt-nanoparticles) and in the solution of monomer (e.g. anilinium) cations, the amount of the material can be increased systematically (layer-by-layer) to form stable three-dimensional assemblies on electrode (e.g. glassy carbon) surfaces. In the resulting hybrid (organic-inorganic) films, the layers of negatively charged polyoxometallate, or polyoxometallate-protected (stabilized) Pt-nanoparticles, are linked or electrostatically attracted by ultra thin layers of such positively charged conducting polymers as polyaniline (PANI), polypyrrole (PPy) or poly(3,4-ethylenedioxythiophene), PEDOT. Consequently, the attractive physicochemical properties of polymers and reactivity of polyoxometallate or noble metal particles are combined. The films are functionalized and show electrocatalytic properties towards reduction of nitrite, bromate, hydrogen peroxide or oxygen. They are of importance to the chemical and biochemical sensing as well as to the biochemical and medical applications. PMID- 15833707 TI - Surface immobilized optical probes: pyrene molecules covalently attached to silica and indium-doped tin oxide. AB - We report on the covalent attachment of pyrene to quartz and indium-doped tin oxide (ITO) surfaces, and its characterization by steady-state and time-resolved emission spectroscopy. We estimate the chromophore surface coverage on the basis of their electrochemical oxidation mechanism in acidic solution. Electrochemical oxidation of immobilized pyrene results in the formation of radical cations, which are converted into hydroxypyrene and, after a second oxidation step, are transformed into isomeric forms of dihydroxypyrene/pyrenedione redox couples. Our findings demonstrate that surface-bound pyrene can be used as a spectral probe in environments that are isolated from oxidizing conditions and agents. PMID- 15833708 TI - Ion equilibrium in lichen surrounding. AB - In laboratory conditions, the ionic equilibriums between a solution and a cation active layer of epiphytic lichens Hypogymnia physodes immersed in the solution were examined. It was found that such equilibriums, due to exchange of mobile cations: H, Na, K, Ca, and Mg, are established in natural conditions between a lichen and atmospheric water. PMID- 15833709 TI - Electrochemical nitric oxide sensor preparation: a comparison of two electrochemical methods of electrode surface modification. AB - Platinum electrodes modified with Mn(II) 5-(N-(8-pyrrole-yl-3,6-dioxa-1 aminooctane)phenylamide-10,15,20-trimethoxyphenylporphyrin (Mn(II)triOMeTCPPyP) using multi-sweep cyclic voltammetry and differential pulse amperometry were evaluated as electrocatalytic surfaces for the oxidation of nitric oxide. The electrodes modified using the pulse amperometric approach were more sensitive towards the detection of nitric oxide. The increased sensitivity led to the attainment of a wider linear dynamic range for the quantification of nitric oxide. PMID- 15833710 TI - Antibody immobilisation on the metal and silicon surfaces. The use of self assembled layers and specific receptors. AB - The use of Staphylococcal protein A and lectins as intermediate immobilising agents allows operators to orient antibodies (Ab) towards the solution due to the presence of a specific binding sites of immunoglobulin (Ig) molecules. Antibodies of different species of animals have unequal affinities to individual lectins. The effective thickness of immobilised Ab's depends on the type of substrates used and increases in the following sequence: bare gold or silicon surface, the surface treated with self-assembled polyelectrolytes (PESA) or with protein A or some lectins deposited on the preliminary formed polyelectrolyte layer. The glycolysated protein of jp51 may be selectively immobilised from the mixture of retroviral proteins (p24 and jp51), if it is necessary to distinguish infected animals from preliminarily immunised ones by means of a vaccine based on p24 protein. It was shown that the use of Staphylococcal protein A, instead of some lectins as intermediate layer for the Ab immobilisation, does not lead to a more sensitive determination of such low-weight toxins as 2,4-dichlorophenoxyacetic acid (2,4-D). The above-mentioned results were obtained with surface plasmon resonance (SPR) technique. PMID- 15833711 TI - DNA imaged on a HOPG electrode surface by AFM with controlled potential. AB - Single-molecule AFM imaging of single-stranded and double-stranded DNA molecules self-assembled from solution onto a HOPG electrode surface is reported. The interaction of DNA with the hydrophobic surface induced DNA aggregation, overlapping, intra- and intermolecular interactions. Controlling the electrode potential and using the phase images as a control method, to confirm the correct topographical characterization, offers the possibility to enlarge the capability of AFM imaging of DNA immobilized onto conducting substrates, such as HOPG. The application of a potential of +300 mV (versus AgQRE) to the HOPG enhanced the robustness and stability of the adsorbed DNA molecules, increasing the electrostatic interaction between the positively charged electrode surface and the negatively charged DNA sugar-phosphate backbone. PMID- 15833712 TI - Kinetic study of the degradation of a potential local anesthetic drug in serum using the DNA-based electrochemical biosensor. AB - DNA-drug association interaction at the DNA modified screen-printed electrode for 1-methyl-2-piperidinoethylester of 2-hexoxyphenylcarbamic acid was found leading to an accumulation of the drug within the DNA layer. A procedure for the determination of drug in blood serum matrix using the protein precipitation and voltammetric measurement of the electroactive drug with the DNA biosensor was obtained and an effort was done to apply it for an assay of the drug enzymatic degradation in human and rabbit sera at 37 degrees C. PMID- 15833713 TI - Direct immobilisation of DNA probes for the development of affinity biosensors. AB - An immobilisation procedure based on the direct coupling of thiolated probes (Probe-C6-SH) to bare gold sensor surfaces has been compared with a reference immobilisation method, based on the coupling of biotinylated probes onto a streptavidin-coated dextran-modified surface. The instrumentations used were a quartz crystal microbalance (QCM) and the optical instruments Biacore X and Spreeta based on surface plasmon resonance (SPR). The performances of the DNA based sensors resulting from direct coupling of thiolated DNA probes onto electrodes of quartz crystals or gold SPR-chips have been studied in terms of the main analytical parameters, i.e. selectivity, sensitivity, reproducibility, etc. In particular, the two immobilisation approaches have been applied to the analysis of oligonucleotides, DNA amplified by polymerase chain reaction (PCR) and genomic DNA enzymatically digested. PMID- 15833714 TI - Detection of DNA hybridization by ABEI electrochemiluminescence in DNA-chip compatible assembly. AB - The electrochemiluminescence (ECL) of a luminol derivate (ABEI) generated both by a carbon electrode and a polypyrrole-coated carbon electrode was examined. It was found that the polypyrrole film (ppy) did not inhibit the ECL. After that, ABEI anchored on a single stranded DNA target (ODNt) has been used for the ECL detection of the hybridization between a complementary single stranded DNA probe (ODNp) covalently linked to a polypyrrole support and the ODNt. The ECL detection has been performed using a DNA sensor having a low surface concentration of ODNp probes, constituted of a polypyrrole copolymer electrosynthesized from a pyrrole ODNp/pyrrole monomer ratio of 1/20,000. PMID- 15833715 TI - C/EBPdelta and C/EBPgamma bind the CCAAT-box in the human beta-globin promoter and modulate the activity of the CACC-box binding protein, EKLF. AB - Developmental- and tissue-specific expression of globin genes is mediated by a few key elements within the proximal promoter of each gene. DNA-binding assays previously identified NF-Y, GATA-1, C/EBPbeta and C/EBPgamma as candidate regulators of beta-globin transcription via the CCAAT-box, a promoter element situated between CACC- and TATA-boxes. We have identified C/EBPdelta as an additional beta-globin CCAAT-box binding protein. In reporter assays, we show that C/EBPdelta can co-operate with EKLF, a CACC-box binding protein, to activate the beta-globin promoter, whereas C/EBPgamma inhibits the transcriptional activity of EKLF in this assay. PMID- 15833716 TI - Bacillus subtilis tRNA(Pro) with the anticodon mo5UGG can recognize the codon CCC. AB - In Bacillus subtilis, four codons, CCU, CCC, CCA, and CCG, are used for proline. There exists, however, only one proline-specific tRNA having the anticodon mo(5)UGG. Here, we found that this tRNA(Pro)(mo(5)UGG) can read not only the codons CCA, CCG and CCU but also CCC, using an in vitro assay system. This means that the first nucleoside of its anticodon, 5-methoxyuridine (mo(5)U), recognizes A, G, U and C. On the other hand, it was reported that mo(5)U at the first position of the anticodon of tRNA(Val)(mo(5)UAC) can recognize A, G, and U but not C. A comparison of the structure of the anticodon stem and loop of tRNA(Pro)(mo(5)UGG) with those of other tRNAs containing mo(5)U at the first positions of the anticodons suggests that a modification of nucleoside 32 to pseudouridine (Psi) enables tRNA(Pro)(mo(5)UGG) to read the CCC codon. PMID- 15833717 TI - The effects of group dynamics on resident progress committee deliberations. AB - BACKGROUND: Prior investigations suggest that resident progress decisions by committee provide a broader perspective on performance and result in less grade inflation. There is countervailing concern that group dynamics may compromise quality of progress decisions. PURPOSE: To determine whether and how group dynamics compromise decision making about resident progress. METHODS: Researchers recorded and analyzed participant comments during a resident progress committee meeting. End-of-rotation (EOR) evaluations were analyzed and compared to progress committee meeting results. RESULTS: EOR and progress committee comments were similar in content. The ratio of specific to general comments was higher for EOR evaluations (5:1) than for progress committee meetings (2:1). EOR evaluation comments provided more supporting evidence for assertions. Individual progress committee participants did not dominate discussion or sway decision making. Participant progress committee meeting comments were consistent with their EOR comments failing to support the presence of progress committee meeting "feeding frenzies." CONCLUSIONS: Results suggest that progress committee meeting group dynamics do not seriously compromise the validity of resident progress decisions. PMID- 15833718 TI - Psychometric characteristics of a write-up assessment form in a medicine core clerkship. AB - BACKGROUND: Case write ups are ubiquitous in medicine clerkships, yet few studies have examined how they are assessed. PURPOSE: To examine the feasibility, reliability, and validity of scores on a new write-up assessment form. METHODS: In 2002, medicine core clerkship students (n = 165) submitted 3 patient write ups (n = 493). Each was graded using a 14-item form and given a Global Write-Up Assessment. Final Global Write-Up grades were correlated to National Board of Medical Examiners medicine subject exam scores and multiple clerkship ratings. RESULTS: For most items on the form, the full 4-point rating range was used. The reproducibility coefficient was .67 (SE = .12). Final Global Write-Up grades were correlated with exam scores (r = .35, p < .05) and inpatient (r = .28, p < .05) and outpatient (r = .16, p < .05) course grades. CONCLUSIONS: The write-up assessment form, as used in a design with multiple write ups and raters, provides relatively precise estimates of performance and can be used to assess written documentation skills. PMID- 15833719 TI - Does a high tech (computerized, animated, PowerPoint) presentation increase retention of material compared to a low tech (black on clear overheads) presentation? AB - BACKGROUND: Computerized digital presentations have become standard in most medical schools without research proving their effectiveness. PURPOSE: The purpose was to determine if differences in (a) subjective evaluation; (b) short term retention of material; and (c) long-term retention of material occurred with the use of static overheads versus computerized, animated PowerPoint for a presentation to medical students. METHODS: All students rotating on a required clerkship attended a standard lecture presented by 1 faculty member. The content of the presentation remained the same, but the instructional media varied (overheads on even months, animated PowerPoint on odd months). Students completed a posttest and subjective evaluation immediately following the lecture and repeated the posttest 1 year later. Unpaired t tests were used to compare the 2 groups. RESULTS: There were no significant differences between the groups on any parameter. CONCLUSIONS: In this study, students rated both types of presentation equally and displayed no differences in short- or long-term retention of material. PMID- 15833720 TI - A comparison of in-class learner engagement across lecture, problem-based learning, and team learning using the STROBE classroom observation tool. AB - BACKGROUND: Having recently introduced team learning into the preclinical medical curriculum, evidence of the relative impact of this instructional method on in class learner engagement was sought. PURPOSE: To compare patterns of engagement behaviors among learners in class sessions across 3 distinct instructional methods: lecture, problem-based learning (PBL), and team learning. METHODS: Trained observers used the STROBE classroom observation tool to measure learner engagement in 7 lecture, 4 PBL, and 3 team learning classrooms over a 12-month period. Proportions of different types of engagement behaviors were compared using chi-square. RESULTS: In PBL and team learning, the amount of learner-to learner engagement was similar and much greater than in lecture, where most engagement was of the learner-to-instructor and self-engagement types. Also, learner-to-instructor engagement appeared greater in team learning than in PBL. CONCLUSIONS: Observed engagement behaviors confirm the potential of team learning to foster engagement similar to PBL, but with greater faculty input. PMID- 15833721 TI - Measuring teaching effectiveness in a pre-clinical multi-instructor course: a case study in the development and application of a brief instructor rating scale. AB - BACKGROUND: Despite widespread use, misunderstandings persist about student evaluations of teaching. These evaluations have not been well examined in the common medical school setting of the multi-instructor, preclinical lecture course. PURPOSE: The study evaluated the psychometrics of a brief student evaluation of a teaching instrument developed for a multi-instructor 2nd-year course and described its application. METHODS: An 11-item instrument was developed and administered to 276 students to evaluate 27 lecturers per year in 3 years of an introductory clinical psychiatry course. A fully crossed research design allowed for a thorough analysis of variability in ratings. RESULTS: Generalizability analysis showed good reliability and relatively large Student x Lecturer interactions. Profile analysis generated distinct lecturer teaching profiles. CONCLUSIONS: Judicious use of a psychometrically sound student evaluation of a teaching instrument can be used to assist faculty and course development. Administering the evaluation instrument to an entire class produces no better reliability than administration to randomly selected subgroups of students. PMID- 15833722 TI - Teaching evidence-based medicine to internal medicine residents: the efficacy of conferences versus small-group discussion. AB - BACKGROUND: Evidence-based medicine (EBM) is a required component of the Accreditation Council for Graduate Medical Education's Practice-Based Learning core competency. PURPOSE: To compare the efficacy of conferences and small-group discussions in enhancing EBM competency. METHODS: EBM conferences and small-group discussions were integrated into an internal medicine curriculum. EBM competency was assessed by written examination following participation in both groups and compared with residents who had not participated in either format. RESULTS: Small group discussion participants (n = 10) scored higher on an EBM exam (17.8 +/- 4.5 correct out of 25) when compared with 10 conference participants (12.2 +/- 4.6, p = .010) and 26 residents who did not participate in either format (12.0 +/- 4.5, p = .002). Small-group discussion participants also reported increased confidence and high satisfaction. CONCLUSIONS: Although more resource intensive, small-group discussions resulted in increased EBM knowledge, increased confidence with critical appraisal skills, and high satisfaction compared with a conference-based format. PMID- 15833723 TI - Facilitation of problem finding among first year medical school students undergoing problem-based learning. AB - BACKGROUND: Adaptation to problem-based learning (PBL) is a difficult process for high school graduates who are not used to self-directed learning, especially in the freshmen year of medical school. The difficulty includes finding problems from a given case. PURPOSE: Evaluate the effect of an intervention to facilitate case-based problem finding among medical school freshmen undergoing a PBL tutorial. METHODS: Medical school freshmen in 2000 (nonintervened group) and 2001 (intervened group) participated in the study. The intervened group received the modified problem-based program by (a) having briefings on the importance of problem finding, (b) encouragement by the tutors in problem finding, and (c) reinforcement using a self-assessment sheet. At the end of the year, the ability of students to extract problems from a short case was evaluated and compared with the nonintervened students. RESULTS: The intervened group extracted a significantly greater number of problems than the nonintervened group. When extracted problems were categorized, the intervened group was able to generate more questions in a greater number of specified categories. CONCLUSIONS: Interventions to foster problem finding significantly facilitated acquisition of problem extraction skills among young medical students. PMID- 15833724 TI - Development of an advocacy curriculum in a pediatric residency program. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education now requires pediatric residency programs to include advocacy training, but few guidelines for development exist. PURPOSE: To determine faculty and resident attitudes regarding advocacy training, and to develop consensus on the best method to teach and learn advocacy. METHODS: Pediatric residents (n = 28) and faculty (n = 51) participated in a survey about experience and training in advocacy, identified issues important to address in an advocacy curriculum, and rated a variety of learning methods. RESULTS: Nearly 90% of residents felt advocacy training was needed, and 82% felt advocacy activities would continue following residency. Faculty and residents agreed that participation in short-term or longitudinal projects was the best way to teach and learn advocacy skills. Respondents identified and ranked 45 issues to address through advocacy efforts and training, and these results were used to develop and implement an advocacy curriculum. CONCLUSIONS: Residents and faculty were interested in incorporating an advocacy curriculum, and survey results were used to guide curriculum development. PMID- 15833725 TI - A comparison of statistical criteria for setting optimally discriminating MCAT and GPA thresholds in medical school admissions. AB - BACKGROUND: Using Medical College Admission Test-grade point average (MCAT-GPA) scores as a threshold has the potential to address issues raised in recent Supreme Court cases, but it introduces complicated methodological issues for medical school admissions. PURPOSE: To assess various statistical indexes to determine optimally discriminating thresholds for MCAT-GPA scores. METHODS: Entering classes from 1992 through 1998 (N = 752) are used to develop guidelines for cut scores that optimize discrimination between students who pass and do not pass the United States Medical Licensing Examination (USMLE) Step 1 on the first attempt. RESULTS: Risk differences, odds ratios, sensitivity, and specificity discriminated best for setting thresholds. Compensatory versus noncompensatory procedures both accounted for 54% of Step 1 failures, but demanded different performance requirements (noncompensatory MCAT-biological sciences = 8, physical sciences = 7, verbal reasoning = 7--sum of scores = 22; compensatory MCAT total = 24). CONCLUSIONS: Rational and defensible intellectual achievement thresholds that are likely to comply with recent Supreme Court decisions can be set from MCAT scores and GPAs. PMID- 15833726 TI - Impact of a residency research program on research activity, faculty involvement, and institutional cost. AB - BACKGROUND: The impact of residency research programs on resident research activity, faculty involvement, and institutional cost has not been well described. DESCRIPTION: A strategy to increase resident research activity was implemented in a community-based internal medicine residency program. Strategy components included a resident research director, a research elective, cost reimbursement, and a research requirement. Associated outcomes of research activity, faculty involvement, and institutional cost are described. EVALUATION: The annual number of research submissions increased from 0 to 39 over 6 years. The greatest increase in number of research submissions was seen following the dual implementation of the cost reimbursement and research requirement interventions. The annual number of faculty coauthors rose from 0 to 24 in 6 years. Average cost per accepted project was US 1,023.00 dollars. CONCLUSION: The strategy described was associated with a marked increase in resident research activity and faculty involvement. The cost of supporting resident research activity is significant. PMID- 15833727 TI - Utilization and value of personal digital assistants on an epidemiology final examination. AB - BACKGROUND: The utility of personal digital assistants (PDAs) in basic science medical education is uncertain. DESCRIPTION: Student outcomes on an epidemiology course final examination for academic years 2003 and 2004 were examined. Students were given permission to use PDAs on the final examination, and self-selected whether these instruments were used. Performance on the examination based on use of a PDA and whether students thought it was useful for the examination was compared. EVALUATION: A total of 389 students took the final examination, with an 88% response rate to the survey questions. No statistically significant differences were found on final examination scores. No trends toward significance were found on analyses of the total examination, specific topical domains, or on specific questions where a PDA might be expected to be especially useful. CONCLUSIONS: From this study, it can be concluded that use of PDAs and whether students thought they might be helpful had no measurable effect on performance on an epidemiology final examination. Further delineation of the possible use of PDAs in a basic science course and on the final examination is indicated. PMID- 15833728 TI - Teaching about family violence: a proposed model curriculum. AB - BACKGROUND: Medical schools around the country have initiated ad hoc teaching around family violence issues, especially child abuse and intimate partner violence (IPV). However, these activities typically are not integrated with other aspects of the curriculum, or with each other. Consequently, students do not have the opportunity to learn the broader issues related to family violence prevention and intervention. DESCRIPTION: To address these concerns, students at the Southern Illinois University (SIU) School of Medicine participated in an elective to develop a model family violence curriculum encompassing child abuse, IPV, sexual assault, and elder abuse. The activities use standardized patients and resource people, including hotline and shelter staff, legal professionals, and law enforcement. EVALUATION: This article describes the family violence teaching activities at SIU preceding Curriculum 2000 and the proposed model curriculum including principles guiding the curriculum, the curriculum sequence, and descriptions of the proposed patient cases and related activities. CONCLUSIONS: Family violence teaching in medical schools is merely a first step. Continuing medical education in violence-related issues must also be available. PMID- 15833729 TI - Medical students' perception of good PBL tutors in Taiwan. AB - BACKGROUND: Problem-based learning (PBL) approach for medical education has been in use in Western countries for more than 30 years, but its use in Asian countries is quite recent. Because of cultural differences, the perception of a good tutor from the perspectives of students may be different. DESCRIPTION: Students from a PBL curriculum medical school at Fu-Jen Catholic University, Taiwan, were asked to provide a narrative evaluation of tutor performance from tutorial learning. Twenty-nine tutors and their 49 undergraduate students participated in the study from September 2002 to June 2003. EVALUATION: Qualitative methods were used to identify and code the tutors' behaviors. Students preferred tutors who (a) had knowledge in both basic and clinical science areas, (b) had appropriate facilitative tutorial skills, and (c) had positive personality traits. CONCLUSION: The results showed that students valued personality aspects of a tutor, an area that had been neglected in previous studies. PMID- 15833730 TI - Pimping: perspectives of 4th year medical students. AB - BACKGROUND: A well-known phenomenon among U.S. medical students known as pimping, or the pedagogical device of questioning students in the clinical setting, receives virtually no attention in medical literature. PURPOSE: Identifying 4th year medical students' relevant knowledge and attitudes about pimping may help educators understand the range of beliefs about pimping and the role it plays in the socialization process into the medical community. METHODS: Over a 2-month period, 11 fourth-year medical students at a Midwest medical school were asked 6 open-ended questions focusing on pimping as understood and experienced in the clinical setting. Investigators individually analyzed the interview data using qualitative methods to characterize students' experiences and recurring ideas and concepts. RESULTS: All students noted the hierarchical nature of pimping, viewing it as a tool for attendings or residents to assess students' levels of knowledge. Although some students experienced malignant pimping, humiliated by incessant questioning or questions inappropriate to their level of training, all the students in the sample were positive about pimping and its effectiveness as a pedagogical tool. Investigators found that location within the clinical setting determines how students define and understand the motives for pimping. CONCLUSIONS: Understanding how students define and experience the pimping phenomenon prepares medical educators to scrutinize pimping as a pedagogical tool and to provide the most effective and encouraging environment for students. PMID- 15833731 TI - What does performance on a standardized test mean in relation to patient experiences in different venues? PMID- 15833733 TI - Carboxyl tail cysteine mutants of the thyrotropin-releasing hormone receptor type 1 exhibit constitutive signaling: role of palmitoylation. AB - We studied the role of carboxyl tail cysteine residues and their palmitoylation in constitutive signaling by the thyrotropin-releasing hormone (TRH) receptor type 1 (TRH-R1) in transfected mammalian cells and in Xenopus laevis oocytes. To study palmitoylation, we inserted a factor Xa cleavage site within the third extracellular loop of TRH-R1, added a carboxyl-terminal C9 immunotag and expressed the mutant receptor in Chinese hamster ovary cells. We identified TRH R1-specific palmitoylation in the transmembrane helix-7/carboxyl-tail receptor fragment mainly at Cys-335 and Cys-337. In contrast to a mutant truncated at Cys 335 that was reported previously to be constitutively active, a receptor truncated at Lys-338 (K338Stop), which preserves Cys-335 and Cys-337, and C337Stop and N336Stop, which preserve Cys-335, did not exhibit increased constitutive signaling. TRH-R1 mutants substituted singly by Gly or Ser at Cys 335 or Cys-337 did not exhibit constitutive signaling. By contrast, substitution of both cysteines (C335G/C337G or C335S/C337S) yielded TRH-R1 mutants that exhibited marked constitutive signaling in mammalian cells. In the oocyte, constitutive signaling by C335G/C337G resulted in homologous (of C335G/C337G) and heterologous (of M1 muscarinic receptor) desensitization. Because both Cys-335 and Cys-337 have to be substituted or deleted for constitutive signaling, we propose that a single palmitoylation site in the proximal carboxyl tail is sufficient to constrain TRH-R1 in an inactive conformation. PMID- 15833734 TI - The amino acid Asn136 in HIV-1 reverse transcriptase (RT) maintains efficient association of both RT subunits and enables the rational design of novel RT inhibitors. AB - The highly conserved Asn136 is in close proximity to the nonnucleoside reverse transcriptase (RT) inhibitor (NNRTI)-specific lipophilic pocket of human immunodeficiency virus type 1 (HIV-1) RT. Site-directed mutagenesis has revealed that the catalytic activity of HIV-1 RT mutated at position Asn136 is heavily compromised. Only 0.07 to 2.1% of wild-type activity is retained, depending on the nature of the amino acid change at position 136. The detrimental effect of the mutations at position 136 occurred when the mutated amino acid was present in the p51 subunit but not in the p66 subunit of the p51/p66 RT heterodimer. All mutant enzymes could be inhibited by second-generation NNRTIs such as efavirenz. They were also markedly more sensitive to the inactivating (denaturating) effect of urea than wild-type RT, and the degree of increased urea sensitivity was highly correlated with the degree of (lower) catalytic activity of the mutant enzymes. Replacing wild-type Asn136 in HIV-1 RT with other amino acids resulted in notably increased amounts of free p51 and p66 monomers. Our findings identify a structural/functional role for Asn136 in stabilization of the RT p66/p51 dimer and provide hints for the rational design of novel NNRTIs or drugs targeting either Asn136 in the beta7-beta8 loop of p51 or its anchoring point on p66 (the peptide backbone of His96) so as to interfere with the RT dimerization process and/or with the structural support that the p51 subunit provides to the p66 subunit and which is essential for the catalytic enzyme activity. PMID- 15833735 TI - Alpha 1 subunit-containing GABA type A receptors in forebrain contribute to the effect of inhaled anesthetics on conditioned fear. AB - Inhaled anesthetics are believed to produce anesthesia by their actions on ion channels. Because inhaled anesthetics robustly enhance GABA A receptor (GABA(A) R) responses to GABA, these receptors are considered prime targets of anesthetic action. However, the importance of GABA(A)-Rs and individual GABA(A)-R subunits to specific anesthetic-induced behavioral effects in the intact animal is unknown. We hypothesized that inhaled anesthetics produce amnesia, as assessed by loss of fear conditioning, by acting on the forebrain GABA(A)-Rs that harbor the alpha1 subunit. To test this, we used global knockout mice that completely lack the alpha1 subunit and forebrain-specific, conditional knockout mice that lack the alpha1 subunit only in the hippocampus, cortex, and amygdala. Both knockout mice were 75 to 145% less sensitive to the amnestic effects of the inhaled anesthetic isoflurane. These results indicate that alpha1-containing GABA(A)-Rs in the hippocampus, amygdala, and/or cortex influence the amnestic effects of inhaled anesthetics and may be an important molecular target of the drug isoflurane. PMID- 15833736 TI - Heme oxygenase-1 gene activation by the NAD(P)H oxidase inhibitor 4-(2 aminoethyl) benzenesulfonyl fluoride via a protein kinase B, p38-dependent signaling pathway in monocytes. AB - Heme oxygenase (HO)-1 is the inducible isoform of the rate-limiting enzyme of heme degradation and modulates the inflammatory immune response. Because HO-1 is up-regulated by NAD(P)H oxidase activators such as lipopolysaccharide and 12-O tetradecanoylphorbol-13-acetate in monocytic cells, we investigated the gene regulation of HO-1 by the chemical NAD(P)H oxidase inhibitor 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF). Unexpectedly, AEBSF induced endogenous gene expression and promoter activity of HO-1 in cell cultures of human and mouse monocytes. Inhibition of the phosphatidylinositol 3-kinase/protein kinase B (PKB) pathway by pharmacological inhibitors and cotransfection of an expression vector for a dominant negative mutant of PKB reduced the AEBSF-dependent induction of HO 1 gene transcription. Accordingly, overexpressed constitutively active PKB markedly up-regulated HO-1 promoter activity. AEBSF activated the mitogen activated protein kinases (MAPK) JNK and p38. Inhibition of p38alpha and p38beta, but not that of JNK or p38gamma and p38delta, prevented the induction of HO-1 gene expression by AEBSF. p38 was stimulated by AEBSF in a PKB-dependent manner as demonstrated by a luciferase assay with a Gal4-CHOP fusion protein. Finally, AEBSF- and PKB-dependent induction of HO-1 promoter activity was reduced by simultaneous mutation of an E-box motif (-47/-42) and a cAMP response element/AP 1 element (-664/-657) of the proximal HO-1 gene promoter. Overexpression of the basic helix-loop-helix transcription factor USF2 and coactivator p300 enhanced the AEBSF-dependent response of the HO-1 promoter. The data suggest that the transcriptional induction of HO-1 gene expression by AEBSF is mediated via activation of a PKB, p38 MAPK signaling pathway. PMID- 15833737 TI - Interleukin-1beta differentially regulates beta2 adrenoreceptor and prostaglandin E2-mediated cAMP accumulation and chloride efflux from Calu-3 bronchial epithelial cells. Role of receptor changes, adenylyl cyclase, cyclo-oxygenase 2, and protein kinase A. AB - Here we tested the effect of interleukin-1beta, a pro-inflammatory cytokine, on cAMP accumulation and chloride efflux in Calu-3 airway epithelial cells in response to ligands binding to adenylyl cyclase-coupled receptors such as the beta2 adrenoreceptor and EP prostanoid receptors. Interleukin-1beta significantly increased isoprenaline-induced cAMP accumulation by increasing beta2 adrenoreceptor numbers via a protein kinase A-dependent mechanism. In contrast, interleukin-1beta significantly impaired prostaglandin E2-induced cAMP accumulation by induction of cyclo-oxygenase-2, prostaglandin E2 production, and a resulting down-regulation of adenylyl cyclase. The cAMP changes were all mirrored by alterations in chloride efflux assessed using the fluorescent chloride probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide with interleukin-1beta increasing chloride efflux in response to isoprenaline and reducing the response to prostaglandin E2. Studies with glibenclamide confirmed that chloride efflux was via the cystic fibrosis transmembrane conductance regulator. Calu-3 expresses EP4 receptors, but not EP2, and receptor expression is reduced by interleukin-1beta. Collectively, these results provide mechanistic insight into how interleukin-1beta can differentially regulate cAMP generation and chloride efflux in response to different adenylyl cyclase-coupled ligands in the same cell. These findings have important implications for diseases involving inflammation and abnormal ion flux such as cystic fibrosis. PMID- 15833738 TI - A novel monothiol glutaredoxin (Grx4) from Escherichia coli can serve as a substrate for thioredoxin reductase. AB - Glutaredoxins are ubiquitous proteins that catalyze the reduction of disulfides via reduced glutathione (GSH). Escherichia coli has three glutaredoxins (Grx1, Grx2, and Grx3), all containing the classic dithiol active site CPYC. We report the cloning, expression, and characterization of a novel monothiol E. coli glutaredoxin, which we name glutaredoxin 4 (Grx4). The protein consists of 115 amino acids (12.7 kDa), has a monothiol (CGFS) potential active site and shows high sequence homology to the other monothiol glutaredoxins and especially to yeast Grx5. Experiments with gene knock-out techniques showed that the reading frame encoding Grx4 was essential. Grx4 was inactive as a GSH-disulfide oxidoreductase in a standard glutaredoxin assay with GSH and hydroxyethyl disulfide in a complete system with NADPH and glutathione reductase. An engineered CGFC active site mutant did not gain activity either. Grx4 in reduced form contained three thiols, and treatment with oxidized GSH resulted in glutathionylation and formation of a disulfide. Remarkably, this disulfide of Grx4 was a direct substrate for NADPH and E. coli thioredoxin reductase, whereas the mixed disulfide was reduced by Grx1. Reduced Grx4 showed the potential to transfer electrons to oxidized E. coli Grx1 and Grx3. Grx4 is highly abundant (750-2000 ng/mg of total soluble protein), as determined by a specific enzyme link immunosorbent assay, and most likely regulated by guanosine 3',5' tetraphosphate upon entry to stationary phase. Grx4 was highly elevated upon iron depletion, suggesting an iron-related function for the protein. PMID- 15833739 TI - Prostaglandin E2 stimulates fibronectin expression through EP1 receptor, phospholipase C, protein kinase Calpha, and c-Src pathway in primary cultured rat osteoblasts. AB - Fibronectin (Fn) is involved in the early stages of bone formation, and prostaglandin E (PGE) is an important factor regulating osteogenesis. Here we found that PGE(2) enhanced extracellular Fn assembly in rat primary osteoblasts, as shown by immunofluorescence staining and enzyme-linked immunosorbent assay. PGE(2) also increased the protein levels of Fn by using Western blotting analysis. By using pharmacological inhibitors or activators or genetic inhibition by the EP receptor, antisense oligonucleotides revealed that the EP(1) receptor but not other PGE receptors is involved in PGE(2)-mediated up-regulation of Fn. At the mechanistic level, Ca(2+) chelator (1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester)), phosphatidylinositol phospholipase C inhibitor (U73122), or Src inhibitor (PP2) attenuated the PGE(2) induced Fn expression. Protein kinase C (PKC) inhibitor (GF109203X) also inhibited the potentiating action of PGE(2). Furthermore, treatment with antisense oligonucleotides of various PKC isoforms, including alpha, beta, epsilon, and delta, demonstrated that alpha isozyme plays an important role in the enhancement action of PGE(2) on Fn assembly. Flow cytometry and reverse transcription-PCR showed that PGE(2) and 17-phenyl trinor PGE(2) (EP(1)/EP(3) agonist) increased the surface expression and mRNA level of alpha5 or beta1 integrins. Fn promoter activity was enhanced by PGE(2) and 17-phenyl trinor PGE(2) in cells transfected with pGL2F1900-Luc. Cotransfection with dominant negative mutants of PKCalpha or c-Src inhibited the potentiating action of PGE(2) on Fn promoter activity. Local administration of PGE(2) or 17-phenyl trinor PGE(2) into the metaphysis of the tibia via the implantation of a needle cannula significantly increased the Fn and alpha5beta1 integrin immunostaining and bone volume of secondary spongiosa in tibia. Taken together, our results provided evidence that PGE(2) increased Fn and promoted bone formation in rat osteoblasts via the EP(1)/phospholipase C/PKCalpha/c-Src signaling pathway. PMID- 15833740 TI - Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration. AB - The human immunodeficiency virus-1 (HIV-1) envelope glycoprotein (Env) is comprised of non-covalently associated gp120/gp41 subunits that form trimeric spikes on the virion surface. Upon binding to host cells, Env undergoes a series of structural transitions, leading to gp41 rearrangement necessary for fusion of viral and host membranes. Until now, the prefusion state of gp41 ectodomain (e gp41) has eluded molecular and structural analysis, and thus assessment of the potential of such an e-gp41 conformer to elicit neutralizing antibodies has not been possible. Considering the importance of gp120 amino (C1) and carboxyl (C5) segments in the association with e-gp41, we hypothesize that these regions are sufficient to maintain e-gp41 in a prefusion state. Based on the available gp120 atomic structure, we designed several truncated gp140 variants by including the C1 and C5 regions of gp120 in a gp41 ectodomain fragment. After iterative cycles of protein design, expression and characterization, we obtained a variant truncated at Lys(665) that stably folds as an elongated trimer under physiologic conditions. Several independent biochemical/biophysical analyses strongly suggest that this mini-Env adopts a prefusion e-gp41 configuration that is strikingly distinct from the postfusion trimer-of-hairpin structure. Interestingly, this prefusion mini-Env, lacking the fragment containing the 2F5/4E10 neutralizing monoclonal antibody binding sites, displays no detectable HIV-neutralizing epitopes when employed as an immunogen in rabbits. The result of this immunogenicity study has important implications for HIV-1 vaccine design efforts. Moreover, this engineered mini-Env protein should facilitate three-dimensional structural studies of the prefusion e-gp41 and serve to guide future attempts at pharmacologic and immunologic intervention of HIV-1. PMID- 15833741 TI - Nuclear-cytoplasmic shuttling of a RING-IBR protein RBCK1 and its functional interaction with nuclear body proteins. AB - The intracellular localization of a RING-IBR protein, RBCK1, possessing DNA binding and transcriptional activities, has been investigated. The endogenous RBCK1 was found in both the cytoplasm and nucleus. Particularly in the nucleus, it was localized in the granular structures, most likely nuclear bodies. In contrast, the over-expressed RBCK1 was detected exclusively in the cytoplasm. When the cells were treated with leptomycin B, the over-expressed RBCK1 accumulated in the nuclear bodies. These results suggest that RBCK1 possesses the signal sequences responsible for the nuclearcytoplasmic translocation. Mutational analysis of RBCK1 has indicated that an N-terminal region containing Leu-142 and Leu-145 and a C-terminal one containing the RING-IBR domain serve as the nuclear export and localization signals, respectively. Thus, RBCK1 is a transcription factor dynamically shuttling between cytoplasm and nucleus. Furthermore, RBCK1 was found to interact with nuclear body proteins, CREB-binding protein (CBP), and promyelocytic leukemia protein (PML). Coexpression of RBCK1 with CBP significantly enhanced the transcriptional activity of RBCK1. Although PML per se showed no effect on the transcriptional activity of RBCK1, the CBP-enhanced activity was repressed by coexpression with PML, presumably through the interaction of PML and CBP. Taken together, our data demonstrate that RBCK1 is involved in transcriptional machinery in the nuclear bodies, and its transcriptional activity is regulated by nucleocytoplasmic shuttling. PMID- 15833742 TI - Rapid electron transfer between monomers when the cytochrome bc1 complex dimer is reduced through center N. AB - We have obtained evidence for electron transfer between cytochrome b subunits of the yeast bc(1) complex dimer by analyzing pre-steady state reduction of cytochrome b in the presence of center P inhibitors. The kinetics and extent of cytochrome b reduced by quinol in the presence of variable concentrations of antimycin decreased non-linearly and could only be fitted to a model in which electrons entering through one center N can equilibrate between the two cytochrome b subunits of the bc(1) complex dimer. The b(H) heme absorbance in a bc(1) complex inhibited at center P and preincubated with substoichiometric concentrations of antimycin showed a red shift upon the addition of substrate, which indicates that electrons from the uninhibited center N in one monomer are able to reach the b(H) heme at the antimycin-blocked site in the other. The extent of cytochrome b reduction by variable concentrations of menaquinol could only be fitted to a kinetic model that assumes electron equilibration between center N sites in the dimer. Kinetic simulations showed that non-rate-limiting electron equilibration between the two b(H) hemes in the dimer through the two b(L) hemes is possible upon reduction through one center N despite the thermodynamically unfavorable b(H) to b(L) electron transfer step. We propose that electron transfer between cytochrome b subunits minimizes the formation of semiquinone-ferrocytochrome b(H) complexes at center N and favors ubiquinol oxidation at center P by increasing the amount of oxidized cytochrome b. PMID- 15833743 TI - Tpl2/cot signals activate ERK, JNK, and NF-kappaB in a cell-type and stimulus specific manner. AB - Macrophages and B-cells from Tpl2 knock-out mice exhibit a restricted defect in lipopolysaccharide and death receptor signaling that is limited to the activation of ERK. Here we show that Tpl2-/- MEFs exhibit defects in ERK, JNK, and NF-kappaB activation, or ERK activation only when stimulated with tumor necrosis factor alpha (TNF-alpha) or interleukin-1beta, respectively. In addition, we show that the activation of Tpl2 by TNF-alpha depends on signals transduced by both TRAF2 and RIP1. Activated Tpl2 phosphorylates MKK4/SEK1 upstream of JNK and stimulates NF-kappaB DNA binding and transcriptional activity by mechanisms that are independent of the nuclear translocation of p50 and p65. Tpl2-transduced TNF alpha signals instead promote the phosphorylation of p65 at Ser276 and modulate the spectrum of proteins associated with p65. Phosphorylation stimulates the transcriptional activity of NF-kappaB but does not affect its ability to bind DNA, which may be affected by the composition of the nuclear NF-kappaB complexes. These data confirm that defects caused by a single mutation may be cell-type and signal-specific and delineate the role of Tpl2 in the transduction of TNF-alpha signals that activate JNK and NF-kappaB in MEFs. PMID- 15833744 TI - Conkunitzin-S1 is the first member of a new Kunitz-type neurotoxin family. Structural and functional characterization. AB - Conkunitzin-S1 (Conk-S1) is a 60-residue neurotoxin from the venom of the cone snail Conus striatus that interacts with voltage-gated potassium channels. Conk S1 shares sequence homology with Kunitz-type proteins but contains only two out of the three highly conserved cysteine bridges, which are typically found in these small, basic protein modules. In this study the three-dimensional structure of Conk-S1 has been solved by multidimensional NMR spectroscopy. The solution structure of recombinant Conk-S1 shows that a Kunitz fold is present, even though one of the highly conserved disulfide cross-links is missing. Introduction of a third, homologous disulfide bond into Conk-S1 results in a functional toxin with similar affinity for Shaker potassium channels. The affinity of Conk-S1 can be enhanced by a pore mutation within the Shaker channel pore indicating an interaction of Conk-S1 with the vestibule of potassium channels. PMID- 15833745 TI - Monoclonal antibody epitope mapping describes tailspike beta-helix folding and aggregation intermediates. AB - There is growing interest in understanding how the cellular environment affects protein folding mechanisms, but most spectroscopic methods for monitoring folding in vitro are unsuitable for experiments in vivo or in other complex mixtures. Monoclonal antibody binding represents a sensitive structural probe that can be detected against the background of other cellular components. A panel of antibodies has been raised against Salmonella typhimurium phage P22 tailspike. In this report, nine alpha-tailspike antibody binding epitopes were characterized by measuring the binding of these monoclonal antibodies to tailspike variants bearing surface point mutations. These results reveal that the antibody epitopes are distributed throughout the tailspike structure, with several clustered in the central parallel beta-helix domain. The ability of each antibody to distinguish between tailspike conformational states was assessed by measuring antibody binding to tailspike in vitro refolding intermediates. Interestingly, the binding of all but one of the nine antibodies is sensitive to the tailspike conformational state. Whereas several antibodies bind preferentially to the tailspike native structure, the structural features that comprise the binding epitopes form with different rates. In addition, two antibodies preferentially recognize early refolding intermediates. Combined with the epitope mapping, these results indicate portions of the beta-helix form early during refolding, perhaps serving as a scaffold for the formation of additional structure. Finally, three of the antibodies show enhanced binding to non-native, potentially aggregation prone tailspike conformations. The refolding results indicate these non-native conformations form early during the refolding reaction, long before the appearance of native tailspike. PMID- 15833746 TI - Presenilin/gamma-secretase-mediated cleavage of the voltage-gated sodium channel beta2-subunit regulates cell adhesion and migration. AB - The voltage-gated sodium channel beta2-subunit (beta2) is a member of the IgCAM superfamily and serves as both an adhesion molecule and an auxiliary subunit of the voltage-gated sodium channel. Here we found that beta2 undergoes ectodomain shedding followed by presenilin (PS)-dependent gamma-secretase-mediated cleavage. 12-O-Tetradecanoylphorbol-13-acetate treatment or expression of an alpha secretase enzyme, ADAM10, resulted in ectodomain cleavage of beta2 in Chinese hamster ovary cells. Subsequent cleavage of the remaining 15-kDa C-terminal fragment (beta2-CTF) was independently inhibited by three specific gamma secretase inhibitors, expression of the dominant negative form of PS1, and in PS1/PS2 knock-out cells. gamma-Secretase inhibitor treatment also increased endogenous beta2-CTF levels in neuroblastoma cells and mouse primary neuronal cultures. In a cell-free gamma-secretase assay, we detected gamma-secretase activity-dependent generation of a 12 kDa beta2 intracellular domain (ICD), which was loosely associated with the membrane fraction. To assess the functional role of beta2 processing by gamma-secretase, we tested whether N-[N-(3,5 difluorophenylacetyl-l-alanyl)]-S-phenylglycine t-butylester (DAPT), a specific gamma-secretase inhibitor, would alter beta2-mediated cell adhesion and migration. We found that DAPT inhibited cell-cell aggregation and migration in a wound healing assay carried out with Chinese hamster ovary cells expressing beta2. DAPT also reduced migration of neuroblastoma cells in a modified Boyden chamber assay. Since DAPT treatment resulted in increased beta2-CTF levels, we also tested whether beta2-CTFs or beta2-ICDs would directly affect cell migration by overexpressing recombinant proteins. Interestingly, elevated levels of beta2 CTFs, but not ICDs, also blocked cell migration by 81 to 93%. Together, our findings show for the first time that beta2 is a PS/gamma-secretase substrate and gamma-secretase mediated cleavage of beta2-CTF is required for cell-cell adhesion and migration of beta2-expressing cells. PMID- 15833747 TI - C5L2 is a functional receptor for acylation-stimulating protein. AB - C5L2 binds acylation-stimulating protein (ASP) with high affinity and is expressed in ASP-responsive cells. Functionality of C5L2 has not yet been demonstrated. Here we show that C5L2 is expressed in human subcutaneous and omental adipose tissue in both preadipocytes and adipocytes. In mice, C5L2 is expressed in all adipose tissues, at levels comparable with other tissues. Stable transfection of human C5L2 cDNA into HEK293 cells results in ASP stimulation of triglyceride synthesis (TGS) (193 +/- 33%, 5 microM ASP, p < 0.001, where basal = 100%) and glucose transport (168 +/- 21%, 10 microM ASP, p < 0.001). C3a similarly stimulates TGS (163 +/- 12%, p < 0.001), but C5a and C5a des-Arg have no effect. The ASP mechanism is to increase Vmax of glucose transport (149%) and triglyceride (TG) synthesis activity (165%) through increased diacylglycerolacyltransferase activity (200%). Antisense oligonucleotide down regulation of C5L2 in human skin fibroblasts decreases cell surface C5L2 (down to 54 +/- 4% of control, p < 0.001, comparable with nonimmune background). ASP response is coordinately lost (basal TGS = 14.6 +/- 1.6, with ASP = 21.0 +/- 1.4 (144%), with ASP + oligonucleotides = 11.0 +/- 0.8 pmol of TG/mg of cell protein, p < 0.001). In mouse 3T3-L1 preadipocytes, antisense oligonucleotides decrease C5L2 expression to 69.5 +/- 0.5% of control, p < 0.001 (comparable with nonimmune) with a loss of ASP stimulation (basal TGS = 22.4 +/- 2.9, with ASP = 39.6 +/- 8.8 (177%), with ASP + oligonucleotides = 25.3 +/- 3.0 pmol of TG/mg of cell protein, p < 0.001). C5L2 down-regulation and decreased ASP response correlate (r = 0.761, p < 0.0001 for HSF and r = 0.451, p < 0.05 for 3T3-L1). In HEK-hC5L2 expressing fluorescently tagged beta-arrestin, ASP induced beta arrestin translocation to the plasma membrane and formation of endocytic complexes concurrently with increased phosphorylation of C5L2. This is the first demonstration that C5L2 is a functional receptor, mediating ASP triglyceride stimulation. PMID- 15833748 TI - Changes in atopy over a quarter of a century, based on cross sectional data at three time periods. PMID- 15833749 TI - Use of waist circumference to predict insulin resistance: retrospective study. PMID- 15833750 TI - Was Rodney Ledward a statistical outlier? Retrospective analysis using routine hospital data to identify gynaecologists' performance. AB - OBJECTIVES: To investigate whether routinely collected data from hospital episode statistics could be used to identify the gynaecologist Rodney Ledward, who was suspended in 1966 and was the subject of the Ritchie inquiry into quality and practice within the NHS. DESIGN: A mixed scanning approach was used to identify seven variables from hospital episode statistics that were likely to be associated with potentially poor performance. A blinded multivariate analysis was undertaken to determine the distance (known as the Mahalanobis distance) in the seven indicator multidimensional space that each consultant was from the average consultant in each year. The change in Mahalanobis distance over time was also investigated by using a mixed effects model. SETTING: NHS hospital trusts in two English regions, in the five years from 1991-2 to 1995-6. Population Gynaecology consultants (n = 143) and their hospital episode statistics data. MAIN OUTCOME MEASURE: Whether Ledward was a statistical outlier at the 95% level. RESULTS: The proportion of consultants who were outliers in any one year (at the 95% significance level) ranged from 9% to 20%. Ledward appeared as an outlier in three of the five years. Our mixed effects (multi-year) model identified nine high outlier consultants, including Ledward. CONCLUSION: It was possible to identify Ledward as an outlier by using hospital episode statistics data. Although our method found other outlier consultants, we strongly caution that these outliers should not be overinterpreted as indicative of "poor" performance. Instead, a scientific search for a credible explanation should be undertaken, but this was outside the remit of our study. The set of indicators used means that cancer specialists, for example, are likely to have high values for several indicators, and the approach needs to be refined to deal with case mix variation. Even after allowing for that, the interpretation of outlier status is still as yet unclear. Further prospective evaluation of our method is warranted, but our overall approach may be potentially useful in other settings, especially where performance entails several indicator variables. PMID- 15833751 TI - Reliability of orthostatic responses in healthy men aged between 65 and 75 years. AB - The purpose of this study was to investigate the short-, medium- and long-term reproducibility of cardiovascular responses during 90 degrees head-up tilt (HUT) in healthy older men. Twenty-eight healthy male subjects aged 69 (95% confidence intervals, 68-70) years participated in the study. Eight subjects underwent duplicate 90 degrees HUT tests on consecutive days, while 20 subjects underwent four 90 degrees HUT tests performed at baseline, and after 1 week, 1 month and 1 year. Following a 20-min supine resting period, each subject was rapidly tilted to the upright vertical position (90 degrees HUT) and remained in that position for 15 min. Beat-by-beat recordings of mean (MAP), systolic (SBP) and diastolic (DBP) pressures were made via Finapres, while heart rate (HR) was monitored continuously from an electrocardiogram. No significant test-retest differences (P > 0.05) were observed for the changes in HR, MAP, SBP or DBP during 90 degrees HUT. These measurements demonstrated high reproducibility (intraclass correlation coefficient, r = 0.91-0.99, P < 0.05). The supine resting and tilted HR, MAP, SBP and DBP over the 1-week, 1-month and 1-year period were not significantly different (P > 0.05) from baseline, and demonstrated high reproducibility (intraclass correlation coefficient, r = 0.82-0.98, P < 0.05). The results of this study demonstrate that in healthy older men, cardiovascular responses during orthostasis are highly reproducible, and this reproducibility is maintained over a 12-month period. These findings demonstrate that the 90 degrees HUT test offers a reproducible method of monitoring longitudinal orthostatic responses in healthy older men. PMID- 15833752 TI - Alterations in mouse cardiac proteome after in vivo myocardial infarction: permanent ischaemia versus ischaemia-reperfusion. AB - Mice are increasingly used to study the early molecular mechanisms inducing injury to the heart following myocardial infarction. To date, two-dimensional gel electrophoresis combined with mass spectrometry has not been applied to identify changes in protein expression in myocardial tissue of mice subjected in vivo to permanent ischaemia (PI) or ischaemia-reperfusion (IR). In the PI group, ischaemia was induced for 210 min by ligation of the left anterior descending coronary artery while in the IR group, ischaemia was maintained for 30 min and reperfusion was allowed for 180 min. In both groups, the area of the left ventricle at risk was processed for 2-dimensional gel electrophoresis. By comparing protein density changes in cytosolic as well as membrane fractions, we found a total of 32 protein spots that were differentially expressed. Twenty spots changed in expression level after PI alone, four spots after IR alone, and eight spots changed in both models. Identified proteins with MALDI TOF-TOF and LC MS/MS can be classified into functional groups of anticoagulant proteins, structural proteins, inflammatory-related proteins, transcription- and translation-related proteins, heat shock proteins (HSPs), metabolism-related proteins and miscellaneous. A remarkable finding was the IR-specific translocation of annexins (A3 and A5) from the cytosolic to the membrane compartment, a phenomenon that was verified by Western blotting. Four proteins were changed in expression level at multiple spot locations, characterized by a difference in isoelectric point. In the case of cardiac troponin T and HSP-20, these changes were also dependent on the model. In addition, one spot for the proteins adenylate kinase 1, cardiac troponin T and HSP-20 was uniquely present in the IR and/or PI groups and not in the respective sham groups. The specific alterations in protein expression that took place after PI and IR may stimulate the search for new tools to diagnoze myocardial infarction and to characterize specific pathology-related changes in protein expression. PMID- 15833753 TI - Influence of elevated renin substrate on angiotensin II and arterial blood pressure in conscious mice. AB - The present experiments were performed to determine the influence of intravenous administration of renin substrate on plasma angiotensin II levels and mean arterial blood pressure in conscious C57BL/6J mice. Mice with chronic indwelling femoral arterial and venous catheters were acutely or chronically administered intravenous doses of a synthetic peptide corresponding to the 14 amino acids on the N-terminal of angiotensinogen. A dose-dependent increase in arterial blood pressure was observed as the intravenous bolus dose of the renin substrate was increased from 0.18 to 180 nmol kg(-1) with a maximal increase in pressure of 40 +/- 3 mmHg achieved following administration of the 18 nmol kg(-1) bolus (n = 11). Additional experiments demonstrated that a sustained intravenous infusion of the renin substrate led to a long-term increase in arterial blood pressure. The continuous infusion of renin substrate at 0.05 nmol kg(-1) min(-1) for 3 days did not alter arterial blood pressure from the control level of 119 +/- 5 mmHg (n = 5); however, arterial blood pressure significantly increased to 129 +/- 6 mmHg with an infusion rate of 0.5 nmol kg(-1) min(-1) and further increased to 141 +/- 3 mmHg when the renin substrate infusion was increased to 5.0 nmol kg(-1) min( 1). Finally, the infusion of renin substrate at 5.0 nmol kg(-1) min(-1) resulted in a significant increase in plasma angiotensin II concentration from 34 +/- 6 pg ml(-1) in vehicle-infused mice to 288 +/- 109 pg ml(-1). These results demonstrate that modulation of the circulating level of angiotensinogen can alter the plasma angiotensin II level and arterial blood pressure in normal animals. PMID- 15833754 TI - Body maintenance and repair: how food and exercise keep the musculoskeletal system in good shape. AB - This article provides a personal view of how feeding and exercise acutely modify protein metabolism of human skeletal muscle, with discussion of the anabolic signalling mechanisms involved and some new findings on the metabolism of the turnover of collagen, tendon and bone. PMID- 15833755 TI - Depolarization pattern of ventricular epicardium in two-kidney one-clip hypertensive rats. AB - The present study is the first attempt to examine the effect of left ventricular hypertrophy (LVH) on the excitation pattern of the ventricular epicardium in experimental hypertensive rats. The left renal artery was clipped in Wistar rats (n = 8; 6-8 months old; weight, 174-295 g) to produce two-kidney one-clip (2K1C) hypertension. After 4 weeks, blood pressure was measured, and epicardial potential mapping was performed under sinus rhythm from 64 unipolar electrodes regularly distributed over the ventricular epicardium. Systolic blood pressure was approximately 40% higher in the rats with a clipped renal artery (162 +/- 14 mmHg, mean +/- s.d.) than in the normotensive rats (115 +/- 3 mmHg). LVH (approximately 23% increase in the ratio of the left ventricular weight to the body weight, P < 0.05) was observed in the 2K1C hypertensive rats. The depolarization pattern of the ventricular epicardium in the normotensive rats was similar to that in the rats with 2K1C hypertensive LVH. The duration of ventricular epicardial activation was shown to increase (approximately 35%, P < 0.05) in the hypertensive rats as compared to the normotensive animals. This study provides an explanation for alterations of the body surface potential distribution in hypertensive patients with LVH. PMID- 15833756 TI - High intensity exercise increases expression of matrix metalloproteinases in fast skeletal muscle fibres. AB - Metalloproteinases (MMPs) are proteolytic enzymes that function in the extracellular matrix to degrade connective tissues. While it is clear that exercise-induced injury in skeletal muscle promotes increased expression of MMPs, the relationship between exercise intensity and expression of MMPs in muscles is unknown. These experiments tested the hypothesis that exercise-induced expression of matrix metalloproteinases (MMP-2 and MMP-9) is dose-dependent such that high intensity endurance exercise increases MMP expression whereas low-intensity endurance exercise will not promote MMP expression in skeletal muscles. Female rats (4 months old) completed 2 weeks of treadmill running at either low (18 m min(-1); approximately 50% maximum oxygen consumption rate ) or high intensity (32 m min(-1); approximately 70% ; up to 50 min day(-1)). Non-running, sedentary animals served as controls. Muscle mRNA and protein levels of MMP-2 and MMP-9 were assessed in gastrocnemius, quadriceps and soleus muscles by reverse transcriptase-polymerase chain reaction and Western blotting, respectively. Results indicate that exercise did not alter MMP-9 in any of these skeletal muscles. Further, our data reveal that low-intensity exercise did not alter the expression of MMP-2 in any of the muscles investigated. In contrast, high intensity exercise increased both mRNA and protein levels of MMP-2 in skeletal muscles containing a high percentage of fast type II fibres (i.e. gastronemius and superficial quadriceps). These results support the hypothesis that high intensity exercise is required to promote the expression of MMP-2 in skeletal muscles and that the influence of exercise on MMP-2 expression is dominant in muscles containing a high percentage of fast fibres. PMID- 15833757 TI - Inhibition of carbachol-evoked oscillatory currents by the NO donor sodium nitroprusside in guinea-pig ileal myocytes. AB - The effect of sodium nitroprusside (SNP) on carbachol (CCh)-evoked inward cationic current (Icat) oscillations in guinea-pig ileal longitudinal myocytes was investigated using the whole-cell patch-clamp technique and permeabilized longitudinal muscle strips. SNP (10 microm) completely inhibited I(cat) oscillations evoked by 1 microm CCh. 1H-(1,2,4) Oxadiazole [4,3-a] quinoxaline-1 one (ODQ; 1 microm) almost completely prevented the inhibitory effect of SNP on Icat oscillations. 8-Bromo-guanosine 3',5'-cyclic monophosphate (8-Br-cGMP; 30 microm) in the pipette solution completely abolished Icat oscillations. However, a pipette solution containing Rp-8-Br-cGMP (30 microm) almost completely abolished the inhibitory effect of SNP on Icat oscillations. When the intracellular calcium concentration ([Ca2+]i) was held at a resting level using BAPTA (10 mm) and Ca2+ (4.6 microm) in the pipette solution, CCh (1 microm) evoked only the sustained component of Icat without any oscillations and SNP did not affect the current. A high concentration of inositol 1,4,5-trisphosphate (IP3; 30 microm) in the patch pipette solutions significantly reduced the inhibitory effect of SNP (10 microm) on Icat oscillations. SNP significantly inhibited the Ca2+ release evoked by either CCh or IP3 but not by caffeine in permeabilized preparations of longitudinal muscle strips. These results suggest that the inhibitory effects of SNP on Icat oscillations are mediated, in part, by functional modulation of the IP3 receptor, and not by the inhibition of cationic channels themselves or by muscarinic receptors in the plasma membrane. This inhibition seems to be mediated by an increased cGMP concentration in a protein kinase G-dependent manner. PMID- 15833758 TI - Low pulse pressure is independently related to elevated natriuretic peptides and increased mortality in advanced chronic heart failure. AB - AIMS: An increased pulse pressure (PP) has consistently predicted increased cardiovascular morbidity and mortality in cardiovascular risk patients and mild chronic heart failure (CHF). In contrast, a decreased PP was related to increased mortality in patients with acute decompensated heart failure. However, the predictive value of PP in patients with advanced CHF is not known. METHODS AND RESULTS: PP was analysed for its effect on mortality, adjusting for other modifiers of risk, using Cox proportional hazards regression analysis of data collected from 1901 patients with New York Heart Association Class III or IV CHF (mean age 65 and mean ejection fraction 0.26). Natriuretic peptides were measured in a subgroup. Multivariable Cox-regression analysis demonstrated that lower PP was associated with an increased mortality [hazard ratio (HR) 0.91 per 10 mmHg; 0.93-0.99], independent of mean arterial pressure (MAP) and other well known prognostic markers. In patients with a PP below the median value of 45 mmHg, PP was a stronger predictor of mortality than MAP (HR for PP 0.80 per 10 mmHg; 0.64 0.99). In patients with a PP above the median value of 45 mmHg, MAP was a stronger predictor of mortality than PP (HR for MAP 0.83 per 10 mmHg increase; 0.72-0.95). In addition, lower PP was independently related to increased atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP). CONCLUSION: In patients with advanced CHF, low PP is an independent predictor of mortality. In addition, low PP was related to increased levels of ANP and BNP. PMID- 15833759 TI - Evident, but poorly defined: left ventricular hypertrabeculation/noncompaction and its diagnostic criteria. PMID- 15833760 TI - Relationship between reduced elasticity of extracardiac vessels and left main stem coronary artery disease. PMID- 15833761 TI - Myocardial infarction increases ACE2 expression in rat and humans. PMID- 15833762 TI - Role of CXCR2 in cigarette smoke-induced lung inflammation. AB - It has been hypothesized that the destruction of lung tissue observed in smokers with chronic obstructive pulmonary disease and emphysema is mediated by neutrophils recruited to the lungs by smoke exposure. This study investigated the role of the chemokine receptor CXCR2 in mediating neutrophilic inflammation in the lungs of mice acutely exposed to cigarette smoke. Exposure to dilute mainstream cigarette smoke for 1 h, twice per day for 3 days, induced acute inflammation in the lungs of C57BL/6 mice, with increased neutrophils and the neutrophil chemotactic CXC chemokines macrophage inflammatory protein (MIP)-2 and KC. Treatment with SCH-N, an orally active small molecule inhibitor of CXCR2, reduced the influx of neutrophils into the bronchoalveolar lavage (BAL) fluid. Histological changes were seen, with drug treatment reducing perivascular inflammation and the number of tissue neutrophils. beta-Glucuronidase activity was reduced in the BAL fluid of mice treated with SCH-N, indicating that the reduction in neutrophils was associated with a reduction in tissue damaging enzymes. Interestingly, whereas MIP-2 and KC were significantly elevated in the BAL fluid of smoke exposed mice, they were further elevated in mice exposed to smoke and treated with drug. The increase in MIP-2 and KC with drug treatment may be due to the decrease in lung neutrophils that either are not present to bind these chemokines or fail to provide a feedback signal to other cells producing these chemokines. Overall, these results demonstrate that inhibiting CXCR2 reduces neutrophilic inflammation and associated lung tissue damage due to acute cigarette smoke exposure. PMID- 15833763 TI - rhVEGF treatment preserves pulmonary vascular reactivity and structure in an experimental model of pulmonary hypertension in fetal sheep. AB - We have previously shown that lung VEGF expression is decreased in a fetal lamb model of PPHN and that VEGF165 inhibition causes severe pulmonary hypertension in fetal lambs. Therefore, we hypothesized that treatment with rhVEGF165 would preserve endothelium-dependent vasodilation and reduce the severity of pulmonary vascular remodeling in an experimental model of PPHN. We studied the effects of daily intrapulmonary infusions of rhVEGF after partial ligation of the ductus arteriosus (DA). We performed surgery in 24 late-gestation fetal lambs and placed catheters in the main pulmonary artery, left atrium, and aorta for pressure measurements and in the left pulmonary artery for drug infusions. A pressure transducer was placed around the LPA to measure blood flow to the left lung (Qp), and the DA was surgically constricted to induce pulmonary hypertension. rhVEGF165 or vehicle was infused for 7 or 14 days. ACh or 8-BrcGMP was infused on days 2 and 13 to assess endothelium-dependent and -independent vasodilation, respectively. ACh-induced vasodilation was reduced in PPHN lambs after 14 days (change in Qp from baseline, 106% vs. 11%). In contrast, the response to ACh was preserved in lambs treated with rhVEGF (change in Qp, 94% vs. 90%). Pulmonary vasodilation to 8-BrcGMP was not altered in PPHN lambs or enhanced by VEGF treatment. rhVEGF treatment increased expression of lung eNOS protein and decreased pulmonary artery wall thickness by 34% vs. PPHN lambs. We conclude that VEGF165 preserves endothelium-dependent vasodilation, upregulates eNOS expression, and reduces the severity of pulmonary vascular remodeling in experimental PPHN. PMID- 15833764 TI - Toll-like receptor 4 antagonist (E5564) prevents the chronic airway response to inhaled lipopolysaccharide. AB - Although chronic inhalation of endotoxin or lipopolysaccharide (LPS) causes all of the classic features of asthma, including airway hyperreactivity, airway inflammation, and airway remodeling, the mechanisms involved in this process are not clearly understood. The objective of this study was to determine whether intratracheal treatment with LPS antagonist (E5564, a lipid A analog) prevented the development of chronic endotoxin-induced airway disease in a mouse model of environmental airway disease. Pretreatment with 10 and 100 microg of E5564 was found to inhibit the airway response (hyperreactivity and inflammation) for up to 48 h after the administration of the compound. Repeated dosing with 50 microg of E5564 intratracheally did not cause any measurable toxicity. Therefore, in a chronic experiment, mice were treated with either E5564 (50 microg) or vehicle three times weekly for 5 wk and simultaneously daily exposed to either LPS (4.65 +/- 0.30 microg/m3) or saline aerosol. E5564 was effective in decreasing the airway hyperreactivity to methacholine, the air space neutrophilia, the interleukin-6 in the lung lavage fluid, and the neutrophil infiltration of the airways 36 h after 5 wk of LPS inhalation. Less collagen deposition was observed in the airways of E5564-treated mice compared with vehicle-treated mice after a 4 wk recovery period. Our results indicate that E5564, a Toll-like receptor 4 antagonist, minimizes the physiological and biological effects of chronic LPS inhalation, suggesting a therapeutic role for competitive LPS antagonists in preventing or reducing endotoxin-induced environmental airway disease. PMID- 15833765 TI - Primary type II alveolar epithelial cells present microbial antigens to antigen specific CD4+ T cells. AB - Type II alveolar epithelial cells (AEC) can produce various antimicrobial and proinflammatory effector molecules. This, together with their abundance and strategic location, suggests a role in host defense against pulmonary pathogens. We report that murine type II AEC, like their human counterparts, express class II major histocompatibility complex (MHC). Using a murine model of pulmonary tuberculosis, we find that type II AEC become activated and have increased cell surface expression of class II MHC, CD54, and CD95 following infection. Type II AEC use the class II MHC pathway to process and present mycobacterial antigens to immune CD4+ T cells isolated from mice infected with Mycobacterium tuberculosis. Therefore, not only can type II AEC contribute to the pulmonary immunity by secreting chemokines that recruit inflammatory cells to the lung, but they can also serve as antigen-presenting cells. Although type II AEC are unlikely to prime naive T cells, their ability to present antigens to T cells demonstrates that they can participate in the effector phase of the immune response. This represents a novel role for type II AEC in the immunological response to pulmonary pathogens. PMID- 15833766 TI - Enhanced HIV-1 specific immune response by CpG ODN and HIV-1 immunogen-pulsed dendritic cells confers protection in the Trimera murine model of HIV-1 infection. AB - We have recently developed a novel small animal model for HIV-1 infection (Ayash Rashkovsky et al., http://www.fasebj.org/cgi/doi/10.1096/fj.04-3184fje; doi:10.1096/fj.04-3184fje). The mice were successfully infected with HIV-1 for 4 6 wk with different clades of either T- or M-tropic isolates. HIV-1 infection was accompanied by rapid loss of human CD4+ T cells, decrease in CD4/CD8 ratio, and increased T cell activation. HIV specific human humoral and cellular immune responses were observed in all HIV-1 infected animals. In the present study, HIV specific human immune responses, both humoral and cellular, were generated in noninfected Trimera mice, after their immunization with gp120-depleted HIV-1 antigen, presented by autologous human dendritic cells. Addition of CpG ODN to the antigen-pulsed DCs significantly enhanced (by 2- to 30-fold) the humoral and cellular HIV-1 specific immune responses. Only mice immunized with the HIV-1 immunogen and CpG were completely protected from infection with HIV-1 after challenge with high infection titers of the virus. This novel small animal model for HIV-1 infection may thus serve as an attractive platform for rapid testing of candidate HIV-1 vaccines and of adjuvants and may shorten the time needed for the development and final assessment of protective HIV-1 vaccines in human trials. PMID- 15833767 TI - A novel small animal model for HIV-1 infection. AB - Lethally irradiated normal BALB/c mice, reconstituted with murine SCID bone marrow and engrafted with human PBMC (Trimera mice), were used to establish a novel murine model for HIV-1 infection. The Trimera mice were successfully infected with different clades and primary isolates of T- and M-tropic HIV-1, with the infection persisting in the animals for 4-6 wk. Rapid loss of the human CD4+ T cells, decrease in CD4/CD8 ratio, and increased T cell activation accompanied the viral infection. All HIV-1 infected animals were able to generate both primary and secondary immune responses, including HIV specific human humoral and cellular responses. In addition to testing the efficacy of new antiviral compounds, this new murine HIV-1 model may be used for studying host-virus interactions and, most importantly, for screening and developing potential HIV-1 protective vaccines and adjuvants (Ayash-Rashkovsky et al., http://www.fasebj.org/cgi/doi/10.1096/fj.04-3185fje; doi:10.1096/fj.04-3185fje.). PMID- 15833769 TI - Inducing sensory stimulation in treatment of anorexia nervosa. PMID- 15833770 TI - Haemochromatosis and non-hepatic malignancy. PMID- 15833768 TI - Thrombospondin-1 up-regulates expression of cell adhesion molecules and promotes monocyte binding to endothelium. AB - Expression of cell adhesion molecules (CAM) responsible for leukocyte-endothelium interactions plays a crucial role in inflammation and atherogenesis. Up regulation of vascular CAM-1 (VCAM-1), intracellular CAM-1 (ICAM-1), and E selectin expression promotes monocyte recruitment to sites of injury and is considered to be a critical step in atherosclerotic plaque development. Factors that trigger this initial response are not well understood. As platelet activation not only promotes thrombosis but also early stages of atherogenesis, we considered the role of thrombospondin-1 (TSP-1), a matricellular protein released in abundance from activated platelets and accumulated in sites of vascular injury, as a regulator of CAM expression. TSP-1 induced expression of VCAM-1 and ICAM-1 on endothelium of various origins, which in turn, resulted in a significant increase of monocyte attachment. This effect could be mimicked by a peptide derived from the C-terminal domain of TSP-1 and known to interact with CD47 on the cell surface. The essential role of CD47 in the cellular responses to TSP-1 was demonstrated further using inhibitory antibodies and knockdown of CD47 with small interfering RNA. Furthermore, we demonstrated that secretion of endogenous TSP-1 and its interaction with CD47 on the cell surface mediates endothelial response to the major proinflammatory agent, tumor necrosis factor alpha (TNF-alpha). Taken together, this study identifies a novel mechanism regulating CAM expression and subsequent monocyte binding to endothelium, which might influence the development of anti-atherosclerosis therapeutic strategies. PMID- 15833771 TI - Bilateral adrenal haemorrhage and coeliac disease. PMID- 15833772 TI - Transient global amnesia or transient epileptic amnesia? PMID- 15833773 TI - Breaking the news. PMID- 15833774 TI - Preimplantation genetic diagnosis for neurofibromatosis type 1. AB - PGD is an alternative to prenatal diagnosis that circumvents therapeutic abortion. Diagnosis is carried out on single cells obtained from three-day-old embryos, and only those that are free of the disease under consideration are transferred to the mother. Neurofibromatosis type 1 (NF1) is a common neurocutaneous disorder, inherited as an autosomal dominant trait and caused by mutations in the NF1 gene. For some patients, PGD may be the only acceptable manner to ensure the birth of unaffected children. Because of the large number of known NF1 mutations, the development of mutation-specific single-cell protocols is impractical, labour-intensive and expensive. This paper discusses the development of five PGD protocols, three of which are based on multiplex PCR for microsatellite-markers linked to the NF1 gene. After a linkage study, the diagnosis can be established through the markers, thereby obviating the need to detect the mutation itself. This not only ensures the accurate diagnosis of the embryos, but also a prompt acceptance of PGD referrals since one protocol can be useful for several couples. In addition, two mutation-specific PCRs were developed for two couples where a marker-based protocol was not applicable. In total, 16 PGD cycles were carried out for six couples, which resulted in one ongoing pregnancy and the delivery of a healthy unaffected boy. PMID- 15833775 TI - Insulin and insulin-like growth factors inhibit and luteinizing hormone augments ovarian theca-interstitial cell apoptosis. AB - Theca-interstitial (T-I) cells play a fundamental role in the control of ovarian function. Steroidogenic activity and growth of the T-I cells are regulated by many paracrine and endocrine factors. However, little is known about the mechanisms controlling T-I death. In an in vitro model of apoptosis, purified rat T-I cells were cultured for 24 h with serum and subsequently for up to an additional 24 h with serum or in serum-free medium with or without insulin, insulin-like growth factors (IGF-I and IGF-II) and LH or 8-bromo-cyclic AMP (8Br cAMP). Apoptosis was identified by histological assessment of nuclear morphology and by detection of internucleosomal cleavage and quantified by determination of [alpha32P]-dideoxy-ATP 3'-end labeling of low molecular weight DNA. Serum withdrawal resulted in nuclear condensation and fragmentation into apoptotic bodies of T-I cells and led to pronounced DNA cleavage. Insulin (10 nM) protected T-I cells from apoptosis, reducing DNA fragmentation by 39 +/- 8% compared to serum-free controls. IGF-I (10 nM) and IGF-II (10 nM) had comparable antiapoptotic effects, decreasing DNA fragmentation by 55 +/- 9% and 37 +/- 14%, respectively. In contrast, LH (100 ng/ml) and 8Br-cAMP (1 mM) augmented the pro apoptotic effect of serum withdrawal, increasing DNA fragmentation by 85 +/- 55% and 72 +/- 42%, respectively. The antiapoptotic effects of insulin and IGFs and the pro-apoptotic effect of LH, acting via the cAMP system, may be important in the maintenance of T-I homeostasis. Moreover, excessive levels of insulin and free IGFs may lead to T-I cell hyperplasia characteristic of conditions such as polycystic ovary syndrome. PMID- 15833776 TI - Nasotracheal intubation: look before you leap. AB - Nasotracheal intubation is frequently used for airway management during maxillofacial surgery. Complications such as haemorrhage occur more frequently with this route of intubation than with the orotracheal route. This case report describes a male patient aged 51 yr who developed severe epistaxis after the tube had passed the nares. As an additional complication laryngoscopy and endotracheal intubation failed because of difficult airway (Cormack-Lehane grade 4). Attempts using an intubating laryngeal mask airway and a Bonfils intubating fibrescope did not succeed. The airway was finally managed by cricothyroidotomy. A modified sequence of nasotracheal intubation is proposed to avoid similar life-threatening complications. PMID- 15833777 TI - Comparison of the respiratory effects of intravenous buprenorphine and fentanyl in humans and rats. AB - BACKGROUND: There is evidence from animal studies suggesting the existence of a ceiling effect for buprenorphine-induced respiratory depression. To study whether an apparent ceiling effect exists for respiratory depression induced by buprenorphine, we compared the respiratory effects of buprenorphine and fentanyl in humans and rats. METHODS: In healthy volunteers, the opioids were infused i.v. over 90 s and measurements of minute ventilation at a fixed end-tidal PCO2 of 7 kPa were obtained for 7 h. Buprenorphine doses were 0.7, 1.4, 4.3 and 8.6 microg kg(-1) (n=20 subjects) and fentanyl doses 1.1, 2.1, 2.9, 4.3 and 7.1 microg kg( 1) (n=21). Seven subjects received placebo. In rats, both opioids were infused i.v. over 20 min, and arterial PCO2 was measured 5, 10, 15 and 20 min after the start of fentanyl infusion and 30, 150, 270 and 390 min after the start of buprenorphine infusion. Doses tested were buprenorphine 0, 100, 300, 1000 and 3000 microg kg(-1) and fentanyl 0, 50, 68 and 90 microg kg(-1). RESULTS: In humans, fentanyl produced a dose-dependent depression of minute ventilation with apnoea at doses > or = 2.9 microg kg(-1); buprenorphine caused depression of minute ventilation which levelled off at doses > or = 3.0 microg kg(-1) to about 50% of baseline. In rats, the relationship of arterial PCO2 and fentanyl dose was linear, with maximum respiratory depression at 20 min (maximum PaCO2 8.0 kPa). Irrespective of the time at which measurements were obtained, buprenorphine showed a non-linear effect on PaCO2, with a ceiling effect at doses > 1.4 microg kg(-1). The effect on PaCO2 was modest (maximum value measured, 5.5 kPa). CONCLUSIONS: Our data confirm a ceiling effect of buprenorphine but not fentanyl with respect to respiratory depression. PMID- 15833778 TI - Predicting postoperative analgesia outcomes: NNT league tables or procedure specific evidence? AB - Number needed to treat (NNT) values have been recommended and used to assess efficacy of analgesics for acute pain management. However, the data analysed come from a variety of procedures, which may potentially hinder the interpretation of the NNT value for specific procedures. We reanalysed available NNT data with acetaminophen in relation to the magnitude of surgical injury. Acetaminophen was less effective for pain relief after orthopaedic procedures than after dental procedures. The relative risk ratio for more than 50% pain relief, compared with placebo, was only 1.87 compared with 3.77 (P<0.05). Although NNT can give a valuable overview of efficacy, this concept is not necessarily applicable to all types of surgery. We suggest that estimates of NNT should be related to specific surgical procedures. PMID- 15833779 TI - Effect of three different anaesthetic agents on the postoperative production of cardiac troponin T in paediatric cardiac surgery. AB - BACKGROUND: Paediatric cardiac surgery is associated with some degree of myocardial injury. Ischaemic preconditioning (IP) has been investigated widely in the adult population. Volatile agents have been shown to simulate IP providing extra protection to the myocardium during adult cardiopulmonary bypass (CPB) while propofol seems to act through different mechanisms. IP has not been investigated in the paediatric population to the same extent. Cardiac troponin T (cTnT) is a reliable marker of myocardial injury in neonates and children. We have investigated the relationship between three anaesthetic agents, midazolam, propofol, and sevoflurane, and postoperative production of cTnT. METHODS: Ninety patients undergoing repair of congenital heart defect with CPB were investigated in a prospective randomized study. cTnT was measured four times during the first 24 h following admission to the paediatric intensive care unit. Other variables measured included arterial blood gases, lactate, fluid balance, use of inotropic drugs, PaO2/FiO2 ratio and ventilator hours. RESULTS: cTnT was elevated in all three groups throughout the study period. The differences between the three groups were not statistically significant. Eight hours after admission to the intensive care unit cTnT concentrations tended to be higher in the midazolam group [mean (95% confidence intervals)]; 2.7 (1.9-3.5) ng ml(-1). Patients receiving a propofol-based anaesthesia had similar concentrations 2.6 (1.7-3.5) ng ml(-1) while those receiving sevoflurane tended to have a lower cTnT production 1.7 (1.3-2.2) ng ml(-1). CONCLUSIONS: Midazolam, propofol, and sevoflurane appear to provide equal myocardial protection in paediatric cardiac surgery when using cTnT as a marker of myocardial damage. PMID- 15833780 TI - Comparison of propofol/remifentanil and sevoflurane/remifentanil for maintenance of anaesthesia for elective intracranial surgery. AB - BACKGROUND: Propofol and sevoflurane are suitable agents for maintenance of anaesthesia during neurosurgical procedures. We have prospectively compared these agents in combination with the short-acting opioid, remifentanil. METHODS: Fifty unpremedicated patients undergoing elective craniotomy received remifentanil 1 microg kg(-1) followed by an infusion commencing at 0.5 microg kg(-1) min(-1) reducing to 0.25 microg kg(-1) min(-1) after craniotomy. Anaesthesia was induced with propofol, and maintained with either a target-controlled infusion of propofol, minimum target 2 microg ml(-1) or sevoflurane, initial concentration 2%(ET). Episodes of mean arterial pressure (MAP) more than 100 mm Hg or less than 60 mm Hg for more than 1 min were defined as hypertensive or hypotensive events, respectively. A surgical assessment of operating conditions and times to spontaneous respiration, extubation, obey commands and eye opening were recorded. Drug acquisition costs were calculated. RESULTS: Twenty-four and twenty-six patients were assigned to propofol (Group P) and sevoflurane anaesthesia (Group S), respectively. The number of hypertensive events was comparable, whilst more hypotensive events were observed in Group S than in Group P (P=0.053, chi-squared test). As rescue therapy, more labetolol [45 (33) vs 76 (58) mg, P=0.073] and ephedrine [4.80 (2.21) vs 9.78 (5.59) mg, P=0.020] were used in Group S. Between group differences in recovery times were small and clinically unimportant. The combined hourly acquisition costs of hypnotic, analgesic, and vasoactive drugs appeared to be lower in patients maintained with sevoflurane than with propofol. CONCLUSION: Propofol/remifentanil and sevoflurane/remifentanil both provided satisfactory anaesthesia for intracranial surgery. PMID- 15833781 TI - Impact of age on both BIS values and EEG bispectrum during anaesthesia with sevoflurane in children. AB - BACKGROUND: The aim of this study was to evaluate the potential relationship between age, BIS (Aspect), and the EEG bispectrum during anaesthesia with sevoflurane. METHODS: BIS and raw EEG were recorded at a steady state of 1 MAC in 100 children, and during a decrease from 2 to 0.5 MAC in a sub-group of 29 children. The bispectrum of the EEG was estimated using MATLAB software. For analysis, the bispectrum was divided into 36 frequencies of coupling (P(i))--the MatBis. A multiple correspondence analysis (MCA) was used to establish an underlying structure of the pattern of each individual's MatBis at 1 MAC. Clustering of children into homogeneous groups was conducted by a hierarchical ascending classification (HAC). The level of statistical significance was set at 0.05. RESULTS: At 1 MAC, the BIS values for all children ranged from 20 to 74 (median 40). Projection of both age and BIS value recorded at 1 MAC onto the structured model of the MCA showed them to be distributed along the same axis, demonstrating that the different values of BIS obtained in younger or older children are mainly dependent on their MatBis. At 1 MAC, six homogeneous groups of children were obtained through the HAC. Groups 5 (30 months; range 23-49) and 6 (18 months; range 6-180) were the younger children and Group 1 (97 months; range 46-162) the older. Groups 5 and 6 had the highest median values of BIS (54; range 50-59) (55; range 26-74) and Group 1 the lowest values (29; range 22-37). CONCLUSION: The EEG bispectrum, as well as the BIS appeared to be strongly related to the age of children at 1 MAC sevoflurane. PMID- 15833782 TI - Antiemetic and analgesic-sparing effects of diphenhydramine added to morphine intravenous patient-controlled analgesia. AB - BACKGROUND: This study was designed to examine the analgesic and dose-related antiemetic efficacy of diphenhydramine-morphine mixture for intravenous patient controlled analgesia (PCA). METHODS: Healthy women, undergoing abdominal total hysterectomy were recruited to this double-blinded randomized placebo-controlled study. Patients were randomly allocated to one of three groups (n=40 each). In group 1, patients received saline at induction and morphine 1 mg ml(-1) alone for postoperative PCA. Patients in groups 2 and 3 received diphenhydramine 30 mg i.v. at induction and were given a 1.2:1 or a 4.8:1 ratio, respectively, of diphenhydramine-morphine mixture for postoperative PCA. RESULTS: A total of 112 patients completed the study. The incidence of postoperative nausea (31.6% vs 67.6%, P<0.01) and vomiting (15.8% vs 40.5%, <0.05) was significantly lower in group 3 than in group 1. Furthermore, the incidence of severe nausea was significantly lower in group 3 than in group 1 (2.6% vs 24.3%, P<0.05). The rescue antiemetic requirements were also significantly less in group 3 than in group 1 (5.3% vs 24.3%, P<0.05). However, there was no significant difference between group 2 and group 1 in any of the comparisons. Pain intensity, 24-h morphine consumption and diphenhydramine-related side-effects, such as sedation or dry mouth, did not differ among the three groups. CONCLUSION: An initial bolus of diphenhydramine 30 mg at anaesthetic induction followed by postoperative PCA with a 4.8:1, but not 1.2:1, diphenhydramine-morphine mixture provides an effective antiemetic efficacy without morphine-sparing effects. PMID- 15833783 TI - Dysprealbuminemic hyperthyroxinemia in a patient with hyperthyroid graves disease. AB - Rare mutant forms of circulating albumin and prealbumin [transthyretin (TTR)] have increased binding affinity for thyroxine (T4). Patients with these variant plasma proteins, as a result of inherited mutations or as a paraneoplastic phenomenon, typically present with increased serum total T4 and, by some assay methodologies, an increased free T4 as well. Although these individuals are, in fact, euthyroid, nonspecific symptoms may lead to inappropriate treatment for hyperthyroidism. We present a 34-year-old woman in whom a mutant form of TTR with increased T4 binding affinity and coexisting Graves disease was present. Subsequent 131I therapy led to development of postablative hypothyroidism, which was obscured by her higher serum free T4 concentration. Circulating thyroid binding globulin (TBG), albumin, and TTR concentrations were all within their respective reference limits. A T4-binding protein panel confirmed that TTR-bound T4 was significantly increased, whereas TBG- and albumin-bound T4 was normal, indicating that this patient had euthyroid dysprealbuminemic hyperthyroxinemia, which had been masked by the initial presentation of hyperthyroidism. These findings indicate that hypothyroidism can be masked by coexisting euthyroid dysprealbuminemic hyperthyroxinemia. PMID- 15833784 TI - Comparison of the unsaturated iron-binding capacity with transferrin saturation as a screening test to detect C282Y homozygotes for hemochromatosis in 101,168 participants in the hemochromatosis and iron overload screening (HEIRS) study. PMID- 15833785 TI - Fast and slow skeletal troponin I in serum from patients with various skeletal muscle disorders: a pilot study. AB - BACKGROUND: Detection of skeletal muscle injury is hampered by a lack of commercially available assays for serum markers specific for skeletal muscle; serum concentrations of skeletal troponin I (sTnI) could meet this need. Moreover, because sTnI exists in 2 isoforms, slow (ssTnI) and fast (fsTnI), corresponding to slow- and fast-twitch muscles, respectively, it could provide insight into differential injury/recovery of specific fiber types. The purpose of this study was to investigate whether the 2 isoforms of sTnI and their modified forms are present in the blood of patients with various skeletal muscle disorders. METHODS: Serial serum samples were obtained from 25 patients with various skeletal muscle injuries. Serum proteins were separated by a modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis protocol followed by Western blotting for sTnI with monoclonal antibodies specific to ssTnI and fsTnI. RESULTS: We observed (a) intact and, in some cases, degraded sTnI products; (b) evidence of posttranslational modifications in addition to proteolysis; and (c) differential detectability of both skeletal isoforms in the same patient. CONCLUSIONS: It is possible to monitor both sTnI isoforms; this could lead to the development of new diagnostic assays for skeletal muscle damage. PMID- 15833786 TI - Should verbal autopsy results for malaria be adjusted to improve validity? PMID- 15833787 TI - Case recruitment in genetic association studies: larger sample size or greater homogeneity? PMID- 15833788 TI - Fetal death sex ratios: a test of the economic stress hypothesis. AB - BACKGROUND: The ratio of male to female live births (i.e. the sex ratio) reportedly falls when populations suffer rare and extreme ambient stressors such as the collapse of national economies. This association has been attributed to the death of male fetuses and to reduced conception of males. We assess the validity of the first of these mechanisms by testing the hypothesis that the fetal death sex ratio varies positively over time with the unemployment rate. Using the unemployment rate also allows us to determine if ambient economic stressors less extreme than collapsing national economies affect the fetal death sex ratio. METHODS: We test our hypotheses by applying time-series methods to monthly counts of fetal deaths and the unemployment rate from the state of California beginning January 1989 and ending December 2001. The methods control for trends, seasonal cycles, and other forms of autocorrelation that could induce spurious associations. RESULTS: Results support the fetal death mechanism in that male fetal deaths increased above the values expected from female deaths and from history in months in which the unemployment rate also increased over its expected value. CONCLUSIONS: Our findings suggest that ambient stressors as common as increasing unemployment elevate the risk of fetal death among males. We discuss the social, economic, and health costs borne by parents and communities afflicted with these fetal deaths. PMID- 15833789 TI - Alcohol consumption and its relation to risk factors for cardiovascular disease in the north-west of Russia: the Arkhangelsk study. AB - BACKGROUND: To explore indicators and levels of alcohol consumption in a Russian population, and to elaborate these in relation to risk factors for cardiovascular disease. METHODS: A total of 1963 men and 1734 women, aged 18-75 years, consecutively recruited at their compulsory annual medical check-up at the Semashko outpatient clinic, Arkhangelsk, participated in a cross-sectional health survey. The survey comprised a physical examination, a six-page questionnaire on health and lifestyle, and blood tests. RESULTS: Gamma-glutamyltransferase (GGT) levels in both sexes were more than twice as high as found in comparable studies. Elevated GGT-levels were 4-5 times more frequent than found in Norwegian studies. Alcohol Use Disorder Identification Test (AUDIT) identified up to 75% of male workers and 47% of female workers as hazardous or harmful alcohol drinkers. The traditional risk factors for cardiovascular disease were significantly higher in subjects with a high level of GGT. CONCLUSION: The findings indicate an extremely high level of alcohol consumption in this population. Elevation in GGT was significantly associated with increased risk for cardiovascular disease. PMID- 15833790 TI - Commentary: Smoke pulls the blinds. PMID- 15833791 TI - Commentary: On the article by Lester Breslow on the origins and development of the IEA. PMID- 15833792 TI - Origins and development of the International Epidemiological Association. PMID- 15833793 TI - Commentary: Modelling multiple pathways to explain social inequalities in health and mortality. PMID- 15833794 TI - Towards the global elimination of brain damage due to iodine deficiency--the role of the International Council for Control of Iodine Deficiency Disorders. PMID- 15833795 TI - Why do women complain of vaginal discharge? A population survey of infectious and pyschosocial risk factors in a South Asian community. AB - BACKGROUND: Vaginal discharge is a common complaint, particularly among women in Asia. Although presumed to be caused by reproductive tract infections (RTIs), the association between the complaint and the presence of RTIs is weak. This study aimed to investigate the risk factors of the complaint of vaginal discharge. METHODS: We conducted a community-based survey of 3000 women aged 18-50 years, randomly sampled from a population in Goa, India. Women who gave informed consent were invited to participate in a structured interview, which elicited data on the primary outcome (the experience of current abnormal vaginal discharge) and psychosocial exposures: gender adversity; symptoms of somatoform disorders; and common mental disorders (CMD). All women were required to provide vaginal and/or urine samples for diagnosis of RTIs using gold standard laboratory tests. Risk factors were analysed using logistic regression with the binary outcome of the complaint of vaginal discharge. RESULTS: Of the 2494 women (83%) who agreed to participate, 14.5% complained of having an abnormal vaginal discharge. Stress was the most common causal attribution for the complaint. The final multivariate model found that high scores for CMD (OR 2.16, 1.4-3.2) and somatoform disorders (6.23, 4.0-9.7) and the use of an intrauterine contraceptive device (1.86, 1.0 3.4) were independently associated with the complaint. Low literacy (0.54, 0.4 0.8) and age >40 years (0.29, 0.2-0.4) were associated with a reduced risk. RTI were not associated with the complaint (1.24, 0.9-1.6). CONCLUSIONS: Psychosocial factors have the strongest association with the complaint of vaginal discharge. Syndromic management algorithms need refinement so that women with complaints that are non-infectious in aetiology are offered psychosocial interventions. PMID- 15833796 TI - Influence of short stature on the change in pulse pressure, systolic and diastolic blood pressure from age 36 to 53 years: an analysis using multilevel models. AB - BACKGROUND: Previous cross-sectional analyses of this cohort have shown that short height and leg length are associated with higher pulse pressure and systolic blood pressure in middle age. It is unclear how these adult measures of childhood growth influence the change in blood pressure as it increases with age. METHODS: Multilevel models were fitted to investigate associations between components of height and the change in blood pressure between 36, 43, and 53 years in a prospective national cohort of 1472 men and 1563 women followed-up since birth in 1946. RESULTS: Shorter height and leg length, but not trunk length, were associated with higher blood pressure, similarly in men and women. Longitudinal analyses showed that the effects of both height and leg length on pulse pressure and systolic blood pressure became significantly stronger with age. For example, the change in systolic blood pressure was found to be -0.021 mm Hg (95% confidence interval -0.029 to -0.013) per year lower for every centimetre increase in leg length (P < or = 0.001). In other words, the increase in systolic blood pressure over a 10 year period of a participant whose legs were 10 centimetres shorter was 2.1 mm Hg higher (P < or = 0.001), compared with a taller participant. Associations were independent of a number of potential confounders. CONCLUSIONS: These results support the hypothesis that short people may be more susceptible to the effects of ageing on the arterial tree. Childhood growth may contribute to the tracking of cardiovascular risk throughout life. PMID- 15833797 TI - Microdialysis separately monitors myocardial interstitial myoglobin during ischemia and reperfusion. AB - Direct monitoring of myoglobin efflux during ischemia and reperfusion has been limited because of inherent sample collection problems in the ischemic region. Recently, the cardiac dialysis technique has offered a powerful method for monitoring myocardial interstitial levels of low-molecular-weight compounds in the cardiac ischemic region. In the present study, we extended the molecular target to high-molecular-weight compounds by use of microdialysis probes with a high-molecular-mass cutoff and monitored myocardial interstitial myoglobin levels. A dialysis probe was implanted in the left ventricular free wall in anesthetized rabbits. The main coronary artery was occluded for 60 or 120 min. We examined the effects of myocardial ischemia and reperfusion on myocardial interstitial myoglobin levels. Interstitial myoglobin increased within 15 min of ischemia and continued to increase during 120 min of ischemia, whereas blood myoglobin increased at 45 min of ischemia. Lactate and myoglobin in the interstitial space increased during the same period. At 60 min of ischemia, reperfusion markedly accelerated interstitial myoglobin release. The interstitial myoglobin level was fivefold higher at 0-15 min of reperfusion than at 60-75 min of coronary occlusion. The dialysis technique permits earlier detection of myoglobin release and separately monitors myoglobin release during ischemia and reperfusion. Myocardial interstitial myoglobin levels can serve as an index of myocardial injury evoked by ischemia or reperfusion. PMID- 15833798 TI - Tempol therapy attenuates medial smooth muscle cell apoptosis and neointima formation after balloon catheter injury in carotid artery of diabetic rats. AB - Accumulating data support the hypothesis that reactive oxygen species (ROS) play a critical role in the vascular complications observed in diabetes. However, the mechanisms of ROS-mediated vascular complications in diabetes are not clear. We tested the hypothesis that ROS-mediated increase in proapoptotic factor Bax expression leads to medial smooth muscle cell (SMC) apoptosis that is associated with neointima formation. We used a fructose-rich diet for 4 wk to model Type 2 diabetes in rats. SOD mimetic membrane-permeable 4-hydroxy-2,2,6,6, tetramethylpiperidine-1-oxyl (Tempol, 1 mM) was administered in drinking water to scavenge superoxide starting 1 day before surgery and continued during the duration of the experiment. Vascular injury resulted in a significant increase in medial SMC apoptosis that was associated with neointima formation. The number of medial SMC positive for Bax immunostaining significantly increased in injured arteries compared with uninjured arteries. Superoxide scavenging by Tempol treatment inhibited both the Bax-positive index as well as the apoptotic index of medial SMC in response to vascular injury. Tempol treatment inhibited apoptotic loss of medial SMC, thus increasing their density in the injured arteries. These alterations in the media were associated with a marked decrease in neointima formation in injured arteries. We conclude that Bax expression may play an important role in vascular SMC apoptosis and, finally, that this regulatory mechanism is redox sensitive. PMID- 15833799 TI - Delayed adenosine A1 receptor preconditioning in rat myocardium is MAPK dependent but iNOS independent. AB - Adenosine A1 receptor delayed preconditioning (PC) against myocardial infarction has been well described; however, there have been limited investigations of the signaling mechanisms that mediate this phenomenon. In addition, there are multiple conflicting reports on the role of inducible nitric oxide synthase (iNOS) in mediating A1 late-phase PC. The purpose of this study was to determine the roles of the p38 and extracellular signal-regulated kinase (ERK) mitogen activated protein kinases (MAPKs) in in vivo delayed A1 receptor PC and whether this protection at the myocyte level is due to upregulation of iNOS. Myocardial infarct size was measured in open-chest anesthetized rats 24 h after treatment with vehicle or the adenosine A1 agonist 2-chloro-N6-cyclopentyladenosine (CCPA; 100 microg/kg ip). Additional rats receiving CCPA were pretreated with the p38 inhibitor SB-203580 (1 mg/kg ip) or the MAPK/ERK kinase (MEK) inhibitor PD-098059 (0.5 mg/kg ip). At 24 h after CCPA administration, a group of animals was given the iNOS inhibitor 1400 W 10 min before ischemia. Treatment with CCPA reduced infarct size from 48 +/- 2 to 28 +/- 2% of the area at risk, an effect that was blocked by both SB-203580 and PD-098059 but not 1400 W. Ventricular myocytes isolated 24 h after CCPA injection exhibited significantly reduced oxidative stress during H2O2 exposure compared with myocytes from vehicle-injected animals, and this effect was not blocked by the iNOS inhibitor 1400 W. Western blot analysis of whole heart and cardiac myocyte protein samples revealed no expression of iNOS 6 or 24 h after CCPA treatment. These results indicate that adenosine A1 receptor delayed PC in rats is mediated by MAPK-dependent mechanisms, but this phenomenon is not associated with the early or late expression of iNOS. PMID- 15833800 TI - Thrombin, TNF-alpha, and LPS exert overlapping but nonidentical effects on gene expression in endothelial cells and vascular smooth muscle cells. AB - Thrombin, TNF-alpha, and LPS have each been implicated in endothelial cell and vascular smooth muscle cell (VSMC) activation. We wanted to test the hypothesis that these three agonists display mediator and/or cell type-specific properties. The addition of thrombin to human pulmonary artery endothelial cells resulted in an upregulation of PDGF-A, tissue factor (TF), ICAM-1, and urokinase-type plasminogen activator (u-PA), whereas TNF-alpha and LPS failed to induce PDGF-A. These effects were mimicked by protease-activated receptor-1 activation. In VSMC, thrombin induced expression of TF and PDGF-A but failed to consistently induce ICAM-1 or u-PA expression. In contrast, TNF-alpha and LPS increased expression of all four genes in this cell type. Inhibitor studies in endothelial cells demonstrated a critical role for PKC in mediating thrombin, TNF-alpha, and LPS induction of ICAM-1, TF, and u-PA and for p38 MAPK in mediating thrombin, TNF alpha, and LPS induction of TF. Taken together, these results suggest that inflammatory mediators engage distinct signaling pathways and expression profiles in endothelial cells and VSMC. The data support the notion that endothelial cell activation is not an all-or-nothing phenomenon but rather is dependent on the nature of the extracellular mediator. PMID- 15833801 TI - Catecholamines augment collateral vessel growth and angiogenesis in hindlimb ischemia. AB - Catecholamine stimulation of alpha1-adrenoceptors exerts growth factor-like activity, mediated by generation of reactive oxygen species, on arterial smooth muscle cells and adventitial fibroblasts and contributes to hypertrophy and hyperplasia in models of vascular injury and disease. Adrenergic trophic activity also contributes to flow-mediated positive arterial remodeling by augmenting proliferation and leukocyte accumulation. To further examine this concept, we studied whether catecholamines contribute to collateral growth and angiogenesis in hindlimb insufficiency. Support for this hypothesis includes the above mentioned studies, evidence that ischemia augments norepinephrine release from sympathetic nerves, and proposed involvement of reactive oxygen species in angiogenesis and collateral growth. Mice deficient in catecholamine synthesis [by gene deletion of dopamine beta-hydroxylase (DBH-/-)] were studied. At 3 wk after femoral artery ligation, increases in adductor muscle perfusion were similar in DBH-/- and wild-type mice, whereas recovery of plantar perfusion and calf microsphere flow were attenuated, although not significantly. Preexisting collaterals in adductor of wild-type mice showed increases in lumen diameter (60%) and medial and adventitial thickness (57 and 119%, P < 0.05 here and below). Lumen diameter increased similarly in DBH-/- mice (52%); however, increases in medial and adventitial thicknesses were reduced (30 and 65%). Leukocyte accumulation in the adventitia/periadventitia of collaterals was 39% less in DBH-/- mice. Increased density of alpha-smooth muscle actin-positive vessels in wild-type adductor (45%) was inhibited in DBH-/- mice (2%). Although both groups experienced similar atrophy in the gastrocnemius (approximately 22%), the increase in capillary-to-muscle fiber ratio in wild-type mice (21%) was inhibited in DBH-/- mice (7%). These data suggest that catecholamines may contribute to collateral growth and angiogenesis in tissue ischemia. PMID- 15833802 TI - Effects of pulsation frequency and endothelial integrity on enhanced arterial transmural filtration produced by pulsatile pressure. AB - The role of the endothelium in regulating transmural fluid filtration into the artery wall under pulsatile pressure and the effects of changes in pulsatile frequency on filtration have received little attention. Previous experiments (Alberding JP, Baldwin AL, Barton JK, and Wiley E. Am J Physiol Heart Circ Physiol 286: H1827-H1835, 2004) demonstrated significantly increased filtration after initial onset of pulsatile pressure compared with that predicted by using parameters measured under steady pressure. To determine the role of the endothelium in this phenomenon, the following experiments were performed on five New Zealand White rabbits (anesthetized with 30 mg/kg pentobarbital sodium). One of each pair of carotid arteries was deendothelialized, and filtration measurements under steady and pulsatile pressure were compared with those made in intact vessels (Alberding JP, Baldwin AL, Barton JK, and Wiley E. Am J Physiol Heart Circ Physiol 286: H1827-H1835, 2004). To determine the effect of increasing pulsatile frequency on arterial filtration, transmural filtration was measured by using pulsatile pressure frequencies of 1 Hz, followed by 2 Hz, in another set of intact arteries (6 arteries and 3 animals). For deendothelialized vessels, the initial increase in filtration after onset of pulsatility was similar to that observed in intact vessels, but the subsequent reduction in filtration was less abrupt. When pulsatile frequency was increased from 1 to 2 Hz in intact arteries, an initial increase in filtration was observed, similar to that obtained after onset of pulsatile pressure subsequent to a steady pressure. The observed responses of arteries to pulsatile pressure, with and without endothelium, or undergoing a frequency change, suggest a dynamic role for the endothelium in regulating transvascular transport in vivo. PMID- 15833803 TI - Heterozygous knockout of neuregulin-1 gene in mice exacerbates doxorubicin induced heart failure. AB - Neuregulins and their erbB receptors are essential for cardiac development and postulated to be cardioprotective in the presence of injury in the postnatal heart. We tested the hypothesis that the development of doxorubicin-induced cardiotoxicity in vivo is more severe in mice with heterozygous knockout of the neuregulin-1 gene (NRG-1(+/-)) compared with wild-type mice (WT). Three-month old NRG-1(+/-) and WT mice were injected with a single dose of doxorubicin (20 mg/kg ip). Survival was analyzed by the Kaplan-Meier approach. Left ventricular (LV) function and signaling pathways were analyzed 4 days after treatment. Fifteen days after treatment, survival was significantly lower in doxorubicin-treated NRG 1(+/-) mice (NRG-1(+/-)-Dox) compared with doxorubicin-treated WT mice (WT-Dox) (15% vs. 33%, P < 0.01). LV mass was significantly lower in NRG-1(+/-)-Dox but not in WT-Dox compared with nontreated animals. LV systolic pressure and LV midwall fractional shortening were significantly lower in NRG-1(+/-)-Dox compared with WT-Dox mice. LV protein levels of NRG-1, erbB2, and erbB4 receptors were similar in WT-Dox and NRG-1(+/-)-Dox mice. However, levels of phosphorylated erbB2, Akt, and ERK-1/2 were significantly decreased in NRG-1(+/-)-Dox compared with WT-Dox mice. A significant decrease in phosphorylated P70S6K levels was also observed in NRG-1(+/-)-Dox compared with nontreated NRG-1(+/-) mice. These results demonstrate that heterozygous knockout of the neuregulin-1 gene worsens survival and LV function in the presence of doxorubicin-induced cardiac injury in vivo. This is associated with the depression of activation of the erbB2 receptor as well as Akt, p70S6K, and ERK-1/2 pathways. PMID- 15833804 TI - Mathematical modeling of the nitric oxide/cGMP pathway in the vascular smooth muscle cell. AB - The nitric oxide (NO)/cGMP pathway in the vascular smooth muscle cell (VSMC) is an important cellular signaling system for the regulation of VSMC relaxation. We present a mathematical model to investigate the underlying mechanisms of this pathway. The model describes the flow of NO-driven signal transduction: NO activation of soluble guanylate cyclase (sGC), sGC- and phosphodiesterase catalyzed cGMP production and degradation, cGMP-mediated regulation of protein targets including the Ca2+-activated K+ (KCa) channel, and the myosin contractile system. Model simulations reproduce major NO/cGMP-induced VSMC relaxation effects, including intracellular Ca2+ concentration reduction and Ca2+ desensitization of myosin phosphorylation and force generation. Using the model, we examine several testable principles. 1) Rapid sGC desensitization is caused by end-product cGMP feedback inhibition; a large fraction of the steady-state sGC population is in an inactivated intermediate state, and cGMP production is limited well below maximum. 2) NO activates the K(Ca) channel with both cGMP dependent and -independent mechanisms; moderate NO concentration affects the K(Ca) via the cGMP-dependent pathway, whereas higher NO concentration is accommodated by a cGMP-independent mechanism. 3) Chronic NO synthase inhibition may cause underexpressions of K+ channels including inward rectifier and K(Ca) channels. 4) Ca2+ desensitization of the contractile system is distinguished from Ca2+ sensitivity of myosin phosphorylation. The model integrates these interactions among the heterogeneous components of the NO signaling system and can serve as a general modeling framework for studying NO-mediated VSMC relaxation under various physiological and pathological conditions. New data can be readily incorporated into this framework for interpretation and possible modification and improvement of the model. PMID- 15833805 TI - Increased myogenic tone in 7-month-old adult male but not female offspring from rat dams exposed to hypoxia during pregnancy. AB - Intrauterine growth restriction (IUGR) increases the risk of cardiovascular disease later in life. Vascular dysfunction occurs in adult offspring from animal models of IUGR including maternal undernutrition, but the influence of reduced fetal oxygen supply on adult vascular function is unclear. Myogenic responses, essential for vascular tone regulation, have not been evaluated in these offspring. We hypothesized that 7-mo-old offspring from hypoxic (12% O(2); H) or nutrient-restricted (40% of control; NR) rat dams would show greater myogenic responses than their 4-mo-old littermates or control (C) offspring through impaired modulation by vasodilators. Growth restriction occurred in male H (P < 0.01), male NR (P < 0.01), and female NR (P < 0.02), but not female H, offspring. Myogenic responses in mesenteric arteries from males but not females were increased at 7 mo in H (P < 0.01) and NR (P < 0.05) vs. C offspring. There was less modulation of myogenic responses after inhibition of nitric oxide synthase (P < 0.05), prostaglandin H synthase (P < 0.005), or both enzymes (P < 0.001) in arteries from 7-mo male H vs. C offspring. Thus reduced vasodilator modulation may explain elevated myogenic responses in 7-mo male H offspring. In contrast, there was increased modulation of myogenic responses in arteries from 7-mo female H vs. C or NR offspring after inhibition of both enzymes (P < 0.05). Thus increased vasodilator modulation may maintain myogenic responses in female H offspring at control levels. In summary, vascular responses in adult offspring from adverse intrauterine environments are impaired in a gender-specific, age dependent, and maternal insult-dependent manner, with males more profoundly affected. PMID- 15833806 TI - Pentoxifylline attenuates cardiac dysfunction and reduces TNF-alpha level in ischemic-reperfused heart. AB - Although pentoxifylline (PTXF), a phosphodiesterase inhibitor, has been reported to exert beneficial effects in cardiac bypass surgery, its effect and mechanisms against ischemia-reperfusion (I/R) injury in heart are poorly understood. Because I/R is known to increase the level of tumor necrosis factor (TNF)-alpha in myocardium and PTXF has been shown to depress the production of TNF-alpha in failing heart, this study examined the hypothesis that PTXF may attenuate cardiac dysfunction and reduce TNF-alpha content in I/R heart. For this purpose, isolated rat hearts were subjected to global ischemia for 30 min followed by reperfusion for 2-30 min. Although cardiac dysfunction due to ischemia was not affected, the recovery of heart function upon reperfusion was markedly improved by PTXF treatment. This cardioprotective effect of PTXF was dose dependent; maximal effect was seen at a concentration of 125 microM. TNF-alpha, nuclear factor kappaB (NF-kappaB), and phosphorylated NF-kappaB contents were decreased in ischemic heart but were markedly increased within 2 min of starting reperfusion. The ratio of cytosolic-to-homogenate NF-kappaB was decreased, whereas the ratio of particulate-to-homogenate NF-kappaB was increased in I/R hearts. These changes in TNF-alpha and NF-kappaB protein contents as well as in NF-kappaB redistribution due to I/R were significantly attenuated by PTXF treatment. The results of this study indicate that the cardioprotective effects of PTXF against I/R injury may be due to reductions in the activation of NF-kappaB and the production of TNF-alpha content. PMID- 15833807 TI - Increased aortic stiffness in the insulin-resistant Zucker fa/fa rat. AB - Accumulating clinical evidence indicates increased aortic stiffness, an independent risk factor for cardiovascular and all-cause mortality, in type 2 diabetic and glucose-intolerant individuals. The present study sought to determine whether increased mechanical stiffness, an altered extracellular matrix, and a profibrotic gene expression profile could be observed in the aorta of the insulin-resistant Zucker fa/fa rat. Mechanical testing of Zucker fa/fa aortas showed increased vascular stiffness in longitudinal and circumferential directions compared with Zucker lean controls. Unequal elevations in developed strain favoring the longitudinal direction resulted in a loss of anisotropy. Real time quantitative PCR and immunohistochemistry revealed increased expression of fibronectin and collagen IV alpha 3 in the Zucker fa/fa aorta. In addition, expression of transforming growth factor-beta and several Smad proteins was increased in vessels from insulin-resistant animals. In rat vascular smooth muscle cells, 12-18 h of exposure to insulin (100 nmol/l) enhanced transforming growth factor-beta1 mRNA expression, implicating a role for hyperinsulinemia in vascular stiffness. Thus there is mechanical, structural, and molecular evidence of arteriosclerosis in the Zucker fa/fa rat at the glucose-intolerant, hyperinsulinemic stage. PMID- 15833808 TI - Angiotensin II AT1 receptors regulate ACE2 and angiotensin-(1-7) expression in the aorta of spontaneously hypertensive rats. AB - When increased in vascular tissues, angiotensin-converting enzyme 2 (ACE2), a carboxypeptidase that hydrolyzes angiotensin II to angiotensin-(1-7), may augment the growth inhibitory and vasodilatory effects of the heptapeptide. We investigated the regulation of ACE2 and angiotensin-(1-7) expression in aortas and carotid arteries of 12-wk-old male spontaneously hypertensive rats (SHR) by determining the effect of sustained angiotensin type 1 (AT(1)) receptor blockade with olmesartan (10 mg.kg(-1).day(-1), n = 13) compared with those that received atenolol (30 mg.kg(-1).day(-1), n = 13), hydralazine (10 mg.kg(-1).day(-1), n = 13), or vehicle (n = 21). Systolic blood pressures were approximately 30% lower (P < 0.05) in rats treated for 2 wk with olmesartan compared with vehicle-treated rats. Both atenolol and hydralazine produced similar decreases in systolic blood pressure. ACE2 mRNA in the thoracic aorta of olmesartan-treated rats (n = 8) was fivefold greater (P < 0.05) than that in vehicle-treated rats (n = 16), whereas atenolol (n = 8) or hydralazine (n = 8) had no effect. Immunostaining intensities in rats treated with olmesartan (n = 5) were also associated with increased (P < 0.05) ACE2 and angiotensin-(1-7) in thoracic aorta media compared with vehicle treated rats. In contrast, immunostaining intensities for both ACE2 and angiotensin-(1-7) were not different from vehicle (n = 5) in carotid arteries of SHR medicated with either atenolol (n = 5) or hydralazine (n = 5). A comparison of vessel wall dimensions showed that olmesartan selectively reduced the thoracic aorta media-to-lumen ratio (P < 0.05) and media thickness (P < 0.05) without an effect on carotid artery morphometry. Compared with vehicle-treated SHR, vascular hypertrophy determined from media and lumen measurements was not changed in SHR given either atenolol or hydralazine. These data represent the first report of ACE2 and angiotensin-(1-7) expression in the aorta and carotid arteries of SHR. Increased ACE2 and angiotensin-(1-7) in association with altered dimensions of the thoracic aorta but not carotid arteries in response to olmesartan treatment provides evidence that this pathway is regulated by AT(1) receptors and may be important in mediating the pressure-independent vascular remodeling effects of angiotensin peptides. PMID- 15833809 TI - Transmural pressure in rat initial subpleural lymphatics during spontaneous or mechanical ventilation. AB - The role played by the mechanical tissue stress in supporting lymph formation and propulsion in thoracic tissues was studied in deeply anesthetized rats (n = 13) during spontaneous breathing or mechanical ventilation. After arterial and venous catheterization and insertion of an intratracheal cannula, fluorescent dextrans were injected intrapleurally to serve as lymphatic markers. After 2 h, the fluorescent intercostal lymphatics were identified, and the hydraulic pressure in lymphatic vessels (P lymph) and adjacent interstitial space (P int) was measured using micropuncture. During spontaneous breathing, end-expiratory P lymph and corresponding P int were -2.5 +/- 1.1 (SE) and 3.1 +/- 0.7 mmHg (P < 0.01), which dropped to -21.1 +/- 1.3 and -12.2 +/- 1.3 mmHg, respectively, at end inspiration. During mechanical ventilation with air at zero end-expiratory alveolar pressure, P lymph and P int were essentially unchanged at end expiration, but, at variance with spontaneous breathing, they increased at end inspiration to 28.1 +/- 7.9 and 28.2 +/- 6.3 mmHg, respectively. The hydraulic transmural pressure gradient (DeltaP tm = P lymph - P int) was in favor of lymph formation throughout the whole respiratory cycle (DeltaP tm = -6.8 +/- 1.2 mmHg) during spontaneous breathing but not during mechanical ventilation (DeltaP tm = 1.1 +/- 1.8 mmHg). Therefore, data suggest that local tissue stress associated with the active contraction of respiratory muscles is required to support an efficient lymphatic drainage from the thoracic tissues. PMID- 15833810 TI - Contribution of Na+-K+ pump and KIR currents to extracellular pH-dependent changes of contractility in rat superior mesenteric artery. AB - We compared the branches and trunk of rat superior mesenteric artery (SMA) with respect to extracellular pH (pHo)-dependent changes in vascular contractility. Decreases in pHo from 7.8 to 6.4 significantly reduced apparent affinity (pD2) to norepinephrine (NE) and maximal contraction by NE, which were more prominent in larger-diameter arteries. On the other hand, decreases in pHo significantly reduced Ba2+-sensitive K+-induced relaxation (which was evoked by elevation of extracellular K+ concentration from 6 to 12 mM) in the first branch and inhibited inwardly rectifying K+ (K(IR)) currents in cultured smooth muscle cells (SMCs) of SMA. RT-PCR revealed transcripts for Kir2.1 in the SMCs. Real-time PCR analysis revealed 6.1-, 3.3-, and 2.2-fold increases in the Kir2.1 mRNA-to-beta-actin mRNA ratios of SMCs of the third, second, and first branches, respectively, vs. the corresponding relative levels of trunk SMCs. The magnitudes of K+-induced relaxation were significantly greater in smaller-diameter arteries, and there was a strong correlation between the transcript levels of Kir2.1 and K+-induced relaxation. A decrease in pHo reduced ouabain-sensitive K+-induced relaxation and ouabain-induced contraction. A decrease in pHo from 7.4 to 6.4 depolarized membrane potential of the cultured SMCs. From these results, we conclude that an increase in pHo activates K(IR) currents and the Na+ -K+ pump, which then reduces vascular contractility. Inasmuch as K(IR) channel densities are significantly greater in smaller-diameter arteries, the reduction in vascular contractility on increasing pHo is more pronounced in smaller-diameter arteries. PMID- 15833811 TI - Time-limited modulation of appetitive Pavlovian memory by D1 and NMDA receptors in the nucleus accumbens. AB - Recent research has implicated the nucleus accumbens (NAc) in consolidating recently acquired goal-directed appetitive memories, including spatial learning and other instrumental processes. However, an important but unresolved issue is whether this forebrain structure also contributes to the consolidation of fundamental forms of appetitive learning acquired by Pavlovian associative processes. In addition, although dopaminergic and glutamatergic influences in the NAc have been implicated in instrumental learning, it is unclear whether similar mechanisms operate during Pavlovian conditioning. To evaluate these issues, the effects of posttraining intra-NAc infusions of D1, D2, and NMDA receptor antagonists, as well as d-amphetamine, were determined on Pavlovian autoshaping in rats, which assesses learning by discriminated approach behavior to a visual conditioned stimulus predictive of food reward. Intracerebral infusions were given either immediately after each conditioning session to disrupt early memory consolidation or after a delay of 24 h. Findings indicate that immediate, but not delayed, infusions of both D1 (SCH 23390) and NMDA (AP-5) receptor antagonists significantly impair learning on this task. By contrast, amphetamine and the D2 receptor antagonist sulpiride were without significant effect. These findings provide the most direct demonstration to date that D1 and NMDA receptors in the NAc contribute to, and are necessary for, the early consolidation of appetitive Pavlovian learning. PMID- 15833812 TI - Corticotropin-releasing hormone activates ERK1/2 MAPK in specific brain areas. AB - Corticotropin-releasing hormone (CRH) coordinates hormonal and behavioral responses to stress. The mitogen-activated protein kinase extracellular signal related kinase 1/2 (ERK1/2) mediates several functions in different forebrain structures and recently has been implicated in CRH signaling in cultured cells. To study in vivo CRH-mediated activation of central ERK1/2, we investigated the expression pattern of the phosphorylated ERK1/2(p-ERK1/2) in the mouse brain after intracerebroventricular CRH injections. As shown by immunohistochemistry and confocal microscopy analysis, CRH administration increased p-ERK1/2 levels specifically in the CA3 and CA1 hippocampal subfields and basolateral complex of the amygdala, both structures related to external environmental information processing and behavioral aspects of stress. Other regions such as hypothalamic nuclei and the central nucleus of the amygdala, also related to central CRH system but involved in the processing of the ascending visceral information and neuroendocrine-autonomic response to stress, did not show CRH-mediated ERK1/2 activation. To dissect the involvement of CRH receptor 1 (CRHR1) and CRHR2, we used conditional knockout mice in which Crhr1 is inactivated in the anterior forebrain and limbic structures. The conditional genetic ablation of Crhr1 inhibited the p-ERK1/2 increase, underlining the involvement of CRHR1 in the CRH mediated activation. These findings underscore the fact that CRH activates p ERK1/2 through CRHR1 only in selected brain regions, pointing to a specific role of this pathway in mediating behavioral adaptation to stress. PMID- 15833813 TI - BCL6b mediates the enhanced magnitude of the secondary response of memory CD8+ T lymphocytes. AB - A characteristic of the secondary response of CD8(+) T cells that distinguishes it from the primary response is the generation of greater numbers of effector cells. Because effector CD8(+) T cells are derived from a pool of less differentiated, replicating cells in secondary lymphoid organs, and because IL-2 mediates effector differentiation, the enhanced secondary response may reflect the enlargement of this generative pool by the transient repression of IL-2 mediated differentiation. We have examined for this function the transcriptional repressor BCL6b, a homologue of BCL6 that represses IL-2-induced B cell differentiation. BCL6b is expressed in a small subset of antigen-experienced CD8(+) T cells. Ectopic expression of BCL6b in CD8(+) T cells diminishes their growth in response to IL-2 in vitro. Female mice in which the BCL6b gene has been interrupted have normal primary responses of CD8(+) T cells to infection with vaccinia expressing the H-Y epitope, Uty, but Uty-specific, BCL6b(-/-), memory CD8(+) T cells have diminished recall proliferative responses to this epitope in vitro. BCL6b(-/-) mice also have normal primary CD8(+) T cell responses to influenza infection, but nucleoprotein peptide-specific, BCL6b(-/-), memory CD8(+) T cells have a cell autonomous defect in the number of effector cells generated in response to reinfection. Therefore, BCL6b is required for the enhanced magnitude of the secondary response of memory CD8(+) T cells. PMID- 15833814 TI - The gene expression signatures of melanoma progression. AB - Because of the paucity of available tissue, little information has previously been available regarding the gene expression profiles of primary melanomas. To understand the molecular basis of melanoma progression, we compared the gene expression profiles of a series of nevi, primary melanomas, and melanoma metastases. We found that metastatic melanomas exhibit two dichotomous patterns of gene expression, which unexpectedly reflect gene expression differences already apparent in comparing laser-capture microdissected radial and vertical phases of a large primary melanoma. Unsupervised hierarchical clustering accurately separated nevi and primary melanomas. Multiclass significance analysis of microarrays comparing normal skin, nevi, primary melanomas, and the two types of metastatic melanoma identified 2,602 transcripts that significantly correlated with sample class. These results suggest that melanoma pathogenesis can be understood as a series of distinct molecular events. The gene expression signatures identified here provide the basis for developing new diagnostics and targeting therapies for patients with malignant melanoma. PMID- 15833815 TI - Adaptation without parameter change: Dynamic gain control in motion detection. AB - Many sensory systems adapt their input-output relationship to changes in the statistics of the ambient stimulus. Such adaptive behavior has been measured in a motion detection sensitive neuron of the fly visual system, H1. The rapid adaptation of the velocity response gain has been interpreted as evidence of optimal matching of the H1 response to the dynamic range of the stimulus, thereby maximizing its information transmission. Here, we show that correlation-type motion detectors, which are commonly thought to underlie fly motion vision, intrinsically possess adaptive properties. Increasing the amplitude of the velocity fluctuations leads to a decrease of the effective gain and the time constant of the velocity response without any change in the parameters of these detectors. The seemingly complex property of this adaptation turns out to be a straightforward consequence of the multidimensionality of the stimulus and the nonlinear nature of the system. PMID- 15833816 TI - Structural basis for antagonism and resistance of bicalutamide in prostate cancer. AB - Carcinoma of the prostate is the most commonly diagnosed cancer in men. The current pharmacological treatment of choice for progressive androgen-dependent prostate cancer is the nonsteroidal antiandrogen, bicalutamide, either as monotherapy or with adjuvant castration or luteinizing hormone-releasing hormone superagonists to block the synthesis of endogenous testosterone. To date, no nonsteroidal or antagonist-bound androgen receptor (AR) structure is available. We solved the x-ray crystal structure of the mutant W741L AR ligand-binding domain bound to R-bicalutamide at 1.8-A resolution. This mutation confers agonist activity to bicalutamide and is likely involved in bicalutamide withdrawal syndrome. The three-dimensional structure demonstrates that the B ring of R bicalutamide in the W741L mutant is accommodated at the location of the indole ring of Trp-741 in the WT AR bound to dihydrotestosterone. Knowledge of the binding mechanism for R-bicalutamide will provide molecular rationale for the development of new antiandrogens and selective AR modulators. PMID- 15833817 TI - Identification of network-level coding units for real-time representation of episodic experiences in the hippocampus. AB - To examine the network-level organizing principles by which the brain achieves its real-time encoding of episodic information, we have developed a 96-channel array to simultaneously record the activity patterns of as many as 260 individual neurons in the mouse hippocampus during various startling episodes. We find that the mnemonic startling episodes triggered firing changes in a set of CA1 neurons in both startle-type and environment-dependent manners. Pattern classification methods reveal that these firing changes form distinct ensemble representations in a low-dimensional encoding subspace. Application of a sliding window technique further enabled us to reliably capture not only the temporal dynamics of real time network encoding but also postevent processing of newly formed ensemble traces. Our analyses revealed that the network-encoding power is derived from a set of functional coding units, termed neural cliques, in the CA1 network. The individual neurons within neural cliques exhibit "collective cospiking" dynamics that allow the neural clique to overcome the response variability of its members and to achieve real-time encoding robustness. Conversion of activation patterns of these coding unit assemblies into a set of real-time digital codes permits concise and universal representation and categorization of discrete behavioral episodes across different individual brains. PMID- 15833818 TI - Deletion of PPARgamma in adipose tissues of mice protects against high fat diet induced obesity and insulin resistance. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a crucial role in adipocyte differentiation, glucose metabolism, and other physiological processes. To further explore the role of PPARgamma in adipose tissues, we used a Cre/loxP strategy to generate adipose-specific PPARgamma knockout mice. These animals exhibited marked abnormalities in the formation and function of both brown and white adipose tissues. When fed a high-fat diet, adipose-specific PPARgamma knockout mice displayed diminished weight gain despite hyperphagia, had diminished serum concentrations of both leptin and adiponectin, and did not develop glucose intolerance or insulin resistance. Characterization of in vivo glucose dynamics pointed to improved hepatic glucose metabolism as the basis for preventing high-fat diet-induced insulin resistance. Our findings further illustrate the essential role for PPARgamma in the development of adipose tissues and suggest that a compensatory induction of hepatic PPARgamma may stimulate an increase in glucose disposal by the liver. PMID- 15833819 TI - Experimental design and analysis of antibody microarrays: applying methods from cDNA arrays. AB - Protein expression microarrays, also called antibody arrays, represent a new technology that allows the expression level of proteins to be assessed directly. As is also the case with gene expression microarrays, it is hoped that protein expression microarrays will aid in biomarker discovery, predicting disease outcomes and response to treatments, and detecting molecular mechanisms and/or pathways associated with a particular disease state. However, accurately achieving these aims is dependent upon suitable experimental designs, normalization procedures that eliminate systematic bias, and appropriate statistical analyses to assess differential expression or expose expression patterns. In the last five years, a large amount of research has been devoted to two-color cDNA arrays to improve experimental design, normalization and statistical analyses to assess differential expression and classification. These methods are directly applicable to two-color antibody arrays. The objective of this article is to discuss statistical methods that have been developed for cDNA arrays and describe how the methods can be directly applied to antibody arrays. PMID- 15833820 TI - Therapeutic targeting of the Hedgehog-GLI pathway in prostate cancer. AB - The Hedgehog-GLI signaling pathway is important in animal development and tumorigenesis. Recent findings indicate that the growth and survival of human prostate cancer cells rely upon sustained signaling from the Hedgehog-GLI pathway. These findings have prompted a novel rational strategy for therapeutic treatment of prostate tumors, including metastatic tumors. PMID- 15833821 TI - Lysosomes as targets for cancer therapy. AB - Tumor invasion and metastasis are associated with altered lysosomal trafficking and increased expression of the lysosomal proteases termed cathepsins. Emerging experimental evidence suggests that such alterations in lysosomes may form an "Achilles heel" for cancer cells by sensitizing them to death pathways involving lysosomal membrane permeabilization and the release of cathepsins into the cytosol. Here, we highlight recent results on cancer-related changes in the composition and function of lysosomes, focusing on possible implications for the development of novel cancer therapeutics that target tumor cell lysosomes. PMID- 15833822 TI - Genetics and Latinos: proceedings of a national summit. PMID- 15833823 TI - The hydra phenomenon of cancer: why tumors recur locally after microscopically complete resection. AB - After surgical resection with microscopically clear margins, solid malignant tumors recur locally in up to 50%. Although the effect of a local tumor recurrence on the overall survival may be low in common cancers such as carcinoma of the breast or prostate, the affected patients suffer from exacerbated fear and the burden of the secondary treatment. With some tumor entities such as carcinoma of the uterine cervix or carcinoma of the head and neck, a local recurrence indicates incurability in the majority of cases. The pathomechanisms of local tumor spread and relapse formation are still unclear and comparatively little research has been devoted to their elucidation. Through the analysis of clinical and molecular data, we propose the concept of two pathogenetically and prognostically different local relapse types (i) in situ recurrences that arise in the residual organ/organ system not involved in the surgery for the primary tumor and (ii) scar recurrences that develop at the site of previous tumor resection. Whereas field cancerization, the monoclonal or multiclonal displacement of normal epithelium by a genetically altered but microscopically undistinguishable homologue, may explain the origin of in situ recurrences, most scar recurrences are regarded as the result of the interaction of minimal residual microscopically occult cancer with the surgical wound environment inside a developmentally defined tissue or organ compartment. The therapeutic implications derived from these concepts and areas of future research aimed to reduce local relapses are discussed in this perspective. PMID- 15833824 TI - Epidermal growth factor receptor dynamics influences response to epidermal growth factor receptor targeted agents. AB - Analysis of gene expression of cancer cell lines exposed to erlotinib, a small molecule inhibitor of the epidermal growth factor receptor (EGFR), showed a marked increase in EGFR mRNA in resistant cell lines but not in susceptible ones. Because cetuximab induces EGFR down-regulation, we explored the hypothesis that treatment with cetuximab would interfere with erlotinib-induced EGFR up regulation and result in antitumor effects. Exposure of the resistant biliary tract cancer cell line HuCCT1 but not the susceptible A431 epidermoid cell line to erlotinib induced EGFR mRNA and protein expression. Combined treatment with cetuximab blunted the erlotinib-induced EGFR up-regulation and resulted in inhibition of cell proliferation and apoptosis in the HuCCT1 cells. Blockage of erlotinib-induced EGFR synthesis in HuCCT1 cells by small interfering RNA resulted in identical antitumor effects as cetuximab, providing mechanistic specificity. In mice xenografted with A431, HuCCT1, and the pancreatic cancer cell line Panc430, maximal growth arrest and decrease in Ki67 proliferation index were documented with combined therapy, and EGFR down-regulation was observed in cetuximab-treated tumors. These results may indicate that resistance to EGFR kinase inhibition may be, at least in part, mediated by a highly dynamic feedback loop consisting of up-regulation of the EGFR upon exposure to EGFR kinase inhibitors. Abrogation of this response by small interfering RNA-mediated EGFR mRNA down-regulation and/or by cetuximab-mediated protein clearance induced tumor arrest across several cancer models with different EGFR expression levels, suggesting that resistance and sensitivity are dynamic events where proportional decrease in the target rather than absolute content dictates outcome. PMID- 15833825 TI - Linking transcriptional elongation and messenger RNA export to metastatic breast cancers. AB - The biochemical pathways that are disrupted in the genesis of sporadic breast cancers remain unclear. Moreover, the present prognosticating markers used to determine the prognosis of node-negative patient leads to probabilistic results, and the eventual clinical course is far from certain. Here we identified the human TREX complex, a multiprotein complex that links transcription elongation to mRNA transport, as culprit of aggressive human breast cancers. We show that whereas p84N5 (called hTREX84) is expressed at very low levels in normal breast epithelial cells, it is highly expressed in breast tumors. Importantly, hTREX84 expression correlates with tumor size and the metastatic state of the tumor progression. Reduction of hTREX84 levels in breast cancer cell lines by small interfering RNA result in inhibition of cellular proliferation and abrogation of mRNA export. These results not only identify hTREX84 as a prognosticator of breast cancer but also delineate human TREX complex as a target for therapeutic drugs against breast cancer. PMID- 15833826 TI - Activation of the Fas-FasL signaling pathway by MDA-7/IL-24 kills human ovarian cancer cells. AB - The tumor-suppressive activity of melanoma differentiation-associated gene-7 (mda 7), also known as interleukin 24 (IL-24), has been shown in a spectrum of human cancer cells in vitro and in vivo. However, mechanisms responsible for antitumor activity of mda-7 in human ovarian cancer cells have not been identified. We investigated the therapeutic activity and underlying mechanisms of adenovirus mediated mda-7 gene (Ad-mda7) transfer in human ovarian cancer cells. Ad-mda7 treatment resulted in overexpression of MDA-7/IL-24 protein in both ovarian cancer and normal ovarian epithelial cells. However, Ad-mda7 significantly (P = 0.001) inhibited cell proliferation and induced apoptosis only in tumor cells and not in normal cells. Studies addressing the mechanism of action of Ad-mda7 induced tumor cell apoptosis revealed early activation of the transcription factors c-Jun and activating transcription factor 2, which in turn stimulated the transcription of an immediate downstream target, the death-inducer Fas ligand (FasL), and its cognate receptor Fas. Associated with the activation of Fas-FasL was the activation of nuclear factor kappaB and induction of Fas-associated factor 1, Fas-associated death domain, and caspase-8. Promoter-based reporter gene analyses showed that Ad-mda7 specifically activated the Fas promoter. Inhibition of Fas using small interfering RNA resulted in a significant decrease in Ad-mda7-mediated tumor cell death. Additionally, blocking of FasL with NOK-1 antibody abrogated Ad-mda7-mediated apoptosis. Collectively, these results show that Ad-mda7-mediated killing of human ovarian cancer cells involves activation of the Fas-FasL signaling pathway, a heretofore unrecognized mediator of MDA-7 apoptosis induction. PMID- 15833827 TI - Stem and progenitor-like cells contribute to the aggressive behavior of human epithelial ovarian cancer. AB - The cellular mechanisms underlying the increasing aggressiveness associated with ovarian cancer progression are poorly understood. Coupled with a lack of identification of specific markers that could aid early diagnoses, the disease becomes a major cause of cancer-related mortality in women. Here we present direct evidence that the aggressiveness of human ovarian cancer may be a result of transformation and dysfunction of stem cells in the ovary. A single tumorigenic clone was isolated among a mixed population of cells derived from the ascites of a patient with advanced ovarian cancer. During the course of the study, yet another clone underwent spontaneous transformation in culture, providing a model of disease progression. Both the transformed clones possess stem cell-like characteristics and differentiate to grow in an anchorage independent manner in vitro as spheroids, although further maturation and tissue specific differentiation was arrested. Significantly, tumors established from these clones in animal models are similar to those in the human disease in their histopathology and cell architecture. Furthermore, the tumorigenic clones, even on serial transplantation continue to establish tumors, thereby confirming their identity as tumor stem cells. These findings suggest that: (a) stem cell transformation can be the underlying cause of ovarian cancer and (b) continuing stochastic events of stem and progenitor cell transformation define the increasing aggression that is characteristically associated with the disease. PMID- 15833828 TI - Metabolic characterization of human prostate cancer with tissue magnetic resonance spectroscopy. AB - Diagnostic advancements for prostate cancer have so greatly increased early detections that hope abounds for improved patient outcomes. However, histopathology, which guides treatment, often subcategorizes aggressiveness insufficiently among moderately differentiated Gleason score (6 and 7) tumors (>70% of new cases). Here, we test the diagnostic capability of prostate metabolite profiles measured with intact tissue magnetic resonance spectroscopy and the sensitivity of local prostate metabolites in predicting prostate cancer status. Prostate tissue samples (n = 199) obtained from 82 prostate cancer patients after prostatectomy were analyzed with high-resolution magic angle spinning proton magnetic resonance spectroscopy, and afterwards with quantitative pathology. Metabolite profiles obtained from principal component analysis of magnetic resonance spectroscopy were correlated with pathologic quantitative findings by using linear regression analysis and evaluated against patient pathologic statuses by using ANOVA. Paired t tests show that tissue metabolite profiles can differentiate malignant from benign samples obtained from the same patient (P < 0.005) and correlate with patient serum prostate-specific antigen levels (P < 0.006). Furthermore, metabolite profiles obtained from histologically benign tissue samples of Gleason score 6 and 7 prostates can delineate a subset of less aggressive tumors (P < 0.008) and predict tumor perineural invasion within the subset (P < 0.03). These results indicate that magnetic resonance spectroscopy metabolite profiles of biopsy tissues may help direct treatment plans by assessing prostate cancer pathologic stage and aggressiveness, which at present can be histopathologically determined only after prostatectomy. PMID- 15833829 TI - Spontaneous human adult stem cell transformation. AB - Human adult stem cells are being evaluated widely for various therapeutic approaches. Several recent clinical trials have reported their safety, showing them to be highly resistant to transformation. The clear similarities between stem cell and cancer stem cell genetic programs are nonetheless the basis of a recent proposal that some cancer stem cells could derive from human adult stem cells. Here we show that although they can be managed safely during the standard ex vivo expansion period (6-8 weeks), human mesenchymal stem cells can undergo spontaneous transformation following long-term in vitro culture (4-5 months). This is the first report of spontaneous transformation of human adult stem cells, supporting the hypothesis of cancer stem cell origin. Our findings indicate the importance of biosafety studies of mesenchymal stem cell biology to efficiently exploit their full clinical therapeutic potential. PMID- 15833830 TI - Ceratamines, structurally simple microtubule-stabilizing antimitotic agents with unusual cellular effects. AB - Ceratamine A and ceratamine B are heterocyclic alkaloids recently identified in a screen for compounds that arrest cells in mitosis. Treatment of breast carcinoma MCF-7 cells causes a concentration-dependent block of cell cycle progression exclusively at mitosis. In vitro studies with purified tubulin indicate that the ceratamines directly stimulate microtubule polymerization in the absence of microtubule-associated proteins. Cells treated with ceratamines show a dense perinuclear microtubule network in interphase and multiple pillar-like tubulin structures in mitotic cells. The ceratamines do not compete with paclitaxel for binding to microtubules in vitro. Unlike other microtubule-stabilizing agents, the ceratamines have simple structures with no chiral centers, making them attractive drug leads. PMID- 15833831 TI - Arginase-producing myeloid suppressor cells in renal cell carcinoma patients: a mechanism of tumor evasion. AB - Myeloid suppressor cells with high arginase activity are found in tumors and spleen of mice with colon and lung cancer. These cells, described as macrophages or immature dendritic cells, deplete arginine and impair T cell proliferation and cytokine production. Although arginase activity has been described in cancer patients, it is thought to originate from tumor cells metabolizing arginine to ornithine needed to sustain rapid cell proliferation. The goal of this study was to determine whether myeloid suppressor cells producing high arginase existed in renal cell carcinoma patients. Peripheral blood mononuclear cells from 123 patients with metastatic renal cell carcinoma, prior to treatment, were found to have a significantly increased arginase activity. These patients had a markedly decreased cytokine production and expressed low levels of T cell receptor CD3zeta chain. Cell separation studies showed that the increased arginase activity was limited to a specific subset of CD11b+, CD14-, CD15+ cells with a polymorphonuclear granulocyte morphology and markers, instead of macrophages or dendritic cells described in mouse models. Furthermore, these patients had low levels of arginine and high levels of ornithine in plasma. Depletion of the CD11b+, CD14- myeloid suppressor cells reestablished T cell proliferation and CD3zeta chain expression. These results showed, for the first time, the existence of suppressor myeloid cells producing arginase in human cancer patients. In addition, it supports the concept that blocking arginase may be an important step in the success of immunotherapy. PMID- 15833832 TI - SV40 enhances the risk of malignant mesothelioma among people exposed to asbestos: a molecular epidemiologic case-control study. AB - We conducted a case-control study on asbestos exposure and presence of SV40 in tumor samples of malignant mesotheliomas (MMs) and bladder urotheliomas (BUs). PCR analysis revealed the presence of SV40 DNA (SV40+) in eight (42.1%) MMs and 6 (33.3%) BUs. The odds ratio for MM Asb- and SV40+ was 0.4 [95% confidence interval (95% CI), 0.03-4.0], for Asb+ and SV40- was 3.6 (95% CI, 0.6-21.0), and for Asb+ and SV40+ was 12.6 (95% CI, 1.2-133.9). Our results suggest that SV40 increases the risk of MM among individuals exposed to asbestos. PMID- 15833833 TI - Loss of heterozygosity in childhood acute lymphoblastic leukemia detected by genome-wide microarray single nucleotide polymorphism analysis. AB - Loss of heterozygosity (LOH) is detectable in many forms of malignancy, including leukemia, using techniques such as microsatellite analysis and comparative genomic hybridization. However, these techniques are laborious and require the use of relatively large amounts of DNA if the whole genome is to be examined. Here we describe the use of oligonucleotide microarrays to characterize single nucleotide polymorphisms (SNPs) in lymphoblasts isolated from children with acute lymphoblastic leukemia for the pan-genomic mapping of LOH with a resolution of 100 to 200 kb. Results were compared with DNA obtained during remission and on relapse. Abnormalities were seen in 8 of 10 cases. The two cases with no abnormalities and one case that showed identical changes at relapse and presentation remain in remission 1 to 9 years following retreatment. The remaining seven patients died following relapse. In four cases, LOH was only detectable at relapse suggesting that progressive LOH may be a cause of disease progression and/or drug resistance. This was supported by detailed analysis of one case in which LOH involving the glucocorticoid receptor was associated with mutation of the remaining allele. The most frequent abnormality detected involved chromosome 9p. In each of the four cases where this was observed LOH included the INK4 locus. In three of the four cases, INK4 loss was only observed at relapse, suggesting that this abnormality may be commonly associated with treatment failure. These observations show that SNP array analysis is a powerful new tool for the analysis of allelic imbalance in leukemic blasts. PMID- 15833834 TI - Serum soluble epidermal growth factor receptor concentrations decrease in postmenopausal metastatic breast cancer patients treated with letrozole. AB - Previous studies have implicated estrogen as a regulator of epidermal growth factor receptor (EGFR) expression in breast tumors. We therefore speculated that estrogen might modulate serologic soluble EGFR (sEGFR) concentrations in breast cancer patients. Accordingly, we measured serum sEGFR concentrations in postmenopausal women with metastatic breast cancer (MBC) treated with letrozole, an aromatase inhibitor that blocks estrogen synthesis. Serum specimens were obtained prior to and following 1 and 3 months of letrozole therapy. We report that sEGFR concentrations do not differ between MBC patients prior to letrozole treatment and age- and postmenopause-matched healthy women (P = 0.468). In contrast, however, sEGFR concentrations decreased significantly in 76% of MBC patients after both 1 month (P = 0.006) and 3 months (P = 0.003) of letrozole therapy versus pretreatment concentrations. Within the limitations of this study, we found no evidence for an association between pretreatment sEGFR concentrations or decreased treatment sEGFR concentrations and either progression-free or overall survival. Nonetheless, we conclude that future prospective studies are warranted to determine if baseline and/or longitudinal serum sEGFR concentrations may be useful for predicting disease progression and survival, and/or for monitoring responsiveness to aromatase inhibitors or other endocrine therapies in breast cancer patients. PMID- 15833835 TI - Gene expression profiling allows distinction between primary and metastatic squamous cell carcinomas in the lung. AB - Lung neoplasms commonly develop in patients previously treated for head and neck carcinomas. The derivation of these tumors, either as new primary lung cancers or as metastatic head and neck cancers, is difficult to establish based on clinical or histopathologic criteria since both are squamous cell carcinomas and have identical features under light microscopy. However, this distinction has significant treatment and prognostic implications. Gene expression profiling was performed on a panel of 52 sequentially collected patients with either primary lung (n = 21) or primary head and neck (n = 31) carcinomas using the Affymetrix HG_U95Av2 high-density oligonucleotide microarray. Unsupervised hierarchical clustering with Ward linkage and the Pearson correlation metric was performed. To assess robustness, bootstrap resampling was performed with 1,000 iterations. A t test of the normalized values for each gene was used to determine the genes responsible for segregating head and neck from lung primary carcinomas, and those with the most differential expression were used for later analyses. In the absence of a large "test" set of tumors, we used a supervised leave-one-out cross validation to test how well we could predict the tumor origin. Once a gene expression profile was established, 12 lung lesions taken from patients with previously treated head and neck cancers were similarly analyzed by gene expression profiling to determine their sites of origin. Unsupervised clustering analysis separated the study cohort into two distinct groups which reliably remained segregated with bootstrap resampling. Group 1 consisted of 30 tongue carcinomas. Group 2 consisted of 21 lung cancers and 1 tongue carcinoma. The clustering was not changed even when normal lung or tongue profiles were subtracted from the corresponding carcinomatous lesions, and a leave-one-out cross-validation showed a 98% correct prediction (see Supplementary Data 1). A minimum set of 500 genes required to distinguish these groups was established. Given the ability to segregate these lesions using molecular profiling, we analyzed the lung tumors of undetermined origin. All cases clearly clustered with either lung or tongue tumor subsets, strongly supporting our hypothesis that this technique could elucidate the tissue of origin of metastatic lesions. Although histologically similar, squamous cell carcinomas have distinct gene expression profiles based on their anatomic sites of origin. Accordingly, the application of gene expression profiling may be useful in identifying the derivation of lung nodules and consequently enhances treatment planning. PMID- 15833836 TI - Expression of the Brn-3b transcription factor correlates with expression of HSP 27 in breast cancer biopsies and is required for maximal activation of the HSP-27 promoter. AB - In breast cancer, overexpression of the small heat shock protein, HSP-27, is associated with increased anchorage-independent growth, increased invasiveness, and resistance to chemotherapeutic drugs and is associated with poor prognosis and reduced disease-free survival. Therefore, factors that increase the expression of HSP-27 in breast cancer are likely to affect the prognosis and outcome of treatment. In this study, we show a strong correlation between elevated levels of the Brn-3b POU transcription factor and high levels of HSP-27 protein in manipulated MCF-7 breast cancer cells as well as in human breast biopsies. Conversely, HSP-27 is decreased on loss of Brn-3b. In cotransfection assays, Brn-3b can strongly transactivate the HSP-27 promoter, supporting a role for direct regulation of HSP-27 expression. Brn-3b also cooperates with the estrogen receptor (ER) to facilitate maximal stimulation of the HSP-27 promoter, with significantly enhanced activity of this promoter observed on coexpression of Brn-3b and ER compared with either alone. RNA interference and site-directed mutagenesis support the requirement for the Brn-3b binding site on the HSP-27 promoter, which facilitates maximal transactivation either alone or on interaction with the ER. Chromatin immunoprecipitation provides evidence for association of Brn-3b with the HSP-27 promoter in the intact cell. Thus, Brn-3b can, directly and indirectly (via interaction with the ER), activate HSP-27 expression, and this may represent one mechanism by which Brn-3b mediates its effects in breast cancer cells. PMID- 15833837 TI - Evidence for the presence of disease-perturbed networks in prostate cancer cells by genomic and proteomic analyses: a systems approach to disease. AB - Prostate cancer is initially responsive to androgen ablation therapy and progresses to androgen-unresponsive states that are refractory to treatment. The mechanism of this transition is unknown. A systems approach to disease begins with the quantitative delineation of the informational elements (mRNAs and proteins) in various disease states. We employed two recently developed high throughput technologies, massively parallel signature sequencing (MPSS) and isotope-coded affinity tag, to gain a comprehensive picture of the changes in mRNA levels and more restricted analysis of protein levels, respectively, during the transition from androgen-dependent LNCaP (model for early-stage prostate cancer) to androgen-independent CL1 cells (model for late-stage prostate cancer). We sequenced >5 million MPSS signatures, obtained >142,000 tandem mass spectra, and built comprehensive MPSS and proteomic databases. The integrated mRNA and protein expression data revealed underlying functional differences between androgen-dependent and androgen-independent prostate cancer cells. The high sensitivity of MPSS enabled us to identify virtually all of the expressed transcripts and to quantify the changes in gene expression between these two cell states, including functionally important low-abundance mRNAs, such as those encoding transcription factors and signal transduction molecules. These data enable us to map the differences onto extant physiologic networks, creating perturbation networks that reflect prostate cancer progression. We found 37 BioCarta and 14 Kyoto Encyclopedia of Genes and Genomes pathways that are up regulated and 23 BioCarta and 22 Kyoto Encyclopedia of Genes and Genomes pathways that are down-regulated in LNCaP cells versus CL1 cells. Our efforts represent a significant step toward a systems approach to understanding prostate cancer progression. PMID- 15833838 TI - Overexpressed P-cadherin/CDH3 promotes motility of pancreatic cancer cells by interacting with p120ctn and activating rho-family GTPases. AB - P-Cadherin/CDH3 belongs to the family of classic cadherins that are engaged in various cellular activities including motility, invasion, and signaling of tumor cells, in addition to cell adhesion. However, the biological roles of P-cadherin itself are not fully characterized. Based on information derived from a previous genome-wide cDNA microarray analysis of microdissected pancreatic ductal adenocarcinoma (PDAC), we focused on P-cadherin as one of the genes most strongly overexpressed in the great majority of PDACs. To investigate the consequences of overexpression of P-cadherin in terms of pancreatic carcinogenesis and tumor progression, we used a P-cadherin-deficient PDAC cell line, Panc-1, to construct a cell line (Panc1-CDH3) that stably overexpressed P-cadherin. Induction of P cadherin in Panc1-CDH3 increased the motility of the cancer cells, but a blocking antibody against P-cadherin suppressed the motility in vitro. Overexpression of P cadherin was strongly associated with cytoplasmic accumulation of one of the catenins, p120ctn, and cadherin switching in PDAC cells. Moreover, P-cadherin dependent activation of cell motility was associated with activation of Rho GTPases, Rac1 and Cdc42, through accumulation of p120ctn in cytoplasm and cadherin switching. These findings suggest that overexpression of P-cadherin is likely to be related to the biological aggressiveness of PDACs; blocking of P cadherin activity or its associated signaling could be a novel therapeutic approach for treatment of aggressive pancreatic cancers. PMID- 15833839 TI - Influence of mast cells on structural and functional manifestations of radiation induced heart disease. AB - Radiation-induced heart disease (RIHD), characterized by accelerated atherosclerosis and adverse tissue remodeling, is a serious sequelae after radiotherapy of thoracic and chest wall tumors. Adverse cardiac remodeling in RIHD and other cardiac disorders is frequently accompanied by mast cell hyperplasia, suggesting that mast cells may affect the development of cardiac fibrosis. This study used a mast cell-deficient rat model to define the role of mast cells in RIHD. Mast cell-deficient rats (Ws/Ws) and mast cell-competent littermate controls (+/+) were exposed to 18 Gy localized single-dose irradiation of the heart. Six months after irradiation, cardiac function was examined by echocardiography and Langendorff-perfused isolated heart preparation, whereas structural changes were assessed using quantitative histology and immunohistochemical analysis. Mast cell-deficient rats exhibited more severe postradiation changes than mast cell-competent littermates. Hence, mast cell deficient rats exhibited a greater upward/leftward shift in the left ventricular (LV) diastolic pressure-volume relationship (P = 0.001), a greater reduction in in vivo LV diastolic area (from 0.50 +/- 0.024 cm in age-matched controls to 0.24 +/- 0.032 cm after irradiation; P = 0.006), and a greater increase in LV posterior wall thickness (from 0.13 +/- 0.003 cm in age-matched controls to 0.15 +/- 0.003 cm after irradiation; P = 0.04). Structural analysis revealed more pronounced postradiation accumulation of interstitial collagen III but less myocardial degeneration in hearts from mast cell-deficient rats. These data show that the absence of mast cells accelerates the development of functional changes in the irradiated heart, particularly diastolic dysfunction, and suggest that, in contrast to what has been the prevailing assumption, the role of mast cells in RIHD is predominantly protective. PMID- 15833840 TI - The serine/threonine protein kinase, p90 ribosomal S6 kinase, is an important regulator of prostate cancer cell proliferation. AB - An increase in the activity of mitogen-activated protein kinase (MAPK) has been correlated with the progression of prostate cancer to advanced disease in humans. The serine/threonine protein kinase p90-kDa ribosomal S6 kinase (RSK) is an important downstream effector of MAPK but its role in prostate cancer has not previously been examined. Increasing RSK isoform 2 (RSK2) levels in the human prostate cancer line, LNCaP, enhanced prostate-specific antigen (PSA) expression, an important diagnostic marker for prostate cancer, whereas inhibiting RSK activity using a RSK-specific inhibitor, 3Ac-SL0101, decreased PSA expression. The RSK2 regulation of PSA expression occurred via a mechanism involving both RSK2 kinase activity and its ability to associate with the coactivator, p300. RNA interference of the androgen receptor (AR) showed that the AR was important in the RSK2-mediated increase in PSA expression. RSK levels are higher in approximately 50% of human prostate cancers compared with normal prostate tissue, which suggests that increased RSK levels may participate in the rise in PSA expression that occurs in prostate cancer. Furthermore, 3Ac-SL0101 inhibited proliferation of the LNCaP line and the androgen-independent human prostate cancer line, PC-3. These results suggest that proliferation of some prostate cancer cells is dependent on RSK activity and support the hypothesis that RSK may be an important chemotherapeutic target for prostate cancer. PMID- 15833841 TI - RRP22 is a farnesylated, nucleolar, Ras-related protein with tumor suppressor potential. AB - Ras proteins are members of a superfamily of related small GTPases. Some members, such as Ras, are oncogenic. However, other members seem to serve as tumor suppressors, such as Rig and Noey2. We now identify and characterize a novel member of the Ras superfamily, RRP22. Like Ras, RRP22 can be posttranslationally modified by farnesyl. Unlike Ras, RRP22 inhibits cell growth and promotes caspase independent cell death. Examination of human tumor cells shows that RRP22 is frequently down-regulated due to promoter methylation. Moreover, reexpression of RRP22 in an RRP22-negative neural tumor cell line impairs its growth in soft agar. Unusually for a Ras-related protein, RRP22 localizes to the nucleolus in a GTP-dependent manner, suggesting a novel mechanism of action. Thus, we identify a new member of the Ras superfamily that can serve as a potential tumor suppressor. PMID- 15833842 TI - Tumorigenic heterogeneity in cancer stem cells evolved from long-term cultures of telomerase-immortalized human mesenchymal stem cells. AB - Long-term cultures of telomerase-transduced adult human mesenchymal stem cells (hMSC) may evolve spontaneous genetic changes leading to tumorigenicity in immunodeficient mice (e.g., hMSC-TERT20). We wished to clarify whether this unusual phenotype reflected a rare but dominant subpopulation or if the stem cell origin allowed most cells to behave as cancer stem cells. Cultures of the hMSC TERT20 strain at population doubling 440 were highly clonogenic (94%). From 110 single-cell clones expanded by 20 population doublings, 6 underwent detailed comparison. Like the parental population, each clone had approximately 1.2 days doubling time with loss of contact inhibition. All retained 1,25-(OH)(2) vitamin D(3)-induced expression of osteoblastic markers: collagen type I, alkaline phosphatase, and osteocalcin. All shared INK4a/ARF gene locus deletion and epigenetic silencing of the DBCCR1 tumor suppressor gene. Despite in vitro commonality, only four of six clones shared the growth kinetics and 100% tumorigenicity of the parental population. In contrast, one clone consistently formed latent tumors and the other established tumors with only 30% penetrance. Changing the in vitro microenvironment to mimic in vivo growth aspects revealed concordant clonal heterogeneity. Latent tumor growth correlated with extracellular matrix entrapment of multicellular spheroids and high procollagen type III expression. Poor tumorigenicity correlated with in vitro serum dependence and high p27(Kip1) expression. Aggressive tumorigenicity correlated with good viability plus capillary morphogenesis on serum starvation and high cyclin D1 expression. Thus, hMSC-TERT20 clones represent cancer stem cells with hierarchical tumorigenicity, providing new models to explore the stem cell hypothesis for cancer. PMID- 15833843 TI - Inhibition of a new differentiation pathway in neuroblastoma by copy number defects of N-myc, Cdc42, and nm23 genes. AB - The best studied oncogenic mechanisms are inactivating defects in both alleles of tumor suppressor genes and activating mutations in oncogenes. Chromosomal gains and losses are frequent in human tumors, but for many regions, like 1p36 and 17q in neuroblastoma, no mutated tumor suppressor genes or oncogenes were identified. Amplification of N-myc in neuroblastoma is strongly correlated with loss of 1p36 and gain of 17q. Here we report that N-myc down-regulates the mRNA expression of many genes with a role in cell architecture. One of them is the 1p36 gene Cdc42. Restoring the Cdc42 expression in neuroblastoma cells strongly induced differentiation. N-myc also inhibited Cdc42 functioning at the protein level. This was mediated by nm23-H1 and nm23-H2, which are located in the amplified 17q region. Nm23-H1 and nm23-H2 are strongly up-regulated downstream targets of N myc. Nm23-H1 was shown to bind Cdc42 and prevented the induction of differentiation. Overexpression of Nm23 due to gain of 17q and induction by N-myc combined with weak expression of Cdc42 due to loss of 1p36 and down-regulation by N-myc can thus block differentiation. Although this marks Cdc42 as a candidate tumor suppressor gene, no mutations were found. Further silencing of Cdc42 by small interfering RNA induced massive apoptosis, indicating that tumor cell survival requires a minimal Cdc42 activity. Three regions of chromosomal gain and loss thus affect genes functioning in one pathway in neuroblastoma. They converge to bring the pathway out of balance and prevent Cdc42 mediated differentiation. PMID- 15833844 TI - Progression of Barrett's metaplasia to adenocarcinoma is associated with the suppression of the transcriptional programs of epidermal differentiation. AB - We did expressional profiling on 24 paired samples of normal esophageal epithelium, Barrett's metaplasia, and esophageal adenocarcinomas. Matching tissue samples representing the three different histologic types were obtained from each patient undergoing esophagectomy for adenocarcinoma. Our analysis compared the molecular changes accompanying the transformation of normal squamous epithelium with Barrett's esophagus and adenocarcinoma in individual patients rather than in a random cohort. We tested the hypothesis that expressional profiling may reveal gene sets that can be used as molecular markers of progression from normal esophageal epithelium to Barrett's esophagus and adenocarcinoma. Expressional profiling was done using U133A GeneChip (Affymetrix), which represent approximately two thirds of the human genome. The final selection of 214 genes permitted the discrimination of differential gene expression of normal esophageal squamous epithelium, Barrett's esophagus, and adenocarcinoma using two dimensional hierarchical clustering of selected genes. These data indicate that transformation of Barrett's esophagus to adenocarcinoma is associated with suppression of the genes involved in epidermal differentiation, including genes in 1q21 loci and corresponding to the epidermal differentiation complex. Correlation analysis of genes concordantly expressed in Barrett's esophagus and adenocarcinoma revealed 21 genes that represent potential genetic markers of disease progression and pharmacologic targets for treatment intervention. PCR analysis of genes selected based on DNA array experiments revealed that estimation of the ratios of GATA6 to SPRR3 allows discrimination among normal esophageal epithelium, Barrett's dysplasia, and adenocarcinoma. PMID- 15833845 TI - Immortal DNA strand cosegregation requires p53/IMPDH-dependent asymmetric self renewal associated with adult stem cells. AB - Because they are long-lived and cycle continuously, adult stem cells (ASCs) are predicted as the most common precursor for cancers in adult mammalian tissues. Two unique attributes have been proposed to restrict the carcinogenic potential of ASCs. These are asymmetric self-renewal that limits their number and immortal DNA strand cosegregation that limits their accumulation of mutations due to DNA replication errors. Until recently, the molecular basis and regulation of these important ASC-specific functions were unknown. We developed engineered cultured cells that exhibit asymmetric self-renewal and immortal DNA strand cosegregation. These model cells were used to show that both ASC-specific functions are regulated by the p53 cancer gene. Previously, we proposed that IMP dehydrogenase (IMPDH) was an essential factor for p53-dependent asymmetric self-renewal. We now confirm this proposal and provide quantitative evidence that asymmetric self renewal is acutely sensitive to even modest changes in IMPDH expression. These analyses reveal that immortal DNA strand cosegregation is also regulated by IMPDH and confirm the original implicit precept that immortal DNA strand cosegregation is specific to cells undergoing asymmetric self-renewal (i.e., ASCs). With IMPDH being the rate-determining enzyme for guanine ribonucleotide (rGNP) biosynthesis, its requirement implicates rGNPs as important regulators of ASC asymmetric self renewal and immortal DNA strand cosegregation. An in silico analysis of global gene expression data from human cancer cell lines underscored the importance of p53-IMPDH-rGNP regulation for normal tissue cell kinetics, providing further support for the concept that ASCs are key targets for adult tissue carcinogenesis. PMID- 15833846 TI - Down-regulation of mitochondrial F1F0-ATP synthase in human colon cancer cells with induced 5-fluorouracil resistance. AB - 5-Fluorouracil (5-FU) is widely used for treatment of advanced colorectal cancer. However, it is common for such patients to develop resistance to 5-FU, and this drug resistance becomes a critical problem for chemotherapy. The mechanisms underlying this resistance are largely unknown. To screen for proteins possibly responsible for 5-FU resistance, cells resistant to 5-FU were derived from human colon cancer cell lines and two-dimensional gel electrophoresis-based comparative proteomics was done. Two-dimensional gel electrophoresis data showed there was lower expression of the alpha subunit of mitochondrial F(1)F(0)-ATP synthase (ATP synthase) in 5-FU-resistant cells compared with parent cells. Western blotting showed that expression of other ATP synthase complex subunits was also lower in 5 FU-resistant cell lines and that these resistant cells also showed decreased ATP synthase activity and reduced intracellular ATP content. The ATP synthase inhibitor, oligomycin A, strongly antagonized 5-FU-induced suppression of cell proliferation. When 5-FU sensitivity was compared with ATP synthase activity in six different human colon cancer cell lines, a positive correlation has been found. Furthermore, suppressed ATP synthase d-subunit expression by siRNA transfection increased cell viability in the presence of 5-FU. Bioenergetic dysfunction of mitochondria has been reported as a hallmark of many types of cancers (i.e., down-regulation of ATP synthase beta-subunit expression in liver, kidney, colon, squamous oesophageal, and lung carcinomas, as well as in breast and gastric adenocarcinomas). Our findings show that ATP synthase down-regulation may not only be a bioenergetic signature of colorectal carcinomas but may also lead to cellular events responsible for 5-FU resistance. PMID- 15833847 TI - Anoxia is necessary for tumor cell toxicity caused by a low-oxygen environment. AB - Cells exposed to oxygen deprivation in vitro have been shown to reduce proliferation and/or engage in programmed cell death. There is considerable controversy in the literature as to the role of hypoxia-inducible factor-1 (HIF 1) and HIF-1 target genes in initiating these responses. We therefore examined the oxygen dependence and the role of the hypoxia-responsive transcription factor HIF-1 in making the cellular death decision. Oxygen concentrations as low as 0.5% did not alter the growth of HIF-1-proficient or HIF-1-deficient murine fibroblasts, or human tumor cells, despite the appropriate induction of HIF-1 target genes. Severe hypoxia (<0.01% oxygen) did induced apoptosis, resulting in decreased colony formation, chromatin condensation, DNA fragmentation, and caspase activation but also independent of HIF1alpha status. Transcriptional induction of HIF-1-dependent genes putatively involved in cell death like BNip3 and BNip3L was therefore disassociated from hypoxia-dependent toxicity. Likewise, forced overexpression of a nondegradable form of HIF-1alpha in several human tumor cell lines was not sufficient to induce apoptosis under normoxic conditions. Taken together, these findings indicate that additional molecular events are triggered by anoxia in a HIF-1-independent manner, and these changes are necessary for cell death observed in low-oxygen environments. PMID- 15833848 TI - Pak1 phosphorylation of snail, a master regulator of epithelial-to-mesenchyme transition, modulates snail's subcellular localization and functions. AB - The process of epithelial-mesenchymal transition plays a pivotal role in the conversion of early stage tumors into invasive malignancies, and has been shown to be regulated by the zinc finger phosphoprotein, Snail; however, no upstream signaling kinases have been shown to modulate Snail functions. Since the invasiveness of breast cancer cells is also influenced by p21-activated kinase 1 (Pak1) signaling, we investigated Pak1's potential mechanistic role in the regulation of Snail functions. We found for the first time that Pak1 promotes transcription repression activity of Snail from E-cadherin, occludin, and aromatase promoters. Pak1 regulates the repressor activity of Snail by phosphorylating on Ser(246). Pak1 phosphorylation of Snail supports Snail's accumulation in the nucleus as well as its repressor functions. A Ser(246)Ala substitution in Snail or Pak1 knockdown by short interference RNA blocked Pak1 mediated Snail phosphorylation, leading to increased cytoplasmic accumulation of Snail and attenuation of Snail repressor activity in breast cancer cells. The regulation of phosphorylation and function of Snail by Pak1 represents a novel mechanism by which a signaling kinase might contribute to the process of epithelial-mesenchymal transition. PMID- 15833849 TI - Fetal stromal-dependent paracrine and intracrine vascular endothelial growth factor-a/vascular endothelial growth factor receptor-1 signaling promotes proliferation and motility of human primary myeloma cells. AB - Induction of neoangiogenesis plays an important role in the pathogenesis of multiple myeloma. However, the mechanism by which expression of vascular endothelial growth factor (VEGF)-A and its receptors modulate the interaction of multiple myeloma cells with stromal cells is not known. Here, we describe a novel in vitro coculture system using fetal bone stromal cells as a feeder layer, which facilitates the survival and growth of human primary multiple myeloma cells. We show that stromal-dependent paracrine VEGF-A signaling promotes proliferation of human primary multiple myeloma cells. Primary multiple myeloma cells only expressed functional VEGF receptor (VEGFR)-1, but not VEGFR-2 or VEGFR-3. VEGFR-1 expression was detected in the cytoplasm and the nuclei of proliferating multiple myeloma cells. Inhibition of VEGFR-1 abrogated multiple myeloma cell proliferation and motility, suggesting that the functional interaction of VEGF-A with its cognate receptor is essential for the growth of primary multiple myeloma cells. Collectively, our results suggest that stromal-dependent paracrine and intracrine VEGF-A/VEGFR-1 signaling contributes to human primary multiple myeloma cell growth and therefore, VEGFR-1 blockade is a potential therapeutic strategy for the treatment of multiple myeloma. PMID- 15833850 TI - Extracellular matrix metalloproteinase inducer stimulates tumor angiogenesis by elevating vascular endothelial cell growth factor and matrix metalloproteinases. AB - Matrix metalloproteinases (MMPs) are endopeptidases that play pivotal roles in promoting tumor disease progression, including tumor angiogenesis. In many solid tumors, MMP expression could be attributed to tumor stromal cells and is partially regulated by tumor-stroma interactions via tumor cell-associated extracellular matrix metalloproteinase inducer (EMMPRIN). The role of EMMPRIN during tumor angiogenesis and growth was explored by modulating EMMPRIN expression and activity using recombinant DNA engineering and neutralizing antibodies. In human breast cancer cells, changes in EMMPRIN expression influenced vascular endothelial growth factor (VEGF) production at both RNA and protein levels. In coculture of tumor cells and fibroblasts mimicking tumor stroma interactions, VEGF expression was induced in an EMMPRIN- and MMP-dependent fashion, and was further enhanced by overexpressing EMMPRIN. Conversely, VEGF expression was inhibited by suppressing EMMPRIN expression in tumor cells, by neutralizing EMMPRIN activity, or by inhibiting MMPs. In vivo, EMMPRIN overexpression stimulated tumor angiogenesis and growth; both were significantly inhibited by antisense suppression of EMMPRIN. Expression of both human and mouse VEGF and MMP, derived from tumor and host cells, respectively, was regulated by EMMPRIN. These results suggest a novel tumor angiogenesis mechanism in which tumor-associated EMMPRIN functionally mediates tumor-stroma interactions and directly contributes to tumor angiogenesis and growth by stimulating VEGF and MMP expression. PMID- 15833851 TI - The contribution of bone marrow-derived cells to the tumor vasculature in neuroblastoma is matrix metalloproteinase-9 dependent. AB - The contribution of the tumor stroma to cancer progression has been increasingly recognized. We had previously shown that in human neuroblastoma tumors orthotopically implanted in immunodeficient mice, stromal-derived matrix metalloproteinase-9 (MMP-9) contributes to the formation of a mature vasculature by promoting pericyte recruitment along endothelial cells. Here we show that MMP 9 is predominantly expressed by bone marrow-derived CD45-positive leukocytes. Using a series of bone marrow transplantation experiments in MMP-9(+/+) and MMP 9(-/-) mice xenotransplanted with human neuroblastoma tumors, we show that bone marrow-derived MMP-9 is critical for the recruitment of leukocytes from bone marrow into the tumor stroma and for the integration of bone marrow-derived endothelial cells into the tumor vasculature. Expression of MMP-9 by bone marrow derived cells in the tumor stroma is also critical for the formation of a mature vasculature and coverage of endothelial cells with pericytes. Furthermore, in primary human neuroblastoma tumor specimens of unfavorable histology, we observed a higher level of tumor infiltration with MMP-9 expressing phagocytic cells and a higher degree of coverage of endothelial cells by pericytes when compared with tumor specimens with a favorable histology. Taken together, the data show that in neuroblastoma, MMP-9 plays a critical role in the recruitment of bone marrow derived cells to the tumor microenvironment where they positively contribute to angiogenesis and tumor progression. PMID- 15833852 TI - Nuclear factor-kappaB-dependent mechanisms in breast cancer cells regulate tumor burden and osteolysis in bone. AB - A central mediator of a wide host of target genes, the nuclear factor-kappaB (NF kappaB) family of transcription factors, has emerged as a molecular target in cancer and diseases associated with bone destruction. To evaluate how NF-kappaB signaling in tumor cells regulates processes associated with osteolytic bone tumor burden, we stably infected the bone-seeking MDA-MB-231 breast cancer cell line with a dominant-negative mutant IkappaB that prevents phosphorylation of IkappaBalpha and associated nuclear translocation of NF-kappaB. Blockade of NF kappaB signaling in MDA-MB-231 cells by the mutant IkappaB decreased in vitro cell proliferation, expression of the proinflammatory, bone-resorbing cytokine interleukin-6, and in vitro bone resorption by tumor/osteoclast cocultures while reciprocally up-regulating production of the proapoptotic enzyme caspase-3. Suppression of NF-kappaB transcription in these breast cancer cells also reduced incidence of in vivo tumor-mediated osteolysis after intratibial injection of tumor cells in female athymic nude mice. Immunohistochemistry showed that the cancerous lesions formed in bone by MDA-MB-231 cells express both interleukin-6 and the p65 subunit of NF-kappaB at the bone-tumor interface. NF-kappaB signaling in breast cancer cells therefore promotes bone tumor burden and tumor-mediated osteolysis through combined control of tumor proliferation, cell survival, and bone resorption. These findings imply that NF-kappaB and its associated genes may be relevant therapeutic targets in osteolytic tumor burden. PMID- 15833853 TI - High nuclear S100A6 (Calcyclin) is significantly associated with poor survival in pancreatic cancer patients. AB - Recent studies have reported elevated levels of S100A6 in pancreatic ductal adenocarcinoma cells. Here, we describe a detailed analysis of S100A6 expression in benign (n = 32), malignant (n = 60), and premalignant pancreatic ductal cells [96 pancreatic intraepithelial neoplasias (PanIN) from 46 patients]. S100A6 staining was more intense in malignant cells than in benign cells (P = 0.0001). In malignant cells, staining was higher in the nucleus than in the cytoplasm (P = 0.003). Univariate analysis revealed a significant decrease in survival time for patients with high levels of nuclear (P = 0.01) but not cytoplasmic (P = 0.20) S100A6. No evidence was found for an association between nuclear S100A6 expression and other variables, including gender, age at surgery, tumor size or grade, nodal metastases, resection margin, vascular invasion, perineural invasion, p53 or Smad4 levels (both linked to survival in previous studies), or the p65 subunit of nuclear factor-kappaB (a potential regulator of S100A6). Although nodal metastases and resection margin involvement were also associated with poor survival (P = 0.06 in both cases), multivariate analysis suggests that nuclear S100A6 is a significant independent indicator of survival (P = 0.003). Whereas PanIN 1a lesions showed a general absence of S100A6 staining, there was a progressive increase in the proportion of positively stained PanINs with increasing PanIN grade. In particular, we observed an increase in the frequency and intensity of nuclear staining. Our results suggest that up-regulation of S100A6 is an early event in pancreatic cancer development and that elevated levels of nuclear S100A6 may affect clinical outcome. PMID- 15833854 TI - Inhibition of mammalian target of rapamycin reverses alveolar epithelial neoplasia induced by oncogenic K-ras. AB - The serine/threonine kinase AKT and its downstream mediator mammalian target of rapamycin (mTOR) are activated in lung adenocarcinoma, and clinical trials are under way to test whether inhibition of mTOR is useful in treating lung cancer. Here, we report that mTOR inhibition blocked malignant progression in K-ras(LA1) mice, which undergo somatic activation of the K-ras oncogene and display morphologic changes in alveolar epithelial cells that recapitulate those of precursors of human lung adenocarcinoma. Levels of phospho-S6(Ser236/235), a downstream mediator of mTOR, increased with malignant progression (normal alveolar epithelial cells to adenocarcinoma) in K-ras(LA1) mice and in patients with lung adenocarcinoma. Atypical alveolar hyperplasia, an early neoplastic change, was prominently associated with macrophages and expressed high levels of phospho-S6(Ser236/235). mTOR inhibition in K-ras(LA1) mice by treatment with the rapamycin analogue CCI-779 reduced the size and number of early epithelial neoplastic lesions (atypical alveolar hyperplasia and adenomas) and induced apoptosis of intraepithelial macrophages. LKR-13, a lung adenocarcinoma cell line derived from K-ras(LA1) mice, was resistant to treatment with CCI-779 in vitro. However, LKR-13 cells grown as syngeneic tumors recruited macrophages, and those tumors regressed in response to treatment with CCI-779. Lastly, conditioned medium from primary cultures of alveolar macrophages stimulated the proliferation of LKR-13 cells. These findings provide evidence that the expansion of lung adenocarcinoma precursors induced by oncogenic K-ras requires mTOR-dependent signaling and that host factors derived from macrophages play a critical role in adenocarcinoma progression. PMID- 15833855 TI - Mitochondrial damage mediates genotoxicity of arsenic in mammalian cells. AB - Arsenic is an important environmental carcinogen that affects millions of people worldwide through contaminated water supplies. For decades, arsenic was considered a nongenotoxic carcinogen. Using the highly sensitive A(L) mutation assay, we previously showed that arsenic is, indeed, a potent gene and chromosomal mutagen and that its effects are mediated through the induction of reactive oxygen species. However, the origin of these radicals and the pathways involved are not known. Here we show that mitochondrial damage plays a crucial role in arsenic mutagenicity. Treatment of enucleated cells with arsenic followed by rescue fusion with karyoplasts from controls resulted in significant mutant induction. In contrast, treatment of mitochondrial DNA-depleted (rho(0)) cells produced few or no mutations. Mitochondrial damage can lead to the release of superoxide anions, which then react with nitric oxide to produce the highly reactive peroxynitrites. The mutagenic damage was dampened by the nitric oxide synthase inhibitor, N(G)-methyl-L-arginine. These data illustrate that mitochondria are a primary target in arsenic-induced genotoxic response and that a better understanding of the mutagenic/carcinogenic mechanism of arsenic should provide a basis for better interventional approach in both treatment and prevention of arsenic-induced cancer. PMID- 15833856 TI - Suppression of ovarian cancer cell tumorigenicity and evasion of Cisplatin resistance using a truncated epidermal growth factor receptor in a rat model. AB - The overexpression of the epidermal growth factor receptor (EGFR) is associated with a poor prognosis in ovarian cancer. The dominant-negative EGFR (EGFR-DNR) is a truncated receptor that lacks the tyrosine kinase domain and is devoid of signaling capability. This study tested the effects of a EGFR-DNR approach in ovarian cancer cells. NuTu-19, a rat ovarian cancer cell line was rendered resistant to cisplatin. Both NuTu-19 and resistant cells were infected with a retroviral vector containing the EGFR-DNR. NuTu-19 and NuTu-DNR (NuTu-19 cells expressing the EGFR-DNR) were injected into Fisher 344 immunocompetent rats. Western blot analyses were used to assess signal transduction pathways. All rats injected with NuTu-DNR cells remained healthy following tumor injection. In contrast, 100% of the rats injected with the NuTu-19 and NuTu-Sham (NuTu-19 cells expressing an empty vector) died of disease progression at the end of 15 weeks (P = 0.00009). On Western blot analysis, both NuTu-19 and NuTu-Sham cells showed a strong activation of mitogen-activated protein kinase (MAPK) after exposure to EGF. Cisplatin-resistant cell lines showed an enhanced EGF stimulatory effect via the MAPK pathway compared with parental cells. The EGFR-DNR significantly reduced the ability of EGF to induce cell signaling through the MAPK pathway. Lastly, the EGFR-DNR can partially reverse cisplatin resistance in drug-resistant cells. The EGFR-DNR approach suggests that EGFR confers a growth advantage to NuTu-19 cells in vivo. Thus, EGFR blockade may ultimately prove to be a useful therapeutic tool in the treatment of cisplatin-sensitive and cisplatin-resistant ovarian cancers. PMID- 15833857 TI - Mouse p10, an alternative spliced form of p15INK4b, inhibits cell cycle progression and malignant transformation. AB - The INK4 family of proteins negatively regulates cell cycle progression at the G(1)-S transition by inhibiting cyclin-dependent kinases. Two of these cell cycle inhibitors, p16(INK4A) and p15(INK4B), have tumor suppressor activities and are inactivated in human cancer. Interestingly, both INK4 genes express alternative splicing variants. In addition to p16(INK4A), the INK4A locus encodes a splice variant, termed p12--specifically expressed in human pancreas--and ARF, a protein encoded by an alternative reading frame that acts as a tumor suppressor through the p53 pathway. Similarly, the human INK4B locus encodes the p15(INK4B) tumor suppressor and one alternatively spliced form, termed as p10. We show here that p10, which arises from the use of an alternative splice donor site within intron 1, is conserved in the mouse genome and is widely expressed in mouse tissues. Similarly to mouse p15(INK4B), p10 expression is also induced by oncogenic insults and transforming growth factor-beta treatment and acts as a cell cycle inhibitor. Importantly, we show that mouse p10 is able to induce cell cycle arrest in a p53-dependent manner. We also show that mouse p10 is able to inhibit foci formation and anchorage-independent growth in wild-type mouse embryonic fibroblasts, and that these antitransforming properties of mouse p10 are also p53 dependent. These results indicate that the INK4B locus, similarly to INK4A-ARF, harbors two different splicing variants that can be involved in the regulation of both the p53 and retinoblastoma pathways, the two major molecular pathways in tumor suppression. PMID- 15833858 TI - A hypoxia-independent hypoxia-inducible factor-1 activation pathway induced by phosphatidylinositol-3 kinase/Akt in HER2 overexpressing cells. AB - HER2 overexpression, a known prognostic factor in many human cancers, can activate phosphatidylinositol-3 kinase (PI-3K)/Akt pathways and plays an important role in mediating cell survival and tumor development. Hypoxia inducible factors (HIFs) promote angiogenesis and energy metabolism and thereby enhance tumor growth and metastasis. HIFs, composed of alpha and beta subunits, are activated in most human cancers, including those that overexpress HER2. Previous reports have suggested that increased PI-3K/Akt or decreased PTEN activity may activate the HIF pathway in various tumors, but the detailed mechanism is still not completely understood. Here we reported an interaction between the HIF and PI-3K/Akt pathways in HER2-overexpressing cancer cells. Our results indicate that HER2 overexpression, which results in constitutively active Akt, turns on HIF-1alpha independently of hypoxia, and this activation is weaker than that under hypoxic condition. Further investigation showed that Akt is required for the hypoxia-independent HIF activity. The PI-3K/Akt pathway did not affect the HIF-1alpha binding with its E3 ligase von Hippel-Lindau but enhanced the binding affinity between the HIF-1 alpha and beta subunits. Furthermore, we found that Akt interacts with HIF-1beta and regulates HIF activity. Our results indicated that HER2 can induce HIF activation via the activation of Akt suggesting that activation of HER2/Akt pathway may promote angiogenesis independent of hypoxia, which may have important implications for the oncogenic activity of HER2 and Akt. PMID- 15833859 TI - BCR-ABL induces the expression of Skp2 through the PI3K pathway to promote p27Kip1 degradation and proliferation of chronic myelogenous leukemia cells. AB - Chronic myelogenous leukemia (CML) is characterized by the expression of the BCR ABL tyrosine kinase, which results in increased cell proliferation and inhibition of apoptosis. In this study, we show in both BCR-ABL cells (Mo7e-p210 and BaF/3 p210) and primary CML CD34+ cells that STI571 inhibition of BCR-ABL tyrosine kinase activity results in a G(1) cell cycle arrest mediated by the PI3K pathway. This arrest is associated with a nuclear accumulation of p27(Kip1) and down regulation of cyclins D and E. As a result, there is a reduction of the cyclin E/Cdk2 kinase activity and of the retinoblastoma protein phosphorylation. By quantitative reverse transcription-PCR we show that BCR-ABL/PI3K regulates the expression of p27(Kip1) at the level of transcription. We further show that BCR ABL also regulates p27(Kip1) protein levels by increasing its degradation by the proteasome. This degradation depends on the ubiquitinylation of p27(Kip1) by Skp2 containing SFC complexes: silencing the expression of Skp2 with a small interfering RNA results in the accumulation of p27(Kip1). We also demonstrate that BCR-ABL cells show transcriptional up-regulation of Skp2. Finally, expression of a p27(Kip1) mutant unable of being recognized by Skp2 results in inhibition of proliferation of BCR-ABL cells, indicating that the degradation of p27(Kip1) contributes to the pathogenesis of CML. In conclusion, these results suggest that BCR-ABL regulates cell cycle in CML cells at least in part by inducing proteasome-mediated degradation of the cell cycle inhibitor p27(Kip1) and provide a rationale for the use of inhibitors of the proteasome in patients with BCR-ABL leukemias. PMID- 15833860 TI - Cortactin overexpression inhibits ligand-induced down-regulation of the epidermal growth factor receptor. AB - Ligand-induced receptor down-regulation by endocytosis is a critical process regulating the intensity and duration of receptor tyrosine kinase signaling. Ubiquitylation of specific receptor tyrosine kinases, for example, the epidermal growth factor receptor (EGFR) by the E3 ubiquitin ligase c-Cbl, provides a sorting signal for lysosomal degradation and leads to termination of receptor signaling. Cortactin, which couples the endocytic machinery to dynamic actin networks, is encoded by EMS1, a gene commonly amplified in breast and head and neck cancers. One mechanism whereby cortactin overexpression contributes to tumor progression is by enhancing tumor cell invasion and metastasis. However, in this study, we show that overexpression of cortactin in HeLa cells markedly inhibits ligand-induced down-regulation of the EGFR. This is independent of alterations in receptor autophosphorylation and correlates with impaired c-Cbl phosphorylation and association with the EGFR, reduced EGFR ubiquitylation, and sustained EGF induced extracellular signal-regulated kinase activation. Furthermore, analysis of a panel of head and neck squamous cell carcinoma (HNSCC) cell lines revealed that cortactin overexpression is associated with attenuated ligand-induced EGFR down-regulation. Importantly, RNAi-mediated reduction of cortactin expression in an 11q13-amplified HNSCC cell line accelerates EGFR degradation. This represents the first demonstration of modulation of growth factor receptor signaling by cortactin. Moreover, enhanced EGFR signaling due to cortactin overexpression may provide an alternative explanation for EMS1 gene amplification in human cancers. PMID- 15833861 TI - Role for the nuclear factor kappaB pathway in transforming growth factor-beta1 production in idiopathic myelofibrosis: possible relationship with FK506 binding protein 51 overexpression. AB - The release of transforming growth factor-beta1 (TGF-beta1) in the bone marrow microenvironment is one of the main mechanisms leading to myelofibrosis in murine models and probably in the human idiopathic myelofibrosis (IMF). The regulation of TGF-beta1 synthesis is poorly known but seems regulated by nuclear factor kappaB (NF-kappaB). We previously described the overexpression of an immunophilin, FK506 binding protein 51 (FKBP51), in IMF megakaryocytes. Gel shift and gene assays show that FKBP51's overexpression in a factor-dependent hematopoietic cell line, induces a sustained NF-kappaB activation after cytokine deprivation. This activation correlates with a low level of IkappaBalpha. A spontaneous activation of NF-kappaB was also detected in proliferating megakaryocytes and in circulating CD34(+) patient cells. In normal cells, NF kappaB activation was only detected after cytokine treatment. The expression of an NF-kappaB superrepressor in FKBP51 overexpressing cells and in derived megakaryocytes from CD34(+) of IMF patients revealed that NF-kappaB activation was not involved in the resistance to apoptosis after cytokine deprivation of these cells but in TGF-beta1 secretion. These results highlight the importance of NF-kappaB's activation in the fibrosis development of this disease. They also suggest that FKBP51's overexpression in IMF cells could play an important role in the pathogenesis of this myeloproliferative disorder. PMID- 15833862 TI - Defective p38 mitogen-activated protein kinase signaling impairs chemotaxic but not proliferative responses to stromal-derived factor-1alpha in acute lymphoblastic leukemia. AB - The chemokine stromal-derived factor-1alpha (SDF-1alpha) regulates leukemic cell motility and proliferation; however, the importance of these functions in the growth and dissemination of leukemia is unclear. We examined SDF-1alpha-mediated responses of cells from 27 cases of acute lymphoblastic leukemia (ALL). Although cells from the majority of cases showed chemotactic and proliferative responses to SDF-1alpha, a subset of cases did not undergo chemotaxis in response to SDF 1alpha, while still demonstrating dependence on SDF-1alpha for proliferation in stroma-supported cultures. This chemotactic defect was associated with an absence of phosphorylation of p38 mitogen-activated protein kinase (MAPK) induced by SDF 1alpha, and of SDF-1alpha-induced augmentation of beta(1) integrin-mediated adhesion. Signaling through phosphoinositide 3-kinase and MEK was not affected. No correlation was observed between CXCR4 expression and chemotactic function, in vitro migration into bone marrow stromal layers, and engraftment of leukemic cells in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. This study suggests that signaling through p38 MAPK is required for ALL cell chemotaxis but not for proliferation, and that the loss of a chemotactic response to SDF-1alpha does not impede engraftment in NOD/SCID mice. PMID- 15833863 TI - Differential gene up-regulation by hypoxia-inducible factor-1alpha and hypoxia inducible factor-2alpha in HEK293T cells. AB - Cells exposed to hypoxia respond by increasing the level of hypoxia-inducible factor-1 (HIF-1). This factor then activates a number of genes by binding to hypoxia response elements in their promoter regions. A second hypoxia-responsive factor, HIF-2, can activate many of the same genes as HIF-1. Overexpression of HIFs accompanies the pathogenesis of many tumors. It is unclear, however, as to the respective role of these factors in responsiveness to hypoxia and other stresses. To address this issue, we used microarray technology to study the genes activated in HEK293T cells by hypoxia or transfection with the alpha chain of HIF 1 (or mutant HIF-1 resistant to degradation) or HIF-2. Fifty-six genes were found to be up-regulated at least 3-fold by either hypoxia or transfection. Of these, 21 were elevated both by transfection with HIF-1alpha and with HIF-2alpha, and 14 were preferentially activated by HIF-1alpha including several involved in glycolysis. Ten genes were preferentially activated by HIF-2alpha, including two (CACNA1A and PTPRZ1) implicated in neurologic diseases. Interestingly, most HIF 2alpha-responsive genes were not substantially activated by hypoxia. An additional 10 genes were up-regulated by hypoxia but minimally activated by HIF 1alpha or HIF-2alpha transfection. Ten of the genes were studied by quantitative real-time PCR and/or by Northern blot and the results paralleled those found with microarray technology. Although confirmation in other systems will be necessary, these results indicate that whereas some genes are robustly activated by both HIF 1 and HIF-2, others can be preferentially activated by one or the other factor. PMID- 15833864 TI - Human bone marrow-derived mesenchymal stem cells in the treatment of gliomas. AB - The poor survival of patients with human malignant gliomas relates partly to the inability to deliver therapeutic agents to the tumor. Because it has been suggested that circulating bone marrow-derived stem cells can be recruited into solid organs in response to tissue stresses, we hypothesized that human bone marrow-derived mesenchymal stem cells (hMSC) may have a tropism for brain tumors and thus could be used as delivery vehicles for glioma therapy. To test this, we isolated hMSCs from bone marrow of normal volunteers, fluorescently labeled the cells, and injected them into the carotid artery of mice bearing human glioma intracranial xenografts (U87, U251, and LN229). hMSCs were seen exclusively within the brain tumors regardless of whether the cells were injected into the ipsilateral or contralateral carotid artery. In contrast, intracarotid injections of fibroblasts or U87 glioma cells resulted in widespread distribution of delivered cells without tumor specificity. To assess the potential of hMSCs to track human gliomas, we injected hMSCs directly into the cerebral hemisphere opposite an established human glioma and showed that the hMSCs were capable of migrating into the xenograft in vivo. Likewise, in vitro Matrigel invasion assays showed that conditioned medium from gliomas, but not from fibroblasts or astrocytes, supported the migration of hMSCs and that platelet-derived growth factor, epidermal growth factor, or stromal cell-derived factor-1alpha, but not basic fibroblast growth factor or vascular endothelial growth factor, enhanced hMSC migration. To test the potential of hMSCs to deliver a therapeutic agent, hMSCs were engineered to release IFN-beta (hMSC-IFN-beta). In vitro coculture and Transwell experiments showed the efficacy of hMSC-IFN-beta against human gliomas. In vivo experiments showed that treatment of human U87 intracranial glioma xenografts with hMSC-IFN-beta significantly increase animal survival compared with controls (P < 0.05). We conclude that hMSCs can integrate into human gliomas after intravascular or local delivery, that this engraftment may be mediated by growth factors, and that this tropism of hMSCs for human gliomas can be exploited to therapeutic advantage. PMID- 15833865 TI - Mitochondrial targeting of human O6-methylguanine DNA methyltransferase protects against cell killing by chemotherapeutic alkylating agents. AB - DNA repair capacity of eukaryotic cells has been studied extensively in recent years. Mammalian cells have been engineered to overexpress recombinant nuclear DNA repair proteins from ectopic genes to assess the impact of increased DNA repair capacity on genome stability. This approach has been used in this study to specifically target O(6)-methylguanine DNA methyltransferase (MGMT) to the mitochondria and examine its impact on cell survival after exposure to DNA alkylating agents. Survival of human hematopoietic cell lines and primary hematopoietic CD34(+) committed progenitor cells was monitored because the baseline repair capacity for alkylation-induced DNA damage is typically low due to insufficient expression of MGMT. Increased DNA repair capacity was observed when K562 cells were transfected with nuclear-targeted MGMT (nucl-MGMT) or mitochondrial-targeted MGMT (mito-MGMT). Furthermore, overexpression of mito-MGMT provided greater resistance to cell killing by 1,3-bis (2-chloroethyl)-1 nitrosourea (BCNU) than overexpression of nucl-MGMT. Simultaneous overexpression of mito-MGMT and nucl-MGMT did not enhance the resistance provided by mito-MGMT alone. Overexpression of either mito-MGMT or nucl-MGMT also conferred a similar level of resistance to methyl methanesulfonate (MMS) and temozolomide (TMZ) but simultaneous overexpression in both cellular compartments was neither additive nor synergistic. When human CD34(+) cells were infected with oncoretroviral vectors that targeted O(6)-benzylguanine (6BG)-resistant MGMT (MGMT(P140K)) to the nucleus or the mitochondria, committed progenitors derived from infected cells were resistant to 6BG/BCNU or 6BG/TMZ. These studies indicate that mitochondrial or nuclear targeting of MGMT protects hematopoietic cells against cell killing by BCNU, TMZ, and MMS, which is consistent with the possibility that mitochondrial DNA damage and nuclear DNA damage contribute equally to alkylating agent-induced cell killing during chemotherapy. PMID- 15833866 TI - Mechanisms of enhanced radiation response following epidermal growth factor receptor signaling inhibition by erlotinib (Tarceva). AB - Erlotinib (Tarceva) is an orally available HER1 (epidermal growth factor receptor, EGFR) tyrosine kinase inhibitor advancing through clinical trials for the treatment of a range of human malignancies. In this study, we examine the capacity of erlotinib to modulate radiation response and investigate specific mechanisms underlying these interactions in human tumor cell lines and xenografts. The impact of erlotinib on cell cycle kinetics was analyzed using flow cytometry, and the impact on apoptosis was evaluated via fluorescein-labeled pan-caspase inhibition and poly(ADP-ribose) polymerase cleavage. Radiation induced EGFR autophosphorylation and Rad51 expression were examined by Western blot analysis. Radiation survival was analyzed using a clonogenic assay and assessment of in vivo tumor growth was done using a mouse xenograft model system. Microarray studies were carried out using 20 K human cDNA microarray and select genes were validated using quantitative reverse transcription-PCR (RT-PCR). Independently, erlotinib and radiation induce accumulation of tumor cells in G(1) and G(2)-M phase, respectively, with a reduction of cells in S phase. When combined with radiation, erlotinib promotes a further reduction in S-phase fraction. Erlotinib enhances the induction of apoptosis, inhibits EGFR autophosphorylation and Rad51 expression following radiation exposure, and promotes an increase in radiosensitivity. Tumor xenograft studies confirm that systemic administration of erlotinib results in profound tumor growth inhibition when combined with radiation. cDNA microarray analysis assessing genes differentially regulated by erlotinib following radiation exposure identifies a diverse set of genes deriving from several functional classes. Validation is confirmed for several specific genes that may influence radiosensitization by erlotinib including Egr-1, CXCL1, and IL-1beta. These results identify the capacity of erlotinib to enhance radiation response at several levels, including cell cycle arrest, apoptosis induction, accelerated cellular repopulation, and DNA damage repair. Preliminary microarray data suggests additional mechanisms underlying the complex interaction between EGFR signaling and radiation response. These data suggest that the erlotinib/radiation combination represents a strategy worthy of further examination in clinical trials. PMID- 15833867 TI - Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. AB - The mammalian target of rapamycin (mTOR) is a downstream effector of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway and a central modulator of cell proliferation in malignant gliomas. Therefore, the targeting of mTOR signaling is considered a promising therapy for malignant gliomas. However, the mechanisms underlying the cytotoxic effects of a selective mTOR inhibitor, rapamycin, on malignant glioma cells are poorly understood. The purpose of this study was thus to elucidate how rapamycin exerts its cytotoxic effects on malignant glioma cells. We showed that rapamycin induced autophagy but not apoptosis in rapamycin-sensitive malignant glioma U87-MG and T98G cells by inhibiting the function of mTOR. In contrast, in rapamycin-resistant U373-MG cells, the inhibitory effect of rapamycin was minor, although the phosphorylation of p70S6 kinase, a molecule downstream of mTOR, was remarkably inhibited. Interestingly, a PI3K inhibitor, LY294002, and an Akt inhibitor, UCN-01 (7 hydroxystaurosporine), both synergistically sensitized U87-MG and T98G cells as well as U373-MG cells to rapamycin by stimulating the induction of autophagy. Enforced expression of active Akt in tumor cells suppressed the combined effects of LY294002 or UCN-01, whereas dominant-negative Akt expression was sufficient to increase the sensitivity of tumor cells to rapamycin. These results indicate that rapamycin exerts its antitumor effect on malignant glioma cells by inducing autophagy and suggest that in malignant glioma cells a disruption of the PI3K/Akt signaling pathway could greatly enhance the effectiveness of mTOR inhibitors. PMID- 15833868 TI - Construction of a minimal HIV-1 variant that selectively replicates in leukemic derived T-cell lines: towards a new virotherapy approach. AB - T-cell acute lymphoblastic leukemia is a high-risk type of blood-cell cancer. We analyzed the possibility of developing virotherapy for T-cell acute lymphoblastic leukemia. Virotherapy is based on the exclusive replication of a virus in leukemic cells, leading to the selective removal of these malignant cells. We constructed a minimized derivative of HIV-1, a complex lentivirus encoding multiple accessory functions that are essential for virus replication in untransformed cells, but dispensable in leukemic T cells. This mini-HIV virus has five deletions (vif, vpR, vpU, nef, and U3) and replicated in the SupT1 cell line, but did not replicate in normal peripheral blood mononuclear cells. The stripped down mini-HIV variant was also able to efficiently remove leukemic cells from a mixed culture with untransformed control cells. In contrast to wild-type HIV-1, we did not observe bystander killing in mixed culture experiments with the mini-HIV variant. Furthermore, viral escape was not detected in long-term cultures. The mini-HIV variant that uses CD4 and CXCR4 for cell entry could potentially be used against CXCR4-expressing malignancies such as T-lymphoblastic leukemia/lymphoma, natural killer leukemia, and some myeloid leukemias. PMID- 15833869 TI - Magnetic resonance spectroscopy monitoring of mitogen-activated protein kinase signaling inhibition. AB - Several mitogen-activated protein kinase (MAPK) signaling inhibitors are currently undergoing clinical trial as part of novel mechanism-based anticancer treatment strategies. This study was aimed at detecting biomarkers of MAPK signaling inhibition in human breast and colon carcinoma cells using magnetic resonance spectroscopy. We investigated the effect of the prototype MAPK kinase inhibitor U0126 on the (31)P-MR spectra of MDA-MB-231, MCF-7 and Hs578T breast, and HCT116 colon carcinoma cells. Treatment of MDA-MB-231 cells with 50 micromol/L U0126 for 2, 4, 8, 16, 24, 32, and 40 hours caused inhibition of extracellular signal-regulated kinases (ERK1/2) phosphorylation from 2 hours onwards. (31)P-MR spectra of extracted cells indicated that this was associated with a significant drop in phosphocholine levels to 78 +/- 8% at 8 hours, 74 +/- 8% at 16 hours, 66 +/- 7% at 24 hours, 71 +/- 10% at 32 hours, and 65 +/- 10% at 40 hours post-treatment. In contrast, the lower concentration of 10 micromol/L U0126 for 40 hours had no significant effect on either P-ERK1/ 2 or phosphocholine levels in MDA-MB-231 cells. Depletion of P-ERK1/2 in MCF-7 and Hs578T cells with 50 micromol/L U0126 also produced a drop in phosphocholine levels to 51 +/- 17% at 40 hours and 23 +/- 12% at 48 hours, respectively. Similarly, in HCT116 cells, inhibition with 30 micromol/L U0126 caused depletion of P-ERK1/2 and a decrease in phosphocholine levels to 80 +/- 9% at 16 hours and 61 +/- 4% at 24 hours post-treatment. The reduction in phosphocholine in MDA-MB 231 and HCT116 cells correlated positively with the drop in P-ERK1/2 levels. Our results show that MAPK signaling inhibition with U0126 is associated with a time dependent decrease in cellular phosphocholine levels. Thus, phosphocholine has potential as a noninvasive pharmacodynamic marker for monitoring MAPK signaling blockade. PMID- 15833870 TI - Phenoxodiol, a novel isoflavone, induces G1 arrest by specific loss in cyclin dependent kinase 2 activity by p53-independent induction of p21WAF1/CIP1. AB - Phenoxodiol, an isoflavone derivative of genistein with unknown mechanism of action, is currently being evaluated in early human cancer clinical trials. To determine the mechanism of antiproliferative effects of phenoxodiol, we examined its effects in a battery of human cell lines. Although we observed caspase dependent apoptosis in HN12 cells as early as 24 hours after exposure, clonogenic death occurred only after 48-hour exposure despite caspase blockade by the general caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (ZVAD)-fmk. Moreover, clear evidence of cell death as determined by nuclear morphology and plasmatic membrane damage occur despite ZVAD, suggesting that another mechanism besides caspase-dependent apoptosis is required for clonogenic death induced by phenoxodiol. In search for other potential antiproliferative effects, we assessed the effects of phenoxodiol in the cell cycle progression of human carcinoma cell lines. A significant G(1)-S arrest was observed by 12 hours of exposure in HN12 cell lines at concentrations > or =5 microg/mL. Cell cycle arrest occurred several hours (approximately 12 hours) before induction of apoptosis. Analysis of in vitro purified cyclin-dependent kinase (cdk) activity showed that phenoxodiol did not inhibit cdk activity. In contrast, cellular cdk2 activity obtained from HN12 cell lines exposed to phenoxodiol for 12 hours decreased by 60%, whereas cdk6 activity remained unaltered, suggesting that the loss of cdk2 activity was specific. Loss in cdk2 activity was preceded by the accumulation of the endogenous cdk inhibitor p21(WAF1). To assess the role of p21(WAF1) induction by phenoxodiol, we used HCT116 isogenic cell lines and showed that phenoxodiol induced G(1) arrest together with p21(WAF1) expression in wild type clones. In contrast, p21(-/-) variants failed to show G(1) arrest. Finally, induction of p21 by phenoxodiol is p53 independent, as phenoxodiol induced p21 in HCT116 lacking p53. These data therefore indicate that phenoxodiol promotes G(1) S arrest by the specific loss in cdk2 activity due to p53-independent p21(WAF1) induction. This novel feature of phenoxodiol may have clinical implications, as the majority of human malignancies have aberrations in cell cycle progression regulation. PMID- 15833871 TI - C-C chemokine receptor 5 on stromal cells promotes pulmonary metastasis. AB - We have shown that mice that express the C-C chemokine receptor 5 (CCR5) have enhanced local tumor growth and an impaired response to vaccine therapy compared with CCR5 knockout (CCR5(-/-)) mice. Here, we extend these observations to evaluate the function of CCR5 in pulmonary metastasis and the mechanism underlying the diminished tumor growth in CCR5(-/-) mice. Lung metastases were counted in wild-type (WT) and CCR5(-/-) mice following the injection of 1 x 10(6) B16-F10 melanoma cells. These results were compared with those from syngeneic bone marrow chimeric mice formed by the transfer of WT bone marrow into irradiated CCR5(-/-) and CCR5(-/-) marrow into irradiated WT mice. Intact CCR5(-/ ) mice developed fewer metastases than WT mice (40.2 versus 70.6; P < 0.05). Bone marrow chimeras formed by the transfer of WT bone marrow into CCR5(-/-) hosts had fewer metastases than WT hosts injected with knockout marrow (46.6 versus 98.6; P < 0.01). Adoptive transfer of CCR5-expressing leukocytes also failed to promote metastasis in CCR5(-/-) mice. However, the i.v. transfer of WT pulmonary stromal cells into CCR5(-/-) mice increased the number of metastases compared with transfer of CCR5(-/-) stromal cells (102.8 versus 26.0; P < 0.05). These results show for the first time that CCR5 expression on stromal and not hematopoietic cells contributes to tumor metastasis. Therefore, recently developed CCR5 inhibitors may have a novel benefit in cancer therapy. PMID- 15833872 TI - Target cell-restricted apoptosis induction of acute leukemic T cells by a recombinant tumor necrosis factor-related apoptosis-inducing ligand fusion protein with specificity for human CD7. AB - Current treatment of human T-cell leukemia and lymphoma is predominantly limited to conventional cytotoxic therapy and is associated with limited therapeutic response and significant morbidity. Therefore, more potent and leukemia-specific therapies with favorable toxicity profiles are urgently needed. Here, we report on the construction of a novel therapeutic fusion protein, scFvCD7:sTRAIL, designed to induce target antigen-restricted apoptosis in human T-cell tumors. ScFvCD7:sTRAIL consists of the death-inducing tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) genetically linked to an scFv antibody fragment specific for the T-cell surface antigen CD7. Treatment with scFvCD7:sTRAIL induced potent CD7-restricted apoptosis in a series of malignant T-cell lines, whereas normal resting leukocytes, activated T cells, and vascular endothelial cells (human umbilical vein endothelial cells) showed no detectable apoptosis. The apoptosis-inducing activity of scFvCD7:sTRAIL was stronger than that of the immunotoxin scFvCD7:ETA. In mixed culture experiments with CD7-positive and CD7 negative tumor cells, scFvCD7:sTRAIL induced very potent bystander apoptosis of CD7-negative tumor cells. In vitro treatment of blood cells freshly derived from T-acute lymphoblastic leukemia patients resulted in marked apoptosis of the malignant T cells that was strongly augmented by vincristin. In conclusion, scFvCD7:sTRAIL is a novel recombinant protein causing restricted apoptosis in human leukemic T cells with low toxicity for normal human blood and endothelial cells. PMID- 15833873 TI - Anticancer activity of the antibiotic clioquinol. AB - Clioquinol, a metal chelator, has been used for many years as an antimicrobial agent and more recently as a potential treatment for Alzheimer's disease. Because it binds copper and zinc, metals essential for the activity of the enzyme superoxide dismutase-1 (SOD1), a potential target for anticancer drug development, we investigated its effects on human cancer cells. Treatment with clioquinol reduced the viability of eight different human cancer cell lines in a concentration-dependent manner, with IC(50) values in the low micromolar range. Biochemical analysis revealed that clioquinol induced cancer cell death through apoptotic pathways that require caspase activity. Although clioquinol induced modest inhibition of SOD1 activity in treated cells, comparable inhibition by a known SOD1 inhibitor, diethyldithiocarbamate, did not result in cytotoxicity. The addition of copper, iron, or zinc did not rescue cells from cliquinol-induced cytotoxicity but enhanced its killing, arguing against metal chelation as its major mechanism of action. To test if clioquinol might act as an ionophore, a fluorescent probe was used to monitor intracellular zinc concentrations. The addition of clioquinol resulted in elevated levels of intracellular zinc, indicating that clioquinol acts as a zinc ionophore. In an in vivo xenografts mouse model, clioquinol inhibited tumor growth of xenografts over a 6-week period, without inducing visible toxicity. Our results show that clioquinol has anticancer effects both in vitro and in vivo. Transition metal ionophores may be a subclass of metal chelators with anticancer activity deserving of further development. PMID- 15833874 TI - Dietary genistein reduces metastasis in a postsurgical orthotopic breast cancer model. AB - Metastatic spread, not primary tumor burden, is the leading cause of breast cancer deaths. For patient prognosis to improve, new systemic adjuvant therapies that are capable of effectively inhibiting the outgrowth of seeded tumor cells after surgical treatment of the primary breast tumor are needed. To facilitate the preclinical development of such therapies, relevant animal models of breast cancer metastasis that can mimic the postsurgical adjuvant setting are required. Here we developed a preclinical xenograft model of breast cancer metastasis where the primary tumor was removed by surgical resection before systemic adjuvant treatment. We used this model to assess the antimetastatic effect of postsurgical dietary intervention with the soy isoflavone genistein. The anticancer activity of genistein has been established in vitro and in vivo, however, few studies have tested the potential of genistein as an antimetastatic therapy. Using our model, we tested the efficacy of adjuvant treatment with genistein to inhibit the outgrowth of metastases postsurgery. To establish primary tumors, human breast carcinoma cells, MDA-MB-435/HAL, were implanted into the mammary fat pad of female nude mice. Primary tumors were left to grow for 5 weeks before being surgically removed. Mice were then randomized into two diet groups: control soy free diet versus genistein-supplemented diet. Five weeks later, metastatic burden was assessed. Genistein reduced the percent metastatic burden in the lungs by 10 fold. These results indicate that dietary intervention following cancer surgery can affect the outgrowth of seeded tumor cells. The availability of well characterized, clinically relevant animal models for studying factors that regulate metastatic outgrowth postsurgery will provide an important tool for developing new systemic adjuvant therapies. PMID- 15833875 TI - Inhibition of HER-2/neu kinase impairs androgen receptor recruitment to the androgen responsive enhancer. AB - Advanced prostate cancer invariably recurs despite androgen deprivation therapy. The androgen receptor (AR) likely plays a key role in this progression and in the continued survival and proliferation of prostate cancer cells in the low androgen environment. Cross-talk with growth factor receptors, such as epidermal growth factor receptor (EGFR) family, has been postulated as a potential mechanism to activate AR in recurrent prostate cancer. We have investigated the role of HER 2/neu (ErbB-2) tyrosine kinase in AR function by characterizing the effect of inhibiting endogenous HER-2 activity in LNCaP cells. We used two independent methods, expression of intracellular single-chain antibody against HER-2 and treatment with a novel dual EGFR/HER-2 kinase inhibitor GW572016 (lapatinib). Expression of intracellular HER-2 antibody scFv-5R and treatment with GW572016 inhibited HER-2 signaling. This HER-2 inhibition led to impairment of AR-mediated functions, such as androgen-stimulated growth and the induction of endogenous prostate-specific antigen (PSA) mRNA and protein. Androgen-stimulated recruitment of AR and histone acetylation at the androgen responsive enhancer of the PSA gene, detected by chromatin immunoprecipitation analysis, were impaired by HER-2 inhibition. GW572016 was more potent in its ability to inhibit PSA expression and AR recruitment and histone acetylation than the EGFR-selective kinase inhibitor ZD1839 (gefitinib), consistent with the HER-2 kinase playing the major role in AR regulation. These results show that HER-2 signaling is required for optimal transcriptional activity of AR in prostate cancer cells and suggest that HER-2 inhibition may provide a novel strategy to disrupt AR function in prostate cancer. PMID- 15833876 TI - DNA vaccine expressing the mimotope of GD2 ganglioside induces protective GD2 cross-reactive antibody responses. AB - The GD2 ganglioside expressed on neuroectodermally derived tumors, including neuroblastoma and melanoma, is weakly immunogenic in tumor-bearing patients and induces predominantly immunoglobulin (Ig)-M antibody responses in the immunized host. Here, we investigated whether interconversion of GD2 into a peptide mimetic form would induce GD2 cross-reactive IgG antibody responses in mice. Screening of the X(15) phage display peptide library with the anti-GD2 monoclonal antibody (mAb) 14G2a led to isolation of mimetic peptide 47, which inhibited the binding of 14G2a antibody to GD2-positive tumor cells. The peptide was also recognized by GD2-specific serum antibodies from a patient with neuroblastoma, suggesting that it bears an internal image of GD2 ganglioside expressed on the tumor cells. The molecular basis for antigenicity of the GD2 mimetic peptide, established by molecular modeling and mutagenesis studies, led to the generation of a 47-LDA mutant with an increased mimicry to GD2. Immunization of mice with peptide 47-LDA encoded plasmid DNA elicited GD2 cross-reactive IgG antibody responses, which were increased on subsequent boost with GD2 ganglioside. The vaccine-induced antibodies recognized GD2-positive tumor cells, mediated complement-dependent cytotoxicity, and exhibited protection against s.c. human GD2-positive melanoma growth in the severe combined immunodeficient mouse xenograft model. The results from our studies provide insights into approaches for boosting GD2 cross-reactive IgG antibody responses by minigene vaccination with a protective epitope of GD2 ganglioside. PMID- 15833877 TI - A DNA vaccine targeting Fos-related antigen 1 enhanced by IL-18 induces long lived T-cell memory against tumor recurrence. AB - A novel vaccination strategy induced specific CD8(+) T cell-mediated immunity that eradicated spontaneous and experimental pulmonary cancer metastases in syngeneic mice and was also effective in a therapeutic setting of established breast cancer metastases. This was achieved by targeting transcription factor Fos related antigen 1(Fra-1), overexpressed by many tumor cells, with an ubiquitinated DNA vaccine against Fra-1, coexpressing secretory IL-18. Insight into the immunologic mechanisms involved was provided by adoptive transfer of T lymphocytes from successfully immunized BALB/c mice to syngeneic severe combined immunodeficient (SCID) mice. Specifically, long-lived T memory cells were maintained dormant in nonlymphoid tissues by IL-18 in the absence of tumor antigen. Importantly, a second tumor cell challenge of these SCID mice restored both, robust tumor-specific cytotoxicity and long-lived T-cell memory, capable of eradicating established pulmonary cancer metastases, suggesting that this vaccine could be effective against tumor recurrence. PMID- 15833878 TI - Constitutive expression and costimulatory function of LIGHT/TNFSF14 on human melanoma cells and melanoma-derived microvesicles. AB - Neoplastic cells are thought to have defective expression of costimulatory molecules. However, in this study, we show that human melanoma cells express LIGHT/TNFSF14, a ligand of herpesvirus entry mediator on T cells and of lymphotoxin beta receptor on stromal cells. In vitro, melanoma cells stained for LIGHT in the intracellular compartment, with weak or negative cell surface expression. However, LIGHT was expressed on tumor-derived microvesicles released from melanoma cells. In vivo, LIGHT was found in metastatic lesions, and the extent of lymphotoxin beta receptor expression on the stromal cells was significantly associated with a "brisk" T-cell infiltrate in the neoplastic tissue. In the lesions with a brisk T-cell infiltrate, stromal cells surrounding the tumor also stained for the T-cell attractant chemokine CCL21. The intratumoral T lymphocytes frequently expressed herpesvirus entry mediator and were characterized by a differentiated phenotype. Coculture of lymphocytes with LIGHT(+) melanoma-derived microvesicles or even with LIGHT(+) melanoma cells in the presence of interleukin-2 costimulated LIGHT-dependent CD3(+)CD8(+) T-cell proliferation. However, lymphocyte coculture with LIGHT(+) microvesicles in the presence of interleukin-2 was also associated with an apoptotic response as documented by increased binding of Annexin V by CD3(+)CD8(+) T cells. These data suggest that LIGHT constitutively expressed in human melanoma cells and microvesicles may contribute to regulate T-cell responses to tumor cells. PMID- 15833879 TI - Redirecting in vivo elicited tumor infiltrating macrophages and dendritic cells towards tumor rejection. AB - A hostile tumor microenvironment interferes with the development and function of the adaptive immune response. Here we report the mechanisms by which large numbers of tumor-infiltrating macrophages and dendritic cells (DC) can be redirected to become potent effectors and activators of the innate and adaptive immunity, respectively. We use adenoviral delivery of the CCL16 chemokine to promote accumulation of macrophages and DC at the site of preestablished tumor nodules, combined with the Toll-like receptor 9 ligand CpG and with anti interleukin-10 receptor antibody. CpG plus anti-interleukin-10 receptor antibody promptly switched infiltrating macrophages infiltrate from M2 to M1 and triggered innate response debulking large tumors within 16 hours. Tumor-infiltrating DC matured and migrated in parallel with the onset of the innate response, allowing the triggering of adaptive immunity before the diffuse hemorrhagic necrosis halted the communication between tumor and draining lymph nodes. Treatment of B6>CXB6 chimeras implanted with BALB/c tumors with the above combination induced an efficient innate response but not CTL-mediated tumor lysis. In these mice, tumor rejection did not exceed 25%, similarly to that observed in CCR7-null mice that have DC unable to prime an adaptive response. The requirement of CD4 help was shown in CD40-KO, as well as in mice depleted of CD4 T cells, during the priming rather than the effector phase. Our data describe the critical requirements for the immunologic rejection of large tumors: a hemorrhagic necrosis initiated by activated M1 macrophages and a concomitant DC migration to draining lymph nodes for subsequent CTL priming and clearing of any tumor remnants. PMID- 15833880 TI - IFN unresponsiveness in LNCaP cells due to the lack of JAK1 gene expression. AB - We reported previously that 23% of human lung adenocarcinoma cell lines were unresponsive to IFN-gamma. To extend this finding to cancer cells derived from distinct tissues of origin, we assessed IFN-gamma receptor signaling in the LNCaP human prostate adenocarcinoma cell line, which in previous experiments by others failed to induce a range of IFN-dependent biological responses. In this report, we show that LNCaP cells fail to respond to either IFN-gamma or IFN-alpha because of an impairment in the proximal signaling events downstream of both IFN-gamma and IFN-alpha/beta receptors that lead to the activation of STAT1. Furthermore, we show that LNCaP insensitivity to the IFNs is a result of the absence of expression of the JAK1 kinase, an obligate component shared by both IFN-gamma and IFN-alpha/beta receptors. JAK1 was undetectable in LNCaP cells at both protein and message levels. Treatment of LNCaP cells with a combination of inhibitors of DNA methyltransferases and histone deacetylases induced expression of JAK1 message. These results identify the molecular basis for IFN insensitivity in the LNCaP cell line and suggest that epigenetic silencing of key immunologic signaling components may be one mechanism by which tumor cells evade immune detection and elimination. PMID- 15833881 TI - Combined genetic assessment of transforming growth factor-beta signaling pathway variants may predict breast cancer risk. AB - There is growing evidence that common variants of the transforming growth factor beta (TGF-beta) signaling pathway may modify breast cancer risk. In vitro studies have shown that some variants increase TGF-beta signaling, whereas others have an opposite effect. We tested the hypothesis that a combined genetic assessment of two well-characterized variants may predict breast cancer risk. Consecutive patients (n = 660) with breast cancer from the Memorial Sloan-Kettering Cancer Center (New York, NY) and healthy females (n = 880) from New York City were genotyped for the hypomorphic TGFBR1*6A allele and for the TGFB1 T29C variant that results in increased TGF-beta circulating levels. Cases and controls were of similar ethnicity and geographic location. Thirty percent of cases were identified as high or low TGF-beta signalers based on TGFB1 and TGFBR1 genotypes. There was a significantly higher proportion of high signalers (TGFBR1/TGFBR1 and TGFB1*CC) among controls (21.6%) than cases (15.7%; P = 0.003). The odds ratio [OR; 95% confidence interval (95% CI)] for individuals with the lowest expected TGF-beta signaling level (TGFB1*TT or TGFB1*TC and TGFBR1*6A) was 1.69 (1.08 2.66) when compared with individuals with the highest expected TGF-signaling levels. Breast cancer risk incurred by low signalers was most pronounced among women after age 50 years (OR, 2.05; 95% CI, 1.01-4.16). TGFBR1*6A was associated with a significantly increased risk for breast cancer (OR, 1.46; 95% CI, 1.04 2.06), but the TGFB1*CC genotype was not associated with any appreciable risk (OR, 0.89; 95% CI, 0.63-1.21). TGFBR1*6A effect was most pronounced among women diagnosed after age 50 years (OR, 2.20; 95% CI, 1.25-3.87). This is the first study assessing the TGF-beta signaling pathway through two common and functionally relevant TGFBR1 and TGFB1 variants. This approach may predict breast cancer risk in a large subset of the population. PMID- 15833882 TI - The retinoid X receptor-selective retinoid, LGD1069, down-regulates cyclooxygenase-2 expression in human breast cells through transcription factor crosstalk: implications for molecular-based chemoprevention. AB - Retinoids and their derivatives can suppress the development of cancer in animals and in humans. We and others have shown that retinoid X receptor (RXR)-selective retinoids or "rexinoids" suppress the development of breast cancer in several animal models with minimal toxicity. LGD1069 (Bexarotene) is a potent RXR selective retinoid with reduced toxicity compared with naturally occurring retinoids. In this study, we investigated the expression of LGD1069-modulated biomarkers. We previously did cDNA array analysis of LGD1069-treated breast cells using Affymetrix microarrays. These studies identified many LGD1069-regulated genes, one of which was cyclooxygenase-2 (COX-2). Because COX-2 inhibitors have been shown to prevent cancer in other model systems, we investigated whether LGD1069 inhibits the expression of COX-2 in mammary tissue and in normal human mammary epithelial cells (HMEC). In mouse mammary tumor virus-erbB2 mice treated with LGD1069, there was a marked decrease of COX-2 expression in both normal and malignant mammary tissues. The effect of LGD1069 on COX-2 expression was also investigated in normal human breast cells. COX-2 expression was markedly reduced by treatment with LGD1069 at the RNA and protein level in normal HMECs; LGD1069 suppressed COX-2 promoter activity. We also showed that LGD1069 inhibited activator protein (AP-1)-dependent transcription in these breast cells, and that suppression of COX-2 expression was due to sequestration of CBP/p300. These results from in vivo and in vitro studies suggest that LGD1069, an RXR-selective retinoid, inhibits COX-2 expression by suppression of COX-2 transcription in part through transrepression of the AP-1 transcription factor. Thus, RXR-selective retinoids that inhibit AP-1 activity and suppress COX-2 expression may be particularly promising drugs for breast cancer prevention. Furthermore, such RXR selective retinoids may be most useful in combination with antiestrogens for more effective prevention of breast cancer in women at high risk of this disease. PMID- 15833883 TI - Genistein inhibits p38 map kinase activation, matrix metalloproteinase type 2, and cell invasion in human prostate epithelial cells. AB - Epidemiologic studies associate consumption of genistein, in the form of dietary soy, with lower rates of metastatic prostate cancer. We have previously shown that genistein inhibits prostate cancer cell detachment in vitro, that it is well tolerated in an older cohort of men with prostate cancer, and that it alters cell signaling in that same cohort. We have also shown that p38 mitogen-activated protein kinase (MAPK) is necessary for transforming growth factor beta (TGF-beta) mediated increases in prostate cancer adhesion. Although cell invasion is closely linked to metastatic behavior, little is known about how this process is regulated in prostate cancer or what effect, if any, genistein has on associated processes. We now show that genistein inhibits matrix metalloproteinase type 2 (MMP-2) activity in six of seven prostate cell lines tested, blocks MMP-2 induction by TGF-beta, and inhibits cell invasion. Efficacy was seen at low nanomolar concentrations, corresponding to blood concentrations of free genistein attained after dietary consumption. Inhibition of p38 MAPK by either SB203580 or dominant-negative construct blocked induction of MMP-2 and cell invasion by TGF beta. Genistein exerted similar effects and was found to block activation of p38 MAPK by TGF-beta. This study shows that p38 MAPK is necessary for TGF-beta mediated induction of MMP-2 and cell invasion in prostate cancer and that genistein blocks activation of p38 MAPK, thereby inhibiting processes closely linked to metastasis, and does so at concentrations associated with dietary consumption. Any potential causal link to epidemiologic findings will require further investigation. PMID- 15833884 TI - Flavokawain A, a novel chalcone from kava extract, induces apoptosis in bladder cancer cells by involvement of Bax protein-dependent and mitochondria-dependent apoptotic pathway and suppresses tumor growth in mice. AB - Consumption of the traditional kava preparation was reported to correlate with low and uncustomary gender ratios (more cancer in women than men) of cancer incidences in three kava-drinking countries: Fiji, Vanuatu, and Western Samoa. We have identified flavokawain A, B, and C but not the major kavalactone, kawain, in kava extracts as causing strong antiproliferative and apoptotic effect in human bladder cancer cells. Flavokawain A results in a significant loss of mitochondrial membrane potential and release of cytochrome c into the cytosol in an invasive bladder cancer cell line T24. These effects of flavokawain A are accompanied by a time-dependent decrease in Bcl-x(L), a decrease in the association of Bcl-x(L) to Bax, and an increase in the active form of Bax protein. Using the primary mouse embryo fibroblasts Bax knockout and wild-type cells as well as a Bax inhibitor peptide derived from the Bax-binding domain of Ku70, we showed that Bax protein was, at least in part, required for the apoptotic effect of flavokawain A. In addition, flavokawain A down-regulates the expression of X-linked inhibitor of apoptosis and survivin. Because both X-linked inhibitor of apoptosis and survivin are main factors for apoptosis resistance and are overexpressed in bladder tumors, our data suggest that flavokawain A may have a dual efficacy in induction of apoptosis preferentially in bladder tumors. Finally, the anticarcinogenic effect of flavokawain A was evident in its inhibitory growth of bladder tumor cells in a nude mice model (57% of inhibition) and in soft agar. PMID- 15833885 TI - Selenium disrupts estrogen signaling by altering estrogen receptor expression and ligand binding in human breast cancer cells. AB - Cancer prevention studies suggest that selenium is effective in reducing the incidence of cancers including prostate, colon, and lung cancers. Previous reports showed that selenium inhibits premalignant human breast MCF-10AT1 and MCF10AT3B cell growth in vitro and reduces mammary tumor incidence after exposure to carcinogens in tumor models. Because estrogen is critical to the development and differentiation of estrogen target tissues, including the breast, the present study was designed to examine the effect of selenium on estrogen receptor (ER) expression and activation using methylseleninic acid (MSA), an active form of selenium in vitro. Selenium decreased the levels of expression of ERalpha mRNA and protein and reduced the binding of labeled estradiol to estrogen receptor in MCF-7 cells. Selenium inhibited the trans-activating activity of estrogen receptor in MCF-7 cells expressing functional estrogen receptor using a luciferase reporter construct linked to estrogen responsive element. Selenium decreased the binding of estrogen receptor to the estrogen responsive element site using an electrophoretic mobility gel shift assay. Selenium suppressed estrogen induction of the endogenous target gene c-myc. In contrast to the effect on ERalpha in MCF-7 cells, selenium increased ERbeta mRNA expression in MDA-MB231 human breast cancer cells. Thus, differential regulation of ERalpha and ERbeta in breast cancer cells may represent a novel mechanism of selenium action and provide a rationale for selenium breast cancer prevention trial. PMID- 15833886 TI - Tamoxifen inhibition of estrogen receptor-alpha-negative mouse mammary tumorigenesis. AB - Tamoxifen reduces the relative risk of breast cancer developing from specific premalignant lesions. Many breast cancers that arise after tamoxifen treatment are estrogen receptor-alpha (ER-alpha)-negative, although premalignant lesions such as atypical ductal hyperplasia are highly ER-alpha-positive. The p53 null mouse mammary epithelial transplant model is characterized by ER-alpha-positive premalignant lesions that give rise to both ER-alpha-positive and ER-alpha negative tumors. Given this progression from ER-alpha-positive to ER-alpha negative lesions, we tested the ability of tamoxifen to block or delay mammary tumorigenesis in several versions of this model. In groups 1 and 2, p53 null normal mammary epithelial transplants were maintained in virgin mice. In groups 3 to 5, the p53 null and mammary transplants were maintained in mice continuously exposed to high levels of progesterone. In groups 6 and 7, transplants of the premalignant outgrowth line PN8a were maintained in virgin mice. Tamoxifen blocked estrogen signaling in these mice as evidenced by decreases in progesterone-induced lateral branching and epithelial proliferation in the mammary epithelium. Tamoxifen did not alter the elevated levels of progesterone in the blood while significantly reducing the circulating level of prolactin. Tamoxifen reduced tumor incidence in p53 null normal mammary epithelial transplants maintained in virgin mice from 55% to 5% and in progesterone stimulated mice from 81% to 21%. The majority of the resultant tumors were ER alpha-negative. Tamoxifen also significantly delayed tumorigenesis in the ER alpha-positive high premalignant line PN8a from 100% to 75%. These results show that tamoxifen delays the emergence of ER-alpha-negative tumors if given early in premalignant progression. PMID- 15833887 TI - Fluticasone inhalation in moderate cases of bronchopulmonary dysplasia. AB - OBJECTIVE: This randomized, controlled trial was designed to determine the efficacy of inhaled fluticasone propionate on oxygen therapy weaning in a population of preterm infants who were born at <32 weeks of gestation and experienced moderate bronchopulmonary dysplasia (BPD). METHODS: Thirty-two infants who were < or =32 weeks of gestation, had moderate BPD that required supplemental oxygen (fraction of inspired oxygen > or =0.25), and were aged between 28 and 60 days were randomized. Fluticasone propionate 125 microg twice daily for 3 weeks and once daily for a fourth week was delivered to infants who weighed between 500 and 1200 g. The dosage was doubled for infants who weighed > or =1200 g. RESULTS: Compared with placebo, treatment had no effect on either duration of supplemental O2 therapy or ventilatory support as assessed by survival analysis. At 28 days, a trend toward a lower cortisol/creatinine ratio in the treatment group was noted compared with placebo (25.1 +/- 18.9 vs 43 +/- 14.4). In the fluticasone group at 28 days, the systolic arterial pressure (78 +/ 3 vs 68 +/- 3 mm Hg) and diastolic arterial pressure (43 +/- 3.4 mm Hg vs 38 +/- 2.0 mm Hg) were higher compared with baseline fluticasone values. The chest radiograph score was lower than baseline (2.8 +/- 1.4 vs 3.7 +/- 2.2) in the fluticasone group at 28 days. This study has a statistical power of 1.0 to detect a significant difference in the duration of oxygen supplementation of >21 days between the study groups. CONCLUSION: We conclude that fluticasone propionate reduces neither supplemental O2 use nor the need for ventilatory support in this patient population. However, fluticasone does have a positive radiologic effect in lowering chest radiograph scores. In addition, our data point to a possible association among inhaled fluticasone treatment and higher arterial blood pressure. Thus, the results of this investigation do not support the use of inhaled corticosteroids in the treatment of oxygen-dependent infants who have established moderate BPD. PMID- 15833888 TI - NEMO mutations in 2 unrelated boys with severe infections and conical teeth. AB - X-linked recessive anhidrotic ectodermal dysplasia with immunodeficiency is a developmental and immunologic disorder caused by mutations in nuclear factor kappaB essential modulator (NEMO), which is essential for nuclear factor-kappaB activation. Early in life, affected boys present a typical appearance, with hypotrichosis or atrichosis, hypohidrosis or anhidrosis, and hypodontia or anodontia with conical incisors. They are also susceptible to various microorganisms, mostly pyogenic bacteria and mycobacteria. Here we report 2 unrelated boys, aged 6 and 11 years, who have novel mutations in NEMO and present conical incisors and hypodontia as their sole and long-unrecognized developmental anomaly. One child had isolated recurrent pneumococcal disease, whereas the other had multiple infections. Our observations indicate that conical incisors should prompt the search for NEMO mutations in boys with unusual infectious diseases. PMID- 15833889 TI - Central hypothyroidism in infants who were born to mothers with thyrotoxicosis before 32 weeks' gestation: 3 cases. AB - We describe 3 infants who were born to mothers with Graves' disease and developed central hypothyroidism that persisted for >6 months after birth. Two were preterm infants, and the other was a term infant who was born to a euthyroid mother who had been treated with an antithyroid drug since week 31 of gestation. These cases suggest that passage of thyroid hormones can occur from a thyrotoxic mother to the fetus and that the gestational period earlier than 32 weeks may be the critical time for development of central hypothyroidism. PMID- 15833890 TI - Respiratory failure and hypercoagulability in a toddler with Lemierre's syndrome. AB - A 3.5-year-old healthy boy with 4 days of fever was referred to the emergency department for respiratory distress. The physical examination was remarkable for stupor, tachycardia, tachypnea, and dyspnea. Initial blood tests showed pancytopenia. He rapidly developed torticollis. Computerized tomography of the neck revealed a thrombus in the internal jugular vein. A presumptive diagnosis of Lemierre's syndrome was made and he was started on antibiotics and anticoagulation. He subsequently developed adult respiratory distress syndrome and required high frequency oscillatory ventilation for 9 days. Blood cultures were positive for Fusobacterium necrophorum. Screening for hypercoagulability revealed 2 known risk factors: a mutation in the prothrombin gene and elevated lipoprotein a. PMID- 15833891 TI - Roles for nicotinic acetylcholine receptor subunit large cytoplasmic loop sequences in receptor expression and function. AB - To evaluate possible physiological roles of the large cytoplasmic loops (C2) and neighboring transmembrane domains of nicotinic acetylcholine receptor (nAChR) subunits, we generated novel fusion constructs in which human nAChR alpha4, beta2, or beta4 subunit C2 or C2 and neighboring sequences were replaced by corresponding sequences from the mouse serotonin type 3A (5-HT3A) receptor subunit. Following stable expression in human SH-EP1 cells, we found that extensive sequence substitutions involving third and fourth transmembrane domains and neighboring "proximal" C2 sequences (e.g., beta2 H322-V335 and V449-R460) did not allow functional expression of nAChR containing chimeric subunits. However, expression of functional nAChR was achieved containing wild-type alpha4 subunits and chimeric beta2 (beta2chi) subunits whose "nested" C2 domain sequences K336 S448 were replaced with the corresponding 5-HT3A subunit sequences. Whereas these findings suggested indispensable roles for M3/M4 transmembrane and/or proximal C2 sequences in alpha4beta2-nAChR function, nested C2 sequences in the beta2 subunit are not essential for functional receptor expression. Ligand-binding analyses also revealed only subtle differences in pharmacological profiles of alpha4beta2 nAChR compared with alpha4beta2chi-nAChR. Nevertheless, there was heightened emergence of agonist-mediated self-inhibition of alpha4beta2chi function, greater sensitivity to functional blockade by a number of antagonists, and faster and more complete acute desensitization of alpha4beta2chi-nAChR than for alpha4beta2 nAChR. These studies are consistent with unexpected roles of nested C2 sequences in nAChR function. PMID- 15833892 TI - In situ naphthalene bioactivation and nasal airflow cause region-specific injury patterns in the nasal mucosa of rats exposed to naphthalene by inhalation. AB - Despite the fact that naphthalene (NA), a volatile, ubiquitous air pollutant, was recently identified as a probable human carcinogen, little is known about nasal cytotoxicity from inhaled NA. To define and compare acute nasal injury from inhalation and systemic NA exposures, male Sprague-Dawley rats were exposed to filtered air; 3.4 or 23.8 ppm NA by inhalation for 4 h; or to 0, 25, 50, 100, or 200 mg/kg NA via intraperitoneal injection. Severe cellular injury occurred exclusively in the olfactory mucosa 24 h postinhalation exposure to 3.4 ppm NA for 4 h. This level is significantly below both the current Occupational Safety and Health Administration standard (10 ppm; 8 h) for NA and the lowest observed adverse effect level (10 ppm; 2 years) for the incidence of rat olfactory neoplasms. Injury within the olfactory mucosa from inhaled NA was confined to the medial meatus, whereas systemic NA generated severe injury throughout the olfactory region. The pattern of nasal injury from inhaled NA in this study is consistent with previous studies of nasal airflow simulation within the olfactory region. The nonolfactory mucosa on the nasal septum, a high airflow region, metabolized naphthalene slowly, whereas the olfactory regions of the nasal septum and ethmoturbinates metabolized this substrate at high rates. This study concludes that 1) the incidence of acute nasal injury from systemic and inhaled NA correlates with the rates of regional microsomal NA metabolism and that 2) the nasal airflow pattern determines the pattern of olfactory mucosal injury from inhaled NA. PMID- 15833893 TI - Chemoresistance to depsipeptide FK228 [(E)-(1S,4S,10S,21R)-7-[(Z)-ethylidene] 4,21-diisopropyl-2-oxa-12,13-dithia-5,8,20,23-tetraazabicyclo[8,7,6]-tricos-16 ene-3,6,9,22-pentanone] is mediated by reversible MDR1 induction in human cancer cell lines. AB - Histone acetylation status, an epigenetic determinant of gene transcription, is controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). The potent HDAC inhibitor FK228 [(E)-(1S,4S,10S,21R)-7-[(Z)-ethylidene]-4,21 diisopropyl-2-oxa-12,13-dithia-5,8,20,23-tetraazabicyclo[8,7,6]-tricos-16-ene 3,6,9,22-pentanone] is a substrate for multidrug resistance protein (MDR1) and multidrug resistance-associated protein 1 (MRP1), both of which mediate FK228 resistance. To determine the mechanisms underlying acquired FK228 resistance, we developed four FK228-resistant cell lines from HCT-15, IGROV1, MCF7, and K562 cells by stepwise increases in FK228 exposure. Parent and resistant cells were characterized using a 70-oligomer cDNA microarray, real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and cytotoxicity assays. At both mRNA and protein levels, MDR1, but not MRP1 or other potential resistance genes, was strongly up-regulated in all resistant cell lines. HAT or HDAC activities were unaffected in resistant cells, consistent with a lack of cross-resistance to HDAC inhibitors that are not MDR1 substrates. FK228 was found to reversibly induce MDR1 expression by HDAC inhibition and subsequent histone hyperacetylation at the MDR1 promoter, as shown by real-time RT-PCR, Western blot, and chromatin immunoprecipitation. This study reveals a significant role of histone acetylation in MDR1 transcription, which seems to mediate FK228 resistance. PMID- 15833894 TI - Therapeutic actions of an insulin receptor activator and a novel peroxisome proliferator-activated receptor gamma agonist in the spontaneously hypertensive obese rat model of metabolic syndrome X. AB - Insulin resistance clusters with hyperlipidemia, impaired glucose tolerance, and hypertension as metabolic syndrome X. We tested a low molecular weight insulin receptor activator, demethylasterriquinone B-1 (DMAQ-B1), and a novel indole peroxisome proliferator-activated receptor gamma agonist, 2-(2-(4-phenoxy-2 propylphenoxy)ethyl)indole-5-acetic acid (PPEIA), in spontaneously hypertensive obese rats (SHROB), a genetic model of syndrome X. Agents were given orally for 19 days. SHROB showed fasting normoglycemia but impaired glucose tolerance after an oral load, as shown by increased glucose area under the curve (AUC) [20,700 mg x min/ml versus 8100 in lean spontaneously hypertensive rats (SHR)]. Insulin resistance was indicated by 20-fold excess fasting insulin and increased insulin AUC (6300 ng x min/ml versus 990 in SHR). DMAQ-B1 did not affect glucose tolerance (glucose AUC = 21,300) but reduced fasting insulin 2-fold and insulin AUC (insulin AUC = 4300). PPEIA normalized glucose tolerance (glucose AUC = 9100) and reduced insulin AUC (to 3180) without affecting fasting insulin. PPEIA also increased food intake, fat mass, and body weight gain (81 +/- 12 versus 45 +/- 8 g in untreated controls), whereas DMAQ-B1 had no effect on body weight but reduced subscapular fat mass. PPEIA but not DMAQ-B1 reduced blood pressure. In skeletal muscle, insulin-stimulated phosphorylation of the insulin receptor and insulin receptor substrate protein 1-associated phosphatidylinositol 3-kinase activity were decreased by 40 to 55% in SHROB relative to lean SHR. PPEIA, but not DMAQ-B1, enhanced both insulin actions. SHROB also showed severe hypertriglyceridemia (355 +/- 42 mg/dl versus 65 +/- 3 in SHR) attenuated by both agents (DMAQ-B1, 228 +/- 18; PPEIA, 79 +/- 3). Both these novel antidiabetic agents attenuate insulin resistance and hypertriglyceridemia associated with metabolic syndrome but via distinct mechanisms. PMID- 15833895 TI - Can claims, misleading information, and manufacturing issues regarding dietary supplements be improved in the United States? AB - The safety and effectiveness of over-the-counter (OTC) drugs are assessed through the Food and Drug Administration's (FDA's) OTC drug review. Prescription drugs are approved through the rigorous new drug application (NDA) process. In contrast, dietary supplements are regulated as foods, and the FDA must determine that a dietary supplement ingredient poses a "significant or unreasonable risk of illness or injury" instead of requiring the manufacturer to provide safety data. According to the FDA, there are more than 29,000 different dietary supplements available to consumers today. This momentum has its roots in consumer interest in health and self-care and suggests that Americans are searching for alternatives to conventional foods for physical and mental well being. The Committee on the Framework for Evaluating the Safety of Dietary Supplements was formed under the auspices of the Food and Nutrition Board that produced a report entitled Dietary Supplements: A Framework for Evaluating Safety. Categories of specific information identified for use are 1) human data, 2) animal studies, 3) in vitro experiments, and 4) information on related substances. Several factors were identified to guide the FDA in applying the framework. Two of these factors are expressed as follows: 1) "the appropriate scientific standard to be used to overturn this basic assumption of safety is to demonstrate significant or unreasonable risk, not prove that an ingredient is unsafe"; and 2) "approaches taken by diverse organizations and governmental bodies, both within and outside the United States, which evaluate the safety and at times efficacy of dietary supplement ingredients, vary in their relevance to the protection of the American public from risks associated with consumption of dietary supplement ingredients". PMID- 15833896 TI - Acute inhibition of cardiac monoamine oxidase A after tobacco smoke inhalation: validation study of [11C]befloxatone in rats followed by a positron emission tomography application in baboons. AB - The in vivo characteristics of [11C]befloxatone were assessed in myocardium of rats and monkeys. A complete multicompartmental model was developed to quantify monkey cardiac monoamine oxidase A (MAO-A) binding sites using positron emission tomography (PET) and was applied to assess the acute effects of inhalation of tobacco smoke. Unknown compounds contained in tobacco smoke inhibit brain MAO. In vitro, befloxatone inhibits selectively, competitively, and reversibly MAO-A in human tissues. [11C]Befloxatone (1.85 MBq) was i.v. injected into rats. Animals were sacrificed, dissected, and samples were assessed for radioactivity. Another group of rats was pretreated with clorgyline (10 mg/kg i.v.). Monkeys were injected with [11C]befloxatone (222-370 MBq), and the chest was imaged with PET for 2 h. Presaturation and displacement experiments were performed using unlabeled befloxatone. For quantification of myocardial binding sites (Bmax), [11C]befloxatone was first injected as a tracer dose (2.7-9.3 nmol) and 20 min later injected as a mixture of labeled and unlabeled befloxatone (labeled, 10.3 41.9 nmol; unlabeled, 407-765 nmol). In rodents, cardiac uptake was high (3.39 +/ 0.5% injected dose/g tissue) and strongly inhibited (80%) by clorgyline. In monkeys, administration of unlabeled befloxatone displaced 85% of cardiac radioactivity. Bmax was found to be 208 +/- 13 pmol ml(-1) tissue. Inhalation of tobacco smoke decreased Bmax: 150 +/- 6.2 pmol ml(-1), whereas nicotine did not. [11C]Befloxatone allows a good visualization of the heart. Cardiac MAO-A Bmax was quantified and a clear effect of acute inhalation of tobacco smoke was evidenced. Therefore, a single cigarette can interfere with the cardiac turnover of catecholamines. PMID- 15833897 TI - Dopamine agonist-induced yawning in rats: a dopamine D3 receptor-mediated behavior. AB - A specific role for the dopamine D3 receptor in behavior has yet to be elucidated. We now report that dopamine D2/D3 agonists elicit dose-dependent yawning behavior in rats, resulting in an inverted U-shaped dose-response curve. A series of experiments was directed toward the hypothesis that the induction of yawning is a D3 receptor-mediated effect, whereas the inhibition of the yawning observed at higher doses is due to competing D2 receptor activity. We compared several dopaminergic agonists with a range of in vitro D3 selectivity, including PD-128,907 [(S)-(+)-(4aR, 10bR)-3,4,4a,10b-tetrahydro-4-propyl-2H,5H [1]benzopyrano-[4,3-b]-1,4-oxazin-9-ol HCl], PD-128,908 [(R)-(-)-(4aS,10bS) 3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1]benzopyrano-[4,3-b]-1,4-oxazin-9-ol HCl], quinelorane [(5aR-trans)-5,5a,6,7,8, 9,9a,10-octahydro-6-propylpyrido[2,3 g]quinazolin-2-amine dihydrochloride], pramipexole (N'-propyl-4,5,6,7 tetrahydrobenzothiazole-2,6-diamine), 7-OH-DPAT [(+/-)-7-hydroxy-2 dipropylaminotetralin HBr], quinpirole [trans-(-)-(4aR)-4,4a,5,6,7,8, 8a,9 octahydro-5-propyl-1H-pyrazolo[3,4-g]quinoline HCl], bromocriptine [(+)-2-bromo 12'-hydroxy-2'-(1-methylethyl)-5'-(2-methylpropyl) ergotaman-3',6'-18-trione methanesulfonate], and apomorphine [(R)-(-)-5,6,6a,7-tetrahydro-6-methyl-4H dibenzo-[de,g]quinoline-10,11-diol HCl] with respect to their ability to induce yawning in rats. A series of D2/D3 antagonists differing in selectivity for D3 over D2 receptors were evaluated for their ability to alter the effects of the dopamine agonists. The antagonists L-741,626 (3-[4-(4-chlorophenyl)-4 hydroxypiperidin-l-yl]methyl-1H-indole), haloperidol (4-[4-(4-chlorophenyl)-4 hydroxy-1-piperidinyl]-1-(4-fluorophenyl)-1-butanone HCl), nafadotride (N-[(1 butyl-2-pyrrolidinyl)methyl]-4-cyano-1-methoxy-2-naphtha-lenecarboxamide), U99194 (2,3-dihydro-5,6-dimethoxy-N,N-dipropyl-1H-inden-2-amine maleate), SB-277011A (trans-N-[4-[2-(6-cyano-1,2,3,4-tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-4 quinolinecarboxamide), and PG01037 (N-{4-[4-(2,3-dichlorophenyl)-piperazin-1-yl] trans-but-2-enyl}-4-pyridine-2-yl-benzamide HCl) were used to determine effects on dose-response curves for D2/D3 agonist-induced yawning. In addition, the potential contribution of cholinergic and/or serotonergic mechanisms to the yawning response was investigated using a series of pharmacological tools including scopolamine [(a,S)-a-(hydroxymethyl)benzeneacetic acid (1a,2b,4b,5a,7b) 9-methyl-3-oxa-9-azatricyclo[3.3.1.02,4]-non7-yl ester hydrobromide], mianserin (1,2,3,4,10,14b-hexahydro-2-methyldibenzo[c,f]pyrazino[1,2-a]azepine HCl), and the D3-preferring antagonists nafadotride, U99194, SB-277011A, and PG01037 to differentially modulate yawning induced by PD-128,907, physostigmine [(3aS)-cis 1,2,3,3a,8,8a-hexahydro-1,3a,8-trimethylpyrrolo[2,3-b]indol-5-ol methylcarbamate hemisulfate], and N-[3-(trifluoromethyl)phenyl]piperazine HCl. The results of these experiments provide convergent evidence that dopamine D2/D3 agonist-induced yawning is a D3 agonist-mediated behavior, with subsequent inhibition of yawning being driven by competing D2 agonist activity. Thus, dopamine agonist-induced yawning may represent an in vivo method for selectively identifying D3 and D2 receptor-mediated activities. PMID- 15833898 TI - The ratio of constitutive androstane receptor to pregnane X receptor determines the activity of guggulsterone against the Cyp2b10 promoter. AB - Guggulsterone is the active ingredient in gugulipid, an organic extract of the Commiphora mukul plant. Gugulipid has been used for nearly 3000 years in Ayurvedic medicine, mainly as a treatment for arthritis. Herbal practitioners currently use gugulipid therapy in conditions as diverse as rheumatism, coronary artery disease, arthritis, hyperlipidemia, acne, and obesity. The active ingredient in gugulipid is guggulsterone, a plant sterol compound recently identified as a pregnane X receptor (PXR; NR1I2) ligand. We show herein that guggulsterone treatment represses the expression of cytochrome P450 2b10 (Cyp2b10) gene expression by inhibiting constitutive androstane receptor (CAR; NR1I3) activity in hepatocytes lacking functional PXR (PXR-knockout). We also show that PXR-CAR cross-talk determines the net activity of guggulsterone treatment toward Cyp2b10 gene expression. Using mammalian two-hybrid assays, we show that treatment with guggulsterone differentially affects protein cofactor recruitment to these two nuclear receptors. These data identify guggulsterone as an inverse agonist of the nuclear receptor CAR. When viewed together with the data showing that PXR and CAR expression is highly variable in different ethnic populations and that CAR expression is under the control of a circadian rhythm, our data provide important insight into the molecular mechanism of interindividual variability of drug metabolism. These data, together with the recent resolution of the crystal structures of PXR and CAR, will likely aid in the rational design of more specific CAR inverse agonists that are currently viewed as potential antiobesity drugs. PMID- 15833899 TI - Suramin promotes proliferation and scattering of renal epithelial cells. AB - Primary cultures of renal proximal tubules are known to recapitulate several early events in the process of renal regeneration following injury. In this study, we show that suramin, a polysulfonated naphthylurea, stimulates outgrowth, scattering, and proliferation of primary cultures of renal proximal tubule cells (RPTC). These responses were comparable to those produced by epidermal growth factor (EGF). However, AG-1478 [4-(3'-chloroanilino)-6,7-dimethoxy-quinazoline], a specific inhibitor of the EGF receptor, blocked EGF but not suramin-induced RPTC outgrowth, scattering, and proliferation. Suramin stimulated phosphorylation of Akt, a downstream kinase of phosphoinositide 3-kinase (PI3K), extracellular signaling-regulated kinase 1/2 (ERK1/2), and Src, but not the EGF receptor. Blockade of Src, but not the EGF receptor, inhibited Akt and ERK1/2 phosphorylation. Furthermore, inactivation of PI3K with LY294002 [2 (4morpholinyl)-8-phenyl-4H-1-benzopyran-4-one] blocked suramin-induced RPTC outgrowth, scattering, and proliferation, whereas blockade of ERK1/2 had no effect. These data identify novel effects of suramin in RPTC outgrowth, scattering, and proliferation. Furthermore, suramin-induced outgrowth, scattering, and proliferation of RPTC are through Src-mediated activation of the PI3K pathway but not ERK1/2 or the EGF receptor. PMID- 15833900 TI - Estimating human drug oral absorption kinetics from Caco-2 permeability using an absorption-disposition model: model development and evaluation and derivation of analytical solutions for k(a) and F(a). AB - Intestinal transcellular permeability (P(m)), measured across cell lines such as Caco-2 cells in vitro, is often used for assessing oral drug absorption potential in humans. However, the quantitative link between in vitro permeability and apparent in vivo absorption kinetics, based on drug appearance in plasma, is poorly understood. In the current study, a novel absorption-disposition kinetic model that links traditional pharmacokinetic and mass transfer models was developed. Analytical solutions of k(a) and F(a) were deduced, and using Caco-2 permeability, F(a) in humans was predicted for 51 structurally diverse compounds. Predicted F(a) values were similar to and correlated highly with their corresponding experimental values with an average error of 1.88 +/- 1.06% (-17 to 22%) and r2 = 0.934. Simulated concentration profiles for 17 of 18 drugs corresponded to observed plasma concentration profiles in healthy volunteers. The equilibrium solution for k(a) (k(a,eq)) was found to be a key determinant of F(a), whereas under sink conditions, k(a) is likely to be a determinant of plasma concentration kinetics. The current version of the model offers a quantitative approach for predicting human oral absorption kinetics from in vitro permeability. It also establishes, for the first time, a quantitative link between P(m) and k(a) and between k(a,eq) and F(a). This will facilitate better in vitro or in situ-in vivo correlations since it establishes a basis for incorporating permeability coefficients from the various experimental formats based on drug loss or appearance that are commonly used in the laboratory for permeability determination. PMID- 15833901 TI - Maintenance and decay of past behavior influences: anchoring attitudes on beliefs following inconsistent actions. AB - Three studies investigated the influence of past behavior on the stability of the attitudes it elicits. In Experiment 1, the effect of a bogus behavior feedback was long lasting when people engaged in biased scanning, presumably because this process elicits behavior-consistent beliefs. In contrast, the effect of the feedback decayed when participants were forced to consider whether the behavior might have undesirable outcomes. A second experiment using a different behavioral paradigm and a field study further supported the interpretation that individuals resolve conflict between a past behavior and subsequent beliefs about it by aligning attitudes with beliefs instead of behavior. PMID- 15833902 TI - Emotional disclosure and closeness toward offenders. AB - Two studies tested whether emotional disclosure increases feelings of closeness toward offenders. In Study 1, participants recalled either someone who had offended them or a neutral acquaintance. "Disclosure" participants then expressed their thoughts and feelings regarding their targets, and "suppression" participants described their targets nonemotionally. As predicted, disclosure increased closeness toward offenders but not toward acquaintances. Study 2 extended these results by including a good friend to test whether disclosure selectively increases closeness toward offenders, and not simply toward any person who evokes strong feelings. This prediction was confirmed. Furthermore, the disclosure effect remained reliable even after controlling for mood. Studies 1 and 2 also showed that closeness toward offenders, but not toward friends or acquaintances, was positively related to the proportion of emotion-related words disclosed. Collectively, these findings suggest that confronting the emotions associated with an offense may be an important first step toward forgiveness. PMID- 15833903 TI - Is short-term mating the maladaptive result of insecure attachment? A test of competing evolutionary perspectives. AB - Different evolutionary perspectives offer competing views on short-term mating and attachment. Some theories argue that short-term mating results from insecure attachment, particularly the maladaptive attachment features of low self-esteem, interpersonal distrust, social avoidance, and emotional instability. Other theories posit that short-term mating is adaptive, having evolved as an ecologically contingent reproductive strategy. In this view, short-term mating has multiple origins-developmental, heritable, and situational-and may not be associated with the maladaptive traits of insecure attachment. Across several different cultures, short-term mating was moderately associated with insecure attachment, and insecure attachment was strongly related to maladaptive personality. However, short-term mating was largely independent of maladaptive personality. In some cases, especially among young men, short-term mating was associated with adaptive personality traits, especially higher self-esteem. It is argued that although insecure romantic attachment is somewhat associated with short-term mating, the causal links between early parent-child attachment and short-term mating are rather limited. PMID- 15833904 TI - Perceivers' responses to in-group and out-group members who blame a negative outcome on discrimination. AB - The authors extend recent research concerning the social costs of claiming discrimination by examining men's and women's responses to in-group and out-group targets who either blamed a failing grade on discrimination or answer quality. Although participants generally responded more negatively to targets who blamed discrimination, rather than answer quality, dislike was greatest and gender group identification was lowest when participants evaluated an in-group target. Moreover, an in-group target who claimed discrimination was perceived as avoiding personal responsibility for outcomes to a greater extent than was a similar out group target. Perceptions that the target avoided outcome responsibility by claiming discrimination were shown to mediate the relationship between attribution type and dislike of the in-group target. The authors discuss their results in terms of intragroup processes and suggest that social costs may especially accrue for in-group members when claiming discrimination has implications for the in-group's social identity. PMID- 15833905 TI - Putting stereotype content in context: image theory and interethnic stereotypes. AB - Two studies apply intergroup image theory to better understand divergent interethnic images and to highlight the important role of intergroup context and perceived intergroup relations in shaping the content of social stereotypes. Image theory hypothesizes that specific interethnic stereotypes arise from specific patterns of perceived intergroup competition, relative power, and relative cultural status. Results from surveying Black, White, and Native Americans' appraisals of intergroup relations and reported outgroup stereotypes in various intergroup contexts suggest that the content of outgroup stereotypes varies systematically as a function of the perceived state of intergroup relations and the intergroup context in which these groups are situated. The data reported from both studies establish the importance of examining social stereotypes from a functional perspective in the context of intergroup relations. PMID- 15833906 TI - Connecting the horizontal dimension of social comparison with self-worth and self confidence. AB - Multilevel modeling of undergraduates (N = 229) event-contingent records of naturalistic social comparison experiences revealed distinct correlates of the horizontal (similar-different) dimension and vertical (better-worse) dimension of comparisons. Complementing past studies showing associations between the horizontal dimension and communal dispositions and experiences, the current study showed that the horizontal dimension also is associated with agentic dispositions and experiences such as self-worth and self-confidence. For example, participants perceived more similarity when comparing with targets' desirable attributes than with targets' undesirable attributes and perceiving similarities with desirable target attributes (and dissimilarities with undesirable target attributes) enhanced their self-confidence. Participants higher in self-worth (high in self esteem and low in depression) were more discriminating in their experiences of similarity and connection; specifically, they reported more similarity and connection when targets' attributes were desirable but less connection the more targets' attributes were inferior to their own. PMID- 15833907 TI - Uncertainty reduction, self-enhancement, and ingroup identification. AB - Two experiments tested the prediction that uncertainty reduction and self enhancement motivations have an interactive effect on ingroup identification. In Experiment 1 (N = 64), uncertainty and group status were manipulated, and the effect on ingroup identification was measured. As predicted, low-uncertainty participants identified more strongly with a high- than low-status group, whereas high-uncertainty participants showed no preference; and low-status group members identified more strongly under high than low uncertainty, whereas high-status group members showed no preference. Experiment 2 (N = 210) replicated Experiment 1, but with a third independent variable that manipulated how prototypical participants were of their group. As predicted, the effects obtained in Experiment 1 only emerged where participants were highly prototypical. Low prototypicality depressed identification with a low-status group under high uncertainty. The implications of these results for intergroup relations and the role of prototypicality in social identity processes are discussed. PMID- 15833908 TI - Written emotional expression and emotional well-being: the moderating role of fear of rejection. AB - Empirical research shows that individuals high in fear of rejection typically report low levels of perceived social support and are more vulnerable to stressful experiences. At the same time, writing about stressful experiences in an emotional way seems to help people adapt to current stressors and not-yet assimilated stressful experiences. Therefore, the authors suggest that written emotional expression may be a particularly effective strategy to manage negative emotions for individuals high in fear of rejection. Three studies were conducted to test these assumptions. Study 1 found that high fear of rejection is linked to a lack of perceived social support. Longitudinal Studies 2 and 3 supported our main hypothesis, demonstrating that written emotional expression is linked to lower levels of negative mood among individuals high (but not among individuals low) in fear of rejection. PMID- 15833909 TI - Do unto others as they do unto you: reciprocity and social identification as determinants of ingroup favoritism. AB - Predictions from the bounded and unbounded reciprocity hypotheses and from social identity theory (SIT) were examined in a minimal group experiment in which ingroup outcome dependence, outgroup outcome dependence, and the strength of social identity were orthogonally manipulated. Both ingroup and outgroup outcome dependence affected reward allocations. Participants made more ingroup-favoring reward allocations across all conditions. The identification manipulation produced hypothesized effects on social identification measures and marginal effects of identification on reward allocations in the no-dependence condition. Support was found for both an unbounded and bounded version of the reciprocity hypothesis and marginal support for a SIT approach to intergroup discrimination. The study highlights insufficiencies of both theoretical approaches and suggests possibilities for integration and elaboration. PMID- 15833910 TI - Stereotyping, self-affirmation, and the cerebral hemispheres. AB - The authors used the processing characteristics of the left and right cerebral hemispheres to gain some insight into the relation between self-affirmation and stereotyping. In Study 1, self-affirmation led to greater stereotyping (of a librarian) and to greater left hemisphere activation, which in turn mediated the self-affirmation/stereotyping relationship. Study 2 replicated these results but also found that self-affirmation decreased stereotyping for a stigmatized target. However, relative hemisphere activation did not mediate this self affirmation/stereotyping relationship. These studies showed that self-affirmation can either increase or decrease stereotyping depending on the status of the target and that relative hemisphere activation may provide clues as to underlying processes of stereotyping. In both studies, relative hemisphere activation was assessed using a line bisection task. Discussion focuses on possible mechanisms of different kinds of stereotyping and on the ways in which a consideration of relative hemisphere activation could help researchers gain insights into those mechanisms. PMID- 15833911 TI - Unique coexpression in osteoblasts of broadly expressed genes accounts for the spatial restriction of ECM mineralization to bone. AB - Extracellular matrix (ECM) mineralization is a physiological process in bone and a pathological one in soft tissues. The mechanisms determining the spatial restriction of ECM mineralization to bone physiologically are poorly understood. Here we show that a normal extracellular phosphate concentration is required for bone mineralization, while lowering this concentration prevents mineralization of any ECM. However, simply raising extracellular phosphate concentration is not sufficient to induce pathological mineralization, this is because of the presence in all ECMs of pyrophosphate, an inhibitor of mineralization. ECM mineralization occurs only in bone because of the exclusive coexpression in osteoblasts of Type I collagen and Tnap, an enzyme that cleaves pyrophosphate. This dual requirement explains why Tnap ectopic expression in cells producing fibrillar collagen is sufficient to induce pathological mineralization. This study reveals that coexpression in osteoblasts of otherwise broadly expressed genes is necessary and sufficient to induce bone mineralization and provides evidence that pathological mineralization can be prevented by modulating extracellular phosphate concentration. PMID- 15833912 TI - Pervasive regulation of Drosophila Notch target genes by GY-box-, Brd-box-, and K box-class microRNAs. AB - Although hundreds of distinct animal microRNAs (miRNAs) are known, the specific biological functions of only a handful are understood at present. Here, we demonstrate that three different families of Drosophila miRNAs directly regulate two large families of Notch target genes, including basic helix-loop-helix (bHLH) repressor and Bearded family genes. These miRNAs regulate Notch target gene activity via GY-box (GUCUUCC), Brd-box (AGCUUUA), and K-box (cUGUGAUa) motifs. These are conserved sites in target 3'-untranslated regions (3'-UTRs) that are complementary to the 5'-ends of miRNAs, or "seed" regions. Collectively, these motifs represent >40 miRNA-binding sites in Notch target genes, and we show all three classes of motif to be necessary and sufficient for miRNA-mediated regulation in vivo. Importantly, many of the validated miRNA-binding sites have limited pairing to miRNAs outside of the "box:seed" region. Consistent with this, we find that seed-related miRNAs that are otherwise quite divergent can regulate the same target sequences. Finally, we demonstrate that ectopic expression of several Notch-regulating miRNAs induces mutant phenotypes that are characteristic of Notch pathway loss of function, including loss of wing margin, thickened wing veins, increased bristle density, and tufted bristles. Collectively, these data establish insights into miRNA target recognition and demonstrate that the Notch signaling pathway is a major target of miRNA-mediated regulation in Drosophila. PMID- 15833914 TI - Wnt signaling in the intestinal epithelium: from endoderm to cancer. AB - The Wnt pathway controls cell fate during embryonic development. It also persists as a key regulator of homeostasis in adult self-renewing tissues. In these tissues, mutational deregulation of the Wnt cascade is closely associated with malignant transformation. The intestinal epithelium represents the best understood example for the closely linked roles of Wnt signaling in homeostatic self-renewal and malignant transformation. In this review, we outline current understanding of the physiological role of Wnt signaling in intestinal biology. From this perspective, we then describe how mutational subversion of the Wnt cascade leads to colorectal cancer. PMID- 15833913 TI - Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint. AB - The ATR-dependent DNA damage response pathway can respond to a diverse group of lesions as well as inhibitors of DNA replication. Using the Xenopus egg extract system, we show that lesions induced by UV irradiation and cis-platinum cause the functional uncoupling of MCM helicase and DNA polymerase activities, an event previously shown for aphidicolin. Inhibition of uncoupling during elongation with inhibitors of MCM7 or Cdc45, a putative helicase cofactor, results in abrogation of Chk1 phosphorylation, indicating that uncoupling is necessary for activation of the checkpoint. However, uncoupling is not sufficient for checkpoint activation, and DNA synthesis by Polalpha is also required. Finally, using plasmids of varying size, we demonstrate that all of the unwound DNA generated at a stalled replication fork can contribute to the level of Chk1 phosphorylation, suggesting that uncoupling amplifies checkpoint signaling at each individual replication fork. Taken together, these observations indicate that functional uncoupling of MCM helicase and DNA polymerase activities occurs in response to multiple forms of DNA damage and that there is a general mechanism for generation of the checkpoint-activating signal following DNA damage. PMID- 15833915 TI - Metal-responsive transcription factor (MTF-1) handles both extremes, copper load and copper starvation, by activating different genes. AB - From insects to mammals, metallothionein genes are induced in response to heavy metal load by the transcription factor MTF-1, which binds to short DNA sequence motifs, termed metal response elements (MREs). Here we describe a novel and seemingly paradoxical role for MTF-1 in Drosophila in that it also mediates transcriptional activation of Ctr1B, a copper importer, upon copper depletion. Activation depends on the same type of MRE motifs in the upstream region of the Ctr1B gene as are normally required for metal induction. Thus, a single transcription factor, MTF-1, plays a direct role in both copper detoxification and acquisition by inducing the expression of metallothioneins and of a copper importer, respectively. PMID- 15833916 TI - Medial floor plate formation in zebrafish consists of two phases and requires trunk-derived Midkine-a. AB - The medial floor plate (MFP) organizes the specification of neurons and outgrowth of axons in the ventral spinal cord of vertebrates. We show that the growth factor Midkine-a, expressed in the paraxial mesoderm, is required for formation of the MFP in zebrafish. Our epistatic analyses demonstrate that development of MFP comprises two independent sequential phases. Following initial MFP induction in the gastrula organizer, Midkine-a regulates allocation of MFP cells during subsequent development. Thus in zebrafish, trunk-derived signals are required for complete MFP formation from a common pool of organizer-derived midline precursor cells. PMID- 15833917 TI - Genome-wide identification of Isw2 chromatin-remodeling targets by localization of a catalytically inactive mutant. AB - Isw2 ATP-dependent chromatin-remodeling activity is targeted to early meiotic and MATa-specific gene promoters in Saccharomyces cerevisiae. Unexpectedly, preferential cross-linking of wild-type Isw2p was not detected at these loci. Instead, the catalytically inactive Isw2p-K215R mutant is enriched at Isw2 targets, suggesting that Isw2p-K215R, but not wild-type Isw2p, is a sensitive chromatin immunoprecipitation (ChIP) reagent for marking sites of Isw2 activity in vivo. Genome-wide ChIP analyses confirmed this conclusion and identified tRNA genes (tDNAs) as a new class of Isw2 targets. Loss of Isw2p disrupted the periodic pattern of Ty1 integration upstream of tDNAs, but did not affect transcription of tDNAs or the associated Ty1 retrotransposons. In addition to identifying new Isw2 targets, our localization studies have important implications for the mechanism of Isw2 association with chromatin in vivo. Target specific enrichment of Isw2p-K215R, not wild-type Isw2p, suggests that Isw2 is recruited transiently to remodel chromatin structure at these sites. In contrast, we found no evidence for Isw2 function at sites preferentially enriched by wild type Isw2p, leading to our proposal that wild-type Isw2p cross-linking reveals a scanning mode of the complex as it surveys the genome for its targets. PMID- 15833918 TI - TFIIIB subunit Bdp1p is required for periodic integration of the Ty1 retrotransposon and targeting of Isw2p to S. cerevisiae tDNAs. AB - Retrotransposons are RNA elements that reverse transcribe their RNA genomes and make a cDNA copy that is inserted back into a new genomic location by the element encoded integrase protein. Ty1 is a long terminal repeat (LTR) retrotransposon in Saccharomyces cerevisiae that inserts into an approximately 700-bp integration window upstream of tRNA genes with a periodicity of approximately 80 bp. ATP dependent chromatin remodeling by Isw2 upstream of tRNA genes leads to changes in chromatin structure and Ty1 integration site selection. We show that the N terminus of Bdp1p, a component of the RNA polymerase III transcription factor TFIIIB, is required for periodic integration of Ty1 into the integration window. Deletion of the Bdp1p N terminus and mutation of ISW2 result in similar disruption of nucleosome positioning upstream of some tRNA genes, and the N terminal domain of Bdp1p is required for targeting of Isw2 complex to tRNA genes. This study provides the first example for recruitment of an ATP-dependent chromatin-remodeling factor by a general transcription factor in vivo. PMID- 15833919 TI - Delayed, asynchronous, and reversible T-lineage specification induced by Notch/Delta signaling. AB - Using the OP9-DL1 system to deliver temporally controlled Notch/Delta signaling, we show that pluripotent hematolymphoid progenitors undergo T-lineage specification and B-lineage inhibition in response to Notch signaling in a delayed and asynchronous way. Highly enriched progenitors from fetal liver require > or =3 d to begin B- or T-lineage differentiation. Clonal switch-culture analysis shows that progeny of some single cells can still generate both B- and T lineage cells, after 1 wk of continuous delivery or deprivation of Notch/Delta signaling. Notch signaling induces T-cell genes and represses B-cell genes, but kinetics of activation of lineage-specific transcription factors are significantly delayed after induction of Notch target genes and can be temporally uncoupled from the Notch response. In the cells that initiate T-cell differentiation and gene expression most slowly in response to Notch/Delta signaling, Notch target genes are induced to the same level as in the cells that respond most rapidly. Early lineage-specific gene expression is also rapidly reversible in switch cultures. Thus, while necessary to induce and sustain T-cell development, Notch/Delta signaling is not sufficient for T-lineage specification and commitment, but instead can be permissive for the maintenance and proliferation of uncommitted progenitors that are omitted in binary-choice models. PMID- 15833920 TI - WUSCHEL is a primary target for transcriptional regulation by SPLAYED in dynamic control of stem cell fate in Arabidopsis. AB - SNF2 chromatin-remodeling ATPases play an important role in ensuring proper development in higher eukaryotes by controlling accessibility of cis-regulatory DNA regions to transcription factors and to the transcriptional machinery. However, the biological targets controlled by these ATPases are largely unknown. Using genetic and molecular analyses we have identified WUSCHEL (WUS) as a biologically important target of the SNF2-class ATPase SPLAYED (SYD) in the shoot apical meristem of Arabidopsis. We present evidence that SYD is recruited to the WUS promoter and that it is involved in regulation of the stem cell pool maintenance via direct transcriptional control of this master regulator. PMID- 15833921 TI - New restrictions on celecoxib (Celebrex) use and the withdrawal of valdecoxib (Bextra). PMID- 15833922 TI - Shining a light on sunscreens. PMID- 15833923 TI - Embryo culture conditions: what embryos like best. PMID- 15833924 TI - White adipose tissue, inert no more! PMID- 15833926 TI - Arsenite decreases CYP3A4 and RXRalpha in primary human hepatocytes. AB - Arsenic is a naturally occurring, worldwide contaminant implicated in numerous pathological conditions in humans, including cancer and several forms of liver disease. One of the contributing factors to these disorders may be the alteration of cytochrome P450 (P450) levels by arsenic. P450s are involved in the oxidative metabolism and elimination of numerous toxic chemicals. CYP3A4, a major P450 in humans, is involved in the metabolism of half of all currently used drugs. Acute exposure to arsenite decreases the induction of CYP1A1/2 proteins and activities in cultured human hepatocytes, as well as CYP3A23 in cultured rat hepatocytes. Here, in primary cultures of human hepatocytes, we assessed the effects of acute arsenite exposure on CYP3A4 and several transcription factors involved in CYP3A4 expression. The concentrations of arsenite used in these studies were nontoxic to the hepatocytes and failed to elicit an oxidative response. Treatment with arsenite in the presence of CYP3A4 inducers, rifampicin (Rif) or phenobarbital, caused major decreases in CYP3A4 mRNA, protein, and activity. In addition, the levels of CYP3A4 in untreated cells were decreased following arsenite treatment. Transcription of the CYP3A4 gene is primarily regulated by heterodimers of the retinoid X receptor alpha (RXRalpha) and the pregnane X receptor (PXR). We found that arsenite failed to affect expression of PXR or the transcription factor Sp1, yet caused a significant decrease in PXR responsiveness to Rif. Arsenite caused a large decrease in nuclear RXRalpha protein and, to a lesser extent, RXRalpha mRNA. These results suggest that arsenite inhibits both untreated and induced CYP3A4 transcription in primary human hepatocytes by decreasing the activity of PXR, as well as expression of the nuclear receptor RXRalpha. PMID- 15833927 TI - Induction of CYP1A in the beagle dog by an inhibitor of kinase insert domain containing receptor: differential effects in vitro and in vivo on mRNA and functional activity. AB - Compound I [3-[5-(4-methanesulfonyl-piperazin-1-ylmethyl)-1H-indol-2-yl]-1H quinolin-2-one] is a potent inhibitor of human kinase insert domain-containing receptor (KDR kinase), which is under investigation for the treatment of cancer. Bile duct-cannulated male beagle dogs were administered 6 mg/kg compound I q.d. for 14 days. There was an approximately 2.5-fold decrease in the mean plasma area under the curve of I on days 7 and 14 (approximately 11.3 microM . h), relative to day 1 (28.2 microM . h). In the dog, compound I was eliminated by metabolism, with a major pathway being aromatic hydroxylation and subsequent sulfation to form the metabolite M3. Metabolic profiling suggested that the pathway leading to the formation of the sulfated conjugate M3 was induced upon multiple dosing of I. Studies conducted in vitro suggested that CYP1A1/2 was responsible for the formation of the hydroxylated metabolite, which is sulfated to yield M3. Additional studies confirmed induction of CYP1A protein and activity in the livers of dogs treated with I. However, studies in a dog hepatocyte model of induction showed a surprising decrease both in CYP1A mRNA and enzymatic activity in the presence of I, emphasizing the need to consider the results from a variety of in vitro and in vivo studies in deriving an understanding of the metabolic fate of a drug candidate. It is concluded that the autoinduction observed after multiple treatments with compound I occurs since compound I is both an inducer and a substrate for dog CYP1A. PMID- 15833928 TI - CYP3A5 genetic polymorphisms in different ethnic populations. AB - Cyp3A5 activity varies within any given ethnic population, suggesting that genetic variation within the Cyp3A5 gene may be the most important contributor to interindividual and interracial differences in Cyp3A-dependent drug clearance and response. The full extent of Cyp3A5 polymorphism in a white and an indigenous African population was analyzed using DNA direct sequencing procedures. The presence of 10 and 12 single nucleotide polymorphisms was detected in the white and African samples, respectively. Thirteen novel mutations occurring at low frequencies were identified in these populations. Significant differences were observed in the distribution of Cyp3A5*3, Cyp3A5*6, and Cyp3A5*7 alleles among white and African populations. The frequency of Cyp3A5*3 allele in white Canadians (approximately 93%) is higher than in Zimbabweans (77.6%) (p < 0.001). In contrast, Cyp3A5*6 and Cyp3A5*7 alleles are relatively frequent in African subjects (10-22%) but absent in white subjects (p < 0.001). These differences may reflect evolutionary pressures generated by environmental factors in geographically distinct regions. However, the genetic polymorphism of Cyp3A5 alone does not explain the interindividual differences in Cyp3A-mediated metabolism. PMID- 15833929 TI - Induction of the multidrug resistance-associated protein family of transporters by chemical activators of receptor-mediated pathways in mouse liver. AB - The multidrug resistance-associated proteins (Mrp) are ATP-dependent transporters that export a variety of conjugated and unconjugated compounds out of cells. There are nine identified Mrp transporters in humans, with murine orthologs for all except Mrp8. Because nuclear receptors mediate induction of phase I enzymes, Mrp transporter expression might be similarly regulated by these receptors to coordinate metabolism and export of chemicals from liver. To test the hypothesis that Mrp expression may be coordinately regulated with phase I enzyme expression in liver, 15 different compounds were given representing known transcriptionally mediated pathways: aryl hydrocarbon receptor (AhR), pregnane X receptor (PXR), constitutive androstane receptor (CAR), peroxisome proliferator-activated receptor alpha (PPARalpha), and nuclear factor-E2-related factor 2 (Nrf2). Each of these compounds induced expression of their respective target enzyme in liver, demonstrating that the chemical regimens were effective. The AhR ligands [2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), polychlorinated biphenyl 126 (PCB126), and beta-naphthoflavone] induced Mrp2, -3, -5, and -6 mRNA expression. The CAR activator 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP) induced Mrp2, -3, 4, -6, and -7 mRNA expression. Mrp3 was also induced by two other CAR activators phenobarbital and diallyl sulfide, two PXR ligands, pregnenalone-16alpha carbonitrile and spironolactone, and the PPARalpha ligands clofibrate, ciprofibrate, and diethylhexylphthalate. The Nrf2 activators (butylated hydroxyanisole, oltipraz, and ethoxyquin) induced Mrp2-6. In conclusion, a variety of mechanisms are suggested for Mrp3 induction, including AhR, CAR, PXR, PPARalpha, and Nrf2, whereas on a whole, a predominant role for AhR and Nrf2 in hepatic induction of the Mrp family was observed. Thus, these specific transcription factors are implicated in regulation of both drug metabolism and efflux transport. PMID- 15833930 TI - Differential rates of glucuronidation for 7-ethyl-10-hydroxy-camptothecin (SN-38) lactone and carboxylate in human and rat microsomes and recombinant UDP glucuronosyltransferase isoforms. AB - 7-ethyl-10-hydroxy-camptothecin (SN-38), the active metabolite of the anti-cancer agent irinotecan, contains a lactone ring that equilibrates with a carboxylate form. Since SN-38 lactone is the active and toxic form, it is prudent to examine whether the more soluble carboxylate is a surrogate for SN-38 lactone conjugation. Therefore, relative rates of glucuronidation and isoform specificity of SN-38 lactone and carboxylate were characterized. The stability of SN-38 lactone and carboxylate in incubation mixtures of microsomes and UDP glucuronosyltransferase (UGT) isoforms was used to determine optimal incubation times. Microsomal incubations were conducted using rat and human intestinal and hepatic microsomes and human and rat recombinant UGT1A isoforms. Where estimates of lactone and carboxylate glucuronidation rates could not be established due to short incubation times and detection limits, kinetic modeling was used to recover these rate constants. The stability experiments revealed that the lactone was stabilized by rat microsomes, however, the opposite was observed in human microsomes and recombinant isoforms. For all tissues and most UGT isoforms examined, the lactone consistently had catalytic rates up to 6-fold greater than the carboxylate. The rank order of glucuronidation for both SN-38 lactone and carboxylate was 1A7 > 1A1 > 1A9 > 1A8 and 1A7 > 1A8 > 1A1 for human and rat isoforms, respectively. This study provides further support that SN-38 lactone and carboxylate may be considered pharmacokinetically distinct agents. The in vivo impact of this conjugation difference is unknown, since variations in protein binding and transport proteins may affect intracellular concentrations of the lactone or carboxylate. PMID- 15833931 TI - Disposition and metabolism of [2-14C]quercetin-4'-glucoside in rats. AB - Quercetin-4'-glucoside is a major flavonol in onions, and this study investigated the absorption and fate of radiolabeled quercetin-4'-glucoside in rats. Rats ingested [2-(14)C]quercetin-4'-glucoside and the distribution of radioactivity throughout the body was determined after 0.5, 1, 2, and 5 h. The gastrointestinal tract, liver, kidney, and plasma were extracted, and radiolabeled components were identified and quantified using high-performance liquid chromatography with on line radioactivity detection and tandem mass spectrometry. Two hours after dosing, all the [2-(14)C]quercetin-4'-glucoside had been metabolized. More than 85% of the ingested radioactivity was present in the gastrointestinal tract at all time points with approximately 6% being absorbed and present in blood and internal organs, primarily the liver and kidneys. More than 95% of the absorbed radioactivity was in the form of >20 different methylated glucuronated and/or sulfated quercetin conjugates. Five hours after ingestion, the main radiolabeled metabolites were quercetin diglucuronides in the gut, liver, and kidneys and glucuronyl sulfates of methylated quercetin in plasma. The main site of quercetin metabolism seemed to be the gastrointestinal tract. Quercetin metabolites may have a major influence on the gut mucosal epithelium and on colonic disease. PMID- 15833932 TI - The human obesity gene map: the 2004 update. AB - This paper presents the eleventh update of the human obesity gene map, which incorporates published results up to the end of October 2004. Evidence from single-gene mutation obesity cases, Mendelian disorders exhibiting obesity as a clinical feature, transgenic and knockout murine models relevant to obesity, quantitative trait loci (QTLs) from animal cross-breeding experiments, association studies with candidate genes, and linkages from genome scans is reviewed. As of October 2004, 173 human obesity cases due to single-gene mutations in 10 different genes have been reported, and 49 loci related to Mendelian syndromes relevant to human obesity have been mapped to a genomic region, and causal genes or strong candidates have been identified for most of these syndromes. There are 166 genes which, when mutated or expressed as transgenes in the mouse, result in phenotypes that affect body weight and adiposity. The number of QTLs reported from animal models currently reaches 221. The number of human obesity QTLs derived from genome scans continues to grow, and we have now 204 QTLs for obesity-related phenotypes from 50 genome-wide scans. A total of 38 genomic regions harbor QTLs replicated among two to four studies. The number of studies reporting associations between DNA sequence variation in specific genes and obesity phenotypes has also increased considerably with 358 findings of positive associations with 113 candidate genes. Among them, 18 genes are supported by at least five positive studies. The obesity gene map shows putative loci on all chromosomes except Y. Overall, >600 genes, markers, and chromosomal regions have been associated or linked with human obesity phenotypes. The electronic version of the map with links to useful publications and genomic and other relevant sites can be found at http://obesitygene.pbrc.edu. PMID- 15833933 TI - Gastric motor and sensory functions in obesity. AB - In the vast majority of affected individuals, obesity involves overconsumption of food relative to calorie requirements. The sensory function of the stomach may play a key role in the cessation of food ingestion. This sensation of the stomach is, in part, determined by its motor functions, such as tone and compliance and the rate of emptying. However, studies of gastric emptying in normal-weight and obese persons have shown inconsistent results. Gastric capacity was larger in obese persons when tested with an intragastric latex balloon filled with water. In contrast, other studies using the barostat or imaging (single-photon emission computed tomography) techniques reported no differences in gastric volume or compliance between obese and lean subjects. On the other hand, increased body mass and fasting gastric volume are independently associated with delayed satiation under standard laboratory conditions of food ingestion. These data suggest that changes in gastric motor and sensory functions in obesity may present useful targets to prevent and treat obesity. Further well-controlled, validated studies are needed to clarify the potential role of altering the stomach's function as a means of controlling food intake in obesity. PMID- 15833934 TI - Leptin receptor gene variation predicts weight change in subjects with impaired glucose tolerance. AB - The leptin receptor (OB-R) gene is a promising candidate gene for type 2 diabetes, because leptin and its receptor play an important role in insulin secretion and the development of obesity. Therefore, we studied whether the pentanucleotide insertion polymorphism of the 3'-untranslated region (3'UTR) of the OB-R gene has an influence on the conversion from impaired glucose tolerance (IGT) to type 2 diabetes in the STOP-Noninsulin-Dependent Diabetes Mellitus trial. The STOP trial was a longitudinal, double-blind, placebo-controlled randomized trial that included 1429 subjects with IGT from high-risk populations. Using the restriction fragment length polymorphism method, we genotyped 770 subjects whose DNA was available for the insertion/deletion polymorphism of the 3'UTR of the OB-R gene. We did not find a relationship between the OB-R polymorphism and the conversion from IGT to type 2 diabetes (p = 0.747). However, the insertion allele was associated with a significant reduction in weight (p = 0.016), BMI (p = 0.009), and waist circumference (p = 0.006) in all subjects. Women carrying the I allele had a larger waist circumference change (p = 0.036), whereas men lost more weight and had a greater decrease in BMI. The pentanucleotide insertion/deletion polymorphism in the 3'UTR of the OB-R gene did not influence the conversion to type 2 diabetes in obese patients with IGT. However, this polymorphism was associated with a significant weight change, suggesting that it may potentially modulate the risk for type 2 diabetes. PMID- 15833935 TI - Apolipoprotein A-II polymorphism and visceral adiposity in African-American and white women. AB - To determine the association between the -265 T to C substitution in the apolipoprotein A-II (APOA-II) gene and levels of visceral adipose tissue (VAT) in a group of premenopausal African-American and white women, we genotyped 237 women (115 African-American and 122 white) for this polymorphism. Body composition was assessed by DXA, and VAT was determined from a single computed tomography scan. In addition to VAT, we examined the association between the polymorphism and other phenotypes (total body fat, total abdominal adipose tissue, and subcutaneous abdominal adipose tissue). The mutant C allele in the APOA-II gene was less frequent in African-American compared with white women, 23% vs. 36%, respectively (p < 0.01). VAT was significantly higher in carriers of the C allele compared with noncarriers after adjustment for total body fat (p < 0.05). When separate analyses by ethnic group were conducted, the association between the polymorphism and VAT was observed in white (p < 0.05) but not African-American (p = 0.57) women. There was no association between the polymorphism and the other phenotypes. These results indicate a significant association between the T265C APOA-II polymorphism and levels of VAT in premenopausal women. This association is present in white but not African-American women. PMID- 15833936 TI - E-selectin genotypes and risk of type 2 diabetes in women. AB - Endothelial dysfunction increases risk for type 2 diabetes. We examined whether variation in the gene for E-selectin (SELE), a biomarker of endothelial dysfunction, was associated with levels of E-selectin or diabetes quantitative traits (including fasting levels of insulin and hemoglobin A(1c)) in 719 nondiabetic participants of the Nurses' Health Study or with risk of diabetes in 602 incident (over 10 years of follow-up) cases and 655 control women matched for age, race, and fasting status. Variation in three single nucleotide polymorphisms previously associated with cardiovascular disease risk and having effects on E selectin function, S128R, G98T, and L554F, was not significantly (p > 0.05) associated with levels of E-selectin or diabetes quantitative traits, or with risk of incident diabetes in the primary analysis. Among women with low levels of subclinical inflammation (C-reactive protein levels below the population median), S128R R allele carriers had a diabetes risk factor-adjusted relative risk of incident diabetes of 1.71 (95% confidence interval, 1.04 to 2.81) relative to those with the SS genotype. Apart from an association in this subgroup, we conclude that the E-selectin variants we examined are not important genetic risk factors for type 2 diabetes in women. PMID- 15833937 TI - Interactive effects between polymorphisms in the beta-adrenergic receptors and longitudinal changes in obesity. AB - We assessed interactions between polymorphisms in the beta-adrenergic receptor genes and longitudinal changes in obesity from childhood to adulthood using longitudinal data collected over a 24-year period from 1973 to 1996. Sex- and age stratified analyses using random coefficients models were used to examine gene gene interaction effects on obesity measures in 1179 African-American and white men and women (71% white, 57% women). Suggestive evidence for an interaction (p = 0.022) between the beta1- and beta2-adrenergic receptors was observed in men for longitudinal change in BMI. Men with Gly/Gly genotypes for both the beta1 and beta2 receptors showed significant increases (approximately 0.6%/yr) in BMI from childhood to adulthood. Women showed suggestive evidence for an interaction (p = 0.035) between the beta1- and beta3-adrenergic receptors for change over time in BMI. Women with Gly/Gly genotypes at the beta1-receptor and carrying at least one beta3-Arg allele showed notable increases in BMI. The regulation of lipolysis and development of obesity differ markedly between men and women and may be influenced by genetic polymorphisms, which contribute to the efficiency of the beta-adrenergic receptors, and hormonal effects on adrenergic receptor activity. PMID- 15833938 TI - Profile of overweight and obese people in a Mediterranean region. AB - OBJECTIVE: To assess the prevalence of excess weight (obesity and overweight) and the related environmental risk factors in a Mediterranean population. Self perception of body weight, attitudes toward weight-control behaviors, and the associated factors were also examined. RESEARCH METHODS AND PROCEDURES: A cross sectional nutritional survey was carried out in the Balearic Islands between 1999 and 2000. A random sample (n = 1200) of the adult population (20 to 60 years old) was interviewed. Dietary questionnaires and a global questionnaire incorporating questions related to sociodemographic and lifestyle variables were utilized. Anthropometric measurements were also obtained. RESULTS: Nearly one-half of the population of the Balearic Islands is above the normal weight range (BMI > 25). The predictors of overweight and obesity in the Balearic Islands are to be over 40 years old, to be married, to form part of the lowest educational levels, and to have a sedentary lifestyle. A large percentage of excess-weight individuals tend to underestimate their BMI, are not concerned about their weight status, tend to snack more often, and have never dieted. DISCUSSION: Although the likely causes of the rise in obesity prevalence are difficult to elucidate from this study, we have identified the profile of individuals with excess weight in the Balearic Islands as well as their attitudes toward their body image. PMID- 15833939 TI - Food viscosity influences caloric intake compensation and body weight in rats. AB - OBJECTIVE: To determine the effects of food viscosity on the ability of rats to compensate for calories in a dietary supplement. RESEARCH METHODS AND PROCEDURES: In a series of four experiments, rats consumed dietary supplements equated for caloric and nutritive content but differing in viscosity. Experiments 1 to 3 examined the ability of the rats to compensate for the calories consumed in low- compared with high-viscosity premeals by reducing intake of a subsequent test meal. Caloric compensation was assessed with a wide range of premeal viscosity levels and with two different non-nutritive thickening agents. Experiment 4 assessed the effects of consuming daily a low-viscosity compared with an equicaloric high-viscosity dietary supplement on longer term body weight gain. RESULTS: Consuming a lower viscosity premeal was followed by significantly more caloric intake (i.e., less caloric compensation) compared with consuming premeals with higher viscosity levels. This effect was not specific to one thickening agent. Furthermore, rats given a low-viscosity supplement daily gained significantly more weight over a 10-week period compared with rats given a high viscosity supplement. DISCUSSION: The results of these experiments suggest that food viscosity may be an important determinant of short-term caloric intake and longer term body weight gain. PMID- 15833940 TI - Adenosinergic modulation of ventilation in obese zucker rats. AB - OBJECTIVES: The goal of our study was to determine whether altered adenosinergic mechanisms contribute to the depressed ventilatory response observed in obese Zucker rats. RESEARCH METHODS AND PROCEDURES: Eight lean and eight obese Zucker rats were studied at 7 to 8 weeks of age. Ventilation (V(E)) during room air, during 5-minute hypercapnic (7% CO(2), balance O(2)), and during 30-minute sustained hypoxic (10% O(2)) exposures were sequentially measured by the barometric method on three separate occasions after the randomized blinded administration of equal volumes of either saline (control), 8-(p-sulfophenyl) theophylline (8-PST, 7 mg/kg, peripheral adenosine antagonist), or aminophylline (AMPH, 15 mg/kg, peripheral and central adenosine antagonist). RESULTS: During room air and hypercapnic exposures, AMPH (but not 8-PST) significantly (p < 0.05) increased V(E) in both lean and obese rats. During acute (2 minute) hypoxic exposure, 8-PST (but not AMPH) significantly depressed V(E) in lean rats. In contrast, AMPH (but not 8-PST) selectively increased V(E) in obese rats. During sustained (10 to 30 minutes) hypoxic exposure, neither AMPH nor 8-PST administration altered V(E) in lean rats. In contrast, AMPH (but not 8-PST) selectively increased V(E) during the late response in obese rats. DISCUSSION: Our findings indicate that obese rats possess altered adenosinergic modulation of ventilatory responses to acute and sustained hypoxia in two opposite ways. We conclude that the reduced hypoxic ventilatory response observed in obese Zucker rats is attributed to depressed adenosinergic peripheral excitatory mechanisms and to enhanced adenosinergic central depression mechanisms, both of which contribute to the blunted ventilatory response in obesity. PMID- 15833941 TI - Mice with low metabolic rates are not susceptible to weight gain when fed a high fat diet. AB - OBJECTIVE: Mice divergently selected for high or low food intake (FI) at constant body mass differ in their resting metabolic rates (RMRs). Low-intake individuals (ML) have significantly lower RMR (by 30%) compared with those from the high intake line (MH). We hypothesized that MLs might, therefore, be more likely to increase their body and fat mass when exposed to a high-fat diet (HFD). RESEARCH METHODS AND PROCEDURES: We exposed both lines to a diet with 44.9% calories from fat for 3 weeks while measuring FI, fecal production, and body mass and then returned the mice to standard chow. RESULTS: When exposed to the HFD, both lines significantly decreased their FI (MH, 40% to 45%; ML, 31% to 35%). This decrease occurred simultaneously with a significant increase in apparent energy absorption efficiency (AEAE). When returned to chow, FI and AEAE returned to the levels observed prior to HFD exposure. Because of the adjustments in FI, the absorbed energy was maintained in the MLs and, thus, body mass remained constant. The MH individuals overcompensated for the elevated energy content and AEAE on the HFD and, therefore, absorbed lower energy than when feeding on chow. These mice also did not significantly change their body mass when on the HFD and must have made adjustments in their energy expenditures. Both lines and both sexes increased in fat content on the HFD, but these effects were not different between lines or sexes. DISCUSSION: We found no support for the hypothesis that mice with low RMRs were more susceptible to weight gain when fed the HFD. PMID- 15833942 TI - Changes in fat mass influence SREBP-1c and UCP-2 gene expression in formerly obese subjects. AB - OBJECTIVE: To investigate the effect of fat mass (FM) reduction on adipose tissue gene expression in terms of lipid synthesis [sterol regulatory binding protein 1c (SREBP-1c)] and lipid oxidation [uncoupling protein 2 (UCP-2)] 2 years after lipid malabsorption and to assess the influence of lipid malabsorption on fat free mass (FFM) maintenance evaluating the expression of genes related to glycolysis [hexokinase (HKII)] and glucose storage [glycogen synthase (GS)]. RESEARCH METHOD AND PROCEDURES: SREBP-1c, UCP-2, HKII, and GS mRNA expression were studied by reverse transcriptase-competitive polymerase chain reaction in 10 massively obese subjects before and 2 years after bilio-pancreatic diversion (BPD). Body composition was assessed by isotopic dilution method and insulin sensitivity by euglycemic-hyperinsulinemic clamp. RESULTS: FM decrease was approximately 60%, whereas FFM remained at normal physiological levels. In adipose tissue, SREBP-1c mRNA reduction (-39%, p < 0.005) was related only to FM changes after BPD, and UCP-2 decrease (-37%, p < 0.05) was dependent on free fatty acid (FFA) changes. No significant variations were observed in HKII and GS gene expression in skeletal muscle. DISCUSSION: Lipid malabsorption induced by BPD altered the expression of genes involved in glucose and lipid metabolism, with different consequences on FM and FFM. The degree of FM loss seems to interfere with SREBP-1c gene suppression to preserve an adequate amount of fat storage, in accordance with the thrifty genotype hypothesis. The reduction of FFAs induced by BPD acts in inhibiting FFA transportation to the mitochondria (UCP-2), contributing to the decreased lipid oxidation inside the adipose tissue. PMID- 15833943 TI - Glucose tolerance and skeletal muscle gene expression in response to alternate day fasting. AB - OBJECTIVE: Alternate day fasting may extend lifespan in rodents and is feasible for short periods in nonobese humans. The aim of this study was to examine the effects of 3 weeks of alternate day fasting on glucose tolerance and skeletal muscle expression of genes involved in fatty acid transport/oxidation, mitochondrial biogenesis, and stress response. RESEARCH METHODS AND PROCEDURES: Glucose and insulin responses to a standard meal were tested in nonobese subjects (eight men and eight women; BMI, 20 to 30 kg/m(2)) at baseline and after 22 days of alternate day fasting (36 hour fast). Muscle biopsies were obtained from a subset of subjects (n = 11) at baseline and on day 21 (12-hour fast). RESULTS: Glucose response to a meal was slightly impaired in women after 3 weeks of treatment (p < 0.01), but insulin response was unchanged. However, men had no change in glucose response and a significant reduction in insulin response (p < 0.03). There were no significant changes in the expression of genes involved in mitochondrial biogenesis or fatty acid transport/oxidation, although a trend toward increased CPT1 expression was observed (p < 0.08). SIRT1 mRNA expression was increased after alternate day fasting (p = 0.01). DISCUSSION: Alternate day fasting may adversely affect glucose tolerance in nonobese women but not in nonobese men. The gene expression results indicate that fatty acid oxidation and mitochondrial biogenesis are unaffected by alternate day fasting. However, the increased expression in SIRT1 suggests that alternate day fasting may improve stress resistance, a commonly observed feature of calorie-restricted rodents. PMID- 15833944 TI - Insulin and endothelin in the acute regulation of adiponectin in vivo in humans. AB - OBJECTIVE: In vitro, insulin and endothelin (ET) both modulate adiponectin secretion from adipocyte cell lines. The current studies were performed to assess whether endogenous ET contributes to the acute action of insulin infusions on adiponectin levels in vivo in humans. RESEARCH METHODS AND PROCEDURES: We studied 17 lean and 20 obese subjects (BMI 21.8 +/- 2.2 and 34.0 +/- 5.0 kg/m(2), respectively). Hyperinsulinemic euglycemic clamp studies were performed using insulin infusion rates of 10, 30, or 300 mU/m(2) per minute alone or with concurrent infusion of BQ123, an antagonist of type A ET receptors. Circulating adiponectin levels were assessed at baseline and after achievement of steady state glucose with the insulin infusion. RESULTS: Adiponectin levels were lower in obese than lean subjects (6.76 +/- 3.66 vs. 8.37 +/- 2.79 microg/mL, p = 0.0148 adjusted for differences across gender). Insulin infusions suppressed adiponectin by a mean of 7.8% (p < 0.0001). In a subset of 13 lean and 14 obese subjects for whom data with and without BQ123 were available, there was no evident effect of BQ123 to modulate clamp-associated suppression of adiponectin (p = 0.16). Surprisingly, there was no evident relationship between steady-state insulin concentrations and adiponectin suppression (r = 0.14, p = 0.30), and again no effect of BQ123 to modify this relationship was seen. DISCUSSION: Despite baseline differences in adiponectin levels, we observed equal suppression of adiponectin with insulin infusions in lean and obese subjects. ET receptor antagonism with BQ123 did not modulate this effect, suggesting that endogenous ET does not have a role in modifying the acute effects of insulin on adiponectin production and/or disposition. PMID- 15833945 TI - Alleles on rat chromosome 4 (D4Got41-Fabp1/Tacr1) regulate subphenotypes of obesity. AB - OBJECTIVE: The use of inbred animal models is an essential component of the genetic dissection of complex diseases. Because quantitative trait loci for serum triglycerides, total cholesterol, and body weight were mapped on chromosome 4 in a cross of BioBreeding/OttawaKarlsburg (BB/OK) and spontaneously hypertensive (SHR) rats, we established a congenic BB.SHR rat strain by introgressing a SHR segment of chromosome 4 (D4Got41-Tacr1) into a BB/OK background. The phenotype of these BB.SHR rats (BB.4S) confirmed the quantitative trait loci. To discover whether the phenotype of BB.4S can only be attributed to the SHR segment per se, we established an additional congenic BB.WOKW strain by introgressing a similar segment of chromosome 4 (D4Got41-Fabp1) of the Wistar Ottawa Karlsburg RT1(u) rat into a BB/OK background, termed briefly BB.4W. RESEARCH METHODS AND PROCEDURES: Male normoglycemic BB/OK (20), BB.4S (20), and BB.4W (16) rats were longitudinally studied for body weight, serum triglycerides, total and high density lipoprotein-cholesterol, and glucose tolerance. At the end of the observation period (32 weeks), serum insulin, leptin, and adiposity index (AI) were determined. RESULTS AND DISCUSSION: Congenic BB.4S and BB.4W were significantly heavier, and AI, serum triglycerides, and total cholesterol values were significantly elevated in BB.4S and BB.4W compared with BB/OK but more pronounced in BB.4S. The highest serum insulin was found in BB.4W and highest leptin in BB.4S. Because the body weight gain and AI were comparable between BB.4S and BB.4W, the obviously higher insulin levels in BB.4W and higher leptin values in BB.4S suggest that the two congenics most probably define two subphenotypes of obesity and provide the unique opportunity to study their genetics. PMID- 15833946 TI - Diet and physical activity behaviors among Americans trying to lose weight: 2000 Behavioral Risk Factor Surveillance System. AB - OBJECTIVE: To examine the prevalence and correlates of trying to lose weight among U.S. adults, describe weight loss strategies, and assess attainment of recommendations for weight control (eating fewer calories and physical activity). RESEARCH METHODS AND PROCEDURES: This study used the Behavioral Risk Factor Surveillance System, a state-based telephone survey of adults > or =18 years of age (N = 184,450) conducted in the 50 states, the District of Columbia, and Puerto Rico in 2000. RESULTS: The prevalence of trying to lose weight was 46% (women) and 33% (men). Women reported trying to lose weight at a lower BMI than did men; 60% of overweight women were trying to lose weight, but men did not reach this level until they were obese. Adults who had a routine physician checkup in the previous year and reported medical advice to lose weight vs. checkup and no medical advice to lose weight had a higher prevalence of trying to lose weight (81% women and 77% men vs. 41% women and 28% men, respectively). The odds of trying to lose weight increased as years of education increased. Among respondents who were trying to lose weight, approximately 19% of women and 22% of men reported using fewer calories and > or =150 min/wk leisure-time physical activity. DISCUSSION: A higher percentage of women than men were trying to lose weight; both sexes used similar weight loss strategies. Education and medical advice to lose weight were strongly associated with trying to lose weight. Most persons trying to lose weight were not using minimum recommended weight loss strategies. PMID- 15833947 TI - Sedentary behavior, physical activity, and the metabolic syndrome among U.S. adults. AB - OBJECTIVE: We examined the associations among physical activity, sedentary behavior, and metabolic syndrome in a representative sample of U.S. adults. RESEARCH METHODS AND PROCEDURES: A total of 1626 men and women > or =20 years old from National Health and Nutrition Examination Survey 1999 to 2000 who attended the morning examination were evaluated. The metabolic syndrome was defined by using the definition from the National Cholesterol Education Program. RESULTS: In unadjusted analysis, participants who did not engage in any moderate or vigorous physical activity during leisure time had almost twice the odds of having metabolic syndrome [odds ratio (OR), 1.90; 95% confidence interval (CI), 1.22 to 2.97] as those who reportedly engaged in > or =150 min/wk of such activity. Adjustment for age, sex, race or ethnicity, educational status, smoking status, and alcohol use attenuated the OR (OR, 1.46; 95% CI, 0.87 to 2.45). Compared with participants who watched television or videos or used a computer <1 h/d outside of work, the adjusted ORs for having metabolic syndrome were 1.41 (95% CI 0.80 to 2.51) for 1 h/d, 1.37 (95% CI 0.85 to 2.20) for 2 h/d, 1.70 (95% CI 0.92 to 3.14) for 3 h/d, and 2.10 (95% CI 1.27 to 3.47) for > or =4 h/d. Additional adjustment for physical activity or sedentary behavior minimally affected the ORs. DISCUSSION: Sedentary behavior is an important potential determinant of the prevalence of the syndrome. Efforts to lessen the amount of time that U.S. adults spend watching television or videos or using a computer, especially if coupled to increases in physical activity, could result in substantial decreases in the prevalence of metabolic syndrome. PMID- 15833948 TI - Effects of exercise on metabolic risk variables in overweight postmenopausal women: a randomized clinical trial. AB - OBJECTIVE: This study examined the effects of exercise on metabolic risk variables insulin, leptin, glucose, and triglycerides in overweight/obese postmenopausal women. RESEARCH METHODS AND PROCEDURES: Sedentary women (n = 173) who were overweight or obese (BMI > or = 25 kg/m(2) or > or =24 kg/m(2) with > or =33% body fat), 50 to 75 years of age, were randomized to 12 months of exercise (> or =45 minutes of moderate-intensity aerobic activity 5 d/wk) or to a stretching control group. Body composition (DXA) and visceral adiposity (computed tomography) were measured at baseline and 12 months. Insulin, glucose, triglycerides, and leptin were measured at baseline and 3 and 12 months. Insulin resistance was evaluated by the homeostasis model assessment formula. Differences from baseline to follow-up were calculated and compared across groups. RESULTS: Exercisers had a 4% decrease and controls had a 12% increase in insulin concentrations from baseline to 12 months (p = 0.0002). Over the same 12-month period, leptin concentrations decreased by 7% among exercisers compared with remaining constant among controls (p = 0.03). Homeostasis model assessment scores decreased by 2% among exercisers and increased 14% among controls from baseline to 12 months (p = 0.0005). The exercise effect on insulin was modified by changes in total fat mass (trend, p = 0.03), such that the exercise intervention abolished increases in insulin concentrations associated with gains in total fat mass. DISCUSSION: Regular moderate-intensity exercise can be used to improve metabolic risk variables such as insulin and leptin in overweight/obese postmenopausal women. These results are promising for health care providers providing advice to postmenopausal women for lifestyle changes to reduce risk of insulin resistance, coronary heart disease, and diabetes. PMID- 15833949 TI - Improved prediction of body fat by measuring skinfold thickness, circumferences, and bone breadths. AB - OBJECTIVE: To develop improved predictive regression equations for body fat content derived from common anthropometric measurements. RESEARCH METHODS AND PROCEDURES: 117 healthy German subjects, 46 men and 71 women, 26 to 67 years of age, from two different studies were assigned to a validation and a cross validation group. Common anthropometric measurements and body composition by DXA were obtained. Equations using anthropometric measurements predicting body fat mass (BFM) with DXA as a reference method were developed using regression models. RESULTS: The final best predictive sex-specific equations combining skinfold thicknesses (SF), circumferences, and bone breadth measurements were as follows: BFM(New) (kg) for men = -40.750 + {(0.397 x waist circumference) + [6.568 x (log triceps SF + log subscapular SF + log abdominal SF)]} and BFM(New) (kg) for women = -75.231 + {(0.512 x hip circumference) + [8.889 x (log chin SF + log triceps SF + log subscapular SF)] + (1.905 x knee breadth)}. The estimates of BFM from both validation and cross-validation had an excellent correlation, showed excellent correspondence to the DXA estimates, and showed a negligible tendency to underestimate percent body fat in subjects with higher BFM compared with equations using a two-compartment (Durnin and Womersley) or a four-compartment (Peterson) model as the reference method. DISCUSSION: Combining skinfold thicknesses with circumference and/or bone breadth measures provide a more precise prediction of percent body fat in comparison with established SF equations. Our equations are recommended for use in clinical or epidemiological settings in populations with similar ethnic background. PMID- 15833950 TI - Altitude exposure should increase serum leptin levels in healthy adults. PMID- 15833964 TI - Preventing dog bites with the help of the Blue Dog. PMID- 15833967 TI - Effects of diets fed to dairy cows before and after calving on their plasma progesterone profiles after calving. AB - Four weeks before their predicted mean second lactation calving date, 60 spring calving Holstein-Friesian cows were blocked into groups of six on the basis of their predicted calving date and body condition score, and allocated at random to one of six dietary treatments in a factorial design: ad libitum grass silage, ad libitum grass silage plus barley straw or ad libitum grass silage plus 3 kg of concentrates, was offered for four weeks before the expected calving date, and after calving they were offered either 4 kg or 8 kg of concentrates plus ad libitum grass silage for eight weeks. On average, the first luteal activity occurred in all the groups at 29 days after calving. Seventeen of the cows had an atypical first plasma progesterone profile; 12 were anovulatory, three had prolonged luteal phases and two became anovulatory after having been cyclic. The cows offered grass silage only before calving had a significantly shorter mean (se) interval between calving and second luteal activity (44.9 [2.1] days), than the cows offered grass silage and straw (53.5 [1.9] days) or grass silage and concentrates (51.5 [3.2] days) (P<0.05). After calving none of the 28 cows offered grass silage and 4 kg of concentrates started cycling before day 21, whereas five of the 30 cows offered grass silage and 8 kg of concentrates cycled before day 21 (P<0.05). PMID- 15833966 TI - Laboratory model to evaluate the role of aerosols in the transport of porcine reproductive and respiratory syndrome virus. AB - The aim of this study was to develop a model to evaluate the aerosol transmission of porcine reproductive and respiratory disease virus (PRRSV). PRRSV (MN 30-100 strain, total dose 3 x 10(6) virus particles) was aerosolised and transported up to 150 m and a portable air sampler was used to collect air samples at 1, 30, 60, 90, 120 and 150 m (five replicates at each distance) and the air samples were tested by TaqMan PCR and virus isolation. The infectivity of the aerosolised PRRSV was tested by exposing six PRRSV-naive pigs for three hours to aerosolised virus that had been transported 150 m. PRRSV RNA was detected in all five replicate air samples collected at 1, 30, 60 and 90 m, in four of the five collected at 120 m, and in three of the five collected at 150 m. Infectious PRRSV was detected by virus isolation at 1 and 30 m (all five replicates), 60, 90 and 120 m (three of the five) and 150 m (two of the five). There was a 50 per cent reduction in the log concentration of PRRSV RNA every 33 m. Three of the six pigs exposed to PRRSV-positive aerosols became infected, and PRRSV RNA was detected in air samples and on swab samples collected from the interior of the chambers that housed the infected pigs while they were being exposed. PMID- 15833968 TI - Diagnosis and treatment of primary intraoral leiomyosarcomas in four dogs. AB - This paper describes the clinical and pathological features, diagnosis and management of primary intraoral leiomyosarcomas in the oral cavity of four dogs; they involved the gingivae of the rostral maxilla and mandible, the mucosa of the upper lip and the soft palate. Their features are compared with the those of leiomyosarcomas commonly encountered in the gastrointestinal tract of dogs. Two of the dogs responded well to surgical treatment; one died after its airways were obstructed by a large palatal tumour; and the fourth was lost to follow-up after its owners declined treatment. PMID- 15833969 TI - Mycoplasma sturni and Mycoplasma gallisepticum in wild birds in Scotland. PMID- 15833970 TI - Alopecia induced by salmonellosis in guinea pigs. PMID- 15833971 TI - Possible exclusion of genotypes in Anaplasma phagocytophilum-infected lambs. PMID- 15833972 TI - First report of avian infectious laryngotracheitis in Morocco. PMID- 15833973 TI - Logos for the RCVS Practice Standards Scheme. PMID- 15833974 TI - Needlestick injuries. PMID- 15833975 TI - BSE agent signatures in a goat. PMID- 15833976 TI - Equine polysaccharide storage myopathy. PMID- 15833977 TI - Teeth, bones and brushing. PMID- 15833978 TI - Assessment of reperfused acute myocardial infarction with two-phase contrast enhanced helical CT: prediction of left ventricular function and wall thickness. AB - PURPOSE: To investigate whether two-phase contrast material-enhanced computed tomographic (CT) findings serve as predictors of changes in left ventricular (LV) function and wall thickness (WT) after acute myocardial infarction (MI) and successful angioplasty. MATERIALS AND METHODS: Ethics committee approval and informed consent were obtained. In 58 patients (51 men and seven women; mean age, 62 years +/- 12 [standard deviation]) who had experienced an acute MI and undergone successful angioplasty, two-phase (acquisitions at 45 seconds and 7 minutes) contrast-enhanced CT was performed in the acute (mean interval between treatment and CT, 37 hours +/- 4) and intermediate (mean interval, 28 days +/- 4) periods and for long-term (mean interval, 12 months +/- 4) follow-up. CT images were reviewed for an early perfusion defect (ED) at 45 seconds and for late enhancement (LE) and a residual perfusion defect (RD) at 7 minutes. Myocardial enhancement patterns and WT were assessed, and LV ejection fraction (LVEF) and percentage decrease in WT were calculated. The patient group was subdivided into three groups according to enhancement pattern: Group 1 included patients with LE but no ED or RD; group 2, patients with ED and LE but no RD; and group 3, patients with ED, LE, and RD. Fisher exact testing was used to measure categorical response. Paired and unpaired t tests were used for comparison between two groups (points); Tukey-Kramer multiple comparison and repeated measures analysis of variance were used for comparisons between the three groups. P < .05 was considered to indicate a significant difference. RESULTS: In group 3 (n = 36), WT in infarcted area was significantly reduced at the intermediate and long-term CT examinations (P < .001). At the intermediate and long-term examinations, percentage decrease in WT was greater in group 2 (n = 10) than in group 1 (n = 12) (P < .05 for intermediate and P < .001 for long-term examination) and was greatest in group 3 (P < .001 for both examinations). LVEF was poorest in group 3 and best in group 1. CONCLUSION: Two-phase contrast enhanced CT proved useful in predicting LV functional recovery and WT in patients who had experienced acute MI and undergone successful angioplasty. PMID- 15833979 TI - Apparent diffusion coefficient of human brain tumors at MR imaging. AB - PURPOSE: To determine if apparent diffusion coefficient (ADC) can be used to differentiate brain tumors at magnetic resonance (MR) imaging. MATERIALS AND METHODS: Institutional review board approval or informed patient consent was not required. MR images were reviewed retrospectively in 275 patients with brain tumors: 147 males and 128 females 1-81 years old, treated between September 1997 and July 2003. Regions of interest were placed manually in tumor regions on MR images, and ADC was calculated with a five-point regression method at b values of 0, 250, 500, 750, and 1000 sec/mm2. ADC values were average values in tumor. All brain tumor subgroups were analyzed. Logistic discriminant analysis was performed by using ADC, age, and patient sex as independent variables to discriminate among tumor groups. RESULTS: A significant negative correlation existed between ADC and astrocytic tumors of World Health Organization grades 2-4 (grade 2 vs grades 3 and 4, accuracy of 91.3% [P < .01]; grade 3 vs 4, accuracy of 82.4% [P < .01]). ADC of dysembryoplastic neuroepithelial tumors (DNTs) was higher than that of astrocytic grade 2 tumors (accuracy, 100%) and other glioneuronal tumors. ADC of malignant lymphomas was lower than that of glioblastomas and metastatic tumors (accuracy, 83.6%; P < .01). ADC of primitive neuroectodermal tumors (PNETs) was lower than that of ependymomas (accuracy, 100%). ADC of meningiomas was lower than that of schwannomas (accuracy, 92.4%; P < .01). ADC of craniopharyngiomas was higher than that of pituitary adenomas (accuracy, 85.2%; P < .05). ADC of epidermoid tumors was lower than that of chordomas (accuracy, 100%). In meningiomas, ADC was not indicative of malignancy grade or histologic subtype. CONCLUSION: ADC is useful for differentiation of some human brain tumors, particularly DNT, malignant lymphomas versus glioblastomas and metastatic tumors, and ependymomas versus PNETs. PMID- 15833980 TI - Assessment of multiple sclerosis lesions with spherical harmonics: comparison of MR imaging and pathologic findings. AB - Spherical harmonics (SH) were used to approximate the volume and three dimensional geometry of multiple sclerosis (MS) lesions in deceased patients. The institutional ethical committee does not require its approval for studies involving pathologic specimens. Pathologic findings were used as the reference standard. In addition, lesion volume was measured with cylindrical approximation (CA). Volumetric comparisons of biases were based on summary statistics, Spearman correlation, Wilcoxon test, and two-way analysis of variance. Shape comparison metrics included mean distance and Dice similarity coefficient (DSC). Eight of 11 lesions had smaller biases with SH method (P < .001). Median biases with SH and CA did not differ significantly, as compared with pathologic findings (r = 1.00 vs 0.99, respectively). Variances of the biases were significantly smaller for SH (P = .04). Ranges of normalized distance and DSC were 0.1%-2.5% and 75%-96%, respectively. Mean DSC was significantly higher than 70% (P < .001). SH method provided unbiased lesion volume and added geometric information that may enable a better understanding of the pathogenesis and lesion evolution over time. PMID- 15833981 TI - Multi-detector row CT systems and image-reconstruction techniques. AB - The introduction in 1998 of multi-detector row computed tomography (CT) by the major CT vendors was a milestone with regard to increased scan speed, improved z axis spatial resolution, and better utilization of the available x-ray power. In this review, the general technical principles of multi-detector row CT are reviewed as they apply to the established four- and eight-section systems, the most recent 16-section scanners, and future generations of multi-detector row CT systems. Clinical examples are used to demonstrate both the potential and the limitations of the different scanner types. When necessary, standard single section CT is referred to as a common basis and starting point for further developments. Another focus is the increasingly important topic of patient radiation exposure, successful dose management, and strategies for dose reduction. Finally, the evolutionary steps from traditional single-section spiral image-reconstruction algorithms to the most recent approaches toward multisection spiral reconstruction are traced. PMID- 15833982 TI - Airway wall thickness in cigarette smokers: quantitative thin-section CT assessment. AB - PURPOSE: To design and validate a dedicated software tool to measure airway dimensions on thin-section computed tomographic (CT) images and to use the tool to prospectively compare airway wall thickness in nonsmokers with normal lung function with that in smokers with and without chronic obstructive pulmonary disease (COPD). MATERIALS AND METHODS: All subjects gave written informed consent. The study was approved by local ethics committee. With Laplacian of Gaussian algorithm, software was tested in phantom and excised sheep lung fixed in inflation and validated with Bland-Altman analysis. Study prospectively included nine nonsmokers (six women, three men; mean age, 53 years +/- 5.6 [standard error of the mean]) with normal lung function (group 1), seven smokers (three women, four men; mean age, 56 years +/- 5.6) with normal lung function (group 2), and eight smokers (zero women, eight men; mean age, 65 years +/- 4.0) with COPD. Calculations were determined with spirometrically gated CT: For each selected bronchus, the wall area (WA), internal area (IA), airway caliber (sum of IA and WA), and WA/IA ratio were calculated. For each patient, summation of WA to summation of IA (SigmaWA/SigmaIA) ratio, which reflected normalized airway wall thickness, was calculated. Groups were compared by using analysis of variance with generalized linear model and unpaired t test. Pearson correlation coefficient was used to assess correlation between software measurements and pulmonary function test results. RESULTS: Comparison of measurements in phantom and excised sheep lung with algorithm measurements revealed that the latter were reliable and repeatable. In clinical study, SigmaWA/SigmaIA ratio was significantly different among three groups (P < .001). Normalized airway wall thickness and IA were significantly related to lung function test data, including forced expiratory volume in 1 second (r = -0.54, P = .006), specific airway conductance (r = -0.45, P = .03), and forced expiratory flow between 25% and 75% of vital capacity (r = -0.65, P < .001). CONCLUSION: This software provides accurate and reproducible measurements of IA and WA of bronchi on thin-section CT images and demonstrates that in vivo normalized airway wall thickness was larger in smokers with COPD than it was in smokers or nonsmokers without COPD. PMID- 15833983 TI - The pituitary gland: changes on MR images during the 1st year after delivery. AB - PURPOSE: To longitudinally and prospectively investigate changes in the volume and signal intensity on T1-weighted magnetic resonance (MR) images of the pituitary gland up to 1 year after delivery and evaluate whether termination of lactation has an effect on these parameters. MATERIALS AND METHODS: All participants provided informed consent for participation in the study, which was approved by the institutional review board. Thirteen volunteers (mean age, 28 years; age range, 26-32 years) underwent MR imaging 2 and 4 weeks after delivery and then at intervals of 0.5-2.0 months until 1 year after delivery. Eight participants terminated lactation during the study period. Sagittal and coronal T1-weighted images were obtained. Signal intensities of the anterior and posterior lobes of the pituitary were calculated relative to that of the pons. The volume of the pituitary was also calculated. Two-tailed paired Student t tests and separate simple linear regression analyses were used to test for statistically significant differences. RESULTS: The mean pituitary volume was 544 mm3 at 2 weeks, 523 mm3 at 4 months, 512 mm3 at 8 months, and 511 mm3 at 12 months after delivery, with significant differences between 2 weeks and 4 months (P = .002) and between 4 and 8 months (P = .003) after delivery. The mean ratio of the signal intensity of the anterior lobe of the pituitary to the signal intensity of the pons was 1.11 at 2 weeks, 1.07 at 4 months, 1.03 at 8 months, and 1.00 at 12 months after delivery, with significant differences between 2 weeks and 4 months (P = .004) and between 4 and 8 months (P = .0001) after delivery. Termination of lactation had no statistically significant effect on pituitary volume or the ratio of the signal intensity of the anterior or posterior lobe of the pituitary to the signal intensity of the pons. CONCLUSION: The volume of the pituitary gland decreases up to 8 months after delivery, and the T1-weighted signal intensity of the anterior lobe of the pituitary decreases; termination of lactation has no statistically significant effect on these parameters. PMID- 15833984 TI - Anomalous coronary arteries in adults: depiction at multi-detector row CT angiography. AB - PURPOSE: To retrospectively determine the imaging features of anomalous coronary arteries depicted at multi-detector row computed tomographic (CT) angiography in 18 patients seen at four institutions. MATERIALS AND METHODS: Eighteen patients underwent imaging with a four- or 16-section multi-detector row CT unit by using retrospective electrocardiographic (ECG) gating after infusion of 120-150 mL of intravenous contrast material. Section thicknesses of 0.8-3.0 mm were achieved during breath holding, and images were reconstructed with a 50% overlap. Volumetric reconstructions were obtained for each patient. Each study was assessed retrospectively for the origin and course of the anomalous coronary artery by two thoracic radiologists; decisions were made in consensus. Institutional review board exemption and informed consent waiver was granted at each institution. The study was compliant with the Health Insurance Portability and Accountability Act. RESULTS: Seventeen patients were referred because of equivocal findings at cardiac catheterization or echocardiography; in one, the anomalous coronary artery was incidental. A total of 20 anomalous vessels were found. Twelve patients with 14 variant vessels had an anomalous origin of a left coronary artery (right cusp, 13; noncoronary cusp, one). In four patients, an anomalous right coronary artery originated from the left side; one patient had a single coronary artery arising from the right cusp. In one patient, a left coronary artery-to-vein fistula was observed. In 10 patients, the anomalous vessel passed between the aorta and the main pulmonary artery or right ventricular outflow track. In each case, the origin of the anomalous coronary artery and its course in relationship to the great vessels were unequivocally demonstrated. Volumetric images were useful for showing the three-dimensional orientation of the anomalous coronary artery with respect to the great vessels and cardiac chambers. CONCLUSION: Multi-detector row CT angiography provided accurate depiction of vessel origin and course in this review of 20 anomalous coronary arteries. The results of this study suggest that CT is a viable noninvasive modality for delineating coronary arterial anomalies, particularly if findings at coronary angiography are equivocal. PMID- 15833985 TI - Gastrointestinal stromal tumor: new nodule-within-a-mass pattern of recurrence after partial response to imatinib mesylate. AB - PURPOSE: To investigate a new pattern of tumor recurrence observed at imaging in patients with metastatic gastrointestinal stromal tumor (GIST) after initial partial response to imatinib mesylate. MATERIALS AND METHODS: Ninety-two patients with metastatic GIST who underwent treatment in a clinical trial with oral imatinib mesylate were followed up for 29 months. An institutional review board approved protocol was used. The study complied with the Health Insurance Portability and Accountability Act, and written informed consent was obtained from all patients. Images of the chest, abdomen, and pelvis, acquired with computed tomography (CT), positron emission tomography (PET), and, in some cases, magnetic resonance imaging, were evaluated for treatment response and disease recurrence. Thirty-nine patients (29 men, 10 women; age range, 18-84 years; mean, 49.2 years) had recurrent disease after an initial variable period of response (range, 2-24 months; median, 14.4 months). Initial response was determined with findings of decreased uptake of fluorine 18 fluorodeoxyglucose at PET, shrinkage of tumor, and decreased attenuation at CT. Images were evaluated for disease recurrence by two experienced radiologists who were blinded to each other's interpretation but not to clinical details. Final reading was performed by consensus. RESULTS: A nodule within a mass was seen in 21 of 39 patients (in intrahepatic tumor [n = 8], extrahepatic tumor [n = 10], or both intra- and extrahepatic tumors [n = 3]) and was the first sign of disease progression in 17 of 21 patients. Other patterns of recurrence included new site of disease (n = 7), regrowth of preexistent lesion (n = 20), and mixed (more than one) pattern (n = 9). Disease progression was verified at needle biopsy (n = 16), follow-up imaging (n = 14), and/or surgical resection (n = 9). CONCLUSION: A nodule within a mass is an important sign of recurrent GIST, but measurements of overall tumor size may not enable detection of such nodules. PMID- 15833986 TI - Attenuation of acute and chronic pulmonary emboli. AB - PURPOSE: To compare retrospectively the attenuation coefficients of acute and chronic pulmonary embolism (PE). MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was waived. The study was compliant with requirements of the Health Insurance Portability and Accountability Act. All patients with chronic PE, from July 2001 to January 2004, were identified via a radiology report search system; of the 39 identified, 25 were excluded because the thrombi were too small to measure or were obscured by streak artifacts or because there was no corroborative evidence of chronic PE. Of 27 consecutive patients with acute PE who were also identified, two were excluded because of streak artifacts. The final study group included six women and eight men with chronic PE (mean age, 50 years; range, 26-76 years) and 11 women and 14 men with acute PE (mean age, 61 years; range, 33-83 years) (P = .01 for age). Images were acquired with a four-detector row computed tomographic scanner and 1.25-mm collimation. Two readers made independent attenuation measurements of the largest thrombus in each patient at a workstation. Statistical analysis included calculation of means and standard deviations, the t test, and the Bland-Altman test. RESULTS: Reader 1 found mean attenuation of 90 HU +/- 30 (range, 54-155 HU) for chronic PE and 33 HU +/- 15 (range, 6-63 HU) for acute PE (P < .001). Reader 2 found mean attenuation measurements of 83 HU +/- 32 (range, 32-135 HU) for chronic PE and 33 HU +/- 14 (range, 13-65 HU) for acute PE (P < .001). The mean attenuation for both readers was 33 HU for acute PE (95% confidence interval: 26, 41 HU) and 87 HU for chronic PE (95% confidence interval: 66, 107 HU). The Bland Altman test demonstrated agreement between readers. CONCLUSION: The mean attenuation measurement in chronic PE is significantly higher than in acute PE. PMID- 15833987 TI - Living donor candidates for right hepatic lobe transplantation: evaluation at CT cholangiography--initial experience. AB - PURPOSE: To retrospectively evaluate computed tomographic (CT) cholangiography in the depiction of second-order biliary tract anatomy in living donor candidates for right hepatic lobe transplantation. MATERIALS AND METHODS: Human research committee approval was obtained, informed consent was not required, and the study was compliant with the Health Insurance Portability and Accountability Act. The authors identified all living right-lobe liver donor candidates who underwent CT cholangiography at their institution between October 2001 (when CT cholangiography was introduced at the institution) and March 2003 (n = 62). There were 41 men (mean age, 36 years; range, 18-55 years) and 21 women (mean age, 40 years; range, 22-55 years). Two readers in consensus rated quality of second order bile duct visualization at CT cholangiography on a four-point scale (0, not seen; 3, excellent visualization) and noted the presence of variant second-order biliary tract branching anatomy. CT cholangiography findings were compared with those at surgery in subjects who underwent right hepatic lobe retrieval (n = 24). In addition, adult donors who underwent right hepatic lobe retrieval between January 2000 and March 2003 (29 men, mean age, 35 years [range, 20-52 years]; 18 women, mean age, 38 years [range, 23-54 years]) were identified. Numbers of donors who underwent intraoperative cholangiography before and after the introduction of CT cholangiography were compared by using the Fisher exact test. RESULTS: The mean second-order bile duct score at CT cholangiography was 2.9 (range, 2-3). Of 24 subjects who underwent right lobe retrieval, biliary tract anatomy determined at CT cholangiography was concordant with findings at surgery in 23 (96%). Variant second-order branching anatomy was seen in 13 subjects (54%) at surgery; one variant branch was missed at CT cholangiography. Of 47 subjects who underwent right hepatic lobe retrieval, significantly fewer subjects required conventional intraoperative cholangiography after the introduction of CT cholangiography (three of 24 subjects [12%]) than before (23 of 23 subjects; P < .0001). CONCLUSION: CT cholangiography accurately depicts biliary tract anatomy in living donor candidates for right hepatic lobe transplantation, and donors who undergo preoperative CT cholangiography are unlikely to need conventional intraoperative cholangiography. PMID- 15833988 TI - Diagnostic imaging costs: are they driving up the costs of hospital care? AB - PURPOSE: To retrospectively determine how changes in utilization of computed tomography (CT), magnetic resonance (MR) imaging, and other imaging technologies between 1996 and 2002 influenced costs of inpatient hospital care at one large academic medical center. MATERIALS AND METHODS: Institutional review board did not require its approval or patient informed consent for studies with use of billing data. Patient anonymity was protected by removal of potentially identifying information. Data on hospital costs for 17 139 patients admitted to Massachusetts General Hospital, Boston, Mass, between 1996 and 2002 were downloaded from hospital cost-accounting system; sample was restricted to inpatients with diagnoses in diagnosis-related groups 014-015 (Stroke and TIA [transient ischemic attack]), 164-167 (Appendectomy), 082 (Lung Cancer), 182-183 (Upper Gastrointestinal Conditions), 148-149 (Colon Cancer), and 243 (Back Problems). For each patient, data on demographics, all products and services used, and costs associated with each product or service were obtained. By using institutional codes, we calculated costs of CT, MR imaging, and total imaging relative to total hospital costs. Statistical analyses were performed with Student t test and multiple linear regression analysis. RESULTS: Between 1996 and 2002, number of inpatient CT and MR images obtained at the hospital more than doubled. In 2002, hospital costs were 155% those of 1996 levels; inpatient imaging costs were 151% those of 1996 levels. Total costs increased an average of 7.8% per year; imaging costs increased 8.3% per year. Although highly variable over the study period, as a percentage of total imaging costs, CT and MR imaging costs appeared to remain stable relative to costs of other imaging modalities. CONCLUSION: Despite substantial increases in utilization of inpatient CT, MR imaging, and other imaging technologies, diagnostic imaging costs increased at approximately same rate as did total costs for inpatients with several diagnoses. CT and MR imaging do not appear to be driving the cost increases seen between 1996 and 2002. PMID- 15833989 TI - Multi-detector row CT in evaluation of 94 living renal donors by readers with varied experience. AB - PURPOSE: To retrospectively assess the accuracy of four-section multi-detector row computed tomography (CT) in the evaluation of renal transplant donors when scans are read by one of multiple readers with varied levels of expertise, by using surgery as the reference standard. MATERIALS AND METHODS: This retrospective study was approved by the institutional review board and complied with the Health Insurance Portability and Accountability Act. Informed consent was waived. Between October 1999 and March 2003, 94 renal donors (42 men, 52 women; mean age, 44 years) underwent four-section multi-detector row CT. Unenhanced scanning of the abdomen was performed with 5-mm section thickness and table speed of 15 mm per rotation. Next, 135-150 mL of nonionic iodinated (300 mg/mL) contrast material was injected intravenously at a rate of 4-5 mL/sec. Contrast material-enhanced CT was initiated 20-25 seconds, 65-70 seconds, and 10 minutes after start of injection. Arterial phase scanning was performed with 1.25 mm section thickness and 7.5-mm table speed. Venous and excretory phase scanning was performed with 2.5-mm section thickness and 15-mm table speed. Each scan was evaluated independently by one of 11 readers for renal vascular and ureteral anatomic variants. Findings at CT were compared with those at surgery. Sensitivity and specificity (with 95% confidence intervals) and accuracy of CT were calculated on the basis of presence or absence of variant anatomy at surgery. RESULTS: CT depicted 107 of 114 renal arteries confirmed at surgery; seven accessory arteries were missed in six donor kidneys. CT depicted 95 of 98 renal veins confirmed at surgery. Sensitivity and specificity of CT were 66% and 100%, 75% and 100%, and 50% and 100%, and overall accuracy was 94%, 97%, and 99%, for identification of variant anatomy of renal arteries, veins, and ureters, respectively. CONCLUSION: Multi-detector row CT as the sole imaging technique in the preoperative evaluation of living renal donors is accurate even when images are read by multiple readers with varied levels of expertise. PMID- 15833990 TI - Achalasia with complete relaxation of lower esophageal sphincter: radiographic manometric correlation. AB - PURPOSE: To evaluate retrospectively the presence of complete relaxation of the lower esophageal sphincter (LES) at manometry in patients with achalasia depicted on barium esophagograms. MATERIALS AND METHODS: The institutional review board approved this retrospective study and did not require informed consent. A search of radiology and manometry records identified 21 patients (12 men, nine women; mean age, 52.4 years) with achalasia depicted on barium esophagograms who had undergone manometric examinations and met the inclusion criteria. Radiologic reports and images were reviewed for presence or absence of primary peristalsis, impaired LES opening, esophageal dilatation, delayed emptying of barium, and nonperistaltic contractions. Manometry reports were reviewed for presence or absence of peristalsis or simultaneous esophageal contractions. Resting and residual LES pressures were recorded to determine whether LES relaxation was complete or incomplete. Medical records were reviewed to determine clinical presentation and follow-up (treatment and patient course), and radiographic files were reviewed to determine radiographic findings at follow-up examinations. Clinical characteristics (eg, age, dysphagia, and weight loss) were correlated with LES relaxation at manometry. Data were analyzed statistically with Fisher exact and Wilcoxon rank sum testing. RESULTS: All 21 patients with radiographic findings of achalasia had aperistalsis at manometry. Fourteen patients (67%) had incomplete LES relaxation at manometry during swallowing, and seven (33%) had complete LES relaxation. There were no significant differences between patients with complete LES relaxation and those with incomplete LES relaxation in mean age (P = .59), duration of dysphagia (P = .18), or weight loss (P > .99). Clinical follow-up findings were available for six patients with complete LES relaxation at manometry and 10 with incomplete relaxation. Symptoms resolved after treatment in all six patients with complete LES relaxation. Six (60%) of 10 patients with incomplete LES relaxation had resolution of symptoms after treatment, and four (40%) had substantial improvement. CONCLUSION: These data suggest that in patients with typical radiographic findings of achalasia, the barium study can be used to guide treatment without a need for manometry. If radiographic findings are equivocal, however, manometry may be required for a more certain diagnosis. PMID- 15833991 TI - Multistage ethanol sclerotherapy of soft-tissue arteriovenous malformations: effect on pulmonary arterial pressure. AB - PURPOSE: To retrospectively investigate how repeat injections of absolute ethanol in therapeutic doses, required for multisession sclerotherapy of large high-flow soft-tissue arteriovenous malformations (AVMs) in patients with normal cardiopulmonary function, affect pulmonary arterial pressure (PAP). MATERIALS AND METHODS: Study received approval and waiver of informed consent by institutional review board and was conducted in 16 male and 16 female patients with AVMs who underwent repeat sclerotherapy (142 sessions total) with absolute ethanol from July 1997 to December 2003. PAPs were monitored during first session in all patients. In subsequent sessions, PAP was monitored with pulmonary catheter when predicted single dose of ethanol exceeded 3 mL and total amount exceeded 0.25 mL/kg. PAP was measured in 104 sessions. Serum ethanol levels from blood samples obtained at end of each session were reviewed. PAP parameters were analyzed at beginning and end of each session, and highest value was recorded to assess any increase after repeat therapy. Difference between initial and highest PAP values recorded in a session (Delta(max)) was noted to determine any increase during repeat sessions. Possible relationship was reviewed between this value and amount of ethanol used. For sessions without PAP monitoring, mixed model was used for statistical analysis. RESULTS: Total ethanol used was variable. In 43 sessions, highest mean PAP exceeded 25 mm Hg. Incidence of sustained pulmonary hypertension (mean PAP > 25 mm Hg) at end of each session was 30.8% (32 of 104 sessions). Initial PAP parameters did not increase or decrease during repeat sessions. No significant changes in Delta(max) of systolic and mean PAP were observed with increasing number of sessions. Rather, Delta(max) of diastolic PAP was reduced after repeat sessions (P = .03). There was no significant correlation between serum ethanol level and PAP parameters at end of sessions. Relationships between Delta(max) values of systolic, mean, and diastolic PAP and total ethanol used were not significant. CONCLUSION: High incidence of acute pulmonary hypertension was observed in each sclerotherapy session without lasting effect on PAP. PMID- 15833992 TI - Hepatic lesions deemed too small to characterize at CT: prevalence and importance in women with breast cancer. AB - PURPOSE: To retrospectively evaluate the prevalence and clinical importance of hepatic lesions considered too small to characterize (TSTC) at initial computed tomography (CT) in women with breast cancer. MATERIALS AND METHODS: Approval for this retrospective study was obtained from the institutional review board, which waived the requirement for informed consent. For each woman who received a diagnosis of breast cancer between 1998 and 2002, the authors reviewed the report of the first contrast material-enhanced CT examination that included assessment of the liver. For women with no definite liver metastasis and at least one hepatic lesion considered TSTC, reports of follow-up imaging examinations were reviewed for a change in lesion size; medical records and images were reviewed if there was a change in lesion size. The 95% confidence intervals (CIs) were calculated for best- and worst-case analyses of cases in which different assumptions were used to classify a lesion as benign. RESULTS: Of 7692 women, 1012 (13.2%) underwent contrast-enhanced CT including liver assessment. The mean age of the 1012 women was 54.6 years (range, 20.7-89.1 years). The median time from diagnosis of breast cancer to initial CT examination was 14.1 weeks (range, 3.7 to 296 weeks). The presence of at least one hepatic lesion deemed TSTC was reported in 277 of 941 women (29.4%) in whom no definite hepatic metastasis was reported. Subsequent imaging examinations were performed in 191 of the 277 women (69.0%) (median time from initial CT to last follow-up imaging examination, 54 weeks; range, 0.3-302 weeks). Those examinations revealed the lesions were unchanged in 175 (91.6%) women, no longer visible in eight (4.2%), and larger in six (3.1%). In two women (1.0%), change could not be determined. The enlarging hepatic lesions deemed TSTC represented metastatic breast cancer (three patients), metastatic pancreatic cancer (one patient), or cysts (one patient); in one patient, the etiology was not known. Results of best- and worst-case analyses showed that the lesions were benign in 96.9% (95% CI: 93%, 99%) and 92.7% (95% CI: 88%, 96%) of women, respectively. CONCLUSION: In 92.7%-96.9% of women with breast cancer and hepatic lesions deemed TSTC but no definite liver metastases at initial CT, the lesions represented a benign finding. PMID- 15833993 TI - Acute appendicitis: added diagnostic value of coronal reformations from isotropic voxels at multi-detector row CT. AB - PURPOSE: To assess retrospectively the added value of coronal reformations from isotropic voxels obtained with 16-section multi-detector row computed tomography (CT) of the abdomen and pelvis in patients with suspected acute appendicitis. MATERIALS AND METHODS: This study was approved by the institutional review board, and informed consent was waived. One hundred consecutive patients (21 men, 79 women; mean age, 38 years) with suspected appendicitis underwent 16-section multi detector row CT (section thickness, 0.625 mm; pitch, 1.75; table speed, 35 mm/sec [17.5 mm per rotation, two rotations]; and gantry speed, 0.5 second per rotation), with coronal reformations. Twenty-four patients had appendicitis; 76 did not. Protocol included 150 mL oral iopamidol administered at 3 mL/sec. Transverse scans were reconstructed with 5-mm-thick sections at 5-mm intervals and 0.625-mm-thick sections at 0.625-mm intervals. The second data set was reformatted coronally, with 3-mm-thick sections at 5-mm intervals. Three independent blinded readers interpreted transverse scans alone and then coronal scans; confidence in visualization of any portion of appendix, entire appendix, wall thickening, distention, inflammation, fluid, and appendicitis was scored with 1-5 scale. Sensitivity and specificity were determined for each reader and compared by means of signed rank test. Agreement between readers was determined with kappa statistic. Differences in mean confidence ratings for each finding were determined with Wilcoxon signed rank test. RESULTS: Mean sensitivity and specificity for all three readers together were 96% and 95% for transverse reformations alone and 95% and 94% for combined transverse and coronal reformations (not significant), respectively. Visualization rates for portion or all of appendix were higher for combined transverse and coronal reformations than for transverse reformations alone (higher mean confidence scores: 0.23 higher [P < .009] and 0.51 higher [P < .001], respectively). In patients without appendicitis, transverse and coronal reformations together enhanced confidence in exclusion of wall thickening, distention, and fluid (lower confidence scores: 0.21 lower [P < .001], 0.17 lower [P < .01], 1.00 lower [P < .001], respectively). Combined transverse and coronal reformations enhanced confidence in identification of appendix in mean of 57 patients. Combined transverse and coronal scans helped exclude appendicitis in mean of 38 patients and aided diagnosis of it in 15. CONCLUSION: Sixteen-section multi-detector row CT transverse and coronal reformations are equally sensitive and specific for diagnosis of appendicitis. Coronal reformations improve confidence in visualization of appendix (whether diseased or normal) and in diagnosis or exclusion of appendicitis. PMID- 15833994 TI - Micrometer-sized supported lipid bilayer arrays for bacterial toxin binding studies through total internal reflection fluorescence microscopy. AB - In this article, we present the use of micron-sized lipid domains, patterned onto planar substrates and within microfluidic channels, to assay the binding of bacterial toxins via total internal reflection fluorescence microscopy. The lipid domains were patterned using a polymer lift-off technique and consisted of ganglioside-populated distearoylphosphatidylcholine:cholesterol supported lipid bilayers (SLBs). Lipid patterns were formed on the substrates by vesicle fusion followed by polymer lift-off, which revealed micron-sized SLBs containing either ganglioside G(T1b) or G(M1). The ganglioside-populated SLB arrays were then exposed to either cholera toxin B subunit or tetanus toxin C fragment. Binding was assayed on planar substrates by total internal reflection fluorescence microscopy down to 100 pM concentration for cholera toxin subunit B and 10 nM for tetanus toxin fragment C. Apparent binding constants extracted from three different models applied to the binding curves suggest that binding of a protein to a lipid-based receptor is influenced by the microenvironment of the SLB and the substrate on which the bilayer is formed. Patterning of SLBs inside microfluidic channels also allowed the preparation of lipid domains with different compositions on a single device. Arrays within microfluidic channels were used to achieve segregation and selective binding from a binary mixture of the toxin fragments in one device. The binding and segregation within the microfluidic channels was assayed with epifluorescence as proof of concept. We propose that the method used for patterning the lipid microarrays on planar substrates and within microfluidic channels can be easily adapted to proteins or nucleic acids and can be used for biosensor applications and cell stimulation assays under different flow conditions. PMID- 15833995 TI - Keeping lung surfactant where it belongs: protein regulation of two-dimensional viscosity. AB - Lung surfactant causes the surface tension, gamma, in the alveoli to drop to nearly zero on exhalation; in the upper airways gamma is approximately 30 mN/m and constant. Hence, a surface tension gradient exists between alveoli and airways that should lead to surfactant flow out of the alveoli and elimination of the surface tension gradient. However, the lung surfactant specific protein SP-C enhances the resistance to surfactant flow by regulating the ratio of solid to fluid phase in the monolayer, leading to a jamming transition at which the monolayer transforms from fluidlike to solidlike. The accompanying three orders of magnitude increase in surface viscosity helps minimize surfactant flow to the airways and likely stabilizes the alveoli against collapse. PMID- 15833996 TI - Dual binding modes for an HMG domain from human HMGB2 on DNA. AB - High mobility group B (HMGB) proteins contain two HMG box domains known to bind without sequence specificity into the DNA minor groove, slightly intercalating between basepairs and producing a strong bend in the DNA backbone. We use optical tweezers to measure the forces required to stretch single DNA molecules. Parameters describing DNA flexibility, including contour length and persistence length, are revealed. In the presence of nanomolar concentrations of isolated HMG box A from HMGB2, DNA shows a decrease in its persistence length, where the protein induces an average DNA bend angle of 114 +/- 21 degrees for 50 mM Na+, and 87 +/- 9 degrees for 100 mM Na+. The DNA contour length increases from 0.341 +/- 0.003 to 0.397 +/- 0.012 nm per basepair, independent of salt concentration. In 50 mM Na+, the protein does not unbind even at high DNA extension, whereas in 100 mM Na+, the protein appears to unbind only below concentrations of 2 nM. These observations support a flexible hinge model for noncooperative HMG binding at low protein concentrations. However, at higher protein concentrations, a cooperative filament mode is observed instead of the hinge binding. This mode may be uniquely characterized by this high-force optical tweezers experiment. PMID- 15833997 TI - Water-soluble hybrid nanoclusters with extra bright and photostable emissions: a new tool for biological imaging. AB - We report the generation of a previously unknown class of water-soluble organic inorganic hybrid nanoclusters composed of silver and thioflavin T with remarkable fluorescent properties. These hybrid nanoclusters display an extra bright fluorescence in aqueous solutions without any detectable photobleaching. Furthermore, the fluorescent nanoclusters can be generated in situ by sensitized photoreduction of Ag+. Organic-inorganic nanoclusters have remarkable advantages over the known classes of fluorescent probes for the development of ultrasensitive biological assays, cell imaging, or studies of single molecules. As an example of a practical biological application, imaging of amyloid fibrils produced from recombinant mammalian prion proteins and nonprion proteins using hybrid nanoclusters is presented. PMID- 15833998 TI - Rigid-cluster models of conformational transitions in macromolecular machines and assemblies. AB - We present a rigid-body-based technique (called rigid-cluster elastic network interpolation) to generate feasible transition pathways between two distinct conformations of a macromolecular assembly. Many biological molecules and assemblies consist of domains which act more or less as rigid bodies during large conformational changes. These collective motions are thought to be strongly related with the functions of a system. This fact encourages us to simply model a macromolecule or assembly as a set of rigid bodies which are interconnected with distance constraints. In previous articles, we developed coarse-grained elastic network interpolation (ENI) in which, for example, only Calpha atoms are selected as representatives in each residue of a protein. We interpolate distance differences of two conformations in ENI by using a simple quadratic cost function, and the feasible conformations are generated without steric conflicts. Rigid-cluster interpolation is an extension of the ENI method with rigid-clusters replacing point masses. Now the intermediate conformations in an anharmonic pathway can be determined by the translational and rotational displacements of large clusters in such a way that distance constraints are observed. We present the derivation of the rigid-cluster model and apply it to a variety of macromolecular assemblies. Rigid-cluster ENI is then modified for a hybrid model represented by a mixture of rigid clusters and point masses. Simulation results show that both rigid-cluster and hybrid ENI methods generate sterically feasible pathways of large systems in a very short time. For example, the HK97 virus capsid is an icosahedral symmetric assembly composed of 60 identical asymmetric units. Its original Hessian matrix size for a Calpha coarse-grained model is >(300,000)(2). However, it reduces to (84)(2) when we apply the rigid-cluster model with icosahedral symmetry constraints. The computational cost of the interpolation no longer scales heavily with the size of structures; instead, it depends strongly on the minimal number of rigid clusters into which the system can be decomposed. PMID- 15833999 TI - Equilibrium and kinetic folding pathways of a TIM barrel with a funneled energy landscape. AB - The role of native contact topology in the folding of a TIM barrel model based on the alpha-subunit of tryptophan synthase (alphaTS) from Salmonella typhimurium (Protein Data Bank structure 1BKS) was studied using both equilibrium and kinetic simulations. Equilibrium simulations of alphaTS reveal the population of two intermediate ensembles, I1 and I2, during unfolding/refolding at the folding temperature, Tf = 335 K. Equilibrium intermediate I1 demonstrates discrete structure in regions alpha0-beta6 whereas intermediate I2 is a loose ensemble of states with N-terminal structure varying from at least beta1-beta3 (denoted I2A) to alpha0-beta4 at most (denoted I2B). The structures of I1 and I2 match well with the two intermediate states detected in equilibrium folding experiments of Escherichia coli alphaTS. Kinetic folding simulations of alphaTS reveal the sequential population of four intermediate ensembles, I120Q, I200Q, I300Q, and I360Q, during refolding. Kinetic intermediates I120Q, I200Q, and I300Q are highly similar to equilibrium alphaTS intermediates I2A, I2B, and I1, respectively, consistent with kinetic experiments on alphaTS from E. coli. A small population (approximately 10%) of kinetic trajectories are trapped in the I120Q intermediate ensemble and require a slow and complete unfolding step to properly refold. Both the on-pathway and off-pathway I120Q intermediates show structure in beta1-beta3, which is also strikingly consistent with kinetic folding experiments of alphaTS. In the off-pathway intermediate I(120Q), helix alpha2 is wrapped in a nonnative chiral arrangement around strand beta3, sterically preventing the subsequent folding step between beta3 and beta4. These results demonstrate the success of combining kinetic and equilibrium simulations of minimalist protein models to explore TIM barrel folding and the folding of other large proteins. PMID- 15834000 TI - Resolution of two substrate-binding sites in an engineered cytochrome P450eryF bearing a fluorescent probe. AB - To elucidate the mechanisms of cooperativity of cytochrome P450eryF an SH reactive fluorescent probe was introduced close to the substrate-binding site. Cys-154, the only accessible cysteine, was eliminated by site-directed mutagenesis, and a novel cysteine was substituted for Ser-93 in the B'/C loop. S93C, C154A, C154S, S93C/C154A, and S93C/S154C were characterized in terms of affinity for 1-pyrenebutanol (1-PB), cooperativity, and ionic-strength dependence of the 1-PB-induced spin shift. S93C/C154S retains the key functional properties of the wild-type, and modification by three different SH-reactive probes had little effect on the characteristics of the enzyme. The labeled proteins exhibited fluorescence resonance energy transfer from 1-PB to the label, which allowed us to resolve two substrate-binding events, and to determine the corresponding KD values (KD1 = 1.2 +/- 0.2 microM, KD2 = 9.4 +/- 0.8 microM). Using these values for analysis of the substrate-induced spin transition, we demonstrate that the interactions of P450eryF with 1-PB are consistent with a sequential binding mechanism, where substrate interactions at a higher-affinity site cause a conformational transition crucial for the binding of the second substrate molecule and subsequent spin shift. This transition is apparently associated with an important rearrangement of the system of salt links in the proximity of Cys-154. PMID- 15834001 TI - Local compressibilities of proteins: comparison of optical experiments and simulations for horse heart cytochrome-c. AB - Spectroscopy with probe molecules yields local information on the environment of the probe. In this article we compare local compressibilities of cytochrome-c as obtained from molecular dynamics simulations with experimental results as obtained from spectroscopic measurements. The simulations show that the protein core around the heme is much less compressible in a glycerol/water solvent than in pure water. The pocket is also much less compressible than the protein as a whole, although the compressibility of the water inside the rather incompressible protein-core is almost liquidlike. We show that the local compressibility values capture the collective correlations of local volume fluctuations with volume fluctuations in the surrounding protein-solvent system. The decoupling of the volume fluctuations of the core from the solvent shell explains the reduction of the heme-core-compressibility in glycerol/water solvent. This decoupling could be traced back to the suppression of the exchange between pocket-water and hydration shell-water upon addition of glycerol as co-solvent. PMID- 15834002 TI - Intra- and inter-islet synchronization of metabolically driven insulin secretion. AB - Insulin secretion from pancreatic beta-cells is pulsatile with a period of 5-10 min and is believed to be responsible for plasma insulin oscillations with similar frequency. To observe an overall oscillatory insulin profile it is necessary that the insulin secretion from individual beta-cells is synchronized within islets, and that the population of islets is also synchronized. We have recently developed a model in which pulsatile insulin secretion is produced as a result of calcium-driven electrical oscillations in combination with oscillations in glycolysis. We use this model to investigate possible mechanisms for intra islet and inter-islet synchronization. We show that electrical coupling is sufficient to synchronize both electrical bursting activity and metabolic oscillations. We also demonstrate that islets can synchronize by mutually entraining each other by their effects on a simple model "liver," which responds to the level of insulin secretion by adjusting the blood glucose concentration in an appropriate way. Since all islets are exposed to the blood, the distributed islet-liver system can synchronize the individual islet insulin oscillations. Thus, we demonstrate how intra-islet and inter-islet synchronization of insulin oscillations may be achieved. PMID- 15834003 TI - Coherent anti-stokes Raman scattering imaging of axonal myelin in live spinal tissues. AB - We present a vibrational imaging study of axonal myelin under physiological conditions by laser-scanning coherent anti-Stokes Raman scattering (CARS) microscopy. We use spinal cord white matter strips that are isolated from guinea pigs and kept alive in oxygen bubbled Krebs' solution. Both forward- and epi detected CARS are used to probe the parallel axons in the spinal tissue with a high vibrational contrast. With the CARS signal from CH2 vibration, we have measured the ordering degree and the spectral profile of myelin lipids. Via comparison with the ordering degrees of lipids in myelin figures formed of controlled lipid composition, we show that the majority of the myelin membrane is in the liquid ordered phase. By measuring the myelin thickness and axon diameter, the value of g ratio is determined to be 0.68 with forward- and 0.63 with epi detected CARS. Detailed structures of the node of Ranvier and Schmidt-Lanterman incisure are resolved. We have also visualized the ordering of water molecules between adjacent bilayers inside the myelin. Our observations provide new insights into myelin organization, complementary to the knowledge from light and electron microscopy studies of fixed and dehydrated tissues. In addition, we have demonstrated simultaneous CARS imaging of myelin and two-photon excitation fluorescence imaging of intra- and extraaxonal Ca2+. The current work opens up a new approach to the study of spinal cord injury and demyelinating diseases. PMID- 15834004 TI - An empirical extremum principle for the hill coefficient in ligand-protein interactions showing negative cooperativity. AB - The Hill coefficient (nH) is a central parameter in the study of ligand-protein interactions, which measures the degree of cooperativity between subunits that bind the ligand in multisubunit proteins. The most common usage of nH is as an estimate of the minimal number of interacting binding sites in positively cooperating systems. In the present study, a statistical interpretation of nH for a generalized system of multiple identical binding sites is developed. This interpretation is then applied to the derivation of an empirical extremum principle for nH in negatively cooperating systems of identical binding sites, which can be used for the estimation of the minimal number of interacting sites in such systems. PMID- 15834005 TI - Disorder influence on linear dichroism analyses of smectic phases. AB - Linear dichroism, the unequal absorption of parallel and perpendicular linear polarized light, is often used to determine the anisotropic ordering of rodlike polymers in a smectic phase, such as helices in a lipid bilayer. It is a measure of two properties of the sample: 1), orientation of the chromophore transition dipole moment (TDM) and 2), disorder. Since it is the orientation of the chromophore TDM that is needed for high resolution structural studies, it is imperative to either deconvolve sample disorder, or at a minimum, estimate its effect upon the calculated TDM orientation. Herein, a rigorous analysis of the effects of disorder is undertaken based on the recently developed Gaussian disorder model implemented in linear dichroism data. The calculation of both the rod tilt and rotational pitch angles as a function of the disorder and dichroism, yield the following conclusions: Disorders smaller than 5 degrees have a vanishingly small effect on the calculated polymer orientation, whereas values smaller than 10 degrees have a negligible effect on the calculated parameters. Disorders larger than 10 degrees have an appreciable effect on the calculated orientational parameters and as such must be estimated before any structural characterization. Finally the theory is tested on the HIV vpu transmembrane domain, employing experimental mosaicity measurements from x-ray reflectivity rocking scans and linear dichroism. PMID- 15834006 TI - Specific and nonspecific hybridization of oligonucleotide probes on microarrays. AB - Gene expression analysis by means of microarrays is based on the sequence specific binding of RNA to DNA oligonucleotide probes and its measurement using fluorescent labels. The binding of RNA fragments involving sequences other than the intended target is problematic because it adds a chemical background to the signal, which is not related to the expression degree of the target gene. The article presents a molecular signature of specific and nonspecific hybridization with potential consequences for gene expression analysis. We analyzed the signal intensities of perfect match (PM) and mismatch (MM) probes of GeneChip microarrays to specify the effect of specific and nonspecific hybridization. We found that these events give rise to different relations between the PM and MM intensities as function of the middle base of the PM, namely a triplet-like (C > G approximately T > A > 0) and a duplet-like (C approximately T > 0 > G approximately A) pattern of the PM-MM log-intensity difference upon binding of specific and nonspecific RNA fragments, respectively. The systematic behavior of the intensity difference can be rationalized on the level of basepairings of DNA/RNA oligonucleotide duplexes in the middle of the probe sequence. Nonspecific binding is characterized by the reversal of the central Watson-Crick (WC) pairing for each PM/MM probe pair, whereas specific binding refers to the combination of a WC and a self-complementary (SC) pairing in PM and MM probes, respectively. The Gibbs free energy contribution of WC pairs to duplex stability is asymmetric for purines and pyrimidines of the PM and decreases according to C > G approximately T > A. SC pairings on the average only weakly contribute to duplex stability. The intensity of complementary MM introduces a systematic source of variation which decreases the precision of expression measures based on the MM intensities. PMID- 15834007 TI - Unique translation initiation at the second AUG codon determines mitochondrial localization of the phage-type RNA polymerases in the moss Physcomitrella patens. AB - The nuclear genome of the moss Physcomitrella patens contains two genes encoding phage-type RNA polymerases (PpRPOT1 and PpRPOT2). Each of the PpRPOT1 and PpRPOT2 transcripts possesses two in-frame AUG codons at the 5' terminus that could act as a translational initiation site. Observation of transient and stable Physcomitrella transformants expressing the 5' terminus of each PpRPOT cDNA fused with the green fluorescent protein gene suggested that both PpRPOT1 and PpRPOT2 are not translated from the first (upstream) AUG codon in the natural context but translated from the second (downstream) one, and that these enzymes are targeted only to mitochondria, although they are potentially targeted to plastids when translation is forced to start from the first AUG codon. The influence of the 5' upstream sequence on the translation efficiency of the two AUG codons in PpRPOT1 and PpRPOT2 was quantitatively assessed using a beta-glucuronidase reporter. The results further supported that the second AUG codon is the sole translation initiation site in Physcomitrella cells. An Arabidopsis (Arabidopsis thaliana) RPOT homolog AtRpoT;2 that possesses two initiation AUG codons in its transcripts, as do the RPOTs of P. patens, has been regarded as a dually targeted protein. When the localization of AtRpoT;2 was tested using green fluorescent protein in a similar way, AtRpoT;2 was also observed only in mitochondria in many Arabidopsis tissues. These results suggest that, despite the presence of two in frame AUGs at the 5' termini of RPOTs in Physcomitrella and Arabidopsis, the second AUG is specifically recognized as the initiation site in these organisms, resulting in expression of a protein that is targeted to mitochondria. This finding may change the current framework of thinking about the transcription machinery of plastids in land plants. PMID- 15834008 TI - Arabidopsis CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting growth. AB - Rice (Oryza sativa), a monocotyledonous plant that does not cold acclimate, has evolved differently from Arabidopsis (Arabidopsis thaliana), which cold acclimates. To understand the stress response of rice in comparison with that of Arabidopsis, we developed transgenic rice plants that constitutively expressed CBF3/DREB1A (CBF3) and ABF3, Arabidopsis genes that function in abscisic acid independent and abscisic acid-dependent stress-response pathways, respectively. CBF3 in transgenic rice elevated tolerance to drought and high salinity, and produced relatively low levels of tolerance to low-temperature exposure. These data were in direct contrast to CBF3 in Arabidopsis, which is known to function primarily to enhance freezing tolerance. ABF3 in transgenic rice increased tolerance to drought stress alone. By using the 60 K Rice Whole Genome Microarray and RNA gel-blot analyses, we identified 12 and 7 target genes that were activated in transgenic rice plants by CBF3 and ABF3, respectively, which appear to render the corresponding plants acclimated for stress conditions. The target genes together with 13 and 27 additional genes are induced further upon exposure to drought stress, consequently making the transgenic plants more tolerant to stress conditions. Interestingly, our transgenic plants exhibited neither growth inhibition nor visible phenotypic alterations despite constitutive expression of the CBF3 or ABF3, unlike the results previously obtained from Arabidopsis where transgenic plants were stunted. PMID- 15834009 TI - Citrate secretion coupled with the modulation of soybean root tip under aluminum stress. Up-regulation of transcription, translation, and threonine-oriented phosphorylation of plasma membrane H+-ATPase. AB - The aluminum (Al)-induced secretion of citrate has been regarded as an important mechanism for Al resistance in soybean (Glycine max). However, the mechanism of how Al induces citrate secretion remains unclear. In this study, we investigated the regulatory role of plasma membrane H+-ATPase on the Al-induced secretion of citrate from soybean roots. Experiments performed with plants grown in full nutrient solution showed that Al-induced activity of plasma membrane H+-ATPase paralleled secretion of citrate. Vanadate and fusicoccin, an inhibitor and an activator, respectively, of plasma membrane H+-ATPase, exerted inhibitory and stimulatory effects on the Al-induced secretion of citrate. Higher activity of plasma membrane H+-ATPase coincided with more citrate secretion in Al-resistant than Al-sensitive soybean cultivars. These results suggested that the effects of Al stress on citrate secretion were mediated via modulation of the activity of plasma membrane H+-ATPase. The relationship between the Al-induced secretion of citrate and the activity of plasma membrane H+-ATPase was further demonstrated by analysis of plasma membrane H+-ATPase transgenic Arabidopsis (Arabidopsis thaliana). When plants were grown on Murashige and Skoog medium containing 30 microM Al (9.1 microM Al3+ activity), transgenic plants exuded more citrate compared with wild-type Arabidopsis. Results from real-time reverse transcription PCR and immunodetection analysis indicated that the increase of plasma membrane H+-ATPase activity by Al is caused by transcriptional and translational regulation. Furthermore, plasma membrane H+-ATPase activity and expression were higher in an Al-resistant cultivar than in an Al-sensitive cultivar. Al activated the threonine-oriented phosphorylation of plasma membrane H+-ATPase in a dose- and time-dependent manner. Taken together, our results demonstrated that up regulation of plasma membrane H+-ATPase activity was associated with the secretion of citrate from soybean roots. PMID- 15834010 TI - Ethylene insensitivity conferred by the Green-ripe and Never-ripe 2 ripening mutants of tomato. AB - The ripening of a fleshy fruit represents the summation of an array of biochemical processes that are regulated by interactions between developmental programs and environmental inputs. Analysis of tomato (Solanum lycopersicum) mutants and inhibitor studies indicate that ethylene is necessary for full development of the ripening program of climacteric fruit such as tomato, yet ethylene alone is not sufficient. This suggests that an interaction between ethylene and nonethylene (or developmental) pathways mediates ripening. In this study, we have examined the physiological basis for ripening inhibition of the dominant Green-ripe (Gr) and Never-ripe 2 (Nr-2) mutants of tomato. Our data suggest that this inhibition is due to ethylene insensitivity in mutant fruit. Further investigation of ethylene responses in Gr and Nr-2 plants also revealed weak ethylene insensitivity during floral senescence and abscission and, during inhibition of root elongation, a phenotype associated with the triple response. However, ethylene-induced inhibition of hypocotyl elongation and petiole epinasty are normal in Gr and Nr-2, suggesting that these loci regulate a subset of ethylene responses. We have mapped both dominant mutations to a 2-cM overlapping region of the long arm of chromosome 1 of tomato, a region not previously linked to any known ethylene signaling loci. The phenotypic similarity and overlapping map location of these mutations suggest Gr and Nr-2 may be allelic and may possibly encode a novel component of the ethylene response pathway. PMID- 15834011 TI - Characterization of arsenate reductase in the extract of roots and fronds of Chinese brake fern, an arsenic hyperaccumulator. AB - Root extracts from the arsenic (As) hyperaccumulating Chinese brake fern (Pteris vittata) were shown to be able to reduce arsenate to arsenite. An arsenate reductase (AR) in the fern showed a reaction mechanism similar to the previously reported Acr2p, an AR from yeast (Saccharomyces cerevisiae), using glutathione as the electron donor. Substrate specificity as well as sensitivity toward inhibitors for the fern AR (phosphate as a competitive inhibitor, arsenite as a noncompetitive inhibitor) was also similar to Acr2p. Kinetic analysis showed that the fern AR had a Michaelis constant value of 2.33 mM for arsenate, 15-fold lower than the purified Acr2p. The AR-specific activity of the fern roots treated with 2 mM arsenate for 9 d was at least 7 times higher than those of roots and shoots of plant species that are known not to tolerate arsenate. A T-DNA knockout mutant of Arabidopsis (Arabidopsis thaliana) with disruption in the putative Acr2 gene had no AR activity. We could not detect AR activity in shoots of the fern. These results indicate that (1) arsenite, the previously reported main storage form of As in the fern fronds, may come mainly from the reduction of arsenate in roots; and (2) AR plays an important role in the detoxification of As in the As hyperaccumulating fern. PMID- 15834012 TI - Systems rebalancing of metabolism in response to sulfur deprivation, as revealed by metabolome analysis of Arabidopsis plants. AB - Sulfur is an essential macro-element in plant and animal nutrition. Plants assimilate inorganic sulfate into two sulfur-containing amino acids, cysteine and methionine. Low supply of sulfate leads to decreased sulfur pools within plant tissues. As sulfur-related metabolites represent an integral part of plant metabolism with multiple interactions, sulfur deficiency stress induces a number of adaptive responses, which must be coordinated. To reveal the coordinating network of adaptations to sulfur deficiency, metabolite profiling of Arabidopsis has been undertaken. Gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry techniques revealed the response patterns of 6,023 peaks of nonredundant ion traces and relative concentration levels of 134 nonredundant compounds of known chemical structure. Here, we provide a catalogue of the detected metabolic changes and reconstruct the coordinating network of their mutual influences. The observed decrease in biomass, as well as in levels of proteins, chlorophylls, and total RNA, gives evidence for a general reduction of metabolic activity under conditions of depleted sulfur supply. This is achieved by a systemic adjustment of metabolism involving the major metabolic pathways. Sulfur/carbon/nitrogen are partitioned by accumulation of metabolites along the pathway O-acetylserine to serine to glycine, and are further channeled together with the nitrogen-rich compound glutamine into allantoin. Mutual influences between sulfur assimilation, nitrogen imbalance, lipid breakdown, purine metabolism, and enhanced photorespiration associated with sulfur deficiency stress are revealed in this study. These responses may be assembled into a global scheme of metabolic regulation induced by sulfur nutritional stress, which optimizes resources for seed production. PMID- 15834013 TI - What causes lacunar stroke? PMID- 15834014 TI - Imaging Huntington's disease (HD) brains--imagine HD trials! PMID- 15834015 TI - Weight bearing asymmetry in standing hemiparetic patients. PMID- 15834016 TI - Prognosis in the acute motor axonal form of Guillain-Barre syndrome. PMID- 15834018 TI - Self awareness and personality change in dementia. AB - BACKGROUND: Loss of insight is a core diagnostic criterion for frontotemporal dementia (FTD), whereas failure to recognise cognitive deficits and unawareness of disease (anosognosia) are well established findings in Alzheimer's disease (AD). However, self awareness of personality has not been quantified in these patient groups. METHODS: Twenty two patients (12 with frontal variant FTD; 10 with early AD) and 11 older adult normal controls completed self report questionnaires (the Interpersonal Adjectives Scales) describing their current personality. First degree relative informants completed two questionnaires, one describing the subject's current personality, the other retrospectively describing the subject's personality before disease onset. Differences between subject and informant reports of current personality were used to measure the accuracy of self awareness. RESULTS: Discriminant function analysis showed significant differences in self awareness among the three groups, with those in the FTD group showing the greatest magnitude of error in the largest number of personality dimensions (dominance, submissiveness, cold heartedness, introversion, and ingenuousness). Despite personality changes over time, patients with AD showed accurate self awareness in all personality dimensions except submissiveness and extraversion. Normal controls showed a pattern of underestimating positive qualities, whereas patients with FTD exaggerated positive qualities and minimised negative qualities. For the personality facets showing impaired insight, the self reports of patients with FTD and AD most closely matched their premorbid personalities, suggesting a failure to update their self image after disease onset. CONCLUSIONS: This study operationalises research criteria for loss of insight in FTD. PMID- 15834017 TI - Neurogenetics II: complex disorders. AB - The genetic analysis of common neurological disorders will be a difficult and protracted endeavour. Genetics is only one of many disciplines that will be required but it has already thrown considerable light on the aetiology of several major neurological disorders through the analysis of rare inherited subgroups. The identification of individual susceptibility genes with variants of smaller effect will be more difficult but there is no sharp demarcation between large and small genetic effects, so that many new and important insights will emerge using existing and new technologies. The availability of improved neuroimaging, better animal models of disease and new genetic tools, such as high-throughput gene chips, expression microarrays and proteomics, are extending the range of traditional genetic mapping tools. Finally, an understanding of the genetic and epigenetic mechanisms that restrain the differentiation and integration of human neural stem cells into mature neuronal networks could have a major impact on clinical practice. These approaches will be illustrated in the context of Alzheimer disease, Parkinson disease and synucleinopathies, tauopathies, amyotrophic lateral sclerosis and stroke. PMID- 15834019 TI - Rate of progression of cognitive decline in Alzheimer's disease: effect of butyrylcholinesterase K gene variation. AB - OBJECTIVE: To determine whether individuals with Alzheimer's disease (AD) and the K variant allele of butyrylcholinesterase have a slower rate of cognitive decline than those without the K variant allele of butyrylcholinesterase. METHOD: The cognitive status of 339 community based subjects with AD was assessed with the Mini Mental State Examination at baseline and yearly over a three year follow up period. The rates of cognitive decline of subjects with and without the K variant allele were compared. RESULT: Presence of the K allele was associated with a slower average rate of cognitive decline in subjects with severe AD. CONCLUSIONS: This finding is consistent with the suggestion that the K variant of butyrylcholinesterase has an important role in disease progression in AD, and this may have implications for treatment. PMID- 15834020 TI - A longitudinal study of sentence comprehension difficulty in primary progressive aphasia. AB - CONTEXT: Patients with primary progressive aphasia have sentence comprehension difficulty, but the longitudinal course of this deficit has not been investigated. OBJECTIVE: To determine how grammatical, single word meaning, and working memory factors contribute to longitudinal decline of sentence comprehension in primary progressive aphasia. We hypothesised partially distinct patterns of sentence comprehension difficulty in subgroups of patients with progressive non-fluent aphasia (PNFA) and semantic dementia (SD). DESIGN: Cohort. SETTING: Institutional out patient referral centre. PATIENTS: PNFA (n = 14), SD (n = 10). MAIN OUTCOME MEASURE: Sentence comprehension accuracy. RESULTS: PNFA patients were significantly impaired at understanding grammatically complex sentences when first seen, and this was more evident than impairment of their comprehension of grammatically simple sentences (p<0.05). Comprehension of grammatically complex sentences correlated with their working memory deficit at presentation (p<0.05). PNFA patients showed modest decline over time in grammatical comprehension. In SD, comprehension of grammatically complex sentences was not more impaired than comprehension of grammatically simple sentences when first seen, but these patients demonstrated a significant longitudinal decline in understanding grammatically complex sentences (p<0.05). Cox regression analyses showed that a deficit in single word meaning contributes to the progressive impairment for grammatically complex sentences in SD (p<0.05), but working memory does not contribute to longitudinal decline in PNFA. CONCLUSION: Patients with PNFA and SD have sentence comprehension difficulty, but distinct factors contribute to this impairment during the course of their disease. PMID- 15834021 TI - Progression of structural neuropathology in preclinical Huntington's disease: a tensor based morphometry study. AB - BACKGROUND AND OBJECTIVES: Regional cerebral atrophy occurs in carriers of the Huntington's disease (HD) gene mutation before clinical diagnosis is possible. The current inability to reliably measure progression of pathology in this preclinical phase impedes development of therapies to delay clinical onset. We hypothesised that longitudinal statistical imaging would detect progression of structural pathology in preclinical carriers of the HD gene mutation, in the absence of measurable clinical change. METHODS: Thirty subjects (17 preclinical mutation positive, 13 mutation negative) underwent serial clinical and magnetic resonance imaging (MRI) assessments over an interval of 2 years. Statistically significant changes in regional grey and white matter volume on MRI were analysed using tensor based morphometry (TBM). This technique derives a voxel-wise estimation of regional tissue volume change from the deformation field required to warp a subject's early to late T1 images. RESULTS: Over 2 years, there was progressive regional grey matter atrophy in mutation-positive relative to negative subjects, without significant clinical progression of disease. Significant grey matter volume loss was limited to bilateral putamen and globus pallidus externa (GPe), left caudate nucleus, and left ventral midbrain in the region of the substantia nigra. CONCLUSIONS: While these results are consistent with previous cross sectional pathologic and morphometric studies, significant progression of atrophy in HD before the onset of significant clinical decline is now demonstrable with longitudinal statistical imaging. Such measures could be used to assess the efficacy of potential disease modifying drugs in slowing the progression of pathology before confirmed clinical onset of HD. PMID- 15834023 TI - Robert Barany 1876-1936. PMID- 15834022 TI - The presence of glutamic acid at positions 71 or 74 in pocket 4 of the HLA DRbeta1 chain is associated with the clinical course of multiple sclerosis. AB - BACKGROUND: Primary progressive multiple sclerosis (PP-MS) differs from relapsing remitting or secondary progressive MS (RR/SP-MS) in ways suggesting differences in the pathogenic pathways. Susceptibility to both PP-MS and RR/SP-MS is linked to carriage of the HLA molecule DRB1*1501. Several serologically defined HLA-DR groups (DR1, DR4, DR6, and DR9) occur less often in RR/SP-MS than in controls. Some or all of the HLA-DR molecules encoded by alleles in these serologically defined groups have a negatively charged glutamic acid at residue 71 or 74 of the beta1 chain (beta1(71)/beta1(74)). Residues at these positions are important in the formation of pocket 4 in the antigen binding site of the HLA-DR molecule. OBJECTIVES: To investigate whether the presence of alleles encoding HLA-DR molecules containing glutamic acid at beta1(71)/beta1(74) correlates with the course of MS. METHODS: HLA-DR and HLA-DQ alleles and genotypes were analysed in 121 MS patients (50 with PP-MS) and 109 controls by molecular typing. RESULTS: Alleles encoding HLA-DR molecules containing a glutamic acid at beta1(71)/beta1(74) occurred less often in patients with RR/SP-MS than in those with PP-MS or controls. In subjects not carrying the DRB1*1501 allele, a much higher proportion of PP-MS patients carried alleles encoding HLA-DR molecules containing a glutamic acid at beta1(71)/beta1(74) than did RR/SP-MS patients or controls. CONCLUSIONS: The amino acid residues involved in determining the shape and charge of pocket 4 of the HLA-DR beta1 chain could influence the clinical course of MS by determining protection against RR/SP-MS or susceptibility to the development of PP-MS. PMID- 15834024 TI - Severe haemodynamic stress in selected subtypes of patients with moyamoya disease: a positron emission tomography study. AB - BACKGROUND: The optimum management of patients with moyamoya disease remains controversial. OBJECTIVES: To examine retrospectively the correlation between the degree of haemodynamic stress and the clinical presentation by measuring cerebral haemodynamics and metabolism using positron emission tomography (PET). METHODS: 57 patients with moyamoya disease (mean age 32 years, range 12 to 64), classified into five groups according to clinical manifestations, underwent PET measurement of cerebral blood flow (CBF), cerebral blood volume (CBV), cerebral metabolic rate for oxygen (CMR(O2)), and oxygen extraction fraction (OEF) using (15)O labelled gases. The regional values in patient groups were compared with a normal group. RESULTS: CBF in non-symptomatic patients, patients presenting with transient ischaemic attacks (TIA), and patients with haemorrhagic onset (H) was not significantly lower than in normal controls in any region. CBV in the TIA group and in patients with infarction associated with TIA (I/TIA) was significantly higher than in the controls in most regions. OEF in the frontal, parietal, and temporal cortex was significantly higher in the I/TIA group than in the controls. Patients in the H group and those with a permanent deficit with infarction (PD group) had decreased metabolism with normal OEF. Multivariate analysis to test the distribution of the three dimensional vector (CBF, CBV, OEF) showed significant differences between every possible pair among the six groups except NS v H and H v PD in the frontal cortex. CONCLUSIONS: The haemodynamic status of moyamoya disease is not uniform, and severe haemodynamic stress occurs in selected subgroups of patients. PMID- 15834025 TI - The vestibular control of balance after stroke. AB - OBJECTIVES: To examine vestibular control of balance in those who recovered the ability to stand after middle cerebral artery (MCA) stroke. METHODS: Sixteen patients with MCA stroke were compared with 10 age matched controls. Two additional patients were studied with isolated corticospinal tract lesions, one each at the level of the pons and medulla. Vestibular evoked postural responses were obtained using galvanic vestibular stimulation (GVS) while patients stood with their eyes closed and head facing forwards, equally loading both legs. The GVS response was characterised by measuring the amplitude of the stimulus evoked lateral forces acting through each leg and the lateral displacement of the axial skeleton. RESULTS: Lateral displacement and net lateral force following GVS were significantly larger after stroke. Unlike controls, the lateral forces in the stroke group were asymmetrical, being enhanced on the side of the non-paretic limb and small on the side of the paretic limb. The degree of GVS evoked asymmetry correlated with corticospinal damage assessed using transcranial magnetic stimulation. A similar asymmetrical response was seen in the patient with the pontine lesion but not the patient with the medullary lesion. CONCLUSIONS: MCA stroke may disrupt corticobulbar projections to brainstem output pathways involved in vestibular control of balance. These projections are either collaterals of the corticospinal tract or lie close to that tract and terminate in the pons/upper medulla. This hypothesis accounts for the association between corticospinal tract damage and GVS response asymmetry, and the lack of GVS evoked asymmetry with corticospinal lesions below the rostral medulla. PMID- 15834026 TI - Atrial fibrillation as a predictive factor for severe stroke and early death in 15,831 patients with acute ischaemic stroke. AB - BACKGROUND: Atrial fibrillation (AF) is a common arrhythmia and a major risk factor for stroke. AIMS: To assess whether AF in patients with acute ischaemic stroke is associated with severe stroke and early death. MATERIALS/METHODS: Patients with acute ischaemic stroke (15,831) who were registered in the Japan Multicenter Stroke Investigators' Collaboration registry were analysed. The AF group comprised 3335 (21.1%) patients (median age, 75 years) and the non-AF group comprised 12,496 (78.9%) patients (median age, 70 years). The association between AF and severe stroke and early death was investigated by means of multivariate logistic regression analysis. RESULTS: The admission National Institutes of Health Stroke Scale (NIHSS) score of the AF group was higher than that of the non AF group (median, 12 v 5; p < 0.0001). Multivariate logistic regression analyses found that female sex, advanced age, AF, and a history of stroke were independent factors associated with severe stroke (NIHSS score, > or = 11). The mortality rate within 28 days after admission was 11.3% in the AF group and 3.4% in the non AF group (p < 0.0001). Multivariate logistic regression analyses identified older age, AF, and NIHSS score at admission as independent factors associated with early death. CONCLUSION: AF was a predictive factor for severe stroke and early death in acute ischaemic stroke. Careful cardiac evaluation and appropriate treatment are needed to improve outcome in patients with acute stroke and AF. PMID- 15834027 TI - Bilateral thalamic deep brain stimulation: midline tremor control. AB - OBJECTIVES: To determine the efficacy of bilateral deep brain stimulation (DBS) for management of midline tremor (head, voice, tongue, trunk) in patients with essential tremor. DESIGN: Prospective assessment of tremor at baseline (presurgical), and postoperatively at 1, 3, and 12 months, and annually thereafter. METHODS: A clinical series of 22 individuals undergoing staged, bilateral DBS for treatment of essential tremor. The tremor rating scale was the primary outcome measure. RESULTS: Midline tremor showed significant improvement with stimulation "on" at nearly every postoperative interval when compared with stimulation "off" and with baseline tremor. Bilateral stimulation was associated with a significant incremental improvement in midline tremor control compared with unilateral stimulation: average "stimulation on" percentage change in midline tremor from the unilateral to bilateral period was 81%. Head and voice tremor showed the most consistent improvement. Among those requiring a change in stimulation parameters because of side effects, dysarthria, disequilibrium, motor disturbances, and paraesthesiae were the most common. Dysarthria was more common with bilateral (n = 6; 27%) than with unilateral (n = 0) stimulation. Stimulation parameters remained largely unchanged after the first three months. Nine of 44 leads placed (20%) required subsequent repositioning or replacement. CONCLUSIONS: Unilateral thalamic stimulation significantly improves midline tremor, and subsequent bilateral thalamic stimulation offers an additional incremental improvement in midline tremor control. PMID- 15834028 TI - Predictors of good outcome in medium to large spontaneous supratentorial intracerebral haemorrhages. AB - OBJECTIVE: To determine potential predictors of good outcome in primary medium to large intracerebral haemorrhages (ICH) which could be useful for selecting patients for surgical procedures. METHODS: Subjects were 138 patients with spontaneous hemispheric ICH >20 ml. They were non-surgically treated and were admitted consecutively to 15 hospitals within the first 12 hours of symptom onset (mean (SD), 5.8 (3.1) hours). Haematoma volume was measured on computed tomography (CT) at admission. Stroke severity was assessed by the Canadian stroke scale (CSS). Good outcome was defined as modified Rankin score < or =2 at three months. RESULTS: At the end of the follow up period, 45 patients (32.6%) had good outcome. Baseline stroke severity, systolic and diastolic blood pressure, body temperature, and acute phase reaction biochemical markers (ESR, C-reactive protein, fibrinogen, neutrophil count) were significantly associated with good outcome in bivariate analyses. Of the initial CT scan variables, intraventricular contamination, deep location, mass effect, and greater ICH volume were related to poor outcome. On multiple logistic regression analysis, cortical location of bleeding (odds ratio 3.79 (95% confidence interval 1.2 to 12.01); p = 0.023), high CSS score (OR 2.3 (1.6 to 3.1); p<0.0001), and low fibrinogen concentrations (OR 0.92 (0.87 to 0.97); p = 0.001) were independent predictors of good outcome. These three factors correctly classified 85% of patients. CONCLUSIONS: Good outcome in medium to large ICH can be predicted on admission by three readily assessable factors (CSS score, ICH location, and fibrinogen levels). These predictors may be helpful in selecting patients for surgical treatment. PMID- 15834029 TI - Association of a polymorphism of the transforming growth factor-beta1 gene with cerebral amyloid angiopathy. AB - BACKGROUND: A recent study showed that transforming growth factor-beta1 (TGF beta1) induces amyloid-beta deposition in cerebral blood vessels and meninges of a transgenic mouse model of Alzheimer's disease (AD), and that TGF-beta1 mRNA levels are correlated with cerebral amyloid angiopathy (CAA) in human AD brains. A T/C polymorphism at codon 10 in exon 1 of the TGF-beta1 gene has been reported to be associated with the serum TGF-beta1 concentration. We investigated whether the TGF-beta1 polymorphism is associated with the risk of CAA. METHODS: The association between the severity of CAA and the T/C polymorphism at codon 10 in exon 1 of the TGF-beta1 was investigated in 167 elderly Japanese autopsy cases, including 73 patients with AD. The apolipoprotein E (APOE) genotype was also determined. RESULTS: The genotypes (TT/ TC/ CC) were associated with the severity of CAA significantly in all patients (p = 0.0026), in non-AD patients (p = 0.011), and APOE non-epsilon4 carriers (p = 0.0099), but not in AD patients or APOE epsilon4 carriers. The number of the T alleles positively correlated with the severity of CAA in all patients (p = 0.0011), non-AD patients (p = 0.0026), and APOE non-epsilon4 carriers (p = 0.0028), but not in AD patients or APOE epsilon4 carriers. The polymorphism was not significantly associated with AD. CONCLUSIONS: Our results suggest that the polymorphism in TGF-beta1 is associated with the severity of CAA, especially in non-AD patients and APOE non-epsilon4 carriers. PMID- 15834030 TI - A double blind, randomised study of sildenafil citrate for erectile dysfunction in men with multiple sclerosis. AB - OBJECTIVE: Identifying and effectively treating erectile dysfunction (ED) can result in an improvement of the quality of life (QoL) in men with multiple sclerosis (MS). METHODS: This randomised, double blind (DB), placebo controlled, flexible dose study with an open label extension (OLE) assessed efficacy, QoL, and safety of sildenafil citrate in men with MS and ED. Overall, 217 men received sildenafil (25-100 mg; n = 104) or placebo (n = 113) for 12 weeks. Efficacy was assessed by the International Index of Erectile Function (IIEF) questionnaire that includes questions on achieving (Q3) and maintaining (Q4) an erection as well as a global efficacy question (GEQ). QoL was also assessed. RESULTS: After 12 weeks, patients receiving sildenafil had higher mean scores for IIEF Q3 and Q4 compared with those receiving placebo (p<0.0001), and 89% (92/103) reported improved erections compared with 24% (27/112) of patients receiving placebo (p<0.0001). At the end of the OLE phase, 95% of men reported improved erections. Patients receiving placebo during the DB phase showed a nearly fourfold increase in improved erections (97% v 26%). Men receiving sildenafil also showed improvements in five of the eight general QoL questions compared with men receiving placebo (p<0.05). The total mean score for the QoL questionnaire improved by 43% for the sildenafil group versus 13% for the placebo group (p<0.0001). Treatment related AEs were predominantly mild in nature, and no patient discontinued due to an AE. CONCLUSION: Sildenafil treatment for ED in men with MS was effective and well tolerated, and resulted in significant improvements in both general and disease specific QoL variables. PMID- 15834031 TI - Homocysteine and related genetic polymorphisms in Down's syndrome IQ. AB - OBJECTIVE: Down's syndrome (DS) is the most frequent genetic cause of Alzheimer type dementia. Its metabolic phenotype involves an increased trans-sulphuration of homocysteine. The aim of the present study was to investigate the influence of homocysteinaemia (t-Hcys), folate, vitamin B(12), and related polymorphisms on intelligence quotient (IQ) in DS. METHODS: The IQ of 131 patients with trisomy 21 from a specialist centre in Sicily was determined and classified according to DMS IV. The effects of age, folate, vitamin B(12), t-Hcys, and genetic polymorphisms on IQ were evaluated separately and in combination using regression analyses. RESULTS: IQ was significantly lower in DS patients with t-Hcys >7.5 micromol/l (median) and in those who were carriers of methylenetetrahydrofolate reductase (MTHFR) 677 T allele and of methylenetetrahydrofolate reductase 677 T and transcobalamin 776 G combined alleles (p = 0.0013, p = 0.0165, and p = 0.0074, respectively). The IQ correlated significantly with t-Hcys and folate in single and multiple regression analyses, independently of age. In addition, t-Hcys >9.6 micromol/l (upper quartile) was found to be associated with low IQ (<40, median of study group) with an odds ratio of 2.61 (p = 0.0203). The odds ratio was increased by threefold in carriers of MTHFR 677T allele. The MTHFR 677T allele/transcobalamin 776 G allele combination was associated with the risk of DS patients to have an IQ less that the median with an odds ratio of 2.68 (95% CI 1.26 to 5.70, p = 0.0104). CONCLUSION: This study found evidence of an association between t-Hcys and MTHFR 677 T and transcobalamin 776 G alleles with IQ in patients with DS. The association may be related to a defective remethylation of homocysteine, affecting IQ. PMID- 15834032 TI - Seizure outcome after epilepsy surgery in patients with normal preoperative MRI. AB - OBJECTIVE: To determine outcome after epilepsy surgery in patients with normal preoperative magnetic resonance imaging (MRI). METHODS: 24 adult and paediatric patients with normal preoperative MRIs were studied. They underwent epilepsy surgery between 1994 and 2001 and had at least one year of follow up. RESULTS: At the most recent follow up, nine patients (37%) were seizure-free and 18 (75%) had at least a 90% reduction in seizure frequency with weekly or monthly seizures. Seizure freedom was not significantly different after resections in frontal (5/9) or temporal regions (4/13) (p = 0.24, Fisher's exact test), or among patients with or without localising features on EEG, PET, or ictal SPECT. Subdural grids, used in 15 of 24 patients, helped tailor resections but were not associated with differences in outcome. Histopathology showed cortical dysplasia in 10 patients (42%), non-specific findings in 13 (54%), and hippocampal sclerosis in one (4%). Cortical dysplasia was seen in seven patients with frontal resection (78%) and non-specific findings in nine (69%) with temporal resection. Seizure outcome did not differ on the basis of location of resection or histopathology. CONCLUSIONS: While these results were less favourable than expected for patients with focal epileptogenic lesions seen on MRI, they represented worthwhile improvement for this patient population with high preoperative seizure burden. In this highly selected group, no single test or combination of tests further predicted postoperative seizure outcome. PMID- 15834033 TI - Patterns and severity of neuromuscular transmission failure in seronegative myasthenia gravis. AB - OBJECTIVES: To compare the clinical and electrophysiological features of myasthenia gravis (MG) patients with (seropositive) or without (seronegative) antibodies to acetylcholine receptor. To investigate whether antibodies to muscle specific kinase (MuSK) and ryanodine receptor (RyR) are associated with particular features. METHODS: Clinical profiles and single fibre electromyography (SFEMG) in the extensor digitorum communis (EDC) were reviewed in consecutive 57 seropositive and 13 seronegative patients. Antibodies to MuSK and RyR were measured by immunoassays. RESULTS: Of the 13 seronegative patients, four (31%) were positive for MuSK antibodies and seven (54%) were positive for RyR antibodies, including all four MuSK positive patients. Clinical features were similar at presentation for seropositive and seronegative patients, but MuSK positive patients frequently developed myasthenic crises. Despite the similar clinical severities at the time of examination, the proportion with positive jitter (93% of seropositive patients, 50% of MuSK positive patients, and 44% of MuSK negative patients) and the extent of jitter (mean consecutive difference: 76 micros in seropositive patients, 36 micros in MuSK positive patients, and 30 micros in MuSK negative patients) were less in seronegative MG patients compared with seropositive MG patients. CONCLUSIONS: Seronegative MG is heterogeneous with respect to the presence of antibodies to MuSK. Impairment of neuromuscular synaptic transmission in EDC is less marked in seronegative than seropositive MG despite the similar clinical severity. This discrepancy may partly reflect the distribution of affected muscles in seronegative patients, but it is possible that other factors, such as impaired excitation-contraction coupling resulting from RyR antibodies, contribute to the clinical phenotype. PMID- 15834034 TI - Recovery patterns and long term prognosis for axonal Guillain-Barre syndrome. AB - BACKGROUND: Little is known about the long term prognosis for patients the severe acute motor axonal neuropathy (AMAN) form of Guillain-Barre syndrome (GBS), unlike those with acute inflammatory demyelinating neuropathy (AIDP). OBJECTIVE: To clarify the long term prognosis for patients with AMAN. METHODS: Clinical recovery and outcome in 97 consecutive GBS patients were reviewed. RESULTS: Electrodiagnostic criteria showed that 44 patients (45%) had AMAN, 33 (34%) had AIDP, and 20 (21%) were unclassified. Most of the severely affected patients had received plasmapheresis or immunoglobulin therapy. Slow recovery (inability to walk independently at six months after onset) was found in six of the AMAN patients (14%) and in two of the AIDP patients (6%). Of the six AMAN patients, four could walk independently one year after the onset, and the other two could walk independently at 28 and 57 months after onset. Of the two AIDP patients, one could walk at nine months after the onset while the other died of pneumonia seven months after onset. CONCLUSIONS: AMAN electrodiagnosis is not always a marker of poor recovery. Almost all the severe AMAN patients who had slow recoveries over the first six months could eventually walk independently, although some required several years. PMID- 15834035 TI - Evaluating neurorehabilitation: lessons from routine data collection. AB - BACKGROUND: Clinical databases are being used increasingly to assess outcomes in healthcare services to provide evidence of clinical effectiveness in routine clinical practice. OBJECTIVES: To explore the benefits of a database for routine collection of clinical outcomes within an inpatient neurorehabilitation setting; determine the effectiveness of inpatient neurorehabilitation in a range of neurological conditions; and determine variables influencing change in functional outcome. METHODS: Over a nine year period, demographic and diagnostic characteristics were collected for the 1458 patients admitted consecutively to a neurorehabilitation unit. The level of function was measured on admission and discharge using the Barthel Index (BI) and Functional Independence Measure (FIM). Patient perception of rehabilitation benefit was evaluated using visual analogue scales (VAS). RESULTS: Of the 1413 patients (mean (SD) age 48 (14.8), range 16 to 87) whose length of stay was more than 10 days (mean 34 (24) range 10 to 184), 282 had stroke, 614 multiple sclerosis, 248 spinal cord injuries, 93 a neuromuscular condition, and 176 other brain pathology. Patients improved in functional ability as measured by both BI and the FIM motor subscale (effect sizes 0.93 to 1.44 and 1.01 to 1.48, respectively). VAS ratings demonstrated high levels of patient perceived benefit. Diagnosis, functional activity score on admission, and length of stay were significant predictors of functional gain, explaining 44% of the variability in the change scores. CONCLUSIONS: Systematic collection, analysis, and interpretation of standardised clinical outcomes data are feasible within routine clinical practice, and provide evidence that inpatient rehabilitation is effective in improving functional level in neurologically impaired patients. These data complement those of clinical trials and are useful in informing and developing clinical and research practice. PMID- 15834036 TI - Lower urinary tract function in dementia of Lewy body type. AB - OBJECTIVE: Dementia of Lewy body (DLB) type is the second commonest degenerative cause of dementia and autonomic dysfunction has been recognised in DLB. Lower urinary tract (LUT) function in DLB has not been fully delineated. We investigated LUT function in DLB by evaluating clinical and urodynamic data. METHODS: We examined 11 patients (eight men, three women; age range 65-81; disease duration 2-14 years) with probable DLB. Urodynamic studies consisted of: measurement of postvoid residual in all patients, uroflowmetry in five, and electromyography (EMG) cystometry in seven. RESULTS: All patients had symptoms of LUT: urinary incontinence (urgency type/functional type due to dementia and immobility/both urgency and stress type in 7/2/1 patients, respectively); night time frequency; urgency; and daytime frequency and voiding difficulty. Seven had postvoid residuals, and three had residual urine volume >100 ml. Decreased urinary flow was seen in all five and detrusor overactivity in 5/7 patients who underwent flowmetry and EMG cystometry, respectively. Low compliance detrusor (storage phase, n = 2; with bethanechol supersensitivity), an underactive detrusor (n = 4), an acontractile detrusor (n = 1), and detrusor-sphincter dyssynergia (voiding phase) (n = 1) were also seen; 2/3 patients who underwent motor unit potential analysis had neurogenic changes. CONCLUSION: LUT dysfunction is a common feature in DLB, not only due to dementia and immobility, but also to central and peripheral types of somato-autonomic dysfunction. PMID- 15834037 TI - How to improve repetition ability in patients with Wernicke's aphasia: the effect of a disguised task. AB - Dissociation "automatico-voluntaire" is a symptom observed in aphasic patients. We elucidated the difference between voluntary and involuntary speech output in a quantitative manner using the same task materials in nine patients with Wernicke's aphasia. All the patients exhibited better ability and less paraphasias in a repetition task elicited in a disguised condition than in an ordinary repetition condition. This result indicates that the output difficulty in Wernicke's aphasia might be a disability of volitional control over the language system. PMID- 15834038 TI - Platysma sign in high cervical lesion. PMID- 15834039 TI - The spectrum of Notch3 mutations in 28 Italian CADASIL families. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is a cause of hereditary cerebrovascular disease. It results from mutations in the Notch3 gene, a large gene with 33 exons. A cluster of mutations around exons 3 and 4 was originally reported and limited scanning of these exons was suggested for the diagnosis in most cases. OBJECTIVE: To report Notch3 mutation analysis in 28 unrelated Italian CADASIL families from central and south Italy. RESULTS: The highest rate of mutations was found in exon 11 (21%) and only 18% of mutations were in exon 4. This may be related to the peculiar distribution of Notch3 mutations in the regions of origin of the families. CONCLUSIONS: The results suggest that limited scanning of exons 3 and 4 is inadvisable in CADASIL cases of Italian origin. PMID- 15834040 TI - The prevalence of cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) in the west of Scotland. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is caused by mutations of the Notch3 gene on 19p13. Varying phenotypic expression leads to under recognition and misdiagnosis. Prevalence therefore remains uncertain. We sought to estimate the prevalence of CADASIL in the west of Scotland. METHODS: A register for CADASIL was established at a regional neurosciences centre in 2002. All patients with genetically (exons 3, 4, 5, and 6) or histologically confirmed CADASIL residing in two defined administrative health areas were identified. Pedigree members at varying risk of carrying the mutation were also identified and the number of probable Notch3 mutation carriers in the defined population was predicted. Prevalence was calculated for definite CADASIL cases, with and without probable carrier numbers, based upon adult population figures from the 2002 national census. RESULTS: Twenty two individuals from seven pedigrees with confirmed CADASIL and resident in the defined geographical area were identified, yielding a prevalence of 1.98 (95% confidence interval 1.24-3.00) per 100 000 adults. An additional 37 individuals were predicted to be carriers of the Notch3 mutation, yielding a probable mutation prevalence of 4.14 (3.04-5.53) per 100,000 adults. CONCLUSIONS: The prevalence of genetically proven CADASIL was 1.98 per 100,000 adults in the defined population. This figure underestimates disease burden. PMID- 15834041 TI - Bilateral paramedian midbrain infarct: an uncommon variant of the "top of the basilar" syndrome. AB - Occlusion of the rostral portion of the basilar artery can result in ischaemia of the midbrain and thalami, as well as of the temporal and occipital lobes. The so called "top of the basilar" syndrome manifests clinically as numerous combinations of abnormalities of alertness, sleep-wake cycle, and behaviour and oculomotor or pupillomotor functions. A 67 year old man presented with bilateral internuclear ophthalmoplegia, rubral tremor, and daytime somnolence. He was awake during the night and mostly agitated and aggressive. An ischaemic lesion was visible on the magnetic resonance (MR) image in the central portion of the midbrain just ventral to the aqueduct, clearly affecting the paramedian structures bilaterally. MR angiographic images demonstrated a hypoplastic basilar artery ending in both superior cerebellar arteries. Both posterior cerebral arteries were seen to arise from the corresponding internal carotid arteries via the posterior communicating branches. This unique case of an acute bilateral paramedian infarct represents a highly uncommon variant of the "top of the basilar" syndrome and was due to the affected ischaemic territory--that is, the "distal field" of the variant basilar artery. PMID- 15834042 TI - Cognitive patterns and progression in multiple sclerosis: construction and validation of percentile curves. AB - BACKGROUND AND OBJECTIVES: Rate and pattern of progression of cognitive decline in multiple sclerosis (MS) has not been clearly identified. The present study aimed to identify correlations between cognitive tests and disease duration, construct longitudinal cognitive curves, and assess pattern of change over time. METHODS: The Neuropsychological Screening Battery for Multiple Sclerosis was administered in 150 consecutive MS patients, and tests that correlated with disease duration were identified. Percentile curves were constructed and the pattern of cognitive decline over time explored. The cognitive curves were validated in an additional group of 83 patients with MS. RESULTS: Three of four measures of the spatial recall test (SPART 7/24), and the paced auditory serial addition task for two seconds (PASAT 2'), correlated with disease duration. These tests were used to construct cross-sectional curves identifying the pattern of cognitive decline over time in the MS population. On the basis of this cross sectional analysis, the earliest cognitive decline occurred in the SPART 7/24 trials 1-5 between one and three years from onset, followed by decline in the SPART delayed recall between three and seven years, and then by decline in the PASAT 2' after seven years from onset. CONCLUSIONS: Verbal fluency and verbal memory appear to be affected earliest in MS. The pattern of cognitive decline is further characterised by a decrease in visuospatial learning, followed by delayed recall, and then by attention and information processing speed. Cognitive percentile curves can be used to evaluate the pattern of progression and identify patients at increased risk. PMID- 15834043 TI - Copper deficiency myeloneuropathy and pancytopenia secondary to overuse of zinc supplementation. AB - The haematological complications of acquired copper deficiency have been well documented, but the neurological complications have only recently been reported. An illustrative case of copper deficiency myeloneuropathy with pancytopenia is presented and the potential aetiologies and neurological manifestations of this deficiency state discussed. PMID- 15834044 TI - Phenotypic variability in a Chinese family with rimmed vacuolar distal myopathy. AB - BACKGROUND: UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) gene mutations have been found in patients with distal myopathy with rimmed vacuoles (DMRV). It is not clear how the same GNE gene mutations can result in different phenotypes in the same family with DMRV. METHODS: The clinical, neurophysiological, histopathological, and genetic characteristics of two patients with DMRV from a Chinese family from Taiwan were investigated. RESULTS: Two novel compound heterozygous mutations in different domains of the protein, Ile241Ser in the epimerase and Trp513stop in the kinase domain, were detected in both patients. However, the two patients demonstrated different patterns of disease progression: one had slow disease progression with a typical feature of DMRV (that is, weakness beginning in the distal leg muscles, typically anterior tibialis, with the quadriceps remaining relatively unaffected), and the other had rapid disease progression with an atypical presentation of DMRV. CONCLUSIONS: The results of the present study indicate that GNE gene mutations and probably modifier gene(s) or additional factors may result in different phenotypes of DMRV. PMID- 15834045 TI - A case with a 120 base pair insertional mutation in the prion protein gene: the first case in Japan. PMID- 15834046 TI - Identification of DNA markers for a transmissible Pseudomonas aeruginosa cystic fibrosis strain. AB - A number of transmissible Pseudomonas aeruginosa strains have been identified which potentially constitute an emerging threat to patients with cystic fibrosis (CF). We sought to identify DNA markers that were specific to a transmissible P. aeruginosa CF clone and evaluate these probes on a large collection of genotypically distinct P. aeruginosa strains. Using subtractive DNA hybridization, in combination with analysis using the P. aeruginosa PAO1 genome chip, DNA markers specific for or absent from the Manchester transmissible CF strain (MA) were identified. Five subtractive DNA hybridization markers (MA15, MA18, MA21, MA22, and MA30) were found to be specific to strain MA and were located within a novel 13,318-bp genomic island, designated the MA island. The MA island encoded 18 genes and consisted of two bacteriophage-like regions; one region encoded the MA-specific subtractive hybridization markers, while the other bacteriophage-like region contained a Vibrio cholera-like toxin gene. Probes MA15, MA18, MA21, MA22, and MA30 were all found to be specific to strain MA when a collection of 141 P. aeruginosa strains was examined by hybridization with each DNA marker. In contrast, a previously isolated DNA marker for the Liverpool transmissible CF strain, PS21, was not found to be specific, detecting two additional strain types in the collection screened. Both the Manchester and Liverpool strain types were not encountered in CF populations outside the United Kingdom. The MA genomic island and Vibrio cholera-like toxin gene within it constitute novel genetic factors associated with a transmissible P. aeruginosa strain and their role in pathogenesis remains to be determined. PMID- 15834047 TI - Chronic aeroallergen during infancy enhances eotaxin-3 expression in airway epithelium and nerves. AB - We have documented that exposure of rhesus monkeys to house dust mite aeroallergen during postnatal development resulted in significant recruitment of eosinophils into the airway mucosa (Clin Exp Allergy 33:1686-1694, 2003). Because eosinophils were not uniformly distributed throughout the five conducting airway generations examined, we speculated that trafficking within anatomic microenvironments of the lung is mediated by differential chemokine expression. To address this question, we used quantitative real-time RT-PCR to evaluate the related eosinophilic chemokines, eotaxin (CCL11), eotaxin-2 (CCL24), and eotaxin 3 (CCL26) within isolated airways of infant monkey lung. Overall, chemokine mRNA expression levels in house dust mite-exposed airways were as follows: eotaxin-3 > eotaxin > eotaxin-2. Immunofluorescence staining for eotaxin-3 and CC chemokine receptor 3 (CCR3) showed positive cells within epithelium and peripherally located nerve fiber bundles of the airway wall. Epithelial volume of eotaxin-3 within the trachea correlated with epithelial volume of major basic protein. CCR3+ and MHC Class II+ dendritic cells, but not eosinophils or mast cells, co localized within eotaxin-3+ nerve fiber bundles. We conclude that localized expression of eotaxin-3 plays an important role in the recruitment of diverse CCR3+ cell populations to different anatomic microenvironments within the infant airway in response to chronic allergen exposure. PMID- 15834052 TI - Risk of lymphoma in patients with RA treated with anti-TNFalpha agents. PMID- 15834053 TI - How to assess the impact of arthritis on the individual patient: the WHO ICF. AB - The ICF is not only a comprehensive and adequate framework for assessing the impact of arthritis on individual patients but also its impact on populations. The ICF framework and applications such as the ICF Core Sets for rheumatoid arthritis, osteoarthritis, osteoporosis, and low back pain are therefore likely to be used extensively not only in clinical practice but also in outcomes and rehabilitation research, education, health statistics, and regulation. PMID- 15834054 TI - MMP protein and activity levels in synovial fluid from patients with joint injury, inflammatory arthritis, and osteoarthritis. AB - OBJECTIVE: To determine protein and activity levels of matrix metalloproteinases 1 and 3 (MMP-1 and MMP-3) in synovial fluid of patients with knee joint injury, primary osteoarthritis, and acute pyrophosphate arthritis (pseudogout). METHODS: Measurements were done on knee synovial fluid obtained in a cross sectional study of cases of injury (n = 283), osteoarthritis (n = 105), and pseudogout (n = 65), and in healthy controls (n = 35). Activity of MMP-1 and MMP-3 in alpha(2) macroglobulin complexes was measured using specific low molecular weight fluorogenic substrates. ProMMP-1, proMMP-3, and TIMP-1 (tissue inhibitor of metalloproteinase 1) were quantified by immunoassay. RESULTS: Mean levels of proMMP-1, proMMP-3, and TIMP-1 were increased in injury, osteoarthritis, and pseudogout compared with controls. MMP-1 activity was increased in pseudogout and injury groups over control levels, whereas MMP-3 activity was increased only in the pseudogout group. The increase in MMP-1 activity coincided with a decrease in TIMP-1 levels in the injury group. CONCLUSIONS: Patients with joint injury have a persistent increase in proMMP-1 and proMMP-3 in synovial fluid and an increase in activated MMPs, which are not inhibited by TIMP. The differences in activation and inhibition patterns between the study groups are consistent with disease specific patterns of MMP activation and/or inhibition in joint pathology. PMID- 15834055 TI - Implication of interleukin 18 in production of matrix metalloproteinases in articular chondrocytes in arthritis: direct effect on chondrocytes may not be pivotal. AB - OBJECTIVE: To clarify the effect of interleukin (IL) 18 on cartilage degeneration by studying the profile of IL18 receptor (IL18R) on chondrocytes and the direct effect of IL18 on production of matrix metalloproteinases (MMPs), aggrecanases, and tissue inhibitors of metalloproteinases (TIMPs) in articular chondrocytes. METHODS: Monolayer cultured human articular chondrocytes were isolated from non arthritic subjects and patients with rheumatoid arthritis or osteoarthritis. Gene expression of IL18, IL18Ralpha, IL18Rbeta, MMPs, and aggrecanases was detected by RT-PCR. Protein levels of IL18Ralpha were analysed by flow cytometry. Protein levels of IL18, MMPs, and TIMPs were measured by ELISA. Aggrecanase-2 mRNA expression was quantitatively analysed by real time RT-PCR. Protein levels of signalling molecules were assayed by western blotting. RESULTS: IL18 mRNA was constitutively expressed in chondrocytes, and was enhanced by IL1beta stimulation. Flow cytometric analysis showed that IL1beta, tumour necrosis factor alpha, and IL18 up regulated IL18Ralpha expression levels. The level of IL18Rbeta mRNA was much lower than that of IL18Ralpha, and was slightly up regulated by IL1beta. In chondrocytes responding to IL18, IL18 (1-100 ng/ml) slightly increased the production of MMP-1, MMP-3, and MMP-13, which was blocked by NF kappaB inhibitor and p38 mitogen activated protein kinase inhibitor. IL18 up regulated mRNA expression of aggrecanase-2, but not aggrecanase-1. IL18 also slightly stimulated TIMP-1 production?through extracellular signal regulated kinase activation. CONCLUSION: IL18 induces production of MMPs from chondrocytes in inflammatory arthritis. Although the direct effect of IL18 on chondrocytes may not be pivotal for the induction of cartilage degeneration, IL18 seems to play some part in the degradation of articular cartilage in arthritis. PMID- 15834056 TI - Exercises versus arthroscopic decompression in patients with subacromial impingement: a randomised, controlled study in 90 cases with a one year follow up. AB - OBJECTIVES: To compare the effect of graded physiotherapeutic training of the rotator cuff versus arthroscopic subacromial decompression in patients with subacromial impingement. METHODS: Randomised controlled trial with 12 months' follow up in a hospital setting. Ninety consecutive patients aged 18 to 55 years were enrolled. Symptom duration was between six months and three years. All fulfilled a set of diagnostic criteria for rotator cuff disease, including a positive impingement sign. Patients were randomised either to arthroscopic subacromial decompression, or to physiotherapy with exercises aiming at strengthening the stabilisers and decompressors of the shoulder. Outcome was shoulder function as measured by the Constant score and a pain and dysfunction score. "Intention to treat" analysis was used, with comparison of means and control of confounding variables by general equation estimation analysis. RESULTS: Of 90 patients enrolled, 84 completed follow up (41 in the surgery group, 43 in the training group). The mean Constant score at baseline was 34.8 in the training group and 33.7 in the surgery group. After 12 months the mean scores improved to 57.0 and 52.7, respectively, the difference being non-significant. No group differences in mean pain and dysfunction score improvement were found. CONCLUSIONS: Surgical treatment of rotator cuff syndrome with subacromial impingement was not superior to physiotherapy with training. Further studies are needed to qualify treatment choice decisions, and it is recommended that samples are stratified according to disability level. PMID- 15834057 TI - Autoantibodies in biological agent naive patients with psoriatic arthritis. AB - BACKGROUND: Anti-tumour necrosis factor alpha (anti-TNFalpha) treatment may be associated with the production of autoantibodies, including lupus-specific autoantibodies. OBJECTIVE: To investigate the prevalence of autoantibodies in biological agent naive patients with psoriatic arthritis (PsA). METHODS: 94 consecutive, prospectively collected, biological agent naive patients with PsA at the University of Toronto PsA clinic underwent clinical and laboratory assessment. Disease activity was assessed by the number of actively inflamed joints, and the Psoriasis Activity and Severity Index (PASI) score. Antinuclear antibodies (ANA), rheumatoid factor (RF), double stranded DNA (dsDNA), Ro, La, Smith, and RNP were tested. Descriptive statistics and non-parametric tests were used to analyse the data. RESULTS: 44/94 (47%) patients with PsA were ANA positive (>/=1/40); 13/94 (14%) had a clinically significant titre of >/=1/80. Three per cent had dsDNA antibodies, 2% had RF and anti-Ro antibodies, 1% had anti-RNP antibodies, and none had anti-La or anti-Smith antibodies. CONCLUSIONS: The background prevalence of ANA >/=1/80 in patients with PsA was 14%, with very few patients having specific lupus antibodies. This should serve as a baseline figure for the frequency of autoantibodies in biological agent naive patients with PsA for studies of the use of anti-TNFalpha agents. PMID- 15834058 TI - White matter hyperintensities on brain magnetic resonance in systemic sclerosis. AB - OBJECTIVE: To evaluate the brain status of patients with systemic sclerosis (SSc). METHODS: Fourteen female patients with SSc aged 24-74, with a disease duration of 1-12 years and without other relevant systemic diseases, were enrolled. All patients and an age matched female control group (CG) of 14 clinically normal subjects, underwent brain magnetic resonance examination at 1.5 T; spin echo proton density weighted images were evaluated. Mann-Whitney U and Spearman rank correlation tests were used for statistical analysis. RESULTS: 170 white matter hyperintensities >/=2 mm in diameter were counted in the patient group (range 0-75, mean 12.1, median 4.5), only 13 in the CG (0-2, 0.9, 1, respectively), with a significant difference (p = 0.011). Moreover, 208 white matter hyperintensities <2 mm were found in the patient group (0-38, 14.9, 8, respectively), only 31 in the CG (0-7, 2.0, 1, respectively), with a significant difference (p = 0.006). No statistically significant correlation between the number of hyperintensities and either patient's age or disease duration was observed. CONCLUSION: White matter hyperintensities are more common in patients with SSc than in a CG. These findings might be related to obliterative microvascular processes due to the disease. Early brain involvement in patients with SSc may occur. PMID- 15834060 TI - Recurrent auricular chondritis and cartilage repair. PMID- 15834059 TI - Human parvovirus B19, varicella zoster virus, and human herpes virus 6 in temporal artery biopsy specimens of patients with giant cell arteritis: analysis with quantitative real time polymerase chain reaction. AB - OBJECTIVE: To evaluate the role of parvovirus B19 (B19), varicella zoster virus (VZV), and human herpes virus 6 (HHV-6) in the aetiopathology of giant cell arteritis (GCA). METHODS: Temporal artery biopsy specimens from 57 patients with GCA and 56 controls were investigated. DNA was obtained by biopsy, and quantitative real time polymerase chain reaction assay performed to establish the prevalence and viral load of B19, VZV, and HHV-6. Amplification of the human beta globin gene was used as internal positive control. RESULTS: (a) B19 was detected in 31/57 (54%) patients (median viral load 45.2 (25th-75th centiles 0-180.2) copies/microg DNA) v 21/56 (38%) controls (median viral load 0 (0-66.7) copies/microg of DNA; p = 0.07 for DNA prevalence, p = 0.007 for viral load. Among 31 B19 positive samples, 21 (68%) patients with biopsy proven GCA had >10(2) B19 copies/microg of DNA v 5/21 (24%) controls; p = 0.001. (b) No significant difference was found for VZV (p = 0.94 for DNA prevalence; p = 0.76 for viral load) and HHV-6 (p = 0.89 for DNA prevalence; p = 0.64 for viral load) in the GCA group compared with controls. CONCLUSION: B19 may have a role in the aetiopathology of GCA, particularly in those patients with high viral load; no evidence was found for VZV and HHV-6. PMID- 15834061 TI - Cerebral vasculitis as a primary manifestation of systemic lupus erythematosus. PMID- 15834062 TI - C reactive protein: protecting from lupus in familial Mediterranean fever. PMID- 15834063 TI - Iron deficiency anaemia in chronic inflammatory rheumatic diseases: low mean cell haemoglobin is a better marker than low mean cell volume. PMID- 15834064 TI - Reactivation of a latent precore mutant hepatitis B virus related chronic hepatitis during infliximab treatment for severe spondyloarthropathy. PMID- 15834065 TI - Use of herbal remedies and potential drug interactions in rheumatology outpatients. PMID- 15834066 TI - Anti-cyclic citrullinated peptide antibodies in patients with primary Sjogren's syndrome. PMID- 15834067 TI - Brucellosis as a cause of carpal tunnel syndrome. PMID- 15834068 TI - Polymorphism at position -308 of the tumour necrosis factor alpha gene and rheumatoid arthritis pharmacogenetics. PMID- 15834069 TI - Frozen shoulder. PMID- 15834070 TI - Disparities in health according to socioeconomic status. PMID- 15834071 TI - New issues in tuberculosis. PMID- 15834072 TI - Hydroxychloroquine screening. PMID- 15834073 TI - Vision restoration therapy. PMID- 15834074 TI - Minding the gap. PMID- 15834075 TI - How to assess the prevalence of trachoma. PMID- 15834076 TI - Limbal epithelial crypts: a novel anatomical structure and a putative limbal stem cell niche. AB - BACKGROUND/AIMS: There is substantial evidence that mammalian epithelial stem cells are located within well defined niches. Although the corneoscleral limbus is acknowledged as the site of corneal epithelial stem cells no anatomical niche for such cells has yet been described. The authors undertook to re-evaluate the microanatomy of the limbus in order to identify possible sites that may represent a stem cell niche. METHODS: Systematic serial 5-7 microm sections of human corneoscleral segments obtained from cadaver donors, were examined. The sections were stained with haematoxylin and eosin or toludine blue. Sections with specific areas of interest were further examined immunohistologically for the corneal epithelial marker cytokeratin 14 and the "stem cell" marker ABCG2 transporter protein. RESULTS: Distinct anatomical extensions from the peripheral aspect of the limbal palisades were identified. These consist of a solid cord of cells extending peripherally or circumferentially. The cells stained positive for CK14 and ABCG2. CONCLUSIONS: A novel anatomical structure has been identified at the human limbus, which demonstrates characteristics of being a stem cell niche. The authors have termed this structure the limbal epithelial crypt. PMID- 15834078 TI - Clinical evaluation of the pressure phosphene tonometer in patients with glaucoma. AB - AIM: To evaluate the reliability of the pressure phosphene tonometer in comparison with the Goldmann applanation tonometer. METHODS: 45 consecutive patients with glaucoma (78 eyes) participated in the study. Eyes with previous eye surgery, a documented peripheral visual field defect, a refractive error of more than 5 dioptres, and patients who were unable to understand the procedure were excluded from the study. Intraocular pressure was measured with a pressure phosphene tonometer by one examiner and with a Goldmann applanation tonometer by two other examiners no more than 15 minutes apart. A second series of measurements was performed several weeks later on 34 patients (59 eyes). There was no communication between examiners or between examiner and patient regarding test results. RESULTS: No statistically significant correlation was found between the applanation tonometry values and those obtained with a pressure phosphene tonometer. CONCLUSION: The pressure phosphene tonometer is not suitable for reliably measuring intraocular pressure. PMID- 15834077 TI - Anti-TNF-alpha therapy for sight threatening uveitis. AB - AIM: To describe the effect of additional treatment with anti-TNF-alpha therapy in a case series of 13 patients with serious sight threatening uveitis. METHODS: 13 patients with serious sight threatening uveitis were included, of whom six had Behcet's disease, five had idiopathic posterior uveitis, one had sarcoidosis, and one birdshot retinochoroiditis. Onset and course of ocular inflammation, inflammatory signs, and visual acuity were assessed. Patients were treated with 200 mg (approximately 3 mg/kg) infliximab infusion. Repeat infusions were given based on clinical response. RESULTS: Infliximab treatment resulted in an effective suppression of ocular inflammation in all patients. In patients with non-Behcet's disease uveitis visual acuity in six out of eight improved or was stable. In patients with Behcet's disease visual acuity in five out of six improved or was stable. CONCLUSION: Anti-TNF-alpha treatment may be of value in the treatment of uveitis, and in patients with Behcet's disease, leading to suppression of ocular inflammation, vasculitis, and improvement of vision in the majority. Based on these results a controlled masked study is warranted. PMID- 15834079 TI - Intraocular pressure variability in patients who reached target intraocular pressure. AB - AIM: To assess the intraocular pressure (IOP) variability in patients with primary open angle glaucoma (POAG) under clinical treatment who reached an established target pressure based on isolated office readings. METHODS: Retrospective analysis of 65 eyes from 65 POAG patients under clinical therapy who submitted to modified diurnal tension curve (mDTC) (measurements at every 3 hours between 8 am and 5 pm) followed by a water drinking test (WDT). All subjects had established target IOP < or =15 mm Hg at 11 am or 2 pm. IOP variability during mDTC or WDT was evaluated. RESULTS: mDTC revealed IOP measurements > or =17 mm Hg in 16 of 65 eyes (24.6%). Nine eyes (13.8%) presented values > or =18 mm Hg. The highest IOP detected by mDTC was 20 mm Hg in one patient (1.5%). WDT demonstrated IOP values > or =17 mm Hg in 32 of 65 eyes (49.2%). 22 eyes (33.8%) presented values > or =18 mm Hg after water ingestion. Moreover, IOP levels > or =20 mm Hg were observed in 14 eyes (21.5%). CONCLUSION: A great percentage of POAG patients undergoing clinical treatment and with IOP control based on single office measurement present significantly higher IOP measurements when performing mDTC and, especially, the WDT. PMID- 15834080 TI - A novel index for predicting intraocular pressure reduction following cataract surgery. AB - AIM: The results of a study designed to investigate the predictive value of preoperative anterior chamber depth (ACD) and intraocular pressure (IOP) are reported. The relation between these factors and their effect on the reduction in IOP following phacoemulsification cataract surgery was also studied. METHODS: The ACD and IOP were prospectively measured in 103 non-glaucomatous eyes of 103 patients who underwent uneventful phacoemulsification and posterior chamber intraocular lens (PCIOL) implantation. Other data which were recorded included best corrected visual acuity, axial length, lens thickness, and severity of lens opacity. RESULTS: The ACD increased by a mean (SD) of 1.10 (0.44) mm (p<0.00001) and this increase was significantly and inversely related to preoperative ACD (r(2) = 68%; p<0.01). IOP dropped by a mean of 2.55 (1.78) mm Hg following cataract surgery (p<0.0001), and this reduction was significantly and positively related to preoperative IOP (r(2) = 56%; p<0.01), and significantly and inversely related to preoperative ACD (r(2) = 21%; p<0.01). A novel ratio, the pressure to depth (PD) ratio (preoperative IOP/preoperative ACD), was found to be significantly and positively related to the surgically induced reduction in IOP (r(2) = 73%; p<0.01), and IOP was reduced by > or =4 mm Hg in all patients with a PD ratio >7. CONCLUSION: The reduction in IOP following cataract surgery was found to be positively related to preoperative IOP, and inversely related to preoperative ACD. Furthermore, these results indicate that a novel index, the PD ratio, is strongly predictive for IOP reduction following cataract extraction, and may prove useful in surgical decision making. PMID- 15834081 TI - Effects of the combination of bimatoprost and latanoprost on intraocular pressure in primary open angle glaucoma: a randomised clinical trial. AB - AIMS: To evaluate the effect of the combination of bimatoprost and latanoprost on intraocular pressure (IOP) in primary open angle glaucoma (POAG). METHODS: An open label randomised clinical trial was conducted, which included 18 glaucomatous patients (36 eyes). In the first 4 weeks, latanoprost 0.005% was prescribed for both eyes of the patients and any other antiglaucoma medication was discontinued. In the next 4 weeks (phase 1), bimatoprost 0.03% was combined with latanoprost in one randomly assigned eye (case eye) of each patient. In the next 4 weeks (phase 2), bimatoprost was discontinued in the case eyes, while bimatoprost was substituted for latanoprost in the fellow eye (control eye). The IOP was measured at the end of the first 4 weeks (baseline measurement) and weekly during phases 1 and 2. RESULTS: In the case eyes, the mean IOP increased along the first phase (1.8 mm Hg; p = 0.006) when compared to baseline measurements. The IOP returned to previous values after discontinuation of bimatoprost in phase 2. In the control eyes, the mean IOP did not change throughout the study. CONCLUSION: The combination of bimatoprost and latanoprost in POAG increases the IOP and should not be considered as a therapeutic option. PMID- 15834082 TI - 28,000 Cases of age related macular degeneration causing visual loss in people aged 75 years and above in the United Kingdom may be attributable to smoking. AB - BACKGROUND: Age related macular degeneration (AMD) causing visual impairment is common in older people. Previous studies have identified smoking as a risk factor for AMD. However, there is limited information for the older population in Britain. METHODS: Population based cross sectional analytical study based in 49 practices selected to be representative of the population of Britain. Cases were people aged 75 years and above who were visually impaired (binocular acuity <6/18) as a result of AMD. Controls were people with normal vision (6/6 or better). Smoking history was ascertained using an interviewer administered questionnaire. RESULTS: After controlling for potentially confounding factors, current smokers were twice as likely to have AMD compared to non-smokers (odds ratio 2.15, 95% CI 1.42 to 3.26). Ex-smokers were at intermediate risk (odds ratio 1.13, 0.86 to 1.47). People who stopped smoking more than 20 years previously were not at increased risk of AMD causing visual loss. Approximately 28,000 cases of AMD in older people in the United Kingdom may be attributable to smoking. CONCLUSION: This is the largest study of the association of smoking and AMD in the British population. Smoking is associated with a twofold increased risk of developing AMD. An increased risk of AMD, which is the most commonly occurring cause of blindness in the United Kingdom, is yet another reason for people to stop smoking and governments to develop public health campaigns against this hazard. PMID- 15834083 TI - An interinstitutional comparative study and validation of computer aided drusen quantification. AB - AIMS: To assess the portability and clinical applicability of a software program based on Photoshop (Adobe Systems Inc, San Jose, CA, USA) for digital drusen quantification. METHODS: Independent graders from the Digital Fundus Photo Reading Center of Columbia University and King's College Hospital used macular background levelling software to quantify the percentage of drusen in the central and middle Wisconsin subfields. 100 images of consecutive patients with choroidal neovascularisation in one eye and significant drusen in the other eye were analysed to determine suitability, and 10 were chosen for assessment by this software. RESULTS: Of the 10 images used in the interinstitutional validation, the random effects ANOVA for the central and middle subfields showed a high degree of interobserver agreement. The ICC for interobserver reliability was 0.83 (95% CI: 67 to 95) for the central subfield and 0.84 (95% CI: 69 to 99) for the middle subfield. Overall agreement with the manual grading results was good and the within patient coefficient of variation was about 20% for all the pairwise comparisons between observers and the manual stereo gradings. Of the 100 images used to assess practical applicability of the software, 79 were suitable for semiautomated analysis. 13 had extensive mixed retinal pigment epithelial (RPE) changes limiting drusen identification, five had a significant number of reticular drusen, which are poorly identified by the software, and three had multiple small areas of RPE atrophy, which are difficult to distinguish from drusen. CONCLUSIONS: The software was successfully used by two institutions demonstrating portability, with good correlation between graders and to the manual stereo grading. Digital drusen quantification was possible in 79% of the images analysed. PMID- 15834084 TI - Radial optic neurotomy for ischaemic central vein occlusion. AB - BACKGROUND/AIMS: Ischaemic central retinal vein occlusion (CRVO) accounts for 20 50% of all CRVO. No treatment has been proved to be effective. The efficacy of radial optic neurotomy (RON) was evaluated in eyes with ischaemic CRVO. METHODS: 10 patients with ischaemic CRVO underwent RON. After pars plana vitrectomy, a microvitreoretinal blade was used to incise the scleral ring, cribriform plate, and adjacent sclera at the nasal edge of the optic disc. Best corrected visual acuity (BCVA), intraocular pressure (IOP), fluorescein angiography (FA), multifocal electroretinography (mfERG), and optical coherence tomography (OCT) were measured preoperatively and at 1, 3, and 6 months postoperatively. RESULTS: No visual improvement was noted in the eyes that underwent RON. FA and mfERG showed no increase in retinal perfusion or retinal function postoperatively. Mean macular central thickness changed from 841 (SD 170) mum preoperatively to 162 (SD 34) microm at the sixth postoperative month. One patient had retinal central artery perforation intraoperatively. One patient developed neovascular glaucoma. CONCLUSION: RON in ischaemic CRVO did not improve visual function (by mfERG) or visual acuity although macular thickness did improve. This technique may be associated with potential risks. Randomised studies are needed to corroborate these results. PMID- 15834085 TI - Optical coherence tomography characterisation of idiopathic central serous chorioretinopathy. AB - AIM: To describe retinal findings in patients with idiopathic central serous chorioretinopathy (ICSC) as assessed by optical coherence tomography (OCT), and to compare them to fluorescein angiography (FA) findings. METHODS: Case series of 39 eyes from 36 patients with ICSC. Complete ophthalmological examination, last generation OCT (StratusOCT, Software version 3.1) and FA were performed. Six radial scans using OCT were performed and repeated. Singular findings were recorded, OCT images were measured and the results compared with those of FA. The main outcome measures were FA and OCT findings. RESULTS: Two patterns of distinct OCT findings are described. In the first one, an optically empty vaulted area of different heights was observed under the neurosensory retina in 36 eyes, being related to fluorescein filled areas; in 35 of them, highly characteristic small bulges could be observed protruding from the retinal pigment epithelium (RPE), angiographically related to leaking spots. In the second pattern, three eyes showed an almost semicircular space under the RPE, with thinner overlying retina. CONCLUSIONS: OCT may offer a new approach to the staging and knowledge of ICSC, and may help the understanding of the mechanisms of the disease. PMID- 15834086 TI - Evaluation of internet websites about retinopathy of prematurity patient education. AB - BACKGROUND/AIMS: The success of the treatment in patients with retinopathy of prematurity (ROP) is mainly associated with timely diagnosis and appropriate management. Information dissemination is crucial for the outcome of ROP. This study aimed to evaluate the quality of the information about ROP available for patients on the internet. METHODS: Cross sectional study. In March 2004 the ROP information available on the internet was evaluated using two search engines (MetaCrawler and MSN) and four key terms ("retinopathy of prematurity," "premature eye," "premature retina," and "ROP"). The quality of each website was evaluated using a score system. The sites were classified as academic, organisational, or commercial. Readability, general quality of the website (based on ownership, purpose, authorship, author qualification, attribution, interactivity, and currency), and quality of the content specific to ROP (definition, causes, epidemiology, risk factors, diagnosis, classification, treatment, and prognosis) were analysed. RESULTS: Of 114 websites evaluated, 40 were included. 10 sites (25.0%) were academic, eight (20.0%) organisational, and 22 (55.0%) commercial. In the majority of the sites (62.5%) the ROP information was fair or poor. CONCLUSIONS: A large amount of information about ROP is available on the internet. However, most websites were considered incomplete. PMID- 15834087 TI - Threshold Amsler grid as a screening tool for asymptomatic patients on hydroxychloroquine therapy. AB - BACKGROUND/AIMS: Patients taking hydroxychloroquine (HCQ) are at risk of developing classic bull's eye maculopathy. Currently, the standard Amsler grid (AG) is one of the most useful methods to identify such lesions. However, AG is a suprathreshold target and may not detect relative central scotomas. The aim of this study was to determine if the threshold Amsler grid (TAG) test, which varies light transmission through two cross polarising filters, allows increased detection of scotomas caused by HCQ toxicity. METHODS: 56 rheumatological patients taking HCQ and 12 similar patients not taking HCQ were tested by AG, red Amsler grid (RAG), and TAG. RESULTS: No scotomas were observed in patients never treated with HCQ. Among patients who had been treated with HCQ, AG revealed scotomas in two of 56 (3.64%) patients; in contrast, six (10.7%) and 37 (66.1%) scotomas were identified by RAG and TAG testing respectively. Additionally, the average area of each scotoma detected by all three methods expanded from 34.5 square degrees of central field loss on AG testing to 71 square degrees on RAG and 117 on TAG. CONCLUSION: By decreasing the perceived luminance of the suprathreshold AG, TAG testing provides a novel alternative to detect shallow scotomas and areas of depressed retinal activity secondary to HCQ toxicity. PMID- 15834088 TI - Long term outcome of trichiasis surgery in the Gambia. AB - BACKGROUND: Trichiasis surgery is believed to reduce the risk of losing vision from trachoma. There are limited data on the long term outcome of surgery and its effect on vision and corneal opacification. Similarly, the determinants of failure are not well understood. METHODS: A cohort of people in the Gambia who had undergone surgery for trachomatous trichiasis 3-4 years earlier was re assessed. They were examined clinically and the conjunctiva was sampled for Chlamydia trachomatis polymerase chain reaction (PCR) and general bacterial culture. RESULTS: In total, 141/162 people were re-examined. Recurrent trichiasis was found in 89/214 (41.6%) operated eyes and 52 (24.3%) eyes had five or more lashes touching the globe. Corneal opacification improved in 36 of 78 previously affected eyes. There was a general deterioration in visual acuity between surgery and follow up, which was greater if new corneal opacification developed or trichiasis returned. Recurrent trichiasis was associated with severe conjunctival inflammation and bacterial infection. C trachomatis was detected in only one individual. CONCLUSIONS: Recurrent trichiasis following surgery is a common potentially sight threatening problem. Some improvement in the cornea can occur following surgery and the rate of visual loss tended to be less in those without recurrent trichiasis. The role of conjunctival inflammation and bacterial infection needs to be investigated further. Follow up of patients is advised to identify individuals needing additional surgical treatment. PMID- 15834089 TI - Corneal sensation after myopic and hyperopic LASIK: clinical and confocal microscopic study. AB - AIM: To assess the long term (1 year) effect of myopic and hyperopic LASIK on corneal sensation and innervation. METHODS: 83 eyes of 43 patients having LASIK were evaluated. According to the preoperative spherical equivalent, the eyes were divided into three groups: group 1, myopia from -0.75 to -6.00 D; group 2, myopia from -6.25 to -11.50 D; and group 3, hyperopia from 1.25 to 5.00 D. Corneal sensation was measured and in vivo confocal microscopy (IVCM) was done at the central cornea before, and at 1 month, 3 months, 6 months, and 1 year after LASIK. RESULTS: The mean corneal sensation in group 1 was greater than in groups 2 and 3 at all postoperative measurements. The difference between group 1 on one hand and groups 2 and 3 on the other hand was statistically significant at 1 month and 3 months after LASIK and was not statistically significant afterwards. IVCM study of 27 eyes revealed that the number and length of nerve fibre bundles in the sub-basal region decreased after LASIK and was significantly lower at all times after surgery despite the return of corneal sensation to preoperative level. CONCLUSION: After LASIK, central corneal sensitivity is decreased for as long as 6 months or more. The results suggest that lamellar cutting of the cornea during LASIK impairs corneal sensitivity and is related to the ablation depth. The diameter of ablation too may contribute to this drop in sensitivity. The return of corneal sensations does not directly correlate with the regeneration of nerve fibres as determined by confocal imaging. Sensations return to normal values before complete restoration of normal innervation if this indeed ever occurs. PMID- 15834090 TI - Efficiency of blood culture bottles for the fungal sterility testing of corneal organ culture media. AB - BACKGROUND/AIM: The consequences of fungal contamination of an organ cultured cornea, though exceptional, are often disastrous for the recipient. Consequently, eye banks often quarantine corneas for 10 days or more before passing them for grafting. This period, though detrimental to the endothelial cell density of the delivered cornea, is necessary to detect contamination using conventional microbiological methods. The authors previously validated the use of a pair of aerobic and anaerobic blood bottles for sensitive and rapid detection of bacteria. To allow a short quarantine period, it remained only to optimise detection of fungi. The authors aimed to compare sensitivity and rapidity of fungal contamination detection by three methods: blood bottles, Sabouraud, and daily visual inspection of the organ culture medium. METHODS: Four inocula (10(6), 10(4), 10(2), 10 colony forming unit (CFU) per ml) of 11 fungi (Candida albicans, C tropicalis, C glabrata, Saccharomyces cerevisiae, Rhodotorula rubra, Cryptococcus neoformans, Fusarium oxysporum, Aspergillus niger, A fumigatus, A flavus, Acremonium falciforme) were inoculated in a commercial organ culture medium containing a coloured pH indicator (CorneaMax, Eurobio, Les Ulis, France). The real live fungal inoculum was verified immediately after inoculation. After 48 hours at 31 degrees C, samples of the contaminated media were inoculated in three blood bottles: Bactec Aerobic/F, Bactec Mycosis IC/F, and Bactec Myco/F Lytic (Becton Dickinson, Le Pont de Claix, France), then placed in a Bactec 9240 rocking automat, and in four Sabouraud media (solid and liquid, 28 degrees C and 37 degrees C) with daily observation. Contaminated organ culture media were also checked daily for any change in turbidity and/or colour. Experiments were performed in triplicate. RESULTS: Mycosis IC/F and Myco/F Lytic bottles were neither faster nor more sensitive than the aerobic bottle. The three methods were positive for all inocula, even the lowest (viable inoculum below 10 CFU/ml for each fungus). Contamination was detected within 24 hours by the aerobic bottles in 91% (40/44), by Sabouraud in 98% (43/44) (no significant difference) and by visual inspection in 66% of cases (29/44) (p<0.001 with the two others). Maximum times to detection were 46, 48 and 72 hours respectively. CONCLUSION: This study further counters the preconception that fungal contamination is hard to detect in corneal organ culture media. This study is the last step in validating the use of a pair of blood bottles for the sterility testing of organ culture media, this time for fungi. Their use should make it possible to shorten microbiological quarantine and thus deliver corneas with higher endothelial cell density, without increasing the risk of recipient contamination. PMID- 15834091 TI - Infective keratitis in older patients: a 4 year review, 1998-2002. AB - BACKGROUND/AIM: There are few clinical series in the literature of infective keratitis in the elderly, even though this age group constitutes a significant proportion of those affected by this condition. The authors aimed to determine the incidence and risk factors for infective keratitis in those over 60 years, the causative organisms, antibiotic susceptibilities, visual and tectonic outcome, and surgical intervention rate. METHODS: A retrospective review of all patients aged 60 years and over admitted to the Sydney Eye Hospital with a diagnosis of infective keratitis, between September 1998 and December 2002. RESULTS: 190 patients were identified with a mean age of 75.5 (SD 9.6) years (range 60-101). Local risk factors were found in 93.7%, and systemic risk factors in 27.9%. Organisms were cultured in 62.8%, and 7.9% had positive herpes simplex virus (HSV) polymerase chain reaction (PCR). Perforation or severe thinning occurred in 36% overall, but in 80% with positive HSV PCR. Acute surgical intervention was required in 43.7%, with acute penetrating keratoplasty performed in 17.9%, and 8.9% required evisceration. Mean presenting visual acuity was 1.82 (SD 1.24), equivalent to 6/300, excluding 26.3% with vision of light perception (LP) or worse. Mean final visual acuity was 1.24 (SD 1.16), equivalent to 6/100, excluding 19.5% with vision of LP or worse (p<0.0005). CONCLUSIONS: The elderly represent a distinct clinical group in the context of microbial keratitis. Predisposing factors are very common, they present with poor vision, have a high complication and surgical intervention rate, and a poor visual outcome compared to younger patients. The microbiological spectrum is similar to younger age groups, except that HSV is more common and may increase the risk of severe corneal thinning and perforation. Most bacterial isolates remain sensitive to currently available antibiotic preparations. PMID- 15834092 TI - Filtering bleb function after clear cornea phacoemulsification: a prospective study. AB - AIM: To evaluate the influence of clear cornea phacoemulsification on filtering bleb morphology, function, and intraocular pressure (IOP) in glaucomatous eyes with previously successful filtering surgery. METHODS: The clinical course of 30 patients (30 eyes) who underwent clear cornea phacoemulsification after successful filtering glaucoma surgery was prospectively evaluated. Mean IOP and filtering bleb morphology (standardised assessment criteria and score 0-12, 12 = optimum) were determined before surgery, and 3 days, 6 months, and 12 months after surgery. The control group consisted of 36 patients with glaucoma after clear cornea phacoemulsification without previous filtering surgery. RESULTS: Mean IOP increased after phacoemulsification by about 2 mm Hg (preoperatively 14.28 (SD 3.71) mm Hg, 12 months postoperatively 16.33 (3.31) mm Hg, p = 0.006). 15 patients (50%) showed an IOP increase of >2 mm Hg, 11 patients (36.7%) had no IOP difference (within 2 mm Hg), and in four patients (13.3%) IOP decreased >2 mm Hg. Mean score of filtering bleb morphology 1 year after surgery decreased from 9.5 to 9.0 (p = 0.154). In three of 30 preoperatively IOP regulated eyes the postoperative IOP was 21 mm Hg. The control group showed an average IOP decrease of 2.01 mm Hg (p = 0.014) 12 months after cataract surgery. CONCLUSION: An increase in IOP was found 1 year after phacoemulsification in half of the filtered glaucomatous eyes. IOP in glaucomatous eyes without previous filtering surgery decreased in the same period. Cataract extraction using clear cornea phacoemulsification may be associated with a partial loss of the previously functioning filter and with an impairment of filtering bleb morphology. PMID- 15834093 TI - The origins of polypoidal choroidal vasculopathy. AB - BACKGROUND/AIM: There are two theories on the pathogenesis of polypoidal choroidal vasculopathy (PCV): variants in choroidal neovascularisation (CNV) and inner choroidal vessel abnormalities. On indocyanine green angiography (IGA) with a video camera system, PCV has a characteristic appearance, but inadequate image quality has made detailed interpretation difficult. This study aims to improve imaging, using confocal scanning laser ophthalmoscopy (SLO), to elucidate the pathogenesis of PCV. METHODS: High speed IGA with confocal SLO of 45 eyes (44 patients) showed typical PCV findings of a branching vascular network and polypoidal lesions. RESULTS: Vessels comprising branching networks began to fill simultaneously with the surrounding choroidal arteries in 38 eyes. Small numbers of vessels filling within a branching network, in the arterial and arteriovenous phases of IGA, showed focal dilatation, constriction, and tortuousity. Vessel abnormalities, corresponding to polypoidal lesions, existed within a network in eight eyes and included loops similar in calibre to network vessels, and numerous microaneurysmal dilatations of small vessels. Vessel pulsation was seen in 24 eyes. CONCLUSION: PCV is caused by inner choroidal vessel abnormalities, not CNV. PMID- 15834094 TI - ANCA associated pauci-immune retinal vasculitis. AB - BACKGROUND: Antinuclear cytoplasmic antibodies (ANCA) are useful diagnostic serological markers for the most common forms of necrotising vasculitis. ANCA associated vasculitides represent distinctive clinicopathological categories--for example, Wegener's granulomatosis, Churg-Strauss syndrome, microscopic polyangiitis, and idiopathic necrotising crescentic glomerulonephritis, collectively known as the small vessel pauci-immune vasculitides. METHOD: Three cases of ANCA associated pauci-immune retinal vasculitis are described. Their systemic features are described and the clinical significance of ANCA as a diagnostic test in relation to retinal vasculitis discussed. RESULTS: These three cases represent a spectrum of clinical features associated with retinal vasculitis. Two cases have evolved into clinical recognisable entities as microscopic polyangiitis. Adherence to the international consensus statement on testing and reporting of ANCA is recommended and the authors speculate that the incidence of microscopic polyangiitis may be underestimated because of the under recognition of systemic involvement in patients with retinal vasculitis. CONCLUSION: The receipt of a positive ANCA result should always raise the suspicion of a pauci-immune systemic vasculitis and prompt appropriate investigation. The authors emphasise the importance of the evaluation of systemic features in these patients with retinal vasculitis, enabling earlier recognition and thereby preventing significant morbidity and mortality. PMID- 15834095 TI - Causes of severe visual impairment and blindness in schools for visually handicapped children in Iran. AB - AIMS: This survey was conducted on children in schools for the blind in Tehran (from 2002 to 2003) to determine the causes of severe visual impairment and blindness and to identify preventable and treatable conditions. METHODS: The study was performed on 362 students at different grades in three schools for the blind. Patient sex, age, family history of blindness or low vision, visual acuity, causes of blindness, and treatable and preventable conditions were studied. RESULTS: Of the 362 cases, 210 (58%) were boys and 152 (42%) were girls. Mean age was 13.5 (SD 4) years. Severe visual loss was seen in 80.9%. Retinal diseases were the most common cause for low vision (51%); cataract, optic nerve atrophy, corneal and anterior segment diseases, glaucoma, anophthalmia, and globe malformations were other major causes of blindness. Treatable aetiologies and positive family history of blindness were seen in 25.7% and 36% of the patients, respectively. The incidence of preventable diseases, excluding familial disorders, was low. CONCLUSION: In addition to the prevention and treatment of some conditions, premarital genetic counselling and family planning control in families with inherited diseases could decrease the number of blind children in the future in Iran. PMID- 15834096 TI - How patients experience progressive loss of visual function: a model of adjustment using qualitative methods. AB - BACKGROUND: People with retinitis pigmentosa (RP) experience functional and psychological challenges as they adjust to progressive loss of visual function. The authors aimed to understand better the process of adjusting to RP in light of the emotional suffering associated with this process. METHODS: Adults with RP were recruited from the Foundation Fighting Blindness and the Wilmer Eye Institute in Baltimore. Focus groups and semistructured interviews addressed the process of adjusting to RP and were audiotaped and transcribed. The transcripts were analysed qualitatively in order to generate a model of adjustment. RESULTS: A total of 43 individuals participated. It was found that, on diagnosis, people with RP seek to understand its meaning in their lives. Mastering the progressive functional implications associated with RP is contingent upon shifting personal identity from a sighted to a visually impaired person. In this sample, six participants self identified as sighted, 10 self identified as in transition, and 27 self identified as visually impaired. This adjustment process can be understood in terms of a five stage model of behaviour change. CONCLUSIONS: The proposed model presents one way to understand the process of adjusting to RP and could assist ophthalmologists in meeting their moral obligation to lessen patients' suffering, which arises in the course of their adjustment to progressive loss of visual function. PMID- 15834097 TI - The development of the Indian vision function questionnaire: field testing and psychometric evaluation. AB - OBJECTIVE: To develop and evaluate the acceptability, reliability, validity, and responsiveness of the Indian vision function questionnaire (IND-VFQ). METHODS: Problem statements from previous qualitative studies were reduced to a 45 item interviewer administered questionnaire representing three a priori domains (general functioning, psychosocial impact, and visual symptoms) which was evaluated in patients with cataract (n = 420), glaucoma (n = 120), diabetic retinopathy, or age related macular degeneration (n = 120) and normal controls (n = 120). Standard methods were used for item reduction and to evaluate psychometric properties. RESULTS: Psychometric item reduction produced a 33 item questionnaire. Psychometric evaluation showed that two of the three scales (psychosocial impact and visual symptoms) had good acceptability, and that all three scales showed high internal consistency (alpha >0.80; item-total correlations 0.54-0.86) and test-retest reliability (>0.89). All three scales showed moderate evidence of convergent and discriminant validity. Responsiveness, assessed in cataract patients (n = 120) before and after surgery, was good for all three scales (effect sizes >1). CONCLUSIONS: The IND-VFQ33 is a psychometrically sound measure of vision function addressing a gap in patient defined measures of vision function developed in populations living in low income countries. PMID- 15834098 TI - Penetration of moxifloxacin into the human aqueous humour after oral administration. AB - AIMS: To determine the pharmacokinetics of moxifloxacin, a new generation fluoroquinolone, in the anterior chamber of the human uninflamed eye. METHODS: 35 patients undergoing cataract surgery received two doses of 400 mg of oral moxifloxacin with a 12 hour interval and were divided into six groups. Moxifloxacin levels in aqueous humour and serum were determined by a microbiological agar well diffusion technique at 2, 4, 6, 8, 10, and 12 hours after the second dose in each group respectively. RESULTS: Mean moxifloxacin levels in the anterior chamber were 1.20 (SD 0.35) microg/ml at the 2 hours group, 1.22 (0.48) microg/ml at the 4 hours group, 1.20 (0.45) microg/ml at the 6 hours group, 1.58 (0.38) microg/ml at the 8 hours group, 1.37 (0.44) microg/ml at the 10 hours group, and 1.23 (0.55) microg/ml at the 12 hours group. The mean ratio of aqueous to serum moxifloxacin level was 38%. CONCLUSION: Moxifloxacin penetrates well into the anterior chamber of the human uninflamed eye after oral administration, reaching early significant levels, which are maintained for at least 12 hours and are much higher than the MIC(90) values of Gram positive and Gram negative pathogens commonly implicated in intraocular infections with the exceptions of fluoroquinolone resistant staphylococci, MRSA, and Pseudomonas aeruginosa. PMID- 15834100 TI - Disappearance of eyelid xanthelasma following oral simvastatin (Zocor). PMID- 15834099 TI - Corneal graft rejection occurs despite Fas ligand expression and apoptosis of infiltrating cells. AB - BACKGROUND/AIMS: Constitutive expression of Fas ligand (CD95L) protects the eye against cell mediated immune responses by inducing apoptosis in infiltrating Fas bearing T cells. This study was designed to examine Fas ligand expression on acutely rejecting rat corneal grafts and to investigate the kinetics of induction of apoptosis in infiltrating leucocytes. METHODS: Orthotopic penetrating corneal transplantation was performed between genetically disparate inbred rats. Fas ligand expression and the phenotype of infiltrating leucocytes were examined by immunohistochemistry. Apoptotic nuclei were visualised in sections of normal rat cornea, rejecting allografts, and time matched isografts by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labelling (TUNEL) and quantified by video image analysis. Staining with Hoechst dye 33258 was used to confirm the presence of apoptotic nuclei. RESULTS: Fas ligand was expressed on corneal endothelial and epithelial cells during acute corneal graft rejection. At all time points examined, including as early as the fifth postoperative day, the cells infiltrating both corneal isografts and allografts were TUNEL positive. By the 15th postoperative day, over 90% of all nuclei, many of which were T cells, were apoptotic. CONCLUSION: Expression of Fas ligand is not downregulated on the cornea during allograft rejection and infiltrating leucocytes in both isografts and allografts die rapidly in situ. Despite the death of the cells believed to be responsible for rejection, isografts survive indefinitely whereas allografts are irreparably damaged. PMID- 15834101 TI - Inverse globe retraction syndrome complicating recurrent pterygium. PMID- 15834102 TI - New onset diplopia: 14 years after retinal detachment surgery with a hydrogel scleral buckle. PMID- 15834103 TI - Seeing is not believing. PMID- 15834104 TI - Radial optic neurotomy in combined cilioretinal artery and central retinal vein occlusion. PMID- 15834105 TI - Value based medicine. PMID- 15834106 TI - Cystoid macular oedema with trypan blue use. PMID- 15834107 TI - The tau mutation in the Syrian hamster differentially reprograms the circadian clock in the SCN and peripheral tissues. AB - The hypothalamic suprachiasmatic nuclei (SCN), the principal circadian oscillator in mammals, are synchronized to the solar day by the light-dark cycle, and in turn, they coordinate circadian oscillations in peripheral tissues. The tau mutation in the Syrian hamster is caused by a point mutation leading to a deficiency in the ability of Casein Kinase 1epsilon to phosphorylate its targets, including circadian PER proteins. How this accelerates circadian period in neural tissues is not known, nor is its impact on peripheral circadian oscillators established. We show that this mutation has no effect on per mRNA expression nor the nuclear accumulation of PER proteins in the SCN. It does, however, accelerate the clearance of PER proteins from the nucleus to an extent sufficient to explain the shortened circadian period of behavioral rhythms. The mutation also has novel, unanticipated consequences for circadian timing in the periphery, including tissue-specific phase advances and/or reduced amplitude of circadian gene expression. The results suggest that the tau mutation accelerates a specific phase, during mid-late subjective night of the SCN circadian feedback loop, rather than cause a global compression of the entire cycle. This reprogrammed output from the clock is associated with peripheral desynchrony, which in turn could account for impaired growth and metabolic efficiency of the mutant. PMID- 15834108 TI - The thymus is similar to the testis in its pattern of circadian clock gene expression. AB - The molecular basis for the circadian clock in mammals consists of a number of genes and proteins that form transcription-translation feedback loops. These loops result in a 24-h rhythm in the expression of mRNA and protein levels. Although the anatomical site of the central circadian clock is the SCN of the hypothalamus, all of the circadian clock genes are expressed in tissues other than the brain. Moreover, cyclic gene and protein expression occurs in most of these tissues. The best known exception to this rule is the testis, which shows constant rather than cyclic expression of circadian clock genes. Indeed, the testis of multiple animal species displays constant circadian clock gene expression. In recent work, the authors showed that the thymus is similar to the testis in that expression of circadian clock genes is either constant over a 24-h period or cycles with a dampened amplitude, depending on which gene is examined. In the current study, they extend and confirm their findings regarding noncyclic circadian clock gene and protein expression in the testis and the thymus. More important, they also show that expression of these genes in both testis and thymus does not depend on the transcriptional activator, CLOCK, which is necessary for cyclic gene expression in the SCN and in other tissues. These results extend the molecular similarities between the thymus and the testis and suggest that similar mechanisms are at work for regulating expression of circadian clock genes in both tissues. One commonality between these 2 organs is that they are composed primarily of differentiating cells. The authors hypothesize that the circadian clock is not operational in immature, differentiating cells. Possibly, the clock starts in mature cells upon receipt of an initiating signal. PMID- 15834109 TI - Circadian rhythm phenotype of 5-HT7 receptor knockout mice: 5-HT and 8-OH-DPAT induced phase advances of SCN neuronal firing. AB - In vitro neuronal recordings in the SCN have clearly documented shifts in the peak of unit activity following the application of serotonergic agents, and yet selectivity issues with these very tools have limited progress in establishing the precise receptor mechanisms. As an alternative strategy, mice were bred (C57BL/6J) lacking 1 serotonin receptor, the 5-HT(7), to serve as a null background for this subtype; earlier work had documented the involvement of 5 HT(7) receptors in the phase advances elicited by 8-OH-DPAT, a mixed 5-HT(1A/7) agonist, in SCN slices prepared from rat donors. Single-unit recordings in sequential electrode passes revealed peaks of activity that occurred at nearly the same time in the knockout (KO; ZT4.2 +/- 0.6) and wild-type animals (WT; ZT4.3 +/- 0.1), where ZT0 marks the beginning of the light phase in a 12:12 LD cycle. Bath application of 8-OH-DPAT produced a phase advance in neuronal firing (2.1 +/- 0.5 h) when applied 1 circadian cycle earlier at ZT6 (10 microM, 10 min), but surprisingly, the mean phase advance in slices prepared from KO mice (2.3 +/- 0.1 h) was no different. Coapplication of 8-OH-DPAT with WAY-100,635 (10 microM), a highly selective 5-HT(1A) antagonist, significantly reduced the phase advance, both in experiments with WT and KO mice, suggesting the greater importance of this serotonin sub-type independent of genetic modification. 5-HT itself (0.5 +/-M, 10 min) at ZT6 also yielded phase advances that were indistinguishable in slices prepared from WT and KO mice (1.8 +/- 0.4 h and 2.1 +/- 0.2 h, respectively) and that were also sensitive to WAY-100,635. Unlike the pattern with 8-OH-DPAT, however, 5-HT-induced phase advances, in both WT and KO mice, were blocked by ritanserin, in this paradigm useful as a 5-HT(5A/7) antagonist (in addition to its more typical role as a 5-HT2A/2C antagonist). Serotonin antagonists when administered alone were without effect in slices from WT mice but produced significant phase shifts when administered to those from KO animals. Taken together, these results highlight the importance of the species used in establishing receptor mechanism. More provocatively, they support the involvement of multiple serotonin receptors in shifting the phase of circadian rhythms at ZT6. PMID- 15834110 TI - Season- and latitude-dependent effects of simulated twilights on circadian entrainment. AB - Groups of Syrian hamsters were exposed to LD cycles with twilight transitions and photoperiods simulating natural lighting conditions at the summer solstice (SS), equinox, and winter solstice (WS) at 41 degrees N and at the winter solstice at the Arctic Circle (WS 66 degrees N) but with daytime illuminance truncated at 10 lux (LD-twilight). Separate groups were kept under matching rectangular cycles (LD-rectangular). The inclusion of twilights affected several circadian parameters in a season-and latitude-dependent manner. The most striking difference was in the timing of activity onsets, which followed dusk in the presence of twilights but were more closely related to dawn (lights-on) in their absence. Activity offsets and midpoints were also earlier in LD-twilight than in LD-rectangular, with the differences being most pronounced under WS 66 degrees N. In LD-twilight, longer nights resulted in earlier offsets and midpoints, but in LD-rectangular, midpoints were later under long than under short nights while offsets did not vary significantly. In LD-twilight, activity duration (alpha) increased monotonically with increasing nighttime duration, but in LD rectangular, alpha was shorter under WS 66 degrees N than under WS conditions. These effects of season and latitude observed in LD-twilight were similar to those reported in animals exposed to natural illumination, while those observed in LD-rectangular differed in several respects. The presence of twilights also resulted in lower day-to-day variability in activity onset times (greater precision), supporting the earlier conclusion that twilights increase the strength of the LD zeitgeber. Free-running periods in constant darkness (DD) were shorter in LD-twilight than in LD-rectangular, especially under WS 66 degrees N, raising the possibility that the effects of twilights on the timing of the entrained activity rhythm reflect their effects on the period of that rhythm. Increasing daytime illuminance to 100 lux (WS conditions only) resulted in earlier activity offsets and midpoints and a shorter alpha but had no effect on activity onsets or on subsequent period in DD. These results indicate that exposure to low twilight illuminances alone can account for several of the documented differences between the effects of natural and rectangular light cycles on circadian entrainment. PMID- 15834111 TI - Importance of photoperiods in the regulation of ovarian activities in indian major carp Catla catla in an annual cycle. AB - The present study attempted for the first time to explore the importance of photoperiod in the regulation of seasonal ovarian functions in any subtropical major carp. Adult Indian major carp Catla catla were transferred to a long photoperiod (LP; LD 16:8) or a short photoperiod (SP; LD 8:16) for 30 days on 4 dates corresponding to the beginnings of 4 reproductive phases in an annual cycle, and responsiveness of the ovary was evaluated by comparison with the gonadal weight (I(G)), relative number of developing oocytes, serum levels of vitellogenin, and the activity of 2 important steroidogenic enzymes, that is, Delta(5)3beta-hydroxysteroid dehydrogenase and 17.beta-hydroxysteroid dehydrogenase, in the ovary of fish in a natural photoperiod. Exposure of fish to LP during the preparatory phase (February-March) resulted in a significant (p < 0.001) increase in the values of vitellogenin and in the activity of both the steroidogenic enzymes but not in the ovarian weight and in the relative number of different stages of oocytes. A more stimulatory influence of LP was noted during the prespawning phase (April-May), when precocious maturation of ovary was evident from a significant (p < 0.001) rise in the values of each studied features of ovarian functions. However, no ovarian response was found when the fish were transferred to LP during the spawning (July-August) and the postspawning (September-October) phases. On the other hand, the SP was found to have an inhibitory influence on ovarian growth and maturation during the prespawning and the spawning phases or to have no influences on ovarian functions during the preparatory and the postspawning phases of an annual cycle. The results of our study provide the first evidence that photoperiod per se plays an important role in the seasonal maturation of ovary in a subtropical freshwater major carp. PMID- 15834112 TI - Relative coordination to unknown "weak zeitgebers" in free-running blind individuals. AB - Light is the primary synchronizer of the human biological clock. In more than half of those blind individuals who completely lack light perception, the absence of photic input to the hypothalamic circadian pacemaker results in rhythms that free-run (blind free-runners [BFRs]) with a period typically greater than 24 h. The remainder are entrained, although sometimes at an abnormal phase angle. It is presumed that weak as-yet-to-be-identified time cues provide the necessary resetting stimulus in these entrained individuals. These weak zeitgebers might be expected to modulate the observed circadian period in blind people who are not actually entrained by them. The authors report here the results from 5 BFRs (average linear regression period +/-SD of 24.31 +/- 0.06 h) who had high resolution (many and frequent) phase assessments. All 5 subjects demonstrated a similar and reproducible pattern of changes in observed period (period response curves) indicative of relative coordination. The precise shape of the period response curve to weak zeitgebers has implications for the entrainment of BFRs using exogenous melatonin administration or other nonphotic stimuli. Sighted individuals may also be affected by such weak zeitgebers, which may be obscured by the stronger light/dark cycle. PMID- 15834114 TI - Stability of melatonin and temperature as circadian phase markers and their relation to sleep times in humans. AB - Circadian rhythms of core body temperature and melatonin are commonly used as phase markers of the circadian clock. Melatonin is a more stable marker of circadian phase when measured under constant routine conditions. However, little is known about the variability of these phase markers under less controlled conditions. Moreover, there is little consensus about the preferred method of analysis. The objective of this study was to assess various methods of calculating melatonin and temperature phase in subjects with regular sleep schedules living in their natural environment. Baseline data were analyzed from 42 healthy young subjects who were studied on at least two occasions. Each hospital admission was separated by at least 3 weeks. Subjects were instructed to maintain a regular sleep schedule, which was monitored for 1 week before admission by sleep logs and actigraphy. Subjects spent one habituation night under controlled conditions prior to collecting baseline temperature and melatonin measurements. The phase of the melatonin rhythm was assessed by 9 different methods. The temperature nadir (Tmin) was estimated using both Cleveland and Cosine curve fitting procedures, with and without demasking. Variability between admissions was assessed by correlation analysis and by the mean absolute difference in timing of the phase estimates. The relationship to sleep times was assessed by correlation of sleep onset or sleep offset with the various phase markers. Melatonin phase markers were more stable and more highly correlated with the timing of sleep than estimates of Tmin. Of the methods for estimating Tmin, simple cosine analysis was the least variable. In addition, sleep offset was more strongly correlated with the various phase markers than sleep onset. The relative measures of melatonin offset had the highest correlation coefficients, the lowest study-to-study variability, and were more strongly associated with sleep timing than melatonin onsets. Concordance of the methods of analysis suggests a tendency for the declining phase of the melatonin profile to be more stable and reliable than either markers of melatonin onset or measures of the termination of melatonin synthesis. PMID- 15834113 TI - Intrinsic period and light intensity determine the phase relationship between melatonin and sleep in humans. AB - The internal circadian clock and sleep-wake homeostasis regulate the timing of human brain function, physiology, and behavior so that wakefulness and its associated functions are optimal during the solar day and that sleep and its related functions are optimal at night. The maintenance of a normal phase relationship between the internal circadian clock, sleep-wake homeostasis, and the light-dark cycle is crucial for optimal neurobehavioral and physiological function. Here, the authors show that the phase relationship between these factors-the phase angle of entrainment (psi)-is strongly determined by the intrinsic period (tau) of the master circadian clock and the strength of the circadian synchronizer. Melatonin was used as a marker of internal biological time, and circadian period was estimated during a forced desynchrony protocol. The authors observed relationships between the phase angle of entrainment and intrinsic period after exposure to scheduled habitual wakefulness-sleep light dark cycle conditions inside and outside of the laboratory. Individuals with shorter circadian periods initiated sleep and awakened at a later biological time than did individuals with longer circadian periods. The authors also observed that light exposure history influenced the phase angle of entrainment such that phase angle was shorter following exposure to a moderate bright light (approximately 450 lux)-dark/wakefulness-sleep schedule for 5 days than exposure to the equivalent of an indoor daytime light (approximately 150 lux) dark/wakefulness-sleep schedule for 2 days. These findings demonstrate that neurobiological and environmental factors interact to regulate the phase angle of entrainment in humans. This finding has important implications for understanding physiological organization by the brain's master circadian clock and may have implications for understanding mechanisms underlying circadian sleep disorders. PMID- 15834115 TI - You whining at me? PMID- 15834118 TI - Adiponectin promotes adipocyte differentiation, insulin sensitivity, and lipid accumulation. AB - Adiponectin is secreted from adipocytes, and low circulating levels have been epidemiologically associated with obesity, insulin resistance, type 2 diabetes, and cardiovascular disease. To investigate whether adiponectin could exert autocrine effects in adipocytes, we expressed the adiponectin gene in 3T3-L1 fibroblasts. We observed that 3T3-L1 fibroblasts expressing adiponectin have a fast growth phase and reach confluence more rapidly compared with control cells or LacZ-transduced cells. Furthermore, cells with overexpressed adiponectin were observed to differentiate into adipocytes more rapidly, and during adipogenesis, they exhibited more prolonged and robust gene expression for related transcriptional factors, CCAAT/enhancer binding protein alpha (C/EBP2), peroxisome proliferator-activated receptor gamma (PPARgamma), and adipocyte determination and differentiation factor 1/sterol-regulatory element binding protein 1c (ADD1/SREBP1c) and earlier suppression of PPARgamma coactivator-1alpha (PGC-1alpha). In fully differentiated adipocytes, adiponectin-overexpressing cells accumulated more and larger lipid droplets compared with control cells. Also, adiponectin increased insulin's ability to maximally stimulate glucose uptake by 78% through increased glucose transporter 4 (GLUT4) gene expression and increased GLUT4 recruitment to the plasma membrane. These data suggest a new role for adiponectin as an autocrine factor in adipose tissues: promoting cell proliferation and differentiation from preadipocytes into adipocytes, augmenting programmed gene expression responsible for adipogenesis, and increasing lipid content and insulin responsiveness of the glucose transport system in adipocytes. PMID- 15834119 TI - A simple and rapid method to measure cholesterol binding to P450s and other proteins. AB - Cholesterol plays an important role in cellular function and membrane compartmentalization and is involved in the interaction with more than a dozen of different proteins. Using three cholesterol-metabolizing cytochrome P450s (P450s 7A1, 46A1, and 11A1), we have developed a rapid and simple assay for measurements of nanomolar to micromolar cholesterol affinities. In this assay, the P450 is incubated with a fixed amount of radiolabeled cholesterol and varying concentrations of cold cholesterol followed by separation of free and protein bound cholesterol via filtration through a membrane. Free cholesterol is found in the flow-through fraction, whereas P450 binds to the membrane. The radioactivity of the membranes is then measured, and a saturation curve is generated after correction for nonspecific binding of cholesterol to the filter. The validity of the filter assay was confirmed by spectral assay, a traditional method to evaluate the interaction of the P450 enzymes with their substrates. Two types of membranes, one binding positively charged proteins and another binding negatively charged proteins, were identified. These membranes were also found to hold proteins through hydrophobic interactions. Thus, the cholesterol binding properties of a wide variety of proteins could be characterized using this filter assay. PMID- 15834120 TI - Shotgun lipidomics of phosphoethanolamine-containing lipids in biological samples after one-step in situ derivatization. AB - This article presents a novel methodology for the analysis of ethanolamine glycerophospholipid (PE) and lysoPE molecular species directly from lipid extracts of biological samples. Through brief treatment of lipid extracts with fluorenylmethoxylcarbonyl (Fmoc) chloride, PE and lysoPE species were selectively derivatized to their corresponding carbamates. The reaction solution was infused directly into the ion source of an electrospray ionization mass spectrometer after appropriate dilution. The facile loss of the Fmoc moiety dramatically enhanced the analytic sensitivity and allowed the identification and quantitation of low-abundance molecular species. A detection limitation of attomoles (amoles) per microliter for PE and lysoPE analysis was readily achieved using this technique (at least a 100-fold improvement from our previous method) with a >15,000-fold dynamic range. Through intrasource separation and multidimensional mass spectrometry array analysis of derivatized species, marked improvements in signal-to-noise ratio, molecular species identification, and quantitation can be realized. The procedure is both simple and effective and can be extended to analyze many other lipid classes or other cellular metabolites by adjustments in specific derivatization conditions. Thus, through judicious derivatization, a new dimension exploiting specific functional reactivities in each lipid class can be used in conjunction with shotgun lipidomics to penetrate farther into the low abundance regime of cellular lipidomes. PMID- 15834121 TI - Thematic review series: The immune system and atherogenesis. Cytokines affecting endothelial and smooth muscle cells in vascular disease. AB - The cellular and extracellular matrix accumulations that comprise the lesions of atherosclerosis are driven by local release of cytokines at sites of predilection for lesion formation, and by the specific attraction and activation of cells expressing receptors for these cytokines. Although cytokines were originally characterized for their potent effects on immune and inflammatory cells, they also promote endothelial cell dysfunction and alter smooth muscle cell (SMC) phenotype and function, which can contribute to or retard vascular pathologies. This review summarizes in vivo studies that have characterized endothelial- and smooth muscle-specific effects of altering cytokine signaling in vascular disease. Although multiple reports have identified cytokines as pivotal players in endothelial and SMC responses in vascular disease, they also have highlighted the need to delineate the critical genes and specific cellular functions regulated by individual cytokine signaling pathways. PMID- 15834122 TI - Cholesterol-lowering effects of dietary blue lupin (Lupinus angustifolius L.) in intact and ileorectal anastomosed pigs. AB - The present study was undertaken to investigate the effect of cholesterol enriched casein (CAS) and blue lupin seed (BL) diets on the cholesterol metabolism of intact (INT) and ileorectal anastomosed (IRA) pigs. For 3 weeks, four groups of six pigs were allocated to the treatments (CAS-INT, CAS-IRA, BL INT, and BL-IRA). Diet-induced hypercholesterolemia was inhibited by the BL through a substantial decrease in plasma LDL-cholesterol. The BL also reduced liver esterified and total cholesterol, increased hepatic LDL receptor synthesis and HMG-CoA reductase activity, and stimulated intestinal bile acid reabsorption. The neutral sterol output was higher in BL- than in CAS-fed pigs. The bile acid output was lower in IRA than in INT pigs. Surgery also prevented steroid microbial transformation, but it did not influence plasma cholesterol levels. These results suggest that the hypocholesterolemic effect of the BL, compared with the CAS, is attributable to impaired intestinal cholesterol absorption, probably involving increased bile acid reabsorption and higher contents of dietary phytosterols, both factors that reduce the micellar solubilization of cholesterol. Furthermore, according to our data, the contribution of the large intestine to cholesterol metabolism is very weak. PMID- 15834123 TI - Loss of G2A promotes macrophage accumulation in atherosclerotic lesions of low density lipoprotein receptor-deficient mice. AB - Lysophosphatidylcholine (LPC) is considered a major proatherogenic component of oxidized low density lipoprotein based on its proinflammatory actions in vitro. LPC stimulates macrophage and T-cell chemotaxis via the G protein-coupled receptor G2A and may thus promote inflammatory cell infiltration during atherosclerotic lesion development. However, G2A also mediates proapoptotic effects of LPC and may therefore promote the death of inflammatory cells within developing lesions. To determine how these effects of LPC modify atherogenesis, we examined atherosclerotic lesion development in G2A-sufficient and G2A deficient low density lipoprotein receptor knockout mice. Although LPC species capable of activating G2A-dependent responses were increased during lesion development, G2A-deficient mice developed lesions similar in size to those in their G2A-sufficient counterparts. Loss of G2A during atherosclerotic lesion development did not reduce macrophage and T-cell infiltration but instead resulted in increased lesional macrophage content associated with reduced numbers of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeled cells and decreased collagen deposition. These data indicate that the ability of LPC to stimulate macrophage and T-cell chemotaxis via G2A is not manifested in vivo and that G2A-mediated proapoptotic rather than chemotactic action is most penetrant during atherogenesis and may modify the stability of atherosclerotic lesions by promoting macrophage death. PMID- 15834124 TI - Lipid imaging by gold cluster time-of-flight secondary ion mass spectrometry: application to Duchenne muscular dystrophy. AB - Imaging with time-of-flight secondary ion mass spectrometry (TOF-SIMS) has expanded very rapidly with the development of gold cluster ion sources (Au(3+)). It is now possible to acquire ion density maps (ion images) on a tissue section without any treatment and with a lateral resolution of few micrometers. In this article, we have taken advantage of this technique to study the degeneration/regeneration process in muscles of a Duchenne muscular dystrophy model mouse. Specific distribution of different lipid classes (fatty acids, triglycerides, phospholipids, tocopherol, coenzyme Q9, and cholesterol) allows us to distinguish three different regions on a mouse leg section: one is destroyed, another is degenerating (oxidative stress and deregulation of the phosphoinositol cycle), and the last one is stable. TOF-SIMS imaging shows the ability to localize directly on a tissue section a great number of lipid compounds that reflect the state of the cellular metabolism. PMID- 15834125 TI - Endothelial lipase releases saturated and unsaturated fatty acids of high density lipoprotein phosphatidylcholine. AB - We assessed the ability of endothelial lipase (EL) to hydrolyze the sn-1 and sn-2 fatty acids (FAs) from HDL phosphatidylcholine. For this purpose, reconstituted discoidal HDLs (rHDLs) that contained free cholesterol, apolipoprotein A-I, and either 1-palmitoyl-2-oleoylphosphatidylcholine, 1-palmitoyl-2 linoleoylphosphatidylcholine, or 1-palmitoyl-2-arachidonylphosphatidylcholine were incubated with EL- and control (LacZ)-conditioned media. Gas chromatography analysis of the reaction mixtures revealed that both the sn-1 (16:0) and sn-2 (18:1, 18:2, and 20:4) FAs were liberated by EL. The higher rate of sn-1 FA cleavage compared with sn-2 FA release generated corresponding sn-2 acyl lyso species as determined by MS analysis. EL failed to release sn-2 FA from rHDLs containing 1-O-1'-hexadecenyl-2-arachidonoylphosphatidylcholine, whose sn-1 position contained a nonhydrolyzable alkyl ether linkage. The lack of phospholipase A(2) activity of EL and its ability to liberate [(14)C]FA from [(14)C]lysophosphatidylcholine (lyso-PC) led us to conclude that EL-mediated deacylation of phosphatidylcholine (PC) is initiated at the sn-1 position, followed by the release of the remaining FA from the lyso-PC intermediate. Thin layer chromatography analysis of cellular lipids obtained from EL-overexpressing cells revealed a pronounced accumulation of [(14)C]phospholipid and [(14)C]triglyceride upon incubation with 1-palmitoyl-2-[1-(14)C]linoleoyl-PC labeled HDL(3), indicating the ability of EL to supply cells with unsaturated FAs. PMID- 15834126 TI - The triacylglycerol synthesis enzyme DGAT1 also catalyzes the synthesis of diacylglycerols, waxes, and retinyl esters. AB - The final step of triacylglycerol biosynthesis is catalyzed by acyl CoA:diacylglycerol acyltransferase (DGAT) enzymes. The two known DGATs, DGAT1 and DGAT2, are encoded by unrelated genes. Although both DGAT1 and DGAT2 knockout mice have reduced tissue triacylglycerol contents, they have disparate phenotypes, prompting us to investigate whether the two enzymes have unrecognized functional differences. We now report that DGAT1 exhibits additional acyltransferase activities in vitro, including those of acyl CoA:monoacylglycerol acyltransferase (MGAT), wax monoester and wax diester synthases, and acyl CoA:retinol acyltransferase (ARAT), which catalyze the synthesis of diacylglycerols, wax esters, and retinyl esters, respectively. These activities were demonstrated in in vitro assays with membranes from insect cells or homogenates from COS7 cells overexpressing DGAT1. Wax synthase and ARAT activities were also demonstrated in intact COS7 cells expressing DGAT1. Additionally, cells and tissues from DGAT1-deficient mice exhibited reduced ARAT activity, and the mice had increased levels of unesterified retinol in their livers on a high-retinol diet. Our findings indicate that DGAT1 can utilize a variety of acyl acceptors as substrates in vitro and suggest that these activities may be relevant to the in vivo functions of DGAT1. PMID- 15834127 TI - Evidence for multiple complementary pathways for efficient cholesterol absorption in mice. AB - Apolipoprotein B (apoB)-dependent and apoB-independent pathways for cholesterol transport have been described in cultured cells. Here, we show that the apoB independent pathway involves apoA-I-containing high density lipoproteins (HDLs). Cholesterol secretion by the HDLs, but not by the apoB pathway, was significantly reduced in primary enterocytes isolated from chow- and cholesterol-fed apoA-I(-/ ) mice. These enterocytes were capable of cholesterol efflux when apoA-I was provided extracellularly. In apoA-I(-/-) mice, the absorption of a bolus of cholesterol was similar in control and apoA-I(-/-) mice fed chow or high cholesterol diet. However, short-term studies revealed that cholesterol absorption was occurring over longer lengths of the intestine, and cholesterol but not triglyceride transport to the plasma and liver in chow- and cholesterol fed apoA-I(-/-) mice was significantly reduced. These studies indicate that in apoA-I deficiency, there is a delay in cholesterol absorption, but cholesterol is eventually absorbed because of the compensatory apoB pathway. Nonetheless, long term studies involving multiple feedings showed significant reduction in cholesterol absorption after 4 days. We propose that multiple compensatory mechanisms ensure efficient cholesterol absorption in mice. PMID- 15834128 TI - Compartmental modeling to quantify alpha-linolenic acid conversion after longer term intake of multiple tracer boluses. AB - To estimate in vivo alpha-linolenic acid (ALA; C18:3n-3) conversion, 29 healthy subjects consumed for 28 days a diet providing 7% of energy from linoleic acid (C18:2n-6) and 0.4% from ALA. On day 19, subjects received a single bolus of 30 mg of uniformly labeled [(13)C]ALA and for the next 8 days 10 mg twice daily. Fasting plasma phospholipid concentrations of (12)C- and (13)C-labeled ALA, eicosapentaenoic acid (EPA; C20:5n-3), docosapentaenoic acid (DPA; C22:5n-3), and docosahexaenoic acid (DHA; C22:6n-3) were determined on days 19, 21, 23, 26, 27, and 28. To estimate hepatic conversion of n-3 fatty acids, a tracer model was developed based on the averaged (13)C data of the participants. A similar tracee model was solved using the averaged (12)C values, the kinetic parameters derived from the tracer model, and mean ALA consumption. ALA incorporation into plasma phospholipids was estimated by solving both models simultaneously. It was found that nearly 7% of dietary ALA was incorporated into plasma phospholipids. From this pool, 99.8% was converted into EPA and 1% was converted into DPA and subsequently into DHA. The limited incorporation of dietary ALA into the hepatic phospholipid pool contributes to the low hepatic conversion of ALA into EPA. A low conversion of ALA-derived EPA into DPA might be an additional obstacle for DHA synthesis. PMID- 15834129 TI - Determinants of variation in serum paraoxonase enzyme activity in baboons. AB - Paraoxonase (PON), an HDL-associated enzyme, is one of many circulating antioxidants thought to play a vital protective role. To better understand the determinants of quantitative variation in serum PON activity, we assayed PON in samples from 611 pedigreed baboons fed three diets. PON was measured enzymatically; the main determinant of variation was genetic and consisted of at least three components: two loci detected by linkage analyses and a residual polygenic component. Multipoint linkage analyses gave peak log of the odds (LOD) scores on the baboon homolog of human chromosome 7q21-22 (near PON1, the structural gene) of 9.1 on the low-cholesterol, high-fat diet and 4.1 on the high cholesterol, high-fat diet (genome-wide P values were 1 x 10(-8) and 0.0018, respectively). Surprisingly, a second locus on the baboon homolog of human chromosome 12q13 gave a LOD score of 2.9 on the high-cholesterol, high-fat diet (genome-wide P value was 0.032). We identified several significant covariates, including age, sex, diet, and apolipoprotein A-I concentrations. We estimate that 53% of total trait variation in baboons is explained by genes and 17% by covariates, thus accounting for approximately 70% of total variation in baboon PON. Although the generation of free radicals is influenced primarily by environmental factors, our findings suggest strong genetic regulation of one component in the antioxidant defense system that plays a major role in susceptibility to atherosclerosis. PMID- 15834130 TI - Membrane microdomains in hepatocytes: potential target areas for proteins involved in canalicular bile secretion. AB - The formation of hepatic bile requires that water be transported across liver epithelia. Rat hepatocytes express three aquaporins (AQPs): AQP8, AQP9, and AQP0. Recognizing that cholesterol and sphingolipids are thought to promote the assembly of proteins into specialized membrane microdomains, we hypothesized that canalicular bile secretion involves the trafficking of vesicles to and from localized lipid-enriched microdomains in the canalicular plasma membrane. Hepatocyte plasma membranes were sonicated in Triton and centrifuged overnight on a sucrose gradient to yield a Triton-soluble pellet and a Triton-insoluble, sphingolipid-enriched microdomain fraction at the 5%/30% sucrose interface. The detergent-insoluble portion of the hepatocyte plasma membrane was enriched in alkaline phosphatase (a microdomain-positive marker) and devoid of amino peptidase N (a microdomain-negative marker), enriched in caveolin, both AQP8 and AQP9, but negative for clathrin. The microdomain fractions contained chloride bicarbonate anion exchanger isoform 2 and multidrug resistance-associated protein 2. Exposure of isolated hepatocytes to glucagon increased the expression of AQP8 but not AQP9 in the microdomain fractions. Sphingolipid analysis of the insoluble fraction showed the predominant species to be sphingomyelin. These data support the presence of sphingolipid-enriched microdomains of the hepatocyte membrane that represent potential localized target areas for the clustering of AQPs and functionally related proteins involved in canalicular bile secretion. PMID- 15834131 TI - Alternative pre-mRNA splicing governs expression of a conserved acidic transactivation domain in myocyte enhancer factor 2 factors of striated muscle and brain. AB - Myocyte enhancer factor 2 (MEF2) transcription factors play pivotal roles in striated muscle, neuron, and lymphocyte gene expression and are targets of stress and calcium-mediated signaling. All MEF2 gene products have a common DNA binding and dimerization domain, but MEF2 transcripts are alternatively spliced among coding exons to produce splicing isoforms. In vertebrate MEF2A, -C, and -D, a splice versus no-splice option gives forms that include or exclude a short domain that we designate beta. We show that mRNAs containing beta are expressed predominantly in striated muscle and brain and that splicing to include beta is induced during myocyte differentiation. MEF2 beta+ isoforms are more robust than beta- forms in activating MEF2-responsive reporters despite similar expression levels. One-hybrid transcription assays using Gal4-MEF2 fusions show similar distinctions in the transactivation produced by beta+ versus beta- isoforms in all cell types tested, including myocytes. beta function is position-independent and exists in all MEF2 splicing variant contexts. The activity is not due to cis effects on MEF2 DNA binding or dimerization nor are established transcription factor or coactivator interactions involved. Each MEF2 beta domain contains multiple acidic residues, mutation of which abolishes function. Despite a location between the p38 MAPK docking domain and Thr phosphoacceptors of MEF2A and MEF2C, inclusion of beta does not influence responses of these factors to this signaling pathway. Thus, a conserved pattern of alternative splicing in vertebrate MEF2 genes generates an acidic activation domain in MEF2 proteins selectively in tissues where MEF2 target genes are highly expressed. PMID- 15834132 TI - Convergence of cell cycle regulation and growth factor signals on GRASP65. AB - Together with other Golgi matrix components, GRASP65 contributes to the stacking of Golgi cisternae in interphase cells. During mitosis, GRASP65 is heavily phosphorylated, and in turn, cisternal stacking is inhibited leading to the breakdown of the Golgi apparatus. Here we show that GRASP65 is phosphorylated on serine 277 in interphase cells, and this is strongly enhanced in response to the addition of serum or epidermal growth factor. This is directly mediated by ERK suggesting that GRASP65 has some role in growth factor signal transduction. Phosphorylation of Ser-277 is also dramatically increased during mitosis, however this is mediated by Cdk1 and not by ERK. The microinjection of recombinant GRASP65 without N-terminal myristoylation or a peptide fragment containing Ser 277 into the cytosol of normal rat kidney cells inhibits passage through mitosis. This effect is abolished when Ser-277 is replaced with alanine suggesting the phosphorylation of Ser-277 plays an important role in cell cycle regulation. The convergence of cell cycle regulation and growth factor signals on GRASP65 Ser-277 suggests that GRASP65 may function as a signal integrator controlling the cell growth. PMID- 15834133 TI - Substrate-specific modulation of a multisubstrate proteinase. C-terminal processing of fibrillar procollagens is the only BMP-1-dependent activity to be enhanced by PCPE-1. AB - Members of the bone morphogenetic protein-1/tolloid (BMP-1/Tld) family of metalloproteinases, also known as procollagen C-proteinases (PCPs), control multiple biological events (including matrix assembly, cross-linking, cell adhesion/migration and pattern formation) through enzymatic processing of several extracellular substrates. PCP activities on fibrillar procollagens can be stimulated by another family of extracellular proteins, PCP enhancers (PCPE-1, PCPE-2), which lack intrinsic enzymatic activity. While PCPs have multiple substrates, the extent to which PCPEs is involved in the processing of proteins other than fibrillar procollagens is unknown. In the experiments reported here, PCPE-1 was found to have no effect on the in vitro BMP-1 processing of procollagen VII, the procollagen V N-propeptide, the laminin 5 gamma2 chain, osteoglycin, prolysyl oxidase, or chordin. In contrast, PCPE-1 enhanced C terminal processing of human fibrillar procollagen III but only when this substrate was in its native, disulfide-bonded conformation. Surprisingly, processing of procollagen III continued to be enhanced when essentially all the triple-helical region was removed. These and previous results (Ricard-Blum, S., Bernocco, S., Font, B., Moali, C., Eichenberger, D., Farjanel, J., Burchardt, E. R., van der Rest, M., Kessler, E., and Hulmes, D. J. S. (2002) J. Biol. Chem. 277, 33864-33869; Bernocco, S., Steiglitz, B. M., Svergun, D. I., Petoukhov, M. V., Ruggiero, F., Ricard-Blum, S., Ebel, C., Geourjon, C., Deleage, G., Font, B., Eichenberger, D., Greenspan, D. S., and Hulmes, D. J. S. (2003) J. Biol. Chem. 278, 7199-7205) indicate that the mechanism of PCPE-1 action involves recognition sites in both the C-propeptide domain and in the C-telopeptide region of the procollagen molecule. PCPEs therefore define a new class of extracellular adaptor proteins that stimulate proteinase activity in a substrate-specific manner, thereby providing a new target for the selective regulation of PCP activity on fibrillar procollagen substrates. PMID- 15834134 TI - Prostaglandin E2 receptors EP2 and EP4 are down-regulated during differentiation of mouse osteoclasts from their precursors. AB - Prostaglandin E2 (PGE2) has been proposed to be a potent stimulator of bone resorption. However, PGE2 itself has been shown to directly inhibit bone resorbing activity of osteoclasts. We examined the role of PGE2 in the function of mouse osteoclasts formed in vitro. Bone marrow macrophage osteoclast precursors expressed PGE2 receptors EP1, EP2, EP3beta, and EP4, and the expression of EP2 and EP4 was down-regulated during osteoclastic differentiation induced by receptor activator of NF-kappaB ligand and macrophage colony stimulating factor. In contrast, functional EP1 was continuously expressed in mature osteoclasts. PGE2 as well as calcitonin caused intracellular Ca2+ influx in osteoclasts. However, PGE2 and 17-phenyltrinol-PGE2 (an EP1 agonist) failed to inhibit actin-ring formation and pit formation by osteoclasts cultured on dentine slices. When EP4 was expressed in osteoclasts using an adenovirus carrying EP4 cDNA, both actin-ring and pit-forming activities of osteoclasts were inhibited in an infectious unit-dependent manner. Treatment of EP4-expressing osteoclasts with PGE2 further inhibited their actin-ring and pit-forming activities. Such inhibitory effects of EP4-mediated signals on osteoclast function are similar to those that are calcitonin receptor-mediated. Thus, osteoclast precursors down regulate their own EP2 and EP4 levels during their differentiation into osteoclasts to escape inhibitory effects of PGE2 on bone resorption. PMID- 15834135 TI - Induction of KLF2 by fluid shear stress requires a novel promoter element activated by a phosphatidylinositol 3-kinase-dependent chromatin-remodeling pathway. AB - Fluid shear stress maintains vascular homeostasis by influencing endothelial gene expression. One mechanism by which shear stress achieves this is through the induction of transcription factors including Kruppel-like factor 2(KLF2). We have previously reported that a 62-bp region of the KLF2 promoter is responsible for its shear stress-induced expression via the binding of nuclear factors. In this study, we find that the 62-bp shear stress response region contains a 30-bp tripartite palindrome motif. Electrophoretic mobility supershift and chromatin immunoprecipitation assays demonstrate that PCAF (P-300/cAMP-response element binding protein-binding protein-associated factor)) and heterogeneous nuclear ribonucleoprotein D bind this region as components of the shear stress regulatory complex. We have also characterized a PI3K-dependent/Akt-independent pathway responsible for shear stress-induced KLF2 nuclear binding, promoter activation, and mRNA expression. Furthermore, the shear stress response region of the KLF2 promoter was specifically immunoprecipitated by antibodies against acetylated histones H3 and H4 in shear-stressed but not static hemangioendothelioma cells. The acetylation of these histones was blocked by PI3K inhibition. Finally, we have found that KLF2 increases endothelial nitric-oxide synthase expression in murine endothelial cultures, an effect that is also blocked by PI3K inhibition. These results define the DNA regulatory element, signal transduction pathway, and molecular mechanism activating the flow-dependent expression of a vital endothelial transcription factor. PMID- 15834136 TI - Expression profile of osteoblast lineage at defined stages of differentiation. AB - The inherent heterogeneity of bone cells complicates the interpretation of microarray studies designed to identify genes highly associated with osteoblast differentiation. To overcome this problem, we have utilized Col1a1 promoter-green fluorescent protein transgenic mouse lines to isolate bone cells at distinct stages of osteoprogenitor maturation. Comparison of gene expression patterns from unsorted or isolated sorted bone cell populations at days 7 and 17 of calvarial cultures revealed an increased specificity regarding which genes are selectively expressed in a subset of bone cell types during differentiation. Furthermore, distinctly different patterns of gene expression associated with major signaling pathways (Igf1, Bmp, and Wnt) were observed at different levels of maturation. Some of our data differ from current models of osteoprogenitor cell differentiation and emphasize components of the pathways that were not revealed in studies based on a total cell population. Thus, applying methods to generate more homogeneous populations of cells at a defined level of cellular differentiation from a primary osteogenic culture is feasible and leads to a novel interpretation of the gene expression associated with increasing levels of osteoprogenitor maturation. PMID- 15834137 TI - DNA rearrangement in orthologous orp regions of the maize, rice and sorghum genomes. AB - The homeologous Orp1 and Orp2 regions of maize and the orthologous regions in sorghum and rice were compared by generating sequence data for >486 kb of genomic DNA. At least three genic rearrangements differentiate the maize Orp1 and Orp2 segments, including an insertion of a single gene and two deletions that removed one gene each, while no genic rearrangements were detected in the maize Orp2 region relative to sorghum. Extended comparison of the orthologous Orp regions of sorghum and japonica rice uncovered numerous genic rearrangements and the presence of a transposon-rich region in rice. Only 11 of 27 genes (40%) are arranged in the same order and orientation between sorghum and rice. Of the 8 genes that are uniquely present in the sorghum region, 4 were found to have single-copy homologs in both rice and Arabidopsis, but none of these genes are located near each other, indicating frequent gene movement. Further comparison of the Orp segments from two rice subspecies, japonica and indica, revealed that the transposon-rich region is both an ancient and current hotspot for retrotransposon accumulation and genic rearrangement. We also identify unequal gene conversion as a mechanism for maize retrotransposon rearrangement. PMID- 15834138 TI - A screen for genes regulating the wingless gradient in Drosophila embryos. AB - During the development of the Drosophila embryonic epidermis, the secreted Wingless protein initially spreads symmetrically from its source. At later stages, Wingless becomes asymmetrically distributed in a Hedgehog-dependent manner, to control the patterning of the embryonic epidermis. When Wingless is misexpressed in engrailed cells in hedgehog heterozygous mutant embryos, larvae show a dominant phenotype consisting of patches of naked cuticle in denticle belts. This dose-sensitive phenotype is a direct consequence of a change in Wg protein distribution. We used this phenotype to carry out a screen for identifying genes regulating Wingless distribution or transport in the embryonic epidermis. Using a third chromosome deficiency collection, we found several genomic regions that showed a dominant interaction. After using a secondary screen to test for mutants and smaller deficiencies, we identified three interacting genes: dally, notum, and brahma. We confirmed that dally, as well as its homolog dally-like, and notum affect Wingless distribution in the embryonic epidermis, directly or indirectly. Thus, our assay can be used effectively to screen for genes regulating Wingless distribution or transport. PMID- 15834139 TI - Accelerated evolution of the pituitary adenylate cyclase-activating polypeptide precursor gene during human origin. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide abundantly expressed in the central nervous system and involved in regulating neurogenesis and neuronal signal transduction. The amino acid sequence of PACAP is extremely conserved across vertebrate species, indicating a strong functional constraint during the course of evolution. However, through comparative sequence analysis, we demonstrated that the PACAP precursor gene underwent an accelerated evolution in the human lineage since the divergence from chimpanzees, and the amino acid substitution rate in humans is at least seven times faster than that in other mammal species resulting from strong Darwinian positive selection. Eleven human-specific amino acid changes were identified in the PACAP precursors, which are conserved from murine to African apes. Protein structural analysis suggested that a putative novel neuropeptide might have originated during human evolution and functioned in the human brain. Our data suggested that the PACAP precursor gene underwent adaptive changes during human origin and may have contributed to the formation of human cognition. PMID- 15834140 TI - Spontaneous mutational correlations for life-history, morphological and behavioral characters in Caenorhabditis elegans. AB - The pattern of mutational covariance among traits plays a central, but largely untested, role in many theories in evolutionary genetics. Here we estimate the pattern of phenotypic, environmental, and mutational correlations for a set of life-history, behavioral, and morphological traits using 67 self-fertilizing lines of Caenorhabditis elegans, each having independently experienced an average of 370 generations of spontaneous mutation accumulation. Bivariate relationships of mutational effects indicate the existence of extensive pleiotropy. We find that mutations may tend to produce manifold effects on suites of functionally related traits; however, our data do not support the idea of completely parcelated pleiotropy, in which functional units are separately affected by mutations. Positive net phenotypic and mutational correlations are common for life-history traits, with environmental correlations being comparatively smaller and of the same sign for most pairs of traits. Observed mutational correlations are shown to be higher than those produced by the chance accumulation of nonpleiotropic mutations in the same lines. PMID- 15834141 TI - Behavioral degradation under mutation accumulation in Caenorhabditis elegans. AB - Spontaneous mutations play a fundamental role in the maintenance of genetic variation in natural populations, the nature of inbreeding depression, the evolution of sexual reproduction, and the conservation of endangered species. Using long-term mutation-accumulation lines of the nematode Caenorhabditis elegans, we estimate the rate and magnitude of mutational effects for a suite of behaviors characterizing individual chemosensory responses to a repellant stimulus. In accordance with evidence that the vast majority of mutations are deleterious, we find that behavioral responses degrade over time as a result of spontaneous mutation accumulation. The rate of mutation for behavioral traits is roughly of the same order or slightly smaller than those previously estimated for reproductive traits and the average size of the mutational effects is also comparable. These results have important implications for the maintenance of genetic variation for behavior in natural populations as well as for expectations for behavioral change within endangered species and captive populations. PMID- 15834142 TI - A screen for genes that influence fibroblast growth factor signal transduction in Drosophila. AB - The misexpression of an activated form of the FGF receptor (FGFR) Breathless in conjunction with downstream-of-FGF-receptor (Dof), an essential signaling molecule of the FGF pathway, in the Drosophila eye imaginal discs impairs eye development and results in a rough eye phenotype. We used this phenotype in a gain-of-function screen to search for modifiers of FGF signaling. We identified 50 EP stocks with insertions defining at least 35 genes that affect the rough eye phenotype. Among these genes, 4 appear to be specific for FGFR signaling, but most of the genes also influence other signaling pathways, as assessed by their effects on rough eyes induced by other activated receptor tyrosine kinases (RTKs). Analysis of loss-of-function alleles of a number of these genes in embryos indicates that in many cases the products are provided maternally and are involved in germ cell development. At least two of the genes, sar1 and robo2, show a genetic interaction with a hypomorphic dof allele, suggesting that they participate in FGF-mediated morphogenetic events during embryogenesis. PMID- 15834143 TI - An efficient Monte Carlo method for estimating Ne from temporally spaced samples using a coalescent-based likelihood. AB - This article presents an efficient importance-sampling method for computing the likelihood of the effective size of a population under the coalescent model of Berthier et al. Previous computational approaches, using Markov chain Monte Carlo, required many minutes to several hours to analyze small data sets. The approach presented here is orders of magnitude faster and can provide an approximation to the likelihood curve, even for large data sets, in a matter of seconds. Additionally, confidence intervals on the estimated likelihood curve provide a useful estimate of the Monte Carlo error. Simulations show the importance sampling to be stable across a wide range of scenarios and show that the N(e) estimator itself performs well. Further simulations show that the 95% confidence intervals around the N(e) estimate are accurate. User-friendly software implementing the algorithm for Mac, Windows, and Unix/Linux is available for download. Applications of this computational framework to other problems are discussed. PMID- 15834144 TI - Survival analysis of life span quantitative trait loci in Drosophila melanogaster. AB - We used quantitative trait loci (QTL) mapping to evaluate the age specificity of naturally segregating alleles affecting life span. Estimates of age-specific mortality rates were obtained from observing 51,778 mated males and females from a panel of 144 recombinant inbred lines (RILs). Twenty-five QTL were found, having 80 significant effects on life span and weekly mortality rates. Generation of RILs from heterozygous parents enabled us to contrast effects of QTL alleles with the means of RIL populations. Most of the low-frequency alleles increased mortality, especially at younger ages. Two QTL had negatively correlated effects on mortality at different ages, while the remainder were positively correlated. Chromosomal positions of QTL were roughly concordant with estimates from other mapping populations. Our findings are broadly consistent with a mix of transient deleterious mutations and a few polymorphisms maintained by balancing selection, which together contribute to standing genetic variation in life span. PMID- 15834146 TI - Toward a neutral evolutionary model of gene expression. AB - We introduce a stochastic model that describes neutral changes of gene expression over evolutionary time as a compound Poisson process where evolutionary events cause changes of expression level according to a given probability distribution. The model produces simple estimators for model parameters and allows discrimination between symmetric and asymmetric distributions of evolutionary expression changes along an evolutionary lineage. Furthermore, we introduce two measures, the skewness of expression difference distributions and relative difference of evolutionary branch lengths, which are used to quantify deviation from clock-like behavior of gene expression distances. Model-based analyses of gene expression profiles in primate liver and brain samples yield the following results: (1) The majority of gene expression changes are consistent with a neutral model of evolution; (2) along evolutionary lineages, upward changes in expression are less frequent but of greater average magnitude than downward changes; and (3) the skewness measure and the relative branch length difference confirm that an acceleration of gene expression evolution occurred on the human lineage in brain but not in liver. We discuss the latter result with respect to a neutral model of transcriptome evolution and show that a small number of genes expressed in brain can account for the observed data. PMID- 15834145 TI - The epsilon-subunit of mitochondrial ATP synthase is required for normal spindle orientation during the Drosophila embryonic divisions. AB - We describe the maternal-effect and zygotic phenotypes of null mutations in the Drosophila gene for the epsilon-subunit of mitochondrial ATP synthase, stunted (sun). Loss of zygotic sun expression leads to a dramatic delay in the growth rate of first instar larvae and ultimately death. Embryos lacking maternally supplied sun (sun embryos) have a sixfold reduction in ATP synthase activity. Cellular analysis of sun embryos shows defects only after the nuclei have migrated to the cortex. During the cortical divisions the actin-based metaphase and cellularization furrows do not form properly, and the nuclei show abnormal spacing and division failures. The most striking abnormality is that nuclei and spindles form lines and clusters, instead of adopting a regular spacing. This is reflected in a failure to properly position neighboring nonsister centrosomes during the telophase-to-interphase transition of the cortical divisions. Our study is consistent with a role for Sun in mitochondrial ATP synthesis and suggests that reduced ATP levels selectively affect molecular motors. As Sun has been identified as the ligand for the Methuselah receptor that regulates aging, Sun may function both within and outside mitochondria. PMID- 15834147 TI - Evolutionary history of Silene latifolia sex chromosomes revealed by genetic mapping of four genes. AB - The sex chromosomes of dioecious white campion, Silene latifolia (Caryophyllaceae), are of relatively recent origin (10-20 million years), providing a unique opportunity to trace the origin and evolution of sex chromosomes in this genus by comparing closely related Silene species with and without sex chromosomes. Here I demonstrate that four genes that are X-linked in S. latifolia are also linked in nondioecious S. vulgaris, which is consistent with Ohno's (1967) hypothesis that sex chromosomes evolve from a single pair of autosomes. I also report a genetic map for four S. latifolia X-linked genes, SlX1, DD44X, SlX4, and a new X-linked gene SlssX, which encodes spermidine synthase. The order of the genes on the S. latifolia X chromosome and divergence between the homologous X- and Y-linked copies of these genes supports the "evolutionary strata" model, with at least three consecutive expansions of the nonrecombining region on the Y chromosome (NRY) in this plant species. PMID- 15834148 TI - A genetic screen for suppressors of Drosophila NSF2 neuromuscular junction overgrowth. AB - The Drosophila larval neuromuscular system serves as a valuable model for studying the genes required for synaptic development and function. N Ethylmaleimide sensitive factor (NSF) is a molecule known to be important in vesicular trafficking but neural expression of a dominant negative form of NSF2 induces an unexpected overgrowth of the Drosophila larval neuromuscular synapse. We have taken a genetic approach to understanding this novel phenotype by conducting a gain-of-function modifier screen to isolate genes that interact with the overgrowth phenotype. Our approach was to directly visualize the neuromuscular junction (NMJ) using a GFP transgene and screen for suppressors of NMJ overgrowth using the Gene Search collection of P-element insertions. Of the 3000 lines screened, we identified 99 lines that can partially restore the normal phenotype. Analysis of the GS element insertion sites by inverse PCR and comparison of the flanking DNA sequence to the Drosophila genome sequence revealed nearby genes for all but 10 of the 99 lines. The recovered genes, both known and predicted, include transcription factors, cytoskeletal elements, components of the ubiquitin pathway, and several signaling molecules. This collection of genes that suppress the NSF2 neuromuscular junction overgrowth phenotype is a valuable resource in our efforts to further understand the role of NSF at the synapse. PMID- 15834149 TI - Model selection in binary trait locus mapping. AB - Quantitative trait locus (QTL) mapping methodology for continuous normally distributed traits is the subject of much attention in the literature. Binary trait locus (BTL) mapping in experimental populations has received much less attention. A binary trait by definition has only two possible values, and the penetrance parameter is restricted to values between zero and one. Due to this restriction, the infinitesimal model appears to come into play even when only a few loci are involved, making selection of an appropriate genetic model in BTL mapping challenging. We present a probability model for an arbitrary number of BTL and demonstrate that, given adequate sample sizes, the power for detecting loci is high under a wide range of genetic models, including most epistatic models. A novel model selection strategy based upon the underlying genetic map is employed for choosing the genetic model. We propose selecting the "best" marker from each linkage group, regardless of significance. This reduces the model space so that an efficient search for epistatic loci can be conducted without invoking stepwise model selection. This procedure can identify unlinked epistatic BTL, demonstrated by our simulations and the reanalysis of Oncorhynchus mykiss experimental data. PMID- 15834152 TI - Effective size of a fluctuating age-structured population. AB - Previous theories on the effective size of age-structured populations assumed a constant environment and, usually, a constant population size and age structure. We derive formulas for the variance effective size of populations subject to fluctuations in age structure and total population size produced by a combination of demographic and environmental stochasticity. Haploid and monoecious or dioecious diploid populations are analyzed. Recent results from stochastic demography are employed to derive a two-dimensional diffusion approximation for the joint dynamics of the total population size, N, and the frequency of a selectively neutral allele, p. The infinitesimal variance for p, multiplied by the generation time, yields an expression for the effective population size per generation. This depends on the current value of N, the generation time, demographic stochasticity, and genetic stochasticity due to Mendelian segregation, but is independent of environmental stochasticity. A formula for the effective population size over longer time intervals incorporates deterministic growth and environmental stochasticity to account for changes in N. PMID- 15834151 TI - Multiple endonucleases function to repair covalent topoisomerase I complexes in Saccharomyces cerevisiae. AB - Topoisomerase I plays a vital role in relieving tension on DNA strands generated during replication. However if trapped by camptothecin or other DNA damage, topoisomerase protein complexes may stall replication forks producing DNA double strand breaks (DSBs). Previous work has demonstrated that two structure-specific nucleases, Rad1 and Mus81, protect cells from camptothecin toxicity. In this study, we used a yeast deletion pool to identify genes that are important for growth in the presence of camptothecin. In addition to genes involved in DSB repair and recombination, we identified four genes with known or implicated nuclease activity, SLX1, SLX4, SAE2, and RAD27, that were also important for protection against camptothecin. Genetic analysis revealed that the flap endonucleases Slx4 and Sae2 represent new pathways parallel to Tdp1, Rad1, and Mus81 that protect cells from camptothecin toxicity. We show further that the function of Sae2 is likely due to its interaction with the endonuclease Mre11 and that the latter acts on an independent branch to repair camptothecin-induced damage. These results suggest that Mre11 (with Sae2) and Slx4 represent two new structure-specific endonucleases that protect cells from trapped topoisomerase by removing topoisomerase-DNA adducts. PMID- 15834153 TI - Length and sequence heterozygosity differentially affect HRAS1 minisatellite stability during meiosis in yeast. AB - Minisatellites, one of the major classes of repetitive DNA sequences in eukaryotic genomes, are stable in somatic cells but destabilize during meiosis. We previously established a yeast model system by inserting the human Ha ras/HRAS1 minisatellite into the HIS4 promoter and demonstrated that our system recapitulates all of the phenotypes associated with the human minisatellite. Here we demonstrate that meiotic minisatellite tract-length changes are half as frequent in diploid cells harboring heterozygous HRAS1 minisatellite tracts in which the two tracts differ by only two bases when compared to a strain with homozygous minisatellite tracts. Further, this decrease in alteration frequency is entirely dependent on DNA mismatch repair. In contrast, in a diploid strain containing heterozygous minisatellite tract alleles differing in length by three complete repeats, length alterations are observed at twice the frequency seen in a strain with homozygous tracts. Alterations consist of previously undetectable gene conversion events, plus nonparental length alteration events seen previously in strains with homozygous tracts. A strain containing tracts with both base and length heterozygosity exhibits the same level of alteration as a strain containing only length heterozygosity, indicating that base heterozygosity dependent tract stabilization does not affect tract-length alterations occurring by gene conversion. PMID- 15834154 TI - Exploring the evolution of Wolbachia compatibility types: a simulation approach. AB - Wolbachia-induced cytoplasmic incompatibility (CI) is observed when males bearing the bacterium mate with uninfected females or with females bearing a different Wolbachia variant; in such crosses, paternal chromosomes are lost at the first embryonic mitosis, most often resulting in developmental arrest. The molecular basis of CI is currently unknown, but it is useful to distinguish conceptually the male and female sides of this phenomenon: in males, Wolbachia must do something, before it is shed from maturing sperm, that will disrupt paternal chromosomes functionality [this is usually termed "the modification (mod) function"]; in females, Wolbachia must somehow restore embryonic viability, through what is usually called "the rescue (resc) function." The occurrence of CI in crosses between males and females bearing different Wolbachia variants demonstrates that the mod and resc functions interact in a specific manner: different mod resc pairs make different compatibility types. We are interested in the evolutionary process allowing the diversification of compatibility types. In an earlier model, based on the main assumption that the mod and resc functions can mutate independently, we have shown that compatibility types can evolve through a two-step process, the first involving drift on mod variations and the second involving selection on resc variations. This previous study has highlighted the need for simulation-based models that would include the effects of nondeterministic evolutionary forces. This study is based on a simulation program fulfilling this condition, allowing us to follow the evolution of compatibility types under mutation, drift, and selection. Most importantly, simulations suggest that in the frame of our model, the evolution of compatibility types is likely to be a gradual process, with new compatibility types remaining partially compatible with ancestral ones. PMID- 15834155 TI - Kinetics of Src homology 3 domain association with the proline-rich domain of dynamins: specificity, occlusion, and the effects of phosphorylation. AB - Dynamin function is mediated in part through association of its proline-rich domain (PRD) with the Src homology 3 (SH3) domains of several putative binding proteins. To assess the specificity and kinetics of this process, we undertook surface plasmon resonance studies of the interaction between isolated PRDs of dynamin-1 and -2 and several purified SH3 domains. Glutathione S-transferase linked SH3 domains bound with high affinity (K(D) approximately 10 nm to 1 microm) to both dynamin-1 and -2. The simplest interaction appeared to take place with the amphiphysin-SH3 domain; this bound to a single high affinity site (K(D) approximately 10 nm) in the C terminus of dynamin-1 PRD, as predicted by previous studies. Binding to the dynamin-2 PRD was also monophasic but with a slightly lower affinity (K(D) approximately 25 nm). Endophilin-SH3 binding to both dynamin 1 and -2 PRDs was biphasic, with one high affinity site (K(D) approximately 14 nm) in the N terminus of the PRD and another lower affinity site (K(D) approximately 60 nm) in the C terminus of dynamin-1. The N-terminal site in dynamin-2 PRD had a 10-fold lower affinity for endophilin-SH3. Preloading of dynamin-1 PRD with the amphiphysin-SH3 domain partially occluded binding of the endophilin-SH3 domain, indicating overlap between the binding sites in the C terminus, but endophilin was still able to interact with the high affinity N terminal site. This shows that more than one SH3 domain can simultaneously bind to the PRD and suggests that competition probably occurs in vivo between different SH3-containing proteins for the limited number of PXXP motifs. Endophilin-SH3 binding to the high affinity site was disrupted when dynamin-1 PRD was phosphorylated with Cdk5, indicating that this site overlaps the phosphorylation sites, but amphiphysin-SH3 binding was unaffected. Other SH3 domains showed similarly complex binding characteristics, and substantial differences were noted between the PRDs from dynamin-1 and -2. For example, SH3 domains from c-Src, Grb2, and intersectin bound only to the C-terminal half of dynamin-2 PRD but to both the N- and C-terminal portions of dynamin-1 PRD. Thus, differential binding of SH3 domain-containing proteins to dynamin-1 and -2 may contribute to the distinct functions performed by these isoforms. PMID- 15834150 TI - Genetic linkage maps of the red flour beetle, Tribolium castaneum, based on bacterial artificial chromosomes and expressed sequence tags. AB - A genetic linkage map was constructed in a backcross family of the red flour beetle, Tribolium castaneum, based largely on sequences from bacterial artificial chromosome (BAC) ends and untranslated regions from random cDNA's. In most cases, dimorphisms were detected using heteroduplex or single-strand conformational polymorphism analysis after specific PCR amplification. The map incorporates a total of 424 markers, including 190 BACs and 165 cDNA's, as well as 69 genes, transposon insertion sites, sequence-tagged sites, microsatellites, and amplified fragment-length polymorphisms. Mapped loci are distributed along 571 cM, spanning all 10 linkage groups at an average marker separation of 1.3 cM. This genetic map provides a framework for positional cloning and a scaffold for integration of the emerging physical map and genome sequence assembly. The map and corresponding sequences can be accessed through BeetleBase (http://www.bioinformatics.ksu.edu/BeetleBase/). PMID- 15834156 TI - The adapter protein CrkII regulates neuronal Wiskott-Aldrich syndrome protein, actin polymerization, and tension development during contractile stimulation of smooth muscle. AB - Actin polymerization has been shown to occur in tracheal smooth muscle tissues and cells in response to contractile stimulation, and there is evidence that the polymerization of actin is required for contraction. In tracheal smooth muscle, agonist-induced actin polymerization is mediated by activation of neuronal Wiskott-Aldrich syndrome protein (N-WASp) and the Arp (actin-related protein) 2/3 complex, and activation of the small GTPase Cdc42 regulates the activation of N WASp. In the present study, the role of the adapter protein CrkII in the regulation of N-WASp and Cdc42 activation, actin polymerization, and tension development in smooth muscle tissues was evaluated. Stimulation of tracheal smooth muscle tissues with acetylcholine increased the association of CrkII with N-WASp. Plasmids encoding wild type CrkII or a CrkII mutant lacking the SH3 effector-binding ability, CrkII SH3N, were introduced into tracheal smooth muscle tissues, and the tissues were incubated for 2 days to allow for protein expression. Expression of the CrkII SH3N mutant in smooth muscle tissues inhibited the association of CrkII with N-WASp and the activation of Cdc42. The CrkII SH3N mutant also inhibited the increase in the association of N-WASp with Arp2, a major component of the Arp2/3 complex, in response to contractile stimulation, indicating inhibition of N-WASp activation. Expression of the CrkII SH3N mutant also inhibited tension generation and actin polymerization in response to contractile stimulation; however, it did not inhibit myosin light chain phosphorylation. These results suggest that CrkII plays a critical role in the regulation of N-WASp activation, perhaps by regulating the activation of Cdc42, and that it thereby regulates actin polymerization and active tension generation in tracheal smooth muscle. These studies suggest a novel signaling pathway for the regulation of N-WASp activation and active contraction in smooth muscle tissues. PMID- 15834157 TI - Molecular analysis of a store-operated and 2-acetyl-sn-glycerol-sensitive non selective cation channel. Heteromeric assembly of TRPC1-TRPC3. AB - We have reported that internal Ca2+ store depletion in HSY cells stimulates a nonselective cation current which is distinct from I(CRAC) in RBL cells and TRPC1 dependent I(SOC) in HSG cells (Liu, X., Groschner, K., and Ambudkar, I. S. (2004) J. Membr. Biol. 200, 93-104). Here we have analyzed the molecular composition of this channel. Both thapsigargin (Tg) and 2-acetyl-sn-glycerol (OAG) stimulated similar non-selective cation currents and Ca2+ entry in HSY cells. The effects of Tg and OAG were not additive. HSY cells endogenously expressed TRPC1, TRPC3, and TRPC4 but not TRPC5 or TRPC6. Immunoprecipitation of TRPC1 pulled down TRPC3 but not TRPC4. Conversely, TRPC1 co-immunoprecipitated with TRPC3. Expression of antisense TRPC1 decreased (i) Tg- and OAG-stimulated currents and Ca2+ entry and (ii) the level of endogenous TRPC1 but not TRPC4. Antisense TRPC3 similarly reduced Ca2+ entry and endogenous TRPC3. Yeast two-hybrid analysis revealed an interaction between NTRPC1 and NTRPC3 (CTRPC1-CTRPC3, CTRPC3-CTRPC1, or CTRPC1 NTRPC3 did not interact), which was confirmed by glutathione S-transferase (GST) pull-down assays (GST-NTRPC3 pulled down TRPC1 and vice versa). Expression of NTRPC1 or NTRPC3 induced similar dominant suppression of Tg- and OAG-stimulated Ca2+ entry. NTRPC3 did not alter surface expression of TRPC1 or TRPC3 but disrupted TRPC1-TRPC3 association. In aggregate, our data demonstrate that TRPC1 and TRPC3 co-assemble, via N-terminal interactions, to form a heteromeric store operated non-selective cation channel in HSY cells. Thus selective association between TRPCs generate distinct store-operated channels. Diversity of store operated channels might be related to the physiology of the different cell types. PMID- 15834158 TI - Bioconversion of isoeugenol into vanillin by crude enzyme extracted from soybean. AB - Crude enzyme extracted from soybean was used to convert isoeugenol into vanillin. The effects of several factors on the bioconversion were studied. Conversion was affected by the amount of substrate and was also improved by the addition of absorbents, among which powdered activated carbon was the best. The effect of H2O2 concentration on the conversion was also studied. The optimum concentration of H2O2 was 1% (v/v). With 10 g/L of powdered activated carbon and 0.1% H2O2 added, vanillin reached a maximum concentration of 2.46 g/L after 36 h, corresponding to a molar yield of 13.3%. PMID- 15834159 TI - Antioxidant responses to oleic acid in two-liquid-phase suspension cultures of Taxus cuspidata. AB - Two-liquid-phase plant cell cultures employ the use of a partitioning system to redirect extracellular product into a second phase. After the addition of organic solvent, in order to understand the defense system of Taxus cuspidata cells to organic solvent in two-liquid-phase suspension cultures, we investigated cells' antioxidant metabolism. The results showed that T. cuspidata cells responded to oleic acid with oxidative bursts in both intracellular H2O2 and extracellular O2 * production. Inhibition studies with diphenylene iodonium suggested that the key enzyme responsible for oxidative bursts was primarily NADPH oxidase. Investigation of the relationship between reactive oxygen species (ROS) and defense responses induced by oleic acid indicated that 4% (v/v) oleic acid increased the levels of antioxidant enzymes of superoxide dismutase, ascorbate peroxidase, and catalase and the antioxidant capacity of reduced ascorbate and glutathione. However, when oleic acid content reached a critical value (6% [v/v]), no further increase in antioxidant enzymes and antioxidant capacity was observed, indicating that the defense responses played a role in a certain range of oleic acid content, beyond which the overall ROS scavenging machinery was not induced and the peroxidation of membrane lipids emerged. PMID- 15834160 TI - Hydrolysis characteristics of tissue fractions resulting from mechanical separation of corn stover. AB - Corn stover has potential as a resource for both fiber and chemical needs if separation strategies can be developed to deal with its heterogeneity. Relative hydrolysis characteristics were assessed for pith (sclerenchyma and parenchyma) and fiber (collenchyma) tissue fractions derived from mechanical separation of corn stover to determine whether classification by tissue type resulted in fractions with different hydrolysis response. The physical characteristics of the tissue fractions were analyzed. The hydrolysis behavior of the fractions was evaluated under both acidic and basic conditions. The results from the hydrolysis experiments are compared with previously reported compositional analysis for the tissue fractions. PMID- 15834161 TI - Application of recombinant phage display antibody system in study of Codakia orbicularis gill proteins. AB - We used the recombinant phage display antibody system (RPAS) to obtain chimeric single-chain fragment variable (ScFv) antibodies to gill proteins of the white clam Codakia orbicularis (Linne, 1758). After three rounds of selection on immunotubes loaded with total gill protein extract, recombinant phages exhibiting antibodies to gill proteins were isolated and tested by enzyme-linked immunosorbent assay (ELISA). Clones exhibiting a high affinity for the mollusk proteins were selected for production of soluble ScFv antibodies, which were purified for subsequent analysis. ScFv antibodies exhibited a reaction specific for a protein whose molecular mass was about 15,000 Daltons and that was detected by the antigen capture technique followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting. PMID- 15834162 TI - An integrated process for separation of major and minor proteins from goat serum. AB - Analysis of minor proteins in animal sera is of considerable clinical significance. To be able to detect these proteins, depletion of major proteins (albumin and immunoglobulin G [IgG]) is necessary. Many of these proteins are also required in pure form for a variety of biochemical applications. The present work uses goat serum as the system and describes the separation and purification of both major and several minor proteins. This was carried out by judicious adaptation and combination of separation technologies such as immobilized metal ion affinity chromatography (on a somewhat novel matrix), dye affinity chromatography, and lectin affinity chromatography. Albumin, IgG, alpha2 macroglobulin, alpha1-proteinase inhibitor, and transferrin were obtained from the serum. The purified preparations were found to be homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis. PMID- 15834163 TI - Preparation of mono- and diacylglycerols by enzymatic esterification of glycerol with conjugated linoleic acid in hexane. AB - Esterification of glycerol with conjugated linoleic acid (CLA) was carried out in hexane. Lipase from Rhizomucor miehei provided a high degree of esterification (80%) in 8 h at 50 degrees C when used at 15% (w/w) in a system containing a 1:2 molar ratio of glycerol to free fatty acids. Esterification levels >80% were obtained in 8 h at 40 degrees C with 15% (w/w) lipase from Candida antarctica at the same molar ratio of reactants. The extent of esterification of CLA was >90% after 4 h of reaction at 50 degrees C with a 5% (w/w) loading of either R. miehei or C. antarctica lipase, together with a 1:1 molar ratio of substrates. Both enzymes incorporated the original CLA as acylglycerol residues in primarily 1,3 diacylglycerol and 1-monoacylglycerol. The CLA-rich acylglycerols can be employed as emulsifiers or as substitutes for natural fats and oils. PMID- 15834164 TI - Atopy and asthma prevention: new paradigms? PMID- 15834165 TI - Primary prevention of allergy and asthma is possible. AB - Allergic diseases, such as asthma, are the result of complex gene-environment interactions. The focus for prevention of allergic disease has been on environmental control. Environmental exposures begin during the intrauterine period, during which a T-helper-2 immune phenotype is promoted. Food allergy frequently occurs during the first year of life, and avoidance of exposure must begin in early pregnancy and must be complete to be effective. Partial avoidance strategies have not been successful. Current data do not allow us to specifically recommend breastfeeding for the prevention of allergy and allergic diseases, but for other important reasons, breastfeeding (particularly exclusive breastfeeding) should be encouraged for at least the first 4 mo of life. Sensitization to allergens is one of the strongest determinants for subsequent development of asthma. There is a strong relationship between exposure to house dust mites, allergen sensitization, and asthma. However, exposure to pets and animals in a farming environment early in life may actually be protective for the development of allergy and asthma. Specific recommendations relating to these exposures requires additional research, but genetics clearly plays an important role in that process. To date, only a multifaceted intervention program has been successful as a primary prevention strategy for the development of asthma in young children. The specific components of the multifaceted intervention and the duration of protection have not yet been defined. It is increasingly clear that gene-directed environmental manipulation undertaken in a multifaceted manner during a "window of opportunity" is critical in the primary prevention of allergy and allergic diseases like asthma. PMID- 15834166 TI - Allergen avoidance as primary prevention: con. AB - There has been a dramatic increase in the prevalence of atopic diseases, such as atopic dermatitis, food allergy, allergic rhinitis, and allergic asthma. Because environmental factors such as early allergen exposure to nutritional and inhalant allergens have been suspected as potential environmental factors favoring the development of allergic diseases and asthma, primary prevention studies have begun to investigate the effect on sensitization and wheezing. Designs of the studies are different and, therefore, are not completely comparable. Although no clear primary prevention study is available for pets, mite allergen avoidance during infancy can achieve a reduction of specific sensitization and, in some studies, can also produce a slight effect on respiratory symptoms. PMID- 15834167 TI - High allergen exposure as a risk factor for asthma and allergic disease. AB - The association between pet ownership in childhood and subsequent asthma and sensitization is very controversial. Intriguing, but contradictory, reports have caused considerable uncertainty in parents who wish to avoid asthma and allergic disease in their children. This article argues that high allergen exposure is a risk factor for asthma and allergic disease. It describes dispersal of pet allergens in society and critically assesses epidemiological studies regarding how early exposure to pet allergens affects subsequent immunoglobulin E sensitization and allergic diseases. Additionally, this article evaluates the effects of allergen exposure in already sensitized subjects with asthma. Cat and dog allergens are ubiquitous in society and may induce sensitization and allergic symptoms in predisposed individuals, regardless of pet ownership. This, in combination with selection mechanisms for pet ownership in families with a history of allergic diseases, makes it difficult to study associations between early exposure to pets and subsequent allergic disease. Nevertheless, exposure to pet allergens worsens asthma in already sensitized children. Thus, it is clear that clinicians should advise sensitized asthmatics that avoidance of exposure to indoor allergens is an important element in the treatment of allergic disease. PMID- 15834170 TI - [Use of less invasive techniques in surgical treatment of dumping syndrome]. AB - Aiming to minimize the surgical trauma and interventional risks of treatment, as well as to reach the most precise visualization and safe separation of the n. vagus, in order to increase the efficacy of truncal vagotomy during correction of severe post-gastric-resection dumping syndrome we have developed the new method of thoracoscopic supradiafragmal vagotomy. The suggested method of bilateral thoracoscopic supradiafragmal truncal vagotomy allows to decrease the surgical intervention trauma, and the risk of post operative complications, as well as to better the immediate and remote results of treatment of the dumping syndrome. PMID- 15834168 TI - High-dose allergen exposure leads to tolerance. AB - Reports of decreased sensitization to cat allergen (Fel d 1) among individuals living with a cat or subjects exposed to high-dose cat allergen may be explained by the development of a form of high-dose tolerance resulting from natural exposure to an inhalant allergen. Although the epidemiological data regarding the relationship between exposure and sensitization to Fel d 1 are conflicting, the ability for high-dose Fel d 1 to induce a characteristic nonallergic immune response with a distinctive serum antibody profile has been established. Definition of this modified T-helper (Th)2 response to cat allergen, coupled with the renewed interest in regulatory T cells within the immunology field, has provided an avenue for exploring the mechanism by which IgE antibody-mediated responses are controlled. There is mounting evidence to suggest that the modified Th2 response is a variation of the allergic response and that the modified Th2 allergic axis is influenced by allergen dose and genetics. This article discusses putative immune mechanisms of tolerance within the context of an allergen specific system. The relevance of high-dose allergen exposure and alternate factors such as endotoxin to the development of tolerance is considered. Fel d 1 exhibits unique molecular and immunological characteristics that may contribute to its tolerogenic properties. Major T-cell epitopes of Fel d 1 that preferentially induce regulatory factors have been defined. Furthermore, high titer IgE antibody responses associated with atopic dermatitis are characterized by a defect in the T-cell repertoire that is specific to these epitopes. Identification of Fel d 1 epitopes that induce interleukin-10 may provide new targets for treatment. PMID- 15834169 TI - Possible reasons for lack of effect of allergen avoidance in atopy-prone infants and sensitive asthmatic patients. AB - The basic paradigm that allergen exposure produces atopic sensitization, and that continued exposure leads to clinical asthma throughout the development of airway inflammation and bronchial hyperreactivity has been challenged. However, because it was observed that epidemiological evidence suggests that around 40% of asthma cases are attributable to atopy (even using restrictive criteria), the obvious corollary is that if allergen avoidance begins before the onset of sensitization (primary prevention), then it should be associated with a reduced number of new cases of the disease. However, there are conflicting results regarding the effect of allergen avoidance on primary prevention of atopic sensitization and asthma onset. Instead, more uniform and positive results are available from secondary prevention studies. Secondary prevention obviously is an attractive opportunity for pediatricians who may recognize the patients who might benefit from these interventions simply by screening for food allergy in young children with atopic dermatitis. The conflicting results of tertiary prevention are most frequently observed in adult patients and sometimes result from incomplete avoidance of allergens responsible for the sensitization. PMID- 15834171 TI - [Effect of the metabolic syndrome on the electrophysiological parameters of the heart]. AB - The goal of this research was to study influence of the MS on the electrophysiological parameters of the heart conduction system. The research involved 32 patients suffering from heart arrhythmias (18 females and 14 males, average age 52,8-/+12,9). The patients were categorized into 2 groups. Group I included 15 patients (9 females and 6 males, average age 47,5-/+8,9) suffering from heart arrhythmias and with signs of MS. Group I was subdivided into 2 subgroups: I subgroup--10 patients without diabetes, and II subgroup 5 patients with type II diabetes. Group II comprised of 17 patients (8 females and 9 males, average age 57,4-/+11,9) with arrhythmias without any signs of MS. The refractoriness dispersion between the right atrium (RA) and the left atrium (LA) caused by the MS, directly provokes development of atrial fibrillation and type II (atypical) atrial flutter. In cases of metabolic syndrome and "slow-fast" type of PRAVNT caused by the metabolic failures, the prolongation of the antegrade effective refractory period (ERP) of the slow AV nodal pathways and the retrograde ERP of the fast AV nodal pathways provoked prolongation of the tachycardia cycle and, consequently, reduction of the heart rate during paroxysms of tachycardia. Metabolic failures produce direct effect upon the sinus node function, causing suppression of its function as the SNSS develops, which requires implantation of pacemaker. The research results for metabolic syndrome and AV blockage cases demonstrated that the latter was always of a distal type, which was caused by a direct effect of metabolic failures upon the electrophysiology of the atrioventricular junction. PMID- 15834172 TI - [Detection of mycobacteria tuberculosis in patients with urogenital tuberculosis by PCR method]. AB - The study was carried out in hospital patients as well as in outpatients at the National Centre of Tuberculosis and Lung Diseases of Georgia (2002-2004). The group consisting of 32 patients with tuberculosis of urogenital system has been studied (newly detected forms). Except clinical laboratory, culture and X-ray contrast methods, two additional methods were used in testing of this group of patients. The examination of their urine, at the same time, was carried out by the Polymerase Chain Reaction method in order to detect Kochi bacillus and by three-time bacterioscopy of urine for acid resistant bacteria. Mycobacterium tuberculosis in urine has been detected in 26 (81,25%) patients by PCR method, and by urine bacterioscopy--acid fast bacilli (AFB+) in 18 (56,25%) patients. The histo-morphological investigation of specimens obtained by surgery confirmed the TB diagnosis in all patients. This study on patients suspected of Tuberculosis of genital-urinary system gives us an opportunity to update the diagnostic algorithm by including the modern molecular methods. This algorithm will help in timely detection of Tuberculosis, in selection of adequate therapy and in prevention of the further progression of the disease. PMID- 15834173 TI - [Role of follicular fluid analysis in assessment of the main criteria of folliculogenesis in IVF program]. AB - A comparative study was carried out to assess the main criteria of folliculogenesis in follicular fluid of aspirated follicles during the induction of super ovarian stimulation using recombinant follicle stimulating hormone (r FSH; Gonal-F), human menopausal gonadotropin (HMG, Pergonale) and agonist gonadotropin releasing hormone (a-GnRH; Diphereline 3,75 mg). 86 patients were included in the study: 37 were receiving r-FSH and 49 HMG, two ampoules per day during the first 5 days of stimulation. Ultrasound monitoring was used for every individual follicle on the first day of stimulation and starting from the 5th day, daily. Thus, the visualized follicles were identified and measured, photoregistered during the whole period of ultrasound guidance. During the transvaginal puncture of these follicles the aspirated follicular fluid volume, existence of oocytes, and its estradiol hormones were recorded. It was found that the follicles in both groups of patients having larger volume of follicular fluid (3 to 5 or >5mm) reached the preovulational size earlier (diameter 17 mm), and on the day of injection of human chorionic gonadotropin (HCG; 10000 IU) had a large size (20 to 21 mm). The positive statistically significant correlation has been observed between the volume of follicular fluid and existence of oocytes in aspirated follicular fluid, as well as with the intensity of their cleavage. The concentration of testosterone and estradiol in follicular fluid was lower in the patients receiving r-FSH (average 12.4 nmol/l, range 9.3-15.1 and 1897113 pmol/l; range 1609216-2185012) than in the patients receiving HCG (16.0 nmol/l; range 12.1-19.6 and 2233728 pmpl/l; range 2028660-2438718). The concentration of testosterone in non-pregnant patients receiving r-FSH was 13.7-/+1.0 nmol/l, receiving HMG - 16.0-/+0.9 nmol/l, versus 10.3-/+0.8 nmol/l and 14.8-/+0.7 nmol/l, in the patients receiving r-FSH and HMG, respectively, who became pregnant. Thus, the comparative study of main criteria of folliculogenesis in follicular fluid proved to be more preferable and sufficient using FSH in the ovarian stimulation protocol in IVF program. PMID- 15834174 TI - [Determination of cytomorphological criteria for differential diagnostics of variable stages of endocervical glandular cells atypia]. AB - The aim of the investigation was to determine the criteria of differential diagnostics of different stages atypia of endocervical cells in women of different age groups. For ECCIN3, out of 12 diagnostic criteria, the most important and statistically significant were the following five: a) high N/C index > 5,54; b) number of mitoses > 10,3; c) number of produced rosettes >3,9; d) rough redistribution of chromatin >4,64; e) hyperchromatosis >1,87. In case of ECCIN3 the majority of patients (82,75%) had infection of the cervix. In 13,79% of patients HPV infection has been documented. Hemorrhagic discharge was a very characteristic symptom (57,12%) of ECCN3. PMID- 15834175 TI - [Significance of ultrasound assessment of placental tissue in diagnostics of post term pregnancy]. AB - The purpose of this work was to determine the value of ultrasound investigation of the placental tissue in fetal assessment during post-term pregnancy. 80 pregnant women from 39 weeks gestation till delivery were examined in this long term study. The investigation was performed every 5-7 days until the labor. They were divided into three groups. The first group included 40 pregnant women with post term pregnancy, the second, 30 women with prolonged pregnancy. The control group consisted of 10 practically healthy pregnant women. Ultrasound parameters of fetal biometry such as the BPD, APTD, TTD and dynamics of placental thickness changes were analyzed. Our results suggest that the post term pregnancy can be differentiated from prolonged pregnancy by ultrasound assessment of placental tissue (monitoring of changes in placental thickness) and results of fetal biometry (BPD, APTD, TTD). PMID- 15834176 TI - [Sentinel nodes identification efficacy prognostic criteria in breast cancer]. AB - The sentinel node concept assumes that a malignancy that metastasizes by the lymphatic route will initially travel to one or a few lymph node(s). Although a tumor may disseminate to other nodes, it will remain in the sentinel node, and therefore the status of the entire basin is predicted by the sentinel node. The sentinel lymph node biopsy with total axillary dissection was performed on 99 patients. The identification rate was 69,7%, specificity 62,6%, and the false negative rate was 7,1%. The impact of some prognostic factors on efficacy of visualization was also investigated. It was shown that the age of patients, stage of disease, localization of primary tumor, and method of injection of methylene blue were important prognostic factors for effective identification of sentinel lymph nodes in breast cancer patients. PMID- 15834177 TI - [Treatment of sclerotic stenosis of trachea]. AB - An evident trend towards the increase of postintubational and posttracheostomic tracheal stenoses is observed in Georgia during the past 10-12 years. For example, during the period of 1998-2001 28 patients were admitted to our center with this diagnosis, while during 2001-2004 we treated 61 similar cases. According to several international publications overall frequency of postventilatory laryngotracheal stenoses is as high as 25%. At present time surgical intervention remains as a method of choice for treatment of tracheal stenoses including circular resection, step by step reconstructive plastic operations and different methods of tracheal recanalization. 89 patients 14-68 years of age were treated in our center during 1998-2004. 61 (68,5%) cases were presented with pathology within the cervical part of trachea and larynx. 28 (31,5%) cases were with intrathoracic tracheal stenosis. In 71 (79,8%) cases stenoses were induced by pulmonary ventilation by endotracheal tube and in 18 (20,2%) cases etiologic factor was posttracheostomic tracheal injury. Duration of intubations varied from 3 to 86 days. Mortality was 3 cases out of 89. In 2 cases mortality was due to anastomotic leakage, in 1 case--bleeding from brachiocephalic vessels. Based on our experience, an accurate follow up of diagnostic and treatment algorithms allows achievement of good results in 91% cases. PMID- 15834178 TI - [The immunomorphological characteristics of chronic recurrent ulcerative stomatitis]. AB - The aim of the present study was to evaluate the immunomorphological characteristics of chronic recurrent ulcerative stomatitis. The patients were divided into 3 groups: 16 patients with mild, 15 patients with moderate, and 9 with severe stomatitis. The cytological smears were stained by Papanicolaou method. We have evaluated indexes of maturation (MI), keratinization (KI), destruction (DI), and inflammation-destruction (IDI). The immunocytochemistry was used to evaluate the local immune reactions. Anti-CD20 (pan-B marker), Anti-CD3 (pan-T marker), CD4 (marker of T helper), CD8 (marker of T cytotoxic lymphocytes) monoclonal antibodies were used (LSAB, DAB). The comparative analysis of cytological indexes in acute and remission phases of chronic recurrent ulcerative stomatitis showed that DI and IDI decreased in remission phase compared with acute phase, but did not return to norm. It seems that inflammation persists in remission phase despite the absence of symptomatic ulcerative lesion. In mild chronic recurrent ulcerative stomatitis the dynamic quantitative changes of immunocompetent cells in acute and remission phases show that the increased number of CD4+ T and CD20+ B lymphocytes has been found in remission phase indicating a persistent immune reaction and presence of non-eliminated pathogenic factors. PMID- 15834179 TI - [Bar construction upon the implants--successful method of treatment of toothless jaws]. AB - Frontal parts of jaws are the most convenient for invasion of sufficient number of supporting implants that can solve the problem of fixation of dentures (prosthesis). Bar Construction upon the implants that have supporting attachments are one of the best fixation systems of removable dentures (prosthesis) on toothless jaws white damaging alveolar ridge. Elastic metal patrice is the same kind of fixation. But the priority of it is the simplicity and cost effectiveness. We have discussed one clinical history. We have made implantation to 78 years old patient, and then we made bar construction and elastic metal patrice for him. After 6 years it will be clear whether this type of construction is successful or not. PMID- 15834180 TI - Pleiotropic effects of cardilopin (secondary coronary prevention). AB - In order to evaluate some pleiotropic effects of cardilopin (Amlodipine, EGIS Pharmaceuticals) 33 ambulatory patients were examined. During the 2-month study period on the background of cardilopin treatment there were positive changes in some parameters of coronary atherosclerosis. Pleiotropic effects of cardilopin were particularly expressed in restoring of endothelial function and inhibition of platelet aggregation. There was found tendency to reduce the degree of hyperlipoperoxidemia related to oxidative stress. Obtained results of the present trial support the use of cardilopin in all coronary heart disease patients with or without myocardial revascularization and arterial hypertension. PMID- 15834181 TI - Hemopoetic organs damage syndrome in patients with chronic viral hepatitis. AB - Liver cirrhosis and hepatocellular carcinoma are the most frequent and severe complications of chronic hepatitis C. Results of recent investigations indicate that HCV persists and replicates not only in hepatic tissue, but in the other organs as well. The goal of our investigation was to determine hematological disorders during chronic HCV and HCV+HBV hepatitis, to find correlation between activity of chronic hepatitis, stage of cirrhosis, and degree of hematological disorders. We have studied patients with chronic C and B+C hepatitis (36 persons) and 77 HCV-infected persons on the stage of cirrhosis. The erythrocytes morphology was investigated using light microscope. The osmotic resistance of erythrocytes was investigated using the method of Ianovski (modification of Ribiera); the thrombocytes' ability to adhere and aggregate was studied by Rite's method. The absolute number of thrombocytes was counted by means of phase contrast microscope. Our investigation suggests that there is no absolute correlation between chronic hepatitis C activity and hematological disorders and often the degree of extrahepatocellular disorders is higher than the degree of liver damage. PMID- 15834182 TI - [Parameters of circadian heart rate variations, night sleep profile and structure during the Holter monitoring procedure in the groups of students with different adaptation potential]. AB - Circadian heart rate variations, night sleep profile and structure during ECG Holter monitoring process have been studied in practically healthy students with various adaptation potential to estimate the role of the dynamics of these parameters for evaluation of functional condition of cardiovascular system and whole organism. Statistically significant differences have been revealed for the functionally distinguished groups for the following parameters: P average/day, Circadian Index, sleep profile and sleep duration. When the mechanisms of adaptation are very strained, P average/day is high and circadian profile is strengthened, which indicates to the increase of sympathetic influence on the heart rhythm in persons who have primary high level vagotonia. With increasing of the strength of the adaptation mechanisms the sleep duration (sleep profile as well) is reducing, which may be one of the causes of asthenization. PMID- 15834183 TI - [Activity of lipid peroxidation processes in children with rheumatic fever]. AB - Pathogenic mechanism of acute and chronic inflammation is connected to the increased production of superoxide anion, hydrogen peroxide, and other free radicals. Clinical role of lipid peroxidation (LPO) processes was studied in 38 patients in the age from 3 to 15 years old with different variants of Rheumatic Fever (RF). We have investigated the relationship between malonidialdehide (MDA) and RF. We measured the levels of MDA in the plasma in patients with acute rheumatic fever (ARF) and chronic rheumatic heart disease (CRHD). Our study revealed that the levels of MDA in patients with ARF are significantly higher than in patients with CRHD. These levels were also significantly higher in patients with history of disease for up to 2 years, than in patients with history longer than 2 years. Thus, the measurement of MDA in the plasma could be used as a laboratory test for relation of active state of rheumatic fever. PMID- 15834184 TI - [Lymphatic system of the tongue and its role in glositis of odontogenic origin]. AB - In aged persons reduction of diameter of tongue lymphatic capillaries precedes thinning of the Kaarl net. In the process of tongue inflammation, lymphogenic way of inclusion in 6|6 and 8|8 teeth lower area should be stuck out with existence of alleged "Integration Centers". Lymphatic knots and lymphatic ducts are in prevailed placed in corresponding tissues of lower-chin and lower teeth. Lymphatic-muscular system and its anatomical links and age-related changeability raise the special interest during odontogenic infections with tongue inflammation. PMID- 15834186 TI - [Immunohistochemical investigation of steroid hormones (ER, PR) in breast cancer]. AB - Immunohistochemical investigations were performed in 22 cases of breast cancer. The objective of this study was to assess the presence and distribution of ER and PR in the cancer tissue. We found that expression of the ER and PR in the cancer tissue is variable, which depends on cancer histogenesis (lobular, ductal), histological form, differentiation grade, and stage of its development. Level of the expression of ER and PR was higher in the ductal form than in the lobular form of breast cancer. Expression of the ER was significantly higher than expression of PR in the case of the lobular form. There were no similar differences seen in cases of ductal form of breast cancer. PMID- 15834185 TI - [Plaferon LB in prevention of radiation-induced changes in lipid metabolism indices and erythrocyte deformability rate]. AB - Oxidative damage of biological membranes plays critical role in the process of radiation-induced damages in tissues and cells. It involves reactive oxygen species generated under ionizing radiation. Development of radiation-induced oxidative stress facilitates the intensification of lipid peroxidation and fatty acid reorganization, which, in its turn, results in decreased lipid matrix fluidity, changes in biophysical properties of membranes, increased rigidity of membrane lipid-protein surface. Radiation-induced damage of cellular and subcellular membrane structures results in cellular metabolism disorder, which facilitates farther dysfunction of tissues and changes in homeostasis of whole organism. Hence, the search for effective protection is of actual interest of contemporary radiology. Our study aimed to determine membrane protective properties of Plaferon LB under radiation. On the basis of obtained results we can conclude that intensification of free radical oxidation, accompanying radiation damage, facilitates the activation of lipolysis in a body and functional and structural disorders of erythrocytes revealed by lowered deformability of erythrocytes and accumulation of methaemoglobin. Both vitamin C and Plaferon LB were found ineffective for correction of lipid metabolism at early stage of post radiation damage; although facilitating a decrease in oxidation intensity in blood, they provided retention of erythrocyte deformability and thereby, partial retention of rheological properties of blood. It should be mentioned that in contrast with vitamin C, Plaferon LB, apart from its ability to restrict reactive oxygen formation, revealed ability to stabilize erythrocyte membranes and thereby to prevent hemolysis. PMID- 15834187 TI - [Association between the hemodynamic and rheological parameters in the micro blood vessels in vivo]. AB - Considering the experimental data obtained earlier, regarding the shift velocity in the micro blood vessels in vivo, and general theses of liquid mechanics and rheology of the non-Newtonian liquids, a generalized ratio between the hemodynamic and rheological parameters in the micro blood vessels in vivo are overviewed. These are: pressure difference--DeltaP, volume of blood flow--Q, linear velocity of blood flow, viscosity of Newtonian liquid--k, effective viscosity of the blood in a vessel in vivo eta(effect), and index of rheological behavior--n, which was determined earlier in our laboratory. These parameters are observed in the live blood vessels of microcirculatory bed, in all regimes of the blood flow characteristic for the in-vivo vessels. PMID- 15834188 TI - Histological and histochemical investigations of nasal septum cartilage in nasal deformations. AB - AIM of the study is to identify histological and histochemical peculiarities of nasal septum cartilage reconstruction in patient with different remoteness of trauma. MATERIALS AND METHODS: Patients of both sexes at the age of 16-39 years have been operated on. Depending on remoteness of the period from the trauma, witch caused the deformation, till the surgery, the materials was divided into 2 group: I group (36 patients) 1-5 years from the trauma to the surgery, II group (23 patient) 10-20 years from the trauma to the surgery. Bioptats of the nasal septum cartilage were fixed in Carnoy mixture. Paraffin sections were stained with hematoxilin and eozin, after Van Ghison on collagen fibrils, toluidine blue on acid glycoaminglicans (GAG) and Brashe reaction on RNA. STUDY RESULTS AND CONCLUSION: Thus, the histological and histochemical investigation of nasal septum cartilage, received in rhinoplasty, has revealed dystrophic changes in cartilage. Basing on the analysis of the material, we can conclude that the degree of expression of these changes depends on three factors: - localization of examined area of the nasal septum (bioptat); - the period of time the deformation exist; - the degree of septum curvature. The structure of cartilage bioptates in the patients with relatively short (1-5 years) duration of traumatic deformation differs from the normal hyaline cartilage by moderate dystrophic changes of some chondrocytes (20-25% of all the cells) in the central zone of the cartilage as well as by irregular distribution of acid GAG in cartilage matrix. Cartilage tissue dystrophic changes are still more pronounced in the patients with a longer period of nose deformation including nasal septum curvature. This fact indicates that these changes are connected with deformation duration rather than with etiology of the process. The analysis of material also enable to find the dependence of cartilage dystrophic changes on the degree of nasal septum deformation: at the 3rd degree of curvature, chondropathy is more pronounced than at the 1st-2nd degree. PMID- 15834189 TI - The President's Council on Bioethics 2002-2004: an overview. AB - The President's Council on Bioethics, headed by Leon Kass, was created by President George W. Bush to advise the President on issues of ethical import raised by advances in biomedical science. Between 2002 and 2004, members of the Council from diverse disciplines addressed topics such as human cloning, stem cell research, assisted reproduction, and medical interventions intended to enhance human capability or appearance. This article provides background on the Council and reviews its published reports. It also considers key definitions and distinctions, specific recommendations of the Council, and positions articulated by members who contributed to the development of its reports. PMID- 15834190 TI - Thoughts of a former Council member. PMID- 15834191 TI - Stem cell research: the bigger picture. PMID- 15834192 TI - Human biomedicine and the problem of governance. PMID- 15834193 TI - The moral status of the human embryo. PMID- 15834194 TI - Altered nuclear transfer as a morally acceptable means for the procurement of human embryonic stem cells. PMID- 15834195 TI - The President's Council on Bioethics: my take on some of its deliberations. PMID- 15834196 TI - The ethical implications of human cloning. PMID- 15834197 TI - Pain and the placebo: what we have learned. AB - Despite the recent blossoming of rigorous research into placebo mechanisms and the long-standing use of placebos in clinical trials, there remains widespread and profound misunderstanding of the placebo response among both practicing physicians and clinical researchers. This review identifies and clarifies areas of current confusion about the placebo response (including whether it exists at all), describes its phenomenology, and outlines recent advances in our knowledge of its underlying psychological and neural mechanisms. The focus of the review is the placebo analgesic response rather than placebo responses in general, because much of the best established clinical and experimental work to date has been done on this type of placebo response. In addition, this subfield of placebo research offers a specific neural circuit hypothesis capable of being integrated with equally rigorous experimental work on the psychological (including social psychological) and clinical levels. In this sense, placebo analgesia research bears all the marks of a genuine multilevel interdisciplinary research paradigm in the making, one that could serve as a model for research into other kinds of placebo responses, as well as into other kinds of mind-body responses. PMID- 15834198 TI - The remarkable vision of Robert Hooke (1635-1703): first observer of the microbial world. AB - Robert Hooke played important roles in the early development of the Royal Society of London. As Curator of Experiments of the Society, he became a pioneering microscopist, prolific inventor, astronomer, geologist, architect, and an effective surveyor of the City of London following the Great Fire of 1666. Hooke's Micrographia (1665) revealed the microscopic structures of numerous biological and inorganic objects and became an important source of information for later studies. Aside from the body of detailed observations reported and depicted in Micrographia, the Preface is in itself an extraordinary document that exhibits Hooke's fertile mind, philosophical insights, and rare ability to look into the future. PMID- 15834199 TI - William James, faith, and the placebo effect. AB - Long considered a fact of medicine and of clinical investigation, the placebo effect has recently been challenged. The thought of the great American psychologist and philosopher William James, particularly his understanding of the practical value of faith, helps to illuminate the nature of the placebo effect and the implications of this puzzling phenomenon for understanding healing and the practice of medicine. PMID- 15834200 TI - When medical cure is not an unmitigated good. AB - The impact of chronic illness on patients has been discussed extensively, but little attention has been paid to the reverse situation: the need for patients who have been chronically ill since childhood to adjust to health and normality in adulthood. Drawing upon the narratives of two adults with childhood onset diseases, this paper provides a new perspective about the transition from chronic illness to health after a successful transplantation. We analyze from the patients' perspective the unusual disruptive impact of overcoming illness in such situations. The process of becoming cured affects their identity, leaving them unable to restructure their self within health and normality, in the same way as others are unable to adjust to chronic illness. Health care providers and families must pay more attention to the losses and life changes these patients undergo after being cured, and to their specific needs in the post transplantation situation. We hope to offer insight into ways to prevent and address the difficulties and suffering in such situations, which may become more frequent as more therapeutic options become available for adults with childhood onset diseases. PMID- 15834201 TI - Pharmacological modulations in human cognitive processes: an fMRI study. PMID- 15834202 TI - Free radical development in phacoemulsification cataract surgery. AB - Phacoemulsification and aspiration (PEA) has become the most popular cataract surgery, due to the establishment of safe surgical techniques and development of associated instruments. However, corneal endothelial damage still represents a serious complication, as excessive damage can lead to irreversible bullous keratopathy. In addition to causes such as mechanical or heat injuries, free radical formation due to ultrasound has been posited as another cause of corneal endothelium damage in PEA. Ultrasound in aqueous solution induces cavitation, directly causing water molecule disintegration and resulting in the formation of hydroxylradicals, the most potent of the reactive oxygen species. Considering the oxidative insult to endothelial cells caused by free radicals, their presence in the anterior chamber may represent one of the most harmful factors during these procedures. Indeed, some researchers have recently started to evaluate PEA from the perspective of oxidative stress. Conversely, the major ingredient in ophthalmic viscosurgical devices (OVDs), which are indispensable for maintaining the anterior chamber in PEA surgery, is sodium hyaluronate, a known free radical scavenger. OVDs can thus be expected to provide some anti-free radical effect during PEA procedures. In addition, since commercially available OVDs display different properties regarding retention in the anterior chamber during PEA, the anti-free radical effect of OVDs is likely to depend on behavior during irrigation and aspiration. The present study followed standard PEA procedures in an eye model and measured hydroxylradicals in the anterior chamber using electron spin resonance. The kinetics of free radical intensity and effects of several OVDs during clinical PEA were also demonstrated. These studies may be of significance in re-evaluating OVDs as a chemical protectant for corneal endothelium, since the OVD has thus far only been regarded as a physical barrier. In addition, many reports about corneal endothelium damage during PEA have been published, but objective evaluation of various damaging factors has been difficult. The present assay of free radicals in a simulation of clinical PEA offers the first method to quantitatively assess stress on the corneal endothelium. PMID- 15834203 TI - Epithelial-connective tissue cross-talk is essential for regeneration of intestinal epithelium. AB - Epithelial cells of the gastrointestine undergo a rapid cell-renewal and originate from stem cells throughout the life of the organisms. Previous studies have provided a solid body of evidence to show that the epithelial cell-renewal is under the strict control of cell-cell and cell-extracellular matrix (ECM) interactions between the epithelium and the connective tissue. Especially, the microenvironment around the stem cells called "niche" is thought to play important roles in this control, and its disruption leads to diseases or disorders such as cancer in the human gastrointestine. Although understanding how the niche affects the stem cells is clinically important, its mechanisms still remain mostly unknown at the molecular level, possibly due to difficulties in the identification of the stem cells in the gastrointestine. Recent progress in cell and molecular biology is gradually beginning to shed light on some of the key signaling pathways in the cell-renewal of the intestinal epithelium, such as Wnt/T-cell factor (TCF)/beta-catenin, Notch, Sonic hedgehog (Shh)/bone morphogenetic protein (BMP) signaling pathways, which are also involved in embryonic organogenesis and/or adult carcinogenesis. At present, only fragmentary information is available on their precise functions in the intestine. Nevertheless, there is a growing body of evidence that such signaling pathways have conservative functions in the intestine throughout terrestrial vertebrates, suggesting the usefulness of experimental animals to clarify molecular mechanisms regulating epithelial cell-renewal. In this article, I review some recent findings in this field, with particular focus on our studies using the Xenopus laevis intestine, where the stem cells form the mammalian-type intestinal epithelium under the control of connective tissue during metamorphosis. This Xenopus experimental system will certainly serve as a useful model for the study of the intestinal niche, whose clarification is urgently needed in regenerative medicine. PMID- 15834204 TI - Does carboxy-hemoglobin serve as a stress-induced inflammatory marker reflecting surgical insults? AB - Endogenous carbon monoxide (CO) production has been recently observed to be an index of the inflammatory response, reflecting various insults in critically ill patients. Major surgery is supposed to modulate the production of CO by transcriptional regulation of heme oxygenase (HO). CO is easy to measure as carboxyhemoglobin (CO-Hb) by spectrophotometry; however, whether CO-Hb can be used as an index reflecting surgical insults is unknown. We investigated changes in CO generation during coronary artery bypass graft by measuring CO-Hb concentrations and the expression of HO in circulating blood as well as the expressions of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL 1 beta). The expression ratios of heme oxygenase-1 (HO-1), TNF-alpha, and IL-1 beta significantly increased after surgery, and these values correlated significantly with one another. CO-Hb concentrations significantly increased after surgery; however, many of those values during artificial ventilation with high inspired oxygen fraction were within normal limits. Furthermore, changes in CO-Hb concentrations were small when preoperative values were high. On the whole, CO-Hb concentrations significantly but weakly correlated with the expression ratios of the inflammatory mediators. However, they did not correlate in the patients who showed higher preoperative CO-Hb concentrations. These data indicate that CO-Hb concentrations can, in general, reflect the inflammatory response induced by surgical insult; however, CO-Hb measurement may not be a useful form of clinical monitoring because of the limited degree of changes, the variation of baseline values, and the necessity for the management under fixed conditions. PMID- 15834205 TI - Calcifications in mucinous and serous cystic ovarian tumors. AB - Mucinous cystic ovarian tumors sometimes contain calcifications, but the frequency and significance of such calcifications in diagnostic radiology are not well understood. We therefore retrospectively investigated the radiological and histopathological evidence of calcifications in 44 cases of ovarian mucinous cystic tumors (22 benign, 13 borderline, and 9 malignant) and 21 cases of ovarian serous cystic tumors (6 benign and 15 malignant) in which a non-contrast CT scan was performed. The shape and distribution of the calcifications in the mass lesion were assessed both radiologically and histopathologically. Calcifications were noted in 34.1% of mucinous cystic tumors on CT scans and 56.8% in histopathological studies, and they were found in two locations, intramural and intra-cystic, according to the histopathological findings. Intramural calcifications were frequent in benign tumors, and intra-cystic calcifications were frequent in proliferating tumors. Calcifications (psammoma bodies) were noted in 4.7% of serous cystic tumors on CT scans and 14.3% in histopathological studies. CT was not sufficiently sensitive in the detection of intra-cystic calcification in mucinous tumors and psammoma bodies in serous tumors. However, the presence of intramural calcifications may be a good indicator of mucinous tumors. Understanding the frequency and morphology of the calcifications in these neoplasms is one of the keys to making a correct diagnosis. PMID- 15834206 TI - Thoracoscopic surgery in children. AB - PURPOSE: A minimally invasive thoracoscopic surgery offers several options in diagnosis and surgical treatment in the field of pediatric surgery. We would like to review our surgical experience and to assess the clinical outcomes and problems encountered during thoracoscopic surgery in children and adolescents focusing on children less than 6 years old. PATIENTS AND METHODS: In 1992 and 2003, 977 patients underwent thoracoscopic surgery for chest diseases. Among them, from 1994 to 2003, 71 children and adolescents underwent thoracoscopic surgery. The thoracoscopic surgical procedures were classified into thoracoscopic surgery (TS) and video-assisted thoracic surgery (VATS). TS was performed through three or four trocar ports. VATS was performed through a small chest incision (minithoracotomy) with one or two trocar ports. RESULTS: There was no morbidity or mortality associated with the thoracoscopic surgical procedures. None of the patients required a conversion to standard thoracotomy. The thoracoscopic surgical procedures were feasible in 71 children and adolescents with chest diseases including spontaneous pneumothorax, pulmonary nodules, diffuse pulmonary disease, pulmonary sequestration, and mediastinal tumors, and in those with palmar hyperhidrosis. TS was performed on two of five children with benign diseases including diffuse pulmonary disease and mediastinal bronchogenic cyst. VATS was performed on three children with pulmonary sequestration, lymphoid interstitial pneumonia and mediastinal Ewing's sarcoma. In one child with lymphoid interstitial pneumonia, the thoracoscopic surgery was converted to VATS because adhesion did not allow TS. The TS allowed rapid histological diagnosis and surgical treatment for benign pulmonary diseases and mediastinal cyst. Patients who were suffering from disease-related symptoms obtained complete relief with an uneventful postoperative course and quickly returned to their normal life. A boy who underwent lobectomy showed a normal growth rate for 4 years postoperatively. In the case of malignant chest diseases, the combination of chemotherapy and VATS was an effective treatment strategy. CONCLUSIONS: We considered that the thoracoscopic approach is a rapid and simple method in the therapeutic diagnosis and surgical treatment of children and adolescents, even in children less than 6 years old. Cosmetic benefits were also obtained for girls. However, the most important consideration is the decision on a treatment strategy made by both pediatrician and thoracic surgeon. PMID- 15834207 TI - Effects of cannabinoids on colonic muscle contractility and tension in guinea pigs. AB - OBJECTIVE: While endogenous cannabinoids regulate various physiologic functions, their role in the intestinal tract is unclear. We continuously recorded colonic motility in conscious guinea pigs. Mechanisms of action then were investigated using guinea pig taenia caecum in vitro. DESIGN: Prospective experimental observations using the cannabinoid agonists 2-arachidonoylglycerol (2-AG) and WIN55212-2; a cannabinoid antagonist, AM281; and ion-channel antagonist. SETTING: University research laboratory. SUBJECTS: Thirty guinea pigs (20 for in vivo study, 10 for in vitro). MEASUREMENTS AND MAIN RESULTS: Colonic motility was monitored in vivo using telemetry via a force transducer attached to the guinea pig taenia caecum. Taenias isolated from other guinea pigs were studied in vitro to assess cannabinoid effects on muscle contractions evoked pharmacologically or electrically. Immediately after cannabinoid injection in conscious guinea pigs, taenial relaxation began peaking at 30 to 40 min. In animals pretreated with AM281, a CB1 cannabinoid receptor antagonist, cannabinoid evoked relaxation was less evident. In vitro, cannabinoids suppressed KCl-induced taenial contractions; this suppression was opposed by charybdotoxin, a Ca(2+)-activated K(+)-channel inhibitor, but not AM281. Cannabinoids decreased amplitude of repeated contractions evoked by electrical stimulation (an effect inhibited by AM281) but not muscle tension. CONCLUSIONS: Cannabinoids decreased intestinal tract tension in vivo, apparently via central CB1 receptors. This differs from peristaltic suppression. PMID- 15834208 TI - Histopathological study of tissue reaction to pacemaker electrodes implanted in the endocardium. AB - Limited information is available about histopathological reactions to the implanted endocardial electrodes of pacemakers (PM). Gross anatomic and histologic studies of tissue reactions to PM electrodes were made in thirteen autopsy cases (nine men and four women, ages 25 approximately 89 years, mean age 71.8) who died two months to twenty-one years after PM implantation. Nine of them had complete atrioventricular (AV) block, three had sick sinus syndrome, and one had bradycardia-tachycardia syndrome. The direct causes of death were not related to their PM. The tip with projecting tines was implanted in the right ventricle in all patients. At the contact area between the electrode and the endocardium, no tissue reaction was observed in one patient with a history of over sixteen years of PM implantation. However, cardiomyocytes under the tip had been replaced by fibrotic tissue in many other patients. In two patients in particular where the electrode had been implanted at the apex of each right ventricle, all cardiomyocytes had disappeared and only fibrotic tissue and adipose tissue were observed under the tip. These findings suggest that mechanical stress caused by attaching the tip tightly damages cardiomyocytes and brings about changes in the pacing thresholds. In three patients, a space was seen between the tip and the endocardium. A fibrous sheath covering the electrode extended to the tip and formed a thick fibrous cap. This non-excitable fibrous cap acted as a virtual electrode and possibly affected the elevation of the threshold in these patients. In four patients, extensive myocardial fibrosis due to disease, e. g. previous myocardial infarction, dilated cardiomyopathy, amyloidosis, or sarcoidosis, was found in the area surrounding the tip and also might affect the elevation of the threshold. We concluded that elevation of pacing thresholds after PM implantation is not due to reactive endocardial thickening. The space between the tip and the endocardium is occupied by a fibrous sheath, and an overly tight attachment damages cardiomyocytes causing replacement fibrosis. Thus, it is not desirable in some patients to insert the electrodes into the apex, where the myocardium is thin. To avoid the elevation of thresholds, development of further devices is necessary to allow electrode fixation to the endocardium with a more suitable pressure level. PMID- 15834209 TI - Pemphigoid nodularis: two case studies and analysis of autoantibodies before and after the development of generalized blistering. AB - We report 45- and 61-year-old women with generalized prurigo nodularis-like eruption whose clinical, histologic and immunopathologic features were consistent with the diagnosis of pemphigoid nodularis. In one case, nodular lesions preceded the onset of generalized blistering by two years and in the other, no definite blister nor erosion was seen except for some appearing on the soles during the course of the disease. Western immunoblotting of EDTA-separated epidermal extracts revealed that the 230-kD bullous pemphigoid (BP) antigen was recognized by circulating autoantibodies in the patient sera, but the 180-kD BP antigen was not. The 180-kD BP antigen was recognized weakly by immunoblotting of the 180-kD BP antigen NC16a domain fusion protein, which shows high detection sensitivity. These findings suggested that weak reactivity of autoantibodies with either whole or a part of the 180-kD BP antigen molecule in some way accounts for negligible or localized blister-formation in this disorder. However, no particular change was noted in the reactivity with 180-kD BP antigen between the patient serum obtained before and after the development of generalized blistering. It is possible that different factors from the changes in serum reactivity with BP antigens may be involved in initiating generalized blistering. PMID- 15834210 TI - Resection of liver metastases from an alpha-fetoprotein-producing gastric cancer. AB - We describe successful resection of rapidly enlarging liver metastases from an alpha-fetoprotein-producing gastric cancer, as these usually carry a dismal prognosis. A 68-year-old woman underwent distal gastrectomy for an alpha fetoprotein-producing gastric cancer without liver metastasis. The tumor was a moderately differentiated tubular adenocarcinoma with invasion of the muscularis propria. Venous and lymphatic invasion were noted, as was metastasis to lymph nodes along the greater curvature. Serum alpha-fetoprotein was 331 ng/mL before gastrectomy, decreasing to 18.6 ng/mL by postoperative day 28. At 2 months after operation, computed tomography detected no metastasis, but at 4 months alpha fetoprotein increased to 2,190 ng/mL, and at 5 months liver tumors were detected by ultrasonography and computed tomography. Serum alpha-fetoprotein increased to 5,673 ng/mL, and serum PIVKA2 concentration was 18 mAU/ml just before operation. Extended left hepatectomy was performed. The resected specimen of segment 4 contained two well-defined tumors, measuring 5 x 4 cm and 2 x 2 cm, while that of segment 5 contained a similar-appearing tumor measuring 2 x 2 cm. The resected tumors had the same histologic appearance as the previously removed gastric cancer. The tumor cells were immunohistochemically reactive for alpha fetoprotein. The postoperative course was uneventful, and the patient was discharged on postoperative day 15. Serum alpha-fetoprotein decreased to 20 ng/mL by postoperative day 15, and to 5 ng/mL by 2 months after operation. For 5 months since operation, no recurrence has become evident, and serum alpha-fetoprotein has remained within the normal range. PMID- 15834212 TI - Ethanol treatment reduces bovine bronchial epithelial cell migration. AB - BACKGROUND: Chronic ethanol abuse is associated with significant lung disease. Excessive alcohol intake increases risk for a variety of respiratory tract diseases, including pneumonia and bronchitis. Damage to airway epithelium is critical to the pathogenesis of airway disorders such as chronic bronchitis and chronic obstructive pulmonary disease. The ability of the airway epithelium to repair itself is an important step in the resolution of airway inflammation and disease. Ethanol exposure is known to modulate signaling systems in bronchial epithelial cells. We hypothesize that chronic ethanol exposure down-regulates the adenosine 3':5'-cyclic monophosphate signaling cascade in airway epithelial cells, resulting in decreased epithelial cell migration and repair. METHODS: We evaluated the effect of ethanol on primary cultures of bovine bronchial epithelial cells in in vitro models of cell migration, wound repair, cell attachment, and cell spreading. RESULTS: Ethanol causes a concentration-dependent effect on closure of mechanical wounds in cell monolayers. Pretreatment of cells with 100 mm ethanol for 24 hr further slows wound closure. Ethanol pretreatment also reduced the protein kinase A response to wounding and made the cells unresponsive to stimuli of protein kinase A that accelerate wound closure. The effects of ethanol on cell migration in wound closure were confirmed in another assay of migration, the Boyden chamber cell migration assay. Prolonged treatment with ethanol also reduced other cell functions, such as spreading and attachment, which are necessary for epithelial repair. CONCLUSIONS: Ethanol modulates signaling systems that are relevant to airway injury and repair, suggesting that chronic, heavy ethanol ingestion has a detrimental impact on airway repair. Impaired response to inflammation and injury may contribute to chronic airway disease. PMID- 15834213 TI - Association between alcoholism and gamma-amino butyric acid alpha2 receptor subtype in a Russian population. AB - BACKGROUND: Two recent large genetic studies in the US population have reported association between genetic variation in gamma-amino butyric acid alpha2 receptor subtype (GABRA2) and risk for alcohol dependence. The goal of this study was to test whether GABRA2 is associated with alcohol dependence in a sample of Russian alcohol-dependent men. METHODS: A total of 113 Russian alcohol-dependent men and 100 male population control subjects were recruited in St. Petersburg and genotyped for seven GABRA2 single-nucleotide polymorphisms (SNPs) using real-time PCR (TaqMan). Six SNPs were located in a GABRA2 haplotype block previously associated with alcohol dependence (AD) in the US population. SNPs and haplotypes were tested for an association to AD using chi analysis and a likelihood ratio based statistic implemented in the software COCAPHASE. RESULTS: Significant associations between two SNPs and AD were observed (p < 0.05). In addition, a trend-level association was observed between AD and three adjacent SNPs (p < 0.1). Associated alleles were carried in a haplotype that was present at frequencies of 0.37 and 0.48 in the control and alcohol-dependent populations, respectively (p < 0.06). Tight linkage disequilibrium spanning from the central portion of the gene to the 3' end was observed in this population. Comparison of the findings to the previously published studies in the US population revealed a highly similar linkage disequilibrium pattern in this population. CONCLUSIONS: These findings suggest that genetic variants of GABRA2 increase risk for AD in the Russian population and provide additional support to the hypothesis that polymorphic variation at the GABRA2 locus plays an important role in predisposing to AD at least in European-ancestry populations. PMID- 15834214 TI - Autonomic modulation of altered diurnal hemodynamic profiles in ethanol-fed hypertensive rats. AB - BACKGROUND: A recent report from the authors' laboratory showed that circadian hemodynamic rhythms are altered in hypertension due partly to irregularities in cardiovascular autonomic control. This study investigated the long-term effects of chronic ethanol feeding (5% w/v, 12 weeks) on the circadian profiles of blood pressure (BP), heart rate (HR), and their variability in spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) rats. METHODS: Radiotelemetry was used for hemodynamic monitoring. The time-domain measures of the variability of BP (standard deviation of mean arterial pressure [SDMAP]) and HR (standard deviation of R-R intervals [SDRR] and root mean square of successive differences in R-R intervals [rMSSD]) were taken as indices of cardiovascular autonomic activity. RESULTS: Control WKY rats displayed normal circadian rhythms (i.e., higher dark-time compared with light-time values) of BP, HR, and their variability indices during weeks 6 through 12 of the study. These circadian rhythms were abolished (BP), inverted (HR, SDMAP), or enhanced (SDRR, rMSSD) in SHRs. Ethanol feeding produced sustained hypotension during both light and dark cycles that was 3-fold greater in SHRs than in WKY rats. SDMAP was not affected by ethanol in WKY rats and showed reductions, mostly during light times, in SHRs. Ethanol also caused significant increases in HR in SHRs only during dark periods, probably because of the inhibition of cardiac vagal activity as indicated by temporal reductions in rMSSD. CONCLUSIONS: These results implicate autonomic factors in the altered diurnal hemodynamic profile by ethanol in SHRs and highlight the possibility of increased risk of cardiac events in hypertensive patients due to alcohol use. PMID- 15834215 TI - Investigating the structure of craving using structural equation modeling in analysis of the obsessive-compulsive drinking scale: a multinational study. AB - BACKGROUND: Currently, there is no agreement among researchers on the definition of craving and its underlying theoretical model. The Obsessive-Compulsive Drinking Scale (OCDS) seems to measure certain aspects of craving, but its theoretical basis remains unclear. The aim of this study was to investigate the structure of alcohol craving, using OCDS data. METHODS: OCDS data from four studies were pooled to obtain a large and heterogeneous sample of 505 participants. All participants were treatment-seeking alcoholics meeting DSM-IV criteria for alcohol dependence. The factor structures of the OCDS previously found were evaluated using confirmatory factor analyses. The goodness of fit of these solutions was compared with those of alternative causal models: an obsessive-compulsive disorder model, an inhibition model, and a cognitive behavioral model. These alternative models were based on modern theories about craving and were tested in the OCDS data, using structural equation modeling. In this way, the current study replaced simple correlational analysis by a more sophisticated causal way of analyzing the underlying structure of the OCDS items. The best fitting model was selected by comparing the mean discrepancy between the implied and observed matrices of the models. RESULTS: The data showed that the previously reported factor structures had to be rejected. Also, the inhibition model and obsessive-compulsive disorder model did not fit the data. The cognitive behavioral model showed encouraging fit. Optimizing strategies were applied to further improve the fit of this model, which resulted in a model with close fit to the data. CONCLUSIONS: The causal cognitive-behavioral model proved to be superior. It showed that the OCDS contains many items that do not represent the core concept of craving but instead are indicators for the consequences of craving. From this model, it seems that craving, in a narrow sense, can be reliably assessed with only five items of the OCDS. PMID- 15834216 TI - Dissection of hypothalamic-pituitary-adrenal axis pathology in 1-month-abstinent alcohol-dependent men, part 1: adrenocortical and pituitary glucocorticoid responsiveness. AB - BACKGROUND: Long-term ingestion of alcohol produces marked alterations in hypothalamic-pituitary-adrenal axis activity. The authors engaged in a series of studies to determine the distinct role of the hypothalamus and the pituitary and adrenal glands in the disturbances observed in abstinent alcohol-dependent subjects. In this first of a two-part study, the authors report on (1) the basal secretory profile of corticotropin and cortisol from 2000 to 0800 hrs, (2) adrenocortical sensitivity in both the presence and absence of endogenous pituitary activation, and (3) pituitary glucocorticoid sensitivity to dexamethasone. METHODS: Eleven male, 4 to 6 weeks abstinent, alcohol-only dependent subjects and 10 age-matched male healthy controls were studied. Basal circulating concentrations of corticotropin and cortisol were obtained from 2000 to 0800 hr. A submaximal dose of cosyntropin (0.01 microg/kg), a corticotropin analogue was then administered to assess adrenocortical sensitivity. In a separate session, cosyntropin was administered following high-dose dexamethasone (8 mg iv) to assess adrenocortical sensitivity in the relative absence of endogenous corticotropin. In addition, the corticotropin response to dexamethasone was measured to determine pituitary glucocorticoid responsiveness. RESULTS: Cortisol, but not corticotropin, pulse amplitude (p < 0.05) and mean concentration (p= 0.05) was significantly lower in alcohol-dependent subjects compared with controls. The cortisol response to cosyntropin was lower in alcohol dependent subjects following endogenous corticotropin suppression by high-dose dexamethasone (p <0.04) but not without dexamethasone pretreatment. Mean corticotropin (p <0.004) and cortisol (p <0.05) concentrations in response to dexamethasone were attenuated in the patients compared to controls. Basal concentrations of 11-deoxycortisol, the precursor to cortisol, were also decreased in alcohol-dependent subjects (p <0.05). CONCLUSION: Attenuated basal and stimulated adrenocortical concentrations in abstinent alcohol-dependent men are coupled with a nonhomeostatic increase in pituitary glucocorticoid inhibition. A decrease in stress-axis responsivity in alcohol dependence may have implications for treatment outcome. PMID- 15834217 TI - Dissection of hypothalamic-pituitary-adrenal axis pathology in 1-month-abstinent alcohol-dependent men, part 2: response to ovine corticotropin-releasing factor and naloxone. AB - BACKGROUND: Pituitary and adrenal responsiveness is suppressed in abstinent alcohol-dependent individuals. To clarify the specific organizational disruption in hypothalamic-pituitary-adrenal functioning during early abstinence, the authors separately assessed each level of the stress-response axis. In this second of a two-part study, ovine corticotropin-releasing factor (oCRH) was used to stimulate the pituitary corticotrophs, and naloxone was used to activate the axis at the hypothalamic level. In addition, pulsatile characteristics of corticotropin and cortisol were assessed over a 12-hr period (0800 to 2000 hr). METHODS: Eleven abstinent alcohol-dependent men and 10 healthy comparison participants were assessed. All participants were between the ages of 30 and 50 years, and alcohol-dependent patients were abstinent from 4 to 6 weeks. Basal concentrations of corticotropin and cortisol were obtained every 10 min from 0800 to 2000 hr and subjected to pulsatile analysis. Plasma corticotropin and cortisol concentrations were then obtained every 5 to 10 min after low-dose, intravenously administered doses of oCRH (0.4 microg/kg) or naloxone (0.125 mg/kg). Medications were administered at 2000 hr and the two challenge studies were separated by 48 hr. RESULTS: Pulsatile analysis revealed that the mean corticotropin amplitude was increased in alcohol-dependent patients relative to controls (p <0.05). Other pulsatile characteristics of corticotropin and all cortisol pulsatile measures were not significantly different between the two groups. The integrated cortisol response to oCRH was significantly lower in alcohol-dependent patients compared with controls (p <0.01), but the integrated corticotropin response was not significantly different. In contrast, neither the corticotropin nor the cortisol response to naloxone was significantly different between groups. CONCLUSIONS: Adrenocorticoid hyposensitivity persists after oCRH infusion for at least 1 month after cessation of drinking, whereas hyporesponsiveness of the pituitary corticotrophs to CRH seems to resolve with continued abstinence. The authors suggest that adrenocortical hyporesponsiveness during prolonged abstinence may impact relapse risk. PMID- 15834218 TI - EEG spectral changes in treatment-naive, actively drinking alcoholics. AB - BACKGROUND: The current study examines the EEG spectra of actively drinking, treatment-naive alcoholics (TxNAs). METHODS: EEGs were gathered on 51 TxNAs and age- and sex-matched controls during closed-eyes conditions. Participants were excluded for lifetime diagnoses of psychiatric or substance abuse disorders. Power for the theta to high beta bands was examined across midline electrodes. RESULTS: The TxNA sample exhibited a nexus of disinhibited traits associated with the vulnerability to alcoholism and had developed alcohol dependence but no other diagnosable psychiatric or substance abuse disorders. The TxNAs evidenced higher power for all EEG bands compared with controls. The magnitude and anterior posterior extent of the group differences varied across bands. Within TxNA, EEG power was negatively correlated with average and peak alcohol drinking duration dose. CONCLUSIONS: Increased EEG power across the theta to high beta bands distinguishes TxNAs without comorbid diagnoses from controls. These effects varied across bands in magnitude and spatial extent, suggesting different effects for the different EEG spectral generators. The authors hypothesize that the increased power in these individuals is a trait difference associated with the inherited nexus of disinhibited traits and its manifestation in alcoholism. Based on the strong negative correlations with alcohol use variables, the authors speculate that decreases in EEG power are a morbid effect of long-term alcohol abuse. They acknowledge that this hypothesized effect of alcohol abuse on EEG power is opposite to the increased EEG power that they hypothesize is associated with alcoholism and its inherited nexus of disinhibited traits. An implication of this model is that with continuing alcohol abuse, the increased EEG power in TxNAs will eventually be overpowered by the effects of long-term severe alcohol abuse. This model predicts that in very long-term alcoholics, EEG power would be equal to or lower than that of age- and sex-comparable controls. PMID- 15834219 TI - Alcohol dose-dependent increases in smoking urge in light smokers. AB - BACKGROUND: The current study assessed dose-dependent effects of alcohol compared with placebo on ratings of urge to smoke in light smokers. METHODS: Sixteen nonalcoholic social drinker-smokers were tested individually in three separate early evening sessions where they received a placebo (with 1% ethanol as a taste mask), a low-dose (0.4 g/kg) alcoholic beverage, or high-dose (0.8 g/kg) alcoholic beverage administered in random order. Participants refrained from smoking 2 hr before and throughout the entire early evening experimental sessions. Two subfactors of the Brief Questionnaire of Smoking Urges, BQSU; (factor 1, urge to smoke for stimulation; factor 2, urge to smoke to relieve negative mood and withdrawal) were assessed at baseline and again at rising and declining portions of the blood alcohol curve. RESULTS: Both the high and low doses of alcohol significantly increased BQSU factor 1 scores during the rising and declining blood alcohol concentration (BAC) limbs (p < 0.05). Comparisons across doses during both limbs revealed that the high dose significantly increased factor 1 smoking urge compared with the low dose and placebo beverage (p < 0.05, high > low = placebo). Alcohol tended to increase factor 2 scores throughout the BAC curve, but levels were not as increased as factor 1 scores. Finally, there was no significant association between participants' smoking levels and smoking urge ratings during the high- and low-dose sessions. CONCLUSIONS: The results support a dose-dependent alcohol-induced increase in smoking urge in cigarette-deprived light smokers. These smoking urge increases were apparent during the rising limb of the BAC and maintained throughout the declining limb. Smoking urge increases were greater for positive reinforcing effects than for negative reinforcing effects. PMID- 15834220 TI - Modulation of ethanol withdrawal-induced anxiety-like behavior during later withdrawals by treatment of early withdrawals with benzodiazepine/gamma aminobutyric acid ligands. AB - BACKGROUND: Anxiety states, including those arising during acute or protracted withdrawal periods, may be precipitating factors in alcoholic relapse. Given the cyclical nature of ethanol withdrawal associated with repeated cycles of ethanol intake and abstinence in a pattern that often spans years, meaningful attempts to model ethanol withdrawal-associated anxiety should incorporate cycled ethanol treatments. The studies reported herein examined the effects of gamma aminobutyric acid-modulating drugs on social interaction behavior-an established model of anxiety-in rats exposed to repeated cycles of ethanol treatment and withdrawal. METHODS: Rats were exposed to 8 to 12 g/kg/day ethanol during three 7 day dietary cycles (5 days on ethanol diet followed by 2 days on control diet). Ethanol was administered either at hour 4 of withdrawal after cessation of each of the first 2 ethanol cycles or during the final withdrawal only. In other groups, the early withdrawals were treated with alphaxalone, diazepam, PK11159, or flumazenil to block anxiety-like behavior during an untreated later (third) withdrawal. The benzodiazepine inverse agonist DMCM (methyl-6, 7-dymerhoxy-4 ethyl-beta-carboline-3-carboxylate) was also given repeatedly to determine whether it would sensitize anxiety-like behavior during a future withdrawal. Finally, the effects of all drugs on deficits in locomotor behavior were assessed. RESULTS: Pretreatment of earlier withdrawals with alphaxalone, diazepam, ethanol, or flumazenil reduced social interaction deficits during a later withdrawal, but pretreatment with PK11195 did not. In contrast, DMCM administered in lieu of early withdrawals increased social interaction deficits during an untreated later withdrawal. Locomotor deficits were significantly reversed only by the acute ethanol and diazepam treatment during the final withdrawal. CONCLUSIONS: Single-dose administration of drugs that enhance or diminish activity at benzodiazepine-gamma-aminobutyric acid- receptors during earlier withdrawals reduced or potentiated, respectively, anxiety-like behavior during later, drug-free withdrawals. These results support the potential of the novel strategy of using prophylactic therapy administered during early withdrawals to ameliorate symptoms of later withdrawals. PMID- 15834221 TI - Role of the serotonin transporter gene and family function in adolescent alcohol consumption. AB - BACKGROUND: That the extent to which a particular individual will engage in problematic behaviors such as delinquency, violence, or drug abuse is determined by the way psychosocial, situational, and hereditary factors interact is widely accepted. However, only recently have researchers begun to investigate the interactions between specific genotypes and psychosocial factors in relation to behavior. The purpose of the present study was to investigate possible interactions between a polymorphism in the promoter region of the serotonin transporter (5-HTT) gene and family relations on adolescent alcohol consumption. METHODS: A cross-sectional study with a randomized sample from a total population of 16- and 19-year-old adolescents from a Swedish county was conducted. Eighty one male and 119 female adolescents, who volunteered to participate after having answered a questionnaire, were randomly selected from quartiles of volunteers representing various degrees of psychosocial risk behavior. RESULTS: 5-HTT genotype (p=0.029) and family relations (p=0.022) predicted alcohol consumption independently as well as through an interaction with one another (p=0.05). The model explained 11% of the variance in alcohol consumption. In a binary logistic model, we found that adolescents with the LS variant of the 5-HTT gene and with family relations being "neutral" or "bad" had a 12- to 14-fold increased risk for high intoxication frequency. CONCLUSIONS: In sum, our results show that a functional polymorphism of the 5-HTT genotype, family relations, and interactions between these variables predict adolescent alcohol consumption in a randomized sample of adolescents. PMID- 15834222 TI - Brain stimulation reward performance and sucrose maintained behaviors in alcohol preferring and -nonpreferring rats. AB - BACKGROUND: The relation between ethanol (EtOH) preference and sensitivity to brain stimulation reward (BSR) was examined under multiples schedules of reinforcement in the current study. For comparison, the study also examined the relation between EtOH preference and motivation for a sweet, palatable sucrose solution under similar schedules of reinforcement. METHODS: To investigate sensitivity to BSR performance, alcohol-preferring and -nonpreferring rats were tested using the curve-shift (rate-frequency) paradigm under several intensity levels during a 20-min session. Animals were first trained under an optimal current intensity, which produced maximal responding (i.e., 100%) across a series of descending frequencies (i.e., 300-20 Hz). BSR was then evaluated at 100%, 75%, and 50% of the optimal current. The sensitivity of the curve-shift method was further evaluated under the animal's optimal current using the FR1, FR6, and FR12 schedules. To examine responding for the sucrose solution, a separate group of alcohol-preferring and -nonpreferring rats was initially stabilized on an FR1 schedule and then subsequently on FR6 and FR12 schedules. RESULTS: The results demonstrated that reducing the reinforcing efficacy of BSR via reduction in current intensity/reinforcement schedule produced marked orderly rightward shifts in the rate-frequency curves relating responding to stimulation frequency in both rat lines. However, no differences were found between the lines with either manipulation. Specifically, both lines demonstrated orderly reductions in response rate and increases in BSR threshold parameters (i.e., half maximal frequency/responding, minimum and maximum frequencies). In contrast to BSR, genetic selection for EtOH preference was highly associated with responding for the sweet, palatable sucrose solution. The association was even more salient as the reinforcement schedule increased (i.e., reward cost). CONCLUSION: The results demonstrate that responding for BSR is not associated with EtOH preference, insofar as alcohol-preferring and -nonpreferring rats respond similarly under an array of reinforcement schedules and current intensities. In contrast, genetic selection for EtOH preference is highly associated with responding for a palatable sucrose reward, and the relation increases as the reward cost for the sucrose increases. These findings suggest that similar/overlapping mechanisms of action regulate the reinforcing properties of EtOH and sucrose but that overlapping yet distinct neuronal mechanism may modulate the reward characteristics of BSR and EtOH preference. PMID- 15834223 TI - Effects of neuropeptide Y on appetitive and consummatory behaviors associated with alcohol drinking in wistar rats with a history of ethanol exposure. AB - BACKGROUND: Neuropeptide Y (NPY) reduces ethanol intake under free access conditions in Wistar rats with a history of prolonged ethanol vapor exposure. The current study was designed to determine whether NPY differentially alters ethanol associated appetitive behavior (i.e., lever pressing) or ethanol consumption in Wistar rats with a history of ethanol vapor exposure. METHODS: Wistar rats were first trained to self-administer 10% ethanol in a paradigm that provided 25 min of free access to 10% ethanol after completing a 20-lever press response requirement (i.e., an RR20 schedule). After stable level lever pressing was established, operant sessions were suspended during a 9-week period of ethanol vapor exposure. Self-administration sessions were then reinstituted, and a fixed time (FT) schedule of 10% ethanol access was used to assess the effects of ethanol exposure and NPY on lever pressing and drinking behavior. Under the FT schedule, the maximum number of lever presses emitted within 10 min was assessed before providing access to 10% ethanol. RESULTS: Ethanol vapor exposure did not alter patterns of lever pressing under the RR20 schedule, but lever presses emitted under the FT schedule were reduced after ethanol vapor exposure. Ethanol intake was significantly increased after ethanol vapor exposure. NPY significantly reduced ethanol intake but did not significantly reduce lever pressing under the FT schedule. CONCLUSIONS: Taken together, these data suggest that chronic ethanol exposure increases ethanol intake without clearly enhancing its reinforcing value. Furthermore, NPY has a greater impact on the consummatory factors mediating ethanol intake than appetitive factors mediating ethanol seeking. PMID- 15834224 TI - Short and prolonged periods of maternal separation and voluntary ethanol intake in male and female ethanol-preferring AA and ethanol-avoiding ANA rats. AB - BACKGROUND: Genetic as well as environmental factors can affect the propensity for psychopathology and/or drug dependence. Maternal separation represents an animal experimental model that is useful in studies of effects of early life experiences. The authors have established a protocol for short and prolonged periods of maternal separation to study adult neurochemistry, behavior, and ethanol intake and have previously reported alterations in ethanol intake in Wistar rats and ethanol-preferring rats. The aim of the current study was to more thoroughly study how early life experiences affect an inherited propensity for high and low ethanol intake, respectively, in male and female ethanol-preferring AA (Alko alcohol) and ethanol-avoiding ANA (Alko, Non-Alcohol) rats. METHODS: AA and ANA pups were assigned to one of three different rearing conditions: 15 min (MS15) or 360 min (MS360) of daily maternal separation in litters or normal animal facility rearing (AFR) during postnatal days 1 to 21. In adulthood, voluntary ethanol intake was investigated using the two-bottle free choice paradigm. RESULTS: In male ethanol-preferring AA rats, MS15 resulted in a lower intake and fewer high-preferring animals at 8% and 10% ethanol compared with MS360 rats. The male MS360 rats had a higher ethanol intake at 8% and 10% ethanol in comparison with AFR rats. In contrast, the female AA MS15 and MS360 rats had a lower ethanol intake and a lower preference for the 10% ethanol solution compared with the female AA AFR rats. In male and female ANA rats, no major separation induced effects were found. CONCLUSIONS: The current results show that genetic inheritance can be affected by environmental manipulations in AA rats with an inherent high ethanol intake. The findings in female ethanol-preferring AA rats give further evidence of a differential outcome of maternal separation in male and female rats, as previously shown. PMID- 15834225 TI - New approach to the validity of the alcohol use disorders identification test: stratum-specific likelihood ratios analysis. AB - BACKGROUND: Previous validation studies of the Alcohol Use Disorders Identification Test (AUDIT) used either the fixed-threshold approach or receiver operating characteristic (ROC) analyses. This study was aimed to conduct a stratum-specific likelihood ratio (SSLR) analysis on the validity of the AUDIT for detecting alcohol use disorders. METHODS: A two-phase validation study was conducted among nonpsychiatric inpatients at a general hospital, using the AUDIT for phase 1 screening and the Schedules for Clinical Assessment in Neuropsychiatry for the phase 2 interview. All of the patients with positive screening results and a subsample of the patients with negative screening results were interviewed in the second phase. The validity of the AUDIT for current alcohol use disorders was estimated using the ROC first, and an SSLR analysis was performed subsequently. RESULTS: In 422 inpatients who completed the screening, 193 had a negative result with a zero score, 103 had a negative result with a nonzero score, and 126 had a positive result. Twenty screened patients with a negative result and a nonzero score and 107 patients with a positive result were interviewed in the second phase, in which 59 patients were found to have current alcohol use disorders according to DSM-IV criteria. The area under the ROC curve of the AUDIT for current alcohol use disorders was 0.98. The sensitivity and specificity of the AUDIT at the optimal cutoff of 8 were 0.97 and 0.90, respectively. Three strata of the AUDIT scores (0-7, 8-13, and > or = 14) were derived for alcohol use disorders, with the SSLRs being 0.04, 3.67, and 49.72, and the corresponding posttest probabilities being 0.01, 0.42, and 0.91, respectively. CONCLUSIONS: The AUDIT is a valid screening tool for alcohol use disorders, and the three strata derived from the SSLR analysis are informative and readily applicable in clinical practice. PMID- 15834226 TI - Impact of a workplace peer-focused substance abuse prevention and early intervention program. AB - BACKGROUND: PeerCare is a workplace peer intervention program that focuses on changing workplace attitudes toward on-the-job substance use and trains workers to recognize, intervene with, and refer coworkers who have a problem. METHODS: Monthly injuries at the study company (January 1983 through June 1996) were compared to counts at four other companies in the same industry. Using these panel data, fixed-effects negative binomial regression measured the association of the percentage of the workforce covered by PeerCare with the workplace injury rate. RESULTS: For every 1% increase in the workforce covered with PeerCare, the risk of injury declined by 0.9984 (95% confidence interval, 0.9975-0.9994). These findings suggest that, by June 1996, when 86% of the workforce was covered under PeerCare, the program had reduced injury rates by an average 14% per month. CONCLUSIONS: The findings support the implementation of peer intervention programs as a means to reduce workplace injuries. PMID- 15834227 TI - The 8-year course of alcohol abuse: gender differences in social context and coping. AB - BACKGROUND: The aim of this study was to compare women (n=230) and men (n=236) who had alcohol use disorders in terms of social context and coping methods and in terms of changes in these indices associated with participation in professional treatment and Alcoholics Anonymous (AA). METHODS: Initially untreated problem drinkers were followed up for 8 years. RESULTS: Women and men did not differ in regard to the type of help received, but women had longer professional treatment. At baseline, women had more stressors and fewer resources from family and relied more on avoidance coping and drinking to cope. During the next 8 years, women, more so than men, increased on approach coping and reduced their use of avoidance coping and drinking to cope. When baseline status was controlled, women had better social resource, coping, and drinking outcomes than men did at 1 year and 8 years. A longer duration of professional treatment during year 1 was associated with improved approach coping among men but not women. A longer duration of AA attendance during year 1 and the full 8 years was associated with more resources from friends, more use of approach coping, and less drinking to cope. In turn, more friends resources and approach coping and less drinking to cope were associated with better drinking outcomes. Decreases in avoidance coping and drinking to cope were more strongly associated with better drinking outcomes among men than among women. CONCLUSIONS: It may be important to target men for formal services or self-help to increase their use of approach coping in efforts to maintain abstinence. Women's strategies for improving their social context need further explication to be adapted for transfer to male problem drinkers. PMID- 15834228 TI - Epidemiological evidence that acetaldehyde plays a significant role in the development of decreased serum folate concentration and elevated mean corpuscular volume in alcohol drinkers. AB - BACKGROUND: Elevated mean corpuscular volume (MCV) is a traditional biological marker for alcohol abuse and alcoholism, but the underlying mechanism is unclear. Three recent epidemiologic studies consistently showed that MCV was elevated by alcohol drinking more markedly among individuals with genetically inactive aldehyde dehydrogenase-2 (ALDH2) (encoded by ALDH2*2 mutant allele) than those with active ALDH2 (encoded by ALDH2*1/2*1 genotype), suggesting that the elevated MCV was etiologically linked to acetaldehyde exposure. The purpose of the present study was to clarify further this relationship by examining the status of folate and vitamin B12. METHODS: The study participants were 159 men who were aged 40 to 69 years and randomly selected from a Japanese rural population. The genetic polymorphism of ALDH2 was determined by PCR-restriction fragment length polymorphism method; data on alcohol drinking and other lifestyles were collected using a structured questionnaire; serum concentrations of folate and vitamin B12 were measured using the protein competitive reaction method, and blood cell counts were measured by routine methods. A multiple linear regression model was used to analyze the data. RESULTS: : The relationship between alcohol drinking and serum folate concentration was significantly different between ALDH2 genotypes, indicating that the reduction of serum folate by alcohol drinking was more marked in men with ALDH2*1/2*2 than those with ALDH2*1/2*1. The relationship between alcohol drinking and elevated MCV was significantly stronger in men with ALDH2*1/2*2 than those with ALDH2*1/2*1 even after adjustment for serum folate and vitamin B12 concentrations. CONCLUSIONS: These findings indicate that acetaldehyde plays a significant role in the development of decreased serum folate concentration and elevated MCV by alcohol drinking. PMID- 15834229 TI - College students lack knowledge of standard drink volumes: implications for definitions of risky drinking based on survey data. AB - BACKGROUND: College students tend to pour single servings of beer and liquor that are larger than commonly used standards. The reasons for this are unknown. Students might overpour because they lack knowledge of standard serving sizes. Alternatively, they might know how much alcohol to pour but simply have difficulty pouring the correct amounts. Misperceptions of standard serving sizes could lead to inaccuracies in self-reported consumption. If this is the case, then the validity of students' responses on alcohol surveys and the definitions of risky drinking that are based on them would be called into question. This study examined how college students define standard drinks, whether their definitions are similar to the definitions commonly used by alcohol researchers and government agencies, and whether their definitions of standard drinks are related to the sizes of the drinks that they pour. The study also examined whether feedback regarding the accuracy of their definitions of standard drinks leads students to alter their self-reported levels of consumption. METHODS: Students (N = 133) completed an alcohol survey and performed tasks that required them to free-pour a single beer, glass of wine, shot of liquor, or the amount of liquor in a mixed drink. Roughly half of the students received feedback regarding their definitions of standard drinks. All students then were resurveyed about their recent levels of consumption. RESULTS: With the exception of beer, students incorrectly defined the volumes of standard servings of alcohol, overestimating the appropriate volumes. They also overestimated appropriate volumes when asked to free-pour drinks. Positive relationships existed between students' definitions of standard drinks and the sizes of the drinks that they free-poured. Feedback regarding misperceptions of standard drink volumes led to an increase in levels of self-reported consumption, suggesting that students' original estimates of their alcohol consumption were too low. CONCLUSIONS: Despite the recent focus on alcohol education and prevention at the college level, college students have not been taught how to define standard drinks accurately. They tend to overstate the appropriate volumes, leading them to overpour drinks and underreport levels of consumption. Self-reported consumption levels are altered by feedback regarding the accuracy of students' definitions of standard drinks. The findings raise important questions about the validity of students' responses on alcohol surveys and the definitions of risky drinking that are based them. PMID- 15834230 TI - Brainstem norepinephrine neurons mediate ethanol-evoked pressor response but not baroreflex dysfunction. AB - BACKGROUND: Ethanol elicits strain-dependent blood pressure and baroreflex sensitivity responses in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) rats; the mechanisms underlying these divergent effects are not clear. The authors tested the hypothesis that differential neuronal actions of ethanol may account for these strain-dependent responses. To this end, the authors investigated the direct effects of ethanol on norepinephrine (NE)-containing neurons in the rostral ventrolateral medulla (RVLM), which modulate sympathetic neuronal activity, and on c-Jun-expressing neurons in the nucleus tractus solitarius (NTS), whose activity is inversely correlated with baroreflex sensitivity. METHODS: In a newly developed model system in conscious, freely moving rats, the effect of intra-RVLM or intra-NTS ethanol was investigated on neuronal NE at the microinjection site (in vivo electrochemistry), blood pressure, heart rate, spontaneous baroreflex sensitivity, and c-Jun expression in the NTS. RESULTS: Ethanol (1, 5, or 10 microg) microinjection into the RVLM elicited dose-dependent increases in RVLM NE and blood pressure in SHRs but not in WKY rats. Ethanol had no effect on the activity of the NE-containing neurons in the NTS of either strain. However, baroreflex dysfunction elicited by intra NTS ethanol in conscious WKY rats was associated with enhanced expression of c Jun in the NTS. CONCLUSIONS: (1) Ethanol activation of the NE-containing neurons in the RVLM of SHRs contributes to the centrally mediated pressor response, (2) the NE-containing neurons in the NTS are not involved in ethanol-induced baroreflex dysfunction, and (3) direct activation of the c-Jun-containing neurons in the NTS is implicated in baroreflex dysfunction elicited by ethanol in normotensive rats. PMID- 15834231 TI - Vasoactive intestinal peptide and corticotropin-releasing hormone increase beta endorphin release and proopiomelanocortin messenger RNA levels in primary cultures of hypothalamic cells: effects of acute and chronic ethanol treatment. AB - BACKGROUND: beta-Endorphin (beta-EP) neurons are involved in ethanol's action on a variety of brain functions, including positive reinforcement. These neurons are innervated by vasoactive intestinal peptide (VIP)-containing and corticotropin releasing hormone (CRH)-containing neurons in the hypothalamus. Whether these neuropeptides affect beta-EP neuronal function in the presence or absence of ethanol has not previously been determined. METHODS: The authors determined the effects of VIP and CRH on gene expression and peptide release from beta-EP neurons in primary cultures of mediobasal hypothalamic cells. The effects of receptor antagonists on VIP- and CRH-induced beta-EP release was determined. Furthermore, the authors studied the effects of acute and chronic treatment with ethanol on the response of beta-EP neurons to VIP and CRH. Real-time reverse transcription polymerase chain reaction was used for messenger RNA (mRNA) detection, and radioimmunoassay was used for hormone measurements. RESULTS: We show that beta-EP neurons responded concentration dependently to VIP and CRH treatments by increasing both beta-EP release and proopiomelanocortin mRNA expression. Simultaneous treatment with a nonspecific receptor antagonist reduced the ability of CRH or VIP to induce beta-EP release from mediobasal hypothalamic cells. Acute treatment with ethanol increased beta-EP neuronal gene expression and the secretory response to CRH and VIP. However, previous exposure to chronic ethanol reduced the CRH and VIP responses of these neurons. CONCLUSIONS: These results indicate that VIP and CRH stimulate beta-EP release from hypothalamic cells in primary cultures and that the stimulatory and adaptive responses of beta EP neurons to ethanol may involve alteration in the responsiveness of beta-EP secreting neurons to CRH and VIP. PMID- 15834232 TI - Associations between alcohol intake and brain volumes in male and female moderate drinkers. AB - BACKGROUND: Alcohol-dependent individuals have brain volume loss. Possibly, moderate drinkers who are not alcohol dependent have similar but less prominent brain damage. The authors investigated whether current or lifetime alcohol intake is related to volumes of total brain, cerebellum, ventricles, peripheral cerebrospinal fluid, and cerebral gray and white matter in moderate drinkers. METHODS: The relation between current or lifetime alcohol intake and brain volumes of 47 male moderate drinkers (current alcohol intake 20 drinks per week, lifetime alcohol intake 240 kg) and 44 female moderate drinkers (current alcohol intake 15 drinks per week, lifetime alcohol intake 170 kg), all without a personal or family history of alcohol dependence, was determined using high resolution magnetic resonance images, corrected for intracranial volume, age, and sex. RESULTS: In males, mean lifetime alcohol intake was positively associated with cerebral white matter volume, particularly in the frontal region. In females, mean lifetime alcohol intake was not associated with brain volumes. Current alcohol intake was unrelated to brain volumes in either males or females. CONCLUSIONS: Neither current nor lifetime alcohol intake is associated with decreases in brain volumes in male or female moderate drinkers. Because all participants had a negative personal and family history of alcohol dependence, the current results relatively purely concern the effects of moderate alcohol intake on brain volumes. PMID- 15834233 TI - Effect of red wine on the intestinal absorption of thiamine and folate in the rat: comparison with the effect of ethanol alone. AB - BACKGROUND: This work aimed to investigate, in the rat, the acute in vitro effect of red wine and the effect of chronic red wine ingestion on the intestinal absorption of thiamine and folate and to compare them with the effects of ethanol alone. METHODS: The effects of red wine and of an ethanol solution (same ethanol concentration as that in the red wine, i.e., 12% [v/v]) on rat jejunal apparent permeability (Papp) to H-thiamine and H-folate in the mucosal-to-serosal direction were investigated. Red wine and ethanol were tested both chronically (21-day consumption) and acutely in vitro. RESULTS: Acutely, both red wine and ethanol 12% (v/v) (both diluted 1:5) reduced (to 65 and 60% of control, respectively) the mucosal-to-serosal Papp to H-thiamine across rat jejunum. Chronic (21-day) ethanol (12% [v/v]) consumption also decreased the Papp to H thiamine (to 33% of control), but red wine consumption for the same period did not change it. Mucosal-to-serosal Papp to H-folate across rat jejunum was not changed by chronic ingestion of red wine or ethanol. Similarly, it was not affected by acute exposition of the tissue to red wine or ethanol. Acute ethanol (0.05% [v/v]) did not affect the Papp to H-thiamine or H-folate in jejunal tissues obtained from control and red wine-treated rats, but it significantly increased the Papp to both H-thiamine and H-folate (to 183 and 197% of control, respectively) in tissues from chronically ethanol-treated rats. CONCLUSIONS: Acute and chronic red wine or ethanol had no effect on the intestinal absorption of folate. However, ethanol, both acutely and chronically, decreased the jejunal absorption of thiamine, and red wine reduced the jejunal absorption of thiamine, but only when tested acutely. These findings show that it is not correct to extrapolate from results obtained with ethanol alone on intestinal permeability to the effect of alcoholic beverage consumption. PMID- 15834235 TI - Early postnatal exposure to alcohol reduces the number of neurons in the occipital but not the parietal cortex of the rat. AB - BACKGROUND: The rat brain undergoes a period of rapid growth in the early postnatal period. During this time, the neocortex seems to be vulnerable to ethanol injury. Subdivisions of the neocortex develop in a temporospatial gradient that is likely to determine their vulnerability to ethanol-induced damage and whether damage is permanent. Therefore, the authors investigated the effect of postnatal ethanol exposure on the neocortex and specific subregions at the cessation of exposure and in the mature brain. METHODS: Four-day-old rat pups with intragastric cannulae were artificially reared from postnatal day (PN) 4 through PN9. Of 12 daily feeds, two consecutive feeds contained either ethanol (4.5 g/kg) or an isocaloric maltose/dextrin solution. On PN10 or PN115, animals were perfused intracardially, and the brains were removed. Stereological methods were used to determine the total number of neurons and glial cells in, and the volume of, the neocortex, the parietal cortex, and the occipital cortex. RESULTS: Exposure to ethanol did not affect body or brain weight at PN10. In contrast, at PN115 forebrain weight was significantly lower in ethanol-exposed animals compared with control-treated animals. There was no effect of treatment on body weight at PN115. On PN10, neocortical volume was 15% smaller in the ethanol exposed animals compared with controls, with no change in the total number of neurons or glial cells. Occipital cortical volume was reduced by 22% in the ethanol-exposed animals, with a significant deficit in the total number of neurons (ethanol-exposed, 2.62 x 10; gastrostomy control, 3.20 x 10). There was no effect of ethanol exposure on the total number of glial cells in the occipital cortex or on any parameter in the parietal cortex. There was also no significant effect of ethanol exposure on the occipital cortex on PN115. CONCLUSIONS: These findings provide support for the hypothesis that a specific area or cell population might be differentially vulnerable to ethanol exposure during the brain growth spurt and that cell deficits evident on PN10 may not be permanent. PMID- 15834234 TI - Acute ethanol inhibits extracellular signal-regulated kinase, protein kinase B, and adenosine 3':5'-cyclic monophosphate response element binding protein activity in an age- and brain region-specific manner. AB - BACKGROUND: As little as a single episode of exposure of the developing brain to ethanol can result in developmental neuropathology and mental retardation. Extracellular signal-regulated kinases (ERKs), protein kinase B (PKB), and adenosine 3':5'-cyclic monophosphate response element binding protein (CREB) are messenger molecules that play important roles in neuronal plasticity and survival. This study was undertaken to examine the effects of acute ethanol on ERK, PKB, and CREB activation in the brain. METHODS: Immunoblot analysis was used to determine the effects of a 1-hr exposure of ethanol on levels of phospho-ERC in primary cortical cultures and in the cerebral cortex, hippocampus, and cerebellum of postnatal day 5 (PN5), postnatal day 21 (PN21), and adult rats. RESULTS: In cortical cultures, ethanol (100 mM) significantly reduced activity dependent activation of phospho-ERK, phospho-PKB, and phospho-CREB by approximately 50%. In PN5 rats, ethanol (3.5 g/kg) inhibited both phospho-ERK and phospho-PKB in the cerebral cortex and hippocampus but was without effect in the cerebellum. A similar brain region-specific inhibition of phospho-ERK was observed in PN21 rats, whereas in adult rats, ethanol inhibited phospho-ERK in all three brain regions. In contrast, ethanol had no effect on phospho-PKB in either PN21 or adult rats. Without exception, ethanol inhibited phospho-CREB in an identical brain region- and age-dependent manner as was observed for phospho ERK. Finally, administration of the NMDA antagonist MK-801 (0.5 mg/kg) to PN5 rats had no effect on phospho-ERK or phospho-PKB levels in any brain region. CONCLUSION: The results demonstrate that acute ethanol inhibits ERK/PKB/CREB signaling in brain. This inhibition occurs in an age- and brain region-specific manner, with inhibition of PKB restricted to a time during the brain growth-spurt period. Furthermore, the lack of effect of MK-801 suggests that inhibition of NMDA receptors is unlikely to play a major role in binge ethanol inhibition of ERK/PKB/CREB signaling in vivo. PMID- 15834237 TI - Ethanol injections in Drosophila. PMID- 15834238 TI - Ayurveda and yoga in cardiovascular diseases. AB - Ayurveda is derived from 2 Sanskrit words, namely, "Ayus" and "Veda," meaning life and knowledge, respectively. It literally means science of life. Ayurveda, of which yoga is an integral part, is widely practiced in India and is gaining acceptance in many countries around the world. It is a comprehensive and a holistic system, the focus of which is on the body, mind, and consciousness. The Ayurvedic treatment consists of the use herbal preparations, diet, yoga, meditation, and other practices. Based on the review of available studies, the evidence is not convincing that any Ayurvedic herbal treatment is effective in the treatment of heart disease or hypertension. However, the use of certain spices and herbs such as garlic and turmeric in an overall healthy diet is appropriate. Many herbs used by Ayurvedic practitioners show promise and could be appropriate for larger randomized trials. Yoga, an integral part of Ayurveda, has been shown to be useful to patients with heart disease and hypertension. Yoga reduces anxiety, promotes well-being, and improves quality of life. Its safety profile is excellent. Its use as a complementary therapeutic regimen under medical supervision is appropriate and could be worth considering. PMID- 15834240 TI - Attitudes about genetic testing and genetic testing intentions in African American women at increased risk for hereditary breast cancer. AB - PURPOSE: To evaluate attitudes about the benefits, limitations, and risks of genetic testing for BRCA1 and BRCA2 (BRCA1/2) mutations and explore testing intentions in African American women at increased risk for hereditary breast cancer. METHODS: Attitudes and intentions were evaluated by telephone in African American women (n = 74) at moderate and high risk for having a BRCA1/2 mutation. RESULTS: Attitudes about the benefits of genetic testing were endorsed at a higher rate relative to limitations and risks; however, only 30% of respondents indicated that they would definitely have testing. In regression analysis, women most likely to be considering testing were those with fatalistic beliefs about cancer and those who believed they had a BRCA1/2 mutation. Women who had two or more affected relatives were also most likely to be considering testing. Women who had a personal history of cancer and those who believed they were at high risk for developing breast cancer were most likely to report greater limitations and risks. Pros scores were higher among women older than age 50 and those who were unemployed. CONCLUSION: Although African American women at moderate and high risk for BRCA1/2 mutations report favorable attitudes about genetic testing, interest in testing may be limited. Women affected with cancer and those who believe they are at a higher risk for developing breast cancer may be most concerned about the negative consequences of testing. Increased attention may need to be given to beliefs about genetic testing and testing motivations during genetic counseling with African American women. PMID- 15834239 TI - Use of an educational computer program before genetic counseling for breast cancer susceptibility: effects on duration and content of counseling sessions. AB - PURPOSE: Patients seeking genetic testing for inherited breast cancer risk are typically educated by genetic counselors; however, the growing demand for cancer genetic testing will likely exceed the availability of counselors trained in this area. We compared the effectiveness of counseling alone versus counseling preceded by use of a computer-based decision aid among women referred to genetic counseling for a family or personal history of breast cancer. METHODS: We developed and evaluated an interactive computer program that educates women about breast cancer, heredity, and genetic testing. Between May 2000 and September 2002, women at six study sites were randomized into either: Counselor Group (n = 105), who received standard genetic counseling, or Computer Group (n = 106), who used the interactive computer program before counseling. Clients and counselors both evaluated the effectiveness of counseling sessions, and counselors completed additional measures for the Computer Group. Counselors also recorded the duration of each session. RESULTS: Baseline characteristics did not differ significantly between groups. Participants and counselors both rated the counseling sessions as highly effective, whether or not the sessions were preceded by computer use. Computer use resulted in significantly shorter counseling sessions among women at low risk for carrying BRCA1/2 mutations. In approximately half of the sessions preceded by clients' computer use, counselors indicated that clients' use of the computer program affected the way they used the time, shifting the focus away from basic education toward personal risk and decision-making. CONCLUSION: This study shows that the interactive computer program "Breast Cancer Risk and Genetic Testing" is a valuable adjunct to genetic counseling. Its use before counseling can shorten counseling sessions and allow counselors to focus more on the clients' individual risks and specific psychological concerns. As the demand for counseling services increases, a program such as this can play a valuable role in enhancing counseling efficiency. PMID- 15834241 TI - What's the message? Interpretation of an uninformative BRCA1/2 test result for women at risk of familial breast cancer. AB - PURPOSE: To test the "false-reassurance hypothesis," which suggests that women who receive an uninformative BRCA1/2 test result may incorrectly conclude that they no longer have an elevated risk, with possible harmful consequences for adherence to breast surveillance guidelines. METHODS: A prospective questionnaire design was used to compare 183 women with an uninformative BRCA test result (94 affected and 89 unaffected) with 41 proven BRCA mutation-carriers and 49 true negatives before and after BRCA1/2 test disclosure. RESULTS: After DNA-test disclosure, test applicants differed from each other with regard to their perception of the likelihood of carrying a deleterious gene (P < 0.0001). The BRCA mutation carriers reported the highest perceived likelihood and the true negatives reported the lowest. Compared to the predisclosure measures, women who received an uninformative DNA test result reported a lower perceived risk after disclosure (P < 0.0001), suggesting a relatively high level of reassurance because of the test result. However, after DNA-test disclosure, only 12 women concluded that the risk of carrying a mutation was nonexistent, and perceived likelihood was significantly associated with the pedigree-based risk assessment (P = 0.0001). Moreover, despite the significant decrease in perceived likelihood for uninformative women, intention to obtain mammograms did not change (P = 0.71); it remained at the same almost optimal level as for BRCA mutation carriers. CONCLUSION: No support was found for the suggestion that the nature of uninformative test results is often misunderstood. Moreover, an uninformative test result did not affect the positive mammography intentions of both affected and unaffected women. PMID- 15834242 TI - Fragile X syndrome carrier screening in the prenatal genetic counseling setting. AB - PURPOSE: To document our experience with fragile X carrier screening. METHODS: In this study, 29,103 women with no known or suspected family history of fragile X syndrome were offered fragile X carrier screening during their prenatal genetic counseling visit. Screening acceptance was analyzed by referral indication, carrier frequencies documented, and prenatal outcome data presented. RESULTS: Overall, 7.9% accepted carrier screening. The premutation frequency was 1 in 382, and the intermediate allele frequency was 1 in 143. CONCLUSIONS: Fragile X screening is a desirable option for some women seeking prenatal genetic counseling and should be made available to this population. PMID- 15834243 TI - Incidence and spectrum of chromosome abnormalities in spontaneous abortions: new insights from a 12-year study. AB - PURPOSE: Despite advances in harvesting and culturing techniques, analysis of the impact of these improvements on the observed frequency of chromosomal abnormalities in spontaneous abortions (SAB) has not been determined. We sought to evaluate the effect of these refinements on the success rate of our cultures and on the resulting frequency of detected chromosomal abnormalities. METHODS: Between 1990 and 2002, 2301 specimens obtained from the products of conception (POC) of SABs were submitted to our laboratory for cytogenetic analysis. Due to refinements in specimen processing and culture techniques introduced at the end of 1997, our data were analyzed for two periods: Period A from 1990 through 1997 with 907 eligible specimens and Period B from 1998 through 2002 with 1273 eligible specimens. RESULTS: Modifications in physician communication and sample processing contributed to significant improvements in the culture success rate and in the ratio of male-to-female cases with normal karyotypes. Additionally, increased detection of trisomic, triploid, and multiple aneuploid cases in Period B resulted in a significant increase in the percentage of cases with abnormal karyotypes (42.8% in Period A vs. 65.8% in Period B). Monosomy X accounted for < 10% of all abnormalities in Period B. Eighty five multiple aneuploid karyotypes, including 57 double trisomies, comprised 7.7% of our 1099 abnormal cases. These karyotypes were detected predominantly in POCs from the older women in our study. This collection of multiple aneuploidies is the largest published to date and includes abnormalities not reported in prior studies. We also present a table empirically derived from the data in Period B that indicates the likelihood of a specific abnormal karyotype based on maternal age. The table can be utilized by health care providers, who counsel patients after a spontaneous miscarriage. CONCLUSION: Improvements in laboratory technique have led to reduced contamination and growth failure of POCs, irrespective of maternal age. This in turn has led to a more balanced male-to-female ratio and to the detection of an increased number of abnormal cases. PMID- 15834244 TI - Detection and calibration of microdeletions and microduplications by array-based comparative genomic hybridization and its applicability to clinical genetic testing. AB - PURPOSE: Genome-wide telomere screening by fluorescence in situ hybridization (FISH) has revealed that approximately 6% of unexplained mental retardation is due to submicroscopic telomere imbalances. However, the use of FISH for telomere screening is labor intensive and time consuming, given that 41 telomeres are interrogated. We have evaluated the use of array-based Comparative Genomic Hybridization (aCGH) as a more efficient tool for identifying telomere rearrangements. METHODS: In this study, 102 individuals with unexplained mental retardation, with either normal or abnormal FISH results, were selected for a blinded retrospective study using aCGH. Results between the two methodologies were compared to ascertain the ability of aCGH to be used in a clinical diagnostics setting. RESULTS: We detected 100% of all imbalances previously identified by FISH (n = 17) and identified two additional abnormalities, a 10q telomere duplication and an interstitial duplication of 22q11. Interphase FISH analysis verified all abnormal array results. We also demonstrated that aCGH can accurately calibrate the size of telomere imbalances by using an array with "molecular rulers" for the telomeric regions of 1p, 16p, 17p, and 22q. CONCLUSION: This study demonstrates that aCGH is an equivalent methodology to telomere FISH for detecting submicroscopic deletions. In addition, small duplications that are not easily visible by FISH can be accurately detected using aCGH. Because aCGH allows simultaneous interrogation of hundreds to thousands of DNA probes and is more amenable to automation, it offers an efficient and high throughput alternative for detecting and calibrating unbalanced rearrangements, both of the telomere region, as well as other genomic locations. PMID- 15834245 TI - MMP-9 microsatellite polymorphism and susceptibility to exudative form of age related macular degeneration. AB - PURPOSE: To assess if a polymorphism (PM) of the microsatellite (CA(13-27)) in the promoter region of Matrix Metalloproteinase 9 (MMP-9) was associated with the exudative form of age-related macular degeneration (AMD) and to its risk factors. METHODS: In 107 patients with AMD (AMD Group) and 223 age- and gender-matched controls (Control Group) with cataract, demographic, clinical data, and MMP-9 PM have been compared. RESULTS: The comparison of allelic frequencies showed a different pattern of CA repeats between AMD and Control Group (P < 0.00005), in particular the prevalence of longer microsatellites (> or = 22 CA repeats) was higher in AMD than in Control Group (O.R. 2.49, 95% CI 1.71-3.37, P < 0.001). Analyses of genetic frequencies gave similar results. Logistic regression confirmed that 22 or more CA repeats are associated to AMD. The only association between MMP-9 PM and other risk factors for AMD was with BMI (Spearman's R = 0.298, P < 0.00005): all patients with both microsatellites > or = 22 CA repeats were overweight or obese (chi2 test P < 0.0005, compared to other genotypes). CONCLUSIONS: Longer microsatellites in the promoter of MMP-9 are associated to the exudative form of AMD and to body mass index, a well-known risk factor for the disease. PMID- 15834246 TI - Detection of 677CT/1298AC "double variant" chromosomes: implications for interpretation of MTHFR genotyping results. AB - PURPOSE: Genotyping 37,026 individuals as part of a thrombophilia evaluation, we determined and analyzed the genotypic frequencies of the 677CT and 1298AC mutations in the methylenetetrahydrofolate reductase (MTHFR) gene. METHODS: The 677CT and 1298AC mutations in the MTHFR gene were determined by either a laboratory-developed test involving PCR amplification and restriction digestion utilizing the ABI 3100 capillary electrophoresis apparatus (Applied Biosystems Inc) or by using an Analyte Specific Reagent (ASR) supplied by Third Wave Technologies. The genotype for three specimens with triple variant MTHFR mutations were confirmed by DNA sequencing on the ABI 3100 capillary electrophoresis apparatus. RESULTS: The MTHFR frequencies of the 677CT/1298AA, 677CC/1298AC, 677CT/1298AC, 677CC/1298AA, 677TT/1298AA, 677CC/1298CC, 677TT/1298AC, and 677CT/1298CC genotypes were 0.228, 0.208, 0.198, 0.153, 0.122, 0.088, 0.0005, and 0.0003, respectively. CONCLUSIONS: Individuals containing double variant MTHFR mutations on one allele (cis) cannot be distinguished between compound heterozygotes (trans) for 677CT and 1298AC mutations in routine clinical testing, a genotype associated with thrombophilia. Such patients could be inappropriately counseled for being at high risk for thrombotic episodes. Until information regarding prevalence and the clinical consequences of this double variant (cis) allele becomes available, caution should be used in interpreting the genotyping results of compound heterozygosity for 677CT and 1298AC. PMID- 15834247 TI - Folic acid and neural tube defects. PMID- 15834252 TI - Running a successful campaign against unionization. PMID- 15834253 TI - End-of-life care. PMID- 15834254 TI - Public reporting, patient safety, and quality improvement: the need for legal protections. PMID- 15834255 TI - Leveraging nurse-related dashboard benchmarks to expedite performance improvement and document excellence. AB - Using nursing quality benchmarks in operational dashboards and translating those data to drive performance excellence is a strategic imperative. Since access to unit-level, hospital-generated nurse-related benchmarks is an emerging arena, the authors provide an overview of aggregated trends and benchmarks gleaned from the California Nursing Outcome Coalition acute care database for 2 established nurse related quality indicators-patient falls incidence and hospital-acquired pressure ulcer prevalence. Integrating these acute care benchmarks into clinical dashboards can be invaluable to clinicians, administrators, and policy makers who share a common commitment to expediting evidence-based improvement in patient care safety, outcomes, and excellence. PMID- 15834256 TI - What really matters to healthcare consumers. AB - Consumer satisfaction with healthcare is an important quality and outcome indicator. Satisfaction may be at the crux of survival for healthcare delivery systems because it creates the competitive edge in healthcare. To better understand patient satisfaction by examining consumer healthcare experiences and expectations, a study was conducted. An important concept identified in the data, MY CARE, refers to a constellation of quality healthcare features that were wished for by all participants and realized by only some of them. The features of MY CARE offer lessons for all healthcare leaders to use when making improvements in care delivery systems-improvements that could create a more patient-centered healthcare system and boost patient satisfaction. PMID- 15834257 TI - Enhancing the quality of supportive supervisory behavior in long-term care facilities. AB - The practices of managers and registered nurses (RNs) in long-term care facilities are frequently ineffective in assisting the licensed practical nurses (LPNs) and healthcare aides (HCAs) whom they supervise. Little research exists that examines the area of supportive relationships between nursing staff and supervisors in these settings. The purpose of this study was to gather data that could improve management practices in long-term care residential facilities and enhance the quality of the supervisory relationships between supervisors (nurse managers and RNs) and care providers (HCAs and LPNs) in these settings. The study also identified factors that influence the supervisors' ability to establish supportive relationships with care providers. The challenges and barriers to nurse managers and leaders related to enacting supportive behaviors are discussed as well as their implications for long-term care settings. PMID- 15834258 TI - Revising the Essentials of Magnetism tool: there is more to adequate staffing than numbers. AB - In the Essentials of Magnetism (EOM) study, staff nurses in 9 of 16 hospitals known to have excellent staffing structures, ie Magnet hospitals, perceived staffing on their units as less than adequate. These hospitals did not fit the Magnet profile when compared to 26 Magnet, Magnet-aspiring, and non-Magnet hospitals. In the EOM study, the process "perception of adequacy of staffing" (PAS) was measured with a single-item indicator. A multi-item scale incorporating the results of a delivery system survey reported here, as well as other factors known to affect PAS, was constructed and evaluated. Results indicate that the 6 item PAS Scale is valid and reliable and is a more accurate measure of PAS than is a single-item indicator. The Magnet hospitals scored significantly higher on all 6 items of the scale than did the comparison hospitals. The 2 Magnet hospitals that did not score in the Magnet profile in the EOM study scored similarly to the Magnet hospital that scored very high in the Magnet profile. Measurement of the PAS process indicates whether staffing structures are viable and enabling and may account for the mixed results seen in some studies assessing the impact of staffing structures on patient outcomes. Discussion includes suggestions on how to promote smooth handoffs and accountability with flex delivery models, and emphasizes the need to evaluate the relationship between staff nurses' perceptions of adequate staffing and patient outcomes. PMID- 15834259 TI - Savings sharing: rewarding staff for responsible decision-making. AB - Shortages of professional nurses create a "buyer's market" in which nurses accept temporary assignments for the highest rates and offer little additional time to the primary employer. Use of temporary personnel use salary dollars at an inordinate rate while offering little continuity or support for the organization's standards. Methods for placing decision-making in the hands of the nurses are needed along with a reward system for establishing a pattern of sound decision-making. The author describes a savings sharing program that is gaining credibility in one organization for addressing both objectives. PMID- 15834260 TI - The impact of protocol on nurses' role stress: a longitudinal perspective. AB - OBJECTIVE: The study examined the impact of a protocol directed at increasing organ donation on the role stress and work attitudes of critical care nurses involved in potential organ donation cases. The research examined whether the protocol could positively affect nurses' perceptions of role stress, and if so, could the work environment improvements be sustained over time. BACKGROUND: The Family Communication Coordinator (FCC) protocol promotes effective communication during potential organ donation cases using a multidisciplinary team approach. Previous research found it associated with improved donation outcomes and with improved perceptions of role stress by critical care nurses. However, the previous study lacked methodological rigor necessary to determine causality and sustainability over time. METHODS: The study used a quasi-experimental prospective longitudinal design. The sample included critical care nurses who had experience with potential organ donation cases with the protocol. Survey data were collected at 4 points over 2 years. Surveys used previously validated and reliable measures of role stress (role ambiguity, role conflict, role overload) and work attitudes (commitment, satisfaction). Interviews supplemented these data. RESULTS: The nurses' perceptions of role stress associated with potential organ donation cases dramatically dropped after the protocol was implemented. All measures of role stress, particularly role ambiguity and role conflict, showed statistically significant and sustained improvement. Nurses' professional, unit, and hospital commitment and satisfaction reflect an increasingly positive workplace. CONCLUSIONS: The results demonstrate that the FCC protocol positively influenced the workplace through its impact on role stress over the first 2 years following its implementation. The findings suggest that similar protocols may be appropriate in improving the critical care environment by reducing the stress and uncertainty of professionals involved in other end-of-life situations. However, the most striking implication relates to the reality of the workplace: meeting the goals of improved patient care outcomes and those of improving the healthcare work environment are not mutually exclusive and may be mutually essential. PMID- 15834261 TI - Introduction to the impact of co-occurring disorders and violence on women: findings from the SAMHSA Women, Co-occurring Disorders and Violence Study. PMID- 15834262 TI - The assessment of trauma history in women with co-occurring substance abuse and mental disorders and a history of interpersonal violence. AB - The Women, Co-occurring Disorders, and Violence Study (WCDVS) was a large (N = 2729) multisite study of the effectiveness of integrated and trauma-informed services for women with substance use and mental health disorders and a history of interpersonal violence (physical or sexual abuse). Study participants' exposure to lifetime and current traumatic events was assessed at baseline and follow-up via in-person interviews. This article describes the choice of the Life Stressor Checklist-Revised (LSC-R) to assess trauma history to meet the WCDVS's research aims and to respond to consumer input. Quantitative data address the breadth and prevalence of potentially traumatic events in the past and current lives of study participants, the formation and properties of summary measures, and test-retest reliability. Qualitative data address tolerance of the instrument by interviewers and respondents and the generalizability of quantitative findings about trauma prevalence. Finally, recommendations are offered for improvements to the WCDVS version of the LSC-R for use in future research. PMID- 15834264 TI - Building resilience in children of mothers who have co-occurring disorders and histories of violence: intervention model and implementation issues. AB - Historically, children of parents with co-occurring substance abuse and mental health disorders and histories of violence/trauma have been overlooked in behavioral health treatment systems. The Women, Co-occurring Disorders and Violence Study (WCDVS) was a 5-year initiative funded by the United States Substance Abuse and Mental Health Services Administration (SAMHSA) that included a Children's Study that explored the treatment needs of children of women with these multiple disorders. This article describes the development of the Children's Study intervention that included clinical assessment, group intervention, and resource coordination/advocacy for children aged 5-10 to build resilience through increasing coping skills, improving interpersonal relationships, and helping coalesce positive identity and self-esteem. Innovative procedures, including the participation of consumer/survivor/recovering women and mothers, in the planning, implementation, and administrative applications of this intervention and study are also highlighted. It is recommended that programs begin to implement family-focused integrated treatment approaches that can potentially increase protective factors for children affected by parental mental illness, substance abuse, and violence. PMID- 15834263 TI - Physical health burdens of women with trauma histories and co-occurring substance abuse and mental disorders. AB - This article documents the physical health burdens of participants in a large, federally funded cross-site study of specialized services for women with histories of trauma (physical or sexual abuse) and co-occurring substance abuse and mental health disorders. Nearly half of the 2729 women in the study (48%) reported serious physical illnesses that frequently limited their daily life activities or required them to use special equipment. Nearly half (46%) rated their health status as only fair or poor. Given the prevalence of physical illnesses in this population, behavioral service providers should discuss with clients their overall health and how it might hinder their participation in treatment for trauma, substance abuse, and mental illness, and policymakers should consider this need when designing behavioral health requirements, setting reimbursement rates, and allocating funds. PMID- 15834265 TI - The relationship between mothers' child abuse potential and current mental health symptoms: implications for screening and referral. AB - This analysis examined data from mothers at 2 of the 9 sites participating in Substance Abuse and Mental Health Services Administration's (SAMHSA's) national Women Co-occurring Disorders and Violence Study (WCDVS). According to previous literature, it was hypothesized that women in the WCDVS would be at high risk of perpetrating child abuse. This research examined mothers' potential for physical child abuse and assessed the association between child abuse potential, current mental health symptoms, alcohol and drug use severity, and trauma. Results revealed that participants had significant potential for child abuse. Hierarchical regression analyses revealed that current mental health symptoms were the strongest predictor of mothers' scores on the Child Abuse Potential (CAP) Inventory. This study highlights the important relationships between commonly used instruments across the mental health, substance, and child welfare fields and the potential dual use of these instruments. Implications for policy and practice are discussed. PMID- 15834266 TI - The role of coercion in the treatment of women with co-occurring disorders and histories of abuse. AB - Debate continues on issues of involuntary treatment for individuals with behavioral healthcare problems. Women with co-occurring disorders and histories of abuse are an especially vulnerable population. This study seeks to increase our knowledge about the experiences of coercion for women in the behavioral healthcare system. Patterns of coercion are explored. This study did not find the predicted relationship between high levels of interpersonal violence and frequent involuntary treatment experiences. The results do offer support for the hypothesis that women are more likely to be currently mandated to treatment if they have been recently arrested, and that being mandated to treatment does not appear to be related to clinical issues such as recidivism and acute symptoms. As expected, women currently required to be in treatment report having less choice in other aspects of their care. Implications for future research in the current climate of increasingly coercive policies are presented. PMID- 15834267 TI - Social roles in women's lives: changing conceptions of self. AB - As part of an effort to improve services and outcomes for women with histories of trauma and co-occurring mental health and substance abuse disorders, the Franklin County Women and Violence Project sought to assess women's perceptions about their social roles and provide them with opportunities to adopt valued social roles. Social role theory suggests that individuals with devalued roles may benefit through the acquisition of valued roles. This article outlines the findings regarding the women's initial assessments of their social roles as well as real and desired changes 3 months after the baseline interview. Findings suggest that valued social roles play a significant role in helping women heal and recover, contributing to greater self-esteem, confidence, and happiness. Conversely, the adoption of devalued roles or worsening of roles contributes to depression and makes it more difficult for women to heal and recover. PMID- 15834269 TI - Religious/spiritual coping among women trauma survivors with mental health and substance use disorders. AB - This study examines the types of religious/spiritual coping used by women trauma survivors with co-occurring mental health and substance use disorders. Analyses based on data from 2 large racially diverse samples indicate that women from the study population rely considerably more on positive, than negative, religious coping, and that their reliance on religious coping, in general, is significantly higher than that of the general population. Numerous significant relationships were also found between the severity of trauma-related and mental health symptoms and more negative religious coping. This study further suggests that more frequent childhood abuse and childhood sexual violence are especially associated with negative religious coping in adulthood. Findings support the importance of spiritual coping for women trauma survivors with co-occurring disorders and suggest the value of increased attention to spirituality in behavioral health services, especially in assessment and therapeutic relationships. PMID- 15834268 TI - Tangled in a web of affiliation: social support networks of dually diagnosed women who are trauma survivors. AB - The goal of this article is to describe the social support networks of women with co-occurring substance abuse and mental health problems who are survivors of interpersonal abuse, using baseline interview data from 2 sites (n = 644) from the national Women Co-occurring Disorders and Violence Study. The size and composition of women's networks, the tangible and socioemotional support available, and the stance of the support network toward substance use, treatment, and trauma are described. Family members are described by women as offering less emotional support and less encouragement for healing from trauma than friends. Analyses demonstrated only modest support in either sample for the hypothesis that support network characteristics moderate the effects of traumatic stress on mental health and trauma symptoms among these samples of very burdened and poor women. The results point to the need for using caution in relying on women's existing social support network to help them heal. PMID- 15834270 TI - Relational systems change: implementing a model of change in integrating services for women with substance abuse and mental health disorders and histories of trauma. AB - This article describes the "relational systems change" model developed by the Institute for Health and Recovery, and the implementation of the model in Massachusetts from 1998-2002 to facilitate systems change to support the delivery of integrated and trauma-informed services for women with co-occurring substance abuse and mental health disorders and histories of violence and empirical evidence of resulting systems changes. The federally funded Women Embracing Life and Living (WELL) Project utilized relational strategies to facilitate systems change within and across 3 systems levels: local treatment providers, community (or region), and state. The WELL Project demonstrates that a highly collaborative, inclusive, and facilitated change process can effect services integration within agencies (intra-agency), strengthen integration within a regional network of agencies (interagency), and foster state support for services integration. PMID- 15834271 TI - Optimizing therapeutic strategies to achieve renal and cardiovascular risk reduction in diabetic patients with angiotensin receptor blockers. AB - The major challenge for the treatment of hypertensive patients with type 2 diabetes is to achieve the uniformly recommended blood pressure goal of 130/80 mmHg, and 120/75 mmHg in proteinuric patients. Such low target blood pressure levels require the administration of multiple drugs. Angiotensin receptor blockers and the combination of angiotensin receptor blockers with diuretics fulfil the criteria to lower blood pressure effectively with a placebo-like side effect profile. Beyond pressure control, clinical prospective trials have documented that it does matter what kind of antihypertensive agent is used to control blood pressure. Large-scale follow-up trials have documented blood pressure independent effects of angiotensin receptor blocker on cardiac [left ventricular hypertrophy (LVH), congestive heart failure] and renal protection (proteinuria, chronic renal failure). Of note, in these trials, angiotensin receptor blockers have been combined with diuretics, and most of the included patients have been on combination therapy comprising two to four antihypertensive agents. In addition to the combination of an angiotensin receptor blocker with a diuretic, the combination of an angiotensin receptor blocker with an angiotensin converting enzyme inhibitor appeared to be most effective in reducing proteinuria, attenuating chronic renal failure and treating congestive heart failure. PMID- 15834272 TI - Does nocturnal blood pressure fall in people of African and South Asian descent differ from that in European white populations? A systematic review and meta analysis. AB - OBJECTIVE: To assess whether nocturnal blood pressure fall in people of African (Black) and South Asian descent differs from that of the European origin white populations (White). METHODS: A systematic literature review was carried out using Medline 1966-2003 and Embase 1980-2003, and citations from references. The meta-analysis was performed using Cochrane review manager software (RevMan version 4.2; The Cochrane Collaboration, Oxford, UK). RESULTS: Seventeen studies were identified; 11 studies from the USA, one from the USA and Canada, and six studies from the United Kingdom. The mean percentage systolic blood pressure (SBP) nocturnal fall was below 10% (non-dipping) in 10 of 17 studies (59%) and the diastolic blood pressure (DBP) nocturnal fall was below 10% in four of 16 studies (25%) in Blacks compared with four of 17 studies (24%) in SBP and none in DBP nocturnal falls in Whites. Compared with Whites, Blacks had a significantly lower mean percentage nocturnal fall; the overall weighted mean difference in SBP was -3.07 (95% confidence interval, -3.81, -2.33; P < 0.00001) and in DBP was 2.98 (95% confidence interval, -3.97, -2.00; P < 0.00001). Two studies on South Asians showed a higher SBP but a similar mean DBP nocturnal fall compared with Whites. CONCLUSION: Smaller nocturnal blood pressure falls and a higher prevalence of non-dipping may contribute to the higher levels of hypertension complications seen in Black people. No such phenomenon was seen in South Asians but more research is needed to explore their higher stroke mortality. PMID- 15834273 TI - Blood pressure response to chronic intake of coffee and caffeine: a meta-analysis of randomized controlled trials. AB - PURPOSE: Coffee is a widely consumed beverage and small health effects of substances in coffee may have large public health consequences. It has been suggested that caffeine in coffee increases the risk of hypertension. We performed a meta-analysis of randomized controlled trials of coffee or caffeine and blood pressure (BP). DATA IDENTIFICATION: BP trials of coffee or caffeine published between January 1966 and January 2003 were identified through literature databases and manual search. STUDY SELECTION: A total of 16 studies with a randomized, controlled design and at least 7 days of intervention was selected, comprising 25 strata and 1010 subjects. DATA EXTRACTION: Two persons independently obtained data on sample size, type and duration of intervention, changes in BP and heart rate (HR), and subjects' characteristics for each trial. Meta-analysis was performed using a random-effects model. RESULTS: A significant rise of 2.04 mmHg [95% confidence interval (CI), 1.10-2.99] in systolic BP and 0.73 mmHg (95% CI, 0.14-1.31) in diastolic BP was found after pooling of coffee and caffeine trials. When coffee trials (n = 18, median intake: 725 ml/day) and caffeine trials (n = 7, median dose: 410 mg/day) were analysed separately, BP elevations appeared to be larger for caffeine [systolic: 4.16 mmHg (2.13-6.20); diastolic: 2.41 mmHg (0.98-3.84)] than for coffee [systolic: 1.22 mmHg (0.52 1.92) and diastolic: 0.49 mmHg (-0.06-1.04)]. Effects on HR were negligible. CONCLUSIONS: Regular caffeine intake increases BP. When ingested through coffee, however, the blood pressure effect of caffeine is small. PMID- 15834274 TI - Studies on genes and hypertension: a daunting task. PMID- 15834275 TI - Arginase is an enzyme that nephrologists can love. PMID- 15834276 TI - Angiotensin II as the link between nitric oxide and neuroadrenergic function. PMID- 15834277 TI - To D or not to D: calcitriol and vascular calcification in end-stage renal disease. PMID- 15834278 TI - Determinants of hypertension-related cardiac organ damage: beyond the haemodynamic overload. PMID- 15834279 TI - Angiotensinogen and angiotensin-converting enzyme gene copy number and angiotensin and bradykinin peptide levels in mice. AB - OBJECTIVE: To test the hypothesis that changes in gene expression that may accompany angiotensinogen (AGT) and angiotensin-converting enzyme (ACE) gene polymorphism cause alteration in angiotensin and bradykinin peptide levels. DESIGN: Mice with one or two genes for AGT and ACE allow assessment of the effects of modest alteration in AGT and ACE gene expression on angiotensin and bradykinin peptide levels. METHODS: Angiotensin and bradykinin peptides were measured in the blood, kidney, heart, lung, adrenal, brain, and aorta of mice that were either wild-type (+/+), heterozygous (+/-) or null (-/-) for either the AGT or ACE gene. RESULTS: Angiotensin I and angiotensin II were not detectable in blood or tissues of AGT -/- mice, which had increased bradykinin levels in kidney and lung. ACE -/- mice had markedly reduced angiotensin II levels and increased bradykinin levels in blood and tissues. However, despite reduced AGT and ACE gene expression, angiotensin and bradykinin peptide levels in AGT and ACE +/- mice were no different from the levels in wild-type mice. CONCLUSION: Although the AGT and ACE genes are fundamental determinants of angiotensin and bradykinin peptide levels, compensatory mechanisms attenuate the effect of modest change in AGT and ACE gene expression on the levels of these peptides. Identification of these compensatory mechanisms may provide new candidate genes for investigation in humans. PMID- 15834280 TI - Hypertension susceptibility genes on chromosome 2p24-p25 in a general Japanese population. AB - BACKGROUND: Genome-wide scans from Italy and China suggest a hypertension susceptible locus between D2S2278 (nucleotides 11,245,080 - 11,245,358) and D2S168 (nucleotides 11,467,214 - 11,467,422) on chromosome 2. METHODS: We performed a large association study of polymorphisms in this region with blood pressure modulation in a Japanese general population. Forty-seven polymorphisms in 14 genes between nucleotide 8,845,292 and nucleotide 11,946,689, which contains D2S2278 and D2S168, were genotyped in 1880 individuals, 796 of whom were hypertensive and 1084 normotensive. RESULTS: Multivariate logistic regression analysis with adjustment for age, body mass index, presence of hyperlipidemia, diabetes mellitus, and current smoking and drinking revealed that one single nucleotide polymorphism (SNP), IMS-JST126186, in HPCAL1 (hippocalcin-like 1) in women and two SNPs, IMS-JST149391 and IMS-JST149390, in GREB1 (gene regulated by estrogen in breast cancer 1) in men were significantly associated with both prevalence of hypertension and blood pressure levels. To examine the role of GREB1 in more detail, we identified 38 additional genetic variations in GREB1 by direct sequencing, and eight polymorphisms were genotyped. One SNP, 45718A>G, was significantly associated with hypertension and blood pressure level in men, and this SNP was in linkage disequilibrium with a SNP present at the 3' splice site of intron 11. CONCLUSION: Our study suggests that GREB1 and HPCAL1 are candidate hypertension-susceptibility genes in the Japanese general population and supports previous studies that also identified hypertension-related loci in this narrow region. PMID- 15834281 TI - Epistatic interaction between alpha- and gamma-adducin influences peripheral and central pulse pressures in white Europeans. AB - BACKGROUND: Adducin is a membrane skeleton protein consisting of alpha- and beta- or alpha- and gamma-subunits. Mutations in alpha- and beta-adducin are associated with hypertension. In the European Project on Genes in Hypertension, we investigated whether polymorphisms in the genes encoding alpha-adducin (Gly460Trp), beta-adducin (C1797T) and gamma-adducin (A386G), alone or in combination, affected pulse pressure (PP), an index of vascular stiffness. METHODS: We measured peripheral and central PP by conventional sphygmomanometry and applanation tonometry, respectively. We randomly recruited 642 subjects (162 nuclear families and 70 unrelated individuals) from three European populations. In multivariate analyses, we used generalized estimating equations and the quantitative transmission disequilibrium test. RESULTS: Peripheral and central PP averaged 46.1 and 32.6 mmHg, respectively. Among carriers of the alpha-adducin Trp allele, peripheral and central PP were 5.8 and 4.7 mmHg higher in gamma adducin GG homozygotes than in their AA counterparts, due to an increase in systolic pressure. gamma-Adducin GG homozygosity was associated with lower urinary Na/K ratio among alpha-adducin Trp allele carriers and with higher urinary aldosterone excretion among alpha-adducin GlyGly homozygotes. Sensitivity analyses in founders and offspring separately, and tests based on the transmission of the gamma-adducin G allele across families, confirmed the interaction between the alpha- and gamma-adducin genes. CONCLUSIONS: In alpha adducin Trp allele carriers, peripheral and central PP increased with the gamma adducin G allele. This epistatic interaction is physiologically consistent with the heterodimeric structure of the protein and its influence on transmembranous sodium transport. PMID- 15834282 TI - Arginase inhibition reduces endothelial dysfunction and blood pressure rising in spontaneously hypertensive rats. AB - OBJECTIVE: A decrease in nitric oxide (NO) bioavailability has been proposed to contribute to endothelial dysfunction and increased peripheral resistances during essential arterial hypertension. Given that arginine is a substrate for both arginase and NO synthase, arginase activity may be a critical factor in NO bioavailability. To test this hypothesis, we evaluated the effects of the arginase inhibitor alpha-difluoromethylornithine (DFMO) in spontaneously hypertensive rats (SHR). METHODS: Vascular reactivity experiments were performed on thoracic aortic rings from 10-week-old SHR and their normotensive counterparts, Wistar-Kyoto (WKY) rats. Blood pressure was measured by the tail cuff method. DFMO treatment (30 mg/kg daily in drinking water) was started in 5 week-old SHR and maintained for 5 weeks. Aortic arginase I and arginase II expression as well as arginase activity were evaluated by western blotting and the spectrophotometric method, respectively. RESULTS: DFMO (1.2 x 10 mol/l) enhanced the vascular response to acetylcholine both in SHR (+24%, P < 0.01) and WKY rats (+12%, P < 0.01), and reversed the effects of the NO synthase inhibitor N-nitro-L-arginine-methyl-ester. The vasorelaxant response to sodium nitroprusside on endothelium-denuded rings was not affected by DFMO, neither in SHR nor in WKY rats. In SHR, DFMO prevented the increase in blood pressure and improved the response of aortic rings to acetylcholine. Finally, as compared with WKY rats, SHR exhibited increased expression of vascular arginase I (+72%, P < 0.05) and arginase II (+91%, P < 0.05) as well as increased arginase activity (+26%, P < 0.05). CONCLUSIONS: Our results showed that arginase inhibition reduced endothelial dysfunction and blood pressure rising in SHR. PMID- 15834283 TI - Endothelin B receptor-deficient mice develop endothelial dysfunction independently of salt loading. AB - BACKGROUND: Rodents without a functional endothelin B (ETB) receptor develop salt sensitive hypertension. The underlying mechanisms, however, are so far unknown. The ETB receptor is involved in endothelial function by modulating the activity of the endothelial nitric oxide synthesis as well as contributing to the control of endothelial prostacyclin synthesis. In the present study, we analysed whether salt alters endothelial function in rescued ETB receptor-deficient mice. We used mice with a rescue of the lethal phenotype of an ETB knockout. These mice were generated by crossbreeding ETB mice with dopamine-hydroxylase ETB transgenic mice. METHODS: Adult rescued ETB-deficient mice were kept in parallel with wild type control animals for 15 days on standard (0.2% NaCl) or salt-enriched (4% NaCl) chow, respectively. Systolic blood pressure was measured by the tail cuff method and endothelium-dependent and endothelium-independent vascular function was assessed in isolated aortic rings under isometric conditions. RESULTS: Systolic blood pressure increased on salt-enriched chow in ETB receptor-deficient mice (166 +/- 12 mmHg), but neither in wild-type mice on high-salt diet (128 +/- 11 mmHg; P < 0.05) nor in ETB receptor-deficient mice on standard chow. The heart rate was similar in all groups at any point of time. Endothelium-dependent relaxation was impaired in ETB receptor-deficient mice (74 +/- 3 versus 96 +/- 5% of preconstriction for wild-type mice; P < 0.05) and was not significantly affected by a salt-enriched diet. Endothelium-independent relaxation was similar among all groups. Contractions to endothelin-1 were not significantly influenced by preincubation with the ETB receptor antagonist BQ-788, but were completely blunted by preincubation with the ETA receptor antagonist BQ-123 in all animals. CONCLUSION: Rescued ETB receptor-deficient mice develop salt-sensitive hypertension. Nevertheless, in this animal model of ETB receptor deficiency, endothelial function is impaired independent of salt-enriched diet or hypertension. This indicates that, in this model, salt-induced hypertension is not mediated by endothelial dysfunction. PMID- 15834284 TI - Role of endothelin in noradrenaline-induced hypertension in rats. AB - OBJECTIVE: To investigate the role of endothelin in noradrenaline-induced hypertension in rats. DESIGN: The dose-response relationship of chronic noradrenaline infusion on arterial pressure was characterized to identify a dose that would produce sustained hypertension, and the effect of combined endothelin ETA and ETB receptor blockade (TAK-044) on the response to this dose was then examined. METHODS AND RESULTS: Noradrenaline (or vehicle) was infused intravenously at 1 (subpressor acutely), 24 or 48 microg/kg per h (acute pressor response of 9 +/- 1 and 11 +/- 1 mmHg, respectively) for a 14-day infusion, and blood pressure was measured by radiotelemetry. Noradrenaline infusion at 1 microg/kg per h did not produce a 'slow pressor' rise in blood pressure. During noradrenaline infusions at 24 and 48 microg/kg per h, mean arterial pressure peaked initially on days 2-3 (+10 +/- 1 and 14 +/- 2 mmHg, respectively; P < 0.01), fell towards basal levels after day 3, and then began to rise again at days 5-6 only with 48 microg/kg per h, being 10 +/- 1 mmHg above control levels at days 13-14 (P < 0.05). TAK-044 treatment did not alter the magnitude of the initial (13 +/- 1 mmHg) or eventual (12 +/- 2 mmHg) rise in blood pressure achieved in response to 14 days' infusion of noradrenaline at 48 microg/kg per h, but abolished the transient fall. CONCLUSION: Chronic noradrenaline infusion at acutely pressor doses leads either to a transient blood pressure elevation at a moderate dose, or to a triphasic but sustained hypertension at a higher dose, with a temporary escape from the hypertension apparently mediated by endothelin. PMID- 15834285 TI - Enhanced levels of platelet P-selectin and circulating cytokines in young patients with mild arterial hypertension. AB - BACKGROUND: Emerging evidence links inflammation to atherosclerosis (AS). Although some studies have addressed the role of inflammation in patients with arterial hypertension (AH), its overall contribution in AH is far from being understood. Therefore, the present pilot study was designed to examine the role of platelet P-selectin and various inflammatory mediators in young patients with moderate AH without signs of target organ damage. METHODS AND RESULTS: Fifteen patients with mild AH [33.8 +/- 7.3 years, mean arterial pressure (MAP) 106.6 +/- 10.4 mmHg] and 15 healthy normotensive controls (31.7 +/- 10.6 years) were examined. Platelet P-selectin was analysed by flow cytometry. Plasma levels of monocyte-chemoattractant-protein-1 (MCP-1), high-sensitivity C-reactive protein (hsCRP), interleukin (IL)-6, tumour necrosis factor alpha (TNFalpha), and IL-10 levels were measured by enzyme immunoassay (EIA). Patients with mild AH showed significantly enhanced expression of platelet P-selectin [17.2 +/- 5.4 versus 10.6 +/- 4.2 mean fluorescence intensity (MFI), P < 0.001]. P-selectin expression positively correlated with MAP (r = 0.43, P < 0.05). Furthermore, patients with mild AH had significantly enhanced plasma levels of hsCRP (2.7 +/- 3.8 versus 0.6 +/- 0.9 mg/l, P < 0.01), IL-6 (1.4 +/- 0.7 versus 0.6 +/- 0.3 pg/ml, P < 0.001), TNFalpha (2.8 +/- 0.7 versus 2.4 +/- 0.4 pg/ml, P < 0.05), and MCP-1 (291.3 +/- 100.7 versus 214.3 +/- 8.3 pg/ml, P < 0.05). IL-6 levels positively correlated with hsCRP levels (r = 0.47, P < 0.05) and mean arterial pressure (MAP) (r = 0.44, P < 0.05). CONCLUSIONS: This pilot study demonstrates that in an early stage of AH, inflammatory pathways are already activated. Besides pro inflammatory cytokines, platelets seem to play a significant role in mediating inflammation in AH, which could lead to target organ injury. Further investigations have to clarify the role of early anti-inflammatory therapy, in patients with mild to moderate AH, in alleviating hypertensive target organ damage. PMID- 15834286 TI - Stress-induced increases in interleukin-6 and fibrinogen predict ambulatory blood pressure at 3-year follow-up. AB - BACKGROUND: The biological mechanisms underlying the association between psychological stress and hypertension are poorly understood. Increased plasma concentrations of the inflammatory proteins interleukin-6 and fibrinogen are commonly reported both in hypertensive patients and in people subject to chronic psychological stress. Recent laboratory studies have also shown that acute psychological stress increases plasma interleukin-6 and fibrinogen concentrations in healthy individuals. OBJECTIVE: To investigate the relationship between stress induced inflammatory responses and blood pressure using a longitudinal design. METHODS: Participants were 153 individuals from the Whitehall II cohort. Blood pressure, plasma interleukin-6 and fibrinogen were assessed in response to an acute laboratory stressor, and ambulatory blood pressure was monitored on a separate day. Three years later, a follow-up day of ambulatory blood pressure monitoring was carried out. RESULTS: Individual differences in systolic pressure, fibrinogen and interleukin-6 stress responses predicted ambulatory blood pressure at the 3-year follow-up. Larger increases in ambulatory systolic pressure over the 3-year period were predicted by larger acute fibrinogen and interleukin-6 stress responses, independently of previous ambulatory blood pressure, acute blood pressure stress responses, age, sex, body mass and smoking. CONCLUSION: Given the important roles of interleukin-6 and fibrinogen in hypertensive pathophysiology, these results indicate that psychological stress could promote hypertension through stimulating these inflammatory proteins. PMID- 15834287 TI - Role of nitric oxide in modulating systemic pressor responses to different vasoconstrictors in man. AB - OBJECTIVE: Animal studies suggest that nitric oxide (NO) attenuates responses to endogenous vasoconstrictors. We investigated whether this also holds true in man by monitoring pressor responses to different vasoconstrictors during nitric oxide synthase (NOS) inhibition. METHODS: Systemic hemodynamic responses to intravenous infusions of three doses (each for 5 min) of angiotensin II (AngII) (2, 4 and 8 ng/kg per min), noradrenaline (NOR) (10, 30 and 70 ng/kg per min) and phenylephrine (PE) (0.5, 1.0 and 1.5 microg/kg per min) were monitored in 44 healthy subjects during saline. A second dose-response curve was obtained during NOS inhibition with a subpressor dose of N- nitro-L-arginine-methyl ester (L NAME) (5 microg/kg per min) or during a systemic NO clamp using combined systemic infusions of L-NAME (12.5 microg/kg per min) and nitroprusside. Blood pressure was measured in the brachial artery and other hemodynamic parameters were derived from this signal. RESULTS: Mean arterial pressure (MAP) increased 2 +/- 2, 6 +/- 1 and 16 +/- 2 mmHg in response to AngII during saline, 7 +/- 6, 15 +/- 5 and 26 +/- 6 mmHg during the subpressor dose of L-NAME (P < 0.05) and 11 +/- 10, 18 +/- 7 and 25 +/- 6 mmHg during the systemic NO clamp (P < 0.001). These augmented responses of MAP were due to enhanced increments in systemic vascular resistance. Infusions of NOR and PE during saline resulted in dose-dependent increments in MAP and systemic vascular resistance. These increments were of comparable magnitude as those seen during AngII, but were not affected by NOS inhibition. CONCLUSION: Our findings show that the systemic pressor response evoked by AngII, but not by NOR or PE, is enhanced during NOS inhibition, suggesting that AngII is associated with increased NO release that counteracts its blood pressure rising effect. PMID- 15834288 TI - Potentiation of L-NAME-induced systemic and renal vasoconstrictor responses by alpha1-adrenoceptor antagonism. AB - BACKGROUND: Acute inhibition of nitric oxide synthase results in systemic and renal vasoconstriction, which might be attributable to unopposed activity of the sympathetic nervous (SNS) and the renin-angiotensin system (RAS). We studied the effects of N-nitro-L-arginine methyl ester (L-NAME) during alpha1-adrenoceptor blockade and concomitant angiotensin II type 1 (AT1)-receptor blockade in hypertensive individuals pretreated with hydrochlorothiazide (Hct; 25 mg once daily). METHODS: Thirteen individuals (47 +/- 9 years) were studied during administration of placebo, and after pretreatment with Hct + doxazosin (Dox; 8 mg twice daily for 9 days), with Hct + Dox + losartan (Los; 50 mg twice daily for 9 days), or (n = 5) with doxazosin or Dox + Los without hydrochlorothiazide. Mean arterial pressure (MAP) and cardiac output were derived from the finger blood pressure signal recorded by Finapres. Systemic vascular resistance (SVR) was calculated as MAP/cardiac output. Five renal clearance studies of 40 min were performed. Renal vascular resistance (RVR) was calculated as MAP divided by renal blood flow (RBF). L-NAME (12.5 microg/kg per min intravenously) was given during the third clearance period. RESULTS: MAP was 113 +/- 11 mmHg at baseline and decreased to 99 +/- 10 mmHg during the administration of Hct + Dox and to 92 +/- 10 mmHg during Hct + Dox + Los. This decrease in MAP was caused by a decrease in SVR (P = 0.0009). Pretreatment with Hct + Dox or Hct + Dox + Los had no effect on glomerular filtration rate or RBF. Infusion of L-NAME during the administration of Hct + Dox resulted in an augmented (P < 0.0001) increase in MAP (18%), SVR (61%) and RVR (70%) compared with those observed with placebo (8, 30 and 49%, respectively). This augmentation was abolished by losartan. CONCLUSION: L-NAME induced systemic and renal vasoconstrictor responses are potentiated during alpha1-adrenoceptor blockade. This potentiation was abolished by AT1-receptor antagonism. In man, unopposed activity of the SNS or SNS and RAS is not involved in the systemic and renal vasoconstriction induced by L-NAME. PMID- 15834289 TI - Cyclo-oxygenase-2, endothelium and aortic reactivity during deoxycorticosterone acetate salt-induced hypertension. AB - OBJECTIVE: To test the hypothesis that the enhanced vascular responsiveness to norepinephrine that occurs during deoxycorticosterone acetate (DOCA)-salt induced hypertension is causally related to increased expression of cyclo-oxygenase (COX) 2 and oxidative stress, which diminishes the vasomodulatory influence of endothelium-derived nitric oxide. METHODS: Four groups of age-matched, male Sprague-Dawley rats were studied: Sham (normotensive); DOCA-salt (hypertensive); DOCA-salt treated with manganese(III) tetra(4-benzoic acid) porphyrin chloride [MnTBAP, an antioxidant; 15 mg/kg intraperitoneally (i.p.) for 21 days]; DOCA salt treated with {N-[2-(cyclohexyloxy)-4-nitrophenyl]-methane sulfonamide} (NS 398, a COX-2 selective blocker; 5 mg/kg i.p. for 7 days). Contraction and relaxation were measured with FT03 force transducers coupled to a Grass polygraph in aortic rings bathed with physiologic salt solution (37 degrees C) and bubbled with a 5%CO2/95%O2 gas mixture. Aortic sensitivities (pD2 values) to norepinephrine and serum isoprostanes (8-iso-prostaglandin F2alpha, a marker of oxidative stress) were measured for each experimental paradigm. RESULTS: NS-398 significantly reduced maximal contractions in response to norepinephrine in aortic rings from Sham (44 +/- 3%) and DOCA-salt (96 +/- 2%) group rats. Expression of COX-2 protein increased significantly in vessels from DOCA-salt rats compared with those from Sham group rats. Treatment of DOCA-salt rats with either MnTBAP or NS-398 alleviated hypertension, normalized aortic pD2 values for norepinephrine and restored serum 8-isoprostane concentrations towards those observed in Sham group rats. CONCLUSIONS: COX-2 expression increases during DOCA salt hypertension, and mediates production of factors that enhance rat aortic contractility in response to norepinephrine. Our data also suggest a role for increased oxidative stress, which is at least in part dependent on enhanced COX-2 expression, in the mechanism(s) of enhanced aortic contractility in response to norepinephrine during DOCA-salt hypertension. PMID- 15834291 TI - Quality control of B-mode ultrasonic measurement of carotid artery intima-media thickness: the European Lacidipine Study on Atherosclerosis. AB - BACKGROUND: The European Lacidipine Study of Atherosclerosis (ELSA) was a prospective, randomized, double-blind, multinational interventional trial using B mode ultrasound measurement of carotid intima-media thickness (IMT) in 2259 hypertensive individuals. ELSA showed that 4-year treatment with the calcium antagonist, lacidipine, significantly slowed down progression of carotid atherosclerosis as compared with treatment with the beta-blocker, atenolol. OBJECTIVE: To report data on cross-sectional and longitudinal quality control of the ultrasound measurements implemented throughout ELSA. METHODS: Patients underwent scans at baseline and at each annual visit. All endpoints were measured while the study was in progress (initial measurements). In addition to the cross sectional quality control procedures, a longitudinal quality control procedure of re-reading 250 baseline scans at yearly intervals was implemented, to control possible reader drift. After the study had been completed, the primary endpoint was measured again under the same condition of cross-sectional quality control. RESULTS: Cross-sectional quality control data showed high reliability for all endpoints at all time points except for single maximum IMT (Tmax) and internal carotid IMT. Within-reader reliability was constantly better than between-reader reliability but, for the primary endpoint, between-reader reliability remained excellent. Initial and longitudinal quality control measurements showed a time trend toward lower IMT values. After application of a correction factor calculated from longitudinal quality control, all initial measurements no longer decreased with time, and the corrected IMT measurements were reasonably similar to those made after completion of the study. CONCLUSION: For long-term epidemiological studies and clinical trials, both cross-sectional and longitudinal quality control are critical to the reliability of measurements. In order to evaluate the absolute change in IMT and interpret study results without bias, both must be implemented. PMID- 15834290 TI - Negative regulation of RhoA/Rho kinase by angiotensin II type 2 receptor in vascular smooth muscle cells: role in angiotensin II-induced vasodilation in stroke-prone spontaneously hypertensive rats. AB - OBJECTIVE: To test whether angiotensin II (Ang II) through the Ang II type 2 receptor (AT2R), downregulates RhoA/Rho kinase, which plays a role in AT1 receptor (AT1R)-mediated function. METHODS: In vitro studies were performed in A10 vascular smooth muscle cells (VSMC) and in vivo studies in mesenteric arteries from Wistar-Kyoto (WKY) and stroke-prone spontaneously hypertensive (SHRSP) rats. VSMC were stimulated with Ang II (10 mol/l), CGP42112A (10 mol/l, a selective AT2R agonist) +/- valsartan (10 mol/l, an AT1R antagonist), or the Rho kinase inhibitor fasudil (10 mol/l). AT1R and AT2R expression and myosin light chain (MLC) phosphorylation were determined by immunoblotting. RhoA activity was assessed by measuring membrane translocation. Functional significance between AT2R, RhoA/Rho kinase and vasodilation was assessed in arteries from valsartan treated (30 mg/kg per day, 14 days) WKY and SHRSP rats. Vasodilatory responses to Ang II (10-10 mol/l) were performed in norepinephrine pre-contracted vessels +/- valsartan(10 mol/l), PD123319 (10 mol/l, an AT2R antagonist) or fasudil (10 mol/l). RESULTS: A10 VSMC expressed AT1R and AT2R. In valsartan-treated cells, Ang II-induced RhoA translocation was reduced versus controls (42 +/- 6%, P < 0.05). Similar responses were obtained with CGP42112A (45 +/- 6%, P < 0.05). This was associated with decreased MLC activation. Fasudil abrogated Ang II- and CGP42112A-mediated effects. Ang II evoked a significant vasodilatory response only in valsartan-treated SHRSP (max dilation 40 +/- 7%). PD123319 blocked these effects. Fasudil increased AngII-induced relaxation in SHRSP vessels. AT2R expression was increased by valsartan (two- to three-fold) in SHRSP arteries. RhoA translocation was increased two-fold in untreated SHRSP (P < 0.05) and was reduced by valsartan (P < 0.05). These changes were associated with decreased MLC phosphorylation. CONCLUSIONS: Ang II/AT2R negatively regulates vascular RhoA/Rho kinase/MLC phosphorylation. These processes may play a role in Ang II-mediated vasodilation in conditions associated with vascular AT2R upregulation, such as in SHRSP chronically treated with AT1R blockers, which may contribute to blood pressure lowering by these antihypertensive agents. PMID- 15834292 TI - Nocturnal blood pressure decrease is associated with increased regional cerebral blood flow in patients with a history of ischemic stroke. AB - BACKGROUND: It remains controversial whether there is a J-shaped relationship between blood pressure and recurrent stroke among patients with a recent history of ischemic stroke. OBJECTIVE: To investigate the relationship between regional cerebral blood flow (rCBF) and the dipping of nocturnal blood pressure in patients receiving antihypertensive treatment after ischemic stroke. METHODS AND RESULTS: Forty-seven patients with a previous history of ischemic stroke and 37 patients with a history of non-ischemic stroke underwent 24-h ambulatory blood pressure monitoring and rCBF measurement with single photon emission computed tomography. Of the 47 patients with ischemic stroke, 30 were diagnosed as having suffered atheromatous or embolic stroke, and 37 had an ischemic lesion in the territory of the carotid artery. Systolic and diastolic blood pressures during daytime and night-time were controlled at less than 140/90 mmHg by a low-salt diet or long-acting antihypertensive agents, or both. In patients with ischemic stroke, there were significant negative correlations between the percentage change in nocturnal blood pressure and rCBFs in the thalamus (r = -0.33, P = 0.02), putamen (r = -0.34, P = 0.02) and cerebral cortex (r = -0.31, P = 0.03). Multivariate analysis revealed that only the percentage change in nocturnal blood pressure was related to rCBF. There was a significant positive correlation between rCBFs in the thalamus and the cerebral cortex (r = 0.74, P < 0.05). In patients with non-ischemic stroke, there was no significant correlation between the percentage change in nocturnal blood pressure and rCBFs. CONCLUSIONS: These findings indicate that the decrease in nocturnal blood pressure is associated with the increase in rCBF in patients with a history of ischemic stroke in the territory of the carotid artery. PMID- 15834293 TI - Comparative effects of the angiotensin II receptor blocker, telmisartan, and the angiotensin-converting enzyme inhibitor, ramipril, on cerebrovascular structure in spontaneously hypertensive rats. AB - OBJECTIVE: Antihypertensive treatment with angiotensin-converting enzyme inhibitors (ACEIs) reverses cerebral arteriolar remodeling, thus restoring dilatation and hence the lower limit of cerebral blood flow (CBF) autoregulation (LLCBF). The objective of this study was to determine whether angiotensin II receptor AT1 blockers (ARBs) produce the same effect. DESIGN: We examined the effects of treatment with an ARB [telmisartan (TEL), 1.93 +/- 0.04 mg/kg per day] or an ACEI [ramipril (RAM), 1.00 +/- 0.02 mg/kg per day] on the cerebral circulation in spontaneously hypertensive rats (SHR). METHODS: Arteriolar pressure and diameter (cranial window) and CBF (laser Doppler) were measured during stepwise hypotensive hemorrhage, before and after deactivation (ethylenediamine tetraacetic acid), in untreated Wistar-Kyoto (WKY) rats and SHR untreated or treated for 3 months with TEL or RAM in the drinking water. RESULTS: Treatment normalized arteriolar internal diameter (SHR, 38 +/- 3 microm; TEL, 52 +/- 2 microm; RAM, 50 +/- 2 microm; WKY, 58 +/- 4 microm), essentially by reversing eutrophic inward remodeling, and the LLCBF (SHR, 80 +/- 11 mmHg; TEL, 60 +/- 4 mmHg; RAM, 71 +/- 6 mmHg; WKY, 57 +/- 5 mmHg). CONCLUSION: The fact that the ARB (TEL) is as effective as an ACEI (RAM) in reversing cerebral arteriolar remodeling suggests that the cerebrovascular AT1 receptor is an underlying mechanism that promotes hypertensive eutrophic inward remodeling. PMID- 15834294 TI - Systemic cardiovascular disease in uremic rats induced by 1,25(OH)2D3. AB - OBJECTIVE: Vitamin D may contribute to cardiovascular disease in the absence of hypercalcemia in patients with chronic kidney disease. METHODS: We investigated the effects of long-term (6-week) treatment with 1,25(OH)2D3, at a non hypercalcemic dosage (0.25 microg/kg per day per orally) in 5/6 nephrectomized rats: (i) vehicle-treated, sham-operated rats; (ii) 1,25(OH)2D3-treated, sham operated rats; (iii) vehicle-treated, 5/6 nephrectomized rats; and (iv) 1,25(OH)2D3-treated, 5/6 nephrectomized rats. RESULTS: Creatinine clearance after 6 weeks was significantly lower and parathyroid hormone levels were significantly higher in 1,25(OH)2D3-treated uremic rats, compared with uremic controls (P < 0.01). Serum calcium levels, as well as the calcium-phosphorus product, did not differ between both groups. Mean systolic blood pressure in 1,25(OH)2D3-treated animals was significantly increased, compared with vehicle (each P < 0.01). In addition, 1,25(OH)2D3-treated uremic animals showed left ventricular hypertrophy. Diffuse aortic calcification involving the intima and media layer occurred in 1,25(OH)2D3-treated uremic animals, but not in other groups. The mean aortic wall area and lumen area were increased two-fold in 1,25(OH)2D3-treated uremic animals compared with vehicle (P < 0.01), whereas the wall/lumen ratio remained unchanged, indicating fusiform aneurysm formation. CONCLUSIONS: Hypertension, left ventricular hypertrophy, aortic calcification, and aneurysm, without hypercalcemia, occurred in 1,25(OH)2D3-treated, 5/6 nephrectomized rats. These data indicate a permissive effect of uremia for cardiovascular damage induced by non-hypercalcemic doses of 1,25(OH)2D3. PMID- 15834295 TI - Angiotensin II and norepinephrine release: interaction and effects on the heart. AB - BACKGROUND: Angiotensin (Ang) II may enhance the influence of the sympathetic nervous system at various levels by facilitating norepinephrine (NE) release. We investigated whether such an interaction is evident in the human heart and whether it has an impact on left ventricular (LV) structure. METHODS AND RESULTS: Ang I and Ang II concentrations were determined in arterial and coronary sinus (CS) plasma samples in a group of normotensive (n = 10) and hypertensive (n = 18) subjects. Total systemic and cardiac NE spillover was measured using isotope dilution methodology and LV structure by echocardiography. Arterial and CS concentrations of Ang I and Ang II were similar in both groups (Ang II CS, 5.8 +/ 4.0 versus 3.7 +/- 3.1 fmol/ml; P = not significant), as was the Ang II/Ang I ratio (CS, 0.56 +/- 0.17 versus 0.54 +/- 0.22 fmol/fmol; P = not significant). Total systemic (223 +/- 145 versus 374 +/- 149 ng/min; P < 0.05) and cardiac NE spillover (11.7 +/- 6.3 versus 19.4 +/- 10.5 ng/min; P < 0.05) were increased in hypertensive patients, as was LV mass index (LVMI) (86.7 +/- 14.7 versus 117.2 +/ 19.4 g/m; P < 0.001). LVMI correlated with cardiac NE spillover (r = 0.47; P < 0.02). No correlation was evident between CS Ang II and cardiac NE spillover (r = 0.001; P = not significant) or LVMI (r = -0.20; P = not significant). Arterial Ang II tended to correlate with total systemic NE spillover (r = 0.34; P = 0.081). When hypertensive subjects were divided into two groups with either high or low CS Ang II concentration, cardiac NE spillover and LVMI did not differ between the two groups. CONCLUSION: These findings suggest a growth-promoting effect of increased cardiac sympathetic tone on cardiomyocytes in hypertensive patients, but do not support the notion of a significant role of Ang II for norepinephrine release and LV hypertrophy in the hypertensive human heart. PMID- 15834296 TI - Opposite effects of losartan and atenolol on natriuretic peptides in patients with hypertension and left ventricular hypertrophy: a LIFE substudy. AB - BACKGROUND: Secretion of natriuretic peptides is related to cardiac wall stress and influenced by the renin-angiotensin system. Therefore, we investigated the influence of blood pressure (BP) reduction with losartan versus atenolol on N terminal pro-atrial natriuretic peptide (Nt-proANP) and N-terminal pro-brain natriuretic peptide (Nt-proBNP). METHODS: In 183 patients with hypertension and electrocardiographic left ventricular (LV) hypertrophy, enrolled in the LIFE Study, we measured BP and serum Nt-proANP and Nt-proBNP by immunoassay after 2 weeks of placebo treatment and after 1, 2, 4, 6, 12, 24, 36 and 48 months of randomized treatment with losartan- or atenolol-based antihypertensive regimens. RESULTS: There was no significant difference in BP at any time point between the two treatment groups. In patients treated with losartan, median Nt-proANP decreased gradually throughout the study, reaching significance after 6 months of treatment (1125-1060 pmol/l, P < 0.001), and Nt-proBNP decreased within the first month (24.7-18.7 pmol/l, P < 0.01) and stayed reduced throughout the study. During losartan-based antihypertensive treatment, Nt-proANP and Nt-proBNP as a percentage of baseline values were correlated to reductions in systolic BP (r = 0.11, P < 0.01 and r = 0.10, P = 0.01) and diastolic BP (r = 0.17, P < 0.001 and r = 0.07, P = 0.09). In atenolol-treated patients, Nt-proANP (1100-1640 pmol/l, P < 0.001) and Nt-proBNP (20.0-37.7 pmol/l, P < 0.001) increased during the first month, and remained elevated throughout the study. During atenolol-based antihypertensive treatment, changes in Nt-proANP (r = -0.16, P < 0.001) and Nt proBNP (r = -0.07, P = 0.08) were negatively related to change in heart rate. CONCLUSION: Nt-proANP and Nt-proBNP were reduced in parallel with BP in losartan treated patients whereas they increased in parallel with decreased heart rate in atenolol-treated patients. PMID- 15834297 TI - Cardiac structural and functional changes during long-term antihypertensive treatment with lacidipine and atenolol in the European Lacidipine Study on Atherosclerosis (ELSA). AB - OBJECTIVES: To evaluate and correlate the effects of long-term antihypertensive treatment on left ventricular (LV) mass and carotid structural changes in a large group of essential hypertensive patients, participating in the European Lacidipine Study on Atherosclerosis (ELSA). DESIGN: In four (Brescia, Glasgow, Naples and Pisa) of 23 centres participating in the ELSA study, an echocardiographic examination was performed at baseline and repeated, until the end of the 4-year study, in essential hypertensive patients, followed-up for carotid quantitative ultrasound examination of intima-media thickness (IMT), after random allocation to treatment with either lacidipine or atenolol (and added hydrochlorothiazide, as required for control of blood pressure). METHODS: M mode, two-dimensional guided echocardiography was used to measure left ventricular (LV) wall thickness and dimensions, from which LV mass was calculated, using an anatomically validated formula (Penn Convention) and indexed to body surface area (left ventricular mass index, LVMI). The echocardiographic tracings were blindly evaluated in a single reading centre (Brescia). Bilateral IMT was measured at the site of common carotid and bifurcation far walls (CBMmax). RESULTS: At baseline, cardiac and carotid ultrasound scans were available in 278 patients (mean age 54 +/- 7 years, 57% males, 22% obese). A significant correlation was observed between baseline LVMI and CBMmax (r = 0.22, P < 0.001), independent of age. In multivariate analysis, CBMmax and mean 24-h pulse pressure were most strongly associated with baseline LVMI. A significant reduction in LVMI was observed both during lacidipine (n = 96) (-12.5% reduction) and atenolol (n = 78) (-13.9% reduction) treatments (up to 4 years) (P < 0.001 for both, without significant differences between treatments). Changes in LVMI were not related to changes in carotid wall thickness. In multivariate analysis, baseline LV mass and mean 24-h systolic blood pressure changes were significantly associated with changes in LV mass. CONCLUSIONS: In this large, long-term controlled study, antihypertensive treatment with atenolol or lacidipine was accompanied by a similar and significant decrease in LV mass. Treatment-induced changes in LV mass were related to baseline LV mass and changes in 24-h mean systolic blood pressure, without any correlation with changes in carotid structure. In the whole ELSA population, carotid IMT changes have been shown to be unrelated to blood pressure reduction, but significantly influenced by the type of antihypertensive treatment. PMID- 15834298 TI - Results of the Controlled ONset Verapamil INvestigation of Cardiovascular Endpoints (CONVINCE) trial by geographical region. AB - OBJECTIVE: To examine regional differences in the Controlled ONset Verapamil INvestigation of Cardiovascular Endpoints (CONVINCE) trial. DESIGN: Double-blind, randomized, international clinical trial. SETTING: Six hundred and sixty-one clinical centers in 15 countries. PATIENTS: Hypertensive volunteers (n = 16,602) with > or =1 additional cardiovascular risk factor, grouped into four regions: USA (n = 8144), Canada (n = 3405), Western Europe (Spain, UK, Italy, Sweden, Germany; n = 2048) or 'other' (Bulgaria, Israel, Mexico, Czech Republic, Hungary, Poland, Slovakia, Brazil; n = 2879); subgroupings included country and state/province within the USA and Canada. INTERVENTIONS: Randomized to COER verapamil or the investigator's choice of either atenolol or hydrochlorothiazide, titrated and additional drugs added as required. MAIN OUTCOME MEASURES: Baseline characteristics; blood pressure control, medication adherence and lost-to-follow up at 2 years; and composite primary endpoint (stroke, myocardial infarction, cardiovascular death) by regional groupings. RESULTS: Regional differences were found at baseline for age, gender, blood pressure, percentage receiving antihypertensive drug therapy, initial choice of atenolol or hydrochlorothiazide, and risk factor profile. Blood pressure control rates increased markedly during follow-up in all regions, but varied significantly by region. Blood pressure control, medication adherence and lost-to-follow-up rates were poorest in the USA. After adjustment for baseline differences, the primary-event rate for each region was significantly lower than for the USA. Although baseline factors, blood pressure control and event rates varied by region, treatment differences did not. CONCLUSION: Despite differences in baseline and follow-up measures across geographical regions, the absence of treatment differences by region suggests that the overall findings of CONVINCE are robust. PMID- 15834299 TI - Reply to critics of research on transcendental meditation in the prevention and control of hypertension. PMID- 15834301 TI - Facts and fiction of poor compliance as a cause of inadequate blood pressure control. PMID- 15834303 TI - Peroxisome proliferator-activated receptor-gamma is a new therapeutic target in sepsis and inflammation. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the nuclear receptor superfamily and a ligand-activated transcription factor with pleiotropic effects on lipid metabolism, inflammation, and cell proliferation. PPARgamma forms a heterodimer with the retinoid X receptor and upon ligand activation binds to the PPAR response element in the promoter of genes to allow transcription. The class of insulin-sensitizing drugs known as thiazolidinediones have been identified as specific PPARgamma agonists that have allowed the characterization of many genes regulated by PPARgamma. Thiazolidinediones include rosiglitazone, pioglitazone, troglitazone, and ciglitazone. In addition to these synthetic agonists, cyclopentenone prostaglandins of the J2 series have been identified as natural ligands for PPARgamma. Several in vitro and in vivo studies have demonstrated that pharmacological activation of PPARgamma by 15-deoxy Delta(12,14)-PGJ2 (15d-PGJ2) or thiazolidinediones has anti-inflammatory effects. This article provides an overview of the role of PPARgamma in regulating the inflammatory response and emphasizes the potential efficacy of PPARgamma ligands as novel therapeutic approaches beyond diabetes in sepsis, inflammation, and reperfusion injury. PMID- 15834304 TI - A pilot-controlled study of a polymyxin B-immobilized hemoperfusion cartridge in patients with severe sepsis secondary to intra-abdominal infection. AB - Endotoxin is an important pathogenic trigger for sepsis. The polymyxin B immobilized endotoxin removal hemoperfusion cartridge, Toraymyxin (hereafter PMX), has been shown to remove endotoxin in preclinical and open-label clinical studies. In a multicenter, open-label, pilot, randomized, controlled study conducted in the intensive care unit in six academic medical centers in Europe, 36 postsurgical patients with severe sepsis or septic shock secondary to intra abdominal infection were randomized to PMX treatment of 2 h (n = 17) or standard therapy (n = 19). PMX was well tolerated and showed no significant side effects. There were no statistically significant differences in the change in endotoxin levels from baseline to 6 to 8 h after treatment or to 24 h after treatment between the two groups. There was also no significant difference in the change in interleukin (IL)-6 levels from baseline to 6 to 8 h after treatment or to 24 h after treatment between the two groups. Patients treated with PMX demonstrated significant increases in cardiac index (CI; P = 0.012 and 0.032 at days 1 and 2, respectively), left ventricular stroke work index (LVSWI, P = 0.015 at day 2), and oxygen delivery index (DO2I, P = 0.007 at day 2) compared with the controls. The need for continuous renal replacement therapy (CRRT) after study entry was reduced in the PMX group (P = 0.043). There was no significant difference between the groups in organ dysfunction as assessed by the Sequential Organ Failure Assessment (SOFA) scores from day 0 (baseline) to day 6. Treatment using the PMX cartridge is safe and may improve cardiac and renal dysfunction due to sepsis or septic shock. Further studies are needed to prove this effectiveness. PMID- 15834305 TI - Cerebrospinal fluid concentrations of anti-inflammatory mediators in early-phase severe traumatic brain injury. AB - In our previous study of patients with early-phase severe traumatic brain injury (TBI), the anti-inflammatory interleukin (IL)-10 concentration was lower in cerebrospinal fluid (CSF) than in serum, whereas proinflammatory IL-1beta and tumor necrosis factor (TNF)-alpha concentrations were higher in CSF than in serum. To clarify the influence of additional injury on this disproportion between proinflammatory and anti-inflammatory mediators, we compared their CSF and serum concentrations in patients with severe TBI with and without additional injury. All 35 study patients (18 with and 17 without additional injury) had a Glasgow Coma Scale score of 8 or less upon admission. With the exception of additional injury, clinical characteristics did not differ significantly between groups. CSF and serum concentrations of two proinflammatory mediators (IL-1beta and TNF-alpha,) and three anti-inflammatory mediators (IL-1 receptor antagonist [IL-1ra], soluble TNF receptor-I [sTNFr-I], and IL-10) were measured and compared at 6 h after injury. CSF concentrations of proinflammatory mediators were much higher than the corresponding serum concentrations in both patient groups (P < 0.001). In contrast, serum concentrations of anti-inflammatory mediators were much higher than the paired CSF concentrations in patients with additional injury (P < 0.001), but serum concentrations were lower than or equal to the corresponding CSF concentrations in patients without additional injury. CSF concentrations of IL-1beta, IL-1ra, sTNFr-I, and IL-10 were significantly higher (P < 0.01 for all) in patients with high intracranial pressure (ICP; n = 11) than in patients with low ICP (n = 24), and were also significantly higher (P < 0.05 for all) in patients with an unfavorable outcome (n = 14) than in patients with a favorable outcome (n = 21). These findings indicate that increased serum concentrations of anti-inflammatory mediators after severe TBI are mainly due to additional extracranial injury. We conclude that anti-inflammatory mediators in CSF may be useful indicators of the severity of brain damage in terms of ICP as well as overall prognosis of patients with severe TBI. PMID- 15834306 TI - Hemorrhagic shock resuscitation affects early and selective mesenteric artery endothelial function through a free radical-dependent mechanism. AB - Mesenteric ischemia/reperfusion occurring during hemorrhagic shock and resuscitation (H/R) induces a systemic inflammatory response and damages endothelial cells. Our aim was to investigate whether H/R affects selectively mesenteric vascular reactivity and the roles of free radicals and inducible nitric oxide (NO) synthase (iNOS) in these changes. Rats subjected to H (30 min)/R (60 min) in the presence or absence of the free radical scavenger N-2 mercaptopropionyl glycine (MPG), or the specific inhibitor of iNOS [(3) N-(3 aminomethyl)benzyl) acetaminide; 1400W] were studied. Saline requirements to maintain systemic blood pressure during R (53.4 +/- 5.2 mL/kg/h) were reduced by MPG (26.2 +/- 3.1) and 1400W (37.5 +/- 4.1). H/R reduced maximal mesenteric arteries relaxation to acetylcholine (sham: 70% +/- 5%, H/R: 21% +/- 3%) and this impairment was prevented by MPG (66% +/- 10%) and reduced by 1400W (49% +/- 9%). H/R did not affect the endothelium-independent relaxations. Maximal responses to phenylephrine were reduced in mesenteric arteries by H/R (3.6 +/- 0.5 mN/mm vs. sham 6.5 +/- 0.5), this impairment was prevented by 1400W and MPG. No impaired response to acetylcholine was detected in skeletal muscle arteries. H/R was associated with an increased production of TNF-alpha (169 +/- 8.5 ng/mL vs. sham 38 +/- 5 ng/mL), and this was reduced to 75 +/- 8 ng/mL in MPG-treated rats. Total intestinal content of iNOS mRNA was also increased by H/R and this increase was partly reduced by treatment with MPG. H/R induces an early and selective mesenteric endothelial cell dysfunction through a mechanism that involves oxygen derived free radicals and NO produced by iNOS. H/R is associated with a mesenteric hyporeactivity through an induction of NOS and may be prevented by scavenging free radicals. This early impairment in endothelial function is associated with a local inflammatory response. PMID- 15834307 TI - Trauma-hemorrhagic shock mesenteric lymph from rat contains a modified form of albumin that is implicated in endothelial cell toxicity. AB - It has been proposed that factors originating from the gut after severe trauma/shock are introduced into the systemic circulation through the mesenteric lymphatics and are responsible for the cellular injury and inflammation that culminates in acute multiple organ dysfunction syndrome (MODS). Indeed, it has been shown that lymph collected from shocked but not sham-shocked animals causes endothelial cell death, neutrophil activation, and bone marrow (BM) colony growth suppression in vitro. In an attempt to isolate the factor(s) in lymph responsible for endothelial cell toxicity, lymph from shock and sham animals was fractionated by solid phase extraction (SPE) and ion exchange chromatography (IEX). The separation of shock lymph by both methodologies yielded two fractions having major detectable toxicity to endothelial cells, whereas no toxicity was detected from sham lymph separations by either method. Subsequent analysis of each SPE toxic fraction by gel electrophoresis and mass spectrometry suggests the toxicity is associated with a modified form of rat serum albumin (mod-RSA) and multiple lipid-based factors. Therefore, we have been able to demonstrate by two different separation techniques that shock lymph contains two or more factors that may account for the toxicity to endothelial cells. Further investigations are needed to determine the type of RSA modification and the identity of the lipid factors and their role in MODS. PMID- 15834308 TI - Progressive exercise preconditioning protects against circulatory shock during experimental heatstroke. AB - Heat shock protein (HSP) 72 expression protects against arterial hypotension in rat heatstroke. HSP72 can also be induced in multiple organs, including hearts from rats with endurance exercise. We validated the hypothesis that progressive exercise preconditioning may confer cardiovascular protection during heatstroke by inducing the overexpression of HSP72 in multiple organs. To deal with the matter, we assessed the effects of heatstroke on mean arterial pressure, heart rate, cardiac output, stroke volume, total peripheral vascular resistance, colonic temperature, blood gases, and serum or tissue levels of tumor necrosis factor-alpha (TNF-alpha) in urethane-anesthetized rats pretreated without or with progressive exercise training for 1, 2, or 3 weeks. In addition, HSP72 expression in multiple organs was determined in different groups of animals. Heatstroke was induced by exposing the rats to a high blanket temperature (43 degrees C); the moment at which mean arterial pressure decreased from the peak value was taken as the time of heatstroke onset. Previous exercise training for 3 weeks, but not 1 or 2 weeks, conferred significant protection against hyperthermia, arterial hypotension, decreased cardiac output, decreased stroke volume, decreased peripheral vascular resistance, and increased levels of serum or tissue TNF-alpha during heatstroke and correlated with overexpression of HSP72 in multiple organs, including heart, liver, and adrenal gland. However, 10 days after 3 weeks of progressive exercise training, when HSP72 expression in multiple organs returned to basal values, the beneficial effects exerted by 3 weeks of exercise training were no longer observed. These results strongly suggest that HSP72 preconditioning with progressive exercise training protects against hyperthermia, circulatory shock, and TNF-alpha overproduction during heatstroke. PMID- 15834309 TI - Heat shock response inhibits release of high mobility group box 1 protein induced by endotoxin in murine macrophages. AB - The purpose of this study was to evaluate the kinetic changes and the localization of high-mobility group box 1 protein (HMGB1) and to observe the effect of heat shock response (HSR) on the expression and release of HMGB1 in lipopolysaccharide (LPS)-activated murine macrophage-like RAW 264.7 cells. Reverse transcriptase (RT)-PCR and Western blot were used to examine HMGB1 expression after LPS treatment. The intracellular localization of HMGB1 in normal or LPS-activated cells was investigated by immunocytochemical analysis and HMGB1 released from cultured macrophages by Western blot. HSR was performed by incubating RAW 264.7 cells at 42.5 degrees C for 1 h then recovery at 37 degrees C for 12 h. The effect of HSR on expression and release of HMGB1 was observed. The results showed that a decrease of HMGB1 mRNA expression was observed at 18 h after LPS (500 ng/mL) treatment, although the total intracellular HMGB1 protein levels were not affected. A visible translocation of HMGB1 from the nuclear to the cytoplasm was observed at 20 h after stimulation with LPS (500 ng/mL). Furthermore, HMGB1 was released into the medium by LPS-activated RAW 264.7 cells in a time- and dose-dependent manner. Heat shock pretreatment significantly inhibited LPS-induced release of HMGB1 and the translocation of HMGB1 from the nucleus to the cytoplasm in RAW 264.7 cells. These findings suggest that the release of HMGB1 by LPS-activated macrophages is a late event in the pathogenesis of sepsis and that HSR could inhibit the release and translocation of HMGB1 induced by LPS. PMID- 15834310 TI - Dietary supplementation of soybean kunitz trypsin inhibitor reduces lipopolysaccharide-induced lethality in mouse model. AB - We examined the modifying effects of a Kunitz trypsin inhibitor (KTI) and a Bowman-Birk trypsin inhibitor (BBI), purified from soybean, as intraperitoneal (i.p.) injection and dietary supplements on bacterial lipopolysaccharide (LPS) induced lethality in mice. We initially examined the suppressing effects of i.p. injection of KTI (50 mg/kg) and BBI (50 mg/kg) on LPS-induced lethality after i.p. injection of LPS. Furthermore, groups of female C57BL/6 were fed a basal diet (control group) or the basal diet supplemented with KTI (50 g/kg) or BBI (50 g/kg). Here, we show that i.p. and daily oral administration of KTI, but not BBI, caused a significant reduction of the LPS-induced lethality; that LPS significantly induced plasma TNF-alpha, IL-1beta, and IL-6 levels in mice after LPS challenge; that concomitant administration of KTI, but not BBI, inhibits the LPS-induced plasma levels of these cytokines; and that KTI, but not BBI, suppressed LPS-induced upregulation of cytokine expression through suppression of phosphorylation of three mitogen-activated protein (MAP) kinase pathways, ERK1/2, JNK, and p38, in peritoneal macrophages. These data allow us to speculate that i.p. injection and dietary supplementation of a soybean KTI may play a role as a potent anti-inflammatory agent by inhibiting activation of MAP kinases, leading to the suppression of cytokine expression. PMID- 15834311 TI - Prolactin secretion in hypothyroid endotoxemic rats: involvement of L-arginine and nitric oxide synthase. AB - The identification of nitric oxide (NO) within the hypothalamus and pituitary gland has suggested its role as modulator of the activity on hypothalamic pituitary axis. Hypothalamic NO synthase (NOS) is known to be regulated by thyroid hormones. We investigated the effects of previous injection of N-nitro-L arginine methyl ester (L-NAME), a NOS inhibitor, and L-arginine (L-Arg), the substrate for NO synthesis, on prolactin (PRL) secretion, through the lipopolysaccharide (LPS)-induced inflammatory response in thyroidectomized (TX) rats. TX or sham-operated (N) rats were intraperitoneally (i.p.) injected with L NAME (10 mg kg) or L-Arg (200 mg kg) or the same volume of vehicle (saline solution) 30 min before endotoxemia-induction with LPS at 250 mug (100 g body weight), i.p.. In N rats, NO increased PRL release in response to endotoxemia, whereas in hypothyroid rats, NO appeared to have the opposite effect. Our data support the hypothesis that NO exerts a modulatory influence on PRL secretion after LPS-induced inflammatory response. PMID- 15834312 TI - Hepatocytes enhance effects of lipopolysaccharide on liver nonparenchymal cells through close cell interactions. AB - The response to lipopolysaccharide (LPS) in the liver is complex, requiring cell to-cell interactions between hepatocytes and liver nonparenchymal cells (NPC), in particular, Kupffer cells. Previous studies show that cytokines produced by Kupffer cells stimulated with LPS can, in turn, activate hepatocytes. In the present study, we sought to examine whether the reverse, hepatocyte (HC)-NPC interactions, is important in cytokine production in mixed cell cocultures. LPS stimulated production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 from NPC was augmented in mixed HC-NPC cocultures, as compared with NPC monocultures. This HC-NPC interaction was not observed when hepatocytes were cocultured with NPC from TLR4-mutant (C3H/HeJ) mice or CD14-deficient mice. The effect was partially lost when hepatocytes from lipopolysaccharide-binding protein (LBP)-deficient mice were cocultured with wild-type mice. These data indicate that functional TLR4 and CD14 are required for NPC production of cytokines and that at least one of the critical components from hepatocytes is LBP. The augmented cytokine production by mixed HC-NPC cocultures was abrogated when the cells were separated by a filter system, indicating that close cell interactions are also required for this interaction. Thus, interaction between hepatocytes and NPC are critical for cytokine secretion by NPC. PMID- 15834313 TI - Hypertonic saline modulates innate immunity in a model of systemic inflammation. AB - We sought to determine if hypertonic saline (HTS) impacted alveolar macrophage (AM) activation and intracellular inflammatory gene signaling in a model of systemic inflammation. Rats received an intravenous administration of 4 mL/kg of 7.5% HTS or L-lactate lactated Ringer's (L-LR). They were simultaneously treated with an intraperitoneal injection of zymosan, which induces noninfectious systemic inflammation. AM were harvested by bronchoalveolar lavage 24 h after treatment. AM activation was analyzed by measurement of baseline and lipopolysaccharide (LPS)-induced TNF-alpha production. Intracellular signaling was analyzed for activation of the mitogen-activated protein kinases (MAPKs): ERK1/2, JNK, and p38. AM from HTS-treated rats produced less TNF-alpha than from L-LR-treated rats (927 +/- 335 pg/mL [SEM] vs. 3628 +/- 783 pg/mL [SEM], P = 0.001) and were also less responsive to LPS (4444 +/- 86 pg/mL [SEM] vs. 6666 +/- 91 pg/mL [SEM], P = 0.058). However, there was no difference in MAPK activation. In vivo HTS prevents excessive AM activation during systemic inflammation. This suppression is mediated through alternate pathways and does not induce the classic MAPK signaling cascade. PMID- 15834314 TI - Early expression of heme oxygenase-1 in leukocytes correlates negatively with oxidative stress and predicts hepatic and renal dysfunction at late stage of sepsis. AB - Oxidative stress triggered by septic insult may be the major cause of multiple organ dysfunction syndrome (MODS) in intensive unit care patients. The inducible form of heme oxygenase-1 (HO-1) can be induced by cytokines, lipopolysaccharide, and reactive oxygen species during sepsis. These facts raise the question of whether the expression of HO-1 in leukocytes can indicate the level of oxidative stress of multiple organs in sepsis. Clinical peritonitis was simulated in an animal model by cecal ligation and puncture (CLP). The level of oxidative stress was examined by plasma lipid peroxidation (LPO). Liver function was analyzed by plasma aspartate aminotransferase, alanine aminotransferase, total bilirubin, and direct bilirubin. Lung function was evaluated by severity of edema. Renal function was measured by blood urea nitrogen and creatinine. The correlation between early HO-1 induction and LPO level or organ functional indicators of the same rat at late sepsis was analyzed by linear regression. The results showed that the protein content of HO-1 increased at 9 h after CLP, whereas expression of HO-1 mRNA in leukocytes was significantly increased (P < 0.01) at 6 h after CLP. Plasma level of LPO and the indices of hepatic, pulmonary, and renal function were significantly increased at 18 h after CLP. Moreover, highly negative correlations were observed between HO-1 mRNA expression at 6 h after CLP and level of LPO or severity of hepatic/renal dysfunction at 18 h after CLP. These results suggest that early HO-1 mRNA expression in leukocytes may represent oxidative stress and may predict the severity of liver and renal dysfunction during sepsis. PMID- 15834315 TI - Amelioration of the development of multiple organ dysfunction syndrome by somatostatin via suppression of intestinal mucosal mast cells. AB - Multiple organ dysfunction syndrome (MODS) is the most serious complication of trauma or infection. Our previous study has shown that activated intestinal mucosal mast cells (IMMC) might play an important role in the development of MODS. Somatostatin (SST), one of the peptides derived from gut, is an important regulator in the neuroendocrine-immune network. However, the effects of SST on IMMC, especially in the situation of MODS, remain unclear. The aim of this study was to investigate the effect of SST on the activity of IMMC in MODS. A rat model of MODS was established 24 h after intraperitoneal injection of zymosan at dosage of 75 mg/kg. SST was injected into the tail vein 30 min after intraperitoneal inoculation of zymosan. Animals were sacrificed 25 h after zymosan injection. The concentration of histamine and tumor necrosis factor-alpha (TNF-alpha) in plasma and intestinal tissue was measured. The pathological changes of vital organs, including intestine, liver, kidney, and lung, were studied under light microscopy. The ultramicrostructures of IMMC were observed by transmission electron microscopy. Obvious improvement of pathological changes of vital organs was observed in the rats with MODS treated with SST at 2.3 ng/kg/h. PO2 was increased by 50% (P < 0.05). The histamine level in the intestinal tissue of rats with MODS treated with SST (14.50 +/- 1.08 ng/g protein) was significantly higher than that of the group without treatment (8.60 +/- 0.50 ng/g protein, P < 0.01). Furthermore, degranulation of IMMC in the rats treated with SST was less obvious. The in vitro inhibitive effect of SST on the histamine release rate of IMMC was negatively correlated to its concentration (r = -0.991, P < 0.01). In conclusion, suppression of IMMC activity might be an important mechanism of the protective effects of SST in rats with a high risk of MODS. PMID- 15834316 TI - Characterization of a new rat model of experimental combined neurotrauma. AB - Traumatic brain injury (TBI) is present in two-thirds of patients with multiple injuries and in one-third combined with injuries of the extremities. Studies on interactive effects between central and peripheral injuries are scarce due to the absence of clinically relevant models. To meet the demand for "more-hit" models, an experimental model of combined neurotrauma (CNT) incorporating a standardized TBI via lateral fluid percussion (LFP) together with a peripheral bone fracture, i.e., tibia fracture, is introduced. Sprague-Dawley rats were randomized to four experimental groups: controls (n = 10), animals with TBI (n = 30), animals with tibia fracture (n = 30), and animals with CNT (n = 30). Morphological aspects of brain and bone injury were analyzed via standard histopathological procedures and x-ray. Trauma-induced neuromotor dysfunction was assessed using a standardized neuroscore. For interactive effects between injuries, we studied the extent and temporal pattern of circulating interleukin 6 (IL-6) levels via immunoassay and callus formation at fracture sites by means of microradiography. LFP produced an ipsilateral lesion with cortical contusion, hemorrhage, mass shift, and neuronal cell loss (adjacent cortex and hippocampus CA-2/-3), along with contralateral neuromotor dysfunction. X-rays confirmed complete fractures in the middle of the bone shaft. The type of injury (P < 0.001) and time (P = 0.022) were significantly associated with increased IL-6 levels. CNT produced the highest IL 6 plasma levels with a maximum peak at 6 h after trauma (P < 0.001). Similarly, callus formation at fracture sites in CNT was significantly increased versus fracture only (P < 0,01). The CNT model mimics a variety of clinically relevant features known from human multiple injury, including TBI, and offers novel approaches for investigation of interactive mechanisms and therapeutic approaches. PMID- 15834317 TI - An investigational study on the healing process of anterior spinal arthrodesis using a bioactive ceramic spacer and the change in load-sharing of spinal instrumentation. AB - STUDY DESIGN: Ceramic anterior lumbar interbody arthrodesis was performed using an in vivo sheep model. Observations of fusion status and the load-sharing of spinal instrumentation were studied at sequential intervals for 1 year after surgery. OBJECTIVES: To elucidate the healing process of spinal arthrodesis performed with a bioactive ceramic spacer and the change in load-sharing of anterior spinal instrumentation. SUMMARY OF BACKGROUND DATA: With the improved development of spinal instrumentation, anterior spinal arthrodesis has become a standard spinal reconstruction technique; however, the mechanistic basis underlying the healing process is not well documented. Moreover, it remains unclear how load-distribution through the fusion mass and spinal instrumentation change throughout the healing process. METHODS: Using 24 sheep, a two-level anterior lumbar interbody fusion (L2-L3, L4-L5) was performed using a smooth surface and a porous surface-modified bioactive ceramic, with each segment instrumented using a one-rod anterior spinal instrumentation system. Four animals each were killed at 2, 4, 8, 12, 24, and 52 weeks after surgery. Postmortem analysis included quantification of anterior rod strain under multidirectional flexibility testing and radiographic and histologic analyses of the arthrodesed segments. RESULTS: From 0 to 8 weeks after surgery, the bending strain of the rod gradually decreased despite no obvious bone formation. From 8 to 24 weeks after surgery, the rod strain markedly decreased with the development of bridging trabeculated bone formation between vertebral bodies. After 24 weeks after surgery, minimal changes were observed in rod strain; however, the fusion mass volumetrically increased with corresponding facet joint atrophy. The porous surface-modification of ceramic did not influence the histologic healing process, despite the improvement of interface osseous union rate. CONCLUSIONS: In anterior spinal arthrodesis, spinal instrumentation is mainly exposed to bending stress, with decreased load-sharing with corresponding development of the spinal fusion. Continuous bone remodeling of the anterior fusion mass results in concurrent decreases in spinal instrumentation and posterior spinal element load transmission. The principal healing mechanism of ceramic anterior interbody spinal fusion is not an osseous union between the ceramic and vertebral body, but bridging bone formation around the ceramic, which directly connects the vertebral bodies above and below the disc. PMID- 15834318 TI - Pure shear properties of lumbar spinal joints and the effect of tissue sectioning on load sharing. AB - STUDY DESIGN: An in vitro biomechanical study on lumbar intervertebral joints. OBJECTIVES: To examine the mechanical properties of lumbar motion segments under pure shear loading and establish whether a simple model for functional differentiation between the anterior column and the posterior elements is applicable. SUMMARY OF BACKGROUND DATA: Anteroposterior shear has been implicated as a major factor in spinal instability. There is a substantial amount of data on shear motion as a coupled part of flexion-extension; data on the pure shear properties of intervertebral joints is limited. METHODS: Eighteen human cadaver lumbar motion segments were subject to nondestructive testing under pure shear loads (anterior shear and posterior shear). An MTS standard testing machine was used to record the load-deformation characteristics of specimens subject to deformation at a constant rate to a maximum shear load of approximately 250 N. Tissue sectioning was then performed with the specimen mounted in the testing machine. Eight specimens were sectioned through the intervertebral disc, including the anterior and posterior longitudinal ligaments, and 8 specimens were sectioned through the pedicles to remove the posterior elements. The same deformation pattern applied to the intact specimen was then reapplied to the sectioned specimen, and the load-deformation characteristics following sectioning were evaluated. RESULTS: The shear stiffness of the intact segments were found to be higher in anterior shear (mean group A = 583.8, B = 607 N/mm) than in posterior shear (mean group A = 469, B = 438.4 N/mm). Section of the anterior column and adjacent longitudinal ligaments resulted in a mean stiffness decreased by 22.8% of the intact value under anterior shear and 23.9% under posterior shear. Much larger change in shear stiffness was seen, and the mean sectioned stiffness dropped by 77.7% in anterior shear and 79% in posterior shear after removal of the posterior elements. After the anterior column was sectioned, 12% and 18% increases in the deformation for anterior and posterior directions were seen, whereas a distinct increase in the deformations was found after posterior elements sectioned. CONCLUSIONS: The posterior elements of the lumbar spine are more efficient in resisting anterior and posterior shear loads. However, the anterior column will exhibit similar load-displacement characteristics if subject to greater deformations. The sum of the normalized mean shear loads of the anterior column and posterior elements sustained at maximum intact deformation is significantly different from the shear load sustained by the intact spine at the same deformation. A simple concept of load sharing between the anterior column and the posterior elements may not be valid. PMID- 15834319 TI - Clinical significance of congenital lumbosacral malformations in young male population with prolonged low back pain. AB - STUDY DESIGN: A prospective clinical investigation. OBJECTIVE: To determine the clinical significance of congenital lumbosacral malformations in young male patients with subacute or chronic low back pain and to investigate a possible worsening of the clinical picture with the concurrence of transitional vertebra (TV) or spina bifida occulta (SBO). SUMMARY OF BACKGROUND DATA: Although the causative roles of congenital malformations in low back pain and lumbar disc herniation have been investigated, there is no study that compares the incidence of congenital malformations in prolonged low back pain between radicular and nonradicular groups; nor is there a study that compares the pain intensity between the groups with and without congenital malformations. METHODS: Lumbosacral plain radiographs of 881 young male patients with low back pain lasting for >4 weeks were evaluated. For all patients, we recorded the clinical signs and their pain intensities based on a 10-cm visual analog scale (VAS). RESULTS: Congenital lumbosacral malformations were determined in 88 of 881 patients. Of these, 48 were TV, 38 were SBO, and 2 were a combined lesion of TV and SBO. The difference in positive clinical sign and VAS was statistically significant (P < 0.05) between the groups with and without congenital lumbosacral malformations. In contrast to SBO, there was a statistically significant difference of TV incidence between the sign positive and the sign negative groups (P < 0.05). The difference in VAS values was statistically significant for both TV and SBO (P < 0.001). CONCLUSIONS: The results of this study show that SBO and TV increase the severity of the clinical picture whether or not they have a causal relationship. Additionally, transitional vertebrae seem to increase nerve root symptoms whereas spina bifida occulta does not. PMID- 15834320 TI - Scoliosis after median sternotomy in children with congenital heart disease. AB - STUDY DESIGN: A retrospective review of spinal deformity in patients with congenital heart disease surgically treated through a median sternotomy before the age of 8 years. Assessment was done on chest roentgenograms at skeletal maturity. OBJECTIVES: To determine if patients surgically treated through a median sternotomy present a higher prevalence of spinal deformity. SUMMARY OF BACKGROUND DATA: Congenital heart disease is associated with a higher prevalence of scoliosis. The etiology of scoliosis in this group of patients is unknown. Thoracotomy causes scoliosis, but median sternotomy has not been identified as a causal agent in these patients. METHODS: Chest roentgenograms were done after skeletal maturity in 128 patients to assess for the presence of a spinal deformity in the sagittal or coronal plane as a result of a median sternotomy for treatment of congenital heart disease before the age of 8 years in patients without any prior radiographic evidence of spinal or costal deformity before surgery. RESULTS: Forty-four (34.3%) of these patients had scoliosis greater than 10 degrees , 16 of them (12.5%) had curves greater than 20 degrees , and 33 (25.8%) had thoracic kyphosis lesser than 20 degrees . Patients operated before the age of 18 months had a significantly increased risk of developing scoliosis than those operated at a later age (odds ratio = 3.5; confidence interval = 1.3 9.6; P = 0.016). The presence of scoliosis was not related to the type of congenital heart disease. CONCLUSIONS: There is a high prevalence of scoliosis in patients with congenital heart disease surgically treated through a median sternotomy. The prevalence of scoliosis increases in patients operated at an earlier age. PMID- 15834321 TI - Potential complications of posterior spine fusion and instrumentation in patients with cerebral palsy treated with intrathecal baclofen infusion. AB - STUDY DESIGN: The authors evaluated a case series of 5 patients with cerebral palsy and a previously placed subcutaneous reservoir for continuous intrathecal baclofen infusion to control spasticity who underwent spinal arthrodesis with posterior instrumentation for progressive neuromuscular scoliosis deformities. OBJECTIVES: To present our preliminary experience and complications with posterior spine fusion in patients with cerebral palsy and continuous intrathecal baclofen infusion and provide a literature review. SUMMARY OF BACKGROUND DATA: Numerous studies have reported complications associated with intrathecal baclofen infusion, many of these catheter-related. Few reports address complications associated with the intrathecal baclofen infusion, including progressive spinal deformities and complications following spinal arthrodesis. METHODS: Five patients with spastic quadriplegia cerebral palsy had spine fusions for progressive neuromuscular scoliosis. The mean age at surgery was 14.5 years. The mean preoperative major curve was 73 degrees and mean pelvic obliquity was 19 degrees . The patients' medical records were reviewed for complications following reinsertion of the intrathecal catheter following posterior spine instrumentation and arthrodesis, and for progression of the neuromuscular scoliosis, before and after intrathecal baclofen infusion was initiated. RESULTS: Two patients presented with low-pressure headaches in the postoperative period, resulting from cerebrospinal fluid leak following subarachnoid catheter reinsertion and posterior instrumentation. Four patients documented progression of the major scoliosis curve after intrathecal baclofen infusion was begun. A mean progression of 44 degrees occurred over a mean period of 11 months before the spinal arthrodesis. CONCLUSIONS: Low pressure headaches resulting from a cerebrospinal fluid leak following catheter reinsertion may occur in the postoperative period. Preoperative concerns with the baclofen pump reservoir placed subcutaneously and pressure sores were not seen with careful prone positioning on a 4-poster frame. Progression of scoliosis in patients with cerebral palsy requiring spinal arthrodesis was demonstrated in 4 of the patients after continuous intrathecal baclofen was started. The progression of the spinal deformity as a consequence of growth, natural history, or the intrathecal baclofen infusion is unknown at the present time. PMID- 15834322 TI - Adult complex spinal dysraphism with situs inversus totalis: a rare association and review. AB - STUDY DESIGN: First published report of an adult complex spinal dysraphism with situs inversus. OBJECTIVES: To describe a previously asymptomatic adult patient of multiple vertebral anomalies with cervical split cord malformation type II, tethering of the spinal cord (cervical and lumbar), and intraspinal arachnoid cyst along with dextrocardia and situs inversus. SUMMARY OF BACKGROUND DATA: Only 5 cases (fetus, 1; neonates, 3; child, 1) of spinal dysraphism with dextrocardia or situs inversus have been reported. All these cases have had associated multiorgan developmental anomalies usually incompatible with survival and requiring multidisciplinary care. METHODS: The case has been described and relevant literature reviewed. RESULTS: The patient was operated for cervical and lumbar levels in the same sitting. A C4-C5 laminectomy was performed, 2 hemicords enclosed in the same dural sac were visualized, dorsal paramedian nerve roots and the tethering arachnoid bands were cut, and the arachnoid cyst wall was partially excised. This was followed by L4-L5 laminectomy and detethering by sectioning of the thickened filum terminale. The patient showed significant neurologic improvement after surgery. CONCLUSIONS: The present case is a rare instance in which there has been an association of adult onset occult spinal dysraphism along with situs inversus totalis. Successful management requires appropriate understanding of embryology, anatomy, and imaging and has implications in neurosurgical and perioperative anesthetic care. PMID- 15834324 TI - Is there a continuing role for prospective observational studies in spine research? PMID- 15834325 TI - Effect of three different physical therapy treatments on pain and activity in pregnant women with pelvic girdle pain: a randomized clinical trial with 3, 6, and 12 months follow-up postpartum. AB - STUDY DESIGN: A randomized assessor-blinded clinical trial was conducted. OBJECTIVE: To compare 3 different physical therapy treatments with respect to pain and activity in women with pelvic girdle pain during pregnancy and 3, 6, and 12 months postpartum. SUMMARY OF BACKGROUND DATA: In spite of the high prevalence of back pain during pregnancy, documented treatment programs are limited. METHODS: Based on a clinical examination, 118 women with pelvic girdle pain diagnosed during pregnancy were randomized into 3 different treatment groups: Information Group, use of a nonelastic sacroiliac belt and oral/written information about pelvic girdle pain (n = 40); Home Exercise Group, same as in the Information Group, with the addition of a home exercise program (n = 41); and the In Clinic Exercise Group, same as in the Information Group, plus participation in a training program (n = 37). Pain intensity was rated on a visual analogue scale (0-100 mm) and marked on a pain drawing concerning localization. The activity ability was scored using the Disability Rating Index, covering 12 daily activity items. Outcome measures were obtained at inclusion, on average in gestation week 38, and 3, 6, and 12 months postpartum. RESULTS: There was no significant difference among the 3 groups during pregnancy or at the follow-ups postpartum regarding pain and activity. In all groups, pain decreased and the activity ability increased between gestation week 38 and at 12 months postpartum. CONCLUSIONS: Women with pelvic girdle pain seemed to improve with time in all 3 treatment groups. Neitherhome nor in clinic exercises had any additional value above giving a nonelastic sacroiliac belt and information. PMID- 15834326 TI - The efficacy of corticosteroids in periradicular infiltration for chronic radicular pain: a randomized, double-blind, controlled trial. AB - STUDY DESIGN: A randomized, double-blind controlled trial. OBJECTIVES: To determine the treatment effect of corticosteroids in periradicular infiltration for chronic radicular pain. We also examined prognostic factors in relation to the outcome of the procedure. SUMMARY OF BACKGROUND DATA: Various studies have examined the therapeutic value of periradicular infiltration using treatment agents consisting of local anesthetic and corticosteroids for radicular pain, secondary to lumbar disc herniation and spinal stenosis. There is currently no randomized trial to determine the efficacy of a single injection of corticosteroids for chronic radicular pain. METHODS: Eligible patients with radicular pain who had unilateral symptoms who failed conservative management were randomized for a single injection with bupivacaine and methylprednisolone or bupivacaine only. Outcome measures included the Oswestry Disability Index, visual analogue score for back pain and leg pain, claudication walking distance, and the patient's subjective level of satisfaction of the outcome. RESULTS: We recruited 43 patients in the bupivacaine and methylprednisolone group and 43 patients in the bupivacaine only group. The follow-up rate is 100%. Five patients had early termination of the trial for discectomy and further root block. There is no statistically significant difference in the outcome measures between the groups at 3 months (change of the Oswestry Disability Index [P = 0.68], change in visual analogue score [back pain, P = 0.68; leg pain, P = 0.94], change in walking distance [P = 0.7]). Duration of symptoms has a statistically significant negative association with the change in Oswestry Disability Index (P = 0.03). CONCLUSION: Clinical improvement occurred in both groups of patients. Corticosteroids did not provide additional benefit. PMID- 15834327 TI - A prospective evaluation of anesthesia for posterior lumbar spine fusion: the effectiveness of preoperative epidural anesthesia with morphine. AB - STUDY DESIGN: A prospective, randomized, single-blinded study. OBJECTIVES: To prospectively compare the anesthesia for posterior lumbar spinal fusion between the age- and gender-matched groups with and without preoperative epidural anesthesia with morphine. SUMMARY OF BACKGROUND DATA: To reduce the blood loss, hypotensive anesthesia is frequently used in the spinal fusion. However, in spite of administration of the hypotensive agents, inadequate analgesia during operation often causes unfavorable and unexpected elevation of blood pressure resulting in excessive bleeding. As well, intractable wound pain sometimes occurs after spinal fusion due to inadequate analgesia during surgery. METHODS: Forty cases of posterior lumbar spinal fusion were investigated prospectively. Twenty patients underwent preoperative epidural anesthesia with morphine and sedation by continuous intravenous administration of propofol but without administration of any kind of anesthetics, including intraoperative and/or postoperative epidural anesthesia (Group E). Another 20 patients underwent inhalation general anesthesia with sevoflurane and intermittent intravenous administration of fentanyl without any regional block (Group F). In these groups, dynamics and average mean blood pressure during surgery, blood loss during surgery, postoperative requirement of analgesics, the visual analogue scale, and accuracy of neurologic evaluation immediately after surgery were analyzed. RESULTS: The average mean blood pressure during surgery was significantly lower in Group E than Group F: 57.1 mm Hg and 72.8 mm Hg, respectively. In addition, the dynamics of blood pressure were more stable in Group E than Group F. The blood loss during surgery was significantly less in Group E than in Group F: 546 g and 631 g, respectively. The patients of Group E required analgesics after surgery less frequently, and the rating on the visual analogue scale was significantly lower in Group F on the first, second, and third days after surgery. It was difficult to evaluate the neurologic status accurately in 5 patients of Group F because of the inadequate emergence immediately after surgery. There were no patients in Group E whose neurologic status was unclear because of the paralysis caused by epidural anesthesia. CONCLUSIONS: To combine preoperative epidural anesthesia with other anesthetic procedure for spinal fusion contributes to maintain more stable hypotension during surgery. As a result, it will be possible to lessen the bleeding during surgery. In addition, the postoperative pain was easily controlled with administration of fewer analgesic agents in patients who underwent the epidural anesthesia. The effectiveness of pre-emptive analgesia should be emphasized to diminish the postoperative pain. PMID- 15834328 TI - Differential activation of the thoracic multifidus and longissimus thoracis during trunk rotation. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVE: To develop a technique to measure electromyographic (EMG) activity of deep and superficial paraspinal muscles at different thoracic levels and to investigate activity of these muscles during seated trunk rotation. SUMMARY OF BACKGROUND DATA: Few studies have compared activity of deep and superficial paraspinal muscles of the thorax during trunk rotation, and conflicting results have been presented. Conflicting data may result from recording techniques or variation in activity between thoracic regions. METHODS: EMG recordings were made from deep (multifidus/rotatores) and superficial (longissimus) paraspinal muscles at T5, T8, and T11 using selective intramuscular electrodes. Ten subjects rotated the trunk to end of range in each direction. EMG amplitude was measured in neutral, at end of range, and during four epochs, which represented four quarters of the movement. RESULTS: During trunk rotation in sitting, longissimus EMG either increased with ipsilateral rotation (T5) or decreased with contralateral rotation (T5, T8, T11). In contrast, multifidus EMG was more variable and was either active with rotation in both directions (particularly T5) or with one movement direction. CONCLUSIONS: The deep and superficial muscles of the thorax are differentially active, and the patterns of activity differ between the regions of the thorax. Data from this study support the hypothesis that multifidus may have a role in control of segmental motion at T5. Variability in multifidus activity at T8 and T11 suggests that this muscle may also control coupling between rotation and lateral flexion. PMID- 15834329 TI - The extent of ossification of posterior longitudinal ligament of the spine associated with nucleotide pyrophosphatase gene and leptin receptor gene polymorphisms. AB - STUDY DESIGN: A case-control study using radiograph findings and the PCR assay with regard to the susceptibility and the severity of ossification of posterior longitudinal ligament of the spine (OPLL). OBJECTIVE: To analyze whether polymorphisms of the nucleotide pyrophosphatase (NPPS) gene and the leptin receptor gene predispose to an increased frequency and severity of OPLL. SUMMARY OF BACKGROUND DATA: The NPPS gene is responsible for ectopic ossification in the ttw mouse, an animal model for OPLL. The Zucker fatty rat, another animal model for OPLL, has a missense mutation in the leptin receptor gene. METHODS: Analysis of 172 OPLL patients and 93 non-OPLL controls was performed. Radiographs of the cervical, thoracic and lumber spine were analyzed to determine whether OPLL was present and to what degree. Genomic DNA was extracted from all participants. Polymorphisms of the NPPS gene and the leptin receptor gene were analyzed using the PCR assay. The association of the polymorphisms with the development and extent of OPLL were statistically evaluated. RESULTS: No significant association was found between the polymorphisms and the existence of OPLL in both the NPPS and the leptin receptor genes. However, the IVS20-11delT variant in the NPPS gene and the A861G variant in the leptin receptor gene were more frequent in patients with OPLL in the thoracic spine compared with patients whose OPLL was restricted to the cervical spine. CONCLUSION: The present results suggest that the IVS20 11delT variant of the NPPS gene and the A861G variant of the leptin receptor gene are associated with more extensive OPLL, but not with the frequency with which it occurs. PMID- 15834331 TI - Nucleus pulposus cells upregulate PI3K/Akt and MEK/ERK signaling pathways under hypoxic conditions and resist apoptosis induced by serum withdrawal. AB - STUDY DESIGN: To examine the impact of hypoxia, rat nucleus pulposus cells were maintained in monolayer culture in 2% O2 and survival and signal transduction pathways identified. OBJECTIVE: To elucidate the signaling pathways that allow nucleus pulposus cells to adapt to low oxygen environment. SUMMARY OF BACKGROUND DATA: Mammalian cell function is critically dependent on a continuous supply of oxygen. Interestingly, some specialized cell types that include nucleus pulposus cells of the intervertebral disc reside in a hypoxic environment. However, the mechanism of their adaptation to this low oxygen environment is not known. METHODS: Rat nucleus pulposus cells were harvested from explant cultures and grown to confluence in monolayer. Cells from passage 3-7 were maintained under hypoxia (2% O2) and normoxia (20% O2) for various time periods in complete or serum-free medium. Cells were also treated with pharmacologic agents that block PI3K and MAPK signaling pathways. Cell survival was assessed by MTT assay, annexinV-PI dual-color flow cytometry, and the TUNEL procedure. Expression of signaling proteins was evaluated by Western blot analysis. Cell phenotype was studied by semiquantitative RT-PCR. RESULTS: Under hypoxic conditions, rat nucleus pulposus cells were resistant to apoptosis induced by serum starvation. Protection was also observed after treatment of the nucleus cells by desferrioxamine, a compound that mimics many of the effects of hypoxia. Cell survival in hypoxia was related to activation of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MEK)/extracellular signal regulated kinase (ERK) pathways. Induction of Akt activation and ERK1/2 activationunder hypoxic condition was detected at 12 hours and correlated with inactivation of glycogen synthase kinase-3beta (GSK-3beta), an effector protein involved in regulation of apoptosis. Finally, inhibition of PI3K/Akt and MEK/ERK pathway using the inhibitors LY294002 and PD98059, respectively, impaired cell survival. CONCLUSION: It is concluded that under hypoxic conditions, rat nucleus pulposus cells are adapted for survival by regulation of expression of critical genes, downregulation of apoptosis through activation of the PI3K/Akt and MAPK survival pathways. PMID- 15834332 TI - Experimental anterior lumbar interbody fusion with an osteoinductive bovine bone collagen extract. AB - STUDY DESIGN: A randomized experimental porcine anterior lumbar interbody fusion. OBJECTIVE: To determine the osteoinductive potential of a bovine bone derived collagen lyophilisate for anterior interbody spinal fusion in an animal model. SUMMARY OF BACKGROUND DATA: Complications associated with autologous bone graft harvesting are still major concerns when performing spinal fusion surgery. A new formulation of bovine bone collagen-protein extracts (Colloss) has been shown to be effective in animal studies as a bone graft enhancer. The present study further tested its osteogenic potential in the anterior spinal fusion cage in a controlled animal study. METHODS: Twelve normal Danish landrace pigs of 50 kg each were chosen as experimental animals. Lumbar spine interbody fusion of L3-L4, L4-L5 using titanium alloy cages was performed on each pig with pedicle screws instrumentation. Cages packed with either autograft or Colloss were randomly assigned to the two levels. Pigs were followed up to 3 months. After termination, the spine segments were evaluated by radiograph, micro-CT, and histomorphometry RESULTS: Ten pigs completed the study. Implant breakage, loosening, or spinal deformity was not observed after 12 weeks on radiograph examination. Micro-CT evaluations demonstrated that cages with Colloss had the same fusion rate (9 of 10) as those with autograft, further evaluation of bone volume (BV/TV), and trabecular thickness (Tb Th) also showed no different between them. Histologically, both autograft and Colloss levels had 80% (8 of 10) fusion rate. Histomorphometric results revealed that the only difference was the Colloss filled cages had a higher cartilage tissue volume (P < 0.04). CONCLUSION: Colloss was effective as a bone graft substitute. It achieved the same fusion rate and the samepercentage of new bone formation as that of autologous iliac bone graft in this demanding porcine anterior spinal fusion model after 12 weeks. PMID- 15834333 TI - Biomechanical comparison of transarticular facet screws to lateral mass plates in two-level instrumentations of the cervical spine. AB - STUDY DESIGN: In vitro biomechanical comparison of transarticular facet screws to lateral mass plates in two level instrumentations of the cervical spine. OBJECTIVE: Lateral mass plates are costly, and screw placement is difficult. Facet screws have never been tested as an alternative in the cervical spine. This biomechanical study compared cervical transarticular facet screws to lateral mass plates in two-level instrumentations of human cadaveric cervical spines. SUMMARY OF BACKGROUND DATA: Translaminar facet screws have been shown to have similar biomechanical performance to pedicle screw fixation in the lumbar spine, especially in flexion. They have proven to be fast, safe, and effective, with authors reporting 94% to 100% fusion rates in single-level lumbar fusions. However, a biomechanical comparison of transarticular facet screws to lateral mass plates in cervical spine instrumentations has not been reported. METHODS: Thirteen human cadaveric cervical motion segments (C2-C4, C5-C7) were tested before and after instrumentation, with either transarticular facet screws or lateral mass plates, in flexion, extension, lateral bending, and torsion. Specimens were subjected to six cycles under a load of 2 Nm. RESULTS: Both fixation systems significantly reduced range of motion (ROM) and increased stiffness compared with the intact state in flexion, extension, lateral bending, and torsion. There were also no significant differences between the facet screws and plates in any of the four directions. To compare the two systems, ROM of each was analyzed relative to the uninstrumented state. Flexion was 0.26 (or 26% of the intact state) for the transarticular facet screws versus 0.20 for the lateral mass plates (P = 0.34), extension was 0.10 versus 0.07 (P = 0.43), lateral bending was 0.17 versus 0.15 (P = 0.52), and torque was 0.25 versus 0.38 (P = 0.12). Load to failure testing failed to indicate any differences between the two methods of fixation because all the specimens failed elsewhere. CONCLUSION: This study proves that transarticular facet screws and lateral mass plates are equivalent in two-level instrumentations of the cervical spine. This is the first biomechanical study to test transarticular facet screws in this context. PMID- 15834334 TI - The effect of an interspinous process implant on facet loading during extension. AB - STUDY DESIGN: Facet loading parameters of lumbar cadaver spines were measured during extension before and after placement of an interspinous process implant. OBJECTIVE: The study was undertaken to quantify the influence of an interspinous implant on facet loading at the implanted and adjacent levels during extension. SUMMARY OF BACKGROUND DATA: Facet loading is increased during extension and decreased during flexion. Previous studies have demonstrated that interspinous process decompression relieves disc pressure at the implanted level and does not alter disc pressure at the adjacent levels. Facet joints are believed to play a key role in back pain, especially in patients with collapsed discs and increased motion segment mobility resulting in increased facet loading. METHODS: Seven cadaver spines (L2-L5) were loaded to 15 Nm of extension and 700 N compression with and without an interspinous process implant (X STOP) placed between the L3 L4 spinous processes. Pressure-sensitive film was placed in the facet joints of the implanted and adjacent levels. After loading, the film was digitally analyzed for peak pressure, average pressure, contact area, and force. These values were compared between the intact and implanted specimens at the adjacent and implanted levels using a paired t test (P < 0.05). RESULTS: The implant significantly reduced the mean peak pressure, average pressure, contact area, and force at the implanted level. The mean peak pressure, average pressure, contact area, and force at the adjacent levels were not significantly different between the intact and implanted specimens with the exception of contact area at the L2-L3 level. CONCLUSIONS: Interspinous process decompression will unlikely cause adjacent level facet pain or accelerated facet joint degeneration. Furthermore, pain induced from pressure originating in the facets and/or posterior anulus of the lumbar spine may be relieved by interspinous pro-cess decompression. Clinical results from patients with a component of lower back pain suggest that this is a valid conclusion. PMID- 15834335 TI - Stress shielding in box and cylinder cervical interbody fusion cage designs. AB - STUDY DESIGN: A numerical analysis of stress shielding of the bone graft in box and cylinder interbody fusion cages was performed. OBJECTIVES: To evaluate the stress shielding characteristics of box and cylinder interbody fusion cages for the cervical spine with regard to their rigidity and contiguous pore size. SUMMARY OF BACKGROUND DATA: Cage design has been shown to influence loading of the augmented bone graft tissue. In addition, a large contiguous pore design is believed to be important to avoid stress shielding effects. METHODS: A two dimensional axisymmetric, biphasic finite-element model of the cage incorporating the bone graft and the adjacent vertebral bodies was developed. Analysis was performed in two parts. First, the vertebrae were loaded by an axial compressive force, and second, the effect of vertebral penetration by the interbody cage was simulated. RESULTS: Straining of bone graft in the box cage was generally lower than that of the cylinder cage. The strains in the cylinder cage were seen to be more uniformly distributed, whereas in the box cage straining was concentrated in the graft under the endplates. Vertebral penetration by the cylinder cage resulted in significant straining of the bone graft (28% strain), whereas lower strains were determined in the box cage (a maximum of 17% strain). CONCLUSIONS: The central pore in the box design does not seem as effective as the fully open cylinder cage in transferring loads to the augmented graft tissue. Early penetration of the adjacent vertebrae by the cylinder cage may provide early postoperative stability and load the graft tissue, thereby imparting the necessary signals for fusion. PMID- 15834336 TI - Biomechanical analysis of rigid stabilization techniques for three-column injury in the lower cervical spine. AB - STUDY DESIGN: Comparison of nondestructive multidirectional flexibility in groups of specimens receiving two different posterior instrumentation constructs with or without anterior plating. OBJECTIVE: To compare stability after a three-column injury stabilized posteriorly by lateral mass screws-rods at C5-C6 and pedicle screws-rods at C7 ("LLP") or by pedicle screws-rods at C5-C6-C7 ("PPP"), and to compare posterior, anterior, and combined anterior-posterior fixation. SUMMARY OF BACKGROUND DATA: Pedicle screws resist pullout better than lateral mass screws, but little research has compared the stability of pedicle screws to that of lateral mass screws used within constructs. METHODS: Fourteen human cadaveric C4 T1 specimens were tested intact, posteriorly instrumented (7 LLP and 7 PPP), anteriorly instrumented, or with combined (anterior-posterior) instrumentation. Nonconstraining, nondestructive torques induced flexion, extension, lateral bending, and axial rotation while angular motion was recorded optically. RESULTS: Posterior, anterior, and combined instrumentation each significantly improved stability (P < 0.05). Combined fixation provided significantly better stability than either anterior or posterior instrumentation alone. In no loading mode and in no testing condition was any parameter significantly different between LLP and PPP. Posterior instrumentation provided significantly better stability than anterior instrumentation. CONCLUSIONS: Anterior plate and posterior screw-rod fixation alone improve stability in a two-level, three-column cervical injury model. Combined fixation further improves stability. There is little discernible difference in immediate postoperative stability between posterior rod constructs combining lateral mass and pedicle screws and those using only pedicle screws. PMID- 15834337 TI - Serum levels of nickel and chromium after instrumented posterior spinal arthrodesis. AB - STUDY DESIGN: Cross-sectional study of 37 patients to measure serum levels of nickel and chromium after posterior spinal arthrodesis using stainless steel implants. OBJECTIVES: To investigate the relationship between factors such as age, gender, pain, time from surgery, length of arthrodesis, and level of arthrodesis to serum metal ion levels after instrumented spinal arthrodesis. SUMMARY OF BACKGROUND DATA: Measurable levels of metal ions in the serum can be detected after the use of stainless steel implants. There is some evidence to suggest that long-term exposure can potentially be toxic. Posterior spinal arthrodesis with stainless steel implants is a common procedure to treat spinal deformity in the adolescent population; however, the extent of metal ion exposure after posterior spinal arthrodesis is unknown. METHODS: Patients that underwent posterior instrumented spinal arthrodesis with more than 6 months follow-up were recruited for this study. Patients with altered neurologic function were excluded. Serum levels of nickel and chromium were measured using inductively coupled plasma mass spectrometry. Pain was assessed using the Oswestry questionnaire. Spine radiographs were used to look for evidence of pseudarthrosis. Forty-five patients were approached, and 37 agreed to the questionnaire and blood test. Ten patients were men and 27 were women. Mean age at surgery was 14 years with mean follow-up of 6 years. Statistical correlations between serum metal ion levels and age at surgery, time from surgery, gender, number of segments fused, spinal instrument interfaces, pain, and instrumentation type were assessed. RESULTS: Abnormally high levels of nickel and chromium above normal levels (0.3 ng/mL for nickel, 0.15 ng/mL for chromium) could be detected in serum after posterior spinal arthrodesis using stainless steel implants. There was a significant inverse correlation between serum nickel (r = -0.61, P < 0.001) and chromium (r = -0.64, P < 0.001) levels and time from surgery. When patients were grouped based on lengths of time from surgery, 0 to 2 years (n = 7), 2 to 4 years (n = 11), and >4 years (n = 8), the mean +/- SD for nickel (ng/mL) was 3.8 +/- 2.6, 1.3 +/- 1.1, and 0.9 +/- 0.8, respectively. Analysis ofvariance revealed significant group differences (P =0.004). Similarly, the chromium levels were 2.7 +/- 2.7, 0.6 +/- 0.4, and 0.3 +/- 0.3, respectively (P = 0.018). Only time from surgery was a significant multivariate predictor of nickel and chromium serum levels. Pseudarthrosis was not seen in this cohort. CONCLUSIONS: Elevated levels of nickel and chromium can be measured after posterior instrumented spinal arthrodesis. The levels diminish rapidly with time from surgery but still remained above normal levels 4 years after surgery. Long-term implication of this metal ion exposure is unknown and should be studied further. PMID- 15834338 TI - Long-term outcomes of surgical and nonsurgical management of sciatica secondary to a lumbar disc herniation: 10 year results from the maine lumbar spine study. AB - STUDY DESIGN: A prospective cohort study. OBJECTIVE: To assess 10-year outcomes of patients with sciatica resulting from a lumbar disc herniation treated surgically or nonsurgically. SUMMARY OF BACKGROUND DATA: There is little information comparing long-term outcomes of surgical and conservative therapy of lumbar disc herniation in contemporary clinical practice. Prior studies suggest that these outcomes are similar. METHODS: Patients recruited from the practices of orthopedic surgeons, neurosurgeons, and occupational medicine physicians throughout Maine had baseline interviews with follow-up questionnaires mailed at regular intervals over 10 years. Clinical data were obtained at baseline from a physician questionnaire. Primary analyses were based on initial treatment received, either surgical or nonsurgical. Secondary analyses examined actual treatments received by 10 years. Outcomes included patient-reported symptoms of leg and back pain, functional status, satisfaction, and work and disability compensation status. RESULTS: Of 507 eligible consenting patients initially enrolled, 10-year outcomes were available for 400 of 477 (84%) surviving patients; 217 of 255 (85%) treated surgically, and 183 of 222 (82%) treated nonsurgically. Patients undergoing surgery had worse baseline symptoms and functional status than those initially treated nonsurgically. By 10 years, 25% of surgical patients had undergone at least one additional lumbar spine operation, and 25% of nonsurgical patients had at least one lumbar spine operation. At 10 year follow-up, 69% of patients initially treated surgically reported improvement in their predominant symptom (back or leg pain) versus 61% of those initially treated nonsurgically (P = 0.2). A larger proportion of surgical patients reported that their low back and leg pain were much better or completely gone (56% vs. 40%, P = 0.006) and were more satisfied with their current status (71% vs. 56%, P = 0.002). Treatment group differences persisted after adjustment for other determinants of outcome in multivariate models. Change in the modified Roland back-specific functional status scale favored surgical treatment, and the relative benefit persisted over the follow-up period. Despite these differences, work and disability status at 10 years were comparable among those treated surgically or nonsurgically. CONCLUSIONS: Surgically treated patients with a herniated lumbar disc had more complete relief of leg pain and improved function and satisfaction compared with nonsurgically treated patients over 10 years. Nevertheless, improvement in the patient's predominant symptom and work and disability outcomes were similar regardless of treatment received. For patients in whom elective discectomy is a treatment option, an individualized treatment plan requires patients and their physicians to integrate clinical findings with patient preferences based on their symptoms and goals. PMID- 15834340 TI - Acupuncture and dry-needling for low back pain: an updated systematic review within the framework of the cochrane collaboration. AB - OBJECTIVES: To assess the effects of acupuncture and dry-needling for the treatment of nonspecific low back pain. BACKGROUND: Low back pain is usually a self-limiting condition that tends to improve spontaneously over time. However, for many people, back pain becomes a chronic or recurrent problem for which a large variety of therapeutic interventions are employed. SEARCH STRATEGY: We updated the searches from 1996 to February 2003 in CENTRAL, MEDLINE, and EMBASE. We also searched the Chinese Cochrane Centre database of clinical trials and Japanese databases to February 2003. SELECTION CRITERIA: Randomized controlled trials of acupuncture (that involved needling) or dry-needling for adults with nonspecific acute/subacute or chronic low back pain. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed methodologic quality (using the criteria recommended by the Cochrane Back Review Group) and extracted data. The trials were combined using meta-analysis methods or levels of evidence when the data reported did not allow statistical pooling. RESULTS: Thirty-five randomized clinical trials were included: 20 were published in English, 7 in Japanese, 5 in Chinese, and 1 each in Norwegian, Polish, and German. There were only 3 trials of acupuncture for acute low back pain. These studies did not justify firm conclusions because of their small sample sizes and low methodologic quality. For chronic low back pain, there is evidence of pain relief and functional improvement for acupuncture compared to no treatment or sham therapy. These effects were only observed immediately after the end of the sessions and in short term follow-up. There is also evidence that acupuncture, added to other conventional therapies, relieves pain and improves function better than the conventional therapies alone. However, the effects are only small. Dry-needling appears to be a useful adjunct to other therapies for chronic low back pain. No clear recommendations could be made about the most effective acupuncture technique. CONCLUSIONS: The data do not allow firm conclusions regarding the effectiveness of acupuncture for acute low back pain. For chronic low back pain, acupuncture is more effective for pain relief and functional improvement than no treatment or sham treatment immediately after treatment and in the short-term only. Acupuncture is not more effective than other conventional and "alternative" treatments. The data suggest that acupuncture and dry-needling may be useful adjuncts to other therapies for chronic low back pain. Because most of the studies were of lower methodologic quality, there is a clear need for higher quality trials in this area. PMID- 15834339 TI - Long-term outcomes of surgical and nonsurgical management of lumbar spinal stenosis: 8 to 10 year results from the maine lumbar spine study. AB - STUDY DESIGN: A prospective observational cohort study. OBJECTIVE: To assess long term outcomes of patients with lumbar spinal stenosis treated surgically or nonsurgically. SUMMARY OF BACKGROUND DATA: The relative benefit of various treatments for lumbar spinal stenosis is uncertain. Surgical treatment has been associated with short-term improvement, but recurrence of symptoms has been documented. Few studies have compared long-term outcomes of surgical and nonsurgical treatments. METHODS: Patients recruited from the practices of orthopaedic surgeons, neurosurgeons, and occupational medicine physicians throughout Maine had baseline interviews with follow-up questionnaires mailed at regular intervals over 10 years. Clinical data were obtained at baseline from a physician questionnaire. Most patients initially undergoing surgery had a laminectomy without fusion performed. Outcomes including patient-reported symptoms of leg and back pain, functional status, and satisfaction were assessed at 8- to 10-year follow-up. Primary analyses were based on initial treatment received with secondary analyses examining actual treatment received by 10 years. RESULTS: Of 148 eligible consenting patients initially enrolled, 105 were alive after 10 years (67.7% survival rate). Among surviving patients, long-term follow up between 8 and 10 years was available for 97 of 123 (79%) patients (including 11 patients who died before the 10-year follow-up but completed a 8 or 9 year survey); 56 of 63 (89%) initially treated surgically and 41 of 60 (68%) initially treated nonsurgically. Patients undergoing surgery had worse baseline symptoms and functional status than those initially treated nonsurgically. Outcomes at 1 and 4 years favored initial surgical treatment. After 8 to 10 years, a similar percentage of surgical and nonsurgical patients reported that their low back pain was improved(53% vs. 50%, P = 0.8), their predominant symptom (either back or leg pain) was improved (54% vs. 42%, P = 0.3), and they were satisfied with their current status (55% vs. 49%, P = 0.5). These treatment group findings persisted after adjustment for other determinants of outcome in multivariate models. However, patients initially treated surgically reported less severe leg pain symptoms and greater improvement in back-specific functional status after 8 to 10 years than nonsurgically treated patients. By 10 years, 23% of surgical patients had undergone at least one additional lumbar spine operation, and 39% of nonsurgical patients had at least one lumbar spine operation. Patients undergoing subsequent surgical procedures had worse outcomes than those continuing with their initial treatment. Outcomes according to actual treatment received at 10 years did not differ because individuals undergoing additional surgical procedures had worse outcomes than those continuing with their initial treatment. CONCLUSIONS: Among patients with lumbar spinal stenosis completing 8- to 10-year follow-up, low back pain relief, predominant symptom improvement, and satisfaction with the current state were similar in patients initially treated surgically or nonsurgically. However, leg pain relief and greater back-related functional status continued to favor those initially receiving surgical treatment. These results support a shared decision-making approach among physicians and patients when considering treatment options for lumbar spinal stenosis. PMID- 15834341 TI - Measurement of fracture kyphosis with the Oxford Cobbometer: intra- and interobserver reliabilities and comparison with other techniques. AB - STUDY DESIGN: Statistical analysis of 3 techniques for measuring thoracolumbar kyphosis secondary to fracture. OBJECTIVES: To determine the reliability of using an Oxford Cobbometer and assess the most reliable measurement technique. SUMMARY OF BACKGROUND DATA: The reproducibility of Cobb angles for the assessment of saggital plane deformity on spine radiographs has been shown to have significant variability in both intra- and interobserver error. METHODS: Twenty-four lateral spine radiographs of patients with thoracic and lumbar vertebral fractures were measured on 2 separate occasions, in random order, by 4 blinded observers using the same Oxford Cobbometer and ruler. RESULTS: Method 2, the angle from the inferior endplate of the vertebra above the fractured vertebra to the superior endplate of the vertebra below the fractured vertebra, had the greatest intraobserver and interobserver reliabilities (rho = 0.856-0.976 and rho = 0.95, respectively). The other 2 methods had lower reliabilities; however, all 3 methods were well above the statistically acceptable threshold of >0.8, and the intraobserver reliabilities with each observer was 99% overall. These reliabilities supersede results reported previously using the conventional Cobb technique. The absolute mean difference between readings and 95% limit of agreement also improves on previous data, 2 degrees and +/- 5.8 degrees , respectively. CONCLUSIONS: Highest intraclass correlation coefficients were obtained using method 2. Using the Oxford Cobbometer to measure fracture kyphosis has higher reliability than the standard Cobb angle technique. It is easy and quick to use in a clinical setting. PMID- 15834342 TI - European comparison of costs and quality in the treatment of acute back pain. AB - STUDY DESIGN: Retrospective multicenter observational study. OBJECTIVES: To compare the outpatient quality and costs of treating acute back pain in England, Germany, the Netherlands, and Switzerland. SUMMARY OF BACKGROUND DATA: No study has yet attempted to compare the quality, costs, and resource utilization of acute back pain treatment in Europe. METHODS: A total of 130 randomly selected physician practices assessed services for 1 hypothetical average patient during the first 4 weeks of treatment (cost evaluation) and 127 practices reported retrospective data on 1 real patient (quality evaluation) in 2001. Reimbursement fees served as unit costs for Germany and Switzerland. Average reimbursement fees were used to measure resource utilization in all countries. Quality of care was assessed in terms of the following unnecessary treatments and diagnoses: bed rest for more than 2 days; exercise therapy; scheduling of a radiograph or other imaging tests; and referral to another provider. Responses were weighted with the level of scientific evidence for overuse. RESULTS: Weighted-average overuse ranged from 18% in the Netherlands to 31% in Germany. In England, Germany, and Switzerland, at least a third of the resources used to treat back pain were wasted. CONCLUSIONS: There was considerable waste in treating acute back pain. The Netherlands had highest quality and lowest resource utilization in providing treatment for acute back pain. PMID- 15834343 TI - Clinical course and prognostic factors in acute low back pain: patients consulting primary care for the first time. AB - STUDY DESIGN: Inception cohort study. OBJECTIVES: To examine the clinical course of acute low back pain and to evaluate prognostic factors for nonrecovery. SUMMARY OF BACKGROUND DATA: Few studies have explored clinical course and prognostic factors in patients who consult primary care for their first time because of an episode of low back pain of <3 weeks duration. METHODS: A total of 123 patients with acute low back pain <3 weeks consulting primary care for the first time were included, and 120 completed 3 months follow-up. Baseline assessments included sociodemographic characteristics, back pain history and current status, psychological questionnaires and clinical examination. Main outcome measures were pain intensity, disability by Roland Morris Disability Questionnaire, and recovery of disability. Potential prognostic factors for recovery or not were analyzed by multivariate logistic regression. RESULTS: At 4 weeks and 3 months 76% of the patients had recovered. Mean pain intensity and mean disability scores dropped 58% and 68%, respectively, of initial levels during the 3 months. The proportion with sickness absence was 8% at 4 weeks and 6% at 3 months. Several sociodemographic, clinical, and psychological factors were of prognostic value. Compared with their respective reference categories, age above 45 years (odds ratio 4.4, 95% confidence interval 1.4-14.0), smoking (3.0, 1.1-8.5), two or more neurological signs (4.6, 1.4-14.9), a score of >or=90 on the psychosocial screening (3.1, 1.0-9.4), and high levels of distress (4.1, 1.3-12.8) were the best prognostic factors of nonrecovery at 3 months. CONCLUSION: During a period of 3 months, 24% of the patients had not recovered. Psychological factors and neurological signs were strongly associated with nonrecovery at 3 months. In addition to the traditional examination of neurological symptoms and signs, psychological factors should be considered already at the initial visit of an episode of low back pain. PMID- 15834344 TI - Low back pain and pelvic pain during pregnancy: prevalence and risk factors. AB - STUDY DESIGN: Cross-sectional study. Women giving birth at one of two hospitals of northern Sweden from 1 January 2002 until 30 April 2002 were invited to fill in a questionnaire on their obstetric and gynecological history, actual pregnancy, and delivery. OBJECTIVE: The aim of this study was to investigate prevalence and risk factors for low back pain and pelvic pain (LBPP) during pregnancy. SUMMARY OF BACKGROUND DATA: Although low back pain and pelvic pain during pregnancy is a most common complication of pregnancy, its etiology is unknown and the pathophysiology is poorly understood. METHODS: The sample was analyzed by calculating the prevalence of LBPP during pregnancy. Univariate and multivariate logistic regression was performed to calculate odds ratio (OR) and its 95% confidence intervals (CI) where applicable. Parametric and nonparametric testing was used to establish differences between groups. RESULTS: The response rate was 83.2% (N = 891). The prevalence of LBPP during pregnancy was 72%. Most cases reported both anterior and posterior pain. Increasing parity, history of hypermobility, and reported periods of amenorrhea were risk factors for LBPP. Women with LBPP had significantly higher prepregnancy weight, end-pregnancy weight, and prepregnancy and end-pregnancy body mass index. Age at menarche and use of oral contraceptives were not associated with LBPP. Nonrespondents were of the same age and parity as respondents. CONCLUSIONS: A majority of pregnant women report LBPP. Parity, LBPP during a previous pregnancy, body mass index, a history of hypermobility, and amenorrhea are factors influencing the risk of developing LBPP during pregnancy. PMID- 15834345 TI - Spine MRI findings in a patient with tuberous sclerosis: a case report--part II. PMID- 15834346 TI - Transforming care. PMID- 15834347 TI - E-learning as educational tool in emergency and disaster medicine teaching. AB - E-learning is a new project for education based on the adoption of new computerised, multimedia and telematic technologies. Its application has deeply changed the concept of a teacher-based teaching to a student-centred educational project. It offers a great flexibility in the educational methodology, in the administration of contents, in the synchronous and/or asynchronous interaction between teachers and students, in the organisation and in the structure of the course, in the educational plans, in the support, the tracking and the evaluation of the student. E-learning could represent a great resource and a possible revolution in the concept of education and in the field of medical education as well. In some specific fields of application, as Emergency and Disaster Medicine, where the interaction between the student, the teacher and the patient, even if of great importance, are difficult to obtain in a quiet setting and have a lot of organizing, technical and economic troubles, e-learning approach could find a fertile field of application. In this paper we present a new program of educational activities we started in the field of Emergency and Disaster Medicine together with a review of the history of the instructional design and related technologies, ranging from the development of computer aided instruction to modern e-learning applications as teaching methodologies, and their impact on pedagogic and operative aspects. PMID- 15834348 TI - Effect of different sterile barrier precautions and central venous catheter dressing on the skin colonization around the insertion site. AB - AIM: The purpose of this prospective, randomized, controlled study was the comparison of maximal sterile barrier (consisting of mask, cap, sterile gloves, gown, large drape) vs control precautions (mask, cap, sterile gloves, small drape) and of transparent polyurethan film vs gauze dressing for use on central venous (CVC) nontunneled catheters, inserted via the jugular vein. Skin colonization at the insertion site (defined by quantitative skin cultures performed at the time of insertion and in days 2 and 5) was used as a primary endpoint. Catheter tip colonization was also assessed through qualitative culture and CVC related sepsis was defined by the isolation of the same organism from the catheter tip and the blood, with clinical sepsis of no other apparent source. METHODS: Eighty-two consecutive patients were enrolled, admitted to a mixed medical-surgical ICU, aged 72+/-12 years, 58% male, SAPS II 42+/-13. One-hundred and seven CVCs were studied (presenting 750 catheter in situ days); CVCs were in place for a mean period of 6.9+/-4.7 days and 5 episodes of central catheter related bloodstream infection were detected (6.6 per 1000 catheter days). RESULTS: A multiple logistic regression detected an increased risk of skin colonization in male gender (OR=2.5) and control precautions (OR=3.4) and no difference with regard to age, dressing and diagnostic group. CONCLUSIONS: Maximal sterile barrier proved to be an effective and recommended practice. However surveillance skin cultures revealed the common and changing nature of colonization of skin at the insertion site. PMID- 15834349 TI - Pancreas transplantation: problems and prospects in intensive care units. AB - AIM: The aim of this study is to describe personal experience in the intensive management of patients with severe diabetes undergoing pancreas transplantation. METHODS: Clinical records of subjects consecutively undergoing an isolated or combined pancreas transplant have been examinated. RESULTS: During the considered period, 10 patients received an isolated pancreas transplant and 43 a simultaneous kidney-pancreas transplantation (SPKT), including 6 using a kidney from a living donor. The mean stay in the Intensive Care Unit (ICU) was 4.7 days: 52 patients (98.2%) were transferred to the Surgical Department, whereas one (1.8%) belonging to the SPKT group died with a non-functioning graft. Ten patients (18.6%) were re-admitted because of the onset of late complications, including one SPKT who died of sudden cardiac death with functioning grafts. Arterial hypertension appeared in 51% of the recipients, and 5.6% experienced at least one hypotensive episode. Cardiac rhythm alterations were diagnosed in 5 subjects (9.4%), and myocardial ischemia in 9 (17%). CONCLUSIONS: Pancreas transplantation is a therapeutic option that can improve patients' quality of life by also slowing down the evolution of diabetes; however, it is important to bear in mind the associated risks. The best results are obtained in patients in whom the disease has not already seriously impaired the function of the various target organs. PMID- 15834350 TI - Femoral and sciatic nerves block (BiBlock) in orthopedic traumatologic lower limbs surgery in patients with multiple sclerosis. AB - Multiple sclerosis is a progressive demyelinating disease which affects large areas of the brain and of the spinal cord. Stressful events, surgical procedures, general anaesthesia and central blocks seem to be responsible for relapses, with worsening of the disease. So, when we scheduled 2 patients with multiple sclerosis for lower limbs orthopedic traumatologic surgery, we decided to use a peripheral block, and in particular a BiBlock. The patients' evaluation in the immediate postoperative course and 30 days after surgery has shown no relapses of the disease. In the literature, however, data about anaesthesia and multiple sclerosis are few and controversial, sometimes in contrast. Anyway, the use of peripheral blocks has neither anatomic, nor metabolic interferences with the lesion sites of multiple sclerosis. In conclusion, peripheral block is safe and it is the technique of choice for this type of patients, when surgery allows it. PMID- 15834351 TI - Different anesthesiological management in two high risk pregnant women with heart failure undergoing emergency cesarean section. AB - Pregnancy exacerbates heart diseases. The aim of this clinical report is to review the different anesthesiological management of emergent cesarean section in 2 patients with heart failure. The pathophysiology of heart failure is described according to the primary cause of disease, as well as the impact of 2 different anesthetic techniques. Two case reports of a university referral hospital are presented. Both patients left the hospital in good general conditions. Case 1: a pregnant patient with severe aortic regurgitation who received epidural anesthesia. Case 2: a pregnant patient with peripartum cardiomyopathy who was given general anesthesia. Medical and surgical therapies for aortic regurgitation and peripartum dilated cardiomyopathy are evolving. Adequate knowledge of anesthesiology is required to appropriately manage these cases. We tailored the anesthetic technique to the specific characteristics of our 2 patients. The beneficial effects of sympathectomy were observed in the postoperative period of case 1; the use of high doses opiates minimised dangerous cardiovascular changes in case 2, but rapid resuscitation of the baby should be available. Selection of the anesthetic technique in obstetrics is the most challenging issue for the anesthesiologist: extensive knowledge of the pathophysiology of heart disease is required for an optimal choice. PMID- 15834352 TI - Hypophosphatemia and metabolic acidosis. AB - The aim of the paper was to describe an unusual case of non lactic metabolic acidosis connected to hypophosphatemia and refractory to infusion of bicarbonate. A 37 year old man was admitted to Intensive Care Unit with a severe metabolic acidosis. On admission the arterial gas analysis showed non lactic metabolic acidosis (pH 7.17; base excess [BE] -20.3; lactic acid 0.8 mMol/L), with hypoxemia and critical hypocapnia. Despite therapy with bicarbonate the acidosis persisted. After 4 hours glucose phosphate was administered, although the phosphoremia was unknown. After phosphate supplementation an improvement of acidosis was observed. Our hypothesis is that in the kidney phosphate depletion caused impaired tubular reabsorption of bicarbonate, which led to a non lactic metabolic acidosis. PMID- 15834354 TI - Obesity-related cardiovascular risk factors: intervention recommendations to decrease adolescent obesity. AB - The incidence of adolescent obesity is increasing dramatically in the United States with associated risks of hypertension, adverse lipid profiles, and Type II diabetes. Unless reversed, this trend predicts an epidemic of adult cardiovascular disease. Interventions at home, at school, and in the community are required to empower teens to increase physical activity and to modify eating habits. This article describes assessment for obesity-related health problems as well as scientific guidelines and research-based intervention strategies to decrease obesity in adolescents. PMID- 15834355 TI - Quality health care for adolescents with special health-care needs: issues and clinical implications. AB - Assuring quality health care for the diverse U.S. adolescent population is a subject of growing concern among health-care providers and policy makers. Health care services and policies must appreciate that experiences of adolescents with special health-care needs include challenges that stretch beyond those unique to their conditions to include relationships with family, friends, and professionals as well as their ability to function in school, clinics, and other settings. This review explores those issues while building upon the six core outcomes delineated in the draft 10-year action plan offered as an accompaniment to the U.S. Department of Health and Human Services' Healthy People 2010 guidelines. PMID- 15834356 TI - An analysis of health promotion and risk behaviors of freshman college students in a rural southern setting. AB - This study described health promotion and risk behaviors of entering college students in a rural southern setting. A sample of 251 students completed the Health Promotion and Health Risk Survey during a testing period that included other academic surveys. Results indicated that participants believe they are in control of their health. In addition, participants reported that happiness and health were the most important personal values for them. When compared with the National College Health Risk Behavior data, participants in this study were significantly lower in many of the health risk behaviors. PMID- 15834357 TI - Creating an international nursing practice and education workplace. AB - In today's global workplace, both nursing practice and education need to promote international health. Nurses from a comprehensive 373-patient-bed hospital at Cincinnati Children's Hospital Medical Center and faculty from the College of Nursing, University of Cincinnati, in southwest Ohio have successfully collaborated to develop several unique international nursing exchange programs. The goals of these programs are to increase cultural sensitivity and nursing knowledge relevant to a global community. The essential components used in creating and implementing the programs with Scotland, Honduras, and Korea will serve as an international workplace model for others, especially for those settings focused on children and family health care. PMID- 15834358 TI - Babies breathing. PMID- 15834359 TI - Attention deficit/hyperactivity disorder: working the system for better diagnosis and treatment. PMID- 15834361 TI - Breastfeeding initiation and duration in Lebanon: are the hospitals "mother friendly"? AB - Eight hundred and thirty mothers and their children aged 1 to 5 years were studied to identify the determinants of breastfeeding initiation and perseverance in relation to hospital policies. Only 18.3% of the mothers initiated breastfeeding within half an hour after birth. Mothers whose baby was brought to them every 3 hours or less were 2.5 times and mothers whose infants were brought to them for night feedings were 3 times more likely to initiate breastfeeding a few hours after birth. Mother's occupational status and whether she was breastfed were significantly associated with breastfeeding duration. PMID- 15834362 TI - A brief report: from meeting room to telecommunication studio. PMID- 15834378 TI - A descriptive study of heavy emergency department users at an academic emergency department reveals heavy ED users have better access to care than average users. AB - INTRODUCTION: Emergency department (ED) overcrowding has been a significant problem for the last 10 years. Several studies have shown that a relatively small number of ED patients are responsible for a disproportionate amount of ED visits. This study aims to describe the frequent users of our emergency department. METHODS: This was an institutional review board-approved descriptive study performed by a retrospective review of electronic records. This pilot describes and compares patients who had 12 or more ED visits during the study year with those who visited less. RESULTS: The 234 patients who met criteria for high frequency use (HFU) of the emergency department were responsible for a total of 4633 visits. Sex, race, and age distribution of HFU patients were similar to those of general ED patients. Eighty-four percent of HF users have insurance and 93% have primary care providers. A relatively small percentage of HFU visits, 4%, were mental health-related visits and 3% were alcohol- and drug-related visits. The HFU visits are socially connected: 93% have their own homes; 94% have relatives or friends; 73% have a religious affiliation. Pain or pain-related conditions are the most common diagnoses. These patients are also frequent users of ambulatory care services. CONCLUSION: The similarities between our HFU and the general ED population are more numerous than their differences. The HFU patients of our emergency department are different in terms of age, employment status, and type of insurance. IMPLICATIONS FOR NURSES: A detailed description of local HFU may help to inform planning and better meet ED patients' needs. As one of many results of this study, the ED chairman met with the Hematology-Oncology team and reviewed the protocol for ED management of sickle cell crisis. The meeting resulted in a revised protocol, including an immediate change in their pain medication from meperidine to either morphine or hydromorphone. PMID- 15834379 TI - Primary reasons for ED visits and procedures performed for patients who saw nurse practitioners. AB - INTRODUCTION: Little is known, from a national perspective, about what types of patients are seen by nurse practitioners in the emergency department. METHODS: Data from 1545 participating emergency departments across the United States during 1997, 1998, 1999, and 2000 were collected from nationally representative samples of urban and rural hospitals using the National Hospital Ambulatory Medical Care Surveys. Results Nurse practitioners saw 5.76 million ED patients during the 4-year period. Using the Reason for Visit Classification developed by the National Center for Health Statistics, the primary category for patients seen by nurse practitioners was classified as "Injury by type and/or location." The types of injuries in this category were lacerations and cuts to an upper extremity and facial area; injuries to the head, neck, and face; and foreign bodies in the eye. The next most common category was classified under "General symptoms." Nurse practitioners saw patients in this category with symptoms of chest pain, side or flank pain, fever, and edema. DISCUSSION: The findings from this study provide insight into the types of patient visits seen by nurse practitioners in emergency departments in the United States and the services and procedures that were received by patients. PMID- 15834380 TI - What is needed to assist patients with advance directives from the perspective of emergency nurses. AB - INTRODUCTION: Nurses, by the nature of their practice, are in a unique position to help patients complete advance directives (ADs). This article focuses on an open-ended question that was part of a larger quantitative survey, "What do emergency nurses need to increase their ability to assist patients with ADs?" METHODS: A random sample of emergency nurses in four states was surveyed. Of the 663 nurses who responded to the survey, 506 (76%) wrote responses to the open ended question. Thematic analysis was used to establish and saturate themes. RESULTS: The major themes discussed most often by the nurses were the need for more education and more time to discuss ADs with patients. Other themes discussed by nurses were the need for: a supportive work environment, other health care professionals' participation conducive to assisting patients, patient/public education related to ADs, communication, and particular nursing characteristics needed to successfully assist patients with ADs. Not all the nurses in the study felt that assisting patients with ADs was the role of emergency nurses. CONCLUSIONS: Those nurses who saw assisting patients with ADs as their role were very clear about what they needed to be able to carry out the role (more time to spend with patients and more education related to ADs) and that they could not fulfill that role without the support of their institution's administration. PMID- 15834381 TI - Irrigating simple acute traumatic wounds: a review of the current literature. PMID- 15834382 TI - The use of a mental health triage assessment tool in a busy Canadian tertiary care children's hospital. PMID- 15834383 TI - Hospital-based intervention for intimate partner violence victims: a forensic nursing model. PMID- 15834385 TI - Designing a rabies postexposure prophylaxis program with emphasis on staff and patient education. PMID- 15834386 TI - Inadvertent attachment of a blood pressure device to a needleless IV "Y-site": surprising, fatal connections. PMID- 15834387 TI - The Notebook, revisited. PMID- 15834388 TI - Emergency nurse urges booster seat advocacy after encounter at traumatic crash scene. PMID- 15834391 TI - A 23-year-old man with chest pressure, pallor, tachypnea, and tonsillitis. PMID- 15834392 TI - Emergency Nursing Pediatric Course (ENPC): the new 3rd Edition. PMID- 15834393 TI - Confusion, somnolence, seizures, tachycardia? Question drug-induced hypoglycemia. PMID- 15834394 TI - Policy advocacy for children. PMID- 15834395 TI - Penetrating cardiac trauma: a case study. PMID- 15834396 TI - A 61-year-old man with a self-diagnosed back injury and difficulty walking: be suspicious, ask the question. PMID- 15834397 TI - Medical mission to Bolivia: a photo essay. PMID- 15834403 TI - Stable antibody expression at therapeutic levels using the 2A peptide. AB - Therapeutic monoclonal antibodies (mAbs) are currently being developed for the treatment of cancer and other diseases. Despite clinical success, widespread application of mAb therapies may be limited by manufacturing capabilities. In this paper, we describe a mAb delivery system that allows continuous production of a full-length antibody at high-concentrations in vivo after gene transfer. The mAb is expressed from a single open reading frame by linking the heavy and light chains with a 2A self-processing peptide derived from the foot-and-mouth disease virus. Using this expression system, we generated a recombinant adeno-associated virus vector encoding the VEGFR2-neutralizing mAb DC101 (rAAV8-DC101). A single dose of rAAV8-DC101 resulted in long-term expression of >1,000 microg/ml of DC101 in mice, demonstrating significant anti-tumor efficacy. This report describes the first feasible gene therapy approach for stable delivery of mAbs at therapeutic levels, which may serve as an attractive alternative to direct injection of mAbs. PMID- 15834404 TI - Quantitative mouse brain proteomics using culture-derived isotope tags as internal standards. AB - An important challenge for proteomics is to be able to compare absolute protein levels across biological samples. Here we introduce an approach based on the use of culture-derived isotope tags (CDITs) for quantitative tissue proteome analysis. We cultured Neuro2A cells in a stable isotope-enriched medium and mixed them with mouse brain samples to serve as internal standards. Using CDITs, we identified and quantified a total of 1,000 proteins, 97-98% of which were expressed in both mouse whole brain and Neuro2A cells. CDITs also allow comprehensive and absolute protein quantification. Synthetic unlabeled peptides were used to quantify the corresponding proteins labeled with stable isotopes in Neuro2A cells, and the results were used to obtain the absolute amounts of 103 proteins in mouse whole brain. The expression levels correlated well with those in Neuro2A cells. Thus, the use of CDITs allows both relative and absolute quantitative proteome studies. PMID- 15834405 TI - In vivo detection of amyloid-beta deposits by near-infrared imaging using an oxazine-derivative probe. AB - As Alzheimer's disease pathogenesis is associated with the formation of insoluble aggregates of amyloid beta-peptide, approaches allowing the direct, noninvasive visualization of plaque growth in vivo would be beneficial for biomedical research. Here we describe the synthesis and characterization of the near infrared fluorescence oxazine dye AOI987, which readily penetrates the intact blood-brain barrier and binds to amyloid plaques. Using near-infrared fluorescence imaging, we demonstrated specific interaction of AOI987 with amyloid plaques in APP23 transgenic mice in vivo, as confirmed by postmortem analysis of brain slices. Quantitative analysis revealed increasing fluorescence signal intensity with increasing plaque load of the animals, and significant binding of AOI987 was observed for APP23 transgenic mice aged 9 months and older. Thus, AOI987 is an attractive probe to noninvasively monitor disease progression in animal models of Alzheimer disease and to evaluate effects of potential Alzheimer disease drugs on the plaque load. PMID- 15834406 TI - Uncontrolled calcium sparks act as a dystrophic signal for mammalian skeletal muscle. AB - Most excitable cells maintain tight control of intracellular Ca(2+) through coordinated interaction between plasma membrane and endoplasmic or sarcoplasmic reticulum. Quiescent sarcoplasmic reticulum Ca(2+) release machinery is essential for the survival and normal function of skeletal muscle. Here we show that subtle membrane deformations induce Ca(2+) sparks in intact mammalian skeletal muscle. Spontaneous Ca(2+) sparks can be reversibly induced by osmotic shock, and participate in a normal physiological response to exercise. In dystrophic muscle with fragile membrane integrity, stress-induced Ca(2+) sparks are essentially irreversible. Moreover, moderate exercise in mdx muscle alters the Ca(2+) spark response. Thus, membrane-deformation-induced Ca(2+) sparks have an important role in physiological and pathophysiological regulation of Ca(2+) signalling, and uncontrolled Ca(2+) spark activity in connection with chronic activation of store operated Ca(2+) entry may function as a dystrophic signal in mammalian skeletal muscle. PMID- 15834407 TI - Suppression of HIV-1 infection by a small molecule inhibitor of the ATM kinase. AB - Chemotherapy that is used to treat human immunodeficiency virus type-1 (HIV-1) infection focuses primarily on targeting virally encoded proteins. However, the combination of a short retroviral life cycle and high mutation rate leads to the selection of drug-resistant HIV-1 variants. One way to address this problem is to inhibit non-essential host cell proteins that are required for viral replication. Here we show that the activity of HIV-1 integrase stimulates an ataxia telangiectasia-mutated (ATM)-dependent DNA damage response, and that a deficiency of this ATM kinase sensitizes cells to retrovirus-induced cell death. Consistent with these observations, we demonstrate that a novel and specific small molecule inhibitor of ATM kinase activity, KU-55933, is capable of suppressing the replication of both wild-type and drug-resistant HIV-1. PMID- 15834408 TI - The KIF3 motor transports N-cadherin and organizes the developing neuroepithelium. AB - In the developing brain, the organization of the neuroepithelium is maintained by a critical balance between proliferation and cell-cell adhesion of neural progenitor cells. The molecular mechanisms that underlie this are still largely unknown. Here, through analysis of a conditional knockout mouse for the Kap3 gene, we show that post-Golgi transport of N-cadherin by the KIF3 molecular motor complex is crucial for maintaining this balance. N-cadherin and beta-catenin associate with the KIF3 complex by co-immunoprecipitation, and colocalize with KIF3 in cells. Furthermore, in KAP3-deficient cells, the subcellular localization of N-cadherin was disrupted. Taken together, these results suggest a potential tumour-suppressing activity for this molecular motor. PMID- 15834409 TI - Lymphocyte arrest requires instantaneous induction of an extended LFA-1 conformation mediated by endothelium-bound chemokines. AB - It is widely believed that rolling lymphocytes require successive chemokine induced signaling for lymphocyte function-associated antigen 1 (LFA-1) to achieve a threshold avidity that will mediate lymphocyte arrest. Using an in vivo model of lymphocyte arrest, we show here that LFA-1-mediated arrest of lymphocytes rolling on high endothelial venules bearing LFA-1 ligands and chemokines was abrupt. In vitro flow chamber models showed that endothelium-presented but not soluble chemokines triggered instantaneous extension of bent LFA-1 in the absence of LFA-1 ligand engagement. To support lymphocyte adhesion, this extended LFA-1 conformation required immediate activation by its ligand, intercellular adhesion molecule 1. These data show that chemokine-triggered lymphocyte adhesiveness involves a previously unrecognized extension step that primes LFA-1 for ligand binding and firm adhesion. PMID- 15834410 TI - Egr-2 and Egr-3 are negative regulators of T cell activation. AB - T cell receptor engagement in the absence of proper accessory signals leads to T cell anergy. E3 ligases are involved in maintaining the anergic state. However, the specific molecules responsible for the induction of anergy have yet to be elucidated. Using microarray analysis we have identified here early growth response gene 2 (Egr-2) and Egr-3 as key negative regulators of T cell activation. Overexpression of Egr2 and Egr3 was associated with an increase in the E3 ubiquitin ligase Cbl-b and inhibition of T cell activation. Conversely, T cells from Egr3(-/-) mice had lower expression of Cbl-b and were resistant to in vivo peptide-induced tolerance. These data support the idea that Egr-2 and Egr-3 are involved in promoting a T cell receptor-induced negative regulatory genetic program. PMID- 15834411 TI - Direct visualization of dislocation dynamics in grain-boundary scars. AB - Mesoscale objects with unusual structural features may serve as the analogues of atoms in the design of larger-scale materials with novel optical, electronic or mechanical behaviour. In this paper we investigate the structural features and the equilibrium dynamics of micrometre-scale spherical crystals formed by polystyrene particles adsorbed on the surface of a spherical water droplet. The ground state of sufficiently large crystals possesses finite-length grain boundaries (scars). We determine the elastic response of the crystal by measuring single-particle diffusion, and quantify the fluctuations of individual dislocations about their equilibrium positions within a scar by determining the dislocation spring constants. We observe rapid dislocation glide with fluctuations over the barriers separating one local Peierls minimum from the next and rather weak binding of dislocations to their associated scars. The long distance (renormalized) dislocation diffusion glide constant is extracted directly from the experimental data and is found to be moderately faster than single-particle diffusion. We are also able to determine the parameters of the Peierls potential induced by the underlying crystalline lattice. PMID- 15834412 TI - Evolutionary approach for determining first-principles hamiltonians. AB - Modern condensed-matter theory from first principles is highly successful when applied to materials of given structure-type or restricted unit-cell size. But this approach is limited where large cells or searches over millions of structure types become necessary. To treat these with first-principles accuracy, one 'coarse-grains' the many-particle Schrodinger equation into 'model hamiltonians' whose variables are configurational order parameters (atomic positions, spin and so on), connected by a few 'interaction parameters' obtained from a microscopic theory. But to construct a truly quantitative model hamiltonian, one must know just which types of interaction parameters to use, from possibly 10(6)-10(8) alternative selections. Here we show how genetic algorithms, mimicking biological evolution ('survival of the fittest'), can be used to distil reliable model hamiltonian parameters from a database of first-principles calculations. We demonstrate this for a classic dilemma in solid-state physics, structural inorganic chemistry and metallurgy: how to predict the stable crystal structure of a compound given only its composition. The selection of leading parameters based on a genetic algorithm is general and easily applied to construct any other type of complex model hamiltonian from direct quantum-mechanical results. PMID- 15834413 TI - Supramolecular barrels from amphiphilic rigid-flexible macrocycles. AB - Precise control of supramolecular objects requires the rational design of molecular components, because the information determining their specific assembly should be encoded in their molecular architecture. In this context, diverse self assembling molecules including liquid crystals, dendrimers, block copolymers, hydrogen-bonded complexes and rigid macrocycles are being created as a means of manipulating supramolecular structure. Incorporation of a stiff rod-like building block into an amphiphilic molecular architecture leads to another class of self assembling molecules. Aggregation of rod building blocks can generate various nanoscale objects including bundles, ribbons, tubules and vesicles, depending on the molecular structure and/or the presence of a selective solvent. We present here an unusual example of supramolecular barrels in the solid and in aqueous solution, based on the self-assembly of amphiphilic rigid-flexible macrocycles driven by non-covalent interactions. Preliminary experiments show that these amphiphilic macrocycles are membrane-active. The amphiphilic macrocycles might thus lead to an excellent model system for exploring biological processes in supramolecular materials. PMID- 15834414 TI - Vibrational coherence of self-organized silver nanocrystals in f.c.c. supra crystals. AB - Fabrication of devices from inorganic nanocrystals normally requires that they are self-organized into ordered structures. It has now been demonstrated that nanocrystals are able to self-organize in a 'supra'-crystal with a face-centred cubic (f.c.c.) structure. The physical properties of nanocrystals self-organized into compact arrays are quite different from those of both isolated nanocrystals and the bulk phase. The collective optical and magnetic properties of these nanocrystal assemblies are governed mainly by dipolar interactions. Here, we show that nanocrystals vibrate coherently when they are self-organized in f.c.c. supra crystals. Hence, a phase relation exists between the vibrations of all of the nanocrystals in a supra-crystal. This vibrational coherence can be observed by a substantial change of the quadrupolar low-frequency Raman scattering peak. Although a change in electronic transport properties has previously been observed on self-organization of silver nanocrystals, vibrational coherence represents the first intrinsic property of f.c.c. supra-crystals. PMID- 15834415 TI - Fabrication of reconfigurable protein matrices by cracking. AB - The interface between extracellular matrices and cells is a dynamic environment that is crucial for regulating important cellular processes such as signal transduction, growth, differentiation, motility and apoptosis. In vitro cellular studies and the development of new biomaterials would benefit from matrices that allow reversible modulation of the cell adhesive signals at a scale that is commensurate with individual adhesion complexes. Here, we describe the fabrication of substrates containing arrays of cracks in which cell-adhesive proteins are selectively adsorbed. The widths of the cracks (120-3,200 nm) are similar in size to individual adhesion complexes (typically 500-3,000 nm) and can be modulated by adjusting the mechanical strain applied to the substrate. Morphology of cells can be reversibly manipulated multiple times through in situ adjustment of crack widths and hence the amount of the cell-adhesive proteins accessible to the cell. These substrates provide a new tool for assessing cellular responses associated with exposure to matrix proteins. PMID- 15834416 TI - Nanoscale ferroelectric field-effect writing and reading using scanning tunnelling spectroscopy. AB - Control of the density of mobile charge carriers using electric fields is widely used in a variety of metal-insulator-semiconductor structures and is the governing principle behind the operation of field-effect transistors. Ferroelectric materials possessing a switchable and non-volatile polarization field can be used as insulating layers, revealing new opportunities for device applications. Advances in material processing and in particular complex oxide thin-film growth mean that high-quality field-effect devices can be based on ferroelectric/metallic oxide heterostructures. In addition, advances in local probe techniques such as atomic force microscopy allow them to be used in the imaging and study of small ferroelectric domain structures in bulk crystals and thin films. Meanwhile, scanning tunnelling microscopy and spectroscopy have established themselves as powerful techniques for atomic manipulation and nanometre-resolution electron tunnelling spectroscopy. Here, a scanning tunnelling microscope is used to investigate the ferroelectric field effect in all-perovskite heterostructures. Scanning tunnelling spectroscopy allows us to probe the local electronic properties of the polarized channel of a ferroelectric field-effect device as a function of the field orientation. This technique can be used to read and write ferroelectric field-induced regions with a size as low as 20 nm. PMID- 15834417 TI - Neurons in macaque area V4 acquire directional tuning after adaptation to motion stimuli. AB - Neurons in area V4 of the macaque are generally not selective for direction of motion, as judged from their response to directional stimuli presented after a baseline condition devoid of movement. We used motion adaptation to investigate whether stimulation history influences direction-of-motion selectivity. We found that classically nondirectional V4 neurons develop direction-of-motion selectivity after adaptation, an observation that underscores the dynamic nature of functional cortical architecture. PMID- 15834418 TI - Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. AB - Alpha-synuclein is phosphorylated at serine 129 (Ser129) in intracellular protein aggregates called Lewy bodies. These inclusion bodies are the characteristic pathologic lesions of Parkinson disease. Here we define the role of phosphorylation of Ser129 in alpha-synuclein toxicity and inclusion formation using a Drosophila model of Parkinson disease. Mutation of Ser129 to alanine to prevent phosphorylation completely suppresses dopaminergic neuronal loss produced by expression of human alpha-synuclein. In contrast, altering Ser129 to the negatively charged residue aspartate, to mimic phosphorylation, significantly enhances alpha-synuclein toxicity. The G protein-coupled receptor kinase 2 (Gprk2) phosphorylates Ser129 in vivo and enhances alpha-synuclein toxicity. Blocking phosphorylation at Ser129 substantially increases aggregate formation. Thus Ser129 phosphorylation status is crucial in mediating alpha-synuclein neurotoxicity and inclusion formation. Because increased number of inclusion bodies correlates with reduced toxicity, inclusion bodies may protect neurons from alpha-synuclein toxicity. PMID- 15834419 TI - Asymmetric membrane ganglioside sialidase activity specifies axonal fate. AB - Axon specification triggers the polarization of neurons and requires the localized destabilization of filamentous actin. Here we show that plasma membrane ganglioside sialidase (PMGS) asymmetrically accumulates at the tip of one neurite of the unpolarized rat neuron, inducing actin instability. Suppressing PMGS activity blocks axonal generation, whereas stimulating it accelerates the formation of a single (not several) axon. PMGS induces axon specification by enhancing TrkA activity locally, which triggers phosphatidylinositol-3-kinase (PI3K)- and Rac1-dependent inhibition of RhoA signaling and the consequent actin depolymerization in one neurite only. Thus, spatial restriction of an actin regulating molecular machinery, in this case a membrane enzymatic activity, before polarization is enough to determine axonal fate. PMID- 15834420 TI - The MAPK pathway and Egr-1 mediate stress-related behavioral effects of glucocorticoids. AB - Many of the behavioral consequences of stress are mediated by the activation of the glucocorticoid receptor by stress-induced high levels of glucocorticoid hormones. To explore the molecular mechanisms of these effects, we combined in vivo and in vitro approaches. We analyzed mice carrying a brain-specific mutation (GR(NesCre)) in the glucocorticoid receptor gene (GR, also called Nr3c1) and cell lines that either express endogenous glucocorticoid receptor or carry a constitutively active form of the receptor (DeltaGR) that can be transiently induced. In the hippocampus of the wild-type [corrected] mice after stress, as well as in the cell lines, activation of glucocorticoid receptors greatly increased the expression and enzymatic activity of proteins in the MAPK signaling pathway and led to an increase in the levels of both Egr-1 mRNA and protein. In parallel, inhibition of the MAPK pathway within the hippocampus abolished the increase in contextual fear conditioning induced by glucocorticoids. The present results provide a molecular mechanism for the stress-related effects of glucocorticoids on fear memories. PMID- 15834422 TI - Functional visualization of viral molecular motor by hydrogen-deuterium exchange reveals transient states. AB - Molecular motors undergo cyclical conformational changes and convert chemical energy into mechanical work. The conformational dynamics of a viral packaging motor, the hexameric helicase P4 of dsRNA bacteriophage phi8, was visualized by hydrogen-deuterium exchange and high-resolution mass spectrometry. Concerted changes of exchange kinetics revealed a cooperative unit that dynamically links ATP-binding sites and the central RNA-binding channel. The cooperative unit is compatible with a structure-based model in which translocation is mediated by a swiveling helix. Deuterium labeling also revealed the transition state associated with RNA loading, which proceeds via opening of the hexameric ring. The loading mechanism is similar to that of other hexameric helicases. Hydrogen-deuterium exchange provides an important link between time-resolved spectroscopic observations and high-resolution structural snapshots of molecular machines. PMID- 15834421 TI - Gbetagamma acts at the C terminus of SNAP-25 to mediate presynaptic inhibition. AB - Presynaptic inhibition mediated by G protein-coupled receptors may involve a direct interaction between G proteins and the vesicle fusion machinery. The molecular target of this pathway is unknown. We demonstrate that Gbetagamma mediated presynaptic inhibition in lamprey central synapses occurs downstream from voltage-gated Ca(2+) channels. Using presynaptic microinjections of botulinum toxins (BoNTs) during paired recordings, we find that cleavage of synaptobrevin in unprimed vesicles leads to an eventual exhaustion of synaptic transmission but does not prevent Gbetagamma-mediated inhibition. In contrast, cleavage of the C-terminal nine amino acids of the 25 kDa synaptosome-associated protein (SNAP-25) by BoNT A prevents Gbetagamma-mediated inhibition. Moreover, a peptide containing the region of SNAP-25 cleaved by BoNT A blocks the Gbetagamma inhibitory effect. Finally, removal of the last nine amino acids of the C terminus of SNAP-25 weakens Gbetagamma interactions with soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) complexes. Thus, the C terminus of SNAP-25, which links synaptotagmin I to the SNARE complex, may represent a target of Gbetagamma for presynaptic inhibition. PMID- 15834423 TI - CtBP represses p300-mediated transcriptional activation by direct association with its bromodomain. AB - Histone acetyltransferase coactivators bind to acetylated histones through their bromodomains and catalyze the acetylation of histone H3 and H4 tails for transcriptional activation. C-terminal binding protein (CtBP) serves as a transcriptional corepressor by recruiting histone deacetylases. However, the precise mechanism by which CtBP represses transcription has not been determined. In this study we found that CtBP1 directly associates with p300 by binding to the PXDLS motif in the bromodomain of p300. Moreover, CtBP1 blocks the accessibility of p300 to histones in an NADH-sensitive manner and thus represses p300-mediated histone acetylation and transcriptional activation. In addition, an NADH nonresponsive, monomeric mutant, CtBP1 (G183V), was found to strongly repress p300-mediated transcriptional activation. Thus, the dissociation of NADH from CtBP1 leads to the repression of p300-driven general transcriptional activity by CtBP1. These results suggest a novel mechanism whereby CtBP1 serves as an energy sensing repressor of histone acetyltransferase(s) and thus affects general transcription. PMID- 15834424 TI - The Hop2 and Mnd1 proteins act in concert with Rad51 and Dmc1 in meiotic recombination. AB - During the first meiotic division, homologous chromosomes (homologs) have to separate to opposite poles of the cell to ensure the right complement in the progeny. Homologous recombination provides a mechanism for a genome-wide homology search and physical linkage among the homologs before their orderly segregation. Rad51 and Dmc1 recombinases are the major players in these processes. Disruption of meiosis-specific HOP2 or MND1 genes leads to severe defects in homologous synapsis and an early-stage recombination failure resulting in sterility. Here we show that mouse Hop2 can efficiently form D-loops, the first recombination intermediates, but this activity is abrogated upon association with Mnd1. Furthermore, the Hop2-Mnd1 heterodimer physically interacts with Rad51 and Dmc1 recombinases and stimulates their activity up to 35-fold. Our data reveal an interplay among Hop2, Mnd1 and Rad51 and Dmc1 in the formation of the first recombination intermediates during meiosis. PMID- 15834425 TI - Integrin alphavbeta3 is a coreceptor for human cytomegalovirus. AB - Human cytomegalovirus (HCMV) is a widespread opportunistic pathogen that causes birth defects in newborns and severe disease in immunocompromised individuals. The broad tropism of HCMV infection suggests that it uses multiple receptors. We recently showed that the epidermal growth factor receptor (EGFR) serves as a receptor for HCMV. Here we show that HCMV also uses integrin alphavbeta3 as a coreceptor. Upon infection, HCMV glycoproteins gB and gH independently bind to EGFR and alphavbeta3, respectively, to initiate viral entry and signaling. Alphavbeta3 then translocates to lipid rafts where it interacts with EGFR to induce coordinated signaling. The coordination between EGFR and alphavbeta3 is essential for the early events of HCMV infection, including viral entry, RhoA downregulation, stress-fiber disassembly and viral nuclear trafficking. Our findings support a model in which EGFR and alphavbeta3 work together as coreceptors for HCMV entry and signaling. This discovery is fundamental to understanding HCMV pathogenesis and developing treatment strategies targeted to viral receptors. PMID- 15834427 TI - Amyloid beta protein immunotherapy neutralizes Abeta oligomers that disrupt synaptic plasticity in vivo. AB - One of the most clinically advanced forms of experimental disease-modifying treatment for Alzheimer disease is immunization against the amyloid beta protein (Abeta), but how this may prevent cognitive impairment is unclear. We hypothesized that antibodies to Abeta could exert a beneficial action by directly neutralizing potentially synaptotoxic soluble Abeta species in the brain. Intracerebroventricular injection of naturally secreted human Abeta inhibited long-term potentiation (LTP), a correlate of learning and memory, in rat hippocampus in vivo but a monoclonal antibody to Abeta completely prevented the inhibition of LTP when injected after Abeta. Size fractionation showed that Abeta oligomers, not monomers or fibrils, were responsible for inhibiting LTP, and an Abeta antibody again prevented such inhibition. Active immunization against Abeta was partially effective, and the effects correlated positively with levels of antibodies to Abeta oligomers. The ability of exogenous and endogenous antibodies to rapidly neutralize soluble Abeta oligomers that disrupt synaptic plasticity in vivo suggests that treatment with such antibodies might show reversible cognitive deficits in early Alzheimer disease. PMID- 15834426 TI - Diverse compounds mimic Alzheimer disease-causing mutations by augmenting Abeta42 production. AB - Increased Abeta42 production has been linked to the development of Alzheimer disease. We now identify a number of compounds that raise Abeta42. Among the more potent Abeta42-raising agents identified are fenofibrate, an antilipidemic agent, and celecoxib, a COX-2-selective NSAID. Many COX-2-selective NSAIDs tested raised Abeta42, including multiple COX-2-selective derivatives of two Abeta42-lowering NSAIDs. Compounds devoid of COX activity and the endogenous isoprenoids FPP and GGPP also raised Abeta42. These compounds seem to target the gamma-secretase complex, increasing gamma-secretase-catalyzed production of Abeta42 in vitro. Short-term in vivo studies show that two Abeta42-raising compounds increase Abeta42 levels in the brains of mice. The elevations in Abeta42 by these compounds are comparable to the increases in Abeta42 induced by Alzheimer disease causing mutations in the genes encoding amyloid beta protein precursor and presenilins, raising the possibility that exogenous compounds or naturally occurring isoprenoids might increase Abeta42 production in humans. PMID- 15834428 TI - Lactadherin promotes VEGF-dependent neovascularization. AB - Vascular endothelial growth factor (VEGF)-induced blood vessel growth is involved in both physiological and pathological angiogenesis and requires integrin mediated signaling. We now show that an integrin-binding protein initially described in milk-fat globule, MFG-E8 (also known as lactadherin), is expressed in and around blood vessels and has a crucial role in VEGF-dependent neovascularization in the adult mouse. Using neutralizing antibodies and lactadherin-deficient animals, we show that lactadherin interacts with alphavbeta3 and alphavbeta5 integrins and alters both VEGF-dependent Akt phosphorylation and neovascularization. In the absence of VEGF, lactadherin administration induced alphavbeta3- and alphavbeta5-dependent Akt phosphorylation in endothelial cells in vitro and strongly improved postischemic neovascularization in vivo. These results show a crucial role for lactadherin in VEGF-dependent neovascularization and identify lactadherin as an important target for the modulation of neovascularization. PMID- 15834429 TI - PI 3-kinase p110beta: a new target for antithrombotic therapy. AB - Platelet activation at sites of vascular injury is essential for the arrest of bleeding; however, excessive platelet accumulation at regions of atherosclerotic plaque rupture can result in the development of arterial thrombi, precipitating diseases such as acute myocardial infarction and ischemic stroke. Rheological disturbances (high shear stress) have an important role in promoting arterial thrombosis by enhancing the adhesive and signaling function of platelet integrin alpha(IIb)beta(3) (GPIIb-IIIa). In this study we have defined a key role for the Type Ia phosphoinositide 3-kinase (PI3K) p110beta isoform in regulating the formation and stability of integrin alpha(IIb)beta(3) adhesion bonds, necessary for shear activation of platelets. Isoform-selective PI3K p110beta inhibitors have been developed which prevent formation of stable integrin alpha(IIb)beta(3) adhesion contacts, leading to defective platelet thrombus formation. In vivo, these inhibitors eliminate occlusive thrombus formation but do not prolong bleeding time. These studies define PI3K p110beta as an important new target for antithrombotic therapy. PMID- 15834430 TI - Thromboxane A2 and prostaglandin F2alpha mediate inflammatory tachycardia. AB - Systemic inflammation induces various adaptive responses including tachycardia. Although inflammation-associated tachycardia has been thought to result from increased sympathetic discharge caused by inflammatory signals of the immune system, definitive proof has been lacking. Prostanoids, including prostaglandin (PG) D(2), PGE(2), PGF(2alpha), PGI(2) and thromboxane (TX) A(2), exert their actions through specific receptors: DP, EP (EP(1), EP(2), EP(3), EP(4)), FP, IP and TP, respectively. Here we have examined the roles of prostanoids in inflammatory tachycardia using mice that lack each of these receptors individually. The TXA(2) analog I-BOP and PGF(2alpha) each increased the beating rate of the isolated atrium of wild-type mice in vitro through interaction with TP and FP receptors, respectively. The cytokine-induced increase in beating rate was markedly inhibited in atria from mice lacking either TP or FP receptors. The tachycardia induced in wild-type mice by injection of lipopolysaccharide (LPS) was greatly attenuated in TP-deficient or FP-deficient mice and was completely absent in mice lacking both TP and FP. The beta-blocker propranolol did not block the LPS-induced increase in heart rate in wild-type animals. Our results show that inflammatory tachycardia is caused by a direct action on the heart of TXA(2) and PGF(2alpha) formed under systemic inflammatory conditions. PMID- 15834431 TI - Two distinct mechanisms for loss of thioredoxin-binding protein-2 in oxidative stress-induced renal carcinogenesis. AB - Thioredoxin is a major component of thiol-reducing system. Recently, we identified thioredoxin-binding protein-2 (TBP-2) as a negative regulator of thioredoxin. Here, we report the role of TBP-2 in oxidative renal tubular injury and the subsequent carcinogenesis by ferric nitrilotriacetate. TBP-2 was abundantly expressed in the rat kidney. Immunohistochemical analysis revealed that TBP-2 was present in association with nuclei and mitochondrial intermembrane space in the proximal tubular cells and coimmunoprecipitated with cytochrome c. After acute oxidative tubular damage, TBP-2 protein, but not messenger RNA, markedly decreased, demonstrating shortened half-life of this protein. Most cases of the induced renal cell carcinoma showed undetectable levels of TBP-2 protein, which was associated with the methylation of CpG island in the promoter region. Genome sequence analyses identified the poly-A tract in the 3' untranslated region as a mutation hot spot in this rather nonselective environment. Collectively, the amounts of TBP-2 protein were inversely associated with proliferation of tubular cells, as evaluated by proliferating cell nuclear antigen. These results suggest that loss of TBP-2 is essential for proliferation of not only neoplastic but also non-neoplastic renal tubular cells, and that TBP 2 is a target gene in oxidative stress-induced renal carcinogenesis by ferric nitrilotriacetate. PMID- 15834432 TI - CD52 and CD45 monoclonal antibodies for reduced intensity hemopoietic stem cell transplantation from HLA matched and one antigen mismatched unrelated donors. AB - Allogeneic hemopoietic stem cell transplantation (HSCT) is the only curative option for many patients with hematological malignancies. Since many of these patients lack HLA-identical sibling donors and are older or have comorbidity, a fully ablative HSCT is not feasible and an alternative approach is required. We studied 22 consecutive patients who could not have myeloablative conditioning because of comorbidity or age - 21/22 being over the age of 50 years (median 58 years range 20-70 years). A conditioning regimen consisting of fludarabine, total body radiation 450 cGy and alemtuzumab (CD52 mAb) was used for 15 patients. A second group of seven patients received CD45 monoclonal antibodies in addition. Unrelated donor stem cells were HLA matched (15 patients - 68%) or one locus mismatched (seven patients - 32%). In all, 16 patients had high-risk disease, including 12 with active malignancy at the time of transplant. With a median follow-up of 715 (216-1470) days, nonrelapse mortality, actuarial event-free and overall survival is 27, 45 and 45%, respectively. Hence the outcome of reduced intensity HSCT with lymphodepleting antibodies in older patients with intermediate/high-risk hematological malignancies appears comparable to that obtained with fully ablative transplantation in younger patients, even when these older recipients lack HLA-identical sibling donors. PMID- 15834434 TI - Acquired myeloma-associated Type III hyperlipidaemia treated by nonmyeloablative HLA-identical sibling allogeneic stem cell transplant using a donor with essential thrombocythaemia (ET): evidence of engraftment without manifestation of ET in recipient. PMID- 15834433 TI - Autologous and allogeneic stem cell transplantation in adult ALL: the Swedish Adult ALL Group experience. AB - Adult patients with acute lymphoblastic leukaemia (ALL) have been treated according to national protocols in Sweden since 1986. Stem cell transplantation (SCT) has been recommended in first remission for patients with risk factors for relapse, and for standard risk patients only after relapse. In this retrospective study, the results of autologous and allogeneic SCT in these populations were evaluated. In total, 187 patients with a median age of 34 years (17-66 years) underwent SCT. The 5-year disease-free survival (DFS), for all patients, was 26% (Confidence intervals (CI) 20-32%). The 5-year DFS was higher for patients transplanted in first remission 32% (CI 24-40%) compared to 14% (CI 5-23%; P<0.0001) in patients transplanted beyond first remission. No significant differences in DFS (P=0.06) were determined between autologous, related donor and unrelated donor SCT in the whole cohort. A lower relapse rate was counterbalanced by higher treatment-related mortality in patients undergoing allogeneic SCT. In Philadelphia-positive ALL, allogeneic SCT was superior to autologous SCT, with a 5-year DFS of 30% (CI 12-47%) vs 0% (P=0.04). Limited chronic graft-versus-host disease (GVHD) was associated with an improved DFS of 53% (CI 38-69%) compared to no chronic GVHD of 22% (CI 10-36%; P=0.0008), indicating a clinically important graft-versus-leukaemia effect. PMID- 15834435 TI - Results of autologous and allogeneic hematopoietic cell transplant therapy for multiple myeloma. AB - We compared the results of autologous and allogeneic peripheral blood hematopoietic cell transplant (HCT) in 87 patients with multiple myeloma using myeloablative preparative regimen. Autologous transplant (n=70) led to a lower 100-day transplant-related mortality (TRM) of 4% [0-9%] compared to 18% [0-36%] in allogeneic recipients (P=0.02). More frequent complete responses were seen in allogeneic recipients (64% [37-91%] vs 34% [23-45%] in autologous recipients, P=0.09). In autologous recipients, survival at 1 year was 86% [80-95%] and, it fell to 50% [47-75%] at 4 years, whereas in allogeneic recipients, survival at 1 and 4 years remained at 64% [40-87%]. In patients surviving more than one year, 4 year survival was superior in allogeneic (100%) vs autologous recipients (58% [41 75%], P=0.02). A trend toward higher relapse was seen in autologous transplant patients (73% [55-90%] vs 37% [11-63%] in allogeneic transplant patients, P=0.1). We observed good tolerance of myeloablative conditioning regimen followed by either autologous or allogeneic transplant. Although autologous HCT is associated with lower TRM, allogeneic HCT has acceptable TRM, and is more likely to provide a sustained response. Allogeneic HCT may be suitable in younger patients, soon after diagnosis, and in those with chemosensitive disease. PMID- 15834436 TI - What is the impact of HLA mismatches detected by high-resolution techniques on the outcome of unrelated donor bone marrow transplantation? PMID- 15834437 TI - The hemochromatosis C282Y allele: a risk factor for hepatic veno-occlusive disease after hematopoietic stem cell transplantation. AB - Hepatic veno-occlusive disease (HVOD) is a serious complication of hematopoietic stem cell transplantation (HSCT). Since the liver is a major site of iron deposition in HFE-associated hemochromatosis, and iron has oxidative toxicity, we hypothesized that HFE genotype might influence the risk of HVOD after myeloablative HSCT. We determined HFE genotypes in 166 HSCT recipients who were evaluated prospectively for HVOD. We also tested whether a common variant of the rate-limiting urea cycle enzyme, carbamyl-phosphate synthetase (CPS), previously observed to protect against HVOD in this cohort, modified the effect of HFE genotype. Risk of HVOD was significantly higher in carriers of at least one C282Y allele (RR=3.7, 95% CI 1.2-12.1) and increased progressively with C282Y allelic dose (RR=1.7, 95% CI 0.4-6.8 in heterozygotes; RR=8.6, 95% CI 1.5-48.5 in homozygotes). The CPS A allele, which encodes a more efficient urea cycle enzyme, reduced the risk of HVOD associated with HFE C282Y. We conclude that HFE C282Y is a risk factor for HVOD and that CPS polymorphisms may counteract its adverse effects. Knowledge of these genotypes and monitoring of iron stores may facilitate risk-stratification and testing of strategies to prevent HVOD, such as iron chelation and pharmacologic support of the urea cycle. PMID- 15834438 TI - The cardiovascular actions of anandamide: more targets? PMID- 15834439 TI - Two mechanisms of action of the adamantane derivative IEM-1460 at human AMPA-type glutamate receptors. AB - 1. Antagonizing glutamatergic neurotransmission by blockade of AMPA-type glutamate receptors (GluR) is a promising pharmacological strategy for neuroprotection in neurodegenerative diseases and acute treatment of stroke. 2. We investigated the interaction of the adamantane derivative IEM-1460 with human wild-type and mutant AMPA-type GluR channels. Different recombinant homooligomeric human AMPA-type GluR channels and a rat nondesensitizing mutant GluR (GluR2 L504Y) channel were expressed in HEK293 cells and investigated using the patch-clamp technique in combination with ultrafast agonist application. 3. When IEM-1460 was coapplied with glutamate, an open channel block mechanism was observed at slow desensitizing GluR2 flip (>/=0.1 mM IEM-1460) and nondesensitizing GluR2 L504Y channels (>/=1 microM IEM-1460). 4. A competitive block of AMPA-type channels was observed with IC(50) values for the dose block curves of 0.1 mM IEM-1460 at human unmutated and 10 microM IEM-1460 at mutant GluR channels. 5. Nondesensitizing GluR2 L504Y channels were used to further characterize the block mechanism. After equilibration with the agonist, a current decay upon coapplication of glutamate and IEM-1460 was observed. The recovery from block was independent of the glutamate and IEM-1460 concentration. The extent of current inhibition as well as the time constant of current decay upon addition of the blocker to the test solution were dependent on agonist concentration; this strongly points to an additional competitive-like block mechanism of IEM-1460 at human AMPA-type GluR channels. 6. The data were interpreted in the frame of a molecular scheme with two binding sites of IEM-1460 at the receptor, one at the unliganded resting and the other at the fully liganded open state of the channels. PMID- 15834440 TI - Increased alphaCGRP potency and CGRP-receptor antagonist affinity in isolated hypoxic porcine intramyocardial arteries. AB - 1. This study describes the effects of hypoxia on relaxing responses and cAMP production induced by the known vasodilator peptides: alphaCGRP, amylin (AMY) and adrenomedullin (AM) on isolated pig coronary arteries in vitro. 2. Hypoxic incubation increased the vasorelaxant effect of alphaCGRP (four-fold; P<0.05), AMY (3.2-fold; P<0.05), but not significantly for AM (two-fold; NS). 3. Whereas hypoxia had no effect on arterial cAMP levels, it significantly potentiated the production of cAMP stimulated of alphaCGRP and AMY, but not of AM. 4. The antagonist alphaCGRP(8-37) also exerted an increased effect in hypoxia. The Schild plot-derived pK(B) values revealed an increase in the apparent affinity of the antagonist for the CGRP(1) receptor from 7.0 to 7.2 under control conditions versus 8.0 in hypoxia. 5. Removal of endothelium, peptidase inhibitors, preincubation with the adenosine A(2A) receptor antagonist CSC (10(-3) M), the ATP-sensitive K-channel inhibitor glibenclamide (10(-5) M), the cyclooxygenase inhibitor indomethacin (10(-3) M) or NG-monomethyl-L-arginine (10(-4) M) had no effect on the alphaCGRP-induced vasorelaxation in hypoxia; neither did hypoxia influence the levels of CGRP and AM receptor mRNA. 6. We conclude that hypoxic incubation increases the relaxation and cAMP production induced by alphaCGRP and AMY in rings of porcine coronary arteries in vitro. A concomitant release of adenosine, a cyclooxygenase product, an endothelium-derived substance, activation of vascular ATP-sensitive K-channels, peptidase inhibitors or changes in CGRP and AM receptor mRNA cannot account for the changes observed in hypoxia. Moreover, alphaCGRP(8-37) showed increased affinity at the CGRP(1) receptor during hypoxia, possibly due to a conformational change at the CGRP(1) receptor site. PMID- 15834442 TI - Tramadol produces outward currents by activating mu-opioid receptors in adult rat substantia gelatinosa neurones. AB - 1. An action of a tramadol metabolite, mono-O-dimethyl-tramadol (M1), on substantia gelatinosa (SG) neurones in adult rat spinal cord slices was examined by using the whole-cell patch-clamp technique. 2. In 41% of the neurones examined, superfusing M1 produced an outward current at -70 mV; this response reversed at a potential close to the equilibrium potential for K(+). M1 current hardly declined and persisted for >30 min after its washout. 3. M1 current correlated in amplitude with current produced by mu-opioid receptor agonist DAMGO in the same neurone, and largely reduced in amplitude in the presence of mu opioid receptor antagonist CTAP but not alpha2-adrenoceptor antagonist yohimbine. In a neurone where M1 had no effect on holding currents, noradrenaline produced an outward current at -70 mV. 4. The amplitude of the M1 response, relative to that of the DAMGO response, exhibited an EC(50) value of 300 microM. 5. We conclude that M1 produces a persistent hyperpolarization by activating mu-opioid receptors in adult rat SG neurones. This could contribute to at least a part of pain alleviation produced by tramadol. PMID- 15834443 TI - Effect of galantamine on the human alpha7 neuronal nicotinic acetylcholine receptor, the Torpedo nicotinic acetylcholine receptor and spontaneous cholinergic synaptic activity. AB - 1. Various types of anticholinesterasic agents have been used to improve the daily activities of Alzheimer's disease patients. It was recently demonstrated that Galantamine, described as a molecule with anticholinesterasic properties, is also an allosteric enhancer of human alpha4beta2 neuronal nicotinic receptor activity. We explored its effect on the human alpha7 neuronal nicotinic acetylcholine receptor (nAChR) expressed in Xenopus oocytes. 2. Galantamine, at a concentration of 0.1 microM, increased the amplitude of acetylcholine (ACh) induced ion currents in the human alpha7 nAChR expressed in Xenopus oocytes, but caused inhibition at higher concentrations. The maximum effect of galantamine, an increase of 22% in the amplitude of ACh-induced currents, was observed at a concentration of 250 microM Ach. 3. The same enhancing effect was obtained in oocytes transplanted with Torpedo nicotinic acetylcholine receptor (AChR) isolated from the electric organ, but in this case the optimal concentration of galantamine was 1 microM. In this case, the maximum effect of galantamine, an increase of 35% in the amplitude of ACh-induced currents, occurred at a concentration of 50 microM ACh. 4. Galantamine affects not only the activity of post-synaptic receptors but also the activity of nerve terminals. At a concentration of 1 microM, quantal spontaneous events, recorded in a cholinergic synapse, increased their amplitude, an effect which was independent of the anticholinesterasic activity associated with this compound. The anticholinesterasic effect was recorded in preparations treated with a galantamine concentration of 10 microM. 5. In conclusion, our results show that galantamine enhances human alpha7 neuronal nicotinic ACh receptor activity. It also enhances muscular AChRs and the size of spontaneous cholinergic synaptic events. However, only a very narrow range of galantamine concentrations can be used for enhancing effects. PMID- 15834444 TI - Genomic and nongenomic effects of aldosterone in the rat heart: why is spironolactone cardioprotective? AB - 1. Mineralocorticoid receptor (MR) antagonism with spironolactone reduces mortality in heart failure on top of ACE inhibition. To investigate the underlying mechanism, we compared the actions of both aldosterone and spironolactone to those of angiotensin (Ang) II in the rat heart. 2. Hearts of male Wistar rats were perfused according to Langendorff. Ang II and aldosterone increased left ventricular pressure (LVP) by maximally 11+/-4 and 9+/-2%, and decreased coronary flow (CF) by maximally 36+/-7 and 20+/-4%, respectively. Spironolactone did not significantly affect LVP or CF. 3. In hearts that were exposed to a 45-min coronary artery occlusion and 3 h of reperfusion, a 15-min exposure to spironolactone prior to occlusion reduced infarct size (% of risk area) from 68+/-2 to 45+/-3%, similar to the reduction (34+/-2%) observed following 'preconditioning' (15 min occlusion followed by 10 min reperfusion) prior to the 45-min occlusion. Aldosterone exposure did not affect infarct size (71+/-5%). 4. In cardiomyocytes, aldosterone decreased [(3)H]thymidine incorporation maximally by 73+/-3%, whereas in cardiac fibroblasts it decreased [(3)H]proline incorporation by 33+/-7%. Spironolactone inhibited both effects. Ang II increased DNA and collagen synthesis, and these effects were reversed by aldosterone. 5. In conclusion, aldosterone induces positive inotropic and vasoconstrictor effects in a nongenomic manner, and these effects are comparable to those of Ang II. Aldosterone reduces DNA and collagen synthesis via MR activation, and counteracts the Ang II-induced increases in these parameters. MR blockade reduces infarct size and increases LVP recovery following coronary artery occlusion. The MR-related phenomena may underlie, at least in part, the beneficial actions of spironolactone in heart failure. PMID- 15834445 TI - Central and peripheral components of the pressor effect of anandamide in urethane anaesthetized rats. AB - 1. We wanted to search for the mechanism(s) responsible for the brief pressor response induced by anandamide in urethane-anaesthetized rats. 2. The anandamide induced pressor effect was not modified by the antagonists of cannabinoid CB(1) and vanilloid TRPV(1) receptors, SR 141716A (3 micromol kg(-1)) and capsazepine (1 micromol kg(-1)), respectively, by bilateral vagotomy and by pithing. Replacement of urethane by pentobarbitone virtually abolished the pressor effect of anandamide, both in pithed and vagotomized and in 'intact' rats (i.e. not treated in this manner). 3. The pressor effect of anandamide was reduced by the nonselective TRPV family inhibitor ruthenium red (3 micromol kg(-1)) and by the blocker of L-type calcium channels nifedipine (1 micromol kg(-1)), both in pithed urethane-anaesthetized rats and in 'intact' urethane-anaesthetized rats. The nonselective beta-adrenoceptor antagonist propranolol (0.1 or 0.3 micromol kg( 1)) and the nonselective NMDA receptor antagonist MK-801 (1 micromol kg(-1)) diminished the anandamide-induced vasopressor response in 'intact' but not in pithed rats. The inhibitory effect of propranolol in 'intact' rats was mimicked by the beta(2)-adrenoceptor antagonist ICI 118551 (1 micromol kg(-1)), but not by the beta(1)-adrenoceptor antagonist CGP 20712 (1 micromol kg(-1)). 4. The present study revealed that two mechanisms may be responsible for the anandamide-induced pressor response in urethane-anaesthetized rats. The first involves the central nervous system (probably the medulla oblongata) and is sensitive to propranolol and MK-801. The second, which is located peripherally (most probably in blood vessels), is sensitive to nifedipine, ruthenium red and pentobarbitone and, hence, probably represents a Ca(2+)-dependent mode of action. PMID- 15834446 TI - Effects of amantadine and budipine on antidepressant drug-evoked changes in extracellular 5-HT in the frontal cortex of freely moving rats. AB - 1. Evidence has recently suggested that NMDA receptors may play a role in the aetiology and possible treatment of depression and that weak noncompetitive NMDA receptor antagonists such as amantadine can synergize with conventional antidepressants in a model of the illness. 2. To try to obtain a neurochemical rationale for these findings, we have studied the effects of acute and chronic administration of amantadine or the related drug budipine on cortical release of 5-hydroxytryptamine (5-HT) following the antidepressants reboxitine (REB), paroxetine (PAROX) and clomipramine (CLOM) in freely moving rats by using microdialysis. 3. Acute administration of amantadine (40 mg kg(-1)), budipine (10 mg kg(-1)), REB (10 mg kg(-1)), PAROX (10 mg kg(-1)) or CLOM (10 mg kg(-1)) all failed to significantly alter extracellular 5-HT in the cortex. However, when either amantadine or budipine was administered 30 min prior to any of the three antidepressants, a significant rise in 5-HT was observed. 4. For chronic studies, the effects of the drugs were studied at 4, 7, 14 and 21 days. Amantadine and budipine did not significantly alter extracellular 5-HT at any time point. The three antidepressant drugs all elicited a gradual increase in 5-HT, which became significant after 14 days and tended to plateau thereafter. When either amantadine (20 mg kg(-1)) or budipine (5 mg kg(-1)) was coadministered with any of the three antidepressants, two differences were seen compared with the effects of the antidepressants alone. Firstly, the time required for significant increases in cortical 5-HT was reduced with elevated levels now being observed by 7 days. Secondly, the absolute magnitude of the increase in extracellular 5-HT was markedly greater in these rats from day 7 until the end of the experiment. 5. If, as is widely considered, an increase in extracellular 5-HT represents a critical step in the mechanism of action of antidepressants, these data suggest that combined treatment with clinically tolerated NMDA antagonists such as amantadine could reduce the delay in therapeutic onset of antidepressants as well as possibly enhance their efficacy. PMID- 15834447 TI - Actions of tacrine and galanthamine on histamine-N-methyltransferase. AB - Histamine-synthesizing neurons in the brain may play an important role in cognition, and a histaminergic deficit has been found in Alzheimer's disease (AD). The AD medication tacrine was previously shown to inhibit some forms of rodent histamine-N-methyltransferase (HNMT), but the effects of AD drugs have not been investigated on human HNMT activity. Presently, the effects of tacrine and galanthamine (another AD medication) were studied on the activity of several forms of human and rat HNMT. Tacrine (0.01-10 microM) inhibited both human and rat HNMT activity in a concentration-dependent manner, but was less potent on both human embryonic kidney and recombinant human brain HNMT than on rat kidney HNMT (IC50 values were 0.46 and 0.70 microM vs. 0.29 microM, respectively). Galanthamine (up to 10 microM) did not influence the activity of rat kidney or human HNMT. Tacrine, but not galanthamine, may achieve brain levels sufficient to influence histamine metabolism in some patients treated for AD. PMID- 15834448 TI - The effect of beta-carotene on the photostability of nisoldipine. AB - The effect of beta-carotene on the photostability of nisoldipine (NS) has been investigated. Light stability studies were carried out following the recommendations of the International Conference on Harmonization (ICH), version I, using a high-pressure mercury lamp, type HBO-50, equipped with the interference filter and Wood's filter to isolate 365 nm wavelength. The photodegradation process was assayed by means of UV spectrophotometry and the reversed phase High-Performance Liquid Chromatography (HPLC). The quantum yields of photodegradation of NS and NS in the presence of beta-carotene were determined using the Reinecke salt as a chemical actinometer. The methanol solutions of NS degraded following apparent first-order kinetics. The degradation rate constant decreased as beta-carotene concentration increased. To verify the photostabilising role of the dye, a Stern-Volmer plot was constructed. The results have proved that beta-carotene does not act as a photosensitiser. PMID- 15834450 TI - Genistein inhibited hypoxia-inducible factor-1alpha expression induced by hypoxia and cobalt chloride in human retinal pigment epithelium cells. AB - The time-dependent changes of hypoxia-inducible factor-1alpha (HIF-1alpha) expression induced by hypoxia and CoCl2 treatment and the effects of genistein on the level of HIF-1alpha expression in human retinal pigment epithelium cells were examined. Judging by relative fluorescence using a confocal scanning laser microscope coupled to a computer, HIF-1alpha expression was determined. It was found that hypoxia could markedly increase the expression of HIF-1alpha. The highest expression of HIF-1alpha was detected at 1 h, which was 313.9% +/- 38.2% of the control level. After pretreatment with genistein (50, 100, and 200 micromol/l), the hypoxia-evoked HIF-1alpha expression was concentration dependently inhibited. CoCl2 treatment could significantly elevate the level of HIF-1alpha expression. At 0.5 h after CoCl2 treatment, the highest level was observed, which was 141.4% +/- 14.1% of the control level. Genistein 50, 100, 200 micromol/L could also suppress HIF-1alpha expression in a concentration-dependent manner. These results suggested that the inhibition of HIF-1alpha protein expression by genistein may partly account for its effect on retinal neovascularization in vivo. PMID- 15834449 TI - Association of cigarette smoking with steady-state plasma concentration of irbesartan in male Chinese with hypertension. AB - Previous studies suggested that cigarette smoking, being highly prevalent in many countries, is an important environmental factor that contributes to interindividual variations in response to certain medications. To investigate the possible interactions between irbesartan (a commonly used angiotensin II type 1 receptor antagonist) and cigarette smoking, we recruited 491 male patients with essential hypertension from two rural districts of China. All subjects were treated with irbesartan (150 mg/day) for 28 days, and the steady-state plasma concentrations of irbesartan 24 h after the dose of day 27 (the trough level) and 6 h after the dose of day 28 were determined using prevalidated high-performance liquid chromatography (HPLC)-fluorescence method. Multiple linear regression was used to analyze the effect of smoking on the steady-state plasma concentrations of irbesartan, with potential confounding factors adjusted. The results showed that current smokers had significantly higher average steady-state trough plasma level of irbesartan than that of the nonsmokers and former smokers. The finding was still valid when subjects from the two different districts were separately analyzed. The stratified analysis according to age suggested that the effect of increasing irbesartan concentration by current cigarette smoking in the elderly patients was stronger than that in adults. The metabolism of irbesartan in current smokers was slower compared to that in nonsmokers and former smokers. In summary, the present study showed significant associations between current cigarette smoking and an increased steady-state trough plasma concentration of irbesartan in a male Chinese population with hypertension. The metabolic rate of irbesartan in current smokers was slower than those in nonsmokers and former smokers. PMID- 15834451 TI - Death by stress: natural IgM-induced apoptosis. AB - Stress kills and hence should be avoided. On the other hand, stress induction can be used to remove malignant cells by inducing cellular suicide. Natural IgM antibodies act as first-line defense in immune surveillance. These antibodies selectively kill aberrant cells by using different apoptotic stress mechanisms. They can be isolated from patients but also from healthy donors by using the human hybridoma technology. They are components of the innate immunity, and, on the basis of specific screening methods, should also be detectable in any other individual. The three tumor-specific, apoptosis-inducing natural IgM antibodies described in this review are good examples for stress-induced apoptosis and nature's resourceful ways to fight malignant growth. PMID- 15834452 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity. prous.com. This issue focuses on the following selection of drugs: ABX-IL-8, Acclaim, adalimumab, AGI 1067, alagebrium chloride, alemtuzumab, Alequel, Androgel, anti-IL-12 MAb, AOD 9604, aripiprazole, atomoxetine hydrochloride; Biphasic insulin aspart, bosentan, botulinum toxin type B, bovine lactoferrin, brivudine; Cantuzumab mertansine, CB 1954, CDB-4124, CEA-TRICOM, choriogonadotropin alfa, cilansetron, CpG-10101, CpG 7909, CTL-102, CTL-102/CB-1954; DAC:GRF, darbepoetin alfa, davanat-1, decitabine, del-1 Genemedicine, dexanabinol, dextofisopam, dnaJP1, dronedarone hydrochloride, dutasteride; Ecogramostim, eletriptan, emtricitabine, EPI-hNE-4, eplerenone, eplivanserin fumarate, erlotinib hydrochloride, ertapenem sodium, escitalopram oxalate, esomeprazole magnesium, etoricoxib, ezetimibe; Falecalcitriol, fingolimod hydrochloride; Gepirone hydrochloride; HBV-ISS, HSV-2 theracine, human insulin; Imatinib mesylate, Indiplon, insulin glargine, ISAtx-247; L612 HuMAb, levodopa/carbidopa/entacapone, lidocaine/prilocaine, LL-2113AD, lucinactant, LY 156735; Meclinertant, metelimumab, morphine hydrochloride, morphine-6 glucuronide; Natalizumab, nimotuzumab, NX-1207, NYVAC-HIV C; Omalizumab, onercept, osanetant; PABA, palosuran sulfate, parathyroid hormone (human recombinant), parecoxib sodium, PBI-1402, PCK-3145, peginterferon alfa-2a, peginterferon alfa-2b, peginterferon alfa-2b/ribavirin, pemetrexed disodium, pimecrolimus, PINC, pregabalin; Ramelteon, rasagiline mesilate, rasburicase, rimonabant hydrochloride, RO-0098557, rofecoxib, rosiglitazone maleate/metformin hydrochloride; Safinamide mesilate, SHL-749, sitaxsentan sodium, sparfosic acid, SprayGel, squalamine, St. John's Wort extract, synthetic human secretin; Taxus, telavancin hydrochloride, telithromycin, temoporfin, tenofovir disoproxil fumarate, tenofovir disoproxil fumarate/emtricitabine, teriparatide, testosterone gel, TG-1024, tirapazamine, travoprost, travoprost/timolol; Valdecoxib, valganciclovir hydrochloride, voriconazole; Ximelagatran. PMID- 15834453 TI - Effects of potassium adaptation on blood pressure and pressor responses in normotensive and renal hypertensive Wistar rats. AB - Potassium adaptation reduces blood pressure (BP) in hypertensive humans and animals but its effects on normotensive BP and the nature of pressor responses to vasoactive drugs are not known. We measured directly, the mean arterial pressure (MAP) of normotensive control, normotensive potassium-adapted (given 0.75% potassium chloride solution for 5 weeks), renal hypertensive (RHP), and renal hypertensive Wistar rats later adapted to potassium. The maximum percentage change, the ED25, and recovery times after bolus injections of noradrenaline (NA), angiotensin II (Ang. II), sodium nitroprusside (SNP), and acetylcholine (ACh) were compared. The MAP of normotensive potassium-adapted rats was significantly lower than that of the normotensive controls (95.6+/-5.0 vs. 110.8+/-2.8 mmHg, p<0.05). The potassium-adapted hypertensive rats (RHP-A) also had significantly lower MAP values than the non-adapted hypertensive ones (116.0+/-4.4 vs. 138.2+/-4.1 mmHg, p<0.01). Potassium adaptation significantly blunted responses to NA and augmented responses to SNP but while the duration of action of Ang. II was significantly shortened, that of SNP was significantly increased. We conclude that potassium adaptation reduces BP in the normotensive and hypertensive rats and may influence both the degree and duration of action of vasoactive drugs given as bolus injections. PMID- 15834454 TI - Effect of rofecoxib on antihypertensive effects of candesartan in experimental models of hypertension. AB - Given the high prevalence of hypertension, concomitant use of nonsteroidal anti inflammatory drugs and antihypertensive medications is commonly encountered in clinical practice. The present study was designed to study the effect of indomethacin, nimesulide, and rofecoxib on blood pressure (BP) in normotensive and hypertensive rats and also to investigate the effect of rofecoxib on BP control in candesartan-treated hypertensive rats. Male Wistar rats weighing 150 200 g were divided into three groups: control, DOCA-hypertensive, and L-NAME hypertensive rats. All the rats were given indomethacin (15 mg/kg body weight), nimesulide (20 mg/kg body weight), rofecoxib (10 mg/kg body weight), or vehicle orally and daily for 6 weeks. Hypertensive rats in separate groups were treated with either candesartan (1 mg/kg body weight) alone or a combination of candesartan (1 mg/kg body weight) and rofecoxib (10 mg/kg body weight) orally and daily for 6 weeks. BP measurements were performed using tail cuff method at baseline and 1-week intervals throughout the treatment period. All the three COX inhibitors resulted in increase in BP, but mean change in BP was the highest with rofecoxib. Rofecoxib-treated L-NAME-hypertensive rats exhibited a significant increase in mean arterial pressure at 6 weeks (168.3+/-5.7 mmHg) as compared with DOCA-hypertensive rats (128.818+/-7.2 mmHg). Administration of Rofecoxib L-NAME hypertensive rats treated with candesartan resulted in a significant increase in BP. Systolic BP at 0 week (107.0+/-4.2 mmHg) rose to 141.6+/-2.0 mmHg at 6 weeks. Systolic BP at 2, 4, and 6 weeks was significantly higher as compared with (L NAME+candesartan)- and (rofecoxib+candesartan)-treated group. In conclusion, concomitant use of rofecoxib resulted in poor BP control by candesartan in L-NAME hypertensive rats. PMID- 15834455 TI - The effect of nicotinic acid and alcohol co-administration in Wistar rats. AB - The effects of co-administration of nicotinic acid (NA) and alcohol (Alc) on liver function in male Wistar rats were evaluated. The rats were randomized into five groups: (i) Olive oil (Oil), (ii) Alc+Oil, (iii) NA+Oil, (iv) NA+Alc+Oil, and (v) Controls (fed only normal rat chow). Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AP), total cholesterol and triglycerides (TG) were measured. Liver histopathology was also assessed. The Alc+Oil group had higher TG levels compared with the NA+Alc+Oil group and all other groups, as well. NA+Oil group had higher levels of AP compared with Alc+Oil and Oil groups. The NA+Oil group had higher ALT levels compared with the Oil group. The Oil group had lower ALT levels compared with the control group. The Alc+Oil group had higher AST levels compared with the NA+Alc+Oil group, as well as with all other groups. Liver histopathology was within the normal range. A moderate amount of Alc daily together with NA is safe in rats. The NA and Alc co administration reduces the TG and AST levels in rats, compared with the administration of Alc alone. PMID- 15834456 TI - Pharmacodynamic effects of 3-day intravenous treatment with pantoprazole or ranitidine after 10 days of oral ranitidine. AB - Tachyphylaxis (drug tolerance) is an undesirable condition in drug therapy with histamine-2-receptor antagonists (H2RAs). The concept of overcoming tachyphylaxsis via intravenous (i.v.) administration of proton-pump inhibitors (PPIs) or H2RAs is of significant interest to physicians. In the present study, 32 healthy Helicobacter pylori negative male volunteers were evaluated for the ability of i.v. pantoprazole or i.v. ranitidine to overcome oral ranitidine tachyphylaxis. After 10 days of oral treatment with enteric-coated 300-mg ranitidine tablets once daily in the evening, two groups of 16 volunteers each were randomized to receive either i.v. pantoprazole or i.v. ranitidine for up to 72 h. The primary variable was defined as the increase in 24-h gastric pH median after 1 day of i.v. treatment; the secondary variable was median percentage of time that 24-h gastric pH was <4, as calculated by Hodges-Lehman shift estimators. After 10 days of oral ranitidine treatment, tachyphylaxis was present in all volunteers. Within 1 day of continuous i.v. pantoprazole or i.v. ranitidine administration, 24-h median gastric pH increased from pH 1.45 to pH 3.50 (241%) and from pH 1.50 to pH 2.35 (157%), respectively. I.v. pantoprazole was found to be significantly more effective (p<0.05) than i.v. ranitidine in increasing the 24-h gastric pH after oral ranitidine tachyphylaxis. PMID- 15834457 TI - Effects of NMDA and MK-801 injected into the substantia nigra pars reticulata on jaw movements evoked by dopamine D1-/D2 receptor stimulation in the ventrolateral striatum: studies in freely moving rats. AB - The effects of NMDA and MK-801 injected into the substantia nigra pars reticulata on jaw movements evoked by dopamine D1/D2 receptor stimulation in the ventrolateral striatum were examined in freely moving rats, by using a magnet sensing system combined with intracerebral drug microinjection technique. Bilateral injections of a mixture of SKF 82958 (5 microg) and quinpirole (10 microg), agonist at dopamine D1 and D2 receptors respectively, into the ventrolateral striatum elicited repetitive jaw movements. Bilateral injections of NMDA (0.01 and 0.05 microg/0.2 microl in each side) into the substantia nigra pars reticulata, which alone did not produce jaw movements, reduced the repetitive jaw movements evoked by the dopamine D1/D2 receptor agonist mixture in a dose-dependent manner. Injection of the non-competitive NMDA receptor antagonist, MK-801 (0.1 and 0.5 microg/0.2 microl in each side), into the substantia nigra pars reticulata, which alone did not produce jaw movements, prevented the dopaminergic jaw movements in a dose-dependent manner. Moreover, other behaviors such as grooming, rearing, yawning, vacuous chewing, and locomotor activity that occurred after injections of the dopamine receptor agonist mixture were not significantly altered by the bilateral injections of NMDA or MK-801 into the substantia nigra pars reticulata. Given our previous results showing that both agonist and antagonist of GABA(A) receptors injected into the substantia nigra pars reticulata inhibit the jaw movements elicited by dopamine D1/D2 receptor stimulation in the ventrolateral striatum, the present results suggest that there are complex functional interactions between NMDA and GABA(A) receptors within the substantia nigra pars reticulata that may be responsible for the common profiles in the effects of NMDA and GABA(A) receptor agents. PMID- 15834458 TI - Rhabdomyolysis. AB - Rhabdomyolysis is a condition caused by skeletal muscle injury and release of muscle cell contents into the circulation. It may result in myoglobinuria, the filtration of myoglobin into the urine, and is often associated with acute renal failure (ARF). Rhabdomyolysis may complicate many disease states. In some, such as crush injury, muscle injury is obvious; in others, such as drug overdose, it may never be apparent. It may occur in the setting of an altered mental status, and even in the conscious patient, it may occur with minimal symptoms or physical findings. Therefore, diagnosis requires a high level of suspicion and appropriate sensitivity to abnormal laboratory values. Many insults can precipitate rhabdomyolysis and myoglobinuria. Disruption of the muscle cell membrane may result from a direct mechanical or toxic insult to the membrane or an inability to maintain ionic gradients across the membrane (as in ischemia or extreme exertion). This article reviews the etiology, pathogenesis, clinical features, complications, and management of rhabdomyolysis, particularly crush injuries in the setting of a major disaster. PMID- 15834459 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials reported in current literature and congresses. The data in the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs:[188Re]-HDD; A-179578, adalimumab, AK-602, albumin interferon alfa, alfimeprase, amelubant, anakinra, anti-CD2 MAb, APD-356, aripiprazole, atvogen; Bimatoprost, bimosiamose, BLP-25, brivaracetam; Caspofungin acetate, cilansetron, CMV vaccine (bivalent), conivaptan hydrochloride, Cypher; Darbepoetin alfa, darifenacin hydrobromide, D D4FC, decitabine, dnaJP1, doranidazole, dronedarone hydrochloride; Efalizumab, efaproxiral sodium, emtricitabine, Endeavor, entecavir, erlotinib hydrochloride, escitalopram oxalate, etoricoxib, etravirine, ezetimibe; Fampridine, fenretinide, ferumoxtran-10, forodesine hydrochloride; Gantacurium chloride, gemi-floxacin mesilate, Glyminox, GW-501516; HBV-ISS, hepavir B, human insulin, HuMax-CD20, hyaluronic acid, HyCAMP; Icatibant, IDEA-070, IGN-311, imatinib mesylate, insulin detemir, insulin glargine, insulin glulisine; Lapatinib, lasofoxifene tartrate, LB-80380, liarozole fumarate, liposome encapsulated doxorubicin, lumiracoxib, LY 570310; MC-1, melatonin, merimepodib, metanicotine, midostaurin; Natalizumab, nicotine conjugate vaccine, NYVAC-HIV C; Patupilone, peginterferon alfa-2a, peginterferon alfa-2b, peginterferon alfa-2b/ribavirin, pelitinib, Peru-15, pexelizumab, PHP, pimecrolimus, prednisolone sodium metasulfobenzoate; Recombinant alfa1-antitrypsin (AAT), retigabine, rHA influenza vaccine, rifalazil, rofecoxib, rosiglitazone maleate/Metformin hydrochloride, rostaporfin, rosuvastatin calcium, rubitecan; Selenite sodium, semilente insulin, SMP-797, sorafenib; Talampanel, tenofovir disoproxil fumarate, TER-199, tiotropium bromide, torcetrapib, treprostinil sodium, TTA; ValboroPro, valdecoxib, val-mCyd, valtorcitabine dihydrochloride: XP-828L. PMID- 15834460 TI - Metabolic mechanism of quetiapine in vivo with clinical therapeutic dose. AB - The in vivo metabolic mechanism of quetiapine (QTP) with clinical therapeutic dose was studied. Nineteen patients received multiple doses of QTP with or without concomitant erythromycin. Midazolam was given to detect enzyme activity. Plasma Concentrations of QTP, midazolam, and their metabolites were measured at specified time intervals. In presence of erythromycin, activity of CYP3A4 decreased significantly; for QTP, C(max), AUC(0-24), and t(1/2) increased significantly, CL decreased significantly, and variations in AUC(0-24) and CL showed, respectively, significant negative and positive correlation to that of CYP3A4 activity; for QTP sulfoxide (QTP-SF), C(max) and AUC(0-24) decreased significantly, t(1/2) increased significantly, and variation of t(1/2) was significantly positively correlated to that of CYP3A4 activity; for 7-hydroxy quetiapine (QTP-H), t(1/2) increased significantly and was closely correlated to CYP3A4 activity; for 7-hydroxy-N-desalkyl-quetiapine (QTP-ND), C(max) and AUC(0 24) decreased significantly, and variation of AUC(0-24) was significantly positively correlated to that of CYP3A4 activity. In conclusion, the major metabolic pathway of QTP is sulfoxidation. CYP3A4 is the primary enzyme responsible for CYP-mediated metabolism of QTP in clinical therapy dosage in vivo. QTP sulfoxidation and N-dealkylation are mainly catalyzed by CYP3A4. 7 Hydroxylation of QTP is not mainly catalyzed by CYP3A4. The metabolism of QTP-SF and QTP-H is mainly catalyzed by CYP3A4, but QTP-ND is not by CYP3A4. PMID- 15834461 TI - Ocular pharmacology of bicyclic hexahydroaporphines. AB - This paper explores the ocular hypotensive actions of bicyclic analogs of hexahydroaporphine (HHA), specifically nor-HHA, in an attempt to shed light on the mechanism(s) by which they lower intraocular pressure (IOP). Studies involving the measurement of IOP and aqueous humor production were conducted in ocular normotensive albino rabbits, while those involving smooth muscle contractility utilized isolated bovine iris. The ability of nor-HHA to produce a sustained drop in IOP is linked to both a functioning adrenergic nervous system and the availability of the products of cyclooxygenase metabolism. Although aqueous flow is not impacted by the bicyclic structures, the significant enhancement of outflow facility points to a probable mechanism of IOP-lowering action. Nor-HHA had no direct contractile or relaxant action on bovine irides, but does cause a concentration-dependent inhibition of carbachol-evoked contractions. This inhibition was reversed by inhibitors of phospholipase A(2) and cyclooxygenase, but not by inhibitors of lipoxygenase, again indicating a role for prostaglandins in the ocular pharmacological action of bicyclic HHAs. Pretreatment with a nitric oxide (NO) scavenger also reversed the ability of nor HHA to inhibit carbachol-induced contractions, implying a role for NO in the postjunctional actions of HHAs. PMID- 15834462 TI - Criticality of pH for accurate fluorometric measurements of dipyridamole levels in biological fluids. AB - Extended release dipyridamole (DIP) is widely used in clinical practice as an Aggrenox formulation, which is proven to improve outcomes for secondary stroke prevention in patients after acute vascular events. However, presently established fluorometry techniques are not suitable for trace amount determinations, because of the variable background fluorescence. The authors sought to determine whether biological fluid pH is important for the serial measures of DIP levels in the animal experiments and in patients treated with Aggrenox after ischemic stroke. Post-stroke patient (n = 34) and mice (n = 25) samples were tested to determine DIP levels by established techniques with FluoroMax 3 spectrofluorometer. Both the absorption and emission spectra of DIP were affected by modifications in pH. Fluorescence of DIP was found to be maximal at a wavelength of 490 nm (excitation 420 nm) and the spectral pattern was independent of pH. The intensity of fluorescence, however, was drastically lower at low pH (at pH 2.6, fluorescence was 4% of intensity at pH 9.8). Background plasma fluorescence, however, was completely unaffected by changes in pH. Using these fluorometric characteristics, a regression model that facilitates the efficient and sensitive determination of DIP concentration in biological fluids was formulated. Exploiting pH-dependent characteristics of DIP versus serum fluorescence patterns permits a convenient mathematical model to determine DIP concentration. This relatively inexpensive and time-efficient procedure can quantify drug levels in human/animal plasma/serum, thereby directly determining the level of patient adherence to the prescribed drug regimen, be it in the context of clinical trials or compliance with the animal protocol. PMID- 15834463 TI - An update on cyclic nucleotide phosphodiesterase (PDE) inhibitors: phosphodiesterases and drug selectivity. AB - In recent years, there has been a prodigious expansion of knowledge about cyclic nucleotide phosphodiesterases (PDEs). This isozyme superfamily has now become a major focus of drug discovery efforts owing to its diversity, molecular nature, differential regulation and expression in different cell types, and the range of biological functions. Inhibition of these isozymes can lead to an increase in cAMP and cGMP levels, which can affect a variety of physiological responses. Selective inhibitors for each of the multiple forms of PDE can offer an opportunity for desired therapeutic intervention and would be an extremely useful tool in drug discovery efforts for a medicinal chemist. This review details many key aspects of multiple forms of PDEs and their inhibitors with diversified chemical structures, which can act as leads for synthesis of novel drugs. PMID- 15834465 TI - Solid lipid nanoparticles as drug delivery systems. AB - For a decade, trials have been made to utilize solid lipid nanoparticles (SLNs) as alternative drug delivery systems to colloidal drug delivery systems such as lipid emulsions, liposomes, and polymeric nanoparticles. Various lipid matrices, surfactants, and other excipients used in formulation, preparation methods, sterilization and lyophilization of SLNs are discussed in this article. Entrapment efficiency of drug carrier and its effect on physical parameters, drug release, and release mechanisms of various compositions are reviewed and discussed. Important points in characterization and stability of SLNs are outlined. Various in vitro studies carried out by different research groups are mentioned in addition to in vivo evaluation. Exploitation potential of SLNs to administer by various routes of administration are covered. Passive and active drug targeting using SLNs are presented. PMID- 15834464 TI - Clinical evidence for drug treatments in obesity-associated hypertensive patients -a discussion paper. AB - The association between obesity and hypertension is well known. The hemodynamic features of obesity-related hypertension are an expansion of extracellular volume inducing hypervolaemia and increased cardiac output, with activation of both the sympathetic nervous system and the renin--angiotensin system. It is suggested that obesity-related hypertension may be considered as a subset of essential hypertension, and treated as an identity. Orlistat and sibutramine both reduce body weight in the obese patients. The use of orlistat in obese hypertensive patients is associated with a small decrease in blood pressure, whereas sibutramine may increase the blood pressure. Thus, orlistat may be preferred in the obese hypertensive patients. Diuretics and beta-blockers decrease insulin sensitivity, which is an unwanted effect in obesity, and should be used with caution in obese hypertensive patients. The calcium channel blockers have no or minor effects on insulin sensitivity and may be considered for use in obese hypertensive patients. Inhibitors of the effects of angiotensin may be the antihypertensive drugs of choice for obese hypertensive patients, as in addition to reducing blood pressure, ACE inhibitors and AT(1) receptor antagonists have no effect or improve insulin sensitivity, and are renoprotective. More clinical trials are needed for the centrally acting antihypertensives (clonidine, rilmenidine) in obese hypertensive patients, as they inhibit the sympathetic nervous and renin--angiotensin systems, which are overactive in this population. PMID- 15834466 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: 3-AP, 667-coumate, 9 aminocamptothecin; Ad5CMV-p53, AES-14, alefacept, anecortave acetate, APC-8024, APD-356, asoprisnil; Bevacizumab, bimakalim, bimatoprost, BLP-25, BR-1; Caspofungin acetate, cetuximab, cypher; Darbepoetin alfa, dexanabinol, dextromethorphan/quinidine sulfate, DNA.HIVA; Efaproxiral sodium, ertapenem sodium; Frovatriptan; HuMax-EGFr, HYB-2055, gamma-hydroxybutyrate sodium, Id-KLH vaccine, imatinib mesylate; Lapatinib, lonafarnib, Motexafin lutetium, MVA.HIVA, mycophenolic acid sodium salt; Nesiritide, NS-2330; Olmesartan medoxomil; Peginterferon alfa-2a, peginterferon alfa-2b, peginterferon alfa-2b/ribavirin, pemetrexed disodium, perifosine, pimecrolimus, pregabalin; QbG-10; Ralfinamide, rasburicase, rFGF-2, Ro-31-7453; Sitaxsentan sodium, sorafenib; Tadalafil, TC 1734, telmisartan/hydrochlorothiazide, tenofovir disoproxil fumarate, thymus nuclear protein, tipifarnib; Vandetanib, vibriolysin, vildagliptin, voriconazole. PMID- 15834467 TI - Functionalized base-pairs: versatile scaffolds for self-assembly. AB - This article discusses the development of synthetic supramolecular systems derived from hydrogen bond driven base-pairing, with a focus on the self-assembly of individual nucleobase analogues. PMID- 15834468 TI - Direct, trace level detection of explosives on ambient surfaces by desorption electrospray ionization mass spectrometry. AB - Desorption electrospray ionization (DESI) mass spectrometry is used to detect trace amounts of explosives present on a variety of ambient surfaces in 5-second analysis times without any sample preparation. PMID- 15834469 TI - Detection of explosives on solid surfaces by thermal desorption and ambient ion/molecule reactions. AB - A simple, fast and direct method is presented for detecting traces of solid explosives on cotton swabs or in particulate samples: ions are transferred into a mass spectrometer after thermal desorption and corona discharge chemical ionization in ambient air; specificity is enhanced using ambient ion/molecule reactions or by conventional tandem mass spectrometry. PMID- 15834470 TI - End-functionalization of poly(3-hydroxybutyrate)via genetic engineering for solid surface modification. AB - A new approach to end-functionalization of poly(3-hydroxybutyrate)[PHB] is described. Using genetically engineered PHB synthase fused with a 10x-histidine units at its N-terminus, end-functionalized PHB was synthesized and used for the solid surface modification. PMID- 15834471 TI - Expanding diversity in dynamic combinatorial libraries: simultaneous exchange of disulfide and thioester linkages. AB - Dynamic combinatorial libraries have been prepared which feature two simultaneous covalent exchange reactions in aqueous solution at neutral pH. This allows for diversity, not only of the subunits that are linked, but also of the linkage itself. PMID- 15834472 TI - 'Molecular capsules' based on p-sulfonatocalix[6]arene shrouding two tetraphenylphosphonium cations. AB - p-Sulfonatocalix[6]arene in the double cone conformation forms a molecular capsule-like arrangement confining two tetraphenylphosphonium cations, as part of an extended structure involving layers of additional tetraphenylphosphonium cations, aquated lanthanide ions and a large array of water molecules. PMID- 15834473 TI - The synthesis, characterisation and electronic structure of N-heterocyclic carbene adducts of P(I) cations. AB - Two different syntheses of N-heterocyclic carbene complexes of P(I) cations are presented; the structural features of these phosphamethine cyanine salts, in conjunction with the results of computational investigations, provide insight into the nature of the bonding of these heavy allene analogues. PMID- 15834474 TI - Amphiphilic behavior of an apparently non-polar calixarene. AB - The self-assembly properties of a non-polar calixarene have been investigated at the air-water interface, in water and in the solid state showing that this molecule behaves as an amphiphilic molecule. PMID- 15834475 TI - Synthesis of gold nanoparticles within a supramolecular gel-phase network. AB - Gold nanoparticles with diameters of ca. 13 nm were synthesised by UV irradiation of a supramolecular organogel into which HAuCl4 and tetraoctylammonium bromide had been diffused-the gel network plays an essential role in nanoparticle stabilisation. PMID- 15834476 TI - Solution-processed, highly-oriented supramolecular architectures of functionalized porphyrins with extended electronic states. AB - Thin films of aligned supramolecular architectures built from newly synthesized thiophene-substituted porphyrins have been processed from solution on surfaces. PMID- 15834477 TI - Adsorbate conformation determines catalytic chemoselectivity: crotonaldehyde on the Pt(111) surface. AB - Molecular orientation, which depends on surface coverage, determines whether or not catalytic hydrogenation is chemoselective. PMID- 15834478 TI - Fabrication of quantum dot-lectin conjugates as novel fluorescent probes for microscopic and flow cytometric identification of leukemia cells from normal lymphocytes. AB - The present study describes a synthesis of QD-lectin conjugates and their application for identification of leukaemia cells from normal lymphocytes using fluorescent confocal microscopy and flow cytometry. The results are compared with commercially available FITC-lectin. PMID- 15834479 TI - A 2D honeycomb-shaped network based on a starburst cluster: [Ag4(micro3 Cl)(PPh2(CH2)2PPh2)1.5{S2P(OR)2}3](R = Et, Pri). AB - The first 2D metal-organic framework utilizing PPh2(CH2)2PPh2 (dppe) units as the linkers and tetranuclear silver clusters, [Ag4(micro3)-Cl){S2P(OR)2}3], as the network nodes in the formation of the honeycomb-shaped layer structure is reported. PMID- 15834480 TI - Sol-gel synthesis of ordered mesoporous alumina. AB - Well-ordered mesoporous alumina materials with high surface area and a narrow pore size distribution were synthesized using a sol-gel based self assembly technique. PMID- 15834481 TI - Synthesis of 4-substituted homoallylic alcohols via a one-pot tandem Lewis-acid catalyzed crotylboration-[3,3]-sigmatropic rearrangement. AB - Crotylboration of aldehydes with E- or Z-crotylboronates in the presence of catalytic amounts of indium triflate provides the corresponding 4-substituted homoallylic alcohols. PMID- 15834482 TI - Homogeneous and heterogenized Au(III) Schiff base-complexes as selective and general catalysts for self-coupling of aryl boronic acids. AB - A series of homogeneous and heterogenized gold metal complexes show high activity and selectivity for the homocoupling of a large variety of aryl boronic acids, being of general utility for the synthesis of C2-symmetric biaryls. PMID- 15834483 TI - Towards an easy access to amphiphilic rod-coil miktoarm star copolymers. AB - Water-soluble stimuli-responsive AB2 miktoarm star copolymers were prepared by atom transfer radical polymerisation of styrene followed by chain-end modification, polymerisation of either gamma-benzyl-L-glutamate N carboxyanhydride or tert-butylacrylate and a final step of hydrolysis. PMID- 15834484 TI - An iron-catalysed chemo- and regioselective tetrahydrofuran synthesis. AB - An intermolecular ring expansion reaction of an aryl epoxide with several dienes, acrylates, enynes or styrenes under iron catalysis, generated tetrahydrofuran derivatives in a highly chemo- and regioselective fashion. The process could be used in an unprecedented way for the one step synthesis of racemic calyxolane A and calyxolane B with acceptable diastereoselectivity. PMID- 15834485 TI - Evidence for the role of organic layers in photoconductivity of organic/inorganic hybrid nanosheets as prepared by Langmuir-Blodgett methods. AB - By measuring the photoconductivity of hybrid LB films of exfoliative layered niobate and octadecylamine, it was evidenced that the film underwent a transition from an insulator to a photosemiconductor during photo-modification treatment by UV light, which was rationalized in terms of the direct contact of inorganic nanosheets achieved by the elimination of organic layers. PMID- 15834486 TI - Fluorination of double-walled carbon nanotubes. AB - Fluorine atoms are selectively attached to the sidewall of the outer shell of DWNTs without disrupting the double-layered morphology; the stoichiometry of the fluorinated DWNTs is CF(0.30). PMID- 15834487 TI - Efficient electrocatalytic oxidation of NADH at gold nanoparticles self-assembled on three-dimensional sol-gel network. AB - It is demonstrated that gold (Au) nanoparticles self-assembled on a sol-gel derived three-dimensional silicate network efficiently catalyze the oxidation of NADH in the absence of any electron transfer mediators with a decrease in overpotential of 780 mV in neutral solution. PMID- 15834488 TI - Intramolecular iodoarylation reaction of alkynes: easy access to derivatives of benzofused heterocycles. AB - The iodoarylation reaction of heteroatom-tethered omega-arylalkynes offers an efficient and straightforward entry to heterocycles. As a result, both C-C ring closing from readily available precursors, and concomitant selective iodination take place. The first related study conducted in water is presented. PMID- 15834489 TI - Scattering spherical voids in nanocrystalline TiO2- enhancement of efficiency in dye-sensitized solar cells. AB - Spherical voids as light scattering centers in nanocrystalline TiO2 films were realized with polystyrene particles of diameter 400 nm, thus enhancing the photovoltaic performance by 25% on large areas, as well as providing an indication that these films can be used with electrolytes of higher viscosity. PMID- 15834490 TI - Reactivity of a 1:1 copper-oxygen complex: isolation of a Cu(II)-o iminosemiquinonato species. AB - While a 1:1 Cu-O2 adduct is generally unreactive with organic substrates, phosphines displace O2 via an associative process and added Cu(I) leads to a novel internal ligand oxidation to yield a Cu(II)-o-iminosemiquinone complex. PMID- 15834491 TI - Catalytic metathesis of carbon dioxide with heterocumulenes mediated by titanium isopropoxide. AB - The insertion of an isopropoxide ligand of titanium isopropoxide into heterocumulenes gives a product that carries out metathesis at elevated temperatures by undergoing insertion of a second heterocumulene in a head to head fashion, followed by an extrusion reaction. PMID- 15834492 TI - A new approach to grafting a monolayer of oriented Mn12 nanomagnets on silicon. AB - The functionalisation of a Si(100) silicon wafer allows for the oriented grafting of a monolayer of Mn12 nanomagnets using a two-step procedure. PMID- 15834493 TI - RNA duplexes with biphenyl substituents as base replacements are less stable than DNA duplexes. AB - Introduction of a hydrophobic biphenyl-C-nucleotide pair into a 11-mer RNA duplex is associated with a net penalty in the free energy of duplex formation of 2.0 kcal mol(-1) or 10 degrees C in Tm, relative to DNA. These differential stabilities are of relevance with respect to the transcriptional and translational aspects of hydrophobic base-pairs. PMID- 15834494 TI - Highly efficient catalysts for the hydrogenation of nitro-substituted aromatics. AB - Nanoparticles of Co and NiPd, derived from colloidal precursors and supported on commercially available non-ordered mesoporous silica, are highly effective, cheap, recyclable and industrially viable catalysts for the hydrogenation of a range of nitro-substituted aromatics under mild conditions. PMID- 15834495 TI - New chromophores from click chemistry for two-photon absorption and tuneable photoluminescence. AB - New multipolar derivatives built from triazole moieties that combine full transparency and strong TPA in the visible region as well as fully tuneable fluorescence have been designed; such chromophores open a new route for optical limiting. PMID- 15834496 TI - Nanofiberization of inner helical mesoporous silica using chiral gelator as template under a shear flow. AB - Aligned nanofibers with inner-helical mesoporous silica were synthesized using chiral gelator as template under basic conditions and a shear flow. PMID- 15834497 TI - Carbohydrate-coated nanocapsules from amphiphilic rod-coil molecule: binding to bacterial type 1 pili. AB - Stable carbohydrate-coated nanocapsules designed as multivalent nanoscaffolds for selective interactions with receptors are able to encapsulate guest molecules within their interior and to bind efficiently to FimH adhesin of bacterial type 1 pili. PMID- 15834498 TI - Multiwalled carbon nanotubes in donor-acceptor nanohybrids-towards long-lived electron transfer products. AB - Novel multiwalled carbon nanotube/metalloporphyrin nanohybrids are devised and probed as versatile donor-acceptor hybrids. PMID- 15834499 TI - Palladium catalyzed cross-coupling reaction of 5-tributylstannyl-4 fluoropyrazole. AB - The palladium catalyzed cross-coupling reactions of aryl iodides and 5 tributylstannyl-4-fluoropyrazole prepared from fluoro(tributylstannyl)acetylene proceeded smoothly giving the corresponding 5-aryl-4-fluoropyrazole in good yields. PMID- 15834500 TI - New titanium and zirconium complexes with M-NH2 bonds formed by facile deprotonation of H3N.B(C6F5)3. AB - Facile deprotonation of H3N.B(C6F5)3 with [M(NMe2)4](M = Zr or Ti) yields the novel amidoborate complexes [Zr(NMe2)3{NH2B(C6F5)3}(HNMe2)] and [Ti(NMe2)3{NH2B(C6F5)3}]. PMID- 15834501 TI - Plausible origins of homochirality in the amino acid catalyzed neogenesis of carbohydrates. AB - The intrinsic ability of amino acids to catalyze the asymmetric formation of carbohydrates, which enzymes have mediated for millions of years, with significant amplification of enantiomeric excess suggests a plausible ancient catalytic process for the evolution of homochirality. PMID- 15834502 TI - Different firing patterns induced by veratridine and aconitine in injured dorsal root ganglion neurons. AB - Ectopic spontaneous activity originated from the injured dorsal root ganglion (DRG) neurons in rats was recorded through single dorsal root fiber. The firing patterns induced by veratridine and aconitine, inhibitors of inactivation gate of sodium channel operating on different binding sites, were compared. In the same neuron, veratridine (1.5 approximately 5.0 micromol/L) caused slow wave oscillations of interspike intervals (ISIs), while aconitine (10 approximately 200 micromol/L) caused tonic firing. Moreover, even if the background firing patterns were various and the reagent concentrations used were different, veratridine and aconitine still induced slow wave oscillations and tonic firing patterns, respectively. The results suggest that veratridine and aconitine induce different firing patterns in injured DRG neurons, which may relate to their inhibitory effects on different binding sites of the sodium channel. PMID- 15834503 TI - Fractal analysis of trabecular bone in knee osteoarthritis (OA) is a more sensitive marker of disease status than bone mineral density (BMD). AB - The purpose of this study was to determine whether fractal analysis (FSA) of macroradiographs or bone mineral density (BMD) is more sensitive in detecting disease-related cancellous bone alterations in knee osteoarthritis (OA). Differences in BMD between 11 OA (6 females) and 11 non-OA reference (7 females) tibiae were compared with differences in trabecular organization measured by computerized method of fractal signature analysis (FSA) of digitized macroradiographs (x3.5 to x5). OA knees had anatomic and radiographic evidence of medial compartment disease. FSA measured cancellous bone organization at 4 regions of interest (ROI): medial and lateral subchondral (Sc) and subarticular (Sa) sites, dual X-ray absorptiometry (DXA) measured BMD at the same ROIs. Compared to non-OA, OA tibiae had significant increased (P < 0.05) in FSA of vertical trabeculae in the medial Sa region (trabecular size range: 0.42-0.54; 0.90-1.98 mm) and significant decrease (P < 0.05) in FSA for some horizontal trabeculae in the Sc region (trabecular size range: medial side 0.12-0.18 mm; lateral side 0.12-0.24 mm). Compared to non-OA, BMD of OA tibiae was not significantly different at any ROI. BMD was not sensitive to changes in trabecular organization detected by FSA. The increase in FSA of vertical trabeculae in the medial Sa region was consistent with trabecular fenestration and thinning, which may have been detected as decreased BMD in a larger sample. For studies involving small sample sizes, quantifying changes in trabecular organization is more sensitive than BMD for detecting bone alterations in knee OA. PMID- 15834504 TI - Depressed bone mineralization following high dose TGF-beta1 application in an orthopedic implant model. AB - Several previous studies of bone repair have shown 2- to 4-fold increases in bone formation following local delivery of exogenous transforming growth factor-beta (TGF-beta). Here, we use quantitative backscatter electron microscopy to test the effect of TGF-beta1 on mineralization of regenerated bone by examining tissue samples from a previously published canine study in which we found increased bone formation. In the experiment, the proximal humeri of 10 male canines were implanted bilaterally for 28 days with porous-coated implants in the presence of a 3 mm gap between the surface of the implant and the host bone. Implants placed in the left humeri were treated with TGF-beta1 at a dose of either 120 microg (n = 5) or 335 microg (n = 5), and the implants placed in the contralateral humeri served as untreated controls. Quantitative backscatter scanning electron microscopy was used to assess the volume fraction of bone and its degree of mineralization in the 3 mm gaps. The calibrated grayscale mean and median values were depressed compared to the controls in the high dose group (p = 0.048 and p = 0.041, respectively), suggesting that high dose TGF-beta delayed or inhibited mineralization of newly formed osteoid. PMID- 15834505 TI - A new method to approach exact hemoperitoneum volume in a splenic trauma model using ultrasonography. AB - In the trauma setting, the Focused Assessment for the Sonographic examination of the Trauma patient (FAST) accurately detects hemoperitoneum. Currently, only an approximate evaluation of the volume of free intraperitoneal fluid (FIPF) can be done by imaging modalities such as ultrasound (US). The aim of this study was to correlate the thickness of FIPF measured by US in different sites of the peritoneal cavity with the total volume of an experimental post-traumatic hemoperitoneum. An intra-abdominal collection with ongoing bleeding was simulated in eight cadavers with no previous abdominal surgery. Between 200 and 2000 ml of saline solution was instilled into the left hypochondrium of eight non-embalmed cadavers. During the instillation, FIPF thickness was measured every 200 ml by US in six different declivous sites of the peritoneal cavity. The volume of FIPF instilled could be mathematically correlated with fluid thickness in all the sites through the linear equation Y=aX+b, where Y is the volume of FIPF in milliliters, a is 33 (variability coefficient), X is the FIPF thickness in millimeters and b is 470 ml (minimum volume detectable by US). The best correlation between thickness and volume was obtained in the hepatorenal pouch (Morrison pouch). Evaluation of the impact of intraperitoneal hemorrhage on the hemodynamic state of spleen trauma patients is of paramount importance for the surgeon, who has to decide whether to perform a laparotomy for hemostasis or not, specially when intra- and extra-abdominal injuries conjointly exist. After clinical validation, this new method to calculate the exact volume of FIPF could be used in current clinical practice of abdominal trauma to assist in the decision-making regarding non-operative treatment of spleen trauma. PMID- 15834506 TI - Functional analysis of PTPN11/SHP-2 mutants identified in Noonan syndrome and childhood leukemia. AB - Noonan syndrome (NS) is characterized by short stature, characteristic facial features, and heart defects. Recently, missense mutations of PTPN11, the gene encoding protein tyrosine phosphatase (PTP) SHP-2, were identified in patients with NS. Further, somatic mutations in PTPN11 were detected in childhood leukemia. Recent studies showed that the phosphatase activities of five mutations identified in NS and juvenile myelomonocytic leukemia (JMML) were increased. However, the functional properties of the other mutations remain unidentified. In this study, in order to clarify the differences between the mutations identified in NS and leukemia, we examined the phosphatase activity of 14 mutants of SHP-2. We identified nine mutations, including a novel F71I mutation, in 16 of 41 NS patients and two mutations, including a novel G503V mutation, in three of 29 patients with leukemia. Immune complex phosphatase assays of individual mutants transfected in COS7 cells showed that ten mutants identified in NS and four mutants in leukemia showed 1.4-fold to 12.7-fold increased activation compared with wild-type SHP-2. These results suggest that the pathogenesis of NS and leukemia is associated with enhanced phosphatase activity of mutant SHP-2. A comparison of the phosphatase activity in each mutant and a review of previously reported cases showed that high phosphatase activity observed in mutations at codons 61, 71, 72, and 76 was significantly associated with leukemogenesis. PMID- 15834507 TI - Molecular analysis of TBL1Y, a Y-linked homologue of TBL1X related with X-linked late-onset sensorineural deafness. AB - Recent progress in sequencing the human Y chromosome has unveiled a series of X-Y homologous genes. In the present study, we focused on Transducin beta-like 1Y (TBL1Y), which is a Y-linked homologue of TBL1X that is related with X-linked late-onset sensorineural deafness. Recently, it has been shown that TBLR1, another homologue whose gene resides on chromosome 3, and TBL1X act as a corepressor/coactivator exchanger for several nuclear receptors and transcription factors. However, the expression pattern and function of TBL1Y remain unknown. The RT-PCR analysis of the TBL1 family revealed that TBL1Y was expressed in all 13 tissues examined but not in leukocytes. Among the cell lines tested, however, it was only expressed in NT2/D1 cells and in lymphoblasts transformed with Epstein Barr (EB) virus. To compare the functions of the TBL1 family, we generated a series of expression plasmids for GAL4DBD-fused proteins of the TBL1 family. We carried out dual luciferase assays using these plasmids in combination with a plasmid having a luciferase reporter gene harboring 5xGAL4 binding sites. Unlike the other constructs, GAL4DBD-fused TBL1Y did not repress the promoter activity. Moreover, we found three novel polymorphisms in the TBL1Y gene, IVS7+9G>A, G268C, and IVS7+1G>C, which is presumed to cause splicing error. These polymorphisms are found in males within Y-haplogroup O3 (XO3e), which is defined as the Y-haplogroup O3 excluding O3e, a branch of O3. The results show that TBL1Y differs from other members of the TBL1 family in expression and function, suggesting other roles in maleness. PMID- 15834508 TI - Low RET mutation frequency and polymorphism analysis of the RET and EDNRB genes in patients with Hirschsprung disease in Taiwan. AB - Hirschsprung disease (HSCR), or congenital intestinal aganglionosis, is a relatively common disorder characterized by the absence of ganglion cells in the nerve plexuses of the lower digestive tract, resulting in intestinal obstruction in neonates. Mutations in genes of the RET receptor tyrosine kinase and endothelin receptor B (EDNRB) signaling pathways have been shown to be associated in HSCR patients. In this study, we collected genomic DNA samples from 55 HSCR patients in central Taiwan and analyzed the coding regions of the RET and EDNRB genes by PCR amplification and DNA sequencing. In the 55 patients, an A to G transition was detected in two (identical twin brothers). The mutation was at the end of RET exon 19 at codon 1062 (Y1062C), a reported critical site for the signaling pathways. Single nucleotide polymorphisms (SNP) in exons 2, 7, 11, 13, and 15 of RET and exon 4 of EDNRB in the HSCR patients or controls were detected. The differences between patients and controls in allele distribution of the five RET polymorphic sites were statistically significant. The most frequent genotype encompassing exons 2 and 13 SNPs (the polymorphic sites with the highest percentage of heterozygotes) was AA/GG in patients, which was different from the AG/GT in the normal controls. Transmission disequilibrium was observed in exons 2, 7, and 13, indicating nonrandom association of the susceptibility alleles with the disease in the patients. This study represents the first comprehensive genetic analysis of HSCR disease in Taiwan. PMID- 15834509 TI - Urethral adenocarcinoma mimicking urethral caruncle. AB - Urethral caruncles are considered as benign tumors. They need to be treated surgically; specimens should carefully be evaluated for the presence of any malignancy. We are presenting a 57-year-old female patient with a urethral lesion that looks like caruncle, but histopathologic examination of the excised lesion was reported as urethral adenocarcinoma. PMID- 15834510 TI - Is the potassium sensitivity test a valid and useful test for the diagnosis of interstitial cystitis? Against. PMID- 15834511 TI - Successful treatment of Zoon's vulvitis with high potency topical steroid. AB - Zoon's vulvitis is a rare, chronic condition of the vulva that presents with burning, pruritus, and dysuria with characteristic lesions and histopathology. Several treatment options have been reported with limited success. A 63-year-old woman with Zoon's vulvitis diagnosed on histopathology was treated with clobetasol propionate 0.05%. Complete resolution of her symptoms and lesions occurred in less than 1 week. No recurrence of her symptoms has occurred after 9 months. Zoon's vulvitis may be successfully and expeditiously treated with high potency topical steroids. PMID- 15834512 TI - Lateral vertebral assessment: a valuable technique to detect clinically significant vertebral fractures. AB - Although many vertebral fractures are clinically silent, they are associated with increased risk for subsequent osteoporotic fractures. A substantial number of these fractures are demonstrable using instant vertebral assessment with Hologic densitometers. Whether similar recognition is possible using dual-energy lateral vertebral assessment (LVA) with GE Lunar densitometers remains uncertain. Thus, we evaluated the ability of clinicians using LVA to detect prevalent vertebral fractures. Dual-energy LVA and conventional thoracic and lumbar spine radiographs were concurrently obtained in 80 postmenopausal women. Using an established visual semiquantitative system, vertebral fractures were identified individually by two non-radiologist clinicians on LVA images, and the results were compared with spinal radiograph evaluation by an expert radiologist. Using LVA, 95% of vertebral bodies from T7 through L4 were evaluable, but a majority (66%) of vertebrae from T4 to T6 were not adequately visualized. In the LVA-evaluable vertebrae, prevalent fractures were identified in 40 vertebral bodies by radiography. In this regard, the clinicians using LVA detected 17 of 18 radiographically evident vertebral fractures of grade 2 or 3, a false negative rate of 6%. They identified 50% (11/22) of grade 1 fractures. Additionally, the vast majority of evaluable non-fractured vertebrae, (764/794, 96.2%) were correctly classified as normal by LVA. Thus, clinicians utilizing LVA correctly identified the vast majority of grade 2 or 3 vertebral compression fractures and normal vertebral bodies, although detection of grade 1 fractures was less effective. In conclusion, the low-radiation, dual-energy LVA technique provides a rapid and convenient way for clinicians to identify patients with, and without, grade 2 or 3 vertebral fractures, thereby enhancing care of osteoporotic patients. PMID- 15834513 TI - Molecular cloning and phylogeny of HERV-E family that is expressed in Japanese monkey (Macaca fuscata) tissues. AB - More than 50 copies of HERV-E family have been estimated to exist in the human genome. Here, we examined the expression pattern and their relationships of the HERV-E in Japanese monkey tissues by RT-PCR and sequence analysis. The env gene of HERV-E family was expressed in monkey tissues (testis, prostate, kidney, thymus, intestine and stomach) except for cerebellum, pancreas and ovary, exhibiting that they may have transcriptional potential. Phylogenetic analysis of the HERV-E env family from Japanese monkey tissues and our previous data could be divided into two distinctive groups (I and II). They were integrated into the genomes of anthropoids and have evolved at the rate of 0.3% nucleotide differences per MYr through evolutionary divergence in primate evolution. Divergence times of the two groups were estimated as 11.6 MYr for group I and 41.6 MYr for group II. Those HERV-E sequences were extensively proliferated in the genome of humans and great apes. These data will contribute to further studies on the transcriptional potential of the HERV-E family in the Japanese monkey genome and to biomedical knowledge related to human diseases. PMID- 15834514 TI - In Memoriam. David Ogilvie White (1931-2004). PMID- 15834515 TI - Evaluation of microfluidic biosensor development using microscopic analysis of molecular beacon hybridization kinetics. AB - Molecular beacons, oligonucleotide probes that fluoresce upon hybridization to a target nucleic acid, can be used in microfluidic devices to detect and quantify nucleic acids in solution as well as inside bacterial cells. Three essential steps towards the development of such devices as integrated microfluidic biosensors using molecular beacons were investigated in the present study. First, experiments using real-time confocal microscopy indicated that diffusion of DNA molecular beacons across a 100-mum diameter microfluidic channel took less than one minute after the flow of reagents was stopped. Second, experiments to evaluate hybridization kinetics of DNA molecular beacons with target nucleic acids in solution showed that DNA molecular beacons can be used to characterize hybridization kinetics in real time in microfluidic channels and that hybridization signals approached their maximum in approximately three minutes. Finally, it was demonstrated that peptide nucleic acid molecular beacons can be used to detect bacterial cells in microfluidic devices. These results suggest that the use of microfluidic devices to detect nucleic acids in solution and in bacterial cells is promising and that further development of an integrated microfluidic biosensor for bacterial detection based on this concept is warranted. PMID- 15834516 TI - Rapid mixing using two-phase hydraulic focusing in microchannels. AB - Rapid mixing is important in biomedical analysis. In this study, rapid mixing is obtained through two-phase hydraulic focusing in microchannels. Two mixing streams are focused by two sheath streams. Assuming a laminar flow in the channel, the spreading behavior of the two immiscible fluids is modeled and solved analytically. The results show that both viscosity ratio and flow rate ratio between the sheath flow and the sample flow can affect the focusing ratio. Thus, the mixing path of the sample flows can be adjusted by either viscosity ratio or flow rate ratio. Furthermore, an analytical model was proposed and solved for convective/diffusive mixing between the sample streams. According to this model, the focusing ratio is a key parameter for rapid mixing. A fully polymeric micro mixer was fabricated and tested for verification of the presented analytical models. The micromixer was fabricated by laser micromachining and adhesive bonding. The characterization results show the promising potential of mixing in microscale using two-phase hydraulic focusing. PMID- 15834517 TI - A theoretical model of a molecular-motor-powered pump. AB - The motion of a cylindrical bead in a fluid contained within a two-dimensional channel is investigated using the boundary element method as a model of a biomolecular-motor-powered microfluidics pump. The novelty of the pump lies in the use of motor proteins (kinesin) to power the bead motion and the few moving parts comprising the pump. The performance and feasibility of this pump design is investigated using two model geometries: a straight channel, and a curved channel with two concentric circular walls. In the straight channel geometry, it is shown that increasing the bead radius relative to the channel width, increases the flow rate at the expense of increasing the force the kinesins must generate in order to move the bead. Pump efficiency is generally higher for larger bead radii, and larger beads can support higher imposed loads. In the circular channel geometry, it is shown that bead rotation modifies the force required to move the bead and that shifting the bead inward slightly reduces the required force. Bead rotation has a minimal effect on flow rate. Recirculation regions, which can develop between the bead and the channel walls, influence the stresses and force on the bead. These results suggest this pump design is feasible, and the kinesin molecules provide sufficient force to deliver pico- to atto- l/s flows. PMID- 15834518 TI - Micro/Nanofluidic device for single-cell-based assay. AB - In this paper we have proposed and demonstrated a microfluidic device for a fast and parallel single-cell based assay. The proposed device is designed to passively capture single-cells or beads on multiple cell-positioning sites by a pre-defined fluidic stream and inject specific reagents or drugs onto each isolated single-cell. The device consists of surface-modified silicon channels capped with a grooved polydimethylsiloxane (PDMS) cover layer. A cell capture experiment has been performed using polystyrene beads as well as CHO DG44 living cells, and successful positioning of a single-cell on the cell-positioning sites was achieved. Also, we have demonstrated independent drug injection into a specific target cell. PMID- 15834519 TI - Micropatterned structures for studying the mechanics of biological polymers. AB - Studying the mechanics of nanometer-scale biomolecules presents many challenges; these include maintaining light microscopy image quality and avoiding interference with the laser used for mechanical manipulation, that is, optical tweezers. Studying the pushing forces of a polymerizing filament requires barriers that meet these requirements and that can impede and restrain nanoscale structures subject to rapid thermal movements. We present a flexible technique that meets these criteria, allowing complex barrier geometries with undercut sidewall profiles to be produced on #1 cover glass for the purpose of obstructing and constraining polymerizing filaments, particularly microtubules. Using a two layer lithographic process we are able to separate the construction of the primary features from the construction of a depth and shape-controlled undercut. The process can also be extended to create a large uniform gap between an SU-8 photoresist layer and the glass substrate. This technique can be easily scaled to produce large quantities of shelf-stable, reusable microstructures that are generally applicable to microscale studies of the interaction of cellular structures with defined microscale features. PMID- 15834520 TI - In vitro and in vivo characterization of MEMS microneedles. AB - Transdermal drug delivery TDD systems have many advantages but are conventionally limited by the low permeability of skin. The idea of using microneedles to painlessly penetrate the topmost impermeable stratum corneum has previously been put forward. In this paper, the fabrication of solid and hollow silicon microneedles with straight side-walls and with the following dimensions: 20-100 microm in diameter and 100-150 microm in length is described. In vitro tests demonstrate that with prior solid microneedle application, transdermal drug transport is significantly increased by 10-20 times, with the degree of enhancement being related to needle diameter. In vivo tests in diabetic animals, however, were unable to demonstrate any delivery of insulin through the hollow microneedles. It is proposed that two factors, microneedle length and tip sharpness, have to be improved for systemic drug delivery to be seen in vivo. PMID- 15834521 TI - Evaluation of time-course changes of gingival crevicular fluid glucose levels in diabetics. AB - A minute volume such as a few hundred nano-litres of gingival crevicular fluid (GCF) will evaporate within one minute at room temperature after collection. In order to investigate the feasibility of a method for noninvasive blood glucose measurement using this fluid, a fabricated GCF-collecting device, and the time course changes of blood and GCF glucose levels in diabetics were evaluated. As a result of improvement in the GCF-collecting device such that the color of a filter paper changed from white to black upon collection of the sample, the completion of the collecting procedure could be visually confirmed. Using this device, we succeeded in collecting 200 nl of GCF within 30 s, despite the fact that GCF is usually secreted at a rate between 2 and 3 microl per tooth per hour. This method could be applicable for collecting small volume of other biological fluids. The time-course changes of blood and GCF glucose levels in ten type 2 diabetic subjects were examined, the correlation coefficient, R, between both glucose levels was 0.878. The result proved to be similar to that for normal healthy subjects in a previous study. A significant time difference between the two glucose levels was not observed. In diabetes period of 4.8 +/- 1.7 years, the influence of the disease on the GCF glucose level was minimal. Furthermore, from the ROC curve analysis, both sensitivity and specificity showed favorable results. It was suggested that the measurement of GCF glucose levels was a promising screening method for diabetics. PMID- 15834522 TI - Microdialysis microneedles for continuous medical monitoring. AB - Enzyme based biosensors suffer from loss of activity and sensitivity through a variety of processes. One major reason for the loss is through large molecular weight proteins settling onto the sensor and affecting sensor signal stability and disrupting enzyme function. One way to minimize loss of sensor activity is to filter out large molecular weight compounds before sensing small biochemicals such as glucose. A novel microdialysis microneedle is introduced that is capable of excluding large MW compounds based on size. Preliminary experimental evidence of membrane permeability is shown, as well as diffusion and permeability modeling. Microdialysis microneedles present an attractive first step towards decreasing size, patient discomfort and energy consumption of portable medical monitors over existing technologies. PMID- 15834523 TI - Release of biologically functional interferon-alpha from a nanochannel delivery system. AB - Metastatic melanoma lesions often are unresectable due to their size and/or location near critical structures. These lesions represent a significant challenge for the oncologist, because radiation therapy and chemotherapy are infrequently successful in halting tumor growth. Of primary concern is the fact that these lesions are usually painful and present a cosmetic dilemma. We hypothesized that the development of a silicon-based nano-device capable of delivering antitumor compounds (e.g. immune modulators), locally, at a constant rate, to the tumor microenvironment could avoid the toxicity of systemic administration and the inconvenience of frequent clinic visits for local injections. Because of its diminutive size, such a device could be implanted using a minimally invasive procedure in close proximity to unresectable melanoma lesions. The current report uses interferon alpha-2b (IFN-alpha) as a model antitumor agent, since it is commonly used in the treatment of malignant melanoma and metastatic renal cell carcinoma. In this system, IFN-alpha is delivered directly to the tumor microenvironment by a novel nanochannel delivery system (nDS) that is capable of zero order release of small molecules. We have demonstrated that the IFN-alpha released from the nDS is functionally active on both host immune cells and a human melanoma cell line in vitro. This drug delivery platform could be used to develop alternative strategies for the treatment of unresectable tumors. PMID- 15834524 TI - [Opening speech of the President at the 111. Meeting of the DGIM (German Society of Internal Medicine). Changing doctor-patient relations]. PMID- 15834525 TI - [Structure of an interdisciplinary pituitary outpatient care unit at the University Hospital of Leipzig and results for treatment of prolactin and growth hormone secreting pituitary tumors]. AB - BACKGROUND: Treatment of patients with pituitary adenomas is complex and involves several medical specialties. At the Medical Center of the University of Leipzig, Germany, an interdisciplinary pituitary outpatient care unit has been established for 6 years. METHODS: The interdisciplinary collaboration and the outcome of patients with growth hormone-(GH-) and prolactin-secreting pituitary adenomas are described. Moreover, therapeutic strategies for patients with hormonally active pituitary adenomas are presented and discussed. RESULTS: In patients suffering from GH-producing adenomas, a remission could be achieved in 80% (microadenomas) and 40% (macroadenomas) of the cases, respectively. This is comparable to major published studies. Furthermore, prolactinomas decreased in size during treatment in at least 75% of all cases depending on the initial size of the lesion which is also comparable to data from other groups. CONCLUSION: Taken together, an interdisciplinary approach improves outcome and quality of care of patients with hormonally active pituitary adenomas. PMID- 15834526 TI - [Long-term outcome of intrapleural fibrinolytic therapy via small-bore catheter drainage in the management of complicated parapneumonic effusion and empyema-a case series]. AB - BACKGROUND AND PURPOSE: Complicated parapneumonic effusions (PPE) and pleural empyemas (PE) are associated with increased morbidity and mortality. Intrapleural fibrinolytic therapy (IFT) has been established, besides video-assisted thoracoscopy (VATS), in the management of PPE and PE. Combination of IFT and small-bore catheter drainage has previously not been investigated. PATIENTS AND METHODS: 15 patients consecutively referred for PPE and PE were managed with simultaneous IFT via small-bore catheter drainage (9, 12, 14 F). In addition to evaluation of primary outcome, lung function testing and radiologic studies were obtained after 3 months to predict potential pulmonary restrictive impairment and residual pleural thickening. RESULTS: Primary outcome of all 15 patients was positive. Only one patient showed a significant residual pleural thickening. None of the patients showed a restrictive lung function pattern. CONCLUSION: Proven applicability of IFT via small-bore catheter drainage seems to be effective in the face of long-term outcome. Further prospective and multicentric studies should be initiated to investigate the practicability of IFT via small-bore catheter drainage. PMID- 15834527 TI - [Niacin--an additive therapeutic approach for optimizing lipid profile]. AB - BACKGROUND: Large interventional studies have shown that the reduction of total cholesterol and low-density lipoprotein cholesterol (LDL-C) is one of the cornerstones in the prevention of coronary artery disease. However, in up to 40% of patients the recommended target of LDL-C is not reached with a monotherapy. Furthermore, risk stratification only by LDL-C disregards a substantial number of patients with dyslipidemia with increased triglycerides and decreased high density lipoprotein cholesterol (HDL-C). EFFECT OF NIACIN ON LIPID METABOLISM: In consequence, niacin has gained attention as a component of a combined therapeutic approach in patients with dyslipidemia. Niacin substantially increases HDL-C and decreases triglycerides, LDL-C and lipoprotein (a). By this mechanism of action niacin exhibited, in combination with statins or bile acid-binding resins, favorable effects on the incidence of cardiovascular events in selected patients. Side effects like flush and hepatotoxicity seem to be in part dependent on the niacin formulations used. However, niacin has been shown to be a well-tolerated and safe therapy in controlled studies. CONCLUSION: On the basis of current data niacin should be considered a valuable therapy component in patients with dyslipidemia, in which a monotherapy fails to optimize an increased risk of coronary artery disease. PMID- 15834528 TI - [Shared decision-making as a new quality indicator in nephrology: a nationwide survey in Germany]. AB - BACKGROUND AND PURPOSE: Shared decision-making (SDM) as a model in physician patient interaction is gaining relevance in the German health system. By applying this model, mid- and long-term improvements are expected especially in the outcomes of chronic diseases. Up to now, there has hardly been any empirical data available in German health services research regarding the state and development of SDM. This study establishes a baseline and provides actual data on this subject based on a German-wide survey of end-stage renal disease (ESRD) patients. METHODS: Standardized German-wide survey of 6,614 patients with ESRD. The questionnaire included an instrument to measure the patients' perceived involvement in care (PICS) which had been translated and validated before. RESULTS: 82% of the questioned patients feel their physicians facilitated involvement in decision making. 81% of the patients actively inform themselves concerning their disease and treatment options. 69% state that SDM has taken place. Age, years on dialysis and gender correlate with perceived involvement. CONCLUSION: This paper provides a valid baseline for the prospective research of SDM in ESRD. The results indicate that dialysis patients are willing to participate in the process of medical decision-making. Characteristics and preferences of the patients should be taken into account not only in everyday clinical interactions. They could be monitored systematically within the framework of quality management and used as potential for quality improvement. PMID- 15834529 TI - [Treatment of resistant hypertension]. PMID- 15834530 TI - [Atypical headache and facial pain as a result of hypertrophic pachymeningitis in C-ANCA-positive Wegener's granulomatosis]. AB - BACKGROUND: Wegener's granulomatosis (WG) is a systemic vasculitis involving the nervous system in 20-54% of cases; lesions of peripheral nerves are commonest, while manifestation in the central nervous system (CNS) is rarer. Focal hypertrophic pachymeningitis is a very rare complication of WG. This inflammatory thickening and fibrosis of the dura mater is always associated with headaches, whereas cranial nerve lesions, cerebellar symptoms or epileptic seizures occur more rarely. CASE REPORT: A 67-year-old patient, in whom WG had been diagnosed 2 years earlier and who had been treated with immunosuppressants since then, complained of continuous severe, mainly left-sided headache and facial pain for weeks. Cranial MRI showed thickening of the left tentorium cerebelli with obvious contrast enhancement and led to the diagnosis of hypertrophic pachymeningitis. The inflammatory parameters and the C-ANCA (antineutrophil cytoplasmic antibodies) in the serum were raised and CANCA were detectable in the cerebrospinal fluid. The headaches subsided with several days of intravenous high dose corticosteroids and a simultaneous increase in the immunosuppressive basic medication. On a follow-up MRI after 3 months, the magnetic resonance changes were less apparent, i. e., the hypertrophic pachymeningitis was resolving; C-ANCA were now no longer detectable in the cerebrospinal fluid. CONCLUSION: With newly occurring, unusually severe and persistent headaches in the presence of WG, the very rare complication of hypertrophic pachymeningitis should be considered. PMID- 15834532 TI - [Pitfalls in DRG coding: HIV and AIDS]. PMID- 15834531 TI - [Extreme sinus bradycardia (30/min) with acute right heart failure under tizanidine (Sirdalud). Possible pharmacological interaction with rofecoxib (Vioxx)]. AB - CASE RECORD: The case of a 59-year-old healthy woman is described, who developed an extreme sinus bradycardia (30/min) with chest pain and acute right heart failure associated with gastrointestinal symptoms and elevation of the liver enzymes while simultaneously taking tizanidine (Sirdalud), diclofenac (Voltaren), and rofecoxib (Vioxx). The symptomatology resolved promptly after stopping the medication. DISCUSSION: The usual causes of sinusbradycardia like hypothyroidism, hypothermia, intracranial pressure elevation, typhoid fever, sick sinus syndrome, hyperreactive carotid sinus reflex, organic heart disease, electrolyte disorders, and pharmacotherapy with beta-blockers, digitalis, and antiarrhythmics have been excluded in this case. Bradycardia can occur as a side effect of tizanidine. As this substance is metabolized by cytochrome P450 1A2 and rofecoxib inhibits this enzyme, an interaction between these drugs is probable. Liver function disorders and gastrointestinal symptoms, in the present case mainly due to the acute right heart failure, have also been described as side effects under tizanidine, diclofenac as well as rofecoxib. Supposedly, the combination of tizanidine/rofecoxib used to be prescribed frequently for lumbar pain as selective cyclooxygenase-(COX-)2 inhibitors are visibly replacing the nonsteroidal antirheumatics due to their better side effect profile. An augmented risk of cardiovascular events under rofecoxib led to its withdrawal from the market at the end of September 2004. RESULTS: When prescribing Sirdalud, the possible pharmacological side effects and interactions should be taken into careful account. PMID- 15834533 TI - [Indications for cardiovascular magnetic resonance imaging]. AB - Cardiovascular magnetic resonance (CMR) has moved from a tool mainly used for congenital heart disease, large vessels, pericardium and tumors to a flexible, highly accurate and rapid method with a broad range of indications. New applications are especially found in patients with ischemic heart disease, ranging from dobutamine stress and adenosine perfusion to scar imaging, viability testing and-increasingly-visualization of the coronary arteries themselves. PMID- 15834534 TI - [Late metastasis of granulosa cell tumor in a 74-year-old patient]. PMID- 15834537 TI - Alcohol increases cognitive biases for smoking cues in smokers. AB - RATIONALE: Alcohol increases the motivation to smoke, possibly because it increases the incentive motivational properties of smoking cues. OBJECTIVES: We examined whether alcohol would increase attentional, approach, and evaluative biases for smoking-related cues in a sample of daily cigarette smokers. METHODS: The study used a visual probe task with eye movement recording to investigate biases in visual orienting to smoking-related cues. A stimulus-response compatibility task was used to assess approach tendencies for smoking-related cues, and an explicit rating task was used to assess the perceived valence of smoking-related cues. Participants completed the tasks in two sessions, once after consumption of 0.4 g/kg alcohol and once after consumption of a non alcoholic drink. RESULTS: Alcohol increased the maintenance of attention on smoking cues (evident from gaze duration and a reaction time index of attentional bias), the perceived pleasantness of smoking cues, and cigarette craving, relative to the non-alcoholic drink. However, alcohol had no effect on the initial shifting of gaze to smoking cues or on the tendency to approach smoking cues. CONCLUSIONS: These results suggest that, in smokers, ingestion of a moderate dose of alcohol increases the propensity for smoking-related cues to hold attention and makes those cues seem more attractive, which is consistent with alcohol increasing the 'incentive salience' of smoking cues. PMID- 15834536 TI - Neurocircuitry in alcoholism: a substrate of disruption and repair. AB - The chronic, excessive consumption of alcohol results in significant modification of selective neural systems of the brain structure, physiology, and function. Quantitative MR structural imaging, diffusion tensor imaging (DTI), and functional MRI (fMRI), together with neuropsychological challenges, have enabled rigorous in vivo characterization of the results of alcoholism on the brain in the human condition. Neuroimaging has also enabled longitudinal study for the examination of alcoholism's dynamic course through periods of drinking and sobriety. Controlled studies have revealed compelling evidence for alcohol related brain structural and functional modification--some longstanding, some transient, and some compensatory. Patterns of circuitry disruption identified through structural and functional MRI studies suggest a central role for degradation of frontocerebellar neuronal nodes and connecting circuitry affecting widespread brain regions and contributing to alcoholism's salient, enduring, and debilitating cognitive and motor deficits--executive dysfunction, visuospatial impairment, and ataxia. PMID- 15834538 TI - 5-HT2A/2C receptor signaling via phospholipase A2 and arachidonic acid is attenuated in mice lacking the serotonin reuptake transporter. AB - SUBJECTS: The serotonin reuptake transporter (SERT) helps to regulate brain serotonergic transmission and is the target of some antidepressants. To further understand SERT function, we measured a marker of regional brain phospholipase A2 (PLA2) activation in SERT knockout mice (SERT-/-) and their littermate controls (SERT+/+). METHODS: Following administration of 1.5 mg/kg s.c. (+/-)-2,5 dimethoxy-4-iodophenyl-2-aminopropane (DOI), a 5-HT(2A/2C) receptor agonist, to unanesthetized mice injected intravenously with radiolabeled arachidonic acid (AA), PLA2 activation, represented as the regional incorporation coefficient k* of AA, was determined with quantitative autoradiography in each of 71 brain regions. RESULTS: In SERT+/+ mice, DOI significantly increased k* in 27 regions known to have 5-HT(2A/2C) receptors, including the frontal, motor, somatosensory, pyriform and cingulate cortex, white matter, nucleus accumbens, caudate putamen, septum, CA1 of hippocampus, thalamus, and hypothalamus. In contrast, DOI did not increase k* significantly in any brain region of SERT-/- mice. Head twitches following DOI, which also were measured, were robust in SERT+/+ mice but were markedly attenuated in SERT-/- mice. CONCLUSIONS: These results show that a lifelong elevation of the synaptic 5-HT concentration in SERT-/- mice leads to downregulation of 5-HT(2A/2C) receptor-mediated PLA2 signaling via AA and of head twitches, in response to DOI. PMID- 15834539 TI - Attenuated adrenocorticotropic responses to psychological stress are associated with early smoking relapse. AB - RATIONALE: Research has demonstrated that psychosocial stressors increase smoking and risk for smoking relapse. Alterations in biological systems involved in the stress response caused by chronic smoking may contribute to early relapse. OBJECTIVES: We examined the extent to which pituitary-adrenocortical and cardiovascular responses to stress following the first 24 h of a quit attempt predict early relapse. METHODS: Seventy-two smokers interested in cessation attended a laboratory stress session 24 h after the beginning of their cessation attempt. Adrenocorticotropin (ACTH), plasma and salivary cortisol concentrations, systolic and diastolic blood pressure (BP), and heart rate (HR) responses to acute psychological stressors (public speaking and cognitive challenges) were used to predict relapse over a 4-week follow-up period. RESULTS: Those who relapsed within 4 weeks showed attenuated hormonal and cardiovascular responses to stress and exaggerated withdrawal symptoms. Cox proportional hazards survival analysis showed that attenuated ACTH, plasma cortisol, systolic and diastolic BP, positive affect, and exaggerated withdrawal symptoms and smoking urges during acute stress predicted early relapse. Stepwise model showed that ACTH, diastolic BP, and exaggerated withdrawal symptoms remain as significant predictors. When baseline smoking and psychological measures were included in the model, changes in ACTH, diastolic BP, and both factors of smoking urges remained significant predictors of relapse. CONCLUSIONS: These results demonstrate that altered stress response predicts increased vulnerability for smoking relapse. PMID- 15834540 TI - [Optimization of cholesterol reduction principles and clinical results of dual inhibition]. AB - Ezetimibe is a recently developed compound, which inhibits intestinal cholesterol absorption. Because there are hints for an increase of cholesterol absorption during statin therapy, the combination of Ezetimibe with a statin seems to be appropriate. This dual approach -- inhibition of intestinal cholesterol absorption and hepatic cholesterol synthesis -- offers a very potent reduction of cholesterol. The combination of statins with Ezetimibe leads to a further reduction of LDL-cholesterol up to 12-21%. The dual inhibition causes a more effective reduction of LDL-cholesterol than a statin monotherapy. LDL treatment goals can be reached more easily, and possible side effects of otherwise necessary high doses of statins can be avoided. Clinical endpoint studies with Ezetimibe are underway. PMID- 15834542 TI - [Peace-study. Coronary heart disease: additive therapy with ACE-inhibitors?]. PMID- 15834543 TI - The sea-finding behavior of hatchling olive ridley sea turtles, Lepidochelys olivacea, at the beach of San Miguel (Costa Rica). AB - Newly hatched olive ridley sea turtles (Lepidochelys olivacea) were tested for their directional preferences in a sand-filled circular arena in total darkness. Hatchlings that had crawled about 5 m on the beach, toward the sea preferred the southwesterly direction that would have brought them to the water line, whereas hatchlings that had been denied this experience headed eastward, a direction of unclear origin. These data suggest that a short crawl across the natural beach can set the direction in which the young turtles subsequently move. The crawling experience was sufficient to acquire the compass course that they later follow, probably with the help of a magnetic compass, not only in the water, but already while still on land. PMID- 15834541 TI - [Blood pressure and the brain]. AB - During acute cerebral infarction, autoregulation is abolished. Brain perfusion therefore directly depends on perfusion pressure and cardiac output. For this reason, in the early state of stroke, elevated blood pressure improves cerebral blood flow and only values of 210 mmHg systolic or above should be lowered. With the development of a vasogenic brain edema or a dysfunctional blood-brain barrier (usually on day 2 to 4 after infarction), blood pressure must be normalized in order to avoid hemorrhage and to minimize edema. In the presence of space occupying edema or intracranial hemorrhage, only those antihypertensive substances may be used which do not cause a dilatation of brain vessels. Direct vasodilators and calcium antagonists are not suitable in this situation. Furthermore, antihypertensive medication which causes bradycardia (e.g. beta blockers) should be avoided, because in acute stroke, brain perfusion also depends on the cardiac output. For primary and secondary stroke prevention normalization of blood pressure is essential. Efficacy is basically independent of the kind of antihypertensive medication used. Effective normalization of blood pressure probably helps to prevent vascular dementias of all kinds. Convincing studies however are still lacking for most sorts of antihypertensive medication. PMID- 15834544 TI - Strain-specific and recessive QTLs involved in the control of partial resistance to Fusarium oxysporum f. sp. melonis race 1.2 in a recombinant inbred line population of melon. AB - Fusarium oxysporum f. sp. melonis (FOM) causes serious economic losses in melon (Cucumis melo L.). Two dominant resistance genes have been identified, Fom-1 and Fom-2, which provide resistance to races 0 and 2 and races 0 and 1, respectively, however FOM race 1.2 overcomes these resistance genes. A partial resistance to FOM race 1.2 that has been found in some Far East accessions is under polygenic control. A genetic map of melon was constructed to tag FOM race 1.2 resistance with DNA markers on a recombinant inbred line population derived from a cross between resistant (Isabelle) and susceptible (cv. Vedrantais) lines. Artificial root inoculations on plantlets of this population using two strains, one that causes wilting (FOM 1.2w) and one that causes yellowing (FOM 1.2y), resulted in phenotypic and genotypic data that enabled the identification of nine quantitative trait loci (QTLs). These QTLs were detected on five linkage groups by composite interval mapping and explained between 41.9% and 66.4% of the total variation. Four digenic epistatic interactions involving seven loci were detected and increased the total phenotypic variation that was explained. Co-localizations between QTLs and resistance gene homologs or resistance genes, such as Fom-2 and Vat, were observed. A strain-specific QTL was detected, and some QTLs appeared to be recessive. PMID- 15834545 TI - Serum concentrations of resistin-like molecules beta and gamma are elevated in high-fat-fed and obese db/db mice, with increased production in the intestinal tract and bone marrow. AB - AIMS/HYPOTHESIS: Resistin and the resistin-like molecules (RELMs) comprise a novel class of cysteine-rich proteins. Among the RELMs, RELMbeta and RELMgamma are produced in non-adipocyte tissues, but the regulation of their expression and their physiological roles are largely unknown. We investigated in mice the tissue distribution and dimer formation of RELMbeta and RELMgamma and then examined whether their serum concentrations and tissue expression levels are related to insulin resistance. METHODS: Specific antibodies against RELMbeta and RELMgamma were generated. Dimer formation was examined using COS cells and the colon. RELMbeta and RELMgamma tissue localisation and expression levels were analysed by an RNase protection assay, immunoblotting and immunohistochemical study. Serum concentrations in high-fat-fed and db/db mice were also measured using the specific antibodies. RESULTS: The intestinal tract produces RELMbeta and RELMgamma, and colonic epithelial cells in particular express both RELMbeta and RELMgamma. In addition, RELMbeta and RELMgamma were shown to form a homodimer and a heterodimer with each other, in an overexpression system using cultured cells, and in mouse colon and serum. Serum RELMbeta and RELMgamma levels in high-fat-fed mice were markedly higher than those in mice fed normal chow. Serum RELMbeta and RELMgamma concentrations were also clearly higher in db/db mice than in lean littermates. Tissue expression levels revealed that elevated serum concentrations of RELMbeta and RELMgamma are attributable to increased production in the colon and bone marrow. CONCLUSIONS/INTERPRETATION: RELMbeta and RELMgamma form homo/heterodimers, which are secreted into the circulation. Serum concentrations of RELMbeta and RELMgamma may be a novel intestinal-tract-mediating regulator of insulin sensitivity, possibly involved in insulin resistance induced by obesity and a high-fat diet. PMID- 15834546 TI - Effectiveness of frequency-modulated electromagnetic neural stimulation in the treatment of painful diabetic neuropathy. AB - AIMS/HYPOTHESIS: The largely unsatisfactory results reported for the pharmacological treatment of diabetic neuropathy has spurred the search for alternative therapies. The aim of this study was to evaluate the efficacy of frequency-modulated electromagnetic neural stimulation (FREMS) as a novel treatment for painful diabetic neuropathy. METHODS: Patients (n=31) with painful neuropathy associated with decreased nerve conduction velocity (<40 m/s) and increased vibration perception threshold (>25 V) were enrolled in a randomised, double-blind, crossover study designed to compare the effects of FREMS with those of placebo. Each patient received two series of ten treatments of either FREMS or placebo in random sequence, with each series lasting no more than 3 weeks. The primary efficacy end point was the change in pain measured by a visual analogue scale (VAS). RESULTS: FREMS induced a significant reduction in daytime and night time VAS pain score (all p<0.02). Furthermore, FREMS induced a significant increase in sensory tactile perception, as assessed by monofilament; a decrease in foot vibration perception threshold, as measured by a biothesiometer; and an increase in motor nerve conduction velocity (all p<0.01). No significant changes were observed after placebo. Comparison of measurements at the 4-month follow-up with those at baseline revealed that a significant benefit persisted for all measures that showed an improvement at the end of treatment, with an additional improvement in quality of life evaluated by the Short Form-36 questionnaire (all p<0.05). No significant side effects were recorded during the study. CONCLUSIONS/INTERPRETATION: FREMS is a safe and effective therapy for neuropathic pain in patients with diabetes and is able to modify some parameters of peripheral nerve function. PMID- 15834547 TI - Dynamic change in adiposity from fetal to postnatal life is involved in the metabolic syndrome associated with reduced fetal growth. AB - AIMS/HYPOTHESIS: The aims of this study were to establish the role of insulin resistance in the metabolic syndrome associated with restricted fetal growth and to characterise the fetal and postnatal determinants responsible for the long term metabolic outcome. METHODS: The study population consisted of adults selected on birth data from a maternity registry and born either small for gestational age (SGA) (n=734, birthweight20 microM) in DU145 and T24 cells. Nitrite did not inhibit the growth of SKRC cells at any of the concentrations used. Doxorubicin (DXR) inhibited cell growth in the three cell lines, whereas growth inhibition recovered in the presence of <10 microM nitrite. The recovery of DXR-induced growth inhibition was closely associated with an increase in Bcl-2 in the presence of <10 microM nitrite. Vascular endothelial growth factor (VEGF) secretion was also increased in the presence of <10 and <20 microM nitrite, respectively, in DU145 and SKRC or T24 cells. The expression of HO-1 was associated with sensitivity to NO-induced growth inhibition at constitutive levels, and was induced by SNP treatment. HO-1 inhibition by HO-1 antisense S oligodeoxynucleotide treatment increased NO-induced growth inhibition, and decreased Bcl-2 expression or VEGF secretion in the three cell lines. These findings suggest that the NO/HO-1 system has protumoral effects. PMID- 15834588 TI - De novo t(7;10)(q33;q23) translocation and closely juxtaposed microdeletion in a patient with macrocephaly and developmental delay. AB - We have applied FISH with fully integrated BACs and BAC subfragments assessed in the human genome sequence to a de novo t(7;10)(q33;q23) translocation in a patient with developmental delay and macrocephaly. The translocation breakpoints disrupt the SEC8L1 gene on chromosome 7 and the PTEN gene on chromosome 10. RT PCR demonstrated chimeric transcripts containing the first 11 exons of SEC8L1 fused to exon 3 of PTEN. In addition to the balanced translocation, we found a 7 Mb deletion in the translocated part of chromosome 7 at 4-Mb distance of the translocation breakpoint. This microdeletion, which disrupts the PTN and TPK1 genes and deletes 29 bonafide genes and the T-cell receptor beta locus, arose in the paternal germline. The patient's phenotype may be caused by a dominant negative effect of the SEC8L1-PTEN fusion protein and/or haploinsufficiency of the disrupted or deleted genes. Our study demonstrates that de novo translocations can be associated with microdeletions outside the breakpoint region(s), rendering the study and risk estimation of such breakpoints more complicated than previously assumed. PMID- 15834589 TI - Variation in ITGB3 has sex-specific associations with plasma lipoprotein(a) and whole blood serotonin levels in a population-based sample. AB - A recent genome-scan identified the Leu33Pro polymorphism in the beta3 integrin (ITGB3) gene as a quantitative trait locus for whole blood serotonin level in a large Hutterite pedigree. Because both the Leu33Pro polymorphism and the serotonin system have been implicated in cardiovascular disease (CVD) risk and treatment response, we studied additional variation in ITGB3 and its relationship to intermediate phenotypes associated with CVD in the same population. We examined associations between 15 single nucleotide polymorphisms (SNPs) across ITGB3 and five CVD-related traits in the Hutterites: plasma levels of high density lipoprotein-cholesterol (HDL-c), triglycerides (TG), low density lipoprotein-cholesterol (LDL-c), and lipoprotein(a) [Lp(a)] and blood pressure or hypertension. Seven of these SNPs in ITGB3 were associated with whole blood serotonin. Among the intermediate CVD-related phenotypes, only Lp(a) was associated with multiple ITGB3 SNPs, five of which were also associated with serotonin. A sex-stratified analysis revealed that the association between ITGB3 and Lp(a) is present only in females, whereas the association between ITGB3 and serotonin is concentrated in males. Our results suggest that variation in ITGB3 in addition to Leu33Pro could contribute to susceptibility to CVD and serotonin in a sex-specific manner. PMID- 15834590 TI - Impact of emesis on clinical and economic outcomes of cancer therapy with highly emetogenic chemotherapy regimens: a retrospective analysis of three clinical trials. AB - OBJECTIVE: It is a current hypothesis that chemotherapy-induced nausea and vomiting (CINV) may ultimately impede the clinical success of cancer treatments by hindering patients' adherence to the optimal treatment schedule. The aim of this study is to examine clinical trial data retrospectively for possible evidence of such a detrimental impact of CINV. PATIENTS AND METHODS: Data from three recent European Organization for Research and Treatment of Cancer (EORTC) trials of highly emetogenic cisplatin-based chemotherapy in diverse patient populations were analyzed retrospectively for incidence and possible impact of CINV. Data on the incidence of emesis are presented as simple descriptive analyses, while the hypothetical impact of CINV on clinical outcomes and on the patients' length of hospital stays is analyzed by means of multivariate regression analysis techniques to control for confounding variables. MAIN RESULTS: Between 42 and 59% of the patients in the trials experienced at least one episode of nausea of NCIC grade 2 or worse, while the incidence of vomiting of similar grade was between 31 and 58%. Only in one of the trials could the determinants of the adherence to protocol therapy be assessed, statistically significant variables were the severity of emesis (p < 0.0001) and other toxicities combined (p < 0.019). In turn, a Cox regression showed adherence to protocol therapy and other toxicities as the only statistically significant determinants of overall survival. CONCLUSIONS: This study has shown a discernible detrimental impact of CINV on patients' adherence to protocol therapy and, indirectly, on survival in one of the three trials examined. Further studies are required to substantiate this finding. PMID- 15834591 TI - Psychometric properties of the functional rating index in patients with low back pain. AB - MAIN PROBLEM: The purpose of this study was to validate the psychometric properties of the functional rating index (FRI), establish the instrument's minimum clinically important difference (MCID), and compare its psychometric properties with the Oswestry questionnaire. METHODS: This was a cohort study of patients with low back pain (LBP) undergoing physical therapy. One thirty one patients with a primary complaint of LBP participating in a clinical trial were assessed at baseline and at a 1- and 4-week follow-up. Test-re-test reliability was examined using the intraclass correlation coefficient, and validity was examined by determining the association between the FRI and Oswestry, a concurrent measure of disability. Responsiveness was examined by calculating the standard error of the measure, minimum detectable change, area under a receiver operating characteristic curve, and minimum clinically important difference. Changes in clinical status at each follow-up period were compared to the average of the patient and therapist's perceived improvement using the 15-point global rating of change scale. RESULTS: Test-retest reliability of the FRI was moderate, with an intraclass correlation coefficient equal to 0.63 (0.35, 0.80). Validity of the FRI was supported by a moderate correlation between the FRI and Oswestry (r=0.67, P<0.001). Area under the curve for the FRI was 0.93 (0.89, 0.98), and the minimum clinically important difference was approximately nine points. CONCLUSIONS: The FRI is less reliable than the Oswestry but appears to have comparable validity and responsiveness. Before the FRI can be recommended for widespread use in patients with neck and low back pain, it should be further tested in patients with neck pain. PMID- 15834592 TI - Does raloxifene treatment influence back pain and disability among postmenopausal women with osteoporosis? AB - Clinical studies have suggested that postmenopausal women on estrogen replacement treatment are more likely to experience back pain and related disability compared to women who do not take estrogens. Raloxifene, a selective estrogen receptor modulator has estrogen-like effects on bone tissue, and antagonize the action of estrogens on endometrium and breast tissue. It is unknown if the treatment of osteoporosis with raloxifene has estrogen-like or opposite effects on back pain and functional capacity among postmenopausal women with osteoporosis. A total of 120 postmenopausal women with osteoporosis and chronic back pain were randomized to receive raloxifene 60 mg with 1,000 mg calcium, and 800 IU vitamin D daily or 1,000 mg calcium and 800 IU vitamin D daily. Pain intensity and pain-related disability were measured before treatment at 6 months and after 1 year. Repeated measures of ANOVA, did not reveal statistically significant differences over time, on pain intensity and disability scores, between groups studied. There was a trend in pain intensity changes during the follow-up period, but the differences between the groups were not statistically significant. It seems that treatment with raloxifene does not influence back pain and disability among postmenopausal women with osteoporosis. Raloxifene may have estrogenic agonist effects on nociceptive processing in the central nervous system. PMID- 15834594 TI - Bibliometric analysis of global trends for research productivity in microbiology. AB - In order to expand upon the limited literature estimating the quantity and quality of worldwide research production in the field of microbiology, a bibliometric analysis was conducted for the period 1995-2003 using the PubMed and Journal Citation Reports databases. By searching the "microbiology" category of the Journal Citation Reports database, a total of 74 journals were identified that were also included in PubMed. From these journals, a total of 89,527 articles were identified for analysis, and data on the country in which the research originated was available for 88,456 (98.8%) of them. The individual countries were separated into nine world regions. In terms of research production for the period studied, Western Europe exceeded all other world regions, with the USA ranking second. The mean impact factor was highest for the USA at 3.4, while it was 2.8 for Western Europe and 2.4 for the rest of the world combined. The research productivity per unit of expenditure for research and development was higher for Canada and Western Europe than for the USA. The three regions in which research productivity increased the most were Asia, Latin America, and Eastern Europe. PMID- 15834595 TI - A cartilage growth mixture model for infinitesimal strains: solutions of boundary value problems related to in vitro growth experiments. AB - A cartilage growth mixture (CGM) model is linearized for infinitesimal elastic and growth strains. Parametric studies for equilibrium and nonequilibrium boundary-value problems representing the in vitro growth of cylindrical cartilage constructs are solved. The results show that the CGM model is capable of describing the main biomechanical features of cartilage growth. The solutions to the equilibrium problems reveal that tissue composition, constituent pre stresses, and geometry depend on collagen remodeling activity, growth symmetry, and differential growth. Also, nonhomogeneous growth leads to nonhomogeneous tissue composition and constituent pre-stresses. The solution to the nonequilibrium problem reveals that the tissue is nearly in equilibrium at all time points. The results suggest that the CGM model may be used in the design of tissue engineered cartilage constructs for the repair of cartilage defects; for example, to predict how dynamic mechanical loading affects the development of nonuniform properties during in vitro growth. Furthermore, the results lay the foundation for future analyses with nonlinear models that are needed to develop realistic models of cartilage growth. PMID- 15834596 TI - Arthrobacter ardleyensis sp. nov., isolated from Antarctic lake sediment and deep sea sediment. AB - Three psychrotrophic Arthrobacter strains, isolated from Antarctic lake sediment (An24, An25T) and deep-sea sediment (ZX6) were studied. Their 16S rRNA gene sequences showed highest similarities (97.0-97.9%) with those of A. nicotianae and A. protophormiae. All three strains underwent rod-coccus morphological change, had high mol% G+C content, were aerobic to slightly anaerobic, and grew between 0 degrees C and 30 degrees C, with optimal growth temperature around 25 degrees C. The cell wall peptidoglycan was A4alpha variant. DNA-DNA hybridization, physiological and chemotaxonomic studies indicated that these three strains constituted a new homogeneous genomic species within the genus Arthrobacter, for which the name Arthrobacter ardleyensis, with the type strain An25T (CGMCC 1.3685, JCM 12921) was proposed. PMID- 15834597 TI - Substituent effects on the oxidation of substituted biphenyl congeners by type II methanotroph strain CSC1. AB - The oxidation potential of type II groundwater methanotroph, strain CSC1, expressing soluble methane monooxygenase, was measured in the presence of 10 ortho-substituted biphenyls with varying electronics, sterics, and hydrophobicity character for comparison with type II Methylosinus trichosporium OB3b. Strain CSC1 showed faster rates with all compounds tested, with the exception of 2 nitrobiphenyl, 2-hydroxybiphenyl, and 2-aminobiphenyl. Products of oxidation observed upon incubation of strain CSC1 with biphenyl and 2-hydroxybiphenyl were hydroxylated biphenyls that revealed less preference for the para position and different dihydroxylation positions, respectively, in comparison to those observed with M. trichosporium OB3b. Only the intramolecular hydrogen migration, or NIH-shift, product was observed in the case of 2-chlorobiphenyl, whereas M. trichosporium OB3b yielded a variety of chlorohydroxybiphenyls. Quantitative structure-biodegradation relationships constructed with the maximum observed oxygen uptake rates as a dependent variable and a variety of descriptors showed an influence of substituent electronic character on the oxidation activity of strain CSC1. However, compound hydrophobicity and not compound size, as was observed with M. trichosporium OB3b, was shown to influence rates to a greater extent. This suggests that transport of the compound through the cell membrane and to the sMMO active site is rate-determining for strain CSC1. PMID- 15834598 TI - [Toxic contact dermatitis from poison ivy in a private garden in Germany]. AB - A couple suffered for 5 years from recurrent eruptions with vesicles and bullae after contact with an unknown "climbing weed" in their private garden in Germany. After this plant was identified as poison ivy and eradicated, their skin problems were solved. This is the first report of poison ivy in this setting. Urushiols in poison ivy are not only strong allergens but also potent irritants. Negative patch tests in the husband suggest that the bullous dermatitis was a toxic reaction. PMID- 15834599 TI - Visual acuity and structural findings in old age-related macular degeneration. AB - BACKGROUND: Although exudative age-related macular degeneration (AMD) leads to a substantial visual loss in most patients there is still significant variation in the end- stage visual acuity level. We analysed lesions in eyes with long standing AMD in order to find contributing factors for this variation. METHODS: Sixty-one out of 121 patients examined for exudative AMD and still alive 4.8-9.2 (mean 6.8) years after the acute phase were re-examined. The lesion size, area of subretinal fibrosis, geographic atrophy, presence of a persistent exudative process, and shortest distance to normal looking retina were measured from digital fundus photographs taken at the re-examination and correlated with visual acuity. RESULTS: Lesion size, the presence of a continuing exudative process, or subretinal fibrosis were independent predictors for poor vision. Better vision in the other eye was connected with poor vision in the affected study eye. CONCLUSIONS: In addition to lesion size, the presence of a continuing exudative process and subretinal fibrosis also have deleterious effects on long-term visual acuity after exudative AMD. PMID- 15834600 TI - Trombophilic screening for nonarteritic anterior ischemic optic neuropathy. AB - BACKGROUND: Nonarteritic anterior ischemic optic neuropathy (NAION) is an ischemic infarction of the optic nerve head, frequently leading to sudden, mostly irreversible loss of vision. In this study blood thrombophilic factors, as well as cardiovascular risk factors were investigated for their relevance to this pathology. Trombophilic risk factors so far not evaluated were included in the study. PATIENTS AND METHODS: 37 NAION patients (4 with sequential second eye involvement) and 81 matched control subjects were examined. From blood, protein C, protein S, antithrombin, von Willebrand antigen levels (vWFAg), and factor V (Leiden) mutation, factor VIIIC level, plasminogen activity, lipoprotein (a) and fibrinogen levels, and presence of anticardiolipin antibodies were investigated. Possibly relevant pathologies [e.g. diabetes mellitus (DM), hypertension, and ischemic heart disease] were also registered. RESULTS: Elevated Lp(a) and vWFAg levels, DM, F V (Leiden), hypercholesterolemia, and hyperfibinogenemia proved to be significant risk factors associated with NAION. Forward stepwise logistic regression analysis revealed that high Lp(a), DM, and FV (Leiden) were the main predictive components, with odds ratios 16.88 (p=0.012), 5.78 (p=0.022) and 4.44 (p=0.033), respectively. CONCLUSIONS: Based on our results it appears that thrombophilia is likely to contribute to the development of NAION besides vascular damage due to the presence of cardiovascular risk factors. Further data are needed, however, to justify the suggested use of secondary prophylaxis using anticoagulant/antiplatelet therapy. PMID- 15834601 TI - Asymmetric pigmentary glaucoma in a patient with Marfan's syndrome. AB - BACKGROUND: No relationship between Marfan's syndrome and pigmentary glaucoma has previously been described in the ophthalmic literature. We describe the case of a patient with Marfan's syndrome who had bilateral pigment dispersion syndrome and asymmetrical glaucoma. METHODS: A 34-year-old man with Marfan's syndrome developed pigment dispersion bilaterally. In the right eye, elevated intra-ocular pressure was associated with marked glaucomatous excavation of the right disc and corresponding visual field loss. A localised zonular dehiscence was present at the 6 o'clock position in the right eye. RESULTS: Ultrasound biomicroscopy showed marked iris concavity in the right eye, increased iridolenticular contact and superior subluxation of the right lens, resulting in increased dispersion of pigment unilaterally. This explains the asymmetric pattern of the glaucomatous damage. CONCLUSIONS: Asymmetrical progression of pigmentary glaucoma is uncommon and should prompt a search for some other mechanical factor, which might increase the iridozonular contact on one side or reduce it on the other. The association between the two syndromes in this patient was most likely mechanical due to reduced fibrillin expression throughout the eye and particularly in the iris and the lens zonules, resulting in loss of iris tensile strength and marked iris concavity as well as zonular weakness and partial lens subluxation. PMID- 15834602 TI - Discrimination between normal and glaucomatous eyes using Stratus optical coherence tomography in Taiwan Chinese subjects. AB - BACKGROUND: We differentiated between normal and glaucomatous eyes in the Taiwan Chinese population based solely on the quantitative assessment of summary data reports from Stratus optical coherence tomography (OCT) by comparing their area under the receiver operating characteristic (ROC) curve. METHODS: One randomly selected eye from each of the 62 patients with early glaucomatous damage (mean deviation -2.8 +/- 1.8 dB) and from each of the 62 age- and sex-matched normal individuals were included in the study. Measurements of glaucoma variables (retinal nerve fiber layer thickness and optic nerve head analysis results) were obtained by Stratus OCT. Twenty-one OCT parameters were included in a linear discriminant analysis (LDA) using forward selection and backward elimination to determine the best combination of parameters for discriminating between glaucomatous and healthy eyes based on ROC curve area. RESULTS: The average RNFL thickness was the best individual parameter for differentiating between normal eyes and glaucomatous eyes (ROC curve area 0.793). The maximum area under the ROC curve of six input parameters (average RNFL thickness; 10, 11, and 12 o'clock segment thicknesses; cup area; and vertical integrated rim area) generated by the forward selection method was 0.881. Whereas the maximum area under the ROC curve of 15 input parameters (average RNFL thickness; 1, 3, 4, 6, 8-10, 12 o'clock segment thicknesses; vertical integrated rim area; horizontal integrated rim area; disc area; cup to disc area ratio; cup to disc horizontal ratio; and cup to disc vertical ratio) generated by backward elimination method was 0.929. CONCLUSIONS: The performance of individual parameters obtained from Stratus OCT is fairly reliable for differentiating the early glaucomatous eyes from normal eyes. However, the discriminant power increases when LDA with forward selection and backward elimination methods is applied. PMID- 15834603 TI - Normal values for Octopus tendency oriented perimetry in children 7 through 13 years old. AB - BACKGROUND: The purpose of this study was to determine age-dependent values for mean sensitivity, mean deviation, and loss variance for normal children age 6 through 13 years, using the Octopus 301 perimeter and the Tendency Oriented Perimetry 32 (TOP-32) program. METHODS: Healthy children from 6 through 13 years of age with a visual acuity of 20/20 OU and at least 60 arc seconds of stereopsis were recruited. They were tested on the Octopus 301 perimeter using the TOP-32 program, and each eye was tested twice during one session. Results for all four tests were averaged for each subject, and the average was used for statistical analysis. The main outcome measures were mean sensitivity, mean deviation, and loss variance by age. The test duration, learning and fatigue effects, and the influence of false positive responses on the average mean sensitivity were also analyzed. RESULTS: There were 142 subjects tested. Six-year-old children showed high intersubject variability and were excluded (N=23). The average age of the remaining 7- to 13-year-old cohort (N=119) was 9.8 +/- 1.7 years. The average test duration was 2.9 +/- 0.3 min. The average mean sensitivity was 28.7 +/- 1.9 dB. The mean sensitivities for tests 1 through 4 were 28.14, 28.63, 28.96, and 28.92 dB, respectively. The average mean deviation was 0.4 +/- 1.9 dB. The slope of the regression line for mean sensitivity vs age was -0.018 +/- 0.165 dB/year, which was not significantly different from zero (two-tailed t test, p=0.83). The false positive catch trial rate was high (mean 26%) and was independent of age. When the data from subjects who had more than two false positive catch trial responses on any one test were eliminated, the mean sensitivity decreased to 28.3 +/- 1.9 dB. CONCLUSIONS: When testing patients age 7 through 13 years on the Octopus 301 perimeter using the TOP-32 program, comparison against the programmed normal mean sensitivity value for 20-year-old subjects (29.0 dB) is appropriate. During a sequence of four tests, both learning and fatigue effects are evident. The false positive response rate is naturally high regardless of age, and children should not be overencouraged to respond during testing. PMID- 15834604 TI - Tear film function and corneal sensation in the early postoperative period after LASEK for the correction of myopia. AB - PURPOSE: Subepithelial nerves and stromal nerves of the cornea are damaged during LASEK surgery for the correction of myopia. This leads to a reduction in corneal sensation and to alterations of the tear film function in the early postoperative period. The aim of this study was to evaluate tear film function, corneal sensation and subjective symptoms of dry eye in the early postoperative period after LASEK for the correction of myopia. METHODS: LASEK surgery was performed in 20 eyes of 10 consecutive patients for the correction of myopia (-1.0 to -9.0 D, mean -4.86 D). Subjective symptoms of a dry eye were evaluated with a standardised questionnaire. Schirmer test with and without local anaesthesia, tear film break-up time, fluorescein staining of the cornea and corneal esthesiometry (Cochet-Bonnet) were performed before surgery and 1 week, 1 month, 2 months and 3 months after LASEK. Additionally corneal staining and subjective symptom severity scores were assessed 3 days after surgery. RESULTS: Corneal sensation was reduced 1 week after LASEK and recovered during the first month after LASEK. The tear film break-up time was reduced 1 week and 1 month after LASEK and reached preoperative values 2 months after surgery. Fluorescein staining was increased 3 days and 1 week after LASEK. Subjective symptom severity scores were increased 3 days, 1 week, 1 month and 2 months after LASEK. Schirmer tests values with local anaesthesia were significantly reduced at 3 months after surgery, but not at 3 days, 1 month, or 2 months. Schirmer test values without local anaesthesia were significantly decreased 2 and 3 months after LASEK, but not after 3 days and 1 month after LASEK. CONCLUSIONS: LASEK alters ocular surface haemostasis and reduces corneal sensation in the early postoperative period. Subjective symptoms of dry eye were described during the first 2 months after surgery. PMID- 15834605 TI - 'Xerosis meter': a new concept in dry eye evaluation. AB - BACKGROUND: Dry eye is not only incapacitating to the patient but its treatment is also challenging. It would undoubtedly be more amenable to therapy if it could be detected at an early stage and its prognosis be recorded accurately and sensitively. In the past few years 'dry eye' and its sequelae have become the focus of attention of ophthalmologists worldwide. Whereas there has been a tremendous contribution by the pharmaceutical industry towards its treatment, its diagnostic and prognostic tests, such as Schirmer's test and tear film break-up time (BUT), appear primitive. With this in mind, we have designed a 'xerosis meter'--an electronic device that can detect and grade tissue dryness. METHODS: This device is based on the principle that the electrical conductivity of any tissue is directly proportional to its wetness. The sensitivity of this instrument was compared with Schirmer's test and BUT. RESULT AND CONCLUSION: The xerosis meter readings in normal eyes (control group) and dry eyes (test group) were compared statistically using the unpaired t-test (p<0.001). The sensitivity of the xerosis meter (86.11%) was much higher than that of Schirmer's test (80.55%) and BUT (66.66%). PMID- 15834606 TI - Toxic effects of indocyanine green, infracyanine green, and trypan blue on the human retinal pigmented epithelium. AB - BACKGROUND: Indocyanine green, infracyanine green, and trypan blue are frequently used as aids to visualize structures removed during vitreoretinal surgery. But they may have toxic effects on the retina. We therefore compared the acute and chronic toxicities of these stains on cultured human retinal pigmented epithelial (RPE) cells using clinically relevant concentrations and an identical experimental setup for each agent. METHODS: Monolayers of RPE cells were incubated with various concentrations of indocyanine green, infracyanine green (each at 0.005%, 0.05%, and 0.5%) or trypan blue (0.05%, 0.06%, 0.1%, 0.15%, and 0.5%) for 5 min (acute exposure) or 6 days (chronic exposure). Using the propidium iodide assay, acute cytotoxicity was monitored at 15-min intervals for up to 3 h. Chronic cytotoxicity was assessed by monitoring cell calcein esterase activity, cell proliferation, and cell morphology (viability) after 6 days of exposure. RESULTS: Indocyanine and infracyanine green induced acute and chronic toxicities at a concentration above 0.05%. Trypan blue evoked no acute toxicity, but it was chronically cytotoxic at all tested concentrations. CONCLUSIONS: Despite thorough rinsing after application, significant amounts of the not sufficiently water soluble indocyanine and infracyanine green are retained after surgery by the eye. Trypan blue, being more water-soluble than ICG, is probably retained to the least degree. This circumstance is fortunate given that trypan blue exhibits a chronic cytotoxicity comparable to ICG at all clinically relevant concentrations. During vitrectomy, surgeons should aim to expose retinal tissue to only low concentrations of these stains and for as short a period as possible. PMID- 15834607 TI - Binocular fixation topography in patients with diabetic macular oedema: possible implications for photocoagulation therapy (3rd revision). AB - BACKGROUND: During retinal photocoagulation for diabetic maculopathy, there is a potential risk of foveal burns, and laser scars may later enlarge to be sight threatening when involving retinal areas previously used during fixation. Since the retinal area used during binocular steady fixation has been found to vary considerably in the normal test person and central fixation may be even further compromised in patients with diabetic maculopathy, the sight-threatening side effects could possibly be reduced by taking into account the fixation area individually. However, no study has described and quantified the retinal area of fixation binocularly in patients with clinically significant macular oedema (CSME). METHODS: Sixteen diabetic patients with CSME in one or both eyes were examined. Each examination included visual acuity testing (ETDRS charts), a standard eye examination, central retinal thickness assessment by optical coherent tomography, fluorescein angiography and binocular quantification of fixational eye movements using an infrared recording technique. RESULTS: A negative correlation was found between visual acuity and mean microsaccadic amplitude (R=0.48, p=0.009). The maximal retinal extension of the fixation area ranged between 1.0 degrees and 3.0 degrees , and in two eyes with CSME, this area was estimated to exceed 800 mum on the retinal plane. No correlation was found between retinal thickness and visual acuity, retinal area of fixation, maximal extension of the fixation area or mean microsaccadic amplitude. CONCLUSION: Large interindividual differences in quantitative measures of binocular fixational eye movements were found. The mean amplitude of fixational eye movements was not correlated to central retinal thickness, and fixation area could only partly be predicted by visual acuity. Two eyes with CSME had an estimated maximal extension of the fixation area exceeding the central 800 mum on the retinal plane; thus, the possible benefit of individualising central photocoagulation according to precise measures of fixation area needs to be investigated on a larger population. PMID- 15834608 TI - Photodynamic therapy for wound-healing modulation in pterygium surgery. A clinical pilot study. AB - BACKGROUND: This study was performed to evaluate the safety and efficacy of photodynamic therapy with the carboxyfluorescein ester BCECF-AM as an adjunctive treatment procedure for pterygium surgery to reduce the rate of recurrence. METHODS: In this nonrandomized prospective clinical trial, 19 eyes with nasally located primary pterygium were examined. All eyes were treated with the bare sclera surgical technique. Seven eyes received in addition treatment with BCECF AM solution and blue light. All patients were evaluated at least after 1 day, 1 week, 1 month, 3 months, and 1 year. Postoperative fibrovascular growth from the limbus of at least 1 mm was defined as recurrence. RESULTS: The intraoperative application of BCECF-AM solution did not cause anterior chamber flare or any other significant side effects. The bare sclera surgery rate of recurrence was 0% (zero of 12) after 3 months and 91% (11 of 12) after 1 year. The additional photodynamic therapy treatment had a rate of recurrence of 14.2% (one of seven) after 3 months and 71.4% (five of seven) after 1 year. CONCLUSIONS: The applied PDT technique seems to be a safe procedure but is associated with a high rate of recurrence. In conclusion, the evaluated PDT treatment procedure, at this point, should not be considered. As we found a high rate of recurrence also in the control group, the bare sclera technique is not effective, even in primary pterygia. PMID- 15834609 TI - Effect of alpha2-macroglobulin on retinal glial cell proliferation. AB - BACKGROUND: Activation of the receptor for alpha2-macroglobulin (alpha2 M), the low-density lipoprotein-related protein (LRP1; CD91), has been suggested to represent a possible strategy for the inhibition of uncontrolled retinal cell proliferation via stimulation of the clearance of alpha2 M-bound growth factors and proteinases from the extracellular space. In order to prove this assumption, we investigated the effect of alpha2 M on the proliferation of Muller glial cells in vitro. METHODS: Proliferation assays using bromodeoxyuridine were carried out on cultured Muller glial cells of the guinea pig in the absence and presence of alpha2 M. RESULTS: Activated alpha2 M evoked a slight increase of the cell proliferation at control conditions. Addition of alpha2 M to the culture medium inhibited the proliferation evoked by agonists of G-protein-coupled receptors [adenosine 5'-triphosphate (ATP), neuropeptide Y]. However, alpha2 M did not diminish the proliferation evoked by agonists of receptor tyrosine kinases (epidermal and platelet-derived growth factors) and by serum, respectively. Inhibition of LRP1 by a neutralizing antibody did not alter the ATP-evoked proliferation while it increased the proliferation in the presence of alpha2 M. CONCLUSION: It is concluded that alpha2 M inhibits the proliferation evoked by agonists of G-protein-coupled receptors, possibly via enhanced growth factor clearance by LRP. PMID- 15834610 TI - A microbiological and confocal microscopy study documenting a slime-producing Staphylococcus epidermidis isolated from a nylon corneal suture of a patient with antibiotic-resistant endophthalmitis. AB - BACKGROUND: We describe a case of posttraumatic endophthalmitis unresponsive to systemic (amoxicillin+clavulanic acid and piperacillin/tazobactam), intra-ocular (vancomycin) and topical (ofloxacin, tetracycline and sulfametoxazole) antibiotic therapy. Microbiological and confocal microscopy studies of a nylon corneal suture revealed the presence of a slime-producing strain of Staphylococcus epidermidis. METHODS: We describe the history and clinical presentation of a 77 year-old man in whom a high-grade posttraumatic endophthalmitis resolved only after the removal of a single nylon corneal suture. Microbiological investigations of the aqueous, vitreous and suture were performed, and the propensity of the suture-associated isolate to form biofilm was assessed using confocal microscopy. RESULTS: A single stain of S. epidermidis was isolated from both aqueous and vitreous specimens and from the suture. The planktonic form of the isolate was susceptible in vitro to the antibiotics administered to the patient, but the strain was capable of forming biofilms and this phenotype showed resistance to high concentrations of the same antibiotics. CONCLUSIONS: The presence of a slime-producing strain of S. epidermidis should be considered in endophthalmitis that is unresponsive to specific antibiotic therapy, especially in cases in which an intra-ocular foreign body (e.g., a suture) is present. PMID- 15834611 TI - Autoantibodies in patients with glaucoma: a comparison of IgG serum antibodies against retinal, optic nerve, and optic nerve head antigens. AB - PURPOSE: The aim of this study was to analyze and compare the entire IgG autoantibody patterns against different ocular antigens (retina, optic nerve, and optic nerve head) in sera of glaucoma patients and healthy subjects. METHODS: Sixty-six patients were included in this study: healthy volunteers without any ocular disorders (CO, n=30), patients with primary open-angle glaucoma (POAG, n=19), and patients with normal-tension glaucoma (NTG, n=17). The sera were tested for antibodies against retinal, optic nerve, and optic nerve head tissues. Immunodetection was performed using 4-chloro-1-naphthol staining. The autoantibody patterns were digitized and subsequently analyzed by multivariate statistical techniques. RESULTS: All patients showed a complex repertoire of IgG antibodies against retinal, optic nerve, and optic nerve head antigens. The analysis of discriminance revealed a statistically significant differences between the patterns of all three groups. Our multivariate approach could quantify the differences in immunoreactivities of patient sera against the three antigens. The POAG group had the most significant difference against retinal antigens (P=0.0021) compared with the other antigens. In the NTG group the highest reactivity appeared against optic nerve head (P=0.00053) and optic nerve (P=0.0025). CONCLUSIONS: All groups showed different and complex antibody patterns against the three ocular tissues. These autoantibodies are highly specific for each patient group. The analysis of these patterns could provide further information about possible autoimmune mechanisms in the pathogenesis of glaucoma. PMID- 15834612 TI - Double frameshift mutations in APC and MSH2 in the same individual. AB - Heterozygous germline DNA mismatch repair gene mutations are typically associated with HNPCC. Here we report the case of a proband whose father was known for familial adenomatous polyposis. The number of polyps (less than ten) was not typical of polyposis; therefore, the diagnosis of HNPCC was entertained. Microsatellite instability analyses were performed on peripheral blood and biopsy of a right-sided dysplastic adenoma. The tumour tissue showed high-grade instability, and subsequently, immunohistochemistry showed that neither MSH2 nor MSH6 proteins were expressed in tumour cells. Prophylactic colectomy was performed, and an adenocarcinoma developing within the adenoma was diagnosed (pT1N0). Genomic DNA analysis revealed a novel mutation in MSH2 as a frameshift mutation in exon 7 (c.1,191_1,192dupG). Both parents of the proband were analyzed for MSH2 and APC mutations, and in the father, a truncating mutation in exon 15 of APC was identified as del3471-3473GAGA. This mutation was found to be present in the proband. His mother was found to bear the MSH2 exon 7 mutation. At follow up, the proband was diagnosed with fundic, antral and duodenal adenomas (one fundic adenoma showed low-grade dysplasia). Several tubular rectal adenomas with low-grade dysplasia were excised. The patient later developed an intra-abdominal desmoid tumour. PMID- 15834613 TI - Adeno-associated virus vectors are able to restore fatty aldehyde dehydrogenase deficiency. Implications for gene therapy in Sjogren-Larsson syndrome. AB - Sjogren-Larsson Syndrome (SLS) is caused by an autosomal recessive defect in the gene coding for fatty aldehyde dehydrogenase (FALDH), an enzyme necessary for the oxidation of long-chain aliphatic aldehydes to fatty acid as one enzyme of the fatty alcohol:nicotinamide-adenine dinucleotide (NAD+)-oxidoreductase complex (FAO). The impaired activity of FALDH leads to the clinical symptom triad of generalized ichthyosis, mental retardation, and spastic diplegia or tetraplegia. Treatment options are primarily symptomatic. Gene therapy by means of genetic reintroduction of the functional FALDH gene into defective cells has so far not been considered as a therapeutic modality. In order to pursue such an approach for SLS, we constructed a recombinant adeno-associated virus-2 vector containing the human cDNA of functional FALDH and evaluated its capability to restore the enzyme-deficiency in a FALDH-deficient cell line resembling the gene defect of SLS. rAAV-2 transduction of FALDH-deficient cells, usually exhibiting less than 10% of normal FALDH activity, resulted in an increase of FALDH activity within the range of unaffected cells. Moreover, FALDH-transduced cells regained resistance over exposure to long chain aldehydes, which are otherwise toxic to FALDH-deficient cells. These results indicated that rAAV-2 vectors are able to restore FALDH-deficiency in a cell system resembling SLS. The findings give the first support to the concept that gene therapy might be a future option for the treatment of SLS. PMID- 15834614 TI - Percutaneous penetration through slightly damaged skin. AB - Guidelines for experimental studies of percutaneous penetration prescribe optimal barrier integrity of the skin. The barrier integrity of the skin exposed in occupational or household situations is, however, not always ideal, and skin problems are among the most dominant reasons for absence from work. We have therefore evaluated an experimental model for percutaneous penetration through slightly damaged skin. The influence of a slight damage to the skin was evaluated using five pesticides covering a wide range of solubilities. We used an experimental model with static diffusion cells mounted with human skin. A slight damage to the barrier integrity was induced by pre-treatment of the skin with sodium lauryl sulphate (SLS) before pesticide exposure. The experimental model with 3 h pre-treatment with SLS (0.1% or 0.3%) assured a significant but controlled damage to the barrier integrity, a damage that remained unchanged for an experimental period of 48 h. Based on the percutaneous penetration of five pesticides, we conclude that a slightly damaged skin may significantly affect the rate, lag-time as well as total penetration of chemicals covering a wide range of solubilities. The percutaneous penetration of the most hydrophilic compounds will be those most affected. These findings should be considered when setting standards for dermal exposure to chemicals. PMID- 15834615 TI - Contribution of manganese peroxidase and laccase to dye decoloration by Trametes versicolor. AB - During dye decoloration by Trametes versicolor ATCC 20869 in modified Kirk's medium, manganese peroxidase (MnP) and laccase were produced, but not lignin peroxidase, cellobiose dehydrogenase or manganese-independent peroxidase. Purified MnP decolorized azo dyes [amaranth, reactive black 5 (RB5) and Cibacron brilliant yellow] in Mn(2+)-dependent reactions but did not decolorize an anthraquinone dye [Remazol brilliant blue R (RBBR)]. However, the purified laccase decolorized RBBR five to ten times faster than the azo dyes and the addition of a redox mediator, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), did not alter decoloration rates. Amaranth and RB5 were decolorized the most rapidly by MnP since they have a hydroxyl group in an ortho position and a sulfonate group in the meta position relative to the azo bond. During a typical batch decoloration with the fungal culture, the ratio of laccase:MnP was 10:1 to 20:1 (based on enzyme activity) and increased to greater than 30:1 after decoloration was complete. Since MnP decolorized amaranth about 30 times more rapidly than laccase per unit of enzyme activity, MnP should have contributed more to decoloration than laccase in batch cultures. PMID- 15834616 TI - Light-dependent PER-like proteins in the cephalic ganglia of an apterygote and a pterygote insect species. AB - Antibodies targeted to a highly conserved tetradecapeptide region of the pivotal biological clock protein PER detect in the firebrat Thermobia domestica a 115-kDa protein and in the cockroach Periplaneta americana a 110-kDa protein that are present in the cytoplasm of a small set of brain cells. A similar cytoplasmic reaction occurs with antisera to the whole PER protein of Drosophila melanogaster, but these antisera also react with numerous cell nuclei. On western blots, they detect an 80-kDa antigen in T. domestica and 70- and 80-kDa antigens in P. americana. No indication of antigen translocation between cell nuclei and cytoplasm was found. Nuclear staining is maintained at a high constant level in T. domestica held at a 12:12 h light:dark photoperiod (LD) or in continuous light, but disappears rapidly in response to extended darkness. In P. americana under LD conditions, the number of immunoreactive nuclei and their staining intensity fluctuate in parallel, with maximal staining late in the day. The circadian changes are maintained in continuous light but all staining vanishes in continuous darkness. A 6-h light pulse in early night of an LD cycle induces maximal staining after about 10 h, suggesting that the effect of light on nuclear PER-like expression is indirect. The behaviour of nuclear antigens is opposite to that of the cytoplasmic PER-like proteins that persist in constant darkness and disappear in constant light. Under LD conditions, the cytoplasmic PER-like antigen cycles in T. domestica but remains at a steady level in P. americana. The sensitivity to photoregime suggests that both the nuclear and the cytoplasmic PER like antigens are components of the biological clock. PMID- 15834617 TI - [Subjective aspects in mothers of speech impaired children]. AB - OBJECTIVE: The purpose of this study was to evaluate the subjective burden in mothers of speech impaired children against the presence or absence of emotional disorders such as anxiety and depression. METHODS: A total of 89 mothers (age 33.3+/-5.5 years, range: 19.6-43.8 years) of 89 speech impaired preschool children (23 girls, 66 boys, age: 3.9+/-1.5 years, range: 1.6-7.1 years) were examined. Screening for a depressive or anxiety disorder was performed using the German version of the Hospital Anxiety and Depression Scale HADS-D. The "Short Questionnaire on Current Burden" (Kurzfragebogen zur Aktuellen Belastung, KAB) was used to measure the level of perceived subjective burden at the time of the investigation. RESULTS: A higher score in the KAB test occurred significantly more often in mothers with a positive HADS depression/anxiety subscale result than in those with a negative one. CONCLUSIONS: There is a need for specific assessment of wellbeing in mothers of speech impaired children. In clinical practice, one may restrict oneself to the use of a screening test for anxiety and depression. PMID- 15834618 TI - Long-term outcomes of children with end-stage renal disease. AB - Long-term survival of children with end-stage renal disease (ESRD) has increased in the last 20 years, but the mortality rate remains high. Cardiovascular disease accounts for 40 to 50% of all deaths, infectious disease for about 20%. A prolonged period of dialysis versus having a renal graft and persistent hypertension are mortality risk factors. The prevalence of the various morbidities is high among those who have reached adulthood. Nearly 50% of all these patients suffer from left ventricular hypertrophy and life-threatening vascular changes; nearly one third has clinical signs of metabolic bone disease. This accounts for both dialysis and transplant recipients. The chance of getting cancer is increased ten times compared to the general population; skin cancer and non-Hodgkin lymphomas are most commonly reported. A long period of dialysis at childhood is associated with impairment of both cognitive and educational attainment. However, despite all these negative outcomes, the health perception of young adults with childhood onset ESRD is positive. Research and therapy in children with ESRD should focus not only on prevention of graft failure, but also on prevention of co-morbidity, especially cardiovascular disease, life threatening infections and malignancies. Early transplantation, more extended forms of frequent hemodialysis in those who can not be transplanted, a more rigorous treatment of hypertension, avoidance or at least dosage reduction of calcium-containing phosphate binders, reduction of the chronic inflammatory state, and tailor made anti-rejection therapy after transplantation may all be targets to improve the outcome in future patients. PMID- 15834619 TI - Prospective surveillance of Canadian children with the haemolytic uraemic syndrome. AB - We determined prospectively the incidence of childhood diarrhoea associated haemolytic uraemic syndrome (D+ HUS) or HUS due to Streptococcus pneumoniae (SP+ HUS) in Canada through the Canadian Paediatric Surveillance Program (CPSP) network. From April 2000, to March 2002, 82% (1,886/2,300) of all Canadian paediatricians reported possible new cases of D+ HUS or SP+ HUS. Diagnosis was validated with a second questionnaire. There were a total of 136 children with D+ HUS (epidemic, n =15; endemic, n =121), corresponding to an annual incidence rate of 1.11 cases per 100,000 children under 16 years of age. Excluding incomplete forms, the annual incidence rate among endemic cases was 0.74 cases per 100,000. The provinces of Ontario (40%), Quebec (31%), and Alberta (18%) accounted for 89% of cases. Evidence of E. coli O157 or O157:H7 were found in 94% (79/84) of children who had positive stool cultures. The mortality rate was 4% (n =5) and 34% (n =41) of children underwent dialysis for a median of 12 days (2-60 days). SP-HUS was diagnosed in four children with pneumonia and pleural effusion (n =2) or meningitis (n =2) who survived. One child with positive direct Coombs testing had definitive evidence of SP+ HUS. The remaining three were considered possible cases. We conclude that SP+ HUS is rare in Canada. Over the last 15 years, the incidence of childhood D+ HUS in Canadian children may have decreased. PMID- 15834620 TI - Parathormone sensitivity and responses to protein kinases in subclones of opossum kidney cells. AB - Parathyroid hormone (PTH) inhibits sodium-dependent phosphate (Na(+)-Pi) transport in the renal proximal tubule and opossum kidney (OK) cells by mechanisms involving protein kinases (PK) A and C, and 20-hydroxyeicosatetraneoic acid (20-HETE). The magnitude of the effect of PKA and PKC on Na(+)-Pi transport in OK cells varies in different studies, suggesting that OK cell subclones are functionally heterogeneous despite their morphological similarity. We studied the effect of PTH and PK effectors in two separate sets of OK cells at two different time periods. Each group of cells were derived from the same stock, at passages 75-85. In one group of OK cells 20-HETE (10(-7 )M) induced a 24% decrease in Na (32)Pi transport. Addition of PTH (10(-7) M) inhibited Pi transport by 44%. Addition of TPA (10(-8) M) resulted in a 32% decrease in Na-(32)Pi transport. Exposure of cells to the PKC inhibitor staurosporine (10(-7) M) induced a significant increase in Na-(32)Pi transport. Simultaneous addition of 20-HETE and staurosporine restored baseline Pi transport. Finally, Br-cAMP (10(-7) M) inhibited Na-(32)Pi transport by 32%. In another group of OK cells we reexamined the affect of these substances on Na-(32)Pi transport. 20-HETE (10(-7) M) induced a significant increase (30%) in Na-(32)Pi transport. PTH (10(-7) M) had no effect on Na-(32)Pi transport (P = 0.05). TPA (10(-8) M) induced a 42% increase in Na (32)Pi transport (P < 0.01). Staurosporine (10(-7) M) induced a slight decrease in Na-(32)Pi transport (P < 0.05). Simultaneous addition of 20-HETE and staurosporine restored Na-(32)Pi transport to baseline levels. Finally, Br-cAMP (10(-7) M) inhibited Na-(32)Pi transport by 23%. We conclude that different groups OK cells have markedly different responses to regulators of Na-Pi cotransport. PMID- 15834621 TI - Determination of inulin clearance by single injection or infusion in children. AB - The reference method to determine the glomerular filtration rate (GFR) in children is the urinary clearance of inulin during a continuous intravenous infusion. Alternatively, the plasma clearance of inulin can be determined, which does not require urine collection. This study compared the determination of the inulin plasma clearance in 24 pediatric patients by two methods: the single injection and the continuous infusion method. In the single injection method 5000 mg/m(2) inulin was administered as bolus injection, and blood samples were drawn 10, 30, 90, and 240 min after administration. For the continuous infusion method inulin was started overnight and blood samples were collected the next day. The inulin plasma clearance determined by the single injection method was on average 9.7 ml min(-1) 1.73 m(-2) higher than the clearance determined with the continuous infusion method (95% CI: 5.3-14.2). The difference between the two methods was smaller at lower GFRs. The difference in results generated by the two methods in children is small and is considered acceptable in clinical practice. For practical reasons, the single injection method with minimum sampling is preferred. PMID- 15834622 TI - Brain arteriovenous malformations: measurement of nidal volume using a combination of static and dynamic magnetic resonance angiography techniques. AB - Arteriovenous malformations of the brain are complex vascular lesions that are an important cause of death and long-term disability. Currently, catheter angiography (CA) is the reference standard procedure for the diagnosis and follow up of treated arteriovenous malformations (AVMs). This is an invasive procedure with potential risks. Magnetic resonance angiography (MRA) is commonly used in neurovascular imaging as a non-invasive alternative. Various MRA techniques have been used in the diagnosis and follow-up of AVMs but these have suffered from lack of temporal or spatial resolution. In this 60-patient study we describe the combination of two techniques: dynamic magnetic resonance digital subtraction angiography with a high temporal resolution, and a non-dynamic contrast-enhanced time-of-flight sequence with a high spatial resolution technique, in the assessment of AVM. The results showed an excellent correlation between MRA and CA measurement of both maximum linear dimension and AVM nidus volume. PMID- 15834623 TI - Use of a decision analysis model to assess the medicoeconomic implications of FDG PET imaging in diagnosing a solitary pulmonary nodule. AB - This study assessed the use of positron emission tomography (PET) in identifying and diagnosing solitary pulmonary nodules (SPNs). For this a decision analysis model was constructed, and three alternatives were compared: wait and watch (WW), PET and anatomical computed tomography (PET), and CT plus PET (CT+PET). Transition probabilities were estimated from published data and consultations with experts. Costs of diagnosis were derived from the French reimbursement scale, and treatment costs from a national hospital database of diagnosis-related groups. The base case was defined as a 65-year-old male smoker with a 2-cm SPN and an associated high risk of malignancy of 43%. Evaluation criteria included incremental cost-effectiveness ratios and the proportion of unnecessary operations avoided in patients without malignant SPN. For the base case WW was the least effective and cheapest strategy. CT+PET was more effective and presented lower incremental cost-effectiveness ratio (3,022 per life-year gained). It also was superior to PET in cost-effectiveness terms and resulted in 4.3% fewer unnecessary resections of benign SPN than did PET. Risk profile analyses performed on SPN malignancy risk showed that CT + PET remains the most cost-effective strategy in the range of 5.7-87%, and that WW is more cost effective in the range of 0.3-5.0%. CT+PET is thus cost-effective in detecting malignant SPN in patients with a risk of malignity of at least 5.7% and may avoid inappropriate resections of benign SPN. These findings support the attempts to introduce a larger number of PETs in France for SPN diagnosis. PMID- 15834624 TI - Threading neural feedforward into a mechanical spring: how biology exploits physics in limb control. AB - A solution is proposed of the hitherto unsolved problem as to how neural feedforward through inverse modelling and negative feedback realised by a mechanical spring can be combined to achieve a highly effective control of limb movement. The revised spring approach that we suggest does not require forward modelling and produces simulated data which are as close as possible to experimental human data. Control models based on peripheral sensing with forward modelling, which are favoured in the current literature, fail to create such data. Our approach suggests that current views on motor control and learning should be revisited. PMID- 15834629 TI - Sample size matters: a guide for surgeons. AB - Considerations of sample size computations in the medical literature have gained increasing importance over the past decade and are now often mandatory for scientific grant proposals, protocols, and publications. However, many surgeons are ill-prepared to understand the parameters on which the appropriate sample size is based. The present article has several objectives: first, to review the need for sample size considerations; second, to explain the ingredients necessary for sample size computations in simple, nonmathematic language; third, to provide options for reducing the sample size if it seems impracticably large; and fourth, to help avoid some of the more common mistakes encountered when computing sample sizes. PMID- 15834630 TI - Generalization of frequency discrimination learning across frequencies and ears: implications for underlying neural mechanisms in humans. AB - Frequency discrimination thresholds (FDTs) at 750, 1500, 3000, and 6000 Hz were measured in 32 normal-hearing listeners before and after each listener practiced the task for 12 h at one of the above frequencies using a single ear. Marked improvements in thresholds taking place over several hours were observed during the frequency- and ear-specific training period. Comparisons between pre- and posttraining thresholds showed large improvements at the trained frequency, but also at other frequencies. The improvements were initially slightly-but significantly-larger at the trained frequency than at untrained frequencies. However, this trained-frequency advantage disappeared rapidly during the course of the two-hour multifrequency posttraining session, suggesting rapid relearning or learning generalization across frequencies. In contrast, no significant ear specificity was found, not even at early stages of the posttraining session. These findings add to earlier results suggesting that, in humans, frequency discrimination learning is only weakly frequency-specific, and they reveal that a complete generalization across frequencies can occur rapidly with little retraining at the initially untrained frequencies. Implications regarding underlying mechanisms are discussed. PMID- 15834625 TI - Treatment of dys-immune neuropathies. AB - Several therapies are currently used in dys-immune neuropathies including steroids,plasma exchange (PE), high-dose intravenous immunoglobulins(IVIg), and immuno-suppressive agents (IS). Even if there is substantial evidence that these treatments may improve the course of the neuropathy, their effectiveness is far from being complete and is sometime hampered by the occurrence of associated side effects. In Guillain-Barre syndrome (GBS),IVIg and PE are similarly effective in accelerating the recovery but there is still little evidence that they can reduce mortality or long-term disability. Recent reports on the association of intravenous methylprednisolone or interferon-beta (IFN-beta) to IVIg did not result in significant further improvement. In chronic inflammatory demyelinating polyradiculoneuropathy(CIDP) steroids, PE, and IVIG are initially similarly effective. The short-term effect of PE and IVIgand the side effects associated with the long-term use of steroids have prompted the use of several IS, interferon and,more recently, the anti-CD20 monoclonal-antibody Rituximab, but their efficacy has still to be proved in controlled studies. The recent identification of multifocal motor neuropathy(MMN) was shortly followed by the finding of an effective therapy. Almost 80% of patients respond toIVIg whose effect needs to be maintained with periodic infusions for long periods of time, and tends to decrease after several years. Also in this condition a number of immune modulating agents have been used to reduce the frequency or improve the effectiveness of IVIg,but their efficacy has not been sofar confirmed in randomized trials. Similar conclusions can be drawn for neuropathies associated with monoclonal gammopathies where only PE and IVIg have proved to be effective in controlled studies,while the promising initial results obtained with Rituximab in neuropathy associated IgM monoclonal gammopathy awaits confirmation from controlled trials. PMID- 15834631 TI - Cardiac muscle cell cytoskeletal protein 4.1: analysis of transcripts and subcellular location--relevance to membrane integrity, microstructure, and possible role in heart failure. AB - The spectrin-based cytoskeleton assembly has emerged as a major player in heart functioning; however, cardiac protein 4.1, a key constituent, is uncharacterized. Protein 4.1 evolved to protect cell membranes against mechanical stresses and to organize membrane microstructure. 4.1 Proteins are multifunctional and, among other activities, link integral/signaling proteins on the plasma and internal membranes with the spectrin-based cytoskeleton. Four genes, EPB41, EPB41L1, EPB41L2, and EPB41L3 encode proteins 4.1R, 4.1N, 4.1G, and 4.1B, respectively. All are extensively spliced. Different isoforms are expressed according to tissue and developmental state, individual function being controlled through inclusion/exclusion of interactive domains. We have defined mouse and human cardiac 4.1 transcripts; other than 4. 1B in humans, all genes show activity. Cardiac transcripts constitutively include conserved FERM and C-terminal domains; both interact with membrane-bound signaling/transport/cell adhesion molecules. Variable splicing within and adjacent to the central spectrin/actin-binding domain enables regulation of cytoskeleton-binding activity. A novel heart specific exon occurs in human 4.1G, but not in mouse. Immunofluorescence reveals 4.1 staining within mouse cardiomyocytes; thus, both at the plasma membrane and, interdigitated with sarcomeric myosin, across myofibrils in regions close to the sarcoplasmic reticulum. These are all regions to which spectrin locates. 4.1R in human heart shows similar distribution; however, there is limited plasma membrane staining. We conclude that cardiac 4.1s are highly regulated in their ability to crosslink plasma/integral cell membranes with the spectrin-actin cytoskeleton. We speculate that over the repetitive cycles of heart muscle contraction and relaxation, 4.1s are likely to locate, support, and coordinate functioning of key membrane-bound macromolecular assemblies. PMID- 15834632 TI - Single and interacting QTLs for cholesterol gallstones revealed in an intercross between mouse strains NZB and SM. AB - Quantitative trait locus (QTL) mapping was employed to investigate the genetic determinants of cholesterol gallstone formation in a large intercross between mouse strains SM/J (resistant) and NZB/B1NJ (susceptible). Animals consumed a gallstone-promoting diet for 18 weeks. QTL analyses were performed using gallstone weight and gallstone absence/presence as phenotypes; various models were explored for genome scans. We detected seven single QTLs: three new, significant QTLs were named Lith17 [chromosome (Chr) 5, peak=60 cM, LOD=5.4], Lith18 (Chr 5, 76 cM, LOD=4.3), and Lith19 (Chr 8, 0 cM, LOD=5.3); two confirmed QTLs identified previously and were named Lith20 (Chr 9, 44 cM, LOD=2.7) and Lith21 (Chr 10, 24 cM, LOD=2.9); one new, suggestive QTL (Chr 17) remains unnamed. Upon searching for epistatic interactions that contributed to gallstone susceptibility, the final suggestive QTL on Chr 7 was determined to interact significantly with Lith18 and, therefore, was named Lith22 (65 cM). A second interaction was identified between Lith19 and a locus on Chr 11; this QTL was named Lith23 (13 cM). mRNA expression analyses and amino acid haplotype analyses likely eliminated Slc10a2 as a candidate gene for Lith19. The QTLs identified herein largely contributed to gallstone formation rather than gallstone severity. Cloning the genes underlying these murine QTLs should facilitate prediction and cloning of the orthologous human genes. PMID- 15834633 TI - Genetic analysis of anal atresia in pigs: evidence for segregation at two main loci. AB - Anal atresia is a relatively common congenital malformation that occurs in about 1 out of 5000 infants, caused by abnormal hindgut development of the embryo, often associated with other developmental anomalies (e.g., Currarino, Townes Brock, Pallister-Hall syndromes, and VATER association). Genetic analysis in human families is exceedingly difficult due to the multifactorial nature of the trait. In pigs, anal atresia occurs at a higher incidence (0.18%) than in humans. A complete genome scan (165 microsatellite markers) was performed using a backcross pedigree previously obtained by crossing affected animals from a partially inbred line, selected for a high incidence of anal atresia, with an unaffected male of a different breed (Meishan). The data set was analyzed with classical linkage (TWOPOINT) and nonparametric genetic methods (NPL, Non Parametric Linkage, and TDT, Transmission Disequilibrium Test). Both methods support association of the trait with two loci on Chromosomes 9 and 15. GLI2 (GLI Kruppel family member GLI2) was identified as a positional candidate gene based on comparative mapping; radiation hybrid mapping confirmed that this locus is located within the QTL region. PMID- 15834634 TI - Abnormal postnatal maintenance of elevated DLK1 transcript levels in callipyge sheep. AB - The underlying mechanism of the callipyge muscular hypertrophy phenotype in sheep (Ovis aries) is not presently understood. This phenotype, characterized by increased glycolytic type II muscle proportion and cell size accompanied by decreased adiposity, is not visibly detectable until approximately three to eight weeks after birth. The muscular hypertrophy results from a single nucleotide change located at the telomeric end of ovine Chromosome 18, in the region between the imprinted MATERNALLY EXPRESSED GENE 3 (MEG3) and DELTA, DROSOPHILA, HOMOLOG LIKE 1 (DLK1) genes. The callipyge phenotype is evident only when the mutation is paternally inherited by a heterozygous individual. We have examined the pre- and postnatal expression of MEG3 and DLK1 in sheep of all four possible genotypes in affected and unaffected muscles as well as in liver. Here we show that the callipyge phenotype correlates with abnormally high DLK1 expression during the postnatal period in the affected sheep and that this elevation is specific to the hypertrophy-responsive fast-twitch muscles. These results are the first to show anomalous gene expression that coincides with both the temporal and spatial distribution of the callipyge phenotype. They suggest that the effect of the callipyge mutation is to interfere with the normal postnatal downregulation of DLK1 expression. PMID- 15834635 TI - Isolation and characterization of the bovine Stearoyl-CoAdesaturase promoter and analysis of polymorphisms in the promoter region in dairy cows. AB - Conjugated linoleic acid (CLA) in milk arises through microbial biohydrogenation of dietary polyunsaturated fatty acids (PUFA) in the rumen, and by the action of mammary Stearoyl-CoA desaturase (Scd). A large variation (up to tenfold) in the concentration of this fatty acid in milk has been observed, even in cows receiving the same diet. The reasons for this variation are not well understood. In this study the bovine core promoter region was isolated by a genome walking strategy from genomic DNA Genome Walker libraries and then cloned and characterized. This core promoter sequence extended approximately 600 bp upstream of the translation start site. The presence of putative transcription factor binding sites conserved in bovine, human, and mouse promoters was observed. Evidence that this promoter fragment was functional in vivo was obtained from expression studies in a mammary cell line. The promoter sequence of the scd gene was compared between cows selected for the ability to produce high fatty acid methyl esters (FAME) (2.22-2.72) in their milk, with the same promoter region of low-FAME-producing cows (0.81-1.12). However, such comparisons of the sequences of the scd promoter region of cows producing high milk CLA compared with low CLA revealed no polymorphisms in this promoter segment. Furthermore, no sequence polymorphisms were observed among the scd promoter region of Holstein Friesian, Montbeliarde, Normande, Norwegian Red, Charlois, Limousin and Kerry breeds. PMID- 15834636 TI - Characterization of the bovine ampkgamma1 gene. AB - AMP-activated protein kinase (AMPK) represents the mammalian form of the core component of a kinase cascade that is conserved between fungi, plants, and animals. AMPK plays a major role in protecting mammalian cells from metabolic stress by switching off biosynthetic pathways that require ATP and switching on ATP-regenerating pathways. In this report, we describe the isolation and characterization of the gene for the noncatalytic bovine gamma1 subunit of AMPK. The bovine ampkgamma1 (PRKAG1) gene spans in excess of 14 kb and is located at BTA 5q21-q22. It consists of 12 exons ranging in size from 38 b to 166 b, interspersed with 11 introns that range between 97 b and 6753 b in length. The coding region of the bovine gene shares 93% and 90% nucleotide sequence similarity with its human and rat counterparts, and the bovine AMPKgamma1 protein is 98% and 95% identical to its human and rat homologs, respectively, in amino acid sequence. SNP discovery using a cattle DNA panel revealed a number of polymorphisms that may be useful for the evaluation of ampkgamma1 as a candidate gene for energy metabolism-related production traits. PMID- 15834638 TI - Exon 15 BRAF mutations are uncommon in canine oral malignant melanomas. AB - An activating mutation in codon 599 of BRAF has been identified in approximately 60% of human cutaneous nevi and melanomas, but not melanomas of mucosal origin. The purpose of this study was to determine if BRAF mutations occur in canine oral malignant melanomas. The canine BRAF gene was first cloned from normal canine testicular cDNA, and a novel previously unreported splice variant involving exon 5 was identified during this process. To screen canine melanoma samples for BRAF mutation in codon 599, cDNA and genomic DNA were isolated from canine malignant melanoma cell lines and primary tumor samples respectively, all from cases seen at the Veterinary Medical Teaching Hospital at the University of California, Davis. Polymerase chain reaction (PCR) was performed for exon 15 using primers based at the 5' end of exon 15 and the 5' end of intron 15 and the resultant products were directly sequenced. No mutations in codon 599 or exon 15 were identified in any of the 17 samples evaluated. However, all of the melanoma cell lines expressed BRAF and demonstrated high levels of basal ERK phosphorylation suggesting that dysregulation of this pathway is present. Therefore, similar to the case with human mucosal melanomas, canine oral malignant melanomas do not possess codon 599 BRAF mutations commonly identified in human cutaneous melanomas. This finding supports the notion that melanomas arising from non-sun exposed sites exhibit distinct mechanisms of molecular transformation. PMID- 15834639 TI - [Postoperative pain management]. AB - Although scientific interest in the field of pain research is unremitting and the understanding of acute pain mechanisms has eminently advanced, it is evident that clinical practice of postoperative pain management still has major deficits. Indeed, the use of regional analgesia via a catheter, e.g. epidural analgesia (EDA), or patient controlled intravenous analgesia (PCIA) has become very popular. These methods require special equipment, adequate nursing skills and professional expertise as well as the establishment of clinical procedures and an appropriate logistic setup. However, the majority of patients are not managed by EDA or PCIA and cannot be ignored due to lack of knowledge of the proper use of basic analgesic treatment. Therefore, it makes sense to establish a standardized step-concept of postoperative pain management with regular quality control integrated into a multimodal perioperative rehabilitation program. PMID- 15834640 TI - Occurrence and characterization of PEND proteins in angiosperms. AB - The PEND protein is a DNA-binding protein in the inner envelope membrane of the developing chloroplast. It consists of a short pre-sequence, an N-terminal DNA binding domain (cbZIP), a central repeat domain, and a C-terminal transmembrane domain. PEND homologs have been detected in various angiosperms, including Arabidopsis thaliana, Brassica napus, Medicago truncatula, cucumber and cherry. Monocot homologs have also been detected in barley and rice, but sequence conservation was low in monocots. PEND-related sequences have not been detected in non-flowering plants and algae. Green fluorescent protein fusions consisting of the N-terminal as well as full-length PEND homologs in A. thaliana and B. napus were targeted to chloroplasts, and localized to nucleoids and chloroplast periphery, respectively. Immunoblot analysis suggested that crucifer homologs were present in chloroplasts probably as a dimer, as in the case of pea. These results suggest that PEND protein is present in angiosperms, and the homologs in crucifers are functionally analogous to the PEND protein in pea. PMID- 15834641 TI - Preliminary report on yellow long-tailed macaques (Macaca fascicularis) at Kosumpee Forest Park, Thailand. AB - We conducted a census on long-tailed macaques (Macaca fascicularis) at Kosumpee Forest Park, Thailand, and found 15 individuals of yellow pelage color out of a total of 287 individuals. There were three troops, designated A, B and C, but the yellow macaques were found only in B and C troops. The frequency of yellow macaques was 5.2% in the total population and 4.2 and 9.9% in B and C troops, respectively. These frequencies are much higher than those in rhesus macaques (M. mulatta) reared at Cayo Santiago (0.52%). They are, however, lower than those of 10 years ago. PMID- 15834642 TI - Factors associated with drug-induced visual hallucinations in Parkinson's disease. AB - AIMS: Visual hallucinations are common in medication-treated Parkinson's disease (PD) patients. Although their etiology is unknown several factors seem to be involved in their pathogenesis. The aim of this study was to identify possible risk factors and determine clinical characteristics associated with the development of visual hallucinations in PD. METHODS: 166 consecutive patients fulfilling clinical criteria for PD were studied. During a semi-structured interview, demographic characteristics and clinical variables were recorded. Motor, cognitive and psychiatric status was also assessed. Patients with and without visual hallucinations were compared using non-parametric tests, and logistic regression was applied to significant data. RESULTS: During the month before evaluation 20.4% of our patients experienced visual hallucinations (11.4% benign, 9% malignant). Logistic regression analysis identified three factors independently associated with visual hallucinations: long duration of Parkinson's disease, dementia, and disease severity as measured by the UPDRS total score. CONCLUSIONS: Our findings indicate that apart from well established risk factors such as cognitive impairment and disease duration, disease severity is also important for the development of visual hallucinations in PD. Furthermore, the presence of bradykinesia and instability, the absence of tremor and the severity of rigidity and bradykinesia (limb and axial) may act as cofactors. PMID- 15834643 TI - Severe delayed heart failure in three multiple sclerosis patients previously treated with mitoxantrone. AB - Mitoxantrone is an approved drug for patients with worsening relapsing-remitting, secondary progressive and progressive relapsing multiple sclerosis (MS). From a cohort of 820 MS patients, 52 (6%) were treated with this drug between December 1991 and April 2003. Mitoxantrone was administered at a dose of 12 mg/m(2) once a month for three months and then at three-month intervals to reach a total cumulative dose of 144 mg/m(2). The left ventricular ejection fraction was checked by radionuclide ventriculography prior to treatment and every six months. Treatment was stopped if the ejection fraction was below 50% in two consecutive ventriculographies performed one to three months apart. Cardiotoxicity during the course of the treatment was not observed. However, three patients developed congestive heart failure 24, 39 and 80 months after the last dose of mitoxantrone. Other cardiac causes were excluded. Two of these patients had been treated previously with cyclophosphamide. All patients first recovered on medical treatment, but two worsened a few months later. One patient remained severely symptomatic in spite of optimal medical treatment. Although mitoxantrone is generally well tolerated and reduces progression of disability and clinical exacerbations, our observation of a delayed cardiotoxicity makes necessary a long term follow-up of MS patients treated with this drug. PMID- 15834644 TI - Atypical clinical presentation of intracranial hypotension: coma. PMID- 15834637 TI - Transcriptional profiling of skeletal muscle tissue from two breeds of cattle. AB - We used a 9.6 K cattle muscle/fat cDNA microarray to study gene expression differences between the longuissimus dorsi (LD) muscle of Japanese Black (JB) and Holstein (HOL) cattle. JB cattle exhibit an unusual ability to accumulate intramuscular adipose tissue with fat melting points lower than that in other breeds. The LD biopsies from three JB (Tajima strain) and three HOL animals were used in this breed comparison. Seventeen genes were identified as preferentially expressed in LD samples from JB and seven genes were found to be expressed more highly in HOL. The expression of six selected differentially expressed genes was confirmed by quantitative real-time PCR. The genes more highly expressed in JB are associated with unsaturated fatty acid synthesis, fat deposition, and the thyroid hormone pathway. These results are consistent with the increased amounts and proportions of monounsaturated fatty acids observed in the muscle of JB animals. By discovering as yet uncharacterized genes that are differentially regulated in this comparison, the work may lead us to a better understanding of the regulatory pathways involved in the development of intramuscular adipose tissue. PMID- 15834646 TI - Lambert-Eaton Myasthenic syndrome associated with sarcoidosis. PMID- 15834645 TI - Increasing normal-appearing grey and white matter magnetisation transfer ratio abnormality in early relapsing-remitting multiple sclerosis. AB - Abnormalities within normal-appearing grey and white matter (NAGM and NAWM) occur early in the clinical course of multiple sclerosis (MS) and can be detected in vivo using the magnetisation transfer ratio (MTR). To better characterize the rates of change in both tissues and to ascertain when such changes begin, we serially studied a cohort of minimally disabled, early relapsing-remitting MS patients, using NAGM and NAWM MTR histograms. Twenty-three patients with clinically definite early relapsing-remitting MS (mean disease duration at baseline 1.9 years), and 19 healthy controls were studied. A magnetisation transfer imaging sequence was acquired yearly for two years. Twenty-one patients and 10 controls completed followup. NAWM and NAGM MTR histograms were derived and mean MTR calculated. A hierarchical regression analysis, adjusting for brain parenchymal fraction,was used to assess MTR change over time. MS NAWM and NAGM MTR were significantly reduced in comparison with controls at baseline and, in patients, both measures decreased further during follow-up: (-0.10 pu/year, p=0.001 and -0.18 pu/year, p<0.001 respectively). The rate of MTR decrease was significantly greater in NAGM than NAWM (p=0.004). Under the assumption that such changes are linear, backward extrapolation of the observed rates of change suggested that NAWM abnormality began before symptom onset. We conclude that increasing MTR abnormalities in NAWM and NAGM are observed early in the course of relapsing-remitting MS. It is now important to investigate whether these measures are predictive of future disability, and consequently, whether MTR could be used as a surrogate marker in therapeutic trials. PMID- 15834647 TI - Correlation of midbrain diameter and gait disturbance in patients with idiopathic normal pressure hydrocephalus. AB - BACKGROUND AND PURPOSE: Although gait disturbance is an important feature of idiopathic normal pressure hydrocephalus (NPH), only tentative theories have been offered to explain its pathophysiology. It has been suggested that the mesencephalic locomotor region is the anatomical substrate for the development of the hypokinetic NPH gait. To investigate this possibility, we evaluated the correlation between gait disturbance and midbrain diameter. METHODS: We enrolled 21 patients with NPH and 20 age-matched control subjects for the study. The maximal diameter of the midbrain and pons, and the width of the lateral and third ventricles were measured using midsagittal T1-weighted MRI and axial T2-weighted MRI, respectively. Gait disturbance, cognitive dysfunction, and incontinence were semiquantified. RESULTS: The maximal midbrain diameter was significantly smaller in the NPH group than in the controls (14.8 +/- 0.9 vs. 17.1 +/- 0.7 mm, p < 0.001). There were inverse correlations between the midbrain diameter and the widths of the two ventricles (r = -0.562, p = 0.008 for the third ventricle, and r = -0.510, p = 0.018 for the lateral ventricle). The severity of gait disturbance was negatively correlated with the midbrain diameter (r = -0.598, p = 0.004), but the degree of cognitive dysfunction and incontinence showed no significant correlation with midbrain diameter or ventricular width. CONCLUSIONS: This study suggests that midbrain atrophy is significantly associated with gait disturbance in NPH. PMID- 15834648 TI - Mills syndrome with dementia: broadening the phenotype of FTD/MND. PMID- 15834649 TI - Glial cell cytoplasmic inclusions in SCA2 do not express alpha-synuclein. PMID- 15834650 TI - Release of brain-type and heart-type fatty acid-binding proteins in serum after acute ischaemic stroke. AB - This study aimed at an analysis of the release of Braintype and Heart-type Fatty Acid- Binding Proteins (B-FABP and HFABP) in acute ischaemic stroke and their potential value as neurobiochemical markers of brain damage. We investigated 42 consecutive patients admitted within 6 hours after ischaemic stroke. Serial venous blood samples were taken hourly between 1 to 6 hours, and at 12, 18, 24, 48, 72, 96, and 120 hours after stroke onset. In all patients lesion topography was assessed and infarct volume was calculated. The neurological deficit was quantified by the National Institutes of Health stroke scale score, and functional outcome was assessed with the modified Rankin Scale 3 months after stroke. H-FABP and B-FABP concentrations showed peak values already 2 to 3 hours after stroke onset and remained elevated up to last measurements at 120 hours.Unlike BFABP, early H-FABP concentrations were significantly associated with the severity of the neurological deficit and the functional outcome. High H FABP release was associated with large infarction on CT. Our study shows for the first time quantitative data of serum BFABP and H-FABP being elevated early in acute ischaemic stroke indicating that especially H-FABP might have the potential to be a rapid marker of brain damage and clinical severity. As both FABPs indicate damage to neuronal and glial tissue but are not specific for cerebral infarction, further investigations are needed to better understand the prolonged release of both in ischaemic stroke which is in contrast to the transient increase after myocardial infarction and can not be explained by their renal extraction. PMID- 15834651 TI - X-linked paroxysmal dyskinesia and severe global retardation caused by defective MCT8 gene. AB - We previously reported two unrelated boys aged 3 and 8 years with mutations in the thyroid hormone transporter gene MCT8 resulting in severe global retardation and an uncommon pattern of thyroid hormone abnormalities. We now further describe an unusual neurological phenotype associated with these mutations, namely paroxysmal kinesigenic dyskinesias (PKD), provoked by certain stimuli including changing of their clothes or diapers. It is not clear how the MCT8 defect causes PKDs. PKDs have been previously noted in patients with thyroid abnormalities. This novel X-linked condition widens the spectrum of secondary PKDs. PMID- 15834652 TI - Local tissue anisotropy decreases in cerebellopetal fibers and pyramidal tract in multiple system atrophy. AB - BACKGROUND: One of the cardinal features in multiple system atrophy (MSA) is the white matter pathology: loss of myelin, astrocytosis, and glial cytoplasmic inclusions. The pathological changes of tissue microstructure can modify the diffusion behavior of water molecules, which can be assessed by diffusion tensor imaging (DTI). OBJECTIVES: To explore the hypothesis of white matter degeneration in MSA. METHODS: We studied 11 patients with clinically probable MSA and 10 age matched controls. DTI was performed in both groups to measure fractional anisotropy (FA) in various regions of interest: the inferior cerebellar peduncle (ICP), middle cerebellar peduncle (MCP), superior cerebellar peduncle (SCP), basis pontis, internal capsule, and corpus callosum. RESULTS: FA values in SCP and corpus callosum showed no significant difference between the MSA group and controls. By contrast, FA values decreased in the MSA group in the MCP, basis pontis and internal capsule. In addition, FA values in the MCP were negatively correlated with ataxia severity in the MSA group. CONCLUSION: The areas showing decreased tissue anisotropy in DTI corresponded well with pathologically vulnerable areas in MSA. In addition, the local tissue anisotropy of MCP decreased in accordance with functional disability. These observations implied that DTI is a feasible method for in vivo evaluation of the white matter pathology in MSA. PMID- 15834653 TI - Characterization of Helicoverpa armigera nucleopolyhedrovirus orf33 that encodes a novel budded virion derived protein, BV-e31. AB - Homologues of Helicoverpa armigera nucleopolyhedrovirus (HearSNPV) orf33 are found in all 22 completely sequenced members of the lepidopteran nucleopolyhedroviruses and granuloviruses, but so far their functions are unknown. In this report, we describe the characterization of HearSNPV orf33 (ha33). Northern blot analysis showed a single transcript of ha33 of approximately 0.7 kb was transcribed beginning at 3 h post-infection in infected Helicoverpa zea cells (HzAM1) and the gene product could be detected as early as 6 h post-infection by western blot analysis using a rabbit derived polyclonal antibody, suggesting it was an early gene. Western blot analysis also demonstrated the ha33 protein in infected cells was a 31 kDa protein, larger than the theoretical size of 28.4 kDa, and located in the envelope fraction of budded virions (BVs). The results suggested that HearSNPV ha33 gene is a functional gene that encodes a novel structural protein of baculovirus BVs, BV-e31. PMID- 15834654 TI - The inhibitor of apoptosis gene (iap-3) of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV) encodes a functional IAP. AB - Programmed cell death or apoptosis is one of the defense mechanisms used by insect cells in response to baculovirus infection. Baculoviruses harbour antiapoptotic genes to prevent apoptosis and to maintain the normal course of infection. In this work, we showed that, like other baculoviruses, Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV) has a functional inhibitor of apoptosis gene (iap-3). The iap-3 gene was cloned, sequenced and its transcription confirmed by RT-PCR. The putative iap-3 gene of the baculovirus AgMNPV has 864 nucleotides and codes an ORF of 287 amino acids. We have found two BIR motifs (baculoviral iap repeats) at the amino-terminal region and a carboxi terminal RING finger motif. The IAP-3 protein of AgMNPV is closely related to IAP 3 proteins of baculoviruses and lepidopteran IAPs, with most amino acid identity (75%) with the IAP-3 protein of CfDefNPV (Choristoneura fumiferana DEF nucleopolyhedrovirus). Transcriptional analysis of the AgMNPV iap-3 gene showed that iap-3-specific transcripts could be detected early and late in the infection. The iap-3 gene of AgMNPV was shown to encode a functional IAP since insect cells transfected with increasing amounts of a plasmid containing the iap 3 of AgMNPV showed increased resistance to apoptosis induced by a AgMNPV mutant virus. PMID- 15834655 TI - The requirements and mechanism for capsid assembly and budding of bovine foamy virus. AB - Little is known about assembly of non-primate foamy virus (FV) such as bovine foamy virus (BFV). To help determine the requirements for assembly of BFV, we constructed BFV-Gag expression plasmids containing all or part of the gag gene, with or without modification by addition of myristate (Myr). Each construct was transfected alone, and with pFenv, into Sf-9 insect cells. The results showed that only the entire Gag could transit through nucleus, which is required for BFV viral assembly in the cytoplasm. Unlike other retroviruses (but like primate foamy viruses), BFV requires the coexpression of the Env protein for viral particle budding. In the case of BFV, this occurs at the plasma membrane rather than the endoplasmic reticulum (ER), due to lack of a functional ER retrieval signal (ERRS). The results also showed that addition of a Myr-membrane targeting signal to the C-terminus of Gag could restore the budding from plasma membrane, implying that Myr-membrane targeting signal could substitute for Env protein in budding. PMID- 15834656 TI - Heterogeneity in codon usages of sobemovirus genes. AB - When conventional phylogenetic trees were built using 14 genome sequences of 9 sobemoviruses, two main lineages were apparent: monocot-infecting viruses and dicot-infecting viruses. To investigate whether members of the genus Sobemovirus originated from monocot hosts or from dicot hosts, we constructed relationship trees based on Relative Synonymous Codon Usage (RSCU) of the viruses. The RSCU relationship trees grouped the monocot-infecting and dicot-infecting viruses even better than the genome phylogenetic trees. The RSCU approach also enabled direct comparisons among viral and host species. When host species were added into the RSCU tree, the viral species clustered with the monocot hosts, indicating codon usage homologies to monocots. The stability of the RSCU tree was improved when RSCU values were calculated for individual viral open reading frames (ORFs). Most interestingly, the codon usages of the viral ORF-2 that encodes the replicase showed affinity to that of the plants whereas codon usages of the other viral ORFs were not relevant to the host species. All ORF-2s from 3 monocot viruses and 4 out of 6 dicot viruses had greater RSCU affinities to sequences of ORFs in monocot than to dicot hosts, possibly indicating that ORF-2, and therefore the replicase module of sobemovirus has a monocot origin. PMID- 15834659 TI - The stationary phase model of aging in yeast for the study of oxidative stress and age-related neurodegeneration. AB - Understanding the biochemical and genetic alterations that occur during the aging of post-mitotic cells is critical for understanding the etiology of abnormalities observed during the aging of the central nervous system (CNS). While many theories for cellular aging exist, the free radical theory of aging has proved useful in explaining multiple aspects of post-mitotic cell aging, including the aging of neuronal cells. It is well established that Saccharomyces cerevisiae are an invaluable model system for exploring the regulation of aging in actively dividing cells, but increasing evidence suggests that the chronological lifespan or stationary phase model of aging in S. cerevisiae may also be useful for understanding the aging process in post-mitotic cells. Interestingly, the stationary phase model of aging in S. cerevisiae recapitulates many pathological alterations observed during neuronal aging, including evidence for increased oxidative stress and proteasome inhibition. Studies using proteins relevant to multiple neurodegenerative conditions (prion, alpha-synuclein, huntingtin) have demonstrated the utility of S. cerevisiae as a model system for understanding the genetic regulation of protein aggregation and cell death. Taken together, these data highlight the potential importance of using S. cerevisiae as a model system with which to explore the molecular basis for neuronal alterations observed in normal brain aging as well as multiple age-related diseases of the CNS. PMID- 15834660 TI - Effect of insulin and growth hormone on rat heart and liver oxidative stress in control and caloric restricted animals. AB - In order to know if insulin-like signalling is involved in the control of oxidative stress in mammalian tissues in relation to aging, ad libitum-fed and caloric restricted Wistar rats were treated during 2 weeks with GH and insulin. The most consistent effect of the hormonal treatments was an increase in plasma IGF-1 levels. Caloric restriction during 6 weeks decreased ROS generation and oxidative DNA damage in heart mitochondria and this was reversed by insulin treatment. The decrease in oxidative damage to liver nuclear DNA induced by caloric restriction was also reversed by GH and insulin. In the liver, however, insulin and GH decreased mitochondrial ROS generation while they increased oxidative damage to mitochondrial DNA. GH and insulin decreased three different markers of oxidative modification of liver proteins, while they increased lipoxidation-dependent markers. This last result is related to the increase in phospholipid unsaturation induced in the liver by both hormones. The results suggest that the idea that insulin-like signalling controls oxidative stress in mammals cannot be generalized since both prooxidant and protective effects of GH and insulin are observed depending on the particular parameter and tissue selected. PMID- 15834661 TI - The effect of age and calorie restriction on HIF-1-responsive genes in aged liver. AB - Hypoxia inducible factor-1 (HIF-1) regulates transactivation of several genes in response to hypoxia condition. We explore hepatic HIF-1 responsive gene regulation during aging and the age-related changes of the HIF-1 related gene activation in young and old rats. Results indicate that the aging process induces the activation of HIF-1alpha, which is accompanied by increased HIF-1 DNA binding. This increased binding activity is accompanied by the increase of HIF-1 dependent genes, heme oxygenase-1 (HO-1), vascular endothelial growth factor (VEGF), erythropoietin (EPO), and inducible nitric oxide synthase (iNOS), which all showed remarkable up-regulation during aging process. In contrast, the increased HIF-1 related gene expression was effectively blunted by the anti oxidative action of calorie restriction in aged rat liver. We propose that age related HIF-1 binding activity may well be influenced by the increased pro oxidative conditions of aged animals, which up-regulate HIF-1-dependent gene expression. PMID- 15834662 TI - Testing the "garbage" accumulation theory of ageing: mitotic activity protects cells from death induced by inhibition of autophagy. AB - Imperfect autophagic degradation of oxidatively damaged macromolecules and organelles (so-called biological "garbage") is considered an important contributor to ageing and consequent death of postmitotic (non-dividing) cells, such as neurons and cardiac myocytes. In contrast, proliferating cells apparently escape senescence by a continuous dilution and repair of damaged structures during division. Postmitotic ageing can be mimicked and studied in cultures of potentially dividing cells if their mitotic activity is inhibited. To test the "garbage accumulation" theory of ageing, we compared survival of density dependent growth-arrested (confluent) and proliferating human fibroblasts and astrocytes following inhibition of autophagic sequestration with 3-methyladenine (3MA). Exposure of confluent fibroblast cultures to 3MA for two weeks resulted in a significantly increased proportion of dying cells compared to both untreated confluent cultures and dividing cells with 3MA-inhibited autophagy. Similar results were obtained when autophagic degradation was suppressed by the protease inhibitor leupeptin. In 3MA- or leupeptin-exposed cultures, dying cells were overloaded with undegraded autofluorescent material. The results support a key role of biological lysosomal "garbage" accumulation in the triggering of ageing and death of postmitotic cells, as well as the anti-ageing role of cell division. PMID- 15834663 TI - Effects of ovariectomy and growth hormone administration on body composition and vascular function and structure in old female rats. AB - Aging and estrogen-deprivation induce deleterious effects on body composition and vascular function in females. On the other hand, growth hormone (GH), whose production is reduced by age, exerts several vascular effects. The aim of this study was to investigate the effect of long-term estrogen deprivation and GH administration on body composition, vascular function and structure in aged female rats. METHODS: Twelve female Wistar rats were ovariectomized at 10 months of age. At 20 months of age, half of the ovariectomized rats were treated with GH for 4 weeks. The remaining ovariectomized rats animals and one group of six intact females were used as control groups. After the treatment period, animals were sacrificed and Specific Gravity Index (SGI) and periuterine fat weigh, as well as vascular reactivity and morphometry in aortic rings, were studied. RESULTS: No significant differences were found in SGI and periuterine fat weigh between ovariectomized and intact control rats. SGI was significantly increased by GH, and periuterine fat was reduced by the treatment. Dose-dependent relaxing responses to acetylcholine and isoproterenol were significantly diminished in ovariectomized rats as compared with intact animals, and GH treatment improved these responses. Ovariectomized animals showed significantly higher contracting responses to phenylephrine, acetylcholine + L-NAME and angiotensin-I than intact rats, and treatment with GH reduced them significantly. Media cross-sectional area was increased in ovariectomized rats as compared to intact animals, and GH reduced this area, but differences did not reach significance. CONCLUSION: GH has beneficial effects in body composition and endothelial function in old ovariectomized female rats. PMID- 15834664 TI - Maternal and grandmaternal age effects on developmental instability and wing size in parthenogenetic Drosophila mercatorum. AB - Age is known to have a large effect upon fertility and survival. Here we study the impact of maternal and grandmaternal age on wing size and developmental instability (DI) in offspring using a parthenogenetic strain of Drosophila mercatorum. This enabled us to cancel out any genetic variance between individuals. The mean centroid size of the wings was estimated in offspring from four maternal/grandmaternal age groups. Further, DI was calculated by measuring phenotypic variability and correcting this index for the bias produced by the environmental variability. The offspring developed from eggs laid by mothers belonging to four different age groups with young (Y) or old (O) mothers and grandmothers. The age groups are: YY, YO, OY and OO, with the first letter referring to the age of the grandmother and the second letter to the age of the mother. We consider flies between three to six days of age as young and flies between 15 and 18 days of age as old. We found that environmental variability was present in this study and therefore the correction for its bias on phenotypic variability was shown to be necessary for a correct interpretation of the results. DI displayed, for most of the traits investigated, a significantly higher level in the OY and OO age groups, as compared to the YY and YO age groups. The mean centroid size of the wings were in almost all cases significantly different among the age groups with the highest mean value in the OY group for all traits. This investigation shows that offspring are affected by maternal age and that this effect can be transmitted maternally across more than one generation. PMID- 15834665 TI - Exercise and hormesis: oxidative stress-related adaptation for successful aging. AB - The hormesis theory purports that biological systems respond with a bell-shaped curve to exposure to chemicals, toxins, and radiation. Here we extend the hormesis theory to include reactive oxygen species (ROS). We further suggest that the beneficial effects of regular exercise are partly based on the ROS generating capability of exercise, which is in the stimulation range of ROS production. Therefore, we suggest that exercise-induced ROS production plays a role in the induction of antioxidants, DNA repair and protein degrading enzymes, resulting in decreases in the incidence of oxidative stress-related diseases and retardation of the aging process. PMID- 15834666 TI - Biophysics of aging and therapeutic interventions by entropy-variation systems. AB - The cell is thermodynamically an open system and aging is characterized by an increasingly higher structural disorder (increase of entropy) and functional loss. If a variation of negative entropy is introduced by an external source, an anti-clockwise effect leading to regenerative processes and/or increase of the functional reserve supporting regenerative tissue changes is theoretically expected. The achievement of a negative variation of entropy is the main principle of a new technology which implies an exogenous delivery of energy with higher performance than the physiological production. Promising clinical experiences in liver cirrhosis and in long-standing scarring lesions seem to confirm the clinical applicability of the theoretical model. PMID- 15834667 TI - Informed consent of very old patients and modern genomics. PMID- 15834674 TI - Paraperitoneal indirect inguinal bladder hernia: MR demonstration. AB - Inguinal or inguinoscrotal herniation of the bladder is not uncommon and has been estimated to comprise 1% to 3% of all inguinal hernias. The appearance of hernias on ultrasonography, intravenous pyelography, cystography, and computed tomography has been described previously but no instance of correlation with magnetic resonance (MR) imaging has been documented. Nevertheless, herniated bladders can be encountered fortuitously during abdominal MR imaging, and the radiologist should be familiar with their appearance. We present a case of unsuspected paraperitoneal indirect inguinal bladder herniation demonstrated by MR. Appearance on MR is characteristic, and this modality may be useful for differentiating the several types of inguinal hernias of the bladder because of its superior soft tissue contrast. In addition, MR imaging can be used to perform imaging in any plane and dynamic examinations during straining. PMID- 15834675 TI - Ileosigmoid knot: case report and CT findings. AB - Ileosigmoid knot (ISK) is an unusual clinical entity of small bowel obstruction in which the ileum wraps around the base of the sigmoid colon and forms a pseudo knot. We present the case of a 75-year-old male in whom ISK was the definitive computed tomographic finding. ISK should be considered in differential diagnosis of patients who present with ileus. PMID- 15834676 TI - Ruptured gastrointestinal stromal tumors: radiologic findings in six cases. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are characterized by the expression of c-KIT (antigen CD 117) and are the most common mesenchymal tumors of the digestive tract. An important complication, although infrequently described in the literature, is the rupture of these tumors with accompanying hemoperitoneum. METHODS: We performed a retrospective evaluation of the clinical history and radiologic records of 23 patients with a diagnosis of GIST and anatomopathologic and immunohistochemical confirmation at our hospital between 1999 and 2004. RESULTS: In five cases there was rupture of the primary tumor (four gastric and one jejunal). In all five cases ultrasonographic and computed tomographic examinations showed a heterogenic tumor of laminated or whirled appearance, associated with echogenic or dense ascites. No relation was found between histologic criteria of malignancy and the rupture. Four patients underwent surgical intervention, three of them urgently. Two of five patients died. There was a sixth case with rupture of a hepatic metastasis, with accompanying hemoperitoneum and subcapsular hematoma. This patient died at 3 months, after recurrence of bleeding. CONCLUSIONS: The finding of a heterogeneous tumor of laminated or whirled appearance associated with ascites with characteristics compatible with hemoperitoneum in an appropriate context must lead to a suspicion of the existence of a ruptured GIST. PMID- 15834677 TI - Female genitalia hydatidosis herniating to the inguinal canal. AB - Hydatid disease of the female genitalia presents a rare problem in gynecology formerly limited to only a few parts of the world. Primary pelvic hydatidosis is uncommon, and its pathogenesis remains unclear. We report one case of primary hydatid cysts and another case of secondary hydatid cysts of the female genitalia that herniated to the inguinal canal. PMID- 15834678 TI - Endoleak and endotension following open abdominal aortic aneurysm repair: a report of two cases. AB - Anastomotic leaks and perigraft seroma are uncommon complications following open repair of abdominal aortic aneurysms; in particular, it is rare for them to cause recurrence and expansion of the aneurysm sac. We report a case of anastomotic leak presenting as type 1 endoleak and another of perigraft seroma causing recurrent pressurization, endotension, and expansion of the original aneurysm sac. PMID- 15834679 TI - Compartment syndrome after compression sclerotherapy. AB - Acute limb compartment syndrome is a condition in which raised pressure within a closed fascial space reduces capillary perfusion below a level necessary for tissue viability. Although it is a rare but potentially disastrous complication of orthopedic injury to the extremities, it may occur spontaneously without a history of trauma, and any insult that tends to increase resistance to flow in the capillary bed in any anatomical situation may result in a compartment syndrome. We report an extremely rare case of compartment syndrome following compression sclerotherapy. PMID- 15834680 TI - Selective delayed management of blunt traumatic left common carotid artery injury using hypothermic circulatory arrest. AB - Most traumatic carotid artery aneurysms occur at or close to its bifurcation, and traumatic aneurysm of the intrathoracic carotid arteries are rare. We describe a case of false aneurysm at the origin of the left common carotid artery (LCCA) after blunt trauma. A 53-year-old man suffered a blow from a broken steel plate, which flew from a working concrete crusher over his neck when he looked down the machine. Chest computed tomography revealed aneurysm of the LCCA, and aortic arch arteriography demonstrated a false aneurysm of about 3 x 5 cm at the origin of the LCCA, with loss of arterial continuity and abnormal tortuosity above the aneurysm. An ascending aorta to LCCA bypass graft was placed during the cooling period of cardiopulmonary bypass, and mattress sutures were placed in the normal aorta to close the origin of the LCCA under hypothermic circulatory arrest because of the extreme danger of dissection. The LCCA was transected partially at its origin from the aorta. We speculated that the direct lifting force which caused the carotid artery to move upward might produce a tear at the junction of the LCCA and the aortic arch. PMID- 15834681 TI - Revascularization using arterial conduits for abdominal angina due to isolated and spontaneous dissection of the superior mesenteric artery. AB - Isolated spontaneous dissection of the superior mesenteric artery is rare, and the surgical treatment is not established. We performed successfully total revascularization using arterial conduits (left radial artery and right epigastric artery) and present here the first report to our knowledge of a surgically treated case. PMID- 15834682 TI - Acute supramesenteric thrombosis of an abdominal aortic aneurysm with deleterious embolism: a case report. AB - A 55-year-old man was admitted with severe pain, paralysis of both legs and absent femoral pulses. Computed tomographic scan demonstrated a 6 cm juxtarenal abdominal aortic aneurysm (AAA) with thrombosis starting at the level of the celiac trunk. At immediate operation, thrombectomy of visceral arteries was performed and distal neovascularization was achieved with a bifurcated prosthesis. It was revealed that all major arteries were occluded with debris. Embolectomy did restore flow in major vessels, but organ perfusion was not achieved due to occlusion of smaller vessels. The patient died with multiorgan failure. This is the first description in the literature of an acutely thrombosed AAA at the supramesenteric level. PMID- 15834683 TI - Ex vivo repair and renal autotransplantation for complex renal artery aneurysms in a solitary kidney. AB - Renal artery aneurysm (RAA) is a rare clinical entity with an incidence of 0.015 1%. Indications for interventional or surgical repair of RAAs are expanding aneurysms, diameter >2.5 cm, intractable renovascular hypertension, dissecting RAA, hematuria, and renal infarction after distal embolization. Interventional insertion of a stent graft as well as aortorenal bypass implantation are both low risk procedures in simple aneurysms of the proximal renal artery. However, complex distal renal aneurysms involving several renal artery branches require not only an excellent result of vascular reconstruction, but also a surgical technique offering maximal protection for the kidney during the ischemic period. Here, we present a case of a solitary kidney with two consecutive RAAs of segmental renal artery branches (type 2 RAA). A surgical strategy including renal explantation, ex vivo renal preservation, ex vivo reconstruction of the renal artery, and renal heterotopic autotransplantation was successfully applied. The technique of ex vivo repair is a safe and effective surgical procedure in this clinical setting. PMID- 15834684 TI - Paclitaxel impairs endothelial cell adhesion but not cytokine-induced cellular adhesion molecule expression. AB - Local delivery of antiproliferative agents using drug-eluting stents has become a productive area of research for preventing in-stent restenosis. Recently, the microtubule stabilizing drug paclitaxel has been used to coat stents. While the actions of paclitaxel on smooth muscle are well documented, effects on endothelial cells (ECs) are largely unknown. Nevertheless, restoration of EC function is a critical step in repairing the vascular lesion. We assessed the effects of paclitaxel by examining three events that are critical in controlling the severity of vascular injury: (1) adhesion of ECs to matrix proteins, (2) EC migration, and (3) cytokine-stimulated cellular adhesion molecule (CAM) expression on the surface of ECs. Paclitaxel inhibited both EC adhesion and migration of ECs; however, it had no effect on tumor necrosis-stimulated CAM expression on ECs. The mechanisms of paclitaxel action on matrix adhesion and migration are not clear, but protein kinase C and myosin light chain kinase do not appear to play a role as they are unaffected by treatment of the cells with paclitaxel. On the other hand, the MAP kinase ERK1/2 is modestly inhibited by paclitaxel. While paclitaxel-coated endovascular stents may prevent smooth muscle proliferation, their attenuation of EC migration and adhesion to the lesion coupled with an inability to reduce cytokine-induced CAM expression on ECs may limit their effectiveness. PMID- 15834685 TI - Electrogenic glutamate transporters in the CNS: molecular mechanism, pre-steady state kinetics, and their impact on synaptic signaling. AB - Glutamate is the major excitatory neurotransmitter in the mammalian CNS. The spatiotemporal profile of the glutamate concentration in the synapse is critical for excitatory synaptic signalling. The control of this spatiotemporal concentration profile requires the presence of large numbers of synaptically localized glutamate transporters that remove pre-synaptically released glutamate by uptake into neurons and adjacent glia cells. These glutamate transporters are electrogenic and utilize energy stored in the transmembrane potential and the Na+/K+-ion concentration gradients to accumulate glutamate in the cell. This review focuses on the kinetic and electrogenic properties of glutamate transporters, as well as on the molecular mechanism of transport. Recent results are discussed that demonstrate the multistep nature of the transporter reaction cycle. Results from pre-steady-state kinetic experiments suggest that at least four of the individual transporter reaction steps are electrogenic, including reactions associated with the glutamate-dependent transporter halfcycle. Furthermore, the kinetic similarities and differences between some of the glutamate transporter subtypes and splice variants are discussed. A molecular mechanism of glutamate transport is presented that accounts for most of the available kinetic data. Finally, we discuss how synaptic glutamate transporters impact on glutamate receptor activity and how transporters may shape excitatory synaptic transmission. PMID- 15834686 TI - Association of alpha-dystrobrevin with reorganizing tight junctions. AB - Alpha-dystrobrevin (alpha-DB) has been described primarily as a cytoplasmic component of the dystrophin-glycoprotein complex in skeletal muscle cells. Isoforms of alpha-DB show different localization in cells and tissues; at basolateral membranes in epithelial cells, dystrobrevins mediate contact with the extracellular matrix, peripheral and transmembrane proteins and the filamentous actin cytoskeleton. Beside their structural role, alpha-DBs are assumed to be important in cell signalling and cell differentiation. We have primarily assessed the role of alpha-DB in two epithelial cell lines (MDCK I, HT 29), which represent different developmental stages and exhibit distinct permeability characteristics. Using a polyclonal anti-alpha-DB antibody, we have investigated its expression, localization and association with tight junction (TJ)- associated proteins (ZO-1, occludin) before and after protein kinase C (PKC) activation with phorbol myristate acetate. Distinct subsets of alpha-DB isoforms were detected in the two cell lines by immunoblotting. In both cell lines there was submembranous localization of alpha-DB both apically and basolaterally, shown with confocal imaging. PKC activation caused a reorganization of TJ, which was parallel to increased localization of alpha-DB to TJ areas, most pronounced in MDCK I cells. Moreover, actin and ZO-1 co-immunoprecipitated with a-DB, as displayed with immunoblotting. Our findings suggest that a-dystrobrevin specifically is associated with the tight junctions during their reorganization. PMID- 15834687 TI - New concept of spare receptors and effectors. AB - The present study provides a new concept of the spare receptor. Model [A]: 1) Several receptors connect with an effector; 2) if an agonist occupies one of the receptors connecting with one effector, the effector fully functions. When the number of receptors connecting with one effector is "m", the relationship between the functional effectors (E) and the concentration of agonists ([a]) is as follows: [formula: see text] where Rt is the total number of receptors and Kd is the agonist dissociation constant from the receptor. Model [B]: 1) Several receptors connect with an effector; 2) only when agonists occupy all of the receptors connecting with one effector, the effector functions. The relationship between E and [a] is as follows: [formula: see text] If m=1, equations (I) and (II) are exactly the same as the Michaelis-Menten equation. If m is larger than 1, the apparent saturation in the effector efficiency becomes larger in Model [A], and smaller in Model [B], respectively. The dissociation of the fractional efficiency of effectors from the fractional binding of agonists to receptors becomes larger as m becomes larger in both models. Further, we propose a variable model, including the concept of agonist-occupancy-dependent stability in the functional conformation change of the effector; only when more than j pieces of receptors connecting with one effector are occupied by agonists, the effector functions (Model [M]). The relationship between E and [a] is as follows: [formula: see text] PMID- 15834688 TI - A mathematical model of electrolyte and fluid transport across corneal endothelium. AB - To predict the behavior of a transporting epithelium by intuitive means can be complex and frustrating. As the number of parameters to be considered increases beyond a few, the task can be termed impossible. The alternative is to model epithelial behavior by mathematical means. For that to be feasible, it has been presumed that a large amount of experimental information is required, so as to be able to use known values for the majority of kinetic parameters. However, in the present case, we are modeling corneal endothelial behavior beginning with experimental values for only five of eleven parameters. The remaining parameter values are calculated assuming cellular steady state and using algebraic software. With that as base, as in preceding treatments but with a distribution of channels/transporters suited to the endothelium, temporal cell and tissue behavior are computed by a program written in Basic that monitors changes in chemical and electrical driving forces across cell membranes and the paracellular pathway. We find that the program reproduces quite well the behaviors experimentally observed for the translayer electrical potential difference and rate of fluid transport, (a) in the steady state, (b) after perturbations by changes in ambient conditions HCO3-, Na+, and Cl- concentrations), and (c) after challenge by inhibitors (ouabain, DIDS, Na+- and Cl(-)-channel inhibitors). In addition, we have used the program to compare predictions of translayer fluid transport by two competing theories, electro-osmosis and local osmosis. Only predictions using electro-osmosis fit all the experimental data. PMID- 15834689 TI - [New guidelines for treatment of hypertension]. AB - Treatment of high blood pressure is a central element in prevention of cardiovascular diseases. The new classification of hypertension takes into consideration the close association between blood pressure level and cardiovascular risk and designates blood pressure between 140/90 and 130/80 mmHg as high-normal so that blood pressure <140/90 mmHg should always be the goal. The targeted blood pressure levels are also defined by the extent of end-organ damage already present. The therapeutic objective in patients with diabetes mellitus is a blood pressure level of 130/80 mmHg and in patients with kidney disease and proteinuria 125/75 mmHg. The five substance groups of diuretics, beta-blockers, calcium antagonists, ACE inhibitors, and angiotensin receptor blockers are recommended for primary treatment. In addition to the antihypertensive properties, substance-specific effects of ACE inhibitors and angiotensin receptor blockers have been described. Primarily, instead of monotherapy low-dose combination therapy can also be judicious. PMID- 15834690 TI - Hormetic protection of Drosophila melanogaster middle-aged male flies from heat stress by mildly stressing them at young age. AB - Previous studies have shown that exposing flies to hypergravity (3g or 5g) for the first 2 weeks of adult life slightly increases longevity of male flies and survival time at 37 degrees C for both sexes, and delays an age-linked behavioral change. The present experiment tested whether the hypergravity could also protect flies from four successive deleterious non-lethal heat shocks at 4 and 5 weeks of age. Males that lived in hypergravity for the first 2 weeks of adult life lived slightly longer (ca. +15% or 1.2 day) after heat shocks (30 min or 45 min at 37 degrees C) than flies that always lived at 1g, but this positive effect of hypergravity was not observed in females. Therefore, hypergravity exposure at young age can help the male flies recovering from a heat shock at older ages. PMID- 15834691 TI - A large Cretaceous theropod from Patagonia, Argentina, and the evolution of carcharodontosaurids. AB - The Cretaceous Carcharodontosauridae is the latest clade of carnosaurs, including the largest predatory dinosaurs yet recorded. Albeit spectacular for their size, the skeletal anatomy of these theropods remains poorly-known, and their diversity was until recently restricted to two Cenomanian species: the highly derived Giganotosaurus carolinii, from southern South America, and the incompletely known Carcharodontosaurus saharicus, from northern Africa. Here we describe an older and basal member of the group, Tyrannotitan chubutensis gen. et sp. nov., from Aptian strata of Patagonia, Argentina. The new taxon gives new insights into the systematics and evolution of carcharodontosaurids and offers a better understanding of the evolution of Southern theropod faunas. We suggest that carcharodontosaurids radiated in Gondwana sharing with spinosaurids the role of top-predators until their extinction in Cenomanian-Turonian times. During this interval, the diplodocoid sauropods and giant titanosaurians went extinct (probably as part of a global-scale crisis), and the smaller abelisaurid theropods took dominance, reigning until the end of the Cretaceous. Electronic Supplementary Material is available. PMID- 15834692 TI - A homolog of the hydrolase Notum is expressed during segmentation and appendage formation in the Central American hunting spider Cupiennius salei. AB - The hydrolase Notum (also known as Wingful) has been implicated in shaping the distribution gradient of the morphogen Wingless in Drosophila by modifying the Wingless-binding heparan sulfate proteoglycan (HSPG) core glypicans Dally and Dally-like. Here we report on the expression of a Notum homolog during the embryonic development of the spider Cupiennius salei. Notum is expressed in two to three stripes in the posterior region of the germband where new segments are formed. At this location no Wingless expression is present, suggesting that Notum may be involved in the regulation of another HSPG-binding morphogen, possibly Hedgehog. In older segments, however, expression of Notum and Wingless roughly coincides. In the appendages, Notum is expressed in ventral mesodermal cells, directly adjacent to the Wingless expressing ectodermal cells. This could indicate a role for the mesoderm in regulating morphogen gradient formation in the ectoderm. PMID- 15834693 TI - [Tumour necrosis factor receptor-associated periodic syndrome. A rare differential diagnosis of multiple sclerosis]. AB - Tumour necrosis factor receptor-associated periodic syndrome (TRAPS), a rare autosomal dominant disorder, is characterised by recurrent attacks of fever, myalgias, and abdominal pain. However, manifestations in the central nervous system are hardly known. We describe a family in which one of three affected members developed central nervous system symptoms. First diagnosed as multiple sclerosis (MS), the demyelinisation seems to be a feature of TRAPS rather than MS. The syndrome is discussed as a rare differential diagnosis of MS. PMID- 15834694 TI - [Ultrasound contrast agents and prostate cancer]. AB - Prostatic carcinoma is the most frequent malignant disease in men and associated with very high mortality. The diagnostic work-up of prostatic carcinoma is based on tests to determine the level of prostate-specific antigen (PSA), digital rectal examination, and transrectal sonography. Due to diagnostic limitations, ultrasound-guided prostate biopsy is the method of choice for diagnosis of prostatic carcinoma. New imaging technologies allow detection of prostatic carcinoma, thus facilitating removal of specific biopsy specimens from these regions. Introduction of ultrasound contrast agents ("echo signal enhancers") significantly increased the diagnostic potential of this method, making it possible to visualize tumor vascularization. PMID- 15834695 TI - [Concealment of benefits by a physician working in his/her own practice is punishable by law. Verdict of the Higher Regional Court, Hamm, 22 December 2004]. PMID- 15834696 TI - Molecular characterization of new waxy mutants identified in bread and durum wheat. AB - Recently, electrophoretic analyses of waxy proteins in several hexaploid and tetraploid wheat accessions from worldwide collections have permitted the identification of new variants at the waxy loci, including allelic forms with different mobilities and partial null types. In this paper, the molecular characterization of mutated waxy loci in four bread wheat cultivars (two lacking the Wx-B1 and two lacking the Wx-D1 protein, respectively) and in four durum wheat cultivars (one lacking Wx-A1 and the remainder with Wx-B1 proteins showing different electrophoretic mobilities) was conducted by means of PCR, Southern and DNA sequence analyses. Three primer pairs were developed that identified six of the above-mentioned mutations and allowed their molecular description, providing a useful tool for further germplasm screening or marker assisted progeny selection in breeding programs involving the newly identified material. We have found that a complete gene deletion is responsible for a null allele at the Wx-B1 locus in one bread wheat line, whereas sequencing of the corresponding fragments showed a 724 bp deletion in the Wx-D1 locus in one line of bread wheat and an insertion of 89 bp in the Wx-A1 locus in one line of durum wheat, respectively. In addition, nucleotide substitutions and various insertions/deletions ranging from 3 to 30 bp were detected in the PCR fragments of one durum wheat line with a Wx-B1 protein with a different electrophoretic mobility. A fourth primer set, specific for this mutation, was consequently derived. PMID- 15834697 TI - The Vrn-H2 locus is a major determinant of flowering time in a facultative x winter growth habit barley (Hordeum vulgare L.) mapping population. AB - With the aim of dissecting the genetic determinants of flowering time, vernalization response, and photoperiod sensitivity, we mapped the candidate genes for Vrn-H2 and Vrn-H1 in a facultative x winter barley mapping population and determined their relationships with flowering time and vernalization via QTL analysis. The Vrn-H2 candidate ZCCT-H genes were completely missing from the facultative parent and present in the winter barley parent. This gene was the major determinant of flowering time under long photoperiods in controlled environment experiments, irrespective of vernalization, and under spring-sown field experiments. It was the sole determinant of vernalization response, but the effect of the deletion was modulated by photoperiods when the vernalization requirement was fulfilled. There was no effect under short photoperiods. The Vrn H1 candidate gene (HvBM5A) was mapped based on a microsatellite polymorphism we identified in the promoter of this gene. Otherwise, the HvBM5A alleles for the two parents were identical. Therefore, the significant flowering time QTL effect associated with this locus suggests tight linkage rather than pleiotropy. This QTL effect was smaller in magnitude than those associated with the Vrn-H2 locus and was significant in two-way interactions with Vrn-H2. The Vrn-H1 locus had no effect on vernalization response. Our results support the Vrn-H2/Vrn-H1 repressor/structural gene model for vernalization response in barley and suggest that photoperiod may also affect the Vrn genes or tightly linked loci. PMID- 15834698 TI - Genetic analysis of citrus leafminer susceptibility. AB - Damage caused by the citrus leafminer (CLM), Phyllocnistis citrella, is highly dependent on the citrus flushing pattern. Chemical control is only required in young trees, both in nurseries and in newly established orchards. However, this situation is completely different in countries where the causal agent of citrus canker, the bacterium Xanthomonas axonopodis pv. citri exists. CLM infestation results in a higher incidence of citrus canker infection. Among preventive control strategies that provide environmentally sound and sustainable solutions, resistant or tolerant varieties remain the most economical means of insect control. The objective of the present study is to genetically analyse the resistance/susceptibility to CLM and two other traits that might be related, the deciduous behaviour and leaf area of the tree, in a progeny of citradias derived from the cross between two species with different CLM susceptibility--C. aurantium L. and Poncirus trifoliata--using linkage maps of each parent that include several resistance gene analogues. We detected two antibiosis and six antixenosis putative quantitative trait loci (QTLs) in a random sample of forty two of those citradias. An important antibiosis QTL (R2=18.8-26.7%) affecting both percentage of infested leaves and number of pupal casts per leaf has been detected in P. trifoliata linkage group Pa7, which is in agreement with the CLM antibiotic character shown by this species, and independent from any segregating QTL involved in its deciduous behaviour. The maximum value for the Kruskal-Wallis statistic of the other putative antibiosis QTL coincides with marker S2-AS4_800 in sour orange linkage map. Given that the sequence of this marker is highly similar to several nucleotide binding site-leucine-rich repeat (NBS-LRR)-type resistance genes, it might be considered as a candidate gene for insect resistance in citrus. PMID- 15834699 TI - Construction and characterization of BAC libraries from major grapevine cultivars. AB - Genome projects were initiated on grapevine (Vitis vinifera L., 2n=38, genome size 475 Mb) through the successful construction of four bacterial artificial chromosome (BAC) libraries from three major cultivars, Cabernet Sauvignon (Cabernet S), Syrah and two different clones of Pinot Noir (Pinot N). Depending on the library, the genome coverage represented 4.5-14.8 genome equivalents with clones having a mean insert size of 93-158 kb. BAC pools suitable for PCR screening were constructed for two of these BAC libraries [Cabernet S and Pinot N clone (cl) 115] and subsequently used to confirm the genome coverage of both libraries by PCR anchoring of 74 genetic markers sampled from the 19 linkage groups. For ten of these markers, two bands on separate BAC pools were differentiated that could correspond either to different alleles or to a duplication of the locus being studied. Finally, a preliminary assessment of the correspondence between genetic and physical distances was made through the anchoring of all the markers mapped along linkage group 1 of the V. vinifera genetic map. A pair of markers, 2.1 cM apart, anchored the same BAC clones, which allowed us to estimate that 1 cM corresponded in this particular region to a maximum length of 130 kb. PMID- 15834700 TI - No association between insulin gene variation and adult metabolic phenotypes in a large Finnish birth cohort. AB - AIMS/HYPOTHESIS: Although the variable number tandem repeat (VNTR) minisatellite 5' to the insulin gene is among the most studied polymorphisms in diabetes, the relationships between VNTR variation, diabetes-related traits and predisposition to type 2 diabetes remain unclear. Since inadequate sample size is likely to have been an obstacle to reliable inference, we examined the relationship between VNTR variation and a range of diabetes-related traits in a cohort of 5,753 Finnish adults. MATERIALS AND METHODS: VNTR genotypes were derived, by typing at the 23HphI variant site, for 5,646 individuals from the Northern Finland Birth Cohort 1966. Associations were sought between these genotypes and a range of anthropometric (BMI, WHR), physiological (blood pressure) and biochemical (fasting glucose, insulin, lipids, indices of insulin sensitivity and beta cell function) measures obtained at clinical examination at 31 years. RESULTS: We found no evidence that VNTR genotype was significantly associated with measures of insulin secretion, insulin sensitivity, glycaemia, adiposity or blood pressure. CONCLUSIONS/INTERPRETATION: Despite evidence from several relatively small studies suggesting that INS-VNTR genotypes are associated with predisposition to type 2 diabetes, reduced beta cell function and measures of adiposity, the present study failed to detect any association with a range of diabetes-related traits. Taken with other recent studies in large population based cohorts, these data suggest that previous studies have, at the very least, overestimated the influence of the INS-VNTR on type 2 diabetes-related traits. The effects of INS-VNTR variation on insulin transcription observed in vitro appear not to translate into detectable differences in basal insulin secretion in humans. PMID- 15834701 TI - Dynamic changes of GAD65 autoantibody epitope specificities in individuals at risk of developing type 1 diabetes. AB - AIMS/HYPOTHESIS: Progression to type 1 diabetes is associated with intramolecular epitope spreading to disease-specific antibody epitopes located in the middle region of glutamic acid decarboxylase 65 (GAD65). METHODS: The relationship between intramolecular epitope spreading of autoantibodies specific to GAD65 in relation to the risk of developing type 1 diabetes was tested in 22 high-risk individuals and 38 low-risk individuals. We determined the conformational epitopes in this longitudinal study by means of competition experiments using recombinant Fab of four GAD65-specific monoclonal antibodies. RESULTS: Sera from high-risk children in the preclinical stage recognise a specific combination of GAD65 antibody epitopes located in the middle and the C-terminus of GAD65. High risk of progressing to disease is associated with the emergence of antibodies specific for conformational epitopes at the N-terminus and the middle region. Binding to already established antibody epitopes located in the middle and at the N-terminus increases and shows a significant relation (p=0.005) with HLA, which confers risk of developing diabetes. CONCLUSIONS/INTERPRETATION: In type 1 diabetes, GAD65 antibodies are initially generated against the middle and C terminal regions of GAD65. In genetically predisposed subjects the autoimmune response may then undergo intramolecular epitope spreading towards epitopes on the N-terminus and further epitopes located in the middle. These findings clearly demonstrate that the GAD65 autoantibody response in the preclinical stage of type 1 diabetes is dynamic and related to the HLA genotypes that confer risk of diabetes. GAD65-specific Fab should prove useful in predicting progression from islet autoimmunity to clinical onset of type 1 diabetes. PMID- 15834702 TI - [Early results of one-stage septic revision arthroplasties with antibiotic-laden cement. A clinical and statistical analysis]. AB - BACKGROUND: Periprosthetic infections will generally require revision arthroplasty. The one-stage revision arthroplasty with antibiotic-laden cement is hence an attractive therapeutic option, since it only requires one operation, has a low morbidity and, if successful, is cost-efficient. MATERIALS AND METHODS: We performed one-stage revision arthroplasties. The exchanged prostheses were fixed with antibiotic-laden cement after biotic resistance was determined. All patients were treated with postoperative systemic antibiotics. RESULTS: After a mean postoperative examination period of 18.4 months, we confirmed eradication of infection in 14 of 15 knee joints and in 15 of 16 hip joints. The mean duration of hospital stay was 23 days. Patients' satisfaction was high (93.55%) and the clinical results were satisfactory. CONCLUSION: Our analysis shows that our low early reinfection rate (6.45%) is within statistical expectation. Furthermore, we have showed that there is evidence to suggest that the rate of successful outcomes in one-stage revision arthroplasties, at least at the hip, is not different from the rate of two-stage revision arthroplasties and that the difference may be interpreted as stochastic deviation (p=0.264494). According to existing studies, an analogy to knee arthroplasties can be assumed. We demonstrated that one-stage revision is an adequate therapeutic option if patients are carefully selected and under the supervision of an experienced surgeon. PMID- 15834703 TI - Comparative effects of early randomized immune or non-immune-enhancing enteral nutrition on cytokine production in children with septic shock. AB - OBJECTIVE: To compare the effect of early enteral feeding using immune-enhancing (IE) vs. non-immune-enhancing (NIE) formulas on cytokines in children with septic shock. DESIGN AND SETTING: A single-center, randomized, blinded controlled trial in a pediatric intensive care unit of a university hospital. PATIENTS: We randomized 38 patients with septic shock to either IE or NIE. Feedings were advanced to a target volume of energy intake equal to 1/2, 1, 5/4, 6/4, and 6/4 of the predicted basal metabolic rate on days 1-5, respectively. MEASUREMENTS AND RESULTS: Interleukins (IL) 1beta, 6, and 8, tumor necrosis factor alpha, C reactive protein, Pediatric Risk of Mortality (PRISM) score, survival, secondary infections, length of stay, and mechanical ventilation were compared within and between the two groups. Actual mean energy and protein intakes did not differ between the two groups and the caloric-protein balance was not correlated to cytokine levels. On day 5 IL-6 levels were significantly lower (11.8+/-2.4 vs. 38.3+/-3.6) and IL-8 significantly higher in the IE than in the NIE group (65.4+/ 17 vs. 21+/-2.5). After 5 days of nutritional support a significant decrease in IL-6 levels was recorded only in group IE (mean of paired differences 39.4+/-3 pg/ml). In multivariate regression analysis the variation in cytokines was independently correlated only to PRISM (R(2)=-0.50), but pediatric intensive care unit outcome endpoints did not differ between the two groups. CONCLUSIONS: Early IE nutrition may modulate cytokines in children with septic shock, but there is no evidence that this immunomodulation has any impact on short-term outcome. PMID- 15834704 TI - Transcranial Doppler to detect on admission patients at risk for neurological deterioration following mild and moderate brain trauma. AB - OBJECTIVE: To investigate the contribution of transcranial Doppler measurements obtained in the emergency room for detecting patients with secondary neurological deterioration after mild or moderate brain trauma. DESIGN AND SETTING: Prospective cohort study in the emergency room in a university teaching hospital. PATIENTS: Seventy-eight adult patients admitted to the emergency room after a traumatic brain injury (TBI), including 42 patients with Glasgow Coma Score 14-15 and 36 with 9-13. MEASUREMENTS AND RESULTS: All patients had transcranial Doppler measurements on both middle cerebral arteries and computed tomography on admission. Neurological outcome was assessed 7 days after trauma. Of the patients included 7 and 10 had secondary neurological deterioration after mild and moderate TBI, respectively. On admission these groups of patients had significantly more injuries on computed tomography using the Trauma Coma Data Bank classification and higher pulsatility index using transcranial Doppler than the patients having no subsequent neurological worsening. CONCLUSIONS: Increased pulsatility index after mild or moderate TBI is a reason for concern about the possibility of further neurological deterioration. Computed tomography and Doppler measurements could be combined to detect on admission patients at risk for secondary neurological deterioration in order to improve their initial disposition. PMID- 15834705 TI - Admission of patients with severe and moderate traumatic brain injury to specialized ICU facilities: a search for triage criteria. AB - OBJECTIVE: To investigate whether triage for direct admission of patients with traumatic brain injury to a trauma center is facilitated by predicting the risk of potentially removable lesions or raised intracranial pressure (ICP). DESIGN AND SETTING: Cohort study in a level I university trauma center. PATIENTS AND PARTICIPANTS: A prospective cohort of primarily (n=200) and secondarily (n=75) referred patients with moderate or severe traumatic brain injury. MEASUREMENTS AND RESULTS: Predictive characteristics for the risk of surgically removable lesions and the risk of raised ICP (repeatedly > or = 20 mmHg) were identified and included in prognostic models. These models were validated internally with bootstrapping techniques and externally on a historic sample (n=205) regarding discriminative ability (AUC). Among the cohort patients, 67% had raised ICP and 54% had surgically removable lesions. Both outcomes occurred more frequently in patients secondarily referred, but the incidence in patients primarily referred was also high (62% and 33% respectively). No strong predictors of raised ICP were identified. Age and pupillary reactivity were significant predictors of surgically removable lesions. The models discriminated reasonably for surgically removable lesions (AUC=0.78 at development and AUC=0.67 at external validation) but not for raised ICP (AUC=0.59 at development and AUC=0.50 at external validation). CONCLUSIONS: It is difficult accurately to identify patients in need of specialized intensive care using baseline characteristics. The high incidence of both outcomes in patients primarily referred support direct admission of more and particularly older patients with severe or moderate brain trauma to level I trauma centers. PMID- 15834706 TI - Intrahospital transport of a patient with acute exacerbation of chronic obstructive pulmonary disease under noninvasive ventilation. PMID- 15834707 TI - Indwelling time and risk of infection of dialysis catheters in critically ill cancer patients. AB - OBJECTIVE: Despite the lack of evidence to support routine scheduled replacement of dialysis catheters (DCs) this practice continues to be widely used in many intensive care units (ICUs). This study evaluated whether additional risks of catheter-related infection (CRI) are incurred with a conservative attitude in critically ill cancer patients. DESIGN AND SETTING: Prospective, observational study over a 14-month period in a 15-bed medicosurgical unit in a comprehensive cancer center. PATIENTS: Seventy-nine double-lumen DCs were evaluated in 47 patients. Incidence rates of infection per 1000 days of catheter use were examined over 7-day periods. MEASUREMENTS AND RESULTS: The mean indwelling time was 6.9+/-5.5 days. Twelve DCs (15.2%) were removed for suspected CRI. Catheter tip cultures remained negative in 74 cases (93.7%). Overall, one bacteremic CRI, two colonization episodes, and two contaminations were diagnosed, leading to DC colonization and DC-related bacteremia incidence rates of, respectively, 5.4 and 1.8 per 1000 days. When the catheter colonization rate was examined at 7-day intervals, the incidence rate was similar whatever the indwelling time: 5.8, 4.8, and 6.0 per 1000 days, respectively, for the 49 catheters left in place for 7 days or less, 8-14 days (21 DCs), and more than 14 days (9 DCs). The DC colonization incidence rate was similar to that of the 42 short-term catheters inserted during the same period in the same patients (5.9 per 1000 days). CONCLUSIONS: The stable low risk for DC-related infections over time does not support the rationale for scheduled replacement, even in immunocompromised cancer patients. PMID- 15834708 TI - hOGG1 SER326CYS genetic polymorphism in a Turkish population. AB - Oxidative DNA damage, caused by either endogenous or exogenous sources of reactive oxygen species (ROS), has been linked to aging, chronic degenerative diseases, inflammatory diseases and cancers. 8-Hydroxydeoxyguanine (8-OHdG) is a major lesion produced by ROS. Among various types of DNA base modifications, 8 OHdG has been the most widely studied and is considered a key biomarker of oxidative DNA damage. Human 8-oxoguanine DNA glycosylase 1 (hOGG1) is a key component of the base excision repair (BER) pathway and catalyzes the removal of 8-OHdG. Ethnic and inter-individual differences in hOGG1 activity and several kinds of polymorphisms at the hOOG1 gene locus have been observed in the different populations studied so far. Since no information is available on the inter-individual variability of the hOGG1 genotype in the Turkish population, we genotyped 206 healthy, unrelated Turkish individuals. The allelic frequencies of the hOGG1 gene in the Turkish population were found to be 0.50, 0.41 and 0.09 (Ser/Ser, Ser/Cys and Cys/Cys, respectively). Our results are similar to those for Caucasians studied previously but are different from Asian populations. It seems that there is a growing need for extensive genotype studies with respect to the hOGG1 gene due to its importance to various types of cancer and to smoking habits. PMID- 15834710 TI - Temporal constraints on interactions across kinaesthetic channels. AB - We investigated the effect of correlation in background noise on the ability of participants to detect a kinaesthetic target on the index fingers of their hands. Participants had to judge whether the target (a smoothed ramp with quarter-sine onset and offset), was on the left or right finger (experiments 1 and 2). These targets were embedded in noise generated by pseudo-random up-down movements of both the left and right fingers. Positive correlation between the noise on the left and right fingers aided discrimination of the target signal relative to when the noise was uncorrelated. However, this benefit of positive correlation was subject to temporal limitations and diminished with increasing lag between the noise on the fingers (experiment 2). Tests of explicit detection of correlation in kinaesthetic noise (experiment 3) showed a similar pattern, with detection of correlation decreasing with the temporal lag between the noise on the fingers. The results suggest that kinaesthetic signal detection involves sensory integration across the fingers within a limited temporal window. PMID- 15834711 TI - Adaptation to a novel multi-force environment. AB - Humans display accurate limb behavior when they move in familiar environments composed of many simultaneously-acting forces. Little is known about how multi force environments are represented and whether this process partitions between the underlying force components, reflects the net forces present, or is cued to the force-context. We tested between these three main alternatives by examining how reaching movements adapt to a novel multi-force field composed of a velocity dependent force and a constant force. These hypotheses were dissociated first by making the constant force larger and oppositely-oriented to the velocity dependent force; thereby, the net force was always opposite the velocity dependent component. Second, we tested adaptation with all novel forces removed to eliminate any potential cues for the force-context. In two experiments that used forces perpendicular or parallel to the forward movement direction, we found adaptation aftereffects consistent with a mechanism that partitioned the velocity dependent component from the net force field. Specifically, we found aftereffects opposite the rightward or resistive velocity-dependent component of the multi force field, even though the net force imposed was leftward or assistive, respectively. An additional experiment suggested that the velocity-dependent component is partitioned relative to the background load in a limb-based coordinate frame. PMID- 15834712 TI - Magnetic resonance imaging in 120 patients with intractable partial seizures: a preoperative assessment. AB - The aim of this study was to describe magnetic resonance imaging (MRI) findings in patients with medically intractable epilepsy and to compare different magnetic resonance (MR) sequences in order to establish a dedicated and shorter scan time imaging protocol of choice. One hundred and twenty patients with seizures that were refractory to medical treatment were assessed by MRI with spin-echo (SE) T1, fast spin-echo (FSE) T2, fluid-attenuated inversion recovery (FLAIR), inversion recovery (IR) and contrast-enhanced T1 SE sequences. Pathological scans were acquired in 78 patients. Hippocampal sclerosis was detected in 30 patients (25%), cerebral, tumoral, mass lesions in 12 patients (10%), vascular malformations in nine patients (7.5%), cortical infarcts in eight patients (6.7%), cerebral infections in four patients (4.2%) and developmental disorders in 15 patients (12.5%). The most common location of the lesions was the temporal lobe (60%). Coronal, thin (slice thickness 4-5 mm) images have proven to be the most useful in the assessment of the hippocampus. FLAIR and IR are particularly useful in the detection of lesions abutting cerebrospinal fluid (CSF) spaces and developmental disorders, respectively, while T1 SE sequences before and after the intravenous administration of gadolinium offer great facility in identifying space-occupying lesions and infections. MRI is the most important diagnostic tool for the assessment of epileptogenic foci, thus playing the primary role in indicating the type of treatment to be applied. PMID- 15834713 TI - Molecular cloning of the cDNA encoding laccase from Trametes versicolor and heterologous expression in Pichia methanolica. AB - A cDNA encoding for laccase was isolated from the ligninolytic fungus Trametes versicolor by RNA-PCR. The cDNA corresponds to the gene Lcc1, which encodes a laccase isoenzyme of 498 amino acid residues preceded by a 22-residue signal peptide. The Lcc1 cDNA was cloned into the vectors pMETA and pMETalphaA and expressed in Pichia methanolica. The laccase activity obtained with the Saccharomyces cerevisiae alpha-factor signal peptide was found to be twofold higher than that obtained with the native secretion signal peptide. The extracellular laccase activity in recombinants with the alpha-factor signal peptide was 9.79 U ml(-1). The presence of 0.2 mM copper was necessary for optimal activity of laccase. The expression level was favoured by lower cultivation temperature. The identity of the recombinant protein was further confirmed by immunodetection using Western blot analysis. As expected, the molecular mass of the mature laccase was 64.0 kDa, similar to that of the native form. PMID- 15834714 TI - Polyglutamic acid (PGA) production by Bacillus sp. SAB-26: application of Plackett-Burman experimental design to evaluate culture requirements. AB - A locally isolated thermostable Bacillus strain producing polyglutamic acid (PGA) was characterized and identified based on 16S rRNA sequencing. Phylogenetic analysis revealed its closeness to Bacillus licheniformis. To evaluate the effect of different culture conditions on the production of PGA, Plackett-Burman factorial design was carried out. Fifteen variables were examined for their significance on PGA production. Among those variables, K(2)HPO(4), KH(2)PO(4), (NH(4))(2)SO(4) and casein hydrolysate were found to be the most significant variables that encourage PGA production. A correlation between cellular growth, PGA and the produced traces of polysaccharides was illustrated. An inverse relationship practice between cell dry weights and the produced PGA was demonstrated. On the other hand, a direct proportional relation was shown between polysaccharides on one side and cell dry weight and produced PGA on the other. The pre-optimized medium, based on statistical analysis, showed a production of 33.5 g/l PGA, which is more than three times the basal medium. PMID- 15834715 TI - Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals. AB - There has been a rapid increase in the number and demand for approved biopharmaceuticals produced from animal cell culture processes over the last few years. In part, this has been due to the efficacy of several humanized monoclonal antibodies that are required at large doses for therapeutic use. There have also been several identifiable advances in animal cell technology that has enabled efficient biomanufacture of these products. Gene vector systems allow high specific protein expression and some minimize the undesirable process of gene silencing that may occur in prolonged culture. Characterization of cellular metabolism and physiology has enabled the design of fed-batch and perfusion bioreactor processes that has allowed a significant improvement in product yield, some of which are now approaching 5 g/L. Many of these processes are now being designed in serum-free and animal-component-free media to ensure that products are not contaminated with the adventitious agents found in bovine serum. There are several areas that can be identified that could lead to further improvement in cell culture systems. This includes the down-regulation of apoptosis to enable prolonged cell survival under potentially adverse conditions. The characterization of the critical parameters of glycosylation should enable process control to reduce the heterogeneity of glycoforms so that production processes are consistent. Further improvement may also be made by the identification of glycoforms with enhanced biological activity to enhance clinical efficacy. The ability to produce the ever-increasing number of biopharmaceuticals by animal cell culture is dependent on sufficient bioreactor capacity in the industry. A recent shortfall in available worldwide culture capacity has encouraged commercial activity in contract manufacturing operations. However, some analysts indicate that this still may not be enough and that future manufacturing demand may exceed production capacity as the number of approved biotherapeutics increases. PMID- 15834716 TI - Cre/loxP-mediated deletion system for large genome rearrangements in Corynebacterium glutamicum. AB - Genome rearrangement is an increasingly important technique to facilitate the understanding of genome functions. A Cre/loxP-mediated deletion system for large scale genome rearrangements in Corynebacterium glutamicum was developed. By comparative analysis of C. glutamicum R and C. glutamicum 13032 genomes, distinct 14.5-kb and 56-kb regions not essential for cell survival were identified and targeted for deletion. By homologous recombination, loxP sites were integrated at each end of the target region. Deletions between the two chromosomal loxP sites in the presence of Cre recombinase were highly efficient. Accurate deletion was observed in all 96 Cre-expressing strains tested. These deletions represent the largest genomic excisions in C. glutamicum reported to date. Despite the loss of 11 and 58 predicted ORF(s), respectively, upon the deletion of the 14.5-kb and 56 kb regions, the cells still exhibited normal growth under standard laboratory conditions. Based on the precision of its deletion, the Cre/loxP system provides a new, efficient genome rearrangement technique for studying C. glutamicum. PMID- 15834717 TI - A heme tag for in vivo synthesis of artificial cytochromes. AB - A genetic approach is described here that enables the specific covalent attachment of heme via a short C-terminal peptide tag to an otherwise non-heme binding protein. Covalent attachment of heme to the apo-protein is catalysed by the cytochrome c maturation system of Escherichia coli. While its original enzymatic activity is retained, the resulting heme-tagged protein is red, has peroxidase activity and is redox active. The presence or absence of a C-terminal histidine tag results in low-spin heme iron with six- or high-spin heme iron with five coordinate ligands, respectively. The heme tag can be used as a tool for the rational design of artificial c-type cytochromes and metalloenzymes, thereby overcoming previous limitations set by chemical approaches. Moreover, the tag allows direct visualisation of the red fusion protein during purification. PMID- 15834718 TI - Cloning and characterization of a gene coding for a hydrophobin, Fv-hyd1, specifically expressed during fruiting body development in the basidiomycete Flammulina velutipes. AB - Hydrophobin cDNA (fv-hyd1), which is specifically expressed during fruiting body development, was isolated from the basidiomycete Flammulina velutipes by differential display screening. Analysis of the genomic structure of fv-hyd1 revealed an open reading frame (ORF) composed of 363 nucleotides and interrupted by three introns. The deduced amino acid sequence of FV-HYD1 showed a similarity to those of other fungal class I hydrophobins and contained eight cysteine residues highly conserved among hydrophobin proteins. The pattern of the hydropathy plot of FV-HYD1 was similar to those of class I hydrophobins. Southern blot analysis of genomic DNA showed that fv-hyd1 existed as a single copy. Northern blot analysis indicated that the fv-hyd1 transcript was not present in vegetative mycelia but markedly increased in level at the primordial stage. Moreover, the fv-hyd1 transcript was abundant even at the mature fruiting body stage. This result indicates that fv-hyd1 could encode a hydrophobin closely associated with fruiting body development. PMID- 15834719 TI - Aerobic induction of respiro-fermentative growth by decreasing oxygen tensions in the respiratory yeast Pichia stipitis. AB - The fermentative and respiratory metabolism of Pichia stipitis wild-type strain CBS 5774 and the derived auxotrophic transformation recipient PJH53 trp5-10 his3 1 were examined in differentially oxygenated glucose cultures in the hermetically sealed Sensomat system. There was a good agreement of the kinetics of gas metabolism, growth, ethanol formation and glucose utilisation, proving the suitability of the Sensomat system for rapid and inexpensive investigation of strains and mutants for their respiratory and fermentative metabolism. Our study revealed respiro-fermentative growth by the wild-type strain, although the cultures were not oxygen-limited. The induction of respiro-fermentative behaviour was obviously due to the decrease in oxygen tension but not falling below a threshold of oxygen tension. The responses differed depending on the velocity of the decrease in oxygen tension. At high oxygenation (slow decrease in oxygen tension), ethanol production was induced but glucose uptake was not influenced. At low oxygenation, glucose uptake and ethanol formation increased during the first hours of cultivation. The transformation recipient PJH53 most probably carries a mutation that influences the response to a slow decrease in oxygen tension, since almost no ethanol formation was found under these conditions. PMID- 15834720 TI - Redirection of metabolism during nutrient feeding in fed-batch cultures of Bacillus thuringiensis. AB - During sporulation, Bacillus thuringiensis produces insecticidal crystal inclusions (Cry proteins) encoded by cry genes. In fed-batch cultures (FBCs), spores and Cry protein yields are usually low, so we therefore studied the pattern of metabolic changes occurring in batch cultures and FBCs of a B. thuringiensis strain having a cry1Aa promoter-lacZ fusion, and their effect on sporulation and cry1A gene expression. In FBCs, there was a redirection of bacterial metabolism and a reduction in the specific growth rate during feeding, even when the nutrient concentration was higher than at the beginning of batch culture. These physiological changes suggest that the transition state is set up during feeding and this set-up seems to have a negative effect on both sporulation and cry1Aa expression. When the filtrate of a culture in the transition state was added to a batch culture early in the first exponential growth phase, it delayed sporulation and cry1Aa expression, thus suggesting that a soluble cellular factor that blocked sporulation might be excreted during the transition state. Citrate production usually started during the transition state but, when a medium rich in free amino acids was fed, citrate was produced from the first growth phase and sporulation was nearly blocked. PMID- 15834721 TI - Isolation and characterization of autotrophic, hydrogen-utilizing, perchlorate reducing bacteria. AB - Recent studies have shown that perchlorate (ClO(4) (-)) can be degraded by some pure-culture and mixed-culture bacteria with the addition of hydrogen. This paper describes the isolation of two hydrogen-utilizing perchlorate-degrading bacteria capable of using inorganic carbon for growth. These autotrophic bacteria are within the genus Dechloromonas and are the first Dechloromonas species that are microaerophilic and incapable of growth at atmospheric oxygen concentrations. Dechloromonas sp. JDS5 and Dechloromonas sp. JDS6 are the first perchlorate degrading autotrophs isolated from a perchlorate-contaminated site. Measured hydrogen thresholds were higher than for other environmentally significant, hydrogen-utilizing, anaerobic bacteria (e.g., halorespirers). The chlorite dismutase activity of these bacteria was greater for autotrophically grown cells than for cells grown heterotrophically on lactate. These bacteria used fumarate as an alternate electron acceptor, which is the first report of growth on an organic electron acceptor by perchlorate-reducing bacteria. PMID- 15834722 TI - Lower extremity compartmental anatomy: clinical relevance to radiologists. AB - A thorough understanding of compartmental anatomy is necessary for the radiologist participating in the care of a patient with a lower extremity musculoskeletal malignancy. Localization of tumor to compartment of origin and identification of extracompartmental spread preoperatively are needed to correctly stage a tumor and determine the appropriate surgical management. An understanding of the locations of fascial boundaries, extracompartmental tissues, and neurovascular structures of the thigh and lower leg facilitates this diagnostic process. For the radiologist planning to biopsy a suspicious musculoskeletal lesion, consultation with the referring orthopaedic surgeon is recommended in order to jointly select an appropriate percutaneous biopsy approach. Adequate preprocedural planning ensures selection of an approach which prevents iatrogenic tumor spread beyond the compartment of origin, protects neurovascular structures, and allows complete resection of the biopsy tract and scar at the time of surgical resection without jeopardizing a potential limb sparing procedure. Cross-sectional anatomic review and case examples demonstrate the importance of a detailed understanding of compartmental anatomy when approaching the patient with a lower extremity musculoskeletal tumor. PMID- 15834723 TI - Cellular immunotherapy: antigen recognition is just the beginning. AB - Advances in molecular and cellular biology have illustrated both the flexibility and complexity involved in host immune responses. Understanding this response is vital to the further development of therapeutic strategies that involve manipulation of the cellular immune response to target tumors. Mobilized, tumor antigen-specific T cells, the core for most immunotherapeutic strategies, are highly regulated, and capable of a wide spectrum of functional responses. Due to differences in murine and human immunity, broad-scale immune monitoring, particularly high-throughput ex vivo analysis of human immune responses, promises to determine what comprises an effective immunotherapy. Such understanding will lead to more sophisticated clinical trials, earlier determination of efficacy and individualized protocols. PMID- 15834724 TI - Surface lesions of the bones of the hand. AB - Surface lesions involving the bones of the hand are uncommon. This pictorial review illustrates the spectrum of conditions including benign primary bone tumours, malignant primary bone tumours and non-neoplastic disorders. The review focuses on the radiographic appearances of these lesions and other techniques such as CT and MR imaging that may suggest a specific diagnosis. PMID- 15834725 TI - [Neurodermatitis. A risk factor for the development of squamous cell carcinoma]. AB - Squamous cell conjunctival carcinomas (SCCC) are rare. We report on a 34-year-old patient with neurodermatitis and chronic conjunctivitis who presented with a therapy-resistant corneal ulcer induced by mechanical irritation diagnosed as SCCC. Chronic blepharoconjunctivitis seems to sustain the development of SCCC in patients with neurodermatitis because of their insufficient immune response. PMID- 15834726 TI - Radioisotope shuntograms at the Children's Hospital of Eastern Ontario. AB - INTRODUCTION: Nuclear medicine shuntograms using the radiotracer technetium-99m diethylenetriaminepentaacetic acid have been used for many years as an additional method to assess shunt patency and performance. METHODS: The medical records of all children who had shuntograms performed at the Children's Hospital of Eastern Ontario between January 13, 2000, and January 10, 2003, were retrospectively reviewed. There was a total of 68 procedures in 56 patients with an average age of 9 years 3 months. During the same period there were 123 shunt revisions. RESULTS: Forty-two shuntograms were reported as normal. Of these, ten were identified to be false negative. Two children with fractured shunts had shuntograms performed in order to assess for cerebrospinal fluid flow patency; the shuntograms were identified to be normal, confirming shunt dependence, and the shunt replaced. Twenty-six shuntograms were reported as abnormal in 24 patients; 17 went on to have surgery and the shunt malfunction was confirmed. Seven patients did not require surgery: five were declared shunt independent, two patients clinically improved after severe constipation was treated. CONCLUSIONS: Approximately one fourth of all shuntograms reported as normal are not (false negative rate=25%). Review of five other major studies between 1980 and 2003 have reported false-negative rates between 2 and 36%, which may be explained by variations in shuntogram protocols. A standardized method is proposed. PMID- 15834727 TI - Intracranial aneurysms in children aged under 15 years: review of 59 consecutive children with 75 aneurysms. AB - OBJECTIVE: The objective was to review the clinical aspects and therapeutic strategies in a series of aneurysmal vasculopathies seen in children 15 years or under. METHODS: From our dedicated neurovascular databank of patients, we reviewed 59 consecutive children who had 75 separate lesions. RESULTS: The children were divided into four age groups: below 2 years (22%), 2-5 years (24%), 6-10 years (24%) and 11-15 years (30%). Thirty-three children had dissecting aneurysms, 2 had chronic post-traumatic aneurysms, 8 had infectious aneurysms and 16 had saccular lesions. Twenty-seven percent of the lesions were in the posterior circulation, and 21% developed on the middle cerebral artery. Most dissecting lesions were encountered in the vertebrobasilar system, while saccular lesions were present mostly in the anterior circulation. Half of all cases presented with haemorrhage. Haemorrhage in patients below 2 years of age was due to dissecting aneurysms, while saccular aneurysms were responsible for haemorrhage in patients above 5 years of age. Five children had familial disease and 9 presented with multiple aneurysms. Forty-eight children were referred to us for treatment. Thirty-two underwent surgical (21.9%), endovascular (62.8%) or combined (9.3%) treatment. Eleven patients were treated conservatively and in 5 patients the aneurysms had spontaneously thrombosed at admission. Overall, complete or partial spontaneous thrombosis was seen in 10 patients (16.9%). Dissecting aneurysms were frequent in children of all ages with either associated thrombosis or arterial tear with repeated acute haemorrhage and poor outcome. Two types of dissection seem identifiable despite the small number of cases collected: acute segmental arterial tear without thrombosis, acute subarachnoid haemorrhage (SAH) and recurrence before 5 years; and subacute focal dissection with partial thrombosis (or mural haematoma), rare SAH and no early recurrence. The former would require aggressive management whereas the latter often do not require interventional approaches. The mortality in our series of aneurysms is low in the treated group (10.42%). The overall tolerance to haemorrhage seems better than in adults, as already stressed in the literature. CONCLUSION: The multiple etiologies encountered confirm the heterogenous nature of "aneurysms". The variety of treatments used suggests the need to categorise aneurysms into subgroups in sufficient numbers to fully appreciate the behavior of the lesions and make the appropriate therapeutic decisions. PMID- 15834728 TI - Venous hypertension and craniosynostosis. AB - INTRODUCTION: This article addresses the relevance of venous hypertension to children born with, in particular, the "syndromic" forms of craniosynostosis. The development of the human cerebral venous system is summarised and the patterns of anomalous venous drainage seen in children with complex forms of craniosynostosis associated with raised intracranial pressure are described. AETIOLOGY: The aetiology of these abnormal patterns is discussed under the general headings of "The constriction theory", "A primary role for FGFR mutations" and "Persistence of the foetal pattern of intracranial venous drainage."Venous hypertension (along with hydrocephalus, cranio-cerebral disproportion and airway obstruction) has an important role in the aetiology of raised intracranial pressure in severely affected children. TREATMENT: The inter-relationship between these factors is explained and then the treatment of those children whose raised intracranial pressure is considered to be due predominantly to venous hypertension is described. PMID- 15834729 TI - Prepubertal unilateral spermatic vessel ligation decreases haploid cell population of ipsilateral testis postpubertally in rats. AB - DNA flow cytometry is a sensitive and rapid technique to evaluate the germ cell maturation of testis. The haploid cell population of testis specifically denotes testicular maturation. High ligation of spermatic vessels (HLSV) and low ligation of spermatic vessels (LLSV) have been described in the surgical treatment of high undescended testis. Estimation of germ cell population of testis after these maneuvers has not been performed so far by DNA flow cytometry; hence this study was designed. Thirty male rats, aged 30 days, were randomized into three groups. Group I underwent sham surgery; group II underwent unilateral HLSV-ligation and division of main spermatic vessels, similar to the Fowler-Stephens technique; and group III underwent unilateral LLSV-ligation and division of main spermatic vessels close to the testis, similar to the Koff and Sethi technique. Thirty days later, the ipsilateral testes were harvested, and haploid, diploid, and tetraploid cells were counted by DNA flow cytometry. The mean (+/-SD) percentage of the haploid cell population was 68.7+/-4.8, 49.3+/-6.5, and 50.8+/-6.4 in groups I, II, and III, respectively. There was a significant (p<0.05) decrease in haploid cell population between groups I and II and between groups I and III. However, there was no significant difference between groups II and III. In conclusion, prepubertal HLSV as well as LLSV decreases the haploid cell population of ipsilateral testis in rats. LLSV has no advantage over HLSV. PMID- 15834731 TI - A papillary thyroid carcinoma case associated with hereditary colon carcinoma due to familial adenomatous polyposis with no hereditary mutation finding. PMID- 15834730 TI - Modulation of immune cell function by polyunsaturated fatty acids. AB - The n-3 and n-6 polyunsaturated fatty acids (PUFAs) are essential dietary constituents. They are important as a source of energy, as structural components of cell membranes, and as signalling molecules. They have been demonstrated to be potent modulators of the immune response, and research has endeavoured to optimise the ratio of n-3 to n-6 PUFAs in the lipid component of total parenteral nutrition (TPN) to optimise their beneficial effects in the clinical setting. Critically ill neonates on TPN have an increased incidence of sepsis, and additional studies have determined that lipid emulsions depress various elements of cellular immune responses in monocytes, lymphocytes, and neutrophils. It has been proposed that PUFAs may mediate their manifold effects through the modification of eicosanoid production and by directly or indirectly modifying intracellular signal transduction pathways, including the alteration of gene transcription, in various tissues. They are susceptible to lipid peroxidation, and there is evidence that the products of this process may result in cell death by apoptosis, a nonphlogistic homeostatic process of cell deletion. PUFAs have been shown to induce apoptosis in primary lymphocytes, colonic mucosal cells, and various cell lines. Additionally, our laboratory has shown them to be potent inducers of apoptosis in neonatal monocytes. This may represent a novel mechanism whereby PUFAs may modify the immune response. PMID- 15834732 TI - Klotho insufficiency causes decrease of ribosomal RNA gene transcription activity, cytoplasmic RNA and rough ER in the spinal anterior horn cells. AB - The klotho gene was identified in 1997 as the gene whose severe insufficiency (kl/kl) causes a syndrome resembling human aging, such as osteoporosis, arteriosclerosis, gonadal atrophy, emphysema, and short life span in a mouse strain. Regarding the gait disturbance reported in kl/kl mice, the present study examined the spinal cord of kl/kl mice, and revealed decreases in the number of large anterior horn cells (AHCs), the amount of cytoplasmic RNA, the number of ribosomes and rough endoplasmic reticulum (rER), and the activity of ribosomal (r) RNA gene transcription without significant loss of the total number of neurons in the ventral gray matter. Increased immunostaining of phosphorylated neurofilament in the AHCs and of glial fibrillary acidic protein in reactive astrocytes in the anterior horn of kl/kl mice were also observed. On the other hand, the posterior horn was quite well preserved. The results suggest that the kl/kl insufficiency causes atrophy and dysfunction of the spinal AHCs through decreased activity of rRNA gene transcription, which may reduce the amount of cytoplasmic RNA and the number of ribosomes and rER. These findings resemble those found in the spinal cord of patients with classic amyotrophic lateral sclerosis (ALS). The results show that klotho gene insufficiency causes dysfunction of the protein synthesizing system in the AHCs, and might indicate the klotho gene is involved in the pathological mechanism of classic ALS. The kl/kl is a new animal model of AHC degeneration, and may provide clues to understanding the etiology of classic ALS. PMID- 15834733 TI - Regulation of progesterone production in human term trophoblasts in vitro by CRH, ACTH and cortisol (prednisolone). AB - BACKGROUND: In most mammals, onset of labor is accompanied with progesterone withdrawal. In humans, cortisol blockade of progesterone is a possible mechanism involved in the initiation of labor. Therefore, aim of the study was to clarify the effect of CRH, ACTH and cortisol (prednisolone) on the release of progesterone by term trophoblast cells in vitro. METHODS: Cytotrophoblast cells were prepared from human term placentas by standard dispersion of villous tissue followed by a percoll gradient centrifugation step. Trophoblasts were incubated with CRH, ACTH as well as with prednisolone. RESULTS: The release of progesterone is decreased in CRH- and ACTH-treated trophoblast cell cultures compared to untreated trophoblast cells. Addition of prednisolone in varying concentrations leads to an increase of trophoblast progesterone production. CONCLUSIONS: The results suggest that CRH and ACTH directly modulate the endocrine function of trophoblasts in culture by downregulating progesterone production. Prednisolone on the other hand showed a stimulating effect on progesterone production in term trophoblast cells in vitro. Because blockade of progesterone is a possible mechanism involved in initiation of labor, we may speculate that CRH and ACTH are directly involved in the auto- or paracrine regulation of this procedure. PMID- 15834734 TI - Polish population study on Y chromosome haplotypes defined by 18 STR loci. AB - Polymorphism of 18 STR loci specific to the human Y chromosome (DYS19, DYS388, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS426, DYS437, DYS438, DYS439, DYS460, GATA H4.1, DYS385 a/b, and YCAII a/b) was evaluated by means of a multiplex (octadecaplex) PCR reaction and capillary electrophoresis in a Polish population sample of 208 unrelated males. A total of 192 different haplotypes and 183 unique haplotypes were identified. The observed haplotype diversity was 0.998, while discrimination capacity was 92.3%. DYS389 was shown to be the most valuable in discrimination of similar haplotypes, whereas DYS388, DYS393, DYS426, and DYS438 did not affect the discrimination power of the multiplex. PMID- 15834735 TI - Recommendations for animal DNA forensic and identity testing. AB - Genetic analysis in animals has been used for many applications, such as kinship analysis, for determining the sire of an offspring when a female has been exposed to multiple males, determining parentage when an animal switches offspring with another dam, extended lineage reconstruction, estimating inbreeding, identification in breed registries, and speciation. It now also is being used increasingly to characterize animal materials in forensic cases. As such, it is important to operate under a set of minimum guidelines that assures that all service providers have a template to follow for quality practices. None have been delineated for animal genetic identity testing. Based on the model for human DNA forensic analyses, a basic discussion of the issues and guidelines is provided for animal testing to include analytical practices, data evaluation, nomenclature, allele designation, statistics, validation, proficiency testing, lineage markers, casework files, and reporting. These should provide a basis for professional societies and/or working groups to establish more formalized recommendations. PMID- 15834736 TI - Cross-reactivity of the CEDIA buprenorphine assay with opiates: an Austrian phenomenon? AB - When testing the Microgenics CEDIA assay for immunological buprenorphine analysis, cross-reactivity between the buprenorphine reagents and opiates was observed at concentrations higher than 120 mg/l morphine, 320 mg/l methadone, 30 mg/l codeine, 60 mg/l dihydrocodeine and 520 mg/l morphine-3-glucuronide. The cross-reactivity with morphine has the greatest impact on routine screening as opiate maintenance therapy in Austria is also performed with slow-release oral morphine. The use of a second cutoff value of 30 mug/l for urine samples that are (immunologically) positive for opiates is therefore suggested, compared to the cutoff value of 5 microg/l proposed by the manufacturer. PMID- 15834738 TI - Congenital systemic arteriovenous fistula between the distal thoracic aorta and hemiazygos vein in a child. PMID- 15834737 TI - Tissue defect at the gunshot entrance wound: what happens to the skin? AB - To investigate the question what happens to the tissue lost at the entrance wound, experimental studies were performed on composite models consisting of dyed pig skin and gelatin blocks. For the test shots to the skin-gelatin preparations, cartridges calibre .38 spec. with different bullet types (round nose, hollow point, flat nose, truncated cone) were used. In all shots, a multitude of coloured skin particles were macroscopically discernible along the bullet tracks. In addition, small cell aggregations could be demonstrated microscopically even in those sections of the bullet paths which did not show skin fragments visible to the naked eye. The distribution of the skin particles showed certain peculiarities depending on the type of projectile. PMID- 15834740 TI - Chlorhexidine and silver-sulfadiazine coated central venous catheters in haematological patients--a double-blind, randomised, prospective, controlled trial. AB - BACKGROUND: Central venous catheters (CVCs) are essential for the intensive care of patients with haematological illness. Catheter-related infections (CRI) are an important problem in modern medicine, which may lead to life-threatening situations, to prolonged hospitalisation and increased cost. In immunocompromised patients suffering from haemato-oncological diseases, CRI is a significant factor for adverse outcome. Several clinical studies have shown that CVCs coated with antiseptics such as chlorhexidine and silver-sulfadiazine (CHSS) reduce the risk of catheter-related bacteraemia. Most studies, however, were performed on intensive care patients not suffering from chemotherapy-induced immunosuppression. PATIENTS AND METHODS: A prospective double-blind, randomised, controlled trial was performed to investigate the effectiveness of CHSS-coated catheters in haemato-oncological patients. A total number of 184 catheters (median duration of placement, 11 days) were inserted into 184 patients (male 115, female 69), of which 90 were antiseptically coated. After removal, all catheters were investigated for bacterial growth. MAIN RESULTS: Catheters coated with CHSS were effective in reducing the rate of significant bacterial growth on either the tip or subcutaneous segment (26%) compared to control catheters (49%). The incidence of catheter colonisation was also significantly reduced (12% coated vs 33% uncoated). Data obtained show a significant reduction of catheter colonisation in CHSS catheters. There was no significant difference in the incidence of catheter-related bacteraemia (3% coated vs 7% uncoated). However, due to the overall low rate of CRI, we could not observe a significant reduction in the incidence of catheter-related bacteraemia. CONCLUSION: Our data show that the use of CHSS catheters in patients with haematological malignancy reduces the overall risk of catheter colonisation and CRI, although the incidence of catheter related bacteremia was similar in both groups. PMID- 15834741 TI - Another form to fill in! Clients' reflections on the hospice use of questionnaires. AB - GOALS OF WORK: The use of validated tools is increasingly accepted as an unqualified good that is viewed as best practice in supportive care. This article begins to explore the impact of standardized questionnaire use in supportive care by presenting findings from recent qualitative research on clients' perceptions of the use of standardized assessment tools during their hospice experience. PATIENTS AND METHODS: There were two arms to this phenomenological descriptive study: A. Interviews with hospice patients and their carers; B. Interviews with hospice staff. The results from arm A are reported in this article. This involved interviews with ten families (available patient and carer) who had hospice experience with questionnaires and ten families who were cared for without questionnaires. The interviews were audiorecorded, transcribed verbatim, and thematically analysed. MAIN RESULTS: The research presented in this article is seminal work in the area which affirms significant concerns about the use of questionnaires in hospice practice. The evidence indicates the majority of clients dislike the use of questionnaires and points to questionnaire use being a practice built around staff, rather than client, needs. The findings also provide insight into the process of collusion by which hospice workers who are enthusiastic about the use of questionnaires can be led to believe, because of client gratitude, that the process is positive. CONCLUSIONS: Questionnaires should not be seen as an unqualified good, and thus should not be automatically accepted as best practice within hospice or palliative care service provision. PMID- 15834742 TI - Alveolar bone levels in adults as assessed on panoramic radiographs. (I) Prevalence, extent, and severity of even and angular bone loss. AB - There is limited information regarding the prevalence and intraoral distribution of infrabony lesions potentially suitable for regenerative procedures in common patients seeking regular dental care in a dental practice. The aim of the present study was to investigate the prevalence and extent of alveolar bone loss and infrabony defects in randomly selected orthopantomograms of adult patients in different age groups seeking treatment at the dental service of the German Armed Forces. A total of 240 panoramic exposures were available for analysis, 60 in each of the age groups <30, 30-39, 40-49, and >or=50 years of age. At each tooth, distances between the cementoenamel junction or margin of restoration and the alveolar crest as well as the bone level were measured with a calliper to the next 0.1 mm. Whereas virtually no bone loss was present in the youngest age group, a major change in bone level frequency distributions occurred after age 30 years. At age 50 years, about 50% of subjects had considerable bone loss of more than 4 mm at 10% or more sites, and 6 mm or more at about 5% sites. Deep infrabony defects were infrequently found before age 40 years. About 20% of patients at least 50 years of age had radiographic evidence of infrabony pockets of 4 mm or more at not more than 5% of sites. Bone loss was more pronounced in the maxilla, especially at molars. In this predominantly male population, periodontal bone loss gradually increased with age. However, prevalence of infrabony defects was very low. PMID- 15834743 TI - [On the indications for and morbidity of segmental resection of the mandible for squamous cell carcinoma in the lower oral cavity]. AB - BACKGROUND: Segmental resection of the mandibula in oral cancer surgery leads to both functional and aesthetic problems. The decision to preserve or resect the mandible depends on the vicinity of the lesion to the bone. Consequently, based on the rules of safety margins to all planes that are recommended for soft tissues, each lesion that is closer than 10 mm to the mandible needs resection of the bone. PATIENTS AND METHODS: To establish data-based treatment modalities, a retrospective study was initiated and the results from all preoperative staging investigations of 152 patients with intraoral squamous cell carcinoma who underwent continuity or marginal resection of the mandible were evaluated. The histological outcome of the resected bone was compared to the staging results. Functional rehabilitation and long-term follow-up including survival rates were evaluated. The study reports on typical complications following segmental resection such as fracture of the reconstruction plate and demonstrates experiences with secondary microsurgical reconstructive surgery. RESULTS: Mainly in cases of stage T1 and T2 carcinomas which are closer than 10 mm to the bone and clinically do not show any infiltration to the mandible, a marginal resection seems to be adequate. The decision about the extension of mandibular resection can be based on intraoperative cross sectional investigation of the periosteum. The survival rate of patients with intraoral carcinomas close to the mandible who underwent marginal mandibulectomy seems to be the same as in cases of continuity resection. A more conservative management of mandibular resection seems to be adequate and a data-based concept to standardize therapy of mandibular resection is presented. PMID- 15834744 TI - [Assessment of severity of oral lichen planus using a new clinical index]. AB - AIM: The goal of this study was to develop a clinical index to assess therapy of oral lichen planus (OLP) to be used in comparing the efficacy of two topical glucocorticoids. The clinical severity of non-gingival and gingival lichen planus and their severity from the patients' view were evaluated. DESCRIPTION OF THE INDEX: A clinical index for assessing the severity of oral lichen planus is described with which the clinical forms as well as all the varying combinations of forms can be addressed separately. The clinical classification of OLP into six variants according to Andreasen [1] is the basis of the index. The index provides information about the severity of the disease in single regions and in the patient as a whole. The index is subdivided into a basic index as well as supplementary indices, which are optional. The clinical forms and size of OLP are recorded using the basic index, while various form-specific features such as the intensity of whitish pattern or mucosal erythema can be assessed additionally using the supplementary indices. Non-gingival and gingival lichen planus are assigned with their own indices. An index of subjective symptoms consisting of a visual analogue scale and verbal scales is available for describing patient discomfort. There is also a short form of the index (study-region index/one region index), whereby only a single selected region is assessed. CONCLUSION: The index presented here is available as a new method of assessing OLP severity, offering both basic and advanced information for use especially in scientific studies of oral lichen planus. PMID- 15834745 TI - Disseminated histoplasmosis in an HIV-infected patient discovered by routine blood smear staining. PMID- 15834750 TI - Lack of effect of shorter turnaround time of microbiological procedures on clinical outcomes: a randomised controlled trial among hospitalised patients in the Netherlands. AB - Shortening the turnaround time of microbiological procedures was associated with an improved clinical outcome in two studies performed in the USA. To study the clinical impact of a shortened turnaround time in a northwest European setting in which an automated system was used for bacterial identification and susceptibility testing, a single-blind, prospective, randomised controlled trial was conducted in a hospital in the Netherlands. All hospitalised patients with a bacterial infection confirmed by culture were randomly assigned to a control (conventional) group or an intervention (rapid) group. Overnight methods were used for identification and susceptibility testing in the control group, while the Vitek 2 system (bioMerieux, Marcy l'Etoile, France) was used in the rapid group. In each of three consecutive study periods, accelerating factors were added progressively to the laboratory workflow of the rapid group to increase same-day reporting, whereas methods remained identical in the conventional group. The turnaround time of the microbiological cycle using the Vitek 2 system as compared to conventional methods was studied and the clinical impact of a shortened turnaround time assessed in terms of mortality, morbidity, and cost. For the rapid groups, the turnaround time was significantly shorter for oral reporting of final susceptibility results in all three study periods and for reporting on paper in the third study period. There was no significant difference between groups in any of the clinical impact variables. Vitek 2 results were available for reporting significantly earlier as compared to conventional testing. For the overall patient group in our hospital setting, however, this had no clinical impact. PMID- 15834752 TI - Mutations in Sendai virus variant F1-R that correlate with plaque formation in the absence of trypsin. AB - With the emergence of new viruses, such as the SARS virus and the avian influenza virus, the importance of investigations on the genetic basis of viral infections becomes clear. Sendai virus causes a localized respiratory tract infection in rodents, while a mutant, F1-R, causes a systemic infection. It has been suggested that two determinants are responsible for the systemic infection caused by F1-R [Okada et al (1998) Arch Virol 143:2343-2352]. The primary determinant of the pantropism is the enhanced proteolytic cleavability of the fusion (F) protein of F1-R, which allows the virus to undergo multiple rounds of replication in many different organs, whereas wild-type virus can only undergo multiple rounds of replication in the lungs. The enhanced cleavability of F1-R F was previously attributed to an amino acid change at F115 that is adjacent to the cleavage site at amino acid 116. Secondly, wild-type virus buds only from the apical domain of bronchial epithelium, releasing virus into the lumen of the respiratory tract, whereas F1-R buds from both apical and basolateral domains. Thus, virus is released into the basement membrane where it can easily gain access to the bloodstream for dissemination. The microtubule disruption is attributed to two amino acid differences in M protein. To confirm that the F and M gene mutations described above are solely responsible for the phenotypic differences seen in wild-type versus F1-R infections, reverse genetics was used to construct recombinant Sendai viruses with various combinations of the mutations found in the M and F genes of F1-R. Plaque assays were performed with or without trypsin addition. A recombinant virus containing all F1-R M and F mutations formed plaques in LLC-MK2 cells and underwent multiple cycles of replication without trypsin addition. To clarify which mutation(s) are necessary for plaque formation, plaque assays were done using other recombinant viruses. A virus with only the F115 change, which was previously thought to be the only change important for plaque formation of F 1-R F, did not confer upon the virus the ability to form plaques without the addition of trypsin. Another virus with the F115 and both M changes gave the same result. Therefore, more than one mutation in the F gene contributes to the ability of F1-R to form plaques without trypsin addition. PMID- 15834753 TI - Low density lipoprotein inactivates Vibrio vulnificus cytolysin through the oligomerization of toxin monomer. AB - Blood lipoprotein has been shown to be an important defense factor against the bacterial infection. Lipoprotein is scavenger of bacterial endotoxin (lipopolysaccharide, LPS). However, there is little evidence showing its protective action against the bacterial exotoxin. We have previously demonstrated that cholesterol inactivates Vibrio vulnificus cytolysin (VVC) through oligomerization of the toxin monomer. The aim of the present study was to investigate the relationship between VVC and low-density lipoprotein cholesterol (LDL-C). LDL induced the oligomerization of VVC in a dose-dependent manner. The oligomerization of VVC monomer with LDL was demonstrated by immunoblotting, which exhibited 200-kDa bands corresponding to a tetramer of the native VVC of relative molecular weight of 51,000. Moreover, LDL inactivated hemolytic activity of VVC in a dose-dependent manner, and this response was not changed by the modifications of LDL (heat denaturation of proteins and peroxidation of phospholipids), suggesting that LDL-C is associated with the defense action against VVC. These results suggest that LDL-C inactivates VVC through the induction of toxin oligomerization. PMID- 15834754 TI - Impact of the Expanded Program of Immunization against hepatitis B infection in school children in Malaysia. AB - The implementation of the Expanded Program of Immunization (EPI) in 1989 has dramatic impact on hepatitis B virus (HBV) infection in school children in Malaysia. A cross-sectional seroprevalence study of HBV infection in 190,077 school children aged 7-12 years from 1997 to 2003 showed a steady decline of HBV surface antigen (HBsAg) prevalence rate from 2.5% for children born in 1985 to 0.4% among school children born in 1996. The overall prevalence of HBsAg was 0.6%, 0.7% in males and 0.6% in females. Over 92.7% of school children had been vaccinated with HBV vaccine, in which 93.7% were vaccinated under the EPI and 6.3% on voluntary basis. The school children vaccinated under EPI had a 0.4% HBsAg carrier rate, which was significantly lower than school children vaccinated on a voluntary basis (HBsAg carrier rate 1.3%) and non-vaccinated school children (HBsAg carrier rate 2.7%), suggesting that HBV vaccination of infants was the most effective measure in preventing vertical transmission of HBV in the hyperendemic region. PMID- 15834755 TI - Fecundity limits in Frangula alnus (Rhamnaceae) relict populations at the species' southern range margin. AB - The geographic range of many temperate plant species is constrained by climate, but it remains little known how climate affects population performance at low latitude range margins. This study investigated the reproduction of the Eurasian tree Frangula alnus in relict populations near its southwestern range limit in southern Spain. The aim was to identify the principal stages and causes of ovule loss experienced by these marginal populations. More than 6,800 flowers were monitored over 2 years, insect observations and different experiments were carried out to assess levels of pollen and resource limitation, as well as the influence of flowering phenology on seed production. Most ovule losses occurred during flower anthesis and were due to strong cross-pollen limitation. Fruit set was affected by tree size, light regime and flowering phenology, probably through their effects on pollinator behaviour. Fruit set was almost zero throughout the first half of the flowering season. Then it increased paralleling changes in pollinator abundance but was soon overridden by increasing ovule desiccation due to summer drought. Successful seed production was mostly confined to a brief period near the end of the flowering season. Adverse weather during this period in the second study year resulted in a threefold bud and flower mortality and a 50% decrease of fruit production. Spring rainfalls in southwestern Spain have diminished more than 30% through past decades leading to an earlier onset of summer drought. This trend and its adverse effects on seed production may contribute to explaining the recent decline of F. alnus at its southwestern range limit. PMID- 15834756 TI - GLV supplements increased plasma beta-carotene, vitamin C, zinc and hemoglobin in young healthy adults. AB - BACKGROUND: Green leafy vegetables (GLV) are rich sources of beta-carotene, iron and other micronutrients. Our in vitro studies have demonstrated good antioxidant potential in GLV. Moreover linkages of GLV intakes with plasma retinol and ascorbic acid were seen in apparently healthy Indians. AIM OF THE STUDY: To investigate the effect of GLV as a natural fortificant of multiple micronutrients through a prospective human trial. METHODS: Short-term (0-4 h) response (AUC) of single dose of 7.9 mg beta-carotene and 130 mg ascorbic acid through a spinach carrot meal against the standard meal without GLV plus10 mg beta-carotene and 150 mg ascorbic acid tablets was studied in two groups of 4 young volunteers each. In the second trial of 3 weeks' supplementation, 5 groups of young adults (n = 40) were given either 100 g GLV/day alone or with tablets of vitamin E (100 mg/day), or C (100 mg/day) or more oil (5 g/day) or non-GLV meal with tablet of beta carotene (10 mg/day). Hemoglobin (Hb), plasma beta-carotene, zinc, vitamin C, glucose, and triglycerides were measured. RESULTS: In a postprandial response, AUC were comparable in both GLV and standard meals for beta-carotene and ascorbic acid. In case of triglycerides and glucose AUC the GLV meal showed a better recovery to the baseline value after 4 hours than the standard meal. Three weeks' supplementation of GLV with more oil resulted in significant increase of plasma beta-carotene (51%) and Hb (9%). GLV with vitamin E showed a significant increase in plasma beta-carotene (40%), Hb (8%) and plasma vitamin C (6%). Supplementing beta-carotene without GLV significantly increased Hb (11%), plasma zinc (14%) in addition to beta-carotene. Multiple regression analyses weighted for energy intake indicated a significant association of percent increase in Hb with intakes of iron, riboflavin, folic acid, beta-carotene, copper, phytate and fiber (p < 0.01), percent change in plasma zinc with intakes of zinc, beta-carotene, vitamin C, riboflavin, copper, iron, and thiamin (p < 0.01), percent change in vitamin C with intakes of vitamin C, vitamin E, niacin, riboflavin, thiamin, beta-carotene, zinc, phytate and fiber (p < 0.05) and percent change in plasma beta-carotene with intakes of beta-carotene, thiamin, folic acid, zinc, phytate and tannins (p < 0.05). CONCLUSION: Using 100 g GLV/day with 10 g oil could be a single moderate strategy for supplementation of iron, beta-carotene, ascorbic acid and zinc. PMID- 15834757 TI - Stability and biotransformation of various dietary anthocyanins in vitro. AB - BACKGROUND: Anthocyanins, which are found in high concentrations in fruit and vegetable, may play a beneficial role in retarding or reversing the course of chronic degenerative diseases. However, little is known about the biotransformation and the metabolism of anthocyanins so far. AIM OF THE STUDY: The aim of the study was to investigate possible transformation pathways of anthocyanins by human faecal microflora and by rat liver microsomes as a source of cytochrome P450 enzymes as well as of glucuronyltransferases. METHODS: Pure anthocyanins, an aqueous extract of red radish as well as the assumed degradation products were incubated with human faecal suspension. The incubation mixtures were purified by solid-phase extraction and analysed by HPLC/DAD/MS and GC/MS. Quantification was done by the external standard method. Furthermore the biotransformation of anthocyanins by incubation with rat liver microsomes in the presence of the cofactor NADPH (as a model for the phase I oxidation) and in the presence of activated glucuronic acid (as a model for the phase II glucuronidation) was investigated. RESULTS: Glycosylated and acylated anthocyanins were rapidly degraded by the intestinal microflora after anaerobic incubation with a human faecal suspension. The major stable products of anthocyanin degradation are the corresponding phenolic acids derived from the B ring of the anthocyanin skeleton. Anthocyanins were not metabolised by cytochrome P450 enzymes, neither hydroxylated nor demethylated. However they were glucuronidated by rat liver microsomes to several products. CONCLUSIONS: The gut microflora seem to play an important role in the biotransformation of anthocyanins. A rapid degradation could be one major reason for the poor bioavailability of anthocyanins in pharmacokinetic studies described so far in the literature. The formation of phenolic acids as the major stable degradation products gives an important hint to the fate of anthocyanins in vivo. PMID- 15834758 TI - Hypermetabolic pattern in frontal cortex and other brain regions in unmedicated schizophrenia patients. Results from a FDG-PET study. AB - We report results of a FDG-PET study in 10 patients with schizophrenia (6 unmedicated, 4 never medicated) and 12 healthy age-matched controls. The patients met ICD-10 and DSM-IV criteria for schizophrenia and all reported psychotic, "positive" symptoms when tested. Schizophrenic patients had higher absolute CMRGlu values in almost all quantified regions compared to normal subjects. Using the occipital cortex as the reference region patients showed a hyperfrontal metabolic pattern. Other significant regional differences were found with respect to thalamus, striatum and temporal cortex. The finding of a hyperfrontality in un and never medicated psychotic schizophrenic patients must be discussed in the light of the psychopathological symptoms of patients when tested, a possible disruption of cortico-striato-thalamic feedback loops and recent findings of a hyperfrontality in experimentally induced psychosis (ketamine- and psilocybin model of schizophrenia). PMID- 15834760 TI - What is the optimal orthostatic stress to diagnose orthostatic hypotension? PMID- 15834761 TI - Vasovagal fainting as an evolutionary remnant of the fight against hemorrhage. PMID- 15834762 TI - What is the minimum duration of head-up tilt necessary to detect orthostatic hypotension? AB - OBJECTIVE: There is uncertainty as to the minimum duration of head-up tilt (HUT) needed to detect orthostatic hypotension (OH). The orthostatic duration has variably been recommended to be 1, 2, 3, and 5 minutes. The purpose of the current study was 1) to determine the minimum duration of HUT necessary to detect OH and 2) to identify different patterns of orthostatic blood pressure (BP) response in patients with OH. DESIGN/METHODS: We evaluated the medical records of 66 consecutive patients (mean age 70.0+/-10.1 years; 64% male) seen at Mayo Clinic-Rochester from 2000-2001 who fulfilled the criteria for OH (systolic blood pressure [SBP] reduction > or = 20 mmHg within 3 minutes of HUT) during routine clinical autonomic studies. All patients completed an autonomic reflex screen with continuous monitoring of heart rate and BP during supine rest and 5 minutes of 70 degree HUT. Severity of autonomic deficits was quantified with the Composite Autonomic Severity Score (CASS). RESULTS: Overall, BP was the lowest at 1 minute with gradual and partial recovery over the following 4 minutes. Eighty eight percent of patients (N=58) developed OH by 1 minute of HUT, with an additional 11% (N=7) developing OH by 2 minutes and the remaining 1% (N=1) developing OH by 3 minutes. We identified two broad patterns of SBP response to HUT. Forty-eight percent (N=32) of patients demonstrated an initial drop in SBP (> or = 20 mm Hg),which remained stable until tilt-back. Thirty-six percent (N=24) demonstrated an initial drop (> or = 20 mmHg) followed by a progressive decline in SBP until tilt-back. Repeated measures analysis of variance confirmed that the SBP change in response to HUT differed significantly among patients with a stable vs. progressive pattern [F(3,32)=25.1, p<0.001). Patients with the progressive pattern also had more severe adrenergic impairment on the CASS (p=0.03) and were more likely to have their tilt test terminated early (prior to 5 minutes) due to presyncope (p<0.0001) than patients with the stable pattern. CONCLUSIONS: One minute of HUT will detect OH in the great majority (88%) of patients and three minutes will detect the balance. Orthostatic stress beyond 2 minutes is necessary to detect the pattern of progressive OH. Since this group has more severe adrenergic deficits than the group with stable OH, we suggest that the progressive pattern is due to greater impairment of compensatory reflexes. Recognition of the group with progressive fall in BP is important since this group may be at greater risk of orthostatic syncope. PMID- 15834763 TI - Symptoms of autonomic failure in Parkinson's disease: prevalence and impact on daily life. AB - Frequency and clinical importance of autonomic failure in idiopathic Parkinson's disease (PD) are discussed controversially. 141 patients with PD and 50 healthy age-matched control subjects were interviewed for symptoms of autonomic failure and their influence on daily life using a questionnaire. In PD patients, the prevalence of orthostatic dizziness, bladder dysfunction, erectile dysfunction and hyperhidrosis was significantly higher compared with controls. About 50% of PD patients rated the impact of the symptoms of autonomic failure on their daily lives as "a lot" or "very much" due to orthostatic dizziness, bladder dysfunction and constipation, which were more statistically significant than in age-matched controls. Prevalence and number of autonomic symptoms were not correlated with duration and severity of PD. In 32% of patients, impaired cardiovascular regulation was found by standardized cardiovascular function tests. If testing showed abnormal findings, orthostatic dizziness, bladder dysfunction, constipation and erectile dysfunction were significantly more frequent than in patients with normal regulation, but the impact on daily life due to these symptoms differed significantly only for bladder dysfunction between groups. It is concluded that autonomic failure is a clinically relevant, pervasive problem in PD and compromise patients' daily life activities in all stages of the disease. This underlines the necessity to adequately search for and treat these non-dopaminergic symptoms during the whole course of the disease. PMID- 15834764 TI - Cerebral autoregulation is temporarily disturbed in the early recovery phase after dynamic resistance exercise. AB - AIM: To determine cerebral blood-flow velocity (CBFV) and parameters of dynamic cerebral autoregulation (CA) during and after exhausting resistance exercise. METHODS: Strength endurance (23 repetitions) and maximal strength training (8 repetitions) in 16 female and 16 male athletes on a leg curler (m. quadriceps training; approx. 2 s contraction) in the upright position. Registration of ECG, blood pressure by Finapres, CBFV by transcranial Doppler (TCD), and breathing by a Zak breathing-belt. Additional repetitive ergospirometry (O2-uptake, CO2 elimination, ventilation) and blood gas analyses were performed in a subgroup of seven athletes. From BP and CBFV cerebrovascular resistance (CVR), pulsatility index (PI) as well as LF-power, gain and phase-angle (frequency analysis) were derived. RESULTS: All athletes showed significant (p<0.01) 15 % to 30 % increases in CBFV during both training sets without signs of flow depression due to Valsalva maneuvers. In the early recovery, when blood pressure rapidly decreased, CBFV amplitude significantly (p<0.01) increased for 60-80 seconds with mean flow (Vm) at the exercise level, while CVR and PI showed conflicting results, similar to a presyncopal reaction. Ergospirometry and blood gas analyses revealed no evidence of major changes in pCO2, but phase angle was reduced (p<0.001) after exercise, together with an LF power increase (p<0.001). CONCLUSION: An unexpected increase in CBFV amplitude and in Vm occurs directly after dynamic resistance exercise without increased pCO2, which is comparable to a maximum leg press with hypercapnia. CVR and PI results as well as data from frequency analysis show similarities to presyncopal reactions, on the one hand, and point towards a temporarily disturbed cerebral autoregulation, on the other. PMID- 15834765 TI - Cross-spectral analysis of cardiovascular variables in supine diabetic patients. AB - Cardiovascular autonomic neuropathy in diabetes is associated with a high risk of mortality, which makes its early identification clinically important. An easy method for identification of subjects with autonomic dysfunction would be of clinical benefit. We evaluated the autonomic function in 28 diabetic patients and 21 control subjects recording 12 min time series of heart period (RR) and systolic arterial pressure (SAP, Finapres) during supine rest and 60 degrees head up tilt. The power of the high (respiratory) and low (LF approximately 0.1 Hz) frequency oscillations was quantified by spectral analysis. The central frequency of the LF oscillations (LF_freq), phase shift, and the transfer function gain between RR interval and SAP fluctuations were provided by cross-spectral analysis, and measured at the point of maximal coherence. In the supine position 15 patients (LF-) displayed atypical LF variability with the LF_freq being shifted towards lower frequencies (about 0.06 Hz). They also showed larger phase angle, lower values or even absence of coherence and smaller transfer function gain between RR and SAP fluctuations. 13 patients (LF+) and the controls showed the LF_freq around 0.1 Hz, higher coherence and transfer function gain values. The orthostatic maneuver induced the expected changes in the spectral parameters (increase in the LF components of both RR and SAP and decrease in the HF variability of RR) into the LF+ patients and all the control subjects and abnormal response in the other 15 LF-patients. These findings indicate that diabetic subjects with uncharacteristic response to the orthostatic test present abnormal LF variability already in the supine position. Crossspectral parameters while supine may be used for the identification of these subjects. PMID- 15834766 TI - Does peripheral nerve degeneration affect circulatory responses to head-up tilt in spinal cord-injured individuals? AB - Despite the loss of centrally mediated sympathetic vasoconstriction, spinal cord injured (SCI) individuals cope surprisingly well with orthostatic challenges. In the pathophysiology of this intriguing observation spinal sympathetic-, veno arteriolar-(VAR), and myogenic reflexes seem to play a role. The purpose of this study was to assess whether central (stroke volume, heart rate, blood pressure and total peripheral resistance) and peripheral (leg blood flow, leg vascular resistance and femoral arterial diameter) hemodynamic responses to head-up tilt are different in two groups of SCI patients, i. e., SCI individuals with upper motor neuron lesions (who have spinal reflexes, VAR and myogenic reflexes) (U; n=6) and those with lower motor neuron lesion (who have no spinal reflexes, perhaps no VAR due to nerve degeneration, but intact myogenic reflexes) (L; n=5). Ten healthy male individuals served as controls (C) (normal supraspinal sympathetic control and presence of all reflexes). After 10 min supine rest all individuals were tilted to 30 degrees head-up tilt. Red blood cell velocity (measured by echo Doppler ultrasound) in the femoral artery decreased and vascular resistance increased significantly in all three groups in the upright position compared with supine. Mean arterial pressure (MAP) remained unchanged in U and L and increased significantly in C in the upright versus supine position. The present study shows that all SCI individuals were able to maintain MAP by increasing leg vascular resistance during head-up tilt, despite nerve degeneration in L and lack of centrally mediated sympathetic control in all SCI individuals. Results of the present study suggest that not spinal reflexes but local (myogenic) reflex activity plays a pivotal role in peripheral vascular responses upon head-up tilt when central control mechanisms fail. PMID- 15834767 TI - Interval training in elderly men increases both heart rate variability and baroreflex activity. AB - AIMS: Autonomic nervous system activity decreases continuously with age and appears to be a powerful predictor of disease and death. Attempts are thus made to reactivate autonomic drive with the intent of improving health. METHODS: We assessed maximal oxygen consumption (VO2max), autonomic nervous system activity by heart rate variability (HRV) analysis and spontaneous cardiac baroreflex activity (SBR) in eleven elderly men (73.5+/-4.2 years) before and after a 14 week program of intensive cycloergometer interval training. The standard HRV indices were calculated using time domain (mean RR, PNN50, RMSSD, SDNN, SDANN and SDNNIDX), and Fourier transform (total power, ULF,VLF, LF, LFnu, HF, HFnu and LF/HF) analyses of 24-hour, daytime and nighttime Holter recordings. The SBR was calculated from 15-minute recordings of spontaneous blood pressure and RR interval variations using the sequence (slope, slSBR) and cross-spectral (alphaSBRHF and alphaSBRLF) methods. RESULTS: After the training period,VO2max increased by 18.6 % (26.8+/-4.4 to 31.8+/-5.2 ml.kg(-1).min(-1), p<0.01). The nocturnal parasympathetic indices of HRV increased (PNN50: 3.05+/-2.21 to 5.00+/ 2.87%, RMSSD: 29.1+/-7.6 to 38.8+/-10.9 ms, HF: 117+/-54 to 194+/-116 ms2/Hz, all p<0.05) as did the SBR indices (slSBR: 7.0+/-1.8 to 9.8+/-2.1 ms.mmHg(-1), p<0.01; alphaSBRHF: 6.9+/-2.2 to 10.5+/-3.7 ms.mmHg(-1), p<0.05; alphaSBRLF: 5.3+/-2.3 to 6.9+/-3.1 ms.mmHg(-1), p=0.22). CONCLUSION: Intensive endurance training in elderly men enhanced parasympathetic parameters of HRV and, interestingly, of SBR. Physiological mechanisms and long-term clinical effects on health status should be further investigated. PMID- 15834768 TI - The body mass index and level of resection: predictive factors for compensatory sweating after sympathectomy. AB - OBJECTIVE: Compensatory sweating (CS) is the most common adverse event and the main cause of dissatisfaction among patients undergoing a VATS sympathectomy for the treatment of primary hyperhidrosis. It has been described that obese individuals experience more sweating than thinner ones. The aim of this study is to identify the Body Mass Index (BMI) and the level of resection as predictive factors for CS and its relation to levels of patient satisfaction following the procedure. METHODS: From October 1998 to June 2003, 102 patients undergoing VATS sympathectomies (51 for palmar hyperhidrosis, PH, and 51 for axillary hyperhidrosis, AH) were prospectively surveyed. They were divided into three groups according to their BMI: Group I was composed of 19 patients with BMI<20 (9 patients with PH and 10 with AH); Group II was composed of 52 patients with 20 < or =BMI<25 (25 with PH and 27 with AH); and Group III was composed of 31 patients with BMI > or =25 (17 with PH and 14 with AH). Each procedure was simultaneously and bilaterally performed under general anesthesia using two 5.5 mm trocars and a 30 degrees optic system. RESULTS: Patients treated for PH (resection of T2-T3) had more severe CS than those with AH (resection of T3-T4) (p=0.007) and the greater the BMI, the greater the severity of the CS (p<0.001). No statistically significant difference was found between the BMI bands in relation to the degree of satisfaction (p=0.644), nor when we compared the degree of satisfaction to the degree of CS (p=0.316). CONCLUSIONS: The greater the BMI, the more severe the CS, but this did not correlate with the patients' level of satisfaction. Avoiding the resection of T2 sympathetic ganglia is also important in reducing the intensity of CS. PMID- 15834769 TI - Changes in central venous pressure with vasoactive drug injections in humans. AB - We tested whether central venous pressure (CVP) changes during vasoactive drug injections used for baroreflex assessment in humans. We measured CVP during sequential intravenous boluses of nitroprusside (NTP) and phenylephrine (PHE; modified Oxford technique). NTP caused a decrease in CVP of 1.9+/-0.2 mmHg from baseline (P<0.01) and PHE caused an increase in CVP of 0.3+/-0.2 mmHg above baseline. These changes in CVP may contribute to the observed integrated baroreflex responses. PMID- 15834770 TI - Vasovagal syncope and Darwinian fitness. AB - Vasovagal syncope, although often seen as a disease, is the result of a neurophysiological reflex which can be induced in most healthy people after a 30 % reduction in blood volume either by venous pooling or by hemorrhage. Studies in mammals showed that the activation of certain brainstem nuclei including the caudal midline medulla is responsible for hypotension and bradycardia following central hypovolemia. The hypothesis is presented that vasovagal fainting developed during the evolution in order to support hemostasis. Bleeding animals with a central mechanism for the initiation of hypotension had presumably a better chance for blood clot formation and hemostasis than animals with normal blood pressure. In the context of this hypothesis, vasavagal fainting with blood or injury displaying stimuli can be understood as an early attempt to support hemostasis before the development of larger blood losses. PMID- 15834775 TI - Depression prevalence in disadvantaged young black women--African and Caribbean immigrants compared to US-born African Americans. AB - BACKGROUND: Research with Mexican Americans suggests that immigrants have lower rates of mental disorders than U. S.-born Mexican Americans. We examine the prevalence of depression, somatization, alcohol use and drug use among black American women, comparing rates of disorders among U. S.-born, Caribbean-born, and African-born subsamples. METHODS: Women in Women, Infants and Children (WIC) programs, county-run Title X family planning clinics, and low-income pediatric clinics were interviewed using the PRIME-MD. In total, 9,151 black women were interviewed; 7,965 were born in the U. S., 913 were born in Africa, and 273 were born in the Caribbean. RESULTS: Controlling for other predictors, U.S.-born black women had odds of probable depression that were 2.94 times greater than the African-born women (p<0.0001, 95% CI: 2.07, 4.18) and 2.49 times greater than Caribbean-born women (p<0.0016, 95% CI: 1.41, 4.39). Likelihood of somatization did not differ among women who were U. S. born, African born, or Caribbean born. Rates of alcohol and drug problems were exceedingly low among all three groups, with less than 1% of the women reporting either alcohol or drug problems. CONCLUSIONS: These results mirror similar findings for Mexican immigrant as compared with American-born Mexican Americans. The findings suggest that living in the U. S. might increase depression among poor black women receiving services in county entitlement clinics. Further research with ethnically validated instruments is needed to identify protective and risk factors associated with depression in immigrant and U. S.-born poor black women. PMID- 15834776 TI - Acculturation and health status among African-Caribbean, Bangladeshi and White British adolescents--validation and findings from the RELACHS study. AB - OBJECTIVE: This paper reports the validation process for research questions that measure cultural identity in a survey of adolescents from a multi-ethnic urban area. METHOD: Four questions that tapped preferred cultural origins of friends and clothing were used to classify subjects into categories for cultural identity described by Berry (1980). We assessed face, content and construct validity of these questions by: i) investigating the meaning of cultural identity from an African-Caribbean, Bangladeshi, and White British perspective in three focus groups; ii) asking participants to comment on the questions and their suitability for studies of cultural identity; and iii) using data from the Research with East London Adolescents: Community Health Survey (RELACHS) study of adolescents to test hypotheses about associations of acculturation with health outcomes (physical and mental) in order to assess whether these trends are consistent with previous research, and are in accord with trends predicted by the Berry model. RESULTS: Questions about the cultural origins of friends and clothing showed good face and content validity. However, only the questions on friendship choices showed some associations in the predicted directions, with some associations that were unexpected. The most significant findings were that, in comparison with marginalised pupils, for African-Caribbean pupils assimilation was associated with more tiredness (OR=40.7, 3.8-432.8,p<0.001) and traditionalism with less obesity (OR=0.1, 0.02-0.5, p<0.001). For Bangladeshi pupils integration was associated with a lower risk of depression (OR=0.4, 0.2-0.8, p<0.001). CONCLUSION: Questions on friendship choices can be used to assess cultural identity, but associations with health and health risk behaviours are different for each ethnic group. PMID- 15834777 TI - The impact and patterns of hazardous drinking amongst male industrial workers in Goa, India. AB - AIMS: The aim of this study was to describe the impact and patterns of drinking in hazardous drinkers in a male industrial worker population in India. METHOD: A case-control design was used, and 234 subjects (75 hazardous drinkers, 78 casual drinkers and 81 abstinent workers) were randomly selected from a population of male workers who had participated in a survey of drinking behaviour. The definition of these categories was based on the scores of subjects on the Alcohol Use Disorders Identification Test (AUDIT). Subjects were interviewed with a semi structured social and health impact questionnaire, Revised Clinical Interview Schedule (CISR) and Brief Disability Questionnaire (BDQ). A total of 184 spouses of consenting workers were also interviewed with a semi-structured interview, the CISR,BDQ and AUDIT. A sub-group of 55 hazardous drinkers participated in a descriptive study of patterns of drinking, recording drinking behaviour using a daily drink diary over a 1-month period. FINDINGS: Hazardous drinkers have significantly poorer physical and mental health and show trends for adverse social outcomes such as violence. Casual drinkers, on the other hand, were no different from abstinent subjects on any of the key outcomes. As compared to casual drinkers, hazardous drinkers tend to drink alone, in bars, and prefer non commercial alcoholic beverages which are cheaper and have high alcohol concentration. CONCLUSIONS: Hazardous drinking has a significant adverse impact on drinkers and their families. Hazardous drinkers display unique drinking patterns suggesting the role of stigma and preference for higher alcohol containing, but cheaper, drinks available in India. PMID- 15834778 TI - The validity of the Parental Bonding Instrument as a measure of maternal bonding among young Pakistani women. AB - BACKGROUND: Parenting may have an important influence upon women's mental health in societies characterized by strong male gender preference. The Parental Bonding Interview (PBI) has been very widely used, but not yet formally evaluated in a South Asian context. The purpose of this pilot study was to assess the factor structure, face and concurrent validity of the PBI as an assessment of maternal bonding among young Pakistani women. METHODS: A total of 86 women, aged 20-35 years, were identified by door-knocking in two small catchment areas in Islamabad/Rawalpindi, one characterized by higher and one by lower socio-economic status. The PBI, the Clinical Interview Schedule Revised (CIS-R) and the Marital Satisfaction Scale (MSS) were each administered by a female interviewer in the respondent's home. RESULTS: All women agreed to be interviewed privately, the response rate was 100%. Consistent with previous reports, PBI items loaded on two well-characterized internally consistent scales, care and overprotection. In a three-factor solution, the overprotection scale divided into two subscales previously characterized as "Encouragement of Behavioral Freedom" and "Denial of Psychological Autonomy". Concurrent validity was supported by correlations in the expected direction between PBI care and overprotection subscales and both psychological morbidity (CIS-R) and marital satisfaction (MSS). CONCLUSIONS: Pakistani women seem to perceive and respond to PBI items as addressing parental "control" or "overprotection" in a similar similar way to respondents from western Anglophone cultures. This study provides further support for the core construct validity of the PBI, and indicates a perhaps surprising degree of sensitivity to cultural nuances. PMID- 15834780 TI - The use of weights to account for non-response and drop-out. AB - BACKGROUND: Empirical studies in psychiatric research and other fields often show substantially high refusal and drop-out rates. Non-participation and drop-out may introduce a bias whose magnitude depends on how strongly its determinants are related to the respective parameter of interest. METHODS: When most information is missing, the standard approach is to estimate each respondent's probability of participating and assign each respondent a weight that is inversely proportional to this probability. This paper contains a review of the major ideas and principles regarding the computation of statistical weights and the analysis of weighted data. RESULTS: A short software review for weighted data is provided and the use of statistical weights is illustrated through data from the EDSP (Early Developmental Stages of Psychopathology) Study. The results show that disregarding different sampling and response probabilities can have a major impact on estimated odds ratios. CONCLUSIONS: The benefit of using statistical weights in reducing sampling bias should be balanced against increased variances in the weighted parameter estimates. PMID- 15834779 TI - Cross-cultural validity of the structured clinical interview for panic agoraphobic spectrum. AB - BACKGROUND: We sought to develop a series of assessment measures of psychiatric spectrum conditions associated with major DSM-IV mood and anxiety disorders that might capture the true phenotypes underlying these disorders. The specific objective of this report was to describe the methods we employed to create instruments that could cross linguistic and national boundaries and to evaluate the comparability of results obtained when one of these instruments, the Structured Clinical Interview for Panic-Agoraphobic Spectrum (SCI-PAS), was administered in the United States and in Italy. METHOD: After developing, in parallel, the English and the Italian versions of the SCI-PAS, identical protocols were conducted in patients and control samples at the University of Pittsburgh and the University of Pisa to examine the reliability and validity of the interview. RESULTS: Total and domain scores on the SCI-PAS were strikingly similar in the US and Italian patient groups and in controls. In addition, similarly high levels of inter-rater and test-retest reliability were found at the two sites. Finally, virtually identical patterns of relationships were found between the domains of the SCI-PAS and established measures of the same constructs. CONCLUSIONS: The SCI-PAS displays similar reliability and validity properties in the two versions. This suggests that the instrument taps a phenotype that is consistent in American and Italian patient and control populations. PMID- 15834781 TI - Dual work and family roles and depressive symptoms in two birth cohorts of women. AB - BACKGROUND: Rising depression rates in more recent cohorts of women have been attributed in part to their increased participation in dual family and work roles. METHOD: This study examined associations among depressive symptoms, age, and work and marital status in two cohorts of women, all mothers, born between 1931 and 1944 (preboomers) or between 1945 and 1958 (baby boomers), assessed at comparable ages. RESULTS: Being married (vs. divorced) was related to less depression within and across cohorts, whereas working was related to more depression in preboomers only. Moreover, divorced working preboomers were significantly more depressed than women in most other role status groups within and across cohorts. Depression scores declined across age among working women in the combined cohorts; however, that association held only for baby boomers when cohorts were analyzed separately. Among divorced working women, that decline was significantly greater in baby boomers than preboomers. CONCLUSIONS: These cohort differences support a call for new social policies that address the mental health needs of women and their children. PMID- 15834783 TI - A comparative study of diagnostic practice in psychiatry in Northern Norway and Northwest Russia. AB - BACKGROUND: The co-operation between psychiatrists in Norway and Russia is increasing. The object of this study was to find out whether there were differences in diagnostic practice of psychiatrists in both countries, to look at the nature of the differences and to examine whether these differences affected diagnostic quality. METHOD: Thirty medical doctors working at psychiatric hospitals in both countries diagnosed 12 clinical case vignettes selected from a wide spectre of psychiatric disorders. RESULTS: The Russian clinicians used a larger range of diagnoses than the Norwegians. The Russians tended to diagnose schizophrenia and schizophrenia-like disorders in cases that presented psychotic syndromes, and somatoform disorders in cases that presented agoraphobia. The Norwegians tended to evaluate affective aspects in preference to psychotic symptoms in the case of schizoaffective disorder and overestimate the degree of depression. In general, the Russians had lower total score of correct answers than the Norwegians. CONCLUSION: In spite of the limitations due to minor differences in the data collection phase in the two countries, the study clearly demonstrates differences in diagnostic practice between the countries. PMID- 15834782 TI - Mental disorders associated with benzodiazepine use among older primary care attenders--a regional survey. AB - BACKGROUND: Benzodiazepines (BDZs) are among the most commonly prescribed drugs in the elderly and their use is often too prolonged according to current international guidelines. METHODS: We investigated the pattern of use of BDZs among 65- to 84-year-old attenders at 40 Italian primary care practices who answered positively in a questionnaire on the use of anxiolytic/hypnotic drugs and successfully completed the PRIME-MD questionnaire. The survey lasted 6 months, from February to July 2001. RESULTS: Of the 1,156 subjects who completed the PRIME-MD, 748 subjects were positive for at least one psychiatric diagnosis. A depressive disorder was present in 36.5% of all PRIME-MD completers and in about one-third of cases represented by major depression, whereas anxiety disorders, somatoform disorders and alcohol abuse accounted for 18.2%, 8.1% and 1.7% of all PRIME-MD completers, respectively. Most patients were first prescribed BDZs by their General Practitioners (GPs) for each diagnostic group. However, patients with pure anxiety started BDZ treatment during hospitalisation more often (19.7%) than patients with depressive disorders (13.7 %). Moreover, patients with comorbid anxiety and depressive disorders (CADD) were most likely to receive their first BDZ prescription from a psychiatrist (15.7 %). Sleep disturbances were present in at least 50% of cases (and up to 86 %) in each diagnostic group. About 75% of prescriptions concerned anxiolytic BDZ or medium/long-acting BDZ. Most patients with anxiety, CADD and depression used night-time BDZ (65.2%, 56.9 % and 60.5%, respectively). The prevalence of chronic use of BDZs was equally high in all categories of psychiatric disorder (about 90 % for each), showing that the vast majority of patients, irrespective of the diagnosis, had been using BDZs for years. CONCLUSIONS: BDZs are widely prescribed for elderly people by their GPs, often for a considerable length of time. The evidence that many BDZ consumers suffer from a depressive or an anxiety disorder, or both, could be a starting point for encouraging a rational prescription in accordance with international guidelines. PMID- 15834784 TI - Gunshot suicides in England--a multicentre study based on coroners' records. AB - BACKGROUND: Gunshot suicides account for 2.5% of suicides in England and Wales. This amounts to more than 100 deaths per year. Information about such deaths may assist in the development of suicide prevention strategies. METHOD: We have examined coroners' inquest records for all gunshot suicides between 1st January 2000 and 31st December 2001 in 24 coroners' jurisdictions in England. RESULTS: Fifty-eight gunshot suicides were identified, including one homicide-suicide. Ninety-three per cent of cases were male. Sport or occupational usage was the main reason for owning the gun. Ten per cent were farmers or farm-workers. In 20% of cases the gun did not belong to the individual who used it for suicide. This was more likely in younger suicides. Seven (12.1%) individuals used illegally owned handguns. Large amounts of alcohol had been consumed before the act in nine cases. Nearly three-quarters (72.9%) of individuals with diagnostic information had a probable diagnosis of depression. However, only 22.4% had ever had contact with psychiatric services. Two shotgun certificate holders were under the care of psychiatric services at the time of their death and two others had a history of previous self-harm. CONCLUSIONS: Strategies to reduce the number of gunshot suicides need to focus on limiting access to guns. These include restricting access to guns by non-certificate holders and those who may be at increased risk of suicide, and holding regular gun amnesties. PMID- 15834785 TI - Caregiver-coping in bipolar disorder and schizophrenia--a re-examination. AB - BACKGROUND: The caregiving experience has been extensively investigated in some chronic/severe mental illnesses such as schizophrenia. These studies have suggested that illness variables and situational/personal characteristics of caregivers have a significant influence on how caregivers cope with mental illness. However, other similar conditions, e. g. bipolar affective disorder (BPAD), have been relatively neglected in this regard. This study attempted to compare caregiver-coping in BPAD and schizophrenia and to explore the determinants of such coping. METHOD: Illness variables and coping, burden, appraisal, perceived support, and neuroticism among caregivers were examined in 50 patients each of BPAD and schizophrenia and their caregivers. RESULTS: High levels of patient-dysfunction and caregiver-burden, low awareness of illness and low perceived control over patient's behaviour were characteristic of both BPAD and schizophrenia, with no significant differences between the two groups on these parameters. Coping patterns were also quite alike, though caregivers of patients with schizophrenia were using some emotion-focused strategies significantly more often. Caregiver's gender, patient-dysfunction and caregiver neuroticism had a significant influence on coping patterns, but explained only a small proportion of the variance in use of different coping strategies. CONCLUSIONS: Coping and other elements of the caregiving experience in BPAD are no different from schizophrenia. The relationship between caregiver-coping and its determinants appears to be a complex one. More methodologically sound and culturally relevant investigations are required to understand this intricate area, with the hope that a better understanding will help the cause of both patients and their caregivers. PMID- 15834786 TI - Hans Gerhard Creutzfeldt (1885-1964): a life in neuropathology. PMID- 15834787 TI - Point mutation of (+)-germacrene A synthase from Ixeris dentata. AB - Sesquiterpene cyclases catalyze the conversion of common precursor, farnesyl pyrophosphate, into various terpene backbones. X-ray crystallography of tobacco epi-aristolochene synthase has previously proposed a cyclization mechanism wherein the allylic carbocation intermediate is stabilized by the main chain carbonyl oxygens of three consecutive threonine residues. Alignment of amino acid sequences of plant terpene cyclases shows that the first position of the triad is almost invariably threonine or serine. To probe the carbocation-stabilizing role, the amino acid residues of the 433TSA435 triad in (+)-germacrene A synthase from Ixeris dentata were altered by site-directed mutagenesis. Enzyme kinetic measurements of the mutants and GC/MS analysis of the enzyme reaction products indicate that mutations of the triad decreased enzyme catalysis rather than substrate binding but did not affect its structural rearrangement in the catalytic mechanism. This is the first report that the hydroxyl group of threonine at the first position of the triad is required for the cyclase activity. PMID- 15834788 TI - Development and characterization of a recombinant chicken single-chain Fv antibody detecting Eimeria acervulina sporozoite antigen. AB - Chicken monoclonal antibody (mAb), 8C3, which is reactive with a sporozoite antigen of Eimeria acervulina, is a potential therapeutic agent against avian coccidiosis caused by Eimeria spp. However, production of large amounts of 8C3 mAb in cell culture system is labor intensive and not cost-effective. Accordingly, recombinant single chain variable fragment (ScFv) antibody was constructed by amplification of the V(H) and V(L) genes from chicken hybridoma, 8C3 and when expressed in E. coli gave 5 mg l(-1). The expressed protein showed antigen binding activity equivalent to that of the parental mAb. In addition, nucleotide sequence comparison of 8C3 gene to the germline chicken V(L) genes suggested that the gene conversion with (V)lambda pseudogenes might contribute to the diversification of V(L) genes in chickens. PMID- 15834789 TI - Enhanced radical scavenging activity of genetically modified Arabidopsis seeds. AB - The proanthocyanidin (PA) content was increased in seeds of pap1-D mutant of Arabidopsis thaliana, in which the expression of endogenous PAP1 gene encoding a Myb-like transcription factor was induced by activation-tagging with enhancer sequences derived from cauliflower mosaic virus 35S promoter. In contrast, the PA contents decreased in seeds of transgenic plants transformed with a PAP1 cDNA or with a PAP1 chimeric repressor, although the amount of soluble anthocyanins increased in seeds of transgenic plants over-expressing PAP1 cDNA. The enhanced radical scavenging activity was observed only in the seed extracts of pap1-D mutant, indicating that PAs are primarily responsible for radical scavenging activity in seeds. The present study suggests the feasibility of engineering a transcription factor of flavonoid biosynthesis for health beneficial plant seeds. PMID- 15834790 TI - Gluconobacter oxydans NAD-dependent, D-fructose reducing, polyol dehydrogenases activity: screening, medium optimisation and application for enzymatic polyol production. AB - Gluconobacter oxydans LMG 1489 was selected as the best strain for NAD(P) dependent polyol dehydrogenase production. The highest enzyme activities were obtained when this strain was cultivated on a medium consisting of 30 g glycerol l(-1), 7.2 g peptone l(-1) and 1.8 g yeast extract l(-1). Two D-fructose reducing, NAD-dependent intracellular enzymes were present in the G. oxydans cell free extract: sorbitol dehydrogenase, and mannitol dehydrogenase. Substrate reduction occurred optimally at a low pH (pH 6), while the optimum for substrate oxidation was situated at alkaline pHs (pH 9.5-10.5). The mannitol dehydrogenase was more thermostable than the sorbitol dehydrogenase. The cell-free extract could be used to produce D-mannitol and D-sorbitol enzymatically from D-fructose. Efficient coenzyme regeneration was accomplished by formate dehydrogenase mediated oxidation of formate into CO2. PMID- 15834791 TI - Purification and characterization of an alkaline cellulase from a newly isolated alkalophilic Bacillus sp. HSH-810. AB - An alkaline cellulase from Bacillus sp. HSH-810 was purified 8.7-fold with a 30% yield and a specific activity of 71 U mg(-1) protein. It was optimally active at pH 10 and 50 degrees C and was stable from pH 6 to 10 with more than 60% activity remaining after heating at 60 degrees C for 60 min. The molecular mass of cellulase was 80 kDa. It was inhibited by 50% by Fe3+ (1 mM) and Mn2+ (0.1 mM) but was relatively insensitive to Hg2+ and Pb2+ at 1 mM. PMID- 15834792 TI - In vitro study of alginate-chitosan microcapsules: an alternative to liver cell transplants for the treatment of liver failure. AB - The application of alginate-chitosan (AC) microcapsules to liver cell transplantation has not been previously investigated. In the current in vitro study, we have investigated the potential of AC microcapsules for the encapsulation of liver cells and show that the AC membrane supports the survival, proliferation and protein secretion by entrapped hepatocytes. The AC membrane provides cell immuno-isolation and has the potential for cell cryopreservation. The AC microcapsule has several advantages compared to more widely used alginate poly-L-lysine (APA) microcapsules for the application of cell therapy. PMID- 15834793 TI - Textile dye decolorization using cyanobacteria. AB - Cyanobacterial cultures isolated from sites polluted by industrial textile effluents were screened for their ability to decolorize cyclic azo dyes. Gloeocapsa pleurocapsoides and Phormidium ceylanicum decolorized Acid Red 97 and FF Sky Blue dyes by more than 80% after 26 days. Chroococcus minutus was the only culture which decolorized Amido Black 10B (55%). Chlorophyll a synthesis in all cultures was strongly inhibited by the dyes. Visible spectroscopy and TLC confirmed that color removal was due to degradation of the dyes. PMID- 15834794 TI - Expression of Escherichia coli AppA2 phytase in four yeast systems. AB - To develop an effective fermentation system for producing Escherichia coliphytase AppA2, we expressed the enzyme in three inducible yeast systems: Saccharomyces cerevisiae (pYES2), Schizosaccharomyces pombe (pDS472a), and Pichia pastoris (pPICZ alphaA), and one constitutive system: P. pastoris (pGAPZalphaA). All four systems produced an extracellular functional AppA2 phytase with apparent molecular masses ranging from 51.5 to 56 kDa. During 8-day batch fermentation in shaking flasks, the inducible Pichia system produced the highest activity (272 units ml(-1) medium), whereas the Schizo. pombe system produced the lowest activity (2.8 units ml(-1)). The AppA2 phytase expressed in Schizo. pombe had 60 75% lower K(m)for sodium phytate and 28% higher heat-stability at 65 degrees C than that expressed in other three systems. However, all four recombinant AppA2 phytases had pH optimum at 3.5 and temperature optimum at 55 degrees C and similar efficacy in hydrolyzing phytate-phosphate from soybean meal. PMID- 15834795 TI - Immobilization of beta-fructofuranosidase from Aspergillus japonicus on chitosan using tris(hydroxymethyl)phosphine or glutaraldehyde as a coupling agent. AB - A partially purified beta-fructofuranosidase from Aspergillus japonicus was covalently immobilized on to chitosan beads using either glutaraldehyde or tris(hydroxymethyl)phosphine (THP) as a coupling agent. Compared with the glutaraldehyde-immobilized and the free enzyme, the THP-immobilized enzyme had the highest thermal stability with 78% activity retained after 12 days at 37 degrees C. The THP-immobilized enzyme also had higher reusability than that immobilized by glutaraldehyde, 75% activity was retained after 11 batches (or 11 days) at 37 degrees C for the THP immobilized enzyme system. Less yield (48%) of fructooligosaccharides (FOS) were produced by the THP-immobilized enzyme compared with the free enzyme system (58%) from 50 (w/v) sucrose at 50 degrees C. PMID- 15834796 TI - Multiple gene copy number enhances insulin precursor secretion in the yeast Pichia pastoris. AB - We have found a direct relationship between protein production in Pichia pastoris and the number of introduced synthetic genes of miniproinsulin (MPI), fused to the Saccharomyces cerevisiae pre-pro alpha factor used as secretion signal, and inserted between the alcohol oxidase 1 (AOX1) promoter and terminator sequences. Two consecutive approaches were followed to increase the number of integrated cassettes: the head-to-tail expression cassette multimerization procedure and re transformation with a dominant selection marker. This increased expression from 19 to 250 mg l(-1) when about 11 copies have been integrated. Further, the correct position of one of the disulphide bridges of the purified molecule was verified by digestion with Glu-C endoprotease, followed by mass spectrometry of the isolated fragments. PMID- 15834797 TI - Cell volume distribution dynamics of Chlorella vulgaris Beij. in batch cultures under continuous light. AB - Cell volume distribution in Chlorella vulgaris cultures coming out of senescence was measured by flow cytometry every 6 h for 114 h in a full-factorial experiment with initial nitrate (420-4200 microg NO3-N l(-1)), phosphate (9-186 microg PO4-P l(-1)), and continuous light (50-330 microE m(-2) s(-1)) as treatments. The maxima in median and median absolute deviation (MAD) of cell volume were achieved within 6 h of each other and their timing was not affected by any treatment. Population specific growth rate during the first 66 h calculated from volume distribution changes was significantly affected by light treatment only (p=0.002). PMID- 15834798 TI - Domain engineering of Saccharomyces cerevisiae exoglucanases. AB - To illustrate the effect of a cellulose-binding domain (CBD) on the enzymatic characteristics of non-cellulolytic exoglucanases, 10 different recombinant enzymes were constructed combining the Saccharomyces cerevisiae exoglucanases, EXG1 and SSG1, with the CBD2 from the Trichoderma reesei cellobiohydrolase, CBH2, and a linker peptide. The enzymatic activity of the recombinant enzymes increased with the CBD copy number. The recombinant enzymes, CBD2-CBD2-L-EXG1 and CBD2-CBD2 SSG1, exhibited the highest cellobiohydrolase activity (17.5 and 16.3 U mg(-1) respectively) on Avicel cellulose, which is approximately 1.5- to 2-fold higher than the native enzymes. The molecular organisation of CBD in these recombinant enzymes enhanced substrate affinity, molecular flexibility and synergistic activity, contributing to their elevated action on the recalcitrant substrates as characterised by adsorption, kinetics, thermostability and scanning electron microscopic analysis. PMID- 15834800 TI - Flavonoids: from cell cycle regulation to biotechnology. AB - Flavonoids have been proposed to play diverse roles in plant growth and development, including defense, symbiosis, pollen development and male fertility, polar auxin transport, and protection against ultraviolet radiation. Recently, a new role in cell cycle regulation has emerged. Genetic alteration of glucuronide metabolism by altered expression of a Pisum sativum UDP-glucuronosyltransferase (PsUGT1) results in an altered cell cycle in pea, alfalfa, and Arabidopsis. In alfalfa, altered expression of PsUGT1 results in accumulation of a flavonoid-like compound that suppresses growth of cultured cells. The results are consistent with the hypothesis that PsUGT1 functions by controlling cellular levels of a factor controlling cell cycle (FCC). PMID- 15834801 TI - Development of a candidate vaccine for Newcastle disease virus by epitope display in the Cucumber mosaic virus capsid protein. AB - A peptide fusion to the capsid protein (CP) of Cucumber mosaic virus(CMV) was designed to express either a 17 amino acid (aa) neutralizing epitope of the Newcastle disease virus (NDV) fusion (F) protein or an eight aa neutralizing epitope of the NDV hemagglutinin-neuraminidase (HN) protein. Fusions of the F, HN and HN2 (duplicated HN epitope) were made in the internal betaH-betaI loop (motif 5) within the CMV CP. Recombinant RNA3 transcripts of the Ixora strain of CMV were inoculated on to Nicotiana benthamiana, together with CMV RNA1 and CMV RNA2. When the F and HN epitopes were placed in the internal motif, the modified virus was infectious and the HN NDV epitope was recognized by anti-NDV sera. However, in some plants, deletions of one to several of the inserted amino acids occurred. A duplication of the HN epitope rendered the virus non-viable. PMID- 15834802 TI - Enhancement of enzyme activity and enantioselectivity by cyclopentyl methyl ether in the transesterification catalyzed by Pseudomonas cepacia lipase co-lyophilized with cyclodextrins. AB - The solvent effects of cyclopentyl methyl ether (CPME) on the reaction rates and enzyme enantioselectivity in the enantioselective transesterifications of racemic 6-methyl-5-hepten-2-ol (racemic sulcatol: SUL) and racemic 2,2-dimethyl-1,3 dioxolane-4-methanol (racemic solketal: SOL) with a series of enol esters catalyzed by Pseudomonas cepacia lipase co-lyophilized with cyclodextrins (alpha , beta-, gamma-, partially methylated beta-, and 2,3,6-tri-O-methyl-beta cyclodextrin: alphaCyD; betaCyD; gammaCyD; Me1.78betaCyD; Me3betaCyD) were investigated and compared with those in diisopropyl ether (IPE). In the case of SUL, enzyme activities of the co-lyophilizate with Me1.78betaCyD in CPME were lower than those in IPE with every acyl source, however, the absolute enantiopreference was shown in the transesterification with vinyl butyrate (VBR) in IPME. When the substrates were SOL and VBR, the enzyme activities in CPME were greatly enhanced as high as 1.6-9.8-fold, while the enantioselectivities in CPME were comparable to those in IPE. PMID- 15834803 TI - Enhancement of polyunsaturated fatty acid production by cerulenin treatment in polyunsaturated fatty acid-producing bacteria. AB - When docosahexaenoic acid (DHA)-producing Moritella marina strain MP-1 was cultured in the medium containing 0.5 microg cerulenin ml-1, an inhibitor for fatty acid biosynthesis, the cells grew normally, but the content of DHA in the total fatty acids increased from 5.9-19.4%. The DHA yield of M. marina strain MP 1 cells also increased from 4 to 13.7 mg l-1 by cerulenin treatment. The same effect of cerulenin was observed in eicosapentaenoic acid (EPA)-producing Shewanella marinintestina strain IK-1 grown in the medium containing 7.5 microg cerulenin ml-1, and the cerulenin treatment increased the EPA yield from 1.6 to 8 mg l-1. The use of cerulenin is, therefore, advantageous to increase the content of intracellular polyunsaturated fatty acids (PUFA) in particular PUFA-containing phospholipids in bacterial cells. PMID- 15834804 TI - Growth and metabolism of marine fish Chinook salmon embryo cells: response to lack of glucose and glutamine. AB - A peculiar phenomenon, differing from the response of mammalian cells, occurred when Chinook salmon embryo (CHSE) cells were passaged in the medium lacking of both glucose and glutamine. To elucidate metabolic mechanism of CHSE cells, the metabolism parameters, key metabolic enzymes, and ATP levels were measured at different glucose and glutamine concentrations. In the glutamine-free culture, hexokinase activity kept constant, and lactate dehydrogenase (LDH) activity decreased. This indicated that lack of glutamine did not expedite glucose consumption but made it shift to lower lactate production and more efficient energy metabolism. The results coincided with the experimental results of unaltered specific glucose consumption rate and decreased yield coefficients of lactate to glucose. In the glucose-free culture, simultaneous increase of glutaminase activity and of specific ammonia production rate suggested an increased flux into the glutaminolysis pathway, and increases of both glutamate dehydrogenase activity and yield coefficient of ammonia to glutamine showed an increased flux into deamination pathway. However, when glucose and glutamine were both lacking, the specific consumption rates of most of amino acids increased markedly, together with decrease of LDH activity, indicating that pyruvate derived from amino acids, away from lactate production, remedied energy deficiency. When both glucose and glutamine were absent, intracellular ATP contents and the energy charge remained virtually unaltered. PMID- 15834805 TI - Production of active pigment epithelium-derived factor inE. coli. AB - Human pigment epithelium-derived factor (PEDF), a neurotrophic factor, is the most potent natural inhibitor of angiogenesis. To produce the active PEDF, the gene coding for the human PEDF protein was expressed in E. coli. The rPEDF protein was expressed at 457 mg l-1 as a soluble protein. The yield of purified GST fusion protein was 14 mg l-1. Purified rPEDF inhibited tube formation in endothelial cells. PMID- 15834806 TI - Neural network designs for poly-beta-hydroxybutyrate production optimization under simulated industrial conditions. AB - Improvement of the fermentation efficiency of poly-beta-hydroxybutyrate (PHB) may make it competitive with chemically synthesized petroleum-based polymers. One step toward this is optimization of fluid dispersion and the feed rates to a fed batch bioreactor. In a recent study using a fermentation model, dispersion corresponding to a Peclet number of approximately 20 was shown to maximize the productivity of PHB. Here further improvement has been investigated using neural optimization. A comparison of seven neural topologies has shown that while feed forward and radial basis neural networks are computationally efficient, recurrent networks generate higher concentrations of PHB. All networks enhanced the productivity by 16-93% over model-based optimization. PMID- 15834807 TI - Surface properties of Yarrowia lipolytica and their relevance to gamma decalactone formation from methyl ricinoleate. AB - The surface of the lipid-degrading yeast, Yarrowia lipolytica, was characterized by contact angle and zeta potential (zeta) measurements. The cells were mainly hydrophilic with a negative charge that was only affected from pH 2 to 4. To study the effects of the surface charges on the biotransformation of methyl ricinoleate into the aroma compound, gamma-decalactone, the zeta values of the substrate droplets were modified by adding a cationic surfactant into the medium at concentrations that did not diminish cell viability: the adhesion of the lipid substrate to the cells was increased but not the overall performance of the process, therefore the adhesion is not the rate limiting here. Our methodology offers interesting perspectives for further applications. PMID- 15834809 TI - Variation of xylanosomal subunit composition of Streptomyces olivaceoviridis by nitrogen sources. AB - Besides affecting the xylanases production, different nitrogen sources present in the media also caused changes in the xylanosomal subunit composition of Streptomyces olivaceoviridis E-86. Four xylanosome fractions, purified from the culture supernatant of S. olivaceoviridis E-86 grown on different nitrogen sources, exhibited high specificity towards different xylans and were composed of different subunits. Thus, S. olivaceoviridis E-86 regulates the expression of xylanase activity and varies the xylanosome composition according to the nitrogen sources possibly through the action of the secreted proteases. PMID- 15834808 TI - Cloning and DNA-binding properties of ethylene response factor, LeERF1 and LeERF2, in tomato. AB - Two new genes, LeERF1 andLeERF2, were isolated from a tomato (Lycopersicon esculentum cv. Lichun) cDNA library. Phylogenetic analysis indicated that they encoded Ethylene Responsive Element Binding Proteins (EREBPs), characterized by a conserved ERF (ethylene response factor) domain of specific binding plant cis acting elements GCC box. Both LeERF1 and LeERF2 proteins were obtained via prokaryotic expression and purification. Electrophoretic mobility shift assay showed that LeERF1 and LeERF2 protein could bind to the promoter of the NP24 gene coding for pathogenesis-related protein osmotin precursor but not the mutant promoter where its GCC box was deleted. Polyclonal antibodies of LeERF1 and LeERF2 blocked their binding in vitro. PMID- 15834810 TI - Expression of a self-processing, pathogen resistance-enhancing gene construct in Arabidopsis. AB - A gene cassette, p35S-CNO, was designed to express three gene products driven by a single constitutive CaMV 35S promoter. The individual coding regions were linked in frame to produce a single polyprotein, using spacer sequences encoding a specific heptapeptide cleavage recognition site (ENLYFQS) for the nuclear inclusion-a (NIa) proteinase of tobacco etch virus (TEV). The protein coding sequences used were: a Trichoderma harzinum endochitinase, a truncated NIa proteinase of TEV, and a wheat oxalate oxidase. When p35S-CNO construct was tested in Arabidopsis thaliana, the polyprotein was properly cleaved after translation and the products exhibited functional enzymatic activity in vivo. PMID- 15834811 TI - Perinuclear translocation of hsp 27 in shear stress exposed human endothelial cells. AB - Expression of hsp 27 in human umbilical vein endothelial cells exposed to a shear stress was investigated. Using immunostaining, it was concluded that shear stress results in perinuclear translocation of hsp 27. Polymerization of actin microfilaments plays an important role in this process. PMID- 15834813 TI - Rational inferences about departures from Hardy-Weinberg equilibrium. AB - Previous studies have explored the use of departure from Hardy-Weinberg equilibrium (DHW) for fine mapping Mendelian disorders and for general fine mapping. Other studies have used Hardy-Weinberg tests for genotyping quality control. To enable investigators to make rational decisions about whether DHW is due to genotyping error or to underlying biology, we developed an analytic framework and software to determine the parameter values for which DHW might be expected for common diseases. We show analytically that, for a general disease model, the difference between population and Hardy-Weinberg expected genotypic frequencies (delta) at the susceptibility locus is a function of the susceptibility-allele frequency (q), heterozygote relative risk (beta), and homozygote relative risk (gamma). For unaffected control samples, is a function of risk in nonsusceptible homozygotes (alpha), the population prevalence of disease (KP), q, beta, and gamma. We used these analytic functions to calculate and the number of cases or controls needed to detect DHW for a range of genetic models consistent with common diseases (1.1 < or = gamma < or = 10 and 0.005 < or = KP < or = 0.2). Results suggest that significant DHW can be expected in relatively small samples of patients over a range of genetic models. We also propose a goodness-of-fit test to aid investigators in determining whether a DHW observed in the context of a case-control study is consistent with a genetic disease model. We illustrate how the analytic framework and software can be used to help investigators interpret DHW in the context of association studies of common diseases. PMID- 15834814 TI - The scholarship of teaching in health science schools. AB - Teaching is a core mission for all health science schools. Despite its key role in training new generations of health care professionals, teaching has been overshadowed by the revenue- and prestige-generating activities of research and clinical care. Research, both basic and clinical, is equated with scholarship and is rewarded in the promotion and tenure process, as well as with intramural and extramural funding. Clinical service generates revenue for schools; teaching, however, does not generate revenue, and, traditionally, teaching and the creative activity related to it have been seen not as scholarship but as an expectation. Over the last decade or so, scholars of teaching have called for a new view of scholarship that includes the scholarship of teaching. This view is broader in scope than scholarly teaching within a classroom or clinic. It refers to scholarly activity related to teaching that results in enduring products that are peer reviewed and broadly disseminated. These are examples of scholarly work and should be recognized as such. Academic institutions should value high-quality teaching and educational innovations and reward them as scholarly work. This article presents an overview of what the scholarship of teaching means, how it can be assessed, and the needed next steps. PMID- 15834815 TI - Mentors, colleagues, and successful health science faculty: lessons from the field. PMID- 15834816 TI - Problem Based Learning: an introduction and overview of the key features of the approach. AB - Problem Based Learning (PBL) has been adopted in educational programs in a variety of disciplines, including veterinary medicine. There is a voluminous literature on the subject, but it often remains unclear just what is being done in the name of PBL, and different accounts highlight different, often contradictory, positions on the key features of the approach. Similarly, despite the many claims made for the advantages of PBL, the evidentiary basis of such claims is often questionable. This article provides an introductory overview of what appear to be the key features of the approach and a brief summary of empirical evidence on its effectiveness. PMID- 15834817 TI - Progress in the capture, manipulation, and delivery of medical media and its impact on education, clinical care, and research. AB - In this article, advances in the application of medical media to education, clinical care, and research are explored and illustrated with examples, and their future potential is discussed. Impact is framed in terms of the Sloan Consortium's five pillars of quality education: access; student and faculty satisfaction; learning effectiveness; and cost effectiveness. (Hiltz SR, Zhang Y, Turoff M. Studies of effectiveness of learning networks. In Bourne J, Moore J, ed. Elements of Quality Online Education. Needham, MA: Sloan-Consortium, 2002:15 45). The alternatives for converting analog media (text, photos, graphics, sound, video, animations, radiographs) to digital media and direct digital capture are covered, as are options for storing, manipulating, retrieving, and sharing digital collections. Diagnostic imaging is given particular attention, clarifying the difference between computerized radiography and digital radiography and explaining the accepted standard (DICOM) and the advantages of Web PACS. Some novel research applications of medical media are presented. PMID- 15834818 TI - Understanding the interrelationship of instructional technology use and organizational culture: a case study of a veterinary medical college. AB - Many predicted that in the latter part of the twentieth century modern technology would revolutionize higher education and "create a second Renaissance" (Sculley J. The relationship between business and higher education: A perspective on the 21st century. Commun ACM32:1056-1061, 1989 p1061). However, as the reality of the twenty-first century has set in, it is apparent that these revolutionary prophecies have fallen short. Using the lens of Douglas's Typology of Grid and Group, this case study examines (1) the organizational context of a veterinary medical college at a large Midwestern university; (2) individual faculty members' preferences toward instructional technology use; and (3) the interrelationship of culture and the decision process to implement instructional technology use in curricula. The study has several implications for instructional technology use in veterinary medical educational settings that help explain how cultural context can guide leadership decisions as well as influence faculty motivation and preference. The findings suggest that a key mitigating factor to instructional technology implementation is conflict or concord between the cultural biases of faculty members and actual cultural identity of the college (Stansberry S, Harris EL. Understanding why faculty use (or don't use) IT: Implementation of instructional technology from an organizational culture perspective. In Simonson M, Crawford M, eds. 25th Annual Proceedings: Selected Research and Development Papers Presented at the 2002 National Convention of the Association for Educational Communications and Technology, vol. 1. North Miami Beach, FL: Nova Southeastern University:viii, 507). PMID- 15834819 TI - The Edinburgh Electronic Veterinary Curriculum: an online program-wide learning and support environment for veterinary education. AB - The Edinburgh Electronic Veterinary Curriculum (EEVeC) is a purpose-built virtual learning support environment for the veterinary medicine program at the University of Edinburgh. It is Web based and adapted from a system developed for the human medical curriculum. It is built around a set of databases and learning objects and incorporates features such as course materials, personalized timetables, staff and student contact pages, a notice board, and discussion forums. The EEVeC also contains global or generic resources such as information on quality enhancement and research options. Many of these features contribute to the aim of building a learning community, but a challenge has been to introduce specific features that enhance student learning. One of these is a searchable lecture database in which learning activities such as quizzes and computer-aided learning exercises (CALs) can be embedded to supplement a synopsis of the lecture and address the key needs of integration and reinforcement of learning. Statistics of use indicate extensive student activity during evenings and weekends, with a pattern of increased usage over the years as more features become available and staff and students progressively engage with the system. An essential feature of EEVeC is its flexibility and the way in which it is evolving to meet the changing needs of the teaching program. PMID- 15834820 TI - Ten years of CLIVE (Computer-Aided Learning in Veterinary Education) in the United Kingdom. AB - This paper outlines the work of the CLIVE (Computer-Aided Learning in Veterinary Education) project over a 10-year period, set against the backdrop of changes in education policy and learning technology developments. The consortium of six UK veterinary schools and 14 international Associate Member Schools has been very successful. Sustaining these partnerships requires that the project redefine itself and adapt to cater to the diverse learning needs of today's students and to changing professional and societal needs on an international scale. PMID- 15834821 TI - "Blended" online technology: maximizing instructor reach. AB - This article describes the development and implementation of a small animal nutrition course for veterinary technicians using ''blended'' online technology and delivery at multiple sites. Student contact was accomplished through both synchronous and asynchronous Internet platforms. A summary of student evaluations is provided. Application of the process described here may be appropriate where specific content expertise is available in one location and desired in multiple locations. Unique features of the course include (a) course and instructor provided by industry partner, (b) synchronous and asynchronous blended online elements, and (c) multiple schools engaged in merged, simultaneous sessions. PMID- 15834822 TI - Teaching the didactic aspects of ophthalmology and dermatology using an off-site instructor. AB - A shortage of veterinary educators in various clinical disciplines is requiring faculty at veterinary schools to develop novel approaches to teaching professional students a core of information in each major clinical discipline. This article presents experiences at Washington State University (WSU) College of Veterinary Medicine in the teaching of the didactic components of ophthalmology and dermatology over four years using pre-prepared lectures developed by off-site instructors. The methodology of lecture preparation and presentation and various perspectives of student satisfaction with this teaching/learning method are presented. PMID- 15834823 TI - Surveys concerning the needs for and evaluation of distance learning in veterinary epidemiology. AB - Distance learning courses in veterinary epidemiology, taught in French and spread over half a year, were organized in 2003 and 2004. They were the subject of surveys concerning need, evaluation, attaining learning objectives, and satisfaction, the results of which will be used to improve the course scheduled to take place in 2005. PMID- 15834824 TI - Using 3D animations to teach intracellular signal transduction mechanisms: taking the arrows out of cells. AB - Traditional methods of teaching intracellular biological processes and pathways use figures or flowcharts with the names of molecules linked with arrows. Many veterinary students, presented with such material, simply memorize the names or chemical structures of the molecules and are then likely to forget the material once the examination is completed. To address this problem, the authors designed, created, and field-tested new teaching media that incorporate realistic three dimensional (3D) animations depicting the dynamic changes that occur in intracellular molecules during cellular activation. Testing found that veterinary students taught using traditional teaching media (e.g., lectures, handouts, textbooks) are proficient in memorizing the names and order of intracellular molecules but unable to appreciate the interactions between these elements or their spatial relationships within cells. In contrast, more than 90% of veterinary students taught using 3D animations not only recall the facts about the intracellular elements but also develop accurate mental images of the interactions among these molecules and their spatial relationships. These findings strongly suggest that the comprehension of complex biological processes by veterinary students can be enhanced by the use of dynamic 3D depictions of these processes in the classroom. PMID- 15834825 TI - Integrating a bovine rectal palpation simulator into an undergraduate veterinary curriculum. AB - There are problems associated with teaching bovine rectal palpation to undergraduate veterinary students. The students need opportunities to examine enough cows to develop the required skills, but increasing student numbers and limitations on access to cows have made this more and more difficult to achieve. A virtual reality-based teaching tool, the Bovine Rectal Palpation Simulator, has been developed as a supplement to existing training methods. The student palpates computer generated virtual models of the bovine reproductive tract while interacting with a haptic (touch feedback) device. During training sessions, the instructor follows the student's actions inside the virtual cow on the computer screen and gives instruction. A trial integration of the simulator into the fourth-year bovine reproduction course was undertaken at the University of Glasgow Veterinary School during the 2003/2004 academic year. Students were offered two training sessions, and feedback was gathered using questionnaires. In the first session, all students were taught a range of basic skills using a standardized teaching protocol. The second training session was customized to each student's learning needs and included practice in dealing with a range of on farm scenarios. Student feedback indicated that the training had been useful for learning various aspects of bovine rectal palpation and provided information that helped in the further development of the simulator as a teaching tool. PMID- 15834826 TI - Web-based technology: its effects on small group "problem-based learning" interactions in a professional veterinary medical program. AB - The objective of this investigation was to ascertain whether and how the introduction of a new technology (WebCT) influenced faculty teaching styles while facilitating small group problem-based learning (PBL) sessions in a professional veterinary medical (PVM) program. The following questions guided the study: (1) How does the use of technology affect faculty teaching behaviors? (2) Do the facilitators' interactions with WebCT technology change over the course of one semester? (3) What is the perceived impact of WebCT on facilitators' role in PBL? The study employed a combination of qualitative (case study) and semi quantitative (survey) methods to explore these issues. Nine clinical sciences faculty members, leading a total of six PBL groups, were observed over the course of an academic semester for a total of 20 instructional sessions. The qualitative data gathered by observing faculty as they facilitated PBL sessions yielded three major themes: (1) How do PBL facilitators adapt to the addition of WebCT technology? (2) Does this technology affect teaching? and (3) How do PBL facilitators interact with their students and each other over the course of a semester? No direct evidence was found to suggest that use of WebCT affected teaching behaviors (e.g., student-centered vs. teacher-centered instruction). However, all facilitators showed a moderate increase in comfort with the technology during the semester, and one participant showed remarkable gains in technology skills. The teaching theme provided insight into how facilitators foster learning in a PBL setting as compared to a traditional lecture. A high degree of variability in teaching styles was observed, but individuals' styles tended to remain stable over the course of the semester. Nevertheless, all facilitators interacted similarly with students, in a more caring and approachable manner, when compared to the classroom or clinic atmospheres. PMID- 15834827 TI - Online small animal case simulations, a.k.a. the Virtual Veterinary Clinic. AB - Six years of experience at Washington State University's College of Veterinary Medicine using an online, asynchronous, problem-solving small animal case simulation tool are presented. Student satisfaction with the online problem solving experience was high, and the majority of students felt that the experience was realistic and worthwhile. The effort required to keep the experience realistic required a large investment of time by instructors, which ultimately led to discontinuation of the online course. A summary of features to consider when creating case simulations is presented. PMID- 15834828 TI - ATLes: the strategic application of Web-based technology to address learning objectives and enhance classroom discussion in a veterinary pathology course. AB - A case-based program called ATLes (Adaptive Teaching and Learning Environments) was designed for use in a systemic pathology course and implemented over a four year period. Second-year veterinary students working in small collaborative learning groups used the program prior to their weekly pathology laboratory. The goals of ATLes were to better address specific learning objectives in the course (notably the appreciation of pathophysiology), to solve previously identified problems associated with information overload and information sorting that commonly occur as part of discovery-based processes, and to enhance classroom discussion. The program was also designed to model and allow students to practice the problem-oriented approach to clinical cases, thereby enabling them to study pathology in a relevant clinical context. Features included opportunities for students to obtain additional information on the case by requesting specific laboratory tests and/or diagnostic procedures. However, students were also required to justify their diagnostic plans and to provide mechanistic analyses. The use of ATLes met most of these objectives. Student acceptance was high, and students favorably reviewed the online ''Content Links'' that made useful information more readily accessible and level appropriate. Students came to the lab better prepared to engage in an in-depth and high-quality discussion and were better able to connect clinical problems to underlying changes in tissue (lesions). However, many students indicated that the required time on task prior to lab might have been excessive relative to what they thought they learned. The classroom discussion, although improved, was not elevated to the expected level most likely reflecting other missing elements of the learning environment, including the existing student culture and the students' current discussion skills. This article briefly discusses the lessons learned from ATLes and how similar case-based exercises might be combined with other approaches to enhance and enliven classroom discussions in the veterinary curriculum. PMID- 15834829 TI - Tablet computers in the veterinary curriculum. AB - Tablet computers offer a new method of information management in veterinary medical education. With the tablet computer, students can annotate class notes using electronic ink, search for keywords, and convert handwriting to text as needed. Additional electronic learning resources, such as medical dictionaries and electronic textbooks, can be readily available. Eleven first-year veterinary students purchased tablet computers and participated in an investigation of their working methods and perceptions of the tablet computer as an educational tool. Most students found the technology useful. The small size and portability of the tablet allowed easy transport and use in a variety of environments. Most students adapted to electronic notetaking by the second week of classes; negative experiences with the tablet centered on a failure to become comfortable with taking notes and navigating on the computer as opposed to writing and searching on paper. A few performance-related problems, including short battery life, were reported. Tablet software allowed conversion of faculty course notes from a variety of original formats, meaning that instructors could maintain their original methods of note preparation. Adopting a consistent naming convention for files helped students to locate the files on their computers, and smaller file sizes helped with computer performance. Collaboration between students was fostered by tablet use, which offers possibilities for future development of collaborative learning environments. PMID- 15834830 TI - Implementing a personal digital assistant (PDA) program in a veterinary college curriculum. PMID- 15834831 TI - Handheld computers in veterinary medical education: a view from human medical education. AB - Handheld computers are widely used in clinical practice, and their use in both human medical education and veterinary medical education is increasing, especially, for the former, in activities involving point-of-care access. This article references the insights that can be obtained from the usage and activities that are gaining a strong foothold in human medical education. Handheld computer technology gives students access to a large and changing knowledge base for clinical practice, especially when they are geographically dispersed. Differences in use between education and practice largely relate to the importance clinicians place on patient information. Student use focuses on progress mapping and ready access to clinical reference material. Suggestions are made for future use in medical education. PMID- 15834832 TI - Using a wireless response system to enhance student learning. PMID- 15834833 TI - Exploring student attitudes to directed self-learning online through evaluation of an Internet-based biomolecular sciences resource. AB - RATIONALE FOR THE STUDY: In 2000, funding was awarded by the University of Glasgow's Learning and Teaching Development Fund (L&TDF) for the authors to develop an interactive, online learning resource for veterinary biomolecular sciences teaching. This course is a core component of the veterinary undergraduate curriculum at the university. Evaluations were carried out to gauge students' experiences of using the resource as a basis for exploring students' attitudes toward online, independent learning. METHODOLOGY: Peers were asked to review the design and content of four modules, also evaluated by students using questionnaires and focus group discussions. Additionally, students were observed using the modules. Both first-year students and second-year direct-entry students (i.e., students entering the veterinary program with advanced training) participated in the evaluation, which allowed for some comparison between the groups. One cohort used the modules independently, and their responses were compared with the cohorts that used the modules in scheduled classes. RESULTS AND CONCLUSIONS: The evaluations indicate that this is a useful resource that could act as a template for other courses within the veterinary undergraduate curriculum, particularly for learning of basic sciences. On average, first-year and timetabled students rated the program more highly overall, rated the program more highly in relation to previous instruction, and rated tutor presence as more important than second-year direct-entry and independent students did. The lower rating given to tutor presence by second-year direct-entry and independent students indicates that they are more confident using the modules without tutor supervision. PMID- 15834834 TI - Student assessment of the educational benefits of using a CD-ROM for instruction of basic surgical skills. AB - RATIONALE FOR STUDY: At Texas A&M University, introductory-level surgical lecture and laboratory notes were converted to a CD-ROM format that included illustrative photographs as well as instructional videos demonstrating the basic surgical skills that all students were required to master. The CD-ROM was distributed to all students in place of traditional paper notes in the second-year surgical class in the professional veterinary curriculum. The study reported here was designed to evaluate the educational benefits of the use of the CD-ROM in place of traditional paper notes by examining the attitudes and practices of students before and after exposure to the CD-ROM format. METHODOLOGY: An anonymous survey was distributed to students in the second-year introductory surgery course on the first day of class and again on the last day of class. Responses to questions were tabulated, response frequencies determined, and Chi-square analysis performed to determine differences between initial and final responses. RESULTS: On the final survey, 89 per cent of students responded that the instructional videos definitely helped them prepare for the laboratory, and 77 per cent responded that they were more likely to practice techniques learned from the CD ROM videos than those learned from traditional study materials. The majority of students believed that the CD-ROM improved both the course (60 per cent) and their learning experience (62 per cent) as compared to traditional paper notes. CONCLUSIONS: Including instructional videos on the CD-ROM enhanced the educational experience of the students by promoting preparedness for laboratories and promoting practice of techniques learned from the videos outside of the laboratory. PMID- 15834835 TI - A quiz becomes a multidirectional dialogue with Web-based instructional tools for an anatomical pathology rotation. AB - Senior veterinary students in the Iowa State University College of Veterinary Medicine (ISU CVM) participate in clinical rotations, among them a two-week necropsy rotation. The students have access to the rotation syllabus on the ISU CVM intranet site. To promote rapid comprehension of necropsy protocol, students completed a pre-exam on the syllabus. This exercise evolved from a paper quiz to an online pre-exam, using course management software to improve use of class time, increase feedback, and shift the focus to acquisition of knowledge. The students were encouraged to work collaboratively on the pre-exam and could make repeated attempts. We predicted that professional students would make multiple attempts at the pre-exam until the desired score was attained. This exercise achieves multiple goals. First, the exam encourages early review of necropsy protocol. Second, use of WebCT allows for instant, automatic, and consistent feedback from the instructor, reducing redundancy while improving the quality of communication between student and instructor and thus using faculty time more efficiently. The instructor can quickly identify and rectify common misunderstandings through this interface. Third, by allowing discussion and repeated attempts, we can ensure that there is less pressure associated with the exam. Statistical analysis of the students' performance supports the prediction that students would repeat the exam until the desired score was achieved. Subjectively, as a result of implementation of an online pre-exam, the instructor has observed students to be more engaged with the material at an earlier point in the rotation. PMID- 15834837 TI - [Somatization -- conversion -- dissociation: strategies for behavior therapy]. AB - Modern cognitive behavioral approaches for the treatment of patients with medically unexplained somatic symptoms have been developed on the basis of the classification systems DSM-IV and ICD-10. These systems define somatoform disorders as a homogeneous clinical group. Behavior therapy has additionally developed vicious circle models specifying etiological, triggering and maintaining factors. Treatment goals and strategies can be derived directly from these models. The main components are: (1) motivation of patients to accept the psychotherapeutic approach; (2) introduction of alternative explanations of the symptoms on the basis of both biomedical as well as psychosocial mechanisms; (3) evaluation of the new explanations by patient and therapist; (4) reduction of avoidance and inadequate illness behaviour. Health economical aspects are particularly important because patients with somatoform disorders tend to overuse medical services and are thus considered an expensive problem group for the health system. PMID- 15834838 TI - [Results of inpatient psychiatric and psychotherapeutic treatment of patients with schizophrenia, schizoaffective and other psychotic disorders]. AB - OBJECTIVES: At the Tiefenbrunn hospital, patients suffering from severe psychiatric and psychosomatic disorders are treated. The treatment concept is psychodynamically oriented. Psychopharmacological treatments are included if necessary. This article reports the results of the treatment of a sample of patients with schizophrenic and other psychotic disorders (ICD-10: F20-F25). METHODS: An unselected sample of N = 33 patients was studied upon admission and before discharge with a set of routinely administered standardized instruments. RESULTS: According to the data, all of these patients were severely disturbed: Upon admission to the clinic, they showed high pathological values in the Beeintrachtigungs- Schwere-Score (degree of impairment score; Schepank 1995), the Symptom Checklist SCL-90-R (Franke 1995), the Inventory of Interpersonal Problems (Horowitz et al. 1994), as well as a high degree of comorbid disorders. A comparison of the data at admission and at discharge shows that these patients improved significantly and substantially concerning symptoms, interpersonal problems, contentedness with life and individually formulated target problems. DISCUSSION: The results of the present study are discussed with regard to both the treatment of psychotic patients and the results of other studies. PMID- 15834839 TI - [Utilization of the health care system due to somatoform complaints -- results of a representative survey]. AB - OBJECTIVES: Utilization of the health care system is not determined by the presence of an organic illness, but by a multitude of interacting factors. This applies particularly to somatoform complaints and disorders, in the context of which the health care system is used very intensively and inadequately in relation to the disorder. The present paper summarizes the frequency of somatoform complaints in the general population and health care utilization due to these complaints. METHODS: On the basis of a survey carried out in 2002 for a representative population of 2089 persons (aged 14 to 92 years), the prevalence of somatoform complaints during a period of 2 years was determined. In this survey, physical complaints were defined as being somatoform when they seriously affected the person's well-being but exhibited a disparity between objective diagnostic findings and subjective experience. The psychosocial aspects were also determined. When somatoform complaints were identified, the specific utilization of the healthcare system for them was investigated. RESULTS: Somatoform complaints are very frequent in Germany: 8.3% of the population claim to suffer from clinically relevant somatoform disorders that fully meet the criteria mentioned above. The health care system is used heavily for these complaints. Within the last two years, each test person suffering from somatoform disorders consulted a physician a mean 18 times and was incapable of working for a mean of 20 days. 16% of these persons were hospitalized, 9 % received treatment at a health resort, 9 % retired, etc. DISCUSSION: The considerable utilization of health care services is an essential element of somatoform pathology and has immense medico-political relevance. The most important prerequisite for handling the 'commitment to health care system' symptom is thorough training of all physicians in the basics of somato-psycho-social medicine. This will be an important measure for cost reduction in the health care system. PMID- 15834840 TI - [Oxytocin, a mediator of anti-stress, well-being, social interaction, growth and healing]. AB - The neuroendocrine and physiological systems related to pain and stress have long been subjected to study. More recently, the corresponding systems promoting anti stress and restoration have also come into focus. It is not only important to investigate the mechanisms underlying disease but also to examine the physiological and psychological mechanisms which protect and heal the body and soul. The nonapeptide oxytocin, originally known to stimulate labour and milk ejection, appears to play an important role in this regard. Oxytocin can induce anti-stress-like effects such as reduction of blood pressure and cortisol levels. It increases pain thresholds, exerts an anxiolytic-like effect and stimulates various types of positive social interaction. In addition, it promotes growth and healing. Repeated exposure to oxytocin causes long-lasting effects by influencing the activity of other transmitter systems, a pattern which makes oxytocin potentially clinically relevant. Oxytocin can be released by various types of non noxious sensory stimulation, for example by touch and warmth. Ingestion of food triggers oxytocin release by activation of vagal afferents. Most likely, oxytocin can also be released by stimulation of other senses such as olfaction, as well as by certain types of sound and light. In addition, purely psychological mechanisms may trigger the release of oxytocin. This means that positive interaction involving touch and psychological support may be health-promoting. The social interaction of daily life, as well as a positive environment, continuously activate this system. In addition, various types of psychotherapy involving transfer of support, warmth and empathy are likely to induce similar effects, which thus contribute to the positive effects of these kinds of therapies. PMID- 15834841 TI - Quantification of imidacloprid toxicity to avocado thrips, Scirtothrips perseae Nakahara (Thysanoptera: Thripidae), using a combined bioassay and ELISA approach. AB - A competitive ELISA (enzyme-linked immunosorbent assay) technique was evaluated for quantifying titres of imidacloprid in homogenates of leaf discs sampled from avocado plants treated with systemic applications of imidacloprid 240 g litre(-1) SC (Admire). Matrix effects were evident with undiluted leaf tissue homogenates, but these were effectively eliminated by dilution of homogenates in water. In a field trial conducted in a commercial nursery, there was an excellent correlation between imidacloprid residues within leaves and avocado thrips (Scirtothrips perseae Nakahara) mortality. However, with subsequent sampling over a 2-month period, the relationship between mortality and apparent imidacloprid concentration became less robust, suggesting that the material was being degraded within the plant to non-toxic metabolites. Nevertheless, assessments of thrips mortality on leaves that had been recently treated with imidacloprid established a lower threshold of activity for imidacloprid residues of 6 ng cm(-2) leaf. Limitations on the use of ELISA to quantify the impact of systemic insecticides on pest populations are discussed. PMID- 15834842 TI - Correlation of a resistance-associated Rdl mutation in the German cockroach, Blattella germanica (L), with persistent dieldrin resistance in two Danish field populations. AB - Danish Blattella germanica (L) populations carry the resistance-associated mutation A302S within the Resistance to dieldrin (Rdl) gene. The mutation has remained in field populations long after the discontinuation of dieldrin for cockroach control. The mutation has also persisted in our laboratory strains with high and intermediate frequencies for more than 8 years without selection. The toxicity of dieldrin was tested by topical application to male cockroaches in the susceptible strain DPIL-SUS and two field strains, Zo960302 and Su960304, which were 1270- and 15-fold resistant to dieldrin at LD50. We report the sequencing of exon 7 of the B. germanica Rdl gene and the finding of the putative resistance associated A302S mutation. We have developed and implemented a PCR-based diagnostic method with the detection of a restriction endonuclease polymorphism, which allows for easy discrimination of susceptible, resistant and heterozygote genotypes. The frequency of the resistance-associate allele A302S was 0.97 and 0.38 in the Zo960302 and Su960304 populations, respectively. The cockroach Rdl A302S allele confers high dieldrin resistance in homozygotes and intermediate resistance in heterozygotes, and its presence is responsible for the persisting dieldrin resistance in Danish populations of B. germanica. PMID- 15834843 TI - The effect of nabilone on neuropsychological functions related to driving ability: an extended case series. AB - The primary goal of this prospective extended case series was to obtain the first data about the potential influence of nabilone intake on driving ability related neuropsychological functions. Six patients were investigated within a placebo controlled, double-blind crossover study of this synthetic cannabinoid (2 mg/day) in patients with multiple sclerosis and spasticity associated pain. Five neuropsychological functions (reaction time, working memory, divided attention, psychomotor speed and mental flexibility) were assessed. No indication was found of a deterioration of any of the five investigated neuropsychological functions during the 4-week treatment period with nabilone. PMID- 15834844 TI - Dysregulation of hedgehog signalling predisposes to synovial chondromatosis. AB - Synovial chondromatosis is a condition affecting joints in which metaplastic cartilage nodules arise from the synovium, causing pain, joint dysfunction, and ultimately joint destruction. Because dysregulation of hedgehog signalling is a feature of several benign cartilaginous tumours, expression of the hedgehog target genes PTC1 and GLI1 was examined in this study in samples from human synovial chondromatosis. Significantly higher expression levels were found in synovial chondromatosis than in the synovium, from which it arises. To determine if hedgehog-mediated transcription predisposes to synovial chondromatosis, the extra-toes mutant mouse, which harbours a heterozygous mutation in the hedgehog transcriptional repressor, Gli3, resulting in decreased expression of Gli3 protein, was studied. The extra-toes mutant mouse has a phenotype consistent with overactive hedgehog signalling, suggesting that Gli3 acts as a transcriptional repressor of limb development. Eighty-five per cent of Gli3 mutant mice developed synovial chondromatosis at 18 months of age, compared with 30% of wild-type littermates (p < 0.05). Three of the ten Gli3 mutant mice treated with triparanol, which blocks hedgehog signalling upstream of the Gli transcription factors, developed synovial chondromatosis, compared with eight of ten control mice. These data demonstrate that hedgehog signalling plays an important role in the development of synovial chondromatosis and suggest that blockade of hedgehog signalling may be a potential treatment for this disorder. PMID- 15834845 TI - Denaturation of metalloproteins with EDTA to facilitate enzymatic digestion and mass fingerprinting. AB - Metal ions bound to a protein often stabilize tertiary and/or quaternary structure. Consequently, the digestion of metalloproteins that precedes analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is frequently incomplete. It is demonstrated that ethylenediaminetetraacetic acid (EDTA) successfully destabilizes metalloprotein structure and thereby facilitates tryptic digestion and protein identification. PMID- 15834846 TI - Quantification of sifuvirtide in monkey plasma by an on-line solid-phase extraction procedure combined with liquid chromatography/electrospray ionization tandem mass spectrometry. AB - A simple, automated and rapid method has been developed for the determination of a novel antiviral peptide sifuvirtide in monkey plasma. Raw plasma samples were directly loaded onto an on-line solid-phase extraction (SPE) column, which removes the time-consuming and laborious sample pretreatment. Following a timed valve-switching event, the analyte was eluted on-line to a reversed-phase high performance liquid chromatography (RP-HPLC) column and subsequently introduced into a linear ion trap mass spectrometer, LTQ-MS, via an electrospray ionization (ESI) interface. The multiply charged peptides were specified and quantitatively analyzed using selective reaction monitoring (SRM). A highly pure four iodine sifuvirtide was synthesized using an optimized iodogen method and proved to be a suitable internal standard (IS). A single analysis run takes about 18 min. Validation of the method demonstrated that the linear calibration curves covered the range of 4.88-5000 ng/mL, and the correlation coefficients were above 0.9923. The limit of detection (LOD) with the signal-to-noise (S/N) ratio higher than 12 was calculated as 1.22 ng/mL. The intra- and inter-batch precisions were less than 12.7% and 9.1%, and the mean accuracy ranged from -5.2% to 3.6%, respectively. Any carry-over effect from the system was negligible. In a pharmacokinetic (PK) study of sifuvirtide after a single intravenous or subcutaneous dose in monkeys, the on-line SPE-LC/MS/MS system was successfully utilized to determine hundreds of samples with only one extraction column, which indicated the feasibility and the reliability of this method for application in preclinical and clinical PK studies of peptide drugs. PMID- 15834847 TI - A highly diastereomerically enriched, silyl-substituted alkyl lithium, configurationally stable at room temperature. PMID- 15834848 TI - Stretched gelatin gels as chiral alignment media for the discrimination of enantiomers by NMR spectroscopy. PMID- 15834849 TI - Coordination chemistry of the hexavacant tungstophosphate [H2P2W12O48]12- with Fe(III) ions: towards original structures of increasing size and complexity. PMID- 15834850 TI - Total chemical synthesis and X-ray crystal structure of a protein diastereomer: [D-Gln 35]ubiquitin. PMID- 15834851 TI - 2D NMR characterization of the La@C82 anion. PMID- 15834852 TI - Assembly of the complete eight-base artificial genetic helix, xDNA, and its interaction with the natural genetic system. PMID- 15834853 TI - Evidence for carbon-carbon Meisenheimer-Wheland complexes between superelectrophilic and supernucleophilic carbon reagents. PMID- 15834854 TI - Identifying and distinguishing cases of parkinsonism and Parkinson's disease using ICD-9 CM codes and pharmacy data. AB - Administrative databases have the potential to assess quality and cost of care for parkinsonism and Parkinson's disease. However, the validity of findings is limited by our understanding of how cases are identified. Patient records listing International Classification of Diseases, Version 9, Clinical Modification (ICD-9 CM) codes for parkinsonism (n = 2,076) and dopaminergic medications (n = 2,798) were pulled from fiscal years 1999 to 2001 for patients in the Pacific Northwest Veterans Administration. Samples of these records (n = 397) and records without these ICD-9 CM codes (n = 500) were reviewed, and clinical data were extracted. The accuracy of administrative data to identify and distinguish between Parkinson's disease and parkinsonism was calculated. A total of 37.9% of parkinsonism cases were detected using pharmacy data and ICD-9 CM codes compared to 18.7% by using ICD-9 CM codes alone. The ICD-9 CM code for paralysis agitans (332.0) did not distinguish between probable Parkinson's disease and other causes of parkinsonism, whereas the ICD-9 CM code for degenerative basal ganglia disorder (333.0) predicted having secondary parkinsonism (odds ratio [OR] = 5.0) as well as dopa-responsiveness in patients without secondary parkinsonism (OR = 4.5). Administrative data are limited in the ability to identify parkinsonism. The ICD-9 CM code, 332.0, which is generally considered the code to identify Parkinson's disease, did not distinguish between parkinsonism and Parkinson's disease. PMID- 15834855 TI - Association between male gender and pediatric essential tremor. AB - Approximately 5% of new essential tremor (ET) cases arise during childhood. The goal of the current report was to examine the possible association between male gender and pediatric ET, using data from 95 pediatric ET cases seen at three medical centers (two in the United States and one in Spain). The odds of ET in our sample of cases were threefold higher in boys compared to girls. Whether this association between male gender and pediatric ET represents a selection bias or a true gender-mediated biological difference in disease expression is not known, although some data support the latter possibility. PMID- 15834856 TI - Anticancer multidrug resistance mediated by MRP1: recent advances in the discovery of reversal agents. AB - Multidrug resistance protein 1 (MRP1) belongs to the ATP-binding cassette (ABC) transporter family. It is able to transport a broad range of anticancer drugs through cellular membranes, thus limiting their antiproliferative action. Since its discovery in 1992, MRP1 has been the most studied among MRP proteins, which now count nine members. Besides the biological work, which targets structure elucidation, binding sites location, and mode of action, most efforts have been focused on finding molecules which act as MRP1 inhibitors. In this review, we attempt to summarize and highlight studies dealing with modulators of MRP1 mediated multidrug resistance (MDR), which have been accomplished in the last 5 years. The reported MRP1 inhibitors are discussed according to their chemical class. Finally, we try to bring information on structure-activity relationship (SAR) aspects and how modulators might interact with MRP1. This study may facilitate the rational design of future modulators of MDR. PMID- 15834857 TI - Increased tau burden in the cortices of progressive supranuclear palsy presenting with corticobasal syndrome. AB - The objective of this study is to better define the pathological characteristics of pathologically proven progressive supranuclear palsy (PSP) presenting with the corticobasal syndrome (CBS). PSP is characterized by early falls, vertical supranuclear ophthalmoplegia, and axial rigidity, whereas asymmetric limb features, including rigidity, bradykinesia, apraxia, alien limb phenomena, and cortical sensory loss are characteristic of CBS. We investigated clinicopathological characteristics of 5 cases of PSP that presented with CBS (CBS-PSP). Comprehensive pathological analysis was undertaken to determine the presence of concomitant pathological processes as well as quantitative tau burden in cortical regions of CBS-PSP, compared with 8 typical PSP cases (Typ-PSP). The clinical features in the CBS-PSP cases included asymmetrical features, apraxia, alien limb phenomena, and progressive aphasia. All cases had Parkinsonism, and vertical supranuclear ophthalmoplegia was noted in all but 1 case of CBS-PSP. Secondary neuropathological diagnoses included argyrophilic grain disease (AGD) in 1 of the 8 cases of Typ-PSP, whereas Alzheimer's disease (AD), Lewy body disease, AGD, and vascular disease was found in 3 cases of CBS-PSP. Image analysis of cortical tau burden performed in 8 Typ-PSP and 3 CBS-PSP cases revealed a significant increased tau burden in mid-frontal and inferior-parietal cortices in the CBS-PSP cases. This study demonstrates that when PSP presents as CBS, it is most likely due to either a concurrent cortical pathology from a secondary process such as AD or from the primary pathology of PSP extending into cortical areas that are primarily and commonly affected in CBD. PMID- 15834858 TI - Atypical Hallervorden-Spatz disease with preserved cognition and obtrusive obsessions and compulsions. AB - We describe the case of an adult female with Hallervorden-Spatz disease (HSD), "eye-of-the-tiger" sign on cranial magnetic resonance imaging scan, and two mutations in the pantothenate kinase 2 (PANK2) gene. Symptomatic presentation included stuttering dysarthria, dystonic posturing, increased limb and axial muscle tone, choreoathetosis, stereotyped motor behaviors, and obsessive compulsive symptomatology since adolescence. Extensive neuropsychological testing at 40 and 44 years of age revealed a relatively normal IQ and stable cognitive pattern overall. This case demonstrates that HSD patients who survive into middle age should not be assumed to have a progressive dementia. In such cases, atypical behavioral problems such as persistent obsessions and compulsions may be present instead. PMID- 15834859 TI - Glutathione-S-transferase-1 and interleukin-1beta gene polymorphisms in Japanese patients with Parkinson's disease. AB - We studied genetic polymorphisms in the glutathione-S-transferase-1 (GST-1) gene region and the interleukin-1beta (IL-1beta) promoter region in patients with Parkinson's disease (PD, n = 361), as well as controls (n = 257). Although we have confirmed the previous results, in a larger sample, that the IL-1beta genotype has affected the age at onset of PD patients, no contribution of the GST 1 gene polymorphism was observed in the allele frequency or the onset age of the disease in Japanese persons. PMID- 15834861 TI - Anterior cingulate cortex: an fMRI analysis of conflict specificity and functional differentiation. AB - In this event-related functional magnetic resonance imaging (fMRI) study, we provide evidence that the role of the anterior cingulate cortex (ACC) in cognitive control may not be unitary, as the responses of different ACC subregions vary depending upon the nature of task-irrelevant information. More specifically, using the color-word Stroop task (congruent, incongruent, and neutral trial types), we examined the degree to which increases in neural activity within ACC are specific to conditions of conflict, as posited by the conflict monitoring theory (Botvinick et al. [1999]: Rev Neurosci 10:49-57; Carter et al. [1998]: Science 280:747-749). Although incongruent and congruent trials both involve two competing sources of color information (color word and ink color), only incongruent trials involve a direct conflict between task relevant and task-irrelevant information. Although the anterior division of the ACC rostral zone exhibited conflict specific increases in neural activity (i.e., incongruent > congruent = neutral), the posterior division exhibited a more generalized pattern, increasing whenever the task-irrelevant information was color related, regardless of whether it was conflicting (i.e., incongruent and congruent > neutral). Our data thus suggest a possible functional differentiation within the ACC. As such, it is unlikely that the role of the ACC in cognitive control will be able to be accommodated by a single unifying theory. PMID- 15834860 TI - Using automated morphometry to detect associations between ERP latency and structural brain MRI in normal adults. AB - Despite the clinical significance of event-related potential (ERP) latency abnormalities, little attention has focused on the anatomic substrate of latency variability. Volume conduction models do not identify the anatomy responsible for delayed neural transmission between neural sources. To explore the anatomic substrate of ERP latency variability in normal adults using automated measures derived from magnetic resonance imaging (MRI), ERPs were recorded in the visual three-stimulus oddball task in 59 healthy participants. Latencies of the P3a and P3b components were measured at the vertex. Measures of local anatomic size in the brain were estimated from structural MRI, using tissue segmentation and deformation morphometry. A general linear model was fitted relating latency to measures of local anatomic size, covarying for intracranial vault volume. Longer P3b latencies were related to contractions in thalamus extending superiorly into the corpus callosum, white matter (WM) anterior to the central sulcus on the left and right, left temporal WM, the right anterior limb of the internal capsule extending into the lenticular nucleus, and larger cerebrospinal fluid volumes. There was no evidence for a relationship between gray matter (GM) volumes and P3b latency. Longer P3a latencies were related to contractions in left temporal WM, and left parietal GM and WM near the interhemispheric fissure. P3b latency variability is related chiefly to WM, thalamus, and lenticular nucleus, whereas P3a latency variability is not related as strongly to anatomy. These results imply that the WM connectivity between generators influences P3b latency more than the generators themselves do. PMID- 15834862 TI - Association testing with Mendel. AB - This report presents an overview of association testing strategies from a user's perspective, with particular attention to the capabilities of the computer program Mendel. Association testing is driven by the nature of the study sample, the nature of the disease trait, and the kind of markers employed. The practicing statistician must also choose whether to conduct parametric or nonparametric tests. Because of the complexities involved, Mendel offers users several analysis options. The different options are tied together by shared input and output conventions and a shared language for defining models. Mendel also features new statistics and theory found in no other genetics software. The most important innovations include: association testing by penetrance estimation, expansion of matched-pair designs to permutation unit designs, and a rigorous implementation of the measured genotype approach for quantitative trait loci. This report explains how Mendel imputes allele counts and conducts both asymptotic and permutation tests in the measured genotype framework. PMID- 15834863 TI - Optic radiation changes after optic neuritis detected by tractography-based group mapping. AB - Postmortem data suggest that trans-synaptic degeneration occurs in the lateral geniculate nucleus after optic nerve injury. This study investigated in vivo the optic radiations in patients affected by optic neuritis using fast marching tractography (FMT), a diffusion magnetic resonance imaging (MRI) fiber tracking method, and group mapping techniques, which allow statistical comparisons between subjects. Seven patients, 1 year after isolated unilateral optic neuritis, and ten age and gender-matched controls underwent whole-brain diffusion tensor MR imaging. The FMT algorithm was used to generate voxel-scale connectivity (VSC) maps in the optic radiations in each subject in native space. Group maps of the left and right optic radiations were created in the patient and control group in a standardized reference frame using statistical parametric mapping (SPM99). The reconstructed optic radiations in the patient group were localized more laterally in the posterior part of the tracts and more inferiorly than in the control group. Patients showed reduced VSC values in both tracts compared with controls. These findings suggest that the group mapping techniques might be used to assess changes in the optic radiations in patients after an episode of optic neuritis. The changes we have observed may be secondary to the optic nerve damage. PMID- 15834864 TI - Epidemiological and phylogenetic evidence for patient-to-patient hepatitis C virus transmission during sclerotherapy of varicose veins. AB - The aim of this study was to provide evidence for patient-to-patient nosocomial hepatitis C virus (HCV) transmission during sclerotherapy of varicose veins. Forty-three patients who had evidence of current infection by genotype 2 HCV have had sclerotherapy by the same physician. Based on this observation, a detailed epidemiological questionnaire on risk factors for HCV in genotype 2 infected patients was conducted. Seventeen sequences in the hypervariable region 1 (HVR1) of the HCV genome obtained from 17 HCV RNA positive patients with a past history of sclerotherapy, were compared with 17 sequences derived from genotype 2 patients with no past history of sclerotherapy, and with 25 sequences sampled from GenBank. Two hundred seven genotype 2 HCV infected patients were included. The main risk factors for HCV infection were transfusion (n = 76), drug use (n = 6), and sclerotherapy of varicose veins (n = 62 including 43 (20.8%) by the same physician), other or unknown (n = 76). These sclerotherapy sessions were carried out in the 1980s for many years. Five of these 43 patients had jaundice within a few weeks after a sclerotherapy session. Sequence analysis of HVR1 from 17 patients who had sclerotherapy by the same physician revealed that they were all infected with HCV genotype 2c. The phylogenetic tree indicated clustering of the patients with a past history of sclerotherapy. The method by which infection was likely to have been transmitted was probably the use of a single vial for multiple patients. This study provides strong evidence that sclerotherapy of varicose veins is a risk factor for HCV infection. PMID- 15834865 TI - Clinical outcome after 4 years follow-up of HIV-seropositive subjects with incomplete virologic or immunologic response to HAART. AB - The duration of the clinical, virologic, and immunologic response to HAART, is not well defined. In this observational multi-center study 2,143 patients were enrolled classified according to virologic suppression (<500 cp/ml) and immune recovery (>100 CD4+ cells/mul from baseline) at month 12 of HAART as complete responders, virologic only responders, immunologic only responders and non responders. Kaplan Meyer curves, multivariate and politomous regression analysis were used. Complete responders patients were 781 (36.4%), immunologic only responders 441 (20.6%), virologic only responders 336 (15.7%), and non-responders 585 (27.3%). Using multivariate analysis, being antiretroviral-naive increased the probability of having both a virologic only or a complete response and reduced the probability of an immunologic only response (P < 0.001 for all tests). Older age was associated directly with a virologic only response and inversely associated with an immunologic only response (P = 0.027 and P = 0.035, respectively). Using politomous analysis, patients baseline HIV-RNA level more than 5 log cp/ml had a 1.9-fold higher probability of an immunologic response than of a complete response (P = 0.001). After 4 years, the clinical progression rate was six times greater in non-responders, 1.9 times greater in virologic only responders, and 2.3 times greater in immunologic only responders than for responders. However, patients with virologic only response or with immunologic only response had a significantly reduced risk for clinical progression than non responders (P < 0.001). After 4 years of HAART, the risk of clinical progression in patients with immunologic only or virologic only response is low but still higher than in complete responder patients. PMID- 15834866 TI - Detection of human herpesvirus 7 DNA in peripheral blood reflects mainly CD4+ cell count in patients infected with HIV. AB - The opportunistic behavior and the potential interactions of human herpesvirus 7 (HHV-7) with human immunodeficiency virus (HIV)-1 in HIV-1-infected patients were investigated in comparison with HHV-6, another human roseolovirus. Roseolovirus DNAs were detected and quantified in peripheral blood mononuclear cells (PBMCs) from 198 HIV-seronegative healthy blood donors, 38 HIV-1-infected patients classified as long-term non-progressors, and 99 HIV-1-infected patients classified as progressors. The rate of HHV-7 DNA detection was higher in healthy donors (78%) than in long-term non-progressors (47%; P = 0.0003) or in progressors (52%; P < 0.0001). HHV-7 cell load was higher in healthy donors (median: 212 EqCop/10(6) PBMCs) and in long-term non-progressors (median: 105 EqCop/10(6) PBMCs) than in progressors (median: 48 EqCop/10(6) PBMCs; P < 0.0001 and P = 0.015, respectively). Among progressors, HHV-7 detection was correlated positively with the CD4(+) T-lymphocyte count (P = 0.028). Neither HHV-7 detection rate nor cell load was correlated with the HIV-1 plasma load. As a whole, HHV-6 detection rate and cell load were lower than the HHV-7 counterparts, albeit exhibiting similar differences between healthy donors, long-term non progressors, and progressors. In conclusion, HHV-7 infection does not appear to be stimulated by HIV-1 infection, nor interact with it. Rather, HHV-7 detection rate and cell load reflect CD4(+) T-lymphocyte count, with higher values in healthy donors and long-term non-progressors than in progressors. PMID- 15834867 TI - Comparison of affinity chromatography and adsorption to vaccinia virus recombinant infected cells for depletion of antibodies directed against respiratory syncytial virus glycoproteins present in a human immunoglobulin preparation. AB - Antibodies directed against human respiratory syncytial virus (HRSV) glycoproteins were depleted from a commercial immunoglobulin preparation (RespiGam) by two different methods. The first method consisted of repeated adsorption of RespiGam to Sepharose beads with covalently bound soluble forms of the two major viral glycoproteins (F or G). The second method consisted of adsorption of immunoglobulins to live cells expressing F or G glycoproteins on their surfaces after infection with vaccinia virus recombinants. While the first method removed efficiently antibodies that reacted with F and/or G glycoproteins by ELISA, it was inefficient in the elimination of anti-HRSV neutralizing antibodies. In contrast, the second method removed efficiently anti-HRSV antibodies that both reacted by ELISA and neutralized virus infectivity. These results confirm that human neutralizing antibodies are directed exclusively against HRSV F and G glycoproteins, and, they raise the possibility that F and G glycoproteins inserted into cell membranes differ antigenically from their soluble forms linked covalently to Sepharose beads. PMID- 15834868 TI - Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS. AB - A new Coronavirus (SARS-CoV) is the aetiological agent of the severe acute respiratory syndrome (SARS). Because of the critical role played by serological assays for SARS diagnosis, an in-house ELISA based on SARS-CoV recombinant antigens was developed. The SARS-CoV nucleocapsid protein (N), three N fragments (N1, N2, and N3) and the intraviral domain of the membrane protein (M2) were cloned and expressed in Escherichia coli as histidine-tagged proteins. Six reference sera from SARS patients were used to detect virus-specific IgG in an ELISA using each recombinant protein as coating antigen. High-titre positive reactions were detected in all SARS positive sera. The specificity of the assay appears to be high as no positive reaction was detected in the sera of 20 healthy subjects and 73 patients with non-SARS, low-tract respiratory infections. Specific hyper-immune sera to SARS-CoV and the recombinant proteins, N, N1, N2, N3, and M2 were also generated in mice and rabbits. The specificity of these sera was confirmed by an immunocytochemical assay on biochips of SARS-CoV infected and uninfected cells. PMID- 15834870 TI - Infection with human T lymphotropic virus type I in organ transplant donors and recipients in Spain. AB - Human T-cell lymphotropic virus (HTLV) antibody screening is not recommended uniformly before transplantation in Western countries. In the year 2001, the first cases of HTLV-I infection acquired through organ transplantation from one asymptomatic carrier were reported in Europe. All three organ recipients developed a subacute myelopathy shortly after transplantation. This report rose the question about whether to implement universal anti-HTLV screening of all organ donors or selective screening of donors from endemic areas for HTLV-I infection should be carried out. A national survey was conducted thereafter in which anti-HTLV antibodies were tested in 1,298 organ transplant donors and 493 potential recipients. None was seropositive for HTLV-I and only one recipient, a former intravenous (i.v.) drug user, was found to be infected with HTLV-II. In a different survey, HTLV screening was conducted in 1,079 immigrants and 5 (0.5%) were found to be asymptomatic HTLV-I carriers. All came from endemic areas for HTLV-I infection. No cases of HTLV-II infection were found among immigrants. These results support the current policy of mandatory testing of anti-HTLV antibodies in Spain only among organ transplant donors coming from HTLV-I endemic areas or with a highly suspicion of HTLV-I infection. PMID- 15834869 TI - Molecular epidemiology of hepatitis B virus in Spain: identification of viral genotypes and prediction of antigenic subtypes by limited sequencing. AB - The hepatitis B virus (HBV) genotypes were studied by a line probe assay (LiPA) and by direct sequencing of a 339 nucleotide fragment from the S region of the viral genome in samples from 269 carriers living in Spain, either native to Spain (231) or immigrants from Africa, Asia, and Eastern Europe (38). The sequences were also used to predict the HBV surface antigen (HBsAg) subtype on the basis of the amino acids specified at selected positions of the HBsAg molecule. Agreement between the two genotyping methods was found in most cases (98.1%) and a HBV genotype could be assigned to all samples. The viral groups D/ayw2 (30.1%), D/ayw3 (28.6%), and A/adw2 (21.2%) were prevalent, with an additional participation of the groups D/ayw4 (4.8%), F/adw4q- (1.9%), A/ayw1 (1.9%), and D/adw3 (0.7%), all of them present among the autochthonous carriers. Strains from genotypes B and C were found exclusively among Chinese immigrants. Genotype E strains were found in immigrants from Central Africa and in one patient native of Spain. Point mutations leading to amino acid changes of residues involved in the expression of the HBsAg subtype determinants were found in 12 samples (4.5%). Some mutations would predict the putative novel genotype-subtype associations A/adw4q+, A/ayr, D/ayr, and E/ayw1, while others would suggest the loss of subtype-specific determinants. The finding of HBV strains characteristic for Africa among the autochthonous carriers confirms the emergence of African HBV strains in Spain. PMID- 15834871 TI - Serological study of the lymphochoriomeningitis virus (LCMV) in an inner city of Argentina. AB - Lymphocytic choriomeningitis virus (LCMV) is the prototype of the family Arenaviridae and is associated with the natural reservoir, Mus domesticus (Md). It causes meningitis and a flu-like illness characterized by malaise, myalgia, retrorbital headache, and photophobia. This study presents the data obtained in a rodent and human serological study during 6 years (1998-2003) in the city of Rio Cuarto, Argentina. Antibodies anti-LCMV were sought by ELISA in rodents and humans. LCMV was found only in Md species in 9.4% of animals. The results also show some seasonal, no significant variations in the prevalence of the infection. Distribution of positive mice was not modified significantly by trapping sites, sex, or age of the animals. The prevalence of LCMV positive urban residents was found to be consistently low (1-3.6%) along the study period, with overage prevalence of 3.3% and values in males (4.6%) significantly higher than in females (2.6%) (P < 0.05). Seven of 432 pregnant women were found to be LCMV positive, but the absence of LCMV antibodies in the newborns demonstrated that the mothers were infected before pregnancy. This study is the first evidence on endemic LCMV in an Argentine city located outside the endemic area of Argentine hemorrhagic fever (AHF) and described the need to study other areas and increase awareness of this viral infection. PMID- 15834872 TI - Human enterovirus isolates from an outbreak typed using heteroduplex mobility analysis. AB - Genotyping and serotyping of enteroviruses is important for epidemiological, prognostic, and therapeutic reasons. In this study clinical isolates of enterovirus 71 during an outbreak of childhood meningoencephalitis in Sydney, Australia were identified using heteroduplex mobility analysis (HMA) of products from RT-PCR amplification of the 5' untranslated region. Five enterovirus 71 isolates shared identical heteroduplex patterns and nucleotide sequences in the 5' untranslated region. A sixth isolate exhibited minor differences in heteroduplex pattern and sequencing confirmed the isolate varied by 1% at the nucleotide level. The use of multiple reference strains and the analysis of heteroduplex patterns increased the confidence of isolate identification, and allowed identification of strain variation which could be subsequently further analyzed using sequencing. HMA can be used to accurately distinguish identical and variant isolates derived from sporadic cases and clustered infections with enteroviruses, including those causing serious infections. PMID- 15834873 TI - A phylogenetic study of human respiratory syncytial viruses group A and B strains isolated in two cities in Japan from 1980-2002. AB - The circulation pattern and genetic evolution of respiratory syncytial virus (RSV) in Japan were examined based on 109 RSV field strains isolated over 20 seasons (1980-2002) in two cities, Sapporo and Tokyo. The second hypervariable region of the large glycoprotein (G) gene was amplified by RT-PCR and the products sequenced directly. The nucleotide sequences were compared to those representatives of RSV genotypes identified previously. Japanese group A and B isolates clustered into five and four genotypes defined previously, respectively. Another one group A and one group B genotypes, which could not be assigned to previous genotypes, were also identified. Although different genotypes usually co circulated in each season, the isolates in proximate seasons from two communities were usually located in the same branches. Moreover, the strains with genotypes defined previously were usually isolated at the same time as each reference strain of Western countries. Several mutant group B strains with 1-20 longer amino acid G proteins were newly identified in Sapporo. These findings suggest that Japanese RSV strains underwent geographical and also temporal clustering while participating in RSV genetic evolution in a global setting. In addition, Japanese strains, especially group B, might have evolved individually in each community, sometimes changing the length of the G protein. PMID- 15834874 TI - Nucleotide mutations associated with hepatitis B e antigen negativity. AB - One hundred and forty four patients with chronic hepatitis B were tested to identify new mutations associated with hepatitis B e antigen (HBeAg) negativity, using a full genome sequence analysis. All the patients were Chinese and had hepatitis B virus infection of genotype C. Patients with none of the pre-core or core promoter mutations were significantly (P < 0.001) less common in the group with anti-HBe (13%) than in the group with HBeAg (56%). The complete nucleotide sequence was determined in four anti-HBe-positive patients who had neither pre core nor core promoter mutations and in five HBeAg-positive patients who also had neither of these mutations (the groups were matched for age and sex). Six mutations were found to be significantly more common in the former group than in the latter: G529A (3/4 vs. 0/5), C934A (4/4 vs. 1/5), A1053G (4/4 vs. 1/5), G1915T/A (4/4 vs. 0/5), T2005C/A (4/4 vs. 0/5), and C3026T (3/4 vs. 0/5). Three of the six mutations were significantly more common in the four anti-HBe-positive patients who had neither pre-core nor core promoter mutations, compared to 11 HBeAg-positive patients who had pre-core and core promoter mutations, and also compared to 15 anti-HBe-positive patients who had pre-core and core promoter mutations, suggesting further the specificity of these mutations. Of the six mutations, two resulted in amino acid substitution in the polymerase protein, and one is located near the enhancer I region. The results suggest that the six newly discovered mutations are associated with HBeAg negativity. PMID- 15834875 TI - Preliminary study on the efficacy and safety of lamivudine and interferon alpha therapy in decreasing serum HBV DNA level in HBV positive transgenic mice during pregnancy. AB - Previous studies reported that the HBV DNA level in maternal serum is an important risk factor for intrauterine infection. Two antiviral drugs, lamivudine (3TC) and interferon alpha (IFNalpha), are used extensively clinically to reduce maternal HBV DNA level, However, because of a lack of evidence on the efficacy and safety of these drugs during pregnancy, they are categorized as grade C which prevents their use during pregnancy. This study provides new data on the efficacy and safety of lamivudine and IFNalpha in HBV positive transgenic pregnant mice. In this study, transgenic mice with high titers of hepatitis B virus (HBV) were employed to study the antiviral effects of 3TC and IFNalpha during different gestation periods. The study also examined changes in several serological HBV markers, the effects of perinatal exposure to antiviral drugs on the mother and offspring, drug efficacy in reducing the level of HBV DNA in maternal blood, and the safety to both the mother and offspring. The main conclusion of the study is that a significant decrease in HBV DNA level can be obtained after treatment with lamivudine but not with IFNalpha. No adverse effects were observed in the maternal mice and the offsprings. This finding may provide a rationale for the potential use of lamivudine for the treatment of pregnant women as a safe and effective measure to reduce the level of maternal viremia. PMID- 15834876 TI - Oligonucleotide array for simultaneous detection of respiratory viruses using a reverse-line blot hybridization assay. AB - The interest in developing new diagnostic methods based on arrays of multiple probes to detect and type simultaneously a wide range of different infectious agents is increasing. This becomes a necessity in the case of infectious agents such as respiratory viruses that cause diseases with very similar signs and symptoms. Such tools will permit rapid and accurate diagnosis of different agents causing respiratory infection leading to the most adequate prevention and/or treatment measures. In this article a reverse-line blot hybridization (RLB) assay for the detection of a wide range of respiratory viruses is presented and evaluated for its usefulness in routine diagnosis. This assay employs an array of 18 oligonucleotide probes immobilized on a nylon membrane. Biotin-labeled PCR products obtained with two multiplex reverse transcription (RT)-polymerase chain reaction (PCR) assays described previously, which allow for the detection of fourteen different groups of respiratory viruses, were hybridized to the oligonucleotide array. Detection was performed using a chemiluminescent method. The standardization of the method showed that the RLB assay could be an alternative to the nested PCR assay for enhancing the sensitivity in the detection of the amplified products, avoiding the problem of cross-over contamination, increasing the specificity, and therefore simplifying the method. This is of main interest in laboratories with few facilities. The feasibility and accuracy of the RT-PCR-RLB assay for detecting respiratory viruses proves that such approach could be a first stage to develop a microarray assay for routine diagnosis of infectious diseases. PMID- 15834877 TI - Dichotomy in cross-clade reactivity and neutralization by HIV-1 sera: Implications for active and passive immunotherapy. AB - The identification of broadly reactive and cross-clade neutralizing antibodies will facilitate the development of a more universally effective vaccine for human immunodeficiency virus (HIV). Antibodies in sera from individuals infected with Clade B HIV bind native primary viral isolates, and virus binding correlates with neutralization and stable clinical disease. In this study, we quantified cross clade antibody reactivity and neutralization by Clades B and C sera. Primary viral isolates were captured by serum IgG bound to anti-human IgG and quantitated as p24 released by lysis of captured virus. Neutralization was determined using PHA-stimulated PBMC. Clade B antibodies reacted more frequently with Clade B R5 virus, but positive sera captured quantitatively more X4 virus than R5 and R5X4 virus. Clade B sera reacted less frequently and captured less Clade C virus than Clade B virus. Antibodies in Clade C sera captured Clades B and C isolates with equal frequency and quantity. There was no difference in neutralization of Clade B virus by either group of sera; however, Clade C sera neutralized Clade C virus, whereas Clade B sera were ineffective against Clade C virus. Thus, there are distinct differences in cross-clade reactivity of and neutralization by antibodies induced in response to Clade C infection compared to Clade B infection. Understanding antibody responses to native virions after Clade C infection and cross clade antibody behavior has implications for understanding pathogenesis and vaccine development. PMID- 15834878 TI - Hepatitis C virus infection in cameroon: A cohort-effect. AB - A hepatitis C virus (HCV) serological study conducted in 2003 on 1,434 individuals in Yaounde and other HCV seroepidemiological studies on 2,066 sera sampled between 1993 and 1997 in four geographically distinct rural areas (Ntem, Mekas, Yokadouma, and Nditam) in Cameroon, are described. Two patterns of HCV seroprevalence were observed. The first pattern, represented by Nditam and Yokadouma populations, showed low HCV seroprevalence rates (2.9% and 3.3%, respectively) increasing moderately with age (9.0% and 16.7% after age 50). The second pattern showed high seroprevalence rates (6.9% for Yaounde, 14.4% and 16.7% for Ntem and Mekas, respectively). These rates increased dramatically with age (32.8%-49.5% after age 50). The age-specific anti-HCV prevalence curve of the 1993 Mekas survey paralleled those of the 1997 Ntem and 2003 Yaounde surveys. Using the year of birth as the x-axis, the three curves closely matched each other. This clearly indicates a cohort effect for which the seroprevalence trends are clearly related with the year of birth, rather than the age. The highest prevalence was observed among people born around 1940. PMID- 15834879 TI - HFRS causing hantaviruses do not induce apoptosis in confluent Vero E6 and A-549 cells. AB - Hantaviruses are known to cause little or no cytopathic effect in vitro, but have been suggested to cause apoptosis. To determine whether different hantaviruses would induce apoptosis to varying degrees, confluent Vero E6 cells were infected with the hemorrhagic fever with renal syndrome (HFRS) causing viruses Hantaan, Dobrava, Saaremaa, and Puumala. However, no difference was found in the percentage of adherent cells, or of cells with condensed nuclei, between non infected and virus-infected cells at 3, 6, 9, or 12 days after infection. Furthermore, no differences in the percentage of cells with inter-nucleosomal cleavage of DNA between uninfected and Hantaan infected cells could be detected using the TUNEL assay. Possibly, slightly more apoptotic cells, but never more than 5%, were detected after Hantaan infection of non-confluent cells as compared to the negative control. Earlier reported results that Tula hantavirus induces significant apoptosis on Vero E6 cells were also verified, suggesting that non pathogenic hantaviruses might differ from HFRS-causing strains regarding induction of apoptosis. In conclusion, the results indicated that the HFRS causing hantaviruses might induce a very low level of apoptosis in dividing cells, but not at all in confluent cells. PMID- 15834880 TI - Human T-cell lymphotropic virus antibody prevalence in HIV-1-infected individuals attending a sexual health clinic in South-East London. AB - Human immunodeficiency virus (HIV) and human T-cell lymphotropic virus (HTLV) are both retroviruses with similar routes of transmission. A number of reports suggest variable clinical outcomes in HIV and HTLV co-infected individuals. There is no published information regarding the prevalence of HIV and HTLV co-infection in the UK. We therefore carried out an unlinked anonymised retrospective study to investigate the prevalence of HTLV co-infection in HIV infected patients attending a sexual health clinic in South-East London. We identified sera from 777 HIV-1 positive adults (504 male, 273 female) who had attended our sexual health clinic between January 2000 and March 2001. Serum samples stored at -20 degrees C were initially tested by HTLV-1 and HTLV-2 antibody enzyme-linked immunoassay (EIA). An immunoblot assay was carried out on reactive samples to discriminate between viral subtypes. Samples with indeterminate results were also analyzed by Western blot. The prevalence of HTLV antibody was 0.8% (five patients with HTLV-1 and one with HTLV-2). Four of the HTLV-1 co-infected patients were females born abroad, of Black African or Caribbean origin. The other HTLV-1 infected patient was a male in the Black Other ethnic group born in the UK, demonstrating that transmission may occur outside recognized areas of high endemicity. The HTLV-2 co-infected individual was a White male born in Italy, and was likely to have been infected through intravenous drug use. The results suggest HTLV antibody screening should be considered in the local HIV infected population of south London. PMID- 15834881 TI - Prevalence of naturally occurring surface antigen variants of hepatitis B virus in Korean patients infected chronically. AB - Although Korea is one of the endemic areas of hepatitis B virus (HBV) infection, the prevalence of naturally occurring variants in the major hydrophilic region (MHR) of the surface (S) gene of HBV has not been determined. In the present study, the prevalence of these variants was examined in terms of the clinical state, and HBeAg serostatus in a large series of Korean patients with chronic HBV infection by direct sequencing analysis of part of the S gene containing the MHR of HBV isolated from 101 chronic HBV patients (51 HBeAg-positive and 50 HBeAg negative): 37 were asymptomatic carriers, 21 had chronic hepatitis, 20 had liver cirrhosis, and 23 had hepatocellular carcinoma (HCC). Forty-seven MHR variants (46.5%) of the 101 patients were detected, involving a total of 59 amino acid substitutions at 12 positions inside and 14 position outside the 'a' determinant, and 33 'a' determinant variants (32.7%). A total of 17 novel variants and 14 novel mutation patterns were detected. The prevalence of MHR variants in HBeAg negative patients tended to be higher than in HBeAg-positive patients (54.0% vs.39.2%) and the prevalence of MHR variants in HCC and liver cirrhosis tended to be higher than in asymptomatic carriers (65.2% vs. 40.5% and 50.0% vs. 40.5%, respectively). In conclusion, three important findings were found in the present study. First, an unexpectedly high prevalence of naturally occurring MHR variants was found in Korean chronic patients. Second, several novel variants associated with mutations outside the 'a' determinant were detected. Finally, a higher prevalence of MHR variants was associated with HBeAg-negative serostatus and severe liver disease, particularly HCC. PMID- 15834882 TI - Viraemia is a common finding in immunocompetent children with rotavirus infection. AB - Rotavirus infection is usually localized to the intestine but involvement of extra-intestinal sites, including the respiratory tract, liver, kidney, lymph nodes, and central nervous system, has been reported. The extra-intestinal spread of the virus may occur through blood since viraemia has been documented occasionally in animals and humans. Nevertheless, the questions of how common viraemia is in immunocompetent children and whether it is associated with extra intestinal manifestations remain unsolved. Serum samples from 54 immunocompetent children hospitalized for acute diarrhea were evaluated prospectively for the presence of rotavirus RNA by nested reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate this issue. Rotavirus antigens were detected in the stools of 14 children. A bacterial aetiology was documented in 14 cases, and in the remaining cases the aetiology remained unknown. Rotavirus RNA was detected in the blood of 9/14 (64.3%) rotavirus infected children but in no child with diarrhea of other origin. Positive RT-PCR was associated with high fever and/or evidence of extra-intestinal involvement. All positive samples were collected within 3 days of illness onset, suggesting that viraemia was detectable for only a few days. Children in whom rotavirus was detected only in stool samples had high fever but no other extra-intestinal feature. These data suggest that viraemia is common in children infected with rotavirus, which may be associated with extra-intestinal involvement. PMID- 15834883 TI - Rapid investigation of hepatitis A virus outbreak by single strand conformation polymorphism analysis. AB - Investigation of hepatitis A virus (HAV) outbreaks often implies nucleotide sequence analysis. As an alternative method for the identification of related strains, single strand conformation polymorphism method (SSCP) was compared to sequence analysis. Twenty-three strains from sporadic and outbreak cases were studied retrospectively. SSCP, sequence identity and phylogenetic analyses were conducted on a 267 bp fragment of the VP1-2A variable region. The results of SSCP pattern comparison and sequence identity were highly correlated (r = 0.92, P < 0.001). If SSCP showed similar patterns, the VP1-2A fragments had a high and significant probability to have a sequence identity over 99.6%. Results were concordant for outbreak strains. The only discordant result concerned a cluster of three sporadic cases evidenced by phylogenetic analysis while SSCP showed similar patterns for only two of these three cases. A prospective SSCP analysis of a recent HAV outbreak confirmed the reliability of this technique. SSCP may thus provide a rapid and cost-effective tool for preliminary investigation of HAV outbreaks, before undertaking exhaustive nucleotide sequence analysis. PMID- 15834884 TI - Contrasting patterns of hepatitis C virus infection in two regions from Tunisia. AB - This report is a population-based study describing the pattern of hepatitis C virus (HCV) infection in two distinct regions in Tunisia. The study included a total of 11,507 individuals sampled in 1996 from both genders, all age groups, urban and rural settings belonging to 2,973 families. HCV infection was assessed by commercial enzyme immunoassay (EIA) and immunoblot assays and detection of HCV RNA by PCR. HCV genotypes and subtypes were determined by sequencing in the 5' untranslated region (UTR) viral genomic region and the INNO-LiPA HCVII genotyping kit. Genetic relatedness between HCV strains was assessed by sequencing of a portion of the NS5B region. HCV prevalence was significantly higher in the North Western region than in the Southern one: 1.7% versus 0.2% (P < 10(-3), chi(2) = 8,506). There was no difference in positivity according to gender or living in rural or urban settings; the only significant risk factor was advanced age. HCV prevalence among household contacts of HCV positives was not significantly higher than the prevalence in the whole study population. These results indicate a heterogeneity in the geographical distribution of HCV in Tunisia. An increased HCV transmission occurs in the North-Western region with large predominance of genotype 1b (88%) and low contribution of intrafamilial transmission. PMID- 15834885 TI - Estimates of global research productivity in virology. AB - The quantity and quality of published research in the field of Virology by different world regions was estimated in this study. Using the PubMed database, articles from journals included in the "Virology" category of the "Journal Citation Reports" database of the Institute for Scientific Information for the period 1995-2003 were retrieved. The world was divided into nine regions based on geographic, economic, and scientific criteria. Data on the country of origin of the research was available for 33,425 out of 33,712 articles (99.2% of all articles from the included journals). USA exceeds all other world regions in research production for the period studied (42% of total articles), with Western Europe ranking second (35.7%). The mean impact factor in articles published in Virology journals was highest for the USA (4.60), while it was 3.90 for Western Europe and 3.22 for the rest of the world (seven regions combined). USA and Canada ranked first in research productivity when both gross national income per capita (GNIPC) and population were taken into account. The results of this analysis show a distressing fact; the absolute and relative production of research in the field of Virology by the developing regions is very low, although viral diseases cause considerable morbidity and mortality in these areas. It is evident from this study that developing regions need more help from the developed regions to enhance research infrastructure. PMID- 15834886 TI - Fruit sharing between wild adult chimpanzees (Pan troglodytes schweinfurthii): a socially significant event? AB - Although food sharing is a habitual aspect of chimpanzee (Pan troglodytes) life, sharing of plant foods between unrelated adults is rare. Observations of such behavior have typically been interpreted as the outcome of a process by which individuals that are otherwise unable to gain access to the food manage to obtain a nutritional benefit. Here we present behavioral details and an acoustic analysis regarding an observation of food sharing between unrelated adult wild chimpanzees that we suggest cannot be explained using traditional nutrition-based models. Instead we propose that the exchange is only understandable as a socially important event, and we cite two further observations in the same population that support this suggestion. PMID- 15834887 TI - Distribution of genotype/subtype and mutational spectra of the surface gene of hepatitis B virus circulating in Hanoi, Vietnam. AB - In order to ascertain the molecular epidemiological features and mutational spectra of hepatitis B virus (HBV) in Hanoi, Vietnam, direct sequencing of the 219-nucleotide fragment of the surface (S) gene of HBV from the sera of 40 patients mostly with chronic hepatitis were carried out. The samples were classified into genotypes by phylogenetic and genotype-specific analysis, and subtypes by the deduced amino acid sequences. The results showed that genotype B with ayw1 was predominant genotype/subtype (63%), followed by genotype C with adr (18%). The quasi-species nature of the HBV in the sera was observed in 24 of 40 samples examined. One sample (HN109) showed mixture of genotypes B and C. Among 26 amino acid substitutions, 16 were the variants and the remainders were mutations. In the "a" determinant region, three mutations with methionine to leucine (L) changes at the 133 amino acid residue were in the first loop and no mutations were in the second loop. A new mutation, threonine to methionine at 126 amino acid residue, was observed in one sample. In conclusion, the analysis of the S gene region of HBV showed that in Hanoi, genotype B with ayw1 was prevalent and the quasi-species nature of HBV was also common. PMID- 15834888 TI - Social interaction with non-averse group-mates modifies a learned food aversion in single- and mixed-species groups of tamarins (Saguinus fuscicollis and S. labiatus). AB - For social species, being a member of a cohesive group and performing activities as a coordinated unit appear to provide a mechanism for the efficient transmission of information about food. Social learning about food palatability was investigated in two captive primates, Saguinus fuscicollis and S. labiatus, which form stable and cohesive mixed-species groups in the wild. We explored whether an induced food aversion toward a preferred food is modified during and after social interaction with non-averse conspecifics or congeners. Sets of intra and interspecific pairs were presented with two foods, one of which was considered distasteful by one of the pairs (the other was palatable), and their behavior was compared pre-interaction, during interaction, and post-interaction. For the aversely-conditioned individuals of both species, the change in social context corresponded to a change in their preference for the food that they considered unpalatable, regardless of whether they had interacted with a conspecific or congeneric pair, and the change in food preference was maintained post-interaction. In a control condition, in which averse individuals did not have the opportunity to interact with non-averse animals, S. fuscicollis sampled the preferred food, but not as quickly as when given the opportunity to interact. We conclude that the social learning demonstrated here may allow individual tamarins to track environmental change, such as fruit ripening, more efficiently than asocial learning alone, because social learners can more quickly and safely focus on appropriate behavior by sharing up-to-date foraging information. Furthermore, since the behavior of congeners, as well as conspecifics, acts to influence food choice in a more adaptive direction, social learning about food palatability may be an advantage of mixed-species group formation to tamarins of both species. PMID- 15834889 TI - Developmental changes in the facial morphology of the Borneo orangutan (Pongo pygmaeus): possible signals in visual communication. AB - Orangutans display remarkable developmental changes and sexual differences in facial morphology, such as the flanges or cheek-pads that develop only on the face of dominant adult males. These changes suggest that facial morphology is an important factor in visual communication. However, developmental changes in facial morphology have not been examined in detail. We studied developmental changes in the facial morphology of the Borneo orangutan (Pongo pygmaeus) by observing 79 individuals of various ages living in the Sepilok Orangutan Rehabilitation Centre (SORC) in Malaysia and in Japanese zoos. We also analyzed photographs of one captive male that were taken over a period of more than 16 years. There were clear morphological changes that occurred with growth, and we identified previously unreported sexual and developmental differences in facial morphology. Light-colored skin around the eyes and mouth is most prominent in animals younger than 3 years, and rapidly decreases in area through the age of approximately 7 years. At the same time, the scattered, erect hairs on the head (infant hair) become thick, dense hairs lying on the head (adult hair) in both sexes. The results suggest that these features are infant signals, and that adult signals may include darkened face color, adult hair, whiskers, and a beard, which begin to develop after the age of approximately 7 years in both sexes. In females, the eyelids remain white even after 10 years, and turn black at around the age of 20; in males, the eyelids turn black before the age of 10. The whiskers and beards of adults are thicker in males than in females, and are fully developed before the age of 10 in males, while they begin to develop in females only after approximately 20 years. White eyelids and undeveloped whiskers and beards may be visual signals that are indicative of young adult females. Our results also show that the facial morphology of the unflanged male is similar to that of the adult female, although it has also been pointed out that unflanged males resemble younger individuals. PMID- 15834890 TI - Social influences on the acquisition of sex-typical foraging patterns by juveniles in a group of wild tufted capuchin monkeys (Cebus nigritus). AB - Foraging traditions in primates are becoming the subject of increasing debate. Recent evidence for such a phenomenon was recently provided for wild Cebus capucinus [Fragaszy & Perry, 2003]. To better understand the bases of animal traditions, one should examine intrapopulation behavioral variability and the influence of social context on within-group transmission of specific foraging patterns. We studied the variability of foraging patterns across age and sex classes, and the proximity patterns of juveniles to adults of both sexes in a group of wild tufted capuchin monkeys (Cebus nigritus) living in the Iguazu National Park, Argentina. Foraging activity was examined for a period of 9 months in terms of proportions of focal samples devoted to foraging on certain food targets, microhabitats, and supports, and using specific foraging patterns. Proximity analyses were performed to reveal patterns of association between juveniles and adults. Sex differences in foraging behavior were present and overrode age differences. Overall, males ate more animal foods, foraged more for invertebrates on woody microhabitats (especially large branches), palms, and epiphytes, and used lower and larger supports than females. Females ate more fruits, foraged more on leaves and bamboo microhabitats, and used smaller supports than males. Juveniles were similar to adults of the same sex in terms of food targets, foraging substrates, and choice of supports, but were less efficient than adults. Proximity patterns indicated that juvenile males stayed in close spatial association with adult males and preferentially focused their "food interest" on them. This phenomenon was less evident in juvenile females. The degree to which juveniles, especially males, showed some of the sex-typical foraging patterns correlated positively with their proximity to adults of the same sex. These findings suggest that the acquisition of foraging behaviors by juvenile males is socially biased by their closeness to adults of the same sex. PMID- 15834891 TI - Tool use in insect foraging by the chimpanzees of Ngogo, Kibale National Park, Uganda. AB - Chimpanzee tool use for resource acquisition has been reported at numerous research sites. The chimpanzees of the Kibale Forest, western Uganda, have not previously been observed to use tools in foraging for insects. Here I report the first observation of tool use by the chimpanzees of the Ngogo community of Kibale National Park, Uganda, in insect foraging. Three adult females, one adolescent male, and one juvenile male were observed making and using tools to probe into a fallen dead tree to collect insect and wood pieces. I discuss the importance of this observation, and the behavioral similarities with chimpanzees from other sites. PMID- 15834892 TI - Adult-infant food transfer in common marmosets: an experimental study. AB - Infant marmosets and tamarins obtain solid food items from adults during and after the time of weaning. In addition to providing nutrients, food transfers may provide infants with the opportunity to learn about diet. The aim of this study was to investigate patterns of begging and food transfer in captive groups of common marmosets during tests with 1) palatable novel food, 2) unpalatable novel food, and 3) familiar food. Infants may gain an opportunity to learn about diet breadth by preferentially begging for novel rather than familiar items, while adults may facilitate infant learning by differing in their willingness to transfer food depending upon the food's novelty or palatability. In this study, infants exhibited more interest and begging with novel food items than with familiar ones. However, food transfers were not more likely to occur with novel foods than with familiar ones, and in fact adults were more likely to refuse infant begging attempts with novel foods. The palatability of the food also did not influence the rates of refusal and transfer: adults transferred palatable and unpalatable novel foods with similar frequencies. These results suggest that infant marmosets gain an opportunity to learn about diet breadth by begging for novel foods, but adults do not preferentially transfer novel or palatable food items. PMID- 15834893 TI - Female sooty mangabeys (Cercocebus torquatus atys) respond differently to males depending on the male's residence status-preliminary data. AB - I tested the hypothesis that free-ranging female mangabeys (Cercocebus torquatus atys) differentiate between males depending on their residence status. Adult males in this species employ two group-membership strategies: they are either full-time residents or part-time residents (i.e., present for several weeks at a time, then gone for several weeks, then present again, etc.). In a playback experiment I tested whether adult females with young infants responded differently to calls of adult males that were group members during the previous mating season as compared to males that belonged to neighboring groups during the mating season or were complete strangers. Males whose calls were played were present residents, part-time residents (absent for at least 3 months), neighbors, or complete strangers. Because infanticide is a potential risk in mangabeys, I predicted that females would respond differently to males that had resided in the group when their infant was conceived than they would to those that had not been present. The females showed a clear difference in their reaction to the playbacks of vocalizations from resident and part-time resident males as opposed to nonresidents. PMID- 15834895 TI - 50-Hz magnetic field (0.1-mT) alters c-fos mRNA expression of early post implantation mouse embryos and serum estradiol levels of gravid mice. AB - BACKGROUND: We exposed pregnant mice to magnetic or sham fields for 24 hr/day during Gestation Day (GD) 5.5-8.5 to study the effects of 50-Hz 0.1-mT sinusoidal magnetic fields on early pregnancy in mouse embryos and mice. METHODS: Mice were sacrificed on GD 8.5. Embryos and blood samples were collected. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of embryo c-fos mRNA. The blood samples were tested through radioimmunoassay for serum estradiol levels and general examinations. RESULTS: We found that the expression of c-fos mRNA of embryos under exposure was enhanced. We reported that 50-Hz 0.1-mT magnetic exposure induced the decline of serum estradiol levels of pregnancy mice on GD 8.5. General examinations of blood including white blood cell (WBC) count, red blood cell (RBC) count, and hemoglobin (HGB) did not show significant differences between the exposure group and the control group (sham exposure). CONCLUSIONS: Under the experimental conditions, 50-Hz 0.1-mT sinusoidal magnetic fields affected the development of early pregnancy mouse embryos and the dams to some extent. PMID- 15834894 TI - Developmental toxicity research of ginsenoside Rb1 using a whole mouse embryo culture model. AB - BACKGROUND: Ginseng has been widely used around the world for many years. Knowledge is limited, however, on its effects on embryonic development. METHODS: Whole embryo culture was used to explore the developmental toxicity of ginsenoside Rb1 (GRb1) on mouse embryos. All embryos were exposed to different concentrations of GRb1, and scored for their growth and differentiation at the end of the 48-hr culture period. RESULTS: Total morphological score decreased significantly at the concentration of GRb1 of 30 microg/ml and was further reduced at 50 microg/ml. Yolk sac was affected at the lower concentration of 15 microg/ml. Developments of midbrain, forebrain, and optic system were relatively sensitive to GRb1 and were affected at the concentration of 30 microg/ml. Allantois, flexion, branchial arch, and limb buds were affected at 50 microg/ml. At this concentration, the embryonic crown-rump length, head length, and somite number were also reduced significantly compared to the control group. CONCLUSIONS: These results suggest that GRb1 has teratogenic effect during the mouse organogenetic period. We suggest that before more data in humans is available, ginseng should be used with caution by pregnant women in the first trimester. PMID- 15834897 TI - Maternal antioxidant supplementation does not reduce the incidence of phenytoin induced cleft lip and related malformations in rats. AB - There is considerable evidence that phenytoin-induced birth defects in the rat are a consequence of a period of bradycardia and hypoxia in the embryos. Experiments were designed to test the hypothesis that phenytoin-induced birth defects result from free-radical damage to the embryos during the reoxygenation period posthypoxia. Female rats (>9 per group) were fed either a control diet or a diet high in antioxidants (vitamins C and E and coenzyme Q(10)) both before and during pregnancy and were then given a teratogenic dose of phenytoin (180 mg/kg) on GD 11. The rats were killed on GD 20 and the fetuses were examined for malformations. The initial results showed that the antioxidant diet had a significant protective effect, with far fewer antioxidant-group fetuses showing cleft lip or maxillary hypoplasia compared with the control group. However, this result was confounded by reduced food intake by the rats fed the antioxidant diet and a significantly lower maternal body weight at the time of phenytoin administration. Since the phenytoin was administered by intraperitoneal injection (i.p.) the control rats received higher absolute doses of phenytoin and it is speculated that this results in higher fetal exposure. A second experiment, in which the rats were pair-fed, failed to demonstrate any protective effect of the high antioxidant diet. These results do not support the reoxygenation hypothesis for phenytoin teratogenesis. An alternative explanation would be hypoxia-induced transcription-related changes resulting in cell cycle arrest and apoptosis. PMID- 15834898 TI - Evaluation of the gene expression changes induced by 17-alpha-ethynyl estradiol in the immature uterus/ovaries of the rat using high density oligonucleotide arrays. AB - BACKGROUND: In a previous study, we determined the effects of 17-alpha-ethynyl estradiol (EE) on gene expression using microarrays that represented approximately 9,000 genes, which was the state of-the-art. Higher content arrays with almost double the number of genes have since become available. In order to better determine whether common sets of gene expression changes can be predictive of estrogenic activity, we have replicated the previous experiment using the more comprehensive microarray. METHODS: Immature 20-day-old Sprague-Dawley rats were exposed to 0.1, 1, and 10 microg EE/kg/day ( subcutaneously [s.c.]), for four days, dosing from postnatal day (PND) 20-23). Changes in a more comprehensive expression level of 15,923 rat annotated genes and expressed sequence tags were evaluated on PND 24. RESULTS: By comparing the response of the treatment groups versus controls using various statistical parameters, we determined that the expression of 1,394 genes showed a significant change with respect to control (p< or =0.0001), to at least one of the EE dosages. The tissues from animals exposed to 0.1 microg EE/kg/day showed changes in the expression of only 33 genes, whereas when they were exposed to 1 or 10 microg EE/kg/day, the expression of 409 and 548 genes was modified, respectively. A dose-dependent analysis indicated that 592 genes showed a robust and significant response to EE exposure (increased or decreased). Our analysis confirmed the regulation of previously identified estrogen-sensitive genes, and clearly identified novel mediators of estrogen actions, both in the uterus as well as in the ovary. CONCLUSIONS: This compendium of genes represents the largest compilation of estrogen-responsive genes that has ever been identified for the immature uterus and ovary of any species, and can be used to generate testable hypothesis to improve the understanding of the molecular pathways associated with physiological and pathophysiological responses to exposure to chemicals with estrogenic properties. PMID- 15834899 TI - Occupational exposures and reproductive health: 2003 Teratology Society Meeting Symposium summary. AB - Assuring reproductive health in the workplace challenges researchers, occupational safety and health practitioners, and clinicians. Most chemicals in the workplace have not been evaluated for reproductive toxicity. Although occupational exposure limits are established to protect 'nearly all' workers, there is little research that characterizes reproductive hazards. For researchers, improvements in epidemiologic design and exposure assessment methods are needed to conduct adequate reproductive studies. Occupational safety and health programs' qualitative and quantitative evaluations of the workplace for reproductive hazards may differ from standardized approaches used for other occupational hazards in that estimates of exposure intensity must be considered in the context of the time-dependent windows of reproductive susceptibility. Clinicians and counselors should place the risk estimate into context by emphasizing the limitations of the available knowledge and the qualitative nature of the exposure estimates, as well as what is known about other non-occupational risk factors for adverse outcomes. This will allow informed decision-making about the need for added protections or alternative duty assignment when a hazard cannot be eliminated. These policies should preserve a worker's income, benefits, and seniority. Applying hazard control technologies and hazard communication training can minimize a worker's risk. Chemical reproductive hazard training is required for workers by the Occupational Safety and Health Administration's Hazard Communication Standard. The National Institute for Occupational Safety and Health (NIOSH) has formed a National Occupational Research Agenda Team to promote communication and partnering among reproductive toxicologists, clinicians and epidemiologists, to improve reproductive hazard exposure assessment and management, and to encourage needed research. PMID- 15834900 TI - Craniofacial and axial skeletal defects induced by the fungicide triadimefon in the mouse. AB - BACKGROUND: Triadimefon is an antifungal derived from triazole. In in vitro whole rodent embryo cultures, triazole-derivatives showed specific teratogenic effects at the branchial apparatus. The aim of the present work was to test in vivo triadimefon (FON), in order to verify a relationship between triazole exposure, embryonic abnormalities, and/or fetal malformations. METHODS: Pregnant CD-1 mice were treated with 0-300 mg/kg FON by gavage on day 8 post coitum (p.c.) at 10:00 AM, and sacrificed on day 8 p.c. at 1:00 PM, on day 9 p.c. at 10:00 AM, on day 10 p.c. at 10:00 AM, and at term of gestation (day 18 p.c.). At midgestation, the embryos were processed for specific immunostainings to visualize the hindbrain segmentation (day 8 p.c.) and the neural crest cell migration (days 8 and 9 p.c.). Fetuses explanted at term were all processed for skeletal examination after double-staining of osseous and cartilaginous tissues. RESULTS: At midgestation, the immunostaining of rhombomeres 3 and 5 showed a light scattering of the immunostained areas; the neural crest cell migration was unaffected, but their localization at the branchial arch level was abnormal. At term, several severe malformations were observed at the craniofacial and at the axial skeletal level. Ectopic cartilage was observed at the upper jaw. CONCLUSIONS: Triadimefon is teratogenic. The observed craniofacial malformations could be explained by an alteration of the rhombomeric organization and neural crest migration to the branchial arches; the axial abnormalities could be explained by the abnormal segmental identity specification. PMID- 15834901 TI - The transport of chemicals in semen. AB - Three mechanisms have been proposed for exposure of the conceptus to chemicals in semen: access of chemicals to the maternal circulation after absorption from the vagina, direct chemical exposure of the conceptus following transport from the vagina to the uterine cavity, and delivery to the egg and subsequent conceptus of chemical bound to the sperm cell. We review published data for each of these three mechanisms. Human seminal fluid chemical concentrations are typically similar to or lower than blood concentrations, although some antimicrobial agents achieve higher concentrations in semen than in blood. Vaginal absorption of medications has been shown to occur, although the vehicles in which these medications are delivered to the vagina may maintain contact with the vaginal epithelium to a greater extent than does semen. Assuming total absorption of a seminal dose of a chemical with a high semen:blood concentration ratio, distribution within the recipient woman would result in a blood concentration at least three orders of magnitude lower than that in the man. Direct delivery of seminal chemicals into the uterine cavity of humans has not been shown to occur, although it may occur in species such as the rat in which seminal fluid has access to the uterine cavity. Chemicals in or on human sperm cells have been demonstrated with respect to tetracycline and cocaine in vitro and aluminum, lead, and cadmium in vivo. The in vitro cocaine study offers sufficiently quantitative data with which to predict that oocyte concentrations would be five orders of magnitude lower than blood concentrations associated with cocaine abuse, assuming a maximally cocaine-bound sperm were capable of fertilizing. Thus, even using liberal assumptions about transmission of chemicals in semen or sperm, predicted exposure levels of a pregnant woman or of the conceptus are three or more orders of magnitude lower than blood concentrations in the man whose semen is the putative vehicle for chemical transport. PMID- 15834902 TI - Postnatal development of the gastrointestinal system: a species comparison. PMID- 15834903 TI - Effect of inversion time on delayed-enhancement magnetic resonance imaging with and without phase-sensitive reconstruction. AB - PURPOSE: To evaluate the consistency and inversion time (TI) independence of phase-sensitive reconstruction (PSIR) delayed-enhancement (DE) MRI in a clinical setting. MATERIALS AND METHODS: Mid-ventricular short-axis DE images were acquired in 25 patients using three TIs: 1) optimized to null viable myocardium, 2) 50 msec less than optimal TI, and 3) 50 msec greater than optimal TI. At each TI, images were acquired with PSIR and without magIR. In each image, percent scar was computed as the ratio of nonviable to total pixels in the left ventricle (LV). RESULTS: In the magIR images, percent scar was 23% +/- 15% (optimal), 11% +/- 11% (short), and 22% +/- 15% (long). In PSIR images, percent scar was 25% +/- 15% (optimal), 22% +/- 15% (short), and 22% +/- 14% (long). Percent scar was significantly underestimated in magIR images with short TI, but no statistically significant difference in percent scar was observed at the optimal or long TIs. CONCLUSION: DE-MRI is a robust imaging technique for clinical use. PSIR provided consistent image quality independently of TI, at least over the range of TIs evaluated in this study. However, neither image quality nor scar appearance in the PSIR images was significantly different from that in the magIR images when TI was at or above the null point of viable myocardium. PMID- 15834904 TI - Subtraction of in-phase and opposed-phase images in dynamic MR mammography. AB - PURPOSE: To develop and to evaluate an advanced image acquisition and analysis method for collecting T(1)-weighted dynamic 3D MR mammography data sets by using a combined in-phase (IP) and opposed-phase (OP) imaging procedure. MATERIALS AND METHODS: 3D MR mammography data sets were acquired by applying an interleaved gradient-echo OP and IP imaging sequence during administration of contrast agent. A phantom data set, two volunteer breast data sets, and six patient breast data sets were recorded. Subtraction of dynamic in-phase magnitude images was performed for clinical assessment. In addition, the magnitude subtraction (SIPOP) as well as the complex subtraction (cSIPOP) of the IP and OP magnitude and phase images were considered. RESULTS: The detection of small lesions, lesion boundaries, and tumor offshoots in fatty tissue was improved by the subtraction of IP and OP images without the risk of signal cancellation due to partial volume effects. CONCLUSION: Dynamic MR mammography acquisition of IP and OP images in combination with appropriate data processing yields important supplementary information that can support routinely applied diagnostics of breast lesions that are fully embedded in fatty tissue by only marginally increasing acquisition time. PMID- 15834905 TI - In vitro validation of phase-contrast flow measurements at 3 T in comparison to 1.5 T: precision, accuracy, and signal-to-noise ratios. AB - PURPOSE: To evaluate the signal-to-noise ratio (SNR), precision, and accuracy of phase-contrast flow measurements at 3 T with the help of an in vitro model and to compare the results with data from two 1.5-T scanners. MATERIALS AND METHODS: Using an identical setup of a laminar flow model and sequence parameters, measurements were done at one 3-T and at two 1.5-T systems. Precision, accuracy, and SNR were obtained for velocity encodings ranging from 55 up to 550 cm(-1). SNRs were calculated from the magnitude as well as the flow encoded images. RESULTS: Precision and accuracy for the in vitro flow model were similarly high in all scanners with no significant difference. For velocity encodings from 55 cm(-1) up to 550 cm(-1), the SNR in magnitude as well as phase encoded images of the 3-T measurements was approximately 2.5 times higher than the SNR obtained from the two 1.5-T systems. CONCLUSION: Even without optimization for the 3-T environment, flow measurements show the same high accuracy and precision as is known from clinical 1.5-T scanners. The superior SNR at 3 T will allow further improvements in temporal and spatial resolution. This will be of interest for small-size vessels like coronary arteries or for slow diastolic flow patterns. PMID- 15834906 TI - MR findings in a rare case of sclerosing mesenteritis of the mesocolon. AB - Sclerosing mesenteritis is a rare, usually benign disorder of the mesentery. Depending on the predominant tissue component (inflammation, fat, or fibrosis), it is known as mesenteric panniculitis or retractile mesenteritis. We present a rare case of retractile mesenteritis of the mesocolon as a cause of severe abdominal pain. US, CT, and MRI were the imaging modalities used. We emphasize the MR finding of a fibrous capsula in retractile mesenteritis, as this is to our knowledge the first study to describe this entity. This finding may be valuable for establishing a diagnosis of sclerosing mesenteritis, as well as for differentiating this disease from other mesenteric diseases. PMID- 15834907 TI - Frequency of malignancy in lesions classified as probably benign after dynamic contrast-enhanced breast MRI examination. AB - PURPOSE: To determine the chance of malignancy in lesions classified as "probably benign" by dynamic magnetic resonance imaging (MRI), in a heterogeneous population. MATERIALS AND METHODS: Reports from 473 patients, from March 1994 to March 2002, who underwent breast MRI were retrospectively reviewed. A total of 79 patients (17%) had lesions classified as probably benign after the MRI, which required further imaging follow-up. We evaluated subsequent MRI, mammographic reports, and clinical follow-up in these patients and established the frequency of malignancy in this group. RESULTS: MRI classified probably benign lesion were diagnosed in 79 women because of focal or diffuse mild enhancement and benign dynamic enhancement curves in the area of the mammographic abnormality, or because of the presence of microcalcifications on the mammogram, or because of incidental enhancing lesions. Two-year radiographic and/or clinical follow-up was available in 68 women. On follow-up, four women (6%) were diagnosed with cancer between 14 and 18 months after the initial MRI. CONCLUSION: Patients with a lesion assessed as probably benign by dynamic contrast enhanced MRI have a higher chance of malignancy than patients with probably benign lesions (Breast Imaging Reporting and Data System category 3, BI-RADS 3) seen on mammography. These patients should be informed of the increased risk of cancer and be given the option of biopsy or close follow-up. PMID- 15834908 TI - Magnetic resonance imaging of the upper abdomen using a free-breathing T2 weighted turbo spin echo sequence with navigator triggered prospective acquisition correction. AB - PURPOSE: To evaluate a free-breathing navigator triggered T2-weighted turbo spin echo sequence with prospective acquisition correction (T2w-PACE-TSE) for MRI of the upper abdomen in comparison to a conventional T2-weighted TSE (T2w-CTSE), a single-shot TSE (T2w-HASTE), and a T1-weighted gradient-echo sequence (T1w FLASH). MATERIALS AND METHODS: A total of 40 consecutive patients were examined at 1.5 T using free-breathing T2w-PACE-TSE, free-breathing T2w-CTSE, and breath hold T2w-HASTE and T1w-FLASH acquisition. Images were evaluated qualitatively by three radiologists regarding motion artifacts, liver-spleen contrast, depiction of intrahepatic vessels, the pancreas and the adrenal glands, and overall image quality on a four-point scale. Quantitative analysis of the liver-spleen contrast was performed. RESULTS: Depiction and sharpness of intrahepatic vessels were rated significantly better (P < 0.01) using T2w-PACE-TSE compared to T2w-CTSE and T2w-HASTE sequences. Significantly higher contrast values were measured for T2w PACE-TSE images compared to T2w-CTSE, T2w-HASTE, and T1w-FLASH images (P < 0.01). Mean examination time of the T2w-PACE-TSE was 7.91 minutes, acquisition time of the T2w-CTSE sequence was 4.52 minutes. CONCLUSION: Prospective acquisition correction is an efficient method for reducing respiratory movement artifacts in T2w-TSE imaging of the upper abdomen. Compared to T2w-CTSE and T2w-HASTE sequences recognition of anatomical details and contrast can be significantly improved. PMID- 15834909 TI - Optimization of a blood pool contrast agent injection protocol for MR angiography. AB - PURPOSE: To design an ideal first-pass profile for MR angiography (MRA) by optimizing a multiphasic injection protocol based on two experimental animal models. MATERIALS AND METHODS: An equivalent contrast-enhanced (CE) MRA injection protocol was developed with controlled injection modalities (injection rate, volume, and dose) in rabbits and pigs. P792, a blood pool contrast agent, was injected in 17 male New Zealand rabbits and five farm pigs with variable injection schemes (mono- and multiphasic). From the gadolinium (Gd) blood concentration data, a simulation of an MR acquisition was performed to evaluate the impact of such an injection protocol on MR arterial signal and to select the best injection protocol. RESULTS: An empirical relationship between the arterial peak concentration and the injection parameters was found in the rabbits and pigs, allowing precise prediction of the first-pass profile. Of the four injection scheme strategies tested (standard bolus and bi-, tri-, and multiphasic injection protocols), the multiphasic "ramp" injection protocol provided the most optimal contrast agent pharmacokinetics with a durable plateau of concentration. CONCLUSION: Ramp injection protocol provides an optimized first-pass profile for CE-MRA. PMID- 15834910 TI - Method for improving the accuracy of quantitative cerebral perfusion imaging. AB - PURPOSE: To improve the accuracy of dynamic susceptibility contrast (DSC) measurements of cerebral blood flow (CBF) and volume (CBV). MATERIALS AND METHODS: In eight volunteers, steady-state CBV (CBV(SS)) was measured using TrueFISP readout of inversion recovery (IR) before and after injection of a bolus of contrast. A standard DSC (STD) perfusion measurement was performed by echo planar imaging (EPI) during passage of the bolus and subsequently used to calculate the CBF (CBF(DSC)) and CBV (CBV(DSC)). The ratio of CBV(SS) to CBV(DSC) was used to calibrate measurements of CBV and CBF on a subject-by-subject basis. RESULTS: Agreement of values of CBV (1.77 +/- 0.27 mL/100 g in white matter (WM), 3.65 +/- 1.04 mL/100 g in gray matter (GM)), and CBF (23.6 +/- 2.4 mL/(100 g min) in WM, 57.3 +/- 18.2 mL/(100 g min) in GM) with published gold-standard values shows improvement after calibration. An F-test comparison of the coefficients of variation of the CBV and CBF showed a significant reduction, with calibration, of the variability of CBV in WM (P < 0.001) and GM (P < 0.03), and of CBF in WM (P < 0.0001). CONCLUSION: The addition of a CBV(SS) measurement to an STD measurement of cerebral perfusion improves the accuracy of CBV and CBF measurements. The method may prove useful for assessing patients suffering from acute stroke. PMID- 15834911 TI - Combined use of the intravascular blood-pool agent, gadomer, and carbon dioxide: a novel type of double-contrast magnetic resonance angiography (MRA). AB - PURPOSE: To evaluate the combined use of carbon dioxide (CO(2)) and a gadolinium based blood-pool agent for magnetic resonance angiography (MRA). MATERIALS AND METHODS: After an initial intravenous injection of the blood-pool agent Gadomer (Schering AG, Germany), repeated transcatheter CO(2) injections were performed in the aorta and the renal arteries of two fully-anesthetized pigs. Real-time images were acquired using a true fast imaging with steady-state precession (FISP) sequence. RESULTS: During the CO(2) injections, the Gadomer-enhanced blood was totally replaced, resulting in an immediate, temporary, total signal loss in the vessel lumen. Susceptibility artifacts during the injections or catheter manipulations rarely occurred. CONCLUSION: Due to T1-shortening, the circulating blood-pool agent prevents flow artifacts during catheter manipulations because the steady-state is reached much earlier. Therefore, this double-contrast MRA method improves catheter conspicuity and might be helpful for guiding and controlling intravascular procedures during interventional MRI. PMID- 15834912 TI - Measuring human cardiac tissue sodium concentrations using surface coils, adiabatic excitation, and twisted projection imaging with minimal T2 losses. AB - PURPOSE: To measure tissue sodium concentrations in the human heart with (23)Na MRI using a surface coil, thereby eliminating the effects of inhomogeneous excitation by surface coils and minimizing T(1) and T(2) relaxation. MATERIALS AND METHODS: We combined fully relaxed, very short-echo, (23)Na twisted projection imaging (TPI) with adiabatic half passage (AHP) excitation and external referencing on subjects and comparing with a concentration reference phantom scan to quantify TSC with surface coils. (23)Na signal losses during hard (square), composite, and tanh/tan amplitude/frequency-modulated AHP excitation pulses were analyzed over a wide range of RF field strengths and T(2short) values. RESULTS: AHP excitation yielded a homogeneous excitation flip angle and negligible losses compared to a 90 degrees hard pulse wherever the B1 field exceeded the adiabatic threshold, rendering this sequence suitable for applications that use surface coil excitation. An AHP (23)Na TPI sequence was used with a surface coil at 1.5 T to noninvasively quantify myocardial TSC in 10 normal volunteers. The mean TSC was 43 +/- 4, 53 +/- 12, and 17 +/- 4 micromol/g in the left ventricular (LV) free wall, septum, and adipose tissue, respectively, consistent with prior invasive measurements on biopsy and autopsy specimens. CONCLUSION: It is now possible to noninvasively quantify TSC in the human heart with surface coil (23)Na MRI. PMID- 15834913 TI - Multisite trial of MR flow measurement: phantom and protocol design. AB - PURPOSE: To describe a portable, easily assembled phantom with well-defined bore geometry together with a series of tests that will form the basis of a standardized quality assurance protocol in a multicenter trial of flow measurement by the MR phase mapping technique. MATERIALS AND METHODS: The phantom consists of silicone polymer layers containing parallel straight and stenosed flow channels in one layer and a U-bend in a second layer, separated by hermetically sealed agarose slabs. The phantom is constructed by casting low melting-point metal in an aluminum mold precisely milled to the desired geometry, and then using the low melting-point metal core as a negative around which the silicone is allowed to set. By melting out the metal, the flow channels are established. The milled aluminum mold is reusable, ensuring faithful reproduction of the flow geometry for all phantoms thus produced. The agarose layers provide additional loading and static background signal for background correction. With the use of the described phantom, one can evaluate flow measurement accuracy and repeatability, as well as the influence of several imaging geometry factors: slice offset, in-plane position, and slice-flow obliquity. RESULTS: The new phantom is compact and portable, and is well suited for reassembly. We were able to demonstrate its facility in a battery of tests of interest in evaluating MR flow measurements. CONCLUSION: The phantom is a robust standardized test object for use in a multicenter trial. Such a trial, to investigate the performance of MR flow measurement using the phantom and the tests we describe, has been initiated. PMID- 15834914 TI - An MRI-compatible semiautomated vacuum-assisted breast biopsy system: initial feasibility study. AB - PURPOSE: To develop an MR-compatible vacuum-assisted core needle breast biopsy system. MATERIALS AND METHODS: A vacuum-assisted breast biopsy system (Mammotome Hand Held; Ethicon Endo-Surgery, USA) was modified for freehand MRI-guided biopsy in an open, interventional 0.5-T scanner (Signa SP; GE, USA). Probes (11 gauge [G]) were fabricated without significant susceptibility artifact. These mate with an electromechanical hand piece and control system that were modified for use within the MRI scanner. A total of 62 breast lesions were simulated in the mammary tissues of six recently lactating sows by injecting between 0.1 and 1.0 mL of an aqueous gel containing dilute gadopentetate dimeglumine (Gd-DTPA) that formed a bright mass on T1-weighted imaging. RESULTS: Mechanical performance was satisfactory. Magnetic susceptibility and radiofrequency (RF) artifacts from the 11-G probe were negligible. T1-weighted fast spin echo (FSE) was used to guide biopsy. Up to eight samples were removed per lesion. Overall, 461 samples were obtained in 493 attempts (94%). Sample weights averaged 54 mg (N = 493) compared to 4.6 mg (N = 24) from 14-G titanium MRI-compatible needles. On average, 59% of the attempted samples yielded target lesion material. CONCLUSION: Preliminary results demonstrate the feasibility of a vacuum-assisted breast biopsy system in the MRI environment. Small 0.1-mL lesions can be biopsied without needle artifacts obscuring the target. PMID- 15834915 TI - An improved method for spinal functional MRI with large volume coverage of the spinal cord. AB - PURPOSE: To develop a spinal functional MRI (fMRI) method with three-dimensional coverage of a large extent of the spinal cord with minimal partial volume effects MATERIALS AND METHODS: fMRI data of the cervical spinal cord were obtained at 1.5 T with a single-shot fast spin-echo imaging method, from thin contiguous sagittal slices spanning the cord. Thermal stimulation was applied to the palm of the hand in a block pattern with 15 degrees C for stimulation and 32 degrees C during baseline periods. Prior to analysis, the image data at each time point were reformatted into three-dimensional volumes and resliced perfectly transverse to the spinal cord. Smoothing was applied only in the superior-inferior (S/I) direction across uniform tissue types. Active voxels were then identified by means of a correlation to a model paradigm. RESULTS: The resulting activity maps demonstrate activity primarily in ipsilateral sensory areas and in some motor areas, consistent with the spinal cord neuroanatomy. These data also demonstrate detail of the subsegmental organization of the spinal cord, as well as anatomical detail of the spinous processes and positions of nerve roots. CONCLUSION: The spinal fMRI method described enables large volume coverage of the spinal cord in three dimensions, with reliable and reproducible results. PMID- 15834916 TI - Gd(ABE-DTTA)-enhanced cardiac MRI for the diagnosis of ischemic events in the heart. AB - PURPOSE: To demonstrate that contrast-enhanced MRI (ceMRI) with the aid of Gd(ABE DTTA) is able to detect ischemic events in the heart in a canine ischemia/reperfusion (30/40 minutes) model. MATERIALS AND METHODS: ECG-gated, T1 weighted MR image sets (four to five slices each) with three-minute time resolution were collected in transiently LAD-occluded dogs. Following the acquisition of control image sets, ischemia was started by occluding the LAD. Either Gd(ABE-DTTA) (N = 6) or Gd(DTPA) (N = 6) was injected, and imaging was continued for 30 minutes of ischemia and 40 minutes of reperfusion. The contrast agent (CA)-induced MRI signal intensity enhancement (SIE) and contrast were monitored. Microspheres measured myocardial perfusion (MP) to verify areas of ischemia and reperfusion. RESULTS: SIEs of 86% +/- 3% and 97% +/- 3% in nonischemic, and 25% +/- 5% and 29% +/- 8% in ischemic regions were found within three minutes of onset of ischemia with Gd(ABE-DTTA) and Gd(DTPA), respectively. For the rest of the 30 minutes of ischemia, with Gd(ABE-DTTA) SIE of 60% +/- 3% and 25% +/- 5% persisted in the nonischemic and ischemic regions, respectively. With Gd(DTPA), however, SIE in the nonischemic areas decreased rapidly after the first three minutes of ischemia, while SIE in the ischemic areas increased, abolishing contrast. Thus, there was a persistent contrast with Gd(ABE-DTTA) and a short-lived contrast with Gd(DTPA) during ischemia. Furthermore, with Gd(ABE DTTA) some contrast was still visible in the early reperfusion period. CONCLUSION: Gd(ABE-DTTA) in an ischemia/reperfusion model induces a persistent MRI contrast between regions of normal and ischemic myocardium, and verifies reperfusion. Therefore, it can be used to detect myocardial ischemic events. PMID- 15834917 TI - Predicting final infarct size using acute and subacute multiparametric MRI measurements in patients with ischemic stroke. AB - PURPOSE: To identify early MRI characteristics of ischemic stroke that predict final infarct size three months poststroke. MATERIALS AND METHODS: Multiparametric MRI (multispin echo T2-weighted [T2W] imaging, T1-weighted [T1W] imaging, and diffusion-weighted imaging [DWI]) was performed acutely (<24 hours), subacutely (three to five days), and at three months. MRI was processed using maps of apparent diffusion coefficient (ADC), T2, and a self-organizing data analysis (ISODATA) technique. Analyses began with testing for individual MRI parameter effects, followed by multivariable modeling with assessment of predictive ability (R(2)) on final infarct size. RESULTS: A total of 45 patients were studied, 15 of whom were treated with tissue plasminogen activator (tPA) before acute MRI. The acute DWI and DWI-ISODATA mismatch lesion size, and the interactions of ADC, T2, and T2W imaging lesion with tPA remained in the final multivariable model (R(2) = 70%). A large acute DWI lesion or DWI < ISODATA lesion independently predicted increase in the final infract size, with predictive ability 68%. Predictive ability increased (R(2) = 83%) when subacute MRI parameters were included along with acute DWI, DWI-ISODATA mismatch, and acute T2W image lesion size by tPA treatment interaction. Subacute DWI > acute DWI lesion size predicted an increased final infarct size (P < 0.01). CONCLUSION: Acute-phase DWI and DWI-ISODATA mismatch strongly predict the final infarct size. An acute-to-subacute DWI lesion size change further increases the predictive ability of the model. PMID- 15834918 TI - Normal white matter development from infancy to adulthood: comparing diffusion tensor and high b value diffusion weighted MR images. AB - PURPOSE: To evaluate the sensitivity of high b value diffusion weight magnetic resonance imaging (DWI) in detecting normal white matter maturation, compare it to conventional diffusion tensor imaging (DTI), and to obtain normative quantitative data using this method. MATERIALS AND METHODS: High b value DWI (b(max) = 6000 sec/mm(2)) using q-space analysis and conventional DTI (b = 1000 sec/mm(2)) were performed on 36 healthy subjects aged 4 months to 23 years. Fractional-anisotropy (FA), apparent-displacement, and apparent-probability values were measured in all slices and in six regions of interest (ROIs) of large fiber tracks. Values were correlated with each other and with age using regression analysis. RESULTS: FA, displacement, and probability indices from all slices were highly correlated with each other (r > 0.87, P < 0.0001) and with age (r > 0.82, P < 0.0001). All age-related changes in the six pre-determined ROIs were best fitted by mono-exponential functions. Changes in the splenium extended to a later age when compared with the genu of the corpus-callosum, while the centrum semi-ovale demonstrated the latest changes with age. CONCLUSIONS: High b value DWI and DTI showed changes in white matter from infancy through adulthood. However, high b-value detects a signal that is likely to originate mainly from the intra-axonal water population, and thus may represent different aspects of development and different sensitivity to pathology. PMID- 15834919 TI - Rapid measurement of renal artery blood flow with ungated spiral phase-contrast MRI. AB - PURPOSE: To verify the potential of ungated spiral phase-contrast (USPC), which has been shown to provide accurate and reproducible time-averaged measurements of pulsatile flow, for rapid measurement of renal artery blood flow (RABF) in vivo. MATERIALS AND METHODS: The RABF rates of 11 normal human subjects and one patient with renal failure were measured with USPC within six seconds. RESULTS: Rapid USPC scans produced reproducible RABF measurements (SD < or = 9%) that agreed with the normal RABF rates known from the literature. The RABF rates of the patient with renal failure were substantially less (<50-65%) than the normal RABF rates. CONCLUSION: The results demonstrate that it is now possible to obtain rapid and consistent RABF measurements within six seconds with USPC. PMID- 15834920 TI - Whole-body MR angiography using a novel 32-receiving-channel MR system with surface coil technology: first clinical experience. AB - PURPOSE: To demonstrate the feasibility of detecting atherosclerotic vascular disease using an innovative magnetic resonance angiography (MRA) protocol in combination with a dedicated whole-body MR scanner with new surface coil technology. MATERIALS AND METHODS: A total of 10 volunteers and eight patients with peripheral arterial occlusive disease (PAOD) were examined at 1.5 T. Conventional digital subtraction angiography (DSA) of the symptomatic region was available as a reference standard in all eight patients. Depending on subjects' size, four to five three-dimensional data sets were acquired using an adapted injection protocol. Images were assessed independently by two readers for vascular pathology. Additionally, signal-to-noise ratios (SNRs) and contrast-to noise ratios (CNRs) were measured. RESULTS: Whole-body MRA yielded excellent sensitivity and specificity of more than 95% for both readers with high interobserver agreement (k = 0.93). Surface coil signal reception rendered a high SNR (mean 151.28 +/- 54.04) and CNR (mean 120.75 +/- 46.47). Despite lower SNR and CNR of the cranial and cervical vessels, a two-step injection protocol exhibited less venous superposition and therefore proved to be superior compared to single-bolus injection. CONCLUSION: Our approach provides accurate noninvasive high-resolution imaging of systemic atherosclerotic disease, covering the arterial vasculature from intracranial arteries to distal runoff vessels. The recently introduced MR scanner and coil technology is feasible to significantly increase the performance of whole-body MRA. PMID- 15834921 TI - Functional magnetic resonance imaging of the human lumbar spinal cord. AB - PURPOSE: To determine whether consistent regions of activity could be observed in the lumbar spinal cord of single subjects with spin-echo functional MRI (fMRI) if several repeated experiments were performed within a single imaging session. MATERIALS AND METHODS: Repeated fMRI experiments of the human lumbar spinal cord were performed at 1.5 T with a single-shot spin-echo technique (half-Fourier single-shot turbo spin-echo (HASTE)) as used by previous investigators, and a modified method (fluid-attenuated inversion recovery (FLAIR)-HASTE) that nulled the otherwise highly variable signal from the cerebrospinal fluid (CSF). RESULTS: FLAIR-HASTE reduced the variability of the signal in the CSF region to background levels, and presumably reduced associated artifacts in the spinal cord. Consistent areas of activation in the spinal cord in response to a thermal stimulus just below the knee were not observed across the fMRI experiments with either method. CONCLUSION: FLAIR-HASTE was useful for removing artifact in the spinal cord signal induced by variability in the CSF signal. However, with the techniques used in this study, we were not able to confirm the presence of a consistent fMRI response in the lumbar spinal cord because of the signal enhancement by extravascular protons (SEEP) effect during thermal stimulation of the hindlimb. PMID- 15834922 TI - Comparison of qualitative and quantitative measurements on unenhanced T1-weighted fat saturation MR images in predicting pancreatic pathology. AB - PURPOSE: To evaluate the accuracy of signal intensity (SI) analysis on unenhanced fat-suppressed T1-weighted MR images in the diagnosis of pancreatic disease and to compare subjective interpretation with different quantitative measurements. MATERIALS AND METHODS: The pancreas was evaluated in 159 patients (86 normal and 73 with pancreatic disease) with spoiled gradient echo (GRE) T1-weighted fat saturation MR images. The relative SI of the pancreas to liver and spleen was quantitatively measured using regions of interest (ROIs) and qualitatively assessed by two independent observers. RESULTS: The mean values between a normal and an abnormal pancreas with pancreas-liver ratios of 0.14 +/- 0.37 vs. -0.32 +/ 0.24, respectively, and pancreas-spleen ratios of 0.89 +/- 0.55 vs. 0.02 +/- 0.43, respectively, were significantly different (P < 0.001). The pancreas-liver SI ratio was significantly better than the pancreas-spleen ratio throughout the disease group (area under the receiver operating characteristic (ROC) curve +/- SD; 0.92 +/- 0.02 for pancreas-liver vs. 0.86 +/- 0.03 for pancreas-spleen, P < 0.01), and after excluding cases of acute pancreatitis (0.96 +/- 0.02 for pancreas-liver vs. 0.89 +/- 0.03 for pancreas-spleen, P < 0.01). There was no statistically significant difference between quantitative and qualitative analysis (area under the ROC curve +/- SD; 0.93 +/- 0.02 vs. 0.93 +/- 0.02 for the entire disease group; excluding acute pancreatitis 0.96 +/- 0.02 vs 0.97 +/- 0.02) for the diagnosis of pancreatic disease when using liver as internal standard. The interobserver concordance was very good (kappa > 0.71). The sensitivity of visual liver comparison was 80% in the entire disease group and 91% after the cases of acute pancreatitis were excluded, while specificity was 93%. CONCLUSION: The pancreas-liver ratio is the best quantitative means of distinguishing normal from abnormal pancreas. Visual observation by experienced observers (qualitative measurement) was just as accurate as quantitative measurement. Detection of pancreatic pathology can be made with high accuracy by visually comparing the SI of the pancreas with that of the normal liver. PMID- 15834923 TI - 3D-Epitope-Explorer (3DEX): localization of conformational epitopes within three dimensional structures of proteins. AB - Neutralizing antibodies often recognize conformational, discontinuous epitopes. Linear peptides mimicking such conformational epitopes can be selected from phage display peptide libraries by screening with the respective antibodies. However, it is difficult to localize these "mimotopes" within the three-dimensional (3D) structures of the target proteins. Knowledge of conformational epitopes of neutralizing antibodies would help to design antigens able to elicit protective immune responses. Therefore, we provide here a software that allows to localize linear peptide sequences within 3D structures of proteins. The 3D-Epitope Explorer (3DEX) software allows to map conformational epitopes in 3D protein structures based on an algorithm that takes into account the physicochemical neighborhood of C(alpha)- or C(beta)-atoms of individual amino acids. A given amino acid of a peptide sequence is localized within the protein and the software searches within predefined distances for the amino acids neighboring that amino acid in the peptide. Surface exposure of the amino acids can also be taken into consideration. The procedure is then repeated for the remaining amino acids of the peptide. The introduction of a joker function allows to map peptide mimotopes, which do not necessarily have 100% sequence homology to the protein. Using this software we were able to localize mimotopes selected from phage displayed peptide libraries with polyclonal antibodies from HIV-positive patient plasma within the 3D structure of gp120, the exterior glycoprotein of HIV-1. We also analyzed two recently published peptide sequences corresponding to known conformational epitopes to further confirm the integrity of 3DEX. PMID- 15834924 TI - Quantum chemical modeling of CO oxidation by the active site of molybdenum CO dehydrogenase. AB - The catalytic mechanism of molybdenum containing CO dehydrogenase has been studied using hybrid DFT methods with quite large chemical models. The recent high-resolution X-ray structure, showing the surprising presence of copper linked to molybdenum, was used as a starting point. A pathway was initially found with a low barrier for C-O bond formation and CO2 release. However, this pathway did not include the formation of any S-CO2 species, which had been suggested by experiments with an n-butylisocyanide inhibitor. When these SCO2 structures were studied they were found to lead to deep minima, making CO2 release much more difficult. A large effort was spent, including investigations of other spin states, varying the number of protons and electrons, adding water, etc., until a plausible pathway for S-C bond cleavage was found. In this pathway a water molecule is inserted in between molybdenum and the SCO2 group. Full catalytic cycles, including electron and proton transfers, are constructed both with and without S-C bond formation. When these pathways are extended to two full catalytic cycles it can be understood why the formation of the S-C bond actually makes catalysis faster, even though the individual step of CO2 release becomes much more difficult. These results agree well with experimental findings. PMID- 15834925 TI - Polymorphisms in GLTSCR1 and ERCC2 are associated with the development of oligodendrogliomas. AB - BACKGROUND: Deletions of 19q have been associated with gliomas, especially oligodendrogliomas. In addition, cases with oligodendrogliomas with the 19q deletion have been observed to have a better survival compared with cases without the 19q deletion. The authors have previously described a 150-kilobase minimal deletion region in gliomas that maps to 19q13.33 and contains 3 novel candidate genes (GLTSCR1, EHD2, and GLTSCR2). METHODS: The authors performed an association study using 141 cases with gliomas (61 cases with astrocytomas, 40 cases with oligodendrogliomas, 40 cases with mixed oligoastrocytomas) and 108 general controls. They evaluated 7 single nucleotide polymorphisms (SNPs) in 6 genes within and nearby the minimal 19q deletion region (ERCC2, RAI, ASE-1, ERCC1, GLTSCR1, and LIG1). RESULTS: The prevalence of a germline GLTSCR1-exon-1 T allele (SNP rs1035938) was 40% in cases with oligodendrogliomas compared with 27% in controls (P = 0.029), and the prevalence of an ERCC2-exon-22 T allele (SNP rs1052555) was 35% in cases with oligodendrogliomas compared with 18% in controls (P = 0.043). One high-risk and 1 low-risk haplotype were associated with oligodendroglioma development (P = 0.003 and 0.026, respectively). Cases with oligodendrogliomas with the 19q deletion had a significantly higher frequency of the GLTSCR1-exon-1 T allele compared with cases without the 19q deletion (P = 0.01). It was noteworthy that cases with gliomas who were homozygous for the GLTSCR1-exon-1 T allele had a significantly better survival: 77% and 68% survival at 2 and 5 years compared with 56% and 34% for other genotypes (P = 0.02, log rank test). Multivariable analysis identified grade, age, and the GLTSCR1-exon-1 and ERCC2-exon-22 genotypes as independent predictors for survival. CONCLUSIONS: These results suggested that alterations in GLTSCR1 (or a closely linked gene) were associated with the development and progression of oligodendroglioma. PMID- 15834926 TI - Acute and late morbidity in the treatment of advanced bladder carcinoma with accelerated radiotherapy, carbogen, and nicotinamide. AB - BACKGROUND: Accelerated radiotherapy combined with carbogen and nicotinamide (ARCON) to overcome tumor hypoxia and cell proliferation achieved high tumor control and survival in Phase II studies of patients with advanced head and neck and bladder carcinomas. Thus, morbidity and treatment outcomes from the latter study were analyzed to evaluate the therapeutic potential of ARCON. METHODS: Acute and late morbidity was assessed in 105 patients with high-grade superficial or muscle-invasive bladder carcinoma who were given accelerated radiotherapy (50 55 grays in 4 weeks) with carbogen alone or with ARCON. Urinary dysfunction was scored based on daytime frequency, nocturia, incontinence, dysuria, hematuria, and urgency. Bowel morbidity was based on stool frequency and consistency, rectal discharge, blood loss, and medication. Endpoints for treatment outcome were overall survival, disease-free survival, and locoregional control. RESULTS: Nearly all patients experienced reduced ability to retain urine beyond 2 hours, although 20-30% had almost normal function at night. Incidence of acute moderate or worse dysuria was 41% with ARCON and 56% with carbogen; 96% and 76% of patients, respectively, had bowel frequencies > or = 3 times per day. By 10-12 weeks from the start of radiotherapy, acute reactions returned to baseline levels. At 3 years, the daytime frequency < or = 2 times per hour was approximately 75% in both arms. Incidence of severe hematuria (< or = 25%) and urinary urgency (< or = 16%) was much lower. No more than 6% of patients had severe bowel morbidity. With most assays, the differences between schedules were not significant either for acute or late morbidity. Local tumor control and survival rates at 3 years were 53% and 43%, respectively, for ARCON, similar to the rates for carbogen alone. CONCLUSIONS: Historical comparisons suggested no overt increase in normal tissue radiosensitivity when adding carbogen and nicotinamide. Although, for some endpoints, the incidence of late sequelae was higher than expected, overall morbidity was no worse than reported by others. The data indicated that ARCON could achieve a therapeutic gain in patients with advanced bladder carcinoma. A Phase III, randomized, multicenter trial is underway currently in the United Kingdom to evaluate these findings. PMID- 15834927 TI - Hyperhomocysteinemia and the MTHFR C677T polymorphism promote steatosis and fibrosis in chronic hepatitis C patients. AB - The factors and mechanisms implicated in the development of hepatitis C virus (HCV)-related steatosis are unknown. Hyperhomocysteinemia causes steatosis, and the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism induces hyperhomocysteinemia. We investigated the role of these factors in the development of HCV-related steatosis and in the progression of chronic hepatitis C (CHC). One hundred sixteen CHC patients were evaluated for HAI, fibrosis and steatosis grades, body mass index, HCV genotypes, HCV RNA levels, homocysteinemia, and the MTHFR C677T polymorphism. Hyperhomocysteinemia was associated with the TT genotype of MTHFR (r = 0.367; P = .001). Median values of homocysteine in the CC, CT, and TT genotypes of the MTHFR gene were 9.3, 12.2, and 18.6 micromol/L, respectively (P = .006). Steatosis correlated with the MTHFR polymorphism, homocysteinemia, HAI and fibrosis. Steatosis above 20% was significantly associated with fibrosis. Prevalence and high grade (>20%) of steatosis were 41% and 11% in CC, 61% and 49% in CT, and 79% and 64% in TT, respectively (P = .01). Relative risk of developing high levels of steatosis was 20 times higher for TT genotypes than CC genotypes. According to multivariate analysis, steatosis was independently associated with hyperhomocysteinemia (OR = 7.1), HAI (OR = 3.8), liver fibrosis (OR = 4.0), and HCV genotype 3 (OR = 4.6). On univariate analysis, fibrosis was associated with age, steatosis, MTHFR, homocysteinemia and HAI; however, on multivariate analysis, liver fibrosis was independently associated with age (P = .03), HAI (P = .0001), and steatosis (P = .007). In conclusion, a genetic background such as the MTHFR C677T polymorphism responsible for hyperhomocysteinemia plays a role in the development of higher degree of steatosis, which in turn accelerates the progression of liver fibrosis in CHC. PMID- 15834928 TI - HER-2/neu expression as a predictor of response to neoadjuvant docetaxel in patients with operable breast carcinoma. AB - BACKGROUND: The use of biologic markers to predict response to neoadjuvant chemotherapy may permit tailoring regimens to achieve maximal tumor response. Taxanes have demonstrated excellent activity in breast carcinoma; however, tumor specific factors that predict clinical response have not been characterized thoroughly. METHODS: The authors performed a historic review evaluating the association of tumor prognostic factors and response to neoadjuvant cyclophosphamide and doxorubicin (AC) with or without docetaxel (D) (AC vs. AC+D) in 121 women who previously were enrolled in a Phase III, randomized, clinical trial. Using pretreatment biopsy materials, immunohistochemical studies were performed for estrogen receptor (ER), progesterone receptor (PR), HER-2/neu, p53, and Ki-67. Outcome variables were pathologic complete response (pCR) and positive clinical response (cPOS), which was defined as a >/= 50% regression in clinical tumor size prior to surgery. RESULTS: In a multivariate analysis that controlled for tumor size and lymph node status, improved cPOS rates were observed with the addition of docetaxel in women with HER-2/neu-negative tumors (81% vs. 51%; P < 0.05), yielding an adjusted odds ratio of 3.5 (95% confidence interval, 1.2-13.0) in favor of docetaxel. Women who had HER-2/neu-negative tumors appeared to have a lower response rate with AC alone compared with women who had HER-2/neu-positive tumors (51% vs. 75%; P = 0.06), but response rates were matched when docetaxel was added (81% vs. 78%; P = 0.99). ER, PR, p53, and Ki-67 results were not associated significantly with response rates. CONCLUSIONS: HER-2/neu status may predict improved clinical response rates from the addition of docetaxel to anthracycline-based neoadjuvant chemotherapy. Docetaxel may "rescue" the response in women who have HER-2/neu-negative tumors to match that observed in women who have HER-2/neu-positive tumors treated with AC alone. PMID- 15834929 TI - Secretin activation of the apical Na+-dependent bile acid transporter is associated with cholehepatic shunting in rats. AB - The role of the cholangiocyte apical Na(+)-dependent bile acid transporter (ASBT) in bile formation is unknown. Bile acid absorption by bile ducts results in cholehepatic shunting, a pathway that amplifies the canalicular osmotic effects of bile acids. We tested in isolated cholangiocytes if secretin enhances ASBT translocation to the apical membrane from latent preexisting intracellular stores. In vivo, in bile duct-ligated rats, we tested if increased ASBT activity (induced by secretin pretreatment) results in cholehepatic shunting of bile acids. We determined the increment in taurocholate-dependent bile flow and biliary lipid secretion and taurocholate (TC) biliary transit time during high ASBT activity. Secretin stimulated colchicine-sensitive ASBT translocation to the cholangiocyte plasma membrane and (3)H-TC uptake in purified cholangiocytes. Consistent with increased ASBT promoting cholehepatic shunting, with secretin pretreatment, we found TC induced greater-than-expected biliary lipid secretion and bile flow and there was a prolongation of the TC biliary transit time. Colchicine ablated secretin pretreatment-dependent bile acid-induced choleresis, increased biliary lipid secretion, and the prolongation of the TC biliary transit. In conclusion, secretin stimulates cholehepatic shunting of conjugated bile acids and is associated with increased cholangiocyte apical membrane ASBT. Bile acid transport by cholangiocyte ASBT can contribute to hepatobiliary secretion in vivo. PMID- 15834930 TI - Direct correlation between adsorption-induced changes in protein structure and platelet adhesion. AB - It is widely recognized that adsorbed proteins on biomaterial surfaces tend to initiate thrombus formation, although the specific mechanisms involved are still not well understood. In attempts to decrease the conformational change of adsorbed proteins, surface treatments that reduce surface hydrophobicity have been considered, such as the sulfonation of low-density polyethylene and isotactic polypropylene. The objectives of this present research were to study how changes in surface chemistry influence the degree of conformational change of adsorbing proteins and to investigate the correlation between the change in adsorbed protein structure and platelet response. Adsorbed porcine serum albumin and porcine fibrinogen were used as the model proteins for determining the effects of sulfonation on protein conformational change. Circular dichroism spectroscopy studies showed that the proteins were less altered structurally on the sulfonated surfaces. Platelet adhesion studies were used to correlate the number of adhered platelets with the amount of conformational change in adsorbed proteins on the polymer surface. The results of these studies show a linear correlation between platelet adhesion and the degree of adsorption-induced protein conformational change. These findings suggest that the degree of protein conformational change after adsorption is a dominant mechanism governing platelet interactions with biomaterial surfaces. PMID- 15834931 TI - Effects of bone morphogenetic protein-2 and hyaluronic acid on the osseointegration of hydroxyapatite-coated implants: an experimental study in sheep. AB - Research efforts aim at enhancing early osseointegration of cementless implants to improve early fixation and, thus, reduce the risk of loosening. The aim of the present study was to investigate whether bone morphogenetic protein (BMP) 2 had a positive effect on the osseointegration of hydroxyapatite-coated implants. Hydroxyapatite (HA) implants (perforated hollow cylinders and solid rods) were coated with BMP-2 and hyaluronic acid (HY) as the carrier or with HY alone. Uncoated HA implants served as controls. The osseointegration of the implants was evaluated either by light microscopy or by pullout tests after 1, 2, and 4 weeks of unloaded implantation in the cancellous bone of 24 sheep. The BMP-2 coating significantly increased bone growth into the implant perforations compared with HA-coated implants at 2 and 4 weeks. Bone-implant contact and interface shear strength of BMP-2 implants were lower than HA implants at 2 weeks. At 4 weeks, there was no significant difference in bone-implant contact and shear strength between BMP-2 and HA-coated implants. The BMP-2 coating enhanced gap healing but had no positive or even an inhibitory effect (at 2 weeks) on bone-implant contact and interface shear strength. In the clinical situation, a perfect press-fit implantation cannot be achieved, and BMP-2 may be beneficial for enhancing bone growth into gaps around cementless implants. PMID- 15834932 TI - The viability and function of primary rat hepatocytes cultured on polymeric membranes developed for hybrid artificial liver devices. AB - Bioartificial liver devices require membranes to support the function and viability of hepatocytes because they are anchorage-dependent cells. This study investigated the ability of several polymeric membranes to support the functions of primary hepatocyte cultures. Tailor-made membranes were sought by synthesizing acrylonitrile copolymers with different comonomers resulting in ionic, hydrophilic, or reactive functional groups on the polymer surface. Hepatocyte morphology and viability were assessed by confocal microscopy, and function by the content and activities of cytochrome P450, and the expression of glutathione S-transferases. Hydrophilic membranes (polyacrylonitrile and acrylonitrile copolymerized with 2-acrylamino-2-methyl-propane sulfonic acid) were more biocompatible than hydrophobic membranes such as polysulfone. The chemistry of the hydrophilic group was important; amine groups had a deleterious effect on maintenance of the primary hepatocytes. The biocompatibility of hydrophobic membranes was improved by collagen coating. Improving the chemistry of membranes for artificial liver devices will enhance the phenotypic stability of the cells, enabling us to prolong treatment times for patients. PMID- 15834933 TI - Rheological and recovery properties of poly(ethylene glycol) diacrylate hydrogels and human adipose tissue. AB - The viscosity and elastic and viscous moduli of poly(ethylene glycol) diacrylate (PEGDA) hydrogels and human abdominal adipose tissue are measured as a function of shear rate and frequency. Results indicate that both materials exhibit shear thinning and are viscoelastic in nature. Rheological tests suggest that the hydrogels become firmer as strain and frequency increase. Adipose tissue, however, begins to fail at higher strains and frequencies. This behavior is confirmed by measuring the complex modulus of both materials as a function of strain. Recovery properties are also measured for each material as a function of deformation. Although PEGDA hydrogels are able to recover up to 78% of their original height after 15% deformation, adipose tissue is not able to recover over the range of deformations tested. The frequencies and strains over which the tests are conducted are those physiologically experienced by the human body. The hydrogels are able to withstand this range of forces and, hence, are appropriate for use as a soft tissue filler material. In addition, the hydrogels swell 38.1% +/- 0.9% independent of surface area. The complex modulus of hydrogels of varying polymer concentrations is also measured as a function of strain to determine the effects of changing polymer content. These results indicate that as polymer content increases, the hydrogels become firmer due to the higher number of polymer chains and behave more elastically. PMID- 15834934 TI - Adjuvant iodine-131-labeled lipiodol for prevention of intrahepatic recurrence of hepatocellular carcinoma: which is the best treatment schedule? PMID- 15834935 TI - Changes in gallbladder bile composition and crystal detection time in morbidly obese subjects after bariatric surgery. AB - The aim of the present study was to elucidate the mechanisms of development of cholesterol crystals and gallstones during weight reduction in obese subjects. Twenty-five morbidly obese, gallstone-free subjects underwent vertical-banded gastroplasty. Gallbladder bile was collected at the time of the operation via needle aspiration and 1.1-7.3 months after the operation via ultrasound-guided transhepatic puncture of the gallbladder. The mean weight loss was 17 kg. Two patients developed gallstones and 10 patients displayed cholesterol crystals in their bile. In patients with a follow-up time of less than 2 months (n = 13), cholesterol saturation increased from 90% to 114% but tended to decrease in the patients with a follow-up time of more than 2 months. The extraction of the concanavalin-A-binding fraction from gallbladder bile obtained after weight reduction in 7 patients prolonged crystallization detection time from 6 to 10 days. The hexosamine concentration, a marker for mucin, was increased by about 100% in bile obtained in 6 of 7 patients after weight reduction. In conclusion, the results indicate that crystallization-promoting compounds (mucin) are of great importance in the development of cholesterol crystals and gallstones in obese subjects during weight reduction, probably because of defective gallbladder emptying. PMID- 15834936 TI - Transcriptome profiling of individual larvae of two different developmental modes in the poecilogonous polychaete Streblospio benedicti (Spionidae). AB - Understanding the range of biochemical and physiological phenotypes in a cohort of embryos or larvae is crucial to understanding the lifespan, dispersal potential, and recruitment success of the early life history stages of a species. In this study, a novel kinetic assay has been employed to profile the transcriptome pool complexity in individual larvae of both planktotrophic and lecithotrophic developmental modes in the poecilogonous polycheate Streblospio benedicti. Using a nano-scale synthesis strategy, the mRNA pool in a single embryo or larva can be amplified into cDNA for quantitative characterization in a high-throughput, kinetic reannealing assay in a 96-well, microtiterplate format. This assay generates transcript-pool complexity estimates at 1 degrees C temperature increments for each sample producing 3,360 quantitative measurements per 96-well plate. Measuring transcriptome complexity on 8 individual planktotrophic and 8 individual lecithotrophic larvae (with 4 duplicate assays for each individual) reveals a more complex gene expression profile in planktotrophic larvae and a lower level of interindividual variation in expression patterns in lecithotrophic larvae. Although differences in these gene expression patterns are more likely due to physiological differences between feeding and non-feeding larval types in these late-stage individuals, this is one of the first assessments of inter-individual variation in gene expression patterns in marine invertebrate larvae and indicates a large potential for developmental variability. PMID- 15834937 TI - MELD score and clinical type predict prognosis in hepatorenal syndrome: relevance to liver transplantation. AB - Important progress has been made recently regarding the pathogenesis and treatment of hepatorenal syndrome (HRS). However, scant information exists about factors predicting outcome in patients with cirrhosis and HRS. Moreover, the prognostic value of the model of end-stage liver disease (MELD) score has not been validated in the setting of HRS. The current study was designed to assess the prognostic factors and outcome of patients with cirrhosis and HRS. The study included 105 consecutive patients with HRS. Forty-one patients had type 1 HRS, while 64 patients had type 2 HRS. Patients with type 1 HRS not only had more severe liver and renal failure than type 2 patients, they also had greater impairment of circulatory function, as indicated by lower arterial pressure and higher activation of vasoconstrictor factors. In the whole series, the median survival was 3.3 months. In a multivariate analysis of survival, only HRS type and MELD score were associated with an independent prognostic value. All patients with type 1 HRS had a high MELD score (> or =20) and showed an extremely poor outcome (median survival: 1 mo). By contrast, the survival of patients with type 2 HRS was longer and dependent on MELD score (> or =20, median survival 3 mo; <20, median survival 11 mo; P < .002). In conclusion, the outcome of patients with cirrhosis and HRS can be estimated by using two easily available variables, HRS type and MELD score. These data can be useful in the management of patients with HRS, particularly for patients who are candidates for liver transplantation. PMID- 15834938 TI - Non-neural ectoderm is really neural: evolution of developmental patterning mechanisms in the non-neural ectoderm of chordates and the problem of sensory cell homologies. AB - In chordates, the ectoderm is divided into the neuroectoderm and the so-called non-neural ectoderm. In spite of its name, however, the non-neural ectoderm contains numerous sensory cells. Therefore, the term "non-neural" ectoderm should be replaced by "general ectoderm." At least in amphioxus and tunicates and possibly in vertebrates as well, both the neuroectoderm and the general ectoderm are patterned anterior/posteriorly by mechanisms involving retinoic acid and Hox genes. In amphioxus and tunicates the ectodermal sensory cells, which have a wide range of ciliary and microvillar configurations, are mostly primary neurons sending axons to the CNS, although a minority lack axons. In contrast, vertebrate mechanosensory cells, called hair cells, are all secondary neurons that lack axons and have a characteristic eccentric cilium adjacent to a group of microvilli of graded lengths. It has been highly controversial whether the ectodermal sensory cells in the oral siphons of adult tunicates are homologous to vertebrate hair cells. In some species of tunicates, these cells appear to be secondary neurons, and microvillar and ciliary configurations of some of these cells approach those of vertebrate hair cells. However, none of the tunicate cells has all the characteristics of a hair cell, and there is a high degree of variation among ectodermal sensory cells within and between different species. Thus, similarities between the ectodermal sensory cells of any one species of tunicate and craniate hair cells may well represent convergent evolution rather than homology. PMID- 15834939 TI - Expression of engrailed in the developing brain and appendages of the onychophoran euperipatoides kanangrensis (Reid). AB - We have cloned an engrailed-class gene in the onychophoran Euperipatoides kanangrensis and investigated its expression using in situ hybridisation. The expression pattern was found to differ drastically from that previously described for another onychophoran species. In the present investigation, engrailed transcripts were detected in a subset of developing neurons in the brain anlage, and in the mesoderm as well as ectoderm of the developing limb buds. The engrailed positive cells of the brain are of differing developmental maturity, ranging from subepidermal neuronal precursors to neurons located basally in the embryo with developing axons. The lack of the traditional expression in the posterior compartment of segments reported earlier in onychophorans is discussed, and we suggest that onychophorans may have acquired two copies of engrailed with different functions. PMID- 15834940 TI - AGR2, an androgen-inducible secretory protein overexpressed in prostate cancer. AB - AGR2, the human homologue of Xenopus anterior gradient 2 (XAG2), was identified by a suppression subtractive hybridization-based technique as an androgen inducible gene. There are two AGR2 transcripts, which encode the same secretory protein of 175 amino acids. The androgen induction was time- and dose-dependent, with more than a 10-fold increase in the level of AGR2 mRNA after 48 hr of treatment with 10(-9) M R1881. Expression of AGR2 mRNA was specifically detected in limited human tissue rich in epithelial cells, including the prostate gland. Analysis of 46 microdissected primary prostate adenocarcinoma samples showed that AGR2 mRNA expression was markedly elevated in the majority of tumors as compared to matched adjacent benign tissues. Androgen-induced AGR2 protein expression was demonstrated in LNCaP cells by Western blot analysis with an anti-AGR2 antibody. Immunohistochemistry analysis indicated that AGR2 protein expression was highly restricted to the secretory epithelial cells in the prostate gland. In tissue sections from radical prostatectomy specimens, immunohistochemical staining of AGR2 showed markedly increased expression in high-grade prostatic intraepithelial neoplasia and Gleason pattern 3-4 prostatic adenocarcinoma. Therefore, the androgen-induced secretory protein AGR2 may serve as a potential therapeutic target and/or molecular marker for prostate cancer. PMID- 15834941 TI - Identification of androgen-responsive genes that are alternatively regulated in androgen-dependent and androgen-independent rat prostate tumors. AB - The vast majority of androgen-dependent prostate tumors progress toward incurable, androgen-independent tumors. The identification of androgen-responsive genes, which are still actively transcribed in the tumors of patients who have undergone androgen ablation, may shed light on the molecular mechanisms underlying this phenomenon. To address this question, we chose the Dunning R3327 rat model system, in which the progression from androgen-dependent to independent tumors is represented by several transplantable prostate-derived tumors. Gene expression profiles were analyzed in normal rat prostates and in the prostates of rats 14 days after castration by use of microarrays containing approximately 5,000 oligonucleotides, together representing more than 4,800 known rat genes. These expression profiles were compared with similarly obtained expression profiles of androgen-dependent and androgen-independent rat prostate tumors. By doing so, a series of known and novel prostate cancer-associated androgen-responsive genes was identified. Within this series, we were able to identify several clusters of genes that are differentially regulated in the various prostate tumors. These genes may serve as (i) novel prognostic identifiers and (ii) novel therapeutic targets. PMID- 15834942 TI - Cytogenetic alterations in nonmelanoma skin cancer: a review. AB - Since the advent of cytogenetic analysis, knowledge about fundamental aspects of cancer biology has increased, allowing the processes of cancer development and progression to be more fully understood and appreciated. Classical cytogenetic analysis of solid tumors had been considered difficult, but new advances in culturing techniques and the addition of new cytogenetic technologies have enabled a more comprehensive analysis of chromosomal aberrations associated with solid tumors. Our purpose in this review is to discuss the cytogenetic findings on a number of nonmelanoma skin cancers, including squamous- and basal cell carcinomas, keratoacanthoma, squamous cell carcinoma in situ (Bowen's disease), and solar keratosis. Through classical cytogenetic techniques, as well as fluorescence-based techniques such as fluorescence in situ hybridization and comparative genomic hybridization, numerous chromosomal alterations have been identified. These aberrations may aid in further defining the stages and classifications of nonmelanoma skin cancer and also may implicate chromosomal regions involved in progression and metastatic potential. This information, along with the development of newer technologies (including laser capture microdissection and comparative genomic hybridization arrays) that allow for more refined analysis, will continue to increase our knowledge about the role of chromosomal events at all stages of cancer development and progression and, more specifically, about how they are associated with nonmelanoma skin cancer. PMID- 15834943 TI - Functional evidence of decreased tumorigenicity associated with monochromosome transfer of chromosome 14 in esophageal cancer and the mapping of tumor suppressive regions to 14q32. AB - Despite the abundant evidence of high allelic loss of chromosome arm 14q in human cancers, tumor-suppressor genes mapped to this chromosome have yet to be identified. To narrow the search for candidate genes, we performed monochromosome transfer of chromosome 14 into an esophageal carcinoma cell line, SLMT-1 S1. Statistically significant suppression of the tumorigenic potential of microcell hybrids containing the transferred chromosome 14 provided functional evidence that tumor-suppressive regions of chromosome 14 are essential for esophageal cancer. Tumor segregants emerging in nude mice during the tumorigenicity assay were analyzed by detailed PCR-microsatellite typing to identify critical nonrandomly eliminated regions (CRs). A 680-kb CR mapped to 14q32.13 and an approximately 2.2-Mb CR mapped to 14q32.33 were delineated. Dual-color BAC FISH analysis of microcell hybrids and tumor segregants verified the selective loss of the 14q32.13 region. In contrast, similar transfers of an intact chromosome 11 into SLMT-1 S1 did not significantly suppress tumor formation. These functional complementation studies showing the correlation of tumorigenic potential with critical regions of chromosome 14 validated the importance of the 14q32 region in tumor suppression in esophageal cancer. The present study also paved the path for further identification of novel tumor-suppressor genes that are relevant to the molecular pathogenesis of esophageal cancer. PMID- 15834944 TI - Detection of chromosomal imbalances in retinoblastoma by matrix-based comparative genomic hybridization. AB - The genetic hallmark of retinoblastoma is mutation or deletion of the RB1 gene, whereas other genetic alterations that are also required are largely unknown. To screen for genomic imbalances on a genomewide level, we studied a series of 17 primary retinoblastomas by matrix-based comparative genomic hybridization (matrix CGH). The matrix-CGH chip contained 6,000 immobilized genomic DNA fragments covering the human genome, with an average resolution of about 500 kb. The most frequent imbalances detected were gains on chromosome arms 1q (12 of 17), 6p (10 of 17), 2p (5 of 17), and 19q (4 of 17) and loss on 16q (7 of 17). Candidate regions could be narrowed to small intervals by the identified minimally overlapping regions on 1q22, 1q32.1q32.2, 2p24.1, and 6p21.33-p21.31. Furthermore, two as-yet-unknown high-level amplifications were detected, each in a single patient, on chromosome bands 1p34.2 and 1p33. Thus, this study identified new chromosomal regions and therefore potential candidate genes that may play a role in retinoblastoma. PMID- 15834945 TI - Intratumor versus intertumor heterogeneity in gene expression profiles of soft tissue sarcomas. AB - Soft-tissue sarcomas (STSs) constitute more than 30 histologic entities. In addition, within each entity, tumors are often heterogeneous in macroscopic features, genetic alterations, microscopic appearance, and clinical course. Therefore, there has been concern about whether a single tumor sample can provide a gene expression profile representative of the entire tumor. We used 27-k cDNA microarray slides to assess the importance of intratumor versus intertumor heterogeneity of the gene expression profiles of 2 morphologically heterogeneous STSs. Multiple pieces of tumor (8 and 10 pieces) were obtained from a myxoid variant of malignant fibrous histiocytoma (MFH) and a leiomyosarcoma (LMS), respectively, and the expression patterns were compared with single tumor samples from 20 MFHs and 16 LMSs. Hierarchical clustering analysis of the expression profiles showed that samples from the same tumor clustered together. The average intratumor distance was considerably shorter than the average intertumor distance in both LMS and MFH. In addition, tumor subclusters that distinguished different macroscopic parts of the tumor could be discerned. We concluded that intratumor variability exists but that accurate gene expression profiling also could be obtained using single samples from a large STS. PMID- 15834946 TI - Arrested spread of vesicular stomatitis virus infections in vitro depends on interferon-mediated antiviral activity. AB - A quantitative understanding of the innate immune response will enable its recruitment against emerging, poorly characterized, or weaponized viral pathogens. To gain insights into how the innate responses can limit viral spread, we used quantitative focal infections to study how the spread of recombinant vesicular stomatitis viruses (VSV) on baby hamster kidney (BHK) and delayed brain tumor (DBT) cell monolayers is affected by innate cellular antiviral responses. We observed that rates of infection spread correlated with one-step growth rankings for four ectopic VSV strains: N1, N2, N3, and N4. However, this correlation was lost for M51R, a recombinant VSV mutant that lacks the ability to shut-off host gene expression. In BHK cells, M51R spread at two-thirds the rate of the recombinant control virus, XK3.1, even though their one-step growth was comparable. In DBT cells, M51R infections failed to spread beyond the site of inoculation. Addition of anti-interferon antibody restored M51R spread and one step growth to wild-type levels. Interestingly, the antibody enhanced the spread of wild-type virus but not its growth. These results suggest that while the rate of viral spread generally correlates with the rate of viral growth, the induction of cellular antiviral activities can be in some cases, the overriding factor in both spread and growth. In summary, focal infections enabled us to visualize and quantify how viral spread was inhibited by cellular antiviral activities. This study demonstrates a mechanism for quantifying how innate cellular responses can mitigate infection spread in vitro. PMID- 15834948 TI - Microfabricated grooved substrates as platforms for bioartificial liver reactors. AB - An extracorporeal bioartificial liver device has the potential to provide temporary hepatic support for patients with liver failure. Our goal was to optimize the flow environment for the cultured hepatocytes in a flat-plate bioreactor, specifically focusing on oxygen delivery using high medium flow rates while reducing the detrimental effects of the resulting shear stresses. We used photolithographic techniques to fabricate microgrooves onto the underlying glass substrate. The microgrooves, perpendicular to the axial flow direction, protected the hepatocytes from the shear stress induced by the flowing medium. Using finite element analysis, we found that the velocity gradient change near the cell surface (i.e., bottom of the grooves) was smaller than that near the top surface of the flow channel, indicating that the grooves would provide protection to the attached cells from the mechanical effects of the flowing medium. We also determined that the shear stress at the cell surface could be reduced by as much as 30 times (channel height of 100 microm) in the grooved-substrate (0.5 dyn/cm(2)) bioreactor compared to the flat-substrate (15 dyn/cm(2)) bioreactor for a medium flow rate of 4.0 mL/min. Albumin and urea synthesis rates of hepatocytes cocultured with 3T3-J2 fibroblasts remained stable over 5 days of perfusion in the grooved-substrate bioreactor, whereas in the flat-substrate bioreactor they decreased over the same time period. These studies indicate that under "high" flow conditions the microgrooved-substrate in the bioreactor can decrease the detrimental effects of shear stress on the hepatocytes while providing adequate oxygenation, thereby resulting in stable liver-specific function. PMID- 15834949 TI - On-line monitoring of bioreductions via membrane introduction mass spectrometry. AB - Real-time and on-line continuous monitoring of reactants, intermediates, and final products for dicarbonyl compound bioreduction in a continuous plug flow reactor packed with baker's yeast (Saccharomyces cerevisiae) whole cells immobilized on calcium alginate beads was performed by membrane introduction mass spectrometry (MIMS) via selective ion monitoring. PMID- 15834950 TI - Characterization of changes in PER.C6 cellular metabolism during growth and propagation of a replication-deficient adenovirus vector. AB - PER.C6 cells were cultivated for propagation of a replication-defective adenovirus vector in serum-free suspension bioreactors. Cellular metabolism during cell growth and adenovirus propagation was fully characterized using on line and off-line methods. The energy metabolism was found to accelerate transiently after adenovirus infection with increases in glucose and oxygen consumption rates. Similar to other mammalian cells, glucose utilization was highly inefficient and a high lactate:glucose yield was observed, both before and after virus infection. A higher consumption of most of the essential amino acids was observed transiently after the infection, likely due to increased protein synthesis requirements for virus propagation. To improve virus propagation, a medium exchange strategy was implemented to increase PER.C6 cell concentration for infection. During cell growth, a 50% increase in glucose consumption and lactate production rates was observed after initiation of the medium exchange in comparison to the batch phase. This decrease in medium capacity only affected the central carbon metabolism and no increase in amino acid consumption was observed. In addition, even though cell concentrations of up to 10 x 10(6) cells/mL were reproducibly obtained by medium exchange, infections at cell concentrations higher than 1 x 10(6) cells/mL did not proportionally improve volumetric adenovirus productivities. No measured nutrient limitation was observed at those high cell concentrations, indicating that adenovirus cell-specific productivity at higher cell concentrations is highly dependent on cell physiology. These results provide a better understanding of PER.C6 cellular metabolism and a basis for intensifying PER.C6 growth and adenovirus propagation. PMID- 15834951 TI - Oxidative stress in plant cell culture: a role in production of beta-thujaplicin by Cupresssus lusitanica suspension culture. AB - Oxidative stress is a common physiological stress that often challenges plants. Reactive oxygen species (ROS) are major factors in oxidative stress that significantly affect plant cell growth and secondary metabolism. Here we used beta-thujaplicin production by Cupressus lusitanica cell culture as an example to demonstrate the common occurrence of oxidative stress in cultivated plant cells and its effect on multiple aspects of cell culture process. C. lusitanica cells cultivated under Fe(2+) stress generate a significant level of ROS, and oxidative stress also occurs at late stages of C. lusitanica cell cultures under normal conditions. ROS production inhibited cell growth, induced lipid peroxidation and cell death, and enhanced ethylene and beta-thujaplicin production. It is demonstrated that Fe(2+) stress enhances ROS production via the Fenton reaction and promotes beta-thujaplicin production via ROS-induced lipid peroxidation that may activate cyclic oxylipin and ethylene pathways. Results further indicate that H(2)O(2) is a positive signal for beta-thujaplicin production, whereas superoxide anion radical (O(2) (- )) negatively affects beta-thujaplicin induction and strongly induces cell death. The study suggests that evaluating the oxidative stress and plant responses in a cell culture process is very necessary and important for understanding biochemical processes and for gaining the maximal productivity of target secondary metabolites. PMID- 15834953 TI - A base-pairing model of duplex formation. I. Watson-Crick pairing geometries. AB - We present a base-pairing model of oligonucleotide duplex formation and show in detail its equivalence to the nearest-neighbor dimer methods from fits to free energy of duplex formation data for short DNA-DNA and DNA-RNA hybrids containing only Watson-Crick pairs. For completeness, the corresponding RNA-RNA parameters are included. In this approach, the connection between rank-deficient polymer and rank-determinant oligonucleotide parameter sets for DNA duplexes is transparent. The method is generalized to include RNA-DNA hybrids where the rank-deficient model with 11 dimer parameters in fact provides slightly improved predictions relative to the standard method with 16 independent dimer parameters (DeltaG mean errors of 4.5 and 5.4%, respectively). PMID- 15834952 TI - Cultures of rat astrocytes challenged with a steady supply of glutamate: new model to study flux distribution in the glutamate-glutamine cycle. AB - Glutamate metabolism in astrocytes was studied using an experimental setup that simulates the role of neurons (glutamate producers and glutamine consumers) by the addition of glutaminase to the culture medium. Thereby, a steady supply of glutamate was imposed at the expense of glutamine, and the stress intensity was manipulated by changing the glutaminase concentration. Glutamate supply rates in the range 8-23 nmol/min/mg protein were examined for periods of up to 48 h. When the glutamate supply rate exceeded the uptake rate of this amino acid, a transient increase in the extracellular concentration of glutamate was observed. In response to this stress, the fluxes through the glutamate transporter and glutamine synthetase were increased considerably, and the extracellular concentration of glutamate was eventually restored to a low level. The increased levels of glutamine synthetase were demonstrated by immunoblotting analysis. The effect on glutamate metabolism of the transaminase inhibitor, aminooxyacetic acid (AOAA), and of NH4Cl was also investigated. The supply of glutamate caused a concomitant reduction in the levels of phosphocreatine, phosphoethanolamine, and phosphocholine without affecting the ATP pool. Glutamine synthetase was shown to be is a key element in the control of glutamate metabolism in astrocytic cultures. The metabolic fate of glutamate depends greatly on the time of endurance to the challenge: in naive cells, glutamate was primarily metabolized through the transaminase pathway, while in well-adapted cells glutamate was converted almost exclusively through glutamine synthetase. PMID- 15834955 TI - Expression patterns of the three Teashirt-related genes define specific boundaries in the developing and postnatal mouse forebrain. AB - We compare the expression patterns of the three mouse Teashirt (mTsh) genes during development of the forebrain and at a postnatal stage. During development, mTsh genes are expressed in domains that are restricted both dorsoventrally and rostrocaudally, with major changes in expression level coinciding with compartment boundaries. Striking complementarities in the distribution of mTsh transcripts were observed in the developing diencephalon, telencephalon, and olfactory bulb (OB). A mTsh1-positive cell population is part of the DLX-positive population localized in the dorsalmost portion of the lateral ganglionic eminence (dLGE). Comparison of the mTsh1 expression domain with the domains of Er81 and Islet1, which mark two distinct progenitor populations in the subventricular zone of the LGE, suggests that mTsh1 marks OB interneuron progenitors. Furthermore, the distinct expression patterns of mTsh1 and mTsh2 in the ventral LGE and the dLGE highlight the differential contributions of these structures to the striatum and the amydaloid complex. For Sey/Sey mutants, we show that Pax6 function is critical for the correct specification of the mTsh1+ population in the dLGE during embryogenesis. At postnatal stages in the OB, mTsh1 is expressed in granule and periglomerular cells, which originate from the subpallium during development. Furthermore, mTsh1+ cells line the walls of the anterior lateral ventricle, a region that gives rise to the interneurons that migrate in the rostral migratory streams and populate the OB postnatally. Our results suggest a role for mTsh genes in the establishment of regional identity and specification of cell types in the developing and adult forebrain. PMID- 15834956 TI - Withdrawal from progesterone increases expression of alpha4, beta1, and delta GABA(A) receptor subunits in neurons in the periaqueductal gray matter in female Wistar rats. AB - Premenstrual dysphoric disorder (PMDD) shows comorbidity with other psychiatric conditions such as panic disorder (PD). The symptoms of both conditions are exacerbated during the late luteal phase of the menstrual cycle, when progesterone levels fall sharply. The present study investigated the effect of withdrawal from progesterone (PWD) on expression of alpha4, beta1, and delta GABA(A) receptor subunits in neurons within the panic circuitry of the midbrain periaqueductal gray matter (PAG) in adult female Wistar rats. Immunostaining for alpha4, beta1, and delta GABA(A) receptor subunits was present in neurons throughout the PAG in vehicle-treated animals (VEH), in rats after 24 hours withdrawal from a progesterone dosing regime (PWD, 5 mg kg(-1) i.p. twice daily for 6 days), and in animals maintained on progesterone for 7 days (HP). Compared to HP and VEH animals, which did not differ significantly from each other, the number of immunostained neurons present in the PAG of PWD rats was significantly higher. The effect was most pronounced in the dorsolateral column of the PAG. The parallel changes in the three GABA(A) receptor subunits suggests that falling progesterone levels may be associated with expression of new receptors of the alpha4beta1delta subtype. This could lead to functional changes in GABAergic transmission within the PAG. We suggest that changes in GABA(A) receptor-mediated inhibitory tone in the PAG consequent to withdrawal from progesterone may contribute to the increased anxiety and susceptibility to panic seen during the late luteal phase of the menstrual cycle in PMDD and PD patients. PMID- 15834957 TI - Differential expression of CRMP1, CRMP2A, CRMP2B, and CRMP5 in axons or dendrites of distinct neurons in the mouse brain. AB - CRMP1, CRMP2, and CRMP5 have been identified as cytosolic proteins relaying semaphorin 3A signalling, one of the molecular cues conducting axon and dendrite growth and guidance. They are highly expressed during brain ontogenesis, but, because of their lower levels in the adult, their distribution in the mature brain is poorly documented. By using specific antibodies, we investigated the cellular distribution of these CRMPs in different adult brain structures and in neural cell cultures with a special focus on the splice variants CRMP2A and CRMP2B. In brain sections of adult mouse, CRMP1, CRMP2B, and CRMP5 were located predominantly in dendrites of specific neuronal populations, such as cortical pyramidal neurons, hippocampal CA1 pyramidal cells, or Purkinje cerebellar cells. On the contrary, CRMP2A was specifically associated with axons of the corpus callosum, bundles of the striatum, and mossy fibers of the hippocampus. In cultures of cortical neurons, CRMP1, CRMP2A, CRMP2B, and CRMP5 were equally distributed throughout cell bodies, axons, or dendrites of neurons, whereas CRMP2A and CRMP5 were completely absent from Purkinje cerebellar cells in 12-day old animals. By comparison, oligodendrocytes exclusively express CRMP2B and CRMP5 in cell bodies and processes both in situ in the adult brain and in primary cultures. Overall, our results demonstrate specific subcellular localizations of CRMP1, CRMP2A, CRMP2B, and CRMP5 depending on cell types, neuronal compartment, and developmental stage. This study suggests that, beyond their signalling function in axon outgrowth and guidance, CRMPs also play a role in mature neurons both in axons and in dendrites. PMID- 15834958 TI - Retrograde tracing of the subset of afferent connections in mouse barrel cortex provided by zincergic neurons. AB - The barrel cortex of rodents is densely innervated by a prominent subclass of glutamatergic neurons that sequester and release zinc from their synaptic boutons. These neurons may play an important role in barrel cortex function and plasticity, as zinc has been shown to modulate synaptic function by regulating neurotransmitter release, excitatory and inhibitory amino acid receptors, and second messenger signaling cascades. Here, we utilized intracortical infusions of sodium selenite to identify the source of the zincergic innervation to the mouse barrel cortex. Our results demonstrate that the majority of zincergic projections to the barrel cortex arose from ipsilateral and callosal neurons, situated in cortical layers 2/3 and 6. Regionally, these labeled neurons were most abundant within the barrel cortex itself, posterior parietal association cortex, secondary somatosensory cortex, and motor cortex. Labeled neurons were also found in other somatosensory regions corresponding to the trunk, fore- and hindlimb, as well as more distant regions such as the visual, rhinal, dorsal peduncular and insular cortices, the claustrum, and lateral and basolateral amygdaloid nuclei. Further, some mice were injected with the retrograde tracer cholera toxin subunit B to compare retrograde labeling of zincergic neurons with that of the general population of neurons innervating the barrel cortex. Our data indicate that all cortical regions providing inputs to the barrel cortex possess a zincergic component, whereas those from thalamic or brainstem structures do not. These findings demonstrate that zincergic pathways comprise a chemospecific associational network that reciprocally interconnects the barrel cortex with other cortical and limbic structures. PMID- 15834959 TI - Postnatal development of the cholinergic innervation in the dorsal hippocampus of rat: Quantitative light and electron microscopic immunocytochemical study. AB - Choline acetyltransferase (ChAT) immunocytochemistry was used to examine the distribution and ultrastructural features of the acetylcholine (ACh) innervation in the dorsal hippocampus of postnatal rat. The length of ChAT-immunostained axons was measured and the number of ChAT-immunostained varicosities counted, in each layer of CA1, CA3, and dentate gyrus, at postnatal ages P8, P16, and P32. At P8, an elaborate network of varicose ChAT-immunostained axons was already visible. At P16, the laminar distribution of this network resembled that in the adult, but adult densities were reached only by P32. Between P8 and P32, the mean densities for the three regions increased from 8.4 to 14 meters of axons and 2.3 to 5.7 million varicosities per cubic millimeter of tissue. At the three postnatal ages, the ultrastructural features of ChAT-immunostained axon varicosities from the strata pyramidale and radiatum of CA1 were similar between layers and comparable to those in adult, except for an increasing frequency of mitochondria (up to 41% at P32). The proportion of these profiles displaying a synaptic junction was equally low at all ages, indicating an average synaptic incidence of 7% for whole varicosities, as previously found in adult. The observed junctions were small, usually symmetrical, and made mostly with dendritic branches. These results demonstrate the precocious and rapid maturation of the hippocampal cholinergic innervation and reveal its largely asynaptic nature as soon as it is formed. They emphasize the remarkable growth capacities of individual ACh neurons and substantiate a role for diffuse transmission by ACh during hippocampal development. PMID- 15834962 TI - Afferent projections to pharynx and soft palate motoneurons: a light and electron microscopical tracing study in the cat. AB - Pharynx and soft palate are muscles for respiration, vocalization, swallowing, and vomiting. In cat, motoneurons innervating pharynx/soft palate are located in the dorsal group of the nucleus ambiguus (dgNA) in the medulla oblongata. In cat, dgNA is the only part of nucleus ambiguus that can be distinguished as a separate cell group, which makes it possible to study its afferent input. In two cats, WGA HRP injections in dgNA and surrounding tegmentum resulted in retrogradely labeled cells at several levels of the neuraxis. In 170 cases anterograde tracers were injected in areas in which the cells of origin were identified. Results demonstrate that dgNA afferents originate from the tegmentum dorsolateral to the superior olivary complex, medullary ventromedial tegmentum, caudal raphe nuclei, medullary lateral tegmental field, nucleus retroambiguus (NRA), and adjoining tegmentum, extending into the first cervical segment of the spinal cord. In order to determine whether periaqueductal gray (PAG) and parabrachial nuclei (PB) make synaptic contacts with dgNA, ultrastructural studies combined anterograde tracing from PAG, PB, and NRA with retrograde tracing of pharyngeal and soft palate motoneurons. The results showed that PB, but not PAG, projects to the dgNA and that NRA afferent synapses are three times as numerous as those from PB. The morphology of PB and NRA synapses is consistent with excitatory input. In conclusion, pharyngeal and soft palate motoneurons receive their afferents almost exclusively from the pontine and medullary tegmentum and first cervical spinal segment. PMID- 15834963 TI - Voluntary exercise alters the cytoarchitecture of the adult dentate gyrus by increasing cellular proliferation, dendritic complexity, and spine density. AB - Voluntary exercise produces a dramatic increase in the number of bromodeoxyuridine (BrdU)-positive cells in the adult dentate gyrus (DG); however, it has never been determined whether this increase reflects neurogenic activity or some exercise-induced change in the metabolic processing of systemically injected BrdU. In these experiments, we show that 1) 200 mg/kg is a saturating dose for single injections of BrdU in both control and voluntary exercise animals; 2) there is significantly more cell labeling in animals that exercise when saturating doses of BrdU are employed; 3) high doses of BrdU do not affect the number, appearance, or distribution of labeled cells; 4) voluntary exercise leads to similar increases in the number of cells expressing Ki67, an intrinsic marker of cellular proliferation; 5) both dendritic length and complexity are significantly increased in the DG of animals that exercise; and 6) spine density is significantly greater on dendrites in the DG following voluntary exercise. This study demonstrates that exercise up-regulates neurogenic activity in the DG of adult rats, independently of any putative changes in altered BrdU metabolism, and that it also substantially alters the morphology of dentate granule cell dendrites. The dramatic changes in the cytoarchitecture of the DG induced by voluntary exercise might underlie the enhancement of hippocampal long-term potentiation and hippocampal-dependent memory that our group has previously described. These results suggest that exercise may be an effective component of therapeutic regimes aimed at improving the functioning of individuals with neuropathologies that involve the degradation of cells in the hippocampus. PMID- 15834965 TI - The detection of modafinil and its major metabolite in equine urine by liquid chromatography/mass spectrometry. AB - A method has been developed for the detection of modafinil and its major metabolite, modafinil acid, in equine urine by solid-phase extraction and positive ion electrospray ionisation liquid chromatography/mass spectrometry. The method has been applied to the analysis of equine urine samples obtained after the oral administration of modafinil. Modafinil acid was the major component in the urine, and was detected up to 4 days post-administration. Unchanged modafinil was present at substantially lower concentrations, and was detected for only 24 hours. PMID- 15834966 TI - Family centred decision making and non-disclosure of diagnosis in a South East Asian oncology practice. AB - AIM: To audit the extent of non-disclosure of cancer diagnosis (NDD) in South East Asian patients referred to a Western trained oncologist, and determine the factors associated with this pattern of decision-making. METHODS: Over a 12-month period, all new patients referred to a single radiation oncologist's practice were prospectively audited at time of the initial consultation. Data obtained included patient demographic features, tumour details, proposed treatment and decision-making process. A shared decision-making consultation was attempted but if there was a persistent request by family to avoid disclosure, then a family centred model was adopted. Patient autonomy was maintained by formally asking desire to participate in decision-making process, and subsequent permission then sought to involve family members as surrogates. Treatment aim, predicted median survival and NDD was recorded at end of consultation. Endpoints of patient NDD and non-discussion of prognosis were audited over a 12-month period, and factors potentially associated with these endpoints were assessed. RESULTS: Over a 12 month period 369 new patients were referred to the oncologist's practice. Forty per cent were >65 years, 84% Chinese race, and 66% non-English speaking. Lung (33%) and breast (32%) primaries were prominent. Forty-seven per cent were managed with palliative intent and 24% had expected median survivals of <6 months. NDD was recorded in 66 patients or 17% of the patient group. Quantitative discussion of prognosis was avoided in 36.8% of patients. On univariate analysis advanced patient age (p<0.001, OR=9.6, 95% CI 4.9-18.9), female sex (p=0.035, OR=1.8, 95% CI 1.04-3.1), non-English speaking (p<0.001, OR=21.4, 95% CI 5.1 89.1), palliative treatment aim (p<0.001, OR=5.9, 95% CI 3.1-11.2) and short expected median survival (p<0.001, OR=4.0, 95% CI 2.3-7.1) were associated with NDD. Advanced patient age (p<0.001, OR=7.9, 95% CI 3.6-17.5), female sex (p<0.001, OR=6.4, 95% CI 2.8-14.7), non-English speaking (p=0.010, OR=7.4, 95% CI 1.6-33.3) and palliative treatment aim (p=0.010, OR=3.3, 95% CI 1.3-8.0) remained significantly associated with NDD on multivariate logistic regression analysis. CONCLUSIONS: A high rate of desired NDD is evident in this Asian oncology population when a family-centred model to medical decision making is used. This data confirms that NDD and the model of decision making remains a significant ethical issue. PMID- 15834967 TI - The global shortage of registered nurses. PMID- 15834969 TI - UK government to sponsor research into gel against HIV. PMID- 15834968 TI - Focal adhesion kinase as a potential target in arsenic toxicity. PMID- 15834970 TI - Are you ready for the next influenza pandemic? PMID- 15834971 TI - Saving mothers, saving children. PMID- 15834972 TI - HIV and hepatitis co-infection: a consensus for treatment? PMID- 15834973 TI - Defining the concept of public information. PMID- 15834974 TI - Defining the concept of public information. PMID- 15834975 TI - Defining the concept of public information. PMID- 15834976 TI - Comparison of short-term treatments for GERD. PMID- 15834977 TI - AIDS pioneer advocates for therapeutic vaccine. PMID- 15834978 TI - Tell and kiss: dating and disclosure. PMID- 15834979 TI - "By the way, I'm poz". One man's experience with dating and disclosure. PMID- 15834980 TI - The disclosure dilemma. "I'm afraid my friends and family will be judgemental because my boyfriend has HIV". PMID- 15834981 TI - Mixed HIV status couples. Moving from research to practice. PMID- 15834982 TI - [Mendelian randomization. Danish Society of Epidemiology]. PMID- 15834983 TI - Tickborne meningoencephalitis, first case after 19 years in northeastern Germany. PMID- 15834984 TI - Social impact of leishmaniasis, Afghanistan. PMID- 15834985 TI - Methicillin-resistant Staphylococcus aureus toxic shock syndrome. PMID- 15834986 TI - Are SARS superspreaders cloud adults? PMID- 15834987 TI - Route of infection in melioidosis. PMID- 15834988 TI - Bordetella pertussis in adult pneumonia patients. PMID- 15834989 TI - SARS risk perception and preventive measures, Singapore and Japan. PMID- 15834990 TI - Fearsome creatures and nature's gothic. PMID- 15834991 TI - [Experience with a large-scale day care at a clinic]. PMID- 15834992 TI - Living in space: scientific, medical and cultural implications. A selection of papers presented at the 14th IAA Humans in Space Symposium. PMID- 15834993 TI - The USCCB and rape protocols. PMID- 15834994 TI - Papal address on food and water. PMID- 15834995 TI - John Paul II on the "vegetative state". PMID- 15834996 TI - Communicating bioastronautics research to students, families and the nation. AB - The National Space Biomedical Research Institute (NSBRI) is supporting the National Aeronautics and Space Administration's (NASA) education mission through a comprehensive Education and Public Outreach Program (EPOP) that communicates the excitement and significance of space biology to schools, families, and lay audiences. The EPOP is comprised of eight academic institutions: Baylor College of Medicine, Massachusetts Institute of Technology, Morehouse School of Medicine, Mount Sinai School of Medicine, Texas A&M University, University of Texas Medical Branch Galveston, Rice University, and the University of Washington. This paper describes the programs and products created by the EPOP to promote space life science education in schools and among the general public. To date, these activities have reached thousands of teachers and students around the US and have been rated very highly. PMID- 15834997 TI - Increasing student learning through space life sciences education. AB - Scientists and educators at Baylor College of Medicine are using space life sciences research areas as themes for middle school science and health instructional materials. This paper discusses study findings of the most recent unit, Food and Fitness, which teaches concepts related to energy and nutrition through guided inquiry. Results of a field test involving more than 750 students are reported. Use of the teaching materials resulted in significant knowledge gains by students as measured on a pre/post assessment administered by teachers. In addition, an analysis of the time spent by each teacher on each activity suggested that it is preferable to conduct all of the activities in the unit with students rather than allocating the same total amount of time on just a subset of the activities. PMID- 15834998 TI - The Unborn Victims of Violence Act. PMID- 15834999 TI - Recovery of lower limb function following 6 weeks of non-weight bearing. AB - Skeletal muscle weakness and atrophy occur following an extended period of decreased use, including space flight and limb unloading. It is also likely that affected muscles will be susceptible to a re-loading injury when they begin return to earth or weight bearing. However, there is a paucity of literature evaluating the response of human unloaded muscle to exercise and return to activity. The purpose of this pilot study was to evaluate the soreness, function and strength response of muscle to re-loading in seven patients who were non weight bearing for 6 weeks, compared to five healthy subjects. Function improved significantly over time for the patients but was still less than the healthy subjects over 12 weeks of physiotherapy. Concentric quadriceps muscle strength increased significantly over time for the patients. There was considerable variability in the patients' reports of muscle soreness but there were no significant changes over time or between groups. PMID- 15835000 TI - [Small-scale day care service at a clinic]. PMID- 15835002 TI - The pope moves backward on terminal case. PMID- 15835001 TI - On advocating infant euthanasia. PMID- 15835003 TI - The harm that religion does. PMID- 15835004 TI - From regenerative medicine to human design: what are we really afraid of? PMID- 15835005 TI - Humanism for personhood: against human-racism: a new fight for human rights. PMID- 15835007 TI - [Experience at a mini-day care at a clinic]. PMID- 15835006 TI - Spatial factors and muscle spindle input influence the generation of neuromuscular responses to stimulation of the human foot. AB - Removal of the mechanical pressure gradient on the soles leads to physiological adaptations that ultimately result in neuromotor degradation during spaceflight. We propose that mechanical stimulation of the soles serves to partially restore the afference associated with bipedal loading and assists in attenuating the negative neuromotor consequences of spaceflight. A dynamic foot stimulus device was used to stimulate the soles in a variety of conditions with different stimulation locations, stimulation patterns and muscle spindle input. Surface electromyography revealed the lateral side of the sole elicited the greatest neuromuscular response in ankle musculature, followed by the medial side, then the heel. These responses were modified by preceding stimulation. Neuromuscular responses were also influenced by the level of muscle spindle input. These results provide important information that can be used to guide the development of a "passive" countermeasure that relies on sole stimulation and can supplement existing exercise protocols during spaceflight. PMID- 15835008 TI - Reflections on genetic manipulation and duties to posterity: an engagement with Skene and Coady. AB - In addressing the regulation of human genetic futures, scientific standards concerning human kinds are endorsed by philosophical approaches that tend to exclude many people with genetic conditions from the deliberative process. In broadening the axiological, ontological and epistemological framework to include disability perspectives, the focus is shifted from questions of regulation to practical matters of participation, invoking ideals of community equality and enabled choice. In developing practical community engagements to deliberate upon genetic futures, a process that allows dialectical encounter between eugenic and non-eugenic approaches is envisioned so that strong versions of eugenics are avoided. PMID- 15835009 TI - Response to the Nancy Crick case. PMID- 15835010 TI - [The current status and future problems of psychiatric day care service in Japan]. PMID- 15835011 TI - Beyond Nancy Crick. PMID- 15835012 TI - Ethics, death and silence: a comment on the euthanasia debate. PMID- 15835013 TI - Quantification of spatial structure of human proximal tibial bone biopsies using 3D measures of complexity. AB - Changes in trabecular bone composition during development of osteoporosis are used as a model for bone loss in microgravity conditions during a space flight. Symbolic dynamics and measures of complexity are proposed and applied to assess quantitatively the structural composition of bone tissue from 3D data sets of human tibia bone biopsies acquired by a micro-CT scanner. In order to justify the newly proposed approach, the measures of complexity of the bone architecture were compared with the results of traditional 2D bone histomorphometry. The proposed technique is able to quantify the structural loss of the bone tissue and may help to diagnose and to monitor changes in bone structure of patients on Earth as well as of the space-flying personnel. PMID- 15835014 TI - Ethical regulation or regulating ethics? The need for both internal and external governance of human experimentation. AB - Research regulation is a timely topic for discussions in bioethics and public health policy. This response to articles in the previous special issue of the Monash Bioethics Review emphasises the importance of having both internal and external controls on human experimentation. Unless both elements are incorporated into research ethics governance frameworks, they will ultimately fail to achieve what should be their primary goal: human subject protection. PMID- 15835015 TI - Influence of metabolic control on the blood-retinal barrier in streptozocin diabetic rats. AB - One hundred and eight male Sprague-Dawley rats weighing 160-225 g were examined with vitreous fluorophotometry (VF). Fifty-five rats were made diabetic with streptozocin (43 with 65 mg/kg and 12 with 90 mg/kg); 53 nondiabetic rats served as controls. Fluorescein (7 mg/kg) was injected IV and VF was performed 60 min later. The diabetic rats had an abnormal blood-retinal barrier (BRB) after 4-7 days duration. The difference between the diabetic and non-diabetic rats was persistent up to 35 days (P < 0.02-0.001). The diabetic rats treated with 90 mg/kg streptozotocin had a more severe diabetes and a higher treated with 65 mg/kg (P < 0.05). The BRB function was normalized in 18 rats treated for 6-7 days with 5-6 IU long-acting insulin (P < 0.02). The results imply that streptozocin diabetic rats have an abnormal BRB and that this abnormality seems to be related to the diabetic state and to be reversible after insulin treatment. PMID- 15835017 TI - [Science of subjective experiences (awareness)--its starting point and outcome]. PMID- 15835016 TI - The ethics of germ line gene manipulation--a five dimensional debate. AB - Contributors to the debate surrounding the ethics of germ line gene manipulation have by and large concentrated their efforts on discussions of the potential risks that are associated with the use of this technology. Many international advisory committees have ruled out the acceptability of germ line gene manipulation at least for the time being. The purpose of this work is to generate much needed discussion on the many other ethical issues concerning the implementation of not only germ line gene manipulation but also other related biotechnologies. In this paper I systematically investigate and analyse the most salient issues put forward by proponents and opponents alike. I argue that if germ line manipulation proves to be a safe and effective procedure, then the principle of beneficence imposes on the medical profession a moral duty to pursue the technology. PMID- 15835018 TI - Thoracic sonography for pneumothorax: the clinical evaluation of an operational space medicine spin-off. AB - The recent interest in the use of ultrasound (US) to detect pneumothoraces after acute trauma in North America was initially driven by an operational space medicine concern. Astronauts aboard the International Space Station (ISS) are at risk for pneumothoraces, and US is the only potential medical imaging available. Pneumothoraces are common following trauma, and are a preventable cause of death, as most are treatable with relatively simple interventions. While pneumothoraces are optimally diagnosed clinically, they are more often inapparent even on supine chest radiographs (CXR) with recent series reporting a greater than 50% rate of occult pneumothoraces. In the course of basic scientific investigations in a conventional and parabolic flight laboratory, investigators familiarized themselves with the sonographic features of both pneumothoraces and normal pulmonary ventilation. By examining the visceral-parietal pleural interface (VPPI) with US, investigators became confident in diagnosing pneumothoraces. This knowledge was subsequently translated into practice at an American and a Canadian trauma center. The sonographic examination was found to be more accurate and sensitive than CXR (US 96% and 100% versus US 74% and 36%) in specific circumstances. Initial studies have also suggested that detecting the US features of pleural pulmonary ventilation in the left lung field may offer the ability to exclude serious endotracheal tube malpositions such as right mainstem and esophageal intubations. Applied thoracic US is an example of a clinically useful space medicine spin-off that is improving health care on earth. PMID- 15835020 TI - [Studies on the intermediate course of schizophrenia]. PMID- 15835019 TI - Virtue ethics and public health: a practice-based analysis. AB - Public health plays an important, albeit often unnoticed, role in protecting and promoting the health of populations. The activities of public health are complex, performed by multiple professionals, and range from the innocuous to the intrusive. Ethical analyses in public health reflect some of this complexity and fragmentation, with no one approach able to capture the full range of ethical considerations raised by public health activities. There are however, good reasons why we should pursue such analyses. Providing a robust ethical framework for public health may promote the identity and function of public health, address some of the shortcomings of utilitarianism, and help to combat the threat that public health faces through lack of political will in many parts of the world. In this paper I argue that Alasdair MacIntyre's account of practices and virtues can make a valuable contribution to public health ethics. The first part of the paper argues that public health may properly be described as the type of practice that provides an arena for the exercise of virtues. This is followed by an analysis of the three virtues of honesty, courage and justice in public health practice. Using virtue theory captures morally important elements of public health and helps to maintain awareness of significant moral values in the practice of public health. Such awareness is crucial in maintaining and defending the integrity of public health. PMID- 15835021 TI - Rethinking the 'right not to know'. AB - The idea that an individual has a 'right not to know' genetic information about himself or herself is entrenched in both the policy sphere and the genetic counselling ethos. In this paper, I interrogate this idea of a 'right not to know', questioning particularly its status as a right. I identify the conception of rights that seems to underlie the posited 'right not to know' as a conception of rights in which they are prioritised non-outweighable interests. Turning to a series of hypothetical situations in which the 'right not to know' is invoked, I argue that the 'right not to know' is not a right consistent with such a conception. The 'right not to know' can only exist in a framework of rights in which they lack significant moral weight. Thus, considering the gravity of rights in popular understanding, I suggest caution in continuing use of the phrase 'right not to know' in the context of genetic testing. This investigation suggests that structuring the debate around genetic ignorance in terms of the 'right not to know' is not conducive to moving forward ethical thinking in this area. PMID- 15835022 TI - [Water intoxication]. PMID- 15835023 TI - Microbiological status of cosmonauts during orbital spaceflights on Salyut and Mir orbital stations. AB - The main feature of the human-microorganism eco-system forming in the environment of manned space vehicles is the periodic accumulation of the potential for pathogenicity. This process is characterized by the activation of opportunistic pathogens, representatives of which grow in large number within the system, settle in various niches, and demonstrate expansion, intruding into microbiocenoses of open biotopes of humans who, until contamination, have been clean of these microorganisms. Clones (similar to hospital strains) arising out of these populations exhibit the ability to spread "epidemically" within isolated teams. PMID- 15835024 TI - Access to essential drugs, human rights and global justice. PMID- 15835025 TI - Should third party consent to research be mandated? Should there be a right for third parties to have data about them withdrawn from a research project? Two perspectives. PMID- 15835026 TI - The legacy and the future. 30 years after the Belmont Report, Beauchamp sets the record straight. PMID- 15835027 TI - California adolescents increasingly inactive. PMID- 15835028 TI - Too many California adults are tipping the scales at an unhealthy weight. PMID- 15835029 TI - Humans to Mars: a feasibility and cost-benefit analysis. AB - Mars is a compelling astrobiological target, and a human mission would provide an opportunity to collect immense amounts of scientific data. Exploration alone, however, cannot justify the increased risk. Instead, three factors drive a human mission: economics, education, and exploration. A human mission has a unique potential to inspire the next generation of young people to enter critically needed science and engineering disciplines. A mission is economically feasible, and the research and development program put in place for a human mission would propel growth in related high-technology industries. The main hurdles are human physiological responses to 1-2 years of radiation and microgravity exposure. However, enabling technologies are sufficiently mature in these areas that they can be developed within a few decade timescale. Hence, the decision of whether or not to undertake a human mission to Mars is a political decision, and thus, educational and economic benefits are the crucial factors. PMID- 15835030 TI - [A 90-year-old woman with acute abdomen]. AB - Bowel strangulation is defined as mechanical obstruction associated with compromised blood circulation of the bowel wall. Delayed diagnosis and treatment are the main causes of the high mortality. A 90-year-old woman was admitted to our hospital with acute abdominal pain. Plain abdominal x-ray studies were inconclusive. However, bowel strangulation was suggested by computer tomography and the diagnosis was confirmed under the laparotomy. In the case of bowel strangulation, abdominal CT is a fast, reliable and useful diagnostic tool when imaging is considered necessary before a laparotomy. PMID- 15835031 TI - [Use of computer tomography in acute abdomen]. PMID- 15835032 TI - Artificial gravity--head movements during short-radius centrifugation: influence of cognitive effects. AB - Short-radius centrifugation is a potential countermeasure against the effects of prolonged weightlessness. Head movements in a rotating environment, however, induce serious side effects: inappropriate vestibular ocular reflexes (VOR), body tilt illusions and motion sickness induced by cross-coupled accelerations on a rotating platform. These are well predicted by a semicircular canal model. The present study investigates cognitive effects on the inappropriate VOR and the illusory sensations experienced by subjects rotating on a short-radius centrifuge (SRC). Subjects (N=19) were placed supine on a rotating horizontal bed with their head at the center of rotation. To investigate the extent to which they could control their sensations voluntarily, subjects were asked alternatively to "fight" (i.e. to try to resist and suppress) those sensations, or to "go" with (i.e. try to enhance or, at least, acquiesce in) them. The only significant effect on the VOR of this cognitive intervention was to diminish the time constant characterizing the decay of the nystagmus in subjects who had performed the "go" (rather than the "fight") trials. However, illusory sensations, as measured by reported subjective intensities, were significantly less intense during the "fight" than during the "go" trials. These measurements also verified an asymmetry in illusory sensation known from earlier experiments: the illusory sensations are greater when the head is rotated from right ear down (RED) to nose up (NU) posture than from NU to RED. The subjects habituated, modestly, to the rotation between their first and second sequences of trials, but showed no better (or worse) suppression of illusory sensations thereafter. No significant difference in habituation was observed between the "fight" and "go" trials. PMID- 15835033 TI - Artificial gravity: a possible countermeasure for post-flight orthostatic intolerance. AB - Four payload crewmembers were exposed to sustained linear acceleration in a centrifuge during the Neurolab (STS-90) flight. In contrast to previous studies, otolith-ocular reflexes were preserved during and after flight. This raised the possibility that artificial gravity may have acted as a countermeasure to the deconditioning of otolith-ocular reflexes. None of the astronauts who were centrifuged had orthostatic intolerance when tested with head-up passive tilt after flight. Thus, centrifugation may also have helped maintain post-flight hemodynamic responses to orthostasis by preserving the gain of the otolith sympathetic reflex. A comparison with two fellow Neurolab orbiter crewmembers not exposed to artificial gravity provided some support for this hypothesis. One of the two had hemodynamic changes in response to post-flight tilt similar to orthostatically intolerant subjects from previous missions. More data is necessary to evaluate this hypothesis, but if it were proven correct, in-flight short-radius centrifugation may help counteract orthostatic intolerance after space flight. PMID- 15835034 TI - Ethical issues in therapy: therapist self-disclosure of sexual feelings. AB - Although therapist sexual attraction to clients is common, and therapist self disclosure is an often-used intervention, therapist self-disclosure of sexual feelings to clients is an understudied phenomenon. In this article, I critically review the small base of literature on therapist self-disclosure of sexual feelings, including information on prevalence rates, empirical research, and case studies. By incorporating these findings with information from relevant sections of the American Psychological Association (2002) Ethics Code, my intent is to evaluate different aspects of therapist self-disclosure of sexual feelings and arrive at conclusions regarding therapists' use of these disclosures. It appears that direct, explicit disclosure of sexual feelings can run the risk of harming clients and may therefore be unethical. Therefore, the use of this technique is discouraged. I discuss the issue of using less explicit interventions. PMID- 15835035 TI - Lessons learned from Mir--a payload perspective. AB - Among the principal objectives of the Phase 1 NASA/Mir program were for the United States to gain experience working with an international partner, to gain working experience in long-duration space flight, and to gain working experience in planning for and executing research on a long-duration space platform. The Phase 1 program was to provide the US early experience prior to the construction and operation of the International Space Station (Phase 2 and 3). While it can be argued that Mir and ISS are different platforms and that programmatically Phase 1 and ISS are organized differently, it is also clear that many aspects of operating a long-duration research program are platform independent. This can be demonstrated by a review of lessons learned from Skylab, a US space station program of the mid-1970s, many of which were again "learned" on Mir and are being "learned" on ISS. Among these are optimum crew training strategies, on-orbit crew operations, ground support, medical operations and crew psychological support, and safety certification processes. PMID- 15835036 TI - Ethical considerations in the use of nonerotic touch in psychotherapy with children. AB - Although touch frequently occurs in psychotherapy with children, there is little written on the ethical considerations of therapeutic touch. Because physical contact does occur, therapists must consider if, how, and when it is used, for both their clients' safety and their own. In this review, I further develop the issues suggested by Aquino and Lee (2000) in the use of nurturing touch in therapy by considering many types of touch that occur in psychotherapy with children; the possible positive role of touch; clients' perception of touch in therapy; considerations related to the therapist, the child's safety, and any history of abuse in the child's and family's background; and other practical considerations. I list guidelines. PMID- 15835037 TI - Accomplishments in bioastronautics research aboard International Space Station. AB - The tenth long-duration expedition crew is currently in residence aboard International Space Station (ISS), continuing a permanent human presence in space that began in October 2000. During that time, expedition crews have been operators and subjects for 18 Human Life Sciences investigations, to gain a better understanding of the effects of long-duration spaceflight on the crewmembers and of the environment in which they live. Investigations have been conducted to study: the radiation environment in the station as well as during extravehicular activity (EVA); bone demineralization and muscle deconditioning; changes in neuromuscular reflexes; muscle forces and postflight mobility; causes and possible treatment of postflight orthostatic intolerance; risk of developing kidney stones; changes in pulmonary function caused by long-duration flight as well as EVA; crew and crew-ground interactions; changes in immune function, and evaluation of imaging techniques. The experiment mix has included some conducted in flight aboard ISS as well as several which collected data only pre- and postflight. The conduct of these investigations has been facilitated by the Human Research Facility (HRF). HRF Rack 1 became the first research rack on ISS when it was installed in the US laboratory module Destiny in March 2001. The rack provides a core set of experiment hardware to support investigations, as well as power, data and commanding capability, and stowage. The second HRF rack, to complement the first with additional hardware and stowage capability, will be launched once Shuttle flights resume. Future years will see additional capability to conduct human research on ISS as International Partner modules and facility racks are added to ISS. Crew availability, both as a subject count and time, will remain a major challenge to maximizing the science return from the bioastronautics research program. PMID- 15835038 TI - Ethics in violence against women research: the sensitive, the dangerous, and the overlooked. AB - Traditional disciplinary guidelines are inadequate to address some of the ethical dilemmas that emerge when conducting research on violence against women and girls. This article is organized according to the ethical principles of respect for persons, privacy and confidentiality, justice, beneficence, and nonmaleficence. In the article, I describe dilemmas involved in cross-cultural research, research on children, informed consent, voluntariness, coercion, deception, safety, mandated reporting, and dissemination. In the article, I include examples from qualitative and quantitative studies in many nations. I also offer suggestions for researchers and institutional review boards. PMID- 15835040 TI - Reflections on the researcher-participant relationship and the ethics of dialogue. AB - Research concerned with human beings is always an interference of some kind, thus posing ethical dilemmas that need justification of procedures and methodologies. It is especially true in social work when facing mostly sensitive populations and sensitive issues. In the process of conducting a research on the emotional life histories of Israeli men who batter their partners, some serious ethical questions were evoked such as (a) Did the participants really give their consent? (b) What are the limits of the researcher-participants relationship and who decides them? (c) For whom is the study beneficial? and (d) To what degree did the methodology fit with the participants? In this article, I discuss the Socratic idea of truth revealed through dialogue and the idea of reciprocity that was developed in Buber's (1949) ethics of dialogue and Habermas' (1990) communicative ethics. The 3 essential conclusions drawn from the ethical questions raised and the discussion of the thinkers that are mentioned previously are (a) dialogical methodology is ethically justified; (b) dynamic interactions give a more holistic perspective of the human nature, thus enriching the field; and (c) through dialogical methodology both researcher and participant profit from growth of knowledge, which is a key for empowerment and change. PMID- 15835039 TI - Neutral buoyancy and sleep-deprived serum factors alter expression of cytokines regulating osteogenesis. AB - We examined expression of genes associated with cytokine production, and genes implicated in regulating bone metabolism, in bone stromal and osteoblast cells incubated under standard ground conditions and under conditions of neutral buoyancy, and in the presence/absence of serum from normal or sleep-deprived mice. We observed a clear interaction between these two conditions (exposure to neutral buoyancy and serum stimulation) in promoting enhanced osteoclastogenesis. Both conditions independently altered expression of a number of cytokines implicated in the regulation of bone metabolism. However, using stromal cells from IL-1 and TNFx cytokine(r) KO mice, we concluded that the increased bone loss under microgravity conditions was not primarily cytokine mediated. PMID- 15835041 TI - Evaluation of theories of complex movement planning in different levels of gravity. AB - Due to high redundancy of degrees of freedom in the human body, we can perform any movement, from the simplest to the most complex, in many different ways. Several studies are still trying to identify the motor strategies that master this redundancy and generate the movements whose characteristics are highly stereotyped. The aim of this work is to build a simulator that is able to evaluate different motor planning hypotheses. The most interesting applications of this tool occur in studies of the motor strategy in microgravity conditions. The comparison between simulated movements and kinematics data recorded both on Earth, and during a 5-month mission on board the Mir station shows that for a complex whole-body movement (such as trunk bending) a single planning criterion cannot explain all movement aspects. However, the simulator allows an understanding of the motor planning adaptation of astronauts. In space, the lack of equilibrium constraint (which on Earth brings about the center of mass control) leads to a new motor strategy that minimizes dynamic interactions with the floor. PMID- 15835042 TI - Influence of the gravitational vertical on geometric visual illusions. AB - The occurrence of geometric orientation illusions and the perception of ambiguous figures were analyzed in 24 subjects during static body tilt relative to gravity on Earth. Results showed that illusions such as the Rock's diamond/square, the Ponzo illusion, and orientation contrast illusions occurred less frequently, and that depth reversal of ambiguous figures took more time when subjects were lying on their side or supine compared to upright, thus suggesting that the gravitational reference plays a significant role in these "visual" illusions. The structure of images, our representation of the environment, and orientation relative to gravity are all integral parts in interpreting visual images. In a weightless environment where no gravitational reference can be used, it is expected that similar alterations in visual perception will occur. PMID- 15835043 TI - Practical ethics. Cancer can't stop Betty. PMID- 15835044 TI - How does clients' method of payment influence psychologists' diagnostic decisions? AB - To what extent does payment method (managed care vs. out of pocket) influence the likelihood that an independent practitioner will assign a Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association, 1994) diagnosis to a client? When a practitioner does diagnose, how does payment method influence the specific choice of a diagnostic category? Independent practitioners responded to a vignette describing a fictitious client with symptoms of depression or anxiety. In half of the vignettes, the fictitious client intended to pay via managed care; in the other half, the fictitious client intended to pay out of pocket. Payment method had a very significant impact on diagnosis such that relative to out-of-pocket clients, managed care clients were much more likely to receive diagnoses and more likely to receive adjustment disorder diagnoses in particular. We discuss implications involving informed consent and other ethical issues. PMID- 15835045 TI - Induction of vascular endothelial phenotype and cellular proliferation from human cord blood stem cells cultured in simulated microgravity. AB - Recent studies have demonstrated that stem cells derived from adult hematopoietic tissues are capable of trans-differentiation into non-hematopoietic cells, and that the culture in microgravity (microg) may modulate the proliferation and differentiation. We investigated the application of microg to human umbilical cord blood stem cells (CBSC) in the induction of vascular endothelial phenotype expression and cellular proliferation. CD34+ mononuclear cells were isolated from waste human umbilical cord blood samples and cultured in simulated microg for 14 days. The cells were seeded in rotary wall vessels (RWV) with or without microcarrier beads (MCB) and vascular endothelial growth factor was added during culture. Controls consisted of culture in 1 G. The cell cultures in RWV were examined by inverted microscopy. Cell counts, endothelial cell and leukocyte markers performed by flow cytometry and FACS scan were assayed at days 1, 4, 7 and at the termination of the experiments. Culture in RWV revealed significantly increased cellular proliferation with three-dimensional (3D) tissue-like aggregates. At day 4, CD34+ cells cultured in RWV bioreactor without MCB developed vascular tubular assemblies and exhibited endothelial phenotypic markers. These data suggest that CD34+ human umbilical cord blood progenitors are capable of trans-differentiation into vascular endothelial cell phenotype and assemble into 3D tissue structures. Culture of CBSC in simulated microg may be potentially beneficial in the fields of stem cell biology and somatic cell therapy. PMID- 15835046 TI - Management. Lights, camera...action! PMID- 15835047 TI - Correction to "Empirical support for the United States Supreme Court's protection of the psychotherapist-patient privilege". PMID- 15835048 TI - Gender differences in leader and follower perceptions of social support in Antarctica. AB - The purpose of this study was to methodologically explore the links among social support, gender, age, prior experience, leader/follower status, and leadership effectiveness noted in previous accounts from Antarctic stations. Data for this study were collected from volunteers involved in Australian National Antarctic Research Expeditions conducted from 1996 to 2001. Multilevel analysis revealed that most of the variance in perceptions of social support was at the individual level (71%). Perceptions of social support had less variance at the group level (29%) and little variance at the weekly level. At the group level, the explanatory variables we examined included leadership effectiveness, gender similarity, and age similarity. At the individual level, the explanatory variables we examined included age, gender, prior experience, and leader/follower status. An interaction between gender and leader/follower status contributed to a significant model of variation in perceptions of social support. PMID- 15835051 TI - Leadership issues with multicultural crews on the international space station: lessons learned from Shuttle/Mir. AB - In isolated and confined environments, two important leadership roles have been identified: the task/instrumental role (which focuses on work goals and operational needs), and the supportive/expressive role (which focuses on morale goals and emotional needs). On the International Space Station, the mission commander should be familiar with both of these aspects of leadership. In previous research involving a 135-day Mir space station simulation in Moscow and a series of on-orbit Mir space station missions during the Shuttle/Mir program, both these leadership roles were studied. In new analyses of the Shuttle/Mir data, we found that for crewmembers, the supportive role of the commander (but not the task role) related positively with crew cohesion. For mission control personnel on the ground, both the task and supportive roles of their leader were related positively to mission control cohesion. The implications of these findings are discussed in terms of leadership on board the International Space Station. PMID- 15835052 TI - Distributed communication and psychosocial performance in simulated space dwelling groups. AB - The present report describes the development and application of a distributed interactive multi-person simulation in a computer-generated planetary environment as an experimental test bed for modeling the human performance effects of variations in the types of communication modes available, and in the types of stress and incentive conditions underlying the completion of mission goals. The results demonstrated a high degree of interchangeability between communication modes(audio, text) when one mode was not available. Additionally, the addition of time pressure stress to complete tasks resulted in a reduction in performance effectiveness, and these performance reductions were ameliorated via the introduction of positive incentives contingent upon improved performances. The results obtained confirmed that cooperative and productive psychosocial interactions can be maintained between individually isolated and dispersed members of simulated spaceflight crews communicating and problem-solving effectively over extended time intervals without the benefit of one another's physical presence. PMID- 15835053 TI - Bubble detector characterization for space radiation. AB - In light of the importance of the neutron contribution to the dose equivalent received by space workers in the near-Earth radiation environment, there is an increasing need for a personal dosimeter that is passive in nature and able to respond to this neutron field in real time. Recent Canadian technology has led to the development of a bubble detector, which is sensitive to neutrons, but insensitive to low linear energy transfer (LET) radiation. By changing the composition of the bubble detector fluid (or "superheat"), the detectors can be fabricated to respond to different types of radiation. This paper describes a preliminary ground-based research effort to better characterize the bubble detectors of different compositions at various charged-particle accelerator facilities, which are capable of simulating the space radiation field. PMID- 15835054 TI - Prediction of solar particle event proton doses using early dose rate measurements. AB - A methodology for predicting solar particle event doses using Bayesian inference is being developed. As part of this development, we have tested criteria for categorization of new solar particle events (SPE) using calculated asymptotic doses and dose rates for the 22 SPEs that occurred in 2001. In 9 out of 22 events, our criteria for categorization would have over-predicted the range of asymptotic doses in which the tested events would have fallen. In two cases, our methodology under-predicted the dose range in which the event would have fallen. In order to better predict a new event's group category and thus, to better restrict the Bayesian inference predictive model parameter space, we have reexamined our dose rate criteria for categorization of new events. We report the updating of the grouping criteria using data from the 22 SPEs of 2001, as well as five additional SPEs. Using the revised grouping criteria, we present an analysis of group categorization prediction results for the first ten SPEs of 2002. PMID- 15835055 TI - Interplanetary crew dose estimates for worst case solar particle events based on historical data for the Carrington flare of 1859. AB - Over the past two decades, hypothetical models of "worst-case" solar particle event (SPE) spectra have been proposed in order to place an upper bound on radiation doses to critical body organs of interplanetary crews on deep space missions. These event spectra are usually formulated using hypothetical extrapolations of space measurements for previous large events. Here we take a different approach. Recently reported analyses of ice core samples indicate that the Carrington flare of 1859 is the largest event observed in the past 500 years. These ice core data yield estimates of the proton fluence for energies greater than 30 MeV, but provide no other spectrum information. Assuming that the proton energy distribution for such an event is similar to that measured for other recent, large events, interplanetary crew doses are estimated for these hypothetical worst case SPE spectra. These estimated doses are life threatening unless substantial shielding is provided. PMID- 15835057 TI - Engaging space: extraterrestrial architecture and the human psyche. AB - The human fascination with exploring and inhabiting the space that lies beyond Earth's atmosphere continues to grow. Nevertheless, 40 years of experience to date have clearly established that humans in outer space routinely suffer significant psychological impairment arising from their stressful extraterrestrial living conditions. This paper explores those extraterrestrial conditions through the interactions between the extraordinarily harsh environment of outer space, the sensations that humans encounter in space, and the qualities of a habitat that physically interposes itself between the two. The objective of this paper is to develop a habitat that expresses the extraterrestrial condition while supporting the mental health of its inhabitants, so as to augment the success of prolonged extraterrestrial residence and interplanetary travel. PMID- 15835056 TI - A Canadian high-energy neutron spectrometry system for measurements in space. AB - Bubble Technology Industries Inc. (BTI), with the support of the Canadian Space Agency, has finished the construction of the Canadian High-Energy Neutron Spectrometry System (CHENSS). This spectrometer is intended to measure the high energy neutron spectrum (approximately 1-100 MeV) encountered in spacecraft in low earth orbit. CHENSS is designed to fly aboard a US space shuttle and its scientific results should facilitate the prediction of neutron dose to astronauts in space from readings of different types of radiation dosimeters that are being used in various missions. PMID- 15835058 TI - Analysis and modeling of information flow and distributed expertise in space related operations. AB - Evolving space operations requirements and mission planning for long-duration expeditions require detailed examinations and evaluations of information flow dynamics, knowledge-sharing processes, and information technology use in distributed expert networks. This paper describes the work conducted with flight controllers in the Mission Control Center (MCC) of NASA's Johnson Space Center. This MCC work describes the behavior of experts in a distributed supervisory coordination framework, which extends supervisory control/command and control models of human task performance. Findings from this work are helping to develop analysis techniques, information architectures, and system simulation capabilities for knowledge sharing in an expert community. These findings are being applied to improve knowledge-sharing processes applied to a research program in advanced life support for long-duration space flight. Additional simulation work is being developed to create interoperating modules of information flow and novice/expert behavior patterns. PMID- 15835059 TI - The effect of the configuration and the interior design of a virtual weightless space station on human spatial orientation. AB - In a virtual weightless environment, subjects' orientation skills were studied to examine what kind of cognitive errors people make when they moved through the interior space of virtual space stations and what kind of visual information effectively decreases those errors. Subjects wearing a head-mounted display moved from one end to the other end in space station-like routes constructed of rectangular and cubical modules, and did Pointing and Modeling tasks. In Experiment 1, configurations of the routes were changed with such variables as the number of bends, the number of embedding planes, and the number of planes with respect to the body posture. The results indicated that spatial orientation ability was relevant to the variables and that orientational errors were explained by two causes. One of these was that the place, the direction, and the sequence of turns were incorrect. The other was that subjects did not recognize the rotation of the frame of reference, especially when they turned in pitch direction rather than in yaw. In Experiment 2, the effect of the interior design was examined by testing three design settings. Wall colors that showed the allocentric frame of reference and the different interior design of vertical and horizontal modules were effective; however, there was a limit to the effectiveness in complicated configurations. PMID- 15835060 TI - Hospital self-assessments indicate significant strides in medication safety efforts. PMID- 15835061 TI - Gravity and perceptual stability during translational head movement on earth and in microgravity. AB - We measured the amount of visual movement judged consistent with translational head movement under normal and microgravity conditions. Subjects wore a virtual reality helmet in which the ratio of the movement of the world to the movement of the head (visual gain) was variable. Using the method of adjustment under normal gravity 10 subjects adjusted the visual gain until the visual world appeared stable during head movements that were either parallel or orthogonal to gravity. Using the method of constant stimuli under normal gravity, seven subjects moved their heads and judged whether the virtual world appeared to move "with" or "against" their movement for several visual gains. One subject repeated the constant stimuli judgements in microgravity during parabolic flight. The accuracy of judgements appeared unaffected by the direction or absence of gravity. Only the variability appeared affected by the absence of gravity. These results are discussed in relation to discomfort during head movements in microgravity. PMID- 15835062 TI - California employers view health care cost-sharing as double-edged sword. PMID- 15835063 TI - Consent forms and the therapeutic misconception: the example of gene transfer research. PMID- 15835064 TI - Dual-frequency ultrasound for detecting and sizing bubbles. AB - ISS construction and Mars exploration require extensive extravehicular activity (EVA), exposing crewmembers to increased decompression sickness risk. Improved bubble detection technologies could help increase EVA efficiency and safety. Creare Inc. has developed a bubble detection and sizing instrument using dual frequency ultrasound. The device emits "pump" and "image" signals at two frequencies. The low-frequency pump signal causes an appropriately-sized bubble to resonate. When the image frequency hits a resonating bubble, mixing signals are returned at the sum and difference of the two frequencies. To test the feasibility of transcutaneous intravascular detection, intravascular bubbles in anesthetized swine were produced using agitated saline and decompression stress. Ultrasonic transducers on the chest provided the two frequencies. Mixing signals were detected transthoracically in the right atrium using both methods. A histogram of estimated bubble sizes could be constructed. Bubbles can be detected and sized transthoracically in the right atrium using dual-frequency ultrasound. PMID- 15835065 TI - The quality of informed consent in a clinical research study in Thailand. PMID- 15835066 TI - The promise and limits of international bioethics: lessons from the recent revision of the Declaration of Helsinki. PMID- 15835067 TI - Ethical guidelines for HIV research: a contextual implementation process. PMID- 15835068 TI - Draft additional protocol to the Convention on Human Rights and Biomedicine, on biomedical research. PMID- 15835069 TI - World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. PMID- 15835070 TI - Normativity and biomedicine in the USA. PMID- 15835071 TI - Towards an international bioethics law. PMID- 15835072 TI - International law and life sciences: a new battlefield of power or a new temple for justice and peace? PMID- 15835073 TI - Bioethics in Italy: two reasons of discontent. PMID- 15835074 TI - Informed consent in Italy: a Mediterranean country towards an autonomy-based model. PMID- 15835075 TI - Elaboration of the Declaration on Universal Norms on Bioethics: third outline of a text. PMID- 15835076 TI - Age-specific incidence and clearance of high-risk human papillomavirus infections in women in the former Soviet Union. AB - Recently, conflicting results on human papillomavirus (HPV) clearance have been reported and the data on the accumulation of incident HPV infections are still fragmentary. Thus, we completed an analysis of the age-specific incidence and clearance rates of high-risk (HR) HPV infections in 448 women participating in a multi-centre screening study in three New Independent States countries. At baseline, 239 of the 448 women were negative for HR HPV DNA, whereas 209 were HR HPV-positive and cleared HR HPV during the prospective follow-up. The cumulative incidence and clearance of HR HPV were modelled using life-table techniques. The monthly incidence rates of HR HPV were significantly age dependent (p = 0.0001), whereas monthly clearance rates remained constant across the nine age groups (p = 0.920). The incidence rates (3.04% and 2.65%) exceeded the clearance rates in the two youngest age groups only, 15-20- and 21-25-year-old women, and remained lower (0-0.84%) in all other age groups. The cumulative rate of incident HR HPV infections (1.0%) was significantly lower than the overall clearance rate (1.9%) (p = 0.001). In life-table analysis, incident HR HPV infections between the nine age groups were significantly different (p = 0.0001), while cumulative HR HPV clearance was identical in all groups (p = 0.822). The accumulation of incident HR HPV infections is significantly age-related, whereas virus clearance remains constant between 15 and 60 years of age. These distinct age-specific incidence and clearance rates explain the differences in age-specific prevalence of HR HPV infections in the study population. PMID- 15835077 TI - Immunology of intraocular tumors. AB - The immune surveillance hypothesis was introduced over 30 years ago and proposed that neoplasms express novel antigens that subjected them to immune detection and elimination. In order for immune surveillance to be effective in controlling neoplasms, two requirements must be satisfied: 1) the tumor must arise in a body site that permits the induction the full array of immune responses and 2) the immune elements generated must have unfettered access to the tumor and be able to express their entire range of effector functions at the tumor site. The unique immunologic and anatomic features of the eye prevent the induction and expression of conventional immunity--a phenomenon known as 'immune privilege'. Although ocular immune privilege represents a theoretical obstacle to immune surveillance, some highly immunogenic intraocular tumors can circumvent immune privilege and undergo immune rejection. Uveal melanoma is the most common intraocular malignancy in adults, yet it occurs with a frequency that is no higher than neoplasms arising in conventional bodies. The presence of either tumor infiltrating lymphocytes (TIL) or tumor-infiltrating macrophages (TIM) is associated with poor prognosis in uveal melanoma patients and suggests that some immune responses to intraocular tumors might exacerbate, rather than mitigate, tumor progression. Although counterintuitive, this proposition is consistent with the 'immune stimulation' hypothesis of tumor progression offered by Richmond Prehn over thirty years ago. It remains to be ascertained if immune stimulation affects the malignancy of ocular tumors, but it represents an intriguing explanation for the paradoxes of uveal melanoma. PMID- 15835079 TI - Politics, culture, and the lab: public attitudes toward animals and human embryonic stem cells have fostered different research agendas in Britain and the United States. PMID- 15835080 TI - NIH consultant finds little evidence of bias against clinical researchers. PMID- 15835078 TI - Hoping to avoid brain drain, states push to finance stem-cell research. PMID- 15835081 TI - Hyperammonemia: are the burdens too grave? Case study. PMID- 15835082 TI - "It's a bridge between science and people". Interview by David Crouch. PMID- 15835083 TI - Intellectual disability, genetics, and ethics: a review. PMID- 15835084 TI - Consent and persons with marginal mental impairment. PMID- 15835085 TI - Driving cardiac care. Interview by Janis Smy. PMID- 15835086 TI - The wisdom of Costa Rica. PMID- 15835087 TI - Pain definitions revised: newborns not only feel pain, they also suffer. AB - Premature babies are relatively unreactive, often being completely isolated in an incubator and considered incapable of social behavior. To the attentive observer, however, they reveal an unsuspected emotional world: not only do they feel pain (the clash arising from an attack to their physical integrity), but they are also capable of suffering (the clash arising from an attack to their person's integrity) as they are persons. In this paper the author argues in favour of newborns' personhood. This needs to be considered in neonatal analgesic treatment, even for extremely small babies, as recognition and adequate treatment of neonatal pain is still resisted: full acknowledgement of neonatal dignity and personhood is a prerequisite for an effective treatment of neonatal pain. PMID- 15835088 TI - Nurse-to-patient ratios: research and reality. PMID- 15835089 TI - Immediate animation: Thomistic principles applied to Norman Ford's objections. AB - Norman Ford and other scholars argue that the human embryo cannot have a human soul until after week two of fertilization. They claim that the individuality proper to ensoulment requires the development of the primitive streak and excludes the possibility of twinning. Their arguments are refuted by principles of philosophical realism inspired by Aristotle and Aquinas. According to these principles, the new substance formed by the union of the paternal and maternal DNA has a rational soul. The zygote, to be human, requires a human soul that determines its human development. The primitive streak is part of this development, not a condition of it. Twinning does not disprove the existence of a previous embryo with a human soul. Instead it indicates the existence of a new embryo derived from some totipotent cells that separate from the first embryo. PMID- 15835090 TI - Can artificial techniques supply morally neutral human embryos for research? Part I. Creating novel categories of human embryos. AB - Manipulations of the molecular composition and formation of human embryos are posing vital new challenges to traditional concepts of human identity and procreation. Current trends in embryology in particular are reshaping the ethical question of how scientific research should treat experimentally derived embryos. Some investigators have argued that embryos created through artificial means are technologically novel entities that should be exempt from ethical restraints placed on research involving human embryos that come into being through natural processes. These include uniparental embryos derived through cloning or parthenogenesis, as well as multiparental, hybrid-parental, and xenohybrid parental embryos. If confined to natural means many of these genetic unions could not occur, but through the intervention of technology, it is becoming possible to design and grow strange and unusual forms of embryos, in some cases using human gametes. Regardless of the genetic contributors or the processes used to fertilize and stimulate egg activation, in each case the new embryo represents an individual organism that begins a process of development. We conclude that the prospect of creating or redesigning new human life should be held to a stringent ethical standard of precaution, even higher than that of deciding to destroy existing embryonic life. Accordingly, we urge cautious ethical reflection and broad public discussion prior to deciding whether to permit embryologic research into novel forms of procreative means in nonhuman animals, to be further extended to humans. PMID- 15835091 TI - Lebensunwertes Leben: the devolution of personhood in the Weimar and pre-Weimar era. AB - Radical social change in most societies does not typically occur "overnight." It requires preparation--preparation in the way people think. The rise of euthanasia in western culture is a case in point. In order for assisted death to be increasingly accepted, the social-moral consensus must be altered. Typically, that consensus will be influenced by trends in biology, the behavioral sciences, ethics, law, even economics. Examining the manner in which assisted death was "prepared" in the decades before the ascendency of National Socialism is highly instructive as we witness the rise of euthanatic thinking in our own day several generations removed. PMID- 15835092 TI - 3TP and magnetic resonance breast imaging: interview with Dr. Julian Safir. PMID- 15835093 TI - How then should we do medical research? PMID- 15835094 TI - Integrative tumor board: transitional cell carcinoma of the bladder. PMID- 15835095 TI - In search of true value. PMID- 15835096 TI - Helping and impaired colleague. PMID- 15835097 TI - The past, present, and future of pediatric neurosurgery. Matson lecture, May 4, 2004. PMID- 15835098 TI - Assessing the validity of the endoscopic shunt insertion trial: did surgical experience affect the results? AB - OBJECT: Most surgical procedures are associated with a learning curve in which the success rate is lower early in the experience before mistakes have been identified and modifications made to the procedure. Negative results obtained early in a trial's learning curve may be a matter of timing rather than a reflection of the procedure's effectiveness. The recently published results of the Endoscopic Shunt Insertion Trial (ESIT) represent the notion that endoscopically placed shunts were no more likely to survive than conventionally placed shunts. This negative result may be due to inexperience in performing endoscopic surgeries. METHODS: . Surgical experience was assessed in two ways. Shunt survival rates were compared between cases treated endoscopically in the 1st and last years of the ESIT. The effect of center volume was evaluated using a Cox proportional hazard model in which the following variables were analyzed: age at registration, the diagnosis of myelomeningocele, head size, method of shunt placement (endoscopic compared with conventional), and center volume. There was no difference in survival (endurance) of the shunt between patients enrolled in the 1st and last years (log rank = 0.08, p = 0.77). Likewise, no variable in the Cox multivariate model, including center volume, was a significant factor in predicting shunt survival. CONCLUSIONS: The primary result of the ESIT was found to be internally valid. The fact that endoscopic shunt placement did not benefit patients evaluated in the study was not due to early timing of the trial. Any learning curve among the participating surgeons did not adversely affect the results. PMID- 15835099 TI - An instrument to measure the health status in children with hydrocephalus: the Hydrocephalus Outcome Questionnaire. AB - OBJECT: In the measurement of clinical outcome in pediatric patients with hydrocephalus the condition's effects on a child's physical, emotional, cognitive, and social health are frequently ignored. The authors developed a quantitative health status measure, the Hydrocephalus Outcome Questionnaire (HOQ), designed specifically for children with hydrocephalus, which can be completed by the children's parents. METHODS: The standardized steps in the development of a health status measure were followed. Item generation required involvement of health professionals and focus groups with parents of children with hydrocephalus. A comprehensive list of 165 unique health status items was thus generated. To streamline the list, questionnaires were sent to 69 sets of parents to solicit their opinions regarding the most important of these health issues, and the 51 most significant items were then selected to represent the following health domains: physical, social-emotional, and cognitive. In another cohort of 90 sets of parents, the 51-item questionnaire was then tested for reliability and construct validity against the following independent measures of specific components of health: Health Utilities Index, Wide Range Achievement Reading Test, Strengths and Difficulties Questionnaires, and Functional Independence Measure for Children. The HOQ took approximately 10 to 15 minutes for the parents to complete and demonstrated excellent test-retest reliability (0.93, 95% confidence interval [CI] 0.88-0.96), interrater reliability (0.88, 95% CI 0.79-0.93), and internal consistency (Cronbach alpha 0.94). Pearson correlation testing demonstrated very good construct validity between domain scores and their respective independent measures. CONCLUSIONS: The HOQ for children with hydrocephalus demonstrated excellent reliability and validity properties. This tool will be valuable for a wide range of clinical research projects in pediatric hydrocephalus. PMID- 15835100 TI - Measuring the health status of children with hydrocephalus by using a new outcome measure. AB - OBJECT: In the preceding article, the authors described the Hydrocephalus Outcome Questionnaire (HOQ), a simple, reliable, and valid measure of health status in children with hydrocephalus. In the present study, they present their initial experience in using the HOQ to quantify the health status in a typical cohort of children with hydrocephalus. METHODS: The mothers of children with hydrocephalus completed the HOQ and, with the child's attending surgeon, provided a global rating of their children's health. An exploratory analysis was performed using a multivariate analysis of variance (ANOVA) to determine which variables might be associated with worse health status. The mothers of 80 children, ranging in age from 5 to 17 years, participated in the study. The mean HOQ Overall Health score was 0.68, a value estimated to be equivalent to a mean health utility score of 0.77. The global health ratings provided by the mothers and the surgeons were moderately correlated with the HOQ scores (Pearson correlations 0.58 and 0.57, respectively). Results of the multivariate ANOVA indicated that the presence of epilepsy was strongly associated with a worse health status (p < 0.0001, F-test). CONCLUSIONS: The health status of a typical sample of children with hydrocephalus was measured using the HOQ. The only consistently significant association with health status found was the presence of epilepsy. PMID- 15835101 TI - The use of quick-brain magnetic resonance imaging in the evaluation of shunt treated hydrocephalus. AB - OBJECT: Children with shunt-treated hydrocephalus are exposed to serious amounts of radiation when undergoing computerized tomography (CT) scanning. The authors report their clinical experience with single-shot fast-spin echo (SSFSE) (quick brain) magnetic resonance (MR) imaging as the modality of choice for the workup and follow up of patients in whom a shunt has been placed to treat hydrocephalus. METHODS: A retrospective chart review was performed to obtain data on all cases in which a quick-brain MR image was acquired for either symptomatic workup or asymptomatic follow-up examination of shunt-treated hydrocephalus. Data regarding demographics, origin of hydrocephalus, MR imaging indications and findings, use of sedation, imaging-related complications, use of adjunctive CT scanning, details of shunt revision, and cause of shunt malfunction were collected. The authors found that SSFSE MR imaging is a sufficient, radiation-free diagnostic alternative to CT scanning that minimizes movement artifact and duration of scanning and eliminates the need for sedation. CONCLUSIONS: In light of these findings, the authors propose that quick-brain MR imaging replace CT scanning as the diagnostic modality of choice in examining and following shunt-treated patients because it offers significant advantages. PMID- 15835102 TI - Critical risk factors for intellectual impairment in children with posterior fossa tumors: the role of cerebellar damage. AB - OBJECT: Intellectual impairment is a major concern after treatment of malignant posterior fossa tumors in children. The effects of age at diagnosis and radiotherapy have been widely documented. Little is known, however, about perioperative factors, especially neurological damage to the cerebellum, the role of which in cognition and learning has been recently indicated. The authors studied the effects in 76 children treated for a malignant posterior fossa tumor in a cross-sectional study. METHODS: Two thirds of the tumors were medulloblastoma. Neuropsychological evaluation was performed at least 6 months after the end of treatment, and findings were correlated with clinical risk factors for intellectual impairment. The mean verbal intelligence quotient (VIQ) score was 87 +/- 19 (+/- standard deviation) and the mean performance IQ (PIQ) score was 76 +/- 17.5. A single neuropsychological test measuring hand skills (the Purdue Pegboard) was the strongest predictor of low IQ scores including items testing higher cognitive functions. A low VIQ was associated with impaired hand skills (p < 0.0001) and the presence of preoperative hydrocephalus (p = 0.02), whereas a low PIQ was associated with impaired hand skills (p < 0.0001) and incision of the vermis (p = 0.02). Impaired hand skills were associated with postoperative cerebellar mutism, oculomotor deficits, cerebellar syndrome, and therapeutic requirements. CONCLUSIONS: When treatment schedules are adapted to risk of disease and age, surgery-related risk factors then become critical for predicting intellectual impairment. Children with cerebellar damage are particularly at risk for long-term neuropsychological dysfunction and require active rehabilitation measures. Reducing surgery-related morbidity should be the next goal to reduce posterior fossa surgery-specific deficits. PMID- 15835104 TI - Foramen magnum cerebrospinal fluid flow characteristics in children with Chiari I malformation before and after craniocervical decompression. AB - OBJECT: The Chiari I malformation presents significant challenges to clinicians because its pathophysiology is not well understood. In conducting cerebrospinal fluid (CSF) flow studies, investigators have attempted to correlate the clinical severity of these lesions with general flow velocity or bulk flow at the foramen magnum; however, these techniques have not allowed consistent prediction of symptomatology, explanation of the presence of syringomyelia, or the assessment of the hydrodynamic characteristics of the decompression. The authors used temporally and spatially resolved flow analyses to assess the characteristics of CSF flow in children with Chiari I malformation and the changes in these flow characteristics that occur after suboccipital decompression. METHODS: The authors studied eight children with symptomatic Chiari I malformation with or without syringomyelia and two children without Chiari I malformation. All patients underwent phase-contrast magnetic resonance imaging before and after posterior fossa decompression. Velocity plots were displayed for each voxel. Several indices of CSF flow were developed to characterize the flow patterns associated with Chiari I malformation. In children with symptomatic Chiari I malformation, even though bulk flow or velocity is often normal, there was marked heterogeneity of flow at the foramen magnum. This was evident for several reasons: 1) an increase in cephalad and caudad peak velocities; 2) spatial inhomogeneity in velocities; 3) simultaneous bidirectional flow; and 4) substantial net craniad or caudad flows within particular voxels and subregions during the cardiac cycle. After posterior fossa decompression, the severity of these flow abnormalities decreased. CONCLUSIONS: Foramen magnum CSF flow in children with symptomatic Chiari I malformations is spatially and temporally heterogeneous, and this heterogeneity improves postoperatively. The authors propose that relying on mean flow parameters in patients with Chiari I malformation is no longer sufficient; instead, more elaborate techniques to analyze foramen magnum CSF flow have become necessary. PMID- 15835103 TI - The effects of hydrocephalus on intelligence quotient in children with localized infratentorial ependymoma before and after focal radiation therapy. AB - OBJECT: The goal of this study was to determine the influence of hydrocephalus on intelligence quotient (IQ) in children with infratentorial ependymoma before and after the administration of focal radiation. METHODS: Measurements of ventricular size, including Evans index (EI), cella media index (CMI), frontal horn diameter (FHD), and ventricular angle, were performed using magnetic resonance imaging at the time of diagnosis and again at 3, 6, 9, and 12 months after the initiation of radiation therapy. Of the 59 patients (median age at time of radiation treatment, 4.1 years), the clinical diagnosis established in 50 (85%) was hydrocephalus and 23 (39%) required placement of a cerebrospinal fluid (CSF) shunt. Extent of resection was gross or near total in 50 (85%). Before and after radiation treatment, IQ was measured using age-appropriate testing. The correlation between multiple ventricular measurements and IQ was investigated using standard regression techniques and a generalized linear model. Patients with a higher EI (p = 0.04), CMI (p = 0.001), and FHD (p = 0.0002) at the time of diagnosis were more likely to have lower IQ scores before radiation treatment. Patients with higher CMI (p = 0.04) and FHD (p = 0.01) at the time of diagnosis were more likely to exhibit an increase in IQ score after radiotherapy. The rate of change in IQ after radiation treatment was positively correlated with the CMI intercept (p = 0.015) and negatively correlated with the rate of FHD change (p = 0.006). CONCLUSIONS: Changes in IQ score before and after radiation treatment are significantly influenced by the extent and treatment of hydrocephalus at the time of diagnosis. Hydrocephalus is an important factor to include when analyzing the effects of treatment. Patients who undergo a second surgery for ependymoma are more likely to require the placement of a CSF shunt (p = 0.0001). PMID- 15835105 TI - A critical analysis of the Chiari 1.5 malformation. AB - OBJECT: Although the traditional nomenclature used to describe hindbrain hernias is useful, there are certainly patients in whom these morphological entities do not strictly apply. One such group is composed of patients with the more recently described Chiari 1.5 malformation in which a Chiari I malformation is seen in combination with brain-stem herniation through the foramen magnum. In an attempt to elucidate further the best surgical strategy and to refine the descriptive terminology, the authors retrospectively analyzed all cases at their institutions in which this form of hindbrain herniation was diagnosed. METHODS: The authors reviewed the database for all patients in whom Chiari 1.5 malformation had been diagnosed. Each patient had undergone a posterior fossa decompressive surgery. Magnetic resonance images were evaluated for the extent of caudal descent of the brainstem, amount of tonsillar ectopia, inclination of the odontoid process, and any brain or brainstem abnormalities. Clinical presentations and postoperative results were correlated to the aforementioned radiological findings. Twenty-two patients were identified. The obex was a mean 14.4 mm inferior to the foramen magnum, and the medulla exhibited a flattened appearance in the midsagittal plane in all patients. Syringomyelia was documented in 50% of the cases. The cerebellar tonsils were found to lie at C-1 and C-2 in nine and 13 patients, respectively. The mean angulation of the odontoid process was 84.4 degrees. No abnormalities or caudal descent of the midbrain or pons was identified. Eighteen patients experienced resolution of preoperative symptoms. Persistence of syringomyelia prompted a second posterior fossa operation secondary to progressive scoliosis in 13.6% of the patients. CONCLUSIONS: No single sign or symptom was found to be peculiar to the Chiari 1.5 malformation, although all patients in whom this diagnosis was established had undergone a posterior fossa decompressive surgery. A significant number (13.6%) of patients required repeated operation for persistent syringomyelia. Neurosurgeons may wish to consider that many patients may not respond as well to posterior fossa decompressive surgery especially if syringomyelia is present. PMID- 15835106 TI - Dura-splitting decompression of the craniocervical junction: reduced operative time, hospital stay, and cost with equivalent early outcome. AB - OBJECT: The choice of surgical technique for decompressive surgery in patients with Chiari I malformation is controversial. Good preliminary postoperative outcomes have been achieved in patients with Chiari I malformation (without syringomyelia) after using a dura-splitting technique. The authors evaluated safety, resource use, and early outcome after this surgery in patients without syringomyelia and compared the findings associated with duraplasty in patients with syringomyelia. METHODS: A prospective series of 24 patients with Chiari I malformation (12 with a syrinx) underwent decompression of the craniocervical junction (CCJ). An allograft-augmented duraplasty was performed in patients with syringomyelia. Intraoperative ultrasonography confirmed adequate tonsillar decompression after lysis of the periosteal bands at the foramen magnum and C-1 arch as well as partial resection of the outer leaf of the dura in patients without syringomyelia. Patients in each group were of similar mean age (syringomyelia 10.8 years and no syringomyelia 7.6 years old; p = 0.07) and functional status. The mean follow-up period was 15.3 months (range 3-30 months). Dura-splitting decompression required significantly less mean operative time (99 minutes compared with 169 minutes, respectively; p < 0.001), total operating room time (166 minutes compared with 249 minutes, respectively; p < 0.001), duration of hospitalization (3 days compared with 3.75 days, respectively; p < 0.05), perioperative charges ($3615 compared with $5538, respectively; p < 0.001), and overall hospital charges ($7705 compared with $9759, respectively; p < 0.001) than the duraplasty. Mean clinical outcome scores were similar (syringomyelia 1.53 of 2; no syringomyelia 1.67 of 2; not statistically significant). CONCLUSIONS: Dura-splitting CCJ decompression in pediatric patients with Chiari I malformation and without syringomyelia is safe, provides good early clinical results, and significantly reduces resource use. A randomized controlled trial of dura-splitting decompression in a uniform population of patients with Chiari I malformation is indicated. PMID- 15835107 TI - Treatment of basilar invagination associated with Chiari I malformations in the pediatric population: cervical reduction and posterior occipitocervical fusion. AB - OBJECT: The reduction of basilar invagination associated with Chiari I malformations in pediatric patients is often possible. Eleven children ranging in age from 1.5 to 17 years underwent a novel treatment method involving decompression, manual reduction, and posterior instrumentation-augmented fusion. Outcomes were evaluated retrospectively. METHODS: After decompression of the lesion, manual craniocervical distraction and extension were performed, followed by posterior occipitocervical fusion involving rigid internal fixation. All patients were symptomatic at presentation. Intraoperative monitoring included somatosensory evoked potentials, fluoroscopy, and direct intradural demonstration of the craniocervical junction. No new neurological deficits occurred immediately after surgery. Perioperatively, symptoms in seven patients improved significantly and in four they remained unchanged. Three patients required subsequent transoral resections. During long-term follow up (mean 39.4 months, range 3-92 months) symptoms improved markedly in nine, remained unchanged in one, and progressively worsened in one patient. CONCLUSIONS: In selected cases, manual cervical distraction and extension, posterior fixation, and fusion appear to provide a safe, effective standalone treatment for basilar invagination associated with Chiari malformation in children. PMID- 15835108 TI - Effects of hydrocephalus and ventriculoperitoneal shunt therapy on afferent and efferent connections in the feline sensorimotor cortex. AB - OBJECT: The authors of previous studies have suggested that connectivity within the cerebral cortex may be irreversibly altered by hydrocephalus. To examine connectivity-related changes directly, the authors conducted a study in which they used an axonal tracer in an animal model of infantile hydrocephalus. METHODS: In five hydrocephalic kittens low-pressure ventriculoperitoneal (VP) shunts were placed 10 to 14 days after induction of hydrocephalus by intracisternal kaolin injections. Wheat germ agglutinin-conjugated horseradish peroxidase was injected laterally into the motor cortex in hydrocephalic animals 9 to 15 days after kaolin injection, and 1, 2, and 4 weeks after VP shunt insertion in shunt-treated animals, and in age-matched controls. Reduction of antero- and retrograde labeling was most profound within the contralateral cortex and portions of the midbrain. Thalamic nuclei exhibited reductions in anterograde and retrograde labeling. Labeling within cell bodies of the ventral tegmental area decreased greatly in animals with untreated hydrocephalus, in which retrograde labeling was reduced in the locus coeruleus but did not affect the raphe nucleus. Shunt treatment increased both antero- and retrograde labeling of contralateral motor cortex to near-normal levels. Thalamic relay nuclei recovered antero- and retrograde labeling, although not to levels exhibited in controls. Shunt therapy restored cellular labeling within the ventral tegmental area and locus coeruleus. Recovery of labeling occurred as early as 7 days after shunt insertion. CONCLUSIONS: Collectively, analysis of these data indicates the following. 1) Cortical connectivity involving both afferent and efferent pathways was impaired in untreated hydrocephalic animals. 2) Shunt therapy improved both cortical afferent and efferent connectivity. 3) Complete reestablishment of the cortical efferent pathways, however, did not occur. Cortical pathway dysfunction, if permanent, could cause many of the motor and cognitive deficits seen clinically in children with hydrocephalus. PMID- 15835109 TI - Acquired Chiari I malformation following baclofen pump placement in a child. Case report. AB - The authors present a case of a child suffering from shunt-treated hydrocephalus and spastic quadriplegia who underwent surgery for placement of a baclofen pump. Magnetic resonance (MR) imaging performed prior to pump placement demonstrated no hindbrain herniation. Afterward, however, the patient exhibited symptoms of brainstem compression, and MR imaging revealed a significant Chiari I malformation along with a fully functioning ventriculoperitoneal shunt. Posterior fossa decompression was performed, and the patient's symptoms abated. The authors believe this to be the first report of an acquired Chiari I malformation in a patient with a baclofen pump. Clinicians should consider Chiari I malformation as a rare but severe complication of baclofen pump placement. PMID- 15835110 TI - Subtemporal decompression for slit-ventricle syndrome: successful outcome after dramatic change in intracranial pressure wave morphology. Report of two cases. AB - Subtemporal decompression is recognized as an effective treatment for slit ventricle syndrome; however, the effects of this procedure have not been demonstrated using both pre- and postsurgical intracranial pressure (ICP) monitoring. The authors report two cases in which slit-ventricle syndrome and elevated ICP had been diagnosed. Each patient underwent ICP monitoring before and after subtemporal decompression; the dramatic changes in the ICP measurements are presented along with findings from 1-year follow-up examinations. PMID- 15835111 TI - Frontoethmoidal encephalocele with ocular leakage of cerebrospinal fluid. Case report. AB - The authors present the case of a 2-year-old boy who was born with a small bulging mass in the middle frontonasal area and hypertelorism. In the neonatal period he suffered from a continuous lacrimal secretion mistaken for recurrent conjunctivitis. Cranial computerized tomography scanning and magnetic resonance imaging revealed a frontoethmoidal encephalocele associated with an ocular leakage of cerebrospinal fluid. One-stage repair of the encephalocele, along with correction of hypertelorism and bone grafting of the forehead, was performed with good result. During a 2-year follow-up period no neurological deficits appeared. PMID- 15835112 TI - Adjustment and malfunction of a programmable valve after exposure to toy magnets. Case report. AB - Inadvertent adjustments and malfunctions of programmable valves have been reported in cases in which patients have encountered powerful electromagnetic fields such as those involved in magnetic resonance imaging, but the effects of small magnetic fields are not well known. The authors present a case in which a child playing with a collection of commercially available toy magnets altered the pressure setting of an implanted valve and may have caused its permanent malfunction. PMID- 15835113 TI - Spontaneous regression of congenital spinal lipomas of the conus medullaris. Report of two cases. AB - Spinal lipomas of the conus (SLCs) are relatively common closed neural tube defects. Many of these lesions are treated surgically at the time of diagnosis and, therefore, little has been documented about their true natural progression. In addition, the changes, if any, of SLCs in pediatric patients have not been widely studied or documented using magnetic resonance (MR) imaging. In this report, the authors describe two asymptomatic infants in whom serial MR imaging revealed substantial spontaneous regression of SLCs, a phenomenon that has been very rarely reported in the literature. These are the first MR imaging-documented cases of near-complete spontaneous regression of SLCs in infants. PMID- 15835114 TI - The relationship of the spinal cord to scoliosis. AB - The authors present an original description of the tethered spinal cord. This submission in its day was not welcomed by the contemporaries of Dr. Robert Heimburger who theorized that there may be an association between the tethered spinal cord and scoliosis. Today we know that abnormal curvatures of the spinal cord can result from distal abnormal tensions placed on the spinal cord. This historical paper, now printed in full, may very well be one of the earliest descriptions of this association. It offers a unique window into the early cause and-effect thinking regarding this neurosurgical lesion. PMID- 15835115 TI - Surface charge influence on the surface plasmon absorbance of electroactive thiol protected gold nanoparticles. AB - Gold nanoparticles (3.1-5.0 nm in size) surface-derivatized with both electroactive and nonelectroactive self-assembled monolayers were synthesized. The surface-derivatized electroactive particles can be easily oxidized/reduced at an electrode surface based on the diffusion-controlled current-voltage curve observed in cyclic voltammetry measurements. Spectroelectrochemical investigation demonstrated that the maximum absorbance of the nanoparticles in their oxidized state red-shifted compared with their reduced state to a different extent according to their size distribution. In the case of the particles surface derivatized with nonelectroactive monolayers, much less shift was observed. This study showed that surface plasmon absorbance of gold nanoparticles was not only related to core charge states but was also influenced by surface charge states as well. PMID- 15835116 TI - Ultrahigh surface area nanoporous silica particles via an aero-sol-gel process. AB - We describe a new salt-assisted aero-sol-gel approach to produce spherical nanosized mesoporous silica particles. As an alternative to expensive templating mediums in prior works, salt (NaCl) was employed as a templating medium because it is thermally stable, recyclable, and easily leached. Furthermore, we demonstrate the ability to carry out traditional sol-gel chemistry within an aerosol droplet. The role of salt in sol-gel chemistry and aerosol processing was investigated as a function of hydrolysis time. It was verified that salt accelerates the kinetics of silica gelation, and simultaneously becomes an excellent templating medium to support nano-sized pores inside silica structures in the aerosol processing route. The presence of salt results in a roughly ten fold increasing in the pore specific surface area and pore volume, subsequent to leaching of the salt matrix. The surface area and pore volume of the as-produced nanoporous silica particles was found to increase with increasing sol-gel hydrolysis time. PMID- 15835117 TI - Unique rodlike surface morphologies in trisilanolcyclohexyl polyhedral oligomeric silsesquioxane films. AB - A trisilanol derivative of polyhedral oligomeric silsesquioxane (POSS), trisilanolisobutyl-POSS, has recently been reported to form stable monolayers at the air/water interface. This paper explores the mono- and multilayer properties of another POSS derivative, trisilanolcyclohexyl-POSS, with pi-A isotherm and Brewster angle microscopy measurements. Results show that with continuously increasing surface concentration via symmetrical compression, trisilanolcyclohexyl-POSS amphiphiles at the air/water interface undergo a series of phase transitions from traditional Langmuir monolayers (one-POSS-molecule thick) to unique rodlike hydrophobic aggregates in multilayer films (approximately eight-POSS-molecules thick) that are dramatically different from "collapsed" morphologies seen in other systems. Stable and hydrophobic rodlike structure formation on water is presumably due to trisilanolcyclohexyl-POSS' unique molecular structure and strong tendency to form intermolecular hydrogen bonds in the solid state. This result is consistent with existing POSS/polymer composite research, which shows that POSS molecules tend to aggregate and crystallize into lamellar nanocrystals. PMID- 15835118 TI - Pattern generation of biological ligands on a biodegradable poly(glycolic acid) film. AB - The micropatterns of biological ligands (biotin and RGD peptides) were generated on a flat surface of biodegradable polymer, poly(glycolic acid) (PGA). The immobilization of biological ligands onto the surface of biodegradable polymers (especially aliphatic polyesters) is usually hampered by the absence of functionalizable groups on the polymer backbone. We demonstrate herein that PGA polymer films were modified by surface hydrolysis to introduce carboxylic acid groups on the film surfaces, which were subsequently used for patterning amine terminated ligands by microcontact printing. Fluorescence microscopy was used to verify the pattern of biotin on the surface of the PGA films after complexation with fluorescein-conjugated streptavidin. In addition, the cellular micropatterns were obtained from micropatterns of RGD peptides on the surface-hydrolyzed PGA films. PMID- 15835119 TI - Antioxidant action by gold-PAMAM dendrimer nanocomposites. AB - Gold-dendrimer nanocomposites were prepared in the presence of poly(amidoamine) (PAMAM) dendrimer (generation 3, 3.5, 5, and 5.5) via reduction of HAuCl4 with sodium borohydride. The average particle size of the gold nanoparticles was independent of the dendrimer concentration, ranging between 3.0 and 4.3 nm in diameter. The catalytic activities of the gold-dendrimer nanocomposites upon elimination of hydroxyl radicals formed in an H2O2/FeSO4 system were examined using a spin-trapping method. The gold-dendrimer nanocomposites exhibited high catalytic activities which were hardly affected by the concentration and the generation of the dendrimer except PAMAM dendrimer 3.5. The highest activity for the gold-PAMAM dendrimer 3.5 nanocomposites was 85 times that of ascorbic acid. PMID- 15835120 TI - Complexation of polyacrylates by Ca2+ ions. Time-resolved studies using attenuated total reflectance Fourier transform infrared dialysis spectroscopy. AB - The attenuated total reflectance Fourier transform infrared dialysis technique is introduced for the time-resolved investigation of the binding processes of Ca2+ to polyacrylates dissolved in water. We observed transient formation of intermediates in water with various types of coordination of the carboxylate group to Ca2+ throughout the complexation steps. Time-resolved changes in the spectra were analyzed with principal component analysis, from which the spectral species were obtained as well as their formation kinetics. We propose a model for the mechanisms of Ca2+ coordination to polyacrylates. The polymer chain length plays an important role in Ca2+ binding. PMID- 15835121 TI - Kinetics of protein aggregation with formation of unreactive intermediates. AB - Irreversible protein aggregation resulting in formation and deposition of insoluble fibrils or amorphous precipitates is usually assumed to occur via sequential attachment of monomers to soluble intermediates. We complement this scheme by slow conversion of the intermediates to a relatively stable form so that they do not react with monomers but can be trapped by precipitates. For reasonable values of parameters, our model predicts that the aggregation kinetics order may be between 2.0 and 2.5. In particular, the model can be used to explain the reaction order, 2.17 +/- 0.09, observed for aggregation of recombinant human granulocyte colony stimulating factor. PMID- 15835122 TI - Swelling-shrinking behavior of a polyampholyte gel composed of positively charged networks with immobilized polyanions. AB - Polyampholyte gels were prepared by free radical polymerization of aqueous monomer solutions with the following composition: 69% N-isopropylacrylamide (thermosensitive neutral monomer), 1% N,N'-methylenebisacrylamide (cross-linker), 15% 1-vinylimidazole (cationic monomer), and either 15% acrylic acid (AAc, anionic monomer) or poly(acrylic acid) (PAAc, polyanion). We thus obtained two sorts of polyampholyte gels; that is, G1 with immobilized PAAc and G2 with randomly copolymerized AAc. The equilibrium swelling ratio (Qe) was studied as a function of the pH, NaCl concentration, and temperature. Also studied was the kinetics of swelling and shrinking in response to a sudden pH change. The significant results obtained were as follows: (i) A fully collapsed state was observed at pH 4.5-9.0 for G1 and at pH 4.5-7.0 for G2. (ii) Below and above these pH ranges, both gels were in a swollen state; therefore, an isoelectric point (pI) appeared in a wide pH range. (iii) At alkaline pH regions where a hysteresis was observed in the Qe versus pH curves of G1 and G2 as the pH was first increased then decreased, G1 exhibited very slow swelling-shrinking kinetics. (iv) An increase in the NaCl concentration allowed the gel to swell at pH approximately pI (antipolyelectrolyte behavior) but to shrink at pHs below and above the pI range (polyelectrolyte behavior). (v) The magnitude of the salt induced shrinking of G1 is smaller than that of G2 at pH 10 and at NaCl concentrations > 0.01 M. (vi) At pH 10, an increase in the temperature from 35 to 50 degrees C led to a shrinking change of G1 but not of G2. These results were found to be explicable in terms of a different distribution of negative charges within the polyampholyte gel network. PMID- 15835123 TI - Programmable assembly of colloidal particles using magnetic microwell templates. AB - A substrate of thin micromagnets covered by a template of microwells is used to direct the assembly of superparamagnetic colloidal beads into two-dimensional arrays. It is confirmed that the magnetization of the micromagnets can direct beads to programmed locations on the substrate with assistance of externally applied magnetic fields. Empirical investigations on this topic were guided by mathematical models with the intent to elucidate the conditions that promote a single bead to be assembled in the desired microwells. To demonstrate that this technique is programmable, heterogeneous arrays of colored beads are produced. PMID- 15835124 TI - Interfacial properties of branch-tailed fluorinated surfactants yielding a water/supercritical CO2 microemulsion. AB - We have examined the interfacial properties of several fluorinated surfactants in a water/CO2 mixture with a pendant drop tensiometer and revealed the relationships between the interfacial properties, the surfactant structure, and the microemulsifying power. We employed the following Aerosol-OT analogue surfactants that have two fluorinated tails: bis(1H,1H,5H-octafluoropentyl)-2 sulfosuccinate (di-HCF4), sodium bis(1H,1H,9H-hexadecafluorononyl)-2 sulfosuccinate (di-HCF8), sodium bis(1H,1H,2H,2H-heptadecafluorodecyl)-2 sulfosuccinate (8FS(EO)2), and sodium bis((1H,1H,2H,2H-heptadecafluorodecyl) oxyethylene)-2-sulfosuccinate (8FS(EO)4). To discuss the effect of the fluorocarbon/hydrocarbon ratio in single surfactant molecules, water/CO2 interfacial tension (IFT) of a hybrid surfactant with one fluorocarbon and one hydrocarbon tail, that of a surfactant with a single fluorinated tail, and that of a hydrocarbon surfactant, Aerosol-OT (AOT), were examined. The hybrid surfactant employed was sodium 1-oxo-1-[4-(tridecafluorohexyl)phenyl]-2 hexanesulfonate (FC6-HC4), and the single-tailed surfactant was perfluoropolyether ammonium carboxylate (PFPECOONH4, CF3CF2(CF2OCF(CF3))4COONH4). All of the fluorinated AOT analogue surfactants exhibited an excellent level of activity at the water/CO2 interface compared with other fluorinated surfactants and AOT. With a larger hydrocarbon chain number in the CO2-philic tails (i.e., from 0 to 2), the IFT of the AOT analogue surfactants was increased. The area occupied by one surfactant molecule at the water/CO2 interface, A, and the critical microemulsion concentration, cmicroc, were determined and used to examine the water-to-surfactant molar ratio within a reversed micelle, W0c, of the surfactants. The surfactants that form W/scCO2 microemulsions with a large W0c were found to lower the interfacial tension efficiently irrespective of increases in temperature. To achieve the most desirable W0C, the surfactant needs not only a high CO2-philicity of the tails but also a high Krafft point, properties which induce a low hydrophilic/CO2-philic balance. PMID- 15835125 TI - Test of the Epstein-Plesset model for gas microparticle dissolution in aqueous media: effect of surface tension and gas undersaturation in solution. AB - The gas from a free air bubble will readily dissolve in water, driven by two main factors: the concentration (undersaturation) of dissolved gas in the aqueous solution and the surface tension of the gas bubble-water interface via a Laplace overpressure in the bubble that this creates. This paper experimentally and theoretically investigates each of these effects individually. To study the effects of surface tension, single- and double-chain surfactants were utilized to control and define interfacial conditions of the microbubble in saturated solution. To study the effect of undersaturation, solid distearoylphosphocholine lipid was utilized to coat the gas microparticle with, essentially, a wax monolayer and to achieve zero tension in the surface. The experimental work was performed using a micromanipulation technique that allows one to create and micromanipulate single air microparticles (5-50 microm radius range) in infinite dilution and to accurately record the size of the particle as it loses volume due to the dissolution process. The micropipet technique has shown to be an improvement over other previous attempts to measure dissolution time with a 3.2% average experimental error in gas microparticle dissolution time. An ability to study a gas microparticle in infinite dilution in an isotropic diffusion field is in line with the theoretical assumptions and conditions of the Epstein-Plesset model. The Epstein-Plesset model on average underpredicted the experimentally determined dissolution time by 8.6%, where the effect of surface tension was considered with a range of surface tensions from 72 down to 25 mN/m. The Epstein Plesset model on average overpredicted the dissolution time by 8.2%, where the effect of undersaturation was considered for a microparticle with zero tension in the surface (zero Laplace pressure) and a range of gas saturations from 70% to 100%. Compared to previous attempts in the literature, this paper more appropriately and accurately tests the Epstein-Plesset model for the dissolution of a single microbubble and an air-filled microparticle in aqueous solution. PMID- 15835126 TI - Nuclear magnetic resonance investigation of the micellar properties of two-headed surfactant systems: the disodium 4-alkyl-3-sulfonatosuccinates. 2. The dynamics of the chains comprising the interior of two-headed surfactant micelles. AB - The dynamics of the carbons comprising the micelles of two members of the family of two-headed surfactants, the disodium 4-alkyl-3-sulfonatosuccinates, has been determined via the application of the two-step model to the 13C relaxation rates and the nuclear Overhauser enhancements (nOe's) at 200 MHz. The NMR relaxation times, determined from the inversion recovery method, increase steadily as we descend the chain from the headgroup region. The relaxation rate profiles and the order parameters have been calculated from the two-step model for the micellar sulfosuccinate aggregates. We note that the order parameter profile and the fast motion correlation time profile for these two-headed surfactants are distinctly different from those of a typical single-headed, single-tailed surfactant such as dodecyltrimethylammonium bromide, particularly in the headgroup region of the micelle. All these results are interpreted in terms of the effect of adding a second headgroup to a single-headed, single-tailed surfactant. PMID- 15835127 TI - Quantitative effect of nonionic surfactant partitioning on the hydrophile lipophile balance temperature. AB - Phase behaviors of water/nonionic surfactants/isooctane systems are determined experimentally in temperature-global surfactant concentration diagrams. The surfactants are monodistributed polyoxyethylene glycol n-dodecyl ether. They are used as model mixtures of two, three, or five compounds or as constituents of a commercial surfactant. It is found that the phase diagrams of these systems are bent gradually toward the highest temperatures as the global surfactant concentration decreases. Each phase diagram is well-characterized by the curve of the HLB (hydrophile-lipophile balance) temperature versus the global surfactant concentration. For any fixed global surfactant concentration, this temperature is the middle temperature of the three-phase region; it can be calculated from an additive rule of the HLB temperatures of the surfactants weighted by their mole fractions at the water/oil interface. These mole fractions are determined through the pseudophase model using surfactant partitioning. Calculations require the knowledge of the critical micelle concentration, the partition coefficient between water and oil, and the HLB temperature of each surfactant of the mixture. This treatment can be used to correctly predict the variation of the HLB temperatures of the surfactant mixtures studied versus the global surfactant concentration. Furthermore, these calculations show that the observed curvature of the phase diagrams at the lowest global concentrations is due to the most favorable partitioning toward the oil of the lowest ethoxylated surfactant molecules. PMID- 15835128 TI - Phase behavior of CO2-expanded fluorinated microemulsions. AB - The formation of CO2-expanded, fluorinated reverse microemulsions is demonstrated for the system of perfluoropolyether (PFPE) surfactant (ClPFPE-NH4, MW = 632) and PFPE oil (PFPE, MW = 580). The phase behavior of this system is examined as a function of temperature (25-45 degrees C), pressure, CO2 concentration, and water to surfactant molar ratios (W0 = 10 and 20). Visual observations of one-phase behavior consistent with reverse microemulsion formation are further supported by spectroscopic measurements that establish the existence of a bulk water environment within the aqueous core. Microemulsion formation is not observed in the absence of CO2 for this PFPE surfactant/PFPE oil system, and a CO2 content greater than 70 mol % is required to induce microemulsion formation. Over the range of water loadings and temperatures investigated, the lowest cloud point pressure is observed at 46 bar (5 wt % ClPFPE-NH4 in PFPE oil, W0 = 20, xCO2 = 0.7, T = 25 degrees C). In the regions where one-phase behavior is observed, the cloud point pressures increase with temperature, water loadings, and CO2 content. The driving forces of microemulsion formation in the CO2-expanded fluorinated solvent are discussed relative to traditional reverse microemulsions and CO2 continuous microemulsions. PMID- 15835129 TI - Protecting nanoscaled non-oxidic particles from oxygen uptake by coating with nitrogen-containing surfactants. AB - To suppress the reactivity of nanoscaled non-oxidic powders of titanium nitride (TiN) and silicon carbonitride (SiCN) against hydrolysis and oxidation, chemical surface modification with nitrogen-containing surfactants was investigated. Among these surfactants, long-chain primary amines, ethylenediamines, guanidines, nitriles, isocyanates, and succinimides were examined. Thermogravimetry, elemental analysis, and behavior against the water-vapor adsorption of the modified particles were used as methods to estimate the protective capacity of the organic coating material. The best results were obtained by using the long chain amines and octadecylisocyanate, which were indicated by a significant shift of the powder oxidation toward the higher temperatures and an increase of the particle hydrophobicity. A long-chain succinimide was found to be the most effective in dispersing nanoscaled TiN in organic media. Preparation of a stable aqueous dispersion without significant changes in the elemental composition of the powder was achieved by the application of an ionic surfactant to the surface modified particles. PMID- 15835130 TI - Cubic phase prepared in an anionic/amphoteric surfactant/oleic acid/decane/water system and the relationship with the neighboring phase. AB - The formation, properties, and structure of discontinuous cubic phase in the pseudo-ternary system consisting of N'-carboxyethyl N'-hydroxyethyl N-aminoethyl dodecylamide (imidazoriniumbetain), sodium and triethanol amine salt of polyoxyethylene (1.5 mol) myristyl ether sulfate, oleic acid, decane, and water at a constant surfactant/water ratio of 4/6 were studied by means of small-angle X-ray scattering, freeze-fracture transmission electron microscopy, static light scattering, and dynamic rheology to gain an insight in its origin and interrelation with neighboring phases. It was found that the cubic phase occupied a rather wide region in a constructed ternary phase diagram, including from 25 to 45% of decane. Its properties and structural parameters varied with changing the oil content. The decane addition caused the swelling of spherical micellar aggregates. This resulted in an increase of their diameter up to 35 nm, which was ca. nine times larger than that of the initial micelles, and micellar volume fraction (packing fraction) up to 72 vol. %, which was close to the theoretically possible value of 74 vol. % for the close-packed spherical particles. The cubic phase was surrounded by a micellar L1 phase from the water-rich side (separated by a short two-phase region), two-phase region (cubic + oil) from the oil-rich side, and a lamellar phase from the surfactant-rich side. A transition from the L1 phase to the cubic state at the packing fraction of 60 vol. % was caused by an increase in the packing density of micellar aggregates, occurring with the decane addition. When it reached 72 vol. %, the oil started forming a separated phase owing to the inability of micelles to dissolve it. The important observation is that the adjacent phase from the surfactant-rich side was a lamellar one made up of flat bilayers. The preliminary data showed that the lamellar phase coexisted with cylindrical micelles in the intermediate two-phase region separating the cubic and lamellar phases. PMID- 15835131 TI - Electrophoresis of concentrically and eccentrically positioned cylindrical particles in a long tube. AB - We study analytically and numerically the electrophoretic motion of cylindrical particles translating slowly in long tubes filled with an electrolyte solution and subjected to axial electric fields. Both thin and thick double layers are considered. Of particular interest is the case when the particle's and tube's radii are of the same order of magnitude. The model accounts for the flow induced by the particle's motion (the particle acts as a leaky piston) and the electroosmotic flow in the tube. The electrophoretic velocity of the particle and the forces and torques acting on it are determined as functions of the particle's radius, length, and position, the particle's and tube's zeta potentials, the tube's length, and the externally imposed pressures. When the particle is positioned off center, it rotates and its trajectory traces an oscillatory path. PMID- 15835132 TI - Dendritic chelating agents. 1. Cu(II) binding to ethylene diamine core poly(amidoamine) dendrimers in aqueous solutions. AB - This paper describes an investigation of the uptake of Cu(II) by poly(amidoamine) (PAMAM) dendrimers with an ethylenediamine (EDA) core in aqueous solutions. We use bench scale measurements of proton and metal ion binding to assess the effects of (i) metal ion-dendrimer loading, (ii) dendrimer generation/terminal group chemistry, and (iii) solution pH on the extent of binding of Cu(II) in aqueous solutions of EDA core PAMAM dendrimers with primary amine, succinamic acid, glycidol, and acetamide terminal groups. We employ extended X-ray absorption fine structure (EXAFS) spectroscopy to probe the structures of Cu(II) complexes with Gx-NH2 EDA core PAMAM dendrimers in aqueous solutions at pH 7.0. The overall results of the proton and metal ion binding measurements suggest that the uptake of Cu(II) by EDA core PAMAM dendrimers involves both the dendrimer tertiary amine and terminal groups. However, the extents of protonation of these groups control the ability of the dendrimers to bind Cu(II). Analysis of the EXAFS spectra suggests that Cu(II) forms octahedral complexes involving the tertiary amine groups of Gx-NH2 EDA core PAMAM dendrimers at pH 7.0. The central Cu(II) metal ion of each of these complexes appears to be coordinated to 2-4 dendrimer tertiary amine groups located in the equatorial plane and 2 axial water molecules. Finally, we combine the results of our experiments with literature data to formulate and evaluate a phenomenological model of Cu(II) uptake by Gx NH2 PAMAM dendrimers in aqueous solutions. At low metal ion-dendrimer loadings, the model provides a good fit of the measured extent of binding of Cu(II) in aqueous solutions of G4-NH2 and G5-NH2 PAMAM dendrimers at pH 7.0. PMID- 15835133 TI - Partitioning of small amphiphiles at surfactant bilayer/water interfaces: an avoided level crossing muon spin resonance study. AB - The temperature-dependent variation of local environment and reorientation dynamics of the small amphiphile 2-phenylethanol in lamellar phase dispersions of the dichain cationic surfactants, 2,3-diheptadecyl ester ethoxypropyl-1,1,1 trimethylammonium chloride (DHTAC) and dioctadecyldimethylammonium chloride (DODMAC), and the nonionic surfactant, tetra(ethylene glycol) n-dodecyl ether (C12E4), have been determined using avoided level crossing muon spin resonance spectroscopy (ALC-muSR). For cosurfactant radicals the hydrophobic or hydrophilic character of the surrounding media can be determined from their magnetic resonance signatures. Comparison of the three different bilayer-forming surfactant systems shows that the ALC-muSR technique is able to distinguish both major and subtle differences in the partitioning of the cosurfactant radicals between the different systems. PMID- 15835134 TI - Enhancement of Na+ diffusion in a bicontinuous cubic phase by the ionophore monensin. AB - Pulsed field gradient spin-echo NMR diffusion and relaxation measurements were used to investigate how the Na+ ionophore monensin affected the dynamics of sodium ions in a Myverol 18-99/saline bicontinuous Ia3d cubic phase (BCP). The monensin Na+ binding number was estimated from 23Na line widths to be between 3 and 6. The dependence of the apparent Na+ diffusion coefficient on the concentration of monensin revealed monensin-induced Na+ transport. At high monensin concentrations, the enhancement of D(Na+) was offset by Na+-monensin binding. The greatest enhancement was measured at short diffusion times (delta < or = 5 ms), which we explain in terms of the bicontinuous topology of the cubic phase and a combination of tortuosity and bilayer permeability effects. We also propose numerical simulations which would enable the separation of the two effects. To our knowledge, this is the first study of ionophore-mediated cation diffusion in a bicontinuous cubic phase. The approach could be used to study the dynamics of hydrophilic species in the aqueous channels of BCPs and similar structures, as well as to measure the ion-transporting efficiency of ionophores. PMID- 15835135 TI - Direct patterning of self-assembled monolayers on gold using a laser beam. AB - The development of a methodology to manipulate surface properties of a self assembled monolayer (SAM) of alkanethiol on a gold film using direct laser patterning is the objective of this paper. The present study demonstrates proof of the concept for the feasibility of laser patterning monolayers and outlines theoretical modeling of the process to predict the resulting feature size. This approach is unique in that it eliminates the need for photolithography, is noncontact, and can be extended to other systems such as SAMs on silicon wafers or potentially polymeric substrates. A homogeneous SAM made of 1-hexadecanethiol is formed on a 300-A sputtered film of gold (supported by a soda lime glass substrate). Localized regions are then desorbed by scanning the focal spot of a 488-nm continuous-wave argon ion laser beam under a nitrogen atmosphere. The desorption occurs as a result of a high substrate temperature produced by the moving laser beam with a Gaussian spatial profile at a constant speed of 200 microm/s. After completing the scans, the sample is dipped into a dilute solution of 16-mercaptohexadecanoic acid and a hydrophilic monolayer self-assembles along the previously irradiated regions. The resultant lines are viewed, and line widths are measured using both wetting with tridecane under a light microscope and scanning electron microscopy. Using the direct laser patterning method, we have produced straight line patterns with widths of 28-170 microm. A thermal model was constructed to predict the line width of the desorbed monolayer. The effect of the laser power, beam waist, and temperature dependence of the substrate conductivity on the theoretical predictions is considered. It is shown that the theoretical predictions are in good agreement with the experimental results, and, thus, the model can effectively be used to predict experimental results. PMID- 15835136 TI - Controlled rearrangement of adsorbed undecanol films on mica surfaces induced by an atomic force microscopy tip. AB - An undecanol film adsorbed on a mica surface was found to rearrange and spread in a position-controlled way induced by a tapping mode atomic force microscopy (AFM) probe. AFM images of varying scanning times showed that before forming an ordered monolayer the undecanol molecules were adsorbed on the mica surface in the disordered and disorganized status. With the proceeding of scanning, these undecanol molecules gradually formed an ordered and flat film. Such behavior was caused by the formation of a stable film and had never been reported for other alcohols. PMID- 15835137 TI - Design of new materials for methane storage. AB - One of the strategic goals of the modern automobile manufacturing industry is to replace gasoline and diesel with alternative fuels such as natural gas. In this report, we elucidate the desired characteristics of an optimal adsorbent for gas storage. The U.S. Department of Energy has outlined several requirements that adsorbents must fulfill for natural gas to become economically viable, with a key criterion being the amount adsorbed at 35 bar. We explore the adsorption characteristics of novel metal-organic materials (IRMOFs and molecular squares) and contrast them with the characteristics of two zeolites, MCM-41, and different carbon nanotubes. Using molecular simulations, we uncover the complex interplay of the factors influencing methane adsorption, especially the surface area, the capacity or free volume, the strength of the energetic interaction, and the pore size distribution. We also explain the extraordinary adsorption properties of IRMOF materials and propose new, not yet synthesized IRMOF structures with adsorption characteristics that are predicted to exceed the best experimental results to date by up to 36%. PMID- 15835138 TI - Adsorption of trinitrotoluene on uncoated silicon microcantilever surfaces. AB - We measured the adsorption characteristics of trinitrotoluene (TNT) on piezoresistive silicon microcantilever surfaces under ambient air using a well characterized TNT vapor generator. This allowed us to quantify the adsorption parameters and to estimate the sticking coefficient. The sticking coefficient initially increases with TNT exposure time and then levels off around 0.3. Atomic force microscopy images of silicon surfaces exposed to TNT revealed "island" formation of the adsorbate on the silicon surface. At low exposure times, mainly the number density of islands increased with exposure time; at longer exposure times, the size (in particular, height) of the islands grew, corresponding to the higher sticking coefficients. These observations can be qualitatively explained via the difference between TNT-surface and TNT-TNT interactions mediated by water molecules. PMID- 15835139 TI - Nanoscale aggregate structures of trisiloxane surfactants at the solid-liquid interface. AB - The self-associating structures at the solid-liquid interface of three nonionic trisiloxane surfactants ((CH3)3SiO)2Si(CH3)(CH2)3(OCH2CH2)n OH (n = 6, 8, and 12), or BEn, are studied as a function of substrate properties by atomic force microscopy (AFM) imaging and force measurement. These trisiloxane surfactants are known as superwetters, which promote rapid spreading of dilute aqueous solutions on low-energy surfaces. This study also attempts to relate the BEn surface aggregate structures at the solid-liquid interface to their superwetting behavior. Four substrates are used in the study: muscovite mica, highly oriented pyrolytic graphite, and oxidized silicon wafer with and without a full monolayer of self-assembled n-octadecyltrichlorosilane (OTS). The concentration of BEn is fixed at 2 times the critical aggregation concentration (CAC). The BEn surfactants are only weakly attracted to hydrophilic surfaces, more on oxidized silicon than on mica. All three form ordinary planar monolayers on HOPG and OTS covered oxidized silicon. The significance of surfactant adsorption on the AFM tip is investigated by comparing the force curves obtained by tips with and without thiol modification. The surface aggregate structures of the BEn surfactants correlate with their bulk structures and do not exhibit anomalous adsorption behavior. The adsorption behavior of the BEn superwetters is similar to that of the CmEn surfactants. Thus, our results confirm previous work showing that superwetting shares its main features with other classes of surfactants. PMID- 15835140 TI - Effects of surface hydrophobization on the growth of self-assembled monolayers on silicon. AB - The growth of self-assembled monolayers from octadecyltrichlorosilane (OTS) on modified silicon surfaces has been investigated. The influence of different immersion times in a deactivation reagent on the growth mechanism and the ordering of the films has been studied. Characterization of the films and the submonolayer coverage has been performed with tapping mode atomic force microscopy, ellipsometry, and infrared spectroscopy. We found that a deactivation of active sites led to a higher mobility of adsorbed molecules on the surface resulting in circular islands of highly ordered alkylsiloxane. However, upon prolonged immersion in OTS these ordered islands did not continue to grow and full monolayer coverage could not be obtained. Instead, an exchange reaction with the deactivation reagent leading to a disordered film between the ordered islands was observed. This was confirmed by external reflection infrared spectroscopy. PMID- 15835141 TI - In situ determination of the thermodynamic surface properties of chemically modified surfaces on a local scale: an attempt with the atomic force microscope. AB - We have monitored deflection-distance curves with an atomic force microscope (AFM) in contact mode, with a silicon nitride tip, on chemically modified silicon wafers, in the air. The wafers were modified on their surface by grafting self assembled monolayers (SAMs) of different functional groups such as methyl, ester, amine, or methyl fluoride. A chemically modified surface with a functionalized hydroxyl group was also considered. Qualitative analysis allowed us to compare adhesive forces versus chemical features and surface energy. The systematic calibration procedure of the AFM measurements was performed to produce quantitative data. Our results show that the experimentally determined adhesive force or thermodynamic work of adhesion increases linearly with the total surface energy determined with contact angles measured with different liquids. The influence of capillary condensation of atmospheric water vapor at the tip-sample interface on the measured forces is discussed. Quantitative assessment values were used to determine in situ the SAM-tip thermodynamic work of adhesion on a local scale, which have been found to be in good agreement with quoted values. Finally, the determination of the surface energy of the silicon wafer deduced from the thermodynamic work of adhesion is also proposed and compared with the theoretical value. PMID- 15835142 TI - Chiral morphologies and interfacial electronic structure of naphtho[2,3-a]pyrene on Au(111). AB - The adsorption of the two-dimensionally chiral naphtho[2,3-a]pyrene molecule has been studied on Au(111). Both structural and electronic properties of the naphtho[2,3-a]pyrene (NP)/Au(111) interface have been measured. Ultraviolet and X ray photoelectron spectroscopy have been employed to measure the energies of the molecular orbitals of the NP film with respect to the gold Fermi level. A Schottky junction with a large interface dipole (0.99 eV) is formed between Au(111) and NP. Temperature-programmed desorption was used to determine that adsorbed NP has a binding energy of 102.2 kJ/mol. Chiral domains have been observed with scanning tunneling microscopy due to the spontaneous phase separation of the 2-D enantiomers. Two distinct structural polymorphs have been observed, one of which has homochiral paired molecular rows. Models of the 2D structure are proposed that are in excellent agreement with experimental measurements. PMID- 15835143 TI - On the adsorption kinetics of surface-chemically pure n-dodecanoic acid at the air/water interface. AB - The dynamic surface tension data for n-dodecanoic acid in 0.005 M hydrochloric acid, for as-received as well as for surface-chemically pure solutions, show the presence of a prolonged induction period, clearly indicating that the adsorption of this nonionic surfactant is not simply diffusion-controlled. A kinetic model for the reversible formation of monolayer islands, long known in the field of electrochemistry, is shown to also apply to the adsorption of n-dodecanoic acid at the air/water interface. The rate constant increases linearly with increasing bulk concentration, while the induction time decreases exponentially. The phenomenon of nucleation at the air/water interface is consistent with the direct experimental observation of the formation of solid-like patches as the interfacial region is drastically compressed. PMID- 15835144 TI - Analysis of pseudopartial and partial wetting of various substrates by lead. AB - Lead drops exhibit partial wetting on some substrates and pseudopartial wetting on others. In pseudopartial wetting, a film is in equilibrium with a capillary body with a nonzero contact angle. Using a free energy formulation appropriate for the experiments, we show the conditions under which minimization of the system energy is accurately achieved by minimizing the energy of the film alone. Using a set of simple surface energy isotherms, we explain the various wetting behaviors of lead. We contrast isotherms for autophobing systems and the metallic systems considered here. PMID- 15835145 TI - Polyelectrolyte multilayer films of different charge density copolymers with synergistic nonelectrostatic interactions prepared by the layer-by-layer technique. AB - Random copolymers composed of diallyldimethylammonium chloride (DADMAC) and acrylamide with varying contents (8-100 mol %) of the cationic DADMAC component were alternated with polyanionic, fully charged poly(styrenesulfonate) to form multilayer thin films. UV-vis spectrophotometry, FTIR spectroscopy, and quartz crystal microgravimetry (QCM) were employed to follow multilayer buildup. Atomic force microscopy was used to obtain structural information. Layer thicknesses have been determined with small-angle X-ray scattering and ellipsometry, in addition to values calculated from QCM. While in previous work, a critical charge density limit could be observed, below which no layer growth is possible; in this system, multilayer formation takes place with copolymers with charge densities as low as 8 mol %. Instead of a continuous increase of adsorbed amounts with decreasing charge density above the critical charge density, as found in previous work, similar layer thicknesses for films with 100 and 8 mol % charged polyelectrolytes and maximally adsorbed amounts for copolymers in an intermediate charge density region have been found. This adsorption behavior is explained in terms of synergistic nonelectrostatic interactions between the polyelectrolytes used. PMID- 15835146 TI - Deposition and wetting characteristics of polyelectrolyte multilayers on plasma modified porous polyethylene. AB - Hydrophilic and chemically reactive porous media were prepared by adsorbing functional polymers at the surface of sintered polyethylene membranes. Modification of the membrane was accomplished by first exposing the membrane to an oxygen glow discharge gas plasma to introduce an electrostatic charge at the membrane surfaces. Cationic polyelectrolyte polyethylenimine (PEI) was adsorbed from solution to the anionic-charged surface to form an adsorbed monolayer. The adsorption of a second anionic polyelectrolyte onto the PEI layer allows further modification of the membrane surface to form a polyelectrolyte-bilayer complex. The conformation and stability of the adsorbed monolayers and bilayers comprising the modified surface are probed as a function of the polymer structure, charge density, and solubility. Using X-ray photoelectron spectroscopy analysis, we demonstrate that the presence of the polyelectrolyte multilayers drastically increases the density and specificity of the functional groups at the surface, more than what can be achieved through the plasma modification alone. Also, using the wicking rate of deionized, distilled water through the porous membrane to gauge the interfacial energy of the modified surface, we show that the membrane wicking rate can be controlled by varying the chemistry of the adsorbing polyelectrolytes and, to a lesser extent, by adjusting the polarity or ionic strength of the polyelectrolyte solution. PMID- 15835147 TI - Chemically transformable configurations of mercaptohexadecanoic acid self assembled monolayers adsorbed on Au(111). AB - Carboxyl-terminated self-assembled monolayers (SAMs) are commonly used in a variety of applications, with the assumption that the molecules form well-ordered monolayers. In this work, near-edge X-ray absorption fine structure measurements verify that well-ordered monolayers can be formed using acetic acid in the solvent. Disordered monolayers with unbound molecules present in the film result using only ethanol. A stark reorientation occurs upon deprotonation of the end group by rinsing in a KOH solution. This reorientation of the end group is reversible with tilted-over, hydrogen-bound carboxyl groups while the carboxylate ion end groups are upright. C(1s) photoemission shows that SAMs formed and rinsed with acetic acid in ethanol have protonated end groups, while SAMs formed without acetic acid have a large fraction of carboxylate-terminated molecules. PMID- 15835148 TI - Photocatalytic one-electron oxidation of biphenyl derivatives strongly coupled with the TiO2 surface. AB - One-electron oxidation of various biphenyl derivatives such as biphenyl (BP), 4 hydroxybiphenyl (HBP), 4,4'-biphenol (DHBP), 4-methoxy-4'-hydroxybiphenyl (MHBP), 4-cyano-4'-hydroxybiphenyl (CNHBP), 4-biphenylmethanol (BPM), and 4 biphenylethanol (BPE) adsorbed on the surface of TiO2 powder slurried in acetonitrile has been investigated by time-resolved diffuse reflectance spectroscopy. From the Langmuir adsorption isotherms for BP and HBP, it is suggested that the OH group plays an important role in adsorbing on the surface ofTiO2. The spectroscopic characteristics of charge transfer complexes of substrates and the TiO2 surface have been studied by steady-state diffuse reflectance spectroscopy. The high efficiency of a one-electron oxidation reaction was observed for hydroxyl-substituted BPs (HBP, DHBP, MHBP, CNHBP), compared with BPM and BPE, indicating that the strength of the electronic coupling element (H(DA)) between the electron acceptor and donor is a key factor in the one-electron oxidation of a substrate adsorbed on the TiO2 powder. The effects of the distance between the electron donor and the acceptor on the electronic coupling element are also discussed. PMID- 15835149 TI - Tilt of atomic force microscope cantilevers: effect on spring constant and adhesion measurements. AB - In atomic force microscopy, the cantilevers are mounted under a certain tilt angle alpha with respect to the sample surface. In this paper, we show that this increases the effective spring constant by typically 10-20%. The effective spring constant of a rectangular cantilever of length L can be obtained by dividing the measured spring constant by cos2 alpha(1 - 2D tan alpha/L). Here, alpha is the tilt angle and D is the size of the tip. In colloidal probe experiments, D has to be replaced by the radius of the attached particle. To determine the effect of tilt experimentally, the adhesion force between spherical borosilicate particles and planar silicon oxide surfaces was measured at tilt angles between 0 degrees and 35 degrees. The experiments revealed a significant decrease of the mean apparent adhesion force with a tilt of typically 20-30% at alpha = 20 degrees. In addition, they demonstrate that the adhesion depends drastically on the precise position of contact on the particle surface. PMID- 15835150 TI - Synthesis and supramolecular self-assembly study of a novel porphyrin molecule in Langmuir and Langmuir-Blodgett films. AB - The self-assembly and supramolecular engineering of porphyrins into ordered arrays have recently attracted much interest because of their promising application potential in molecular and electronic devices, spintronics, energy harvesting and storage, catalysis, and sensor development. We herein report the synthesis and supramolecular self-assembly study of a novel porphyrin molecule, 2Por-TAZ, in Langmuir and Langmuir-Blodgett films. The 2Por-TAZ molecule contains two porphyrin macrocycles attached to a triaminotriazine headgroup. Triaminotriazines are known to form a highly ordered linear supramolecular self assembly through complementary hydrogen bonding with barbituric acid molecules at the air-water interface. Surface pressure-area isotherm measurements and polarized UV-vis absorption spectroscopic studies indicate that the 2Por-TAZ molecules adopted an edge-on orientation at the air-water interface. Polarized UV vis absorption study also revealed that the 2Por-TAZ molecules formed linear supramolecular networks on pure water and barbituric acid subphase with porphyrin flat planes facing toward the compression direction. The binding of barbituric acid with 2Por-TAZ molecules was observed from the expansion of the Langmuir monolayer film. Compared to the transferred LB film from pure water subphase, both the UV-vis absorbance and fluorescence emission intensity of the LB film transferred from barbituric acid subphase increased significantly. PMID- 15835151 TI - Langmuir monolayers of bent-core molecules. AB - A systematic study of five different, symmetric bent-core liquid crystals in Langmuir thin films at the air/water interface is presented. Both the end chains (siloxane vs hydrocarbon) and the core (more or less amphiphilic) are varied, to allow an exploration of different possible layer structures at the interface. The characterization includes systematic surface pressure isotherms, Brewster angle microscopy, and surface potential measurements. The properties of these layers are strongly dependent on the individual type of molecule: the molecules with amphiphilic end chains lie quite flat on the surface, while the molecules with hydrophobic end chains construct multilayer structures. In both cases, the three dimensional collapse structure is reversible. PMID- 15835152 TI - SAMs of shape-persistent macrocycles: structure and binding on HOPG and Au(111). AB - Self-assembled monolayers of shape-persistent macrocycles have been adsorbed on highly oriented pyrolytic graphite and Au(111) substrates. Their structure and binding characteristics have been investigated by scanning tunneling microscopy, X-ray photoelectron spectroscopy, and ultraviolet photoelectron spectroscopy experiments. Special functionalization of the macrocycles for the first time has enabled their self-assembling and binding on a metal substrate and opened a new route for new functional nanostructures. PMID- 15835153 TI - Electrostatic free energy of weakly charged macromolecules in solution and intermacromolecular complexes consisting of oppositely charged polymers. AB - When oppositely charged polyelectrolytes are mixed in water, attraction between oppositely charged groups may lead to the formation of polyelectrolyte complexes (associative phase separation, complex coacervation, interpolymer complexes). Theory is presented to describe the electrostatic free energy change when ionizable (annealed) (macro-)molecules form a macroscopic polyelectrolyte complex. The electrostatic free energy includes an electric term as well as a chemical term that is related to the dissociation of the ionic groups in the polymer. An example calculation for complexation of polyacid with polybase uses a cylindrical diffuse double layer model for free polymer in solution and electroneutrality within the complex and calculates the free energy of the system when the polymer is in solution or in a polyelectrolyte complex. Combined with a term for the nonelectrostatic free energy change upon complexation, a theoretical stability diagram is constructed that relates pH, salt concentration, and mixing ratio, which is in qualitative agreement with an experimental diagram obtained by Bungenberg de Jong (1949) for complex coacervation of arabic gum and gelatin. The theory furthermore explains the increased tendency toward phase separation when the polymer becomes more strongly charged and suggests that complexation of polyacid or polybase with zwitterionic polymer (e.g., protein) of the same charge sign (at the "wrong side" of the iso-electric point) may be due (in part) to an induced charge reversal of the protein. PMID- 15835154 TI - Coadsorption of sodium dodecyl sulfate and a polyanion onto poly(ethylenimine) in multilayered thin films. AB - Mixed surfactant-polyelectrolyte multilayer films were fabricated by both ionic self-assembly and spin assembly. A polycation [PEI = poly(ethylenimine)] was deposited from a dilute solution, while a polyanion (PAZO = poly[1-[4-(3-carboxy 4-hydroxyphenylazo) benzenesulfonamido]-1,2-ethanediyl, sodium salt]) was deposited from a mixture containing a fixed concentration of polyanion and various concentrations of the anionic surfactant sodium dodecyl sulfate (SDS). Coadsorption of SDS and PAZO onto PEI layers was observed using both deposition methods and attributed to strong PEI-SDS interactions and entropic factors. Increasing the concentration of SDS resulted in films containing progressively less adsorbed PAZO. No further reduction in the amount of adsorbed PAZO was observed above the SDS critical micelle concentration. We attribute the film growth behavior to a fast adsorption of SDS onto PEI, followed by a slower adsorption of PAZO onto the remaining unoccupied binding sites. We observe that SDS interpenetrates throughout the PAZO and PEI layers, increasing the surface hydrophobicity of both. We observed similar behavior for both ionically self assembled and spin-assembled systems. PMID- 15835155 TI - Self-assembled monolayers of 4-mercaptopyridine on Au(111): a potential-induced phase transition in sulfuric acid solutions. AB - In situ scanning tunneling microscopy images of self-assembled monolayers (SAMs) of 4-mercaptopyridine (4-MPy) on Au(111) recorded in neat 0.1 M H2SO4 solutions provided evidence for a potential-induced phase transition over the range 0.40 0.15 V versus saturated calomel electrode. Analysis of the data was consistent with the presence of a (5 x square root(3)) and (10 x square root(3)) superstructure (phase A) at the positive end, that is, 0.40 V, for which the local coverage, theta(loc), is about 0.2 (two 4-MPy molecules per unit cell), which compresses at the negative end, that is, 0.15 V, to yield a much denser superstructure (phase B, theta(loc) ca. 0.5). This behavior is unlike that reported for the 4-MPy-Au(111) SAM prepared by identical means, in 0.1 M HClO4 (or in sulfate solutions of a much higher pH) for which only the (5 x square root(3)) superstructure was observed over the same potential range. The compression associated with the phase A to phase B transition is attributed to the formation of a hydrogen-bonded network of bisulfate coordinated in turn to the 4-MPy layer via the acidic hydrogens of the pyridinium moieties. Such conditions promote better packing of adsorbed 4-MPy species, which are aided by intermolecular pi-pi ring interactions, resulting in higher local coverages. PMID- 15835156 TI - On the existence of ordered organic adlayers at the Cu(111)/electrolyte interface. AB - We have reinvestigated the behavior of a Cu(111) electrode in pure and cinchonidine containing aqueous 0.1 M HClO4 solution by cyclic voltammetry (CV) and in situ electrochemical scanning tunneling microscopy (STM). In contrast to previous publications by Wan et al. (Langmuir 2000, 19, 1958-1962 and references cited therein) on Cu(111) in pure 0.1 M HClO4 which claimed an adsorbate-free Cu(111) surface in the entire potential range, we have found a highly ordered hexagonal adsorbate structure with a (4 x 4) unit cell, which is stable in the potential range from hydrogen evolution at -350 to -150 mV (RHE). The adsorbate free (1 x 1) Cu(111) surface is only visible in a fairly small potential range from -150 to +50 mV. A disordered surface structure is formed at more positive potentials which is interpreted by adsorption of an oxygen-containing species. Furthermore, the formation of a highly ordered cinchonidine adlayer on Cu(111) in 0.1 M HClO4 as reported by Wan et al. (J. Am. Chem. Soc. 2002, 124, 14300-14301) could not be reproduced here. In fact, the similarity of all structures reported by Wan et al. for a great variety of different organic adlayers on Cu(111) in HClO4 solution including cinchonidine with the (4 x 4) superstructure found here already in pure HClO4 solution (i.e., without organic solute) casts serious doubts on the validity of those previous results by Wan et al. in general. PMID- 15835157 TI - Operation of the quartz crystal microbalance in liquids: derivation of the elastic compliance of a film from the ratio of bandwidth shift and frequency shift. AB - For a thin viscoelastic film deposited on a quartz crystal microbalance in a liquid environment, the change in dissipation induced by the presence of a film is proportional to the film's elastic compliance, Jf'. This surprising result is a consequence of the fact that the film is "clamped" by a viscous fluid. PMID- 15835158 TI - Frequency-dependent electromechanics of aqueous liquids: electrowetting and dielectrophoresis. AB - Electrowetting on dielectric and dielectrophoretic electromechanical mechanisms dominate microfluidic actuation in the low- and high-frequency limits, respectively. The frequency-dependent relationship between these two mechanisms has been clarified by the Maxwell stress tensor and a simple RC circuit model. In this paper, we report extensive height-of-rise measurements obtained with vertical, parallel, dielectrically coated electrodes to test this relationship using deionized water and solutions containing sugar and salt. For DC and AC (20 Hz to 20 kHz) voltage magnitudes up to approximately 150 V-rms, the data are highly reproducible and, within experimental error, consistent with the square law predictions of the model. Eventually as voltage is increased, a saturation phenomenon is observed which exhibits a weak dependence on frequency and is probably correlated to contact angle saturation. PMID- 15835159 TI - Analysis of myoglobin adsorption to Cu(II)-IDA and Ni(II)-IDA functionalized Langmuir monolayers by grazing incidence neutron and X-ray techniques. AB - The adsorption of myoglobin to Langmuir monolayers of a metal-chelating lipid in crystalline phase was studied using neutron and X-ray reflectivity (NR and XR) and grazing incidence X-ray diffraction (GIXD). In this system, adsorption is due to the interaction between chelated divalent copper or nickel ions and the histidine moieties at the outer surface of the protein. The binding interaction of histidine with the Ni-IDA complex is known to be much weaker than that with Cu IDA. Adsorption was examined under conditions of constant surface area with an initial pressure of 40 mN/m. After approximately 12 h little further change in reflectivity was detected, although the surface pressure continued to slowly increase. For chelated Cu2+ ions, the adsorbed layer structure in the final state was examined for bulk myoglobin concentrations of 0.10 and 10 microM. For the case of 10 microM, the final layer thickness was approximately 43 A. This corresponds well to the two thicker dimensions of myoglobin in the native state (44 A x 44 A x 25 A) and so is consistent with an end-on orientation for this disk-shaped protein at high packing density. However, the final average volume fraction of amino acid segments in the layer was 0.55, which is substantially greater than the value of 0.44 calculated for a completed monolayer from the crystal structure. This suggests an alternative interpretation based on denaturation. GIXD was used to follow the effect of protein binding on the crystalline packing of the lipids and to check for crystallinity within the layer of adsorbed myoglobin. Despite the strong adsorption of myoglobin, very little change was observed in the structure of the DSIDA film. There was no direct evidence in the XR or GIXD for peptide insertion into the lipid tail region. Also, no evidence for in-plane crystallinity within the adsorbed layer of myoglobin was observed. For 0.1 microM bulk myoglobin concentration, the average segment volume fraction was only 0.13 and the layer thickness was < or = 25 A. Adsorption of myoglobin to DSIDA-loaded with Ni2+ was examined at bulk concentrations of 10 and 50 microM. At 10 microM myoglobin, the adsorbed amount was comparable to that obtained for adsorption to Cu2+-loaded DSIDA monolayers at 0.1 M. But interestingly, the adsorbed layer thickness was 38 A, substantially greater than that obtained at low coverage with Cu-IDA. This indicates that either there are different preferred orientations for isolated myoglobin molecules adsorbed to Cu-IDA and Ni-IDA monolayer films or else myoglobin denatures to a different extent in the two cases. Either interpretation can be explained by the very different binding energies for individual interactions in the two cases. At 50 microM myoglobin, the thickness and segement volume fraction in the adsorbed layer for Ni-IDA were comparable to the values obtained with Cu IDA at 10 microM myoglobin. PMID- 15835160 TI - Contact angle hysteresis, adhesion, and marine biofouling. AB - Adhesive and marine biofouling release properties of coatings containing surface oriented perfluoroalkyl groups were investigated. These coatings were prepared by cross-linking a copolymer of 1H,1H,2H,2H-heptadecafluorodecyl acrylate and acrylic acid with a copolymer of poly(2-isopropenyl-2-oxazoline) and methyl methacrylate at different molar ratios. The relationships between contact angle, contact angle hysteresis, adhesion, and marine biofouling were studied. Adhesion was determined by peel tests using pressure-sensitive adhesives. The chemical nature of the surfaces was studied by using X-ray photoelectron spectroscopy. Resistance to marine biofouling of an optimized coating was studied by immersion in seawater and compared to previous, less optimized coatings. The adhesive release properties of the coatings did not correlate well with the surface energies of the coatings estimated from the static and advancing contact angles nor with the amount of fluorine present on the surface. The adhesive properties of the surfaces, however, show a correlation with water receding contact angles and contact angle hysteresis (or wetting hysteresis) resulting from surface penetration and surface reconstruction. Coatings having the best release properties had both the highest cross-link density and the lowest contact angle hysteresis. An optimized coating exhibited unprecedented resistance to marine biofouling. Water contact angle hysteresis appears to correlate with marine biofouling resistance. PMID- 15835161 TI - Structural characteristics of activated carbons and ibuprofen adsorption affected by bovine serum albumin. AB - Structural characteristics of a series of MAST carbons were studied using scanning electron microscopy images and the nitrogen adsorption isotherms analyzed with several models of pores and different adsorption equations. A developed model of pores as a mixture of gaps between spherical nanoparticles and slitlike pores was found appropriate for MAST carbons. Adsorption of ibuprofen [2 (4-isobutylphenyl)propionic acid] on activated carbons possessing different pore size distributions in protein-free and bovine serum albumin (BSA)-containing aqueous solutions reveals the importance of the contribution of mesopores to the total porosity of adsorbents. The influence of the mesoporosity increases when considering the removal of the drug from the protein-containing solution. Cellulose-coated microporous carbon Norit RBX adsorbs significantly smaller amounts of ibuprofen than uncoated micro/mesoporous MAST carbons whose adsorption capability increases with increasing mesoporosity and specific surface area, burnoff dependent variable. A similar effect of broad pores is observed on adsorption of fibrinogen on the same carbons. Analysis of the ibuprofen adsorption data using Langmuir and D'Arcy-Watt equations as the kernel of the Fredholm integral equation shows that the nonuniformity of ibuprofen adsorption complexes diminishes with the presence of BSA. This effect may be explained by a partial adsorption of ibuprofen onto protein molecules immobilized on carbon particles and blocking of a portion of narrow pores. PMID- 15835162 TI - Effect of the mobility of ligands in polyrotaxanes on order structure of water clusters. AB - The effect of the mobility of ligands (maltose groups) in the polyrotaxanes (pRXs) on the structure of the surrounding water molecules was investigated. Raman spectra of collective OH stretching vibration of water molecules in aqueous solutions of maltose-pRX conjugates with different alpha-cyclodextrin (alpha-CD) threading on a poly(ethylene glycol) (PEG) chain was measured. The mobility of maltose groups was estimated by measuring the relaxation time T2 of the C1 protons in maltose groups bound on alpha-CD by NMR experiment. A positive correlation between the Raman intensity of the collective band and the relaxation time T2 was obtained. This result indicates that the degree of order of the water clusters is higher as the mobility of maltose groups increases in these conjugate solutions. It is suggested that rapid motion of maltose groups in the pRX conjugate can contribute to preserving ordered structure of the bulk water clusters. PMID- 15835163 TI - Synthesis of ZnO nanoparticles on a clay mineral surface in dimethyl sulfoxide medium. AB - Nanocrystalline ZnO particles have been prepared with different methods using zinc cyclohexanebutyrate as precursor in dimethyl sulfoxide (DMSO) medium via alkaline hydrolysis. A series of preparations were carried out in the presence of layered silicates (kaolinite and montmorillonite). It was revealed by different measurement techniques that the presence of the clay minerals has a stabilization influence on the size of the ZnO nanocrystals. UV-vis absorption spectra show a blue shift when the nanoparticles are prepared in the presence of the clay minerals. The average particle diameters calculated from the Brus equation ranged from 2.6 to 13.0 nm. The UV-vis spectra of the synthesized nanoparticles did not show any red shift after 2-3 days, demonstrating that stable ZnO nanocrystals are present in the dispersions. The presence of the ZnO nanoparticles was also proven by fluorescence measurements. A number of the nanoparticles are incorporated into the interlamellar space of the clays, and an intercalated structure is formed as proven by X-ray diffraction (XRD) measurements. The size of the nanoparticles in the interlamellar space is in the range of 1-2 nm according to the XRD patterns. Transmission electron microscopy and high-resolution transmission electron microscopy investigations were applied to determine directly the particle size and the size distribution of the nanoparticles. PMID- 15835164 TI - Development of novel pillared clays for the encapsulation of inorganic complexes. AB - Several pillared clays were prepared by using a polyalcohol (ethylene glycol or poly(vinyl alcohol)) or a poly(ethylene oxide) surfactant as an interlayer gallery template and an aluminum oligomer species as the pillaring agent. The use of polyalcohols or nonionic surfactants, such as Tergitol, gave materials which, in general, presented larger basal spacing than those found for the solids prepared by a similar procedure but without additives. The initial positive effect in the expansion of the clay interlayers was not totally retained after calcination of the materials; most probably, at the end, the basal spacing is still ruled by the intercalating aluminum species. The pillared clay with the largest basal spacing and specific surface area was used to encapsulate copper(II) complexes with pentadentate N3O2 Schiff base ligands derived from copper(II) acetylacetonate by in situ synthesis. The characterization made (X-ray diffraction, X-ray photoelectron spectroscopy, FTIR spectroscopy, chemical analysis, and low-temperature N2 adsorption) provided evidence that copper(II) complexes with pentadentate N3O2 Schiff base ligands were efficiently entrapped within the lower dimension pores of the pillared clay and that they interact strongly with the pillared clay matrix. PMID- 15835165 TI - Polymer-stabilized gold nanoparticles with high grafting densities. AB - A series of polymer-coated Au nanoparticles have been prepared using the "grafting-to" approach. Thiol-terminated polystyrene and poly(ethylene oxide) ligands are found to form dense brushes on the faceted gold nanoparticle surfaces. Depending on the polymer, the ligand grafting densities on the gold nanoparticles are 1.2- to 23.5-fold greater than those available via self assembled monolayer formation of the corresponding two-dimensional gold surfaces. PMID- 15835166 TI - Effect of pH and salt on the stiffness of polyelectrolyte multilayer microcapsules. AB - By using a combination of atomic force and confocal microscopy, we explore the effect of pH and salt on the stiffness of polyelectrolyte microcapsules with shells composed of strong polyanions and weak polycations. The stiffness of the capsules was found to be largest in water. It decreases slightly with added salt and gets much smaller both in acidic and in alkaline solutions. The moderate softening of the capsules in electrolyte solutions indicates that even high salt concentration does not significantly dissociate polyelectrolytes in the multilayer. The dramatic softening of the capsules at high pH probably reflects a decrease in the charge density of a polycation, which leads to a reduction in the number of ionic cross-links. In contrast, low stiffness of the capsules in acidic solutions seems to be connected mostly with the enhanced permeability of the multilayer shell. PMID- 15835167 TI - Novel coassembly route to Cu-SiO2 MCM-41-like mesoporous materials. AB - A series of mesostructured Cu-SiO2 composites have been synthesized with sodium metasilicate (Na2SiO3) and cuprammonia nitrate (Cu(NH3)4(NO3)2) respectively used as Si and Cu sources. The synthetic procedures were conducted at room temperature, and cetyltrimethylammonia bromide was used as a template. Under our experimental conditions, ordered mesoporous Cu-SiO2 composites could be obtained with a copper content up to 16.8 wt %. Average pore diameters (2.80-3.15 nm), wall thickness (1.30-2.20 nm), and specific surface area (1020-690 m2/g) are found to vary linearly with copper content (0-16.8 wt %). Results of thermal gravimetry-differential thermal analysis reveal the collapse temperature of the order structure starts at approximately 1250 K for mesoporous Cu-SiO2 with 16.8 wt % copper content. As indicated by the outcomes of inductively coupled plasma and X-ray photoelectron spectroscopy studies, copper is mainly incorporated inside the pore wall rather than embedded on the wall surface. Copper species strongly interact with silica, and calcination at high temperatures cannot cause phase separation between silica and copper oxide. Cu status in mesoporous Cu-SiO2 composites is similar to that in copper silicate in neighboring structures. Based on the results, a S+ I- I+ I- mechanism is proposed in which copper entities are surrounded by silicon species during synthesis of the mesostructured composite. PMID- 15835168 TI - Characterization of structural changes of poly(vinyl methyl ether) gamma irradiated in diluted aqueous solutions. AB - An aqueous deoxygenated dilute PVME solution (cP = 0.5 mass %) was irradiated with gamma-rays at different radiation doses (D = 0.2-10 kGy). At these concentrations and doses contracted molecules are formed but no macroscopic networks. For the analysis of the structural changes, the irradiated samples were analyzed by size exclusion chromatography (SEC) with a triple detector system, static light scattering, and viscometry. SEC measurements necessitate for the application of the universal calibration principle the knowledge of the Kuhn-Mark Houwink (KMH) parameters. To obtain these parameters commercially available poly(vinyl methyl ether) was fractionated and analyzed by the same means. We found at 30 degrees C a KMH relationship to [eta] = 0.0226 (mL/g) x Mw(0.67) in THF. We observed an increase in molar masses without any significant structural changing for D < 0.5 kGy. With increasing radiation dose, the intramolecular cross-linking reaction becomes more and more important. The results of viscosity measurements show a slight increase in contraction for 1.0 kGy. For irradiation dose higher than D > 1.5 kGy, a strong contraction occurs. For D > 5.0 kGy, the favored intramolecular reactions lead to the formation of microgels. PMID- 15835169 TI - Near-critical CO2 in mesoporous silica studied by in situ FTIR spectroscopy. AB - Attenuated total reflection Fourier transform infrared spectroscopy was used to correlate the band shift of the nu2 vibrational band of carbon dioxide with the density of the fluid. Upon adsorption of CO2 on mesoporous silica and a nonporous SiO2 film, additional bands were detected due to interactions of CO2 with SiO2. Near the saturation pressure for the porous samples, the absorbance of the nu2 band increased strongly, which was concluded to be caused by liquidlike CO2 inside the pores. Integration of single-beam-sample-reference spectra between bulk CO2 and CO2 adsorbing on the mesoporous silica coated on one part of the internal reflection element revealed excess adsorption type isotherms with sharp maxima at 21 degrees C. A flatter curve shape could be observed at 25 degrees C, which allowed estimating the pore critical temperature. Moreover, the density of the fluid inside and outside the pores could be compared. Over the investigated ranges of pressure, temperature, and pore size, the results evidenced that the CO2 density was always higher in the silica pores than in the bulk, even under supercritical conditions. This has important consequences on the pressure dependence of dissolution power and diffusivity of fluids in mesoporous solids. An overview is given on the influences of fluid phase behavior in the bulk and in the pores at various conditions on solubility and diffusivity. PMID- 15835170 TI - Synthesis and structure of nanocrystalline TiO2 with lower band gap showing high photocatalytic activity. AB - Nanocrystalline TiO2 was synthesized by the solution combustion method using titanyl nitrate and various fuels such as glycine, hexamethylenetetramine, and oxalyldihydrazide. These catalysts are active under visible light, have optical absorption wavelengths below 600 nm, and show superior photocatalytic activity for the degradation of methylene blue and phenol under UV and solar conditions compared to commercial TiO2, Degussa P-25. The higher photocatalytic activity is attributed to the structure of the catalyst. Various studies such as X-ray diffraction, Raman spectroscopy, Brunauer-Emmett-Teller surface area, thermogravimetric-differential thermal analysis, FT-IR spectroscopy, NMR, UV-vis spectroscopy, and surface acidity measurements were conducted. It was concluded that the primary factor for the enhanced activity of combustion-synthesized catalyst is a larger amount of surface hydroxyl groups and a lowered band gap. The lower band gap can be attributed to the carbon inclusion into the TiO2 giving TiO(2-2x)C(x) VO2**. PMID- 15835171 TI - Postsynthesis stabilization of free-standing mesoporous silica films. AB - Mixed ammonia-water vapor postsynthesis treatment provides a simple and convenient method for stabilizing mesostructured silica films. X-ray diffraction, transmission electron microscopy, nitrogen adsorption/desorption, and solid-state NMR (13C, 29Si) were applied to study the effects of mixed ammonia-water vapor at 90 degrees C on the mesostructure of the films. An increased cross-linking of the silica network was observed. Subsequent calcination of the silica films was seen to cause a bimodal pore-size distribution, with an accompanying increase in the volume and surface area ratios of the primary (d = 3 nm) to secondary (d = 5-30 nm) pores. Additionally, mixed ammonia-water treatment was observed to cause a narrowing of the primary pore-size distribution. These findings have implications for thin film based applications and devices, such as sensors, membranes, or surfaces for heterogeneous catalysis. PMID- 15835172 TI - Synthesis, characterization, and surface immobilization of platinum and palladium nanoparticles encapsulated within amine-terminated poly(amidoamine) dendrimers. AB - Platinum and palladium dendrimer-encapsulated nanoparticles (DENs) were prepared within commercially available, fourth-generation, amine-terminated, poly(amidoamine) dendrimers (G4-NH2). The synthesis is carried out by selectively encapsulating metal complexes within the dendrimer and then reducing the resulting composite. Intradendrimer complexation requires control over the solution pH to prevent attachment of the metal complexes to primary amine groups on the dendrimer periphery. That is, the surface primary amines of the dendrimer must be selectively protonated in the presence of the interior tertiary amines. The metal-ion encapsulation and reduction processes were characterized by UV-vis spectroscopy. Forty-atom Pt and Pd DENs were examined by high-resolution transmission electron microscopy, which showed that the mean particle sizes were 1.4 and 1.5 nm, respectively, and that both were nearly monodisperse (standard deviation = 0.3 nm). The free amine groups on the dendrimer surface were used to link Pd DENs to monolithic Au surfaces via an intermediate self-assembled monolayer adhesion layer. PMID- 15835173 TI - Fabrication of composite films containing zirconia and cationic polyelectrolytes. AB - Composite films were prepared by electrophoretic deposition of poly(ethylenimine) or poly(allylamine hydrochloride) combined with cathodic precipitation of zirconia. Films of up to several micrometers thick were obtained on Ni, Pt, stainless-steel, graphite, and carbon-felt substrates. When the concentration of polyelectrolytes in solutions and the deposition time were varied, the amount of the deposited material and its composition can be varied. The electrochemical intercalation of yttria-stabilized zirconia particles into the composite films has been demonstrated. Obtained results pave the way for the electrodeposition of other polymer-ceramic composites. The deposits were studied by thermogravimetric analysis, X-ray diffraction analysis, scanning electron microscopy, and atomic force microscopy. The mechanisms of deposition are discussed. PMID- 15835174 TI - Dynamic alterations of fibronectin layers on copolymer substrates with graded physicochemical characteristics. AB - Desorption and exchange of preadsorbed fibronectin layers in pure buffer solution and solutions of human serum albumin or fibronectin, respectively, were studied in dependence on the physicochemical characteristics of maleic acid copolymer films used as substrates. Although the preadsorbed amount of fibronectin differed only slightly, the protein was found to exhibit a significantly enhanced anchorage at the more hydrophobic polymer surface as compared to the more hydrophilic and more negatively charged polymer surface. The preadsorbed fibronectin layer was most efficiently exchanged by fibronectin (i.e., in the homodisplacement process) while pure buffer solution and human serum albumin solutions induced desorption or exchange of fibronectin to lower and similar degrees. An increase of the total adsorbed amount of protein due to additional adsorption of fibronectin or human serum albumin accompanied the partial exchange of the preadsorbed fibronectin in the displacement experiments. Evaluation of the kinetics of desorption and exchange of fibronectin at any of the substrates revealed two kinds of surface-attached protein populations--a fast desorbing species and a species with a slow desorption and exchange rate. By a multivariate regression analysis the surface characteristics of the polymer substrate were confirmed to determine the degree of protein desorption and exchange while the dynamics of the layer alteration was found to solely depend on the diffusion behavior of the proteins. PMID- 15835175 TI - Property influence of polyanilines on photovoltaic behaviors of dye-sensitized solar cells. AB - The influence of polyanilines (PANIs) as hole conductors on the photovoltaic behaviors of dye-sensitized solar cells is studied. The current-voltage (I-V) characteristics and the incident photon to current conversion efficiency (IPCE) curves of the devices are determined as the function of different conductivities and morphologies of PANIs. The results show that the conductivity of PANIs affects the performance of the devices greatly, and PANI with the intermediate conductivity value (3.5 S/cm) is optimum. In addition, the effects of both the film formation property and the cluster size of polyanilines on the photovoltaic behaviors of the devices are also discussed. PMID- 15835176 TI - Linear free energy relationships between dissolution rates and molecular modeling energies of rhombohedral carbonates. AB - Bulk and surface energies are calculated for endmembers of the isostructural rhombohedral carbonate mineral family, including Ca, Cd, Co, Fe, Mg, Mn, Ni, and Zn compositions. The calculations for the bulk agree with the densities, bond distances, bond angles, and lattice enthalpies reported in the literature. The calculated energies also correlate with measured dissolution rates: the lattice energies show a log-linear relationship to the macroscopic dissolution rates at circumneutral pH. Moreover, the energies of ion pairs translated along surface steps are calculated and found to predict experimentally observed microscopic step retreat velocities. Finally, pit formation excess energies decrease with increasing pit size, which is consistent with the nonlinear dissolution kinetics hypothesized for the initial stages of pit formation. PMID- 15835177 TI - Role of capillary stresses in film formation. AB - Stresses generated during film formation were deduced from the deflection of a copper cantilever coated with a drying latex. Experiments with particles of varying radii and glass transition temperatures (Tg) focused on conditions for which capillary stresses normal to the film deform the particles to close the voids. Soft particles (low Tg) formed continuous films, but hard ones (high Tg) produced fascinating arrays of cracks. For both soft and rigid particles, the lateral stresses were tensile and scaled on the surface tension divided by the particle radius. Clearly, tensile stresses in the plane of the film responsible for cracking arise from the same capillary pressure that drives compression in the normal direction. Solving the model (Routh & Russel 1996, 1999) for lateral flow of the fluid dispersion prior to close packing and deformation of the solid beyond close packing yields volume fraction, film thickness, and stress profiles for comparison with observations for both film-forming and film-cracking cases. PMID- 15835178 TI - Ion exchange effects on the electrical conductivity of acidified (HCl) sodium dodecyl sulfate solutions. PMID- 15835179 TI - PAMAM dendrimer-induced cross-linking modification of polyimide membranes. PMID- 15835180 TI - Surface phase partitioning in film formation of waterborne polyurethanes. Monte Carlo simulations and internal-reflectance IR imaging. PMID- 15835181 TI - On the critical radius in Ostwald ripening. PMID- 15835182 TI - Initial spreading dynamics of supported lipid monolayers. PMID- 15835183 TI - Hierarchical texture of calcium carbonate crystals grown on a polymerized Langmuir-Blodgett film. PMID- 15835184 TI - Structure and dynamics of cationic surfactants intercalated in synthetic clays. PMID- 15835185 TI - Standard states and activity coefficients of interfacial species. PMID- 15835186 TI - Energy transfer in fluorescent silica nanoparticles. PMID- 15835187 TI - Small-angle neutron scattering study of the structure of superswollen micelles formed by a highly asymmetric poly(oxybutylene)-poly(oxyethylene) diblock copolymer in aqueous solution. PMID- 15835188 TI - ONW carries out initiatives to tackle Mississippi's Nursing Workforce issues. PMID- 15835189 TI - 2005 legislative bills related to health care and/or nursing. PMID- 15835190 TI - Who's taking care of the old folks? Health care for older adults. PMID- 15835191 TI - Pediatric trauma and initial stabilization. PMID- 15835192 TI - Pediatric acute severe asthma. PMID- 15835193 TI - Life-threatening pediatric poisonings. AB - Initial therapy of the poisoned child should follow the basic principles of advanced life support and include support of a patent airway and adequate oxygenation, ventilation, and circulation. Ingestion of certain substances require agent-specific therapy which may modify the standard resuscitative approach. Children can present following unknown or unsuspected ingestions. Any child with unexplained altered level of consciousness, respiratory, circulatory derangement, seizures or metabolic abnormalities should be considered for toxic ingestion. Knowing the typical signs and symptoms of the common toxic syndromes can help identification of the unknown ingestion. Recommendations for decontamination, detoxification, and antidotal therapies have changed over the past 10 years. Clinicians should be aware of the community resources available to help guide the evaluation and management of the poisoned child. PMID- 15835194 TI - Pediatric diabetic ketoacidosis. PMID- 15835195 TI - Office preparedness for pediatric emergencies. PMID- 15835196 TI - Thermal injury in children. PMID- 15835197 TI - Emergency management of status epilepticus in children. PMID- 15835198 TI - Nursing education. PMID- 15835199 TI - Government affairs. PMID- 15835200 TI - Magnet status--first in Tennessee: Johnson City Medical Center. PMID- 15835201 TI - Ageism, elitism, and anti-intellectualism in nursing. PMID- 15835203 TI - Clinical nursing in adult epilepsy. AB - In this issue, we profile the role of nurses, who are the helping hands, listeners and educators who teach the world about epilepsy. They work on the front line of epilepsy health care. The following articles describe their roles, experiences and invaluable contribution to patients with epilepsy and their families. PMID- 15835202 TI - Writing proposals for research funds. AB - In this paper, we provide generic guidelines for writing proposals for research funds. These guidelines apply to quantitative, qualitative and mixed methods research proposals. A proposal for research funding provides an overview of the planned research and is required by funding agencies. The proposal is submitted for evaluation and ranking by a review committee according to the criteria put forth by that particular agency. It may be necessary to submit proposals to more than one agency to acquire sufficient operating and infrastructure funding for a research project. PMID- 15835204 TI - Kansas rural health networks: collaborating to improve health care. PMID- 15835205 TI - The Kansas Rural Health Options Project: providing assistance to rural providers. PMID- 15835206 TI - What is FLEX and how does it serve Kansas? Kansas capitalizes on unique federal grant program. PMID- 15835207 TI - Kansas nursing occupational outlook 2000-2010. PMID- 15835208 TI - 2004 Kansas Nursing Wage Survey. PMID- 15835209 TI - Self-administration of medication--S.B. 10. As amended by Senate Committee on Public Health and Welfare. PMID- 15835210 TI - [Health and territories: local policies' stakes]. PMID- 15835211 TI - [A breath for fresh air...with respect to a dictionary for medical thoughts]. PMID- 15835212 TI - [The state of affairs prior to the Regional Health Education Plan in the Nord/ Pas-de-Calais region: the present case, facts and expectations]. AB - The work on the development of the regional health education plan (SREPS) in the Nord/Pas-de-Calais region has been analysed and viewed in the light of the specific regional context. A number of statements can now be made on this work, as it underlines the expectations of the different health education professionals. The article testifies to the fact that the implementation of the SREPS, through its dynamic approach, contributes to the creation and reinforcement of a common culture while uncovering the road which remains left to travel. The improved situation allows for a more effective comparison with other regional and national situations. PMID- 15835213 TI - [Acting in the face of uncertainty: the decision to stop life support in the case of neo-natal resuscitation]. AB - This paper examines the way in which paediatricians manage in the case of neo natal resuscitation to make the decision to continue or stop the procedures, and this within a context characterised by the lack of reliability on scientific norms. The authors aimed to highlight that which orients the medical decisions in a local context of uncertainty. The data have been extracted from a significantly sized field project (based on observations and interviews) carried out in two neo natal intensive care service units. PMID- 15835214 TI - [Self-evaluation of the quality of life of children aged 6 to 12 years old: analysis of the concept and development of a prototype tool]. AB - The objective of this work was to assess the quality of life of children aged 6 to 12 years old. The authors present the different conceptual and operational steps which lead to the construction of a prototype tool. It was a generic tool composed of 63 items which covered the classically described areas of life. Its originality was due to the method of construction used which included children's involvement, to the pictorial representation of real-life situations for children, and to its adaptation for gender. The evaluation of this tool showed a good level of acceptability but statistical analysis revealed some metrological inadequacies leading to the reconsideration of the initial concepts and their methods of exploration. Therefore a new tool was constructed whose validation is presented in a second article in the next issue. PMID- 15835215 TI - [Determining students health information needs at the Service of Inter-university Preventative Medicine in Bordeaux]. AB - At the request of the Service of Academic Preventative Medicine (APM) in Bordeaux, this study was carried out in the year 2000 and aimed to identify the health information needs of the university's student population. The questionnaire, which was completed by the students themselves, mainly explored health information needs, health status and the students' living conditions. Of the 370 total students coming from all University disciplines who participated in this study, a majority of them were women (61%), with the median age of 21 years old. The majority of students (69%) said that they were stressed and/or depressed and more than one-third were tired. Nearly 20% of the students were smokers, 9% declared that they regularly drank alcohol and 7% of them reported that they never consulted a physician. When asked specifically about their health information needs, they expressed a preference for receiving information on sexually transmitted diseases, how to stay in good health and on issues like psychology, depression and suicide. These themes differed according to gender and risk behaviour. These results permitted a better understanding of the students' health information needs and provided the knowledge necessaryin order to streamline and appropriately adapt the APM's activities for the following academic year in order to be more focused to respond to these needs. PMID- 15835216 TI - [Patients and clients integrated in the process of constructing the regional health organisation plans]. AB - The government orders of April 1996 were devoted to the second generation Regional Health Organisation Plans (SROS II) as health planning levers. One of the main issues at stake in their development was the need to favour an overall management of the regional public health priorities and to involve the system's clients in them. In order to better understand the opinions of the clients themselves and the opinions of the health planning professionals on the participation of the clients in SROS II, a national sample was selected in order to take into account the diversity of regional experiences, the different institutional linkages and the unity of both the place and the problem at hand. Two series of 42 interviews conducted at 6 month intervals were recorded and a thematic analysis according to a validated interview grid was carried out by the Analytical Laboratory for Social and Health Policy (LAPSS) at the National School of Public Health (ENSP). This survey shows that the first attempt to include the participation of the system's clients in the health planning efforts by the SROS II was appreciated and recognised as legitimate and useful, yet suffered from a lack of legibility and of a clear definition of its role. Other participation mechanisms remain to be investigated and explored as they could be enlarged in the next plan, in which the clients could become a separate entity of actors as an entirely independent group. PMID- 15835217 TI - [Doctors' opinions on palliative care: results from a French survey on physicians attitudes and practices]. AB - OBJECTIVE: To examine practitioners' opinions on palliative care. METHOD: Statistical analyses were carried out on data collected from 1000 physicians recruited to participate in a study on knowledge, attitudes and practices with respect to palliative care ("Palliative Care 2002"). A cluster analysis (CAH) was conducted based on responses to nine questions in order to identify homogeneous and contrasting profiles of opinions on palliative care. RESULTS: Two distinctly different categories of profiles emerged. The first one had a restrictive approach to palliative care (n = 387; 38.7%), while the second one supported its expansion (n = 613; 61.3%). CONCLUSION: A newly revised communication campaign which favoured and recognised palliative care could assist in the dissemination of related practices among physicians. PMID- 15835218 TI - [Deficiencies in treating pain of elderly people suffering from a severe cognitive impairment]. AB - The objective of this study was to verify if the provision of care aimed at reducing pain was identical between elderly people suffering from a server cognitive distortion and other elderly people. The sample, taken from a two tiered survey, was composed of 352 elderly people living in 34 retirement homes or other similar establishments housing the elderly. We categorised the residents into 2 groups: Group I was comprised of those who had a complete loss of physical autonomy, and Group II contained all of the others. Within these two groups, we compared four types of medicine of which the prescription was given at the request of the patient following a complaint, and four types of medicine prescribed as a result of the analysis of biological tests and screening and not as a result of a complaint from the patient. The results showed that the elderly people having a severe cognitive impairment received significantly less medication aimed at reducing pain than the other elderly people. This difference is not recognised when the medication is prescribed as a result of a need noted due to a biological check-up and analysis. PMID- 15835219 TI - [Evaluation of a medical teaching programme in public health in Laos]. AB - Specialised medical training is progressively being established in Laos at the request of the Ministry of Health. A medical teaching programme in public health has been organised by the Cooperation Committee with Laos (CCL) in collaboration with the National Institute for Public Health (INSP) and has been taking place since 1997 within the framework of Franco-Laos cooperation. At the end of the validation of the second promotion of senior medical management staff (2002-2004) and of the presentation of the first Masters in Public Health in Laos, an evaluation of this training programme was carried out with the aim of completely transferring the project over to the hands of the Laotians. The study consisted of a before and after comparison of the training's procedural criteria and a longitudinal prospective survey over the last seven months of the programme. The results of the evaluation show that the development of the training conforms to the initial project and that the knowledge acquired in public health methodology by the 12 students is satisfactory. The use of the French language was revealed as being a difficult challenge to overcome for most of the students. Moreover, the partners in Laos had a difficult time playing their role in certain organisational and educational aspects of the training programme. It would be desirable, within the perspective of establishing a Masters of Public Health in Laotian, to establish a partnership between the National Institute of Public Health (INSP) and the Faculty of Medical Sciences (FSM) to work together on the organisation of the training programme. Foreign assistance remains necessary to provide essential financial and educational support. PMID- 15835220 TI - [The gynaecological work by GPs in Brittany]. AB - The aim of this study is to evaluate the gynaecological work done by general practitioners (GPs) in Brittany and the clarify the role that they play in screening for female cancers. Gynaecology represents approximately 10% of the total activity reported by the respondents; however, wide variations have also been noted. This raises important questions on issues such as the competence of these practitioners in this area and equity in access to care and screening, which could potential lead to problems. A large majority of the participating GPs (87%) identified prevention and screening as their primary specific roles to carry out in the care of their female patients. The main reasons for seeking gynaecological consultation are contraception and hormone replacement therapy, and these are the two themes for which GPs request further medical training and continuing education possibilities. Gynaecological work done within the scope of general practice can be hindered by various factors or obstacles: some stem from the doctors themselves (inadequate skills, little interest in this speciality), or rather may be linked to their working conditions (dealing with several reasons for a patient seeking consultation at the same time), while others are a result of how women (or even the doctors themselves) view the tests (reluctance or lack of confidence in the doctors' ability to carry them out properly). In order for GPs to be more involved and implicated in this field, there would need to be the introduction of an efficient and adapted training as well as compatible and effective continuing education programmes suitable to respond to the needs and requirements of the GPs practicing in this area. PMID- 15835221 TI - ["Evaluating health workshops": the experience of health education committees in the Pays de la Loire region]. AB - The five health promotion and education committees (CODES) from the Pays de la Loire region are carrying out health promotion projects involving socially disadvantaged people. These projects are referred to as ateliers sante (literally meaning "health workshops"). These interventions are conducted over a series of participatory sessions, which creates a more supportive structure to guide people with difficulties assisting them to better recognise and formulate their health needs, as they are normally part of a population group which is neither accustomed to expressing their health needs nor to being heard on this subject. At present little is known about these projects; therefore the Inter-regional and departmental health education committee from Nantes chose to carry out an evaluation of 7 of the 47 existing "workshops". Each one of the seven interventions required an average of 31 hours of preparation. Overall, there were 9 sessions organised with 8 to 9 participants, each lasting for the duration of 2.5 hours, held over a period of 32 weeks. At the end of each session, 85% of the participants claimed that there were satisfied. When questioned six months after the end of their "workshop", half of the 37 participants reported that their perceived health status was a little or much better after having taken part in the process. The evaluation demonstrates that a more clear definition of the operational objectives is necessary in order to promote increased input from the participants and to clarify the intentions of the project team towards a population group which needs a means by which it can enter into this critical debate. The process appears to be in accordance with the criteria and goals of health promotion: negotiation of the content, participation of members of the target audience throughout various stages of the project, and adjustment of the size and scale of the project being to involve a small total number of participants in order to favour effective follow up. Although evaluating the effectiveness and impact of such interventions is difficult and complicated by complex methodological questions, these "workshops" seem to have been greatly appreciated both by those who requested them and those who participated in them, with encouraging results having been witnessed in the lives of certain participants, which all serves as an incentive to pursue this kind of project. PMID- 15835222 TI - [Identification of malfunctioning in the treatment and quality of care of maternal morbidity in Abidjan, Cote d'Ivoire]. AB - A study was carried out using a clinical audit aimed at identifying the dysfunctions in the care of female patients with serious morbidity. The study was done at the University Hospital of Cocody (CHU de Codody) and in the health training unit in the southern part of Abobo (Abidjan) from January to May 2000. The study allowed us to track and record the frequency of women who nearly died (40.4%) in both of the sites during the period of the study. Malfunctions were found at all stages of the female patients' care. The provision of medical care during the patients' hospitalisation and care provided in the emergency room were the cases and situations wherein the most frequency was noted, with 42.8% and 39.6% of dysfunctions found respectively. The delay for patients to wait to receive care was long, varying from 80 minutes to 5 days coupled with a lack of follow-up and surveillance of patients. This data demonstrates the inadequacy of the quality of obstetrical care. PMID- 15835223 TI - Evaluation of the quality of venous leg ulcer care given in a multidisciplinary specialist centre. AB - OBJECTIVE: To determine the quality of venous leg ulcer care given in a multidisciplinary, specialist wound-healing centre and to identify problem areas that might affect it. METHOD: The case records of 90 consecutive patients with venous leg ulcers, diagnosed and treated at the Copenhagen Wound Healing Centre, Denmark, were retrospectively audited by a Scandinavian cross-sectional and multidisciplinary expert panel. Quality of care was audited in each case using implicit criteria. The experts then formulated key recommendations for good clinical practice for patients with venous leg ulcers. RESULTS: Quality of care was satisfactory in 74 patients (82%). The one-year healing rate was 77% (69/90), with a three-month recurrence rate of 12% (11/90). Identified problem areas included the lack of systematic assessment of patients' suitability for surgery; lack of systematic, duplex-verified diagnoses of venous aetiology; and the lack of systematic examination of distal arterial pressure. The recommendations include the need for venous diagnosis, differential diagnosis, compression therapy, surgery, systemic treatment, access to venous leg ulcer care and better communication. CONCLUSION: The quality of venous leg ulcer care given in this multidisciplinary centre was satisfactory. PMID- 15835224 TI - A comparison of laser Doppler imaging with other measurement techniques to assess burn depth. AB - Evaluation of burn depth is crucial in supporting treatment decisions, yet none of the commonly used methods offer clinicians the desired level of accuracy. This article reviews the literature to determine the advantages of laser Doppler imaging. PMID- 15835225 TI - Silver absorption and antibacterial efficacy of silver dressings. AB - OBJECTIVE: To evaluate the patterns of silver release from selected sustained silver-release dressings and the protective role of proteins in wound exudate and wound scale. The bactericidal action of silver in chronic wound therapy is also examined. METHOD: Sequential microbiological examination of wound swabs from seven patients with chronic wounds and sampling of wound exudate and wound scale. Silver content was measured using atomic absorption spectrometry. The ability of Contreet Foam to absorb exudate and release silver was studied in punch biopsy wounds in a rodent model. RESULTS: Silver accumulation in wound exudate correlated well with its viscosity and protein content. Silver bound to wound scale and debris was approximately proportional to the silver ion release from dressings. Bacterial burden was controlled, but not eliminated, following chronic silver therapy. CONCLUSION: Silver dressings (Acticoat-7, Actisorb Silver, Contreet Foam, Aquacel Ag and Flamazine) were found to be safe for use in chronic wound therapy. Excess silver ion is bound by wound exudate and wound scale as a protective mechanism. Silver-release dressings are not likely to result in germ free wounds. Further studies are needed to examine potential silver resistance. PMID- 15835226 TI - Estimating costs of pressure area management based on a survey of ulcer care in one Irish hospital. AB - OBJECTIVE: Pressure ulceration remains a significant cause of morbidity for patients and has a real economic impact on the health sector. Studies to date have estimated the cost of management but have not always given a breakdown of how these figures were calculated. There are no published studies that have estimated the cost of management of pressure ulcers in Ireland. A two-part study was therefore undertaken. Part one determined the prevalence of pressure ulcers in a 626-bed Irish acute hospital. Part two set out to derive a best estimate of the cost of managing pressure ulcers in Ireland. METHOD: The European Pressure UlcerAdvisory Panel (EPUAP) minimum data set tool was used to complete the prevalence survey. Tissue viability nurses trained in the data-collection tool collected the data. A cost was obtained for all items of care for the management of one patient with three grade IV pressure ulcers over a five-month period. RESULTS: Of the patients, 2.5% had pressure ulcers. It cost Euros 119,000 to successfully treat one patient. CONCLUSION: We estimate that it costs Euros 250,000,000 per annum to manage pressure ulcers across all care settings in Ireland. PMID- 15835227 TI - Using topical negative pressure therapy to resolve wound failure following perineal resection. AB - Elective resection of the rectum and perineum in a patient with recurrent squamous cell carcinoma of the anus resulted in significant wound failure. Full healing was achieved with topical negative pressure, with no recurrence of the cancer. PMID- 15835228 TI - Does the use of a cleanser on skin surrounding pressure ulcers in older people promote healing? AB - OBJECTIVE: To determine whether the use of a skin cleanser on the skin surrounding pressure ulcers helps to promote healing. METHOD: The study was conducted over a two-year period in patients with stage II or more pressure ulcers. All subjects were at least 65 years of age and resident in a long-term care hospital. During the first year, skin was cleansed with normal saline. In year two, a pH-balanced cleanser was used. The healing times for the two methods were then compared. RESULTS: Healing time was shorter in the group using the cleanser for every stage of ulcer, with an especially significant difference for stage II ulcers (p=0.002). Analysis using the Cox proportional hazards model found a 1.79-fold improvement in the healing rate of stage II ulcers when the surrounding skin was washed with the cleanser. CONCLUSION: Cleaning the surrounding skin with a cleanser rather than normal saline promotes the healing of pressure ulcers. PMID- 15835229 TI - Regeneration of postoperative wounds in surgical patients [corrected]. PMID- 15835230 TI - Skin grafting techniques for soft-tissue coverage of diabetic foot and ankle wounds. AB - Diabetic foot and ankle wounds frequently break down despite off-loading techniques. Skin grafting offers a simple, durable, minimally invasive and cost effective means of wound closure, resulting in a functional and sensate limb. PMID- 15835231 TI - Using cycloidal vibration to heal venous leg ulcers: a cost-analysis based on retrospective data. AB - Using cycloidal vibration to stimulate the circulation to enhance healing can significantly reduce treatment costs. For the patient, the benefits include faster healing times, a better quality of life and a marked reduction in pain. PMID- 15835232 TI - Wound pain: the need for a more understanding approach. AB - Wound pain is hard to measure, and its treatment has suffered from a lack of knowledge on the part of health-care professionals. This paper describes current approaches, the mechanisms of pain and suggests strategies for improved care. PMID- 15835233 TI - The role of pressure-redistributing equipment in the prevention and management of pressure ulcers. AB - In response to a focus on education and research in pressure ulcer prevention, this paper reviews the evolution of pressure-redistributing mattresses and explores the effectiveness of interface pressure measurement in mattress development. PMID- 15835234 TI - COX selectivity--finding the right balance. PMID- 15835235 TI - Retrospective study of bacterial infective arthritis in 31 dogs. AB - OBJECTIVES: To characterise the presenting signs and pathological changes of canine bacterial infective arthritis in 31 dogs, and to document the response to different treatment regimens. Risk factors that may predispose joints to bacterial infective arthritis and influence the success of treatment were also investigated. METHODS: A retrospective review of cases of bacterial infective arthritis that were presented to three university veterinary referral hospitals over a five-year period (January 1997 to January 2002) was performed. RESULTS: The elbow joint (38 per cent) and stifle joint (44 per cent) were most commonly affected. Radiographic changes consistent with pre-existing osteoarthritis were identified in 14 joints, which had no history of previous surgery (articular or periarticular) or penetrating wound. No significant difference (P = 0.78) was identified between the outcome of combined surgical and medical management, and medical management alone. There were trends for poorer outcomes with increased bodyweight of the dog, longer duration of lameness and a higher nucleated cell count of the affected joint fluid at presentation. The overall infection rate for articular surgical procedures at one institution was 1-3 per cent. CLINICAL SIGNIFICANCE: Medical and/or surgical management were usually successful in resolving infection (94 per cent). However, they were frequently unsuccessful in restoring full joint function; this may in part have been due to the nature of the underlying joint PMID- 15835236 TI - Epidemiology of canine urolithiasis in the Czech Republic from 1997 to 2002. AB - OBJECTIVES: To compare data on the epidemiology of canine urolithiasis in the Czech Republic with that from other countries. METHODS: The records from the Centre for Mineralogical Analysis from 1997 to 2002 were reviewed. The data were obtained from mineralogical analysis of 1366 canine uroliths obtained from patients in the Czech Republic. These included 396 females and 629 males. RESULTS: Sixty-eight breeds plus crossbreeds were identified. Eight breeds plus the crossbreeds accounted for 71.3 per cent of all cases. Males were affected more frequently than females (61.4 per cent versus 38.6 per cent). Struvites significantly predominated in females, while in males calcium oxalates, brushites and cystines were the most common stones. Most of the uroliths (48.9 per cent) were 5 mm or less in dimension. By 2001, struvite was the most frequent (38.5 to 44.1 per cent) urolith, followed by calcium oxalate (26.5 to 32.0 per cent). In 2002, calcium oxalate became the most frequent calculus, followed by struvite, mixed calculi and others. CLINICAL SIGNIFICANCE: Comparison of these results with studies by other authors showed that for most of the monitored parameters there was agreement with respect to the proportions of different breeds within the populations of dogs in different geographical areas. PMID- 15835237 TI - Assessment of corticosteroid-induced alkaline phosphatase as a prognostic indicator in canine lymphoma. AB - OBJECTIVES: To examine the incidence of elevated corticosteroid-induced alkaline phosphatase (sALP) in dogs with lymphoma and to determine if sALP is a reliable prognostic indicator in canine lymphoma. METHODS: The medical records of 62 canine lymphoma patients treated with a combination chemotherapy protocol from 1994 to 2003 at the University of Illinois Veterinary Teaching Hospital were examined. Variables assessed with respect to response rate and remission duration included age, bodyweight, sex, breed, World Health Organization stage (I to V), substage (a or b), pretreatment administration of corticosteroid, and serum levels of alkaline phosphatase, sALP and alanine aminotransferase. RESULTS: sALP was not statistically significant with respect to response rate or duration of remission, nor was preinduction glucocorticoid administration. Stage was significant with respect to achieving remission. CLINICAL SIGNIFICANCE: It was found that sALP is not a useful prognostic indicator for response rate and remission duration in dogs with lymphoma. PMID- 15835238 TI - Treating pedal shortening in a dog by metatarsal distraction osteogenesis. AB - A five-month-old, male German shepherd dog was presented with a left hindlimb deformity. Wearing a cast for a prolonged period to treat left metatarsal fractures at eight weeks of age had resulted in physeal closure of the calcaneus, metatarsals and phalanges. Hyperextension of the phalanges and ulceration of the plantar skin proximal to the main tarsal pad were seen. The left foot was 40 mm shorter than the right. At seven months of age, a modified circular external skeletal fixator (CESF) was used to perform distraction osteogenesis of the metatarsal bones for 28 days. The CESF apparatus was removed after eight weeks when there was radiographic consolidation of regenerate bone. At follow-up evaluations four and 12 months after surgery, the phalangeal hyperextension and plantar skin ulceration had resolved. No lameness was detected. PMID- 15835239 TI - Unusual presentation of avulsion of the lateral head of the gastrocnemius muscle in a dog. AB - A five-and-a-half-year-old, male neutered golden retriever was presented with sudden onset, plantigrade stance of the left pelvic limb one month after a traumatic incident. A diagnosis of avulsion of the lateral head of the gastrocnemius muscle was made based on clinical signs and radiographic assessment. Initial wire reduction and fixation were unsuccessful. However, reduction and fixation with monofilament leader line augmented with a trans-hock external fixator resulted in a successful outcome. The dog had a normal gait 18 months after revision surgery. PMID- 15835240 TI - Centrodistal joint lameness in dogs. AB - Lameness attributable to pathology of the centrodistal joint was investigated in five racing greyhounds, five Border collies and two other dogs. Diagnosis was made by the induction of pain from rotationally stressing the medial aspect of the tarsus, and by the exclusion of tarsal fractures and instabilities. Radiographic changes included plantar ligament entheslopathy and centrodistal joint osteophytosis. In many of the cases, the lameness was poorly responsive to rest and non-steroidal anti-inflammatory drugs. One dog was managed conservatively and became sound. In 10 of 11 cases, surgical promotion of centrodistal ankylosis was associated with a return to soundness. PMID- 15835241 TI - Chylothorax and fibrosing pleuritis secondary to thyrotoxic cardiomyopathy. PMID- 15835242 TI - Draft legislation on supply of POMs: how will it affect you? PMID- 15835243 TI - A natural high. PMID- 15835244 TI - Veterinary education in Chile's open economy. PMID- 15835245 TI - Use of the range area in organic egg production systems: effect of climatic factors, flock size, age and artificial cover. AB - (1) To evaluate the effect of climate, flock size, age and artificial cover on the use of range area, a study was conducted in 1994 to 1998, involving 5 farms with a total of 37 flocks of layers kept in commercial, free-range, organic systems. Flocks were visited regularly during the production period to record the number and distribution of hens on the range. (2) On average 9% of the flock used the range area, but with large variations both within and between flocks, and this was partly influenced by climatic factors. Range use was affected by temperature, wind, precipitation, season and age, and there was a tendency for reduced numbers of hens on the range with the time of day and increasing flock size. (3) Eight flocks (including houses and range areas) were divided into 8 paired units, and the range area of one unit in each of the flocks was provided with artificial cover. There were more hens on the range when artificial cover was present and the presence of cover attracted more hens away from the area immediately outside the hen house. PMID- 15835246 TI - Design of laying nests in furnished cages: influence of nesting material, nest box position and seclusion. AB - (1) Preferences for three nesting materials and nest box positions were investigated simultaneously in two trials using a furnished cage: one with 18 individual laying hens and one with 18 groups of 5 hens. Following a habituation period in pre-test cages, every hen or group of hens was tested for 2 d: once without and once with plastic flaps at the entrance of the nest boxes. (2) Hens preferred peat and artificial turf to coated wire mesh for egg laying. (3) One nest box position was clearly preferred to both other nest boxes. The hens' choice of nest box position was influenced by the pre-test cage in which they had been habituated. (4) The presence of plastic flaps at the entrance of the nest boxes had no influence on the proportion of eggs laid on the different nesting materials or on the proportion of floor eggs. (5) Individual and group testing resulted in the same overall results despite the presence of a distinct group effect. PMID- 15835247 TI - Effects of different management factors on broiler performance and incidence of tibial dyschondroplasia. AB - (1) This study was to determine the effects of two lighting programmes (continuous--24L:0D or intermittent--12L:3(1L:3D)), two feeding programmes (continuous--ad libitum or meal Jeeding--3 h feed:3 h deprivation x 4 times per d) and two ascorbic acid (AA) supplementations (0 or 150 mg/l in water) on broiler performance and incidence of tibial dyschondroplasia (TD). (2) One-day old male commercial (Ross PM3) broiler chicks (1600) were used. The chicks were randomly divided into 8 treatment groups (2 x 2 x 2) and their replicates (2) replicates per treatment). (3) Intermittent lighting increased body weight and improved feed conversion. Body weight of broilers was decreased by meal feeding. AA supplementation had no significant effect on the body weight and feed conversion ratio. There was no lighting x feeding x supplemental AA interaction for body weight, feed conversion ratio, mortality rate and carcase weight. (4) Supplemental AA improved the cortical thickness of tibiotarsus and decreased the incidence of TD. (5) It was concluded that the application of intermittent lighting in combination with supplementation of 150 mg/l AA is a useful method to reduce TD in broilers without loss of performance. PMID- 15835248 TI - Rearing greater rhea (Rhea americana) chicks: is adoption more effective than the artificial intensive system? AB - (1) Survival and weight gain of farmed Greater Rhea (Rhea americana) chicks reared by the adult males that adopted them were compared with those of chicks reared under an artificial intensive system. (2) Both variables were periodically recorded up to the age of 3 months. Gompertz growth curves were fitted to individual growth data using the average adult weight of this population as asymptote. (3) No significant differences in survival rate were detected between systems (adoption=47%, intensive=43%). However, during the first half of the breeding season (mid-spring to mid-summer), the growth rate of adopted chicks (0.01481) was higher than that of intensively reared chicks (0.01296). (4) The adoption system may be more effective in terms of growth, and is probably more efficient in cost/effectiveness than the artificial intensive technique most frequently used. Adoption by males has additional advantages, such as a correct imprinting of the chicks and the selection of more capable individuals. Therefore, it should be used not only commercially but also in conservation projects where individuals are released to the wild. PMID- 15835249 TI - Influence of Dermanyssus gallinae and Ascaridia galli infections on behaviour and health of laying hens (Gallus gallus domesticus). AB - (1) The effect of infections with Dermanyssus gallinae (poultry red mite or chicken mite) and Ascaridia galli (roundworm) on the behaviour and health of laying hens was investigated. (2) Six groups of 15 pullets (Isa Brown) were kept in indoor pens from 18 weeks of age. Two groups were artificially infected with D. gallinae, two groups with A. galli and two groups were kept as uninfected controls. The hens were observed for behavioural reactions and physiological changes (weight gain and various blood variables) to the parasitic infections. (3) Infections with D. gallinae resulted in reduced weight gain, anaemia and even death of some of the hens. Behavioural changes were also observed, as the mite infected hens showed higher self-grooming and head scratching both during the day and night. (4) A. galli resulted in a lower weight gain but no significant changes were seen in blood variables or behavioural activities. PMID- 15835250 TI - Genetic parameters of ascites-related traits in broilers: effect of cold and normal temperature conditions. AB - (1) Ascites syndrome is a growth-related disorder of broilers that occurs more often in fast-growing birds and at low temperatures. The objective of this study was to estimate genetic and phenotypic correlations among ascites-related traits measured either under cold or under normal temperature conditions, and to estimate genetic correlations between ascites-related traits measured under cold and normal conditions. (2) Several traits related to ascites were measured on more than 4000 chickens under cold conditions and on more than 700 chickens under normal conditions. (3) The heritability estimates for body weight (BW) measured under cold and normal conditions were 0.42 and 0.50, respectively, for haematocrit value 0.46 and 0.17, respectively, and for ratio of right to total ventricular weight 0.45 and 0.12, respectively. (4) The genetic correlation between BW and haematocrit value under cold conditions was -0.23 and between BW and ratio of right to total ventricular weight -0.27. Under normal conditions, however, these genetic correlations were 0.55 and 0.50, respectively. (5) These results demonstrate that the heritability estimates of ascites-related traits as well as genetic correlations between ascites-related traits and BW depend on the temperature conditions under which animals are kept. (6) Strong positive genetic correlations (around 0.8) were observed between total mortality, fluid in the abdomen and ratio of right to total ventricular weight under cold conditions. The genetic correlation between ratio of right to total ventricular weight under cold and normal conditions was 0.91. (7) These results suggest that the ratio of right to total ventricular weight measured under normal temperature conditions might serve as a good indicator trait for ascites. PMID- 15835251 TI - Genetic parameters of ascites-related traits in broilers: correlations with feed efficiency and carcase traits. AB - (1) Pulmonary hypertension syndrome followed by ascites is a metabolic disorder in broilers that occurs more often in fast-growing birds and at cool temperatures. (2) Knowledge of the genetic relationships among ascites-related traits and performance traits like carcase traits or feed efficiency traits is required to design breeding programmes that aim to improve the degree of resistance to ascites syndrome as well as production traits. The objective of this study was to estimate these genetic correlations. (3) Three different experiments were set up to measure ascites-related traits (4202 birds), feed efficiency traits (2166 birds) and carcase traits (2036 birds). The birds in different experiments originated from the same group of parents, which enabled the estimation of genetic correlations among different traits. (4) The genetic correlation of body weight (BW) measured under normal conditions and in the carcase experiment with the ascites indicator trait of right ventricle to total ventricle ratio (RV:TV) measured under cold conditions was 0.30. The estimated genetic correlation indicated that single-trait selecting for BW leads to an increase in occurrence of the ascites syndrome but that there are realistic opportunities of multi-trait selection of birds for improved BW and resistance to ascites. (5) Weak but positive genetic relationships were found between feed efficiency and ascites-related traits suggesting that more efficient birds tend to be slightly more susceptible to ascites. (6) The relatively low genetic correlation between BW measured in the carcase or in the feed efficiency experiments and BW measured in the ascites experiment (0.49) showed considerable genotype by environment interaction. (7) These results indicate that birds with high genetic potential for growth rate under normal temperature conditions have lower growth rate under cold-stress conditions due to ascites. PMID- 15835252 TI - Heritability of sudden death syndrome and its associated correlations to ascites and body weight in broilers. AB - (1) Genetic parameters for the sudden death syndrome (SDS) were estimated in meat type chickens. Data were collected over 11 generations of selection for body weight within two distinct breeds (Cornish and White Rock). (2) The animal model was used exclusively with linear methods (LM) to estimate genetic parameters. Heritability (h2) of SDS on the liability scale was 0.30 +/- 0.002 and 0.25 +/- 0.002 in the Cornish and White Rock breeds, respectively. (3) A positive genetic correlation (r(g)) with ascites (AS) was determined (approximately 0.3 +/- 0.006). However, it was not possible to estimate the rg of SDS with body weight because of the low prevalence of the defect trait studied (1.8% in the Cornish and 1-5% in the White Rock). (4) Heritability of SDS calculated using male records only was 0.45 +/- 0.009 and 0.35 +/- 0.009, and r(g) with body weight was 0.30 +/- 0.010 and 0.27 +/- 0.009, in the Cornish and White Rock breeds, respectively. (5) In conclusion, the heart defect investigated was heritable with a positive genetic correlation with AS and body weight. PMID- 15835253 TI - Use of Trichosporon mycotoxinivorans to suppress the effects of ochratoxicosis on the immune system of broiler chicks. AB - (1) The objective of this study was to determine whether the dietary inclusion of Trichosporon mycotoxinivorans (TRM) could suppress the detrimental effects of ochratoxin A (OTA) on the immune system of broiler chicks. (2) Six experimental treatments were tested in 300 1-d-old broiler chicks. Treatments included addition to a standard broiler ration of neither OTA nor TRM (Diet 1), OTA alone (500 microg/kg), OTA plus TRM at three inclusion rates (10(4) CFU/g of feed, 10(5) CFU/g, 10(6) CFU/g) and TRM alone at 10(5) CFU/g of feed. The ration was fed to chicks for 42 d. (3) Blood samples were collected at d 10, 20, 30 and 40 and macrophages and heterophils were isolated. The following variables were determined in macrophages and heterophils activated by phorbol myristate acetate (65 microM): cell viability, total cell-associated urokinase-plasminogen activator (u-PA), membrane-bound u-PA, free u-PA binding sites and superoxide production. (4) There was a decrease in the viability of macrophages and heterophils from chicks receiving OTA-contaminated feed compared to the viability of cells from control birds at d 40. Dietary TRM completely blocked the effect of OTA on cell viability; all three inclusion rates were equally effective. There was a decrease in total cell-associated and membrane-bound u-PA in macrophages and heterophils of chicks receiving OTA-contaminated feed compared to the corresponding values in control birds for heterophils at d 30 and 40 and for the macrophages at d 40. (5) Similarly, dietary TRM abolished the effect of OTA on total cell-associated and membrane-bound u-PA activity. All three inclusion rates of yeast were equally effective. Heterophils, but not macrophages, isolated from chicks receiving OTA-contaminated diet produced less superoxide anion compared to all other diet groups at d 30 and 40. (6) The immune system is a primary target of OTA toxicity in broilers: several functional properties of macrophages and heterophils were depressed in chicks fed OTA-contaminated feed. There was a delay of 30d before the immunosuppressive effect became apparent. The dietary inclusion of TRM completely blocked the detrimental effects of OTA on several immune properties in broilers. PMID- 15835254 TI - Soft wheat instead of hard wheat in pelleted diets results in high starch digestibility in broiler chickens. AB - (1) The aim of the experiment was to re-examine variations in digestibilities of food components in 3-week-old broiler chickens fed on pelleted diets based on wheats differing in lipase activity and hardness. Fourteen wheat (Triticum aestivum) samples, each from a different cultivar, were included at 550 g/kg in 14 different diets given to male Ross broiler chicks from 7 d of age. The other main ingredients consisted of soyabean meal (353 g/kg) and rapeseed oil (55 g/kg). A 15th diet containing durum wheat (Triticum durum) was also tested. (2) Hardness of wheats (Triticum aestivum) varied between 14 (very soft) and 88 (very hard), and lipase activity of wheats varied from 1 to 13.1 (relative scale). No significant correlation was found between in vitro viscosities and other parameters such as hardness, particle size of wheat flours and lipase. Hardness was correlated with the mean particle size of wheat flours and durability of pellets. (3) Individual lipid digestibilities were negatively correlated with in vitro viscosities of wheats. (4) Individual starch digestibilities were negatively correlated with wheat hardness, particle size of wheat flour before pelleting, and pellet durability. The ratio of measured AME(N) to predicted AME(N) was also negatively correlated with wheat hardness. Simple regression calculation showed that a 100-point increase in wheat hardness resulted in a 3% decrease in the AME(N) value of diets. Multiple regression calculation showed the food/gain ratio (d 10 to d 21) to be positively related to wheat hardness and negatively related to pellet durability. (5) Wheat lipase activity was positively correlated with individual starch digestibility, which was the reverse of a result obtained in a previous experiment. Thus, wheat lipase activity did not seem consistent for predicting starch digestibility and AME(N) values. (6) Among all wheat samples, durum wheat showed the highest protein content and the lowest content of water-insoluble cell-wall. Starch digestibility of durum wheat tended to be lower than that of other wheats (0.916 vs 0.936). However, no significant difference in AME(N) was observed between the durum wheat sample and other wheats. (7) Gut morphometric data measured at d 24 did not show significant differences between dietary treatments. PMID- 15835255 TI - Effect of enzyme supplementation on the metabolisable energy content of solvent extracted rapeseed and sunflower seed meals for chicken, guinea fowl and quail. AB - (1) The nitrogen-corrected apparent metabolisable energy (AME(N)) content of solvent-extracted rapeseed and sunflower seed (un-decorticated) meals in relation to species (chicken, guinea fowl and quail) and dietary addition of feed enzymes (0 or 0.5 g/kg diet) was evaluated by a diet replacement method in a 3 x 2 factorial design. (2) The metabolism trial was conducted at two substitution levels (200 and 400 g/kg diet) of each meal with or without supplementation of commercial enzyme preparation in 6 individuals or 6 groups of cockerels, guinea fowls and quails. (3) The experimental diets were fed for a period of 12 d followed by a 3-d collection period during which total feed consumed and droppings output were quantitatively recorded. (4) The AME(N) values of rapeseed meal for cockerels, guinea fowls and quails were 8.4, 8.7 and 8.8 MJ/kg, respectively, while the corresponding values for sunflower seed meal were 6.1, 6.1 and 6.2 MJ/kg dry matter, without enzyme supplementation. (5) The AME(N) value of rapeseed meal did not improve with enzyme supplementation. However, AME(N) values of sunflower seed meal significantly increased with enzyme supplementation, from 6.1 to 6.5 MJ/kg dry matter. (6) Since AME(N) values of rapeseed meal and sunflower seed meal were similar in chicken, guinea fowl and quail, values reported for chicken could, therefore, be used for guinea fowl and Japanese quail. PMID- 15835256 TI - Effects of dietary conjugated linoleic acid and high-oleic sunflower oil on performance and egg quality in laying hens. AB - (1) Laying hen performance, yolk fatty acid (FA) concentrations, sensory quality and firmness of eggs were evaluated with respect to the inclusion in the diet of conjugated linoleic acid (CLA) and high-oleic acid sunflower oil (HOSO). (2) Nine diets were arranged factorially, with three concentrations of CIA (0, 1 and 2 g/kg) and HOSO (10, 20 and 30 g/kg). (3) Type of diet did not affect egg production traits. (4) Dietary addition of CLA decreased yolk lipid content and yolk lipid concentrations of monounsaturated FA, C(20:4 n-6) and C(22:6 n-3), but increased those of CLA and saturated FA. (5) Dietary addition of HOSO increased monounsaturated FA concentrations in the yolk lipid but decreased those of CLA and saturated FA. (6) CLA supplementation increased yolk moisture and firmness and impaired the sensory quality of eggs. (7) An interaction between CLA and HOSO addition was found as effects of CLA addition on yolk lipid CLA concentrations and egg quality traits were smaller when the amount of HOSO in the diet increased. (8) Regression equations have been calculated in order to predict yolk CLA and C(18:1), concentration from dietary composition, and yolk firmness from yolk FA composition. PMID- 15835257 TI - Effects of dietary boron supplementation on some biochemical parameters, peripheral blood lymphocytes, splenic plasma cells and bone characteristics of broiler chicks given diets with adequate or inadequate cholecalciferol (vitamin D3) content. AB - (1) The effects of 5 and 25 mg/kg boron supplementation of diets with inadequate (6.25 microg/kg) or adequate (50 microg/kg) cholecalciferol (vitamin D3) content on some biochemical parameters, tibia characteristics, peripheral blood lymphocyte and splenic plasma cell counts of broilers were investigated. (2) Supplementation of the diet with boron affected plasma concentrations of boron, iron, copper and zinc and also tibia boron, zinc and calcium concentrations but did not have any effect on tibia iron or copper concentrations or tibia ash and tibia weight values. (3) Boron supplementation caused significant increases in splenic plasma cell count but decreased the proximal and distal tibia growth plate widths. There was no effect of boron supplementation on peripheral blood alpha-naphthyl acetate esterase (ANAE) content. Whole blood haematocrit and haemoglobin counts were significantly increased by boron supplementation but there were no effects on leucocyte ratios such as eosinophil, basophil, monocyte, lymphocyte and thrombocyte. (4) In general, the findings of the present study support the hypothesis that boron has an important biological role that affects the mineral metabolism of animals by influencing both biochemical and haematological mechanisms. PMID- 15835258 TI - Breeder hen dietary L-carnitine affects progeny carcase traits. AB - (1) Ross 308 broiler breeder hens were given diets containing 0 or 25 mg L carnitine/kg from 21 weeks of age. (2) Hens were inseminated with semen from Ross broiler breeder males and subsequent growth performance and carcase traits, of progeny obtained from hatches at 30, 35 and 37 weeks of age, were evaluated. (3) Progeny were hatched in a common facility and separated by gender. Experimental treatments employed for the 30-, 35- and 37-week hatches, respectively, were: hen diet and progeny gender (16 replications with two subplots); hen diet, progeny diet (0 and 50 mg L-carnitine/kg of diet) and progeny gender (16 replications with 4 subplots); and hen diet and progeny diet (high and low density; 16 replications with two subplots). (4) Females had lower growth rate and less breast meat, but greater proportions of carcase fat and breast meat than males. Growth performance measurements of progeny were not affected by hen L-carnitine, but hen L-carnitine decreased abdominal fat in progeny. Increasing diet density in the chick diets increased growth and carcase weights. Hen and progeny dietary L-carnitine interacted to increase male mortality. However, dietary hen L carnitine decreased carcase fat and increased breast meat in progeny fed on high nutrient density diets. (5) In conclusion, L-carnitine in the diet of hens affected carcase traits of their progeny. PMID- 15835259 TI - Dietary n-3 and n-6 fatty acids alter avian glucose metabolism. AB - (1) This investigation studied the effects of dietary saturated and polyunsaturated fatty acids (PUFAs) from the n-3 and n-6 series on insulin action and glucose uptake in broiler chickens. (2) One-day-old male chicks were fed on a commercial starter diet for 3 weeks, randomly divided into three groups (n = 6) and fed ad libitum on isonitrogenous experimental diets of equal energy density for a further 6 weeks. The diets contained 20.8 g/100 g protein and 80 g/kg of either edible tallow, fish oil or sunflower oil, giving diets high in saturated fatty acids, n-S PUFAs or n-6 PUFAs, respectively. (3) Jugular catheterisation was performed under general anaesthesia during week 4 of the dietary treatments and the birds given 7 d post-surgery to recover. To estimate insulin action, a bolus glucose infusion (1 g/kg) was given to each chicken and sequential blood samples taken over a one-hour period. To estimate the disappearance rate of glucose from the plasma and its incorporation into tissues, 2-deoxy-D-3H glucose (2DG-3H glucose) was infused into each chicken (50 microCi) 2 d later. (4) Although there were no significant differences in glucose clearance rate following the glucose infusion, the maximal insulin release in response to the glucose infusion was higher in the tallow group than in either the n-3 or n-6 PUFA dietary groups. There were no significant differences in the clearance rate of 2DG-3H glucose. Labelled glucose incorporation into the breast muscle was greater in birds given fish oil than in birds given tallow and significantly greater than in birds given sunflower oil. (5) The data suggest that the type of dietary fat can influence glucose metabolism and that this change in glucose utilisation may alter the energy metabolism of the broiler. PMID- 15835260 TI - Sustained availability of trimethoprim in drinking water to achieve higher plasma sulphonamide-trimethoprim antibacterial activity in broilers. AB - (1) In order to make trimethoprim (TMP) available to broilers throughout the day, a sustained release formulation (SRF) of the drug in the form of granules was added to the water tank that supplies drinking water. (2) Broilers were initially dosed with sulphachloropiridazine-TMP (SCP-TMP 5:1) and then further medicated throughout the day, achieving in the end a dose of 30 mg/kg each of SCP and TMP (group A). Group B received a preparation with the same dose of SCP and TMP (1:1) as group A, but administered as a single dose without the SRF of TMP. Group C received the customary SCP-TMP 5:1 preparation (30 and 6 mg/kg, respectively). Water tanks were completely consumed in 3 to 4 h. (3) Broilers were bled at different times and concentration of antibacterial activity in serum determined by correlating the composite antibacterial activity of SCP and TMP with actual concentrations of these drugs by means of a microbiological agar diffusion assay. (4) Time vs serum concentrations of activity were higher in group B; the increments in the maximum serum concentration for group B over groups A and C being 39 and 67%, respectively. (5) However, the sustained concentration of activity over time, measured as the area under the cu)rve, was highest in group A. Group B had higher values for area under the curve than group C. (6) An additional dose of TMP to achieve 30 mg/kg of both SCP and TMP improves the serum concentration of this combination over the customary 5:1 proportion. The best values for sustaining antibacterial activity were obtained using a 1:1 ratio as in group A. The use of a SRF as in group A may translate into better clinical results. PMID- 15835261 TI - Fertility and hatchability in RIR and WL breeds as functionally modified by crossing them in alternate sex combinations (Gallus domesticus). AB - (1) Four breeding groups of Rhode Island Red and White Leghorn domestic fowl (RIR (female) x RIR (male), RIR (female) x WL (male), WL (female) x RIR (male) and WL (female) x WL (male)) were compared for fertility, hatchability, and their post insemination sustainability, egg weight loss during incubation and uncovered yolk in abdominal cavity of dead in shell in order to understand the problems associated with the RIR breed in these respects. (2) Crossing RIR (female) with WL (male) or in reverse sex combinations did not improve fertility in comparison to pure RIR chickens and all these groups were less fertile than the pure WL. (3) Unlike fertility, hatchability in RIR improved with the change to either sex partner of the WL breed but the WL (female) x RIR (male) combination was similar to the pure WL (97.72 and 97.12%, respectively). In contrast, crossing RIR (female) with WL (male) resulted in an improvement (86.67%) as compared to pure RIR (76.67%) but still lower than the pure WL and WL (female) x RIR (male) cross. (4) Egg weight loss during incubation was more (20.16%) in pure RIR as compared to RIR (female) x WL (male) (17.13%), followed by WL (female) x RIR (male) (10.28%) and pure WL (9.57%). (5) There were more dead-in-shell embryos with yolks outside their abdominal cavity in pure RIR and their crosses as compared to pure WL breeds. (6) Fertility was sustained for longer in WL than other combinations with post-artificial insemination using constant number of spermatozoa. Fertility after a week of insemination tended to decrease more rapidly than hatchability on a fertile egg basis. (7) It is concluded that both sexes are responsible for the poor fertility in RIR but the female is responsible for poor hatchability and this poor performance is mainly due to greater egg weight loss during incubation. PMID- 15835262 TI - The role of nerve growth factor receptors in cholinergic basal forebrain degeneration in prodromal Alzheimer disease. AB - Dysfunction of nerve growth factor (NGF) and its high (TrkA) and low (p75NTR) affinity receptors has been suggested to underlie the selective degeneration of the nucleus basalis (NB) cholinergic cortical projection neurons in end stage Alzheimer disease (AD). Whether the NGF system is dysfunctional during the prodromal stages of AD has only recently been evaluated. Surprisingly, the number of choline acetyltransferase-containing neurons remains stable despite a significant reduction in NGF receptor-positive cells in people with mild cognitive impairment (MCI), suggesting a phenotypic NGF receptor downregulation but not a frank loss of NB neurons during prodromal AD. Moreover, there is a loss of cortical TrkA in the face of stable p75NTR and increased proNGF levels, the precursor molecule of mature NGF, in early AD. Depending upon the cellular context these changes may result in increased pro-apoptotic signaling, cell survival, or a defect in retrograde transport mechanisms. Alterations in NGF and its receptors within the cholinotrophic NB system in early AD suggest that NGF mediated cell signaling is required for the longterm survival of these neurons. Therapeutic neurotrophic intervention might delay or prevent NB neuron degeneration and preserve cholinergic cortical function during prodromal AD. PMID- 15835263 TI - Accreditation council for graduate medical education (ACGME) competencies in neuropathology training. AB - The Accreditation Council for Graduate Medical Education (ACGME) has defined 6 core competencies for all physicians: patient care; medical knowledge; practice based learning and improvement; interpersonal and communication skills; professionalism; and systems-based practice. However, the specific wording of the descriptions often assumes that the physician is a clinician rather than a pathologist. Therefore, the American Association of Neuropathologists, Inc. asked its Professional Affairs Committee to examine the core competencies and determine how they relate to training in neuropathology. The Committee's report is presented here in 6 sections, corresponding to the 6 competencies. In each section, the ACGME definition of that particular competency is either quoted directly or, more often, modified slightly to clarify how the competency applies to neuropathology. Each of the defined competencies is then followed by possible assessment tools, selected from those recommended in the ACGME's "toolbox." Specific suggestions are given for designing tools that apply to neuropathology. Many of the suggested activities and documentation methods can be combined into efficient, carefully formulated training/evaluation exercises. Different tools may be more applicable in some training programs. PMID- 15835264 TI - Hereditary ferritinopathy: a novel mutation, its cellular pathology, and pathogenetic insights. AB - We report a family of French Canadian and Dutch ancestry with hereditary ferritinopathy (neuroferritinopathy) and a novel mutation (C insertion at nt646 647 in exon 4) in the ferritin light chain gene, resulting in a longer than normal protein. Our failure to immunostain most of the abnormal ferritin deposits in the proband with a conformation-dependent monoclonal antibody to ferritin light chain supported a previously postulated conformational change of ferritin light chain in this disease. The posterior putamen and cerebellum were the primary pathologic loci in our proband, but asymptomatic hepatocytic intranuclear accumulations of iron and ferritin also were present. Both neurons and glia displayed highly distinctive, if not pathognomonic, swollen to vacuolated nuclei containing ferritin and iron. Hyaline deposits, again staining for both ferritin and iron, were additional morphologic features that may be unique to the ferritinopathies. The iron, at least in putamen where there was a nearly 40-fold increase, appeared to be both in the ferrous (Fe2+) and ferric (Fe3+) form; it was the most likely cause of the observed neuronal and glial apoptosis. We found morphologic evidence of both lipid peroxidation and abnormal nitration of proteins in putaminal neurons and glia, confirming the expected oxidative stress due to this excessive iron. Biochemical and immunohistochemical abnormalities in mitochondria also were demonstrated, probably due to an imbalance in iron homeostasis that had a deleterious effect on the respiratory chain. PMID- 15835265 TI - Progressive loss of motor neuron function in wasted mice: effects of a spontaneous null mutation in the gene for the eEF1 A2 translation factor. AB - Wasted (wst) is a spontaneous autosomal recessive mutation in which the gene encoding translation factor eEF1A2 is deleted. Homozygous mice show tremors and disturbances of gait shortly after weaning, followed by motor neuron degeneration, paralysis, and death by about 28 days. We have now conducted a more detailed analysis of neuromuscular pathology in these animals. Reactive gliosis was observed at 19 days postnatal in wst/wst cervical spinal cord, showing a rostrocaudal gradient. This was followed a few days later by motor neuron vacuolation and neurofilament accumulation, again with a rostrocaudal progression. Thoracic/abdominal muscles from wst/wst mice aged 17 days showed evidence of progressive denervation of motor endplates, including weak synaptic transmission and retraction of motor nerve terminals. Similar abnormalities appeared in distal, lumbrical muscles from about 25 days of age. We conclude that spontaneous failure of eEF1A2 expression in the wasted mutant first triggers gliosis in spinal cord and retraction of motor nerve terminals in muscle, and then motor neuron pathology and death. The early initiation and rapid progression of motor unit degeneration in wst/wst mice suggest that they should be considered an important and accessible model of early-onset motor neuron degeneration in humans. PMID- 15835266 TI - The FcRgamma chain is not essential for induction of experimental allergic encephalomyelitis (EAE) or anti-myelin antibody-mediated exacerbation of EAE. AB - Macrophages are considered essential mediators in multiple sclerosis (MS) pathogenesis, presumably through myelin phagocytosis and release of inflammatory mediators. Macrophages and microglia express activating Fcgamma receptors (FcgammaRI and FcgammaRIII), which depend on the FcRgamma chain for surface expression and signaling. In MS lesions, crosslinking of FcgammaR by immunoglobulins (IgG) directed against myelin may enhance myelin phagocytosis and inflammation. We studied the role of FcgammaR and anti-myelin antibodies in MOG35 55-induced experimental allergic encephalomyelitis (EAE) in C57BL/6 mice, a model of MS-like disease. Incidence and severity of EAE were similar in FcRy chain-/- (FcRgamma-/-) and wild-type (wt) mice, albeit with delayed onset in FcRgamma-/- mice. This demonstrates that the FcRy chain is not essential for induction of EAE, but that FcRgamma signaling may contribute to the preclinical phase. The role of FcgammaR in antibody-mediated demyelination was addressed by injection of anti-myelin antibodies (Z12 mAb) at onset of MOG35-55-induced EAE. Injection of Z12 mAb rapidly reduced survival time in both wt and FcRgamma-/- mice, demonstrating that antibody-mediated exacerbation of EAE is independent of the FcRgamma chain. Interestingly, Z12-induced exacerbation of inflammation and demyelination persisted longer in wt than FcRgamma-/- mice, suggesting that IgG FcgammaR interactions may contribute to a sustained pathologic effect of anti myelin antibodies in the CNS. PMID- 15835267 TI - Chromosomal instability in meningiomas. AB - Approximately 60% of sporadic meningiomas are caused by inactivation of the NF2 tumor suppressor gene on chromosome 22. No causative gene is known for the remaining 40%. Cytogenetic analysis shows that meningiomas caused by inactivation of the NF2 gene can be divided into tumors that show monosomy 22 as the sole abnormality and tumors with a more complex karyotype. Meningiomas not caused by the NF2 gene usually have a diploid karyotype. Here we report that, besides the clonal chromosomal aberrations, the chromosome numbers in many meningiomas varied from one metaphase spread to the other, a feature that is indicative of chromosomal instability. Unexpectedly and regardless of genotype, a subgroup of tumors was observed with an average number of 44.9 chromosomes and little variation in the number of chromosomes per metaphase spread. In addition, a second subgroup was recognized with a hyperdiploid number of chromosomes (average 48.5) and considerable variation in numbers per metaphase. However, this numerical instability resulted in a clonal karyotype with chromosomal gains and losses in addition to loss of chromosome 22 only in meningiomas caused by inactivation of the NF2 gene. In cultured cells of all tumor groups, bi- and multinucleated cells were seen, as well as anaphase bridges, residual chromatid strings, multiple spindle poles, and unseparated chromatids, suggesting defects in the mitotic apparatus or kinetochore. Thus, we conclude that even a benign and slow-growing tumor like a meningioma displays chromosomal instability. PMID- 15835269 TI - Dysferlin interacts with affixin (beta-parvin) at the sarcolemma. AB - The dysferlin gene is defective in Miyoshi myopathy (MM) and limb girdle muscular dystrophy type 2B (LGMD2B). Dysferlin is a sarcolemmal protein that is implicated in calcium-dependent membrane repair. Affixin (beta-parvin) is a novel, integrin linked kinase-binding protein that is involved in the linkage between integrin and the cytoskeleton. Here we show that affixin is a dysferlin binding protein that colocalizes with dysferlin at the sarcolemma of normal human skeletal muscle. The immunoreactivity of affixin was reduced in sarcolemma of MM and LGMD2B muscles, although the total amount of the affixin protein was normal. Altered immunoreactivity of affixin was also observed in other muscle diseases including LGMD1C, where both affixin and dysferlin showed quite similar changes with a reduction of sarcolemmal staining with or without cytoplasmic accumulations. Colocalization of dysferlin and affixin was confirmed by immunofluorescence analysis using dysferlin-expressing C2 myoblasts. Wild-type and mutant dysferlin colocalized with endogenous affixin. The interaction of dysferlin and affixin was confirmed by immunoprecipitation study using normal human and mouse skeletal muscles. Using immunoprecipitation with deletion mutants of dysferlin, we have identified that C-terminal region of dysferlin is an apparent binding site for affixin. We also found N-terminal calponin homology domain of affixin as a binding site for dysferlin. Our results suggest that affixin may participate in membrane repair with dysferlin. PMID- 15835268 TI - Molecular, anatomical, and biochemical events associated with neurodegeneration in mice with Niemann-Pick type C disease. AB - In Niemann-Pick type C (NPC) disease, cholesterol associated with either apoE or apoB100 is taken up by cells in all tissues, including the central nervous system, through clathrin-coated pits and becomes trapped in late endosomes and lysosomes. This study defines the functional, biochemical, and molecular events that ensue as nerve cell death occurs. In mice homozygous for a mutation in NPC1, neuromuscular dysfunction begins at 5 weeks and death occurs at 13 weeks of age. Cholesterol accumulates in every tissue in the body. Purkinje cell loss in the cerebellum begins at 3 to 4 weeks of age and is nearly complete by 11 weeks. This neurodegeneration in the cerebellum is associated with increases in the levels of mRNA for caspase 1, caspase 3, NPC2, LipA, apoE, apoD, glial fibrillary acidic protein, and tumor necrosis factor-alpha, but not for most target genes of the LXR nuclear receptors. The level for apoER2 is significantly reduced. These studies show there is a compensatory increase in NPC2 and LipA in an attempt to overcome the physiological defect caused by the mutation. Nevertheless, neurodegeneration proceeds utilizing apoptosis with activation of glial cells, increased apoE and apoD synthesis, and increased cholesterol turnover across the CNS. PMID- 15835270 TI - Lhermitte-Duclos disease: a report of 31 cases with immunohistochemical analysis of the PTEN/AKT/mTOR pathway. AB - Lhermitte-Duclos disease (LDD) is a rare cerebellar tumor associated with Cowden disease (CD) and germline mutations in the PTEN gene. To further define these relationships, we reviewed clinical and pathologic findings in 31 LDD cases and analyzed the status of the PTEN pathway in 11 of them. We hypothesized that the granule cell hypertrophy in LDD is secondary to activation of mammalian target of rapamycin (mTOR), a downstream effector in the PTEN/AKT pathway and a major regulator of cell growth. Histopathologically, in addition to the classical findings of LDD, we observed prominent vascular proliferation and vacuolization of the white matter in many of the lesions. Four patients met diagnostic criteria for CD, and many of the remaining patients had some clinical features of CD. Immunohistochemical analysis showed high levels of phospho-AKT and phospho-S6 in the large ganglionic cells forming the lesions, indicating activation of the PTEN/AKT/mTOR pathway and suggesting a central role for mTOR in the pathogenesis of LDD. These data support recommendations for genetic testing and screening for CD in patients with LDD and suggest a novel therapy for LDD through pharmacologic inhibition of mTOR. PMID- 15835272 TI - JCAHO, public are watching: act now to improve cardiac core measure data. PMID- 15835271 TI - The syntrophin-dystrobrevin subcomplex in human neuromuscular disorders. AB - The syntrophins and alpha-dystrobrevin form a subcomplex with dystrophin at the skeletal muscle membrane, and are also highly concentrated at the neuromuscular synapse. Here we demonstrate that the different syntrophins and alpha dystrobrevin isoforms have distinct expression patterns during human skeletal muscle development, and are differentially affected by loss of dystrophin anchorage and denervation in human neuromuscular disease. During normal fetal development, and in Duchenne muscular dystrophy and denervation disorders, alpha1 syntrophin and alpha-dystrobrevin are absent or markedly reduced at the sarcolemmal membrane. beta1-Syntrophin is the predominant syntrophin isoform expressed at the muscle membrane during development, and it undergoes upregulation in response to loss of alpha1-syntrophin in Duchenne muscular dystrophy and in denervation. Upregulation of beta1-syntrophin in neuromuscular disorders is associated with re-expression of the fetal nicotinic acetylcholine receptor gamma-subunit, cardiac actin, and neonatal myosin, suggesting reversion of muscle fibers to an immature phenotype. We show that denervation specifically affects expression of the syntrophin-dystrobrevin subcomplex and does not affect levels or localization of other members of the dystrophin-associated protein complex. Our results confirm that dystrophin is required for anchorage of the syntrophin-dystrobrevin subcomplex and suggest that expression of the syntrophin dystrobrevin complex may be independently regulated through neuromuscular transmission. PMID- 15835273 TI - AHA program can have big impact on your data. PMID- 15835274 TI - Discharge planning advisor. New system streamlines patient throughput. PMID- 15835275 TI - Discharge planning advisor. Is it redundant to have DPs in addition to CMs? PMID- 15835276 TI - ICU reduces mortality of critically ill ICU patients. PMID- 15835277 TI - Surveyors impressed with medical staff involvement. PMID- 15835278 TI - Improve patient flow by reducing bottlenecks. PMID- 15835279 TI - Comparison of clinical and pathological diagnoses in dogs. AB - Clinical and pathological diagnoses were compared in a prospective study of 145 dogs. A diagnostic work up had been performed on all dogs of which 36 (24.8%) died and 109 (75.2%) were euthanatized. In 119 dogs (82.1%) both a clinical and patholical diagnosis was made, in 20 dogs (13.8%) no pathological diagnosis could be made and in 6 dogs (4.1%) no clinical diagnosis was established. In the 119 dogs the agreement level between clinical and pathological diagnosis was scored by the referring veterinarian together with a pathologist. Total agreement was found in 61 cases (51.3%) and disagreement in 31 cases (26.0%). In the remaining cases (27=22.7%) the pathological diagnosis further specified the clinical diagnosis. Consecutive submission appeared difficult to achieve by the participating veterinarians. However, no major differences in agreement level was present between the veterinarian which succeeded in almost consecutive submissions and the other veterinarians. At necropsy 42 cases were diagnosed as neoplasia, of which 52.4% had been diagnosed clinically. As to infectious diseases 55.0% of these diseases diagnosed at necropsy had been diagnosed clinically. In about 20% of the cases the differences were of clinical significance according to the referring veterinarians. In addition, it was indicated by the clinicians that about 50% of the necropsies revealed findings which could amend future patient care. The results of the study stress the relevance of postmortem examination as crucial part of continuing education and of quality monitoring and assurance in veterinary medicine. PMID- 15835280 TI - Neurological syndromes among horses in The Netherlands. A 5 year retrospective survey (1999-2004). AB - The presence of toxins or infectious agents combined with environmental factors in combination with a susceptible host can be the cause for neurological disease in groups of horses. During a 5 year observational period outbreaks of neurological diseases among horses were evaluated. Causes of occurring neurological diseases were equine botulism, lolitrem intoxications, equine herpesvirus type 1-associated myelo(encephalo)pathy, and encephalitis caused by (disseminated) Streptococcus equi subspecies equi infection. This article focuses on the first three syndromes because of their predominant influence on locomotion. The pathogenesis of each disease is presented, followed by a description of a general presentation of the diseases as encountered under Dutch circumstances. PMID- 15835281 TI - The Hazard Analysis Critical Control Point's (HACCP) concept as applied to some chemical, physical and microbiological contaminants of milk on dairy farms. A prototype. AB - Quality management on dairy farms becomes more and more important regarding the different areas of animal health, animal welfare and food safety. Monitoring animals, farm conditions and farm records can be extended with risk identification and risk management. The hazard analysis critical control point's system is useful as an on farm strategy to control the product as well as the production process on the areas of animal health, animal welfare and food safety. This article deals in detail with the question how to develop a qualitative method where risk can be defined as an interaction between probability and impact. Two parts of the production process (milk harvest and treatment of cows) where used as an example how to apply the hazard analysis critical control point's system on chemical, physical and microbiological contaminants of milk. Not just only by summarizing the different critical checkpoints for each area but also by giving them a precise judgement of probability and impact. PMID- 15835282 TI - Huntington's disease like-2: review and update. AB - Huntington's Disease-like 2 (HDL2), like Huntington's disease (HD), is an adult onset, progressive, neurodegenerative autosomal dominant disorder clinically characterized by abnormal movements, dementia, and psychiatric syndromes. Like HD, the neuropathology of HDL2 features prominent cortical and striatal atrophy and intranuclear inclusions. HDL2 is generally rare, accounting for only a few percent of HD-like cases in which the HD mutation has already been excluded. However, the rate is considerably higher among individuals of African ancestry, and is almost as common as HD in Black South Africans. The disorder is caused by a CTG/CAG expansion mutation on chromosome 16q24.3, with normal and expanded repeat ranges similar to HD, and a correlation between repeat length and onset age very similar to HD. Surprisingly, the available evidence suggests that HDL2 is not a polyglutamine disease. Rather, the repeat expansion is located within Junctophilin-3 in the CTG orientation. The phenotypic similarities between HD and HDL2 suggest that understanding the pathobiology of HDL2 may shed new light on the pathogenesis of HD and other disorders of striatal neurodegeneration. PMID- 15835283 TI - Clinical and magnetic resonance imaging manifestations of Holmes tremor. AB - Holmes tremor is a rare symptomatic slow tremor in the proximal parts of the limbs. It may be present at rest or maintenance of a posture, or during the movement of the affected limb. We describe herein three patients of Holmes tremor with possible etiologies of brainstem infarction and head injury. The intervals between the causal events and the appearance of tremor range from 1 month to 12 months. Magnetic resonance imaging studies reveal hypertrophy of the inferior olivary nucleus in all of the three patients, although only one of them has palatal myoclonus. The surface electromyographic recordings reveal characteristic slow oscillation with frequencies of 3.5 to 4.2 Hz. These features suggest that perturbation of the dentato-rubral-olivary circuitry may play a pivotal role for the generation of Holmes tremor. However, no tight correlation is observed between the presence of inferior olivary nuclear hypertrophy and the appearance of symptomatic palatal myoclonus in the current report. PMID- 15835284 TI - Effectiveness of modified constraint-induced movement therapy on upper limb function in stroke subjects. AB - BACKGROUND AND PURPOSE: Of all stroke survivors, more than 50% are left with motor disabilities. Impairment of upper limb movement is a common motor disability. Constraint-Induced Movement Therapy (CIMT) is an intervention which has been used for the treatment of upper extremity motor disabilities in stroke patients. Although CIMT is an effective intervention, a recent survey revealed that this procedure is viewed with apprehension by many clinicians because of concerns about practicality and resource issues. We developed a modified CIMT that reserves the massed training of the affected arm without any physical restriction of the intact one and then used it on our stroke patients. This study was designed to evaluate the effectiveness of this Modified-Constraint-Induced Movement Therapy (m-CIMT). METHODS: Thirty stroke patients were randomly assigned to either an m-CIMT (n=13) or a control group (n=17). Subjects in the m-CIMT group received a 2-week course of m-CIMT. Outcomes were evaluated using the Wolf Motor Function Test (WMFT). RESULTS: After only 2 weeks of training, significant differences (p<0.05) in favor of m-CIMT were found in the following 6 elements of the WMFT: Extend elbow with weight, Lift pencil, Stack checkers, Flip cards, Turn key in lock, and Lift basket. CONCLUSION: The present study shows that our m-CIMT is useful in improving the function of the affected upper extremity in stroke patients. PMID- 15835285 TI - Tuberculous meningitis with initial manifestation of isolated oculomotor nerve palsy. AB - Tuberculous meningitis (TB meningitis) is a subacute meningitis known for its various form of initial manifestations, which often make early diagnosis difficult. The present case report demonstrates a patient with TB meningitis, who had initial manifestation of isolated right oculomotor nerve palsy. High vigilance is needed in diagnosing TB meningitis. A 75 year-old female was hospitalized due to acute onset of right side ptosis. Thorough neurological examination at admission revealed isolated right oculomotor nerve palsy. Brain magnetic resonance imaging and cerebral angiography showed no specific finding. Lumbar puncture was performed two days later due to low grade fever. Cerebrospinal fluid (CSF) study and the polymerase chain reaction on CSF confirmed the diagnosis of TB meningitis. Because TB meningitis is a chronic disease, cranial nerve palsies are common manifestations. This report suggests that TB meningitis should be a disease of differential diagnosis for isolated oculomotor nerve palsy. PMID- 15835286 TI - Fatal meningoencephalitis caused by disseminated strongyloidiasis. AB - A middle-aged aboriginal man with a history of alcoholism and gastrectomy was diagnosed as having bacterial meningoencephalitis based on the typical clinical manifestations, laboratory findings, and treatment responses. During the recovery stage, he developed consciousness disturbance, seizures, severe diarrhea, and respiratory failure that led us to search for other possibility of the diagnosis. The eosinophilia and repeated stool examinations helped us to make the diagnosis of disseminated strongyloidiasis. In this patient the initial bacterial meningitis was followed by S. stercoralis hyperinfection. Despite treatment with strong antimicrobial agents, the patient died. This case could serve as a reminder to physicians to be alert for strongyloidiasis superimposed on bacterial meningitis. PMID- 15835287 TI - Recent advance in immunological tests in paraneoplastic neurological syndrome. AB - Paraneoplastic neurological syndromes are uncommon, however; their diagnosis is of major practical importance. Any portion of the nervous system may be involved in paraneoplastic syndromes. There is increasing evidence that the pathogenesis of many paraneoplastic neurological syndromes appears to be an immune reaction against antigen shared by the cancer and the nervous system. The identification of antibodies in the serum or cerebrospinal fluid in the central nervous system of paraneoplastic syndrome patient confirms the clinical diagnosis of paraneoplastic syndrome, and allows early identification of an underlying tumor at a stage when it is localized and more amenable to treatment. Cancer therapy (surgery, radiotherapy, chemotherapy) seems to be the most efficient treatment for the paraneoplastic neurological symptoms. Immunomodulatory therapy (intravenous immunoglobulin, plasmapheresis, immunosuppression) can halt or even reverse the neurological syndrome. The recent advances in understanding of the autoimmune pathology of these disorders should lead to more effective treatment options. PMID- 15835288 TI - Exertional but not postural headache resulting from spontaneous intracranial hypotension. PMID- 15835289 TI - [Falls in peripheral neuropathy]. AB - Falling accidents are predisposing factors in worsening the quality of life. Causes of falls included gait and balance problems, visual disorder, cognitive impairment and drugs. However, falls in neuropathic patients are not often investigated. We studied the actual condition and causes of falls associated with neuropathy. One hundred and ninety-three patients (122 males and 71 females, ages 57 +/- 15 years) with neuropathy were selected for evaluation. In all patients, more than half a year had passed since the onset of symptoms and the present condition was considered comparatively stable. We divided the patients into 2 groups: falling or non-falling. Patients experiencing accident accompanied by external injury once or more in the past year, or patients who answered "often fell" (1 time or more per month) on flat ground or in their residence comprised the fall group. The severity of disease was evaluated by modified Rankin Scale (0 to 5). The relationship between falls and neuropathic symptoms (proximal muscle weakness, distal muscle weakness, aching or numbness, and the position and vibration sensory loss) was statistically examined. The distribution of patients in the fall group according to modified Rankin Scale of neuropathy was scale 5: 0/0 (0%), scale 4: 5/36 (14%), scale 3: 24/72 (33%), scale 2: 7/56 (13%), and scale 1: 1/19 (5%). There were six fractures among all patients. Regarding the etiology, the fall group showed a high level of axonal neuropathies (44%). On analysis according to condition and symptoms in the patients with a score of 2, 3, or 4 who demonstrated a high rate of falls, there was a strong correlation between deep sensory loss and fall (p < 0.05). However, as independent factors, there were no correlations between falling and proximal muscle weakness, distal muscle weakness or aching numbness, respectively. We thought it necessary to add ataxic gait resulting from deep sensory loss to one of the fall risks. PMID- 15835290 TI - [A case of multiple sclerosis with bilateral useless hand syndrome as a main clinical feature]. AB - A 41-year-old, right-handed man was admitted to our hospital on September 12, 2002, due to progressive clumsiness in both hands. The patient had been diagnosed as having multiple sclerosis three years prior to admission. He noticed difficulty in manipulating objects three months before admission. Cervical T2 weighted MRI showed a high signal intensity at the level of C3-4 which was enhanced on T1-weighted image with gadolinium. On admission, neurological examinations revealed impairment of dexterity, deep sensory disturbance, and astereognosis in both hands. The clumsiness of complex finger movements was predominant on the left side, and was exaggerated with the eyes closed in association with pseudoathetosis. After steroid therapy, his clumsy hands improved gradually. This type of clumsiness in multiple sclerosis had been described as useless hand syndrome by Oppenheim. In accordance with our case, useless hand syndrome has been reported to arise from high cervical (C2-4) lesions, mainly involving the posterior cord ipsilateral to the clumsy hand. Although the majority of reported cases with useless hand syndrome had other neurological complications, such as hemiparesis, tetraparesis, and truncal ataxia, our patient exhibited a pure form of useless hand syndrome. In addition, useless hand syndrome is usually unilateral, and bilateral useless hand syndrome is very rare. Clumsiness of fine finger movements with astereognosis in our patient is similar to numb clumsy hands or limb-kinetic apraxia due to cervical spondylosis or postcentral gyrus lesion, respectively. This indicates an important role of the high cervical posterior cord in conveying a kinesthetic sense necessary to guide fine finger movements. It should be kept in mind that high cervical lesions in multiple sclerosis causes clumsy hands mimicking limb kinetic apraxia. PMID- 15835291 TI - [A case of migraine with aura who showed no visual symptom]. AB - A 22-year-old man experienced a severe migraine on left side of the head with an aura including aphasia and a numbness on the right hand. A migraine with an aura of sensory disturbance on the both hands recurred immediately after cerebral angiography. Further examinations including MRI and angiography did not reveal any evidence of cerebrovascular diseases. Aphasic migraine was diagnosed in this patient. At all attacks, this patient never experienced visual aura which was common in the previous reports of migraine. PMID- 15835292 TI - [A pedigree of Charcot-Marie-Tooth disease type 4F (Periaxin mutation)]. AB - We report a 51-year-old man genetically diagnosed as Charcot-Marie-Tooth disease type 4F. The patient was the first child of healthy, consanguineous parents. He had two sisters and one of them showed similar but milder symptoms. He had gait disturbance since childhood. Then he noticed muscle weakness of his hands at the age of early forties, and more difficulties in gait at the age of late forties. On examination at age 51, he showed absence of all deep tendon reflexes, weakness of the hand and distal leg muscles, pes cavus and decreased sensitivity to touch and vibration in the lower extremities. Electrophysiological studies of the median nerve showed delayed motor nerve conduction velocity and undetectable sensory nerve action potentials. The histology of his sural nerve revealed moderate loss of large myelinated fibers and the diameters of residual fibers shifted to small shown as size-frequency histogram. Many fibers are thinly myelinated and some of the Schwann cells looked as wrapping around the myelinate fibers with their processes. On gene analyses, we identified an Arg 1070 Stop homozygous mutation in the Periaxin, known to be a causative gene for CMT type 4F. Based on these observations, we emphasized that broad genetic analyses are necessary for diagnosis of CMT disease, including so far unidentified mutations among the Japanese populations. PMID- 15835293 TI - [Dissociated micturitional disturbance in a patient with cervical spondylotic myelopathy]. AB - A 58-year-old man developed dysuria and residual sensation one year after the traffic accident. Half a year later, he developed chest pain and underwent coronary arteriography. Numbness in the left arm, burning sensation below the knees and urinary retention emerged immediately after prolonged neck extention during the examination. He required intermittent catheterization of the bladder because of persistent dysuria. Urodynamic study revealed detrusor-sphincter dyssynergia. The micturitional disturbance improved with the application of a soft cervical collar and a bed rest. MRI of the cervical spine showed spinal cord compression at the C5/6 and a small hyperintensity area on T2WI in the spinal cord at the C5 level. The case suggests that micturitional dysfunction can be severe without remarkable sensorimotor disturbances of the lower extremities in cervical spondylotic myelopathy. PMID- 15835294 TI - [A 95-year-old female with autopsy-proven cerebral necrosis due to candidiasis who developed stroke-like manifestations]. AB - A 95-year-old woman complained of sudden onset of disturbance of consciousness and right hemiparesis on April 20, 2003 and was admitted on the next day. She was drowsy and showed moderate right motor and sensory hemiparesis. The blood laboratory tests showed slight inflammatory reaction. A low density area was found in the left basal ganglia by brain CT, which was also coincided with the high signal region in T2, FLAIR and diffusion-MR images. The MRA of the intracerebral arteries presented no remarkable abnormality. The hemiparesis and impaired consciousness improved partially in the following week. However, she did not fully recover, since aspiration pneumonia and mild generalized inflammation continued. Percutaneous gastrostomy and intravenous hyperalimentation were started to improve her nutrition. The moderate inflammatory state persisted for several weeks. Her blood pressure suddenly fell and she died on June 12. Autopsy showed a mildly brownish and necrotic lesion from the left caudate to the putamen through the internal capsule. There was no liquefaction. On the microscopic examination, the necrosis surrounded by small vessels was consisted of numerous neutrophils and macrophages with pseudohypha and blastospore of candida. Small fragments of fungus were phagocytosed by macrophages. Small abscesses and necrotic foci due to candidiasis were observed in the bladder, kidneys, lungs, myocardium and thyroid gland. In this case, cerebral candidiasis probably occurred via hematogenous dissemination from a primary focus in the urinary tract. The intracerebral arteries revealed rather mild atherosclerotic changes and there was no occlusion by thromboembolism. Intracerebral lesion was diagnosed as candidiasis and there was no cerebral infarction by thromboembolism. If the infection occurred after cerebral infarction, there should not be any inflammatory reaction in the center of necrotic area. There have been few reports of cerebral candidal infection in patients without diabetes mellitus or immunosuppressive conditions. None of them had been diagnosed before death. Caution should be exercised for the presence of systemic candidiasis in elderly patients who are bedridden and with continuous low grade inflammatory reactions. PMID- 15835295 TI - [Portal vein thrombosis associated with hepatic encephalopathy]. AB - A 54-year-old man who had been administered chlormadinone acetate 3 months after prostatectomy for prostate cancer, acutely developed disorientation and memory disturbance. Six days later, he experienced high grade fever and epigastralgia. He was suspected to have hepatic encephalopathy, because the Fischer ratio was low although serum ammonia level remained normal. Further examinations including abdominal echography and CT scan disclosed a thrombus extending from the portal vein to the superior mesenteric vein together with abnormal collateral vessels originating from the portal vein. He was successfully treated with warfarin potassium, urokinase and heparin sodium. It was suggested that the patient developed hepatic encephalopathy due to portal-systemic circulation shunting secondary to portal vein thrombosis. PMID- 15835296 TI - [Acute encephalopathy among patients with renal dysfunction after ingestion of "sugihiratake", angel's wing mushroom--study on the incipient cases in the northern area of Niigata Prefecture]. AB - An outbreak of acute encephalopathy among 6 patients with renal dysfunction after eating "Sugihiratake" mushroom in the northern area of Niigata Prefecture was reported. All of the patients had varying degrees of renal dysfunction, and 3 of them were on dialysis treatment. Patients initially presented with shaking limbs and difficulty in walking. Several days later, disturbances in consciousness developed, which were followed by status epileptics. Four patients were put on mechanical ventilation for seizure control. Two of the 6 cases died. CSF examination showed elevated protein without pleocytosis; bacterial, rickettsial and viral screenings was negative. Brain CT and MRI studies showed edema in the claustrum and external capsule, and in the white matter of the base of the frontal, parietal, and temporal lobes. No common drug administration was recognized. All of the patients had a history to have ingested "Sugihitarake" mushroom (angels' wing mushroom, pleurocybella porrigens) in varying quantities and frequencies prior to the onset of the illness. This mushroom may have induced toxic encephalopathy although no similar case have been reported in the past. PMID- 15835297 TI - [A case of unilateral ischemic stroke with ipsilateral limb-kinetic apraxia]. AB - A 74-year-old man developed aphasia, weakness of the right upper extremity and left limb-kinetic apraxia. Brain MRI showed an infarct in the area supplied by the left middle cerebral artery. Cerebral angiography revealed high grade stenosis with plaques of bilateral proximal internal carotid arteries. The carotid endarterectomy of the left carotid artery was performed three months later. After this operation his left limb-kinetic apraxia improved. We considered transhemispheric diaschisis, callosal apraxia or diagonistic dyspraxia as a possible cause of this rare symptom. PMID- 15835298 TI - [Coexistance of cluster headache and hemicrania continua: a case report]. AB - We reported a 36-year-old man, who suffered from cluster headache (CH) associated with hemicrania continua (HC). The continuous, dull or pressure-type headache appeared on the same side of the CH during the third month of a prolonged cluster period, and fluctuated in the severity of pain. This headache was aggravated when the CH was ameliorated by the administration of lithium carbonate. This converse relationship between CH and HC persisted during an on-off trial of the lithium carbonate, and the HC was exacerbated again after the complete cessation of CH. Retrobulbar pain and nasal congestion were present as components of HC similarly to CH, but they subsided gradually and the pressure-type vascular headache over the temporal area predominated later. The continuous headache lasted more than 3 months, and responded significantly to the indomethacin at a dose of 75mg/d. The clinical course of this patient suggests that HC and CH have a common pathomechanism including hyperactivation of the trigemino-vascular reflex, and may be different in the involvement of other central pathway of pain generation. Indomethacin may deserve consideration for the treatment of continuous headache that appears during an atypical course of other primary headaches. PMID- 15835299 TI - [A case of encephalitis-type encephalopathy related to Pleurocybella porrigens (Sugihiratake)]. AB - A 65-year-old man who had been on hemodialysis for chronic renal failure was well until 7 days after ingestion of Pleurocybella porrigens (Sugihiratake) when he developed a loss of consciousness and status epilepticus of generalized tonic and clonic seizures. High fever and high CRP were followed and cerebrospinal fluids showed marked pleocytosis with polynuclear cell predominance. Brain MRI showed diffuse lesions in the basal ganglia and multiple ringed lesions in cerebral cortex. Glucocorticoid therapy was effective for clinical improvement. This case showed the presence of encephalitis-type of encephalopathy related to Pleurocybella porrigens (Sugihiratake). PMID- 15835300 TI - Herpes simplex virus encephalitis presenting with cerebral infarction-like signs and neuroimages. AB - A patient with an atypical presentation of herpes simplex virus (HSV) encephalitis mimicking acute cerebral infarction was reported. A 48-year-old man developed left-sided hemiparesis, convulsive seizures, and loss of consciousness. Brain magnetic resonance imaging revealed high intensity areas in the right frontal to parietal lobes on T2-weighted and diffusion-weighted images. Soon after admission with suspected cerebral infarction of the right middle cerebral artery region, the patient had high fever with frequent seizures and severe loss of consciousness. Laboratory findings including cerebrospinal fluid established a diagnosis of HSV encephalitis, and a state of apalic syndrome persisted despite aggressive antiviral therapy. PMID- 15835301 TI - [Sex identification from DNA in old teeth]. AB - The sex determination was carried out on 80 fresh and 15 old teeth by amplifying sex chromosome specific sequences with the polymerase chain reaction. The DNA content in the tooth significantly decreases with aging. There was no correlation between days after evulsion and the amounts of DNA extracted from fresh teeth that had been preserved less than 186 days. The sex determination based on fresh teeth was successful using multi locus markers, DYZ-1 and DYZ-3 in combination with DXZ-1. However, amelogenin and pseudoautosomal boundary, both that are single locus markers and specific for both sex chromosomes with different lengths, could not be detected in three samples, of which DNA contents were extremely low. However, the sex determination by amelogenin amplified with fluorescent probes was possible in these three samples. We also determined sexes of 30 old specimens (15 teeth and 15 bones) from 15 human skulls using sex chromosomes locus markers. Prior to molecular sex determination, two forensic specialists determined the sex of the skull morphologically. From the 15 skulls, sex identification using multi locus marker (DYZ-1 or DYZ-3) was possible for 12 of 15 teeth and 7 of 15 bones. The sex was successfully determined from 11 teeth and 9 bones by amplification of the amelogenin locus. However, the coincidence rate of the molecular test with morphological examination was < 20%. In conclusion, sex determination on the fresh tooth would be successful using any sex chromosome marker. However, in cases on samples that have spent considerable years in the ground, pollution and putrefaction, especially, by humicolous, must be considered. Thus, sex determination by DNA testing should be regarded as accurate, when the results from two or more molecular markers are coincident. Hokkaido J Med Sci 80(2), 191-199, 2005 PMID- 15835303 TI - Bone marrow transplantation and medulla architecture reconstitution in the murine thymus. PMID- 15835302 TI - [In silico and systematic molecular evolutionary analysis of human SIAH1b inactivation]. AB - "Seven in absentia homolog" (SIAH) is E3 ubiquitin ligase gene. It is reported that mice have three SIAH homolog genes, Siah1a, Siah1b and Siah2, while men have SIAH1 and SIAH2 genes. It has recently been reported that, among these genes, mouse Siah1b gene is a direct target of p53. However, the nature of human "SIAH1b" gene remains unknown. In this study, we identified it on chromosome X by using a computed searching methods (in silico cloning). Using cDNA from human tissues, we attempted to amplify this region by PCR, but failed to amplify it. By analyzing the structure of human "SIAH1b" gene, we found that Alu was inserted immediately before its coding region, and that this gene had a number of base substitutions in its coding region, compared to its homolog genes in mouse or man. From these findings, we suggest that human "SIAH1b" gene had been inactivated by insertion of Alu, and estimate that the inactivation occurred about 30-58 million years ago from the substitution rates of base during evolution. PMID- 15835304 TI - [Introduction to chromatin choreography]. PMID- 15835305 TI - [Chromatin dynamics in DNA replication and gene transcription]. PMID- 15835306 TI - [Role of DNA polymerase a in genome stability]. PMID- 15835307 TI - [Physiological functions of the acidic molecular chaperone Nap1]. PMID- 15835308 TI - [Nucleosome assembly: chromatin and epigenetics]. PMID- 15835309 TI - [Structure and function of centromere chromatin: CENP-A defines the centromere region]. PMID- 15835310 TI - [Role of ATM-related proteins in the assembly of telomere replication proteins]. PMID- 15835311 TI - [View of Topo II as a highly dynamic protein]. PMID- 15835312 TI - [Aurora dance on the mitotic ballet]. PMID- 15835313 TI - [Centrosome as an organizing center of intracellular information: insights viewed from the spindle pole of budding yeast]. PMID- 15835314 TI - Preconceptions in the nurse-client relationship. AB - Nursing theorist, Hildegard Peplau (1952) has identified the concept of preconceptions as critical in the development of the therapeutic nurse-client relationship. Although stereotypes exist for both nurses and chronic psychiatric clients, very little research has been reported on the preconceptions nurses and psychiatric clients have of each other. This investigation utilized non probability, purposive sampling of 20 newly formed nurse-client dyads within programmes serving a chronically mentally ill population in Canada. Subjects were asked to give descriptions of each other. Semantic differentials based on this feedback were then developed and administered to 124 nurse-client dyads. Clients' statements generally evaluated their nurses positively. The generally positive views expressed by nurses and clients did not reflect public stereotypes for either group. The preconceptions the clients had of their nurses, and nurses had of their clients were related to both the quality of the emerging relationship (task, bond and goals) and the duration of the orientation phase. The preconceptions were virtually unchanged over the initial 6 months of the relationship. PMID- 15835315 TI - Concept and models for the study of caregiver burden. AB - Families caring for a mentally ill member experience various degrees of burden. The concept of burden has been studied over the centuries and is seen primarily as an obligation which entails a cost. In very few cases burden has been seen positively. The present article explores the concept of burden as it is presented in the literature, addresses the categories of strain that families caring for mentally ill relatives experience, and presents four models for the study of caregiver burden. PMID- 15835316 TI - Siblings and eating disorders: a phenomenological perspective. AB - The purpose of this study was to develop an increased awareness of the sibling experience in families where one child has an eating disorder. Using a phenomenological approach, five female adolescents whose sisters had been diagnosed with anorexia nervosa each participated in two open-ended, unstructured interviews, which were coded and analysed using methods consistent with phenomenological research. Two overarching constructs evolved, which subsumed five major themes. All siblings (1) described intense and conflicted emotions, and (2) experienced their sisters' illness as a pervasive phenomenon in their lives. In this context, perspective of the illness, disruption of intra- and extrafamilial relationships, role strain, special status awarded the anorectic sister, and coping with the illness were significant themes which emerged for participants. Findings support the need for nurses to be involved in family focused health care practice, and to be cognizant of the impact on siblings in families where one member's illness is a factor. PMID- 15835317 TI - Factors influencing commitment to abstinence from alcohol: a model grounded from client perceptions of disulfiram therapy. AB - This paper describes the findings of a qualitative research study aimed at eliciting the perspectives of problem drinkers concerning the drug disulfiram (Antabuse). A grounded theory model of Commitment to Abstinence, which was generated from the study data, is presented. The influence of disulfiram upon this model is examined via the perspectives of disulfiram users. It is suggested that the drug could be used in a flexible way with some clients to their advantage. The significance of the study and the derived model for mental health nursing is discussed, the value of the model as an assessment tool is explored and other more generic applications are suggested. PMID- 15835318 TI - The experience of special hospital patients in regional secure units. AB - Regional secure units provide one of the main routes of release for special hospital patients; a route that in many cases is unsuccessful as the patient is subsequently readmitted to the special hospital. Processes underlying this were investigated in this UK study using a grounded theory approach. The interview data obtained in 1993 from a sample of 14 readmitted special hospital patients were analysed and five explanatory categories were generated. This paper concentrates on descriptive data from two categories. One describes negative experiences of a group of patients in regional secure units; an experience often considered a 'backward step'. The second data category identifies four principal reasons for readmission; one reason, inappropriate sexual behaviour, was of particular concern due to its frequency and the possibility that it resulted from the special hospital practice of gender segregation. PMID- 15835319 TI - A review of the Report on mental health nursing in England: working in partnership. AB - We invited Professor Smith to provide a personal perspective on the context and relevance of the first major review of psychiatric and mental health nursing in England since 1968. The British government published a report of the review in 1994 and the chairman of the steering group summarized the main issues and characterized its findings in this Journal (1, 41-44). In this paper, Professor Smith establishes the provenance of the Review of Mental Health Nursing. Consideration is given to the Report's recommendations in an effort to understand reaction to it. Finally, ideas are suggested to take forward the Report's recommendations, using the Report as a useful tool to progress psychiatric and mental health nursing. We need to be aware that without comment from the profession this Report will cease to be of interest or of relevance. PMID- 15835320 TI - The efficacy of cognitive-behavioural pychotherapy for hypochondriasis: a pilot investigation. PMID- 15835321 TI - Comparing one aspect of the care of elderly patients with dementia in psychogeriatric and continuing care settings. PMID- 15835322 TI - Getting a fair hearing: a small step towards client empowerment. PMID- 15835323 TI - Living with ambiguity. PMID- 15835324 TI - Looking back at the past four years of Labour rule. PMID- 15835325 TI - Voice of older people's nursing. PMID- 15835326 TI - Needlestick injuries. PMID- 15835327 TI - Healing hearts. PMID- 15835328 TI - The role of angioplasty in acute myocardial infarction. AB - Most patients in the UK with an acute myocardial infarction (MI) are treated with thrombolysis. This article discusses the role of primary coronary angioplasty as the first-line treatment for acute MI. PMID- 15835329 TI - Schistosomiasis. PMID- 15835330 TI - Ipratropium bromide. PMID- 15835331 TI - Choice in the diet of people with learning difficulties. AB - Many adults with learning disabilities experience difficulties communicating food preferences because they may have little opportunity to do so. This article discusses how health care professionals can enable people with learning difficulties to exercise choice about food and how those choices can be recognised. PMID- 15835332 TI - Advances in prevention and treatment of stroke and TIA. AB - There is increasing recognition of the importance of stroke prevention and effective treatment and management of people who have had a stroke or transient ischaemic attack. This has led to the development of a range of service developments across all health care settings, which are bringing exciting and challenging opportunities for nurses. This article highlights key clinical developments in stroke prevention and treatment, and outlines some of the roles open to nurses who wish to become involved in stroke care provision. PMID- 15835333 TI - Improving care for patients with nausea and vomiting. PMID- 15835334 TI - The role of relationships in effective clinical supervision. AB - AIM: To investigate the nature of clinical supervision. METHOD: Within the four cycle action research approach qualitative data was collected using a range of methods and analyses. RESULTS: Several insights were gained into the nature of clinical supervision and the nature of change required to support its introduction. One such insight, was the importance of relationship development. CONCLUSION: Relationship development is an essential component of effective clinical supervision. PMID- 15835335 TI - Lung cancer: an update on current diagnostic techniques and treatment. AB - National Clinical published Lung cancer is the second most commonly diagnosed cancer in the UK. In 2000 there were 38,410 new cases, accounting for 14 per cent of all cancers diagnosed (Cancer Research UK, 2004). While the incidence of lung cancer is less than that for breast cancer, lung cancer deaths account for more than one-fifth of all cancer deaths. In 2002 this amounted to 33,600 lung cancer deaths, or 22 per cent of all cancer deaths (Cancer Research UK, 2004). PMID- 15835336 TI - NICE guidelines for the diagnosis and treatment of lung cancer. AB - Lung cancer is the most common cause of cancer death, accounting for nearly 30,000 deaths in England and Wales in 2002 (DoH, 2005). On February 23 this year new guidelines for the diagnosis and treatment of lung cancer were published by the National Institute for Clinical Excellence (NICE, 2005). The guidelines offer best practice advice on the care of adults who are suspected of having, or are diagnosed with, lung cancer (Box 1). There are implications for lung cancer clinical nurse specialists in several sections of the guidance, some highlighted as a 'key priority' for implementation. PMID- 15835337 TI - Assessment and nursing care of the patient with dyspnoea. AB - Breathlessness is a subjective experience, which has been described as an unpleasant or uncomfortable awareness of breathing, or of the need to breathe (Gift, 1990). Patients experiencing breathlessness are often described as being dyspnoeic, or having dyspnoea. This is derived from the Greek word, which, when translated, means 'difficulty in breathing'. PMID- 15835338 TI - Home non-invasive ventilation: a brief guide for primary care staff. AB - An increasing number of patients with a wide range of respiratory problems are using non-invasive ventilation (NIV) at home to optimise their respiratory function. It has been estimated that in 1999 between 2,500 and 3,000 people in the UK (5 per 100,000 head of population) were using domiciliary NIV (Leger, 2001). As a consequence, health professionals working in the community are now more likely to encounter patients using this type of ventilation. PMID- 15835339 TI - How to engage in research. PMID- 15835340 TI - Helping people back into society. PMID- 15835341 TI - Not even Dr. Evil likes MRSA. PMID- 15835342 TI - [Diabetic metabolic emergencies]. AB - Diabetic ketoacidosis and hyperglycaemic hyperosmolar syndrome are two acute complications of diabetes that may result in high mortality. Effective standardized treatment protocols, as well as prompt identification and treatment of the precipitating cause, are important factors affecting outcome. The authors review the pathomechanism, clinical features, complications and current recommendations for management of ketoacidosis and hyperglycaemic hyperosmolar syndrome. In patients with type 2 diabetes may develop anaerobic (type A) lactic acidosis from tissue hypoxia, while aerobic (type B) lactic acidosis is a rare complication of biguanide therapy if contraindications to metformin are observed. Mortality remains very high, especially when serious comorbidities are present. Hypoglycaemia is a major factor preventing patients with both type 1 and type 2 diabetes from achieving near-normal plasma glucose targets. The authors summarize clinical presentation and therapeutic possibilities of hypoglycaemia on the basis of the literature. PMID- 15835343 TI - [Proarrhythmic (torsadogenic) effects of QT-prolonging non-antiarrhythmic drugs]. AB - The authors review the rapidly expanding knowledge relating to the arrhythmogenic (torsadogenic) effects of non-antiarrhythmic drugs that lengthen ventricular repolarization (i.e. the duration of electrocardiographic QT interval). After outlining the ECG characteristics and electrophysiological mechanism of drug induced long-QT syndrome and torsades de pointes ventricular tachycardia, they discuss the concept of repolarization reserve and its role in arrhythmogenesis. Information is provided on the cardiac ion channel background of the proarrhythmic action of QT-prolonging non-antiarrhythmic drugs. The authors present pharmacoepidemiological data on the clinical importance and incidence of this peculiar adverse drug reaction that can in certain cases cause sudden arrhythmic death. They draw attention to the predisposing factors of torsades de pointes ventricular tachycardia, and list those groups of drugs and individual agents that have QT-lengthening and torsadogenic side-effects. PMID- 15835344 TI - [Second cancers in hematologic malignancies (epidemiologic observations from a 20 year period)]. AB - INTRODUCTION: The incidence of malignant tumours have increased steadily worldwide. Prior reports indicate that patients with some hematologic malignancies (for example chronic lymphocytic leukemia, non-Hodgkin's lymphoma) may be at increased risk of second neoplasms. AIM: The aim of the authors was to explore the possible association between hematologic malignancies and subsequent solid tumours. PATIENTS, METHODS: Between January 1, 1983 and December 31, 2002, in the county of Szabolcs-Szatmar-Bereg 151 cases with both malignant hematologic diseases and cancers were registered by the authors. In their 60 patients (50 with lymphoid and 10 with myeloid malignancies) the first tumour was the hematologic malignancy. Among these cases the number of second cancers was 64. RESULTS: The most common kind of second tumours was lung cancer. There was a significant connection between lymphoid malignancies and second cancers as compared to myeloid malignancies and subsequent cancers (p < 0.05). CONCLUSIONS: Analysing the epidemiologic data the authors established the following conclusions: the association of second cancers with hematologic malignancies in most of the cases is not accidental. The age of patients with second cancers seemed to be not too important, but it was of crucial importance in patients with Hodgkin's disease and non-Hodgkin's lymphoma. The role of immunodeficiency in the development of second cancers may be important in patients with Hodgkin's disease, non-Hodgkin's lymphoma and chronic lymphocytic leukemia (the number of second cancers in their patients with multiple myeloma and hairy cell leukemia was too small do draw a conclusion). PMID- 15835345 TI - [Femoral press-fit fixation in anterior cruciate ligament reconstruction: postoperative histological examination of bone block incorporation in pigs]. AB - INTRODUCTION: The histological properties of the femoral press-fit fixation in anterior cruciate ligament reconstruction have not been extensively examined. The purpose of the study was to analyse the histological properties of the bone patellar tendon graft in the bone tunnel area during the early stages of healing in an animal model. Furthermore the authors wanted to draw conclusions for the postoperative rehabilitation program on the basis of their data. MATERIAL AND METHODS: ACL reconstruction was performed in pigs using femoral press-fit fixation. After 3 and 6 weeks the incorporation of the bone block in the femur in 2 pigs was histological and radiological evaluated. RESULTS: After 3 and 6 weeks the examination showed almost similar histology on the bony side the grafts except (1) a higher amount of fibrotic tissue in-between the lamellae of the inserted patellar bone after 6 weeks, (2) the presence of direct osseous unions between the two lamellar systems and (3) thicker lamellae at the base than toward the condylar tissue. CONCLUSIONS: Concerning the progression of graft healing by histological findings our study showed, that after 6 weeks the femoral press-fit fixation appear to have good incorporation after ACL reconstruction. PMID- 15835346 TI - [Struggle against tuberculosis in Hungary during the First World War, especially in the military medical institute of Besztercebanya and Rozsahegy]. PMID- 15835347 TI - Optical micromanipulations inside yeast cells. AB - We present a combination of nonlinear microscopy and optical trapping applied to three-dimensional imaging and manipulation of intracellular structures in living cells. We use Titanium-sapphire laser pulses for nonlinear microscopy of the nuclear envelope and the microtubules marked with green fluorescent protein in fission yeast. The same laser source is also used to trap small lipid granules naturally present in the cell. The trapped granule is used as a handle to exert a pushing force on the cell nucleus. The granule is moved in a raster-scanning fashion to cover the area of the nucleus and hence displace the nucleus away from its normal position in the center of the cell. Such indirect manipulations of an organelle (e.g., nucleus) can be useful when direct trapping of the chosen organelle is disadvantageous or inefficient. We show that nonlinear microscopy and optical manipulation can be performed without substantial damage or heating of the cell. We present this method as an important tool in cell biology for manipulation of specific structures, as an alternative to genetic and biochemical methods. This technique can be applied to several fundamental problems in cell biology, including the mechanism of nuclear positioning and the spatial coordination of nuclear and cell division. PMID- 15835348 TI - Optimized birefringence changes during isolated nerve activation. AB - Single trial, birefringence signals associated with action potentials from isolated lobster nerves were optimized with high-intensity light-emitting diodes (LEDs) and glass polarizers. The narrow spectral output of the LEDs allowed us to select specific wavelengths, increasing the effectiveness of the polarizers and minimizing the stray light in the system. The LEDs produced intensity profiles equivalent to narrowband filtered 100-W halogen light, and birefringence signals were comparable or superior in size and clarity to halogen lamp recordings. The results support a direct correlation between signal size and polarizer extinction coefficient. Increasing the sensitivity of birefringence detection through the use of LED light sources could ameliorate noninvasive brain imaging techniques that employ fast optical consequences associated with action potential propagation. PMID- 15835349 TI - Low-cost, scalable laser scanning module for real-time reflectance and fluorescence confocal microscopy. AB - We present a low-cost, high-speed, retrofitted laser scanning module for microscopy. The cage-mounted system, with various available fiber-coupled sources, offers a real-time imaging alternative to costly commercial systems with capabilities for conventional or confocal reflectance and fluorescence applications as well as advanced laser scanning microscopy implementations. Reflectance images of a resolution target and confocal images of fluorescent polystyrene beads are presented for system characterization. Confocal fluorescence image stacks of T84 epithelial cancer cells are presented to demonstrate application to biological studies. This laser scanning module is a flexible, scalable, high-speed alternative to commercial laser scanning systems suitable for applications requiring a simple imaging tool and for teaching laboratories. PMID- 15835350 TI - Rapid optical coherence tomography and recording functional scattering changes from activated frog retina. AB - Optical coherence tomography (OCT) has important potential advantages for fast functional neuroimaging. However, dynamic neuroimaging poses demanding requirements for fast and stable acquisition of optical scans. Optical phase modulators based on the electro-optic effect allow rapid phase modulation; however, applications to low-coherence tomography are limited by the optical dispersion of a broadband light source by the electro-optic crystal. We show that the optical dispersion can be theoretically estimated and experimentally compensated. With an electro-optic phase modulator-based, no-moving-parts OCT system, near-infrared scattering changes associated with neural activation were recorded from isolated frog retinas activated by visible light. PMID- 15835351 TI - Spectroscopic imaging for detection of ischemic injury in rat kidneys by use of changes in intrinsic optical properties. AB - It is currently impossible to consistently predict kidney graft viability and function before and after transplantation. We explored optical spectroscopy to assess the degree of ischemic damage in kidney tissue. Tunable UV laser excitation was used to record autofluorescence images, at different spectral ranges, of injured and contralateral control rat kidneys to reveal the excitation conditions that offered optimal contrast. Autofluorescence and near-infrared cross-polarized light-scattering imaging were both used to monitor changes in intensity and spectral characteristics, as a function of exposure time to ischemic injury. These two modalities provided different temporal behaviors, arguably arising from two different mechanisms providing direct correlation of intrinsic optical signatures to ischemic injury time. PMID- 15835352 TI - Spectrally programmable light engine for in vitro or in vivo molecular imaging and spectroscopy. AB - A spectrally and temporally programmable light engine can create any spectral profile for hyperspectral, fluorescence, or principal-component imaging or with medical photonics devices employing spectroscopy, microscopy, and endoscopy. Multispectral imaging feasibility was demonstrated by capturing nine images at wavelengths from 450 to 650 mm (25-nm FWHM) with a CCD-camera-equipped bronchoscope coupled to the light engine. Selected wavelength regions were combined to produce a color endoscopy image. PMID- 15835353 TI - Characterization of dentin, enamel, and carious lesions by a polarization sensitive optical coherence tomography system. AB - Enamel and dentin are the primary components of human teeth. Both of them have a strong polarization effect. We designed a polarization-sensitive optical coherence tomography (PSOCT) system to study the spatially resolved scattering and polarization phenomena of teeth. The system is constructed in free space to avoid the complexity of polarization control in fiber-based PSOCT. The structural features of enamel were evaluated in five human teeth that had no visible evidence of caries. The teeth were subsequently sectioned in mesial distal orientation and coronal orientation. Then the structural aspects of dentin were evaluated. OCT images were made of the mantel dentin near the dentin-enamel junction. Five teeth with interproximal and occlusal caries were also studied. With two channel and phase-retardation images, PSOCT provided better functional contrast and more detailed structural information than conventional OCT. For a better description of the measured PSOCT data, we classify these features by two types, i.e., the local textural features and the global structural features. This study indicates that PSOCT has the potential to be a powerful tool for research of dental formation and caries diagnosis. PMID- 15835354 TI - Modeling anisotropic light propagation in a realistic model of the human head. AB - A Monte Carlo model capable of describing photon migration in arbitrary three dimensional geometry with spatially varying optical properties and tissue anisotropy is presented. We use the model to explore the effects of anisotropy for optical measurements of the human head. An anisotropic diffusion equation that corresponds to our Monte Carlo model is derived, and a comparison between the Monte Carlo model and the diffusion equation solution with finite elements is given. PMID- 15835355 TI - P(N) approximation for frequency-domain measurements in scattering media. AB - Presented here are expressions for the P(N) approximation for light propagation in scattering media in the frequency domain. To elucidate parametric dependencies, the derivation uses normalization of the resulting expressions to either the total interaction coefficient or the reduced total interaction coefficient. For the latter case, a set of reduced phase function coefficients are introduced. Expression of the P(N) approximation as a conventional eigenvalue problem facilitates computation of the eigenvalues or attenuation coefficients. This approach is used to determine the attenuation coefficients in the asymptotic regime over the full values of the scattering albedo and reduced scattering albedo (0 to 1) and all positive values of the asymmetry factor (0 to 1). Frequency-domain measurements yield a sensitivity to turbid media optical properties for reduced scattering albedos as small as 0.2. P(N) calculations are used to assess the magnitude of errors associated with the P1 and P3 approximations over a range of scattering albedo, phase function, and modulation frequency. PMID- 15835356 TI - Sources of scattering in cervical tissue: determination of the scattering coefficient by confocal microscopy. AB - Most models of light propagation through tissue assume that the scattering properties of various tissue layers are the same. We present evidence that the scattering coefficient of cervical epithelium varies by a factor of 3 within the epithelium owing to variations in nuclear density and to the presence of keratin. We estimated the scattering coefficient from regions of normal and precancerous cervical epithelium by fitting reflectance measurements from confocal images to an exponential function of depth based on Beer's law of attenuation. The results suggest that the normal cervix is characterized by highly variable scattering in the superficial epithelium, low scattering in the intermediate epithelium, and high scattering in the basal and stromal regions. In high-grade dysplasia, high scattering from high-density nuclei is observed throughout the entire epithelium. PMID- 15835357 TI - Diffuse optical tomography with spectral constraints and wavelength optimization. AB - We present an algorithm that explicitly utilizes the wavelength dependence of tissue optical properties for diffuse optical tomography. We have previously shown that the method gives superior separation of absorption and scattering. Here the technique is described and tested in detail, and optimum wavelength sets for a broad range of chromophore combinations are discovered and analyzed. PMID- 15835358 TI - Practical and adequate approach to modeling light propagation in an adult head with low-scattering regions by use of diffusion theory. AB - A practical and adequate approach to modeling light propagation in an adult head with a low-scattering cerebrospinal fluid (CSF) region by use of diffusion theory was investigated. The diffusion approximation does not hold in a nonscattering or low-scattering regions. The hybrid radiosity-diffusion method was adopted to model the light propagation in the head with a nonscattering region. In the hybrid method the geometry of the nonscattering region is acquired as a priori information. In reality, low-level scattering occurs in the CSF region and may reduce the error caused by the diffusion approximation. The partial optical path length and the spatial sensitivity profile calculated by the finite-element method agree well with those calculated by the Monte Carlo method in the case in which the transport scattering coefficient of the CSF layer is greater than 0.3 mm(-1). Because it is feasible to assume that the transport scattering coefficient of a CSF layer is 0.3 mm(-1), it is practical to adopt diffusion theory to the modeling of light propagation in an adult head as an alternative to the hybrid method. PMID- 15835359 TI - Improved accuracy of reconstructed diffuse optical tomographic images by means of spatial deconvolution: two-dimensional quantitative characterization. AB - Systematic characterization studies are presented, relating to a previously reported spatial deconvolution operation that seeks to compensate for the information-blurring property of first-order perturbation algorithms for diffuse optical tomography (DOT) image reconstruction. In simulation results that are presented, this deconvolution operation has been applied to two-dimensional DOT images reconstructed by solving a first-order perturbation equation. Under study was the effect on algorithm performance of control parameters in the measurement (number and spatial distribution of sources and detectors, presence of noise, and presence of systematic error), target (medium shape; and number, location, size, and contrast of inclusions), and computational (number of finite-element-method mesh nodes, length of filter-generating linear system, among others) parameter spaces associated with computation and the use of the deconvolution operators. Substantial improvements in reconstructed image quality, in terms of recovered inclusion location, size, and contrast, are found in all cases. A finding of practical importance is that the method is robust to appreciable differences between the optical coefficients of the media used for filter generation and those of the target media to which the filters are subsequently applied. PMID- 15835360 TI - Design and implementation of dynamic near-infrared optical tomographic imaging instrumentation for simultaneous dual-breast measurements. AB - Dynamic near-infrared optical tomographic measurement instrumentation capable of simultaneous bilateral breast imaging, having a capability of four source wavelengths and 32 source-detector fibers for each breast, is described. The system records dynamic optical data simultaneously from both breasts, while verifying proper optical fiber contact with the tissue through implementation of automatic schemes for evaluating data integrity. Factors influencing system complexity and performance are discussed, and experimental measurements are provided to demonstrate the repeatability of the instrumentation. Considerations in experimental design are presented, as well as techniques for avoiding undesirable measurement artifacts, given the high sensitivity and dynamic range (1:10(9)) of the system. We present exemplary clinical results comparing the measured physiologic response of a healthy individual and of a subject with breast cancer to a Valsalva maneuver. PMID- 15835361 TI - Improvement of transcutaneous fluorescent images with a depth-dependent point spread function. AB - The point-spread function (PSF) for transcutaneous fluorescent imaging was obtained as an analytical solution in a closed form. It is applicable to cases in which the optical property of the image-blurring turbid medium is considered to be fairly homogeneous. We proposed a technique to improve a transcutaneous image by using depth-dependent PSF. Contrast of the fluorescent image was improved for depths of 1-15 mm in a scattering medium (micro(s)' = 1/mm). The visible depth was more than doubled with this technique. An experiment with a rat demonstrated considerable improvement of a transcutaneous image of the cerebral vein at a specified depth. The spread image of the heart was reduced to the correct size by use of the PSF with the actual depth of the heart. PMID- 15835362 TI - Adjustment of guidelines for exposure of the eye to optical radiation from ocular instruments: statement from a task group of the International Commission on Non Ionizing Radiation Protection (ICNIRP). AB - A variety of optical and electro-optical instruments are used for both diagnostic and therapeutic applications to the human eye. These generally expose ocular structures to either coherent or incoherent optical radiation (ultraviolet, visible, or infrared radiation) under unique conditions. We convert both laser and incoherent exposure guidelines derived for normal exposure conditions to the application of ophthalmic sources. PMID- 15835363 TI - Magnetic-resonance-imaging-coupled broadband near-infrared tomography system for small animal brain studies. AB - A novel magnetic-resonance-coupled broadband near-infrared (NIR) tomography system for small animal brain studies is described. Several features of the image formation approach are new in NIR tomography and represent major advances in the path to recovering high-resolution hemoglobin and oxygen saturation images of tissue. The NIR data were broadband and continuous wave and were used along with a second-derivative-based estimation of the path length from water absorption. The path length estimation from water was then used along with the attenuation spectrum to recover absorption and reduced scattering coefficient images at multiple wavelengths and then to recover images of total hemoglobin and oxygen saturation. Going beyond these basics of NIR tomography, software has been developed to allow inclusion of structures derived from MR imaging (MRI) for the external and internal tissue boundaries, thereby improving the accuracy and spatial resolution of the properties in each tissue type. The system has been validated in both tissue-simulating phantoms, with 10% accuracy observed, and in a rat cranium imaging experiment. The latter experiment used variation in inspired oxygen (FiO2) to vary the observed hemoglobin and oxygen saturation images. Quantitative agreement was observed between the changes in deoxyhemoglobin values derived from NIR and the changes predicted with blood oxygen-level-dependent (BOLD) MRI. This system represents the initial stage in what will likely be a larger role for NIR tomography, coupled to MRI, and illustrates that the technological challenges of using continuous-wave broadband data and inclusion of a priori structural information can be met with careful phantom studies. PMID- 15835364 TI - Multipass open-path Fourier-transform infrared measurements for nonintrusive monitoring of gas turbine exhaust composition. AB - The detection limits for NO and NO2 in turbine exhausts by nonintrusive monitoring have to be improved. Multipass mode Fourier-transform infrared (FTIR) absorption spectrometry and use of a White mirror system were found from a sensitivity study with spectra simulations in the mid-infrared to be essential for the retrieval of NO2 abundances. A new White mirror system with a parallel infrared beam was developed and tested successfully with a commercial FTIR spectrometer in different turbine test beds. The minimum detection limits for a typical turbine plume of 50 cm in diameter are approximately 6 parts per million (ppm) for NO and 9 ppm for NO2 (as well 100 ppm for CO2 and 4 ppm for CO). PMID- 15835365 TI - Diffraction-induced coherence levels. AB - We examined the influence of complex diffraction effects on low-coherence fringes created for high-aspect depth-to-width ratio structures called trenches. The coherence function was analyzed for these micrometer-wide trenches and was registered with a white-light interference microscope. For some types of surface structure we observed that additional low-coherence fringes that do not correspond directly to the surface topology are formed near the sharp edges of the structures. These additional coherence fringes were studied by rigorous numerical evaluations of vector diffractions, and these simulated interference fields were then compared with experimental results that were obtained with a white-light interference microscope. PMID- 15835366 TI - In vivo absorption spectroscopy of tumor sensitizers with femtosecond white light. AB - A system based on a femtosecond white-light continuum and a streak camera was used for recordings of the in vivo absorption spectra of the tumor-seeking agent disulphonated aluminum phthalocyanine. Measurements for different drug doses were performed on tumor tissue (muscle-implanted adenocarcinoma) and normal muscle tissue in rats. It was found that the shape of the spectrum is tissue dependent. The peak of the absorption spectrum is blueshifted in tumor tissue as compared with the muscle. Thus the contrast in the drug-related absorption can be altered by up to a factor of 2 from the primary drug molecular-concentration contrast between normal muscle and tumor by the proper selection of the illumination wavelength. PMID- 15835367 TI - Computerized case management: what technology can do for you. PMID- 15835368 TI - Ensure your system meets current and future needs. PMID- 15835370 TI - Holistic wound care yields better healing rates. PMID- 15835369 TI - Access program reduces inappropriate admissions. PMID- 15835371 TI - Is your falls prevention program getting results? PMID- 15835372 TI - CMs coordinate care for patient at end of life. PMID- 15835373 TI - What is results-driven case management? PMID- 15835375 TI - Decline of renal function is associated with proteinuria and systolic blood pressure in the morning in diabetic nephropathy. AB - The aim of this study was to investigate a significance of increased proteinuria in the morning and the effects of antihypertensive treatment on proteinuria and arterial blood pressure in the progression of chronic renal insufficiency in type 2 diabetic patients with hypertension and nephropathy. In three 24-hr urine samples and blood pressure monitoring, separated into a night-and daytime and spot urine in the morning, variation in protein-creatinine ratio (g/g) and blood pressure were assessed in 24 (58 +/- 3 years old; M/F: 17/7) diabetic patients with hypertension and nephropathy. Furthermore, the effects of antihypertensive therapy of combinations of angiotensin converting enzyme (ACE) inhibitor, calcium antagonists, diuretics, and alpha1 blocker were evaluated in 3 years. Home blood pressure measurement was carried out every month and 24-hr urine was collected every 2 months. The baseline urine excretion of protein-creatinine ratio and blood pressure were (1.22 +/- 0.13 g/g creatinine: 154/96 +/- 6/5 mmHg) in daytime and (1.39 +/- 0.13: 168/88 +/- 15/7) in the morning. At the end of the study, significant associations among a decline of 24-hr creatinine clearance and both of the urine excretion of protein-creatinine ratio (r = 0.47, p < .01) and the levels of systolic blood pressure (r = 0.46, p < .01) and between the levels of systolic blood pressure and the urine excretion of protein-creatinine ratio in the morning (r = 0.57, p < .001) were demonstrated. However, there were no significant associations among other variables. Analysis of patients who had systolic blood pressure in the morning less than 140 mmHg revealed that 65% of these patients received doxazosin-averaged doses of 4.8 +/- 1.5 mg daily. The levels of both blood pressure and proteinuria-creatinine ratio in the morning mainly associate with progression of renal function in diabetic patients with hypertension and nephropathy. PMID- 15835374 TI - Chronobiologically explored effects of Telmisartan. AB - Effects of Micardis (Telmisartan), alone or with low-dose aspirin, on blood pressure and other cardiovascular endpoints are examined in 20 patients with MESOR-hypertension in a crossover, double-blind, randomized study consisting of three stages, each lasting 7 days: I-placebo, II-Micardis, and III-Micardis with low-dose aspirin. Treatment was administered each day at a different circadian stage, upon awakening, and 3, 6, 9, 12, 15 and 18 hr after awakening. During each stage, the following variables were measured at 3-hr intervals during waking: systolic and diastolic blood pressure, heart rate, ejection fraction, intrarenal resistive index, acceleration time, and serum creatinine. Each data series was analyzed by single cosinor. Results were summarized by population-mean least squares spectra. At matched treatment times, the MESOR and circadian amplitude of each variable were compared among the three treatments by paired t-tests. A prominent circadian rhythm characterizes all variables. Micardis was associated not only with a lowering of blood pressure, but also with a reduction of the circadian blood pressure amplitude. The ejection fraction was increased, and the resistive index and acceleration time were decreased, the effect being more pronounced when low-dose aspirin was added to Micardis. Any circadian-stage dependent effect of Micardis, with or without low-dose aspirin, will require monitoring over spans longer than a single day for a given treatment administration time. PMID- 15835377 TI - Fractal and complexity measures of heart rate variability. AB - Heart rate variability has been analyzed conventionally with time and frequency domain methods, which measure the overall magnitude of RR interval fluctuations around its mean value or the magnitude of fluctuations in some predetermined frequencies. Analysis of heart rate dynamics by methods based on chaos theory and nonlinear system theory has gained recent interest. This interest is based on observations suggesting that the mechanisms involved in cardiovascular regulation likely interact with each other in a nonlinear way. Furthermore, recent observational studies suggest that some indexes describing nonlinear heart rate dynamics, such as fractal scaling exponents, may provide more powerful prognostic information than the traditional heart rate variability indexes. In particular, the short-term fractal scaling exponent measured by the detrended fluctuation analysis method has predicted fatal cardiovascular events in various populations. Approximate entropy, a nonlinear index of heart rate dynamics, that describes the complexity of RR interval behavior, has provided information on the vulnerability to atrial fibrillation. Many other nonlinear indexes, e.g., Lyapunov exponent and correlation dimensions, also give information on the characteristics of heart rate dynamics, but their clinical utility is not well established. Although concepts of chaos theory, fractal mathematics, and complexity measures of heart rate behavior in relation to cardiovascular physiology or various cardiovascular events are still far away from clinical medicine, they are a fruitful area for future research to expand our knowledge concerning the behavior of cardiovascular oscillations in normal healthy conditions as well as in disease states. PMID- 15835376 TI - Analysis of factors that affect short-term blood pressure variability in patients with chronic renal failure. AB - Recent reports suggest the relationship of short-term blood pressure (BP) variability to cardiovascular target organ damage. In this study, short-term BP variability was assessed as the standard deviation of daytime and nighttime BP in 36 hospitalized patients with chronic renal failure (CRF) who underwent ambulatory BP monitoring. Positive correlations were observed between body mass index (BMI) and daytime systolic and diastolic BP variability, BMI and nighttime diastolic BP variability, cholesterol and daytime systolic BP variability, cholesterol and nighttime systolic and diastolic BP variability, nocturnal decline in BP and nighttime diastolic BP variability, and plasma concentration of norepinephrine (p-NE) and nighttime systolic BP variability. In multivariate linear regression analyses, BMI showed the strongest association with daytime and nighttime diastolic BP variability (p < .005 and p < .05). On the other hand, cholesterol and p-NE were the primary determinants of daytime and nighttime systolic BP variability, respectively (p < .01 and p < .0005). Interestingly, CRF patients with ischemic heart disease (IHD) had significantly increased daytime systolic and diastolic BP variability and nighttime systolic BP variability (p < .05 or less). Furthermore, logistic regression analysis demonstrated that nighttime systolic BP variability was an independent risk factor of IHD in patients with CRF (odds ratio 1.50 [95% confidence interval 1.01 to 2.25]; p < .05). Taken together, short-term BP variability is suggested to be affected by BMI, cholesterol, and p-NE in CRF patients. Furthermore, sympathetic nerve overactivity may be involved in cardiovascular complications in CRF patients through the increase in nighttime systolic BP variability. PMID- 15835378 TI - Circadian expression of plasminogen activator inhibitor-1 in angiotensin II type 1a receptor knockout mice. AB - Both the peripheral biological clock and the renin-angiotensin system regulate mRNA expression of plasminogen activator inhibitor-1 (PAI-1). Our objective was to determine whether angiotensin II (Ang II) type 1 (AT1) receptor-mediated signaling contributes to the development of circadian expression of PAI-1 and clock genes in the heart, aorta, liver, and kidney. We sacrificed AT1a receptor knockout (AT1a-KO) and wild-type (WT) 12 week-old mice every 4 hr. We examined mRNA expression for PAI-1 and clock genes (Per2, Bmal1, and Clock) in heart, aorta, liver, and kidney by using the quantitative reverse transcription polymerase chain reaction. PAI-1 mRNA showed circadian oscillation with a peak occurring during the light phase in the heart, liver, aorta, and kidney of WT mice. Peak expression of PAI-1 in the liver and aorta was decreased in AT1a-KO mice. On the other hand, cardiac PAI-1 expression in AT1a-KO mice was reduced in the dark phase, during which time its expression level was low. There were no significant differences between WT and AT1a-KO mice in renal PAI-1 expression. Clock genes oscillated synchronously in WT and AT1a-KO mice, and there were no significant differences between the WT and the AT1a-KO mice in their expression. Plasma angiotensin II showed little oscillation in the WT mice. We conclude that AT1a receptor-mediated Ang II signaling modulates the circadian expression of PAI 1 in an organ-specific manner. The effect of the renin-angiotensin system on PAI 1 expression appears to be independent of peripheral clock gene expression. PMID- 15835379 TI - Implication of base heart rate in autonomic nervous function, blood pressure and health-related QOL. AB - Increased resting heart rate (HR) and increased sympathetic nervous activity are independent risk factors for cardiovascular disease. Recently, base heart rate (HRo: minimum stable HR during sleep) has been reported to relate to cardiac stroke volume and age. However, little is known about the relevance of HRo. The aim of our study was to evaluate how HRo is associated with HR variability (HRV), blood pressure and health-related quality of life (HRQOL) in healthy subjects. A total of 139 volunteers participated in this study that measured 24-hr HR, HRV, and blood pressure. HRo was estimated from the trendgram and the histogram of HR during the nighttime (sleep) period, and calculated as the 1% lowest value of its integral. HRQOL was assessed by Medical Outcome Study Short-Forum 36-Item Health Survey. Sympathetic nervous activity (ratio of low frequency to high frequency component: LF/HF) and parasympathetic nervous activity (high frequency component: HF) were calculated by ECG monitoring. HRo was positively correlated with 24-hr LF/HF and nighttime LF/HF. HRo was negatively correlated with 24-hr HF and nighttime HF. Moreover, HRo was positively correlated with the scores of social functioning and role-physical. Using multivariate analysis, HRo is related to LF/HF, body mass index, and the score of social functioning (HRQOL). HRo may be a useful indicator for assessing sympathetic nervous activity and HRQOL in normotensive healthy subjects. PMID- 15835380 TI - ACE inhibitors and chronotherapy. AB - Chronotherapy can improve the effectiveness and reduce the adverse reactions of drugs and actually is used for several conditions including cardiovascular diseases. Although angiotensin converting enzyme (ACE) inhibitors are available for the therapy of patients with hypertension and/or heart failure, these agents have some characteristic adverse effects such as angioedema and dry cough. It has been reported that the dosing of ACE inhibitor at an inactive period has a better protective effect against cardiac hypertrophy in hypertensive rats, and changing dosing time from morning to evening reduces the severity and frequency of the drug-induced dry cough of hypertensive patients treated in the morning. Thus, the dosing of ACE inhibitors in the inactive span is more effective and safe. Dosing in the evening may be an alternative for hypertensives with dry cough with a morning dose of ACE inhibitors, if one ascertains that no circadian hyperamplitude tension is induced by the evening dose of this or any other antihypertensive drug. PMID- 15835381 TI - Quantitative study of circadian variations of ambulatory blood pressure in Chinese healthy, hypertensive, and diabetes subjects. AB - Ambulatory blood pressure monitoring (ABPM) recorded abundant data of BP and heart rate (HR) variations with even more derived parameters for evaluation of BP. Using our ABP database system established recently, we studied quantitatively the data of 24-hr ABP in Chinese. First, 155 Chinese were divided into three groups: 50 healthy subjects (C) of 20 men and 30 women, aged 60.0 +/- 10.3 (SD) years; 58 hypertensive patients (H, mild or moderate hypertension) of 33 men and 25 women, aged 59.4 +/- 8.0 years; 47 diabetes patients (D, type 2 diabetes, all were normotensive and with no insulin treatment) with 28 men and 19 women, aged 61.0 +/- 8.5 years. Then 24-hr ABP was monitored by TM-2421 Monitor and data were analyzed by ABP database, cosinor method, and conventional statistics. Our results were 4-fold: 1) systolic BP (SBP), diastolic BP (DBP), HR, rate-pressure product (HR x SBP) showed circadian variations, and significant circadian rhythms were confirmed by cosinor method in all groups. MESOR (midline estimate statistic of rhythm) differed significantly among three groups (H had the highest and C had the lowest values); 2) BP means (SBP, DBP, pulse pressure [PP], and HR x SBP) and BP loads (SBP, DBP, and PP) showed significant differences among the groups (H and D had higher values than that of C); 3) there were no significant differences of BP variability (BPV) of SBP, DBP, and PP among the groups; 4) areas under curve of BP (SBP, DBP, and PP) in H were significantly higher than in C and there was no significant difference between H and D. We concluded that ABPM can offer abundant information on BP evaluation by its direct recording data and derived parameters. The computerized way of treating the large numbers of ABPM values supplies a useful tool in evaluation of BP. Our results suggest that clinically normotensive diabetes patients had some pathological alterations in their BP systems. PMID- 15835382 TI - Overweight influence on circadian variations of ambulatory blood pressure in Chinese adolescents. AB - Childhood obesity and its consequences have been the subject of intense interest in recent years. In this study we examined the influence of overweight on circadian variations of ambulatory blood pressure (ABP) in Chinese adolescents. First, 24-hr ABP monitoring was performed in 252 adolescents divided into two groups with equivalent sex, age, and body height (49 girls and 77 boys in each group): controls (normal weight) were aged 13.68 +/- 1.21 years, height 165.37 +/ 9.45 cm, body mass index (BMI) 18.82 +/- 2.3; overweights (BMI > or = 24) were aged 13.71 +/- 1.23 years, height 165.75 +/- 9.47 cm, BMI 27.70 +/- 3.1. ABP recordings were treated by ABP database system and analyzed by cosinor method and conventional statistics methods. The circadian variations of ABP in adolescent patterned as "dipper" and circadian rhythmicity of ABP variations were confirmed by cosinor analysis in most adolescents of both groups. Significant statistical differences were found for rhythm parameters: the MESOR (midline estimate statistic of rhythm), peak, trough (the maximum and minimum values derived from the composed curves, respectively), and amplitude values between control and overweight groups. Significant higher values also were seen in the overweight group for most of ABP parameters (p < .01), such as, BP means (SBP, DBP, MAP: mean arterial pressure, or PP: pulse pressure), BP variability, BP loads and rate pressure product (HR x SBP). Our results have shown that overweight influenced significantly on ABP and parameters derived from ABP recordings in Chinese adolescents, which suggests an increasing risk of cardiovascular diseases in overweight adolescents. PMID- 15835383 TI - Development of wristwatch-type heart rate recorder with acceleration-pickup sensor and its application. AB - We have developed a wristwatch-type heart rate (HR) recorder (WT-HR tachometer) with an acceleration-pickup sensor, which can record HR and body activity (ACT) minute by minute for a long time (> 120 h). To study its application, we compared these measurements with 24-h blood pressure (BP) and HR variability, sympathovagal balance, and resting energy expenditure (REE). The WT-HR tachometer incorporates a Seiko-blue sensor to measure HR at the wrist and an acceleration pickup sensor to measure body ACT every 4 sec. Fifty healthy university students underwent measurement of HR and ACT by the WT-HR tachometer for 24 h. Then we calculated the minimum HR and mean HR during waking time and derived deltaHR. On the same day, 24-hr BP and LF/HF (low frequency component/high frequency component of ECG-RR interval power spectrum) were measured with a multibiomedical recorder, and REE was measured with a portable calorimeter. The WT-HR tachometer accurately measured HR, compared with the measurements by Holter-ECG. There was a significant correlation between deltaHR and deltaSBP. There also was a significant correlation between deltaHR and deltaLF/HF, (waking time = waking time without body movement) and a significant correlation between HR and REE. These results suggest that ACT and deltaHR are useful for evaluating mental and physical activity of daily life. The WH-HR tachometer will be useful for assessment of mental and physical activity in daily life. PMID- 15835384 TI - Influence of repeated measurement on one occasion, on successive days, and on workdays on home blood pressure values. AB - Few reports provide clear guidelines on how home blood pressure (HBP) should be measured in practice. In this study, we evaluated the influence of repeated HBP measurements on one occasion, and the difference between the 1st and 2nd day and between workdays and nonworkdays. The subjects (468 male, 232 female; mean age 41 years) were recruited from one company. HBP was measured with a semiautomatic device (Omron HEM-759P). Subjects were instructed to perform triplicate morning (m) and evening (e) measurements on 7 consecutive days. HBP tended to decrease during repeated measurements: systolic blood pressure (SBP) was significantly higher the 1st time than the 2nd time and 3rd time. There was no difference in diastolic blood pressure (DBP) between the 1st and 2nd time, but the value the 3rd time was significantly lower than the 1st and 2nd time. Both mHBP and eHBP on the 1st day were significantly higher than those on the 2nd day. mHBP was higher on the 1st workday than on nonworkdays, but the difference was less than 1 mmHg and there was no significant difference. Since there were significant differences in HBP during repeated measurements and between the 1st and 2nd day, which value to adopt as HBP needs to be discussed. Whether HBP was measured on a workday or a nonworkday seemed to have little influence on the HBP values obtained. PMID- 15835385 TI - Severe emotional hypertension compatible with 95-year lifespan and little end organ damage: 24-year record. AB - We report on an interesting case of longevity in an elderly Japanese woman whose blood pressure (BP) continued extremely high from her first recording at the age of 38 years to her first hospitalization at the age of 81. BP recordings taken by her physician indicated mostly severe or occasionally mild hypertension (HT): between 260/130 and 140/76 mmHg. No antihypertensive drugs were taken during the 25 year span between ages 56 and 80. After her physician died, when she was 80, she changed clinicians and was given several kinds of antihypertensive drugs. She began to feel faintness, dizziness, and severe fatigue. At the time of the first hospitalization, no remarkable cerebral or cardiac abnormalities were observed, despite her BP as high as 210/110 mmHg. BP as measured by nurses or the physician in charge were always above 160/80 mmHg. After discharge, she was asked to measure BP by herself at home. These readings were 140-150/70-80 mmHg on the average, indicating a rare case of long-term emotional blood pressure response. The patient died not of a cerebrocardiovascular accident, but of acute renal failure at 95 years of age. Even though her recorded BP was extremely high when measured by medical staff members and still far above the current conventional desired limit of 120 mmHg systolic (S) BP or the earlier limit of 140 mmHg SBP, it was actually acceptable for her retrospectively, insofar as she lived to age 95. Although antihypertensive drug therapy may be helpful in some cases, it may not be necessary in others. Intensive drug therapy may even be harmful for misdiagnosed emotionally HT patients particularly those misdiagnosed with refractory hypertension, when the response to health care professionals may be emotional. PMID- 15835386 TI - Weekly variation of home and ambulatory blood pressure and relation between arterial stiffness and blood pressure measurements in community-dwelling hypertensives. AB - Although blood pressure (BP) is a major determinant of pulse wave velocity (PWV), some treatments have independent effects on BP and arterial stiffness. Although both ambulatory BP (ABP) and self-measured BP at home (HBP) have become important measures for the diagnosis and management of hypertension, single day recordings may be insufficient for a proper diagnosis of hypertension or the evaluation of treatment efficacy. To evaluate weekly variations in BP using 7-day HBP and 7-day ABP monitoring and to determine the relation between arterial stiffness and BP measurements in community-dwelling patients with hypertension. We enrolled 68 community-dwelling hypertensive subjects in this study. Significant weekly variations in systolic blood pressure (SBP) and diastolic blood pressure (DBP) were found in the awake ABP data (p < .01, respectively), while no significant weekly variations in the asleep ABP or the morning and evening HBP data were observed. In untreated subjects, significant correlations were obtained between the brachial-ankle PWV and the average awake SBP, the average asleep SBP and the average SBP measured by HBP in the evening. In treated subjects, only the average SBP measured by HBP in the morning was significantly correlated with the baPWV. Differences in the weekly variations in BP were observed between HBP and ABP monitoring. In addition, the morning systolic HBP was not correlated with arterial stiffness in untreated subjects with hypertension but was correlated in treated subjects. Relations between the morning HBP and arterial stiffness might be attributed to morning surges in BP and/or trough levels of antihypertensive drugs. PMID- 15835387 TI - Circadian dynamics of heart rate and physical activity in patients with heart failure. AB - The present study was designed to develop a method to continuously measure Holter electrocardiogram (ECG) and physical activity in terms of metabolic costs to examine circadian dynamics of RR intervals and physical activity in patients with heart failure. A total of 7 healthy subjects and 3 heart failure patients performed cardiopulmonary exercise test using four-stage graded treadmill walking at 0% grade to examine whether the acceleration signals in the vertical direction could reflect actual body energy expenditure during physical activity. Then, using this new method, 24-hr monitorings of ECG and physical activity were performed in 24 inpatients with heart failure while they were allowed to walk around freely. Our results showed the integral of rectified acceleration signals was closely correlated with actual metabolic cost in all subjects. Instantaneous changes in heart rate were quite concordant with physical activity. As compared with the asymptomatic patients (n = 12), the symptomatic patients (n = 12) had lower energy expenditure during 8-hr daytime periods but higher mean heart rate. Furthermore, a more prominent ultradian rhythm of circadian changes in heart rate and physical activity was found in 50% of all subjects studied. The simultaneous analysis of Holter ECG and physical activity as the same time series revealed that in patients with heart failure, sympathovagal balance shifted toward sympathotonic conditions and their physical activity could become subject to intrinsic ultradian dynamics of body's homeostasis. PMID- 15835388 TI - Correlation of heart rate turbulence with sympathovagal balance in patients with acute myocardial infarction. AB - To examine the relationship among heart rate turbulence parameters, arterial baroreflex sensitivity, and cardiac sympathetic nerve activity, 15 patients with acute myocardial infarction, presenting with sinus rhythm and > or = 3 ventricular premature beats/24 hr were studied at least 2 weeks after acute myocardial infarction. Turbulence onset (TO) and turbulence slope (TS) were averaged from 3 respective ventricular premature beats. Early heart-to mediastinum ratio (H/M), delayed H/M, and washout rate were calculated from iodine-123-metaiodobenzylguanidine (123I MIBG) scintigraphy. Arterial baroreflex sensitivity was calculated by phenyrephrine method. Arterial baroreflex sensitivity correlated significantly with TO (r = - 0.75, p < .01) and TS (r = 0.53, p < .05). TO had no correlations with early H/M, delayed H/M, and washout rate. There were no significant correlations between TS and early H/M. However, TS had significant correlation with delayed H/M (r = 0.74, p < .01) and washout rate (r = -0.71, p < .01). Thus, heart rate turbulence of TO and TS parameters depend on sympathovagal balance. PMID- 15835389 TI - Successful treatment of obstructive sleep apnea syndrome improves autonomic nervous system dysfunction. AB - Autonomic nervous system (ANS) dysfunction may be implicated in the subsequent development of cardiovascular disease in patients with obstructive sleep apnea syndrome (OSAS). To confirm the relation between OSAS and ANS dysfunction, we prospectively investigated ANS function in 7 patients with moderate or severe OSAS; 7 healthy age-matched volunteers were for control. We also studied ANS function before and after treatment in the patients with OSAS to evaluate the effect of OSAS treatment on ANS dysfunction. The body mass index of patients with OSAS was 32.2 (27.4-45) (median [range]) kg/m2. The patients were treated by nasal continuous positive airway pressure (n = 5) or uvulopalatopharyngoplasty (n = 2). The apnea/hypopnea index decreased markedly from 42.1 (30.6-77.2) events/hr of sleep before treatment to 2.3 (1.4-3.8) after treatment. To evaluate ANS function, the coefficient of variation of the RR interval (CV-RR) and corrected QT (QTc) interval on the electrocardiogram at rest and the heart rate (HR) responses to blood pressure (BP) changes during the Valsalva maneuver were studied. Baseline HR of OSAS patients was significantly higher than that of the control subjects (p < .05). The Valsalva ratio (VR), baroreflex sensitivity (BRS), and CV-RR values in patients with OSAS were significantly lower than those of the control subjects (all, p < .005). However, there were no significant differences in systolic and diastolic BP or QTc intervals. After treatment, VR, BRS, and CV-RR values increased significantly compared with those before treatment in patients with OSAS (all, p < .05). There were no significant differences in systolic and diastolic BP, HR, or QTc intervals measured before and after treatment. These results suggest that impaired ANS function is present in patients with OSAS and can be improved by successful treatment of OSAS. PMID- 15835390 TI - Effects of hydroxyfasudil administered to the nucleus tractus solitarii on blood pressure and heart rate in spontaneously hypertensive rats. AB - Previously, we reported that the inhibition of Rho-kinase by a microinjection of Y-27632 or the transfection of dominant-negative Rho-kinase into cells of the nucleus tractus solitarii (NTS) reduces blood pressure, heart rate, and sympathetic nerve activity. In the present study, we examined the effects of another Rho-kinase inhibitor, hydroxyfasudil, on blood pressure and heart rate in anesthetized rats. The results were compared between normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). The microinjection of hydroxyfasudil was performed unilaterally or bilaterally into the NTS of WKY rats and SHR. A unilateral microinjection of hydroxyfasudil elicited depressor and bradycardic responses in SHR but not in WKY rats. A bilateral microinjection of hydroxyfasudil elicited depressor and bradycardic responses in both SHR and WKY rats. However, the magnitude of the decrease in these variables was greater in SHR than in WKY rats. The expression levels of RhoA in the membrane fraction and phosphorylated ERM family (ezrin, radixin, and moesin) in the NTS were greater in SHR than in WKY rats. These results suggest that the microinjection of hydroxyfasudil into the NTS causes cardiovascular responses similar to those caused by Y-27632 and that these responses are probably mediated by the inhibition of Rho-kinase. PMID- 15835391 TI - Ring-shaped pulse oximeter and its application: measurement of SpO2 and blood pressure during sleep and during flight. AB - Respiratory and cardiovascular functions show circadian and day-to-day changes. We have developed a wireless ring-shaped pulse oximeter in collaboration with MC Medical Inc. and Advanced Medical Inc. We investigated the accuracy of this pulse oximeter and its application in daily life. Percutaneous arterial oxygen saturation (SpO2) of 47 volunteers was measured simultaneously with the ring shaped pulse oximeter and a standard pulse oximeter. A total of 103 volunteers underwent measurement of SpO2 for 24 hr, and 11 healthy volunteers underwent measurement of SpO2 and blood pressure (BP) during flight. SpO2 and heart rate (HR) were measured and recorded every 20 sec, cabin barometric pressure and cabin oxygen concentration equivalent to sea level were measured minute-to-minute, and BP was measured every 3 min with a portable BP recorder during each flight. The SpO2 values measured with the ring-shaped pulse oximeter were similar to those measured with the standard method. The mean SpO2 during sleep was significantly lower in the group with high-normal BP or mild hypertension than in the group with normal BP. During flight, the mean change in SpO2 was -2.4 +/- 1.7% during nose-up flight, and 2.1 +/- 2.6% during nose-down flight. There was a significant correlation between change in SpO2 and change in systolic BP during nose-up flight. The wireless ring-shaped pulse oximeter was useful for investigating changes in SpO2 and its effect on BP in daily life during sleep and during air travel. PMID- 15835392 TI - Evaluation of the severity of dementia using expectation control function. AB - The results of an animal experiment without anesthesia suggested the presence of expectation control in which sympathetic nerve tone increases at rest immediately before behaviors to prepare for the subsequent behavior. We investigated whether such function is present in humans in their daily living and whether patients with dementia could be differentiated from healthy adults based on this finding. Using a Holter electrocardiograph with an acceleration sensor, electrocardiograms (ECG) and behavioral states were recorded. The total recorded ECG was divided by 20 sec, and high (HF, 0. 15-0.4 Hz) and low (LF, 0.03-0.15 Hz) frequency components were extracted by frequency analysis of the RR interval. We used the HF component, L/H ratio, and RR interval as the index grade of parasympathetic/sympathetic nerve tone. The mean value of each parameter was calculated and compared between the time zone during which resting persisted for 1 min or more (3 intervals) and that during which behaviors occurred immediately after a 1-min rest., In 32 patients with dementia, Holter ECG and behaviors were serially recorded, and the time zone during which intentional behaviors occurred was detected. RR interval and HF component decreased and L/H increased. Increasing in sympathetic nerve tone was observed at rest condition immediately before intentional behaviors, suggesting a presence of expectation control function. The expectation control mechanism will present in humans and involved in this function by severity of dementia. This parameter may be useful for objectively evaluating the severity of dementia. PMID- 15835393 TI - Personality and mortality from ischemic heart disease and stroke. AB - BACKGROUND: The role of personality in the causation of circulatory diseases has been controversial. METHODS: From June through August 1990, 41,442 residents of Miyagi Prefecture in northern Japan completed the Japanese version of the short form Eysenck Personality Questionnaire-Revised and another questionnaire on various health habits. During 11 years of follow-up until March 31, 2001, we identified 90 deaths from ischemic heart disease (IHD) and 131 deaths from stroke. We used Cox regression to estimate the relative risk of IHD and stroke according to the three levels of four personality subscales (extraversion, neuroticism, psychoticism, and lie), with adjustment for sex, age, and other potentially confounding variables. RESULTS: Multivariate relative risks of IHD for the highest verses the lowest level of personality subscales were 0.7 for extraversion, 1.1 for neuroticism, 1.3 for psychoticism, and 0.8 for lie. Multivariate relative risks of stroke for the highest verses the lowest level of personality subscales were 1.0 for extraversion, 0.9 for neuroticism, 1.2 for psychoticism, and 1.2 for lie. CONCLUSIONS: This prospective study does not support the hypothesis that personality is a risk factor for mortality from IHD and stroke. PMID- 15835395 TI - [Opening speech from the 99th meeting of the Anatomy Society in Vienna, April 2 5, 2005]. PMID- 15835394 TI - Cardiovascular tissues contain independent circadian clocks. AB - Acute cardiovascular events exhibit a circadian rhythm in the frequency of occurrence. The mechanisms underlying these phenomena are not yet fully understood, but they may be due to rhythmicity inherent in the cardiovascular system. We have begun to characterize rhythmicity of the clock gene mPer1 in the rat cardiovascular system. Luciferase activity driven by the mPer1 gene promoter is rhythmic in vitro in heart tissue explants and a wide variety of veins and arteries cultured from the transgenic Per1-luc rat. The tissues showed between 3 and 12 circadian cycles of gene expression in vitro before damping. Whereas peak per1-driven bioluminescence consistently occurred during the late night in the heart and all arteries sampled, the phases of the rhythms in veins varied significantly by anatomical location. Varying the time of the culture procedure relative to the donor animal's light:dark cycle revealed that, unlike some other rat tissues such as liver, the phases of in vitro rhythms of arteries, veins, and heart explants were affected by culture time. However, phase relationships among tissues were consistent across culture times; this suggests diversity in circadian regulation among components of the cardiovascular system. PMID- 15835396 TI - The collagenic structure of human digital skin seen by scanning electron microscopy after Ohtani maceration technique. AB - We performed a morphological scanning electron microscope (SEM) study to describe the fine structure and disposition of collagenous tissue in the human toe. After therapeutic amputation of a human right leg, we applied the Othani maceration technique to the skin of three toes surgically explanted from the foot. We distinguished eight cutaneous regions and focused on some specialized collagenous structures differing in the thickness of the skin. The eight areas investigated were: the dorsal skin, the eponychium, the perionychium, the hyponychium, the region under the visible nail, the nail root, the plantar skin and finally the toe tip. Each of these areas is characterized by a distinctive collagenous surface disposition, with some peculiar features mostly related to dermal papillae. At high magnification, we observed the spatial arrangement of the collagen fibers constituting the top of the dermal papillae that represents the attachment site of the proliferative basal layer of the epidermis. We also noted an impressive density of collagen fibers throughout the thickness of the dermal layer, organized in specialized structures and constituting the skeleton of dermal thermoreceptorial corpuscles or sweat glands. A combination of SEM and Ohtani technique disclosed the three-dimensional architecture of the collagenous matrix of tarsal skin under physiologic conditions, giving a detailed description of the most reactive tissue during pathologic processes. PMID- 15835397 TI - Features of the peritoneal covering of the lesser pelvis with special reference to stomata regions. AB - Occasional reports describe various aspects of the fine morphology of the pelvic peritoneum, but its complete organ characteristics remain undefined. The peritoneal covering of the urinary bladder, rectum, uterus, uterine tube, ovary, broad ligament (BL) and testis in Wistar rats was examined by means of transmission and scanning electron microscopy (TEM, SEM). Unusually complicated relief and stomata between the cubic mesothelial cells characterized the surface of the BL. Deep, parallel furrows separated the wide longitudinal folds over the entire length of the uterine tube. The uterus and the ovary formed less numerous, shallow or extremely deep crypt-like invaginations, as well as serous villus-like or papilla-like evaginations. The flat cells were the predominant cell type over the BL, while the cubic mesothelium was the basic covering of the organs. Most of the cubic cells were located in the invagination of the submesothelial layer (SML). Such cells formed an almost smooth surface over the urinary bladder or formed larger areas of the rectum and the testis surfaces. Numerous microvilli, ciliae, round evaginations and complex lamellar bodies characterized their apical plasmalemma. In conclusion, the mesothelial heterogeneity is a stable feature of the lesser pelvis peritoneum, confirmed by TEM and SEM. The cubic mesothelium characterizes the organ peritoneum, while the BL plays the role of the parietal sheet, involving lymphatic units in the SML. The different types of contacts between the mesothelio-endothelial cells, large lymphatic vessels and occasional stomata are the usual components of the lymphatic units in norm, visible by TEM. Images of stomata, seen by SEM, demonstrate oval-shaped deep channel-like gaps surrounded by cubic mesothelium. The last data extend the evidence on stomata regions, which resemble the diaphragmatic ones. Clusters of cells (macrophages, mastocytes and Lymphocytes), small vessels (blood or lymphatic) and nerve fibers (unmyelinated and rare myelinated) form highly specialized complexes in the SML of the ovary, the uterus and the testis. PMID- 15835398 TI - Proteoglycans and collagen in the intervertebral disc of the rhesus monkey (Macaca mulatta). AB - The different varieties of supportive tissues in the intervertebral discs of the rhesus monkeys (Macaca mulatta) were investigated with regard to morphology of the collagen fibrils and distribution and localization of proteoglycans (PG). The annulus fibrosus and the nucleus pulposus of the intervertebral disc were structurally closely integrated into the cartilaginous endplates of the vertebral bodies. The collagen fibrils in the intervertebral disc fell into two categories; i.e. thick (70-110 nm) and thin (40-50 nm) ones. In the outer zone of the annulus fibrosus only thick fibrils occurred, while in the regions of the inner part of the annulus fibrosus, in the periphery of the nucleus pulposus and in the cartilaginous endplates both types of fibrils were found. PG were found free in the matrix and in great numbers in association with collagen fibrils in all regions of the intervertebral disc. They interconnected neighbouring fibrils and decorated the surface of the collagen fibrils in irregular orientation. Only the thick fibrils of the annulus fibrosus contained small regularly arranged intrafibrillar PG precipitates. The free precipitates were usually longer and thicker than the collagen-associated PG, in addition they could be branched. Since rhesus monkeys are closely related to humans, they can serve as model organisms and the findings presented may be of relevance to the understanding of the human intervertebral discs. PMID- 15835399 TI - An ultramicroscopic study on the distribution of Muller cell processes in the outer retinal layers of the zebrafish. AB - The zebrafish, Brachydanio rerio, is a good model for studying the development of various organs. We have assayed the distribution pattern of Muller cell processes in zebrafish retinas by electron microscopy. In the outer nuclear (ON) layer, multiple layers of Muller cell processes were present along both sides of the large pyramidal endings of the synaptic terminals. We found that the inner segments (ISs) of the zebrafish photoreceptors (PRs), including the cones, double cones and rods, were arranged in different planes, and that the Muller cell processes formed multilayered sheaths around virtually all PR compartments except their outer segments. Thus, Muller cell processes beyond the outer limiting membrane (OLM) are more easily observed in zebrafish retina than in the retinas of other species. To our knowledge, this is the first study to show the exact ultrastructural distribution of Muller cell processes around the OLM and the PR layer in zebrafish retina. PMID- 15835400 TI - The muscular organization of the stomach of capybara (Hydrochoerus hydrochaeris): an architectural view. AB - Twenty-two stomachs from adult capybaras were used in this study, and an acid digestion mesoscopic technique was pursued using different concentrations of nitric acid to observe the muscular organization of the stomach. The capybara's stomach possessed a muscular coat composed of four layers or strata: external longitudinal, external oblique, circular and internal oblique. Also, the cardiac and pyloric sphincter muscles were comprised of three or two different layers, respectively. Furthermore, the internal oblique fibres were observed extending from the cardiac portion of the stomach to the smaller curvature, where they participated in the formation of the Ansa cardiaca together with the external longitudinal fibres. This muscular architectural arrangement was compared to that in small rodents (rat, hamster, guinea pig), as well as in rabbits and pigs. In conclusion, the stomach of the capybara has a very particular, complex and defined muscular organization that differs from that in other rodents, or domestic animals, in particular, pigs. PMID- 15835401 TI - Prostaglandins of the E-series inhibit connective tissue proliferation in the liver wound of the rat. AB - The present study was undertaken to relate wound healing of an internal organ to prostaglandins of the E and F series. A small liver wound was induced by a galvanic cauter via the abdominal route under general anesthesia and prostaglandin E1, E2 and F2alpha were injected twice daily at a dose of 250 microg/kg. Proliferation of the connective tissue in the liver wound was estimated morphometrically 6 days after liver wound infliction. Levels of prostaglandins E2 and F2alpha were measured in the liver wound as well as in normal liver tissue from adjacent lobes using radioimmunoassay. The results show that exogenous prostaglandins of the E-series suppress connective tissue proliferation. Three minutes after the last prostaglandin E2 injection, high prostaglandin concentrations were measured both in the liver wound and in the liver tissue of the adjacent lobe. Prostaglandin F2alpha injections had no effect on wound healing. We believe that the rat thermic liver wound model can be used for different studies on wound healing mechanisms and that prostaglandins of the E-series are involved in wound healing in the specific time period studied. PMID- 15835403 TI - Analysis of thyroid gland microvascularization in rats induced by ingestion of potassium bromide: a scanning electron microscopy study. AB - The microvascularization of the thyroid gland in rats was studied in animals following ingestion of potassium bromide in various concentrations in drinking water (15, 50 or 100 mg/l) over a period of either 16 or 66 days. Accordingly, animals were divided into six study groups and one control group. Microvascularization was compared among the groups. The pattern of thyroidal follicle distribution within the blood vessels was evaluated by scanning electron microscopy. Animals that ingested potassium bromide presented an increased density and number of meshes in the capillary network of the follicle. The peripheral vessels showed flattening of the walls and in the central portion of the follicle, the distribution of the capillary network did not undergo any morphologic alteration. PMID- 15835402 TI - Morphological changes in experimental postischemic rat kidney. A pilot study. AB - The aim of our study was to investigate the changes in the early stages (at weeks 2 and 4) of experimental acute renal failure after short-time ischemia reperfusion (I/R) compared with the impact of Losartan. Twenty male Wistar rats were divided into three groups: sham-operated rats (2 weeks), I/R groups (2 and 4 weeks); I/R and Losartan-treated groups (2 and 4 weeks). I/R was produced in adult rats by clamping the left kidney renal artery and renal vein for 40 min. The angiotensin II receptor antagonist Losartan was added to the drinking water (40 mg/l), and treatment was started on the first day after the I/R. Body weight, systolic blood pressure (SBP) and 24 h urine amount was measured every week. Urine amount and SBP was higher in I/R groups compared to sham-operated rats. Early stage acute renal disease was characterized by focal segmental glomerulosclerosis (FSGS) and interstitial fibrosis (IF) at weeks 2 and 4 after I/R. In the Losartan group, 2 weeks after the surgery, FSGS, IF and mesangial cell proliferation was decreased, but at week 4 these parameters showed a tendency to increase. Marked changes take place in tubular epithelial cells, especially in I/R groups. Angiotensin II receptor blocker AT1RA Losartan in the small dose (40 mg/l) had no effect on hypertension and urine excretion in the experimental I/R model. A pilot study revealed that tubular basement membrane thickness is markedly increased after I/R. PMID- 15835404 TI - Immunohistochemical characterization of noncollagenous matrix molecules on the alveolar bone surface at the initial principal fiber attachment in rat molars. AB - This study was designed to immunodetect proteoglycans (PGs) and the noncollagenous glycoproteins, bone sialoprotein (BSP) and osteopontin (OPN) on developing alveolar bone surface in rat molars by the indirect immunoperoxidase method, and to discuss the roles of these molecules at the initial principal fiber (PF) attachment. To characterize PGs, antibodies against five species of glycosaminoglycans (GAGs), chondroitin-4-sulfate (C4S), chondroitin-6-sulfate (C6S), unsulfated chondroitin (C0S), dermatan sulfate (DS), and keratan sulfate (KS) were used. Maxillary alveolar bone facing the distal root of the second molar was examined in 20- and 25-day-old male Wistar rats. Routine histological staining was also used. A hematoxylin-stained, fibril-poor layer always appeared on the alveolar bone surface just prior to the initial PF organization. This layer was strongly immunoreactive for C4S, C0S, OPN, and BSP, and weakly for C6S, but not for DS and KS. Then the initial PFs were attached to this layer. When new bone containing Sharpey's fibers covered this layer, it remained as a hematoxylin stained, fibril-poor layer between Sharpey's fiber-containing and -lacking bone. The layer was consistently immunoreactive for OPN and BSP but had no immunoreactivity for GAGs. The results suggest that the accumulation of C4S-, C0S , and C6S-carrying PGs, and of BSP and OPN is a primary event at the initial PF attachment, and is involved in the adhesion of PFs and mineralization of the initial attachment layer. The BSP and OPN act to maintain the interface integrity between Sharpey's fiber-containing and Sharpey's fiber-lacking alveolar bone after the PF attachment is established. PMID- 15835405 TI - An uncommon form of the rare extensor carpi radialis accessorius. AB - Two accessory muscles were found in the lateral compartment of the forearm while dissecting a 92-year-old female cadaver. One of these originated from the extensor carpi radialis brevis, became tendinous and travelled between the two radial extensor tendons. It inserted independently into the second metacarpal bone, and may be regarded as an extensor carpi radialis intermedius. The other accessory muscle originated from the extensor carpi radialis longus, passed superficially over the parent tendon and inserted into the abductor pollicis brevis. This variation appeared to be a rare extensor carpi radialis accessorius, an additional muscle, which usually arises from below the extensor carpi radialis longus and inserts into the first metacarpal bone. Various forms of the accessorius have been described previously, including one that inserts into the abductor pollicis brevis. The tendon of the accessory muscle described in this study passed through its own dorsal tunnel under the extensor retinaculum, making it an uncommon form of the rare accessorius. PMID- 15835406 TI - Histological findings in titanium implants coated with calcium phosphate ceramics installed in rabbit's tibias. AB - Oral reconstruction using osteointegrated implants are widely indicated nowadays. The implant bone anchorage is very important for its functional stability. Thus, ceramic biomaterials are widely used as coatings of the implant surfaces to accelerate local osteogenesis. The purpose of this study is to assess the biocompatibility and the osteoconduction of two types of calcium phosphate ceramics used as titanium dental implant coatings. These implants were installed in rabbit tibia during an 8-week healing period. The light and fluorescent microscopy observations showed that the materials are biocompatible and that they have osteoconductive activities. PMID- 15835407 TI - Prevention of renal failure. PMID- 15835408 TI - The challenge of kidney transplantation among minority ethnic groups in the UK. AB - The increased rate of renal failure secondary to diabetes among minority ethnic groups compared to Caucasians in the UK has been well documented. However, the impact of this phenomenon on kidney transplant services has been relatively unexplored. The Government has recently published its 'Tackling Health Inequalities: A Programme for Action' report which emphasises the importance of reducing health inequalities at all levels of the health service. This article provides a timely review of the UK's national transplant database examining the provision of kidney transplant services to minority ethnic groups in the United Kingdom. It seeks to explore the demographics of the database by focusing upon waiting list data, donor data, and recipient data. Inequalities do currently exist in transplant services and the solutions to rectifying this situation are complex. However, the financial and human burden of not addressing these inequalities encourages some immediate action. PMID- 15835409 TI - Public awareness and acceptance of tissue and organ donation. AB - Organ donation is a worldwide problem especially in developing countries due to lack of knowledge about brain death and organ transplantation as well as traditional values and beliefs. Limited data are available concerning public knowledge and awareness of organ donation and transplantation in Turkey. A prospective study was carried out and data (n=774) were gathered from a structured questionnaire. The results indicated that 96.6% of participants had heard of organ donation and transplantation, 91.8% approved donation, and 59.2% would consider donating organs of their own. 62.6% were unaware of the donation law. As 81.7% of participants were interested in having more information on the issue, information booklets covering brain death, donation law and the organ donation process may increase donation of organs. PMID- 15835410 TI - Living kidney donor experiences: implications for counselling. AB - This study adds to previous, mostly quantitative, investigation into the experiences of living related kidney donors. Such investigation is important so that potential donors are supported effectively and donation programmes remain relevant and specific to need. Exploration takes place into donor decision-making processes and the most effective forms of professional support. A non-probability sampling technique highlighted eight living related kidney donors who were interviewed using a semi-structured interview format. Raw data was analysed through the qualitative technique of Interpretative Phenomenological Analysis. The decision to donate is made rapidly, decisively and rationally. Professional support provides reassurance to donors, particularly when experiencing acute psychological reaction. The need to provide support to the parents of living donors is specifically highlighted. A comprehensive range of Master Themes are generated through Interpretative Phenomenological Analysis and these reflect the complexity of the donation experience. This suggests donors are concerned with the management of psychological experience rather than with reviewing the appropriateness of an original decision to donate. Evidence indicates that concepts of attitude and self-efficacy belief can develop understanding of the psychological experience of being a living kidney donor. A counselling perspective, with Social Cognitive Theory at its core, is highlighted as a valid method for providing professional support to donors before and after surgery. PMID- 15835411 TI - An integrated vascular management programme. AB - This paper discusses the development of an integrated approach for the creation and maintenance of arteriovenous access (AVA), under the direction of a dedicated vascular access surgeon in close cooperation with the haemodialysis staff, involving pre-operative imaging, anaesthetic and surgical techniques together with a post-operative graft surveillance programme, in order to maximize autogenous arteriovenous access (AAVA) construction (the preferred access for haemodialysis patients with end-stage renal disease), and to improve patency rates for AAVA and prosthetic arteriovenous access (PAVA). PMID- 15835412 TI - Pain degree and skin damage during arterio-venous fistula puncture. AB - The aims of this study were to evaluate the effect of needle bevel position on the degree of pain and damage to the skin covering the vein, in an arteriovenous fistula puncture, in haemodialysis patients. 48 patients with autologous arteriovenous fistula were studied. After puncture the patient was asked about the degree of pain perceived by means of an analogue visual scale and a descriptive verbal scale. When the needle was removed, the length of the cut made by the puncture was measured. The perceived pain assessed by analogue visual scale was greatest when the needle was punctured with the bevel facing upwards rather than downwards (median: 3 versus 2, p<0.003). The prick in the skin was greater when the puncture was made with the bevel facing upwards (19.7 +/- 5.6) rather than downwards (16.2 +/- 3.8, p<0.0001). It is concluded that arteriovenous fistula puncture with the bevel facing downward significantly reduces the degree of pain and the skin lesion at the point of puncture, without increasing the number of punctures. PMID- 15835413 TI - A descriptive analysis of sleep disturbances in patients with end-stage renal disease undergoing haemodialysis. AB - Many studies reveal the presence of sleep disturbances in patients with end-stage renal disease (ESRD) undergoing haemodialysis. The objective of this study was to identify the incidence of sleep disturbances in a sample of 112 patients, being treated by a dialysis centre. Patients who fulfilled the inclusion criteria, were interviewed by means of a questionnaire. The average age was 63.97, 79 male (70.5%) and 33 female (29.4%). The principal pathology underlying subjects' ESRD was hypertension (27.6%) and cardiovascular pathologies were the accompanying pathologies most suffered (75.8%). Sleep disturbances were noted in 54 patients (48.2%). These findings confirm the results of the literature, but less daytime sleepiness and nighttime waking occurred with this study group. PMID- 15835414 TI - Nursing care for dialysis patients in Japan. AB - In Japan, the number of chronic renal failure patients requiring dialysis treatment is increasing by approximately 10,000 patients a year, totalling 229,538 or 1,801.2 patients per one million population at the end of December, 2002 according to the survey conducted by the Japanese Society for Dialysis Therapy. The primary disease is diabetic nephropathy (28.1% of patients), and with greater numbers of long-term and elderly patients, the incidence and severity of complications continue to increase. Dialysis units provide more diversified care, but budget cuts in health care make it difficult to secure sufficient personnel, and the workload on nurses has intensified. In this study, a survey of 157 out of 300 randomly selected dialysis facilities in Japan was made regarding nursing systems and nurses' concerns for medical care of dialysis patients. PMID- 15835415 TI - Late somatic and psycho-social consequences of renal insufficiency in children. AB - The aim of the project was to establish the somatic and psycho-social consequences in adulthood of paediatric end-stage renal disease (ESRD). A long term follow up study of all Dutch patients who started renal replacement therapy at age 0-15 years between 1972 and 1992 and who were born before 1979 was undertaken. The study was performed between 1998 and 2000. About 25% of patients had died at time of investigation. Cardiovascular abnormalities were prominent in both transplanted and dialysis patients and cardiovascular disease was also found to be the most prominent cause of death. Skeletal disorders as a result of metabolic bone disease accounted for most frequent occurring disabilities. The risk for malignancies was 10-fold that of the normal population. Mean IQ and educational attainment were low as compared to the normal population. The overall quality of life was good, despite the co-morbidity and disabilities. Adverse outcomes were in nearly all cases related to a prolonged period of dialysis in the past and to (current) hypertension, not to the state of dialysis at time of investigation. PMID- 15835416 TI - An investigation into stress levels amongst renal nurses. AB - A qualitative study was designed to explore renal nurses' perception of stress within their unit. This study collected information from ten staff nurses through semi structured taped interviews. Using content analysis, interviews were coded and categories and themes were identified. Findings indicate that stress was derived from the following categories: job content, resource issues, professional concerns, professional working relationships, extrinsic factors, and coping mechanisms. The actual words of the nurses were included to provide depth and substance to the categories and themes. A discussion was held regarding the relationship of these findings to theory. These findings have implications for nursing practice, administration and education. Recommendations are also made for future studies. PMID- 15835417 TI - Our dilemma: the self-marginal people. AB - Cultural diversity is nowadays a common phenomenon in most developed countries. Its impact is felt in daily life, as well as in health care settings. Hospital del Mar, due to its geographical location, receives some of the 34.7% of immigrants from a varied cultural background. Endeavours are made to promote cultural awareness among staff and to facilitate communication. However, a Spanish Gypsy group, albeit not immigrants, are by large the problematic cultural group. A questionnaire study was conducted to validate the difficulty in treating these Gypsies versus migrant patients. The questionnaire was distributed to different health care professionals in the hospital. The result is not surprising. The problems encountered with the migrant patients are mainly due to language barrier. Concerning these Gypsies who speak fluent Spanish, the problems are by far more extensive. These are often generated by their abiding cultural values, distinctive lifestyle, low educational level and rejection of the shared norms of the majority. Any attempt to encourage adherence to hospital rules are often futile as a result of the gypsies' negative attitudes. Staff have expressed occasional feelings of weariness when dealing with such situations. PMID- 15835418 TI - Establishing nephrology nursing specialization in Japan: a matter of urgency. AB - This paper explores the current situation regarding the provision of renal services to patients with end-stage renal disease in Japan. The statistical data reveal that Japan is in a predicament regarding the current rate of incidence of end-stage renal disease in the population. The discussion highlights the fact that nephrology nursing practice is as yet not recognised as a speciality in its own right in Japan. Therefore Japanese nurses who work in the field of nephrology are still struggling to establish their own professional identity and to improve the professional level of their practice. This situation persists in Japan despite support from the literature, which confirms that appropriately qualified nephrology nurses are well equipped to provide specialised care. The recent decision by the Japanese Association of Nephrology Nurses is to establish the position of the specialised dialysis nurse, which does not cover the area of kidney transplantation and will start in 2004. It is expected that it will be an important challenge to improve professional skills and nephrology nursing status in Japan. PMID- 15835419 TI - The difference between two means: is it significant? PMID- 15835420 TI - Perspective is everything. PMID- 15835421 TI - Bond strength of composite to dentin using self-etching adhesive systems. AB - PURPOSE: To determine the shear bond strength to dentin of five newer-generation self-etching adhesive systems. METHOD AND MATERIALS: Flat bonding sites were prepared on 100 extracted human molar teeth with 600-grit silicon carbide paper, which exposed the dentin. The systems tested were: Clearfil SE Bond; Optibond Solo Plus Self-Etch; Xeno CF III; Tyrian SPE; and Adper Prompt L-Pop. Following application of each adhesive system, bonded assemblies of Z-100 were prepared using a gelatin capsule matrix (n = 20). Ten specimens in each group were debonded after water storage for 24 hours at 37 degrees C, and 10 were stored for at least 30 days, followed by thermocycling for 850 cycles between water baths at 5 degrees C and 55 degrees C, with a dwell time of 1 minute in each bath. Specimens were loaded to failure in an Instron Model 1123 testing machine with a crosshead speed of 5 mm/min. Statistical analysis included a two-way analysis of variance (adhesive and time) and Tukey's post hoc test. RESULTS: Mean shear bond strengths (MPa) at 24 hours were: Clearfil, 26.4+/-4.2; Optibond, 23.5+/-4.4; Xeno, 19.5+/-4.2; Tyrian, 16.5+/-3.7; and Adper, 15.3+/-3.3. Mean shear bond strengths (MPa) after storage and thermocycling were: Optibond, 27.5+/-5.7; Clearfil, 25.5+/-4.9; Adper, 21.5+/-3.3; Xeno, 20.4+/-2.2; and Tyrian, 19.2+/ 6.4. Both adhesive system and storage time were significant factors affecting shear bond strength (P < .05). CONCLUSION: There are significant differences in the in vitro dentin shear bond strength among the self-etching adhesive materials tested. Water storage and thermocycling did not significantly degrade the shear bond strength of the materials in this study. PMID- 15835422 TI - Hypodontia--a retrospective review of prevalence and etiology. Part I. AB - A retrospective literature review was carried out to determine the prevalence of hypodontia and current knowledge of associated etiological factors. Reported studies vary widely in their reports of prevalence. The majority are retrospective radiographic studies, and reported rates vary from 2.6% to 11.3%. Racial differences have been determined: in Caucasian studies the mandibular second premolars and maxillary lateral incisors were the most frequently recorded absent and in Asian studies, the mandibular incisor was the most frequently recorded absent. Prevalence rates are higher in females compared to males (3:2, respectively). The association of hypodontia with other systemic conditions and dental anomalies is widely reported. Hypodontia can be regarded as a multifactorial condition. Recent developments in molecular genetics have established the importance of the muscle specific homeobox genes (Msx1 and Msx2) in dental development. Specific genes that are responsible for the more severe types of hypodontia, including ectodermal dysplasia, have been identified by linkage analysis. However, variable expression of the trait suggests a polygenic mode of inheritence with epistatic genes and environmental factors interacting. PMID- 15835423 TI - The comparison of a dentin adhesive with calcium hydroxide as a pulp-capping agent on the exposed pulps of human and sheep teeth. AB - OBJECTIVE: This study evaluated the biocompatibility of a one-step dentin bonding agent (Prime&Bond 2.1) in pulp capping compared with calcium hydroxide [Ca(OH)2]. METHOD AND MATERIALS: Thirty sheep teeth and 20 intact human premolars were used. After cavity preparation, pulp exposure was achieved with a bur (#390). Adhesive pulp capping was performed in 25 teeth (15 sheep and 10 human). In the control group (12 sheep and 10 human teeth), pulps were capped with Ca(OH)2 and all of the cavities in both groups were sealed with resin composite. Three of the sheep teeth were used as intact controls. Teeth were extracted 7 or 90 days following treatment and prepared for histological examination and bacterial detection. RESULTS: At 7 days, severe inflammatory responses underlying the bonding agent and in the coronal pulp were observed with soft tissue disorganization in both human and sheep teeth capped with Prime&Bond 2.1. All of the teeth capped with Ca(OH)2 exhibited mild inflammatory reactions limited with the perforation area. After 90 days with the bonding agent, in 3 of 9 sheep teeth, chronic inflammatory reactions were significant, while slight pulpal reactions were observed in the others and dentin bridge formation in all of the sheep teeth was found. However, in human pulps, persistent, unresolved inflammation with the lack of dentin bridge formation was observed. In the Ca(OH)2 group, pulp repair with dentin bridging was found in all of the teeth, both sheep and human. No correlation was found between the presence of inflammation and bacterial staining using Spearman rank correlation test (P > .05). CONCLUSION: Prime&Bond 2.1 facilitates enhanced pulp healing and bridge formation in sheep teeth, but in human teeth it was not as successful as Ca(OH)2 as a pulp capping agent. PMID- 15835424 TI - Labial-cervical-vertical groove in maxillary permanent incisors--prevalence, severity, and affected soft tissue. AB - OBJECTIVES: To investigate the prevalence and severity of a labial-cervical vertical groove (LCVG) in maxillary permanent incisors and its effect on the associated gingival tissue. METHODS: A total of 600 adolescents (293 boys and 307 girls, mean age 13.6+/-1.99 years) were randomly selected and examined for the presence of LCVG. The deformity was classified as mild, moderate, or severe according to predetermined criteria. Gingival coverage at the groove site was defined as normal, partial, and irregular. RESULTS: LCVG was found in 27 adolescents (4.5%). It was unilateral in 24 (89%) and bilateral in 3 (11%). The ratio of central to lateral incisors was 29:1. No sexual dimorphism or side prevalence were found. Mild LCVG was found in 22 incisors, moderate LCVG in 7 incisors, and severe LCVG in one incisor. Moderate LCVG was 5 to 6 times more susceptible to partial or irregular coverage of the gingival margin than mild LCVG. The gingival sulcus in teeth with LCVG demonstrated a significant (P = .001) increase in depth compared to non-LCVG teeth (1.55+/-0.90 mm vs 1.18+/-0.75 mm). CONCLUSIONS: An LCVG is a deformity confined predominantly to a single permanent maxillary central incisor. Its prevalence is not connected with gender. Most LCVGs are mild and often difficult to detect. However, the greater the severity, the more gingival irregularity is associated. This and the increase in sulcus depth in LCVG incisors are adverse predispositions for periodontal sequelae, calling for cautious oral hygiene maintenance. PMID- 15835425 TI - Nutrition & oral health. Dietary supplements and Oral Health: should the dentist ask? PMID- 15835426 TI - Oral findings in patients with obstructive sleep apnea syndrome. AB - OBJECTIVE: To evaluate the role of dental parameters that may contribute in increasing apneic activity in patients with obstructive sleep apnea syndrome (OSAS). METHOD AND MATERIALS: A total of 152 subjects (121 males and 31 females ) were examined from October 1999 until September 2001, for sleep disorder symptoms. All patients underwent polysomnography. Any abnormal or unusual features that could inhibit the upper airway of the oral cavity were evaluated by an oral pathologist. RESULTS: Statistical analysis of the independent structural variables and the respiratory disturbance index proved to be significant only in the cases of retropositioned or narrow hard palate with a vertically positioned soft palate, the type of breathing (oral breathing), and the enlarged uvula. CONCLUSION: The oral soft tissues seem to be more closely associated with OSAS. Therefore, the diagnosis of the structural features of the oral cavity by the clinician is useful to predict apneic activity. PMID- 15835427 TI - Tissue destruction in periodontitis: bacteria or cytokines fault? AB - The pathogenesis of periodontal disease involves the sequential activation of a great variety of components of the host immune response, primarily acting to defend periodontal tissues against bacterial aggression, but also functioning as mediators of tissue destruction. The expression of the disease results from the interaction of host, microbiological agents, and environmental factors. Leukocytes play a critical role in the pathogenesis of the disease, producing different cytokines, chemokines, and other mediators, thus generating a host defense response, as well as inducing tissue inflammation and bone destruction. The aim of this review is to address the role of some inflammatory mediators in response to bacterial aggression in periodontitis. PMID- 15835428 TI - Oral manifestations during chemotherapy for acute lymphoblastic leukemia: a case report. AB - A 14-year-old, male patient was referred for the treatment of mucositis, idiopathic facial asymmetry, and candidiasis. The patient had been undergoing chemotherapy for 5 years for acute lymphoblastic leukemia. He presented with a swollen face, fever, and generalized symptomatology in the mouth with burning. On physical examination, general signs of poor health, paleness, malnutrition, and jaundice were observed. The extraoral clinical examination showed edema on the right side of the face and cutaneous erythema. On intraoral clinical examination, generalized ulcers with extensive necrosis on the hard palate mucosa were observed, extending to the posterior region. Both free and attached gingivae were ulcerated and edematous with exudation and spontaneous bleeding, mainly in the superior and inferior anterior teeth region. The tongue had no papillae and was coated, due to poor oral hygiene. The patient also presented with carious white lesions and enamel hypoplasia, mouth opening limitation, and foul odor. After exfoliative cytology of the affected areas, the diagnosis was mixed infection by Candida albicans and bacteria. Recommended treatment was antibiotics and antifungal administration, periodontal prophylaxis, topical application of fluor 1.23%, and orientation on and control of proper oral hygiene and diet during the remission phase of the disease. PMID- 15835429 TI - Clinical images in oral medicine. Erythema multiforme. PMID- 15835430 TI - Radiographic diagnostics. Radicular cyst. PMID- 15835431 TI - Prepare to perform. PMID- 15835432 TI - 'Nursing is the most emotionally rewarding career'. AB - This artie presents a brand manifesto for the profession, based on work carried out by consultants Red Spider as part of our Nursing the Future campaign. We have also used the 'redprint' to commission a film that promotes nursing and midwifery. How the work was carried out: --Interviews with a number of experienced 'stakeholder' nurses. --A one-day workshop involving experts from a variety of fields. Findings include: --Nursing needs a modern image. --Nursing is the most emotionally rewarding career. PMID- 15835433 TI - Keeping watch. AB - Early detection of heart attack means that potentially life saving treatment can begin immediately. HealthWatch offers a distant monitoring service via a call centre run by nurses and doctors to offer reassurance or enable early A&E attendance if required. PMID- 15835434 TI - Humility heals. PMID- 15835435 TI - Dear CNO. PMID- 15835436 TI - Hold the line. PMID- 15835437 TI - Developing a new generation of nurse entrepreneurs. AB - This article reviews the policy implications associated with the creation of nurse entrepreneurs, the levels of progress achieved and the major obstacles to developing these roles. International experience and the cultural climate of the NHS in the U.K. are also discussed. The way in which entrepreneurial roles are evolving within the NHS and the future implications and developments for the wider not-for-profit and private sectors are explored. PMID- 15835438 TI - Supplementary prescribing in mental health and learning disabilities. AB - Nurse prescribing in mental health and learning disability services is a new development. The experiences of nine nurses working in mental health and learning disabilities, who formed part of the first cohort in the U.K. to undertake the supplementary nurse prescribing course, are described. Experiences of the course and implementation of supplementary prescribing in practice are discussed. The attitudes of nurses, other health professionals and patients to nurse prescribing are also explored. PMID- 15835439 TI - An introduction to behaviour change among clients. AB - This article, the first in the series of eight articles on health promotion, introduces nurses in clinical practice to the principles and practice of facilitating behaviour change among clients. The series will contain information on strategies to promote and support change in adolescents, pregnant women and vulnerable groups. PMID- 15835440 TI - Effects of nightlife activilty on health. AB - This article aims to increase understanding of nightlife health and the potential for nurses to reduce some of the negative health consequences associated with nightlife activity. PMID- 15835441 TI - Thinking about therapies. PMID- 15835442 TI - CAM in women's health: safely navigate the waters. PMID- 15835443 TI - It's not about food: getting to the heart of eating disorders. PMID- 15835444 TI - Embracing the aging process. PMID- 15835445 TI - Perimenopause: helping women be heard. PMID- 15835446 TI - Women behind bars: the challenge to care. PMID- 15835447 TI - From dream to reality: how a parish nurse program is born. PMID- 15835448 TI - When God calls you to a hard place. Why start when there is such a need? PMID- 15835449 TI - The cost of a little kindness. PMID- 15835450 TI - How does spiritual care begin? PMID- 15835452 TI - Meperidine (Demerol) safety issues. PMID- 15835453 TI - Neostigmine: an alternative treatment for constipation. AB - Constipation is a common complication of amyotrophic lateral sclerosis (ALS), especially as the disease progresses. While ALS patients may experience disturbed gastrointestinal motility due to the nature of the disease and decreased physical activity, the constipation is not usually caused by mechanical obstruction. Acute colonic pseudo-obstruction (ACPO) is a syndrome characterized by massive dilation of the colon without mechanical obstruction. Recent studies have shown neostigmine may be an effective treatment for ACPO. Through a case study, the author discusses the use of neostigmine and its nursing implications on a patient with ALS. PMID- 15835454 TI - Can minimal patient restraint be safely implemented in the intensive care unit? AB - The introduction of a corporate policy on the minimal use of restraints created concern within our intensive care unit. It was thought that corporate leaders did not understand the necessity of patient restraints within critical care. In an effort to make a stand against the developing policy, a group of critical care nurses reviewed the literature for evidence to support the use of restraints?with this patient population. The evidence that was found surprised the group and created the impetus to construct tools and a learning package to support minimal restraint within the intensive care unit. The results after one year demonstrate that minimal restraint practices can be safely implemented in intensive care using a comprehensive program; a program that effects a paradigm shift from caregiver paternalism to one that supports the patient's dignity of risk. PMID- 15835456 TI - [Happy-ending stories]. PMID- 15835455 TI - Knowledge and skill acquisition for critical care nursing practice. AB - In many critical care units, extended orientation or education programs have been offered to prepare the registered nurse to work in this specialty area. Current shortages of nurses and less experienced nurses applying for critical care positions have augmented the need for educational programs. In the Edmonton region, a partnership between two public institutions led to the development and implementation of an innovative educational program. In this article, we describe challenges and strategies to prepare critical care nurses to integrate knowledge, and meet the standards and expectations of both partners. In particular, methods are described to achieve theoretical and clinical outcomes. This article contributes to the discussion and implementation to offer knowledge and skill acquisition for a novice critical care nurse. PMID- 15835457 TI - [Slices of hospital life. New culture and quality of cooperation]. PMID- 15835458 TI - [Aftercare concept for psychiatric nurses. From here we achieve the first success]. PMID- 15835459 TI - [The failure or terminal care nurses. The judgment is admitted--the problem remains]. PMID- 15835460 TI - [Hospital information systems. When th information disrupts the regulations]. PMID- 15835461 TI - [Care and control in a social therapy home. The "astonishing question" unlocks the obvious]. PMID- 15835462 TI - [Self-care in diabetes care. Knowledge is not enough]. PMID- 15835463 TI - [The personnel situation in large hospitals]. PMID- 15835464 TI - [Premature birth. The skin needs special care]. PMID- 15835465 TI - [Thanks to Mr. Theophraste]. PMID- 15835466 TI - [Care of schizophrenic patients. Turn over the disease to the patient]. PMID- 15835467 TI - [Health promotion in the Geneva area. Concerning bad manners and friendliness]. PMID- 15835468 TI - [A way to age and to care for oneself]. PMID- 15835469 TI - [The paternal role in the family. Honor the father!]. PMID- 15835470 TI - [Development of palliative care for the aged. An approach founded in authenticity]. PMID- 15835471 TI - [Experiencing the relations with a person at the end of life. "In the relationship, all care is guided by the heart]. PMID- 15835472 TI - [Home care services and. An important consideration of territoriality]. PMID- 15835473 TI - [Prematurity. The sick premature infant need particular protection]. PMID- 15835474 TI - What goes around comes around? PMID- 15835475 TI - Kernicterus: still a concern. AB - The presence of yellow staining and damage to the brain caused by unconjugated bilirubin was first described by Hervieux in 1847. Kernicterus, the technical term used to describe the intense yellow staining in the basal ganglia of the brain, was first used by Schmorl in 1903. Perhaps as many as 60 percent of all babies born each year in the U.S. are diagnosed with clinical jaundice. Kernicterus is a preventable brain injury caused by severe jaundice, yet it remains a threat today. Because of this, anyone caring for newborns must be aware of the risks and treatment for hyperbilirubinemia and the sequelae of this seemingly benign entity. PMID- 15835476 TI - Idiopathic dilation of the ductus venosus: an unusual anomaly of the fetal blood vessels. AB - The ductus venosus (DV) is a blood vessel unique to the fetal circulation. It functions as a shunt between the umbilical vein and inferior vena cava by passing the hepatic microcirculation and shunting highly oxygenated blood to the brain and myocardium. The DV has received less attention than the ductus arteriosus; however, with modern ultrasonographic imaging, the DV is easily identified and is becoming a valuable diagnostic tool in the areas of maternal-fetal medicine. We report a rare case of idiopathic dilation of the DV that was diagnosed during routine ultrasounds. The presence of blood flow turbulence within the dilated area and the concern that blood vessels might rupture resulted in elective cesarean section at 34 weeks gestation. The infant was discharged after 14 days. Dilation of the DV, its size as well as the presence of turbulence in the dilated area might be important factors in determining the time of delivery and could contribute to a better outcome. PMID- 15835477 TI - Car seat safety: literature review. AB - After staggering numbers of infants were killed in automotive crashes in the 1970s, the American Academy of Pediatrics (AAP) recommended in 1974 universal use of car seats for all infants. However, positional problems were reported when car seats are used with premature infants less than 37 weeks gestational age as a result of head slouching and its sequelae. In 1990, the AAP responded with another policy statement introducing car seat testing. It recommended that any infant at or under 37 weeks gestational age be observed in a car seat prior to discharge from the hospital. The AAP did not give specific guidelines on type of car seat, length of testing, equipment, or personnel proficiency, however. Few nurseries have standard policies to evaluate car seats, to teach parents about car seats, or to position newborns in them, and not all hospitals actually conduct car seat challenges or have common standards for testing that is performed. PMID- 15835478 TI - A comparison of the light-reduction capacity of commonly used incubator covers. AB - PURPOSE: The use of incubator covers to enhance preterm infants' rest and recovery is common in the NICU. However, the kinds of covers used vary extensively among and within nurseries. Few data exist on the effectiveness of different types of covers in reducing light levels to the infant. This study compared several types of commonly used incubator covers as to efficacy of light reduction. DESIGN: A descriptive, comparative design was used in this study. SAMPLE: Twenty-three incubator covers were tested, including professional, receiving blanket, hand-crocheted, three-layer quilt, and flannel. MAIN OUTCOME VARIABLE: The percentage of light level reduction of different incubator covers under various ambient light level settings. RESULTS: The amount of light reduction provided by incubator covers varies depending on type of fabric as well as percentage of incubator surface shielded by the cover. Dark-colored covers provided greater light reduction than bright/light-colored covers when covers identical in fabric type were compared. The light-reduction efficiency of the covers varied depending on the level of ambient light. Covers provided less light reduction in higher ambient light levels. PMID- 15835479 TI - Gastroesophageal reflux and prokinetic agents. PMID- 15835481 TI - Progress in the elimination of leprosy in India. PMID- 15835480 TI - Getting high on performance! AB - The success of the NICU depends in large part on the manager's ability to continuously identify the high, middle, and low performers on the unit. To retain top performers, the manager must ensure a challenging work environment paired with zero tolerance for substandard performance. Middle performers need positive recognition and the opportunity to stretch for improved performance. Once employees have been categorized, the manager needs to consistently develop performance improvement plans for each one. PMID- 15835482 TI - Trends in tuberculosis research. PMID- 15835483 TI - Protective role of BCG vaccination against tuberculous meningitis in Indian children: a reappraisal. AB - BACKGROUND: The protective role of BCG vaccination against tuberculous meningitis (TBM) is still controversial in India, largely due to the presence of other predisposing factors such as age, nutritional and socioeconomic status, and household contact. Very few Indian studies have focused on the interaction between BCG and these factors on the occurrence of TBM. METHODS: We did an unmatched hospital-based case-control study with prospective enrolment. Children with TBM diagnosed on the basis of predefined criteria were enrolled as cases. For each case, two children admitted on the same day and who did not have any neurological symptoms were enrolled as controls. Demographic data and information on predisposing factors for tuberculosis were collected for both cases and controls and the presence of a BCG scar was charted. Cases and controls were compared by univariate followed by multivariate analysis to obtain significant independent predictors for the occurrence of TBM. To assess the interaction between other predisposing factors and protective efficacy of BCG, a stratified analysis was also done. RESULTS: A total of 91 cases and 182 controls were enrolled over a one-year study period, of which 37 cases and 111 controls had a BCG scar. The crude odds ratio for the occurrence of TBM in the absence of a BCG scar was 2.28 (range: 1.32-3.94). The time elapsed since vaccination was significantly longer in the cases. Also, the proportion with a household contact was significantly higher in the cases, the mean age of the cases was higher than that of the controls, and the mean weight and height for age percentage were significantly lower. The cases had a significantly lower socioeconomic status. On multivariate analysis, the significant independent predictors for the occurrence of TBM were positive household contact with tuberculosis (adjusted OR 4.26; 95% CCI 2.26-8.04), absent BCG scar (adjusted OR 1.98; 95%ClI 1.09-3.57) and rural residence (adjusted OR 2.07; 95% ClI 1.02-4.17). CONCLUSION: Vaccination with BCG was found to be protective even after controlling for the effect of other variables. Stratified analysis showed that protection due to BCG failed to reach significance for those > 5 years of age, if the weight was <6 0% of that expected for age, in the presence of a household contact with tuberculosis, and in socioeconomic classes III, IV and V. PMID- 15835484 TI - Outcome of acute myeloid leukaemia in adults: a retrospective analysis. AB - BACKGROUND: There are little data from India on the management of acute myeloid leukaemia. With better understanding of the biology of the disease, and routine use of high-dose cytarabine as post-remission therapy with or without haematopoietic blood stem cell transplantation (HSCT), the results have improved in the past two decades. We analysed our results in a cohort of recently treated patients. METHODS: A total of 166 newly diagnosed patients with AML (excluding acute promyelocytic leukaemia), 15-60 years of age were treated with daunorubicin (60 mg/m2/day x3 days) or idarubicin (12 mg/m2/day x3 days) with cytarabine (100 mg/m2/day continuous i.v. infusion x7 days) induction chemotherapy. Post remission therapy included 2 cycles of high-dose cytarabine (15-18 g/m2) followed by monthly cycles of outpatient maintenance chemotherapy x4 cycles, consisting of daunorubicin (45 mg/m2 i.v. x1 day and cytarabine 100 mg/ m2 s.c. twice daily x5 days). Six patients in remission received sibling donor allogeneic HSCT. RESULTS: Morphological complete remission was achieved in 69.9% of the patients. Resistant disease after induction chemotherapy was seen in 14.6% and early mortality occurred in 16%. Relapse-free survival and event-free survival at a median of 36 months was 34% and 22%, respectively. Relapse occurred in 43.9%. The median duration of remission was 12 months. CONCLUSIONS: Our results conform to the published literature from larger cooperative studies from the West. Currently available cytotoxic drugs are unlikely to improve the results any further. PMID- 15835485 TI - Community prevalence of antibodies to human immunodeficiency virus in rural and urban Vellore, Tamil Nadu. AB - BACKGROUND: Human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) are becoming increasingly common in India. Currently, antenatal prevalence is a surrogate marker for HIV prevalence in the community. The association between antenatal and community prevalence of HIV needs to be validated so that estimates can be verified or adjusted appropriately. METHODS: A probability proportional to size cluster survey was conducted in the Kaniyambadi block of Vellore district and in the urban wards of Vellore town to estimate the prevalence of antibodies to rubella from August 1999 to February 2000. All personal identifier data from the serum samples were removed to yield a collection for which only the age and sex were known. Estimation of antibodies to HIV in sera from individuals between 15 and 40 years of age, was carried out by one screening ELISA and the reactive sera were further subjected to a supplementary test. RESULTS: We tested 1512 serum samples from subjects residing in rural areas and 1358 samples from those residing in urban areas. The seropositivity among rural samples was 0.66% and among urban samples 1.4%. The prevalence was almost equal among men and women and the youngest infected individual was 15 years old. CONCLUSION: The prevalence of HIV during the period of study was similar to the national surveillance data for Tamil Nadu based on antenatal women. HIV prevalence differs in urban and rural Tamil Nadu, with urban areas having a higher burden of the disease. PMID- 15835486 TI - The first successful simultaneous pancreas-kidney transplant in India. AB - Insulin-dependent diabetes mellitus is associated with renal failure, diabetic retinopathy, neuropathy and vasculopathy. We report the first successful simultaneous pancreas-kidney transplant in India in a young diabetic with renal failure. The dual transplant has cured his diabetes and renal failure and has had a beneficial effect on his neuropathy, retinopathy and quality of life. Obstacles to dual transplant in India include a lack of suitable recipients and a cadaver donor programme that is still in its infancy. PMID- 15835487 TI - Kawasaki disease. AB - Kawasaki disease (KD) is a common vasculitic disorder usually seen in children below 5 years of age. The disease can present with protean clinical manifestations which include high grade fever (for at least 5 days), rash, redness of the lips and a typical strawberry tongue, cervical lymph node enlargement (often unilateral), swelling over the hands/feet and, later a characteristic peripheral desquamation over the fingers and toes. These clinical features appear sequentially and the findings may change from day-to-day. Thus, all these features may not be seen together at any one point of time. The diagnosis rests on the recognition of this characteristic temporal sequence of clinical events, none of which are, by themselves, pathognomonic. Establishing a diagnosis of KD may be further complicated by the occurrence of several other, seemingly unrelated, clinical features. These include irritability, neck stiffness, sterile pyuria, pneumonitis, hydrops of the gallbladder and hepatitis among many others. There is no laboratory test that can help in confirming a diagnosis of KD. Left untreated, up to 20% of children with KD can develop coronary aneurysms with catastrophic long term sequelae. It is important to diagnose KD in the first 10 days of the illness so that appropriate therapy with intravenous immunoglobulin and aspirin can be Initiated. All paediatricians, and physicians looking after children, need to be aware of this condition which is now being increasingly recognized in India. PMID- 15835488 TI - Corticosteroid use in head injuries. PMID- 15835489 TI - Cost and efficiency of HIV voluntary counselling and testing centres in Andhra Pradesh, India. AB - BACKGROUND: [corrected] As part of the effort to control HIV/AIDS, the number of HlV voluntarycounselling and testingcentres (VCTCs) is increasing rapidly in the public health system of the Indian state of Andhra Pradesh, which is estimated to have one of the highest rates of HIV infection in India. However, systematic data on the cost and efficiency of providing VCT services in India are not available to help guide efficient use of resources for these services. METHODS: We used standardized methods to obtain detailed cost and output data for the 2002-03 fiscal year from written records and interviews in 17 VCTCs in the public health system in Andhra Pradesh. We calculated the economic cost per client receiving VCT services, and analysed the variation and determinants of total and unit costs across VCTCs. We used multivariate regression techniques to estimate incremental unit costs. We assessed hurdles towards serving an optimal number of clients by VCTCs. RESULTS: In the 2002-03 fiscal year, 32 413 clients received the complete sequence of services at the 17 VCTCs, including post-HIV test counselling. The number of clients served by each VCTC ranged from 334 to 7802 (median 979). The overall HIV-positive rate in post-test counselled clients was 20.5% (range 5.4% 52.6%). The cost per client for the complete VCT sequence varied 6-fold between VCTCs (range Rs 141.5-829.6 [US 2.92-17.14 dollars], median Rs 363.5 [US 7.51 dollars]). The cost per client was significantly lower at VCTCs with more clients (p < 0.001, R2 = 0.83; power function) due to substantial fixed costs. Personnel made up the largest component of cost (53.7%). The cost per client had a significant direct relation with percent personnel cost for VCTCs (p < 0.001, R2 = 0.58; exponential function). A multiple regression model revealed that the incremental cost of providing complete VCT services to each HIV-positive and negative client was Rs 123.5 (US 2.54 dollars) and Rs 59.2 (US 1.22 dollars), respectively. Fourteen VCTCs (82.4%) reported that they could serve more clients with the available personnel and infrastructure, and that inadequate demand for their services was the main hurdle towards achieving this. CONCLUSION: These data suggest that the efforts of the National AIDS Control Organisation of India and the Andhra Pradesh State AIDS Control Society in increasing VCTCs could yield even higher benefit if the demand for these services was enhanced, as this would increase the number of clients served and reduce the cost per client. Ongoing systematic cost-efficiency analysis is necessary to help guide efficient use of HIV-control resources in India. PMID- 15835490 TI - Urban private practitioners: potential partners in the care of patients with HIV/AIDS. AB - BACKGROUND: The private medical sector is an important source of healthcare in India. Increasingly, concerns have been raised about its role in the care of patients with HIV/AIDS. Evidence about private practitioners' existing management practices will help to create policies addressing this sector. METHODS: A central urban area of Pune city was selected for its high density of healthcare facilities. Private practitioners in the area were interviewed using a structured interview schedule. Based on a 1-year recall period, the schedule covered different aspects of the practitioners' HIV/AIDS management practices including diagnosis, treatment and referral. RESULTS: Of the 215 practitioners interviewed, 66% had tested and diagnosed HIV infection. Fifty-four per cent had been consulted by HIV-infected clients 'shopping' for alternative diagnoses or treatment. Overall, 75% of the respondents had been consulted by HIV-infected clients for treatment. Of these, 14% had prescribed antiretroviral drugs, sometimes without adequate knowledge of the guidelines for their use. Other supportive and symptomatic treatments were also frequently prescribed. Private practitioners commonly referred HIV-infected clients for management to other private doctors, or to public hospitals. There were variations in respondents' practices by sex and system of medicine. CONCLUSION: Private practitioners are actively involved in diagnosing and managing patients with HIV/AIDS. Some of their management practices are inappropriate and need to be remedied. There are also concerns about gaps in the continuity of care of HIV-infected persons, for which networks between providers need to be strengthened. Public-private partnerships must be created to improve the flow of information to private practitioners, and Include them in the national health framework. PMID- 15835491 TI - Geriatric dentistry: the need for a new specialty in India. PMID- 15835492 TI - Integration of online formative assessments into medical education: experience from University of Zagreb Medical School, Croatia. PMID- 15835493 TI - The pound of cure is well worth avoiding. PMID- 15835494 TI - Priapism following oral sildenafil abuse. PMID- 15835495 TI - Genotoxic agents detected by plant bioassays. AB - Seven higher plant species (Allium cepa, Arabidopsis thaliana, Glycine max, Hordeum vulgaris. Tradescantia paludosa, Vicia faba, and Zea mays) were reviewed for their ability to detect genotoxicity of chemical agents under the U.S. Environmental Protection Agency (U.S. EPA) Gene-Tox program in the late 1970s. Six bioassays-Allium and Vicia root tip chromosome breaks, Tradescantia chromosome break, Tradescantia micronucleus, Tradescantia-stamen-hair mutation, and Arabidopsis-mutation bioassays- were established from four plant systems that are currently in use for detecting the genotoxicity of environmental agents. Under the Gene-Tox program, the Crepis capillaris-chromosome-aberration test was added to the existing six bioassays. The current review is limited to chemical agents that exhibit a positive response to any of these seven plant bioassays. From 158 articles reviewed, 84 chemicals were compiled in three categories: carcinogens, clastogens, and mutagens. As none of these plant bioassays can detect tumor initiation or cancerous growth, the chemicals were categorized as carcinogens based on their characteristics defined by the U.S. EPA's Superfund Priority 1 List and/or by the chemical listings of the Sigma and Aldrich Chemical Companies. Certain mutagens were categorized in the same manner in addition to the agents detected as mutagens by these plant bioassays. PMID- 15835496 TI - The epidemiology of pesticide exposure and cancer: A review. AB - Cancer is a multifactorial disease with contributions from genetic, environmental, and lifestyle factors. Pesticide exposure is recognized as an important environmental risk factor associated with cancer development. The epidemiology of pesticide exposure and cancer in humans has been studied globally in various settings. Insecticides, herbicides, and fungicides are associated with hemopoetic cancers, and cancers of the prostate, pancreas, liver, and other body systems. The involvement of pesticides in breast cancer has not yet been determined. In developing countries, sufficient epidemiologic research and evidence is lacking to link pesticide exposure with cancer development. Agricultural and industrial workers are high-risk groups for developing cancer following pesticide exposure. Children of farm workers can be exposed to pesticides through their parents. Maternal exposure to pesticides can pose a health risk to the fetus and the newborn. The organophosphates are most the commonly used compounds, but the organochlorines are still permitted for limited use in developing countries. Pesticide exposure, independently or in synergism with modifiable risk factors, is associated with several types of cancer. PMID- 15835497 TI - Mercury in the environment: effect on health and reproduction. AB - Mercury is a heavy metal that is found naturally in the environment in various forms. Human activity can release mercury into the air, water, and soil. Mercury is also released into the environment after its conversion to methylmercury by bacteria. Mercury was once used in medicine, but the medicinal aspect changed because of its devastating poisoning effect on humans and animals. Today, mercury is one of the most potent neurotoxins known, having a number of adverse health effects in animals and humans. As the sources of mercury are many, the general population is exposed to mercury in day-to-day life, in occupational settings, and in cases of accidental exposure. In addition, ignorance about the use of mercury in cosmetics and religious materials has opened an additional source of exposure. Therefore, making people aware of mercury's effects on health, its sources of entry into the environment, and its chelating remedies becomes a necessity so that strategies can be adopted to minimize use and exposure. PMID- 15835498 TI - The use of national health registers for studying environmental causes of congenital defects. AB - This review summarizes and discusses the use of central health registers in studies of causes of congenital malformations. The review is illustrated using data from the Swedish Medical Birth Register with adjacent registers and gives examples of different methods to identify environmental exposures. The review stresses the usefulness of this approach but also the inherent problems of which the researcher must be fully aware. PMID- 15835499 TI - Anthropological characteristics of Bulgarian newborns. AB - The aim of the study was to assess the specificity of newborns' physical development in Bulgaria at the beginning of the 21st Century. A detailed anthropological research of 219 healthy and full-term newborns (110 boys and 109 girls) was carried out during April-May 2001. In the present study, data about stature, body weight and six standard skinfolds on the body and extremities are analyzed. The relative share of each skinfold regarding the sum of the six studied skinfolds was used to characterize the topical distribution of subcutaneous fat tissue. The sexual differences in the features investigated and their secular changes were evaluated. The results show that even at birth, sexual dimorphism exists in the basic anthropometrical characteristics. Newborn boys were longer and heavier than newborn girls. Girls had thicker skinfolds than boys and a greater relative share of subcutaneous fat tissue on the back and the lower extremities, whereas boys had a greater relative share on the abdomen and the upper extremities. Compared with newborns from past generations, newborns here from both genders had lower mean values of the basic anthropometrical characteristics. PMID- 15835500 TI - Not so NICE. A proposal from the National Institute for Clinical Excellence to withdraw treatments for Alzheimer's disease from the NHS has left support groups reeling. PMID- 15835501 TI - And the winner is... Award-winning nurses who help make every day an older person's day. PMID- 15835502 TI - The abuse of older people. PMID- 15835503 TI - Single assessment for older people: new dawn or continued confusion? PMID- 15835504 TI - Specialist nurse practitioners in mental health: the Shropshire experience. PMID- 15835505 TI - Infections and their treatment in older people. AB - Bacteria are robust micro-organisms that can survive even in the presence of antibiotics. This has led to problems treating bacterial infections. Bacteria are the most common cause of infection and Streptococcus pneumoniae is one such infection. This bacterium causes community-acquired pneumonia in older people and, though immunisation is available, older people continue to die from pneumococcal pneumonia. Penicillin continues to be used as the main method of treatment against Streptococcus pneumoniae but antibiotic resistance is increasing PMID- 15835506 TI - Promoting good care for people with hearing impairment. PMID- 15835507 TI - Automated external defibrillators (AEDs) for out-of-hospital resuscitation: the role of first responders. PMID- 15835508 TI - Stigma and mental illness. PMID- 15835510 TI - COPD: illness severity, resource utilisation and cost. AB - COPD is a major source mortality and morbidity International evidence suggests the associated financial burden substantial. This research was undertaken to capture the costs associated with COPD in an Irish secondary care setting. 150 COPD patients attending a respiratory clinic were interviewed. Data on healthcare utilisation were collected. Prescription data and clinical severity data were collected from patient notes. The average direct cost of COPD was estimated at 4,730 euros. Secondary care costs accounted for 44% of total costs. Lost productivity averaged 668 euros. The main direct cost driver was in-patient admissions. A positive relationship was found between average direct costs and both clinical and symptom severity. The costs of COPD increased with increasing illness severity driven mainly by secondary care in-patient costs. If illness progression can be slowed and symptom severity can be reduced it may be possible to achieve significant reductions in expenditure and free resources within the healthcare system. PMID- 15835509 TI - Antibiotic resistant tuberculosis and bovine tuberculosis in an Irish hospital population (1991 to 2001). AB - Of 864 consecutive culture positive cases of tuberculosis (TB) treated in the period 1991-2001, a total of 19 (2.1%) were M. bovis and 32 (3.7%) were drug resistant - Multi-Drug Resistant Tuberculosis (MDR-TB) (8); poly drug resistant (7); mono drug resistant (17). Iatrogenic factors were important in the cause of resistance in 9 cases (4 cases of MDR-TB): in 6 cases patients factors were important. Comparison with previous report for the period 1982-1985 showed relatively more isolates of M. bovis (p = 0.02); drug resistant TB (p = 0.005); and MDR-TB (p = 0.01) in the current study. Comparison with the background population of TB patients showed that patients with resistant TB were more likely to have a previous history of TB (Relative Risk = 3.58) and more likely to be foreign nationals (Relative Risk = 1.93) We conclude that while overall incidence of TB has declined that in relative terms the caseload of drug resistant TB and MDR-TB has increased. The drug cost of treating MDR-TB may be as high as 44,900 euros per patient. PMID- 15835511 TI - Chronic kidney disease in the elderly; a silent epidemic. AB - Chronic kidney disease has been shown to be associated with significant increases in mortality and morbidity even in early stages. Despite this it is rarely diagnosed, actively investigated or managed in the elderly. We set out to establish the prevalence of CKD and identify causative factors in a consecutive series of referrals to a geriatric medical clinic. We calculated glomerular filtration rates (GFR) for 101 patients attending a geriatric medical clinic using the Cockroft and Gault formula, and collected data on medications and relevant past medical history. Mild CKD (GFR <60ml/min) was present in 80% of the group. Only 10% of these patients had serum creatinine >130(mmol/L. hypertension was present in 50% and only 9% were diabetic. Almost one third were on inappropriate drugs or dosages; most commonly non-steroidal anti-inflammatory drugs (99%). Chronic kidney disease is extremely common in older people attending a geriatric medical clinic and carries significant risks. CKD will not be detected by serum creatinine alone in 90% of cases. Routine calculation of GFR should be used to provide a more accurate measure of renal impairment and allow earlier intervention, risk factor reduction and improve prescribing practices. PMID- 15835512 TI - Guidelines for the medical management of Irish children and adolescents with Down syndrome. AB - Down syndrome (DS) is the most common chromosomal cause of developmental disability in Ireland. Children with DS have a high incidence of associated treatable medical disorders where early intervention carries a better outcome. Currently there are no agreed protocols for the screening and management of children and adults with DS in Ireland. A cross-sectional study of 394 children and adolescents was undertaken in the Eastern Regional Health Authority (ERHA) to assess the medical needs of children and adolescents with DS, in order to develop medical management guidelines. This study provides evidence-based data that children and adolescents with DS have a high incidence of treatable medical disorders, which supports the need for the medical management guidelines presented. PMID- 15835513 TI - Changes in trauma service workload since the introduction of the penalty points system. AB - With the introduction of the penalty points system (PPS) has come many media reports indicating a reduction in road traffic accident (RTA) rates. We hypothesised that reduced RTAs would result in a reduction in hospital trauma workloads. To study this we examined the numbers of RTA related femoral shaft fracture presentations to the six Dublin teaching hospitals and Waterford regional hospital within the six months since the introduction of the PPS and the two previous corresponding six month periods. We also analysed all RTA related discharges from Beaumont hospital during these three time periods. Whilst a slight reduction in RTA related femoral shaft fracture numbers was observed a dramatic reduction in total RTA related discharges from Beaumont hospital was identified with 70 patients discharged in the first six months since the introduction of the PPS compared to 124 and 125 discharges in the same two preceding six month periods. This reduction was greatest for head and thoracic injuries which were halved, while total numbers of limb injuries were maintained, resulting in no reduction in orthopaedic RTA related discharges or workload. PMID- 15835514 TI - Ascertaining women's preferred mode of address and preferred choice of title during pregnancy and childbirth. AB - To determine how women in pregnancy would like to be addressed and to ascertain their preferred choice of title during pregnancy. A questionnaire was administered to 925 antenatal women. Midwifery and medical staff (183) were invited to respond to a similar questionnaire. The response rate was 71.2% from the survey of pregnant women. The vast majority (82.1%) preferred to be addressed by their first name. Women were in favour of being called 'patient' (32.8%) as their first choice. The staff survey yielded a response rate of 77%. The majority (81.8%) of health professionals preferred to address women by their first name. 'Mother' (28.7%) was the most popular first choice. We conclude that women in pregnancy do have a preference on how they would like to be addressed and this is predominantly by first name. Health professionals also prefer to call pregnant women by their first name. The term 'patient' was the most popular first choice of title of women in pregnancy but the term 'mother' was the preferred choice of the health professionals. Medical staff were more likely to choose 'patient' than midwives. PMID- 15835515 TI - Writing to patients: 'putting the patient in the picture'. AB - We have explored consultant, general practitioner and patient attitudes toward the proposal that following an outpatient consultation, consultants should consider communicating directly with patients in the form of a summary letter, with a copy to the referring general practitioner or other professionals as appropriate. We conducted in-depth interviews with a purposive sample of 20 consultants, 16 patients and 12 general practitioners. The consultants and general practitioners were both involved in the care of participating patients. Patients highlighted the likely value of summary letters including, increased knowledge, improvement recall of the clinical encounter, and reassurance. Clinicians were concerned that patients would not understand letters from consultants. Additional concerns included the impact of letter on consultant general practitioner relationship and medico-legal issues. These findings reflect fundamental differences in expectations about the nature and quality of communications between doctors and their patients. PMID- 15835516 TI - Healthcare worker crisis in Africa jeopardises the fight against HIV/AIDS. PMID- 15835517 TI - [Avian influenza without respiratory problems. Verified infection in a Vietnamese boy brings up a question about symptoms]. PMID- 15835518 TI - [New knowledge of inheritance and heredity and new aspects of heart disease]. PMID- 15835519 TI - [Transcranial Doppler reflects intracranial pressure]. PMID- 15835520 TI - [Hereditary cardiomyopathies: a review. Mutation of structural proteins a common cause of hereditary cardiomyopathy]. AB - Cardiomyopathy is a disorder of the cardiac muscle and can be either primary or secondary. The primary disorders have been classified by WHO into 4 groups based on structure and function; hypertrophic, dilated and restricted cardiomyopathies and arrythmogenic right ventricle dysplasia. During the last decade the familial nature of many of these cardiomyopathies has been elucidated and different genes have been found to be mutated and causative of disease. Certain patterns can be distinguished in the mutated genes, e.g. in general the genes causing hypertrophic cardiomyopathies code for proteins involved in the contractile apparatus, the sarcomere, and the genes causing dilated cardiomyopathy code for proteins that anchor the sarcomere to the cell membrane and extracellular matrix. This article reviews these recent genetic findings and discusses their potential clinical applicability. PMID- 15835521 TI - [Testicular torsion--a diagnostic dilemma. Diagnosis confirmed by exploration in only 9 per cent of cases, color ultrasound more accurate]. AB - The diagnosis was confirmed in only 9% of 170 patients surgically explored for suspected testicular torsion over a period of 2 years at four Swedish hospitals. In only every second case of testicular torsion was it considered meaningful to leave the testicle in situ. The high number of explorations, where the preoperative suspicion could not be verified, suggests that the preoperative diagnostic work-up should be improved. The use of power or colour ultrasound is suggested as a way of achieving this. PMID- 15835522 TI - [Enterocutaneous fistulas--persisting conditions but the healing capacity can be surgically restored. Current treatment and surgical skills reduce the mortality]. AB - Persisting enterocutaneous fistulae are difficult to treat. Mortality and morbidity are high. Patients are characterized by chronic infection, systemic inflammation, catabolism and impaired healing. During 1988-2002 69 patients were operated at the University Hospital, Linkoping. The mortality was 18% in 1988 to 1996 but decreased to 7% in 1997 to 2002 after establishment of present treatment strategies. It's main feature is staged surgery rising ostomies rather than doing primary anastomoses. The ostomies are closed several months later when infection and systemic inflammation are overcome and nutrition and healing capacity are restored. All patients that survived healed their fistula after surgery. Open abdomen, high output fistulas, impaired liver function and a low serum albumin were associated with a poor prognosis for healing and mortality. Effective techniques for management of abdominal defects, methods to establish enteral nutrition, minimize chronic inflammation and liver impairment as well as operative techniques and strategies are important. PMID- 15835523 TI - [Immune defense is both stimulated and inhibited by physical activity]. AB - Physical exercise may enhance some and depress other immune functions. The biological importance of these changes is not fully elucidated. Acute endurance exercise results in a relatively large redistribution of leukocytes between circulating blood and other tissues, as well as an increase in circulating cytokines. Some of these changes have been related to energy metabolism. A temporal correlation has been observed between altered immune functions and resistance to infections. A post-exercise infection can be either the result of a pre-exercise, sub-clinical infection amplified by the performed work or a novel infection, acquired during a period of decreased immune function shortly after exercise. Animal experiments have demonstrated that the susceptibility to infections after exercise depends on exercise intensity and duration, type of pathogen and time of inoculation. Exercise before inoculation with some bacterial agents can enhance resistance to infection, while exercise during an ongoing viral or bacterial infection worsens symptoms and enhances the risk for complications. Most studies demonstrate a deleterious effect of physical exercise in conjunction with infectious episodes. PMID- 15835524 TI - [The future is already here. Don't cut resources for health care and education! That's where the jobs are]. PMID- 15835525 TI - [Drug committees should be abolished because the reality has changed]. PMID- 15835526 TI - [Abortions--do Swedes always know best?]. PMID- 15835527 TI - [Special schools should teach about parenthood]. PMID- 15835528 TI - [30 out of 657 forensic psychiatric patients were diagnosed with mild mental retardation]. PMID- 15835529 TI - [Alleged overdiagnosing of breast cancer]. PMID- 15835530 TI - [Approved indication for Ezetrol]. PMID- 15835531 TI - [Care or punishment?]. PMID- 15835532 TI - [Electronic manuscripts: the periodicals earn money, the referees pay]. PMID- 15835533 TI - [Unfortunate title about fracture risk]. PMID- 15835534 TI - [Effect of long-term hypergravitation on the skeletal-muscular tissue in rats]. AB - The space flight or simulated gravitational unloading lead to the muscle atrophy, slow-to-fast transformation of muscle fibers and myofibrillar damages both in humans and animals (1, 7, 13, 17). This process could be prevented by the exercise training during space flight (1), (partly) by periodic weight support during unloading (13). It has been demonstrated in these studies that there is some level of force production necessary for the maintenance of skeletal muscle properties. It is known that adaptation to the physical training frequently induces augmentation in cross-sectional area (CSA) of muscle fibers (MF), transformation of fibers, augmentation of mitochondrial volume density, and increase in absolute volume of myofibrillas. Numerous observations suggest importance of gravitational loading in regulating muscle mass. The centrifuging is believed to be useful for preventing muscle functional and structural losses under microgravity. But there are few studies designed to investigate effect of artificial gravity on the skeletal musculature (2, 7). Our objective was to investigate structural adaptation in slow-twitch soleus muscle (percentage of connective tissue and central nuclei, fiber size, myosin heavy chain isotope, myofibrillar proteins and mitochondria volume density) after 19 and 33 days of hypergravity. PMID- 15835535 TI - [Dependence of long-lasting effects of the ACTH(4-10) analogue semax on the time of its neonatal administration]. AB - Long-lasting behavioural effects of chronic administration of synthetic ACT(4-10) analogue Semax (MEHFPGP) during early neonatal life were studied. The peptide was injected daily intraperitoneally in dose 0.05 mg/kg during the first, second or second-third weeks of postnatal development. It was shown that the peptide injections during the first week lead to a decrease and during second or second third weeks--to an increase of exploratory activity in 4-8-week aged rats. Furthermore, the peptide adminictration at all times diminished anxiety and improved learning ability of adult rats. The data obtained show that Semax neonatal administration during the first three weeks of life modulates development of brain structures involved in regulation of exploration, anxiety and learning. PMID- 15835536 TI - [Decrease of nitric oxide (NO)-cGMP-dependent vasodilatation in the vessels of lesser circulation in endothelial dysfunction]. AB - Inducible NO-synthase inhibitor aminoguanidine (AG) was used for investigation into enhanced nitric oxide (NO) production influence on elevated pressure in the pulmonary circulation (pulmonary hypertension, PH) under endothelial dysfunction. PH was simulated by subcutaneous injection of 60 mg/kg MCT to Wistar rats. Experimental groups were given AG in drinking water (15 mg/(kg x day)), and control groups were given drinking water. Rate of nitrite/nitrate excretion (RENOx) with urine was measured. The RENOx was elevated since second week as long as through the PH development. Chronic AG administration led to RENOx and soluble guanylate cyclase (sGC) NO-dependent activity restoration, and also it led to partial restoration of the right ventricular pressure. AG administration restored the perfusion pressure responses of isolated pulmonary arteries to acetylcholine. These results suggest that chronic inducible NO-synthase inhibition restores the impaired endothelium-dependent and sGC-dependent relaxation of pulmonary artery in MC-induced PH. PMID- 15835537 TI - [Seasonal changes of the activity of calcium-transporting systems in the heart and skeletal muscles of hibernating animals]. AB - In a short review of literature and according to our own data the molecular mechanisms providing seasonal adjustments of the activity of membrane Ca transporting systems (Ca-channels and Ca-pumps) in muscle tissues of animals which spent winter time in a deep torpor state (hibernation), were analyzed. Special attention was paid to changes of protein composition of sarcoplasmic reticulum membranes--the key intracellular structure responsible for excitation contraction coupling in different types of muscles, as well as to participation of endogenous protein kinases in regulation of functional activity of plasma membrane Ca-channels and sarcoplasmic reticulum Ca-ATPase during hibernation. PMID- 15835538 TI - [Acute antenatal hypoxia on different stages of embryogenesis changes the offspring behavior patterns and biogenic amine levels]. AB - The effects of acute hypobaric hypoxia on the stages of progestation or beginning of organogenesis on biogenic amines levels in brain stem and cerebral cortex of their mature offspring as well as on their behavior, were investigated. It was shown that acute hypoxia applied in the period of embryonic organogenesis resulted in severe delayed changes in offspring spontaneous behavior and led to marked changes of biogenic amine levels, particularly expressed for dopaminergic system. Females subjected to antenatal hypoxia happened to be more sensitive to its influences than males. PMID- 15835539 TI - [Na,K-ATPase and cardiac glycosides: new functions of the known protein]. AB - Review of hormonal function of endogenous cardiac steroids. Special attention is paid to recently discovered mechanism of signal transduction from Na,K-ATPase that is due to not a change of ionic gradients but to ouabain-induced alteration of enzyme conformation, that, in turn, results in interaction of the enzyme with intracellular proteins. The data concerning discovery and identification of endogenous cardiac steroids and different isoforms of Na,K-ATPase that have various sensitivity to cardiac steroid, are also considered. PMID- 15835540 TI - [Consequences of the saphenous artery denervation under normal and chronically lowered blood pressure in rats]. AB - We studied constrictor responses of saphenous artery after sympathetic denervation in normotensive rats and rats with chronic regional hypotension. Abdominal aorta was partially occluded in Wistar rats distally to the renal arteries, lowering blood pressure in the hindquarters by about 40%, a week later to denervate saphenous artery the femoral nerve was cut. The density of periarterial nerve plexus and neurogenic responses of the vessel restored partially in 2 weeks and completely in 6 weeks after the surgery; the chronic hypotension did not modify the dynamics of reinnervation. Arteries of both groups of rats demonstrated higher sensitivity to noradrenaline during 6 weeks after denervation, whereas vessel sensitivity to serotonin was enhanced only in normotensive rats. Therefore, chronic hypotension may prevent postdenervation hypersensitivity of vascular smooth muscle to vasoconstrictors. PMID- 15835541 TI - [Effect of new synthetic anxiolytic selank on gastric wall blood flow and mesenteryc lymphatic vessels contractility in anesthetized rats]. AB - The influence of a new heptapeptide Selank on microcirculation in anesthetized white rats was investigated. Selank (Thr-Lys-Pro-Arg-Pro-Gly-Pro) is a new synthetic anxiolytic which exerts obvious antiulcerogenic action and promotes healing of experimental ulcers. Action of the peptide on gastric blood flow in the stomach by using the method ofhydrogenic clearance and Selank action on mesenteric lymphatic contractility by microscopical observation in situ, were studied. Selank fail to influence basal gastric blood flow but it can normalize blood flow reduced by indomethacin. The study of dose-depended of Selank effect on lymphatic vessels contractility showed that its low concentration (10(-12)- 10(-14) M) enhanced amplitude and increase frequency of lymphatic vessel contractions which indicates an enhancement of lymphatic flow. The high doses of peptide (10(-6)--10(-10) M) also augmented the contraction amplitude but decreased its frequency. The maintenance of adequate blood flow and lymphatic vessel contractility can be one of the mechanisms of the Selank antiulcerogenic properties. PMID- 15835542 TI - [Mechanisms of discrimation of the prolatin physiologic action on the liver at the receptor level of hepatocytes and cholangiocytes]. AB - The role ofprolactin receptor isoforms in discrimination ofprolactin effects on liver is summarized. The necessity of studying of parameters ofprolactin receptor expression in differentially isolated different types of liver cells is demonstrated. The review is illustrated by data on different regulation by sex hormones and obstructive cholestasis of expression of prolactin receptor isoforms in rat hepatocytes and epithelial cells of bile ducts. The mathematical model is introduced for estimation of intensity of prolactin-induced signal cascades on the basis of experimentally measured parameters of prolactin receptor expression. Some further steps of the investigation of prolactin effects discrimination at the level receptor unit are suggested. PMID- 15835543 TI - [Effect of the medicinal leech salivary gland secretion on the state of rat subcutaneous mast cells]. AB - It is firstly showed that the medicinal leech salivary gland secretion (SGS) as a polycomponent system of proteins and low-molecular weight substances, activates rat subcutaneous mast cells in vitro prompting a decrease in the heparin saturation index and increasing some characteristic mast cells morphometric parameters. The same mast cell changes were detected by analysis of some specimens of subcutaneous cellular tissue in the point of skin injured by the leech bite. It is shown that these changes are saved during 3 days. The mechanical injury of rat skin does not effect the mast cells activation. Activation of mast cells by SGS is extended to the distant subcutaneous mast cells. It is expressed in sharp decreasing of heparin saturation index although not statistically positive. The secondary leeching on these distant points provokes reduction of mast cells activation and some decrease of post-leeching blood heparin content: 0.154 +/- 0.03 units/ml (n = 10) as compared with post leeching blood heparin contents analysed from the wound after the primary leeching (0.160 +/- 0.03 units/ml, n = 10). Proceeding from these findings, participation of heparin secreted from activated mast cells in the support of post-leeching bleeding is suggested, the phenomenon which provides unloading of capillary pool by application of medicinal leeches for treatment many diseases. PMID- 15835544 TI - [Nikolai Ignat'evich Bakst--one of I. P. Pavlov's university tutors]. PMID- 15835545 TI - Encouraging workers to save: the 2005 Retirement Confidence Survey. AB - This Issue Brief reports findings of the 15th annual Retirement Confidence Survey (RCS), which points to potential solutions to the American retirement savings problem, specifically ways that could help workers save more through their employment-based retirement plans. IMPORTANCE OF EMPLOYER MATCH: More than 7 in 10 workers not currently contributing to their employer-sponsored retirement plan say an employer contribution of up to 5 percent of their salary would make them much more or somewhat more likely to participate (72 percent). SIMPLIFIED OPTIONS: Other retirement plan options that nonparticipants say would make them more likely to contribute are an investment option that automatically becomes more conservative as their retirement date approaches (66 percent) and a feature that automatically raises workers' contributions by a fixed amount or percentage when they receive a pay raise (55 percent). Two-thirds of nonparticipants indicate they would be very or somewhat likely to remain in their employer's plan if they were automatically enrolled (66 percent). SOCIAL SECURITY: Nearly 7 in 10 of today's workers are skeptical that Social Security will continue to provide benefits of at least equal value to those received by current retirees (68 percent). This proportion has remained relatively constant in recent years, but is below the 1995 level (79 percent). Workers continue to be unable to identify the age at which they will be eligible for full Social Security benefits. MOST BEHIND SCHEDULE IN SAVING: A majority of workers believe they are behind schedule when it comes to planning and saving for retirement (55 percent). Most of those behind schedule say that high expenses, particularly everyday expenses (49 percent), child-rearing expenses (39 percent), and medical costs (35 percent), are a major factor in keeping them from saving. LESS THAN HALF HAVE TRIED TO CALCULATE NEEDED SAVINGS: Approximately 4 in 10 workers say they have tried to calculate how much they need to accumulate for retirement. More than one-third of these workers say they asked a financial advisor to calculate this number or used their own estimates; 10 percent say they simply guessed how much they will need in retirement. PMID- 15835546 TI - Did he really say that? PMID- 15835547 TI - Magazine cover story good news and bad news. PMID- 15835548 TI - Consider profit margin of NP employers. PMID- 15835549 TI - Cash-based care. PMID- 15835550 TI - Questions about certification. PMID- 15835551 TI - Ganglion cyst aspiration. PMID- 15835552 TI - Patient information. Understanding urinary incontinence. PMID- 15835553 TI - Andropause and the aging man: separating evidence from speculation. PMID- 15835554 TI - Osteoporosis in men. PMID- 15835555 TI - Umbilical cord blood banking. PMID- 15835556 TI - Detection and diagnosis of scrotal masses. PMID- 15835557 TI - Circumcision of infant boys: parents require education to make informed decision. PMID- 15835558 TI - Meeting the needs of severely obese patients. PMID- 15835559 TI - HIV takes toll on ethnic and sexual minorities. PMID- 15835560 TI - The quest for a little pink pill: female sexual dysfunction finally attracting attention. PMID- 15835561 TI - Animal cruelty as a predictor of human violence. PMID- 15835562 TI - Pain and laughter in the ED. PMID- 15835564 TI - [Study about the benefits of vestibular rehabilitation in peripheral vestibular disorders]. AB - BACKGROUND: Vestibular rehabilitation in vestibular disorders. AIM: To study the benefits of Vestibular Rehabilitation through the Brazilian Dizziness Handicap Inventory-DHI. METHOD: Application of the Vestibular Rehabilitation proposed by Zee (1985) and analysis using the Sign; T-Student and Kendall's T tests. RESULTS: A change in the diagnosis of vestibular peripheral disorder to normal vestibular patterns was obtained in 75% of the cases. CONCLUSION: Our results indicated a change of the diagnosis and in the quality of life of the tested individuals. The use of the DHI demonstrated to be an excellent tool for qualitative studies. PMID- 15835565 TI - [Simplified auditory processing test and difficulties in reading and spelling]. AB - BACKGROUND: Auditory processing disorders and difficulties in reading and spelling. AIM: To analyze the answers presented by children with reading and spelling disorders in a simplified auditory processing test and in a four sequential syllables memory test. METHOD: Participants of this study were 9 children who attended a special program at school, with no hearing impairments and with no history of neurologic and/or psyquiatric disorders. RESULTS: A significant relation was found between reading and spelling difficulties, and failing in the four sequential syllable memory test and instrumental sounds tasks, demonstrating that these tasks were efficient in detecting this type of difficulties. CONCLUSION: A relation exists between auditory processing and reading and spelling difficulties. PMID- 15835566 TI - [Speech perception in cochlear implant users: two programming situations]. AB - BACKGROUND: In order to shorten the programming process of cochlear implanted (CI) children it is possible to establish the T level (threshold level) as being 10% of the C level (most comfortable level). AIM: To compare the results of speech perception tests using the map of the real thresholds levels and the map corresponding to 10% of the C level. MRTHODS: A list of monosyllables and sentences were used with 30 CI individuals in two programming situations. RESULTS: no difference was found between the results in the speech tests in the two programming situations. CONCLUSION: The use of the map with the T level corresponding to 10% of the C level can be an option to program young children, especially for those who do not respond to the investigation of the real T levels. PMID- 15835567 TI - [Sequential verbal memory for three and four syllables in school children]. AB - BACKGROUND: Auditory memory ability. AIM: To characterize the auditory memory ability for verbal sequences in children with ages ranging from 3 to 12 years. METHOD: A memory test for verbal sequences was used, as well as a variation with the addition of one syllable to each sequence. Participants were 223 students of public and private schools. RESULTS: All subjects repeated at least two out of the three sequences of three syllables. At the age of nine, they were capable of repeating all of the three sequences. At the age of six, they repeated at least two out of the three sequences of four syllables. CONCLUSION: The behavioral responses to verbal sounds presented in sequences, improves as children grow older. Children of private schools demonstrated an earlier development of this ability. PMID- 15835568 TI - [Effect of the early intervention of oral habits on the development of dental occlusion]. AB - BACKGROUND: Sucking habits are related to malocclusion. AIM: To evaluate the effect of a counseling interview about inhibiting this oral habit and, its influence on dental occlusion and labial-lingual posture. METHOD: 40 children (mean age = three years), with a sucking habit and anterior open bite, received counseling aiming for the interruption of this habit. Measurements of the open bite were made, as well as a myofunctional evaluation, prior to counseling and, three and six months after it. RESULTS: 26 children stopped with the habit of sucking, with 25 children presenting a reduction of the open bite. No change was observed in labial-lingual posture. CONCLUSION: The counseling interview favored the interruption of the oral habit and the correction of the open bite. PMID- 15835569 TI - [Relation between phonologic and pragmatic habilities in specific language impairment]. AB - BACKGROUND: Specific language impairment. AIM: To establish relations between the performance of subjects in phonologic and pragmatic tests. METHOD: Tabulation and analyses of the data of phonologic and pragmatic tests of 20 children with ages raging from 3:0 to 5:11 years, with the diagnoses of Specific Language Impairment (SLI). RESULTS: The statistic analyses showed good correlations between developmental processes (DP) and non-developmental processes (NDP), DP and the use of verbal mean, vocal mean and number of communication acts per minute. CONCLUSION: The performance in the phonological and pragmatic tests does not seem to have a direct correlation in subjects with specific language impairment. PMID- 15835570 TI - [Assessment of word finding skills in the discourse of adolescents]. AB - BACKGROUND: Relation between the abilities of word finding and reading. AIM: Application of a word finding test to the discourse of two groups of adolescents which differed in their abilities to comprehend written texts. METHOD: Application of the Test of Word Finding Discourse in 20 participants (group 1 = 10 e group II = 10). RESULTS: There was no difference between the groups regarding the global word finding index; as for specific behaviors, word substitution was statistically significant and did differentiate the groups; as for the comparison with American standards, the use of unspecific words was statistically significant; the expected linear decrease for all of the behaviors was not observed. CONCLUSION: A linear and reciprocal relation between word finding and difficulties in reading does not exist. PMID- 15835571 TI - [Language therapy results with adolescents of the autistic spectrum]. AB - BACKGROUND: Results of language therapy with adolescents and children with autistic spectrum disorders. AIM: to identify changes, after a six-month period of speech-language therapy, in a group of adolescents and to compare them with a group of children. METHOD: Data referring to the communicative profile and social cognitive performance of 12 adolescents and 12 children were analyzed. RESULTS: Just one of the 10 observed areas presented meaningful progress for the group of adolescents and 5 areas presented progress for group of children. CONCLUSION: The results reinforce the importance of language therapy for autistic children, as more progress can be obtained at this age. PMID- 15835572 TI - [The role of child's play in the relationship between hearing mothers and deaf children]. AB - BACKGROUND: The importance of child's play and its assessment for the therapy of deaf children. AIM: To analyze the organization of the act of playing between hearing mothers and deaf children, how it can be assessed and its relation with language acquisition. METHOD: Analysis of video recordings of six deaf children playing with their mothers in three different moments, with intervals between the recordings of 6 months each. Criteria were established for the analysis of the data. RESULTS: All of the children presented a language acquisition delay and a very poor interaction with their mothers. During one year five out of the six mothers modified their way of playing. CONCLUSION: Playing might (and should) be an efficient parameter to assess the therapy of deaf children. PMID- 15835573 TI - [Teaching sign language to the families of the deaf: focusing the learning process]. AB - BACKGROUND: According to bilingual education, only through sign language will deaf children attain linguistic and cognitive development, enabling them to learn a second language--spoken or written. However, it is also necessary for families to learn sign language in order to have a more efficient communication. AIM: To analyze methodological aspects of the teaching-learning process of Sign Language to family groups. METHOD: Transcription and analysis of video recordings were made. RESULTS: The practice of teaching of the deaf teacher modifies itself during the research period and his attitude influences the way by which parents participate. CONCLUSION: The teaching methodology used by the deaf teacher interferes significantly in the motivation/participation of parents, followed by the acceptance of deafness and sign language. PMID- 15835574 TI - [Generalization based on implicational relations in subjects treated with phonological therapy]. AB - BACKGROUND: Generalization based on implicational relations. AIM: To analyze generalization based on implicational relations in subjects with mild-moderate phonological disorder, who underwent therapy based on the Maximal Oppositions Model. METHOD: The initial phonological system of four subjects was analyzed, taking in consideration sounds that were not completely acquired and their changes in probes. A final phonological assessment was made. RESULTS: All subjects presented generalizations based on implicational relations and expanded their phonological systems. CONCLUSION: The Maximal Oppositions Model favored generalizations based on implicational relations. PMID- 15835575 TI - [Language characterization in the X-fragile syndrome: a review study]. AB - BACKGROUND: X-Fragile Syndrome. AIM: To compile information about the language, cognitive and behavior alterations in the X-Fragile Syndrome, using the results of previously published studies and to present the standardized instruments used as testing materials. CONCLUSION: studies used formal and informal testing to assess language. The results present variability regarding the linguistic deficits, which are influenced by the level of the cognitive deficit and behavior alterations. Alterations of the oral praxes and of speech articulation are also expected. PMID- 15835576 TI - The financial return on Magnet recognition. AB - There are many ways the Education Department may bear some of the expenses of Magnet application, and they should be budgeted accordingly. However, there is a great return on the investment of educating and developing the staff, and the Education Department can share in that success. PMID- 15835577 TI - Tuberous sclerosis complex. PMID- 15835578 TI - Online learning communities can provide support for nurses preparing for certification examinations. AB - Achieving certification is a benefit to the nurses, to their patients, and to the organizations that support them. Developing an online learning community is a simple way for the institution to offer support to the nursing staff. Providing the resources for creation of online learning communities demonstrates the facility's commitment to communication, education, and professional development. PMID- 15835580 TI - Critical thinking in nursing: an integrated review. AB - Critical thinking skills are essential to function in today's complex health care environment and to ensure continuing competence for the future. This article provides a review of various definitions and research studies related to critical thinking. Educators and researchers need to clearly define critical thinking, because there has been wide variation in definitions and descriptions of critical thinking. Research studies have shown inconsistent findings, and many have used a one-group pretest-posttest design over a single course or during the length of a nursing program. Studies have not shown an association between critical thinking and competence; rigorous research studies are needed to understand the process of critical thinking. PMID- 15835581 TI - Nurturing critical thinkers. AB - The continuing education environment offers unique opportunities to assist nurse learners in developing critical thinking skills. Adjustments in typical teaching strategies and learner approaches to learning are required to facilitate this process. Suggestions are provided to assist the continuing education educator in developing, implementing, and evaluating learning activities that stimulate critical thinking abilities. PMID- 15835582 TI - Engaging the body and mind with the spirit of learning to promote critical thinking. AB - Adult learners want to know the benefits of paying attention to continuing education and how it can improve the care they provide. Linking the body and mind with the spirit of learning can improve the likelihood of meeting both the staff's needs and the educator's goals in learning. Applying these brain-based strategies can engage the learner, focus attention, and increase the involvement of the professional level staff. These can be applied in a variety of different formats. Examples are provided for self-directed learning formats, which can be a versatile and cost-effective way to communicate continuing education to the learner. PMID- 15835583 TI - Strategies for active learning in online continuing education. AB - Online continuing education and staff development is on the rise as the benefits of access, convenience, and quality learning are continuing to take shape. Strategies to enhance learning call for learner participation that is self directed and independent, thus changing the educator's role from expert to coach and facilitator. Good planning of active learning strategies promotes optimal learning whether the learning content is presented in a course or a just-in-time short module. Active learning strategies can be used to enhance online learning during all phases of the teaching-learning process and can accommodate a variety of learning styles. Feedback from peers, educators, and technology greatly influences learner satisfaction and must be harnessed to provide effective learning experiences. Outcomes of active learning can be assessed online and implemented conveniently and successfully from the initiation of the course or module planning to the end of the evaluation process. Online learning has become accessible and convenient and allows the educator to track learner participation. The future of online education will continue to grow, and using active learning strategies will ensure that quality learning will occur, appealing to a wide variety of learning needs. PMID- 15835584 TI - Peer coaching: the next step in staff development. AB - A common problem in continuing nursing education and staff development is the transfer of learning to clinical practice. Peer coaching offers a solution to this problem. Initiated by educators, peer coaching has been researched in educational settings and found to be effective in facilitating the transfer of newly acquired knowledge and skill into classroom teaching strategies. This article describes the background, components, process, characteristics, and benefits of peer coaching. A specific example of using peer coaching to teach clinical breast examination skills is used to illustrate the application of peer coaching to the staff development of healthcare professionals. Peer coaching is the next step in nursing staff development. PMID- 15835585 TI - Can acute mesenteric ischemia be ruled out using computed tomography? Critically appraised topic. PMID- 15835586 TI - Lessons from Ivan Illich. PMID- 15835587 TI - Coronary computed tomography angiography: emerging technique for coronary artery imaging--pictorial essay. PMID- 15835588 TI - Computed tomographic findings of abdominal complications of Crohn's disease- pictorial essay. PMID- 15835589 TI - Patient compliance with recommended follow-up after fine-needle aspiration biopsy of solid breast masses. AB - OBJECTIVE: Any needle biopsy procedure is associated with a false-negative rate; therefore, masses diagnosed as benign by needle biopsy should be followed to confirm their stability. This study determines patient compliance with recommended 6-month follow-up of breast masses that have had benign cytology. MATERIALS AND METHODS: This study consists of a retrospective 5-year chart review from August 1994 to August 1999. The study includes patients who had an ultrasound-guided fine-needle aspiration biopsy of a solid breast mass diagnosed as benign and were told to return in 6 months for follow-up. Patients who had the masses surgically removed were excluded. Patients who did not follow-up within a 1-year period were sent a letter of reminder. RESULTS: Over the 5-year period, a total of 462 patients had an ultrasound-guided fine-needle aspiration biopsy of a solid breast mass diagnosed as benign and were instructed to return at 6 months for follow-up. Two hundred ninety-six (64%) of these patients returned for follow up within 1 year. The remaining 166 patients were sent a letter of reminder. Of these 166 patients, 71 (43%) responded to the letter; 5 patients were lost to follow-up because they had moved. Of all 462 patients, 367 (79%) eventually returned for follow-up, at an average of 9.2 months after biopsy. CONCLUSIONS: Over a 5-year period, 296 out of 462 (64%) patients were compliant with recommendations for follow-up, and an additional 71 patients responded to a letter of reminder. Noncompliance with recommended follow-up could lead to a delay in diagnosis of false-negative lesions. This study demonstrates the need for radiologists to have a system to track compliance. PMID- 15835590 TI - Externally placed vs intravaginally positioned radio frequency coils for quantitative spin-spin relaxometry of ovarian follicular fluid. AB - OBJECTIVE: To evaluate different imaging protocols, especially with respect to radio frequency (RF) receiver coil location, for their suitability in providing least squares derived quantitative T2 values of ovarian follicular fluid for investigations of basic ovarian physiology. METHODS: The ovaries of 10 women were imaged via magnetic resonance imaging (MRI) using externally positioned and intravaginally placed RF receiver coils. Half-Fourier acquisition with single shot turbo spin-echo (HASTE), multiple-echo T2, Dixon, turbo spin-echo, and 3 dimensional (3D) fast imaging with steady-state precession (FISP) and time reversed FISP (PSIF) sequences were used. Quantitative T2 nuclear spin relaxation rate information from the ovarian follicles between data acquired with the external and intravaginal coils were compared. Additionally, the amount of ovarian follicle and corpora lutea structural detail visible was qualitatively assessed. RESULTS: The T2 computations indicated that there was no difference in the follicular fluid T2 values or in the heterogeneity (spatial variance) of the T2 values between data acquired with the external RF coil and data acquired with the intravaginal RF coil. The best sequences for the visualization of ovarian internal structure were the 3D PSIF sequences and the multiple-echo T2-weighted images, confirming our earlier imaging work on excised cow ovaries. CONCLUSION: It is best to use an externally placed RF coil for quantitative MRI study of ovarian physiology given the lack of difference in quantitative T2 information and the difficulty associated with imaging the ovaries using an intravaginal RF probe. PMID- 15835593 TI - The winking happy face. PMID- 15835594 TI - The risks of an ownership society. PMID- 15835595 TI - The failed conversion of CareFirst BlueCross BlueShield to for-profit status: Part 2, Lessons learned. AB - This article presents lessons for other states and all types of nonprofit health care organizations from the failure by nonprofit CareFirst Blue Cross Blue Shield to convert to for-profit status and be sold to the publicly traded Wellpoint Health Networks, Inc. The lessons relate only in part to conversions. More broadly they concern any kind of strategic decision making by nonprofit health care boards of directors and their executives that substantially affects the public interest. This article is a companion to one by this same author published in the Fall 2004 issue of Inquiry. That article chronicled the events and political environment surrounding the conversion proposal, the review process and decision, and the aftermath of actions and reactions by various parties, including state legislation to clarify CareFirst's mission and to reform its board. PMID- 15835596 TI - Doing away with uncompensated care of the uninsured. AB - As more people lose their health insurance, an increasing volume of uncompensated care is absorbing billions of dollars of limited resources of the nation's hospitals, health systems, and other provider organizations. As yet, no provider organization has developed a comprehensive management approach to address this growing problem. Currently, the money spent on uncompensated care is viewed as a drain on institutional bottom lines rather than as a fund dedicated to improving the health of uninsured patients and prospective patients. If an accountable executive were made responsible for managing this problem by paying for all of this care on a case-by-case basis as third parties do, uncompensated care could be eliminated. Payment for each case would come from the institutional resources no longer required for uncompensated care. There is reason to hypothesize that with effective management, a significant amount of the resources currently absorbed by uncompensated care could be shifted from excessive inpatient care to more productive, innovative community initiatives. This paper outlines a six point management program designed to increase the income and decrease the expense currently associated with uncompensated care, while improving quality, patient satisfaction, and outcomes. The program can be carried out by an individual provider organization, or as a collaborative program involving two or more organizations. Recommended are demonstration projects to test the feasibility and net cost or cost savings of such an approach, preferably starting in one-hospital towns. PMID- 15835597 TI - The managed care backlash: did consumers vote with their feet? AB - The managed care backlash led many to predict the demise of health maintenance organizations (HMOs). This paper examines trends in HMO enrollment in all metropolitan communities from 1994 to 2000 to identify factors that led to diminishing enrollment in the backlash era and circumstances in which HMOs maintained or expanded their presence. We use a database constructed from a wide variety of sources that describe HMO penetration and other characteristics of all metropolitan statistical areas. We found the backlash is not evidenced in a large degree of consumer switching. However, HMOs were more likely to maintain their presence in areas with high-cost growth and with greater managed care experience. Medicaid HMO growth continued to expand rapidly, indicating the possibility of a two-tiered system in which low-income beneficiaries have less choice than the privately insured. PMID- 15835598 TI - State payment limitations on Medicare cost-sharing: impact on dually eligible beneficiaries. AB - The Balanced Budget Act (BBA) of 1997 allowed states to limit how much their Medicaid programs contributed toward the Medicare cost-sharing liability of dually eligible beneficiaries. Policymakers have grown concerned that such limitations may affect access to care for these beneficiaries. We used a quasi experimental design to analyze changes in access from 1996 to 1998, using Medicare and Medicaid data from nine states. Cost-sharing payments fell in six of the nine states following the BBA, and access to outpatient physician visits for dually eligible beneficiaries was reduced relative to non-dually eligible beneficiaries in those states. PMID- 15835599 TI - The Balanced Budget Act of 1997 and U.S. hospital operations. AB - The Balanced Budget Act (BBA) of 1997 initiated several changes to Medicare payment policy in an effort to slow the growth of hospital Medicare payments and ensure the future of the Medicare Hospital Insurance Trust Fund. Although subsequent federal legislation relaxed some original proposals, restored funds were limited and directed to specific types of hospitals. In addition, these Medicare policy changes came at a time when hospitals faced private sector payment constraints. This paper assesses the short-term effects of the BBA on operations of nonprofit hospitals in the United States and compares these effects to those observed in the early 1980s during implementation of the Medicare prospective payment system (PPS). We found that some operational changes instituted by hospitals facing financial pressures from the BBA were similar to those observed for hospitals that faced pressure from Medicare PPS, including efforts to contain Medicare cost growth, to expand outpatient service provision, and to contain hospital staffing. However, during PPS implementation hospitals experienced declining inpatient use and growing profit margins, whereas post-BBA hospitals experienced growing inpatient use and declining margins. PMID- 15835600 TI - The effect of Medicare's prospective payment system on discharge outcomes of skilled nursing facility residents. AB - In July 1998, the Centers for Medicare and Medicaid Services (CMS) changed the payment method for Medicare (Part A) skilled nursing facility (SNF) care from a cost-based system to a prospective payment system (PPS). Unlike the previous cost based payment system, PPS restricts skilled nursing facility payment to pre determined levels. CMS also reduced the total payments to SNFs coincident with PPS implementation. These changes might reduce quality of care at skilled nursing facilities and could be reflected in resident discharge patterns. The present study examines the effect of the 1998 policy change on resident discharge outcomes. The results indicate that PPS reduced the relative risk of discharge to home and to death for Medicare residents (compared to non-Medicare residents) and had no significant effect on hospitalizations or transfers. PMID- 15835601 TI - Quality report cards, selection of cardiac surgeons, and racial disparities: a study of the publication of the New York State Cardiac Surgery Reports. AB - Quality report cards have become common in many health care markets. This study evaluates their effectiveness by examining the impact of the New York State (NYS) Cardiac Surgery Reports on selection of cardiac surgeons. The analyses compares selection of surgeons in 1991 (pre-report publication) and 1992 (post-report publication). We find that the information about a surgeon's quality published in the reports influences selection directly and diminishes the importance of surgeon experience and price as signals for quality. Furthermore, selection of surgeons for black patients is as sensitive to the published information as is the selection for white patients. PMID- 15835602 TI - Health plan disenrollment in a choice-based Medicaid managed care program. AB - Consumer decisions to switch health plans have implications for quality of care and risk selection. We examine factors related to time to disenrollment in a Medicaid managed care program where beneficiaries face a menu of plans and can change plans every month. Several findings have direct policy relevance. Families and individuals who make active choices upon entering the program are at substantially lower risk of disenrollment than those who are auto-assigned. Interactions between enrollee ethnicity and provider language proficiency suggest that enrollee satisfaction depends on the cultural competence of providers. Differential disenrollment by risk status results in adverse retention for certain types of plans. PMID- 15835603 TI - Electronic prescribing and HIPAA privacy regulation. AB - Electronic prescribing offers the prospect of safer medication management, but fulfillment of that promise depends on ready access to personal health information from many sources, thus raising new concerns about information privacy and security. Federal privacy regulations under the Health Insurance Portability and Accountability Act of 1996 (HIPAA) limit the sharing of health information by providers, and particularly may discourage information sharing over distributed computer networks. This analysis finds that although HIPAA has only a limited effect on current e-prescribing practices, future electronic prescribing systems will likely fall short of their potential benefits, absent policy refinements designed to encourage clinically appropriate, networked sharing of patient health information. PMID- 15835604 TI - Are viable Mycobacterium leprae present in lepromatous patients after completion of 12 months' and 24 months' multi-drug therapy? AB - A study was carried out to determine whether or not viable bacilli persist in MB patients treated with 12-month and 24-month multidrug therapy (MDT). In the first group, 60 untreated lepromatous patients who had an initial average bacterial index (BI) of 3+ or more were enrolled. At the completion of 12 months of MDT, skin biopsies were obtained and M. leprae concentrate was inoculated into the footpads of five thymectomized and irradiated (T900r) mice. Rees technique was used for the mouse footpad (MFP) experiment. Harvesting was done it the 6th, 9th and 12th months. Out of the 60 biopsies inoculated into mouse footpads to check the viability of bacilli, 2 skin biopsies (3.3%) showed significant growth and 10 (16%) showed equivocal growth. 27 patients also had nerve biopsies tested for growth in MFP studies. None of the inoculated nerve biopsies showed significant multiplication in the MFP experiments. However, 4 biopsies (14%) showed equivocal growth. In the second group, 20 patients had skin biopsies and 10 had nerve biopsies done at the end of 24 doses of MDT in order to test the viability of bacilli; none of the skin or nerve biopsies from these patients showed any growth. This study showed that M. leprae present in the tissues after 24 doses of MDT are not viable and the drug schedule of 24 doses is adequate to treat leprosy patients, irrespective of their BI. However, a small (3.3%) percentage of the patients with a high BI harbour viable bacteria in the skin after 12 doses of treatment. Since a large majority of the patients (38 patients) who had a high initial BI responded well to the treatment, it is important to find out the reason for the lack of response in two patients. One of the reasons may be the presence of drug-resistant strains. It is important to follow up on these patients for a longer duration to ascertain whether or not they would relapse. PMID- 15835605 TI - Lasso procedure to correct finger-clawing in leprosy using index or middle finger flexor digitorum superficialis split into four tails--a long-term follow-up study. AB - For obvious reasons, the use of flexor digitorum superficialis (FDS) from the ring finger, for correction of finger-clawing, is usually not recommended in leprosy. Hence, one has to choose either index or middle finger FDS for correction of finger-clawing. No significant differences could be made out when follow-up data of claw-finger correction by modified lasso procedure, using FDS either from index or middle finger, were compared. In some hands post-operative problems, such as stiffness, superficialis minus deformities of proximal interphalangeal joints (PIPJ) and distal interphalangeal joints (DIPJ), were noted. As revealed by finger dynamography, the working space of the hand was not found to be fully restored, the donor finger showing distortion of its working space. PMID- 15835606 TI - Incidence of neuritis among paucibacillary leprosy patients during treatment and surveillance. AB - A cohort study was done on 180 patients receiving the standard WHO PB-MDT regimen at the TLM Hospital in Kolkata, during MDT and 2 years of surveillance to determine the incidence of neuritis. Neuritis occurred in only 2 out of the 180 patients, confirming that the current WHO treatment for PB-MDT is attendant with minimal risk of neuritis. No risk factors were found, except pregnancy in a female patient. While addition of clofazimine in the new U-MDT under trial might take care of the occasional neuritis, further clinical research might be useful to identify sub-groups of PB patients at risk of neuritis. PMID- 15835607 TI - Sub-polar lepromatous leprosy presenting as urticarial wheals: a case report. AB - A 40-year male lepromatous leprosy patient presented with four, slightly erythematous patches, with multiple urticaria-like wheals, distributed over the body symmetrically and with thickened right ulnar and right common peroneal nerves. He was previously diagnosed as a case of chronic idiopathic urticaria. Slit skin smear from the patches and urticarial wheals showed a BI of 3+ and skin biopsy showed some features of borderline lepromatous leprosy (BL). PMID- 15835608 TI - Rate of relapse and reactions in MB leprosy patients after 24 and 12 months of MDT in Maharashtra. PMID- 15835609 TI - Scrotal sinuses in histoid leprosy. PMID- 15835610 TI - Corrective surgery for deformities in women leprosy patients. PMID- 15835611 TI - Summary report of the 23rd Biennial Conference of the Indian Association of Leprologists. Haldia, West Bengal, 27-29 February 2004. PMID- 15835612 TI - Professional profile of radiologic technology educators. AB - OBJECTIVE: Full-time radiologic technology educators (n = 565) were surveyed to determine their demographic characteristics and professional profile. RESULTS: Overall, the majority of radiologic technology educators surveyed were women between the ages of 40 and 59, had a bachelor's or master's degree, were certified in radiography and reported annual incomes from 40,001 dollars to 60,000 dollars. Most educators spent between 1 hour and 8 hours per week on classroom instruction/laboratory and in the clinical setting. Additionally, hospital or community college programs employed the majority of educators. Demographic characteristics of radiologic technology educators varied according to the type of institution in which they were employed and by education level. CONCLUSION: Study findings show a potential loss of qualified educators in the near future and the need for increased efforts to prepare and recruit radiologic technologists into the education career path. PMID- 15835613 TI - Acceptance of distance learning by radiologic technologists. AB - OBJECTIVE: A national survey of 1300 registered radiologic technologists was conducted to determine acceptance of distance learning for continuing education, advanced certification and higher academic degree requirements. MAIN OUTCOME MEASURES: Differences between the demographic data and reported acceptance levels were compared using the chi2 (chi-square) test of significance. RESULTS: None of the demographic variables indicated statistically significant differences in acceptance levels. T tests were performed for continuous variables with no significant differences. A rank-order analysis was performed to determine the most preferred method of distance learning. Internet, correspondence and video tape courses were reported to be the most preferred methods for distance learning. CONCLUSION: Study results suggest a high level of acceptance of distance learning for continuing education (95%), advanced certification (93%) and higher academic degree requirements (87%). PMID- 15835614 TI - Systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease that exhibits a wide range of clinical symptoms, including skin rashes, oral ulcers, arthritis, pulmonary pleuritis, anemia and seizures. Medical imaging plays a role in assessing the extent of disease, neurological involvement, treatment complications and disease progression. This article describes the known risk factors for SLE and SLE diagnosis and treatment. PMID- 15835615 TI - The evolution of PET-CT. AB - Positron emission tomography-computed tomography (PET-CT) was the first fused or combined medical imaging technique. Although PET-CT has received widespread acclaim as a major imaging advancement, many questions have surfaced regarding its use. This article answers some of these questions and examines what PET-CT means to medicine and the medical imaging community. PMID- 15835616 TI - Modified problem-based learning. PMID- 15835617 TI - Candid camera. PMID- 15835618 TI - The 411 on molecular imaging. PMID- 15835619 TI - Patient page. MR imaging of the knee. PMID- 15835620 TI - [A malignant tumour in the breast is not always primary breast cancer]. AB - In a 75-year-old woman with a swelling in her left breast, a 39-year-old woman with an anal fissure due to diarrhoea and a 65-year-old woman with chest pain, a mammary tumour was diagnosed that did not originate in mammary tissue. These were a recurrent melanoma, a carcinoma of the thyroid and a B-cell lymphoma, respectively. All patients were treated. The first patient developed new metastases one year later, the second died, partly as a result of the tumour, and the third showed no recurrence of the tumour after two years. Breast cancer is one of the most frequently occurring neoplasms in women. Primary tumours in the breast from other origins and metastatic lesions to the breast from extramammary tumours are rare. Most of these cases concern haematological malignancies and metastases from melanoma and lung cancer. Despite the fact that metastases to the breast are rare, one should always consider the possibility. PMID- 15835621 TI - [Behaviour that exceeds the limits]. AB - When medical students first enter the clinic, they will experience situations in which they feel threatened or intimidated. In some cases, this may entail sexual harassment, as described by Van den Muijsenbergh and Lagro-Janssen in their study of students at Nijmegen Medical School. Such behaviour is often difficult to prevent if it concerns patients, especially psychiatric patients. However, it should be prevented if it concerns staff or supervisors. In all cases, students should be aware that such incidents may happen and be prepared to deal with them. They also deserve a safe environment in which to share and discuss such experiences. It is up to the University Medical Centres to provide such a safe haven. PMID- 15835622 TI - [Less referrals from general pratitioners to medical specialists: the desired results of a consistent policy]. AB - In a comparison of the 1987 and 2001 Dutch national studies of diagnoses and performance in general practice, a decline was observed in referrals to pediatricians. This decline concerned all pediatric subspecialties, but was particularly marked for acute otitis media and visual impairment. This trend is in line with other observations of general practitioners' interventions in the last two decades. General practitioners treated a larger proportion of health problems presented to them in 2000 (96% of the episodes) than in 1987 (91%). A decline in referrals of all age groups and all specialties was also reported for 1971--1994. The decline in referrals took place in an aging population, while the annual number of contacts between patients and their general practitioners did not change substantially. This reflects the fact that since 1970 health care policy has been directed at a strengthening of general practice. The decline in referrals proves the success of this policy. PMID- 15835623 TI - [Report from the Health Council of the Netherlands on the chronic fatigue syndrome: moving away from the body-mind dichotomy with a view to effective prevention and treatment]. AB - The Health Council of the Netherlands has issued a report on the chronic fatigue syndrome (CFS). CFS is a real and seriously debilitating condition which imposes limitations on an individual's personal, occupational and social functioning. It is a syndrome of unknown aetiology without physical signs or biological markers. Although there is no disease, patients both feel ill and give the appearance of being ill. There is no consensus on whether CSF patients are able to work or whether they should be entitled to social security benefits. An imbalance between demand and coping is central in CFS, with stress as an important intermediary factor. It is little use concluding that unexplained signs are 'psychological' or that 'I cannot find anything wrong with you so you must be healthy'. The classical view that mind and body are separate systems is outmoded. The bio psycho-social model of disease may be helpful in describing the interaction between body, mind and circumstance. Putting the CFS patient at ease and explaining the pathophysiology of the symptoms is a useful approach but many patients and patient associations are still very somatically orientated, thereby sustaining the condition. However, in patients who accept that their problems may be stress-induced and are prepared to participate in therapy, some therapies have been proven to be effective, notably cognitive behavioural therapy. PMID- 15835624 TI - [Decisions around the end of life on Intensive Care: making the transition from curative to palliative treatment]. AB - The decision to move from curative treatment to palliative care in the intensive care situation is less related to morals and ethics than it is to the assessment of medical issues, professionalism, communication and orchestration. Treatment should be considered medically pointless if, in the view of the treating physicians, it does not offer realistic chance to return to a meaningful life. Continuing futile care can be seen as disrespectful, both to the patient, his partner and the family, as well as to the members of the ICU team. Intensivists are responsible for withholding or withdrawing life support to patients in whom further life support is considered futile and who are unable to express their wishes due to critical illness and sedation. The intensivist typically makes this type of decision after a period in which medical and other information has been collected and after intensive discussions with other medical professionals as well as the partner and family. This is based on the trust that is built up through their skill, attitude and behaviour and that is perpetuated in a continuing process of intensive communication. Conflicts should be prevented, or at least recognised early and discussed. Ifa conflict is ongoing then it should be tackled by planning a number of consecutive consultations. PMID- 15835625 TI - [Development in the treatment of malignant mesothelioma]. AB - For many years there has been a search for an effective treatment of malignant pleural mesothelioma. Surgery is of limited applicability and is reserved for special cases, in which it is combined with radiation therapy. In the previous century, many cytostatic agents have been tested, alone or in combination, but the response was limited, the median survival time was unchanged and the toxicity was high. New drugs, including the new antifolates, are being used much more often in the treatment of malignant pleural mesothelioma in combination with a platinum derivative. Cytostatics such as pemetrexed, an antifolate, and to a lesser extent raltitrexed, have shown good response rates and increased survival in phase III studies, but the survival benefit evaporates within 2 years. Inhibitors of angiogenesis and of epidermal growth factor are being tested, but they have shown only limited activity until now. The current studies focus on the use of chemotherapy and biological agents as part of a more complex treatment schedule. PMID- 15835626 TI - [Side effects of tramadol: 12 years of experience in the Netherlands]. AB - Tramadol is a synthetic opioid that has been available in the Netherlands since 1992 and is usually used as a centrally-acting analgesic when paracetamol or an NSAID provides insufficient relief. In the period 1 January 1992--30 November 2003, the Netherlands Pharmacovigilance Centre Lareb received 299 reports concerning 522 adverse drug reactions associated with the use of tramadol. Some of the frequently reported side effects with a high reporting odds ratio were nausea, constipation and withdrawal symptoms. These side effects are very similar to those of the other opioids due to the affinity of tramadol for the micro opioid receptor. Because tramadol is often not recognised as an opioid, it is important that such opiate effects be recognised as an adverse drug reaction on time. PMID- 15835627 TI - [Diagnostic image (234). A man with an acute scrotum]. AB - A 48-year-old man presented with acute testicular pain caused by a haematoma in a non seminoma testicular tumour. PMID- 15835628 TI - [Problematic cases of mesothelioma reported to the Dutch Institute for Asbestos Victims: evaluation by the Mesothelioma Working Group of the Netherlands Association of Pulmonologists and Specialists in Tuberculosis]. AB - OBJECTIVE: To analyse the assessments of problematic cases reported to the Dutch Institute for Asbestos Victims (IAS) by the Mesothelioma Working Party of the Netherlands Association of Pulmonologists and Specialists in Tuberculosis (NVALT). DESIGN: Descriptive. METHOD: The pathological confirmation of a malignant pleural mesothelioma of occupational origin is difficult in about 10% of the cases. The IAS has requested the Mesothelioma Working Party of the NVALT to review these cases. When no definitive diagnosis can be made on histological or cytological grounds, three pulmonologists reach a conclusion on the basis of correspondence, X-ray examination and other information. RESULTS: In the period January 2000--March 2004 the Working Party evaluated 132 cases, two-thirds of whom (n = 89) were assessed to be compatible with 'malignant pleural mesothelioma' and one-third of whom (n = 43) were felt to be non-compatible. In 69% of the cases (91/132) the conclusions of the three independent specialists were unanimous. The median time from request to report was 25 days (range: 1 185). CONCLUSION: This approach was effective and rapid. PMID- 15835629 TI - [Sexual harassment of medical students during their period of work placement]. AB - OBJECTIVE: To investigate the incidence, type and consequences of sexual harassment of medical students at the Radboud University in Nijmegen, the Netherlands, during their period of work placement, as well as the students' need for care thereafter. DESIGN: Questionnaire. METHOD: During the period from 1 July to 31 December 2003, 5th and 6th year medical students were asked about their experiences with sexual harassment by means of a questionnaire. Sexual harassment was defined as unwelcome, sexually-coloured attention. RESULTS: Of the 183 questionnaires distributed, 113 (62%) were returned. 15 (20%) of the 75 female students and none of the 38 male students had experienced sexual harassment. The offenders included 9 patients and 6 doctors (54 men and 1 woman). In 7 of the 15 cases the harassment consisted of combinations of unwanted behaviour and unwanted sexually-coloured remarks. 9 of the 15 students had discussed their experiences with their peers, 7 with a supervisor and 3 with nobody. The most important reason to discuss it first in their peer group was that, despite the fact that the students were convinced that the offender's behaviour was unacceptable, they still doubted their own judgement. 5 students felt inhibited in their contacts with patients after the incident. 8 of the 15 offenders were not confronted with their behaviour. 6 of the 15 students were not satisfied with the way their case was handled. CONCLUSION: The problem of sexual harassment of medical students during their period of work placement should not be underestimated. It has a negative impact on the personal and professional conduct of future doctors. This subject should be part of the training of both medical students and their supervisors. PMID- 15835630 TI - [Rocky Mountain spotted fever in an American tourist]. AB - In a 28-year-old male American tourist who presented in the hospital with fever, cold shivers, headache, nausea, myalgia and arthralgia, Rocky Mountain spotted fever was suspected, partly because he came from an endemic region (the state of Georgia). The patient was treated with doxycycline, 100 mg b.i.d.; 9 days after the first appearance of the symptoms, the diagnosis was confirmed by the report of a positive antibody titre against Rickettsia rickettsii. The patient did not have exanthema. He was discharged in good general condition after two weeks of treatment. Rocky Mountain spotted fever, caused by the Gram-negative bacterium R. rickettsii, is a serious rickettsiosis. The disease is seen only sporadically in the Netherlands because the ticks in the Netherlands do not carry the bacterium. The travel history is still not a standard component of the anamnesis and is therefore often forgotten. This can lead to under-diagnosis and delayed treatment of diseases that were formerly limited to the continent. The early recognition and treatment of Rocky Mountain spotted fever is important since delayed treatment is associated with a clear increase in both morbidity and mortality. PMID- 15835631 TI - [Pseudogout in 3 patients with presumed therapy-resistant rheumatoid arthritis]. AB - 3 patients, 2 women aged 71 and 76 and a 55-year-old man, were originally diagnosed with rheumatoid arthritis (RA) and treated with disease-modifying antirheumatic drugs (DMARDs); two of these patients fulfilled the American College of Rheumatology criteria for RA. Because the symptoms persisted, the diagnosis was reconsidered. It turned out that they had pseudogout, which is an arthropathy caused by the deposition of calcium pyrophosphate crystals; the younger woman had no obvious metabolic disorder, the older woman had underlying hyperparathyroidism, and in the man the arthropathy was probably due to benign hypercalcaemia. DMARDs were replaced by NSAIDs. Varying degrees ofarthropathy persisted in the women, but in the man they were clearly decreased. In addition to resembling gout, the clinical manifestations of pseudogout can also mimic RA. It is important to distinguish pseudogout from RA because their treatment is completely different. Furthermore, pseudogout can be the first or sole symptom of a metabolic disorder. PMID- 15835632 TI - [Results from 10 years of preimplantation-genetic diagnosis in The Netherlands]. PMID- 15835633 TI - [Nonalcoholic steatohepatitis: diagnosis, pathogenesis, treatment and prognosis]. PMID- 15835634 TI - [Congenital heart disease and pregnancy]. PMID- 15835635 TI - [Cost of foods fortified with plant sterols refunded by a health insurance company]. PMID- 15835636 TI - [Cost of foods fortified with plant sterols refunded by a health insurance company]. PMID- 15835637 TI - [Cost of foods fortified with plant sterols refunded by a health insurance company]. PMID- 15835639 TI - Choosing your retirement journey. PMID- 15835640 TI - Retirement: the beginning or the end. PMID- 15835641 TI - Nearing retirement? Give yourself a financial check-up. PMID- 15835642 TI - 10 rules for the retirement road. PMID- 15835643 TI - Retirement prescriptions. PMID- 15835644 TI - The emotional side of retirement planning. PMID- 15835646 TI - Preparation of oleic acid/water emulsions in surfactant-free condition by sequential processing using midsonic-megasonic waves. PMID- 15835647 TI - Glued Langmuir-Blodgett film: an unexpected dependency of gluing on polyelectrolyte concentration. PMID- 15835648 TI - Lithography with a focused soft X-ray beam and a monomolecular resist. PMID- 15835649 TI - A new interpretation of serum albumin surface passivation. PMID- 15835650 TI - Polyelectrolyte-quantum dot multilayer films fabricated by combined layer-by layer assembly and Langmuir-Schaefer deposition. PMID- 15835651 TI - Unsaturation effect on gelation behavior of aryl glycolipids. AB - Structurally simple, renewable-resource-based cardanyl (glucoside)s [1, 1-O-3'-n (pentadecyl) phenyl-beta-D-glucopyranoside; 2, 1-O-3'-n-(8'(Z)-pentadeceenyl) beta-glucopyranoside; 3, 1-O-3'-n-(8'(Z),11'(Z)pentadecadienyl)phenyl-beta-D glucopyranoside, 4; 1-O-3'-n-(8'(Z),11'(Z),14'-pentadecatrienyl)phenyl-beta-D glucopyranoside; and the mixture 5] form thermally reversible transparent gels in a water/alcohol mixture and a number of organic solvents, strongly influenced by the unsaturation of the aliphatic alkyl chain. DSC studies revealed that the Tgel of 1 in water/ethanol (1:1, vol/vol) gel is 69.0 degrees C, while of the introduction of a single double bond reduces the value to 30.0 degrees C in the case of monoene 2, indicating that the stability of the gel is related to the number of double bonds on the lipophilic part of the gelator. Furthermore, XRD measurements showed that the aqueous gel 1 maintains an interdigitated bilayered structure with 3.14 nm long-range ordering, and the corresponding organogel maintains an extended bilayer structure for 4.34 nm, indicating a clear difference in the aggregation behavior in different solvents. PMID- 15835652 TI - Colloidal stability of aqueous dispersions of block ionomer complexes: effects of temperature and salt. AB - This work characterized colloidal stability of the dispersions, formed by the complexes of poly(ethylene oxide)-b-poly(sodium methacrylate) and hexadecyltrimethylammonium bromide. At room temperature, the dispersion was stabilized by the poly(ethylene oxide) (PEO) chains and did not aggregate for at least several months. Elevation of temperature caused aggregation of the dispersion because of dehydration of the PEO chains. At initial stages (minutes), the aggregation was reversible and the particles spontaneously redispersed once the temperature was decreased. However, it became irreversible at the later stages (hours), probably indicating fusion of the hydrophobic cores of the BIC particles. Addition of elementary salts led to a decrease of the aggregation temperature. The effects of various salts were dependent on the chemical nature of the ions and were consistent with the Hofmeister series. This behavior was discussed in terms of hydration and London (dispersion) interactions between the ions and the PEO. PMID- 15835653 TI - Evaporation rates of water from concentrated oil-in-water emulsions. AB - We have investigated the rate of water evaporation from concentrated oil-in-water (o/w) emulsions containing an involatile oil. Evaporation of the water continuous phase causes compression of the emulsion with progressive distortion of the oil drops and thinning of the water films separating them. Theoretically, the vapor pressure of water is sensitive to the interdroplet interactions, which are a function of the film thickness. Three main possible situations are considered. First, under conditions when the evaporation rate is controlled by mass transfer across the stagnant vapor phase, model calculations show that evaporation can, in principle, be slowed by repulsive interdroplet interactions. However, significant retardation requires very strong repulsive forces acting over large separations for typical emulsion drop sizes. Second, water evaporation may be limited by diffusion in the network of water films within the emulsion. In this situation, water loss by evaporation from the emulsion surface leads to a gradient in the water concentration (and in the water film thickness). Third, compression of the drops may lead to coalescence of the emulsion drops and the formation of a macroscopic oil film at the emulsion surface, which serves to prevent further water evaporation. Water mass-loss curves have been measured for silicone o/w emulsions stabilized by the anionic surfactant SDS as a function of the water content, the thickness of the stagnant vapor-phase layer, and the concentration of electrolyte in the aqueous phase, and the results are discussed in terms of the three possible scenarios just described. In systems with added salt, water evaporation virtually ceases before all the water present is lost, probably as a result of oil-drop coalescence resulting in the formation of a water-impermeable oil film at the emulsion surface. PMID- 15835654 TI - A small-angle neutron scattering study of cholic acid-based organogel systems. AB - Small-angle neutron scattering measurements were performed on some cholic acid based gel systems in order to gain detailed information about the network structure. The presence of thin fibers with a radius of about 10-20 A was found for various gelators. Two types of interaction between different sorts of fibers were demonstrated, depending on the molecular structure of the gelator. The first type involves the presence of microcrystalline knots with a dimension of about 100-200 A between the fibers. Upon heating, this network gradually disintegrates. The second type involves loose entanglements between flattened fibers. The occurrence of these types of interaction is related to the length of the alkyl tail attached to cholic acid. PMID- 15835655 TI - Flow cytometry: a new method to investigate the properties of water-in-oil-in water emulsions. AB - We present a new and facile method to evaluate w/o/w emulsions containing fluorescent markers by flow cytometry. Flow cytometry allows simultaneous measurement of w/o/w emulsion droplets "marked" with a fluorescent marker or "blank" without the need for complicated sample preparation. The yield of preparation of the w/o/w emulsion and the release rate of the fluorescent marker FITC-BSA were investigated by this new method. The release fraction (after 24 h) of FITC-BSA from the w/o/w emulsion decreased with increasing concentration of FITC-BSA inside the internal phase, just like the release fraction of NaCl as marker from the w/o/w emulsion. Flow cytometry results show that the yield and release behavior in w/o/w emulsions are in agreement with results reported by more complicated methods. PMID- 15835656 TI - Thermal gelation of cellulose in a NaOH/thiourea aqueous solution. AB - Utilizing a novel solvent of cellulose, 6 wt % NaOH/5 wt % thiourea aqueous solution, for the first time, we prepared the thermally induced cellulose gel. We investigated the thermal gelation of cellulose solutions with rheometry and the structure of the gel with 13C NMR, wide-angle X-ray diffraction, environmental scanning electron microscopy, and atomic force microscopy. The cellulose solutions revealed an increase in both the storage modulus (G') and the loss modulus (G") with an increase in the temperature during gelation. The temperature at the turning point, where G' overrides G" because of the onset of gelation, decreased from 38.6 to 20.1 degrees C with an increase of cellulose concentration from 4 to 6 wt %. Given enough time, G' of all solutions can exceed G" at a certain temperature slightly lower than the gelation temperature, indicating that the occurrence of the gelation is also a function of time. Each of the assigned peaks of NMR of the cellulose gel is similar to that of the cellulose solution, suggesting that the gelation resulted from a physical cross-linking. The gels were composed of relatively stable network units with an average diameter of about 47 nm. At either a higher temperature (at 60 degrees C for 30 s) or a longer gelation time (at 30 degrees C for 157 s), the gel in the 5 wt % cellulose solution could form. A schematic gelation process was proposed to illustrate the sol-gel transition: the random self-association of the cellulose chains having the exposed hydroxyl in the aqueous solution promotes the physical cross-linking networks. PMID- 15835657 TI - Synthesis and light scattering study of microgels with interpenetrating polymer networks. AB - Monodispersed microgels composed of poly(acrylic acid) (PAAc) and poly(N isopropylacrylamide) (PNIPAM) interpenetrating polymer networks (IPN) were synthesized by a two-step method, first preparing PNIPAM microgel and then polymerizing acrylic acid that interpenetrates into the PNIPAM network. The growth kinetics of the IPN particle formation was obtained by measuring the turbidity and particle hydrodynamic radius (Rh) as a function of reaction time. IPN and PNIPAM microgels were characterized and compared by dynamic and static light scattering techniques. The concentrated aqueous solutions of the PNIPAM PAAc IPN microgels exhibit an inverse thermoreversible gelation. In contrast to polymer solutions of poly(NIPAM-co-AAc) that have the inverse thermoreversible gelation, our system can self-assemble into an ordered structure, displaying bright colors. Furthermore, IPN microgels undergo the reversible volume phase transitions in response to both pH and temperature changes associated with PAAc and PNIPAM networks, respectively. PMID- 15835658 TI - Controlled, rapid deposition of structured coatings from micro- and nanoparticle suspensions. AB - The objective of the study was to develop the operational basis for rapid and controlled deposition of crystal coatings from particles of a wide size range. We deposited such structured coatings by dragging with constant velocity a small volume of liquid confined in a meniscus between two plates. Two types of structured coatings were characterized: latex colloidal crystals and thin layers from metallic nanoparticles. The crystal deposition was sped up by use of preconcentrated suspensions. Crystal coatings larger than a few square centimeters were deposited in minutes from aqueous suspension volumes of approximately 10 microL. The governing mechanism of crystal deposition is convective assembly at high volume fractions. The two major process parameters that allow control over the coating thickness and structure were the deposition speed and particle volume fraction. The evaporation rate was not found to affect the process to a large extent. A volumetric flux balance was used to relate the deposition parameters to coating structure and properties. Operational "phase" diagrams were constructed, relating the crystal layer thickness and packing symmetry to the process parameters. These diagrams could be instrumental in transforming the convective colloidal deposition into a robust scaleable technology. PMID- 15835659 TI - Two-dimensional crystallization of microspheres by a coplanar AC electric field. AB - The particle-field and particle-particle interactions induced by alternating electric fields can be conveniently used for on-chip assembly of colloidal crystals. Two coplanar electrodes with a millimeter-sized gap between them are used here to assemble two-dimensional crystals from suspensions of either latex or silica microspheres. When an AC voltage is applied, the particles accumulate and crystallize on the surface between the electrodes. Light diffraction and microscopic observations demonstrate that the hexagonal crystal is always oriented with one axis along the direction of the field. The particles disassemble when the field is turned off, and the process can be repeated many times. The diffraction patterns from all consecutively formed crystals are identical. This assembly is driven by forces that depend on the electric field gradient, and a model is proposed involving a combination of dielectrophoresis and induced dipole chaining. The organization of large two-dimensional crystals allows characterization of the electrostatic interactions in the particle ensembles. The process can be controlled via the field strength, the frequency, and the viscosity of the liquid media. It could be used to make rudimentary optical switches or to separate mixtures of particles of different sizes. PMID- 15835660 TI - Temperature-dependent vesicle formation of aqueous solutions of mixed cationic and anionic surfactants. AB - The phase behavior of aqueous solutions of mixed cetyltrimethylammonium bromide (CTAB) and sodium octyl sulfate (SOS) was examined at different temperatures (20, 30, 40, and 50 degrees C). While stable vesicles were formed in a narrow composition range on the SOS-rich side at 20 degrees C, the range widened remarkably when the temperature was raised to 30 degrees C. Thus, the vesicle region extended to cover almost the entire composition range, CTAB:SOS = 0.5:9.5 5.0:5.0, at the total surfactant concentrations of 50-70 mM on the SOS-rich side. To analyze the temperature dependence of this phase behavior of the mixed surfactant system, DSC and fluorescence polarization measurements were performed on the system. The experimental findings obtained revealed that pseudo-double tailed CTAB/SOS complex, the major component of the bimolecular membrane formed by the surfactant mixture, undergoes a gel (Lbeta)-liquid crystal (Lalpha) phase transition at about 26 degrees C. This phenomenon was interpreted as showing that the bimolecular membrane has no curvature and is rigid and easy to precipitate at temperatures below the phase transition point, whereas it has a curvature and is loose enough to disperse in the solution as vesicles at temperatures above the phase transition point. Vesicles formed by the anionic/cationic surfactant complex were then stable at temperatures above the phase transition temperature of the complex. PMID- 15835661 TI - Functional group distributions in carboxylic acid containing poly(N isopropylacrylamide) microgels. AB - Control of the functional group distribution is of fundamental importance in the design of functional polymer particles, particularly in biological applications. Surface-functionalized particles are useful for bioconjugation and medical diagnostics, while internally functionalized particles may have applications in drug delivery. We have prepared a series oftemperature-sensitive poly(N isopropylacrylamide) (PNIPAM)-based microgels containing carboxylic acid functional groups via copolymerization with methacrylic acid and acrylamide, which was selectively hydrolyzed under optimized conditions to generate the carboxylic acid functionality. The resulting microgels were analyzed using conductometric and potentiometric titration, dynamic light scattering, and electrophoresis. Acrylamide-containing microgels hydrolyzed below the volume phase transition temperature (VPTT) show broad particle size versus temperature profiles, relatively low electrophoretic mobilities at basic pH, and time dependent base titration profiles, suggesting the presence of internal functional groups whose titration is diffusion-controlled. Methacrylic acid containing microgels show sharper particle size versus temperature profiles, higher electrophoretic mobilities at basic pH, and time-independent base titration profiles, suggesting the presence of a "core-shell" structure with primarily surface functionalization. Similar results were obtained when acrylamide containing microgels were hydrolyzed at temperatures above the VPTT. Thus, through selection of comonomer and hydrolysis conditions, we have developed strategies to control and characterize the number and distribution ofcarboxylic acid functional groups in PNIPAM-based microgels. PMID- 15835662 TI - Simultaneous conductivity and viscosity measurements as a technique to track emulsion inversion by the phase-inversion-temperature method. AB - Two kinds of transitions can occur when an emulsified water-oil-ethoxylated nonionic surfactant system is cooled under constant stirring. At a water-oil ratio close to unity, a transitional inversion takes place from a water-in-oil (W/O) to an oil-in-water (O/W) morphology according to the so-called phase inversion-temperature method. At a high water content, a multiple w/O/W emulsion changes to a simple O/W emulsion. The continuous monitoring of both the emulsion conductivity and viscosity allows the identification of several phenomena that take place during the temperature decrease. In all cases, a viscosity maximum is found on each side of the three-phase behavior temperature interval and correlates with the attainment of extremely fine emulsions, where the best compromise between a low-tension and a not-too-unstable emulsion is reached. The studied system contains Polysorbate 85, a light alkane cut oil, and a sodium chloride brine. All transitions are interpreted in the framework of the formulation-composition bidimensional map. PMID- 15835663 TI - Shape fluctuations of large unilamellar lipid vesicles observed by laser light scattering: influence of the small-scale structure. AB - In the present paper, we apply the dynamic laser light scattering technique to investigate the dependence of the characteristic times of thermally induced shape fluctuation of large unilamellar vesicles (LUVs) on bilayer composition. After addressing single-component LUVs made of two common phospholipids, dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), we investigate the changes in vesicle shape fluctuation times due to the presence of cholesterol and gangliosides (GM1), added in small amounts. The experimental results show that the addition of a second component, even in small amount, to DMPC vesicles induces a change in membrane fluctuation times. Moreover, in the case of ganglioside addition, also the disposition of GM1 within the bilayer is of importance. Quite unexpectedly, the symmetric or asymmetric disposition of GM1 has opposite effects on bilayer dynamics, the first resulting in a "hardening" and the second in a "softening" of the membrane. Those results support that the small-scale structure of the bilayer is important in determining the overall dynamics of the vesicle. They also suggest that the physiological disposition of GM1 in the outer leaflet of real cells has a significative result in mechanical terms, positively affecting the dynamics of the membrane. PMID- 15835664 TI - Electrophoresis of a concentrated dispersion of spherical particles in a non Newtonian fluid. AB - Electrophoresis is one of the most widely used analytical tools for the quantification of the charged conditions on the surface of fine particles including biological entities. Although it has been studied extensively in the past, relevant results for the case when the dispersion medium is non-Newtonian are very limited. This may occur, for example, when the concentration of the dispersed phase is not low, which is not uncommon in practice. Here, the electrophoresis of a concentrated spherical dispersion in a Carreau fluid is analyzed theoretically under the conditions of low electric potential and weak external applied electrical field. A pseudospectral method coupled with a Newton Raphson iteration procedure is used to solve the electrokinetic equations describing the phenomenon under consideration. We conclude that the more significant the shear thinning effect of the fluid, the larger the mobility, and this phenomenon is pronounced for the case when the double layer surrounding a particle is thin. We show that if the double layer is thin and the effect of shear thinning is significant, a second vortex can be observed in the neighborhood of a particle. PMID- 15835665 TI - Association behavior of poly(methyl methacrylate-block-methacrylic acid) in aqueous medium. AB - The atom transfer radical polymerization technique was used to synthesize the poly(methyl methacrylate-block-methacrylic acid) (P(MAA-b-MMA)) copolymer in order to study the aggregation behavior in aqueous solution over the course of neutralization. Combinations of static and dynamic light scattering (SLS, DLS) and potentiometric titration techniques were used to investigate the size and shape of the micelle at various degrees of neutralization (alpha). By comparing the effect of different polymer chain length with similar MMA/MAA ratio on the aggregation behavior during neutralization, we found relatively strong entanglement of long MMA polymer chains. The comparison between the different MMA/MAA ratios showed that longer MMA chains produced more entanglements. Conductometric titration was used to determine the counterion condensation phenomenon during the course of neutralization. At a critical micellar charge density observed at alpha approximately 0.4, Na+ ions are condensed on the polymer chains. The amount of condensed Na+ was evaluated by the conductivity change, yielding the condensation ratio when the polymer was completely neutralized. PMID- 15835666 TI - Phase behavior of a mixture of poly(isoprene)-poly(oxyethylene) diblock copolymer and poly(oxyethylene) surfactant in water. AB - The phase behavior of a mixture of poly(isoprene)-poly(oxyethylene) diblock copolymer (PI-PEO or C250EO70) and poly(oxyethylene) surfactant (C12EO3, C12EO5, C12EO6, C12EO7, and C12EO9) in water was investigated by phase study, small-angle X-ray scattering, and dynamic light scattering (DLS). The copolymer is not soluble in surfactant micellar cubic (I1), hexagonal (H1), and lamellar (Lalpha) liquid crystals, whereas an isotropic copolymer fluid phase coexists with these liquid crystals. Although the PI-PEO is relatively lipophilic, it increases the cloud temperatures of C12EO3-9 aqueous solutions at a relatively high PI-PEO content in the mixture. Most probably, in the copolymer-rich region, PI-PEO and C12EOn form a spherical composite micelle in which surfactant molecules are located at the interface and the PI chains form an oil pool inside. In the C12EO5/ and C12EO6/PI-PEO systems, one kind of micelles is produced in the wide range of mixing fraction, although macroscopic phase separation was observed within a few days after the sample preparation. On the other hand, small surfactant micelles coexist with copolymer giant micelles in C12EO7/ and C12EO9/PI-PEO aqueous solutions in the surfactant-rich region. The micellar shape and size are calculated using simple geometrical relations and compared with DLS data. Consequently, a large PI-PEO molecule is not soluble in surfactant bilayers (Lalpha phase), infinitely long rod micelles (H1 phase), and spherical micelles (I1 phase or hydrophilic spherical micelles) as a result of the packing constraint of the large PI chain. However, the copolymer is soluble in surfactant rod micelles (C12EO5 and C12EO6) because a rod-sphere transition of the surfactant micelles takes place and the long PI chains are incorporated inside the large spherical micelles. PMID- 15835667 TI - Surfactant diffusion through bicontinuous micellar networks: a case study of the C9G1/C10G1/H2O mixed surfactant system. AB - Self-diffusion coefficients were obtained by means of NMR diffusometry for differing ratios of n-decyl-beta-D-glucopyranoside (C10G1) and n-nonyl-beta-D glucopyranoside (C9G1) surfactant mixtures, along dilution lines through the micellar region of the ternary C9G1/C10G1/H2O phase diagram. Networks of bicontinuous micelles have been suggested to exist throughout the micellar regions of the phase diagram. A phase separation into two coexisting liquid solutions is observed in the dilute, C10G1-rich regions of the phase diagram. The fact that the dilution curves follow scaling relations pertaining to surfactant diffusion in a network for mixtures rich in C10G1 indicates that the phase separation is due to differences in the networks in different micellar regions of the phase diagram; networks remain largely intact despite dilution down to the phase separation in the C10G1-rich region, whereas networks with scissions are predicted to exist in the C9G1-rich regions of the micellar phase. PMID- 15835668 TI - Isothermal titration calorimetric studies on the temperature dependence of binding interactions between poly(propylene glycol)s and sodium dodecyl sulfate. AB - Isothermal titration calorimetry (ITC) is a sensitive research tool for examining the binding interactions between surfactant and polymer where the differential enthalpy during the binding process is monitored. In addition to the critical micelle concentration (cmc) and the micellization enthalpy (deltaHm), the effective micellar charge fraction (beta) of the ionic surfactant micellization process can also be determined from ITC thermograms. Poly(propylene glycol) (PPG) exhibits a lower critical solution temperature (LCST) ranging from 15 to 42 degrees C, depending on the molecular weights. We report, for the first time, the binding interactions between sodium dodecyl sulfate (SDS) and 1,000, 2,000 and 3,000 Da PPGs, where different binding mechanisms are in operation, depending on the temperature. At temperatures lower than the LCST, the binding interactions are similar to those of SDS and low molecular weight poly(ethylene glycol)s (MW < 3500 Da). At temperatures greater than the LCST, the binding interactions are dominated by direct solubilization of PPG chains into mixed micellar cores. At temperatures near the LCST, the binding interactions are controlled by the balance ofthe PPG solubilization at low SDS concentrations and polymer-induced micellization at high SDS concentrations. PMID- 15835669 TI - Synthesis of vinyl polymer-silica colloidal nanocomposites prepared using commercial alcoholic silica sols. AB - The surfactant-free synthesis of vinyl polymer-silica nanocomposite particles has been achieved in aqueous alcoholic media at ambient temperature in the absence of auxiliary comonomers. Styrene, methyl methacrylate, methyl acrylate, n-butyl acrylate, and 2-hydroxypropyl methacrylate were homopolymerized in turn in the presence of three commercially available ultrafine alcoholic silica sols. Stable colloidal dispersions with reasonably narrow size distributions were obtained, with silica contents of up to 58% by mass indicated by thermogravimetric analysis. Particle size distributions were assessed using both dynamic light scattering and disk centrifuge photosedimentometry. The former technique indicated that the particle size increased for the first 1-2 h at 25 degrees C and thereafter remained constant. Particle morphologies were studied using electron microscopy. Most of the colloidal nanocomposites comprised approximately spherical particles with relatively narrow size distributions, but in some cases more polydisperse or nonspherical particles were obtained. Selected acrylate based nanocomposites were examined in terms of their film formation behavior. Scanning electron microscopy studies indicated relatively smooth films were obtained on drying at 20 degrees C, with complete loss of the original particle morphology. The optical clarity of solution-cast 10 microm nanocomposite films was assessed using visible absorption spectrophotometry, with 93-98% transmission being obtained from 400 to 800 nm; the effect of long-term immersion of such films in aqueous solutions was also examined. X-ray photoelectron spectroscopy studies indicated that the surface compositions of these nanocomposite particles are invariably silica-rich, which is consistent with their long-term colloidal stability and also with aqueous electrophoresis measurements. FT-IR studies suggested that in the case of the poly(methyl methacrylate)-silica nanocomposite particles, the carbonyl ester groups in the polymer are hydrogen-bonded to the surface silanol groups. According to differential scanning calorimetry studies, the glass transition temperatures of several poly(methyl methacrylate)-silica and polystyrene-silica nanocomposites can be either higher or lower than those of the corresponding homopolymers, depending on the nature of the silica sol. PMID- 15835670 TI - Aggregation behavior of aqueous solutions of ionic liquids. AB - The aggregation behavior in aqueous solutions of three ionic liquids based on the 1-alkyl-3-methylimidazolium cation has been investigated by means of surface tension, conductivity, and small-angle neutron scattering (SANS) measurements. From analysis of the SANS data, models for the shapes and sizes of aggregates have been proposed: the short-chain 1-butyl-3-methylimidazolium tetrafluoroborate [C4mim] [BF4] system can be best modeled by treating it as a dispersion of polydisperse spherical aggregates that form above a critical aggregation concentration, whereas the 1-octyl-3-methylimidazolium iodide, [C8mim] [I], solutions can be modeled as a system of regularly sized near-spherical charged micelles that form above a critical micelle concentration. Solutions of 1-octyl-3 methylimidazolium chloride, [C8mim]-[Cl], display weak long-range ordering of possibly disklike particles culminating in the formation of structures with distinct long-range order at higher concentrations. PMID- 15835671 TI - Temperature dependence of the thermodynamics and kinetics of micellar solutions. AB - We predict theoretically the thermodynamics and relaxation kinetics of solutions of cylindrical branched micelles. Using a recently developed theory in combination with the experimental data, we explain the unusual, inverted temperature dependence of the phase separation observed in wormlike micelles and dilute microemulsions. We extend the model to treat the temperature dependence of the relaxation kinetics and explain the observations. PMID- 15835672 TI - Gels of hydrophobically modified ethyl(hydroxyethyl) cellulose cross-linked by amylose: effects of hydrophobe architecture. AB - Previous work has shown that amylose (AM) can cross-link hydrophobically modified polymers by inclusion complexation, whereby thermoreversible cold-setting gels are formed. Here we investigate the complexation of AM with different samples of hydrophobically modified ethyl(hydroxyethyl) cellulose (HMEHEC), distinguished by differences in the architecture of the hydrophobes (the hydrophobic side chains). All hydrophobes, except one, were based on linear alkyl chains, but with varying chain lengths (C12-C14). In addition, some samples contained short hydrophilic "spacers", consisting of 2-5 ethylene oxide units, between the alkyl chains and the EHEC backbone. Gels of varying strength were obtained for the different AM/HMEHEC samples. The alkyl chain length seemed to be the major factor affecting the gel strength, with longer alkyl chains giving stronger gels. For similar alkyl chain lengths, stronger gels were obtained when a spacer was present. Addition of AM caused a small increase of the cloud points of HMEHECs with C14 hydrophobes in water. Time-dependent effects and effects of the sample preparation procedure were also investigated. The reversibility of the gelation with respect to shear was confirmed. A gel destroyed by added surfactant was shown to reform on removal of the surfactant by dialysis. PMID- 15835673 TI - Time-resolved synchrotron X-ray scattering of the crystallization of a soft hexagonal columnar crystal. AB - We have recently shown (Langmuir 2000, 16, 5846) that a soft surfactant hexagonal phase exhibits, above a critical shear rate, a shear-melting transition, where a two-dimensional polycrystalline texture is converted into a liquid of rods aligned along the flow. Moreover, after abrupt cessation of high shear, a monocrystal-like structure is obtained. In this paper, we investigate the kinetics of crystallization from the shear-melted phase by means of time-resolved synchrotron X-ray scattering experiments. After a latency time, the sample is found to continuously evolve from a liquid of rods to a monocrystal of rods. We show that the crystallization results from a coupling between bulk crystallization and interfacial crystallization, due to a specific anchoring of the liquid crystal at the walls of the shear cell. A detailed analysis of the experimental data allows the evaluation of the propagation of the interfacial front and the time evolution of the size of the crystallites. We find that the size scales as t(v) with v approximately equal to 0.3, in fair agreement with recent theories and simulations on grain rotation-induced grain growth of columnar polycrystalline structures. PMID- 15835674 TI - Study of Ca-ATMP precipitation in the presence of magnesium ion. AB - ATMP (aminotri(methylenephosphonic acid)), a phosphonate scale inhibitor used in the petroleum industry, was used as a model scale inhibitor in this study. One of the goals of this work was to determine the range of conditions under which Mg ions, which are formed in reservoir formations containing dolomite, modulate the formation of Ca-ATMP precipitate as a scale inhibitor. The results revealed that the amount of ATMP precipitated decreased with addition of Mg ions in solution at all values of the solution pH. Furthermore, an increase in both the solution pH and the concentration of the divalent cations in solution resulted in a change of the molar ratio of (Ca + Mg) to ATMP in the precipitates. At a low solution pH (pH 1.5), Mg ions had little effect on the composition of the Ca-ATMP precipitate. However, at higher values of the solution pH (pH 4 and 7), the Ca to ATMP molar ratio in the precipitates decreased with increasing concentration of the Mg. Here it was found that Mg ions replaced Ca ions on available reactive sites of ATMP molecules. These results determined the limits of the Mg ion concentration, which affects the precipitation of Ca-ATMP, Mg-ATMP, and (Ca + Mg) ATMP. The dissolution of the scale inhibitors was studied using a rotating disk reactor. These experiments showed that the total divalent cation molar ratio (Ca + Mg) to ATMP in the precipitates is the primary factor that controls the rate of dissolution (release) of the phosphonate precipitates. The phosphonate precipitate dissolution rates decreased as the molar ratio of divalent cations to ATMP in the precipitates increased. PMID- 15835675 TI - Adhesion force of a wedge. AB - The macroscopic theory of capillarity is usually applied to evaluate the adhesion forces produced by fluid bridges between solid particles. A refined mesoscopic description incorporates the disjoining pressure, which results from the long range forces between the fluid and the solid substrates. In the case of simple nonpolar fluids the influence of the disjoining pressure on adhesion is expected to be negligibly small. Water bridges, however, sometimes have very large disjoining pressures on various substrates. Then mesoscopic theory leads to pull off forces which differ from the predictions of the macroscopic approach. PMID- 15835676 TI - Acid/Base-treated activated carbons: characterization of functional groups and metal adsorptive properties. AB - Surface modification of activated carbons by various physicochemical methods directs an attractive approach for improvement of heavy metal uptake from aqueous solutions. Activated carbons were modified with HCl and HNO3 optionally followed by NaOH. The effects of surface modifications on the properties of the carbons were studied by the specific surface area, carbon pH, and total acidity capacity as well as by scanning electron microscopy, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy. The modifications bring about substantial variation in the chemical properties whereas the physical properties remain nearly unchanged. NaOH causes an increase in the content of hydroxyl groups, while the HCl treatment results in an increase in the amount of single bonded oxygen functional groups such as phenols, ethers, and lactones. The HNO3 modification generates a large number of surface functional groups such as carbonyl, carboxyl, and nitrate groups. The HNO3 modification significantly increases the copper adsorption, while the HCl treatment slightly reduces the copper uptake. Most of the copper ions are adsorbed rapidly in the first 2 h; the adsorption equilibrium is established in around 8 h. An intraparticle diffusion model successfully describes the kinetics of copper adsorption onto the carbons. PMID- 15835677 TI - Mineralization mechanism of calcium phosphates under three kinds of Langmuir monolayers. AB - Three kinds of Langmuir monolayers formed by dipalmitoylphosphatidylcholine (DPPC), arachidic acid (AA), and octadecylamine (ODA) were used as templates to study the initial stage of nucleation and crystallization of calcium phosphates. It was demonstrated that the combination of calcium ions (or phosphates) to the monolayer/subphase interface is a prerequisite for subsequent nucleation. It was found that calcium phosphate dihydrate (DPCD) formed at 25.0 degrees C for 12 h has a biphasic structure containing both amorphous and crystalline phases. These results showed that calcium phosphates were formed through a multistage assembly process, during which an initial amorphous phase DPCD was followed by a phase transformation into a crystalline phase and then the most stable hydroxyapatite (HAp). This provided new insights into the template-biomineral interaction and a mechanism for biomineralization. PMID- 15835678 TI - Chemical imaging of terrace-based active sites on gold. AB - Scanning tunneling microscopy data of a mixed monolayer comprised of a 40:60 ratio of H8Si8O12 and C6H13-H7Si8O12 clusters on gold are presented. The images display a composite monolayer surface with well-defined domain regions of the individual components. Holes present at face-centered cubic (fcc) sites of the starting Au/H7Si8O12 adsorbate layer indicate the location of active sites for impinging C6H13-H7Si8O12 clusters. Adsorption of a C6H13-H7Si8O12 cluster likely yields a mobile hydrogen atom available to recombine with and desorb an adjacent H8Si8O12 cluster. Hydrogen atom diffusion along substrate [121] directions is the proposed pattern formation mechanism of the mixed monolayer. Imaging of the spherosiloxane cluster domains identifies a novel terrace-based active site located in the fcc regions of the Au(111) 23 x square root3 surface reconstruction. PMID- 15835679 TI - Selective atomic layer deposition of titanium oxide on patterned self-assembled monolayers formed by microcontact printing. AB - We demonstrate a selective atomic layer deposition of TiO2 thin films on patterned alkylsiloxane self-assembled monolayers. Microcontact printing was done to prepare patterned monolayers of the alkylsiloxane on Si substrates. The patterned monolayers define and direct the selective deposition of the TiO2 thin film using atomic layer deposition. The selective atomic layer deposition is based on the fact that the TiO2 thin film is selectively deposited only on the regions exposing the silanol groups of the Si substrates because the regions covered with the alkylsiloxane monolayers do not have any functional group to react with precursors. PMID- 15835680 TI - Lateral and vertical nanophase separation in Langmuir-Blodgett films of phospholipids and semifluorinated alkanes. AB - It has recently been found that monodisperse surface micelles (hemimicelles) were formed in Langmuir monolayers of the semifluorinated alkane C8F17C16H33 (F8H16) after transfer onto silicon wafers. Grazing incidence X-ray diffraction studies have demonstrated that compression of mixed Langmuir monolayers made from combinations of dipalmitoyl phosphatidylethanolamine (DPPE) and diblock F8H16 in various molar ratios resulted in the complete expulsion of the diblock molecule at high surface pressure. F8H16 then formed a second layer on top of a DPPE-only monolayer, demonstrating a novel type of reversible, pressure-induced, vertical phase separation. Using atomic force microscopy and X-ray reflectivity, we show now that mixed DPPE/F8H16 (1:1.3) Langmuir-Blodgett films transferred onto silicon wafers below 10 mN m(-1) are laterally phase separated and consist of domains of F8H16 surface micelles in coexistence with a monolayer of DPPE. The density of the network of F8H16 surface micelles increases when the surface pressure of transfer increases. Around 10 mN m(-1), the F8H16 surface micelles start to glide on the DPPE monolayer, progressively overlying it, until total coverage is achieved. PMID- 15835681 TI - Surface ordering in dilute dihexadecyl dimethyl ammonium bromide solutions at the air-water interface. AB - At elevated temperatures and in dilute solution, we have observed lamellar surface ordering at the air-water interface of dihexadecyl dimethylammonium bromide, DHDAB, in the presence of electrolyte. With increasing temperature, the onset in ordering is observed between 35 and 40 degrees C. At 40 degrees C, there is an abrupt change in the lamellar spacing, from approximately 33 to approximately 40 A. Furthermore, in the presence of the cosurfactant benzyl alcohol, the ordering occurs at a lower temperature, between 20 and 25 degrees C. The change in lamellar spacing with temperature is attributed to a surface induced transition, similar to the Lbeta to Lalpha phase transition observed in bulk lamellar dispersions. PMID- 15835682 TI - Hydrolytic stability of organic monolayers supported on TiO2 and ZrO2. AB - The hydrolytic stability of C18 monolayers supported on TiO2 and ZrO2 was studied. Three types of monolayers were prepared from the following octadecyl modifiers: (1) octadecyldimethylchlorosilane (C18H37Si(CH3)2Cl); (2) octadecylsilane (C18H37SiH3); and (3) octadecylphosphonic acid (C18H37P(O)(OH)2). The hydrolysis of the surfaces prepared was studied under static conditions at 25 and 65 degrees C at pH 1-10. On the basis of the loss of grafted material, the stability of the monolayers fall in the following range: C18H37P(O)(OH)2 > or = C18H37SiH3 >> C18H37Si(CH3)2Cl. At 25 degrees C, monolayers from C18H37P(O)(OH)2 showed only approximately 2-5% loss in grafting density after one week at pH 1 10. The high stability of these monolayers was explained because of the strong interactions of the phosphonic acids with the substrates. Monolayers from C18H37Si(CH3)2Cl showed poor hydrolytic stability at any pH, which was explained because of the low stability of Ti-O-Si and Zr-O-Si bonds. Unlike monofunctional silanes, trifunctional silane (C18H37SiH3) yielded surfaces that showed good hydrolytic stability. This suggests that the stability of the monolayers from trifunctional silanes is primarily due to "horizontal" bonding (Si-O-Si or Si OH...HO-Si) rather than due to bondingwith the matrix (M-O-Si). At 65 degrees C, all C18 surfaces become more susceptible to hydrolysis; however, the trend observed for 25 degrees C remained unchanged. Low-temperature nitrogen adsorption was used to study the adsorption properties of the monolayers as a function of their grafting density. The energy of adsorption interactions showed a significant increase as the grafting density of the monolayers decreased. The order of the alkyl groups in the monolayers, as assessed from CH2 stretching, decreased as the grafting density of the monolayers decreased. PMID- 15835683 TI - Effects of the surface pressure on the formation of Langmuir-Blodgett monolayer of nanoparticles. AB - The effects of the surface pressure on the particle arrangement of Langmuir Blodgett (LB) monolayers of alkanethiol-capped gold nanoparticles were studied. The LB monolayers were prepared from a highly concentrated particle solution, which increases film fabrication efficiency but readily causes small particle voids in the particle array. Overcompressing the LB monolayer to a high surface pressure restructured the particles and eliminated the voids. When the gold particles capped by dodecanethiol were 8.5 nm in diameter, the particle arrangement was vastly improved and a wafer-scale LB monolayer was transferred onto a substrate at the surface pressure of 20 mN/m. PMID- 15835684 TI - Modeling drop shape on contaminated surfaces or surfaces with physical structures. AB - Surface contaminants are commonly found on films. They get transferred to the surface from incompletely cured silicone liners on the films or owing to migration of additives to the surface from within the film. During the process of ink jet printing (a noncontact printing process), surface contamination affects the shape of the drops (causing the formation of fingers and crescents) and hence image quality. This study uses modeling methods to examine how such surface contamination affects the drops shapes. Subsequently, it models the effect of surface structures (pits) on the drop shape. This study explores how image quality can be controlled in the presence of surface contamination and surface structures. PMID- 15835685 TI - Adsorption pattern of mixtures of trimethylammonium-modified hydroxyethylcellulose and sodium dodecyl sulfate at solid-liquid interfaces. AB - We studied mixtures of aqueous solutions of cationic hydroxyethylcellulose JR400 polymer and anionic sodium dodecyl sulfate using dynamic light scattering and atomic force microscopy (AFM). A ternary phase diagram was established showing three interesting realms of the polymer-surfactant-water mixture: a preprecipitation area of lowered viscosity (polymer excess) compared to the pure polymer solution, a postprecipitation area (resolubilization at surfactant excess), and highly diluted samples with a stoichiometrical surfactant-polymer ratio close to that of maximum precipitation. Samples with various compositions representing these areas were imaged by atomic force microscopy on mica and on hydrophobically modified silica in contact mode. A correlation between light scattering data concerning particle size and, more important, structuring in the bulk on one hand and AFM images on the other hand was observed. It was revealed that the influence of surface properties is of less importance for adsorption, compared to the influence of the mixture in the bulk, provided that the mixture is prepared prior to adsorption. PMID- 15835686 TI - Ultralow interfacial tensions in an aqueous phase-separated gelatin/dextran and gelatin/gum Arabic system: a comparison. AB - Many protein/polysaccharide mixtures phase separate when the concentrations ofthese biopolymers are sufficiently high. One of the properties involved in this phenomenon is the interfacial tension. Here we present measurements of the interfacial tension of two different protein/polysaccharide mixtures. The protein gelatin was mixed with either dextran or gum arabic, all used in a variety of food products. The phase diagrams were constructed using optical rotation. Although both polysaccharides have the same molecular weight, the phase diagrams differed. The interfacial tension of samples, varying in the distance from the critical point, was determined using the spinning drop method. The interfacial tension was found to be in the range of 1-15 microN/m. For both systems, the scaling behavior of the interfacial tension was investigated. The investigated gelatin/dextran system gave critical exponents of 2.5+/-0.1 and 1.4+/-0.1, in reasonable agreement with the mean-field values 3 and 1.5, respectively. The gelatin/gum arabic system did not show critical behavior. For this system, the interfacial tension shows a logarithmic dependence on the distribution of the gelatin and the gum arabic molecules in the separated phases. PMID- 15835687 TI - A small-angle neutron scattering study of adsorbed poly(ethylene oxide) on Laponite. AB - The adsorption of poly(ethylene oxide) (PEO) on synthetic anisotropic clay particles (Laponite) has been investigated as a function of the molecular weight. Contrast variation small-angle neutron scattering (SANS) measurements were used to characterize the distribution and adsorbed amount of polymer on the particles. These experiments show not only that polymer is present on the face of the clay particle but that it also extends or "wraps" over the edges. The edge layer was thicker than the face layer for all the molecular weights studied. The polymer layers are unusually thin, with a thickness and adsorbed amount that show little variation with molecular weight. PMID- 15835688 TI - Time-dependent changes in the formation of titania-based films at the air-water interface. AB - The growth of surfactant-assisted titanium dioxide-based films at the air-water interface previously reported (Henderson et al. Aust. J. Chem. 2003, 56, 933) has been monitored with a time resolution of minutes over the whole growth period by X-ray energy-dispersive reflectometry. Two new phenomena are described: (a) short term shifts in the Bragg spacing of the layer structure and (b) the periodic disappearance of the diffraction from the film. We associate these with changes in the chemistry of the reacting mixture, with changes in the packing of the templated titanium species, and with macroscopic and (possibly) microscopic rippling of the solid film during growth. PMID- 15835689 TI - Self-assembled molecular films of aminosilanes and their immobilization capacities. AB - The assembly of two aminosilanes on silicon dioxide surfaces is investigated in this work. It is found that for 3-aminopropyltrimethoxysilane (APS), a smaller concentration of the silane and trace amounts of water in the deposition medium, an optimum time, and a postdeposition thermal curing are necessary to obtain a high primary-amine content. By optimization of deposition conditions, uniform APS films with a primary-amine content of 88.6% were obtained. The dependence of the primary-amine content on the experimental parameters is related to the extent to which amines are lost to hydrogen bonding with each other or with the substrate surface. Whenp-aminophenyltrimethoxysilane (APhS) was used, the primary-amine content in the film reached 100% and the surface morphology was more uniform than that of APS films under the same conditions. This is attributed to the rigid phenyl component in APhS that reduces opportunities for hydrogen bonding. In a comparison of the immobilization capacities of the different aminosilane substrates for pyromellitic dianhydride (PMDA), it is observed that higher primary-amine content favors higher uptake, and the APhS film yields 100% PMDA coverage. We infer that primary-amine content could be a measure of the film morphology and accessibility of the substrate amine groups. PMID- 15835690 TI - On-line monitoring of polymer deposition for tailoring the waveguide characteristics of love-wave biosensors. AB - The Love-wave sensor is an acoustic sensing device which is particularly suitable for sensing in a liquid environment. The superior characteristics of the device are achieved by the use of an acoustic waveguide, consisting of a thin layer deposited on the surface of the substrate material. The exact thickness and material properties of the layer will not only determine sensitivity and sensing performance of the resulting device but can also be adjusted to generate higher order Love modes. Thus, to obtain a sensing device with the desired specifications, precise control over the process of waveguide deposition is required. This has been realized by implementation of a vapor deposition polymerization system where the transmission curve (amplitude vs frequency) of one of the sensing devices is continuously monitored during deposition. As soon as the desired device specifications are reached, the deposition can be interrupted immediately. From the recorded transmission curves, information about the sensitivity of the device can be deduced, and the formation of higher-order Love modes can be visualized. The system has been used to produce biosensors based on various Love modes. It is shown that sensors operated on higher-order Love modes have a high mass sensitivity which, together with their excellent shielding properties, makes them advantageous for biosensing in conducting buffer solutions. PMID- 15835691 TI - Dilatometric study of the adsorption of heavy-metal cations on goethite. AB - The specific volumes of adsorption of Cd, Co, Cu, Ni, Pb, and Zn on goethite determined by means of the dilatometric method are 21, 32, 32, 31, 31, and 42 cm3/mol, respectively, and are independent of pH. The effect of NaCl (up to 0.5 mol dm(-3)) on the specific volume of adsorption is rather insignificant. The specific volume of precipitation of corresponding hydroxides (determined experimentally and calculated) is about 60 cm3/mol. Apparently, the adsorbed heavy-metal cations lose half of their hydration water. The adsorption constant decreases as the pressure increases, and the effect becomes significant at pressures of > 10(7) Pa, i.e., more than 1 km of water column. PMID- 15835692 TI - Adsorption energy distribution from the Aranovich-Donohue lattice density functional theory. AB - We propose a new methodology projected for the estimation of the adsorption energy distribution from the monolayer part of a single nitrogen adsorption isotherm determined at 77 K based on the lattice density functional theory (DFT) via the Aranovich-Donohue formalism. At first sight, the presented approach is computationally more difficult than a classical one. However, it is more flexible and comprehensible. Next, we developed a numerical program and used it for the estimation of the adsorption energy distribution from the experimental data on carbon black samples. The main nitrogen molecule-carbon black surface interaction energy can be estimated as approximately 7-8 kJ/mol, but the heterogeneity of the investigated materials differs significantly. Furthermore, we compare the results obtained from the lattice DFT via the Aranovich-Donohue formalism with the solution of the integral equation with the kernel represented by the classical monolayer localized Fowler-Guggenheim isotherm equation. The similarity between these two independent approaches is observed. The proposed methodology can be used for the investigation of the energetic heterogeneity of not only the carbonaceous materials but also the other "flat-surfaced" solids. PMID- 15835693 TI - Attractive bridging interactions in dense polymer brushes in good solvent measured by atomic force microscopy. AB - Using an atomic force microscope (AFM), we have investigated the interaction forces exerted by latex particles bearing densely grafted polymer brushes consisting of poly(N,N-dimethylacrylamide) (PDMA), poly(methoxyethylacrylamide) (PMEA), poly(N-isopropylacrylamide) (PNIPAM), and PMEA-b-PNIPAM in aqueous media (good solvent). The brushes were prepared by controlled surface-initiated atom transfer radical polymerization, and the hydrodynamic thicknesses were measured by dynamic light scattering. The molecular weight (Mn), grafting density (sigma), and polydispersity (PDI) of the brushes were determined by gel permeation chromatography and multiangle laser light scattering after cleaving the polymer from the latex surface by hydrolysis. Force profiles of PDMA (0.017 nm(-2) < or = sigma < or = 0.17 nm-2) and PMEA (sigma = 0.054 nm-2) brushes were purely repulsive upon compression, with forces increasing with Mn and a, as expected, due to excluded volume interactions. At a sufficiently low grafting density (sigma = 0.012 nm-2), PDMA exhibited a long-range exponentially increasing attractive force followed by repulsion upon further compression. The long-range attractive force is believed to be due to bridging between the free chain ends and the AFM tip. The PNIPAM brush exhibited a bridging force at a grafting density of 0.037 nm(-2), a value lower than the sigma needed to induce bridging in the PDMA brush. Bridging was therefore found to depend on grafting density as well as on the nature of the monomer. The grafting densities of these polymers were larger than those typically associated with bridging. Bridging interactions were used to confirm the presence of PNIPAM in a block copolymer PMEA-b-PNIPAMA brush given that the original PMEA homopolymer brush produced a purely repulsive force. The attractive force was first detected in the block copolymer brush at a separation that increased with the length of the PNIPAM block. PMID- 15835694 TI - Kinetics of octadecyltrimethylammonium bromide self-assembled monolayer growth at mica from an aqueous solution. AB - We have studied the growth kinetics of self-assembled monolayers (SAMs) ofoctadecyltrimethylammonium bromide (C18TAB) on mica below the critical micelle concentration at 22, 30, 40, and 50 degrees C. A combination of atomic force microscopy, contact angle goniometry, and transmission infrared spectroscopy was used to follow the growth processes to determine the rates involved in the growth of a C18TAB SAM on mica. The growth of a SAM consisted of four distinct processes: deposition of adsorbate molecules, growth of a disordered 2D liquid phase, nucleation of islands of an ordered 2D solid phase, and subsequent growth of the solid phase. The rates of these various processes are determined, and the activation energies for several processes were calculated including those for the adsorption onto a bare substrate (20 kJ/mol), adsorption into the saturated liquid phase (100 kJ/mol), and nucleation of islands (0.3 kJ/mol). Despite the small activation barrier to island nucleation, the nucleation rate is qualitatively slow, suggesting that entropic effects dominate the nucleation rate. PMID- 15835695 TI - Microgravimetric study of electrochemically controlled nucleophilic addition of sulfite to polyaniline. AB - The sulfonation of polyaniline (PANI) films by nucleophilic addition of sulfite ion has been controlled through the polymer oxidation state under electrochemical control. The process was monitored by in situ electrochemical quartz crystal microbalance (EQCM), and the polymer oxidation was accomplished by electrode potential steps in sulfite aqueous solutions. The nucleophilic addition of sulfite to PANI only takes place on the oxidized polymer. From the ratio of added mass to the injected charge, the degree of sulfonation has been obtained with a yield as high as 50%. It has been observed that the ion-exchange mechanism during the oxidation-reduction process in the resulting sulfonated polymer is analogous to the polymer produced by electrophilic sulfonation of polyaniline or by copolymerization of aniline with aminosulfonic acids, unlike the ionic exchange observed for unmodified PANI. PMID- 15835696 TI - Roughness-induced acoustic second-harmonic generation during electrochemical metal deposition on the quartz-crystal microbalance. AB - This paper reports on the relation between the surface roughness and emission of compressional waves from the surface of an electrochemical quartz-crystal microbalance. The detection of the compressional waves took place with an ultrasonic microphone and the quartz crystal itself. As a model process, the electrochemical deposition of copper from an acidic copper sulfate solution has been chosen. For this system, the roughness of the layer can be tuned via the current density. Roughness may be a source of the longitudinal waves at twice the frequency of the exciting shear wave (acoustic second-harmonic generation, ASHG) if the flow profile above the quartz-crystal surface is not entirely laminar. Slight deviations from the laminar flow can be reached at high amplitudes of oscillation. Comparing the ASHG efficiency of a rough and smooth surface, we find that the rough surface is more efficient in generating second-harmonic waves. This suggests that ASHG can be used to obtain a roughness parameter independent from the resonance frequency or bandwidth (damping) of a quartz-crystal resonator. Such an independent determination of roughness should be very interesting in practical applications. PMID- 15835697 TI - Influence of the adsorption of N species on the anodic dissolution of Ni. AB - The dissolution and passivation of Ni in nitrite-containing acid solutions are investigated by Auger spectroscopy, atomic force microscopy, and conventional electrochemical techniques. The dissolution/ passivation of the Ni surface is consistent with a competition between adsorbed OH- and nitrogen-containing species with a potential-dependent surface coverage. Nitrogen-containing species hinder the passivation of the Ni surface, shifting the formation of the complex nickel hydroxide/oxide film to more positive potential values. The dynamics of the dissolving interface, followed by atomic force microscopy, reflect first the competition of adsorbed species, leading to the development of protrusions and cavities, and finally the formation of the passive film that promotes surface smoothening by a preferential dissolution of the protrusion tips under ohmic control. PMID- 15835698 TI - Ionic strength-dependent pK shift in the helix-coil transition of grafted poly(L glutamic acid) layers analyzed by electrokinetic and ellipsometric measurements. AB - Surface-bound layers of poly(L-glutamic acid) prepared by a recently described "grafting-from" method were analyzed with respect to electrical charging and structural alterations upon variation of pH and concentration of the background electrolyte in aqueous solutions. The microslit electrokinetic setup (MES) was utilized for the combined determination of zeta potential and surface conductivity on the basis of streaming potential and streaming current measurements at polypeptide layers in contact with aqueous electrolyte solutions of varied composition. In situ ellipsometry was applied at similar samples immersed in identical aqueous solutions to investigate the influence of the solution pH on the structure of the polypeptide layers. Zeta potential and Dukhin number versus pH plots revealed the dissociation behavior of the surface-bound polypeptides indicating a significant shift of the pK of their acidic side chains correlating with the concentration of the background electrolyte potassium chloride and the related variation of the Debye screening length. Surface conductivity data pointed at a more expanded structure of the polypeptide layer in the fully dissociated state as an increased ion conductance in this part of the interface was determined. The occurrence of a strong increase of the thickness and a corresponding decrease of the refractive index for the coil state of the layer strongly supports the findings of the electrokinetic measurements. This fully reversible "switching" of the layer structure was attributed to helix coil transitions within the grafted polypeptides induced by the dissociation of carboxylic acid functions of the polypeptide side chains. The shift of the "switching pH" of the surface-bound poly(L-glutamic acid) layers at varied concentrations of the background electrolyte was interpreted as a result of the pK shift of the carboxylic acid groups of the polypeptide side chains. The observed patterns prove that the electrostatic interactions causing this shift occur within but not between the grafted chains. PMID- 15835699 TI - Orientational behavior of thermotropic liquid crystals on surfaces presenting electrostatically bound vesicular stomatitis virus. AB - We report the orientational behavior of nematic phases of 4-cyano-4' pentylbiphenyl (5CB) on cationic, anionic, and nonionic surfaces before and after contact of these surfaces with solutions containing the negatively charged vesicular stomatitis virus (VSV). The surfaces were prepared on evaporated films of gold by either adsorption of poly-L-lysine (cationic) or formation of self assembled monolayers (SAMs) from HS(CH2)2SO3- (anionic) or HS(CH2)11(OCH2CH2)4OH (nonionic). Prior to treatment with virus, we measured the initial orientation of 5CB (delta epsilon = epsilon(parallel) - epsilon(perpendicular) > 0) to be parallel to the cationic surfaces (planar anchoring) but perpendicular (homeotropic) after equilibration for 5 days. A similar transition from planar to homeotropic orientation of 5CB was observed on the anionic surfaces. Only planar orientations of 5CB were observed on the nonionic surfaces. Because N-(4 methoxybenzylidene)-4-butylaniline (MBBA, delta epsilon = epsilon(parallel) - epsilon(perpendicular) < 0) exhibited planar alignment on all surfaces, the time dependent alignment of 5CB on the ionic surfaces is consistent with a dipolar coupling between the 5CB and electrical double layers formed at the ionic interfaces. Treatment ofpoly-L-lysine-coated gold films (cationic) with purified solutions of VSV containing 10(8)-10(10) plaque-forming units per milliliter (pfu/mL) led to the homeotropic alignment of 5CB immediately after contact of 5CB with the surface. In contrast, treatment of anionic surfaces and nonionic surfaces with solutions of VSV containing approximately 10(10) pfu/mL did not cause immediate homeotropic alignment of 5CB. These results and others suggest that homeotropic alignment of 5CB on cationic surfaces treated with VSV of titer > or = 10(8) pfu/mL reflects the presence of virus electrostatically bound to these surfaces. PMID- 15835700 TI - Complexes of poly(ethylene glycol)-based cationic random copolymer and calf thymus DNA: a complete biophysical characterization. AB - Complete biophysical characterization of complexes (polyplexes) of cationic polymers and DNA is needed to understand the mechanism underlying nonviral therapeutic gene transfer. In this article, we propose a new series of synthesized random cationic polymers (RCPs) from methoxy poly(ethylene glycol) monomethacrylate (MePEGMA) and (3-(methacryloylamino)propyl)trimethylammonium chloride with different mole ratios (32:68, 11:89, and 6:94) which could be used as a model system to address and answer the basic questions relating to the mechanism of the interaction of calf thymus DNA (CT-DNA) and cationic polymers. The solubility of the complexes of CT-DNA and RCP was followed by turbidity measurements. It has been observed that complexes of RCP with 68 mol % MePEGMA precipitate near the charge neutralization point, whereas complexes of the other two polymers are water-soluble and stable at all compositions. Dnase 1 digestion experiments show that DNA is inaccessible when it forms complexes with RCP. Ethidium bromide exclusion and gel electrophoretic mobility show that both polymers are capable of binding with CT-DNA. Atomic force microscopy images in conjunction with light scattering experiments showed that the complexes are spherical in nature and 75-100 nm in diameter. Circular dichroism spectroscopy studies indicated that the secondary structure of DNA in the complexes is not perturbed due to the presence of poly(ethylene glycol) segments in the polymer. Furthermore, we used a combination of spectroscopic and calorimetric techniques to determine complete thermodynamic profiles accompanying the helix-coil transition of CT-DNA in the complexes. UV and differential scanning calorimetry melting experiments revealed that DNA in the complexes is more stable than in the free state and the extent of stability depends on the polymer composition. Isothermal titration calorimetry experiments showed that the binding of these RCPs to CT-DNA is associated with small exothermic enthalpy changes. A complete thermodynamic profile showed that the RCP/DNA complex formation is entropically favorable. Much broader opportunities to vary the architecture of the polymers studied here make these systems promising in addressing various basic and practical problems in gene delivery systems. PMID- 15835701 TI - Collective and single-molecule interactions of alpha5beta1 integrins. AB - A novel biomimetic system was used to study collective and single-molecule interactions of the alpha5beta1 receptor-GRGDSP ligand system with an atomic force microscope (AFM). Bioartificial membranes, which display peptides that mimic the cell adhesion domain of the extracellular matrix protein fibronectin, are constructed from peptide-amphiphiles. The interaction measured with the immobilized alpha5beta1 integrins and GRGDSP peptide-amphiphiles is specifically related to the integrin-peptide binding. It is affected by divalent cations in a way that accurately mimics the adhesion function of the alpha5beta1 receptor. The recognition of the immobilized receptor was significantly increased for a surface that presented both the primary recognition site (GRGDSP) and the synergy site (PHSRN) compared to the adhesion measured with surfaces that displayed only the GRGDSP peptide. At the collective level, the separation process of the receptor ligand pairs is a combination of multiple unbinding and stretching events that can accurately be described by the wormlike chain (WLC) model of polymer elasticity. In contrast, stretching was not observed at the single-molecule level. The dissociation of single alpha5beta1-GRGDSP pairs under loading rates of 1-305 nN/s revealed the presence of two activation energy barriers in the unbinding process. The high-strength regime above 59 nN/s maps the inner barrier at a distance of 0.09 nm along the direction of the force. Below 59 nN/s a low strength regime appears with an outer barrier at 2.77 nm and a much slower transition rate that defines the dissociation rate (off-rate) in the absence of force (k(off) degrees = 0.015 s(-1)). PMID- 15835702 TI - How plants keep dry: a physicist's point of view. AB - This work describes the investigation of the physical basis of the amazing water repellence of some plant leaves, which is in addition to self-cleaning properties known as the "Lotus effect". Two rather different possible mechanisms are proposed, which are suggested to cover the main physics in a majority of superhydrophobic systems. These concepts are illustrated with two different types of plant leaves as well as a model surface carrying carbon nanotube aggregates. PMID- 15835703 TI - Interfacial and molecular properties of high-pressure-treated beta-lactoglobulin B. AB - Interfacial properties of beta-lactoglobulin B subjected to hydrostatic pressures up to 400 MPa were studied by measuring surface pressure at the air/water interface and the elastic interfacial shear modulus at the oil/water interface. The surface hydrophobicity of pressurized beta-lactoglobulin was determined by an 1-anilino-naphthalene-8-sulfonate assay and exposure of free thiol groups using the Ellman assay. The molar mass of pressure-induced oligomers was measured using a combination of size exclusion chromatography, light scattering, and refractive index measurements. High-pressure treatment of beta-lactoglobulin increased the surface pressure growth rate and its final level at the air/water interface. After high-pressure treatment, the maximum interfacial elasticity at the oil/water interface increased, and the time lag before growth of the interfacial elasticity decreased. Up to 200 MPa, large amounts of monomeric beta lactoglobulin were formed with increased exposure of thiol groups and increased surface hydrophobicity compared to unpressurized beta-lactoglobulin. At a pressure higher than 200 MPa, surface hydrophobicity continued to increase, while exposure of thiol groups decreased, the latter due to the formation of covalently linked oligomers. We have shown that surface hydrophobicity rather than thiol exposure is important for the pressure-induced increase in growth rate and the final level of surface pressure at the air/water interface and in interfacial elasticity at the oil/water interface. PMID- 15835704 TI - Engineered lipids that cross-link the inner and outer leaflets of lipid bilayers. AB - The application of supported lipid bilayer systems as molecular sensors, diagnostic devices, and medical implants is limited by their lack of stability. In an effort to enhance the stability of supported lipid bilayers, three pairs of phosphatidylcholine lipids were designed to cross-link at the termini of their 2 position acyl chain upon the formation of lipid bilayers. The cross-linked lipids span the lipid bilayer, resembling naturally occurring bolaamphiphiles that stabilize archaebacterial membranes against high temperatures. The three reactions investigated here include the acyl chain cross-linking between thiol and bromine groups, thiol and acryloyl groups, and cyclopentadiene and acryloyl groups. All three reactive lipid pairs were found to cross-link in liposomal membranes, as determined by thin-layer chromatography, ion-spray mass spectrometry, and 1H NMR. The monolayer film properties of the reactive amphiphiles were characterized by surface pressure-area isotherms and showed that stable monolayers formed at the air-water interface with limiting molecular areas comparable to that of pure saturated phosphatidylcholine lipids. Langmuir Blodgett bilayers of dimyristoylphosphatidylcholine incorporating 15 mol % of the reactive thiol and acryloyl lipids had diffusion coefficients comparable with pure dimyristoylphosphatidylcholine, while bilayers with more than 25 mol % of the reactive lipids were immobile, suggesting that interleaflet cross-linking of the lipids inhibited membrane diffusion. Our results show that the reactive lipids can cross-link within a lipid bilayer and are suitable for assembling supported lipid bilayers using Langmuir-Blodgett deposition. By using terminally reactive amphiphiles to build up supported lipid bilayers with cross-linked leaflets, bolaamphiphiles can be incorporated into asymmetric solid supported membranes to increase their stability in biosensor and medical implant applications. PMID- 15835705 TI - Fluorescent assays to quantitate enzymatic activities yielding as end product an aqueous-insoluble indigo-blue dye. AB - Hydrolytic enzymes acting upon indoxyl-derivatized substrates produce a water insoluble indigo-blue dye. The generation of indigo dye works well in nonquantitative histochemical or diagnostic assays. For quantitative analyses however, the technique is unsuited. In this paper two fluorescent methods are described that permit quantitative measurement of sialidase/neuraminidase activity toward indoxyl-derivatized N-acetyl-neuraminic acid substrates. The first method is based upon the reaction of the sialic/ N-acetyl-neuraminic acid with pyridoxamine and Zn2+ to produce a fluorescent chelate. This method is not sialic acid-specific and could be used for the quantitation of alpha-oxo acids. The optimum conditions for sialic acid fluorescent chelation are described. The second method is based upon the fluorescence of the reaction intermediates, indoxyl- and indigo-white, by arresting their conversion to nonfluorescent indigo blue. This method is suitable for measuring any enzymatic activity toward indoxyl derivatized substrates. Enzyme kinetics derived for influenza viral neuraminidase using the two techniques are described in this paper. PMID- 15835706 TI - Bidisperse electrorheological fluids using hydrolyzed styrene-acrylonitrile copolymer particles: synergistic effect of mixed particle size. AB - Monodisperse micron-sized styrene-acrylonitrile copolymer (SAN) particles with three different sizes (about 5, 10, and 15 microm) were prepared by a two-step seeded polymerization and used for a study of bidisperse electrorheological (ER) suspensions. The effect of the particle size and the size-mixing fraction on ER properties was studied with varying the size of these monodisperse copolymer particles. When the two particle sizes were mixed, the suspension generally showed a decrease in the shear yield stress, reaching a minimum value. However, a bidisperse ER suspension of large particles containing a small fraction of fine particles showed an interesting synergy effect of size mixing on ER response, giving enhanced yield stresses over the other size-mixing fractions. This synergistic ER suspension also showed a great increase in the viscoelastic property. The current density of suspensions was maximum at the synergistic bidisperse suspension. This synergy effect in a particular bidisperse suspension was investigated in view of the structure model consideration and was concluded to be due to a close packing and a peculiar structural ordering at an optimum size ratio and mixing fraction. PMID- 15835707 TI - Nanoparticle composition of a ferrofluid and its effects on the magnetic properties. AB - Experiments were carried out on a water-based ferrofluid (gamma-Fe2O3 with carboxydextran shell) using photon correlation spectroscopy (PCS), atomic force microscopy, and magnetic nanoparticle relaxation measurements. The experiments were designed with the aim to relate the Neel signals that are in theory generated by large single core particles with nanoscopic properties, that is, particle size, particle size distribution, shell properties, and aggregation. For this purpose, the ferrofluid was fractionated by magnetic fractionation and size exclusion chromatography. Nanoparticles adsorbed onto positively charged substrates form a two-dimensional monolayer. Their mean core diameters are in the range of 6 to about 20 nm, and particles above 10 nm are mostly aggregates. The hydrodynamic particle diameters are between 13 and 80 nm. The core diameter of the smallest fraction is confirmed by X-ray reflectometry; the surface coverage is controlled by bulk diffusion. Comparison with the hydrodynamic radius yields a shell thickness of 3.8 nm. Considering the shell thickness to be constant for all particles, it was possible to calculate the apparent particle diameter in the original ferrofluid from the PCS signals of all fractions. As expected, the small cores yielded no Neel relaxation signals in freeze-dried samples; however, the fractions containing mostly aggregates yielded Neel relaxation signals. PMID- 15835708 TI - Sub-100 nm patterning with an amorphous fluoropolymer mold. AB - A fluoropolymer mold is introduced and used to pattern sub-100 nm features with the characteristics that cause problems in patterning with a mold. The low surface energy and inertness, stiffness, and permeable nature of the mold material make it possible to pattern without surface treatment densely populated very fine features, mixed patterns of small and large features, and features with a high aspect ratio, when the mold is used with a polymer solution for the patterning. The ultraviolet transparency of the mold material also allows for patterning with photocurable pre-polymers. PMID- 15835709 TI - Temperature-sensitive hairy particles prepared by living radical graft polymerization. AB - Four types of temperature-sensitive hairy particles were prepared by living radical graft polymerization using a photoiniferter. The hairs were poly(N isopropylacrylamide) (N), poly(N-isopropylacrylamide)ran-poly(acrylic acid) (NA), and diblock copolymers composed of N and NA. The block copolymer was attached to the particle in different modes, that is, one has a N-block inner and a NA-block outer but the other has the inverse arrangement. The acrylic acid content in NA was adjusted to be only 1%, but NA had a higher transition temperature by 5 degrees C than N in a neutral aqueous solution. The sequence of blocks attached onto the particle was the key factor to control the temperature responsiveness of the particle. The hairy particles exhibited a two-step transition with increasing temperature under certain conditions. The hairy particle also responded to the pH and ionic strength. Some unique behaviors of the hairy particles were studied in detail in terms of electrophoretic mobility and adsorption of dye molecules as well as swelling/deswelling. PMID- 15835710 TI - Gelation of PEO-PLGA-PEO triblock copolymers induced by macroscopic phase separation. AB - The gelation behavior of aqueous solutions of poly(ethylene oxide-b-(DL-lactic acid-co-glycolic acid)-b-ethylene oxide) (PEO-PLGA-PEO) triblock copolymer containing short hydrophilic PEO end blocks is investigated using dynamic light scattering, rheology, small-angle neutron scattering (SANS), and differential scanning calorimetry (DSC). For polymer concentrations between 5 and 35 wt %, four distinct regions of the turbidity change depending on temperature were observed. Interestingly, in the turbid solution region, gel phase is formed for polymer concentrations above 14 wt % and an extremely slow relaxation was detected. In fact, a power law, which takes into account the dynamics of percolation clusters, dominates the correlation function. In rheological measurements, the local maximum in G' is observed at around the temperature of maximum turbidity. We further found that G" > G' and G' is highly dependent on frequency at the gel state implying viscoelastic characteristics, which is quite different from general concepts of gels, typically formed by the micellar packing. SANS profiles showing multiple peaks in the sol state rather than in the gel state as well as a DSC exotherm at the temperature of gels can also serve as the evidence of different gel states. Based upon the experimental data obtained in the present study, a new gelation mechanism induced by the macroscopic phase separation of triblock copolymers containing short hydrophilic PEO end blocks such as PEO-PLGA-PEO is proposed. The effect of the type ofhydrophobic middle blocks on the gelation is also discussed. PMID- 15835711 TI - Synthesis of TiO2 nanoparticles utilizing hydrated reverse micelles in CO2. AB - Titanium dioxide nanoparticles were produced by the controlled hydrolysis of titanium tetraisopropoxide (TTIP) in the presence of reverse micelles formed in CO2 with the surfactants ammonium carboxylate perfluoropolyether (PFPECOO-+NH4) (Mw = 587) and poly(dimethyl amino ethyl methacrylate-block-1H,1H,2H,2H perfluorooctyl methacrylate) (PDMAEMA-b-PFOMA). Based on dynamic light scattering measurements, the amorphous TiO2 particles formed by injection of TTIP are larger than the reverse micelles, indicating surfactant reorganization. The size of the particles and the stability of dispersions in CO2 were affected by the molar ratio of water to surfactant headgroup (w(o)), precursor concentration, and injection rate. The amorphous particle size did not change upon depressurization and redispersion in CO2. PDMAEMA-b-PFOMA provided greater stability against particle aggregation at higher reactant concentration compared with PFPECOO-+NH4. The crystallite size after calcination, which was examined by X-ray diffraction and transmission electron microscopy, increased with w(o). PMID- 15835712 TI - Superparamagnetism of magnetite nanoparticles: dependence on surface modification. AB - Superparamagnetic iron oxide nanoparticles (SPION) with an average particle diameter of 6 nm are prepared by controlled chemical coprecipitations. Colloidal suspensions of noninteracting SPION, where the surface has been modified with three different types of biocompatible substances, namely, starch, gold (Au), and methoxypoly(ethylene glycol) (MPEG) have been fabricated via three different techniques. Starch-coated SPION are prepared by coprecipitation in a polymeric matrix, Au-coated SPION are fabricated by the microemulsion method, and MPEG coated SPION are prepared using the self-assembly approach. The magnetic nanoparticles form a core-shell structure, and the magnetic dipole-dipole interactions are screened by a layer of coating agents. The amounts of coating agents and SPION are indirectly calculated from the thermogravimetric analysis and superconducting quantum interference device measurements by assuming passive oxidation on the surface of the SPION, and the other conditions do not influence the measurements. The dependency of the spectral characteristics of Mossbauer spectroscopy as a function of an external magnetic field Hext is measured to investigate the effect of dipole-dipole screening of the different coating layers on the SPION. Uncoated SPION show a stable magnetic moment under Hext, and the superparamagnetic (SPM) fraction transforms to a ferrimagnetic state. Starch and Au-coated SPION retain the SPM fraction according to Mossbauer spectroscopy and magnetization measurements. MPEG-coated SPION show hyperfine magnetic structure without the quadrupole effect with increasing the value of the blocking temperature. PMID- 15835713 TI - Patterned and controlled polyelectrolyte fractal growth and aggregations. AB - Two-dimensional patterned and controlled polyelectrolyte aggregations (e.g., tree like ramified structures) created by microcontact printing have been demonstrated and discussed. Polyelectrolyte-micropatterned aggregations on surfaces were controlled by the micropattern size and shape of PDMS stamps. The formation of aggregates was dependent on the ink and surface conditions, and the aggregates consisted of two distinct layers; strongly adsorbed, primary uniform layers and weakly adsorbed, secondary aggregation layers positioned on top of the primary layers. The adsorption of the primary layers was strong enough not to be washed away, while the aggregated secondary layers were easily removed by washing. The aggregation of secondary layers showed typical tree-like ramified structures of fractal growth and aggregation. Directional and confined stamping led to directing and confining the growth of the fractal polyelectrolyte clusters, respectively. The micropatterned primary uniform layers were not removed by extensive washing, and they were identified by selective nickel plating and charged particle selective adsorption in which the surface formed positive and negative micropatterns. These functional and patterned surfaces have great potentials for advanced devices and sensors. PMID- 15835714 TI - Hot filament chemical vapor deposition of poly(glycidyl methacrylate) thin films using tert-butyl peroxide as an initiator. AB - We have demonstrated the successful deposition of poly(glycidyl methacrylate) (PGMA) thin films using hot filament chemical vapor deposition (HFCVD) with tert butyl peroxide as the initiator. The introduction of the initiator allows for film deposition at low filament temperatures (<200 degrees C) and greatly improves the film deposition rates. The retention of the pendant epoxide chemical functionality and the linear polymeric structure in the deposited films were confirmed by infrared spectroscopy and X-ray photoelectron spectroscopy. The number-average molecular weight of the PGMA films can be systematically varied from 16,000 to 33,000 by adjusting the filament temperature and flow ratio of the initiator to the precursor. The apparent activation energies observed from PGMA deposition kinetics (100.9+/-9.6 kJ/mol) and from molecular weight measurements ( 54.8+/-2.0 kJ/mol) are close to the calculated overall activation energies for the polymerization rate (104.4 kJ/mol) and number-average molecular weight (-59.2 kJ/mol), which supports the hypothesis of the free radical polymerization mechanism in the HFCVD PGMA deposition. PMID- 15835715 TI - Effects of molecular siting and adsorbent heterogeneity on the ideality of adsorption equilibria. AB - The ideal adsorbed solution (IAS) theory is the benchmark for the prediction of mixed-gas adsorption equilibria from pure-component isotherms. In this work, we use atomistic grand canonical Monte Carlo simulations to test the effects of molecular siting and adsorbent energetic heterogeneity on the applicability of the IAS theory. Pure-component isotherms generated by atomistic simulation are used to predict binary isobaric isotherms using the IAS theory. These predicted isotherms are compared with those obtained by a full atomistic simulation of the binary mixture. Binary mixtures of argon, methane, and CF4 in silicalite are found to obey IAS theory, while benzene/methane and cyclohexane/methane in silicalite are nonideal. The mixture of argon and CF4 is ideal despite the large difference in the sizes of the two species. This contradicts previous hypotheses in the literature, which state that mixtures of species of unequal size do not adsorb ideally. The nonideal behavior of the benzene/methane and cyclohexane/methane systems occurs because of adsorbent heterogeneity in these systems, which depends on both sorbent and sorbate. In addition, we use a lattice gas model with parameters derived from atomistic simulation to demonstrate analytically that a sufficiently energetically heterogeneous adsorbent will result in the breakdown of IAS theory even in the absence of interactions between sorbates. PMID- 15835716 TI - Computer simulation of selective aggregation in binary colloids. AB - The morphology of clusters formed by selective aggregation of binary colloids is studied in a two-dimensional Monte Carlo simulation for a large range of number fractions (200:1, 100:1, 10:1, 2:1). We find remarkable similarity in morphology to those observed in experiments, from the formation of closed "micelles" to large branched clusters. Quantitative studies of the fractal dimension, kinetics, and cluster size distribution are also carried out and compared with diffusion limited cluster aggregation and reaction-limited cluster aggregation models. PMID- 15835717 TI - Modeling of the adsorption kinetics of surfactants at the liquid-fluid interface of a pendant drop. AB - This paper presents a theoretical model for simulating the adsorption kinetics of a surfactant at the liquid-fluid interface of a pendant drop. The diffusion equation is solved numerically by applying the semidiscrete Galerkin finite element method to obtain the time-dependent surfactant concentration distributions inside the pendant drop and inside the syringe needle that is used to form the pendant drop. With the obtained bulk surfactant concentration distributions, the adsorption at the interface is determined by using the conservation law of mass. It should be noted that the theoretical model developed in this study considers the actual geometry of the pendant drop, the depletion process of the surfactant inside the pendant drop, and the mass transfer of the surfactant from the syringe needle to the pendant drop. The present pendant-drop model is applied to study the adsorption kinetics of surfactant C10E8 (octaethylene glycol mono n-decyl ether) at the water-air interface of a pendant drop. The numerical results show that the Ward and Tordai equation, which was derived for adsorption from a semi-infinite surfactant solution to a planar interface, is unsuitable for interpreting the dynamic surface or interfacial tension data measured by using the pendant-drop-shape techniques, especially at low initial surfactant concentrations. The spherical-drop model, which assumes the pendant drop to be a perfectly spherical drop with the same drop volume, can be used to interpret the dynamic surface or interfacial tension data for pendant drops either with high initial surfactant concentrations or with low initial surfactant concentrations in short adsorption durations only. For pendant drops with low initial surfactant concentrations in long adsorption durations, the theoretical model developed in this study is strongly recommended. PMID- 15835718 TI - Temperature-sensitive critical micelle transition of sodium octanoate. PMID- 15835719 TI - Formation of porous films and vesicular fibers via self-organization of an amphiphilic chiral oligomer. PMID- 15835720 TI - The status of allopurinol in the management of tumor lysis syndrome: a clinical review. PMID- 15835721 TI - Retymicin, galtamycin B, saquayamycin Z and ribofuranosyllumichrome, novel secondary metabolites from Micromonospora sp. Tu 6368. I. Taxonomy, fermentation, isolation and biological activities. AB - A new xanthone compound named retymicin (1) was isolated together with galtamycin B (2) and saquayamycin Z (3), new members of the galtamycin and saquayamycin families, respectively, and the new lumichrome derivative 1-(alpha-ribofuranosyl) lumichrome (4) from Micromonospora strain Tu 6368, isolated from a soil sample collected in Romania. Retymicin, galtamycin B and saquayamycin Z show cytostatic effects to various human tumor cell lines whereas saquayamycin Z is also active against Gram-positive bacteria. PMID- 15835722 TI - Retymicin, galtamycin B, saquayamycin Z and ribofuranosyllumichrome, novel secondary metabolites from Micromonospora sp. Tu 6368. II. Structure elucidation. AB - A detailed screening of the secondary metabolite pattern from Micromonospora sp. strain Tu 6368 resulted in the isolation of ten compounds belonging to five different structural families. The structures of the novel compounds 1-(alpha ribofuranosyl)-lumichrome (3), retymicin (7), galtamycin B (11) and saquayamycin Z (14) were assigned by spectroscopic methods and chemical transformations. This strain fits our hypothesis that the metabolite analysis of biosynthetically talented strains leads readily to novel compounds. PMID- 15835723 TI - Sulphostin, a novel inhibitor of dipeptidyl peptidases IV (DPPIV) that stimulates hematopoiesis in mice. AB - CD26, a membrane-bound ectopeptidase, is known as an activated T cell marker with dipeptidyl peptidase IV (DPPIV) activity that has diverse functional roles in the regulation of peptide hormones, neuropeptides, chemokines and growth factors. We recently isolated a novel inhibitor of DPPIV, sulphostin, from culture broth of Streptomyces sp. MK251-43F3. We investigated herein the hematopoietic effect of sulphostin in mice and found that sulphostin induced the production of granulocyte colony-stimulating factor (G-CSF), stimulated myeloblasts in bone marrow, and increased neutrophil numbers in peripheral blood in both normal mice and mice with cyclophosphamide-induced leucopenia. Sulphostin desulfonate, in addition to sulphostin, has a similar inhibitory effect on DPPIV and stimulatory effect on neutrophils. These results suggest that DPPIV/CD26 might be a novel target for hematopoietic stimulation and DPPIV inhibitors including sulphostin and derivatives may be candidates for further development. PMID- 15835724 TI - SM-216601, a novel parenteral 1beta-methylcarbapenem: structure-activity relationships of antibacterial activity and neurotoxicity in mice. AB - It has been reported that 2-(4-substituted thiazol-2-ylthio)-1beta-methyl carbapenems exhibit potent activity against methicillin-resistant staphylococci (MRS) and vancomycin-resistant enterococci (VRE). In order to develop a novel broad-spectrum carbapenem, the structure-activity relationships of a series of 2 (4-tetrahydropyridinylthiazol-2-ylthio)-1beta-methylcarbapenems and 4 dihydropyrrolyl thiazole analogs were investigated with regard to their activity against Gram-positive and especially Gram-negative bacteria and also their convulsant activity, which is a major side effect concern of carbapenems. The introduction of substituent(s) on the dihydropyrrole moiety did not cause remarkable changes in anti-MRS and VRE activities, but tended to lower the anti Gram-negative bacterial activity except in some cases of methyl group introduction. These substitutions did however cause a reduction of the convulsant activity, which was affected by the size and also the configuration of the substituent. In the case of SM-216601 (6), introduction of a methyl group brought about significant reduction in neurotoxicity while maintaining favorable anti Gram-negative bacterial activity. PMID- 15835725 TI - Bioactive cyclic peptides from the psychrotolerant fungus Penicillium algidum. AB - A new cyclic nitropeptide, psychrophilin D (1), together with two known cyclic peptides, cycloaspeptide A (2) and cycloaspeptide D (3), were isolated from the psychrotolerant fungus Penicillium algidum using C18 flash chromatography, LH-20 Sephadex and preparative HPLC. The structure of psychrophilin D (1) was derived from mass spectrometric information, 1D and 2D NMR spectra and Marfey's method. The compounds were tested in antimicrobial, antiviral, anticancer and antiplasmodial assays. Psychrophilin D (1) exhibited a moderate activity (ID50 10.1 microg/ml) in the P388 murine leukaemia cell assay. Cycloaspeptide A (2) and D (3) exhibited moderate activity (IC50 3.5 and 4.7 microg/ml, respectively) against Plasmodium falciparum. PMID- 15835726 TI - Clonostachysins A and B, new anti-dinoflagellate cyclic peptides from a marine derived fungus. AB - The two new anti-dinoflagellates, clonostachysins A and B, were obtained from a marine sponge derived fungus Clonostachys rogersoniana strain HJK9. Their chemical structures were determined by spectroscopic studies as highly N methylated cyclic peptides of the nine amino acids. The absolute stereochemistry was elucidated by the advanced Marfey's method. Both clonostachysins A and B exhibited a selectively inhibitory effect on a dinoflagellate Prorocentrum micans at 30 microM, but had no effect on other microalgae and bacteria even at 100 microM. PMID- 15835727 TI - Sch 486058: a novel cyclic peptide of actinomycete origin. AB - The structure of a novel cyclic peptide (1) produced by Actinomycete sp. has been assigned on the basis of extensive NMR and mass spectral data. PMID- 15835728 TI - Thermodynamic origin of the chiral recognition of tryptophan on teicoplanin and teicoplanin aglycone stationary phases. AB - The D-, L-tryptophan binding and the chiral recognition properties of the teicoplanin and teicoplanin aglycone (TAG) chiral stationary phase (CSPs) were compared at various column temperatures. The solute adsorption isotherms (bi Langmuir model) were determined for both the two CSPs using the perturbation method. It was demonstrated that the sugar units were involved in the reduction of the apparent enantioselectivity through two phenomena: (i) the inhibition of some enantioselective contacts with low-affinity binding regions of the aglycone and (ii) a decrease in the stereoselective properties of the aglycone high affinity binding pocket. The phenomenon (ii) was governed by both a decrease in the ratio of the enantiomer adsorption constant and a strong reduction of the site accessibility for D- and L-tryptophan. In addition, a temperature effect study was performed to investigate the chiral recognition mechanism at the aglycone high-affinity pocket. An enthalpy-entropy compensation analysis derived from the Grunwald model as well as the comparison with the literature data demonstrated that the enantioselective binding mode was dependent on an interface dehydration process. The change in the enantioselective process observed between the TAG and teicoplanin CSP was characterized by a difference of ca. 2-3 ordered water molecules released from the species interface. PMID- 15835729 TI - Preparation and evaluation of a doubly tethered chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid. AB - A new doubly tethered chiral stationary phase (CSP) was prepared to enhance CSP stability as well as to take advantage of the tertiary amide linkage by bonding (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid to bis(3-aminopropyl)silica gel. The new CSP was quite effective in the resolution of alpha-amino acids, beta amino acids, amino alcohols, and amines and the chromatographic resolution behaviors of the new CSP were exactly consistent with those of the corresponding singly tethered CSP containing N-CH3 tertiary amide linkage. Direct comparison between the stabilities of the new CSP and the corresponding singly tethered CSP containing N-CH3 tertiary amide linkage under harsh chromatographic conditions reveals that the new CSP is more stable than the latter. PMID- 15835730 TI - Enantiomeric resolution of new aromatase inhibitors by liquid chromatography on cellulose chiral stationary phases. AB - Analytical HPLC methods using derivatized cellulose chiral stationary phases were developed for the direct enantioseparation of substituted [1-(imidazo-1-yl)-1 phenylmethyl)]-benzothiazolinone and benzoxazolinone derivatives with one chiral center. Those analogues of fadrozole constitute new potent nonsteroidal inhibitors of aromatase (P450 arom). The separations were made using normal phase methodology with a mobile phase consisting of n-hexane-alcohol (ethanol, 1 propanol, or 2-propanol) in various proportions, and a silica-based cellulose tris-3,5-dimethylphenylcarbamate (Chiralcel OD-H), or tris-methylbenzoate (Chiralcel OJ). The effects of concentration of various aliphatic alcohols in the mobile phase were studied. A better separation was achieved on cellulose carbamate phase compared with the cellulose ester phase. The effects of structural features of the solutes along with the temperature of the column on the discrimination between the enantiomers were examined. Baseline separation (Rs > 1.5) was easily obtained in many cases. PMID- 15835731 TI - Characterisation of poly(N-isopropylacrylamide) by asymmetrical flow field-flow fractionation, dynamic light scattering, and size exclusion chromatography. AB - Asymmetrical flow field-flow fractionation (AsFIFFF) was used to determine the hydrodynamic particle sizes, molar masses, and phase transition behaviour of various poly(N-isopropylacrylamide) (PNIPAM) samples synthesised by reversible addition--fragmentation chain transfer (RAFT) and conventional free radical polymerisation processes. The results were compared with corresponding data obtained by dynamic light scattering (DLS) and size exclusion chromatography (SEC). Agreement between the three methods was good except at higher molar masses, where the molar mass averages obtained by SEC were much lower than those obtained by AsFIFFF and light scattering. The aggregation of the polymers, which are thermally sensitive, was studied by DLS and AsFIFFF at various temperatures. In deionised water there was an abrupt change in the particle size due to phase separation at approximately equal to 32-35 degrees C. The critical temperatures determined by AsFIFFF were 3-5 degrees C higher than those obtained by DLS. PMID- 15835732 TI - LC-PDA and LC-ESI-MS separation and determination of process-related substances arising from stilbene-type fluorescent whitening agents. Application to monitoring of their photodegradation products in industrial effluents and aqueous environmental systems. AB - A simple and rapid gradient elution high-performance liquid chromatographic method using photodiode array and electrospray ionization mass spectrometric detectors was developed for separation and determination of the process-related substances and photodegradation products of stilbenesulfonic acids, viz. 4,4' dinitrostilbene-2,2'-disulfonic acid (DNSDA), 4-amino-4'-nitrostilbene-2,2' disulfonic acid (ANSDA), and 4,4'-diaminostilbene-2,2'-disulfonic acid (DASDA) in industrial waste waters. Gradient elution was carried out using ammonium acetate and acetonitrile as mobile phase and an Inertsil-ODS 3V column for separation. The negative-ion electrospray ionization mass spectra containing [M-H]- ions of sulfonic acids allowed molecular mass determination of unknowns and the structures were proposed on the basis of the fragment ions in the MS/MS spectra. PMID- 15835733 TI - Development and characterisation of a molecularly imprinted polymer prepared by precipitation polymerisation for the determination of phenylurea herbicides. AB - New materials based on molecularly imprinted polymers (MIPs) have been developed for use as sorbents in solid phase extraction to preconcentrate some urea herbicides. In the preconcentration step, different molecularly imprinted polymers were tested using methacrylic acid (MAA) and 2-(trifluoromethyl)acrylic acid (TFMAA) as functional monomers, and linuron and isoproturon as templates. The best results were obtained when the polymer was synthesised using MAA with isoproturon as template. Another parameter evaluated was the way in which the polymer was obtained. We observed that the imprinted polymers obtained by precipitation displayed a greater capacity to retain the phenylureas. Studies conducted using scanning electron microscopy (SEM) revealed that the bulk polymerisation method is far from ideal owing to the random shape and size distribution of the particles obtained, whereas when polymerisation was carried out in precipitation microspheres were obtained. In order to confirm the interaction between the functional monomer and the template, 1H NMR (CD2Cl2) analyses were conducted. The results obtained suggest that the hydrogen and/or nitrogen of the amino group of the template would be involved in the formation of hydrogen bonds with the functional monomer. The imprinted polymer obtained by precipitation polymerisation with MAA as functional monomer and isoproturon as template can be applied to preconcentrate phenylureas when the sample is dissolved in toluene. The proposed methodology was employed to evaluate polymer selectivity towards humic acids and towards other herbicides. PMID- 15835734 TI - Thioacetamide chemically immobilized on silica gel as a solid phase extractant for the extraction and preconcentration of copper(II), lead(II), and cadmium(II). AB - Thioacetamide immobilized on silica gel was prepared via the Mannich reaction. The extraction and enrichment of copper(II), lead(II), and cadmium(II) ions from aqueous solutions has been investigated. Conditions for effective extraction are optimized with respect to different experimental parameters in both batch and column processes prior to their determination by flame atomic absorption spectrometry (FAAS). The optimum pH ranges for quantitative adsorption are 4.0 8.0, 2.0-7.0, and 5.0-10.0 for Pb(II), Cu(II), and Cd(II), respectively. Pb(II) and Cd(II) can be desorbed with 3 mol/L and 0.1 mol/L HCl/HNO3, and Cu(II) can be desorbed with 2.5% thiourea. The adsorption capacity of the matrix has been found to be 19.76, 16.35, and 12.50 mg/g for Pb(II), Cu(II), and Cd(II), respectively, with the preconcentration factor of approximately equal to 300 for Pb(II) and approximately equal to 200 for Cu(II) and Cd(II). Analytical utility is illustrated in real aqueous samples generated from distilled water, tap water, and river water samples. PMID- 15835735 TI - Ion chromatographic analysis of amines, alkanolamines, and associated anions in concrete. AB - In order to assess the effectiveness of applying penetrating corrosion inhibitors to the surface of reinforced concrete, it is necessary to devise accurate methods for their detection and quantification. In this paper, methods for ion chromatographic analysis of a variety of amines, alkanolamines, and associated anions, which may be used as corrosion inhibitors for steel reinforcement in concrete, are described. By careful adjustment of the conditions for analysis, these inhibitors were readily identified and quantified in concrete/cement pore solutions or digests. Characterisation of the cationic inhibitors, ethanolamine, quaternary methylammonium, dimethylethanolamine, cyclohexylamine, guanidine, and arginine, and the anionic inhibitors, nitrite, molybdate, acetate, benzoate, and azelate, was carried out conductimetrically. To enhance the sensitivity of detection, amperometry was used for the analysis of triethanolamine and low concentrations of ethanolamine. Ion chromatography was also used as a means of obtaining a distribution profile of the concentrations of inhibitor ions present throughout a concrete structure. PMID- 15835736 TI - Application of supplementary flow in comprehensive 2D liquid chromatography combining SEC and RPC. AB - A comprehensive two-dimensional HPLC system (SEC x RPC) was evaluated. Various model compounds with differing hydrophobicity (log D: -0.08 to 2.22) and size (MW: 194 to 66.0 x 10(3)) were used. In order to reduce the run time of the second dimension, and thereby optimize the number of runs per unit volume from the first dimension, short RPC columns were used (7.5 mm). This column size demanded a low concentration of methanol in the mobile phase from the first dimension, in order to avoid severe band broadening and solute loss. Secondary interactions on SEC make a high methanol concentration in the mobile phase a necessity. Up to 40% methanol was required to diminish non-ideal SEC behavior. These conditions were non-compatible with trapping of hydrophilic compounds on RPC. The use of a supplementary flow (0.1% TFA) mixed after the first dimension led to better peak shape and trapping of hydrophilic compounds in the second dimension. This demanded flow adjustment in SEC, which in turn caused performance improvement and an increase in analysis time, making more RPC separations possible. These factors contributed to a larger peak capacity. PMID- 15835737 TI - Protein interactions with model chromatographic stationary phases constructed using self-assembled monolayers. AB - Model surfaces representative of chromatographic stationary phases were developed by immobilising an homologous series (C2-C18) of n-alkylthiols, mixed monolayers of C4/C18 and thioalkanes with alcohol, carboxylic acid, amino and sulphonic acid terminal groups onto a flat, silver-coated glass surface using self-assembled monolayer (SAM) chemistry. The processes of adsorption and desorption of serum albumins onto the monolayer surfaces was monitored in real-time using surface plasmon resonance (SPR). Alkyl-terminated SAMs all showed a strong adsorption of bovine serum albumin which was largely independent of alkyl chain length, the ratio of mixed C4/C18 SAMs or the solution pH/ionic strength. The adsorption of human serum albumin to carboxylic and amine terminated SAMs was shown to be predominantly via non-electrostatic interactions (hydrophobic or hydrogen bonding). However, sulphonic acid terminated SAMs showed almost exclusively electrostatic interactions with human serum albumin. This preliminary work using self-assembled monolayer chemistry confirms the usefulness of well characterised SAMs surfaces for investigating protein adsorption and desorption onto/from model chromatography surfaces and gives some guidance for selecting appropriate functionalities to develop better surfaces for chromatography and electrophoresis. PMID- 15835738 TI - Compartmental modeling of transdermal iontophoretic transport II: in vivo model derivation and application. AB - PURPOSE: This study was aimed to develop a family of compartmental models to describe in a strictly quantitative manner the transdermal iontophoretic transport of drugs in vivo. The new models are based on previously proposed compartmental models for the transport in vitro. METHODS: The novel in vivo model considers two separate models to describe the input into the systemic circulation: a) constant input and b) time-variant input. Analogous to the in vitro models, the in vivo models contain four parameters: 1) kinetic lag time (tL), 2) steady-state flux during iontophoresis (JSS), 3) skin release rate constant (KR), and 4) passive flux in the post-iontophoretic period (Jpas). The elimination from the systemic circulation is described by a) the one-compartment and b) the two-compartment pharmacokinetic models. The models were applied to characterize the observed plasma concentration vs. time data following single dose iontophoretic delivery of growth hormone-releasing factor (GRF) and R apomorphine. Moreover, the models were also used to simulate the observed plasma concentration vs. time profiles following a two-dose transdermal iontophoretic administration of alniditan. RESULTS: The time-variant input models were superior to the constant input models and appropriately converged to the observed data of GRF and R-apomorphine allowing the estimation of JSS, KR, and Jpas. In most cases, the values of tL were negligible. The estimated JSS and the in vivo flux profiles of GRF and R-apomorphine were similar to those obtained using the deconvolution method. The two-dose iontophoretic transport of alniditan was properly simulated using the proposed time-variant input model indicating the utility of the model to predict and to simulate the drug transport by a multiple dose iontophoresis. Moreover, the use of the compartmental modeling approach to derive an in vitro-in vivo correlation for R-apomorphine was demonstrated. This approach was also used to identify the optimum in vitro model that closely mimics the in vivo iontophoretic transport of R-apomorphine. CONCLUSIONS: The developed in vivo models demonstrate their consistency and capability to describe the in vivo iontophoretic drug transport. This compartmental modeling approach provides a scientific basis to examine in vitro-in vivo correlations of drug transport by iontophoresis. PMID- 15835739 TI - Intravenous hydrophobic drug delivery: a porous particle formulation of paclitaxel (AI-850). AB - PURPOSE: To develop a rapidly dissolving porous particle formulation of paclitaxel without Cremophor EL that is appropriate for quick intravenous administration. METHODS: A rapidly dissolving porous particle formulation of paclitaxel (AI-850) was created using spray drying. AI-850 was compared to Taxol following intravenous administration in a rat pharmacokinetic study, a rat tissue distribution study, and a human xenograft mammary tumor (MDA-MB-435) model in nude mice. RESULTS: The volume of distribution and clearance for paclitaxel following intravenous bolus administration of AI-850 were 7-fold and 4-fold greater, respectively, than following intravenous bolus administration of Taxol. There were no significant differences between AI-850 and Taxol in tissue concentrations and tissue area under the curve (AUC) for the tissues examined. Nude mice implanted with mammary tumors showed improved tolerance of AI-850, enabling higher administrable does of paclitaxel, which resulted in improved efficacy as compared to Taxol administered at its maximum tolerated dose (MTD). CONCLUSIONS: The pharmacokinetic data indicate that paclitaxel in AI-850 has more rapid partitioning from the bloodstream into the tissue compartments than paclitaxel in Taxol. AI-850, administered as an intravenous injection, has been shown to have improved tolerance in rats and mice and improved efficacy in a tumor model in mice when compared to Taxol. PMID- 15835740 TI - A novel, lipid-free nanodispersion formulation of propofol and its characterization. AB - PURPOSE: Propofol is a widely used anesthetic agent with highly desirable fast "on" and "off" effects. It is currently formulated as lipid emulsions, which are known to support microbial growth. In this study, a novel, lipid-free nanodispersion formulation of propofol was characterized. METHODS: The formulation was evaluated for its physical and chemical stability, in vitro compatibility with red blood cells, and its antimicrobial effectiveness. In vivo pharmacokinetic and pharmacodynamic properties of the formulation were evaluated in rats. RESULTS: Our data suggest that this lipid-free formulation is physically and chemically stable. Compared to the commercial emulsion formulation Diprivan, it causes less hemolysis with red blood cells and has improved antimicrobial activity. In addition, the lipid-free formulation demonstrates similar pharmacological effects to Diprivan in rats. CONCLUSIONS: This novel, lipid-free formulation exhibits improved in vitro properties without compromising in vivo effects, therefore representing a promising new alternative for propofol. PMID- 15835741 TI - A scalable, extrusion-free method for efficient liposomal encapsulation of plasmid DNA. AB - PURPOSE: A fully scalable and extrusion-free method was developed to prepare rapidly and reproducibly stabilized plasmid lipid particles (SPLP) for nonviral, systemic gene therapy. METHODS: Liposomes encapsulating plasmid DNA were formed instantaneously by mixing lipids dissolved in ethanol with an aqueous solution of DNA in a controlled, stepwise manner. Combining DNA-buffer and lipid-ethanol flow streams in a T-shaped mixing chamber resulted in instantaneous dilution of ethanol below the concentration required to support lipid solubility. The resulting DNA-containing liposomes were further stabilized by a second stepwise dilution. RESULTS: Using this method, monodisperse vesicles were prepared with particle sizes less than 200 nm and DNA encapsulation efficiencies greater than 80%. In mice possessing Neuro 2a tumors, SPLP demonstrated a 13 h circulation half-life in vivo, good tumor accumulation and gene expression profiles similar to SPLP previously prepared by detergent dialysis. Cryo transmission electron microscopy analysis showed that SPLP prepared by stepwise ethanol dilution were a mixed population of unilamellar, bilamellar, and oligolamellar vesicles. Vesicles of similar lipid composition, prepared without DNA, were also <200 nm but were predominantly bilamellar with unusual elongated morphologies, suggesting that the plasmid particle affects the morphology of the encapsulating liposome. A similar approach was used to prepare neutral egg phosphatidylcholine:cholesterol (EPC:Chol) liposomes possessing a pH gradient, which was confirmed by the uptake of the lipophilic cation safranin O. CONCLUSIONS: This new method will enable the scale-up and manufacture of SPLP required for preclinical and clinical studies. Additionally, this method now allows for the acceleration of SPLP formulation development, enabling the rapid development and evaluation of novel carrier systems. PMID- 15835742 TI - Cross-linked small polyethylenimines: while still nontoxic, deliver DNA efficiently to mammalian cells in vitro and in vivo. AB - PURPOSE: Polyethylenimine (PEI) is among the most efficient nonviral gene delivery vectors. Its efficiency and cytotoxicity depend on molecular weight, with the 25-kDa PEI being most efficient but cytotoxic. Smaller PEIs are noncytotoxic but less efficient. Enhancement in gene delivery efficiency with minimal cytotoxicity by cross-linking of small PEIs via potentially biodegradable linkages was explored herein. The hypothesis was that cross-linking would raise the polycation's effective molecular weight and hence the transfection efficiency, while biodegradable linkages would undergo the intracellular breakdown after DNA delivery and hence not lead to cytotoxicity. Toward this goal, we carried out cross-linking of branched 2-kDa PEI and its 1:1 (w/w) mixture with a linear 423-Da PEI via ester- and/or amide-bearing linkages; the in vitro and in vivo gene delivery efficiency, as well as toxicity to mammalian cells, of the resultant cross-linked polycations were investigated. METHODS: The efficiency of the cross-linked PEIs in delivering in vitro a plasmid containing beta-galactosidase gene and their cytotoxicity were investigated in monkey kidney cells (COS-7). Dynamic light scattering was used to compare the relative DNA condensation efficiency of the unmodified and cross-linked PEIs. In vivo gene delivery efficiency was evaluated by intratracheal delivery in mice of the complexes of a luciferase-encoding plasmid and the PEIs and estimating the luciferase expression in the lungs. RESULTS: Cross-linking boosted the gene delivery efficiency of the small PEIs by 40- to 550-fold in vitro; the efficiency of the most potent conjugates even exceeded by an order of magnitude that of the branched 25-kDa PEI. Effective condensation of DNA was evident from the fact that the mean diameter of the complexes of the cross-linked PEIs was some 300 nm with a narrow size distribution, while the complexes of the unmodified small PEIs exhibited a mean size of >700 nm with a very broad size distribution. At concentrations where the 25-kDa PEI resulted in >95% cell death, the conjugates afforded nearly full cell viability. The cross-linked PEIs were 17 to 80 times more efficient than the unmodified ones in vivo; furthermore, their efficiencies were up to twice that of the 25-kDa PEI. CONCLUSIONS: Cross-linking of small PEIs with judiciously designed amide- and ester-bearing linkers boosts their gene delivery efficiency both in vitro and in vivo without increasing the cytotoxicity. The high efficiency is dependent on the nature of the linkages and the PEIs used. PMID- 15835744 TI - Bis(carbamoyloxymethyl) esters of 2',3'-dideoxyuridine 5'-monophosphate (ddUMP) as potential ddUMP prodrugs. AB - PURPOSE: We previously reported the synthesis of bis(pivaloyloxymethyl) 2',3' dideoxyuridine 5'-monophosphate (POM2-ddUMP) (1a) as a membrane-transport prodrug formulation of the free parent nucleotide, ddUMP. Although successful at delivering ddUMP into cells in culture, POM2-ddUMP was rapidly degraded by plasma carboxylate esterases after intravenous administration to experimental animals, and therefore has limited therapeutic potential as a systemically administered prodrug. We now report the synthesis of bis(N,N'-dimethylcarbamoyloxymethyl)- and bis(N-piperidinocarbamoyloxymethyl) 2',3'-dideoxyuridine 5'-monophosphate [DM2 ddUMP (1b) and DP2-ddUMP (1c), respectively], analogues of POM2-ddUMP that were designed to be more resistant to degradation by plasma esterases. METHODS: After entering cell by passive diffusion, it was anticipated that loss of one of the carbamoyloxymethyl groups of 1b and 1c would occur by spontaneous chemical hydrolysis to give the intermediate phosphodiesters, 2b and 2c. Cleavage of the remaining carbamoyloxymethyl groups by cellular phosphodiesterase I would generate ddUMP. 1b and 1c were prepared by condensation of 2',3'-dideoxyuridine (ddU) with the appropriate bis(N-alkylcarbamoyloxymethyl) phosphate in DMA in the presence of triphenylphosphine and diethyl azodicarboxylate (the Mitsunobo reagent). RESULTS: The half-lives of 1b and 1c when incubated at a concentration of 10(-4) M in human plasma at 37 degrees C were 3.5 h and 3.7 h, respectively, similar to the half-lives observed under the same temperature conditions in 0.05 M aqueous phosphate buffer, pH 7.4. By contrast, the half-life of the POM2 prodrug, 1a, in plasma was only 5 min. The initial products of degradation of 1b and 1c were the phosphodiesters 2b and 2c. The latter compounds gave rise to ddUMP when incubated with snake venom phosphodiesterase I. CONCLUSION: These findings support the premise inherent in the design of 1b and 1c, namely that the carbamate prodrugs are far more resistant to hydrolysis by plasma carboxylate esterases than their POM counterparts and can revert to the free parent 5' mononucletides by successive chemical and enzymatic hydrolysis. Further studies of 1b and 1c as membrane-permeable prodrugs of ddUMP are in progress. PMID- 15835743 TI - A new class of 5-fluoro-2'-deoxyuridine prodrugs conjugated with a tumor-homing cyclic peptide CNGRC by ester linkers: synthesis, reactivity, and tumor-cell selective cytotoxicity. AB - PURPOSE: Tumor-targeting prodrugs of 5-fluoro-2'-deoxyuridine (5-FdUrd), which are chemical conjugations of 5-FdUrd with a tumor-homing cyclic peptide CNGRC by succinate and glutarate linkers, were synthesized to investigate the structural effects of linkers on the hydrolytic release of 5-FdUrd and the tumor-cell selective cytotoxicity. METHODS: A solid phase synthesis method was used to produce 5-FdUrd prodrugs. The kinetics and efficiency of hydrolytic 5-FdUrd release from the prodrugs were investigated in phosphate buffer (PB), fetal bovine serum (FBS), HT-1080 cell lysate, MDA-MB-231 cell lysate, and MEM containing 10% FBS. The tumor-cell-selective cytotoxicity of prodrugs was evaluated by an MTT method. RESULTS: Two tumor-targeting prodrugs CNF1 and CNF2 bearing 5-FdUrd conjugated with a common cyclic peptide CNGRC by succinate and glutarate linkers, respectively, and their control compounds CN1 and CN2 without 5-FdUrd moiety were synthesized and identified. CNF1 underwent hydrolysis to release 5-FdUrd more rapidly and efficiently than CNF2. Both prodrugs were of lower cytotoxicity compared to 5-FdUrd, showing more selective cytotoxicity toward APN/CD13 positive cells (HT-1080) than toward APN/CD13 negative cells (HT 29, MDA-MB-231). CONCLUSIONS: A new class of tumor-targeting 5-FdUrd prodrugs CNF1 and CNF2 were successfully synthesized. These prodrugs targeted a tumor marker APN/CD13 to cause tumor-cell-selective cyctotoxicity due to 5-FdUrd release, the rate of which could be controlled by the structure of ester linker. PMID- 15835745 TI - Significant intestinal excretion, one source of variability in pharmacokinetics of COL-3, a chemically modified tetracycline. AB - PURPOSE: This study was undertaken to examine the disposition of COL-3, a chemically modified tetracycline, in order to elucidate its major route of elimination as one possible source of the variability in pharmacokinetics of COL 3 in vivo. METHODS: The disposition profile of COL-3 in vivo was assessed by examining the urinary and fecal excretion of the unchanged drug and/or its metabolites in rats after single intravenous and oral administration. The biliary excretion of COL-3 administered orally in bile duct-cannulated rats was also examined. In addition, plasma protein binding and cytochromes P450-mediated metabolism were explored along with erythrocyte partitioning in vivo. Furthermore, transport of COL-3 across Caco-2 monolayers was performed to elucidate the mechanism of intestinal excretion of COL-3 in vivo. RESULTS: COL-3 was extensively bound to plasma protein in rat (98%) and human plasma (95%). The affinity of rat blood cells for COL-3, as measured by the ratio of drug concentration in blood cells to that unbound in plasma, was about 36. Of the single intravenous and oral doses, less than 0.2% and 0.03% were excreted unchanged in rat urine, respectively; while 32.1 +/- 9.9% and 38.8 +/- 6.1% were recovered unchanged in rat feces, respectively, within 48 h postdosing. Of the oral dose, 1.36 +/- 0.66% and 2.97 +/- 0.88% were excreted in rat bile as the unchanged COL-3 and the total of COL-3 and its glucuronide conjugate, respectively, within 24 h after dosing. COL-3 had insignificant cytochrome P450 mediated metabolism but underwent phase II metabolism (i.e., glucuronidation) in a minor quantity. COL-3 was not a substrate of P-glycoprotein. Its transport across Caco-2 monolayers was significantly affected by protein binding and pH. CONCLUSIONS: Intestinal excretion, a route different from biliary excretion, is the major route of elimination for COL-3 in rats. Variability in intestinal excretion, due to extreme variable intestinal contents (food and digestive fluids), could be one source of variability in COL-3 pharmacokinetics in vivo in addition to the dissolution rate-limited absorption. PMID- 15835746 TI - Acute changes in muscle blood flow and concomitant muscle damage after an intramuscular administration. AB - PURPOSE: The intramuscular route (IM) is widely used but commonly induces injection site muscle damage. This study investigates the hemodynamic changes in an acute lesion induced by the IM administration of propylene glycol (PG) in rabbits. METHODS: Control groups received 1, 2, or 3 ml of PG (IM). Others were pretreated with pancuronium, dantrolene, indomethacin, or SR140333 and then received 2 ml of PG. The muscle blood flow (MBF) was assessed using fluorescent microspheres before and at 15, 45, 60, 90 min, 3 and 6 h after IM administration. Different areas within the muscle damage were quantified. RESULTS: Muscle contractions as well as a transient but major MBF increase were observed at the injection site. All treatments reduced hyperemia by up to 81% (dantrolene, 15 min) at 15, 45, and 90 min (p < 0.05). MBF had returned to basal values in all groups at 6 h. The central necrotic area was not modified, but peripheral damage (8.0 +/- 1.3 g) was reduced by dantrolene, indomethacin, and SR140333 (p < 0.05), but not by pancuronium. CONCLUSIONS: Muscle contraction and hyperemia are not responsible for muscle damage at the injection site, which is the multifactorial phenomenon, involving intracellular calcium and inflammation. PMID- 15835747 TI - Relationship between loperamide-induced sedative effect and digoxin pharmacokinetics in healthy Japanese subjects. AB - PURPOSE: Loperamide-induced suppressive effects on central nervous system closely relate to a lack of or decline in the P-glycoprotein (P-gp) function. The aim of this study was to determine the loperamide-induced sedative effect quantitatively and to investigate possible alterations in the pharmacokinetics of digoxin, a substrate for P-gp, in Japanese subjects. METHODS: Loperamide hydrochloride (2 mg) was administered orally to 26 subjects and the critical flicker-fusion frequency threshold (CFF) values were measured every 30 min separately by portable instrument. Further, digoxin (0.25 mg) was administered to 8 subjects, and the plasma concentration was determined. RESULTS: In five subjects who complained of drowsiness, the CFF values more remarkably decreased compared with those in the other subjects. The Tmax and mean residence time (MRT) values of digoxin pharmacokinetics in four subjects with drowsiness were significantly lower and Cmax was higher than those in four subjects with marginal effect. Moreover, there were good correlations between the CFF value-time profile and the Cmax, Tmax, and MRT of digoxin. CONCLUSIONS: The determination of the CFF value after oral administration of loperamide will be useful for evaluating varied P-gp function and for anticipating individual variations in the disposition of P-gp substrates in humans. PMID- 15835748 TI - Pharmacokinetic and tissue distribution mechanism of mouse recombinant heat shock protein 70 in mice. AB - PURPOSE: To investigate the in vivo pharmacokinetics and uptake mechanisms of recombinant mouse heat shock protein 70 (Hsp70) by hepatocytes in mice. METHODS: The tissue distribution and intrahepatic localization of Hsp70 were determined after an intravenous injection of 111In-Hsp70 (111In-Hsp70) into mice. Ligands of CD91 or scavenger receptors were injected prior to Hsp70 to examine the involvement of these molecules on the distribution of 111In-Hsp70. The uptake of 111In-Hsp70 by primary mouse hepatocytes was also examined. RESULTS: After intravenous injection, 111In-Hsp70 was rapidly eliminated from the circulation and taken up mainly by the liver. The hepatic uptake was significantly inhibited by preinjection of ligands for CD91 or scavenger receptors. The separation of liver-constituting cells revealed a major contribution of hepatocytes to the overall hepatic uptake of 111In-Hsp70. The uptake of 111In-Hsp70 by cultured hepatocytes was inhibited by a CD91 ligand or anti-CD91 anibody. In addition, after subcutaneous injection, 111In-Hsp70 gradually disappeared from the injection site and accumulated in primary lymph nodes. CONCLUSIONS: These results indicate for the first time that intravenous Hsp70 is, at least partially, recognized by CD91 and eliminated by hepatocytes, whereas subcutaneous Hsp70 is efficiently delivered to regional lymph nodes. PMID- 15835749 TI - Formulation and evaluation of aerosolized celecoxib for the treatment of lung cancer. AB - PURPOSE: We examined the effect of aerosolized celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on the in vitro cytotoxicity and apoptotic response of docetaxel against the human lung carcinoma cell lines A549 and H460. METHODS: A metered dose inhaler (MDI) formulation of celecoxib was prepared and evaluated for its medication delivery and aerodynamic properties. The in vitro cytotoxicity of the aerosolized celecoxib-MDI alone or in combination with docetaxel was assessed using a six-stage viable impactor by a previously established method. The induction of apoptosis was evaluated by morphologic examination (acridine orange and Hoechst staining) and DNA fragmentation. Furthermore, in an attempt to identify molecular targets involved in the anticancer mechanisms of celecoxib and docetaxel, we examined their effect on the expression of an array of markers involved in the COX-2 dependent and independent pathways. RESULTS: The celecoxib-MDI had a medication delivery of 231.3 microg/shot, mass median aerodynamic diameter (MMAD) of 1.4 microm (GSD = 1.9), and respirable fraction of 50.7%. The celecoxib-MDI (2 shots) in combination with docetaxel had cell kills as high as 81.3% and 67.7% in A549 and H460 cells, respectively. Hoechst and acridine orange staining showed an enhanced induction of apoptosis in A549 and H460 cells exposed to aerosolized celecoxib with docetaxel, which was further confirmed by DNA fragmentation. Western blot analysis showed a significant reduction in cPLA2 expression in both A549 and H460 cells treated with the combination of celecoxib with docetaxel. In the COX-2 independent pathway, there was a significant increase in the expression of PPAR gamma and p53, whereas pro-caspase-3 expression was significantly decreased, which may contribute to the enhanced apoptotic response observed with the combination treatment. CONCLUSIONS: Our results suggest that aerosolized celecoxib significantly enhances the in vitro cytotoxicity and apoptotic response of docetaxel against A549 and H460 cells, and this enhanced activity is mediated via alterations in expression of various molecular targets involved in apoptosis. PMID- 15835750 TI - Phase behavior of amorphous molecular dispersions II: Role of hydrogen bonding in solid solubility and phase separation kinetics. AB - PURPOSE: To determine the factors influencing "solid solubility" and phase separation kinetics of drugs from amorphous solid dispersions. METHODS: Solid dispersions of griseofulvin-poly(vinyl pyrrolidone) (PVP) and indoprofen-PVP were prepared using solvent evaporation technique. Dispersions demonstrating single Tg were exposed to 40 degrees C/69% RH for 90 days. Drug solid solubility in the polymer and phase separation rates were determined from changes in Tg of solid dispersions. FTIR spectroscopy and XRD were used to characterize drug-polymer interactions and drug crystallinity, respectively. RESULTS: Freshly prepared solid dispersion of up to 30% w/w griseofulvin and indoprofen were molecularly miscible with PVP. Hydrogen bonding was evident in indoprofen-PVP, but not in griseofulvin-PVP dispersions. When exposed to 40 degrees C/69% RH, griseofulvin phase separated completely, whereas the solid solubility of indoprofen was determined as 13% w/w. The first-order rate constants of phase separation for 10%. 20%, and 30% w/w griseofulvin dispersions were estimated as 4.66, 5.19, and 12.50 (x10(2)) [day(-1)], and those of 20% and 30% w/w indoprofen dispersions were 0.62 and 1.25 (x10(2)) [day(-1)], respectively. CONCLUSIONS: Solid solubility of griseofulvin and indoprofen in PVP is approximately 0% w/w and approximately 13% w/w, respectively. Drug-polymer hydrogen bonding in indoprofen PVP dispersions favors solid solubility. Phase separation rate of drug from the solid dispersions depends on the initial drug content and the nature of drug polymer interactions. PMID- 15835751 TI - Isolated rafts from adriamycin-resistant P388 cells contain functional ATPases and provide an easy test system for P-glycoprotein-related activities. AB - PURPOSE: P-glycoprotein (P-gp), a membrane ATPase expelling many structurally unrelated compounds out of cells, is one of the major contributors to multidrug resistance. It is enriched in cold TritonX-100 insoluble membrane domains (i.e., rafts). The purpose of this work was to characterize the ATPase activities of raft preparations from P388 cells overexpressing P-gp (P388/ADR) or devoid of P gp (P388) and to establish a P-gp-enriched screening system for P-gp-interfering compounds. METHODS: Rafts were extracted with cold TritonX-100. The ATPase activity was characterized in 96-well plates using a fluorescence assay. RESULTS: The ATPase activity per mg protein was about five times higher in P388/ADR rafts than in crude membranes. The anti-P-gp antibody C219 inhibited 20% of the activity in P388/ADR rafts but only about 10% of the activity in P388/ADR crude membranes and had no effect on the activity of P388 rafts. The known P-gp activating compounds verapamil, progesterone, and valinomycin revealed the typical bell-shaped activity/concentration profiles in P388/ADR rafts, indicative for activation at low compound concentrations and inhibition at concentrations >10 to 100 microM. The inhibitory effect was also observed in P388 rafts. CONCLUSIONS: Extracted rafts are rich in functional ATPases. Rafts from P-gp overexpressing cells display P-gp-typical ATPase activity and provide an easy, P gp-enriched screening system. PMID- 15835752 TI - Characterization of polarized expression of point- or deletion-mutated human BCRP/ABCG2 in LLC-PK1 cells. AB - PURPOSE: In polarized cells, such as hepatocytes and intestinal epithelial cells, transporters are localized on the apical or basolateral membranes and play important roles in the vectorial transport of their substrates. In the current study, we have aimed to clarify the mechanism for the cellular sorting of human breast cancer resistance protein (BCRP/ABCG2), which is expressed on the apical membrane of many tissues and functions as an efflux transporter. METHODS: After the expression vector, including wild type or mutants of human BCRP cDNA, was transfected into LLC-PK1 cells, immunohistochemical staining and Western blot analyses were performed to characterize the cellular localization and the status of BCRP, respectively. RESULTS: The transfected cDNA product of wild-type BCRP was expressed on the apical membrane in LLC-PK1 cells. Glycosylation consensus sequences-disrupted mutants showed the apical localization as the wild type, whereas the apical-selective expression disappeared when disulfide bonds could not be formed. Furthermore, examination of the localization of deletion mutants of human BCRP emphasized the importance of some peptide sequences. The region between the N-terminal and ATP-binding cassette and proximal C-terminal region, both of which are well conserved in various animal species, were found to be significant for proper localization. CONCLUSIONS: These results suggest that, although the presence of N-glycan does not affect the localization of BCRP, disulfide bonds and some peptide sequences in both the N- and C-terminals are necessary for the apical expression of BCRP. PMID- 15835753 TI - Interaction of the macrolide antibiotic azithromycin with lipid bilayers: effect on membrane organization, fluidity, and permeability. AB - PURPOSE: To investigate the effect of a macrolide antibiotic, azithromycin, on the molecular organization of DPPC:DOPC, DPPE:DOPC, SM:DOPC, and SM:Chol:DOPC lipid vesicles as well as the effect of azithromycin on membrane fluidity and permeability. METHODS: The molecular organization of model membranes was characterized by atomic force microscopy (AFM), and the amount of azithromycin bound to lipid membranes was determined by equilibrium dialysis. The membrane fluidity and permeability were analyzed using fluorescence polarization studies and release of calcein-entrapped liposomes, respectively. RESULTS: In situ AFM images revealed that azithromycin leads to the erosion and disappearance of DPPC and DPPE gel domains, whereas no effect was noted on SM and SM:cholesterol domains. Although azithromycin did not alter the permeability of DPPC:DOPC, DPPE:DOPC, SM:DOPC, and SM:Chol:DOPC lipid vesicles, it increased the fluidity at the hydrophilic/hydrophobic interface in DPPC:DOPC and DPPE:DOPC models. This effect may be responsible for the ability of azithromycin to erode the DPPC and DPPE gel domains, as observed by AFM. CONCLUSIONS: This study shows the interest of both AFM and biophysical methods to characterize the drug-membrane interactions. PMID- 15835754 TI - Novel film modifiers to alter the physical properties of composite ethylcellulose films. AB - PURPOSE: Polyvinylpyrrolidone (PVP), molecular-composite PVP, and Plasdone S-630 copolyvidonum are potential polymeric film modifiers for achieving improved drug release. The aim of this study was to investigate how these polymeric additives would affect the physicomechanical properties of composite ethylcellulose films. METHODS: The miscibility of these polymeric additives with ethylcellulose was determined from the differential scanning calorimetry (DSC) thermograms of various polymer blends formed from organic solvents. It was found that ethylcellulose (EC) was miscible with the polymeric additives up to a concentration of 50%. Ten percent to 30% w/w polymeric additives were then added to aqueous ethylcellulose dispersion to form composite films. The morphology, film transparency, dynamic mechanical analysis (DMA) thermograms, and mechanical properties of the composite ethylcellulose films were studied. In addition, puncture strength and % elongation of the dry and wet films were also compared from indentation test. RESULTS: Significant reduction and change in film transparency and morphology was obtained for EC films blended with PVP of higher molecular weight (MW). The composite EC films also showed higher Tg, greater elastic modulus, tensile and puncture strength depending on the concentration and type of additives present. CONCLUSIONS: The interaction between ethylcellulose and the polymeric additives is dependent on the MW and concentration of additives. The composite films offer new opportunities for the use of ethyl cellulose as modified release coatings for dosage forms. PMID- 15835756 TI - [Age-related features of the brain bioelectrical activity formation in children with remote consequences of a perinatal lesion of the CNS: I. Spontaneous activity]. PMID- 15835755 TI - Comparative investigation by two analytical approaches of enthalpy relaxation for glassy glucose, sucrose, maltose, and trehalose. AB - PURPOSE: In an effort to understand the stability of glassy sugars such as glucose, sucrose, maltose, and trehalose, the molecular mobility below the glass transition temperature (Tg) was investigated by an enthalpy relaxation measurement with differential scanning calorimetry (DSC). METHODS: The glassy sample was aged over several days at (Tg - 10) K to (Tg - 30) K, before a DSC heating scan was taken. The relaxed enthalpy (deltaHrelax) was estimated from the endothermic peak area. The enthalpy relaxation time was analyzed from the time course of deltaHrelax using two different approaches; Kohlrausch-Williams-Watts (KWW) and extended Adam-Gibbs (exAG). RESULTS: TauKWW, which is defined as the mean average enthalpy relaxation time in a distribution, and tau(eff)0 and tau(eff)infinity, which correspond to the enthalpy relaxation time of the initial minimum and final maximum cooperative rearrangement region, were estimated by KWW and exAG, respectively. And three activation energies for enthalpy relaxation were calculated from the Arrhenius plot. CONCLUSIONS: Although these deltaEs originated from different theoretical backgrounds, almost the same trend was observed for a comparison of the values of the four sugars. The finding that the deltaEs of glassy trehalose were the largest among the four sugars may support the reason that glassy trehalose is an effective stabilizer. PMID- 15835757 TI - [Age-related changes in the mechanisms of image classification in young schoolchildren with different cognitive styles]. PMID- 15835758 TI - [Study of the genesis of pathological EEG patterns in tumor and traumatic lesions of the human brain]. PMID- 15835759 TI - [Spectral analysis of the heart rate, blood pressure, and respiratory rate variability in 8- to 11-year-old children at rest]. PMID- 15835760 TI - [Regularities of changes in microcirculatory blood flow through different parts of the heart in patients with acquired valvular disease before and after surgery]. PMID- 15835761 TI - [Bioelectrical cardiac processes during induced systemic vasodilation in men under conditions of the Russian North]. PMID- 15835762 TI - [Characteristics of the maintenance of the upright posture during additional contact with an external object on a moving or immobile platform]. PMID- 15835763 TI - [Manifestation of morphofunctional specificity of muscles in spectral evaluations of the integrated electromyogram]. PMID- 15835764 TI - [Parameters of psychophysiological adaptation to the academic process in the department of physical training in students specializing in different sports]. PMID- 15835765 TI - [Autonomic regulation of the heart rate in humans under conditions of acute experimental hypoxia]. PMID- 15835766 TI - [Saprotrophic bacteria as a possible means of controlling the immune status of the body through regulation of metabolic reactions]. PMID- 15835767 TI - [Physiology of chronic inflammatory processes and their treatment]. PMID- 15835768 TI - [Evaluation of the parameters of total, nutritive, and shunt blood flows in the skin microvasculature using laser doppler flowmetry]. PMID- 15835769 TI - [Gastric secretion, motility, and emptying in hyperkinesia]. PMID- 15835770 TI - [The role of I.P. Pavlov in the development of emergency cardiology]. PMID- 15835771 TI - [Contribution of alpha-adrenoceptors to the contractility of the human myocardium in chronic coronary insufficiency]. PMID- 15835772 TI - [Appearance and disappearance of motile ciliated epithelial cells in sputum during acute respiratory infection]. PMID- 15835773 TI - [Developing the control criterion for continuous culture of microorganisms]. AB - A short survey and critical analysis of previously proposed criteria for growth control of populations of microorganisms in the chemostat are presented. Based on the analysis of a mathematical model of the steady-state of a microbial population in the chemostat, an adequate control criterion is suggested, along with a method to identify the corresponding regulating factors. The new control criterion is expressed as a product of the factor transformation coefficient and the biomass sensitivity coefficient (SC) with respect to the change of the factor at the chemostat inlet (referred to in the sequel as the biomass SC). The control criterion determines the strength of the control exerted by this or that factor. The method of determination of the regulating factors consists in experimental determination of the real SCs for factors and the biomass and in calculating on this basis the corresponding ideal SCs for constant factor transformation coefficients. The ideal SCs are shown to add up to an integer value, a constraint that we call "quantization" relationships. Such relationships are used to test the completeness of the drawn list of control factors. The proposed method was applied to our own and literature data. PMID- 15835774 TI - [The functional role of reduced inorganic sulfur compounds in the metabolism of the microaerophilic bacterium Spirillum winogradskii]. AB - Oxidation of reduced sulfur compounds by microaerophilic sulfur bacterium Spirillum winogradskii was found to occur only concomitantly with consumption of an organic substrate and was not linked to their utilization as electron donors in energy metabolism. No enzymes of dissimilatory sulfur metabolism were found in the cells of the sulfur bacterium oxidizing thiosulfate to tetrathionate; oxidation of thiosulfate and sulfide was caused by their reaction with reactive oxygen species (ROS), mostly H2O2 produced in the course of aerobic growth. Decreased lytic effect of ROS in the presence of thiosulfate resulted in a twofold increase in the cell yield under aerobic conditions and more efficient substrate utilization. The latter effect was caused by decreased expense of energy for the biosynthesis of oxygen-protecting polysaccharides. The stimulatory effect of thiosulfate on the growth processes was due to the activation of a number of TCA cycle enzymes producing the intermediates for constructive metabolism, especially of the NADP-dependent malic enzyme. As a result of thiosulfate-induced synthesis of SH-containing cell components, the integral antioxidative activity increased 1.5-fold. PMID- 15835775 TI - [The role of alkylhydroxybenzenes in the adaptation of Micrococcus luteus to heat shock]. AB - The response of the gram-positive bacterium Micrococcus luteus to heat shock (45 degrees C, 15 min) and the adaptogenic activity of alkylhydroxybenzenes (AHB), which are extracellular growth-regulating substances of these bacteria, were studied. The perception of stress and the postshock behavior of M. luteus cells proved to depend on the growth phase and medium. The magnitude of stress response was more pronounced in cultures grown on synthetic medium than in cultures grown on rich medium (nutrient broth). During exponential or linear growth, the cells were more sensitive to the temperature effect than during decelerated growth. In linearly growing M. luteus cultures, the amount of total intra- and extracellular alkylhydroxybenzenes, the anabiosis inducers, increased in response to heat shock. AHB redistribution between cells and culture liquid occurred in the course of stress and after stress. In micrococci exposed to heat shock, an increase in the AHB concentration both in cells and culture liquid is likely a defense reaction of stress resistance. This conclusion was confirmed in the experiments with the addition 30 min before the heat shock of a chemical analogue of the anabiosis inducer, C7-AHB (12 mM), which protected M. luteus cells so that their intense growth was observed after shock without any lag. The protective effect of AHB is a result of their ability to form complexes with enzyme macromolecules and stabilize them. The data obtained extend the knowledge of the stress-protective functions of low-molecular-weight autoregulators and of the role of intercellular communications in the stress response of bacterial cultures. PMID- 15835776 TI - [Secretion of bacterial ribonuclease]. AB - The investigation of the effect of some components of the medium on the distribution of the secretory guanyl-specific ribonuclease of Bacillus intermedius (EC 3.1.4.23) among various cell fractions and culture liquid showed that the amount of this enzyme in the culture liquid does not depend on the concentration of calcium ions in the medium (within 1-5 mM). The study of the effect of the amino acid substitutions Trp34Asn and Trp70Asn in the ribonuclease molecule showed that the secretion of ribonuclease depends on the formation rate of its secondary structure. The amino acid substitution Trp34Asn completely inhibits ribonuclease secretion. PMID- 15835777 TI - [Biosynthesis of Bacillus intermedius glutamyl endopeptidase in recombinant Bacillus subtilis cells during the stationary growth phase]. AB - We studied the biosynthesis of Bacillus intermedius glutamyl endopeptidase in the recombinant Bacillus subtilis strain AJ73 delta58.21 during the stationary growth phase. We optimized the composition of the culture medium to favor effective enzyme production during the stationary growth phase, and found that the nutritional requirements for glutamyl endopeptidase synthesis were different in the stationary phase and growth retardation phase. Proteinase accumulation was activated by complex organic substrates (casein and gelatin). During final stages of the culture growth, the enzyme production was stimulated by Ca2+, Mn2+, and Co2+ and inhibited by Zn2+, Fe2+, and Cu2+. The synthesis of glutamyl endopeptidase in the late stationary phase was not inhibited by glucose, unlike that in the trophophase during proliferation. We conclude that the regulatory mechanisms of proteinase synthesis during vegetative growth and sporulation are different. PMID- 15835778 TI - [Cell wall teichoic acids from Streptomyces daghestanicus VKM Ac-1722T and Streptomyces murinus INA-00524T]. AB - The structure of cell wall teichoic acids was studied by chemical methods and NMR spectroscopy in the type strains of two actinomycete species of the "Streptomyces griseoviridis" phenetic cluster: Streptomyces daghestanicus and Streptomyces murinus. S. daghestanicus VKM Ac-1722T contained two polymers having a 1,5 poly(ribitol phosphate) structure. In one of them, the ribitol units had alpha rhamnopyranose and 3-O-methyl-alpha-rhamnopyranose substituents; in the other, each ribitol unit was carrying 2,4-ketal-bound pyruvic acid. Such polymers were earlier found in the cell walls of Streptomyces roseolus and Nocardiopsis albus, respectively; however, their simultaneous presence in the cell wall has never been reported. The cell wall teichoic acid of Streptomyces murinus INA-00524T was is a 1,5-poly(glucosylpolyol phosphate), whose repeating unit was [-6)-beta-D glucopyranosyl-(1 --> 2)-glycerol phosphate-(3-P-]. Such a teichoic acid was earlier found in Spirilliplanes yamanashiensis. The 13C NMR spectrum of this polymer is presented for the first time. The results of the present investigation, together with earlier published data, show that the type strains of four species of the "Streptomyces griseoviridis" phenetic cluster differ in the composition and structure of their teichoic acids; thus, teichoic acids may serve as chemotaxonomic markers of the species. PMID- 15835779 TI - [Auxin production by bacteria associated with orchid roots]. AB - Bacteria associated with the roots of greenhouse tropical orchids were shown to produce indole-3-acetic acid (IAA) and to excrete it into the culture liquid. The presence and activity of IAA were demonstrated colorimetrically, by thin-layer chromatography, and by biotests. The associated bacteria varied in their ability to excrete indole compounds (1-28 microg/ml nutrient broth). Addition of tryptophan to the growth medium enhanced phytohormone production. Upon addition of 200 microg/ml tryptophan, the bacteria isolated from Dendrobium moschatum roots (Sphingomonas sp. 18, Microbacterium sp. 23, Mycobacterium sp. 1, Bacillus sp. 3, and Rhizobium sp. 5) produced 50.2, 53.1, 92.9, 37.6, and 60.4 microg IAA/ml respectively, while the bacteria isolated from Acampe papillosa roots (Sphingomonas sp. 42, Rhodococcus sp. 37, Cellulomonas sp. 23, Pseudomonas sp. 24, and Micrococcus luteus) produced 69.4, 49.6, 53.9, 31.0, and 39.2 microg IAA/ml. Auxin production depended on cultivation conditions and on the growth phase of the bacterial cultures. Treatment of kidney bean cuttings with bacterial culture liquid promoted formation of a "root brush" with location height 7.4- to 13.4-fold greater than the one in the control samples. The ability of IAA producing associated bacteria to act as stimulants of the host plant root development is discussed. PMID- 15835780 TI - [The synthesis of hydroxybutyrate and hydroxyvalerate copolymers by the bacterium Ralstonia eutropha]. AB - The paper deals with the study of the synthesis of 3-hydroxybutyrate (3HB) and 3 hydroxyvalerate (3HV) copolymers by the bacterium Ralstonia eutropha B-5786 grown under different carbon nutrition conditions (growth on carbon dioxide, fructose, and CO2-valerate and fructose-valerate mixtures). The parameters to be analyzed included the yield of biomass, the yield, synthesis rate, and composition of copolymers, the activity of the key enzymes of polyhydroxyalkanoate (PHA) synthesis (beta-ketothiolase, acetoacetyl-CoA reductase, and PHA synthase), the maximum tolerable concentration of valerate to the bacterium, and the conditions that govern the incorporation of hydroxyvalerate to copolymers. This allowed the relationship between cultivation conditions and the proportion of monomers in the copolymers to be deduced. We were able to synthesize a range of 3HB/3HV copolymers and found that the thermal characteristics and the degree of crystallinity of these copolymers depend on the molar fraction of 3HV. PMID- 15835781 TI - [Diversity of genetic systems responsible for biodegradation of naphthalene in Pseudomonas fluorescens strains]. AB - The genetic systems that are responsible for naphthalene catabolism were analyzed in 18 naphthalene-degrading Pseudomonas fluorescens strains isolated from oil contaminated soils in different regions of Russia. It was found that thirteen strains contain plasmids, from 20 to 120 kb in size, at least five of which are conjugative and bear the catabolic genes responsible for the complete utilization of naphthalene and salicylate. Five plasmids belong to the P-7 incompatibility group, and two plasmids belong to the P-9 incompatibility group. The naphthalene biodegradation genes of P. fluorescens are highly homologous to each other. The study revealed a new group of the nahAc genes and two new variants of the nahG gene. The suggestion is made that the key genes of naphthalene biodegradation, nahAc and nahG, evolve independently and occur in P. fluorescens strains in different combinations. PMID- 15835782 TI - [Effect of transposons on expression of genes for naphthalene biodegradation in Pseudomonas putida BS202(NPL-1) and derivative strains]. AB - NPL-1 and its derivative plasmid pBS106, which control the degradation of naphthalene and salicylate, were found to contain class II transposons of the Tn3 family. These transposons are involved in intraplasmid rearrangements, such as deletions and inversions, and can influence the expression of the catabolic and regulatory genes borne by biodegradation plasmids. The formation of a strong NahR independent constitutive promoter by the inversion of a DNA fragment may be responsible for changing the character of naphthalene dioxygenase synthesis from inducible (in the case of plasmid NPL-1) to constitutive (in the case of plasmid NPL-41). The stability of plasmids NPL-1 and NPL-41 in the Pseudomonas putida strains grown on different substrates depends on the expression of the nah and tnp genes. PMID- 15835783 TI - [The chemotactic characteristics of the S and R dissociants of Bacillus thuringiensis]. AB - The chemotactic responses of Bacillus thuringiensis subsp. dendrolimus (strain 49) and thuringiensis (strain 2002) and their morphological dissociants were studied by using some natural and artificial substances as effectors. The 12-h old wild-type cells (S variants) of both strains were found to be motile and similar in their chemotactic responses, whereas the chemotactic responses of R variants were different. PMID- 15835784 TI - [Isolation, purification, and identification of virulence protein VirE2 from Agrobacterium tumefaciens]. AB - Bacteria of the genus Agrobacterium are capable of transferring a fragment of their Ti-plasmid, T-DNA, in a complex with the proteins VirE2 and VirD2, into the nuclei of plant cells and incorporating it into the chromosome of the host. The mechanisms of T-DNA transportation through membrane and cytoplasm of the plant cell are unknown. The aim of this work was isolation of virulence protein VirE2 for studying its role in T-DNA transportation through the membrane and cytoplasm of eukaryotic cells. For VirE2 accumulation, virE2 gene was cloned into plasmid pQE31. VirE2 was isolated from the cells of E. coli strain XL1-blue, containing the recombinant plasmid pQE31-virE2. The cells were disrupted ultrasonically, and the protein with six histidine residues at the N-end was isolated by means of affinity chromatography on a Ni-NTA-superose column. The purified protein was tested by the immunodot method using polyclonal rabbit antibodies and anti-VirE2 miniantibodies. The ability of the recombinant protein VirE2 to bind to single stranded DNA was judged from the formation of complexes detected by electrophoresis in agarose gel. Thus, we isolated, purified, and partially characterized the Agrobacterium tumefaciens virulence protein VirE2 which is capable of binding to single-stranded T-DNA upon transfer to the plant cell. PMID- 15835785 TI - [The prevalence of cis-9-hexadecenoic acid is a specific feature of the fatty acid profile of zygomycetes from the order Kickxellales]. AB - The fatty acid profiles of zygomycetes from the family Kickxellaceae of the order Kickxellales were studied with reference to the species Kicksella alabastrina of the key genus Kicksella of the family and the species Linderina pennispora. When synthesized de novo, the lipids of these species show the prevalence of cis-9 hexadecenoic acid. This trait is stable, does not depend on cultivation conditions, and can, therefore, be considered as a specific chemotaxonomic characteristic of fungi from the order Kickxellales. The fatty acid profiles of the two fungi under study are similar to that of sea buckthorn oil. PMID- 15835786 TI - [The structure of micromycete communities and their synecologic interactions with basidiomycetes during decomposition of plant debris]. AB - We investigated the interactions between micromycetes and basidiomycete mycelium on plant substrates in the course of their 3-year incubation in the litter of ecologically intact spruce forests of the Central State Biosphere Forest Sanctuary (Nelidovo District, Tver oblast). Only 40-60% of the micromycetes were involved in direct antagonistic interactions with basidiomycetous fungi. In terms of the ratio between physiologically active strains and those which did not interact with basidiomycete mycelium, we revealed differences in the structure of micromycete communities developing on various types of substrates (xylem, bark, sphagnum, leaves, needles, litter, and cotton grass). The micromycetes tested belonged to 49 species. At the end of the observation period, the fraction of microscopic fungi that actively influenced basidiomycete mycelium was four times lower in the inactive litter fraction (lignin-containing xylem debris) than in the active fraction (grass substrates). The mechanisms of indirect regulation of the structure and functions of micromycete communities are discussed, which may be based on the accumulation of phenolic compounds in the medium and changes in the enzyme activities of basidiomycete mycelium. PMID- 15835787 TI - [Microbiological processes in the Lost City vent field, mid-Atlantic ridge]. AB - Microbiological and biogeochemical measurements showed that the intensities of CO2 assimilation, methane oxidation, and sulfate reduction in the Lost City vent field (30 degrees N) reach 3.8 microg C/(1 day), 0.06 microg C/(1 day), and 117 microg S/(1 day), respectively. On the surface of the carbonate structures occurring in this field, two varieties of bacterial mats were found. The first variety, which is specific to the Lost City alkaline vent field, represents jelly bacterial mats dominated by slime-producing bacteria of several morphotypes. This mat variety also contains chemolithotrophic and heterotrophic microorganisms, either microaerobic or anaerobic. The intensities of CO2 assimilation, methane oxidation, and sulfate reduction in this variety reach 747 microg C/(dm3 day), 0.02 microg C/(dm3 day), and 28,000 microg S/(dm3 day), respectively. Bacterial mats of the second variety are formed by nonpigmented filamentous sulfur bacteria, which are close morphologically to Thiothrix. The intensities of CO2 assimilation, methane oxidation, and sulfate reduction in the second mat variety reach 8.2 microg C/(dm3 day), 5.8 microg C/(dm3 day), and 17,000 microg S/(dm3 day), respectively. These data suggest the existence of subsurface microflora in the Lost City vent field. PMID- 15835788 TI - [Butyric acid bacteria of the genus Clostridium in the bottom sediments of inland basins of different types]. AB - The cell numbers and ecological characteristics of the distribution of certain species of butyric acid bacteria (BAB) of the genus Clostridium in the bottom sediments of inland basins of different types were studied using the optimal nutrient media. The seasonal dynamics of clostridial vegetative cells and spores in sediments with different ecological conditions were revealed. The cell numbers of the dominant BAB species were shown to depend on the redox potential of the sediments, the amount and composition of C(org), and the trophic state of the basin in general. C. pasteurianum was found to predominate in eutrophic lakes and reservoirs (5-11 x 10(6) cells/cm3), C. butyricum and C. felsineum predominated in mesotrophic ones (2-11 x 10(6) cells/cm3), and C. acetobutylicum was predominant in acidic chthionoeutrophic lakes and reservoirs (0.1-0.5 x 10(6) cells/cm3). The lowest cell numbers of BAB were found in river sediments, whereas the highest numbers were recorded in the sediments of polysaprobic zones (0.1-1.0 x 10(3) and 0.5-2.0 x 10(7) cells/cm3 respectively). PMID- 15835789 TI - [The effect of ampicillin on the electrophysical properties of Escherichia coli cells]. AB - The study of the effect of ampicillin on the electrophysical properties of Escherichia coli cells showed that this antibiotic influences the orientational spectra (OS) of the ampicillin-susceptible E. coli strains K-12 and XL-1 within the frequency range 10-1000 kHz of the orienting electric field and does not affect the OS of the ampicillin-resistant strains K-12(pUC-18) and XL-1(pHEN1). The change in the electrooptical signal of the ampicillin-susceptible cells was maximum at an ampicillin concentration of 50 microg/ml and did not depend on the exposure time. The conclusion is drawn that changes in the OS of cells can be used to evaluate their resistance to ampicillin. PMID- 15835790 TI - [The conjugated transfer of plasmid pUB110 in Bacillus subtilis in soils from different natural landscapes]. PMID- 15835791 TI - [Dependence of the relative value of redox potential in Rhodococcus rhodochrous cells on their growth rate in culture]. PMID- 15835792 TI - [Mask lift and facial aesthetic sculpturing]. AB - OBJECTIVE: To improve the outcome of the face-lift with a craniofacial contouring procedure. METHODS: Thirty-seven patients aged 29-53 years (31 in female, 6 in male) were treated through a bicoronal incision. The facial tissue was lifted through a subperiosteal procedure and the facial skeleton was remodeled in three dimensions by osteotomies. RESULTS: The results were satisfactory after the 3-12 months of the follow-ups. CONCLUSIONS: The Face-lift combined with the facial aesthetic sculpturing may be more effective for the aging-face rejuvenating. PMID- 15835793 TI - [Aesthetic correct depressed nose and prolong nasal columella using nasal columella base flaps, bilateral labial mucosa flaps, and the silicone imitation]. AB - OBJECTIVE: Using simple method to reform depressed middle part of the face and to prolong nasal columella. METHODS: Fill silicone into the subcutaneous cavity of the nase and maxillary attended by using nasal columella base and bilateral labial mucosa flaps. RESULTS: Obtain a satisfactory result to reform depressed nose,nasal columella and maxilla. CONCLUSIONS: It is a propagable economic way to resolve a complex deformation using this simple method, and get a satisfactory result. PMID- 15835794 TI - [Long-term results after the correction of the paralytic ectropion caused with leprosy in 115 eyes]. AB - OBJECTIVE: To analyze the long-term results after the correction of the paralytic ectropion in leprosy. METHODS: Seventy-four patients with 115 paralytic ectropion eyes after leprosy were treated with the surgical procedures included medial canthoplasty, medial canthal tendon plication, lateral tarsal strip, medial canthal resection, lateral canthoplasty, and lid shortening. The results were evaluated with the follow-ups from 2 to 4 years. RESULTS: The eye-redness was reduced from 93 to 40 while the epiphora from 107 eyes (24 mild, 36 moderate and 47 severe) to 90 (40 mild, 32 moderate and 18 severe). The mean lid gap in mild eye closure was reduced from 6.8 mm to 5.3 mm and the cornea lesion was reduced from 53 to 36. The results were excellent in 18 eyes (15.7%), good in 45 eyes (39.1%), fair in 41 eyes (35.7%) and poor in 11 eyes (9.6%). However, the mean visual acuity remained same pre- and postoperatively. CONCLUSIONS: Surgical correction of ectropion is helpful for cornea protection and could improve the signs and symptoms of the eyes such as epiphora and red [abstract truncated]. PMID- 15835795 TI - [Therapy experience and pathogeny analysis of intercurrent infection after augmentation mammaplasty]. AB - OBJECTIVE: To investigate the pathogeny diagnosis, therapy and some possible method for prevention of intercurrent infection, after the operation of augmentation mammaplasty. METHODS: Comparing and analyzing the clinical representation and therapy of the seventeen intercurrent infecting case that we had, after the operation of the augmentation mammaplasty. The patients were treated by operating with different approaches. RESULTS: After the operation, all the infected patients recovered in a short while without back to surgery. CONCLUSIONS: The infection after augmentation mammaplasty will get an effective controlling result if take a timely operative therapy according to the position and character of the intercurrent infection. But some galactophore will shrink after the cure of infection. PMID- 15835796 TI - [The expression of the estrogen receptor within the mammary gland of the breast hypertrophy and hypogenesis]. AB - OBJECTIVE: To investigate the expression of the estrogen receptors(ER) in breast hypertrophy and micromastia. METHODS: The quantity of ER within breast tissue in 13 patients with mammary hypertrophy strictly elected was determined using DCC single dot test. The results were compared with that measured in 13 patients with micromastia. RESULTS: The average of the quantity of the ER was respectively (5.33 +/- 5.70) fmol/mg and (1.17 +/- 0.86) fmol/mg in mammary hypertrophy ground and in the micromastia group. There was obvious significant between the two groups in a statistics basis (P < 0.05) . CONCLUSIONS: The breast hypertrophy is related to the excessive expression of ER within breast tissue,while the micromastia and breast hypotrophy are related to the lower level or the expression inhibition of ER. PMID- 15835797 TI - [Penile augmentation using acellular dermal matrix]. AB - OBJECTIVE: Penile enhancement was performed using acellular dermal matrix. METHODS: Multiple layers of acellular dermal matrix were placed underneath the penile skin to enlarge its girth. Since March 2002, penile augmentation has been performed on 12 cases using acellular dermal matrix. RESULTS: Postoperatively all the patients had a 1.3-3.1 cm (2.6 cm in average) increase in penile girth in a flaccid state. The penis had normal appearance and feeling without contour deformities. All patients gained sexual ability 3 months after the operation. One had a delayed wound healing due to tight dressing, which was repaired with a scrotal skin flap. CONCLUSIONS: Penile enlargement by implantation of multiple layers of acellular dermal matrix was a safe and effective operation. This method can be performed in an outpatient ambulatory setting. The advantages of the acellular dermal matrix over the autogenous dermal fat grafts are elimination of donor site injury and scar and significant shortening of operation time. PMID- 15835798 TI - [Study of genic mutations of androgen receptor in hypospadias]. AB - OBJECTIVE: To study genic mutation of androgen receptor in hypospadias in Chinese. METHODS: Ninety-two patients with hypospadias were selected for the AR gene study (penile 33, scrotal 49, perineal 10; cryptorchidism 15, micropenis 34, indirect hernia 3, augmentation of breast 2, other deformities 8; posses family history 23) while 93 health men as the control. The DNA was collected from 5 ml venous blood by using the method of phenol/chloroform, and quantified with the agarose gel electrophoresis. The 2-8 exons of AR genes were directly sequenced with a PCR technique. The locus of mutation,the change of amino acid caused by genic mutation and the functional influence of target organ were also analyzed. RESULTS: There were 4 mutations found from the ligand binding domain of AR: exon 4[1 locus:664(ATT-->ACT] and exon 7[3 locuses: 840(CGT-->CAT),855(CGC- >CAC),859(CTC-->CTA)] in experimental group. The amino acid change of Iso664Thr may te the first findings in the world. The mutation of 859(CTC-->CTA) didn't cause the change of the code amino acid. The main clinical symptoms of exon 4[664(ATT-->ACT) were micropenis, slight hypospadias and augmentation of breasts. The exon 7[840(CGT-->CAT), 855(CGC-->CAC)]was showing micropenis and severe hypospadias. The last mutation [859(CTC-->CTA)] of exon 7 showed mild hypospadias. CONCLUSIONS: The rate of exon 2-7 mutations of the androgen receptor was 4.3%. The mutations were concentrated on ligand binding domain(exon 4-7), and the main phenotypes were hypospadias accompanying with micropenis found in our experimental group. PMID- 15835799 TI - [Application the sternocleidomastoid muscle-great auricular nerve flap in radical parotidectomy]. AB - OBJECTIVE: To study a new method for repair of facial depression and facial nerve defect after parotid carcinoma resection. METHODS: 12 patients with parotid carcinoma and peripheral bone invasion were treated using facial nerve canal dissection and radical resection of the tumor, the parotid gland and the involved facial nerve and bone, including the mastoid, stylomastoid foramen, styloid process and the rear part of the mandible. A sternocleidomastoid muscle flap was elevated and transferred to repair the facial depression. The great annular nerve in the flap was anastomosed with the severed end of the facial nerve in the canal. RESULTS: The depressed deformity of the parotid area was well corrected in 9 patients. The aesthetic results were compromised in 2 patients because of tumor recurrence and reoperation. The depressed deformity was not corrected in 1 patient because of infection. Postoperatively, the function of the facial nerve recovered to a normal level. The recovery time ranged from 12 to 20 weeks ,with an average of 16.3 weeks. The local control rate of tumor was improved. CONCLUSIONS: Immediate transplantation of the sternocleidomastoid muscle-great auricular nerve flap and facial nerve canal dissection in radical parotidectomy can repair the depressed deformity of the parotid area, restore facial nerve function,and decrease tumor recurrence. The method is an ideal operation with functional recovery. PMID- 15835800 TI - [The reverse forearm flap pedicled with the ulnar branch of the ulnar artery]. AB - OBJECTIVE: To introduce a new flap for the repair of soft tissue defects in the hand. METHODS: The distally pedicled ulnar flap is based on the ascending branch of the upper-wrist cutaneous branch of the ulnar artery. The retrograde flow is ensured by the dorsal carpal arch via the descending branch. RESULTS: From 2000 to 2003, the flap was used to treat 8 cases of hand defects, including 3 dorsal defects, 2 palmar defects and 3 the first web defects. All flaps survived and the appearance was satisfactory. CONCLUSIONS: The reverse forearm flap pedicled with the ulnar branch of the ulnar artery provides a long vascular pedicle. The donor site scar is aesthetically acceptable. PMID- 15835801 TI - [Emergency repair of finger soft tissue loss with island flaps]. AB - OBJECTIVE: To introduce the applications of a series of digital island flaps to the repair of finger and/or fingertip defects and explore the indications. METHODS: A total of 79 cases (82 digits) were treated with different flaps, including (1) the digital neurovascular pedicled V-Y advancement flap; (2) the reversed dorsoulnar arterial flap of the thumb; (3) the reversed digital arterial dorsal branch flap; (4) the modified Moberg flap; (5) the reversed digital arterial flap; (6) the Hueston flap; (7) the homodigital adipofascial turnover flap and the twin neurovascular island flaps. RESULTS: Satisfactory results were obtained in digital appearance and function. CONCLUSIONS: These hoinodigital island flaps were particularly helpful for repair of finger and/or fingertip defects. PMID- 15835802 TI - [Effects of hVEGF cDNA on random skin flap via a replication-deficient adenovirus vector]. AB - OBJECTIVE: The purpose of this study was to determine the effects of local delivery of vascular endothelial growth factor( VEGF) transferred with adenovirus mediated gene on the survival of ischemic random skin flap in rats. METHODS: The animals were divided into three groups randomly (n = 10) . A 2 cm x 8 cm dorsal skin flap was designed with the pedicle at the level of the iliac crest. In group A (AdCMV-VEGF), each animal received 10(12) PR replication-deficient recombinant adenovirus (AdCMV-VEGF) in the distal two-thirds of the proposed flap by means of the subdermal injection at ten different locations. In group B (AdCMV-GaI), each received 1012 PR AdCMV-Gal. In Group C (Saline), each received 1 ml saline. Three days after the treatment, the flap was elevated as planed way and re-sutured back to its donor site. All the animals were evaluated 7 days after the operation. RESULTS: The mean percentage of surviving flap area was (85.91 +/- 2.54)% in group A, (59.56 +/- l.18)% in group B, and (61.48 +/- l.09)% in group C. There was a significant increase in the percentage of the survival area in the flaps of the group A, compared with the group B and group C (Group B vs. Group A, P < 0.01; Group C vs.Group A, P < 0.01, Group B vs. Group C, P >0.05). Hybridization in the situ, the immunohistochemical stain showed that the VEGF was expressed in the survival tissue of the flap treated with the AdCMV-VEGF, but it was not found in the control groups. Histological analysis demonstrated qualitatively greater amount of granulation tissue and angiogenesis was found in the group treated with the AdCMV-VEGF than the controls. CONCLUSIONS: The results may indicate that Ad vector carrying VEGF cDNA could be useful in enhancing the survival of the skin flap due to the effect of the local delivery of the VEGF. PMID- 15835803 TI - [Study of osteoblasts transfected with gfp in vitro and traced in vivo]. AB - OBJECTIVE: To observe the osteoblasts transfected with green fluorescent protein (GFP)by adenovirus vector expressed in vitro and traced it in vivo in order to research the feasibility of GFP as a tracer of seeding cells for tissue engineering. METHODS: CFP were transfected into the osteoblasts which derived from adult human bone marrow stromal cell (hBMSc) by adenovirus vector after being packed in 293A cells. The nontransfected hBMSc was used as the control group. The osteoblasts in each group were observed under an inverted phase contrast microscope and fluorescence microscope. The expressive efficiency of GFP was examined by flow cytometry,and alkaline phosphatase (ALP) activities and osteocalcin (OCN) synthesis. After eight days of the transfection,the osteoblasts were implanted into the muscle of nude mouse thigh while the non-transfected osteoblasts were also implanted as a control. Four and eight weeks after the operation, the nude rats were killed and the continuous tissue sections were examined using fluorescence microscopy after adjacent sections were performed by immunohistochemistry or routine HE staining. RESULTS: The green fluorescence was shown the transfected osteoblasts which derived from bone marrow. The rate of positive expression was over 75%. After eight days of the transfection, the marker proteins of the surface of the osteoblasts showed extremely efficient expression of CD29 and CD44, but the CD34 expressed negative. Either ALP or OCN of the osteoblasts was no significant difference between the two groups (P > 0.05) four and eight days after the transfection. The GFP were obviously expressed in nude mouse both at four and eight weeks, meanwhile it did not harm on the morphology and function of the transfected osteoblasts whose immunohistochemistry examination showed positive reaction. CONCLUSIONS: GFP could be transfected osteoblasts effectively in vitro and traced in vivo in nude mouse. It may be an optimal tracer for living cells on tissue engineering research. PMID- 15835804 TI - [Experimental study of human skin fibroblasts cultured in three-dimension(3D)]. AB - OBJECTIVE: To investigate the biological characters of human skin fibroblasts in fibroblast populated collagen lattice (FPCL). METHODS: The human fibroblasts were cultured in 3D and the collagen of the rat tail was also prepared. They were examined with the comprising cell cycle and apoptosis, mRNA expression of TGF beta1, and fibronectin, and cell morphology. RESULTS: The flow cytometry showed that the G0/G1, stage cells were 79% +/- 3%, 87% +/- 2% after the 7 days and 14 days separately, and there were not apoptosis peak observed. RT-PCR analysis revealed that the mRNA expression of TGF beta1, and fibronectin had no difference between human skin fibroblasts cultured in 3D and 2D. Electron microscope showed the cells were plenty of chromatin and organelles. CONCLUSIONS: The proliferation of the human skin fibroblasts in FPCL is slow, but its biological viability is better. PMID- 15835805 TI - [Effect of expression of platelet-derived growth factor B gene on blood vessel reconstruction after tissue engineering skin grafting]. AB - OBJECTIVE: To study the effect of PDGF on dermal blood vessel reconstruction by transplanted tissue-engineering skin containing PDGF-B gene to rats. METHODS: The recombined eukaryotic expression vector, pcDNA3.1-hPDGF-B, was constructed and transfected into fibroblasts mediated by LipofectAMINE. Keratinocytes + acellular dermal matrix (group A), keratinocytes + acellular dermal matrix + fibroblasts (group B), keratinocytes + acellular dermal matrix + fibroblasts with PDGF gene (group C) were recombined respectively, then transplanted them to rat dorsum and evaluated the reconstruction of blood vessels in the dermis after 2, 4, 6 week postoperation. RESULTS: In 2-4 weeks after skin grafting the vascularization rate in group C was higher than that of group B and group A. The vascularization rates in all groups had no significant differences in six weeks (P > 0.05). CONCLUSION: PDGF-B gene plays an important role in reconstruction of blood vessels in the dermis at early tissue-engineering skin grafting, which ensures the take of grafted tissue-engineering skin. PMID- 15835806 TI - [Experimental study on biological character changes of human epidermal cells during proliferation culture in vitro]. AB - [Abstract] OBJECTIVE: The study was (1) to investigate the biological character changes of human epidermal cells during proliferation culture in vitro and (2) to provide data for construction of engineered skin. METHODS: The foreskin was collected from 20 healthy children. The epidermal cells were isolated with digestion of the foreskin and cultured in vitro. Growth curve was obtained from the data of cell counting. Cell growth kinetics was observed. Meanwhile, clonal analysis and cell size measurement was performed. The rate of keratin 19 (K19) and involucrin expression-positive cells was counted by flow cytometer. Expression of K19 and involucrin mRNA was detected by RT-PCR. RESULTS: When passaged to P6, the keratinocytes from child foreskin could be expanded by (700 +/- 37) times. Flow cytometer results showed that the percentage of K19 expression-positive cells decreased from (66.97 +/- 3.14)% to (4.65 +/- 1.38)% while the percentage of involucrin expression-positive cells increased from (11.65 +/- 1.62)% to (97.03 +/- 2.66)% at P0 and P6 respectively. RT-PCR results showed that expression of K19 mRNA decreased from P0 to P6 while involucrin mRNA kept stable with passage in vitro. CONCLUSIONS: Human epidermal cells of passage 5 maintain proliferation phenotype, which are suitable for skin tissue engineering. Decrease of proliferation phenotype content is partially responsible for the proliferation capacity loss of in vitro cultured epidermal cells. PMID- 15835807 TI - [Effects of connective tissue growth factor antisense oligonucleotides on the cultured human keloid fibroblasts in vitro]. AB - OBJECTIVE: To explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid. METHODS: CTGF antisense oligonucleotides (ASODN) conjugated with isothiocyanate fluorescence was encapsulated by liposome, and then added into the human keloid fibroblast (HKF) culturing media. The intracellular distribution of CTGF ASODN was observed with fluorescence microscopy in the fixed HKF. The proliferation of HKF was measured by MIT test. The apoptosis of HKF was measured with a flow cytometer. The collagen synthesis of HKF was measured by using H3-proline incorporation method. RESULTS: The CTGF ASODN inhibited the proliferation and collagen synthesis of the HKF, compared with the control, but it increased the apoptosis after the transfection (P < 0.01). CONCLUSION: CTGF ASODN may has anti-fibrotic effects on human keloid in vitro, and the CTGF may play an important role in promoting the fibrosis of human keloid. PMID- 15835808 TI - [Maxilla reconstruction with the free iliac osteomuscular flap and simultaneous osseointegrated implant embeding]. AB - OBJECTIVE: To evaluate the clinical outcome of reconstruction of maxillary defects with vascularized iliac crest flap and simultaneous osseointegrated implant embedding. METHODS: During September to October 2003, two patients with maxillary defects from tumor resection underwent microsurgical reconstruction. The free iliac osteomuscular flap transferring and simultaneous osseointegrated implant embedding were performed to repair the defects. Three months after the reconstructive surgery, an abutment operation was preformed and denture was applied in both cases. RESULTS: The flaps survived well. Postoperative follow-up for 8 to 9 months showed that the patients obtained good zygomaxillary appearance, normal occlusion, and satisfactory pronunciation, without oronasal fistula or other serious complications. CONCLUSIONS: The free iliac crest osteomuscular flap with simultaneous osseointegrated implant embedding is an ideal, effective and cosmetically acceptable method for maxilla reconstruction. PMID- 15835809 TI - [Experimental study on inhibition of skin graft contraction with papaverine cream]. AB - OBJECTIVE: To evaluate the effect of exteral use of papaverine cream on enhancement of skin graft. METHODS: Five mini-pigs were used for the study. 20 full-thickness wounds were created on the back of each animal. Along the midline of the abdomen, a 40 cm x 2 cm full-thickness skin graft was harvested and the wound sutured.The full-thickness graft was prepared and trimmed to 2 cm x 2 cm of 0.6 mm thickness split-skin grafts, which were transplanted to each wound with tie-over bolsters. The sutures were removed 2 weeks after the operation. Twelve pairs of 100%-survived skin grafts were selected and divided into group A (the left side) and group B (the right side), with each pair locating on the same and opposite position. From the day of suture removal, 2% papaverine cream was used to group A while plain cream was used to group B. The grafts were measured and observed for healing condition at the time of suture removal and the first, second, third, fourth, fifth, and sixth month. The surface area of the graft was measured and the shrinking ratio was calculated. After the animals were killed at the sixth month, samples of the skin grafts were taken and processed with 10% formalin, routine paraffin wax and Hematoxylin-eosin staining. The tissue structure was observed and the results were analyzed statistically. RESULTS: The grafts in two groups did not have significant differences at the time of suture removal. Observations from the first to the sixth month showed that the two groups had significant differences in skin graft contracture and histological changes. HE stains demonstrated that the skin grafts in group A had less fibroblasts, more microvessels and orderly-arranged collagenous fibers. CONCLUSIONS: External use of papaverine cream can inhabit the contraction of skin grafts. It is a safe, effective, simple, and reliable method,which has the advantages of easy delivery,cheapness, less injury and infection,and no side effects. PMID- 15835810 TI - [Expression of alpha-melanocyte-stimulating hormone in human split-thickness skin graft]. AB - OBJECTIVE: To investigate the expression of alpha-melanocyte-stimulating hormone (alpha-MSH) in human split-thickness skin autograft and the role of alpha-MSH in hyperpigmented process of the grafted skin. METHODS: Immunohistochemical technique was used to detect the expression and distribution of alpha-MSH in the split-thickness grafted skin and normal skin separately. RESULTS: The expression of alpha-MSH in most of the split-thickness grafted skin was much stronger than the control skin. The positive ratio of alpha-MSH expression was 61.1% in the split-thickness grafted skin, 11.1% in the normal skin of the donor area and 16.7% in the normal skin around the recipient area. The expression of alpha-MSH in the split-thickness grafted skin was significant high, compared with the normal skins (P < 0.01). The expression of alpha-MSH in the normal skin of the donor area was no statistic remarkably differences compared to the normal skin around the recipient area. CONCLUSION: The results indicate that the expression of alpha-MSH may markedly increase in the split-thickness grafted skin and correlate with its pigmentation after the skin graft. Overexpression of alpha-MSH may play an important role in hyperpigmented process of the skin graft. PMID- 15835811 TI - [Strengthening development of pediatric otolaryngology]. PMID- 15835812 TI - [Diagnosis and following-up and analysis of pathogenesis for congenital hearing loss]. AB - OBJECTIVE: To investigate the incidence and the relevant high-risk factors of congenital hearing loss for infants from well-baby nursery and newborn intensive care unit. To explore the scientific and feasible mode of early diagnosis, following-up and early intervention. METHODS: Two-stage universal newborn hearing screening ( UNHS ) were performed by using transient otoacoustic emission ( TEOAE ). Auditory brainstem responses ( ABRs) and 40 Hz auditory event related potentials (40 Hz-AERPs) was used as diagnostic tests for those failed re screening at three-month old. Two kinds of infants who had abnormal ABRs or high risk factors of hearing loss received following-up and routine audiological evaluation from 6 months to 3 years of age. Each infant with congenital hearing loss received personalized intervention. RESULTS: The incidence of congenital hearing loss among infants who received UNHS was 5.73 per thousand. The incidence of congenital hearing loss was 20.02 per thousand in NICU and 3.67 in WBN. CONCLUSIONS: Whether passed the UNHS or not , infants from NICU should receive ABR and 40 HzAERP tests. The following-up population includes infants with abnormal ABRs or high-risk factors of hearing loss. Following-up and early intervention should be personalized. PMID- 15835813 TI - [Analyze of the clinical characteristics in children with the severe abnormal auditory brainstem response and normal distortion product otoacoustic emission]. AB - OBJECTIVE: To explore the clinic and audiological characteristics of auditory neuropathy in children. METHODS: Between 2002 and 2004, ninety-eight cases with the severe abnormal results of ABR and the normal results of DPOAE were enrolled in the study group. The mean patient age was 9 months, with a range of 1 month to 5 years. According to the evaluation of pediatric neurologist, the high-risk factors of psychomotor development retardation were statistically analyzed whether the cases were accompanied by psychomotor development retardation or not. The cases with abnormal results of ABR and DPOAE and without abnormal conductive function were selected as the cochlea lesion group. Some same age children without hearing loss were subjected as normal control group. RESULTS: The psychomotor development retardation was performed in 83.67% of cases. Hypercholesterolemia of newborn was the common high-risk factor of psychomotor development retardation (43.88% ). Severe abnormal results of ABR were occurred in 66. 07% of ears. Prolonged latency of wave I - V ABR and acoustic stapedius reflex were observed in few cases. The DPOAE amplitudes at intermediate and high frequencies were normal or slightly descending. Occurring rate of DPOAE at intermediate and high frequencies was the same as in normal control group. CONCLUSIONS: Hypercholesterolemia of newborn is the most common cause of auditory neuropathy. The auditory test results are various in children with auditory neuropathy. This character is different from it in adults. PMID- 15835814 TI - [Diagnosis and treatment of obstructive sleep apnea hypopnea syndrome in children]. AB - OBJECTIVE: To discuss the methods of diagnosis and treatment of obstructive sleep apnea hypopnea syndrome (OSAHS) in children. METHODS: 285 patients diagnosed OSAHS by X-ray for lateral rhinopharyngeal view, fibrorhinopharyngescopy and overnight polysomnography ( PSG) were reviewed retrospectively the clinical symptoms and signs. The patients whose apnea hypopnea index(AHI) > or = 5/h, blood oxygen's saturation reducing over 0.03 are diagnosed as OSAHS. 255/285 cases were performed adenoidectomy and/or tonsillectomy and 9 cases were treated with continuous positive airway pressure (CPAP). RESULTS: The major symptoms of patients with OSAHS are snoring in sleep, breathing opening mouth, apnea, hearing loss. 281/285 cases were diagnosed as adenoid and (or) tonsil hypotrophy. Two hundred- fifty five cases received surgery, adenoidectomy and tonsillectomy in 205, only adenoidectomy in 47, simple tonsillectomy in 3. The clinical symptoms of 248 cases improved evidently after operations. Improved rate is 97.2%. In 1-3 months after surgery,PSG were checked in 105 patients again: AHI, the percentage of the sleep time for saturation monitored by pulse oximetry < 0.90, the longest time of apnea are improved than pre-operation. 9 cases using CPAP received good effects, 7 of 9 cases used CPAP were in pre- and /or post- surgery. Two cases were obesity hypopnea syndrome. CONCLUSIONS: Children with OSAHS has signification them-self; PSG is the major means for diagnosing. Surgery is main method for treating; CPAP can be as the method of treating for serious OSAHS in preoperative preparation and postoperative management. PMID- 15835815 TI - [How to reduce the incidence of complication and mortality in pediatric tracheobronchial foreign body patients]. AB - OBJECTIVE: To analyze the experience for the diagnosis and treatment of pediatric tracheobronchial foreign body cases, so to the incidence of complications and mortality. METHODS: 7260 tracheobronchial foreign body cases, from 1987 through 2003, were enrolled in this study. Early diagnosis, preoperative evaluation and perioperative treatment were strongly emphasized in this group of patients. RESULTS: 7618 patients received rigid bronchoscopy and 7260 cases were confirmed with tracheobronchial foreign bodies. The confirmed diagnosis rate is 95.3%. Among the 7260 cases, 7007 (96. 4% ) patients were diagnosed and received bronchoscopic foreign body removal within 24 hours. Different food is the most common foreign body aspirated in infants and toddlers (under 4 year old), whereas older children are more likely to aspirate other foreign bodies. Peanuts and sunflower seed (6862) were noted as the most common ones. Pen caps (398) are much less. All patients were evaluated before surgery after 1996. Complications occurred in 7 patients (0.1%) . The mortality rate is zero. 7165 cases' foreign body were successfully removed the first time of brochoscopy. 95 patients received more than one times bronchoscopy. 67 cases' foreign bodies were smoothly removed with second bronchoscopy. 16 cases received flexible bronchoscopic foreign body removal when rigid bronchoscopic procedure failed. 12 cases received chest surgery for foreign body removal. 6933 cases received bronchoscopy with surface anesthesia or without anesthesia, while 327 patients with general anesthesia combined local surface anesthesia. CONCLUSIONS: Rapid diagnosis and treatment can reduce the complication and mortality rate. So that early diagnosis, preoperative evaluation and perioperative treatment should be strongly emphasized. PMID- 15835816 TI - [Apoptosis in the immune response of inner ear]. AB - OBJECTIVE: To investigate whether apoptosis is one of the mechanism in the immune response of inner ear,and to detect the expression of Fas, FasL, Bcl-2 and Bax in the inner ears. METHODS: Sixteen healthy, female guinea pigs were employed in the experiment. Sensitized systematically with keyhole limpet hemocyanin (KLH), the KLH-immunized animals were inoculated with the same antigen, and the control animals were injected PBS through cochlea basal turn. The animals were sacrificed at 7 day after inner ear vaccination. Transmission electron microscopy was used to detect inner ear apoptotic cells, and paraffin sections of cochlea from animals were stained using a terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay to identify inner ear cells undergoing apoptosis. Immunohistochemistry method was used to detect the expression of Fas, FasL, Bcl-2 and Bax in the inner ears. RESULTS: The observation of electron microscopy had shown the features characteristic of apoptotic cells in the KLH-immunized inner ears but not in the control inner ears. TUNEL-positive cells were found in the KLH-immunized inner ears but not in the control inner ears. The positive cells were the hair cells in Corti's organ, and the marginal cells in the stria vascularis and the neurons in the spiral ganglion. Moreover under morphological analysis by light microscope, these cells had the features characteristic of apoptosis. High expression of Fas and FasL could be detected in Corti's organ, the stria vascularis, the spiral ligament and the neurons of the spiral ganglion in the KLH-immunized inner ears. A low expression of Fas could be detected in the stria vascularis and the neurons of the spiral ganglion in the control inner ears, but no cells staining positive for FasL were found in the control inner ears. No cells staining positive for Bcl-2 were found in the KLH-immunized animals but moderate expression of Bcl-2 could be detected in Corti's organ, the lateral wall and the neurons of the spiral ganglion in the control inner ears. High expression of Bax could be detected in Corti's organ, the lateral wall and the neurons of the spiral ganglion in the KLH-immunized inner ears. A low expression of Bax could be detected in the neurons of the spiral ganglion and no cells staining positive for Bax were found in Corti's organ, the lateral wall in the control inner ears. CONCLUSIONS: These findings suggest apoptosis is involved in the pathogenesis of the immune response of inner ear and Fas- FasL pathway is one of important signal transportation of the course and Bcl-2 and Bax have a critical role in the regulation of apoptotic cell death induced by the immune response of inner ear. PMID- 15835817 TI - [Roles of epidermal growth factor and its receptor on the chronic tympanic membrane perforations]. AB - OBJECTIVE: To evaluate the possible roles of epidermal growth factor(EGF) and its receptor (EGFR) on the chronic tympanic membrane perforations. METHODS: A phosphate buffer saline of EGFR was administered to a Gelfoam pledget placed over chronic tympanic membrane perforations in guinea pigs. The EGFR of 10 specimens from the acquired middle ear cholesteatoma of the adjacent skin around perforation was examined by immunohistochemical SP method and computer image analysis. Results Complete closure of the tympanic membrane perforations was observed in 82.6% of EGF-treated ears, but only 33.3% in the controls(P < 0. 01). No case was led to middle ear cholesteatoma in the experiment group (0/23). The positive expression in the adjacent skin around perforation was (39.3 -/+ 7.4)%; and the normal external ear skin was (25.4 +/- 3.7)%; There were distinctly significant differences between the adjacent skin around perforation and the normal external ear skin (P < 0. 01). CONCLUSIONS: EGF is effective on closing chronic tympanic membrane perforations in the guinea pigs. Present data suggests that EGF-treated may induce the occurrence of middle ear cholesteatoma. PMID- 15835818 TI - [Effect of intranasal glucosteroid on the gene expression of interleukin-5 in nasal polyps]. AB - OBJECTIVE: To investigate the effect of intranasal glucosteroid treatment on the expression of interleukin (IL)-5 gene in nasal polyps. METHODS: Nasal polyps from topical steroid treated patients (n = 20) and untreated patients (n = 20) were investigated with the technique of mRNA in situ hybridization. RESULTS: The majority of IL-5 mRNA positive cells in nasal polyps were lymphocytes or eosinophils. No statistical significance was found in the densities of IL-5 mRNA positive cells between allergic patients [(12.6 +/- 4.6)/0.25mm2] and nonallergic patients [(14.3 +/- 4.1)/0.25mm2] (t = -0.775, P > 0.05). Compared with the control group [(13.9 +/- 4.2)/0.25mm2], the density of IL-5 mRNA positive cells was decreased in the steroid-treated group [(10. 2 +/- 3.1)/0.25mm2], and the difference reached statistical significance (t = 3.114, P < 0.01). CONCLUSIONS: These findings indicate that topical steroid treatment may suppress the IL-5 gene expression, and steroids may inhibit eosinophil functions. PMID- 15835819 TI - [Nasal alar reconstruction with a vascularized free auricular composite flap]. AB - OBJECTIVE: The plastic surgeons prefer to reconstruct nasal alar with free auricular composite flap because it well matches nasal tissue in contour, texture and color. However, the size of the free composite flap should be less than 1. 0 cm x 1.5 cm due to the limitation of revascularization. Our aim is to search for a surgical method which could be used to repair full-layer larger nasal alar defects. METHODS: A surgical technique was presented to repair nasal alar defect with a free auricular composite flap, which was vascularized by branches of superficial temporal artery. Briefly, the contralateral auricular composite tissue pedicled by superficial temporal vessels (3 to 4 cm in length) was harvested from region of helix crus and preauricular skin, which matched the arc of the nasal rim, and then transplanted onto the recipient area. The superficial temporal vessel pedicles were anastomosed to facial vessels via vessel grafts harvested from lateral circumflex femoral vessels, which were about 10 to 12 cm in length. RESULTS: Twelve cases were treated, among them, 11 cases were successfully repaired with satisfactory results, one case failed, possibly due to the bad condition of the patient's blood vessels. In our cases, the size of alar defect varied from 1.5 cm x 2.5 cm to 2.0 cm x 3.8 cm. The minimal auricular donor site deformities and inconspicuous scar were acceptable and could easily be hidden by hair. CONCLUSIONS: The technique of free auricular composite flap, which is vascularized by superficial temporal vessels, is ideal for nasal alar reconstruction. PMID- 15835820 TI - [Effects of basic fibroblast growth factor on protein kinase B activity and c-fos expression in CNE- I nasopharyngeal carcinoma cell line]. AB - OBJECTIVE: The relationship between basic fibroblast growth factor (bFGF) and phosphatidylinositol 3-kinase/protein kinase B(PI 3-K/PKB) signal pathway was studied during the course of bFGF regulating carcinoma epithelioid cell lines- I (CNE- I ) nasopharyngeal carcinoma cell proliferation through altering time of bFGF treatment. METHODS: PKB activities were measured by advanced Takai method. C fos expression was examined by Western Blot. RESULTS: bFGF (50 ng/ml) significantly stimulated PKB activity,After pretreatment of Wortmannin for 1 h, the PKB activity decreases to the basal levels(P < 0. 05),compared with control group. After treatment of bFGF (50 ng/ml) at various time, the expression of c fos is increased. The expression of c-fos increases to the peak when the time of bFGF treatment is 30 min. After pretreatment of Wortmannin for 1 h, the expression of c-fos decreases 20% when the time of bFGF treatment was 30 min and when the time of bFGF treatment was 45 min, the expression of c-fos decreases 35%. CONCLUSIONS: 1. PI 3-K and PKB mediate bFGF-induced signal transduction in CNE- I nasopharyngeal carcinoma cell line; PKB lies in downstream of PI 3-K. 2. bFGF induces the increased expression of c-fos through PI 3-K/PKB in CNE- I nasopharyngeal carcinoma cell line. PMID- 15835821 TI - [Research of fibronectin, p53 and Ezrin protein with occurrence, progress and prognosis in the primary laryngeal carcinoma by the tissuechip technology]. AB - OBJECTIVE: To evaluate the predictive role of fibronectin, p81 (Ezrin protein) and p53 gene in primary laryngeal carcinoma, it's relationship with epidemiology(smoking), histological grading, surgical treatment, TNM stage and prognosis were studied by the tissuechip technology. METHODS: The expression of fibronectin, p53 gene and p81 (Ezrin protein) on a series of 85 primary laryngeal carcinoma patients treated in our hospital between 1992 and 2000 was studied with tissuechip technology. The correlation of each score according to the intensity and percentage of labeled cells or intercellular substance with relevant clinical dada was statistically analyzed. RESULTS: Some cases were lost or boosted no tumor tissue in our tissuechip. Among the 70 cases available, 45.71% (32/70) of the specimens' basal membrane and extracellular matrix were strongly stained with fibronectin; there is statistical significance (P < 0.05) between primary tumor grading groups. Ezrin protein expressing rate is 87.3%, and the average percentage of its labeled cells is 53.68% (ranging from 0% to 100%, median is 58. 69%). There were significant difference between tumor grading groups, clinical early and late stages and 3-year survival rates (P < 0.05) after chi-square test. But no relation with smoking, gender, age and histological classes (P > 0.05). The average percentage of p53 positive cells is 21.6% (ranging from 0% to 90.3%, median is 5.85%) and 46.8% showed positive stains in our research. There was no statistical prominence in p53 protein demonstration between TNM stages, lymph node metastasis, 3-year survival rate, smoking, gender, age and histological classes (P > 0.05). CONCLUSIONS: The tissue microarray technique spent shorter time and less expense, and showed higher consistency in our essays. And the present study suggests fibronectin and p81 (Ezrin protein) could be the clinical discriminators in laryngeal carcinoma. PMID- 15835822 TI - [Culture of neural stem cells from rat olfactory bulb]. AB - OBJECTIVE: To establish the method of in vitro culture of neural stem cells (NSC) from rat olfactory bulb and investigate the characteristics of its proliferation and differentiation. METHODS: NSC from postnatal one-day (P1) and adult rat olfactory bulb were isolated and cultured in serum free media with epidermal growth factor (EGF) and fibroblast growth factor-basic (bFGF). Antibodies against NSC (nestin), neuronal (neuronal specific enolase, NSE), astrocytic (glial fibrillary acidic protein, GFAP) and oligodendrocytic (galactocerebroside, GalC) markers were used to identify NSC and specific neural cells differentiated from NSC with immunocytochemical staining. Growth curve of olfactory bulb NSC was measured using MTT method. RESULTS: Nestin immuno-positive NSC were isolated and cultured from P1 and adult rat olfactory bulb which could differentiate into neurons, astrocytes and oligodendrocytes. The forming rates of neurosphere from P1 and adult rat olfactory bulb were 20% approximately 30% and 0.1% respectively. The proliferation of olfactory bulb NSC depended on EGF and bFGF, in which EGF increased proliferation of cells stronger than bFGF. CONCLUSIONS: NSC with self renewal capacity and potential multi-differentiation were cultured from P1 and adult rat olfactory bulb. PMID- 15835823 TI - The Americans with Disabilities Act of 1990 and its impact on higher education and nursing education. AB - This paper outlines the Americans with Disabilities Act of 1990 (ADA) and explores the extensive impact the ADA legislation has had on institutions that provide higher education and nursing education in the United States. PMID- 15835824 TI - Changes in steps per day over the course of a pilot walking intervention. AB - The objective of this study is to explicate changes in steps per day observed over the course of a pilot study concerned with promoting walking. A pre/post single group design was used to evaluate a 12-month group intervention with 6 month follow-up with a sample of twenty-one sedentary, hypertensive African American women 25-68 years of age. A 3-hour monthly meeting was paired with an at home walking component. Results indicated that the total group experienced a slight increase (5%) in mean steps per day at the end of the 12-month intervention, while a subgroup experienced a 13% decrease. However, mean steps per day increased from baseline to 6-month follow-up for both groups. Mean steps per day increased from 3857 to 5582 (37%) for total group and from 4659 to 7038 (51%) for the subgroup. CONCLUSIONS: A group intervention can have a positive impact on promoting walking among African American women; however, multi-phased interventions may be necessary to assist this population meet current physical activity recommendations. PMID- 15835825 TI - The impact of calculated ambulation on health promotion. AB - A study to assess the impact of calculated ambulation on health promotion was conducted over a five-month period at a southern university. The study, which initially included approximately forty-three healthcare management students, was based on a similar study conducted at a larger university in South Carolina. The study at the larger university showed a connection between the number of steps one took and one's health/fitness status. With a 10,000 steps a day intervention /walking plan, pre- and post-tests on weight, blood pressure and cholesterol levels were conducted with the assistance of the local County Health Department. Individualized longitudinal case profiles were then presented, with an overview of program benefits; the role of support persons and the examination of the individual responsibility for health concept. PMID- 15835826 TI - A qualitative perspective of barriers to health-promoting behaviors of African Americans. AB - African American mortality and morbidity rates remain alarmingly high. Implementation of health promotion strategies can be effective in reducing these rates. Yet, African American health-promoting behaviors remain inadequate. Exploration of perceived barriers to implementing health-promoting behaviors from a qualitative perspective may lead to a better understanding of African American barriers to healthier lifestyles. The purpose of this study was to explore barriers to health promotion for African Americans from a qualitative perspective. Focus group interviews were held in two southeastern states. The results yielded three themes. These were cost, lack of discipline versus not having enough time, and a lack of motivation. When health care professionals develop treatment strategies for African Americans, these barriers should be considered to aid in the development of more efficacious plans of care. PMID- 15835827 TI - Learning styles: an issue in clinical education? PMID- 15835828 TI - Fire in the operating room during tracheotomy: a case report. AB - We describe a patient who sustained facial burns during a tracheotomy. The electrosurgical unit indirectly started the fire during monitored anesthesia care when a high inspired oxygen concentration was being delivered to the patient by simple face mask. This case points out the need for prevention strategies, intraoperative vigilance, and quick intervention to prevent further patient injury any time the electrosurgical unit is used in an oxygen-enriched atmosphere. PMID- 15835829 TI - Anesthetic considerations for patients with Down syndrome. AB - Patients with Down syndrome, or trisomy 21, present a unique set of anesthetic considerations to anesthesia providers. Down syndrome is the most prevalent genetic disorder worldwide and affects more than 1 in 800 live births. Patients with Down syndrome are afflicted with multiple congenital anomalies that affect almost all of their organ systems. Skillful management during the perioperative period is essential for a good outcome for patients with multiple congenital abnormalities in the cardiopulmonary and musculoskeletal systems. The purpose of this review is to describe clinically significant findings that are common to patients with Down syndrome. In addition to the presentation of clinical anomalies associated with trisomy 21, specific anesthetic considerations and interventions are reviewed. PMID- 15835830 TI - Case study involving suctioning of an electromyographic endotracheal tube. AB - The electromyographic endotracheal tube (EMG-ETT) is a relatively new tool used to measure integrity of the vocal cord structures during surgery. We describe a case in which an EMG-ETT was inserted for the operative period but not replaced with an ETT during the immediate postoperative period. Intensive care unit nurses had difficulty suctioning the EMG-ETT. The patient was not provided the pulmonary toilet necessary until the EMG-ETT was removed and replaced with a regular ETT. The purpose of this article is to make anesthesia providers aware that when mechanical ventilation is required during the postoperative period, the EMG-ETT should be removed and replaced with a regular ETT to facilitate pulmonary toilet. PMID- 15835831 TI - An anesthesia provider's perspective of heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a pathology manifested as clinically induced destruction of platelets. There are 2 forms of HIT, type I and type II; type II is the more serious. HIT type I is a transient, non-immune-mediated form manifesting with mild thrombocytopenia. Type II is a drug-induced, immune mediated syndrome that may cause life- or limb-threatening thromboembolic events. Induction of general anesthesia for a person with HIT type II is no different from that for a person in the general population. Treatment modalities vary only if heparin will be used during the case. The initial indicator of HIT is decreased platelet count, with or without thrombosis. Clinical criteria and advanced serological testing are available for the definitive diagnosis of HIT. Clinical suspicion of HIT remains key to early cessation of heparin (all routes) and initiation of alternative treatments. PMID- 15835833 TI - AANA Journal Course: update for nurse anesthetists. Arrhythmia management devices and electromagnetic interference. AB - The technological complexity of implantable arrhythmia management devices, specifically pacemakers and defibrillators, has increased dramatically since their introduction only a few decades ago. Patients with such devices are encountered much more frequently in hospitals and surgery centers, yet anesthesia provider knowledge of safe and proper management is often incomplete. Anesthesia textbooks and references may provide only short paragraphs on arrhythmia management devices that do not address important perioperative management strategies for this ever-growing patient population. It is no longer satisfactory to simply place a magnet over an implanted device during surgery and assume that this action protects the patient from harm due to electromagnetic interference from inappropriate device function. This AANA Journal course serves as a concise review of basic device function, the sources and effects of electromagnetic interference in the operative setting, and patient management recommendations from current literature. PMID- 15835834 TI - How have your protocols changed over your years of practice? Is it easier now or when you began midwifery? PMID- 15835832 TI - The effect of Pencan needle orientation on spinal anesthesia outcomes. AB - Slow resolution of block and incidence of side effects deter many practitioners from choosing spinal anesthesia for out-patient surgical procedures. Some studies suggest that controlling bevel or side port orientation of a spinal needle during anesthetic injection can affect occurrence of side effects and time to block resolution. The objective of this study was to determine the effects of varying Pencan spinal needle (B-Braun, Bethlehem, Pa) side port orientations on duration of block and incidence of side effects in groups of patients receiving spinal anesthesia. We randomized 87 subjects scheduled for spinal anesthesia to receive a spinal anesthesia injection using a cephalad, lateral, or caudad side port orientation. Onset, duration, block height, incidence of side effects, and analgesic requirements were among the variables measured. No difference in onset, duration, or analgesic requirements was noted among groups. Differences were noted in time to discharge from the hospital (P = .027) and time to first voiding (P = .023) in the lateral compared with the cephalad and caudad orientation groups. Patients in whom the lateral needle side port orientation was used for injection were discharged earlier and had fewer side effects. This could translate into significant savings, financially and in terms of staff requirements. PMID- 15835835 TI - Marion's message. Unchanging protocols. PMID- 15835836 TI - Protocols: finding the balance. PMID- 15835837 TI - Can the art of midwifery survive protocols? PMID- 15835838 TI - Protocols. PMID- 15835839 TI - The challenge of doing nothing. PMID- 15835840 TI - Jasmine's story: what are the limits of "normal birth"? PMID- 15835841 TI - The air we breathe: oxygen: tradition and heresy? PMID- 15835842 TI - Problematizing choice in the elective cesarean debate. PMID- 15835843 TI - The new kid on the block: 30 years and growing. PMID- 15835844 TI - Screening tests: not worth triple the trouble. PMID- 15835845 TI - Belly dance & birth. PMID- 15835846 TI - Antiretroviral basics. PMID- 15835847 TI - Metamorphosis. PMID- 15835848 TI - The blessing of a Blessingway. PMID- 15835849 TI - Having Ahna at home. PMID- 15835850 TI - The Bristol Third-Stage Trial. PMID- 15835851 TI - The business of midwifery: contracting. PMID- 15835852 TI - Merging technology and tradition. PMID- 15835853 TI - HIV in South Africa. PMID- 15835854 TI - Doctor-midwife exchange. PMID- 15835855 TI - Observations from Hungary. PMID- 15835856 TI - Midwifery in Pakistan. PMID- 15835857 TI - Birth and death in Yayasan Bumi Sehat, Bali. PMID- 15835858 TI - A progressive midwife in Nazareth. PMID- 15835859 TI - Hari's birth. PMID- 15835860 TI - Licensure examination. PMID- 15835861 TI - Ethics. PMID- 15835862 TI - Ethics. PMID- 15835863 TI - Keeping patients happy in your waiting room. PMID- 15835864 TI - You can't afford to lose your data. PMID- 15835865 TI - Discover the advantages of claims-made coverage. PMID- 15835868 TI - Crystallized frameworks with giant pores: are there limits to the possible? AB - In the domain of porous solids with inorganic or hybrid frameworks, the combination of mastered chemistry and of computer simulations pushes forward the limits of the classical approach and allows the full determination from powder diffraction data of architectures with cells of several hundred thousand cubic angstroms with hierarchies of giant pores and unprecedented Langmuir surfaces. The different limits induced by this new approach are analyzed. PMID- 15835869 TI - Inorganic-organic hybrid molecular architectures of cyanometalate host and organic guest systems: specific behavior of the guests. AB - Molecular architectures built of inorganic cyanometalate building blocks provide variegated host structures with several organic guest molecules. The strategies to derive novel structures are presented briefly. The formation of a charge transfer (CT) complex as the guest inside the cavity and the photochemistry of the CT complexes are discussed. The chemical pressure that the guest experiences inside the cavity is also discussed based on the vibrational spectroscopic results. PMID- 15835870 TI - Fullerene-porphyrin constructs. AB - Porphyrins and fullerenes are spontaneously attracted to each other. This new supramolecular recognition element can be used to construct discrete host-guest complexes, as well as ordered arrays of interleaved porphyrins and fullerenes. The fullerene-porphyrin interaction underlies successful chromatographic separations of fullerenes, and there are promising applications in the areas of porous framework solids and photovoltaic devices. PMID- 15835871 TI - Ligand design in multimetallic architectures: six lessons learned. AB - Metallosupramolecular chemistry involves the use of combinations of organic ligands and metals for the construction of both discrete and polymeric aggregates. This Account describes some lessons that we have learned about aspects of ligand design in the course of our work in this area. Specifically, we recommend the incorporation of a diverse range of heterocyclic rings and arene cores within the ligands, as well as attention to symmetry considerations, and offer suggestions for the introduction of chirality and flexibility within the ligands and the exploitation of other weak interactions to assist self-assembly processes. PMID- 15835872 TI - Small amphiphilic organics, coordination extended solids, and constant curvature structures. AB - In this Account, we explore the role constant curvature structures play in amphiphilic small molecule crystals and extended coordination solids. A constant curvature structure is one in which there is a surface or interface that has the same curvature throughout its surface. Simple examples of such structures contain spheres (micelles), columns, or layers. Yet another family are cubic as is found in the gyroid topology. For amphiphilic systems, there are two domains, one generally hydrophobic and the other hydrophilic. We find that the interfaces between these two domains in small molecule structures resemble those in larger scale systems and adopt topologies related to constant curvature structures. The hydrophobic-to-total volume ratio, a molecular parameter, can be used to predict which type of constant curvature structure is adopted. In the case of three coordinate extended solids, constant curvature plays a role both in the extended net topology and in the hydrophobic-to-hydrophilic interface. PMID- 15835873 TI - 3-D molecular assembly of function in titania-based composite material systems. AB - Various examples of composite titania-based nanostructured materials exhibiting cooperative functionalities between different active components are presented. The fabrication of these integrated composite materials is based on one-pot supramolecular templating techniques combined with acidic sol-gel chemistry. The defined 3-D nanoscale organization and integration of various functional components results in advanced optoelectronic and photonic applications such as visible light sensitization of mesoporous titania photocatalysts with cadmium sulfide nanocrystals acting as sensitizing integral part of the mesopore wall structure, narrow bandwidth emission from rare earth ion activated nanocrystalline mesoporous titania films, and mirrorless lasing in dye-doped hybrid organic/inorganic mesostructured titania waveguides. PMID- 15835874 TI - Design, chirality, and flexibility in nanoporous molecule-based materials. AB - Scientific and technological interest in porous materials with molecule-sized channels and cavities has led to an intense search for controlled chemical routes to systems with specific properties. This Account details our work on directing the assembly of open-framework structures based on molecules and investigating how the response of nanoporous examples of such materials to guests differs from classical rigid porous systems. The stabilization of chiral nanoporosity by a hierarchy of interactions that both direct and maintain a helical open-framework structure exemplifies the approach. PMID- 15835875 TI - Microporous porphyrin solids. AB - Metalloporphyrins are exceedingly useful building blocks for the design and synthesis of molecularly based solids. The use of hydrogen bonding or metal ion coordination provides a wide range of framework solids. Using various polyfunctionalized porphyrins, we have created porous solids that are thermally robust and that retain their internal porosity upon loss of solvates. Their pore dimensions are comparable to zeolites, and they show shape and size selectivity in sorption of guest molecules and in epoxidation of alkenes. PMID- 15835876 TI - The interface chemistry between chalcogenide clusters and open framework chalcogenides. AB - One of the most exciting recent developments concerning molecular architectures is the emerging field of crystalline chalcogenide superlattices that bridges two traditional but distinct areas of research: chalcogenide clusters and porous materials. By combining synthetic and structural concepts in these two areas, many crystalline solids containing spatially organized chalcogenide clusters have been created that exhibit varied properties ranging from microporosity, fast ion conductivity, and photoluminescence to narrow and tunable electronic band gaps. The potential applications of these materials extend beyond traditional areas such as acid catalysis or adsorption-based separation to include shape- or size selective photocatalysis, solid-state ionics, and electrochemistry. PMID- 15835877 TI - Past, present, and future of periodic mesoporous organosilicas-the PMOs. AB - Periodic mesoporous organosilicas (PMOs) represent an exciting new class of organic-inorganic nanocomposites targeted for a broad range of applications such as catalysis and sensing, separations, and microelectronics. Their hallmark is the presence of organic bridging groups incorporated into the channel walls of an ordered nanoporous structure, which represents a useful tool to finely tune the chemical and physical properties of the materials. We discuss the history of the discovery and development of the PMOs emphasizing the most important recent advancements regarding compositions and structures, morphologies, and properties. Furthermore, we present an outlook about the promising future perspectives of PMOs that result from the latest developments in this field. PMID- 15835878 TI - Molecular tectonics: from simple tectons to complex molecular networks. AB - Molecular networks in the crystalline phase are infinite periodic molecular assemblies formed under self-assembly conditions between self-complementary or complementary tectons. These millimeter-size structures may be regarded as hypermolecules formed by supramolecular synthesis using reversible intertecton interactions. Molecular tectonics, based on molecular recognition events and their iteration, is the approach dealing with design and preparation of molecular networks in the solid state. In this Account, an overview of the rational behind this approach is presented. A variety of molecular networks based on van der Waals interactions and hydrogen and coordination bonding possessing diverse connectivity and topology are discussed. PMID- 15835879 TI - Directed assembly of metal-cyanide cluster magnets. AB - The simple, well-understood coordination chemistry of the cyanide ligand is of significant utility in the design of new single-molecule magnets. Its preference for bridging two transition metals in a linear M'-CN-M geometry permits the use of multidentate blocking ligands in directing the assembly of specific molecular architectures. This approach has been employed in the synthesis of numerous high nuclearity constructs, including simple cubic M4M'4(CN)12 and face-centered cubic M8M'6(CN)24 coordination clusters, as well as some unexpected cluster geometries featuring as many as 27 metal centers. The ability to substitute a range of different transition metal ions into these structures enables adjustment of their magnetic properties, facilitating creation of high-spin ground states with axial magnetic anisotropy. To date, at least four different cyano-bridged single molecule magnets have been characterized, exhibiting spin-reversal barriers as high as 25 cm(-1). Ultimately, it is envisioned that this strategy might lead to molecules possessing much larger barriers with the potential for storing information at more practical temperatures. PMID- 15835880 TI - New approaches to the analysis of high connectivity materials: design frameworks based upon 4(4)- and 6(3)-subnet tectons. AB - Coordination framework polymers derived from lanthanide metal ions with N,N' dioxide ligands (4,4'-bipyridine-N,N'-dioxide, pyrazine-N,N'-dioxide, 1,2 bis(pyridin-4-yl)ethane-N,N'-dioxide, trans-1,2-bis(pyridin-4-yl)ethene-N,N' dioxide) exhibit such intricate architectures that a new strategy is required to appreciate and understand their structures. Rather than analyzing the overall structure in terms of the connectivity of individual metal nodes, which can lead in some cases to extremely complex topological treatments, our new strategy is based on the visualization of the structures as combinations of interconnected layered 2-D sheets or subnet tectons. Despite the diversity and relative complexities of many of the structures discussed here, they can all be described by the interconnection of just two types of 2-D subnet tectons, 4(4) square grids or 6(3) hexagonal grids. The interconnection of these layered sheets with bridging N,N'-dioxide molecules gives rise to both 2-D bilayer and 3-D network extended structures depending upon the relative dispositions of the interconnecting N,N'-dioxide ligands. Thus, 2-D bilayers result when the N,N' dioxide ligands that bridge two subnet tectons are located on the same side of the sheet, while 3-D networks are formed when the bridging N,N'-dioxide ligands are located on both sides of the sheet. This analysis allows ready identification and interpretation of some of the most highly connected and complex architectures yet observed in materials chemistry. PMID- 15835881 TI - Selective molecular recognition, C-H bond activation, and catalysis in nanoscale reaction vessels. AB - Supramolecular chemistry represents a way to mimic enzyme reactivity by using specially designed container molecules. We have shown that a chiral self assembled M4L6 supramolecular tetrahedron can encapsulate a variety of cationic guests with varying degrees of stereoselectivity. Reactive iridium guests can be encapsulated, and the C-H bond activation of aldehydes occurs with the host cavity controlling the ability of substrates to interact with the metal center based upon size and shape. In addition, the host container can act as a catalyst by itself. By restricting reaction space and preorganizing the substrates into reactive conformations, it accelerates the sigmatropic rearrangement of enammonium cations. PMID- 15835882 TI - Structural evolution and phase homologies for "design" and prediction of solid state compounds. AB - The challenge of designing solid state compounds with predicted compositions and structures could be partly met using concepts that employ phase homologies. Homologous series of compounds not only can place seemingly diverse phases into a single context; they can also forecast with high probability specific new phases. A homologous series is expressed in terms of a mathematical formula that is capable of producing each member. Within a homologous series, the type of fundamental building units and the principles that define how they combine remain preserved, and only the size of these blocks varies incrementally. In this Account, we present this approach by discussing a number of new homologies and generalize it for a wide variety of systems. PMID- 15835883 TI - Coordination assemblies from a Pd(II)-cornered square complex. AB - The [enPd(II)]2+ (en = ethylenediamine) unit has emerged as a versatile building block in molecular self-assembly. In particular, the 90 degrees coordination angle of the metal has been judiciously used in the design of new discrete two- and three-dimensional structures. Our last 15 years of work with the Pd(II) cornered unit is summarized in this Account, from the spontaneous formation of a Pd4 square metal complex to a family of architectures such as cages, bowls, boxes, tubes, catenanes, and spheres. PMID- 15835884 TI - The origins of high-affinity enzyme binding to an extrahelical DNA base. AB - Base flipping is a highly conserved strategy used by enzymes to gain catalytic access to DNA bases that would otherwise be sequestered in the duplex structure. A classic example is the DNA repair enzyme uracil DNA glycosylase (UDG) which recognizes and excises unwanted uracil bases from DNA using a flipping mechanism. Previous work has suggested that enzymatic base flipping begins with dynamic breathing motions of the enzyme-bound DNA substrate, and then, only very late during the reaction trajectory do strong specific interactions with the extrahelical uracil occur. Here we report that UDG kinetically and thermodynamically prefers substrate sites where the uracil is paired with an unnatural adenine analogue that lacks any Watson-Crick hydrogen-bonding groups. The magnitude of the preference is a striking 43000-fold as compared to an adenine analogue that forms three H-bonds. Transient kinetic and fluorescence measurements suggest that preferential recognition of uracil in the context of a series of incrementally destabilized base pairs arises from two distinct effects: weak or absent hydrogen bonding, which thermodynamically assists extrusion, and, most importantly, increased flexibility of the site which facilitates DNA bending during base flipping. A coupled, stepwise reaction coordinate is implicated in which DNA bending precedes base pair rupture and flipping. PMID- 15835885 TI - Relocation of water molecules between the Schiff base and the Thr46-Asp96 region during light-driven unidirectional proton transport by bacteriorhodopsin: an FTIR study of the N intermediate. AB - A key event in light-driven proton pumping by bacteriorhodopsin is the formation of the L intermediate, whose transition to M is accompanied by the first proton transfer step, from the Schiff base to Asp85 on the extracellular side. Subsequent reprotonation of the Schiff base from the other side of the membrane to form the N intermediate is crucial for unidirectional proton transport. Previous FTIR studies have suggested that the intense water O-D stretching vibration bands which appear in L at 2589, 2605, and 2621 cm(-)(1) are due to a cluster of polarized water molecules connecting the Schiff base to the Thr46 Asp96 region closer to the cytoplasmic surface. In the present study the difference spectrum was obtained of the N intermediate with its photoproduct N', formed after irradiating N at 80 K. The water O-D stretching vibrations of N appear as a broad feature in a similar frequency region with a similar intensity to those of L. This feature is also affected by T46V like in L. However, the intensities of these water vibrations of N nearly returned to the initial unphotolyzed state upon formation of N', unlike those of L which are preserved in L'. An exception was V49A, which preserved the intense water vibrations of N in N'. The results suggest that both L and N have a water cluster extending from the Schiff base to Thr46. The surrounding protein moiety stabilizes the water cluster in L, but in N it is stabilized mostly by interaction with the Schiff base. PMID- 15835886 TI - Anion binding properties of reduced and oxidized iron-containing superoxide dismutase reveal no requirement for tyrosine 34. AB - We report the first spectroscopic observation of substrate analogue binding to the reduced state of iron superoxide dismutase from Escherichia coli (Fe(2+)SOD) and demonstrate that the pH dependence reflects inhibition of anion binding by ionized Tyr34, not loss of a positive contribution on the part of Tyr34's labile proton. This can also explain the pH dependence of the K(M) of Fe(2+)SOD. Thus, it appears that substrate binding to Fe(2+)SOD occurs in the second sphere and is not strongly coupled to hydrogen bond donation. Parallel investigations of substrate analogue binding to the oxidized state (Fe(3+)SOD) confirm formation of a six-coordinate complex and resolve the apparent conflict with earlier nuclear magnetic relaxation dispersion (NMRD) results. Thus, we propose that two F(-) ions can bind to the oxidized Fe(3+)SOD active site, either displacing the coordinated solvent or lowering its exchange rate with bulk solvent. We show that neutral Tyr34's unfavorable effect on binding of the substrate analogue N(3)(-) can be ascribed to steric interference, as it does not apply to the smaller substrate analogues F(-) and OH(-). Finally, we report the first demonstration that HS(-) can act as a substrate analogue with regard both to redox reactivity with FeSOD and to ability to coordinate to the active site Fe(3+). Indeed, it forms a novel green complex. Thus, we have begun to evaluate the relative importance of different contributions that Tyr34 may make to substrate binding, and we have identified a novel, redox active substrate analogue that offers new possibilities for elucidating the mechanism of FeSOD. PMID- 15835887 TI - Proteomic analysis of mammalian oligosaccharyltransferase reveals multiple subcomplexes that contain Sec61, TRAP, and two potential new subunits. AB - Oligosaccharyltransferase (OST) catalyzes the cotranslational transfer of high mannose sugars to nascent polypeptides during N-linked glycosylation in the rough endoplasmic reticulum lumen. Nine OST subunits have been identified in yeast. However, the composition and organization of mammalian OST remain unclear. Using two-dimensional Blue Native polyacrylamide gel electrophoresis/sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry, we now demonstrate that mammalian OST can be isolated from solubilized, actively engaged ribosomes as multiple distinct protein complexes that range in size from approximately 500 to 700 kDa. These complexes exhibit different ribosome affinities and subunit compositions. The major complex, OSTC(I), had an apparent size of approximately 500 kDa and was readily released from ribosome translocon complexes after puromycin treatment under physiological salt conditions. Two additional complexes were released only after treatment with high salt: OSTC(II) ( approximately 600 kDa) and OSTC(III) ( approximately 700 kDa). Both remained stably associated with heterotrimeric Sec61alphabetagamma, while OSTC(III) also contained the tetrameric TRAP complex. All known mammalian OST subunits (STT3-A, ribophorin I, ribophorin II, OST48, and DAD1) were present in all complexes. In addition, two previously uncharacterized proteins were also copurified with OST. Mass spectrometry identified a 17 kDa protein as DC2 which is weakly homologous to the C-terminal half of yeast Ost3p and Ost6p. The second protein (14 kDa) was tentatively identified as keratinocyte-associated protein 2 (KCP2) and has no previously known function. Our results identify two potential new subunits of mammalian OST and demonstrate a remarkable heterogeneity in OST composition that may reflect a means for controlling nascent chain glycosylation. PMID- 15835888 TI - Elucidating the substrate specificity and condensation domain activity of FkbP, the FK520 pipecolate-incorporating enzyme. AB - Rapamycin, FK506, and FK520 are potent immunosuppressant natural product macrocycles generated by hybrid polyketide synthase (PKS)/nonribosomal peptide synthetase (NRPS) systems in streptomycetes. An important functional element within these molecules is an l-pipecolate moiety that is incorporated into the completed polyketide chain by the action of RapP/FkbP, a four-domain NRPS that also putatively serves to cyclize the chain after amino acid insertion. Here we report the expression and purification of recombinant FkbP from the FK520 biosynthetic pathway. Using a combination of radioassays and Fourier transform mass spectrometry, we demonstrate that once FkbP has been phosphopantetheinylated in vitro, its peptidyl carrier protein domain can be successfully loaded with l pipecolic acid and, to a lesser extent, l-proline. The first condensation domain of FkbP is shown to be active through the successful acetylation of aminoacyl-S FkbP using the appropriately loaded terminal acyl carrier protein from the PKS array, FkbA, as the chain donor. Site-directed mutagenesis confirmed that the N terminal condensation domain catalyzes the transfer reaction. Acetylation of prolyl-S-FkbP was more rapid and occurred to a greater extent than that of pipecolyl-S-FkbP, a trend which was also observed with alternative acyl chain donors. These observations suggest that the adenylation domain of FkbP serves as the primary selectivity filter for pipecolate incorporation. PMID- 15835889 TI - Abeta40-Lactam(D23/K28) models a conformation highly favorable for nucleation of amyloid. AB - Recent solid-state NMR data (1) demonstrate that Abeta(1)(-)(40) adopts a conformation in amyloid fibrils with two in-register, parallel beta-sheets, connected by a bend structure encompassing residues D(23)VGSNKG(29), with a close contact between the side chains of Asp23 and Lys28. We hypothesized that forming this bend structure might be rate-limiting in fibril formation, as indicated by the lag period typically observed in the kinetics of Abeta(1)(-)(40) fibrillogenesis. We synthesized Abeta(1)(-)(40)-Lactam(D23/K28), a congener Abeta(1)(-)(40) peptide that contains a lactam bridge between the side chains of Asp23 and Lys28. Abeta(1)(-)(40)-Lactam(D23/K28) forms fibrils similar to those formed by Abeta(1)(-)(40). The kinetics of fibrillogenesis, however, occur without the typical lag period, and at a rate approximately 1000-fold greater than is seen with Abeta(1)(-)(40) fibrillogenesis. The strong tendency toward self-association is also shown by size exclusion chromatography in which Abeta(1)(-)(40)-Lactam(D23/K28) forms oligomers even at concentrations of approximately 1-5 microM. Under the same conditions, Abeta(1)(-)(40) shows no detectable oligomers by size exclusion chromatography. Our data suggest that Abeta(1)(-)(40)-Lactam(D23/K28) could bypass an unfavorable folding step in fibrillogenesis, because the lactam linkage "preforms" a bendlike structure in the peptide. Consistent with this view Abeta(1)(-)(40) growth is efficiently nucleated by Abeta(1)(-)(40)-Lactam(D23/K28) fibril seeds. PMID- 15835890 TI - Solution structure and lipid membrane partitioning of VSTx1, an inhibitor of the KvAP potassium channel. AB - VSTx1 is a voltage sensor toxin from the spider Grammostola spatulata that inhibits KvAP, an archeabacterial voltage-activated K(+) channel whose X-ray structure has been reported. Although the receptor for VSTx1 and the mechanism of inhibition are unknown, the sequence of the toxin is related to hanatoxin (HaTx) and SGTx, two toxins that inhibit eukaryotic voltage-activated K(+) channels by binding to voltage sensors. VSTx1 has been recently shown to interact equally well with lipid membranes that contain zwitterionic or acidic phospholipids, and it has been proposed that the toxin receptor is located within a region of the channel that is submerged in the membrane. As a first step toward understanding the inhibitory mechanism of VSTx1, we determined the three-dimensional solution structure of the toxin using NMR. Although the structure of VSTx1 is similar to HaTx and SGTx in terms of molecular fold and amphipathic character, the detailed positions of hydrophobic and surrounding charged residues in VSTx1 are very different than what is seen in the other toxins. The amphipathic character of VSTx1, notably the close apposition of basic and hydrophobic residues on one face of the toxin, raises the possibility that the toxin interacts with interfacial regions of the membrane. We reinvestigated the partitioning of VSTx1 into lipid membranes and find that VSTx1 partitioning requires negatively charged phospholipids. Intrinsic tryptophan fluorescence and acrylamide quenching experiments suggest that tryptophan residues on the hydrophobic surface of VSTx1 have a diminished exposure to water when the toxin interacts with membranes. The present results suggest that if membrane partitioning is involved in the mechanism by which VSTx1 inhibits voltage-activated K(+) channels, then binding of the toxin to the channel would likely occur at the interface between the polar headgroups and the hydrophobic phase of the membrane. PMID- 15835891 TI - Comparison of the global structure and dynamics of native and unmodified tRNAval. AB - The effects of post-transcriptional modifications on the structure and dynamics of Escherichia coli tRNA(val) were studied by NMR spectroscopy. NMR chemical shift data and residual dipolar couplings were used to show that the local secondary and tertiary structures are very similar in native and unmodified tRNA(val). Rigid body restrained molecular dynamics calculations showed that the global structure of tRNA is unchanged by the post-transcriptional modifications. Deuterium exchange NMR experiments were used to probe the dynamics and flexibility of native and unmodified tRNA(val). A similar set of slowly exchanging (t(1/2) > 3 min) imino protons were observed in both tRNAs, but the rates of exchange for the slowest exchanging imino protons were approximately 20 times faster in unmodified than in native tRNA. These results demonstrate that the dynamics and flexibility of tRNA(val), but not the local or global structure, are significantly affected by post-transcriptional modifications. PMID- 15835892 TI - Different effects of 4-hydroxyproline and 4-fluoroproline on the stability of collagen triple helix. AB - Differential scanning calorimetry (DSC) analyses of a series of collagen model peptides suggest that 4-hydroxyproline (Hyp) and 4-fluoroproline (fPro) have different effects on the stability of the collagen triple helices according to the sequence of amino acids and stereochemistry at the 4 positions of these imino acids. The thermodynamic parameters indicate that the enhanced stabilities are classified into two different types: the enthalpy term is primarily responsible for the enhanced stability of the triple helix of (Pro-Hyp(R)-Gly)(10), whereas the entropy term dominates the enhanced stability of (Pro-fPro(R)-Gly)(10). The difference between the molecular volumes observed in solution and intrinsic molecular volumes calculated from the crystal structure indicates the different hydration states of these peptides. (Pro-Hyp(R)-Gly)(10) is highly hydrated compared to (Pro-Pro-Gly)(10), which contributes to the larger enthalpy. In contrast, the volume of (Pro-fPro(R)-Gly)(10) shows a smaller degree of hydration than that of (Pro-Pro-Gly)(10). The entropic cost of forming the triple helix of the fPro-containing peptides is compensated by a decrease in an ordered structure of water molecules surrounding the peptide molecule, although the contribution of enthalpy originating from the hydration is reduced. These arguments about the different contribution of entropic and enthalpic terms were successfully applied to interpret the stability of the triple helix of (fPro(S)-Pro-Gly)(10) as well. PMID- 15835893 TI - Spiroiminodihydantoin lesions derived from guanine oxidation: structures, energetics, and functional implications. AB - Reactive oxygen species present in the cell generate DNA damage. One of the major oxidation products of guanine in DNA, 8-oxo-7,8-dihydroguanine, formed by loss of two electrons, is among the most extensively studied base lesions. The further removal of two electrons from this product can yield spiroiminodihydantoin (Sp) R and S stereoisomers. Both in vitro and in vivo experiments have shown that the Sp stereoisomers are highly mutagenic, causing G --> T and G --> C transversions. Hence, they are of interest as examples of endogenous DNA damage that may initiate cancer. To interpret the mutagenic properties of the Sp lesions, an understanding of their structural properties is needed. To elucidate these structural effects, we have carried out computational investigations at the level of the Sp-modified base and nucleoside. At the base level, quantum mechanical geometry optimization studies have revealed exact mirror image symmetry of the R and S stereoisomers, with a near-perpendicular geometry of the two rings. At the nucleoside level, an extensive survey of the potential energy surface by molecular mechanics calculations using AMBER has provided three-dimensional potential energy maps. These maps reveal that the range and flexibility of the glycosidic torsion angles are significantly more restricted in both stereoisomeric adducts than in unmodified 2'-deoxyguanosine. The structural and energetic results suggest that the unusual geometric, steric, and hydrogen bonding properties of these lesions underlie their mutagenicity. In addition, stereoisomer-specific differences indicate the possibility that their processing by cellular replication and repair enzymes may be differentially affected by their absolute configuration. PMID- 15835894 TI - Molecular visualization of the yeast Dmc1 protein ring and Dmc1-ssDNA nucleoprotein complex. AB - Saccharomyces cerevisiae Dmc1, a meiosis-specific homologue of RecA, catalyzes homologous pairing and strand exchange during meiotic DNA recombination. The purified budding yeast Dmc1 (ScDmc1) protein exhibits much weaker recombinase activity in vitro as compared to that of the Escherichia coli RecA protein. Using atomic force microscopy (AFM) with carbon nanotube tips, we found ScDmc1 forms rings with an external diameter of 18 nm and a central cavity of 4 nm. In the presence of single-stranded DNA (ssDNA), the majority of the ScDmc1 protein (90%) bound DNA as protein rings; only a small faction (10%) was able to form filamentous structure. In contrast, nearly all RecA proteins form fine helical nucleoprotein filaments with ssDNA under identical conditions. RecA-mediated recombinase activity is initiated through the nucleation of RecA onto ssDNA to form helical nucleoprotein filaments. Our results support the notion that ScDmc1 becomes catalytically active only when it forms a helical nucleoprotein filament with ssDNA. PMID- 15835895 TI - Solution structural study of a DNA duplex containing the guanine-N7 adduct formed by a cytotoxic platinum-acridine hybrid agent. AB - [PtCl(en)(ACRAMTU-S)](NO(3))(2) (PT-ACRAMTU; en = ethane-1,2-diamine, ACRAMTU = 1 [2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea) is a dual metalating/intercalating DNA binding drug conjugate that shows cytotoxicity at micromolar to nanomolar concentrations in a wide range of solid tumor cell lines. In approximately 80% of its adducts, PT-ACRAMTU binds to guanine-N7 in the major groove, selectively at 5'-CG sites [Budiman, M. E. et al. (2004) Biochemistry 43, 8560-8567]. Here, we report the synthesis, physical characterization, and NMR solution structure of a site-specifically modified octamer containing this adduct, 5'-CCTCGTCC-3'/3'-GGAGCAGG-5', where the asterisk indicates the [Pt(en)ACRAMTU)](3+) fragment. The structure was determined by a combination of high-resolution 2-D NMR spectroscopy and restrained molecular dynamics/molecular mechanics (rMD/MM) calculations using 179 NOE distance restraints and refined to an r(6) weighted residual (R(x)) of 9.2 x 10(-)(2) using the complete relaxation matrix approach. An average structure was calculated from the final ensemble of 19 rMD geometries showing pairwise root-mean-square deviations of <1.05 A. The dual binding increases the thermal stability of the octamer compared to the unmodified duplex (DeltaT(m) = 13.2 degrees ). The modified sequence shows structural features reminiscent of both B- and A-type DNA. Watson-Crick hydrogen bonding is intact at and beyond the adduct site. Platinum is bound to the N7 position of G5 in the major groove, and ACRAMTU intercalates into the central 5' C4G5/C12G13 base-pair step on the 5'-face of the platinated nucleobase. The chromophore's long axis is aligned with the long axes of the adjacent base pairs, maximizing intermolecular pi-pi stacking interactions. PT-ACRAMTU lengthens (rise, 6.62 A) and unwinds (twist, 15.4 degrees ) the duplex at the central base pair step but does not cause helical bending. No C3'-endo deoxyribose pucker and no significant roll are observed at the site of intercalation/platination, which clearly distinguishes the PT-ACRAMTU-induced damage from the 1,2-intrastrand cross-link formed by cisplatin. Overall, the DNA perturbations produced by PT ACRAMTU do not appear to mimic those caused by the major cisplatin lesion. Instead, intriguing structural similarities are observed for PT-ACRAMTU's monoadduct and the N7 adducts of dual major-groove alkylating/intercalating antitumor agents, such as the pluramycins. PMID- 15835896 TI - Two nuclear localization signals required for transport from the cytosol to the nucleus of externally added FGF-1 translocated into cells. AB - Externally added FGF-1 is transported into the nucleus of cells. It was earlier shown that FGF-1 contains an N-terminal nuclear localization signal (NLS) implicated in the stimulation of DNA synthesis. We here provide evidence that FGF 1 contains a second putative NLS (NLS2), which is located near the C-terminus. It is a bipartite NLS consisting of two clusters of lysines separated by a spacer of 10 amino acids. A fusion protein of GFP and the bipartite NLS was more efficiently transported into the nucleus than GFP alone, indicating that it can act as an NLS in the living cell. FGF-1 mutated in the N-terminal NLS (NLS1) or in the first cluster of the bipartite NLS2 bound to heparin and FGF receptors and activated downstream signaling similarly to the wild-type growth factor. Mutations in the second cluster of NLS2 resulted in impaired interaction with heparin and reduced stability. When radiolabeled FGF-1 with mutated NLS1 or the first lysine cluster of NLS2 was added to NIH/3T3 cells, it was translocated into the cytosol, but not transported efficiently to the nucleus. Phosphorylation of FGF-1 occurs normally in the nucleus, and while wild-type FGF-1 was phosphorylated after addition to cells, the NLS mutants were not. It therefore appears that both NLS1 and NLS2 are important for efficient transport of FGF-1 to the nucleus. Stimulation of DNA synthesis by FGF-1 with mutations in both NLSs was reduced considerably indicating that efficient transport to the nucleus may be involved in the stimulation of DNA synthesis. PMID- 15835897 TI - Role of aspartate 94 in the decay of the peroxide intermediate in the multicopper oxidase Fet3p. AB - Fet3p is a multicopper oxidase that contains four Cu ions: one type 1, one type 2, and a coupled binuclear type 3 site. The type 2 and type 3 centers form a trinuclear cluster that is the active site for O(2) reduction to H(2)O. When the type 1 Cu is depleted (C484S mutation), the reaction of the reduced trinuclear cluster with O(2) generates a peroxide intermediate. Kinetic studies of the decay of the peroxide intermediate suggest that a carboxyl residue (D94 in Fet3p) assists the reductive cleavage of the O-O bond at low pH. Mutations at the D94 residue (D94A, D94N, and D94E) have been studied to evaluate its role in the decay of the peroxide intermediate. Spectroscopic studies show that the D94 mutations affect the geometric and electronic structure of the trinuclear cluster in a way that is consistent with the hydrogen bond connectivity of D94. While the D94E mutation does not affect the initial reaction of the cluster with O(2), the D94A mutation causes larger structural changes that render the trinuclear cluster unreactive toward O(2), demonstrating a structural role for the D94 residue. The decay of the peroxide intermediate is markedly affected by the D94E mutation, confirming the involvement of D94 in this reaction. The D94 residue appears to activate a proton of the type 2 Cu(+)-bound water for participation in the transition state. These studies provide new insight into the role of D94 and proton involvement in the reductive cleavage of the O-O bond. PMID- 15835898 TI - Expression, mutagenesis, and characterization of recombinant low-potential cytochrome c550 of photosystem II. AB - Cytochrome c(550) of the photosystem II complex of cyanobacteria is an unusual member of the large protein family of monoheme c-type cytochromes. Despite the fact that it shares considerable amino acid sequence similarity and has a protein fold similar to the other members of the family, Cyt.c(550) has a midpoint potential (E(m7) = -250 mV) that is much lower than the positive midpoint potentials characteristic (E(m7) = 100-300mV) of this cytochrome family. An E. coli heterologous expression system involving secretion of the recombinant protein from Synechocystis PCC6803 to the periplasm was utilized to allow production of wild-type and mutant forms of the cytochrome. For most of the variants studied, the yield of protein was significantly enhanced by growth at 28 degrees C and inclusion of sucrose and betaine, in addition to isopropyl-beta-d thiogalactoside (IPTG), to the growth medium of the E. coli expression host. Analysis of the protein products revealed that the wild-type protein maintained the redox and visible spectroscopic characteristics of the authentic protein. Mutations in the residues engaging in hydrogen bond interactions with the heme propionate (Asn49) and the axial 6th ligand His92 (Pro93) resulted in small (12 20 mV), but reproducible, upshifts in midpoint redox potential. Substitution of the axial ligand His92 with Met produced no discernible changes in the optical spectrum relative to the wild-type despite the fact that in this mutant, unlike the others studied here, the thioether linkage either was not formed or was highly labile as evidenced by loss of the heme during SDS-PAGE. On the other hand, the midpoint potential of the C550-H92M mutant was upshifted by approximately 70 mV. This value is significantly less of a perturbation than that observed in a similar mutant that is natively expressed in Thermosynechoccocus but appears to have an intact thioether linkage between the heme and the polypeptide moiety. PMID- 15835899 TI - Crystallographic evidence for a new ensemble of ligand-induced allosteric transitions in hemoglobin: the T-to-T(high) quaternary transitions. AB - A detailed description of hemoglobin cooperativity requires knowledge of the dimer-dimer interactions responsible for the low ligand affinity of the quaternary-T tetramer, the "quaternary-T constraints", along with stereochemical pathways that specify how ligand binding disrupts these quaternary constraints. The recent mutagenic screen of Noble et al. [Noble, R. W., et al. (2001) Biochemistry 40, 12357-12368] has identified the major region of quaternary constraint to be a cluster of residues at the alpha1beta2 interface that is centered at Trp37beta. In this paper, crystallographic studies are presented for most of the mutant hemoglobins studied by Noble et al. These crystallographic experiments identify structural transitions-referred to as T-to-T(High) transitions-between the quaternary-T structure of wild-type deoxyhemoglobin and an ensemble of related T-like quaternary structures that are induced by some mutations in the Trp37beta cluster and/or by exposing crystals of wild-type or mutant deoxyhemoglobins to oxygen. The T-to-T(High) quaternary transitions consist of a rotation of the alpha1beta1 dimer relative to the alpha2beta2 dimer as well as a coupled alphabeta dimer bending component that consists of a small rotation of the alpha1 subunit relative to the beta1 subunit (and a symmetry related rotation of the alpha2 subunit relative to the beta2 subunit). In addition, differences in subunit tertiary structure associated with the T-to T(High) transitions suggest two stereochemical pathways (one associated with the alpha subunits and one associated with the betasubunits) by which ligand binding specifically disrupts quaternary constraints in the Trp37beta cluster. In the alpha subunits, ligand binding induces a shift of the heme iron producing tension in a chain of covalent bonds that extends from the Fe-N(epsilon)(2)His(F8)alpha1 bond to the peptide backbone bonds of residues His87(F8)alpha1 and Ala88(F9)alpha1. This tension induces an alpha-to-pi transition in the COOH terminal end of the F-helix that shifts the beta-carbon of Ala88alpha1 by approximately 1.5 A directly into the side chain of Tyr140alpha1 (a key residue in the Trp37beta2 cluster). Collectively these structural changes constitute a relatively short pathway by which ligand binding forces Tyr140alpha1 into the alpha1beta2 interface disrupting quaternary constraints associated with the Trp37beta2 cluster. In the beta subunits, our analysis suggests a more extended energy transduction pathway in which ligand-induced beta1-heme movement triggers tertiary changes in the beta1 subunit that promote alpha1beta1 dimer bending that disrupts quaternary constraints in the Trp37beta2 cluster at the alpha1beta2 interface. PMID- 15835900 TI - Effects of chromogranin expression on inositol 1,4,5-trisphosphate-induced intracellular Ca2+ mobilization. AB - We show here that expression of chromogranins in non-neuroendocrine NIH3T3 cells significantly increased the amount of IP(3)-mediated intracellular Ca(2+) mobilization in these cells, whereas suppression of them in neuroendocrine PC12 cells decreased the amount of mobilized Ca(2+). We have therefore investigated the relationship between the IP(3)-induced intracellular Ca(2+) mobilization and secretory granules. The level of IP(3)-mediated Ca(2+) release in CGA-expressing NIH3T3 cells was 40% higher than in the control cells, while that of CGB expressing cells was 134% higher, reflecting the number of secretory granules formed. Suppression of CGA and CGB expression in PC12 cells resulted in 41 and 78% reductions in the number of secretory granules, respectively, while the extents of IP(3)-induced Ca(2+) release in these cells were reduced 40 and 69%, respectively. The newly formed secretory granules of NIH3T3 cells contained all three isoforms of the IP(3)Rs. Comparison of the concentrations of the IP(3)R isoforms expressed in the ER and nucleus of chromogranin-expressing and nonexpressing NIH3T3 cells did not show significant differences, indicating that chromogranin expression did not affect the expression of endogenous IP(3)Rs. Nonetheless, the IP(3)R concentrations in secretory granules of chromogranin expressing NIH3T3 cells were 3.5-4.7-fold higher than those of the ER, similar to the levels found in secretory granules of neuroendocrine chromaffin cells, thus suggesting that the IP(3)Rs targeted to the newly formed secretory granules are newly induced by chromogranins without affecting the expression of intrinsic IP(3)Rs. These results strongly suggest that the extent of IP(3)-induced intracellular Ca(2+) mobilization in secretory cells is closely related to the number of secretory granules. PMID- 15835901 TI - Mass spectrometric analysis of agonist effects on posttranslational modifications of the beta-2 adrenoceptor in mammalian cells. AB - Posttranslational modifications (PTMs) of the beta-2 adrenoceptor (B2AR) play a fundamental role in receptor regulation by agonists. We have examined the effects of several agonists on net levels of B2AR palmitoylation and phosphorylation using epitope tagging in stably transfected human embryonal kidney (HEK) 293 cells, immunoaffinity purification, and mass spectrometry combined with the method of stable isotope labeling by amino acids in cell culture (SILAC). Palmitoylation of Cys341 was confirmed and did not change detectably after 30 min exposure of cells to saturating concentrations of dopamine, epinephrine, or isoproterenol. However, all of these agonists produced a marked increase in net phosphorylation. Phosphorylation of the third cytoplasmic loop was increased to a similar degree by all three agonists, whereas differences between agonists were observed in net phosphorylation of the carboxyl-terminal cytoplasmic domain (isoproterenol approximately epinephrine >> dopamine). Interestingly, agonist induced phosphorylation of the carboxyl-terminal cytoplasmic domain was observed exclusively in a proximal portion (between residues 339-369). None of the agonists produced detectable phosphorylation in a distal portion of the cytoplasmic tail, which contains all sites of agonist-induced phosphorylation identified previously by in vitro reconstitution. These results provide insight to agonist-dependent regulation of the B2AR in intact cells, suggest the existence of significant differences in regulatory phosphorylation events occurring between in vitro and in vivo conditions, and outline a general analytical approach to investigate regulated PTM of receptors in mammalian cells. PMID- 15835902 TI - Linker region of a halobacterial transducer protein interacts directly with its sensor retinal protein. AB - pHtrII, a pharaonis halobacterial transducer protein, possesses two transmembrane helices and forms a signaling complex with pharaonis phoborhodopsin (ppR, also called pharaonis sensory rhodopsin II, NpSRII) within the halobacterial membrane. This complex transmits a light signal to the sensory system located in the cytoplasm. It has been suggested that the linker region connecting the transmembrane region and the methylation region of pHtrII is important for binding to ppR and subsequent photosignal transduction. In this study, we present evidence to suggest that the linker region itself interacts directly with ppR in addition to the interaction in the membrane region. An in vitro pull-down assay revealed that the linker region bound to ppR, and its dissociation constant (K(D)) was estimated to be approximately 10 microM using isothermal titration calorimetry (ITC). Solution NMR analyses showed that ppR interacted with the linker region of pHtrII (pHtrII(G83)(-)(Q149)) and resulted in the broadening of many peaks, indicating structural changes within this region. These results suggest that the pHtrII linker region interacts directly with ppR. There was no demonstrable interaction between the C-terminal region of ppR (ppR(Gly224)( )(His247)) and either the linker region (pHtrII(G83)(-)(Q149)) or the transmembrane region (pHtrII(M1)(-)(E114)) of pHtrII. On the basis of the NMR, CD, and photochemical data, we discuss the structural changes and role of the linker region of pHtrII in relation to photosignal transduction. PMID- 15835903 TI - Molecular recognition of the substrate diphosphate group governs product diversity in trichodiene synthase mutants. AB - The X-ray crystal structures of Y305F trichodiene synthase and its complex with coproduct inorganic pyrophosphate (PP(i)) and of Y305F and D100E trichodiene synthases in ternary complexes with PP(i) and aza analogues of the bisabolyl carbocation intermediate are reported. The Y305F substitution in the basic D(302)RRYR motif does not cause large changes in the overall structure in comparison with the wild-type enzyme in either the uncomplexed enzyme or its complex with PP(i). However, the loss of the Y305F-PP(i) hydrogen bond appears to be compensated by a very slight shift in the position of the side chain of R304. The putative bisabolyl carbocation mimic, R-azabisabolene, binds in a conformation and orientation that does not appear to mimic that of the actual carbocation intermediate, suggesting that the avid inhibition by R- and S azabisabolenes arises more from favorable electrostatic interactions with PP(i) rather than any special resemblance to a reaction intermediate. Greater enclosed active-site volumes result from the Y305F and D100E mutations that appear to confer greater variability in ligand-binding conformations and orientations, which results in the formation of aberrant cyclization products. Because the binding conformations and orientations of R-azabisabolene to Y305F and D100E trichodiene synthases do not correspond to binding conformations required for product formation and because the binding conformations and orientations of diverse substrate and carbocation analogues to other cyclases such as 5-epi aristolochene synthase and bornyl diphosphate synthase generally do not correspond to catalytically productive complexes, we conclude that the formation of transient carbocation intermediates in terpene cyclization reactions is generally under kinetic rather than thermodynamic control. PMID- 15835904 TI - Intermediates and transition states in thiamin diphosphate-dependent decarboxylases. A kinetic and NMR study on wild-type indolepyruvate decarboxylase and variants using indolepyruvate, benzoylformate, and pyruvate as substrates. AB - The thiamin diphosphate (ThDP)-dependent enzyme indolepyruvate decarboxylase (IPDC) is involved in the biosynthetic pathway of the phytohormone 3-indoleacetic acid and catalyzes the nonoxidative decarboxylation of 3-indolepyruvate to 3 indoleacetaldehyde and carbon dioxide. The steady-state distribution of covalent ThDP intermediates of IPDC reacting with 3-indolepyruvate and the alternative substrates benzoylformate and pyruvate has been analyzed by (1)H NMR spectroscopy. For the first time, we are able to isolate and directly assign covalent intermediates of ThDP with aromatic substrates. The intermediate analysis of IPDC variants is used to infer the involvement of active site side chains and functional groups of the cofactor in distinct catalytic steps during turnover of the different substrates. As a result, three residues (glutamate 468, aspartate 29, and histidine 115) positioned perpendicular to the thiazolium moiety of ThDP are involved in binding of all substrates and decarboxylation of the respective tetrahedral ThDP-substrate adducts. Most likely, interactions of these side chains with the substrate-derived carboxylate account for an optimal orientation of the substrate and/or intermediate in the course of carbon-carbon ligation and decarboxylation supporting the suggested least-motion, maximum overlap mechanism. The active site residue glutamine 383, which is located at the opposite site of the thiazolium nucleus as the "carboxylate pocket" (formed by the Glu-Asp-His triad), is central to the substrate specificity of IPDC, probably through orbital alignment. The Glu51-cofactor proton shuttle is, conjointly with the Glu-Asp-His triad, involved in multiple proton transfer steps, including ylide generation, substrate binding, and product release. Studies with para substituted benzoylformate substrates demonstrate that the electronic properties of the substrate affect the stabilization or destabilization of the carbanion intermediate or carbanion-like transition state and in that way alter the rate dependence on decarboxylation. In conclusion, general mechanistic principles of catalysis of ThDP-dependent enzymes are discussed. PMID- 15835906 TI - The N-terminal domain of MYO18A has an ATP-insensitive actin-binding site. AB - Myosin XVIII is the recently identified 18th class of myosins, and its members are composed of a unique N-terminal domain, a motor domain with an unusual sequence around the ATPase site, one IQ motif, a segmented coiled-coil region for dimerization, and a C-terminal globular tail. To gain insight into the functions of this unique myosin, we characterized its human homologue, MYO18A, focusing on the functional roles of the characteristic N-terminal domain that contains a PDZ module known to mediate protein-protein interaction. GFP-tagged full-length and C terminally truncated MYO18A molecules that were expressed in HeLa cells exhibited colocalization with actin filaments. Chemical cross-linking of these molecules showed that they form stable dimers as expected from their putative coiled-coil tails. Cosedimentation of the various types of truncated MYO18A constructs with actin filaments indicated the presence of an ATP-insensitive actin-binding site in the N-terminal domain. Further studies on truncated constructs of the N terminal domain indicated that this actin-binding site is located outside the PDZ module, but within the middle region of this domain, which does not show any homology with the known actin-binding motifs. These results imply that this dimeric myosin might stably cross-link actin filaments by two ATP-insensitive actin-binding sites at the N-terminal domains for higher-order organization of the actin cytoskeleton. PMID- 15835905 TI - The structure of the Candida albicans Ess1 prolyl isomerase reveals a well ordered linker that restricts domain mobility. AB - Ess1 is a peptidyl-prolyl cis/trans isomerase (PPIase) that binds to the carboxy terminal domain (CTD) of RNA polymerase II. Ess1 is thought to function by inducing conformational changes in the CTD that control the assembly of cofactor complexes on the transcription unit. Ess1 (also called Pin1) is highly conserved throughout the eukaryotic kingdom and is required for growth in some species, including the human fungal pathogen Candida albicans. Here we report the crystal structure of the C. albicansEss1 protein, determined at 1.6 A resolution. The structure reveals two domains, the WW and the isomerase domain, that have conformations essentially identical to those of human Pin1. However, the linker region that joins the two domains is quite different. In human Pin1, this linker is short and flexible, and part of it is unstructured. In contrast, the fungal Ess1 linker is highly ordered and contains a long alpha-helix. This structure results in a rigid juxtaposition of the WW and isomerase domains, in an orientation that is distinct from that observed in Pin1, and that eliminates a hydrophobic pocket between the domains that was implicated as the main substrate recognition site. These differences suggest distinct modes of interaction with long substrate molecules, such as the CTD of RNA polymerase II. We also show that C. albicans ess1(-)() mutants are attenuated for in vivo survival in mice. Together, these results suggest that CaEss1 might constitute a useful antifungal drug target, and that structural differences between the fungal and human enzymes could be exploited for drug design. PMID- 15835907 TI - Chemistry of the catalytic conversion of phthalate into its cis-dihydrodiol during the reaction of oxygen with the reduced form of phthalate dioxygenase. AB - The phthalate dioxygenase system, a Rieske non-heme iron dioxygenase, catalyzes the dihydroxylation of phthalate to form the 4,5-dihydro-cis-dihydrodiol of phthalate (DHD). It has two components: phthalate dioxygenase (PDO), a multimer with one Rieske-type [2Fe-2S] and one mononuclear Fe(II) center per monomer, and a reductase (PDR) that contains flavin mononucleotide (FMN) and a plant-type ferredoxin [2Fe-2S] center. This work shows that product formation in steady state reactions is tightly coupled to electron delivery, with 1 dihydrodiol (DHD) of phthalate formed for every 2 electrons delivered from NADH. However, in reactions of reduced PDO with O(2), only about 0.5 DHD is formed per Rieske center that becomes oxidized. Although the product forms rapidly, its release from PDO is slow in these reactions with oxygen that do not include reductase and NADH. EPR data show that, at the completion of the oxidation, iron in the mononuclear center remains in the ferrous state. In contrast, naphthalene dioxygenase (NDO) [Wolfe, M. D., Parales, J. V., Gibson, D. T., and Lipscomb, J. D. (2001) J. Biol. Chem. 276, 1945-1953] and benzoate dioxygenase (BZDO) [Wolfe, M. D., Altier, D. J., Stubna, A., Popescu, C. V., Munck, E., and Lipscomb, J. D. (2002) Biochemistry, 41, 9611-9626], related Rieske non-heme iron dioxygenases, form 1 DHD per Rieske center oxidized, and the mononuclear center iron ends up ferric. Thus, both electrons from reduced NDO and BZDO monomers are used to form the product, whereas only the reduced Rieske centers in PDO become oxidized during production of DHD. This emphasizes the importance of PDO subunit interaction in catalysis. Electron redistribution was practically unaffected by the presence of oxidized PDR. A scheme is presented that emphasizes some of the differences in the mechanisms involved in substrate hydroxylation employed by PDO and either NDO or BZDO. PMID- 15835908 TI - Saturation transfer difference measurements with SU(VAR)3-9 and S-adenosyl-L methionine. AB - Saturation transfer difference NMR measurements were performed to investigate the interaction of S-adenosyl-l-methionine (AdoMet) with SU(VAR)3-9 from Drosophila melanogaster. SU(VAR)3-9 has a SET domain and plays an important role in methylation of lysine-9 of histone H3 which results in gene silencing. We determined the binding epitope of AdoMet and compared it with a crystal structure of another SET protein. PMID- 15835909 TI - Molecular crosstalk between the nucleotide specificity determinant of the SRP GTPase and the SRP receptor. AB - In signal recognition particle (SRP)-dependent targeting of proteins to the bacterial plasma membrane, two GTPases, Ffh (the SRP GTPase) and FtsY (the receptor GTPase), form a complex in which both proteins reciprocally stimulate each other's GTPase activities. We mutated Asp251 in the Ffh active site to Asn (D251N), converting Ffh to a xanthosine 5'-triphosphate (XTP)-specific protein as has been observed in many other GTPases. Unexpectedly, mutant SRP(D251N) is severely compromised in the formation of an active SRP.FtsY complex when bound with cognate XTP, and even more surprisingly, mutant SRP(D251N) works better when bound with noncognate GTP. These paradoxical results are explained by a model in which Ffh Asp251 forms a bidentate interaction with not only the bound GTP but also the receptor FtsY across the dimer interface. These interactions form part of the network that seals the lateral entrance to the composite active site at the dimer interface, thereby ensuring the electrostatic and/or structural integrity of the active site and contributing to the formation of an active SRP.FtsY complex. PMID- 15835910 TI - Mechanistic studies of the oxygen-mediated oxidation of nitrosylhemoglobin. AB - Nitrosylhemoglobin (HbFe(II)NO) has been shown to be generated in vivo from the reaction of deoxyHb with NO(*) as well as with nitrite. Despite the physiological importance attributed to this form of Hb, its reactivity has not been investigated in detail. In this study, we showed that the rate of oxidation of HbFe(II)NO by O(2) does not depend on the O(2) concentration. The reaction time courses had to be fitted to a two-exponential expression, and the obtained rates were approximately 2 x 10(-)(4) and 1 x 10(-)(4) s(-)(1), respectively. In the presence of the allosteric effector inositol hexaphosphate (IHP), the value for the fast component of the rate was significantly larger (44 x 10(-)(4) s(-)(1)) whereas that for the slow step was only slightly higher (2.5 x 10(-)(4) s(-)(1)). Moreover, we found that both in the absence and in the presence of IHP the rate of the O(2)-mediated oxidation of HbFe(II)NO is essentially identical to that of NO(*) dissociation from HbFe(II)NO, determined under analogous conditions by replacement of NO(*) with CO in the presence of an excess of dithionite. Taken together, our data show that the reaction between O(2) and HbFe(II)NO proceeds in three steps via dissociation of NO(*) (rate-determining step), binding of O(2) to deoxyHb, and NO(*)-mediated oxidation of oxyHb to metHb and nitrate. PMID- 15835911 TI - Implications of the effects of viscosity, macromolecular crowding, and temperature for the transient interaction between cytochrome f and plastocyanin from the cyanobacterium Phormidium laminosum. AB - The reaction between cytochrome f and plastocyanin is a central feature of the photosynthetic electron-transport system of all oxygenic organisms. We have studied the reaction in solution to understand how the very weak binding between the two proteins from Phormidium laminosum can nevertheless lead to fast rates of electron transfer. In a previous publication [Schlarb-Ridley, B. G., et al. (2003) Biochemistry 42, 4057-4063], we suggested that the reaction is diffusion controlled because of a strong effect of viscosity of the medium. The effects of viscosity and temperature have now been examined in detail. High molecular mass viscogens (Ficoll 70 and Dextran 70), which might mimic in vivo conditions, had little effect up to a relative viscosity of 4. Low molecular mass viscogens (ethane diol, glycerol, and sucrose) strongly decreased the bimolecular rate constant (k(2)) over a similar viscosity range. The effects correlated well with the viscosities of the solutions of the three reagents but not with their dielectric constants or molalities. A power law dependence of k(2) on viscosity suggested that k(2) depends on two viscosity-sensitive reactions in series, while the reverse reactions are little affected by viscosity. The results were incompatible with diffusion control of the overall reaction. Determination of the effect of temperature on k(2) gave an activation enthalpy, DeltaH(++) = 45 kJ mol(-)(1), which is also incompatible with diffusion control. The results were interpreted in terms of a model in which the stable form of the protein-protein complex requires further thermal activation to be competent for electron transfer. PMID- 15835912 TI - The catalytically active serine protease domain of human complement factor I. AB - Factor I (fI) is a major regulator of complement. As a protease it has very restricted specificity, cleaving only C3b or C4b in the presence of a cofactor such as factor H (fH). Cleavage of C3b by fI yields iC3b, a major opsonin. The cleavage occurs through the formation of a ternary complex between the enzyme, the substrate, and the cofactor. The catalytic subunit of fI, the SP domain, accommodates substrate recognition and cleavage. The role of the fI heavy chain within the catalysis complex is unknown. Using partial proteolysis and affinity chromatography an intact form of the SP domain was generated and isolated from fI in high yield. fI and the SP domain were found to have similar amidolytic activities but strikingly different proteolytic activities on C3(NH(3)). fI did not cleave C3(NH(3)) in the absence of fH, while in its presence it cleaved C3(NH(3)) rapidly at two sites. The SP domain, however, slowly cleaved C3(NH(3)) in the absence of fH, at more than two sites. Cleavage by the SP domain was inhibited, not stimulated, by fH. Pefabloc SC and antipain inhibited the proteolytic activity of both fI and the SP domain, but suramin inhibited only fI and not the SP domain. The contrast in the proteolytic activities suggests that the heavy chain domains and the cofactor must have roles in orienting the natural substrates and restricting cleavage to the two sites which yield iC3b through a highly specific catalysis. PMID- 15835913 TI - Characterization of the disulfide bonds and free cysteine residues of the Chlamydia trachomatis mouse pneumonitis major outer membrane protein. AB - Members of the genus Chlamydia lack a peptidoglycan layer. As a substitute for peptidoglycan, it has been proposed that several cysteine rich proteins, including the major outer membrane protein (MOMP), form disulfide bonds to provide rigidity to the cell wall. Alignment of the amino acids sequences of the MOMP from various serovars of Chlamydia showed that they have from 7 to 10 cysteine residues and seven of them are highly conserved. Which of these are free cysteine residues and which are involved in disulfide bonds is unknown. The complexity of the outer membrane of Chlamydia precludes at this point the characterization of the structure of the cysteines directly in the bacteria. Therefore, mass spectrometric analysis of a purified and refolded MOMP was used in this study. Characterization of the structure of this preparation of the MOMP is critical because it has been shown, in an animal model, to be a very effective vaccine against respiratory and genital infections. Here, we demonstrated that in this MOMP preparation four cysteines are involved in disulfide bonds, with intramolecular pairs formed between Cys(48) and Cys(55) and between Cys(201) and Cys(203). A stepwise alkylation, reduction, alkylation process using two different alkylating reagents was required to establish the Cys(48)-Cys(55) disulfide pair. The other residues in MOMP, Cys(51), Cys(136), Cys(226), and Cys(351), are free cysteines and could potentially form disulfide-linked complexes with other MOMP or other membrane proteins. PMID- 15835914 TI - Characterization of structure and metal ions specificity of Co2+-binding RNA aptamers. AB - Studies on RNA motifs capable of binding metal ions have largely focused on Mg(2+)-specific motifs, therefore information concerning interactions of other metal ions with RNA is still very limited. Application of the in vitro selection approach allowed us to isolate two RNA aptamers that bind Co(2+) ions. Structural analysis of their secondary structures revealed the presence of two motifs, loop E and "kissing" loop complex, commonly occurring in RNA molecules. The Co(2+) induced cleavage method was used for identification of Co(2+)-binding sites after the determination of the optimal cleavage conditions. In the aptamers, Co(2+) ions seem to bind to N7 atoms of purines, inducing cleavage of the adjacent phosphodiester bonds, similarly as is the case with yeast tRNA(Phe). Although the in vitro selection experiment was carried out in the presence of Co(2+) ions only, the aptamers displayed broader metal ions specificity. This was shown by inhibition of Co(2+)-induced cleavages in the presence of the following transition metal ions: Zn(2+), Cd(2+), Ni(2+), and Co(NH(3))(6)(3+) complex. On the other hand, alkaline metal ions such as Mg(2+), Ca(2+), Sr(2+), and Ba(2+) affected Co(2+)-induced cleavages only slightly. Multiple metal ions specificity of Co(2+)-binding sites has also been reported for other in vitro selected or natural RNAs. Among many factors that influence metal specificity of the Co(2+) binding pocket, chemical properties of metal ions, such as their hardness as well as the structure of the coordination site, seem to be particularly important. PMID- 15835915 TI - Urea promotes polyproline II helix formation: implications for protein denatured states. AB - It is commonly assumed that urea denatures proteins by promoting backbone disorder, resulting in random-coil behavior. Indeed, it has been demonstrated that highly denatured proteins obey random-coil statistics. However, the random coil model is specified by the global geometric properties of a polymeric chain and does not preclude locally ordered backbone structure. While urea clearly disfavors a compact native structure, it is not clear that the resulting backbone conformations are disordered. Using circular dichroism (CD) spectroscopy, we demonstrate that urea promotes formation of left-handed polyproline II (P(II)) helical structures in both short peptides and denatured proteins. The observed increase in P(II) content is sequence-dependent. These data indicate that denatured states possess significant amounts of locally ordered backbone structure. It is time for the formulation of new denatured-state models that take into account the presence of significant local backbone structure. Criteria for such models are outlined. PMID- 15835916 TI - Kruppel-like factor 2 transcriptional regulation involves heterogeneous nuclear ribonucleoproteins and acetyltransferases. AB - Kruppel-like factor 2 (KLF2) is expressed in several cell types, and knockout animals have shown that KLF2 gene regulation is involved in multiple biological processes. These include maintaining T-cells in the quiescent state, preventing preadipocytes from differentiating into mature adipocytes, stabilizing blood vessel walls through endothelial cell function, and advancing the later stages of lung development. Defining the regulation of KLF2 expression is important to understand these diverse functions. Promoter analysis of KLF2 has revealed that a region between -138 and -111 base pairs is required for its transcription, and this nucleotide sequence occurs in a region that is highly conserved in evolution. The present study was carried out to identify transcription factors that bind to this region of the KLF2 promoter. Nuclear factors were enriched by DNA affinity chromatography using the conserved nucleotide sequence of the KLF2 promoter. Mass spectrometry analysis of the proteins eluted from the affinity matrix identified several proteins, including glucose regulated protein-78 kDa (GRP-78), heterogeneous nuclear ribonucleoprotein (hnRNP)-U, hnRNP-D, CArG binding factor (CBF), P300/CBP associated factor (PCAF), cAMP response element binding protein (CREB) and SWI/SNF. The binding of these proteins to the highly conserved region of the KLF2 promoter element was tested by electrophoretic mobility supershift assays and chromatin immunoprecipitation analysis. These procedures confirmed that hnRNP-U, hnRNP-D, PCAF, and P-300 bind to the KLF2 promoter. Transactivation experiments demonstrated that these proteins are important for regulating KLF2 transcription. Of special interest is the role of hnRNPs in the transcription of the KLF2 gene. PMID- 15835917 TI - Phosphorylation of the human Fhit tumor suppressor on tyrosine 114 in Escherichia coli and unexpected steady state kinetics of the phosphorylated forms. AB - The human tumor suppressor Fhit is a homodimeric histidine triad (HIT) protein of 147 amino acids which has Ap(3)A hydrolase activity. We have recently discovered that Fhit is phosphorylated in vivo and is phosphorylated in vitro by Src kinase [Pekarsky, Y., Garrison, P. N., Palamarchuk, A., Zanesi, N., Aqeilan, R. I., Huebner, K., Barnes, L. D., and Croce, C. M. (2004) Proc. Natl. Acad. Sci. U.S.A. 101, 3775-3779]. Now we have coexpressed Fhit with the elk tyrosine kinase in Escherichia coli to generate phosphorylated forms of Fhit. Unphosphorylated Fhit, Fhit phosphorylated on one subunit, and Fhit phosphorylated on both subunits were purified to apparent homogeneity by column chromatography on anion-exchange and gel filtration resins. MALDI-TOF and HPLC-ESI tandem mass spectrometry of intact Fhit and proteolytic peptides of Fhit demonstrated that Fhit is phosphorylated on Y(114) on either one or both subunits. Monophosphorylated Fhit exhibited monophasic kinetics with K(m) and k(cat) values approximately 2- and approximately 7-fold lower, respectively, than the corresponding values for unphosphorylated Fhit. Diphosphorylated Fhit exhibited biphasic kinetics. One site had K(m) and k(cat) values approximately 2- and approximately 140-fold lower, respectively, than the corresponding values for unphosphorylated Fhit. The second site had a K(m) approximately 60-fold higher and a k(cat) approximately 6 fold lower than the corresponding values for unphosphorylated Fhit. The unexpected kinetic patterns for the phosphorylated forms suggest the system may be enzymologically novel. The decreases in the values of K(m) and k(cat) for the phosphorylated forms in comparison to those of unphosphorylated Fhit favor the formation and lifetime of the Fhit-Ap(3)A complex, which may enhance the tumor suppressor activity of Fhit. PMID- 15835918 TI - Tandem mass spectrometry for the examination of the posttranslational modifications of high-mobility group A1 proteins: symmetric and asymmetric dimethylation of Arg25 in HMGA1a protein. AB - High-mobility group (HMG) A1a and A1b proteins are among a family of HMGA proteins that bind to the minor groove of AT-rich regions of DNA. Here we employed tandem mass spectrometry and determined without ambiguity the sites of phosphorylation and the nature of methylation of HMGA1 proteins that were isolated from the PC-3 human prostate cancer cells. We showed by LC-MS/MS that Ser101 and Ser102 were completely phosphorylated in HMGA1a protein, whereas only a portion of the protein was phosphorylated at Ser98. We also found that the HMGA1b protein was phosphorylated at the corresponding sites, that is, Ser90, Ser91 and Ser87. In addition, Arg25, which is within the first DNA-binding AT hook domain of HMGA1a, was both mono- and dimethylated. Moreover, both symmetric and asymmetric dimethylations were observed. The closely related HMGA1b protein, however, was not methylated. The unambiguous identification of the sites of phosphorylation and the nature of methylation facilitates the future examination of the biological implications of the HMGA1 proteins. PMID- 15835919 TI - Dimerization of the exocyst protein Sec6p and its interaction with the t-SNARE Sec9p. AB - Vesicles in eukaryotic cells transport cargo between functionally distinct membrane-bound organelles and the plasma membrane for growth and secretion. Trafficking and fusion of vesicles to specific target sites are highly regulated processes that are not well understood at the molecular level. At the plasma membrane, tethering and fusion of secretory vesicles require the exocyst complex. As a step toward elucidation of the molecular architecture and biochemical function(s) of the exocyst complex, we expressed and purified the exocyst subunit Sec6p and demonstrated that it is a predominantly helical protein. Biophysical characterization of purified Sec6p by gel filtration and analytical ultracentrifugation experiments revealed that Sec6p is a dimer. Limited proteolysis defined an independently folded C-terminal domain (residues 300-805) that equilibrated between a dimer and monomer in solution. Removal of residues 300-410 from this construct yielded a well-folded, monomeric domain. These results demonstrate that residues 300-410 are necessary for dimerization, and the presence of the N-terminal region (1-299) increases dimer stability. Moreover, we found that the dimer of Sec6p binds to the plasma membrane t-SNARE Sec9p and inhibits the interaction between Sec9p and its partner t-SNARE Sso1p. This direct interaction between the exocyst complex and the t-SNARE implicates the exocyst in SNARE complex regulation. PMID- 15835920 TI - SHP represses transcriptional activity via recruitment of histone deacetylases. AB - The orphan receptor short heterodimer partner (SHP) is a common partner for a great number of nuclear receptors, and it plays an important role in many diverse physiological events. In a previous study, we described SHP as a strong repressor of the androgen receptor (AR). Herein, we addressed the mechanism of action of its negative activity on transcription. We first investigated the intrinsic repressive potential of SHP and mapped two core repressive domains to the amino acids 170-210 and 210-240. From GST pull-down assays, we demonstrated a direct interaction between SHP and diverse histone deacetylases (HDACs) as well as a strong interaction between HDAC1 and SHP inhibitory domains. We further supported the evidence for an interaction between SHP and HDAC1 by showing their co immunoprecipitation and provided evidence for the existence of a ternary complex comprising AR, SHP, and HDAC1. The use of trichostatin A (TSA), a specific inhibitor of HDAC activity, confirmed that HDACs significantly contribute to the intrinsic transrepressive activity of SHP. Finally, we showed that TSA reversed SHP-induced repression of AR, further emphasizing the relevance of the interaction between SHP and HDACs. This latter action affected in a very similar manner SHP-mediated repression of estrogen receptor alpha (ERalpha) transactivation. Altogether, our results indicate that SHP mediates most of its repressive effect through recruitment of HDACs and suggest that the physiological actions of SHP could be affected by HDAC inhibitors. PMID- 15835921 TI - Large enhancement of functional activity of active site-inhibited factor VIIa due to protein dimerization: insights into mechanism of assembly/disassembly from tissue factor. AB - Active site-inhibited blood clotting factor VIIa (fVIIai) binds to tissue factor (TF), a cell surface receptor that is exposed upon injury and initiates the blood clotting cascade. FVIIai blocks binding of the corresponding enzyme (fVIIa) or zymogen (fVII) forms of factor VII and inhibits coagulation. Although several studies have suggested that fVIIai may have superior anticoagulation effects in vivo, a challenge for use of fVIIai is cost of production. This study reports the properties of dimeric forms of fVIIai that are cross-linked through their active sites. Dimeric wild-type fVIIai was at least 75-fold more effective than monomeric fVIIai in blocking fVIIa association with TF. The dimer of a mutant fVIIai with higher membrane affinity was 1600-fold more effective. Anticoagulation by any form of fVIIai differed substantially from agents such as heparin and showed a delayed mode of action. Coagulation proceeded normally for the first minutes, and inhibition increased as equilibrium binding was established. It is suggested that association of fVIIa(i) with TF in a collision dependent reaction gives equal access of inhibitor and enzyme to TF. Assembly was not influenced by the higher affinity and slower dissociation of the dimer. As a result, anticoagulation was delayed until the reaction reached equilibrium. Properties of different dissociation experiments suggested that dissociation of fVIIai from TF occurred by a two-step mechanism. The first step was separation of TF-fVIIa(i) while both proteins remained bound to the membrane, and the second step was dissociation of the fVIIa(i) from the membrane. These results suggest novel actions of fVIIai that distinguish it from most of the anticoagulants that block later steps of the coagulation cascade. PMID- 15835922 TI - Probing the catalytically essential residues of the alpha-L-fucosidase from the hyperthermophilic archaeon Sulfolobus solfataricus. AB - Retaining glycosidases promote the hydrolysis of the substrate by following a double-displacement mechanism involving a covalent intermediate. The catalytic residues are a general acid/base catalyst and the nucleophile. Experimental identification of these residues in a specific glycosidase allows for the assigning of the corresponding residues in all of the other enzymes belonging to the same family. By means of sequence alignment, mutagenesis, and detailed kinetic studies of the alpha-fucosidase from Sulfolobus solfataricus (Ssalpha fuc) (family 29), we show here that the residues, invariant in this family, have the function inferred from the analysis of the 3D structure of the enzyme from Thermotoga maritima (Tmalpha-fuc). These include in Ssalpha-fuc the substrate binding residues His46 and His123 and the nucleophile of the reaction, previously described. The acid/base catalyst could be assigned less easily. The k(cat) of the Ssalpha-fucGlu292Gly mutant, corresponding to the acid/base catalyst of Tmalpha-fuc, is reduced by 154-fold but could not be chemically rescued. Instead, the Ssalpha-fucGlu58Gly mutant revealed a 4000-fold reduction of k(cat)/K(M) if compared to the wild-type and showed the rescue of the k(cat) by sodium azide at wild-type levels. Thus, our data suggest that a catalytic triad, namely, Glu58, Glu292, and Asp242, is involved in catalysis. Glu58 and Glu292 cooperate in the role of acid/base catalyst, while Asp242 is the nucleophile of the reaction. Our data suggest that in glycosidase family 29 alpha-fucosidases promoting the retaining mechanism with slightly different catalytic machineries coexist. PMID- 15835923 TI - The relationship between the branch-forming glycosyltransferases and cell surface sugar chain structures. AB - Many recombinant proteins developed or under development for clinical use are glycoproteins, and trials aimed at improving their bioactivity or pharmacokinetics in vivo by altering specific glycan structures are ongoing. For pharmaceuticals of glycoproteins, it is important to characterize and, if possible, control the glycosylation profile. However, the mechanism responsible for the regulation of sugar chain structures found on naturally occurring glycoproteins is still unclear. To clarify the relationship between glycosyltransferases and sugar chain branch structure, we estimated six glycosyltransferases' activities (N-acetylglucosaminyltransferase (GlcNAcTase)-I, -II, -III, -IV, -V, and beta-1,4-galactosyltransferase (GalT)) which control the branch formation on asparagine (Asn)-linked sugar chains in 18 human cancer cell lines derived from several tissues. To visualize the balance of glycosyltransferase activity associated with each cell line, we expressed the relative glycosyltransferase activity in comparison to the average activity among the cell lines. These cell lines were classified into five groups according to their relative glycosyltransferase balance and were termed GlcNAcTase-I/-II, GlcNAcTase-III, GlcNAcTase-IV, GlcNAcTase-V, and GalT. We also characterized the structures of Asn-linked sugar chains on the cell surface of representative cell lines of each group. The branching structure of cell surface sugar chains roughly corresponded to the glycosyltransferase balance. This finding suggests that, for the sugar chain structure remodeling of glycoproteins, attention should be focused on the glycosyltransferase balance of host cells before introducing exogenous glycosyltransferases or down-regulating the activity of intrinsic glycosyltransferases. PMID- 15835924 TI - Brain arachidonic acid incorporation is decreased in heart fatty acid binding protein gene-ablated mice. AB - Heart fatty acid binding protein (H-FABP) is expressed in neurons, but its role in brain fatty acid incorporation and metabolism is poorly defined. We examined the effect of H-FABP gene ablation on brain incorporation of arachidonic ([1 (14)C]20:4n-6) or palmitic ([1-(14)C]16:0) acid in vivo. Analysis of brain mRNA confirmed gene ablation and demonstrated no compensatory changes in the levels of other FABP mRNA in the gene-ablated mice. In brains from H-FABP gene-ablated mice, the incorporation coefficient for [1-(14)C]20:4n-6 was reduced 24%, while that for [1-(14)C]16:0 was unaffected. Within the organic and aqueous fractions, significantly more [1-(14)C]20:4n-6 was distributed into the aqueous fraction, suggesting a disruption in the metabolic targeting of 20:4n-6 in these mice. There was less incorporation of [1-(14)C]20:4n-6 into total phospholipids and a marked reduction (51%) in the level of incorporation into the choline glycerophospholipids (ChoGpl). Because FABP can influence steady-state lipid mass, brain individual lipid masses were measured. The brain total phospholipid mass was reduced 17% by gene ablation, ascribed to a 27% and 32% reduction in the masses of ChoGpl and sphingomyelin, respectively. Plasmalogen subclass masses were also reduced, suggesting that H-FABP may augment brain plasmalogen synthesis. In gene-ablated mice, the phosphatidylinositol 20:4n-6 level was reduced 25%, while the proportion of total n-6 fatty acids was reduced in the major phospholipid classes. Thus, these results demonstrate for the first time that H-FABP expression influences brain 20:4n-6 uptake and trafficking as well as steady-state brain lipid levels. PMID- 15835925 TI - Phosphatidylinositol 4,5-bisphosphate regulates activation-induced platelet microparticle formation. AB - While the role of the cytoskeleton in microparticle formation is well-described, the role of membrane phospholipids in regulating this process is poorly defined. PIP(2) binds many cytoskeletal proteins and may oppose microparticle formation through associations with these proteins. To determine whether PIP(2) effects microparticle formation, PIP(2) was incorporated into platelet membranes prior to activation-induced microparticle formation. Incorporation of PIP(2) into platelet membranes inhibited activation-induced microparticle formation by >or=90%. Inhibition was dose-dependent with an IC(50) of 12-18 microM. A permeabilized platelet system was next used to assess the effect of modulation of endogenous PIP(2) levels on microparticle formation. Infusion of type IIbeta PIP kinase into permeabilized platelets inhibited microparticle formation by 75 +/- 8%. In contrast, incubation of permeabilized platelets with PI-specific phospholipase C augmented microparticle formation by greater than 3-fold. Evaluation of PIP kinases following platelet activation demonstrated that they were lost from platelets in a calpain-dependent manner during microparticle formation. Purified mu-calpain cleaved recombinant type IIbeta PIP kinase and inhibited its ability to phosphorylate PI(5)P. In permeabilized platelets, incubation of purified mu calpain reduced PIP(2) levels, while exposure to calpeptin increased PIP(2) levels. Calpain has previously been implicated in platelet microparticle formation. These studies show that calpain may help limit PIP(2) formation following platelet activation and that PIP(2) content is an important determinant of platelet microparticle formation. PMID- 15835926 TI - Structure-reactivity studies of serum paraoxonase PON1 suggest that its native activity is lactonase. AB - PON1 is the best-studied member of a family of enzymes called serum paraoxonases, or PONs, identified in mammals (including humans) and other vertebrates as well as in invertebrates. PONs exhibit a range of important activities, including drug metabolism and detoxification of organophosphates such as nerve agents. PON1 resides on HDL (the "good cholesterol") and is also involved in the prevention of atherosclerosis. Despite this wealth of activities, the identity of PON1's native substrate, namely, the substrate for which this enzyme and other enzymes from the PON family evolved, remains unknown. To elucidate the substrate preference and other details of PON1 mechanism of catalysis, structure-activity studies were performed with three groups of substrates that are known to be hydrolyzed by PON1: phosphotriesters, esters, and lactones. We found that the hydrolysis of aryl esters is governed primarily by steric factors and not the pK(a) of the leaving group. The rates of hydrolysis of aliphatic esters are much slower and show a similar dependence on the pK(a) of the leaving group to that of the nonenzymatic reactions in solution, while the aryl phosphotriesters show much higher dependence than the respective nonenzymatic reaction. PON1-catalyzed lactone hydrolysis shows almost no dependence on the pK(a) of the leaving group, and unlike all other substrates, lactones seem to differ in their K(M) rather than k(cat) values. These, and the relatively high rates measured with several lactone substrates (k(cat)/K(M) approximately 10(6) M(-)(1) s(-)(1)) imply that PON1 is in fact a lactonase. PMID- 15835927 TI - Self-assembled magnetic nanowires made irreversible by polymer bridging. AB - In this letter, we investigate the mechanism of formation of a recently discovered new type of colloid, irreversible flexible chains of magnetic particles. The chain formation mechanism is based on magnetically induced bridging by adsorbed polymers, and we investigate here the associated phase diagrams, considering both thermodynamic and kinetic aspects. This phase diagram is the consequence of a balance between entropic repulsion between polymer layers at the particles surfaces, depletion forces pushing the particles together, and a short-range attractive force developing when polymers can bridge two particles. We end up with a very simple protocol allowing the formation of long, extremely regular chains, which can find numerous applications in chemistry and biology. The perspectives for the development of a new field of "macrocolloidal chemistry" are discussed. PMID- 15835928 TI - Carbon dioxide emulsion assisted loading of polymer microspheres toward sustained release materials. AB - An organic solvent-free method for encapsulating progesterone at high loadings within micron-sized inert latex polymer beads is reported. This approach makes use of a polymeric surfactant to emulsify carbon dioxide into an aqueous latex suspension. In this way, preformed approximately 4 microm polystyrene (PS) microparticles surface-grafted with poly(N-vinylpyrrolidone) (PVP) were plasticized and swollen followed by rapid partitioning of progesterone into the polymer matrix. The as-prepared polystyrene beads incorporated over 10% progesterone by weight while maintaining their initial size and morphological uniformity. Dissolution experiments were also carried out to obtain the release profile of progesterone entrapped within the PVP/PS particles. PMID- 15835929 TI - The internal dynamic modes of charged self-assembled peptide fibrils. AB - Photon correlation spectroscopy is used to study the internal dynamics of self assembled charged peptide fibrils. Short neutral and charged polymeric aggregates have diffusive modes due to whole macromolecular motion. For long semiflexible fibrils the logarithm of the intermediate scattering function follows a q(2)t(3/4) scaling at long times consistent with a Kratky-Porod free energy and preaveraged Oseen hydrodynamics. Persistence lengths on the order of micrometers are calculated for the peptide fibrils consistent with estimates from the liquid crystalline phase behavior. Fibril diameters (5-35 nm) calculated from the initial decay of the correlation functions are in agreement with transmission electron microscopy measurements. PMID- 15835930 TI - Continuous fabrication of biocatalyst immobilized microparticles using photopolymerization and immiscible liquids in microfluidic systems. AB - We report a novel technique for manufacturing polymeric microparticles containing biocatalysts by the behavior of immiscible liquids in microfluidic systems and in situ photopolymerization. The approach utilizes a UV-polymerizable hydrogel/enzyme solution and an immiscible oil solution. The oil and hydrogel solutions form emulsions in pressure-driven flow in microchannels at high values of the dimensionless capillary number (Ca). The resultant hydrogel droplets are then polymerized in situ via exposure to 365 nm UV light. This technique allows for the generation of monodisperse particles whose size can be controlled by the regulation of flow rates. In addition, both manufacturing microparticles and immobilizing biocatalysts can be performed simultaneously and continuously. PMID- 15835931 TI - Water induced hydrophobic surface. AB - A polyurethane coating is described that has hydrophilic wetting behavior when dry and hydrophobic when wet. A difference of approximately 25 degrees in advancing contact angles for dry (83 degrees ) and wet (108 degrees ) states is found by sessile drop and dynamic methods. The term "contraphilic" is suggested for this reversible change opposite customary amphiphilic behavior. Contraphilic behavior results from a soft block containing semifluorinated and 5,5 dimethyhydantoin segmers. Amide inter/intramolecular hydrogen bonding is proposed for the hydrophilic (dry) state, while surface-confined, amide-water hydrogen bonding "releases"semifluorinated groups, giving the hydrophobic state. Water induced hydrophobic surfaces may lead to applications for easily switched wetting, such as in microfluidics. PMID- 15835932 TI - Large-scale synthesis of high-quality ultralong copper nanowires. AB - The present difficulties in synthesis of one-dimensional copper are short length, nonlinear morphology, polydispersivity, poor crystallinity, low yield, and process complexity. In this work, we demonstrate that high-quality ultralong copper nanowires (90-120 nm in diameter, 40-50 microm in length; aspect ratio >350-450) can be synthesized in large scale with a facile aqueous reduction route at low cost. The prepared copper nanowires can also be used as starting solid precursor for fabrication of polycrystalline oxide nanotubes via direct oxidation in air. PMID- 15835933 TI - Ultralow interfacial tension for enhanced oil recovery at very low surfactant concentrations. AB - The interfacial tension (IFT) between alkanes and several individual surfactants and their mixtures has been investigated, using three kinds of alkyl hydrocarbons: decane, dodecane, and tetradecane. For individual and mixed surfactant systems, critical micelle concentrations and areas per molecule at the hydrocarbon-aqueous solution interface were calculated; for the mixed surfactant systems, betasigma(LL), the molecular interaction parameter at the hydrocarbon aqueous solution interface, and beta(M), the molecular interaction parameter in mixed micelle formation, were calculated. It was found that IFT in the 10(-3) mN/m (ultralow) range can be obtained at surfactant concentrations below 0.05 wt % and even at concentrations below 0.01 wt %, when mixtures of certain surfactants are used at the proper ratio. Surfactants with branched-chain alkyl groups show a much better IFT reduction effectiveness than those with straight chain alkyl groups. Contrary to what has been observed at the air-aqueous solution surface, mixtures of two homologues with two hydrophobic groups show significant molecular interactions, with both betasigma(LL) and beta(M) having negative values in the 4-5 range in some cases, with the betasigma(LL) value more negative than beta(M). The relationship between micellar shape and ultralow IFT was investigated by calculating the critical packing parameter of the surfactants. It was found that ultralow IFT between the surfactant mixtures and the three hydrocarbons investigated could reach ultralow (<10(-2) mN/m) values when the critical packing parameter is very close to 1. PMID- 15835934 TI - Solvation dynamics in aqueous anionic and cationic micelle solutions: sodium alkyl sulfate and alkyltrimethylammonium bromide. AB - Solvation dynamics of the fluorescence probe, coumarin 102, in anionic surfactant, sodium alkyl sulfate (C(n)H(2n+1)SO(4)Na; n = 8, 10, 12, and 14), and cationic surfactant, alkyltrimethylammonium bromide (C(n)H(2n+1)N(CH(3))(3)Br; n = 10, 12, 14, and 16), micelle solutions have been investigated by a picosecond streak camera system. The solvation dynamics in the time range of 10(-10)-10(-8) s is characterized by a biexponential function. The faster solvation time constants are about 110-160 ps for both anionic and cationic micelle solutions, and the slower solvation time constants for sodium alkyl sulfate and alkyltrimethylammonium bromide micelle solutions are about 1.2-2.6 ns and 450-740 ps, respectively. Both the faster and the slower solvation times become slower with longer alkyl chain surfactant micelles. The alkyl-chain-length dependence of the solvation dynamics in both sodium alkyl sulfate and alkyltrimethylammonium bromide micelles can be attributed to the variation of the micellar surface density of the polar headgroup by the change of the alkyl chain length. The slower solvation time constants of sodium alkyl sulfate micelle solutions are about 3.5 times slower than those of alkyltrimethylammonium bromide micelle solutions for the same alkyl-chain-length surfactants. The interaction energies of the geometry optimized mimic clusters (H(2)O-C(2)H(5)SO(4)(-) and H(2)O C(2)H(5)N(CH(3))(3)(+)) have been estimated by the density functional theory calculations to understand the interaction strengths between water and alkyl sulfate and alkyltrimethylammonium headgroups. The difference of the slower solvation time constants between sodium alkyl sulfate and alkyltrimethylammonium bromide micelle solutions arises likely from their different specific interactions. PMID- 15835935 TI - Flocculation with poly(ethylene oxide)/tyrosine-rich polypeptide complexes. AB - New insights into the mechanism for the flocculation of aqueous colloids by the sequential addition of a water-borne phenolic polymer, called cofactor, followed by very high molecular poly(ethylene oxide) (PEO) are presented. It is proposed that PEO/cofactor complexes form in the aqueous phase and adsorb onto the surfaces of the target colloidal particles. Flocculation will occur if PEO/cofactor complex on one particle will bind to adsorbed complex on a second particle; i.e., if the complexes are sticky. The proposed mechanism was illustrated by flocculation experiments with precipitated calcium carbonate, very high molecular weight PEO, and a polypeptide cofactor called PEY1 which was a 1:1 random copolymer of l-glycine and l-tyrosine. Independent measurements of the PEO/PEY1 complex properties, in the absence of calcium carbonate, were used to support the mechanism. In order for PEO/PEY1 complexes to be sticky, they must simultaneously have unbound PEY1 and polymer segments. With time the complexes deactivate (i.e., lose their stickiness) by a reconfiguration process which results in elimination of either unbound PEY1 or PEO segments. PMID- 15835936 TI - Single microgel particle studies demonstrate the influence of hydrophobic interactions between charged micelles and oppositely charged polyions. AB - The binding of two cationic surfactants, dodecyltrimethylammonium bromide (DoTAB) and N-(1,1,2,2-tetrahydroperfluorodecanyl)pyridinium bromide (HFDePB), to covalently cross-linked sodium poly(styrenesulfonate) (PSS) microgels has been investigated by means of micromanipulator-assisted time-resolved light microscopy on single gels. It is demonstrated that repeated measurements on the same microgel under conditions of controlled liquid flow give highly reproducible results. The two surfactants are found to behave very differently with respect to degree of swelling, surfactant distribution in the gels, both during shrinking and at equilibrium, and kinetics of volume changes induced by them. The main difference is attributed to the presence of a hydrophobic interaction between PSS and the DoTAB micelles, absent in the case of HFDePB. Kinetic shrinking curves are recorded and analyzed using a model for steady-state transport of surfactant between the solution and the gels. Aggregation numbers for DoTAB in PSS solutions obtained from fluorescence quenching measurements are presented. A strong dependence on the surfactant-to-polyion concentration ratio is observed. Relations between surfactant binding isotherms, phase diagrams for linear polyelectrolyte/surfactant/water systems, and the binding to gels are discussed. PMID- 15835937 TI - Use of a mesoporous material for organic synthesis. AB - A common problem in synthetic organic chemistry is attaining proper contact between lipophilic organic compounds and inorganic salts. Various strategies, for example, phase transfer catalysis (Starks, C. M.; Liotta, C. L.; Halpern, M. Phase Transfer Catalysis: Fundamentals, Applications and Industrial Perspectives; Chapman & Hall: New York, 1994) or use of a microheterogeneous medium such as a microemulsion (Hager, M.; Currie, F.; Holmberg, K. Organic Reactions in Microemulsions. In Colloid Chemistry II; Antonietti, M., Ed.; Topics in Current Chemistry 227; Springer-Verlag: Heidelberg, 2003; p 53) have been worked out to tackle the issue. Here, we report that mesoporous solid materials made from surfactant self-assembly can be used as medium for such reactions. The material is made from silica, and the pore size is large, relatively uniform, and can be controlled with a high degree of precision by the choice of surfactant that is being used as template (Palmqvist, A. E. C. Curr. Opin. Colloid Interface Sci. 2003, 8, 145). The pores are hydrophilic and are filled with an aqueous solution containing the inorganic salt. The porous material is dispersed in the lipophilic organic substrate, that is, 4-tert-butylbenzyl bromide, or in a hydrocarbon solution of this substrate. The reaction occurs at the hydrophilic/lipophilic interface, and, because the interface is large, the reaction is fast. A considerable advantage with this new reaction medium is that the workup procedure is extremely facile. After the reaction is completed, the solid is simply removed by filtering or centrifugation. PMID- 15835938 TI - Charge limits on droplets during evaporation. AB - We have examined charge stability limits of single evaporating microdroplets that were suspended in an electrodynamic balance. A high precision light scattering technique based on optical resonances was used to determine the size and the size change of a droplet at a charge instability induced breakup. The charge level and the charge loss at the breakup were obtained from the dc voltages required to balance the droplet prior to and following the breakup. The results on droplets of diethyl phthalate (DEP), diethylene glycol (DEG), triethylene glycol (TEG), and hexadecane show that breakups due to the charge instability occur at the Rayleigh charge limit. The observed charge losses during breakups range from about 15.3% for hexadecane droplets to about 41.1% for TEG droplets. Hexadecane droplets lose about 1.5% of their mass, while DEP droplets, about 2.3%. Within the detectable limit of 0.03%, no mass losses were observed during breakups of DEG and TEG droplets. The observation of extremely low mass losses that accompany high charge losses from DEG and TEG droplets suggests that the process of breakups of DEG and TEG droplets is distinct from that of DEP and hexadecane droplets. An analysis of the results indicates that breakups of DEP and hexadecane droplets result in the formation of a few large progeny droplets, while TEG and DEG droplets produce thousands of fine progeny droplets. PMID- 15835939 TI - 31P and 1h NMR as probes of domain alignment in a rigid crystalline surfactant mesophase. AB - A viscous reverse hexagonal surfactant mesophase containing bis(2-ethylhexyl) sodium sulfosuccinate (AOT) and alpha-phosphatidylcholine (lecithin), with comparable volume fractions of isooctane and water, was characterized by Fourier transform (31)P and (1)H NMR spectroscopy. Shear alignment was reflected through both (31)P NMR and (1)H NMR spectra. A complicated (31)P spectrum was observed as a result of superposition of chemical shifts according to the distribution of crystalline domains prior to shear. The initially disordered samples with polydomain structures become macroscopically aligned after Couette shear. (31)P NMR chemical shift anisotropy characteristics are used to elucidate orientation of the hexagonal phase. Interestingly, (1)H NMR spectra exhibit spectral changes upon shear alignment closely corresponding with that of (31)P NMR spectra. These observations complement the findings of mesophase alignment obtained using SANS and imply that (31)P and (1)H NMR spectroscopy can be used as probes to define microstructure and monitor orientation changes in this binary surfactant system. This is especially beneficial if these mesophases are used as templates for materials synthesis. PMID- 15835940 TI - Low molecular weight organogels based on long-chain carbamates. AB - Thermoreversible organogels were prepared from carbamates with alkyl side chains of different lengths. Gelation was possible only up to an alkyl side chain length of 12 carbons, beyond which crystallization occurs, due to the dominant van der Waals interaction between the alkyl chains. This is in contrast to other alkane based organogels, in which gelating efficiency increased with the length of the alkane chain (see Abdallah, D. J.; Weiss, R. G. Adv. Mater. 2000, 12, 1237). The critical concentration for gelation decreases drastically with an increase in the side chain length. Xerogels of these show birefringent fibers with uniform cross section and unlimited growth in one direction. The extent of this unlimited growth is affected by the length of the alkyl side chain in the carbamate, which finally ceases the gel formation ability of the carbamate. Cryogenic scanning electron microscopy images of the gels are similar to those of xerogels. From X ray diffraction of the fibers, we propose that the growth direction is along the plane of hydrogen bonds between the carbamate molecules. The thickness of the fibers depends on the length of the alkyl side chain. Morphological differences are seen between gels prepared by slow cooling and quenching of the solution. Thus, the morphology of the fibrous xerogels of the carbamates can be tailored for specific applications, by the choice of the alkyl side chain length and the rate of cooling the solution. PMID- 15835941 TI - Synthesis and characterization of shell cross-linked micelles with hydroxy functional coronas: a pragmatic alternative to dendrimers? AB - Shell cross-linked (SCL) micelles with hydroxy-functional coronas have been constructed in aqueous solution by exploiting the micellar self-assembly behavior of a new thermoresponsive ABC triblock copolymer. This copolymer was prepared via atom transfer radical polymerization in a convenient one-pot synthesis and comprised a thermoresponsive core-forming poly(propylene oxide) (PPO) block, a cross-linkable central poly(2-(dimethylamino)ethyl methacrylate) (DMA) block, and a hydroxy-functional outer block based on poly(glycerol monomethacrylate) (GMA). DMF GPC analysis confirmed a unimodal molecular weight distribution for the PPO PDMA-PGMA triblock copolymer precursor, with an M(n) of 12 100 and a polydispersity of approximately 1.26. This copolymer dissolved molecularly in aqueous solution at 5 degrees C but formed micelles with hydroxy-functional coronas above a critical micelle temperature of around 12 degrees C, which corresponded closely to the cloud point of the PPO macroinitiator. Cross-linking of the DMA residues using 1,2-bis(2-iodoethoxy)ethane produced SCL micelles that remained intact at 5 degrees C, i.e., below the cloud point of the core-forming PPO block. Dynamic light scattering studies confirmed that the SCL micelle diameter could be varied depending on the temperature employed for cross-linking: smaller, more compact SCL micelles were formed at higher temperatures, as expected. Since cross-linking involved quaternization of the DMA residues, the SCL micelles acquired cationic surface charge as judged by aqueous electrophoresis studies. These cationic SCL micelles were adsorbed onto near monodisperse anionic silica sols, which were used as a model colloidal substrate. Thermogravimetric analyses indicated a SCL micelle mass loading of 2.5-4.4%, depending on the silica sol diameter and the initial micelle concentration. Aqueous electrophoresis measurements confirmed that surface charge reversal occurred after adsorption of the SCL micelles, and scanning electron microscopy studies revealed a uniform coating of SCL micelles on the silica particles. PMID- 15835942 TI - Comparative studies on the micellization of sodium bis(4-phenylbutyl) sulfosuccinate and sodium bis(2-ethylhexyl) sulfosuccinate and their interaction with hydrophobically modified poly(acrylamide). AB - The micellization process of sodium bis(4-phenylbutyl) sulfosuccinate (SBPBS) has been studied compared to that of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) by surface tension, steady-state fluorescence, microcalorimetry, dynamic light scattering (DLS), and transmission electron microscopy (TEM) measurements. Meanwhile, the interaction of these two surfactants with hydrophobically modified poly(acrylamide) (HMPAM) was investigated. The results show that the surface tension at the critical micelle concentration (cmc) of SBPBS and the micropolarity probed by pyrene in SBPBS aggregates are both larger than those of AOT. The enthalpy change of micellization (DeltaH(mic)) of AOT is endothermic, while it is exothermic for SBPBS. Strong pi-pi interaction among the adjacent phenyl groups of SBPBS molecules is likely the cause for the above properties of SBPBS. Moreover, vesicles are observed for AOT and SBPBS by DLS and TEM, especially for AOT, whose micelle-vesicle transition has been first confirmed by its calorimetric curve. In the surfactant-HMPAM systems, the critical aggregation concentration (cac), the saturation concentration of aggregation (C(2)), and the thermodynamic parameters of binding have also been determined. The conclusion may be drawn that the binding strength of SBPBS onto HMPAM is stronger than that of AOT. PMID- 15835943 TI - Nonaqueous suspensions of laponite and montmorillonite. AB - It is shown how stable, nonaqueous suspensions of fully exfoliated smectite clays such as Laponite and montmorillonite can be obtained. Suspensions in toluene and in a branched aliphatic solvent (polydecene) were characterized using elemental analysis, rheometry, and small-angle X-ray scattering. For Laponite, stable suspensions were obtained by adsorbing a dichain poly(isobutylene) based stabilizer to the particles. This approach did not result in full exfoliation for montmorillonite particles, possibly because the stabilizer was able to connect individual clay sheets at the edges during the treatment process. Instead, a quaternary ammonium surfactant, dimethyl dioctadecylammonium bromide (DODAB), was first adsorbed to the clay. Subsequently, adsorption of the poly(isobutylene) based stabilizer to this pretreated clay resulted in fully exfoliated suspensions of montmorillonite. Suspensions of clay particles treated only with DODAB have been studied by several authors before and are included in this study for comparison. A detailed discussion of the suspension behavior of the different clay samples is given. The suspension routes presented here may enable further studies of the structure and flow behavior of suspensions of thin, flexible clay platelets as a function of aspect ratio. PMID- 15835944 TI - Experimental and computational growth morphology of two polymorphs of a yellow isoxazolone dye. AB - We report on the crystal structures and the experimentally found and the computationally predicted growth morphologies of two polymorphs of a yellow isoxazolone dye. The stable polymorph I has a blocklike habit, and the metastable polymorph II grows as fine needles, nucleating only by heterogeneous or contact nucleation. The habits of both polymorphs depend on the supersaturation during growth. The experimental observations are compared with predictions of the attachment energy model and kinetic Monte Carlo lattice simulations in which the growth is modeled as an "atomistic process", governed by surface energetics. These Monte Carlo simulations correctly predict the shape and the dependence on supersaturation of the crystal morphology for both polymorphs: for polymorph I, a strong dependence on supersaturation is found from the simulations. For polymorph II, the order of morphological importance is reproduced correctly, as well as the needlelike morphology. PMID- 15835945 TI - Adsorption of acicular particles at liquid-fluid interfaces and the influence of the line tension. AB - In this paper, the adsorption energy of an acicular (prolate and cylindrical) particle onto a liquid-fluid interface and the effect of the line tension are investigated. The results show that, without line tension, acicular particles always prefer to lie flat in the plane of the interface. However, line tension plays a significant role in determining the adsorption of an acicular particle. First, the line tension creates an energy barrier for the adsorption of particles onto an interface. The planar configuration has a larger energy barrier due to the longer contact line. Therefore, the particles prefer to enter the interface in a homeotropic configuration and then rearrange to a planar configuration or an oblique configuration with a small tilt angle. Second, for prolate particles, an energy maximum occurs at some tilt angles when the line tension is large. Therefore, once the prolate particle is adsorbed on the interface in a homeotropic configuration or with a larger tilt angle, it must conquer an energy barrier to rearrange to a planar configuration. For cylindrical particles, when the line tension is higher, the planar configuration will not be the most energy favorable configuration. The cylindrical particles prefer to stay in the interface with a small tilt angle. PMID- 15835946 TI - Modeling the properties of self-assembled monolayers terminated by carboxylic acids. AB - Self-assembled monolayers of long-chain carboxylic acids are often used as substrates to promote the growth of oriented crystals. Recent work has shown that the length of the chain (odd or even number of carbon atoms) determines whether oriented growth is observed. We use molecular dynamics simulations to investigate whether the configuration of the headgroups is significantly different in the two cases. We conclude that there are differences between odd- and even-length chains, even at 300 K and in the presence of water for some packings of the monolayer. We discuss whether these differences are large enough to account for the different behavior. PMID- 15835947 TI - Quantitative SAXS analysis of oriented 2D hexagonal cylindrical silica mesostructures in thin films obtained from nonionic surfactants. AB - Oriented mesostructured surfactant-silica nanocomposite thin films with a 2D hexagonal mesostructure of cylindrical micelles were prepared by evaporation induced self-assembly using two different nonionic Brij surfactants and studied by small-angle X-ray scattering in symmetric reflection (SRSAXS) and grazing incidence (GISAXS) geometries. A novel SRSAXS evaluation approach was applied that allowed a good fitting of the SRSAXS data over almost the whole range of scattering vectors. Aside from the cylinder radius and the lattice parameter, the approach provided accurate values for the polydispersity of the micelles, lattice distortions, and preferred orientation. These analyses revealed a significant rise of the micelle radius and accordingly the lattice parameter upon an increase in the ratio surfactant/SiO(2), attributable to a decrease in the solubilization of the poly(ethylene oxide) (PEO) chains by water, in agreement with Monte Carlo simulations. Furthermore, the SRSAXS analysis was successfully applied to the corresponding mesoporous films for the determination of pore sizes. PMID- 15835948 TI - Experimental study of entrainment and drainage flows in microscale soap films. AB - The thickness of freely suspended surfactant films during vertical withdrawal and drainage is investigated using laser reflectivity. The withdrawal process conducted at capillary numbers below 10(-3) generates initial film thicknesses in the micrometer range; subsequent thinning is predominantly impelled by capillary and not gravitational forces. Under these conditions, our results show that film thinning above and below the critical micelle concentration (cmc) is well approximated by a power law function in time whose exponents, which range from 0.9 to -1.8, are inconsistent with current descriptions of capillary-viscous drainage in inextensible films which predict exponents close to -0.5. Correlations between the experimental fitting parameters illustrate important differences in film behavior across the cmc. In addition, normalization of the drainage data yields a collapse to a single functional form over 3 decades in time for a wide range of initial withdrawal rates. We demonstrate that modification of the interface boundary condition in current models to account for Marangoni stresses through an effective slip parameter yields values of the exponents and other key parameters in excellent agreement with experiment. This modification also successfully describes the withdrawal thickness below the cmc. PMID- 15835949 TI - Dibenzodioxin adsorption on inorganic materials. AB - Dibenzodioxin adsorption/desorption on solid surfaces is an important issue associated with the formation, adsorption, and emission of dioxins. Dibenzodioxin adsorption/desorption behaviors on inorganic materials (amorphous/mesoporous silica, metal oxides, and zeolites) were investigated using in situ FT-IR spectroscopy and thermogravimetric (TG) analysis. Desorption temperatures of adsorbed dibenzodioxin are very different for different kinds of inorganic materials: approximately 200 degrees C for amorphous/mesoporous silica, approximately 230 degrees C for metal oxides, and approximately 450 degrees C for NaY and mordenite zeolites. The adsorption of dibenzodioxin can be grouped into three categories according to the red shifts of the IR band at 1496 cm(-1) of the aromatic ring for the adsorbed dibenzodioxin: a shift of 6 cm(-1) for amorphous/mesoporous silica, a shift of 10 cm(-1) for metal oxides, and a shift of 14 cm(-1) for NaY and mordenite, suggesting that the IR shifts are proposed to associated with the strength of the interaction between adsorbed dibenzodioxin and the inorganic materials. It is proposed that the dibenzodioxin adsorption is mainly via the following three interactions: hydrogen bonding with the surface hydroxyl groups on amorphous/mesoporous silica, complexation with Lewis acid sites on metal oxides, and confinement effect of pores of mordenite and NaY with pore size close to the molecular size of dibenzodioxin. PMID- 15835950 TI - Drops down the hill: theoretical study of limiting contact angles and the hysteresis range on a tilted plate. AB - The limiting inclination angle (slip angle), for which a two-dimensional water drop may be at equilibrium on a chemically heterogeneous surface, is exactly calculated for a variety of cases. The main conclusion is that, in the cases studied, the contact angles at the upper and lower contact line do not always simultaneously equal the receding and advancing contact angles, respectively. On a hydrophobic surface, the lowest contact angle (at the upper contact line) tends to be approximately equal to the receding contact angle, while the highest contact angle (at the lower contact line) may be much lower than the advancing contact angle. For hydrophilic surfaces, the opposite is true. These conclusions imply that the hysteresis range cannot in general be measured by analyzing the shape of a drop on an inclined plane. Also, the limiting inclination angle cannot in general be calculated from the classical equation based only on the advancing and receding contact angles. PMID- 15835951 TI - TPD and FT-IRAS investigation of ethylene oxide (EtO) adsorption on a Au(211) stepped surface. AB - Adsorption of ethylene oxide, CH(2)CH(2)O (EtO), on a Au(211) stepped surface was studied by temperature programmed desorption (TPD) and Fourier transform infrared reflection-absorption spectroscopy (FT-IRAS). Ethylene oxide was completely reversibly adsorbed, and desorbed molecularly during TPD following adsorption on Au(211) at 85 K. EtO TPD peaks appeared at 115 K from the multilayer film and 140 and 170 K from the monolayer. Desorption at 140 K was attributed to EtO desorption from terrace sites, and that at 170 K to EtO desorption from step sites. Desorption activation energies and corresponding adsorption energies were estimated to be 8.4 and 10.3 kcal mol(-1), respectively. The EtO ring (C(2)O) deformation band appeared in IRAS at 865 cm(-1) for EtO in multilayer films and when adsorbed in the monolayer at terrace sites. The stronger chemisorption bonding of EtO at Au step sites slightly weakens the bonding within the molecule and causes a small red-shift of this band to 850 cm(-1) for adsorption at step sites. EtO presumably binds via the oxygen atom to the surface, and observation of the EtO-ring absorption band in IRAS establishes that the molecular ring plane of EtO adsorbed at step and terrace sites is nearly upright with respect to the crystal surface plane. PMID- 15835952 TI - Adsorption of aqueous metal ions on oxygen and nitrogen functionalized nanoporous activated carbons. AB - In this study, the adsorption characteristics of two series of oxygen and nitrogen functionalized activated carbons were investigated. These series were a low nitrogen content (approximately 1 wt % daf) carbon series derived from coconut shell and a high nitrogen content (approximately 8 wt % daf) carbon series derived from polyacrylonitrile. In both series, the oxygen contents were varied over the range approximately 2-22 wt % daf. The porous structures of the functionalized activated carbons were characterized using N(2) (77 K) and CO(2) (273 K) adsorption. Only minor changes in the porous structure were observed in both series. This allowed the effect of changes in functional group concentrations on metal ion adsorption to be studied without major influences due to differences in porous structure characteristics. The surface group characteristics were examined by Fourier transform infrared (FTIR) spectroscopy, acid/base titrations, and measurement of the point of zero charge (pH(PZC)). The adsorption of aqueous metal ion species, M(2+)(aq), on acidic oxygen functional group sites mainly involves an ion exchange mechanism. The ratios of protons displaced to the amount of M(2+)(aq) metal species adsorbed have a linear relationship for the carbons with pH(PZC) < or = 4.15. Hydrolysis of metal species in solution may affect the adsorption of metal ion species and displacement of protons. In the case of basic carbons, both protons and metal ions are adsorbed on the carbons. The complex nature of competitive adsorption between the proton and metal ion species and the amphoteric character of carbon surfaces are discussed in relation to the mechanism of adsorption. PMID- 15835953 TI - Measurement of the kinetics of photo-oxidation of self-assembled monolayers using friction force microscopy. AB - The advancement of molecular nanotechnology requires new tools for the characterization of surface chemical reactivity with nanometer spatial resolution. While spectroscopy on sub-100 nm length scales remains challenging, friction force microscopy (FFM) is a promising tool for the characterization of molecular materials, although to date it has been little used in studies of surface reactivity. Here we report the use of FFM to measure the kinetics of photo-oxidation of self-assembled monolayers (SAMs) of alkanethiols adsorbed on gold surfaces. Two alternative approaches (analysis of friction-load plots and the use of line sections through images of patterned materials) are compared and found to yield data in very good agreement, with rate constants being found to be in good agreement despite being carried out on different microscopes. The use of line-section analysis provides a convenient method for the quantification of the extent of reaction in nanometer-scale patterns created in SAMs by the novel approach of scanning near-field photolithography. PMID- 15835954 TI - Microchannel systems in titanium and silicon for structural and mechanical studies of aligned protein self-assemblies. AB - We report a technique for the alignment of self-assembled protein systems, such as F-actin bundles and microtubules, in a surface-modified titanium or silicon microfluidic device. Assembling filamentous protein systems in a confined geometry produces highly aligned samples for structural and mechanical studies. Biomolecular self-assembly can be investigated in a controlled fashion under different molecular concentration gradients and conditions along a channel length. We have shown that surface-modified devices produced via a high aspect ratio etch process in titanium and silicon can be used to confine and control such macromolecular assemblies and present examples of F-actin bundles and microtubules in this system. PMID- 15835955 TI - Kinetic modeling of the adsorption rate of a gaseous adsorbate on a granular adsorbent by ultra-rapid-scanning Fourier transform infrared spectrometry. AB - A novel method for determining the rates of adsorption of gaseous adsorbates on granular adsorbents in the millisecond time regime is described. Mixtures of the adsorbate and nitrogen are passed through a solenoid valve and 100 mg of the adsorbent for periods of (typically) 100 ms. The concentration of the adsorbate passing into a low-volume, long path length gas cell is measured with an ultra rapid-scanning Fourier transform infrared spectrometer capable of gathering 200 mid-infrared spectra per second with 6-cm(-1) resolution. The pressure of the gas entering the cell is measured simultaneously with a capacitance manometer. A dynamic mathematical model was developed to analyze and describe the results in terms of a Langmuir isotherm. The success of this approach is demonstrated by the estimation of the rate of adsorption of vapor-phase acetaldehyde on aminopropylsilylated granular silica gel. PMID- 15835956 TI - Kinetic modeling of aldehyde adsorption rates on bare and aminopropylsilyl modified silica gels by ultra-rapid-scanning fourier transform infrared spectrometry. AB - Enhancements of a model capable of precisely defining rates of adsorption in terms of a Langmuir isotherm and determined from infrared and pressure data collected in the millisecond time regime are described in detail. Rates of adsorption are determined for formaldehyde and acetaldehyde exposed to bare and aminopropylsilyl-derivatized silica gel. The model fits the collected data by varying the number of adsorption sites and the adsorption and desorption rate constants and determines the best fit based on the measured IR absorbance, partial pressure, exposure backpressure, number of adsorption sites, and time. The rate of adsorption onto irregular-shaped silica gel was faster than the rate onto the corresponding spherical silica gel. Formaldehyde was adsorbed more efficiently than acetaldehyde on APS silica gel than on bare silica gel, whereas acetaldehyde was adsorbed more efficiently than formaldehyde on underivatized silica, indicating the possibility that formaldehyde is adsorbed through the formation of a Schiff base. PMID- 15835957 TI - Comparative study of the adhesion, friction, and mechanical properties of CF3- and CH3-terminated alkanethiol monolayers. AB - We report the results of a direct comparison of the adhesion, friction, and mechanical properties between alkanethiol self-assembled monolayer films terminated by either CH(3) or CF(3) end groups using both interfacial force (IFM) and atomic force (AFM) microscopies. The purpose of this work is to gain insight into the detailed origins of the differing frictional behavior previously observed with AFM. The IFM results reveal an increased adhesive interaction for the CF(3)-terminated film due to the highly polar nature of the end groups. In agreement with earlier studies, the AFM results show two linear regions with differing frictional slopes for the CH(3)-terminated film but only a single slope for the CF(3)-terminated film. We contrast the differences between these techniques, approximately 100 times smaller tips for the AFM, and discuss the role of the mechanical properties, the increased adhesive interaction, and the amount of disorder present in the film in creating differences in frictional behavior between the two systems. We conclude that increased adhesion for the CF(3)-terminated film plays an important role in the observed differences in frictional behavior, while the differences between the two techniques can be traced to the different tip sizes and the consequent responses to the presence of disorder in the films. PMID- 15835958 TI - The adsorption of C4 unsaturated hydrocarbons on highly dehydrated silica. An IR spectroscopic and thermodynamic study. AB - The adsorptive interaction of 1-butyne and 1-butene with a highly dehydrated pyrogenic silica system has been studied to understand the thermodynamic behavior of the adsorption process by the application of the Langmuir model and of the Van't Hoff equation. In situ FTIR spectroscopy allowed the characterization of the adsorption phenomenon in terms of involved surface sites, gas-volumetric determinations yielded quantitative information relative to the adsorption process, and microcalorimetric results allowed the comparison between calculated and experimental data. K(eq) and Delta(ads)G degrees were obtained from Langmuir's model application; Delta(ads)H data were obtained from the Van't Hoff equation and by the isosteric heats method and were compared with experimental values. The virtual constancy of Delta(ads)H with equilibrium pressure and surface coverage (Langmuir model) allowed us to obtain the Delta(ads)H degrees values and, consequently, the Delta(ads)S degrees values for the systems of interest. PMID- 15835959 TI - Thin films of hydrophobically modified poly(N,N-dimethylacrylamide). AB - The behavior of a poly(N,N-dimethylacrylamide) hydrophobically modified by incorporating 0.33 mol % of a pyrenyl derivative, [4-(1-pyrenyl)butyl]amine hydrochloride (PY) and 3.56 mol % of dodecylamine (DO) has been studied at the air/water interface. Surface pressure-area isotherm measurements show that the film is initially anchored by the hydrophobic groups at the air-water interface with a pancake-like structure and, with increasing surface pressure, evolves to a quasi mushroom structure, finally reaching a brush configuration at high pressures. Monolayers of this polymer were transferred to silica substrates using the Langmuir-Blodgett (LB) technique at 5, 15, and 25 mN.m(-1). The properties of the LB films were studied by steady-state and time-resolved fluorescence as well as by atomic force microscopy. The results show that the aggregates formed at low pressures are disrupted by pressure increase, while the water-soluble poly(N,N dimethylacrylamide) becomes dissolved in the water subphase. PMID- 15835960 TI - Interactions of hydrophobically modified polyelectrolytes with nonionic surfactants. AB - Interactions of surfactants with hydrophobically modified polyelectrolytes in aqueous solutions are important in several applications such as detergency, cosmetics, food, and paints. Complexes formed in these systems raise some fundamental questions about the polymer-surfactant interactions that control their behavior. In this work, the interactions of a nonionic surfactant, penta ethyleneglycol mono n-dodecyl ether (C(12)EO(5)), with a hydrophobically modified anionic polymer, poly(maleic acid/octyl vinyl ether) (PMAOVE), in aqueous solutions were studied using surface tension, viscosity, electron paramagnetic resonance (EPR) spectroscopy, light scattering, and fluorescence spectroscopic techniques. When the nonionic surfactant C(12)EO(5) was added to aqueous solutions of the anionic polymer PMAOVE, it was incorporated into the hydrophobic nanodomains of PMAOVE far below the the critical micelle concentration (cmc) of the surfactant. Two inflection points were observed corresponding to the critical complexation concentration (formation of mixed micelles composed of C(12)EO(5) and the octyl chains of PMAOVE) and the saturation concentration (saturation of the polymer with C(12)EO(5) molecules). Above the saturation concentration, the coexistence of pure C(12)EO(5) micelles and mixed micelles of PMAOVE and C(12)EO(5) was observed. Such a coexistence of complexes has major implications in their performance in colloidal processes. PMID- 15835962 TI - Analysis of the microfluid flow in an evaporating sessile droplet. AB - The axisymmetric time-dependent flow field in an evaporating sessile droplet whose contact line is pinned is studied numerically and using an analytical lubrication theory with a zero-shear-stress boundary condition on the free surface of the droplet at low capillary and Reynolds numbers. A finite element algorithm is developed to solve simultaneously the vapor concentration and flow field in the droplet under conditions of slow evaporation. The finite element solution confirms the accuracy of the lubrication solution, especially when terms of higher order in the droplet flatness ratio (the ratio of droplet height to radius, h/R) are included in the lubrication theory to account more accurately for the singular flow near the contact line. PMID- 15835961 TI - Programmed hyperhelical supramolecular assembly of nickel phthalocyanine bearing enantiopure 1-(p-tolyl)ethylaminocarbonyl groups. AB - The present paper reports uniqueness of a simple, programmed design of disk shaped homochiral nickel phthalocyanine (Pc) molecules bearing four enantiomerically pure 1-(p-tolyl)ethylaminocarbonyl groups at their peripheral positions, (Pc-(R) and Pc-(S)), and their controlled self-organization into mesoscopic supramolecular helical fibers with a preferential handedness in solution and onto solid surfaces. A combination of four fundamental intermolecular interactions, including quadruple hydrogen bonding, pi-pi stacking, homochiral interactions of the enantiopure bulky aralkyl entities, and noncoordinating nature of nickel ion of the Pc molecules afforded a high thermal stability of the Pc self-assembly in chloroform (CHCl(3)), tetrahydrofuran, and o dichlorobenzene and onto hydrophilic mica and hydrophobic HOPG surfaces. A higher ordered helical self-assembly of Pc disks was observed in these solutions (approximately 200 Pc molecules), while the self-assembly was completely dissociated into monomeric species in N,N-dimethylformamide due to a loss of hydrogen-bonding interactions between Pc molecules. Supramolecular chirality in the hierarchical self-assembly of Pc molecules originated from the presence of (R)- or (S)-chiral centers in the peripheral tails, which rotate noncovalently linked molecular building blocks to effectively form the helical architectures. The helical Pc nanofibers dissolved in CHCl(3), estimated to be ca. 70 nm from peak molecular weight obtained by SEC analysis, acts as a building block for higher-order helical fibers (ca. 1 microm) at single molecular level on the solid surfaces, as demonstrated by the dynamic force mode atomic force microscopy. Regardless of hydrophilic and hydrophobic substrates, the interaction between these Pc molecules and the solid surfaces could not affect the morphology of helical assemblies, indicating a unique robustness of these assemblies. PMID- 15835963 TI - Analysis of the effects of Marangoni stresses on the microflow in an evaporating sessile droplet. AB - We study the effects of Marangoni stresses on the flow in an evaporating sessile droplet, by extending a lubrication analysis and a finite element solution of the flow field in a drying droplet, developed earlier. The temperature distribution within the droplet is obtained from a solution of Laplace's equation, where quasi steadiness and neglect of convection terms in the heat equation can be justified for small, slowly evaporating droplets. The evaporation flux and temperature profiles along the droplet surface are approximated by simple analytical forms and used as boundary conditions to obtain an axisymmetric analytical flow field from the lubrication theory for relatively flat droplets. A finite element algorithm is also developed to solve simultaneously the vapor concentration, and the thermal and flow fields in the droplet, which shows that the lubrication solution with the Marangoni stress is accurate for contact angles as high as 40 degrees. From our analysis, we find that surfactant contamination, at a surface concentration as small as 300 molecules/microm(2), can almost entirely suppress the Marangoni flow in the evaporating droplet. PMID- 15835964 TI - Morphosynthesis of strontianite nanowires using polyacrylate templates tethered onto self-assembled monolayers. AB - Strontianite nanowires have been synthesized on self-assembled monolayers (SAM) in the presence of polyacrylate templates. The morphology of this product exhibits characteristic differences from that of products obtained in the absence of polyacrylate. It is demonstrated that the template-induced crystallization process involves the interaction between the SAM surface, polyacrylate (a dissolved polyelectrolyte), and the cations/anions in solution. By the combination of these components, hierarchically ordered mineral hybrid structures are formed. PMID- 15835965 TI - Controlled crystallization of CaCO(3) on hyperbranched polyglycerol adsorbed to self-assembled monolayers. AB - The formation of biominerals by living organisms is governed by the cooperation of soluble and insoluble macromolecules with peculiar interfacial properties. To date, most of the studies on mineralization processes involve model systems that only account for the existence of one organic matrix and thus disregard the interaction between the soluble and insoluble organic components that is crucial for a better understanding of the processes taking place at the inorganic-organic interface. We have set up a model system composed of a matrix surface, namely, a self-assembled monolayer (SAM), and a soluble component, hyperbranched polyglycerol. The model mineral calcium carbonate displays diverse polymorphism. It could be demonstrated that the phase selection of calcium carbonate is controlled by the cooperative interaction of the SAM and hyperbranched polyglycerol of different molecular weights (M(n) = 500-6000 g/mol) adsorbed to the SAM. Our studies showed that hyperbranched polyglycerol is adsorbed to polar as well as to nonpolar SAMs. This effect can be related to its highly flexible structure and its amphiphilic character. The adsorption of hyperbranched polyglycerol to the SAMs with different surface polarities resulted in the formation of aragonite for alkyl-terminated SAMs and no phase selection for carboxylate-terminated SAMs. PMID- 15835966 TI - Energy transfer between chlorophyll derivatives in silica mesostructured films and photocurrent generation. AB - Layered silica/surfactant mesostructured thin films containing chlorophyllous pigments [C13(2)-demethoxycarbonyl-pheophytin b (pyroPheo b) or zinc C13(2) demethoxycarbonyl-chlorophyll b (Zn-pyroChl b)] have been prepared on an indium tin oxide (ITO) electrode grafted with a chlorophyll derivative possessing a triethoxysilyl group (copper C13(2)-demethoxycarbonyl-chlorophyllide a 3 triethoxysilyl propylamide, Cu-APTES-Chl a) to achieve effective light harvesting and successive photocurrent generation by the mesostructured films. The incorporation of pyroPheo b and Zn-pyroChl b in the mesostructured film resulted in 1.2- and 1.6-fold increases of the photocurrent density, respectively, as compared to the case of an antenna pigment-free film also grafted to a surface modified ITO electrode. The difference action spectra, between the electrodes with and without the antenna pigments, coincided well with the absorption spectra of the immobilized pigments. Because direct electron injection from the antenna pigments in the mesostructured films to the ITO electrode was scarcely observed, the energy transfer from the antenna pigments to Cu-APTES-Chl a plays an important role for the increase in photocurrent density. The usefulness of the mesostructured films as model systems is discussed in relation to the photosynthetic primary processes of higher plants. PMID- 15835967 TI - Thermal decomposition of generation-4 polyamidoamine dendrimer films: decomposition catalyzed by dendrimer-encapsulated Pt particles. AB - The thermal decomposition of hydroxyl-terminated generation-4 polyamidoamine dendrimer (G4OH) films deposited on Au surfaces has been compared with decomposition of the same dendrimer encapsulating an approximately 40-atom Pt particle (Pt-G4OH). Infrared absorption reflection spectroscopy studies showed that, when the films were heated in air to various temperatures up to 275 degrees C, the disappearance of the amide vibrational modes occurred at lower temperature for the Pt-G4OH film. Dendrimer decomposition was also investigated by thermogravimetric analysis (TGA) in both air and argon atmospheres. For the G4OH dendrimer, complete decomposition was achieved in air at 500 degrees C, while decomposition of the Pt-G4OH dendrimer was completed at 400 degrees C, leaving only platinum metal behind. In a nonoxidizing argon atmosphere, a greater fraction of the G4OH decomposed below 300 degrees C, but all of the dendrimer fragments were not removed until heating above 550 degrees C. In contrast, Pt G4OH decomposition in argon was similar to that in air, except that decomposition occurred at temperatures approximately 15 degrees C higher. Thermal decomposition of the dendrimer films on Au surfaces was also studied by temperature programmed desorption (TPD) and X-ray photoelectron spectroscopy (XPS) under ultrahigh vacuum conditions. Heating the G4OH films to 250 degrees C during the TPD experiment induced the desorption of large dendrimer fragments at 55, 72, 84, 97, 127, 146, and 261 amu. For the Pt-G4OH films, mass fragments above 98 amu were not observed at any temperature, but much greater intensities for H(2) desorption were detected compared to that of the G4OH film. XPS studies of the G4OH films demonstrated that significant bond breaking in the dendrimer did not occur until temperatures above 250 degrees C and heating to 450 degrees C caused dissociation of C=O, C-O, and C-N bonds. For the Pt-G4OH dendrimer films, carbon-oxygen and carbon-nitrogen bond scission was observed at room temperature, and further decomposition to atomic species occurred after heating to 450 degrees C. All of these results are consistent with the fact that the Pt particles inside the G4OH dendrimer catalyze thermal decomposition, allowing dendrimer decomposition to occur at lower temperatures. However, the Pt particles also catalyze bond scission within the dendrimer fragments so that decomposition of the dendrimer to gaseous hydrogen is the dominant reaction pathway compared to desorption of the larger dendrimer fragments observed in the absence of Pt particles. PMID- 15835968 TI - X-ray absorption spectroscopy to probe surface composition and surface deprotection in photoresist films. AB - Near-edge X-ray absorption fine structure spectroscopy (NEXAFS) is utilized to provide insight into surface chemical effects in model photoresist films. First, NEXAFS was used to examine the resist/air interface including surface segregation of a photoacid generator (PAG) and the extent of surface deprotection in the film. The concentration of PAG at the resist-air interface was higher than the bulk concentration, which led to a faster deprotection rate at that interface. Second, a NEXAFS depth profiling technique was utilized to probe for compositional gradients in model resist line edge regions. In the model line edge region, the surface composition profile for the developed line edge was dependent on the post exposure bake time. PMID- 15835969 TI - Adsorption behavior of surface chemically pure N-cycloalkylaldonamides at the air/water interface. AB - Equilibrium surface tension (sigma(e)) and electric surface potential (DeltaV(e)) versus concentration isotherms of the homologous series of N cycloalkylaldonamides synthesized from cycloalkylamines (from cyclopentyl- to cyclododecylamine) and D-glucono-1,5-lactone (c-C(n)GA) or D-glucoheptono-1,4 lactone (c-C(n)GHA) (c-n(C) = 5-12) were investigated at the air/water interface. The measurements were performed with aqueous, surface chemically pure surfactant solutions. Equilibrium surface tension vs concentration isotherms were evaluated to get the adsorption parameters, i.e., standard free energy of adsorption, DeltaG degrees (ads), saturation surface concentration, Gamma(infinity), minimum surface area demand per molecule adsorbed, A(min), and interaction parameter, H(s). Increasing the size of the cycloalkyl moiety leads to a significant increase of the minimum surface area demand per molecule adsorbed. This fact, together with a decrease of the intermolecular interaction parameter suggests that the introduction of a more bulky cycloalkyl ring (c-n(C) = 7 and 8) causes an attenuation of the hydrogen-bond network. This goes in line with the exceptional finding that the higher homologues revealed improved solubility in water. In addition, surface tension investigations suggest occurrence of a phase transition for the N-cyclooctylaldonamides at relatively small surface coverage. This observation is well supported by the surface potential measurements, for which the effect of possible changes in the molecules' surface orientation is even more pronounced. Moreover, the concentration intervals of N cyclooctylaldonamide in which the change in orientation is observed for either the surface tension or the surface potential isotherms are in very good agreement. PMID- 15835970 TI - In Situ dynamic monitoring of electrochemical oxidative adsorption and reductive desorption processes of a self-assembled monolayer of hexanethiol on a Au(111) surface in KOH ethanol solution by scanning tunneling microscopy. AB - Electrochemical oxidative formation and reductive desorption processes of a self assembled monolayer (SAM) of hexanethiol on a Au(111) surface in KOH ethanol solutions containing various concentrations of hexanethiol were investigated by in situ scanning tunneling microscopy in real time. The generation and disappearance of vacancy islands (VIs), corresponding to the formation and desorption of the SAM, respectively, were observed as anodic and cathodic current, respectively, flowed when the thiol concentration was higher than ca. 1 microM. When the VIs disappeared after the reductive desorption of the SAMs, the herringbone structure corresponding to the (radical3 x 23) structure of Au(111), was observed on the surface, indicating that a clean reconstructed surface was exposed even in the hexanethiol ethanol solution. During both oxidative adsorption and reductive desorption of the SAMs, the shape of the steps of the gold substrate changed drastically and the step lines became parallel to the 121 direction of the Au(111) surface, suggesting that gold atoms on the surface were extremely mobile during these processes. The coalescence of adjacent vacancy islands and growth of larger islands triangular in shape accompanied with the disappearance of nearby smaller islands were observed, confirming that the VIs grew according to the Ostward ripening model. PMID- 15835971 TI - Film formation from aqueous polyurethane dispersions of reactive hydrophobic and hydrophilic components; spectroscopic studies and Monte Carlo simulations. AB - Film formation of waterborne two-component polyurethanes is exceedingly complex due to the heterogeneous nature along with simultaneous progression of several parallel physicochemical processes which include water evaporation, cross-linking reactions, phase separation, and droplet coalescence, to name a few. While internal reflection infrared imaging (IRIRI) spectroscopy clearly facilitates analysis of chemical changes resulting from film formation, the complexity of processes leading to formation of specific surface/interfacial entities is a major experimental challenge. For this reason, we combined a spectrum of surface/interfacial analytical approaches including IRIRI, atomic force microscopy, and attenuated total reflectance Fourier transform infrared spectroscopy with Monte Carlo computer simulations to advance the limited knowledge of how temperature, stoichiometry, concentration levels, and reactivities of individual components affect the development of surface morphologies and compositional gradients across the film thickness. These studies show that in heterogeneous systems having both hydrophobic and hydrophilic components stratification of individual components to the film-air (F-A) interface is ultimately responsible for formation of rough surface topographies. These studies show that simultaneous stratification of hydrophobic components along with water evaporation to the F-A interface results in metastable interfacial layers, leading to surface dewetting. Subsequently, surface roughness is enhanced by higher concentrations of water in the cross-linking film. PMID- 15835972 TI - Development of a regenerable cell culture system that senses and releases dead cells. AB - We developed a rapidly regenerable cell culture system in which the cell culture substrate detects cell death and selectively releases the dead cells. This culture material was achieved by combining a detector that responds to the signal from the dead cells and an actuator to release the dead cells. Benzo-18-crown-6 acrylamide (BCAm) with a pendant crown ether receptor was used as the sensor to recognize cellular signals and N-isopropylacrylamide (NIPAM) was used as the actuator. This copolymer of NIPAM and BCAm can respond to potassium ions and change its nature from hydrophobic to hydrophilic at the culture temperature of 37 degrees C. Living cells concentrate potassium ion internally; when cells die, potassium ions are released. The polymer surface recognizes the potassium ions released from the dead cells, the NIPAM hydrates, and the dead cells are selectively detached. This in vitro culture system is a novel one in which artificial culture materials work cooperatively with cellular metabolism by responding to this signal from the cells, thereby realizing in vitro tissue regeneration partly mimicking the mechanisms of in vivo homeostasis. PMID- 15835973 TI - Surface enzyme kinetics for biopolymer microarrays: a combination of Langmuir and Michaelis-Menten concepts. AB - Real-time surface plasmon resonance (SPR) imaging measurements of surface enzymatic reactions on DNA microarrays are analyzed using a kinetics model that couples the contributions of both enzyme adsorption and surface enzyme reaction kinetics. For the case of a 1:1 binding of an enzyme molecule (E) to a surface immobilized substrate (S), the overall enzymatic reaction can be described in terms of classical Langmuir adsorption and Michaelis-Menten concepts and three rate constants: enzyme adsorption (k(a)), enzyme desorption (k(d)) and enzyme catalysis (k(cat)). In contrast to solution enzyme kinetics, the amount of enzyme in solution is in excess as compared to the amount of substrate on the surface. Moreover, the surface concentration of the intermediary enzyme-substrate complex (ES) is not constant with time, but goes to zero as the reaction is completed. However, kinetic simulations show that the fractional surface coverage of ES on the remaining unreacted sites does reach a steady-state value throughout the course of the surface reaction. This steady-state value approaches the Langmuir equilibrium value for cases where k(a)[E] >> k(cat). Experiments using the 3' --> 5' exodeoxyribonuclease activity of Exonuclease III on double-stranded DNA microarrays as a function of temperature and enzyme concentration are used to demonstrate how this model can be applied to quantitatively analyze the SPR imaging data. PMID- 15835974 TI - Study of Langmuir and Langmuir-Blodgett films of odorant-binding protein/amphiphile for odorant biosensors. AB - To make ultrathin films for the fabrication of artificial olfactory systems, odorant biosensors, we have investigated mixed Langmuir and Langmuir-Blodgett films of odorant-binding protein/amphiphile. Under optimized experimental conditions (phosphate buffer solution, pH 7.5, OBP-1F concentration of 4 mg L( 1), target pressure 35 mN m(-1)), the mixed monolayer at the air/water interface is very stable and has been efficiently transferred onto gold supports, which were previously functionalized by self-assembled monolayers (SAMs) with 1 octadecanethiol (ODT). Atomic force microscopy and electrochemical impedance spectroscopy were used to characterize mixed Langmuir-Blodgett (LB) films before and after contact with a specific odorant molecule, isoamyl acetate. AFM phase images show a higher contrast after contact with the odorant molecule due to the new structure of the OBP-1F/ODA LB film. Non-Faradaic electrochemical spectroscopy (EIS) is used to quantify the effect of the odorant based on the electrical properties of the OBP-1F/ODA LB film, as its resistance strongly decreases from 1.18 MOmega (before contact) to 25 kOmega (after contact). PMID- 15835975 TI - First preparation of biotinylated gradient polyethylene surface to bind photoactive caged streptavidin. AB - A gradient polyethylene (PE) surface was created through corona treatment, in which the corona power increased along the 5 cm length of the PE. The gradient surface was treated with polyethyleneimine and then biotin. Fluorescein conjugated streptavidin (SAV) caged within 5-carboxymethoxy-2-nitrobenzyl (CMNB) adsorbed onto the biotinylated gradient PE surface following molecular recognition principles. Photoirradiation decomposed the CMNB cage and allowed the fluorescein-conjugated SAV to fluoresce, the intensity of which increased gradually along the PE surface. PMID- 15835976 TI - Photoelectric performance of bacteria photosynthetic proteins entrapped on tailored mesoporous WO3-TiO2 films. AB - Novel three-dimensional wormlike mesoporous WO(3)-TiO(2) films with tailored pore size (approximately 7.1 nm) were applied to prepare the bio-photoelectrodes (Bio PEs) through direct entrapping the bacteria photosynthetic reaction center (RC) proteins. These mesoporous WO(3)-TiO(2) films exhibited unique characteristics in the specific loading of RC with high activity retained. Moreover, well-matched energy levels of WO(3)-TiO(2) and RC contributed to the photoelectric performance, especially in the red to near-infrared (NIR) region, of the derived Bio-PEs. Such strategy of manipulating the Bio-PEs based on well-designed mesoporous metal oxides and RC provides an alternative system to probe the photoinduced multiple-pathway electron transfer of photosensitive chromophores, which may open a new perspective to develop versatile bio-photoelectric devices. PMID- 15835977 TI - Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor. AB - Movement of a fluorophore-labeled antibody on the surface of a self-assembled monolayer (SAM) was observed by surface plasmon resonance and surface-plasmon field-enhanced fluorescence spectroscopy (SPFS). At an extremely low surface coverage, the antibody lies close to the biotin-functionalized SAM surface. As additional nonlabeled antibodies were bound, the fluorophore-labeled antibody was displaced away from the SAM surface (and thus the gold surface) due to the constraint imposed by the neighboring nonlabeled antibody. A greater SPFS fluorescence signal was observed due to the weaker fluorescence quenching at large distances from the gold surface. The magnitude of the displacement is proportional to the available biotin binding sites on the sensor surface. The displacement is theoretically explained on the basis of the relationship between the fluorescence intensity and the evanescent field amplitude within the dielectric medium. PMID- 15835978 TI - Correlation between mechanical behavior of protein films at the air/water interface and intrinsic stability of protein molecules. AB - The relation between mechanical film properties of various adsorbed protein layers at the air/water interface and intrinsic stability of the corresponding proteins is discussed. Mechanical film properties were determined by surface deformation in shear and dilation. In shear, fracture stress, sigma(f), and fracture strain, gamma(f), were determined, as well as the relaxation behavior after macroscopic fracture. The dilatational measurements were performed in a Langmuir trough equipped with an infra-red reflection absorption spectroscopy (IRRAS) accessory. During compression and relaxation of the surface, the surface pressure, Pi, and adsorbed amount, Gamma (determined from the IRRAS spectra), were determined simultaneously. In addition, IRRAS spectra revealed information on conformational changes in terms of secondary structure. Possible correlations between macroscopic film properties and intrinsic stability of the proteins were determined and discussed in terms of molecular dimensions of single proteins and interfacial protein films. Molecular properties involved the area per protein molecule at Pi approximately 0 mN/m (A(0)), A(0)/M (M = molecular weight) and the maximum slope of the Pi-Gamma curves (dPi/dGamma). The differences observed in mechanical properties and relaxation behavior indicate that the behavior of a protein film subjected to large deformation may vary widely from predominantly viscous (yielding) to more elastic (fracture). This transition is also observed in gradual changes in A(0)/M. It appeared that in general protein layers with high A(0)/M have a high gamma(f) and behave more fluidlike, whereas solidlike behavior is characterized by low A(0)/M and low gamma(f). Additionally, proteins with a low A(0)/M value have a low adaptability in changing their conformation upon adsorption at the air/water interface. Both results support the conclusion that the hardness (internal cohesion) of protein molecules determines predominantly the mechanical behavior of adsorbed protein layers. PMID- 15835979 TI - Effect of molecular surface packing on the enzymatic activity modulation of an anchored protein on phospholipid Langmuir monolayers. AB - The catalytic activity of a glycosylphosphatidylinositol (GPI)-anchored alkaline phosphatase has been studied in Langmuir phospholipid monolayers at different surface pressures. The enzyme substrate, p-nitrophenyl phosphate, was injected into the subphase of mixed enzyme/lipid Langmuir monolayers. Its hydrolysis product was followed by monitoring the absorbance at 410 nm in situ in the monolayer subphase of the Langmuir trough. Several surface pressures, corresponding to different molecular surface densities, were attained by lateral compression of the monolayers. The morphology of the monolayers, observed by fluorescence microscopy, showed three different types of domains owing to the heterogeneous partition of the enzyme within the mixed enzyme/lipid film. The catalytic activity was modulated by the enzyme surface density, and it increased until a pressure of 18 mN/m was reached, but it decreased significantly when the equilibrium in-plane elasticity (surface compressional modulus) increased more noticeably, resulting in alterations in the interface morphology. A model for the modulation of the enzyme orientation and catalytic activity by lipid/enzyme surface morphology and enzyme surface packing at the air/liquid interface is proposed. The results might have an important impact on the comprehension of the enzymatic activity regulation of GPI-anchored proteins in biomembranes. PMID- 15835980 TI - Force microscopy studies of fibronectin adsorption and subsequent cellular adhesion to substrates with well-defined surface chemistries. AB - Molecular force spectroscopy was used to study the mechanical behavior of plasma fibronectin (FN) on mica, gold, poly(ethylene glycol), and -CH(3), -OH, and -COOH terminated alkanethiol self-assembled monolayers. Proteins were examined at two concentrations, one resulting in a saturated surface with multiple intermolecular interactions referred to as the aggregate state and another resulting in a semiaggregate state where the proteins were neither completely isolated nor completely aggregated. Modeling of the force-extension data using two different theories resulted in similar trends for the fitted thermodynamic parameters from which insight into the protein's binding state could be obtained. Aggregated proteins adsorbed on hydrophobic surfaces adopted more rigid conformations apparently as a result of increased surface denaturation and tighter binding while looser conformations were observed on more hydrophilic surfaces. Studies of FN in a semiaggregate state showed heterogeneity in the model's thermodynamic parameters suggesting that, in the early stages of nonspecific adsorption, multiple protein conformations exist, each having bound irreversibly to the substrate. Proteins in this state all demonstrated a more rigid conformation than in the corresponding aggregate studies due to the greater number of substrate contacts available to the protein. Finally, the force spectroscopy experiments were examined for any biocompatibility correlation by seeding substrates with human umbilical vascular endothelial cells. As predicted from the models used in this work, surfaces with aggregated FN promoted cellular deposition while surfaces with FN in a semiaggregate state appeared to hinder cellular deposition and growth. The atomic force microscope's use as a means for projecting surface biocompatibility, although requiring additional testing, does look promising. PMID- 15835981 TI - Lipid phase transition in saccharide-coated cholate-containing liposomes: coupling to the surrounding matrix. AB - We performed FTIR measurements on cholate-containing liposomes (CCL) embedded in saccharide (trehalose or sucrose) matrixes with different contents of residual water. We obtained information on the CCL phase transition following the thermal evolution (310-70 K) of the IR spectrum of the carbonyl moieties of phospholipids in the frequency range 4225-4550 cm(-1). Furthermore, we simultaneously followed the thermal evolution of the water association band, which gave information on the behavior of the surrounding water-saccharide matrix. The analysis revealed a small sub-band of the water association band present in CCL but not in cholate free liposomes, the thermal evolution of which is tightly coupled to that of the spectrum of the carbonyl moieties of phospholipids. We suggest that this band arises from water molecules, which are inserted within the lipidic structure, in the region located at the border between the hydrophilic and the hydrophobic moieties of phospholipids in the presence of cholic acid. Such water molecules could be responsible for the peculiar flexibility and hydrophilicity of CCL. Following Giuffrida et al. (J. Phys. Chem. B 2003, 107, 13211-13217), we also performed a Spectra Distance analysis, which enabled us to detect an overall liposomes-matrix structural coupling. PMID- 15835982 TI - Nanoscale patterning of alkyl monolayers on silicon using the atomic force microscope. AB - Self-assembled monolayers (SAMs) of 1-alkenes on hydrogen-passivated silicon substrates were successfully patterned on the nanometer scale using an atomic force microscope (AFM) probe tip. Nanoshaving experiments on alkyl monolayers formed on H-Si(111) not only demonstrate the flexibility of this technique but also show that patterning with an AFM probe is a viable method for creating well defined, nanoscale features in a monolayer matrix in a reproducible and controlled manner. Features of varying depths (2-15 nm) were created in the alkyl monolayers by controlling the applied load and the number of etching scans made at high applied loads. The patterning on these SAM films is compared with the patterning of alkyl siloxane monolayers on silicon and mica. PMID- 15835983 TI - Photocatalytic polypyrrole-TiO2-nanoparticles composite thin film generated at the air-water interface. AB - Herein, we report a new method of generation of TiO(2) nanoparticles (NPs) incorporated thin films of polypyrrole (PPy) at the air-water interface. Aqueous TiO(2) NPs when treated with H(2)O(2) and left in a chamber of pyrrole vapor resulted in the formation of a film at the interface, in addition to bulk precipitate. Spectroscopic, X-ray diffraction, and electron microscopic measurements establish the formation of a thin film of PPy with the incorporation of TiO(2) NPs. The TiO(2)-containing PPy films when transferred onto glass substrates were able to photo catalyze the decomposition of aqueous organic dyes: methyl orange and methylene blue. Further, these films could also photo catalyze the oxidation of iodide to triiodide ions in aqueous potassium iodide solution. We find that the PPy-TiO(2) composite films catalyze the reactions in the presence of light more efficiently than a suspension of TiO(2) NPs. PMID- 15835984 TI - Effect of thiocyanate counterion condensation on poly(allylamine hydrochloride) chains on the buildup and permeability of polystyrenesulfonate/polyallylamine polyelectrolyte multilayers. AB - In this study, we investigate the buildup of PEI-(PSS-PAH)(n) polyelectrolyte multilayers at pH 7.4 in the presence of either NaCl or NaSCN as a supporting electrolyte. It appears that in the presence of increasing thiocyanate concentrations (from 0.1 to 0.5 M), the thickness increment, obtained from optical waveguide lightmode spectroscopy experiments, increases whereas it stays practically constant for increasing sodium chloride concentrations (between 0.1 and 0.5 M). The hydration of the films differs also markedly between both electrolyte solutions. The differences in the construction of the polyelectrolyte multilayers in the presence of both supporting electrolytes are rationalized in terms of strong SCN(-) condensation on the PAH chains. The occurrence of this ion condensation is indirectly demonstrated by means of zeta potential measurements and directly demonstrated by means of attenuated total internal reflection infrared spectroscopy on the multilayer films. Moreover when the films are built up in the presence of SCN(-), these ions are only slowly exchanged by the Cl(-) ions introduced in the bulk. Conversely the thick films obtained from 0.5 M NaSCN solutions do not deswell when the buffer solution is replaced by a 0.5 M NaCl containing buffer. The permeability of the films constructed in the presence of both sodium salts is also studied by means of cyclic voltametry and is found to be markedly different in the case of films made from five bilayers at 0.5 M salt concentration. This difference is due to the different morphology and porosity of the films constructed in the presence of 0.5 M NaCl and 0.5 M NaSCN. PMID- 15835985 TI - Proteocubosomes: nanoporous vehicles with tertiary organized fluid interfaces. AB - Proteocubosomes are nanostructured open-nanochannel hierarchical fluid vehicles characterized by a cubic lattice periodicity of the lipid/protein supramolecular assembly (protein-loaded cubosomes). They are obtained here at very high hydration levels by a three-dimensional (3D) self-assembly process, which exploits a protein-directed 3D patterning and fragmentation to create a new, tertiary-level structural order of fluid lipid/water interfaces. Our freeze fracture electron microscopy study reveals that the proteocubosome structures are built up by patterned assemblies of nanocubosomes, which comprise 3D nanoporous fracture surfaces throughout. Complex cubosomic architectures, involving arrays of nanodroplets (larger than 20 nm) inside the proteocubosome particles, are established at high resolution. The soft-matter hierarchical nanocompartment formations display internal aqueous pores belonging to the D-type lipid cubic lattice nanochannel system that is proven by synchrotron X-ray diffraction. The reported nanostructured fluid may give rise to novel applications in nanofluidic biomimetic devices, porous protein drug delivery vehicles, nanoscale enzymatic bioreactors, and protein-encapsulating fluid nanomaterials. PMID- 15835986 TI - Semiflexible polymers confined in soft tubes. AB - We discuss various conformations for a polymer (of persistent length l(p)) confined into a deformable tube (the wall being a lipid bilayer with a certain surface tension sigma and curvature energy K). Our study assumes that there is no adsorption of the chain on the wall. Three states are compared: (a) an unperturbed tube, plus a confined chain, (b) a tube swollen in all the region surrounding the chain (similar to a snake eating a sausage), (c) a globule, a roughly spherical coil surrounded by a strongly deformed tube. We construct a (qualitative) phase diagram for these systems with two variables: the surface tension sigma and the degree of polymerization N. Our main conclusion is that "globules" usually win over "snakes". PMID- 15835987 TI - Self-assembled photoactive polyelectrolyte/perylene-diimide composites. AB - A new class of polyelectrolyte-surfactant (PE-surf) composites having potential applications as thin film organic semiconductors is introduced. These materials are comprised of cationic asymmetrically substituted perylene diimides and oppositely charged poly(acrylate) polyanions. Thin films of the composite materials are prepared by mixing and drop casting aqueous solutions of the two precursors onto appropriate substrates. The resulting materials yield photovoltages of >140 mV for approximately equal to 0.6 W/cm(2) illumination intensities, when incorporated in p-n heterojunction devices. Solution-phase spectra obtained from the PE-surf complexes exhibit excimer-like emission and evidence for formation of weakly coupled aggregates in the ground state. Wide angle X-ray scattering data show the composite films are locally amorphous, while small-angle X-ray data are consistent with a mixture of polymorphic structures that incorporate planar PE-surf bilayers of 3.9-nm repeat distances. Images obtained by conventional far-field light microscopy and multiphoton-excited fluorescence microscopy (MPEFM) indicate that the films are heterogeneous, incorporating submicrometer sized clusters dispersed among much thinner film regions that also incorporate dye. Polarization-dependent MPEFM studies prove the clusters are semiorganized, yielding order parameters (s and P(4)) of 0.09 and 0.01 for in-plane alignment of the chromophores, consistent with a relatively high degree of disorder. PMID- 15835988 TI - Liquid crystalline mesophases of pluronics (L64, P65, and P123) and transition metal nitrate salts ([M(H2O)6](NO3)2). AB - The triblock poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) copolymers, Pluronics (L64, P65, and P123), form liquid crystalline (LC) mesophases with transition metal nitrate salts (TMS), [M(H(2)O)(n)](NO(3))(2), in the presence and absence of free water in the media. In this assembly process, M OH(2) plays an important role as observed in a TMS:C(n)EO(m) (C(n)EO(m) is oligo(ethylene oxide) nonionic surfactants) system. The structure of the LC mesophases and interactions of the metal ion-nitrate ion and metal ion-Pluronic were investigated using microscopy (POM), diffraction (XRD), and spectroscopy (FTIR and micro-Raman) techniques. The TMS:L64 system requires a shear force for mesophase ordering to be observed using X-ray diffraction. However, TMS:P65 and TMS:P123 form well structured LC mesophases. Depending on the salt/Pluronic mole ratio, hexagonal LC mesophases are observed in the TMS:P65 systems and cubic and tetragonal LC mesophases in the TMS:P123 systems. The LC mesophase in the water/salt/Pluronic system is sensitive to the concentration of free (H(2)O) and coordinated water (M-OH(2)) molecules and demonstrates structural changes. As the free water is evaporated from the H(2)O:TMS:Pluronic LC mesophase (ternary mixture), the nitrate ion remains free in the media. However, complete evaporation of the free water molecules enforces the coordination of the nitrate ion to the metal ion in all TMS:Pluronic systems. PMID- 15835989 TI - Pore size engineering in mesoporous silicas using supercritical CO2. AB - In this paper we investigate the use of supercritical carbon dioxide (sc-CO(2)) for synthesizing calcined mesoporous silicas with tunable pore sizes, wall thickness, and d spacings. Small angle neutron scattering was used to probe the controlled swelling of the triblock copolymer surfactant templating agents, P123 (PEO(20)PPO(69)PEO(20)), P85 (PEO(26)PPO(39)PEO(26)), and F127 (PEO(106)PPO(70)PEO(106)), as a function of CO(2) pressure. The transition from the liquid crystal phase to the calcined mesoporous silicas, formed upon condensation and drying, was also studied in detail. Powder X-ray diffraction, transmission electron microscopy, and nitrogen adsorption techniques were used to establish pore diameters, silica wall widths, and the hexagonal packing of the pores within the calcined silicas. Using a direct templating method, the diameters of mesopores and the spacing between the pores could be tuned with a high level of precision. The swelling process was observed to have no detrimental effects on the quality of silica formed, a distinct advantage over conventional swelling techniques, and all of the silicas synthesized in this study were highly ordered over distances of at least 2000 A. PMID- 15835991 TI - Encapsulation of magnetic and fluorescent nanoparticles in emulsion droplets. AB - Oils containing both fluorescent semiconductor and magnetic oxide nanoparticles are used to produce oil in water emulsions. This technique produces oil droplets with homogeneous fluorescence and high magnetic nanoparticle concentrations. The optical properties of the oil droplets are studied as a function of the droplet sizes for various concentrations of fluorescent and magnetic nanoparticles. For all concentrations tested, we find a linear variation of the droplet fluorescent intensity as a function of the droplet volume. For a given size and a given quantum dot (QD) concentration, the droplet fluorescence intensity drops sharply as a function of the magnetic nanoparticle concentration. We show that this decrease is due mainly to the strong absorption cross section of the magnetic nanoparticles and to a lesser extent to the dynamic and static quenching of the QD fluorescence. The role of the iron oxide nanoparticle localization in the droplet (surface versus volume) is also discussed. PMID- 15835990 TI - A novel single-step fabrication technique to create heterogeneous poly(ethylene glycol) hydrogel microstructures containing multiple phenotypes of mammalian cells. AB - In this study, a novel method for the one-step fabrication of stacked hydrogel microstructures using a microfluidic mold is presented. The fabrication of these structures takes advantage of the laminar flow regime in microfluidic devices, limiting the mixing of polymer precursor solutions. To create multilayered hydrogel structures, microfluidic devices were rotated 90 degrees from the traditional xy axes and sealed with a cover slip. Two discreet fluidic regions form in the channels, resulting in the multilayered hydrogel upon UV polymerization. Multilayered patterned poly(ethylene glycol) hydrogel arrays (60 mum tall, 250 mum wide) containing fluorescent dyes, fluorescein isothiocyanate, and tetramethylrhodamine isothiocyanate were created for imaging purposes. Additionally, this method was used to generate hydrogel layers containing murine fibroblasts and macrophages. The cell adhesion promoter, RGD, was added to hydrogel precursor solution to enhance fibroblast cell spreading within the hydrogel matrix in one layer, but not the other. We were able to successfully generate patterns of hydrogels containing multiple phenotypes by using this technique. PMID- 15835992 TI - Assembly of highly aligned DNA strands onto Si chips. AB - This paper reports a robust and efficient approach to assemble highly aligned DNA strands onto Si chips. The method combines advantages from molecular combing and microcontact printing to realize controlling both the density and direction of DNA strands on the Si chip. In addition, it also can be utilized to prepare stretched DNA structures on solid surfaces. Compared to approaches that use molecular combing directly on silanated surfaces, the stretched single-chain DNA structures are straighter. Furthermore, by exploiting the hydrophobic property of the intrinsic poly(dimethylsiloxane) stamp, this study also describes a simple way to produce straight bundled DNA arrays on Si and other substrates. PMID- 15835993 TI - Sonochemical oxidation of multiwalled carbon nanotubes. AB - Functionalization of carbon nanotubes (CNTs) is important for enhancing deposition of metal nanoparticles in the fabrication of supported catalysts. A facile approach for oxidizing CNTs is presented using a sonochemical method to promote the density of surface functional groups. This was successfully employed in a previous study [J. Phys. Chem. B 2004, 108, 19255] to prepare highly dispersed, high-loading Pt nanoparticles on CNTs as fuel cell catalysts. X-ray photoelectron spectroscopy (XPS), transmission electron microscopy, cyclic voltammetry, and settling speeds were used to characterize the degree of surface functionalization and coverage. The sonochemical method effectively functionalized the CNTs. A mixture of -C-O-/-C=O and -COO- was observed along with evidence for weakly bound CO at longer treatment times. The integrated XPS C 1s core level peak area ratios of the oxidized-to-graphitic C oxidation states, as well as the atom % oxygen from the O 1s level, showed an increase in peak intensity (attributed to -CO(x)()) with increased sonication times from 1 to 8 h; the increase in C surface oxidation correlated well with the measured atom %. Most of the CNT surface oxidation occurred between 1 and 2 h. The sonochemically treated CNTs were also studied by cyclic voltammetry and settling experiments, and the results were consistent with the XPS observations. PMID- 15835994 TI - Dissolution and swelling behaviors of random and cross-linked methacrylic acid ethyl acrylate copolymers. AB - The neutralization behaviors of random and cross-linked methacrylic acid-ethyl acrylate (MAA-EA) copolymers were examined as a function of degree of neutralization (alpha) using potentiometric titration and laser light scattering techniques. The random MAA-EA copolymers exhibit a conformational transition from a compact latex particle to a swollen randomly coiled aggregate upon neutralization over a certain range of alpha. With further addition of NaOH, the swollen aggregates dissociate into several smaller clusters. This conformational change is controlled by the balance between electrostatic repulsion within ionized MAA groups and hydrophobic attraction of EA. The cross-linked MAA-EA copolymers do not undergo a drastic conformational change during neutralization. The polymer latex particles swell slightly upon neutralization, and the extent of chain expansion is proportional to MAA molar composition and inversely proportional to cross-linked density. The electrostatic Gibbs energy (DeltaG(el)) obtained from the potentiometric titration data indicates that a higher MAA portion is favorable for the deprotonization of both the random and cross-linked MAA-EA copolymers, suggesting that the dissociation is mainly dominated by polymer structure instead of the electrostatic attraction between H(+) and -COO( ). Moreover, static and dynamic light scattering results confirmed that the cross linked latex particle exists as monodispersed hard sphere in the collapsed state, whereas in its swollen state the latex particle possesses a core-shell structure. PMID- 15835995 TI - Etching of carbon nanotubes by ozone--a surface area study. AB - The oxidative etching of single walled carbon nanotubes (SWNTs) is useful for opening the end caps and the sidewalls of the nanotubes to provide access to the interior. We have studied the effect of successive treatment of SWNTs by 95% pure O(3)(g), which is a powerful and convenient oxidizing agent. The surface area of the SWNTs was measured following exposure to O(3)(g) at 300 K and also following heating to 1073 K in a vacuum to decompose the oxidized groups on the nanotubes, a procedure called etching. This O(3)-induced etching process was observed by scanning electron microscopy and by transmission electron microscopy, and the kinetics of the process was studied gravimetrically. It was found that O(3) attack occurs preferentially on the outermost geometric surface of the conglomerate sample of the nanotubes as a result of the high efficiency of O(3) to react in a few collisions with the nanotube surface. Ozone-induced etching causes the loss of pores in the 20 A diameter range as observed by nitrogen adsorption at equilibrium by density functional theory analysis. PMID- 15835996 TI - Amphiphilic ABC triblock copolymer-assisted synthesis of core/shell structured CdTe nanowires. AB - A new type of amphiphilic ABC triblock copolymer, poly(acrylic acid)(33) poly(styrene)(47)-poly(ethylene oxide)(113) (PAA(33)-PS(47)-PEO(113)), was designed to assist the synthesis of core/shell structured CdTe nanowires via a one-step synthetic route. The PAA block was adopted to capture cadmium ions as the precursor of CdTe. Due to the bivalent coordination of Cd(2+), the copolymer in dioxane/H(2)O formed micelles with Cd(2+)-polychelate cores. Then CdTe nanocrystals were obtained within the micelles after introduction of NaHTe into the micelle solution. Transmission electron microscopy experiments revealed that the CdTe nanocrystals obtained simultaneously formed "pearl-necklace" aggregates in solution possibly driven by dipole interactions between neighboring particles, and then single crystalline CdTe nanowires upon reflux. Accompanying this morphology change, a phase transition from cubic zinc blende to wurtzite structure was observed by selected-area electron diffraction. The aggregation of the PS block in dioxane with a certain amount of H(2)O enabled the PS blocks to form a densely packed shell on the CdTe nanowires whose typical size is 700-800 nm in length and 15-20 nm in width. The third block of PEO was employed to render the finally formed CdTe nanowires dispersibility. PMID- 15835997 TI - Electrowetting dynamics of microfluidic actuation. AB - When voltage is suddenly applied to vertical, parallel dielectric-coated electrodes dipped into a liquid with finite conductivity, the liquid responds by rising up to reach a new hydrostatic equilibrium height. On the microfluidic scale, the dominating mechanism impeding this electromechanically induced actuation appears to be a dynamic friction force that is directly proportional to the velocity of the contact line moving along the solid surface. This mechanism has its origin in the molecular dynamics of the liquid coming into contact with the solid surface. A simple reduced-order model for the rising column of liquid is used to quantify the magnitude of this frictional effect by providing estimates for the contact line friction coefficient. Above some critical threshold of voltage, the electromechanical force is clamped, presumably by the same mechanism responsible for contact angle saturation and previously reported static height-of-rise limits. The important distinction for the dynamic case is that the onset of the saturation effect is delayed in time until the column has risen more than about halfway to its static equilibrium height. PMID- 15835998 TI - Molecular simulation study of the effect of pressure on the vapor-liquid interface of the square-well fluid. AB - We examine a model system to study the effect of pressure on the surface tension of a vapor-liquid interface. The system is a two-component mixture of spheres interacting with the square-well (A-A) and hard-sphere (B-B) potentials and with unlike (A-B) interactions ranging (for different cases) from hard sphere to strongly attractive square well. The bulk-phase and interfacial properties are measured by molecular dynamics simulation for coexisting vapor-liquid phases for various mixture compositions, pressures, and temperatures. The variation of the surface tension with pressure compares well to values given by surface-excess formulas derived from thermodynamic considerations. We find that surface tension increases with pressure only for the case of an inert solute (hard-sphere A-B interactions) and that the presence of A-B attractions strongly promotes a decrease of surface tension with pressure. An examination of density and composition profiles is made to explain these effects in terms of surface adsorption arguments. PMID- 15835999 TI - Simulating the equation of state of model globular proteins adsorbed at a surface. AB - Monte Carlo computer simulation is used to follow the adsorption of a model globular and disordered protein at a hard surface and to simulate the surface equation of state for these molecules. The simulation utilizes the deformable globule model where the "protein" is treated as a collection of interacting subunits. Disordered globules are modeled as athermal molecules, whereas globular molecules have a strong attractive interaction between subunits. The surface equation of state is modeled by applying a known pressure to an adsorbed globule and following the changes in adsorbed conformation. Simulated equations of state for the disordered and globular molecules show features that are observed in experimental surface pressure versus area plots. In particular the simulated equations of state show "kinks" that correspond to regions where the adsorbed globules undergo conformational changes as they lift away from the surface in response to the increased pressure. The model proteins follow Bull's equation at low surface pressures in a way that is broadly in line with results from experiment, and the changes in conformation as a function of surface pressure are in line with predictions by DeFeijter and Benjamins made using a soft-particle model for adsorbed proteins. PMID- 15836000 TI - Measuring the surface roughness of sputtered coatings by microgravimetry. AB - A novel application of quartz crystal microgravimetry is described to enable determination of the roughness of native and sputtered oxide coatings. The technique is applicable to coatings that may be too rough for accurate atomic force microscopic imaging where measurements may be limited by the dimensions of the probe tip. PMID- 15836001 TI - Vibration-actuated drop motion on surfaces for batch microfluidic processes. AB - When a liquid drop is subjected to an asymmetric lateral vibration on a nonwettable surface, a net inertial force acting on the drop causes it to move. The direction and velocity of the drop motion are related to the shape, frequency, and amplitude of vibration, as well as the natural harmonics of the drop oscillation. Aqueous drops can be propelled through fluidic networks connecting various unit operations in order to carry out batch processing at the miniature scale. We illustrate the integration of several unit operations on a chip: drop transport, mixing, and thermal cycling, which are precursor steps to carrying out advanced biological processes at microscale, including cell sorting, polymerase chain reaction, and DNA hybridization. PMID- 15836002 TI - EMEA: for patients or for industry? PMID- 15836003 TI - The relationship between productivity and health-related QOL: an exploration. AB - In economic evaluation of healthcare programmes both QOL and productivity of patients are aspects to be studied. Normally, the former is part of the measurement of the effectiveness of the programme and the latter is part of the measurement of its costs. In this paper we highlight the relationship between QOL and productivity. Two aspects are discussed: (i) the relationship between perceived productivity and health-state valuations; and (ii) the observed relationship between productivity and QOL. The first aspect relates to the fact that in health-state valuations, respondents may consider income changes and ability to work. While this may have important methodological and practical implications, little empirical evidence exists in this area. The second aspect relates to the fact that the observed productivity of individuals is expected to be related to their health-related QOL. Worse health states are expected to be associated with lower productivity. Again, empirical investigation is lacking; however, this relationship may prove important, for instance in modelling productivity costs with use of information on QOL. This paper explores these relationships between productivity and QOL to stimulate debate and research in this area. PMID- 15836004 TI - Penny and pound wise: pharmacoeconomics from a governmental perspective. AB - Because of growing pressure on the healthcare budget in The Netherlands, appropriate justification of current expenditures and future investments in public healthcare are becoming increasingly important. Therefore, the Dutch Ministry of Health, Welfare and Sport is expanding its use of pharmacoeconomic evaluation in informed reimbursement decision-making of new pharmaceuticals. Since June 2002, pharmaceutical companies have been invited to submit a pharmacoeconomic dossier with their reimbursement applications of innovative drugs. As of January 2005 submission of a pharmacoeconomic dossier is mandatory for all drugs claiming to have therapeutic value. Currently, several European governmental and non-governmental organisations are making efforts to harmonise pharmacoeconomic research guidelines at the EU level. Ultimately, this may facilitate a more efficient way of conducting pharmacoeconomic research and encourage the use of pharmacoeconomic data by national assessment agencies and governments. It is anticipated that international pharmaceutical companies will increasingly invest in pharmacoeconomics while government staff will become more experienced in appraising the dossiers, thus resulting in an upward momentum in the quality and usability of pharmacoeconomic data. From the Dutch government's perspective, the use of pharmacoeconomic evaluation in reimbursement decision making should offer a true opportunity for pharmaceutical companies to present the added value for money of new drugs. Using pharmacoeconomic data, costs, benefits and effects of pharmaceuticals are increasingly being considered from a societal perspective, thus going beyond the sole consideration of the impact on the pharmaceutical budget. PMID- 15836005 TI - The economic impact of neuropsychiatric symptoms in Alzheimer's disease: can drugs ease the burden? AB - The majority of patients with Alzheimer's disease (AD) will have clinically significant neuropsychiatric symptoms during the course of their disease. There is growing evidence that neuropsychiatric symptoms increase direct costs of care in patients with AD, especially the costs associated with formal long-term care and unpaid caregiving. For example, we have estimated that a 1-point worsening of the neuropsychiatric inventory score is associated with an incremental increase of between USD 247 and USD 409 per year in total direct costs of care based upon year 2001 US dollars, depending on the value of unpaid caregiving. Although data are still limited, there have been a series of well designed, controlled clinical trials that have established the efficacy of several drugs used in the treatment of neuropsychiatric symptoms in patients with AD. The economic impact of using efficacious drugs to treat neuropsychiatric symptoms in patients with AD has not been evaluated formally. To successfully complete formal economic evaluations of these drugs there is a need for more research to refine methods for determining the economic value of unpaid caregiving and to collect more data concerning the incremental effects of neuropsychiatric symptoms on QOL, costs of care and survival. The current ongoing treatment trials that are collecting economic and QOL data as a part of the trial will be able to perform cost-effectiveness and cost-utility analyses of these new efficacious drugs. These economic evaluations will provide important information for decision makers who are formulating healthcare policy for the treatment of patients with AD. PMID- 15836006 TI - International variation in resource utilisation and treatment costs for rheumatoid arthritis: a systematic literature review. AB - Recent years have witnessed substantial progress in understanding the cost implications of rheumatoid arthritis (RA). To assess the divergent methodologies and their impact on the resulting cost analyses in RA, we conducted a systematic literature review to summarise the scientific evidence of RA-induced costs. Sixty five reviews, models or cost analyses on the burden of illness and general costs associated with RA were identified. They covered the US, Canada, Sweden, the UK, The Netherlands, Germany and Finland. Twenty-four cost analyses provided appropriate data about direct and/or indirect costs. Each study was summarised separately. Costs were discounted to 2003 and converted to US dollars. The costs per RA-year ranged from USD 1503 to USD 16,514. However, each study has to be interpreted individually, with consideration given to the study population, indication, age of the study, database used, type of therapy, setting, level of cost differentiation and data derivation. Health technology assessment reports offer sufficient space to adequately describe the composite parts and restrictive elements of different methodological approaches and analyses. PMID- 15836007 TI - Oral serotonin receptor agonists: a review of their cost effectiveness in migraine. AB - Migraine headache is a highly prevalent chronic, episodic condition. The direct and indirect costs of migraine headache have a tremendous economic impact in the US. Research has shown that serotonin (5HT(1B/D)) receptor agonists reduce healthcare costs, improve health-related QOL (HR-QOL), decrease migraine disability and keep patients effective in the workplace. The purpose of this manuscript is to examine the cost effectiveness of oral 5HT(1B/D) receptor agonists for the treatment of migraine headache. In general, 5HT(1B/D) receptor agonists are associated with increases in direct healthcare costs; however, they are also associated with reductions in the indirect costs associated with migraine headache. Therefore, it appears that the relatively high acquisition cost of these medications is offset and, as a class, these medications appear to be cost effective and demonstrate net benefits from the societal perspective. Based on meta-analyses in which data on eletriptan were not available, it appears that within the class, almotriptan and rizatriptan are the most cost effective. In a prospective study comparing eletriptan with sumatriptan, it appears that the former may be more cost effective than the latter. Additional investigations are needed to further explore the application of the friction-cost approach and QALYs to cost-effectiveness analyses of this class of medication. PMID- 15836008 TI - A pharmacoeconomic evaluation of statins in the treatment of hypercholesterolaemia in the primary care setting in Spain. AB - BACKGROUND: Cardiovascular disease is one of the leading causes of death and it has been shown that primary prevention with the HMG-CoA reductase inhibitor (statin) lipid-lowering drugs can reduce cardiovascular events. Acquisition costs vary between statins and this may be an important consideration in the overall cost effectiveness (CE) of different options. OBJECTIVE: To perform a CE study of the main statins used in Spain for primary prevention of cardiovascular disease in patients with high cholesterol levels [corrected] STUDY DESIGN: The CE analysis was based on an open-label, prospective, naturalistic, randomised intervention study under usual care conditions in primary care settings in patients with high cholesterol levels (total cholesterol [TC] >240 mg/dL, low density lipoprotein cholesterol [LDL-C] >160 mg/dL) and one or more cardiovascular risk factors. The analysis was conducted from the perspective of the Spanish National Health System; the year of costing was 2001. PATIENTS: A total of 161 patients (49.7% males), mean age 65 +/- 10.3 years, without evidence of cardiovascular disease were included in the study. Of those, 82.1% were hypertensive, 37.1% had diabetes mellitus and 17.9% were smokers. INTERVENTIONS: Forty-eight patients received oral atorvastatin 10 mg/day, 32 received fluvastatin 40 mg/day, 44 received simvastatin 20 mg/day and 37 patients received pravastatin 20 mg/day for 6 months. MAIN MEASUREMENTS AND RESULTS: After 6 months, the therapeutic goals of LDL-C control, according to the recommendations of the Spanish Society of Arteriosclerosis--Consensus-2000, were reached in 62.5%, 43.8%, 45.5% and 40.5% of patients treated with atorvastatin, fluvastatin, simvastatin and pravastatin, respectively. The average CE ratio, expressed as the cost in euros (euro) per patient achieving the therapeutic goals, was euros 424.3 for atorvastatin, euros 503.5 for fluvastatin, euros 527.0 for simvastatin and euros 683.4 for pravastatin. The incremental CE ratios for atorvastatin versus fluvastatin and simvastatin were euros 238.9 and euros 149.5, respectively, per additional patient reaching therapeutic goals. Atorvastatin, fluvastatin and simvastatin all dominated pravastatin. CONCLUSIONS: All the statins studied have been shown to be effective for reducing both TC and LDL-C levels. In this study, atorvastatin was the most efficient drug, with the best CE ratio (cost per patient reaching therapeutic goals). Atorvastatin was more effective and less costly than pravastatin, and when compared with fluvastatin or simvastatin the additional cost per additional patient achieving therapeutic goals was 2 actually decrease upon strengthening of correlation. The explicit formulas for l-matrices and {n(l)} can be readily utilized in testing, calibration, and benchmarking of approximate implementations of the density matrix functional theory (DMFT). PMID- 15836016 TI - Corrections to the law of mass action and properties of the asymptotic t=infinity state for reversible diffusion-limited reactions. AB - For diffusion-limited reversible A+A<==>B reactions we reexamine two fundamental concepts of classical chemical kinetics-the notion of "chemical equilibrium" and the "law of mass action." We consider a general model with distance-dependent reaction rates, such that any pair of A particles, performing standard random walks on sites of a d-dimensional lattice and being at a distance mu apart of each other at time moment t, may associate forming a B particle at the rate k+(mu). In turn, any randomly moving B particle may spontaneously dissociate at the rate k-(lambda) into a geminate pair of As "born" at a distance lambda apart of each other. Within a formally exact approach based on Gardiner's Poisson representation method we show that the asymptotic t=infinity state attained by such diffusion-limited reactions is generally not a true thermodynamic equilibrium, but rather a nonequilibrium steady state, and that the law of mass action is invalid. The classical concepts hold only in case when the ratio k+(mu)k-(mu) does not depend on mu for any mu. PMID- 15836017 TI - Ab initio potential-energy surfaces for complex, multichannel systems using modified novelty sampling and feedforward neural networks. AB - A neural network/trajectory approach is presented for the development of accurate potential-energy hypersurfaces that can be utilized to conduct ab initio molecular dynamics (AIMD) and Monte Carlo studies of gas-phase chemical reactions, nanometric cutting, and nanotribology, and of a variety of mechanical properties of importance in potential microelectromechanical systems applications. The method is sufficiently robust that it can be applied to a wide range of polyatomic systems. The overall method integrates ab initio electronic structure calculations with importance sampling techniques that permit the critical regions of configuration space to be determined. The computed ab initio energies and gradients are then accurately interpolated using neural networks (NN) rather than arbitrary parametrized analytical functional forms, moving interpolation or least-squares methods. The sampling method involves a tight integration of molecular dynamics calculations with neural networks that employ early stopping and regularization procedures to improve network performance and test for convergence. The procedure can be initiated using an empirical potential surface or direct dynamics. The accuracy and interpolation power of the method has been tested for two cases, the global potential surface for vinyl bromide undergoing unimolecular decomposition via four different reaction channels and nanometric cutting of silicon. The results show that the sampling methods permit the important regions of configuration space to be easily and rapidly identified, that convergence of the NN fit to the ab initio electronic structure database can be easily monitored, and that the interpolation accuracy of the NN fits is excellent, even for systems involving five atoms or more. The method permits a substantial computational speed and accuracy advantage over existing methods, is robust, and relatively easy to implement. PMID- 15836018 TI - Characterizing the network topology of the energy landscapes of atomic clusters. AB - By dividing potential energy landscapes into basins of attractions surrounding minima and linking those basins that are connected by transition state valleys, a network description of energy landscapes naturally arises. These networks are characterized in detail for a series of small Lennard-Jones clusters and show behavior characteristic of small-world and scale-free networks. However, unlike many such networks, this topology cannot reflect the rules governing the dynamics of network growth, because they are static spatial networks. Instead, the heterogeneity in the networks stems from differences in the potential energy of the minima, and hence the hyperareas of their associated basins of attraction. The low-energy minima with large basins of attraction act as hubs in the network. Comparisons to randomized networks with the same degree distribution reveals structuring in the networks that reflects their spatial embedding. PMID- 15836019 TI - Chemical reaction rates from ring polymer molecular dynamics. AB - We show how the ring-polymer molecular dynamics method can be adapted to calculate approximate Kubo-transformed flux-side correlation functions, and hence rate coefficients for condensed phase reactions. An application of the method to the standard model for a chemical reaction in solution--a quartic double-well potential linearly coupled to a bath of harmonic oscillators--is found to give results of comparable accuracy to those of the classical Wigner model and the centroid molecular dynamics method. However, since the present method does not require that one evaluate the Wigner transform of a thermal flux operator or that one perform a separate path integral calculation for each molecular dynamics time step, we believe it will prove easier to apply to more general problems than either of these alternative techniques. We also present a (logarithmic) discretization scheme for the Ohmic bath in the system-bath model that gives converged results with just nine bath modes--a surprisingly small number for a model of a condensed phase reaction. Finally, we present some calculations of the transmission through an Eckart barrier which show that the present method provides a satisfactory (although not perfect) description of the deep quantum tunneling regime. Part of the reason for the success of the method is that it gives the exact quantum-mechanical rate constant for the transmission through a parabolic barrier, as we demonstrate analytically in the Appendix. PMID- 15836020 TI - Coupled-cluster theory with simplified linear-r(12) corrections: the CCSD(R12) model. AB - A simplified singles-and-doubles linear-r(12) corrected coupled-cluster model, denoted CCSD(R12), is proposed and compared with the complete singles-and-doubles linear-r(12) coupled-cluster method CCSD-R12. An orthonormal auxiliary basis set is used for the resolution-of-the-identity approximation to calculate three electron integrals needed in the linear-r(12) Ansatz. Basis-set convergence is investigated for a selected set of atoms and small molecules. In a large basis, the CCSD(R12) model provides an excellent approximation to the full linear-r(12) energy contribution, whereas the magnitude of this contribution is significantly overestimated at the level of second-order perturbation theory. PMID- 15836021 TI - Scalar relativistic all-electron density functional calculations on periodic systems. AB - Scalar relativistic effects are included in periodic boundary conditions calculations with Gaussian orbitals. This approach is based on the third-order Douglas-Kroll-Hess approximation, allowing the treatment of all electrons on an equal footing. With this methodology, we are able to perform relativistic all electron density functional calculations using the traditional local spin-density and generalized gradient approximations (GGA), as well as meta-GGA and hybrid density functionals. We present benchmark results for the bulk metals Pd, Ag, Pt, and Au, and the large band gap semiconductors AgF and AgCl. PMID- 15836022 TI - Alchemical free energy calculations and multiple conformational substates. AB - Thermodynamic integration (TI) was combined with (adaptive) umbrella sampling to improve the convergence of alchemical free energy simulations in which multiple conformational substates are present. The approach, which we refer to as non Boltzmann TI (NBTI), was tested by computing the free energy differences between three five-atomic model systems, as well as the free energy difference of solvation between leucine and asparagine. In both cases regular TI failed to give converged results, whereas the NBTI results were free from hysteresis and had standard deviations well below +/-0.7 kcal/mole. We also present theoretical considerations that make it possible to compute free energy differences between simple molecules, such as the five-atomic model systems, by numerical integration of the partition functions at the respective end points. PMID- 15836023 TI - Quantum control of molecular motion including electronic polarization effects with a two-stage toolkit. AB - A method for incorporating strong electric field polarization effects into optimal control calculations is presented. A Born-Oppenheimer-type separation, referred to as the electric-nuclear Born-Oppenheimer (ENBO) approximation, is introduced in which variations of both the nuclear geometry and the external electric field are assumed to be slow compared with the speed at which the electronic degrees of freedom respond to these changes. This assumption permits the generation of a potential energy surface that depends not only on the relative geometry of the nuclei but also on the electric field strength and on the orientation of the molecule with respect to the electric field. The range of validity of the ENBO approximation is discussed in the paper. A two-stage toolkit implementation is presented to incorporate the polarization effects and reduce the cost of the optimal control dynamics calculations. As an illustration of the method, it is applied to optimal control of vibrational excitation in a hydrogen molecule aligned along the field direction. Ab initio configuration interaction calculations with a large orbital basis set are used to compute the H-H interaction potential in the presence of the electric field. The significant computational cost reduction afforded by the toolkit implementation is demonstrated. PMID- 15836024 TI - Is the Filinov integral conditioning technique useful in semiclassical initial value representation methods? AB - The utility of the Filinov integral conditioning technique, as implemented in semiclassical initial value representation (SC-IVR) methods, is analyzed for a number of regular and chaotic systems. For nonchaotic systems of low dimensionality, the Filinov technique is found to be quite ineffective at accelerating convergence of semiclassical calculations since, contrary to the conventional wisdom, the semiclassical integrands usually do not exhibit significant phase oscillations in regions of large integrand amplitude. In the case of chaotic dynamics, it is found that the regular component is accurately represented by the SC-IVR, even when using the Filinov integral conditioning technique, but that quantum manifestations of chaotic behavior was easily overdamped by the filtering technique. Finally, it is shown that the level of approximation introduced by the Filinov filter is, in general, comparable to the simpler ad hoc truncation procedure introduced by Kay. PMID- 15836025 TI - Laser control of electronic transitions of wave packet by using quadratically chirped pulses. AB - An effective scheme is proposed for the laser control of wave packet dynamics. It is demonstrated that by using specially designed quadratically chirped pulses, fast and nearly complete excitation of wave packet can be achieved without significant distortion of its shape. The parameters of the laser pulse can be estimated analytically from the Zhu-Nakamura theory of nonadiabatic transition. If the wave packet is not too narrow or not too broad, then the scheme is expected to be utilizable for multidimensional systems. The scheme is applicable to various processes such as simple electronic excitation, pump-dump, and selective bond breaking, and it is actually numerically demonstrated to work well by taking diatomic and triatomic molecules (LiH, NaK, H(2)O) as examples. PMID- 15836026 TI - Quantum fluctuation of electronic wave-packet dynamics coupled with classical nuclear motions. AB - An ab initio electronic wave-packet dynamics coupled with the simultaneous classical dynamics of nuclear motions in a molecule is studied. We first survey the dynamical equations of motion for the individual components. Reflecting the nonadiabatic dynamics that electrons can respond to nuclear motions only with a finite speed, the equations of motion for nuclei include a force arising from the kinematic (nuclear momentum) coupling from electron cloud. To materialize these quantum effects in the actual ab initio calculations, we study practical implementation of relevant electronic matrix elements that are related to the derivatives with respect to the nuclear coordinates. Applications of the present scheme are performed in terms of the configuration state functions (CSF) using the canonical molecular orbitals as basis functions without transformation to particular diabatic basis. In the CSF representation, the nonadiabatic interaction due to the kinematic coupling is anticipated to be rather small, and instead it should be well taken into account through the off-diagonal elements of the electronic Hamiltonian matrix. Therefore it is expected that the nonadiabatic dynamics based on this CSF basis neglecting the kinematic coupling may work. To verify this anticipation and to quantify the actual effects of the kinematic coupling, we compare the dynamics with and without the kinematic-coupling terms using the same CSF set. Applications up to the fifth electronically excited states in a nonadiabatic collision between H(2) and B(+) shows that the overall behaviors of these two calculations are surprisingly similar to each other in an average sense except for a fast fluctuation reflecting the electronic time scale. However, at the same time, qualitative differences in the collision events are sometimes observed. Therefore it turns out after all that the kinematic-coupling terms cannot be neglected in the CSF-basis representation. The present applications also demonstrate that the nonadiabatic electronic wave-packet dynamics within ab initio quantum chemical calculation is feasible. PMID- 15836027 TI - Comparison of two genres for linear scaling in density functional theory: purification and density matrix minimization methods. AB - Two classes of linear-scaling methods to replace diagonalization of the one particle Hamiltonian matrix in density functional theory are compared to each other. Purification takes a density matrix with the correct eigenfunctions and corrects the occupation numbers; density matrix minimization takes a density matrix with correct occupation numbers and corrects the eigenfunctions by rotating the orbitals. Computational comparisons are performed through modification of the MondoSCF program on water clusters and the protein endothelin. A purification scheme and a density matrix minimization scheme, based on the 1,2-contracted Schrodinger equation [D. A. Mazziotti, J. Chem. Phys. 115, 8305 (2001)] are implemented in large systems. PMID- 15836028 TI - Correlation and response functions with non-Markovian dissipation: a reduced Liouville-space theory. AB - Based on a recently developed quantum dissipation formulation [R. X. Xu and Y. J. Yan, J. Chem. Phys. 116, 9196 (2002)], we present a reduced Liouville-space approach to evaluate the response and correlation functions of dissipative systems. The weak system-bath interaction is treated properly for its effects on the initial state, the evolution, and the correlation between coherent driving and non-Markovian dissipation. Numerical demonstration shows this correlated effect cannot be neglected even in the calculation of linear response quantities that do not explicitly depend on external fields. Highlighted in this paper is also the proper choice of theory among various formulations in the weak system bath interaction regime. PMID- 15836029 TI - Orbital-optimized coupled-cluster theory does not reproduce the full configuration-interaction limit. AB - It is shown that due to the mixing of the usual projection approach of coupled cluster with variational orbital optimization, orbital-optimized coupled cluster (OCC) fails to reproduce the full configuration-interaction (full CI) limit when the cluster operator becomes complete. It is pointed out that the fulfillment of the projected singles equations, which define the orbital gradient in Brueckner coupled cluster (BCC), is mandatory for a correct behavior. As numerical examples we present general OCC and BCC calculations up to the full CI limit on CH(2) and an active-space model of ozone. The observed deviations of OCC from full CI are of the order of the correlation error obtained in calculations with up to quadruples excitations. Thus the failure of OCC may be considered tolerable in more approximate calculations but clearly prohibitive for any benchmark application. For applications to active-space models a hybrid approach for OCC is suggested in which for active particle-hole rotations the Brueckner orbital gradient is employed, whereas for the remaining orbital rotations the variational orbital gradient is retained. PMID- 15836030 TI - Correlated holes and their relationships with reduced density matrices and cumulants. AB - This paper describes a matrix formulation for the correlated hole theory within the framework of the domain-averaged model in many electron systems (atoms, molecules, condensed matter, etc.). General relationships between this quantity and one-particle reduced density matrices for any independent particle or correlated state functions are presented. This formulation turns out to be suitable for computational purposes due to the straightforward introduction of cumulants of two-particle reduced density matrices within the quantum field structure. Numerical calculations in selected simple molecular systems have been performed in order to determine preliminary correlated values for such a quantity. PMID- 15836031 TI - Stabilization and relative phase effects in a dichromatically driven diatomic Morse molecule: interpretation based on nonlinear classical dynamics. AB - The dissociation dynamics of a dichromatically laser-driven diatomic Morse molecule vibrating in the ground state is investigated by applying tools of the nonlinear theory of classical Hamiltonian systems. Emphasis is placed on the role of the relative phase of the two fields, phi. First, it is found that, just like in quantum mechanics, there is dependence of the dissociation probability on phi. Then, it is demonstrated that addition of the second laser leads to suppression of probability (stabilization), when the intensity of the first laser is kept constant just above or below the single laser dissociation threshold. This "chemical bond hardening" diminishes as phi increases. These effects are investigated and interpreted in terms of modifications in phase space topology. Variations of phi as well as of the intensity of the second laser may cause (i) appearance/disappearance of the stability island corresponding to the common resonance with the lowest energy and (ii) deformation and movement of the region of Kolmogorov-Arnold-Moser tori that survive from the undriven system. The latter is the main origin in phase space of stabilization and phi dependence. Finally, it is shown that the use of short laser pulses enhances both effects. PMID- 15836032 TI - Introducing ONETEP: linear-scaling density functional simulations on parallel computers. AB - We present ONETEP (order-N electronic total energy package), a density functional program for parallel computers whose computational cost scales linearly with the number of atoms and the number of processors. ONETEP is based on our reformulation of the plane wave pseudopotential method which exploits the electronic localization that is inherent in systems with a nonvanishing band gap. We summarize the theoretical developments that enable the direct optimization of strictly localized quantities expressed in terms of a delocalized plane wave basis. These same localized quantities lead us to a physical way of dividing the computational effort among many processors to allow calculations to be performed efficiently on parallel supercomputers. We show with examples that ONETEP achieves excellent speedups with increasing numbers of processors and confirm that the time taken by ONETEP as a function of increasing number of atoms for a given number of processors is indeed linear. What distinguishes our approach is that the localization is achieved in a controlled and mathematically consistent manner so that ONETEP obtains the same accuracy as conventional cubic-scaling plane wave approaches and offers fast and stable convergence. We expect that calculations with ONETEP have the potential to provide quantitative theoretical predictions for problems involving thousands of atoms such as those often encountered in nanoscience and biophysics. PMID- 15836033 TI - Photodissociation of HI and DI: polarization of atomic photofragments. AB - The complete angular momentum distributions and vector correlation coefficients (orientation and alignment) of ground state I((2)P(32)) and excited state I((2)P(12)) atoms resulting from the photodissociation of HI have been computed as a function of photolysis energy. The orientation and alignment parameters a(Q) ((K))(p) that describe the coherent and incoherent contributions to the angular momentum distributions from the multiple electronic states accessed by parallel and perpendicular transitions are determined using a time-dependent wave packet treatment of the dissociation dynamics. The dynamics are based on potential energy curves and transition dipole moments that have been reported previously [R. J. LeRoy, G. T. Kraemer, and S. Manzhos, J. Chem. Phys. 117, 9353 (2002)] and used to successfully model the scalar (total cross section and branching fraction) and lowest order vector (anisotropy parameter beta) properties of the photodissociation. Predictions of the a(Q) ((K))(p), parameters for the isotopically substituted species DI are reported and contrasted to the analogous HI results. The resulting polarization for the corresponding H/D partners are also determined and demonstrate that both H and D atoms produced can be highly spin polarized. Comparison of these predictions for HI and DI with experimental measurement will provide the most stringent test of the current model for the electronic structure and the interpretation of the dissociation based on noncoupled excited state dynamics. PMID- 15836034 TI - Iron monoxide photodissociation. AB - The photodissociation of (56)FeO was studied by means of the velocity map imaging technique. A molecular beam of iron atoms and iron monoxide molecules was created using an electrical discharge with an iron electrode in a supersonic expansion of molecular oxygen. The ground state iron atom Fe((5)D(4)) and FeO concentrations in the molecular beam have been estimated. The dissociation energy of the FeO X (5)Delta ground electronic state was found to be D(0) (0)(FeO)=4.18+/-0.01 eV. The effective absorption cross section of FeO at 252.39 nm (vac), leading to the Fe((5)D(4))+O((3)P) dissociation channel, is approximately 1.2 x 10(-18) cm(2). A (1+1) resonantly enhanced multiphoton ionization spectrum of (56)FeO in the region 39 550-39 580 cm(-1) with rotational structure has been observed, but not assigned. Angular distributions of Fe((5)D(4)) and Fe((5)D(3)) products for the channel FeO-->Fe((5)D(4,3))+O((3)P) have been measured at several points in the 210-260 nm laser light wavelength region. The anisotropy parameter varies strongly with wavelength for both channels. PMID- 15836035 TI - Effect of bending and torsional mode excitation on the reaction Cl+CH4-->HCl+CH3. AB - A beam containing CH(4), Cl(2), and He is expanded into a vacuum chamber where CH(4) is prepared via infrared excitation in a combination band consisting of one quantum of excitation each in the bending and torsional modes (nu(2)+nu(4)). The reaction is initiated by fast Cl atoms generated by photolysis of Cl(2) at 355 nm, and the resulting CH(3) and HCl products are detected in a state-specific manner using resonance-enhanced multiphoton ionization (REMPI). By comparing the relative amplitudes of the action spectra of Cl+CH(4)(nu(2)+nu(4)) and Cl+CH(4)(nu(3)) reactions, we determine that the nu(2)+nu(4) mode-driven reaction is at least 15% as reactive as the nu(3) (antisymmetric stretch) mode-driven reaction. The REMPI spectrum of the CH(3) products shows no propensity toward the formation of umbrella bend mode excited methyl radical, CH(3)(nu(2)=1), which is in sharp distinction to the theoretical expectation based on adiabatic correlations between CH(4) and CH(3). The rotational distribution of HCl(v=1) products from the Cl+CH(4)(nu(2)+nu(4)) reaction is hotter than the corresponding distribution from the Cl+CH(4)(nu(3)) reaction, even though the total energies of the two reactions are the same within 4%. An explanation for this enhanced rotational excitation of the HCl product from the Cl+CH(4)(nu(2)+nu(4)) reaction is offered in terms of the projection of the bending motion of the CH(4) reagent onto the rotational motion of the HCl product. The angular distributions of the HCl(nu=0) products from the Cl+CH(4)(nu(2)+nu(4)) reaction are backward scattered, which is in qualitative agreement with theoretical calculation. Overall, nonadiabatic product vibrational correlation and mode specificity of the reaction indicate that either the bending mode or the torsional mode or both modes are strongly coupled to the reaction coordinate. PMID- 15836036 TI - Bonding behavior and thermal stability of C54Si6: a first-principles molecular dynamics study. AB - By using a self-consistent plane-waves density functional approach we study the bonding behavior and the thermal properties of C(54)Si(6) heterofullerenes. Calculations are carried out by employing a generalized gradient approximation. Our investigation improves upon previous findings on the same system obtained via the local density approximation approach. This is due to a much larger search in configurational space and the explicit account of temperature effects. Overall, isomers can be classified in two groups. In the first, nearest-neighboring Si atoms form a subnetwork, while in the second the Si atoms are farther apart on the cage. In addition to structural optimization, we carried out first-principles molecular dynamics for temperatures up to T=3000 K on a time interval of 12 ps. These simulations show that Si-Si bond variations with temperature are less important when all Si atoms are found on a hexagon. Therefore, this structural arrangement is the most likely to be observed experimentally. Analysis of charge topology reveals that the amount of charge on each atom depends on the number of heterogeneous bonds, due to a significant charge transfer from the Si to the neighboring C atoms. PMID- 15836037 TI - Photodissociation spectroscopy of Zn+(H2O) and Zn+(D2O). AB - We report on a study of the photodissociation spectroscopy of weakly bound Zn+(H2O) and Zn+(D2O) complexes. The work is supported by ab initio electronic structure calculations of the ground and low-lying excited energy surfaces. We assign two molecular absorption bands in the near UV correlating to Zn+ (4s-4p) based transitions, and identify vibrational progressions associated with both intermolecular and intramolecular vibrational modes of the cluster. Partially resolved rotational structure is consistent with a C(2V) equilibrium complex geometry. Experimental spectroscopic constants are in very good agreement with ab initio theoretical predictions. Results are compared with previous work on main group and transition metal ion-H2O clusters. PMID- 15836038 TI - Angular distribution of different vibrational components of the X and B states reached after resonant Auger decay of core-excited H2O: experiment and theory. AB - Vibrationally resolved spectra have been obtained for the lowest-lying cationic states X (2)B(1), A (2)A(1), and B (2)B(2) of the water molecule reached after participator resonant Auger decay of core-excited states. The angular distribution has been measured of the first four vibrational components of the X state in the photon energy regions including the O 1s-->4a(1) and the O 1s- >2b(2) core excitations, and for different portions of the vibrational envelope of the B state in the photon energy region including the O 1s-->2b(2) core excitation. For the X state, a large relative spread in beta values of the different vibrational components is observed across both resonances. For the B state, a very different trend is observed for the high binding energy side and the low binding energy side of the related spectral feature as a function of photon energy. A theoretical method based on the scattering K matrix has been used to calculate both the photoabsorption spectrum and the beta values, by taking both interference between direct and resonant photoemission and vibrational/lifetime interference into account. The numerical results show qualitative agreement with the trends detected in the experimental values and explain the conspicuous variations of the beta values primarily in terms of coupling between direct and resonant photoemission by interaction terms of different sign for different final vibrational states. PMID- 15836039 TI - Photoinduced processes in protonated tryptamine. AB - The electronic excited state dynamics of protonated tryptamine ions generated by an electrospray source have been studied by means of photoinduced dissociation technique on the femtosecond time scale. The result is that the initially excited state decays very quickly within 250 fs. The photoinduced dissociation channels observed can be sorted in two groups of fragments coming from two competing primary processes on the singlet electronic surface. The first one corresponds to a hydrogen-atom loss channel that creates a tryptamine radical cation. The radical cation subsequently fragments to smaller ions. The second process is internal conversion due to the H-atom recombination on the electronic ground state. Time-dependent density functional theory calculations show that an excited pisigma* state dissociative along the protonated amino N-H stretch crosses both the locally excited pipi* state and the electronic ground state S(0) and thus triggers the photofragmentation reactions. The two processes have equivalent quantum yields, approximately equal to 50% of the fragments coming from the H atom loss reaction. The two primary reaction paths can clearly be distinguished by their femtosecond pump/probe dynamics recorded on the different fragmentation channels. PMID- 15836040 TI - The binding of 4He and 3He to a hydrogen molecule: a computational study for pH2 and oH2. AB - One bound state for the very weakly interacting complex of H(2) with one He atom is computed using a recently obtained, very accurate potential energy surface [A. I. Boothroyd and P. G. Martin, J. Chem. Phys. 119, 3187 (2003)] and employing a discrete variable representation expansion. The spatial features of this very diffused, "halo" state are analyzed and discussed for various isotopic variants of the title system. PMID- 15836041 TI - Twisted intramolecular charge transfer states: rotationally resolved fluorescence excitation spectra of 4,4'-dimethylaminobenzonitrile in a molecular beam. AB - We report the observation at high resolution of seven vibronic bands that appear within approximately 200 cm(-1) of the electronic origin in the S(1)-S(0) fluorescence excitation spectrum of 4,4'-dimethylaminobenzonitrile (DMABN) in a molecular beam. Surprisingly, each band is found to be split into two or more components by a (coordinated) methyl group tunneling motion which significantly complicates the analysis. Despite this fact, high quality [(Observed-Calculated)< or =30 MHz] fits of each of the bands have been obtained, from which the rotational constants, inertial defects, torsion-rotation interaction constants, methyl group torsional barriers, and transition moment orientations of DMABN in both electronic states have been determined. The data show that DMABN is a slightly pyramidalized (approximately 1 degree) but otherwise (heavy-atom) planar molecule in its ground S(0) state, and that its electronically excited S(1) state has both a more pyramidalized (approximately 3 degrees) and twisted (approximately 25 degrees) dimethylamino group. Large reductions in the methyl group torsional barriers also show that the S(1)<--S(0) electronic transition is accompanied by significant charge transfer from the nitrogen atom to the pi* orbitals of the aromatic ring. Thereby established is the participation of all three vibrational coordinates in the dynamics leading to the "anomalous" emissive behavior of DMABN in the condensed phase. PMID- 15836042 TI - On the stability of glycine-water clusters with excess electron: implications for photoelectron spectroscopy. AB - Calculations are presented for the glycine-(H(2)O)(n) (-) (n=0-2) anionic clusters with excess electron, with the glycine core in the canonical or zwitterion form. A variety of conformers are predicted, and their relative energy is examined to estimate thermodynamic stability. The dynamic (proton transfer) pathways between the anionic clusters with the canonical and the zwitterion glycine core are examined. Small barrier heights for isomerization from the zwitterion glycine-(H(2)O)(2) (-) anion to those with canonical glycine core suggest that the former conformers may be kinetically unstable and unfavorable for detection of neutral glycine zwitterion-(H(2)O)(n) (n=1,2) clusters by photodetachment, in accordance with the photoelectron spectroscopic experiments by Bowen and co-workers [Xu et al., J. Chem. Phys. 119, 10696 (2003)]. The calculated stability of the glycine-(H(2)O)(n) (-) anion clusters with canonical glycine core relative to those with zwitterion core indicates that the observation of the anionic conformers with the canonical glycine core would be much more feasible, as revealed by Johnson and co-workers [Diken et al. J. Chem. Phys. 120, 9902 (2004)]. PMID- 15836043 TI - A density-functional study of the structures, binding energies and total spins of Ni-Fe clusters using nonlocal norm-conserving pseudopotentials and the generalized gradient approximation. AB - We report ab initio calculations of the structures, binding energies, and total spins of the clusters Ni(13), Ni(19), Ni(23), Ni(26), Ni(12)Fe, Ni(11)Fe(2), Ni(18)Fe, Ni(17)Fe(2), Ni(22)Fe, Ni(20)Fe(3), and Ni(25)Fe using a density functional method that employs linear combination of atomic orbitals as basis sets, nonlocal norm-conserving pseudopotentials, and the generalized gradient approximation for exchange and correlation. Our results show that the Fe-doped Ni clusters, which have icosahedral or polyicosahedral ground-state structures similar to those of the corresponding pure Ni clusters, are most stable with the Fe atoms occupying internal positions, as has also been inferred from experimental results on the adsorption of molecular nitrogen on the cluster surfaces. We also rule out the possibility that the experimentally observed difference between the (nonpolyicosahedral) configurations of N(2)-saturated Ni(26) and N(2)-saturated Ni(25)Fe be due to the influence of the Fe atom on the energy of the underlying metal cluster. PMID- 15836044 TI - The interpretation of molecular magnetic hyperfine interactions. AB - Investigations of the hyperfine structure in the excited electronic states of several free radical species have revealed shortcomings in the currently accepted values used for the theoretical interpretation of such interactions. We introduce updated reference atomic values from a combination of experimental observations and ab initio calculations. The latter are at Hartree-Fock and multireference configuration interaction levels of theory and several atomic test cases are discussed. Furthermore, ground and excited electronic state hyperfine coupling constants are calculated using both levels of theory for a range of first- and second-row diatomic hydride and nonhydride radicals. These results, together with a selection of other experimental measurements are then compared with experimental data where available, and the implications of the revised interpretation are discussed. PMID- 15836045 TI - A complete configurational study for the bound states of Ne trimers. AB - The structural properties and the energetics of the ground and the excited bound states of Ne(3) for zero total angular momentum are examined using different modelings for the two-body interactions. We employ a method consisting of a variational approach with a distributed Gaussian functions (DGF) basis set expansion. We discuss at length the advantages and possible limitations of such an approach, comparing it to other methods which have been applied in the literature to the same system. The DGF method turns out to be very accurate in giving us the bound states energetics and also provides in a natural way a convincing pictorial description of all the states, including those with dominant linear configurations. Additional bound states are found for the Ne(3) system with respect to those indicated in previous works and we suggest a "stabilization" procedure that can be used to assess the truly bound nature of a state. Some considerations on the relative reliability of the examined two-body interactions are also reported. PMID- 15836046 TI - Time-dependent density functional theory study on intramolecular charge transfer and solvent effect of dimethylaminobenzophenone. AB - The lower singlet excited states for dimethylaminobenzophenone have been investigated as a function of the twisting motion with inclusion of solvent effects. Theoretical calculations have been performed using time-dependent density functional theory. The B3LYP and MPW1PW91 functionals with a 6 311+G(2d,p) basis set have been used to compute transition energies. The solvent effects have been described within the polarizable continuum model. Ground-state geometries are optimized using density functional theory with both B3LYP and MPW1PW91 functionals combined with 6-31G(d) basis sets. Vertical absorption energy calculations characterize the lower singlet excited states both in vacuum and in different kinds of solvents. A large redshift of the absorption maximum in the polar solvent suggests an intramolecular charge transfer character of the excited state. We have constructed the potential energy curves of two possible twisting motions of the excited states both in vacuum and in the polar solvent of acetonitrile: the twisting of only the dimethylamino group and the twisting of the dimethylaminophenyl group with respect to the benzoyl group. Both twisting processes predict the formation of the twisted intramolecular charge transfer state associated with the crossing of a low barrier. The presence of the polar solvent significantly changes the shape of the energy curves. Calculated emission energies for both the isolated and the solvated systems show a large Stokes shift between the absorption and fluorescence maxima. Two possible twisting motions produce similar fluorescence spectroscopic consequences. Our results including solvent effects explain the weak "dual-fluorescence" feature of dimethylaminobenzophenone, and imply that the two possible twisting motions may occur in the excited-state relaxation dynamics, but the twisting of the dimethylamino group seems to take place easier. PMID- 15836047 TI - Reaction pathway for the nonadiabatic reaction of Ca(4s3d 1D)+H2-->CaH(X 2Sigma+)+H. AB - The reaction pathway and the nascent CaH product distribution in the reaction Ca(4s3d (1)D)+H(2)-->CaH(X (2)Sigma(+))+H are obtained using a pump-probe technique. The Ca atom is first prepared in the 3 (1)D state by a two-photon absorption, and then in brief time delay the laser-induced fluorescence of the reaction product CaH is monitored. The CaH(v=0,1) distributions appear to be single peaked, as characterized by Boltzmann rotational temperature of 807+/-38 K (v=0) and 684+/-77 K (v=1). The vibrational population ratio of CaH(v=0)/CaH(v=1) is determined to be 3.3+/-0.1, while the v=2 population is not detectable. The fractions of the available energy partitioning into rotation, vibration, and translation are estimated to be 0.36+/-0.05, 0.28+/-0.04, and 0.36+/-0.05, respectively. With the aid of the potential energy surfaces calculations, the current reaction should favor a near C(2v) collision configuration. The temperature dependence measurement yields a positive slope, indicative of the reaction occurrence without any potential barrier. The colliding species are anticipated to follow an attractive 1B(2) (or 2A') surface and then transit nonadiabatically to the reactive ground state surface. PMID- 15836048 TI - Statistical and nonstatistical nonadiabatic photodissociation from the first excited state of the hydroxymethyl radical. AB - Photodissociation of hydroxymethyl (H(2)COD) from the first excited state, a Rydberg 3s state, can produce either H+HCOD or H(2)CO+D. These processes involve a seam of conical intersections. An analysis of the seam is reported based on a pointwise determination of the three gradient vectors that characterize a conical intersection: the energy difference gradient and the coupling gradient, which span the branching space, and the gradient of the average energy. These data are used to understand why H, but not D is produced at energies near threshold, the difference in the recoil anisotropy for H and D dissociation, and whether H or D will be produced impulsively or after formation of an intermediate complex. PMID- 15836049 TI - Spectroscopy and metastability of CO2 2+ molecular ions. AB - The spectroscopy and metastability of the carbon dioxide doubly charged ion, the CO(2) (2+) dication, have been studied with photoionization experiments: time-of flight photoelectron photoelectron coincidence (TOF-PEPECO), threshold photoelectrons coincidence (TPEsCO), and threshold photoelectrons and ion coincidence (TPEsCO ion coincidence) spectroscopies. Vibrational structure is observed in TOF-PEPECO and TPEsCO spectra of the ground and first two excited states. The vibrational structure is dominated by the symmetric stretch except in the TPEsCO spectrum of the ground state where an antisymmetric stretch progression is observed. All three vibrational frequencies are deduced for the ground state and symmetric stretch and bending frequencies are deduced for the first two excited states. Some vibrational structure of higher electronic states is also observed. The threshold for double ionization of carbon dioxide is reported as 37.340+/-0.010 eV. The fragmentation of energy selected CO(2) (2+) ions has been investigated with TPEsCO ion coincidence spectroscopy. A band of metastable states from approximately 38.7 to approximately 41 eV above the ground state of neutral CO(2) has been observed in the experimental time window of approximately 0.1-2.3 mus with a tendency towards shorter lifetimes at higher energies. It is proposed that the metastability is due to slow spin forbidden conversion from bound excited singlet states to unbound continuum states of the triplet ground state. Another result of this investigation is the observation of CO(+)+O(+) formation in indirect dissociative double photoionization below the threshold for formation of CO(2) (2+). The threshold for CO(+)+O(+) formation is found to be 35.56+/-0.10 eV or lower, which is more than 2 eV lower than previous measurements. PMID- 15836050 TI - Cavity ring-down spectroscopy and theoretical calculations of the S1(1B3u)<- S0(1Ag) transition of jet-cooled perylene. AB - As part of our long-term program to test the diffuse interstellar band-polycyclic aromatic hydrocarbon hypothesis, we have investigated the S(1)<--S(0) electronic transition of neutral perylene (C(20)H(12)) in a combined experimental and theoretical study. Jet-cooled perylene was prepared with a pulsed discharge slit nozzle and detected by cavity ring-down spectroscopy. A number of vibronic features were observed in the 24 000-24 900 cm(-1) spectral range. Density functional and ab initio calculations were performed to determine the geometries, harmonic vibrational frequencies, and normal coordinates of both the S(0) and S(1) electronic states. A rotational temperature of 52+/-5 K was derived from a rotational contour analysis of the vibronic band associated with the 0-0 transition. A Franck-Condon treatment was carried out to calculate the vibronic spectrum of the S(1)<--S(0) transition. A good agreement was found between the calculated and the experimental spectra. A vibrational assignment is proposed and six normal modes are identified. The contribution of neutral compact polycyclic aromatic hydrocarbons to the diffuse interstellar bands is briefly discussed. PMID- 15836051 TI - Improved dissociation energy of the 39K85Rb molecule. AB - The predissociation data for the 1 (1)Pi state of (39)K(85)Rb of Kasahara et al. [J. Chem. Phys. 111, 8857 (1999)] are combined with the recent determination of the long range C(6) coefficients of the predissociating 2 (3)Sigma(+) approximately 2(0(-)), 2(1) states [Wang et al., Eur. Phys. J. D31, 165 (2004) ] molecule: to infer a more precise dissociation energy of the (39)K(85)Rb molecule D(0)=4180.06+/-0.42 cm(-1) and D(e)=4217.91+/-0.42 cm(-1). PMID- 15836052 TI - High resolution infrared spectra of H2-Kr and D2-Kr van der Waals complexes. AB - Infrared spectra of weakly bound hydrogen-krypton complexes have been studied at high spectral resolution (0.04 cm(-1)) using a long-path (154 m) low temperature (100 K) absorption cell and a Fourier transform spectrometer. In addition to spectra from the regions of the H(2) and D(2) fundamental vibrational bands in the midinfrared, the results also include the region of the pure rotational S(0)(0) transition of H(2) in the far infrared. A total of 219 measured line positions from these spectra have been fully assigned to specific quantum transitions and form the basis for determining a greatly improved semiempirical three-dimensional intermolecular potential energy surface for hydrogen-krypton in an accompanying paper. PMID- 15836053 TI - A reliable new three-dimensional potential energy surface for H(2)-Kr. AB - An improved three-dimensional potential energy surface for the H(2)-Kr system is determined from a direct fit of new infrared spectroscopic data for H(2)-Kr and D(2)-Kr to a potential energy function form based on the exchange-Coulomb model for the intermolecular interaction energy. These fits require repetitive, highly accurate simulations of the observed spectra, and both the strength of the potential energy anisotropy and the accuracy of the new data make the "secular equation perturbation theory" method used in previous analyses of H(2)-(rare gas) spectra inadequate for the present work. To address this problem, an extended version of the "iterative secular equation" method was developed which implements direct Hellmann-Feynman theorem calculation of the partial derivatives of eigenvalues with respect to parameters of the Hamiltonian which are required for the fits. PMID- 15836054 TI - 1H to 2H uniform cross-polarization nuclear magnetic resonance using 2H Lee Goldburg irradiation in static powders. AB - Application of conventional cross polarization (CP) to (2)H results in only a narrowband enhancement of the powder line shape due to the quadrupole interaction. We propose a CP scheme to uniformly enhance (2)H spectra in static powders. In this method, a Hartmann-Hahn matched (2)H rf field is applied on the Lee-Goldburg (LG) condition to remove the zeroth-order quadrupole interaction. In order to achieve a uniformly enhanced (2)H powder line shape with a limited (2)H rf intensity, the (1)H rf amplitude in CP is stepwise altered during the contact time. We develop a spin-thermodynamic theory to describe polarization transfer due to CP with LG irradiation (LG-CP) under the influence of the quadrupole interaction, which can successfully reproduce the LG-CP line shapes observed under various experimental conditions. Experimental and simulated (2)H powder spectra are reported for some compounds. PMID- 15836055 TI - An ab initio study of the lowest electronic states of yttrium dicarbide, YC2. AB - The low-lying electronic states of yttrium dicarbide have been calculated using highly correlated wave functions and systematic sequences of correlation consistent basis sets. For the (2)A(1) ground electronic state, the near equilibrium potential energy surface (PES) has been calculated using the coupled cluster method in conjunction with basis sets ranging in size from double to quintuple zeta. The relativistic effects have been taken into account by using pseudopotentials for the Y atom. After extrapolation to the complete basis set limit, additional corrections due to core-valence correlation and spin-orbit effects have also been included. The same approach has been followed for the (2)B(1), (2)B(2), and (2)A(2) states but only the C(2V) PESs have been considered in these cases. For the two (2)A(1) electronic excited states and, for comparison purposes, for the ground state, the multireference configuration interaction (MRCI) approach has been used in conjunction with double-zeta and triple-zeta basis sets for the construction of the PES. The molecular and spectroscopic properties predicted for the ground and excited states investigated in this work compare well with the available experimental data, particularly for the ground electronic state. The 0 K dissociation enthalpy of YC(2), DeltaH(Y-C2)(0 K), and its atomization enthalpy, SigmaD(0), are predicted to be 148.4 and 291.5 kcal/mol, respectively. PMID- 15836056 TI - Role of surface crossings in the photochemistry of nitromethane. AB - The photodissociation dynamics of nitromethane (CH(3)NO(2)) starting at the S(3) excited state has been studied at the complete active space self-consistent field level of theory in conjunction with atomic natural orbital type basis sets. In addition, the energies of all the critical points and the energy profiles connecting them have been recomputed with the multiconfigurational second-order perturbation method. It is found that the key step in the reaction mechanism is a radiationless decay through an S(3)S(2) conical intersection. The branching space spanned by the gradient difference and nonadiabatic coupling vectors of this crossing point comprises dissociation into excited nitromethane plus singlet atomic oxygen [CH(3)NO(1A")+O((1)D)] and S(3)-->S(2) deactivation, respectively. Furthermore, deactivated nitromethane S(n (n<3)) can decompose in subsequent steps into CH(3)+NO(2), where NO(2) is generated at least in two different electronic states (1 (2)B(2) and 1 (2)A(1)). It is shown that formation of excited nitric oxide NO(A (2)Sigma) arises from CH(3)NO(1A") generated in the previous step. In addition, four crossings between singlet and triplet states are localized; however, no evidence is found for a relevant role of such crossings in the photochemistry of CH(3)NO(2) initiated at S(3) state in the gas phase. PMID- 15836057 TI - The shear viscosity of supercritical oxygen at high pressure. AB - Shear viscosities of supercritical oxygen have been measured up to a pressure of 5.7 GPa at 294 K. A modified free-volume expression fits the data within 6% between the limits of the tenuous gas and 4.8 times the critical density. Nitrogen viscosities were found to correspond to those of oxygen through a simple scaling by critical constants. Viscosities were measured in the high-pressure diamond-anvil cell with a rolling-ball technique. The dynamics of a sphere rolling on an inclined plane were investigated in the context of these experiments. The effect of a second surface, situated above the sphere, was experimentally determined. PMID- 15836058 TI - Coherence spectroscopy in dissipative media: a Liouville space pathway approach. AB - We address the possibility of using coherent control tools to extract useful information about the interaction of a system with a dissipative environment. To that end we extend previous work, which developed a coherence spectroscopy based on two-pathway excitation phase control, from the isolated molecule limit to dense media. Specifically, we explore the properties of the channel phase, an observable of energy-domain two-pathway excitation experiments that was shown in the isolated molecule limit to carry information about the phase properties of the material system. Our analysis is based on the combination of steady state and time-dependent analytical perturbative approaches within the density matrix formalism, complemented by nonperturbative numerical simulations. We find that the channel phase carries significantly richer information in the presence of decoherence mechanisms than in their absence. In particular, rescattering events in the structured continuum introduce new features in the channel phase spectrum, whose structure conveys information about both the molecular continuum and the system bath interaction. PMID- 15836059 TI - Modified Kelvin-Thomson equation considering ion-dipole interaction: comparison with observed ion-clustering enthalpies and entropies. AB - The classical Kelvin-Thomson (CKT) equation does not consider the interaction of condensing molecules with the ions and hence is not able to treat polar and nonpolar molecules differently. The ion-clustering enthalpy and entropy changes predicted by CKT equation for small ions are known to be significantly less negative than those observed. In this paper, we derive a modified Kelvin-Thomson (MKT) equation, which considers the effect of dipole-ion interaction, by taking into account the kinetic energy change of condensing polar ligands as they approach the ions or the extra energy needed for dipole molecules to escape from the ion cluster. The clustering enthalpies and entropies for protonated clusters (H(+)L(n), with L=H(2)O, NH(3), CH(3)OH, and C(5)H(5)N) are calculated based on MKT equation and compared with experimental data. Our calculations indicate that enthalpy values given by MKT equation are in very good agreement with experimental results for small ions (n< or =5) of all four species investigated. MKT predictions appear to be consistent with observed enthalpies for CH(3)OH at n> or =6 and for H(2)O at n=14-25, however, MKT equation cannot reproduce the observed discontinuous transition in enthalpy changes at n=6 for NH(3) and at n=7 13 for H(2)O which is probably associated with the formation of inner shell. The stepwise entropy changes for small ions are 5-15 cal mol(-1) K(-1) more negative when the effect of dipole-ion interaction is considered, which suggests that the ordered structure of the cluster ions can somewhat be accounted for by the dipole ion interaction term. PMID- 15836060 TI - Temperature dependence of the double layer capacitance for the restricted primitive model of an electrolyte solution from a density functional approach. AB - We apply a different version of the density functional theory, given by Pizio, Patrykiejew, and Sokolowski [J. Chem. Phys. 121, 11957 (2004)], for a nonuniform restricted primitive model of an electrolyte solution to evaluate the temperature dependence of the capacitance of an electric double layer. We show that this theory is capable of reproducing the computer simulation data at a quantitative level. In particular, the reversal of the temperature dependence of the capacitance at low temperatures is predicted. This phenomenon has been difficult to predict from theory. Further, this theory also leads to an accurate description of the double layer structure. PMID- 15836061 TI - Dynamics of water in a molecular sieve by quasielastic neutron scattering. AB - We have investigated the dynamics of water confined in a molecular sieve, with a cylindrical pore diameter of 10 A, by means of quasielastic neutron scattering (QENS). Both the incoherent and coherent intermediate scattering functions I(Q,t) were determined by time-of-flight QENS and the neutron spin-echo technique, respectively. The results show that I(Q,t) is considerably more stretched in time with a slightly larger average relaxation time in the case of coherent scattering. From the Q dependence of I(Q,t) it is clear that the observed dynamics is almost of an ordinary translational nature. A comparison with previous dielectric measurements suggests a possible merging of the alpha and beta relaxations of the confined water at T=185 K, although the alpha relaxation cannot be directly observed at lower temperatures due to the severe confinement. The present results are discussed in relation to previous results for water confined in a Na-vermiculite clay, where the average relaxation time from spin echo measurements was found to be slower than in the present system (particularly at low temperatures). PMID- 15836062 TI - Ultrafast tabletop laser-pump-x-ray probe measurement of solvated Fe(CN)6 4-. AB - We report on the first ultrafast laser-pump-picosecond x-ray probe measurements of solvated transition metal complexes carried out with a tabletop ultrafast laser-driven plasma x-ray source. The x-ray absorption fine structure (XAFS) spectra of Fe(CN)(6) (4-) solvated in water have been measured before and tens of picoseconds after photoexcitation with ultrashort UV laser pulses. The XAFS spectra after photoexcitation exhibits a K-edge shift indicating the increase of the iron-ligand distances. Reference spectra of Fe(CN)(6) (4-) and Fe(CN)(6) (3-) measured at a synchrotron source yield structural data that show static solvation induced bond length changes of the metal complexes. PMID- 15836063 TI - A phenomenological model of dynamical arrest of electron transfer in solvents in the glass-transition region. AB - A phenomenological model of electron transfer reactions in solvents undergoing glass transition is discussed. The reaction constant cuts off slow polarization modes from the spectrum of nuclear thermal motions active on the observation time scale. The arrest of nuclear solvation in turn affects the reaction activation barrier making it dependent on the rate. The resultant rate constant is sought from a self-consistent equation. The model describes well the sharp change in the solvent Stokes shift of optical lines in the glass-transition region. It is also applied to describe the temperature dependence of primary charge separation and reduction of primary pair in photosynthetic reaction centers. The model shows that a weak dependence of the primary charge separation rate on temperature can be explained by dynamical arrest of nuclear solvation on the picosecond time scale of electron transfer. For reduction of primary pair by cytochrome, the model yields a sharp turnover of the reaction kinetics at the transition temperature when nuclear solvation freezes in. PMID- 15836064 TI - Dynamics of glassy and liquid m-toluidine investigated by high-resolution dielectric spectroscopy. AB - The glass-former m-toluidine displays the characteristic properties of a fragile supercooled liquid, which suggest the existence of a slow secondary relaxation process. In view of the recently realized importance of such a secondary relaxation feature, we have conducted a dielectric search for the secondary process in viscous and glassy m-toluidine. Based on high-resolution experiments on the distilled liquid, a secondary process can be identified which has the properties typical of a Johari-Goldstein beta relaxation. As a result, the previous hypothesis that the methyl group might be responsible for suppressing the secondary dynamics in glassy m-toluidine no longer holds. PMID- 15836065 TI - Non-quadratic-intensity dependence of two-photon absorption induced fluorescence of organic chromophores in solution. AB - We have provided a model to interpret the non-quadratic-intensity dependence behavior commonly observed in the two-photon fluorescence (TPF) experiment excited with high laser intensity. The model also provides one with a different technique to measure the two-photon absorption cross section of an organic chromophore in solution. In contrast to the commonly used low intensity technique that depends on the quadratic-intensity law, the present technique is based on the non-quadratic-intensity dependence of two-photon fluorescence. Auxiliary data such as two-photon quantum efficiency and fluorescence collection efficiency, essential in the low intensity method, are not required in the present technique. TPF measurements of Rhodamine B in methanol are carried out to demonstrate the validity of the present method. The method is used to determine the two-photon absorption cross section of a new chromophore attached with tricyano-derivatized furan as the electron acceptor. The two-photon absorption cross section measured using this method is also compared with that using a conventional transmission technique. PMID- 15836066 TI - Pair correlation function of short-ranged square-well fluids. AB - We have performed extensive Monte Carlo simulations in the canonical (NVT) ensemble of the pair correlation function for square-well fluids with well widths lambda-1 ranging from 0.1 to 1.0, in units of the diameter sigma of the particles. For each one of these widths, several densities rho and temperatures T in the ranges 0.1< or =rhosigma(3)< or =0.8 and T(c)(lambda) less or approximately T less or approximately 3T(c)(lambda), where T(c)(lambda) is the critical temperature, have been considered. The simulation data are used to examine the performance of two analytical theories in predicting the structure of these fluids: the perturbation theory proposed by Tang and Lu [Y. Tang and B. C. Y. Lu, J. Chem. Phys. 100, 3079 (1994); 100, 6665 (1994)] and the nonperturbative model proposed by two of us [S. B. Yuste and A. Santos, J. Chem. Phys. 101 2355 (1994)]. It is observed that both theories complement each other, as the latter theory works well for short ranges and/or moderate densities, while the former theory works for long ranges and high densities. PMID- 15836067 TI - Primary and secondary relaxations in bis-5-hydroxypentylphthalate. AB - Broadband dielectric spectroscopy was used to study the relaxation dynamics in bis-5-hydroxypentylphthalate (BHPP) under both isobaric and isothermal conditions. The relaxation dynamics exhibit complex behavior, arising from hydrogen bonding in the BHPP. At ambient pressure above the glass transition temperature T(g), the dielectric spectrum shows a broad structural relaxation peak with a prominent excess wing toward higher frequencies. As temperature is decreased below T(g), the excess wing transforms into two distinct peaks, both having Arrhenius behavior with activation energies equal to 58.8 and 32.6 kJmol for slower (beta) and faster (gamma) processes, respectively. Furthermore, the relaxation times for the beta process increase with increasing pressure, whereas the faster gamma relaxation is practically insensitive to pressure changes. Analysis of the properties of these secondary relaxations suggests that the beta peak can be identified as an intermolecular Johari-Goldstein (JG) process. However, its separation in frequency from the alpha relaxation, and both its activation energy and activation volume, differ substantially from values calculated from the breadth of the structural relaxation peak. Thus, the dynamics of BHPP appear to be an exception to the usual correlation between the respective properties of the structural and the JG secondary relaxations. PMID- 15836068 TI - Kinetics and mechanism of bimolecular electron transfer reaction in quinone-amine systems in micellar solution. AB - Photoinduced electron transfer (ET) reactions between anthraquinone derivatives and aromatic amines have been investigated in sodium dodecyl sulphate (SDS) micellar solutions. Significant static quenching of the quinone fluorescence due to high amine concentration in the micellar phase has been observed in steady state measurements. The bimolecular rate constants for the dynamic quenching in the present systems k(q) (TR), as estimated from the time-resolved measurements, have been correlated with the free energy changes DeltaG(0) for the ET reactions. Interestingly it is seen that the k(q) (TR) vs DeltaG(0) plot displays an inversion behavior with maximum k(q) (TR) at around 0.7 eV, a trend similar to that predicted in Marcus ET theory. Like the present results, Marcus inversion in the k(q) (TR) values was also observed earlier in coumarin-amine systems in SDS and TX-100 micellar solutions, with maximum k(q) (TR) at around the same exergonicity. These results thus suggest that Marcus inversion in bimolecular ET reaction is a general phenomenon in micellar media. Present observations have been rationalized on the basis of the two-dimensional ET (2DET) theory, which seems to be more suitable for micellar ET reactions than the conventional ET theory. For the quinone-amine systems, it is interestingly seen that k(q) (TR) vs DeltaG(0) plot is somewhat wider in comparison to that of the coumarin-amine systems, even though the maxima in the k(q) (TR) vs DeltaG(0) plots appear at almost similar exergonicity for both the acceptor-donor systems. These observations have been rationalized on the basis of the differences in the reaction windows along the solvation axis, as envisaged within the framework of the 2DET theory, and arise due to the differences in the locations of the quinones and coumarin dyes in the micellar phase. PMID- 15836069 TI - Solvent mediated interactions close to fluid-fluid phase separation: microscopic treatment of bridging in a soft-core fluid. AB - Using density functional theory we calculate the density profiles of a binary solvent adsorbed around a pair of big solute particles. All species interact via repulsive Gaussian potentials. The solvent exhibits fluid-fluid phase separation, and for thermodynamic states near to coexistence the big particles can be surrounded by a thick adsorbed "wetting" film of the coexisting solvent phase. On reducing the separation between the two big particles we find there can be a "bridging" transition as the wetting films join to form a fluid bridge. The effective (solvent mediated) potential between the two big particles becomes long ranged and strongly attractive in the bridged configuration. Within our mean field treatment the bridging transition results in a discontinuity in the solvent mediated force. We demonstrate that accounting for the phenomenon of bridging requires the presence of a nonzero bridge function in the correlations between the solute particles when our model fluid is described within a full mixture theory based upon the Ornstein-Zernike equations. PMID- 15836070 TI - Hydration of Li+ -ion in atom-bond electronegativity equalization method-7P water: a molecular dynamics simulation study. AB - We have carried out molecular dynamics simulations of a Li(+) ion in water over a wide range of temperature (from 248 to 368 K). The simulations make use of the atom-bond electronegativity equalization method-7P water model, a seven-site flexible model with fluctuating charges, which has accurately reproduced many bulk water properties. The recently constructed Li(+)-water interaction potential through fitting to the experimental and ab initio gas-phase binding energies and to the measured structures for Li(+)-water clusters is adopted in the simulations. ABEEM was proposed and developed in terms of partitioning the electron density into atom and bond regions and using the electronegativity equalization method (EEM) and the density functional theory (DFT). Based on a combination of the atom-bond electronegativity equalization method and molecular mechanics (ABEEM/MM), a new set of water-water and Li(+)-water potentials, successfully applied to ionic clusters Li(+)(H(2)O)(n)(n=1-6,8), are further investigated in an aqueous solution of Li(+) in the present paper. Two points must be emphasized in the simulations: first, the model allows for the charges on the interacting sites fluctuating as a function of time; second, the ABEEM-7P model has applied the parameter k(lp,H)(R(lp,H)) to explicitly describe the short range interaction of hydrogen bond in the hydrogen bond interaction region, and has a new description for the hydrogen bond. The static, dynamic, and thermodynamic properties have been studied in detail. In addition, at different temperatures, the structural properties such as radial distribution functions, and the dynamical properties such as diffusion coefficients and residence times of the water molecules in the first hydration shell of Li(+), are also simulated well. These simulation results show that the ABEEM/MM-based water-water and Li(+) water potentials appear to be robust giving the overall characteristic hydration properties in excellent agreement with experiments and other molecular dynamics simulations on similar system. PMID- 15836071 TI - Structure and dynamics of electrostatically interacting magnetic nanoparticles in suspension. AB - We investigate the structure and dynamics of charge-stabilized CoFe(2)O(4)-SiO(2) core-shell magnetic nanoparticles in suspensions. Small angle x-ray scattering and x-ray photon correlation spectroscopy allow us to analyze the intraparticle (core-shell) and interparticle structure of the suspension, as well as their dynamic and hydrodynamic behavior. Due to the weak magnetic interactions, the liquidlike structure is governed by screened Coulomb interactions. The hydrodynamic interactions of the measured systems are significantly stronger than predicted by current theories. PMID- 15836072 TI - Electrophoresis of ionic microgel particles: from charged hard spheres to polyelectrolyte-like behavior. AB - We perform electrophoretic mobility measurements of ionic microgel particles in the deswollen and swollen phases. The results show that microgels behave as charged hard spheres in the first case and as free-draining spherical polyelectrolytes in the latter. A unified theory for the electrophoresis of polyelectrolyte-coated particles [H. Ohshima, Adv. Colloid Interface Sci. 62, 189 (1995)] is shown to contain the essential physics for describing the experiments, upon adequate consideration of the particles swelling behavior and network solvent friction variations. PMID- 15836073 TI - The stress diffusion coupling in the swelling dynamics of cylindrical gels. AB - The swelling dynamics of long cylindrical gel is analyzed by the stress diffusion coupling model [T. Yamaue and M. Doi, Phys. Rev. E 69, 41402 (2004)]. Two situations are analyzed: (i) stress induced swelling, where the swelling is caused by an elongational force applied on the gel, and (ii) free swelling, where the swelling is caused by thermodynamic force. The relaxation times characterizing these processes are calculated. It is shown that earlier calculations for the relaxation time, which are based on some physical or mathematical approximations, give results surprisingly close to the rigorous calculation, but the difference can be still seen experimentally. PMID- 15836074 TI - Nucleation in a simple model for protein solutions with anisotropic interactions. AB - A lattice analog of density functional theory is used to explore the structural and thermodynamic properties of critical nuclei in mixtures of particles with attractive anisotropic interactions. Protein molecules are assumed to occupy the sites on a regular cubic lattice, with effective directional interactions that mimic hydrogen bonding and the solvation forces induced by water. Interaction parameters are chosen to qualitatively reproduce the phase behavior of protein solutions. Our model predicts that critical nuclei of the solidlike phase have nonspherical shapes, and that their specific geometry depends on the nature of the anisotropic interactions. Molecules tend to align in distinctive ways in the core and in the interfacial region of these critical clusters, and the width and structure of the interface are highly affected by the presence of a metastable fluid-fluid critical point. Close to the critical region, the height of the barrier to nucleation is strongly reduced; this effect is enhanced by increasing the anisotropy of the intermolecular interactions. Unlike systems with short range isotropic interactions, nucleation in our model is initiated by highly ordered clusters in which the order-disorder transition is confined to the interfacial region. PMID- 15836075 TI - Anomalous corresponding-states surface tension of hydrogen fluoride and of the Onsager model. AB - In a corresponding-states analysis of the liquid-vapor surface tension originally suggested by Guggenheim, we study the behavior of different simple (i.e., nonpolar), polar and ionic fluids. The results are compared to the corresponding ones for model fluids of each of the three types. For simple and weakly polar fluids (both real and model), the data map onto a master curve, as demonstrated by Guggenheim. For strongly dipolar, associating fluids, which also exhibit hydrogen bonding, one finds deviations from the master curve at low temperatures and, thus, observes the characteristic sigmoid behavior of the reduced surface tension as a function of temperature. The same is obtained for the model ionic fluid, the restricted primitive model. Truly exceptionally low values of the reduced surface tension are found for hydrogen fluoride and for the Onsager model of dipolar fluids, the surface tension of which we evaluate using an approximate hypernetted chain relation to obtain the square-gradient term in a modified van der Waals theory. Remarkably, in the corresponding-states plot, the surface tensions of HF and of the Onsager model agree very closely, while being well separated from the values for the other fluids. We also study the gradual transition of a model fluid from a simple fluid to a strongly dipolar one by varying the relative strength of dipolar and dispersion forces. PMID- 15836076 TI - Effects of triplet exciton confinement induced by reduced conjugation length in polyspirobifluorene copolymers. AB - Using gated optical spectroscopy at low temperatures, a polyspirobifluorene has been compared with an alternating carbazole-spirobifluorene copolymer in which the backbone conjugation is interrupted due to meta coupling of the carbazole moieties. In the copolymer both singlet and triplet energy levels are blueshifted by 130 meV with respect to the homopolymer, resulting in an unaltered singlet-to triplet splitting. Though the barrier for triplet exciton migration increases from 4.4 to 6.0 meV for the copolymer compared to the homopolymer, it still remained low enough to ensure efficient triplet diffusion at ambient temperature. PMID- 15836077 TI - Conformational transition free energy profiles of an adsorbed, lattice model protein by multicanonical Monte Carlo simulation. AB - Proteins often undergo changes in internal conformation upon interacting with a surface. We investigate the thermodynamics of surface induced conformational change in a lattice model protein using a multicanonical Monte Carlo method. The protein is a linear heteropolymer of 27 segments (of types A and B) confined to a cubic lattice. The segmental order and nearest neighbor contact energies are chosen to yield, in the absence of an adsorbing surface, a unique 3x3x3 folded structure. The surface is a plane of sites interacting either equally with A and B segments (equal affinity surface) or more strongly with the A segments (A affinity surface). We use a multicanonical Monte Carlo algorithm, with configuration bias and jump walking moves, featuring an iteratively updated sampling function that converges to the reciprocal of the density of states 1/Omega(E), E being the potential energy. We find inflection points in the configurational entropy, S(E)=k ln Omega(E), for all but a strongly adsorbing equal affinity surface, indicating the presence of free energy barriers to transition. When protein-surface interactions are weak, the free energy profiles F(E)=E-TS(E) qualitatively resemble those of a protein in the absence of a surface: a free energy barrier separates a folded, lowest energy state from globular, higher energy states. The surface acts in this case to stabilize the globular states relative to the folded state. When the protein surface interactions are stronger, the situation differs markedly: the folded state no longer occurs at the lowest energy and free energy barriers may be absent altogether. PMID- 15836078 TI - Distribution patterns and controllable transport of water inside and outside charged single-walled carbon nanotubes. AB - The density distribution patterns of water inside and outside neutral and charged single-walled carbon nanotubes (SWNTs) soaked in water have been studied using molecular dynamics simulations based on TIP3P potential and Lennard-Jones parameters of CHARMM force field, in conjunction with ab initio calculations to provide the electron density distributions of the systems. Water molecules show different electropism near positively and negatively charged SWNTs. Different density distribution patterns of water, depending on the diameter and chirality of the SWNTs, are observed inside and outside the tube wall. These special distribution patterns formed can be explained in terms of the van der Waals and electrostatic interactions between the water molecules and the carbon atoms on the hexagonal network of carbon nanotubes. The electric field produced by the highly charged SWNTs leads to high filling speed of water molecules, while it prevents them from flowing out of the nanotube. Water molecules enter the neutral SWNTs slowly and can flow out of the nanotube in a fluctuating manner. It indicates that by adjusting the electric charge on the SWNTs, one can control the adsorption and transport behavior of polar molecules in SWNTs to be used as stable storage medium with template effect or transport channels. The transport rate can be tailored by changing the charge on the SWNTs. PMID- 15836079 TI - A density functional theory study of sulfur poisoning. AB - Density functional theory calculations have been used to investigate the chemisorption of H, S, SH, and H(2)S as well as the hydrogenation reactions S+H and SH+H on a Rh surface with steps, Rh(211), aiming to explain sulfur poisoning effect. In the S hydrogenation from S to H(2)S, the transition state of the first step S+H-->SH is reached when the S moves to the step-bridge and H is on the off top site. In the second step, SH+H-->H(2)S, the transition state is reached when SH moves to the top site and H is close to another top site nearby. Our results show that it is difficult to hydrogenate S and they poison defects such as steps. In order to address why S is poisoning, hydrogenation of C, N, and O on Rh(211) has also been calculated and has been found that the reverse and forward reactions possess similar barriers in contrast to the S hydrogenation. The physical origin of these differences has been analyzed and discussed. PMID- 15836080 TI - Electronic nonadiabatic effects in the adsorption of hydrogen atoms on metals. AB - The time-dependent, mean-field Newns-Anderson model for a spin-polarized adsorbate approaching a metallic surface is solved in the wide-band limit. Equations for the time evolution of the occupation of the spin dependent adsorbate states and for the nonadiabatic and nearly adiabatic adsorbate-surface energy transfer rates are derived. Numerical solutions are obtained using characteristic parameters derived from density functional theory calculations for the H/Cu(111) system. The time evolution of the model system is shown to be strongly nonadiabatic in the vicinity of the transition point between spin polarized and nonpolarized ground states. Away from the spin transition the nonadiabatic energy transfer is in close agreement with the nearly adiabatic limit. Near the transition, nonadiabatic effects are large and the nearly adiabatic approximation fails. PMID- 15836081 TI - Kinetic theory of gas separation in a nanopore and comparison to molecular dynamics simulation. AB - Kinetic mesoscopic theory derived from an atomistic model is applied to study permeation and separation of gases in a single rectangular pore. The goal is to judge the analytical method against the results of molecular dynamics simulation and to demonstrate the ease and relevance of analytical theories to calculate density profiles, flux, permeance, and separation factors. The permeance is linked to the amount of gas adsorbed in the pore and the effect of the effective gas-wall interaction on adsorption is explored. The effects of pore size, temperature, and the parameters of the pore wall interaction are investigated and reproduce the trends found in the numerical simulation of permeation of a mixture of methane and carbon dioxide in a carbon nanopore. PMID- 15836082 TI - Characterization of the interface dipole at the paraphenylenediamine-nickel interface: a joint theoretical and experimental study. AB - In organic-based (opto)electronic devices, charge injection into conjugated materials is governed to a large extent by the metal-organic interface dipole. Controlling the injection of charges requires a better understanding of the fundamental origin of the interface dipole. In this context, photoelectron spectroscopies and density functional theory calculations are used to investigate the interaction between para-phenylenediamine (PPDA), an electron donor, and a polycrystalline nickel surface. The interface dipole formed upon chemisorption of one PPDA monolayer strongly modifies the work function of the nickel surface from 5.10 to 3.55 eV. The work function decrease of 1.55 eV is explained by the electron-donor character of PPDA and the modification of the electronic density at the metal surface. PPDA monolayers are composed of tilted molecules interacting via the nitrogen lone-pair and PPDA molecules chemisorbed parallel to the surface via their pi-electron density. Annealing the monolayer leads to dehydrogenation of PPDA activated by the nickel surface, as found for other amines. PMID- 15836083 TI - Local reaction rates and surface diffusion on nanolithographically prepared model catalysts: experiments and simulations. AB - Combining molecular beam methods and angular resolved mass spectrometry, we have studied the angular distribution of desorbing products during CO oxidation on a planar Pd/silica supported model catalyst. The model catalyst was prepared by means of electron beam lithography, allowing individual control of particle size, position, and aspect ratio, and was characterized by atomic force microscopy and scanning electron microscopy before and after reaction. In the experiment, both oxygen and CO rich regimes were investigated using separate molecular beams for the two reactants. This allows exploration of diffusion effects of reactants on the particles and of shadowing and backscattering phenomena. A reaction-diffusion model was developed in order to extract information about local reaction rates on the surface of the catalyst nanoparticles. The model takes into account the structural parameters of the catalyst as well as the backscattering of the reactants and products from the support. It allows a quantitative description of the experimental data and provides a detailed understanding of temperature and reactant flux dependent effects. Moreover, information on the surface mobility of oxygen under steady-state reaction conditions could be obtained by comparison with the experimental results. PMID- 15836084 TI - How many dimensions are required to approximate the potential energy landscape of a model protein? AB - A scheme to approximate the multidimensional potential energy landscape in terms of a minimal number of degrees of freedom is proposed using a linear transformation of the original atomic Cartesian coordinates. For one particular off-lattice model protein the inherent frustration can only be reproduced satisfactorily when a relatively large number of coordinates are employed. However, when this frustration is removed in a Go-type model, the number of coordinates required is significantly lower, especially around the global potential energy minimum. To aid our interpretation of the results we consider modified disconnectivity graphs where a measure of the structural diversity and a metric relation between the stationary points are incorporated. PMID- 15836085 TI - Growth-induced polarity formation in solid solutions of organic molecules: Markov mean-field model and Monte Carlo simulations. AB - A layer-by-layer growth model is presented for the theoretical investigation of growth-induced polarity formation in solid solutions H(1-X)G(X) of polar (H) and nonpolar (G) molecules (X: molar fraction of G molecules in the solid, 0 or =0.8) fractions of G molecules, respectively, as well as for ordered HG compounds (X=0.5). PMID- 15836086 TI - Origin of temperature dependent conductivity of alpha-polyvinylidene fluoride. AB - The conductivity of alpha-polyvinylidene fluoride is obtained from dielectric measurements performed in the frequency domain at several temperatures. At temperatures above the glass-transition, the conductivity can be interpreted as an ionic conductivity, which confirms earlier results reported in the literature. Our investigation shows that the observed ionic conductivity is closely related to the amorphous phase of the polymer. PMID- 15836087 TI - Stoichiometric polyelectrolyte complexes as comb copolymers. AB - The collapse behavior of a single comblike copolymer chain has been studied by Monte Carlo simulations. It has been supposed that the solvent is good for the side chains but the solvent quality for the backbone chain changes. It has been shown that depending on the structural parameters of the comb copolymer (the lengths of the backbone and side chains, grafting density of the side chains) various thermodynamically stable morphologies of the collapsed backbone chain can be realized. In addition to ordinary spherical globule we have observed elongated structures as well as necklace-like conformations. The proposed model can be used to describe conformational behavior of stoichiometric complexes between block copolymers with a polyelectrolyte short block and oppositely charged linear homopolymers. PMID- 15836088 TI - Brownian dynamics algorithm for bead-rod semiflexible chain with anisotropic friction. AB - A model of semiflexible bead-rod chain with anisotropic friction can mimic closely the hydrodynamics of a slender filament. We present an efficient algorithm for Brownian dynamics simulations of this model with configuration dependent anisotropic bead friction coefficients. The algorithm is an extension of that given previously for the case of configuration independent isotropic friction coefficients by Grassia and Hinch [J. Fluid Mech. 308, 255 (1996)]. We confirm that the algorithm yields predicted values for various equilibrium properties. We also present a stochastic algorithm for evaluation of the stress tensor, and we show that in the limit of stiff chains the algorithm recovers the results of Kirkwood and Plock [J. Chem. Phys. 24, 665 (1956)] for rigid rods with hydrodynamic interactions. PMID- 15836089 TI - Surface enrichment of branched polymers in linear hosts: effect of asymmetry in intersegmental interactions and density gradients. AB - Variable density lattice treatment of surface enrichment of f-arm star-branched chains in star/linear polymer blends is compared with results of an analytical response theory proposed by Wu and Fredrickson [Macromolecules 29, 7919 (1996)]. We find that differences in treating the intersegmental interactions in the small interfacial region near a free surface lead to significant differences in the potentials by which polymer chain ends are attracted towards the surface. Consideration of an asymmetric relationship between segment potentials and density changes in polystyrene at 450 K and 0.1 MPa, for example, gives typically a threefold to fourfold enhancement in composition of star molecules at a vacuum interface. When contributions from gradients in density are included in the analysis even greater levels of surface enhancement (fivefold to sixfold increases) are observed. By appropriately estimating the attraction of chain ends and repulsion of branch points at a free surface, we show that concentration profiles of branched polymers predicted in the lattice model are consistent with results obtained in the analytical response theory. PMID- 15836090 TI - The size of a polymer in a symmetric solvent. AB - Using a simple thermodynamic model, we derive an expression for the excluded volume parameter v of a polymer chain in a symmetric solvent (solvated by its own monomers). For a chain with a given segment length and number of monomers, this parameter determines whether the chain is collapsed or expanded. For the latter it determines the degree of expansion. Using a simple off-lattice version of Flory's model [P. J. Flory, Principles of Polymer Chemistry (Cornell University Press, Ithaca, 1953)] and relaxing the assumption of incompressibility, we obtain the result v=(1-kappa)rho(0), where kappa is the dimensionless compressibility and rho(0) the number density of solvent. In the incompressible limit (in the sense that kappa-->0) the chain is expanded and the inverse of the solvent number density determines the degree of expansion of the chain. Using the van der Waals equation of state to estimate kappa (allowing for nonzero compressibility in a system that can undergo a gas-liquid phase transition), the model predicts that upon raising the temperature at constant pressure there is both a lower (coil to globule) and upper (globule to coil) Flory temperature. This is in quantitative agreement with experiment and computer simulations. PMID- 15836091 TI - Computer simulation study of intermolecular voids in unsaturated phosphatidylcholine lipid bilayers. AB - Computer simulation of the liquid crystalline phase of five different hydrated unsaturated phosphadidylcholine (PC) lipid bilayers, i.e., membranes built up by 18:0/18:1omega9cis PC, 18:0/18:2omega6cis PC, 18:0/18:3omega3cis PC, 18:0/20:4omega6cis PC, and 18:0/22:6omega3cis PC molecules have been performed on the isothermal-isobaric ensemble at 1 atm and 303 K. (The notation n:domegapcis specifies the lipid tails: n refers to the total number of carbon atoms in the chain, d is the number of the methylene-interrupted double bonds, p denotes the number of carbons between the chain terminal CH(3) group and the nearest double bond, and cis refers to the conformation around the double bonds.) The characteristics of the free volume in these systems have been analyzed by means of a generalized version of the Voronoi-Delaunay method [M. G. Alinchenko et al., J. Phys. Chem. B 108, 19056 (2004)]. As a reference system, the hydrated bilayer of the saturated 14:014:0 PC molecules (dimyristoylphosphatidylcholine) has also been analyzed. It has been found that the profiles of the fraction of the free volume across the membrane exhibit a rather complex pattern. This fine structure of the free volume fraction profiles can be interpreted by dividing the membrane into three separate major zones (i.e., zones of the aqueous, polar, and apolar parts of the membrane) and defining five subzones within these zones according to the average position of various atomic groups in the membrane. The fraction of the free volume in the middle of the membrane is found to increase with increasing unsaturation of the sn-2 chain of the lipid molecule. This is due to the fact that with increasing number of methylene-interrupted double bonds the lipid tails become more flexible, and hence they do not extend to the middle of the membrane. It is found that there are no broad enough preformed channels in the bilayers through which small penetrants, such as water molecules, can readily go through; however, the existing channels can largely facilitate the permeation of these molecules. PMID- 15836092 TI - Chain length dependence of the thermodynamic properties of linear and cyclic alkanes and polymers. AB - The specific heat capacity was measured with step-scan differential scanning calorimetry for linear alkanes from pentane (C(5)H(12)) to nonadecane (C(19)H(40)), for several cyclic alkanes, for linear and cyclic polyethylenes, and for a linear and a cyclic polystyrene. For the linear alkanes, the specific heat capacity in the equilibrium liquid state decreases as chain length increases; above a carbon number N of 10 (decane) the specific heat asymptotes to a constant value. For the cyclic alkanes, the heat capacity in the equilibrium liquid state is lower than that of the corresponding linear chains and increases with increasing chain length. At high enough molecular weights, the heat capacities of cyclic and linear molecules are expected to be equal, and this is found to be the case for the polyethylenes and polystyrenes studied. In addition, the thermal properties of the solid-liquid and the solid-solid transitions are examined for the linear and cyclic alkanes; solid-solid transitions are observed only in the odd-numbered alkanes. The thermal expansion coefficients and the specific volumes of the linear and cyclic alkanes are also calculated from literature data and compared with the trends in the specific heats. PMID- 15836093 TI - Threading-unthreading equilibrium in solution of molecular nanotubes and linear flexible polymer chains. AB - Using the Flory-Huggins lattice model, we investigate the threading-unthreading equilibrium in a solution of linear flexible polymer chains and molecular nanotubes formed by covalently bonding the ringlike molecules, such as cyclodextrins (CDs). It is found that the threading-unthreading equilibrium depends on the temperature and the molar concentrations of the ringlike molecules and polymer chain segments but is independent of the polymer chain length, which agrees with the experimental observations. By fitting the experimental data of alpha-CD and poly(ethylene glycol) (PEG), the inclusion energy between alpha-CD and PEG, which includes the conformation energy loss of PEG resulted from the inclusion, is calculated to be approximately -20.45 kJ/mol per PEG unit. PMID- 15836094 TI - Comment on "Excitations in photoactive molecules from quantum Monte Carlo" [J. Chem. Phys. 121, 5836 (2004)]. AB - It is shown that the qualitative differences between high-level ab initio calculations and restricted open-shell Kohn-Sham (ROKS) results for the lowest singlet excited electronic state of formaldimine along a particular isomerization path found by Schautz, Buda, and Filippi [J. Chem. Phys.121, 5836 (2004)] play a minor role in molecular dynamics simulations of photoisomerization at room temperature. In fact, ROKS yields, within its well-known limitations, a good representation of the physically relevant isomerization pathway. PMID- 15836096 TI - Comment on "First principles electrochemistry: electrons and protons reacting as independent ions" [J. Chem. Phys. 117, 10193 (2002)]. AB - The entropy of the electron gas used by Llano and Eriksson (LE) in the definition they used for an absolute half-cell potential (AHCP) is indeterminate. This AHCP is therefore not based on a thermodynamically well-defined process and cannot easily be extended to other thermodynamic functions such as entropy. An alternative approach to the AHCP avoids these difficulties. The present comment also corrects statements and concepts presented by LE about the temperature coefficient of the Fermi level and surface charge of the electrolyte solution. PMID- 15836098 TI - Comment on "Radiation damage of biosystems mediated by secondary electrons: Resonant precursors for uracil molecules" [J. Chem. Phys. 120, 7446 (2004)]. AB - Resonances computed by Gianturco and Lucchese are incorrectly correlated with those observed experimentally. A shift of their pi* resonances by 2 eV gives satisfactory agreement. Arguments for determining whether a resonance is a suitable precursor for the fragmentation products are too simplistic, and in one example, incorrect. PMID- 15836101 TI - Continuous configuration time-dependent self-consistent field method for polyatomic quantum dynamical problems. AB - A new continuous configuration time-dependent self-consistent field method has been developed to study polyatomic dynamical problems by using the discrete variable representation for the reaction system, and applied to a reaction system coupled to a bath. The method is very efficient because the equations involved are as simple as those in the traditional single configuration approach, and can account for the correlations between the reaction system and bath modes rather well. PMID- 15836102 TI - Accelerating quantum mechanical/molecular mechanical sampling using pure molecular mechanical potential as an importance function: the case of effective fragment potential. AB - Acceleration of sampling from a quantum mechanical/effective fragment mechanical (QM/EFP) potential is explored with effective fragment potential (EFP) as an importance function. EFP, generated on the fly, is found to be an excellent choice for an importance function for a QM/EFP potential. This technique is used to find nine stationary points of a blocked amino acid with twelve waters in a semi-automated way. PMID- 15836103 TI - The stabilization of arginine's zwitterion by dipole-binding of an excess electron. AB - The arginine parent anion was generated by a newly developed, infrared desorption electron photoemission hybrid anion source. The photoelectron spectrum of the arginine anion was recorded and interpreted as being due to dipole binding of the excess electron. The results are consistent with calculations by Rak, Skurski, Simons, and Gutowski, who predicted the near degeneracy of arginine's canonical and zwitterionic dipole bound anions. Since neutral arginine's zwitterion is slightly less stable than its canonical form, this work also demonstrates the ability of an excess electron to stabilize a zwitterion, just as ions and solvent molecules are already known to do. PMID- 15836104 TI - Novel method for selective probing of ground-state rotational dynamics of solutes in solvents. AB - We introduce an optical pump/probe method that allows selective measurement of ground-state rotational dynamics of solutes in liquids. It relies on employing two successive pump pulses that are adjusted to create an optical anisotropy due to the orientational distribution of only the ground-state solute molecules. Measurement on a dye-solvent system shows a large difference between the rotational diffusion rates of the ground state and the excited state of the dye molecules due to different solute-solvent interactions. PMID- 15836105 TI - Isomer selective infrared spectroscopy of neutral metal clusters. AB - We report experimental infrared spectra of neutral metal clusters in the gas phase. Multiple photon dissociation of the argon complexes of niobium clusters is used to obtain vibrational spectra in the 80-400 cm(-1) region. The observed spectra for Nb(9)Ar(n) (n=1-4) are different for different values of n. This is explained by the presence of two isomers of Nb(9) that have different affinities towards Ar and the isomer specific infrared spectra are obtained. The structures of the isomers are determined by comparing the observed spectra with the outcome of density-functional theory calculations. PMID- 15836106 TI - Molecular wave packet interferometry and quantum entanglement. AB - We study wave packet interferometry (WPI) considering the laser pulse fields both classical and quantum mechanically. WPI occurs in a molecule after subjecting it to the interaction with a sequence of phase-locked ultrashort laser pulses. Typically, the measured quantity is the fluorescence of the molecule from an excited electronic state. This signal has imprinted the interference of the vibrational wave packets prepared by the different laser pulses of the sequence. The consideration of the pulses as quantum entities in the analysis allows us to study the entanglement of the laser pulse states with the molecular states. With a simple model for the molecular system, plus several justified approximations, we solve for the fully quantum mechanical molecule-electromagnetic field state. We then study the reduced density matrices of the molecule and the laser pulses separately. We calculate measurable corrections to the case where the fields are treated classically. PMID- 15836107 TI - Mixed quantum-classical equilibrium. AB - We present an analysis of the equilibrium limits of the two most widely used approaches for simulating the dynamics of molecular systems that combine both quantum and classical degrees of freedom. For a two-level quantum system connected to an infinite number of classical particles, we derive a simple analytical expression for the equilibrium mean energy attained by the self consistent-field (Ehrenfest) method and show that it deviates substantially from Boltzmann. By contrast, "fewest switches" surface hopping achieves Boltzmann quantum state populations. We verify these analytical results with simulations. PMID- 15836108 TI - Dynamics of coupled Bohmian and phase-space variables: a moment approach to mixed quantum-classical dynamics. AB - The theoretical framework of the mixed quantum-classical description given by Burghardt and Parlant [J. Chem. Phys. 120, 3055 (2004)] is detailed. A representation in terms of partial hydrodynamic moments is developed, the dynamics of which is determined by a hierarchy of equations derived from the quantum Liouville equation. Exact equations of motion are obtained, whose quantum classical approximants are associated with a fluid-dynamical trajectory representation which couples classical variables to quantum hydrodynamic variables. The latter evolve under a generalized hydrodynamic force which also depends upon the classical phase-space variables. The hydrodynamic moment description is shown to be closely connected to mixed quantum-classical phase space methods. PMID- 15836109 TI - Numerical study of the accuracy and efficiency of various approaches for Monte Carlo surface hopping calculations. AB - A one-dimensional, two-state model problem with two well-separated avoided crossing points is employed to test the efficiency and accuracy of a semiclassical surface hopping technique. The use of a one-dimensional model allows for the accurate numerical evaluation of both fully quantum-mechanical and semiclassical transition probabilities. The calculations demonstrate that the surface hopping procedure employed accounts for the interference between different hopping trajectories very well and provides highly accurate transition probabilities. It is, in general, not computationally feasible to completely sum over all hopping trajectories in the semiclassical calculations for multidimensional problems. In this case, a Monte Carlo procedure for selecting important trajectories can be employed. However, the cancellation due to the different phases associated with different trajectories limits the accuracy and efficiency of the Monte Carlo procedure. Various approaches for improving the accuracy and efficiency of Monte Carlo surface hopping procedures are investigated. These methods are found to significantly reduce the statistical sampling errors in the calculations, thereby increasing the accuracy of the transition probabilities obtained with a fixed number of trajectories sampled. PMID- 15836110 TI - Third- and fourth-order perturbation corrections to excitation energies from configuration interaction singles. AB - Complete third-order and partial fourth-order Rayleigh-Schrodinger perturbation corrections to excitation energies from configuration interaction singles (CIS) have been derived and termed CIS(3) and CIS(4)(P). They have been implemented by the automated system TENSOR CONTRACTION ENGINE into parallel-execution programs taking advantage of spin, spatial, and index permutation symmetries and applicable to closed- and open-shell molecules. The consistent use of factorization, first introduced by Head-Gordon et al. in the second-order correction to CIS denoted CIS(D), has reduced the computational cost of CIS(3) and CIS(4)(P) from O(n(8)) and O(n(6)) to O(n(6)) and O(n(5)), respectively, with n being the number of orbitals. It has also guaranteed the size extensivity of excited-state energies of these methods, which are in turn the sum of size intensive excitation energies and the ground-state energies from the standard Moller-Plesset perturbation theory at the respective orders. The series CIS(D), CIS(3), and CIS(4)(P) are usually monotonically convergent at values close to the accurate results predicted by coupled-cluster singles and doubles (CCSD) with a small fraction of computational costs of CCSD for predominantly singly excited states characterized by a 90%-100% overlap between the CIS and CCSD wave functions. When the overlap is smaller, the perturbation theory is incapable of adequately accounting for the mixing of the CIS states through higher-than singles sectors of the Hamiltonian matrix, resulting in wildly oscillating series with often very large errors in CIS(4)(P). Hence, CIS(3) and CIS(4)(P) have a rather small radius of convergence and a limited range of applicability, but within that range they can be an inexpensive alternative to CCSD. PMID- 15836111 TI - Role of angular momentum conservation in unimolecular translational energy release: validity of the orbiting transition state theory. AB - The translational kinetic energy release distribution (KERD) for the halogen loss reaction of the bromobenzene and iodobenzene cations has been reinvestigated on the microsecond time scale. Two necessary conditions of validity of the orbiting transition state theory (OTST) for the calculation of kinetic energy release distributions (KERDs) have been formulated. One of them examines the central ion induced dipole potential approximation. As a second criterion, an adiabatic parameter is derived. The lower the released translational energy and the total angular momentum, the larger the reduced mass, the rotational constant of the molecular fragment, and the polarizability of the released atom, the more valid is the OTST. Only the low-energy dissociation of the iodobenzene ion (E approximately 0.45 eV, where E is the internal energy above the reaction threshold) is found to fulfill the criteria of validity of the OTST. The constraints that act on the dissociation dynamics have been studied by the maximum entropy method. Calculations of entropy deficiencies (which measure the deviation from a microcanonical distribution) show that the pair of fragments does not sample the whole of the phase space that is compatible with the mere specification of the internal energy. The major constraint that results from conservation of angular momentum is related to a reduction of the dimensionality of the dynamics of the translational motion to a two-dimensional space. A second and minor constraint that affects the KERD leads to a suppression of small translational releases, i.e., accounts for threshold behavior. At high internal energies, the effects of curvature of the reaction path and of angular momentum conservation are intricately intermeddled and it is not possible to specify the share of each effect. PMID- 15836112 TI - Application of variational reduced-density-matrix theory to organic molecules. AB - Variational calculation of the two-electron reduced-density matrix (2-RDM), using a new first-order algorithm [D. A. Mazziotti, Phys. Rev. Lett. 93, 213001 (2004)], is applied to medium-sized organic molecules. The calculations reveal systematic trends in the accuracy of the energy with well-known chemical concepts such as hybridization, electronegativity, and atomic size. Furthermore, correlation energies from hydrocarbon chains indicate that the calculation of the 2-RDM subject to two-positivity conditions is size extensive, that is, the energy grows linearly with the number of electrons. Because organic molecules have a well-defined set of functional groups, we employ the trends in energy accuracy of the functional groups to design a correction to the 2-RDM energy for an arbitrary organic molecule. We apply the 2-RDM calculations with the functional-group correction to a large set of organic molecules with different functional groups. Energies with millihartree accuracy are obtained both at equilibrium and nonequilibrium geometries. PMID- 15836113 TI - Stationary phase evaluations of quantum rate constants. AB - We compute the quantum rate constant based on two extended stationary phase approximations to the imaginary-time formulation of the quantum rate theory. The optimized stationary phase approximation to the imaginary-time flux-flux correlation function employs the optimized quadratic reference system to overcome the inaccuracy of the quadratic expansion in the standard stationary phase approximation, and yields favorable agreements with instanton results for both adiabatic and nonadiabatic processes in dissipative and nondissipative systems. The integrated stationary phase approximation to the two-dimensional barrier free energy is particularly useful for adiabatic processes and demonstrates consistent results with the imaginary-time flux-flux correlation function approach. Our stationary phase methods do not require calculation of tunneling paths or stability matrices, and work equally well in the high-temperature and the low temperature regimes. The numerical results suggest their general applicability for calibration of imaginary-time methods and for the calculation of quantum rate constants in systems with a large number of degrees of freedom. PMID- 15836114 TI - Testing wave packet dynamics in computing radiative association cross sections. AB - A time-dependent wave packet method is used to compute cross sections for radiative recombination reactions using the Li((2)S)+H(+)-->LiH(+)(X (2)Sigma(+))+gamma as a test case. Cross sections are calculated through standard time-to-energy mapping of the time-dependent transition moment and a useful method is introduced to deal with the low collision energy regime. Results are in quantitative agreement over the whole energy range 10(-4)-5 eV with previous time independent results for the same system [I. Baccarelli, L. Andric, T. Grozdanov, and R. McCarroll, J. Chem. Phys. 117, 3013 (2002)], thereby suggesting that the method can be of help in computing radiative association cross sections for more complicated systems. PMID- 15836115 TI - Electron correlation in Hooke's law atom in the high-density limit. AB - Closed-form expressions for the first three terms in the perturbation expansion of the exact energy and Hartree-Fock energy of the lowest singlet and triplet states of the Hooke's law atom are found. These yield elementary formulas for the exact correlation energies (-49.7028 and -5.807 65 mE(h)) of the two states in the high-density limit and lead to a pair of necessary conditions on the exact correlation kernel G(w) in Hartree-Fock-Wigner theory. PMID- 15836116 TI - Multireference configuration interaction based electronic Floquet states for molecules in an intense radiation field: theory and application to Li(2)+. AB - A multireference configuration interaction (CI) method which includes single and double excitations based description of adiabatic Floquet states for the electronic structure of a molecule in an intense laser field is introduced. Using a variant of a recently introduced configuration state function (CSF) based Table CI methodology, it is shown that the multiple states of several irreducible representations required for a good description of low-lying Floquet states can be obtained using modifications of computational molecular electronic structure techniques. In particular, formulas for all components of the transition dipole moment matrix elements within the CSF-based Table-CI method are derived and presented. Moreover, the flexibility of the recently introduced macroconfiguration description of model and external configuration spaces is shown to lead to multiple potential energy surfaces of sufficiently uniform quality to allow construction of useful Floquet states. The formalism and computer programs developed are demonstrated on Li(2) (+) in a 0.9 x 10(12) W/cm(2) field. In analogy with Na(2) (+), the 1,2 (2)Sigma(g) (+), 1,2 (2)Sigma(u) (+), 1 (2)Pi(g), and 1 (2)Pi(u) states are of relevance, although the pattern of couplings is shown to be more complex. A hitherto unnoticed metastable state, which correlates asymptotically with 2 (2)Sigma(u) (+), is described. PMID- 15836117 TI - Ab initio calculation of the C/D ratio of magnetic circular dichroism. AB - A procedure for calculating the magnetic circular dichroism C/D ratio from density functional theory calculations is discussed. The method is simplified considerably through the application of group theory and the irreducible-tensor method and only requires integrals of the magnetic dipole moment operator over a few orbitals and published tables of symmetry factors. The implementation of the method is tested through application to several small and medium-sized molecules. PMID- 15836118 TI - Local-MP2 electron correlation method for nonconducting crystals. AB - Rigorous methods for the post-HF (HF-Hartree-Fock) determination of correlation corrections for crystalline solids are currently being developed following different strategies. The CRYSTAL program developed in Torino and Daresbury provides accurate HF solutions for periodic systems in a basis set of Gaussian type functions; for insulators, the occupied HF manifold can be represented as an antisymmetrized product of well localized Wannier functions. This makes possible the extension to nonconducting crystals of local correlation linear scaling On techniques as successfully and efficiently implemented in Stuttgart's MOLPRO program. These methods exploit the fact that dynamic electron correlation effects between remote parts of a molecule (manifesting as dispersive interactions in intermolecular perturbation theory) decay as an inverse sixth power of the distance R between these fragments, that is, much more quickly than the Coulomb interactions that are treated already at the HF level. Translational symmetry then permits the crystalline problem to be reduced to one concerning a cluster around the reference zero cell. A periodic local correlation program (CRYSCOR) has been prepared along these lines, limited for the moment to the solution of second-order Moller-Plesset equations. Exploitation of point group symmetry is shown to be more important and useful than in the molecular case. The computational strategy adopted and preliminary results concerning five semiconductors with tetrahedral structure (C, Si, SiC, BN, and BeS) are presented and discussed. PMID- 15836119 TI - Operator splitting algorithm for isokinetic SLLOD molecular dynamics. AB - We apply an operator splitting method to develop a simulation algorithm that has complete analytical solutions for the Gaussian thermostated SLLOD equations of motion [D. J. Evans and G. P. Morriss, Phys. Rev. A 30, 1528 (1984)] for a system under shear. This leads to a homogeneous algorithm for performing both equilibrium and nonequilibrium isokinetic molecular dynamics simulation. The resulting algorithm is computationally efficient. In particular, larger integration time steps can be used compared to simulations with regular Gaussian thermostated SLLOD equations of motion. The utility and accuracy of the algorithm are demonstrated through application to the Weeks-Chandler-Anderson fluid. Although strict conservation of the kinetic energy suppresses thermal fluctuations in the system, this algorithm does not allow simulations at lower shear rates than those normally afforded by older nonequilibrium molecular dynamics simulations. PMID- 15836120 TI - The merits of the frozen-density embedding scheme to model solvatochromic shifts. AB - We investigate the usefulness of a frozen-density embedding scheme within density functional theory [J. Phys. Chem. 97, 8050 (1993)] for the calculation of solvatochromic shifts. The frozen-density calculations, particularly of excitation energies have two clear advantages over the standard supermolecule calculations: (i) calculations for much larger systems are feasible, since the time-consuming time-dependent density functional theory (TDDFT) part is carried out in a limited molecular orbital space, while the effect of the surroundings is still included at a quantum mechanical level. This allows a large number of solvent molecules to be included and thus affords both specific and nonspecific solvent effects to be modeled. (ii) Only excitations of the system of interest, i.e., the selected embedded system, are calculated. This allows an easy analysis and interpretation of the results. In TDDFT calculations, it avoids unphysical results introduced by spurious mixings with the artificially too low charge transfer excitations which are an artifact of the adiabatic local-density approximation or generalized gradient approximation exchange-correlation kernels currently used. The performance of the frozen-density embedding method is tested for the well-studied solvatochromic properties of the n-->pi(*) excitation of acetone. Further enhancement of the efficiency is studied by constructing approximate solvent densities, e.g., from a superposition of densities of individual solvent molecules. This is demonstrated for systems with up to 802 atoms. To obtain a realistic modeling of the absorption spectra of solvated molecules, including the effect of the solvent motions, we combine the embedding scheme with classical molecular dynamics (MD) and Car-Parrinello MD simulations to obtain snapshots of the solute and its solvent environment, for which then excitation energies are calculated. The frozen-density embedding yields estimated solvent shifts in the range of 0.20-0.26 eV, in good agreement with experimental values of between 0.19 and 0.21 eV. PMID- 15836121 TI - Describing static correlation in bond dissociation by Kohn-Sham density functional theory. AB - We show that density functional theory within the RPA (random phase approximation for the exchange-correlation energy) provides a correct description of bond dissociation in H(2) in a spin-restricted Kohn-Sham formalism, i.e., without artificial symmetry breaking. We present accurate adiabatic connection curves both at equilibrium and beyond the Coulson-Fisher point. The strong curvature at large bond length implies important static (left-right) correlation, justifying modern hybrid functional constructions but also demonstrating their limitations. Although exact at infinite separation and accurate near the equilibrium bond length, the RPA dissociation curve displays unphysical repulsion at larger but finite bond lengths. Going beyond the RPA by including the exact exchange kernel (RPA+X), we find a similar repulsion. We argue that this deficiency is due to the absence of double excitations in adiabatic linear response theory. Further analyzing the H(2) dissociation limit we show that the RPA+X is not size consistent, in contrast to the RPA. PMID- 15836122 TI - Experiments and quantum-chemical calculations on Rydberg states of H2CS in the region 5.6-9.5 eV. AB - Absorption spectrum of H(2)CS in the region 5.6-9.5 eV was recorded with a continuously tunable light source of synchrotron radiation. After we subtracted absorption bands of CS(2), our spectrum clearly shows vibrational progressions associated with transitions (1)A(1)(pi,pi*)-X (1)A(1) and (1)B(2)(n,4s)-X (1)A(1) in the region 5.6-6.7 eV. A spectrum from which absorption of C(2)H(4) and CS(2) are subtracted shows several discrete bands in the region 6.9-9.5 eV. A Rydberg state (1)B(2)(n,4p(z)) lying below Rydberg state (1)A(1)(n,4p(y)) is confirmed, and the C-H symmetric stretching (nu(1)) and CH out-of-plane bending (nu(4)) modes for a transition (1)B(2)(n,4s)-X (1)A(1) are identified. New transitions to Rydberg states associated with excitation to 5s-11s, 5p(z)-7p(z), 5p(y)-7p(y), and 3d-6d are identified based on quantum defects and comparison with vertical excitation energies predicted with time-dependent density-functional theory (TD DFT) and outer-valence Green's-function (OVGF) methods. For lower excited states predictions from these TD-DFT6-31+G calculations agree satisfactorily with experimental values, but for higher Rydberg states the OVGF method using aug-cc pVTZ basis set augmented with extra diffuse functions yields more accurate predictions of excitation energies. PMID- 15836123 TI - Excited-state decay of hydrocarbon radicals, investigated by femtosecond time resolved photoionization: ethyl, propargyl, and benzyl. AB - The excited state decay of the hydrocarbon radicals ethyl, C(2)H(5); propargyl, C(3)H(3); and benzyl, C(7)H(7) was investigated by femtosecond time-resolved photoionization. Radicals were generated by flash pyrolysis of n-propyl nitrite, propargyl bromide, and toluene, respectively. It is shown that the 2 (2)A(') (3s) Rydberg state of ethyl excited at 250 nm decays with a time constant of 20 fs. No residual signal was observed at longer delay times. For the 3 (2)B(1) state of propargyl excited at 255 nm a slower decay with a time constant 50+/-10 fs was determined. The 4 (2)B(2) state of benzyl excited at 255 nm decays within 150+/ 30 fs. PMID- 15836124 TI - Structures of [(CO2)n(H2O)m]- (n=1-4, m=1,2) cluster anions. I. Infrared photodissociation spectroscopy. AB - The infrared photodissociation spectra of [(CO(2))(n)(H(2)O)(m)](-) (n=1-4, m=1, 2) are measured in the 3000-3800 cm(-1) range. The [(CO(2))(n)(H(2)O)(1)](-) spectra are characterized by a sharp band around 3570 cm(-1) except for n=1; [(CO(2))(1)(H(2)O)(1)](-) does not photodissociate in the spectral range studied. The [(CO(2))(n)(H(2)O)(2)](-) (n=1, 2) species have similar spectral features with a broadband at approximately 3340 cm(-1). A drastic change in the spectral features is observed for [(CO(2))(3)(H(2)O)(2)](-), where sharp bands appear at 3224, 3321, 3364, 3438, and 3572 cm(-1). Ab initio calculations are performed at the MP2/6-311++G(**) level to provide structural information such as optimized structures, stabilization energies, and vibrational frequencies of the [(CO(2))(n)(H(2)O)(m)](-) species. Comparison between the experimental and theoretical results reveals rather size- and composition-specific hydration manner in [(CO(2))(n)(H(2)O)(m)](-): (1) the incorporated H(2)O is bonded to either CO(2) (-) or C(2)O(4) (-) through two equivalent OH...O hydrogen bonds to form a ring structure in [(CO(2))(n)(H(2)O)(1)](-); (2) two H(2)O molecules are independently bound to the O atoms of CO(2) (-) in [(CO(2))(n)(H(2)O)(2)](-) (n=1, 2); (3) a cyclic structure composed of CO(2) (-) and two H(2)O molecules is formed in [(CO(2))(3)(H(2)O)(2)](-). PMID- 15836125 TI - The singlet electronic ground state isomers of dialuminum monoxide: AlOAl, AlAlO, and the transition state connecting them. AB - The singlet electronic ground state isomers, X (1)Sigma(g) (+) (AlOAl D(infinityh)) and X (1)Sigma(+) (AlAlO C(infinitynu)), of dialuminum monoxide have been systematically investigated using ab initio electronic structure theory. The equilibrium structures and physical properties for the two molecules have been predicted employing self-consistent field (SCF) configuration interaction with single and double excitations (CISD), multireference CISD (MRCISD), coupled cluster with single and double excitations (CCSD), CCSD with perturbative triples [CCSD(T)], CCSD with iterative partial triple excitations (CCSDT-3 and CC3), and full triples (CCSDT) coupled cluster methods. Four correlation consistent polarized valence (cc-pVXZ) type basis sets were used. The AlAlO system is rather challenging theoretically. The two isomers are confirmed to have linear structures at all levels of theory. The symmetric isomer AlOAl is predicted to lie 81.9 kcal mol(-1) below the asymmetric isomer AlAlO at the cc pV(Q+d)Z CCSD(T) level of theory. The predicted harmonic vibrational frequencies for the X (1)Sigma(g) (+) AlOAl molecule, omega(1)=517 cm(-1), omega(2)=95 cm( 1), and omega(3)=1014 cm(-1), are in good agreement with experimental values. The harmonic vibrational frequencies for the X (1)Sigma(+) AlAlO structure, omega(1)=1042 cm(-1), omega(2)=73 cm(-1), and omega(3)=253 cm(-1), presently have no experimental values with which to be compared. With the same methods the barrier heights for the isomerization AlOAl-->AlAlO and AlAlO-->AlOAl reactions were predicted to be 84.3 and 2.4 kcal mol(-1), respectively. The dissociation energies D(0) for AlOAl (X (1)Sigma(g) (+)) and AlAlO (X (1)Sigma(+))-->AlO (X (2)Sigma(+))+Al ((2)P) were determined to be 130.8 and 48.9 kcal mol(-1), respectively. Thus, both symmetric AlOAl (X (1)Sigma(g) (+)) and asymmetric AlAlO (X (1)Sigma(+)) isomers are expected to be thermodynamically stable with respect to the dissociation into AlO (X (2)Sigma(+)) + Al ((2)P) and kinetically stable for the isomerization reaction (AlAlO-->AlOAl) at sufficiently low temperatures. PMID- 15836126 TI - Electronic decay following ionization of aqueous Li+ microsolvation clusters. AB - Ionization in the energetical range between 35 eV and 75 eV of aqueous Li(+) microsolvation clusters may initialize several different electronic decay processes. Electronic decay following H(2)O 2s ionization in a cationic cluster is reported. Li ionization probes the efficiency of electron transfer mediated decay (ETMD) processes. We report estimated ETMD lifetimes in the range of 20-100 fs for clusters with one to five water monomers. Furthermore, tertiary electron emission may occur via a combined cascade of electron transfer mediated decay and intermolecular Coulombic decay. PMID- 15836127 TI - Revision of the second ionization energy of toluene. AB - Charge stripping (CS) of the molecular ion of toluene, C(7)H(8) (+)-->C(7)H(8) (2+)+e, is often used as a reference for the determination of second ionization energies in energy-resolved CS experiments. For calibration of the kinetic energy scale, a value of IE(C(7)H(8) (+))=(15.7+/-0.2) eV derived from the appearance energy of the toluene dication upon electron ionization has been accepted generally. Triggered by some recent discrepancies between CS measurements on the one hand and different experimental methods as well as theoretical predictions on the other, we have reinvestigated the photon-induced double ionization of toluene using synchrotron radiation. These photoionization measurements yield phenomenological appearance energies of AE(C(7)H(8) (+))=(8.81+/-0.03) eV for the monocation and AE(C(7)H(8) (2+))=(23.81+/-0.06) eV for the dication. The former is in good agreement with a much more precise spectroscopic value, IE(C(7)H(8))=(8.8276+/-0.0006) eV. Explicit consideration of the Franck-Condon envelopes associated with photoionization to the dication in conjunction with the application of the Wannier law leads to an adiabatic ionization energy IE(a)(C(7)H(8) (+))=(14.8+/-0.1) eV, which is as much as 0.9 eV lower than the previous value derived from electron ionization. Because in many previous CS measurements the transition C(7)H(8) (+)-->C(7)H(8) (2+)+e was used as a reference, the energetics of several gaseous dications might need some readjustment. PMID- 15836128 TI - Interaction of NH(X 3Sigma-) with He: potential energy surface, bound states, and collisional Zeeman relaxation. AB - A detailed analysis of the He-NH((3)Sigma(-)) van der Waals complex is presented. We discuss ab initio calculations of the potential energy surface and fitting procedures with relevance to cold collisions, and we present accurate calculations of bound energy levels of the triatomic complex as well as collisional properties of NH molecules in a buffer gas of (3)He. The influence of the external magnetic field used to trap the NH molecules and the effect of the atom-molecule interaction potential on the collisionally induced Zeeman relaxation are explored. It is shown that minute variations of the interaction potential due to different fitting procedures may alter the Zeeman relaxation rate at ultralow temperatures by as much as 50%. PMID- 15836129 TI - Parallel temperatures in supersonic beams: ultracooling of light atoms seeded in a heavier carrier gas. AB - Supersonic expansion is a very powerful tool to produce an atomic beam with a well defined velocity and, by seeding a test gas in such an expansion, the energy of the test gas can be transferred, at least partially, to the very-low temperature carrier gas. The case usually studied is the one of a heavy gas seeded in a light carrier gas and, in this case, the parallel temperature of the seeded gas is always larger than the one of the carrier gas. In the present paper, we study the opposite case which has received less attention: when a light gas is seeded in a heavier carrier gas, the parallel temperature can be substantially lower for the seeded gas than for the carrier gas. This effect has been first observed by Campargue and co-workers in 2000, in the case of atomic oxygen seeded in argon. In the present paper, we develop a theoretical analysis of this effect, in the high dilution limit, and we compare our theoretical results to several experimental observations, including a set of measurements we have made on a beam of lithium seeded in argon. The agreement between theory and experiments is good. PMID- 15836130 TI - A comparative study for elastic electron collisions on the isoelectronic CNN, NCN, and CCO radicals. AB - In this work, we present a theoretical study on elastic electron collisions from three isoelectronic free radicals (CNN, NCN, and CCO) in the low incident energy range. More specifically, calculated differential, integral, and momentum transfer cross sections are reported in the 1-30 eV energy range. Calculations are performed in the static-exchange and static-exchange-polarization levels. The iterative Schwinger variational method is used to solve the scattering equations. Our study reveals that the calculated cross sections for the three targets are significantly different at incident energies below 10 eV. Above that energy, a remarkable similarity among the calculated results is seen. PMID- 15836131 TI - Structure and energetics of small gold nanoclusters and their positive ions. AB - We performed density functional theory calculations at the PW91PW91/LANL2DZ, PW91PW91/Stuttgart 1997, PW91PW91/CRENBL, B3LYP/LANL2DZ, and SVWN5/LANL2DZ levels of theory to attain the minimum-energy structures of neutral and cationic gold clusters of up to nine gold atoms. We locate the 2D-to-3D (two-dimensional to three-dimensional) transition in cationic clusters as occurring between Au(8) (+) and Au(9) (+). We also demonstrate that we can correlate the 2D-to-3D transition in cationic clusters with a linear extrapolation of the energy differences of the lowest-lying 2D and 3D structures of cluster sizes below the transition. We then use the same approach to predict that the 2D-to-3D transition occurs in neutral clusters at Au(11); this is confirmed by locating 3D Au(11) structures that are lower in energy than the best 2D structures reported to date. We examine the effects of choice of basis sets and exchange-correlation functionals on the relative stabilities and other properties of the calculated structures. Finally we find that there is good agreement between calculated and experimental data for clusters with up to six constituent atoms. For clusters with more than seven atoms, there are significant differences observed between the calculated and experimental properties using SVWN5/LANL2DZ, but there is still good agreement for the other levels of theory used. PMID- 15836132 TI - Photoisomerization of azobenzene from first-principles constrained density functional calculations. AB - Despite considerable work in the field, the precise mechanism for the photoisomerization of azobenzene, C(12)H(10)N(2), is still an open issue. Early theoretical studies of the problem indicated that isomerization occurs through an in-plane inversion path, and this has been used to explain recent time-resolved UV-visible spectroscopy measurements. On the other hand, a number of recent theoretical studies have concluded that a torsion of the N-N bond ("rotation path") is probably the most favorable mechanism for photoisomerization involving the first excited state. We have performed first-principles calculations using constrained density-functional theory (DFT) and time-dependent DFT in the local density approximation, with results that also favor the rotation path mechanism. Our results are compared with other analyses, primarily based on configuration interaction. PMID- 15836133 TI - Anion and cation-yield spectroscopy of core-excited SF6. AB - We report an extensive study on total and partial-ion-yield spectroscopy around both the S 2p and F 1s thresholds in SF(6). All positive and negative single-ion channels have been measured. Below the F 1s threshold we detect a large variation in relative intensity of the resonant structures according to the specific channel monitored, indicating selective fragmentation. Above threshold, at variance with previous cases described by us, we detect high-intensity structures related to shape resonances not only in the cation channels but also for the anions. We discuss the applicability and limits of a model we have developed for the analysis of shape resonances in anion yields as a function of molecular size. PMID- 15836134 TI - Structures of Mo2Oy- and Mo2Oy (y=2, 3, and 4) studied by anion photoelectron spectroscopy and density functional theory calculations. AB - The competitive structural isomers of the Mo(2)O(y) (-)Mo(2)O(y) (y=2, 3, and 4) clusters are investigated using a combination of anion photoelectron (PE) spectroscopy and density functional theory calculations. The PE spectrum and calculations for MoO(3) (-)MoO(3) are also presented to show the level of agreement to be expected between the spectra and calculations. For MoO(3) (-) and MoO(3), the calculations predict symmetric C(3v) structures, an adiabatic electron affinity of 3.34 eV, which is above the observed value 3.17(2) eV. However, there is good agreement between observed and calculated vibrational frequencies and band profiles. The PE spectra of Mo(2)O(2) (-) and Mo(2)O(3) (-) are broad and congested, with partially resolved vibrational structure on the lowest energy bands observed in the spectra. The electron affinities (EA(a)s) of the corresponding clusters are 2.24(2) and 2.33(7) eV, respectively. Based on the calculations, the most stable structure of Mo(2)O(2) (-) is Y shaped, with the two Mo atoms directly bonded. Assignment of the Mo(2)O(3) (-) spectrum is less definitive, but a O-Mo-O-Mo-O structure is more consistent with overall electronic structure observed in the spectrum. The PE spectrum of Mo(2)O(4) (-) shows cleanly resolved vibrational structure and electronic bands, and the EA of the corresponding Mo(2)O(4) is determined to be 2.13(4) eV. The structure most consistent with the observed spectrum has two oxygen bridge bonds between the Mo atoms. PMID- 15836135 TI - Small para-hydrogen clusters doped with carbon monoxide: quantum Monte Carlo simulations and observed infrared spectra. AB - The structures and rotational dynamics of clusters of a single carbon monoxide molecule solvated in para-hydrogen, (paraH(2))(N)-CO, have been simulated for sizes up to N=17 using the reptation Monte Carlo technique. The calculations indicate the presence of two series of R(0) rotational transitions with J=1<--0 for cold clusters, similar to those predicted and observed in the case of He(N) CO. Infrared spectra of these clusters have been observed in the region of the C O stretch ( approximately 2143 cm(-1)) in a pulsed supersonic jet expansion using a tunable diode laser probe. With the help of the calculations, the observed R(0) rotational transitions have been assigned up to N=9 for the b-type series and N=14 for the a-type series. Theory and experiment agree rather well, except that theory tends to overestimate the b-type energies. The (paraH(2))(12)-CO cluster is calculated to be particularly stable and (relatively) rigid, corresponding to completion of the first solvation shell, and it is observed to have the strongest a-type transition. PMID- 15836136 TI - Mutual orientation of two C60 molecules: an ab initio study. AB - The orientational dependence of the interaction between two C(60) molecules is investigated using ab initio calculations. The binding energy, computed within density functional theory in the local density approximation, is substantially smaller than the one derived from the experimental heat of sublimation of fullerite, which calls into question the nature of inter-C(60) bonding. According to our calculations, the experimentally observed orientation with a C(60) presenting a hexagon-hexagon bond to a pentagonal face of the other C(60) is not really favored. Some other configurations are very close in energy and in fact a pentagon facing a pentagon and a hexagon facing a hexagon-hexagon bond are found to be slightly more favorable situations. Our results are compared to previous ones obtained either with previous empirical intermolecular potentials or to existing ab initio studies of crystalline C(60). In addition, the stacking of C(60) in a crystal and in a decahedral (C(60))(7) cluster is discussed. PMID- 15836137 TI - Quantum transition state theory for the full three-dimensional H+H2 reaction. AB - A recently developed quantum transition state theory (QTST) [E. Pollak and J. L. Liao, J. Chem. Phys. 108, 2733 (1998)] for calculating thermal rate constants of chemical reactions is applied to the full three-dimensional hydrogen exchange reaction. Results are compared with other numerical results, for temperatures ranging from T=300 K to T=1500 K. The QTST rate is almost exact at high temperature and is 20% greater than the exact rate at T=300 K, where there is extensive tunneling. PMID- 15836138 TI - The effect of zero-point energy differences on the isotope dependence of the formation of ozone: a classical trajectory study. AB - The effect of zero-point energy differences (DeltaZPE) between the possible fragmentation channels of highly excited O(3) complexes on the isotope dependence of the formation of ozone is investigated by means of classical trajectory calculations and a strong-collision model. DeltaZPE is incorporated in the calculations in a phenomenological way by adjusting the potential energy surface in the product channels so that the correct exothermicities and endothermicities are matched. The model contains two parameters, the frequency of stabilizing collisions omega and an energy dependent parameter Delta(damp), which favors the lower energies in the Maxwell-Boltzmann distribution. The stabilization frequency is used to adjust the pressure dependence of the absolute formation rate while Delta(damp) is utilized to control its isotope dependence. The calculations for several isotope combinations of oxygen atoms show a clear dependence of relative formation rates on DeltaZPE. The results are similar to those of Gao and Marcus [J. Chem. Phys. 116, 137 (2002)] obtained within a statistical model. In particular, like in the statistical approach an ad hoc parameter eta approximately 1.14, which effectively reduces the formation rates of the symmetric ABA ozone molecules, has to be introduced in order to obtain good agreement with the measured relative rates of Janssen et al. [Phys. Chem. Chem. Phys. 3, 4718 (2001)]. The temperature dependence of the recombination rate is also addressed. PMID- 15836139 TI - Fragmentation dynamics of H2S following S 2p photoexcitation. AB - The fragmentation dynamics of core-excited H(2)S has been studied by means of partial anion and cation yield measurements around the S L(2,3)-subshell ionization thresholds. All detectable ionic fragments are reported, and significant differences between partial ion yields are observed. Possible dissociation pathways are discussed by comparison to previous studies of electron spectra. PMID- 15836140 TI - The principles of infrared-x-ray pump-probe spectroscopy. Applications on proton transfer in core-ionized water dimers. AB - In this paper we derive the basic physics underlying infrared-x-ray pump-probe spectroscopy (IR, infrared). Particular features of the spectroscopy are highlighted and discussed, such as dependence on phase of the infrared pulse, duration and delay time of the x-ray pulse, and molecular orientation. Numerical applications are carried out for the water dimer using wave packet techniques. It is shown that core ionization of the donor oxygen of the water dimer results in a drastic change of the potential with the global minimum placed in the proton transfer region. The results of the modeling indicate that IR-x-ray pump-probe spectroscopy can be used to study the dynamics of proton transfer in this core ionized state, and that, contrary to conventional core level photoelectron spectroscopy, x-ray core-ionization driven by an IR field is a proper method to explore the proton transfer in a system like the water dimer. We observe that the trajectory of the nuclear wave packet in the ground state potential well is strongly affected by the absolute phase of the IR pulse. PMID- 15836141 TI - Infrared overtone spectroscopy and unimolecular decay dynamics of peroxynitrous acid. AB - Peroxynitrous acid (HOONO) is generated in a pulsed supersonic expansion through recombination of photolytically generated OH and NO(2) radicals. A rotationally resolved infrared action spectrum of HOONO is obtained in the OH overtone region at 6971.351(4) cm(-1) (origin), providing definitive spectroscopic identification of the trans-perp (tp) conformer of HOONO. Analysis of the rotational band structure yields rotational constants for the near prolate asymmetric top, the ratio of the a-type to c-type components of the transition dipole moment for the hybrid band, and a homogeneous linewidth arising from intramolecular vibrational energy redistribution and/or dissociation. The quantum state distribution of the OH (nu=0,J(OH)) products from dissociation is well characterized by a microcanonical statistical distribution constrained only by the energy available to products, 1304+/-38 cm(-1). This yields a 5667+/-38 cm(-1) [16.2(1) kcal mol( 1)] binding energy for tp-HOONO. An equivalent available energy and corresponding binding energy are obtained from the highest observed OH product state. Complementary high level ab initio calculations are carried out in conjunction with second-order vibrational perturbation theory to predict the spectroscopic observables associated with the OH overtone transition of tp-HOONO including its vibrational frequency, rotational constants, and transition dipole moment. The same approach is used to compute frequencies and intensities of multiple quantum transitions that aid in the assignment of weaker features observed in the OH overtone region, in particular, a combination band of tp-HOONO involving the HOON torsional mode. PMID- 15836142 TI - Photoelectron spectroscopy of fullerene dianions C76(2-), C78(2-), and C84(2-). AB - We report laser photoelectron spectra of the doubly negatively charged fullerenes C(76) (2-), C(78) (2-), and C(84) (2-) at 2.33, 3.49, and 4.66 eV photon energy. From these spectra, second electron affinities and vertical detachment energies, as well as estimates for the repulsive Coulomb barriers are obtained. These results are discussed in the context of electrostatic models. They reveal that fullerenes are similar to conducting spheres, with electronic properties scaling with their size. The experimental spectra are compared with the accessible excited states of the respective singly charged product ions calculated in the framework of time dependent density functional theory. PMID- 15836143 TI - Approaching the basis set limit for transition metal compounds with highly polar bonds: a benchmark coupled-cluster study of the ScF3 and FeF3 molecular structures and spectra. AB - The molecular equilibrium geometries, quadratic and cubic force constants, vibrational frequencies, and infrared intensities of scandium and iron trifluorides are determined ab initio with a sequence of atomic natural orbital basis sets using the CCSD(T) treatment of electron correlation. The largest basis set of spdf ghi quality contains 462 contracted Gaussian functions. Relativistic corrections are applied to compute the equilibrium geometries and vibrational frequencies. The cubic force constants are used to estimate vibrational corrections to the effective r(g) internuclear distances determined in the gas electron diffraction experiments. The computed molecular properties are extrapolated to the complete basis-set limit. The predicted values are compared to the available experimental data; uncertainties and inconsistencies in these data are then discussed. PMID- 15836144 TI - Density functional theory based molecular-dynamics study of aqueous fluoride solvation. AB - We use density functional theory based molecular-dynamics simulations to study the aqueous solvation of the fluoride anion. Our studies are focused on the first solvation shell and have resulted in detailed information on its structural and dynamical properties. The fluoride ion leads to the formation of a rigid solvation shell, qualitatively consistent with simulation and experimental studies, classifying fluoride as a "structure making" particle. However, quantitatively we find the solvation shell to be less structured and more mobile than predicted from empirical force-field simulation. The influence on the intramolecular electronical and structural properties of water is minimal, as observed for other halogens. We propose two distinct mechanisms for the exchange of bulk and first solvation shell water molecules. PMID- 15836145 TI - Diffusion coefficient of ionic solvation shell molecules. AB - It is shown that, for a tightly bound ion-solvation shell complex, the mean square displacement for solvation molecules is characterized by a long lasting transitory. This initial portion is related to the rotational relaxation of the complex and can reach up to several hundred picoseconds for a representative example such as the Mg(2+) ion in water. As the diffusion coefficient is usually fitted using much shorter time spans, unnoticed overestimations are possible. It is argued that, instead of computing the aforementioned diffusion coefficient from the mean square displacement, it should be defined taking as a basic guideline the ratio between the rotational relaxation time of the complex and the lifetime within the first solvation shell. PMID- 15836146 TI - Stress anisotropy induced by periodic boundary conditions. AB - Finite size effects due to periodic boundary conditions are investigated using computer simulations in the canonical ensemble. We study liquids with densities corresponding to typical liquid coexistence densities, and temperatures between the triple and critical points. The components of the pressure tensor are computed in order to analyze the finite size effects arising from the size and geometry of the simulation box. Two different box geometries are considered: cubic and parallelepiped. As expected the pressure tensor is isotropic in cubic boxes, but it becomes anisotropic for small noncubic boxes. We argue this is the origin of the anomalous behavior observed recently in the computation of the surface tension of liquid-vapor interfaces. Otherwise, we find that the bulk pressure is sensitive to the box geometry when small simulation boxes are considered. These observations are general and independent of the model liquid considered. We report results for liquids interacting through short range forces, square well and Lennard-Jones, and also long range Coulombic interactions. The effect that small surface areas have on the surface tension is discussed, and some preliminary results at the liquid vapor-interface for the square well potential are given. PMID- 15836147 TI - Simulations of proton order and disorder in ice Ih. AB - Computer simulations of ice Ih with different proton orientations are presented. Simulations of proton disordered ice are carried out using a Monte Carlo method which samples over proton degree of freedom, allowing for the calculation of the dielectric constant and for the examination of the degree of proton disorder. Simulations are also presented for two proton ordered structures of ice Ih, the ferroelectric Cmc2(1) structure or ice XI and the antiferroelectric Pna2(1) structure. These simulations indicate that a transition to a proton ordered phase occurs at low temperatures (below 80 K). The symmetry of the ordered phase is found to be dependent on the water potential. The stability of the two proton ordered structures is due to a balance of short-ranged interactions which tend to stabilize the Pna2(1) structure and longer-range interactions which stabilize the Cmc2(1) structure. PMID- 15836148 TI - Structural evolution of aqueous NaCl solutions dissolved in supercritical carbon dioxide under isobaric heating by mid and near infrared spectroscopy. AB - The local order in aqueous NaCl solutions diluted in supercritical carbon dioxide at constant pressure as a function of NaCl concentration and temperature has been investigated using near and mid infrared absorption spectroscopy. The near IR results have allowed us to estimate the water concentration in CO(2) rich phase, whereas the state of water aggregation in CO(2) phase was investigated using mid IR spectroscopy. The analysis of the band shape variations of the OD stretching mode of HOD led us to conclude that below 100 degrees C, water molecules dissolved in CO(2) exist only under their monomeric form, whatever the salt concentration is, whereas hydrogen-bonded species, namely, dimers start to appear at higher temperatures. Larger aggregates have a negligible concentration in the range of temperature-pressure investigated. Using near and mid infrared data, we have calculated the concentrations of water species in the CO(2) phase. Upon heating, it was found that the concentration of dimers considerably increases at the expense of the monomers and only dimers are detected in carbon dioxide at highest temperatures. Changing the salt concentration affects significantly the concentration of monomers and decreases strongly the dimers population as the solution becomes progressively saturated in salt. In the saturated solution, at 340 degrees C, the dimer concentration is at least two times smaller than in the binary water-CO(2) mixture. These findings are in qualitative agreement with existing thermodynamics data showing that addition of NaCl to the binary H(2)O CO(2) system shifts the range of partial miscibility of water and CO(2) towards higher pressure and temperature. PMID- 15836149 TI - Microstructure of inhomogeneous polyatomic mixtures from a density functional formalism for atomic mixtures. AB - A free energy density functional theory (DFT) for inhomogeneous polymeric mixtures is developed by treating the polyatomic system as a strongly associating atomic fluid mixture. The theory, derived in terms of segment density, retains the simple form of the DFTs for atomic fluids. Invoking the complete bonding limit of a stoichiometric mixture in the association free energy functional yields a computationally simple and accurate functional for the polyatomic system. Comparisons of theory calculations with molecular simulations are presented for inhomogeneous solutions and blends of linear and branched chains, demonstrating the capability of the theory to accurately capture the entropic and enthalpic effects governing the microstructure. PMID- 15836150 TI - A simple model of entropy relaxation for explaining effective activation energy behavior below the glass transition temperature. AB - Strong changes in relaxation rates observed at the glass transition region are frequently explained in terms of a physical singularity of the molecular motions. We show that the unexpected trends and values for activation energy and preexponential factor of the relaxation time tau, obtained at the glass transition from the analysis of the thermally stimulated current signal, result from the use of the Arrhenius law for treating the experimental data obtained in nonstationary experimental conditions. We then demonstrate that a simple model of structural relaxation based on a time dependent configurational entropy and Adam Gibbs relaxation time is sufficient to explain the experimental behavior, without invoking a kinetic singularity at the glass transition region. The pronounced variation of the effective activation energy appears as a dynamic signature of entropy relaxation that governs the change of relaxation time in nonstationary conditions. A connection is demonstrated between the peak of apparent activation energy measured in nonequilibrium dielectric techniques, with the overshoot of the dynamic specific heat that is obtained in calorimetry techniques. PMID- 15836151 TI - Application of the Wolf damped Coulomb method to simulations of SiC. AB - A multibody interatomic potential is developed for bulk SiC using a modification of the Wolf et al. summation technique [D. Wolf, P. Keblinski, S. R. Phillpot, and J. Eggebrecht, J. Chem. Phys. 110, 8254 (1999)] for the electrostatic interaction. The technique is modified to account for the short-range nonpoint charge effect. The nonelectrostatic interaction is modeled by a simple Morse stretch term. This potential is then applied to beta-SiC to calculate various bulk properties using molecular dynamics simulations. The simulated x-ray diffraction pattern, radial distribution functions, lattice constant, elastic constants, and defect energy agree well with experimental data. PMID- 15836152 TI - Hydrophobic effects on partial molar volume. AB - The hydrophobic effects on partial molar volume (PMV) are investigated as a PMV change in the transfer of a benzenelike nonpolar solute from the nonpolar solvent to water, using an integral equation theory of liquids. The volume change is divided into two effects. One is the "packing" effect in the transfer from the nonpolar solvent to hypothetical "nonpolar water" without hydrogen bonding networks. The other is the "iceberg" effect in the transfer from nonpolar water to water. The results indicate that the packing effect is negative and a half compensated by the positive iceberg effect. The packing effect is explained by the difference in the solvent compressibility. Further investigation shows that the sign and magnitude of the volume change depend on the solute size and the solvent compressibility. The finding gives a significant implication that the exposure of a hydrophobic residue caused by protein denaturation can either increase or decrease the PMV of protein depending on the size of the residue and the fluctuation of its surroundings. PMID- 15836153 TI - New spectroscopic method for aqueous solutions: Raman xi-function dispersion for NaClO4 in water. AB - A new Raman method is exemplified by xi identical with-RT[ partial differential ln(I(omega)I(REF)) partial differentialX(1)](T,P,n(2),n(3) ) for ternary NaClO(4)D(2)OH(2)O, or by xi identical with-RT[ partial differential ln(I(omega)I(REF)) partial differentialX(2)](T,P) for binary NaClO(4)H(2)O solutions. (Fundamental differences exist between xi and the chemical potential mu.) I(omega) is the Raman intensity at omega, I(REF) is the reference intensity, e.g., at the isosbestic frequency, X(2) is the H(2)O and X(1) the small D(2)O mol fraction, and n(2) and n(3) are constant mols of H(2)O and NaClO(4), respectively. Maxima (max) and minima (min) were observed in xi versus omega (cm( 1)); xi(max)-xi(min)=Deltaxi(max). Deltaxi(max)=8050+/-100 calmol H(2)O for the coupled, binary solution OH stretch, and Deltaxi(max)=4200+/-200 calmol H bond for the decoupled, ternary solution OD stretch. The perchlorate ion breaks the H bonds in water. 8050 calmol H(2)O corresponds to the maximum tetrahedral Deltaxi(max) value for two H bonds, i.e., Deltaxi(max)=4025 calmol H bond, in agreement with the HDO Deltaxi(max)=4200+/-200 calmol H bond. [Deltaxi(max) is not the H bond enthalpy (energy).] Minima occur in xi at the peak omega values corresponding to the HDOH(2)O and H(2)O ices, and maxima in xi at 2637+/-5 cm(-1) (OD) and 3575+/-10 cm(-1) (OH) correspond to the peak OD and OH stretching omega values from dense supercritical water. Enthalpy dispersion curves were also determined for saturated, binary, and ternary NaClO(4) solutions and for D(2)O in H(2)O. The xi-function method is shown to be applicable to infrared absorbance spectra. PMID- 15836154 TI - Incorporating variable dielectric environments into the generalized Born model. AB - A generalized Born (GB) model is proposed that approximates the electrostatic part of macromolecular solvation free energy over the entire range of the solvent and solute dielectric constants. The model contains no fitting parameters, and is derived by matching a general form of the GB Green function with the exact Green's function of the Poisson equation for a random charge distribution inside a perfect sphere. The sphere is assumed to be filled uniformly with dielectric medium epsilon(in), and is surrounded by infinite solvent of constant dielectric epsilon(out). This model is as computationally efficient as the conventional GB model based on the widely used functional form due to Still et al. [J. Am. Chem. Soc. 112, 6127 (1990)], but captures the essential physics of the dielectric response for all values of epsilon(in) and epsilon(out). This model is tested against the exact solution on a perfect sphere, and against the numerical Poisson Boltzmann (PB) treatment on a set of macromolecules representing various structural classes. It shows reasonable agreement with both the exact and the numerical solutions of the PB equation (where available) considered as reference, and is more accurate than the conventional GB model over the entire range of dielectric values. PMID- 15836155 TI - Homogeneous nucleation of n-propanol, n-butanol, and n-pentanol in a supersonic nozzle. AB - We have measured the nucleation conditions of n-propanol, n-butanol, and n pentanol in a supersonic Laval nozzle, and estimated that the maximum nucleation rate J is 5 x 10(16) cm(-3) s(-1) with an uncertainty factor of 2. Plotting the vapor pressures p(J(max) ) and temperatures T(J(max) ) corresponding to the maximum nucleation rate as ln(p) versus 1T, produces a series of well separated straight lines. When these values are scaled by their respective critical parameters, p(c) and T(c), the data lie close to a single straight line. Comparing the experimental data to the predictions of classical nucleation theory reveals much higher experimental rates, and the deviation increases with increasing alcohol chain length and decreasing temperature. A scaling analysis in terms of Hale's scaled nucleation model [Phys. Rev. A 33, 4156 (1986); Metall. Trans. A 23, 1863 (1992)], clearly shows that our data are consistent with experimental nucleation rates measured using other devices that have characteristic rates many orders of magnitude lower. PMID- 15836156 TI - Absolute vibrational and electronic cross sections for low-energy electron (2-12 eV) scattering from condensed pyrimidine. AB - Low-energy vibrational and electronic electron-energy-loss (EEL) spectra of pyrimidine condensed on a thin film of solid argon held at 18 K are reported for the incident-energy range of 2-12 eV. Sensitivity to symmetry and spin forbidden transitions as well as correlations to the triplet states of benzene make it possible to ascribe the main features, below 7 eV in the electronic part of the EEL spectrum, to triplet transitions. The lowest EEL feature with an energy onset at 3.5 eV is attributed to a transition to the (3)B(1)(n-->pi(*)) valence electronic state and the next triplet n-->pi(*) transition to a (3)A(2) state located around 4.5 eV. The remaining EEL features at 4.3, 5.2, 5.8, and 6.5 eV are all assigned to pi-->pi(*) transitions to states of symmetry (3)B(2), (3)A(1), (3)B(2), and (3)B(2)+(3)A(1), respectively. The most intense maximum at 7.6 eV is found to correspond to both (1)B(2) and (1)A(1) transitions, as in the vacuum ultraviolet spectra. Absolute inelastic cross sections per scatterer are derived from a single collision treatment described herein. Their values are found to lie within the 10(-17) cm(2) range for both the electronic and the vibrational excitations. Features in the energy dependence of the cross sections are discussed, whenever possible, by comparison with data and mechanisms found in the gas phase. A maximum over the 4-5 eV range is attributed to a B (2)B(1) shape resonance and another one observed in the 6-7 eV range is ascribed to either or both sigma(*) shape resonances of (2)A(1) and (2)B(2) symmetries. PMID- 15836157 TI - Restricted orientational motion of nitroxides in molecular glasses: direct estimation of the motional time scale basing on the comparative study of primary and stimulated electron spin echo decays. AB - A comparative study of anisotropic relaxation in two-pulse primary and three pulse stimulated electron spin echo decays provides a direct way to distinguish fast (correlation time tau(c)<10(-6) s) and slow (tau(c)>10(-6) s) motions. Anisotropic relaxation is detected as a difference of the decay rates for different resonance field positions in anisotropic electron paramagnetic resonance spectra. For fast motion anisotropic relaxation influences the primary echo decay and does not influence the stimulated echo decay. For slow motion it is seen in both two-pulse echo and three-pulse stimulated echo decays. For nitroxide spin probes dissolved in glassy glycerol only fast motion was found below 200 K. Increase of temperature above 200 K results in the appearance of slow motion. Its amplitude increases rapidly with temperature increase. While in glycerol glass slow motion appears above glass transition temperature T(g), in ethanol glass it is observable below T(g). The scenario of motional dynamics in glasses is proposed which involves the broadening of the correlation time distribution with increasing temperature. PMID- 15836158 TI - Two-dimensional connective nanostructures of electrodeposited Zn on Au (111) induced by spinodal decomposition. AB - Phase formation of surface alloying by spinodal decomposition has been studied at an electrified interface. For this aim Zn was electrodeposited on Au(111) from the ionic liquid AlCl(3)-MBIC (58:42) containing 1 mM Zn(II) at different potentials in the underpotential range corresponding to submonolayer up to monolayer coverage. Structure evolution was observed by in situ electrochemical scanning tunneling microscopy (STM) at different times after starting the deposition via potential jumps and at temperatures of 298 and 323 K. Spinodal or labyrinth two-dimensional structures predominate at middle coverage, both in deposition and in dissolution experiments. They are characterized by a length scale of typically 5 nm which has been determined from the power spectral density of STM images. Structure formation and surface alloying are governed by slow kinetics with a rate constant k with activation energy of 120 meV and preexponential factor of 0.17 s(-1). The evolution of the structural features is described by a continuum model and is found to be in good agreement with the STM observations. From the experimental and model calculation results we conclude that the two-dimensional phase formation in the Zn on Au(111) system is dominated by surface alloying. The phase separation of a Zn-rich and a Zn-Au alloy phase is governed by two-dimensional spinodal decomposition. PMID- 15836159 TI - Water adsorption in disordered mesoporous silica (Vycor) at 300 K and 650 K: a Grand Canonical Monte Carlo simulation study of hysteresis. AB - This numerical simulation paper focuses on the adsorption/desorption of water in disordered mesoporous silica glasses (Vycor-like). The numerical adsorbent was previously obtained by off lattice method, and was shown to reproduce quite well the micro- and mesotextural properties of real Vycor, as well as morphological (pore size distribution) and topological (pore interconnections) disorder. The water-water interactions are described by the SPC model while water-silica interactions are calculated in the framework of the PN-TrAZ model. The water adsorption/desorption isotherms and the configurational energies are calculated by the Grand Canonical Monte Carlo simulation method. The low pressure results compare well with experiments, showing the good transferability of the intermolecular potential. It is shown that if the hysteresis loop observed in the adsorption/desorption isotherm is considered as a true phase transition (which is actually still an open question in the case of disordered porous materials), then it is possible to calculate the grand potential by applying the thermodynamic integration scheme. The grand potential is shown to be multivalued for low (subcritical) temperature, and continuous for high (supercritical) temperature. A coexistence point is found within the hysteresis loop, actually close to the vertical desorption line. Below the equilibrium chemical potential, the gaslike branch is stable whereas the liquidlike branch is metastable. The situation is reversed above the coexistence point. PMID- 15836160 TI - Detailed characterization of (3 x 3) iodine adlayer on Pt(111) by unequal-sphere packing model. AB - A simple unequal-sphere packing model is applied to study the iodine (3x3) adlayer on the Pt(111) surface. By using a newly introduced parameter, defined as the average adsorbate height, three characteristic adlattices, (3x3)-sym, (3x3) asym, and (3x3)-lin, have been selected and characterized in great detail, including the exact adatom registry. The (3x3)-sym iodine adlattice, observed in many experimental studies, appears to be, on average, the closest one to the substrate surface. A special contour plot of average adsorbate height vs X and Y positions of the (3x3) iodine unit cell indicates the existence of two local minima, which are related to preferential formation of (3x3)-sym and (3x3)-asym iodine adlattices. Our model gives good agreement with experimental findings, and explains the mechanism of preferential appearance of (3x3)-sym and (3x3)-asym structures. PMID- 15836161 TI - Role of molecular orbitals of the benzene in electronic nanodevices. AB - In an effort to examine the intricacies of electronic nanodevices, we present an atomistic description of the electronic transport properties of an isolated benzene molecule. We have carried out ab initio calculations to understand the modulation of the molecular orbitals (MOs) and their energy spectra under the external electric field, and conducting behavior of the benzene molecule. Our study shows that with an increase in the applied electric field, the energy of the third lowest unoccupied molecular orbital (LUMO) of benzene decreases, while the first and second LUMO energies are not affected. Above a certain threshold of the external electric field, the third LUMO is lowered below the original LUMO and becomes the real LUMO. Since the transport through a molecule is to a large extent mediated by the molecular orbitals, the change in MOs can lead to a dramatic increase in the current passing through the benzene molecule. Thus, in the course of this study, we show that the modulation of the molecular orbitals in the presence of a tuning parameter(s) such as the external electric field can play important roles in the operation of molecular devices. We believe that this understanding would be helpful in the design of electronic nanodevices. PMID- 15836162 TI - Flow curves of dense colloidal dispersions: schematic model analysis of the shear dependent viscosity near the colloidal glass transition. AB - A recently proposed schematic model for the nonlinear rheology of dense colloidal dispersions is compared to flow curves measured in suspensions that consist of thermosensitive particles. The volume fraction of this purely repulsive model system can be adjusted by changing temperature. Hence, high volume fractions (phi2 (3)P(2) transition. It is argued that relevant comparisons between nonrelativistic and relativistic calculations must be performed at the level of integrated absorption cross sections. PMID- 15836201 TI - Leading-order relativistic effects on nuclear magnetic resonance shielding tensors. AB - We present perturbational ab initio calculations of the nuclear-spin-dependent relativistic corrections to the nuclear magnetic resonance shielding tensors that constitute, together with the other relativistic terms reported by us earlier, the full leading-order perturbational set of results for the one-electron relativistic contributions to this observable, based on the (Breit-)Pauli Hamiltonian. These contributions are considered for the H(2)X (X = O,S,Se,Te,Po) and HX (X = F,Cl,Br,I,At) molecules, as well as the noble gas (Ne, Ar, Kr, Xe, Rn) atoms. The corrections are evaluated using the relativistic and magnetic operators as perturbations on an equal footing, calculated using analytical linear and quadratic response theory applied on top of a nonrelativistic reference state provided by self-consistent field calculations. The (1)H and heavy-atom nuclear magnetic shielding tensors are compared with four component, nearly basis-set-limit Dirac-Hartree-Fock calculations that include positronic excitations, as well as available literature data. Besides the easy interpretability of the different contributions in terms of familiar nonrelativistic concepts, the accuracy of the present perturbational scheme is striking for the isotropic part of the shielding tensor, for systems including elements up to Xe. PMID- 15836202 TI - Free energy profile along a discretized reaction path via the hyperplane constraint force and torque. AB - By employing mechanical work analogies, we derive a convenient computational approach for evaluation of the free energy profile (FEP) along some discretized path defined as a sequence of hyperplanes. A hyperplane is fully specified by any of its point and a tangent vector. The FEP is obtained as an integral of two components. The translational component of the free energy is computed by integrating the hyperplane constraint force. The rotational component is evaluated via the hyperplane torque. Both ingredients--the constraint force and the hyperplane torque-are evaluated on each hyperplane independently. The integration procedure utilizes a set of reference points defining a point of rotation on each hyperplane, and these points can be chosen before or after the sampling takes place. A shift in the reference points redistributes the FEP contributions between the translational and rotational components. For systems where the FEP is dominated by the potential energy differences, reference points residing on the minimum energy path present a natural choice. We demonstrate the validity of our approach on two examples, a simple two-dimensional (2D) potential, and a seven-atom Lennard-Jones cluster. In each case, we compare the numerical FEP with the harmonic approximation estimates. Our results for the 2D potential are also verified by the data available in the literature. In both cases, the rotational component of the FEP represents a sizable contribution to the total FEP, so ignoring it would yield clearly incorrect results. PMID- 15836203 TI - On dynamical tunneling and classical resonances. AB - This work establishes a firm relationship between classical nonlinear resonances and the phenomenon of dynamical tunneling. It is shown that the classical phase space with its hierarchy of resonance islands completely characterizes dynamical tunneling and explicit forms of the dynamical barriers can be obtained only by identifying the key resonances. Relationship between the phase space viewpoint and the quantum mechanical superexchange approach is discussed in near-integrable and mixed regular-chaotic situations. For near-integrable systems with sufficient anharmonicity the effect of multiple resonances, i.e., resonance-assisted tunneling, can be incorporated approximately. It is also argued that the presumed relation of avoided crossings to nonlinear resonances does not have to be invoked in order to understand dynamical tunneling. For molecules with low density of states the resonance-assisted mechanism is expected to be dominant. PMID- 15836204 TI - Validation of the density-functional based tight-binding approximation method for the calculation of reaction energies and other data. AB - We investigated the performance of the approximative density functional method DFTB versus BLYP and G2 with respect to zero-point corrected reaction energies, vibrational frequencies, and geometry parameters for a set of 28 reactions and 22 representative molecules containing C, H, N, and O (DFTB--density-functional based tight-binding approximation). The DFTB reaction energies show a mean absolute deviation versus the G2 reference of 4.3 kcalmol only. The corresponding value for the vibrational frequencies amounts to 75 cm(-1) versus BLYP/cc-pVTZ. With very few exceptions bond lengths and angles are in excellent agreement with the results of higher-level methods. PMID- 15836205 TI - Dissipating the Langevin equation in the presence of an external stochastic potential. AB - In the Langevin formalism, the delicate balance maintained between the fluctuations in the system and their corresponding dissipation may be upset by the presence of a secondary, space-dependent stochastic force, particularly in the low-friction regime. In prior work, the latter was dissipated self consistently through an additional uniform (mean-field) friction [T. Shepherd and R. Hernandez, J. Chem. Phys. 115, 2430 (2001).] An alternative approach to ensure that equipartition is satisfied relies on the use of a space-dependent friction while ignoring nonlocal correlations. The approach is evaluated with respect to its ability to maintain constant temperature for two simple one-dimensional, stochastic potentials of mean force wherein the friction can be evaluated explicitly when there is no memory in the barriers. The use of a space-dependent friction is capable of providing qualitatively similar results to those obtained previously, but in extreme cases deviations from equipartition may be observed due to the neglect of the memory effects present in the stochastic potentials. PMID- 15836206 TI - From molecular dynamics to hydrodynamics: a novel Galilean invariant thermostat. AB - This paper proposes a novel thermostat applicable to any particle-based dynamic simulation. Each pair of particles is thermostated either (with probability P) with a pairwise Lowe-Andersen thermostat [C. P. Lowe, Europhys. Lett. 47, 145 (1999)] or (with probability 1-P) with a thermostat that is introduced here, which is based on a pairwise interaction similar to the Nose-Hoover thermostat. When the pairwise Nose-Hoover thermostat dominates (low P), the liquid has a high diffusion coefficient and low viscosity, but when the Lowe-Andersen thermostat dominates, the diffusion coefficient is low and viscosity is high. This novel Nose-Hoover-Lowe-Andersen thermostat is Galilean invariant and preserves both total linear and angular momentum of the system, due to the fact that the thermostatic forces between each pair of the particles are pairwise additive and central. We show by simulation that this thermostat also preserves hydrodynamics. For the (noninteracting) ideal gas at P = 0, the diffusion coefficient diverges and viscosity is zero, while for P > 0 it has a finite value. By adjusting probability P, the Schmidt number can be varied by orders of magnitude. The temperature deviation from the required value is at least an order of magnitude smaller than in dissipative particle dynamics (DPD), while the equilibrium properties of the system are very well reproduced. The thermostat is easy to implement and offers a computational efficiency better than (DPD), with better temperature control and greater flexibility in terms of adjusting the diffusion coefficient and viscosity of the simulated system. Applications of this thermostat include all standard molecular dynamic simulations of dense liquids and solids with any type of force field, as well as hydrodynamic simulation of multiphase systems with largely different bulk viscosities, including surface viscosity, and of dilute gases and plasmas. PMID- 15836207 TI - Combining smart darting with parallel tempering using Eckart space: application to Lennard-Jones clusters. AB - The smart-darting algorithm is a Monte Carlo based simulation method used to overcome quasiergodicity problems associated with disconnected regions of configurations space separated by high energy barriers. As originally implemented, the smart-darting method works well for clusters at low temperatures with the angular momentum restricted to zero and where there are no transitions to permutational isomers. If the rotational motion of the clusters is unrestricted or if permutational isomerization becomes important, the acceptance probability of darting moves in the original implementation of the method becomes vanishingly small. In this work the smart-darting algorithm is combined with the parallel tempering method in a manner where both rotational motion and permutational isomerization events are important. To enable the combination of parallel tempering with smart darting so that the smart-darting moves have a reasonable acceptance probability, the original algorithm is modified by using a restricted space for the smart-darting moves. The restricted space uses a body fixed coordinate system first introduced by Eckart, and moves in this Eckart space are coupled with local moves in the full 3N-dimensional space. The modified smart-darting method is applied to the calculation of the heat capacity of a seven-atom Lennard-Jones cluster. The smart-darting moves yield significant improvement in the statistical fluctuations of the calculated heat capacity in the region of temperatures where the system isomerizes. When the modified smart darting algorithm is combined with parallel tempering, the statistical fluctuations of the heat capacity of a seven-atom Lennard-Jones cluster using the combined method are smaller than parallel tempering when used alone. PMID- 15836208 TI - Matching-pursuit/split-operator Fourier-transform simulations of nonadiabatic quantum dynamics. AB - A rigorous and practical approach for simulations of nonadiabatic quantum dynamics is introduced. The algorithm involves a natural extension of the matching-pursuitsplit-operator Fourier-transform (MPSOFT) method [Y. Wu and V. S. Batista, J. Chem. Phys. 121, 1676 (2004)] recently developed for simulations of adiabatic quantum dynamics in multidimensional systems. The MPSOFT propagation scheme, extended to nonadiabatic dynamics, recursively applies the time-evolution operator as defined by the standard perturbation expansion to first-, or second order, accuracy. The expansion is implemented in dynamically adaptive coherent state representations, generated by an approach that combines the matching pursuit algorithm with a gradient-based optimization method. The accuracy and efficiency of the resulting propagation method are demonstrated as applied to the canonical model systems introduced by Tully for testing simulations of dual curve crossing nonadiabatic dynamics. PMID- 15836209 TI - Experimental and theoretical investigation of the reaction NH(X3Sigma-) + H(2S)- >N(4S) + H2(X1)Sigmag +). AB - The rate coefficient of the reaction NH(X (3)Sigma(-)) + H((2)S)-->(k(1a) )N((4)S) + H(2)(X (1)Sigma(g) (+)) is determined in a quasistatic laser-flash photolysis, laser-induced fluorescence system at low pressures (2 mbar< or =p< or =10 mbar). The NH(X) radicals are produced via the quenching of NH(a(1)Delta) (obtained by photolyzing HN(3)) with Xe whereas the H atoms are generated in a H(2)He microwave discharge. The NH(X) concentration profile is measured under pseudo-first-order condition, i.e., in the presence of a large excess of H atoms. The room temperature rate coefficient is determined to be k(1a) = (1.9 +/- 0.5) x 10(12) cm(3) mol(-1) s(-1). It is found to be independent of the pressure in the range considered in the present experiment. A global potential energy surface for the (4)A(") state is calculated with the internally contracted multireference configuration interaction method and the augmented correlation consistent polarized valence quadruple zeta atomic basis. The title reaction is investigated by classical trajectory calculations on this surface. The theoretical room temperature rate coefficient is k(1a) = 0.92 x 10(12)cm(3) mol(-1) s(-1). Using the thermodynamical data for the atoms and molecules involved, the rate coefficient for the reverse reaction, k(-1a), is also calculated. At high temperatures it agrees well with the measured k(-1a). PMID- 15836210 TI - Accurate potential energy curves for HeO-, NeO-, and ArO-: spectroscopy and transport coefficients. AB - We calculate accurate potential energy curves for HeO(-), NeO(-), and ArO(-), including the full counterpoise correction and allowing for spin-orbit effects. Comparison with previous curves is presented, where these are available. The three curves, (2)Sigma(12) (+), (2)Pi(12), and (2)Pi(32), are used to derive spectroscopic constants and to calculate the transport coefficients for O(-) moving in a bath of the respective rare gas. Conclusions are made based on a comparison with the available data. PMID- 15836211 TI - Novel structures and energy spectra of hydroxylated (SiO2)8-based clusters: searching for the magic (SiO2)8O2H3- cluster. AB - The prominent (SiO(2))(8)O(2)H(3) (-) mass peak resulting from the laser ablation of hydroxylated silica, attributed to magic cluster formation, is investigated employing global optimization with a dedicated interatomic potential and density functional calculations. The low-energy spectra of cluster isomers are calculated for the closed shell clusters: (SiO(2))(8)OH(-) and (SiO(2))(8)O(2)H(3) (-) giving the likely global minima in each case. Based upon our calculated cluster structures and energetics, and further on the known experimental details, it is proposed that the abundant formation of (SiO(2))(8)O(2)H(3) (-) clusters is largely dependent on the high stability of the (SiO(2))(8)OH(-) ground state cluster. Both the (SiO(2))(8)O(2)H(3) (-) and (SiO(2))(8)OH(-) ground state clusters are found to exhibit cagelike structures with the latter containing a particularly unusual tetrahedrally four-coordinated oxygen center not observed before in either bulk silica or silica clusters. The bare ground state (SiO(2))(8)O(2-) cluster ion core is also found to have four tetrahedrally symmetric Si==O terminations making it a possible candidate, when combined with suitable cations, for extended cluster-based structures/materials. PMID- 15836212 TI - Ab initio n-electron valence state perturbation theory study of the adiabatic transitions in carbonyl molecules: formaldehyde, acetaldehyde, and acetone. AB - The application of the recently developed second-order n-electron valence state perturbation theory (NEVPT2) to small carbonyl molecules (formaldehyde, acetaldehyde, and acetone) is presented. The adiabatic transition energies are computed for the singlet and triplet n-->pi(*), pi-->pi(*), and sigma-->pi(*) states performing a full geometry optimization of the relevant states at the single state CASSCF level and taking into account the zero point energy correction in the harmonic approximation. The agreement with the known experimental values and with previously published high level calculations confirms that NEVPT2 is an efficient tool to be used for the interpretation of molecular electronic spectra. Moreover, different insight into the nature of the excited states has been obtained. Some of the transitions presented here have never been theoretically computed previously [(3)(pi-->pi(*)) and (3)(sigma- >pi(*)) adiabatic transitions in acetaldehyde and acetone] or have been studied only using moderate level (single reference based) ab initio methods (all adiabatic transitions in acetaldehyde). In the present work a consistent disagreement between NEVPT2 and experiment has been found for the (3)(pi-->pi(*)) adiabatic transition in all molecules: this result is attributed to the low intensity of the transition to the first vibrational levels of the excited state. The n-->pi(*) singlet and triplet vertical transition energies are also reported for all the molecules. PMID- 15836213 TI - First principles study of the carbon-(silicon-) doped La13 clusters. AB - The structural stability and physical properties have been studied for carbon (silicon-) doped La(13) clusters using DMOL method based on density-functional theory. Doped La(13) clusters prefer to be icosahedron. Substitutional doping with a carbon or silicon impurity makes some clusters closed electronic shell, especially in icosahedral isomers. Substitutional doping of icosahedral La(13) clusters is found to be favorable at surface sites of clusters, especially for Si doped La(13) cluster, which is very likely to be formed during the doping process. In addition, the structural distortions due to the doping are discussed. PMID- 15836214 TI - Molecular alignment of ammonia studied by electron-ion-ion coincidence spectroscopy. AB - Electron-ion-ion coincidence measurements carried out at discrete resonances near the N 1s threshold in ammonia are reported. The measured coincidence spectra show clear alignment of the molecule upon resonant core-electron excitation. The coincidence data are analyzed to extract information about the molecule in the excited state by simulating the alignment and the dissociation processes. Dynamic changes in molecular geometry are found as the photon energy is scanned through the N 1s-->4a(1) resonance, whereas for the N 1s-->2e state the geometry and kinetic energy released upon dissociation remain unchanged. The alignment of the core-excited molecules is found to be preserved even in two-step dissociation processes. PMID- 15836215 TI - Pulsed laser photolysis and quantum chemical-statistical rate study of the reaction of the ethynyl radical with water vapor. AB - The rate coefficient of the gas-phase reaction C(2)H + H(2)O-->products has been experimentally determined over the temperature range 500-825 K using a pulsed laser photolysis-chemiluminescence (PLP-CL) technique. Ethynyl radicals (C(2)H) were generated by pulsed 193 nm photolysis of C(2)H(2) in the presence of H(2)O vapor and buffer gas N(2) at 15 Torr. The relative concentration of C(2)H radicals was monitored as a function of time using a CH* chemiluminescence method. The rate constant determinations for C(2)H + H(2)O were k(1)(550 K) = (2.3 +/- 1.3) x 10(-13) cm(3) s(-1), k(1)(770 K) =(7.2 +/- 1.4) x 10(-13) cm(3) s(-1), and k(1)(825 K) = (7.7 +/- 1.5) x 10(-13) cm(3) s(-1). The error in the only other measurement of this rate constant is also discussed. We have also characterized the reaction theoretically using quantum chemical computations. The relevant portion of the potential energy surface of C(2)H(3)O in its doublet electronic ground state has been investigated using density functional theory B3LYP6-311 + + G(3df,2p) and molecular orbital computations at the unrestricted coupled-cluster level of theory that incorporates all single and double excitations plus perturbative corrections for the triple excitations, along with the 6-311 + + G(3df,2p) basis set [(U)CCSD(T)6-311 + + G(3df,2p)] and using UCCSD(T)6-31G(d,p) optimized geometries. Five isomers, six dissociation products, and sixteen transition structures were characterized. The results confirm that the hydrogen abstraction producing C(2)H(2)+OH is the most facile reaction channel. For this channel, refined computations using (U)CCSD(T)6-311 + + G(3df,2p)(U)CCSD(T)6-311 + + G(d,p) and complete-active-space second-order perturbation theory/complete-active-space self-consistent-field theory (CASPT2/CASSCF) [B. O. Roos, Adv. Chem. Phys. 69, 399 (1987)] using the contracted atomic natural orbitals basis set (ANO-L) [J. Almlof and P. R. Taylor, J. Chem. Phys.86, 4070 (1987)] were performed, yielding zero-point energy corrected potential energy barriers of 17 kJ mol(-1) and 15 kJ mol(-1), respectively. Transition-state theory rate constant calculations, based on the UCCSD(T) and CASPT2/CASSCF computations that also include H-atom tunneling and a hindered internal rotation, are in perfect agreement with the experimental values. Considering both our experimental and theoretical determinations, the rate constant can best be expressed, in modified Arrhenius form as k(1)(T) = (2.2 +/- 0.1) x 10(-21)T(3.05) exp[-(376 +/- 100)T] cm(3) s(-1) for the range 300-2000 K. Thus, at temperatures above 1500 K, reaction of C(2)H with H(2)O is predicted to be one of the dominant C(2)H reactions in hydrocarbon combustion. PMID- 15836216 TI - Ozonolysis of alpha-pinene and beta-pinene: kinetics and mechanism. AB - A combined quantum-chemical and RRKM/ME (ME--master equation) approach is employed to investigate the structures, energetics, and kinetics of intermediate and stable species, and the yields of stabilized carbonyl oxides and OH radicals from the alpha-pinene and beta-pinene ozonolysis reactions. The cycloaddition of O(3) is highly exothermic, with the reaction energies of 55.1 and 51.1 kcal mol( 1) for alpha- and beta-pinenes, respectively. Cleavage of primary ozonides yields carbonyl oxides with the barrier height of 12.2-17.5 kcal mol(-1). For the prompt reactions of carbonyl oxides from alpha- and beta-pinene ozonolysis, H migration to hydroperoxides represents the dominant pathway over ring closure to dioxiranes. The kinetic calculations indicate a significant portion of stabilization for alpha- and beta-carbonyl oxides. The yields of stabilized carbonyl oxides are estimated to be 0.34 for alpha-pinene and 0.22 for beta pinene. The applicability of theoretical methods for investigation of oxidation reactions of large hydrocarbon molecules is demonstrated. PMID- 15836217 TI - Infrared water vapor continuum absorption at atmospheric temperatures. AB - We have used a continuous-wave carbon dioxide laser in a single-mode realization of cavity ring-down spectroscopy to measure absorption coefficients of water vapor at 944 cm(-1) for several temperatures in the range 270-315 K. The conventional description of water vapor infrared absorption is applied, in which the absorption is modeled in two parts consisting of local line absorption and the remaining residual absorption, which has become known as the water vapor continuum. This water vapor continuum consists of distinct water-water, water nitrogen, and water-oxygen continua. The water-water continuum absorption coefficient is found to have a magnitude of C(s)(296 K) = (1.82+/-0.02) x 10(-22) cm(2) molecule(-1) atm(-1), and the water-nitrogen coefficient has a magnitude of C(n)(296 K) = (7.3 +/- 0.4) x 10(-25) cm(2) molecule(-1) atm(-1). The temperature dependences of both the water-water and the water-nitrogen continua are shown to be well represented by a model describing the expected behavior of weakly bound binary complexes. Using this model, our data yield dissociation energies of D(e) = (-15.9 +/- 0.3) kJ/mole for the water dimer and D(e) = (-3.2 +/- 1.7) kJ/mole for the water-nitrogen complex. These values are in excellent agreement with recent theoretical predictions of D(e) = -15.7 kJ/mole (water dimer) and D(e) = 2.9 kJ/mole (water-nitrogen complex), as well as the experimentally determined value of D(e) = (-15.3 +/- 2.1) kJ/mole for the water dimer obtained by investigators employing a thermal conductivity technique. Although there is reasonably good agreement with the magnitude of the continuum absorption coefficients, the agreement on temperature dependence is less satisfactory. While our results are suggestive of the role played by water dimers and water complexes in producing the infrared continuum, the uncertain spectroscopy of the water dimer in this spectral region prevents us from making a firm conclusion. In the meantime, empirical models of water vapor continuum absorption, essential for atmospheric radiative transfer calculations, should be refined to give better agreement with our low-uncertainty continuum absorption data. PMID- 15836218 TI - Geometric, electronic, and bonding properties of AuNM (N = 1-7, M = Ni, Pd, Pt) clusters. AB - Employing first-principles methods, based on density functional theory, we report the ground state geometric and electronic structures of gold clusters doped with platinum group atoms, Au(N)M (N = 1-7, M = Ni, Pd, Pt). The stability and electronic properties of Ni-doped gold clusters are similar to that of pure gold clusters with an enhancement of bond strength. Due to the strong d-d or s-d interplay between impurities and gold atoms originating in the relativistic effects and unique properties of dopant delocalized s-electrons in Pd- and Pt doped gold clusters, the dopant atoms markedly change the geometric and electronic properties of gold clusters, and stronger bond energies are found in Pt-doped clusters. The Mulliken populations analysis of impurities and detailed decompositions of bond energies as well as a variety of density of states of the most stable dopant gold clusters are given to understand the different effects of individual dopant atom on bonding and electronic properties of dopant gold clusters. From the electronic properties of dopant gold clusters, the different chemical reactivity toward O(2), CO, or NO molecule is predicted in transition metal-doped gold clusters compared to pure gold clusters. PMID- 15836219 TI - Theoretical characterization of dihydrogen adducts with alkaline cations. AB - As part of an extended fundamental study on the interaction of molecular hydrogen with the surfaces of potential storage materials, this work describes the interaction of dihydrogen with bare alkaline cations (Li(+),Na(+),K(+),Rb(+)) by means of ab initio and density functional calculations. The effects of method and basis set are evaluated, and an estimate of the electrostatic contributions to the binding energy is evaluated. All methods predict a t-shaped coordination geometry, in accord with the quadrupolar nature of the molecule. The binding energies at the MP2/aug-cc-pVQZ (MP2-Moller-Plesset method truncated at second order) level ranges between 24 kJ/mol for Li(+) and 5 kJ/mol for Rb(+). Basis set size has minor effect on the binding energy of the complexes, although the poorer sets have severe limitations in the description of the electrostatics of the isolated molecule. The electrostatic contribution to the binding energy calculated from the electrostatic properties of the isolated molecule are in fact strongly basis set dependent. Their careful analysis shows that the predominant terms of the interaction are those due to the quadrupole and dipole polarizability terms. The vibrational spectra of the adducts are evaluated and compared with a large set of experimental and theoretical results from the literature. This review highlights a linear correlation between the frequency shift and the binding energy, which is valuable in the understanding of adsorption phenomena by means of spectroscopic methods. PMID- 15836220 TI - Potential-energy surface, dynamics of van der Waals motions, and vibronic transitions in p-difluorobenzene-argon complex. AB - The dynamics of van der Waals vibrational motions and vibronic spectrum of the complex of argon with p-difluorobenzene (ArDFB) are investigated using the ab initio method. The electronic ground-state potential-energy surface of the complex is calculated at the second-order Moller-Plesset level of theory using a well-balanced basis set aug-cc-pVDZ and its reduced version without tight polarization functions. The dissociation energy of 351 cm(-1) and the binding energy of 402 cm(-1) determined at the Ar distance of 3.521 Angstroms from the DFB ring well agree with the experimental data available. The character of calculated vibrational levels is analyzed and the effect of a strong coupling between the stretching and bending motions is investigated. A new class of hybrid states created by this coupling is found. To investigate the vibronic S(1)-S(0) spectrum, the surfaces of the electronic transition dipole moment are calculated using the ab initio method. From these surfaces, the vibronic transition intensities are determined and employed to assign the Franck-Condon- and Herzberg Teller-induced transitions. PMID- 15836221 TI - Quasiclassical trajectory study of formaldehyde unimolecular dissociation: H(2)CO ->H2 + CO, H + HCO. AB - We report quasiclassical trajectory calculations of the dynamics of the two reaction channels of formaldehyde dissociation on a global ab initio potential energy surface: the molecular channel H(2)CO-->H(2) + CO and the radical H(2)CO- >H + HCO. For the molecular channel, it is confirmed that above the threshold of the radical channel a second, intramolecular hydrogen abstraction pathway is opened to produce CO with low rotation and vibrationally hot H(2). The low-j(CO) and high-nu(H(2) ) products from the second pathway increase with the total energy. The competition between the molecular and radical pathways is also studied. It shows that the branching ratio of the molecular products decreases with increasing energy, while the branching ratio of the radical products increases. The results agree well with very recent velocity-map imaging experiments of Suits and co-workers and solves a mystery first posed by Moore and co-workers. For the radical channel, we present the translational energy distributions and HCO rotation distributions at various energies. There is mixed agreement with the experiments of Wittig and co-workers, and this provides an indirect confirmation of their speculation that the triplet surface plays a role in the formation of the radical products. PMID- 15836222 TI - High resolution emission Fourier transform infrared spectra of the 4p-5s and 5p 6s bands of ArH. AB - In the 2500-8500 cm(-1) region several strong emission bands of (40)ArH were observed by Fourier transform spectroscopy through a dc glow discharge in a mixture of argon and hydrogen. Rotational-electronic transitions of the two previously unstudied 4p-5s and 5p-6s,v = 0-0, bands of (40)ArH were measured and assigned in the 6060 and 3770 cm(-1) regions, respectively. A simultaneous fit of the emission transitions of the 4p-5s and 5p-6s bands and an extended set of transitions of the 6s-4p band observed by Dabrowski, Tokaryk, and Watson [J. Mol. Spectrosc. 189, 95 (1998)] and remeasured in the present work yielded consistent values of the spectroscopic parameters of the electronic states under investigation. In the branch of the 4p-5s band with transitions of type (Q)Q(f(3)e) we observed a narrowing in the linewidths with increasing rotational quantum number N. The rotational dependence of the linewidth is caused by predissociation of the 5s state by the repulsive ground 4s state through homogeneous coupling and changes in overlap integrals of the vibrational wave functions with the rotational level. Analysis was based on the Fermi's golden rule approximation model. In the 4p-5s band region a vibrational sequence ofv(') v(")=1-1, 2-2, and 3-3 were recorded and a number of transitions belonging to the strongest (Q)Q(f(3)e) form branch of the 1-1 band were analyzed. PMID- 15836223 TI - Slow dynamics of supercooled m-toluidine investigated by mechanical spectroscopy. AB - Dynamics of supercooled m-toluidine close to the glass transition have been investigated by dynamic shear modulus measurements and stress relaxation experiments. The viscoelastic response of this material follows time-temperature superposition in the temperature range investigated. Comparison with results at ultrasonic frequencies suggests the existence of a secondary relaxation. A change of the temperature dependent viscosity from a Vogel-Fulcher-Tammann behavior to another regime at low temperatures is also found. Compared to most inorganic glass formers, the viscosity of m-toluidine at the glass transition is approximately two orders of magnitude lower. The shear relaxation times are characterized by the same temperature dependence as the viscosity. They are in reasonable agreement with the results of previous ultrasonic measurements. The conclusions of the present work agree with recent results obtained by high resolution dielectric spectroscopy. PMID- 15836224 TI - Reaction path determination for quantum mechanical/molecular mechanical modeling of enzyme reactions by combining first order and second order "chain-of-replicas" methods. AB - A two-step procedure for the determination of reaction paths in enzyme systems is presented. This procedure combines two chain-of-states methods: a quantum mechanical/molecular mechanical (QM/MM) implementation of the nudged elastic band (NEB) method and a second order parallel path optimizer method both recently developed in our laboratory. In the first step, a reaction path determination is performed with the NEB method, along with a restrained minimization procedure for the MM environment to obtain a first approximation to the reaction path. In the second step, the calculated path is refined with the parallel path optimizer method. By combining these two methods the reaction paths are determined accurately, and in addition, the number of path optimization iterations are significantly reduced. This procedure is tested by calculating both steps of the isomerization of 2-oxo-4-hexenedioate by 4-oxalocrotonate tautomerase, which have been previously determined by our group. The calculated paths agree with the previously reported results and we obtain a reduction of 45%-55% in the number of path optimization cycles. PMID- 15836225 TI - Computing the Soret coefficient in aqueous mixtures using boundary driven nonequilibrium molecular dynamics. AB - We have computed the Soret coefficient in aqueous mixtures using a boundary driven nonequilibrium molecular dynamics algorithm and standard molecular force fields. The choice of this specific approach is justified by the nature of the mixtures studied here. Four aqueous solutions, including methanol, ethanol, acetone, and dimethyl-sulfoxide (DMSO) have been studied at ambient conditions for different compositions. The experimental behavior of water-alcohol mixtures was reproduced, including the change of sign of the Soret coefficient with composition, in excellent agreement with existing experimental data. The methodology has been applied to obtain pure predictions for water-acetone and water-DMSO where no experimental data are accessible. A change of sign is also observed in the same range of composition as in water-alcohol mixtures. It is suggested that the nature and strength of the molecular interactions, rather than the mass or shape ratio of the components, dominates the behavior of the Soret coefficient versus composition for the aqueous associating mixtures studied here. PMID- 15836226 TI - Path integral ground state with a fourth-order propagator: application to condensed helium. AB - Ground state properties of condensed helium are calculated using the path integral ground state (PIGS) method. A fourth-order approximation is used as short (imaginary) time propagator. We compare our results with those obtained with other quantum Monte Carlo (QMC) techniques and different propagators. For this particular application, we find that the fourth-order propagator performs comparably to the pair product approximation, and is far superior to the primitive approximation. Results obtained for the equation of state of condensed helium show that PIGS compares favorably to other QMC methods traditionally utilized for this type of calculation. PMID- 15836227 TI - Role of hydrogen bonds in the fast dynamics of binary glasses of trehalose and glycerol: a molecular dynamics simulation study. AB - Trehalose-glycerol mixtures are known to be effective in the long time preservation of proteins. However, the microscopic mechanism of their effective preservation abilities remains unclear. In this article we present a molecular dynamics simulation study of the short time, less than 1 ns, dynamics of four trehalose-glycerol mixtures at temperatures below the glass transition temperature. We found that a mixture of 5% glycerol and 95% trehalose has the most suppressed short time dynamics (fast dynamics). This result agrees with the experimental analysis of the mean-square displacement of the hydrogen atoms, as measured via neutron scattering, and correlates with the experimentally observed enhancement of the stability of some enzymes at this particular concentration. Our microscopic analysis suggests that the formation of a robust intermolecular hydrogen bonding network is most effective at this concentration and is the main mechanism for the suppression of the fast dynamics. PMID- 15836228 TI - Moments of the rate of nonstationary nucleation. AB - The zeroth, first, and reduced moments of the rate of nonstationary nucleation at constant supersaturation are parameters in the description of the variation of this rate with time. The dependence of the reduced moment M(r) on the nucleus size n(*) and the supersaturation Deltamu is analyzed and it is found that under typical conditions this moment decreases from about 1.9 to 1.1 with increasing n(*) or, alternatively, decreasing Deltamu. Expressions for the M(r)(n(*)) and M(r)(Deltamu) dependences are given which allow approximate calculation of the reduced moment in different cases of one-component nucleation. PMID- 15836229 TI - The melting temperature of the most common models of water. AB - The melting temperature of ice I(h) for several commonly used models of water (SPC, SPC/E,TIP3P,TIP4P, TIP4P/Ew, and TIP5P) is obtained from computer simulations at p = 1 bar. Since the melting temperature of ice I(h) for the TIP4P model is now known [E. Sanz, C. Vega, J. L. F. Abascal, and L. G. MacDowell, Phys. Rev. Lett. 92, 255701 (2004)], it is possible to use the Gibbs-Duhem methodology [D. Kofke, J. Chem. Phys. 98, 4149 (1993)] to evaluate the melting temperature of ice I(h) for other potential models of water. We have found that the melting temperatures of ice I(h) for SPC, SPC/E, TIP3P, TIP4P, TIP4P/Ew, and TIP5P models are T = 190 K, 215 K, 146 K, 232 K, 245 K, and 274 K, respectively. The relative stability of ice I(h) with respect to ice II for these models has also been considered. It turns out that for SPC, SPC/E, TIP3P, and TIP5P the stable phase at the normal melting point is ice II (so that ice I(h) is not a thermodynamically stable phase for these models). For TIP4P and TIP4P/Ew, ice I(h) is the stable solid phase at the standard melting point. The location of the negative charge along the H-O-H bisector appears as a critical factor in the determination of the relative stability between the I(h) and II ice forms. The methodology proposed in this paper can be used to investigate the effect upon a coexistence line due to a change in the potential parameters. PMID- 15836230 TI - On the coupling between molecular diffusion and solvation shell exchange. AB - The connection between diffusion and solvent exchanges between first and second solvation shells is studied by means of molecular dynamics simulations and analytic calculations, with detailed illustrations for water exchange for the Li(+) and Na(+) ions, and for liquid argon. First, two methods are proposed which allow, by means of simulation, to extract the quantitative speed-up in diffusion induced by the exchange events. Second, it is shown by simple kinematic considerations that the instantaneous velocity of the solute conditions to a considerable extent the character of the exchanges. Analytic formulas are derived which quantitatively estimate this effect, and which are of general applicability to molecular diffusion in any thermal fluid. Despite the simplicity of the kinematic considerations, they are shown to well describe many aspects of solvent exchange/diffusion coupling features for nontrivial systems. PMID- 15836231 TI - A new force field for atomistic simulations of aqueous tertiary butanol solutions. AB - We present a new tert-butanol force field parametrized to reproduce the mixture thermodynamics of tert-butanol/water over a wide range of solution compositions at room temperature and atmospheric pressure. The experimental Kirkwood-Buff integrals, which quantify preferential solvation of solution components by the same species or by the other components, were used as target values to be reproduced. Water was modeled using the simple point charge model. In the range of alcohol mole fractions between 0.02 and 0.98, our optimized model satisfactorily reproduces alcohol-alcohol, water-water, and alcohol-water aggregation behavior. As a consequence, the solution activity derivatives are reproduced as well. A comparison has been made with solution activities obtained by free energy calculations (i.e., thermodynamic integration). It clearly shows that the Kirkwood-Buff based approach performs superior in predicting solution activities of liquid mixtures. The new tert-butanol model has been used to examine the solution structure and hydrophobic interactions in aqueous tert butanol at the various mixture compositions. A comparison is made with structural data obtained by neutron diffraction. PMID- 15836232 TI - Moderately and strongly supercooled liquids: a temperature-derivative study of the primary relaxation time scale. AB - The primary relaxation time scale tau(T) derived from the glass forming supercooled liquids (SCLs) is discussed within ergodic-cluster Gaussian statistics, theoretically justified near and above the glass-transformation temperature T(g). An analysis is given for the temperature-derivative data by Stickel et al. on the steepness and the curvature of tau(T). Near the mode coupling-theory (MCT) crossover T(c), these derivatives separate by a kink and a jump, respectively, the moderately and strongly SCL states. After accounting for the kink and the jump, the steepness remains a piecewise conitnuous function, a material-independent equation for the three fundamental characteristic temperatures, T(g), T(c), and the Vogel-Fulcher-Tamman (VFT) T(0), is found. Both states are described within the heterostructured model of solidlike clusters parametrized in a self-consistent manner by a minimum set of observable parameters: the fragility index, the MCT slowing-down exponent, and the chemical excess potential of Adam and Gibbs model (AGM). Below the Arrhenius temperature, the dynamically and thermodynamically stabilized clusters emerge with a size of around of seven to nine and two to three molecules above and close to T(g) and T(c), respectively. On cooling, the main transformation of the moderately into the strongly supercooled state is due to rebuilding of the cluster structure, and is attributed to its rigidity, introduced through the cluster compressibility. It is shown that the validity of the dynamic AGM (dynamically equivalent to the standard VFT form) is limited by the strongly supercooled state (T(g) < T < T(c)) where the superrigid cooperative rearranging regions are shown to be well-chosen parametrized solidlike clusters. Extension of the basic parameter set by the observable kinetic and diffusive exponents results in prediction of a subdiffusion relaxation regime in SCLs that is distinct from that established for amorphous polymers. PMID- 15836233 TI - Effective conjugation and Raman intensities in oligo(para-phenylene)s: a microscopic view from first-principles calculations. AB - Electron-phonon coupling in oligo(para-phenylene)s is addressed in terms of the off-resonance Raman intensities of two distinct modes at 1220 and 1280 cm(-1). On the basis of Albrecht's theory, vibrational coupling and Raman intensities are calculated from first-principles quantum-chemical methods. A few-state model is used to evaluate the dependence of the mode intensities on oligomer length, planarity, and excitation wavelength. The link between electron delocalizationconjugation and Raman intensities is highlighted. Extending on prior studies, the present work focuses on providing an in-depth understanding of the origin of this correlation in addition to reproducing experimental findings. The model applied here allows us to interpret the results on a microscopic, quantum-mechanical basis and to relate the observed trends to the molecular orbital structure and nature of the excited states in this class of materials. We find quantitative agreement between the results of the calculations and those of measurements performed on oligo(para-phenylene)s of various chain lengths in the solid state and in solution. PMID- 15836234 TI - Ab initio molecular dynamics of liquid hydrogen chloride. AB - We carried out an ab initio molecular dynamics simulation of liquid hydrogen chloride (l-HCl) at a temperature of 313 K. Comparison with inelastic neutron scattering data shows that the simulation achieves an overall good description of the structural correlations, improving significantly upon a description based on classical interaction potentials. Despite some minor differences between theory and experiment in the H-H partial structure factor, the simulation gives a description of the hydrogen bonding in impressive agreement with experiment, for both the amount and the bond-length distribution of the bonds. In the simulation, 40% of the molecules are nonbonded, while the hydrogen-bonded chains are short, principally consisting of dimers (25%) and trimers (15%). Neighboring molecules in the simulation are found to form L-shaped arrangements, like in the isolated (HCl)(2) dimer and in crystalline phases of HCl. The time correlation of the molecular-axis orientation is found to be characterized by a very short decay time (0.13 ps), consistent with the short length of the hydrogen-bonded chains. Other dynamical properties investigated in this work include the diffusion coefficient and the vibrational density of states. We evaluated the molecular dipole of the HCl molecule in the liquid using a definition based on the coupling of rotational modes to an external electric field. The average dipole moment (1.53 D) derived in this way is found to be considerably larger than for the isolated molecule (1.11 D). Our results show that the dipole moment in [script-l] HCl undergoes large fluctuations, both in orientation and in modulus. Upon the onset of an external field, such dipole fluctuations concur to reduce the fluctuations of the dielectric response. PMID- 15836235 TI - Effects of pulsed low frequency electromagnetic fields on water using photoluminescence spectroscopy: role of bubble/water interface. AB - The effects of a pulsed low frequency electromagnetic field were investigated on photoluminescence of well characterized water and prepared under controlled conditions (container, atmospheric, electromagnetic, and acoustic environments). When reference water samples were excited at 260 nm, two wide emission bands centered at 345 nm (3.6 eV) and 425 nm (2.9 eV) were observed. By contrast under 310 nm excitation, only one band appeared at 425 nm. Interestingly, electromagnetic treatment (EMT) induced, at both excitation wavelengths, a decrease (around 70%) in the 425 nm band relative photoluminescence intensity. However, no difference between reference and treated sample was observed in the 345 nm band. Other experiments, performed on outgassed samples (reference and treated), show that the emission bands (position, shape, intensity) under excitation at 260 nm and 310 nm were similar and close to the corresponding bands of the treated nonoutgassed samples. Similar effects were observed on photoluminescence excitation of water samples. Two excitation bands monitored at 425 nm were observed at 272 nm and 330 nm. After EMT and/or outgassing, a decrease (>60%) was observed in the intensity of these two bands. Altogether, these results indicate that electromagnetic treatment and/or outgassing decrease in a similar fashion the photoluminescence intensity in water samples. They also suggest that this effect is most likely indirectly attributed to the presence of gas bubbles in water. The possible role of hydrated ionic shell around the bubbles in the observed extraluminescence is discussed. PMID- 15836236 TI - Transient complex oscillations in a closed chemical system with coupled autocatalysis. AB - In this study, hydroquinone was introduced to the classic Belousov-Zhabotinsky (BZ) reaction to build up coupled autocatalytic feedbacks. Various complex dynamical behaviors including successive period-adding bifurcations, irregular oscillations, and frequency modulations were observed in the coupled reaction system. Not only the complexity of oscillations but also the time period during which complex oscillations persist were found to depend greatly on the initial concentration of hydroquinone, which was expected to manifest the coupling strength in the studied system. Dependence of the observed transient complex oscillations on concentrations of ferroin, sulfuric acid, bromate, and malonic acid was also characterized systematically. Numerical simulations with a modified BZ model via incorporating reactions involving hydroquinone and products of hydroquinone qualitatively reproduced the influence of hydroquinone seen in experiments. PMID- 15836237 TI - Vibronic fine structure in the absorption spectrum of oligothiophene thin films. AB - A multimode Holstein Hamiltonian is used to describe optical excitations in quaterthiophene pinwheel aggregates. The Hamiltonian includes the coupling of excitons originating from the 1A(g)-->1B(u) electronic transition to phonons originating from the five intramolecular vibrational modes known from oligothiophene solution absorption/emission spectroscopy. The resulting eigenstates with lowest energy are best described as hybrid polaron phonons. The polarons are formed by coupling excitons with the higher frequency (688, 1235, and 1551 cm(-1)) vibrational modes, while the (optical) phonons arise from the lower frequency (161 and 333 cm(-1)) modes. The polaron phonons are responsible for the fine structure defining the A(1) band in the low-energy region of the absorption spectrum, ranging from the band origin to approximately 1500 cm(-1) beyond. The calculated A(1) band of quaterthiophene aggregates agrees favorably with that observed from thin films. PMID- 15836238 TI - Rotationally inelastic scattering of HD from Cu(100) and Pd(111). AB - Rotational excitation of HD scattered from Cu(100), Pd(111), and Pd(111):H(D) was measured using molecular beam and quantum-state-specific laser spectroscopy techniques. Greater than 91% of the incident HD population was in the v = 0, J = 0 state. The final rotational distributions from Cu(100), Pd(111), and Pd(111):H(D) were compared for a HD beam at an incident energy of 74 meV. For all the three surfaces studied, rotationally inelastic scattering probabilities were large. We find that the final HD rotational distributions are remarkably similar for the three surfaces even though Pd(111) is very reactive to dissociative adsorption of HD whereas Cu(100) and Pd(111):H(D) are chemically inert. PMID- 15836239 TI - Adsorption of multiblock copolymers onto a chemically heterogeneous surface: a model of pattern recognition. AB - We present a statistical mechanical model, which is used to investigate the adsorption behavior of two-letter (AB) copolymers on chemically heterogeneous surfaces. The surfaces with regularly distributed stripes of two types (A and B) and periodic multiblock copolymers (Al)B(l))(x) are studied. It is assumed that A(B)-type segments selectively adsorb onto A(B)-type stripes. It is shown that the adsorption strongly depends on the copolymer sequence distribution and the arrangement of selectively adsorbing regions on the surface. The polymer-surface binding proceeds as a two-step process. At the first step, the copolymer having short blocks adsorbs onto the surface as an effective homopolymer, which does not feel chemical pattern. At the second step, when the polymer-surface attraction is sufficiently strong, the adsorbed chain adjusts its equilibrium conformation to reach the perfect bound state, thereby demonstrating ability for pattern recognition. The key element of this mechanism is the redistribution of strongly adsorbed copolymer diblocks A(l)B(l), which behave as surfactants, between multiple AB interfaces separating A and B stripes on the adsorbing surface. Such redistribution is accompanied by a well-pronounced decrease in the system entropy. We have found that marked pattern recognition is possible for copolymers with relatively short blocks at high polymer/surface affinities, beyond the adsorption threshold. PMID- 15836240 TI - Fluorescence lifetime fluctuations of single molecules probe local density fluctuations in disordered media: a bulk approach. AB - We investigated the nanometer scale mobility of polymers in the glassy state by monitoring the dynamics of embedded single fluorophores. Recently we reported on fluorescence lifetime fluctuations which reflect the segmental rearrangement dynamics of the polymer in the surroundings of the single molecule probe. Here we focus on the nature of these fluorescence lifetime fluctuations. First the potential role of quenching and molecular conformational changes is discussed. Next we concentrate on the influence of the radiative density of states on the spontaneous emission of individual dye molecules embedded in a polymer. To this end we present a theory connecting the effective-medium theory to a cell-hole model, originating from the Simha-Somcynsky free-volume theory. The relation between the derived distributions of free volume and fluorescence lifetime allows one to determine the number of segments involved in the local rearrangement directly from experimental data. Results for two different polymers as a function of temperature are presented. PMID- 15836241 TI - Exact sequence analysis for three-dimensional hydrophobic-polar lattice proteins. AB - We have exactly enumerated all sequences and conformations of hydrophobic-polar (HP) proteins with chains of up to 19 monomers on the simple cubic lattice. For two variants of the HP model, where only two types of monomers are distinguished, we determined and statistically analyzed designing sequences, i.e., sequences that have a nondegenerate ground state. Furthermore we were interested in characteristic thermodynamic properties of HP proteins with designing sequences. In order to be able to perform these exact studies, we applied an efficient enumeration method based on contact sets. PMID- 15836242 TI - Controlling the size of nanostructures in thin films via blending of block copolymers and homopolymers. AB - The effects of molecular weight and concentration of poly (methyl methacrylate) (PMMA) homopolymer or symmetric short polystyrene-block-poly (methyl methacrylate) (PS-b-PMMA) diblock copolymer on the size of the nanostructures of its blends with symmetric long PS-b-PMMA diblock copolymer have been investigated by atomic force microscopy. By careful controlling of the film thickness, solvent selectivity, and annealing time, PMMA cylindrical microdomains oriented normal to the film surface were obtained in all thin films. With the addition of both low- and high-molecular-weight PMMA homopolymers, the cylindrical domain sizes increased although it was less obvious for the lower molecular weight homopolymer. In contrast to the homopolymer, adding the short chain diblock copolymer resulted in a decrease in the cylindrical domain size, which was ascribed to the reduction of the interfacial tension and increase in the stretching energy. PMID- 15836243 TI - Second-harmonic generation optical activity of a polypeptide alpha-helix at the air/water interface. AB - Quantitative measurements of second-harmonic generation optical activity (SHG-OA) have been performed for alpha-helical polypeptides poly-(gamma-benzyl-L glutamate) and poly-(gamma-ethyl-L-glutamate) adsorbed at the airwater interface, with the fundamental frequency variant Planck's over 2piomega = 2.96 eV (lambda = 417 nm). The chiral component of the nonlinear susceptibility chi(XYZ) ((2)) is small for both polymers, being comparable in magnitude with the susceptibility chi(XXZ) ((2)) of the clean airwater interface. The microscopic origin of the nonlinear response has been investigated by using semiempirical ZINDOS calculations in conjunction with standard time-dependent perturbation theory to evaluate the molecular hyperpolarizability tensor of a model alpha-helix composed of glycine residues. Calculated nonlinear susceptibilities (per monomer unit) are in good agreement with experimental measurements for both the chiral and achiral response. The computational results indicate that charge transfer transitions of the alpha-helix have a large influence on the achiral components of the hyperpolarizability tensor, and produce characteristic features in the response under suitable experimental conditions. The dominant origin of SHG-OA for the model alpha-helix is a structural effect due to the tilt of the plane of each amide group of the helix relative to the helical axis. SHG-OA is associated with the orientational distribution of isolated, achiral chromophores, and is present in the absence of electronic coupling between the amide subunits of the polypeptide alpha-helix. PMID- 15836244 TI - Single wavelength excitation fluorescence cross-correlation spectroscopy with spectrally similar fluorophores: resolution for binding studies. AB - It was shown recently that fluorescence cross-correlation spectroscopy (FCCS) can be performed using a single laser wavelength for excitation (SW-FCCS) [L. C. Hwang and T. Wohland, Chem. Phys. Chem 5, 549 (2004).]. This method simplifies the FCCS setup since it does not require the simultaneous alignment of two lasers to the same focal spot. But up to now the method was shown to work only with dyes possessing large Stokes' shifts, and thus was limited to the use of quantum dots and tandem dyes. In this work we show that standard organic dyes with overlapping emission spectra, for instance fluorescein and tetramethylrhodamine, can be used as fluorescent pairs in SW-FCCS. As a biological model system for ligand-receptor interactions we studied the binding of biotin to streptavidin. To investigate the applicability of SW-FCCS for binding studies we adapt the existing FCCS theory for SW-FCCS and calculate limits for the measurement of dissociation constants in dependence on sample concentration, sample purity, and spectral cross talk between the different detection channels. PMID- 15836245 TI - The molecular approach to heterogeneous nucleation. AB - A molecular approach to heterogeneous nucleation has been developed. The expressions for the equilibrium cluster distribution, the reversible work of the cluster formation, and the nucleation rate have been derived. Two separate statements for the work of formation were formulated. If the equilibrium cluster distribution is normalized on the monomer concentration near the substrate surface, the reversible work of formation is expressed by DeltaG(het) (I) = (F(n) (het)-F(n) (hom))-(F(1) (het)-F(1) (hom)) + DeltaG(hom) where F(n) (het) and F(n) (hom) are the Helmholtz free energies of a cluster interacting with a substrate and a cluster not interacting with the substrate, respectively. If the equilibrium cluster distribution is normalized on the monomer concentration far from the substrate surface, the work of cluster formation is given by DeltaG(het) (II) = (F(n) (het)-F(n) (hom)) + DeltaG(hom). The former expression corresponds to the approach of the classical heterogeneous nucleation theory. The cluster partition function appears to be dependent on the location of a virtual plane, which separates the volume, where the interaction of the clusters with the substrate is effective from the one where interaction is negligible. Our Monte Carlo simulations have shown that the dependence is rather weak and thus the location of the plane is not very important. According to the simulations the variation of the plane position in the range from 20 to 50 Angstroms does not lead to a considerable change of the heterogeneous nucleation rate. PMID- 15836246 TI - Stochastic chemical reactions in microdomains. AB - Traditional chemical kinetics may be inappropriate to describe chemical reactions in microdomains involving only a small number of substrate and reactant molecules. Starting with the stochastic dynamics of the molecules, we derive a master-diffusion equation for the joint probability density of a mobile reactant and the number of bound substrate in a confined domain. We use the equation to calculate the fluctuations in the number of bound substrate molecules as a function of initial reactant distribution. A second model is presented based on a Markov description of the binding and unbinding and on the mean first passage time of a molecule to a small portion of the boundary. These models can be used for the description of noise due to gating of ionic channels by random binding and unbinding of ligands in biological sensor cells, such as olfactory cilia, photoreceptors, hair cells in the cochlea. PMID- 15836247 TI - Dielectric permittivity profiles of confined polar fluids. AB - The dielectric response of a simple model of a polar fluid near neutral interfaces is examined by a combination of linear response theory and extensive molecular dynamics simulations. Fluctuation expressions for a local permittivity tensor epsilon(r) are derived for planar and spherical geometries, based on the assumption of a purely local relationship between polarization and electric field. While the longitudinal component of epsilon exhibits strong oscillations on the molecular scale near interfaces, the transverse component becomes ill defined and unphysical, indicating nonlocality in the dielectric response. Both components go over to the correct bulk permittivity beyond a few molecular diameters. Upon approaching interfaces from the bulk, the permittivity tends to increase, rather than decrease as commonly assumed, and this behavior is confirmed for a simple model of water near a hydrophobic surface. An unexpected finding of the present analysis is the formation of "electrostatic double layers" signaled by a dramatic overscreening of an externally applied field inside the polar fluid close to an interface. The local electric field is of opposite sign to the external field and of significantly larger amplitude within the first layer of polar molecules. PMID- 15836248 TI - Glycine-ice nanolayers: morphology and surface energetics. AB - Ultrathin glycine-ice films (nanolayers) have been prepared in ultrahigh vacuum by condensation of H(2)O and glycine at 110 K and 150 K on single crystalline Al(2)O(3) surfaces and have been investigated by temperature programed thermal desorption, x-ray photoelectron spectroscopy, and work function measurements. Various layer architectures have been considered, including glycine-on-ice, ice on-glycine, and mixed glycine-ice nanolayers. Low coverages of adsorbed glycine molecules on amorphous ice surfaces suppress the amorphous-to-crystalline phase transition in the temperature range 140-160 K in near-surface regions and consequently lead to a lower desorption temperature of H(2)O molecules than from pure ice layers. Thicker glycine overlayers on ice provide a kinetic restriction to H(2)O desorption from the underlying ice layers until the glycine molecules become mobile and develop pathways for water desorption at higher temperature (>170 K). Ice overlayers do not wet glycine film surfaces, but the glycine molecules on ice are sufficiently immobile at 110 K, so that continuous glycine overlayers form. In mixed glycine-ice nanolayers the glycine phase displays hydrophobic behavior and a phase separation takes place, with the accumulation of glycine near the surfaces of the films. PMID- 15836249 TI - Variations of surfactant monolayer surface density in induced steady wave regime. AB - Fluctuations of an insoluble surfactant concentration along the free liquid surface induced by steady surface waves are considered theoretically. The energy of a waved surface is assumed to consist of surface tension, curvature, and van der Waals energy components. Dependencies of the surface tension and the bending stiffness versus the surfactant concentration are assumed to be linear relative to some reference level. The van der Waals energy is taken in the form of interaction term for a thin film. Minimization of the total energy allows the expression for the deviations of concentration to be obtained. The distribution of a surfactant concentration relative to some reference level has been found to be periodic, with a period that is half of the wave period, and the amplitude of oscillations is a function of a wave number that is very similar to the Landau expansion of the free-energy near the critical point in phase transitions. PMID- 15836250 TI - Excitation of the shear horizontal mode in a monolayer by inelastic helium atom scattering. AB - Inelastic scattering of a low-energy atomic helium beam (HAS) by a physisorbed monolayer is treated in the one-phonon approximation using a time-dependent wave packet formulation. The calculations show that modes with shear horizontal polarization can be excited near high symmetry azimuths of the monolayer, in agreement with recent experiments. The parameters of the calculations are chosen to match the conditions of HAS experiments for triangular incommensurate monolayer solids of xenon, krypton, and argon adsorbed on the (111) face of platinum, and the results show many of the systematic experimental trends for relative excitation probability of the shear horizontal and longitudinal acoustic phonon branches. The inelastic scattering at beam energies near 8 meV is exceedingly sensitive to small misalignment between the scattering plane and the high symmetry directions of the monolayer solid. The diffraction and inelastic processes arise from a strong coupling of the incident atom to the target and the calculated results show large departures from expectations based on analogies to inelastic thermal neutron scattering. PMID- 15836251 TI - The equilibrium properties and folding kinetics of an all-atom Go model of the Trp-cage. AB - The ultrafast-folding 20-residue Trp-cage protein is quickly becoming a new benchmark for molecular dynamics studies. Already several all-atom simulations have probed its equilibrium and kinetic properties. In this work an all-atom Go model is used to accurately represent the side-chain packing and native atomic contacts of the Trp-cage. The model reproduces the hallmark thermodynamics cooperativity of small proteins. Folding simulations observe that in the fast folding dominant pathway, partial alpha-helical structure forms before hydrophobic core collapse. In the slow-folding secondary pathway, partial core collapse occurs before helical structure. The slow-folding rate of the secondary pathway is attributed to the loss of side-chain rotational freedom, due to the early core collapse, which impedes the helix formation. A major finding is the observation of a low-temperature kinetic intermediate stabilized by a salt bridge between residues Asp-9 and Arg-16. Similar observations [R. Zhou, Proc. Natl. Acad. Sci. U.S.A. 100, 13280 (2003)] were reported in a recent study using an all atom model of the Trp-cage in explicit water, in which the salt-bridge stabilized intermediate was hypothesized to be the origin of the ultrafast-folding mechanism. A theoretical mutation that eliminates the Asp-9-Arg-16 salt bridge, but leaves the residues intact, is performed. Folding simulations of the mutant Trp-cage observe a two-state free-energy landscape with no kinetic intermediate and a significant decrease in the folding rate, in support of the hypothesis. PMID- 15836252 TI - The dynamics of single chains within a model polymer melt. AB - Discontinuous molecular dynamics simulations are performed on a system containing 32 hard chains of length 192 at a volume fraction of phi = 0.45 to explore the idea that localized entanglements have a significant effect on the dynamics of the individual chains within an entangled polymer melt. Anomalous behavior can still be observed when studying the dynamics of the individual chains, although increased time averaging causes the anomalous relaxation-memory-release behavior that was observed previously in the system to smooth out. First, the individual chain mean squared displacements and apparent diffusion coefficients are calculated, and a wide distribution of diffusive behavior is found. Although the apparent diffusion coefficient curve averaged over all chains displays the predicted long-time diffusive behavior, the curves for the individual chains differ both qualitatively and quantitatively. They display superdiffusive, diffusive, and subdiffusive behavior, with the largest percentage of chains exhibiting superdiffusive behavior and the smallest percentage exhibiting the predicted diffusive behavior. Next, the individual chain end-to-end vector autocorrelation functions and relaxation times are determined, and a wide distribution of stress relaxation behavior is found. The times when the end-to end vector autocorrelation functions relax completely span almost an order of magnitude in reduced time. For some chains, the end-to-end vector autocorrelation function relaxes smoothly toward zero similar to the system average; however, for other chains the relaxation is slowed greatly, indicating the presence of additional entanglements. Almost half of the chains exhibit the anomalous behavior in the end-to-end vector autocorrelation function. Finally, the dynamic properties are displayed for a single chain exhibiting anomalous relaxation memory-release behavior, supporting the idea that the relaxation-memory-release behavior is a single-chain property. PMID- 15836253 TI - Simulation of conformational transitions by the restricted perturbation-targeted molecular dynamics method. AB - A method for the simulation of conformational transitions is presented. The method, based on targeted molecular dynamics (TMD), limits the conformational change at each molecular dynamics step to a fixed size, that minimizes the root mean square deviation from the target. The method is more efficient than standard TMD and yields lower energy pathways, but, like the TMD method, requires only a single molecular dynamics simulation. Test calculations and comparisons with standard TMD calculations for the alanine dipeptide with the analytic continuum electrostatics implicit solvent model are presented. PMID- 15836254 TI - Stretching a heteropolymer. AB - We study the elastic properties of single heteropolymers. By means of exact enumeration of conformations, Monte Carlo (MC) simulation, and variational principles, we calculate equilibrium force-extension curves of heterocopolymers for specific arrangements of the monomer types along the sequence. At a given extension z, the time averaged measured force is the weighted sum of restoring forces for various configurations. Using variational principles, we calculate force-extension (f-z) curves of heteropolymers with fixed extensions z. These results are compared with f-z curves obtained from MC simulations and exact enumeration of all conformations. Typical random sequences manifest several piecewise unfoldings of blocks of various size, which are overlapping due to thermal fluctuations. The shape of the elastic response of a heteropolymer reflects the disorder in the primary block structure and the binding energies of these blocks. PMID- 15836255 TI - A new scheme to generalized (lag, anticipated, and complete) synchronization in chaotic and hyperchaotic systems. AB - In this paper, a generalized (lag, anticipated, and complete) synchronization of a class of continuous-time systems is defined. A systematic, powerful and concrete scheme is developed to investigate the generalized (lag, anticipated, and complete) synchronization between the drive system and response system based on the active control idea. The hyperchaotic Rossler system, transformed Rossler and Chen system as well as two coupled nonidentical Rossler oscillators are chosen to illustrate the proposed scheme. Numerical simulations are used to verify the effectiveness of the proposed scheme. The scheme can be also extended to research generalized (lag, anticipated, and complete) synchronization between other dynamical systems. PMID- 15836256 TI - Experimental observation of Lorenz chaos in the Quincke rotor dynamics. AB - In this paper, we report experimental evidence of Lorenz chaos for the Quincke rotor dynamics. We study the angular motion of an insulating cylinder immersed in slightly conducting oil and submitted to a direct current electric field. The simple equations which describe the dynamics of the rotor are shown to be equivalent to the Lorenz equations. In particular, we observe two bifurcations in our experimental system. Above a critical value of the electric field, the cylinder rotates at a constant rate. At a second bifurcation, the system becomes chaotic. The characteristic shape of the experimental first return map provides strong evidence for Lorenz-type chaos. PMID- 15836257 TI - Coupling-mediated ghost resonance in mutually injected lasers. AB - We experimentally and numerically study the phenomenon of ghost resonance in coupled nonlinear systems. Two mutually injected semiconductor lasers are externally perturbed in their pump currents by two respective periodic signals of different frequencies f(1) and f(2). For small amplitudes of the external modulations, the two laser intensities display synchronized optical pulses, in the form of dropout events occurring at irregular times. By adjusting the amplitude and frequencies of the driving signals, the system exhibits a ghost resonance in the dropout appearance at a frequency f(r) not present in the distributed inputs. PMID- 15836258 TI - On chaotic behavior of gravitating stellar shells. AB - Motion of two gravitating spherical stellar shells around a massive central body is considered. Each shell consists of point particles with the same specific angular momenta and energies. In the case when one can neglect the influence of gravitation of one ("light") shell onto another ("heavy") shell ("restricted problem") the structure of the phase space is described. The scaling laws for the measure of the domain of chaotic motion and for the minimal energy of the light shell sufficient for its escape to infinity are obtained. PMID- 15836259 TI - Dynamic characterization of hysteresis elements in mechanical systems. I. Theoretical analysis. AB - The pre-sliding-pre-rolling phase of friction behavior is dominated by rate independent hysteresis. Many machine elements in common engineering use exhibit, therefore, the characteristic of "hysteresis springs," for small displacements at least. Plain and rolling element bearings that are widely used in motion guidance of machine tools are typical examples. While the presence of a hysteresis element may mark the character of the resulting dynamics, little is to be found about this topic in the literature. The study of the nonlinear dynamics caused by such elements becomes imperative if we wish to achieve accurate control of such machines. In this Part I of the investigation, we examine a single-degree-of freedom mass-hysteresis-spring system and show that, while the free response case is amenable to an exact solution, the more important case of forced response has no closed form solution and requires other methods of treatment. We consider harmonic-balance analysis methods (which are common analysis tools in engineering) suitable for frequency-domain treatment, in particular the approximate describing function (DF) method, and compare those results with "exact" numerical simulations. The DF method yields basically a linear equation with amplitude-dependent modal parameters. We find that agreement in the frequency response function, between DF and exact solution, is good for small excitation amplitudes and for very large amplitudes. Intermediate values, however, show high sensitivity to amplitude variations and, consequently, no regular solution is obtainable by either approach. This appears to be an inherent property of the system pointing to the need for developing further analysis methods. Experimental verification of the analysis outlined in this Part I is given in Part II of the paper. PMID- 15836260 TI - Dynamic characterization of hysteresis elements in mechanical systems. II. Experimental validation. AB - The industrial demand for machine tools with ever increasing speed and accuracy calls for a closer look at the physical phenomena that are present at small movements of those machine's slides. One of these phenomena, and probably the most dominant one, is the dependence of the friction force on displacement that can be described by a rate-independent hysteresis function with nonlocal memory. The influence of this highly nonlinear effect on the dynamics of the system has been theoretically analyzed in Part I of this paper. This part (II) aims at verifying these theoretical results on three experimental setups. Two setups, consisting of linearly driven rolling element guideways, have been built to specifically study the hysteretic friction behavior. The experiments performed on these specially designed setups are then repeated on one axis of an industrial pick-and-place device, driven by a linear motor and guided by commercial guideways. The results of the experiments on all the setups agree qualitatively well with the theoretically predicted ones and point to the inherent difficulty of accurate quantitative identification of the hysteretic behavior. They further show that the hysteretic friction behavior has a direct bearing on the dynamics of machine tools and its presence should therefore be carefully considered in the dynamic identification process of these systems. PMID- 15836261 TI - Hierarchy of bifurcations in the truncated and forced nonlinear Schrodinger model. AB - The truncated forced nonlinear Schrodinger (NLS) model is known to mimic well the forced NLS solutions in the regime at which only one linearly unstable mode exists. Using a novel framework in which a hierarchy of bifurcations is constructed, we analyze this truncated model and provide insights regarding its global structure and the type of instabilities which appear in it. In particular, the significant role of the forcing frequency is revealed and it is shown that a parabolic resonance mechanism of instability arises in the relevant parameter regime of this model. Numerical experiments demonstrating the different types of chaotic motion which appear in the model are provided. PMID- 15836262 TI - On Jacobian matrices for flows. AB - We present a general method for constructing numerical Jacobian matrices for flows discretized on a Poincare surface of section. Special attention is given to Hamiltonian flows where the additional constraint of energy conservation is explicitly taken into account. We demonstrate the approach for a conservative dynamical flow and apply the technique for the general detection of periodic orbits. PMID- 15836263 TI - Chaotic system for self-synchronizing Doppler measurement. AB - In a radar system, it is necessary to measure both range and velocity of a target. The movement of the target causes a Doppler shift of the radar signal, and the size of the Doppler shift is used to measure the velocity of the target. In this work, a chaotic drive-response system is simulated that detects a Doppler shift in a chaotic signal. The response system can detect Doppler shifts in more than one signal at a time. PMID- 15836264 TI - Robust discretizations versus increase of the time step for the Lorenz system. AB - When continuous systems are discretized, their solutions depend on the time step chosen a priori. Such solutions are not necessarily spurious in the sense that they can still correspond to a solution of the differential equations but with a displacement in the parameter space. Consequently, it is of great interest to obtain discrete equations which are robust even when the discretization time step is large. In this paper, different discretizations of the Lorenz system are discussed versus the values of the discretization time step. It is shown that the sets of difference equations proposed are more robust versus increases of the time step than conventional discretizations built with standard schemes such as the forward Euler, backward Euler, or centered finite difference schemes. The nonstandard schemes used here are Mickens' scheme and Monaco and Normand-Cyrot's scheme. PMID- 15836265 TI - Current oscillation and chaotic dynamics in superlattices driven by crossed electric and magnetic fields. AB - We have theoretically studied current oscillation and chaotic dynamics in doped GaAsAlAs superlattices driven by crossed electric and magnetic fields. When the superlattice system is driven by a dc voltage, a stationary or dynamic electric field domain can be obtained. We carefully studied the electric-field-domain dynamics and current self-oscillation which both display different modes with the change of magnetic field. When an ac electric field is also applied to the superlattice, a typical nonlinear dynamic system is constructed with the ac amplitude, ac frequency, and magnetic field as the control parameters. Different nonlinear behaviors show up when we tune the control parameters. PMID- 15836266 TI - Dissipative solitons driving and bound state control via parameter gradients. AB - The effect of phase and intensity gradients on the motion of dissipative solitons in a nonlinear interferometer is investigated. We show how the forces exerted by parameter gradients on the solitons alter their interactions. Consequently, it is possible to tune the spectrum of soliton bound states via the introduction of proper spatial distribution of the system parameters. Furthermore, we show how the use of properly shaped parameter distributions can lead to a relevant reduction of the cross-talk between close by solitons in applications. PMID- 15836267 TI - Modeling wave propagation in realistic heart geometries using the phase-field method. AB - We present a novel algorithm for modeling electrical wave propagation in anatomical models of the heart. The algorithm uses a phase-field approach that represents the boundaries between the heart muscle and the surrounding medium as a spatially diffuse interface of finite thickness. The chief advantage of this method is to automatically handle the boundary conditions of the voltage in complex geometries without the need to track the location of these boundaries explicitly. The algorithm is shown to converge accurately in nontrivial test geometries with no-flux (zero normal current) boundary conditions as the width of the diffuse interface becomes small compared to the width of the cardiac action potential wavefront. Moreover, the method is illustrated for anatomically realistic models of isolated rabbit and canine ventricles as well as human atria. PMID- 15836268 TI - Breaking of balanced and unbalanced equatorial waves. AB - A clear-cut signature of a wave-breaking event is irreversible modification of the mean flow. In this paper, we provide examples of different breaking mechanisms and show that breaking scenario of equatorial waves in the beta-plane shallow water model is determined by the degree of balance between the zonal component of the Coriolis force and the pressure gradient. Our analysis is based on a specially designed numerical method which guarantees two essential conditions to simulate nonlinear equatorial waves: (i) the scheme converges toward weak solutions including shocks and (ii) preserves the steadiness of balanced stationary solutions. This allows for accurate diagnostics of Lagrangian invariants of motion such as passive tracer density or potential vorticity. For unbalanced waves, the lack of balance leads to shock formation in finite time. In shock fronts, the variation of the dissipation rate induces a nonadvective potential vorticity flux and violates the local potential vorticity conservation valid for smooth solutions. This dissipative breaking mechanism is generic for unbalanced waves and is associated with enhanced mixing. For long, balanced (Rossby) waves, breaking consists in appearance of recirculation regions. It results in the formation of propagating patterns, the equatorial modons, which trap fluid particles. Such breaking occurs during the propagation of Rossby wave packets with positive geopotential anomaly and is strengthened by decreasing fluid depth. The modons are robust and collide quasielastically with Kelvin waves. PMID- 15836269 TI - Dispersive destabilization of nonlinear light propagation in fiber Bragg gratings. AB - The effect of retaining the material dispersion terms in the nonlinear coupled mode equations (NLCME) that describe light propagation in fiber Bragg gratings is analyzed. It is found that dispersion is responsible for new instabilities of the uniform states and gives rise to new complex spatio-temporal dynamics that is not captured by the standard NLCME formulation. A detailed analysis of the effect of dispersion on the linear stability characteristics of the uniform solutions is presented and some numerical integrations of the NLCME with dispersion are also performed in order to corroborate the theoretical results. PMID- 15836270 TI - Detecting synchronizations in an asymmetric vocal fold model from time series data. AB - A nonlinear modeling approach is presented for the reconstruction of the synchronization structure in an asymmetric two-mass model from time series data. The asymmetric two-mass model describes a variety of normal and pathological human voices associated with synchronous and desynchronous oscillations of the two asymmetric vocal folds. Our technique recovers the synchronization diagram, which yields the regimes of synchronization as well as desynchronization, which are dependent upon the asymmetry parameter and the subglottal pressure. This allows the prediction of the regime of pathological phonation associated with desynchronization of the vocal folds from a few sets of recorded time series. It is shown that the modeling is quite effective when the time series data are chaotic and if they are taken from a regime of desynchronization. We discuss the applicability of the present approach as a diagnostic tool for voice pathologies. PMID- 15836271 TI - Breaking a chaos-noise-based secure communication scheme. AB - This paper studies the security of a secure communication scheme based on two discrete-time intermittently chaotic systems synchronized via a common random driving signal. Some security defects of the scheme are revealed: 1) The key space can be remarkably reduced; 2) the decryption is insensitive to the mismatch of the secret key; 3) the key-generation process is insecure against known/chosen plaintext attacks. The first two defects mean that the scheme is not secure enough against brute-force attacks, and the third one means that an attacker can easily break the cryptosystem by approximately estimating the secret key once he has a chance to access a fragment of the generated keystream. Yet it remains to be clarified if intermittent chaos could be used for designing secure chaotic cryptosystems. PMID- 15836272 TI - Oscillator clustering in a resource distribution chain. AB - The paper investigates the special clustering phenomena that one can observe in systems of nonlinear oscillators that are coupled via a shared flow of primary resources (or a common power supply). This type of coupling, which appears to be quite frequent in nature, implies that one can no longer separate the inherent dynamics of the individual oscillator from the properties of the coupling network. Illustrated by examples from microbiological population dynamics, renal physiology, and electronic oscillator theory, we show how competition for primary resources in a resource distribution chain leads to a number of new generic phenomena, including partial synchronization, sliding of the synchronization region with the resource supply, and coupling-induced inhomogeneity. PMID- 15836273 TI - Chaotic synchronization of coupled electron-wave systems with backward waves. AB - The chaotic synchronization of two electron-wave media with interacting backward waves and cubic phase nonlinearity is investigated in the paper. To detect the chaotic synchronization regime we use a new approach, the so-called time scale synchronization [Chaos 14, 603-610 (2004)]. This approach is based on the consideration of the infinite set of chaotic signals' phases introduced by means of continuous wavelet transform. The complex space-time dynamics of the active media and mechanisms of the time scale synchronization appearance are considered. PMID- 15836274 TI - Hopping behavior in the Kuramoto-Sivashinsky equation. AB - We report the first observations of numerical "hopping" cellular flame patterns found in computer simulations of the Kuramoto-Sivashinsky equation. Hopping states are characterized by nonuniform rotations of a ring of cells, in which individual cells make abrupt changes in their angular positions while they rotate around the ring. Until now, these states have been observed only in experiments but not in truly two-dimensional computer simulations. A modal decomposition analysis of the simulated patterns, via the proper orthogonal decomposition, reveals spatio-temporal behavior in which the overall temporal dynamics is similar to that of equivalent experimental states but the spatial dynamics exhibits a few more features that are not seen in the experiments. Similarities in the temporal behavior and subtle differences in the spatial dynamics between numerical hopping states and their experimental counterparts are discussed in more detail. PMID- 15836275 TI - Chaotic systems that are robust to added noise. AB - While added noise can destroy synchronization in synchronized chaotic systems, it was shown that some chaotic systems were not sensitive to added noise. In this paper, the mechanism for this noise resistance is explored. It is seen that part of the chaotic system acts like it is resonant, reducing the noise sensitivity of the system. By comparing to a model of a neuron, it is speculated that similar mechanisms may also be present in biological systems. PMID- 15836276 TI - Does synchronization of networks of chaotic maps lead to control? AB - We consider networks of chaotic maps with different network topologies. In each case, they are coupled in such a way as to generate synchronized chaotic solutions. By using the methods of control of chaos we are controlling a single map into a predetermined trajectory. We analyze the reaction of the network to such a control. Specifically we show that a line of one-dimensional logistic maps that are unidirectionally coupled can be controlled from the first oscillator whereas a ring of diffusively coupled maps cannot be controlled for more than 5 maps. We show that rings with more elements can be controlled if every third map is controlled. The dependence of unidirectionally coupled maps on noise is studied. The noise level leads to a finite synchronization lengths for which maps can be controlled by a single location. A two-dimensional lattice is also studied. PMID- 15836277 TI - Dynamics of neuron populations in noisy environments. AB - In this paper different topologies of populations of FitzHugh-Nagumo neurons have been introduced in order to investigate the role played by the noise in the network. Each neuron is subjected to an independent source of noise. In these conditions the behavior of the population depends on the connection among the elements. By analyzing several kinds of topology (ranging from regular to random) different behaviors have been observed. Several topologies behave in an optimal way with respect to the range of noise level leading to an improvement in the stimulus response coherence, while others with respect to the maximum values of the performance index. However, the best results in terms of both the suitable noise level and high stimulus response coherence have been obtained when a diversity in neuron characteristic parameters has been introduced and the neurons have been connected in a small-world topology. PMID- 15836278 TI - Introduction: The Fermi-Pasta-Ulam problem--the first fifty years. PMID- 15836279 TI - Fermi-Pasta-Ulam, solitons and the fabric of nonlinear and computational science: history, synergetics, and visiometrics. AB - This paper is mostly a history of the early years of nonlinear and computational physics and mathematics. I trace how the counterintuitive result of near recurrence to an initial condition in the first scientific digital computer simulation led to the discovery of the soliton in a later computer simulation. The 1955 report by Fermi, Pasta, and Ulam (FPU) described their simulation of a one-dimensional nonlinear lattice which did not show energy equipartition. The 1965 paper by Zabusky and Kruskalshowed that the Korteweg-de Vries (KdV) nonlinear partial differential equation, a long wavelength model of the alpha lattice (or cubic nonlinearity), derived by Kruskal, gave quantitatively the same results obtained by FPU. In 1967, Zabusky and Deem showed that a localized short wavelength initial excitation (then called an "optical" and now a "zone-boundary mode" excitation ) of the alpha-lattice revealed "n-curve" coherent states. If the initial amplitude was sufficiently large energy equipartition followed in a short time. The work of Kruskal and Miura (KM), Gardner and Greene (GG), and myself led to the appreciation of the infinity of denumerable invariants (conservation laws) for Hamiltonian systems and to a procedure by GGKM in 1967 for solving KdV exactly. The nonlinear science field exponentiated in diversity of linkages (as described in Appendix A). Included were pure and applied mathematics and all branches of basic and applied physics, including the first nonhydrodynamic application to optical solitons, as described in a brief essay (Appendix B) by Hasegawa. The growth was also manifest in the number of meetings held and institutes founded, as described briefly in Appendix D. Physicists and mathematicians in Japan, USA, and USSR (in the latter two, people associated with plasma physics) contributed to the diversification of the nonlinear paradigm which continues worldwide to the present. The last part of the paper (and Appendix C) discuss visiometrics: the visualization and quantification of simulation data, e.g., projection to lower dimensions, to facilitate understanding of nonlinear phenomena for modeling and prediction (or design). Finally, I present some recent developments that are linked to my early work by: Dritschel (vortex dynamics via contour dynamics/surgery in two and three dimensions); Friedland (pattern formation by synchronization in Hamiltonian nonlinear wave, vortex, plasma, systems, etc.); and the author ("n-curve" states and energy equipartition in a FPU lattice). PMID- 15836280 TI - Long way from the FPU-problem to chaos. AB - This paper provides some historical comments on the study of the Fermi, Pasta, and Ulam (FPU) paper and its influence on the development of the theory of chaos. We also discuss some problems raised in the FPU paper and the links of these problems to such contemporary notions in chaos theory as ergodicity, mixing, recurrences, pseudochaos, kinetics, intermittency, etc. PMID- 15836281 TI - The Fermi-Pasta-Ulam problem: fifty years of progress. AB - A brief review of the Fermi-Pasta-Ulam (FPU) paradox is given, together with its suggested resolutions and its relation to other physical problems. We focus on the ideas and concepts that have become the core of modern nonlinear mechanics, in their historical perspective. Starting from the first numerical results of FPU, both theoretical and numerical findings are discussed in close connection with the problems of ergodicity, integrability, chaos and stability of motion. New directions related to the Bose-Einstein condensation and quantum systems of interacting Bose-particles are also considered. PMID- 15836282 TI - The Fermi-Pasta-Ulam problem as a challenge for the foundations of physics. AB - The Fermi-Pasta-Ulam (FPU) problem is discussed in connection with its physical relevance, and it is shown how apparently there exist only two possibilities: either the FPU problem is just a curiosity, or it has a fundamental role for the foundations of physics, casting a new light on the relations between classical and quantum mechanics. To this end, a short review is given of the main conceptual proposals that have been advanced. Particular emphasis is given to the perspective of a metaequilibrium scenario, which appears to be the only possible one for the FPU paradox to survive in the physically relevant case of infinitely many particles. PMID- 15836283 TI - Weak and strong chaos in Fermi-Pasta-Ulam models and beyond. AB - We briefly review some of the most relevant results that our group obtained in the past, while investigating the dynamics of the Fermi-Pasta-Ulam (FPU) models. The first result is the numerical evidence of the existence of two different kinds of transitions in the dynamics of the FPU models: (i) A stochasticity threshold (ST), characterized by a value of the energy per degree of freedom below which the overwhelming majority of the phase space trajectories are regular (vanishing Lyapunov exponents). It tends to vanish as the number N of degrees of freedom is increased. (ii) A strong stochasticity threshold (SST), characterized by a value of the energy per degree of freedom at which a crossover appears between two different power laws of the energy dependence of the largest Lyapunov exponent, which phenomenologically corresponds to the transition between weak and strong chaotic regimes. It is stable with N. The second result is the development of a Riemannian geometric theory to explain the origin of Hamiltonian chaos. Starting this theory has been motivated by the inadequacy of the approach based on homoclinic intersections to explain the origin of chaos in systems of arbitrarily large N, or arbitrarily far from quasi-integrability, or displaying a transition between weak and strong chaos. Finally, the third result stems from the search for the transition between weak and strong chaos in systems other than FPU. Actually, we found that a very sharp SST appears as the dynamical counterpart of a thermodynamic phase transition, which in turn has led, in the light of the Riemannian theory of chaos, to the development of a topological theory of phase transitions. PMID- 15836284 TI - Korteweg-de Vries equation and energy sharing in Fermi-Pasta-Ulam. AB - We address the problem of equipartition in a long Fermi-Pasta-Ulam (FPU) chain. After giving a precise relation between FPU and Korteweg-de Vries we use the latter equation to show that, corresponding to initial data a la Fermi, the time average of the energy on the kth mode decreases exponentially with kN. The result persists in the thermodynamic limit. PMID- 15836285 TI - Time scale for energy equipartition in a two-dimensional FPU model. AB - The FPU problem, i.e., the problem of energy equipartition among normal modes in a weakly nonlinear lattice, is here studied in dimension two, more precisely in a model with triangular cell and nearest-neighbors Lennard-Jones interaction. The number n of degrees of freedom ranges from 182 to 6338. Energy is initially equidistributed among a small number n(0) of low frequency modes, with n(0) proportional to n. We study numerically the time evolution of the so-called spectral entropy and the related "effective number" n(eff) of degrees of freedom involved in the dynamics; in this (rather typical) way we can estimate, for each n and each specific energy (energy per degree of freedom) epsilon, the time scale T(n)(epsilon) for energy equipartition. Numerical results indicate that in the thermodynamic limit the equipartition times are short: more precisely, for large n at fixed epsilon we find a limit curve T(infinity)(epsilon), and T(infinity) grows only as epsilon(-1) for small epsilon. Larger equipartition times are obtained by lowering epsilon, at fixed n, below a crossover value epsilon(c)(n). However, epsilon(c) appears to vanish by increasing n (faster than 1n), and the total energy E=nepsilon, rather than epsilon, appears to be the relevant variable when n is large and epsilon